PMID- 8702505 TI - Platelet endothelial cell adhesion molecule-1 (PECAM-1) homophilic adhesion is mediated by immunoglobulin-like domains 1 and 2 and depends on the cytoplasmic domain and the level of surface expression. AB - PECAM-1/CD31 is vascular cell adhesion and signaling molecule of the Ig superfamily that plays a role in neutrophil recruitment at inflammatory sites and may be involved the release of leukocytes from the bone marrow and in cardiovascular development. The interactions of PECAM-1 with its ligands are complex in that it is able to bind both with itself (homophilic adhesion) or with non-PECAM-1 ligands (heterophilic adhesion). Although the factors that regulate ligand binding are not fully understood, these interactions are regulated in part by its large cytoplasmic domain, a region of 118 amino acids encoded by 8 exons of its gene (exons 9-16). The purpose of this work was to better define the mechanisms of PECAM-1-dependent homophilic adhesion by analyzing the binding interactions of L-cells expressing full-length and selectively mutated forms of human, murine, and human/murine chimeric PECAM-1 molecules in an established aggregation assay. These studies demonstrate that 1) the minimal length of the cytoplasmic domain required for cellular aggregation is represented within the sequences encoded by exons 9 and 10, 2) removal or addition of the sequences encoded by exon 14 from the cytoplasmic domain can determine whether the mechanism of aggregation is a heterophilic calcium-dependent process or a homophilic calcium-independent process, 3) high levels of surface expression of PECAM-1 on the cell surface change the mechanism of aggregation from heterophilic to homophilic, and 4) PECAM-1-dependent homophilic binding appears to involve the direct interaction of only the first two extracellular Ig-like domains. These data suggest that PECAM-1-ligand interactions can be regulated through multiple pathways including alterations of the cytoplasmic domain and the level of surface expression. PMID- 8702506 TI - Enzymatic and biochemical probes of residues external to the translocation pathway of UhpT, the sugar phosphate carrier of Escherichia coli. AB - Part of the substrate translocation pathway through UhpT, the Escherichia coli sugar phosphate carrier, has been assigned to a transmembrane helix extending between residues 260 and 282. To set limits on the external portion of the pathway, we identified nearby residues fully exposed to the periplasm. In one case, we used Western blots to evaluate cleavage by extracellular trypsin. The protease cleaved UhpT variants retaining lysine 294, but not those lacking lysine 294, indicating that trypsin acts at a single extracellular site, lysine 294. In other work we labeled single-cysteine variants with 3-(N maleimidylpropionyl)biocytin and scored accessibility to extracellular streptavidin by shifts of SDS-polyacrylamide gel electrophoresis mobility. Positions 283 and 284 were fully exposed to the periplasm, since the modified residue was bound by streptavidin in the native protein; by contrast, although the biotin-linked probe modified position 276, streptavidin decoration was not achieved without protein denaturation. We conclude that a 12-residue stretch(283 294) of UhpT is sufficiently exposed to be accessible to large probes (trypsin, streptavidin), while position 276 and more proximal residues are more deeply buried or otherwise shielded from the external phase. PMID- 8702507 TI - Egr-1 activates basic fibroblast growth factor transcription. Mechanistic implications for astrocyte proliferation. AB - The mechanisms controlling the proliferation of astrocytes are of great interest but are not well defined. We have previously shown that the endogenous neuropeptides, endothelin-3 (ET-3), and atrial natriuretic peptide (ANP), modulate the proliferation of astrocytes through positively and negatively regulating the transcription of the immediate-early gene egr-1 which transactivates basic fibroblast growth factor (bFGF) by unknown mechanisms. In these studies, we determined the involvement of MAP kinase (Erk) activation by ET 3 in the transcription of egr-1, and the molecular determinants by which Egr-1 transactivates bFGF. Transfection of astrocytes with a mitogen-activated protein (MAP) kinase (MAPK) expression vector increased the transcription of a cotransfected egr-chloramphenicol acetyltransferase (CAT) construct 3-fold. This induction was totally abolished by a dominant negative MAPK mutant. A 3-fold induction of egr-CAT expression by ET-3 was significantly reduced by treatment with ANP, or a cotransfected dominant negative MAPK plasmid. Using mobility shift assays, we showed that ET-3 induced the expression of Egr-1 protein which bound specifically to several early growth-related protein (Egr-1) binding sites on the bFGF promoter, and that this effect was significantly reversed by treatment with ANP. We also found that the Sp1 transcriptional factor was bound at these same sites, but was not stimulated by ET-3. Deletion experiments indicated that only the site at -160 bp of the bFGF promoter was significant for bFGF transactivation by Egr-1. We conclude that the astrocyte mitogen, ET-3, stimulates egr-1 transcription through a MAP kinase (Erk) related mechanism, and that Egr-1 transactivates bFGF through a specific noncanonical, Egr-1 site on the promoter. ANP inhibits each of these steps, providing a pathway for its anti-proliferative action. PMID- 8702508 TI - G protein gamma subunits with altered prenylation sequences are properly modified when expressed in Sf9 cells. AB - The gamma subunits of heterotrimeric G proteins undergo post-translational prenylation and carboxylmethylation after formation of the betagamma dimer, modifications that are essential for alpha-betagamma, betagamma-receptor, and betagamma-effector interactions. We have determined the specific prenyl group present on the beta1gamma1, beta1gamma2, and beta1gamma3 dimers purified from baculovirus-infected Sf9 cells by specific binding to G protein alpha subunits immobilized on agarose. These recombinant dimers undergo the same post translational modifications determined for gamma1 and gamma2 isolated from mammalian tissues. Furthermore, infection of Sf9 cells with a recombinant baculovirus encoding an alteration of the gamma1 CaaX sequence (gamma1-S74L) resulted in geranylgeranylation of the resulting gamma1 subunit, and alteration of the gamma2 CaaX sequence to CAIS (gamma2-L71S) resulted in farnesylation. Both of these altered gamma subunits were able to associate stably with beta1, and the resulting betagamma dimer bound tightly to alpha-agarose and eluted specifically with aluminum fluoride. These results indicate that Sf9 insect cells properly process the CaaX motif in G protein gamma subunits and are a useful model system to study the role of prenylation in the protein-protein interactions in which the betagamma subunits participate. PMID- 8702509 TI - Role of the prenyl group on the G protein gamma subunit in coupling trimeric G proteins to A1 adenosine receptors. AB - The coupling of receptors to heterotrimeric G proteins is determined by interactions between the receptor and the G protein alpha subunits and by the composition of the betagamma dimers. To determine the role of the gamma subunit prenyl modification in this interaction, the CaaX motifs in the gamma1 and gamma2 subunits were altered to direct modification with different prenyl groups, recombinant betagamma dimers expressed in the baculovirus/Sf9 insect cell system, and the dimers purified. The activity of the betagamma dimers was compared in two assays: formation of the high affinity agonist binding conformation of the A1 adenosine receptor and receptor-catalyzed exchange of GDP for GTP on the alpha subunit. The beta1gamma1 dimer (modified with farnesyl) was significantly less effective than beta1gamma2 (modified with geranylgeranyl) in either assay. The beta1gamma1-S74L dimer (modified with geranylgeranyl) was nearly as effective as beta1gamma2 in either assay. The beta1gamma2-L71S dimer (modified with farnesyl) was significantly less active than beta1gamma2. Using 125I-labeled betagamma subunits, it was determined that native and altered betagamma dimers reconstituted equally well into Sf9 membranes containing A1 adenosine receptors. These data suggest that the prenyl group on the gamma subunit is an important determinant of the interaction between receptors and G protein gamma subunits. PMID- 8702510 TI - Mechanism of nitric oxide release from S-nitrosothiols. AB - S-Nitrosothiols have many biological activities and have been suggested to be intermediates in signal transduction. The mechanism and products of S nitrosothiol decomposition are of great significance to the understanding of nitric oxide (.NO) biochemistry. S-Nitrosothiols are stable compounds at 37 degrees C and pH 7.4 in the presence of transition metal ion chelators. The presence of trace transition metal ions (present in all buffers) stimulates the catalytic breakdown of S-nitrosothiols to .NO and disulfide. Thiyl radicals are not formed as intermediates in this process. Photolysis of S-nitrosothiols results in the formation of .NO and disulfide via the intermediacy of thiyl radicals. Reduced metal ion (e.g. Cu+) decomposes S-nitrosothiols more rapidly than oxidized metal ion (e.g. Cu2+) indicating that reducing agents such as glutathione and ascorbate can stimulate decomposition of S-nitrosothiol by chemical reduction of contaminating transition metal ions. Transnitrosation can also stimulate S-nitrosothiol decomposition if the product S-nitrosothiol is more susceptible to transition metal ion-catalyzed decomposition than the parent S nitrosothiol. Equilibrium constants for the transnitrosation reactions of reduced glutathione, either with S-nitroso-N-acetyl-dl-penicillamine or with S-nitroso-L cysteine indicate that S-nitrosoglutathione formation is favored. The biological relevance of S-nitrosothiol decomposition is discussed. PMID- 8702511 TI - Role of the proposed serpin-enzyme complex receptor recognition site in binding and internalization of thrombin-heparin cofactor II complexes by hepatocytes. AB - Several serpin-enzyme complexes bind to a receptor on hepatocytes that mediates their endocytosis and lysosomal degradation. Joslin et al. (Joslin, G., Fallon, R. J., Bullock, J., Adams, S. P., and Perlmutter, D. H.(1991) J. Biol. Chem. 266, 11282-11288) proposed that a sequence near the C-terminal end of the serpin (e.g. FVFLM in alpha1-antitrypsin) binds to the serpin-enzyme complex receptor (SEC receptor). In experiments with synthetic peptides, they found that substitution of alanine at the fourth or fifth position in this sequence reduced the affinity of peptide binding to Hep G2 cells. To test the hypothesis that the corresponding sequence in heparin cofactor II (HCII), FLFLI (residues 456-460), mediates binding and uptake of the thrombin-HCII complex by Hep G2 cells, we constructed five recombinant HCII variants, F456A, L457A, F458A, L459A, and I460A. At 4 degrees C, the 125I-thrombin-HCII(native) complex bound reversibly to 0.6-2.6 x 10(5) sites per Hep G2 cell with a Kd of 19-32 nM. Binding was inhibited by excess unlabeled thrombin-HCII(native), thrombin-antithrombin, or elastase-alpha1 antitrypsin, but not by free HCII or thrombin, which is consistent with the reported specificity of the SEC receptor. However, complexes of thrombin with each of the HCII variants inhibited binding as effectively as the complex with native HCII. Competitive binding experiments with various concentrations of unlabeled thrombin-HCII(native) or thrombin-HCII(I460A) indicated that these complexes bind to Hep G2 cells with equal affinity. At 37 degrees C, complexes of 125I-thrombin with each of the five HCII variants were internalized and degraded at the same rate as the complex with native HCII. Our data suggest that the pentapeptide FLFLI in HCII is not involved in binding, internalization, and degradation of thrombin-HCII complexes by Hep G2 cells. PMID- 8702512 TI - Critical residues of integrin alphaIIb subunit for binding of alphaIIbbeta3 (glycoprotein IIb-IIIa) to fibrinogen and ligand-mimetic antibodies (PAC-1, OP G2, and LJ-CP3). AB - Integrin alphaIIbbeta3 plays a critical role in platelet aggregation through its interaction with fibrinogen. Elucidation of the mechanisms of alphaIIbbeta3 fibrinogen interaction is critical to understanding hemostasis and thrombosis. Here we report that mutations of Gly-184, Tyr-189, Tyr-190, Phe-191, and Gly-193 within the predicted turn structure of the third amino-terminal repeat of alphaIIb significantly block binding of alphaIIbbeta3 to soluble fibrinogen. These mutations also block binding of alphaIIbbeta3 to ligand-mimetic monoclonal antibodies PAC-1, OP-G2, LJ-CP3, which have an RGD-related RYD sequence in their antigen-binding sites. These mutations do not significantly affect the expression of alphaIIbbeta3, in contrast to most of the natural alphaIIb mutations occurring in Glanzmann's thrombasthenic patients. The data suggest that these residues are critically involved in alphaIIbbeta3-ligand interactions. PMID- 8702513 TI - Rapid flux in transforming growth factor-beta receptors on bone cells. AB - The proportion of transforming growth factor-beta (TGF-beta) binding among conventional membrane receptors on bone cells can vary with hormone or growth factor treatment or with the state of osteoblast-like activity and appears to determine the nature of its biological effects. Therefore, functional TGF-beta receptor stability could be an important aspect of regulation. Suppression of protein synthesis reduced TGF-beta binding to types I and II receptors with t1/2 of 2 h and to betaglycan with t1/2 of 6 h. In contrast, suppression of mRNA transcription reduced TGF-beta binding at least 3-fold more slowly at each receptor site. Preexposure to TGF-beta decreased its binding at all three sites within 4 h in osteoblast-enriched cultures. This effect was transient with lower TGF-beta concentrations, where the receptor profile was nearly fully restored within 24-48 h. In contrast, less differentiated bone cells were less sensitive to ligand-dependent receptor down-regulation. Agents that alter protein kinase and phosphatase activity also modified the TGF-beta binding profile in specific ways. Together, these results indicate that cell surface TGF-beta receptors turn over rapidly by ligand-independent and ligand-dependent mechanisms, demonstrate that the binding capacity of TGF-beta receptors is less stable than their mRNAs, and that functional receptor levels may be determined in part by post transcriptional events. PMID- 8702514 TI - Feedback regulation of hepatic 7alpha-hydroxylase expression by bile salts in the hamster. AB - Hepatic 7alpha-hydroxylase activity appears to be regulated at the transcriptional level by the quantity of bile salts fluxing through the enterohepatic circulation. Whether bile salts directly suppress 7alpha hydroxylase expression at the level of the hepatocyte or do so indirectly by promoting the release or absorption of an intestinal factor has not been resolved. We have investigated the ability of primary bile salts to suppress hepatic 7alpha-hydroxylase expression in bile-diverted hamsters. Biliary diversion was accompanied by derepression of both hepatic 7alpha-hydroxylase activity (4-5-fold) and bile salt secretion (approximately 3-fold). Derepression of hepatic 7alpha-hydroxylase expression could be prevented by several interventions that increase the availability of bile salts within the hepatocyte including 1) overexpression of an exogenous 7alpha-hydroxylase gene by adenovirus mediated gene transfer, 2) obstruction of the common bile duct, and 3) intravenous infusions of taurocholate. In contrast, none of these interventions prevented derepression of hepatic cholesterol synthesis or significantly down regulated hepatic low density lipoprotein receptor expression over the relatively short time course (24 h) of these studies. Together, these data indicate that primary bile salts contribute to the regulation of bile salt synthesis through feedback repression of 7alpha-hydroxylase expression at the level of the hepatocyte. PMID- 8702515 TI - Characterization of the mononickel metallocenter in H134A mutant urease. AB - A mutant form of Klebsiella aerogenes urease possessing Ala instead of His at position 134 (H134A) is inactive and binds approximately half the normal complement of nickel (Park, I.-S., and Hausinger, R. P.(1993) Protein Sci. 2, 1034-1041). The crystal structure of the H134A protein was obtained at 2.0-A resolution, and it confirms that only Ni-1 of the two nickel ions found in the native enzyme is present. In contrast to the pseudotetrahedral geometry observed for Ni-1 in native urease (where it is liganded by His-246, His-272, one oxygen atom of carbamylated Lys-217, and a water molecule at partial occupancy), the mononickel metallocenter in the H134A protein was found to possess octahedral geometry and was coordinated by the above protein ligands plus three water molecules. The nickel site of H134A urease was probed by UV-visible, variable temperature magnetic circular dichroism, and x-ray absorption spectroscopies. The spectroscopic data are consistent with the presence of Ni(II) in octahedral geometry coordinated by two histidylimidazoles and additional oxygen and/or nitrogen donors. These data underscore the requirement of Ni-2 for formation of active urease and demonstrate the important role of Ni-2 in establishing the proper Ni-1 coordination geometry. PMID- 8702516 TI - Oxidation of sulfhydryl groups of ribonuclease inhibitor in epithelial cells is sufficient for its intracellular degradation. AB - Ribonuclease inhibitor (RI) is a cytoplasmic protein (50 kDa) that inhibits a variety of pancreatic type RNases. The porcine inhibitor contains 30 cysteine residues, all of which occur in the reduced state. It is well known that in vitro modification of the thiol groups inactivates the protein and greatly increases its susceptibility to proteolysis. Here we show that oxidation of thiol groups in RI can also occur within the cell. Induction of an oxidative insult in cultured LLC-PK1 cells, either with a general oxidant, H2O2, or with a thiol-specific oxidant, diamide, led to the loss of RI activity. By using specific antibodies it was demonstrated that the decrease correlated with a decline in the amount of RI protein in the cells. Furthermore, analysis of RI mRNA levels and half-life of the protein excluded inhibition of the synthesis of RI as the cause of its depletion. The results indicate that oxidation of thiol groups in RI is sufficient to cause its rapid inactivation and disappearance from the cell. Most likely this results from intracellular degradation of the protein. PMID- 8702517 TI - Expression cloning of lsc, a novel oncogene with structural similarities to the Dbl family of guanine nucleotide exchange factors. AB - In a screen for genes with oncogenic potential expressed by the murine B6SUtA1 myeloid progenitor cell line, we isolated a 2. 5-kilobase pair cDNA whose expression causes strong morphological transformation and deregulated proliferation of NIH 3T3 cells. The transforming cDNA encodes a truncated protein (designated Lsc) with a region of sequence similarity to the product of the lbc oncogene. This region includes the tandem Dbl homology and pleckstrin homology domains that are hallmarks of the Dbl-like proteins, a family of presumptive or demonstrated guanine nucleotide exchange factors that act on Rho family GTPases. Lsc requires intact Dbl homology and pleckstrin homology domains for its oncogenic activity. The transforming activity of Lsc in NIH 3T3 cells is reduced by cotransfection with p190 (a GTPase activating protein for Rho family GTPases) and the Rho family dominant-negative mutants RhoA(19N), CDC42(17N), and Rac1(17N). These results indicate a role for the Rho family of GTPases in mediating the transforming activity of Lsc and are consistent with the exchange specificities that have been attributed to Dbl family members. The lsc gene is expressed in a variety of tissues and is particularly abundant in hemopoietic tissues (thymus, spleen, and bone marrow). Lsc is a member of a growing family of proteins that may function as activators of Rho family GTPases in a developmental or tissue-specific manner. PMID- 8702518 TI - Prostaglandin E2 stimulates a Ca2+-dependent K+ channel in human erythrocytes and alters cell volume and filterability. AB - To understand the mechanism by which human red blood cells (RBCs) contribute to hemostasis and thrombosis, we have examined the effects of metabolites released by activated platelets on intact RBCs. Prostaglandin E2 (PGE2), a signal molecule produced by activated platelets, was observed to lower the filterability of human erythrocytes by approximately 30% at 10(-10) M. PGE2 also caused a reduction in mean cell volume of approximately 10%. The shrinkage of red cells after PGE2 treatment was confirmed by documenting a decrease in osmotic fragility and an increase in cell density following exposure to the hormone. Careful analysis, however, revealed that only approximately 15% of the erythrocytes responded to stimulation with PGE2. Examination of the cause of cell shrinkage showed that induction of a PGE2-stimulated K+ efflux pathway leading to rapid loss of cellular K+ was responsible. The PGE2-stimulated K+ loss was also observed to be Ca2+-dependent, suggesting the possible involvement of the Gardos channel. Gardos channel participation was supported by the observation that two Gardos channel inhibitors, charybdotoxin and clotrimazole, independently blocked the PGE2 stimulated K+ efflux. Further evidence for Gardos channel activation came from experiments aimed at characterizing the efflux pathway followed by the obligatory counterion. Thus, K+ efflux was readily stimulated even when NO3- was substituted for Cl-, suggesting that neither KCl cotransport nor Na/K/2Cl cotransport plays a prominent role in the PGE2-induced cell shrinkage. Further, the anion transporter band 3 was implicated as the counterion efflux route, since DIDS inhibited the PGE2-stimulated cell volume change without blocking the change in membrane potential. Taken together, we propose that release of PGE2 by activated platelets constitutes part of a mechanism by which activated platelets may recruit adjacent erythrocytes to assist in clot formation. PMID- 8702519 TI - Cloning of the sodium-dependent, broad-scope, neutral amino acid transporter Bo from a human placental choriocarcinoma cell line. AB - We have isolated a cDNA from a human placental choriocarcinoma cell cDNA library which, when expressed in HeLa cells, induces a Na+-dependent amino acid transport system with preference for zwitterionic amino acids. Anionic amino acids, cationic amino acids, imino acids, and N-methylated amino acids are excluded by this system. These characteristics are identical to those described for the amino acid transporter Bo. When expressed in Xenopus laevis oocytes that do not have detectable endogenous activity of the amino acid transporter Bo, the cloned transporter increases alanine transport in the oocytes severalfold and induces alanine-evoked inward currents in the presence of Na+. The cDNA codes for a polypeptide containing 541 amino acids with 10 putative transmembrane domains. Amino acid sequence homology predicts this transporter (hATBo) to be a member of a superfamily consisting of the glutamate transporters, the neutral amino acid transport system ASCT, and the insulin-activable neutral/anionic amino acid transporter. Chromosomal assignment studies with somatic cell hybrid analysis and fluorescent in situ hybridization have located the ATBo gene to human chromosome 19q13.3. PMID- 8702520 TI - Expression of human glucocorticoid receptor gene and interaction of nuclear proteins with the transcriptional control element. AB - We have identified sequences responsible for the expression of the human glucocorticoid receptor gene (GR gene) using a set of 5' promoter deletion mutants in HeLa, human placenta, and human breast tumor (MCF-7) cells. The chimeric gene construct -892 5'-GAAGTGACACACTTC3' -878-CAT was sufficient for high level of expression in HeLa and placenta cells in culture. Deletion of palindromic sequences decreased levels of GR expression in these cells. By oligonucleotide-affinity chromatography with the palindromic glucocorticoid receptor enhancing factor-binding element (GREFE), we have isolated from human placenta nuclear extract two novel proteins glucocorticoid receptor enhancing factors 1 and 2 (GREF1 and GREF2), with apparent molecular masses of 80 and 62 kDa, respectively. These proteins, similar to the DNA-binding autoantigen Ku are, like Ku, heterodimers of polypeptide subunits p80 and p62, immunologically related to factors binding to proximal sequence element 1 in the promoter of small nuclear RNA (PSE1) and transferrin receptor enhancing factors. Both Ku80 and Ku70 polypeptides were present in high concentrations in human placenta and HeLa cells. In MCF-7 cells, however, only a high level of p62 was detected. While cotransfection of pcDNA-Ku80 with pHGR(-892 to -878)-CAT potentiated the expression of CAT, introduction of pcDNA-Ku70 did not affect the expression of CAT in transfected MCF-7 cells. UV cross-linking analysis showed that only GREF1 contacted DNA directly. Supershift assays with monoclonal antibodies Ab 111 (Ku80) or Ab N3H10 (Ku70) showed a direct interaction of GREF1 and GREF2 heterodimers with the palindrome. Partial peptide fingerprinting of GREF1 and GREF2 using alpha-chymotrypsin and immunoblotting with Ab 111 and Ab N3H10 confirmed their identities as Ku80 and Ku70, respectively. PMID- 8702521 TI - Molecular oxygen modulates cytochrome c oxidase function. AB - This study sought to determine whether molecular oxygen interacts with cytochrome c oxidase to modify its catalytic activity. Such an interaction could explain the observation that mitochondria incubated under low O2 concentrations exhibit a reversible suppression of State 3 respiration. Oxidized bovine heart cytochrome c oxidase was incubated in oxygen concentrations of <50 microM for 4 h. The enzyme exhibited a reversible decrease in Vmax after incubation, compared with control enzyme incubated at higher oxygen concentrations. This change was accompanied by a small increase in the apparent Km of the enzyme for both cytochrome c and oxygen, although the optical absorption spectra of oxidized, cycling, or reduced enzyme were not affected. Spectroscopy studies after 4 h of incubation revealed that heme a3 was 33% reduced during cycling at [O2] = 25 microM whereas enzyme at [O2] = 135 microM was only 18% reduced, suggesting that the site of inhibition occurred at the electron transfer step between heme a3 and O2. These results provide a mechanistic explanation for the observation that intact cells or mitochondria exhibit a reversible inhibition of respiration during prolonged exposure to [O2] <25 mM, by demonstrating that the catalytic activity of cytochrome c oxidase function is similarly inhibited, possibly through an allosteric effect of molecular O2 on the enzyme. PMID- 8702522 TI - A new cyclic AMP-independent, Gs-mediated stimulatory mechanism via the adenosine A2a receptor in the intact cardiac cell. AB - The objectives of this study were to investigate the mechanism underlying the adenosine A2a receptor (A2aR)-mediated positive inotropic response and to define its contractile function using chick embryo ventricular cells as a model. Activation of the A2aR caused a marked stimulation of calcium entry and cell contractility, which were blocked by verapamil or nifedipine. The effects elicited by maximal concentrations of the A2aR agonist 2-[4-(2 carboxyethyl)phenylethylamino]-5'-N-ethylcarboxamidoadenos ine and the beta adrenergic agonist isoproterenol were additive, indicating that the two receptors do not share a common stimulatory mechanism. The cAMP antagonist (Rp)-adenosine cyclic 3':5'-monophosphorothioate was ineffective in inhibiting the A2aR-mediated stimulation of contractility or the L-type calcium channel, while it completely abolished the isoproterenol effects. Activation of the A2aR had no effect on Na+/Ca2+ exchange or inositol 1,4,5-trisphosphate accumulation. Blocking of the A2aR resulted in unopposed A1 receptor-mediated inhibitory effects and led to an inhibition of basal contractility and an enhanced anti-adrenergic effect by A1 agonist. The adenosine A2a receptor mediates a new cyclic AMP-independent mechanism and a new contractile function in the cardiac cell. PMID- 8702523 TI - Interactions between microsomal triglyceride transfer protein and apolipoprotein B within the endoplasmic reticulum in a heterologous expression system. AB - When apolipoprotein B (apoB) is expressed in heterologous cells, it is not secreted but retained and degraded within the endoplasmic reticulum (ER). We have previously characterized carboxyl-terminal truncated forms of apoB expressed in COS cells and have shown that these proteins were readily synthesized but retained within the ER and degraded, if the size of the truncated protein was larger than apoB 29. Below this size, the smaller the size of the apoB truncates, the greater the extent of secretion, although >50% of these smaller proteins were also degraded within the ER. In the present study, we demonstrate that this secretory defect can be overcome by coexpression with microsomal triglyceride transfer protein (MTP); moreover, this complementation is inversely related to the size of apoB. Secretion of apoBs larger than B29 required the coexpression of MTP and, in the presence of MTP, was oleate-responsive. MTP, in the presence or absence of oleate supplementation, had little or no effect on the secretion of the shorter truncates. We discovered, however, that MTP was physically associated with all forms of apoB intracellularly (B13-B41). The association of MTP with apoB 41 was stable to high salt washing, as well as to low pH, suggesting that these interactions may be hydrophobic in nature. In addition to the interaction with MTP, apoB was also found to be associated with calnexin, confirming previous studies, and with proteins bearing the KDEL retention signal. However, studies on overexpression of human calnexin and tunicamycin inhibition of glycosylation showed that interaction with calnexin was not necessary for the formation or secretion of apoB 41-containing lipoproteins; moreover, in the presence of MTP, the association of calnexin with apoB 41 was transient or absent. These data suggest that for apoB to attain a folded state sufficient to escape the quality control of the ER, it needs to obtain neutral lipid (supplied by MTP), as well as its ability to keep it packaged as a rudimentary lipoprotein, dependent on its size being larger than B29. PMID- 8702524 TI - Molecular shape, dissociation, and oxygen binding of the dodecamer subunit of Lumbricus terrestris hemoglobin. AB - Small angle x-ray scattering of the 213-kDa dodecamer of Lumbricus terrestris Hb yielded radius of gyration = 3.74 +/- 0.01 nm, maximum diameter = 10.59 +/- 0.01 nm, and volume = 255 +/- 10 nm3, with no difference between the oxy and deoxy states. Sedimentation velocity studies indicate the dodecamer to have a spherical shape and concentration- and Ca2+-dependent equilibria with its constituent subunits, the disulfide-bonded trimer of chains a-c and chain d. Equilibrium sedimentation data were fitted best with a trimer-dodecamer model, ln K4 = 7 (association K in liters3/g3) at 1 degrees C and 4 at 25 degrees C, providing DeltaH = -20 kcal/mol and DeltaS = 4.4 eu/mol. Oxydodecamer dissociation at pH 8.0, in urea, GdmCl, heteropolytungstate K8[SiW11O39] and of metdodecamer at pH 7, was followed by gel filtration. Elution profiles were fitted with exponentially modified gaussians to represent the three peaks. Two exponentials were necessary to fit all the dissociations except in [SiW11O39]-8. Equilibrium oxygen binding measurements at pH 6.5-8. 5, provided P50 = 8.5, 11.5-11.9 and 11.9-13.5 torr, and n50 = 5.2-9. 5, 3.2-4.9, and 1.8-2.7 for blood, Hb, and dodecamer, respectively, at pH 7.5, 25 degrees C. P50 was decreased 3- and 2-fold in approximately 100 mM Ca2+ and Mg2+, respectively, with concomitant but smaller increases in cooperativity. PMID- 8702525 TI - Autoregulation of the plasmid addiction operon of bacteriophage P1. AB - The P1 plasmid addiction operon increases the apparent stability of a plasmid that carries it by killing plasmid-free (cured) segregants. The operon consists of a gene encoding an endotoxin responsible for death on curing (doc), preceded by a gene encoding a relatively unstable antidote that can prevent host death (phd). When the copy number of the operon was increased, expression of a lacZ reporter fused to the promoter of the operon decreased, indicating that expression of the operon was stabilized by an autoregulatory circuit. Transcription of the lacZ reporter was repressed about 10-fold when phd, without doc, was expressed from an exogenous promoter. DNase I footprinting showed that Phd binds a perfect 10-base pair palindromic DNA sequence and, at higher concentrations, an adjacent, imperfect palindrome. The palindromic sites are located between the -10 region of the putative promoter and the start codon of phd. Electrophoretic mobility of DNA containing the promoter region was retarded in the presence of Phd and further retarded in the presence of Phd and Doc. When doc was co-expressed with phd, repression of the lacZ fusion was enhanced more than 100-fold. Thus, both products of the addiction operon participate in its autoregulation. PMID- 8702526 TI - Specific tryptophan substitution in catalytic sites of Escherichia coli F1-ATPase allows differentiation between bound substrate ATP and product ADP in steady state catalysis. AB - Tryptophan was specifically inserted as the residue immediately preceding the P loop sequence in F1-ATPase catalytic sites. The mutant enzyme (betaF148W) showed normal enzymatic characteristics. The fluorescence responses of beta-tryptophan 148 enabled us to differentiate between nucleoside di- and triphosphate bound in catalytic sites; MgADP quenched at 350 nm, whereas MgAMPPNP and MgADP.BeFx complex enhanced the fluorescence at 325 nm. With MgATP, both effects were seen simultaneously. This allowed analysis of bound catalytic site nucleotides directly under steady-state MgATP hydrolysis conditions. At mM concentration of MgATP (Vmax conditions) one of the three catalytic sites was filled with substrate MgATP and the other two sites were filled with product MgADP. A model for F1-ATPase steady-state turnover is presented that encompasses these findings. Given the structural similarity of the P-loop in nucleotide-binding proteins, this approach may prove widely useful. PMID- 8702527 TI - An insulin receptor mutant (Asp707 --> Ala), involved in leprechaunism, is processed and transported to the cell surface but unable to bind insulin. AB - We have identified a homozygous mutation near the carboxyl terminus of the insulin receptor (IR) alpha subunit from a leprechaun patient, changing Asp707 into Ala. Fibroblasts from this patient had no high affinity insulin binding sites. To examine the effect of the mutation on IR properties, the mutant IR was stably expressed in Chinese hamster ovary cells. Western blot analysis and metabolic labeling showed a normal processing of the mutant receptor to alpha and beta subunits. No increase in high affinity insulin binding sites was observed on Chinese hamster ovary cells expressing the mutant receptor, and also, affinity cross-linking of 125I-labeled insulin by disuccinimidyl suberate to these cells failed to label the mutant alpha subunit. Biotinylation of cell surface proteins by biotin succinimidyl ester resulted in efficient biotinylation of the mutant IR alpha and beta subunits, showing its presence on the cell surface. On solubilization of the mutant insulin receptor in Triton X-100-containing buffers, 125I-insulin was efficiently cross-linked to the receptor alpha subunit by disuccinimidyl suberate. These studies demonstrate that Ala707 IR is normally processed and transported to the cell surface and that the mutation distorts the insulin binding site. Detergent restores this site. This is an example of a naturally occurring mutation in the insulin receptor that affects insulin binding without affecting receptor transport and processing. This mutation points to a major contribution of the alpha subunit carboxyl terminus to insulin binding. PMID- 8702528 TI - Methylcobamide:coenzyme M methyltransferase isozymes from Methanosarcina barkeri. Physicochemical characterization, cloning, sequence analysis, and heterologous gene expression. AB - A comparative study was made on the physicochemical characteristics of two isozymes of methylcobamide:- coenzyme M methyltransferase (MT2). Both isozymes catalyzed S-methylation of 2-thioethanesulfonate (coenzyme M) and exhibited similar apparent Km values for coenzyme M of 35 microM (MT2-A) and 20 microM (MT2 M). Weak binding to methylcobalamin was indicated by the apparent Km of 14 mM for both isozymes. Cob(I)alamin was established as the major product of the reaction, demonstrating heterolytic cleavage of the methylcobamide carbon-cobalt bond. The isozymes were shown to be zinc-containing metalloproteins. Metal ion chelators strongly inhibited both isozymes. A variety of coenzyme M analogs were tested for activity and/or inhibition. One alternative substrate 3-mercaptopropionate was discovered, with apparent Km 9 mM (MT2-A) and 10 mM (MT2-M). The results suggested an active site geometry in which coenzyme M is bound both by S coordination to zinc, and electrostatic interaction of the sulfonate with a cationic group on the enzyme. Methanosarcina barkeri genes cmtA and cmtM encoding both isozymes were cloned and sequenced. Both genes encoded proteins with 339 amino acids and predicted molecular masses of 36-37 kDa. Active forms of both isozymes were expressed in Escherichia coli. A conserved segment with the potential for metal binding was found. The possibility of zinc involvement in catalysis of coenzyme M methylation is considered. PMID- 8702529 TI - Structures of the O-glycans on P-selectin glycoprotein ligand-1 from HL-60 cells. AB - P-selectin glycoprotein ligand-1 (PSGL-1) is a disulfide-bonded homodimeric mucin like glycoprotein on leukocytes that interacts with both P- and E-selectin. In this report we describe the structures of the Ser/Thr-linked O-glycans of PSGL-1 synthesized by HL-60 cells metabolically radiolabeled with 3H-sugar precursors. In control studies, the O-glycans on CD43 (leukosialin), a mucin-like glycoprotein also expressed by HL-60 cells, were analyzed and compared to those of PSGL-1. O-Glycans were released from Ser/Thr residues by mild base/borohydride treatment of purified glycoproteins, and glycan structures were determined by a combination of techniques. In contrast to expectations, PSGL-1 is not heavily fucosylated; a majority of the O-glycans are disialylated or neutral forms of the core-2 tetrasaccharide Galbeta1-->4GlcNAcbeta1-->6(Galbeta1-->3)GalNAcOH++ +. A minority of the O-glycans are alpha-1,3-fucosylated that occur as two major species containing the sialyl Lewis x antigen; one species is a disialylated, monofucosylated glycan, and the other is a monosialylated, trifucosylated glycan having a polylactosamine backbone. CD43 lacks the fucosylated glycans found on PSGL-1 and is enriched for the nonfucosylated, disialylated core-2 hexasaccharide. These results demonstrate that PSGL-1 contains unique fucosylated O-glycans that are predicted to be critical for high affinity interactions between PSGL-1 and selectins. PMID- 8702530 TI - EPR and Mossbauer spectroscopic studies on enoate reductase. AB - Enoate reductase (EC 1.3.1.31) is a protein isolated from Clostridium tyrobutyricum that contains iron, labile sulfide, FAD, and FMN. The enzyme reduces the alpha,beta carbon-carbon double bond of nonactivated 2-enoates and in a reversible way that of 2-enals at the expense of NADH or reduced methyl viologen. UV-visible and EPR potentiometric titrations detect a semiquinone species in redox intermediate states characterized by an isotropic EPR signal at g = 2.0 without contribution at 580 nm. EPR redox titration shows two widely spread mid-point redox potentials (-190 and -350 mV at pH 7. 0), and a nearly stoichiometric amount of this species is detected. The data suggest the semiquinone radical has an anionic nature. In the reduced form, the [Fe-S] moiety is characterized by a single rhombic EPR spectrum, observed in a wide range of temperatures (4. 2-60 K) with g values at 2.013, 1.943, and 1.860 (-180 mV at pH 7.0). The gmax value is low when compared with what has been reported for other iron-sulfur clusters. Mossbauer studies reveal the presence of a [4Fe-4S]+2/+1 center. One of the subcomponents of the spectrum shows an unusually large value of quadrupole splitting (ferrous character) in both the oxidized and reduced states. Substrate binding to the reduced enzyme induces subtle changes in the spectroscopic Mossbauer parameters. The Mossbauer data together with known kinetic information suggest the involvement of this iron-sulfur center in the enzyme mechanism. PMID- 8702531 TI - mRNA profiling of rat islet tumors reveals nkx 6.1 as a beta-cell-specific homeodomain transcription factor. AB - Development of a high capacity multiplex reverse transcriptase-polymerase chain reaction protocol has allowed us to screen lineage related rat islet tumors classified as alpha-, beta-, and delta-like as judged by their hormone profile for differential expression of more than 50 selected genes. We find that in addition to insulin the insulinoma express the normal beta-cell markers Pdx-1, IAPP, and Glut-2, and that these markers are absent from the glucagonoma: a reflection of the normal alpha-cell. Furthermore, this study suggests that the GLP-1, glucagon, GIP, IGF-1, and insulin receptors as well as E-cadherin, R cadherin, Id-1, and Id-2 are differentially expressed within the islet of Langerhans. Importantly, insulinoma-specific expression of the recently cloned homeodomain protein Nkx 6.1 predicted beta-cell-specific expression in the normal islet. Immunohistochemistry using antibodies raised against recombinant Nkx 6.1 did indeed localize Nkx 6.1 expression exclusively to the nuclei of normal islet beta-cells. Apart from pancreatic islets only the antral part of the stomach contained Nkx 6.1 mRNA. We conclude that multiplex reverse transcriptase polymerase chain reaction-based mRNA profiling is a powerful tool to identify differentially expressed genes within phenotypically related cells and propose that Nkx 6.1 is involved in specifying the unique characteristics of the beta cell. PMID- 8702532 TI - Suppression of syndecan-1 expression in endothelial cells by tumor necrosis factor-alpha. AB - Syndecan-1 is a cell surface proteoglycan that binds extracellular matrix components and modulates the activity of heparin-binding growth factors. The expression of syndecan-1 is modified during development, carcinogenesis, and tissue regeneration. During cutaneous wound healing, syndecan-1 expression is transiently induced in newly-formed capillaries of granulation tissue as well as in proliferating keratinocytes. To study the mechanisms underlying this regulation we investigated the effects of several growth factors/cytokines on syndecan-1 expression in two human cell lines: EA.hy 926 endothelial cells and HaCaT keratinocytes. None of these factors significantly altered syndecan-1 mRNA expression in cultured keratinocytes, but when given to endothelial cells, tumor necrosis factor-alpha (TNF-alpha) specifically and dose-dependently suppressed syndecan-1 expression at both mRNA and protein levels. TNF-alpha reduced the amount of syndecan-1 protein in EA.hy 926 cells in both the presence and absence of serum and, at the same time, induced the expression of intercellular adhesion molecule-1 (ICAM-1). The suppressive effect of TNF-alpha on endothelial syndecan 1 expression was reproducible in in vivo experiments in which TNF-alpha-coated beads were administered directly to healing skin wounds of mice. Data supporting these findings were further obtained by injecting TNF-alpha into an experimental rat granulation tissue model. In this tissue TNF-alpha suppressed syndecan-1 mRNA expression by approximately 80%. These results indicate that TNF-alpha is capable of down-regulating syndecan-1 expression in endothelial cells both in vitro and in vivo and suggest that similar mechanisms may be responsible for the changes in syndecan-1 expression observed during various regenerative, developmental, and malignant processes. PMID- 8702533 TI - Specific interaction between human kinetochore protein CENP-C and a nucleolar transcriptional regulator. AB - CENP-C is a human kinetochore protein that was originally identified as a chromosomal autoantigen in patients with scleroderma spectrum disease. To begin to establish a comprehensive protein map of the human centromere, affinity chromatography was used to identify nuclear proteins that specifically interact with CENP-C. Whereas a number of polypeptides appeared to interact with the full length CENP-C protein, only a pair of similarly sized proteins of approximately 100 kDa specifically interacted with the isolated carboxyl-terminal third of the CENP-C protein. Neither protein of the doublet bound to control affinity columns. Affinity purification and microsequence analysis of the proteins in the doublet identified them as the two highly related nucleolar transcription factors, UBF1 and UBF2 (also known as the nucleolar autoantigen NOR-90). Immunoblot analysis confirmed that both proteins also interacted with the full-length CENP-C polypeptide with similar affinities. Double indirect immunofluorescence using monospecific antibodies demonstrated that a subset of CENP-C and UBF/NOR-90 is colocalized at nucleoli of interphase HeLa cells, suggesting that the in vitro interaction detected by affinity chromatography may reflect an interaction that occurs in vivo. We discuss the implications of these findings in terms of the properties of interphase centromeres and the role of the nucleolus in scleroderma autoimmunity. PMID- 8702534 TI - Conserved region 3 of Escherichia coli final sigma70 is implicated in the process of abortive transcription. AB - Multiple-round in vitro transcription assays were performed using purified Escherichia coli RNA polymerase reconstituted with either wild-type or mutant final sigma70 proteins. These mutants, final sigma70(P504L) and final sigma70(S506F), bear single amino acid changes in conserved protein region 3. Behavior of the mutant enzymes on three test templates, bearing either the T7 A1, T5 N25, or T5 N25antiDSR promoter, were characterized. Transcription of all three promoter templates produced a pattern of specific abortive RNA species, which was qualitatively different for the mutants compared to the wild-type final sigma70 enzyme. Short abortive RNAs were produced at similar levels for mutant and wild type enzymes. The production of longer abortive species was either reduced or increased by the mutant enzymes in a systematic manner that appears promoter specific, and could be RNA length- or promoter distance-dependent. The process of abortive RNA transcription is thought to be tightly associated with that of promoter clearance. However, promoter clearance from these templates appears only slightly affected by the mutant enzymes. These mutants implicated region 3 of final sigma70 in the process of abortive transcription and suggest that the sequence of enzymatic events leading to the production of abortive or full-length RNA may be separable. PMID- 8702535 TI - The X-ray crystal structures of Yersinia tyrosine phosphatase with bound tungstate and nitrate. Mechanistic implications. AB - X-ray crystal structures of the Yersinia tyrosine phosphatase (PTPase) in complex with tungstate and nitrate have been solved to 2. 4-A resolution. Tetrahedral tungstate, WO42-, is a competitive inhibitor of the enzyme and is isosteric with the substrate and product of the catalyzed reaction. Planar nitrate, NO3-, is isosteric with the PO3 moiety of a phosphotransfer transition state. The crystal structures of the Yersinia PTPase with and without ligands, together with biochemical data, permit modeling of key steps along the reaction pathway. These energy-minimized models are consistent with a general acid-catalyzed, in-line displacement of the phosphate moiety to Cys403 on the enzyme, followed by attack by a nucleophilic water molecule to release orthophosphate. This nucleophilic water molecule is identified in the crystal structure of the nitrate complex. The active site structure of the PTPase is compared to alkaline phosphatase, which employs a similar phosphomonoester hydrolysis mechanism. Both enzymes must stabilize charges at the nucleophile, the PO3 moiety of the transition state, and the leaving group. Both an associative (bond formation preceding bond cleavage) and a dissociative (bond cleavage preceding bond formation) mechanism were modeled, but a dissociative-like mechanism is favored for steric and chemical reasons. Since nearly all of the 47 invariant or highly conserved residues of the PTPase domain are clustered at the active site, we suggest that the mechanism postulated for the Yersinia enzyme is applicable to all the PTPases. PMID- 8702536 TI - Involvement of Trp-284, Val-296, and Val-297 of the human delta-opioid receptor in binding of delta-selective ligands. AB - Given the high homology in amino acid sequence between the delta-opioid receptor and the two other types (mu and kappa), distinct residues in this receptor may confer its selectivity to some ligands. In order to identify molecular determinants in the human delta receptor responsible for the selectivity of delta selective ligands, two different delta/mu chimeras were constructed. In the first one, the delta sequence from the top of transmembrane 5 to the C terminus was replaced by the equivalent mu sequence, and in the second one, 13 consecutive residues in the third extracellular loop region of the delta receptor were replaced by the mu counterpart. These two chimeras retained the ability to bind the nonselective bremazocine but completely lost the ability to bind different delta-selective ligands. These results suggested that the region of the third extracellular loop of the delta receptor is crucial for the type selectivity. Furthermore, an alanine scan was performed by site-directed mutagenesis of 20 amino acids located in or proximal to the third extracellular loop. Among all the point mutations, only mutations of Trp-284, Val-296, or Val-297 significantly decreased the binding of delta-selective ligands tested. Moreover, combined mutation of Trp-284, Val-296, and Val-297 considerably decreased the affinities of the receptor for delta-selective ligands compared with the single point mutations. These findings suggest that Trp-284, Val-296, and Val-297 are crucial residues involved in the delta receptor type selectivity. PMID- 8702537 TI - Molecular determinants of the myristoyl-electrostatic switch of MARCKS. AB - MARCKS is a protein kinase C (PKC) substrate which binds calcium/calmodulin and actin, and which has been implicated in cell motility, phagocytosis, membrane traffic, and mitogenesis. MARCKS cycles on and off the membrane via a myristoyl electrostatic switch (McLaughlin, S., and Aderem, A.(1995) Trends Biochem. Sci. 20, 272-276). Here we define the molecular determinants of the myristoyl electrostatic switch. Mutation of the N-terminal glycine results in a nonmyristoylated form of MARCKS which does not bind membranes and is poorly phosphorylated. This indicates that myristic acid targets MARCKS to the membrane, where it is efficiently phosphorylated by PKC. A chimeric protein in which the N terminus of MARCKS is replaced by a sequence, which is doubly palmitoylated, is phosphorylated by PKC but not released from the membrane. Thus two palmitic acid moieties confer sufficient membrane binding energy to render the second, electrostatic membrane binding site superfluous. Mutation of the PKC phosphorylation sites results in a mutant which does not translocate from the membrane to the cytosol. A mutant in which the intervening sequence between the myristoyl moiety and the basic effector domain is deleted, is not displaced from the membrane by PKC dependent phosphorylation, fulfilling a theoretical prediction of the model. In addition to the nonspecific membrane binding interactions conferred by the myristoyl-electrostatic switch, indirect immunofluorescence microscopy demonstrates that specific protein-protein interactions also specify the intracellular localization of MARCKS. PMID- 8702538 TI - A single N-linked glycosylation site is implicated in the regulation of ligand recognition by the I-type lectins CD22 and CD33. AB - CD22 is an immunoglobulin superfamily B lymphocyte-specific adhesion receptor and a member of the recently identified I-type class of lectins. Recent work has shown that CD22 specifically recognizes sialic acid linked alpha2,6 to terminal N linked oligosaccharides on selected cell surface glycoproteins. CD22-ligand interaction is regulated by the activity of a beta-galactoside alpha2, 6 sialyltransferase that can inactivate CD22-mediated binding by sialylating the CD22 receptor itself. These observations suggest that N-linked glycosylation sites on the CD22 molecule may play a role in the regulation of CD22-mediated adhesion. In this work we have performed site-specific mutagenesis of potential N linked glycosylation sites on CD22 in an effort to determine whether they might be involved in ligand recognition. We show that mutation of a single potential N linked glycosylation site in the first immunoglobulin domain of CD22 completely abrogates ligand recognition. Interestingly, this site is characterized by the sequence NCT, where the cysteine is thought to be involved in an intrachain disulfide bond. Site-directed mutagenesis of similar NC(T/S) motifs in the first or second Ig domains of the I-type lectins myelin-associated glycoprotein, and sialoadhesin did not disrupt their ability to mediate sialic acid binding. In contrast, mutation of a NCS motif in the first Ig domain of the I-type lectin CD33 unmasked its sialic acid binding activity. These observations suggest that a single N-linked glycosylation site located at a similar position in the CD22 and CD33 glycoproteins is critical for regulating ligand recognition by both receptors. PMID- 8702539 TI - A possible predocking attachment site for N-ethylmaleimide-sensitive fusion protein. Insights from in vitro endosome fusion. AB - N-Ethylmaleimide-sensitive fusion protein (NSF) is an ubiquitous protein required for multiple vesicular transport events. We have investigated the role of the two nucleotide-binding regions of NSF in endosomal fusion by analyzing NSF mutants in a cell-free system. Our results indicate that mutations on the first ATP-binding domain, that render a protein defective in either ATP binding or ATP hydrolysis, results in almost complete inhibition of endosomal fusion. A mutation in the second ATP-binding site of NSF was only slightly inhibitory. The inhibitory effect was observed only when the mutant proteins were added at early times during the fusion reaction indicating that NSF may be required for an early step during the docking/fusion process. Binding studies using Western blotting reveal that the binding of NSF mutants to endosomal membranes is differentially affected by Ca2+. Our results indicate that NSF, depending on its nucleotide state, may interact with membranes via an alternate mechanism. Our findings suggest the existence of a predocking binding site either independent of the docking complex or a site that leads to the formation of the SNAP-SNARE complex (e.g. 20 S particle). PMID- 8702540 TI - Core histones are glutaminyl substrates for tissue transglutaminase. AB - Chicken erythrocyte core histones are glutaminyl substrates in the transglutaminase (TGase) reaction with monodansylcadaverine (DNC) as donor amine. The modification is very fast when compared with that of many native substrates of TGase. Out of the 18 glutamines of the four histones, nine (namely glutamine 95 of H2B; glutamines 5, 19, and 125 of H3; glutamines 27 and 93 of H4; and glutamines 24, 104, and 112 of H2A) are the amine acceptors in free histones. The use of Gln112 of H2A requires a temperature-dependent partial unfolding of the histone, showing that structural determinants are decisive for the glutamine specificity. The structures of H2A and H2B do not appreciably change upon modification with DNC as determined by circular dichroism, and core particles reconstituted from these DNC-modified histones are indistinguishable from native nucleosome cores. When the reaction is carried out with native nucleosomes, only glutamines 5 and 19 of H3, which are located in the N-terminal tail, and glutamine 22 of H2B, which is not labeled in free histone, are modified. Methylamine and putrescine also are incorporated into nucleosomes by the TGase reaction. Our results reveal several possibilities for the application of the TGase reaction in the chromatin field, and taking into account that histones are easily cross-linked or modified by polyamines in vitro, the possibility that they may be TGase substrates in vivo is discussed. PMID- 8702541 TI - The major calpain isozymes are long-lived proteins. Design of an antisense strategy for calpain depletion in cultured cells. AB - Calpains are intracellular Ca2+-dependent proteases that are thought to participate in Ca2+-associated signal transduction pathways. It has been proposed that calpains are activated by an autoproteolytic mechanism. If this is true one would expect a relatively short half-life for calpain protein in cells. To test this hypothesis, WI-38 human diploid fibroblasts were pulse-labeled with [35S]methionine, and calpain was immunoprecipitated at various times after chasing with nonradioactive methionine to determine residual radioactivity. The results demonstrated that the two major calpain isozymes, m-calpain and micro calpain, had metabolic half-lives of approximately 5 days. Calpains were long lived proteins in several human cell lines, A-431, HeLa, VA-13, C-33A, and TE2 cells. In addition, calpastatin, the calpain-specific inhibitor protein, also had a long metabolic half-life. These observations suggest that the model for calpain activation by autoproteolysis requires re-investigation. Based on a knowledge of calpain metabolic stability, a protocol was devised for chronic exposure of WI-38 cells and HeLa cells to a calpain small subunit antisense oligodeoxyribonucleotide. Depletion of calpain small subunit after 5 or more days of treatment led to inhibition of cell proliferation that could be reversed by removal of antisense oligodeoxyribonucleotide from the culture medium. Together with previous studies, these results indicate a requirement for calpains in mammalian cell proliferation. PMID- 8702542 TI - Conversion of product specificity of archaebacterial geranylgeranyl-diphosphate synthase. Identification of essential amino acid residues for chain length determination of prenyltransferase reaction. AB - Prenyltransferases catalyze the consecutive condensation of isopentenyl diphosphate with allylic diphosphates to produce prenyl diphosphates whose chain lengths are absolutely determined by each enzyme. To investigate the mechanism of the consecutive reaction and the determination of the ultimate chain length, a random mutational approach was planned. A geranylgeranyl-diphosphate synthase gene from Sulfolobus acidocaldarius was randomly mutagenized by NaNO2 treatment to construct a library of mutated geranylgeranyl-diphosphate synthase genes on a yeast expression vector. The library was screened for suppression of a pet phenotype of yeast C296-LH3, which is deficient in hexaprenyl-diphosphate synthase. Five mutants that could grow on a YEPG plate, which contained only glycerol as an energy source instead of glucose, were selected from approximately 1,400 mutants. All selected mutated enzymes catalyzed the formation of polyprenyl diphosphates with prenyl chains longer than geranylgeranyl diphosphate. Especially mutants 1, 3, and 5 showed the strongest elongation activity to produce large amounts of geranylfarnesyl diphosphate with a concomitant amount of hexaprenyl diphosphate. Sequence analysis revealed that each mutant contained a few amino acid substitutions and that the mutation of Phe-77, which is located on the fifth amino acid upstream from the first aspartate-rich consensus motif, is the most effective for elongating the ultimate product. Amino acid alignment of known prenyltransferases around this position and our previous observations on farnesyl-diphosphate synthase (Ohnuma, S.-i., Nakazawa, T., Hemmi, H., Hallberg, A.-M., Koyama, T., Ogura, K., and Nishino, T.(1996) J. Biol. Chem. 271, 10087 10095) clearly indicate that the amino acid at the position of all prenyltransferases must regulate the chain elongation. PMID- 8702543 TI - Structure-function relationships of the mouse Gap1m. Determination of the inositol 1,3,4,5-tetrakisphosphate-binding domain. AB - Gap1(IP4BP), one of a member of Ras GTPase-activating proteins, has been identified as a specific inositol 1,3,4,5-tetrakisphosphate (IP4)-binding protein (Cullen, P. J., Hsuan, J. J., Truong, O., Letcher, A. J., Jackson, T. R., Dawson, A. P., and Irvine, R. F. (1995) Nature 386, 527-530). In this paper we describe Gap1(m), which is closely related to Gap1(IP4BP), to also be an IP4-binding protein and show that the pleckstrin homology domain (PH) is the central IP4 binding domain by expressing fragments of the mouse Gap1(m) in Escherichia coli as fusion proteins and examining their activities. However, in addition to the PH domain, an adjacent GAP-related domain and carboxyl terminus are required for high affinity specific IP4 binding. The PH domain is highly conserved in the Gap1 family and also has striking homology to the amino-terminal region of Bruton's tyrosine kinase. Substitution of Cys for Arg at position 628 in the PH domain corresponding to the mutation of Bruton's tyrosine kinase observed in X-linked immunodeficiency mice results in a dramatic reduction of IP4 binding activity as well as phospholipid binding capacity of Gap1(m). This mutant also showed the GAP activity against Ha-Ras to be similar to that of the wild type Gap1(m). Our results suggest that the PH domain of Gap1(m) functions as a modulatory domain of GAP activity by binding IP4 and phospholipids. PMID- 8702544 TI - Subunit structure and organization of the genes of the A1A0 ATPase from the Archaeon Methanosarcina mazei Go1. AB - The proton-translocating A1A0 ATP synthase/hydrolase of Methanosarcina mazei Go1 was purified and shown to consist of six subunits of molecular masses of 65, 49, 40, 36, 25, and 7 kDa. Electron microscopy revealed that this enzyme is organized in two domains, the hydrophilic A1 and the hydrophobic A0 domain, which are connected by a stalk. Genes coding for seven hydrophilic subunits were cloned and sequenced. From these data it is evident that the 65-, 49-, 40- and 25-kDa subunits are encoded by ahaA, ahaB, ahaC, and ahaD, respectively; they are part of the A1 domain or the stalk. In addition there are three more genes, ahaE, ahaF, and ahaG, encoding hydrophilic subunits, which were apparently lost during the purification of the protein. The A0 domain consists of at least the 7-kDa proteolipid and the 36-kDa subunit for which the genes have not yet been found. In summary, it is proposed that the A1A0 ATPase of Methanosarcina mazei Go1 contains at least nine subunits, of which seven are located in A1 and/or the stalk and two in A0. PMID- 8702545 TI - Two different cytoplasmic tails direct isoforms of the membrane cofactor protein (CD46) to the basolateral surface of Madin-Darby canine kidney cells. AB - Membrane cofactor protein (MCP; CD46), a widely distributed regulatory protein of the complement system, was analyzed for expression in polarized epithelial cells. Both a human and a simian (Vero C1008) cell line were found to contain endogenous MCP mainly on the basolateral surface. Transfected Madin-Darby canine kidney cells stably expressing human MCP delivered this protein also predominantly to the basolateral surface. A deletion mutant lacking the cytoplasmic tail was transported in a nonpolarized fashion, indicating that the targeting signal for the basolateral transport is located in the cytoplasmic domain. A characteristic feature of MCP is the presence of various isoforms that contain either of two different cytoplasmic tails as a consequence of alternative splicing. Two isoforms differing only in the cytoplasmic tail (tail 1 or 2) were analyzed for polarized expression in Madin-Darby canine kidney cells. Surface biotinylation, as well as confocal immunofluorescence microscopy, indicated that both proteins were transported to the basolateral surface. Because no sequence similarity has been observed, the two tails contain different basolateral targeting signals. A deletion mutant lacking the only tyrosine residue in tail 1 retained the polarized expression indicating that, in contrast to most basolateral sorting signals, the transport signal of the tail 1 isoform is not dependent on tyrosine. The maintenance of a targeting motif in two distinct cytoplasmic tails suggests that the basolateral expression of MCP in polarized epithelial cells is of physiological importance. PMID- 8702547 TI - Intramolecular disulfide bonds between conserved cysteines in wheat gliadins control their deposition into protein bodies. AB - Following synthesis, wheat gliadin storage proteins are deposited into protein bodies inside the endomembrane system in a way that enables not only their efficient accumulation and dehydration during seed maturation, but also their rapid rehydration and degradation during germination. In the present report, we studied the mechanism of gliadin deposition and whether it was controlled by the conformation of these proteins. Although gliadins are generally known to be insoluble in aqueous solutions, sucrose gradient analysis showed that a considerable amount of these proteins appeared as relatively soluble monomers in developing grains. In vitro reduction of the intramolecular disulfide bonds that are present in natural monomeric gliadins caused their precipitation into insoluble aggregates. In addition, pulse-chase experiments in the absence or presence of reducing agents showed that formation of intramolecular disulfide bonds also played a major role in folding and deposition of the gliadins in vivo. Our results imply that following sequestration into the endoplasmic reticulum, the gliadins fold into relatively soluble monomers, which are incompetent for rapid aggregation and gradually assemble into protein bodies. This pattern of deposition apparently depends on the conformation of the gliadins, which is stabilized by intramolecular disulfide bonds formed between the conserved cysteines. The contribution of this study to the understanding of the evolution and function of gliadins is discussed. PMID- 8702546 TI - Identification and cloning of centaurin-alpha. A novel phosphatidylinositol 3,4,5 trisphosphate-binding protein from rat brain. AB - Using an affinity resin and photoaffinity label based on phospholipid analogs of inositol 1,3,4,5-tetrakisphosphate (InsP4), we have isolated, characterized, and cloned a 46-kDa protein from rat brain, which we have named centaurin-alpha. Binding specificity was determined using displacement of 1-O-[3H](3-[4 benzoyldihydrocinnamidyl]propyl)-InsP4 photoaffinity labeling. Centaurin-alpha displayed highest affinity for phosphatidylinositol 3,4,5-trisphosphate (PtdInsP3) (IC50 = 120 nM), whereas InsP4, PtdInsP2, and InsP3 bound with 5-, 12 , and >50-fold lower affinity, respectively. Screening a rat brain cDNA library with a polymerase chain reaction product, generated using partial amino acid sequence from tryptic peptides, yielded a full-length clone. The 2,450-base pair cDNA contained an open reading frame (ORF) encoding a novel protein of 419 amino acids. Northern analysis revealed a 2.5-kilobase transcript that is highly expressed in brain. The deduced sequence contains a novel putative zinc finger motif, 10 ankyrin-like repeats, and shows homology to recently identified yeast and mammalian Arf GTPase-activating proteins. Given the specificity of binding and enrichment in brain, centaurin-alpha is a candidate PtdInsP3 receptor that may link the activation of phosphoinositide 3-kinase to downstream responses in the brain. PMID- 8702548 TI - Anti-IgM-mediated regulation of c-myc and its possible relationship to apoptosis. AB - Anti-IgM treatment of Burkitt's lymphoma cells is followed by either growth arrest or induction of apoptosis. In this study we have explored the role of c myc in these events. Our results in Ramos cells indicate the following. (a) The decline in c-myc mRNA occurs at about 4 h; inhibition of about 80% being observed. (b) The stability of c-myc message is involved since the half-life of c myc mRNA is decreased from about 30 min in untreated cells to about 15 min following treatment with anti-IgM. In the presence of cycloheximide, a protein synthesis inhibitor, the half-life is increased to about 50 min and was unaltered by treatment with anti-IgM. (c) By contrast, nuclear run-on experiments indicated no change in transcription rates for c-myc message due to treatment with anti IgM. (d) A decrease in c-myc causes apoptosis since specific repression of c-myc with antisense oligonucleotides decreases the levels of c-Myc, inhibits growth rate, decreases viability, and induces apoptosis. (e) Anti-CD40 inhibition of apoptosis occurs without alteration in anti-IgM-induced down-regulation of c-myc mRNA, suggesting that it acts distally to c-myc down-regulation. Other cell lines were also investigated. In Epstein-Barr virus (EBV)-positive cell lines (Daudi, Raji, and Namalwa), anti-IgM treatment for 24 h results in growth inhibition without induction of apoptosis. In EBV-negative cell lines (ST486 and CA46, as well as Ramos), a more heterogeneous pattern of responses to anti-IgM are observed. Ramos and ST486 cells both show growth inhibition and apoptosis upon anti-IgM treatment; CA46 cells shown only growth inhibition but not apoptosis. Anti-IgM causes a decline in c-myc mRNA levels in all of these lines, as well as in c-Myc protein level in the two lines investigated, Daudi and Ramos, regardless of apoptosis. Addition of antisense c-myc oligonucleotides to the cells reduced growth in both Daudi and Ramos cells lines, however it resulted in substantial apoptosis only in Ramos cells. These results suggest that anti-IgM destabilizes c myc mRNA by a process that involves mRNA turnover, rather than transcription rates. However anti-IgM exerts differential effects in EBV-positive and EBV negative cell lines. EBV-positive cells are uniformly resistant to apoptosis, while EBV-negative cell lines show a tendency to apoptosis but with exceptions. Growth inhibition can be uncoupled from apoptosis in EBV-positive cell lines, but not in those EBV-negative cell lines prone to apoptosis. Furthermore, down regulation of c-myc message correlates with growth inhibition in these cells, but is an insufficient link to apoptosis. By contrast inhibition of apoptosis by anti CD40 occurs even though c-myc mRNA is decreased. PMID- 8702549 TI - Antisense RNA, fur, iron, and the regulation of iron transport genes in Vibrio anguillarum. AB - The negative regulation of the expression of iron transport genes fatA and fatB in Vibrio anguillarum is mediated by a chromosome-encoded Fur protein and a plasmid pJM1-derived antisense RNA (RNAalpha), which is preferentially expressed under iron-rich conditions. In this work, we characterized the RNAalpha promoter region, and by using promoter fusion and rifampicin experiments we were able to demonstrate that iron regulates RNAalpha synthesis posttranscriptionally by stabilizing RNAalpha half-life rather than enhancing transcription initiation. The Fur protein is also essential for RNAalpha synthesis at the transcription initiation level, independently of the iron status of the cell. From experiments assessing the relative contribution of Fur and RNAalpha, we were able to show that RNAalpha may indeed play an important role on the negative regulation of the expression of the iron transport genes under physiological conditions. PMID- 8702550 TI - Identification of a family of streptococcal surface proteins with extremely repetitive structure. AB - The group B Streptococcus (GBS) causes the majority of life-threatening bacterial infections in newborn children. Most GBS strains isolated from such infections express a surface protein, designated Rib, that confers protective immunity and therefore is of interest for analysis of pathogenetic mechanisms. Sequence analysis demonstrated that Rib has an exceptionally long signal peptide (55 amino acid residues) and 12 repeats (79 amino acid residues each) that account for >80% of the sequence of the mature protein. The repeats are identical even at the DNA level, indicating that an efficient mechanism operates to maintain a highly repetitive structure in Rib. The structure of Rib is similar to that of alpha, a previously characterized surface protein that is common among GBS strains lacking Rib. However, highly purified preparations of Rib and alpha did not cross-react immunologically, although the two proteins show extensive amino acid residue identity (47% in the repeat region). When analyzed in Western blots, Rib and alpha give rise to a regularly spaced ladder pattern, apparently due to hydrolysis of acid-labile Asp-Pro bonds in the repeats. We conclude that Rib and alpha are members of a novel family of streptococcal surface proteins with unusual repetitive structure. PMID- 8702551 TI - Manganese superoxide dismutase modulates interleukin-1alpha levels in HT-1080 fibrosarcoma cells. AB - Reactive oxygen species of mitochondrial origin have been implicated in regulating the expression of several tumor necrosis factor (TNF)-induced genes. Manganese superoxide dismutase (Mn-SOD) is one of many genes, but only antioxidant enzyme, induced in response to tumor necrosis factor. Mn-SOD is a nuclear-encoded mitochondrial matrix protein and serves a protective function by detoxifying superoxide. To address the role of superoxide in regulating gene expression in response to TNF, we have constitutively overexpressed Mn-SOD in a human fibrosarcoma cell line and asked what effect this has on the expression of a number of TNF-responsive genes using reverse transcription-polymerase chain reaction. Of the TNF-induced transcripts analyzed, only interleukin-1alpha (IL 1alpha) was modulated in response to Mn-SOD overexpression. In all cases of Mn SOD overexpression, IL-1alpha protein and mRNA levels were lowered constitutively and in response to TNF when compared to the parental and mock-transfected cell lines. The induction of IL-1alpha by TNF can also be decreased by growth in 3% oxygen as compared to growth in 21% O2; in addition, growth in low oxygen lowers the basal level of IL-1alpha protein. The effect of Mn-SOD overexpression on IL 1alpha expression can be overcome by treatment with the protein kinase C activator, phorbol 12-myristate 13-acetate. Mn-SOD overexpression and low oxygen alter IL-1alpha mRNA levels by decreasing the stability of the IL-1alpha mRNA. These findings indicate that both Mn-SOD and O2 may regulate the levels of a cellular oxidant involved in both basal and TNF-induced IL-1alpha expression, presumably superoxide. PMID- 8702552 TI - Voltage-gated K+ channels contain multiple intersubunit association sites. AB - A domain in the cytoplasmic NH2 terminus of voltage-gated K+ channels supervises the proper assembly of specific tetrameric channels (Li, M., Jan, J. M., and Jan, L. Y.(1992) Science 257, 1225-1230; Shen, N. V., Chen X., Boyer, M. M., and Pfaffinger, P. (1993) Neuron 11, 67-76). It is referred to as a first tetramerization domain, or T1 (Shen, N. V., Chen X., Boyer, M. M., and Pfaffinger, P.(1993) Neuron 11, 67-76). However, a deletion mutant of Kv1.3 that lacks the first 141 amino acids, Kv1.3 (T1(-)) forms functional channels, suggesting that additional association sites in the central core of Kv1.3 mediate oligomerization. To characterize these sites, we have tested the abilities of cRNA Kv1.3 (T1(-)) fragments co-injected with Kv1.3 (T1(-)) to suppress current in Xenopus oocytes. The fragments include portions of the six putative transmembrane segments, S1 through S6, specifically: S1, S1-S2, S1-S2-S3, S2-S3, S2-S3-S4, S3-S4, S3-S4-S5, S2 through COOH, S3 through COOH, S4 through COOH, and S5-S6-COOH. Electrophysiologic experiments show that the fragments S1-S2-S3, S3 S4-S5, S2 through COOH, and S3 through COOH strongly suppress Kv1.3 (T1(-)) current, while others do not. Suppression of expressed current is due to specific effects of the translated peptide Kv1.3 fragments, as validated by in vivo immunoprecipitation studies of a strong suppressor and a nonsuppressor. Pulse chase experiments indicate that translation of truncated peptide fragments neither prevents translation of Kv1.3 (T1(-)) nor increases its rate of degradation. Co-immunoprecipitation experiments suggest that suppression involves direct association of a peptide fragment with Kv1.3 (T1(-)). Fragments that strongly suppress Kv1.3 (T1(-)) also suppress an analogous NH2-terminal deletion mutant of Kv2.1 (Kv2.1 (DeltaN139)), an isoform belonging to a different subfamily. Our results indicate that sites in the central core of Kv1.3 facilitate intersubunit association and that there are suppression sites in the central core, which are promiscuous across voltage-gated K+ channel subfamilies. PMID- 8702554 TI - RGFGIGS is an amino acid sequence required for acetyl coenzyme A binding and activity of human spermidine/spermine N1acetyltransferase. AB - Polyamine catabolism is rate limited by spermidine/spermine N1-acetyltransferase (SSAT). Although the amino acid sequence of SSAT is known, the substrate binding and catalytic sites are not. The goal of this study was to define the region responsible for acetyl coenzyme A binding. Human SSAT contains a region of 20 amino acids homologous to several microbial antibiotic N-acetyltransferases. The highest homology is represented in the Campylobacter coli streptothricin acetyltransferase sat4 gene, where 16 identical or highly conserved amino acids exist in a 20-residue stretch. The most conserved residues within this region are RGFGIGS beginning at Arg-101 in the human SSAT. Site-directed mutations to Arg 101, Gly-104, and Gly-106 resulted in proteins with no measurable activity. The G102D mutation produced a partially active protein with a decreased affinity for acetyl coenzyme A and with a Km >10-fold that of the wild-type protein. Analysis using the PredictProtein program suggests a common structure among the microbial and eukaryotic N-acetyltransferases in the region corresponding to the RGFGIGS of human SSAT consisting of an alpha-helix usually preceded by a glycine loop. Our data are consistent with the hypothesis that Arg-101 and the proximal glycine loop are necessary for the activity of human SSAT. PMID- 8702555 TI - Sp1 activates and inhibits transcription from separate elements in the proximal promoter of the human adenine nucleotide translocase 2 (ANT2) gene. AB - Expression of the adenine nucleotide translocator 2 (ANT2) gene is growth regulated. We report a feature of the ANT2 promoter that involves a novel regulatory function for the Sp1 transfactor. We show that expression from the ANT2 proximal promoter is modulated through three Sp1 elements, two of which activate and one of which partially inhibits transcription. The inhibitor site, box C, is juxtaposed to transcription start (nucleotides -7 to -2). Sp1 bound to box C decreases transcription initiation. This was demonstrated by introducing mutations in box C which (a) increased chloramphenicol acetyltransferase expression in the transient transfection assay and (b) inhibited binding of both purified Sp1 and Sp1 in crude nuclear extracts. The activating elements (A and B boxes) are located at adjacent sites in the distal region of the proximal promoter. Mutation of either box inhibits transfection by 90%, indicating that they act in a synergistic manner. Supershift experiments with crude nuclear extracts showed that only Sp1 was bound to the three GC boxes. The finding that Sp1 acts as an activator/inhibitor within the same promoter region was verified in NIH3T3, HeLa, JEG3, and COS-1, indicating that this dual effect of Sp1 is widely preserved. These data suggest a unique role for Sp1 and raise the possibility that growth activation of the ANT2 gene is regulated by the interaction of Sp1 on the A, B, and C boxes. PMID- 8702556 TI - Identification and cDNA cloning of 35-kDa phosphatidic acid phosphatase (type 2) bound to plasma membranes. Polymerase chain reaction amplification of mouse H2O2 inducible hic53 clone yielded the cDNA encoding phosphatidic acid phosphatase. AB - We previously described the purification of an 83-kDa phosphatidic acid phosphatase (PAP) from the porcine thymus membranes (Kanoh, H., Imai, S.-i., Yamada, K. and Sakane, F.(1992) J. Biol. Chem. 267, 25309-25314). However, we found that a minor 35-kDa protein could account for the PAP activity when the purified enzyme preparation was further analyzed. We thus determined the N terminal sequence of the 35-kDa candidate protein and prepared antipeptide antibody against the determined sequence, MFDKTRLPYVALDVL. The antibody almost completely precipitated the purified enzyme activity. Furthermore, the antibody precipitated from the radioiodinated enzyme preparation a single 35-kDa protein, which was converted to a 29-kDa form when treated with N-glycanase. We also found that the immunoprecipitable PAP activity was exclusively associated with the plasma membranes of porcine thymocytes. These results indicated that the 35-kDa glycosylated protein represents the plasma membrane-bound (type 2) PAP. We surprisingly noted that the N-terminal sequence of the porcine PAP was almost completely conserved in the internal sequence encoded by a mouse partial cDNA clone, hic53, reported as a H2O2-inducible gene (Egawa, K., Yoshiwara, M., Shibanuma, M., and Nose, K.(1995) FEBS Lett. 372, 74-77). We thus amplified from the mouse kidney RNA the hic53 clone by polymerase chain reaction, and obtained a cDNA encoding a novel protein of 283 amino acid residues with a calculated Mr of 31,894. Methionine reported as an internal residue was found to serve as an initiator, and the C-terminal 64 residues were lacking in hic53. The protein contains several putative membrane-spanning domains and two N-glycosylation sites. When transfected into 293 cells, the cDNA gave more than 10-fold increase of the membrane-bound PAP activity, which could be precipitated by the antipeptide antibody. In [35S]methionine-labeled cells, the translational product was confirmed to be a 35-kDa protein, which became 30 kDa in cells treated with tunicamycin, an inhibitor of N-glycosylation. We thus succeeded first in identifying the porcine type 2 PAP and subsequently in determining the primary structure of a mouse homolog of the PAP. PMID- 8702557 TI - Effects of Xenopus laevis mitochondrial single-stranded DNA-binding protein on primer-template binding and 3'-->5' exonuclease activity of DNA polymerase gamma. AB - Mitochondrial DNA (mtDNA) is replicated by DNA polymerase gamma by a strand displacement mechanism involving mitochondrial single-stranded DNA-binding protein (mtSSB). mtSSB stimulates the overall rate of DNA synthesis on singly primed M13 DNA mainly by stimulating the processivity of DNA synthesis rather than by stimulating primer recognition. We used electrophoretic mobility shift methods to study the effects of mtSSB on primer-template recognition by DNA pol gamma. Preliminary experiments showed that single mtSSB tetramers bind tightly to oligo(dT) single strands containing 32 to 48 residues. An oligonucleotide primer template was designed with an 18-mer primer annealed to the 3'-portion of a 71 mer template containing 40 dT residues at its 5'-end as a binding site for mtSSB. DNA pol gamma bound to this primer-template either in the absence or presence of mtSSB in complexes that remained intact and enzymatically active following native gel electrophoresis. Association of mtSSB with the 5'-dT40-tail in the 18:71-mer primer-template reduced the binding of DNA polymerase gamma and the efficiency of primer extension. Binding of mtSSB to single-stranded DNA was also observed to block the action of the 3'-->5' exonuclease of DNA polymerase gamma. The size of fragments protected from 3'-->5' exonuclease trimming increases with increasing ionic strength in a manner consistent with the known salt dependence of the binding site size of Escherichia coli SSB. PMID- 8702558 TI - Fidelity and error specificity of the alpha catalytic subunit of Escherichia coli DNA polymerase III. AB - Escherichia coli DNA polymerase III holoenzyme is the replicative enzyme primarily responsible for the duplication of the E. coli chromosome. This process occurs with high accuracy, less than 10(-9) to 10(-10) errors being committed per base pair per round of replication. As a first step in understanding the mechanisms responsible for the high fidelity of this process, we have purified the polymerase III alpha catalytic subunit, free of exonuclease activity, and analyzed its fidelity in vitro. We employed a newly developed gap-filling assay using the N-terminal 250 bases of the lacI gene as a forward mutational target. When synthesizing across this target, alpha subunit produced mutations at a frequency of 0.6%. DNA sequencing revealed that the mutants created in vitro consisted mostly of frameshift mutations, although some base substitutions were also observed. The frameshifts, occurring at more than 120-fold above the background, consisted largely of -1 deletions. Among them, about 80% were the deletion of a purine template base with a pyrimidine 5'-neighbor. These results suggest that the alpha subunit (i) has a relatively low ability to extend from misincorporated bases, accounting for the low level of observed base substitutions, and (ii) has a relatively high capability of extension after misalignment of a misincorporated base on the next (complementary) template base, accounting for the high level of frameshift mutations. This model is supported by an experiment in which alpha subunit was required to initiate DNA synthesis from a terminal mispair in a sequence context that allowed slippage on the next template base. Among the products of this reaction, frameshifts outnumbered base pair substitutions by greater than 70-fold. A comparison to in vivo mutational spectra suggests that the pol III accessory factors may play a major role in modulating the fidelity of DNA synthesis. PMID- 8702559 TI - Fibronectin mRNA splice variant in articular cartilage lacks bases encoding the V, III-15, and I-10 protein segments. AB - Fibronectin is an extracellular matrix glycoprotein encoded by a single gene. Alternative RNA splicing has been reported at three sites, ED (extra type III domain)-A, ED-B, and the variable or V region. Articular cartilage fibronectin monomers are rarely (ED-A)+, but approximately 25% are (ED-B)+. RNA gel electrophoresis and Northern blot analysis identified two (ED-B)+ and two (ED-B)- fibronectin transcripts in cartilage, each pair differing by approximately 750 bases. This difference results from a previously unreported RNA splicing pattern that eliminates not only the V region but also nucleotides encoding protein segments III-15 and I-10. This new splice variant, which we designate (V+C)-, represents the majority of fibronectin transcripts in equine, canine, and rabbit articular cartilage but is absent in the liver. Reverse transcriptase-polymerase chain reaction analyses of 11 additional equine tissues failed to detect the (V+C)- splice variant, except for very low levels in lymph node, bone, aorta, and skin. Furthermore, chondrocytes grown in monolayer culture maintain high levels of fibronectin expression but stop expressing (V+C)- transcripts over time. The tissue-specific expression pattern of this novel fibronectin isoform suggests that it may have an important function in the matrix organization of cartilage. PMID- 8702560 TI - Decorin-induced growth suppression is associated with up-regulation of p21, an inhibitor of cyclin-dependent kinases. AB - The secreted proteoglycan decorin has been implicated in the negative control of cell proliferation primarily by virtue of its ability to block transforming growth factor-beta. Moreover, decorin expression is markedly up-regulated during quiescence but suppressed upon viral transformation, whereas de novo decorin expression in colon carcinoma cells abrogates the malignant phenotype by arresting the cells in the G1 phase of the cell cycle. Here we show that this decorin-induced growth arrest is associated with up-regulation of p21 mRNA and protein in a transforming growth factor-beta- and p53-independent pathway. The augmented p21 protein is present as a multimeric complex with various cyclins and cyclin-dependent kinases in the nuclei of decorin-expressing cells, thereby leading to suppression of cyclin-dependent kinase activity and block of cell division. Through the usage of decorin-specific antisense oligodeoxynucleotide treatment, we demonstrate that the expression of decorin is closely linked to that of p21 and that abrogation of decorin leads to suppression of p21 and restoration of cell division. Collectively, our results provide a plausible mechanism by which decorin may contribute to retard and suppress the growth of tumor cells in vivo. PMID- 8702561 TI - In vitro reconstitution of transcriptional antitermination by the SacT and SacY proteins of Bacillus subtilis. AB - Expression of the sacPA and sacB genes of Bacillus subtilis is positively modulated by transcriptional regulatory proteins encoded by the sacT and sacY genes, respectively. Previous genetic studies led to the suggestion that SacT and SacY function as nascent mRNA binding proteins preventing early termination of transcription at terminators located in the leader regions of the corresponding genes. Here we report the overproduction, purification to near homogeneity, and characterization of the two antiterminators, SacT and SacY. Using mRNA band migration retardation assays and a reconstituted transcriptional antitermination system, the mRNA binding functions and antitermination activities of purified SacT and SacY are demonstrated under in vitro conditions. The results establish for the first time that members of the BglG family of antiterminators function in antitermination in the absence of other proteins in vitro. Purified SacT is shown to be phosphorylated by phosphoenolpyruvate in a phosphotransferase-catalyzed reaction dependent on Enzyme I and HPr. Unexpectedly, the purified SacT is shown to be functional in mRNA binding and in transcriptional antitermination independently of its phosphorylation state. PMID- 8702562 TI - Isolation and characterization of Pas2p, a peroxisomal membrane protein essential for peroxisome biogenesis in the methylotrophic yeast Pichia pastoris. AB - The pas2 mutant of the methylotrophic yeast Pichia pastoris is characterized by a deficiency in peroxisome biogenesis. We have cloned the PpPAS2 gene by functional complementation and show that it encodes a protein of 455 amino acids with a molecular mass of 52 kDa. In a Pppas2 null mutant, import of both peroxisomal targeting signal 1 (PTS1)- and PTS2-containing proteins is impaired as shown by biochemical fractionation and fluorescence microscopy. No morphologically distinguishable peroxisomal structures could be detected by electron microscopy in Pppas2 null cells induced on methanol and oleate, suggesting that PpPas2p is involved in the early stages of peroxisome biogenesis. PpPas2p is a peroxisomal membrane protein (PMP) and is resistant to extraction by 1 M NaCl or alkaline sodium carbonate, suggesting that it is a peroxisomal integral membrane protein. Two hydrophobic domains can be distinguished which may be involved in anchoring PpPas2p to the peroxisomal membrane. PpPas2p is homologous to the Saccharomyces cerevisiae Pas3p. The first 40 amino acids of PpPas2p, devoid of the hydrophobic domains, are sufficient to target a soluble fluorescent reporter protein to the peroxisomal membrane, with which it associates tightly. A comparison with the membrane peroxisomal targeting signal of PMP47 of Candida boidinii revealed a stretch of positively charged amino acids common to both sequences. The role of peroxisomal membrane targeting signals and transmembrane domains in anchoring PMPs to the peroxisomal membrane is discussed. PMID- 8702563 TI - Activation of a nuclear DNA-binding protein recognized by a transcriptional element, bcn-1, from the laminin B2 chain gene promoter. AB - Treatment of mesangial cells with either phorbol 12-myristate 13-acetate (PMA) or interleukin-1beta induces an increase in laminin B2 chain mRNA levels. In other systems, activation of gene expression by these agents is transcriptionally mediated. To identify transcription factors that control expression of laminin B2 chain gene, we employed a strategy consisting of a computer-based analysis of murine and human gene promoter sequences and gel shift assays. Although overall the laminin B2 chain promoters from the two species have low sequence similarity, the mouse promoter contained sequences that were also contained in one motif, 5' CCCCGCCCACCTCGCGCGC-3', designated bcn-1, from the human promoter. Treatment of mesangial cells with either PMA or interleukin-1beta induced a transient increase in nuclear DNA binding activity, designated BCN-1, recognized the bcn-1 motif in a gel shift assay. A single nucleotide replacement in the bcn-1 motif abolished DNA binding, indicating that bcn-1.BCN-1 complex formation is highly specific. In transient transfections, the ability of PMA to induce the laminin B2 chain promoter was abolished by mutating the bcn-1 motif. These results suggest that the bcn-1 element and its cognate inducible BCN-1 protein regulate laminin B2 chain gene transcription. PMID- 8702564 TI - Selective cleavage of the heregulin receptor ErbB-4 by protein kinase C activation. AB - The 180-kDa transmembrane tyrosine kinase ErbB-4 is a receptor for the growth factor heregulin. 125I-Heregulin binding to NIH 3T3 cells overexpressing the ErbB 4 receptor is rapidly decreased by 12-O-tetradecanoylphorbol-13-acetate (TPA) pretreatment. Immunologic analysis demonstrates that TPA treatment of cells induces the proteolytic cleavage of ErbB-4, producing an 80-kDa cytoplasmic domain fragment, which contains a low level of phosphotyrosine, and a 120-kDa ectodomain fragment, which is released into the extracellular medium. Cleavage of ErbB-4 was also enhanced by other protein kinase C activators, i.e. platelet derived growth factor, ionomycin, and synthetic diacylglycerol, while protein kinase C inhibition or down-regulation suppressed the TPA stimulation of ErbB-4 degradation. TPA did not induce the degradation of related receptors (ErbB-1, ErbB-2, and ErbB-3) in the EGF receptor family. The phorbol ester-induced cleavage of ErbB-4 occurs within or close to the ectodomain, as the 80-kDa cytoplasmic domain fragment is recognized by antibody to the ErbB-4 carboxyl terminus and is membrane-associated. Coprecipitation experiments show that, while the 80-kDa ErbB-4 fragment is associated with the SH2-containing molecules PLC gamma1 and Shc, TPA did not induce the phosphorylation of these substrates in intact cells. In addition, kinase assays in vitro indicate that the 80-kDa fragment is not an active tyrosine kinase. These results show that protein kinase C negatively regulates heregulin signaling through the ErbB-4 receptor by the activation of a selective proteolytic mechanism. PMID- 8702565 TI - Physical interaction between human RAD52 and RPA is required for homologous recombination in mammalian cells. AB - The yeast RAD52 protein is essential for DNA double-strand break repair, and meiotic and mitotic recombination. RPA is a protein complex of three subunits (70, 34, and 11 kDa) that has been shown to be involved in DNA replication, nucleotide excision repair, and homologous recombination. Here, we demonstrate a physical interaction between human RAD52 and RPA in vivo and in vitro. In addition, the domain (amino acids 221-280) in RAD52 protein that mediates the interaction with the 34-kDa subunit of RPA was also determined. Overexpression of mutant RAD52 proteins lacking the interaction domain (amino acids 221-240, 241 260, and 261-280) failed to induce homologous recombination in monkey cells. We have previously shown that overexpression of human RAD52 induced homologous recombination in these cells. These results suggest that direct physical interactions between RAD52 and RPA are essential for homologous recombination in mammalian cells. PMID- 8702566 TI - Domain analysis of human U5 RNA. Cap trimethylation, protein binding, and spliceosome assembly. AB - We have analyzed the sequence requirements of the human U5 RNA during small nuclear ribonucleoprotein (snRNP) and spliceosome assembly. A collection of mutant derivatives of the human U5 RNA gene was constructed in a U1 expression vector and transiently transfected in mammalian cells. Using immunoprecipitation and affinity selection assays, the cap trimethylation, the binding of Sm proteins and of the U5 snRNP-specific protein p220, as well as the assembly of the U4/U5/U6 triple snRNP and of spliceosomes were determined. By mutational analysis we were able to assign distinct functions to several structural elements of the human U5 RNA. Efficient binding of the Sm proteins requires the 3' stem-loop. Both the Sm protein-binding site and the 3' stem-loop are necessary for the formation of the trimethyl guanosine cap, consistent with Sm protein binding being a prerequisite for cap trimethylation. Specific elements of the U5 RNA 5' stem-loop contribute to efficient p220 association, in particular stem Ib. Interestingly, the highly conserved loop I appears to be a multifunctional element; in addition to its function in splice-site selection the 5' loop is involved in binding of p220 and in the assembly of the U4/U5/U6 triple snRNP. In sum, this mutational analysis has identified four functional domains of the human U5 RNA. PMID- 8702568 TI - The androgen-specific probasin response element 2 interacts differentially with androgen and glucocorticoid receptors. AB - The nuclear receptors constitute a large family of transcription factors characterized by a well conserved DNA-binding domain. The receptors for glucocorticoids, progestins, mineralocorticoids, and androgens constitute a subgroup because they bind in vitro with high affinity to DNA elements containing a partial palindrome of the core sequence 5'-TGTTCT-3'. In vivo, however, the corresponding steroids differentially regulate the expression of their target genes, even when more than one receptor type is present in a particular cell. The DNA-binding domains of the androgen and of the glucocorticoid receptors bind most androgen response elements with similar relative affinities. In contrast, one element (5'-GGTTCTTGGAGTACT-3') which was recently described in the promoter region of the probasin gene selectively interacts with the DNA-binding domain of the androgen receptor and not with that of the glucocorticoid receptor. From studies with chimeric elements, it can be deduced that it is the left subsequence 5'-GGTTCT-3' which excludes the glucocorticoid receptor domain from binding. In co-transfection experiments where the ARE of the C3(1) gene is responsive to both androgens and glucocorticoids, the probasin element is induced only by androgens and not by glucocorticoids. The existence of response elements which are recognized preferentially by the androgen receptor provides yet another possible mechanism to explain the differences of the in vivo effects between androgens and other steroids of the subgroup. PMID- 8702569 TI - The pleckstrin homology domain mediates transformation by oncogenic dbl through specific intracellular targeting. AB - The pleckstrin homology (PH) domain is an approximately 100 amino acid structural motif found in many cellular signaling molecules, including the Dbl oncoprotein and related, putative guanine nucleotide exchange factors (GEFs). Here we have examined the role of the Dbl PH (dPH) domain in the activities of oncogenic Dbl. We report that the dPH domain is not involved in the interaction of Dbl with small GTP-binding proteins and is incapable of transforming NIH 3T3 fibroblasts. On the other hand, co-expression of the dPH domain with oncogenic Dbl inhibits Dbl-induced transformation. A deletion mutant of Dbl that lacks a significant portion of the PH domain retains full GEF activity, but is completely inactive in transformation assays. Replacement of the PH domain by the membrane-targeting sequence of Ras is not sufficient for the recovery of transforming activity. However, subcellular fractionations of Dbl and Dbl mutants revealed that the PH domain is necessary and sufficient for the association of Dbl with the Triton X 100-insoluble cytoskeletal components. Thus, our results suggest that the dPH domain mediates cellular transformation by targeting the Dbl protein to specific cytoskeletal locations to activate Rho-type small GTP-binding proteins. PMID- 8702570 TI - Morphine activates opioid receptors without causing their rapid internalization. AB - We have examined the endocytic trafficking of epitope-tagged delta and mu opioid receptors expressed in human embryonic kidney (HEK) 293 cells. These receptors are activated by peptide agonists (enkephalins) as well as by the alkaloid agonist drugs etorphine and morphine. Enkephalins and etorphine cause opioid receptors to internalize rapidly (t1/2 approximately 6 min) by a mechanism similar to that utilized by a number of other classes of receptor, as indicated by localization of internalized opioid receptors in transferrin-containing endosomes and inhibition of opioid receptor internalization by hypertonic media. Remarkably, morphine does not stimulate the rapid internalization of either delta or mu opioid receptors, even at high concentrations that strongly inhibit adenylyl cyclase. These data indicate that agonist ligands, which have similar effects on receptor-mediated signaling, can have dramatically different effects on the intracellular trafficking of a G protein-coupled receptor. PMID- 8702571 TI - The mixed lineage kinase SPRK phosphorylates and activates the stress-activated protein kinase activator, SEK-1. AB - SPRK (also called PTK-1 and MLK-3), a member of the mixed lineage kinase subfamily of (Ser/Thr) protein kinases, encodes an amino-terminal SH3 domain followed by a kinase catalytic domain, two leucine zippers interrupted by a short spacer, a Rac/Cdc42 binding domain, and a long carboxyl-terminal proline-rich region. We report herein that SPRK activates the stress-activated protein kinases (SAPKs) but not ERK-1 during transient expression in COS cells; the p38 kinase is activated modestly (1.3-2 fold) but consistently. SPRK also activates cotransfected SEK-1/MKK-4, a dual specificity kinase which phosphorylates and activates SAPK. Reciprocally, expression of mutant, inactive SEK-1 inhibits completely the basal and SPRK-activated SAPK activity. Immunoprecipitated recombinant SPRK is able to phosphorylate and activate recombinant SEK-1 in vitro to an extent comparable to that achieved by MEK kinase-1. These results identify SPRK as a candidate upstream activator of the stress-activated protein kinases, acting through the phosphorylation and activation of SEK-1. PMID- 8702572 TI - Neu differentiation factor/neuregulin isoforms activate distinct receptor combinations. AB - The multiple isoforms of Neu differentiation factor (NDF/neuregulin) induce a pleiotropic cellular response that is isoform-specific and cell type-dependent. The molecular basis of this heterogeneity was addressed by comparing the two major groups of isoforms, alpha and beta. Both groups bind to the catalytically impaired receptor tyrosine kinase ErbB-3, whose mitogenic stimulation by NDF requires transactivation by other ErbB proteins, either ErbB-1 or ErbB-2. By expressing each pair of receptors in interleukin 3-dependent myeloid cells, we found that both isoforms induced mitogenic signals in cells co-expressing the combination of ErbB-3 with ErbB-2. However, only the beta isoform stimulated cells that expressed both ErbB-3 and ErbB-1, and neither isoform was active on cells expressing ErbB-3 alone. Both isoforms bind to all ErbB-3-expressing cells, albeit with different affinities, but the co-stimulatory mitogenic effect is correlated with the ability of each auxiliary receptor to transphosphorylate ErbB 3. These results imply that NDF isoforms differ in their ability to induce receptor heterodimers; whereas both types of isoforms signal through ErbB-3/ErbB 2 heterodimers, only beta isoforms are able to stabilize ErbB-3/ErbB-1 heterodimers. PMID- 8702573 TI - Primary alcohols modulate the activation of the G protein-coupled receptor rhodopsin by a lipid-mediated mechanism. AB - The visual pigment rhodopsin is a prototypical member of the G protein-coupled receptor superfamily. In this study, we have investigated the effect of a series of n-alcohols on the formation of metarhodopsin II (MII), the photoactivated conformation of rhodopsin, which binds and activates transducin. When rhodopsin was photolyzed in the presence of several n-alcohols, increased MII formation was observed in the order ethanol > butanol > hexanol, whereas longer chain n alcohols inhibited MII formation with decanol > octanol. The magnitude of the stimulatory effects was greater in a more highly unsaturated phospholipid. Alcohols, which enhanced MII formation also increased phospholipid acyl chain packing free volume, while those that decreased this bilayer property inhibited MII formation. An apparent discontinuity in the effect of these alcohols results when their potency is calculated in terms of the total aqueous alcohol concentration. In sharp contrast, a continuous variation in their behavior is observed, when their potency is calculated in terms of the amount of alcohol partitioned in the membrane. Our findings strongly support a lipid-mediated mechanism of action for alcohols on rhodopsin and, by analogy, for other G protein-coupled receptors. PMID- 8702574 TI - Modulation of human muscle sodium channels by intracellular fatty acids is dependent on the channel isoform. AB - Free fatty acids (FFAs), including arachidonic acid (AA), are implicated in the direct and indirect modulation of a spectrum of voltage-gated ion channels. Skeletal muscle sodium channels can be either activated or inhibited by FFA exposure; the response is dependent on both FFA structure and site of exposure. Recombinant human skeletal muscle sodium channels (hSkM1) were transfected into heterologous human renal epithelium HEK293t cells. Cytoplasmic delivery of 5 microM AA augmented the voltage-activated sodium current of hSkM1 channels by 190% (+/-54 S.E., n = 7) over a 20-min period. Similar results were seen with 5 microM oleic acid. Sodium currents in HEK293t cells transfected with human cardiac muscle sodium channels (hH1) were insensitive to AA treatment, and exposure to oleic acid inhibited the hH1 currents over a 20-min period by 29% (+/ 13 S.E., n = 5). The increase in hSkM1 current was not accompanied by shifts in voltage dependence of activation, steady-state inactivation, or markedly altered kinetics of inactivation of the macroscopic current. The FFA-induced increase in sodium currents was not dependent on protein kinase C activity. In contrast, both isoforms were reversibly inhibited by external application of unsaturated FFA. Thus, the differential effects of FFA on skeletal muscle sodium channels first noted in cultured muscle cells can be reproduced by expressing recombinant sodium channels in epithelial cells. Although the responses to applied FFAs could be direct or indirect, we suggest that: 1) SkM1 has two classes of response to FFA, one which produces augmentation of macroscopic currents with intracellular FFA, and a second which produces inhibition with extracellular FFA; 2) H1 has only one class of response, which produces inhibition with extracellular FFA. A testable hypothesis is that the presence or absence of each response is due to a specific structure in SkM1 or H1. These specific structures may directly interact with FFA or may interact with intermediate components. PMID- 8702575 TI - Tissue-specific isoforms of chicken myomesin are generated by alternative splicing. AB - Myomesin is a high molecular weight protein that is present in the M-band of all fiber types of cross-striated skeletal muscle and heart. We have isolated two cDNAs encoding tissue-specific isoforms of chicken myomesin with calculated molecular masses of 174 kDa in skeletal muscle and 182 kDa in heart. Distinct sequences are found at the 3'-end of the two cDNAs, giving rise to different C terminal domains. Partial analysis of the gene structure has shown that in chicken, both isoforms are generated by alternative splicing of a composite exon. Amino acid sequences show that the main body of myomesin consists of five fibronectin type III (class I motifs) and seven immunoglobulin-like domains (class II motifs). An identical structure was found in M-protein and human 190K protein (the human counterpart of chicken myomesin), and a comparable domain arrangement occurs in the M-band-associated protein skelemin. We postulate that myomesin, M-protein, and skelemin belong to the same subfamily of high molecular weight M-band-associated proteins of the immunoglobulin superfamily and that they probably have the same ancestor in evolution. PMID- 8702576 TI - Human apolipoprotein E4 domain interaction. Arginine 61 and glutamic acid 255 interact to direct the preference for very low density lipoproteins. AB - Human apolipoprotein (apo) E contains an amino- and a carboxyl-terminal domain, which are connected by a hinge region (approximately residues 165 to 215). The interaction of the two domains has been suggested to be responsible for the apoE4 binding preference for very low density lipoproteins (VLDL). In the absence of this interaction in apoE3, the preference is for high density lipoproteins (HDL). To exclude the possibility that the interaction of apoE with other apolipoproteins on the native particles may contribute to the isoform-specific preferences, VLDL-like emulsion particles were incubated with apoE, and the lipid bound apoE was separated from free apoE on a Superose 6 column. The apoE4 bound more effectively to these particles than did apoE3, indicating that the apoE4 preference for VLDL is due not to interactions with other apolipoproteins but to an intrinsic property of apoE4, likely related to domain interaction. Previously, arginine 61 was shown to be critical for the isoform preferences, suggesting that it interacted with an acidic residue(s) in the carboxyl terminus. Substitution of arginine 61 with lysine did not alter the preference of apoE4 for VLDL, demonstrating that a positive charge rather than a specific requirement for arginine is critical for domain interaction. To identify the acidic residue(s) in the carboxyl terminus interacting with arginine 61, the six acidic residues (244, 245, 255, 266, 270, and 271) in a region known to be important for both lipoprotein association and isoform-specific preferences were substituted individually with alanine in apoE4. Only substitution of glutamic acid 255 altered the preference of apoE4 from VLDL to HDL, indicating that this was the sole residue in the carboxyl terminus that interacts with arginine 61. The participation of the hinge region in domain interaction was examined with internal deletion mutants. Deletion of the residues 186-202 or 186-223, representing major portions of the hinge region, had no effect on the apoE4 preference for VLDL. This suggests that the hinge region may act as a spacer that connects the two domains. Further deletion into the carboxyl-terminal domain (to residue 244) results in a loss of apoE4 VLDL binding. These studies establish that interaction of arginine 61 and glutamic acid 255 mediates apoE4 domain interaction. PMID- 8702577 TI - Opposite effects of cholesteryl ester transfer protein and phospholipid transfer protein on the size distribution of plasma high density lipoproteins. Physiological relevance in alcoholic patients. AB - The aim of the present study was to investigate the role of the cholesteryl ester transfer protein (CETP) and the phospholipid transfer protein (PLTP) in determining the size distribution of high density lipoproteins (HDL) in human plasma. Whereas both purified CETP and PLTP preparations were able to promote the size redistribution of isolated HDL3, CETP favored the emergence of small HDL, while PLTP induced the formation of both small and large conversion products. When the total plasma lipoprotein fractions isolated from nine distinct subjects were incubated for 24 h at 37 degrees C with either purified PLTP or purified CETP, significant alterations in the relative proportions of the five distinct plasma HDL subpopulations, i.e., HDL2b (9.71-12.90 nm), HDL2a (8.77-9.71 nm), HDL3a (8.17-8.77 nm), HDL3b (7.76-8.17 nm), and HDL3c (7.21-7. 76 nm) were also observed. PLTP induced a significant increase in the relative abundance of HDL2b (8.66 +/- 2.34% versus 7.87 +/- 1. 83% in controls; p < 0.01) and a significant decrease in the relative abundance of HDL3a (32.76 +/- 3.42% versus 37.87 +/- 2.62% in controls; p < 0.05). In contrast, CETP significantly reduced the relative proportion of HDL2a (33.03 +/- 2.53% versus 37.56 +/- 6.43% in controls; p < 0.01) but significantly increased the relative proportion of both HDL3b (21.36 +/- 6.97% versus 15.58 +/- 7.75% in controls; p < 0.01) and HDL3c (3.21 +/ 4.84% versus 1.13 +/- 0.56% in controls; p < 0.05). Finally, in order to assess further the physiological relevance of in vitro observations, CETP activity, PLTP activity, and HDL size distribution were determined in plasmas from 33 alcoholic patients entering a cessation program. Alcohol withdrawal was associated with (i) a significant increase in plasma CETP activity (173.5 +/- 70.5%/h/ml before versus 223.2 +/- 69. 3%/h/ml after alcohol withdrawal, p = 0.0007), (ii) a significant reduction in plasma PLTP activity (473.9 +/- 203.7%/h/ml before versus 312.7 +/- 148.4%/h/ml after alcohol withdrawal, p = 0.0001), and (iii) a significant shift of large HDL2b and HDL2a toward small HDL3b and HDL3c. On the one hand, changes in plasma CETP activity correlated negatively with changes in the proportion of HDL2a (r = -0.597, p = 0.0002) and positively with changes in the proportion of HDL3b (r = 0.457, p = 0.0075). On the other hand, changes in plasma PLTP activity correlated positively with changes in the proportion of HDL2b (r = 0.482, p = 0.0045) and negatively with changes in the proportion of HDL3a (r = -0.418, p = 0.0154). Taken together, data of the present study revealed that plasma PLTP and CETP can exert opposite effects on the size distribution of plasma HDL. PLTP can promote the formation of HDL2b particles at the expense of HDL3a, while CETP can promote the formation of HDL3b particles at the expense of HDL2a. PMID- 8702578 TI - Stimulation of cardiac sarcoplasmic reticulum calcium pump by acylphosphatase. Relationship to phospholamban phosphorylation. AB - Ca2+ transport by cardiac sarcoplasmic reticulum is tightly coupled with the enzymatic activity of Ca2+-dependent ATPase, which forms and decomposes an intermediate phosphoenzyme. Heart sarcoplasmic reticulum Ca2+ pump is regulated by cAMP-dependent protein kinase (PKA) phospholamban phosphorylation, which results in a stimulation of the initial rates of Ca2+ transport and Ca2+ ATPase activity. In the present studies we found that acylphosphatase from heart muscle, used at concentrations within the physiological range, actively hydrolyzes the phosphoenzyme of cardiac sarcoplasmic reticulum Ca2+ pump, with an apparent Km on the order of 10(-7) M, suggesting an high affinity of the enzyme for this special substrate. In unphosphorylated vesicles acylphosphatase enhanced the rate of ATP hydrolysis and Ca2+ uptake with a concomitant significant decrease in apparent Km for Ca2+ and ATP. In vesicles whose phospholamban was PKA-phosphorylated, acylphosphatase also stimulated the rate of Ca2+ uptake and ATP hydrolysis but to a lesser extent, and the Km values for Ca2+ and ATP were not significantly different with respect to those found in the absence of acylphosphatase. These findings suggest that acylphosphatase, owing to its hydrolytic effect, accelerates the turnover of the phosphoenzyme intermediate with the consequence of an enhanced activity of Ca2+ pump. It is known that phosphorylation of phospholamban results in an increase of the rate at which the phosphoenzyme is decomposed. Thus, as discussed, a competition between phospholamban and acylphosphatase effect on the phosphoenzyme might be proposed to explain why the stimulation induced by this enzyme is less marked in PKA-phosphorylated than in unphosphorylated heart vesicles. PMID- 8702579 TI - Insulin exocytosis and glucose-mediated increase in cytoplasmic free Ca2+ concentration in the pancreatic beta-cell are independent of cyclic ADP-ribose. AB - Stimulation of pancreatic beta-cells by glucose gives rise to an increase in the cytoplasmic free calcium concentration ([Ca2+]i) and exocytosis of insulin. Cyclic adenosine 5'-diphosphate ribose (cADPR), a metabolite of beta-NAD+, has been reported to increase [Ca2+]i in pancreatic beta-cells by releasing Ca2+ from inositol 1,4,5-trisphosphate-insensitive intracellular stores. In the present study, we have examined the role of cADPR in glucose-mediated increases in [Ca2+]i and insulin exocytosis. Dispersed ob/ob mouse beta-cell aggregates were either pressure microinjected with fura-2 salt or loaded with fura-2 acetoxymethyl ester, and [Ca2+]i was monitored by microfluorimetry. Microinjection of beta-NAD+ into fura-2-loaded beta-cells did not increase [Ca2+]i nor did it alter the cells' subsequent [Ca2+]i response to glucose. Cells microinjected with the cADPR antagonist 8NH2-cADPR increased [Ca2+]i in response to glucose equally well as those injected with cADPR. Finally, the ability of cADPR to promote exocytosis of insulin in electropermeabilized beta-cells was investigated. cADPR on its own did not increase insulin secretion nor did it potentiate Ca2+-induced insulin secretion. We conclude that cADPR neither plays a significant role in glucose-mediated increases in [Ca2+]i nor interacts directly with the molecular mechanisms regulating exocytosis of insulin in normal pancreatic beta-cells. PMID- 8702580 TI - Changes in plasma lipoprotein cholesterol levels by antisense oligodeoxynucleotides against cholesteryl ester transfer protein in cholesterol fed rabbits. AB - Cholesteryl ester transfer protein (CETP) is the enzyme that facilitates the transfer of cholesteryl ester from high density lipoprotein (HDL) to apoB containing lipoproteins and also affects the low density lipoprotein metabolism. On the other hand, the liver is the major tissue responsible for the production of CETP (CETP mRNA) in rabbits. To test the hypothesis that a reduction of CETP mRNA in the liver by antisense oligodeoxynucleotides (ODNs) may affect the plasma lipoprotein cholesterol levels, we intravenously injected antisense ODNs against rabbit CETP coupled with asialoglycoprotein carrier molecules, which serve as an important method to regulate liver gene expression, to cholesterol-fed rabbits via their ear veins. All rabbits were fed a standard rabbit chow supplement with 0.1% cholesterol for 10 weeks before and throughout the experiment. After injecting rabbits with antisense ODNs, the plasma total cholesterol concentrations and plasma CETP activities all decreased at 24, 48, and 96 h, whereas the plasma HDL cholesterol concentrations increased at 48 h. A reduction in the hepatic CETP mRNA was also observed at 6, 24, and 48 h after the injection with antisense ODNs. However, in the rabbits injected with sense ODNs, the plasma total and HDL cholesterol concentrations and the plasma CETP activities did not significantly change, and the hepatic CETP mRNA did not change either throughout the experimental period. Although the exact role of CETP in the development of atherosclerosis remains to be clarified, these findings showed for the first time that the intravenous injection with antisense ODNs against CETP coupled to asialoglycoprotein carrier molecules targeted to the liver could thus inhibit plasma CETP activity and, as a result, could induce a decrease in the plasma low density lipoprotein and very low density lipoprotein cholesterol and an increase in the plasma HDL cholesterol in cholesterol-fed rabbits. PMID- 8702581 TI - The amino-terminal extracellular domain of the MCP-1 receptor, but not the RANTES/MIP-1alpha receptor, confers chemokine selectivity. Evidence for a two step mechanism for MCP-1 receptor activation. AB - The chemoattractant cytokines, MCP-1 (monocyte chemoattractant protein) and MIP 1alpha (macrophage inflammatory protein), are recognized by highly homologous but distinct receptors. To identify receptor domains involved in determining ligand specificity, we created a series of chimeric MCP-1 and RANTES (regulated on activation, normal T cell expressed and secreted)/MIP-1alpha receptors that progressively interchanged the amino terminus and each of the three extracellular loops. Radiolabeled MCP-1 bound with high affinity to the wild-type MCP-1 receptor, but not to the RANTES/MIP-1alpha receptor (C-C CKR-1). Chimeras that retained the amino-terminal extension of the MCP-1 receptor bound MCP-1 with high affinity. In contrast, chimeric MCP-1 receptors, in which the wild-type amino terminus was replaced with the corresponding portion of the RANTES/MIP-1alpha receptor, bound MCP-1 with low affinity. These data indicate that the amino terminus of the MCP-1 receptor is necessary for high affinity binding of the ligand. Very different results were obtained using the RANTES/MIP-1alpha receptor. Radiolabeled MIP-1alpha bound with high affinity to chimeras that expressed the extracellular loops of the RANTES/MIP-1alpha receptor. In contrast to the MCP-1 receptor, substitution of the wild-type amino-terminal extension had little or no effect on MIP-1alpha binding. For the MCP-1, but not the RANTES/MIP 1alpha receptor, the presence of the wild-type amino terminus also significantly lowered the ligand concentration required for maximal signaling. We conclude that the amino-terminal extension of the MCP-1 receptor, but not the RANTES/MIP-1alpha receptor, is critically involved in ligand binding and signal transduction. These data reveal significant functional differences between the two C-C chemokine receptors and suggest a two-step mechanism for activation of the MCP-1 receptor. PMID- 8702582 TI - Expression of Kv1.1 delayed rectifier potassium channels in Lec mutant Chinese hamster ovary cell lines reveals a role for sialidation in channel function. AB - Kv1.1 potassium (K+) channels contain significant amounts of negatively charged sialic acids. To examine the role of sialidation in K+ channel function, Chinese hamster ovary cell lines deficient in glycosylation (Lec mutants) were transfected with rat brain Kv1.1 cDNA. The K+ channel was functionally expressed in all cell lines, but the voltage dependence of activation (V1/2) was shifted to more positive voltages and the activation kinetics were slower in the mutant cell lines compared with control. A similar positive shift in V1/2 was recorded in control cells expressing Kv1.1 following treatment with sialidase or by raising extracellular Ca2+. In contrast, these treatments had little or no effect on the Lec mutants, which indicates that channel sialic acids appear to be the negative surface charges sensitive to Ca2+. The data suggest that sialic acid addition modifies Kv1.1 channel function, possibly by influencing the local electric field detected by its voltage sensor, but that these carbohydrates are not required for cell surface expression. PMID- 8702583 TI - The levels of ribonucleotide reductase, thioredoxin, glutaredoxin 1, and GSH are balanced in Escherichia coli K12. AB - The dithiol forms of thioredoxin and glutaredoxin are hydrogen donors for ribonucleotide reductase. We have determined the intracellular levels of ribonucleotide reductase (RRase), thioredoxin (Trx), glutaredoxin 1 (Grx1), and glutathione (GSH) and the glutathione redox status in new Escherichia coli K12 strains lacking thioredoxin (trxA-), glutaredoxin 1 (grxA-), and/or GSH (gshA-) or overproducing Trx or Grx1 from multicopy plasmids. We propose a regulatory network in which RRase levels are balanced with those of Trx, Grx1, and GSH so that deficiency or overproduction of one component would promote the opposite effect on the others to maintain a balanced supply of deoxyribonucleotides. GSH deficiency strongly increased both Grx1 levels and RRase activity, even more than Trx deficiency. Double gshA-trxA- bacteria were viable, whereas additional deficiency in lipoate synthesis (gshA-trxA-lipA-) caused the inability to grow in minimal medium plates supplemented with acetate plus succinate instead of lipoic acid. Thus, lipoate might be the only substitute of GSH for glutaredoxin reduction in gshA-trxA- cells, although the extremely high Grx1 content (55-fold) of these bacteria suggests that electron transfer from lipoate might be an inefficient reduction mechanism of glutaredoxins. Moreover, the enhanced Grx1 level of gshA-trxA- cells could obviate the need for a large increase in RRase activity, in contrast to grxA-trxA- double mutant cells. Impairment of the sulfate assimilation pathway, leading to very low GSH concentrations, and an oxidized glutathione redox state might explain the inability of grxA-trxA- cells to grow in minimal medium. Restoration of nearly normal levels of both GSH content and redox status cure the growth defect. PMID- 8702584 TI - Acylation of pulmonary surfactant protein-C is required for its optimal surface active interactions with phospholipids. AB - This study investigates the importance of thioester-linked acyl groups in lung surfactant protein C (SP-C) in facilitating interactions with phospholipids that yield functionally important surface active behaviors. Native SP-C, palmitoylated at cysteine residues at positions 5 and 6, was isolated from bovine lung surfactant by liquid chromatography. Deacylated SP-C (dSP-C), unchanged in composition and sequence from SP-C but having a decreased alpha-helical content in films with dipalmitoyl phosphatidylcholine (DPPC) of 52 versus 70%, was obtained by treatment with 0.1 M sodium carbonate buffer at pH 10. Surface activity was studied for SP-C and dSP-C combined with column-purified phospholipids (PPL) from calf lung surfactant or with synthetic phospholipids (DPPC or a synthetic phospholipid mixture (SPL) containing 50:35:15, DPPC:egg phosphatidylcholine:egg phosphatidylglycerol). Interfacial measurements included surface pressure time adsorption isotherms for dispersed surfactants with diffusion minimized, dynamic surface pressure area isotherms and respreading for films in the Wilhelmy balance, and overall surface tension lowering at physiologic cycling rate in oscillating bubble experiments. Dispersions of PPL:SP C and SPL:SP-C rapidly adsorbed to high equilibrium surface pressures of 47-48 mN/m, significantly better than corresponding dispersions containing dSP-C. The adsorption of PPL:dSP-C was essentially unchanged from that of PPL alone, and the adsorption of SPL:dSP-C was improved only slightly over SPL alone. In Wilhelmy balance studies, dynamic respreading was significantly improved over phospholipids alone in films of SP-C plus PPL, SPL, or DPPC. Respreading was improved less markedly by dSP-C in corresponding films with SPL or DPPC and not at all in films with PPL. Maximum surface pressures were also higher in cycled films of SP-C versus dSP-C combined with PPL or SPL. In bubble experiments (37 degrees C, 20 cycles/min), dispersions of PPL:SP-C and SPL:SP-C reached low minimum surface tensions of <1 and 5 mN/m, respectively, whereas PPL:dSP-C and SPL:dSP-C only reached minima of approximately 20 mN/m as did PPL and SPL alone. Acylation in SP-C is crucial for its interactions with phospholipids over the full spectrum of adsorption and dynamic surface behaviors important for lung surfactant. PMID- 8702585 TI - Mutations in a specific human serum albumin thyroxine binding site define the structural basis of familial dysalbuminemic hyperthyroxinemia. AB - The familial dysalbuminemic hyperthyroxinemia (FDH) phenotype results from a natural human serum albumin (HSA) mutant with histidine instead of arginine at amino acid position 218. This mutation results in an enhanced affinity for thyroxine. Site-directed mutagenesis and a yeast protein expression system were used to synthesize wild type HSA and FDH HSA as well as several other HSA mutants. Studies on the binding of thyroxine to these HSA species using equilibrium dialysis and quenching of tryptophan 214 fluorescence suggest that the FDH mutation affects a single thyroxine binding site located in the 2A subdomain of HSA. Site-directed mutagenesis of HSA and thyroxine analogs were used to obtain information about the mechanism of thyroxine binding to both wild type and FDH HSA. These studies suggest that the guanidino group of arginine at amino acid position 218 in wild type HSA is involved in an unfavorable binding interaction with the amino group of thyroxine, whereas histidine at amino acid position 218 in FDH HSA is involved in a favorable binding interaction with thyroxine. Neither arginine at amino acid position 222 nor tryptophan at amino acid position 214 appears to favorably influence the binding of thyroxine to wild type HSA. PMID- 8702586 TI - Assembly of the B subunit pentamer of Escherichia coli heat-labile enterotoxin. Kinetics and molecular basis of rate-limiting steps in vitro. AB - The B subunits of Escherichia coli heat-labile enterotoxin (EtxB) and cholera toxin (CtxB) assemble in vivo into exceptionally stable homopentameric complexes, which maintain their quaternary structure in a range of conditions that would normally be expected to cause protein denaturation. Recently, we showed that the simultaneous protonation of two of the COOH-terminal carboxylates in pentameric EtxB was required to cause its disassembly at pH values below 2.0 (Ruddock, L., Ruston, S. P., Kelly, S. M., Price, N. C., Freedman, R. B., and Hirst, T. R.(1995) J. Biol. Chem. 270, 29953-29958). Here, we investigate the influence of environmental parameters on the kinetics of reassembly of acid-generated EtxB monomers in vitro. Such monomers were found to undergo a further acid-mediated conformational change, with an activation energy of 76 +/- 2 J.mol-1.K-1, consistent with isomerization of the cis-proline residue at position 93, and which prevented subsequent EtxB reassembly. By using rapid neutralization of acid generated monomers, a high proportion of the B-subunits adopted an assembly competent conformation, which resulted in up to 75% of the protein reassembling into a stable pentameric complex, indistinguishable from native EtxB pentamers. The rate-limiting step in reassembly, over a concentration range of 50-200 microg/ml, was shown to be due to an intramolecular event, which exhibited a pH dependence with a pKa of 7.0. Modification of EtxB with amine-specific probes revealed that the protonation state of the NH2-terminal alanine residue was responsible for the pH dependence of reassembly. The implications of these findings for the biogenesis of Escherichia coli enterotoxin and related enterotoxins in vivo, are considered. PMID- 8702587 TI - Isocitrate dehydrogenase kinase/phosphatase. Kinetic characteristics of the wild type and two mutant proteins. AB - Isocitrate dehydrogenase (IDH) of Escherichia coli is regulated by a bifunctional protein, IDH kinase/phosphatase. In addition to the kinase and phosphatase activities, this protein catalyzes an intrinsic ATPase reaction. The initial velocity kinetics of these activities exhibited extensive similarities. IDH kinase and phosphatase both yielded intersecting double-reciprocal plots. In addition, we observed similar values for the kinetic constants describing interactions of the kinase and phosphatase with their protein substrates and the interactions of all three activities with ATP. In contrast, while the maximum velocities of IDH kinase and IDH phosphatase were nearly equal, they were 10-fold less than the maximum velocity of the ATPase. Although the IDH phosphatase reaction required either ATP or ADP, it was not supported by the nonhydrolyzable ATP analogue 5'-adenylyl imidodiphosphate. The kinetic properties of wild-type IDH kinase/phosphatase were compared with those of two mutant derivatives of this protein. The mutations in these proteins selectively inhibit IDH phosphatase activity. Inhibition of IDH phosphatase resulted from three factors: decreases in the maximum velocities, reduced affinities for phospho-IDH, and a loss of coupling between ATP and phospho-IDH. These mutations also affected the properties of IDH kinase, increasing the maximum velocities and decreasing the affinities for ATP and phospho-IDH. The intrinsic ATPase activities also exhibited reduced affinity for ATP. These results are discussed in the context of a model which proposes that all three activities occur at the same active site. PMID- 8702588 TI - Role of oxoproline in the regulation of neutral amino acid transport across the blood-brain barrier. AB - Regulation of neutral amino acid transport was studied using isolated plasma membrane vesicles derived from the bovine blood-brain barrier. Neutral amino acids cross the blood-brain barrier by facilitative transport system L1, which may allow both desirable and undesirable amino acids to enter the brain. The sodium-dependent amino acid systems A and Bo,+ are located exclusively on abluminal membranes, in a position to pump unwanted amino acids out. gamma Glutamyl transpeptidase, the first enzyme of the gamma-glutamyl cycle, is an integral protein of the luminal membrane of the blood-brain barrier. We demonstrate that oxoproline, an intracellular product of the gamma-glutamyl cycle, stimulates the sodium-dependent systems A and Bo,+ by 70 and 20%, respectively. Study of system A showed that 2 mM oxoproline increased the affinity for its specific substrate N-methylaminoisobutyrate by 50%. This relationship between the activity of the gamma-glutamyl cycle and system A transport may provide a short term regulatory mechanism by which the entry of potentially deleterious amino acids (i.e. neurotransmitters or their precursors) may be retarded and their removal from brain accelerated. PMID- 8702589 TI - Role of Val509 in time-dependent inhibition of human prostaglandin H synthase-2 cyclooxygenase activity by isoform-selective agents. AB - Prostaglandin H synthase (PGHS), a key enzyme in prostanoid biosynthesis, exists as two isoforms. PGHS-1 is considered a basal enzyme; PGHS-2 is associated with inflammation and cell proliferation. A number of highly selective inhibitors for PGHS-2 cyclooxygenase activity are known. Inhibition by these agents involves an initial reversible binding, followed by a time-dependent transition to a much higher affinity enzyme-inhibitor complex, making these agents potent and poorly reversible PGHS-2 inhibitors. To investigate the PGHS-2 structural features that influence the time-dependent action of the selective inhibitors, we have constructed a three-dimensional model of human PGHS-2 by homologous modeling. Examination of the PGHS-2 model identified Val509 as a cyclooxygenase active site residue, that was not conserved in PGHS-1. Recombinant human PGHS-2 with Val509 mutated to either Ile (the corresponding residue in PGHS-1), Ala, Glu, or Lys was expressed by transient transfection of COS-1 cells to evaluate the effects of the mutations on cyclooxygenase activity and on inhibition by four agents reported to be selective for PGHS-2 (NS398, nimesulide, DuP697, and SC58125). All the recombinant proteins were of the expected mass. The mutants exhibited 45-210% of wild-type cyclooxygenase activity, with Km values for arachidonate of 2.1-7.6 microM (wild-type PGHS-2, 3.8 microM), indicating that changes in position 509 had modest effects on cyclooxygenase catalysis. Each of the agents inhibited wild type PGHS-2 in a time-dependent fashion, and all but nimesulide did the same for the V509A mutant. In contrast, the V509E and V509I PGHS-2 mutants, like recombinant human PGHS-1, did not show time-dependent inhibition with any of the agents, and the V509K mutant responded in a time-dependent manner only to DuP697. Reversible inhibition was still observed with Val509 mutants that did not show time-dependent inhibition. Thus, the side chain structure at position 509 markedly influenced the ability of PGHS-2 to undergo the time-dependent transition without removing inhibitor or substrate binding. These results indicate that Val509 in PGHS-2 has a major role in the structural transition that underlies time-dependent inhibition by the isoform-selective agents. PMID- 8702590 TI - Importance of the two interferon-stimulated response element (ISRE) sequences in the regulation of the human indoleamine 2,3-dioxygenase gene. AB - Indoleamine 2,3-dioxygenase (INDO) is the rate-limiting enzyme in the catabolism of the essential amino acid L-tryptophan. It is induced strongly in many cell lines following interferon-gamma treatment. We report the cloning and characterization of the full-length human INDO promoter. This promoter is 1,245 base pairs long and includes two interferon-stimulated response elements (ISRE) separated by an approximately 1-kilobase sequence. The presence of these two ISREs is critical for maximum INDO promoter activity (50-fold induction). When the ISREs are present in two separate fragments cloned upstream of the chloramphenicol acetyltransferase (CAT) reporter vector, the INDO promoter activity drops significantly (7-fold induction). 5' end deletions of the wild type promoter sequence indicate that removal of the ISRE (ISRE1) at position 1126 reduces the induction level to approximately 25-fold. This activity does not change appreciably when the promoter is deleted down to position -241. Furthermore, site-directed mutagenesis of ISRE1 also decreases the promoter activity in a similar way. When ISRE1 is kept intact, deletion of the second ISRE (ISRE2) at position -111 leads to only 11-fold induction of the promoter. A similar result is obtained when substitution mutations are introduced in ISRE2. Deletion of a 748-base pair sequence between the two ISREs only shows a slight decrease in the INDO promoter activity. These data indicate that the two ISRE sequences are required for the full transcriptional induction of the interferon gamma-inducible human INDO gene. INDO activity is not induced in the hepatic cell line HepG2. An analysis of INDO-CAT activity in this cell line indicated that the lack of INDO activity was at the transcriptional level and could reflect either the presence of a repressor or lack of a transcription factor. This lack of induction could be correlated with a truncated or unstable IRF-1. However, the levels of IRF-2, JAK2, and STAT 91 were similar in both ME180 and HepG2 cells. PMID- 8702591 TI - Phosphatidylinositol 3-kinase inhibitors block differentiation of skeletal muscle cells. AB - Skeletal muscle differentiation involves myoblast alignment, elongation, and fusion into multinucleate myotubes, together with the induction of regulatory and structural muscle-specific genes. Here we show that two phosphatidylinositol 3 kinase inhibitors, LY294002 and wortmannin, blocked an essential step in the differentiation of two skeletal muscle cell models. Both inhibitors abolished the capacity of L6E9 myoblasts to form myotubes, without affecting myoblast proliferation, elongation, or alignment. Myogenic events like the induction of myogenin and of glucose carrier GLUT4 were also blocked and myoblasts could not exit the cell cycle, as measured by the lack of mRNA induction of p21 cyclin dependent kinase inhibitor. Overexpresssion of MyoD in 10T1/2 cells was not sufficient to bypass the myogenic differentiation blockade by LY294002. Upon serum withdrawal, 10T1/2-MyoD cells formed myotubes and showed increased levels of myogenin and p21. In contrast, LY294002-treated cells exhibited none of these myogenic characteristics and maintained high levels of Id, a negative regulator of myogenesis. These data indicate that whereas phosphatidylinositol 3-kinase is not indispensable for cell proliferation or in the initial events of myoblast differentiation, i.e. elongation and alignment, it appears to be essential for terminal differentiation of muscle cells. PMID- 8702592 TI - Sphingomyelin inhibits the lecithin-cholesterol acyltransferase reaction with reconstituted high density lipoproteins by decreasing enzyme binding. AB - Lecithin-cholesterol acyltransferase (LCAT) catalyzes the formation of cholesterol esters on high density lipoproteins (HDL) and plays a critical role in reverse cholesterol transport. Sphingomyelin, an important constituent of HDL, may regulate the activity of LCAT at any of the key steps of the enzymatic reaction: binding of LCAT to the interface, activation by apo A-I, or inhibition at the catalytic site. In order to clarify the role of sphingomyelin in the regulation of the LCAT reaction and its effects on the structure of apolipoprotein A-I, we prepared reconstituted HDL (rHDL) containing egg phosphatidylcholine, cholesterol, apolipoprotein A-I, and up to 22 mol % sphingomyelin. Because the interfacial properties of substrate particles can dramatically affect LCAT binding and kinetics, we also prepared and analyzed proteoliposome substrates having the same components as the rHDL, except for a 4 fold higher ratio of phospholipid to apolipoprotein A-I. The reaction kinetics of LCAT with the rHDL particles revealed no significant change in the apparent Vmax but showed a concentration-dependent increase in slope of the reciprocal plots and in the apparent Km values with sphingomyelin content. The dissociation constant (Kd) for LCAT with these particles increased linearly with sphingomyelin content up to 22 mol %, changing in parallel with the apparent Km values. No structural changes of apolipoprotein A-I were detected in the particles with increasing content of sphingomyelin, but fluorescence results with lipophilic probes revealed that significant changes in the acyl chain, backbone, and head group regions of the lipid bilayer of the particles are introduced by the addition of sphingomyelin. On the other hand, the proteoliposome substrates also had increased Kdvalues for LCAT at high sphingomyelin contents but compared with the rHDL particles had a 6-10-fold lower affinity for LCAT binding and exhibited kinetics consistent with competitive inhibition by sphingomyelin at the active site. These results show conclusively that the dominant mechanism for the inhibition of LCAT activity with rHDL particles by sphingomyelin is the impaired binding of the enzyme to the interface. The results also underscore the significant differences in the enzyme reaction kinetics with different substrate particles. PMID- 8702593 TI - Unusual beta-D-xylosides that prime glycosaminoglycans in animal cells. AB - The biosynthesis of glycosaminoglycans (GAG) takes place while the polysaccharide chains are usually attached to a proteoglycan core protein. Cells also will assemble GAG chains on beta-D-xylosides containing hydrophobic aglycones. In order to evaluate the relationship of the structure of the sugar to priming activity of the glycoside, we synthesized beta-D-xyloside analogs in which the hydroxyls were substituted with hydrogen, fluorine, -O-methyl, amino, -O isopropyl, and -O-benzyl groups. Epimers at the 2-, 3-, and 4-position of xylose also were made. Their ability to prime GAGs was tested in Chinese hamster ovary cells by measuring 35SO4 incorporation into polysaccharide chains and by assaying the transfer of galactose to the xylosides by galactosyltransferase I (UDP-D galactose:xylose beta1-4-galactosyltransferase) in vitro. All of the analogs failed to act as primers of GAGs in vivo and as substrates in vitro with the following exceptions. Substitution of 2-OH and 3-OH with -OCH3 were active at high concentration (1 mM), but the deoxygenated derivatives were inactive. Efficient priming also occurred on a derivative with fluorine instead of the 3-OH group, suggesting that the oxygen atoms at C-2 and C-3 were involved as hydrogen bond acceptors. Methylated and deoxy analogs at C-4 were inactive, due to the loss of the acceptor hydroxyl group. Interestingly, benzyl-beta-D-threo pentopyranos-4-uloside (4-keto derivative) and benzyl-4-methyl-beta-D-xyloside, with a methyl group in place of an axial hydrogen at C-4, primed GAG chains. Priming by these unusual xylosides suggests the possibility of designing inhibitors of GAG synthesis based on xyloside analogs with reactive groups in key positions. PMID- 8702594 TI - Functional relationships of the genetic locus encoding the glycosyltransferase enzymes involved in expression of the lacto-N-neotetraose terminal lipopolysaccharide structure in Neisseria meningitidis. AB - The biosynthetic function of the lgtABE genetic locus of Neisseria meningitidis was determined by structural analysis of lipopolysaccharide (LPS) derived from mutant strains and enzymic assay for glycosyltransferase activity. LPS was obtained from mutants generated by insertion of antibiotic resistance cassets in each of the three genes lgtA, lgtB, lgtE of the N. meningitidis immunotype L3 strain phi3 MC58. LPS from the parent strain expresses the terminal lacto-N neotetraose structure, Galbeta1-->4GlcNAcbeta1-->3Galbeta1-->4Glc. Mild hydrazine treatment of the LPS afforded O-deacylated samples that were analyzed directly by electrospray ionization mass spectrometry (ESI-MS) in the negative ion mode. In conjunction with results from sugar analysis, ESI-MS revealed successive loss of the sugars Gal, GlcNAc, and Gal in lgt B, lgt A, and lgt E LPS, respectively. The structure of a sample of O- and N-deacylated LPS derived by aqueous KOH treatment of lgt B LPS was determined in detail by two-dimensional homo- and heteronuclear NMR methods. Using a synthetic beta-GlcNAc acceptor and a beta-lactose acceptor, the glycosyltransferase activities encoded by the lgtB and lgtA genes were unambiguously established. These data provide the first definitive evidence that the three genes encode the respective glycosyltransferases required for biosynthesis of the terminal trisaccharide moiety of the lacto-N-neotetraose structure in Neisseria LPS. From ESI-MS data, it was also determined that the Gal deficient LPS expressed by the lgt E mutant is identical to that of the major component expressed by immunotype L3 galE-deficient strains. The galE gene which encodes for UDP-glucose-4-epimerase plays an essential role in the incorporation of Gal into meningococcal LPS. PMID- 8702595 TI - Structural organization of the reduced folate carrier gene in Chinese hamster ovary cells. AB - The reduced folate carrier gene (rfc) encodes a putative protein that is involved in the intracellular accumulation of folates. In this report, we describe the organization of the rfc gene from Chinese hamster ovary cells. The hamster rfc gene contains 7 exons and 6 introns, which span 15.3 kilobases. It codes for two alternatively spliced messenger RNAs, one that contains all 7 exons and one that lacks exon 2 but contains the remaining 6 exons. The transcriptional start of the gene has been mapped to six sites approximately 200 base pairs upstream of the putative ATG initiation codon. The promoter region has no TATA box-like sequence but contains a consensus Sp1 binding site. This is the first report of the genomic structure of the reduced folate carrier gene from any species. PMID- 8702596 TI - S-nitrosoglutathione is cleaved by the thioredoxin system with liberation of glutathione and redox regulating nitric oxide. AB - In activated human neutrophils a burst of nitric oxide (NO) converts intracellular GSH to S-nitrosoglutathione (GSNO) which is subsequently cleaved to restore GSH by an unknown mechanism. We discovered that GSNO is an NADPH oxidizing substrate for human or calf thymus thioredoxin reductase (TR) with an apparent Km value of 60 microM and a Kcat of 0.6 x s-1. Addition of human thioredoxin (Trx) stimulated the initial NADPH oxidation rate severalfold but was accompanied by progressive inactivation of TR. Escherichia coli TR lacked activity with GSNO, but with E. coli Trx present, GSNO was reduced without inhibition of the enzyme. Chemically reduced E. coli Trx-(SH)2 was oxidized to Trx-S2 by GSNO with a rate constant of 760 M-1s-1 (7-fold faster than by GSSG) as measured by tryptophan fluorescence. Analysis of this reaction in the presence of oxymyoglobin revealed quantitative formation of metmyoglobin indicative of NO. release. Analysis of GSNO reduction demonstrated that oxidation of NADPH produced a stoichiometric amount of free GSH. These results demonstrate a homolytic cleavage mechanism of GSNO, giving rise to GSH and NO.. GSNO efficiently inhibited the protein disulfide reductase activity of the complete human or calf thymus thioredoxin systems. Our results demonstrate enzymatic cleavage of GSNO by TR or Trx and suggest novel mechanisms for redox signaling. PMID- 8702597 TI - Role of the C terminus of the interleukin 8 receptor in signal transduction and internalization. AB - Interleukin 8 (IL-8) is a potent neutrophil chemoattractant and activator. Two IL 8 receptor subtypes, A and B, are expressed in neutrophils. In this work, we analyzed the role of the C terminus domain of the IL-8 receptor on the signal transduction and receptor internalization mechanisms. The IL-8 receptor A was tagged with an epitope corresponding to the monoclonal antibody 1D4 to monitor the localization of the IL-8 receptor. We demonstrated IL-8-dependent receptor internalization by monitoring the density of surface 125I-labeled IL-8 binding sites and by immunofluorescence microscopy. Truncation of the last 27 amino acids of the IL-8 receptor A severely impaired the IL-8-induced internalization of the receptor. Of importance was the observation that binding of IL-8 to receptors A and B triggered a dramatically faster rate of internalization of receptor B than receptor A, suggesting that the heterologous C termini among receptor subtypes modulate the rate of internalization of IL-8 receptors. However, substitution of the C terminus of the receptor subtype A for the C terminus of receptor B reduced the internalization rate of receptor A. Furthermore, we found that the rate of internalization of IL-8 receptor B triggered by IL-8 was faster than the one induced by the IL-8-related peptide, melanoma growth stimulatory activity. Studies with human neutrophils pretreated with 100 nM IL-8 for 5 min revealed a positive and a negative calcium response mediated by receptors A and B, respectively. In contrast, neutrophils pretreated with melanoma growth stimulatory activity showed positive calcium responses to both receptors A and B. These data suggest that the neutrophil responses mediated by IL-8 are modulated by the rate of internalization of receptors. PMID- 8702598 TI - Rat brain contains high levels of mannose-6-phosphorylated glycoproteins including lysosomal enzymes and palmitoyl-protein thioesterase, an enzyme implicated in infantile neuronal lipofuscinosis. AB - Mannose 6-phosphate (Man-6-P) is a posttranslational carbohydrate modification typical of newly synthesized acid hydrolases that signals targeting from the Golgi apparatus to the lysosome via Man-6-P receptors (MPRs). Using iodinated cation independent MPR as a probe in a Western blot assay, we surveyed levels of Man-6-P glycoproteins in a number of different rat tissues. Considerable variation was observed with respect to total amounts and types of Man-6-P glycoproteins in the different tissues. Brain contained 2-8-fold more Man-6-P glycoproteins than other tissues, with relative abundance being brain >> testis approximately heart > lung approximately kidney approximately ovary approximately spleen > skeletal muscle approximately liver approximately serum. Analysis of 16 different lysosomal enzyme activities revealed that brain contains lower activities than other tissues which suggested that decreased removal of Man-6-P results in increased levels of Man-6-P glycoproteins. This was directly demonstrated by comparing activities of phosphorylated lysosomal enzymes, purified by immobilized MPR affinity chromatography, with total activities. The phosphorylated forms accounted for a considerable proportion of the MPR-targeted activities measured in brain (on average, 36.2%) but very little in lung, kidney, and liver (on average, 5.5, 2.3, and 0. 7%, respectively). Man-6-P glycoproteins were also isolated from rat brain by MPR affinity chromatography on a preparative scale. Of the 18 bands resolvable by SDS-polyacrylamide gel electrophoresis, seven bands were NH2-terminally sequenced and identified as the known lysosomal enzymes cathepsin L, cathepsin A, cathepsin D, alpha-galactosidase A, arylsulfatase A, and alpha-iduronidase. One of the major Man-6-P glycoproteins was identified as palmitoyl protein thioesterase, which was not previously thought to be lysosomal. This finding raises important questions about the cellular location and function of palmitoyl protein thioesterase, mutations in which result in the neurodegenerative disorder, infantile neuronal ceroid lipofuscinosis. PMID- 8702599 TI - Formation of nitrating and chlorinating species by reaction of nitrite with hypochlorous acid. A novel mechanism for nitric oxide-mediated protein modification. AB - Detection of 3-nitrotyrosine has served as an in vivo marker for the production of the cytotoxic species peroxynitrite (ONOO-). We show here that reaction of nitrite (NO2-), the autoxidation product of nitric oxide (.NO), with hypochlorous acid (HOCl) forms reactive intermediate species that are also capable of nitrating phenolic substrates such as tyrosine and 4-hydroxyphenylacetic acid, with maximum yields obtained at physiological pH. Monitoring the reaction of NO2- with HOCl by continuous flow photodiode array spectrophotometry indicates the formation of a transient species with spectral characteristics similar to those of nitryl chloride (Cl-NO2). Reaction of synthetic Cl-NO2 with N-acetyl-L tyrosine results in the formation of 3-chlorotyrosine and 3-nitrotyrosine in ratios that are similar to those obtained by the NO2-/HOCl reaction (4:1). Tyrosine residues in bovine serum albumin are also nitrated and chlorinated by NO2-/HOCl and synthetic Cl-NO2. The reaction of N-acetyl-L-tyrosine with NO2 /HOCl or authentic Cl-NO2 also produces dityrosine, suggesting that free radical intermediates are involved in the reaction mechanism. Our data indicate that while chlorination reactions of Cl-NO2 are mediated by direct electrophilic addition to the aromatic ring, a free radical mechanism appears to be operative in nitrations mediated by NO2-/HOCl or Cl-NO2, probably involving the combination of nitrogen dioxide (.NO2) and tyrosyl radical. We propose that NO2- reacts with HOCl by Cl+ transfer to form both cis- and trans-chlorine nitrite (Cl-ONO) and Cl NO2 as intermediates that modify tyrosine by either direct reaction or after decomposition to reactive free and solvent-caged Cl. and .NO2 as reactive species. Formation of Cl-NO2 and/or Cl-ONO in vivo may represent previously unrecognized mediators of inflammation-mediated protein modification and tissue injury, and offers an additional mechanism of tyrosine nitration independent of ONOO-. PMID- 8702600 TI - Presence of a light-independent phospholipase A2 in bovine retina but not in rod outer segments. AB - Rod outer segments (ROS) are responsible for the visual transduction process. Rhodopsin, which constitutes 85-90% of ROS proteins, absorbs light photons, changes its conformation, and then binds to a heterotrimeric G-protein called transducin. As a consequence, transducin dissociates into Talpha and Tbetagamma subunits. The presence in ROS of a phospholipase A2 (PLA2) stimulated by light and guanosine 5'-O-(3-thio)triphosphate was first demonstrated in 1987 (Jelsema, C. L.(1987) J. Biol. Chem. 262, 163-168). This led that author to conclude that ROS PLA2 could be involved in the phototransduction process, and raised the possibility of receptor-mediated activation of PLA2 via G-proteins in cell types other than rods. However, the biochemical characteristics and the role of this PLA2 have not been fully elucidated. We have tried to reproduce some of the results previously reported in order to further characterize this enzyme. We have found that, in our hands, there is neither light-dependent nor GTP-dependent PLA2 activity in intact purified ROS. We also failed to detect PLA1 activity in those ROS preparations. Nevertheless, we detected significant amounts of PLA2 activity in two subretinal fractions adjacent to ROS: RPE (enriched with retinal pigment epithelial cells) and P200 (presumably containing neuronal cells, Muller cells, and rod inner segments). The enzyme present both in RPE and P200 is light- and GTP-independent, Ca2+- and Mg2+-independent, and seems to be optimally active in the alkaline pH range. Our results suggest that there is, if any, vanishingly little PLA2 or PLA1 activity in intact purified ROS and that the activity levels previously reported in the literature could have been due to a contamination by either RPE or P200. This is supported by our observation that some contaminated ROS preparations were "PLA2 active." PMID- 8702601 TI - Cloning and expression of sialidase L, a NeuAcalpha2-->3Gal-specific sialidase from the leech, Macrobdella decora. AB - Sialidase L is a NeuAcalpha2-->3Gal linkage-specific sialidase that releases 2,7 anhydro-NeuAc instead of NeuAc from sialoglycoconjugates (Chou, M.-Y., Li, S.-C., Kiso, M., Hasegawa, A., and Li, Y.-T.(1994) J. Biol. Chem. 269, 18821-18826). A 2. 5-kilobase cDNA of sialidase L was cloned by a combination of methods based on polymerase chain reactions. The composite cDNA sequence reveals an open reading frame coding for 762 amino acids, including a putative 28-residue signal peptide at the N terminus that is similar to the signal sequence of the Clostridium septicum sialidase. The result suggests that sialidase L is a secretory enzyme. The coding sequence excluding the putative signal peptide of sialidase L was overexpressed in Escherichia coli. The purified recombinant enzyme was characterized to be as active as the enzyme isolated from the leech. It also possessed the strict NeuAcalpha2-->3Gal linkage specificity and released the unique cleavage product, 2,7-anhydro-NeuAc from sialoglycoconjugates. The deduced amino acid sequence of sialidase L exhibits little similarity with other reported sialidases. However, sialidase L contains a conserved "FRIP region" and four repeating "Asp box" motifs that align well with the corresponding positions of bacterial sialidases. The predicted beta-strand structures near the conserved motifs of sialidase L are similar to those of Salmonella typhimurium sialidase. Several conserved single amino acid residues of bacterial sialidases, including those known to be involved in the active site of Salmonella enzyme, are conserved in the deduced amino acid sequence of sialidase L. This observation suggests that part of the catalytic mechanism of sialidase L may be similar to the ordinary sialidase. PMID- 8702602 TI - Identification of essential residues for the catalytic function of 85-kDa cytosolic phospholipase A2. Probing the role of histidine, aspartic acid, cysteine, and arginine. AB - Cytosolic phospholipase A2 (cPLA2) hydrolyzes the sn-2-acyl ester bond of phospholipids and shows a preference for arachidonic acid-containing substrates. We found previously that Ser-228 is essential for enzyme activity and is likely to function as a nucleophile in the catalytic center of the enzyme (Sharp, J. D., White, D. L., Chiou, X. G., Goodson, T., Gamboa, G. C., McClure, D., Burgett, S., Hoskins, J., Skatrud, P. L., Sportsman, J. R., Becker, G. W., Kang, L. H., Roberts, E. F., and Kramer, R. M.(1991) J. Biol. Chem. 266, 14850-14853). cPLA2 contains a catalytic aspartic acid motif common to the subtilisin family of serine proteases. Substitution within this motif of Ala for Asp-549 completely inactivated the enzyme, and substitutions with either glutamic acid or asparagine reduced activity 2000- and 300-fold, respectively. Additionally, using mutants with cysteine replaced by alanine, we found that Cys-331 is responsible for the enzyme's sensitivity to N-ethylmaleimide. Surprisingly, substituting alanine for any of the 19 histidines did not produce inactive enzyme, demonstrating that a classical serine-histidine-aspartate mechanism does not operate in this hydrolase. We found that substituting alanine or histidine for Arg-200 did produce inactive enzyme, while substituting lysine reduced activity 200-fold. Results obtained with the lysine mutant (R200K) and a coumarin ester substrate suggest no specific interaction between Arg-200 and the phosphoryl group of the phospholipid substrate. Arg-200, Ser-228, and Asp-549 are conserved in cPLA2 from six species and also in four nonmammalian phospholipase B enzymes. Our results, supported by circular dichroism, provide evidence that Asp-549 and Arg-200 are critical to the enzyme's function and suggest that the cPLA2 catalytic center is novel. PMID- 8702603 TI - Regulation of the kinetics of phosducin phosphorylation in retinal rods. AB - Phosducin (Pd) is a widely expressed phosphoprotein that regulates G-protein (G) signaling. Unphosphorylated Pd binds to Gbetagamma subunits and blocks their interaction with Galpha. This binding sequesters Gbetagamma and inhibits both receptor-mediated activation of Galpha and direct interactions between Gbetagamma and effector enzymes. When phosphorylated by cAMP-dependent protein kinase, Pd does not affect these functions of Gbetagamma. To further understand the role of Pd in regulating G-protein signaling in retinal rod photoreceptor cells, we have measured the abundance of Pd in rods and examined factors that control the rate of Pd phosphorylation. Pd is expressed at a copy number comparable to that for the rod G-protein, transducin (Gt). The ratio of rhodopsin (Rho) to Pd is 15. 5 +/- 3.5 to 1. The rate of Pd phosphorylation in rod outer segment preparations was dependent on [cAMP]. K1/2 for cAMP was 0.56 +/- 0. 09 microM, and the maximal rate of phosphorylation was approximately 500 pmol PO4 incorporated/min/nmol Rho. In the presence of Gtbetagamma this rate was decreased approximately 50-fold. From these data, one can estimate a t1/2 of approximately 3 min for the rephosphorylation of Pd in rods during the recovery period after a light response. This relatively slow rephosphorylation of the Pd.Gtbetagamma complex may provide a period of molecular memory in which sensitivity to further light stimuli is reduced as a result of sequestration of Gtbetagamma by Pd. PMID- 8702604 TI - Cannabinoid receptors are coupled to nitric oxide release in invertebrate immunocytes, microglia, and human monocytes. AB - The present study demonstrates that stereoselective binding sites for anandamide, a naturally occurring cannabinoid substance, can be found in invertebrate immunocytes and microglia. The anandamide-binding site is monophasic and of high affinity, exhibiting a Kd of 34.3 nM with a Bmax of 441 fmol/mg protein. These sites are highly selective, as demonstrated by the inability of other types of signaling molecules to displace [3H]anandamide. Furthermore, this binding site is coupled to nitric oxide release in the invertebrate tissues examined as well as in human monocytes. Interestingly, the cannabinoid-stimulated release of nitric oxide initiates cell rounding. Thus, these cannabinoid actions resemble those of opiate alkaloids. In this regard, we demonstrate that these signaling systems use the same effector system, i.e. nitric oxide release, but separate receptors. Last, the presence of a cannabinoid receptor in selected evolutionary diverse organisms indicates that this signaling system has been conserved for more than 500 million years. PMID- 8702605 TI - Secondary structure and tertiary fold of the birch pollen allergen Bet v 1 in solution. AB - Bet v 1 is the major birch pollen allergen and therefore the main cause of type I allergies observed in early spring. It is composed of 159 amino acid residues adding up to a molecular mass of 17 kDa. We determined the secondary structure and tertiary fold of full-length Bet v 1 by NMR spectroscopy. Two- and three dimensional NMR measurements suggest that Bet v 1 is a globular monomer in solution with a high content of well defined secondary structure. Of the total of 159 residues, 135 could be sequentially assigned, using an improved assignment strategy based mainly on heteronuclear experiments. An improved strategy for structure calculation revealed three helices and two beta-sheets as major elements of secondary structure. The globular tertiary structure is mainly stabilized by two antiparallel beta-sheets. The two helices at the C terminus are in accordance with the results from the solution structure of the chemically synthesized peptide Bet v 1-(125-154). This peptide is composed of two helices connected by a hinge. The structural features of Bet v 1 are highly similar to those found in the Ambrosia allergen Amb t V. PMID- 8702606 TI - The sphingomyelin-ceramide signaling pathway is involved in oxidized low density lipoprotein-induced cell proliferation. AB - Development of atherosclerosis is believed to involve proliferation of smooth muscle cells (SMC). Our laboratory previously demonstrated that the growth of bovine aortic SMC was stimulated by mildly oxidized low density lipoproteins (oxLDL) and that the mitogenic effect of oxLDL was greater than that induced by native LDL (Auge, N., Pieraggi, M. T., Thiers, J. C., Negre-Salvayre, A., and Salvayre R.(1995) Biochem. J. 309, 1015-1020). Since the lipid mediator ceramide has been described to be proliferative, the present work aimed at studying the potential involvement of the so-called sphingomyelin-ceramide pathway in the signal transduction cascade induced by oxLDL. Incubation of SMC with UV-oxidized LDL induced sphingomyelin hydrolysis (32%), which peaked at 60 min and was accompanied by a concomitant increase of intracellular ceramide level. The effect of oxidized LDL on sphingomyelin turnover exhibited the same LDL dose dependence as their mitogenic effect. Exogenous bacterial sphingomyelinase induced sphingomyelin hydrolysis and ceramide generation and also stimulated cell growth, in contrast to exogenous phospholipases A2, C, or D. This mitogenic effect was reproduced by incubating the cells with the cell-permeant ceramides, N-acetyl- and N-hexanoylsphingosines. Altogether, these data strongly suggest for the first time that activation of the sphingomyelin-ceramide pathway may play a pivotal role in the oxLDL-induced SMC proliferation and atherogenesis. PMID- 8702607 TI - A conformational rearrangement upon binding of IgE to its high affinity receptor. AB - One of the critical steps in the allergic reaction is the binding of immunoglobulin E (IgE) to its high affinity receptor (FcepsilonRI). FcepsilonRI is a tetrameric complex composed of an alpha-chain, a beta-chain, and a dimeric gamma-chain. The extracellular portion of the alpha-chain (alpha-t) is sufficient for the binding of IgE. The Fc portion of IgE contains two copies of the FcepsilonRI binding sites. In contrast, the binding stoichiometry is 1:1. Previously, it was hypothesized that the binding of FcepsilonRI to IgE results in a conformational change in IgE that precludes the binding of a second molecule (Presta, L., Shields, R., O'Connel, L., Lahr, S., Porter, J. , Gorman, C., and Jardieu, P.(1994) J. Biol. Chem. 269, 26368-26373). Here we characterize the secondary structure of IgE and alpha-t and analyze their interaction by circular dichroism spectroscopy. Binding experiments show that when IgE interacts with alpha-t there is a 15-26% decrease of the negative ellipticity at 217 nm. Together, the absence of an alpha-helix element in alpha-t and the small contribution of alpha-t to the spectra of the complex indicate that upon binding, a major conformational rearrangement must occur on IgE. In addition, we analyze the thermal unfolding of alpha-t, IgE, and their complex. Despite the several domains that constitute IgE and alpha-t, these molecules unfold cooperatively with two-state kinetics. PMID- 8702608 TI - Localization of mRNAs encoding Ca2+-inhibitable adenylyl cyclases along the renal tubule. Functional consequences for regulation of the cAMP content. AB - Expression of Ca2+-inhibitable types V and VI adenylyl cyclases was studied by reverse transcription-polymerase chain reaction in rat renal glomeruli and nephron segments isolated by microdissection. Quantitation of each mRNA was achieved using a mutant cRNA which differed from the wild type by substituting two bases to create a new restriction site in the corresponding cDNA. Type VI mRNA was present all along the nephron but was more abundant in distal than in proximal segments. The expression of type V mRNA was restricted to the glomerulus and to the initial portions of the collecting duct. Expression of the Ca2+ insensitive type IV mRNA studied on the same samples was evidenced only in the glomerulus. The functional relevance of the expression of Ca2+-inhibitable isoforms was studied by measuring cAMP content in the microdissected outer medullary collecting duct which expressed both type V mRNA (2367 +/- 178 molecules/mm tubular length; n = 8) and type VI mRNA (5658 +/- 543 molecules/mm, n = 8). Agents known to increase intracellular Ca2+ in this segment induced a Ca2+-dependent inhibition on either arginine vasopressin- or glucagon-stimulated cAMP level. The characteristics of these inhibitions suggest a functional and differential expression of types V and VI adenylyl cyclases in two different cell types of the rat outer medullary collecting duct. PMID- 8702609 TI - Assembled pre-B cell receptor complexes are retained in the endoplasmic reticulum by a mechanism that is not selective for the pseudo-light chain. AB - The pre-B cell receptor (BCR) complex, consisting of micro heavy chain, a pseudo light chain, and the Mb-1/B29 heterodimer, directs the transition to the mature B cell stage. Plasma membrane expression of the pre-BCR is extremely low, despite its presumed signaling function. We have compared assembly and intracellular transport of the pre-BCR complex with that of the BCR complex in mature B cells. Synthesis and assembly rate of pre-BCR and BCR components are comparable. However, the pre-BCR is subject to a highly efficient retention mechanism, which only allows exit of a few percent of the complexes from the endoplasmic reticulum (ER). This small transported pool of pre-BCR complexes is significantly enriched for protein-tyrosine kinase activity, as compared with the ER-localized receptor pool. Accordingly, the Src-related tyrosine kinase Lyn was found in the transported glycoprotein fraction but not in association with ER-localized glycoproteins. Upon introduction of a conventional light chain into pre-B cells, plasma membrane receptor levels increased, but the efficiency of intracellular transport of the receptor complex was not restored to that in mature B cells. This indicates that the ER retention mechanism is not selective for the pseudo light chain and may be inherent to pre-B cells. We propose that this retention mechanism contributes to the regulation of pre-BCR-mediated signal transduction. PMID- 8702610 TI - A role in enzyme activation for the N-terminal leader sequence in calmodulin. AB - We have found that deletion of residues 2-8 from the N-terminal leader sequence: Ala1-Asp2-Gln-Leu4-Thr-Glu6-Glu-Gln8, in calmodulin abolishes calmodulin dependent activation of skeletal muscle myosin light chain kinase activity and reduces calmodulin-dependent activation of smooth muscle myosin light chain kinase activity to approximately 50% of the maximum level measured at a saturating calmodulin concentration. Calmodulin-dependent activation of cerebellar nitric oxide synthase activity is not affected by this deletion. Overlapping tripeptide deletions from the leader sequence indicate that the acidic cluster, Glu6-Glu-Gln8, contains the determinants necessary for activation of myosin light chain kinase activity. Deletion of Asp2-Gln-Leu4 has no effect on activation of enzyme activity. Based on enzyme kinetic analyses, deletions in the leader sequence have little or no effect on the apparent affinities of calmodulin for the synthase or the two kinases. Since the N-terminal leader does not appear to play a significant structural role in the complexes between calmodulin and peptides representing the calmodulin-binding domains in the two kinases, our results indicate that it participates in secondary interactions with these enzymes that are important to activation, but not to recognition or binding of calmodulin. PMID- 8702611 TI - The luteinizing hormone/chorionic gonadotropin receptor has distinct transmembrane conductors for cAMP and inositol phosphate signals. AB - The luteinizing hormone/chorionic gonadotropin receptor is a member of the seven transmembrane receptor family. It is coupled, presumably via Gs and Gq, to two signal pathways involving adenylyl cyclase/cAMP and phospholipase C/inositol phosphate (IP). Little is known about the events prior to G-protein coupling: for example, whether these signals are generated from a single or multiple independent origins and mechanisms, when and where they diverge, and how they are transduced. We report novel observations that the cAMP signal and the IP signal originate and diverge upstream of G-protein coupling. The generation of these two signals independently involves Lys583 in exoloop 3 of the rat receptor. For this study, Lys583 of the receptor was substituted with a panel of amino acids, and mutant receptors were assayed for hormone binding and induction of cAMP, inositol monophosphate, inositol bisphosphate, and inositol trisphosphate. No substitutions for Lys583 were permissible for cAMP induction, despite successful surface expression and hormone binding. In contrast, several substitutions were permissible for IP induction. Our results suggest two distinct transmembrane signal conductors for cAMP and inositol phosphate signals and imply particular models of receptor activation not previously suggested. PMID- 8702612 TI - Identification of the alpha chain lysine donor sites involved in factor XIIIa fibrin cross-linking. AB - Biochemical studies of fibrin cross-linking were conducted to identify the specific Aalpha chain lysine residues that potentially serve as Factor XIIIa amine donor substrates during alpha polymer formation. A previously characterized Factor XIIIa fibrin lysine labeling system was employed to localize sites of donor activity based on their covalent incorporation of a synthetic peptide acceptor substrate analog modelled after the NH2-terminal cross-linking domain of alpha2 antiplasmin. Peptide-decorated fibrin was prepared using purified fibrinogen as the starting material. Cyanogen bromide digestion, immunoaffinity chromatography, high pressure liquid chromatography (HPLC), and enzyme-linked immunosorbent assay (anti-peptide) methodologies were employed to isolate purified CNBr fibrin fragments whose structures included the acceptor probe in cross-linked form and, therefore, represented regions of (amine) donor activity. Five alpha chain CNBr fragments (within Aalpha 208-610) and one gamma chain CNBr fragment (gamma 385-411) were the only portions of fibrin found associated with the acceptor peptide, based on collective sequencing, mass, and compositional data. Trypsin digestion, HPLC, and enzyme-linked immunosorbent assay (anti peptide) methodologies were used to isolate smaller derivatives whose structures included an alpha chain tryptic cleavage product (the donor arm) cross-linked to the trypsin-resistant synthetic peptide (the acceptor arm). Biochemical characterization and quantitative peptide recovery data revealed that 12 of the 23 potential lysine donor residues within alpha 208-610 had incorporated the peptide probe, whereas gamma chain donor activity was due solely to peptide cross linking at (gamma) Lys406; the alpha chain lysines, Lys556 and Lys580, accounted for 50% of the total alpha chain donor cross-linking activity observed, with Lys539, Lys508, Lys418, and Lys448 contributing an additional 28% and Lys601, Lys606, Lys427, Lys429, Lys208, Lys224, and/or Lys219 responsible for the remaining proportion (2-5%, each). The collective findings extend current models proposed for the mechanism of alpha polymer formation, raise questions concerning the physiological role of multiple alpha chain donor sites, and, most importantly, provide specific information that should facilitate future efforts to identify the respective lysine and glutamine partners involved in native fibrin alpha chain cross-linking. PMID- 8702613 TI - Mode of action of the antimicrobial peptide indolicidin. AB - Indolicidin is a cationic antimicrobial peptide isolated from bovine neutrophils. It consists of only 13 amino acids, has the highest tryptophan content of any known protein, and is amidated at the carboxyl terminus in nature. By circular dichroism spectroscopy a weak poly-L-proline II extended helix structure was observed that became substantially more pronounced upon interaction with liposomes. Indolicidin bound purified surface lipopolysaccharide with high affinity and permeabilized the outer membrane of Escherichia coli to the small hydrophobic molecule 1-N-phenylnapthylamine (Mr 200), results consistent with indolicidin crossing the outer membrane via the self-promoted uptake pathway. The methyl esterification of indolicidin's carboxyl terminus increased its activity for Gram-negative and Gram-positive bacteria. In Gram-negative bacteria this was associated with an increased binding to lipopolysaccharide and increased permeabilization of the outer membrane. The cytoplasmic membrane was the site of action of indolicidin as assayed in E. coli by the unmasking of cytoplasmic beta galactosidase due to membrane permeabilization. The mechanism for this activity was shown to be the ability of the peptide to cause an increase in the transmembrane current of planar lipid bilayers. This current increase was activated by transmembrane potentials in excess of -70 to -80 mV. Consistent with this, there was a substantial decrease in indolicidin-mediated bacterial killing and permeabilization of the cytoplasmic membrane of E. coli that had been pretreated with the uncoupler carbonyl cyanide-m-chlorophenyl hydrazone. In planar bilayers, indolicidin induced the formation of discrete channels, which ranged in conductance from 0.05-0.15 nS. Thus despite the small size and unique composition of indolicidin, it was capable of killing Gram-negative bacteria by crossing the outer membrane and causing disruption of the cytoplasmic membrane by channel formation. PMID- 8702614 TI - The IGF-I receptor gene promoter is a molecular target for the Ewing's sarcoma Wilms' tumor 1 fusion protein. AB - Desmoplastic small round cell tumor (DSRCT) is an abdominal malignancy in children which is characterized by a recurrent chromosomal translocation, t(11;22)(p13;q12). This rearrangement results in the fusion of the ubiquitously expressed EWS1 gene to the Wilms' tumor suppressor (WT1) gene. The chimeric protein contains the N-terminal domain of EWS1 fused to the DNA-binding domain of WT1, including zinc fingers 2-4. Because WT1 has been shown previously to bind and repress the insulin-like growth factor I (IGF-I-R) promoter, we investigated whether this promoter is, in addition, a target for the aberrant EWS/WT1 transcription factor. EWS/WT1 activated the IGF-I-R promoter approximately 340%, whereas a fusion protein containing a three-amino acid insert (KTS) between zinc fingers 3 and 4 had no effect. On the other hand, expression vectors encoding either WT1 or EWS1 reduced the activity of the promoter to 46 and 58% of control values, respectively. Results of gel shift assays indicate that the binding affinity of EWS/WT1 to a fragment of the 5'-flanking region of the receptor promoter was higher than the affinity of WT1 itself. Consistent with the results of functional assays, the binding of EWS/WT1(+KTS) was significantly reduced. Due to the central role of the IGF-I-R in tumorigenesis, activation of the receptor promoter by EWS/WT1 may constitute a potential mechanism for the etiology and/or progression of DSRCT. PMID- 8702615 TI - Genomic organization of the mouse and human genes for vascular endothelial growth factor B (VEGF-B) and characterization of a second splice isoform. AB - A second isoform and the genomic structures of mouse and human vascular endothelial growth factor B are described. Both genes consist of seven coding exons and span about 4 kilobases of DNA. The two identified isoforms of vascular endothelial growth factor B are generated by alternative splicing where different splice acceptor sites in exon 6 introduce a frameshift and a partial use of different but overlapping reading frames. Consequently, the COOH-terminal domains in the two isoforms show no resemblance. Mouse and human cDNA clones for the novel isoform of vascular endothelial growth factor B encoded a secreted protein of 186 amino acid residues. Expression in transfected cells generated a protein of 25 kDa which upon secretion was modified by O-linked glycosylation and displayed a molecular mass of 32 kDa under reducing conditions. The protein was expressed as a disulfide-linked homodimer, and heterodimers were generated when coexpressed with vascular endothelial growth factor. The entirely different COOH terminal domains in the two isoforms of vascular endothelial growth factor B imply that some functional properties of the two proteins are distinct. PMID- 8702616 TI - Cell cycle arrest promotes trans-hammerhead ribozyme action in yeast. AB - A hammerhead ribozyme designed to cleave the yeast ADE1 mRNA has been expressed in yeast under the control of a galactose-inducible promoter. RNA prepared from the galactose-induced yeast cultures possesses an activity that cleaves ADE1 mRNA in vitro. However, in spite of high expression levels of the ribozyme, no cleavage activity could be demonstrated in vivo. On the other hand, when the yeast cells expressing hammerhead RNA were treated with the alpha-factor mating pheromone, the level of ADE1 mRNA was reduced by 50%. Similar reductions were observed when this strain was cultured in the presence of lithium acetate or in nitrogen-free medium. Moreover, control experiments in which disabled hammerhead genes were expressed showed no such reductions. Extension of the length of the flanking recognition arms of the ribozyme from a total of 10 to 16 or 24 nucleotides diminished the inhibitory effect of the ribozyme. These data suggest that ribozymes are able to cleave a trans-RNA target in yeast. PMID- 8702617 TI - Cytoplasmic domain of natriuretic peptide receptor-C inhibits adenylyl cyclase. Involvement of a pertussis toxin-sensitive G protein. AB - Natriuretic peptide receptor C (NPR-C) is a disulfide-linked homodimer with an approximately 440-amino acid extracellular domain and a 37-amino acid cytoplasmic domain; it functions in the internalization and degradation of bound ligand. The use of NPR-C-specific natriuretic peptide analogs has implicated this receptor in mediating the inhibition of adenylyl cyclase or activation of phospholipase C. In the present studies we have investigated the role of the cytoplasmic domain of NPR-C in signaling the inhibition of adenylyl cyclase. Polyclonal rabbit antisera were raised against a 37-amino acid synthetic peptide (R37A) corresponding to the cytoplasmic domain of NPR-C. Incubation of anti-R37A with rat heart particulate fractions blocked atrial natriuretic peptide-dependent inhibition of adenylyl cyclase. The cytoplasmic domain peptides R37A and TMC (10 residues of transmembrane domain appended on R37A) were equipotent in inhibiting adenylyl cyclase (Ki approximately 1 nM) in a GTP-dependent manner, whereas K37E (a scrambled peptide control for R37A) did not inhibit adenylyl cyclase activity. Prior incubation of membranes with pertussis toxin blocked R37A or TMC inhibition of cAMP production. Detergent solubilization of the rat heart particulate fraction destroyed natriuretic peptide inhibition of adenylyl cyclase, but TMC was able to inhibit cAMP production in a dose-dependent manner. Our results provide evidence that the 37-amino acid cytoplasmic domain of NPR-C is sufficient for signaling inhibition of adenylyl cyclase through a pertussis toxin-sensitive G protein. PMID- 8702618 TI - Mutational analysis of RNA structures and sequences postulated to affect 3' processing of M1 RNA, the RNA component of Escherichia coli RNase P. AB - When the rnpB gene encoding M1 RNA, the RNA component of Escherichia coli RNase P, is transcribed, the primary M1 RNA transcript (pM1 RNA) is produced and subsequently processed at the 3' end to generate the mature M1 RNA. To study features of pM1 RNA thought to be involved in RNA processing, systematic mutations were introduced in sequence elements and secondary structures surrounding the processing site using p23 RNA, a truncated pM1 RNA transcribed from the internally deleted rnpB gene, as a model substrate and the processing of its mutant derivatives was analyzed in vivo and in vitro. Neither the alteration of two bases forming the processing site nor the disruption of secondary structures surrounding the site significantly affected the processing efficiency although the secondary structures were required for maintaining RNA stability. In contrast, mutations at the rne-dependent site, GAUUU, immediately 3' to the processing site inhibited the processing and the extent of the inhibition varied with the altered sequences. Furthermore, the processing of the mutants of the rne dependent site as well as wild-type p23 RNA was inhibited in an E. coli rnets strain at the nonpermissive temperature. PMID- 8702619 TI - Protein cross-linking by the Maillard reaction. Isolation, characterization, and in vivo detection of a lysine-lysine cross-link derived from methylglyoxal. AB - The Maillard reaction, initiated by nonenzymatic glycosylation of amino groups on proteins by reducing sugars, has been studied for its potential role in aging and the complications of diabetes. One of the major consequences of the advanced Maillard reaction in proteins is the formation of covalently cross-linked aggregates. The chemical nature of the cross-linking structures is largely unknown. Recently, methylglyoxal has been shown to be a potential glycating agent in vivo and suggested to be a common intermediate in the Maillard reaction involving glucose. Methylglyoxal can form enzymatically or nonenzymatically from glycolytic intermediates and by retro-aldol cleavage of sugars. Its elevation in tissues in diabetes and its high potency to glycate and cross-link proteins led us to investigate the chemical nature of its advanced Maillard products. Using an approach in which a synthetic model peptide was reacted with methylglyoxal, we isolated and purified a cross-linked peptide dimer. Characterization of this dimer revealed that the peptides are linked through epsilon amino groups of lysine residues. The actual cross-link was shown to be a methylimidazolium, formed from the reaction of two lysines and two methylglyoxal molecules. We have named this cross-link imidazolysine. Imidazolysine was detected in proteins by high performance liquid chromatography using a postcolumn derivatization method. Proteins incubated with methylglyoxal showed a time-dependent formation of imidazolysine. Quantification of imidazolysine in human serum proteins revealed a significant increase (p < 0.05) in diabetic samples (mean +/- S.D., 313.8 +/- 52.7 pmol/mg protein) when compared with normal samples (261.3 +/- 50.4). These values correlated with glycohemoglobin (p < 0.05). These results provide chemical evidence for protein cross-linking by dicarbonyl compounds in vivo. PMID- 8702620 TI - Inactivation of EF-hands makes GCAP-2 (p24) a constitutive activator of photoreceptor guanylyl cyclase by preventing a Ca2+-induced "activator-to inhibitor" transition. AB - Guanylyl cyclase activator proteins GCAP-1 and GCAP-2 (Dizhoor et al. , 1995, Gorczyca et al., 1995) are members of a recently identified subclass of EF-hand type Ca2+-binding proteins that respond to Ca2+ differently than any other known members of the EF-hand superfamily. GCAPs acquire an activating conformation only in their Ca2+-free form. Free Ca2+ concentrations corresponding to levels in dark adapted vertebrate photoreceptors inhibit the ability of GCAPs to activate photoreceptor guanylyl cyclases (RetGCs). We studied the effects of mutations that block binding of Ca2+ to the EF-hands of GCAP-2. Unlike other EF-hand proteins, which fail to activate their target when their EF-hands are inactivated by mutations, GCAP-2 with any single EF-hand inactivated remains active and is 3 6 times less sensitive to the inhibitory effect of Ca2+. Inactivation of any two or all three EF-hands produces active forms of GCAP-2 that are insensitive to inhibition by physiological intracellular concentrations of Ca2+. Unexpectedly we also found that activation of RetGCs by a Ca2+-insensitive mutant is inhibited by Ca2+-loaded wild type GCAP-2. We propose the following. 1) GCAP-2 can exist in two extreme functional forms: an apo form that activates RetGCs and a Ca2+-loaded form that blocks activation of RetGCs. 2) All three EF-hands of GCAP-2 contribute to the inhibitory effect of Ca2+. 3) Inactivation of two or three EF-hands is sufficient to shift the "activator-inhibitor" transition outside the physiological range of intracellular free Ca2+. PMID- 8702621 TI - Hormonal and environmental regulation of a plant PDR5-like ABC transporter. AB - The PDR5 gene from yeast encodes an ABC (ATP-binding cassette) transporter involved in the ATP-dependent efflux of a variety of structurally unrelated cytotoxic compounds. We report here on the cDNA cloning and characterization of a PDR5 homolog (TUR2) from a higher eukaryote, the aquatic plant Spirodela polyrrhiza. We show that TUR2 transcripts accumulate throughout the plant following treatment with the steroid-like hormone, abscisic acid, and that this induction can be repressed by the adenine-derived hormone, kinetin. Furthermore, TUR2 gene expression is induced by environmental stress treatments such as low temperature and high salt. These data indicate that PDR5 homologs are present in plants, that they may function during stress conditions in an analogous fashion to that described in yeast, and that the expression of such ABC transporters is subject to a complex hormonal and environmental regulation. PMID- 8702622 TI - Brain cytoplasmic and flagellar outer arm dyneins share a highly conserved Mr 8,000 light chain. AB - Sequence comparisons with the Mr 8,000 light chain from Chlamydomonas outer arm dynein revealed the presence of highly conserved homologues (up to 90% identity) in the expressed sequence tag data base (King, S. M. & Patel-King, R. S. (1995a) J. Biol. Chem. 270, 11445-11452). Several of these homologous sequences were derived from organisms and/or tissues that lack motile cilia/flagella, suggesting that these proteins may function in the cytoplasm. In Drosophila, lack of the homologous protein results in embryonic lethality (Dick, T., Ray, K., Salz, H. K. & Chia, W.(1996) Mol. Cell. Biol., 16, 1966-1977). Fractionation of mammalian brain homogenates reveals three distinct cytosolic pools of the homologous protein, one of which specifically copurifies with cytoplasmic dynein following both ATP-sensitive microtubule affinity/sucrose density gradient centrifugation and immunoprecipitation with a monoclonal antibody specific for the 74-kDa intermediate chain (IC74). Quantitative densitometry indicates that there is one copy of the Mr 8,000 polypeptide per IC74. Dual channel confocal immunofluorescent microscopy revealed that the Mr 8,000 protein is significantly colocalized with cytoplasmic dynein but not with kinesin in punctate structures (many of which are associated with microtubules) within mammalian oligodendrocytes. Thus, it appears that flagellar outer arm and brain cytoplasmic dyneins share a highly conserved light chain polypeptide that, at least in Drosophila, is essential for viability. PMID- 8702623 TI - Fusion of insulin receptor ectodomains to immunoglobulin constant domains reproduces high-affinity insulin binding in vitro. AB - A unique feature of the insulin receptor is that it is dimeric in the absence of ligand. Dimerization of two adjacent transmembrane domain (TMD) alpha helices has been shown to be critical in receptor kinase activation. Moreover, previous work has suggested that the TMD is involved in stabilizing the high-affinity binding site; soluble receptors expressed after simple truncation at the ectodomain-TMD junction have reduced affinity for insulin. To further examine this issue, we have replaced the TMD and intracellular domain of the soluble human insulin receptor (HIRs) with constant domains from immunoglobulin Fc and lambda subunits (HIRs-Fc and HIRs-lambda). Studies of receptor biosynthesis and binding characteristics were performed following transient transfection of receptor cDNAs into human embryonal kidney 293 cells. Each hybrid receptor was initially synthesized as a single chain proreceptor, followed by cleavage into alpha- and beta-Fc or beta-lambda subunits. The majority of secreted protein appeared in the cell medium as fully processed heterotetramer. Fc fragments released from HIRs-Fc by papain digestion and analyzed by nonreducing SDS-polyacrylamide gel electrophoresis were dimeric. Furthermore, dissociation constants for both chimeras were similar to those for the full-length holoreceptor (wild-type receptor, Kd1 = 200 pM and Kd2 = 2 nM; HIRs-Fc, Kd1 = 200 pM and Kd2 = 40 nM; and HIRs-lambda, Kd1 = 200 pM and Kd2 = 5 nM). These results extend previous observations that dimerization of the membrane-proximal ectodomain is necessary to maintain an intact high-affinity insulin-binding site. PMID- 8702624 TI - Comparison of the rat nucleolar protein nopp140 with its yeast homolog SRP40. Differential phosphorylation in vertebrates and yeast. AB - Rat Nopp140, a nonribosomal protein of the nucleolus and coiled bodies, was characterized as one of the most highly phosphorylated proteins in the cell. Based on its subcellular location, its nuclear localization signal binding capacity, and its shuttling between the nucleolus and the cytoplasm, Nopp140 was proposed to function as a chaperone in ribosome biogenesis. This study shows that casein kinase II phosphorylates Nopp140 to its unusual high degree and identifies the yeast SRP40 gene product as immunologically and structurally related to rat Nopp140. SRP40 encodes an acidic (pI = 3. 9), serine-rich (49%) protein of 41 kDa whose carboxyl terminus exhibits 59% sequence identity to that of Nopp140. SRP40 localizes to the yeast nucleolus and is required at a specific cellular concentration for optimal growth as indicated by the negative effect on cell growth of both overexpression and deletion of its gene. Like Nopp140, SRP40 is phosphorylated by casein kinase II, but to a much lesser extent. While the parallels between these two proteins suggest that SRP40 is the bona fide yeast Nopp140 homolog, their disparities reflect the differences in nucleolar dynamics and regulation of ribosome biogenesis between yeast and vertebrates. PMID- 8702625 TI - Huntingtin is ubiquitinated and interacts with a specific ubiquitin-conjugating enzyme. AB - Using the yeast two-hybrid system, we have identified a human ubiquitin conjugating enzyme (hE2-25K) as a protein that interacts with the gene product for Huntington disease (HD) (Huntingtin). This protein has complete amino acid identity with the bovine E2-25K protein and has striking similarity to the UBC-1, -4 and -5 enzymes of Saccharomyces cerevisiae. This protein is highly expressed in brain and a slightly larger protein recognized by an anti-E2-25K polyclonal antibody is selectively expressed in brain regions affected in HD. The huntingtin E2-25K interaction is not obviously modulated by CAG length. We also demonstrate that huntingtin is ubiquitinated. These findings have implications for the regulated catabolism of the gene product for HD. PMID- 8702626 TI - Effects of deletion of the carboxyl-terminal domain of ApoA-I or of its substitution with helices of ApoA-II on in vitro and in vivo lipoprotein association. AB - In the present study, the lipoprotein association of apoA-I, an apoA-I (DeltaAla190-Gln243) deletion mutant and an apoA-I (Asp1-Leu189)/apoA-II (Ser12 Gln77) chimera were compared. At equilibrium, 80% of the 125I-labeled apolipoproteins associated with lipoproteins in rabbit or human plasma but with very different distribution profiles. High density lipoprotein (HDL)2,3 associated fractions were 0.60 for apoA-I, 0.30 for the chimera, and 0.15 for the deletion mutant, and corresponding very high density lipoprotein-associated fractions were 0.20, 0.50, and 0.65. Clearance curves after intravenous bolus injection of 125I-labeled apolipoproteins (3 microg/kg) in normolipemic rabbits could be adequately fitted with a sum of three exponential terms, yielding overall plasma clearance rates of 0.028 +/- 0.0012 ml.min-1 for apoA-I (mean +/- S.E.; n = 6), 0.10 +/- 0.008 ml.min-1 for the chimera (p < 0.001 versus apoA-I) and 0.38 +/- 0.022 ml.min-1 for the deletion mutant (p < 0.001 versus apoA-I and versus the chimera). Fractions that were initially cleared with a t1/2 of 3 min, most probably representing free apolipoproteins, were 0.30 +/- 0.04, 0.50 +/- 0.06 (p = 0.02 versus apoA-I), and 0.64 +/- 0.07 (p = 0.002 versus apoA-I), respectively. At 20 min after the bolus, the fractions of injected material associated with HDL2,3 were 0.55 +/- 0.06, 0.25 +/- 0.03 (p = 0.001 versus apoA I), and 0.09 +/- 0.01 (p < 0.001 versus apoA-I and versus the chimera), respectively, whereas the fractions associated with very high density lipoprotein were 0. 15 +/- 0.006, 0.25 +/- 0.03 (p = 0.008 versus apoA-I), and 0.27 +/- 0.03 (p = 0.003 versus apoA-I), respectively. The ability of the different apolipoproteins to bind to HDL3 particles and displace apoA-I in vitro were compared. The molar ratios at which 50% of 125I-labeled apoA-I was displaced from the surface of HDL3 particles were 1:1 for apoA-I, 3:1 for the chimera and 12:1 for the deletion mutant, indicating 3- and 12-fold reductions of the affinities for HDL3 of the chimera and the deletion mutant, respectively. These data suggest that the carboxyl-terminal pair of helices of apoA-I are involved in the initial rapid binding of apoA-I to the lipid surface of HDL. Although the lipid affinity of apoA-II is higher than that of apoA-I, substitution of the carboxyl-terminal helices of apoA-I with those of apoA-II only partially restores its lipoprotein association. Thus, this substitution may affect cooperative interactions with the middle amphipathic helices of apoA-I that are critical for its specific distribution over the different HDL species. PMID- 8702627 TI - Prk, a cytokine-inducible human protein serine/threonine kinase whose expression appears to be down-regulated in lung carcinomas. AB - We have cloned and characterized a putative protein serine/threonine kinase termed prk through a combination of polymerase chain reaction and conventional cDNA library screening approaches. There are apparently two distinct domains within prk protein deduced from its nucleotide sequences. The amino-terminal portion has the feature of the catalytic domain of a serine/threonine kinase and shows strong homology to mouse fnk and other polo family kinases including mouse snk, human and murine plk, Drosophila polo, and yeast Cdc5. The carboxyl-terminal portion, presumably the regulatory domain, shares extensive homology to mouse fnk. Northern blotting analyses reveal that prk expression is restricted to a very limited number of tissues with placenta, ovaries, and lung containing detectable amounts of prk mRNA. prk mRNA expression is also detected at a low level in the megakaryocytic cell line Dami, MO7e, and three brain glioma cell lines. In addition, refeeding of serum-deprived MO7e, Dami, and K562 cells of hematopoietic origin and GMOO637D of lung fibroblasts rapidly activates prk mRNA expression with its peak induction around 2 h after serum addition. prk gene activation by the serum requires no new protein synthesis. The recombinant cytokines such as interleukin-3 and thrombopoietin also activate prk mRNA expression in MO7e cells. Furthermore, a survey of RNAs isolated from the tumor and the uninvolved tissues from 18 lung cancer patients reveals that prk mRNA expression is significantly down-regulated in tumor tissues. Southern blotting analysis indicates that the prk gene is present in a single copy in the genome of tumors and normal cells. Taken together, these results suggest that prk expression may be restricted to proliferating cells and involved in the regulation of cell cycle progression. The molecular cloning of prk cDNA will facilitate the study of its biological role as well as its potential role in tumorigenesis. PMID- 8702628 TI - Covalent linkage between proteins of the inter-alpha-inhibitor family and hyaluronic acid is mediated by a factor produced by granulosa cells. AB - The direct interaction of hyaluronic acid (HA) and proteins of the inter-alpha inhibitor family plays a critical role in organization and stabilization of the expanding cumulus extracellular matrix (cECM) following an ovulatory stimulus. Despite similarities in the morphology of cumulus oocyte complexes (COCs) expanding in vivo and in vitro, we find that the cECM of COCs which expand within intact follicles are more elastic and resistant to shear stress than the cECM of those stabilized in vitro. Western blot analysis shows that only the heavy chains of inter-alpha-inhibitor are incorporated into the cECM and appears to be covalently linked to HA after stabilization in vivo while intact inter-alpha inhibitor is bound to the HA-enriched cECM by a non-covalent mechanism in in vitro stabilized COCs. However, purified pre-alpha-inhibitor and HA can form covalent linkage in the presence of granulosa cells or with granulosa cell conditioned medium. In addition, COCs resistance to shear stress is also enhanced by coincubation with granulosa cells. Upon formation of the apparent covalent linkage between heavy chains and HA in culture medium, the light chain (bikunin) is concomitantly released into the medium as a complex with chondroitin sulfate moieties of inter-alpha-inhibitor supporting the possibility that HA may replace the chondroitin sulfate linkage to the heavy chains. We speculate that a factor(s) secreted by granulosa cells within the follicle may catalyze a transesterification reaction resulting in an exchange of chondroitin sulfate with HA at the heavy chain/chondroitin sulfate junction followed by release of chondroitin sulfate-bikunin into the follicular fluid. It is also possible that the consequent further stabilization of the cECM through the covalent interaction of HA and heavy chains of inter-alpha-inhibitor may play an important role in the process of ovulation. PMID- 8702629 TI - Isolation and sequence of a novel human chondrocyte protein related to mammalian members of the chitinase protein family. AB - We describe the isolation of a novel protein from the conditioned medium of human articular cartilage chondrocytes in primary culture. This 39-kDa protein has the N-terminal sequence YKL, which we have termed YKL-39. The 1434-nucleotide sequence of the YKL-39 cDNA predicts a 385-residue initial translation product and a 364-residue mature YKL-39. The amino acid sequence of YKL-39 is most closely related to YKL-40, followed by macrophage chitotriosidase, oviductal glycoprotein, and macrophage YM-1. All five proteins share significant sequence identity with bacterial chitinases and have the probable structure of an (alphabeta)8 barrel. YKL-39 lacks the active site glutamate, which is essential for the activity of chitinases, and as expected has no chitinase activity. The highest level of YKL-39 mRNA expression is seen in chondrocytes, followed by synoviocytes, lung, and heart. YKL-39 accounts for 4% of the protein in chondrocyte-conditioned medium, prostromelysin accounts for 17%, and YKL-40 accounts for 33%. In contrast to YKL-40, YKL-39 is not a glycoprotein and does not bind to heparin. PMID- 8702630 TI - Conformational changes and stabilization of inosine 5'-monophosphate dehydrogenase associated with ligand binding and inhibition by mycophenolic acid. AB - The effects of substrate, product, and inhibitor (mycophenolic acid) binding on the conformation and stability of hamster type II inosine 5'-monophosphate dehydrogenase (IMPDH) have been examined. The protein in various states of ligand occupancy was compared by analyzing susceptibility to in vitro proteolysis, the degree of binding of a hydrophobic fluorescent dye, secondary structure content as determined by far-UV circular dichroism spectra, and urea-induced denaturation curves. These analysis methods revealed consistent evidence that IMPDH undergoes a local reorganization when IMP or XMP bind. NAD+ produced no such effect. In fact, no evidence was found for NAD+ binding independently of IMP. It is proposed that IMPDH adopts an open conformation around its nucleotide binding sites in the absence of substrates and that binding of IMP stabilizes a closed conformation that has a higher affinity for NAD+. The data also suggest the enzyme remains in the closed configuration throughout the catalytic steps and then reverts to the open conformation with XMP release, thereby consummating the enzyme cycle. Mycophenolic acid inhibition appeared to impart even greater stability. We propose that localized conformational changes occur during the normal and mycophenolic acid-inhibited reaction sequences of IMPDH. PMID- 8702631 TI - Inhibition of the 3' --> 5' exonuclease of human DNA polymerase epsilon by fludarabine-terminated DNA. AB - Incorporation of the anticancer drug fludarabine (9-beta-D-arabinofuranosyl-2 fluoroadenine 5'-monophosphate; F-ara-AMP) into the 3'-end of DNA during replication causes termination of DNA strand elongation and is strongly correlated with loss of clonogenicity. Because the proofreading mechanisms that remove 3'-F-ara-AMP from DNA represent a possible means of resistance to the drug, the present study investigated the excision of incorporated F-ara-AMP from DNA by the 3' --> 5'-exonuclease activity of DNA polymerase epsilon from human leukemia CEM cells. Using the drug-containing and normal deoxynucleotide oligomers (21-base) annealed to M13mp18(+) DNA as the excision substrates, we demonstrated that DNA polymerase epsilon was unable to effectively remove F-ara AMP from the 3'-end of the oligomer. However, 3'-terminal dAMP and subsequently other deoxynucleotides were readily excised from DNA in a distributive fashion. Kinetic evaluation demonstrated that although DNA polymerase epsilon has a higher affinity for F-ara-AMP-terminated DNA (Km = 7.1 pM) than for dAMP-terminated DNA of otherwise identical sequence (Km = 265 pM), excision of F-ara-AMP proceeded at a substantially slower rate (Vmax = 0.053 pmol/min/mg) than for 3'-terminal dAMP (Vmax = 1.96 pmol/min/mg). When the 3'-5' phosphodiester bond between F-ara-AMP at the 3'-terminus and the adjacent normal deoxynucleotide was cleaved by DNA polymerase epsilon, the reaction products appeared to remain associated with the enzyme but without the formation of a covalent bond. No further excision of the remaining oligomers was observed after the addition of fresh DNA polymerase epsilon to the reaction. Furthermore, the addition of DNA polymerase alpha and deoxynucleoside triphosphates to the excision reaction failed to extend the oligomers. After DNA polymerase epsilon had been incubated with 3'-F-ara-AMP-21 mer for 10 min, the enzyme was no longer able to excise 3'-terminal dAMP from a freshly added normal 21-mer annealed to M13mp18(+) template. We conclude that the 3' --> 5' exonuclease of human DNA polymerase epsilon can remove 3'-terminal F ara-AMP from DNA with difficulty and that this excision results in a mechanism mediated formation of "dead end complex." PMID- 8702632 TI - Covalent DNA binding by vaccinia topoisomerase results in unpairing of the thymine base 5' of the scissile bond. AB - We have used potassium permanganate to probe contacts between vaccinia DNA topoisomerase and thymine residues in its 5'-CCCTT downward arrow DNA target site. Two major conclusions emerge from the experiments presented: (i) permanganate oxidation of the +2T base of the scissile strand interferes with topoisomerase binding to DNA, and (ii) the +1T base of the scissile strand becomes unpaired upon formation of the covalent topoisomerase-DNA intermediate. Disruption of T:A base pairing is confined to the +1-position. Covalently bound DNAs that have experienced this structural distortion (such DNAs being marked by oxidation at +1T) are fully capable of being religated. We suggest that a protein induced DNA conformational change is a component of the strand passage step of the topoisomerase reaction. PMID- 8702633 TI - Role of c-Src tyrosine kinase in G protein-coupled receptor- and Gbetagamma subunit-mediated activation of mitogen-activated protein kinases. AB - Several G protein-coupled receptors that interact with pertussis toxin-sensitive heterotrimeric G proteins mediate Ras-dependent activation of mitogen-activated protein (MAP) kinases. The mechanism involves Gbetagamma subunit-mediated increases in tyrosine phosphorylation of the Shc adapter protein, Shc*Grb2 complex formation, and recruitment of Ras guanine nucleotide exchange factor activity. We have investigated the role of the ubiquitous nonreceptor tyrosine kinase c-Src in activation of the MAP kinase pathway via endogenous G protein coupled lysophosphatidic acid (LPA) receptors or by transient expression of Gbetagamma subunits in COS-7 cells. In vitro kinase assays of Shc immunoprecipitates following LPA stimulation demonstrated rapid, transient recruitment of tyrosine kinase activity into Shc immune complexes. Recruitment of tyrosine kinase activity was pertussis toxin-sensitive and mimicked by cellular expression of Gbetagamma subunits. Immunoblots for coprecipitated proteins in Shc immunoprecipitates revealed a transient association of Shc and c-Src following LPA stimulation, which coincided with increases in Shc-associated tyrosine kinase activity and Shc tyrosine phosphorylation. LPA stimulation or expression of Gbetagamma subunits resulted in c-Src activation, as assessed by increased c-Src autophosphorylation. Overexpression of wild-type or constitutively active mutant c-Src, but not kinase inactive mutant c-Src, lead to increased tyrosine kinase activity in Shc immunoprecipitates, increased Shc tyrosine phosphorylation, and Shc.Grb2 complex formation. MAP kinase activation resulting from LPA receptor stimulation, expression of Gbetagamma subunits, or expression of c-Src was sensitive to dominant negatives of mSos, Ras, and Raf. Coexpression of Csk, which inactivates Src family kinases by phosphorylating the regulatory C-terminal tyrosine residue, inhibited LPA stimulation of Shc tyrosine phosphorylation, Shc.Grb2 complex formation, and MAP kinase activation. These data suggest that Gbetagamma subunit-mediated formation of Shc.c-Src complexes and c-Src kinase activation are early events in Ras-dependent activation of MAP kinase via pertussis toxin-sensitive G protein-coupled receptors. PMID- 8702634 TI - Overproduction, purification, and characterization of the XPC subunit of the human DNA repair excision nuclease. AB - Xeroderma pigmentosum complementation group C gene (XPC) encodes a protein of 125 kDa which is present in a tight complex with a 58-kDa protein encoded by the human homolog of the yeast RAD23 gene, HHR23B (Masutani, C., Sugasawa, K., Yanagisawa, J., Sonoyama, T., Ui, M., Enomoto, T., Takio, K., Tanaka, K., van der Spek, P. J., Bootsma, D., Hoeijmakers, J. H. J., and Hanaoka, F.(1994) EMBO J. 13, 1831-1843). The XPC-HHR23B complex is required for excision of thymine dimers from DNA in a human excision nuclease system reconstituted from purified proteins. In order to understand the role of the XPC-HHR23B complex in excision repair, we have overexpressed each subunit alone and the heterodimer in heterologous systems, purified them, and characterized their biochemical properties. We find that both XPC and the heterodimer bind DNA with high affinity and UV-damaged DNA with slightly higher preference. Surprisingly, we find that the XPC subunit alone is sufficient for reconstitution of the human excision nuclease and that the HHR23B subunit has no detectable effect on the excision activity of the reconstituted system. PMID- 8702635 TI - Characterization of mouse ST8Sia II (STX) as a neural cell adhesion molecule specific polysialic acid synthase. Requirement of core alpha1,6-linked fucose and a polypeptide chain for polysialylation. AB - We previously showed that mouse ST8Sia II (STX) exhibits polysialic acid (PSA) synthase activity in vivo as well as in vitro (Kojima, N., Yoshida, Y., and Tsuji, S. (1995) FEBS Lett. 373, 119-122, 1995). In this paper, we reported that the neural cell adhesion molecule (NCAM) was specifically polysialylated by a single enzyme, ST8Sia II. PSA-expressing Neuro2a cells (N2a-STX) were established by stable transfection of the mouse ST8Sia II gene. Only the 140- and 180-kDa isoforms of NCAM in N2a-STX cells were specifically polysialylated in vivo, although other membrane proteins of N2a-STX were polysialylated in vitro. A recombinant soluble mouse ST8Sia II synthesized PSA on a recombinant soluble NCAM fused with the Fc region of human IgG1 (NCAM-Fc) as well as fetuin. However, NCAM Fc served as a 1500-fold better acceptor for ST8Sia II than fetuin. Treatment of NCAM-Fc with Charonia lampas alpha-fucosidase, which is able to cleave alpha1,6 linked fucose, clearly reduced the polysialylation of NCAM-Fc by ST8Sia II. PSA was not synthesized on the N-glycanase-treated NCAM-Fc polypeptide or the free N glycans of NCAM-Fc. When fetuin and its glycopeptide and N-glycans of fetuin were used as substrates for ST8Sia II, PSA was found to be synthesized on native fetuin and its glycopeptide but not on free N-glycans. These results strongly suggested that core alpha1, 6-fucose on N-glycans as well as the antennary structures of N-glycans and the polypeptide regions are required for the polysialylation by ST8Sia II. Furthermore, oligo and single alpha2, 8-sialylated glycoproteins were no longer polysialylated by mouse ST8Sia II. Therefore, the single enzyme, ST8Sia II, directly transferred all alpha2,8-sialic acid residues on the alpha2,3-linked sialic acids of N-glycans of specific NCAM isoforms to yield PSA-NCAM. Polysialylation did not require any initiator alpha2, 8 sialyltransferase but did depend on the carbohydrate and protein structures of NCAM. PMID- 8702636 TI - Retinoid X receptor alters the determination of DNA binding specificity by the P box amino acids of the thyroid hormone receptor. AB - Nuclear hormone receptors bind to hormone response elements in DNA consisting of two half-sites of 6 base pairs. The P-box amino acids of each receptor determine the identities of the central nucleotides of the half-site. 57 P-box variants of the human thyroid hormone receptor (hT3Rbeta) were used to demonstrate the relationship between P-box sequence and DNA binding specificity by homodimers and heterodimers formed with the retinoid X receptor (RXR). In general, the formation of heterodimers relieved many of the constraints on the compatibility of hT3Rbeta P-box sequences with DNA binding. Effects were most dramatic for heterodimers bound to a direct repeat spaced by four base pairs. RXR also overrides the P-box derived DNA binding specificity of hT3Rbeta when heterodimers are bound to inverted or everted repeat elements. These effects of RXR are most pronounced on AGGTCA half-sites but are squelched when the RXR partner of the heterodimer is bound to an AGGACA half-site. The influence of RXR on hT3Rbeta DNA binding specificity varies with the orientation of half-sites in the element, the identity of the fourth base pair of the half-site, and the spacing between the half-sites of direct repeats. These differences suggest that the DNA binding domains of RXR-hT3Rbeta heterodimers are not positioned equivalently on the various elements, affecting the manner in which the P-box amino acids of hT3Rbeta interact with base pairs within the half-site. PMID- 8702637 TI - Isolation of markers for chondro-osteogenic differentiation using cDNA library subtraction. Molecular cloning and characterization of a gene belonging to a novel multigene family of integral membrane proteins. AB - To identify novel marker molecules associated with chondro-osteogenic differentiation, we have set up a differential screening system based on a cDNA library subtraction in organ cultures of prenatal mouse mandibular condyles. Differential screening of a cDNA library constructed from in vitro cultured condyles allowed the isolation of a novel gene, named E25. Full-length E25 cDNA is predicted to encode a type II integral membrane protein of 263 amino acid residues. In situ hybridization experiments show that E25 is expressed in the outer perichondrial rim of the postnatal mandibular condyle, which contains the proliferating progenitor cells, but not in the deeper layers of the condyle containing the more differentiated chondroblasts and chondrocytes. Other cartilagenous tissues and their perichondrium were negative. Strong in situ hybridization signals were also detected on bone trabeculae of mature bone in tooth germs and in hair follicles. Northern blot analysis showed strong expression in osteogenic tissues, such as neonatal mouse calvaria, paws, tail, and in skin. This expression profile suggests that E25 could be a useful marker for chondro-osteogenic differentiation. Homology searches of DNA databanks showed that E25 belongs to a novel multigene family, containing three members both in man and mouse. The mouse E25 gene locus (Itm2) was mapped to the X chromosome. PMID- 8702638 TI - An epidermal growth factor receptor/Jak2 tyrosine kinase domain chimera induces tyrosine phosphorylation of Stat5 and transduces a growth signal in hematopoietic cells. AB - The Jak family of tyrosine kinases and the Stat family of transcription factors have been implicated in transducing signals from the hematopoietic growth factor receptors. To explore the role played by a member of the Jak family, Jak2, in hematopoietic cell growth signaling, we constructed a chimeric cDNA coding for the Jak2 tyrosine kinase domain linked to the extracellular and transmembrane regions of the epidermal growth factor (EGF) receptor (EGFR) and expressed the chimera in an interleukin (IL)-3-dependent cell line, 32D. When deprived of IL-3, EGF prevented apoptosis of the transfected cells, induced dose-dependent proliferation, and supported long-term growth. EGF stimulation of the transfectants induced dose-dependent tyrosine phosphorylation of the EGFR/Jak2 chimera and Stat5, which correlated with the EGF dose dependence of cell proliferation. On the other hand, EGF did not induce tyrosine phosphorylation of other factors implicated in cytokine receptor signaling, including the IL-3 receptor beta subunit, Jak kinases, Stat proteins other than Stat5, Shc, Syp, and mitogen-activated protein kinases. These results suggest that the activation of Jak2 may be sufficient for transducing a growth signal in hematopoietic cells by activating the Stat5 pathway or previously unidentified signaling pathways. In addition, because EGF induces homodimerization of the EGFR to activate its tyrosine kinase activity, the present study, which shows EGF-dependent activation of the EGFR/Jak2 chimera, implies that Jak2 may also become activated by homodimerization. PMID- 8702639 TI - Fibrillin-1 and fibulin-2 interact and are colocalized in some tissues. AB - Microfibrils 10-12 nm in diameter are found in elastic and non-elastic tissues with fibrillin as a major component. Little is known about the supramolecular structure of these microfibrils and the protein interactions it is based on. To identify protein binding ligands of fibrillin-1, we tested binding of recombinant fibrillin-1 peptides to different extracellular matrix proteins in solid phase assays. Among the proteins tested, only fibulin-2 showed significant binding to rF11, the N-terminal half of fibrillin-1, in a calcium-dependent manner. Surface plasmon resonance demonstrated high affinity binding with a Kd = 56 nM. With overlapping recombinant fibrillin-1 peptides, the binding site for fibulin-2 was narrowed down to the N terminus of fibrillin-1 (amino acid positions 45-450). Immunofluorescence in tissues demonstrated colocalization of fibrillin and fibulin-2 in skin, perichondrium, elastic intima of blood vessels, and kidney glomerulus. Fibulin-2 was not present in ocular ciliary zonules, tendon, and the connective tissue around kidney tubules and lung alveoli, which all contain fibrillin. Immunogold labeling of fibulin-2 on microfibrils in skin was found preferentially at the interface between microfibrils and the amorphous elastin core, suggesting that in vivo the interaction between fibrillin-1 and fibulin-2 is regulated by cellular expression and deposition as well as by protein-protein interactions. PMID- 8702640 TI - Role of RuvA in branch migration reactions catalyzed by the RuvA and RuvB proteins of Escherichia coli. AB - The RuvA and RuvB proteins of Escherichia coli promote ATP-dependent branch migration of Holliday junctions during homologous genetic recombination and DNA repair. In this process, RuvA acts as a specificity factor that targets RuvB, a hexameric ring motor protein, to the junction. Because elevated concentrations of RuvB can promote branch migration in the absence of RuvA, it has been suggested that RuvA acts as a molecular matchmaker. In the studies presented here, we compared the requirements for RuvAB- and RuvB-mediated branch migration reactions and found that reactions catalyzed by RuvB alone were highly sensitive to inhibition by NaCl, temperature, ADP, and ATPgammaS. Our observations indicate that the two reactions occur by distinct mechanisms and support the notion that RuvAB-mediated branch migration is physiologically more relevant than that catalyzed by RuvB. We also show that ongoing RuvAB-mediated branch migration reactions were blocked by the addition of polyclonal antibodies raised against RuvA. The role of RuvA is therefore unlikely to be restricted to RuvB targeting; instead, it is required continually during branch migration. Competition with excess synthetic Holliday junctions, sufficient to sequester released RuvA, failed to cause an immediate block and leads us to suggest that RuvAB promote branch migration by a processive mechanism. PMID- 8702641 TI - Dopamine D1A receptor regulation of phospholipase C isoform. AB - In LTK- cells stably transfected with rat D1A receptor cDNA, fenoldopam, a D1 agonist, increased phosphatidylinositol 4, 5-bisphosphate hydrolysis in a time dependent manner. In the cytosol, phospholipase C (PLC) activity increased (50 +/ 7%) in 30 s, returned to basal level at 4 h, and decreased below basal values by 24 h; in the membrane, PLC activity also increased (36 +/- 13%) in 30 s, returned to basal level at 10 min, and decreased below basal value at 4 and 24 h. Fenoldopam also increased PLC-gamma protein in a time-dependent manner. The latter was blocked by the D1 antagonist SKF83742 and by a D1A antisense oligodeoxynucleotide, indicating involvement of the D1A receptor. The fenoldopam induced increase in PLC-gamma and activity was mediated by protein kinase A (PKA) since it was blocked by the PKA antagonist Rp-8-CTP-adenosine cyclic 3':5' monophosphorothioate (Rp-8-CTP-cAMP-S) and mimicked by direct stimulation of adenylyl cyclase with forskolin or by a PKA agonist, Sp-cAMP-S. Protein kinase C (PKC) was also involved, since the fenoldopam-induced increase in PLC-gamma protein was blocked by two different PKC inhibitors, calphostin C and chelerythrine; calphostin C also blocked the fenoldopam-induced increase in PLC activity. In addition, forskolin and a PKA agonist, Sp-8-CTP-cAMP-S, increased PKC activity, and direct stimulation of PKC with phorbol 12-myristate 13-acetate increased PLC-gamma protein and activity, effects that were blocked by calphostin C. We suggest that the D1A-mediated stimulation of PLC occurs as a result of PKA activation. PKA then stimulates PLC-gamma in cytosol and membrane via activation of PKC. PMID- 8702642 TI - Efficient folding of firefly luciferase after transport into mammalian microsomes in the absence of luminal chaperones and folding catalysts. AB - Folding of polypeptides emerging from the protein translocase in the membrane of mammalian microsomes was analyzed after synthesis of corresponding precursor proteins in a mammalian translation system. Firefly luciferase was used as a model protein; the corresponding hybrid precursor contained the preprolactin signal peptide. The rates and efficiencies of folding of luciferase in microsomes were compared with those of folding of luciferase in the cytosol. Furthermore, folding of luciferase in microsomes was compared with that in proteoliposomes, i.e. in the absence of luminal molecular chaperones and folding catalysts. Folding in microsomes was less efficient compared with folding in the cytosol. Folding in the absence of luminal proteins was more efficient compared with folding in their presence and identical to folding in the cytosol. Thus, firefly luciferase emerging from translocase can efficiently fold to its native conformation without chaperoning by any luminal proteins. There may be molecular chaperones present in the microsomal membrane that can efficiently substitute for the cytosolic chaperone machinery comprising Hsp40, Hsp60, and Hsp70 with respect to folding of firefly luciferase. PMID- 8702643 TI - Cysteine string proteins are associated with chromaffin granules. AB - In this work, we have examined the subcellular distribution of cysteine string proteins (Csps) in bovine adrenal medullary chromaffin cells. Csps did not leak from digitonin-permeabilized chromaffin cells, suggesting that there is no cytosolic pool of the protein in these cells. Subcellular fractionation studies confirmed that there was essentially no Csp immunoreactivity in the cytosolic fraction. However, immunoreactivity was detected in the membrane fractions of these cells. Csp immunoreactivity codistributed with dopamine beta-hydroxylase, a granule marker protein, in sucrose gradient-separated granule fractions. Immunofluorescence studies showed that all chromaffin cells in culture were stained with a punctate appearance consistent with a granular localization. These results were confirmed by immunogold labeling, which demonstrated specific labeling of chromaffin granule membranes. In addition to its presence on synaptic vesicles, cysteine string protein is therefore a bona fide chromaffin granule membrane protein. PMID- 8702644 TI - Cell-surface expression of an amino-terminal fragment of apolipoprotein B increases lipoprotein lipase binding to cells. AB - Previous studies (Sivaram, P., Choi, S. Y., Curtiss, L. K., and Goldberg, I. J.(1994) J. Biol. Chem. 269, 9409-9412) from this laboratory showed that the NH2 terminal region of apoB (NTAB) has binding domains for lipoprotein lipase (LPL). LPL binding to endothelial cells, we hypothesize, involves interaction both with heparan sulfate proteoglycans and with a protein that has homology to NTAB. To test whether cell-surface NTAB would increase the amount and affinity of LPL binding to cells, we produced stable Chinese hamster ovary cell lines that have NTAB anchored to the cell surface. A cDNA encoding the amino-terminal 17% of apoB (apoB17) was fused to a cDNA coding for the last 37 amino acids of decay accelerating factor (DAF), which contains the signal for glycosylphosphatidylinositol anchor attachment. The fused construct was sequence verified and cloned into expression vector pCMV5. The pCMV5-apoB17-DAF plasmid was cotransfected with a neomycin resistance gene into wild-type (WT) cells and mutant heparan sulfate proteoglycan-deficient Chinese hamster ovary cells (745 cells), and stable cell lines were established. Expression of apoB17 on the cell surface was confirmed by the release of apoB17 by phosphatidylinositol-specific phospholipase C. LPL binding to WT and apoB17-DAF-transfected cells was determined. Using 0.8-6 microg of LPL, 1.3-2.2-fold more LPL associated with apoB17-DAF WT cells compared with WT cells; apoB17-DAF also increased LPL binding to 745 cells. After heparinase treatment, LPL binding to apoB17-DAF cells was still greater than to treated WT cells. This increased binding to apoB17-DAF cells was almost abolished by treatment of cells with phosphatidylinositol specific phospholipase C or anti-apoB monoclonal antibody. LPL dissociated from WT cells with k-1 = 2.55 x 10(-2) min-1, whereas LPL dissociated more slowly from apoB17-DAF-containing cells with k-1 = 1.08 x 10(-2) min-1. Furthermore, almost 95% of the LPL on WT cells was dissociated by 1 M NaCl, while only 65% of the LPL dissociated from apoB17-DAF cells at the same high salt concentration. Similarly, in high salt, more LPL remained associated with apoB17-DAF cells than with nontransfected 745 cells. These data show that NTAB on cell surfaces can function as a LPL-binding protein. Moreover, they demonstrate that LPL association with cells can be increased by simultaneously binding to both proteoglycan and non proteoglycan binding sites. PMID- 8702645 TI - Thapsigargin-induced calcium influx in the absence of detectable tyrosine phosphorylation in human platelets. AB - Tyrosine phosphorylation is a potential mechanism for mediating store-operated calcium (SOC) influx in platelets and other nonexcitable cells. Thapsigargin induces calcium-dependent tyrosine phosphorylation and SOC influx in platelets. We prevented thapsigargin-induced tyrosine phosphorylation by buffering cytosolic calcium rise with the calcium chelator 1, 2-bis-(o-aminophenoxy)ethane-N,N,N',N' tetraacetic acid-acetomethoxyester (BAPTA-AM). Calcium influx, induced by thapsigargin and measured by 45Ca2+ accumulation, persisted in BAPTA-loaded platelets in the absence of tyrosine phosphorylation. This calcium influx was blocked by the SOC influx inhibitor SKF-96365. Tyrosine kinase inhibitors have been used to demonstrate a role for tyrosine phosphorylation in SOC influx. We compared the effects of four tyrosine kinase inhibitors genistein, methyl-2, 5 dihydroxycinnamate (erbstatin analog), tyrphostin A47, and lavendustin A, on thapsigargin-induced tyrosine phosphorylation in control platelets and on thapsigargin-induced SOC influx into BAPTA-loaded platelets in absence of tyrosine phosphorylation. Tyrphostin A47 prevented all measurable tyrosine phosphorylation in control platelets, but did not decrease calcium influx into BAPTA-loaded platelets. Genistein and the erbstatin analog were poor inhibitors of tyrosine phosphorylation but decreased SOC influx into BAPTA-loaded platelets to 55.8 +/- 3% and 51.9 +/- 7.5% of control, respectively. Lavendustin A did not decrease tyrosine phosphorylation or calcium influx. Thus, thapsigargin-induced SOC influx can occur without detectable tyrosine phosphorylation and the inhibition of SOC influx by tyrosine kinase inhibitors does not correlate with their ability to prevent tyrosine phosphorylation. PMID- 8702646 TI - Accumulation of glucosylceramides in multidrug-resistant cancer cells. AB - Multidrug-resistant (MDR) tumors and cancer cell lines demonstrate a wide variety of biochemical changes. In this study we used drug-sensitive wild-type (wt) cancer cell lines and respective MDR subclones, and we demonstrate the accumulation of distinct lipids in MDR cells. These lipids were either absent or present at very low levels in drug-sensitive cells. The compounds, termed lipid-1 and lipid-2, migrated on thin-layer chromatography as a doublet. They could be radiolabeled by incubating MCF-7-AdrR (Adriamycin-resistant) breast cancer cells with [3H]serine, [3H]palmitic acid, or [3H]galactose. Utilization of these precursors by MCF-7-wt cells for synthesis of lipid-1 and -2 was minimal. Two inhibitors of sphingolipid biosynthesis, L-cycloserine and fumonisin B1, prevented the observed accumulation of the lipid compounds. An inhibitor of glucosylceramide synthesis, 1-phenyl-2-palmitoylamino-3-morpholino-1-propanol, completely abolished the formation of lipid-1 and -2 in MCF-7-AdrR cells and, to a lesser extent, inhibited the formation of lactosylceramides and gangliosides. Utilizing mass spectrometry, the multidrug resistance-associated lipids were further characterized as monoglycosylceramides of two major species that contained either 16-carbon (palmitic) or 24-carbon (lignoceric and nervonic) fatty acids. The carbohydrate head group of glycosylceramides was identified as glucose, not galactose, thus designating the accumulated lipids as glucosylceramides. Incorporation of [3H]palmitic acid into glucosylceramide was strikingly higher (8-10 times) in MCF-7-AdrR cells compared with MCF-7-wt cells. Since the rate of glucosylceramide degradation in MCF-7-AdrR cells was not attenuated, accelerated glycosphingolipid synthesis in MDR cells is suggested. Glucosylceramide also accumulated in KB-V-1, a vinblastine-resistant epidermoid carcinoma but not in KB-3-1, drug-sensitive wt cells. MDR ovarian adenocarcinoma cells (NIH:OVCAR-3) also contained elevated levels of glucosylceramide. Our results demonstrate a correlation between cellular drug resistance and alterations in glucosylceramide metabolism. PMID- 8702647 TI - Expression and characterization of inactivating and activating mutations in the human Ca2+o-sensing receptor. AB - Nearly 30 mutations have been identified to date in the coding region of the extracellular calcium-sensing receptor (CaR) that are associated with inherited human hypo- and hypercalcemic disorders. To understand the mechanisms by which the mutations alter the function of the receptor may help to discern the structure-function relationships in terms of ligand-binding and G protein coupling. In the present studies, we transiently expressed eight known CaR mutations in HEK293 cells. The effects of the mutations on extracellular calcium- and gadolinium-elicited increases in the cytosolic calcium concentration were then examined. Seven inactivating mutations, which cause familial hypocalciuric hypercalcemia and neonatal severe hyperparathyroidism, show a reduced functional activity of the receptor because they may 1) reduce its affinity for agonists; 2) prevent conversion of the receptor from a putatively immature, high mannose form into the fully glycosylated and biologically active form of the CaR, in addition to lowering its affinity for agonists; or 3) fail to couple the receptor to and/or activate its respective G protein(s). Conversely, one activating mutation, which causes a form of autosomal dominant hypocalcemia, appears to increase the affinity of the receptor for its agonists. PMID- 8702648 TI - Stoichiometry and site-specific phosphorylation of human progesterone receptor in native target cells and in the baculovirus expression system. AB - Human progesterone receptor (PR) in T47D breast cancer cells is phosphorylated on nine different serine residues; three are hormone-inducible (Ser102, Ser294, and Ser345), while others are basal but hormone-stimulated. In the present study, we have compared the phosphorylation state of native and recombinant PR expressed in a baculovirus insect cell system. Stoichiometric measurements showed that unliganded native PR in T47D cells was approximately 50% phosphorylated ( approximately 4 phosphates/PR) and became essentially 100% phosphorylated ( approximately 9 phosphates/PR) when bound to hormone. Unliganded PR expressed in Sf9 insect cells was phosphorylated with a similar stoichiometry ( approximately 3 phosphates/PR), but the phosphate content did not change with hormone addition. Site-specific phosphorylation analyzed by tryptic phosphopeptide mapping and manual peptide sequencing revealed that expressed PR bound to hormone in the Sf9 insect cells was phosphorylated on all the same sites as hormone-treated PR in T47D cells. Only minor differences were detected in the relative proportion of three sites (two basal sites and Ser345) and phosphorylation did not occur on alternate sites. Interestingly, unliganded baculovirus-expressed PR was constitutively phosphorylated on hormone inducible sites and was phosphorylated on basal sites to the same extent as hormone treated PR. Thus, in the absence of hormone, the phosphorylation state of baculovirus-expressed PR resembled that of the hyperphosphorylated native PR. In contrast to native PR, the expressed receptor in cytosols of Sf9 cells did not form a large oligomeric complex suggesting that hyperphosphorylation may be due to dissociation of the complex in the absence of hormone. This study demonstrating phosphorylation on correct sites with a stoichiometry similar to that of native PR indicates that overexpressed PR in the baculovirus system is suitable for in vitro structure/function studies. PMID- 8702649 TI - Factor-dependent release of nascent RNA by ternary complexes of vaccinia RNA polymerase. AB - Factor-dependent transcription termination during synthesis of vaccinia early mRNAs occurs at heterogeneous sites downstream of a UUUUUNU signal in the nascent transcript. The choice of termination site is flexible and is determined by a kinetic balance between nascent chain elongation and the transmission of the RNA signal to the polymerase. To eliminate ongoing elongation as a variable, we have established a system to study transcript release by purified ternary complexes halted at a defined template position 50-nucleotides 3' of the first U residue of the termination signal. Release of the nascent RNA depends on the vaccinia termination factor (VTF) and an ATP cofactor. Transcript release is blocked by BrUMP substitution within the termination signal of the nascent RNA. In these respects, the release reaction faithfully mimics the properties of the termination event. We demonstrate that ternary complexes are refractory to VTF mediated transcript release when the first U of the UUUUUNU signal is situated 20 nucleotides from the growing point of the nascent chain. Ribonuclease footprinting of the arrested ternary complexes defines a nascent RNA binding site on the polymerase elongation complex that encompasses a 16-21 nucleotide RNA segment extending proximally from the 3' end of the chain. We surmise that access of VTF to the signal sequence is prevented when UUUUUNU is bound within the nascent RNA binding site. Hence, physical not kinetic constraints determine the minimal distance between the signal and potential sites of 3' end formation. PMID- 8702650 TI - The role of the zinc motif in sequence recognition by DNA primases. AB - The DNA primase of bacteriophage T7 has a zinc-binding motif that is essential for the recognition of the sequence 3'-CTG-5'. The T7 primase also catalyzes helicase activity, a reaction coupled to nucleotide hydrolysis. We have replaced the zinc motif of the T7 primase with those found in the gene 61 primase of phage T4 and the DnaG primase of Escherichia coli. The T4 and E. coli primases recognize the sequences 3'-T(C/T)G-5' and 3'-GTC-5', respectively. Both chimeric proteins can partially replace T7 primase in vivo. The two chimeric primases catalyze the synthesis of oligoribonucleotides albeit at a reduced rate and DNA dependent dTTPase activity is reduced by 3-10-fold. Both chimeric proteins recognize 3'-(A/G)CG-5' sites on single-stranded DNA, sites that differ from those recognized by the T7, T4, or E. coli primases, indicating that the zinc motif is only one determinant in site-specific recognition. PMID- 8702651 TI - Eukaryotic expression of recombinant biglycan. Post-translational processing and the importance of secondary structure for biological activity. AB - Biglycan is a small chondroitin sulfate proteoglycan found in many tissues and is structurally related to decorin, fibromodulin, and lumican. The biological function of biglycan is poorly understood, although several studies have indicated interaction with other extracellular matrix components. We have initiated studies of structural and functional domains of biglycan by transient eukaryotic expression using the vaccinia virus/T7 bacteriophage expression system. A recombinant vaccinia virus, vBGN4 encoding the mature biglycan core protein as a polyhistidine fusion protein under control of the T7 phage promoter was expressed in HT-1080 cells and UMR106 cells. The structure of the recombinant biglycan secreted by these cells was defined by analyzing molecules labeled in the presence of [35S]sulfate, [3H]glucosamine, and [35S]methionine. Glycoforms of biglycan were separated by imidazole gradient elution, under non-denaturing conditions, and comprised: a large proteoglycan form substituted with two chondroitin sulfate chains of molecular mass approximately 34 kDa (HT-1080 cells) or approximately 40 kDa (UMR106 cells); a small proteoglycan form substituted with two chondroitin sulfate chains with a median molecular mass approximately 28 kDa; and a core protein form secreted devoid of glycosaminoglycan chains. All the glycoforms were substituted with two N-linked oligosaccharides, and the disaccharide composition of the two glycosaminoglycan populations were identical. Approximately 70% of the recombinant biglycan secreted by HT-1080 cells was substituted with chondroitin sulfate chains, whereas about 50% of the biglycan expressed by UMR106 cells was substituted with chondroitin sulfate chains. Infection with vBGN4 in both HT-1080 and UMR106 cells resulted in the production of approximately 10 mg of biglycan/10(9) cells per 24 h. The native recombinant biglycan was shown to bind to collagen type V and the complement protein, C1q. However, when the secondary structure of recombinant biglycan was disrupted by exposure to 4 M guanidine hydrochloride, the affinity for collagen type V was dramatically reduced. These data demonstrate the importance of secondary structure to the function of this small proteoglycan. PMID- 8702652 TI - Recombinant decorin glycoforms. Purification and structure. AB - The vaccinia virus/T7 bacteriophage expression system was used to express human decorin in HT-1080 cells by co-infection with vTF7-3, encoding T7 RNA polymerase, and vDCN1, encoding the decorin core protein fused to a polyhistidine-insulin signal sequence fusion-protein cassette. Overexpression using the vaccinia virus/T7 phage system resulted in secretion of approximately 30 mg of decorin/10(9) cells per 24 h which enabled purification and separation of multiple glycoforms under native conditions. Cells were cultured in the presence of [35S]methionine or a mixture of [3H]glucosamine and [35S]sulfate, and recombinant glycoprotein purified by metal affinity chromatography which resolved the secreted decorin into two classes, a proteoglycan form and a core protein form. About 25% of the recombinant protein was secreted into the culture medium as core protein devoid of glycosaminoglycan chains. The decorin core protein was resolved into two forms (approximately 49 and approximately 53 kDa) that differed in the extent of N-linked oligosaccharide substitution (2 and 3 N-linked oligosaccharides, respectively). Deglycosylation of the recombinant proteoglycans and core proteins resulted in a single band migrating with an apparent molecular mass approximately 43 kDa when analyzed by SDS-polyacrylamide gel electrophoresis. Far-UV circular dichroism spectra of native decorin proteoglycan showed a minima at 218 nm, consistent with a secondary structure that is predominantly beta-sheet. Circular dichroism spectra of bovine decorin extracted from articular cartilage and recombinant decorin similarly treated revealed a minima of 205 nm indicating a loss of secondary structure. The affinity of decorin proteoglycan and core protein for collagen-like molecules was demonstrated, with the complement component C1q exhibiting the most striking affinity for decorin, although adherence to collagen types I and V was also observed. The extensive secondary structure maintained in the purified recombinant protein is likely to be important for the biological function of decorin. PMID- 8702653 TI - An activating mutation in the ATP binding site of the ABL kinase domain. AB - A number of structural alterations have been shown to activate the leukemogenic potential of the ABL oncogene, but there is little understanding of the regulatory mechanisms that are subverted by such changes. We have used directed mutagenesis to examine a potential regulatory motif in cABL, which could directly influence ABL tyrosine kinase activity. A tyrosine to phenylalanine substitution within the ATP binding fold of the ABL kinase domain is sufficient to activate cABL enzymatic activity, and the mutant protein will alleviate growth factor dependence when expressed in the BA/F3 cell line. This growth promotion is dependent upon the structure of the amino terminus of the protein, and the ABL mutation will cooperate with certain BCR sequences in BCR/ABL fusion proteins to deregulate ABL kinase activity. PMID- 8702654 TI - Wild-type and mutant vacuolar membranes support pH-dependent reassembly of the yeast vacuolar H+-ATPase in vitro. AB - Treatment of the yeast vacuolar proton-translocating ATPase (H+-ATPase) with 300 mM KI in the presence of 5 mM MgATP results in a 90% inhibition of ATPase activity accompanied by removal of at least five of the peripheral subunits of the enzyme from the membrane. Functional reassembly of the enzyme, as indicated by reattachment of the peripheral subunits and a partial (30-70%) recovery of ATPase activity, could be achieved by dialysis of the stripped wild-type membranes to remove the KI and MgATP, but proved to be strongly pH-dependent, with optimal reassembly and recovery of activity occurring after dialysis at pH 5.5. Vacuolar membranes isolated from vma2Delta mutants, which lack one of the peripheral subunits of the enzyme, do not contain any of the peripheral subunits but are shown to contain assembled membrane (Vo) complexes. The vma2Delta mutant vacuoles are demonstrated to be competent for attachment of KI-stripped peripheral subunits and reactivation of ATPase activity. The results indicate that previously assembled Vo complexes are capable of inducing assembly of the peripheral subunits, both with each other and with the membrane subunits, and of activating the ATPase activity that resides in the peripheral subunits in a pH dependent manner. PMID- 8702655 TI - Purification of the pets factor. A nuclear protein that binds to the inducible TG rich element of the human immunodeficiency virus type 2 enhancer. AB - The peri-ets (pets) site is a TG-rich element found immediately adjacent to two binding sites for the ets family member Elf-1 in the human immunodeficiency virus type 2 (HIV-2) enhancer. Enhancer activation in response to T cell stimulation by phorbol myristate acetate, phytohemagglutinin, soluble or cross-linked antibodies to the T cell receptor, or antigen is mediated through this site in conjunction with its two adjacent Elf-1 binding sites, PuB1 and PuB2, and a kappaB site. Site specific mutation of the pets element significantly reduces inducible activation of this enhancer but does not affect its transactivation by HIV-2 tat or other viral transactivators. Similar TG-rich sequences adjacent to ets-binding sites have also been found to be functionally important in the human T-cell leukemia virus type I and murine Moloney leukemia virus enhancers. As the cellular factor binding to the pets site plays a significant role in regulating the HIV-2 enhancer in both T cells and monocytes, we have purified this protein from bovine spleens and demonstrate that it is 43 kDa in size. In addition, using glycerol gradient centrifugation, Southwestern blotting, electrophoretic mobility shift assays employing purified protein eluted from a gel, and a new in solution UV cross-linking competitive assay, we show that the dominant protein binding to the pets site is 43 kDa in size. These results indicate that a nuclear protein of 43 kDa binds specifically to the pets site of the HIV-2 enhancer and may mediate transcriptional activation of this important human pathogen in response to T cell stimulation. As retroviruses generally expropriate important human regulatory proteins for their own use, the 43-kDa pets factor is also likely to play a significant role in signal transduction in T cells and in other cellular processes. PMID- 8702656 TI - Development of the first potent and selective inhibitor of the zinc endopeptidase neurolysin using a systematic approach based on combinatorial chemistry of phosphinic peptides. AB - A new systematic approach, based on combinatorial chemistry of phosphinic peptides, is proposed for rapid development of highly potent and selective inhibitors of zinc metalloproteases. This strategy first evaluates the effects on the inhibitory potency and selectivity of the following parameters: 1) size of the phosphinic peptides, 2) position of the phosphinic bond in the sequence, and 3) the state (free or blocked) of the peptide extremities. After this selection step, the influence of the inhibitor sequence is analyzed in order to determine the identity of the residues that optimized both the potency and the selectivity. We demonstrate the efficiency of this novel approach in rapid identification of the first potent inhibitor of the mammalian zinc endopeptidase neurolysin(24-16), able to discriminate between this enzyme and the related zinc endopeptidase thimet oligopeptidase(24-15). The most potent and selective inhibitor developed in this study, Pro-LPhePsi(PO2CH2)Gly-Pro, displays a Ki value of 4 nM for 24-16 and is 2000 times less potent on 24-15. The specific recognition of such a free phosphinic tetrapeptide by 24-16, as well as the unique specificity of the 24-16 S2 and S2' subsites for proline, unveiled by this study, are discussed in terms of their possible significance for the function of this enzyme and its related zinc endopeptidase activities. PMID- 8702657 TI - Glucocorticoid-mediated repression of NFkappaB activity in endothelial cells does not involve induction of IkappaBalpha synthesis. AB - Repression of NFkappaB-dependent gene expression is one of the major elements of immunosuppression by glucocorticoids. Protein-protein interactions between the glucocorticoid receptor and NFkappaB have been characterized and shown to be a possible mechanism of mutual inhibition of transactivation properties. More recently, glucocorticoid-mediated induction of IkappaBalpha, an inhibitor of NFkappaB, has been described in monocytes and lymphocytes; an increase in IkappaBalpha mRNA and protein resulted in inactivation and cytosolic retention of NFkappaB. Thus, rather than the physical interaction between the glucocorticoid receptor and NFkappaB, the up-regulation of IkappaBalpha was presented as the key element in immunosuppression by glucocorticoids. In contrast, we show that the IkappaBalpha pathway is not involved in glucocorticoid-mediated inhibition of NFkappaB activity in endothelial cells. Although transcriptional activation by NFkappaB was significantly reduced in the presence of glucocorticoids, we did not detect induction of IkappaBalpha protein that could prevent nuclear translocation of NFkappaB upon stimulation with lipopolysaccharide or tumor necrosis factor alpha. Furthermore, treatment with glucocorticoids did not seem to affect the transcription rate or mRNA stability of IkappaBalpha. We therefore conclude that, although induction of IkappaBalpha expression by glucocorticoids seems to be of importance in monocytes and lymphocytes, it cannot explain inhibition of NFkappaB dependent gene expression in endothelial cells. Our results emphasize the relevance of physical interaction between the glucocorticoid receptor and NFkappaB in endothelial cells and thus in suppression of inflammation by glucocorticoids. PMID- 8702658 TI - Regulation of the heat-shock protein 70 reaction cycle by the mammalian DnaJ homolog, Hsp40. AB - The effects of the human DnaJ homolog, Hsp40, on the ATPase and chaperone functions of the constitutively expressed Hsp70 homolog, Hsc70, were analyzed. Hsp40 stimulates the hydrolysis of ATP by Hsc70, causing a approximately 7-fold increase in its steady-state ATPase activity. In contrast to the prokaryotic Hsp70 system, ATP-hydrolysis and not the release of bound ADP is the rate limiting step in the overall ATPase cycle of mammalian Hsc70. The ability to activate the Hsc70 ATPase is partially preserved in a deletion mutant containing the J-domain and the G/F region of Hsp40 but not in a deletion mutant that contains the J-domain alone. As a result of its ATPase stimulating activity, addition of Hsp40 allows Hsc70 to bind peptide in the presence of ATP, whereas in the absence of Hsp40, peptide is efficiently released upon ATP binding to Hsc70. The functional cooperation of Hsp40 with Hsc70 is essential to ensure the ATP hydrolysis-dependent binding of aggregation-sensitive denatured polypeptides, such as thermally denatured firefly luciferase and chemically denatured rhodanese. Binding of these proteins results in the formation of ternary complexes of Hsc70, Hsp40, and substrates. Hsc70 and Hsp40 cooperate with further factors in protein renaturation, as demonstrated by the finding that luciferase, thermally denatured in the presence of Hsc70, Hsp40, and ATP, refolds upon addition of rabbit reticulocyte cytosol. Our results indicate that Hsp40 has a critical regulatory function in the Hsc70 ATPase cycle that is required for the efficient loading of peptide substrate onto Hsc70. PMID- 8702659 TI - The ATP-activated hexameric helicase of bacteriophage T4 (gp41) forms a stable primosome with a single subunit of T4-coded primase (gp61). AB - We have examined the formation of the primosome subassembly of the bacteriophage T4-coded DNA replication (elongation) complex from its helicase, primase, and DNA components. Previously, we had shown that the T4 helicase (gp41) exists in solution in a stable monomer left and right arrow dimer equilibrium at physiological protein (and salt) concentrations and forms a hexamer upon activation by ATP (or GTP) binding (Dong, F., Gogol, E. P., and von Hippel, P. H.(1995) J. Biol. Chem. 270, 7462-7473). Here we report that the T4 primase (gp61) is a monomer in solution under the same conditions, and that the ATP activated helicase binds to a single gp61 primase molecule on appropriate DNA templates to reconstitute a stable primosome. We show that: (i) the gp41 helicase alone does not form a stable complex with DNA templates, although this helicase by itself can carry out moderately processive ATP-driven translocation along single-stranded DNA (Young, M. C., Schultz, D. E., Ring, D., and von Hippel, P. H.(1994) J. Mol. Biol. 235, 1447-1458); (ii) the primase alone does form a stable complex with DNA; (iii) the helicase can bind to the primase-DNA complex in the presence of ATP or GTP to form a stable ternary complex; (iv) this complex consists of six helicase subunits and one primase subunit; and (v) the reconstituted primosome is stable for at least 10 to 20 min after NTP cleavage and dissociation of the hydrolysis products. These results strongly suggest that the functional T4 DNA replication primosome consists of an integrated 6:1 helicase-primase complex bound to DNA, and that the ATP-activated helicase hexamer remains intact throughout the processive DNA replication process. PMID- 8702660 TI - Retroviral integrase, putting the pieces together. PMID- 8702661 TI - Cells arrested in G1 by the v-Abl tyrosine kinase do not express cyclin A despite the hyperphosphorylation of RB. AB - The v-Abl tyrosine kinase encoded by the Abelson murine leukemia virus (A-MuLV) can either stimulate or inhibit cell proliferation, depending on the cell context. In a NIH-3T3-derived cell line, N3T3, v-Abl blocks the serum-induced entry into S phase. In these G1-arrested cells v-Abl does not interfere with the activation of cyclin D1 or cyclin E-dependent kinases. As a result, v-Abl does not block the hyperphosphorylation and inactivation of the retinoblastoma protein RB. However, activation of cyclin A-dependent kinase is inhibited due to a v-Abl induced block in the accumulation of cyclin A mRNA and protein. Ectopic expression of cyclin A enabled the v-Abl-arrested cells to enter S phase, whereas cyclins E and D1, or E2Fs 1 and 4 could not overcome the v-Abl arrest. Taken together, these results suggest that v-Abl tyrosine kinase arrests cell cycle progression in G1 by inhibiting the expression of cyclin A. PMID- 8702662 TI - Phosphorylation of SLP-76 by the ZAP-70 protein-tyrosine kinase is required for T cell receptor function. AB - Two families of tyrosine kinases, the Src and Syk families, are required for T cell receptor activation. While the Src kinases are responsible for phosphorylation of receptor-encoded signaling motifs and for up-regulation of ZAP 70 activity, the downstream substrates of ZAP-70 are unknown. Evidence is presented herein that the Src homology 2 (SH2) domain-containing leukocyte protein of 76 kDa (SLP-76) is a substrate of ZAP-70. Phosphorylation of SLP-76 is diminished in T cells that express a catalytically inactive ZAP-70. Moreover, SLP 76 is preferentially phosphorylated by ZAP-70 in vitro and in heterologous cellular systems. In T cells, overexpression of wild-type SLP-76 results in a hyperactive receptor, while expression of a SLP-76 molecule that is unable to be tyrosine-phosphorylated attenuates receptor function. In addition, the SH2 domain of SLP-76 is required for T-cell receptor function, although its role is independent of the ability of SLP-76 to undergo tyrosine phosphorylation. As SLP 76 interacts with both Grb2 and phospholipase C-gamma1, these data indicate that phosphorylation of SLP-76 by ZAP-70 provides an important functional link between the T-cell receptor and activation of ras and calcium pathways. PMID- 8702663 TI - Cisplatin and adriamycin resistance are associated with MutLalpha and mismatch repair deficiency in an ovarian tumor cell line. AB - In contrast to parental A2780 ovarian tumor cells, extracts of one doxorubicin resistant and two independent cis-diamminedichloroplatinum(II)-resistant derivatives are defective in strand-specific mismatch repair. The repair defect of the three hypermutable, drug-resistant cell lines is only evident when the strand break that directs the reaction is located 3' to the mismatch, and in each case repair is restored to extracts by addition of purified MutLalpha heterodimer. As judged by immunological assay, drug resistance is associated with the virtual absence of the MutLalpha MLH1 subunit and greatly reduced levels of the PMS2 subunit. These findings implicate a functional mismatch repair system in the cytotoxic effects of these antitumor drugs and may have ramifications for their clinical application. PMID- 8702664 TI - Generation and characterization of mutant mice lacking ryanodine receptor type 3. AB - The ryanodine receptor type 3 (RyR-3) functions as a Ca2+-induced Ca2+ release (CICR) channel and is distributed in a wide variety of cell types including skeletal muscle and smooth muscle cells, neurons, and certain non-excitable cells. However, the physiological roles of RyR-3 are totally unclear. To gain an insight into the function of RyR-3 in vivo, we have generated mice lacking RyR-3 by means of the gene targeting technique. The mutant mice thus obtained showed apparently normal growth and reproduction. Although Ca2+-induced Ca2+ release from intracellular Ca2+ stores of the mutant skeletal muscle differed in Ca2+ sensitivity from that of wild-type muscle, excitation-contraction coupling of the mutant muscle seemed to be normal. Moreover, we could not find any significant disturbance in the smooth muscle and lymphocytes from the mutant mice. On the other hand, the mutant mice showed increased locomotor activity, which was about 2-fold greater than that of the control mice. These results indicate that the loss of RyR-3 causes no gross abnormalities and suggest that the lack of RyR-3 mediated Ca2+ signaling results in abnormalities of certain neurons in the central nervous system. PMID- 8702665 TI - Pseudohypoparathyroidism, a novel mutation in the betagamma-contact region of Gsalpha impairs receptor stimulation. AB - Pseudohypoparathyroidism, type Ia (PHP-Ia), is a dominantly inherited endocrine disorder characterized by resistance to hormones that act by stimulating adenylyl cyclase. It is caused by inheritance of an autosomal mutation that inactivates the alpha subunit (alphas) of Gs, the stimulatory regulator of adenylyl cyclase. In three members of a family, the PHP-Ia phenotype is associated with a mutation (R231H) that substitutes histidine for an arginine at position 231 in alphas. We assessed signaling function of alphas-WT versus alphas-R231H transiently transfected in HEK293 cells. Hormone receptor-dependent stimulation of cAMP accumulation in cells expressing alphas-R231H is reduced by approximately 75% in comparison to cAMP accumulation in cells expressing alphas-WT. A second mutation, alphas-R201C, inhibits the GTPase turnoff reaction of alphas, thus producing receptor-independent stimulation of cAMP accumulation. The double mutant, alphas R231H/R201C, stimulates cAMP accumulation almost as well (approximately 80%) as does alphas-R201C itself, indicating that the R231H mutation selectively impairs receptor-dependent signaling. In three-dimensional structures of G protein heterotrimers, Arg-231 is located in a region, switch 2, that is thought to interact with the betagamma subunit rather than with the hormone receptor. Thus, the R231H phenotype suggests that switch 2 (perhaps in concert with betagamma) mediates G protein activation by receptors at a site distant from the receptor-G protein contact surface. PMID- 8702666 TI - Identification of residues of the epidermal growth factor receptor proximal to residue 45 of bound epidermal growth factor. AB - A triple mutant of murine epidermal growth factor (mEGF), N1Q/H22Y/R45K-mEGF, was constructed by site-directed mutagenesis, expressed, purified, and characterized for use in an affinity cross-linking study to identify aminoacyl residues of the EGF receptor adjacent to a residue in the carboxyl-terminal domain of bound EGF thought to be important in distinguishing between EGF and transforming growth factor-alpha in their recognition by the receptor. Cyclization of Gln1 to form pyroglutamate (pE) limited the site of cross-linking in the mutant to Lys45, permitting identification of receptor residues that are proximal to this residue of bound EGF. The resulting N1pE/H22Y/R45K-mEGF was shown to be comparable to wild-type mEGF in receptor binding and stimulation of receptor autophosphorylation. 125I-Labeled N1pE/H22Y/R45K-mEGF was reacted with the heterobifunctional cross-linking reagent sulfo-N-succinimidyl-4 (fluorosulfonyl)benzoate, and the resulting modified EGF was incubated with A431 membrane vesicles bearing EGF receptors. Incubation resulted in specific cross linking of the labeled N1pE/H22Y/R45K-mEGF to EGF receptors. The resulting cross linked complex was then partially purified, denatured, reduced, and carboxyamidomethylated. Digestion with endoprotease LysC resulted in a unique radiolabeled peptide that could be immunoprecipitated using antibodies to mEGF. This immunoprecipitated fragment was purified by gel electrophoresis and subjected to microsequencing. The resulting sequence was matched to that of a LysC fragment of the receptor, which begins with Thr464 and is near the interface of receptor subdomains III and IV. Loss of signal at cycle 2 suggests that the point of attachment of cross-linked N1pE/H22Y/R45K is Lys465 of the receptor. PMID- 8702667 TI - End-joining of free radical-mediated DNA double-strand breaks in vitro is blocked by the kinase inhibitor wortmannin at a step preceding removal of damaged 3' termini. AB - Both mammalian cells and Xenopus eggs possess activities for the joining of nonhomologous DNA ends, and such activities may play a major role in double strand break repair. In order to dissect the biochemical processing of breaks with oxidatively modified ends, vectors containing various site-specific double strand breaks with 3'-phosphoglycolate termini were constructed and treated with Xenopus egg extracts. These vectors were rejoined by the extracts at rates 30-100 times slower than comparable 3'-hydroxyl vectors. Vectors with blunt or cohesive 3'-phosphoglycolate ends yielded single repair products corresponding to simple phosphoglycolate removal followed by ligation, while a vector with mismatched ends was also rejoined but yielded a mixture of products. Addition of the kinase inhibitors wortmannin and dimethylaminopurine not only blocked rejoining, but also suppressed phosphoglycolate removal, implying an early, essential, kinase dependent restriction point in the pathway. The results suggest that double strand breaks with oxidatively modified ends are repaired in Xenopus eggs by a highly conservative and stringently regulated end-joining pathway, in which all biochemical processing of the breaks is contingent on both end alignment and a specific phosphorylation event. Several lines of indirect evidence suggest DNA dependent protein kinase as a likely candidate for effecting this phosphorylation. PMID- 8702668 TI - Insulin activates a p21-activated kinase in muscle cells via phosphatidylinositol 3-kinase. AB - Insulin activates rapidly a complex cascade of lipid and protein kinases leading to stimulation of mitogenic and metabolic events. Here we describe a renaturable kinase of 65 kDa (PK65) that becomes rapidly activated by insulin in differentiated L6 muscle cells (myotubes) and can phosphorylate histones immobilized in polyacrylamide gels. Insulin activation of PK65 was abolished by the tyrosine kinase inhibitor erbstatin and by the phosphatidylinositol 3-kinase (PI 3-kinase) inhibitor wortmannin, but was unaffected by inhibitors of protein kinase C or of the activation of p70(S6K). Recently, a number of protein kinases have been described which become activated through interaction with the small GTP binding proteins Rac and Cdc42 (21-ctivated inases, or PAKs) and lead to activation of the stress-induced mitogen-activated protein kinase (MAPK) p38 MAPK. Two different polyclonal antibodies recognizing the carboxyl-terminal or the Rac-binding domain of a 65-kDa PAK (PAK65) immunoprecipitated the myotube PK65. The insulin-induced activation of PK65 in myotubes was detectable following immunoprecipitation of the kinase. Furthermore, PK65 associated with and became activated by glutathione S-transferase-Cdc42Hs in the presence of GTPgammaS (guanosine 5'-3-O-(thio)triphosphate). In myotubes insulin also induced tyrosine phosphorylation of p38 MAPK. However, this phosphorylation was insensitive to wortmannin, indicating that p38 MAPK is not activated by PK65 in insulin stimulated cells. The results suggest that insulin activates in muscle cells a renaturable kinase (PK65) closely related to PAK65. Tyrosine kinases and PI 3 kinase act upstream of PK65 in the insulin signaling cascade. Insulin activates p38 MAPK in myotubes, but this occurs by a pathway independent of PI 3-kinase and PK65. PMID- 8702669 TI - Identification of elements of the peptide binding site of DnaK by peptide cross linking. AB - We used photocross-linking of peptides to DnaK to identify elements of the peptide binding site of DnaK. We attached a photoactivatable group (N hydroxysuccinimidyl-4-azido-salicylic acid (NHS-ASA) or N-iodoacetamidobutyl-4 azido-salicylic acid (I-ABASA)) to different positions on peptide C of the vesicular stomatitis virus glycoprotein, 125I-radiolabeled the cross-linker, cross-linked the peptide to DnaK by UV irradiation, and then determined the amino acid residues of DnaK that were cross-linked to the peptide. Limited trypsin digestion of the DnaK-peptide complex revealed that the derivatives modified with photoactivatable cross-linker peptide C cross-linked to a C-terminal fragment of DnaK and that the N-terminal 45-kDa fragment of DnaK was not cross-linked by these modified peptides. The attachment points of the three peptide C derivatives carrying photoactivatable cross-linkers at different locations on the peptide, PepC-ASA, PepC-S7C-ABASA, and PepC-S8C-ABASA, have been identified as Arg-536, Arg-527, and His-541 of DnaK, respectively. Thus all three peptides cross-linked to amino acids located close together in a sequence that includes one end of the long alpha-helix in the NMR-based secondary structure model of the peptide binding domain of Hsp70 family (Morshauser, R., Wang, H., Flynn, G., and Zuiderweg, E. (1995) Biochemistry 34, 6261-6266). PMID- 8702670 TI - Strandedness discrimination in peptide-polynucleotide complexes. AB - Preferential binding to single- or double-stranded nucleic acids is important for the activity of many proteins that process RNA and DNA. We have investigated the mechanism of strandedness discrimination with peptides derived from the putative DNA-binding domain of the RecA protein, a bacterial recombinase that modulates its affinity for single-stranded DNA by means of ATP binding and hydrolysis. Contributions of electrostatic and non-electrostatic interactions to binding of these peptides with polynucleotides were evaluated by fluorescence spectroscopy as a function of salt concentration and peptide charge. Binding of these peptides to single- and double-stranded nucleic acids was dominated by non-electrostatic interactions. Small electrostatic contributions selectively enhanced peptide complexation with single-stranded nucleic acids. Similar results were observed in control experiments carried out with tripeptides containing charged and aromatic amino acid residues. It was possible to modify the strandedness preference of peptide-polynucleotide complexes by changing electrostatic contributions to the binding free energy. These observations suggest a mechanism whereby some proteins that interact with DNA or RNA might determine and regulate their relative affinity for single- and double-stranded nucleic acids. PMID- 8702671 TI - Identification of signal-induced IkappaB-alpha kinases in human endothelial cells. AB - Activation of the nuclear transcription factor-kappaB is an early event in endothelial activation. NF-kappaB activation is regulated by the inducible phosphorylation and subsequent degradation of the inhibitory subunit IkappaB alpha. We identified two discrete kinases of approximately 36 and 41 kDa in the cytoplasm of human umbilical vein endothelial cells that specifically bind to and phosphorylate the IkappaB-alpha subunit. IkappaB-alpha kinase activity is transiently elevated following treatment with either tumor necrosis factor alpha, interleukin-1beta, or bacterial lipopolysaccharides and precedes activation of either mitogen-activated kinase or Jun kinase. Furthermore, activation of the IkappaB-alpha kinases precedes both the appearance of hyperphosphorylated IkappaB alpha and its subsequent degradation, as well as the translocation of NF-kappaB to the nucleus. Deletion mutagenesis of the IkappaB-alpha polypeptide revealed that these kinases bind in or around the ankyrin repeat domains and phosphorylate residues within the C terminus. These kinases, however, were not identical to casein kinase II and displayed a pharmacologic profile distinct from other known kinases. These kinases may represent components of a signal transduction pathway regulating IkappaB-alpha levels in vascular endothelium. PMID- 8702672 TI - Structural requirements for targeting of surfactant protein B (SP-B) to secretory granules in vitro and in vivo. AB - Human surfactant protein B (SP-B) is synthesized by type II cells as a 381 residue preproprotein which is proteolytically processed to a 79-residue mature peptide and targeted to lamellar bodies for secretion. To identify secretory granule targeting determinants, constructs encoding the SP-B preproprotein (SP B), COOH-terminally deleted SP-B (SP-BDeltaC), the NH2-terminal propeptide (SP BN), and a chimeric molecule consisting of albumin and the mature peptide (ALB/SP BM) were transfected into AtT-20 and PC12 cells. Pulse-chase studies demonstrated that 10-30% of SP-B and SP-BDeltaC remained in cells in an endoglycosidase H resistant form. Secretion of stored SP-B was stimulated by forskolin/12-O tetradecanoylphorbol-13-acetate and intracellular SP-B was localized to secretory granules by immunoelectron microscopy. In contrast, SP-BN and ALB/SP-BM were constitutively secreted and not detected in secretory granules. Specific processing of SP-B was not detected in either AtT-20 or PC12 cells. Expression of SP-BDeltaC in transgenic mice resulted in secretion of fully processed mature SP B, indicating correct processing and targeting of this construct in vivo. We conclude that 1) SP-B processing occurs in a cell-specific manner, 2) the proprotein contains secretory granule targeting determinants that are not cell specific, 3) the NH2-terminal propeptide and the mature peptide are required for targeting SP-B to lamellar body, and 4) the COOH-terminal propeptide is not required for processing or sorting of SP-B. PMID- 8702673 TI - Novel role for Sp1 in phorbol ester enhancement of human platelet thromboxane receptor gene expression. AB - Expression of platelet thromboxane receptors is transcriptionally increased during megakaryocytic differentiation stimulated by phorbol 12-myristate 13 acetate (PMA). We previously cloned and characterized the promoter region of the human thromboxane receptor gene and localized PMA-responsive elements to a region between 1.84 and 1.95 kilobase pairs (kb) 5' of the transcription initiation site (D'Angelo, D. D., Davis, M. G., Houser, W. A., Eubank, J. J., Ritchie, M. E., and Dorn, G. W., II (1995) Circ. Res. 77, 466-474). Herein we report the localization of the PMA response element to a 14-nucleotide C-rich sequence, flanked by an octanucleotide inverted repeat, located -1.938 to -1.925 kb 5' of the transcription start site of this gene. We further identify the PMA-responsive enhancer factor that binds to this C-rich sequence as Sp1. Heterologous thromboxane receptor gene promoter/thymidilate kinase reporter constructs transfected into K562 cells exhibited PMA responsiveness when the C-rich element was included with additional 3' sequence from -1.924 to -1.84 kb. However, mutations of the C-rich element that disrupted a GC box located on the inverse strand eliminated PMA responsiveness and, in gel mobility shift assays, eliminated binding of Sp1. PMA treatment of K562 cells significantly increased, by 5-fold, Sp1 binding to the C-rich element and increased both phosphorylated and nonphosphorylated Sp1 protein levels by 2-fold. Furthermore, PMA treatment transiently increased Sp1 mRNA levels prior to increasing thromboxane receptor mRNA, suggesting that up-regulation of Sp1 contributes to up-regulation of thromboxane receptors. Finally, we have detected an unidentified K562 nuclear protein that binds specifically to the sense strand of the C-rich sequence overlapping the Sp1 binding site and that, by stabilizing a double stem-loop conformation of this DNA segment, may also play a role in Sp1 regulation of this gene. These studies are the first to describe regulatory and regulated roles for Sp1 in PMA-responsive gene expression and suggest that modulation of Sp1 levels controls thromboxane receptor expression during megakaryocytic differentiation. PMID- 8702674 TI - Differential activation of mitogen-activated protein kinases by nitric oxide related species. AB - Many studies have identified nitric oxide (NO) and related chemical species (NOx) as having critical roles in neurotransmission, vasoregulation, and cellular signaling. Previous work in this laboratory has focused on elucidating the mechanism of NOx signaling in cells. We have demonstrated that NOx-induced activation of the guanine nucleotide-binding protein p21(ras) leads to nuclear translocation of the transcription factor NFkappaB. Here, we investigated whether intermediary signaling elements, namely the mitogen-activated protein (MAP) kinases, are involved in mediating NOx signaling. We found that NOx activates the extracellular signal-regulated kinase (ERK), p38, and c-Jun NH2-terminal kinase (JNK) subgroups of MAP kinases in human Jurkat T cells. JNK was found to be 100 fold more sensitive to NOx stimulation than p38 and ERK. In addition, the activation of JNK and p38 by NOx was more rapid than ERK activation. Depletion of intracellular glutathione augmented the NOx-induced increase in kinase activity. Furthermore, endogenous NO, generated from NO synthase, activated ERK, and NOx induced MAP kinase activation was effectively blocked by the farnesyl transferase inhibitor alpha-hydroxyfarnesylphosphonic acid. These data support the hypothesis that critical signaling kinases, such as ERK, p38, and JNK, are activated by NO related species and thus participate in NO signal transduction. These findings establish a role for multiple MAP kinase signaling pathways in the cellular response to NOx. PMID- 8702675 TI - Post-translational modifications of Ras and Ral are important for the action of Ral GDP dissociation stimulator. AB - Ral GDP dissociation stimulator (RalGDS) is a GDP/GTP exchange protein of Ral and a new effector protein of Ras. Therefore, there may be a new signaling pathway from Ras to Ral. In this paper, we examined the roles of the post-translational modifications of Ras and Ral on this new signal transduction pathway. The post translationally modified form of Ras bound to RalGDS more effectively than the unmodified form. The modification of Ras was required to regulate the distribution of RalGDS between the cytosol and membrane fractions in COS cells. The post-translational modification of Ral enhanced the activities of RalGDS to stimulate the dissociation of GDP from and the binding of GTP to Ral. Furthermore, the modified form of Ral bound to Ral-binding protein 1 (RalBP1), a putative effector protein of Ral, more effectively than the unmodified form. Taken together with the observations that Ras and Ral are localized to the membranes, these results suggest that the post-translational modifications of Ras and Ral play a role for transmitting the signal effectively on the membranes in the signal transduction pathway of Ras/RalGDS/Ral/RalBP1. PMID- 8702676 TI - Temperature dependence of the mitochondrial inner membrane anion channel. The relationship between temperature and inhibition by protons. AB - In this paper, we investigate the temperature and pH dependence of the mitochondrial inner membrane anion channel (IMAC) that is believed to be involved in mitochondrial volume homeostasis. At pH 7. 4, the flux of malonate is highly temperature-dependent with rates increasing from 1 nmol/min mg at 5 degrees C to 1900 nmol/min mg at 45 degrees C. The Arrhenius plot is nonlinear with the activation energy increasing from 21 kJ/mol (Q10 = 1.3) to 193 kJ/mol (Q10 = 13) as the temperature is decreased. This temperature dependence is unusual and not seen with solutes that are transported through the bilayer such as NH4OAc, malonamide, and KSCN (plus valinomycin) or even for cytochrome c oxidase dependent uptake of potassium (plus valinomycin). The temperature dependence of IMAC is closely related to the inhibition of IMAC by protons. Thus, we find that the pIC50 for protons decreases from 9.3 (Hill coefficient = 1.0) at 5 degrees C to 7.1 (Hill coefficient = 2.5) at 45 degrees C. This behavior is explained on the basis of a new kinetic model for IMAC in which the net open probability is not only modulated by the binding of three protons but also by temperature via effects on the open probability of the unprotonated channel and the pK of one of the inhibitory protonation sites. PMID- 8702677 TI - Isolation and chromosomal distribution of natural Z-DNA-forming sequences in Halobacterium halobium. AB - Conditions favoring left-handed Z-DNA such as high salinity (> 4 ), high negative DNA supercoiling, and GC-rich DNA [statistically favoring d(CG)n repeat sequences], are all found in the extremely halophilic archaeum (archaebacterium) Halobacterium halobium. In order to identify and study Z-DNA regions of the H. halobium genome, an affinity chromatography method with high Z-DNA selection efficiency was developed. Supercoiled plasmids were incubated with a Z-DNA specific antibody (Z22) and passed over a protein A-agarose column, and the bound plasmids were eluted using an ethidium bromide gradient. In control experiments using mixtures of pUC12 (Z-negative) and a d(CG)5-containing (Z-positive) pUC12 derivative, up to 4,000-fold enrichment of the Z-DNA-containing plasmid was demonstrated per cycle of the Z-DNA selection procedure. The selection efficiency was determined by transformation of Escherichia coli DH5alpha with eluted plasmids and blue-white screening on X-gal plates. Twenty recombinant plasmids containing Z-DNA-forming sequences of H. halobium were isolated from a genomic library using affinity chromatography. Z-DNA-forming sequences in selected plasmids were identified by bandshift and antibody footprinting assays using Z22 monoclonal antibody. Alternating purine-pyrimidine sequences ranging from 8 base pairs (bp) to 13 bp with at least a 6-bp alternating d(GC) stretch were found in the Z22 antibody binding regions of isolated plasmids. The distribution of Z-DNA forming sequences in the Halobacterium salinarum GRB chromosome was analyzed by dot-blot hybridization of an ordered cosmid library using the cloned H. halobium Z-DNA segments as probe. Among the 11 Z-DNA segments tested, five were found to be clustered in a 100-kilobase pair region of the genome, whereas six others were distributed throughout the rest of the genome. PMID- 8702678 TI - Identification of natural monomeric response elements of the nuclear receptor RZR/ROR. They also bind COUP-TF homodimers. AB - The receptor RZR/ROR is an important member of the nuclear receptor superfamily and has recently been shown to be the nuclear receptor for the pineal gland hormone melatonin. RZR/ROR binds as a monomer to DNA, and the human 5 lipoxygenase gene has been identified as the first RZR/ROR/melatonin-responding gene. Another prominent nuclear receptor is COUP-TF, which binds as a dimer to DNA. In this study, the sequences of known promoter regions of genes that may be involved in the physiological action of melatonin have been screened for putative monomeric RZR/ROR response elements. The binding of RZR/ROR and COUP-TF was compared and quantified on a set of 12 putative response elements. Interestingly, COUP-TF homodimers were found to bind with high affinity to some of the monomeric RZR/ROR response elements. Four RZR/ROR response elements, found in the genes of the mouse bifunctional enzyme, the rat bone sialoprotein, mouse Purkinje cell protein 2, and human p21(WAF1/CIP1), were shown to be inducible by melatonin under conditions of low constitutive activity. Surprisingly, the constitutive activity of COUP-TF was also stimulated by an unknown serum compound. The novel Purkinje cell protein 2 and p21(WAF1/CIP1) RZR/ROR/melatonin-responding genes may be the key for understanding the role of RZR/RORalpha in the mouse mutation staggerer and the antiproliferative action of melatonin, respectively. PMID- 8702679 TI - Conformational properties and stability of tyrosine hydroxylase studied by infrared spectroscopy. Effect of iron/catecholamine binding and phosphorylation. AB - The conformation and stability of recombinant tetrameric human tyrosine hydroxylase isoenzyme 1 (hTH1) was studied by infrared spectroscopy and by limited tryptic proteolysis. Its secondary structure was estimated to be 42% alpha-helix, 35% beta-extended structures (including beta-sheet), 14% beta-turns, and 10% nonstructured conformations. Addition of Fe(II) or Fe(II) plus dopamine to the apoenzyme did not significantly modify its secondary structure. However, an increased thermal stability and resistance to proteolysis, as well as a decreased cooperativity in the thermal denaturation transition, was observed for the ligand-bound forms. Thus, as compared with the apoenzyme, the ligand-bound subunits of hTH1 showed a more compact tertiary structure but weaker intersubunit contacts within the protein tetramer. Phosphorylation of the apoenzyme by cyclic AMP-dependent protein kinase did not change its overall conformation but allowed on iron binding a conformational change characterized by an increase (about 10%) in alpha-helix and protein stability. Our results suggest that the conformational events involved in TH inhibition by catecholamines are mainly related to modifications of tertiary and quaternary structural features. However, the combined effect of iron binding and phosphorylation, which activates the enzyme, also involves modifications of the protein secondary structure. PMID- 8702680 TI - Purification and cloning of the GTP cyclohydrolase I feedback regulatory protein, GFRP. AB - The activity of GTP cyclohydrolase I, the initial enzyme of the de novo pathway for biosynthesis of tetrahydrobiopterin, the cofactor required for aromatic amino acid hydroxylations and nitric oxide synthesis, is sensitive to end-product feedback inhibition by tetrahydrobiopterin. This inhibition by tetrahydrobiopterin is mediated by the GTP cyclohydrolase I feedback regulatory protein GFRP, previously named p35 (Harada, T., Kagamiyama, H., and Hatakeyama, K. (1993) Science 260, 1507-1510), and -phenylalanine specifically reverses the tetrahydrobiopterin-dependent inhibition. As a first step in the investigation of the physiological role of this unique mechanism of regulation, a convenient procedure has been developed to co-purify to homogeneity both GTP cyclohydrolase I and GFRP from rat liver. GTP cyclohydrolase I and GFRP exist in a complex which can be bound to a GTP-affinity column from which GTP cyclohydrolase I and GFRP are separately and selectively eluted. GFRP is dissociated from the GTP agarose bound complex with 0.2 NaCl, a concentration of salt which also effectively blocks the tetrahydrobiopterin-dependent inhibitory activity of GFRP. GTP cyclohydrolase I is then eluted from the GTP-agarose column with GTP. Both GFRP and GTP cyclohydrolase I were then purified separately to near homogeneity by sequential high performance anion exchange and gel filtration chromatography. GFRP was found to have a native molecular mass of 20 kDa and consist of a homodimer of 9.5-kDa subunits. Based on peptide sequences obtained from purified GFRP, oligonucleotides were synthesized and used to clone a cDNA from a rat liver cDNA library by polymerase chain reaction-based methods. The cDNA contained an open reading frame that encoded a novel protein of 84 amino acids (calculated molecular mass 9665 daltons). This protein when expressed in Escherichia coli as a thioredoxin fusion protein had tetrahydrobiopterin-dependent GTP cyclohydrolase I inhibitory activity. Northern blot analysis indicated the presence of an 0.8 kilobase GFRP mRNA in most rat tissues, the amounts generally correlating with levels of GTP cyclohydrolase I and tetrahydrobiopterin. Thus, mRNA levels were relatively high in liver and kidney and somewhat lower in testis, heart, brain, and lung. These results suggest that GFRP is widely expressed and may play a role in regulating not only phenylalanine metabolism in the liver, but also the production of biogenic amine neurotransmitters as well as nitric oxide synthesis. PMID- 8702681 TI - Induction of acetylcholine receptor gene expression by ARIA requires activation of mitogen-activated protein kinase. AB - Transcription of genes encoding nicotinic acetylcholine receptor (AChR) subunits (alpha, beta, gamma or epsilon, and delta) is highest in nuclei localized to the synaptic region of the muscle, which contributes to maintain a high density of AChRs at the postjunctional membrane. ARIA (AChR inducing activity) is believed to be the trophic factor utilized by motor neurons to stimulate AChR synthesis in the subsynaptic area. To elucidate the signaling mechanism initiated by ARIA, we established stable C2C12 cell lines carrying the nuclear lacZ gene under the control of the mouse epsilon subunit promoter or chicken alpha subunit promoter. ARIA stimulated tyrosine phosphorylation of erbB proteins in these C2C12 cells within 15 s with a peak at 5 min. Immediately following tyrosine phosphorylation of erbB proteins, mitogen-activated protein (MAP) kinase was activated which occurred within 30 s and peaked at 8 min after ARIA stimulation. Concomitantly, expression of AChR genes was induced by ARIA. ARIA-induced AChR subunit transgene expression was observed only in differentiated myotubes and not in myoblasts, suggesting that downstream signaling component(s) are regulated in a manner dependent on the myogenic program. Inhibition of the MAP kinase activity by using a specific MAP kinase kinase inhibitor or by overexpressing dominant negative mutants of Raf or MAP kinase kinase attenuated or abolished the ARIA-induced activation of AChR alpha and epsilon subunit gene expression. These results indicate that regulation of AChR gene expression by ARIA in C2C12 cells requires activation of the MAP kinase signaling pathway. PMID- 8702682 TI - Protein phosphatase 2A is essential for the activation of Ca2+-activated K+ currents by cGMP-dependent protein kinase in tracheal smooth muscle and Chinese hamster ovary cells. AB - The regulation of Ca2+-activated K+ channels (KCa channels) by cGMP-dependent protein kinase (cGMP kinase) and its molecular mechanism were investigated in Chinese hamster ovary (CHO) and tracheal smooth muscle cells. In CHO wild-type cells (CHO-WT cells) and in CHO cells stably transfected with cGMP kinase Ialpha (CHO-cGK cells), KCa channels with intermediate conductance (approximately 50 picosiemens) were identified. Due to the basal activity of cGMP kinase, Ca2+ activated K+ currents had a higher sensitivity toward the cytosolic Ca2+ concentration in CHO-cGK cells than in CHO-WT cells. Dialysis of the active fragment of cGMP kinase (300 n) into CHO-WT cells or of cGMP into CHO-cGK cells increased the Ca2+-activated K+ current, while the catalytic subunit of cAMP dependent protein kinase (cAMP kinase) was without effect. In cell-attached patches obtained from freshly isolated bovine tracheal smooth muscle cells, the open state probability (NPo) of maxi-KCa channels (conductance of approximately 260 picosiemens) was enhanced by 300 microM 8-(4-chlorophenylthio)-cGMP, a specific and potent activator of cGMP kinase. In contrast, 1 microM isoprenaline, 20 microM forskolin, and 3 mM 8-bromo-cAMP failed to enhance KCa channel activity. In excised inside-out patches, only the active fragment of cGMP kinase (but not that of cAMP kinase) increased NPo when applied to the cytosolic side of the patch. The enhancement of NPo by cGMP kinase was inhibited in CHO cells as well as in tracheal smooth muscle cells by the cGMP kinase inhibitor KT 5823 (1 microM) and the protein phosphatase (PP) inhibitors microcystin (5 microM) and okadaic acid (10 nM). The catalytic subunit of PP2A (but not that of PP1) mimicked the effect of cGMP kinase on NPo in excised inside-out patches. The results show that cGMP kinase regulates two different KCa channels in two unrelated cell types by the same indirect mechanism, which requires the activity of PP2A. The regulation of the KCa channel is specific for cGMP kinase and is not mimicked by cAMP kinase. PMID- 8702683 TI - Growth hormone promotes the association of transcription factor STAT5 with the growth hormone receptor. AB - Members of the cytokine/growth hormone (GH)/prolactin receptor superfamily transduce signals by association and activation of JAK tyrosine kinases. For GH receptor (GHR), both JAK2 and the GHR undergo tyrosine phosphorylation upon GH stimulation. Also, GH has recently been shown to activate the transcription factor STAT5 by tyrosine phosphorylation. In the present study, we demonstrate that GH induces rapid tyrosine phosphorylation of different isoforms of STAT5 in mouse L cells stably transfected with a cDNA encoding porcine GHR (pGHR). In this cell system, STAT5 directly interacts with the GHR in a GH-dependent manner. Additionally, GH-induced tyrosine phosphorylation of STAT5 and the interaction of STAT5 with GHR can be observed in mouse 3T3-F442A cells which express endogenous mouse GHR. Interestingly, when cDNAs encoding the two mouse STAT5 homologs (STAT5A and STAT5B) were individually transfected into mouse L cells expressing pGHR, only STAT5A demonstrated the ability to interact with the pGHR and subsequently underwent GH-dependent tyrosine phosphorylation. STAT5B did not. Therefore, the GH-dependent interaction of a particular STAT5 with tyrosine phosphorylated GHR may play an important role in GH-mediated signal transduction. PMID- 8702684 TI - Multiple interactions between hTAFII55 and other TFIID subunits. Requirements for the formation of stable ternary complexes between hTAFII55 and the TATA-binding protein. AB - We have cloned and characterized the human TATA-binding protein (TBP)-associated factor hTAFII55. hTAFII55, which has no known Drosophila counterpart, is present in both of the previously described TFIIDalpha and TFIIDbeta subpopulations. We describe the interactions of hTAFII55 with other subunits of the transcription factor TFIID. By cotransfection in COS cells, we show that hTAFII55 interacts with hTAFII250, hTAFII100, hTAFII28, hTAFII20, and hTAFII18, but not with hTAFII30 or TBP. Analysis of the binding of hTAFII55 and TBP to hTAFII28 deletion mutants indicates that distinct regions of hTAFII28 are required for these interactions. Although hTAFII55 does not interact by itself with TBP, stable ternary complexes containing hTAFII55 and TBP can be formed in the presence of hTAFII250, hTAFII100, or hTAFII28. These results not only show that hTAFII100 and hTAFII28 interact with TBP, but also that they can nucleate the formation of partial TFIID complexes. PMID- 8702685 TI - Cloning and characterization of a glucocorticoid-induced diacylglycerol kinase. AB - Diacylglycerol kinase (DGK) plays a key role in cellular processes by regulating the intracellular concentration of the second messenger diacylglycerol. We screened a hamster DDT1 smooth muscle cell library and isolated a unique, glucocorticoid-inducible cDNA with substantial homology to known DGKs. DGK activity was increased in lysates of insect cells infected with recombinant baculovirus containing this cDNA. Antibodies raised against expressed sequences recognized a glucocorticoid-inducible 130-140-kDa protein on immunoblots of DDT1 cell lysates. Thus, this sequence appears to be a new member of the DGK family that we refer to as DGKeta. Homology to other DGKs was apparent in domains that are thought to be important for DGK function including the cysteine-rich motifs and potential catalytic domains. DGKeta shares substantial homology with DGKdelta including the N-terminal pleckstrin homology domain. The tissue distribution of DGKeta message (determined by ribonuclease protection assays) and protein (determined by immunoblots) was broader than reported for other DGKs, indicating that DGKeta may play a more general role in regulating cellular DG levels than other DGKs. Heterogeneity among DGK family members indicates that individual DGKs may have unique functions. PMID- 8702686 TI - A cell wall-associated, receptor-like protein kinase. AB - Physical connections between higher plant cell walls and the plasma membrane have been identified visually, but the molecules involved in the contact are unknown. We describe here an Arabidopsis thaliana protein kinase, designated Wak1 for wall associated kinase, whose predicted extracytoplasmic domain contains several epidermal growth factor repeats and identity with a viral movement protein. Wak1 fractionates with insoluble material when plant tissue is ground in a variety of buffers and detergents, suggesting a tight association with the plant extracellular matrix. Immunocytochemistry confirms that Wak1 is associated with the cell wall. Enzymatic digestion of the cell wall allows the release of Wak1 from the insoluble cell wall fraction, and protease experiments indicate that Wak1 likely has a cytoplasmic kinase domain, and the EGF containing domain is extracellular. Wak1 is found in all vegetative tissues of Arabidopsis, and has relatives in other angiosperms, but not Chlamydomonas. We suggest that Wak1 is a good candidate for a physical continuum between the cell wall and the cytoplasm, and since the kinase is cytoplasmic, it also has the potential to mediate signals to the cytoplasm from the cell wall. PMID- 8702687 TI - Rac "insert region" is a novel effector region that is implicated in the activation of NADPH oxidase, but not PAK65. AB - The small GTPase Rac assembles with the cytosolic p47(phox) and p67(phox) and the membrane-associated flavocytochrome b558 to form the multicomponent respiratory burst oxidase. Mutation of amino acids in a region of Rac (residues 26-45), homologous to an effector region in Ras, was previously shown to interfere with Rac binding to the oxidase. Herein we have elucidated an additional region in Rac involved in regulating oxidase activity. Rho family small GTPases contain a 12 amino acid "insert" region (residues 124-135) that is not present in Ras. Point mutations in and deletion of this region were constructed and used for in vitro studies of the activation of PAK65 and NADPH oxidase. Apparent binding constants (based on EC50 values) of the mutant Rac proteins for the oxidase are at least 13 25-fold higher than for wild-type Rac. Mutations in the insert region versus the 26-45 effector region resulted in distinct kinetic consequences, pointing to different roles for these two protein regions: mutations in the insert region but not the 26-45 effector region resulted in an increase in the EC50 for p67(phox). Although mutations in the 26-45 amino acid effector region showed markedly diminished activation of both PAK and the NADPH oxidase, insert region mutations did not affect activation of PAK. We propose that the combinatorial use of the 26 45 effector region and the insert region provides the Rho family GTPases with versatility in their specificity for several downstream targets. PMID- 8702688 TI - Rabbit polymorphonuclear neutrophils form 35S-labeled S-sulfo-calgranulin C when incubated with inorganic [35S]sulfate. AB - Rabbit peritoneal polymorphonuclear neutrophils reduced inorganic [35S]sulfate to [35S]sulfite in vitro, concomitant with incorporation of 35S into a 10.68-kDa cytosolic protein as a S-[35S]sulfo-derivative. Amino-terminal sequencing of the purified protein identified calgranulin C, a member of the S100 protein family. cDNA clones of calgranulins B and C were isolated using oligonucleotide primers based on the established amino acid sequences of other mammalian calgranulins. The complete amino acid sequence of rabbit calgranulin C was deduced from the nucleotide sequence of the corresponding cDNA. It comprises 91 amino acid residues, has a calculated molecular mass of 10.52 kDa, has 74% identity with porcine calgranulin C, and shows high homology with other S100 calcium-binding proteins. Rabbit calgranulin C has a single cysteine residue at position 30, which we believe to be modified to S-[35S]sulfo-cysteine as a consequence of sulfate reduction by neutrophils. The formation of S-[35S]sulfo-calgranulin C appears to be a reaction specific to neutrophils. The specific radioactivity of calgranulin C from the neutrophil culture medium was 50-fold greater than that of the calgranulin C within the cells, suggesting that S-sulfation of calgranulin C might be associated with its secretion. PMID- 8702689 TI - Protein-tyrosine phosphatase 1B is a negative regulator of insulin- and insulin like growth factor-I-stimulated signaling. AB - To understand the physiological role of protein-tyrosine phosphatase 1B (PTPase 1B) in insulin and insulin-like growth factor-I (IGF-I) signaling, we established clonal cell lines overexpressing wild type or inactive mutant (C215S) PTPase 1B in cells overexpressing insulin (Hirc) or IGF-I (CIGFR) receptors. PTPase 1B overexpression in transfected cells was verified by immunoblot analysis with a monoclonal PTPase 1B antibody. Subfractionation of parental cells demonstrated that greater than 90% of PTPase activity was localized in the Triton X-100 soluble particulate (P1) cell fraction. PTPase activity in the P1 fraction of cells overexpressing wild type PTPase 1B was 3-6-fold higher than parental cells but was unaltered in all fractions from C215S PTPase 1B-containing cells. The overexpression of wild type and C215S PTPase 1B had no effects on intrinsic receptor kinase activity, growth rate, or general cell morphology. The effects of PTPase 1B overexpression on cellular protein tyrosine phosphorylation were examined by anti-phosphotyrosine immunoblot analysis. No differences were apparent under basal conditions, but hormone-stimulated receptor autophosphorylation and/or insulin receptor substrate tyrosine phosphorylation were inhibited in cells overexpressing wild type PTPase 1B and increased in cells expressing mutant PTPase 1B, in comparison with parental cells. Metabolic signaling, assessed by ligand-stimulated [14C]glucose incorporation into glycogen, was also inhibited in cells overexpressing active PTPase 1B and enhanced in cells containing C215S PTPase 1B. These data strongly suggest that PTPase 1B acts as a negative regulator of insulin and IGF-I signaling. PMID- 8702690 TI - Cyclic GMP induces oscillatory calcium signals in rat hepatocytes. AB - The ability of guanosine-3',5'-cyclic monophosphate (cGMP) to induce increases in the intracellular free calcium ion concentration ([Ca2+]i) was studied at the single cell level in fura-2-loaded rat hepatocytes. Both 8-bromo-cGMP (Br-cGMP) and dibutyryl cGMP (db-cGMP) produced oscillatory [Ca2+]i increases in hepatocytes. In addition, Br-cGMP increased the frequency of agonist-induced spiking or converted [Ca2+]i oscillations into sustained nonoscillatory [Ca2+]i responses. Addition of the nitric oxide donor sodium nitroprusside also produced oscillatory [Ca2+]i increases similar to those generated by cGMP analogues. In the absence of extracellular Ca2+, cGMP-induced [Ca2+]i responses were significantly reduced and mainly appeared as single transient [Ca2+]i increases. The effects of cGMP analogues do not appear to be mediated by a secondary increase in cAMP or activation of cAMP-dependent protein kinase (PKA), since [Ca2+]i responses to cGMP analogues were inhibited by the G-kinase inhibitor 8 bromoguanosine-3',5'-cyclic monophosphorothioate (Rp-Br-cGMP[S]). Both Br-cGMP and db-cGMP also increased [Ca2+]i in the presence of the PKA inhibitor 8 bromoadenosine-3',5'-cyclic monophosphorothioate (Rp-Br-cAMP[S]) and when the cGMP-inhibitable cAMP phosphodiesterase activity was inhibited by pretreatment with siguazodan. Br-cGMP stimulated the Mn2+-induced quench of compartmentalized fura-2 in intact hepatocytes, indicating a site of action at the level of the Ca2+ stores. This locus was further supported by the finding that pretreatment of hepatocytes with Br-cGMP potentiated submaximal inositol 1,4,5-trisphosphate (InsP3)-induced Mn2+ quench in subsequently permeabilized hepatocytes. db-cGMP also decreased PKA-mediated back phosphorylation of the hepatic type-1 InsP3 receptor, indicating that G-kinase phosphorylates the InsP3 receptor at sites targeted by PKA. These data indicate that phosphorylation of the hepatic InsP3 receptor by G-kinase increases the sensitivity to InsP3 for [Ca2+]i release and is associated with the production of [Ca2+]i oscillations in single rat hepatocytes. PMID- 8702691 TI - Phosphorylation of non-bleached rhodopsin in intact retinas and living frogs. AB - The photoresponse in retinal photoreceptors begins when a molecule of rhodopsin is excited by a photon of light. Photoexcited rhodopsin activates an enzymatic cascade including the G-protein transducin and cyclic GMP phosphodiesterase. As a result, cytoplasmic cyclic GMP concentration is decreased and the photoresponse is initiated. This process is terminated when rhodopsin is phosphorylated by rhodopsin kinase and subsequently blocked by a protein called arrestin. It has been noted by several investigators that light can cause phosphorylation of not only photoexcited but also non-excited rhodopsin in rod photoreceptors. A goal of this study was to determine how much non-bleached rhodopsin is phosphorylated. To determine how the structural integrity of the photoreceptor influences the extent of non-bleached rhodopsin phosphorylation, we studied the reaction in electropermeabilized rod outer segments, in rod outer segments still attached to isolated retinas and in living frogs. In the first two preparations, we found that the maximum extent of non-bleached rhodopsin phosphorylation was approximately 1% of the total rhodopsin pool. In living frogs, the maximal amount of non-bleached rhodopsin phosphorylation was approximately 2% of the total rhodopsin pool and occurred after prolonged illumination by the relatively dim light intensity of 20 lux. These data appear to exclude models for light adaptation that postulate high levels of phosphorylation of non-bleached rhodopsin in rod photoreceptors. PMID- 8702692 TI - A plant poly(A) polymerase requires a novel RNA-binding protein for activity. AB - We have purified a novel factor (PAP-III) that is a component of a multisubunit poly(A) polymerase from pea seedlings. This factor consists of one or more polypeptides with molecular masses of about 105 kDa and of a population of associated RNAs that can serve as substrates for polyadenylation. When these RNAs are separated from the 105-kDa polypeptides, polyadenylation becomes dependent upon exogenously added RNA. This RNA-dependent activity does not require the presence of a polyadenylation signal in the substrate, indicating that the activity under study is a nonspecific polyadenylation activity. One or more of the 105-kDa polypeptides could be cross-linked to the products of polyadenylation labeled with [alpha-32P]ATP and to exogenously added labeled RNAs. Cross-linking of the 105-kDa polypeptides to the products of polyadenylation was not affected by the presence of exogenously added competitors, whereas cross-linking to exogenous RNAs was diminished by excesses of RNA homopolymers. Exogenous RNAs could be polyadenylated by the combination of PAP-I + PAP-III, and this activity was diminished if the binding of the exogenous RNAs to PAP-III was prevented. We conclude from these studies that PAP-III is an RNA binding protein, that polyadenylation by the poly(A) polymerase occurs while the substrate RNAs are associated with this protein, and that the pea poly(A) polymerase can only polyadenylate those RNAs that are associated with PAP-III. PMID- 8702693 TI - Atrial chamber-specific expression of the slow myosin heavy chain 3 gene in the embryonic heart. AB - The quail slow myosin heavy chain 3 (slow MyHC 3) gene is expressed in the developing heart and in slow muscles of the developing limb. It is first expressed in the pulsatile cardiac tube in the embryo, and as the heart chamberizes its expression becomes restricted to the atria. To identify regulatory elements responsible for atrial-specific expression, the 5' upstream region of slow MyHC 3 gene was investigated. An atrial regulatory domain (ARD1) between -840 and -680 acts as an atrial cell-specific enhancer in primary cardiocyte cultures. ARD1 also specifies atrial-specific expression in vivo when the ARD1/heterologous promoter was introduced into developing chick embryos by a replication-competent retroviral vector. ARD1 is the first atrial cell-specific enhancer to be identified. Fine deletion and mutation analysis within ARD1 defined a 40-base pair vitamin D3 receptor-like element that controls atrial cell specific expression of the slow MyHC 3 gene by inhibiting its expression in ventricular cardiocytes. PMID- 8702694 TI - Functional diversity of alternatively spliced isoforms of Drosophila Ca2+/calmodulin-dependent protein kinase II. A role for the variable domain in activation. AB - Isoforms of calcium/calmodulin-dependent protein kinase II from Drosophila (R1-R6 and R3A) showed differential activation by two series of mutant calmodulins, B1K B4K and B1Q-B4Q. These mutant calmodulins were generated by changing a glutamic acid in each of the four calcium binding sites to either glutamine or lysine, altering their calcium binding properties. All mutations produced activation defects, with the binding site 4 and B1Q mutants the most severe. Activation differed substantially between isoforms. R4, R5, and R6 were the least sensitive to mutations in calmodulin, while R1, R3, and R3A were the most sensitive. Activation of R1 and R2 by B4K and activation of R3 and R3A by B2K and B2Q produced significant (6-fold and almost 3-fold, respectively) differences in Kact between isoforms that differ structurally by a single amino acid. These differences could not be accounted for by differential binding, as all isoforms showed almost identical binding patterns with the mutants. High binding affinity did not always correlate with ability to increase enzyme activity, implying that activation occurs in at least two steps. The isoform-specific differences seen in this study reflect a role for the COOH-terminal variable region in activation of CaM kinase II. PMID- 8702695 TI - Inhibition of initiation of simian virus 40 DNA replication in infected BSC-1 cells by the DNA alkylating drug adozelesin. AB - Adozelesin is a member of a family of extraordinarily cytotoxic DNA damaging agents that bind to the DNA minor groove in a sequence-specific manner and form covalent adducts with adenines. Previous studies employing purified enzymes and adozelesin-modified template DNAs suggested that adozelesin-DNA adducts inhibit DNA replication at the level of nascent DNA chain elongation. In this study, neutral/neutral two-dimensional agarose gel electrophoresis was employed to analyze simian virus 40 (SV40) DNA replication intermediates recovered from adozelesin-treated SV40 virus-infected cells. SV40 replication intermediates rapidly disappeared from infected cells when they were treated with adozelesin, but not when the cells were also treated with aphidicolin to block maturation of replicating SV40 DNA. We conclude that the disappearance of SV40 replication intermediates induced by adozelesin treatment was a consequence of maturation of these intermediates in the absence of new initiation events. Adozelesin inhibition of nascent chain elongation is first observed at concentrations above those needed to block initiation. Adozelesin treatment inhibits SV40 DNA replication at concentrations that produce adducts on just a small fraction of the intracellular population of SV40 DNA molecules. PMID- 8702696 TI - The K65R mutation confers increased DNA polymerase processivity to HIV-1 reverse transcriptase. AB - The K65R mutation in HIV-1 reverse transcriptase (RT) is associated with viral cross-resistance to 2',3'-dideoxyinosine, 2',3'-dideoxycytidine, and 2',3' dideoxy-3'-thiacytidine. We have found that in vitro DNA synthesis by K65R RT is significantly more processive than that of wild type (wt) RT. Depending on the template/primer (T/P) used, the total incorporation of nucleotides under single processive cycle conditions was 20-50% higher with K65R RT than with wt RT. With heteropolymeric T/P, the total incorporation of dNMP by K65R and wt RT was similar under continuous DNA synthesis reaction conditions. However, under single processive cycle conditions, the rate of full-length polymerization product synthesis by K65R RT was about 2-fold higher than that by wt RT. We also found a decreased rate of T/P dissociation during K65R RT DNA synthesis, which is consistent with the increased processivity of the enzyme. We postulate that the increased processivity of the K65R RT may be a compensatory response to the decreased affinity of this mutant for certain dNTP substrates, allowing normal viral replication kinetics. PMID- 8702697 TI - Globin and globin gene structure of the nerve myoglobin of Aphrodite aculeata. AB - The globin of the nerve cord of the polychaete annelid Aphrodite aculeata was isolated and purified to homogeneity. The native molecule has a pI of 6.3 and acts as a dimer of two identical Mr 15, 644.5 polypeptide chains as determined by electrospray mass spectrometry. It has an average affinity for oxygen (P50 = 1.24 torr) resulting from fast association (kon = 170 X 10(6) M-1 . s-1) and dissociation rates (koff = 360 s-1). The partial primary structure of this nerve globin was determined at the protein level and completed and confirmed by translation of the cDNA sequence. The globin chain has 150 amino acid residues and a calculated Mr of 15, 602.69 strongly suggesting that the amino terminus is acetylated. The absence of a leader sequence and the lack of Cys at the positions NA2 and H9 needed for the formation of the high Mr complexes found in extracellular annelid globins classify the Aphrodite globin with the cellular globin species. The Aphrodite nerve globin is unlikely to represent a separate globin family, as cDNA derived primers detect globin messenger RNA in muscle, gut, and pharynx tissue as well. The gene encoding this globin species is interrupted by a single intron, inserted at position G7.0. Comparison to other globin gene structures strongly suggest that introns can be lost independently, rather than simultaneously as a result of a single conversion event as suggested previously (Lewin, R. (1984) Science 226, 328). PMID- 8702698 TI - Turnover and translation of in vitro synthesized messenger RNAs in transfected, normal cells. AB - We have developed a novel system to examine intracellular mRNA decay pathways in the absence of transcriptional blockade. In vitro transcribed, capped, and adenylated granulocyte-macrophage colony stimulating factor (GM-CSF) or globin mRNAs were introduced by particle-mediated gene transfer into primary cultures of normal peripheral blood mononuclear cells. Transfected wild-type, human GM-CSF (hGM-AUUUA) mRNA decayed rapidly (t1/2 = 9 min), while a mutated version lacking AUUUA repeats (hGM-AUGUA) was significantly more stable (t1/2 = 30 min). A truncated GM-CSF mRNA lacking the entire 3'-UTR (hGM-Delta3'-UTR) was still more stable (t1/2 = 80 min) demonstrating the existence of non-AUUUA, 3'-UTR destabilizing domains. Transfected beta-globin mRNA was very stable, decaying with a half-life of >360 min. Transfected mRNAs were >90% polysome associated with transgenic protein detectable within 15 min of transfection. The most stable GM-CSF mRNAs were not associated with maximal GM-CSF protein production. Agents known or hypothesized to interfere with mRNA decay, including cycloheximide, phorbol ester, or actinomycin D, stabilized both hGM-AUUUA and hGM-AUGUA mRNAs. These data demonstrate the presence of 3'-UTR, destabilizing, and translational regulatory elements outside of the AUUUA repeats and unambiguously show that actinomycin D at concentrations commonly used to inhibit transcription stabilizes cytokine mRNAs. PMID- 8702699 TI - Calcineurin mediates calcium-induced potentiation of adenylyl cyclase activity in dispersed chief cells from guinea pig stomach. Further evidence for cross-talk between signal transduction pathways that regulate pepsinogen secretion. AB - In cholera toxin-treated gastric chief cells, incubation with a cholinergic agonist (carbamylcholine), a regulatory peptide (cholecystokinin), or a calcium ionophore (A23187) causes a dose- and time-dependent potentiation of cAMP levels. Because this augmented response is calcium/calmodulin-dependent, we hypothesized that it was mediated by calcineurin (protein phosphatase 2B). To test this hypothesis, we examined the actions of calcineurin inhibitors on secretagogue induced potentiation of cAMP levels in guinea pig chief cells. Preincubation of cells with 0.1 microM FK-506 completely prevented carbachol-induced augmentation of cAMP levels and pepsinogen secretion from cholera toxin-treated cells. Cyclosporin-A, another calcineurin inhibitor, also prevented the augmented cAMP response. FK-506 and cyclosporin inhibited augmentation of cAMP levels following treatment with cholecystokinin(26-33) and A23187, but not the smaller increase in cAMP following treatment with a phorbol ester that activates protein kinase C. Hence, the actions of calcineurin inhibitors were limited to secretagogues that increase cellular calcium. Rapamycin, an agent that competes with FK-506 for the immunophilin, FK binding protein 12, does not inhibit calcineurin. In the present study, preincubation with rapamycin did not prevent carbachol-induced augmentation of cAMP levels in cholera toxin-treated chief cells. However, a molar excess of rapamycin reversed the inhibitory actions of FK-506. These experiments provide further evidence that the actions of FK-506 on cholera toxin treated gastric chief cells are caused by its inhibitory actions on calcineurin. FK-506 also inhibited potentiation of cAMP levels when carbachol was added to cells that were preincubated with forskolin, an agent that directly activates adenylyl cyclase. We conclude that, in gastric chief cells, calcineurin mediates cross-talk between the calcium/calmodulin and adenylyl cyclase signaling pathways. PMID- 8702700 TI - The putative substrate recognition loop of Escherichia coli ribonuclease H is not essential for activity. AB - The RNase H family of enzymes catalyzes the hydrolysis of RNA from RNA DNA hybrids in a divalent metal-dependent fashion. To date, structure/function studies have focused on two members of this family: Escherichia coli RNase HI, a small monomeric protein; and human immunodeficiency virus, type I (HIV) RNase H, a domain of HIV reverse transcriptase. The isolated RNase H domain from HIV reverse transcriptase can be expressed independently and shares significant structural homology with its E. coli homologue; however, unlike the bacterial protein, it is inactive. The most notable difference between the inactive domain from HIV and the active E. coli protein is a basic helix/loop sequence, present in E. coli but absent from the HIV homologue. Substitution of this basic region into the HIV domain partially restores its activity and increases its thermodynamic stability. By deleting the basic helix/loop region, we have modeled the structural difference between these two polypeptides onto the E. coli homologue. Surprisingly, the resulting mutant protein is active in Mn2+-dependent fashion. Therefore, the basic helix/loop is not required for RNase H activity. PMID- 8702701 TI - Converting parathyroid hormone-related peptide (PTHrP) into a potent PTH-2 receptor agonist. AB - Most of the bone and kidney-related functions of parathyroid hormone (PTH) and parathyroid hormone-related peptide (PTHrP) are thought to be mediated by the PTH/PTHrP receptor. Recently, a homologous receptor, the PTH-2 receptor, was obtained from rat and human brain cDNA libraries. This receptor displayed the remarkable property of responding potently to PTH, but not to PTHrP. To begin to define residues involved in the ligand specificity of the PTH-2 receptor, we studied the interaction of several PTH/PTHrP hybrid ligands and other related peptide analogs with the human PTH-2 receptor. The results showed that two sites in PTH and PTHrP fully account for the different potencies that the two ligands exhibited with PTH-2 receptors; residue 5 (His in PTHrP and Ile in PTH) determined signaling capability, while residue 23 (Phe in PTHrP and Trp in PTH) determined binding affinity. By changing these two residues of PTHrP to the corresponding residues of PTH, we were able to convert PTHrP into a ligand that avidly bound to the PTH-2 receptor and fully and potently stimulated cAMP formation. Changing residue 23 alone yielded [Trp23]hPTHrP-(1-36), which was an antagonist for the PTH-2 receptor, but a full agonist for the PTH/PTHrP receptor. Residues 5 and 23 in PTH and PTHrP thus play key roles in signaling and binding interactions, respectively, with the PTH-2 receptor. Receptor-selective agonists and antagonists derived from these studies could help to identify the biological role of the PTH-2 receptor and to map specific sites of ligand-receptor interaction. PMID- 8702702 TI - Overexpression of the normal phosphoribosylpyrophosphate synthetase 1 isoform underlies catalytic superactivity of human phosphoribosylpyrophosphate synthetase. AB - To define the enzymatic and genetic basis of X-linked phosphoribosylpyrophosphate synthetase (PRS) catalytic superactivity, we measured concentrations of X-linked PRS1 and PRS2 isoforms in cultured fibroblasts and lymphoblasts by immunoblotting after separation by polyacrylamide-urea isoelectric focusing. PRS1 comprised >80% of measurable PRS isoforms in all fibroblast strains, but PRS1 concentrations in cells from six affected males exceeded those in normal cells by 2-6-fold. PRS absolute specific activities (activity per mg of PRS isoforms) were comparable in all fibroblast strains and in purified recombinant normal PRS1, confirming selectively increased levels of PRS1 isoform as the enzymatic basis of PRS catalytic superactivity. Cloning, sequencing, and expression of normal subject- and patient-derived PRS cDNAs predicted normal translated region sequences for both PRS isoforms and revealed no differences in catalytic properties of recombinant PRS1. Normal and patient PRPS1 transcribed but untranslated DNA sequences were also identical. Northern blot analysis showed selective increase in relative concentrations of PRS1 transcripts in patient fibroblasts. In PRS catalytic superactivity, overexpression of the normal PRS1 isoform thus appears to result from an altered pretranslational mechanism of PRPS1 expression. In lymphoblasts, however, expression of this alteration is attenuated, explaining the absence of phenotypic expression of PRS catalytic superactivity in these cells. PMID- 8702703 TI - Activation of the human androgen receptor through a protein kinase A signaling pathway. AB - Aberrant activation of the androgen receptor through signaling pathways independent of androgen may be responsible for the progression of prostate tumors to the rapidly proliferating androgen-independent state. In this study, the effects of protein kinase A modulators on human androgen receptor activity were tested. Using an adenoviral DNA delivery system, we demonstrate that the androgen receptor can be activated by a protein kinase A activator, forskolin, in the absence of androgen when androgen receptor is co-transfected into monkey kidney CV1 cells or human prostate PC-3 cells with androgen-responsive reporters. Immunoblotting reveals that there is no significant change in androgen receptor protein level following forskolin treatment, suggesting that the enhanced activity is due to activation of the receptor. This activation can be blocked by a protein kinase A inhibitor peptide. Two potent anti-androgens, casodex and flutamide, can significantly reduce this activation, confirming that the ligand independent pathway is an androgen receptor-mediated phenomenon. An intact DNA binding domain of the receptor is critical for this alternate signaling pathway since mutants with reduced DNA binding ability are inactive. The phosphorylation status of the androgen receptor or associated proteins may critically modulate receptor activity and should be considered when designing improved approaches to prostate cancer therapy. PMID- 8702704 TI - In vivo analyses of interactions between SecE and SecY, core components of the Escherichia coli protein translocation machinery. AB - We have carried out structure-function studies on the cytoplasmic membrane protein, SecE, a component of the Escherichia coli secretion machinery. SecE, along with SecY, form a complex in the cytoplasmic membrane essential for protein translocation. By directed mutagenesis, we altered highly conserved residues of the second cytoplasmic domain (CD2) and of the COOH-terminal periplasmic region (PD2) of SecE. These mutants, as well as previously constructed mutations in the third membrane-spanning segment of SecE (MSS3), were tested for their ability to complement a secE null mutation, for their effects on protein export in vivo, and for their ability to form a stable complex with SecY. Most single mutations at the conserved positions in CD2 caused secretion defects, but had little effect on growth at 37 degrees C. Double mutations in CD2, or the introduction or removal of proline residues, affected growth and protein translocation more severely. Co immunoprecipitations of SecE and SecY revealed that all mutant proteins, except those altered in PD2, destabilized the SecE-SecY complex. These results suggest that several regions contribute to the formation of a stable SecE-SecY complex but the elimination of a single contact point does not necessarily affect the functionality of the complex. PMID- 8702705 TI - 19F NMR studies with 2'-F-2'-deoxyarabinoflavoproteins. AB - Apoproteins of several flavoproteins were reconstituted with 2'-F-2' deoxyarabinoflavins and studied by 19F NMR and absorption spectroscopy. Extensive protein-fluorine interactions were observed by large chemical shift changes on binding to the apoprotein of Old Yellow Enzyme (apoOYE) and apoflavodoxin, whereas binding to apoglucose oxidase and apo -amino acid oxidase (apoDAAO) resulted in minimal interactions. Modification at the flavin 2'-position in OYE resulted in a substantial decrease in the binding affinity of the flavin, possibly from the disruption of two important hydrogen bonds to Pro-35 and Arg 243. 19F NMR studies of complexes of OYE with testosterone, cyclohexenone, and beta-estradiol suggest that phenols and alpha,beta-unsaturated ketones orient differently at the active site on binding. The two separate one-electron potentials for the EFlox/EFlsq and EFlsq/EFlred couples were different for the reconstituted OYE. With native enzyme, there is 15-20% thermodynamic stabilization of the anionic flavin semiquinone, while no detectable amount of semiquinone was observed with modified OYE. This change in potential was further substantiated by blue shifts for the maxima of the modified protein-phenol charge transfer complexes. In accordance with the crystal structure of the OYE-p-OH benzaldehyde complex (Fox, K.M. & Karplus, P.A. (1994) Structure 2, 1089-1105), 19F NMR studies with the modified OYE-2,4-F2-phenol suggest strong interaction between the para-fluorine of the phenol and Tyr-375. PMID- 8702706 TI - Delineation of transmembrane domains of the Na+/H+ exchanger that confer sensitivity to pharmacological antagonists. AB - Plasma membrane Na+/H+ exchanger (NHE) isoforms NHE1 and NHE3 exhibit very different sensitivities to amiloride and its 5-amino-substituted analogues, benzoyl guanidinium derivatives (e.g. (3-methylsulfonyl-4 piperidinobenzoyl)guanidine methanesulfonate (HOE694)), and cimetidine. To define structural domains that confer differential sensitivity to these antagonists, unique restriction endonuclease sites were engineered into cDNAs for each isoform near the regions that encode the putative membrane-spanning domains. These new sites did not modify their pharmacological properties and allowed several chimeric Na+/H+ exchangers to be constructed by exchanging homologous segments. The modified parental (E1' and E3') and chimeric molecules were stably expressed in exchanger-deficient Chinese hamster ovary AP-1 cells and assayed for their sensitivities to amiloride, ethylisopropylamiloride, HOE694, and cimetidine. Most chimeras showed drug sensitivities corresponding to the dominant parental segment. However, interchanging a 66-amino acid segment containing the putative ninth transmembrane (M9) domain and its adjacent loops caused reciprocal alterations in the sensitivities of E1' and E3' to all antagonists. In addition, substituting the first five putative membrane-spanning domains of E3' with the corresponding region of E1' modestly reduced the transporter's sensitivity to cimetidine but not the other compounds. These data indicate that the protein segment between M8 and M10 may be a major site of interaction with these antagonists, although other regions modestly influence sensitivity to certain drugs. PMID- 8702707 TI - Nucleoside diphosphate kinase. Investigation of the intersubunit contacts by site directed mutagenesis and crystallography. AB - NDP kinase from Dictyostelium was mutated by site-directed mutagenesis at positions indicated by structural data to be involved in the trimer interface. The mutants were substitutions at residue Pro-100 (P100S and P100G) and deletions of 1-5 residues at the C terminus. Single mutants yielded proteins that kept both activity and hexameric structure. However, they were severely affected in their stability toward temperature and urea denaturation. When the P100S mutation was combined with any of the C-terminal deletions, the enzyme lost most of its activity and dissociated into dimers. Crystallographic analysis of the P100S protein was performed at 2.6 A resolution. The x-ray structure showed no direct alteration of intersubunits contacts at residue 100, but an induced disruption of the interaction between Asp-115 and the C terminus of another subunit. The substitution of proline 100 to serine corresponds to the Killer-of-prune mutation in Drosophila. Consequences of the mutation are discussed in view of the structural and biochemical properties observed in the mutant Dictyostelium protein. PMID- 8702708 TI - Anatomy of TRAF2. Distinct domains for nuclear factor-kappaB activation and association with tumor necrosis factor signaling proteins. AB - The tumor necrosis factor (TNF) receptor-associated factor (TRAF) family of proteins interact with and transduce signals for members of the TNF receptor superfamily. TRAF1, TRAF2, and TRAF3 share a conserved C-terminal TRAF domain. TRAF2 plays a key role in transducing signals for activation of the transcription factor nuclear factor-kappaB (NF-kappaB). We have performed extensive mutational analysis on TRAF2, examining the requirements for NF-kappaB activation, self association, and interaction with other molecules involved in TNF signaling. Examination of point mutants and TRAF2-TRAF3 chimeric proteins indicates that the N-terminal RING finger and two adjacent zinc fingers of TRAF2 are required for NF kappaB activation. The two distinct TRAF-N and TRAF-C subdomains of the TRAF domain appear to independently mediate self-association and interaction with TRAF1. Interaction of TRAF2 with TNF-R2 and TRADD requires sequences at the C terminus of the TRAF-C domain, whereas interaction with the protein kinase receptor-interacting protein V(RIP) occurs via sequences at the N terminus of the TRAF-C domain. Thus, distinct domains of TRAF2 are involved in recruitment and signaling functions. PMID- 8702709 TI - Mammary derived growth inhibitor is not a distinct protein but a mix of heart type and adipocyte-type fatty acid-binding protein. AB - The amino acid sequence of the mammary derived growth inhibitor (MDGI) from bovine mammary gland (Bohmer, F.-D., Kraft, R., Otto, A. , Wernstedt, C., Hellman, U., Kurtz, A., Muller, T., Rohde, K., Etzold, G., Lehmann, W., Langen, P., Heldin, C.-H., and Grosse, R. (1987) J. Biol. Chem. 262, 15137-15143) revealed 95% identity to bovine heart fatty acid-binding protein (H-FABP), explaining the observed immunocross-reactivity. However, a cDNA encoding MDGI has not been found to date. Artificial MDGI cDNA was expressed in an in vitro transcription/translation assay. Analysis by isoelectric focusing of the immunoprecipitated in vitro translation products of lactating bovine mammary gland mRNA did not indicate a protein corresponding to the in vitro translation product of artificial MDGI mRNA. Moreover, two-dimensional electrophoresis of bovine mammary gland proteins confirmed the absence of a protein with the pI of the in vitro translated artificial MDGI mRNA in bovine mammary gland and instead revealed, apart from H-FABP, an unknown protein that was recognized by anti-H FABP antibodies. From lactating bovine mammary gland the cDNA for adipocyte fatty acid-binding protein (A-FABP) was cloned. The in vitro translation of recombinant mRNA derived from this cDNA yielded a polypeptide that behaved like the unknown immunoreactive protein. Western blotting and immunofluorescence using monospecific antibodies demonstrated the coexistence of H-FABP and A-FABP in the lactating mammary gland. Taking into account that deviations of the MDGI sequence from the bovine H-FABP sequence correspond with A-FABP we attribute the structure originally reported as MDGI to a mix of these proteins. PMID- 8702710 TI - Human phagocytes employ the myeloperoxidase-hydrogen peroxide system to synthesize dityrosine, trityrosine, pulcherosine, and isodityrosine by a tyrosyl radical-dependent pathway. AB - Myeloperoxidase, a heme protein secreted by activated phagocytes, may be a catalyst for lipoprotein oxidation in vivo. Active myeloperoxidase is a component of human atherosclerotic lesions, and atherosclerotic tissue exhibits selective enrichment of protein dityrosine cross-links, a well characterized product of myeloperoxidase. Tyrosylation of lipoproteins with peroxidase-generated tyrosyl radical generates multiple protein-bound tyrosine oxidation products in addition to dityrosine. The structural characterization of these products would thus serve as an important step in determining the role of myeloperoxidase in lipoprotein oxidation in the artery wall. We now report the identification and characterization of four distinct tyrosyl radical addition products generated by human phagocytes. Activated neutrophils synthesized three major fluorescent products from -tyrosine; on reverse phase HPLC, each compound coeluted with fluorescent oxidation products formed by myeloperoxidase. We purified the oxidation products to apparent homogeneity by cation and anion exchange chromatographies and identified the compounds as dityrosine (3,3'-dityrosine), trityrosine (3,3',5',3"-trityrosine) and pulcherosine (5-[4"-(2-carboxy-2 aminoethyl)phenoxy]3, 3'-dityrosine) by high resolution NMR spectroscopy and mass spectrometry. Additionally, we have found that dityrosine is a precursor to trityrosine, but not pulcherosine. In a search for a precursor to pulcherosine, we identified isodityrosine (3-[4'-(2-carboxy-2-aminoethyl)phenoxy]tyrosine), a non-fluorescent product of L-tyrosine oxidation by human phagocytes. Our results represent the first identification of this family of tyrosyl radical addition products in a mammalian system. Moreover, these compounds may serve as markers specific for tyrosyl radical-mediated oxidative damage in atherosclerosis and other inflammatory conditions. PMID- 8702711 TI - Heterologous desensitization of the glucagon-like peptide-1 receptor by phorbol esters requires phosphorylation of the cytoplasmic tail at four different sites. AB - Glucagon-like peptide-1 stimulates glucose-induced insulin secretion by binding to a specific G protein-coupled receptor that activates the adenylyl cyclase pathway. We previously demonstrated that heterologous desensitization of the receptor by protein kinase C correlated with phosphorylation in a 33-amino acid long segment of the receptor carboxyl-terminal cytoplasmic tail. Here, we determined that the in vivo sites of phosphorylation are four serine doublets present at positions 431/432, 441/442, 444/445, and 451/452. In vitro phosphorylation of fusion proteins containing mutant receptor C-tails, however, indicated that whereas serines at position 431/432 were good substrates for protein kinase C (PKC), serines 444/445 and 451/452 were poor substrates, and serines 441/442 were not substrates. In addition, serine 416 was phosphorylated on fusion protein but not in intact cells. This indicated that in vivo a different PKC isoform or a PKC-activated kinase may phosphorylate the receptor. The role of phosphorylation on receptor desensitization was assessed using receptor mutants expressed in COS cells or Chinese hamster lung fibroblasts. Mutation of any single serine doublet to alanines reduced the extent of phorbol 12-myristate 13-acetate-induced desensitization, whereas substitution of any combination of two serine doublets suppressed it. Our data thus show that the glucagon-like peptide-1 receptor can be phosphorylated in response to phorbol 12 myristate 13-acetate on four different sites within the cytoplasmic tail. Furthermore, phosphorylation of at least three sites was required for desensitization, although maximal desensitization was only achieved when all four sites were phosphorylated. PMID- 8702712 TI - An interface of interaction between photoreceptor cGMP phosphodiesterase catalytic subunits and inhibitory gamma subunits. AB - Cyclic guanosine 5'-monophosphate (cGMP) phosphodiesterase (PDE) regulates the level of cGMP on transduction of a visual signal in vertebrate photoreceptor cells. Two identical inhibitory PDE gamma subunits (Pgammas) block catalytic activity of PDE-alpha and -beta subunits (Palphabeta) in the dark. The primary regions of Pgamma involved in the interaction with Palphabeta are a central polycationic region, Pgamma-24-45, and a C-terminal region of Pgamma. Recently, we have shown that the C-terminal region of Pgamma, which is the major Pgamma inhibitory domain, blocks PDE activity by binding to the catalytic site of PDE (Artemyev, N. O., Natochin, M., Busman, M., Schey, K. L., and Hamm, H. E. (1996) Proc. Natl. Acad. Sci. U. S. A. 93, 5407-5412). Here, we localize the site on the rod cGMP PDE alpha subunit that binds to the central polycationic domain of Pgamma. This site is located within a region that links a second noncatalytic cGMP binding site with the catalytic domain of PDE. A polypeptide coresponding to this region, Palpha-461-553, expressed as a glutathione S-transferase fusion protein in Escherichia coli and isolated after cleavage of the fusion protein with thrombin, blocks inhibition of PDE activity by Pgamma. In addition, Palpha 461-553 binds to the Pgamma-24-45 region (Kd, 7 microM), as measured by a fluorescent increase in a Pgamma-24-45Cys peptide labeled with 3-(bromoacetyl)-7 diethylaminocoumarin. The Palpha-461-553 region was further characterized by using a set of synthetic peptides. A peptide corresponding to residues 517-541 of Palpha (Palpha-517-541) effectively suppressed inhibition of PDE activity by Pgamma and bound to Pgamma-24-45Cys labeled with 3-(bromoacetyl)-7 diethylaminocoumarin (Kd, 22 microM). Palpha-517-541 also competes with the activated rod G-protein alpha-subunit for binding to Pgamma labeled with lucifer yellow vinyl sulfone. This suggests that light activation of rod PDE by the G protein transducin involves competition between transducin alpha-guanosine 5' triphosphate and Palpha-517-541 for binding to the Pgamma-24-45 region. Based on the results, we propose a linear model of interactions between catalytic and inhibitory PDE subunits. PMID- 8702713 TI - Identification of thromboxane A2 synthase active site residues by molecular modeling-guided site-directed mutagenesis. AB - Human thromboxane A2 synthase (TXAS) exhibits spectral characteristics of cytochrome P450 but lacks monooxygenase activity. Its distinctive amino acid sequence makes TXAS the sole member of family 5 in the P450 superfamily. To better understand the structure-function relationship of this unusual P450, we have recently constructed a three-dimensional model for TXAS using P450BM-3 as the template (Ruan, K.-H., Milfeld, K., Kulmacz, R. J., and Wu, K. K. (1994) Protein Eng. 7, 1345-1551) and have identified a potential active site region. The catalytic roles of several putative active site residues were evaluated using selectively mutated recombinant TXAS expressed in COS-1 cells. Mutation of Ala 408 to Glu or Arg-413 to Gly led to a complete loss of enzyme activity despite expression of mutant protein levels equivalent to that of the wild-type TXAS. Mutation of Ala-408 to Gly or Leu retained the enzyme activity at levels of 30 or 40%, respectively. This suggests that Ala-408 provides a hydrophobic environment for substrate binding. Mutation of Arg-413 to Lys or Gln completely abolished the enzyme activity, indicating that this residue is essential to catalytic activity and supports its identification as an active site residue. Mutation of Arg-410 to Gly or Glu-433 to Ala resulted in >50% reduction in the enzyme activity without appreciably altering mutant protein expression, consistent with a more subtle effect of these residues on TXAS catalytic efficiency. Mutation of residues predicted to be involved in binding the heme prosthetic group, including the heme thiolate ligand Cys-480, Arg-478, Phe-127, and Asn-110, each markedly reduced the expressed protein level and abolished enzyme activity. This suggests that proper heme binding is important to synthesis or stability of recombinant TXAS. Mutation of Ile-346, which corresponds to P450cam-Thr-252, an essential amino acid involved in dioxygen bond scission, to Thr increased the enzymatic activity by 40%, suggesting that oxygen bond cleavage is not a rate-limiting step in thromboxane A2 biosynthesis. The present results from site-directed mutagenesis support the overall structure of the TXAS active site predicted by homology modeling and have allowed refinement of the position of bound substrate. PMID- 8702714 TI - Differences between two tight ADP binding sites of the chloroplast coupling factor 1 and their effects on ATPase activity. AB - Purified chloroplast ATP synthase (CF1) contains 1.2-2 mol of tightly bound ADP/mol of enzyme that resists removal by gel filtration or dialysis. CF1 was depleted of its endogenous nucleotide by treatment with alkaline phosphatase. Tightly bound nucleotide was demonstrated not to have an essential structural role. CF1 depleted of endogenous nucleotide retains its ability to catalyze Ca2+- and Mg2+-dependent ATPase activity and is not more sensitive to cold inactivation than untreated CF1. 2'(3')-O-Trinitrophenyladenosine 5'-diphosphate (TNP-ADP) binds tightly to two sites on nucleotide-depleted CF1, binding to either site at a faster rate than that of exchange of bound nucleotide for medium nucleotide. The nucleotide-depleted enzyme binds about one additional mol of TNP-ADP/mol of CF1, indicating that there is a tight TNP-ADP binding site that does not exchange readily with medium nucleotide. It is MgADP in this nonexchanging site, not the easily exchanging ADP binding site, that is responsible for the MgADP-induced inhibition of the ATPase activity. The rate of exchange of tightly bound ADP from CF1 matches the rate at which the Mg2+ATPase activity of CF1 is activated but is not itself responsible for the activation. PMID- 8702715 TI - Identification of the ATP binding domain of recombinant human 40-kDa 2',5' oligoadenylate synthetase by photoaffinity labeling with 8-azido-[alpha-32P]ATP. AB - Three isoforms of the interferon-inducible 2',5'-oligoadenylate (2-5A) synthetase that require double-stranded RNA have been isolated and cloned. However, identification of the amino acid(s) of 2-5A synthetase directly interacting with ATP is crucial to the elucidation of the mechanism of the enzymatic conversion of ATP to 2',5'-oligoadenylates by 2-5A synthetase. Recombinant human 40-kDa 2-5A synthetase has been expressed as a glutathione S-transferase fusion protein in E. coli and purified to near homogeneity in milligram quantities. The azido photoprobe, 8-azido-[alpha-32P]ATP, has been used to identify the ATP binding domain of the recombinant human 40-kDa 2-5A synthetase. Specific covalent photoincorporation of 8-azido-[alpha-32P]ATP into the 2-5A synthetase, tryptic digestion of the covalently 32P-labeled enzyme, isolation of the photolabeled phosphopeptide by metal (Al3+) chelate chromatography, and high pressure liquid chromatography identified a 32P-pentapeptide, which has been assigned to the ATP binding domain of 2-5A synthetase. The radioactive pentapeptide has the sequence D196FLKQ200 in which the photoprobe, 8-azido-[alpha-32P]ATP, chemically modified the amino acid lysine 199. The catalytic importance of Lys199 was further established by mutation of lysine 199 to arginine 199 and histidine 199 using site-directed mutagenesis. The K199R and K199H recombinant human 40-kDa 2-5A synthetase mutants bind 8-azido-ATP and the allosteric activator, poly(I) poly(C) but are enzymatically inactive. These photoaffinity labeling and mutation data strongly suggest that lysine 199 is essential for the formation of a productive 2 5A synthetase-ATP-double-stranded RNA complex for the enzymatic conversion of ATP to 2-5A. PMID- 8702716 TI - Probing the structure and function of the tachykinin neurokinin-2 receptor through biosynthetic incorporation of fluorescent amino acids at specific sites. AB - A general method for understanding the mechanisms of ligand recognition and activation of G protein-coupled receptors has been developed. A study of ligand receptor interactions in the prototypic seven-transmembrane neurokinin-2 receptor (NK2) using this fluorescence-based approach is presented. A fluorescent unnatural amino acid was introduced at known sites into NK2 by suppression of UAG nonsense codons with the aid of a chemically misacylated synthetic tRNA specifically designed for the incorporation of unnatural amino acids during heterologous expression in Xenopus oocytes. Fluorescence-labeled NK2 mutants containing an unique 3-N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)-2,3-diaminopropionic acid (NBD-Dap) residue at either site 103, in the first extracellular loop, or 248, in the third cytoplasmic loop, were functionally active. The fluorescent NK2 mutants were investigated by microspectrofluorimetry in a native membrane environment. Intermolecular distances were determined by measuring the fluorescence resonance energy transfer (FRET) between the fluorescent unnatural amino acid and a fluorescently labeled NK2 heptapeptide antagonist. These distances, calculated by the theory of Forster, permit to fix the ligand in space and define the structure of the receptor in a molecular model for NK2 ligand receptor interactions. Our data are the first report of the incorporation of a fluorescent unnatural amino acid into a membrane protein in intact cells by the method of nonsense codon suppression, as well as the first measurement of experimental distances between a G protein-coupled receptor and its ligand by FRET. The method presented here can be generally applied to the analysis of spatial relationships in integral membrane proteins such as receptors or channels. PMID- 8702717 TI - Amino acid residues critical for the interaction between bacteriophage T7 DNA polymerase and Escherichia coli thioredoxin. AB - Upon infection of Escherichia coli, bacteriophage T7 annexes a host protein, thioredoxin, to serve as a processivity factor for its DNA polymerase, T7 gene 5 protein. In a previous communication (Himawan, J., and Richardson, C. C. (1992) Proc. Natl. Acad. Sci. U. S. A. 89, 9774-9778), we reported that an E. coli strain encoding a Gly-74 to Asp-74 (G74D) thioredoxin mutation could not support wild-type T7 growth and that in vivo, six mutations in T7 gene 5 could individually suppress this G74D thioredoxin defect. In the present study, we report the purification and biochemical characterization of the G74D thioredoxin mutant and two suppressor gene 5 proteins, a Glu-319 to Lys-319 (E319K) mutant of gene 5 protein and an Ala-45 to Thr-45 (A45T) mutant. The suppressor E319K mutation, positioned within the DNA polymerization domain of gene 5 protein, appears to suppress the parental thioredoxin mutation by compensating for the binding defect that was caused by the G74D alteration. We suggest that the Glu 319 residue of T7 gene 5 protein and the Gly-74 residue of E. coli thioredoxin define a contact point or site of interaction between the two proteins. In contrast, the A45T mutation in gene 5 protein, located within the 3' to 5' exonuclease domain, does not suppress the G74D thioredoxin mutation by simple restoration of binding affinity. Based upon our understanding of the mechanisms of suppression, we propose a model for the T7 gene 5 protein-E. coli thioredoxin interaction. PMID- 8702719 TI - Cell-specific expression of the human gonadotropin-releasing hormone gene in transgenic animals. AB - We have previously demonstrated that 1131 base pairs (bp) of the human gonadotropin-releasing hormone (hGnRH) gene promoter can target simian virus 40 T antigen expression to GnRH neurons in transgenic mice. In these animals, GnRH neurons were transformed before they migrated to their final location in the rostral hypothalamus, complicating an analysis of cell-specific expression. To localize regions of the hGnRH promoter that are important for cell-specific expression, we created transgenic mice with various 5'-flanking regions of the hGnRH gene fused to the luciferase reporter gene. When 3828 or 1131 bp of the hGnRH promoter 5'-flanking DNA were used (-3828/+5LUC and -1131/+5LUC, respectively), luciferase expression in adult transgenic mice was observed in the rostral hypothalamus and olfactory tissues, regions which have been shown to be loci of GnRH-expressing neurons. Luciferase expression was not observed in other brain or peripheral tissues. Double-labeled in situ hybridization further demonstrated that luciferase expression was invariably colocalized with GnRH expression. When transgenic animals were created with a construct consisting of 484 bp of the hGnRH 5'-flanking DNA fused to the luciferase gene (-484/+5LUC), luciferase expression was not observed in the hypothalamus or in olfactory tissues. This is the first report localizing DNA sequences responsible for cell specific expression of the GnRH gene in vivo. PMID- 8702720 TI - Cytoplasmic localization of mitogen-activated protein kinase kinase directed by its NH2-terminal, leucine-rich short amino acid sequence, which acts as a nuclear export signal. AB - Mitogen-activated protein kinase (MAPK) is activated in cytoplasm in response to extracellular signals and then is translocated to nucleus. A directed activator for MAPK, MAPK kinase (MAPKK), stays in cytoplasm to transmit the signal from the plasma membrane to MAPK. Here we show that MAPKK contains a short amino acid sequence in the N-terminal region (residues 32-44), which acts as a nuclear export signal (NES) and thus is required for cytoplasmic localization of MAPKK. This NES sequence of MAPKK, like that of protein kinase inhibitor of cAMP dependent protein kinase or Rev, is rich in leucine residues, which are crucial for the NES activity. Furthermore, the NES peptide of protein kinase inhibitor, as well as the NES peptide of MAPKK, inhibited the nuclear export of ovalbumin conjugated to the NES peptide of MAPKK. These results may suggest a common mechanism of nuclear export using a general leucine-rich NES. PMID- 8702718 TI - Identification and characterization of a gene encoding a gut-enriched Kruppel like factor expressed during growth arrest. AB - A cDNA clone, named gut-enriched Kruppel-like factor (GKLF), was isolated from an NIH 3T3 library using a probe encoding the zinc finger region of the immediate early transcription factor zif/268. The deduced GKLF amino acid sequence contains three tandem zinc fingers that are related to members of the Kruppel family of transcription factors. By indirect immunofluorescence, GKLF is localized to the cell nucleus. In cultured fibroblasts, GKLF mRNA is found in high levels in growth-arrested cells and is nearly undetectable in cells that are in the exponential phase of proliferation. The growth-arresting nature of GKLF is demonstrated by an inhibition of DNA synthesis in cells transfected with a GKLF expressing plasmid construct. In the mouse, GKLF mRNA is present in select tissues and is most abundant in the colon, followed by the testis, lung, and small intestine. In situ hybridization experiments indicate that GKLF mRNA is enriched in epithelial cells located in the middle to upper crypt region of the colonic mucosa. Taken together, these results suggest that GKLF is potentially a negative regulator of cell growth in tissues such as the gut mucosa, where cell proliferation is intimately coupled to growth arrest and differentiation. PMID- 8702721 TI - 14-3-3 proteins associate with A20 in an isoform-specific manner and function both as chaperone and adapter molecules. AB - A20, a novel zinc finger protein, is an inhibitor of tumor necrosis factor induced apoptosis. The mechanism by which A20 exerts its protective effect is currently unknown. Several isoforms of the 14-3-3 proteins were found to interact with A20 in a yeast two-hybrid screen. A20 bound several 14-3-3 isoforms in vitro. Moreover, transfected A20 was found to preferentially bind the endogenous eta14-3-3 isoform, whereas the beta/zeta isoforms co-immunoprecipitated much less efficiently, and epsilon14-3-3 had an intermediate affinity. Importantly, c-Raf, a previously described 14-3-3-interacting protein, also preferentially bound the eta isoform. The cellular localization and subcellular fractionation of A20 was dramatically altered by co-transfected 14-3-3, providing the first experimental evidence for the notion that 14-3-3 can function as a chaperone. Furthermore, c Raf and A20 co-immunoprecipitated in a 14-3-3-dependent manner, suggesting that 14-3-3 can function as a bridging or adapter molecule. PMID- 8702722 TI - Interferon-gamma induction of the human leukocyte antigen-E gene is mediated through binding of a complex containing STAT1alpha to a distinct interferon-gamma responsive element. AB - Expression of the human major histocompatibility complex (MHC) class I genes has been shown previously to increase at the transcriptional level following exposure to interferon-gamma (IFN-gamma). In this report we have examined the molecular mechanisms involved in the IFN-gamma-induced transcription of the human MHC class I gene, HLA-E. Functional analysis of CAT reporter gene constructs under the control of the HLA-E promoter transfected into U937 cells revealed the presence of a distinct IFN-gamma-responsive element, termed the interferon response region (IRR), that was necessary and sufficient to mediate the response to IFN-gamma. This cis-acting regulatory sequence contains an imperfect inverted repeat; the 5' half of the IRR resembles the IFN-gamma activation site (GAS), and the 3'-half of the IRR resembles the interferon-stimulated response element (ISRE). Gel mobility shift assays demonstrated that the IRR bound a single, specific, IFN-gamma induced complex (IRR-AC), which was formed rapidly following treatment with IFN gamma and was independent of protein synthesis. Competition experiments with GAS and ISRE sequences from other IFN-inducible genes showed that GAS sequences competed for the IRR-AC, whereas ISRE sequences did not compete. Mutational analysis demonstrated that point mutations in either the 5'-half or 3'-half of the IRR prevented binding of the complex and abrogated or markedly reduced the IFN-gamma responsiveness of reporter gene constructs. Supershift analysis revealed that the IRR-AC contains a factor that was recognized by antibodies specific for the protein STAT1alpha (signal transducer and activator of transcription). Taken together, these findings suggest that the mechanism of IFN gamma-induced transcription of the HLA-E gene is distinct from that of other MHC class I genes. PMID- 8702723 TI - The epidermal growth factor receptor couples transforming growth factor-alpha, heparin-binding epidermal growth factor-like factor, and amphiregulin to Neu, ErbB-3, and ErbB-4. AB - The epidermal growth factor (EGF) family hormones amphiregulin (AR), transforming growth factor-alpha (TGF-alpha), and heparin-binding EGF-like growth factor (HB EGF) are thought to play significant roles in the genesis or progression of a number of human malignancies. However, the ability of these ligands to activate all four erbB family receptors has not been evaluated. Therefore, we have assessed the stimulation of erbB family receptor tyrosine phosphorylation by these hormones in a panel of mouse Ba/F3 cell lines expressing the four erbB family receptors, singly and in pairwise combinations. We also measured the stimulation of interleukin-3-independent survival or proliferation in this panel of Ba/F3 cell lines to compare the patterns of erbB family receptor coupling to physiologic responses induced by these peptides. EGF, TGF-alpha, AR, and HB-EGF all stimulated qualitatively similar patterns of erbB family receptor tyrosine phosphorylation and coupling to physiologic responses. Therefore, EGF, TGF-alpha, AR, and HB-EGF are functionally identical in this model system and behave differently from the EGF family hormones betacellulin and neuregulins. PMID- 8702724 TI - Identification of amino acids in the hormone binding domain of the human estrogen receptor important in estrogen binding. AB - The initial step in the regulation of the transcriptional activity of the estrogen receptor (ER) is the binding of hormone. Previous studies have suggested that the region between amino acids 515 and 535 near the C terminus of the human ER is likely to be important in ligand binding. In order to explicitly define which amino acids in this region are critical for ligand recognition and binding, we have utilized alanine-scanning mutagenesis over the complete 515-535 region. The ability of these 21 mutants to activate transcription in response to the natural estrogen, 17beta-estradiol (E2), was evaluated in cell co-transfection assays with estrogen-responsive reporter genes. In addition, their ability to bind E2 was also tested. Mutations at four sites in the 521-528 region had the greatest effects on E2-induced transcription, with L525A reducing responsiveness 250-fold, G521A and H524A 35-fold, and M528A 11-fold. Mutations at other sites had either no effect or a 4-fold or lesser reduction in sensitivity to E2 (M517A, Y526A, N532A, and P535A). Three of the mutants most affected in their transcriptional response, G521A, H524A, and M528A, showed a coordinate reduction in E2 binding affinity. E2 binding by the most affected mutant, L525A, could not be detected. Thus, the altered transcriptional response of these ER mutants appears to derive solely from an alteration in their affinity for the ligand E2. The four sites most affected by alanine substitution, 521, 524, 525, and 528, follow an alpha-helical periodicity, such that they would be positioned on one face of an alpha-helix. Furthermore, they correspond precisely to residues in an alpha-helix shown to be in contact with ligand in the recently described x-ray crystal structures of two other members of the nuclear hormone receptor superfamily, namely the retinoic acid receptor- and thyroid hormone receptor ligand complexes. Our findings, which broaden observations to the steroid receptor family within the superfamily of nuclear receptors, suggest that this region of the estrogen receptor is in contact with its cognate ligand in a similar fashion. PMID- 8702725 TI - In vitro import of the Rieske iron-sulfur protein by trypanosome mitochondria. AB - Most of the proteins present in the mitochondrion are imported to that location from the cytosol. While this process has been studied extensively in fungal and mammalian systems, little work has been done in other eukaryotic organisms. We are particularly interested in the Trypanosoma brucei system because this organism developmentally regulates mitochondrial function during its life cycle and because one of the imported proteins lacks a conventional targeting sequence. We report here the development of an in vitro import system using crude trypanosome mitochondria and a nuclear encoded, mitochondrial protein. Import of the Rieske iron-sulfur protein subunit of the cytochrome c reductase complex requires a membrane potential, ATP, and a protein component on the mitochondrial surface. The precursor protein is sequentially processed to the mature form in two steps by peptidases that require divalent metal ions for activity. As in other eukaryotic systems, the first processing event occurs inside the inner membrane and is probably catalyzed by a matrix-processing protease. Surprisingly, the second processing activity is located outside the inner membrane. Both processing steps require ATP but are independent of a membrane potential. We suggest that the trypanosome iron-sulfur protein is imported along a "conservative sorting pathway" but that the assembly mechanism of the reductase complex may be unique to trypanosomes. PMID- 8702726 TI - Molecular basis of canine muscle type phosphofructokinase deficiency. AB - Muscle type phosphofructokinase (M-PFK) deficiency is a rare inherited glycogen storage disease in humans that causes exertional myopathy and hemolysis. The molecular basis of canine M-PFK deficiency, the only naturally occurring animal homologue, was investigated. Lack of M-PFK enzyme activity was caused by a nonsense mutation in the penultimate exon of the M-PFK gene, leading to rapid degradation of a truncated (40 amino acids) and therefore unstable M-PFK protein. A polymerase chain reaction-based test was devised to identify M-PFK-deficient and carrier animals. This represents one of only a few inborn errors of metabolism where the molecular defect has been identified in a large animal model which can now be used to develop and assess novel therapeutic strategies. PMID- 8702727 TI - Phosphatidylinositol-3' kinase is not required for mitogenesis or internalization of the Flt3/Flk2 receptor tyrosine kinase. AB - Flt3/Flk2 is a receptor tyrosine kinase that is expressed on early hematopoietic progenitor cells. Flt3/Flk2 belongs to a family of receptors, including Kit and colony-stimulating factor-1R, which support growth and differentiation within the hematopoietic system. The Flt3/Flk2 ligand, in combination with other growth factors, stimulates the proliferation of hematopoietic progenitors of both lymphoid and myeloid lineages in vitro. We report that phosphatidylinositol 3' kinase (PI3K) binds to a unique site in the carboxy tail of murine Flt3/Flk2. In distinction to Kit and colony-stimulating factor-1R, mutant receptors unable to couple to PI3K and expressed in rodent fibroblasts or in the interleukin 3 dependent cell line Ba/F3 provide a mitogenic signal comparable to wild-type receptors. Flt3/Flk2 receptors that do not bind to PI3K also normally down regulate, a function ascribed to PI3K in the context of other receptor systems. These data point to the existence of other unidentified pathways that, alone or in combination with PI3K, transduce these cellular responses following the activation of Flt3/Flk2. PMID- 8702728 TI - Functional importance of amino-terminal domain of Shc for interaction with insulin and epidermal growth factor receptors in phosphorylation-independent manner. AB - Shc has two distinct domains, amino-terminal and SH2 domain, which can interact with activated growth factor receptors. Shc interacts with insulin receptor via Shc-amino-terminal (N) domain, whereas Shc associates with epidermal growth factor (EGF) receptor through both Shc-N and -SH2 domains. In accordance with the different functional roles between insulin and EGF receptors, EGF stimulated tyrosine phosphorylation of Shc faster than insulin. To clarify the functional importance of three distinct Shc domains on insulin and EGF signaling, we microinjected glutathione S-transferase (GST) fusion proteins containing the amino terminus plus collagen homology domain (NCH), collagen homology domain (CH), and Src homology 2 domain (SH2) into Rat1 fibroblasts expressing insulin receptors (HIRc). Bromodeoxyuridine (BrdUrd) incorporation into newly synthesized DNA was subsequently studied to assess the importance of the three distinct domains of Shc. Microinjection of the NCH-GST fusion protein inhibited BrdUrd incorporation induced by both EGF and insulin, whereas microinjection of the SH2 GST fusion protein inhibited EGF, but not insulin stimulation of DNA synthesis. Neither EGF- nor insulin-induced BrdUrd incorporation was inhibited by the CH-GST fusion protein. Following EGF or insulin stimulation, Shc is phosphorylated on single Tyr-317 residue serving as a docking site for Grb2. Microinjection of Shc N+CH GST fusion protein with Tyr-317 --> Phe replacement (Y317F) also inhibited insulin stimulation of DNA synthesis. Next, we stably overexpressed wild-type Shc or Y317F mutant Shc into HIRc cells. Insulin-induced tyrosine phosphorylation of IRS-1 was compared among the transfected cell lines, since IRS-1 and Shc could competitively interact with insulin receptor. Insulin-stimulated tyrosine phosphorylation of IRS-1 was decreased in both WT-Shc and Y317F-Shc cells compared with that in HIRc cells. Furthermore, overexpression of the Shc-SH2 domain or Shc-N+CH domain with Y317F mutation interfered with EGF-stimulated endogenous Shc phosphorylation. These results suggest that the amino terminus domain of Shc is functionally important in insulin- and EGF-induced cell cycle progression and that the phosphorylation of Shc Tyr-317 residue is independent of Shc interaction with these receptors. PMID- 8702729 TI - Replication of O6-methylguanine-containing DNA by repair and replicative DNA polymerases. AB - The biological consequences of O6-methylguanine (m6G) in DNA are well recognized. When template m6G is encountered by DNA polymerases, replication is hindered and trans-lesion replication results in the preferential incorporation of dTMP opposite template m6G. Thus, unrepaired m6G in DNA is both cytotoxic and mutagenic. Yet, cell lines tolerant to m6G in DNA have been isolated, which indicates that some cellular DNA polymerases may replicate m6G-containing DNA with reasonable efficiency. Previous reports suggested that mammalian pol beta could not replicate m6G-containing DNA, but we find that pol beta can catalyze trans-lesion replication; however, the lesion must reside in the optimal context for pol beta activity, single- or short nucleotide gapped substrates. Primed single-stranded DNA templates, with or without template m6G, were poor substrates for pol beta as reported in earlier studies. In contrast, trans-lesion replication by bacteriophage T4 DNA polymerase was observed for primed single stranded DNA templates. Replication of m6G-containing DNA by T4 DNA polymerase required the gp45 accessory protein that clamps the polymerase to the DNA template. The rate-limiting step in replicating m6G-containing DNAs by both DNA polymerases tested was incorporation of dTMP across from the lesion. PMID- 8702730 TI - Purification, cloning, and expression of a novel, endogenous, calcium-sensitive, 28-kDa phosphoprotein. AB - In gastric parietal cells, cholinergically induced increases in intracellular free calcium concentrations have been well characterized, but little is known about the signaling events beyond the initial rise in intracellular calcium. In the present study, we report the isolation of a 28-kDa protein, which is rapidly phosphorylated in intact, enriched parietal cells in response to both the cholinergic agonist, carbachol, and the calcium ionophore, ionomycin. A combination of in situ 32P labeling and one- and two-dimensional gel electrophoresis was used to acquire sufficient quantities of protein to obtain partial amino acid sequence. Cloning of the pp28 cDNA revealed a novel protein which we have named CSPP28 based on its calcium-sensitive phosphorylation. There are three CSPP28 mRNA species (1.7, 2.2, and 3.3 kilobases) that are widely distributed throughout a variety of rabbit tissues. Recombinant CSPP28 was phosphorylated by both crude parietal cell homogenate and purified CaM kinase II in a calcium/calmodulin-dependent manner. We propose that CSPP28 may play an important and ubiquitous role in the calcium signaling pathway. PMID- 8702731 TI - Expression and characterization of a modified flavin-containing monooxygenase 4 from humans. AB - The inability to obtain flavin-containing monooxygenase 4 (FMO4) in heterologous systems has hampered efforts to characterize this isoform of the FMO gene family. Neither the human nor the rabbit ortholog of FMO4, each of which has been cloned and sequenced, has been expressed. Attempts to achieve expression of FMO4 have been made with Escherichia coli, baculovirus, yeast, and COS systems. The cDNAs encoding FMO4 have extended coding regions compared with those encoding other FMO isoforms. The derived amino acid sequences of FMO1, -2, -3, and -5 from all species examined contain about the same number of residues (531-535 residues), whereas the derived sequences of human and rabbit FMO4 contain 558 and 555 residues, respectively. We have investigated whether the elongation of the FMO4 coding region is related to the inability to achieve expression. The cDNA encoding human FMO4 has been modified by a single base change that introduces a stop codon at the consensus position. This modification allows for expression in E. coli. Lack of expression of intact FMO4 is caused by a problem that occurs following transcription, a problem that is overcome completely by relocation of the stop codon 81 bases to 5' of its normal position. Truncated FMO4 is expressed as an active enzyme with characteristics typical of an FMO isoform. Possible functional changes resulting from altering the 3'-end of an FMO were investigated with human FMO3. Elongation of the coding region of the FMO3 cDNA to the next available stop codon (FMO3*) resulted in the expression of an enzyme with properties very similar to those of unmodified FMO3. Elongation of FMO3 lowered the level of expression in E. coli but did not eliminate it. As with FMO4, the difference in expression levels between FMO3 and elongated FMO3 (FMO3*) appears to be related to translation rather than transcription. The functional characteristics of FMO3 and FMO3* are not significantly different. PMID- 8702732 TI - Differential regulation of the stability of cytokine mRNAs in lipopolysaccharide activated blood monocytes in response to interleukin-10. AB - Adenosine-uridine (AU) instability elements, found in the 3'-untranslated regions of numerous mRNAs, target these mRNAs for rapid degradation. In addition, the degradation rate of some mRNAs that contain AU instability elements can change. This modulation of mRNA stability is an important component in the regulation of expression of many of the cytokines that control the production and function of blood cells. However, it has not been clear whether the stabilities of individual cytokine mRNAs that contain AU instability elements are coordinately regulated or whether different mRNAs can be independently regulated. We have investigated the influence of the cytokine synthesis inhibitory factor interleukin (IL)-10 on the turnover of granulocyte-colony stimulating factor (G-CSF), granulocyte macrophage colony stimulating factor (GM-CSF), and IL-10 mRNAs in human blood monocytes stimulated with lipopolysaccharide. We find that all three mRNAs are destabilized in response to IL-10 but at different times. The G-CSF and GM-CSF mRNAs respond similarly, being rapidly destabilized, consistent with a direct influence of IL 10 receptor-mediated signals on the stability of these mRNAs. In contrast the IL 10 mRNA became unstable only after several hours of treatment with IL-10, suggesting that the IL-10 mRNA, although it also contains AU instability elements, is not co-regulated with the G-CSF and GM-CSF mRNAs but is regulated by a secondary factor produced in response to IL-10. PMID- 8702733 TI - Identification of benz(othi)azepine-binding regions within L-type calcium channel alpha1 subunits. AB - To identify the binding domain for diltiazem-like Ca2+ antagonists on L-type Ca2+ channel alpha1 subunits we synthesized the benzazepine [3H]benziazem as a novel photoaffinity probe. [3H]Benziazem reversibly labeled the benzothiazepine (BTZ) binding domain of partially purified skeletal muscle Ca2+ channels with high affinity (Kd = 12 nM) and photoincorporated into its binding domain with high yield (>66%). Antibody mapping of proteolytic labeled fragments revealed specific labeling of regions associated with transmembrane segments S6 in repeats III and IV. More than 50% of the labeling was found in the tryptic fragment alanine 1023 lysine 1077 containing IIIS6 together with extracellular and intracellular amino acid residues. The remaining labeling was identified in a second site comprising segment S6 in repeat IV and adjacent residues. Unlike for dihydropyridines, no labeling was observed in the connecting IIIS5-IIIS6 linker. The [3H]benziazem photolabeled regions must be in close contact to the drug molecule when bound to the channel. We propose that the determinants for high affinity BTZ binding are located within or in close proximity to segments IIIS6 and/or IVS6. Therefore the binding domain for BTZs, like for the other main classes of Ca2+ antagonists, must be located in close proximity to pore-forming regions of the channel. PMID- 8702734 TI - SpFGFR, a new member of the fibroblast growth factor receptor family, is developmentally regulated during early sea urchin development. AB - We describe the cloning of a new fibroblast growth factor receptor, SpFGFR1, that is differentially regulated at the level of transcript abundance during sea urchin embryogenesis. Sequence representing the conserved tyrosine kinase domain was obtained by reverse transcription-polymerase chain reaction using degenerate primers, and the entire open reading frame was obtained by standard cDNA library screening methods. SpFGFR contains a series of domains characteristic of FGFRs: three immunoglobulin-like motifs, an acid box, a transmembrane domain, a relatively long juxtamembrane sequence, a split tyrosine kinase domain, and two conserved intracellular tyrosine residues. Alternative splicing of SpFGFR generates two variants (Ig3L and Ig3S), which differ by insertion in the center of the Ig3 domain of 34 extra amino acids, encoded by an additional exon. Transcripts encoding both variants accumulate when morphogenesis begins with mesenchyme cell ingression and gastrulation. SpFGFR transcripts accumulate in all cell types of the embryo, although in situ hybridization shows that they are somewhat enriched in cells of oral ectoderm and endoderm. Transcripts encoding the Ig3S variant, whose structure resembles more closely that of vertebrate receptors, are enriched in endomesoderm, suggesting that the SpFGFR variants could play distinct roles in the sea urchin embryo. PMID- 8702735 TI - Defining a novel cis-element in the 3'-untranslated region of mammalian ribonucleotide reductase component R2 mRNA. cis-trans-interactions and message stability. AB - Mammalian ribonucleotide reductase is a highly regulated activity essential for DNA synthesis and repair. The 3'-untranslated region (3'-UTR) of mammalian ribonucleotide reductase R2 mRNA has been implicated in the tumor promoter, 12-O tetradecanoylphorbol-13-acetate-mediated stabilization of mouse BALB/c 3T3 R2 message. We investigated the possibility that the 3'-UTR contains regulatory information for R2 mRNA turnover. Using 3'-end-labeled RNA in gel shift and UV cross-linking analyses, we detected in the 3'-UTR a novel 9-nucleotide cis element, 5'-UCGUGUGCU-3', which interacted with a widely distributed cellular cytosolic protease-sensitive factor(s) in a sequence-specific manner to form a 45 kDa R2 binding protein complex. The binding activity was redox-sensitive and down regulated by 12-O-tetradecanoylphorbol-13-acetate and okadaic acid in a dose dependent manner. Insertion of a 154-base pair fragment containing the cis element led to markedly reduced accumulation of chloramphenicol acetyltransferase hybrid mRNA relative to the same insert carrying a series of G --> A mutations within this element that eliminated binding. We suggest that the 9-nucleotide region functions as a destabilizing element. These results provide a model for ribonucleotide reductase gene expression through a novel and specific mRNA cis trans-interaction involving a phosphorylation signal pathway that leads to changes in the stability of R2 message. PMID- 8702736 TI - Soluble adenylyl cyclase from Spodoptera frugiperda (Sf9) cells. Purification and biochemical characterization. AB - An insect ovarian cell, Spodoptera frugiperda (Sf9), has been widely used to express recombinant proteins, including adenylyl cyclase, as a host cell in the baculovirus expression system. We report the presence and characterization of a soluble adenylyl cyclase (sAC) distinct from a membrane-bound form of adenylyl cyclase (mAC) that is also present in Sf9 cells. sAC was purified 3,500-fold to near homogeneity; a single band at 25 kDa on SDS-polyacrylamide gel electrophoresis correlated well with adenylyl cyclase catalytic activity. The purified enzyme had a catalytic activity of 0.1 micromol/min.mg and the Km of 0.55 mM for the substrate ATP. In contrast to mAC, sAC was heat-stable. Enzymatic activity of sAC was not stimulated by forskolin and was inhibited by salts at high concentrations. sAC utilized both manganese- and magnesium-ATP as substrate. Di- or triphosphate-containing nucleotides, such as GTP and GDP, as well as pyrophosphate, noncompetitively inhibited sAC. Our data suggest that the physical and biochemical characteristics of sAC are different from those of mAC in Sf9 cells as well as from those of other known forms of adenylyl cyclase in animal cells; sAC in Sf9 cells may constitute a new member of adenylyl cyclase found in animals. PMID- 8702737 TI - Influence of interleukin-6 (IL-6) dimerization on formation of the high affinity hexameric IL-6.receptor complex. AB - The high affinity interleukin-6 (IL-6) signaling complex consists of IL-6 and two membrane-associated receptor components: a low affinity but specific IL-6 receptor and the affinity converter/signal transducing protein gp130. Monomeric (IL-6M) and dimeric (IL-6D) forms of Escherichia coli-derived human IL-6 and the extracellular ("soluble") portions of the IL-6 receptor (sIL-6R) and gp130 have been purified in order to investigate the effect of IL-6 dimerization on binding to the receptor complex. Although IL-6D has a higher binding affinity for immobilized sIL-6R, as determined by biosensor analysis employing surface plasmon resonance detection, IL-6M is more potent than IL-6D in a STAT3 phosphorylation assay. The difference in potency is significantly less pronounced when measured in the murine 7TD1 hybridoma growth factor assay and the human hepatoma HepG2 bioassay due to time-dependent dissociation at 37 degrees C of IL-6 dimers into active monomers. The increased binding affinity of IL-6D appears to be due to its ability to cross-link two sIL-6R molecules on the biosensor surface. Studies of the IL-6 ternary complex formation demonstrated that the reduced biological potency of IL-6D resulted from a decreased ability of the IL-6D (sIL-6R)2 complex to couple with the soluble portion of gp130. These data imply that IL-6-induced dimerization of sIL-6R is not the driving force in promoting formation of the hexameric (IL-6 IL-6R gp130)2 complex. A model is presented whereby the trimeric complex of IL-6R, gp130, and IL-6M forms before the functional hexamer. Due to its increased affinity for the IL-6R but its decreased ability to couple with gp130, we suggest that a stable IL-6 dimer may be an efficient IL-6 antagonist. PMID- 8702738 TI - Alternative exon splicing controls a translational switch from activator to repressor isoforms of transcription factor CREB during spermatogenesis. AB - The cAMP/protein kinase A signaling pathway activates the cAMP-responsive transcription factor CREB. Here we describe a unique alternative RNA splicing event that occurs during the development of germ cells in the testis, resulting in a translational switch from an mRNA encoding activator CREB to an mRNA encoding novel inhibitor CREB isoforms (I-CREBs). Alternative splicing of an additional exon into the CREB mRNA in mid to late pachytene spermatocytes results in the premature termination of translation and consequent downstream reinitiation of translation producing I-CREBs. The I-CREBs down-regulate cAMP activated gene expression by inhibiting activator CREB from binding to cAMP response elements. Further, the developmental stage-specific expression of I CREBs in germ cells of the seminiferous tubules correlates with the cyclical down regulation of activator CREB, suggesting that I-CREBs repress expression of the cAMP-inducible CREB gene as well as other genes transiently induced by cAMP during the 12-day cycle of spermatogenesis. PMID- 8702739 TI - Expression of recombinant HLA-DR2 molecules. Replacement of the hydrophobic transmembrane region by a leucine zipper dimerization motif allows the assembly and secretion of soluble DR alpha beta heterodimers. AB - Major histocompatibility complex (MHC) class II molecules are membrane-anchored heterodimers that present peptides on the surface of antigen presenting cells to T cells. Soluble HLA-DR2 molecules were expressed for structural and functional characterization of the MHC/peptide/T cell receptor recognition unit. The alpha and beta chains of DR2 (encoded by the DRA, DRB1*1501 genes) did not assemble in mammalian or insect cell lines when the transmembrane regions of one or both chains were truncated. The hydrophobic transmembrane regions of DRalpha and DRbeta facilitate assembly of the heterodimer and were therefore replaced by the leucine zipper dimerization motifs from the transcription factors Fos and Jun, which assemble as a soluble, tightly packed coiled coil structure. The DRalpha Fos and DRbeta-Jun constructs were expressed in a methyltrophic yeast, Pichia pastoris, using the alpha-mating factor secretion signal to direct expression to the secretory pathway. DR alphabeta heterodimers were purified from supernatants using an antibody specific for the DR alphabeta heterodimer. Kinetic and quantitative peptide binding experiments demonstrated that recombinant DR2 molecules were efficiently loaded with an antigenic peptide. Soluble DR2 molecules can be used to define structural aspects of the MHC/peptide/T cell receptor interaction and to study the signals induced by T cell receptor recognition of soluble DR2.peptide complexes. PMID- 8702740 TI - The developmentally regulated avian Ch21 lipocalin is an extracellular fatty acid binding protein. AB - Ch21, a developmentally regulated extracellular protein expressed in chick embryos and in cultured chondrocytes, was expressed in the baculovirus system, and the recombinant protein was purified to homogeneity by gel-filtration chromatography. Separation of two isoforms was achieved on an ion-exchange column. Previous work had shown that Ch21 belongs to the superfamily of lipocalins, which are transport proteins for small hydrophobic molecules. Studies were performed to identify the Ch21 ligand. By analysis of recombinant Ch21 on native polyacrylamide gel electrophoresis and by Lipidex assay, the binding of fatty acid to the protein was shown and a preferential binding of long-chain unsaturated fatty acids was observed. Both isoforms had the same behavior. The binding was saturable. Stoichiometry was about 0.7 mol of ligand/mol of protein. The protein binds the ligand in its monomeric form. Calculated dissociation constants were 2 X 10(-7) M for unsaturated fatty acids and 5 X 10(-7) M for stearic acid. The binding was specific; other hydrophobic molecules, as retinoic acid, progesterone, prostaglandins, and long-chain alcohols and aldehydes did not bind to the protein. Short-chain fatty acids did not bind to the protein. Ch21, also present in chicken serum, represents the first extracellular protein able to selectively bind and transport fatty acid in extracellular fluids and serum. We propose to rename the Ch21 protein as extracellular fatty acid-binding protein (Ex-FABP). PMID- 8702741 TI - A minimal set of RNA polymerase II transcription protein interactions. AB - All pairwise interactions of RNA polymerase II and general transcription factors (TF) IIB, E, F, and H have been quantitated by surface plasmon resonance with the use of a Ni2+ chelate on the sensor surface where necessary to attain higher sensitivity. Only 4 of 10 possible interactions were found above the detection limit: TFIIB, -E, and -F binding to RNA polymerase II and TFIIE binding to TFIIH. These four interactions constitute a minimal set for the formation of a transcription initiation complex and may represent the primary interactions involved in assembly of the complex. Point mutations in TFIIB that alter the location of transcription start sites in vivo markedly diminished the affinity of TFIIB binding to RNA polymerase II. Protein blotting revealed an interaction between the largest subunit of TFIIE and third largest subunit of TFIIH, which may be responsible for TFIIE binding to TFIIH. PMID- 8702742 TI - Differential utilization of Trk autophosphorylation sites. AB - Tyrosine autophosphorylation controls the catalytic and signaling activities of the neurotrophin receptors, the Trks. To analyze the regulation of distinct tyrosine sites, we generated a panel of antibodies that report the phosphorylation state of individual tyrosines within the Trk cytoplasmic domain. Using pheochromocytoma-derived cell lines, we show that individual tyrosines within the nerve growth factor receptor TrkA are phosphorylated in a non coordinate fashion following receptor activation. The non-coordinate response of these tyrosines reflects their separate functions in regulating the catalytic and signaling activities of Trk receptors. The differential utilization of distinct sites on Trk receptor tyrosine kinases suggests that the receptor can specify both the timing and the nature of neurotrophin-stimulated signal transduction pathways. Moreover, we show that these Trk autophosphorylation sites, which have hitherto been mapped and characterized only in non-neuronal cell lines, are activated in normal neurons in response to ligand stimulation. PMID- 8702743 TI - Negative signaling via FcgammaRIIB1 in B cells blocks phospholipase Cgamma2 tyrosine phosphorylation but not Syk or Lyn activation. AB - Crosslinking of the B cell antigen receptor surface immunoglobulin induces tyrosine phosphorylation and activation of the Src family and Syk tyrosine protein kinases, tyrosine phosphorylation of phospholipase Cgamma2 (PLCgamma2) and increases in intracellular second messengers inositol phosphates and Ca2+. These activation events, in conjunction with other pathways, culminate in the induction of B cell proliferation and differentiation. In contrast, co crosslinking surface Ig with the B cell IgG Fc receptor prevents many of these activation events, including B cell proliferation and differentiation. The precise nature of the negative signal(s) derived from Fc receptors that prevent B cell activation is not known. Here, early activation events were examined in B cells stimulated via the antigen receptor alone or under co-crosslinking conditions. The data indicated a selective block in the tyrosine phosphorylation and activation of PLCgamma2 but not in activation of the upstream kinases, Syk and Lyn, under co-crosslinking conditions. We conclude that the negative signal acts directly on PLCgamma2 and is consistent with recent studies describing an activation-induced association of a phosphotyrosine phosphatase with tyrosine phosphorylated B cell Fc receptor. PMID- 8702744 TI - Identification of a Drosophila melanogaster glutamate-gated chloride channel sensitive to the antiparasitic agent avermectin. AB - Glutamate-gated chloride channels, members of the ligand-gated ion channel superfamily, have been shown in nematodes and in insects to be a target of the antiparasitic agent avermectin. Two subunits of the Caenorhabditis elegans glutamate-gated chloride channel have been cloned: GluCl-alpha and GluCl-beta. We report the cloning of a Drosophila melanogaster glutamate-gated chloride channel, DrosGluCl-alpha, which shares 48% amino acid and 60% nucleotide identity with the C. elegans GluCl channels. Expression of DrosGluCl-alpha in Xenopus oocytes produces a homomeric chloride channel that is gated by both glutamate and avermectin. The DrosGluCl-alpha channel has several unique characteristics not observed in C. elegans GluCl: dual gating by avermectin and glutamate, a rapidly desensitizing glutamate response, and a lack of potentiation of the glutamate response by avermectin. The pharmacological data support the hypothesis that the DrosGluCl-alpha channel represents the arthropod H-receptor and an important target for the avermectin class of insecticides. PMID- 8702745 TI - Bcl-2 protects macrophages from nitric oxide-induced apoptosis. AB - Endogenously generated or exogenously supplied nitric oxide (NO)-induced apoptotic cell death in the mouse macrophage cell line RAW 264.7. Apoptotic signaling caused an early accumulation of the tumor suppressor p53 prior to DNA fragmentation. Contrary to the notion of specific activating signals, inhibitory transduction mechanisms largely remain unknown. Therefore, RAW 264.7 macrophages were stably transfected with human Bcl-2, an anti-apoptotic protein. Bcl-2 transfectants showed substantial protection from cell death induced following the exposure to NO donors such as S-nitrosoglutathione (GSNO) and spermine-NO. In contrast, in RAW 264. 7 parent or in neomycin control-transformed cells, these NO donors induced internucleosomal DNA cleavage in a dose-dependent manner. Similarly, expression of the inducible NO synthase in response to lipopolysaccharide and interferon-gamma also caused apoptosis in RAW macrophages and neo controls within 24 h. In contrast, Bcl-2 transfectants appeared highly resistant, although inducible NO synthase levels increased along with concomitant nitrite production similar to control cells. The expression of p53 and Bax was also explored in controls and Bcl-2 transfectants after GSNO addition. GSNO induced p53 expression in Bcl-2 transfectants at levels comparable with nontransfected RAW macrophages. Moreover, GSNO induced increases in the steady state levels of Bax protein in parental and Bcl-2-transfected cells. We conclude therefore, that Bcl-2 acts downstream of p53, presumably nullifying the NO mediated increase in Bax protein in RAW 264.7 cells. PMID- 8702746 TI - The gene 59 protein of bacteriophage T4. Characterization of protein-protein interactions with gene 32 protein, the T4 single-stranded DNA binding protein. AB - The gene 59 protein (gp59) of bacteriophage T4 stimulates the activities of gene 41 protein (gp41), the T4 replicative DNA helicase, by promoting the assembly of gp41 onto single-stranded (ss)-DNA molecules that are covered with cooperatively bound gene 32 protein (gp32). This helicase-ssDNA assembly process, which is important for the reconstitution of the primosome component of the T4 DNA replication fork, appears to require both gp59-gp41 and gp59-gp32 protein-protein interactions. In this study we characterize the physical and functional interactions of gp59 with gp32, the T4 ssDNA-binding protein. Experimental results presented herein indicate: 1) that gp59 binds specifically to both free and ssDNA-bound gp32 molecules; and 2) that in both cases binding involves contacts between gp59 and the acidic C-terminal domain of gp32 (the so-called "A domain"). We further show that single-stranded DNA molecules coated with (gp32 A), a truncated form of gp32 lacking the A-domain, are refractory to gp59 dependent helicase assembly. The data indicate that specific contacts between gp59 molecules and the A-domains of gp32 molecules are essential for gp59 dependent assembly of gp41 onto gp32-ssDNA complexes. Our results are consistent with a model in which gp59 binds to gp32 molecules within the gp32-ssDNA complex and therein forms a target site for helicase-ssDNA assembly. PMID- 8702747 TI - A C-terminal mutant of the G protein beta subunit deficient in the activation of phospholipase C-beta. AB - The molecular mechanism by which the G protein betagamma complex modulates multiple mammalian effector pathways is unknown. Homolog-scanning mutagenesis of the G protein beta subunit was employed to identify residues critical for the activation of phospholipase C-beta2 (PLC-beta2). A series of chimeras was made by introducing small segments of the Dictyostelium beta subunit into a background of mammalian beta1 and tested in COS cell cotransfection assays for their ability to activate PLC-beta2 and assemble with mammalian gamma2. A chimera that contained four Dictyostelium beta substitutions within the C-terminal 14 residues was unable to activate PLC-beta2 when cotransfected with gamma, despite its demonstrable expression in a gamma-dependent manner. Cotransfection of the mutant blocked m2 muscarinic receptor activation of PLC by a pertussis toxin-sensitive pathway. This C-terminal mutant retained the ability, however, to stimulate the mitogen-activated protein kinase pathway. These results imply that activation of different betagamma-responsive effectors is mediated by distinct domains. PMID- 8702748 TI - Purification, characterization, and cDNA cloning of a galactose-specific C-type lectin from Drosophila melanogaster. AB - We purified a lectin from a pupal extract of Drosophila melanogaster. This lectin agglutinated trypsinized and glutaraldehyde-fixed bovine red blood cells in the presence of calcium or magnesium. The hapten sugar of this lectin was galactose. The molecular mass of the intact lectin was determined to be 41 kDa, and it comprised 14- and 17-kDa subunits. The 17-kDa subunit was shown to be a glycosylated form of the 14-kDa subunit. Analysis of the cDNA for this lectin revealed that the 14-kDa subunit consists of 163 amino acid residues and contains all residues conserved in various C-type lectins. It was suggested that the Drosophila lectin and Sarcophaga lectin share some properties and function similarly in defense and development, but probably they are not structural homologues. PMID- 8702749 TI - Cell cycle regulation of membrane phospholipid metabolism. PMID- 8702750 TI - N-Ethylmaleimide-sensitive factor acts at a prefusion ATP-dependent step in Ca2+ activated exocytosis. AB - An ATP-dependent activity of NSF (N-ethylmaleimide-sensitive factor) that rearranges soluble NSF attachment protein (SNAP) receptor (SNARE) protein complexes was proposed to be the driving force for membrane fusion. The Ca2+ activated fusion of secretory vesicles with the plasma membrane in permeable PC12 cells requires ATP; however, the ATP requirement is for a priming step that precedes the Ca2+-triggered fusion reaction. While phosphoinositide phosphorylation is a key reaction required for priming, additional ATP-dependent reactions are also necessary. Here we report that the NSF-catalyzed rearrangement of SNARE protein complexes occurs during ATP-dependent priming. NSF with alpha SNAP (soluble NSF attachment protein) were required for ATP-dependent priming but not Ca2+-triggered fusion, indicating that NSF acts at an ATP-dependent prefusion step rather than at fusion itself. NSF-catalyzed activation of SNARE proteins may reorganize membranes to generate a vesicle-plasma membrane prefusion intermediate that is poised for conversion to full fusion by Ca2+-dependent mechanisms. PMID- 8702751 TI - SNAP-25 is required for a late postdocking step in Ca2+-dependent exocytosis. AB - The Ca2+-activated fusion of large dense core vesicles (LDCVs) with the plasma membrane is reconstituted in mechanically permeabilized PC12 cells by provision of millimolar MgATP and cytosolic proteins. Ca2+-activated LDCV exocytosis was inhibited completely by the type E but not the type A botulinum neurotoxin (BoNT) even though both BoNTs were equally effective in proteolytically cleaving the synaptosome-associated protein of 25 kDa (SNAP-25). The greater inhibition of exocytosis by BoNT E correlated with a greater destabilization of detergent extracted complexes consisting of SNAP-25, synaptobrevin, and syntaxin. LDCVs in permeable PC12 cells can be poised at a late postdocking, prefusion state by MgATP-dependent priming processes catalyzed by N-ethylmaleimide sensitive factor and priming in exocytosis proteins. BoNT E completely blocked Ca2+-activated LDCV exocytosis in ATP-primed cells, whereas BoNT A was only slightly inhibitory, implying that the C-terminal region of SNAP-25 (Ile181-Gln197) between the cleavage sites for BoNT E and BoNT A is essential for late postdocking steps. A required role for SNAP-25 at this stage was also indicated by inhibition of Ca2+ activated LDCV fusion in ATP-primed cells by a C-terminal peptide antibody. We conclude that plasma membrane SNAP-25, particularly residues 181-197, is required for Ca2+-regulated membrane fusion at a step beyond LDCV docking and ATP utilization. PMID- 8702752 TI - Molecular cloning and characterization of the human anaphylatoxin C3a receptor. AB - In a human neutrophil cDNA library, an orphan G-protein-coupled receptor, HNFAG09, with 37% nucleotide identity to the C5a receptor (C5a-R, CD88) was identified. A novel feature of this gene, unlike C5a-R and other G-protein coupled receptors, is the presence of an extraordinarily large predicted extracellular loop comprised of in excess of 160 amino acid residues between transmembrane domains 4 and 5. Northern blot analysis revealed that expression of mRNA for this receptor in human tissues, while similar, was distinct from C5a-R expression. Although there were differences in expression, transcripts for both receptors were detected in tissues throughout the body and the central nervous system. Mammalian cells stably expressing HNFAG09 specifically bound 125I-C3a and responded to a C3a carboxyl-terminal analogue synthetic peptide and to human C3a but not to rC5a with a robust calcium mobilization response. HNFAG09 encodes the human anaphylatoxin C3a receptor. PMID- 8702753 TI - p62, a phosphotyrosine-independent ligand of the SH2 domain of p56lck, belongs to a new class of ubiquitin-binding proteins. AB - p62 is a novel cellular protein which was initially identified as a phosphotyrosine-independent ligand of the SH2 domain of p56(lck). In the yeast two-hybrid system, p62 specifically interacted with ubiquitin in vivo. Furthermore, p62 bound to ubiquitin-conjugated Sepharose beads in vitro and was efficiently competed by soluble ubiquitin. The interaction was independent of ATP hydrolysis, and its dissociation did not require a reducing agent. Thus, p62 binds to ubiquitin noncovalently. Further analysis showed that the C-terminal 80 amino acids of p62 were indispensable for its interaction with ubiquitin. However, p62 has homology neither with ubiquitin C-terminal hydrolases nor with the S5a subunit of the 26 S proteasome complex, the only proteins known to bind to ubiquitin noncovalently. These results suggest that p62 belongs to a new class of ubiquitin-binding proteins and that p62 affects signal transduction at least partly through ubiquitination-mediated protein degradation. PMID- 8702754 TI - Molecular characterization and in vitro biological activity of placentin, a new member of the insulin gene family. AB - Insulin and insulin-like growth factors belong to a family of polypeptides involved in essential physiological processes. Placentin, a new member of the insulin family, was recently identified as a 139-amino acid open reading frame from a cDNA clone isolated from a subtracted library of first trimester human placenta. Tris/Tricine/SDS-polyacrylamide gel electrophoresis and immunoblot analyses of histidine-tagged recombinant placentin indicate that it is composed of two peptide chains of apparent molecular masses of 4 and 13 kDa. Conditioned media produced by recombinant expression of placentin cDNA in the placental 3AsubE cell line were assayed for biological activity and found to stimulate tyrosine phosphorylation and DNA synthesis. While these effects closely mimicked those of insulin, they were not mediated by the insulin receptor as shown by the lack of tyrosine phosphorylation of this receptor upon placentin treatment. Moreover, in cytotrophoblast primary culture, production of chorionic gonadotropin, a marker of trophoblast differentiation, was increased upon treatment with placentin-conditioned media, while unaffected by insulin. These results suggest that placentin might participate in the cellular proliferation and/or differentiation processes during placental development. PMID- 8702755 TI - Two yeast homologs of ECA39, a target for c-Myc regulation, code for cytosolic and mitochondrial branched-chain amino acid aminotransferases. AB - ECA39 was isolated as a target gene for c-Myc regulation in mice. We identified two homologs for the murine ECA39 in the yeast Saccharomyces cerevisiae, ECA39 and ECA40, as well as two human homologs. These genes show a significant homology to prokaryotic branched-chain amino acid aminotransferase (BCAT) (EC). To understand the function of eukaryotic ECA39 and ECA40, we deleted either gene from the yeast genome. Activity of branched-chain amino acid aminotransferase was measured in the wild-type and mutants with either leucine, isoleucine, or valine as substrates. The results demonstrate that in S. cerevisiae ECA39 and ECA40 code for mitochondrial and cytosolic branched-chain amino acid aminotransferases, respectively. ECA39 is highly expressed during log phase and is down-regulated during the stationary phase of growth, while ECA40 shows an inverse pattern of gene expression. In agreement with these results, while we previously showed that deletion of ECA39 affected the cell cycle in proliferating cells, we do not observe a growth phenotype in eca40Delta cells. We suggest that BCAT is a target for c-Myc activity and discuss the evolutionary conservation of prokaryotic and eukaryotic BCATs and their possible involvement in regulation of cell proliferation. PMID- 8702756 TI - Phosphorylation and activation of myosin by Rho-associated kinase (Rho-kinase). AB - The small GTPase Rho is implicated in physiological functions associated with actin-myosin filaments such as cytokinesis, cell motility, and smooth muscle contraction. We have recently identified and molecularly cloned Rho-associated serine/threonine kinase (Rho-kinase), which is activated by GTP Rho (Matsui, T., Amano, M., Yamamoto, T., Chihara, K., Nakafuku, M., Ito, M., Nakano, T., Okawa, K., Iwamatsu, A., and Kaibuchi, K. (1996) EMBO J. 15, 2208-2216). Here we found that Rho-kinase stoichiometrically phosphorylated myosin light chain (MLC). Peptide mapping and phosphoamino acid analyses revealed that the primary phosphorylation site of MLC by Rho-kinase was Ser-19, which is the site phosphorylated by MLC kinase. Rho-kinase phosphorylated recombinant MLC, whereas it failed to phosphorylate recombinant MLC, which contained Ala substituted for both Thr-18 and Ser-19. We also found that the phosphorylation of MLC by Rho kinase resulted in the facilitation of the actin activation of myosin ATPase. Thus, it is likely that once Rho is activated, then it can interact with Rho kinase and activate it. The activated Rho-kinase subsequently phosphorylates MLC. This may partly account for the mechanism by which Rho regulates cytokinesis, cell motility, or smooth muscle contraction. PMID- 8702757 TI - The unpredicted high affinities of a large number of naturally occurring tachykinins for chimeric NK1/NK3 receptors suggest a role for an inhibitory domain in determining receptor specificity. AB - Three chimeric receptors were constructed by exchanging exon sequences between human NK1 and NK3 receptor genes. The resulting chimeric receptors not only retained high affinities for their natural ligands substance P and neurokinin B but also exhibited surprisingly high affinities for other naturally occurring tachykinins including neurokinin A, neuropeptide K, neuropeptide gamma, eledoisin, kassinin, physalaemin, and phyllomedusin. In contrast, these chimeric receptors displayed a wide range of variability in their affinities for non naturally occurring ligands including selective agonists and antagonists of NK1, NK2, and NK3 receptors. Since the only common feature among these naturally occurring neurokinin peptides is the conserved C-terminal sequences, our data suggest that these conserved sequences must play the major role in conferring high affinity binding to the chimeric receptors. To explain the apparently "improved" affinities of these naturally occurring ligands for the chimeric receptors as compared with their affinities for the parent NK1 and NK3 receptors, we are proposing that certain inhibitory domains that are present in the NK1 and/or NK3 receptors are compromised in these chimeric receptors. Upon disruption of these inhibitory domains during the formation of chimeras, the naturally occurring ligands can interact more favorably with chimeric receptors through their conserved C-terminal sequences. Based on this hypothesis, the binding affinities of natural tachykinin ligands may be largely determined by their conserved C-terminal sequences, whereas receptor selectivities of these ligands are influenced more by the presence or absence of inhibitory domains rather than specific binding domains on their target receptors. PMID- 8702758 TI - Effects of integration host factor and DNA supercoiling on transcription from the ilvPG promoter of Escherichia coli. AB - Integration host factor (IHF) activates transcription from the ilvPG promoter by severely distorting the DNA helix in an upstream region of a supercoiled DNA template in a way that alters the structure of the DNA in the downstream promoter region and facilitates open complex formation. In this report, the in vivo and in vitro influence of DNA supercoiling on transcription from this promoter is examined. In the absence of IHF, promoter activity increases with increased DNA supercoiling. In the presence of IHF, the same increases in superhelical DNA densities result in larger increases in promoter activity until a maximal activation of 5-fold is obtained. However, the relative transcriptional activities of the promoter in the presence and absence of IHF at any given DNA superhelical density remains the same. Thus, IHF and increased DNA supercoiling activate transcription by different mechanisms. Also, IHF binds with equal affinities to its target site on linear and supercoiled DNA templates. Therefore, IHF binding does not activate transcription simply by increasing the local negative supercoiling of the DNA helix in the downstream promoter region or by differential binding to relaxed and supercoiled DNA templates. PMID- 8702759 TI - Characterization of cis-acting elements of the gene for macrophage-stimulating protein from the human. The involvement of positive and negative regulatory elements. AB - To analyze the promotor region of the human macrophage-stimulating protein (MSP) gene, the 5'-flanking region of this gene was cloned. The major initiation site was determined at T located 49 base pairs upstream of the translation initiation site by primer extention with mRNA from HepG2 and Hep3B cells. There was no TATA sequence in this region. Transient transfection assay with 5'-deletion constructs showed that the transcription of this gene was regulated by positive and negative regulatory elements (PRE and NRE). The PRE (-34 to +2) was essential for the maximal transcription of this gene, and the NRE (-141 to -34) appeared to be responsible for the tissue-specific expression of the gene. The PRE contained the CCAAT sequence and a mutation from CCAAT to CTGAT resulted in a significant loss of the transcriptional activity. Electrophoretic mobility shift assay suggested that two different proteins bound to the PRE (MSP-PRE-binding protein-1 (MSP PREB1) and 2). MSP-PREB1 and 2 were detected in various cell types, and the CCAAT sequence was involved in these bindings. These findings indicate that MSP-PREB1 and 2 are positive regulators. Further characterization also revealed that MSP PREB2 was identical to CCAAT binding transcription factor, also known as NF-Y. PMID- 8702760 TI - Regulation of membrane and subunit interactions by N-myristoylation of a G protein alpha subunit in yeast. AB - Initiation of the mating process in yeast Saccharomyces cerevisiae requires the action of secreted pheromones and G protein-coupled receptors. As in other eukaryotes, the yeast G protein alpha subunit undergoes N-myristoylation (GPA1 gene product, Gpa1p). This modification appears to be essential for function, since a myristoylation site mutation exhibits the null phenotype in vivo (gpa1(G2A)). Here we examine how myristoylation affects Gpa1p activity in vitro. We show that the G2A mutant of Gpa1p, when fused with glutathione S-transferase, can still form a complex with the G protein betagamma subunits. The complex is stabilized by GDP and is dissociated upon treatment with guanosine 5'-O (thiotriphosphate). In addition, there is no apparent difference in the relative binding affinity of Gbetagamma for mutant and wild-type Gpa1p. Using sucrose density gradient fractionation of cell membranes, Gpa1p associates normally with the plasma membrane whereas Gpa1pG2A is mislocalized to a microsomal membrane fraction. A portion of Gbetagamma is also mislocalized in these cells, as it is in a gpa1Delta strain. In contrast, wild-type Gpa1p reaches the plasma membrane in cells that do not express Gbetagamma or cell surface receptors. These findings indicate that mislocalization of Gpa1pG2A is not caused by a redistribution of Gbetagamma, nor is it the result of any difference in Gbetagamma binding affinity. These data suggest that myristoylation is required for specific targeting of Gpa1p to the plasma membrane, where it is needed to interact with the receptor and to regulate the release of Gbetagamma. PMID- 8702761 TI - ATP synthase of yeast mitochondria. Isolation of the subunit h and disruption of the ATP14 gene. AB - A new subunit of the yeast ATP synthase (termed subunit h) has been isolated. Amino acid composition and N-terminal sequencing were determined by chemical methods. These data were in agreement with the sequence of the hypothetical protein L8003.20 whose primary structure was deduced from DNA sequencing of the yeast chromosome XII. The amino acid sequence encoded by ATP14 gene is 32 amino acids longer than the mature protein, which contains 92 amino acids corresponding to a calculated mass of 10,408 Da. The protein is hydrophilic and acidic with a calculated pHi of 4.08. It is not apparently related to any subunit described in other ATP synthases. A null mutant was constructed. The mutation was recessive and the mutant strain was unable to grow on glycerol medium. A high percentage of rho- cells arose spontaneously. The mutant mitochondria had no detectable oligomycin-sensitive ATPase activity, but still contained ATPase activity with a catalytic sector dissociated from the membranous components. The mutant mitochondria did not contain subunit h, and the mitochondrially encoded hydrophobic subunit 6 was not present. PMID- 8702762 TI - The relationship of intracellular iron chelation to the inhibition and regeneration of human ribonucleotide reductase. AB - The depletion of cellular iron can lead to the inhibition of ribonucleotide reductase, preventing new DNA synthesis and hence inhibiting cell proliferation. Electron paramagnetic resonance (EPR) spectroscopy has been used to examine simultaneously for the first time the relationship between chelation of intracellular iron and the rate of removal and regeneration of the tyrosyl radical of ribonucleotide reductase within intact human leukemia K562 cells. The different physiochemical characteristics of relatively hydrophobic low molecular weight bidentate hydroxypyridinone chelators and the higher molecular weight hexadentate ferrioxamine have been exploited to elucidate these interactions further. The base-line concentration of EPR-detectable mononuclear nonheme iron complexes was 3.15 =/- 1.05 microM, rising on incubation with chelators more rapidly with hydroxypyridinones than with desferrioxamine. Hydroxypyridinones also removed the tyrosyl radical more rapidly, apparently as a consequence of depletion of the intracellular iron pools necessary to regenerate the active enzyme and compatible with their reportedly greater cell toxicity. The radical decay rate is consistent with previous models, suggesting that iron is spontaneously removed from mammalian ribonucleotide reductase. Upon removal of extracellular chelator the regeneration of the tyrosyl radical was significantly faster for hydroxypyridinones than for desferrioxamine, consistent with their differential effects on cell cycle synchronization. PMID- 8702763 TI - The Yarrowia lipolytica gene PAY5 encodes a peroxisomal integral membrane protein homologous to the mammalian peroxisome assembly factor PAF-1. AB - Pay mutants of the yeast Yarrowia lipolytica fail to assemble functional peroxisomes. One mutant strain, pay5-1, lacks normal peroxisomes and instead contains irregular vesicular structures surrounded by multiple unit membranes. The pay5-1 mutant is not totally deficient in peroxisomal matrix protein targeting, as a subset of matrix proteins continues to localize to a subcellular fraction enriched for peroxisomes. The functionally complementing gene PAY5 encodes a protein, Pay5p, of 380 amino acids (41,720 Da). Pay5p is a peroxisomal integral membrane protein homologous to mammalian PAF-1 proteins, which are essential for peroxisome assembly and whose mutation in humans results in Zellweger syndrome. Pay5p is targeted to mammalian peroxisomes, demonstrating the evolutionary conservation of the targeting mechanism for peroxisomal membrane proteins. Our results suggest that in pay5 mutants, normal peroxisome assembly is blocked, which leads to the accumulation of the membranous vesicular structures observed. PMID- 8702764 TI - Mutations in the PAY5 gene of the yeast Yarrowia lipolytica cause the accumulation of multiple subpopulations of peroxisomes. AB - We previously reported the cloning of the PAY5 gene of the yeast Yarrowia lipolytica by complementation of the peroxisome assembly mutant pay5-1 (Eitzen, G. A., Titorenko, V. I., Smith, J. J., Veenhuis, M., Szilard, R. K., and Rachubinski, R. A. (1996) J. Biol. Chem. 271, 20300-20306). The peroxisomal integral membrane protein Pay5p is a homologue of mammalian PAF-1 proteins, which are essential for peroxisome assembly and whose mutation in humans results in peroxisome biogenesis disorders. Mutations in the PAY5 gene result in the accumulation of three distinct peroxisomal subpopulations. These subpopulations are characterized by differences in 1) buoyant density, 2) the relative distribution of peroxisomal matrix and membrane proteins, 3) the efficiency of import of several peroxisomal matrix proteins, and 4) the phospholipid levels of peroxisomal membranes. These data, together with the analysis of temporal changes in the relative abundance of individual peroxisomal subpopulations in pay5 mutants, suggest that these subpopulations represent intermediates in a multistep peroxisome assembly pathway normally operating in yeast cells. PMID- 8702765 TI - A molecular basis for affinity modulation of Fab ligand binding to integrin alphaIIb beta3. AB - The Arg-Gly-Asp (RGD) sequence within the third complementarity-determining region (CDR3) of the heavy chain (H3) is responsible for the binding of the recombinant murine Fab molecules, AP7 and PAC1.1, to the platelet integrin alphaIIbbeta3. AP7 binding is minimally influenced by the conformational state of this receptor, whereas PAC1.1 binds preferentially to the activated state of the receptor induced by platelet agonists. To study the molecular basis for this functional difference, we replaced the AP7 H3 loop (HPFYRGDGGN) with all or segments of the analogous sequence from PAC1.1 (RSPSYYRGDGAGP). AP7 Fd (VH domain + Cgamma1 domain) segments containing these H3 loop sequences were expressed as active Fab molecules by coinfection of Spodoptera frugiperda cell lines with recombinant baculoviruses containing Fd and AP7 kappa chain cDNA. Replacement of the entire AP7 H3 loop with that from PAC1.1 generated the mutant AP7.3 Fab molecule, which bound selectively to either activated, gel-filtered platelets or to purified alphaIIbbeta3 in a manner identical to that of PAC1.1. Identical results were obtained when solely the sequences flanking the amino side of RGD within the respective H3 loops were exchanged. AP7.3 and PAC1.1 exhibited saturable but submaximal binding to activated gel-filtered platelets. Relative to AP7, the number of AP7.3 or PAC1. 1 Fab molecules bound per platelet was 17% in the presence of 1 m Ca2+ + 1 mM Mg2+ or 40% in the presence of 10 microM Mn2+. The ratio of Fab molecules bound after versus before activation (mean =/- S.D.; n = 3) was: for AP7.3, 9.8 =/- 0.6; for PAC1.1, 8.8 +/- 0.3; and for AP7, 1.4 =/- 0.2. In addition, AP7 bound to the stably expressed integrin mutant alphaIIbbeta3(S123A), whereas AP7.3 and PAC1 did not. Because AP7.3 behaves in every respect like PAC1.1, we conclude that the ability of RGD-based ligands to distinguish activated from resting conformations of the integrin alphaIIbbeta3 can be regulated by limited amino acid sequences immediately adjacent to the RGD tripeptide. Furthermore, those Fab molecules that exhibit increased selectivity for the activated conformation of alphaIIbbeta3 bind to a subpopulation of this integrin on platelets that is modulated by divalent cations. PMID- 8702766 TI - Crystal structure of L-2-haloacid dehalogenase from Pseudomonas sp. YL. An alpha/beta hydrolase structure that is different from the alpha/beta hydrolase fold. AB - L-2-Haloacid dehalogenase catalyzes the hydrolytic dehalogenation of L-2 haloalkanoic acids to yield the corresponding D-2-hydroxyalkanoic acids. The crystal structure of the homodimeric enzyme from Pseudomonas sp. YL has been determined by a multiple isomorphous replacement method and refined at 2.5 A resolution to a crystallographic R-factor of 19.5%. The subunit consists of two structurally distinct domains: the core domain and the subdomain. The core domain has an alpha/beta structure formed by a six-stranded parallel beta-sheet flanked by five alpha-helices. The subdomain inserted into the core domain has a four helix bundle structure providing the greater part of the interface for dimer formation. There is an active site cavity between the domains. An experimentally identified nucleophilic residue, Asp-10, is located on a loop following the amino terminal beta-strand in the core domain, and other functional residues, Thr-14, Arg-41, Ser-118, Lys-151, Tyr-157, Ser-175, Asn-177, and Asp-180, detected by a site-directed mutagenesis experiment, are arranged around the nucleophile in the active site. Although the enzyme is an alpha/beta-type hydrolase, it does not belong to the alpha/beta hydrolase fold family, from the viewpoint of the topological feature and the position of the nucleophile. PMID- 8702767 TI - Identification of ligand binding determinants in the somatostatin receptor subtypes 1 and 2. AB - The somatostatin (SRIF) receptors (SSTRs) 1 and 2 bind SRIF and SRIF 28 with high affinity, although a number of synthetic hexapeptide and octapeptide analogs of SRIF bind selectively to SSTR2. Extracellular loop three and its adjoining trans membrane-spanning regions contain elements essential for the binding of such analogs to murine SSTR2. In particular, a stretch of amino acids from residues 294-297 (FDFV) in murine SSTR2 in trans-membrane domain seven can determine affinity for the SSTR2-selective analogs. Within this region, Phe294 has previously been predicted to be essential for the binding of octapeptides (Kaupmann, K., Bruns, C., Raulf, F., Weber, H., Mattes, H., and Lubbert, H. (1995) EMBO J. 14, 727-735) based on the observation that SSTR1 can bind the octapeptide SMS-201-995 with reasonable affinity after a Ser-to-Phe conversion in the analogous region of this receptor (SSTR1S305F). We find that SSTR1S305F has low affinity for a number of SSTR2-selective hexapeptides, suggesting that these analogs have different binding requirements than SMS-201-995. A correlation is seen between the ability of SSTR1S305F to bind hexapeptide analogs and the presence of a phenylalanine, but not tyrosine, at position two in these small cyclic molecules. Thus, a single hydroxyl group in hexapeptides can play a critical role in determining receptor binding to these receptor mutants. We also find that the second extracellular loop of SSTR1 is important for the selectivity of certain SRIF agonists for binding to SSTR1. Taken together, our data indicate that there are multiple elements in the somatostatin receptors that can determine the binding affinity and selectivity of peptide analogs. PMID- 8702768 TI - Membrane topography and near-neighbor relationships of the mitochondrial ATP synthase subunits e, f, and g. AB - The well characterized subunits of the bovine ATP synthase complex are the alpha, beta, gamma, delta, and epsilon subunits of the catalytic sector, F1; the ATPase inhibitor protein; and subunits a, b, c, and d, OSCP (oligomycin sensitivity conferring protein), F6, and A6L, which are present in the membrane sector, F0, and the 45-A-long stalk that connects F1 to F0. It has been shown recently that bovine ATP synthase preparations also contain three small polypeptides, designated e, f, and g, with respective molecular masses of 8.2, 10. 2, and 11.3 kDa. To ascertain their involvement as bona fide subunits of the ATP synthase and to investigate their membrane topography and proximity to the above ATP synthase subunits, polyclonal antipeptide antibodies were raised in the rabbit to the COOH terminal amino acid residues 57-70 of e, 75-86 of f, and 91-102 of g. It was shown that (i) e, f, and g could be immunoprecipitated with anti-OSCP IgG from a fraction of bovine submitochondrial particles enriched in oligomycin-sensitive ATPase; (ii) the NH2 termini of f and g are exposed on the matrix side of the mitochondrial inner membrane and can be curtailed by proteolysis; (iii) the COOH termini of all three polypeptides are exposed on the cytosolic side of the inner membrane; and (iv) f cross-links to A6L and to g, and e cross-links to g and appears to form an e-e dimer. Thus, the bovine ATP synthase complex appears to have 16 unlike subunits, twice as many as its counterpart in Escherichia coli. PMID- 8702769 TI - The AF-2 region of the retinoic acid receptor alpha mediates retinoic acid inhibition of estrogen receptor function in breast cancer cells. AB - The growth of estrogen receptor (ER)-positive breast cancer cells is inhibited by all-trans-retinoic acid (RA). In the present study, estrogen (E2) induction of pS2 mRNA levels was significantly reduced within 6 h following cotreatment with RA. In transient transfection experiments, RA repressed transactivation from a vitellogenin E2-responsive element by approximately 50% and wild-type RA receptor alpha (RARalpha) or RARbeta enhanced this inhibition. Transfection of truncated RARalpha mutants terminating before or at amino acid 412 markedly decreased RA inhibition of E2-induced reporter gene activity. Expression of RARs with deletions of amino acids 413 and 414 in the transactivation-2 (AF-2) domain also reduced RA inhibition, while deletions and point mutations beyond amino acid 414 behaved like the wild-type RARalpha. RA-treated MCF-7 cells transfected with an RARalpha AF-2 region mutant were twice as sensitive to growth inhibition as untransfected and vector-transfected control cells. Thus, the AF-2 domain in the C terminus of the RARalpha mediates RA inhibition of ER-induced transcription in breast cancer cells. In addition, transcriptional interference between RARs and ERs may contribute to RA inhibition of ER-positive breast cancer cell growth. PMID- 8702770 TI - Structural determinants of the specificity for synaptic vesicle-associated membrane protein/synaptobrevin of tetanus and botulinum type B and G neurotoxins. AB - Tetanus and botulinum neurotoxins type B and G are zinc-endopeptidases of remarkable specificity. They recognize and cleave a synaptic vesicle-associated membrane protein (VAMP)/synaptobrevin, an essential protein component of the vesicle docking and fusion apparatus. VAMP contains two copies of a nine-residue motif, also present in SNAP-25 (synaptosomal-associated protein of 25 kDa) and syntaxin, the two other substrates of clostridial neurotoxins. This motif was suggested to be a determinant of the target specificity of neurotoxins. Antibodies raised against this motif cross-react among VAMP, SNAP-25, and syntaxin and inhibit the proteolytic activity of the neurotoxins. Moreover, the various neurotoxins cross-inhibit each other's proteolytic action. The role of the three negatively charged residues of the motif in neurotoxin recognition was probed by site-directed mutagenesis. Substitution of acidic residues in both copies of the VAMP motif indicate that the first one is involved in tetanus neurotoxin recognition, whereas the second one is implicated in binding botulinum B and G neurotoxins. These results suggest that the two copies of the motif have a tandem association in the VAMP molecule. PMID- 8702771 TI - The cell-binding domain of intimin from enteropathogenic Escherichia coli binds to beta1 integrins. AB - Bacteria interact with mammalian cells surface molecules, such as integrins, to colonize tissues and evade immunological detection. Herein, the ability of intimin, an outer membrane protein from enteropathogenic Escherichia coli, to bind beta1 integrins was investigated. Solid-phase binding assays revealed binding of the carboxyl-terminal 280 amino acids of intimin (Int280) to alpha4beta1 and alpha5beta1 integrins. The binding required divalent ions (in particular, it was enhanced by Mn2+) and was inhibited by an RGD-containing peptide. Nonderivatized Int280, but not Int280CS (like Int280 but with Cys-937 replaced by Ser) blocked the binding of biotinylated Int280 to integrins. Int280 did not efficiently inhibit beta1 integrin binding of invasin from Yersinia pseudotuberculosis. Both intimin and invasin, immobilized on plastic surfaces, mediated adherence of resting or phorbol 12-myristate 13-acetate-activated human CD4(+) T cells, whereas fibronectin mediated the adherence of only activated T cells. T cell binding to intimin and invasin was integrin mediated because it was specifically blocked by an RGD-containing peptide and by antibodies directed against the integrin subunits beta1, alpha4, and alpha5. These results demonstrate a specific integrin binding activity for intimin that is related to, but distinct from, that of invasin. PMID- 8702772 TI - The inhibitory anti-beta1 integrin monoclonal antibody 13 recognizes an epitope that is attenuated by ligand occupancy. Evidence for allosteric inhibition of integrin function. AB - Integrin-ligand binding causes conformational changes in the integrin, as evidenced by the increased expression of epitopes known as ligand-induced binding sites. Some monoclonal antibodies (mAbs) that recognize ligand-induced binding sites stimulate ligand binding, possibly by stabilizing the ligand-occupied conformation of the integrin. Here we have investigated the effect of ligand recognition by alpha5beta1 on the binding of a mAb that inhibits beta1 integrin function (mAb 13). Ligand (fibronectin fragment or GRGDS peptide) decreased the binding of mAb 13 to alpha5beta1. Analysis of this inhibition showed that at high ligand concentrations, approximately 50% of the total integrin bound mAb 13 with >50-fold lower affinity than in the absence of ligand. The concentration of ligand required for half-maximal inhibition of antibody binding was independent of antibody concentration, suggesting that ligand acts as an allosteric inhibitor of mAb 13 binding. Hence, ligand and mAb 13 did not appear to compete directly for binding to alpha5beta1. The stimulatory anti-beta1 mAb 9EG7 was found to increase the maximum level of ligand binding approximately 2-fold, indicating that up to 50% of the total integrin could not bind ligand without 9EG7 stimulation. Analysis of mAb 13 binding in the presence of 9EG7 and ligand (i.e. maximal ligand occupancy) demonstrated that essentially all of the integrin bound mAb 13 with very low or zero affinity. Our results demonstrate that mAb 13 recognizes an epitope that is dramatically attenuated in the ligand-occupied form of alpha5beta1. Hence, since mAb 13 preferentially recognizes the unoccupied conformation of the integrin, the antibody may inhibit ligand binding by stabilizing the unoccupied state of alpha5beta1. In addition, we present evidence that the binding of mAb 13 to ligand-occupied alpha5beta1 may also induce a conformational change in the integrin, resulting in the displacement of ligand. PMID- 8702773 TI - Structure and function of the 10 S conformation of smooth muscle myosin. AB - Smooth myosin regulatory light chain (RLC) was exchanged with RLC labeled with benzophenone-4-iodoacetamide at Cys-108. Irradiation under conditions that favor the folded (10 S) conformation resulted in 10 S cross-linked myosin that could not unfold. Purified 10 S cross-linked myosin was cross-linked between the RLC of one head to light meromyosin between leucine 1554 and glutamate 1583, adjacent to a predicted noncoiled region, approximately 60 nm from the tip of the tail. At high ionic strength without actin, product release from one-half of the heads was slow (like 10 S) whereas the other half were activated. This suggests that tail binding to the RLC carboxyl-terminal domain stabilizes ionic interactions important to slow nucleotide release. With actin, product release from both (un)phosphorylated 10 S cross-linked myosin was from one slow population similar to unphosphorylated filaments. 10 S cross-linked myosin weakly bound actin (dissociation constant > 500 microM) and did not move actin in vitro. Single headed myosin did not fold or trap nucleotide. These and other data suggest that "trapping" occurs only with both heads and the tail binds to a newly formed site, which includes the RLC carboxyl-terminal domain, once trapping has occurred. PMID- 8702774 TI - Selective binding of FKBP12.6 by the cardiac ryanodine receptor. AB - The calcium release channels (CRC)/ryanodine receptors of skeletal (Sk) and cardiac (C) muscle sarcoplasmic reticulum (SR) are hetero-oligomeric complexes with the structural formulas (ryanodine recepter (RyR)1 protomer)4(FKBP12)4 and (RyR2 protomer)4(FKBP12.6)4, respectively, where FKBP12 and FKBP12.6 are isoforms of the 12-kDa receptor for the immunosuppressant drug FK506. The sequence similarity between the RyR protomers and FKBP12 isoforms is 63 and 85%, respectively. Using 35S-labeled FKBP12 and 35S-labeled FKBP12.6 as probes to study the interaction with CRC, we find that: 1) analogous to its action in skeletal muscle sarcoplasmic reticulum (SkMSR), FK506 (or analog FK590) dissociates FKBP12.6 from CSR; 2) both FKBP isoforms bind to FKBP-stripped SkMSR and exchange with endogenously bound FKBP12 of SkMSR; and 3) by contrast, only FKBP12. 6 exchanges with endogenously bound FKBP12.6 or rebinds to FKBP-stripped CSR. This selective binding appears to explain why the cardiac CRC is isolated as a complex with FKBP12.6, whereas the skeletal muscle CRC is isolated as a complex with FKBP12, although only FKBP12 is detectable in the myoplasm of both muscle types. Also, in contrast to the activation of the channel by removal of FKBP from skeletal muscle, no activation is detected in CRC activity in FKBP-stripped CSR. This differential action of FKBP may reflect a fundamental difference in the modulation of excitation-contraction coupling in heart versus skeletal muscle. PMID- 8702775 TI - A hydrophobic region within the adenovirus E1B 19 kDa protein is necessary for the transient inhibition of NF-kappaB activated by different stimuli. AB - The early transcribed adenovirus proteins E1A and E1B display a variety of functions in the transformation of primary rodent cells and the regulation of apoptosis and transcription. We have recently shown recently that the E1B 19 kDa protein from Adenovirus 5 (Ad 5) can functionally antagonize the stimulatory effect of E1A 13S on the human transcription factor NF-kappaB. Here we show that expression of E1B 19 kDa negatively interfered with the activation of NF-kappaB by different stimuli, such as the E1A 13S protein, and treatment with phorbol ester and tumor necrosis factor alpha. This suggests that E1B 19 kDa acts on a common upstream signaling event. Band shift experiments showed that expression of E1B 19 kDa impaired the generation of the nuclear, DNA-binding form of NF-kappaB. Domain mapping experiments employing various E1B 19 kDa mutants revealed the necessity of a hydrophobic Bcl-2 homology region between amino acids 90 and 96 for NF-kappaB inhibition. Co-transfection experiments showed that the inhibitory effect of E1B 19 kDa on E1A 13S-activated NF-kappaB transcription was gradually lost in the course of time. Thus the continuous stimulatory action of E1A 13S can finally override the antagonistic effects of E1B 19 kDa on NF-kappaB activity. In contrast to E1B 19 kDa, expression of the E1B 55 kDa protein did not result in a de novo activation of NF-kappaB, but co-stimulated the transcriptional potential of activated NF-kappaB. PMID- 8702776 TI - Identification of a novel GTPase, the inducibly expressed GTPase, that accumulates in response to interferon gamma. AB - Interferon gamma is a pleiotropic cytokine that regulates many immune functions. We have identified a novel protein, inducibly expressed GTPase (IGTP), whose expression was regulated by interferon gamma in macrophages. In mouse RAW 264.7 macrophages, IGTP mRNA levels were almost undetectable but increased within 1 h of exposure to interferon gamma, peaked at very high levels within 3 h, and remained at high levels to at least 48 h; pretreatment of the cells with cycloheximide blocked the majority of mRNA accumulation. In the mouse, the mRNA was highly expressed in thymus, spleen, lung, and small intestine. Using interspecific backcross analysis, the Igtp gene was mapped to mouse chromosome 11. The IGTP cDNA encoded a putative polypeptide of Mr 48,507 and pI 7.79 that contained three consensus GTP binding motifs, GXXXXGK(S/T), DXXG, and NTKXD. Both IGTP that had been immunoprecipitated from RAW cells and a glutathione S transferase IGTP fusion protein were able to convert GTP to GDP in vitro. Subcellular protein fractionation and Western blotting localized IGTP to the cytosol of RAW cells. In addition, the protein was homologous to proteins encoded by three previously cloned cDNAs, IRG-47, TGTP/Mg21, and LRG-47, and thus may be representative of a new family of interferon gamma-regulated GTPases. PMID- 8702777 TI - Mapping Escherichia coli elongation factor Tu residues involved in binding of aminoacyl-tRNA. AB - Two residues of Escherichia coli elongation factor Tu involved in binding of aminoacyl-tRNA were identified and subjected to mutational analysis. Lys-89 and Asn-90 were each replaced by either Ala or Glu. The four single mutants were denoted K89A, K89E, N90A, and N90E, respectively. The mutants were characterized with respect to thermal and chemical stability, GTPase activity, tRNA affinity, and activity in an in vitro translation assay. Most conspicuously tRNA affinities were reduced for all mutants. The results verify our structural analysis of elongation factor Tu in complex with aminoacyl-tRNA, which suggested an important role of Lys-89 and Asn-90 in tRNA binding. Furthermore, our results indicate helix B to be an important target site for nucleotide exchange factor EF-Ts. Also the mutants His-66 to Ala and His-118 to either Ala or Glu were characterized in an in vitro translation assay. Their functional roles are discussed in relation to the structure of elongation factor Tu in complex with aminoacyl-tRNA. PMID- 8702778 TI - Glucocorticoid repression of the mouse gonadotropin-releasing hormone gene is mediated by promoter elements that are recognized by heteromeric complexes containing glucocorticoid receptor. AB - We have identified two regions of the mouse gonadotropin-releasing hormone (GnRH) promoter, one between -237 and -201 (distal element) and the other between -184 and -150 (proximal element), which are required for glucocorticoid repression in transiently transfected GT1-7 cells. These sequences show no similarity to known positive or negative glucocorticoid response elements (nGREs) and do not function when placed upstream of heterologous viral promoters. The glucocorticoid receptor (GR) does not bind directly to the distal or proximal promoter elements but may participate in glucocorticoid repression of GnRH gene transcription by virtue of its association within multiprotein complexes at these nGREs. Electrophoretic mobility shift assays with GT1-7 nuclear extract demonstrate the presence of GR containing protein complexes on GnRH nGREs. One protein that co-occupies the distal nGRE in vitro along with GR is the POU domain transcription factor Oct-1. Thus, the tethering of GR to the GnRH distal nGRE, by virtue of a direct or indirect association with DNA-bound Oct-1, could play a role in hormone-dependent transcriptional repression of the GnRH gene. In contrast, Oct-1 does not appear to be a component of the GR-containing protein complex that is bound to the proximal nGRE. PMID- 8702779 TI - Haloenol lactone is a new isozyme-selective and active site-directed inactivator of glutathione S-transferase. AB - A haloenol lactone derivative has been synthesized and found to be an isozyme selective and active site-directed inactivator of glutathione S-transferase (GST). Preincubation of the haloenol lactone (100 microM) with murine Alpha, Mu, or Pi GST isozyme (1.0 microM) at pH 6.5, 37 degrees C resulted in time-dependent loss of enzyme activity with highly selective inhibition of the Pi isozyme (t1/2, approximately 2 min). In a separate experiment, a 10-fold excess of the lactone was incubated with GST-Pi isozyme at 37 degrees C for 3 h, followed by dialysis against Nanopure water. GST activity lost upon incubation with the lactone could not be restored by exhaustive dialysis, and only 8% of enzyme activity for the modified GST remained relative to the control that was treated identically except the lactone was omitted from the incubation. Both control and modified GST were characterized using electrospray ionization mass spectrometry. No native GST (23,478 Da) was observed in the spectrum of modified GST. Instead, protein incubated with the lactone exhibited an increase in molecular mass of 230 Da relative to control GST. The lactone (100 microM) was incubated with GST Pi isozyme (1.0 microM) in the presence of the competitive inhibitor S hexylglutathione (10 microM), which suppressed time-dependent inhibition of GST by the lactone. The results suggest that this haloenol lactone is an irreversible and active site-directed inhibitor of GST that appears to inhibit the enzyme through two consecutive steps of nucleophilic attack. PMID- 8702780 TI - Isolation and characterization of the hyperthermostable serine protease, pyrolysin, and its gene from the hyperthermophilic archaeon Pyrococcus furiosus. AB - The hyperthermostable serine protease pyrolysin from the hyperthermophilic archaeon Pyrococcus furiosus was purified from membrane fractions. Two proteolytically active fractions were obtained, designated high (HMW) and low (LMW) molecular weight pyrolysin, that showed immunological cross-reaction and identical NH2-terminal sequences in which the third residue could be glycosylated. The HMW pyrolysin showed a subunit mass of 150 kDa after acid denaturation. Incubation of HMW pyrolysin at 95 degrees C resulted in the formation of LMW pyrolysin, probably as a consequence of COOH-terminal autoproteolysis. The 4194-base pair pls gene encoding pyrolysin was isolated and characterized, and its transcription initiation site was identified. The deduced pyrolysin sequence indicated a prepro-enzyme organization, with a 1249-residue mature protein composed of an NH2-terminal catalytic domain with considerable homology to subtilisin-like serine proteases and a COOH-terminal domain that contained most of the 32 possible N-glycosylation sites. The archaeal pyrolysin showed highest homology with eucaryal tripeptidyl peptidases II on the amino acid level but a different cleavage specificity as shown by its endopeptidase activity toward caseins, casein fragments including alphaS1-casein and synthetic peptides. PMID- 8702781 TI - Import of RNA into Leishmania mitochondria occurs through direct interaction with membrane-bound receptors. AB - Cytoplasmic tRNAs are imported into the kinetoplast mitochondrion of Leishmania, but the mechanism of import is unknown, particularly whether RNA is transferred as a ribonucleoprotein complex through the protein import pathway or by a distinct receptor-mediated mechanism. Using isolated mitochondria, it was shown that a small, importable RNA, which is structurally homologous to tRNA, binds rapidly, specifically, and with high affinity to the mitochondrial surface in the absence of soluble protein factors to form an import intermediate. Two classes of binding site of apparent Kd 0.3 and 10 n, respectively, were distinguished. tRNA from Leishmania, but not yeast, competitively inhibited the binding. Northwestern blot analysis revealed the presence of a 15-kDa RNA binding protein on the mitochondrial surface. Whereas receptor binding was resistant to heparin and KCl, internalization was sensitive to both reagents. These results are consistent with the presence of a direct mechanism of receptor-mediated RNA import on Leishmania mitochondria. PMID- 8702782 TI - Critical residues for ligand binding in an I domain-like structure of the integrin beta1 subunit. AB - Several integrin alpha subunits have an inserted sequence of about 200 residues (the I or A domain) that is critical for ligand interactions. The presence of an I domain-like structure within the integrin beta subunit has been proposed based on the similarity of the hydropathy profiles and the homology of sequences between the alpha and beta subunits. This study was designed to determine whether the region of the beta1 subunit that includes residues 101-335 has the characteristics of an I domain. We found novel critical residues for ligand binding (Ser-132, Asn-224, Asp-226, Glu-229, Asp-233, Asp-267, and Asp-295, in addition to the previously reported Asp-130) using site-directed mutagenesis. The critical residues for ligand binding are located in several of loop structures of the region (or in a potential loop between an alpha helix and a beta strand), which have been predicted using multiple secondary structure prediction methods. The data suggest that the beta subunit has multiple disrupted critical oxygenated residues for ligand binding similar to those found in the alpha I domain. PMID- 8702783 TI - Regulation of Na+/H+ exchanger gene expression. Role of a novel poly(dA.dT) element in regulation of the NHE1 promoter. AB - In this study we examine regulation of expression of the Na+/H+ exchanger promoter in L6 and NIH 3T3 cells. We have identified a highly conserved poly(dA dT)-rich region that appears to be important in regulation of expression of the NHE1 gene. Deletion or mutation of this region results in dramatic decreases in promoter activity in both L6 and NIH 3T3 cells. In addition, DNase I footprinting experiments demonstrated that this region is protected by nuclear extracts from both cell types, and gel mobility shift assays showed that a protein or proteins specifically binds to the poly(dA dT)-rich element. Using Southwestern blotting, we determined that a 33-kDa protein binds to the poly(dA dT)-containing region. Mutations that abolished protein binding to this element diminished activity of the promoter. Insertion of the poly(dA dT)-rich element into a plasmid containing the SV40 promoter demonstrated that this element can also enhance the activity of a foreign promoter. Together, the results we have presented here show that the poly(dA dT)-rich region is important in regulation of NHE1 expression in different cell types. PMID- 8702784 TI - DNA end joining by the Klenow fragment of DNA polymerase I. AB - DNA end joining is a type of illegitimate recombination characterized by the joining of two DNA ends that lack homology. Using oligonucleotides as substrate, we found that an exonuclease-free derivative of the Klenow fragment of Escherichia coli DNA polymerase I can mediate DNA end joining in vitro. DNA sequence analysis of product DNA indicated that overlap products were formed between direct repeat sequences at the termini of the oligonucleotides. Formation of recombinant products was dependent on the strandedness of the substrate DNA, and the rate of product formation was dependent on the size of the potential overlap. With one to three complementary bases available for pairing at the 3' termini, there was an absolute requirement that one of the oligonucleotides be double-stranded, whereas with four complementary bases, products were also formed in reactions with single-stranded oligonucleotides. When noncomplementary nucleotides were added to the terminus of one of the oligonucleotides, product formation was delayed but not blocked. These data indicate that a DNA polymerase can mediate DNA double strand break rejoining in the absence of other proteins. PMID- 8702785 TI - ESM-1 is a novel human endothelial cell-specific molecule expressed in lung and regulated by cytokines. AB - We here report the identification of a novel human endothelial cell-specific molecule (called ESM-1) cloned from a human umbilical vein endothelial cell (HUVEC) cDNA library. Constitutive ESM-1 gene expression (as demonstrated by Northern blot and reverse transcription-polymerase chain reaction analysis) was found in HUVECs but not in the other human cell lines tested. The cDNA sequence contains an open reading frame of 552 nucleotides and a 1398-nucleotide 3' untranslated region including several domains involved in mRNA instability and five putative polyadenylation consensus sequences. The deduced 184-amino acid sequence defines a cysteine-rich protein with a functional NH2-terminal hydrophobic signal sequence. Searches in several data bases confirmed the unique identity of this sequence. A rabbit immune serum raised against the 14-kDa COOH terminal peptide of ESM-1 immunoprecipitated a 20-kDa protein only in ESM-1 transfected COS cells. Immunoblotting and immunoprecipitation of HUVEC lysates revealed a specific 20-kDa band corresponding to ESM-1. In addition, constitutive ESM-1 gene expression was shown to be tissue-restricted to the human lung. Southern blot analysis suggests that a single gene encodes ESM-1. A time dependent up-regulation of ESM-1 mRNA was seen after addition of tumor necrosis factor alpha (TNFalpha) or interleukin (IL)-1beta but not with IL-4 or interferon gamma (IFNgamma) alone. In addition, when IFNgamma was combined with TNFalpha, IFNgamma inhibited the TNFalpha-induced increase of ESM-1 mRNA level. These data suggest that ESM-1 may have potent implications in the areas of vascular cell biology and human lung physiology. PMID- 8702786 TI - Tyrosine phosphorylation-dependent suppression of a voltage-gated K+ channel in T lymphocytes upon Fas stimulation. AB - Selective cell death plays a critical role in the development of the immune system and in the elimination of target cells expressing foreign antigens. Most of programmed cell death occurs by apoptosis. Apoptotic cell death of lymphocytes can be triggered by ligation of APO-1/Fas (CD95) antigen (Suda, T., and Nagata, S. (1994) J. Exp. Med. 179, 873-879; Nagata, S., and Golstein, P. (1995) Science 267, 1449-1456). We find that activation of Fas leads to the inhibition of the voltage-dependent n-type K+ channels (Kv1.3) studied by patch clamp technique in Jurkat T lymphocytes. Tyrosine kinases have been shown to be crucial in Fas induced cell death (Eischen, C. M., Dick, C. J., and Leibson, P. J. (1994) J. Immunol. 153, 1947-1954). The inhibition of the current is correlated with the tyrosine phosphorylation of immunoprecipitated and blotted K+ channel protein. We show, that the Src-like protein-tyrosine kinase inhibitor herbimycin A and the deficiency of the p56(lck) tyrosine kinase in mutant Jurkat cells abolished the channel inhibition and phosphorylation by anti-Fas antibody, while reconstitution of the p56(lck) kinase partly restored these effects of Fas receptor triggering. These results suggest a regulation of n-type K+ channels by tyrosine kinases upon Fas receptor triggering, which might be important for apoptosis. PMID- 8702787 TI - Kinetics of interaction of Rab5 and Rab7 with nucleotides and magnesium ions. AB - We describe here the kinetics of the interaction of GTP and GDP with the small GTP-binding proteins Rab5 and Rab7. It was possible to make use of the intrinsic fluorescence of these proteins, since Rab5 contains two and Rab7 three tryptophan residues, respectively. With both enzymes, there is a significant decrease in fluorescence on binding GTP and an increase on binding GDP. As with the small GTP binding protein Ha-Ras p21 and with EF-Tu, nucleotide binding occurs in at least two steps and is describable in terms of a relatively weak initial interaction followed by a highly irreversible isomerization of the protein-nucleotide complex, which results in a change in the fluorescence properties. Dissociation of GDP and GTP could be followed in a time-dependent manner using fluorescently labeled GDP (methylanthraniloyl GDP) as displacing agent and taking advantage of substantial fluorescent energy transfer from tryptophan to the nucleotide. Fluorescence techniques could also be used to quantitate the interaction of Mg2+ ions with the GTP and GDP forms of Rab7, and it was shown that the metal ion was bound approximately 1000-fold more strongly to the GTP than the GDP form. The rate of GTP cleavage by the two proteins differed by a factor of approximately 20 (2 x 10(-3)s-1 for Rab5 and 9 x 10(-4)s-1 for Rab7 at 37 degrees C). Both proteins showed significant discrimination against xanthosine 5'-O-diphosphate (Kd approximately 10(3)-fold higher than that of GDP) and dramatic discrimination against ADP or ATP (Kd approximately 10(6)-fold higher than that of GDP). The results demonstrate a high degree of mechanistic similarity between the Rab proteins and other GTP-binding proteins, which have been examined in detail, including Ha-Ras p21, Ran, and EF-Tu. PMID- 8702788 TI - The crystallographic structure of phytohemagglutinin-L. AB - The structure of phytohemagglutinin-L (PHA-L), a leucoagglutinating seed lectin from Phaseolus vulgaris, has been solved with molecular replacement using the coordinates of lentil lectin as model, and refined at a resolution of 2.8 A. The final R-factor of the structure is 20.0%. The quaternary structure of the PHA-L tetramer differs from the structures of the concanavalin A and peanut lectin tetramers, but resembles the structure of the soybean agglutinin tetramer. PHA-L consists of two canonical legume lectin dimers that pack together through the formation of a close contact between two beta-strands. Of the two covalently bound oligosaccharides per monomer, only one GlcNAc residue per monomer is visible in the electron density. In this article we describe the structure of PHA L, and we discuss the putative position of the high affinity adenine-binding site present in a number of legume lectins. A comparison with transthyretin, a protein that shows a remarkable resemblance to PHA-L, gives further ground to our proposal. PMID- 8702789 TI - Defect in multiple cell cycle checkpoints in ataxia-telangiectasia postirradiation. AB - The recent description of a novel gene (ATM) mutated in ataxia-telangiectasia (A T), with homologies to genes encoding proteins involved in both G1/S and G2/M checkpoint control, points to a common defect in cell cycle control in A-T operating through the cyclin-dependent kinases. In this report we demonstrate that cyclin-dependent kinases are resistant to inhibition by ionizing radiation exposure in A-T cells, and this appears to be due to insufficient induction of WAF1. Exposure of control lymphoblastoid cells to radiation during S phase and in G2 phase causes a rapid inhibition of cyclin A-Cdk2 and cyclin B-Cdc2 activities, respectively. Irradiation led to a 5-20-fold increase in Cdk-associated WAF1 in these cells, which accounts at least in part for the decrease in cyclin-dependent kinase activity. In contrast, radiation did not inhibit any of the cyclin dependent kinase activities in S phase or G2 phase in A-T cells at short times after irradiation nor was there any significant change in the level of Cdk associated WAF1 compared to unirradiated cells. These results are similar to those reported previously for the G1 checkpoint and provide additional evidence for the involvement of ATM at multiple points in cell cycle regulation. PMID- 8702790 TI - Interferon-alpha-dependent activation of Tyk2 requires phosphorylation of positive regulatory tyrosines by another kinase. AB - Tyk2 and JAK1, members of the Janus kinase (JAK) family of protein tyrosine kinases, are required for interferon-alpha/beta binding and signaling. Both enzymes are associated with the interferon-alpha/beta receptor, and upon ligand binding, they undergo tyrosine phosphorylation and catalytic activation in an interdependent manner. To identify residues involved in Tyk2 regulation and to understand the basis of the interdependence of Tyk2 and JAK1, six mutated versions of Tyk2 bearing single or multiple point mutations in the tyrosine kinase domain were studied in a cell line lacking endogenous Tyk2. The Y1054F/Y1055F substitutions in the putative activation loop prevented ligand dependent activation of Tyk2, without abolishing its catalytic potential. The K930R mutation in the ATP binding site generated a kinase-negative protein, which however, still became phosphorylated upon interferon-alpha treatment. The Y1054F/Y1055F substitutions in this kinase-negative Tyk2 abolished the induced phosphorylation. These results indicate that Tyk2 is activated by phosphorylation on Tyr-1054 and/or Tyr-1055 and that this phosphorylation requires another kinase, most likely JAK1. While the Tyk2 forms mutated on Tyr-1054 and Tyr-1055 or on Lys-930 allowed some inducible gene expression, the combination of the three point mutations totally abolished signaling. PMID- 8702791 TI - Human T-cell leukemia virus type I Tax protein transactivates RNA polymerase III promoter in vitro and in vivo. AB - Tax protein of the human T-cell lymphotropic virus type 1 (HTLV-I) is critical for viral replication and is a potent transcriptional activator of viral and cellular polymerase II (pol II) genes. We report here that Tax is able to transactivate a classical pol III promoter, VA-I. In cotransfection experiments, Tax is shown to increase transcription of the VA-I promoter approximately 25 fold. Moreover, Tax is able to activate VA-I transcription when added exogenously to an in vitro transcription reaction. Using Tax affinity column chromatography, we demonstrate that Tax is able to deplete a HeLa cell extract for components required for transcription of VA-I. The transcriptional activity of the Tax depleted extract can be restored by the 0.6 phosphocellulose fraction. Interestingly, a consensus binding site for cAMP-responsive element binding protein (CREB) is located upstream of the VA-I promoter, and deletion of this element results in the loss of Tax responsiveness. When this CREB binding site is replaced by a Gal-4 binding site, the VA-I promoter can be transactivated by a Gal4-Tax fusion protein. Taken together, these results suggest that Tax may activate pol III and pol II promoter through a similar mechanism involving the CREB activation pathway. It is also possible that Tax affects pol III transcription by direct interaction with a component of the pol III transcriptional machinery. PMID- 8702792 TI - Docosahexaenoic acid modulates the interactions of the interphotoreceptor retinoid-binding protein with 11-cis-retinal. AB - Rapid transport of retinoids across the interphotoreceptor matrix is a critical part of the visual cycle, since it serves to replenish bleached rhodopsin with its chromophore 11-cis-retinal. The transport of retinoids in the interphotoreceptor matrix is believed to be mediated by the interphotoreceptor retinoid-binding protein (IRBP), a protein that, in addition to possessing two retinoid-binding sites, associates in vivo with long chain fatty acids. Here, the interrelationships between binding of the two types of ligands to IRBP were studied. The composition of fatty acids associated with IRBP in bovine retina was determined, and it was found that polyunsaturated fatty acids constitute a significant fraction of those. It was further found that docosahexaenoic acid, but not palmitic acid, induced a rapid and specific release of 11-cis-retinal from one of the protein's retinoid-binding sites. Based on these results and on the additional observation that a steep concentration gradient of docosahexaenoic acid exists between photoreceptor and pigment epithelium cells, a model for the mechanism by which IRBP may target 11-cis-retinal to photoreceptor cells is proposed. PMID- 8702793 TI - Dependence of fibroblast migration on actin severing activity of gelsolin. AB - Gelsolin nucleates actin filament assembly, blocks the fast-exchanging ends of actin filaments, and severs filaments, processes that may play an important role in cell motility. We studied the relationship between cell migration, gelsolin content, and actin severing activity in human gingival fibroblasts. These cells were keratin negative and desmin negative but expressed vimentin and myosin II. Cells were separated by their ability to migrate in response to a chemoattractant stimulus. Northern analysis of mRNA, [35S]methionine incorporation into immunoprecipitated gelsolin, immunoblots of cell lysates, and quantitative confocal microscopy showed 1.4-2-fold higher levels of gelsolin in nonmigrant compared with migrant cells. Because the concentration of intracellular gelsolin did not appear to be a central determinant of cell migration, we assessed its requirement for motility. Cells that were electroinjected with a gelsolin antibody that inhibits actin severing by gelsolin in vitro showed a 72% reduction of the number of migrant cells compared with controls treated with an irrelevant antibody. Cells that were electroinjected with free gelsolin exhibited a 33% increase in migration compared with cells electroinjected with bovine serum albumin. Compared with nonmigrant cells, migrant cells contained abundant free gelsolin and exhibited gelsolin-dependent F-actin severing activity, which required Ca2+. Serum stimulation of cell migration required increases in [Ca2+]i because incubation with 3 microM 1,2-bis-(o-aminophenoxy)-ethane-N,N, N',N' tetraacetic acid tetra(acetoxymethyl)-ester blocked calcium flux and cell migration. Serum also stimulated the recruitment of gelsolin into the supernatants of lysates from migrant but not from nonmigrant cells. In fibroblasts, gelsolin concentration alone does not apparently determine migratory capacity. Instead, the Ca2+-dependent actin severing activity of free gelsolin appears to be a major determinant of cell migration. PMID- 8702794 TI - Differential metabolic effects of adenovirus-mediated glucokinase and hexokinase I overexpression in rat primary hepatocytes. AB - The first step of glucose metabolism is the phosphorylation of glucose, catalyzed by the hexokinase family of enzymes. To address the metabolic impact of increasing glucose phosphorylation capacity in liver, rat primary hepatocytes were treated with recombinant adenoviruses containing the cDNAs encoding either rat liver glucokinase (AdCMV-GKL) or rat hexokinase I (AdCMV-HKI). Maximal glucose phosphorylation in AdCMV-GKL- and AdCMV-HKI-treated hepatocytes was increased 7.1 +/- 1.2- and 6.3 +/- 0.8-fold, respectively, over hepatocytes treated with an adenovirus expressing beta-galactosidase. Glucose usage (measured with 3 and 20 m 2-[3H]glucose and 5-[3H]glucose) was significantly increased in AdCMV-GKL-treated cells preincubated in 1 or 25 mM glucose. Treatment of hepatocytes with AdCMV-HKI also caused enhanced glucose utilization, but the increases were smaller and were less apparent in cells preincubated in high (25 mM) glucose. AdCMV-GKL-treated hepatocytes incubated for 48 h in the presence of variable glucose concentrations had glycogen levels that were maximally 15.0 +/- 0. 6-fold greater than levels in corresponding control cells. AdCMV-HKI-treated hepatocytes incubated under similar conditions had unchanged glycogen levels relative to controls. In AdCMV-GKL-treated cells, lactate output was increased to a maximum of 3.0 +/- 0.4-fold (at 25 mM glucose), glucose oxidation was increased 3.5 +/- 0.3-fold, and triglyceride production was unchanged relative to untreated cells. Among these three parameters, only lactate production was increased in AdCMV-HKI-treated cells, and then only at low glucose concentrations. We conclude that overexpression of glucokinase has potent effects on glucose storage and utilization in hepatocytes and that these effects are not matched by overexpression of hexokinase I. PMID- 8702795 TI - SCO1 and SCO2 act as high copy suppressors of a mitochondrial copper recruitment defect in Saccharomyces cerevisiae. AB - C129/U1 is a respiratory defective mutant of Saccharomyces cerevisiae arrested in cytochrome oxidase assembly due to a mutation in COX17, a nuclear gene encoding a low molecular weight cytoplasmic protein proposed to function in mitochondrial copper recruitment. In the present study we show that the respiratory defect of C129/U1 is rescuable by two multicopy suppressors, SCO1 and SCO2. SCO1 was earlier reported to code for a mitochondrial inner membrane protein with an essential function in cytochrome oxidase assembly (Buchwald, P., Krummeck, G., and Rodel, G. (1991) Mol. Gen. Genet. 229, 413-420). SCO2 is a homologue of SCO1, whose product is also localized in the mitochondrial membrane but is not required for respiration. SCO1 also suppresses a cox17 null mutant, indicating that overexpression of Sco1p can compensate for the absence of Cox17p. In contrast, neither copper, COX17 on a multicopy plasmid, or a combination of the two is able to restore respiration in sco1 mutants. Rescue of cox17 mutants by Sco1p suggests that this mitochondrial protein plays a role either in mitochondrial copper transport or insertion of copper into the active site of cytochrome oxidase. Although SCO2 can also partially restore respiratory growth in the cox17 null mutant, rescue in this case requires addition of copper to the growth medium. SCO2 does not suppress a sco1 null mutant, although it is able to partially rescue a sco1 point mutant. We interpret the ability of SCO2 to restore respiration in cox17, but not in sco1 mutants, to indicate that Sco1p and Sco2p have overlapping but not identical functions. PMID- 8702796 TI - Binding of anionic phospholipids to retinal pigment epithelium may be mediated by the scavenger receptor CD36. AB - The specific recognition of negatively charged phospholipids in cell membranes has been suggested to play an important role in a variety of physiological and pathophysiological processes. Recent work (Rigotti, A., Acton, S. L., and Krieger, M. (1995) J. Biol. Chem. 270, 16221-16224) has described specific and tight binding of anionic phospholipids, such as phosphatidylserine (PS) and phosphatidylinositol (PI), to the class B scavenger receptors, CD36 and SR-B1. We have previously reported that CD36 is present on retinal pigment epithelium (RPE) and plays a role in the phagocytosis of photoreceptor outer segments (ROS), a function critical to the normal visual process (Ryeom, S. W., Sparrow, J. R., and Silverstein, R. L. (1996) J. Cell Sci. 109, 387-395). We now report that phospholipid liposomes PS and PI, but not phosphatidylethanolamine, bind specifically to RPE. Cross-competition experiments suggest that PS and PI recognize the same receptor on RPE, while immunoinhibition studies indicate that the receptor is CD36. RPE cells isolated from a mutant rat strain, the RPE of which does not express CD36 ( Sparrow, J. R., Ryeom, S. W. , Abumrad, N., Ibrahimi, A., and Silverstein, R. L. (1996) Exp. Eye Res., in press), did not bind PS or PI, further confirming the role of CD36. We also showed that purified ROS blocked binding and uptake of anionic phospholipid liposomes by RPE and that PS and PI liposomes blocked ROS uptake by RPE, suggesting that PS and PI on the ROS membrane may be the ligands on ROS recognized by CD36. This is the first demonstration that CD36-phospholipid interactions may play a role in normal physiology. PMID- 8702797 TI - Diverging signal transduction pathways activated by interleukin 8 (IL-8) and related chemokines in human neutrophils. IL-8 and Gro-alpha differentially stimulate calcium influx through IL-8 receptors A and B. AB - Interleukin 8 (IL-8) and Gro-alpha are members of the CXC branch of a family of cytokines recently designated the "chemokine" superfamily. Recent evidence indicates that, contrary to previously held beliefs, IL-8 and Gro-alpha may not be perceived equivalently by neutrophils. In this study, we have evaluated the effects of IL-8 and Gro-alpha on the rate of calcium influx in human neutrophils and in 293 cells transfected with type A or type B IL-8 receptors. Of these two chemokines, only Gro-alpha induced an influx of calcium in neutrophils as judged by the sensitivity of the mobilization of calcium to the extracellular calcium chelator EGTA and to the nonselective divalent cation channel inhibitor SK&F 96365, as well as by manganese quenching experiments. IL-8 was similarly without effect on the rate of Mn2+ influx in 293 cells transfected with IL-8 receptor A (IL-8RA) or IL-8RB. On the other hand, Gro-alpha induced an SK&F 96365-sensitive increase of the rate of Mn+2 influx in IL-8RB-, but not in IL-8RA-transfected 293 cells. These results indicate not only that neutrophils respond differently to IL 8 than they do to Gro-alpha but, furthermore, that the consequences of the binding of IL-8 and Gro-alpha to IL-8RB are distinct. PMID- 8702798 TI - The CXC chemokines growth-regulated oncogene (GRO) alpha, GRObeta, GROgamma, neutrophil-activating peptide-2, and epithelial cell-derived neutrophil activating peptide-78 are potent agonists for the type B, but not the type A, human interleukin-8 receptor. AB - Interleukin-8 (IL-8), growth-related oncogene (GRO) alpha, GRObeta, GROgamma, neutrophil-activating peptide-2 (NAP-2), epithelial cell-derived neutrophil activating peptide- 78 (ENA-78), and granulocyte chemoattractant protein-2 are potent neutrophil chemoattractants 40-90% identical in amino acid sequence that comprise a subgroup of human CXC chemokines defined by the conserved sequence motif glutamic acid-leucine-arginine (ELR). Two human chemotactic receptor subtypes for IL-8, named IL-8 receptors (IL8R) A and B, have been cloned. They are 78% identical in amino acid sequence, coexpressed in neutrophils, and distinguished by their different selectivities for GROalpha and NAP-2. Their selectivity for other ELR+ CXC chemokines has not been previously reported. By measuring calcium flux in human embryonic kidney 293 cells transfected with plasmids encoding IL8RA or IL8RB, we have now defined receptor selectivity for GRObeta, GROgamma, and ENA-78. The rank order of agonist potency, based on inspection of the mean effective concentration values (EC50), for IL8RB was GROgamma (1 nM) > IL-8 (4 nM) approximately GROalpha (5 nM) approximately GRObeta (4 nM) approximately NAP-2 (7 nM) > ENA-78 (11 nM), and for IL8RA was IL-8 (4 nM) >>> ENA-78 (40 nM) approximately NAP-2 (45 nM) > GROalpha (63 nM) approximately GROgamma (65 nM) >> GRObeta. The maximal response of IL8RA to IL-8 was at least 2 fold greater than the other five chemokines. All six agonists for IL8RB competed for high affinity 125I-IL-8, -GROalpha, -NAP-2, and -ENA-78 binding sites at IL8RB. GROalpha, GRObeta, GROgamma, NAP-2, and ENA-78 competed weakly for the high affinity IL-8 binding site at IL8RA. Thus, IL8RA and IL8RB are both highly selective for IL-8 and have similar sequences but differ dramatically in their selectivity for all other ELR+ CXC chemokines tested. These findings have important implications for developing novel neutrophil-specific anti-inflammatory drugs directed against the CXC chemokine signaling system. PMID- 8702799 TI - Identification of functional domains within the RAD1.RAD10 repair and recombination endonuclease of Saccharomyces cerevisiae. AB - Saccharomyces cerevisiae rad1 and rad10 mutants are unable to carry out nucleotide excision repair and are also defective in a mitotic intrachromosomal recombination pathway. The products of these genes are subunits of an endonuclease which recognizes DNA duplex/single-strand junctions and specifically cleaves the 3' single-strand extension at or near the junction. It has been suggested that such junctions arise as a consequence of DNA lesion processing during nucleotide excision repair and the processing of double-strand breaks during intrachromosomal recombination. In this study we show that the RAD1 RAD10 complex also cleaves a more complex junction structure consisting of a duplex with a protruding 3' single-strand branch that resembles putative recombination intermediates in the RAD1 RAD10-mediated single-strand annealing pathway of mitotic recombination. Using monoclonal antibodies, we have identified two regions of RAD1 that are required for the cleavage of duplex/single-strand junctions. These reagents also inhibit nucleotide excision repair in vitro, confirming the essential role of the RAD1 RAD10 endonuclease in this pathway. PMID- 8702800 TI - Concentration dependence and time course of the effects of glucose on adenine and guanine nucleotides in mouse pancreatic islets. AB - Changes in the ATP:ADP ratio in pancreatic B cells may participate in the regulation of insulin secretion by glucose. Here, we have investigated the possible role of guanine nucleotides. Mouse islets were incubated in a control medium (when K+-ATP channels are the major site of regulation) or in a high K+ medium (when glucose modulates the effectiveness of cytosolic Ca2+ on exocytosis). Glucose induced a concentration-dependent (0-20 m) increase in GTP and a decrease in GDP in both types of medium, thus causing a progressive rise of the GTP:GDP ratio. ATP and ADP levels were 4-5-fold higher but varied in a similar way as those of guanine nucleotides. Insulin secretion was inversely correlated with ADP and GDP levels and positively correlated with the ATP:ADP and GTP:GDP ratios between 6 and 20 m glucose in control medium and between 0 and 20 m glucose in high K+ medium. The increases in the GTP:GDP and ATP:ADP ratios induced by a rise of glucose were faster than the decreases induced by a fall in glucose, but the changes of both ratios were again parallel. In conclusion, glucose causes large, concentration-dependent changes in guanine as well as in adenine nucleotides in islet cells. This raises the possibility that both participate in the regulation of nutrient-induced insulin secretion. PMID- 8702801 TI - Ca2+ translocation across sarcoplasmic reticulum ATPase randomizes the two transported ions. AB - Cytoplasmic Ca2+ dissociation is sequential, and the Ca2+ ions bound to the nonphosphorylated ATPase are commonly represented as superimposed on each other, so that the superficial Ca2+ is freely exchangeable from the cytoplasm, whereas the deeper Ca2+ is not. Under conditions where ADP-sensitive phosphoenzyme accumulates (leaky vesicles, 5 degrees C, pH 8, 300 mM K+), luminal Ca2+ dissociation is sequential as well, so that the representation of two superimposed Ca2+ ions still holds on the phosphoenzyme, with the superficial Ca2+ facing the lumen freely exchangeable and the deeper Ca2+ blocked by the superficial Ca2+. Under the same conditions, we have investigated whether a prebuilt Ca2+ order is maintained during membrane translocation. Starting from a prebuilt order on the cytoplasmic side, we showed that the Ca2+ ions cannot be identified after translocation to the luminal side. The same result was obtained starting from a prebuilt order on the luminal side and following the luminal to cytoplasmic translocation. We conclude that the two Ca2+ ions are mixed during ATP-induced phosphorylation as well as during ADP-induced dephosphorylation. PMID- 8702802 TI - Deficiency in beta1,3-galactosyltransferase of a Leishmania major lipophosphoglycan mutant adversely influences the Leishmania-sand fly interaction. AB - To study the function of side chain oligosaccharides of the cell-surface lipophosphoglycan (LPG), mutagenized Leishmania major defective in side chain biosynthesis were negatively selected by agglutination with the monoclonal antibody WIC79.3, which recognizes the galactose-containing side chains of L. major LPG. One such mutant, called Spock, lacked the ability to bind significantly to midguts of the natural L. major vector, Phlebotomus papatasi, and to maintain infection in the sand fly after excretion of the digested bloodmeal. Biochemical characterization of Spock LPG revealed its structural similarity to the LPG of Leishmania donovani, a species whose inability to bind to and maintain infections in P. papatasi midguts has been strongly correlated with the expression of a surface LPG lacking galactose-terminated oligosaccharide side chains. An in vitro galactosyltransferase assay using wild-type or Spock membranes was used to determine that the defect in Spock LPG biosynthesis is a result of defective beta1,3-galactosyltransferase activity as opposed to a modification of LPG, which would prevent it from serving as a competent substrate for galactose addition. The results of these experiments show that Spock lacks the beta1, 3-galactosyltransferase for side chain addition and that the LPG side chains are required for L. major to bind to and to produce transmissible infection in P. papatasi. PMID- 8702803 TI - Identification and characterization of Ich-3, a member of the interleukin-1beta converting enzyme (ICE)/Ced-3 family and an upstream regulator of ICE. AB - We report here the isolation and characterization of a new member of the ice/ced 3 family of cell death genes, named ich-3. The predicted amino acid sequence of Ich-3 protein shares 54% identity with murine interleukin-1beta converting enzyme (ICE). Overexpression of ich-3 in Rat-1 and HeLa cells induces apoptosis, which can be inhibited by CrmA and Bcl-2. The mRNA and proteins of ich-3 are dramatically induced in vivo upon stimulation with lipopolysaccharide, an inducer of septic shock. The ich-3 gene product can be cleaved by cytotoxic T cells granule serine protease granzyme B, suggesting that Ich-3 may mediate apoptosis induced by granzyme B. Ich-3 does not process proIL-1beta directly but does promote proIL-1beta processing by ICE. These results suggest that Ich-3 may play a very important role in apoptosis and inflammatory responses and may be an upstream regulator of ICE. PMID- 8702804 TI - Paclitaxel activation of the GADD153 promoter through a cellular injury response element containing an essential Sp1 binding site. AB - The GADD153 promoter is transcriptionally activated by paclitaxel-induced injury. Promoter deletion from -786 to -85 base pairs relative to the start of transcription had no significant effect on activation, but deletion to the TATA box abolished it. Placement of the 39 bases from -74 to the TATA box (cellular injury response element, CIRE) upstream of the adenovirus E4 TATA box conferred paclitaxel inducibility. The only consensus sequence present in the CIRE is an Sp1 site; mutation of this site inhibited paclitaxel activation. Paclitaxel failed to activate a SV40-driven luciferase construct containing five Sp1 sequences, and Sp1 sites further upstream in the GADD153 promoter were not essential for activation. Pure Sp1 and nuclear extracts from uninjured and paclitaxel-injured cells protected the same region from -62 to -48 bases on the noncoding strand and -74 to -53 on the coding strand. Nuclear extracts shifted the CIRE to the same extent as purified Sp1 but had no effect on a CIRE with a mutated Sp1 site in gel shift assays. Immunodepletion of Sp1 abolished the shift; antibody to Sp1 produced a supershift. These data indicate that paclitaxel activates the GADD153 promoter through a constitutively occupied Sp1 site at -61 bases. PMID- 8702805 TI - Characterization of the reductase domain of rat neuronal nitric oxide synthase generated in the methylotrophic yeast Pichia pastoris. Calmodulin response is complete within the reductase domain itself. AB - Rat neuronal NO synthase (nNOS) is comprised of a flavin-containing reductase domain and a heme-containing oxygenase domain. Calmodulin binding to nNOS increases the rate of electron transfer from NADPH into its flavins, triggers electron transfer from flavins to the heme, activates NO synthesis, and increases reduction of artificial electron acceptors such as cytochrome c. To investigate what role the reductase domain plays in calmodulin's activation of these functions, we overexpressed a form of the nNOS reductase domain (amino acids 724 1429) in the yeast Pichia pastoris that for the first time exhibits a complete calmodulin response. The reductase domain was purified by 2',5'-ADP affinity chromatography yielding 25 mg of pure protein per liter of culture. It contained 1 FAD and 0.8 FMN per molecule. Most of the protein as isolated contained an air stable flavin semiquinone radical that was sensitive to FeCN6 oxidation. Anaerobic titration of the FeCN6-oxidized reductase domain with NADPH indicated the flavin semiquinone re-formed after addition of 1-electron equivalent and the flavins could accept up to 3 electrons from NADPH. Calmodulin binding to the recombinant reductase protein increased its rate of NADPH-dependent flavin reduction and its rate of electron transfer to cytochrome c, FeCN6, or dichlorophenolindophenol to fully match the rate increases achieved when calmodulin bound to native full-length nNOS. Calmodulin's activation of the reductase protein was associated with an increase in domain tryptophan and flavin fluorescence. We conclude that many of calmodulin's actions on native nNOS can be fully accounted for through its interaction with the nNOS reductase domain itself. PMID- 8702806 TI - A new alternatively spliced exon between v9 and v10 provides a molecular basis for synthesis of soluble CD44. AB - The numerous isoforms of murine CD44 contain a common peptide region that is encoded by exons 1-5, 16-18, and 20 and variant regions derived from exons 6-15, usually referred to as v1-v10. We have obtained evidence for expression of an additional exon between v9 (or exon 14) and the exon previously termed v10 (or exon 15). Thus, we now number the variant exons as follows: v1-v9 (exons 6-14), v10 (exon 15), and v11 (exon 16); the remaining 3'-exons become exons 17-21 (newly numbered exons are underlined). The new exon, now termed exon v10, contains 93 base pairs and can be internally spliced; the 5'-region is termed v10a, and the 3'-region, v10b. Stop codons are positioned in v10a such that translated protein would be truncated prior to the transmembrane domain and secreted as a soluble protein. We have also found that the previously described v9 exon (now termed v9a), which is 90 base pairs in length, is actually the 5' region of a longer exon of 142 base pairs (the 3'-region is termed v9b) and thus arises by internal splicing of the longer exon. Using reverse transcription polymerase chain reaction, four different cDNAs for CD44 isoforms that use different combinations of the new exonic sequences have been found. The mRNAs containing the new exonic sequences are restricted in their expression; to date, we have demonstrated their presence in murine G8 myoblasts in culture and in embryonic muscle and cartilage tissues in vivo. Of these new isoforms, the predominant, full-length amplified product is encoded by exons 1-5, exon 13 (v8), the 5'-part of exon 14 (v9a), exon 15 (v10), exon 16 (v11), exons 17-19, and exon 21. When COS-7 cells were transfected with v10-containing cDNA constructs, the cells secreted low molecular weight soluble CD44 into their medium. Thus, the stop codons within the new exon v10 provide a molecular basis for de novo synthesis of soluble CD44 isoforms. PMID- 8702808 TI - Identification of the major urinary metabolite of the F2-isoprostane 8-iso prostaglandin F2alpha in humans. AB - F2-isoprostanes are prostaglandin-like products of nonenzymatic lipid peroxidation. Measurement of levels of endogenous unmetabolized F2-isoprostanes has proven to be a valuable approach to assess oxidative stress in vivo. However, measurement of levels of urinary metabolites of F2-isoprostanes in timed urine collections offers an advantage over measuring unmetabolized F2-isoprostanes, e. g. in a plasma sample, in that it can provide an integrated index of isoprostane production over time. Therefore, we sought to identify the major urinary metabolite in humans of one of the more abundant F2-isoprostanes produced, 8-iso prostaglandin F2alpha (8-iso-PGF2alpha). 20 microCi of tritiated 8-iso-PGF2alpha was infused over 1 h into a male volunteer. 75% of the infused radioactivity was excreted into the urine during the following 4.5 h and was combined with urine collected for 4 h from a rhesus monkey following infusion of 500 microg of unlabeled 8-iso-PGF2alpha. Urinary metabolites were isolated and purified by adsorption chromatography and high pressure liquid chromatography. The major urinary metabolite, representing 29% of the total extractable recovered radioactivity in the urine, was structurally identified by gas chromatography and mass spectrometry as 2,3-dinor-5, 6-dihydro-8-iso-prostaglandin F2alpha. The identification of 2, 3-dinor-5,6-dihydro-prostaglandin F2alpha as the major urinary metabolite of 8-iso-prostaglandin F2alpha provides the basis for the development of methods of assay for its quantification as a means to obtain an integrated assessment of oxidative stress status in humans. PMID- 8702807 TI - Cyclin D1 expression is regulated positively by the p42/p44MAPK and negatively by the p38/HOGMAPK pathway. AB - We have previously shown that the persistent activation of p42/p44(MAPK) is required to pass the G1 restriction point in fibroblasts (Pages, G., Lenormand, P., L'Allemain, G., Chambard, J. C., Meloche, S., and Pouyssegur, J. (1993) Proc. Natl. Acad. Sci. U. S. A. 90, 8319-8323) and postulated that MAPKs control the activation of G1 cyclin-dependent complexes. We examined the mitogen-dependent induction of cyclin D1 expression, one of the earliest cell cycle-related events to occur during the G0/G1 to S-phase transition, as a potential target of MAPK regulation. Effects exerted either by the p42/p44(MAPK) or the p38/HOGMAPK cascade on the regulation of cyclin D1 promoter activity or cyclin D1 expression were compared in CCL39 cells, using a co-transfection procedure. We found that inhibition of the p42/p44(MAPK) signaling by expression of dominant-negative forms of either mitogen-activated protein kinase kinase 1 (MKK1) or p44(MAPK), or by expression of the MAP kinase phosphatase, MKP-1, strongly inhibited expression of a reporter gene driven by the human cyclin D1 promoter as well as the endogenous cyclin D1 protein. Conversely, activation of this signaling pathway by expression of a constitutively active MKK1 mutant dramatically increased cyclin D1 promoter activity and cyclin D1 protein expression, in a growth factor independent manner. Moreover, the use of a CCL39-derived cell line that stably expresses an inducible chimera of the estrogen receptor fused to a constitutively active Raf-1 mutant (DeltaRaf-1:ER) revealed that in absence of growth factors, activation of the Raf > MKK1 > p42/p44MAPK cascade is sufficient to fully induce cyclin D1. In marked contrast, the p38(MAPK) cascade showed an opposite effect on the regulation of cyclin D1 expression. In cells co-expressing high levels of the p38(MAPK) kinase (MKK3) together with the p38(MAPK), a significant inhibition of mitogen-induced cyclin D1 expression was observed. Furthermore, inhibition of p38(MAPK) activity with the specific inhibitor, SB203580, enhanced cyclin D1 transcription and protein level. Altogether, these results support the notion that MAPK cascades drive specific cell cycle responses to extracellular stimuli, at least in part, through the modulation of cyclin D1 expression and associated cdk activities. PMID- 8702809 TI - Mechanisms of opsin activation. AB - Rhodopsin is constrained in an inactive conformation by interactions with 11-cis retinal including formation of a protonated Schiff base with Lys296. Upon photoisomerization, major structural rearrangements that involve protonation of the active site Glu113 and cytoplasmic acidic residues, including Glu134, lead to the formation of the active form of the receptor, metarhodopsin II b, which decays to opsin. However, an activated receptor may be generated without illumination by addition of all-trans-retinal or its analogues to opsin, as measured in this study by the increased phosphorylation of opsin by rhodopsin kinase. The potency of stimulation depended on the chemical and isomeric nature of the analogues and the length of the polyene chain with all-trans-C17 aldehyde and all-trans-retinal being the most active and trans-C12 aldehyde being the least active. Certain cis-isomers, 11-cis-13-demethyl-retinal and 9-cis-C17 aldehyde, were also active. Most of the retinal analogues tested did not regenerate a spectrally identifiable pigment, and many were incapable of Schiff base formation (ketone, stable oximes, and Schiff base-derivatives of retinal). Thus, receptor activation resulted from formation of non-covalent complexes with opsin. pH titrations suggested that an equilibrium exists between partially active (protonated) and inactive (deprotonated) forms of opsin. These findings are consistent with a model in which protonation of one or more cytoplasmic carboxyl groups of opsin is essential for activity. Upon addition of retinoids, the partially active conformation of opsin is converted to a more active intermediate similar to metarhodopsin II b. The model provides an understanding of the structural requirements for opsin activation and an interpretation of the observed activities of natural and experimental opsin mutants. PMID- 8702810 TI - Morphology and toxicity of Abeta-(1-42) dimer derived from neuritic and vascular amyloid deposits of Alzheimer's disease. AB - In the course of analyzing the chemical composition of Alzheimer's disease neuritic and vascular amyloid, we have purified stable dimeric and trimeric components of Abeta peptides. These peptides (molecular mass 9.0 and 13.5 kDa) were separated by size exclusion chromatography in the presence of 80% formic acid or 5 guanidine thiocyanate, pH 7.4. The average ratio of monomers, dimers, and trimers was 55:30:15, respectively. Similar structures were produced over time upon incubation of synthetic Abeta-(1-42) at pH 7.4. The stability of these oligomeric forms was also demonstrated by Western blot and mass spectrometry. Atomic force microscopy and electron microscopy rotary shadowing revealed that the monomers polymerized into 8-10-nm filaments, whereas the dimers generated prolate ellipsoids measuring 3-4 nm in diameter. The pathogenic effects of the dimeric Abeta-(1-40/42) were tested in cultures of rat hippocampal neuron glia cells. Only in the presence of microglia did the dimer elicit neuronal killing. It is possible that these potentially pathogenic Abeta-(1-40/42) dimers and trimers from Alzheimer's disease amyloid represent the soluble oligomers of Abeta recently described in Alzheimer's disease brains (Kuo, Y.-M., Emmerling, M. R., Vigo-Pelfrey, C., Kasunic, T. C., Kirkpatrick, J. B., Murdoch, G. H., Ball, M. J., and Roher, A. E. (1996) J. Biol. Chem., 271, 4077-4081). PMID- 8702811 TI - A protease processing site is essential for prorenin sorting to the regulated secretory pathway. AB - Transfected mouse pituitary AtT-20 cells were used to examine the sorting of human prorenin to dense core secretory granules and the regulated secretory pathway. These cells secrete prorenin constitutively and sort a portion of the prorenin to secretory granules, where it is converted to active renin by proteolytic processing. Pulse-chase labeling of transfected AtT-20 cells demonstrated that regulated secretion of prorenin was prevented by: 1) the mutagenic deletion of the prosegment, 2) the premature proteolytic removal of the prosegment by a Golgi-resident processing protease, or 3) the mutation of the native cleavage site so as to prevent removal of the prosegment. In addition, expression of fusion proteins containing portions of the prorenin prosegment demonstrated that exposure of potential proteolytic cleavage sites was sufficient to confer cleavage-dependent regulated secretion of the corresponding protein. These data implicate the protease cleavage event in the regulated secretion of prorenin and are consistent with the involvement of a subclass of processing proteases in the sorting of certain proteins to secretory granules in AtT-20 cells. PMID- 8702812 TI - Molecular characterization and developmental expression of a retinoid- and fatty acid-binding glycoprotein from Drosophila. A putative lipophorin. AB - A detailed understanding of the mechanism of lipid transport in insects has been hampered by the inability to identify the proapolipophorin gene that encodes apolipophorins I and II, the principal protein components of lipophorin, the lipid transport vehicle. Here we provide the first molecular description of the Drosophila gene encoding a retinoid- and fatty acid-binding glycoprotein (RFABG) and present evidence that it is a member of the proapolipophorin gene family. The gene, localized to the chromosome 4 (102 F region), encodes a 3351-amino acid protein that could serve as the precursor for the approximately 70-kDa and >200 kDa polypeptides associated with RFABG. The N-terminal sequence of the approximately 70-kDa polypeptide and that predicted for the >200-kDa polypeptide showed high sequence similarity to blowfly apolipophorin II and apolipophorin I, respectively. The RFABG precursor contains a signal peptide and exhibits a multidomain mosaic protein structure, which is typical of extracellular proteins. It has structural domains similar to lipid-binding proteins, namely vitellogenins and apolipoprotein B. The protein also contains a domain similar to the D domain of von Willebrand factor and mucin. The gene is expressed in the Drosophila embryo during development in cells that make up the amnioserosa and fat bodies. Immunolocalizations using specific antibodies against RFABG reveal that the protein is initially dispersed through the embryonic amnioserosa sac and latter concentrated at skeletal muscle-epidermis apodemeal contact junctions during larval development. This novel gene may play an important role in the transport of lipids, including retinoids and fatty acids, in insects. PMID- 8702813 TI - Competition for a unique response element mediates retinoic acid inhibition of vitamin D3-stimulated transcription. AB - We have identified a novel steroid hormone response element in the avian beta3 integrin promoter. This sequence, comprising three hexameric direct repeat half sites separated by nine and three nucleotides binds vitamin D receptor (VDR) retinoid X receptor (RXR) and retinoic acid receptor (RAR)-RXR heterodimers. VDR RXR binds direct repeats separated by three base pairs, and RAR-RXR recognizes half-sites separated by nine bases, whereas the central half-site interacts with both heterodimers. Retinoic acid and 1, 25-dihydroxyvitamin D3 activate both a genomic fragment including the transcriptional start site and an oligonucleotide containing the three repeats, linked to a heterologous promoter. Co-addition of the steroids produces neither synergy nor an additive effect; rather the result equals that for retinoic acid alone. Scatchard analysis demonstrates that RAR-RXR has greater affinity than VDR-RXR for the composite element. Based on these findings we propose a model in which there is specific, polarity-defined binding of VDR-RXR and RAR-RXR to three half-sites, which form two overlapping steroid response elements, with the central half-site common to both. Our results identify a novel mechanism by which one steroid hormone can modulate the activity of a second, by competing for a shared half-site in a composite response element. PMID- 8702814 TI - Two conserved tyrosine residues in protein R1 participate in an intermolecular electron transfer in ribonucleotide reductase. AB - The enzyme ribonucleotide reductase consists of two nonidentical proteins, R1 and R2, which are each inactive alone. R1 contains the active site and R2 contains a stable tyrosyl radical essential for catalysis. The reduction of ribonucleotides is radical-based, and a long range electron transfer chain between the active site in R1 and the radical in R2 has been suggested. To find evidence for such an electron transfer chain in Escherichia coli ribonucleotide reductase, we converted two conserved tyrosines in R1 into phenylalanines by site-directed mutagenesis. The mutant proteins were shown to be enzymatically inactive. In addition, the mechanism-based inhibitor 2'-azido-2'-deoxy-CDP was incapable of scavenging the R2 radical, and no azido-CDP-derived radical intermediate was formed. We also show that the loss of enzymatic activity was not due to impaired R1-R2 complex formation or substrate binding. Based on these results, we predict that the two tyrosines, Tyr-730 and Tyr-731, are part of a hydrogen-bonded network that constitutes an electron transfer pathway in ribonucleotide reductase. It is demonstrated that there is no electron delocalization over these tyrosines in the resting wild-type complex. PMID- 8702815 TI - The carboxyl terminus of GLUT4 contains a serine-leucine-leucine sequence that functions as a potent internalization motif in Chinese hamster ovary cells. AB - To characterize the trafficking motifs contained in the carboxyl terminus of GLUT4, a chimera (GTCTR) was constructed in which the carboxyl-terminal 30 amino acids of GLUT4 were substituted for the amino-terminal cytoplasmic domain of the transferrin receptor (TR). The endocytic behavior of this chimera was characterized in Chinese hamster ovary cells. The GTCTR chimera had a more predominant intracellular distribution compared to the TR. Only 20% of the GTCTR chimera is on the surface at steady-state compared to 35% of the TR. The GTCTR chimera is internalized 50% more rapidly and recycled 20% more slowly than the TR. Acidification of the cytosol inhibited internalization of the GTCTR chimera, indicating that the chimera is internalized through clathrin-coated pits. Mutations of GTCTR were constructed in which a di-leucine sequence of the carboxyl domain of GLUT4 was mutated to a di-alanine sequence (GTCTR-AA) and serine residue 488, immediately preceding the di-leucine sequence, was mutated to either an alanine or aspartate residue. In each case, albeit to varying degrees, the substitutions shifted the distribution of the mutated GTCTR constructs toward the surface. The shift in the distribution of GTCTR-AA resulted from a 10-fold reduction in internalization, and the shift of serine 488 mutants resulted from a 3-fold reduction in the internalization rate compared to GTCTR. None of these mutations affected the recycling rate. These results demonstrate that the carboxyl terminus of GLUT4 contains a serine-leucine-leucine-based motif that, when expressed in non-insulin responsive cells, functions as a potent internalization motif which promotes more rapid internalization than does the native TR internalization motif. PMID- 8702816 TI - Structural and functional characterization of OmpF porin mutants selected for larger pore size. I. Crystallographic analysis. AB - OmpF porin is a nonspecific pore protein from the outer membrane of Escherichia coli. Previously, a set of mutants was selected that allow the passage of long maltodextrins that do not translocate through the wild-type pore. Here, we describe the crystal structures of four point mutants and one deletion mutant from this set; their functional characterization is reported in the accompanying paper (Saint, N., Lou, K.-L., Widmer, C., Luckey, M., Schirmer, T., Rosenbusch, J. P. (1996) J. Biol. Chem. 271, 20676-20680). All mutations have a local effect on the structure of the pore constriction and result in a larger pore cross section. Substitution of each of the three closely packed arginine residues at the pore constriction (Arg-42, Arg-82, and Arg-132) by shorter uncharged residues causes rearrangement of the adjacent basic residues. This demonstrates mutual stabilization of these residues in the wild-type porin. Deletion of six residues from the internal loop (Delta109-114) results in disorder of seven adjacent residues but does not alter the structure of the beta-barrel framework. Thus, the large hollow beta-barrel motif can be regarded as an autonomous structure. PMID- 8702817 TI - Structural and functional characterization of OmpF porin mutants selected for larger pore size. II. Functional characterization. AB - The effects on the channel characteristics of four single amino acid substitutions in OmpF porin and of a deletion mutant in the constriction loop L3 have been studied. These mutations are all located in the narrow section of the channel of the protein that forms pores across the outer membrane of Escherichia coli. The single channel conductance of the deletion mutant (Delta109-114) is decreased by one third, whereas the point mutations do not exhibit significant deviations from that of the wild-type protein. The mutants exhibit drastic changes in ion selectivities. In the wild-type protein, the critical threshold potential (Vc), above which channels close reversibly, exhibits a strong pH dependence, with a titration point of approximately pH 7.7, which is abolished in all mutants studied here. Diffusion of six monosaccharides is little affected in the point mutants, while four disaccharides are taken up at highly increased rates by the deletion mutant. The functional results, presented here, are correlated to the x-ray structures of the mutants (Lou, K.-L., Saint, N., Prilipov, A., Rummel, G., Benson, S.A., Rosenbusch, J.P., and Schirmer, T. (1996) J. Biol. Chem. 271, 20669-20675). In most, but not all, cases, the structural changes explain the functional alterations observed. PMID- 8702818 TI - In vivo assembly of overproduced DNA polymerase III. Overproduction, purification, and characterization of the alpha, alpha-epsilon, and alpha-epsilon theta subunits. AB - The genes for the polymerase core (alphaepsilontheta) of the DNA polymerase III holoenzyme map to widely separated loci on the Escherichia coli chromosome. To enable efficient overproduction and in vivo assembly of DNA polymerase III core, artificial operons containing the three structural genes, dnaE, dnaQ, and holE, were placed in an expression plasmid. The proteins alpha, alphaepsilon and alphaepsilontheta were overexpressed and assembled in E. coli and purified to homogeneity. The three purified polymerases had a similar specific activity of about 6.0 x 10(6) units/mg in a gap-filling assay. Kinetics studies showed that neither epsilon nor theta influenced the Km of alpha for deoxynucleotide triphosphate and only slightly decreased the Km of alpha for DNA, although epsilon was absolutely required for maximal DNA synthesis. The rate of DNA synthesis by alpha-reconstituted holoenzyme using tau complex was about 5-fold less than that of alphaepsilon or alphaepsilontheta-reconstituted holoenzyme as determined by a gel analysis. The processivity of alpha-reconstituted holoenzyme was very similar to that of alphaepsilontheta-reconstituted holoenzyme when tau complex was used as a clamp loader. PMID- 8702819 TI - Biotin tagging deletion analysis of domain limits involved in protein macromolecular interactions. Mapping the tau binding domain of the DNA polymerase III alpha subunit. AB - The tau subunit dimerizes DNA polymerase III via interaction with the alpha subunit, allowing DNA polymerase III holoenzyme to synthesize both leading and lagging strands simultaneously at the DNA replication fork. Here, we report a general method to map the limits of domains required for heterologous protein protein interactions using surface plasmon resonance. The method employs fusion of a short biotinylation sequence at either the NH2 or COOH terminus of the protein to be immobilized on streptavidin-derivatized biosensor chips. Inclusion of a hexahistidine sequence permits rapid purification and separation of the fusion protein from the endogenous Escherichia coli biotin carboxyl carrier protein. Ten deletions of the alpha subunit were constructed and purified by Ni2+ nitrilotriacetic acid chromatography and, when required, monomeric avidin chromatography. Each alpha deletion protein was captured by streptavidin immobilized on a Pharmacia Biosensor BIAcore chip, and the tau binding activity of each alpha deletion was analyzed using surface plasmon resonance. The tau subunit bound very tightly to a full-length amino-terminal fusion of the biotinylation sequence with alpha (KD approximately 70 pm). Four additional NH2 terminal alpha deletion proteins (60, 240, 360, and 542 residues deleted) retained strong binding activity to the tau subunit (KD = 0.19-0.39 nM), whereas deletion of 705 residues or more from the NH2 terminus of the alpha subunit abolished tau binding activity. Full-length alpha that contained a carboxyl terminal fusion with the biotinylation sequence bound tau strongly (KD = 0.37 nM). However, deletion of 48 amino acids from the COOH terminus totally eliminated tau binding. These results indicate that the COOH-terminal half of the alpha subunit is involved in tau interaction. PMID- 8702820 TI - Identification of the beta-binding domain of the alpha subunit of Escherichia coli polymerase III holoenzyme. AB - Rapid and processive DNA synthesis by Escherichia coli DNA polymerase III holoenzyme is achieved by the direct interaction between the alpha subunit of DNA polymerase III core and the beta sliding clamp (LaDuca, R. J., Crute, J. J., McHenry, C. S., and Bambara, R. A. (1986) J. Biol. Chem. 261, 7550-7557; Stukenberg, T. P., Studwell-Vaughan, P. S., and O'Donnell, M. (1991) J. Biol. Chem. 266, 11328-11334). In this study, we localized the beta-binding domain of alpha to a carboxyl-terminal region by quantifying the interaction of beta with a series of alpha deletion proteins. Purification and binding analysis was facilitated by insertion of hexahistidine and short biotinylation sequences on the deletion terminus of alpha. Interaction of beta with alpha deletion proteins was studied by gel filtration and surface plasmon resonance. alpha lacking 169 COOH-terminal residues still possessed beta-binding activity; whereas deletion of 342 amino acids from the COOH terminus abolished beta binding. Deletion of 542 amino acids from the NH2 terminus of the 1160 residue alpha subunit resulted in a protein that bound beta 10-20-fold more strongly than native alpha. Hence, portions of alpha between residues 542 and 991 are involved in beta binding. DNA binding to alpha apparently triggers an increased affinity for beta (Naktinis, V., Turner, J., and O'Donnell, M. (1996) Cell 84, 137-145). Our findings extend this observation by implicating the amino-terminal polymerase domain in inducing a low affinity taut conformation in the carboxyl-terminal beta-binding domain. Deletion of the polymerase domain (or, presumably, its occupancy by DNA) relaxes the COOH-terminal domain, permitting it to assume a conformation with high affinity for beta. PMID- 8702821 TI - Normal T cells express two T cell antigen receptor populations, one of which is linked to the cytoskeleton via zeta chain and displays a unique activation dependent phosphorylation pattern. AB - The TCR couples antigen recognition and the transmission of activation signals. We report the expression of two TCR populations on the surface of T lymphocytes, one of which is linked to the cytoskeleton via the zeta chain. We also demonstrate that assembly of the CD3 subunits with cytoskeleton-associated zeta is necessary for their maximal localization to the cytoskeleton. The potential significance of these two receptor forms is underscored by differences observed in non-activated T cells; while detergent-soluble phosphorylated zeta appears as a 21-kDa protein, phosphorylated cytoskeleton-associated zeta appears as a 16-kDa form. This dichotomous phosphorylation pattern is rigidly maintained following activation, although each of the receptor populations undergoes different activation-dependent modifications: 1) levels of soluble phosphorylated 21-kDa zeta are enhanced, while phosphorylated 16-kDa cytoskeleton-associated zeta exhibits little change; 2) soluble non-phosphorylated 16-kDa zeta translocates to the cytoskeleton; 3) activation-dependent ubiquitinated zeta forms localize to both fractions, albeit with different kinetics. We also show that the protein tyrosine kinase Lck undergoes activation-dependent modifications and translocates to the cytoskeleton. The phosphorylation profiles of the dichotomous TCR populations in both non-activated and activated lymphocytes suggest that each population could regulate distinct cellular functions, possibly by select intermolecular associations. PMID- 8702822 TI - Mitogen-activated protein kinase-independent pathways mediate the effects of nerve growth factor and cAMP on neuronal survival. AB - Components of the mitogen-activated protein kinase (MAP kinase) signaling pathway, including Ras, Raf, and MAP kinase, are necessary for nerve growth factor (NGF)-induced neurite outgrowth in PC12 cells. We have investigated the role of this pathway in promoting survival of primary sympathetic neurons that die when deprived of NGF. NGF caused rapid and sustained increases (approximately 4-fold) in the activities of the ERK-1 and ERK-2 isoforms of MAP kinase. PD 098059, an inhibitor of MAP kinase kinase activation, blocked the effects of NGF on both kinase isoforms. However, PD 098059 did not attenuate the effects of NGF on neuronal survival. In addition, MAP kinase activity was not increased by chlorophenylthio-cAMP, a cell-permeable analog of cAMP that supports neuronal survival in the absence of NGF. These findings indicate that activation of MAP kinase is not required for the actions of either cAMP or NGF on neuronal survival. PMID- 8702823 TI - Cloning and functional characterization of a novel rat organic anion transporter mediating basolateral uptake of methotrexate in the kidney. AB - We have cloned a cDNA coding for a novel member of organic anion transporter, designated OAT-K1, expressed specifically in the kidney of rats. The rat OAT-K1 cDNA (2788 base pairs) had an open reading frame encoding for a 669-amino acid protein (calculated molecular mass of 74 kDa) which shows 72% identity with the cloned rat liver organic anion transporter, oatp. Northern hybridization and reverse transcription-coupled polymerase chain reaction revealed that the rat OAT K1 messenger RNA transcript is expressed predominantly in the kidney. By use of stable LLC-PK1 cell monolayers transfected with the rat OAT-K1 cDNA, the transporter was suggested to mediate basolateral uptake of methotrexate, an anionic anticancer drug, but not taurocholate, p-aminohippurate, prostaglandin E2, and leukotriene C4. The methotrexate transport by rat OAT-K1 was unaffected by the presence of Na+ or Cl- gradient. The methotrexate accumulation by the OAT K1-expressing cells showed saturability with the apparent Km value of 1.0 microM. Folate, sulfobromophthalein, and 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS) inhibited the methotrexate accumulation markedly. These findings suggest that the rat OAT-K1 is localized in the basolateral membranes of renal tubules, where it mediates renal clearance of methotrexate from the blood. PMID- 8702824 TI - Nucleosome disruption by human SWI/SNF is maintained in the absence of continued ATP hydrolysis. AB - We have examined the requirement for ATP in human (h) SWI/SNF-mediated alteration of nucleosome structure and facilitation of transcription factor binding to nucleosomal DNA. hSWI/SNF-mediated nucleosome alteration requires hydrolysis of ATP or dATP. The alteration is stable upon removal of ATP from the reaction or upon inhibition of activity by excess ATPgammaS, indicating that continued ATP hydrolysis is not required to maintain the altered nucleosome structure. This stable alteration is sufficient to facilitate binding of a transcriptional activator protein; concurrent ATP hydrolysis was not required to facilitate binding. These data suggest sequential steps that can occur in the process by which transcription factors gain access to nucleosomal DNA. PMID- 8702825 TI - Recruitment of activated p56lck on endosomes of CD2-triggered T cells, colocalization with ZAP-70. AB - We have previously established that upon CD2 activation of T cells, p56(lck) showed a transient increase in its kinase activity and was partially internalized. Here we studied the possibility that p56(lck) could retain its kinase activity in the endosomes of CD2-triggered cells. T cells were fractionated on a sucrose gradient, and the endosomal fraction was isolated. In CD2-triggered cells, part of Lck was internalized and presented a maximal kinase activity in the endosome-enriched fraction after 5 min, decreasing thereafter. In the endosomal fraction of activated cells, four tyrosine-phosphorylated proteins of apparent molecular masses of 30, 40, 56, and 70 kDa were detected. We demonstrated that the protein tyrosine kinase ZAP-70 was recruited to the endosomal fraction upon CD2 stimulation with kinetics similar to that of p56(lck), suggesting that recruitment of protein tyrosine kinases to endosomal vesicles could promote specific transduction signals at the intracellular level. PMID- 8702826 TI - Functional characterization of the transcription silencer element located within the human Pi class glutathione S-transferase promoter. AB - We have previously demonstrated enhanced transcriptional activity of the human Pi class glutathione S-transferase (GSTP1) promoter in a multidrug-resistant derivative (VCREMS) of the human mammary carcinoma cell line, MCF7 (Moffat, G. J., McLaren, A. W., and Wolf, C. R. (1994) J. Biol. Chem. 269, 16397-16402). Furthermore, we have identified an essential sequence (C1; -70 to -59) within the GSTP1 promoter that bound a Jun-Fos heterodimer in VCREMS but not in MCF7 cells. These present studies have examined the negative regulatory element (-105 to 86), which acted to suppress GSTP1 transcription in MCF7 cells. Mutational analysis of this silencer element further defined the repressor binding site to be located between nucleotides -97 and -90. In vitro DNA binding assays suggested that the repressor exerted its action by causing displacement of the essential non-AP-1-like MCF7 C1 complex. However, the addition of MCF7 nuclear extract did not disrupt binding of the VCREMS Jun-Fos C1 complex to the GSTP1 promoter. Furthermore, upstream insertion of the GSTP1 silencer element failed to inhibit activity of a heterologous promoter in MCF7 cells. These results highlighted the cell and promoter specificity of the GSTP1 transcriptional repressor and implicated a functional requirement for contact between the repressor and C1 complex. In this regard, the introduction of half-helical turns between the silencer and the C1 element abrogated repressor activity, thus leading to the hypothesis that a direct interaction between the repressor and C1 complex was required to suppress GSTP1 transcription. Moreover, these findings suggest that cell-specific differences in the composition of the C1 nuclear complex may dictate repressor activity. PMID- 8702827 TI - Arginine 75 in the pseudosubstrate sequence of type Ibeta cGMP-dependent protein kinase is critical for autoinhibition, although autophosphorylated serine 63 is outside this sequence. AB - Autoinhibitory domains in many protein kinases include either a phosphorylatable substrate-like sequence or a pseudosubstrate sequence. This study shows that Ibeta cGMP-dependent protein kinase (cGK) autophosphorylates Ser-63, which is in an atypical cGK substrate sequence (-59AQKQAS-) that is amino-terminal to the pseudosubstrate motif (-74KRQAI-). cGMP increases the rate of autophosphorylation (approximately 0.8 phosphate/cGK monomer) approximately 3-fold. Autophosphorylation is an intramolecular process since it is independent of cGK concentration. cGMP activation of cGK enhances proteolysis within and near the pseudosubstrate site; treatment of dimeric cGK with three proteases produces three cGK monomers (approximately 67-70 kDa each). Their amino-terminal sequences are 75RQAISAEPT-, 76QAISAEPTAF-, and 86DIQDLSXV-, respectively. cGMP stimulates these kinases by 10-, 2.5-, and 1.4-fold, respectively, compared with a 10-fold effect on intact cGK. Increased basal activity accounts for the diminished stimulation. Thus, the primary autophosphorylation site of Ibeta cGK is well outside the pseudosubstrate site, but Arg-75 in the pseudosubstrate site is critical for autoinhibition. Autoinhibition also involves elements that are carboxyl-terminal to Arg-75. PMID- 8702828 TI - Autophosphorylation of type Ibeta cGMP-dependent protein kinase increases basal catalytic activity and enhances allosteric activation by cGMP or cAMP. AB - Autophosphorylation of purified bovine Ibeta isozyme of cGMP-dependent protein kinase (Ibeta cGK) in the presence of cGMP or cAMP increased basal kinase activity (-cGMP) as much as 4-fold and reduced the Ka for both cGMP and cAMP; maximum catalytic activity (+cGMP) was not altered. Autophosphorylation proceeded with at least two rate components. The faster rate correlated with phosphorylation of Ser-63. The slower rate, as well as the increase in basal kinase activity and decrease in Ka for cyclic nucleotides, correlated with phosphorylation of Ser-79. Autophosphorylation of either residue was an intramolecular reaction. Autophosphorylation of a proteolytically generated Ibeta cGK monomer lacking amino-terminal residues 1-64 increased basal activity (3 fold) and decreased Ka for cAMP (15-fold). This indicated that autophosphorylation of Ser-79 did not require dimeric cGK and that the phosphorylation of Ser-79 in the monomer was sufficient to alter enzymatic characteristics of Ibeta cGK. These studies suggested that increases in intracellular cGMP or cAMP could result in autophosphorylation of Ibeta cGK, which would increase basal kinase activity as well as the sensitivity of cGK to activation by cGMP or to cross-activation by cAMP. Autophosphorylation could also prolong the increased kinase activity after decline of the second messenger. PMID- 8702829 TI - Constitutive activation of c-Met in liver metastatic B16 melanoma cells depends on both substrate adhesion and cell density and is regulated by a cytosolic tyrosine phosphatase activity. AB - Serial selection in vivo for liver colonization of B16 murine melanoma cells consistently resulted in cell lines expressing elevated amounts of the hepatocyte growth factor/scatter factor receptor (c-Met), which is constitutively activated in the absence of its cognate ligand. In this paper we present evidence suggesting that c-Met constitutive activation in liver-specific B16 melanoma cells depends on both receptor concentration on the cell surface and a cytosolic tyrosine phosphatase activity. In fact, c-Met constitutive activation is suddenly lost upon detachment of the cells from the substrate and is dramatically decreased in adherent cells plated at low density. The loss of tyrosine phosphorylation of c-Met in suspension appears to depend, at least partly, on an increased cytosolic tyrosine phosphatase activity. Instead, lower activation of c Met at low density mostly results from a decrease in receptor concentration on the membrane. Moreover, we show that c-Met activation does not occur homogeneously on the surface of adherent cells. In fact, receptor concentration and activation appear to be higher on the ventral surface (adherent to the substrate) than on the apical surface. Upon detachment, compartmentalization is lost, leading to a decrease in average receptor density on the plasma membrane and hence to a lower activation. PMID- 8702830 TI - Bacteriophage T4 anaerobic ribonucleotide reductase contains a stable glycyl radical at position 580. AB - It has been recently recognized that the class III anaerobic ribonucleotide reductase requires the presence of a second activating gene product, NrdG. We have proposed that the role for NrdG involves the generation of an oxygen sensitive glycyl free radical within the NrdD enzyme. In this article we present the generation of such a glycyl free radical within the T4 NrdD subunit and its dependence upon the phage NrdG subunit. Initially, an overexpression system was created that allowed the joint production of T4 NrdD and T4 NrdG. With this system and in the presence of T4 NrdG, an oxygen-sensitive cleavage of NrdD was observed that mimicked the cleavage observed in phage infected Escherichia coli extracts. Under anaerobic conditions the presence of T4 NrdD with NrdG revealed a strong doublet EPR signal (g = 2.0039). Isotope labeling of the NrdD with [2H]glycine and [13C]glycine, respectively, confirmed the presence of a stabilized glycine radical. The unpaired electron is strongly coupled to C-2 in glycine and the doublet splitting originates from one of the alpha-protons. The glycine residue at position 580 was determined to be the radical containing residue through site-directed mutagenesis studies involving a G580A NrdD mutant. The glycyl radical generation was specific for the T4 NrdG, and the host E. coli NrdG was found to be unable to activate the phage reductase. Finally, anaerobic purification revealed the holoenzyme complex to contain iron, whereas the NrdD polypeptide was found to lack the metal. Our results suggest a tetrameric structure for the T4 anaerobic ribonucleotide reductase containing one homodimer each of NrdD and NrdG, with a single glycyl radical present. PMID- 8702831 TI - Kidney N-acetylgalactosamine (GalNAc)-1-phosphate kinase, a new pathway of GalNAc activation. AB - A new enzyme that phosphorylates GalNAc at position 1 to form GalNAc-alpha-1P was purified approximately 1275-fold from the cytosolic fraction of pig kidney, and the properties of the enzyme were determined. The kinase is quite specific for GalNAc as the phosphate acceptor and is inactive with GlcNAc, ManNAc, glucose, galactose, mannose, GalN, and GlcN. This enzyme is clearly separated from galactokinase by chromatography on phenyl-Sepharose. The GalNAc kinase has a pH optimum between 8.5 and 9.0 and requires a divalent cation in the order Mg2+ > Mn2+ > Co2+, with optimum Mg2+ concentration at approximately 5 mM. The enzyme was most active with ATP as the phosphate donor, but slight activity was observed with ITP, acetyl-P, and phosphoenolpyruvate. Enzyme activity was highest in porcine and human kidney and porcine liver, but was low in most other tissues. Cultured HT-29 cells also had high activity for this kinase. The purified enzyme fraction was incubated with azido-[32P]ATP, exposed to UV light, and run on SDS gels. A 50-kDa protein was labeled, and this labeling showed saturation kinetics with increasing amounts of the probe and was inhibited by unlabeled ATP. Although the most purified GalNAc kinase preparation still had two bands that labeled with ATP, maximum labeling of the 50-kDa protein, but not the 66-kDa band, was coincident with maximum GalNAc kinase activity on a column of DEAE-Cibacron blue. On Sephacryl S-300, the native enzyme has a molecular mass of 48-51 kDa, indicating that the active kinase is a monomer. The product of the reaction was characterized as GalNAc-alpha-1-P by various chemical procedures. PMID- 8702832 TI - Inhibition of HIV-1 production and selective degradation of viral RNA by an amphibian ribonuclease. AB - Ribonucleases appear to have physiologic roles in host defense against cancer, viruses, and other parasites. Previously it was shown that select ribonucleases added to cells concurrently with virions blocked human immunodeficiency virus, type I (HIV-1) infection of H9 cells. We now report that a ribonuclease homologous to RNase A, named onconase, inhibits virus replication in chronically HIV-1-infected human cells without killing the virally infected cell. Examining the mechanism of this inhibition shows that onconase enters the infected cells and degrades HIV-1 RNA without degrading ribosomal RNA or the three different cellular messenger RNAs analyzed. The homologous human pancreatic RNase lacks anti-viral activity. Comparing recombinant forms of onconase and a onconase-human RNase chimera shows that the N-terminal 9 amino acids and the pyroglutamyl residue of onconase are required for full anti-viral activity. Thus extracellular ribonucleases can enter cells, metabolize select RNAs, and inhibit HIV virion production within viable replicating cells. PMID- 8702833 TI - The serum protein alpha2-HS glycoprotein/fetuin inhibits apatite formation in vitro and in mineralizing calvaria cells. A possible role in mineralization and calcium homeostasis. AB - We present data suggesting a function of alpha2-HS glycoproteins/fetuins in serum and in mineralization, namely interference with calcium salt precipitation. Fetuins occur in high serum concentration during fetal life. They accumulate in bones and teeth as a major fraction of noncollagenous bone proteins. The expression pattern in fetal mice confirms that fetuin is predominantly made in the liver and is accumulated in the mineralized matrix of bones. We arrived at a hypothesis on the molecular basis of fetuin function in bones using primary rat calvaria osteoblast cultures and salt precipitation assays. Our results indicate that fetuins inhibit apatite formation both in cell culture and in the test tube. This inhibitory effect is mediated by acidic amino acids clustering in cystatin like domain D1. Fetuins account for roughly half of the capacity of serum to inhibit salt precipitation. We propose that fetuins inhibit phase separation in serum and modulate apatite formation during mineralization. PMID- 8702834 TI - Characterization of the O-glycosylation sites in the chorionic gonadotropin beta subunit in vivo using site-directed mutagenesis and gene transfer. AB - Human chorionic gonadotropin (CG) is a member of a family of glycoprotein hormones which are heterodimers containing two nonidentical subunits: a common alpha and a hormone-specific beta subunit. One of the distinguishing features of the CGbeta subunit is the presence of four serine acceptors clustered within the last 25 amino acids. We previously demonstrated that this carboxyl-terminal region is important for maintaining its biologic half-life, and when the sequence was genetically fused to either the common alpha or follitropin beta subunits, O glycosylation was observed. Because this carboxyl-terminal sequence is located at the end of the subunit, we considered this region a convenient in vivo model for studying O-linked glycosylation in domains containing multiple serine recognition sites. A CGbeta gene was engineered in which the N-linked sites were inactivated to eliminate background from those carbohydrate groups. Using this construct, we made a series of truncation and amino acid substitutions of acceptor serines, and these mutants were transfected into Chinese hamster ovary cells. O-Glycosylation was determined by [3H]glucosamine incorporation and glycanase sensitivity of the products on SDS-polyacrylamide gels. We show that the O-linked sites comprise independent repetitive regions in which each acceptor serine has a recognition signal bounded by the next carboxy acceptor serine within four to five amino acids. It is also apparent that recognition of one site is not dependent on the glycosylation of another acceptor. Amino acid mutations in the acceptor regions demonstrated the importance of proline as a necessary feature for O-linked recognition in the CGbeta sequence. PMID- 8702835 TI - Cyclic AMP inhibition of thrombin-induced growth in vascular smooth muscle cells correlates with decreased JNK1 activity and c-Jun expression. AB - Thrombin is a potent modulator of vascular tone and vascular smooth muscle cell (VSMC) mitogenesis. Early studies from other laboratories demonstrated that cyclic AMP (cAMP) antagonizes the mitogenic effects of platelet-derived growth factor and epidermal growth factor by inhibiting the extracellular signal regulated protein kinases (ERKs; p42, p44) group of mitogen-activated protein kinases (MAPKs) in several cell types. This report examines the role of ERKs and Jun N-terminal kinase 1 (JNK1) groups of mitogen-activated protein kinases in thrombin-induced DNA synthesis in VSMCs using agents such as forskolin and dibutyrylcyclic AMP that increase intracellular cAMP levels. Both agents significantly inhibited thrombin-stimulated DNA synthesis in VSMCs. These agents, however, had no effect on thrombin induction of ERKs activation and c-Fos expression, suggesting divergence of the latter two events from the growth signaling events of thrombin that are sensitive to inhibition by cAMP. Thrombin activated JNK1 and induced c-Jun expression in VSMCs in a time-dependent manner. In contrast to ERKs and c-Fos, thrombin-induced JNK1 activation and c-Jun expression were sensitive to inhibition by forskolin, suggesting an association of these events with thrombin-stimulated growth in these cells. Thrombin also increased AP-1 activity, and this response was significantly blunted by forskolin. Together, these results demonstrate a correlation between JNK1 activation and c-Jun expression by thrombin and their association with the mitogenic signaling events of thrombin in VSMCs. PMID- 8702836 TI - Palmitoylation of endothelin receptor A. Differential modulation of signal transduction activity by post-translational modification. AB - Post-translational modifications such as phosphorylation and palmitoylation play important roles for the function and regulation of receptors coupled to heterotrimeric guanyl nucleotide-binding proteins. Here we demonstrate that the human endothelin receptor A (ETA) incorporates [3H]palmitate. Mutation of a cluster of five cysteine residues present in the cytoplasmic tail of ETA into serine or alanine residues completely prevented palmitoylation of the receptor. The ligand binding affinity of the non-palmitoylated ETA mutants was essentially unchanged as compared to the palmitoylated wild type ETA suggesting that the replacement of the cysteine residues did not alter the overall structure of the receptor. Furthermore, the ligand-induced stimulation of adenylyl cyclase by the mutant ETA was unaffected by the mutation. In contrast, the mutated non palmitoylated receptors but not the wild type receptor failed to stimulate phosphatidylinositol hydrolysis by phospholipase C activation upon challenge by endothelin-1. Furthermore, the mutant receptors failed to stimulate the ligand induced transient increase in the cytoplasmic calcium seen with the wild type ETA. Endothelin-1 induced mitogenic stimuli via the wild type receptors but not through the mutated receptors suggesting an important role for phospholipase C in this signal transduction pathway. The differential regulation of distinct signal transduction pathways by post-translational modification suggests that palmitoylation of the ETA provides a novel mechanism of modulating ETA receptor activity. PMID- 8702837 TI - An NF-kappaB site in the 5'-untranslated leader region of the human immunodeficiency virus type 1 enhances the viral expression in response to NF kappaB-activating stimuli. AB - The 5'-untranslated leader region of human immunodeficiency virus, type 1 (HIV 1), includes a complex array of putative regulatory elements whose role in the viral expression is not completely understood. Here we demonstrate the presence of an NF-kappaB-responsive element in the trans-activation response (TAR) region of HIV-1 that confers the full induction of HIV-1 long terminal repeat (LTR) in response to NF-kappaB-activating stimuli, such as DNA alkylating agents, phorbol 12-myristate 13-acetate, and tumor necrosis factor-alpha. The TAR NF-kappaB site GGGAGCTCTC spans from positions +31 to +40 and cooperates with the NF-kappaB enhancer upstream of the TATA box in the NF-kappaB-mediated induction of HIV-1 LTR. The conclusion stems from the following observations: (i) deletion of the two NF-kappaB sites upstream of the TATA box reduces, but does not abolish, the HIV-1 LTR activation by NF-kappaB inducers; (ii) deletion or base pair substitutions of the TAR NF-kappaB site significantly reduce the HIV-1 LTR activation by NF-kappaB inducers; (iii) deletions of both the NF-kappaB sites upstream of the TATA box and the TAR NF-kappaB site abolish the activation of HIV 1 LTR in response to NF-kappaB inducers. Moreover, the p50 p65 NF-kappaB complex binds to the TAR NF-kappaB sequence and trans-activates the TAR NF-kappaB directed expression. The identification of an additional NF-kappaB site in the HIV-1 LTR points to the relevance of NF-kappaB factors in the HIV-1 life cycle. PMID- 8702838 TI - Elevated cyclic AMP inhibits NF-kappaB-mediated transcription in human monocytic cells and endothelial cells. AB - The NF-kappaB/Rel family of transcription factors regulates the inducible expression of many genes in activated human monocytes and endothelial cells. In this study, we examined the molecular mechanism by which agents that elevate intracellular cAMP inhibit the expression of the tumor necrosis factor alpha (TNFalpha), tissue factor, endothelial leukocyte adhesion molecule-1, and vascular cell adhesion molecule-1 genes. Both forskolin and dibutyryl cAMP, which elevate intracellular cAMP by independent mechanisms, inhibited TNFalpha and tissue factor expression at the level of transcription. Induction of NF-kappaB dependent gene expression in transiently transfected human monocytic THP-1 cells and human umbilical vein endothelial cells was inhibited by elevated cAMP and by overexpression of the catalytic subunit of protein kinase A (PKA). Elevated cAMP did not prevent nuclear translocation of p50/p65 and c-Rel/p65 heterodimers, decrease nuclear translocation of p65, or significantly modify TNFalpha-induced phosphorylation of p65. Functional studies demonstrated that transcriptional activation of a plasmid containing multimerized kappaB sites by p65 was inhibited by agents that elevate cAMP and by overexpression of the catalytic subunit of PKA. This study indicates that activation of PKA reduces the induction of a distinct set of genes in monocytes and endothelial cells by inhibiting NF-kappaB mediated transcription. PMID- 8702839 TI - Genomic organization and chromosomal assignment of the human beta1, 4-N acetylgalactosaminyltransferase gene. Identification of multiple transcription units. AB - The beta1,4-N-acetylgalactosaminyltransferase (beta1,4GalNAc-T) (EC) gene is expressed in normal brain tissues and in various malignant transformed cells, such as malignant melanoma, neuroblastoma, and adult T cell leukemia. To analyze the regulatory mechanisms of gene expression, we determined the genomic organization of the beta1, 4GalNAc-T gene. The gene consists of at least 11 exons and spans >8 kilobase pairs. The coding region is located in exons 2-11. To determine the transcription initiation sites, 5'-rapid amplification of cDNA ends analysis and ribonuclease protection assays were performed using RNA obtained from the human melanoma cell line SK-MEL-31. Consequently, we defined three transcription initiation sites and the alternative usage of three exons. Exons 1a and 1b partially overlap; the latter is part (3'-side) of the former and corresponds to the 5'-noncoding region of the cDNA clone previously isolated. The third transcript, exon 1c, corresponds to nucleotides -520 to -412 (position +1 = A of ATG of beta1,4GalNAc-T cDNA), which are considered to be in intron 1 based on the cloned cDNA sequence. Ribonuclease protection assays revealed the corresponding protection bands in samples of the gene-expressing cell lines. 5' Flanking regions of individual initiation sites showed promoter activity when analyzed by chloramphenicol acetyltransferase assay in SK-MEL-31 cells. The multiple transcription initiation sites and their promoters/enhancers identified here might be differentially involved in the cell type-specific expression of the beta1,4GalNAc-T gene. This gene was assigned to human chromosome 12q13.3 by means of fluorescence in situ hybridization. PMID- 8702840 TI - Cytoplasmic O-GlcNAc modification of the head domain and the KSP repeat motif of the neurofilament protein neurofilament-H. AB - Neurofilaments, the major intermediate filaments in large myelinated neurons, are essential for specifying proper axonal caliber. Mammalian neurofilaments are obligate heteropolymers assembled from three polypeptides, neurofilament (NF)-H, NF-M, and NF-L, each of which undergoes phosphorylation at multiple sites. NF-M and NF-L are known to be modified by O-linked N-acetylglucosamine (O-GlcNAc) (Dong, D. L.-Y., Xu, Z.-S., Chevrier, M. R., Cotter, R. J., Cleveland, D. W., and Hart, G. W. (1993) J. Biol. Chem. 268, 16679-16687). Here we further report that NF-H is extensively modified by O-GlcNAc at Thr53, Ser54, and Ser56 in the head domain and, somewhat surprisingly, at multiple sites within the Lys-Ser-Pro repeat motif in the tail domain, a region in assembled neurofilaments known to be nearly stoichiometrically phosphorylated on each of the approximately 50 KSP repeats. Beyond the earlier identified sites on NF-M and NF-L, O-GlcNAc sites on Thr19 and Ser34 of NF-M and Ser34 and Ser48 of NF-L are also determined here, all of which are localized in head domain sequences critical for filament assembly. The proximity of O-GlcNAc and phosphorylation sites in both head and tail domains of each subunit indicates that these modifications may influence one another and play a role in filament assembly and network formation. PMID- 8702841 TI - The ability of the inhibitory domain of the POU family transcription factor Oct-2 to interfere with promoter activation by different classes of activation domains is dependent upon the nature of the basal promoter elements. AB - The Oct-2 transcription factor contains an inhibitory domain which is able to repress transcription following DNA binding. Here we show that within the neuronally expressed Oct-2.5 form, the inhibitory domain can strongly inhibit activation by transcription factor activation domains which are either composed predominantly of acidic residues or contain the HOB motif, whereas it has a weaker effect or no effect on proline-rich activation domains and on a glutamine rich domain. In contrast, the isolated inhibitory domain of Oct-2 can efficiently repress all types of activation domains. This effect is observed however, only on TATA box-containing promoters and not on promoters containing an initiator motif. This widespread inhibition of different activation domains and its dependence on the nature of the basal promoter elements indicate that the inhibitory domain is likely to act by contacting a common downstream target of activation domains within the basal transcriptional complex bound at the TATA box rather than quenching specific activation domains by direct interaction. These effects are discussed in terms of the functional role of the inhibitory domain within Oct-2.5 and the mechanism by which it acts. PMID- 8702842 TI - The amino-terminal module of the C4b-binding protein beta-chain contains the protein S-binding site. AB - Human C4b-binding protein (C4BP) is composed of multiple alpha-chains associated with a single beta-chain. Each chain is composed of homologous, tandemly arranged repeats of so-called short consensus repeats (SCRs). We have previously shown that the three SCR modules of the beta-chain contain a high affinity binding site for anticoagulant vitamin K-dependent protein S. On the basis of experiments using synthetic peptides, residues 31-45 of the amino-terminal SCR (SCR-1) in the beta-chain were suggested to be involved in protein S binding, but it is not known whether SCR-1 contains the entire protein S-binding site. To address this question, two different truncated forms of the beta-chain (beta1,2 and beta2, 3) were expressed in a prokaryotic expression system. The beta1,2 construct (SCR-1 + SCR-2) contained the high affinity binding site for protein S in contrast to beta2,3 (SCR-2 + SCR-3), which did not bind protein S. Unfortunately, it was not possible to express SCR-1 alone in this system. To further elucidate whether the protein S-binding site is fully contained in SCR-1 or whether SCR-2 is also required, recombinant alpha/beta-chain chimeras were constructed. These chimeras were composed of alpha-chains with one, two, or three of the amino-terminal SCR modules replaced by the beta-chain counterpart and were expressed in a eukaryotic expression system. All recombinant variants were retained within the cells and could be extracted in biologically active forms. The three alpha/beta-chain chimeras bound protein S equally well, with a Ka of approximately 2.3 x 10(8) +/- 0.2 M-1 as compared with 2.1 x 10(8) +/- 0.3 M-1 for plasma-purified C4BP. These results show that the entire protein S-binding site on C4BP is contained within beta-chain SCR-1. PMID- 8702843 TI - Purification, gene cloning, and reconstitution of the heterotrimeric single stranded DNA-binding protein from Schizosaccharomyces pombe. AB - We have purified a single-stranded DNA-binding protein (SSB) from Schizosaccharomyces pombe (Sp) and have shown that it is composed of three subunits of 68, 30, and 12 kDa. The SpSSB supports T antigen-dependent unwinding of SV40 ori containing DNA, but is not functional in the SV40 in vitro replication reaction. All three genes that encode the SpSSB subunit have been isolated. The cloned cDNA of the ssb1(+), encoding the p68 subunit, contains 609 amino acids (68.3 kDa), while that of the ssb2(+), encoding the p30 subunit, contains a 279 amino acids (30.3 kDa). The genomic DNA clone of the p12 subunit gene (ssb3(+)) has 2 introns and an open reading frame of 104 amino acids (11.8 kDa). Significant homology is observed among the largest and middle subunits of eukaryotic SSBs, but there is poor homology among the smallest subunits. In addition, we have reconstituted the SpSSB complex by coexpression of all three subunits in Escherichia coli. The reconstituted complex is active in single stranded DNA binding and the T antigen-dependent unwinding of SV40 ori DNA. Finally, we observed a cell cycle-dependent phosphorylation pattern of the p30 subunit of SpSSB, which is similar to that observed for the human and Saccharomyces cerevisiae SSB. PMID- 8702844 TI - Identification of a negative glucocorticoid response element in the rat type 1 vasoactive intestinal polypeptide receptor gene. AB - Glucocorticoids play important roles in lung development and function by modulating the expression of a variety of genes. The type 1 vasoactive intestinal polypeptide (VIP) receptor gene is highly expressed in the lung where it mediates VIP physiological functions. In this study, the effect of glucocorticoid on VIP receptor gene expression was examined. Dexamethasone (100 n) suppresses endogenous VIP receptor mRNA expression in cultured lung cells. Transient transfection of lung cells with fusion constructs containing various portions of the VIP receptor 5'-flanking sequences linked to the luciferase reporter gene shows that 126 base pairs (bp) of the VIP receptor upstream sequences are sufficient to mediate transcriptional repression by glucocorticoid. DNase I footprinting demonstrates that purified glucocorticoid receptor (GR) binds to the VIP receptor promoter between -21 and -36 bp relative to the transcription start site. Point mutations within this binding site not only abolish GR binding and GR mediated transcriptional repression of the VIP receptor gene, but basal transcription is also reduced to background levels. Co-transfection of GR expression vector and the VIP receptor GR binding site linked to the thymidine kinase promoter and luciferase shows that this sequence is sufficient to confer glucocorticoid-mediated transcriptional repression to a heterologous promoter. These results indicate that the VIP receptor gene contains a negative glucocorticoid response element between -21 and -36 bp that may act to regulate both basal and glucocorticoid-mediated expressions of the VIP receptor gene in lung cells. PMID- 8702845 TI - The dopamine transporter carboxyl-terminal tail. Truncation/substitution mutants selectively confer high affinity dopamine uptake while attenuating recognition of the ligand binding domain. AB - In order to delineate structural motifs regulating substrate affinity and recognition for the human dopamine transporter (DAT), we assessed [3H]dopamine uptake kinetics and [3H]CFT binding characteristics of COS-7 cells transiently expressing mutant DATs in which the COOH terminus was truncated or substituted. Complete truncation of the carboxyl tail from Ser582 allowed for the expression of biphasic [3H]dopamine uptake kinetics displaying both a low capacity (Vmax approximately 0.4 pmol/10(5) cells/min) high affinity (Km approximately 300 nM) component and one exhibiting low affinity (Km approximately 15 microM] and high capacity (Vmax approximately 5 pmol/10(5)cells/min) with a concomitant 40% decrease in overall apparent Vmax relative to wild type (WT) DAT. Truncation of the last 22 amino acids or substitution of the DAT-COOH tail with sequences encoding the intracellular carboxyl-terminal of either dopamine D1 or D5 receptors produced results that were identical to those with the fully truncated DAT, suggesting that the induction of biphasic dopamine uptake kinetics is likely conferred by removal of DAT-specific sequence motifs distal to Pro597. The attenuation of WT transport activity, either by lowering levels of DAT expression or by pretreatment of cells with phorbol 12-myristate 13-acetate (1 microM), did not affect the kinetics of [3H]dopamine transport. The estimated affinity of dopamine (Ki approximately 180 nM) for all truncated/substituted DAT mutants was 10-fold lower than that of WT DAT (approximately 2000 nM) and appears selective for the endogenous substrate, since the estimated inhibitory constants for numerous putative substrates or uptake inhibitors were virtually identical to those obtained for WT DATs. In marked contrast, DAT truncation/substitution mutants displayed significantly reduced high affinity [3H]CFT binding interactions with estimated Ki values for dopamine and numerous other substrates and inhibitors tested from 10-100-fold lower than that observed for WT DAT. Moreover, co-expression of truncated and/or substituted DATs with WT transporter failed to reconstitute functional or pharmacological activities associated with both transporters. Instead, complete restoration of uniphasic low affinity [3H]dopamine uptake kinetics (Km approximately 2000 nM) and high affinity substrate and inhibitor [3H]CFT binding interactions attributable to WT DATs were evident. These data clearly suggest the functional independence and differential regulation of the dopamine translocation process from the characteristics exhibited by its ligand binding domain. The lack of functional phenotypic expression of mutant DAT activities in cells co-expressing WT transporter is consistent with the contention that native DATs may exist as multisubunit complexes, the formation and maintenance of which is dependent upon sequences encoded within the carboxyl tail. PMID- 8702846 TI - Degradation and endoplasmic reticulum retention of unassembled alpha- and beta subunits of Na,K-ATPase correlate with interaction of BiP. AB - Assembly of alpha- and beta-subunits in the endoplasmic reticulum is a prerequisite for the structural and functional maturation of oligomeric P-type ATPases. In Xenopus oocytes, overexpressed, unassembled alpha- and beta-subunits of Xenopus Na,K-ATPase are retained in the endoplasmic reticulum (ER) and are degraded with different kinetics, while unassembled beta-subunits of gastric H, K ATPase leave the ER. In this study, we have investigated the role of the immunoglobulin-binding protein, BiP, in the folding, assembly, and ER retention of ATPase subunits. We determined the primary sequence of Xenopus BiP and used polyclonal antibodies to examine the interaction with BiP of various wild type and mutant alpha- and beta-subunits overexpressed in Xenopus oocytes. Our results show that ER-retained, unassembled Na,K-ATPase beta-subunits, but not transport competent H,K-ATPase beta-subunits, efficiently associate with BiP until assembly with alpha-subunits occurs. Furthermore, the kinetics of BiP interaction with unassembled wild type and with mutant Na,K-ATPase beta-subunits parallels their respective stability against cellular degradation. Finally, alpha-subunits that are overexpressed in oocytes and are rapidly degraded and endogenous oocyte alpha subunits that are stably expressed as individual assembly-competent proteins also interact with oocyte or exogenous BiP, and the interaction time correlates with the protein's stability. These data demonstrate for the first time that BiP might be involved in a long term maturation arrest and/or in the ER quality control of a multimembrane-spanning protein and lend support for a universal chaperone function of BiP. PMID- 8702847 TI - Purification of a 3beta-hydroxy-delta5-C27-steroid dehydrogenase from pig liver microsomes active in major and alternative pathways of bile acid biosynthesis. AB - A 3beta-hydroxy-Delta5-C27-steroid dehydrogenase active in bile acid biosynthesis was purified from pig liver microsomes by solubilization with sodium cholate and by chromatography on DEAE-Sepharose, aminohexyl-Sepharose, and blue Sepharose. The last step in the purification procedure was preparative isoelectric focusing in a Rotofor cell. The final enzyme preparation showed only one protein band upon SDS-polyacrylamide gel electrophoresis. The isoelectric point was estimated to about 7.0 and the apparent Mr was 36,000. The purified enzyme catalyzed the conversion of 7alpha-hydroxycholesterol, 7alpha,25-dihydroxycholesterol, 7alpha, 27-dihydroxycholesterol, and 3beta,7alpha-dihydroxy-5-cholestenoic acid into the corresponding 3-oxo-Delta4 compounds. The enzyme was inactive with C19 and C21 steroids as substrates. The enzyme was also inactive with C27 steroids having the 7-hydroxy group in beta- instead of alpha-position. The Km was found to be 0.30 and 0.32 microM with 7alpha-hydroxycholesterol and 7alpha, 27 dihydroxycholesterol as substrates, respectively. NAD+ was the preferred cofactor. A monoclonal antibody raised against the 3beta-hydroxy-delta5-C27 steroid dehydrogenase was prepared. After coupling to Sepharose, the antibody was able to bind the dehydrogenase and to decrease the conversion of 7alpha hydroxycholesterol into 7alpha-hydroxy-4-cholest-3-one by more than 90%. The N terminal amino acid sequence was determined and found to be similar but not identical with those of known 3beta-hydroxy-Delta5-steroid dehydrogenases active in steroid hormone biosynthesis. Thus, the purified enzyme active toward C27 steroids in bile acid biosynthesis appears to represent a novel type of 3beta hydroxy-delta5-steroid dehydrogenase. PMID- 8702848 TI - Purification and properties of a cytosolic V1-ATPase. AB - The native V1 complex of the tobacco hornworm vacuolar type ATPase (V-ATPase) was purified from cytosolic extracts of molting larval midgut. It consisted of the established V-ATPase subunits A, B, and E along with the 14-kDa subunit F and the novel 13-kDa subunit G. The final amount of purified V1 complex made up an unexpectedly high 2% of the total cytosolic protein, with a yield of approximately 0.4 mg/g of tissue. An equally high amount of cytosolic V1 complex was obtained from starving intermolt larvae. By contrast, the cytosolic V1 pool was reduced drastically in feeding intermolt larvae or in larvae that had been refed after starvation. The activity of the membrane-bound V-ATPase holoenzyme was inversely related to the size of the cytosolic V1 pool, suggesting that the insect plasma membrane V-ATPase is regulated by reversible disassembly of the V1 complex as a function of the feeding condition of the larvae. Like F1-ATPases, the purified V1 complex exhibited Ca2+-dependent ATPase activity and, in the presence of 25% methanol, exhibited Mg2+-dependent ATPase activity. Therefore, we designate the native V1 complex, V1-ATPase. Both enzyme activities were completely inhibited by micromolar N-ethylmaleimide. In contrast to the Ca2+ dependent V1-ATPase activity, the Mg2+/methanol-dependent V1-ATPase activity did not decrease with the incubation time and thus was not inhibited by ADP. Methanol appears to induce a conformational change of the V1 complex, leading to enzymatic properties of the V1-ATPase that are similar to those of the membrane-bound V ATPase holoenzyme. This is the first time that a native and enzymatically active V1 complex has been purified from the cytosol. PMID- 8702849 TI - Characterization of a new isoform of the NFAT (nuclear factor of activated T cells) gene family member NFATc. AB - The cyclosporin A (CsA)/FK506-sensitive nuclear factor of activated T cells (NFAT) plays a key role in the inducible expression of cytokine genes in T cells. Although NFAT has been recently shown to be inducible in several non-T immune cells, the NFAT gene family members characterized to date have been isolated only from T cells. To further characterize NFAT function in human B cells and to demonstrate cytokine gene specificity of NFAT proteins, we report here the isolation and characterization of a cDNA clone from the Raji B cell line. The cDNA clone encodes a new isoform, NFATc.beta, of the NFAT gene family member NFATc (designated here NFATc.alpha). The amino acid sequence of NFATc.beta differs from that of NFATc. alpha in the first NH2-terminal 29 residues and contains an additional region of 142 residues at the COOH terminus. Northern analysis using a probe encompassing a common region of both isoforms showed two mRNA species of 2.7 and 4.5 kilobase pairs, while an NFATc.beta-specific probe detected only the 4.5-kilobase pair mRNA which was preferentially expressed in the spleen. Transient expression of NFATc.beta was capable of activating an interleukin-2 NFAT-driven reporter gene in stimulated Jurkat cells in a CsA sensitive manner. However, NFATc.beta neither bound to the kappa3 element (an NFAT-binding site) in the tumor necrosis factor-alpha promoter nor activated the tumor necrosis factor-alpha promoter in cotransfection assays. These data suggest that different members or isoforms of NFAT gene family may regulate inducible expression of different cytokine genes. PMID- 8702850 TI - A functional phosphatidylinositol 3,4,5-trisphosphate/phosphoinositide binding domain in the clathrin adaptor AP-2 alpha subunit. Implications for the endocytic pathway. AB - Clathrin-coated pits are sites of concentration of ligand-bound signaling receptors. Several such receptors are known to recruit, bind, and activate the heterodimeric phosphatidylinositol-3-kinase, resulting in the generation of phosphatidylinositol 3,4, 5-trisphosphate. We report here that dioctanoyl phosphatidylinositol-3,4,5-P3 binds specifically and saturably to soluble AP-2 and with greater affinity to AP-2 within assembled coat structures. Soluble -myo inositol hexakisphosphate shows converse behavior. Binding to bovine brain clathrin-coated vesicles is evident only after detergent extraction. These observations and evidence for recognition of the diacylglyceryl backbone as well as the inositol phosphate headgroup are consistent with AP-2 interaction with membrane phosphoinositides in coated vesicles and with soluble inositol phosphates in cytoplasm. A discrete binding domain is identified near the N terminus of the AP-2 alpha subunit, and an expressed fusion protein containing this sequence exhibits specific, high affinity binding that is virtually identical to the parent protein. This region of the AP-2 alpha sequence also shows the greatest conservation between a Caenorhabditis elegans homolog and mammalian alpha, consistent with a function in recognition of an evolutionarily unchanging low molecular weight ligand. Binding of phosphatidylinositol 3,4, 5 trisphosphate to AP-2 inhibits the protein's clathrin binding and assembly activities. These findings are discussed in the context of the potential roles of phosphoinositides and AP-2 in the internalization and trafficking of cell surface receptors. PMID- 8702851 TI - Activation of a calcium-calmodulin-dependent protein kinase I cascade in PC12 cells. AB - It has been observed that the activity of Ca2+-calmodulin (CaM)-dependent protein kinase I is enhanced up to 50-fold by its phosphorylation in vitro by a distinct CaM kinase I kinase (Lee, J. C., and Edelman, A. M. (1994) J. Biol. Chem. 269, 2158-2164). It has, however, been unclear whether this event represents an acute form of cellular regulation. We demonstrate here the phosphorylation and activation of CaM kinase I in PC12 pheochromocytoma cells in response to elevation of intracellular Ca2+. Treatment of PC12 cells with the Ca2+-ionophore, ionomycin, or with a depolarizing concentration of KCl, led to rapid, biphasic phosphorylation of CaM kinase I and to increases in CaM kinase I activity of 5.1- and 7. 3-fold, respectively. Depolarization-induced activation of CaM kinase I was reduced by approximately 80% by blockade of Ca2+ influx through L-type voltage-dependent Ca2+ channels and completely abolished by removal of extracellular Ca2+. The ability of PC12 cell CaM kinase I to be phosphorylated and activated by purified CaM kinase I kinase in vitro was markedly reduced by prior depolarization of the cells, consistent with intracellular phosphorylation and activation of CaM kinase I by CaM kinase I kinase. These results demonstrate the existence in PC12 cells of a CaM kinase I cascade, the function of which may be to sensitize cells to signal-induced elevations of intracellular Ca2+. PMID- 8702852 TI - Requirement of lysine residues outside of the proposed pentasaccharide binding region for high affinity heparin binding and activation of human antithrombin III. AB - Variant forms of human antithrombin III with glutamine or threonine substitutions at Lys114, Lys125, Lys133, Lys136, and Lys139 were expressed in insect cells to evaluate their roles in heparin binding and activation. Recombinant native ATIII and all of the variants had very similar second order rate constants for thrombin inhibition in the absence of heparin, ranging from 1.13 x 10(5) M-1min-1 to 1.66 x 10(5) M-1min-1. Direct binding studies using 125I-flouresceinamine-heparin yielded a Kd of 6 nM for the recombinant native ATIII and K136T, whereas K114Q and K139Q bound heparin so poorly that a Kd could not be determined. K125Q had a moderately reduced affinity. Heparin binding affinity correlated directly with heparin cofactor activity. Recombinant native ATIII was nearly identical to plasma-purified ATIII, whereas K114Q and K139Q were severely impaired in heparin cofactor activity. K125Q and K136T were only slightly impaired. Based on these data, Lys114 and Lys139, which are outside of the putative pentasaccharide binding site, play pivotal roles in the high affinity binding of heparin to ATIII and the activation of thrombin inhibitory activity. PMID- 8702853 TI - The e subunit gene of murine F1F0-ATP synthase. Genomic sequence, chromosomal mapping, and diet regulation. AB - Genomic sequences encoding murine Lfm1, whose predicted protein sequence is 96% and 98% similar to bovine and rat F1F0-ATP synthase e subunits (respectively), have been amplified from BALB/cByJ DNA, cloned, and sequenced. The 1.1-kilobase gene has 3 introns and 4 exons, and its coding sequence differs by two nucleotides compared to the previously published BALB/cHnn Lfm1 cDNA sequence. A PstI restriction site polymorphism in intron 2 between C57BL/6J and Mus spretus was used to map this gene to Chromosome 5 near D5Mit9. Related sequences were mapped on Chromosomes 8, 11, and 2 unlinked loci on Chromosome 2 using Southern blot analyses with the 1. 1-kilobase gene as probe. Previous studies from this laboratory indicated that the Lfm1/e subunit was regulated by the level of dietary fat and carbohydrate. Northern hybridization analyses demonstrated that e subunit mRNA abundance showed statistically significant differences (p < 0.025) between hearts of BALB/c mice fed 3% and those fed 20% corn oil for 2 weeks and in liver (p < 0. 05) from the same animals. Significant differences were also observed in hepatic and heart mRNA expression at different times after eating in animals subjected to a fast/refeed regimen. The implications of the high degree of sequence similarity to the e subunit for rat and bovine F1F0-ATP synthase and its regulation by diet are discussed. PMID- 8702854 TI - Purification and molecular cloning of an 8R-lipoxygenase from the coral Plexaura homomalla reveal the related primary structures of R- and S-lipoxygenases. AB - Lipoxygenases that form S configuration fatty acid hydroperoxides have been purified or cloned from plant and mammalian sources. Our objectives were to characterize one of the lipoxygenases with R stereospecificity, many of which are described in marine and freshwater invertebrates. Characterization of the primary structure of an R-specific enzyme should help provide a new perspective to consider the enzyme-substrate interactions that are the basis of the specificity of all lipoxygenases. We purified an 8R-lipoxygenase of the prostaglandin containing coral Plexaura homomalla by cation and anion exchange chromatography. This yielded a colorless enzyme preparation, a band of approximately 100 kDa on SDS-polyacrylamide gel electrophoresis, and turnover numbers of 4000 min-1 of 8R lipoxygenase activity in peak chromatographic fractions. The full-length cDNA was cloned by PCR using peptide sequence from the purified protein and by 5'- and 3' rapid amplification of cDNA ends. The cDNA encodes a polypeptide of 715 amino acids, including over 70 amino acids identified by peptide microsequencing. A peptide presequence of 52 amino acids is cleaved to give the mature protein of 76 kDa; the difference from the estimated size by SDS-PAGE implies a post translational modification of the P. homomalla enzyme. All of the iron-binding histidines of S-lipoxygenases are conserved in the 8R-lipoxygenase. However, the C-terminal amino acid is a threonine, as opposed to the isoleucine that provides the carboxylate ligand to the iron in all known S-lipoxygenases. These results establish that the 8R-lipoxygenase is related in primary structure to the S lipoxygenases. A model of the basis of R and S stereospecificity is described. PMID- 8702855 TI - The development and use of a DNA polymerase arrest assay for the evaluation of parameters affecting intrastrand tetraplex formation. AB - We show here that a K+-dependent block to DNA synthesis is a sensitive and specific indicator of intrastrand tetraplex formation that can be used, both to identify sequences with tetraplex-forming potential and to examine parameters that affect tetraplex formation. We show that tetraplex formation is determined by a complex combination of factors including the size and base composition of its constituent loops and stems. In the process of carrying out this study we have found that the number of sequences with the ability to form tetraplexes is larger than previously thought, and that such sequences are ubiquitous in eukaryote genomes. PMID- 8702856 TI - Cloning and characterization of an alternatively processed human type II interleukin-1 receptor mRNA. AB - Two types of interleukin (IL)-1 receptors with three extracellular immunoglobulin like domains, limited homology (28%), and different pharmacological characteristics termed type I and type II have been cloned from mouse and human cell lines. Both receptors exist in transmembrane and soluble forms; the soluble IL-1 receptor is thought to be post-translationally derived from cleavage of the extracellular portion of the membrane receptors. In preliminary cross-linking studies with radiolabeled IL-1, we found that monkey kidney COS1 cells express a soluble receptor with molecular mass of approximately 55-60 kDa, which is different from previously reported soluble IL-1 receptors. This soluble IL-1 receptor protein from COS1 cells was purified to homogeneity by affinity chromatography using recombinant IL-1beta as the ligand and shown to have an affinity for human 125I-IL-1beta (KD approximately 2-3 nM) comparable to the human type II IL-1 receptor (IL-1RII). The purified protein was microsequenced, and the sequence information was used to design primers to clone the COS1 IL-1RII using reverse transcription-coupled polymerase chain reaction; the DNA comparison with monkey COS1 and human IL-1RII indicate that they are 95% identical at the nucleic acid and amino acid levels. In addition, another cDNA, which represents an alternatively processed mRNA of the IL-1RII gene, was also cloned both from monkey COS1 and human Raji cells and was shown to have approximately 95% sequence identity between these species. While the cDNA of the novel alternatively processed gene has a 5' end identical to the IL-1RII, the 200 base pairs at the 3' end are different and the sequence predicts a soluble IL-1 receptor protein of 296 amino acids. Radioligand binding studies of the alternatively processed IL 1RII mRNA demonstrated kinetic and pharmacological characteristics similar to the known type II IL-1 receptor. COS7 cells (which lack IL-1 receptor) transfected with the transmembrane form of the human IL-1RII cDNA showed 125I-IL-1beta binding in both the membrane fractions and supernatant. In contrast, COS7 cells transfected with the alternatively processed human IL-1RII cDNA showed high affinity 125I-IL-1beta binding (Ki approximately 1.2 nM) predominantly in the supernatant; a very small amount of detectable membrane IL-1 binding activity was also observed presumably due to association of the soluble IL-1 receptor and membrane-integrated proteins. In cross-linking and ligand blot studies, the alternatively processed human IL-1RII cDNA-transfected COS7 cells expressed a soluble IL-1 receptor with molecular masses ranging from 60 to 160 kDa, further indicating the association between this soluble IL-1 receptor and other soluble proteins. In summary, we report the purification and characterization of a soluble IL-1 receptor expressed by COS1 cells and the cloning of an alternatively processed type II IL-1 receptor mRNA from both human and COS1 cells. The alternative splicing of a primary transcript leading to a secreted protein provides a potentially important mechanism by which soluble IL-1RII can be produced. PMID- 8702857 TI - Ubiquitination of protein kinase C-alpha and degradation by the proteasome. AB - Bryostatins and phorbol esters acutely activate and subsequently down-regulate protein kinase C (PKC) by inducing its proteolysis via an unknown pathway. Here we show that treatment of renal epithelial cells with bryostatin 1 (Bryo) produced novel PKC-alpha species, which were larger than the native protein (80 kDa). The >80 kDa PKC-alpha species contained Ubi as indicated by immunostaining and accumulated in the presence of lactacystin, a selective inhibitor of proteolysis by the proteasome. In vitro experiments with 125I-ubiquitin and membranes from Bryo-treated cells showed that PKC-alpha became ubiquitinated by a reaction that depended on ATP and a cytosolic fraction. Lactacystin or a peptidyl aldehyde, Bz-Gly-Leu-Ala-leucinal, which inhibits certain proteinase activities of the proteasome, inhibited Bryo-evoked disappearance of PKC-alpha protein from the cells. Lacta preserved Bryo-induced 32P-labeled PKC-alpha indicating that the proteasome inhibitor spared activated enzyme from down-regulation in vivo. These findings show that Bryo induces the degradation of PKC-alpha by the ubiquitin proteasome complex. PMID- 8702858 TI - Molecular ordering of apoptotic mammalian CED-3/ICE-like proteases. AB - Apoptosis is executed by cysteine proteases belonging to the CED-3/ICE family, which, unlike other mammalian cysteine proteases, cleave their substrates following aspartate residues. Proteases belonging to this family exist in the cytosol as zymogens that require accurate processing at internal aspartate residues to generate the two-chain active enzymes. As such, CED-3/ICE family members are capable of activating each other in a manner analogous to the protease zymogens of the coagulation or complement cascades. At present, it is unknown whether such mutual processing exists in vivo, and if so whether it is sequential, implying an order to the death pathway. Using a cell-free apoptosis system, recombinant ICE proteases and both biochemical and morphological criteria, we demonstrate an ordering of the mammalian ICEs that are most related to the Caenorhabditis elegans death protease CED-3. PMID- 8702859 TI - Epidermal growth factor induces coupling of protein-tyrosine phosphatase 1D to GRB2 via the COOH-terminal SH3 domain of GRB2. AB - The Src homology 2 (SH2) and SH3 domain-containing adaptor protein GRB2 and the SH2 domain-containing protein-tyrosine phosphatase 1D (PTP1D, also called SHPTP2, PTP2C, SHPTP3, Syp, or SHP-2) function as positive mediators of growth factor induced mitogenesis. Epidermal growth factor (EGF) is a potent mitogen for MCF 10A human mammary epithelial cells and EGF receptor-expressing mouse NR6 fibroblasts. Western blot analysis of anti-PTP1D immune complexes derived from EGF-treated cells demonstrated a ligand-dependent coupling between the phosphatase and GRB2 in vivo. Probing of lysates from these cells with glutathione S-transferase (GST) fusion proteins corresponding to the individual domains of GRB2 revealed that this interaction was mediated exclusively by the COOH-terminal SH3 domain of GRB2. Importantly, a GST fusion protein containing the PTP1D SH2 domains was not capable of generating the EGF-induced linkage to GRB2. Additional experiments indicated that neither the binding of the nucleotide exchange factor Sos to GRB2 nor tyrosine phosphorylation of PTP1D was required for EGF-stimulated coupling of PTP1D to GRB2. This is the first demonstration of a growth factor- or cytokine-induced coupling of a protein through an SH3 domain and suggests that GRB2 functions to target PTP1D, in addition to Sos, to the plasma membrane in response to EGF. PMID- 8702860 TI - Enhanced thrombin sensitivity of a factor VIII-heparin cofactor II hybrid. AB - Generation of thrombin at a site of vascular injury is a key event in the arrest of bleeding. In addition to the conversion of fibrinogen into the insoluble fibrin, thrombin can initiate a number of positive and negative feedback mechanisms that either sustain or down-regulate clot formation. We have modulated the thrombin sensitivity of human blood coagulation factor VIII, an essential cofactor in the intrinsic pathway of blood coagulation. We have substituted an acidic region of factor VIII corresponding to amino acid sequence Asp712-Ala736 by amino acid sequence Ile51-Leu80 of the thrombin inhibitor heparin cofactor II. Functional analysis of the resulting factor VIII-heparin cofactor II hybrid, termed des-(868-1562)-factor VIII-HCII, revealed an increase in procoagulant activity as measured in a one-stage clotting assay. Incubation of purified des (868-1562)-factor VIII-HCII with different amounts of thrombin showed that this protein was more readily activated by thrombin when compared with des-(868-1562) factor VIII, a control protein lacking amino acid sequence Ile51-Leu80 of heparin cofactor II. This was manifested by an increase in the second order rate constant of activation by thrombin for des-(868-1562)-factor VIII-HCII (12.0 +/- 0.48 x 10(6) M-1 s-1) compared with des-(868-1562)-factor VIII (1.77 +/- 0.21 x 10(6) M 1 s-1). Our data suggest that amino acid sequence Ile51-Leu80 of heparin cofactor II endows factor VIII with increased sensitivity towards thrombin which results in accelerated clot formation. PMID- 8702861 TI - The calcium-dependent association and functional coupling of calmodulin with myocardial phospholipase A2. Implications for cardiac cycle-dependent alterations in phospholipolysis. AB - Herein we demonstrate the calcium-dependent regulation of myocardial phospholipase A2 activity, which is mediated by a cytosolic protein constituent that can be chromatographically resolved from, and subsequently reconstituted with, purified myocardial phospholipase A2. Purification of this protein by sequential column chromatographies revealed an 18-kDa doublet, which was identified as calmodulin by Western blotting, calcium-dependent precipitation with W-7 agarose beads, and reconstitution of calcium-mediated phospholipase A2 inhibition with authentic homogeneous calmodulin. Calcium-induced calmodulin mediated inhibition of myocardial phospholipase A2 was titrated by physiologic increments of calcium ion (Kd approximately 200 nM). Moreover, ternary complex affinity chromatography with calmodulin-Sepharose demonstrated that inhibition of myocardial phospholipase A2 activity by calmodulin resulted from the direct interaction of calmodulin with the myocardial phospholipase A2 catalytic complex. Exposure of cultured A-10 muscle cells to three structurally disparate calmodulin antagonists (W-7, trifluoperazine, and calmidazolium) resulted in the robust release of arachidonic acid, which was entirely ablated by pretreatment of cells with (E)-6-(bromomethylene)-3-(1-naphthalenyl)-2-H-tetrahydropyran-2-one. Collectively, this study identifies a novel mechanism whereby latent phospholipase A2 activity can be released from tonic inhibition by alterations in the interactions between the phospholipase A2 catalytic complex, calcium ion, and the intracellular calcium transducer, calmodulin. PMID- 8702862 TI - Homeodomain interaction with the beta subunit of the general transcription factor TFIIE. AB - Homeodomain-containing proteins play a crucial role as transcriptional regulators in the process of cell differentiation. The homeodomain performs a dual function in this regard, acting as a DNA binding domain and participating in protein protein interactions that enhance DNA binding specificity or regulatory activity. Here we describe a homeodomain class-specific interaction with the beta subunit of the general transcription factor TFIIE. We show that the Antennapedia and Abdominal-B homeodomains bind to TFIIEbeta, but the even-skipped homeodomain does not. Using a two-hybrid assay performed in cultured cells, we demonstrate that the homeodomain-TFIIEbeta interaction occurs in vivo. The Abdominal-B homeodomain is shown to activate transcription in vitro, and this activation can be blocked with anti-TFIIEbeta antibody without affecting basal transcription levels. Together with published data demonstrating an interaction between proteins containing even-skipped class homeodomains and the TATA-binding protein (Um, M., Li, C., and Manley, J. L. (1995) Mol. Cell. Biol. 15, 5007-5016; Zhang, H., Catron, K. M., and Abate-Shen, C. (1996) Proc. Natl. Acad. Sci. U. S. A. 93, 1764 1769), these results suggest various homeodomain containing proteins interact with different general transcription factors, an observation that may have important implications for transcriptional regulation. PMID- 8702863 TI - Chemotactic peptide-induced activation of MEK-2, the predominant isoform in human neutrophils. Inhibition by wortmannin. AB - Exposure of neutrophils to a variety of agonists including chemoattractant peptides and cytokines induces degranulation and activation of the oxidative burst which are required for bacterial killing. The signaling pathways regulating these important functions are incompletely characterized. Mitogen-activated protein (MAP) kinases, which include the extracellular signal-regulated kinases (ERKs), are activated rapidly in neutrophils, suggesting that they may regulate cell activation. We found that neutrophils express two isoforms of MAP/ERK kinase (MEK), mixed-function kinases that are responsible for phosphorylation and activation of ERK. Like MEK-1, MEK-2 was found to reside in the cytosol both before and after stimulation. Studies were undertaken to define the relative abundance and functional contribution of MEK-1 and MEK-2 in neutrophils and to characterize the signaling pathways leading to their activation. Although the abundance of the two isoforms was similar, the activity of MEK-2 was at least 3 fold greater than that of MEK-1. A rise in cytosolic [Ca2+] was insufficient for MEK stimulation, and blunting the [Ca2+] change with intracellular chelators failed to prevent receptor-mediated activation of either isoform, implying that cytosolic Ca2+ transients are not necessary. In contrast, both MEK-1 and MEK-2 were activated by exposure of cells to protein kinase C (PKC) agonists. Conversely, PKC antagonists inhibited the chemotactic stimulation of both isoforms, suggesting that PKC was required for their activation. Despite these similarities, clear differences were also found in the pathways leading to activation of the MEK isoforms. In particular, MEK-2 was considerably more sensitive than MEK-1 to the phosphatidylinositol 3-kinase inhibitor wortmannin. Phosphorylation and activation of ERK-1 and ERK-2 were also reduced by this inhibitor. In summary, MEK-2 is stimulated in formyl-methionyl-leucyl phenylalanine-treated neutrophils, where it appears to be functionally the predominant isoform. The time course and inhibitor sensitivity of MEK-2 activation parallel those of several components of the microbicidal response, suggesting a signaling role of the MEK-ERK pathway. PMID- 8702864 TI - Characterization of three potato lipoxygenases with distinct enzymatic activities and different organ-specific and wound-regulated expression patterns. AB - Lipoxygenases are ubiquitous enzymes in eukaryotes. In plants, lipoxygenases are involved in the synthesis of the hormone jasmonic acid that regulates plant responses to wounding and, in addition, is an inducer of tuberization in potato. We have isolated potato lipoxygenase cDNA clones. From their deduced amino acid sequences, three distinct classes are defined (Lox1, Lox2, and Lox3). They are encoded in gene families that display organ-specific expression, lox1 being expressed mostly in tubers and roots, lox2 in leaves, and lox3 in leaves and roots. Consistent with their organ-specific expression pattern, Lox1 expressed in bacteria preferentially uses as substrate linoleic acid, abundant in membrane lipids of tubers, whereas linolenic acid, prevalent in leaves, is the preferred substrate for the other two classes of lipoxygenase. Analyses on reaction products of the enzymes expressed in bacteria reveal that Lox1 primarily produces 9- hydroperoxides. In contrast, the jasmonic acid precursor, 13 hydroperoxylinolenic acid, is the major product of the action of Lox2 and Lox3 on linolenic acid. Upon wounding, the levels of Lox2 and Lox3 transcripts rise markedly in leaves. While Lox3 mRNA accumulation peaks as early as 30 min after wounding, Lox2 shows a steady increase over a 24-h time course, suggesting different roles for these lipoxygenase isoforms in the synthesis of the plant hormone jasmonic acid. PMID- 8702865 TI - Inhibitor probes of the quinone binding sites of mammalian complex II and Escherichia coli fumarate reductase. AB - The structural and catalytic properties of beef heart succinate dehydrogenase (succinate-ubiquinone oxidoreductase, complex II) and Escherichia coli fumarate reductase are remarkably similar. One exception is that whereas electron exchange between the mammalian enzyme and its quinone pool is inhibited by thenoyltrifluoroacetone and carboxanilides, the enzyme from E. coli is not sensitive to these inhibitors. The lack of good inhibitors has seriously hampered the elucidation of the mechanism of quinone oxidation/reduction in the E. coli enzyme. We have previously reported (Tan, A. K., Ramsay, R. R., Singer, T. P., and Miyoshi, H. (1993) J. Biol. Chem. 268, 19328-19333) that 2-alkyl-4,6 dinitrophenols inhibit mammalian complexes I, II, and III, but with different potencies and kinetic characteristics. Based on these studies we have selected a series of 2-alkyl-4,6-dinitrophenols which proved to be very effective noncompetitive inhibitors of mammalian complex II, particularly when acting in the direction of quinone reduction, the physiological event. These compounds turned out to be even more potent inhibitors of E. coli fumarate reductase, particularly when acting in the direction of quinol oxidation, again, the physiological event. Kinetic analysis revealed that with both enzymes 2 inhibitor binding sites seem to be involved in the oxidation of succinate by quinone, but one seems to be functioning when fumarate is reduced by external quinol. Since the E. coli enzyme can be modified by site-directed mutagenesis, these studies were extended to four mutants of fumarate reductase, impaired by single amino acid substitutions at either of the putative quinone binding sites (QA or QB) of the enzyme. The results were analyzed in terms of the model of these dual sites of quinone binding in fumarate reductase, as well as the nature of the substituent in the 2-position of the dinitrophenol inhibitors. PMID- 8702866 TI - The dinuclear copper site structure of Agaricus bisporus tyrosinase in solution probed by X-ray absorption spectroscopy. AB - We have measured the x-ray absorption near edge structure (XANES) spectra of the enzyme tyrosinase from the mushroom Agaricus bisporus in solution in the oxy and deoxy forms. The spectra, obtained under the same conditions as the analogous forms of mollusc hemocyanin (Hc), show that the oxidation state of copper changes from Cu(II) (oxy form) to Cu(I) (deoxy form), and the copper active site(s) of A. bisporus tyrosinase in solution undergoes the same main conformational changes as Hc. We have applied the multiple scattering theory to simulate the XANES spectra of various alternative geometries of the copper site, accounting for the residual differences between Hc and tyrosinase. While oxy-Hc is reasonably fitted only by the pseudo-square-pyramidal geometry reported by its crystallographic data, oxytyrosinase can be fitted, starting from the Hc coordinates, either by distortions toward a pseudo-tetrahedral geometry, with inequivalent copper sites, or by an apically distorted square-pyramidal geometry (with an elongation of the apical distance of no more than 0.2 A). PMID- 8702867 TI - The RNA subunit of ribonuclease P from the zebrafish, Danio rerio. AB - A simple strategy has been devised to identify the gene encoding the RNA subunit of RNase P from the zebrafish, Danio rerio. The sequence obtained by amplification of genomic DNA with primers based on sequences common to two other vertebrates was confirmed by reverse transcription and amplification of RNA from a partially purified preparation of the holoenzyme. The 5' and 3' ends were determined by cyclizing the RNA, followed by reverse transcription and sequencing across the ligated RNA junction. The zebrafish sequence is 63% identical to that of Xenopus laevis nuclear RNase P RNA and 69% identical to the human RNase P RNA. A consensus secondary structure was constructed based on these nucleotide identities and on the many compensatory base changes in several regions among these three RNAs. The strategy used to obtain the zebrafish sequence should be useful in deriving analogous gene sequences from diverse classes of eukaryotes. PMID- 8702868 TI - The mechanism of the auxotrophy for sulfur-containing amino acids imposed upon Escherichia coli by superoxide. AB - Defects in both of the genes coding for the cytosolic superoxide dismutases (SODs) of Escherichia coli impose an oxygen-dependent nutritional requirement for cysteine. This is now seen to be a bradytrophy, rather than an absolute auxotrophy, since lack of Cys merely imposed a growth lag and escape from this growth lag did not involve genetic reversion. This Cys bradytrophy was not seen in the SOD-competent parental strain, and it was relieved by a cell-permeant mimic of SOD activity; hence, it was due to O2-.. It was also relieved by an osmolyte, such as sucrose; hence, it appears due to leakage from the cell of some component needed for Cys biosynthesis. Medium conditioned by the aerobic growth of the SOD-defective strain relieved the growth lag. Bioassays with Cys mutants suggested that the conditioned medium contained SO3-3 or its equivalent, and sulfite per se was able to eliminate the growth lag. However, some component of the conditioned medium reacted with added sulfite and interfered with attempts to assay for it colorimetrically. These results suggest that the cell envelope of the SOD-defective strain was weakened, directly or indirectly, by O2 and then leaked sulfite. This prevents cysteine biosynthesis until sulfite accumulates in the medium. PMID- 8702869 TI - The role of positive charges and structural segments in the presequence of rat liver aldehyde dehydrogenase in import into mitochondria. AB - Most mitochondrial proteins are nucleus-encoded and translated in the cytosol. They have an N-terminal presequence that allows recognition by the mitochondrial import apparatus and subsequent import into mitochondria. These presequences are rich in positive charges, mainly arginines. The role of these positive charges in the 19-amino acid presequence of rat liver aldehyde dehydrogenase was investigated by systematically replacing them with the polar but uncharged residue, glutamine. The single substitution of any of the four Arg residues in the helical segments did not affect import. Substitution of both Arg residues in the N-terminal segment (R3Q/R10Q) caused a dramatic decrease in import competence. This could be restored by using the mutant lacking the three-amino acid (RGP) linker that separates the two helical domains, determined by two dimensional NMR (Thornton, K., Wang, Y., Weiner, H., and Gorenstein, D. G. (1993) J. Biol. Chem. 268, 19906-19914). CD and NMR spectra of the peptide corresponding to the linker-deleted presequence showed that it was substantially more prone to helix formation than the native peptide over its entire length. A similar analysis of the peptide corresponding to the R3Q/R10Q presequence revealed that this peptide was only somewhat more helical than the native peptide and that the greater helicity did not include the residues near the N terminus. It is concluded that positively charged residues in the presequence play a vital role in the import of precursor aldehyde dehydrogenase. One of the positive charges in the N-terminal helical segment of the presequence is necessary for import competence. However, if both positive charges are removed, import competence can be retained as long as the presequence is capable of forming a relatively more stable alpha-helix near its N terminus. PMID- 8702870 TI - The Ste20-like protein kinase, Mst1, dimerizes and contains an inhibitory domain. AB - The human serine/threonine protein kinases, Mst1 and Mst2, share considerable homology to Ste20 and p21-activated kinase (Pak) throughout their catalytic domains. However, outside the catalytic domains there are no significant homologies to previously described Ste20-like kinases or other proteins. To understand the role of the nonhomologous regions, we performed a structure/function analysis of Mst1. A series of COOH-terminal and internal deletions indicates that there is an element within a central 63-amino acid region of the molecule that inhibits kinase activity. Removal of this domain increases kinase activity approximately 9-fold. Coimmunoprecipitation assays, the yeast two-hybrid procedure, and in vitro cross-linking analysis indicate that Mst1 homodimerizes and that the extreme COOH-terminal 57 amino acids are required for self-association. Size exclusion chromatography indicates that Mst1 is associated with a high molecular weight complex in cells, suggesting that other proteins may also oligomerize with this kinase. While loss of dimerization alone does not affect kinase activity, a molecule lacking both the dimerization and inhibitory domains is not as active as one which lacks only the inhibitory domain. Comparison of Mst1 and Mst2 indicates that both functional domains lie in regions conserved between the two molecules. PMID- 8702871 TI - Active site structural features for chemically modified forms of rhodanese. AB - In the course of the reaction catalyzed by rhodanese, the enzyme cycles between two catalytic intermediates, the sulfur-free and the sulfur-substituted (persulfide-containing) forms. The crystal structure of sulfur-free rhodanese, which was prepared in solution and then crystallized, is highly similar to that of sulfur-substituted enzyme. The inactivation of sulfur-free rhodanese with a small molar excess of hydrogen peroxide relies essentially on a modification limited to the active site, consisting of the oxidation of the essential sulfhydryl to sulfenyl group (-S-OH). Upon reaction of the sulfur-free enzyme with monoiodoacetate in the crystal, the Cys-247 side chain with the bound carboxymethyl group is forced into a conformation that allows favorable interactions of the carboxylate with the four peptide NH groups that participate in hydrogen bonding interactions with the transferable sulfur atom of the persulfide group in the sulfur-substituted rhodanese. It is concluded that active site-specific chemical modifications of sulfur-free rhodanese do not lead to significant changes of the protein structure, consistent with a high degree of similarity of the structures of the sulfur-free and sulfur-substituted forms of the enzyme both in solution and in the crystal. PMID- 8702872 TI - RRN11 encodes the third subunit of the complex containing Rrn6p and Rrn7p that is essential for the initiation of rDNA transcription by yeast RNA polymerase I. AB - A new gene, RRN11, has been defined by certain rrn mutants of Saccharomyces cerevisiae which are defective specifically in the transcription of 35 S rRNA gene by RNA polymerase I (pol I). We have cloned the gene and found that it encodes a protein of 507 amino acids. We have used a strain with the chromosomal RRN11 deleted and carrying HA1 epitope-tagged RRN11 on a plasmid to isolate a protein complex containing the protein encoded by RRN11. This protein complex complemented rrn6 mutant extracts, which were previously shown to be deficient in the essential pol I transcription factor called Rrn6/7 complex or core factor (CF). The CF complex was previously shown to consist of three proteins, the 102- and 60-kDa subunits encoded by RRN6 and RRN7, respectively, and the 66-kDa subunit. The results of the above complementation experiments combined with mobility of Rrn11p in SDS-polyacrylamide gel electrophoresis analysis relative to Rrn6p and Rrn7p led to the conclusion that RRN11 encodes the 66-kDa subunit of CF. Glutathione S-transferase-Rrn11p fusion protein was found to bind strongly to 35S-labeled Rrn6p and Rrn7p but only weakly to 35S-labeled TATA-binding protein. Similarly, glutathione S-transferase-Rrn7p fusion protein bound strongly to 35S labeled Rrn6p and Rrn11p but only weakly to 35S-labeled TATA-binding protein. These results are consistent with the fact that one can purify CF consisting of Rrn6p, Rrn7p, and Rrn11p from yeast cell extracts, but the purified complex does not contain TATA-binding protein. RRN11 was shown to be an essential gene, and [3H]uridine pulse experiments demonstrated directly that RRN11 is essential for rDNA transcription by pol I in vivo. Thus all three subunits of CF are essential for rDNA transcription. Because of the resemblance of CF to mammalian essential pol I transcription factor SL1, the amino acid sequences of Rrn11p and the other two subunits of CF were compared with those of the three TATA-binding protein associated factors (TAFs) in the human SL1, TAFI48, TAFI63, and TAFI110. No significant similarity was detected between two sets of the proteins. Similarity as well as differences between CF and SL1 are discussed. PMID- 8702873 TI - Multiexon deletions in the type I collagen COL1A2 gene in osteogenesis imperfecta type IB. Molecules containing the shortened alpha2(I) chains show differential incorporation into the bone and skin extracellular matrix. AB - Osteogenesis imperfecta (OI) type IB is a rare subset of the mildest form of OI, clinically characterized by moderate bone fragility, blue sclera, and dentinogenesis imperfecta. Cultured skin fibroblasts from two unrelated individuals (OI-197 and OI-165) with the typical features of OI type IB produced shortened alpha2(I) chains. Reverse transcription-polymerase chain reaction of the alpha2(I)-cDNA revealed deletions in the triple helical domain of 5 exons (exons 7-11) in OI-197, and 8 exons (exons 10-17) in OI-165. This exon skipping was caused by genomic deletions in one allele of COL1A2 with the breakpoints located in introns 6 and 11 in OI-197, and introns 9 and 17 in OI-165. The secretion and deposition of the mutant collagen into the matrix was measured in vitro in cultures of skin fibroblasts and bone osteoblasts, grown in the presence of ascorbic acid to induce collagen matrix formation and maturation, as well as in collagen extracts from skin and bone. The secretion of mutant collagen was impaired and long term cultures of fibroblasts showed that the mutant collagen was not incorporated into the mature collagenous matrix produced in vitro by skin fibroblasts from both patients. Likewise, the shortened alpha2(I) chain was not demonstrable in skin extracts. In contrast, bone extracts from OI-197 showed the presence of the mutant collagen. This incorporation of the abnormal collagen into the mature matrix was also demonstrated in long term cultures of the patient's osteoblasts. The deposition of the mutant collagen by bone osteoblasts but not by skin fibroblasts demonstrates a tissue specificity in the incorporation of mutant collagen into the matrix which may explain the primary involvement of bone and not skin in these patients. PMID- 8702874 TI - Limited proteolysis of rat phosphatidylinositol transfer protein by trypsin cleaves the C terminus, enhances binding to lipid vesicles, and reduces phospholipid transfer activity. AB - Rat phosphatidylinositol transfer protein (PITP) is a 32-kDa protein of 271 amino acids that transfers phosphatidylinositol and phosphatidylcholine between membranes. The alpha isoform of rat PITP was expressed in Escherichia coli and purified in high yields. The purified protein contained 1 mol of phosphatidylglycerol and had a transfer activity for phosphatidylinositol and phosphatidylcholine equal to or greater than that of PITP purified from mammalian brain. Limited protease digestion was used to further define structure, activity, and function relationships in PITP. PITP alone is relatively resistant to digestion by chymotrypsin, trypsin, and Staphylococcus V8 protease but is readily cleaved by subtilisin. Phospholipid vesicles containing phosphatidic acid enhance susceptibility to digestion by all four proteases. In the presence of vesicles, PITP, which migrates as a 36-kDa protein in SDS-polyacrylamide gel electrophoresis, is cleaved rapidly by trypsin to a form that appears to be 2-3 kDa smaller than the native form. The tryptic fragment retains partial phospholipid transfer activity and shows an enhanced affinity for phospholipid vesicles containing phosphatidic acid. Analysis of the tryptic digestion products by immunoblotting, N-terminal sequencing, and electrospray mass spectrometry showed that trypsin cleaves the C terminus of PITP at Arg253 and Arg259. Thus, removal of the C terminus enhances the affinity of PITP for vesicles and results in a dimunition of transfer activity. Overall, the data show that PITP undergoes conformation changes and that the C terminus becomes more accessible to trypsin when bound to vesicles. Hence, the C terminus is not an essential component of the membrane binding site and may be located distal to it. PMID- 8702875 TI - Galpha12 and Galpha13 regulate extracellular signal-regulated kinase and c-Jun kinase pathways by different mechanisms in COS-7 cells. AB - Many growth factors and agonists for G protein-coupled receptors activate mitogen activated protein (MAP) kinase pathways, including the extracellular signal regulated kinase (ERK) pathway and the c-Jun kinase (JNK) pathway. Transient transfection of dominant negative and constitutively active pathway components in COS-7 cells shows that two G protein subunits, Galpha12 and Galpha13, inhibit the ERK pathway and stimulate the JNK pathway. Constitutively active (GTPase deficient) Galpha12 and Galpha13 both inhibit ERK pathway activation by epidermal growth factor. A Galpha13/alphaz chimera, which responds to stimulation by Gi coupled receptors, mediates inhibition of ERK via such a receptor, the dopamine-2 receptor. In addition, expression of a dominant negative mutant of the GTPase, Cdc42, blocks activation of the JNK pathway by Galpha12 and Galpha13 but does not alter inhibition of ERK activation by the same Galpha proteins; conversely, mutationally activated Cdc42 stimulates the JNK pathway but has no effect on the ERK pathway. Our results show that different mechanisms mediate two effects of Galpha12 and Galpha13: the ERK pathway inhibition is mediated at the level of MAP kinase kinase in a Ras- and Raf-independent fashion, whereas the JNK pathway stimulation is mediated by Cdc42. PMID- 8702876 TI - Mapping of Pax-2 transcription activation domains. AB - Pax genes encode transcription factors known to play crucial roles during the development of specific embryonic tissues. In humans and mice, several abnormalities have been linked to deficiencies in Pax gene dosage, indicating that normal development is particularly sensitive to the level of Pax gene expression. Despite these facts, relatively little is known about how these proteins act as transcriptional regulators. In this study we define the transactivation domains of murine Pax-2, an essential factor in kidney organogenesis. Within the COOH terminus of Pax-2, amino acids 279-373 are essential for transactivation. However, this region alone is insufficient for full transactivation when fused to the paired domain alone or to a heterologous DNA binding domain. Mutation or deletion of the conserved octapeptide sequence results in increased transactivation by Pax proteins. The octapeptide-mediated repression is also seen within a heterologous context using the GAL4 DNA binding domain. Thus transactivation by Pax-2 relies upon several regions within the COOH terminus and is down-modulated by the octapeptide element. PMID- 8702877 TI - Characterization of the human forkhead gene FREAC-4. Evidence for regulation by Wilms' tumor suppressor gene (WT-1) and p53. AB - We describe the cloning and sequence analysis of a nearly full-length cDNA as well as a corresponding 5.2-kilobase pair genomic fragment encoding FREAC-4, a member of the forkhead family of transcription factors. The cDNA is collinear with respect to the coding region of the intronless genomic clone. The conceptual translation product predicts a protein of 465 amino acids with a hyperacidic amino-terminal end, a DNA binding forkhead domain and a carboxyl-terminal part that is rich in homopolymeric runs of prolines and alanines. The transcription start is identified using an RNase protection assay. A 2.7-kilobase pair genomic DNA fragment, located immediately upstream of the translation start, was fused to a luciferase reporter gene. Significant levels of luciferase activity were detected when this construct was transfected into two kidney-derived cell lines, 293 and COS-7 cells, whereas only background reporter gene expression was observed in a cell line of nonkidney origin. Cotransfections with plasmids expressing WT-1, WTAR (a mutated form of WT-1), p53, and a mutated form of p53 revealed a complex pattern of regulation with a 3-fold induction with WT-1, a 7 fold induction with mutated p53, and a 4-fold repression with wild-type p53. A 5' promoter deletion series delimits a DNA fragment necessary for WT-1 inducibility in cotransfection experiments. This fragment is shown to contain at least one cis element that is capable of interacting with recombinant WT-1. PMID- 8702878 TI - Neurotrophins stimulate the release of dopamine from rat mesencephalic neurons via Trk and p75Lntr receptors. AB - We analyzed the short term effect of neurotrophins on mesencephalic neuronal cultures of embryonic (E14) rats with respect to which receptors mediate the actions. Brain-derived neurotrophic factor (BDNF) or neurotrophin-3 enhanced within minutes in a dose-dependent manner (2, 20, 100 ng/ml for 5 min) depolarization-induced (KCl, 30 mM 5 min) and basal dopamine release, but nerve growth factor (NGF) was only effective at high doses (100 ng/ml). The effect of BDNF, but not of NGF, was blocked by K252a or K252b. BDNF, but not NGF, phosphorylated trkB receptors. The NGF-induced, but not the BDNF-induced effect upon the release of dopamine was blocked by anti-p75 antibody MC192. C2-ceramide, an analogue of ceramide, the second messenger of the sphingomyelin pathway, and sphingomyelinase itself induced a release of dopamine comparable with the effect of NGF. NGF, but not BDNF, increased ceramide production. In addition, simultaneous treatment with BDNF and NGF led to a partial prevention of the NGF stimulated, p75(Lntr)-mediated effect. We conclude that BDNF stimulates the release of dopamine by activation of the trkB receptor, whereas NGF affects the release via the p75(Lntr) receptor inducing the sphingomyelin pathway. PMID- 8702879 TI - Site-specific phosphorylation of synapsin I by mitogen-activated protein kinase and Cdk5 and its effects on physiological functions. AB - Posttranslational modifications of synapsin I, a major phosphoprotein in synaptic terminals, were studied by mass spectrometry. In addition to a well known phosphorylation site by calmodulin-dependent protein kinase II (CaM kinase II), a hitherto unrecognized site (Ser553) was found phosphorylated in vivo. The phosphorylation site is immediately followed by a proline, suggesting that the protein is an in vivo substrate of so-called proline-directed protein kinase(s). To identify the kinase involved, three proline-directed protein kinases expressed highly in the brain, i.e. mitogen-activated protein (MAP) kinase, Cdk5-p23, and glycogen synthase kinase 3beta, were tested for the in vitro phosphorylation of synapsin I. Only MAP kinase and Cdk5-p23 phosphorylated synapsin I stoichiometrically. The phosphorylation sites were determined to be Ser551 and Ser553 with Cdk5-p23, and Ser62, Ser67, and Ser551 with MAP kinase. Upon phosphorylation with MAP kinase, synapsin I showed reduced F-actin bundling activity, while no significant effect on the interaction was observed with the protein phosphorylated with Cdk5-p23. These results raise the possibility that the so-called proline-directed protein kinases together with CaM kinase II and cAMP-dependent protein kinase play an important role in the regulation of synapsin I function. PMID- 8702880 TI - Exons lost and found. Unusual evolution of a seminal vesicle transglutaminase substrate. AB - The GP1G gene codes for three of the four abundant androgen-regulated secretory proteins produced by the guinea pig seminal vesicle. Sequencing of the entire 6.3 kilobase gene and comparison with other mammalian seminal vesicle secretory protein genes reveals a common three-exon, two-intron organization. However, significant sequence similarity between this group of genes is largely limited to their 5'-flanking regions and first exons, which code almost exclusively for signal peptides in each case. The first intron of GP1G does contain a region with high similarity to the coding exon of a human seminal vesicle secretory protein gene, semenogelin II. The 3' half of the GP1G gene appears to share a common ancestry with the human SKALP/elafin gene. Sequences related to the elafin promoter, coding, untranslated regions, and introns are clearly identifiable within the GP1G sequence. The elafin gene codes for a serine protease inhibitor and is expressed in a variety of different human tissues. To determine if the GP1G gene was also active outside of the seminal vesicle, RNA from a variety of guinea pig tissues was hybridized to a GP1G cDNA probe. At least three novel RNA bands hybridizing to the GP1G probe were detected in testis RNA samples, and GP1G related mRNAs were also found in other tissues. These data suggest that these seminal vesicle secretory proteins may have functional roles outside the reproductive system. PMID- 8702881 TI - Participation of nucleoside-diphosphate kinase in muscarinic K+ channel activation does not involve GTP formation. AB - Agonist-bound muscarinic receptors open atrial K+ channels through a GTP dependent pathway mediated by the G protein Gk. However, nucleotides other than GTP are also able to support channel activity, even in the absence of agonists. This process was proposed to be mediated by nucleoside-diphosphate (NDP) kinase, which would transfer phosphate from nucleotide triphosphates to the GDP bound to Gk, producing Gk-GTP without the need for receptor-induced GDP-GTP exchange. We examined the effect of antibodies to NDP kinase on the ATP-supported activity of atrial muscarinic K+ channels and the corresponding GIRK1/CIR channels expressed in HEK 293 cells. Inhibitory antibodies reduced ATP-induced channel openings, but this effect displayed an absolute requirement for agonist and was also seen with antibodies that do not inhibit the enzyme. Both types of antibodies also reduced agonist-dependent channel activity in the presence of GTP, ruling out a role for NDP kinase in GDP rephosphorylation. Channel activity was not affected by the antibodies in preparations where ATP-induced muscarinic channels are not under tight receptor control, namely pertussis toxin-treated atrial patches and membranes from cells expressing KACh channel subunits. Thus, participation of NDP kinase in this pathway requires activated receptors and has a function distinct from phosphate transfer between nucleotides. PMID- 8702882 TI - Activation and inhibition of phosphorylase kinase by monospecific antibodies raised against peptides from the regulatory domain of the gamma-subunit. AB - The C terminus of the catalytic gamma-subunit of phosphorylase kinase comprises a regulatory domain that contains regions important for subunit interactions and autoinhibitory functions. Monospecific antibodies raised against four synthetic peptides from this region, PhK1 (362-386), PhK5 (342-366), PhK9 (322-346), and PhK13 (302-326), were found to have significant effects on the catalytic activities of phosphorylase kinase holoenzyme and the gamma delta complex. Antibodies raised against the very C terminus of the gamma-subunit, anti-PhK1 and anti-PhK5, markedly activated both holoenzyme and the gamma delta complex, in the presence and absence of Ca2+. In the presence of Ca2+ at pH 8.2, anti-PhK1 activated the holoenzyme more than 11-fold and activated the gamma delta complex 2.5-fold. Activation of the holoenzyme and the gamma delta complex by anti-PhK5 was 50-70% of that observed with anti-PhK1. Prior phosphorylation of the holoenzyme by the cAMP-dependent protein kinase blocked activation by both anti PhK1 and anti-PhK5. Antibodies raised against the peptides from the N terminus of the regulatory domain, anti-PhK9 and anti-PhK13, were inhibitory, with their greatest effects on the gamma delta complex. These data demonstrate that the binding of antibodies to specific regions within the regulatory domain of the gamma-subunit can augment or inhibit structural changes and subunit interactions important in regulating phosphorylase kinase activity. PMID- 8702883 TI - Interleukin-1beta is a negative transcriptional regulator of alpha1-adrenergic induced gene expression in cultured cardiac myocytes. AB - We recently reported that interleukin-1beta (IL-1beta) induces a novel form of cardiac myocyte hypertrophy characterized by an increase in protein content but an absence of the fetal program of skeletal alpha-actin or beta-myosin heavy chain (beta-MHC) gene expression (Palmer, J. N., Hartogensis, W. E., Patten, M., Fortuin, F. D., and Long, C. S. (1995) J. Clin. Invest. 95, 2555-2564). Because of the apparent disparity between this myocardial phenotype and that seen with other hypertrophic agents in culture, such as catecholamines, we investigated the effect of IL-1beta on alpha1-induced cardiomyocyte hypertrophy. Although there was no augmentation in total protein when IL-1beta and phenylephrine were given simultaneously, IL-1beta attenuated the increase in contractile protein mRNAs (skeletal alpha-actin and beta-MHC) in response to phenylephrine. Transient transfection studies with skeletal alpha-actin and beta-MHC promoter constructs linked to the chloramphenicol acetyltransferase (CAT)-reporter gene indicate that repression occurred at the level of gene transcription. In view of the previously reported activity of the zinc finger protein YY1 in the negative regulation of the skeletal alpha-actin promoter in cardiomyocytes (MacLellan, W. R., Lee, T. C., Schwartz, R. J., and Schneider, M. D. (1994) J. Biol. Chem. 269, 16754 16760), we investigated the potential role of this factor in the IL-1beta mediated effects. Using transient transfection, we found that a mutation in the YY1 binding site of the skeletal alpha-actin promoter abolished the inhibitory effect of IL-1beta. We further found that the 127-base pair fragment of the skeletal alpha-actin promoter required for the IL-1beta effect is also required for inhibition by the overexpression of YY1 in the myocytes. Furthermore, increased levels of YY1 protein are found in IL-1beta treated myocytes. Taken together these results suggest that the repression of contractile protein gene transcription by IL-1beta may be due, at least in part, to activation of the negative transcription factor YY1. PMID- 8702884 TI - The beta/alpha peak height ratio of ATP. A measure of free [Mg2+] using 31P NMR. AB - From 31P NMR measurements made in vitro at 38 degrees C, I = 0.25, pH 5. 75-8.5, and calculated free [Mg2+] from 0 to 5 mM, we show that, within the physiological range of cytosolic free [Mg2+] from 0.25 to 1.5 mM, the chemical shift difference between the alpha- and beta-ATP resonances, deltaalphabeta, changes by only 0.6 ppm. Consequently, we developed new formalisms from known acid and Mg2+ dissociation constants by which the observed chemical shift of Pi, deltaPi, and the peak height ratio of the beta- and alpha-ATP resonances, hbeta/alpha, could be related to free [Mg2+] by simultaneous solution of: [equation: see text] We found that hbeta/alpha changed 2.5-fold as free [Mg2+] varied from 0.25 to 1.5 mM, providing a more sensitive and accurate measure of free cytosolic [Mg2+]. In working rat heart perfused with glucose, free [Mg2+] was 1.0 +/- 0.1 from hbeta/alpha and 1.2 +/- 0.03 from measured [citrate]/[isocitrate] but 0.51 +/- 0.1 from deltaalphabeta. Addition of ketone bodies to the perfusate decreased free [Mg2+] estimated from hbeta/alpha to 0.61 +/- 0.02 and 0.74 +/- 0.11 by [citrate]/[isocitrate] but the estimate from deltaalphabeta was unchanged at 0.46 +/- 0.04 mM. Such differences in estimated free [Mg2+] alter the apparent Keq of the creatine kinase reaction and hence the estimated cytosolic free [SigmaADP]. PMID- 8702885 TI - Human tumor necrosis factor receptor p75/80 (CD120b) gene structure and promoter characterization. AB - Tumor necrosis factor receptor p75 (TNF-R p75) is a 75-kDa type I transmembrane protein expressed predominantly on cells of hematopoietic lineage. TNF-R p75 belongs to the TNF receptor superfamily characterized by cysteine-rich extracellular regions composed of three to six disulfide-linked domains. In the present report we have characterized, for the first time, the complete gene structure for human TNF-R p75, which spans approximately 43 kbp. The gene consists of 10 exons (ranging from 34 base pairs to 2.5 kilobase pairs) and nine introns (343 base pairs to 19 kilobase pairs). Consensus elements for transcription factors involved in T cell development and activation were noted in the 5'-flanking region including T cell factor-1, Ikaros, AP-1, CK-2, interleukin 6 receptor E (IL-6RE), ISRE, GAS, NF-kappaB, and Sp1. The unusual (GATA)n and (GAA)(GGA) repeats found within intron 1 may prove useful for further genome analysis within the 1p36 chromosomal locus. Characterization of the human TNF-R p75 gene structure will permit further assessment of its involvement in normal hematopoietic cell development and function, autoimmune disease, and nonrandom translocations in hematopoietic malignancies. PMID- 8702886 TI - The major subunit of the asialoglycoprotein receptor is expressed on the hepatocellular surface in mice lacking the minor receptor subunit. AB - The mammalian asialoglycoprotein receptor (ASGPR) is located on the sinusoidal membrane of hepatocytes where it binds and endocytoses galactose-terminated glycoproteins (asialoglycoproteins). ASGPR is composed of two highly homologous subunits, termed hepatic lectin 1 and 2. Despite numerous studies the contribution of both subunits to biosynthesis and functional activity of ASGPR in vivo has remained controversial. Mice lacking the murine hepatic lectin (MHL)-2 subunit are viable and fertile without obvious phenotypic abnormalities. In the absence of MHL-2, knockout mice express MHL-1 protein at reduced levels. Here, we examine the intracellular fate and function of this remaining subunit. The results show that MHL-1 reaches the hepatocellular surface in knockout mice but is unable to effectively remove any one of three different radiolabeled ligands within 30 min. A small but detectable residual ligand clearance in knockout mice at 4 h is apparently not mediated by remaining MHL-1. Serum concentrations of galactose-terminating glycoproteins are not elevated in these ASGPR-deficient mice. However, competitive in vitro degradation experiments suggest that other endogenous ASGPR ligands, the nature of which remain to be determined, accumulate in serum of knockout animals. PMID- 8702887 TI - Oxidative damage to DNA constituents by iron-mediated fenton reactions. The deoxycytidine family. AB - Damage by iron-mediated Fenton reactions under aerobic or anaerobic conditions to deoxycytidine, deoxycytidine-5'-monophosphate, d-CpC, d-CpCpC, and dCMP residues in DNA resulted in at least 26 distinguishable products. Of these, 24 were identified by high performance liquid chromatography retention times, radiolabeling, UV absorption spectra, chemical synthesis, fast atom bombardment mass spectrometry, high resolution fast atom bombardment mass spectrometry, and/or NMR. The nature of the products was qualitatively similar for each substrate except for d-CpC (and possibly d-CpCpC) under anaerobic conditions for which 5-hydroxy-deoxycytidine was uniquely present and 1-carbamoyl-1-carboxy-4-(2 deoxy-beta-D-erythropentofuranosyl) glycinamide was uniquely absent. Damage to dC, d-CpC, and d-CpCpC but not to dCMP or DNA was largely quenched by ethanol, indicating that iron is strongly associated only with dCMP and DNA. The presence of oxygen had little effect with dC or dCMP but had quantitative and qualitative effects with d-CpC and a significantly quantitative but not a qualitative effect with DNA. NADH could drive the Fenton reaction to cause damage to the dC family in vitro, consistent with a previous proposal that NADH was the reducing agent for the Fenton reaction in vivo (Imlay, J.A., and Linn, S. (1988) Science 240, 1302-1309). Finally, the damage spectrum of the dC family by the Fenton reaction is compared with that by ionizing radiation and chemical mechanisms leading to the formation of the 24 identified products are proposed. PMID- 8702888 TI - Oxidative damage to DNA constituents by iron-mediated fenton reactions. The deoxyguanosine family. AB - 2'-Deoxyguanosine, 3'-dGMP, 5'-dGMP, d-GpG, or double-stranded DNA were exposed to H2O2 in the presence of Fe2+ under anaerobic conditions or under aerobic conditions in the presence of Fe3+, Fe2+, Fe2+/NADH, or Fe3+/NADH with and without ethanol. The products were enzymatically digested to nucleosides, separated by high performance liquid chromatography (HPLC), quantified, and characterized by HPLC retention time, radiolabeling, UV absorbance spectrometry, NMR, and mass spectrometry. 20 products, constituting 78-81% of the damage, were distinguished from aerobic reactions of Fe2+/H2O2 with dG and dGMP, 16 of which were identified. The product spectra from dG, 3'-dGMP, and 5'-dGMP differ from one another, and the spectrum of the 5' nucleoside of d-GpG differs from that of the 3' nucleoside. 7, 8-Dihydro-8-oxo-2'-deoxyguanosine is the most abundant DNA bound product aside from abasic sites, and its formation was more closely analyzed. Both NADH, which can reduce Fe3+, and ethanol, which can scavenge some free radicals, change the product profiles. Damage enhancement by NADH follows the sequence dG < d-GpG < 3'-dGMP < 5'-dGMP < DNA; the reverse sequence is observed for ethanol quenching. This sequence of susceptibility and the product differences seen for the 3' and 5' phosphate may reflect localization of iron and the damaging radicals upon the substrate. PMID- 8702889 TI - The role of carboxyl-terminal basic amino acids in Gqalpha-dependent activation, particulate association, and nuclear localization of phospholipase C-beta1. AB - The phospholipase C (PLC)-beta isozymes differ from the PLC-gamma and PLC-delta isozymes in that they possess a long COOH-terminal sequence downstream of their catalytic domain, are activated by alpha subunits of the Gq class of G proteins, associate with the particulate subcellular fraction, and are present in the nucleus. Most of the COOH-terminal domain of PLC-beta isozymes is predicted to be helical, and three regions in this domain, PLC-beta1 residues 911-928 (region 1), 1055-1072 (region 2), and 1109-1126 (region 3), contain a high proportion of basic residues that are highly conserved. Projection of the sequences of these three regions in helical wheels reveals clustering of the basic residues. The role of the COOH terminus and the clustered basic residues in PLC-beta1 was investigated by either truncating the entire COOH-terminal domain (mutant DeltaC) or replacing two or three clustered basic residues with isoleucine (or methionine), and expressing the mutant enzymes in CV-1, Rat-2, or Swiss 3T3 cells. The DeltaC mutant no longer showed the ability to be activated by Gqalpha, to translocate to the nucleus, or to associate with the particulate fraction. Substitution of clusters of basic residues in regions 1 and 2 generally reduced the extent of activation by Gqalpha, whereas substitution of a basic cluster in region 3 had no effect. Substitution of the cluster of lysine residues 914, 921, and 925 in region 1 had the most marked effect, reducing Gqalpha-dependent activity to 10% of that of wild type. All substitution mutants, with the exception of that in which lysine residues 1056, 1063, and 1070 in region 2 were substituted with isoleucine, behaved like the wild-type enzyme in showing an approximately equal distribution between cytoplasm and nucleus; only 12% of the region 2 mutant was present in the nucleus. None of the basic clusters appeared critical for particulate association; however, replacement of each cluster reduced the amount of PLC-beta1 in the particulate fraction by some extent, suggesting that all the basic residues contribute to the association, presumably by interacting with acidic residues in the particulate fraction. Membrane localization of PLC-beta isozymes is therefore likely mediated by both the COOH terminal domain and the pleckstrin homology domain, the latter of which is known to bind phosphatidylinositol 4,5-biphosphate. PMID- 8702890 TI - Identification of residues in the translocation pathway of EmrE, a multidrug antiporter from Escherichia coli. AB - EmrE is a small, 12-kDa, highly polyspecific antiporter, which exchanges hydrogen ions with aromatic cations such as methyl viologen. EmrE-mediated transport is inhibited by the sulfhydryl-reactive reagent 4-(chloromercuri)benzoic acid (PCMB) but not by a variety of other sulfhydryl reagents. This differential effect is due to the fact that the organic mercurial is a substrate of the transporter and can reach domains otherwise inaccessible to the different reagents. To find out which of the three cysteine residues in EmrE is reacting with PCMB, each was replaced with serine and it was shown that none of them is essential for transport activity. A protein completely devoid of Cys residues (CL) is also capable of substrate accumulation albeit at a slower rate. Mutated proteins in which only one of the native cysteines was left whereas the other changed to serine were also constructed. The use of these proteins demonstrated that two of the three Cys in EmrE, Cys-41 and Cys-95, but not Cys-39, react with PCMB. A related mercurial, 4-(chloromercuri)benzenesulfonic acid (PCMBS), is only a very poor inhibitor, probably because of the negative charge it bears. PCMBS reacts with EmrE in an asymmetric and unique way. It reacts with the mutant bearing a single Cys residue in position 95 (CL-C95) only when the reagent is present in the outside face of the membrane and with the mutant CL-C41 only when allowed to permeate to the cell interior; as expected, it does not react with the mutant protein bearing a single Cys at position 39 (CL-C39). It is concluded that PCMB permeates through the substrate pathway of EmrE and covalently reacts with the two exposed residues, Cys-95 and Cys-41, but not with Cys-39, located on the opposite face of the helix relative to residue 41. In addition, because of the asymmetric reactivity to PCMBS, an inhibitor that does not permeate through the protein, it is concluded that Cys-41 is closer to the cytoplasmic face than Cys 95. The results demonstrate the existence of a domain accessible only to substrates and provide a unique tool for studying the substrate permeation pathway of an ion-coupled transporter. PMID- 8702891 TI - Expression of purinergic receptor channels and their role in calcium signaling and hormone release in pituitary gonadotrophs. Integration of P2 channels in plasma membrane- and endoplasmic reticulum-derived calcium oscillations. AB - The role of ATP as a positive feedback element in Ca2+ signaling and secretion was examined in female rat pituitary gonadotrophs. ATP and ADP, but not AMP or adenosine, induced a dose- and extracellular Ca2+-dependent rise in [Ca2+]i in identified gonadotrophs in a Mg2+- and suramin-sensitive manner. ATP, adenosine 5'-O-(3-thiotriphosphate), adenosine-5'-O-(1-thiotriphosphate), 2-methylthio-ATP, and 3'-O-(4-benzoyl)benzoyl-ATP were roughly equipotent in rising [Ca2+]i in gonadotrophs, while ADP was effective only at submillimolar concentration range, and none of these compounds permeabilized the cells. On the other hand, alpha,beta-methylene-ATP, beta,gamma-methylene-ATP, and UTP were unable to induce any rise in [Ca2+]i. This pharmacological profile is consistent with expression of P2X2 and/or P2X5 purinergic receptor channels. Patch-clamp experiments showed that ATP induced an inward depolarizing current in gonadotrophs clamped at -90 mV, associated with an increase in [Ca2+]i. The ATP-induced [Ca2+]i response was partially inhibited by nifedipine, a blocker of voltage-sensitive Ca2+ channels (VSCC), but was not affected by tetrodotoxin, a blocker of voltage-sensitive Na+ channels. Thus, the P2-depolarizing current itself drives Ca2+ into the cell, but also activates Ca2+ entry through VSCC. In accord with this, low [ATP] induced plasma membrane-dependent [Ca2+]i oscillations in quiescent cells, and increased the frequency of spiking in spontaneously active cells. ATP-induced Ca2+ influx also affected agonist-induced and InsP3-dependent [Ca2+]i oscillations by increasing the frequency, base line, and duration of Ca2+ spiking. In addition, ATP stimulated gonadotropin secretion and enhanced agonist-induced gonadotropin release. ATP was found to be secreted by pituitary cells during agonist stimulation and was promptly degraded by ectonucleotidase to adenosine. These observations indicate that ATP represents a paracrine/autocrine factor in the regulation of Ca2+ signaling and secretion in gonadotrophs, and that these actions are mediated by P2 receptor channels. PMID- 8702892 TI - Imparting exquisite specificity to peanut agglutinin for the tumor-associated Thomsen-Friedenreich antigen by redesign of its combining site. AB - Lectins from legumes constitute one of the most thoroughly studied families of proteins, yet the absence of a rigorous framework to explain their carbohydrate binding specificities appears to have prevented a rational approach to alter their ligand binding activity. Studies reported here deal with the redesign of the recognition propensity of peanut agglutinin (PNA), an important member of the family. PNA is extensively used as a tool for recognition of the tumor-associated Thomsen-Friedenrich antigen (T-antigen; Galbeta1-3GalNAc) on the surfaces of malignant cells and immature thymocytes. PNA also recognizes N-acetyllactosamine (LacNAc; Galbeta1-4GlcNAc), which is present at the termini of several cell surface glycoproteins. The crystal structure of the PNA-lactose complex revealed, in addition to the expected interactions with the residues constituting the binding site, the presence of leucine 212 at a position close enough to be in steric contact with the acetamido group on LacNAc. We report here two leucine mutants, one to asparagine (L212N) and the other to alanine (L212A), that exhibit distinct preference for T-antigen and N-acetyllactosamine, respectively. Carbohydrate binding studies reveal that mutant L212N does not recognize LacNAc at high concentrations, thus making it an exquisitely specific cell-surface marker compared with its wild-type counterpart. PMID- 8702893 TI - Redirected cytotoxic effector function. Requirements for expression of chimeric single chain high affinity immunoglobulin e receptors. AB - The aim of this study was to construct a single chain chimeric FcepsilonRIalpha receptor capable of effector function in leukocytes, including cytotoxic lymphocytes. To determine the most effective single chain FcepsilonRIalpha receptor with respect to IgE binding and signaling function, a variety of chimeric gene constructs were transiently transfected into COS-7 cells. The most effective chimera consisted of four parts including: wild-type or mutated extracellular domains (Trp130 --> Ala130, W130A) of FcepsilonRIalpha, membrane proximal and transmembrane regions of FcgammaRIIa, and intracellular CD3zeta (epsilonIIaIIazeta). Scatchard analysis indicated that these FcepsilonRIalpha chimeric receptor bound ligand with an affinity of 0.9 to 2.2 x 10(9) -1. Ligand binding capacity was dramatically reduced with the deletion of 11 membrane proximal amino acids of FcepsilonRIalpha; however, function was restored by substitution with the equivalent region of FcgammaRIIa, suggesting a crucial requirement for a "spacer" segment between the transmembrane and extracellular ligand binding domain. Chimeras that bound IgE effectively also mediated phagocytosis. Chimeric receptors that contained transmembrane zeta were expressed as multimers and consequently did not bind IgE effectively; however, cotransfection of these chimeras with gamma-chain largely reconstituted IgE mediated phagocytosis. The mouse cytotoxic T lymphocyte cell line, CTLLR8 was stably transfected with epsilonIIaIIazeta, and cloned transfectants were demonstrated to lyse target cells in an anti-FcepsilonRIalpha or IgE antibody dependent manner. Therefore, functional single chain chimeric FcepsilonRIalpha receptors were expressed in the absence or presence of associated zeta or gamma molecules and were used to redirect killer lymphocytes to target cells. PMID- 8702894 TI - High affinity binding of ankyrin induced by volume expansion in skate erythrocytes. AB - Volume expansion of little skate (Raja erinacea) erythrocytes increases the affinity of ankyrin binding without altering in the number of binding sites. Potassium iodide-stripped inside-out vesicles (KI-IOV) were used to assess ankyrin binding under volume-expanded conditions. Under isoosmotic conditions, ankyrin binds nearly exclusively to a single class of sites (Bmax, 52 +/- 12 microg/mg; Kd, 150 +/- 28 nM). KI-IOV from volume-expanded cells (either with one half osmolarity medium or with inclusion of the permeant solute ethylene glycol) demonstrate two ankyrin-binding populations. A high affinity population occurs transiently under volume-expanded conditions. This population has a Bmax of 18 +/ 7 microg/mg and a Kd of 25 +/- 9 nM. Total binding of high and low affinity sites is 57 +/- 17 microg/mg. This change in ankyrin affinity is reversible on volume regulatory decrease. A major target protein in the KI-IOV was identified as the skate homolog of the mammalian red cell anion exchanger band 3. Inclusion of the purified cytoplasmic domain of band 3 competes away more than 80% of the ankyrin binding. To determine whether increased ankyrin affinity is due to band 3 tetramer formation that occurs in volume expansion, cells were treated with pyridoxal 5-phosphate or 4,4'-dinitrostilbene-2,2'-disulfonic acid, two agents that increase tetramer formation under isoosmotic conditions. Both treatments altered the binding affinity with a shift toward higher affinity binding without significant alteration in the number of binding sites. PMID- 8702895 TI - Influence of the mature portion of a precursor protein on the mitochondrial signal sequence. AB - Most mitochondrial proteins are synthesized with an N-terminal signal sequence that targets these proteins to various compartments within the mitochondria. Signal sequences have been shown to be functional by fusing them to a nonmitochondrial passenger protein and observing import. In many cases, a signal sequence has been fused to passenger proteins, such as dihydrofolate reductase, and import occurred. There are, though, several unexplained instances in which a signal sequence was attached to a passenger protein and import was not observed. In this study, the N-terminal 23 residues of the matrix enzyme rhodanese could import several passenger proteins but were unable to import the mature form of mitochondrial aldehyde dehydrogenase (mALDH). However, if these same 23 residues were fused to the middle portion of mALDH, import was recovered, suggesting that the rhodanese signal sequence and N terminus of mALDH were incompatible for import. Circular dichroism data indicated that a peptide corresponding to the region of fusion between rhodanese and mALDH had less structure than corresponding peptides from imported fusion proteins, suggesting that mALDH may alter the helix in the rhodanese signal sequence, thus preventing import. PMID- 8702896 TI - Biochemical and biophysical studies of reactive center cleaved plasminogen activator inhibitor type 1. The distance between P3 and P1' determined by donor donor fluorescence energy transfer. AB - Plasminogen activator inhibitor type 1 (PAI-1) is a fast acting inhibitor of plasminogen activators (PAs). In accordance with other serpins, PAI-1 is thought to undergo a conformational change upon reactive center cleavage. In this study we have developed methods to produce and purify reactive center cleaved wild-type PAI-1 and characterized this molecular form of PAI-1 by biochemical and biophysical methods. Incubation with Sepharose-bound trypsin caused cleavage only at the P1-P1' bond in the reactive center and resulted in 39- and 4-kDa polypeptides, strongly held together by noncovalent interactions. Circular dichroism measurements suggest that the reactive center cleavage triggers larger conformational changes than the conversion from the active to the latent form. Cleaved PAI-1 did not bind to either PAs or vitronectin but retained the heparin binding capacity. To study the structure of cleaved PAI-1 by polarized fluorescence spectroscopy and to measure intramolecular distances, we used cysteine substitution mutants to which extrinsic fluorescence probes were attached. These studies revealed increasing orientational freedom of probes in the P3 and P1' positions upon cleavage. Distance measurements based on fluorescence energy transfer between probes in positions P3 and P1' indicate that these residues are separated by at least 68 +/- 10 A in cleaved PAI-1. PMID- 8702897 TI - The role of specific lysine residues in the passage of anions through the Pseudomonas aeruginosa porin OprP. AB - When grown under phosphate-limiting conditions Pseudomonas aeruginosa expresses the phosphate-specific porin OprP. In order to determine whether any of the lysine residues located in the amino-terminal half of the protein play a role in the transport of anions through the channels, the first nine amino-terminal lysine residues of OprP were substituted with glutamates. The mutant proteins were purified and the channels they formed were characterized by reconstituting the purified porins in planar lipid membranes. In comparison to the wild-type protein, the Lys74, Lys121, and Lys126 mutants all displayed reduced levels of conductance at KCl concentrations below 1 M, and the Lys74 and Lys121 mutants no longer exhibited a saturation of conductance at high anion concentrations. In addition, the ability of phosphate ions to inhibit the conductance of Cl- ions through the channels formed by the Lys121 mutant was greatly reduced, while their ability to inhibit the Cl- conductance of the Lys74 mutant was reduced by approximately 2-fold. To clarify the roles that Lys74, Lys121, and Lys126 play in regulating the channel characteristics of OprP, these amino acids were replaced with either glycine or glutamine residues. Analysis of these mutants suggested that both Lys74 and Lys126 may serve to funnel anions toward the binding site, but only the presence of Lys121 is required for the formation of the inorganic phosphate-specific binding site of OprP. PMID- 8702898 TI - Ligand-dependent conformational plasticity of the periplasmic histidine-binding protein HisJ. Involvement in transport specificity. AB - The periplasmic histidine permease of Salmonella typhimurium is composed of a membrane-bound complex and a soluble histidine-binding protein (the periplasmic receptor), HisJ. Liganded receptor interacts with the membrane-bound complex, inducing ATP hydrolysis and substrate translocation. Preliminary evidence had shown a lack of direct correlation between the affinity of HisJ for a ligand and translocation efficiency, suggesting that the precise form of the receptor is important in determining its interaction with the membrane-bound complex. We have investigated the nature of the conformations assumed by HisJ upon binding a variety of ligands by tryptophan fluorescence enhancement, reaction with a closed form-specific monoclonal antibody, and changes in UV absorption spectra. It is demonstrated that although HisJ binds all the ligands and undergoes a conformational change, it assumes measurably different conformations. We also show that the interaction between HisJ and the membrane-bound complex depends on the nature of the ligand. Transport specificity appears to be defined, at least in part, by the conformation of the bound receptor, manifested either by the effect of a given ligand on the closed structure per se, or by the effect of ligand association on the equilibrium constant relating the open and the closed liganded forms. PMID- 8702899 TI - Beta-galactosidase enzymatic activity as a molecular probe to detect specific antibodies. AB - The main antigenic region of foot-and-mouth disease virus serotype C1, also called site A, has been inserted in zones of the beta-galactosidase important for the stabilization of the active site, causing important changes in the Km and the specific activity of the resulting enzymes. The peptide is displayed at the surface of the recombinant proteins and, in all the cases, presents a good antigenicity. Among the recombinant proteins constructed, in proteins M278VP1 and M275SVP1 the peptide is inserted in a large loop of the beta-galactosidase (amino acids 272-288) involved in the formation of the activating interface. In these constructs, the binding of the specific antibodies directed to the foreign peptide causes an increase of the beta-galactosidase activity up to about 200%. This phenomenon has been proved using monoclonal antibodies and also using polyclonal sera generated against the peptide. Different hypothesis of the mechanism of modulation upon antibody binding are discussed. This insertion site seems to be sensitive enough to enzymatic modulation mediated by antibody binding. We propose further exploring this insertion site as a tool for a rapid detection of specific antibodies in a quick and simple homogeneous assay based on the colorimetric determination of beta-galactosidase activity. PMID- 8702900 TI - Dual reactivity of several monoclonal anti-nucleosome autoantibodies for double stranded DNA and a short segment of histone H3. AB - We have shown previously that four IgG monoclonal autoantibodies (mAbs) reacted in ELISA with both double-stranded (ds) DNA and peptide 83-100 of histone H3. The peptide 83-100 contains a cysteine residue at position 96 and readily dimerizes at pH 7-8. We describe here that only the 83-100 dimers, and not the 83-100 monomers, are recognized by the four antibodies and inhibit in ELISA the binding of mAbs to dsDNA. The equilibrium affinity constants (Ka) and kinetic rate constants of two of these mAbs were measured in a biosensor system. Ka values were significantly higher when these mAbs were tested with dsDNA as compared with the 83-100 dimer. Further higher Ka values were measured with mononucleosomes containing DNA and histones. It is proposed that these four mAbs are directed against a topographic determinant formed by DNA and the region 83-100 of H3 present as a dimer at the surface of nucleosome, and that they react, although significantly less well, with DNA and peptide dimer tested separately. This study provides a quantitative and kinetic basis to interaction between several antibodies and distinct antigenic structures and allows us to better understand the structural basis of apparent autoantibody cross-reactivity. PMID- 8702902 TI - Characterization of a double cellulose-binding domain. Synergistic high affinity binding to crystalline cellulose. AB - Most cellulose-degrading enzymes have a two-domain structure that consists of a catalytic and a cellulose-binding domain (CBD) connected by a linker region. The linkage and the interactions of the two domains represent one of the key questions for the understanding of the function of these enzymes. The CBDs of fungal cellulases are small peptides folding into a rigid, disulfide-stabilized structure that has a distinct cellulose binding face. Here we describe properties of a recombinant double CBD, constructed by fusing the CBDs of two Trichoderma reesei cellobiohydrolases via a linker peptide similar to the natural cellulase linkers. After expression in Escherichia coli, the protein was purified from the culture medium by reversed phase chromatography and the individual domains obtained by trypsin digestion. Binding of the double CBD and its single CBD components was investigated on different types of cellulose substrates as well as chitin. Under saturating conditions, nearly 20 micromol/g of the double CBD was bound onto microcrystalline cellulose. The double CBD exhibited much higher affinity on cellulose than either of the single CBDs, indicating an interplay between the two components. A two-step model is proposed to explain the binding behavior of the double CBD. A similar interplay between the domains in the native enzyme is suggested for its binding to cellulase. PMID- 8702901 TI - Induction of phosphoglycerate kinase 1 gene expression by hypoxia. Roles of Arnt and HIF1alpha. AB - To identify new dimerization partners for the aromatic hydrocarbon receptor nuclear translocator (Arnt), we used its N-terminal region (amino acids 1-470) as a target in a two-hybrid screening procedure, and we cloned the murine form of hypoxia-inducible factor 1alpha (HIF1alpha). Sequence comparisons reveal substantial identity between mouse and human HIF1alpha. Hypoxia induces a 10-fold accumulation of phosphoglycerate kinase 1 mRNA in wild type mouse hepatoma (Hepa 1c1c7) cells; the induction mechanism is Arnt dependent because induction does not occur in Arnt-defective cells. Furthermore, induction of phosphoglycerate kinase 1 mRNA requires Arnt's N-terminal region, which mediates DNA binding and heterodimerization; in contrast, induction does not require Arnt's C-terminal region, which mediates transactivation. We also show that a GAL4-HIF1alpha fusion protein transactivates a GAL4-dependent gene in the absence of Arnt, that HIF1alpha's transactivation capability is inducible by hypoxia, and that both hypoxia responsiveness and transactivation capability reside within the C terminal 83 amino acids of HIF1alpha. Our findings generate new insights into the mechanism by which Arnt and HIF1alpha induce transcription in response to hypoxia. PMID- 8702903 TI - Effect of cAMP binding site mutations on the interaction of cAMP receptor protein with cyclic nucleoside monophosphate ligands and DNA. AB - Although cAMP binding to wild type cAMP receptor protein (CRP) induces specific DNA binding and activates transcription, cyclic nucleoside monophosphate (cNMP) binding to the CRP mutant Ser128 --> Ala does not, whereas the double CRP mutant Thr127 --> Leu/Ser128 --> Ala activates transcription even in the absence of cNMP. Isothermal titration calorimetry measurements on the cNMP binding reactions to the S128A and T127L/S128A mutants show that the reactions are mainly entropically driven as is cAMP binding to CRP. In contrast to cAMP binding to CRP, the binding reactions are noncooperative and exothermic with binding enthalpies (DeltaHb) ranging from -23.4 +/- 0.9 kJ mol-1 for cAMP binding to S128A at 39 degrees C to -4.1 +/- 0.6 kJ mol-1 for cAMP binding to T127L/S128A at 24 degrees C and exhibit enthalpy-entropy compensation. To account for the inactivity of the S128A mutant, in vitro and in vivo DNA binding experiments were performed on the cAMP-ligated S128A mutant. The cAMP-ligated S128A mutant binds to the consensus DNA binding site with approximately the same affinity as that of cAMP-ligated CRP but forms a different type of complex, which may account for loss of transcriptional activity by the mutant. Energy minimization computations on the cAMP-ligated S128A mutant show that amino acid conformational differences between S128A and CRP occur at Ser179, Glu181, and Thr182 in the center of the DNA binding site, implying that these conformational changes may account for the difference in DNA binding. PMID- 8702904 TI - Effect of cystic fibrosis-associated mutations in the fourth intracellular loop of cystic fibrosis transmembrane conductance regulator. AB - The cystic fibrosis transmembrane conductance regulator (CFTR) contains multiple membrane spanning sequences that form a Cl- channel pore and cytosolic domains that control the opening and closing of the channel. The fourth intracellular loop (ICL4), which connects the tenth and eleventh transmembrane spans, has a primary sequence that is highly conserved across species, is the site of a preserved sequence motif in the ABC transporter family, and contains a relatively large number of missense mutations associated with cystic fibrosis (CF). To investigate the role of ICL4 in CFTR function and to learn how CF mutations in this region disrupt function, we studied several CF-associated ICL4 mutants. We found that most ICL4 mutants disrupted the biosynthetic processing of CFTR, although not as severely as the most common DeltaF508 mutation. The mutations had no discernible effect on the channel's pore properties; but some altered gating behavior, the response to increasing concentrations of ATP, and stimulation in response to pyrophosphate. These effects on activity were similar to those observed with mutations in the nucleotide-binding domains, suggesting that ICL4 might help couple activity of the nucleotide-binding domains to gating of the Cl- channel pore. The data also explain how these mutations cause a loss of CFTR function and suggest that some patients with mutations in ICL4 may have a milder clinical phenotype because they retain partial activity of CFTR at the cell membrane. PMID- 8702905 TI - Sites of ribosomal RNAs involved in the subunit association of tight and loose couple ribosomes. AB - It was demonstrated previously by kethoxal treatment studies that tight (TC) and loose (LC) couple ribosomes use different sites of the 16 S and 23 S RNAs for subunit association (Burma, D. P., Srivastava, A. K., Srivastava, S., and Dash, D. (1985) J. Biol. Chem. 260, 10517-10525). To localize these sites, a number of oligodeoxynucleotides complementary to the suspected sites of the 16 S and 23 S RNAs were synthesized, and their binding to ribosomes and effects on subunit association were studied. Some of the probes inhibit the association of both TC and LC ribosomes, some inhibit the association of TC but not LC ribosomes, and some inhibit the association of LC but not TC ribosomes. It appears that both TC and LC ribosomes use one common site of association, bases 818-823 of the 16 S and 2308-2313 of the 23 S RNA. The second site, 788-793 of the 16 S RNA and 2753 2758 of the 23 S RNA for TC ribosomes, and 783-791 of the 16 S RNA and 2295-2303 of the 23 S RNA for LC ribosomes, is not shared. This suggests a spatial movement of the ribosomal subunits with respect to one another and lends further support to the model of translocation based on the interconversion of TC and LC ribosomes. PMID- 8702906 TI - G proteins regulate dihydropyridine binding to moss plasma membranes. AB - The role of calcium as an activator and regulator of many biological processes is linked to the ability of the cell to rapidly change its cytoplasmic calcium levels. Calcium acts as an intracellular messenger in hormone-induced bud formation during the development of the moss Physcomitrella patens. Calcium transport and ligand binding studies have implicated plasma membrane-localized 1, 4-dihydropyridine (DHP)-sensitive calcium channels in this increase in cellular calcium. To understand the regulation of the moss calcium channel, we investigated the involvement of GTP binding regulatory proteins (G proteins). Guanosine 5'-(gamma-thio)triphosphate (GTPgammaS), a nonhydrolyzable GTP analog that locks G proteins into their active state, stimulated DHP binding to high affinity receptors in the moss plasma membrane. DHP binding was measured as the ability of the DHP agonist Bay K8644 or the DHP antagonist nifedipine to compete with the DHP arylazide [3H]azidopine for binding to moss plasma membranes. G protein stimulation of binding was seen when competition was carried out with either nifedipine or Bay K8644. G proteins regulated the rates of association and dissociation of bound [3H]azidopine, and stimulation was dependent on GTPgammaS concentration. Guanosine 5'-(beta-thio)diphosphate, a GDP analog that locks G proteins into their inactivated state, did not affect the dose dependence of either the agonist or the antagonist. These results suggest that G proteins may act via a membrane-delimited pathway to regulate calcium channels in the moss plasma membrane. PMID- 8702907 TI - A GC-rich region containing Sp1 and Sp1-like binding sites is a crucial regulatory motif for fatty acid synthase gene promoter activity in adipocytes. Implication In the overactivity of FAS promoter in obese Zucker rats. AB - We have previously shown that the proximal 2-kb sequence of the fatty acid synthase (FAS) promoter transfected into rat adipocytes was highly sensitive to the cellular context, displaying an overactivity in obese (fa/fa) versus lean Zucker rat adipocytes. Using deletional analysis, we show here that FAS promoter activity mainly depends on a region from -200 to -126. This sequence exerts a strong negative effect on FAS promoter in adipocytes from lean rats but not in those from obese rats, resulting in a marked overtranscriptional activity in the latter cells. This region, fused to a heterologous promoter, the E1b TATA box, induced differential levels of gene reporter activity in lean and obese rat adipocytes, indicating it harbors fa-responsive element(s). Whatever the rat genotype, adipocyte nuclear proteins were shown to footprint the same protected sequence within the fa-responsive region, and supershift analysis demonstrated that Sp1 or Sp1-like proteins were bound to this DNA subregion. Compelling evidence that the Sp1 binding site contained in this sequence was implicated in the differential promoter activity in lean versus obese rats, was provided by the observation that mutations at this Sp1 site induced a 2.5-fold increase in FAS promoter activity in adipocytes from lean rats, whereas they had no effect in adipocytes from obese rats. PMID- 8702908 TI - Regulation of gene expression of a binding protein for fibroblast growth factors by retinoic acid. AB - Retinoids are potent regulators of growth and differentiation and have shown promise as chemotherapeutic agents against selected cancers in particular squamous cell carcinoma (SCC). Earlier studies from our laboratory showed that a secreted binding protein for fibroblast growth factors (BP) is expressed at high levels in SCC cell lines and tissue samples. Here we investigate whether retinoids affect BP gene expression in SCC. In six different human SCC cell lines, we found that all-trans-retinoic acid (tRA) down-regulated BP mRNA by 39 89% within 24 h. From this group of cell lines, we selected the ME-180 cell line for more detailed studies of the mechanisms of this regulation. tRA down regulated BP mRNA in a time- and dose-dependent manner. The effect of tRA was reversible, and BP mRNA returned to control levels within 24 h after removal of tRA. We also measured BP mRNA half-life and performed nuclear run-on experiments to study if tRA down-regulates BP by destabilizing the mRNA and/or by decreasing the rate of transcription. BP mRNA in ME-180 cells is very stable with a half life of >16 h, and tRA decreased BP mRNA with a half-time of 5 h. Actinomycin D and cycloheximide blocked the tRA effect, suggesting that transcriptional regulation as well as de novo protein synthesis contribute to this post transcriptional regulation of BP mRNA levels. In addition, tRA decreased the rate of BP gene transcription by 2- to 3-fold within 1 h. We conclude that retinoids down-regulate BP gene expression by post-transcriptional as well as by transcriptional mechanisms. PMID- 8702909 TI - Chronic opioid treatment induces adenylyl cyclase V superactivation. Involvement of Gbetagamma. AB - It has been known for some time that chronic treatment of neuronal cells and tissues with opioids, contrary to their acute effect, leads to an increase in cAMP accumulation. This phenomenon, defined as adenylyl cyclase superactivation, has been implicated in opiate addiction, yet the mechanism by which it is induced remains unclear. Here, we show that this phenomenon can be reproduced and studied in COS-7 cells cotransfected with adenylyl cyclase type V and mu-opioid receptor cDNAs. These cells display acute opioid inhibition of adenylyl cyclase activity, whereas prolonged exposure to the mu-agonist morphine or [-Ala2, N-methyl-Phe4, Gly-ol5]enkephalin leads to a time-dependent superactivation of adenylyl cyclase. This superactivated state is reversible, because it is gradually lost following agonist withdrawal. Adenylyl cyclase superactivation can be prevented by pertussis toxin pretreatment, indicating the involvement of Gi/o proteins, or by cotransfection with the carboxyl terminus of beta-adrenergic receptor kinase or with alpha-transducin (scavengers of Gbetagamma dimers), indicating a role for the G protein betagamma dimers in adenylyl cyclase superactivation. However, contrary to several other Gbetagamma-dependent signal transduction mechanisms (e.g. the extracellular signal-regulated kinase 2/MAP kinase pathway), adenylyl cyclase superactivation is not affected by the Ras dominant negative mutant N17 Ras. PMID- 8702910 TI - Distinct N-methyl-D-aspartate receptor 2B subunit gene sequences confer neural and developmental specific expression. AB - Expression of the N-methyl--aspartate (NMDA) receptor 2B (NR2B) subunit is neural specific and differentially regulated. It is expressed in the forebrain and in cerebellar granule cells at early postnatal stages and selectively repressed in the cerebellum after the second postnatal week, where it is replaced by the NR2C subunit. This switch confers distinct properties to the receptor. In order to understand the molecular mechanisms that differentially regulate the NR2B gene in the forebrain and cerebellum during development, we have isolated and characterized the promoter region of the NR2B gene. Two 5' noncoding exons and multiple transcription start sites were identified. Transcriptional analysis in transgenic mice reveals that an upstream 800-base pair region, which includes the first exon, is sufficient to direct neural-specific transcription. Developmental repression of the gene in the cerebellum requires additional regulatory elements residing in the first intron or second exon. Sequence elements that may participate in the regulation of the NR2B gene were identified by comparison to other neural genes. These studies provide insight into the molecular mechanisms regulating the switch of NMDA receptor subunit expression in the cerebellum, which ultimately account for the physiological changes in receptor function during development. PMID- 8702911 TI - Characterization of a novel member of the macrophage mannose receptor type C lectin family. AB - The recognition of a diversity of carbohydrates by the various calcium dependent (type C) lectin family members has been shown to be critical for a variety of processes ranging from cell adhesion to antigen presentation. Examination of the expressed sequence tag (EST) data base for novel type C lectins using E-selectin as a probe resulted in the identification of a distantly related short polypeptide sequence containing many of the conserved residues found in these carbohydrate-binding proteins. Cloning of the full-length murine cDNA containing this region revealed that this protein is a novel member of the family that includes the macrophage mannose, the phospholipase A2, and the DEC 205 receptors, with a cysteine-rich domain, a fibronectin type 2 domain, eight type C lectin domains, a transmembrane domain, and a short cytoplasmic carboxyl terminus. Genomic Southern analysis suggests that this is a conserved protein, and examination of a human homologue revealed a high degree of sequence homology with the murine form. Northern blot analysis revealed expression of a large transcript in a number of different human and murine tissues and tumor cells and an alternatively spliced smaller transcript with a divergent 5' sequence was expressed specifically in the human fetal liver. Analysis of the genomic structure revealed that the gene encoding this lectin was interrupted by a large number of introns, and the intron structure was similar to the macrophage mannose receptor gene. Finally, in situ hybridization analysis demonstrated that the transcript encoding this lectin was found in a number of highly endothelialized sites as well as in chondrocytes in cartilaginous regions of the embryo. PMID- 8702912 TI - c-Krox binds to several sites in the promoter of both mouse type I collagen genes. Structure/function study and developmental expression analysis. AB - We have previously shown that c-Krox is a zinc finger protein that can increases the transcriptional activity of the mouse alpha1(I) collagen promoter through its binding to two GC-rich sequences (Galera, P., Musso, M., Ducy, P., and Karsenty, G. (1994) Proc. Natl. Acad. Sci. U. S. A. 91, 9372-9376). In this report we show that c-Krox can bind to an additional site in the promoter of the alpha1(I) collagen gene and to three sites in the promoter of the alpha2(I) collagen gene, the other gene coding for type I collagen. One of the binding sites present in both promoters is adjacent to the CCAAT box. We have performed a structure/function analysis of c-Krox locating the transactivation domain in the zinc finger and C-terminal domains and the dimerization domain in the C-terminal end of the protein. We also demonstrate that c-Krox is an early response gene, whose expression is detectable as early as 9.5-day postcoitum in mouse embryos. Whole-mount in situ hybridization shows that c-Krox is expressed in dermatomes, the somite derivatives that generate dermis, and section in situ hybridization shows that c-Krox and alpha1(I) collagen mRNAs colocalized in skin but not in bone during development. This result is consistent with the predominant expression of c-Krox in skin in postnatal life. Thus, our findings suggest that c Krox is one transcription factor controlling the coordinated expression of the two type I collagen genes in skin. PMID- 8702913 TI - Primary folding of aspartylglucosaminidase. Significance of disulfide bridges and evidence of early multimerization. AB - Aspartylglucosaminidase (AGA) is a lysosomal enzyme involved in the degradation of N-linked glycoproteins in lysosomes. AGA is synthesized as an inactive precursor molecule, which is rapidly activated in the endoplasmic reticulum by a proteolytic cleavage into alpha- and beta-subunits. We have recently determined the three-dimensional structure of AGA and shown that it is a globular molecule with a heterotetrameric (alphabeta)2 structure. On the basis of structural and functional analyses, AGA seems to be the first mammalian protein belonging to a newly described protein family, the N-terminal nucleophile hydrolases. Because the activation of the prokaryotic members of the N-terminal nucleophile hydrolase family seems to be triggered by the assembly of the subunits, we have studied the initial folding and oligomerization of AGA and provide evidence that dimerization of two precursor molecules in the endoplasmic reticulum is a prerequisite for the activation of AGA. To gain further information on the structural determinants influencing the early folding of AGA, we used site-specific mutagenesis of cysteine residues to define the role of intrachain disulfide bridges in the folding and activation of the enzyme. The N-terminal disulfide bridges in both the alpha- and beta-subunits seem to have only a stabilizing role, whereas the C terminal disulfide bridge in both subunits evidently plays an important role in the early folding and activation of AGA. PMID- 8702914 TI - Identification of type I and type II serine/threonine kinase receptors for growth/differentiation factor-5. AB - Growth/differentiation factor-5 (GDF-5) is a member of the bone morphogenetic protein (BMP) family, which plays an important role in bone development in vivo. Mutations in the GDF-5 gene result in brachypodism in mice and Hunter-Thompson type chondrodysplasia in human. BMPs transduce their effects through binding to two different types of serine/threonine kinase receptors, type I and type II. However, binding abilities appear to be different among the members of the BMP family. BMP-4 binds to two different type I receptors, BMP receptors type IA (BMPR-IA) and type IB (BMPR-IB), and a type II receptor, BMP receptor type II (BMPR-II). In addition to these receptors, osteogenic protein-1 (OP-1, also known as BMP-7) binds to activin type I receptor (ActR-I) as well as activin type II receptors (ActR-II and ActR-IIB). Here we investigate the binding and signaling properties of GDF-5 through type I and type II receptors. GDF-5 induced alkaline phosphatase activity in a rat osteoprogenitor-like cell line, ROB-C26. 125I-GDF-5 bound to BMPR-IB and BMPR-II but not to BMPR-IA in ROB-C26 cells and other nontransfected cell lines. Analysis using COS-1 cells transfected with the receptor cDNAs revealed that GDF-5 bound to BMPR-IB but not to the other type I receptors when expressed alone. When COS-1 cells were transfected with type II receptor cDNAs, GDF-5 bound to ActR-II, ActR-IIB, and BMPR-II but not to transforming growth factor-beta type II receptor. In the presence of type II receptors, GDF-5 bound to different sets of type I receptors, but the binding was most efficient to BMPR-IB compared with the other type I receptors. Moreover, a transcriptional activation signal was efficiently transduced by BMPR-IB in the presence of BMPR-II or ActR-II after stimulation by GDF-5. These results suggest that BMPR-IB mediates certain signals for GDF-5 after forming the heteromeric complex with BMPR-II or ActR-II. PMID- 8702915 TI - Insulin signaling in mice expressing reduced levels of Syp. AB - Syp is a protein tyrosine phosphatase implicated in insulin and growth factor signaling. To evaluate the role of syp in insulin's regulation of plasma glucose, we generated knockout mice. Homozygous knockout mice die prior to day 10.5 of embryonic development. Hemizygous mice express half the levels of syp protein compared with their wild type littermates but do not display any gross morphological changes. Total body weight (age 2-10 weeks) and plasma insulin and glucose levels both in fasting and glucose-challenged states were comparable in the wild type and the hemizygous mice. No differences were observed in insulin induced glucose uptake in soleus muscle and epididymal fat; insulin inhibition of lipolysis was also similar. We injected insulin into the portal vein of the mice to examine upstream events of the insulin signaling cascade. Tyrosine phosphorylation of insulin receptor and insulin receptor substrate-1 (IRS-1) from hemizygous tissue was similar to that of wild type tissue. Association of the p85 subunit of phosphatidylinositol 3-kinase to IRS-1 increased an average of 2-fold in both groups. We did not observe an increase of IRS-1/syp association after insulin administration, but we did note a significant basal association in both wild type and hemizygous tissue. Our results do not support a major role for syp in the acute in vivo metabolic actions of insulin. PMID- 8702916 TI - Photoreceptor protein s26, a cone homologue of S-modulin in frog retina. AB - A frog retinal protein named s26 is a 26-kDa protein found during purification of S-modulin in frog retina (Kawamura, S. (1992) Photochem. Photobiol. 56, 1173 1180). To identify its role in frog retina, first s26 was purified to nearly homogeneity with three chromatographical steps. Based on the partial amino acid sequences of the proteolysed fragments of s26, we isolated cDNAs that encode s26. The analysis of its amino acid sequence revealed that s26 is an S-modulin-like protein, while it shows higher homology to visinin. Visinin is a Ca2+-binding protein reported to be present in chicken cones, but its localization in the retina had been a subject in dispute. The present study showed that s26 is present in cone photoreceptors. The study also showed that s26 inhibits phosphorylation of rhodopsin after a light flash at high Ca2+ concentrations as S modulin does. From these results, we concluded that s26 is a cone homologue of S modulin. The result is consistent with the idea that each type of photoreceptors expresses each cell-type specific version of phototransduction proteins. PMID- 8702917 TI - A potential SH3 domain-binding site in the Crk SH2 domain. AB - The Src homology 2 (SH2) domain of the mammalian adaptor protein Crk-II contains a proline-rich insert, predicted to lie within an extended DE loop, which is dispensable for phosphopeptide binding. Using the yeast two-hybrid system, this region of the Crk-II SH2 domain was found to interact with a subset of SH3 domains, notably the Abl SH3 domain. Furthermore, this proline-rich insert was found to modify the efficiency with which Crk-II was phosphorylated by the p140(c abl) tyrosine kinase. In vitro, the interaction of full-length non-phosphorylated Crk-II with a glutathione S-transferase-Abl SH3 domain fusion protein was very weak. However, phosphorylation of Crk-II on Tyr-221 which induces an intramolecular association with the SH2 domain, or addition of a phosphopeptide corresponding to the Crk-II Tyr-221 phosphorylation site, stimulated association of Crk-II with the Abl SH3 domain. NMR spectroscopic analysis showed that binding of the Tyr-221 phosphopeptide to the Crk SH2 domain induced a chemical shift change in Val-71, located in the proline-rich insert, indicative of a change in the structure of the proline-rich loop in response of Crk SH2-pTyr-221 interaction. These results suggest that the proline-rich insert in the Crk SH2 domain constitutes an SH3 domain-binding site that can be regulated by binding of a phosphopeptide ligand to the Crk SH2 domain. PMID- 8702918 TI - c-Jun is a downstream target for ceramide-activated protein phosphatase in A431 cells. AB - Stimulation of [3H]serine-labeled A431 cells with tumor necrosis factor-alpha (TNFalpha) or bacterial sphingomyelinase (SMase) resulted in a rapid decrease (approximately 50% by 15 min) in cellular [3H]sphingomyelin content and generation of the lipid moiety [3H]ceramide, which remained elevated 60 min later. Sphingomyelin hydrolysis in response to TNFalpha or bacterial SMase resulted in a time-dependent decrease in the phosphorylation state of c-Jun protein, an effect that was also observed in cells treated with the membrane permeable ceramide analogue N-hexanoylsphingosine (C6-ceramide). The rapid dephosphorylation of the c-Jun gene product in response to TNFalpha, SMase, or C6 ceramide was not observed in A431 cells treated with the serine-threonine phosphatase inhibitor okadaic acid. After the initial steps of previously described methods for the purification of a ceramide-activated protein phosphatase termed CAPP (Dobrowsky, R. T., Kamibayashi, C., Mumby, M. C., and Hannun, Y. A. (1993) J. Biol. Chem. 268, 15523-15530), we obtained a cytosolic fraction from A431 cells that specifically dephosphorylated 32Pi-labeled c-Jun protein used as substrate in an immunocomplex phosphatase assay. Phosphatase activity in vitro was apparent only in the presence of ceramide (5 micro) and was specifically abrogated when okadaic acid (1 n) was included in the immunocomplex phosphatase assay. These results provide strong evidence for c-Jun as a downstream target for CAPP activated in response to post-TNF signaling in A431 cells. PMID- 8702919 TI - Distinct tyrosine residues within the interleukin-2 receptor beta chain drive signal transduction specificity, redundancy, and diversity. AB - To explore the basis for interleukin (IL)-2 receptor (IL-2R) signaling specificity, the roles of tyrosine-based sequences located within the cytoplasmic tails of the beta and gammac chains were examined in the murine helper T cell line HT-2. Activation of the Janus kinase/signal transducer and activator of transcription (JAK/STAT) pathway, cellular proliferation, and the induction of various genes were monitored. All four of the cytoplasmic tyrosine residues as well as the distal portion of the gammac proved dispensable for the entire spectrum of IL-2R signaling responses studied. Conversely, select tyrosine residues within the beta chain were essential and differentially required for various signaling events. Specifically, activation of c-fos gene expression was found to occur exclusively through the most membrane proximal tyrosine, Tyr-338, whereas proliferation and the activation of STAT-5 were induced either through Tyr-338 or through the two C-terminal tyrosine residues, Tyr-392 and Tyr-510. These tyrosine residues mediated the induction of two different STAT-5 isoforms, which were found to form heterodimers upon receptor activation. In contrast to the tyrosine dependence of c-fos and STAT-5 induction, bcl-2 gene induction proceeded independently of all IL-2Rbeta tyrosine residues. Thus, the tyrosine based modules present within the IL-2Rbeta cytoplasmic tail play a critical role in IL-2R signaling, mediating specificity, redundancy, and multifunctionality. PMID- 8702920 TI - The extreme C terminus of primase is required for interaction with DnaB at the replication fork. AB - We have shown previously that a protein-protein interaction between DnaG and DnaB is required to attract the primase to the replication fork. This interaction was mediated by the C-terminal 16-kDa domain (p16) of the primase. A screen was developed that allowed the detection of mutant p16 proteins that did not interact with DnaB. Various mutagenesis protocols were used to localize this interaction domain to the extreme C terminus of the primase. A mutant primase missing only the C-terminal 16 amino acids was isolated and its activities examined. This mutant enzyme was fully active as a primase, but was incapable of interacting with DnaB. Thus, the mutant primase could not support DNA synthesis in either the general priming reaction or during phiX174 complementary strand DNA replication. Alanine cluster mutagenesis and deletion analysis in p16 allowed the further localization of the interaction domain to the extreme C-terminal 8 amino acids in primase. PMID- 8702921 TI - The interaction between helicase and primase sets the replication fork clock. AB - The synthesis of an Okazaki fragment occurs once every 1-2 s at the Escherichia coli replication fork and requires precise coordination of the enzymatic activities required. We have shown previously that the primase is recruited anew from solution for each cycle of Okazaki fragment synthesis and that association of primase with the replication fork is via a protein-protein interaction with the helicase, DnaB. We describe here mutant primases that have an altered interaction with DnaB and that direct the synthesis of Okazaki fragments of altered length compared to the wild-type. The mutant primases were deficient only in their ability to participate in replication reactions where their entry to the DNA was provided by the initial protein-protein interaction with DnaB. The primer synthesis capacity of these proteins remained unaffected, as was their ability to interact with the DNA polymerase III holoenzyme. Neither replication fork rate nor the efficiency of primer utilization was affected at replication forks programmed by the mutant enzymes. Thus, the interaction between DnaG and DnaB at the replication fork is the primary regulator of the cycle of Okazaki fragment synthesis. PMID- 8702922 TI - tau couples the leading- and lagging-strand polymerases at the Escherichia coli DNA replication fork. AB - Synthesis of an Okazaki fragment occurs once every 1 or 2 s at the Escherichia coli replication fork. To account for the rapid recycling required of the lagging strand polymerase, it has been proposed that it is held at the replication fork by protein-protein interactions with the leading-strand polymerase as part of a dimeric polymerase assembly. Solution studies showed that the replicative polymerase, the DNA polymerase III holoenzyme, was indeed a dimer with two catalytic cores held together by the tau subunit. However, the functionality of this arrangement at the replication fork has never been demonstrated. We showed previously that the lagging-strand polymerase acted processively during multiple rounds of Okazaki fragment synthesis, i.e. the same polymerase core assembly synthesized each and every fragment made by the fork. Using extreme dilution of active replication forks and the isolation of protein-DNA complexes capable of supporting coupled leading- and lagging-strand synthesis, we demonstrate here that this coupling of leading- and lagging-strand synthesis is, in fact, mediated by the tau subunit of the holoenzyme acting as a physical bridge between the core assemblies synthesizing the leading and lagging strands. PMID- 8702923 TI - An NF1-like protein functions as a repressor of the von Willebrand factor promoter. AB - The expression of the von Willebrand factor (vWf) gene is restricted to endothelial cells and megakaryocytes. We have previously reported the identification of a region of the vWf gene that regulates its cell-type-specific expression in cell culture. This region (spanning nucleotides -487 to +247) consists of a core promoter (spanning nucleotides -90 to +22), a positive regulatory region (spanning nucleotides +155 to +247), and a negative regulatory region spanning nucleotides -312 to -487. To identify the trans-acting factor(s) that interacts with the negative regulatory region, we carried out gel mobility and DNase1 footprint analyses of sequences -312 to -487. These analyses demonstrated that an NF1-like protein interacts with DNA sequences spanning -440 to -470 nucleotides in the negative regulatory region of the vWf promoter. Base substitution mutations of the NF1 binding site abolished the NF1-DNA interaction. Furthermore, mutation of the NF1 binding site in the promoter fragment (-487 to +155) that contained the core and the negative regulatory region resulted in activation of the mutant promoter in both endothelial and nonendothelial cells. The wild type promoter fragment (-487 to +155) was not activated in either cell type. These results demonstrate that an NF1-like protein functions as a repressor of vWf promoter activity. In contrast, the mutation of the same NF1 binding site, but now in the context of the larger 734-base pair endothelial cell-specific promoter fragment (-487 to +247), did not result in promoter activation in nonendothelial cells. The data indicate that there are additional repressor elements within the vWf promoter region suppressing its activity specifically, in nonendothelial cells, and suggest that there is a secondary repressor element(s) that is located in the terminal region of the first exon of this gene. PMID- 8702924 TI - Determination of the amino acid residue involved in [3H]beta-funaltrexamine covalent binding in the cloned rat mu-opioid receptor. AB - We previously demonstrated that [3H]beta-funaltrexamine ([3H]beta-FNA) labeled the rat mu opioid receptor expressed in Chinese hamster ovary cells with high specificity, and [3H]beta-FNA-labeled receptors migrated as one broad band with a mass of 80 kDa. In this study, we determined the region and then the amino acid residue of the mu receptor involved in the covalent binding of [3H]beta-FNA. [3H]beta-FNA-labeled receptors were solubilized and purified to approximately 10% purity by immunoaffinity chromatography with antibodies against a C-terminal domain peptide. The site of covalent bond formation was determined to be within Ala206-Met243 by CNBr cleavage of partially purified labeled mu receptors and determinations of sizes of labeled receptor fragments. The amino acid residue of beta-FNA covalent incorporation was then determined by site-directed mutagenesis studies within this region. Mutation of Lys233 to Ala, Arg, His, and Leu completely eliminated covalent binding of [3H]beta-FNA, although these mutants bound beta-FNA with high affinity. Mutations of other amino acid residues did not affect covalent binding of [3H]beta-FNA. These results indicate that [3H]beta-FNA binds covalently to Lys233. Since [3H]beta-FNA is a rigid molecule, the information will be very useful for molecular modeling of interaction between morphinans and the mu receptor. PMID- 8702925 TI - Modular structure of glucocorticoid receptor domains is not equivalent to functional independence. Stability and activity of the steroid binding domain are controlled by sequences in separate domains. AB - A long-standing conundrum of glucocorticoid receptors has been why the steroid binding domain is active in hybrid proteins but not in isolation. For this reason, the precise boundaries of the steroid binding domain have not been defined. These questions have now been systematically examined with a variety of receptor deletion constructs. Plasmids encoding amino acids 537-673 and 537-795 of the rat receptor did not yield stable proteins, while the fusion of receptor or non-receptor sequences upstream of 537-673 afforded stable proteins that did not bind steroid. Wild type steroid binding affinity could be obtained, however, when proteins such as beta-galactosidase or dihydrofolate reductase were fused upstream of receptor amino acids 537-795. Studies of a series of dhfr/receptor constructs with deletions at the amino- and carboxyl-terminal ends of the receptor sequence localized the boundaries of the steroid binding domain to 550 795. The absence of steroid binding upon deletion of sequences in the carboxyl terminal half of this domain was consistent with improperly folded receptor sequences. This conclusion was supported by analyses of the proteolysis and thermal stability of the mutant receptors. Thus, three independent regions appear to be required for the generation of the steroid binding form of receptors: 1) a protein sequence upstream of the steroid binding domain, which conveys stability to the steroid binding domain, 2) sequences of the carboxyl-terminal amino acids (674-795), which are required for the correct folding of the steroid binding domain, and 3) amino-terminal sequences (550-673), which may be sufficient for steroid binding after the entire steroid binding domain is properly folded. These results establish that the steroid binding domain of glucocorticoid receptors is not independently functional and illustrate the importance of both protein stability and protein folding when constructing mutant proteins. PMID- 8702926 TI - Analysis of M phase-specific phosphorylation of DNA topoisomerase II. AB - In mammalian cells, two isoforms of DNA topoisomerase II (topo II), topo IIalpha and topo IIbeta, are phosphorylated. The phosphorylation of topo IIbeta changes its apparent molecular mass determined by SDS-polyacrylamide gel electrophoresis from 180 to 190 kDa in mitotic cells, whereas topo IIalpha affects it only slightly (Kimura, K., Nozaki, N., Saijo, M., Kikuchi, A., Ui, M., and Enomoto, T. (1994) J. Biol. Chem. 269, 24523-24526). Here we examined the stability of the protein and the phosphate moiety of each topo II isoform, as the cells progressed from M to G1 phase. While its protein moiety remained intact, 75% of the phosphates attached to topo IIbeta were removed within 4 h after release from mitotic block. On the other hand, 35% of topo IIalpha protein and 52% of the attached phosphates disappeared. We verified that M phase-specific phosphorylation had no particular effect on the catalytic activities of both topo II isoforms after extensive phosphatase digestion. We also examined the binding of two isoforms to the nucleus or chromosomes. In logarithmically growing cells, both isoforms were extracted from nuclei at the same concentrations of NaCl. From the mitotic chromosomes, topo IIbeta was extracted at much lower concentrations of NaCl than topo IIalpha. PMID- 8702927 TI - 6B4 proteoglycan/phosphacan, an extracellular variant of receptor-like protein tyrosine phosphatase zeta/RPTPbeta, binds pleiotrophin/heparin-binding growth associated molecule (HB-GAM). AB - A major chondroitin sulfate proteoglycan in the brain, 6B4 proteoglycan/phosphacan, corresponds to the extracellular region of a receptor like protein-tyrosine phosphatase, PTPzeta/RPTPbeta. Here, we purified and characterized 6B4 proteoglycan-binding proteins from rat brain. From the CHAPS (3 [(3-cholamidopropyl)dimethylammonio]-1-propanesulfonic acid) extract of brain microsomal fractions, 18-, 28-, and 40-kDa proteins were specifically isolated using 6B4 proteoglycan-Sepharose. N-terminal amino acid sequencing identified the 18-kDa protein as pleiotrophin/heparin-binding growth-associated molecule (HB GAM). Scatchard analysis of 6B4 proteoglycan-pleiotrophin binding revealed low (Kd = 3 nM) and high (Kd = 0.25 nM) affinity binding sites. Chondroitinase ABC digestion of the proteoglycan decreased the binding affinities to a single value (Kd = 13 nM) without changing the number of binding sites. This suggested the presence of two subpopulations of the proteoglycan with different chondroitin sulfate structures. Heparin potently inhibited binding of 6B4 proteoglycan to pleiotrophin (IC50 = 3.5 ng/ml). Heparan sulfate and chondroitin sulfate C inhibited moderately (IC50 = 150 and 400 ng/ml, respectively), but, in contrast, chondroitin sulfate A and keratan sulfate were poor inhibitors (IC50 > 100 microg/ml). Immunofluorescence and immunoblotting analyses indicated that both 6B4 proteoglycan and PTPzeta are located on cortical neurons. Anti-6B4 proteoglycan antibody added to the culture medium suppressed pleiotrophin-induced neurite outgrowth of cortical neurons. These results suggested that interaction between 6B4 proteoglycan and pleiotrophin is required for the action of pleiotrophin, and chondroitin sulfate chains on 6B4 proteoglycan play regulatory roles in its binding. PMID- 8702928 TI - RNA polymerase signals UvrAB landing sites. AB - Transcription when coupled to nucleotide excision repair specifies the location in active genes where preferential DNA repair is to take place. During DNA damage induced recruitment of RNA polymerase (RNAP), there is a physical association of the beta subunit of Escherichia coli RNAP and the UvrA component of the repair apparatus (G. C. Lin and L. Grossman, submitted for publication). This molecular affinity is reflected in the ability of the RNAP to increase, in a promoter dependent manner, DNA supercoiling by the UvrAB complex. In the presence of the RNAP, the UvrAB complex is able to bind to promoter regions and to translocate in a 5' to 3' direction along the non-transcribed strand. As a consequence of this helicase-catalyzed translocation, preferential incision of DNA damaged sites occurs downstream on the transcribed strand. Because of the helicase directionality, the initial binding of the UvrAB complex to the transcribed strand would inevitably lead to its collision with the RNAP. These results imply that the RNAP-induced DNA structure in the vicinity of the transcription start site signals a landing or entry site for the UvrAB complex on DNA. PMID- 8702929 TI - The binding of UvrAB proteins to bubble and loop regions in duplex DNA. AB - Based on the binding of the UvrAB complex to a promoter region in transcription open complexes (Ahn, B., and Grossman, L. (1996) J. Biol. Chem. 271, 21453-21461) and the requirement of a single-stranded region for UvrAB helicase activity, we examined the binding of UvrAB proteins to synthetic bubble or loop regions in duplex DNA and the role of these regions in translocation of the UvrAB complex as well as incision of DNA damage. We found that the UvrAB complex was able to bind to bubble and loop regions with an affinity similar to that for damaged DNA in the absence of RNAP. The preferential recognition and incision of damaged sites by the UvrAB complex was observed downstream of the bubble or loop region in the strand complementary to the strand along which the UvrAB complex translocates. These results imply that the bubble region generated in duplex DNA by RNAP serves as a preferred entry site for the translocation of the UvrAB complex, and that preferential binding and unidirectional translocation of the UvrAB complex predetermine where incision is to occur. PMID- 8702930 TI - Reduced requirement of mitogen-activated protein kinase (MAPK) activity for entry into the S phase of the cell cycle in Swiss 3T3 fibroblasts stimulated by bombesin and insulin. AB - Bombesin induced a marked and persistent activation of the mitogen-activated protein kinase kinase-1 (MEK-1), p42(mapk) and p90(rsk) in Swiss 3T3 cells by a pathway that was independent of p74(raf-1) but dependent on the activity of protein kinase C. Pretreatment of the cells with a specific inhibitor of MEK-1, PD 098059, markedly reduced the early and abolished the sustained phase of bombesin-induced p42(mapk) activation. In addition, PD 098059 prevented bombesin induced DNA synthesis and progression of the cells through the cell cycle, indicating that the mitogenic effect of bombesin is dependent on the activation of p42(mapk). However, in the presence of insulin, which neither stimulated p42(mapk) activation nor DNA synthesis on its own in Swiss 3T3 cells, bombesin potently stimulated DNA synthesis even at concentrations of PD 098059 (15 microM) that completely abolished the mitogenic effect of bombesin alone. Furthermore, Swiss 3T3 cells stably transfected with interfering mutants of MEK-1 showed a marked decrease in the mitogenic effect of bombesin. In contrast, the combination of bombesin and insulin strongly stimulated DNA synthesis in these cells to levels comparable with that obtained in the wild type cells. Thus, our data demonstrate that insulin dramatically reduced the requirement for the mitogen activated protein kinase pathway for reinitiation of DNA synthesis in bombesin treated Swiss 3T3 cells and consequently indicate that the contribution of the mitogen-activated protein kinase cascade to mitogenesis depends on the combination of extracellular signals that are used to stimulate these cells. PMID- 8702931 TI - Protein kinase C deficiency blocks recovery from agonist-induced desensitization. AB - Protein phosphorylation is central to agonist-induced attenuation of the function of G-protein-linked receptors. Stable expression of RNA antisense to specific protein kinase mRNAs permitted analysis of loss-of-function mutants of A431 human epidermoid carcinoma cells, lacking protein kinase A, protein kinase C, or beta adrenergic receptor kinase. Deficiency of protein kinase C, but not the others, amplified rather than attenuated agonist-induced desensitization. In wild-type cells, the t1/2 for recovery from desensitization was approximately 25 min following removal of agonist. In the protein kinase C-deficient cells, no resensitization was observed even 60 min after agonist removal. Like protein kinase C-deficiency, inhibition of protein kinase C with bisindolylmaleimide or calphostin C blocked resensitization. Resensitization was suppressed by FK506, an inhibitor of protein phosphatase 2B, mimicking protein kinase C-deficiency, but in a non-additive manner. The data reveal protein kinase C and protein phosphatase 2B to be critical elements of resensitization. PMID- 8702932 TI - CCAAT/enhancer-binding protein alpha activation of the rat growth hormone promoter in pituitary progenitor GHFT1-5 cells. AB - High level, anterior pituitary-specific expression of the rat growth hormone (rGH) promoter requires cooperative actions of several different transcription factors. Previously, we described a series of multisubunit, tissue-general, transcription factor complexes that bound to the GHF3 activation site and strongly regulated rGH promoter activity. A 43-kDa DNA-binding subunit common to each of the different GHF3 complexes is identified here as the transcription factor, CCAAT/Enhancer-binding Protein alpha (C/EBPalpha). In human monocyte U937 cells, which do not express the endogenous or transfected GH genes, co-expression of C/EBPalpha and Pit-1 synergistically activated the transfected rGH promoter. Full-length C/EBPalpha was present in the GH-secreting GC, and prolactin secreting 235-1, pituitary cell lines, but not in GHFT1-5 cells, which are transformed at a stage in development immediately prior to GH expression. Transient expression of C/EBPalpha in GHFT1-5 cells strongly activated the co transfected rGH promoter through the GHF3 binding site; a second activation site mapped to evolutionary conserved GH promoter sequences between -106 and -33. C/EBPalpha activation was synergistic with phorbol 12-myristate 13-acetate and forskolin, activators of protein kinases C and A, respectively. Thus, C/EBPalpha is an important regulator of rGH promoter activity that appears to function in synergy with Pit-1, activators of A and C protein kinases and possibly other factors. PMID- 8702933 TI - Palmitoylated cysteine 341 modulates phosphorylation of the beta2-adrenergic receptor by the cAMP-dependent protein kinase. AB - We previously showed that substitution of a glycine residue for the palmitoylated cysteine 341 of the human beta2-adrenergic receptor (Gly341beta2AR), increases the basal level of the receptor phosphorylation and reduces its ability to functionally interact with Gs. In the present study, we show that additional mutation of serines 345 and 346 (Ala345,346Gly341beta2AR) restored normal phosphorylation and receptor-Gs coupling, thus suggesting that the increased phosphorylation of this site, rather than the lack of palmitoylation per se, is responsible for the poor coupling of the unpalmitoylated receptor. This is supported by the observation that chemical depalmitoylation of purified beta2AR did not affect the ability of the receptor to stimulate adenylyl cyclase in reconstitution assays. Furthermore, mutation of Ser345,346 in a wild type receptor background (Ala345,346beta2AR) significantly decreased the rate of agonist-promoted desensitization of the receptor-stimulated adenylyl cyclase activity, supporting a role for this phosphorylation site in regulating the functional coupling of the receptor. Since serines 345 and 346 are located in a putative cyclic AMP-dependent protein kinase (PKA) phosphorylation site immediately downstream of the palmitoylated cysteine 341, the hypothesis that the accessibility of this site may be regulated by the receptor palmitoylation state was further assessed in vitro. In membrane phosphorylation assays, Gly341beta2AR was found to be a better substrate for PKA than the wild type receptor, thus supporting the notion that palmitoylation restrains access of the phosphorylation site to the enzyme. Taken together, the data demonstrate that palmitoylation of cysteine 341 controls the phosphorylation state of the PKA site located in the carboxyl tail of the beta2AR and by doing so modulates the responsiveness of the receptor. PMID- 8702934 TI - Cleavage arrest of early frog embryos by the G protein-activated protein kinase PAK I. AB - PAK I is a member of the PAK (p21-activated protein kinase) family and is activated by Cdc42 (Jakobi, R., Chen, C.-J., Tuazon, P. T., and Traugh, J. A. (1996) J. Biol. Chem. 271, 6206-6211). To examine the effects of PAK I on cleavage arrest, subfemtomole amounts of endogenously active (58 kDa) and inactive (60 kDa) PAK I and a tryptic peptide (37 kDa) containing the active catalytic domain were injected into one blastomere of 2-cell frog embryos. Active PAK I resulted in cleavage arrest in the injected blastomere at mitotic metaphase, whereas the uninjected blastomere progressed through mid- to late cleavage. Injection of other protein kinases at similar concentrations had no effect on cleavage. Endogenous PAK I was highly active in frog oocytes, and antibody to PAK I reacted specifically with protein of 58-60 kDa. PAK I protein was decreased at 60 min post-fertilization, with little or no PAK I protein or activity detectable at 80 min post-fertilization or in 2-cell embryos. At the 4 cell stage PAK I protein increased, but the protein kinase was present primarily as an inactive form. Rac2 and Cdc42, but not Rac 1, were identified in oocytes and throughout early embryo development. Thus, PAK I appears to be a potent cytostatic protein kinase involved in maintaining cells in a non-dividing state. PAK I activity is high in oocytes and appears to be regulated by degradation/synthesis and through autophosphorylation via binding of Cdc42. PAK I may act through regulation of the stress-activated protein kinase signaling pathway and/or by direct regulation of multiple metabolic pathways. PMID- 8702935 TI - Cytosolic phospholipase A2 (PLA2), but not secretory PLA2, potentiates hydrogen peroxide cytotoxicity in kidney epithelial cells. AB - Phospholipase A2 (PLA2) and reactive oxygen species have been implicated both individually and synergistically in various forms of cellular injury. The form(s) of PLA2 important for cell injury and the implications of enhanced activity of the enzyme, however, have not been discerned. Previous studies reveal an increase in PLA2 activity associated with cell injury, but this association does not establish a causal relationship between the increase in activity and the injury. LLC-PK1 cell lines were created that express either the cytosolic PLA2 or a group II PLA2. The susceptibility of these cells to hydrogen peroxide toxicity was determined in order to evaluate the relative importance of these two forms of PLA2 in oxidant injury. Expression of cytosolic PLA2 in the LLC-cPLA2 cell line was associated with a 50-fold increase in PLA2 activity in the cytosolic fraction, an increase in agonist-stimulated arachidonate release, and immunodetection of the cytosolic PLA2 protein that was undetectable in control cells. Exposure to hydrogen peroxide or menadione, but not mercuric chloride, resulted in significantly greater lactate dehydrogenase release in LLC-cPLA2 cells when compared with control cells. Exogenous arachidonic acid (150 microM) did not enhance hydrogen peroxide-induced injury. The intracellular calcium chelator, 1,2-bis-(o-aminophenoxy)ethane-N,N,N', N'-tetraacetic acid/tetra(acetoxymethyl) ester, protected the cells against injury, but the calcium ionophore, A23187, did not increase injury. Glycine conferred no protective effect against hydrogen peroxide toxicity. By contrast to these results with cytosolic PLA2-expressing cells, secretory PLA2 expression to very high levels did not increase susceptibility to hydrogen peroxide. Thus, cytosolic PLA2 may an be an important mediator of oxidant damage to renal epithelial cells. PMID- 8702936 TI - Molecular cloning of a novel T cell-directed CC chemokine expressed in thymus by signal sequence trap using Epstein-Barr virus vector. AB - Precursors of most secreted and cell surface molecules carry signal sequences at their amino termini. Here we describe an efficient signal sequence trap method and isolation of a novel CC chemokine. An expression library was constructed by inserting 5' portion-enriched cDNAs from phytohemagglutinin-stimulated peripheral blood mononuclear cells into upstream of signal sequence-deleted CD4 cDNA in an Epstein-Barr virus shuttle vector. After electroporation into Raji cells, CD4 antigen-positive cells were enriched by repeated cell sorting and plasmids were recovered in Escherichia coli. Out of 100 plasmid clones examined, 42 clones directed expression of CD4 antigen on the cell surface. Among them were signal sequences of CD6, beta2-microglobulin, MGC-24, and T cell receptor epsilon-chain, and at least four novel potential signal sequences. A cDNA clone encoding a novel CC chemokine was isolated by using one of the trapped fragments. The gene designated as TARC from Thymus and Activation-Regulated Chemokine was expressed transiently in phytohemagglutinin-stimulated peripheral blood mononuclear cells and constitutively in thymus. Radiolabeled recombinant TARC specifically bound to T cell lines and peripheral T cells but not to monocytes or granulocytes. The binding of radiolabeled TARC to the high-affinity receptor (Kd, 2.1 nM) on Jurkat was displaced by TARC but not by interleukin-8, MIP-1alpha, RANTES, or MCP-1. TARC also bound to the promiscuous chemokine receptor on erythrocytes (Kd, 17 nM). TARC induced chemotaxis in T cell lines Hut78 and Hut102. Pretreatment of Hut78 with pertussis toxin abolished the TARC-induced cell migration. Collectively, T cells express a highly selective receptor for TARC that is coupled to pertussis toxin-sensitive G-protein. TARC may a factor playing important roles in T cell development in thymus as well as in trafficking and activation of mature T cells. PMID- 8702937 TI - Differential effects of protein kinase C activation on calcium storage and capacitative calcium entry in NIH 3T3 cells. AB - In NIH 3T3 cells, treatment with phorbol 12-myristate 13-acetate (PMA) reduced the release of Ca2+ by thapsigargin, but did not activate Ca2+ entry; Ca2+ influx was triggered after the residual pool was emptied by thapsigargin, and this Ca2+ influx was similar to that induced by thapsigargin in control cells. The effect of PMA was due to decreased Ca2+ storage because 1) Ca2+ release by ionomycin was similarly affected by PMA, and in both control and PMA-treated cells, ionomycin did not release Ca2+ following thapsigargin treatment; 2) PMA reduced 45Ca2+ accumulation; and 3) studies with Ca2+ indicator compartmentalized into the endoplasmic reticulum indicated that stored Ca2+ was reduced by PMA. Although PMA did not itself activate Ca2+ entry, PMA potentiated Ca2+ entry with low concentrations of cyclopiazonic acid. With a somewhat higher concentration of cyclopiazonic acid, PMA had no effect on calcium entry. Thus, protein kinase C has two apparent actions on calcium signaling in NIH 3T3 cells: 1) reduced intracellular Ca2+ storage capacity and 2) augmented calcium entry with submaximal intracellular Ca2+ pool depletion. These actions indicate a complex and potentially important role for the protein kinase C system in calcium homeostasis in this cell type. PMID- 8702938 TI - Two distinct modes of strand unlinking during theta-type DNA replication. AB - DNA gyrase and topoisomerase IV (Topo IV) are type II bacterial DNA topoisomerases that show a high degree of similarity to each other yet appear to have different functions during DNA replication. We show here that the torsional stress generated during theta-type DNA replication in vitro can take the form of either positive supercoils ahead of the replication fork or catenane-like right handed windings (precatenanes) of the two partially replicated duplexes behind the fork. Gyrase prefers to act on the former, whereas Topo IV prefers the latter. Removal of either form of positive winding can support nascent chain elongation, but only precatenane removal can support the final stages of DNA replication: processing of the late intermediate and daughter chromosome decatenation. PMID- 8702939 TI - The cloned thrombin receptor is necessary and sufficient for activation of mitogen-activated protein kinase and mitogenesis in mouse lung fibroblasts. Loss of responses in fibroblasts from receptor knockout mice. AB - The mitogenic activity of thrombin on fibroblasts and smooth muscle cells may contribute to embryonic development and normal wound healing, and it may also play a role in pathological responses to vascular injury. To examine the importance of thrombin signaling in vivo and to define the cloned thrombin receptor's role, we disrupted the thrombin receptor gene (tr) in mice. Platelets from tr-/- mice responded normally to thrombin, but tr-/- fibroblasts showed no thrombin-induced calcium mobilization or phosphoinositide hydrolysis. Thus distinct thrombin receptors act in different tissues. This study focuses on the role of the thrombin receptor in thrombin-induced mitogenesis and mitogen activated protein (MAP) kinase activation in mesenchymal cells. Thrombin and thrombin receptor agonist peptide both stimulated DNA synthesis and MAP kinase activation in fibroblasts derived from wild-type mice. These responses were selectively lost in fibroblasts from tr-/- mice. Activation of the cloned thrombin receptor is therefore necessary and sufficient for thrombin-induced mitogenesis and MAP kinase activation in mouse lung fibroblasts. The tr-/- mouse thus provides a valuable model for defining the role of thrombin-induced proliferative events in vivo. Because thrombin-induced MAP kinase activation was attributable to a single receptor expressed at natural levels, mouse lung fibroblasts presented an opportunity to define the pathways that normally mediate activation of MAP kinase by the thrombin receptor. Elimination of phorbol sensitive protein kinase C by prolonged exposure to phorbol ester only partially inhibited MAP kinase activation by thrombin but completely blocked c-Raf kinase activation. Pertussis toxin partially inhibited MAP kinase activation by thrombin but had no significant effect on c-Raf kinase activation. Thus in mouse lung fibroblasts, one thrombin receptor utilizes two pathways for MAP kinase activation: one is protein kinase C- and c-Raf dependent, and a second is Gi dependent and c-Raf-independent. PMID- 8702940 TI - A unique phosphorylation-dependent mechanism for the activation of Ca2+/calmodulin-dependent protein kinase type IV/GR. AB - The activity of the Ca2+/calmodulin-dependent protein kinase IV/Gr (CaMKIV/Gr) is shown to be strictly regulated by phosphorylation of three residues both in vitro and in response to antigen receptor-mediated signaling in lymphocytes. One residue, Thr-200, is indispensable for enhancement of Ca2+/calmodulin-dependent basal activity by CaMKIV/Gr kinase. This event requires Ca2+/calmodulin in the full-length CaMKIV/Gr but is Ca2+/calmodulin-independent when a truncated version of CaMKIV/Gr is used as a substrate (DeltaCaMKIV/Gr1-317 (Delta1-317)). The other two residues, Ser12 and Ser13, are apparently autophosphorylated by the Ca2+/calmodulin-bound CaMKIV/Gr. Phosphorylation of neither Ser12-Ser13 nor Thr312 (the residue in a homologous position to Thr286 of CaMKIIalpha influences the development of Ca2+/calmodulin-independent activity or any other property of CaMKIV/Gr examined. Similarly, removal of the NH2-terminal 20 amino acids has no effect on the activation or function of CaMKIV/Gr. However, mutation of both Ser12 and Ser13 residues to Ala in Delta1-317 completely abrogates activity, while individual substitutions have no effect. These results indicate that the NH2-terminal Ser cluster mediates a novel type of intrasteric inhibition and suggest that three events are required for CaMKIV/Gr activation: 1) Ca2+/calmodulin binding; 2) phosphorylation of the Ca2+/calmodulin-bound enzyme on Thr200 by a Ca2+/calmodulin-dependent protein kinase kinase; and 3) autophosphorylation of Ser12-Ser13. This three-step requirement is unique among the multifunctional Ca2+/calmodulin-dependent kinases. PMID- 8702941 TI - Amanitin greatly reduces the rate of transcription by RNA polymerase II ternary complexes but fails to inhibit some transcript cleavage modes. AB - The toxin alpha-amanitin is frequently employed to completely block RNA synthesis by RNA polymerase II. However, we find that polymerase II ternary transcription complexes stalled by the absence of NTPs resume RNA synthesis when NTPs and amanitin are added. Chain elongation with amanitin can continue for hours at approximately 1% of the normal rate. Amanitin also greatly slows pyrophosphorolysis by elongation-competent complexes. Complexes which are arrested (that is, which have paused in transcription for long periods in the presence of excess NTPs) are essentially incapable of resuming transcription in the presence of alpha-amanitin. Complexes traversing sequences that can provoke arrest are much more likely to stop transcription in the presence of the toxin. The substitution of IMP for GMP at the 3' end of the nascent RNA greatly increases the sensitivity of stalled transcription complexes to amanitin. Neither arrested nor stalled complexes display detectable SII-mediated transcript cleavage following amanitin treatment. However, arrested complexes possess a low level, intrinsic transcript cleavage activity which is completely amanitin resistant; furthermore, pyrophosphorolytic transcript cleavage in arrested complexes is not affected by amanitin. PMID- 8702942 TI - Conformations of the nucleotide and polypeptide binding domains of a cytosolic Hsp70 molecular chaperone are coupled. AB - 70-kDa heat shock protein (Hsp70) molecular chaperones are ATPases that participate in protein folding by regulating protein-protein interactions. ATP binds to the highly conserved amino-terminal domain, whereas polypeptides bind to the less conserved carboxyl-terminal domain. These domains are functionally coupled. Polypeptides were previously shown to dissociate from Hsp70s upon ATP binding and to stimulate ATPase activity. We probed the structure of the yeast cytosolic Hsp70 Ssa1p using limited proteolysis to determine whether the conformations of its nucleotide and polypeptide binding domains are also coupled. Ssa1p adopted three distinct conformations, nucleotide-free, ADP-dependent, and ATP-dependent. Complete conformational changes required K+ and Mg2+. Using amino terminal sequencing, ATP-agarose chromatography, and a carboxyl-terminal-specific antibody, we mapped the locations of the major proteolytic fragments. Nucleotides altered the conformations of both the nucleotide and polypeptide binding domains. Similarly, a polypeptide altered the conformations of both domains. These results indicate that the conformations of the nucleotide and polypeptide binding domains are coupled. PMID- 8702943 TI - Triple helix formation of procollagen type I can occur at the rough endoplasmic reticulum membrane. AB - One key problem in understanding the biosynthesis of collagens remains the assembly of the three alpha-chains. How and when are the different gene products selected, aligned, and folded into a triple helix? As the spatial arrangement during biosynthesis might be important, we concentrated on whether the rough endoplasmic reticular membrane is involved in this process. Microsomes were prepared from biosynthetically labeled chick tendon fibroblasts. Vesicles were spread as a monomolecular film which was then transferred over several compartments of a filmbalance containing fresh subphase. Fluorograms of the surface film showed that the monolayer contains procollagen chains. When the monolayer was transferred onto a chymotrypsin/trypsin-containing subphase, the gel bands of the proalpha-chains were shifted into the position of mature alpha chains, indicating that only the propeptides were digested and the collagenous regions were protected due to triple helix formation. Our results suggest that newly synthesized proalpha-chains can associate as trimers and fold into a triple helical conformation while they are still associated with the membranes of the rough endoplasmic reticulum. These processes also occur when interchain disulfide linkage is inhibited, indicating that chain selection and registration is not dependent on formation of covalent bonds among the carboxyl propeptides. PMID- 8702944 TI - Prolactin inhibits epidermal growth factor-induced Ras-MAPK signaling in mammary epithelial cells. AB - Previously, our laboratory has shown that prolactin (PRL) inhibits epidermal growth factor (EGF)-induced DNA synthesis. One pathway for the initiation of DNA synthesis is EGF-receptor (EGF-R) signaling through Ras and mitogen-activated protein kinase (MAPK). To determine the effects of PRL on EGF-induced MAPK activation and phosphorylation, MAPK or phosphotyrosine (Tyr(P)) was immunoprecipitated from normal murine mammary epithelial (NMuMG) cells treated with PRL (100 ng/ml) and/or EGF (10 ng/ml) for 10-min periods. EGF-induced phosphorylation and activation were then examined by Western analysis and a myelin basic protein (MBP)-specific kinase assay. The p42 isoform of MAPK showed a distinct decrease in activity and phosphorylation when cells were treated with PRL. Concluding that PRL affects EGF signaling upstream of MAPK, we examined the effect of PRL on EGF-induced Ras activity. NMuMG cells were incubated with [32P]orthophosphoric acid, treated as described above, immunoprecipitated with an antibody specific to Ras, and nucleotides were eluted and separated by TLC. Ras activity as measured by GTP:GDP ratio was increased by EGF, but not by PRL. Additionally, PRL in combination with EGF abolished the ability of EGF to induce Ras activity. Those studies suggest that PRL alters the EGF signaling pathway upstream of Ras. Because Ras activation by EGF involves EGF-stimulated association of EGF-R with Grb2, the EGF-R was immunoprecipitated and a Western blot was probed for Grb2. As expected we found that EGF stimulated an association of EGF-R with Grb2, PRL, however, blocked this association. When we looked at the ability of Shc to associate with the EGF-R, we found that PRL and EGF had little effect on this association. The studies demonstrate that PRL either directly or indirectly inhibits the ability of EGF to induce EGF-R association with Grb2, to activate Ras, and to activate and phosphorylate MAPK. PMID- 8702945 TI - The amino-terminal charge and core region hydrophobicity interdependently contribute to the function of signal sequences. AB - We have constructed a series of signal sequence mutants that contain negatively charged amino termini and simplified core regions of varying hydrophobicity levels. This series provides a means of exploring the relative roles of the amino terminus and the hydrophobic core region during transport. The signal peptides with highly hydrophobic core regions support a rapid rate of transport in the presence of a negatively charged amino terminus. We have found that these negatively charged mutants are secreted in a manner similar to the wild-type signal sequence; sodium azide and carbonyl cyanide 3-chlorophenylhydrazone treatments indicate that the negatively charged mutants depend on SecA and the protonmotive force, respectively. These same mutants also demonstrate reduced competition with coexpressed beta-lactamase, reflecting the lower overall affinity for the transport pathway due to the net negative charge at the amino terminus. In addition, the pronounced effects of introducing three negative charges support the conclusion that the two regions function in a concerted manner. PMID- 8702946 TI - Purification and structural characterization of a filamentous, mucin-like proteophosphoglycan secreted by Leishmania parasites. AB - Parasitic protozoa of the genus Leishmania secrete a filamentous macromolecule that forms networks and appears to be associated with cell aggregation. We report here the purification of this parasite antigen from Leishmania major culture supernatant and its compositional (75.6% carbohydrate, 20% phosphate, 4.4% amino acids, w/w), structural, and ultrastructural characterization as a highly unusual proteophosphoglycan (PPG). Mild acid hydrolysis, which cleaves preferentially hexose 1-phosphate bonds, releases the PPG glycans. Their structures are Galbeta1 4Man, Manalpha1-2Man, Galbeta1-3Galbeta1-4Man, PO4-6(Galbeta1-3)0-2Galbeta1-4Man, and PO4-6(Arabeta1-2Galbeta1-3)Galbeta1-4Man. These glycans are also components of the parasite glycolipid lipophosphoglycan, but their relative abundance and structural organization in PPG are different. Some of them represent novel forms of protein glycosylation. 31P NMR on native PPG demonstrates that phosphate is exclusively in phosphodiester bonds and that the basic structure R-Manalpha1-PO4 6-Gal-R connects the glycans. A phosphodiester linkage to phosphoserine (most likely R-Manalpha1-PO4-Ser) anchors the PPG oligosaccharides to the polypeptide. PPG has a unique amino acid composition; glycosylated phosphoserine (>43 mol %), serine, alanine, and proline account for more than 87 mol % and appear to be clustered in large proteinase-resistant domains. Electron microscopy of purified PPG reveals cable-like, flexible, long (to 6 microm), and unbranched filaments. The overall structure of PPG shows many similarities to mammalian mucins. Potential functions of this novel mucin-like molecule for the parasites are discussed. PMID- 8702947 TI - Rho-dependent termination of transcription is governed primarily by the upstream Rho utilization (rut) sequences of a terminator. AB - A Rho-dependent transcription terminator in Escherichia coli DNA consists of an upstream part for Rho utilization (rut) and the transcription stop point (tsp) region. To test the role of the tsp region variants of the coliphage lambda cro gene terminator, tR1, containing inserts of non-terminator sequences between its rut and tsp regions were tested for termination function. The results showed that termination occurred with high efficiency at multiple sites in each of the new sequences with the positions of the sites coinciding with transcriptional pause points in the insert sequence and that the efficiency of termination was not directly proportional to the extent of pausing at those points. Thus, in contrast to the rut sequences, which are relatively rare in DNA, many different sequence segments can function as a tsp region. Studies with isolated transcripts showed that a rut element and sequences 3' of the rut element were both needed to activate ATP hydrolysis by Rho factor with the degree of activation depending on the length and the specific sequence of the 3' segment. These results support models for Rho action in which ATP hydrolysis is coupled to interactions of Rho protein with RNA 3' of the rut region. PMID- 8702948 TI - Intracellular trafficking of cholesterol monitored with a cyclodextrin. AB - The sterol binding agent 2-hydroxypropyl-beta-cyclodextrin is shown to be a convenient and useful experimental tool to probe intracellular pathways of cholesterol transport. Biochemical and cytochemical studies reveal that cyclodextrin specifically removes plasma membrane cholesterol. Depletion of plasma membrane sphingomyelin greatly accelerated cyclodextrin-mediated cholesterol removal. Cholesterol arriving at the plasma membrane from lysosomes and the endoplasmic reticulum was also removed by cyclodextrin. Cellular cholesterol esterification linked to the mobilization of cholesterol from lysosomes was strongly attenuated by cyclodextrin, suggesting that the major portion of endocytosed cholesterol is delivered from lysosomes to the endoplasmic reticulum via the plasma membrane. Evidence for translocation of lysosomal cholesterol to the endoplasmic reticulum by a plasma membrane-independent pathway is provided by the finding that cyclodextrin loses its ability to suppress esterification when plasma membrane sphingomyelin is depleted. The Golgi apparatus appears to play an active role in directing the relocation of lysosomal cholesterol to the plasma membrane since brefeldin A also abrogated cyclodextrin mediated suppression of cholesterol esterification. Using cyclodextrin we further show that attenuated esterification of lysosomal cholesterol in Niemann-Pick C cells reflects defective translocation of cholesterol to the plasma membrane that may be linked to abnormal Golgi trafficking. PMID- 8702949 TI - Binding to the platelet-derived growth factor receptor transiently activates the p85alpha-p110alpha phosphoinositide 3-kinase complex in vivo. AB - Ligand stimulation of the platelet-derived growth factor (PDGF) receptor results in its association with phosphoinositide 3-kinase activity and a corresponding synthesis of 3'-phosphorylated lipids. Early studies that examined this interaction in vivo employed anti-phosphotyrosine antiserum or antiserum against the PDGF receptor. The recent identification of multiple isoforms of both the regulatory and the catalytic subunit of the enzyme have led us to utilize antisera against p85alpha and p110alpha to characterize the association of this particular phosphoinositide 3-kinase complex with the PDGF receptor following ligand stimulation of murine fibroblasts. Both the p85alpha and p110alpha subunits rapidly associated with the ligand-activated receptor resulting in a transient, 2-fold increase in the total pool of p110alpha lipid kinase activity. This association was stable for 15 min after initial stimulation. Subsequently, both subunits began to dissociate from the receptor with similar kinetics. By 60 min this process was complete, demonstrating that p85alpha and p110alpha both associate with the receptor and dissociate from the receptor as a dimeric complex. At this time, marked PDGF receptor down-regulation was observed. Immunoprecipitation from metabolically labeled cells revealed that p85alpha is constitutively phosphorylated on serine residues in quiescent cultures. Upon PDGF stimulation, this phosphorylation upon serine residues was maintained in addition to tyrosine phosphorylation of this subunit. No phosphorylation of the p110alpha subunit was detected in either quiescent or PDGF-stimulated cells. Quantitation of Western blot analysis demonstrated that only 5% of the total pool of p85alpha associated with the PDGF receptor upon ligand stimulation. The 2-fold increase in the lipid kinase activity measured in immunoprecipitates using either anti p85alpha or anti-p110alpha antiserum therefore reflects a far greater increase in the specific activity of the enzyme upon its association with the PDGF receptor. PMID- 8702950 TI - Interaction of the N-methyl-D-aspartate receptor complex with a novel synapse associated protein, SAP102. AB - Ionotropic glutamate receptors are known to cluster at high concentration on the postsynaptic membrane of excitatory synapses, but the mechanism by which this occurs is poorly understood. Studies on the neuromuscular junction and central inhibitory synapses suggest that clustering of neurotransmitter receptors requires its interaction with a cytoplasmic protein. Recently, in vitro studies have shown that members of the N-methyl--aspartate (NMDA) class of glutamate receptors interact with a synapse-associated protein, SAP90 (PSD-95). However, evidence for the in vivo interaction of NMDA receptors with SAPs is still lacking. In the present study, we demonstrate the specific interaction between SAP102, a novel synapse-associated protein, and the NMDA receptor complex from the rat cortical synaptic plasma membranes using co-immunoprecipitation techniques. No association was observed between SAP102 and GluR1, a member of the alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate class of glutamate receptors. To identify the domain on the NMDA receptor responsible for this interaction, we constructed hexahistidine fusion proteins from different regions of the NR1a and NR2 subunits of the NMDA receptor. Immunoblot overlay experiments showed that while the C-terminal domain of the NR2 subunit displayed strong binding, the NR1a intracellular C-terminal tail did not interact with SAP102. The site of interaction was more precisely located to the last 20 amino acids of the NR2 subunit as indicated by the interaction of the synthetic peptide with SAP102. In summary, we demonstrate here for the first time an in vivo interaction between the native NMDA receptor complex and a synapse-associated protein. These results suggest that SAP102 may play an important role in NMDA receptor clustering and immobilization at excitatory synapses. PMID- 8702951 TI - Loss of function of cytochrome c in Jurkat cells undergoing fas-mediated apoptosis. AB - Mitochondrial function was examined in Jurkat cells undergoing Fas-mediated apoptosis. With succinate or ascorbate/tetramethylphenylenediamine as substrate, oxygen uptake by digitonin-permeabilized apoptotic mitochondria was greatly decreased as compared with control. Assessment of the function of the cytochrome c-cytochrome oxidase segment of the electron transport chain of apoptotic mitochondria showed that the activity of cytochrome oxidase appeared to be normal, but that of cytochrome c was greatly diminished. A death protease was found to participate in the events leading to the loss of cytochrome c activity, but the cytochrome did not seem to be extensively degraded during the course of apoptosis. Our results suggest that a rapid loss in mitochondrial function due at least in part to the inhibition or inactivation of cytochrome c is a potentially fatal component of the apoptosis program of Jurkat cells. PMID- 8702952 TI - Replication protein A is a component of a complex that binds the human metallothionein IIA gene transcription start site. AB - Previous studies revealed that sequences surrounding the initiation sites in many mammalian and viral gene promoters, called initiator (Inr) elements, may be essential for promoter strength and for determining the actual transcription start sites. DNA sequences in the vicinity of the human metallothionein IIA (hMTIIA) gene transcription start site share homology with some of the previously identified Inr elements. However, in the present study we have found by in vitro transcription assays that the hMTIIA promoter does not contain a typical Inr. Electrophoretic mobility shift assays identified several DNA-protein complexes at the hMTIIA gene transcription start site. A partially purified protein fraction containing replication protein A (RPA) binds to the hMTIIA gene transcription start site and represses transcription from the hMTIIA promoter in vitro. In addition, overexpression of the human 70-kDa RPA-1 protein represses transcription of a reporter gene controlled by the hMTIIA promoter in vivo. These findings suggest that hMTIIA transcription initiation is controlled by a mechanism different from most mammalian and viral promoters and that the previously identified RPA may also be involved in transcription regulation. PMID- 8702953 TI - The UL8 subunit of the herpes simplex virus type-1 DNA helicase-primase optimizes utilization of DNA templates covered by the homologous single-strand DNA-binding protein ICP8. AB - The herpes simplex virus type-1 DNA helicase-primase is a heterotrimer encoded by the UL5, UL8, and UL52 genes. The core enzyme, specified by the UL5 and UL52 genes, retains DNA helicase, DNA-dependent nucleoside triphosphatase, and primase activities. The UL8 subunit has previously been implicated in increasing primer stability and in stimulating primer synthesis by the core enzyme. To further characterize the function of the UL8 subunit, we have examined its effect on the activities of the UL5/52 core enzyme using DNA templates covered by the herpes simplex virus type-1 single-strand DNA-binding protein ICP8. We found that while ICP8 stimulated the DNA helicase activity of the UL5/52 proteins up to 3-fold, maximum stimulation by ICP8 required the presence of UL8 protein. Moreover, UL8 protein was required to reverse the inhibitory effect of ICP8 on the DNA dependent ATPase and primase activities of the UL5/52 proteins. These observations were specific for ICP8 since the heterologous Escherichia coli single-strand DNA-binding protein could not substitute for ICP8. These data suggest that UL8 protein mediates an interaction between the UL5/52 core enzyme and ICP8 that optimizes the utilization of ICP8-covered DNA templates during DNA replication. PMID- 8702954 TI - Interaction of the fibrinolytic receptor, annexin II, with the endothelial cell surface. Essential role of endonexin repeat 2. AB - Endothelial cells express a cell surface co-receptor for plasminogen and tissue plasminogen activator (t-PA) which we recently identified as annexin II (Hajjar, K. A., Jacovina, A. T., and Chacko, J. (1994) J. Biol. Chem. 269, 21191-21197). This protein enhances the catalytic efficiency of t-PA-dependent plasmin generation by 60-fold (Cesarman, G. M., Guevara, C. A., and Hajjar, K. A. (1994) J. Biol. Chem. 269, 21198-21203). Here, we demonstrate that annexin II is constitutively translocated to the endothelial cell surface within 16 h of biosynthesis, and that cell surface annexin II comprises 4.3 +/- 1.0% of the total cellular pool. Exogenous 125I-annexin II bound to EGTA-washed endothelial cells with high affinity (Kd 49 nM) and in a calcium-dependent (I50 = 3 microM), phospholipid-sensitive manner. Peptides KASMKGLGTDED and YDSMKGKGTRDK, mimicking the calcium-binding "endonexin" motif (KGXGT) of annexin II, blocked its interaction with endothelial cells. Recombinant annexin II, bearing the calcium binding site substitution D161A of core repeat 2, failed to compete with binding of the wild type protein to the cell surface, while E246A and D321A mutants, corresponding to core repeats 3 and 4, behaved as effective competitors. These data suggest that translocated annexin II interacts with cell surface phospholipid via a high affinity calcium-dependent binding site that includes residues 118-122 (KGLGT) and the coordinating Asp161 of core repeat 2. Thus, calcium-regulated expression of annexin II on the endothelial cell surface may play a central role in control of plasmin-mediated processes. PMID- 8702955 TI - Sequences outside the homeodomain of bicoid are required for protein-protein interaction. AB - The Drosophila morphogenetic protein Bicoid (Bcd) is required for the development of anterior structures of the embryo. Bcd, a homeodomain protein, is distributed as an anterior-to-posterior gradient in the embryo. It stimulates the expression of the hunchback (hb) gene in the anterior half in an all-or-none fashion. We have recently shown that Bcd binds cooperatively to a hb enhancer element and proposed that cooperative DNA binding is facilitated by an interaction between Bcd molecules. In this report, we further analyze the interaction between Bcd molecules and define regions important for protein-protein interaction. We show that the homeodomain of Bcd alone fails to interact with another Bcd molecule efficiently. The protein sequence flanking either side of the homeodomain restores the protein-protein interaction function. Mutations in the homeodomain that affect DNA binding do not adversely affect the protein-protein interaction function, suggesting that the surfaces for DNA binding and protein-protein interaction are separable. Finally, we demonstrate that the homeodomain of Bcd alone, unlike the intact Bcd, fails to bind DNA cooperatively. These results further support the notion that cooperative DNA binding is facilitated by the interaction between Bcd molecules. They strongly suggest that protein-protein interaction is an important property of Bcd for its biological activities. PMID- 8702956 TI - Protein kinase C isozymes differentially regulate promoters containing PEA-3/12-O tetradecanoylphorbol-13-acetate response element motifs. AB - To investigate the regulation of promoters containing classical phorbol ester response sequences (PEA-3/12-O-tetradecanoylphorbol-13-acetate response element motifs) by protein kinase C (PKC) isozymes, co-transfections were performed in human dermal fibroblasts with a plasmid containing either the human collagenase promoter or the porcine urokinase plasminogen activator (uPA) promoter linked to the chloramphenicol acetyltransferase gene and a plasmid expressing an individual PKC isozyme. Using this experimental design, seven PKC isozymes were analyzed for their ability to trans-activate the collagenase and uPA promoters. Our results demonstrate that only PKC delta, epsilon, and eta trans-activated the collagenase promoter and that binding of Ap-1 family members to the collagenase 12-O tetradecanoylphorbol-13-acetate response element (TRE) was not responsible for the isozyme-specific trans-activation. In contrast, the uPA promoter was stimulated by all of the PKC isozymes examined (PKC alpha, betaII, gamma, delta, epsilon, zeta, and eta). These results indicate that PKC isozymes differentially regulate promoters containing PEA-3/TRE motifs and suggest that individual isozymes play unique roles within the cell. PMID- 8702957 TI - Dopamine transporter ligand binding domains. Structural and functional properties revealed by limited proteolysis. AB - Dopamine transporters (DATs) are members of the Na+- and Cl--dependent neurotransmitter and amino acid transporter family predicted by hydrophobicity analysis to have 12 transmembrane-spanning helices. The structure of DAT was studied using the photoaffinity compounds [125I]1-[2-(diphenylmethoxy)-ethyl]-4 [2-(4-azido-3-iodophenyl) ethyl] piperazine ([125I]DEEP), a 1-(2-diphenylmethoxy) ethyl-4-(3-phenyl propyl)piperazine (GBR analog), and [125I]-3beta-(p chlorophenyl)tropane-2beta-carboxylic acid, 4'-azido-3'-iodophenylethyl ester ([125I]RTI 82), a cocaine analog, which had been shown in a previous study to become incorporated into different regions of the DAT primary sequence. The proximity of the photolabeled binding sites to integral membrane structures was investigated by subjecting photolabeled membrane suspensions to limited proteolysis with trypsin and separately analyzing the resulting membranes and supernatants for the presence of photolabeled DAT fragments. Trypsin treatment of [125I] DEEP-labeled membranes generated labeled 45- and 14-kDa DAT fragments that immunoprecipitated with an epitope-specific antiserum generated against amino acids 42-59 near the first putative transmembrane domain, whereas [125I]RTI 82 was found in 32- and 16-kDa tryptic fragments that precipitated with an antiserum directed against a sequence near transmembrane domain 4 (amino acids 225-238). All of the photolabeled fragments were recovered in the protease-treated membranes, indicating that they possess integral membrane structures that prevent their release from the membrane as soluble forms. The size of the two smallest fragments in conjunction with their retention in the membrane suggests that incorporation of the photoaffinity ligands occurs in or near membrane spanning regions and delineates the maximum possible distance between the transmembrane structures, incorporated photolabel, and antibody epitopes. Carbohydrate analysis of the fragments identified sialic acids and N-linked oligosaccharides exclusively on the 45-kDa [125I]DEEP-labeled fragment, which, based on size, would be expected to contain four consensus glycosylation sites between putative transmembrane domains 3 and 4. Photoaffinity labeling after trypsin treatment of membranes showed that the larger but not the smaller fragments retain binding capacity, as the 45- and 32-kDa fragments were capable of becoming photolabeled. Binding of photoaffinity ligands at these fragments was displaced with the same pharmacology as that of intact DATs. These results verify numerous aspects of DAT structure and topology heretofore only predicted from theoretical considerations and extend our knowledge of DAT structure-function properties. PMID- 8702958 TI - The aurea and yellow-green-2 mutants of tomato are deficient in phytochrome chromophore synthesis. AB - The phytochrome-deficient aurea mutant of tomato has been widely used for the study of both phytochrome function and the role of other photoreceptors in the control of development in higher plants. To date the exact nature of the aurea mutation has remained unknown, though this information is clearly important for the interpretation of these studies. It has been proposed that aurea and yellow green-2, another mutant of tomato that has a similar phenotype to aurea, could be deficient in phytochrome chromophore synthesis. We have examined this hypothesis by measuring the activity of the enzymes committed to phytochrome chromophore synthesis in these mutants. The approach takes advantage of a recently developed high pressure liquid chromatography-based assay for the synthesis of the free phytochrome chromophore, phytochromobilin from its immediate precursors biliverdin IXalpha and heme. Isolated etioplasts from aurea and yellow-green-2 seedlings were specifically unable to convert biliverdin IXalpha to 3Z phytochromobilin and heme to biliverdin IXalpha, respectively. In addition, the level of total noncovalently bound heme in the mutants was the same as in wild type seedlings. Together, these results identify both aurea and yellow-green-2 as mutants that are deficient in phytochrome chromophore synthesis. PMID- 8702959 TI - Characterization of the interaction of FKBP12 with the transforming growth factor beta type I receptor in vivo. AB - The type I transforming growth factor-beta receptor (TbetaR-I) is the efferent component of the receptor complex, which presumably phosphorylates intracellular targets. FKBP12, a binding protein for FK506 and rapamycin, is shown to associate with the cytoplasmic region of TbetaR-I in vitro. In this report, we investigated the interaction of FKBP12 with TbetaR-I in vivo. FKBP12 interacts with TbetaR-I in mammalian cells as well as in yeast. Ligand addition does not affect the interaction, and both constitutively active and kinase-negative mutants of TbetaR I bind FKBP12. FKBP12 dissociates from TbetaR-I in the presence of a high concentration of FK506. The juxtamembrane region of TbetaR-I, containing the major phosphorylation sites by the type II receptor, is required for the interaction. One of the deletion mutants in this region, which was shown to mediate transcriptional response, does not bind FKBP12, suggesting that FKBP12 is not directly involved in TGF-beta signaling. Furthermore TbetaR-I does not phosphorylate FKBP12 in vitro. FKBP12 may not be a direct substrate of TbetaR-I but possibly modulates the TbetaR-I function through its interaction with the regulatory domain of the kinase. PMID- 8702960 TI - Involvement of the small GTPase rho in integrin-mediated activation of mitogen activated protein kinase. AB - Engagement and clustering of integrins triggers a number of intracellular signaling events, including activation of the mitogen-activated protein (MAP) kinases Erk1 and Erk2. To investigate the mechanism by which integrins mediate the activation of MAP kinases upon binding of NIH 3T3 cells to fibronectin, we assessed the effects of both inhibiting and activating the small GTPase Rho. We observed that inhibition of Rho by the Rho-specific inhibitor C3 exoenzyme or by a dominant negative Rho A (RhoN19) inhibited MAP kinase activation. Conversely, activation of Rho by expression of an activated Rho A mutant (RhoQ63L), or the Rho-specific guanine nucleotide exchange factor lbc, enhanced and partially mimicked activation of Erk2 by plating on fibronectin. These results therefore show that Rho is involved in the integrin-dependent activation of MAP kinase. PMID- 8702961 TI - Sequence determination of an extremely acidic rat dentin phosphoprotein. AB - The mineralization process associated with the conversion of predentin to dentin is believed to be initiated and controlled by a set of acidic regulatory noncollagenous proteins (NCPs) which include phosphophoryn, the major NCP in dentin. Phosphophoryn binds tightly to collagen and is believed to initiate the formation of apatite crystals which play a central role in the mineralization process. During the process of analyzing the 3' end of an odontoblast-specific cDNA which codes for dentin sialoprotein (Ritchie, H. H., Hou, H., Veis, A., and Butler, W. T. (1994) J. Biol. Chem. 269, 3698-3702), we discovered a 801-base pair open reading frame. This downstream open reading frame encodes a putative leader sequence and a very acidic mature protein sequence having a deduced amino acid composition containing high percentages of both Ser (43%) and Asp (31%) residues which closely coincides with the amino acid composition of phosphophoryns from human, bovine, rat, and rabbit (i. e. Asp (30-40%) and Ser (38-50%)). This newly identified cDNA therefore encodes a protein with characteristics similar to phosphophoryn. Here we present the cDNA sequence, the deduced amino acid sequence, and the prospective Ser residue-specific casein kinase I and II phosphorylation sites for this putative phosphophoryn. PMID- 8702962 TI - An interleukin-1beta converting enzyme-like protease is a key component of Fas mediated apoptosis. AB - Cytotoxic T lymphocytes (CTLs) are able to kill target cells bearing foreign antigen through two distinct mechanisms: granule- and Fas-mediated cytotoxicity. The exact events involved in the induction of target cell apoptosis remain elusive, but research indicates a role for members of the interleukin-1beta converting enzyme (ICE)/Ced-3 family of cysteine proteases. The exact nature of the protease(s) involved is yet to be determined. Here we use activity assays and peptide inhibitors of ICE/Ced-3 proteases to study their role in Fas-mediated killing. We find that while certain inhibitors block DNA fragmentation and chromium release, others do not. Most notably, potent inhibitors of CPP32 and ICE could not inhibit DNA fragmentation during all cases of Fas-mediated cytotoxicity although an "ICE" inhibitor could suppress 51Cr release. Additionally, we find that CPP32 is not cleaved in all target cells during Fas killing. Although ICE activity (as measured by a fluorogenic substrate) is present in cell lysates from anti-Fas-treated cells, we found no pro-IL-1beta-cleaving activity in these lysates. Taken together, our results suggest that an alternate pathway to DNA fragmentation exists, which does not involve CPP32 activity, and that CPP32 and ICE activities are not essential to Fas-mediated killing. PMID- 8702963 TI - The insulin-like growth factor II/mannose 6-phosphate receptor utilizes the same membrane compartments as GLUT4 for insulin-dependent trafficking to and from the rat adipocyte cell surface. AB - The insulin-like growth factor II (IGF-II)/mannose 6-phosphate (Man-6-P) receptor recycles in adipose cells between the cell surface and an intracellular storage pool, and the rate of this trafficking is markedly enhanced by insulin. We show here that the IGF-II/Man-6-P receptor is a constituent of the GLUT4-containing compartment ("GLUT4 vesicles") where it represents gp230, a major recycling protein detected earlier by cell surface biotinylation (Kandror, K. V., and Pilch, P. F. (1994) J. Biol. Chem. 269, 138-142). The GLUT4 vesicles include 10 15% of the total and all of the acutely insulin-responsive recycling population of the IGF-II/Man-6-P receptor. The main part of the IGF-II/Man-6-P receptor population is excluded from the pathway of GLUT4 trafficking and either resides permanently in intracellular membranes or has a much slower rate of cycling to the cell surface. Thus, GLUT4 vesicles mediate the insulin-dependent delivery to the cell surface of the IGF-II/Man-6-P receptor as well as the other recyclable proteins with extracellular functional domains (GLUT4 and the aminopeptidase gp160). PMID- 8702964 TI - Cleavage of CPP32 by granzyme B represents a critical role for granzyme B in the induction of target cell DNA fragmentation. AB - Cytotoxic T lymphocytes (CTLs) are able to recognize and destroy target cells bearing foreign antigen using one of two distinct mechanisms: granule- or Fas mediated cytotoxicity. The exact mechanisms involved in the induction of apoptotic cell death remain elusive; however, it seems likely that a family of cysteine proteases related to interleukin-1beta converting enzyme are involved. One family member, CPP32, has been identified as an intracellular substrate for granzyme B, a CTL-specific serine protease responsible for the early induction of target cell DNA fragmentation. Here we use cytolytic cells from granzyme B deficient mice to confirm that cleavage and activation of CPP32 represents a nonredundant role for granzyme B and that this activation plays a role in the induction of DNA fragmentation in target cells, a signature event for apoptotic cell death. A peptide inhibitor of CPP32-like proteases confirmed the function of these enzymes in fragmentation. 51Cr release was not suppressed under these conditions, suggesting that granzyme B cleavage of CPP32 is primarily involved in the induction of DNA fragmentation and not membrane damage during CTL-induced apoptosis. PMID- 8702965 TI - Purine-specific nucleoside N-ribohydrolase from Trypanosoma brucei brucei. Purification, specificity, and kinetic mechanism. AB - Trypanosomes have no de novo purine biosynthesis and thus depend upon salvage pathways to obtain purines for their metabolic pathways and for the biosynthesis of nucleic acids. An inosine-adenosine-guanosine preferring nucleoside hydrolase (IAG-nucleoside hydrolase) from the African trypanosome Trypanosoma brucei brucei represents approximately 0.2% of the soluble protein in this organism. The enzyme has been purified over 400-fold to >95% homogeneity from the bloodstream form of this parasite. IAG-nucleoside hydrolase is a dimer of Mr 36,000 subunits. The kcat/Km for inosine, adenosine, and guanosine are 1.9 x 10(6), 1.2 x 10(6), and 0.83 x 10(6) M -1 s-1, respectively. The kinetic mechanism with inosine as substrate is rapid equilibrium with random product release. The turnover rate for inosine at 30 degrees C is 34 s-1. Pyrimidine nucleosides are poor substrates with kcat/Km values of approximately 10(3) M -1 s-1. Deoxynucleosides are also poor substrates with kcat/Km values near 10(2) M -1 s-1. AMP is not a detectable substrate and there is no measurable purine nucleoside phosphorylase activity. 3 Deazaadenosine, 7-deazaadenosine (tubercidin), and formycin B are competitive inhibitors with Kis of 1.8, 59, and 13 microM, respectively. The Km shows a slight dependence on pH with a pH optimum around 7. The Vmax/Km data indicate there are two ionizable enzymatic groups, pKa 8.6, required for the formation of the Michaelis complex. The Vmax data indicate three ionizable groups involved in catalysis. Two essential groups exhibit pKa values of 8.8, and a third group with a pKa of 6.5 increases the Vmax an additional 10-fold. All three groups must be protonated for optimum catalytic activity. PMID- 8702966 TI - Arachidonic acid activates the noncapacitative entry of Ca2+ during [Ca2+]i oscillations. AB - Current models for agonist-activated Ca2+ entry in nonexcitable cells focus on the capacitative mechanism where entry is activated as a downstream result of the sustained depletion of agonist-sensitive stores without any direct requirement for inositol phosphates. This mechanism has been shown to be important for the sustained Ca2+ signals seen in a variety of nonexcitable cells under conditions of maximal stimulation. In contrast, relatively little attention has been given to Ca2+ entry under more physiological levels of agonist where, for example, oscillating Ca2+ responses are common. In recent studies using cells from the exocrine avian nasal gland, we have shown that agonist-activated Ca2+ entry under these conditions demonstrates properties that are inconsistent with current versions of the capacitative model. We now report that activation of this novel noncapacitative Ca2+ entry is via a distinct signaling pathway involving an agonist-induced, phospholipase A2-mediated generation of arachidonic acid. PMID- 8702967 TI - Phospholamban regulates the Ca2+-ATPase through intramembrane interactions. AB - There is clear evidence for direct regulatory protein-protein interactions between phospholamban (PLN) and the Ca2+-ATPase of cardiac sarcoplasmic reticulum (SERCA2a) in cytoplasmic domains, but there is less clear evidence for regulatory interactions in the transmembrane domains of the two proteins. We have now coexpressed SERCA isoforms with the transmembrane sequence of PLN and with epitope-tagged transmembrane sequences of PLN to study intramembrane interactions in the absence of cytoplasmic interactions. Coexpression of the transmembrane sequence of phospholamban (Met-PLN28-52) with SERCA1a, SERCA2a, and SERCA3 inhibited Ca2+ transport by lowering apparent Ca2+ affinity. Addition of the hemagglutinin (HA) epitope to the transmembrane sequence of PLN (HA-PLN28-52) or deletion of PLN residues 21-29 (PLN1-20-PLN30-52) "supershifted" apparent Ca2+ affinity to values lower than those observed with native PLN without uncoupling Ca2+ transport from ATP hydrolysis. Inhibition by PLN1-20-PLN30-52 or by Flag PLN28-52 was reversed by PLN antibody or by Flag antibody, demonstrating that inhibition by these constructs is reversible and that the inhibitory constructs are properly oriented in the membrane. These results suggest that PLN modulates the apparent Ca2+ affinity of SERCA2a through intramembrane interactions, which are disrupted at long range and in concert with disruption of the well characterized cytoplasmic interactions. PMID- 8702968 TI - Identification of a putative effector for Cdc42Hs with high sequence similarity to the RasGAP-related protein IQGAP1 and a Cdc42Hs binding partner with similarity to IQGAP2. AB - Cdc42 is a Ras-related GTP-binding protein that has been implicated in the regulation of the actin cytoskeleton and cell morphology. In this study, we have identified a protein with a molecular mass approximately 180 kDa from rabbit liver cytosol (designated p180), which binds preferentially to the GTP- and guanosine 5'-3-O-(thio)triphosphate-bound forms of Cdc42. Binding of p180 to GTP bound Cdc42 maintains it in the GTP-bound state. Another cytosolic protein, with an apparent molecular mass of 175 kDa (p175), was also found to interact with Cdc42, but this association showed less dependence on guanine nucleotides. Both p180 and p175 were capable of binding to Rac1 but not to RhoA or Ha-Ras. The limit functional domain of the Cdc42-GAP protein did not compete with p180 or p175 for binding to Cdc42. However, the Cdc42-binding domain from mPAK-3, a member of the PAK (p21 activated kinase) family of serine/threonine kinases, competed with both proteins. The binding of p180 or p175 was inhibited by mutations of the putative effector loop of Cdc42. p180 and p175 also bound less effectively to a Cdc42/Ras chimera in which loop 8 from Ras was substituted for the predicted loop 8 in Cdc42 that includes a 13-amino acid insert present in all Rho family members but absent in Ras. Microsequencing of a p180 peptide revealed 92% identity with the human IQGAP1 protein, while two peptides derived from p175 were 89 and 100% identical to human IQGAP2. These findings identify IQGAP1 and IQGAP2 as a new class of target/effectors that utilize both regions of the switch I domain and an insert region distinct to Rho proteins for binding to Cdc42. PMID- 8702969 TI - Metabolic processing of gangliosides by normal and Salla human fibroblasts in culture. A study performed by administering radioactive GM3 ganglioside. AB - Cultured fibroblasts from normal subjects and from subjects affected by Salla disease, characterized by the lack or misfunction of the membrane carrier responsible for the egress of sialic acid from lysosomes, were fed with ganglioside GM3 labeled at the sialic acid acetyl group, [Neu5Ac-3H]GM3, or at C 3 of sphingosine (Sph), [Sph-3H]GM3, or at C-1 of stearoyl chain, [stearoyl 14C]GM3. After a 15-h pulse the total amount of cell-bound GM3 corresponded to about 2% of the endogenous ganglioside content. Cells were then subjected to a 72 h chase, and the radioactive products from both ganglioside catabolism and salvage processes of catabolic fragments were measured. These data indicated that about 50% of the cell-bound ganglioside underwent metabolic processing, suggesting a ganglioside half-life of 2-3 days. [Neu5Ac-3H] formed from [Neu5Ac 3H]GM3 degradation was mostly re-cycled for the biosynthesis of gangliosides and sialoglycoproteins, only a minor part being degraded to [3H]water, which constituted only 1.6% of total metabolite linked radioactivity. [Sph-3H] from the [Sph-3H]GM3 degradation was partly re-cycled for the biosynthesis of gangliosides, neutral glycosphingolipids and sphingomyelin, and partly (about 20% of the total metabolite linked radioactivity) degraded to [3H]water. In Salla fibroblasts metabolic processing of [Neu5Ac-3H]GM3 produced large amounts of free [3H]Neu5Ac, and a reduced incorporation of radioactivity into glycoconjugates (as compared to normal cells). However, the accumulation of free Neu5Ac was not accompanied by an increase of tritiated water. LacCer and Cer from [stearoyl 14C]GM3 catabolism were found to accumulate in Salla fibroblasts, an indication that the enzymes of glycosphingolipid metabolism were affected by the impairment of Neu5Ac egress from lysosomes. Particularly relevant was the accumulation of ceramide which was hardly detectable in control cells. PMID- 8702970 TI - An allosteric Ca2+ binding site on the beta3-integrins that regulates the dissociation rate for RGD ligands. AB - Here we use a model RGD-containing ligand to study how Ca2+ and Mg2+ regulate ligand binding to beta3-integrins. Fab-9, an antibody that contains an optimized RGD loop in its antigen binding site (Barbas, C. F., Languino, L., and Smith, J. W. (1993) Proc. Natl. Acad. Sci. U. S. A. 90, 10003-10007), was used as the model ligand. Across a physiologic range of Mg2+, Fab-9 bound to both alphavbeta3 and alphaIIbbeta3 with a monophasic binding isotherm. Across the same range of Ca2+, the binding of Fab-9 to the beta3-integrins was biphasic. Low concentrations of Ca2+ (microM) promoted the binding of Fab-9. Higher concentrations of Ca2+ (m) blocked Fab-9 binding. These data suggest that Ca2+ binds to two distinct classes of sites on the beta3-integrins, with the low affinity Ca2+ binding site(s) being an inhibitory site. We designate this inhibitory site(s) as the I site. Further biochemical characterization showed that the I site has the following characteristics: 1) it is specific for Ca2+; 2) it is allosteric to the ligand binding site; 3) its occupation increases the dissociation rate between integrin and RGD ligand; and 4) occupation of the I site can induce cellular deadhesion. PMID- 8702971 TI - Substrate recognition by tissue factor-factor VIIa. Evidence for interaction of residues Lys165 and Lys166 of tissue factor with the 4-carboxyglutamate-rich domain of factor X. AB - Tissue factor (TF) is the protein cofactor for factor VIIa (FVIIa), the first serine protease of the clotting cascade. Previous studies using alanine mutagenesis have identified TF residues Lys165 and Lys166 as important for factor X (FX) activation, hypothesizing either that these residues interact with phospholipid head groups or that they directly or indirectly promote macromolecular substrate binding. In the recently reported x-ray crystal structure of the isolated extracellular domain of TF, both Lys165 and Lys166 are solvent-exposed and predicted to be near the phospholipid surface in intact TF. We hypothesized that these residues may in fact be ideally positioned to interact with the 4-carboxyglutamate-rich domain (Gla domain) of FX. We therefore predicted that mutations at Lys165 and Lys166 should have no effect on the activation of Gla domainless FX. To test this hypothesis, we mutated both residues Lys165 and Lys166 of TF to Ala, Glu, or Gln and examined the ability of these double mutants to support FVIIa-mediated activation of FX, Gla domainless FX, and factor IX (FIX). Each TF mutant was equivalent to wild-type TF in both FVIIa binding and promotion of FVIIa amidolytic activity. However, all three mutants were markedly deficient in supporting FIX and FX activation, with FX activation rates decreased more than FIX activation rates. In both reactions, the TF mutants exhibited different extents of activity: Gln165-Gln166 > Ala165-Ala166 > Glu165-Glu166. In sharp contrast, all three TF mutants were equivalent to wild type TF in supporting activation of Gla domainless FX by FVIIa. Interestingly, the deficiency of the mutants in FX activation was less pronounced when Gla domainless FVIIa was used in place of native FVIIa. Together, these findings suggest that TF residues Lys165 and Lys166 contribute to a binding site for the Gla domain of FX (and perhaps other substrates) and that this interaction may be facilitated by the presence of the Gla domain of FVIIa. PMID- 8702972 TI - Chimeric granulocyte/macrophage colony-stimulating factor/transforming growth factor-beta (TGF-beta) receptors define a model system for investigating the role of homomeric and heteromeric receptors in TGF-beta signaling. AB - Transforming growth factor-beta (TGF-beta) belongs to a family of ligands that regulate cell growth and differentiation. The most commonly observed receptors are referred to as the type I, type II, and type III (betaglycan) TGF-beta receptors. Two receptor models have been presented to account for the various cellular responses to TGF-beta. The first proposes that all TGF-beta signaling results from the formation of a heteromeric type I/type II complex, while the second suggests that distinct type I or type II TGF-beta receptor combinations mediate aspects of TGF-beta signaling. We have addressed this general question relating to TGF-beta signaling by constructing chimeric receptors consisting of the extracellular domain of the granulocyte/macrophage colony-stimulating factor (GM-CSF) alpha or beta receptor fused to the transmembrane and cytoplasmic domain of the type I or type II TGF-beta receptor. Since high affinity GM-CSF binding requires dimerization of the alpha and beta ligand binding subunits, the response elicited by defined type I and/or type II TGF-beta receptor cytoplasmic domain homomers or heteromers can be examined. We show in mesenchymal AKR-2B cells that while TGF-beta-dependent transient luciferase activity, endogenous gene activity, and long-term biological responses are similarly induced by activating the chimeric heteromeric receptors with GM-CSF as the endogenous TGF-beta receptor, chimeric homomeric type I/type I or type II/type II receptors are signaling incompetent. PMID- 8702973 TI - Intracellular distribution of Arf proteins in mammalian cells. Arf6 is uniquely localized to the plasma membrane. AB - Subcellular distributions of the five human Arf proteins were examined, using a set of isoform-specific polyclonal and a pan-Arf monoclonal antibodies. Subcellular fractionation of cultured mammalian cells allowed the demonstration that Arf6 is uniquely localized to the plasma membranes of Chinese hamster ovary cells. The plasma membrane distrubution was unaffected by either GTPgammaS (guanosine 5'-O-(3-thio)triphosphate) or brefeldin A, an activator and inhibitor of Arf activities, respectively. In contrast, Arf proteins 1, 3, 4, and 5 were predominantly cytosolic but could be recruited to a variety of intracellular membranes, but not plasma membranes, upon incubation in the presence of GTPgammaS. The GTPgammaS-promoted binding of the cytosolic Arf proteins to membranes was blocked by brefeldin A. The stable association of Arf6 with plasma membranes and the insensitivity of its localization to either GTPgammaS or brefeldin A revealed a clear distinction between Arf6 and the other Arf isoforms. Localization of Arf6 to the plasma membrane suggests a unique cellular role for this isoform at the plasma membrane, but failure to find endogenous Arf6 on endocytic structures, including clathrin-coated vesicles, appears inconsistent with the proposed role of Arf6 in assembly of coat structures or endosomes in transfected fibroblasts (1,2). PMID- 8702974 TI - A novel family of developmentally regulated mammalian transcription factors containing the TEA/ATTS DNA binding domain. AB - We describe the molecular cloning of two novel human and murine transcription factors containing the TEA/ATTS DNA binding domain and related to transcriptional enhancer factor-1 (TEF-1). These factors bind to the consensus TEA/ATTS cognate binding site exemplified by the GT-IIC and Sph enhansons of the SV40 enhancer but differ in their ability to bind cooperatively to tandemly repeated sites. The human TEFs are differentially expressed in cultured cell lines and the mouse (m)TEFs are differentially expressed in embryonic and extra-embryonic tissues in early post-implantation embryos. Strikingly, at later stages of embryogenesis, mTEF-3 is specifically expressed in skeletal muscle precursors, whereas mTEF-1 is expressed not only in developing skeletal muscle but also in the myocardium. Together with previous data, these results point to important, partially redundant, roles for these TEF proteins in myogenesis and cardiogenesis. In addition, mTEF-1 is strongly coexpressed with mTEF-4 in mitotic neuroblasts, while accentuated mTEF-4 expression is also observed in the gut and the nephrogenic region of the kidney. These observations suggest additional roles for the TEF proteins in central nervous system development and organogenesis. PMID- 8702975 TI - Human cancer cells exhibit protein kinase C-dependent c-erbB-2 transmodulation that correlates with phosphatase sensitivity and kinase activity. AB - The c-erbB-2 receptor tyrosine kinase is often overexpressed in human tumors, but the functional implications of this phenotype remain unclear. We previously used phosphorylation-specific antibodies to define major differences in c-erbB-2 tyrosine kinase activity between overexpressing human tumor cell lines (Epstein, R. J., Druker, B. J., Roberts, T. M., and Stiles, C. D. (1992) Proc. Natl. Acad. Sci. U. S. A. 89, 10435-10439). Here we extend this approach to define the relationship between c-erbB-2 tyrosine phosphorylation and protein kinase C (PKC) dependent transmodulation. Phosphorylation-specific antibodies to the juxtamembrane PKC site Thr686 recognize tyrosine-dephosphorylated wild-type c erbB-2 following G8/DHFR 3T3 cell treatment with PKC agonists. B104-1-1 cells transformed by activated c-erbB-2 express a subset of tyrosine-phosphorylated receptors that are homologously phosphorylated on Thr686, indicating that Thr686 phosphorylation alone is insufficient to abrogate receptor tyrosine phosphorylation. Similarly, the c-erbB-2-overexpressing human cancer cell lines SK-Ov-3 and BT-474 express constitutively Thr686-phosphorylated receptors. SK-Ov 3 cells express predominantly kinase-inactive c-erbB-2 that is heavily Thr686 phosphorylated, indicating that Thr686 phosphorylation in this line is heterologous in origin. In contrast, BT-474 cells express constitutively autophosphorylated c-erbB-2 despite Thr686 phosphorylation. These results indicate that Thr686 phosphorylation does not directly abolish c-erbB-2 activity and suggest that such phosphorylation reflects constitutive PKC activity induced by either receptor-activating mutations or heterologous growth factors. The latter possibility suggests in turn that c-erbB-2 interacts in an as yet undefined way with heterologous growth factor receptors in human tumor cells. PMID- 8702976 TI - Ultraviolet B and H2O2 are potent inducers of vascular endothelial growth factor expression in cultured keratinocytes. AB - Vascular endothelial growth factor (VEGF), also known as vascular permeability factor, is strongly expressed by epidermal keratinocytes during wound healing, in psoriasis, and in bullous diseases such as erythema multiforme and bullous pemphigoid. All of these disorders are characterized by increased microvascular permeability and angiogenesis. Since the development of erythema as a result of hyperpermeable blood vessels is also a common feature after excess sun exposure, we speculated about an up-regulation of VEGF expression by ultraviolet (UV) light. To test this hypothesis, we analyzed the effect of UVB irradiation on VEGF expression in cultured keratinocytes. Thereby we found a large increase in VEGF mRNA and protein levels upon irradiation of quiescent keratinocytes with sublethal and physiologically relevant doses of UVB. Although H2O2 was also a potent inducer of VEGF expression, the effect of UVB irradiation is unlikely to be mediated by reactive oxygen species as determined by the use of antioxidants. Further experiments revealed that the UVB-induced overexpression of VEGF is dependent on de novo protein synthesis and might occur via release of soluble mediators, which subsequently turn on VEGF expression. In summary, our results suggest a novel role of VEGF in the induction of erythema after excess sun exposure. PMID- 8702977 TI - Sensitization of the HIV-1-LTR upon long term low dose oxidative stress. AB - Human lymphoid cell lines were transfected with HIV-1-LTR-CAT DNA and permanently transformed cell lines were obtained. These transformed cell lines were treated with 0.01 mM H2O2 for 25 days and the chloramphenicol acetyltransferase (CAT) activities of these cell lines were measured. The CAT activities of transformed cell lines were latent in normal culture conditions, but were activated by retreatment with 0.2 mM H2O2 for 1 h. On treatment with 0.05 mM H2O2 for 1 h, the CAT activity of these cell lines maintained in normal conditions remained latent, whereas cell lines pretreated with 0.01 mM H2O2 for 25 days were greatly activated by this treatment. Here, the HIV-1 promoter seemed latent in normal culture conditions, but it could be activated by a comparatively low concentration (0.05 mM) of H2O2 after treatment with a dilute H2O2 (0.01 mM) for about 1 month. Many patients infected with human immunodeficiency virus 1 (HIV-1) show a long latent period before development of AIDS. During this latent period, their infected cells may be subjected to oxidative stress due to metabolism and physical movement. The present results indicate that oxidative stress may cause activation of the HIV-1 promoter in patients with latent HIV-1. PMID- 8702978 TI - Isolation and sequencing of the rho gene from Streptomyces lividans ZX7 and characterization of the RNA-dependent NTPase activity of the overexpressed protein. AB - The gene for transcription termination factor Rho was isolated from Streptomyces lividans ZX7. It encoded a 77-kDa polypeptide (Rho 77) with considerable homology to known Rho factors. An atypical hydrophilic region of 228 residues was found within the N-terminal RNA-binding domain. Only Rho from Micrococcus luteus and Mycobacterium leprae (closely related GC-rich Gram-positive bacteria) had an analogous sequence. Rho 77 was overexpressed in Escherichia coli and purified using an N-terminal hexahistidine-tag. Rho 77 displayed a broad RNA-dependent ATPase activity, with poly(C) RNA being no more than 4-fold more effective than poly(A). This contrasts with the ATPase activity of Rho from E. coli which is stimulated primarily by poly(C) RNA. Rho 77 was a general RNA-dependent NTPase, apparent Km values for NTPs were: GTP 0.13 mM, ATP 0.17 mM, UTP 1.1 mM, and CTP >2 mM. Rho 77 poly(C)-dependent ATPase activity was inhibited by heparin, unlike the E. coli Rho. The antibiotic bicyclomycin inhibited the in vitro RNA-dependent ATPase activity of Rho 77, did not inhibit growth of streptomycetes but delayed the development of aerial mycelia. N-terminal deletion analysis to express a truncated form of Rho (Rho 72, 72 kDa) indicated that the first 42 residues of Rho 77 were not essential for RNA-dependent NTPase activity and were not the targets of inhibition by heparin or bicyclomycin. PMID- 8702980 TI - Differential regulation of G-protein-mediated signaling by chemokine receptors. AB - Monocyte chemoattractant protein-1 (MCP-1) is a member of a family of chemotactic cytokines that induce directed migration of leukocytes via activation of seven transmembrane domain receptors. To identify G-proteins that couple to the two forms of the MCP-1 receptor, as well as to related chemokine receptors, we have performed cotransfection experiments in mammalian cells. In COS-7 cells, the type A and type B MCP-1 receptors coupled to Galphai, Galphaq, and Galpha16, whereas the macrophage inflammatory protein-1alpha/RANTES (regulated on activation, normal T cell-expressed and secreted) receptor (C-CR1) coupled to Galphai and Galphaq but failed to couple to Galpha16. In HEK-293 cells, however, the MCP-1 receptors and C-CR1 coupled to Galphaq but failed to couple to Galpha16. In contrast, the interleukin-8 and C5a receptors did not couple to Galphaq in either COS-7 or HEK-293 cells but did couple to Galpha16. Exchange of intracellular loops between the MCP-1 and interleukin-8 receptors to create chimeric receptors revealed that the third loop of the MCP-1 receptor accounted for virtually all of the coupling to Galphaq. We conclude that the MCP-1 and related chemokine receptors couple to multiple G-proteins, that coupling is cell type-specific, and that the third intracellular loop of the C-C type receptors mediates Galphaq coupling. PMID- 8702979 TI - Innate immunity. Isolation of several cysteine-rich antimicrobial peptides from the blood of a mollusc, Mytilus edulis. AB - We have isolated from the blood of immune-challenged and untreated mussels (Mytilus edulis) antibacterial and antifungal peptides. We have characterized two isoforms of a novel 34-residue, cysteine-rich, peptide with potent bactericidal activity and partially characterized a novel 6.2-kDa antifungal peptide containing 12 cysteines. We report the presence of two members of the insect defensin family of antibacterial peptides and provide a phylogenetic analysis that indicates that mollusc and arthropod defensins have a common ancestry. Our data argue that circulating antimicrobial peptides represent an ancient host defense mechanism that predated the separation between molluscs and arthropods at the root of the Cambrian, about 545 million years ago. PMID- 8702981 TI - Quantitating direct chlorine transfer from enzyme to substrate in chloroperoxidase-catalyzed reactions. AB - Substrate competition methods that were previously used to quantitate the involvement of free Cl2 in the chloride-dependent peroxidatic reactions catalyzed by chloroperoxidase (CPO) (Libby, R. D., Shedd, A. L., Phipps, K. A., Beachy, T. M., and Gerstberger, S. M., (1992) J. Biol. Chem. 267, 1769-1775) are extended to CPO-catalyzed halogenation reactions. Relative substrate specificities of halogen acceptor substrates (RHs) antipyrine (ap), NADH, 2-chlorodimedone (2cd), and barbituric acid (ba) are compared with previously studied peroxidatic substrates catechol (cat) and 2, 4,6-trimethylphenol (tmp) in their reactions with the CPO H2O2-Cl system versus the hypochlorite-Cl system. Studies were carried out at pH 2.75 over a chloride concentration range of 1-100 mM and at pH 4.80 over a chloride concentration range of 100-400 mM. Competition studies involved successive pairwise comparisons of substrates of increasing enzyme specificity. The orders of specificities, ba > 2cd > ap > cat > tmp at pH 2.75 and ba > 2cd > NADH > ap > cat > tmp at pH 4.80, are the same for both the CPO-H2O2-Cl and hypochlorite-Cl systems. However, the magnitudes of the specificities are different between the two systems. In all comparisons except ap versus cat, the specificity of the CPO-H2O2-Cl system toward the preferred substrate is higher than that of the hypochlorite-Cl system. Quantitative comparisons between specificities of CPO-H2O2-Cl and hypochlorite-Cl systems indicate that at least 98% of the CPO-catalyzed halogenation reactions of ba, 2cd, NADH, and ap occur by mechanisms in which the substrate reacts directly with the enzyme. Thus, less than 2% of any of the CPO reactions could possibly involve a free oxidized halogen intermediate. All data are consistent with a mechanism in which RH binds to the CPO chlorinating intermediate (EOCl), and the chlorine atom is transferred directly from EOCl to RH. Further, the results indicate that any halogenation substrate with a higher CPO specificity than ap must also undergo direct chlorine transfer from the enzyme. These results underscore the critical need for quantitative kinetic evidence in establishing the extent of involvement of any potential reaction intermediate. Finally, this work calls into question the long held assumption of the obligatory involvement of hypochlorite as an intermediate in myeloperoxidase reactions. It supports the recent kinetic evidence presented by Marquet and Dunford for direct chlorine transfer in myeloperoxidase-catalyzed chlorination of tuarine (Marquet, L. A., and Dunford, H. B. (1994) J. Biol. Chem. 269, 7950-7956). PMID- 8702982 TI - Characterization of the primary sigma factor of Staphylococcus aureus. AB - RNA polymerase (RNAP) isolated from Staphylococcus aureus is deficient in sigma factor and is poorly active in transcription assays. Based on amino acid sequence homology of the Bacillus subtilis vegetative sigma factor sigmaA and the predicted product of the chromosomally located plaC gene of S. aureus, it was hypothesized that plaC could encode the vegetative sigma factor. We cloned plaC under a T7 promoter and overexpressed it in Escherichia coli strain BL21(DE3)pLysE. The overproduced protein, present in inclusion bodies, was solubilized with guanidine hydrochloride, renatured, and purified by DEAE Sephacel and Sephadex G-75 chromatography. The purified protein, designated sigmaSA, cross-reacted with the B. subtilis anti-sigmaA antibody. E. coli core RNAP, reconstituted with sigmaSA, initiated promoter-specific transcription from the S. aureus promoters hla, sea, and sec and from the E. coli promoters rpoH P1, rpoH P4, and ColE1 RNA-1, which are recognized by the E. coli sigma70. sigmaSA, when added to the purified RNAP from S. aureus, stimulated transcriptional activity of the RNAP up to 72-fold. As determined by primer extension studies, the 5'-ends of the sigmaSA-initiated mRNAs synthesized in vitro from the agr P2 and sea promoters are in general agreement with the 5'-ends of the cellular RNAs. Disruption of the plaC gene on the S. aureus chromosome was lethal. We conclude that plaC encodes the primary sigma factor in S. aureus. PMID- 8702983 TI - Identification of the cAMP response element that controls transcriptional activation of the insulin-like growth factor-I gene by prostaglandin E2 in osteoblasts. AB - Insulin-like growth factor-I (IGF-I), a multifunctional growth factor, plays a key role in skeletal growth and can enhance bone cell replication and differentiation. We previously showed that prostaglandin E2 (PGE2) and other agents that increase cAMP activated IGF-I gene transcription in primary rat osteoblast cultures through promoter 1 (P1), the major IGF-I promoter, and found that transcriptional induction was mediated by protein kinase A. We now have identified a short segment of P1 that is essential for full hormonal regulation and have characterized inducible DNA-protein interactions involving this site. Transient transfections of IGF-I P1 reporter genes into primary rat osteoblasts showed that the 328-base pair untranslated region of exon 1 was required for a full 5.3-fold response to PGE2; mutation in a previously footprinted site, HS3D (base pairs +193 to +215), reduced induction by 65%. PGE2 stimulated nuclear protein binding to HS3D. Binding, as determined by gel mobility shift assay, was not seen in nuclear extracts from untreated osteoblast cultures, was detected within 2 h of PGE2 treatment, and was maximal by 4 h. This DNA-protein interaction was not observed in cytoplasmic extracts from PGE2-treated cultures, indicating nuclear localization of the protein kinase A-activated factor(s). Activation of this factor was not blocked by cycloheximide (Chx), and Chx did not impair stimulation of IGF-I gene expression by PGE2. In contrast, binding to a consensus cAMP response element (CRE; 5'-TGACGTCA-3') from the rat somatostatin gene was not modulated by PGE2 or Chx. Competition gel mobility shift analysis using mutated DNA probes identified 5'-CGCAATCG-3' as the minimal sequence needed for inducible binding. All modified IGF-I P1 promoterreporter genes with mutations within this CRE sequence also showed a diminished functional response to PGE2. These results identify the CRE within the 5'-untranslated region of IGF I exon 1 that is required for hormonal activation of IGF-I gene transcription by cAMP in osteoblasts. PMID- 8702984 TI - A yeast transcriptional stimulatory protein similar to human PC4. AB - A yeast protein has been identified that stimulates basal transcription by RNA polymerase II, binds both single- and double-stranded DNA, and interacts with both a general transcription factor and a transcriptional activator. Phosphorylation appears to regulate these interactions. The gene for the transcriptional stimulatory protein, termed TSP1, was cloned and found to be dispensable for yeast cell viability. The deduced amino acid sequence is similar to that of mammalian coactivator protein PC4. PMID- 8702985 TI - Molecular studies of CtpA, the carboxyl-terminal processing protease for the D1 protein of the photosystem II reaction center in higher plants. AB - The D1 reaction center protein of the Photosystem II complex in green plants is synthesized with a short carboxyl-terminal extension. Proteolytic cleavage and removal of this extension peptide in the thylakoid lumen are necessary for the assembly of a manganese cluster that is essential for the oxygen evolution activity of Photosystem II. We have isolated cDNAs encoding CtpA, the carboxyl terminal processing protease for the D1 protein, from two higher plants, spinach and barley. In each of these organisms, CtpA is encoded by a single copy nuclear gene, and its steady-state mRNA levels are light-regulated. The CtpA protein is detectable in etiolated material, and its level increases approximately 5-fold upon illumination. Moreover, the CtpA gene is expressed in shoot tissues and not in roots. In its precursor form, the CtpA protein harbors a bipartite transit sequence characteristic for thylakoid lumenal proteins. Cell fractionation studies demonstrated that CtpA is associated with thylakoid membranes and is resistant to treatments with thermolysin, consistent with its localization in the lumen of thylakoids. Comparisons of the sequence of the higher plant CtpA enzyme with those of other related carboxyl-terminal processing proteases suggest that these proteins constitute a new family of proteases. PMID- 8702986 TI - Stability and oligomeric equilibria of refolded interleukin-1beta converting enzyme. AB - We report the preparation and characterization of interleukin-1beta converting enzyme (ICE) refolded from its p20 and p10 protein fragments. Refolded ICE heterodimer (p20p10) was catalytically active but unstable, and in size exclusion chromatography eluted at an apparent molecular mass of 30 kDa. The mechanisms of the observed instability were pH-dependent dissociation at low enzyme concentrations, and autolytic degradation of the p10 subunit at high concentrations. Binding and subsequent removal of a high affinity peptidic inhibitor increased the apparent molecular mass to 43 kDa (by size exclusion chromatography), and significantly increased its stability and specific activity. Chemical cross-linking and SDS-polyacrylamide gel electrophoresis analysis of the 43-kDa size exclusion chromatography conformer revealed a 60-kDa species, which was absent in the 30-kDa conformer, suggesting that inhibitor binding caused formation of a (p20p10)2 homodimer. The observation of a reversible equilibrium between ICE (p20p10) and (p20p10)2 suggests that analogous associations, possibly between ICE and ICE homologs, can occur in vivo, resulting in novel oligomeric protease species. PMID- 8702987 TI - Purification and properties of rat liver nuclear proteins that interact with the hepatitis B virus enhancer 1. AB - The hepatitis B virus enhancer 1 element plays a fundamental role in the liver specific regulation of hepatitis B virus gene expression. A central region of enhancer 1, the enhancer core domain, contains at least four cis-acting sequence motifs that are essential for enhancer 1 activity. In this study, we have investigated an essential motif within the core domain previously defined as footprint V (FPV). Transient transfection analyses demonstrate that FPV is capable of independently functioning in a liver-specific manner to activate transcription. Therefore, to further examine the liver-specific properties of FPV mediated enhancer 1 activity, we have carried out the biochemical purification and characterization of FPV binding activity from rat liver nuclei. This study has conclusively identified hepatocyte nuclear factor 3beta (HNF-3beta), a liver enriched member of the HNF-3/forkhead gene family, as the predominant purified protein that interacts with the FPV motif. Moreover, a cellular protein(s) that copurified with HNF-3beta specifically interacts with a novel sequence motif that partially overlaps FPV. Since this novel motif contains a palindromic sequence, we have tentatively referred to the protein(s) that binds to this site as palindrome-binding factor (PBF). Additional evidence indicates that HNF-3beta and PBF cooperatively interact with enhancer 1. Therefore, this study supports the hypothesis that FPV-mediated enhancer activity involves a cooperative interplay between HNF-3beta and at least one other enhancer 1-binding protein, PBF. PMID- 8702988 TI - Domain-specific gene activation by parathyroid hormone in osteoblastic ROS17/2.8 cells. AB - Parathyroid hormone (PTH)-mediated gene activation was assessed in the osteoblast like rat cell line ROS17/2.8 with two PTH fragments harboring distinct activating domains: PTH-(1-34) and PTH-(28-48). The PTH response of genes expressed immediate early in the cell cycle or in the osteoblast developmental sequence was investigated. In addition, subtractive cloning was used to identify genes in ROS17/2.8 cells that are activated by the two PTH domains. PTH-(1-34) immediately increased the transcript levels of c-fos and c-jun at a considerably higher rate than PTH-(28-48). A significant immediate PTH effect on osteoblastic marker genes could not be detected, with the exception of elevated ornithine decarboxylase transcript levels. However, continuous application of PTH-(1-34) increased transcript levels of the osteoblast-specific osteocalcin gene and reduced those of other osteoblastic marker genes including alkaline phosphatase and the PTH/PTH related peptide receptor. By subtractive cloning, nine cDNAs were isolated corresponding to mRNAs directly up-regulated by PTH-(1-34) or PTH-(28-48). Among these were a cyclic phosphodiesterase, a (cytosine 5)-methyltransferase, an 80 kDa protein kinase C substrate, junB, and a novel GC-binding protein. Three cDNAs are unknown at present. Interestingly, in all cases, the efficiency of gene activation by PTH-(28-48) was substantially lower in comparison with PTH-(1-34). PTH-mediated protein kinase C signaling in ROS17/2.8 cells may therefore constitute a minor pathway in comparison with the dominant cAMP/protein kinase A cascade. PMID- 8702989 TI - Functional analysis of Shigella VirG domains essential for interaction with vinculin and actin-based motility. AB - The VirG (IcsA) protein of Shigella is required for recruitment of host actin filament (F-actin) by intracellularly motile bacteria. An N-terminal 80-kDa VirG portion (alpha-domain) is exposed on the bacterial surface, while the following C terminal 37-kDa portion (beta-core) is embedded in the outer membrane. Here, we report that the surface exposed alpha-domain of VirG possesses two distinct functional domains; one is the N-terminal two-thirds portion of the alpha-domain which is required for eliciting F-actin assembly on the bacteria in infected cells, and the other one is the rest of the C-terminal portion of the VirG alpha domain, which is essential for the asymmetric distribution of VirG on the bacterial surface. Furthermore, we found that vinculin, an actin-binding cytoskeletal protein, accumulates on the surface of bacteria expressing VirG in infected cells, and that the distribution of vinculin coincided with the distribution of VirG and assembled F-actin. The vinculin accumulation depended on the expression of the alpha-domain VirG portion required for F-actin assembly, but the recruitment of vinculin on Shigella appeared prior to the appearance of F actin in the infected cells. Analysis of proteins interacting with VirG using Xenopus laevis eggs extracts revealed that vinculin was a protein that bound to the alpha-domain portion. This was further confirmed using purified chicken gizzard vinculin, in that the 95-kDa vinculin head part, but not the 30-kDa tail part, directly bound to the alpha-domain portion. These results suggest a possible role for vinculin in recruitment of F-actin to the VirG moiety exposed on Shigella in infected mammalian cells. PMID- 8702990 TI - Antileukoprotease inhibits stratum corneum chymotryptic enzyme. Evidence for a regulative function in desquamation. AB - The stratum corneum chymotryptic enzyme (SCCE) has been previously purified from human stratum corneum and resembles a chymotryptic serine endopeptidase involved in physiological detachment of corneocytes from human stratum corneum. From human stratum corneum two inhibitory activities of SCCE could be extracted. These were due to serine protease inhibitors already known to be present in human epidermis, antileukoprotease (secretory leukocyte protease inhibitor) and elafin (skin derived antileukoprotease). The Inhibition of SCCE by antileukoprotease shows a hyperbolic, mixed type inhibition with an equilibrium dissociation constant of 63 n. Antileukoprotease also inhibits detachment of corneocytes from human plantar callus in vitro almost completely (>96%). In addition, elafin was shown to be a weak inhibitor for SCCE activity, and elafin significantly reduces the shedding of corneocytes. Thus, antileukoprotease, which is known to be produced by human keratinocytes, is likely to be the major physiological inhibitor of SCCE in the epidermis. It seems to be involved in the regulation of desquamation under physiological and pathophysiological conditions. PMID- 8702991 TI - dNTP binding to HIV-1 reverse transcriptase and mammalian DNA polymerase beta as revealed by affinity labeling with a photoreactive dNTP analog. AB - The dNTP binding pocket of human immunodeficiency virus type 1 reverse transcriptase (RT) and DNA polymerase beta (beta-pol) were labeled using a photoreactive analog of dCTP, exo-N-[beta-(p-azidotetrafluorobenzamido)-ethyl] deoxycytidine-5'- triphosphate (FABdCTP). Two approaches of photolabeling were utilized. In one approach, photoreactive FABdCTP and radiolabeled primer-template were UV-irradiated in the presence of each enzyme and resulted in polymerase radiolabeling. In an alternate approach, FABdCTP was first UV-cross-linked to enzyme; subsequently, radiolabeled primer-template was added, and the enzyme linked dCTP analog was incorporated onto the 3'-end of the radiolabeled primer. The results showed strong labeling of the p66 subunit of RT, with only minor labeling of p51. No difference in the intensity of cross-linking was observed with either approach. FABdCTP cross-linking was increased in the presence of a dideoxyterminated primer-template with RT, but not with beta-pol, suggesting a significant influence of prior primer-template binding on dNTP binding for RT. Mutagenesis of beta-pol residues observed to interact with the incoming dNTP in the crystal structure of the ternary complex resulted in labeling consistent with kinetic characterization of these mutants and indicated specific labeling of the dNTP binding pocket. PMID- 8702992 TI - Ordering the cell death pathway. Differential effects of BCL2, an interleukin-1 converting enzyme family protease inhibitor, and other survival agents on JNK activation in serum/nerve growth factor-deprived PC12 cells. AB - Previous studies indicate that activation of c-Jun kinase (JNK) is necessary for apoptosis of trophic factor-deprived PC12 cells and that death in this system is suppressed by multiple agents, including BCL2, inhibitors of the interleukin-1 converting enzyme (ICE) family of proteases, blockers of transcription, and a variety of small molecules with differing modes of action. Here, we determine the order in which these agents block apoptosis relative to JNK activation. Overexpression of BCL2 promotes PC12 cell survival and blocks JNK activation caused by trophic factor withdrawal. Similarly, the survival-promoting agents aurintricarboxylic acid, N-acetylcysteine, the nitric oxide generator diethylenetriamine nitric oxide, 8-bromo-cGMP, and 8-(4-chlorophenylthio)-cAMP act upstream to inhibit JNK activation. In contrast, zVAD-fluoromethylketone (a permeant ICE family inhibitor), actinomycin D, and the G1/S cell cycle inhibitor deferoxamine, all promote survival after trophic factor withdrawal, but do not affect JNK activation. These findings are consistent with the presence of an ordered cell death pathway triggered by trophic factor deprivation in which 1) BCL2 and a number of survival-promoting agents act upstream of JNK, 2) ICE family protease actions, regulated genes required for cell death, and certain cell cycle blockers lie either downstream of JNK or on independent pathways required for apoptotic death. PMID- 8702993 TI - Structure-function relationship of lipoprotein lipase-mediated enhancement of very low density lipoprotein binding and catabolism by the low density lipoprotein receptor. Functional importance of a properly folded surface loop covering the catalytic center. AB - We examined the structure-function relationship of human lipoprotein lipase (hLPL) in its ability to enhance the binding and catabolism of very low density lipoproteins (VLDL) in COS cells. Untransfected COS cells did not bind to or catabolize normal VLDL. Expression of wild-type hLPL by transient transfection enhanced binding, uptake, and degradation of the VLDL (a property of LPL that we call bridge function). Heparin pretreatment and a monoclonal antibody ID7 that blocks LDL receptor-binding domain of apoE both inhibited binding, and apoE2/E2 VLDL from a Type III hyperlipidemic subject did not bind. However, LDL did not reduce 125I-VLDL binding to the hLPL-expressing cells, whereas rabbit beta-VLDL was an effective competitor. By contrast, LDL reduced uptake and degradation of 125I-VLDL to the same extent as excess unlabeled VLDL or beta-VLDL. These data suggest that binding occurs by direct interaction of VLDL with LPL but the subsequent catabolism of the VLDL is mediated by the LDL receptor. Mutant hLPLs that were catalytically inactive, S132A, S132D, as well as the partially active mutant, S251T, and S172G, gave normal enhancement of VLDL binding and catabolism, whereas the partially active mutant S172D had markedly impaired capacity for the process; thus, there is no correlation between bridge function and lipolytic activity. A naturally occurring genetic variant hLPL, S447-->Ter, has normal bridge function. The catalytic center of LPL is covered by a 21-amino acid loop that must be repositioned before a lipid substrate can gain access to the active site for catalysis. We studied three hLPL loop mutants (LPL-cH, an enzymatically active mutant with the loop replaced by a hepatic lipase loop; LPL-cP, an enzymatically inactive mutant with the loop replaced by a pancreatic lipase loop; and C216S/C239S, an enzymatically inactive mutant with the pair of Cys residues delimiting the loop substituted by Ser residues) and a control double Cys mutant, C418S/C438S. Two of the loop mutants (LPL-cH and LPL-cP) and the control double Cys mutant C418S/C438S gave normal enhancement of VLDL binding and catabolism, whereas the third loop mutant, C216S/C239S, was completely inactive. We conclude that although catalytic activity and the actual primary sequence of the loop of LPL are relatively unimportant (wild-type LPL loop and pancreatic lipase loops have little sequence similarity), the intact folding of the loop, flanked by disulfide bonds, must be maintained for LPL to express its bridge function. PMID- 8702994 TI - Beta-amyloid precursor protein. Location of transmembrane domain and specificity of gamma-secretase cleavage. AB - The formation of beta-amyloid by processing of its precursor protein is a characteristic of Alzheimer's disease. Two proteolytic cleavages produce the amino and carboxyl termini of beta-amyloid, with the latter cleavage site located within the transmembrane domain. Using DNA mutagenesis, we investigated the membrane position and sequence requirements for carboxyl-terminal processing of the beta-amyloid domain. Substitution of negatively charged residues across positions 40-46 of the beta-amyloid domain precluded both beta-amyloid formation and precursor maturation associated with secretory protein transport. In contrast, identical substitutions from positions 48-50 had no adverse effects. Since charged residues typically prevent protein membrane insertion, these data define the membrane boundary to position 46/47, a location allowing greater access to carboxyl-terminal processing of beta-amyloid, possibly without membrane destruction. Deletions within the carboxyl-terminal domain, including 4 residues spanning positions 39-42 of beta-amyloid, resulted in formation of the beta amyloid peptide. Substituting residues 38-47 or 39-56 of the beta-amyloid domain in the precursor with a transmembrane sequence from another protein yielded a approximately 4-kDa beta-amyloid peptide, reflecting a loose residue specificity for carboxyl-terminal processing to beta-amyloid. PMID- 8702995 TI - Akt, a pleckstrin homology domain containing kinase, is activated primarily by phosphorylation. AB - Akt is a serine/threonine kinase that is stimulated by receptor tyrosine kinases and contains a pleckstrin homology domain. One model proposed to explain this activation suggests that receptor tyrosine kinases stimulate a phosphatidylinositol 3-kinase whose lipid products directly activate Akt kinase by interacting with its pleckstrin homology domain. In the present study, we show, in three cell types, that Akt does not require its pleckstrin homology domain to respond to either insulin or platelet-derived growth factor. Moreover, attachment of the src myristoylation signal to target Akt, without its pleckstrin homology domain, to the membrane constitutively activates Akt by causing an increase in its basal level of phosphorylation. This constitutively active form of Akt can also activate p70(S6K), indicating that the pleckstrin homology domain is not necessary for downstream interactions. Fusion of the inter src homology 2 domain from the p85 regulatory subunit of the phosphatidylinositol 3-kinase to Akt also constitutively activated Akt and induced an association with the lipid kinase. Phosphorylation of this fusion protein still critically contributes toward its increased activity. The sum of these results indicates that the primary mechanism of Akt activation is via protein phosphorylation. PMID- 8702996 TI - Structural topology of transmembrane helix 10 in the lactose permease of Escherichia coli. AB - In the lactose permease of Escherichia coli, transmembrane helix 10 has been shown to be functionally important. The structure of this helix has been examined in greater detail in this study. A total of 46 substitution and 8 insertional mutants were constructed and analyzed along the entire length of transmembrane helix 10. The results identified amino acids that are tolerant of substitutions by a variety of amino acids. Since a number of these amino acids (Thr-320, Val 331, Phe-325, and Ile-317) are clustered in one region in a helical wheel projection of transmembrane helix 10, it seems likely that this face of helix 10 is interacting with the membrane. The channel lining domain is thought to consist of the helical face containing Glu-325, Leu-318, Leu-329, His-322, Val-315, Cys 333, Val-326, and Lys-319 based on the results here and from earlier findings. Deleterious mutations along this face tended to greatly increase the Km value for lactose transport with only minor effects on the Vmax. Analysis of insertional mutants revealed that perturbation of the spatial relationship between amino acids at the periplasmic edge is less deleterious than perturbation in the center of the helix or the cytoplasmic edge. Using all of the above information, a detailed structural topology of transmembrane helix 10 is proposed. PMID- 8702997 TI - Phosphorylation of the inositol 1,4,5-trisphosphate receptor. Cyclic GMP dependent protein kinase mediates cAMP and cGMP dependent phosphorylation in the intact rat aorta. AB - The effects of cyclic GMP (cGMP) and activation of cGMP-dependent protein kinase (PKG) on the phosphorylation of the inositol 1,4, 5-trisphosphate (IP3) receptor were examined in intact rat aorta using the technique of back phosphorylation. Aorta treated with the nitric oxide donors, S-nitroso-N-acetylpenicillamine and sodium nitroprusside, or the selective PKG activator, 8-(4-para-chlorophenylthio) cGMP (8-CPT-cGMP), demonstrated increased IP3 receptor phosphorylation in situ, which was both time- and concentration-dependent with a stoichiometry of 0.5 mol of phosphate/mol of receptor above control. Treatment of aorta with the adenyl cyclase activator, forskolin, also demonstrated increased phosphorylation of the IP3 receptor on the PKG site, although the selective cAMP-dependent protein kinase activator, 8-(4-para-chlorophenylthio)-cAMP (8-CPT-cAMP), did not increase the phosphorylation of the IP3 receptor. Moreover, the PKG selective inhibitor, KT 5823, inhibited both sodium nitroprusside and forskolin-induced IP3 receptor phosphorylation more potently than the selective cAMP-dependent protein kinase inhibitor, KT 5720, suggesting that PKG mediates the increase in IP3 receptor phosphorylation by both cyclic nucleotides in intact aorta. These results provide further support for the notion that PKG is activated by both cAMP and cGMP in intact vascular smooth muscle and that PKG performs a critical role in cyclic nucleotide-dependent relaxation of blood vessels. PMID- 8702998 TI - Formation of c-Cbl.phosphatidylinositol 3-kinase complexes on lymphocyte membranes by a p56lck-independent mechanism. AB - The proto-oncogene c-Cbl was originally identified as a cellular homologue of the transforming protein expressed by the murine Cas NS-1 retrovirus. The full-length c-Cbl protein is a predominantly cytoplasmic protein, abundant in lymphoid cells, and potentially involved in signal transduction in several cell types. The specific signal transduction pathways in which c-Cbl participates, and its precise role in these pathways, are unclear. Previous studies from our laboratory have shown that c-Cbl is the predominant tyrosine-phosphorylated protein bound to the p85 subunit of phosphatidylinositol (PI) 3-kinase on T lymphocyte and B lymphocyte activation. To further understand the properties of c-Cbl and the significance of its interactions with PI 3-kinase, we have further studied the cellular biological and biochemical responses of c-Cbl to stimulation in lymphoid cells. We show that stimulation induces the association of a highly tyrosine phosphorylated pool of c-Cbl with lymphocyte membranes and with a detergent insoluble particulate fraction. Immunoprecipitation of c-Cbl from subcellular fractions reveals that p85 is predominantly associated with the c-Cbl pool recovered from the membrane fraction, despite the fact that this pool represents a small amount of total cellular c-Cbl. The formation of c-Cbl.PI 3-kinase complexes on lymphocyte membranes did not depend on the catalytic activity of PI 3-kinase since it was unaltered by the treatment of cells with wortmannin prior to stimulation. Interestingly, c-Cbl tyrosine phosphorylation and the formation of c-Cbl.PI 3-kinase complexes were also observed in a mutant Jurkat cell line, JCaM1.6, lacking p56(lck) expression. Because p56(lck) is critical for mitogenic signal transduction in response to T cell receptor activation, our results suggest that the activation of c-Cbl and the formation of c-Cbl.PI 3-kinase complexes occur upstream or independently of mitogenic signal transduction pathways in T cells. PMID- 8702999 TI - Characterization of a vacuolar protease in Neurospora crassa and the use of gene RIPing to generate protease-deficient strains. AB - We have isolated a gene from Neurospora crassa that appears to encode a pepstatin sensitive protease found both in membranes and in soluble contents of vacuoles. The gene contains two introns and encodes a 396-residue protein with a molecular mass of 42,900 Da. Because of the similarity of the protein to proteinase A in Saccharomyces cerevisiae the gene has been named pep-4. Strains with mutations in the pep-4 gene were generated in vivo by the gene RIPing procedure described by Selker and Garrett (Selker, E. U., and Garrett, P. W. (1988) Proc. Natl. Acad. Sci. U. S. A. 85, 6870-6874). The mutant strains were deficient in pepstatin sensitive protease activity and did not appear to produce a major 42-kDa polypeptide in the vacuole. The mutant strains grew at the same rate as the wild type and had no other observable phenotype. When compared with inactivation of the PEP4 gene of S. cerevisiae, inactivation of the pep-4 gene in N. crassa produced a phenotype that was different in several ways. In N. crassa the mutant strains did not exhibit reduced sporulation or reduced viability after nitrogen starvation, and they had elevated levels of proteinase B and carboxypeptidase activities. The pep-4 gene appears to encode the N. crassa, homolog of proteinase A, but the maturation of vacuolar hydrolases appeared to be less dependent on this protease than has been observed in S. cerevisiae. PMID- 8703000 TI - Thyroid hormone activation of transcription is potentiated by activators of cAMP dependent protein kinase. AB - We characterized the cross-talk between activators of protein kinase A (PKA) and thyroid hormone (T3) in T3 receptor (TR)-mediated transcription. U937 cells were cotransfected with a plasmid expressing the TR and a reporter plasmid containing a T3 response element (TRE) oriented either as a direct repeat or as a palindrome upstream of the thymidine kinase promoter linked to the chloramphenicol acetyltransferase gene. T3 activated transcription by 10-fold. T3 response was potentiated 2.5-3-fold by activators of PKA, but an activator of protein kinase C or of guanylate kinase was ineffective. In the absence of T3, activators of PKA had no effect on transcription. TR heterodimerization with the retinoid X receptor may facilitate T3/PKA cross-talk because coexpression of the retinoid X receptor potentiated cross-talk. Synergy was not observed in JEG-3, F9, CV-1, HeLa, L929, and HTC cells, indicating that it may require cell-specific factors. Synergy required the DNA- and ligand-binding domains, but not the amino-terminal domain, indicating that T3- and TRE-induced conformational changes on the TR are essential for cross-talk. PKA phosphorylated the TR in vitro, suggesting that, like other nuclear receptors, the TR is a target for PKA. These results imply that PKA cross-talks with T3 at the level of the TRE-bound TR, enhancing its transcriptional activity in a cell-specific manner. PMID- 8703001 TI - The 1.5-A resolution crystal structure of bacterial luciferase in low salt conditions. AB - Bacterial luciferase is a flavin monooxygenase that catalyzes the oxidation of a long-chain aldehyde and releases energy in the form of visible light. A new crystal form of luciferase cloned from Vibrio harveyi has been grown under low salt concentrations, which diffract x-rays beyond 1.5-A resolution. The x-ray structure of bacterial luciferase has been refined to a conventional R-factor of 18.2% for all recorded synchrotron data between 30.0 and 1.50-A resolution. Bacterial luciferase is an alpha-beta heterodimer, and the individual subunits fold into a single domain (beta/alpha)8 barrel. The high resolution structure reveals a non-prolyl cis peptide bond that forms between Ala74 and Ala75 in the alpha subunit near the putative active site. This cis peptide bond may have functional significance for creating a cavity at the active site. Bacterial luciferase employs reduced flavin as a substrate rather than a cofactor. The structure presented was determined in the absence of substrates. A comparison of the structural similarities between luciferase and a nonfluorescent flavoprotein, which is expressed in the lux operon of one genus of bioluminescent bacteria, suggests that the two proteins originated from a common ancestor. However, the flavin binding sites of the nonfluorescent protein are likely not representative of the flavin binding site on luciferase. The structure presented here will furnish a detailed molecular model for all bacterial luciferases. PMID- 8703002 TI - Long-acting growth hormones produced by conjugation with polyethylene glycol. AB - Derivatives of human growth hormone (hGH) of increasing size were produced by reaction with the N-hydroxysuccinimide ester of polyethylene glycol-5000 (PEG5000), a 5-kDa reagent that selectively conjugates to primary amines. By adjusting the reaction conditions and purification procedure, it was possible to isolate hGH derivatives containing up to seven PEG moieties that altered the Stokes radius and thereby the effective molecular masses of the unmodified hormone from 22 to 300 kDa. Fortunately, the most reactive amines were ones that did not lie in either of the two sites important for receptor binding. Nonetheless, increasing the level of PEG modification linearly reduced the affinity of hGH for its receptor and increased the EC50 in a cell-based assay up to 1500-fold. Most of the reduction in affinity was the result of slowing the association rate for the receptor. The clearance rate of hGH in rats was inversely proportional to effective molecular weight and closely fit a filtration model. We have tested the potency of these analogs by injecting them daily or every 6 days into hypophysectomized rats and determining the effects on body and organ growth. The efficacy of these analogs was optimal for hGH conjugated with 5 eq of PEG5000, and the potency was increased by about 10-fold compared with unmodified hGH. Such PEG-hGH derivatives show promise as long-acting alternatives to daily injections of hGH. More generally these studies show that improving hormone clearance properties, even at the expense of reducing receptor binding affinity, can lead to dramatic increases in hormone efficacy. PMID- 8703003 TI - Ligand binding to integrin alphaIIbbeta3 is dependent on a MIDAS-like domain in the beta3 subunit. AB - Substitution of beta3 residue Asp119, Ser121, or Ser123 results in a loss of the ligand binding function of integrin alphaIIbbeta3. Homologous residues in other integrin beta subunits are similarly critical for ligand binding function. This DXSXS motif is also present in the I domain of certain integrin alpha subunits, where it constitutes a portion of the unique metal ion-dependent adhesion site (MIDAS). In this report, we have utilized the crystal structure of the recombinant alphaM I domain to produce a three-dimensional model of the homologous region in the integrin beta3 subunit. We performed mutagenesis of candidate amino acid residues predicted from this model to be involved in cation coordination and ligand binding. We report the identification of Asp217 and Glu220 as residues essential for the ligand binding function of alphaIIbbeta3. Alanine substitution of these residues did not affect receptor expression but abolished the binding of activation-dependent (PAC1) and -independent (OPG2) ligand mimetic antibodies. In our proposed model, beta3 Asp217 is analogous to a metal-coordinating residue in the alphaM MIDAS domain, while Glu220 does not correspond to a functional MIDAS domain residue. Substitution of the highly conserved beta3 residue Thr197 corresponding to a critical MIDAS metal coordinating Thr residue did not affect ligand binding function, suggesting that this region of beta3 adopts a structure that is very similar to but not identical to that of the MIDAS domain. These data support a functional linkage between these two sequences and further define a common feature of ligand binding to integrins. PMID- 8703004 TI - Intracellular third loop domain of angiotensin II type-2 receptor. Role in mediating signal transduction and cellular function. AB - The present study tests the hypothesis that the unique intracellular third loop domain of angiotensin II type-2 (AT2) receptor is essential for the subsequent intracellular signaling and plays an important role in mediating receptor function. Synthetic intracellular third loop peptide of the AT2 receptor (AT2 3LP, 22 amino acids) and control peptide consisting of the same amino acid composition in random sequence were delivered into adult rat aortic vascular smooth muscle cells by cationic liposome-mediated transfection. Successful intracellular peptide delivery was confirmed by microscopic localization of the fluorescein-labeled AT2-3LP within the cells and also by co-immunoprecipitation of the 125I-labeled 3LP complexed with Gi protein using anti-Gialpha antibody. The AT2-3LP-transfected cells showed reduction of serum-stimulated DNA synthesis and cell proliferation as well as a decrease in mitogen-activated protein kinase activity, simulating the effects of AT2 receptor stimulation. The antagonistic effect of the AT2-3LP on mitogen-activated protein kinase activity and DNA synthesis were reversed by pertussis toxin and sodium orthovanadate. Thus, our data suggest that the intracellular third loop domain of the AT2 receptor is closely linked with the cellular signaling pathways of vascular smooth muscle cells in which Gi and protein-phosphotyrosine phosphatase are involved, resulting in the alteration of mitogen-activated protein kinase activity and in growth inhibition. PMID- 8703005 TI - Constitutive expression of the gene for the cell-specific p48 DNA-binding subunit of pancreas transcription factor 1 in cultured cells is under control of binding sites for transcription factors Sp1 and alphaCbf. AB - We have cloned and characterized the rat gene that encodes the p48 DNA-binding subunit of pancreas transcription factor 1 (Ptf1), a cell-specific basic region helix-loop-helix (bHLH) protein. The ptf1-p48 gene measures 1.8 kilobases in size and occurs as a single copy in the haploid genome. Run-on transcription assays suggest that this gene is subject to transcriptional control since no activity of its promoter is detected in nonproducing cells. The gene specifies two mRNAs that encode the same protein and originate from transcription initiation at alternative sites. Expression analysis of hybrid genes bearing deletions of the gene's 5'-flanking region fused to a reporter gene defines a promoter region within the gene-proximal 260 base pairs of DNA. The cis-acting elements that control promoter activity include binding sites for transcription factors Sp1 and alphaCbf, a 60-kDa CCAAT box-binding protein. The gene promoter, however, functions not only in exocrine pancreatic cells but also in cells of other origin. No cell-specific transcriptional control element was detected in as much as 10 kilobases of 5'-flanking region. We discuss models of how the cell-specific expression of the endogenous ptf1-p48 gene might be established during development of the animal. PMID- 8703006 TI - Mechanisms of desensitization and resensitization of proteinase-activated receptor-2. AB - Proteinase-activated receptor-2 (PAR-2) is a G-protein-coupled receptor that is expressed by intestinal epithelial cells, which are episodically exposed to pancreatic trypsin in the intestinal lumen. Trypsin cleaves PAR-2 to expose a tethered ligand, which irreversibly activates the receptor. Thus, PAR-2 may desensitize and resensitize by novel mechanisms. We examined these mechanisms in kidney epithelial cells, stably expressing human PAR-2, and intestinal epithelial cells, which naturally express PAR-2. Trypsin stimulated a prompt increase in [Ca2+]i, due to mobilization of intracellular Ca2+, followed by a sustained plateau, due to influx of extracellular Ca2+. Repeated application of trypsin caused marked desensitization of this response, which is due in part to (a) irreversible cleavage of the receptor by trypsin and (b) protein kinase C mediated termination of signaling. Trypsin exposure resulted in internalization of PAR-2 into early endosomes and then lysosomes; but endocytosis was not the mechanism of rapid desensitization. Thus, activated PAR-2 is endocytosed and degraded. The Ca2+ response to trypsin resensitized by 60-90 min. Brefeldin A, which disrupted Golgi stores of PAR-2, and cycloheximide, which inhibited protein synthesis, markedly attenuated resensitization. Thus, PAR-2-mediated Ca2+ mobilization desensitizes by irreversible receptor cleavage, protein kinase C mediated termination of signaling, and PAR-2 targeting to lysosomes. It resensitizes by mobilization of large Golgi stores and synthesis of new receptors. PMID- 8703007 TI - Gene for aspartate racemase from the sulfur-dependent hyperthermophilic archaeum, Desulfurococcus strain SY. AB - Amino acid racemases are ubiquitous throughout eubacteria. However, no amino acid racemases have yet been found in eukaryotes and archaea. We cloned a gene highly homologous to that for the aspartate racemase from the sulfur-dependent hyperthermophilic archaeum, Desulfurococcus strain SY. The product of the gene showed 35.2% amino acid sequence identity with the aspartate racemase of Streptococcus thermophilus IAM10064, and was also homologous to glutamate racemases around the putative catalytic cysteine residues. The encoded protein was expressed in Escherichia coli. The recombinant protein had amino acid racemizing activity, which was highly specific for aspartate and increased with temperature from 37 degrees C to 90 degrees C. Therefore, this was identified as the first hyperthermophilic archaeal amino acid racemase. A little aspartate racemizing activity was also detected in the crude extract of Desulfurococcus strain SY. The function of this aspartate racemase might be the uptake of aspartate formed at high temperature or the production of -aspartate as a cell component. The fact that the amino acid racemases are distributed among both eubacteria and archaea suggests that endogenous -amino acids in mammals are also synthesized by amino acid racemases. PMID- 8703008 TI - Repression of adipogenesis by adenylyl cyclase stimulatory G-protein alpha subunit is expressed within region 146-220. AB - The heterotrimeric G-protein alpha subunit stimulatory with respect to adenylyl cyclase (Gsalpha) represses adipogenesis of 3T3-L1 mouse embryonic fibroblasts. Derepression occurs in response to inducers, to oligodeoxynucleotides antisense to Gsalpha, and to overexpression of heterotrimeric G-protein alpha subunit 2, inhibitory with respect to adenylyl cyclase (Gialpha2). Constitutive expression of Gsalpha blocks adipogenesis and was exploited as an assay, in which chimeras of Gialpha2 and Gsalpha were expressed stably in 3T3-L1 cells to define the region controlling adipogenesis. N-terminal analysis revealed region 146-220 of Gsalpha as a repressor of adipogenesis; substitution of Gialpha2 abolished the ability of the chimera to repress adipogenesis in response to inducers. Expression of a chimera in which the 146-235 region of Gsalpha was embedded in Gialpha2 fully repressed adipogenesis in response to the inducers. C-terminal analysis revealed no loss of function for truncated Gsalpha, lacking the terminal 38 residues. The repressor domain for adipogenesis maps to a region that includes switch domains I and II and is spatially distinct from the regions mapped for control of adenylyl cyclase. PMID- 8703009 TI - Physical interaction of mammalian CDC37 with CDK4. AB - CDC37 was originally identified as a Start gene in budding yeast and has been shown to be required for association of CDC28 with cyclins. The exact functional mechanism by which CDC37 promotes this association, however, remains unknown. CDK4 is a cyclin D-dependent kinase that controls progression through G1 of the mammalian cell cycle. We have detected a specific association of CDK4 with the molecular chaperon HSP90 and a 44-kDa protein that we identify as mammalian CDC37. A physical interaction between CDC37 and CDK4 suggests that CDC37 may regulate the mammalian cell cycle through a direct effect on CDK4. Association of CDK4 with both CDC37 and HSP90 may also imply a mechanistic link between the functions of CDC37 and HSP90. PMID- 8703010 TI - The GAP-43 gene is a direct downstream target of the basic helix-loop-helix transcription factors. AB - The GAP-43 promoter region contains seven E-boxes (E1 to E7) that are organized in two clusters, a distal cluster (E3 to E7) and a proximal cluster (E1 and E2). Deletion analysis and site-directed mutagenesis of the GAP-43 promoter region showed that only the most proximal E1 E-box significantly modulates GAP-43 promoter activity. This E-box is conserved between the rat and human GAP-43 promoter sequences in terms of flanking sequence, core sequence (CAGTTG), and position. We found that endogenous E-box-binding proteins present in neuronal N18 cells recognize the E1 E-box and activate the GAP-43 promoter. The transcriptional activity of the GAP-43 promoter was repressed not only by the negative regulator Id2 protein, but also by two class A basic helix-loop-helix proteins, E12 and ME1a. In vitro analyses showed that both ME1a and E12 bind to the E1 E-box as homodimers. By Northern analyses, we established an inverse correlation between the level of E12 and ME1a mRNAs and GAP-43 mRNA in various neuronal cell lines as well as in ME1a-overexpressing PC12 cells. Therefore, we have identified a cis-acting element, the E1 E-box, located in the GAP-43 promoter region that modulates either positively or negatively the expression of the GAP-43 gene depending on which E-box-binding proteins occupy this site. Together, these data indicate that basic helix-loop-helix transcription factors regulate the expression of the GAP-43 gene and that the class A ME1a and E12 proteins act as down-regulators of GAP-43 expression. PMID- 8703011 TI - Induction of ceramide glucosyltransferase activity in cultured human keratinocytes. Correlation with culture differentiation. AB - Ceramides are the major component of the extracellular lipids that comprise the epidermal permeability barrier. They are derived from glucosylceramides (GlcCer) upon their extrusion from lamellar granules into the extracellular space in the upper layers of the epidermis. To better understand the regulation of the unique pathway for ceramide production in epidermis, we have studied the activity of the enzyme responsible for GlcCer synthesis, ceramide glucosyltransferase (CerGlc transferase), during keratinocyte culture differentiation. Human keratinocyte cultures were expanded in low calcium keratinocyte growth medium (KGM) and then switched to either normal calcium KGM (nKGM) or "complete" Dulbecco's modified Eagle's medium/Ham's F-12 (3:1) supplemented with 10% fetal bovine serum (cDMEM). At 7 and 10 days after the medium switch, electron microscopy revealed that cDMEM cultures were more fully differentiated morphologically and contained numerous lamellar granules. The GlcCer/DNA content of cDMEM cultures increased to 6 times that of day 0 cultures and was nearly 4 times greater than that of nKGM cultures, whereas the total lipid/DNA content of cDMEM cultures increased to only 1.8 times that of day 0 cultures and was approximately 1.2 times that of nKGM cultures. CerGlc transferase activity/DNA increased 6 times in cDMEM cultures but <1.5 times in nKGM cultures. By contrast, beta-glucocerebrosidase activity, which is responsible for the conversion of GlcCer to ceramide, increased to a similar extent in both differentiating culture systems. Treatment of cultures with the reversible CerGlc transferase inhibitor, DL-threo-1-phenyl-2-(palmitoylamino)-3 morpholino-1-propanol, prevented the increase of GlcCer in cDMEM cultures, and blocked conversion of exogenously added ceramide to GlcCer. A low level of CerGlc transferase activity, relative to that in differentiated keratinocytes, was detected in cultures of other human cell types. These results indicate that CerGlc transferase activity is induced during epidermal differentiation and that regulation of this enzyme may be an important determinant of the specialized production and compartmentalization of epidermal sphingolipids. PMID- 8703012 TI - Cloning and characterization of a novel membrane-associated lymphocyte NAD:arginine ADP-ribosyltransferase. AB - Mono-ADP-ribosylation is a post-translational modification of proteins in which the ADP-ribose moiety of NAD is transferred to proteins and is responsible for the toxicity of some bacterial toxins (e.g. cholera toxin and pertussis toxin). NAD:arginine ADP-ribosyltransferases cloned from human and rabbit skeletal muscle and from mouse lymphoma (Yac-1) cells are glycosylphosphatidylinositol-anchored and have similar enzymatic and physical properties; transferases cloned from chicken heterophils and red cells have signal peptides and may be secreted. We report here the cloning and characterization of an ADP-ribosyltransferase (Yac 2), also from Yac-1 lymphoma cells, that differs in properties from the previously identified eukaryotic transferases. The nucleotide and deduced amino acid sequences of the Yac-1 and Yac-2 transferases are 58 and 33% identical, respectively. The Yac-2 protein is membrane-bound but, unlike the Yac-1 enzyme, appears not to be glycosylphosphatidylinositol-anchored. The Yac-1 and Yac-2 enzymes, expressed as glutathione S-transferase fusion proteins in Escherichia coli, were used to compare their ADP-ribosyltransferase and NAD glycohydrolase activities. Using agmatine as the ADP-ribose acceptor, the Yac-1 enzyme was predominantly an ADP-ribosyltransferase, whereas the transferase and NAD glycohydrolase activities of the recombinant Yac-2 protein were equivalent. The deduced amino acid sequence of the Yac-2 transferase contained consensus regions common to several bacterial toxin and mammalian transferases and NAD glycohydrolases, consistent with the hypothesis that there is a common mechanism of NAD binding and catalysis among ADP-ribosyltransferases. PMID- 8703013 TI - Biosynthesis and expression of polysialic acid on the neural cell adhesion molecule is predominantly directed by ST8Sia II/STX during in vitro neuronal differentiation. AB - We have reported recently that ST8Sia II/STX as well as ST8Sia IV/PST-1 is a neural cell adhesion molecule (NCAM)-specific polysialic acid (PSA) synthase (Kojima, N., Tachida, Y., Yoshida, Y., and Tsuji, S. (1996) J. Biol. Chem. 271, 19457-19463). To investigate which of two PSA synthase (ST8Sia II and IV) are involved in the biosynthesis of PSA associated with NCAM, the expressions of PSA, PSA synthase activity, and the genes of two PSA synthases during in vitro neuronal differentiation of mouse embryonal carcinoma P19 cells were determined. PSA was not expressed on undifferentiated cells (day 0) or cell aggregates (days 1-3) induced with retinoic acid. Expression of PSA began after cell aggregates had been dissociated and re-plated on a dish (day 4) and increased up to day 7. The expression of the mouse ST8Sia II gene was negligible in both undifferentiated and aggregated cells, it beginning at day 4, then dramatically increasing, and reaching the maximum level at days 6-7. On the other hand, transcription of the ST8Sia IV gene remained at a very low level throughout the entire period, a significant increase in its expression during differentiation not being observed. PSA synthase activity was not detected in undifferentiated or aggregated P19 cells, it increasing in parallel with ST8Sia II gene expression during differentiation. In addition, the cells at day 7 were stained with an anti mouse ST8Sia II antiserum. Similar up-regulation of the ST8Sia II gene were observed during the differentiation of rat MNS-8 cells, which were derived from E 12 rat neuroepithelium of the neural tube and shown to differentiate into neurons. These results indicate that ST8Sia II predominantly directs PSA expression during neuronal differentiation rather than ST8Sia IV. PMID- 8703014 TI - Regulation of angiotensin II-stimulated Ca2+ oscillations by Ca2+ influx mechanisms in adrenal glomerulosa cells. AB - In adrenal glomerulosa cells, angiotensin II (Ang II) evokes repetitive [Ca2+]i transients and increases Ca2+ influx through voltage-sensitive calcium channels (VSCCs) as well as the capacitative Ca2+ entry pathway. This study analyzed the relationships between these Ca2+ influx pathways and intracellular Ca2+ signaling in bovine glomerulosa cells, in which Ca2+ oscillation frequency was regulated by Ang II concentration over the range of 50-300 p. In the absence of external Ca2+, such oscillations were maintained for prolonged periods of time, but their frequency was significantly reduced (0.23 min-1 versus 0.38 min-1). Restoration of [Ca2+]o to 0.6 mM increased the frequency of Ca2+ oscillations in cells that showed narrow spikes of constant amplitude and caused a plateau response in cells with broad spikes of rapidly decreasing amplitude. In the presence of Ca2+, nifedipine reduced the frequency of the oscillatory Ca2+ response to 100 pM Ang II by 49%, and BAY K 8644 increased oscillation frequency by 86%, or caused plateau-type responses typical of higher Ang II concentrations. In contrast to their prominent actions on Ca2+ spiking frequency, dihydropyridines caused only minor changes in Ang II (100 pM)-induced inositol phosphate production. Dihydropyridines also had minimal effects on the nonoscillatory Ca2+ signals evoked by high Ang II concentrations (10 nM). These findings indicate that Ca2+ influx through VSCCs modulates the frequency of Ca2+ oscillations induced by low agonist concentrations by a mechanism that does not involve major changes in inositol trisphosphate formation. However, VSCCs make relatively little contribution to the nonoscillatory Ca2+ signals generated by high agonist concentrations, when Ca2+ influx occurs predominantly through the capacitative Ca2+ entry pathway. PMID- 8703015 TI - Adenovirus-mediated expression of aquaporin-5 in epithelial cells. AB - A recombinant adenovirus coding for rat aquaporin-5 was constructed and plaque purified. The recombinant adenovirus (AdrAQP5) mediated the expression of aquaporin-5 in rat and human salivary cell lines and in dog kidney cells in vitro as demonstrated by Northern blot and Western blot analyses, and by confocal microscopy after immunofluorescent labeling. In kidney cells, expression of the transgene was optimal if cells were infected at their basolateral surface, a phenomenon associated with the distribution of integrin receptors on these cells. The expressed aquaporin-5 protein was functionally active because viral-mediated gene transfer resulted in a significant increase in the osmotically directed net fluid secretion rate across monolayers of kidney cells. AdrAQP5 should provide an efficient and useful means to impart facilitated water permeability to cells lacking such a pathway. PMID- 8703016 TI - Identification of a mutation in a GATA binding site of the platelet glycoprotein Ibbeta promoter resulting in the Bernard-Soulier syndrome. AB - Bernard-Soulier Syndrome (BSS) is a rare congenital bleeding disorder due to absent or decreased expression of the glycoprotein Ib-IX-V (GpIb-IX-V) receptor complex on the platelet surface. To date, only mutations in GpIbalpha or GpIX have been reported in patients with BSS. GpIbbeta differs from the other proteins in this receptor in that the gene is more complex, and an alternative form is expressed in cells of non-megakaryocytic lineage, including endothelial cells. It appears that the megakaryocytic and endothelial cell mRNA species are transcribed from different start sites and have different proximal promoter regions. We have identified a patient with BSS who has a deletion on one chromosome 22, resulting in velocardiofacial syndrome. The GpIbbeta gene has been mapped to this deleted (22q11.2) region of chromosome 22. The patient has greatly reduced levels of GpIbbeta mRNA and no detectable platelet GpIbbeta protein, suggesting that his BSS results from a mutation in his remaining GpIbbeta allele. Sequence analysis revealed that the coding region of GpIbbeta is normal, but the 5'-upstream region contains a C to G transversion at base -133 from the transcription start site used in megakaryocytes. The mutation changes a GATA consensus binding site, disrupts GATA-1 binding to the mutated site, and decreases promoter activity by 84%. Thus, in this patient, Bernard-Soulier syndrome results from a deletion of one copy of GpIbbeta and a mutated GATA binding site in the promoter of the remaining allele, resulting in decreased promoter function and GpIbbeta gene transcription. PMID- 8703017 TI - The B56 family of protein phosphatase 2A (PP2A) regulatory subunits encodes differentiation-induced phosphoproteins that target PP2A to both nucleus and cytoplasm. AB - Protein phosphatase 2A is a heterotrimeric protein serine/threonine phosphatase consisting of a 36-kDa catalytic C subunit, a 65-kDa structural A subunit, and a variable regulatory B subunit. The B subunits determine the substrate specificity of the enzyme. There have been three families of cellular B subunits identified to date: B55, B56 (B'), and PR72/130. We have now cloned five genes encoding human B56 isoforms. Polypeptides encoded by all but one splice variant (B56gamma1) are phosphoproteins, as shown by mobility shift after treatment with alkaline phosphatase and metabolic labeling with [32P]phosphate. All labeled isoforms contain solely phosphoserine. Indirect immunofluorescence microscopy demonstrates distinct patterns of intracellular targeting by different B56 isoforms. Specifically, B56alpha, B56beta, and B56epsilon complexed with the protein phosphatase 2A A and C subunits localize to the cytoplasm, whereas B56delta, B56gamma1, and B56gamma3 are concentrated in the nucleus. Two isoforms (B56beta and B56delta) are highly expressed in adult brain; here we show that mRNA for these isoforms increases severalfold when neuroblastoma cell lines are induced to differentiate by retinoic acid treatment. These studies demonstrate an increasing diversity of regulatory mechanisms to control the activity of this key intracellular protein phosphatase and suggest distinct functions for isoforms targeted to different intracellular locations. PMID- 8703018 TI - Characterization and hormonal modulation of anticoagulant heparan sulfate proteoglycans synthesized by rat ovarian granulosa cells. AB - Anticoagulant heparan sulfate proteoglycans endow the vascular endothelium with antithrombotic properties, but their role outside the vascular bed is unknown. Granulosa cells form an avascular compartment in the ovarian follicle, in which a heparin-like activity has been described. At ovulation extravascular coagulation occurs around ovulatory follicles, and after expulsion of the oocyte, a fibrin clot forms in the antral cavity. Granulosa cells synthesize two major heparan sulfate proteoglycans, whose anticoagulant nature has not been investigated. The purpose of this study was to characterize anticoagulant heparan sulfate proteoglycans synthesized by rat ovarian granulosa cells. Affinity purified 35S labeled anticoagulant heparan sulfate glycosaminoglycans represent 6.5% of the total heparan sulfate synthesized, and they contain 13% 3-O-sulfated disaccharides that are markers of the antithrombin-binding site of heparin. The biological activity of granulosa cell heparan sulfate proteoglycans was demonstrated by their ability to bind antithrombin and to accelerate the formation of thrombin-antithrombin complexes. The impact of hormonal stimulation on granulosa cell anticoagulant heparan sulfate proteoglycans was studied using 125I-antithrombin binding assays. Follicle-stimulating hormone induced a redistribution of anticoagulant heparan sulfate proteoglycans from the granulosa cell layer to the culture medium, indicating that their distribution could be modulated according to the stage of follicular development. These results suggest that anticoagulant heparan sulfate might be critically located in the follicle to maintain fluidity around the oocyte until its expulsion at ovulation. PMID- 8703019 TI - Insulin induces tyrosine phosphorylation of JAK2 in insulin-sensitive tissues of the intact rat. AB - The Janus kinase family of protein tyrosine kinases constitutes a novel type of signal transduction pathway activated in response to a wide variety of polypeptide ligands and has four known members: JAK1, JAK2, JAK3, and Tyk2. In this study, we examined the ability of insulin to stimulate JAK2 tyrosine phosphorylation in insulin-sensitive tissues of the intact rat using immunoprecipitation and immunoblotting. The results demonstrate that after an infusion of insulin, JAK2 is rapidly tyrosine phosphorylated (and the kinase is activated) in the liver, adipose tissue, skeletal muscle, heart, and isolated adipocytes. The presence of phosphorylated JAK2 was detectable after an infusion of insulin that increased serum insulin to physiological postprandial levels (40 70 microunits/ml). Co-immunoprecipitation with anti-insulin receptor antibody, anti-JAK2 antibody, and anti-IRS-1 antibody showed that JAK2 interacts with the insulin receptor and IRS-1 to form stable complexes following stimulation by insulin. In two animal models of insulin resistance the regulation of JAK2 tyrosine phosphorylation after insulin infusion paralleled the phosphorylation of the insulin receptor and of IRS-1. In conclusion, our data indicate that after physiological stimulation by insulin in the intact animal, JAK2 associates with the insulin receptor and is tyrosine phosphorylated in insulin-sensitive tissues in a time- and dose-dependent fashion. PMID- 8703020 TI - Protein import into mammalian mitochondria. Characterization of the intermediates along the import pathway of the precursor into the matrix. AB - We have characterized several intermediates in the mitochondrial import stimulation factor (MSF)-dependent import into mammalian mitochondria of a matrix targeted precursor, preadrenodoxin (pAd). In the first step, pAd docks onto the 37-kDa protein of the outer membrane (OM37) as a complex with MSF (stage I intermediate). It is then transferred to the import pore of OM in the presence of ATP, but in the absence of Deltapsi across the inner membrane (IM), to form stage II intermediate. Depletion of matrix ATP in the presence of both extramitochondrial ATP and Deltapsi induces accumulation of stage III intermediate, which is a mixture of the precursor with different intramitochondrial localizations: the precursor whose presequence had crossed either OM (IIIa) or both OM and IM (IIIb), but with a bulk portion remaining exposed to the cytosol and the precursor whose presequence had crossed both membranes, but with a residual portion staying within the intermembrane space (IIIc). These intermediates are on the correct import pathway and are characteristic in their protease accessibility, salt extractability, and antibody accessibility, as well as in their energy requirement for the chase reaction. PMID- 8703021 TI - Transcription-positive cofactor 4 forms complexes with HSSB (RPA) on single stranded DNA and influences HSSB-dependent enzymatic synthesis of simian virus 40 DNA. AB - The replication of simian virus 40 (SV40) DNA in vitro requires a trimeric single stranded DNA (ssDNA)-binding protein called HSSB or RPA. HSSB supports the unwinding of DNA containing the SV40 origin in the presence of the viral-encoded T antigen and is required for the initiation of RNA primer synthesis as well as processive elongation of DNA catalyzed by the DNA polymerase delta holoenzyme. In this report we show that the transcription positive cofactor 4 (PC4), a ssDNA binding protein, forms complexes with HSSB on ssDNA and markedly affects the replication functions of HSSB. PC4 supports T antigen-catalyzed unwinding of SV40 origins in lieu of HSSB but inhibits both RNA primer synthesis and polymerase delta-catalyzed DNA chain elongation reactions. These inhibitory effects can be reversed by the addition of excess HSSB. Depending on the concentration of HSSB, PC4 is capable of either inhibiting or activating SV40 DNA replication measured in both mono- and dipolymerase systems. The possible role of PC4 in the initiation of DNA replication is discussed. PMID- 8703022 TI - Interaction of kinesin motor domains with alpha- and beta-tubulin subunits at a tau-independent binding site. Regulation by polyglutamylation. AB - Interaction of rat kinesin and Drosophila nonclaret disjunctional motor domains with tubulin was studied by a blot overlay assay. Either plus-end or minus-end directed motor domain binds at the same extent to both alpha- and beta-tubulin subunits, suggesting that kinesin binding is an intrinsic property of each tubulin subunit and that motor directionality cannot be related to a preferential interaction with a given tubulin subunit. Binding features of dimeric versus monomeric rat kinesin heads suggest that dimerization could drive conformational changes to enhance binding to tubulin. Competition experiments have indicated that kinesin interacts with tubulin at a Tau-independent binding site. Complementary experiments have shown that kinesin does not interact with the same efficiency with the different tubulin isoforms. Masking the polyglutamyl chains with a specific monoclonal antibody leads to a complete inhibition of kinesin binding. These results are consistent with a model in which polyglutamylation of tubulin regulates kinesin binding through progressive conformational changes of the whole carboxyl-terminal domain of tubulin as a function of the polyglutamyl chain length, thus modulating the affinity of tubulin for kinesin and Tau as well. These results indicate that microtubules, through tubulin polymorphism, do have the ability to control microtubule-associated protein binding. PMID- 8703023 TI - Characterization of the upstream sequence of the human CYP11A1 gene for cell type specific expression. AB - The CYP11A1 gene encodes the cholesterol side-chain cleavage enzyme P450scc, which catalyzes the synthesis of steroids from cholesterol. This gene is expressed only in steroidogenic organs such as the adrenal, gonad, placenta, and brain. We have characterized an upstream regulatory element of the human CYP11A1 gene, termed AdE, which contributed to its cell type-specific expression. The AdE sequence contains two protein binding regions, AdE1 and AdE2, which bind many proteins including NF1- and Sp1-like proteins as shown by electrophoretic mobility shift assay, footprinting, competition, antibody supershift, and mutagenesis of the binding sites. When cloned in front of the CYP11A1 promoter or the heterologous thymidine kinase promoter, AdE sequences enhanced expression of the reporter gene in steroidogenic cell lines of the adrenal, gonad, and placental origin but not in nonsteroidogenic cell lines such as COS-1 and Rat-1. The function of AdE1 and AdE2 was lower when present individually than together. The combined action of multiple transcription factors binding to the AdE sequence brings about the final activation of the CYP11A1 gene in a tissue-specific manner. PMID- 8703024 TI - Isolation and amino acid sequence of a new 22-kDa FKBP-like peptidyl-prolyl cis/trans-isomerase of Escherichia coli. Similarity to Mip-like proteins of pathogenic bacteria. AB - We identified a periplasmic peptidyl-prolyl cis/trans-isomerase (PPIase) of the (FK506-binding protein (FKBP) type in Escherichia coli (FK506 represents a natural peptidomacrolide containing an acylated pipecolic acid residue). After purification to homogeneity, its complete amino acid sequence was determined by a combination of Edman degradation and electrospray mass spectrometry of the authentic protein and peptides generated by proteolysis. The molecular mass calculated from the amino acid sequence of the protein was 22,085.53 Da, which corresponded perfectly with the value of 22,084 +/- 1.47 Da as determined by mass spectrometry. The corresponding gene was cloned and analyzed, and Southern blot experiments revealed the existence of similar genes in various Gram-negative bacteria. The amino acid sequence of the novel FKBP22 shows similarity to Mip (macrophage infectivity potentiator)-like proteins produced by a number of pathogenic bacteria. However, FKBP22 is inhibited more strongly by FK506 than are other Mip-homologues, as indicated by the Ki value of 25 nM. The subsite specificity regarding the P1 position of the substrate resembles that for Mip FKBP25 from Legionella pneumophila. The mature FKBP22 enzyme of 205 amino acids exists as a dimer in solution. PMID- 8703025 TI - Partial purification and immunohistochemical localization of ATP diphosphohydrolase from Schistosoma mansoni. Immunological cross-reactivities with potato apyrase and Toxoplasma gondii nucleoside triphosphate hydrolase. AB - ATP diphosphohydrolase from tegumental membranes of Schistosoma mansoni was solubilized with Triton X-100 plus deoxycholate and separated by preparative nondenaturing polyacrylamide gel electrophoresis. Two isoforms with ATP hydrolytic activity were identified and excised from nondenaturing gels. For each of the active bands, two protein bands (63 and 55 kDa) were detected with Coomassie Blue staining, following sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Western blots developed with polyclonal anti-potato apyrase antibody revealed a single protein of 63 kDa, either with samples excised from active bands or with total S. mansoni tegument. Anti-potato apyrase antibody immobilized on Sepharose-Protein A depleted over 95% of ATPase and ADPase activities from detergent-solubilized tegument. Confocal laser scanning microscopy showed anti-potato apyrase antibody on the outer surface of S. mansoni tegument. A different antibody against a fusion protein derived from recently cloned Toxoplasma gondii nucleoside triphosphate hydrolase (Bermudes, D., Peck, K. R., Afifi, M. A., Beckers, C. J. M., and Joiner, K. A. (1994) J. Biol. Chem. 269, 29252-29260) revealed the same 63-kDa band in Western blots of S. mansoni tegument. Since anti-potato apyrase antibodies exhibited cross-reactivity with S. mansoni ATP diphosphohydrolase, we decided to gain further information on the primary structure of potato apyrase by sequencing the protein. Three novel peptides were obtained: amino-terminal sequence and two internal sequences from tryptic fragments. Eight sequences recently deposited in the data bank, including that of T. gondii nucleoside triphosphate hydrolase, have considerable homologies to potato apyrase suggesting a new family of nucleoside triphosphatases which contains a conserved motif (I/V)(V/M/I)(I/L/F/C)DAGS(S/T) near the amino terminal. Antibody cross-reactivities in the present work suggest that conserved epitopes from S. mansoni ATP diphosphohydrolase are present in this family of nucleotide-splitting enzymes. PMID- 8703026 TI - Alternative splicing in the N-terminal extracellular domain of the pituitary adenylate cyclase-activating polypeptide (PACAP) receptor modulates receptor selectivity and relative potencies of PACAP-27 and PACAP-38 in phospholipase C activation. AB - Pituitary adenylate cyclase-activating polypeptide (PACAP)-27 and PACAP-38 are neuropeptides of the vasoactive intestinal peptide/secretin/glucagon family. We previously described alternative splicing of the region encoding the third intracellular loop of the PACAP receptor generating six isoforms with differential signal transduction properties (Spengler, D., Waeber, C., Pantaloni, C., Holsboer, F., Bockaert, J., Seeburg, P. H., and Journot, L. (1993) Nature 365, 170-175). In addition, we demonstrated that the potencies of the two forms of PACAP are similar for adenylate cyclase stimulation, whereas PACAP-38 is more potent than PACAP-27 in phospholipase C activation. In the present work, we document the existence of a new splice variant of the PACAP receptor that was characterized by a 21-amino-acid deletion in the N-terminal extracellular domain. We demonstrate that this domain modulates receptor selectivity with respect to PACAP-27 and -38 binding and controls the relative potencies of the two agonists in phospholipase C stimulation. PMID- 8703027 TI - Assembly and activation of the phagocyte NADPH oxidase. Specific interaction of the N-terminal Src homology 3 domain of p47phox with p22phox is required for activation of the NADPH oxidase. AB - The phagocyte NADPH oxidase is activated during phagocytosis to produce superoxide, a precursor of microbicidal oxidants. The activation involves assembly of membrane-integrated cytochrome b558 comprising gp91(phox) and p22(phox), two specialized cytosolic proteins (p47(phox) and p67(phox)), each containing two Src homology 3 (SH3) domains, and the small G protein Rac. In the present study, we show that the N-terminal SH3 domain of p47(phox) binds to the C terminal cytoplasmic tail of p22(phox) with high affinity (KD = 0.34 microM). The binding is specific to this domain among several SH3 domains including the C terminal one of p47(phox) and the two of p67(phox) and requires the Pro156 containing proline-rich sequence but not other putative SH3 domain-binding sites of p22(phox). Replacement of Trp193 by Arg in the N-terminal SH3 domain completely abrogates the association with p22(phox). A mutant p47(phox) with this substitution is incapable of supporting superoxide production under cell-free activation conditions. These findings provide direct evidence that the interaction between the N-terminal SH3 domain of p47(phox) and the proline-rich region of p22(phox) is essential for activation of the NADPH oxidase. PMID- 8703028 TI - Protein splicing involving the Saccharomyces cerevisiae VMA intein. The steps in the splicing pathway, side reactions leading to protein cleavage, and establishment of an in vitro splicing system. AB - Protein splicing involves the excision of an internal protein segment, the intein, from a precursor protein and the concomitant ligation of the flanking N- and C-terminal regions. It occurs in mesophilic bacteria, yeast, and thermophilic archaea. The ability to control protein splicing of a thermophilic intein by temperature and pH in a foreign protein context facilitated the study of the mechanism of protein splicing in thermophiles. On the other hand, no direct studies have been done on the mechanism of protein splicing in mesophiles. We examined the splicing of a chimeric protein containing the intein of the vacuolar ATPase subunit (VMA) of Saccharomyces cerevisiae that involves cysteines rather than serines at the reaction center. The steps in the splicing process were deduced by analyzing intermediates and side products that accumulated as a result of amino acid substitutions and were found to be analogous to those occurring in thermophiles. Moreover, appropriate amino acid replacements allowed us to develop the first mesophilic in vitro protein splicing system as well as strategies for modulating the rate of protein splicing and for converting the splicing reaction to an efficient protein cleavage reaction at either splice junction. PMID- 8703029 TI - Post-transcriptional stabilization by interleukin-1beta of interleukin-6 mRNA induced by c-kit ligand and interleukin-10 in mouse bone marrow-derived mast cells. AB - We demostrate that a specific combination of cytokines elicits high levels of interleukin (IL)-6 gene expression in mast cells and define the cellular mechanisms of the exogenous cytokine action. The addition of c-kit ligand (KL) and IL-10 to IL-3-derived mouse bone marrow mast cells (BMMC) elicited an approximately 2-fold increase in steady-state IL-6 mRNA levels that peaked after 0.5 h and was followed by the release of approximately 0.2 ng of IL-6/10(6) cells by 5-7 h. The addition of IL-1beta to KL + IL-10 elicited a prolonged approximately 12-fold increase in the level of IL-6 mRNA by 3-5 h and an approximately 50-fold increase in the level of IL-6 protein released by 7 h. As determined by nuclear run-on analysis, KL + IL-10 stimulated IL-6 gene transcription within 0.5 h, and the addition of IL-1beta did not increase transcription. Instead, IL-1beta slowed by approximately 8-fold the decay of IL-6 mRNA as compared to its decay in BMMC stimulated with KL + IL-10 alone. The exposure of BMMC to cycloheximide 0.5 h before the addition of the three exogenous cytokines inhibited by approximately 50% the level of IL-6 mRNA generated but did not inhibit the effects of KL + IL-10, indicating that IL-1beta induces the synthesis of a protein that stabilizes IL-6 mRNA. The stabilization of IL-6 mRNA was inhibited by the addition of actinomycin D at 0.5 but not 3 h after BMMC were stimulated with IL-1beta in combination with KL + IL-10, suggesting that once transcribed, the stabilizing protein is long-lived. The addition of cycloheximide to BMMC after stimulation with KL + IL-10 with or without IL-1beta increased the levels of steady-state IL-6 mRNA compared to levels in cells without drug, indicating that in addition to stimulating IL-6 transcription, KL + IL-10 induces a protein factor that destabilizes IL-6 mRNA. Thus, there exists a novel Fcepsilon receptor type I-independent mechanism by which a mast cell can provide substantial amounts of IL-6 protein in response to the synergistic action of KL and IL-10 to induce IL-6 gene transcription, and IL 1beta to stabilize otherwise short-lived IL-6 transcripts. PMID- 8703030 TI - Stat6 and Jak1 are common elements in platelet-derived growth factor and interleukin-4 signal transduction pathways in NIH 3T3 fibroblasts. AB - Both platelet-derived growth factor (PDGF) and interleukin-4 (IL-4) play major roles in cell proliferation, differentiation, chemotaxis, and other functional responses. Here, we demonstrate that Stat6, previously shown to be activated by only IL-4 and IL-3, becomes activated after PDGF stimulation of NIH 3T3 fibroblasts. PDGF BB, and to a lesser extent PDGF AA, rapidly induced DNA binding activity from NIH 3T3 cell lysates utilizing the immunoglobulin heavy chain germ line epsilon promoter (Iepsilon) that specifically binds to Stat6 in an electrophoretic mobility shift assay. DNA binding activity could be detected within 5 min and reached maximum levels at approximately 20 min in parental NIH 3T3 cells. An identical mobility shift and time course of PDGF-mediated Iepsilon binding activity was more pronounced in lysates of NIH 3T3 transfectants overexpressing human Stat6 (NIH 3T3-Stat6). The observed radiolabeled Iepsilon mobility shift was competed by unlabeled Iepsilon as well as by the beta-casein gene promoter but not by the interferon-alpha-stimulated response element or the interferon-gamma response region of the guanylate-binding protein gene. A Stat6 specific polyclonal antisera also supershifted the PDGF-induced Iepsilon mobility shift. After PDGF BB treatment, a 100-kDa tyrosine phosphorylated species was detected in anti-Stat6 immunoprecipitates. Cycloheximide had little effect on Stat6 tyrosine phosphorylation. In addition to Stat6, Stat5a, and Stat5b, PDGF BB also induced Jak1 tyrosine phosphorylation suggesting a potential pathway for Stat activation. Strikingly, the concurrent addition of IL-4 enhanced PDGF BB induced Iepsilon binding activity, Jak1 tyrosine phosphorylation, and [3H]thymidine incorporation. These results provide evidence that Stat6 and Jak1 are common elements in PDGF and IL-4 signaling pathways and suggest that IL-4 could play a role in potentiating certain known PDGF-induced biological responses. PMID- 8703031 TI - Genomic organization and promoter function of the human thyrotropin-releasing hormone receptor gene. AB - We isolated and characterized the gene for the human thyrotropin-releasing hormone receptor. The gene spanned more than 30 kilobases and contained three exons and two introns. Intron 1 exists in the 5'-untranslated region, and intron 2 is more than 25 kilobases in length which interrupts the coding region before the beginning of the putative sixth transmembrane domain. Exon 3 encodes the rest of the coding region and the entire 3'-untranslated region. The 3'-flanking region contains four potential polyadenylation signals, and 3'-rapid amplification of cDNA ends studies showed that only a signal at 2076 base pairs downstream of the stop codon was functional in the anterior pituitary. Primer extension and anchor-polymerase chain reaction studies indicated a transcriptional start site at 344 base pairs upstream of the translational start site. The promoter region does not contain either a TATA box or a CAAT box in the appropriate location. Transient transfection study revealed significant activity of the promoter in GH4C1 cells, and the region between -338 and -933 bp from the transcriptional start site worked as a negative regulator. Knowledge of the genomic organization and the promoter region of thyrotropin-releasing hormone (TRH) receptor gene will allow further studies of possible disorders of the TRH receptor, as well as facilitate elucidation of transcriptional control of the human TRH receptor gene. PMID- 8703032 TI - Regulation of the RNA polymerase I and III transcription systems in response to growth conditions. AB - To better understand the mechanisms that regulate stable RNA synthesis, we have analyzed the RNA polymerase I and III transcriptional activities of extracts isolated from cells propagated under a variety of conditions. Under balanced growth conditions the levels of both RNA polymerase I- and III-specific transcription increased proportionally with growth rate. Upon nutritional starvation, RNA polymerase I transcription rapidly declined, followed by 5 S rDNA and eventually tDNA transcription. Transcriptional activities in extracts were restored when the nongrowing cultures were resuspended in fresh medium, although growth did not resume. The differential expression of 5 S rDNA and tDNA genes in extracts prepared from cells subjected to partial starvation was traced to a 5 S rDNA-specific inhibitor and not to a defect in any RNA polymerase III transcription factor. Characterization of this inhibitor indicated that it was not 5 S rRNA. It was sensitive to phenol extraction and resistant to RNase, and its target did not appear to be transcription factor IIIA. Not all treatments that slowed or stopped growth down-regulated the stable RNA transcription apparatus. Cells that have been subjected to either energy starvation or cycloheximide treatment still retain the ability to synthesize stable RNA in vitro, suggesting the presence of alternative regulatory mechanisms. PMID- 8703033 TI - Poly(3-hydroxybutyrate) is associated with specific proteins in the cytoplasm and membranes of Escherichia coli. AB - Poly(3-hydroxybutyrate) (PHB) is well-known as a high molecular weight homopolymer of R-3-hydroxybutyrate which accumulates in storage granules within the cytosols of certain bacteria. Escherichia coli does not amass these granules; however, small amounts of low molecular weight PHB (<0.02% of dry weight) have been found in the plasma membranes in complexes with calcium polyphosphate; the complexes serve as voltage-activated calcium channels. Here we report that polyphosphate-complexed PHB is only a minor fraction of the polyester in E. coli. PHB comprises 0.36 to 0. 55% of the dry weight of log-phase cells, depending on culture medium, and this amount increases by 15 to 20% when the cells are made genetically competent. The PHB is widely distributed throughout the cell, wherein it is primarily associated with proteins. The identity of protein-associated PHB was established by antibody reaction, chemical assay, and 1H NMR spectroscopy. As expected, the physical and chemical properties of protein-associated PHB were found to be considerably different from those of the bulk polymer or granule PHB, e.g. protein-PHB complexes are normally insoluble in chloroform, soluble in water and alkaline hypochlorite, and are converted to crotonic acid more slowly on heating in concentrated sulfuric acid. Our studies indicate that the majority of cellular PHB (over 80%) is located in cytoplasmic proteins, especially proteins of the ribosomal fraction. Western immunoblots, probed with polyclonal anti-PHB IgG, revealed a number of PHB-polypeptides having a wide range of molecular weights in all cell fractions. These results suggest that PHB is a fundamental constituent of cells that may have physiological functions in addition to facilitating ion transmembrane transport or serving as a carbon reserve. PMID- 8703034 TI - Identification and characterization of a novel human microsomal glutathione S transferase with leukotriene C4 synthase activity and significant sequence identity to 5-lipoxygenase-activating protein and leukotriene C4 synthase. AB - 5-Lipoxygenase-activating protein (FLAP) and leukotriene C4 (LTC4) synthase, two proteins involved in leukotriene biosynthesis, have been demonstrated to be 31% identical at the amino acid level. We have recently identified and characterized a novel member of the FLAP/LTC4 synthase gene family termed microsomal glutathione S-transferase II (microsomal GST-II). The open reading frame encodes a 16.6-kDa protein with a calculated pI of 10.4. Microsomal GST-II has 33% amino acid identity to FLAP, 44% amino acid identity to LTC4 synthase, and 11% amino acid identity to the previously characterized human microsomal GST (microsomal GST-I). Microsomal GST-II also has a similar hydrophobicity pattern to FLAP, LTC4 synthase, and microsomal GST-I. Fluorescent in situ hybridization mapped microsomal GST-II to chromosomal localization 4q28-31. Microsomal GST-II has a wide tissue distribution (at the mRNA level) and was specifically expressed in human liver, spleen, skeletal muscle, heart, adrenals, pancreas, prostate, testis, fetal liver, and fetal spleen. In contrast, microsomal GST-II mRNA expression was very low (when present) in lung, brain, placenta, and bone marrow. This differs from FLAP mRNA, which was detected in lung, various organs of the immune system, and peripheral blood leukocytes, and LTC4 synthase mRNA, which could not be detected in any tissues by Northern blot analysis. Microsomal GST-II and LTC4 synthase were expressed in a baculovirus insect cell system, and microsomes from Sf9 cells containing microsomal GST-II or LTC4 synthase were both found to catalyze the production of LTC4 from LTA4 and reduced glutathione. Microsomal GST-II also catalyzed the formation of another product, displaying a conjugated triene UV absorption spectra with a maximum at 283 nm, suggesting less catalytic stereospecificity compared with LTC4 synthase. Also, the apparent Km for LTA4 was higher for microsomal GST-II (41 microM) than LTC4 synthase (7 microM). In addition, unlike LTC4 synthase, microsomal GST-II was able to catalyze the conjugation of 1-chloro-2, 4-dinitrobenzene with reduced glutathione. Therefore, it is proposed that this novel membrane protein is a member of the microsomal glutathione S-transferase family, also including LTC4 synthase, with significant sequence identities to both LTC4 synthase and FLAP. PMID- 8703035 TI - Branching tubulogenesis but not scatter of madin-darby canine kidney cells requires a functional Grb2 binding site in the Met receptor tyrosine kinase. AB - Hepatocyte growth factor is a multifunctional cytokine that induces mitogenesis, motility, invasion, and branching tubulogenesis of several epithelial and endothelial cell lines in culture. The receptor for hepatocyte growth factor has been identified as the Met tyrosine kinase. To investigate the signaling pathways that are involved in these events, we have generated chimeric receptors containing the colony stimulating factor-1 receptor fused to the transmembrane and intracellular domains of the Met receptor. Madin-Darby canine kidney epithelial cells expressing the Met chimera dissociate scatter and form branching tubules in response to colony stimulating factor-1. From structure-function analyses, tyrosine residue 1356 within the carboxyl terminus of the Met receptor is critical for these events. The amino acid sequence downstream from tyrosine 1356 represents a consensus binding site for the Grb2 adaptor protein and forms a multisubstrate binding site for the p85 subunit of phosphatidylinositol 3-kinase, phospholipase Cgamma, and the Shc adaptor protein. To distinguish which of these signaling pathways are required, we generated a mutant receptor that selectively fails to associate with the Grb2 adaptor protein. Cells expressing this mutant receptor scattered but were unable to form branching tubules, indicating that a Grb2 binding site in the Met receptor is critical for morphogenic responses. PMID- 8703036 TI - Receptor-associated protein is a folding chaperone for low density lipoprotein receptor-related protein. AB - The 39-kDa receptor-associated protein (RAP) is a receptor antagonist that inhibits ligand interactions with the receptors that belong to the low density lipoprotein receptor gene family. Our previous studies have demonstrated that RAP interacts with the low density lipoprotein receptor-related protein (LRP) within the endoplasmic reticulum and prevents premature interaction of ligands with the receptor. To analyze whether RAP is also involved in the folding of LRP during receptor biosynthesis, we generated anchor-free, soluble minireceptors that represent each of the four putative ligand-binding domains of LRP (SLRP1, -2, -3, and -4, corresponding to the clusters with 2, 8, 10, and 11 cysteine-rich complement-type repeats, respectively). When these SLRPs were overexpressed by cell transfection, only SLRP1 was secreted. Little or no secretion was observed for SLRP2, -3, and -4. However, when RAP cDNA was cotransfected with SLRP2, -3, and -4 cDNAs, each of these SLRPs was secreted. The cellular retention of SLRPs in the absence of RAP coexpression appeared to be a result of the formation of SDS-resistant, oligomeric aggregates observed under nonreducing conditions. Such oligomers of the SLRPs likely resulted from formation of intermolecular disulfide bonds since they were reduced to monomers when analyzed under reducing conditions. The oligomers were formed not only among molecules of a given SLRP, but also between different SLRPs. The role of RAP in the process of LRP folding was shown by the reduction in aggregated SLRP oligomers upon RAP coexpression. A similar role of RAP in preventing the aggregation of newly synthesized receptor was also observed using membrane-containing minireceptor of LRP. Coimmunoprecipitation and ligand binding studies demonstrated that RAP binds avidly to SLRP2, -3, and -4, but not to SLRP1. These results suggest that these interactions may be important for proper folding of LRP by ensuring the formation of proper intradomain, but not intermolecular or interdomain, disulfide bonds. Thus, our results strongly suggest that, in addition to the prevention of premature binding of ligands to LRP, RAP also plays an important role in receptor folding. PMID- 8703037 TI - Differential regulation of activation-induced tyrosine phosphorylation and recruitment of SLP-76 to Vav by distinct isoforms of the CD45 protein-tyrosine phosphatase. AB - The CD45 family of transmembrane protein-tyrosine phosphatases plays a critical role in T cell activation signaling by regulating the tyrosine phosphorylation of protein-tyrosine kinases and their substrates. Multiple alternatively spliced CD45 isoforms, differing only in their extracellular domains, are differentially expressed by subsets of T cells with distinct functional repertoires. However, the physiological function of the various isoforms remains elusive. Using a novel panel of Jurkat T cell clones that uniquely express either the smallest (CD45(0)) or the largest (CD45(ABC)) isoform, we previously demonstrated CD45 isoform specific differences in interleukin-2 secretion and tyrosine phosphorylation of Vav. We now demonstrate differential activation-induced tyrosine phosphorylation of a 76-kDa Vav-associated protein (pp76) by cells expressing distinct CD45 isoforms. The tyrosine phosphorylation of Vav and associated pp76 follow parallel kinetics. pp76 interacts with the SH2 and SH3 domains of Vav. We have identified pp76 as SLP-76, a recently cloned Grb2-binding protein. After activation with anti-CD3, CD45(ABC) transfectants demonstrate increased tyrosine phosphorylation and physical association of SLP-76 with Vav compared to transfectants expressing CD45(0). These results establish a novel physical link between Vav and SLP-76 that is differentially regulated by CD45 isoform expression. PMID- 8703038 TI - The initiator element and proximal upstream sequences affect transcriptional activity and start site selection in the amyloid beta-protein precursor promoter. AB - The TATA-less human amyloid beta-protein precursor promoter contains an initiator element with the sequence CGTCA+1GTT. Primary transcriptional start sites were identified at positions +1 and -4. Deletion of the upstream activator elements APBbeta and APBalpha did not affect the selection of transcriptional start sites, although total transcriptional activity was reduced both in vitro and in vivo. Mutations within the initiator element shifted the transcriptional start sites and reduced transcriptional activity. Mutations between positions -6 and -35 changed the relative utilization of start sites +1 and -4 without affecting the total level of transcriptional activity. A 10-base pair deletion between position -40 and -31 increased in vitro transcriptional activity with a preeminent utilization of the start site at position -4. In contrast, a 20-base pair deletion between position -40 and -21 resulted in a reduction in transcriptional activity and in the primary utilization of the start site at position +1. Furthermore, transactivation by APBbeta and APBalpha was eliminated. DNase I footprinting provided evidence for the existence of two binding domains designated UE (position -12 to -30) and Inr (position +7 to -7). The positions of these binding domains are altered in mutations and deletions that affect transcriptional activity. PMID- 8703039 TI - Identification of key amino acids in a conserved cGMP-binding site of cGMP binding phosphodiesterases. A putative NKXnD motif for cGMP binding. AB - cGMP-binding phosphodiesterases contain two kinetically distinct cGMP-binding sites (a and b), and each site contains a conserved N(K/R)XnFX3DE sequence. N276A, K277A, K277R, D289A, and E290A mutants in the N276KX7FX3DE290 sequence of site a (higher affinity site) of bovine cGMP-binding, cGMP-specific phosphodiesterase (cGB-PDE or PDE5A) were expressed in High Five cells and purified. The cGMP-binding affinities of three mutants [K277A (Kd approximately 12 microM), D289A (Kd approximately 24 microM), and N276A (Kd approximately 60 microM)] were decreased in comparison with wild-type enzyme (Kd = 1.3 microM), which suggested an important role for Asn276, Lys277, and Asp289 in cGMP binding. These residues could be presented as a putative NKXnD motif, and their functions were predicted based on analogy with the canonical NKXD motif in GTP-binding proteins. No marked differences in catalytic functions such as specific activity, Km for cGMP, and IC50 for zaprinast or 3-isobutyl-1-methylxanthine were found among wild-type and mutant cGB-PDEs. This suggested that cGMP binding to site a does not influence the catalytic properties of cGB-PDE. PMID- 8703040 TI - Identification of a C-reactive protein binding site in two hepatic carboxylesterases capable of retaining C-reactive protein within the endoplasmic reticulum. AB - C-reactive protein (CRP) is normally synthesized by hepatocytes at relatively low rates and is retained within the endoplasmic reticulum (ER) via interaction with two carboxylesterases (termed gp60a and gp60b), which themselves are restricted to the ER by their COOH-terminal retention signals (HIEL and HTEL). During the acute phase response, an increase in CRP synthesis is accompanied by a decrease in its ER retention as a result of a decrease in the CRP binding affinity of gp60b. Our previous data indicated that the esterase active site, the CRP binding site, and the ER retention signal are functionally distinct. In the present studies, we have identified CRP-binding peptides produced by proteolytic cleavage of gp60a. The sequence shared by two CRP-binding peptides indicated that the CRP binding site of gp60a is contained within residues 477-499. These results were confirmed by expression of cDNAs coding for gp60a and b as bacterial fusion proteins. Fusion proteins containing the complete esterase COOH terminus bound CRP, whereas those truncated at residue 477 (or the homologous site in gp60b) did not. Based on the known crystal structures of three homologous hydrolases, the putative CRP-binding site of the gp60s is located on the surface and is physically distant from the esterase active site and the COOH-terminal ER retention signal. PMID- 8703041 TI - Activation of mitogen-activated protein (MAP) kinase pathway by pervanadate, a potent inhibitor of tyrosine phosphatases. AB - Rapid tyrosine phosphorylation of key cellular proteins is a crucial event in signal transduction. The regulatory role of protein-tyrosine phosphatases (PTPs) in this process was explored by studying the effects of a powerful PTP inhibitor, pervanadate, on the activation of the mitogen-activated protein (MAP) kinase cascade. Treatment of HeLa cells with pervanadate resulted in a marked inhibition of PTP activity, accompanied by a drastic increase in tyrosine phosphorylation of cellular proteins. The increased tyrosine phosphorylation coincided with the activation of the MAP kinase cascade as indicated by enzymatic activity assays of MEK (MAP kinase/ERK-kinase) and MAP kinase and gel mobility shift analyses of Raf 1 and MAP kinase. The activation was sustained but reversible. Upon removal of pervanadate, both tyrosine phosphorylation and MAP kinase activation declined to basal levels. Therefore, inhibition of PTP activity is sufficient per se to initiate a complete MAP kinase activation program. PMID- 8703042 TI - Chaperonin-promoted post-translational membrane insertion of a multispanning membrane protein lactose permease. AB - Using an in vitro membrane-free translation system from Escherichia coli, it is shown that chaperonin GroEL added cotranslationally interacts with newly synthesized lactose permease (LacY), a polytopic membrane protein, thereby preventing aggregation. Subsequently, when the isolated GroEL-LacY complex is incubated with inverted membrane vesicles, the permease is inserted into the membrane in a MgATP-dependent manner. Post-translational membrane insertion is also observed when aggregation of newly synthesized LacY is prevented by addition of the nonionic detergent n-dodecyl-beta,D-maltoside during translation in place of GroEL. No membrane integration occurs with right-side-out vesicles, indicating that LacY interacts specifically only with the cytosolic face of the membrane. Ligand thiodigalactoside protection against alkylation of the Cys-148 residue in the permease shows proper post-translational insertion. Moreover, limited proteolysis of soluble LacY either complexed with GroEL or in detergent indicates that the newly synthesized protein assumes a conformation that is comparable to that of native, membrane-embedded permease prior to insertion into the membrane. PMID- 8703043 TI - C/EBP-beta/LAP controls down-regulation of albumin gene transcription during liver regeneration. AB - Expression of the albumin gene in the liver is controlled by several liver enriched transcription factors. However, the mechanisms which contribute to its regulation during pathophysiological states, such as liver regeneration, are still little understood. In the present study we found that during liver regeneration down-regulation of albumin mRNA expression is transcriptionally controlled through a minimal element (nucleotide -170 to +22) of the albumin promoter and is observed mainly during the G1 phase of the cell cycle, while high levels of albumin expression are preserved at later time points. Decreased albumin mRNA levels correlate with a dramatic increase in nuclear expression of C/EBP-beta/LAP, a protein known to bind to the D site of the albumin promoter and also to be involved in cell cycle control. In contrast, nuclear expression of other factors such as HNF-1 or C/EBP-alpha, which also have been shown to transcriptionally control albumin expression, is either unchanged or slightly decreased. We show that pre- and post-translational mechanisms are involved in the higher nuclear expression of C/EBP-beta/LAP as early as 1 h after hepatectomy, which also leads to its increased binding toward the D site of the albumin promoter. Finally, in vitro transcription assays with liver nuclear extracts and recombinant C/EBP-beta/LAP demonstrate that C/EBP-beta/LAP can directly down-regulate transcription mediated by the minimal element of the albumin promoter. Additionally the inhibitory role of C/EBP-beta/LAP on the albumin minimal promoter could be confirmed by transfection experiments in hepatoma cells. These results indicate that C/EBP-beta/LAP, while enhancing transcription of cell cycle-related genes and controlling G1/S phase checkpoint, down-regulates a major liver function, i.e. albumin synthesis, to prepare the hepatocyte for entry into the cell cycle. PMID- 8703044 TI - The CCAAT-binding proteins CP1 and NF-I cooperate with ATF-2 in the transcription of the fibronectin gene. AB - We have previously proposed a molecular interaction between the liver factors that bind to the cyclic AMP response element (CRE) and CCAAT sites of the fibronectin (FN) gene based on the following evidence: (i) the close spacing of 20 base pairs between CRE and CCAAT elements is conserved in the FN genes from rats, mice, and humans; (ii) footprinting competitions showed that CRE oligonucleotides are able to detach both liver factors; (iii) CCAAT binding and transcriptional activity of liver extracts are reduced when the distance between the CRE and CCAAT elements is increased; and (iv) CCAAT-binding is stimulated by the addition of a liver extract fraction containing the CRE-binding factor ATF-2. This report provides binding and immunochemical evidence that nuclear factor I (CTF/NF-I) and CP1 (NF-Y or CBF) are the only liver factors that bind to the -150 CCAAT element of the FN gene, forming distinct complexes. We show that these factors bind less efficiently to the CCAAT site of a FN promoter in which the 170 CRE has been disrupted by site-directed mutagenesis and that each element contributes positively to the liver transcriptional activity assessed in vitro with a G-less cassette construct and in vivo by transfection of hepatoma cells with CAT constructs. Furthermore, using a method that combines UV cross-linking and immunoprecipitation, we show that antibodies specific to ATF-2 are able to specifically precipitate protein-protein-DNA complexes containing NF-I and CP1. This simple method preserves weak macromolecular interactions, avoiding the disruptive electrophoresis conditions of gel mobility shifts assays. PMID- 8703045 TI - RNA editing: a mechanism for gRNA-specified uridylate insertion into precursor mRNA. AB - In the mitochondria of trypanosomatid protozoa the precursors of messenger RNAs (pre-mRNAs) have their coding information remodeled by the site-specific insertion and deletion of uridylate (U) residues. Small trans-acting guide RNAs (gRNAs) supply the genetic information for this RNA editing. An in vitro system was developed to study the mechanism of U insertion into pre-mRNA. U-insertion editing occurs through a series of enzymatic steps that begin with gRNA-directed pre-mRNA cleavage. Inserted U's are derived from free uridine triphosphate and are added to the 3' terminus of a 5' pre-mRNA cleavage product. gRNA specifies edited RNA sequence at the subsequent ligation step by base pairing-mediated juxtaposition of the 3' cleavage product and the processed 5' cleavage product. gRNA/pre-mRNA chimeras, purported intermediates, seem to be abortive end products of the same reaction. PMID- 8703046 TI - Phase and morphology changes in lipid monolayers induced by SP-B protein and its amino-terminal peptide. AB - Both human lung surfactant protein, SP-B, and its amino-terminal peptide, SP-B1 25, inhibit the formation of condensed phases in monolayers of palmitic acid, resulting in a new fluid phase. This fluid phase forms a network, separating condensed-phase domains at coexistence. The network persists to high surface pressures, altering the nucleation, growth, and morphology of monolayer collapse structures, leading to lower surface tensions on compression and more reversible respreading on expansion. The network is stabilized by the low line tension between the fluid phase and the condensed phase as confirmed by the formation of "stripe" phases. PMID- 8703047 TI - Attomole protein characterization by capillary electrophoresis-mass spectrometry. AB - Electrospray ionization with an ultralow flow rate (30 kilodaltons) at a resolving power of approximately 60,000 for injections of 0.7 x 10(-18) to 3 x 10(-18) mole of 8- to 29-kilodalton proteins with errors of <1 dalton in molecular mass. Using a crude isolate from human blood, a value of 28,780.6 daltons (calculated, 28,780.4 daltons) was measured for carbonic anhydrase, representing 1 percent by weight of the protein in a single red blood cell. Dissociation of molecular ions from 9 x 10(-18) mole of carbonic anhydrase gave nine sequence-specific fragment ions, more data than required for unique retrieval of this enzyme from the protein database. PMID- 8703052 TI - Allelic diversity and gene genealogy at the self-incompatibility locus in the Solanaceae. AB - The self-incompatibility (S) locus of flowering plants offers an example of extreme polymorphism maintained by balancing selection. Estimates of recent and long-term effective population size (Ne) were determined for two solanaceous species by examination of S-allele diversity. Estimates of recent Ne in two solanaceous species differed by an order of magnitude, consistent with differences in the species' ecology. In one species, the evidence was consistent with historical population restriction despite a large recent Ne. In the other, no severe bottleneck was indicated over millions of years. Bottlenecks are integral to founder-event speciation, and loci that are subject to balancing selection can be used to evaluate the frequency of this mode of speciation. PMID- 8703053 TI - Forskolin stimulation of water and cation permeability in aquaporin 1 water channels. AB - Aquaporin 1, a six-transmembrane domain protein, is a water channel present in many fluid-secreting and -absorbing cells. In Xenopus oocytes injected with aquaporin 1 complementary RNA, the application of forskolin or cyclic 8-bromo- adenosine 3',5'-monophosphate increased membrane permeability to water and triggered a cationic conductance. The cationic conductance was also induced by direct injection of protein kinase A (PKA) catalytic subunit, reduced by the kinase inhibitor H7, and blocked by HgCl2, an inhibitor of aquaporin 1. The cationic permeability of the aquaporin 1 channel is activated by a cyclic adenosine monophosphate-dependent mechanism that may involve direct or indirect phosphorylation by PKA. PMID- 8703055 TI - Identification of an estrogen response element activated by metabolites of 17beta estradiol and raloxifene. AB - 17beta-Estradiol modulates gene transcription through the estrogen receptor and the estrogen response element in DNA. The human transforming growth factor-beta3 gene was shown to be activated by the estrogen receptor in the presence of estrogen metabolites or estrogen antagonists. Activation was mediated by a polypurine sequence, termed the raloxifene response element, and did not require the DNA binding domain of the estrogen receptor. Interaction of the estrogen receptor with the raloxifene response element appears to require a cellular adapter protein. The observation that individual estrogens modulate multiple DNA response elements may explain the tissue-selective estrogen agonist or antagonist activity of compounds such as raloxifene. PMID- 8703054 TI - Luteinizing hormone deficiency and female infertility in mice lacking the transcription factor NGFI-A (Egr-1). AB - The immediate-early transcription factor NGFI-A (also called Egr-1, zif/268, or Krox-24) is thought to couple extracellular signals to changes in gene expression. Although activins and inhibins regulate follicle-stimulating hormone (FSH) synthesis, no factor has been identified that exclusively regulates luteinizing hormone (LH) synthesis. An analysis of NGFI-A-deficient mice derived from embryonic stem cells demonstrated female infertility that was secondary to LH-beta deficiency. Ovariectomy led to increased amounts of FSH-beta but not LH beta messenger RNA, which suggested a pituitary defect. A conserved, canonical NGFI-A site in the LH-beta promoter was required for synergistic activation by NGFI-A and steroidogenic factor-1 (SF-1). NGFI-A apparently influences female reproductive capacity through its regulation of LH-beta transcription. PMID- 8703056 TI - Tourette syndrome: prediction of phenotypic variation in monozygotic twins by caudate nucleus D2 receptor binding. AB - Tourette syndrome, a chronic tic disorder with autosomal dominant inheritance, exhibits considerable phenotypic variability even within monozygotic twin pairs. The origins of this variability remain unclear. Recent findings have implicated the caudate nucleus as a locus of pathology, and pharmacological evidence supports dopaminergic involvement. Within monozygotic twins discordant for Tourette syndrome severity, differences in D2 dopamine receptor binding in the head of the caudate nucleus predicted differences in phenotypic severity (r = 0.99); this relation was not observed in putamen. These data may link Tourette syndrome with a spectrum of neuropsychiatric disorders that involve associative striatal circuitry. PMID- 8703057 TI - KUZ, a conserved metalloprotease-disintegrin protein with two roles in Drosophila neurogenesis. AB - During neurogenesis in Drosophila both neurons and nonneuronal cells are produced from a population of initially equivalent cells. The kuzbanian (kuz) gene described here is essential for the partitioning of neural and nonneuronal cells during development of both the central and peripheral nervous systems in Drosophila. Mosaic analyses indicated that kuz is required for cells to receive signals inhibiting the neural fate. These analyses further revealed that the development of a neuron requires a kuz-mediated positive signal from neighboring cells. The kuz gene encodes a metalloprotease-disintegrin protein with a highly conserved bovine homolog, raising the possibility that kuz homologs may act in similar processes during mammalian neurogenesis. PMID- 8703058 TI - Modulation of virulence factor expression by pathogen target cell contact. AB - Upon contact with the eukaryotic cell, Yersinia pseudotuberculosis increased the rate of transcription of virulence genes (yop), as determined by in situ monitoring of light emission from individual bacteria expressing luciferase under the control of the yopE promoter. The microbe-host interaction triggered export of LcrQ, a negative regulator of Yop expression, via the Yop-type III secretion system. The intracellular concentration of LcrQ was thereby lowered, resulting in increased expression of Yops. These results suggest a key role for the type III secretion system of pathogenic bacteria to coordinate secretion with expression of virulence factors after physical contact with the target cell. PMID- 8703059 TI - Induction of gene expression in Escherichia coli after pilus-mediated adherence. AB - The induction of cascades of virulence factors after contact between bacteria and host cells was investigated. P-pili mediate the binding of uropathogenic Escherichia coli to its host cell receptor. After P-pili binding there was transcriptional activation of a sensor-regulator protein that is essential for the bacterial iron-starvation response. An insertion mutation of the sensor regulator gene eliminated the ability of uropathogenic E. coli to produce siderophores and their iron-regulated membrane receptors, thereby abolishing their ability to grow in urine. These results suggest that P-pilus-mediated attachment may be an important part of the sensor-regulatory process involved in uropathogenic E. coli urinary tract infection. PMID- 8703060 TI - Pycnodysostosis, a lysosomal disease caused by cathepsin K deficiency. AB - Pycnodysostosis, an autosomal recessive osteochondrodysplasia characterized by osteosclerosis and short stature, maps to chromosome 1q21. Cathepsin K, a cysteine protease gene that is highly expressed in osteoclasts, localized to the pycnodysostosis region. Nonsense, missense, and stop codon mutations in the gene encoding cathepsin K were identified in patients. Transient expression of complementary DNA containing the stop codon mutation resulted in messenger RNA but no immunologically detectable protein. Thus, pycnodysostosis results from gene defects in a lysosomal protease with highest expression in osteoclasts. These findings suggest that cathepsin K is a major protease in bone resorption, providing a possible rationale for the treatment of disorders such as osteoporosis and certain forms of arthritis. PMID- 8703061 TI - A protein farnesyl transferase involved in abscisic acid signal transduction in Arabidopsis. AB - The hormone abscisic acid (ABA) modulates a variety of developmental processes and responses to environmental stress in higher plants. A collection of mutations, designated era, in Arabidopsis thaliana that confer an enhanced response to exogenous ABA includes mutations in the Era1 gene, which encodes the beta subunit of a protein farnesyl transferase. In yeast and mammalian systems, farnesyl transferases modify several signal transduction proteins for membrane localization. The era1 mutants suggest that a negative regulator of ABA sensitivity must be acted on by a farnesyl transferase to function. PMID- 8703062 TI - CD2: an exception to the immunoglobulin superfamily concept? PMID- 8703065 TI - A silicate inclusion in Puente del Zacate, a IIIA iron meteorite. AB - The IIIA and IIIB iron meteorites are considered to have formed in the cores of asteroids. A silicate inclusion within the IIIA meteorite Puente del Zacate consisting of olivine (Fa4), low-calcium pyroxene (Fs6Wo1), chromium diopside (Fs3Wo47), plagioclase (An14Or4), graphite, troilite, chromite, daubreelite, and iron metal resembles inclusions in IAB iron meteorites. The oxygen isotopic composition of the Puente del Zacate inclusion is like chromite and phosphate inclusions in other IIIA and IIIB irons. The Puente del Zacate inclusion may have been derived from the lower mantle of the IIIAB parent asteroid. PMID- 8703069 TI - Antagonistic interactions between wingless and decapentaplegic responsible for dorsal-ventral pattern in the Drosophila Leg. AB - Subdivision of the limb primordia of Drosophila into anterior and posterior compartments triggers cell interactions that pattern the legs and wings. A comparable compartment-based mechanism is used to pattern the dorsal-ventral axis of the wing. Evidence is presented here for a mechanism based on cell interaction, rather than on compartment formation, that distinguishes dorsal from ventral in the leg. Mutual repression by Wingless and Decapentaplegic signaling systems generates a stable regulatory circuit by which each gene maintains its own expression in a spatially restricted domain. Compartment-independent patterning mechanisms may be used by other organisms during development. PMID- 8703070 TI - Purification and molecular cloning of Plx1, a Cdc25-regulatory kinase from Xenopus egg extracts. AB - Cdc2, the cyclin-dependent kinase that controls mitosis, is negatively regulated by phosphorylation on its threonine-14 and tyrosine-15 residues. Cdc25, the phosphatase that dephosphorylates both of these residues, undergoes activation and phosphorylation by multiple kinases at mitosis. Plx1, a kinase that associates with and phosphorylates the amino-terminal domain of Cdc25, was purified extensively from Xenopus egg extracts. Cloning of its complementary DNA revealed that Plx1 is related to the Polo family of protein kinases. Recombinant Plx1 phosphorylated Cdc25 and stimulated its activity in a purified system. Cdc25 phosphorylated by Plx1 reacted strongly with MPM-2, a monoclonal antibody to mitotic phosphoproteins. These studies indicate that Plx1 may participate in control of mitotic progression. PMID- 8703071 TI - A model of host-microbial interactions in an open mammalian ecosystem. AB - The maintenance and significance of the complex populations of microbes present in the mammalian intestine are poorly understood. Comparison of conventionally housed and germ-free NMRI mice revealed that production of fucosylated glycoconjugates and an alpha1, 2-fucosyltransferase messenger RNA in the small intestinal epithelium requires the normal microflora. Colonization of germ-free mice with Bacteroides thetaiotaomicron, a component of this flora, restored the fucosylation program, whereas an isogenic strain carrying a transposon insertion that disrupts its ability to use L-fucose as a carbon source did not. Simplified models such as this should aid the study of open microbial ecosystems. PMID- 8703072 TI - Time-sensitive reversal of hyperplasia in transgenic mice expressing SV40 T antigen. AB - The role of viral oncoprotein expression in the maintenance of cellular transformation was examined as a function of time through controlled expression of simian virus 40 T antigen (TAg). Expression of TAg in the submandibular gland of transgenic mice from the time of birth induced cellular transformation and extensive ductal hyperplasia by 4 months of age. The hyperplasia was reversed when TAg expression was silenced for 3 weeks. When TAg expression was silenced after 7 months, however, the hyperplasia persisted even though TAg was absent. Although the polyploidy of ductal cells could be reversed at 4 months of age, cells at 7 months of age remained polyploid even in the absence of TAg. These results support a model of time-dependent multistep tumorigenesis, in which virally transformed cells eventually lose their dependence on the viral oncoprotein for maintenance of the transformed state. PMID- 8703073 TI - Correction of the mutation responsible for sickle cell anemia by an RNA-DNA oligonucleotide. AB - A chimeric oligonucleotide composed of DNA and modified RNA residues was used to direct correction of the mutation in the hemoglobin betaS allele. After introduction of the chimeric molecule into lymphoblastoid cells homozygous for the betaS mutation, there was a detectable level of gene conversion of the mutant allele to the normal sequence. The efficient and specific conversion directed by chimeric molecules may hold promise as a therapeutic method for the treatment of genetic diseases. PMID- 8703074 TI - Enhanced protein C activation and inhibition of fibrinogen cleavage by a thrombin modulator. AB - A modulator of the enzymatic activity of human thrombin, designated LY254603, was identified that enhances the thrombin-catalyzed generation of the anticoagulant factor activated protein C, yet inhibits thrombin-dependent fibrinogen clotting. By means of mutant substrates, it was shown that LY254603 mediates the change in enzymatic substrate specificity through an alteration in thrombin's S3 substrate recognition site, a mechanism that appeared to be independent of allosteric changes induced by either sodium ions or by thrombomodulin. This compound may represent the prototype of a class of agents that specifically modulates the balance between thrombin's procoagulant and anticoagulant functions. PMID- 8703075 TI - Crystal structure of the Aequorea victoria green fluorescent protein. AB - The green fluorescent protein (GFP) from the Pacific Northwest jellyfish Aequorea victoria has generated intense interest as a marker for gene expression and localization of gene products. The chromophore, resulting from the spontaneous cyclization and oxidation of the sequence -Ser65 (or Thr65)-Tyr66-Gly67-, requires the native protein fold for both formation and fluorescence emission. The structure of Thr65 GFP has been determined at 1.9 angstrom resolution. The protein fold consists of an 11-stranded beta barrel with a coaxial helix, with the chromophore forming from the central helix. Directed mutagenesis of one residue adjacent to the chromophore, Thr203, to Tyr or His results in significantly red-shifted excitation and emission maxima. PMID- 8703076 TI - Bimodal interaction of coatomer with the p24 family of putative cargo receptors. AB - Cytoplasmic domains of members of the p24 family of putative cargo receptors were shown to bind to coatomer, the coat protein of COPI-coated transport vesicles. Domains that contained dilysine endoplasmic reticulum retrieval signals bound the alpha-, beta'-, and epsilon-COP subunits of coatomer, whereas other p24 domains bound the beta-, gamma-, and zeta-COP subunits and required a phenylalanine containing motif. Transit of a CD8-p24 chimera from the endoplasmic reticulum through the Golgi complex was slowed when the phenylalanine motif was mutated, suggesting that this motif may function as an anterograde transport signal. The either-or bimodal binding of coatomer to p24 tails suggests models for how coatomer can potentially package retrograde-directed and anterograde-directed cargo into distinct COPI-coated vesicles. PMID- 8703077 TI - A neostriatal habit learning system in humans. AB - Amnesic patients and nondemented patients with Parkinson's disease were given a probabilistic classification task in which they learned which of two outcomes would occur on each trial, given the particular combination of cues that appeared. Amnesic patients exhibited normal learning of the task but had severely impaired declarative memory for the training episode. In contrast, patients with Parkinson's disease failed to learn the probabilistic classification task, despite having intact memory for the training episode. This double dissociation shows that the limbic-diencephalic regions damaged in amnesia and the neostriatum damaged in Parkinson's disease support separate and parallel learning systems. In humans, the neostriatum (caudate nucleus and putamen) is essential for the gradual, incremental learning of associations that is characteristic of habit learning. The neostriatum is important not just for motor behavior and motor learning but also for acquiring nonmotor dispositions and tendencies that depend on new associations. PMID- 8703078 TI - A requirement for local protein synthesis in neurotrophin-induced hippocampal synaptic plasticity. AB - Two neurotrophic factors, brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NT-3), are able to produce a long-lasting enhancement of synaptic transmission in the hippocampus. Unlike other forms of plasticity, neurotrophin induced plasticity exhibited an immediate requirement for protein synthesis. Plasticity in rat hippocampal slices in which the synaptic neuropil was isolated from the principal cell bodies also required early protein synthesis. Thus, the neurotrophins may stimulate the synthesis of proteins in either axonal or dendritic compartments, allowing synapses to exert local control over the complement of proteins expressed at individual synaptic sites. PMID- 8703079 TI - CRINKLY4: A TNFR-like receptor kinase involved in maize epidermal differentiation. AB - The maize crinkly4 (cr4) mutation affects leaf epidermis differentiation such that cell size and morphology are altered, and surface functions are compromised, allowing graft-like fusions between organs. In the seed, loss of cr4 inhibits aleurone formation in a pattern that reflects the normal progression of differentiation over the developing endosperm surface. The cr4 gene was isolated by transposon tagging and found to encode a putative receptor kinase. The extracellular domain contains a cysteine-rich region similar to the ligand binding domain in mammalian tumor necrosis factor receptors (TNFRs) and seven copies of a previously unknown 39-amino acid repeat. The results suggest a role for cr4 in a differentiation signal. PMID- 8703080 TI - Similarity of a chromatic adaptation sensor to phytochrome and ethylene receptors. AB - Complementary chromatic adaptation in cyanobacteria acts through photoreceptors to control the biosynthesis of light-harvesting complexes. The mutant FdBk, which appears black, cannot chromatically adapt and may contain a lesion in the apparatus that senses light quality. The complementing gene identified here, rcaE, encodes a deduced protein in which the amino-terminal region resembles the chromophore attachment domain of phytochrome photoreceptors and regions of plant ethylene receptors; the carboxyl- terminal half is similar to the histidine kinase domain of two-component sensor kinases. PMID- 8703081 TI - Exploring phenotype space through neutral evolution. AB - RNA secondary-structure folding algorithms predict the existence of connected networks of RNA sequences with identical secondary structures. Fitness landscapes that are based on the mapping between RNA sequence and RNA secondary structure hence have many neutral paths. A neutral walk on these fitness landscapes gives access to a virtually unlimited number of secondary structures that are a single point mutation from the neutral path. This shows that neutral evolution explores phenotype space and can play a role in adaptation. PMID- 8703082 TI - Class III POU genes: generation of homopolymeric amino acid repeats under GC pressure in mammals. AB - The class III POU transcription factor genes play an important role in the nervous system. Comparison of their entire amino acid sequences disclosed a remarkable feature of particular mammalian class III POU genes. Alanine, glycine, and proline repeats were present in the mammalian Brain-1 gene, whereas most of these repeats were absent in the nonmammalian homologue. The mammalian Brain-2 gene had alanine, glycine, proline, and glutamine repeats, which were missing in the nonmammalian homologue. The mammalian Scip gene had alanine, glycine, proline, and histidine repeats, but the nonmammalian homologue completely lacked these repeats. In contrast, the mammalian Brain-4 gene had no amino acid repeats like its nonmammalian homologue. The mammalian genes containing the characteristic amino acid repeats had another feature, higher GC content. We found a positive correlation between the GC content and the amino acid repeat ratio. Those amino acids were encoded by triplet codons with relatively high GC content. These results suggest that the GC pressure has facilitated generation of the homopolymeric amino acid repeats. PMID- 8703083 TI - Mu opioid receptor-like sequences are present throughout vertebrate evolution. AB - The sequence of the mu opioid receptor is highly conserved among human, rat, and mouse. In order to gain insights into the evolution of the mu opioid receptor, polymerase chain reaction (PCR) was used to screen genomic DNA from a number of different species using degenerate oligonucleotides which recognize a highly conserved region. DNA was assayed from representative species of both the protostome and deuterostome branches of the metazoan phylogenetic tree. Mu opioid receptor-like sequences were found in all vertebrate species that were analyzed. These species included bovine, chicken, bullfrog, striped bass, thresher shark, and Pacific hagfish. However, no mu opioid receptor-like sequences were detected from protostomes or from any invertebrates. The PCR results demonstrate that the region of the mu opioid receptor gene between the first intracellular loop and the third transmembrane domain (TM3) has been highly conserved during evolution and that mu opioid receptor-like sequences are present in the earliest stages of vertebrate evolution. Additional opioid receptor-like sequence was obtained from mRNA isolated from Pacific hagfish brain using rapid amplification of cDNA ends (RACE). The sequence of the Pacific hagfish was most homologous with the human mu opioid receptor (72% at the amino acid level between intracellular loop 1 and transmembrane domain 6) although over the same region high homology was also observed with the delta opioid receptor (69%), the kappa receptor (63%), and opioid receptor-like (ORL1) (59%). The hagfish sequence showed low conservation with the mammalian opioid receptors in the first and second extracellular loops but high conservation in the transmembrane and intracellular domains. PMID- 8703084 TI - Evolution of promoter sequences: elements of a canonical promoter for prespore genes of Dictyostelium. AB - An attempt is made to define a minimal prespore promoter which contains all elements essential for correct regulation of expression of a prespore gene. The prespore genes of Dictyostelium are coregulated during development. Most begin transcription at the same early stage, and activity of all is restricted to prespore tissue during the later slug stage. Sequences 5' to the coding sequences of eight prespore genes were searched for all elements proposed to control transcription and for new elements. The meaningfulness of occurrences of elements and pairs of elements in prespore promoters was evaluated by comparison with frequencies of occurrences in promoters of other, nonprespore genes. These comparisons resulted in definition of a canonical prespore promoter, a stretch of about 200 nucleotides containing at least one of each of three elements. Certain limitations were found on the spacing of elements. Orientation of elements with respect to each other appeared unrestricted. All elements often occurred in multiple copies. This structure suggests that individual copies of each element are not conserved during evolution, but instead continually appear and disappear. PMID- 8703085 TI - Common features of analogous replacement histone H3 genes in animals and plants. AB - Phylogenetic analysis of histone H3 protein sequences demonstrates the independent origin of the replacement histone H3 genes in animals and in plants. Multiple introns in the replacement histone H3 genes of animals in a pattern distinct from that in plant replacement H3 genes supports this conclusion. It is suggested that replacement H3 genes arose at the same time that, independently, multicellular forms of animals and of plants evolved. Judged by the degree of invariant and functionally constrained amino acid positions, histones H3 and H4, which form together the tetramer kernel of the nucleosome, have co-evolved with equal rates of sequence divergence. Residues 31 and 87 in histone H3 are the only residues that consistently changed across each gene duplication event that created functional replacement histone H3 variant forms. Once changed, these residues have remained invariant across divergent speciation. This suggests that they are required to allow replacement histone H3 to participate in the assembly of nucleosomes in non-S-phase cells. The abundant occurrence of polypyrimidine sequences in the introns of all replacement H3 genes, and the replacement of an intron by a polypyrimidine motif upstream of the alfalfa replacement H3 gene, suggests a function. It is speculated that they may contribute to the characteristic cell-cycle-independent pattern of replacement histone H3 genes by binding nucleosome-excluding proteins. PMID- 8703087 TI - A relationship between GC content and coding-sequence length. AB - Since base composition of translational stop codons (TAG, TAA, and TGA) is biased toward a low G+C content, a differential density for these termination signals is expected in random DNA sequences of different base compositions. The expected length of reading frames (DNA segments of sense codons flanked by in-phase stop codons) in random sequences is thus a function of GC content. The analysis of DNA sequences from several genome databases stratified according to GC content reveals that the longest coding sequences-exons in vertebrates and genes in prokaryotes-are GC-rich, while the shortest ones are GC-poor. Exon lengthening in GC-rich vertebrate regions does not result, however, in longer vertebrate proteins, perhaps because of the lower number of exons in the genes located in these regions. The effects on coding-sequence lengths constitute a new evolutionary meaning for compositional variations in DNA GC content. PMID- 8703086 TI - Phylogenetic relationships of annelids, molluscs, and arthropods evidenced from molecules and morphology. AB - Annelids and arthropods have long been considered each other's closest relatives, as evidenced by similarities in their segmented body plans. An alternative view, more recently advocated by investigators who have examined partial 18S ribosomal RNA data, proposes that annelids, molluscs, and certain other minor phyla with trochophore larva stages share a more recent common ancestor with one another than any do with arthropods. The two hypotheses are mutually exclusive in explaining spiralian relationships. Cladistic analysis of morphological data does not reveal phylogentic relationships among major spiralian taxa but does suggest monophyly for both the annelids and molluscs. Distance and maximum-likelihood analyses of 18S rRNA gene sequences from major spiralian taxa suggest a sister relationship between annelids and molluscs and provide a clear resolution within the major groups of the spiralians. The parsimonious tree based on molecular data, however, indicates a sister relationship of the Annelida and Bivalvia, and an earlier divergence of the Gastropoda than the Annelida-Bivalvia clade. To test further hypotheses on the phylogenetic relationships among annelids, molluscs, and arthropods, and the ingroup relationships within the major spiralian taxa, we combine the molecular and morphological data sets and subject the combined data matrix to parsimony analysis. The resulting tree suggests that the molluscs and annelids form a monophyletic lineage and unites the molluscan taxa to a monophyletic group. Therefore, the result supports the Eutrochozoa hypothesis and the monophyly of molluscs, and indicates early acquisition of segmented body plans in arthropods. PMID- 8703089 TI - Heterogeneity of the glutathione transferase genes encoding enzymes responsible for insecticide degradation in the housefly. AB - One of the four glutathione-S-transferases (GST) that is overproduced in the insecticide-resistant Cornell-R strain of the housefly (Musca domestica) produces an activity that degrades the insecticide dimethyl parathion and conjugates glutathione to lindane. In earlier work, it was shown that the resistant Cornell R carries an amplification, probably a duplication, of one or more of its GST loci and that this amplification is directly related to resistance. Using polymerase chain reaction (PCR) amplification with genomic DNA, multiple copies of the gene encoding the parathion-degrading activity (called MdGst-3) were subcloned from both the ancestral, insecticide-susceptible strain BPM and from the insecticide-resistant Cornell-R. In BPM, three different MdGst-3 genes were identified while in Cornell-R, 12 different MdGst-3 sequences were found that, though closely related to ancestral genes, had diverged by a few nucleotides. This diversity in MdGst-3 genomic sequences in Cornell-R is reflected in the expressed sequences, as sampled through a cDNA bank. Population heterozygosity cannot account for these multiple GST genes. We suggest that selection for resistance to insecticides has resulted in not only amplification of the MdGst-3 genes but also in the divergence of sequence between the amplified copies. PMID- 8703088 TI - Actin gene structure in two Artemia species, A. franciscana and A. parthenogenetica. AB - Genomic clones coding for actin have been isolated from two species of the crustacean Artemia, A. parthenogenetica and A. franciscana. The Act211 isoform gene was isolated from A. parthenogenetica, and the two other isoform genes, Act302 and Act403, were isolated from A. franciscana. The comparison of the nucleotide sequence of genomic and cDNA clones showed an interspecific divergence of 4% in translated and 6.1% in untranslated regions. However, the establishment of the partial structure of the Act211 gene in A. franciscana and of the Act302 gene in A. parthenogenetica suggests their similarity in the two species. The Act211 gene is divided into four exons, the Act302 gene into six exons, and the Act403 gene into seven exons. The three genes have introns in the 5' untranslated region and between codons 41 and 42. The Act211 and 403 genes have one common intron in codon 168. The Act302 and 403 genes have common introns between codons 121-122, 246-247, and within codon 301. While introns in the 5' untranslated region and between codons 41-42 and 121-122 are present in many organisms, the introns in positions 168 and 246-247 had only been found previously in actin genes from the nematode Onchocerca volvulus and the green alga Volvox carterii, respectively. The intron in position 301 had not been reported before. The transcription initiation sites of these three genes as well as the nucleotide sequences of the promoter regions have been also determined. PMID- 8703090 TI - Duplication and divergence of the genes of the alpha-esterase cluster of Drosophila melanogaster. AB - The alpha-esterase cluster of D. melanogaster contains 11 esterase genes dispersed over 60 kb. Embedded in the cluster are two unrelated open reading frames that have sequence similarity with genes encoding ubiquitin-conjugating enzyme and tropomyosin. The esterase amino acid sequences show 37-66% identity with one another and all but one have all the motifs characteristic of functional members of the carboxyl/cholinesterase multigene family. The exception has several frameshift mutations and appears to be a pseudogene. Patterns of amino acid differences among cluster members in relation to generic models of carboxyl/cholinesterase protein structure are broadly similar to those among other carboxyl/cholinesterases sequenced to date. However the alpha-esterases differ from most other members of the family in: their lack of a signal peptide; the lack of conservation in cysteines involved in disulfide bridges; and in four indels, two of which occur in or adjacent to regions that align with proposed substrate-binding sites of other carboxyl/cholinesterases. Phylogenetic analyses clearly identify three simple gene duplication events within the cluster. The most recent event involved the pseudogene which is located in an intron of another esterase gene. However, relative rate tests suggest that the pseudogene remained functional after the duplication event and has become inactive relatively recently. The distribution of indels also suggests a deeper node in the gene phylogeny that separates six genes at the two ends of the cluster from a block of five in the middle. PMID- 8703092 TI - Evolution of disintegrin cysteine-rich and mammalian matrix-degrading metalloproteinases: gene duplication and divergence of a common ancestor rather than convergent evolution. AB - The evolution of the Metalloproteinase Disintegrin Cysteine-rich (MDC) gene family and that of the mammalian Matrix-degrading Metalloproteinases (MMPs) are compared. The alignment of snake venom and mammalian MDC and MMP precursor sequences generated a phylogenetic tree that grouped these proteins mainly according to their function. Based on this observation, a common ancestry is suggested for mammalian and snake venom MDCs; it is also possible that gene duplication of the already-assembled domain structure, followed by divergence of the copies, may have significantly contributed to the evolution of the functionally diverse MDC proteins. The data also suggest that the structural resemblance of the zinc-binding motif of venom MDCs and MMPs may best be explained by common ancestry and conservation of the proteolytic motifs during the divergence of the proteins rather than through convergent evolution. PMID- 8703091 TI - Transcription in the early diverging eukaryote Trichomonas vaginalis: an unusual RNA polymerase II and alpha-amanitin-resistant transcription of protein-coding genes. AB - We have examined transcription in an early diverging eukaryote by analyzing the effect of the fungus-derived toxin alpha-amanitin on the transcription of protein coding genes of the protist Trichomonas vaginalis. In contrast to that typical in eukaryotes, the RNA polymerase that transcribes T. vaginalis protein-coding genes is relatively resistant to alpha-amanitin (50% inhibition = 250 microg alpha amanitin/ml). We have also characterized the gene encoding the largest subunit of RNA polymerase II, the subunit that binds alpha-amanitin. This protein is 41% identical to the mouse RNA polymerase II. Sequence analysis of the 50-amino-acid region thought to bind alpha-amanitin shows that this region of the trichomonad RNA polymerase II lacks many of the conserved amino acids present in the putative binding site, in agreement with the observed insensitivity to this inhibitor. Similar to other RNA polymerase IIs analyzed from ancient eukaryotes, the T. vaginalis RNA polymerase II lacks the typical heptapeptide (Tyr-Ser-Pro-Thr-Ser Pro-Ser) repeat carboxyl-terminal domain (CTD) that is a hallmark of higher eukaryotic RNA polymerase IIs. The trichomonad enzyme, however, does contain a short modified CTD that is rich in the amino acid residues that compose the repeat. These data suggest that T. vaginalis protein-coding genes are transcribed by a RNA polymerase II that is relatively insensitive to alpha-amanitin and that differs from typical eukaryotic RNA polymerase IIs as it lacks a heptapeptide repeated CTD. PMID- 8703093 TI - Gene structure of a chlorophyll a/c-binding protein from a brown alga: presence of an intron and phylogenetic implications. AB - A Laminaria saccharina genomic library in the phage EMBL 4 was used to isolate and sequence a full-length gene encoding a fucoxanthin-chlorophyll a/c-binding protein. Contrary to diatom homologues, the coding sequence is interrupted by an intron of about 900 bp which is located in the middle of the transit peptide. The deduced amino acid sequence of the mature protein is very similar to those of related proteins from Macrocystis pyrifera (Laminariales) and, to a lesser extent, to those from diatoms and Chrysophyceae. Seven of the eight putative chlorophyll-binding amino acids determined in green plants are also present. Alignments of different sequences related to the light-harvesting proteins (LHC) demonstrate a structural similarity among the three transmembrane helices and suggest a unique ancestral helix preceded by two beta-turns. The beta-turns are conserved in front of the second helices of the chlorophyll a/c proteins more so than in chlorophyll a/b proteins. Phylogenetic trees generated from sequence data indicate that fucoxanthin-chlorophyll-binding proteins diverged prior to the separation of photosystem I and photosystem II LHC genes of green plants. Among the fucoxanthin-containing algae, LHC I or II families could not be distinguished at this time. PMID- 8703094 TI - Molecular systematics of the Drosophila hydei subgroup as inferred from mitochondrial DNA sequences. AB - The phylogeny of the Drosophila hydei subgroup, which is a member of the D. repleta species group, was inferred from 1,515 base pairs of mitochondrial DNA sequence of the cytochrome oxidase subunits I, II, and III. Four of the seven species in the subgroup were examined, which are placed into two taxonomic complexes: the D. bifurca complex (D. bifurca and D. nigrohydei) and the D. hydei complex (D. hydei and D. eohydei). Both complexes appear to be monophyletic, although the D. bifurca complex is only weakly supported. The evolution of chromosomal change, interspecific crossability, sperm gigantism, and divergence times of the subgroup is discussed in a phylogenetic context. PMID- 8703095 TI - Molecular evidence for Acanthocephala as a subtaxon of Rotifera. AB - Rotifers are free-living animals usually smaller than 1 mm that possess a characteristic wheel organ. Acanthocephalans (thorny-headed worms) are larger endoparasitic animals that use vertebrates and arthropods to complete their life cycle. The taxa Acanthocephala and Rotifera are considered separate phyla, often within the taxon Aschelminthes. We have reexamined the relationship between Rotifera and Acanthocephala using 18S rRNA gene sequences. Our results conclusively show that Acanthocephala is the sister group of the rotifer class Bdelloidea. Rotifera was nonmonophyletic in all molecular analyses, which supports the hypothesis that the Acanthocephala represent a taxon within the phylum Rotifera and not a separate phylum. These results agree with a previous cladistic study of morphological characters. PMID- 8703096 TI - The puzzle of the Krebs citric acid cycle: assembling the pieces of chemically feasible reactions, and opportunism in the design of metabolic pathways during evolution. AB - The evolutionary origin of the Krebs citric acid cycle has been for a long time a model case in the understanding of the origin and evolution of metabolic pathways: How can the emergence of such a complex pathway be explained? A number of speculative studies have been carried out that have reached the conclusion that the Krebs cycle evolved from pathways for amino acid biosynthesis, but many important questions remain open: Why and how did the full pathway emerge from there? Are other alternative routes for the same purpose possible? Are they better or worse? Have they had any opportunity to be developed in cellular metabolism evolution? We have analyzed the Krebs cycle as a problem of chemical design to oxidize acetate yielding reduction equivalents to the respiratory chain to make ATP. Our analysis demonstrates that although there are several different chemical solutions to this problem, the design of this metabolic pathway as it occurs in living cells is the best chemical solution: It has the least possible number of steps and it also has the greatest ATP yielding. Study of the evolutionary possibilities of each one-taking the available material to build new pathways-demonstrates that the emergence of the Krebs cycle has been a typical case of opportunism in molecular evolution. Our analysis proves, therefore, that the role of opportunism in evolution has converted a problem of several possible chemical solutions into a single-solution problem, with the actual Krebs cycle demonstrated to be the best possible chemical design. Our results also allow us to derive the rules under which metabolic pathways emerged during the origin of life. PMID- 8703097 TI - Probability distribution of molecular evolutionary trees: a new method of phylogenetic inference. AB - A new method is presented for inferring evolutionary trees using nucleotide sequence data. The birth-death process is used as a model of speciation and extinction to specify the prior distribution of phylogenies and branching times. Nucleotide substitution is modeled by a continuous-time Markov process. Parameters of the branching model and the substitution model are estimated by maximum likelihood. The posterior probabilities of different phylogenies are calculated and the phylogeny with the highest posterior probability is chosen as the best estimate of the evolutionary relationship among species. We refer to this as the maximum posterior probability (MAP) tree. The posterior probability provides a natural measure of the reliability of the estimated phylogeny. Two example data sets are analyzed to infer the phylogenetic relationship of human, chimpanzee, gorilla, and orangutan. The best trees estimated by the new method are the same as those from the maximum likelihood analysis of separate topologies, but the posterior probabilities are quite different from the bootstrap proportions. The results of the method are found to be insensitive to changes in the rate parameter of the branching process. PMID- 8703099 TI - Formulating an Ecosystem Approach to Environmental Protection AB - The U.S. Environmental Protection Agency (EPA) has embraced a new strategy of environmental protection that is place-driven rather than program-driven. This new approach focuses on the protection of entire ecosystems. To develop an effective strategy of ecosystem protection, however, EPA will need to: (1) determine how to define and delineate ecosystems and (2) categorize threats to individual ecosystems and priority rank ecosystems at risk. Current definitions of ecosystem in use at EPA are inadequate for meaningful use in a management or regulatory context. A landscape-based definition that describes an ecosystem as a volumetric unit delineated by climatic and landscape features is suggested. Following this definition, ecosystems are organized hierarchically, from megaecosystems, which exist on a continental scale (e.g., Great Lakes), to small local ecosystems.Threats to ecosystems can generally be categorized as: (1) ecosystem degradation (occurs mainly through pollution) (2) ecosystem alteration (physical changes such as water diversion), and (3) ecosystem removal (e.g., conversion of wetlands or forest to urban or agricultural lands). Level of threat (i.e., how imminent), and distance from desired future condition are also important in evaluating threats to ecosystems. Category of threat, level of threat, and "distance" from desired future condition can be combined into a three dimensional ranking system for ecosystems at risk. The purpose of the proposed ranking system is to suggest a preliminary framework for agencies such as EPA to prioritize responses to ecosystems at risk.KEY WORDS: Ecosystem approach; Ecological risk assessment; Environmental protection; EPA PMID- 8703098 TI - A new transmembrane 4 superfamily molecule in the nematode, Caenorhabditis elegans. PMID- 8703100 TI - Environmental Assessment of Packaging: The Consumer Point of View AB - When marketing environmentally responsible packaged products, the producer is confronted with consumer beliefs concerning the environmental friendliness of packaging materials. When making environmentally conscious packaging decisions, these consumer beliefs should be taken into account alongside the technical guidelines. Dutch consumer perceptions of the environmental friendliness of packaged products are reported and compared with the results of a life-cycle analysis assessment. It is shown that consumers judge environmental friendliness mainly from material and returnability. Furthermore, the consumer perception of the environmental friendliness of packaging material is based on the postconsumption waste, whereas the environmental effects of production are ignored. From the consumer beliefs concerning environmental friendliness implications are deduced for packaging policy and for environmental policy.KEY WORDS: Consumer behavior; Environment; Food; Packaging; Perception; Waste PMID- 8703101 TI - Political Benefits as Barriers to Assessment of Environmental Costs in Brazil's Amazonian Development Planning: The Example of the Jatapu Dam in Roraima AB - Development projects are rapidly changing the landscape in Brazilian Amazonia. Environmental impact assessments have been required since 1986, and the regulatory system is evolving as precedents are set by each new development project. The Jatapu Dam in Roraima provides an illustration of underlying impediments to assessment of environmental costs and to due consideration being given to these assessments when decisions are made. The high priority placed on the dam by the Roraima state government is unexplainable in terms of economic returns. The place of the dam in a long-term political strategy provides the best of several possible explanations, any one of which is incompatible with a "rational" weighing of economic and environmental costs and benefits. A number of lessons can be drawn from the experience of Jatapu, but some of the problems have no solution. The barriers to rational decision making illustrated by Jatapu apply to development projects in many parts of the world.KEY WORDS: Jatapu Dam; Amazonia; Dams; Hydroelectric development; Brazil; Tropical forest; Environmental impact assessment PMID- 8703102 TI - The Cotingo Dam as a Test of Brazil's System for Evaluating Proposed Developments in Amazonia AB - The proposed Cotingo Dam in Brazil's far northern state of Roraima is examined with the objective of drawing lessons for Brazil's system of evaluating environmental, social, and financial consequences of development decisions. The Cotingo Dam illustrates the difficulty of translating into practice the principles of economic and environmental assessment. Examination of the financial arguments for the Cotingo Dam indicates that justifications in this sphere are insufficient to explain why the project is favored over other alternatives and points to political factors as the best explanation of the project's high priority. Strong pressure from political and entrepreneurial interest groups almost invariably dominates decision making in Amazonia. The analysis indicates the inherent tendency of the present system to produce decisions in favor of large construction projects at the expense of the environment and local peoples. The requirements intended to assure proper weight for these concerns, such as the report on environmental impacts (RIMA) and the public hearing, fail to serve this role. Cotingo also provides a test case for constitutional protections restricting construction of dams in indigenous lands.KEY WORDS: Hydroelectric dams; Amazonia; Indigenous peoples; Brazil; Roraima PMID- 8703103 TI - Issues, Impacts, and Implications of Shrimp Aquaculture in Thailand AB - Water quality impacts to and from intensive shrimp aquaculture in Thailand are substantial. Besides the surface and subsurface salinization of freshwaters, loadings of solids, oxygen-consuming organic matter, and nutrients to receiving waters are considerable when the cumulative impacts from water exchange during the growout cycle, pond drainage during harvesting, and illegal pond sediment disposal are taken into account. Although just beginning to be considered in Thailand, partial recirculating and integrated intensive farming systems are producing promising, if somewhat limited, results. By providing on-site treatment of the effluent from the shrimp growout ponds, there is less reliance on using outside water supplies, believed to be the source of the contamination.The explosion in the number of intensively operated shrimp farms has not only impacted the coastal zone of Thailand, but has also resulted in an unsustainable aquaculture industry. Abandonment of shrimp ponds due to either drastic, disease caused collapses or more grandual, year-to-year reductions in the productivity of the pond is common. To move Thailand towards a more sustainable aquaculture industry and coastal zone environment, integrated aquaculture management is needed. Components of integrated aquaculture management are technical and institutional. The technical components involve deployment of wastewater treatment and minimal water-use systems aimed at making aquaculture operations more hydraulically closed. Before this is possible, technical and economic feasibility studies on enhanced nitrification systems and organic solids removal by oxidation between production cycles and/or the utilization of plastic pond liners need to be conducted. The integration of semi-intensive aquaculture within mangrove areas also should be investigated since mangrove losses attributable to shrimp aquaculture are estimated to be between 16 and 32 % of the total mangrove area destroyed betweeen 1979 and 1993.Government policy needs to devote as much attention to sustainability issues as it has on promoting intensive pond culture. Such a balanced policy would include training and education monitoring and enforcement, rehabilitating abandoned ponds, managing land use within the coastal zone, more community involvement, and government reorganization to eliminate overlapping jurisdictions among agencies.As integrated aquaculture management becomes more the practice than the exception, less risk of crop failure to the industry and reduced discharge loadings from intensively managed shrimp ponds to receiving waters can be expected. Projected limitations on growing and marketing shrimp in the future, such as scarcity of land and broodstock, continued disease outbreaks, negative publicity, regulatory enforcement, water treatment and solids disposal costs, and increased competition from growers in other Asian countries will also drive the government and the industry towards adopting integrated aquaculture management.KEY WORDS: Shrimp aquaculture; Thailand; Historical practices; Environmental impacts; Sustainability; Integrated management; Water treatment; Institutional aspects PMID- 8703104 TI - Why Ecosystem Management Can't Work Without Social Science: An Example from the California Northern Spotted Owl Controversy AB - It is increasingly obvious that social science, while not a sufficient condition for making ecosystem management effective, is a necessary condition. A social science typology of ecosystems is developed, applied, and shown to have substantial and unexpected implications for the practice of ecosystem management. Ecologists and environmental scientists, in particular, will find some conclusions uncomfortable. The application involves a case material from the California northern spotted owl controversy.KEY WORDS: Ecosystem management; Social science; Adaptive management; Top-down versus bottom-up planning; Outside in versus inside-out planning PMID- 8703105 TI - Neighborhood Effects in Bird Distributions, Navarre, Spain AB - This study examines the role of neighborhood effects in the spatial distributions of selected bird species in Navarre, Spain. We employed a geographic information system (GIS) to organize the data on bird distributions and relevant environmental variables and to analyze their spatial patterns. Three bird species were selected for analysis: the European honey-buzzard (Pernis apivorus), the Eurasian hobby (Falco subbuteo), and the European pied flycatcher (Ficedula hypoleuca). Selected environmental variables of the study area were digitized to create a comprehensive data base and logistic regression models were used to evaluate the significance of each variable in the spatial distribution. The spatial patterns of bird distributions were used to extract topological relationships and to identify neighborhood effects. Although all the selected species illustrate a pattern of positive spatial autocorrelation in their distributions, the significance of neighborhood effects varies from species to species. Among the selected species, neighborhood effects are most evident in the distribution of the European pied flycatcher and are significant for the Eurasian hobby. The distribution of the European honey-buzzard is not much affected by neighborhood effects. The results suggest that examination of neighborhood effects is a prerequisite for modeling bird distributions.KEY WORDS: neighborhood effects, spatial autocorrelation, logistic regression, bird distributions. PMID- 8703106 TI - Relations of Fish Community Composition to Environmental Variables in Streams of Central Nebraska, USA AB - Nine sites on streams in the Platte River Basin in central Nebraska were sampled as part of the US Geological Survey's National Water-Quality Assessment Program during 1993-1994. A combination of canonical correspondence analysis and an index of biotic integrity determined from fish community data produced complementary evaluations of water-quality conditions. Results of the canonical correspondence analysis were useful in showing which environmental variables were significant in differentiating fish communities at the nine sites. Five environmental variables were statistically significant in the analysis. Median specific conductance of water samples collected at a site accounted for the largest amount of variability in the species data. Although the percentage of the basin as cropland was not the first variable chosen in a forward selection process, it was the most strongly correlated with the first ordination axis. A rangeland-dominated site was distinguished from all others along that axis. Median orthophosphate concentration of samples collected in the year up to the time of fish sampling was most strongly correlated with the second ordination axis. The index of biotic integrity produced results that could be interpreted in terms of the relative water quality between sites. Sites draining nearly 100% cropland had the lowest scores for two individual metrics of the index of biotic integrity that were related to species tolerance. Effective monitoring of water quality could be achieved by coupling methods that address both the ecological components of fish communities and their statistical relationships to environmental factors.KEY WORDS: Canonical correspondence analysis; Fish communities; Environmental variables; Index of biotic integrity; National Water-Quality Assessment Program; Nebraska PMID- 8703107 TI - Angler Survey Contributes to Socially Acceptable Modification of Harvest Regulations to Preserve Cutthroat Trout Fishery in Snake River, Wyoming, USA AB - This is a case study that describes a survey of anglers that was used to assist in modifying fishing regulations for indigenous trout in the Snake River, Wyoming. A mail survey of anglers who purchased 1991 Wyoming fishing licenses in the two counties adjacent to the Snake River was conducted during fall 1992. Differences in angler preferences were noted between anglers who purchased licenses in two adjacent counties with different socioeconomic structures, as well as between residents and nonresidents in each county. Anglers who purchased licenses in Teton County, where there is extensive tourism and immigration by relatively wealthy residents, tended to be more specialized and less harvest oriented. Anglers in Lincoln County, which is largely agricultural and has substantially less tourism and immigration of residents, tended to fish in many different ways and indicated more desire to harvest fish. Anglers from the two counties segregated themselves; those from Teton County primarily used the upstream portion of the study reach, and those from Lincoln County primarily used a short downstream portion of the reach. Modification of fishing regulations to reduce harvest of spawning-size cutthroat trout in the Snake River probably was acceptable to most anglers due to spatial segregation and their attitudes toward harvest.KEY WORDS: Rivers; Salmonidae; Trout; Anglers; Regulations; Wyoming PMID- 8703108 TI - Simulating Secondary Succession of Elk Forage Values in a Managed Forest Landscape, Western Washington AB - Modern timber management practices often influence forage production for elk (Cervus elaphus) on broad temporal and spatial scales in forested landscapes. We incorporated site-specific information on postharvesting forest succession and forage characteristics in a simulation model to evaluate past and future influences of forest management practices on forage values for elk in a commercially managed Douglas fir (Pseudotsuga menziesii, PSME)-western hemlock (Tsuga heterophylla, TSHE) forest in western Washington. We evaluated future effects of: (1) clear-cut logging 0, 20, and 40% of harvestable stands every five years; (2) thinning 20-year-old Douglas fir forests; and (3) reducing the harvesting cycle from 60 to 45 years. Reconstruction of historical patterns of vegetation succession indicated that forage values peaked in the 1960s and declined from the 1970s to the present, but recent values still were higher than may have existed in the unmanaged landscape in 1945. Increased forest harvesting rates had little short-term influence on forage trends because harvestable stands were scarce. Simulations of forest thinning also produced negligible benefits because thinning did not improve forage productivity appreciably at the stand level. Simulations of reduced harvesting cycles shortened the duration of declining forage values from approximately 30 to 15 years. We concluded that simulation models are useful tools for examining landscape responses of forage production to forest management strategies, but the options examined provided little potential for improving elk forages in the immediate future.KEY WORDS:Cervus elaphus; Elk; Forage; Forest management; Modeling; Secondary succession PMID- 8703109 TI - Toxic Releases from Paper Made with Recovered Wastepaper versus Virgin Wood Fiber: A Research Note AB - Toxic Release Inventory (TRI) data were used to compare average releases (kilograms per metric ton) of paper mills using primarily recovered wastepaper versus mills using primarily virgin wood fiber. Annual releases, for 79 mills, of chlorine, chlorine dioxide, chloroform, acids (hydrochloric and sulfuric), volatile organics (methyl ethyl ketone, methanol, and acetone), and ammonia were compared over the years 1987-1992. Both types of mills reported generally lower toxic releases in 1992 than in 1987; however, toxic releases in all categories were significantly lower from mills using recovered wastepaper than from mills using virgin wood fiber, strongly demonstrating that recycling has added benefits beyond reduced resource consumption. These results suggest that environmental policy should concentrate as much on increasing demand for recycled paper and developing wastepaper collection infrastructure as it does on end-of-pipe pollution abatement.KEY WORDS: Recycled paper; Toxic Release Inventory PMID- 8703110 TI - Role of Wetlands in Reducing Phosphorus Loading to Surface Water in Eight Watersheds in the Lake Champlain Basin AB - A landscape-level approach was applied to eight rural watersheds to assess the role that wetlands play in reducing phosphorus loading to surface waters in the Lake Champlain Basin. Variables summarizing various characteristics of wetlands within a watershed were calculated using a geographic information system and then compared to measured phosphorus loading through multiple regression analyses. The inclusion of a variable based on the area of riparian wetlands located along low- and medium-order streams in conjunction with the area of agricultural and nonwetland forested lands explained 88% of the variance in phosphorus loading to surface waters. The best fit model coefficients (Pload=0.86Ag+0.64For 30Ripwet+160) suggest that a hectare of riparian wetland may be many times more important in reducing phosphorus than an agricultural hectare is in producing phosphorus. These results provide additional support for the concept that protection of riparian wetlands is an important management strategy for controlling stream water quality in multiuse landscapes.KEY WORDS: Wetlands; Phosphorus; Landscape; Regression model; Riparian; Vermont PMID- 8703111 TI - Review and Comparison of Wetland Impacts and Mitigation Requirements Between New Jersey, USA, Freshwater Wetlands Protection Act and Section 404 of the Clean Water Act AB - A review of wetland impacts authorized under the New Jersey Freshwater Wetlands Protection Act (FWPA) was conducted based on permitting data compiled for the period 1 July 1988 to 31 December 1993. Data regarding the acreage of wetlands impacted, location of impacts by drainage basin and watershed, and mitigation were analyzed. Wetland impacts authorized and mitigation under New Jersey's program were evaluated and compared with Section 404 information available for New Jersey and other regions of the United States.Under the FWPA, 3003 permits were issued authorizing impacts to 234.76 ha (602.27 acres) of wetlands and waters. Compensatory mitigation requirements for impacts associated with individual permits required the creation of 69.20 ha.(171.00 acres), and restoration of 16.49 ha (40.75 acres) of wetlands. Cumulative impacts by watershed were directly related to levels of development and population growth.The FWPA has resulted in an estimated 67% reduction [44.32 ha (109.47 acres) vs 136.26 ha (336.56 acres)] in annual wetland and water impacts when compared with Section 404 data for New Jersey. For mitigation, the slight increase in wetland acreage over acreage impacted is largely consistent with Section 404 data.Based on this evaluation, the FWPA has succeeded in reducing the level of wetland impacts in New Jersey. However, despite stringent regulation of activities in and around wetlands, New Jersey continues to experience approximately 32 ha (79 acres) of unmitigated wetland impacts annually. Our results suggest that additional efforts focusing on minimizing wetland impacts and increasing wetlands creation are needed to attain a goal of no net loss of freshwater wetlands.KEY WORDS: Wetlands; Clean Water Act; Freshwater Wetland Protection Act; Nationwide permits; Mitigation; New Jersey PMID- 8703112 TI - Exploring Environmental Effects of Accidents During Marine Transport of Dangerous Goods by Use of Accident Descriptions AB - On the basis of 1776 descriptions of water transport accidents involving dangerous goods, environmental problems in connection with releases of this kind are described and discussed. It was found that most detailed descriptions of environmental consequences concerned oil accidents, although most of the consequences were described as reversible changes. It was shown that crude oil releases, on average, are approximately five times larger than releases of oil products and that oil product releases are approximately five times larger than other chemicals. Only 2% of the 1776 accidents described contained information on consequences to living organisms, and only 10% contained any information on consequences to ecosystems. A relationship was found between the minimum kilometers of shore polluted and the tonnes released in the case of shore pollution from oil accidents. Oil slicks were shown to be five times longer than broad. Gravity scales used to describe and evaluate environmental consequences were discussed.KEY WORDS: Environmental accidents; Marine transport; Dangerous goods releases; Historical accidents; Gravity scales PMID- 8703113 TI - Including Past and Present Impacts in Cumulative Impact Assessments AB - Environmental concerns such as loss of biological diversity and stratospheric ozone depletion have heightened awareness of the need to assess cumulative impacts in environmental documents. More than 20 years of experience with the National Environmental Policy Act (NEPA) have provided analysts in the United States with opportunities for developing successful techniques to assess site specific impacts of proposed actions. Methods for analyzing a proposed action's incremental contribution to cumulative impacts are generally less advanced than those for project-specific impacts.The President's Council on Environmental Quality (CEQ) defines cumulative impact to include the impacts of " past, present and reasonably foreseeable future actions" regardless of who undertakes the action. Court decisions have helped clarify the distinction between reasonably foreseeable future actions and other possible future actions. This paper seeks to clarify how past and present impacts should be included in cumulative impact analyses.The definition of cumulative impacts implies that cumulative impact analyses should include the effects of all past and present actions on a particular resource. Including past and present impacts in cumulative impact assessments increases the likelihood of identifying significant impacts. NEPA requires agencies to give more consideration to alternatives and mitigation and to provide more opportunities for public involvement for actions that would have significant impacts than for actions that would not cause or contribute to significant impacts. For an action that would contribute to significant cumulative impacts, the additional cost and effort involved in increased consideration of alternatives and mitigation and in additional public involvement may be avoided if the action can be modified so that its contributions to significant cumulative impacts are eliminated.KEY WORDS: Cumulative impacts; Environmental impact assessment; National Environmental Policy Act; Significance; Mitigation PMID- 8703114 TI - Cloning and comparative mapping of a gene from the commonly deleted region of DiGeorge and Velocardiofacial syndromes conserved in C. elegans. AB - We have identified and cloned a gene, ES2, encoding a putative 476 amino acid protein with a predicted Mr of 52,568. The gene is localized within the DiGeorge/Velocardiofacial syndrome locus on 22q11.2 and is deleted in all the patients in which a deletion within 22q11 could be demonstrated, with the exception of one patient. ES2 is expressed in all the tissues studied. Sequence comparison showed identity with five ESTs and at the amino acid level the sequence was highly similar to, and collinear with, a hypothetical C. elegans protein of unknown function. Mutation analysis was performed in 16 patients without deletion, but no mutation has been found. The cDNA sequence is conserved in mouse and is localized on MMU16B1-B3, known to contain a syntenic group in common with HSA 22q11.2. PMID- 8703116 TI - A threshold model analysis of deafness in Dalmatians. AB - To elucidate the inheritance of deafness in Dalmatian dogs, 825 dogs in 111 litters were evaluated for abnormalities in hearing through the brainstem auditory evoked response (BAER). Recorded along with their quality of hearing (normal, unilaterally deaf, or bilaterally deaf) were the sex, coat color, eye color and the presence or absence of a color patch. The analysis considered deafness an ordered categorical trait in a threshold model. The underlying, unobservable continuous variate of the threshold model was assumed to be a linear function of sex of dog, coat color (black or liver and white), color patch (presence or absence), eye color, the deafness phenotype of the parents and a random family effect. Twenty-six percent of dogs were deaf in at least one ear. Eye color, color patch, sex and the hearing status of the parents were all significant contributions to deafness. The heritability of deafness, on the continuous unobservable scale, was 0.21. This value was computed after correction for eye color, color patch, parental hearing status and sex, implying that significant genetic variation exists beyond the contribution of several single loci. PMID- 8703115 TI - Molecular cloning and structural analysis of the functional mouse genomic XPG gene. AB - The mouse XPG gene is a homolog of the human DNA excision repair gene known to be defective in the hereditary sun-sensitive disorder xeroderma pigmentosum (group G). Defects in mouse XPG have been shown to directly affect the sensitivity of cultured cells to chemotherapy agents and may play a role in tumor cell drug resistance in vivo. A full-length cosmid clone of mouse XPG was isolated by complementation of the UV sensitivity and repair defect in CHO-UV135 cells. Exon mapping determined that the gene consisted of 15 exons within 32 kb of genomic DNA. Sequencing of intron-exon boundaries revealed that mouse XPG possesses a rare class of intron previously identified in only four other eukaryotic genes; it utilizes AT and AC dinucleotides instead of the expected GT and AG within the splice junctions. Promoter analysis determined that mouse XPG is expressed constitutively and probably initiates transcription from multiple start sites, yet, unlike the yeast homolog RAD2, we found no evidence that it is UVC inducible in cultured cells. Amino acid comparison with human XPG identified a highly conserved acidic region of homology not previously described. PMID- 8703117 TI - Construction and analysis of an hn-cDNA library derived from the p-arm of pig chromosome 12. AB - Our aim is to find unidentified genes on specific pig chromosomes or chromosome fragments. Our approach has involved the construction of a heterogeneous nuclear complementary (hn-c) DNA library of the p-arm of pig Chromosome (Chr) 12, the only pig chromosome present in the pig x hamster hybrid cell line 8990. Total RNA was extracted from the cells and first-strand synthesis of hn-cDNA carried out with random and oligo dT primers. Pig hn-cDNA was isolated by amplification of first-strand synthesized hn-cDNA with primers specific for Short Interspersed Repeat Elements (SINEs) via the polymerase chain reaction (PCR). Hn-cDNAs were size selected and cloned in E. coli XL-1 blue cells with PCR-Script as the vector. The library consisted of 6000 clones. Clone inserts were amplified by PCR with vector-specific primers, and randomly picked inserts greater than 600 bp were sequenced. Homology searches were carried out with the FASTA search program on the GenEmbl database. Thirty clones were sequenced, and of these three showed strong homologies to GenEmbl sequences: (1) to sheep, mouse, human, and rat mammary gland factor (MGF); (2) to MLN-50, a gene that is amplified in human familial breast cancer and is present on human Chr 17; the latter is homologous to pig chromosome 12; (3) to a family of unassigned overlapping human ESTs. Of the other sequenced clones, seven were over 80% homologous with pig SINE sequences; three were over 75% homologous to human LINE sequences; six displayed open reading frames over a mean distance equivalent to 50 amino acids, although these showed no significant similarities with sequences in the databases. Using this approach, we have been able to identify several new genes on the p-arm of pig Chr 12. This is the first report of gene isolation from a library derived from a pig chromosome fragment. PMID- 8703118 TI - A survey of the goat genome transcribed in the lactating mammary gland. AB - To fulfill its primary function, which is to synthesize milk during the course of lactation, the mammary gland requires efficient transcriptional, translational, and secretory machineries involving multiple genes among which promising candidates underlying the genetic variation of milk production have to be found. With the aim of providing a first transcriptional profile of lactating mammary tissue, a non-normalized cDNA library has been constructed from the udder of a lactating goat. After having discarded cDNA clones encoding the major milk proteins the rapid characterization of genes expressed in this tissue, by automated partial cDNA sequencing, was used to analyze a total of 435 cDNA clones. Examination of the Expressed Sequence Tags (ESTs) for similarities with sequence databases identified 234 cDNAs corresponding to 140 unique genes or proteins. Eighty-three clones, not similar to any current database entries, representing 77 novel sequences unrelated to previously described genes, were thus identified. Tissue specificity and relative abundance of 18 of these 77 unidentified clones were examined by dot blot and RT-PCR experiments. Sequence data were subsequently used to assign six genes of unknown localization in the bovine genome, to synteny groups by use of bovine-hamster cell hybrids and PCR. PMID- 8703119 TI - Characterization of a novel human dynein-related gene that is specifically expressed in testis. AB - A novel dynein-related transcript (designated DNEL1) from human adult testis has been identified that can encode a protein with a size of 91087 Da. The complete nucleotide sequence of the open reading frame is the first to be described for a human dynein-related gene. Northern blot analysis of mRNA from 16 different tissues has shown that DNEL1 is expressed specifically in testis. Analysis of somatic cell hybrids has mapped DNEL1 to Chromosome (Chr) 17. Analysis of a panel of 129 whole genome radiaton hybrid clones including 17q22-q25.3 has placed DNEL1 in 17q distal to the ERBA2L locus. DNEL1 shares a high degree of sequence identity and amino acid similarity with the C-terminal region of the outer arm axonemal dynein beta-heavy chains derived from sea urchin and other species, but not to any gene encoding dynein intermediate or light chains described to date. The close similarity of DNEL1 to the C-terminal part of the axonemal beta-heavy chain may suggest an origin from a common progenitor gene and the testis-specific pattern of expression a possible role in sperm development or motility. PMID- 8703120 TI - Molecular cloning of a mammalian ABC transporter homologous to Drosophila white gene. AB - Pigmentation of Drosophila eyes requires the concerted action of several genes, most of which have been cloned and characterized. Three of them, white, brown, and scarlet, have been directly implicated in the import of pigment precursors into the cells. These three genes encode similar proteins, belonging to the evolutionary conserved family of ATP Binding Cassette transporters. The identification of a novel mouse gene, ABC8, closely related to white is reported here, together with an analysis of its expression profile and its comparative mapping in mouse and human genome. PMID- 8703121 TI - Sensitivity to dietary obesity linked to a locus on chromosome 15 in a CAST/Ei x C57BL/6J F2 intercross. AB - Details of a new model of diet-dependent polygenic obesity are presented. CAST/Ei (Mus m. castaneus) mice remain lean after 12 weeks on a high-fat (32 kcal% fat) diet, while C57BL/6J mice become obese. The genes responsible for the obesity segregate in an F2 population derived from an intercross between CAST/Ei and C57BL/6J mice. Quantitative trait analysis, with simple sequence length polymorphisms (SSLPs) at loci previously linked to rodent obesities, identified a quantitative trait locus (QTL) on Chromosome (Chr) 15, accounting for approximately 9% of the variance in adiposity and 14% of the variance in mesenteric depot size. This locus appears to be at the same location as the dietary obesity-3 (Do3) locus controlling body fat content, which was previously identified in an F2 population derived from an SWR/J x AKR/J cross. This is also at the same location as the multigenic obesity-4 (Mob4) locus found in BSB mice, which display spontaneous polygenic obesity. Suggestive linkage also was found at loci close to the single gene mutations Ay on Chr 2 and tub on Chr 7. PMID- 8703122 TI - Microsatellite instability in IVS3 of murine c-fes gene: tumor-associated rearrangement and mammalian divergence. AB - The murine lymphomacrophage hybrids ESb, EbF1, EbF2-c4, which express c-fes constitutively, were found by Southern analysis to bear a c-fes deletion of almost 100 bp. The deleted allele was transmitted to the metastatic hybrids by their nonexpressing, poorly metastatic T-lymphoma progenitor Eb, which also has a structurally normal c-fes allele. PCR amplification and sequencing of fes cDNA spanning exons 3-5, where the deletion mapped, ruled out any involvement of coding sequences in the rearrangement. PCR amplification of the as yet unsequenced murine c-fes IVS3 and IVS4 showed they are about 50% longer than their human and feline homologs. Sequencing of IVS4 showed no difference between tumor and control DNA. Sequencing of part of the approximately 2600-bp IVS3 was guided by the restriction analysis of PCR products from control and hybrid DNAs. This showed that differences from the control appeared to be mainly located in the 900-bp HindIII-EcoRI fragment, localized in the middle of IVS3. As all three hybrids had the same restriction map, this fragment was sequenced in one of them (ESb). A run of >200 CA repeats was found in control DNA, and a reduction in the CAn microsatellite accounted for most of the c-fes deletion in the ESb hybrid. Interestingly, the 50% reduction in the size of human and feline c-fes IVS3 as compared with the murine homolog is mostly due to contraction of the same microsatellite. PMID- 8703124 TI - D5Ucl1 and Els1 are closely linked to W on mouse chromosome 5 but lie distal to the W19H deletion. PMID- 8703123 TI - Molecular dissection of a cosmid from a gene-rich region in 17q21 and characterization of a candidate gene for alpha-N-acetylglucosaminidase with two cDNA isoforms. AB - A cosmid mapped to human Chromosome (Chr) 17q21, c140c10, was found to contain a CpG island. We completed the sequence analysis of c140c10 because of two considerations: the cosmid contained an STS from the 17-beta-hydroxysteroid dehydrogenase gene (17-HSD), which was believed to be a neighbor of the breast cancer susceptibility gene, BRCA1; CpG islands are usually associated downstream and/or upstream of human genes. Computer-based exon trapping of the cosmid sequence revealed putative additional exons. With two of those exons used as a probe to screen human placental cDNA libraries, two cDNA isoforms for a novel gene, designated as ufHSD, were isolated. The amino acid sequence of the open reading frames of the cDNA showed no significant homology to any protein in the data base. However, it is possible that our cDNAs are from the gene for alpha acetylglucosaminidase, which has recently been localized to the same region. Northern analyses show that the major isoform is expressed in all tissues tested, with the highest expression in blood leukocytes and lowest in brain. Finally, our study has shown that the 46.7-kb cosmid c140c10 encompasses loci for five genes and pseudo-genes: PsiPTP4A, ufHSD, 17-HSDI, 17-HSDII, and 22A1. PMID- 8703125 TI - Acads gene deletion in BALB/cByJ mouse strain occurred after 1981 and is not present in BALB/cByJ-fld mutant mice. PMID- 8703126 TI - Close linkage of three neuronal genes on distal mouse chromosome 15. PMID- 8703127 TI - Characterization of genes expressed early in mouse spermatogenesis, isolated from a subtractive cDNA library. PMID- 8703128 TI - Chromosomal assignment of three rat and human H-rev genes, putative tumor suppressors, down-regulated in malignantly HRAS-transformed cells. PMID- 8703129 TI - Mapping of Grn, the gene encoding the granulin/epithelin precursor (acrogranin), to mouse chromosome 11. PMID- 8703130 TI - Mapping of the Gata6 gene to mouse chromosome 18. PMID- 8703131 TI - Mapping of a murine AIDS virus-related proviral gene (Mrv6) in NOD/Lt mice to chromosome 14. PMID- 8703133 TI - The Ant1 gene maps near Klk3 on proximal mouse chromosome 8. PMID- 8703134 TI - Localization of mouse homolog of the bovine leukemia virus receptor gene on mouse chromosome 10. PMID- 8703135 TI - Chromosome mapping of the mouse and rat homologs of the human DAN gene, D1S1733E. PMID- 8703136 TI - Sublocalization of the rat brown gene (B=Tyrp1) by linkage mapping. PMID- 8703137 TI - Genetic mapping of the bovine cytochrome c oxidase subunit IV pseudogene. PMID- 8703138 TI - Human aminopeptidase N is encoded by 20 exons. PMID- 8703140 TI - Smart materials, devices, and structures. Implications for surgical practice. PMID- 8703141 TI - Laparoscopic colorectal resection. A multicenter Italian study. AB - BACKGROUND: The aim of the present study was to evaluate retrospectively the experience of six surgical units currently performing laparoscopic colorectal surgery. METHODS: From November 1991 to January 1994, 200 patients (103 male, 97 female; mean age 62.5 years) were candidates for, and received, laparoscopic colorectal resection for benign (54) or malignant (196) lesions. All the units excluded patients with locally advanced organ tumors and all cases with suspected perforation and ascites. One center submitted to laparoscopic resection only stage I and IV adenocarcinoma. All surgeons considered obesity a relative contraindication. The following data were analyzed: indications, conversion rate to open surgery, operative time, morbidity and mortality, resumption of gastrointestinal function, number of lymph nodes harvested, hospital stay. RESULTS: Twenty-one out of 200 patients were converted to open surgery (10.5%); 37 patients had a complete laparoscopic procedure (17.1%); 137 had an assisted resection (68.5%); and the remaining 5 patients had a facilitated resection. The mean operative time was 208 min (90-480) for assisted resection and 275 min (54 550) for complete laparoscopic resection. The mortality rate was 1.7%; the overall morbidity was 19.6% (major complications 11.2%). All patients quickly became ambulatory and showed a prompt resumption of gastrointestinal functions, and less postoperative pain if compared with converted cases. The average number of lymph nodes was 12.1 (range 1-32). The mean hospital stay was 8.6 days (range 5-14.5). The mean follow-up was 16 months (range 6-24). The recurrence rate 11.7%. CONCLUSIONS: Laparoscopy seems to offer the possibility of minimally invasive treatment, but long-term follow-up is needed to evaluate the efficacy of laparoscopic surgery in the treatment of colorectal cancer. PMID- 8703142 TI - Thoracoscopic surgery in the elderly population. AB - BACKGROUND: We review our experience in video-assisted thoracoscopic surgery (VATS) on patients over 75 years of age from a single institution. METHODS: From September 1992 to November 1995, 22 patients (13 males and 9 females with mean age of 78.1 years) underwent the following procedures: pleural biopsies with or without drainage and decortication (5), drainage of empyema (2), wedge lung biopsy (3), bullectomy and talc insufflation (1), major lung resections (5), mediastinal biopsy (1), pericardiectomy (3), and thymectomy (2). Pulmonary function tests of the entire group showed the mean forced expiratory volume in 1 s (FEV1) to be 1.46 1 (range 0.63-2.11) and forced vital capacity (FVC) to be 2.04 (range 0.98-3.55) 1. RESULTS: There was no mortality. Morbidity included prolonged chest drainage over 7 days in two patients (9.1%), one of whom was discharged with a drainage bag. Particularly of note was the lack of postoperative pulmonary complications. CONCLUSIONS: We conclude that VATS in the elderly population is safe. Age alone should not be a contraindication for major lung or mediastinal surgery. PMID- 8703143 TI - Emergency laparoscopy. Technical support for the laparoscopic diagnosis of intestinal ischemia. AB - BACKGROUND: Laparoscopic diagnosis of intestinal ischemia is difficult. Dark colored bowels are not a reliable indicator for infarction, because there is no correlation between color and oxygenation. The same picture is produced by intraluminal blood or feces. False diagnoses are described. METHODS: We analyzed various techniques for assessing intestinal oxygenation and perfusion to support laparoscopic diagnosis. In this study laparoscopy was performed on eight pigs. A 10-cm segment of intestine was fixed to the abdominal wall and rendered ischemic. Measurements of the ischemic segment and normal intestine were taken using laser Doppler, Doppler ultrasound, spectrophotometer, and pulse oximeter. Doppler ultrasound and pulse oximetry were unsuitable in our model, as was laser-Doppler flowmetry. RESULTS: Only the spectrophotometer proved a highly sensitive means of assessing bowel oxygenation. CONCLUSIONS: This method provides the desired additional information about intestinal oxygenation, thus helping to interpret the laparoscopic picture of dark bowels. PMID- 8703144 TI - Intraabdominal contamination after gallbladder perforation during laparoscopic cholecystectomy and its complications. AB - BACKGROUND: Gallbladder perforation often occurs during laparoscopic cholecystectomy. METHODS: The frequency and causes of gallbladder perforation as well as the relevant clinical background factors were investigated in 110 patients undergoing laparoscopic cholecystectomy. We also evaluated intraperitoneal contamination by bacteria and gallstones at the time of gallbladder perforation and investigated whether perforation caused early or late postoperative complications. RESULTS: Intraoperative gallbladder perforation occurred in 29 of the 110 patients (26.3%). It was caused by injury with an electric knife during dissection of the gallbladder bed, injury during gallbladder retraction with grasping forceps, injury during gallbladder extraction from the abdomen, and slippage of cystic duct clips (potentially causing bile and stone spillage). Perforation was more frequent in patients with positive bile cultures and in those with pigment stones (p < 0.02), but not in patients with cholecystitis or cystic duct obstruction. The peritoneal cavity was contaminated by bacteria in 11/29 patients (37.9%) and by spilled stones in 3/29 patients (10.3%). There was no difference in the incidence of postoperative complications between the patients with and without perforation either in the early postoperative period or during follow-up for 24-42 months. Only one patient developed abdominal pain and fever in the early postoperative period, and they were probably related to perforation. CONCLUSIONS: Although gallbladder perforation is sometimes unavoidable during laparoscopic cholecystectomy, the risk of severe complications appears to be minimized by early closure of perforation, retrieval of as many of the spilled stones as possible, and intraperitoneal lavage. PMID- 8703145 TI - Is male gender a risk factor for conversion of laparoscopic into open cholecystectomy? AB - BACKGROUND: Based on a clinical observation that the conversion rate of laparoscopic cholecystectomy (LC) to open cholecystectomy (OC) is higher in males, we decided to review our records and to verify whether a significant difference in conversion rates exists between sexes. METHODS: A retrospective study on conversion rates of elective laparoscopic cholecystectomy (LC) into open cholecystectomy (LC) in relation to gender was carried out in 329 patients: 267 females and 62 males. RESULTS: Our data revealed that the probability of conversion is fivefold greater in males than females, 21% vs 4.5%, respectively (p = 0.0001). We attribute this striking difference to significantly more adhesions p = 0.0002) and anatomical difficulties (p = 0.003) in males during LC, leading to conversion. CONCLUSIONS: We conclude that conversion of LC to OC is more prevalent among males and is probably attributable to a greater incidence of anatomical difficulties. PMID- 8703146 TI - Laparoscopic stapler appendectomy. A prospective study of 267 consecutive cases. AB - BACKGROUND: The value of laparoscopic appendectomy remains controversial. Therefore, we investigated the accuracy of diagnostic laparoscopy in detecting acute appendicitis and tested the applicability and safety of stapling appendectomy as a routine procedure. METHODS: Data from 267 consecutive patients with suspicion of acute appendicitis were recorded prospectively. RESULTS: Histopathological examination revealed nonperforated and perforated appendicitis in 63.3% and 13.1%, respectively, and no inflammation in 10.8%. Other pathological findings were observed in 12.7%. Diagnostic laparoscopy detected appendicitis with a sensitivity and specificity of 95.6% and 96.6%, respectively; the positive and negative predictive value were 99.5% and 74.3%, respectively. Morbidity was 10.2% in total and 40% for perforated appendicitis. Planned laparoscopic reexploration reduced morbidity by 23.4% in patients with perforated appendicitis and substantial peritonitis. Mortality was 0.4%. CONCLUSIONS: Laparoscopy improves diagnostic accuracy for acute appendicitis and laparoscopic stapling appendectomy is a safe and efficient procedure for all forms of appendicitis. PMID- 8703147 TI - Endoscopic surgery for mature teratoma of the ovary. AB - BACKGROUND: This study was undertaken to assess the value of laparoscopic surgery as treatment for benign mature teratomas of the ovary. METHODS: A total of 70 patients treated exclusively with laparoscopic surgery for ovarian mature teratoma were studied. The tumors were either enucleated with preservation of the ovary or removed by salpingo-oophorectomy. RESULTS: Ovary-preserving surgery was performed in 60 cases; salpingo-oophorectomy was accomplished in 10 cases. All tumors removed were histologically benign. No surgical complications were encountered throughout the series. In eight patients who underwent primary ovary preserving surgery, a second-look laparoscopy was undertaken with virtually no pathological findings noted. CONCLUSIONS: Our findings confirm those of other authors- when performed by experienced surgeons, laparoscopic removal of ovarian mature teratomas is a safe and recommendable alternative to laparotomy. Preoperative diagnosis and intraoperative inspection of the tumor must be as thorough as possible. PMID- 8703148 TI - Laparoscopic rectopexy for complete rectal prolapse. Clinical outcome and anorectal function tests. AB - BACKGROUND: The purpose of this study was to evaluate the clinical outcome of laparoscopic rectopexy and its effect on anorectal function investigations. METHODS: Twelve patients with complete rectal prolapse without constipation underwent laparoscopic rectopexy. Pre- and postoperative evaluation included scoring of incontinence, anorectal manometry, and anal endosonography. RESULTS: No recurrences of rectal prolapse were seen (median follow-up 19 months). Continence improved in eight of nine preoperatively incontinent patients. Two patients had mild constipation after surgery. Median maximum basal pressure measured by anorectal manometry increased from 20 to 25 mmHg (p = 0.005) and the rectoanal inhibitory reflex improved in seven patients (p = 0.03). Rectal sensitivity did not change significantly. Endosonography showed asymmetry and thickening of the internal anal sphincter and submucosa preoperatively. After surgery the maximum internal anal sphincter thickness decreased from 3.0 mm to 2.6 mm (p = 0.02). CONCLUSIONS: Laparoscopic rectopexy improved continence in our patients. Anorectal function tests show a partial recovery of the internal anal sphincter. Laparoscopic rectopexy combines the low morbidity of minimal invasive surgery with the good outcome of abdominal rectopexy. PMID- 8703149 TI - Endoscopic sphincterotomy in the treatment of postoperative biliary fistulas of hepatic hydatid disease. AB - BACKGROUND: Ten patients with postoperative external biliary fistula treated by endoscopic sphincterotomy are reported. METHODS: Nine of these patients were operated for hepatic hydatid disease and one for a liver abscess. Mean daily output of bile through the fistulae which were present for 5-39 days was approximately 500 cc. RESULTS: Treatment was successful in nine patients with closure of the fistulae in 2-15 days (mean, 7 days). No response was obtained in one patient who was reoperated, and an intrahepatic biliary duct was found to be completely eroded by the cyst wall. CONCLUSIONS: Endoscopic sphincterotomy should be the first-line treatment for postoperative external biliary fistulae related to hepatic hydatid disease. PMID- 8703150 TI - Laparoscopic right and left adrenalectomies. Surgical procedures. AB - BACKGROUND: Laparoscopic approach for adrenalectomy was recently described and the operative technique is not yet well defined. METHODS: Twenty-seven laparoscopic adrenalectomies were performed between 1992 and 1995. There were 18 women and nine men ranging in age from 31 to 70 years (mean, 50.8 years). The surgical procedure was a lateral decubitus transperitoneal flank approach in 26 patients, and a retroperitoneal approach in one. Twelve right and 15 left glands were removed. Adrenal diseases were primary aldosteronism in 20 patients, nonfunctional adenoma in four patients, Cushing adenoma in two, and an adrenal cyst in one. Median adrenal gland size was 2.0 cm (range 0.5-8 cm). RESULTS: Five patients were converted to laparotomy (18%)-for dissection problems in four and for an unrecognized gland in one. The median anesthesia time was 200 min and the median surgical time was 140 min. Operative morbidity was one adrenal vein injury sectioned close to the vena cava. The hemorrhage was controlled by laparoscopic suturing without conversion. This patient required a three-unit blood transfusion. No mortality occurred and postoperative morbidity was one minor chest infection. The median postoperative in-hospital stay was 4.6 days (range 2 8) for nonconverted patients. CONCLUSIONS: Laparoscopic adrenal gland removal is safe and offers fast recovery and short in-hospital stay. Laparoscopic adrenalectomy combines the advantages of both the conventional anterior and posterior approach. PMID- 8703151 TI - A comparison of operating room crowding between open and laparoscopic operations. AB - BACKGROUND: The clutter of equipment and lines in today's operating room (O.R.) is increasing. Endoscopic surgical procedures are particularly afflicted by this problem because they require additional equipment. Increasing O.R. crowding may present unnecessary hazards to traffic and adversely affect the performance of the surgical team. The purpose of this study is to provide a quantitative summary of the furniture, equipment, cables, and tubes present during open and laparoscopic operations. METHODS: We prospectively studied an unselected series of general surgical open (OP, n = 10) and laparoscopic (LAP, n = 10) operations performed at a major university teaching hospital. We recorded the location of all furniture and equipment as well as the source, course, and destination of all cables and tubes in the O.R. Cables and tubes touching the surgeon or the assistant were particularly noted. Results are expressed as median values for each group. RESULTS: The percent of O.R. space occupied increased from OP = 36% to LAP = 41% (p < 0.002). The median number of cables and tubes present increased from OP = 27 to LAP = 34 (p < 0.0002), with the number of these lines touching a member of the surgical team increasing from OP = 2 to LAP = 6 (p < 0.0003). CONCLUSIONS: We conclude that there is a significant trend toward increasing O.R. crowding during laparoscopic surgery. Innovative designs will be needed to reduce clutter in the O.R. of the future. PMID- 8703153 TI - Laparoscopic surgery in a small animal model. A simplified technique of retroperitoneal dissection in the rat. AB - BACKGROUND: An adequate laparoscopic small-animal model would benefit surgical oncologic research. Immunobiologic data and reagents available for the rodent make them an ideal species. We developed a simple, inexpensive, reproducible technique for laparoscopic surgery in rodents. METHODS: Carbon dioxide pneumoperitoneum is achieved in anesthetized animals. Through a 0.5-cm midline incision a 4.8-mm bronchofiberscope is inserted into the peritoneal cavity and secured with a purse-string suture (PSS). Three additional PSSs are made to introduce the dissectors. Under fiberscopic vision, a blunt dissection of the retroperitoneum exposes the inferior vena cava and aorta. Necropsy 24 h after verifies the adequacy of dissection. RESULTS: Eighteen animals survived. The only death resulted from bleeding. Mortality was 5.26%. Surgical time was 24.72 +/- 8.93 min with all animals active 2 h postlaparoscopy. CONCLUSIONS: Laparoscopic surgery (LS) can be done inexpensively without sophisticated equipment. The rodent is ideal for examining the immunologic consequences of laparoscopic surgery and pneumoperitoneum. PMID- 8703152 TI - Inferior vena caval blood flow and cardiac hemodynamics during carbon dioxide pneumoperitoneum. AB - BACKGROUND: The effects of carbon dioxide pneumoperitoneum on venous return and cardiac hemodynamics during laparoscopic surgery were studied. METHODS: Twelve adult pigs underwent placement of an electromagnetic flow meter across the infrarenal vena cava (IVC) as well as placement of Swan Ganz and arterial monitoring catheters. Measurements of the flow through infrarenal IVC, cardiac output (CO), pulmonary capillary wedge pressure (PCWP), mean arterial pressure (MAP), and heart rate were recorded at baseline, 5 and 60 min following insufflation to 15 mmHg with CO2, and 5 min following desufflation. Stroke volumes and systemic vascular resistance (SVR) were calculated as well. RESULTS: Flow through the IVC dropped by 24 and 31% at 5 and 60 min (p = 0.03 and 0.02, respectively). Paradoxically, cardiac output rose by 14 and 28% at 5 and 60 min (p = 0.03 at 60 min). Central venous and pulmonary capillary wedge pressures rose transiently by 35 and 36% at 5 min before returning to baseline (p < 0.01). Mean arterial pressure and heart rate remained relatively constant during insufflation. Systemic vascular resistance diminished from 938 dynes/cm/s prior to insufflation to its nadir at 60 min of 650 dynes/cm/s (p < 0.01). CONCLUSIONS: These observations suggest potentially complex interactions between the mechanical and systemic effects of the CO2 pneumoperitoneum on venous return. Transient elevations in cardiac filling pressures occur by an unknown mechanism, and a generalized enhanced inotropic state mediated via increased sympathetic outflow is observed in this hypercapnic anesthetized animal model. PMID- 8703154 TI - Endoscopic management of inveterate esophageal perforations and leaks. AB - The endoscopic management of four selected patients with inveterate esophageal perforations or leaks is presented. One patient had a perforation of the cervical esophagus following endoscopic removal of a foreign body already treated with surgical drainage; two patients had a leak following diverticulectomy and esophagogastrostomy, respectively, persistent after multiple surgical repairs; the last patient had a spontaneous perforation of the thoracic esophagus persistent after two transthoracic repairs. The mean time elapsed between the diagnosis of perforation and the endoscopic treatment was 19 days. In one patient, transesophageal drainage of a mediastinal abscess was performed. In the other three patients, a stent was placed to seal the leak in combination with gastric and esophageal aspiration. Two of these patients underwent endoscopy in critical condition and could have not been candidates for major surgical procedures. All patients received enteral nutrition. No morbidity or mortality related to the endoscopic procedure was recorded; the treatment was effective in all patients who recovered and resumed oral feeding within 3 weeks. We conclude that endoscopic transesophageal drainage and stenting are effective procedures in the management of patients with inveterate esophageal perforations or leaks. PMID- 8703155 TI - Laparoscopic repair of paraumbilical ventral hernia with increasing size in an obese patient. AB - We report a case of paraumbilical ventral hernia with increasing size over 1 year which was successfully repaired using laparoscopic techniques. The repair was made using a 1-mm-thick expanded polytetrafluoroethylene patch inserted intraperitoneally and then stapled to the anterior abdominal wall over the defect. This laparoscopic repair is easily accessible, is considered to reduce morbidity and is thus recommended as the first choice of treatment for ventral hernia. PMID- 8703156 TI - Laparoscopic evaluation and treatment of intestinal malrotation in infants. AB - Infants with intestinal malrotation present with bilious emesis and the diagnosis is generally obtained by an upper gastrointestinal barium study. Malrotation is suspected if the ligament of Treitz is not positioned to the left of the vertebral body. Barium enema may also be used to detect malrotation by noting the abnormal position of the cecum from its usual placement in the right lower quadrant, but this study is not as reliable due to the mobility of the cecum. Some infants may not have classic radiographic findings for malrotation, yet the contrast studies are not entirely normal. We recently treated two infants with recurrent vomiting whose UGI studies suggested intestinal malrotation. Laparoscopic exploration confirmed the diagnosis of malrotation. Laparoscopic correction (Ladd's procedure) of malrotation was carried out in one infant. The second infant underwent a traditional Ladd's procedure. The technique of laparoscopic Ladd's procedure is described. Laparoscopy may be used for the diagnosis and treatment of infants with intestinal malrotation. It may be especially helpful to verify the diagnosis in patients who do not have classic radiographic findings. Whether laparoscopy should be used in patients with midgut volvulus is debatable. Laparoscopic derotation of the volvulus in a setting where the bowel is markedly distended may be difficult and dangerous. PMID- 8703157 TI - A complication of T-fasteners in percutaneous endoscopic gastrostomy (PEG) placement. AB - Two patients with sinus tracts from retained T-fasteners following PEG tube placement are reported. Both patients had the PEG tubes subsequently removed and presented with purulent discharge and granulations near well-healed gastrostomy sites. The management of this complication and a possible method of prevention are discussed. PMID- 8703158 TI - A new reusable instrument designed for simple and secure knot tying in laparoscopic surgery. AB - This new instrument is a metal cannula designed for use in creating controlled intracorporeal knots similar to instrument ties used in open surgery. It can therefore be used in a number of situations in laparoscopic surgery for ligation in continuity-for example, of the undivided cystic duct or of a vascular pedicle or for ligation of the cut end of a pedicle held in artery forceps. The cannula is designed to be loaded with 0 suture material and has a tip that readily penetrates the abdominal wall through a 3-mm skin incision, so it can be introduced either directly or through an introducer sleeve in a laparoscopic port. A curved dissecting forceps is used for knot formation. The knot is tightened by pushing the cannula with the other hand, and further throws are applied. The tip is profiled so that the knot, which is formed is pushed down into position without slippage or damage to the suture material. PMID- 8703159 TI - A simple device for laparoscopic cholangiography. The clothes-peg clamp. PMID- 8703160 TI - Fishing-rod-type abdominal wall lifter for gasless laparoscopic surgery. AB - We have designed a new abdominal wall lifter for gasless laparoscopic surgery which consists of stainless steel rods and iron lifters. They elevate the abdominal wall up like a dome-type camping tent, which does not disturb any manipulation of scope or X-ray camera. We received a good view of the peritoneal cavity without CO2 gas insufflation in ten patients with cholecystitis. This will be helpful for general laparoscopic surgery or laparoscopic assisted surgery with the use of conventional forceps or extracorporeal suturing through a valveless trocar. PMID- 8703162 TI - Chylothorax after endoscopic thoracic sympathectomy. PMID- 8703161 TI - Pelvic abscess following laparoscopic appendectomy. PMID- 8703163 TI - Is outpatient laparoscopic cholecystectomy wise? PMID- 8703164 TI - The author replies PMID- 8703166 TI - News and notices PMID- 8703165 TI - Granting of ultrasonography privileges for surgeons. Society of American Gastrointestinal Endoscopic Surgeons. PMID- 8703167 TI - Comparison of a trial of labor with an elective second cesarean section. AB - BACKGROUND: In an attempt to reduce the rate of cesarean section, obstetricians now offer a trial of labor to pregnant women who have had a previous cesarean section. Although a trial of labor is usually successful and is relatively safe, few studies have directly addressed the maternal and perinatal morbidity and mortality associated with this method of delivery. METHODS: We performed a population-based, longitudinal study of 6138 women in Nova Scotia who had previously undergone cesarean section and had delivered a singleton live infant in the period from 1986 through 1992. RESULTS: A total of 3249 women elected a trial of labor, and 2889 women chose to undergo a second cesarean section. There were no maternal deaths. The overall rate of maternal morbidity was 8.1 percent; 1.3 percent had major complications (a need for hysterectomy, uterine rupture, or operative injury) and 6.9 percent had minor complications (puerperal fever, a need for blood transfusion, or abdominal-wound infection). Although the overall rate of maternal complications did not differ significantly between women who chose a trial of labor and the women who elected cesarean section (odds ratio for the trial-of-labor group, 0.9; 95 percent confidence interval, 0.8 to 1.1), major complications were nearly twice as likely among women undergoing a trial of labor (odds ratio, 1.8; 95 percent confidence interval, 1.1 to 3.0). Apgar scores, admission to the neonatal intensive care unit, and perinatal mortality were similar among the infants whose mothers had a trial of labor and those whose mothers underwent elective cesarean section. CONCLUSION: Among pregnant women who have had a cesarean section, major maternal complications are almost twice as likely among those whose deliveries are managed with a trial of labor as among those who undergo an elective second cesarean section. PMID- 8703168 TI - Low-molecular-weight heparin (enoxaparin) as prophylaxis against venous thromboembolism after total hip replacement. AB - BACKGROUND: The risk of venous thromboembolism in patients undergoing total hip replacement is known to be high. However, the optimal duration of prophylaxis with anticoagulant agents after this procedure is unknown. We sought to determine whether one month of anticoagulant therapy with the low-molecular-weight heparin enoxaparin is more effective than enoxaparin therapy given only during the hospitalization for surgery. METHODS: Two hundred sixty-two patients undergoing total hip replacement received enoxaparin during their hospitalizations (average stay, 10 to 11 days). They were then randomly assigned to receive enoxaparin or placebo (131 patients each). Blinded outpatient therapy (or placebo) was continued long enough that the total treatment period, inpatient plus outpatient, was one month for each patient. Bilateral ascending phlebography was performed 19 to 23 days after discharge, with deep-vein thrombosis as the primary end point. Distal and proximal thrombosis, pulmonary embolism, and hemorrhage were also recorded, as were deaths. RESULTS: Venography was adequate in 116 patients in the placebo group and 117 in the enoxaparin group. We observed 43 episodes of deep vein thrombosis and 2 episodes of pulmonary embolism in the placebo group, but only 21 episodes of deep-vein thrombosis and no episodes of pulmonary embolism in the enoxaparin group (incidence of thromboembolism, 39 percent and 18 percent, respectively; P<0.001). The difference in the incidence of proximal deep-vein thrombosis was also significant (24 percent and 7 percent in the placebo and enoxaparin groups, respectively; P<0.001). Six enoxaparin groups, respectively; P<0.001). Six patients in the enoxaparin group and one patient in the placebo group had hematomas at their injection sites. No patients died or had major complications. CONCLUSIONS: There were significantly fewer venous thromboembolic complications in patients undergoing elective hip replacement when prophylaxis with enoxaparin was given for a total of one month, rather than only during the hospitalization. PMID- 8703169 TI - A comparison of low-dose heparin with low-molecular-weight heparin as prophylaxis against venous thromboembolism after major trauma. AB - BACKGROUND: Patients who have had major trauma are at very high risk for venous thromboembolism if they do not receive thromboprophylaxis. We compared low-dose heparin and a low-molecular-weight heparin with regard to efficacy and safety in a randomized clinical trial in patients with trauma. METHODS: Consecutive adult patients admitted to a trauma center who had Injury Severity Scores of at least 9 and no intracranial bleeding were randomly assigned to heparin (5000 units) or enoxaprin (30 mg), each given subcutaneously every 12 hours in a double-blind manner, beginning within 36 hours after the injury. The primary outcome was deep vein thrombosis as assessed by contrast venography performed on or before day 14 after randomization. RESULTS: Among 344 randomized patients, 136 who received low dose heparin and 129 who received enoxaparin had venograms adequate for analysis. Sixty patients given heparin (44 percent) and 40 patients given enoxaparin (31 percent) had deep-vein thrombosis (P=0.014). The rates of proximal-vein thrombosis were 15 percent and 6 percent, respectively (P=0.012). The reductions in risk with enoxaparin as compared with heparin were 30 percent (95 percent confidence interval, 4 to 50 percent) for all deep-vein thrombosis and 58 percent (95 percent confidence interval, 12 to 87 percent) for proximal-vein thrombosis. Only six patients (1.7 percent) had major bleeding (one in the heparin group and five in the enoxaparin group, P=0.12). CONCLUSIONS: Low-molecular-weight heparin was more effective than low-dose heparin in preventing venous thromboembolism after major trauma. Both interventions were safe. PMID- 8703170 TI - Constitutively activated receptors for parathyroid hormone and parathyroid hormone-related peptide in Jansen's metaphyseal chondrodysplasia. AB - BACKGROUND: An activating mutation of the receptor for parathyroid hormone (PTH) and parathyroid hormone-related peptide (PTHrP) was recently found in a patient with Jansens's metaphyseal chondrodysplasia, a rare form of short-limbed dwarfism associated with hypercalcemia and normal or low serum concentrations of the two hormones. To investigate this and other activating mutations and to refine the classification of this unusual disorder, we analyzed genomic DNA from six additional patients with Jansen's disease. METHODS: Exons encoding the PTH-PTHrP receptor were amplified by the polymerase chain reaction (PCR), and the products were analyzed by gel electrophoresis or direct nucleotide-sequence analysis. Nucleotide changes were confirmed by restriction-enzyme digestion of genomic DNA or the PCR products. RESULTS: The previously reported mutation, which changes a histidine at position 223 to arginine (H223R), was found in genomic DNA from three of the six patients but not in DNA from their healthy relatives or 45 unrelated normal subjects. A novel missense mutation that changes a threonine in the receptor's sixth membrane-spanning region to proline (T410P) was identified in another patient but not in 62 normal subjects. In two patients with radiologic evidence of Jansen's metaphyseal chondrodysplasia but less severe hypercalcemia, no receptor mutations were detected. In COS-7 cels expressing PTH-PTHrP receptors with the T410P or H223R mutation, basal cyclic AMP accumulation was four to six times higher than in cells expressing wild-type receptors. CONCLUSIONS: The expression of constitutively active PTH-PTHrp receptors in kidney, bone, and growth-plate chondrocytes provides a plausible genetic explanation for mineral ion abnormalities and metaphyseal changes in patients with Jansen's disease. PMID- 8703172 TI - A comparison of dietary trends among racial and socioeconomic groups in the United States. AB - BACKGROUND: There may be dietary differences among racial and socioeconomic groups in the United States. METHODS: Using data from a representative sample of adults, we compared dietary trends among blacks and whites of varying socioeconomic status. We developed comparable measures of diet and of the consumption of macronutrients and food groups for 6061 participants in the 1965 Nationwide Food Consumption Surveys, 16,425 in the 1977-1978 Nationwide Food Consumption Surveys, and 9920 in the 1989-1991 Continuing Survey of Food Intake by Individuals (all conducted by the U.S. Department of Agriculture). The primary outcome was the score (range, 0 to 16) on the Diet Quality Index, a composite of eight food-and-nutrient-based recommendations from the National Academy of Sciences. A score of 4 or less was considered to indicate a relatively more healthful diet, and a value of 10 or more a relatively less healthful diet. RESULTS: Overall dietary quality improved in all groups, from a mean Diet Quality Index score of 7.4 in 1965 to 6.4 in 1989-1991. In 1965, blacks of low socioeconomic status and, to a lesser extent, whites of low socioeconomic status had better diets than whites of high socioeconomic status. By the 1989-1991 survey, the differences among racial and socioeconomic groups had narrowed. In 1965, 9.3 percent of whites of low socioeconomic status, 16.4 percent of blacks of low socioeconomic status, and 4.7 percent of whites of high socioeconomic status had mean scores of 4 or less. In the 1989-1991 survey the respective percentages were 19.9, 23.5, and 20.0. Fat consumption decreased in all groups. The consumption of fruits and vegetables varied little over time, except for an increase among blacks of medium and high socioeconomic status. The consumption of grains and legumes increased over time among whites of medium and high socioeconomic status and declined among blacks of low socioeconomic status. CONCLUSIONS: In 1965, there were large differences among groups in dietary quality, with whites of high socioeconomic status eating the least healthful diet, as measured by the index, and blacks of low socioeconomic status the most healthful. By the 1989-1991 survey, the diets of all groups had improved and were relatively similar. PMID- 8703174 TI - Phage libraries--a new route to clinically useful antibodies. PMID- 8703175 TI - Toward fewer cesarean sections--the role of a trial of labor. PMID- 8703176 TI - Actions of parathyroid hormone-related peptide and its receptors. PMID- 8703177 TI - Improving our diet--still a long way to go. PMID- 8703178 TI - The purpose of advance medical planning--autonomy for patients or limitation of care? PMID- 8703179 TI - Controlling the costs of health care for the elderly--fair means and foul. PMID- 8703181 TI - Pregnancy and the risk of stroke. AB - BACKGROUND: It is widely believed that pregnancy increases the risk of stroke, but there are few data available to quantify that risk. METHODS: We identified all female patients 15 through 44 years of age in central Maryland and Washington, D.C., who were discharged from any of 46 hospitals in the study area in 1988 or 1991. Two neurologists reviewed each case, using data from the women's medical records. We determined whether the women had been pregnant at the time of the stroke or up to six weeks before it occurred. For purposes of this analysis, the six-week period after pregnancy could begin with an induced or spontaneous abortion or with the delivery of a live or stillborn child. RESULTS: Seventeen cerebral infarctions and 14 intracerebral hemorrhages occurred in women who were or had recently been pregnant (pregnancy-related strokes), and there were 175 cerebral infarctions and 48 intracerebral hemorrhages that were not related to pregnancy. For cerebral infarction, the relative risk during pregnancy, adjusted age and race, was 0.7 (95 percent confidence interval, 0.3 to 1.6), but it increased to 8.7 for the postpartum period (after a live birth or stillbirth) (95 percent confidence interval, 4.6 to 16.7). For intracerebral hemorrhage, the adjusted relative risk was 2.5 during pregnancy (95 percent confidence interval, 1.0 to 6.4) but 28.3 for the postpartum period (95 percent confidence interval, 13.0 to 61.4). Overall, for either type of stroke during or within six weeks after pregnancy, the adjusted relative risk was 2.4 (95 percent confidence interval, 1.6 to 3.6), and the attributable, or excess, risk was 8.1 strokes per 100,000 pregnancies (95 percent confidence interval, 6.4 to 9.7). CONCLUSIONS: The risks of both cerebral infarction and intracerebral hemorrhage are increased in the six weeks after delivery but not during pregnancy itself. PMID- 8703183 TI - Intellectual impairment in children exposed to polychlorinated biphenyls in utero. AB - BACKGROUND: In utero exposure to polychlorinated biphenyls, a ubiquitous environmental contaminant, has been linked to adverse effects on neurologic and intellectual function in infants and young children. We assessed whether these effects persist through school age and examined their importance in the acquisition of reading and arithmetic skills. METHODS: We tested 212 children, recruited as newborns to overrepresent infants born to women who had eaten Lake Michigan fish contaminated with polychlorinated biphenyls. A battery of IQ and achievement tests was administered when the children were 11 years of age. Concentrations of polychlorinated biphenyls in maternal serum and milk at delivery were slightly higher than in the general population. A composite measure of prenatal exposure was derived from concentrations in umbilical-cord serum and maternal serum and milk. RESULTS: Prenatal exposure to polychlorinated biphenyls was associated with lower full-scale and verbal IQ scores after control for potential confounding variables such as socioeconomic status (P = 0.02). The strongest effects related to memory and attention. The most highly exposed children were three times as likely to have low average IQ scores (P <0.001) and twice as likely to be at least two years behind in reading comprehension (P = 0.03). Although larger quantities of polychlorinated biphenyls are transferred by breast-feeding than in utero, there were deficits only in associated with transplacental exposure, suggesting that the developing fetal brain is particularly sensitive to these compounds. CONCLUSIONS: In utero exposure to polychlorinated biphenyls in concentrations slightly higher than those in the general population can have a long-term impact on intellectual function. PMID- 8703185 TI - Effects of race and income on mortality and use of services among Medicare beneficiaries. AB - BACKGROUND: There are wide disparities between blacks and whites in the use of many Medicare services. We studied the effects of race and income on mortality and use of services. METHODS: We linked 1990 census data on median income according to ZIP Code with 1993 Medicare administrative data for 26.3 million beneficiaries 65 years of age or older (24.2 million whites and 2.1 million blacks). We calculated age-adjusted mortality rates and age- and sex-adjusted rates of various diagnoses and procedures according to race and income and computed black:white ratios. The 1993 Medicare Current Beneficiary Survey was used to validate the results and determine rates of immunization against influenza. RESULTS: For mortality, the black:white ratios were 1.19 for men and 1.16 for women (P<0.001 for both). For hospital discharges, the ratio was 1.14 (P<0.001), and for visits to physicians for ambulatory care it was 0.89 (P<0.001). For every 100 women, there were 26.0 mammograms among whites and 17.1 mammograms among blacks. As compared with mammography rates in the respective most affluent group, rates in the least affluent group were 33 percent lower among whites and 22 percent lower among blacks. The black:white rate ratio was 2.45 for bilateral orchiectomy and 3.64 for amputations of all or part of the lower limb (P<0.001 for both). For every 1000 beneficiaries, there were 515 influenza immunizations among whites and 313 among blacks. As compared with immunization rates in the respective most affluent group, rates in the least affluent group were 26 percent lower among whites and 39 percent lower among blacks. Adjusting the mortality and utilization rates for differences in income generally reduced the racial differences, but the effect was relatively small. CONCLUSIONS: Race and income have substantial effects on mortality and use of services among Medicare beneficiaries. Providing health insurance is not enough to ensure that the program is used effectively and equitably by all beneficiaries. PMID- 8703186 TI - The treatment of malaria. PMID- 8703188 TI - Partial liquid ventilation--the future is now. PMID- 8703189 TI - Race and health care--an American dilemma? PMID- 8703190 TI - The molecular regulation of the reductive pentose phosphate pathway in Proteobacteria and Cyanobacteria. AB - In phototrophic and chemoautotrophic proteobacteria, genes encoding enzymes of the Calvin-Benson-Bassham pathway of CO2 fixation are often found in clusters that are transcribed from a single promoter under control of the LysR-type transcriptional activator, CbbR. Mutations affecting CbbR prevent induction of cbb genes. Gel-retardation assays have demonstrated CbbR binding to putative regulatory regions of cbb operons, and in two cases, footprinting experiments have delimited the nucleotide sequence protected by CbbR. Fusion of cbb control sequences to reporter genes has allowed the regions required for promoter activity to be defined, and recent experiments indicate that the cbb regulon in Rhodobacter is controlled by a global two-component signal transduction system that also regulates other metabolic processes in this organism. Different ways of regulating CBB cycle enzymes that also have roles in heterotrophic metabolism have recently been discovered. In cyanobacteria, the genes of the CBB pathway are organized and regulated differently, and these oxygen-evolving phototrophic bacteria have evolved different strategies to control the assimilation of CO2. PMID- 8703191 TI - Formation of the light-harvesting complex I (B870) of anoxygenic phototrophic purple bacteria. AB - The light-harvesting (LH) complex I (B870) of anoxygenic photosynthetic purple bacteria is the oligomeric form of its subunit B820 consisting of the low molecular-weight polypeptides alpha, beta, bacteriochlorophyll (BChl), and carotenoids in the stoichiometric ratio [alpha1 beta1 (BChl2) Crt1-2]n. LHI surrounds the photochemical reaction center (RC). The major absorption band of the LHI complex is species-specific and is found at 870-890 nm; those of the subunit and the monomeric BChl a (dissolved in methanol) absorb at 820 and 770 nm, respectively. The isolated LHI complex can be reversibly dissociated to the B820 subunit or to the polypeptides and pigments by addition of detergents. Reconstitution of the B820 or the functional B870 complex is still possible after partial truncation of the N- or C-terminal regions of the alpha- or beta polypeptide or of the beta-polypeptide only. The minimal structural requirements for reconstitution of a spectrally wild-type form after truncation of the polypeptides and/or modifications of the BChl molecule are described. The insertion of the LHIalpha- and LHIbeta-polypeptides into the membrane and the in vivo assembly of LHI, studied in a cell-free system and in whole cells of Rhodobacter capsulatus, depend on the primary structures of both polypeptides, BChl, the chaperones DnaK and GroEL, membrane-bound proteins, and energized membranes. Exchanges, deletions, or insertions of amino acyl residues, especially in the conserved region of the N-terminus of the LHIalpha-polypeptide, prevent or reduce the efficiency and stability of the LHI assembly. Therefore, reconstitution of LHI in a detergent micelle does not exactly reproduce the formation of the LHI complex in the photosynthetic membrane in vivo. The N terminal domains play a crucial role in the formation of the oligomeric protein scaffold and of the pigment array. Facultatively phototrophic bacteria such as Rhodospirillum (Rsp.) rubrum or Rhodobacter (Rba.) capsulatus can adjust to changes in oxygen tension, light intensity, temperature, and substrates to grow under chemotrophic or phototrophic conditions. The photosynthetic apparatus (PSA), localized mainly on intracytoplasmic membranes (ICM), is usually synthesized only under low oxygen partial pressure. The cellular amount and composition of the PSA are modified upon changing light intensity in relation to cell growth (Drews and Golecki 1995). The morphogenesis of cellular structures like ICM is quite different from self-assembly. Self-assembly is a reversible process of aggregation of the constituents of a complex structure without protein synthesis and is driven by weak or strong forces in the interactions of the constituents. Morphogenesis results from the interplay of numerous gene products and the cellular organization and is always dependent upon pre-existent structures (Harold 1995). The morphogenesis of the photosynthetic membrane in purple bacteria has been studied in its different steps. The regulation at the transcriptional and post-transcriptional levels in purple bacteria, and the structure and morphogenesis of the ICM have been described recently (Armstrong 1995; Bauer 1995; Biel 1995; Drews and Golecki 1995; Klug 1995). In this mini review, I will focus on the minimal requirements for the in vitro assembly of light-harvesting (LH) complex I (B870) from its constituents in detergent micelles and compare the results with observations on the complex process of targeting and import of LHI polypeptides into the membrane and assembly of B870. PMID- 8703193 TI - The porins from the halophilic species Ectothiorhodospira shaposhnikovii and Ectothiorhodospira vacuolata. AB - Major outer membrane proteins with porin activity were isolated from cell envelopes of the halophilic strains Ectothiorhodospira shaposhnikovii N1 and Ectothiorhodospira vacuolata beta1. The porins were obtained as oligomers. They dissociated into monomers by heat or EDTA treatment. The molecular masses of the monomers were determined by mass spectrometry to be 39,285 and 37,160 Da for E. shaposhnikovii N1 and E. vacuolata beta1, respectively. Both were shown by analytical ultracentrifugation to be trimers of about 112, 000 Da. Circular dichroism spectra indicated predominantly beta-sheet structure. The 18 N-terminal amino acid sequences of the two porins were identical except for the amino acids in positions 12 and 14. No sequence similarity with the primary structure of known porins was found. In reconstitution experiments with lipid bilayers, the porins of E. shaposhnikovii N1 and E. vacuolata beta1 formed channels with a single-channel conductance of 1.5 and 0.7 nS, respectively, in 1 M KCl. The single-channel conductance saturated with increasing salt concentration, indicating a putative binding-site for anions in the channel since both porins exhibited anion-selectivity. For the porin of E. vacuolata beta1, but not for that of E. shaposhnikovii N1, an influence of detergent concentration on the single-channel conductance was observed. PMID- 8703194 TI - Ruminococcus hydrogenotrophicus sp. nov., a new H2/CO2-utilizing acetogenic bacterium isolated from human feces. AB - A new H2/CO2-utilizing acetogenic bacterium was isolated from the feces of a non methane-excreting human subject. The two strains S5a33 and S5a36 were strictly anaerobic, gram-positive, non-sporulating coccobacilli. The isolates grew autotrophically by metabolizing H2/CO2 to form acetate as sole metabolite and were also able to grow heterotrophically on a variety of organic compounds. The major end product of glucose and fructose fermentation was acetate; the strains also formed ethanol, lactate and, to a lesser extent, isobutyrate and isovalerate. The G+C content of DNA of strain S5a33 was 45.2 mol%. 16S rRNA gene sequencing demonstrated that the two acetogenic isolates were phylogenetically identical and represent a new subline within Clostridium cluster XIVa. Based on phenotypic and phylogenetic considerations, a new species, Ruminococcus hydrogenotrophicus, is proposed. The type strain of R. hydrogenotrophicus is S5a33 (DSM 10507). Furthermore, H2/CO2 acetogenesis appeared to be a common property of most of the species phylogenetically closely related to strain S5a33 (Clostridium coccoides, Ruminococcus hansenii, and Ruminococcus productus). PMID- 8703196 TI - Molecular analysis of the Rhodobacter capsulatus chaperonin (groESL) operon: purification and characterization of Cpn60. AB - The heat-shock protein Cpn60 (chaperonin, GroEL homologue) from the phototrophic bacterium Rhodobacter capsulatus B10 was purified to homogeneity and biochemically characterized. Native Cpn60 from R. capsulatus was shown to be a tetradecamer of 840 kDa similar to that of homologous chaperones characterized so far. Cpn60 possesses ATPase activity and promotes refolding of chaotropically denatured citrate synthase. The groESL operon of R. capsulatus was cloned using a degenerate oligonucleotide and sequenced. Two open reading frames (285 and 1,635 bp) were found; they encode Cpn10 and Cpn60, with corresponding deduced molecular masses of 10.6 and 57.6 kDa. The deduced amino acid sequences coincided perfectly with those of the amino terminus and of three tryptic peptides of purified Cpn60 from R. capsulatus. Strong evidence that R. capsulatus encodes only one copy of the groESL operon was obtained. Primer-extension analysis revealed that the groESL operon is transcribed by a -35/-10-type promoter, and that transcription was initiated from the same positions before and after heat-shock under both chemotrophic and phototrophic conditions. The major initiation site is immediately followed by the inverted repeat structure CIRCE, which has been found upstream of many bacterial heat-shock operons. A second minor transcript starts just after the CIRCE element. Although heat-shock induction of a groEL-lacZ fusion failed because of thermal inactivation of the fusion protein, Western blot analysis revealed a two- to threefold induction of cellular Cpn60 levels 45-75 min after shifting from 28 degrees C to 39 degrees C. Deletion mapping of the groESL promoter identified upstream of the promoter a 19-bp element that enhances groESL transcription eightfold and contains the AT-rich sequence dAAATTTTT, which is found at similar positions in heat-shock operons of other gram-negative bacteria. PMID- 8703197 TI - Sulfonates: novel electron acceptors in anaerobic respiration. AB - The enrichment and isolation in pure culture of a bacterium, identified as a strain of Desulfovibrio, able to release and reduce the sulfur of isethionate (2 hydroxyethanesulfonate) and other sulfonates to support anaerobic respiratory growth, is described. The sulfonate moiety was the source of sulfur that served as the terminal electron acceptor, while the carbon skeleton of isethionate functioned as an accessory electron donor for the reduction of sulfite. Cysteate (alanine-3-sulfonate) and sulfoacetaldehyde (acetaldehyde-2-sulfonate) could also be used for anaerobic respiration, but many other sulfonates could not. A survey of known sulfate-reducing bacteria revealed that some, but not all, strains tested could utilize the sulfur of some sulfonates as terminal electron acceptor. Isethionate-grown cells of Desulfovibrio strain IC1 reduced sulfonate-sulfur in preference to that of sulfate; however, sulfate-grown cells reduced sulfate sulfur in preference to that of sulfonate. PMID- 8703198 TI - The cell nucleus: an Eldorado to future calcium research? PMID- 8703199 TI - Modification of the conductance and gating properties of ryanodine receptors by suramin. AB - Suramin, a polysulfonated napthylurea, increases the open probability and the single-channel conductance of rabbit skeletal and sheep cardiac ryanodine receptor channels. The main mechanism for the increase in Po is an increase in the duration of open lifetimes. The effects on conduction and gating are completely reversible and involve an interaction with the cytosolic side of the channel. 10 mM dithiothreitol had no effect on the suramin-induced increase in conductance or Po. Therefore oxidation of sulfhydryl groups on the channels does not appear to be involved. Suramin has been used as an antagonist of ATP at P2 purinoceptors, however, we find that suramin does not antagonize the effect of ATP at skeletal or cardiac ryanodine receptor channels. The unusual gating kinetics induced by suramin suggest that it does not interact with the adenine nucleotide binding site on the ryanodine receptor but rather binds at a novel site(s). The suramin-induced changes to channel gating and conduction do not prevent the characteristic modification of single-channel properties by micromolar ryanodine. PMID- 8703200 TI - Effects of internal K+ and ABA on the voltage- and time-dependence of the outward K(+)-rectifier in Vicia guard cells. AB - One of the main effects of abscisic acid (ABA) is to induce net loss of potassium salts from guard cells enabling the stomata to close. K+ is released from the vacuole into the cytosol and then to the extracellular space. The effects of increasing cytosolic K+ on the voltage- and time-dependence of the outwardly rectifying K(+)-current (IK,out) in guard cell protoplasts (GCP) was examined in the whole-cell configuration of the patch-clamp technique. The same quantitative analysis was performed in the presence of ABA at different internal K+ concentrations ([K+]i). Varying [K+]i in the patch pipette from 100 to 270 mM increased the magnitude of IK,out in a nonlinear manner and caused a negative shift in the midpoint (V0.5) of its steady-state activation curve. External addition of ABA (10-20 microM) also increased the magnitude of IK,out at all [K+]i, but caused a shift in V0.5 of the steady-state activation curve only in those GCP loaded with 150 mM internal K+ or less. Indeed, V0.5 did not shift upon addition of ABA when the [K+]i was above 150 mM and up to 270 mM, i.e., the shift in V0.5 caused by ABA depended on the [K+]i. Both increase in [K+]i and external addition of ABA, decreased (by approximately 20%) the activation time constant (tau n) of IK,out. The small decrease in tau n, in both cases, was found to be independent of the membrane voltage. The results indicate that ABA mimics the effect of increasing cytoplasmic K+, and suggest that ABA may increase IK,out and alter V0.5 of its steady-state activation curve via an enhancement in cytosolic K+. This report describes for the first time the effects of [K+]i on the voltage- and time-dependence of IK,out in guard cells. It also provides an explanation for the quantitative (total membrane current) and qualitative (current kinetics) differences found between intact guard cells and their protoplasts. PMID- 8703201 TI - Protein-dependent reduction of the pyrene excimer formation in membranes. AB - The presence of proteins in lipid bilayers always decreases the excimer formation rate of pyrene and pyrene lipid analogues in a way that is related to the protein to-lipid ratio. Energy transfer measurements from intrinsic tryptophans to pyrene have shown (Engelke et al., 1994), that in microsomal membranes, the excimer formation rate of pyrene and pyrene fatty acids is heterogeneous within the membrane plane, because a lipid layer of reduced fluidity surrounds the microsomal proteins. This study investigates whether of not liposomes prepared from egg yolk phosphatidylcholine with incorporated gramicidin A give results comparable to those from microsomal membranes. The results indicate that the influence of proteins on the lipid bilayer cannot be described by one unique mechanism: Small proteins such as gramicidin A obviously reduce the excimer formation rate by occupying neighboring positions of the fluorescent probe and thus decrease the pyrene collision frequency homogeneously in the whole membrane plane, while larger proteins are surrounded by a lipid boundary layer of lower fluidity than the bulk lipid. The analysis of the time-resolved tryptophan fluorescence of gramicidin A incorporated liposomes reveals, that the tryptophan quenching by pyrene is stronger for tryptophans located closely below the phospholipid headgroup region because of the pyrene enrichment in this area of the lipid bilayer. PMID- 8703202 TI - Bumetanide-sensitive ion fluxes in vascular smooth muscle cells: lack of functional Na+, K+, 2 Cl- cotransport. AB - To examine the involvement of Na+,K+,2Cl- cotransport in monovalent ion fluxes in vascular smooth muscle cells (VSMC), we compared the effect of bumetanide on 86Rb, 36Cl and 22Na uptake by quiescent cultures of VSMC from rat aorta. Under basal conditions, the values of bumetanide-sensitive (BS) inward and outward 86Rb fluxes were not different. Bumetanide decreased basal 86Rb uptake by 70-75% with a Ki of approximately 0.2-0.3 microM. At concentrations ranging up to 1 microM, bumetanide did not affect 36Cl influx and reduced it by 20-30% in the range from 3 to 100 microM. In contrast to 86Rb and 36Cl influx, bumetanide did not inhibit 22Na uptake by VSMC. BS 86Rb uptake was completely abolished in Na(+)- or Cl(-) free media. In contrast to 86Rb, basal BS 36Cl influx was not affected by Nao+ and Ko+. Hyperosmotic and isosmotic shrinkage of VSMC increased 86Rb and 36Cl influx to the same extent. Shrinkage-induced increments of 86Rb and 36Cl uptake were completely abolished by bumetanide with a Ki or approximately 0.3 microM. Shrinkage did not induce BS 86Rb and 36Cl influx in (Na+ or Cl-)- and (Na+ or K+) depleted media, respectively. In the presence of an inhibitor of Na+/H+ exchange (EIPA), neither hyperosmotic nor isosmotic shrinkage activated 22Na influx. Bumetanide (1 microM) did not modify basal VSMC volume and intracellular content of sodium, potassium and chloride but abolished the regulatory volume increase in isosmotically-shrunken VSMC. These data demonstrate the absence of the functional Na+,K+,2Cl- cotransporter in VSMC and suggest that in these cells basal and shrinkage-induced BS K+ influx is mediated by (Nao+ + Clo-)-dependent K+/K+ exchange and Nao(+)-dependent K+,Cl- cotransport, respectively. PMID- 8703203 TI - Permeability and reflection coefficients of urea and small amides in the human red cell. AB - Measurement of the transport parameters that govern the passage of urea and amides across the red cell membrane leads to important questions about transport of water. It had initially been thought that small protein channels, permeable to water and small solutes, traversed the membrane (see Solomon, 1987). Recently, however, very strong evidence has been presented that the 28 kDa protein, CHIP28, found in the red cell membrane, is the locus of the water channel (see Agre et al., 1993). CHIP28 transports water very rapidly but does not transport small nonelectrolytes such as urea. The irreversible thermodynamic parameter, sigma i, the reflection coefficient, is a measure of the relationship between the permeability of the solute and that of water. If a solute permeates by dissolution in the membrane, sigma i = 1.0; if it permeates by passage through an aqueous channel, sigma i < 1.0. For urea, Goldstein and Solomon (1960) found that sigma urea = 0.62 +/- 0.03 which meant that urea crosses the red cell membrane in a water-filled channel. This result and many subsequent observations that showed that sigma urea < 1.0 are at variance with the observation that CHIP28 is impermeable to urea. In view of this problem, we have made a new series of measurements of sigma i for urea and other small solutes by a different method, which obviates many of the criticisms Macey and Karan (1993) have made of our earlier method. The new method (Chen et al., 1988), which relies upon fluorescence of the intracellular dye, fluorescein sulfonate, leads to the corrected value, sigma urea,corr = 0.64 +/- 0.03 for ghosts, in good agreement with earlier data for red cells. Thus, the conclusion on irreversible thermodynamic and other grounds that urea and water share a common channel is in disagreement with the view that CHIP28 provides the sole channel for water entrance into the cell. PMID- 8703204 TI - A bicarbonate- and weak acid-permeable chloride conductance controlled by cytosolic Ca2+ and ATP in rat submandibular acinar cells. AB - A Ca(2+)-activated Cl- conductance in rat submandibular acinar cells was identified and characterized using whole-cell patch-clamp technique. When the cells were dialyzed with Cs-glutamate-rich pipette solutions containing 2 mM ATP and 1 microM free Ca2+ and bathed in N-methyl-D-glucamine chloride (NMDG-Cl) or Choline-Cl-rich solutions, they mainly exhibited slowly activating currents. Dialysis of the cells with pipette solutions containing 300 nM or less than 1 nM free Ca2+ strongly reduced the Cl- currents, indicating the currents were Ca(2+) dependent. Relaxation analysis of the "on" currents of slowly activating currents suggested that the channels were voltage-dependent. The anion permeability sequence of the Cl- channels was: NO3- (2.00) > I- (1.85) > or = Br- (1.69) > Cl- (1.00) > bicarbonate (0.77) > or = acetate (0.70) > propionate (0.41) > > glutamate (0.09). When the ATP concentration in the pipette solutions was increased from 0 to 10 mM, the Ca(2+)-dependency of the Cl- current amplitude shifted to lower free Ca2+ concentrations by about two orders of magnitude. Cells dialyzed with a pipette solution (pCa = 6) containing ATP-gamma S (2 mM) exhibited currents of similar magnitude to those observed with the solution containing ATP (2 mM). The addition of the calmodulin inhibitors trifluoperazine (100 microM) or calmidazolium (25 microM) to the bath solution and the inclusion of KN-62 (1 microM), a specific inhibitor of calmodulin kinase, or staurosporin (10 nM), an inhibitor of protein kinase C to the pipette solution had little, if any, effect on the Ca(2+)-activated Cl- currents. This suggests that Ca2+/Calmodulin or calmodulin kinase II and protein kinase C are not involved in Ca(2+)-activated Cl- currents. The outward Cl- currents at +69 mV were inhibited by NPPB (100 microM), IAA-94 (100 microM), DIDS (0.03-1 mM), 9-AC (300 microM and 1 mM) and DPC (1 mM), whereas the inward currents at -101 mV were not. These results demonstrate the presence of a bicarbonate- and weak acid-permeable Cl- conductance controlled by cytosolic Ca2+ and ATP levels in rat submandibular acinar cells. PMID- 8703205 TI - Electrorotation of erythrocytes treated with dipicrylamine: mobile charges within the membrane show their "signature" in rotational spectra. AB - In this study, electrorotation spectra of individual cells (that is, frequency dependence of cell rotation speed) have been proved to yield information not only about the passive electric properties of cell constituents, but also about the presence of mobile charges within the plasma membrane being part of ion carrier transport systems. Experiments on human erythrocytes pretreated with the lipophilic anion dipicrylamine (DPA) gave convincing evidence that these artificial mobile charges adsorbed to the plasma membrane contributed significantly to the rotational spectrum at relatively low conductivity of the external medium (2-5 mS m-1). Theoretical integration of the mobile charge concept into the single-shell model (viewing the cell as a homogenous sphere surrounded by a membrane) led to a set of equations which predicted electrorotational behavior of DPA-treated cells in dependence on medium conductivity. The quantitative data on the partition and the transmembrane translocation rate of the DPA anion extracted from the experimental rotational spectra agreed well with the corresponding literature values. PMID- 8703213 TI - A protein phosphorylation switch at the conserved allosteric site in GP. AB - A phosphorylation-initiated mechanism of local protein refolding activates yeast glycogen phosphorylase (GP). Refolding of the phosphorylated amino-terminus was shown to create a hydrophobic cluster that wedges into the subunit interface of the enzyme to trigger activation. The phosphorylated threonine is buried in the allosteric site. The mechanism implicates glucose 6-phosphate, the allosteric inhibitor, in facilitating dephosphorylation by dislodging the buried covalent phosphate through binding competition. Thus, protein phosphorylation dephosphorylation may also be controlled through regulation of the accessibility of the phosphorylation site to kinases and phosphatases. In mammalian glycogen phosphorylase, phosphorylation occurs at a distinct locus. The corresponding allosteric site binds a ligand activator, adenosine monophosphate, which triggers activation by a mechanism analogous to that of phosphorylation in the yeast enzyme. PMID- 8703214 TI - Categorical perception of sound frequency by crickets. AB - Partitioning continuously varying stimuli into categories is a fundamental problem of perception. One solution to this problem, categorical perception, is known primarily from human speech, but also occurs in other modalities and in some mammals and birds. Categorical perception was tested in crickets by using two paradigms of human psychophysics, labeling and habituation-dishabituation. The results show that crickets divide sound frequency categorically between attractive (<16 kilohertz) and repulsive (>16 kilohertz) sounds. There is sharp discrimination between these categories but no discrimination between different frequencies of ultrasound. This demonstration of categorical perception in an invertebrate suggests that categorical perception may be a basic and widespread feature of sensory systems, from humans to invertebrates. PMID- 8703215 TI - Mechanisms of heading perception in primate visual cortex. AB - When we move forward while walking or driving, what we see appears to expand. The center or focus of this expansion tells us our direction of self-motion, or heading, as long as our eyes are still. However, if our eyes move, as when tracking a nearby object on the ground, the retinal image is disrupted and the focus is shifted away from the heading. Neurons in primate dorso-medial superior temporal area responded selectively to an expansion focus in a certain part of the visual field, and this selective region shifted during tracking eye movements in a way that compensated for the retinal focus shift. Therefore, these neurons account for the effect of eye movements on what we see as we travel forward through the world. PMID- 8703216 TI - Alpha helix-RNA major groove recognition in an HIV-1 rev peptide-RRE RNA complex. AB - The solution structure of a human immunodeficiency virus type-1 (HIV-1) Rev peptide bound to stem-loop IIB of the Rev response element (RRE) RNA was solved by nuclear magnetic resonance spectroscopy. The Rev peptide has an alpha-helical conformation and binds in the major groove of the RNA near a purine-rich internal loop. Several arginine side chains make base-specific contacts, and an asparagine residue contacts a G.A base pair. The phosphate backbone adjacent to a G.G base pair adopts an unusual structure that allows the peptide to access a widened major groove. The structure formed by the two purine-purine base pairs of the RRE creates a distinctive binding pocket that the peptide can use for specific recognition. PMID- 8703217 TI - Regulation of integrin function by the urokinase receptor. AB - Integrin function is central to inflammation, immunity, and tumor progression. The urokinase-type plasminogen activator receptor (uPAR) and integrins formed stable complexes that both inhibited native integrin adhesive function and promoted adhesion to vitronectin via a ligand binding site on uPAR. Interaction of soluble uPAR with the active conformer of integrins mimicked the inhibitory effects of membrane uPAR. Both uPAR-mediated adhesion and altered integrin function were blocked by a peptide that bound to uPAR and disrupted complexes. These data provide a paradigm for regulation of integrins in which a nonintegrin membrane receptor interacts with and modifies the function of activated integrins. PMID- 8703220 TI - Appetite-suppressing effects of urocortin, a CRF-related neuropeptide. AB - The neuropeptide corticotropin-releasing factor (CRF) is well known to act on the central nervous system in ways that mimic stress and result in decreases in exploration, increases in sympathetic activity, decreases in parasympathetic outflow, and decreases in appetitive behavior. Urocortin, a neuropeptide related to CRF, binds with high affinity to the CRF2 receptor, is more potent than CRF in suppressing appetite, but is less potent than CRF in producing anxiety-like effects and activation. Doses as low as 10 nanograms injected intracerebroventricularly were effective in decreasing food intake in food deprived and free-feeding rats. These results suggest that urocortin may be an endogenous CRF-like factor in the brain responsible for the effects of stress on appetite. PMID- 8703221 TI - The mental representation of hand movements after parietal cortex damage. AB - Recent neuroimagery findings showed that the patterns of cerebral activation during the mental rehearsal of a motor act are similar to those produced by its actual execution. This concurs with the notion that part of the distributed neural activity taking place during movement involves internal simulations, but it is not yet clear what specific contribution the different brain areas involved bring to this process. Here, patients with lesions restricted to the parietal cortex were found to be impaired selectively at predicting, through mental imagery, the time necessary to perform differentiated finger movements and visually guided pointing gestures, in comparison to normal individuals and to a patient with damage to the primary motor area. These results suggest that the parietal cortex is important for the ability to generate mental movement representations. PMID- 8703222 TI - Hypoxic-ischemic encephalopathy in areas of primary myelination: a neuroimaging and PET study. AB - The stage of regional structural and biochemical development of the central nervous system appears as a critical factor determining the distribution of hypoxic-ischemic lesions during the perinatal period. We describe the brain lesions in 12 patients who suffered hypoxia-ischemia during the perinatal period. The gestational age ranged from 35 to 42 weeks and the age at death from 2 to 16 weeks. There is one patient alive at age 18 years and a second patient at age 1 year. The cerebral cortical damage is mainly restricted to areas of primary myelination and adjacent subcortical white matter. In addition, there is thalamic, basal ganglia, brainstem, and spinal cord damage. It is postulated that selective damage occurs in those areas which at the moment of the hypoxic ischemic insult had achieved higher rates of oxygen-glucose utilization. This hypothesis is supported by studies utilizing positron emission tomography which indicates that glucose utilization in the normal human neonatal brain follows a phylogenetic order. Regions that achieved higher levels of glucose consumption are those that suffered the brunt of the damage in our term neonates. PMID- 8703223 TI - Leukotriene B4 and C4 in cerebrospinal fluid from children with meningitis and febrile seizures. AB - Concentrations of immunoreactive leukotriene C4 (LTC4) and leukotriene B4 (LTB4) in the cerebrospinal fluid from 18 patients with aseptic meningitis, including 2 patients with encephalitis and 4 patients with febrile seizures, were measured by a sensitive and specific radioimmunoassay; these results were compared with those from control subjects. The concentrations of both LTC4 and LTB4 were elevated significantly in patients with meningitis (LTC4: 115.6 +/- 47.7 pg/ml; LTB4: 1,603.0 +/- 273.5 pg/ml; n = 18) compared to controls (LTC4: 83.2 +/- 21.6 pg/ml; LTB4: 1,219.3 +/- 161.5 pg/ml; n = 12; P < .05 and P < .01, respectively). However, there was no significant increase in LT levels in patients with febrile seizures. These findings suggest that LTs may play an important role in the inflammatory response induced by viral infections of the central nervous system. PMID- 8703224 TI - Incidence of intracranial bruits in neonates. AB - Although standard texts state that intracranial bruits are common in childhood, experience suggests they are uncommon in neonates. This study investigated the incidence of intracranial bruits in 1,000 consecutive neonates. Only one bruit was heard, which was due to a transmitted cardiac murmur. Uncommon physical findings require caution in deciding whether to immediately consider it a benign finding, or to pursue additional evaluation with noninvasive imaging. We suggest that listening for intracranial bruits in neonates may be an important addition to the standard neonatal screening examination. Further study to determine if pathologic bruits can be detected in the presymptomatic state is needed. PMID- 8703225 TI - Autism in Angelman syndrome: a population-based study. AB - The aim of this study was to examine the prevalence of angelman syndrome in prepubertal school-aged children and analyze its comorbidity with autistic disorder. A clinical/psychiatric evaluation of a population-based sample of 6- to 13-year-old mentally retarded children with active epilepsy was performed. Four individuals in a total population of almost 49,000 children conformed to the clinical diagnosis of Angelman syndrome. Two of these had a typical microdeletion at chromosome 15q11-13. The minimum prevalence of Angelman syndrome was estimated at 0.008% (1: 12,000) in the examined age group. All 4 children with Angelman syndrome met full behavioral criteria for the diagnosis of autistic disorder/childhood autism. It is concluded that Angelman syndrome is uncommon, but more frequent than previously estimated. The diagnosis should be considered in all patients with combined autistic disorder, severe mental retardation, and epilepsy. The implications of the possible association of Angelman syndrome and autism are discussed. PMID- 8703226 TI - Prediction of lower developmental performances of healthy neonates by neonatal EEG-sleep measures. AB - Previous neurophysiologic studies from our laboratory have demonstrated altered EEG-sleep behavior at conceptional term ages in healthy preterm infant compared with a term cohort. Developmental assessments at 12 and 24 months of age of 16 children in each cohort were compared using MANOVA. Differences were detected on MANOVA between each cohort on Bayley mental and motor performance scores, while social skills (Vineland) and temperament (Carey) were similar. Healthy preterm infants had lower scores at 12 months of age (P < .0001) and 24 months (P < .04) than term infants before adjustment for prematurity. No group differences were observed after adjustment. For the combined cohort of 32 healthy neonates, specific neonatal EEG-sleep measures were included as predictor variables in regression analyses with Bayley mental scores as outcome variables. Lower Bayley mental scores at 12 and 24 months were associated with higher spectral EEG correlations, lower spectral EEG energies in the beta frequency ranges, fewer arousals per minute, lower rapid eye movements per minute, and shorter sleep latencies from awake state to active sleep. Significant associations were observed before adjustment for prematurity at both 12 and 24 months, and after adjustment at 12 months of age for lower spectral beta EEG energies. Lower socioeconomic class also correlated with lower developmental scores. Even in the absence of major neonatal illnesses, brain adaptation to prematurity influences later developmental outcome. Adjustment for "age equivalency" may be required up to at least 24 months of age even in a healthy preterm population. PMID- 8703228 TI - Rasmussen syndrome and long-term response to gamma globulin. AB - Rasmussen syndrome (RS) is a severe and progressive focal epilepsy of unknown etiology that leads to deterioration of motor and cognitive function. We report a 14-year-old girl who developed epilepsia partialis continua involving the left hand, mild hemiparesis, and secondarily generalized seizures. RS was confirmed by brain biopsy. The patient has been treated with intravenous gamma globulin every 4 months for 46 months. The clinical course throughout this time has been distinctly atypical for RS, with no progression in motor or cognitive deficits and rare secondarily generalized seizures. Although the mechanism for action for gamma globulin in RS is not known, an immunomodulatory role has been postulated. Evidence of an immunologically mediated process in RS and clinical experience with a growing number of patients who benefit from immunomodulatory therapy suggest that a systematic study of the efficacy of gamma globulin in comparison with other forms of medical therapy is warranted. PMID- 8703227 TI - Simultaneous bilateral pallidoansotomy for idiopathic dystonia musculorum deformans. AB - A 17-year-old Russian male with a 9-year diagnosed history of dystonia musculorum deformans manifested as severe tortipelvis, lordosis, and axial and appendicular spastic dystonia, refractory to medical therapy, is reported. This patient underwent a simultaneous bilateral pallidoansotomy with dramatic results. Postoperative evaluation revealed sustained alleviation of all dystonic symptoms and abnormal movements. Rapid recovery of useful strength in all limbs as well as dramatic improvement in coordination occurred. Bilateral posteroventral pallidotomy and pallidoansotomy in the past have proven effective in alleviation of all parkinsonian symptoms, including dyskinesia and dystonia, without the concurrent risk of intransigent side effects associated with bilateral thalamotomy or other stereotactic surgical procedures. Pallidoansotomy may prove to be the treatment of choice for idiopathic torsion dystonia and merits further investigation. PMID- 8703229 TI - Neuronal migration disorders presenting with mild clinical symptoms. AB - Two children with neuronal migration disorders and unexpectedly mild clinical symptoms are reported. The first patient was followed with the diagnosis of febrile convulsion and seizures associated with fever for 14 years. Computed tomography scans were normal. Although periodic slow waves of the left parietal cortex were detected on the first two electroencephalograms, his latest examination was normal. Magnetic resonance imaging performed at 16 years of age disclosed a left parietal schizencephaly extending between the parietal cortex and corpus callosum. The second patient was followed with the diagnosis of febrile convulsion for 2 years and later experienced afebrile seizures. On his latest visit, a posterior parietal pachygyric region and a parieto-occipital island heterotopia on the left hemisphere were diagnosed by magnetic resonance imaging. We believe that review of these patients, at the mildest end of the clinical spectrum of neural migration disorders, will contribute to a new understanding of the correlation between clinical and pathologic findings of neuronal migration disorders. PMID- 8703230 TI - Angiodysgenetic necrotizing encephalopathy or diffuse meningocerebral angiomatosis. AB - A patient with angiodysgenetic necrotizing encephalopathy or diffuse meningocerebral angiomatosis complicated by intraventricular hemorrhage, posthemorrhagic hydrocephalus, and signs of heart failure is reported. The hydrocephalus and cardiomegaly were diagnosed by fetal ultrasonography. Based on these pathologic findings, a vaginal delivery was induced at 36 weeks gestation. The patient was a female infant who died 24 hours after birth. PMID- 8703231 TI - Cerebellar dysplasia and unilateral cataract in Marinesco-Sjogren syndrome. AB - The classic features of Marinesco-Sjogren syndrome include bilateral cataracts, cerebellar ataxia, and mental deficiency with an autosomal recessive inheritance pattern. Weakness and a variety of other characteristics are present inconsistently. A limited number of neuroimaging studies have indicated that cerebellar hypoplasia is the most common finding. We report a patient with near normal intelligence, unilateral cataract, and the previously unreported magnetic resonance imaging findings of cerebellar dysplasia, arachnoid cyst, and absent septum pellucidum. A review of the literature suggests significant heterogeneity in the Marinesco-Sjogren syndrome. PMID- 8703232 TI - MR findings and neurologic manifestations in Lowe oculocerebrorenal syndrome. AB - Two patients with oculocerebrorenal syndrome are described. Both had abnormal findings on electroencephalography and developed seizure episodes. Although Patient 2 manifested abnormal electroencephalographic findings at the age of 6 years, he did not develop seizures until the age of 9 years. Phenytoin was effective for controlling seizures in both patients. On magnetic resonance examination, there were two different types of lesions. The first lesion manifested high intensity on both T2- and proton density-weighted images, suggesting gliosis or demyelination. The second lesion manifested definitely low signals on both T1- and proton density-weighted images, implying a cystic lesion. However, these lesions on magnetic resonance examination were not correlated with the severity of clinical manifestations. PMID- 8703233 TI - Dandy-Walker syndrome presenting as opisthotonus: proposed pathophysiology. AB - A patient with radiographically confirmed Dandy-Walker syndrome who presented with opisthotonus, a rarely reported clinical manifestation, is reported. From four separate pharmacologic trials (baclofen, diazepam, levodopa/carbidopa, and trihexyphenidyl), combination baclofen and diazepam therapy was determined to be most efficacious. Opisthotonus and extensor posturing remain only rudimentarily understood. We review the subject and propose a specific mechanism relating our patient's anatomic and physiologic conditions. PMID- 8703234 TI - Breath-holding spells. AB - Two particularly common, and frequently frightening, forms of syncope and anoxic seizure in early childhood are pallid and cyanotic breath-holding spells. Pallid breath-holding spells result from exuberant vagally-mediated cardiac inhibition. Cyanotic breath-holding spells are of more complex pathogenesis, involving an interplay among hyperventilation, Valsalva maneuver, expiratory apnea, and intrinsic pulmonary mechanisms. The history is the mainstay of diagnosis; videotape documentation may be possible. Performance of an electrocardiogram to evaluate for prolonged QT syndrome should be strongly considered. In patients with pallid breath-holding spells, a characteristic sequence of changes may be documented on an electroencephalogram with ocular compression, if this study is performed. Spontaneous resolution of breath-holding spell, without sequelae, is anticipated. Reassurance is the mainstay of therapy. Occasionally, pharmacologic intervention may be of benefit. PMID- 8703235 TI - Usefulness of [18F]fluorodeoxyglucose positron emission tomography in pediatric epilepsy surgery. AB - We sought to analyze our experience with pediatric epilepsy surgery patients to determine the place of [18F]fluorodeoxyglucose (FDG) positron emission tomography (FDG-PET) in the preoperative evaluation of such children relative to chronic invasive intracranial monitoring. Fifty-six children who received an interictal FDG-PET as part of a phase 1 epilepsy surgery evaluation were compared with 44 children who did not have this study in a retrospective analysis of 100 patients accrued over a 4-year period. There was no significant difference between the two groups of children in terms of age or follow-up or was there a significant difference between the FDG-PET group and the no-FDG-PET group in regard to the numbers of children who had surgery, the type of procedure done, whether chronic invasive intracranial monitoring was performed, or outcome. The hypometabolic area demonstrated on interictal FDG-PET was concordant with that of the epileptogenic zone as mapped out with ictal recordings from subdural electrodes in 2 of 13 patients in whom a complete data set was available for comparison. In the other 11 children there was either poor agreement between interictal FDG-PET and ictal electrocorticographic data or the interictal FDG-PET was normal in the face of an epileptogenic focus which was successfully mapped by invasive electrophysiologic techniques and excised. We conclude that one cannot exclude a child with intractable partial seizures from surgical consideration because the interictal FDG-PET is normal; nor is there sufficient correlation between the interictal hypometabolic area on FDG-PET and the epileptogenic zone in terms of anatomic location and size to justify forgoing chronic invasive intracranial monitoring in children with intractable partial seizures being evaluated for epilepsy surgery unless there is absolute concordance between all neuroimaging, clinical, and video-electroencephalographic data. PMID- 8703236 TI - Allergens of Bipolaris species. AB - Skin prick tests done previously revealed a significantly higher percentage of sensitization to an extract of Bipolaris sp. among atopic individuals (34/147, 23.1%) compared to non-atopic individuals. Bipolaris-specific IgE levels were quantified in sera from a representative group of 38 individuals using the Fluorescence Allergosorbent Test (FAST). Result obtained by FAST were found to be comparable to the skin prick test results (r2 = 0.60, p < 0.001 for IgE levels vs wheal sizes; r2 = 0.44, p < 0.001 for IgE levels vs erythema sizes). Characterisation of the extract's allergenic component by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) showed 28 protein bands with molecular weights (MW) ranging from 11 kDa to above 100 kDa. Immunoblotting with sera of 10 Bipolaris-sensitive (skin prick test, 3 +) individuals showed that Bipolaris spore extract contained at least 4 IgE binding proteins (MW 11-13 kDa, 16-17 kDa, 20-22 kDa and 36 kDa). All 10 sera reacted to the protein at MW 20-22 kDa, 2 sera with MW 11-13 kDa, 3 sera with 16-17 kDa and 6 sera with 36 kDa. This study has thus demonstrated that spores of Bipolaris sp. contain allergenic components which may elicit IgE-mediated reactions. PMID- 8703237 TI - Cow milk protein allergy during the first year of life: a 12 year experience at the children's hospital, Bangkok. AB - CMA should be suspected for patients aged less than one year who had persistent diarrhea and/or hematemesis with no enteric pathogen found. Confirmed diagnosis could be made by Goldman challenge test. Patients with confirmed CMA should be treated by changing the cow milk feeding to soy milk feeding. However, in our study, 17% of CMA patients were also allergic to soy protein. Thus the soy milk was replaced by the elemental formula for successful treatment of this group of patients. Beside persistent diarrhea, hematemesis, anemia and hypoalbuminemia were other possible findings among patients with CMA with or without soy protein allergy. PMID- 8703238 TI - Allergic conjunctivitis caused by sugi (Cryptomeria japonica D. Don) pollen out of season. AB - Allergic conjunctivitis caused by sugi pollen is considered to be strictly a "spring disease". However, a recent report indicated that sugi pollen is scattered not only in spring but in all seasons, especially in the autumn. We retrospectively determined the number of patients with allergic conjunctivitis caused by sugi pollen during each month for 3 consecutive years, and also investigated the growth of sugi male cones in November for four years. Diagnosis of allergic conjunctivitis was based on symptomatic complaints, clinical findings, and serum sugi pollen specific IgE measured by the Multiple Antigen Simultaneous Test 16. The annual incidence of allergic conjunctivitis peaked twice, once in spring and again in autumn. Scattering of sugi pollen occurred mostly in March and October, whilst the growth of sugi male cones was highest in November. Thus, allergy to sugi pollen can cause allergic conjunctivitis both in the spring and late autumn. PMID- 8703239 TI - A preliminary study of nasal mucociliary clearance in smokers, sinusitis and allergic rhinitis patients. AB - Mucocillary clearance of nasal mucosa was evaluated with the saccharin test. A total of 140 subjects was divided into 4 groups: smokers, allergic rhinitis patients, sinusitis patients, and a normal control group. The mean mucociliary transit time for the normal control group (n = 40) was 12 minutes, while allergic rhinitis patients (n = 40) had a mean of 14.6 minutes. The smoker group (n = 40) could be further divided into the following subcategories: (a) those who had been smoking for less than 5 years (n = 10) had a mean of 15.2 minutes, (b) those who had been smoking for more than 5 years at less than one-pack a day (n = 11) had a mean of 14 minutes, and (c) those who had been smoking for more than 5 years at more than one-pack a day (n = 19) had a mean of 16.5 minutes. For the sinusitis patients (n = 20) the mean was 16.6 minutes. The study shows that sinusitis patients, along with those smokers who had been smoking for more than 5 years at more than one-pack a day, had a mucocillary transit time that was considered to be prolonged when compared to the normal control group (p < 0.05). PMID- 8703240 TI - A comparison of admission patterns of childhood asthma in 1982 and 1992. AB - The patterns of childhood asthma admission to the Department of Pediatrics, Faculty of Medicine Siriraj Hospital, Mahidol University in 1982 and 1992 were studied and compared. The number of childhood asthma admissions were 128 cases in 1982 an 132 cases in 1992. Male to female ratio changed from 1.3:1 in 1982 to 1.9:1 in 1992. The average age on admission was 57 months in 1982 vs 62 months in 1992 which was not significantly different (p > 0.05). The admission pattern of cases under 1 year of age went down from 16% in 1982 to 6% in 1992. At emergency room, adrenaline injection was used for all asthma cases in 1982 but decreased to 30% of the patients in 1992 with the replacement of nebulized beta 2 agonist. The percentage of cases that were treated with aminophylline, corticosteroid and oxygen therapy were comparable between both periods. The most common form of steroid used in admissions changed from hydrocortisone in 1982 to methyl prednisolone in 1992. The use of antibiotics went down from 92% in 1982 to 80% in 1992. All of the admission cases in both periods recovered and were later discharged. The average period of admission was 3.9 days in 1982 and was not significant different (p > 0.05) from 4.9 days in 1992. PMID- 8703241 TI - Antitumor mechanisms of Eubacterium lentum and its components. AB - In the present study, some antitumor mechanisms of Eubacterium lentum (TYH-11) and bacterial components having antitumor effects were investigated. E.lentum induced maximum NK cell activity in C3H/He mice on day 1 after injection (90.6% against 33.9% of control at E:T ratio 50:1) and the activity was kept at a level of 48.6% on day 7. Tumoricidal peritoneal macrophages were induced 9 days after E.lentum injection into BALB/c mice (56.2% against 10.1% control at E:T ratio 10:1). Tumoricidal macrophage activity persisted at the same level for at least 11 days. Cytotoxic T lymphocyte (CTL) activity was induced only in tumor bearing mice treated with E.lentum, 4 weeks after tumor inoculation. Antitumor activity was observed in the cell wall (CW) and membrane fractions (CM) of E.lentum. CW induced NK cell activity; the activity was transient while the kinetics of NK activity by CM showed 2 peaks, on day 1 and day 7. Tumoricidal macrophages were induced by CW and the activity level was the same as that induced by whole body, while that induced by CM was at a lower level. Neither CW nor CM induced CTL in tumor bearing mice. PMID- 8703243 TI - Age-related differences on clinical and immunological manifestations of SLE. AB - The clinical and immunological manifestations of 51 children with onset of systemic lupus erythematosus (SLE) before the age of 15 were compared with those of 308 adult patients with disease onset between the age of 15-49 and another 27 elderly lupus patients whose disease onset occurred at or after the age of 50. Overall disease activity determined by mean SLEDAI score was highest in the childhood group followed by the adult and the elderly group respectively. More severe form of cutaneous involvement, adenopathy, hypertension, renal involvement with renal insufficiency and anti-nDNA antibodies occurred predominantly in the childhood lupus. The clinical features distinguishing old-age lupus were chronic disease with a long interval between the time of onset and diagnosis, higher incidence of discoid rash and lower incidence of malar rash and renal involvement. Frequencies of anti-nDNA antibodies and renal involvement gradually decreased from childhood, to adulthood and to elderly lupus respectively. Anti-Sm antibodies were predominant in the adult onset group. Genetic markers, sex hormones and senility of the immune system may play a role in these age-related differences in clinical and immunological manifestations in SLE. PMID- 8703242 TI - A comparative study of tuberculin skin test reactivity between asymptomatic HIV-1 seropositive subjects and healthy volunteers. AB - During November 1993-October 1994 tuberculin skin test reactivity (PPD-Thai Red Cross: 0.1 ml of 10 IU) was determined among 399 asymptomatic HIV-1 positive subjects and 405 healthy volunteers, 10% (40/399) had PPD-TRC induration 0-2 mm compared with 4.2% (17/405) (p = 0.001) and 43.4% (173/399) had induration > or = 10 mm compared with 53.8% (218/405) (p = 0.003) of healthy volunteers. However, the percentage of the PPD-TRC induration 5-9 mm was similar among HIV-1 seropositive subjects and healthy volunteers as 37.6% (150/399) vs 34.8% (141/405) (p = 0.4). The mean PPD-TRC reaction of HIV-seropositive subjects were 6.4 +/- 0.9 mm vs. 11.0 +/- 0.5 mm among those with CD4 lymphocyte counts 200-299 cells/mm3 compared with those > or = 300 cells/mm3 (p < 0.001). We provide support for use of induration of > or = 5 mm of PPD-TRC skin reaction for evidence of latent infection with Mycobacterium tuberculosis as the CDC recommendation in asymptomatic HIV-seropositive subjects. Consideration of tuberculosis chemoprophylaxis should have benefit, particularly in areas where M.tuberculosis is highly prevalent such as Thailand. However, among HIV-1 seropositive carriers with negative tuberculin (PPD-TRC) skin tests, there needs to be a careful evaluation and follow-up for evidence of tuberculous infection. PMID- 8703244 TI - Seroprevalence of human herpesvirus 6 and 7 infections in the Thai population. AB - Seroprevalence of human herpesvirus 6 (HHV-6) and 7 (HHV-7) was estimated in the Thai population using indirect immunofluorescence assay to determine serum antibodies to HHV-6 and HHV-7. A total of 333 serum samples obtained from umbilical cord blood and venous blood of healthy persons at Siriraj Hospital and Krabi Hospital during 1990-1993 were investigated. Of 73 infants aged 0-1 month, 73% and 78% were found tob e positive for HHV-6 and HHV-7 antibodies, respectively. Antibody to HHV-6 was detected in age groups 2-3 months (38%), 4-5 months (14%), 6 months (44%), 7-11 months (66%), 1-2 year (84%), 3-4 years (82%), 5-9 years (83%), 10-19 years (83%), 20-29 years (80%), 30-39 years (67%), and over 40 years (58%), respectively. This positive rates of HHV-7 antibody in age groups 2-3 months, 4-5 months, 6 months, 7-11 months, 1-2 years, 3-4 years, 5-9 years, 10-19 years, 21-29 years, 30-39 years, and over 40 years were 50%, 21%, 10%, 37%, 47%, 82%, 75%, 72%, 72%, 67%, and 67%, respectively. At 6 months of age as the starting time of infections, 34% (14/41) and 9% (3/41) of infants had presumed primary infections of HHV-6 and HHV-7, respectively. In the follow-up study, 53% (20/38) of children were infected with HHV-6 prior to HHV-7 and only 5% vice versa. Eighty-four percent of children had acquired antibody to HHV-6 by 1-2 years old while 82% of children had acquired antibody to HHV-7 by 3-4 years old. These results suggest that HHV-6 and HHV-7 are prevalent viruses in the Thai population. The infections of both viruses begin at 6 months of age. However, infection of HHV-7 in most children begins later. The data also provided evidence that antigenic distinction between HHV-6 and HHV-7 existed with a limited cross reactivity in an antibody test. The antibody responses to HHV-6 and HHV-7 occurred independently. PMID- 8703245 TI - Monoclonal antibody-based dot-blot ELISA for the detection of Salmonella in foods. AB - Monoclonal antibody (MAb) produced to polysaccharides in the LPS molecule of salmonellae was used in a dot-blot ELISA for detecting Salmonella in 873 food samples, ie 100 fresh chicken, 261 frozen chicken, 78 pork, 84 beef, 100 hen eggs, 100 duck eggs, 50 sea-mussels, 50 shrimps and 50 squids in comparison with the conventional culture method. Salmonella culture from foods involved the following steps: pre-enrichment, enrichment in selective medium, isolation on selective and indicator media, followed by biochemical and serological identification of appropriate colonies, respectively. The whole culture procedure took 5 days. Food samples from the selective enrichment medium were also subjected to the MAb-based dot-blot ELISA. The whole procedure of dot-blot ELISA took less than 2 hours. Among 873 food samples, salmonellae could be recovered from 7.4% of the samples by the bacterial isolation method (16% of fresh chicken, 8.8% of frozen chicken, 24.4% of pork, 3.6% of beef and 2% each of hen eggs and duck eggs, respectively). Salmonella derby were predominant among pork samples while S.paratyphi B biovar java predominated in chicken. The MAb-based dot-blot ELISA were positive in 19.5% of the food samples, i.e. 30% of fresh chicken, 27.6% of frozen chicken, 34.6% of pork, 21.4% of beef, 20% of shrimp, 16% of sea mussels, 2% of hen eggs and 4% of duck eggs. The sensitivity and specificity of the MAb-based dot-blot ELISA compared to the bacterial culture method were 81.5% and 85%, respectively. The discrepancy of the data between the culture method and the dot-blot ELISA might be due to the fact that the culture method could detect only living cells at numbers that gave at least one isolated colony on the selective/differential plate while the dot-blot ELISA detects any form of Salmonella antigen. The monoclonal antibody-based dot-blot ELISA offers several advantages over the conventional bacterial culture method when it is used for screening of Salmonella contamination in foods, especially export foods. These include rapidity, cost-effectiveness and simplicity (the dot-blot ELISA does not need highly trained personnel or equipment, in contrast to the culture method). The test can be performed in field conditions and the result can be read visually. It also offers multisample analysis at one time which renders more samples of food for screening possible, thus false negative results are fewer which, in turn, assures the quality of the export food in a cost-saving, short time frame. PMID- 8703246 TI - Hepatitis B- and hepatitis C-associated hepatocellular carcinoma: evaluation of alpha-fetoprotein as a diagnostic marker. AB - Hepatocellular carcinoma (HCC) is one of the most common cancers, especially in Asia and Africa. The prognosis of HCC is very poor because of the high malignancy and the failure of early diagnosis which is mainly dependent on the late onset of clinical symptoms. Chronic infection with hepatitis B virus (HBV) and/or hepatitis C virus (HCV) is the most commonly known risk factor for developing HCC. Mass screening and monitoring of general population or of high-risk population, by measurement of serum alpha-fetoprotein (AFP), have been implemented in several countries. However, the use of AFP as a diagnostic marker for HCC is questionable due to its limited sensitivity and specificity. This article analyzed the serum level of AFP in 72 histopathologically confirmed hepatocellular carcinoma cases in Thailand. Elevation of serum AFP was detected in 75.6%, 88.9%, 79.2% and 80.0% of patients with HBsAg, anti-HCV antibody, HBV DNA, and HCV RNA, respectively. However, only 58.8% of HCC patients without any of the four markers had elevation of serum AFP. AFP is thus not a sensitive screening marker for HCC in general population, especially in those not associated with HBV or HCV. However, since elevated serum AFP was found in most patients with evidence of HBV or HCV infection, the monitoring of serum AFP level in those high-risk patients can be valuable for screening and monitoring of hepatocellular carcinoma. PMID- 8703247 TI - The immunology of nickel-induced allergic contact dermatitis. AB - Attempts have been made to elucidate the immunopathogenesis of contact allergy; yet, the exact mechanism by which nickel-induced allergic contact dermatitis (NACD) occurs is far from clear and is discussed herein. It seems to suggest that a direct nickel-MHC class II molecule binding on the skin antigen presenting cells such as Langerhans cells (LCs) would result in Th1 cell activation. Substances such as serotonin and cytokines such as TNF-alpha produced by activated mast cells may increase adhesion molecule expression and thus, enhance T cell trafficking in the skin. Cytokines such as IFN-gamma and IL-1 and perhaps IL-12 certainly play a crucial role in the activation of Th1 cells. Along with possible function of CD8 cells, downregulation of NACD may be mediated by suppressed function of LCs via the action of activated keratinocytes-derived IL 10. Inhibition of NACD can also be generated by feeding with nickel, suggesting that the induction of oral tolerance to nickel may be beneficial for an alternative immunotherapy of nickel allergy. Nevertheless, this testable model provides a direction for further investigation. PMID- 8703248 TI - Effect of inhaled corticosteroids on bronchial hyperresponsiveness in patients with mild asthma. AB - We studied the effect of inhaled budesonide on bronchial hyperresponsiveness (BHR) in twenty mild asthmatic patients. The study was conducted as a randomized, double-blind, placebo-controlled study. Before entering the study, the patients performed methacholine inhalation challenge (MIC) using a reservoir method to assess BHR. Then, they were randomly allocated to receive budesonide turbuhaler (200 micrograms/dose) or placebo turbuhaler two inhalations, twice daily for eight weeks. During the study, each patient recorded daily asthma score and daily number of puffs of beta 2 agonist and they were assessed at weeks 4 and 8. At the end of the treatment, MIC was repeated again. Patients receiving budesonide showed a significant improvement in airway responsiveness compared with those receiving placebo (p < 0.05). They also showed a significant improvement in asthma severity score and a significant decrease in beta 2 agonist bronchodilator use. This study also suggested that inhaled corticosteroids may be the primary treatment in patients, even with mild asthmatic and well-controlled symptoms. PMID- 8703249 TI - The modulatory effect of antigen- and PAF-induced asthmatic reaction by aerosol administration of OKY-046 in guinea pigs. AB - The therapeutic effect of a thromboxane A2 (TXA2) synthetase inhibitor on asthma is still controversial. This study was aimed at clarifying its effect on asthmatic reactions in guinea pigs. Both ovalbumin (OVA)- and platelet activating factor (PAF)-induced dual phase airway spasm and hyperreactivity in guinea pigs were used as the asthma model. Our results demonstrated that aerosol administration of OKY-046 could inhibit both OVA- and PAF-induced late phase bronchoconstriction and airway hyperreactivity to methacholine in OVA sensitized guinea pigs. PAF administration could also induced dual phase bronchoconstriction in normal guinea pigs. Similarly, late phase airway spasm and airway hyperreactivity after PAF exposure was also blocked by OKY-046. In conclusion, aerosol administration of OKY-046 is a safe and effective way to modulate OVA- and PAF-induced asthmatic reactions. The protective effect of OKY-046 on OVA- and PAF-induced late phase bronchoconstriction and airway hyperreactivity indicates that TXA2 might play an important role in the late phase asthmatic reaction and airway hyperreactivity. The normalization of PAF-induced airway hyperreactivity by OKY-046 also indicates that PAF induced airway inflammation might be through the generation of TXA2. PMID- 8703250 TI - Bee venom hypersensitivity and its management: patients perception of venom desensitisation. AB - The objectives of the study were to review bee venom immunotherapy from the patient's perspective: in particular its benefits and its problems, and to investigate any genetic tendency for bee venom hypersensitivity. A self administered, 9 item questionnaire was sent to 219 patients who had undergone either inpatient or outpatient bee venom immunotherapy at Flinders Medical Center. The clinic records of these patients were also reviewed. The controls for the genetic study were sought from patients, staff and students at Flinders University and Flinders Medical Centre. One hundred and forty-six questionnaires (some incomplete and anonymous) were received. The female to male ratio was 1:2.5. The age at the time of the initial anaphylactic reaction to a bee sting ranged between 2 to 59 years, with 67% of patients being less then 20 years old. Forty percent of patients underwent venom immunotherapy for a period less than 2 years with only 11% maintaining therapy for the recommended period of 5 years or more. Thirty three percent of patients stopped their therapy on their own accord. Bee stings occurring during bee venom immunotherapy (n = 56) were generally well tolerated except in 8 subjects, 7 of whom had not reached the maintenance dose. The reduction in systemic reactions to subsequent bee stings was significantly better in the study group receiving bee venom than in an historic control group treated with whole bee extract (p = 0.03). Fear of bee stings and restricted life styles were improved during or after venom immunotherapy. The frequency of a positive family history of systemic reactions to bee stings in the patient cohort was 31%, whereas in controls it was 15% (p = 0.013). Bee venom immunotherapy has dual benefits: patients are protected from subsequent sting anaphylaxis and there is reduced psychological morbidity. However, to be effective, venom immunotherapy requires a prolonged period of carefully supervised treatment and each venom injection can cause local and systemic side effects. Genetic factors appear to be present in those patients who develop immediate hypersensitivity to be stings. PMID- 8703251 TI - Supportive care by maternity nurses: a work sampling study in an intrapartum unit. AB - BACKGROUND: This work sampling study examined how much time intrapartum unit nurses spend providing supportive care overall and during weekday and weekend shifts, and by patient and staff characteristics at a university hospital with 4000 births per year in Montreal, Quebec. METHODS: Four-hour observation periods were randomly selected to represent each shift and day of the week. Within each period, eight 15-minute observation times were randomly selected. Observers located each nurse assigned to the unit at that time and recorded her activity. Supportive activities included physical comfort, emotional support, instruction, and advocacy. RESULTS: The percentage of time spent in supportive care was 6.1 percent (95% confidence interval 5.3%, 6.9%), based on 3367 observations. The time providing supportive care was similar for weekday and weekend shifts. Nurses with less than seven years of intrapartum experience spent 2.7 percent (0.9, 4.5) more time providing supportive care than nurses with seven years of experience or more. Supportive care was 9.2 percent (0.7, 17.7) greater for nulliparous than for parous women, and supportive care of women with epidural anesthesia was similar to those without it. CONCLUSION: We concluded that intrapartum unit nurses spent a small amount of time providing supportive care to women in labor. This suggests the need for perinatal caregivers and hospital administrators to reexamine how nurses spend their time, given the evidence from randomized trials showing the beneficial effects of continuous support on labor and birth outcomes. PMID- 8703252 TI - Postterm pregnancy: putting the merits of a policy of induction of labor into perspective. AB - BACKGROUND: Several randomized, controlled trials compared the policies of induction of labor and expectant management for women who reach 41 weeks' gestation, and although they suggest a better outcome for mothers and infants with such a policy in place, controversy continues as to which is the better form of care. The Canadian Multicenter Postterm Pregnancy Trial (CMPPT) enrolled 3407 women, of whom 1701 were randomized to a policy of induction of labor (induced group) and 1706 were randomized to a policy of expectant management (expectant group). Secondary analyses of data from the CMPPT were undertaken to explore a number of controversial issues. METHODS: We used data from the CMPPT to explore further the timing of delivery for women enrolled between 41 0/7 and 41 6/7 weeks' gestation, the potential impact of more liberal use of prostaglandins on cesarean section rates, and the relative merits of induced versus spontaneous labor in the two groups. RESULTS: Most women in the CMPPT (89%) were enrolled at 41 0/7 to 41 6/7 weeks' gestation, of whom 86.2 percent in the induced group and 63.6 percent in the expectant group gave birth before 42 weeks' gestation. Assuming that administration of prostaglandins would reduce the likelihood of cesarean section by 12 to 15 percent, cesarean section rates were reduced in the induction group from 21.2 percent to 20.8 to 20.9 percent, and in the expectant group from 24.5 percent 23.3 to 24.2 percent. If labor was induced as part of a policy of expectant management, the cesarean section rate was much higher (33.5%) than if labor was either spontaneous or induced as part of a policy of induction (18.5%, 22.4%). CONCLUSIONS: Women should be informed of the benefits and risks associated with the policies of induction of labor and expectant management, and their preferences regarding these policies should be respected. PMID- 8703253 TI - Why is the cesarean delivery rate so high in Alabama? An examination of risk factors, 1991-1993. AB - BACKGROUND: Cesarean delivery is now the most frequently performed major operative procedure for childbearing women in the United States. Many of these operations are reported to be unnecessary, and millions of dollars could be saved by reducing their frequency. METHODS: Method of delivery was added to the 1989 revision of the standard certificate of live birth. Alabama also added questions on the source of payment for delivery and the provider of prenatal care in 1991, which enabled an investigation of the risk factors for cesarean delivery that occurred in the state during this period. RESULTS: One of every four births in Alabama is by cesarean delivery. The risk of cesarean delivery is not random, and the risk factors include mother's race and age, coverage by private insurance, birthweight, setting where the mother received prenatal care, mother's educational attainment, live birth order, and complications of labor and delivery. CONCLUSION: With present concerns about health care reform and the costs of health care, a reduction in the cesarean delivery rate could result in significant cost savings. PMID- 8703254 TI - Early versus late prenatal care in New Mexico: barriers and motivators. AB - BACKGROUND: Adequate prenatal care is important for ensuring a good outcome for pregnant women and their children, and its initiation in the first trimester is a major component. We investigated barriers and motivators for women receiving early versus late prenatal care. METHODS: A self-administered questionnaire was distributed to 205 women who began prenatal care at the University of New Mexico Hospital Obstetrics Clinic in Albuquerque, New Mexico. The questionnaire assessed demographic information, insurance status, feelings about the pregnancy, health behaviors, and barriers and motivators to initiating prenatal care. The participants were Hispanic (41%) and non-Hispanic white (46%), low-income women with Medicaid (42%) or no health insurance (41%). They were divided into early (1st trimester 67%) and late (after 1st trimester 33%) care groups. RESULTS: Ethnicity, education, income, and age did not predict initiation of prenatal care. Late initiators cited financial problems (26%), not being aware of the pregnancy (15%), and dislike of going to doctors (14%) as reasons for the delay in seeking prenatal care. Over two-thirds of the pregnancies were unplanned, but 82 percent of the women felt positive about the pregnancy. CONCLUSIONS: Our study documents the continuing need for public health efforts to encourage women to seek early prenatal care. Specific attention should be directed toward women's perceived reasons for not initiating early care. PMID- 8703255 TI - Women's satisfaction with prenatal care settings: a focus group study. AB - BACKGROUND: Patient satisfaction is considered, together with health status, to be an outcome of the delivery of health care services as well as a measure of its quality. A focus group study of 50 low-income Mexican-American Puerto Rican, African-American, and white women in Chicago, Illinois, explored the characteristics of prenatal care that affect women's satisfaction. METHODS: Transcripts from the focus groups were analyzed using researcher-derived coding categories to develop broad themes. RESULTS: Despite their diverse ethnic backgrounds, participants revealed few differences with respect to what they value in prenatal care. Aspects of care that appeared to affect women's satisfaction included the "art of care, " the technical competence of the practitioner, continuity of caregiver, and the atmosphere and physical environment of the care setting. The one characteristic that did not appear to affect satisfaction was the caregiver's ethnicity. CONCLUSION: Knowledge of how the characteristics of prenatal care affect women's satisfaction can help increase use of care and ultimately improve perinatal outcomes. PMID- 8703256 TI - Whither tocolysis? PMID- 8703257 TI - The role of tocolysis in the prevention of preterm birth. PMID- 8703258 TI - Tocolytics--more good than harm, or is it the reverse? PMID- 8703259 TI - Tocolytics: the neonatal perspective. PMID- 8703260 TI - Sheila Kitzinger's letter from Europe: birth speak. PMID- 8703261 TI - The follow-up of a tiny baby. PMID- 8703262 TI - The primary purpose of childbirth education programs is to inform parents. PMID- 8703263 TI - Predicting the duration of lactation: evidence from a national survey. AB - BACKGROUND: Despite caregiver and policy support for breastfeeding, rates for initiating and duration of breastfeeding fell far short of Healthy People 2000 goals during the 1980s. METHODS: Data from the 1988 National Maternal-Infant Health Survey, collected from January 1989 through June 1991, were analyzed to examine predictors of duration of lactation for a sample of 2372 breastfeeding women. We conducted comparisons between women who fully breastfed and those who partially breastfed using logistic regression analysis. RESULTS: Mothers were more likely to breastfeed for longer than six months if they fully breastfed during the first month postpartum, were nonsmokers, were of higher parity, were consistent in their prenatal intent to breastfeed fully or partially and in their postpartum behaviors, participated in childbirth education classes, and delayed their return to work postpartum. CONCLUSIONS: In this study sample, although rates did not meet Healthy People 2000 goals for duration of breastfeeding, some predictors of duration were identified that can be affected by programmatic support or public policy. Our findings indicated that variables that are associated with breastfeeding and longer duration of the practice are typically correlated with social status. To support the development of breastfeeding as the cultural norm, interventions targeting breastfeeding outcomes should consider social status, ethnicity, and cultural factors. PMID- 8703264 TI - [Changes in patterns of medication as function of age of the aging patient (review of the literature)]. PMID- 8703265 TI - Hypertension secondary to adrenal tumors ten years experience. AB - We have presented our ten years experience in the surgical management of adrenal tumors. Patients presenting with hypertension (9) are normotensive following the removal of the offending adrenal tumor. One patient had an extra-adrenal pheochromocytoma, one had a huge organized adrenal hematoma and one patient has a non-functional tumor. The diagnosis and management of endocrine tumors is a challenge to the clinician. PMID- 8703266 TI - Surgical management of hyperparathyroidism. A 24 years experience. AB - Our 24 years experience in the surgical treatment of 130 patients with primary hyperparathyroidism was presented. A single adenoma of the parathyroid glands was the predominant lesion. The lower parathyroid glands were more frequently the site of the adenoma, with the right lower gland presenting the highest incidence. The incidence of hyperplasia and carcinoma is similar to the reported literature. The offending glands were identified in the vast majority of the patients. The number of complications is significantly low with a zero surgical mortality. The utilization of preoperative localizing test is not recommended for the initial neck exploration. Satisfactory results can be obtained with a minimal work up. PMID- 8703267 TI - [Rebirth of perineal prostatectomy in the era of laparoscopic lymphadenectomy]. AB - Radical prostatectomy has become the gold standard treatment for localized prostate cancer. With the introduction of Laparoscopy Lymphadenectomy for Staging of Prostate Cancer in 1991, interest in the perineal approach to radical prostatectomy has been renewed. Historically, the perineal approach has proven to be less morbid than the retropubic counterpart. Herein, the authors report their experience with 98 male patients with clinically localized T2B (or less) Cancer of the Prostate who underwent Laparoscopic Lymphadenectomy of which 83, subsequently, had a radical perineal prostatectomy. The mean follow up of this group of patients was 16.5 months. Clinicopathological results are discussed and compared with the results of radical retropubic prostatectomy in the literature. IN CONCLUSION: (1) laparoscopic lymphadenectomy with radical perineal prostatectomy are less morbid, with a faster recovery rate than the standard radical retropubic prostatectomy approach. (2) as better non-surgical methods evolve in predicting the status of the pelvic lymph nodes, reserving laparoscopic lymphadenectomy for patients with high risk of developing lymph nodes' metastasis, the perineal prostatectomy and laparoscopic lymph node dissection are becoming the state of the art treatment in patients with localized prostate cancer. PMID- 8703268 TI - Radiologic manifestations of hyperparathyroidism. PMID- 8703269 TI - Bacillary angiomatosis: microbiology, histopathology, clinical presentation, diagnosis and management. AB - Bacillary angiomatosis is known to be caused by a rickettsial organism; Rochalimaea henselae. This causative agent has been compared with different microorganisms and clinical conditions that appear in similar settings buy have been clearly differentiated from them; e.i. Cat-scratch disease (Afipia felis), Bartonella bacilliformis, other Rochalimaea sp., Kaposi's sarcoma, Lobular capillary hemangioma, Angiosarcoma, and Epithelioid hemangioma. Clinically the bacillary angiomatosis (BA) skin lesions vary from a single lesion to thousands. The cutaneous lesion appears as a bright-red round papule, subcutaneous nodule, or as a cellulitic plaque. When the lesion is biopsied it tends to blanch-out, bleed, and cause pain. The patient might present with signs and symptoms of chills, headaches, fever, malaise, and anorexia with or without weight loss. The extracutaneous lesions found in BA tend to be from multiple organs affecting from the oral lesions to anal mucosal lesions to widespread visceral lesions. The sites of preference for BA lesion manifestation tend to be the liver, spleen, lymph nodes, and bone. To diagnose bacillary angiomatosis the physician should prepare a differential diagnosis based primarily on its histopathological and clinical characteristics. To confirm the results from the stain, electron microscopy can identify the bacillus and pin-point the diagnosis of bacillary angiomatosis. The lesions presented by BA respond well to therapy with erythromycin 500 mg four times daily for a duration of 2 weeks to 2 months. In case of intolerance to erythromycin the second line of drug that successfully treats the BA bacillus is doxycyline. If relapses of the BA lesion recur, then a prolonged antibiotic therapy is necessary and in AIDS patients the duration may be extended as life-long suppressive therapy. PMID- 8703270 TI - Biliary atresia: an overview. AB - Biliary Atresia is the most common cause of persistently direct (conjugated) hyperbilirubinemia in the first three months of life. It is a progressive inflammatory obliteration of the extra- and intrahepatic bile ducts whose pathogenesis remains speculative. This review updates the history, etio pathological considerations, clinical manifestations, methods of diagnosis, surgical management, and results of this terrible disease in young infants. We will emphasize that persistent jaundice in the newborn period must be managed urgently, a diagnosis should be established early in life, and Kasai portoenterostomy offered to those infants with Biliary Atresia before their eighth week of life. This will allow more than one-third of children to survive. Hepatic transplantation is reserved for those children with failed portoenterostomy, progressive liver failure or late-referral to surgery. PMID- 8703271 TI - An evaluation of the New Zealand graduated driver licensing system. AB - Young drivers have a disproportionately high risk of experiencing a road traffic crash. On 1 August 1987 a Graduated Driver's Licensing System (GDLS) was introduced in New Zealand. This system was designed to give young drivers (i.e. 15-24 years inclusive) experience in driving while being excluded from high risk driving situations. We sought to determine the impact of the GDLS on serious injury crashes. The source of the injury crash data was New Zealand's Health Information Services' national public hospital inpatient morbidity data files for the years 1979-1992 inclusive. We disaggregated the occupant data into three age groups 15-19 years, 20-24 years, and persons 25 years of age or older and compared their trends in injury. In order to determine whether the incidence of motor vehicle crashes was simply following trends in other injury events we also included two 15-19 year old non-traffic injury comparison groups. Using time series analyses we showed that the introduction of the GDLS was closely followed by substantial reductions in car crash injuries for all age groups, especially 15 19 year olds (23% reduction). After considering effects for older occupants we speculate that the effect is likely to be substantially less than 23%. An analysis of licensure data suggests that the reduction in crashes may, in large part, be attributable to an overall reduction in exposure. PMID- 8703272 TI - Bicycle accidents and drivers' visual search at left and right turns. AB - The accident data base of the City of Helsinki shows that when drivers cross a cycle path as they enter a non-signalized intersection, the clearly dominant type of car-cycle crashes is that in which a cyclist comes from the right and the driver is turning right, in marked contrast to the cases with drivers turning left (Pasanen 1992; City of Helsinki, Traffic Planning Department, Report L4). This study first tested an explanation that drivers turning right simply focus their attention on the cars coming from the left-those coming from the right posing no threat to them-and fail to see the cyclist from the right early enough. Drivers' scanning behavior was studied at two T-intersections. Two well-hidden video cameras were used, one to measure the head movements of the approaching drivers and the other one to measure speed and distance from the cycle crossroad. The results supported the hypothesis: the drivers turning right scanned the right leg of the T-intersection less frequently and later than those turning left. Thus, it appears that drivers develop a visual scanning strategy which concentrates on detection of more frequent and major dangers but ignores and may even mask visual information on less frequent dangers. The second part of the study evaluated different countermeasures, including speed humps, in terms of drivers' visual search behavior. The results suggested that speed-reducing countermeasures changed drivers' visual search patterns in favor of the cyclists coming from the right, presumably at least in part due to the fact that drivers were simply provided with more time to focus on each direction. PMID- 8703273 TI - Enhanced tiredness among young impaired male nighttime drivers. AB - This study focuses on the incidence and amount of tiredness among young impaired male nighttime drivers during the hours between midnight and 6 a.m. One hundred and twenty impaired drivers, aged 18-30 years, with an average blood alcohol concentration of 1.54 g/l (range 0.81-2.99 g/l) are compared to 240 sober drivers of similar age stopped by the police in roadside surveys. A questionnaire shows that the increase in median awake period during the six observation hours is gradual and directly proportional to the time of the night for both groups, while the average awake period is significantly longer and that the average previous sleep period is significantly shorter for the impaired drivers. Self-assessment of tiredness shows 30 "rested," 68 "tired," and 22 "very tired" impaired drivers and 123 "rested," 105 "tired," and 12 "very tired" sober control drivers. This difference in level of "tiredness" is also significant. The questionnaire and the results of a simple visual reaction time test show individual differences for the drivers in both groups, but the difference in mean reaction time between the impaired drivers and the sober drivers is significant. Sub-division of the impaired drivers according to their self-assessed level of "tiredness" also shows significant differences in mean reaction time. The findings of the present study support the hypothesis that tiredness is an inherent and common accident risk factor among young male impaired drivers in nighttime traffic. PMID- 8703274 TI - An evaluation of road crash injury severity measures. AB - Reliable and consistent measures of injury severity are necessary for the study of environmental, crash and personal factors involved in road traffic crashes. This study was designed to evaluate measures of injury severity derived from computerized hospital discharge records, using 3609 road crash casualties admitted to hospital in Western Australia in 1988. External cause of injury codes were used to identify injuries from road traffic crashes. The ICDMAP software was used to convert the diagnosis codes into Abbreviated Injury Scale (AIS) scores for each body region. The maximum AIS values were derived using (1) all 12 diagnosis codes; (2) the first six diagnosis codes; and (3) the principal diagnosis code alone. Other measures of injury severity evaluated were the number of body regions with at least one injury; the number of regions with Abbreviated Injury Severity score of three or more; and total number of days spent in hospital. Discriminant analysis suggested that the AIS could be separated into minor and major injuries at a score of three and the Injury Severity Score at a score of nine. The measures derived from the AIS were all strongly correlated with each other and with the length of hospital stay and the dichotomized values gave similar results to the other scores when used in regression analyses of the injury experience of different types of road users. It was concluded that measures incorporating elements of both severity and number of injuries were preferable but length of hospital stay would be a suitable proxy if no other injury information was available. PMID- 8703275 TI - Differences in young driver crash involvement in states with varying licensure practices. AB - Teenage driver licensing practices and the crashes of teenagers were compared in several states with differing laws and policies regarding licensure. High school seniors in Delaware, a state with laws that allow early driving and licensure, reported that they first drove on a public road, obtained a learner's permit and obtained a driver's license at younger ages than high school seniors in other northeastern states (Connecticut, New Jersey and upstate New York). State crash data indicated that Delaware and Connecticut, which allow unrestricted licensing at age 16, showed the highest rates of 16-year-old driver involvements in nonfatal and fatal injury crashes. Pennsylvania and upstate New York, which have night driving curfews for 16 year-olds, showed lower crash rates overall and much lower crash rates during their respective curfew hours. New Jersey and Nassau and Suffolk counties, where unsupervised driving by 16 year-olds is generally not allowed, showed the lowest crash rates for 16 year-olds. Graduated licensing programs that include delayed full-privilege licensure, night driving curfews, and extended periods of supervised practice driving are a possible countermeasure for the high motor vehicle crash rates of young drivers. PMID- 8703276 TI - Estimating the consequences of accidents: the case of automatic train protection in Britain. AB - This paper considers estimates of the number of fatalities per year on rail lines operated by British Railways that could be expected to be saved by the installation of automatic train protection, based on historical data. The authors' preferred estimate is 3.66 fatalities per year, with an estimated 95% confidence interval of 1.44-5.89. It is possible to find orthodox probability distributions that fit the distribution of numbers of fatalities well. FN-graphs are not a suitable means of fitting such distributions. PMID- 8703277 TI - Helmets, injuries and cultural definitions: motorcycle injury in urban Indonesia. AB - This paper examines motorcycle helmet use and injuries in a developing country with a helmet law. Data were collected by systematic street observations and interviews with motorcyclists and supplemented with motorcycle injury data from a 1 month study of all patients coming to emergency departments in Yogyakarta, Indonesia. Observations show that 89% of motorcycle drivers (N = 9242) wore helmets; only 20% of the passengers (N = 3541) did. However, only 55% of the drivers wore helmets correctly (e.g. with chin strap buckled). Differences in time and place were noted in interviews when motorcyclists reported wearing helmets least at night and when no police were around; various reasons for not wearing helmets included physical discomfort and absence of police surveillance. Data from emergency departments found that motorcycles were involved in 64% of all traffic accident injuries, comprising 33% of total trauma patients presenting to emergency departments. Injury Severity Scores were calculated for the 26% of motorcycle injuries which were admitted to the hospital, with 60% having scores of 1-8, 27% 9-15, and 9% > 15. We conclude that although motorcycle drivers appear to comply with the motorcycle helmet law, it is a "token compliance." Less than 50% of riders were maximally protected by helmets and very little safety consciousness was found among drivers. Suggestions for improving helmet use that take cultural definitions of wearing helmets into account are presented for future research. PMID- 8703278 TI - Effectiveness estimates for center high mounted stop lamps: a six-year study. AB - The center high mounted stop lamp is a supplemental brake light mounted at the height of the rear window that has been required on all new cars sold in the U.S. starting with the 1986 model year. It was intended to reduce the number of rear end collisions by improving the braking signal recognition of following drivers. Insurance claims for vehicle damage were used to compare the rear-end collision rates of model year 1985 and model year 1986 cars over calendar years 1986-91. After adjusting for the difference in vehicle ages, model year 1986 cars were found to have experienced 5% fewer rear-end collisions than would have been expected without the center high mounted stop lamps. PMID- 8703279 TI - One-year economic consequences of accidents in a Swedish municipality. AB - The objectives of this study were to develop a system to calculate the economic consequences of accidents and to account for the economic consequences of all accidents during 1 year in a district. A total population injury survey was done in an area with a population of over 41,000. All accidents (N = 4926) occurring within a 12-month period and requiring medical care were noted. The costs (calculated at 1991 prices) to the health care service (outpatient care, including primary health care and hospital care) were SEK 23.7 million (US $3.59 million), to trade and industry SEK 79.7 million (US $12.08 million), and for health insurance SEK 9.1 million (US $1.38 million). The cost for society of uninsured people is not possible to estimate using loss of production. However, the time lost from "normal activities" was registered. If this time is valued at the same price as working hours, a welfare cost of SEK 43.1 million (US $6.53 million) should be added. In a forthcoming paper, an assessment of the cost effectiveness of interventions against accidents will be published. PMID- 8703280 TI - The influence of seat-back and head-restraint properties on the head-neck motion during rear-impact. AB - The influence of different seat properties on the head-neck motion during a low velocity rear-end impact was tested using a Hybrid III-dummy fitted with a modified neck (RID-neck). The results show that by modifying the properties of the seat-back and head-restraint it is possible to influence the head-neck kinematics to a great extent. It was possible to virtually eliminate the neck extension motion during a rear-impact. This will hopefully result in a significant decrease in neck injury risk in real world rear-impacts. PMID- 8703281 TI - Young novice drivers: towards a model of their accident involvement. AB - There are two main purposes of this article. The first is to present a model of young drivers' accident involvement, including the most important processes in the development of their driving behaviour. The model describes the relations between the learning process, the individual preconditions and the social influence and how these are related to driving behaviour and accident involvement through different processes such as skill acquisition, self assessment, information processing, feedback and motivation. The second purpose is to present a state-of-the-art report for the different aspects of the model by referring to some of the literature on young drivers. PMID- 8703282 TI - Young drivers' overestimation of their own skill--an experiment on the relation between training strategy and skill. AB - Young drivers' accident involvement may be explained by a number of different factors, one of which is that they tend to overestimate their skill in driving a car. This study is based upon the assumption that the degree of overestimation is related to the type of training the driver has received. In an experiment, two different strategies for training have been compared with regard to their influence on estimated and actual driving skill, as well as the drivers' degree of overestimation of their own skill. One of the strategies, used in the "skill" group was to make the learner as skilled as possible in handling a braking and avoidance manoeuvre in a critical situation. The other strategy, used in the "insight" group was to make the driver aware of the fact that his own skill in braking and avoidance in critical situations may be limited and unpredictable. The experiment was carried out at the Bromma driving practice area in Stockholm. Low friction has been simulated by using "Skid Car" equipment. Fifty-three learner drivers were randomly divided into two groups. Each of the groups was taught on the basis of one of the strategies. The training session was 30 minutes long. One week later, the drivers returned to take part in a test of their estimated and actual skill. The "skill" group estimated their skill higher than the "insight" group. No difference was found between the groups regarding their actual skill. The results confirm the main hypothesis that the skill training strategy produces more false overestimation than the insight training strategy. PMID- 8703283 TI - Spatial data aggregation: exploratory analysis of road accidents. AB - This paper is limited to the "size problem": it empirically and systematically examines to what extent the length of road segment influences the statistical description of accident counts and density. Univariate descriptive statistics are systematically measured for accident count and ratio, using different motorway segment lengths. A classification of the studied aggregation levels is proposed leading to three distinct groups of segments: (1) very small segments (1 hm), where observed accident counts are almost Poisson distributed, (2) large segments (more than 20 hm), where accident counts are almost normally distributed, and (3) medium size segments (3-20 hm), where accident counts have an intermediate empirical distribution. It is suggested that generalizations made at one level of spatial aggregation may not necessarily hold at another level. Conclusions derived at one scale may be invalid at another. The paper shows how important preliminary examination of the data can be, without any sophisticated statistical test. It also shows that one best or unique level of aggregation is not available: it depends upon the objective of the study. Results of any statistical analysis or modelling procedure should be seen as conditional on the scale of the analysis. PMID- 8703284 TI - Cellular phones and traffic accidents: an epidemiological approach. AB - Using epidemiological case-control design and logistic regression techniques, this study examined the association of cellular phone use in motor vehicles and traffic accident risk. The amount of time per month spent talking on a cellular phone and 18 other driver inattention factors were examined. Data were obtained from: (1) a case group of 100 randomly selected drivers involved in accidents within the past 2 years, and (2) a control group of 100 randomly selected licensed drivers not involved in accidents within the past 10 years. Groups were matched on geographic residence. Approximately 13% (N = 7) of the accident and 9% (N = 7) of the non-accident group reported use of cellular phones while driving. Data was obtained from Department of Motor Vehicles accident reports and survey information from study subjects. We hypothesized that increased use of cellular phones while driving was associated with increased odds of a traffic accident. Results indicated that talking more than 50 minutes per month on cellular phones in a vehicle was associated with a 5.59-fold increased risk in a traffic accident. The combined use of cellular phones and motor and cognitive activities while driving were also associated with increased traffic accident risk. Readers should be cautioned that this study: (1) consists of a small sample, (2) reveals statistical associations and not causal relationships, and (3) does not conclude that talking on cellular phones while driving is inherently dangerous. PMID- 8703285 TI - The feasibility of linking hospital and police road crash casualty records without names. AB - Government and other agencies routinely collect complementary information on road crash casualties and there may be opportunities to widen the scope of data available for research and policy evaluation if mechanisms can be established to combine these data sources. Where unit record data are available, direct links within and between individual records can often be achieved using record linkage techniques. Without the benefit of unique identifiers, sufficient other identifying information such as the full name, date of birth and sex of the casualty, their role in the crash and the date of the crash are required to enable correct links between individual records to be made. However, all these data items may not be available and it is therefore important to investigate the effect this may have on the quality and quantity of links between records. The research reported here used the hospital admission and police records from the Western Australian Road Injury Database, which had previously been linked using all the identifying characteristics mentioned above, special purpose software and comprehensive manual checking. This set of linked records were considered to be a "gold standard" which could be used to measure the quantity and quality of links produced using fewer identifying characteristics. Results from this process showed that about 90% of the original links could be identified when a phonetic code of the family name of the casualty was used with age, sex, road user type and crash date. However, only about 50% of the original links were found if linking was performed without using names or phonetic name codes. PMID- 8703286 TI - Cyclist road deaths in London 1985-1992: drivers, vehicles, manoeuvres and injuries. AB - We used coroners' records to investigate the fatal accidents of cyclists recorded on death certificates in London during 1985-1992. There were 124 deaths: 68 cyclists were injured in London and 56 injured in the "home counties" around London and died in London hospitals. The cyclists' ages were from 8-88, and 70% were male; the drivers were aged 17-74, and 96% were male. Of the 108 vehicles involved, 53 were cars (including one parked) and 40 heavy goods vehicles (HGVs) (including 14 large lorries and 18 articulated lorries), 5 light goods vehicles, 5 buses and coaches and 5 motorcycles. Fatal accidents occurred with a wide range of manoeuvres; of vehicles turning left (driving is on the left in the U.K.), 14 out of 15 were HGVs. Accidents were most often on 2 lane roads and one half were near a road junction. Law violations were recorded in half the accidents; alcohol intoxication contributed only rarely. While injuries to the head were the commonest reported direct cause of death, Inner London deaths were frequently due to multiple injuries. The study confirms the serious danger to cyclists (particularly women) in Inner London from large and articulated lorries, causing death from multiple injuries. PMID- 8703287 TI - Improving outcomes measurement through computer assisted documentation. PMID- 8703288 TI - Principles and techniques in the use of convexity. AB - Expected outcomes for a correctly fitted ostomy prosthetic device include sustained, predictable wearing time, protection of the stoma from injury, and maintenance of peristomal skin integrity. To accomplish this, the pouching system must be properly sized and maintain continuous contact with the peristomal skin. Continuous contact is achieved when the faceplate provides a mirror image of the topography of the peristomal plane. Varying degrees of support and convexity of the faceplate are used to fit the unique characteristics of the patient's peristomal plane and stoma. Support can be absent, soft or firm. Convexity is some degree of outward curving of the faceplate. Characteristics of the stoma (i.e., flush, retracted, loop), peristomal plane (i.e., creases, hernia, scarring) and abdomen (i.e., flaccid, soft, or firm) may indicate the need for convexity. Potential problems include mucocutaneous separation, ischemia, laceration, erythema, ecchymosis, and pain if convexity is used inappropriately. Alternatives to convexity include the use of an ostomy belt or surgery. Ongoing, intermittent assessment of the prosthetic is essential to evaluate that the convexity and support have achieved the expected outcomes. PMID- 8703289 TI - Contact Thermography in a study of primary healing of surgical wounds. AB - Occurrences in the wound until complete healing are manifested in different ways, one of which is temperature change. The aim of this study was establish if there is regularity in temperature changes in wounds healing by primary intention. Thirty examinees (27 female, 3 male) admitted for gallstone surgery were included. Temperature was measured for eight post-operative days at a fixed time each day and in the same room using the semi-quantitative, contact method of liquid crystal strips. The findings demonstrated that there were few differences among the collective thermal images of individual examines for any given post operative day. During the first 3 post-operative days, the temperatures rose, with few differences between the temperatures of the wounds and their wider surroundings. From day 4 through 8, the temperatures of the wounds and surroundings fell gradually. Stitches were removed on day 7, and on day 8 only the narrow zones of the incision sites were warmer than the surroundings. The findings suggest that there is regularity in the course of temperature of the surgical wound in primary healing. The clinical implications are that the persistence of a wider zone of increased temperature after day 4 predicts the possibility of wound infection and disturbed healing. PMID- 8703290 TI - Physical therapy modalities for wound management. AB - As part of a multidisciplinary team approach to the management of chronic wounds, physical therapists can add certain physical modalities to the care plan. Whirlpool, electrical stimulation, ultrasound, low-energy laser and compression therapy are physical therapy modalities that have been used to enhance wound healing. All of these modalities are used as adjunct treatments that, when appropriate, may help shorten the length of treatment and reduce patient suffering. Because the efficacy of some of these modalities remains to be established in controlled clinical trials, conventional wound care continues to be an important part of the team approach. PMID- 8703291 TI - Cost effectiveness in wound care. AB - Universal, objective definitions or formulas have not been available for calculating the costs of wounds and measuring cost/benefits of different care methods. Therefore, it is difficult to compare cost effectiveness of different treatments. A literature review revealed confusion concerning the differences between "cost" and "cost effectiveness". The costs of wound care are divided into direct costs (i.e., costs of supplies, surgical interventions, medications, inpatient care) and indirect costs (i.e., assistance with activities of daily living, days lost from work, litigation). Studies of cost effectiveness should measure cost per unit outcome. In wound care studies, outcomes are usually measured according to the goals of treatment. Common problems with published cost effectiveness studies are that cost is often confused with cost effectiveness; there is no standard method of calculating wound care costs; few studies have measured costs to achieve measured treatment outcomes; and outcomes are often measured or reported differently from study to study. In view of this, it is impossible to clarify cost effectiveness of healing, debridement, pain relief, etc. Until universal, objective scales to measure cost effectiveness are available, the clinician must read published studies critically to determine if treatment measures are indeed cost effective. PMID- 8703292 TI - Is a urinalysis necessary in assessing incontinence? PMID- 8703293 TI - Implementation of a comprehensive skin care program across care settings using the AHCPR pressure ulcer prevention and treatment guidelines. AB - Healthcare professionals in the Central Midwest identified the need for a comprehensive skin care program for pressure ulcer prevention and treatment across care settings. A multidisciplinary team, representing acute, extended and home care, was formed to create a program for all three settings based upon the AHCPR pressure ulcer guidelines. The team performed literature reviews on which to base the development and use of tools, conducted prevalence studies, and developed educational approaches. Implementation of the program was tailored for each setting. Some of the approaches used were a skin care fair, quality studies, continuous quality improvement concepts, a "Product Book" and educational presentations. Outcomes include improvement of continuity of care across settings and the use of the Braden Scale and the NPUAP pressure ulcer staging system. The focus has turned toward patient outcomes. Professionals have a better understanding of the care that is provided by other disciplines. Referrals are made based upon decision trees. Appropriate resources are used. Other outcomes anticipated include a decrease in nosocomial pressure ulcers, shortened wound healing time, appropriate referral of unresponsive chronic wounds, decreased discrepancies in wound documentation, decreased length of stay, improved financial outcomes, and improved client knowledge and participation. PMID- 8703294 TI - Approaches to venous leg ulcer care within the community: compression, pinch skin grafts and simple venous surgery. AB - The traditional approach to leg ulcer care is both expensive and achieves poor outcomes. The outcome of treatment for patients with venous ulceration has been improved using a more scientific and research-based approach to patient assessment, adequate compression bandaging, use of other procedures such as pinch skin grafting and simple venous surgery, and the delivery of these innovations in patient care to the community as a whole. Multi-layer compression bandaging is superior to standard (one-layer) bandaging because it can easily be adapted to a wide range of ankle circumferences and leg sizes to provide sustained, graduated compression. Pinch skin grafting is cost-effective, accelerates healing, and, following adequate training, may be performed by specialist nurses in the community. Simple venous surgery may be offered under local anaesthesia when only the superficial veins are incompetent; however, deep venous surgery has yet to prove itself in clinical practice. Implementation of dedicated community leg ulcer clinics using these research-based findings will result in a substantially reduced prevalence of venous leg ulceration. PMID- 8703295 TI - Co-factors in impaired wound healing. AB - Caring for individuals with chronic wounds is a complex process that requires a multi-disciplinary approach. Age, insufficient oxygenation/perfusion, malnutrition, bioburden, excess pressure, psychophysiological stress, concomitant conditions and adverse effects of treatment are the co-factors addressed in this manuscript. The evaluation of these co-factors is typically integrated into the initial history, physical, and ongoing assessment. Early identification of these co-factors allows the practitioner to establish a differential diagnosis, initiate proper referrals, and develop a comprehensive plan of care. Management of local and systemic co-factors that impact repair will mitigate their adverse effects and facilitate healing of the individual's chronic wound. PMID- 8703296 TI - Kudos to authors at the Community Leg Ulcer Clinic in London. PMID- 8703297 TI - ICWM statement on cost effective wound care. Evaluating your supply use to prepare for managed care. PMID- 8703298 TI - Preparing for managed care: taking the cost out of the supply system. PMID- 8703299 TI - NMDA receptor function and human cognition: the effects of ketamine in healthy volunteers. AB - A rapidly growing body of preclinical data has implicated the glutamatergic N methyl-d-aspartate (NMDA) receptor in memory and other cognitive processes. There is comparatively less information about this receptor system in human cognition. We examined the effects of subanesthetic doses of ketamine, a noncompetitive NMDA receptor antagonist, on two forms of memory, free recall and recognition, as well as attention and behavior in a double-blind, placebo-controlled, 1-hour infusion in 15 healthy volunteers. Ketamine produced decrements in free recall, recognition memory, and attention. In addition, ketamine induced a brief psychosis in our healthy volunteers marked by thought disorder and withdrawal retardation. Ketamine-induced memory impairments were not accounted for by changes in subject's attention and were not significantly related to psychosis ratings. These data suggest that the NMDA receptor plays a direct role in two types of explicit memory. The implications of these data for the pathophysiology of schizophrenia are discussed. PMID- 8703300 TI - Comparison of [3H]paroxetine and [3H]cyanoimipramine for quantitative measurement of serotonin transporter sites in human brain. AB - Previous studies have demonstrated that [3H]paroxetine and [3H]cyanoimipramine ([3H]CN-IMI) are highly selective ligands for the serotonin (5-HT) transporter. Using membrane preparation from the putamen, we confirmed that in human brain [3H]paroxetine labeled with high affinity one class of site associated with the 5 HT transporter. [3H]CN-IMI labeled two classes of sites in human brain. The one displaceable by 5-HT and with high affinity to 5-HT uptake inhibitors accounts for about 60% of the [3H]CN-IMI binding and, presumably, is associated with the 5 HT transporter. From the competition experiments, citalopram was selected to define [3H]CN-IMI binding to the 5-HT transporter in tissue sections because of its high selectivity to 5-HT transporter sites. A good correlation of the regional distribution patterns for [3H]CN-IMI and [3H]paroxetine was found using quantitative autoradiography. However, [3H]paroxetine underestimated high concentrations of the 5-HT transporter comparing to [3H]CN-IMI. This is likely to be due to the higher specific activity of [3H]CN-IMI. There is a good correlation between the regional distribution of 5-HT transporter sites labeled with either [3H]paroxetine or [3H]CN-IMI and the density of serotonergic innervation. This suggests that the brain areas that receive numerous serotonergic afferents, such as the hypothalamus and basal forebrain, might be common targets of these antidepressant drugs. Pharmacologic similarity of the sites labeled by both ligands as well as their similar distribution in the brain suggests that both antidepressant drugs interact with the same protein, thereby eliciting a similar neurochemical response. PMID- 8703302 TI - Intact 5-HT2A receptor exons and the adjoining intron regions in schizophrenia. AB - Genes that regulate serotonergic (5-HT) systems may underlie the etiology of schizophrenia. In this study the gene encoding the 5-HT2A receptor in schizophrenics and healthy controls was examined. First, we sequenced all exons and the flanking introns of the 5-HT2A receptor gene in 10 schizophrenics and 10 controls. The substitution of C for T at position 102 in exon, which had been reported by Warren et al. (1993), was confirmed. Restriction fragment length polymorphism (RFLP) analysis revealed no association between polymorphism and schizophrenia. There was no association between the polymorphism and subdiagnosis, family history, age of onset, amounts of antipsychotics, or positive and negative symptoms before or after medication. Other polymorphisms in the gene were screened in 100 schizophrenics by the single-strand conformation polymorphism method, but none was found. Our results suggest that an abnormality in the 5-HT2A receptor gene in schizophrenia is unlikely. PMID- 8703303 TI - Reduced prolactin and cortisol responses to d-fenfluramine in depressed compared to healthy matched control subjects. AB - d-Fenfluramine, a specific 5-HT releasing agent without the catecholamine effects of d,l-fenfluramine, was used as a neuroendocrine challenge in 19 subjects with major depression and 19 healthy controls. Patients and controls were matched for age, sex, weight, and menstrual status. 5-HT-mediated prolactin and cortisol responses were both significantly attenuated in the depressed group. Patients with a history of a suicide attempt had lower cortisol responses than those without. Peak cortisol responses were inversely related to baseline cortisol levels. There were also significant relationships between hormone responses and both age and weight. These findings replicate those of a previous study using this challenge and reiterate the role of reduced 5-HT activity in suicide. They also reinforce the need for careful matching in neuroendocrine studies. PMID- 8703301 TI - Regional differences in the effects of amphetamine withdrawal on dopamine dynamics in the striatum. Analysis of circadian patterns using automated on-line microdialysis. AB - The purpose of the study is to determine the relationship between behavioral symptoms of amphetamine withdrawal and the extracellular concentration of dopamine (DA) in the dorsolateral caudate nucleus and the nucleus accumbens across the entire light-dark cycle. This was accomplished using automated on-line microdialysis sampling in behaving rats. Animals were pretreated with escalating doses of d-amphetamine (or saline) over a 6-week period and then were withdrawn from amphetamine for 3, 7, or 28 days before testing. There were regional differences in the effects of amphetamine withdrawal on the concentrations of DA and DA metabolites in dialysate. Early during withdrawal (3 and 7 days), when animals showed postamphetamine withdrawal behavioral depression (nocturnal hypoactivity), there was a significant decrease in DA and DA metabolites in the dorsolateral caudate nucleus and a disruption in the normal circadian pattern of DA activity. In contrast, there was no effect of amphetamine withdrawal on DA dynamics in the nucleus accumbens. By 28 days after the discontinuation of amphetamine pretreatment, after basal DA in the caudate returned to normal, there was a significant increase in basal DA metabolism in both the caudate and the accumbens. This increase in DA metabolism may be related to the expression of sensitization, including a hypersensitivity to an amphetamine challenge. It is concluded that the role of the dorsal striatum in psychostimulant drug withdrawal syndromes deserves further consideration. PMID- 8703304 TI - Symptoms essential to the experience of sodium lactate-induced panic. AB - We report on subjective ratings and symptoms experienced before and during sodium lactate infusion by patients with panic disorder or agoraphobics with panic attack (DMS-III-R criteria). Symptoms were assessed using the Acute Panic Inventory (API). During the lactate infusion 59% of the patients were rated by an attending psychiatrist as having experienced lactate-induced panic attacks. Patients experiencing lactate-induced panic attacks overwhelmingly rated this experience as very similar to their typical naturally occurring attacks. Among the individual API symptoms items at baseline (prelactate) only Afraid in general (r = 0.26) was significantly, but not strongly, correlated with the panic response. Controlling for baseline symptom levels, the most robust partial correlations of symptomatic increment with panic were Desire to flee (0.70), Fear of losing control (0.57), Afraid in general (0.49), and Dyspnea (0.48). Using a dichotomized symptom increment greater than 1, 13 of 29 API items indicated a panic response to lactate infusion; the best were Dyspnea, Feeling confused, Afraid in general, Difficulty speaking, Difficulty concentrating, Desire to flee, and Fear of losing control. A logistic regression analysis showed that among baseline measures, Afraid in general and Feeling confused significantly predicted panic. For dichotomized change scores, Afraid in general, Dyspnea, and Dizziness/lightheadedness significantly indicated panic. In these analyses three symptom items stand out as the most predictive and revealing of panic to lactate infusion: Afraid in general, Dyspnea, and Desire to flee. These results are discussed in the context of Klein's (1993) suffocation false alarm theory of panic. PMID- 8703305 TI - Effect of dimaprit and cimetidine on the somatostatinergic system in the rat frontoparietal cortex. AB - A recent study carried out by this laboratory demonstrated that exogenous histamine increases the somatostatin (SS) receptor/effector system in the rat frontoparietal cortex (Puebla and Arilla, 1995). In the present study we examined the participation of the H2-histaminergic system in this modulation by use of the H2-receptor agonist and antagonist dimaprit and cimetidine, respectively. Dimaprit administration [20 micrograms/rat, intracerebroventricularly (ICV)] to rats 2 hours before decapitation increased the number of SS receptors in the frontoparietal cortex without changing the affinity constant. Pretreatment with cimetidine (20 micrograms/rat, ICV) prevented the dimaprit-induced changes in SS binding in the frontoparietal cortex, whereas cimetidine alone (20 micrograms/rat, ICV) had no observable effect on this parameter. The in vitro addition of dimaprit or cimetidine to frontoparietal cortex membranes from untreated rats did not markedly affect the SS binding characteristics. Somatostatin caused a significantly higher inhibition of basal and forskolin (FK) stimulated adenylyl cyclase (AC) activity in frontoparietal cortex membranes from dimaprit-treated rats than in controls, an effect that was prevented by pretreatment with cimetidine. No significant differences, however, were detected for the basal or FK-stimulated AC enzyme activity in the control, dimaprit-, and/or cimetidine-treated groups, which suggests no impairment of the AC catalytic subunit. In addition, the functional activity of the guanine nucleotide binding inhibitory protein Gi, as measured by the capacity of the stable GTP analogue 5'-guanylylimidodiphosphate [Gpp(NH)p] to inhibit FK-stimulated AC activity, was not altered by dimaprit. Thus, the increased SS-mediated inhibition of AC activity observed in the dimaprit-treated rats may be caused by the increase in the number of SS receptors. Neither dimaprit nor cimetidine affected somatostatinlike immunoreactivity (SSLI) content. The present results, together with the fact that SS and histamine have been shown to influence locomotor activity and nociception in a similar manner, suggest that some of the neurotransmitter effects of SS may be modulated by histamine via H2-histaminergic receptors. PMID- 8703306 TI - Adhesion molecules in allergy. AB - Cell adhesion molecules have been recognized to play a major role in a variety of physiological and pathophysiological phenomena, such as embryogenesis, maintenance of tissue architecture and tissue damage repair, recruitment of leukocytes into tissues, immunological reactions requiring cell-to-cell contact. Their role in inflammatory reactions is crucial, in particular their involvement in allergic inflammation has been extensively studied. It has become apparent that the expression patterns and cell localization of certain adhesion molecules are related to the dynamic of allergic inflammation, and specifically to selective migration of eosinophils and interactions with epithelial cells of the target organs. In some instances, cell adhesion molecules by themselves may be regarded as reliable markers for clinical purposes. In addition, the pharmacological modulation of expression and release of adhesion molecules may provide new insight about the mechanisms of action and effectiveness of antiallergic drugs. PMID- 8703307 TI - Effect of conditions in obtaining blood samples for ECP testing in children. AB - Eosinophil Cationic Protein (ECP) is a basic protein found in eosinophil granules. This cell and its mediators are currently considered to be potential indicators of the severity of inflammation in the organism. ECP concentration can be reliably tested using several RIA or ELISA methods. It is well known that the conditions of sample obtention can affect the ECP values in blood. The aim of this study is to establish which parameters affect ECP testing during regular blood sample collection and how they affect it. Blood samples taken for the routine study of five children attended in our department were analysed: four were asthmatic and one child had atopic dermatitis. In the results we observed that ECP was not detected in the blood samples taken with EDTA tripotassium. In both the plasma samples taken with heparin as well as with serum, more ECP was released at a higher temperature. In the release of ECP obtained by coagulation, samples at 37 degrees showed values of between 4 and 20 higher than those obtained for an hour at 0 degrees. There is a considerable variability in the testing of ECP depending on the blood test extraction conditions, the range is bigger in the samples with eosinophils. These results imply the need to define a stricter protocol for obtaining samples than that suggested at present. PMID- 8703309 TI - [Platelet antigens: immunology and immuno-allergology]. AB - Platelet immunology allows the understanding of clinical findings in a genetic and serologic basis. Blood platelets bear common antigens and same specific antigens, classified in five groups (HPA 1 to 5), that are localized on membrane glycoproteins Ia, Ib alpha, IIb and IIIa. Antiplatelet autoimmunization is generally due to IgG antibodies against membrane complexes IIb/IIIa or Ib/lX. Antiplatelet alloimmunization, clinically resulting in Posttransfusion Purpura and Neonatal Thrombocytopenia is more frequently associated with anti-IIb/IIIa antibodies, either anti-HPA-1a or HPA-1b. Finally, platelet participation in immunoallergic reactions is discussed, focusing both platelet activation by allergen itself and platelet recruitment by other inflammatory cells. PMID- 8703308 TI - [Importance of the leukotriene C4 liberation test for the diagnosis of drug allergy (preliminary results)]. AB - Pharmacological or immunological activation of human basophil induces the expression of membrane markers such as CD63 and the release of histamine and leukotriene C4 (LTC4). LTC4 release test has the advantages of a cellular test which is obviously closer to the physio-pathological reaction than the direct determination of circulating antibodies. Moreover, the determination of a newly formed mediator eliminates the spontaneous release of mediator by simple cellular damage. We present here, after having controlled the reliability of the method for aero-allergens, preliminary results of LTC4 release obtained for haptens (ampicillin, methylprednisolone and acetylsalicylic acid) which are in favour of the use of this method in the complex field of drug allergy diagnosis. PMID- 8703310 TI - [Do nasal and conjunctival provocation tests represent a trustworthy parameter for the follow-up of childhood IgE-dependent bronchial asthma?]. AB - The aim of this study is the evaluation of the response at the level of different organs (nose and conjunctiva) of desensitizing treatment in children suffering from bronchial asthma and receiving immunotherapy with D. pteronyssinus. PMID- 8703311 TI - Study on the ability of a new barrier to reduce airborne mite allergens. AB - Environmental control, especially means for blocking the release into the air of mite allergens embedded in padded furniture (mattress, pillows), represents the first important step in the treatment of IgE-mediated allergic diseases. New to the market is a home hygiene spray that creates a protective barrier on padded furniture and is thereby able to contain the dispersion into the air of allergic particles. We tested the effectiveness of the spray in vitro. Since mites, especially D. Pteronyssinus, are considered the foremost allergens, we verified the ability of the invisible barrier to impede the diffusion of the allergen and the mobile forms of these mites (Der p 1, Der 2). The product under exam created a barrier that totally blocked the passage of mobile forms and almost totally prevented the passage of allergens (> 99%). PMID- 8703312 TI - [Conjunctivitis, keratitis, keratoconjunctivitis: role of allergy]. AB - The author has counted 0.85% of conjunctivitis in an unselected allergic population of 2,865. All the causes are explored, microbial, viral, metabolic , ophthalmological, drugs, mechanical, allergic... He describes different forms of conjunctivitis amongst which are those of spring time. The roles of the ophthalmologist and allergist are emphasized for establishment of diagnosis and treatment. PMID- 8703313 TI - Orally administered bacterial lysate Broncho-Vaxom for the treatment of common variable immunodeficiency. AB - Broncho-Vaxom (B-V) is a lysate of eight bacterial pathogens of the respiratory tract with immunomodulatory properties. It is used in prophylaxis of respiratory tract infections. We conducted an open, placebo controlled, cross-over trial of B V treatment in patients with common variable immunodeficiency (CVID). B-V or placebo were given for the period of four months either in winter or spring period. No significant improvement of clinical state of patients during the period of B-V treatment compared with the period when placebo was administered was observed. When subjective comparison of the health state of patients with the same period of the previous year was assessed, significant improvement after B-V treatment compared to placebo was recorded. Evaluating serum immunoglobulin levels a significant increase in serum IgA level was observed after B-V treatment. The results warrant further studies of B-V treatment in CVID patients. PMID- 8703314 TI - [Overview on RAST inhibition]. AB - The technique of RAST inhibition is always useful in two cases: to study cross reactions between the allergens; for more specific use of allergen indicated for desensitization. The comparison made in this article concerns dog allergens. As can be seen in the tables, diagrams and graphs, RAST inhibition by the dog allergen from the laboratories of DHS-Bayer-Pharma was clearer than that from the Laboratoire Stallergenes. PMID- 8703315 TI - [Anaphylactic reaction to Aspergillus]. AB - Nowadays the pollens are the principal allergens implicated in anaphylactic reactions. Now one of our observations is of a case history of recurring acute urticaria, with malaise, hypotension, Quincke's oedema and dyspnea, linked with Aspergilus. The etiology is based on clinical observation, skin tests and specific IgE with a significant atopy. This observation shows that the anaphylactic reaction can also be associated with a mould such as Aspergillus. PMID- 8703316 TI - Dosimetry and treatment planning of complex electron arc therapy. AB - Electron arc therapy has been primarily utilized for treatment of the post mastectomy chest wall. Its clinical usefulness and benefits have been proven. Casting procedures and blocking techniques have been discussed. We have successfully applied electron arc therapy to the treatment of the nasal cavity and associated nodal regions. When compared to the chest wall, the smaller radii and pronounced irregularity of the surface anatomy of the facial region presents multiple new dosimetric and treatment planning challenges. We will discuss these challenges, along with those encountered in the casting procedures and reproduction of patient characteristics required for the execution of this treatment. Many aspects of dosimetry were utilized in unique ways to produce the treatment plans, cast and bolus needed. Due to the insertion of bolus into the nostrils, a unique hollow bite block was constructed to allow patient respiration during treatment. Film dosimetry was used to verify computer predictions and to compare this electron arc treatment technique with alternative fixed electron beam techniques. The potential benefits and difficulties of this technique will be discussed. PMID- 8703317 TI - An asymmetric half-wedged field technique for the treatment of inferiorly extended head and neck cancers. AB - Adequate dose coverage of the target volume of head and neck cancer patients can be difficult to achieve with lateral opposing fields when the patient's neck is short and the disease involves the lower cervical region. Rotation of the couch away from the side of the gantry, such that the beams are angled towards the inferior aspect of the patient, can result in a better dose distribution. However, if the neck separation is large, unacceptable dose inhomogeneities will still exist. The conventional approach in these situations is to treat with parallel opposed lateral fields junctioned with anterior/posterior opposed low neck fields. We present a variation on the couch rotation technique as an alternative. Asymmetric jaws and half-wedged fields are used to obtain a homogeneous dose within an inferiorly extended target volume. The advantages of this technique are a simple set up and the capability of adding spinal cord shielding without shielding midline primary tumour. PMID- 8703318 TI - A method for total skin electron treatment for infants. AB - Diseases such as mycosis fungoides require the treatment of a patient's total skin surface with superficial radiation. In a unique clinical situation, a 14 month-old child presented with a need for total skin treatment. A typical total skin technique requires overlapping electron beams, using 6 body positions, each with the gantry rotated for 2 angulations, or '6 positions-12 fields'. Adaptation of this technique for infants is complicated by the small diameter of some body parts, and by the necessity to treat while the patient is anesthetized. Even degraded, low energy electrons can easily penetrate fingers and toes. Therefore, dose from 6 positions becomes additive, and the total dose to small circumferences can be 3 to 4 times more than skin dose on the torso, raising concerns about uneven bone growth in the developing child. Special phantoms were designed for extensive dosimetry needed to determine both dose rate and dose summation from the overlapping beams. Computerized electron pencil beam calculations were compared to TLD measurements. Unique compensating techniques were used to deliver uniform dose. A modification of the 6 position-12 field technique will be described; and accessories used to reduce high dose regions will be illustrated. PMID- 8703319 TI - Abutment of adjacent intraoperative radiotherapy electron fields to treat large tumor volumes. AB - Abutment of unmodified Intraoperative Radiotherapy (IORT) electron fields to irradiate large volumes can lead to dose inhomogeneities at the junction site of the matched fields. Although precise field matching is difficult to achieve in the IORT setting, we have fabricated special cones and dosimetrically evaluated their use. Based on these results, we have established guidelines for the routine use of multiple IORT fields that require field matching. This study examines the abutment of adjacent IORT electron fields for various cone sizes and beam energy combinations, using film dosimetry and a scanning densitometer. Results indicate that for abutting two adjacent 9 x 9 cm square cones, an optimum field separation at the depth of the 90% isodose line (d90) can be obtained as follows: For 6, 9, and 12-MeV electron beams, abut the inner wall of the second IORT cone to the inner wall of the first IORT cone. For 16- and 20-MeV electron beams, abut the inner wall of the second IORT cone to outer wall of the first IORT cone. PMID- 8703320 TI - Software control procedures for treatment planning systems. AB - In radiotherapy treatment planning systems, the software programs as well as the beam data have to be updated frequently, when new software versions are released or as beam data change. This update procedure has to be carried out carefully, to ensure the integrity of the system software and data. Moreover, the version of software used for a particular patient treatment plan is important, not only as a record to be retrieved when necessary, but also when there are multiple terminals connected to the same computer. This process of version control is not always carried out in a systematic and standardized fashion. Most clinics often have their own systems to implement this procedure, but while some systems are comprehensive, others may not necessarily incorporate enough safeguards for errors. In addition, treatment planning system manufacturers often do not offer well-designed and fail-safe facilities for this important issue. This report describes the software update control procedures we have implemented on our treatment planning system as well as suggests some general principles that could be applied to other planning systems. PMID- 8703321 TI - Sensitivity and stability of LiF thermoluminescence dosimeters. AB - Three different batches of LiF TLD-100 rods were subjected to investigation of their radiation sensitivity and the stability of the sensitivity after subsequent cycles of irradiation readouts. An old batch (circular rods from 1985), R85, showed an initial sensitivity of approximately 720 (nC/Gy). A new batch of circular rods (R0059) had an initial sensitivity more than a factor of 2.5 higher than the old batch. A batch of square rods (S3720) had an even higher sensitivity; a factor of almost 4 higher than batch R85. Both batches of circular rods showed a statistically significant change in sensitivity with increasing cycles of irradiations and readouts. No significant change was observed for the batch S3720. Batch R85 showed an increasing sensitivity with increasing number of cycles, and a small reduction in the width of the sensitivity distribution within the batch. However, the newer batch (R0059), showed a marked decrease in sensitivity and a doubling of the broadening of the sensitivity distribution after 10 cycles. No significant change in the sensitivity distribution within the S3720 batch was observed after 10 cycles. PMID- 8703322 TI - Protection of testes during radiation treatment by irregular and focused fields of 25 MV x-rays: in vivo evaluation of the absorbed dose. AB - During therapeutic irradiation of subdiaphragmatic nodal areas, the gonads are exposed to radiation from the primary beam and scatter. Since young patients have a high probability of cure, limiting exposure to the gonads should be pursued. Primary shielding may be supplemented by additional shields in order to reduce this exposure. Testes dose measurements were performed with thermoluminescent dosimetry (TLD) in 37 patients treated with 25 MV x-rays on subdiaphragmatic nodal areas. Each patient was measured 5 times while under treatment. In 27 cases, an additional shield was placed to protect the testes. For 9 cases, comparative measurements were performed on a phantom with and without additional shielding. The median dose received by gonads was respectively 3% with additional shielding (27 cases) and 5.8% without additional shielding (10 cases, p = 0.001 Mann Whitney test). In the 6 patients for whom the measurements were compared, the differences were also statistically significant (p = 0.028 paired Wilcoxon test). This study confirmed the benefit of additional gonadal shielding during subdiaphragmatic radiation treatment with 25 MV x-rays. PMID- 8703323 TI - An equivalent square method for irregular photon fields. AB - A quick formula is proposed for calculating equivalent squares of irregularly shaped photon fields. No complex calculations are required. Three different energies (60Co; 6 MV and 18 MV x-rays) were investigated. Comparison with results utilizing the Clarkson technique (60Co) and with experimental measurements (6 MV and 18 MV x-rays) indicates that the method is accettable in a wide variety of clinical situations. The applied procedure introduces an indetermination lower than 4%, with maximum absolute percentage errors of 3%, 2.7%, and 3.6% for 6 MV, 18 MV, and 60Co photon fields respectively. PMID- 8703324 TI - Off-axis point localization for mantle fields: the St. Jude experience. AB - Mantle field irradiation is a common occurence in pediatric settings. At St. Jude Children's Research Hospital, precise locations have been determined for the five off-axis points which are routinely used for mantle field calculations at our institution. This has helped to develop consistency between mantle field patients regarding off-axis point localization. PMID- 8703325 TI - Evaluation of dose delivery based on a comparison of dosimetry calculations using open beam and wedged beam depth dose data. AB - The purpose of this study is to evaluate the magnitude of the error in dose delivery caused by the use of open beam depth dose data in dosimetry calculations for wedged photon beams. Isodose plans were calculated for treatments given in a 3-field isocentric prostate or rectal setup using an open AP beam with two lateral wedged beams. The dose distributions were first calculated using open beam depth dose data for all three fields. Next, the open beam data was used only for the AP field and true wedged beam depth dose data was substituted for the two lateral wedged fields. The magnitude of the depth dose variations for wedged vs open beams depends on the nominal beam energy, the wedge angle, and the depth of measurement. Consequently, isodose distributions calculated for wedged fields were found to be different when true wedged beam depth dose data was used instead of open beam data as is commonly done. Monitor unit calculations using a field size specific wedge factor show that dose delivery errors up to 4% can result from the use of open beam depth dose data in wedged beam dose distribution calculations for a 6-MV photon beam. Accurate treatment planning for wedged fields requires the use of wedged beam depth dose data specific to each wedge. Simply using open beam depth dose data in dose calculations for wedged beams will result in dose delivery errors, the magnitude of which depends on the combination of wedge angle, field size, and nominal beam energy. PMID- 8703326 TI - Evaluating the dose to the contralateral breast when using a dynamic wedge versus a regular wedge. AB - The incidence of secondary cancers in the contralateral breast after primary breast irradiation is several times higher than the incidence of first time breast cancer. Studies have shown that the scatter radiation to the contralateral breast may play a large part in the induction of secondary breast cancers. Factors that may contribute to the contralateral breast dose may include the use of blocks, the orientation of the field, and wedges. Reports have shown that the use of regular wedges, particularly for the medial tangential field, gives a significantly higher dose to the contralateral breast compared to an open field. This paper compares the peripheral dose outside the field using a regular wedge, a dynamic wedge, and an open field technique. The data collected consisted of measurements taken with patients, solid water and a Rando phantom using a Varian 2300CD linear accelerator. Ion chambers, thermoluminescent dosimeters (TLD), diodes, and films were the primary means for collecting the data. The measurements show that the peripheral dose outside the field using a dynamic wedge is close to that of open fields, and significantly lower than that of regular wedges. This information indicates that when using a medial wedge, a dynamic wedge should be used. PMID- 8703328 TI - Dose to contralateral breast--a comparative study. AB - Radiation, especially subtherapeutic doses, has been proven to be carcinogenic. During therapeutic irradiation, normal tissue is exposed to some dose due to internal and external scatter from the primary beam. The dose to the contralateral breast during primary irradiation may cause secondary breast malignancies. For example, studies have shown that half-beam block techniques can dramatically increase the opposite breast dose. An attempt has been made to calculate the dose delivered during primary breast irradiation. Materials used will include previously published articles in medical and science journals. Preliminary results from a study currently being done by Marilyn Stovall will also be utilized. Isodose distributions demonstrating various treatment techniques will be shown. PMID- 8703327 TI - Influence of inguinal node anatomy on radiation therapy techniques. AB - A 1992 GOG (Gynecologic Oncology Group) study suggested that groin irradiation does not control microscopic inguinal node metastases as well as does surgical dissection. That trial has come under some scrutiny, as possible deficiencies in the radiation regimen used might have influenced results. The study received criticism for assigning a prescription point [Rx@3 cm below anterior skin surface] to patients which may not have given adequate coverage to the inguinal nodes. The inguinal node depths of 31 patients with cervical, vaginal, or vulvar malignancies were measured from their planning CT scans and then compared to depth of the prescription point designated by the radiation therapists in the aforementioned study. Twenty-four of eighty-one viable superficial inguinal node depth measurements were greater than 3 cm, and all of eighty-four deep inguinal node measurements were outside the three centimeter range. The results of this project would indicate that the depths of patients' inguinal nodes vary enough to justify alterations in some present external beam radiation therapy techniques. Indiscriminate prescription points will no longer suffice if groin radiation is to be effective, and treatment planning must utilize imaging studies to devise radiation regimens that provide optimal dose to the superficial and deep inguinal nodes. PMID- 8703329 TI - [Progress report for the period 1 January 1994 - 31 December 1994]. PMID- 8703330 TI - [Advances in the study on artificial cultivation of Cordyceps sinensis]. PMID- 8703331 TI - [Textual study on traditional Chinese drugs: haima and hailong]. PMID- 8703332 TI - [Preliminary survey on the traditional Chinese drug: shell of abalone or sea ear]. AB - According to an on-site survey of the commercial medicinal materials in China, it is demonstrated that the shells of abalone or sea-ear consist of nine types. Their original animals are Haliotios diversicolor, H. discus hannai, H. ovina, H. asinia, H. ruber, H. laevigata, H. semistrata, H. midae and H. cracherodii. Three of these types namely, H. semistrata, H. midae and H. cracherodii are not included in the Chinese Pharmacopeia. Furthermore, most of the shells of abalone or sea-ear presently on market are Haliotios ruber and H. discus hannai. PMID- 8703333 TI - [Preliminary study on physiology and biochemistry in Fritillaria thunbergii Miq. in the alternate process of new and old organs]. AB - The changes of weights, various sugars and diastase of the old scale and bud in Fritillaria thunbergii were studied in the alternate process of the new and old organs. The results showed that starch was finally disintegrated and inverted to sucrose before being transported to the new organ. PMID- 8703334 TI - [Technology of processing carbonized root of Sanguisorba of ficinalis L. and the quality standards for its prepared pieces]. AB - The technology of processing carbonized root of Sanguisorba of ficinalis was selected using orthogonal experiment design. The result shows that the best way is to stir-fry the drug in a pan for 7.5 min at 250 degrees C. The contents of trace elements in the prepared piece are significantly increased, and the prepared piece have marked hemostatic and bacteriostastic effects. PMID- 8703335 TI - [Preparation of oral berberine bisulfate liposomes]. AB - The co-precipitate has been made from berberine bisulfate and polyvingyl pyrrolidone (PVP) with the ratio of 1 : 5. The liposomes entrapped berberine bisulfate have been prepared with the ration of drug to lipid 1 : 20 and the ratio of cholesterol to soy phosphatide 2 : 7. The percentage of encapsulation is 48.72%. The test in vivo of small intestinal absorption in rats shows that liposomes can increase the absorption by 4-fold odd compared with free ber-berine bisulfate. PMID- 8703336 TI - [Determination of panaxadiol and panaxatriol in shihu yeguang pills]. AB - The contents of panaxadiol and panaxatriol in Shihu Yeguang Pills manufactured by pharmaceutical works in Beijing, Jinan, Guangzhou and other places were determined by TLC-densitometry. A quantitative method has been developed to serve as a reference to quality control of these pills. PMID- 8703337 TI - [Chemical constituents of Stellera chamaejasme L]. AB - Twelve compounds were isolated from the roots of Stellera chamaejasma. Five of them were elucidated by spectroscopic and chemical methods as 3', 14-dimethyl-4', 11-dimethoxy-5, 7-dihydroxybenzoflavanone (10), daphnetin (1), umbelliferone (2), daucosterol (5) and beta-sitosterol (3). Compound 10 is new. Compounds 1, 2, 5, 10 were isolated for the first time from S. chamaejasme. PMID- 8703338 TI - [Chemical constituents of Pleurospermum govanianum (Wall) Benth ex C. B. Clarke var. bicolor Wolff]. PMID- 8703339 TI - [Chemical composition of Pleurospermym rivulorum (Diels)]. PMID- 8703340 TI - [Chemical constituents of Euphorbia hylonoma Hand. -Mazz]. AB - Three compounds have been isolated from the root of Euphorbia hylonoma and identified as 3,3',4-tri-O- methylellagic acid, beta-sitosterol and sucrose by physicochemical and spectroscopic methods. The compound 3,3'4-tri-O-methylellagic acid is discovered in Euphorbia for the first time. PMID- 8703341 TI - [Protective effects of liu tea on CCl4 injured liver and its working mechanism]. AB - Liu Tea can effectively prevent the elevation of SGPT and SGOT induced by CCl4 in mice, reduce MDA and elevate GSH and GSH-px levels. It also induces the anabolism of microsomal protein and hepatocyte microsomal cytochrome P-450. These results suggest that Liu Tea has protective effects on the liver, which may be related to the clearing of free radicals and inhibition of lipid peroxidation. PMID- 8703342 TI - [Scavenging effects of daphnetin and its Cu, Zn complexes on superoxide radical]. AB - The scavenging effects of daphnetin (D) and its Cu, Zn complexes on superoxide radical (O2-.) generated through the photooxidation of riboflavin were studied with human red blood cells (RBC) and RBC membrane as experimental material. The Cu ( II ) complex showed the highest activity. SOD, D and its Cu ( II ), Zn ( II ) complexes were found to have inhibitory effect on the production of lipid peroxide in the membrane, SOD being the best among them. PMID- 8703343 TI - [Antifebrile and anti-inflammatory effects of radix Cynanchi atrati]. AB - The water extract of Radix Cynanchi Atrati used as intraperitoneal injection has been proved to have an obvious antifebrile++ effect on rat fever caused by 15% yeast suspension hypodermic injection as well as a significant anti-inflammatory effect. But the antiferbrile effect of its ethanol extract is not clear. PMID- 8703344 TI - [Type B supersonic diagnostic observation on the action of 25 kinds of traditional Chinese medicine on gallbladder movement]. AB - Twenty-five kinds of traditional Chinese medicine (TCM), which are often used in clinic to cure cholelithiasis, were studied for their actions on gallbladder movement in cholelithiasis patients by using a type B supersonic diagnostic set. The results show that among these 25 kinds 15 were of contracting type, 4 were of diastole type and 6 were of contracting and diastole type, thus providing useful experimental criteria for the clinical application of these TCM in cholelithiasis patients. PMID- 8703345 TI - [Restricting effects of geologic background system on genuine crude drugs in Sichuan]. AB - The distribution, growth, output and quality of genuine crude drugs are restricted by geologic background system (GBS), whose extensional vector system "rock-->soil-->medicinal plants" accomplishes the unity of geological grand cycle and biological pulmonary circulation. This article describes how genuine crude drugs in Sichuan, such as Coptis chinensis, etc. for example, are restricted by GBS. PMID- 8703346 TI - [Experiment on the rapid reproduction of Fritillaria taipaiensis P.Y. Li]. AB - This paper reports the experimental results of rapid reproduction of Fritillaria taipaiensis by tissue culture. The research shows that the induction frequency of calluses and rebulbs with MS+NAA lmg/L+6BA 3mg/L is 93%. Fifteen kinds of common and trace elements that are necessary for human body are found in calluses and rebulbs. The contents of most elements obviously exceed those of cultural bulbs. PMID- 8703347 TI - [Determination of paeonol in carbonized bark of Paeonia suffruticosa Andr. by HPLC]. AB - The contents of paeonol in prepared pieces of carbonized bark of Paeonia suf fruticosa were determined by HPLC. The result shows that the contents of paeonol decrease with the rise of temperature and the extension of time in processing. PMID- 8703348 TI - [Quality control of zhuanggu shenjin capsules]. AB - Studies have been made on the microscopic identification of red jinseng, dog bone, indian bread, yanhusuo and datura flower in Zhuanggu Shenjin Capsules, as well as on the identification of ginsengside and tetrahydropalmatine by TLC and the determination of icariin by dual wavelength TLC-scanning. The content of the latter compound should be no less than 0.380mg/g and the average rate of recovery 97.62% (n = 5). PMID- 8703349 TI - [Chemical constituents of Taxillus levinei (Merr.) H.S.Kiu]. AB - Four compounds were isolated from the leaves of Taxillus levinei and identified as protocatechuic acid, isoquercitrin, quercetin-3-O-(6"-galloyl)-beta-D glucoside and quercetin-3-O-beta-D-glucuronide on the basis of chemical properties and spectral data. These compounds were obtained from T. levinei for the first time. PMID- 8703350 TI - [Iridoid glucosides from Phlomis mongolica Turcz]. PMID- 8703351 TI - [Chemical components of Lindernia ciliata (Colsm) Pennell]. AB - Three known compounds were isolated from the petroleum ether extract of the whole plant of Lindernia ciliata. They are beta-sitosterol, stigmasterol and lup-20(29) en-3 beta-ol. Their structures were established by IR, MS, 1HNMR and 13CNMR spectroscopy. PMID- 8703352 TI - [Purification and properties of polysaccharide PPIII from Sterculia lychnophora Hance]. AB - Polysaccharide PP III was isolated and purified from aqueous extract of the seeds of Sterculia lychnophora. It is composed of galactose, arabinose and rhamnose, the molar ratio being 1 : 1.67 : 1.01 and molecular weight 162200. Rhamnoses in the main chain are linked by alpha-(1-->3) glycoside linkage. PP III C is its major fragment, and is composed of galactose and rhamnose with a molar ratio of 1 : 2. 78 and MW 62500. PMID- 8703353 TI - [A dynamic study on inhibitory effect of palustrine on mg2(+)-ATpase and Ca2(+) ATPase of brain synaptic vesicle membrane in rats]. PMID- 8703354 TI - [Effect of allitridum on macrophage-medicated cytotoxicity]. PMID- 8703355 TI - [Effect of ethanolic extract from root tuber of Polygonum multiflorum Thunb. on liver and brain monoamine oxidase in senescence-accelerated mice in vivo]. PMID- 8703356 TI - [Correlativity analysis of detoxicating effect of radix Glycyrrhizae on radix Aconiti preparata in sini decoction]. AB - Based on fixed dosages of Radix Aconiti Preparata and Rhizoma Zingiberis and six different dosages of Radix Glycyrrhizae, six different Sini Decoctions were prepared. The aconitine in these decoctions was determined by TLC-scanning. Statistical analysis was used in the experiment. It has been confirmed that the quantity of aconitine reduces with the increase of the quantity of Radix Glycyrrhizae, assuming a significant negative correlativity, r = -0.9945. PMID- 8703357 TI - [The status quo and prospect of experimental study on channel tropism of Chinese herbal drugs]. AB - The assay of metabolism and distribution of effective components of Chinese herbal drugs, microelement assay, pharmacodynamic assay and cAMP, cGMP assay are the essential methods usually used in the experimental study of channel tropism of Chinese herbal drugs. The problems of each method as well as the points for attention in the experimental study of channel tropism are discussed. PMID- 8703358 TI - [8 species of peroxidase isozymes and 2 varieties of Corydalis]. AB - The peroxidase isozymes of 8 species and 2 varieties of Corydalis were analysed by means of polyacrylamide gel electrophoresis. According to the isozymic zymograms, these plants may be divided into 3 groups. The taxonomy based on isozymic zymograms is in accord with that by morphology. The problem that the action of enzymes of Corydalls species is easily weakened or lost is also discussed. PMID- 8703359 TI - [Studies on Cnidium fruits by stereological method and computer image analysis]. AB - The fruits, pollen grains, vascular bundles, stomata and endosperm cells of 4 Cnidium species were studied and determined by stereological method and computer image analysis technique. Statistical data were presented. In the volume of fruit and the area of vascular bundle, Cnidium dahuricum is the biggest, then C. salinum, C. japonicum and C. monnieri successively. In the volume of pollen grain,C. salinum is the biggest, followed by C. dahuricum, C. japonicum and C. monnieri in succession. In the volume of endosperm cell, C. dahuricum is also the biggest, and then C. japonicum, C. salinum and C. monnieri in order. PMID- 8703360 TI - Exposure of postnatal rats to glucocorticoids suppresses the development of choline acetyltransferase-immunoreactive neurons: role of adrenal steroids in the development of forebrain cholinergic neurons. AB - Rat forebrain cholinergic neurons undergo dynamic developmental changes, showing a continuous increase in choline acetyl-transferase (ChAT) activity, during the early postnatal period. In adult rats, increases in circulating glucocorticoids result in decreases in activity of forebrain neuronal ChAT, thus raising the possibility that postnatal development of forebrain cholinergic neurons results from low levels of these hormones. In the rat, the first 2 weeks postnatally are characterized by very low levels of adrenal steroids. To understand the role of endogenous glucocorticoids in the development of forebrain cholinergic neurons, we studied the changes in ChAT immunoreactivity in forebrain cholinergic neurons of postnatal rats which had received daily subcutaneous injection of the synthetic glucocorticoid dexamethasone for 8 days. Immunohistochemical analysis of the rat pup forebrain revealed nearly complete obliteration of ChAT immunoreactive neurons in the caudate-putamen, especially in the dorsolateral region of the rostral part. At the same stage, treatment with dexamethasone induced significant decreases in both number and length of dendritic branches of ChAT-immunoreactive neurons in the substantia innominata and the diagonal band. Despite the marked alterations in the caudate-putamen and diagonal band, the ChAT immunoreactive neurons in other forebrain structures such as globus pallidus and medial septal nucleus showed little change. In the caudate-putamen, Nissl staining and specific labeling for nuclear DNA fragmentation exhibited no increase in number of dying cells following dexamethasone treatment, therefore indicating that the loss of ChAT immunoreactivity is not due to glucocorticoid induced cholinergic cell death. These observations demonstrated that the development of cholinergic neurons in rat pups was inhibited by prolonged glucocorticoid exposure, suggesting that low levels of adrenal steroids may promote the postnatal development of these neurons. PMID- 8703361 TI - The effect of intra-articular capsaicin on nerve fibres within the synovium of the rat knee joint. AB - The aim of this study was to establish the effects of intra-articular capsaicin (pelargonic acid vallinylamide) on synovial innervation of the rat knee. Rats were sacrificed 1, 2, 4 and 7 days after intra-articular injection of capsaicin and joint tissues stained with either conventional haematoxylin and eosin (H and E) or with specific antibodies to the calcitonin gene-related peptide (CGRP), substance P (both of which are markers for primary afferent fibres), the C flanking peptide of neuropeptide Y (CPON) (localised in postganglionic sympathetic fibres), or protein gene product 9.5 (a pan-neuronal marker). At lower concentrations (0.1% and 0.25%), capsaicin produced no change in peptide staining pattern or histological appearance. At 0.5% capsaicin, there was complete loss of nerve fibres showing positive staining for CGRP and substance P at all time points. Staining for CPON and protein gene product 9.5 was still present, but decreased, 1 and 2 days after treatment and virtually absent at 4 and 7 days. These findings provide evidence for partially selective denervation induced by 0.5% capsaicin, in contrast to 1% capsaicin which abolished staining for all peptide markers, indicating a total ablation of nerve fibres. A consistent but unexpected finding was the presence of a severe inflammatory response in joints treated with 0.5% and 1% capsaicin. An influx of polymorphonuclear leucocytes was found to occur within 4 h of injection, with progressive appearance of mononuclear cells after this time. We conclude that it is difficult to specifically deplete sensory nerve fibres from the synovium by means of local capsaicin injection. Although selective loss of staining for sensory nerve fibres could be achieved by injection of 0.5% capsaicin, there was progressive non-specific loss of post-ganglionic autonomic fibres which may be related to the severe inflammatory response provoked by the higher doses of capsaicin. PMID- 8703363 TI - Slice cultures of the imprinting-relevant forebrain area medio-rostral neostriatum/hyperstriatum ventrale of the domestic chick: immunocytochemical characterization of neurons containing Ca(2+)-binding proteins. AB - The forebrain area medio-rostral neostriatum/hyperstriatum ventrale, a presumed analogue to the mammalian prefrontal cortex, displays a variety of synaptic changes during auditory filial imprinting. In order to study the underlying basic mechanisms of this synaptic plasticity we developed slice cultures of the medio rostral neostriatum/hyperstriatum ventrale from newly hatched chicks. As a prerequisite for these investigations and in order to test the suitability of this system for future studies, we performed a thorough characterization of the in vitro tissue, of its cellular components and some of their biochemical features in comparison with in situ tissue. Since in situ the medio-rostral neostriatum/hyperstriatum ventrale has been previously shown to contain three distinct neuron populations characterized by the activity-regulated Ca(2+) binding proteins parvalbumin, calbindin D28K and calretinin, we used these proteins as neuronal markers to study the survival and preservation of the morphological features of medio-rostral neostriatum/hyperstriatum ventrale neurons in vitro. In agreement with in vivo studies the three Ca(2+)-binding proteins are confined to neuronal cells and they are not colocalized, i.e. they appear to characterize three different neuron populations. The immunoreactive neurons in medio-rostral neostriatum/hyperstriatum ventrale cultures to a certain extent appear to form synaptic contacts with each other, shown by the double immuncytochemical experiments. One difference between cells in vivo and in vitro is their soma size, which is much larger in vitro than in vivo. This and our previous study on neuronal morphology demonstrates that morphologically and biochemically intact neurons can be maintained in medio-rostral neostriatum/hyperstriatum ventrale slice cultures, which may thus provide a suitable in vitro system for further studies of neuronal and synaptic plasticity in vitro. PMID- 8703362 TI - A light and electron microscopic study of NADPH-diaphorase-, calretinin- and parvalbumin-containing neurons in the rat nucleus accumbens. AB - The rat nucleus accumbens contains medium-sized, spiny projection neurons and intrinsic, local circuit neurons, or interneurons. Sub-classes of interneurons, revealed by calretinin (CR) or parvalbumin (PV) immunoreactivity or reduced nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase histochemistry, were compared in the nucleus accumbens core, shell and rostral pole. CR, PV and NADPH-diaphorase-containing neurons are shown to form three non-co-localising populations in these three areas. No significant differences in neuronal population densities were found between the subterritories. NADPH-diaphorase containing neurons could be further separated morphologically into three sub groups, but CR- and PV-immunoreactive neurons form homogeneous populations. Ultrastructurally, NADPH-diaphorase-, CR- and PV-containing neurons in the nucleus accumbens all possess nuclear indentations. These are deeper and fewer in neurons immunoreactive for PV than in CR- and NADPH-diaphorase-containing neurons. CR-immunoreactive boutons form asymmetrical and symmetrical synaptic specialisations on spines, dendrites and somata, while PV-immunoreactive boutons make only symmetrical synaptic specialisations. Both CR- and PV-immunoreactive boutons form symmetrical synaptic specialisations with medium-sized spiny neurons and contact other CR- and PV-immunoreactive somata, respectively. A novel non carcinogenic substrate for the peroxidase reaction (Vector Slate Grey, SG) was found to be characteristically electron-dense and may be distinguishable from the diaminobenzidine reaction product. We conclude that the three markers used in this study are localised in distinct populations of nucleus accumbens interneurons. Our studies of their synaptic connections contribute to an increased understanding of the intrinsic circuitry of this area. PMID- 8703364 TI - Role of cyclo-oxygenase pathways in the stimulatory influence of immune challenge on the transcription of a specific CRF receptor subtype in the rat brain. AB - The aim of this study was to investigate the role of prostaglandins (PGs) on the expression of corticotropin-releasing factor (CRF) receptors in the brains of immune-challenged rats. Intravenous (i.v.) administration of indomethacin (0.8 mg/100 g body weight (b.w.)), an inhibitor of PG synthesis, was performed 15 min before the intraperitoneal (i.p.) injection of a high (250 micrograms/100 g b.w.), moderate (25 micrograms/100 g b.w.), or low (2.5 micrograms/100 g b.w.) dose of the immune activator lipopolysaccharide (LPS). Three and six hours after the i.p. treatment with the endotoxin LPS, male Sprague-Dawley rats (230-260 g) were sacrificed. Frozen brains were mounted on a microtome and cut from the olfactory bulb to the medulla in 30-microns coronal sections. mRNAs encoding CRF receptors (types 1, 2 alpha, and 2 beta) were assayed by in situ hybridization using 35S-labeled riboprobes. Strong basal levels of CRF1 receptor transcript were detected in multiple regions of the brain, whereas CRF2 alpha receptor message was highly localized in few structures of the limbic system and positive signal for CRF2 beta receptor mRNA was observed only in the choroid plexus. The transcription of the gene encoding the CRF type 1 (but not types 2) receptor was highly stimulated by LPS administration in selective hypothalamic nuclei. Indeed, a high dose of LPS caused strong expression of CRF1 receptor mRNA in both parvocellular and magnocellular paraventricular nucleus (PVN) and in the supraoptic nucleus (SON), although low and moderate doses of endotoxin induced a more specific expression of this transcript in the parvocellular division of the PVN. Pretreatment with indomethacin did not prevent the induction of CRF1 receptor transcription in the PVN of rats injected with a high dose of LPS. In contrast, inhibition of cyclo-oxygenase pathways significantly inhibited the expression of CRF1 receptor in the PVN and SON of rats sacrificed 6 h after being injected with a moderate or a low dose of LPS; the CRF1 receptor mRNA levels were approximately three (moderate dose) and two (low dose) times higher in rats receiving the endotoxin alone than in those submitted to a treatment combining both i.v. indomethacin and i.p. LPS. These results indicate that the mRNA encoding the type 1 but not the type 2 CRF receptor is specifically regulated in endocrine hypothalamus of immune-challenged rats, whereas the role of PGs in mediating the stimulatory influence of immune challenge on the transcription of CRF1 receptor in the PVN and SON seems to depend on the severity of this systemic stressful situation. PMID- 8703365 TI - New insight on immune tolerance from transgenic mouse models. PMID- 8703366 TI - Increased basal tone and hyperresponsiveness to acetylcholine and ergonovine in spasm related coronary arteries in patients with variant angina--basal coronary artery tone in patients with variant angina. AB - In patients with variant angina, previous data have been inconclusive as to whether basal coronary artery tone is elevated at the spastic and non-spastic sites. Thus, the purpose of this study was to assess the basal coronary artery tone and the responsiveness to acetylcholine (Ach) and ergonovine (Erg) in patients with variant angina. We compared the basal coronary artery tone and the constrictive responses to Ach and Erg between 31 patients (Group 1) with variant angina in whom spasm was provoked by the low doses of Ach (intracoronary 20 micrograms) or Erg(intravenous 50 micrograms) and 35 patients (Group 2) provoked by higher doses of Ach (intracoronary 100 micrograms) or Erg (intravenous cumulative dose of 350 micrograms), and 26 control subjects. Patients with variant angina in whom spasm was provoked by low doses of Ach or Erg, had a higher incidence of mixed disease, multi-vessel spasm and higher disease activity. The basal coronary artery tone at the spastic and nonspastic sites of spasm related artery was significantly more elevated in Group 1 than that in Group 2 (44 +/- 17 vs 14 +/- 11% and 26 +/- 14 vs 16 +/- 10% respectively, P < 0.05), but not in the nonspasm related artery, The magnitudes of vasoconstrictive responses to Ach and Erg at the nonspastic sites were also greater in Group 1 than those in Group 2 and the control groups (Ach; 40 +/- 20 vs 26 +/- 11, 27 +/- 12%: Erg; 37 +/- 18 vs 12 +/- 8, 13 +/- 10% respectively, P < 0.05). However, the basal coronary artery tone was not elevated at the spastic and nonspastic sites in Group 2 compared to the in control subjects. These findings suggest that the basal coronary artery tone is increased in patients with variant angina with higher disease activity at the spastic sites and nonspastic sites of the spasm related artery, and this may be related to the occurrence of coronary artery spasm. PMID- 8703367 TI - Efficacy of in vitro treatment of chronic myelogenous leukemia cell line, K562 cells, using 4-hydroperoxycyclophosphamide, alpha-interferon and gamma interferon. AB - Alpha-interferon (alpha-IFN) and gamma-interferon (gamma-IFN) are highly effective biologic agents in the treatment of chronic myelogenous leukemia (CML). 4-hydroperoxycyclophosphamide (4-HC) is the most commonly used chemotherapeutic drug when purging bone marrow contaminated with malignant cells. To investigate the efficacy of alpha-IFN, gamma-IFN and 4-HC as purging agents in CML patients, K562 cells, a human CML cell line, were treated in vitro with these drugs singly or in combination. The cytotoxic effect was evaluated by MTT assay. Data were analyzed for synergism by the median effect principle of Chou and Talalay. Using Southern blot analysis, the effect of purging on the suppression of bcr gene rearrangement was also evaluated. Alpha-IFN, gamma-IFN and 4-HC all showed good concentration-dependent cytotoxic effect. Median cytotoxic doses of alpha-IFN gamma-IFN and 4-HC were 0.03 microM 0.20 microM and 1.97 microM respectively, and molar ratios of median doses of alpha-IFN to 4-HC, gamma-IFN to 4-HC and alpha IFN to gamma-IFN were 1:80, 1:10 and 1:8, respectively. The combination treatment of alpha-IFN and 4-HC, and gamma-IFN and 4-HC showed significant synergistic cytotoxic activities, but combination of alpha-IFN and gamma-IFN was antagonistic. Status of bcr gene rearrangements in K562 cells was not changed after the treatments. Our results appear to provide an efficient method for removing CML cells from the bone marrow of CML patients. PMID- 8703368 TI - Expression of urokinase-type plasminogen activator, its receptor, and its inhibitor in gastric adenocarcinoma tissues. AB - The plasminogen and plasmin system, which is mainly regulated by urokinase-type plasminogen activator (uPA), its receptor (uPAR) and its inhibitor (PAI-1), is generally believed to play a role in cancer invasion and metastasis. This study was conducted to investigate the role of uPA, uPAR and PAI-1 in the invasion and metastasis of gastric adenocarcinoma. The expression of mRNAs for uPA and PAI-1 was determined by Northern blot analysis in nine primary gastric cancer tissues, nine paired metastatic lymph nodes and normal gastric mucosa. The mRNA of uPA was not or faintly detected in normal mucosa, while the expression was increased in both primary gastric cancer tissues and metastatic lymph nodes to a similar degree. The mRNA expression for PAI-1 in the gastric cancer tissues was not different from that in the paired metastatic lymph nodes and normal mucosae. uPAR was determined by immunohistochemical staining, demonstrating that five (56%) and six (67%) out of nine primary gastric cancer tissues and nine paired metastatic lymph nodes were positive, respectively and the intensity was stronger in metastatic lymph nodes. The results support the concept that most gastric cancer cells may have an innately moderate level of uPA and uPAR, and that increase of uPAR expression can be considered to be closely associated with cancer invasion and metastasis. PMID- 8703369 TI - A case-control study on the risk factors of hepatitis C virus infection among Koreans. AB - In order to identify the risk factors for hepatitis C virus (HCV) infections, a case-control study was conducted from September 1993 to April 1994. HCV infection was confirmed by the second generation of recombinant immunoblot assay. Sixty four cases and 128 controls matched for age and sex with a 1:2 ratio of cases to controls were enrolled. Exposure data were obtained from all participants by self administered questionnaire and the odds ratios of possible risk factors of HCV infection analysed. Sixty-four cases consisted of forty-two patients with chronic hepatitis, nine with cirrhosis, one with hepatocellular carcinoma, and twelve with normal liver function. History of acute hepatitis (OR 3.9) and transfusion (OR 2.4) were associated with an increased risk of HCV infection. Operation, acupuncture, endoscopy, tooth extraction, tattooing, ear piercing, needle sharing and family history of hepatitis were not associated with an increased risk of HCV infection. In conclusion, transfusion remains the major route of transmission of HCV in Korea. PMID- 8703370 TI - Immunohistochemical expression of phospholipase C in global and focal ischemic encephalopathy in gerbil: relationship with morphological changes. AB - Phospholipase C (PLC) and related enzymes in signal transduction system are closely linked to cellular damage in ischemic encephalopathy. This study was undertaken to elucidate the time sequential changes of PLC isoenzymes (beta and gamma) in vulnerable areas of hippocampus in global ischemia and infarcted area in focal infarction. Mongolian gerbils were used because of their susceptibility to ischemic encephalopathy and divided into the following groups: the bilateral ischemia with various reperfusion periods group, unilateral progressive ischemia group, and focal ischemia group induced by infusion of iron particles through the femoral artery. The changes of PLC isoenzymes were observed immunohistochemically and matched with morphological changes. In the global ischemia with reperfusion group, the changes were most significant in hippocampus. Sequential changes of neurons such as red neurons at an early stage progressed to pknotic neurons at a later stage were noted with typical delayed neuronal damage in the corns ammonis (CA) 1 subfield of hippocampus. Red neurons and pyknotic neurons as well as intracytoplasmic inclusion in 3 to 24 hours of reperfusion showed loss of PLC isoenzymes as well as tubulin. The changes of PLC expression were corresponding to the degeneration of neurons with no discernible time sequential changes in remaining neurons. In the unilateral hemispheric progressive ischemia group, ischemic damage was far more marked and extensive with no selective injury pattern according to time and location. At 1 day, there was diffuse vacuolization and necrosis of neuropil with a loss of neuron. Admixed surviving neurons and vacuolated neuropil showed increased reaction to anti-PLC antibodies, which could be either an evidence of protein synthesis responding to ischemic insult or an artifactual change. Focal ischemia group showed time sequential changes of blood vessels and white blood cells with necrosis of surrounding tissue. Degenerating hippocampal neurons in infarction also showed a strong positive reaction to anti PLC antibody, which was most likely due to condensation of cytoplasm rather than increased synthesis. This study showed different changes of PLC expression in global ischemic encephalopathy with reperfusion, progressive ischemia, and focal infarction, which suggested different pathophysiologic mechanism between these conditions. PMID- 8703371 TI - Relations among traumatic subdural lesions. AB - Acute subdural hematoma (ASDH), chronic subdural hematoma (CSDH) and subdural hygroma (SDG) occur in the subdural space, usually after trauma. We tried to find a certain relationship among these three traumatic subdural lesions in 436 consecutive patients. We included all subdural lesions regardless of whether they were main or not. We evaluated the distribution, age incidence and interval from injury to diagnosis of these lesions, and the frequency of new subdural lesions in each lesion. ASDH constituted 68.6%, SDG 15.8%, and CSDH 15.6%, Age incidence of CSDH was similar to that of SDG, but differed from that of ASDH. Mean interval from injury to diagnosis was 0.4 days in ASDH, 13.4 days in SDG, and 51.6 days in CSDH. Focal brain injuries accompanied in 37.5% of ASDH, 5.8% of SDG, and no CSDH. In ASDH, 2 recurrent ASDHs, 17 SDGs and 9 CSDHs occurred. In SDG, 3 postoperative ASDHs and 8 CSDHs occurred. In CSDH, 2 postoperative ASDHs, 2 SDGs and 1 CSDH occurred. These results suggest that the origin of CSDH is not only ASDH, but also SDG in upto a half of cases. SDG is produced as an epiphenomenon by separation of the dural border cell layer when the potential subdural space is sufficient. A half of CSDHs may originate from ASDHs. ASDH may occur in CSDH by either a repeated trauma or surgery. Such transformation or development of new lesions is a function of a premorbid condition and the dynamics between the absorption capacity and expansile force of the lesion. PMID- 8703372 TI - Ecthyma gangrenosum associated with aplastic anemia. AB - Ecthyma gangrenosum is a characteristic skin lesion of systemic infection due to Pseudomonas aeruginosa. It has a high incidence in patients with chronic disease and impaired defense mechanisms. Early diagnosis and appropriate systemic antibiotic therapy is crucial since its mortality rate is very high. We report a case of ecthyma gangrenosum in aplastic anemia. PMID- 8703373 TI - Unsuspected gastric granulocytic sarcoma in a patient with myelodysplastic syndrome. AB - Granulocytic sarcoma (GS) is an uncommon and localized extramedullary tumor composed of immature granulocytic cells. Most GS reported in large series were not associated with overt acute myelogenous leukemia. Gastric perforation occurred during prednisolone therapy in a 72-year-old Japanese male with a four month history of a myelofibrosis-like state. Subtotal gastrectomy was performed for a suspected gastric ulcer perforation. Gastric histologic, immunohistochemical and cytochemical examination revealed diffuse infiltration by sheets of myeloblasts and promyelocytes with scant or moderately abundant cytoplasm including a few eosinophilic myelocytes. Bone marrow study done in one month after the operation disclosed refractory anemia with excess of blasts (RAEB). Leukemic transformation occurred two months later, and a subcutaneous tumor appeared on the forehead. The forehead tumor predominantly consisted of myeloblasts without evidence of maturation. Both the stomach and forehead tumors were examined immunohistochemically with a panel of monoclonal antibodies (LCA, L26, MT1, UCHL1, OPD4, LN-1, LN-2, LN-3, MB1, Leu-M1, PM) and polyclonal antibodies (lysozyme, alpha 1-antitrypsin, alpha 1-antichymotrypsin, S-100 protein, lactoferrin), as well as naphthol-ASD-chloroacetate esterase staining to investigate and characterize the reliable marks for GS, and the patient was diagnosed as GS. We found that gastric GS may occur in a myelofibrosis-like state followed by RAEB of myelodysplastic syndrome and that naphthol-ASD-chloroacetate esterase staining and immunohistochemical detection of MT1, lysozyme, and alpha 1 antitrypsin were the most reliable markers for confirming the diagnosis of GS. PMID- 8703374 TI - Minimally differentiated acute myelogenous leukemia presented with multiple cervical lymphadenopathy. AB - Lymphadenopathy is a relatively uncommon finding of minimally differentiated acute myelogenous leukemia (AML-MO). We experienced a case of AML-MO in a 57-year old man initially presented with multiple cervical lymphadenopathy. Bone marrow aspiration revealed myeloblasts, which were negative for myeloperoxidase, Sudan black B, Periodic acid-Schiff, non-specific esterase and double esterase reaction. In cell surface marker studies, CD13, CD14, CD33, CD34, CD45 and HLA-DR were present. CT scan of neck demonstrated multiple lymphadenopathy at both internal jugular chains, spinal accessory chains and submandibular area. He died about two weeks after diagnosis without specific treatment. PMID- 8703375 TI - Selective inhibition of polymorphonuclear leukocytes by immunosuppressive concentration of prostaglandin E2. AB - Prostaglandin E2(PGE2) has been implicated as an immunosuppressive agent and plasma levels of PGE2 are elevated in patients sustaining thermal injury. We examined the effect of 10(-7) M prostaglandin E2(PGE2) on human polymorphonuclear leukocytes (PMN) to determine whether it directly inhibits stimulated responses of these cells. At this concentration, PGE2 alone was incapable of stimulating PMN intracellular hydrogen peroxide production (indirectly assayed by fluorescence of 2',7'-dichlorofluorescin) or expression of the PMN CD11b/CD16 surface glycoproteins. PMN incubated in the presence of the soluble stimuli phorbol myristate acetate(PMA, 100 ng/ml) or recombinant human C5a(rHC5a, 10(-8) M) generated significant amounts of hydrogen peroxide, increased their CD11b expression and decreased their CD16 expression. Pre-incubation of cells with PGE2 caused significant inhibition of all the observed changes stimulated by rHC5a. In contrast, events stimulated by PMA were not affected by preincubation of cells with PGE2. We conclude that PGE2, in concentrations identical to those found in the plasma of patients with burn injuries, is capable of selectively inhibiting some stimulated events and phenotypic expression of PMN in vitro study. PMID- 8703376 TI - Fibroepithelial ureteral polyp--a case report; endoscopic removal of large ureteral polyp. AB - We report a case of primary fibroepithelial polyp of the right midureter. The patient was a 41-year-old-woman, complaining of right flank pain. An excretory urogram revealed right hydronephrosis and a filling defect of the the right midureter. The filling defect was produced by a large fibroepithelial polyp that was diagnosed and removed by ureteroscopy without open surgery. Large fibroepithelial ureteral polyps are relatively rare and ureteroscopy is the gold standard of diagnosis for ureteral filling defect. PMID- 8703377 TI - Ovarian serous cystadenoma associated with Sertoli-Leydig cell tumor--a case report. AB - We Describe a case of ovarian serous cystadenoma having Sertoli-Leydig cell tumor, well differentiated, in the cystic septum. Well differentiated Sertoli Leydig cell tumor coexisting with other tumor, including serous tumor, has not yet been described. In all cases of Sertoli-Leydig cell tumor with heterologous components or other tumors, the androblastomatous components are intermediately or poorly differentiated. The present case revealed a well differentiated Sertoli Leydig cell tumor arising in a septum of serous cystadenoma, as a circumscribed nodule. With these findings, we discuss the possibility of this Sertoli-Leydig cell tumor, considered a mural nodule, which is well established in cystic common epithelial tumors of the ovary. PMID- 8703378 TI - Uterine endometrial stromal sarcoma with rhabdoid and smooth muscle differentiation. AB - Uterine and extrauterine tumors composed of cells featuring endometrial stromal cells often show ovarian sex cord-like structures and smooth muscle differentiation. A few cases of endometrial stromal tumors showing rhabdoid differentiation have been reported. The present case is a 20-year-old woman with endometrial stromal sarcoma that had sex cord-like structures, smooth muscle components and rhabdoid differentiation. PMID- 8703379 TI - Deletion of Rb1 gene in late osteosarcoma from survivor of unilateral retinoblastoma--a case report. AB - It has been well known that the survivors of retinoblastoma are prone to have osteosarcoma. But the secondary tumor usually occurs in bilateral, hereditary type of retinoblastoma. We report one case of osteosarcoma in a survivor of unilateral, sporadic retinoblastoma. A fourteen year old male presented with a painfully swollen distal forearm of 2 month duration. He had enucleated his left eye 10 years ago due to retinoblastoma with no other adjuvant therapy. We managed him with our conventional protocol and identified deletion of Rb gene from his pathological specimen by using the PCR-RFLP method. This result is unusual for unilateral nonhereditable retinoblastoma and may suggest gene level change even in sporadic cases. And Rb gene study may be helpful for unilateral, sporadic retinoblastoma patient in detecting the possibility of late osteosarcoma. PMID- 8703380 TI - Experimental strategies enabled by C-terminal protein sequence analysis. PMID- 8703381 TI - Selected risk factors in pediatric adenotonsillectomy. AB - OBJECTIVE: To evaluate the ability of a set of cost-effective criteria to identify before surgery the pediatric patients in whom perioperative respiratory compromise is most likely to develop after adenotonsillectomy. SETTING: A children's hospital medical center. DESIGN: Prospective study using preoperative parental questionnaires and perioperative respiratory status documentation. PATIENTS: All patients scheduled at the outpatient clinic were eligible. MAIN OUTCOME MEASURE: The development of respiratory compromise as defined by at least 1 of the following occurring more than 2 hours after surgery: an oxygen desaturation level of less than 90%, an obstructive breathing pattern, or respiratory distress requiring intervention. RESULTS: The risk of respiratory compromise was significantly increased in patients who were younger than 3 years (P < .001) and in those who had neuromuscular disorders (P < .05), chromosomal abnormalities (P < .005), difficulty in breathing during sleep (P < .005), restless sleep (P < .01), loud snoring with apnea (P < .05), or an upper respiratory tract infection within 4 weeks of surgery (P = .005). Respiratory compromise did not develop in any patients who did not snore (P < .05). CONCLUSIONS: A complete history that includes symptoms suggestive of sleep apnea will assist in the preoperative identification of pediatric patients most at risk for perioperative respiratory compromise after undergoing adenotonsillectomy. Such patients might benefit from overnight observation in a hospital setting. However, when snoring is absent, outpatient surgery is appropriate, as the risk of respiratory compromise is minimal. PMID- 8703382 TI - Nasal growth after external septoplasty in children. AB - OBJECTIVE: To assess the impact of external septoplasty surgery on nasal growth in children. DESIGN: Twelve anthropometric measurements (9 linear and 3 angular) were obtained in patients who previously underwent external septoplasty surgery for severe nasal obstruction caused by septal deviation anterior to the nasal spine. Surgery consisted of excision, refashioning, and reinsertion of the quadrilateral cartilage. From these 12 measurements, 5 proportional indexes were calculated, and then all measurements and proportions were compared with previously published norms. Follow-up measurements were taken at least 2 years after surgery (average, 3.4 years). SETTING: The Hospital for Sick Children, Toronto, Ontario, a tertiary care children's hospital. PARTICIPANTS: Twenty-eight patients who underwent external septoplasty surgery between the ages of 6 and 15 years. RESULTS: The principal measurements of the face and nose were within 1 SD of the normative mean for the majority of those in the study group. This was true for nasal height, nasal tip protrusion, nasal width, columella width, columella length, upper face height, face height, face width, inclination of the upper face, inclination of the nasal dorsum, and inclination of the columella. Values for 4 of the 5 proportional indexes were also overwhelmingly in the normal range. Twenty-nine percent of nasal dorsum measurements and 57% of nasal dorsum indexes were more than 2 SDs from the mean, indicating a predominance of short nasal dorsums. CONCLUSIONS: External septoplasty does not affect most aspects of nasal and facial growth, but it may negatively influence growth of the nasal dorsum. Prospective studies are needed to clarify this issue. PMID- 8703383 TI - Keratinocyte differentiation in acquired cholesteatoma and perforated tympanic membranes. AB - OBJECTIVE: To evaluate the type of differentiation of keratinocytes of acquired cholesteatoma and its significance for cholesteatoma invasiveness. DESIGN: Forty acquired cholesteatomas and 10 tympanic membranes with persisting perforations were snap frozen and processed for immunohistochemical studies. Cytokeratin antibodies that represented all subgroups and antibodies that were directed against collagen components of the basal lamina were applied. Expression of these constituents was scored by using light microscopy. RESULTS: The phenotype of the matrix was generally characterized by an extension of expression of basal cell cytokeratin 14 and hyperproliferation-associated cytokeratins 6, 16, and 17 into the suprabasal cell layers, while the expression of keratinization marker cytokeratin 10 was down-regulated. These features varied greatly at different sites of the matrix and were most marked at the advancing front of the cholesteatoma. A comparable expression pattern, but less pronounced, was observed at the epidermal front of the mucocutaneous junction of the tympanic membrane perforations. This phenomenon was invariably associated with a mononuclear cell infiltrate in the dermis at both junctions. The basal lamina was always intact. CONCLUSIONS: Acquired cholesteatomas show hyperproliferative features. There is a striking similarity between the pronounced expression of this phenotype and the associated inflammation at the mucocutaneous junctions of cholesteatomas and tympanic membrane perforations and those that are observed after epidermal injury. This indicates that epidermis and middle ear epithelium do not form stable junctions and the front can be considered to be a persisting epidermal defect. This involves the permanent presence of "activated keratinocytes" in the junction area that will lead to proliferation and migration, when additional triggers are present. PMID- 8703384 TI - Pseudomonas aeruginosa in otitis externa. A particular variety of the bacteria? AB - BACKGROUND: Pseudomonas aeruginosa rarely affects the epithelium in healthy persons except for the external ear canal, raising the possibility that P aeruginosa in otitis externa is a specific variety that displays particular characteristics. DESIGN: A cohort study was designed to outline distinct characteristics of P aeruginosa in otitis externa compared with P aeruginosa in other infections. The study period was October 1, 1994, to March 27, 1995. PATIENTS: Isolates of P aeruginosa from nonhospitalized patients were collected at the bacteriological laboratory at Karolinska Hospital, Stockholm, Sweden; there were 53 strains of P aeruginosa isolated from otitis externa and 59 strains of P aeruginosa from varicose ulcers and urinary tract infections. METHODS: Pseudomonas aeruginosa was characterized by pigmentation, growth habits, production of mucoid, and biochemical characteristics. RESULTS: Pseudomonas aeruginosa in otitis externa produced less pyocyanin and less urease and exhibited no mucoid-producing strains. CONCLUSIONS: Pseudomonas aeruginosa in otitis externa displayed fewer of the usual biochemical features of the species than did the strains isolated from other infections. Some of these features, such as the production of pyocyanin, are influenced by nutritional factors; strains found in otitis externa probably represent the type of strains present in the natural habitat in water, as opposed to the strains that have adapted to the environment of other human infections. Increased knowledge of the characteristics of the strains found in otitis externa is important in understanding the pathogenesis of the disease and why P aeruginosa is the dominant infectious agent in otitis externa. PMID- 8703386 TI - Conductive hearing loss following retrolabyrinthine surgery. AB - BACKGROUND: Conductive hearing loss can occur following neuro-otological procedures due to bony ossicular fixation resulting from viable bone dust remains. OBJECTIVE: To highlight the frequency and features of the complication of conductive hearing loss following retrolabyrinthine surgery. DESIGN: In a review of 77 retrolabyrinthine neuro-otological procedures, 3 cases (3.9%) of postoperative conductive hearing loss were encountered. RESULTS: Bone dust can cause a postoperative conductive hearing deficit that becomes apparent 6 months following surgery, and progressive deterioration can occur up to 18 months. The anatomical areas of ossicular fixation are the incudomalleal joint in the attic and around the stapes in the oval window niche. Excision of these bony accumulation particles does not revert the conductive hearing loss. CONCLUSIONS: We advocate the use of occlusive material in the aditus during temporal bone surgery to prevent bone dust accumulation in the attic and middle ear. In the event of such a complication, an incus transposition ossiculoplasty is recommended. PMID- 8703385 TI - A sensitive animal model to assess acute and chronic ototoxic effects. AB - BACKGROUND: Ototoxic side effects of aminoglycosides or cisplatin are strongly influenced by the nutritional state of the experimental animal. We previously demonstrated that guinea pigs fed a low-protein diet are a sensitive model to investigate chronic ototoxic effects of drugs. We now demonstrate that this model is equally sensitive to acute ototoxic insults. SUBJECTS AND DESIGN: Guinea pigs were fed either a full-protein (18.5% protein) or a low-protein (7% protein) diet. Gentamicin sulfate was given as a single injection (100 mg/kg of body weight subcutaneously), followed 90 minutes later by varying concentrations of ethacrynic acid intravenously. Auditory function was assessed by auditory brain stem response and morphologic characteristics in surface preparations of the organ of Corti. RESULTS: In animals fed a full-protein diet, 35 mg of ethacrynic acid per kilogram of body weight was required for a significant threshold shift at all frequencies tested (3, 8, and 18 kHz). In animals fed a low-protein diet, 15 mg of ethacrynic acid per kilogram significantly elevated threshold at all frequencies, and 20 mg/kg caused a profound threshold shift (> 90 dB). Hair cell loss in the organ of Corti paralleled the pathophysiological findings. CONCLUSIONS: A metabolically challenged animal may better represent the situation of a severely ill patient undergoing drug therapy. Therefore, studies in diet restricted animals may be more appropriated than studies in healthy animals for an assessment of potential adverse therapeutic side effects. PMID- 8703387 TI - Absent vestibular evoked myogenic potentials in vestibular neurolabyrinthitis. An indicator of inferior vestibular nerve involvement? AB - BACKGROUND: Benign paroxysmal positioning vertigo (BPPV) is generally thought to be caused by canalolithiasis in the posterior semicircular canal, an organ that is innervated by the inferior vestibular nerve. We hypothesized that absent vestibular evoked myogenic potentials (VEMPs) would indicate involvement of the inferior vestibular nerve and that posterior semicircular canal-type BPPV could not develop after vestibular neurolabyrinthitis (VNL) in patients with absent VEMPs. OBJECTIVE: To find out if VEMPs could be helpful in evaluating involvement of the inferior vestibular nerve in acute VNL. DESIGN: We reviewed the VEMP findings in 47 patients (34 men and 13 women) with acute VNL, 10 of whom had then developed posterior semicircular canal-type BPPV. RESULTS: While p13-n23, the first positive-negative peak of the VEMP, was ipsilaterally present on stimulation of the unaffected side in all patients, it was absent on the affected side in 16 patients (34%). The absence or presence of p13-n23 was independent of the results of caloric tests, pure tone audiometry, and auditory brain-stem responses. Typical posterior semicircular canal BPPV developed in 10 of the 47 patients after the acute attack of VNL, always on the same side as the neurolabyrinthitis. The p13-n23 potentials were preserved on stimulation of the affected ear in all 10 patients with BPPV. CONCLUSIONS: These results suggest that if VEMPs are absent from an ear that has suffered acute VNL, then posterior semicircular canal BPPV is unlikely to develop as a consequence of the VNL. The reason for this appears to be that the absence of VEMPs is due to involvement of the inferior vestibular nerve or involvement of the structures that it innervates. PMID- 8703388 TI - Plasma endothelin level in the acute stage of Bell palsy. AB - OBJECTIVE: To determine the plasma endothelin level in the acute stage in patients with Bell palsy (based on the hypothesis that endothelin, which is a potent vasoconstrictor, may play a role in the mechanism of the onset of facial nerve paralysis and in view of the fact that the etiology of Bell palsy is still a maze of unknowns). DESIGN: The study involved 62 patients with the acute stage (tested within 10 days of onset) of Bell palsy (i.e., idiopathic acute peripheral facial paralysis) and an additional 36 healthy persons who served as control subjects. To determine the content of endothelin, 2 ml of plasma samples was collected from each subject. Endothelin-1 was extracted and analyzed by a radioimmunoassay by using anti-endothelin-1 antibody. SETTING: Nihon University Itabashi Hospital, a referral and institutional center in Tokyo, Japan. RESULTS: The patients who were suffering from Bell palsy exhibited a statistically significant (P < .01) increase in the endothelin level compared with that in the 36 normal control subjects. An age-matched comparison (ranges, 20-29 years and 30 39 years) of patients with Bell palsy with normal control subjects revealed a significant difference between the normal group and the group with Bell palsy in the plasma endothelin level for both age groups that were tested (P < .01). The mean value of the endothelin level in patients with Bell palsy was maximal on day 5, and the percentage of patients with abnormally elevated endothelin levels was 100% from days 6 to 9. CONCLUSION: Endothelin, which has potent vasoconstrictive effects, may contribute to the pathogenesis of the microcirculatory impairment that occurs in patients with Bell palsy, mainly by promoting secondary ischemia. PMID- 8703389 TI - A novel organ preservation protocol for advanced carcinoma of the larynx and pharynx. AB - OBJECTIVE: To pilot a targeted chemoradiation protocol for patients with advanced carcinoma of the larynx and pharynx that would circumvent upper aerodigestive tract dysfunction related to major oncologic surgery. DESIGN: Weekly intra arterial infusions of supradose cisplatin (150 mg/m2 per week x 4) rapidly delivered to the tumor bulk, simultaneous intravenous sodium thiosulfate for systemic drug neutralization, and conventional external-beam irradiation (1.80 2.00 Gy per fraction x 35) were used. Between February 1991 and April 1994, 42 patients were treated who would otherwise have required a major resection of the tongue base, pharyngeal wall, or larynx. MAIN OUTCOME MEASURES: Tumor response, toxic effects, disease control above the clavicle, preservation of the larynx, maintenance of oral nutrition, and overall and disease-related 2-year survival. RESULTS: Three complications were related to the weekly transfemoral superselective intra-arterial procedures performed 160 times. Grade 3 to 4 chemotoxic effects were infrequent, occurring in 9 (5.5%) of 160 cycles, and only 1 patient required a radiotherapy break because of severe mucositis. A complete response in the primary site was obtained in 36 (86%) of 42 patients, 2 of whom had residual disease in the neck. Median follow-up was 13 months (range, 3-46 months). To date, there have been 5 recurrences: 2 regional and 3 distant. The 2 year overall and disease-related survival was 64% and 76%, respectively. The rate of disease control above the clavicle at 2 years was 86%. CONCLUSIONS: We believe this chemoradiation protocol represents an effective management scheme for patients with advanced head and neck cancer while minimizing dysfunction and possibly improving survival. PMID- 8703390 TI - Assessing vocal function after chemoradiation for advanced laryngeal carcinoma. AB - OBJECTIVE: To identify objective parameters that could serve as valid indicators of vocal function in evaluating patients treated at multiple centers for head and neck cancer. DESIGN: Objective measures of voice were validated against functional outcome measures of voice and communication. SETTING: A multidisciplinary university-based ambulatory head and neck cancer clinic. SUBJECTS: Fifteen patients at varying posttreatment intervals after chemoradiation therapy for head and neck cancer and 5 volunteers with no cancer but with a history of heavy smoking and drinking. MAIN OUTCOME MEASURES: Acoustic and aerodynamic parameters were measured in patients with head and neck cancer after treatment with combined chemotherapy and irradiation and in control subjects. Communicative function was assessed by perceptual assessment of recorded voice samples, by a clinician-based communication score, and by patient self-rating of voice and communication. RESULTS: Both perceptual analysis and communication score were concordant with patient self-rating of voice and communication ability. Acoustic measures were abnormal in patients, and jitter scores correlated with listener ratings of perceived voice quality. However, acoustic measures did not relate to communication function, as assessed by either the patient or the clinician. Laryngeal resistance demonstrated a significant correlation with both patient and observer assessments of voice quality and communication function. Laryngeal resistance was on average twice as high in patients who complained of voice fatigue, but this difference was not statistically significant. CONCLUSIONS: Laryngeal resistance, determined during standardized phonation, is a reliable objective parameter of vocal function after chemoradiation therapy for head and neck cancer. A standardized clinician-based communication scale holds promise as a useful tool. Acoustic measures reflect the sound of the voice but not the ability to communicate effectively. PMID- 8703391 TI - Tumor angiogenesis as a prognostic factor in early oral tongue cancer. AB - OBJECTIVE: To evaluate the relationship between tumor angiogenesis and the subsequent development of cervical node metastases in patients with stage 1 carcinoma of the oral tongue. METHODS: The study group consisted of 25 patients with T1 carcinoma with no evidence of neck disease at presentation. Paraffin blocks from 10 patients, in whom neck node metastases were found shortly after surgery, were stained for factor VIII and compared with those from 10 randomly chosen patients in whom no evidence of recurrent disease was observed during prolonged followup. Microvessels were counted in each group. RESULTS: The microvessel counts correlated with metastatic disease. The mean +/- SD count was 103.41 +/- 40.72 per X 250 field in the patients with positive lymph node metastases and 50.03 +/- 13.74 per X 250 field in those with no lymph node metastases during follow-up (P < or = .05). CONCLUSIONS: The number of microvessels per X 250 field in the area of most intensive neovascularization in early oral tongue carcinoma may be an independent predictor of nodal metastases. Assessment of tumor angiogenesis may therefore prove valuable in selecting patients with early oral tongue carcinoma for aggressive therapy. PMID- 8703392 TI - Middle turbinate stabilization technique in endoscopic sinus surgery. AB - BACKGROUND: Lateralization of the middle turbinate with scarring and obstruction of the middle meatus after endoscopic ethmoidectomy has accounted for a high percentage of postoperative complications. OBJECTIVE: To evaluate a suture stabilization technique of the middle turbinates in an attempt to solve this problem and preserve the middle turbinate. DESIGN: Retrospective chart review of 31 consecutive patients undergoing endoscopic sinus surgery with suture stabilization of the middle turbinate. RESULTS: With the technique of suture stabilization of the middle turbinate in 60 operated-on sides of 31 patients, 59 sides showed the middle meatus to be patent without synechia or maxillary sinus ostium obstruction postoperatively. CONCLUSION: The complication of lateralization of the middle turbinate with scarring and obstruction of the middle meatus after endoscopic ethmoidectomy can be prevented with suture stabilization of the middle turbinate, and the middle turbinate can be preserved. PMID- 8703393 TI - Comparison of N-octyl-cyanoacrylate vs suture in the stabilization of cartilage grafts. AB - OBJECTIVE: To compare N-octyl-cyanoacrylate and suture control to determine the efficacy of this tissue glue preparation in securing grafted cartilage with an acceptable inflammatory reaction. DESIGN: Four 5-mm circular auricular cartilage grafts were harvested, and 2 of these grafts were either glued together with N octyl-cyanoacrylate or sewn together with nylon suture. These sandwich grafts were then glued or sewn to the periosteum of the calvaria. Histologic analysis for inflammation, necrosis, fibrosis, and foreign-body giant cell reaction was performed 2 and 6 weeks after implantation. We also measured cartilage graft migration from the original site of surgical implantation. SUBJECTS: Sixteen New Zealand white rabbits. The 2-week study group consisted of 7 rabbits (1 died of dehydration), and the 6-week group totaled 8. INTERVENTION: Each rabbit underwent auricular cartilage harvest and cartilage graft implantation to the parietal skull periosteum. RESULTS: We found no histological differences between the glue specimens and the suture controls. Similarly, our examination of graft migration yielded no differences between groups. CONCLUSIONS: N-octyl-cyanoacrylate was superior to standard suture technique by virtue of its speed, ease, and accuracy of application. No adverse inflammation was observed. Further studies, including clinical trials, are needed to assess the safety of N-octyl-cyanoacrylate in humans. PMID- 8703394 TI - Correction of velopharyngeal insufficiency with furlow palatoplasty. AB - OBJECTIVE: To evaluate the authors' experience with Furlow palatoplasty for velopharyngeal insufficiency. DESIGN: Review of 8 cases over a 3-year period. SETTING: Regional private practice hospital affiliated with a teaching university. PATIENTS: Eight patients aged 4 to 14 years with symptoms and signs of velopharyngeal insufficiency underwent Furlow palatoplasty. Four patients had submucous or soft palate clefts without previous intervention. Four patients had undergone previous surgery for cleft palate. INTERVENTION: All patients underwent Furlow palatoplasty. MAIN OUTCOME MEASURES: Fiberoptic nasopharyngeal examination and speech analysis. RESULTS: Seven of 8 patients exhibited good palate closure and marked improvement in speech. One patient exhibited good closure but had mild nasal emission and hypernasality, which responded to speech therapy after surgery. CONCLUSION: Furlow palatoplasty is a useful technique for the correction of velopharyngeal insufficiency in selected patients. PMID- 8703395 TI - Role of ischemic gradient in neovascularization of interpolated skin flaps. AB - OBJECTIVES: To investigate the significance of the ischemic gradient between interpolated skin flaps and the recipient bed and to determine its role in flap viability. DESIGN: Bilateral interpolated skin flaps were elevated in 10 pigs and intravenous fluorescein was used to define a border of pedicle perfusion. Flaps were assigned to 1 of 2 groups: proximal flaps, which were contained within the fluorescein border, or distal flaps, which extended 3 cm beyond the fluorescein border. Flaps were then transferred to an adjacent cutaneous defect and pedicles were divided after 2 weeks. The flap and pedicle portions were evaluated separately and viability was quantified. SUBJECTS: Ten adolescent pigs with bilateral flaps. None were withdrawn. INTERVENTION: Intravenous fluorescein (20 mg/kg) to determine extent of perfusion in the flaps. RESULTS: Proximal flaps (n = 10) maintained excellent viability with the pedicles attached. After pedicle division, however, partial flap and pedicle necrosis developed. Distal flaps (n = 10) promptly showed signs of ischemia and congestion but soon showed improved vascularity. A statistically significant improvement was seen in mean percent viability of the distal group compared with the proximal group (P < .05) (Student t test). CONCLUSION: The more ischemic flap-pedicle complex resulted in greater viability after pedicle division. The ischemic gradient seemed to represent a potent angiogenic stimulus to the recipient bed. PMID- 8703396 TI - Pathologic quiz case 1. Kimura disease. PMID- 8703397 TI - Pathologic quiz case 2. Cystic lymphangioma. PMID- 8703398 TI - Short-stay outpatient tonsillectomy. PMID- 8703399 TI - Short-stay outpatient tonsillectomy. PMID- 8703400 TI - Drainage after thyroid and parathyroid surgery. PMID- 8703401 TI - Long-term nasal mucosal tissue expansion use. PMID- 8703402 TI - Pseudomonas ear infections. PMID- 8703403 TI - Acquired laryngomalacia. PMID- 8703404 TI - Cellular adhesiveness, contractility, and traction: stick, grip, and slip control. AB - Translocation of cells over solid substrata depends on generation of motive force, in crawling tissue cells, brought about by regulated contractility of intracellular actomyosin. Intracellular contractile machinery has a direct, structural connection to the cell surface. Hence, regulated adhesiveness of the cell surface provides a mechanism whereby a cell can fine tune the extent of tractional forces that are necessary for effective translocation. Cells are able to control adhesiveness of surfaces (stick), contractility (grip), and the extent of traction exerted on the substratum (slip). Here, I discuss several aspects of local (subcellular) regulation of adhesiveness and contractility and speculate on how cells, given a choice of the substratum, decide on how and where to apply traction. PMID- 8703405 TI - Biological specificity and measurable physical properties of cell surface receptors and their possible role in signal transduction through the cytoskeleton. AB - It is proposed that the binding specificities of cell adhesion molecules are manifested in their measurable physical properties. A method specifically designed to measure the interfacial tension of cell aggregates is described. With the introduction of a statistical mechanical model, the measured values of tensions for aggregates consisting of genetically engineered cells with controlled adhesive properties are used to obtain information on the strength of individual receptor-ligand bonds. The strength of binding must depend on the receptor and its ligand and reflects the amino acid sequence of the binding proteins. Many of the cell surface receptors, being transmembrane proteins, are attached to the various macromolecular networks of the cytoskeleton; therefore, it is suggested that their ligation and ensuing conformational change may substantially affect the mechanical state of the cytoskeletal assemblies. Since these assemblies are believed to actively participate in intracellular signaling by transmitting signals from the cell membrane into the nucleus, the cell adhesion molecules may influence signaling in a predictable way through their measurable physical characteristics. In particular, varying bond strength at the cell surface may lead to differential gene regulation. PMID- 8703406 TI - Probing transmembrane mechanical coupling and cytomechanics using magnetic twisting cytometry. AB - We recently developed a magnetic twisting cytometry technique that allows us to apply controlled mechanical stresses to specific cell surface receptors using ligand-coated ferromagnetic microbeads and to simultaneously measure the mechanical response in living cells. Using this technique, we have previously shown the following: (i) beta 1 integrin receptors mediate mechanical force transfer across the cell surface and to the cytoskeleton, whereas other transmembrane receptors (e.g., scavenger receptors) do not; (ii) cytoskeletal stiffness increases in direct proportion to the level of stress applied to integrins; and (iii) the slope of this linear stiffening response differs depending on the shape of the cell. We now show that different integrins (beta 1, alpha V beta 3, alpha V, alpha 5, alpha 2) and other transmembrane receptors (scavenger receptor, platelet endothelial cell adhesion molecule) differ in their ability to mediate force transfer across the cell surface. In addition, the linear stiffening behavior previously observed in endothelial cells was found to be shared by other cell types. Finally, we demonstrate that dynamic changes in cell shape that occur during both cell spreading and retraction are accompanied by coordinate changes in cytoskeletal stiffness. Taken together, these results suggest that the magnetic twisting cytometry technique may be a powerful and versatile tool for studies analyzing the molecular basis of transmembrane mechanical coupling to the cytoskeleton as well as dynamic relations between changes in cytoskeletal structure and alterations in cell form and function. PMID- 8703407 TI - Mechanical basis of cell shape: investigations with the scanning acoustic microscope. AB - The shape of cells during interphase in sparse cultures often resembles that of fried eggs. XTH-2 cells, which have been derived from tadpole heart endothelia, provide a typical example of this type of shape. To understand the physical basis of this shape, the cytoskeleton of these cells has been investigated in detail. Subcellular elasticity data have been achieved by scanning acoustic microscopy (SAM). Their changes were observed during treatment of the cells with microtubule disrupting agents (colcemid and low temperature), and shape generation in giant cells produced by electro-fusion was observed with SAM, revealing the role of the nucleus as a force centering organelle. From these observations combined with well-documented observations on cellular dynamics described in the literature, a model is developed explaining the fried-egg shape of cells by means of interacting forces and fluxes (cortical flow, bulk flow of cytoplasm, microtubule mediated transport of cytoplasm) of cytoplasm. The model also allows the comprehension of the increase of tension in cells treated with colcemid. PMID- 8703408 TI - Mechanoreception at the cellular level: the detection, interpretation, and diversity of responses to mechanical signals. AB - Cells from diverse tissues detect mechanical load signals by similar mechanisms but respond differently. The diversity of responses reflects the genotype of the cell and the mechanical demands of the resident tissue. We hypothesize that cells maintain a basal equilibrium stress state that is a function of the number and quality of focal adhesions, the polymerization state of the cytoskeleton, and the amount of extrinsic, applied mechanical deformation. A load stimulus detected by a mechano-electrochemical sensory system, including mechanically sensitive ion channels, integrin-cytoskeleton machinery, and (or) a load-conformation sensitive receptor or nonreceptor tyrosine kinase, may activate G proteins, induce second messengers, and activate an RPTK or JAK/STAT kinase cascade to elicit a response. We propose the terms autobaric to describe a self-loading process, whereby a cell increases its stress state by contracting and applying a mechanical load to itself, and parabaric, whereby a cell applies a load to an adjacent cell by direct contact or through the matrix. We predict that the setpoint for maintaining this basal stress state is affected by continuity of incoming mechanical signals as deformations that activate signalling pathways. A displacement of the cytoskeletal machinery may result in a conformational change in a kinase that results in autophosphorylation and cascade initiation. pp60Src is such a kinase and is part of a mechanosensory protein complex linking integrins with the cytoskeleton. Cyclic mechanical load induces rapid Src phosphorylation. Regulation of the extent of kinase activation in the pathway(s) may be controlled by modulators such as G proteins, kinase phosphorylation and activation, and kinase inhibitors or phosphatases. Intervention at the point of ras-raf interaction may be particularly important as a restriction point. PMID- 8703409 TI - Type I and type VIII adenylyl cyclases constitute a family whose activation is coupled to cellular deformation through the action of calcium-calmodulin. AB - In certain tissues and cells, increases in concentrations of the second messenger cAMP are seen in response to mechanical or deformational stimuli. Type I and type VIII adenylyl cyclases, representing members of a family of calcium-calmodulin stimulated adenylyl cyclases, and type VII adenylyl cyclase were each stably expressed in human embryonal kidney (HEK) 293 cells. HEK 293 cells exogenously expressing either type I adenylyl cyclase or any one of three type VIII adenylyl cyclase splice variants respond to swelling with increases in cAMP, requiring the presence of calcium in the extracellular medium for such responsiveness. Type VII expressing HEK 293 cells failed to respond to swelling with increased cAMP but demonstrated potentiation of isoproterenol-stimulated activity. This is characteristic of the influence of protein kinase C on the activity of the type VII protein. The relative swelling responsiveness of HEK 293 cells expressing splice variants of the type VIII adenylyl cyclase is consistent with the relative EC50 values for calcium-calmodulin stimulation of these splice variants. This is consistent with the involvement of calmodulin and the requirement for increases in intracellular calcium in mediating swelling-induced acceleration of type VIII adenylyl cyclase activity. PMID- 8703410 TI - Simian virus 40 transformation alters the actin cytoskeleton, expression of matrix metalloproteinases and inhibitors of metalloproteinases, and invasive behavior of normal and ataxia-telangiectasia human skin fibroblasts. AB - Alterations in the actin cytoskeleton of normal cells result in changes in cell shape and adhesiveness and induce expression of matrix-degrading matrix metalloproteinases. We examined the effect of simian virus 40 transformation of normal and ataxia-telangiectasia human skin fibroblasts, a process that produces actin reorganization, altered cell morphology, and altered cell behavior, on expression of genes of the matrix metalloproteinase and tissue inhibitor of metalloproteinases gene families. Simian virus 40 transformation induced collagenase-1 gene expression; in contrast, stromelysin-1, 72-kDa gelatinase (gelatinase A), tissue inhibitor of metalloproteinases-1, and tissue inhibitor of metalloproteinases-2 genes were repressed. Transformation also altered the response of the fibroblasts to 12-O-tetradecanoylphorbol-13-acetate. Collagenase mRNA was induced in 12-O-tetradecanoylphorbol-13-acetate treated transformed cells up to 50-fold more than in untreated transformed cells or in 12-O tetradecanoylphorbol-13-acetate treated untransformed parent cells. In contrast, 12-O-tetradecanoylphorbol-13-acetate did not overcome the attenuated expression of stromelysin-1 in the simian virus 40 transformants. In addition, 92-kDa gelatinase (gelatinase B) was induced by 12-O-tetradecanoylphorbol-13-acetate only in the simian virus 40 transformants. The responses of gelatinase A and tissue inhibitor of metalloproteinases-1 to 12-O-tetradecanoylphorbol-13-acetate were unchanged. The pattern of altered proteinase expression after transformation was accompanied by a phenotypic alteration in cell invasion. The simian virus 40 transformants exhibited enhanced invasiveness through a basement-membrane-like matrix. These data demonstrate that enhanced invasiveness in simian virus 40 transformed cells is accompanied by changes in actin organization and expression of proteinases and inhibitors, as well as in the balance between proteinases and inhibitors in favor of proteinases. PMID- 8703412 TI - Modeling of molecular mechanisms of cell adhesion. AB - Cellular adhesion is a process of great importance in biology. We present a simple model of the adhesion process in which the molecular mechanisms involve a receptor, a ligand, and the cytoskeleton of the cell. Based on the energetic consideration of the process, we propose a molecular interpretation of the existing experimental data. The model suggests that the interaction of the receptor and (or) receptor-ligand with the cytoskeleton can have important influence on the formation and strength of the adhesion complex as well as on the subsequent interaction with different ligands. When conformational changes take place during the adhesion process, the characterization of the adhesion bonds based on chemical kinetics alone seems to be incomplete and must be supplemented by parameters, describing the functionality of the complex, i.e., change of the affinity for different ligands, as in the signal transduction, or the strength of the bond, as in the adhesion process. PMID- 8703411 TI - Dynamic reciprocity revisited: a continuous, bidirectional flow of information between cells and the extracellular matrix regulates mammary epithelial cell function. AB - Interactions between cells and the extracellular matrix (ECM) generate two classes of signals, mechanical and biochemical. In the case of the mammary epithelial cell, both are required to initiate ECM-dependent expression of the abundant milk protein beta-casein. Mechanical signals induce a cellular rounding, while functional biochemical signals are associated with an increase in tyrosine phosphorylation. These individual components are part of a complex signalling hierarchy that leads to the emergence of the fully functional lactational phenotype. Interestingly, both the assembly and disassembly of this hierarchy, which occur cyclically in vivo, are constantly modulated by dynamic and reciprocal interactions that take place within a functional unit composed of both the cell and the ECM. PMID- 8703413 TI - Influence of surface charges on cell adhesion: difference between static and dynamic conditions. AB - We tested the hypothesis that nonspecific repulsion, as a result of electrostatic forces and (or) steric stabilization effects, impaired adhesion more efficiently under dynamic than under static conditions. Cells from the human monocytic line THP1 were plated on a glass surface. Spherical particles bearing monoclonal antibodies specific for antigens expressed by THP1 cells (CD11b, CD18, CD35, CD64) were then added and adhesion was quantified. The effect of neuraminidase treatment of THP1 cells was also studied. Adhesion was then measured in a flow chamber under low shear flow (wall shear rate was 11 or 22 s-1), allowing a quantitative determination of cell adhesion frequency. The following conclusions were obtained: (i) under static conditions, neuraminidase treatment had little effect on adhesion (only CD18-mediated interaction was significantly increased at 4 degrees C after enzyme treatment); (ii) under dynamic conditions, neuraminidase treatment significantly increased binding; (iii) surprisingly, there was no clear relationship between the length of adhesion molecules involved in the interaction and binding efficiency; and (iv) such parameters as cell shape and topographical distribution of adhesion molecules may strongly influence adhesion under flow. It is concluded that a dynamic reorganization of the pericellular matrix following intercellular contact may play an important role in regulating adhesion. PMID- 8703414 TI - Modeling chemoattractant-elicited relocalization of myosin filaments in Dictyostelium. AB - Dictyostelium myosin is able to assemble into filaments that, when visualized under normal conditions, appear to be uniformly distributed throughout the cytoplasm. After stimulation by the chemoattractant cAMP, these filaments quickly diminish in the cellular medulla and accumulate in the cortex. A general hypothesis to explain the mechanism of this relocalization proposes that one or more of the chemical coefficients governing filament polymerization is precisely regulated by some sort of intracellular second messenger. To investigate this hypothesis we formulated a simple theoretical model of myosin polymerization and then used this model to analyze the resting state of the cell and various scenarios for initializing a transition to the activated state. In general, we found that the relocalization of filaments could be realized if a second messenger increased the elongation and (or) the nucleation coefficients for filament assembly in cortical ectoplasm and (or) if the messenger decreased these parameters in the cellular medulla. By comparing these limiting cases with experimental observations, we concluded that models in which redistribution of myosin is achieved by decreasing filament stability in the medulla are the most likely candidates. PMID- 8703415 TI - A new model for the research into rhythmic contraction activity of cardiomyocytes in vitro. AB - Heart cells continue to contract rhythmically after isolation and in culture in vitro. We describe a model of heart preparation in vitro that permits quantitative research on the frequency of contractions of cardiomyocytes. The chick embryo heart explants placed on a network of elastic glass fibers continued beating for months, recorded and analyzed with the methods of computer-assisted image analysis. The efficacy of this experimental model for the screening of effects of various agents on the frequency of contractions was examined by following the effects of nifedipine, caffeine, ethanol, and benzamide. The reversibility of the effects and the reproducibility of results were demonstrated quantitatively. The significance of a mechanical elastic load provided by glass fibers for the preservation of long-lasting contractile activity of cardiomyocytes is discussed and the common occurrence of oscillatory contraction processes in various eucaryotic cells is noted. PMID- 8703416 TI - Patterns of spontaneous motility in videomicrographs of human epidermal keratinocytes (HEK). AB - The subject of our observations was the spontaneous behaviour of normal and transfected human epidermal keratinocytes. Cell movements were recorded on video micrographs and analyzed by a mathematical approach, using new methods of image processing and statistical correlation analysis. Protrusive activity of single lamellae was examined using one-dimensional analysis of phase-contrast image sequences along section lines transversal to the cell edge. This method revealed high periodicity and correlation in the motility patterns of lamellae and ruffles. Two-dimensional correlation analysis of automatically digitized cell outlines was applied to detect spatiotemporal patterns and coordination of lamellar extension and retraction. Most cells showed regularly alternating pulsations of lamellar protrusions. In some extreme cases, extension waves rotating around the cell periphery were observed. The results were compared with computer simulations of two simple models for lamellar dynamics and shape deformation, based on few assumptions about chemical kinetics of F-actin and cytomechanical properties of the actin network, neglecting regulatory effects of actin-associated proteins or extracellular stimulations. The simulation results reproduced the main dynamical features of the observed real cells, indicating the possibility that the basic universal mechanism for lateral coordination of lamellipodial protrusion is the interplay between hydrostatic pressure and viscocontractile tension in the cortical F-actin-plasma membrane complex. PMID- 8703417 TI - Characterization of endothelial cell locomotion using a Markov chain model. AB - A Markov chain model was developed to characterize the two-dimensional locomotion of bovine pulmonary artery endothelial (BPAE) cells cultured with or without basic fibroblast growth factor (bFGF). This model provides a detailed description of the migration process by computing the following locomotory parameters: (i) the speed of cell locomotion; (ii) the expected duration of cell movement in any given direction; (iii) the probability distribution of turn angles that will decide the next direction of cell movement; (iv) the frequency of cell stops; and (v) the duration of cell stops. Eight directional states and a stationary state were used in our Markov analysis. From cell trajectory data, the transition probabilities among the various states and the waiting times for the directional and the stationary states were computed. The steady-state probabilities were also calculated to obtain the ultimate direction of cell motion and, thus, determine whether cell motion was random. Our results showed how the addition of bFGF enhanced the locomotory capability of BPAE cells. Cells cultured with 30 ng/mL bFGF had lower probability of moving to the stationary state than those cultured without bFGF. In addition, cells cultured with 30 ng/mL bFGF remained in the stationary state for shorter periods of time than cells cultured without bFGF. In both these cases, however, the transition probabilities from the stationary state to any directional state were uniformly distributed and were not affected by the presence of bFGF. PMID- 8703418 TI - Response of single, pairs, and clusters of epithelial cells to substratum topography. AB - Cells cultured on grooved substrata change their shape, orientation, and direction of locomotion in response to substratum topography, a phenomenon called contact or topographic guidance. Porcine epithelial cells (E-cells) spread on micromachined grooved or smooth control surfaces were examined by epifluorescence and confocal microscopy to determine area, cell shape, and orientation in conjunction with distributions and orientations of actin filaments and microtubules. Single cells, cells within a pair or cluster, and pairs or clusters considered as a unit were compared. As expected, cell contact increased cell spreading, but surprisingly, increased cell contact influenced cell shape on smooth and grooved surfaces and increased alignment of cells spread on grooves. Both actin filaments and microtubules aligned initially and most consistently along the walls and ridge-groove edges. Single E-cells displayed the least variability of aligned cytoskeletal patterns. E-cells within clusters displayed the most variability as local topographic effects on the cytoskeleton could be overridden by adjacent cell contact. Overall, contact guidance of E-cells was neither synonymous with nor contingent upon an elliptical morphology oriented to the topography. E-cells also differed from fibroblasts in their response to cell contact and in their lack of a relationship between cell polarity and locomotion. PMID- 8703419 TI - Properties of circulating leukocytes in spontaneously hypertensive rats. AB - The factors responsible for predisposition to progressive organ injury and vascular complications in arterial hypertension are uncertain. Recent evidence shows that leukocytes participate in cardiovascular conditions for which hypertension is a risk factor. Therefore, there is a need to define the properties of circulating leukocytes in hypertensives. There are about twice as many circulating leukocytes in spontaneous hypertensive rats (SHRs) compared with their normotensive controls, the Wistar-Kyoto rats (WKYs). The SHR neutrophils are viscoelastic and similar to neutrophils in WKYs but exhibit lower deformability in short-term elastic deformation. Mature SHRs have elevated levels of spontaneous pseudopod formation. Mild stimulation with N-formyl-Met-Leu-Phe or platelet-activating factor (10(-8) M) results in a significantly enhanced level of neutrophil pseudopod formation in SHRs but not in WKYs. SHRs exhibit higher levels of spontaneous superoxide formation. Alkaline phosphatase content of individual circulating neutrophils in SHRs is on average lower while plasma levels of alkaline phosphatase in the same samples are elevated in the SHRs. Spontaneous degranulation of SHR neutrophils is also detectable with myeloperoxidase measurements. Such activity of circulating leukocytes poses a significant risk for vascular cytotoxicity in the hypertensive rats. PMID- 8703420 TI - Changes in surface topography in endothelial monolayers with time at confluence: influence on subcellular shear stress distribution due to flow. AB - It is well known that the morphology of endothelial cells in culture changes dramatically as they go from subconfluence to confluence. After reaching confluence, however, the morphology continues to change but much more subtly as cell density increases and they become more uniform in size and shape. Measurements of surface topography of confluent cells by atomic force microscopy (AFM) showed that cell heights became more uniform and that the root-mean-square amplitude of surface undulations decreased by 7% compared with monolayers that had just reached confluence. Computational fluid dynamics simulations of flow over the endothelial surface geometries measured by AFM showed that the change in topography with time after confluence altered the shear stress distribution, resulting in an increase in the stress concentrations experienced by the cells despite the reduced amplitude of the surface undulation. These data suggest that the starting point for in vitro experiments may influence the measured responses to shear stress, particularly transient responses that occur before structural adaptation takes place. In addition, changes in surface topography may reflect changes in cell tension, cytoskeletal structure, and adhesion to the substratum, all of which are associated with the regulation of growth in anchorage-dependent cells. PMID- 8703421 TI - Insulin does not disrupt actin microfilaments, microtubules, and in vitro aortic endothelial wound repair. AB - In the face of small denuding injuries, the endothelium undergoes a process of rapid repair involving actin microfilaments, microtubules, and centrosomes to reestablish an intact monolayer. Failure to maintain an intact endothelial monolayer is an important factor in the pathogenesis of the atherosclerotic plaque. It was hypothesized that increased susceptibility to atherosclerosis in diabetes mellitus may be, in part, due to delayed reendothelialization following endothelial injury. To test this, the effects of high insulin concentrations on the reendothelialization of small wounds were examined using an in vitro porcine aortic endothelial cell wound model. Elevated concentrations of insulin did not disrupt the confluent endothelial monolayer or alter endothelial cell shape. Insulin also did not induce detectable alterations in the distribution of microtubules and microfilaments in the confluent monolayer. High insulin did not reduce the extent of reendothelialization of a linear wound made in the confluent monolayer. Centrosomal reorientation was similar to that of control wounded cultures as was the reorganization of the microfilaments and microtubules. The data suggest that the atherogenic effects of hyperinsulinemia are not due to disruption of endothelial repair. PMID- 8703422 TI - Endotoxin, tumor necrosis factor, and dexamethasone effects on human endothelial cell fibronectin dynamics: synthesis, matrix assembly, and receptor expression. AB - The three inflammatory modulators endotoxin, tumor necrosis factor (TNF) alpha, and dexamethasone (DEX) were studied for their effects on fibronectin (FN) dynamics in human umbilical vein endothelial cells. Cell culture supernatants were analyzed for new soluble pool FN synthesis. Endotoxin (LPS) (10 micrograms/mL) decreased the newly synthesized soluble pool of FN (p < 0.05). An increase in soluble FN was demonstrated with 1 and 10 ng/mL TNF alpha (p < 0.05). DEX decreased newly synthesized endothelial cell (EC) FN in the soluble pool at 4, 40, and 400 micrograms/mL (p < 0.05). Extracellular matrix FN content was examined using immunofluorescence. The thick FN mesh seen in control cells contrasted with a decreased FN matrix after treatment with each of the three study agents. Immunoprecipitation of the FN receptor alpha 5 beta 1 integrin from [35S]methionine-labelled cell extracts demonstrated down regulation of receptor expression by both TNF alpha and DEX as compared with control samples. These data indicate that LPS, TNF alpha, and DEX may weaken EC-substratum adhesion by differential effects on FN synthesis and secretion, FN incorporation into the extracellular matrix, and down regulation of FN receptor expression. PMID- 8703423 TI - Mechano-reception in osteoblast-like cells. AB - Response to mechanical stimulation is a basic biological phenomenon. Nearly all cells process mechanical input and respond to it by inducing and modulating biochemical pathways. In organisms with tissues, if the average mechanical load is increased, some tissues can increase their performance and often increase their bulk by cell division. A reduction in mechanical loading decreases performance, catabolic activity gains, and the tissue degenerates. The process of anabolism and catabolism regulated by mechanical loading is a second-to-second, minute-to-minute, and hour-to-hour process that works together with local and systemic hormones to ensure that the tissue can meet the demands of the mechanical environment. On the other hand, a mechanical load that is too high can cause tissue and matrix failure and damage to the cells, which can result in inflammation. In this paper, we review the possible biophysical and cell biological mechanisms that might be responsible for transducing physiological and hyperphysiological mechanical loading into the biological response of skeletal cells. We speculate on what the mechanism of mechano-transduction in bone might be compared with that of other cells and on how information produced by mechanical loading might be passed on to other cells to achieve a coordinated tissue response. PMID- 8703424 TI - Regulation of cell volume and intracellular pH in hyposmotically swollen rat osteosarcoma cells. AB - The maintenance of cell volume involves transduction of a volume-sensing signal into effectors of volume-regulatory transporters. After exposure to anisotonic conditions, cells undergo compensatory volume changes that are mediated by active transport and passive movement of ions and solutes. Intracellular pH (pHi) homeostasis may be compromised during these processes. We have studied pHi and some of the signal transduction mechanisms involved in the regulatory volume decrease (RVD) that occurs after exposure to hypoosmolar conditions in rat osteosarcoma cells, ROS 17/2.8. Cells were loaded with BCECF; pHi and cell volume were estimated by dual excitation ratio fluorimetry. Swelling of cells in 4-(2 hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) buffered hypotonic medium induced a rapid cell swelling followed by an incomplete RVD of approximately 30% in suspended (i.e., round) cells and approximately 60% in attached (i.e., spread) cells that was independent of subpassage number. RVD was inhibited by ouabain, valinomycin, and high external [K+], all of which should reduce the cell membrane electrochemical gradient for K+. Inhibition of RVD was induced also by decreasing intracellular [Ca2+] with BAPTA-AM and by depletion of Cl-, indicating the role of calcium-regulated K+ and Cl- efflux during RVD. Depolymerization of actin filaments by cytochalasin D prolonged the RVD three-fold and nonspecific activation of GTP-binding proteins up-regulated RVD. In attached cells the hypoosmolar-induced swelling caused a large reduction in pHi (approximately 0.7 units), which was sustained as long as cells were in hypoosmotic medium. The reduction of pHi induced by cell swelling was inhibited by Na(+)-free extracellular medium, ouabain, the tyrosine kinase inhibitor genistein, and to a lesser extent by Cl(-)-free medium. However, amiloride failed to inhibit the hypoosmolar-induced reduction of pHi. Collectively these data indicate that RVD of ROS 17/2.8 cells in HEPES-buffered medium is dependent on conductive efflux of K+ and Cl- that is regulated by cell shape, actin, and GTP-binding proteins. The sustained inhibition of pHi homeostasis induced by cell swelling may reflect the existence of cell volume sensing mechanisms that operate through tyrosine kinases to regulate pHi. PMID- 8703425 TI - Cytoskeletal mechanics of neurulation: insights obtained from computer simulations. AB - The morphogenetic movements associated with the process of neurulation have been the subject of much investigation during the last one hundred years. A plethora of experimental evidence has been generated regarding the forces that drive this seemingly simple process, and many theories about the mechanics of the process have been proposed. Recent computer simulations have proved useful for evaluating these theories from a mechanical perspective. In this work, computer simulations are used to investigate several theories about the forces that drive neurulation. A simplified version of a formulation previously presented by the authors provides the mathematical foundation for these simulations. The simulations confirm that forces generated by circumferential microfilament bundles (CMB's) in conjunction with notochord forces can produce the rolling motions characteristic of amphibian neurulation. They also support the notion that redundancies exist in the systems of forces available to drive neurulation shape changes. The shape changes that occur following a variety of surgical and teratogenic interventions are also simulated. These simulations corroborate the role of circumferential microfilament bundles as a primary force generator. PMID- 8703426 TI - Biomechanical feedback in morphogenesis, as exemplified by stretch responses of amphibian embryonic tissues. AB - We explore the idea that morphogenetical processes may be self-regulated by the biomechanical feedback established between the active stress-generating devices and the passive stresses of stretching and (or) compression, these feedback directed towards hyperrestoration (restoration with overlapping) of the initial stress values. As an example, a stretch-induced behaviour of the pieces of ventral ectoderm of Xenopus laevis early gastrulae is considered. By stretching the explants in 1.3-1.7 times, we induced several active poststretching cell responses, including further autonomous elongation of an explant in the stretch direction and contraction in the perpendicular direction, as well as more complicated shape changes. At the cellular level, these responses were associated with the return of the stretched cells to isodiametrical shapes and with the production of extensive cell protrusions along the stretch direction. As shown by dissections, the stretch-induced tissue tensions were considerably diminished in the poststretching period. The results obtained are discussed within the framework of the hyperrestoration hypothesis. PMID- 8703427 TI - Morphogenesis and mechanical instability of a prestressed tissue. AB - We consider the issue of whether purely mechanical properties of biological systems can, in principle, play a significant role in morphogenesis. As a simple example, we model a spherically arranged epithelium that is symmetrically prestressed under the action of cytoskeletal elements. A three-dimensional exact bifurcation analysis indicates the existence of a critical radius beyond which, for a physiologically attainable prestress, the spherical organoid is mechanically unstable and will buckle. We conclude that the purely mechanical aspects of biological tissues may indeed play a role in morphogenesis. PMID- 8703428 TI - Development of a defined medium fermentation process for physostigmine production by Streptomyces griseofuscus. AB - Physostigmine is a plant alkaloid of great interest as a therapeutic candidate for the treatment of Alzheimer's disease. Fortunately, this compound is also produced by Streptomyces griseofuscus NRRL 5324 during submerged cultivation. A fermentation process that used chemically defined medium was therefore developed for its production. By means of statistical experimentation, the physostigmine titer was quickly increased from 20 mg/l to 520 mg/l with a culture growth of 19 gl dry cell weight on the shake-flask scale. Further medium optimization resulted in a yield of 790 mg/l in a 23-1 bioreactor using a batch process. A titer of 880 mg/l was attained during scale-up in a 800-1 fermentor by employing a nutrient feeding strategy. This production represents a 44-fold increase over the yield from the initial process in shake-flasks. The defined-medium fermentation broth was very amenable to downstream processing. PMID- 8703429 TI - Molecular analysis of the gene cluster involved in cephalosporin biosynthesis from Lysobacter lactamgenus YK90. AB - Determination of the nucleotide sequence downstream from the Lysobacter lactamgenus pcbC gene encoding isopenicillin N synthase revealed that five open reading frames (ORF) including the pcbC gene were tightly linked in the same orientation. Each ORF has the remarkable feature of the protein-coding frame in the DNA sequence with a high G+C content. Expression in Escherichia coli and a comparison of the deduced amino acid sequences with published sequences showed that the gene cluster contained a deacetoxycephalosporin C synthetase (DAOCS) gene (cefE), an ORF having homology with the Cephalosporium acremonium DAOCS/deacetylcephalosporin C synthetase gene (cefEF), an isopenicillin N epimerase gene(cefD), and a beta-lactamase gene. The gene order was pcbC-cefE ORF3-cefD-beta-lactamase. PMID- 8703430 TI - Mutants blocked in penicillin biosynthesis show a deletion of the entire penicillin gene cluster at a specific site within a conserved hexanucleotide sequence. AB - The organization of the genes of the penicillin cluster has been studied in three different mutants of P. chrysogenum impaired in penicillin biosynthesis. The three blocked mutants (derived from the parental strain P. chrysogenum Bb-1) lacked the genes pcbAB, pcbC and penDE of the penicillin biosynthetic pathway and were unable to form isopenicillin N synthase and isopenicillin N acyltransferase. All strains were identified as P. chrysogenum derivatives by fingerprinting analysis with (GTG)n as a probe. The borders of the deleted region were cloned and sequenced, showing the same junction point in the three mutants. The deleted DNA region was found to be identical to that described in P. chrysogenum npe10. The frequent deletion of the pen gene cluster at this point may indicate that this cluster is located in an unstable genetic region, flanked by hot spots of recombination, that is easily lost by mutagen-induced recombination. PMID- 8703431 TI - Characterization of the cefF gene of Nocardia lactamdurans encoding a 3' methylcephem hydroxylase different from the 7-cephem hydroxylase. AB - The cefF gene of Nocardia lactamdurans, encoding a functional 2-oxoglutarate dependent 3'-methylcephem hydroxylase (deacetoxycephalosporin C hydroxylase) has been found to be closely linked to the pcbC gene in the cephamycin C gene cluster. The open-reading frame is 933 bp long and could encode a protein of M(r) 34,366. Introduction of cefF in the cephamycin-non-producer Streptomyces lividans conferred 3'-methylcephem-hydroxylating activity to the transformants but did not result in hydroxylation at carbon 7 of cephamycin. No 3'-methylcephem hydroxylase activity was observed when the cefF gene was introduced in S. lividans in pIJ699 (a vector containing two transcriptional terminators that prevent read-through expression), which suggests that this gene lacks an independent promoter. PMID- 8703432 TI - Application of Ty1 for cloned gene insertion: amplification of a large regulated expression cassette in Saccharomyces cerevisiae. AB - A large cassette, 4.6 x 10(3) bases (4.6 kb) in length, containing an inducible expression system (the yeast CUP1 promoter fused to the Escherichia coli lacZ structural gene) and a bacterial neomycin-resistance gene (neo) has been cloned into the noncoding region of a GAL1-regulated Ty1 retrotransposon. Galactose was used to induce retrotransposition in Saccharomyces cerevisiae, and cells containing integrations were selected by resistance to the aminoglycoside G418. Integrations of neo and CUP1p-lacZ were verified, and beta-galactosidase activity was confirmed. Analysis via Southern blots demonstrated integrations at various chromosomal locations, and the number of insertions obtained ranged from one to five after three rounds of induction. Therefore, the packaging limit of Ty1 virus like particles for RNA is at least 10.3 kb and Ty1 can transpose foreign genes as large as 4.6 kb, demonstrating the practical application of Ty1 for the insertion of large regulated expression cassettes. PMID- 8703433 TI - Recombinant outer-surface protein A (des-Cys1-OspA) from the Lyme disease spirochete Borrelia burgdorferi: high production levels in Saccharomyces cerevisiae yeast cultures. AB - The recombinant outer-surface protein A with an N-terminally truncated form (des Cys1-OspA) from the Lyme disease spirochete Borrelia burgdorferi was expressed in Saccharomyces cerevisiae at high production levels. Since the recombinant vaccine candidate expressed in Escherichia coli exhibits low production yields and the purification of lipoproteins appears to be difficult, we have investigated the secretion of a soluble recombinant OspA in the yeast S. cerevisiae. In this way, a Leu+ derivative of S. cerevisiae cI3ABYS86 was used as the host strain transformed with an expression plasmid containing the gene encoding des-Cys1-OspA and driven by the MF alpha 1 promoter. The fed-batch culture results revealed that an efficient secretion of des-Cys1-OspA is obtained with a high production level of about 2.1 g l-1 at a cell density of 101 g l-1 cell dry weight. The accumulation of recombinant protein in the supernatant exceeds 6% of the total yeast proteins when estimated by sodium dodecyl sulphate/polyacrylamide gel electrophoresis. Moreover, des-Cys1-OspA showed lower solubilities at high cell densities and, as a consequence, a fraction of the recombinant protein precipitated. An internal cleavage of the MF alpha 1 pro::des-Cys1-OspA precursor was also detected. However, in this case the cleavage occurred at a frequency such that the large amounts of the secreted des-Cys1-OspA could be employed for the evaluation of an immunogenic effect on animal immunization. These studies will extend the knowledge of the usefulness of OspA as a vaccine for Lyme borreliosis. PMID- 8703434 TI - Influence of phytogenic surfactants (quillaya saponin and soya lecithin) on bio elimination of phenanthrene and fluoranthene by three bacteria. AB - The influence of two phytogenic surfactants on the elimination of polycyclic aromatic hydrocarbons (PAH) was studied in shaken-batch cultures of three soil bacteria under axenic conditions. At sufficiently high concentrations, quillaya saponin and soybean lecithin solubilized phenanthrene or fluoranthene efficiently. However, complete solubilization of the PAH by lecithin only doubled the maximal rate of elimination of the two PAH compounds by Pseudomonas 0259, strain MKm (Rhizomonas ?) and Mycobacterium EMI 2. By contrast, quillaya saponin did not improve PAH bioavailability, and in strain MKm it caused significant growth lags above 2.5 g/l. Simultaneously with the elimination of the PAH the bacteria utilized the surfactants as substrates for growth. Intermediate formation of PAH metabolites was noted. The results suggest that some phytogenic surfactants might improve PAH bioavailability in rhizospheres. PMID- 8703435 TI - Microbial degradation of polycyclic aromatic hydrocarbons in soils affected by the organic matrix of compost. AB - This paper describes the degradation of naphthalene, phenanthrene, anthracene, fluoranthene, and pyrene in soil and soil/compost mixtures. Compost addition facilitated the degradation of 500 mg naphthalene/kg soil and 100 mg/kg each of other polycyclic aromatic hydrocarbons (PAH) within 25 days in soil systems with water contents below the water-holding capacity. By means of a humic acid extraction, it was demonstrated that the decrease of PAH concentrations after compost addition was not caused by a sorption to organic matter preventing PAH analysis. The enhanced PAH degradation was examined in a series of batch experiments with contaminated soil to evaluate whether the effect of compost addition is caused by the microorganisms of the compost itself, by the properties of the organic matrix of the compost material, or by water-soluble fertilising substances. The experiments revealed that the release of fertilising substances from the compost and the shift of soil pH brought about by the compost did not cause the stimulatory effect. The microorganisms inherent to the compost were also not necessary for the enhanced degradation. Sterilised compost was recolonised by soil microorganisms after a lagphase yielding a degradation activity similar to that of the non-sterilised compost. The presence of the solid organic matrix of the compost seemed to be essential for the enhanced degradation. The soil/compost microflora, which was separated from the organic matrix in liquid cultures, exhibited a much lower degrading activity than in the presence of the solid organic material. PMID- 8703436 TI - Modelling for waste water treatment by Rhodopseudomonas palustris Y6 immobilized on fibre in a columnar bioreactor. AB - A kinetic model of continuous treatment of waste water by Rhodopseudomonas palustris Y6 immobilized on soft fibre in a columnar bioreaction system was established. Good agreement was found between the model prediction and the experimental data from continuous operation [initial chemical oxygen demand (COD) concentration = 29.700 g/l] of the system. The optimum operational conditions for the maximum COD reduction capacity were investigated from the model prediction and the experimental data. The waste water treatment process may significantly increase the waste reduction capacity because a large amount of active biomass for COD reduction is immobilized in the system, resulting in operation stability. The results presented here provide a useful basis for further scaling up and efficient operation of waste water treatment processes. PMID- 8703437 TI - Isolation of hexavalent chromium-reducing anaerobes from hexavalent-chromium contaminated and noncontaminated environments. AB - Hexavalent chromium [Cr(VI)], is a toxic, water-soluble contaminant present in many soils and industrial effluents. Bacteria from various soils were examined for Cr(VI) resistance and reducing potential. Microbes selected from both Cr(VI) contaminated and -noncontaminated soils and sediments were capable of catalyzing the reduction of Cr(VI) to Cr(III) a less toxic, less water-soluble form of Cr, demonstrating the utility of using a selection strategy for indigenous Cr(VI) reducing bacteria in a bioprocess. As a result, indigenous Cr(VI)- reducing microbes from contaminated sites should provide the means for developing a bioprocess to reduce Cr(VI) to Cr(III) in nonsterile effluents such as those from soil washes. This approach also avoids the contamination problems associated with pure cultures of allochthonous microorganisms. In addition the apparent ubiquity of Cr(VI)-reducing bacteria in soil and sediments indicates potential for in situ bioremediation of Cr(VI)-contaminated soils and ground water. PMID- 8703438 TI - Biological significance of IgA1 proteases in bacterial colonization and pathogenesis: critical evaluation of experimental evidence. AB - IgA1 protease activity, which allows bacteria to cleave human IgA1 in the hinge region, represents a striking example of convergent evolution of a specific property in bacteria. Although it has been known since 1979 that IgA1 protease is produced by the three leading causes of bacterial meningitis in addition to important urogenital pathogens and some members of the oropharyngeal flora, the exact role of this enzyme in bacterial pathogenesis is still incompletely understood owing to lack of a satisfactory animal model. Cleavage of IgA1 by these post-proline endopeptidases efficiently separates the monomeric antigen binding fragments from the secondary effector functions of the IgA1 antibody molecule. Several in vivo and in vitro observations indicate that the enzymes are important for the ability of bacteria to colonize mucosal membranes in the presence of S-IgA antibodies. Furthermore, the extensive cleavage of IgA sometimes observed in vivo, suggests that IgA1 protease activity results in a local functional IgA deficiency that may facilitate colonization of other microorganisms and the penetration of potential allergens. It has been hypothesized that IgA1 protease activity of Haemophilus influenzae, Neisseria meningitidis, and Streptococcus pneumoniae, under special immunological circumstances, allows these bacteria to take advantage of specific IgA1 antibodies in a strategy to evade other immune factors of the human body. The decisive factor is the balance between IgA antibodies against surface antigens of the respective bacteria and their IgA1 protease. Recent studies have shown that serine-type IgA1 proteases of H. influenzae, meningococci, and gonococci belong to a family of proteins used by a diverse group of Gram-negative bacteria for colonization and invasion. PMID- 8703439 TI - The effect of oral commensal bacteria on candidal adhesion to denture acrylic surfaces. An in vitro study. AB - The effect of four different species of oral bacteria (Streptococci (X 2); Escherichia; Porphyromonas) on the adhesion of Candida albicans and Candida krusei to denture acrylic surfaces was studied using a previously described in vitro adhesion assay. Clear acrylic strips immersed in known concentrations of bacterial suspensions and incubated for 45 min to 1 h, at 37 degrees C, were transferred to yeast suspensions of known concentrations and incubated for a similar period, and the adherent yeasts were quantified using an image analysis system. Pre-exposure to different bacterial concentrations resulted in suppression of candidal adhesion to varying degrees. Of three selected concentrations, exposure to the highest bacterial concentration (10(6) organisms/ml) resulted in a consistent reduction in candidal adhesion, except for E. coli-C. albicans and S. salivarius-C. albicans combinations. Candidal adhesion was significantly reduced on pre-exposure to S. sanguis and P. gingivalis, but not E. coli. There were also significant differences in relative adhesion between C. albicans and C. krusei. Taken together, these results indicate that the adhesion of yeasts to acrylic strips is modulated both by the quantity and the quality of pre-existing bacterial flora on acrylic surfaces. PMID- 8703440 TI - Effects of Chinese medicinal herbs on a rat model of chronic Pseudomonas aeruginosa lung infection. AB - The aim of the study was to evaluate the effects of two kinds of Chinese medicinal herbs, Isatis tinctoria L (ITL) and Daphne giraldii Nitsche (DGN), on a rat model of chronic Pseudomonas aeruginosa lung infection mimicking cystic fibrosis (CF). Compared to the control group, both drugs were able to reduce the incidence of lung abscess (p < 0.05) and to decrease the severity of the macroscopic pathology in lungs (p < 0.05). In the great majority of the rats, the herbs altered the inflammatory response in the lungs from an acute type inflammation, dominated by polymorphonuclear leukocytes (PMN), to a chronic type inflammation, dominated by mononuclear leukocytes (MN). DGN also improved the clearance of P. aeruginosa from the lungs (p < 0.03) compared with the control group. There were no significant differences between the control group and the two herbal groups with regard to serum IgG and IgA anti-P. aeruginosa sonicate antibodies. However, the IgM concentration in the ITL group was significantly lower than in the control group (p < 0.03). These results suggest that the two medicinal herbs might be helpful to CF patients with chronic P. aeruginosa lung infection, DGN being the most favorable. PMID- 8703441 TI - Number of activated T-helper cells and NK cells in peripheral blood is decreased in severe Crohn's disease. AB - Monoclonal antibodies and flow cytometry were used to analyse the proportions of T-cell subsets and NK cells in blood of patients (n = 45) with Crohn's disease. In patients with severe activity disease decreased numbers of activated (CD25+CD4+) T-helper cells and NK (CD16+CD56+) cells were found, while in patients with low activity disease the numbers of activated T-helper cells were increased and the numbers of NK cells were similar to those in normal controls. Thus, 8% of T cells were CD25+CD4+ and 16% of mononuclear cells were CD16+CD56+ in patients with severe disease. In patients with quiescent disease, 11% of T cells were CD25+CD4+ and 26% of mononuclear cells were CD16+CD56+. The results suggest that disease activity may be reflected in the proportions of blood circulating mononuclear cells, perhaps because of accumulation of CD25+CD4+ T helper cells and NK cells in the affected tissue during exacerbation of the disease. PMID- 8703442 TI - Enterochromaffin cell density in the gastric mucosa of patients with chronic renal failure. AB - Thirty patients with chronic renal failure (CRF) and 30 age- and sex-matched controls were assessed for gastrointestinal diseases by gastroscopy, serum gastrin determination, and routine clinical and laboratory evaluation. Biopsy specimens from their gastric oxyntic mucosa were immunohistochemically stained with monoclonal antibodies against serotonin (5-hydroxytryptamine) and chromogranin A, the latter staining all gastric endocrine cells, the former disclosing serotonin-containing enterochromaffin (EC) cells only. The average EC cell density (cells/mm2) in the CRF patients was significantly lower than in the controls: 2.6 vs 12.9 (p = 0.0005). The EC cell counts also correlated negatively with serum gastrin values (p = 0.0031). The densities of the chromogranin positive cells did not differ between CRF patients (74 cells/mm2) and controls (76 cells/mm2) (p = 0.7559). We conclude that, in addition to the previously known findings of hypoacidity, persistent hypergastrinaemia, and G and parietal cell hyperplasia, CRF also reduces the number of oxyntic EC cells. The negative correlation between EC cell density and serum gastrin levels reflects the complex interplay between different endocrinological activities in the gastrointestinal tract. PMID- 8703443 TI - The interference of gingival cell cultures with growth of selected bacteria. AB - The aim of the present study was to analyze the interference of oral tissue cells or cell lines (effector cells) with growth of reference bacteria, and furthermore to investigate whether cells derived from different individuals differ in such activity. The reference bacteria were Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus mitis, Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Fusobacterium nucleatum. The effector cells used were gingival fibroblasts (GF) from 21 periodontally involved persons, gingival epithelial cells (E) from 2 such persons, HeLa cells (HeLa), and an amnion cell line (Amnion). The cells were cultivated and their supernatants tested for antibacterial activity in a Bioscreen robot analyzer (Labsystems, Finland). Results suggest that the antibacterial activity of each tested primary cell line of tissue had its own profile depending on cell type and donor, and that the composition of oral microbiota was influenced by oral cells, which might, in turn, contribute to the variations in the pathogenesis of periodontal diseases. PMID- 8703444 TI - Ultrastructural changes in primary guinea pig osteoarthritis with special reference to collagen. AB - Collagen fibril distribution and surface and volume densities in proximal tibial articular cartilage were measured in 6- and 12-month-old Dunkin-Hartley guinea pigs developing primary osteoarthritis. At 12 months, gross fibrillation and ulceration of the articular cartilage were observed on the medial but not on the lateral condyle. Collagen volume density decreased with age in the interterritorial compartments in the superficial zone, medially by 16% and laterally by 8%. In the upper radial zone, collagen volume density decreased interterritorially by 10% on the medial condyle only. Despite gross osteoarthritic changes, only moderate and predominantly focal ultrastructural collagen changes were observed. Thus neither gross network disruption nor fibril thickening seems to be a general feature in early guinea pig osteoarthritis. PMID- 8703445 TI - Loss of protective immunity to polio, diphtheria and Haemophilus influenzae type b after allogeneic bone marrow transplantation. AB - Immunity to poliovirus, diphtheria and Haemophilus influenzae type b (Hib) was studied in 16 adult recipients of a bone marrow transplant from an HLA-identical sibling donor in order to evaluate the need for revaccinations. T-cell depletion was not done in any case. The donors and patients were studied before bone marrow transplantation (BMT) and the patients 1, 3, 6, and 12 months later. Prior to the BMT 10 of 11 patients were immune (titre > or = 4) to all vaccine poliovirus types by a standard microneutralization assay. At 12 months after BMT only two of seven patients were immune to all vaccine types, and none had immunity against an antigenically altered poliovirus type 3 strain Finland. The geometric means of antibody titres against poliovirus types 1, 2, and 3 strain Saukett and strain Finland declined gradually after 1 month postgrafting, being 4.4, 5.4, 3.3, and 1.3 respectively at 12 months after BMT. At 1 year 6 of 11 patients had immunity against diphtheria by a toxin neutralization method, but the antitoxin geometric mean level had decreased to a barely protective level, 0.01 IU/ml. The geometric mean Hib antibody concentration decreased during the first 6 months after BMT and thereafter increased slightly. A significant proportion of BMT recipients lose their protection against polio, diphtheria and Hib, and revaccinations are necessary. PMID- 8703446 TI - Imprint cytology of cat scratch disease. A report of eight cases. AB - The cytologic features of cat scratch disease (CSD) from eight cases in imprint smears are presented. All patients were clinicopathologically diagnosed with CSD as follows: 1) a history of animal exposure was recorded 2 to 4 weeks before lymphadenopathy; 2) the disease occurred in the autumn and winter months; 3) a characteristic histopathology in the biopsied lymph node specimens was observed; and 4) Warthin-Starry silver stain-positive bacteria were detected in four of the seven cases examined. The characteristic cytologic finding was the presence of confluent epithelioid cells with nearby and central scattering of neutrophils against a background of polymorphic inflammatory cells. Furthermore, a varying number of medium-sized to large lymphoid cells with an appearance suggestive of monocytoid B lymphocytes (MBLs) were noted to be associated with the epithelioid cells. These cytologic findings closely paralleled the histologic patterns of epithelioid cell granulomas, with and without MBLs, which we have previously reported are probably associated with the disease. PMID- 8703447 TI - Intimal thickening and fragmentation of the internal elastic lamina in the mesenteric arteries. AB - To evaluate the intimal thickness and continuity of the internal elastic lamina (IEL) in the mesenteric arteries, proximal segments of the coeliac artery (CA), superior mesenteric artery (SMA) and inferior mesenteric artery (IMA) were studied light microscopically in 123 consecutive postmortem examinations. The mean age of the subjects was 62 years. Arterial segments were fixed in formalin, embedded in paraffin blocks, sectioned, and stained with Masson's trichrome. Fifty-one per cent of the samples examined showed at least minimal intimal thickening. The occurrence of significant luminal narrowing increased with age. We found a positive correlation between defects in the IEL and thickness of the intima in the mesenteric arteries, which is in harmony with previous observations showing marked fragmentation of the IEL in atherogenesis. PMID- 8703448 TI - The ROMI protocol: do we have a rational approach? PMID- 8703449 TI - Cryoprostatectomy consistently elevates serum creatine kinase-MB isoenzyme. AB - STUDY OBJECTIVE: To measure serum CK-MB, a market of myocardial infarction (MI), in elderly men before and after cryoprostatectomy. DESIGN: Serum CK-MB was measured on each patient before and after cryoprostatectomy. Each patient's preoperative result was used as control measurement for comparison with measurements made after cryoprostatectomy. SETTING: Inpatient operating room and postanesthetic recovery unit of a university-affiliated general hospital. PATIENTS: 38 male patients, mean (SEM) age 69.1 +/- 1.4 years, undergoing cryoprostatectomy. INTERVENTIONS: All patients had a 12-lead ECG prior to surgery, in the recovery room, and 24 hours after surgery. Serum CK-MB was measured prior to induction of anesthesia, on arrival in the recovery room, and at 8 and 24 hours after surgery. Lactate dehydrogenase (LDH) and isoenzymes also were measured in 10 patients before and after cryoprostatectomy. MEASUREMENTS AND MAIN RESULTS: All patients underwent uneventful cryoprostatectomy. No patients had new ECG changes after surgery. All patients had normal serum CK and CK-MB concentrations before surgery. Serum CK and CK-MB were significantly elevated after cryoprostatectomy (p < 0.001). Enzyme values were greatest 8 hours after surgery: total CK mean 1453 +/- 145 U/L (range 199 to 3,356 U/L); CK-MB mean 52 +/- 3 ng/ml (range 12 to 114 ng/ml) or 5.0 +/- 0.5% of total CK (range 1.6% to 12.4%). All patients had significant elevations of LDH after cryoprostatectomy but did not show an increase in the ratio of LDH1 to LDH2 isoenzymes. Finally, unlike patients with an acute MI, the activity of CK-MB isoenzyme when measured by gel electrophoresis was two to three times greater (mean, 2.6 +/- 0.7) than the concentration measured with the monoclonal antibody assay in patients after cryoprostatectomy. CONCLUSION: Serum CK-MB is an unreliable test to diagnose an MI in patients who have undergone cryoprostatectomy. PMID- 8703450 TI - Intravenous trimethaphan during epidural plus general anesthesia decreases the direct radial artery pressure lower than the brachial artery pressure. AB - STUDY OBJECTIVE: To determine whether vasodilators such as sodium nitroprusside (SNP) and trimethaphan (TMP) produce a pressure difference between the radial artery and the brachial artery during epidural plus general anesthesia or simple general anesthesia. DESIGN: Randomized study and prospective study. SETTING: Operating rooms of two hospitals. PATIENTS: 36 and 6 adult patients in the first and second studies, respectively, who received spherical acetabular osteotomy with induced hypotensive anesthesia. INTERVENTIONS: In the first study, 18 patients received epidural plus general anesthesia (epidural group) and 18 patients received general anesthesia alone (general group). All patients received the hypotensive drugs for more than 50 minutes each. In the second study, 6 patients received TMP-induced hypotension for 20 minutes under epidural plus general anesthesia. MEASUREMENTS AND MAIN RESULTS: In the first study, radial intra-arterial blood pressure (RIBP) and tonometric blood pressure (TBP) calibrated with oscillometric blood pressure of the arm were compared. In the second study, RIBP and the brachial intra-arterial blood pressure (BIBP) were compared. In the first study, the bias between RIBP and TBP for systolic, mean and diastolic blood pressure were significantly less during TMP-induced hypotension in the epidural group (-11.5 +/- 2.5 (mean +/- SD), -6.0 +/- 3.1, and -2.8 +/- 3.7 mmHg, respectively] than during SNP-induced hypotension in the epidural group and SNP- and TMP-induced hypotension in the general group (p < 0.01). The precision of systolic and mean pressures were significantly greater during TMP-induced hypotension in the epidural group (11.8 +/- 2.3 and 7.1 +/- 1.9 mmHg, respectively) than the other three hypotension groups (p < 0.01). In the second study, the bias between RIBP and BIBP for systolic, mean, and diastolic pressures were significantly less (p < 0.01), and precision was significantly greater during hypotension than during normotension (p < 0.01). CONCLUSIONS: Our results demonstrate that TMP decreases the direct radial artery systolic and mean pressures to levels below the brachial artery systolic and mean pressures in patients who received epidural plus general anesthesia. PMID- 8703451 TI - A comparison of sevoflurane with halothane in outpatient adenotomy in children with mild upper respiratory tract infections. AB - STUDY OBJECTIVE: To investigate the efficacy and safety of sevoflurane compared with halothane in pediatric outpatient ear-nose-throat (ENT) surgery during the induction, maintenance, emergence, and recovery of anesthesia. DESIGN: Prospective, randomized, comparative, open-label study. SETTING: ENT operating room and postoperative recovery room at a university medical center. PATIENTS: 41 ASA status I and II children between the ages of 2 to 10 years, with mild upper respiratory tract infection (URI). INTERVENTIONS: Induction and maintenance of anesthesia with either sevoflurane or halothane for outpatient adenotomy, otomicroscopy, and myringotomy. MEASUREMENTS AND MAIN RESULTS: Induction (means +/- SEM) was significantly shorter in the sevoflurane group (2.6 +/- 0.2 minutes) than in the halothane group (3.2 +/- 0.2 minutes). There was no difference between the two groups with regard to complications that occurred during the induction and maintenance period. The time to emergence and recovery was significantly shorter with sevoflurane than with halothane (means +/- SEM; time to extubation 9.9 +/- 0.98 minutes vs. 13.4 +/- 1.06 minutes, time to eye opening 12.9 +/- 1.6 minutes vs. 24.5 +/- 1.8 minutes, command response time 20.7 +/- 2.5 minutes vs. 36.4 +/- 2.8 minutes). No difference in the incidence of complications during emergence and recovery was found. Evaluation of recovery as assessed by a modified Aldrete score showed that children who had received sevoflurane reached higher scores in the first 30 minutes following the discontinuation of the anesthetic. The Pain/Discomfort Scale demonstrated a difference in the sevoflurane group, with more children being agitated and restless. CONCLUSION: Sevoflurane provides a safe and rapid anesthetic induction with no differences in complications during the induction, maintenance, and emergence period. With sevoflurane, the time of emergence and recovery was significantly shorter. The characteristics of sevoflurane as evaluated in the present study make it a suitable anesthetic in pediatric outpatient surgery even in the presence of mild URI. PMID- 8703452 TI - Neurocirculatory responses to intubation with either an endotracheal tube or laryngeal mask airway in humans. AB - STUDY OBJECTIVE: To compare the sympathetic and hemodynamic responses to intubation with either an endotracheal tube (ETT) or laryngeal mask airway (LMA). DESIGN: Prospective, randomized, single-blinded study. SETTING: The in vivo study was carried out in an experimental laboratory. PATIENTS: 16 healthy male consenting volunteers, ages 20 to 31 years, were studied. INTERVENTIONS: After placement of a radial artery catheter, ECG electrodes, and a recording needle in the peroneal nerve, subjects were anesthetized with propofol 2.5 mg/kg, paralyzed with vecuronium 0.15 mg/kg, and ventilated via mask for 5 minutes with oxygen and 0.5 MAC desflurane or 0.5 MAC isoflurane. A LMA or ETT was inserted and neurocirculatory responses were continuously recorded. MEASUREMENTS AND MAIN RESULTS: Measurements of heart rate (HR), mean arterial pressure (MAP), and sympathetic nerve activity (SNA) were made at preintubation baseline and at the peak response after airway manipulation. The time to recovery to 20% and 10% of baseline MAP and HR also was measured. Neurocirculatory variables did not differ in either the LMA (n = 7) or ETT (n = 9) groups immediately prior to intubation. The ETT group demonstrated a 27% HR increase and a 42% MAP increase compared with a 12% HR increase and a 23% MAP increase in the LMA group. Muscle SNA increased 600% in the ETT group versus 66% in the LMA group (p < 0.01). The time to return MAP and HR to 20% and 10% of perintubation baseline was significantly longer in the ETT than the LMA group (p < 0.01). CONCLUSIONS: Because of the substantial reduction in the neurocirculatory responses to the LMA versus ETT, the LMA may prove advantageous in patients in whom HR and MAP increases may predispose to adverse cardiac or cerebrovascular events. PMID- 8703453 TI - The effect of laryngeal mask cuff pressure on postoperative sore throat incidence. AB - STUDY OBJECTIVE: To study the effect of laryngeal mask airway (LMA) cuff pressure on the incidence of postoperative sore throat. DESIGN: Prospective, randomized, observational study. SETTING: Operating room of a university hospital. PATIENTS: 200 consecutive adult patients requiring anesthesia for gynecologic procedures. INTERVENTIONS: Anesthesia was induced with thiopental 3-5 mg/kg, fentanyl 2 micrograms/kg, vecuronium bromide 0.05mg/kg, and enflurane 0.8% to 2% and maintained with nitrous oxide and oxygen (65%/35%) and enflurane. MEASUREMENTS AND MAIN RESULTS: In Group 1, cuff pressure measurement was continuously performed until the end of the operation. In Group 2, 5 minutes after induction of anesthesia and 2 minutes after insertion of the LMA, cuff pressure was also continuously observed and reduced to the minimal pressure required for airtightness. In the recovery room, after the operation, patients were questioned for postoperative sore throat 4, 8, and 24 hours after the operation following a scoring protocol (score 0 = no complaints, score 1 = minimal sore throat, score 2 = moderate sore throat, score 3 = severe sore throat: "never a LMA again". Continuous monitoring of cuff pressure revealed a steady increase of pressure (during the first 60 minutes increases of 43 cm H2O) in Group 1. In Group 2, after release of air, cuff pressures were significantly lower through the entire operation when compared with Group 1. In Group 1, 8 patients claimed to have a sore throat (Score 1, n = 4; Score 2, n = 3; Score 3, n = 1). In Group 2, no patient complained of sore throat. CONCLUSIONS: A significant increase in cuff pressure is seen during the first 60 minutes. Three minutes after insertion of the laryngeal mask, cuff pressure can significantly be reduced without any major gas leakage. Postoperative sore throat can be reduced when cuff pressure is continuously monitored and kept on low-pressure values. PMID- 8703454 TI - An effect of magnesium on neuromuscular function in parturients. AB - STUDY OBJECTIVE: To determine the effects of magnesium sulfate on neuromuscular transmission in parturients. DESIGN: Open-label, clinical study. SETTING: Preeclamptic patients at a university-affiliated hospital. PATIENTS: 12 pregnant patients, ages 18 to 40 years, with preeclampsia or entering preterm labor. INTERVENTIONS: Patients were treated with magnesium sulfate. Train-of-four (TOF) recordings of the adductor pollicis muscle were obtained before and 30 minutes after the magnesium sulfate infusion. MEASUREMENTS AND MAIN RESULTS: Each contractile response in the control TOF showed an increase in the tension developed in response to supramaximal stimulation. The TOF responses after magnesium sulfate did not demonstrate this phenomenon, but rather developed fade. The T4/T1 ratios ranged from 0.72 to 0.92. CONCLUSIONS: In this patient population, clinically relevant infusions of magnesium sulfate produced significant changes in neuromuscular transmission as manifested by loss of treppe phenomenon and diminished TOF response to ulnar nerve stimulation. PMID- 8703455 TI - Pulmonary vascular resistance and right ventricular function in morbid obesity in relation to gastric bypass surgery. AB - STUDY OBJECTIVE: To investigate right ventricular function (RVF) during gastric bypass surgery in morbidly obese patients. To study the influence of obstructive sleep apnea syndrome on hemodynamics and RVF as a preoperative evaluation in morbidly obese patients. DESIGN: Prospective study. SETTING: Surgical patients at a university hospital. PATIENTS: 14 morbidly obese patients undergoing gastric bypass surgery. INTERVENTIONS: Hemodynamic changes and RVF were investigated using a thermodilution ejection fraction volumetric catheter and the REF-1 computer. MEASUREMENTS AND MAIN RESULTS: Hemodynamic parameters and RVF were measured: (1) before induction of anesthesia, awake, (2) after induction of anesthesia, (3) after opening the abdomen, and (4) after closing the abdomen. Esophageal pressure was measured in the midesophagus after induction of anesthesia with a camino catheter and its device. Morbidly obese patients with obstructive sleep apnea syndrome had significantly lower PaO2, higher PaCO2, and higher pulmonary artery (PA) pressure and pulmonary vascular resistance (PVR) compared with patients without sleep apnea (p < 0.05). However, there were no significant differences in heart rate, mean blood pressure, pulmonary capillary wedge pressure (PCWP), right ventricular ejection fraction (RVEF), and right ventricular end-diastolic volume. During gastric bypass procedure, PA pressure, PCWP, and PVR decreased significantly after opening the abdomen (p < 0.05). There were no significant changes in RVEF and cardiac index during gastric bypass procedure. CONCLUSION: The presence of chronic hypoxemia and hypercarbia in our morbidly obese patients with obstructive sleep apnea syndrome while awake, causes significant increases in PA pressure and PVR. We also demonstrated that RVEF did not change significantly during gastric bypass procedure despite significant decreases in PA pressure, PCWP, and PVR after opening the abdomen. This decrease in PA pressure and PVR may be caused by decreases in pleural pressure reflected by a concomitant decrease in esophageal pressure. PMID- 8703456 TI - Sufentanil plasma concentrations following lower extremity tourniquet release. AB - STUDY OBJECTIVE: To investigate whether release of a tourniquet on the lower extremity affects plasma concentrations of sufentanil, as previously demonstrated with fentanyl and midazolam. DESIGN: Prospective. SETTING: University tertiary care institution with residency program. PATIENTS: 20 ASA status I, II, and III patients undergoing total knee arthroplasty under a tourniquet using a sufentanil, nitrous oxide, relaxant regimen. INTERVENTIONS: Each patient received sufentanil 1 to 2 micrograms/kg at induction of anesthesia and in 12.5 to 25 microgram increments as needed thereafter, until 15 minutes prior to tourniquet release. MEASUREMENTS AND MAIN RESULTS: Plasma sufentanil concentrations were determined before tourniquet inflation, immediately before tourniquet deflation, and 1, 2, 5, 10, 20, 30, and 40 minutes following deflation. A 15% elevation of plasma sufentanil concentration above that predicted by elimination pharmacokinetics defined a secondary peak. Although the aggregate data did not indicate an overall statistically significant rise in plasma concentrations after deflation, 9 (45%) patients exhibited a secondary peak in sufentanil plasma concentration following tourniquet deflation (range of secondary peaks, 16% to 89% above predicted values). No patient experienced clinically significant respiratory depression. CONCLUSION: Release of a tourniquet on the lower extremity may yield a detectable rise in plasma sufentanil concentration. PMID- 8703457 TI - Evaluation of pain following electrocautery tubal ligation and effect of intraoperative fentanyl. AB - STUDY OBJECTIVE: To evaluate pain following laparoscopic tubal sterilization (LTS) and the effects of supplemental intraoperative fentanyl. DESIGN: Open evaluation of pain intensity and opioid requirements with double-blind, randomized evaluation of intraoperative fentanyl. SETTING: Outpatient center of a university-affiliated teaching hospital. PATIENTS: 44 ASA status I patients who underwent LTS by electrocautery. INTERVENTIONS: Patients received either intravenous fentanyl (F) 1 microgram/kg or saline (S) at induction. MEASUREMENTS AND MAIN RESULTS: Pain intensity was measured with patient assessments of visual analog scales (VAS) and cumulative opioid requirements. The group that received intraoperative fentanyl received 70.2 +/- 3.7 micrograms at induction. VAS on postanesthesia care unit arrival was 4.9 +/- 0.5 (F) and 5.9 +/- 0.5 (S) (means +/- SEM). VAS after one hour was 2.1 +/- 0.2 in both groups. During that hour, the F group consumed 107.5 +/- 14.7 micrograms and the S group consumed 140.5 +/- 15.7 micrograms fentanyl. The range of fentanyl consumed by each group in the postoperative period was 0 to 250 micrograms. There were no statistically significant differences between the F and S groups. CONCLUSIONS: Pain following LTS by electrocautery is of significant magnitude and may require relatively large doses of opioids for adequate management. Supplemental intraoperative fentanyl 1 microgram/kg did not produce a clinically significant reduction in either pain scores or opioid requirements. PMID- 8703458 TI - Propofol permits tracheal intubation but does not affect postoperative myalgias. AB - STUDY OBJECTIVE: To determine the effect of propofol without succinylcholine on intubating conditions and postoperative myalgias in ambulatory surgical patients undergoing general anesthesia. DESIGN: Prospective, double-blind, randomized study. SETTING: Ambulatory surgery adult patients. PATIENTS: 56 ASA physical status I and II adult patients undergoing general endotracheal anesthesia. INTERVENTIONS: Group 1 patients received thiamylal plus succinylcholine, Group 2 patients received propofol plus succinylcholine; and Group 3 patients received propofol plus saline. All patients received fentanyl, lidocaine, and nitrous oxide plus isoflurane in oxygen. MEASUREMENTS AND MAIN RESULTS: Incidence and severity of fasciculations, tracheal intubating conditions, and myalgias on the first and third postoperative days were measured. Propofol did not affect the incidence or severity of fasciculations following succinylcholine, or the incidence of myalgias. Of patients who received propofol without succinylcholine, intubation was successful in 85%. CONCLUSIONS: Propofol did not affect the incidence or severity of postoperative myalgias following succinylcholine. PMID- 8703459 TI - Potency of doxacurium in infants, children, and adolescents during N2O-O2 alfentanil anesthesia. AB - STUDY OBJECTIVE: To evaluate neuromuscular potency of doxacurium during balanced anesthesia in pediatric patients. DESIGN: Prospective, consecutive sample trial. SETTING: Operating room at a university hospital. PATIENTS: 15 infants (1 to 11 months), 20 children (3 to 10 years), and 20 adolescents (13 to 19 years). INTERVENTIONS: Anesthesia was induced and maintained with thiopental, alfentanil, and nitrous oxide in oxygen. No volatile drugs were used at any time during the study. The neuromuscular function was recorded as adductor pollicis electromyography evoked by a train-of-four stimulation at 20-second intervals. A cumulative log-dose probit-response curve of doxacurium was established for every patient. MEASUREMENTS AND MAIN RESULTS: ED50 and ED95 doses of doxacurium (14 micrograms/kg and 25 micrograms/kg in infants, 26 micrograms/kg and 53 micrograms/kg in children, and 20 micrograms/kg and 41 micrograms/kg in adolescents, respectively) were smallest in infants and greatest in children (p < 0.05 between each pair of groups by analysis of variance and Scheffe's F-test). CONCLUSIONS: Potency of doxacurium was greatest in infants and least in children. We suggest that doxacurium can be administered safely in infants, and with dosages close to those reported in adults. Children's dose requirement was almost 50% greater than that of infants. PMID- 8703460 TI - Evaluation of laboratory coagulation and lytic parameters resulting from autologous whole blood transfusion during primary aortocoronary artery bypass grafting. AB - STUDY OBJECTIVE: To determine if autologous blood reinfusion influences overall hemostatic function following aortocoronary artery bypass graft (CABG) surgery, and if so, where the predominant area of this influence lies. DESIGN: Prospective, with control values on each patient. SETTING: Cardiac operating room of a major university-affiliated county hospital. PATIENTS: 20 patients undergoing elective CABG surgery. INTERVENTIONS: Following heparinization, and prior to cardiopulmonary bypass (CPB), venous blood (average 4.9 ml/kg) was removed via an indwelling internal jugular catheter into a preservative-free plastic transfer pack unit and stored without agitation at room temperature. This autologous whole blood was reinfused after systemic protamine reversal of heparin. Blood samples for analysis were drawn immediately before and 5 minutes after completion of the reinfusion. MEASUREMENTS AND MAIN RESULTS: Autologous blood reinfusion appears to be significantly related to increased hemoglobin, hematocrit, platelet count, fibrinogen, plasminogen, and antiplasmin levels. The prothrombin time and activated partial thromboplastin times decreased significantly, whereas activated clotting times and D-dimer levels were unchanged. Significant increases occurred in the following thromboelastography parameters: maximum amplitude, amplitude 60 minutes after the maximum amplitude, and whole blood clot lysis index. Reaction time and coagulation time were not statistically different from control values. CONCLUSIONS: Significant improvements in coagulation and lytic parameters occur following CPB after the infusion of autologous blood. These improvements in coagulation indices may be the result of the infused blood or hemoconcentration, which is also known to occur during this period. Additional control studies are needed to differentiate these effects. PMID- 8703461 TI - Effects of Trendelenburg and reverse Trendelenburg postures on lung and chest wall mechanics. AB - STUDY OBJECTIVE: To test whether the Trendelenburg ("head-down") or reverse Trendelenburg ("head-up") postures change lung and chest wall mechanical properties in a clinical condition. DESIGN: Unblinded study, each patient serving as own control. SETTING: University of Maryland at Baltimore Hospital, Baltimore, Maryland. PATIENTS: 15 patients scheduled for laparoscopic surgery. INTERVENTIONS: Patients were anesthetized and paralyzed, tracheally intubated and mechanically ventilated at 10 to 30 per minute and at a tidal volume of 250 to 800 ml. Measurements were made before surgery in supine, head-up (10 degrees from horizontal) and head-down (15 degrees from horizontal) postures. MEASUREMENTS AND MAIN RESULTS: Airway flow and airway and esophageal pressures were measured. From these measurements, discrete Fourier transformation was used to calculate elastances and resistances of the total respiratory system, lungs, and chest wall. Total respiratory elastance and resistance increased in the head-down posture compared with supine due to increases in lung elastance and resistance (p < 0.05); but chest wall elastance and resistance did not change (p > 0.05). Lung elastance also exhibited a negative dependence on tidal volume while head-down that was not observed in the supine posture. The change in lung elastance compared with supine was positively correlated to body mass index (weight/height2) and negatively correlated to tidal volume. Lung and chest wall elastance and resistance were not affected by shifting from supine to head-up (p > 0.05). CONCLUSIONS: The Trendelenburg posture increases the mechanical impedance of the lung to inflation, probably due to decreases in lung volume. This effect may become clinically relevant in patients predisposed with lung disease and in obese patients. PMID- 8703462 TI - Ketorolac and propofol anesthesia in a patient taking chronic monoamine oxidase inhibitors. AB - There is continued controversy regarding the concurrent use of monoamine oxidase inhibitors (MAOIs) and opioids in anesthesia. MAOIs are being used in both psychiatry and also in the treatment of Parkinson's disease. The adverse interaction of meperidine with MAOIs is well described in the literature. Several other opioids used in anesthesia have been reported as potentially safe and also implicated in interaction with MAOIs. We present a case report using ketorolac as a nonopioid analgesic during a general anesthesia combined with midazolam and propofol in a successful approach in a patient continuing MAOI therapy. PMID- 8703463 TI - A complication of combined regional anesthesia techniques for ophthalmic surgery. PMID- 8703464 TI - Chest wall rigidity during infusion of fentanyl in a two-month-old infant after heart surgery. PMID- 8703465 TI - Anesthetic implications of Larsen syndrome. AB - In this article I present two children with Larsen syndrome who required anesthetic care during surgical procedures of the ear-nose-throat (ENT) region. The anesthetic implications of this disorder are discussed. Larsen syndrome consists of multiple joint dislocations and a characteristic facies. Associated orthopedic abnormalities include cervical spine vertebral body hypoplasia and hemivertebrae, which may lead to cervical kyphosis and compromise of cervical cord function. Of prime importance to the anesthesiologist are the associated defects of the airway including laryngotracheomalacia, which may predispose the patient to perioperative respiratory compromise. Associated cardiac defects have also been described including atrial and ventricular septal defects. PMID- 8703466 TI - Simultaneous oral and nasal tracheal intubation utilizing a fiberoptic scope in a patient with facial trauma. AB - We report a case of an oral intubation in a facial trauma patient that required exchanging the oral tube to a nasal endotracheal tube. This was accomplished by utilizing the fiberoptic bronchoscope to introduce a nasal tracheal tube beside the oral tube prior to removal of the oral tube from the trachea. This allowed continuation of oxygenation and ventilation until the airway was subsequently secured with the nasal tracheal tube. This approach may help avoid the loss of the airway and subsequent intervention with a surgical airway under poor conditions. PMID- 8703467 TI - Potential risk associated with anesthesia machines with single oxygen cylinders. PMID- 8703468 TI - Laryngospasm associated with use of a passive humidifier with the laryngeal mask airway. PMID- 8703469 TI - A method and apparatus for testing the anesthesia machine circuit unidirectional values. PMID- 8703470 TI - Inactivation of tumor suppressor proteins in lung cancer. AB - It had been thought that the central molecular event in the malignant transformation of a cell is the mutation of certain oncogenes-and the resultant dysregulated activation of their encoded proteins. During the past decade, however, it has become clear that alteration of the activity of the protein products of tumor suppressor genes, through mutation or at the posttranslational level, is an equally basic and universal process in tumorigenesis. These proteins normally modulate cellular proliferation in the developing and adult organism, functioning as tumor suppressors by inhibiting inappropriate cell division. Therefore, inactivation of the normal function of tumor suppressor proteins removes important regulatory constraints on the cell, permitting the accelerated growth of cancerous tissue. The genesis of lung cancer is though to involve between 10 and 20 mutations. Of these, several are now known to involve tumor suppressor genes. In this review I will discuss the mechanism of tumor suppression by the protein encoded by one of these, the retinoblastoma gene, to illustrate precisely why the inactivation of tumor suppressors is a requisite step in cellular progression to lung and other carcinomas. PMID- 8703472 TI - Expression of Hoxb genes in the developing mouse foregut and lung. AB - Lung development in the mouse begins at embryonic day 9.5 (E9.5) when lung buds form in the foregut. Subsequently, there is extensive branching and cellular differentiation that depends upon specific epithelial-mesenchymal interactions. Homeobox genes are expressed in specific temporo-spatial patterns in the developing embryo and are known to be involved in axial patterning and specification of regional identity. Using whole mount in situ hybridization and immunohistochemistry, we studied the expression of Hoxb-1, b-2, b-3, b-4 and b-5 in the E9.5-E14.5 foreguts and lungs. Our results show that in E9.5 branchial arches and foregut, Hoxb genes are expressed in overlapping spatial domains and the anterior boundaries of these domains correspond to the position of a particular gene in the cluster-genes on the 3' end of the cluster are expressed more anteriorly in the branchial arches and foregut and those on the 5' end are expressed more posteriorly. Three of the genes, Hoxb-3, b-4, and b-5, are highly expressed in the foregut where the lung buds form. In contrast, in E10.5-E14.5 lung, there are two patterns of Hoxb gene expression. Hoxb-3 and b-4 are expressed in the mesenchyme of the trachea, mainstem bronchi, and distal lung, whereas Hoxb-2 and b-5 mRNA are present only in the mesenchyme of the distal lung buds. These results suggest that specific combinations of Hoxb gene expression are important in lung development and that Hoxb genes may be involved in specifying the differences between proximal (trachea and main bronchi) and distal (lung bud) mesenchyme. PMID- 8703471 TI - Hox genes in the lung. PMID- 8703474 TI - Differential expression of laminins and their integrin receptors in developing and adult human lung. AB - Laminins (Ln) appear to play an important role in the morphogenesis of airways. We studied the expression of different laminin chains and their integrin receptors in fetal and adult lung by immunohistochemistry. Special attention was focused on the changes in the expression of these proteins during the development from the pseudoglandular (PG) and canalicular stages to adult lung, and on the possible implications of the changes for the normal lung development. The most significant changes in the expression pattern were found during the development from the PG stage to the canalicular stage. Basement membranes (BM) of both the epithelial buds and the becoming bronchi showed reactivity for Ln-alpha 1, -alpha 3, and -beta 3 chains at all stages. The alpha 2 chain was expressed only in the epithelial buds at the PG stage, and could not be found in any epithelial structures at the canalicular stage. Similarly, at the PG stage the Ln-beta 2 chain was expressed in BMs of both epithelial buds and bronchi but disappeared from the bronchial BM before the canalicular stage. Ln-beta 1 chain appeared in the bronchial BM first in the mature lung, which suggests the presence of uncharacterized Ln-beta chains earlier in development. There were considerable changes in the expression of integrins (Int) concomitantly with alterations in the composition of the BMs. At the PG stage the epithelial buds showed reactivity for Int-alpha 2 -alpha 3, and -alpha 6 subunits, but at the canalicular stage the Int-alpha 2 and -alpha 6 subunits disappeared, and only Int-alpha 3 integrin subunit was found in evolving alveolar walls; Int-alpha 6 was found in capillaries. A similar distribution of Int subunits was also found in adult alveoli. The bronchi expressed Int-alpha 2, -alpha 3, and -alpha 6 subunits at all developmental stages, but the Int-beta 4 subunit emerged first at the canalicular stage. Our results suggest that there are major changes in the expression of Ln and their Int receptors during morphogenesis of the lung, which may be important for normal development. PMID- 8703473 TI - Role of very late activation antigen-4 in the antigen-induced accumulation of eosinophils and lymphocytes in the lungs and airway lumen of sensitized brown Norway rats. AB - We used flow cytometry and treatment in vivo with a monoclonal antibody (mAb), TA 2, to the alpha 4 integrin to investigate the role of alpha 4 beta 1, CD49d/CD29 (VLA-4) in antigen-induced lung inflammation in Brown Norway (BN) rats. Ovalbumin (OVA) inhalation induced an accumulation of eosinophils and lymphocytes in the lungs and bronchoalveolar lavage (BAL) fluid of sensitized BN rats at 24 h after challenge. Phenotypic analyses demonstrated that the percentages of T cells expressing detectable alpha 4 and CD25 in the bronchial lumen after antigen challenge were dramatically increased compared with blood and lymph node T cells. The mean channel fluorescence values of alpha 4 expression were also increased on BAL T cells compared with blood or lymph node T cells. Treatment of OVA sensitized rats in vivo with total cumulative doses of 0.75 to 6 mg/kg TA-2 mAb intraperitoneally produced dose-related increases in circulating TA-2 and a peripheral blood lymphocytosis, basophilia, and eosinophilia. Flow cytometric analysis of the peripheral blood T cells after in vivo TA-2 mAb administration showed decreases in detectable alpha 4 when these cells were examined ex vivo. Treatment with TA-2, but not an isotype-matched control mouse immunoglobulin G1 mAb, markedly inhibited the OVA-induced recruitment of lymphocytes and eosinophils into the airway lumen. Very few CD3+CD49d+ cells migrated into BAL fluid following anti-alpha 4 mAb treatment in vivo. Treatment with TA-2 also significantly attenuated OVA-induced inflammatory histopathology. We conclude that VLA-4 is a critically important adhesion molecule involved in antigen specific lung inflammation in sensitized BN rats. PMID- 8703475 TI - Nonadhesive stationary organ culture of human bronchial mucosa. AB - The supply of fresh bronchial tissue from human donors for in vitro culture is limited. Routine fiberoptic bronchoscopy offers a safe and easy procedure for obtaining minor biopsies and we wanted to see if the material provided could be used for organ culture by using a simple liquid overlay technique. Bronchial biopsies were cut into fragments 400-500 microns and kept immersed in a standard serum-supplemented medium for 40 days. An agar base prevented adhesion of the tissue. By light and electron microscopy it was shown that the tissue fragments had a differentiated epithelium at their surface throughout the culture period. An outgrowth of epithelial cells on the scaffold of the exposed stroma, covering the surface of the whole fragment, occurred within the first 5 days of culture. This epithelium was partly ciliated, 2-4 cell layers thick with squamous and cuboidal cells and expressed epithelial markers (cytokeratin and Ber-Ep4). The amount of cilia increased during the first 15 days of culture. The epithelium rested on a neosynthesized basement membrane as visualized by electron microscopy and immunohistochemistry with antibodies directed against collagen IV, laminin, and fibronectin. The central stroma consisted of loose connective tissue with fibroblasts. This simple tissue culture model combines maintenance and neoformation of bronchial epithelium on top of a living natural substrate, thus enabling direct biological studies on clinical biopsy material under perfectly viable conditions. PMID- 8703476 TI - Characterization of a neutrophil inhibitor peptide harvested from human bronchial lavage: homology to influenza A nucleoprotein. AB - Bronchi are exposed to particulate matter, including bacteria, fungi and dusts, that should trigger release of molecules which attract polymorphonuclear neutrophils (PMN). However, normal bronchi are relatively devoid of PMN, suggesting that there exists a mechanism to dampen acute inflammation in the lung. We have previously reported that bronchial lavage from normal humans contains a nonpolar peptide that inhibits PMN chemotaxis and oxidant production. In the present study we devised preparative methods to obtain sufficient quantities of a similar inhibitor molecule for partial amino acid sequencing and allow production of truncated analogues. Amino acid sequencing demonstrated that the peptide includes a 10-amino-acid sequence that is completely homologous to a sequence of amino acids contained in the influenza A nucleoprotein. Synthesized peptides containing this 10-amino-acid sequence inhibited PMN chemotaxis and oxidant production. In addition, PMN lysates actively phosphorylated peptides containing the 10-amino-acid sequence or a partial sequence containing an apparent phosphorylation site. U937 cells were noted to be one source of this inhibitor, as a similarly sized nonpolar inhibitor peptide was purified from U937 culture supernatants. In addition, U937 and monocyte cellular lysates contained proteins recognized by an antiserum directed at the influenza A nucleoprotein. Further characterization of the molecule described in this study or related molecules may lead to significantly new antiinflammatory strategies. PMID- 8703478 TI - Neutrophil transepithelial migration is dependent upon epithelial characteristics. AB - To better understand the mechanisms by which neutrophils migrate to the airway lumen during an inflammatory response, we constructed an in vitro model system to examine the interactions of human neutrophils, human lung epithelial cells, mediators, and proinflammatory cytokines. We directly compared neutrophil movement through three lung epithelial cell lines, A549, H441, and 16-HBE-14o, in response to three chemoattractants, FMLP, LTB4, and IL-8, and the proinflammatory cytokines IL-1 alpha and beta and TNF alpha. While there was variation in the responses to the chemotaxins, there was no correlation between the transmonolayer electrical resistance and the ability of the neutrophils to migrate across the epithelia in response to the agents used. FMLP, IL-8, and LTB4 induced dose- and time-dependent neutrophil migration across all three epithelia. However, TNF alpha- and IL-1-induced neutrophil migration occurred only through monolayers that produced soluble chemoattractants in response to these cytokines. Although all three epithelia produced low amounts of IL-8 constitutively, the capacity of IL-1 and TNF alpha to induce transepithelial migration was directly associated with the ability of the epithelia to produce large amounts of IL-8 in response to IL-1 and TNF alpha. We conclude that the phenotype of the epithelial cell (e.g., capacity to produce IL-8) affects stimulated neutrophil transepithelial migration. PMID- 8703477 TI - Detection of N7-methyldeoxyguanosine adducts in human pulmonary alveolar cells. AB - Alkylating agents may cause DNA damage in different human cells and tissues, including lungs. For instance, tobacco-specific N-nitrosamines are known to produce methyl-DNA adducts, such as N7-methyldeoxyguanosine, and to induce lung tumors. We applied a combined high-performance liquid chromatography (HPLC)/32P postlabeling technique for measurement of N7-methyldeoxyguanosine in human pulmonary alveolar cells (HPAC). Thirty patients (13 males, 17 females; mean age 51 +/- 17 yr) undergoing bronchoalveolar lavage for diagnosis of nonmalignant lung diseases were studied. DNA was extracted from HPAC, digested to 2' deoxyribonucleotide 3'-monophosphates and HPLC separated to obtain deoxyguanosine (dGp) and N7-methyldeoxyguanosine (N7-MedGp) monophosphates. Fractions corresponding to normal (1:10,000) and N7-methylated dGp were subsequently 32P postlabeled by T4 polynucleotide kinase with high specific activity 32P-ATP, resolved by two-dimensional thin-layer chromatography (TLC) and autoradiographed after 3 to 18 h exposure. Spots corresponding to dGp and N7-MedGp were scraped off the plates and quantitated by liquid scintillation counting to calculate direct molar ratios. Recovered HPAC (14.4 +/- 10.0 x 10(6)) were predominantly macrophages (73.8 +/- 16.4%) and lymphocytes (9.8 +/- 11.6%). N7-MedGp was detected in 11 patients, the level ranging from 0.10 to 48.03 fmol/micrograms DNA which corresponded to 0.31-79.00 x 10(-6) N7-MedGp/dGp ratios. Detection of N7 MedGp in HPAC was associated with the smoking habit of patients: N7-MedGp was present in 7 of 10 smokers, 2 of 10 ex-smokers, and 2 of 10 nonsmokers (P < 0.05). These results show that HPAC may be used for molecular dosimetry of DNA damage by alkylating agents, including tobacco-specific N-nitrosamines, in cigarette smokers and thus used for cancer risk assessment. PMID- 8703479 TI - Pulmonary neuroendocrine cells in cultures of human infant airway mucosa from postmortem tissues. AB - Airway mucosa was isolated by enzymatic dissociation from tracheas and bronchi obtained at autopsy from stillbirth, infants, and children. An epithelial cell fraction was recovered that could be cultured on a collagen-coated substratum for at least 7 days. Epithelial cells were identified immunocytochemically with anticytokeratin antibody; pulmonary neuroendocrine cells (PNEC) were identified with antibodies against serotonin, bombesin/GRP, and MOC-1 cell surface antigen. Transmission electron microscopy confirmed neuroendocrine ultrastructural features of PNEC including the presence of neurosecretory dense core vesicles. The ability to recover and maintain viable PNEC in human lung airway epithelial cell cultures from postmortem tissues should facilitate further investigations of PNEC function in normal human lung and in various disease states. PMID- 8703481 TI - Corticosteroids induce intracellular interleukin-1 receptor antagonist type I expression by a human airway epithelial cell line. AB - Interleukin-1 (IL-1) is an important proinflammatory cytokine which may contribute to the pathogenesis of inflammatory airway disorders, such as asthma and cystic fibrosis. Interleukin-1 receptor antagonist (IL-1ra) is a naturally occurring IL-1 inhibitor which binds to IL-1 receptors without inducing agonist activity. Three IL-1ra isoforms have been identified: secreted IL-1ra (sIL-1ra), which is preferentially expressed by-inflammatory cells; intracellular IL-1ra (iIL-1ra) type I, which lacks a signal peptide and is preferentially expressed by epithelial cells; and iIL-1ra type II, which is identical to iIL-1ra type I except for the insertion of an additional 21 amino acids. The goal of this study was to assess whether airway epithelial cell iIL-1ra type I production can be regulated by corticosteroids. First, using reverse transcription-polymerase chain reaction (RT-PCR) and immunoblotting, we confirm that normal human bronchial epithelial (NHBE) cells and a human pulmonary mucoepidermoid carcinoma cell line (NCI-H292) express intracellular IL-1ra type I messenger RNA (mRNA) and protein. Second, using immunoblotting and ELISA, we report that dexamethasone induces time and concentration-dependent increases in iIL-1ra type I protein within NCI-H292 cell lysates. Lastly, utilizing a ribonuclease protection assay, we report that dexamethasone induces concentration-dependent increases in iIL-1ra type I mRNA levels in NCI-H292 cells. These data suggest that corticosteroid-mediated induction of iIL-1ra type I mRNA and protein by human bronchial epithelial cells represents a novel mechanism by which IL-1-mediated airway inflammatory events might be regulated. PMID- 8703480 TI - Expression of MUC1 mucin gene by hamster tracheal surface epithelial cells in primary culture. AB - Primary hamster tracheal surface epithelial (HTSE) cells carry mucin-like glycoproteins on the apical surface which are releasable by neutrophil elastase. In some cancer cells, mucins are localized on the cell surface and have been shown to be encoded by the MUC1 mucin gene. The objectives of the present experiments were: (I) to determine if HTSE cells express MUC1 mucin gene; (2) if they do, to isolate and characterize the hamster MUC1 complementary DNA (cDNA); and (3) to examine the pattern of MUC1 mRNA expression at different stages of culture. Reverse transcriptase-polymerase chain reaction amplification of HTSE cell RNAs using degenerate primers based on homologous sequences between the human and mouse MUC1 genes revealed the presence of a cDNA (0.5 kb) which has an 88% similarity in sequence with the mouse MUC1 cDNA. Using this 0.5 kb cDNA as a probe, an HTSE cell cDNA library was screened to isolate a hamster MUC1 cDNA clone. Sequence analysis of the cDNA revealed that it encodes an integral membrane protein of 676 amino acids which consists of (1) an N-terminal signal sequence, (2) the tandem repeat domain encoding 12 repeats of 20 amino acids, and (3) the C-terminal region consisting of degenerate tandem repeats and a unique sequence containing both the transmembrane and cytoplasmic domains. The presence of seven tyrosine residues in the cytoplasmic domain suggests a potential role as a receptor. Finally, expression of MUC1 mucin gene in HTSE cells appears to be associated with differentiation of secretory cells. PMID- 8703482 TI - Plasmin regulates the activation of cell-associated latent TGF-beta 1 secreted by rat alveolar macrophages after in vivo bleomycin injury. AB - Transforming growth factor beta s (TGF-beta s) are 25-kD multifunctional proteins that regulate inflammation and connective tissue synthesis. With rare exception TGF-beta 1 is secreted noncovalently bound to a latency-associated peptide (LAP) that renders the mature TGF-beta 1 biologically inactive. An important mechanism for the control of TGF-beta 1 action is the regulation of the post-translational processing that removes the LAP from the mature peptide and renders it biologically active. In a model of pulmonary inflammation and fibrosis induced by the antineoplastic antibiotic, bleomycin, we have demonstrated that explanted alveolar macrophages secrete progressively increasing quantities of a biologically active form of TGF-beta 1, the secretion of which was maximal 7 days after bleomycin administration. Thereafter, there was a rapid decline in the secretion of the active form of TGF-beta 1, whereas the latent form continued to be secreted in elevated quantities. Plasmin, a serine protease, was transiently generated by the same bleomycin-activated alveolar macrophages and paralleled the rise in active TGF-beta 1. When alpha 2-antiplasmin, an inhibitor of plasmin, was added to cultures of alveolar macrophages, the post-translational activation of L TGF-beta 1, was totally abrogated. When plasmin was added to alveolar macrophages in culture, there was complete activation of the L-TGF-beta 1 that had been secreted during the culture period. However, there was no effect of plasmin on the same alveolar macrophage-derived L-TGF-beta 1 in cell-free conditioned media. Our findings suggest that the secretion of an active form of TGF-beta 1 by alveolar macrophages is regulated by the generation of plasmin and requires that the alveolar macrophages be present. Because the diminution of active TGF-beta 1 coincides with the resolution of inflammation, this suggests that the availability of plasmin regulates the biologically active form of TGF-beta 1, and thus, the inflammation seen after bleomycin-induced lung injury. PMID- 8703483 TI - Role of Leukotriene-degrading enzymes in pulmonary response to antigen infusion in sensitized guinea pigs in vivo. AB - To determine the role of leukotriene (LT)-degrading enzymes in allergic reactions, we studied the effects of inhibitors of gamma-glutamyl transpeptidase (gamma-GTP) and dipeptidases on increases in pulmonary insufflation pressure (PIP) and vascular permeability induced by ovalbumin (OA) antigen in guinea pigs sensitized to OA antigen in vivo. Vascular permeability was assessed by the amount of extravasated Evans blue dye from the trachea, main bronchi, and segmental bronchi. An intravenous (i.v.) administration of OA antigen (200 micrograms/kg) caused increases in PIP and extravasated Evans blue dye, and OA antigen-induced effects were potentiated by gamma-GTP inhibitor L-serine borate (3 x 10(-5) M/kg, i.v.) (P < 0.05) and an inhibitor of dipeptidases, L-cysteine (3 x 10(-5) M/kg, i.v.) (P < 0.01). OA antigen-induced increases in PIP and Evans blue dye extravasation were in part inhibited by LT-receptor antagonist ONO-1078 (10(-4) M/kg, i.v.). Guinea-pig tracheal tissues contained gamma-GTP and microsomal dipeptidase activities. Histochemical and immunohistochemical studies indicate that gamma-GTP-like activity existed in the epithelium and smooth muscle, and an activity of microsomal dipeptidase was observed in the endothelial cells of microvessels and epithelium. These results suggest that LT-degrading enzymes have an important role in regulating allergic reaction in the airway in vivo. PMID- 8703484 TI - Time course of chemotactic factor generation and neutrophil recruitment in the lungs of dust-exposed rats. AB - The time course of neutrophil recruitment into the lung, neutrophilic chemotactic activity, and the gene expression of neutrophilic chemokines by lavaged cells was determined after intratracheal instillation of various particles. Low-toxicity, low-solubility dusts such as titanium dioxide (TiO2) particles, as well as fibrogenic crystalline silica and nonfibrogenic amorphous silica particles were instilled into the lungs of rats. Results showed that all three dusts induced neutrophilic inflammation as early as 5 h after exposure. Both crystalline and amorphous silica elicited higher degrees of pulmonary inflammation when compared with TiO2 particles. Maximal infiltration of neutrophils into the lungs occurred 5 to 6 h after intratracheal instillation of the dusts. The inflammatory response was transient for TiO2 and amorphous silica, i.e., evident at 2 days after exposure but not different from controls at 10 days after exposure. In contrast, inflammatory effects were sustained through a 10-day period following exposures to crystalline silica. Chemotactic activity for neutrophils was detected directly in bronchoalveolar lavage (BAL) fluids of dust-exposed rats within 2 h after exposure, but not in the BAL fluids of saline- or unexposed rats. The chemotactic activity was correlated with the influx and disappearance of neutrophils into alveolar regions of the lung in TiO2- and amorphous silica-exposed rats. The mRNA expression of two known neutrophil chemotactic cytokines in BAL cells, macrophage inflammatory protein-2 (MIP-2) and KC, also correlated with chemotactic activity and acute and pulmonary inflammatory responses. MIP-2 mRNA was expressed prior to the detection of chemotactic activity in BAL fluids. However, the mRNA expressions of MIP-2 and KC were transient for rats that were exposed to these dusts as KC and MIP-2 message were no longer detectable in BAL cells after 2 days of recovery. Although both neutrophilic chemotactic activity and inflammation remained prominent 10 days after exposure to crystalline silica, MIP-2 expression could not be detected in BAL cells. Thus, we conclude that MIP-2 is likely to be only one of several cytokines involved in mediating neutrophilic inflammation following a single instillation of crystalline silica. PMID- 8703485 TI - 4-Hydroxynonenal mimics ozone-induced modulation of macrophage function ex vivo. AB - Ozone is a ubiquitous pollutant that can cause acute pulmonary inflammation, cellular injury and may contribute to the development or exacerbation of chronic lung diseases. Despite much research, the effects of ozone on humans and potential cellular mechanisms of injury are still uncertain. However, ozone has been reported to increase the formation of aldehydes that could react with cellular proteins. Therefore, the purpose of these studies was to determine whether 4-hydroxynonenal (HNE), a previously unidentified aldehyde product of ozone exposure, is formed in human subjects exposed to ozone, and whether the response of human alveolar macrophages (AM) following a 1-h exposure to 0.25 ppm ozone with moderate exercise could be mimicked by in vitro incubation of AM with HNE. Western analysis demonstrated increased HNE protein adducts in airway fluid and alveolar macrophages after ozone exposure. AM were examined for endotoxin (lipopolysaccharide [LPS])-stimulated interleukin-1 beta (IL-1 beta) release and expression of heat shock protein 72 (HSP72). Immediately after ozone exposure there was no change in HSP72, but a 5-fold increase occurred 4 h after exposure. By 18 h after exposure, HSP72 levels decreased to below comparable air-exposed levels. Immediately after ozone exposure there was no effect on IL-1 beta release stimulated by LPS. However, IL-1 beta release stimulated by LPS was significantly inhibited 4 h after ozone exposure. By 18 h after ozone exposure, IL-1 beta release stimulated by LPS returned to normal. Incubation of human AM in vitro with HNE induced HSP72 and blocked LPS-stimulated IL-1 beta release possibly by inhibiting interleukin converting enzyme. Consequently, the in vitro results and demonstration of HNE protein adducts following ozone exposure are consistent with HNE being involved in this process in vivo and suggest that the cellular toxic effects of ozone could be a result of thiol reactive aldehydes produced by ozone. PMID- 8703486 TI - Alginate, the slime exopolysaccharide of Pseudomonas aeruginosa, binds human leukocyte elastase, retards inhibition by alpha 1-proteinase inhibitor, and accelerates inhibition by secretory leukoprotease inhibitor. AB - The interaction of alginate from Pseudomonas aeruginosa ATCC 39324 with human leukocyte elastase was studied by determining the effects of the polysaccharide on the amidolytic activity of the enzyme toward a range of synthetic peptide substrates of different length. The data support a model in which each elastase molecule interacts with a total of 19 uronic acid units on the alginate, primarily through electrostatic forces. Binding of alginate results in occlusion of distal subsites, most likely S4 and S5, of the enzyme's extended substrate binding domain. As a result, alginate alone appears to be a weak inhibitor of the hydrolysis of long synthetic peptide substrates and [14C]elastin by elastase. Alginate also has effects on the antielastase function of naturally occurring protease inhibitors in the lung: It reduces the association rate of elastase and alpha 1-proteinase inhibitor, whereas it increases the association rate of elastase and secretory leukoprotease inhibitor. In the presence of 36 micrograms/ml alginate, the median concentration found in sputum from cystic fibrosis patients infected with mucoid strains of P. aeruginosa, the second-order rate constant for inhibition of elastase by secretory leukoprotease inhibitor is 2.6-fold greater than that for alpha 1-proteinase inhibitor. Alginate has only a minor effect on the antielastase activities of elafin and a recombinant form of the isolated C-terminal domain of secretory leukoprotease inhibitor. Based on these findings, alginate may be an important factor in determining the local distribution of leukocyte elastase and perturbing the overall protease antiprotease balance in the infected lungs of cystic fibrosis patients. PMID- 8703487 TI - Effect of cardiomyoplasty on systolic and diastolic function. AB - Although cardiomyoplasty has become a recognized treatment for end-stage heart failure, the effects of this procedure on systolic and diastolic function are still unclear. To determine the effects of paced and non-paced latissimus dorsi cardiomyoplasty on systolic and diastolic function, the maximal elastance of the left ventricle (Emax), stroke volume, preload recruitable stroke work and diastolic compliance were measured in an experimental heart failure model. Collateral blood vessels to the latissimus dorsi were ligated 2 weeks before cardiomyoplasty in order to reduce the risk of ischemic injury. Histological examination of muscle biopsies confirmed that the two-stage procedure preserved normal muscle architecture. The non-paced cardiomyoplasty wrap adversely affected both systolic and diastolic function. Paced Latissimus Dorsi during heart failure improved systolic function but had no measurable effect on diastolic function. CONCLUSIONS: 1. Non-paced, or unstimulated, latissimus dorsi cardiomyoplasty acutely impairs cardiac function. 2. Delayed cardiomyoplasty, 2 weeks after collateral ligation, prevents ischemic injury to the muscle flap. PMID- 8703488 TI - Simultaneous coronary bypass and abdominal aortic surgery in patients with severe coronary disease--indication and results. AB - In patients with severe coronary artery disease (CAD) abdominal aortic surgery is still associated with high morbidity and mortality rates. Some patients will present with both symptomatic CAD and large, symptomatic abdominal aortic aneurysms (AAA) or end-stage aortic occlusive disease (AOD) that does not allow for a two-stage procedure. We report a series of 29 patients who underwent simultaneous coronary artery bypass graft surgery (CABG) and abdominal aortic surgery (25 AAA, 4 AOD). In the AAA group there were 23 males and 2 females with a mean age of 68 years (50-80). Sixteen patients presented with severe three vessel disease. Ten patients had unstable angina. Aortic stenosis or insufficiency was present in two and one patient, respectively. Four patients with three-vessel disease and an ejection fraction below 30% presented with end stage AOD and critical limb ischemia. Coronary bypass graft surgery was performed first. With the patient still on partial cardiopulmonary bypass, abdominal aortic surgery was carried out. Patients received an average of 3.1 coronary bypass grafts. Additionally, three aortic valves were implanted. Fourteen tube grafts and 15 bi-iliacal or bifemoral bifurcation grafts were placed in the abdominal aortic position. Additional vascular surgery was performed in five patients. Intraoperative management was without complication in all but one patient, who had intraoperative myocardial infarction (AOD group). Hospital mortality was 8% (2/25) in the AAA group. There was however substantial hospital morbidity (52.2%). The mean follow-up is 20.5 +/- 2.5 months. The actuarial survival rate at 3 years is 84.9%. It is concluded that combined CABG and abdominal aortic surgery is a reasonable option for patients who present with both severe CAD and symptomatic abdominal aortic disease. The continuation of CPB during aortic surgery may effectively prevent the adverse effects of infrarenal aortic clamping on a failing ventricle. PMID- 8703489 TI - Mortality during the two years after coronary artery bypass grafting in relation to perioperative factors and urgency of operation. AB - The purpose of this study was to describe mortality during the 2-year-period after coronary artery bypass surgery (CABG) in relation to perioperative risk factors and urgency of operation. All the patients in western Sweden were included in whom CABG was performed between June 1988 and June 1991, without concomitant procedures or re-operations. The study was prospective in design. In all, 2000 patients were operated upon and 186 (9.3%) of the operations were acute. There was a significant relationship between the urgency of the operation and mortality. Early mortality was 2.4% in elective operations and 5.4-62.5% in urgent to emergency operations. The 30-day to 2-year mortality was 4.2%. The perioperative risk indicators independently associated with early mortality were neurologic complications, serum-aspartate aminotransferase (S-ASAT) more than 2.0 microkat/l, urgency of operation, the use of circulatory assist devices, re operation and ventilator time more than 24 h. The risk indicators for mortality after 30 days were pneumothorax, longer intensive care unit (ICU) time, the use of inotropic drugs and neurologic complications. In conclusion, the multivariate analysis reveals the urgency of the operation as a predictor of early mortality after CABG, but no significant association with mortality was found after 30 days. When excluding death within 30 days, three additional independent predictors of mortality were identified. PMID- 8703490 TI - Comparative study of the hydrodynamic function of six size 19 mm bileaflet heart valves. AB - The hydrodynamic function of six size 19 mm bileaflet mechanical heart valves was investigated in a pulsatile flow simulator. The St. Jude Standard, St. Jude Haemodynamic Plus, CarboMedics, Sorin Biomedica, Duromedics and ATS valves were studied. The mean pressure drop and effective orifice area during forward flow, regurgitant volumes and total energy loss were analysed. The St Jude Haemodynamic Plus and Sorin Biomedica valves had significantly lower forward flow pressure drops and total energy losses than the other four valves. Assuming that all six valves can be implanted in the equivalent size of aortic root, this should lead to improved in vivo performance. PMID- 8703491 TI - Medium-term results after surgery for aortic arch aneurysm with hypothermic cerebral perfusion. AB - We have used two techniques of hypothermic cerebral perfusion (CP) for the surgical treatment of aortic arch aneurysm in the last 10 years. Between March 1985 and December 1993, 83 patients underwent surgery for aortic arch aneurysm. Fifty-one cases had aortic dissection (AD) in the transverse arch and/or its branches, and 32 cases showed true aneurysm (TA) of the aortic arch. In those 83 patients, 37 cases received antegrade CP and 46 cases underwent retrograde CP. Surgical results were compared among the groups by Kaplan-Meier actuarial method and Cox-Mantel statistical analysis. The early mortality after surgery for aortic arch aneurysm was 11.8% in the AD group and 21.9% in the TA group. The early mortality was 21.6% with antegrade CP and 10.9% with retrograde CP. The 6-year actuarial survival rate was 71.7% in the TA group and 67.1% in the AD group. In the AD group, the 3-year survival rate was 93.9% with retrograde CP and 61.1% with antegrade CP (P < 0.005). In the TA group, the 3-year survival rate was similar for antegrade CP (73.3%) and retrograde CP (69.2%). These results suggest that current surgical results of aortic arch aneurysm with hypothermic CP are acceptable and the retrograde CP technique might be recommended, especially for surgery of aortic arch aneurysm with AD. PMID- 8703492 TI - Myocardial protection with and without leukocyte depletion: a comparative study on the oxidative stress. AB - We tested the hypothesis that controlled reperfusion with leukocyte-depleted blood could improve myocardial protection by reducing the oxidative stress in patients undergoing myocardial revascularization. Thirty-four patients receiving antegrade/retrograde blood cardioplegia were divided into: group A: 11 patients with ejection fractions (EF) less than 35%, treated with leukocyte-depleted controlled blood reperfusion, group B: 11 patients with EF less than 35% in whom no leukocyte depletion was performed, group C: 6 patients with EF more than 45% treated as group A and group D: 6 patients with EF more than 45% without leukocyte depletion. To asses the oxidative stress, we evaluated total, total oxidized (GSSX), and reduced glutathione (GSH) in coronary sinus plasma, immediately before cross-clamping the aorta (T0), and at 0 (T1), 15 (T2) and 30 (T3) min after unclamping it. In groups A and B a significant shift towards oxidation of redox status of glutathione (GSH/GSSX) at T1 vs T0 was observed. Glutathione redox ratio remained low in group B while in group A it returned to the basal value at T2 with a significant difference from group B at T2 and T3. No differences were observed between groups C and D. In conclusion, our data show that leukocyte-depleted reperfusion can afford a better myocardial protection in patients with left ventricular dysfunction, while it seems unnecessary in patients with normal EF. PMID- 8703493 TI - Multiple fistulas between coronary and pulmonary arteries. AB - An unusual case of a 62-year-old patient is presented with multiple coronary arteriovenous fistulas originating from both coronary arteries going into the pulmonary trunk. PMID- 8703494 TI - Isolated congenital left ventricular diverticulum. AB - Two cases of isolated left ventricular diverticulum are presented. A 12-year-old boy and a 7-year-old girl, both asymptomatic, were admitted to our Department for evaluation of an abnormal electrocardiogram and an abnormal cardiac silhouette, respectively. Both patients had the diagnosis confirmed by cardiac catheterization and angiography and underwent successful surgical correction of the abnormality. We discuss the diagnosis, prognosis, and surgical treatment of this entity and we advocate early surgical treatment for all diagnosed ventricular diverticula, even if asymptomatic. PMID- 8703495 TI - Rheumatic involvement of all four heart valves--preoperative echocardiographic diagnosis and successful surgical management. AB - Rheumatic disease of all four heart valves is rare. We report on a patient with quadrivalvar involvement by rheumatic disease, diagnosed preoperatively by echocardiography, who underwent successful surgical correction. The implications of organic involvement of the pulmonary valve by rheumatic process is discussed. PMID- 8703496 TI - Anomalous aortic origin of the right coronary artery. An unusual cause of angina pectoris and its surgical correction. AB - A 45-year-old female presented with typical recent-onset exertional angina pectoris. Subsequent investigation showed that the likely cause was an aberrant origin of the right coronary artery arising from the left coronary sinus. This anomaly is uncommon and is not usually associated with angina pectoris. Surgical rerouting of the origin of the right coronary artery produced complete resolution of ischaemia. PMID- 8703497 TI - Cerebral ischemia from thoracic goiter. PMID- 8703498 TI - Coronary artery bypass grafting in chronic renal dialysis patients: intensive perioperative dialysis and extensive usage of arterial grafts. PMID- 8703499 TI - [Predictive value of coronarography in the localization of arterial lesions responsible for future infarcts of the myocardium]. AB - OBJECTIVES: To evaluate the capability of coronary angiography, when performed in stable conditions, to predict which arterial segment will be responsible for future acute myocardial infarction. POPULATION: 17 patients (15 men and 2 woman), mean age 52 +/- 12 years, who suffered a myocardial infarction (AMI), and had been previously submitted to coronary angiography. After the myocardial infarction all the patients had a new angiography. METHODS: The coronary angiographies performed before and after the AMI were compared, by simultaneous visualization of the films, and the segment related to the AMI was determined with the help of ECG and ventriculography. Among the arterial segments with in the coronary angiography performed before the AMI and who were found to be related with it, three Groups were considered: A-with no angiographic lesion; B with less then 70% lesion; C-with a lesion equal or superior to 70% (angiographically significant). RESULTS: In nine patients the arterial segment related to AMI belonged to group A. In four patients it belonged to group B and in four to C. Segments with angiographically significant lesions were responsible for AMI in less then one fourth of the patients. CONCLUSIONS: Coronary angiography, when performed in stable conditions, has a low predictive value to determine the localization of the arterial segment related to future AMI. PMID- 8703500 TI - [Revascularization of the left coronary system with both internal mammary arteries in disease of the left main artery]. AB - Although the longterm patency of the internal mammary artery (IMA) has been clearly demonstred, some doubts have been cast as to the perioperative adequacy of its flow. Hence, the sole use of these conduits in patients with left main disease (LMD) has been cautioned. To clarify the significance of this problem in our own population, we have considered in this study, 110 patients with significant LMD subjected to isolated myocardial revascularization from November 1992 trough November 1994. These patients were retrospectively divided into two Groups based on the type of revascularization of the left coronary territory: Group I-35 patients (mean age 51.8 +/- 6.0 years) in whom both the left anterior artery and branches of the circunflex artery were grafted with the left and right IMAs, respectively; and Group II-85 patients (mean age 63.9 +/- 7.7 years) in whom the anterior descending artery received the left IMA and the circunflex system received saphenous vein grafts. No patient in Group I received venous grafts in the left coronary system and in four patients of Group II the rigth IMA was anastomosed to the right coronary artery. The operative mortality was 0% in Group I and 3.5% in Group II (p = NS). The rate of perioperative myocardial infarction was 2.9% and 2.3%, respectively. The incidence of other complications was also similar in the two groups with regards to the need for inotropes (8.6% in the Group I and 7.0% in group II), and the prevalence of arrhythmias (22.8% and 23.5%) but was marginally higher in Group I with regards to reoperation for haemorrhage (8.6% and 3.5%) and sternal dehiscence (5.7% and 2.3%). The mean time of hospital admission was 8.3 days for both groups. The use of both IMAs as the sole conduits for revascularization of the left coronary system in patients with LMD did not increased surgical risk. Also, the hypothesis of insufficient blood flow to the myocardium does not appear to be supported by this study. PMID- 8703501 TI - [Holter electrocardiography en pediatric cardiology: preliminary experience]. AB - PURPOSE: Evaluation of the diagnostic value of continuous electrocardiographic recording (Holter monitoring) in Paediatric Cardiology. STUDY DESIGN: Retrospective study based on the data records of the Arrhythmia Department regarding patients with less than 20 years of age with Holter monitoring performed between January 1983 and February 1995 (374 recordings). PATIENTS: 277 patients were studied, 163 were male and 114 female with a mean age of 12 +/- 5 years. Heart organic anomalies were present in 155 patients and 92 patients had previous cardiac surgery with cardiopulmonary bypass. Clinical indications for Holter recording were the evaluation of symptoms related to arrhythmias (38%), documented dysrhythmia (23%), arrhythmia detected on physical examination (18%) and evaluation of risk in patients without symptoms of arrhythmia (17%). METHODS: Holter recordings were performed during 24 hours with two channels (modified V1 and V5). RESULTS: Significant rhythm and conduction disturbances were found in 122 patients. These were ventricular premature complexes > or = 10/hour (27 patients), AV block (25 patients) and supraventricular premature complexes > or = 10/hour (22 patients). Holter recording revealed significant dysrhythmias in 52% of patients with previously documented dysrhythmia, in 43% of patients referred for assessment of risk without symptoms of arrhythmia, provided insight in 48% of requests due to arrhythmia detected on physical examination and 24% of requests for symptoms related to arrhythmias. In 13 patients the results of Holter recording led to therapeutic or diagnostic measures. CONCLUSIONS: Holter recording showed an overall sensitivity of 44% in the detection of significant arrhythmias and led to therapeutic and additional diagnostic measures in 11% of these patients. Holter recording seems to provide important information in the evaluation of arrhythmias in the paediatric age group. PMID- 8703502 TI - [Idiopathic ventricular fibrillation: apropos of 2 clinical cases]. AB - We describe two patients with idiopathic ventricular fibrillation recently evaluated to whom it was implanted a cardioversor desfibrillator system. It is our purpose to make a brief review of the etiology, diagnostic procedures, therapy and prognosis of this clinical entity, which is an important rare cause of sudden cardiac death. PMID- 8703503 TI - [Exertion syncopal crisis in the young, associated with idiopathic long QT syndrome]. AB - The cases of two young patients with repetitive syncopal attacks due to idiopathic long QT syndrome (ILQTS) are reported. Both had been primarily misdiagnosed for seizures. In one of the cases the QT interval in the ECG at rest was normal. The same patient had a previous episode of cardiac arrest with ventricular fibrillation. The treadmill stress test was of great value, revealing polymorphic ventricular tachycardia induced by exercise, and evaluating the efficacy of beta-blocking therapy in the follow-up. The ILQTS should be considered a possible etiology in any patient presenting with new onset seizures, especially in the young. The treatments were different in both cases. In the first one, the treatment with nadolol (100 mg od) revealed to be very effective with total remission of symptoms. The treadmill stress test performed 15 days after the beginning of treatment did not show any ventricular arrhythmias, and it was assumed that the patient was effectively protected against ventricular arrhythmias. After 4.5 months of follow up, no syncopal episodes occurred. In the second case due to young age, the frequency, and the severity of the attacks (cardiac arrest with ventricular fibrillation), and the inefficacy of beta adrenergic-blocking agents, the implantable cardioversor-defibrillator was the treatment chosen, although the beta blocking therapy was maintained to reduce the number of arrhythmic events. The ILQTS is a rare anomaly related to sudden cardiac death. The ILQTS is characterised by the association of several distinctive electrocardiographic features, among which prolongation of the QT interval is the best known. Life-threatening arrhythmia occurs usually under conditions of physical or psychological stress. Relatively effective therapies do exist and are represented by antiadrenergic interventions: beta-adrenergic blocking agents are the treatment of choice. When they fail, left sympathetic denervation or the automatic implantable cardioversor-defibrillator have also proved to be effective. PMID- 8703504 TI - [Renovascular arterial hypertension. From physiopathology to therapy]. AB - Understanding the pathophysiology, diagnosis, and management of renovascular hypertension (RVH) is of paramount importance due to the severity of hypertension (HT) and renal insufficiency (RI). Moreover, adequate treatment by surgery and/or endovascular intervention can improve HT and revert RI. The comprehension of the pathophysiology of RVH had its origin on the experiments of Goldblatt which led to the recognition of the renin dependent, volume dependent, and mixed types. A continuum seems to exist, from an acute phase, supported by the endocrine renin angiotensin aldosterone system, evolving towards a chronic phase sustained by the local renin angiotensin system. The involved vasoconstrictor and mitogenic mechanisms may contribute to the arterial remodeling. The most common forms of pathology, i.e. atherosclerosis, fibromuscular dysplasia (FD), and Takayasu's arteritis, and their natural history, are described. The prevalence of RVH, ranging from 0.2% to more than 25%, depending on the clinical situation, is evidenced. Clinical symptoms and signs and the most important diagnostic tests are pointed out: functional tests (captopril test, postcaptopril renography, scintigraphy, and renin determinations) and anatomical tests (intravenous digital angiography and intrarterial angiography). New imaging techniques are also referred. A diagnostic work-up based on the index of clinical suspicion is described. The therapeutic goal is the resolution of the two main problems of RVH: hypertension and ischemic nephropathy. Revascularization is becoming mandatory either by percutaneous transluminal angioplasty mostly for FD and atheromatous non-ostial stenoses, or by surgery, which is preferred for patients with ostial or peripheral stenoses, aneuryms, occlusions and concomitant aortic disease. A better knowledge of RVH allows, not only diagnosis and treatment of one of the most frequent types of secondary hypertension, but also the control of the resulting ischemic nephropathy. PMID- 8703505 TI - [Doppler echocardiography. Contribution for the analysis of its normal patterns]. PMID- 8703506 TI - [Reminiscing. The I centenary of the foundation of the Royal College of Surgery of Lisbon. 5-30 of December of 1925]. PMID- 8703507 TI - Entry of pathogens into the central nervous system. AB - The blood-brain barrier (BBB) is formed by the tight junctions of the cerebral capillary endothelium and the choroid plexus epithelium. The molecular anatomy of the tight junction resembles that of a polarized, transporting epithelium, suggesting some model cell culture systems can provide insight into traffic into the central nervous system. Pathogens target both the endothelium, causing encephalitis, and the choroid plexus, leading to meningitis. Routes of entry are diverse including paracellular and transcellular penetration. In addition, circulating microbial products can induce loss of BBB function. Understanding the heterogeneous molecular interactions between pathogens and the BBB may provide avenues to interrupt the devastating neurological sequelae that accompany central nervous system infections. PMID- 8703508 TI - Adaptation to life at micromolar nutrient levels: the regulation of Escherichia coli glucose transport by endoinduction and cAMP. AB - Free-living bacteria are expert in adapting to variations in nutrient availability, often using an array of transport systems of different affinities to scavenge for particular substrates (multiport). This review concentrates on the regulation of expression of different transporters contributing to multiport in response to varying nutrient levels. A novel mechanism of controlling bacterial transport affinity under sugar limitation is described. In particular, switching from glucose-rich to glucose-limited conditions results in Escherichia coli orchestrating outer membrane changes as well as the induction of a periplasmic binding protein-dependent (ABC-type) transport system. The changes leading to the high affinity transport pathway are directed towards uptake of rapidly utilisable concentrations and are optimal close to 10-6 M medium glucose. High affinity transport is absent under both glucose-rich 'feast' and glucose starved 'famine' conditions hence high affinity transporters are not simply repressed by excess nutrient. Rather, the improvement in glucose scavenging involves induction of genes in 2 distinct regulons (mgl/gal and mal/lamB) through synthesis of 2 different endogenous inducer molecules (galactose, maltotriose). Endoinducer levels are tightly controlled by extracellular glucose concentration at different glucose-limited growth rates. Aside from endoinducers, the elevated intracellular level of cAMP plays a role in induction of the high-affinity pathway but cAMP-mediated relief from catabolite repression is not itself sufficient for high affinity transport. In contrast to the repressive role of glucose when present at millimolar concentrations, micromolar glucose also leads to the induction of transport systems for other sugars, further broadening the scavenging potential of nutrient-limited bacteria for other substrates. PMID- 8703509 TI - Bacterial aminopeptidases: properties and functions. AB - Aminopeptidases are exopeptidases that selectively release N-terminal amino acid residues from polypeptides and proteins. Bacteria display several aminopeptidasic activities which may be localised in the cytoplasm, on membranes, associated with the cell envelope or secreted into the extracellular media. Studies on the bacterial aminopeptide system have been carried out over the past three decades and are significant in fundamental and biotechnological domains. At present, about one hundred bacterial aminopeptidases have been purified and biochemically studied. About forty genes encoding aminopeptidases have also been cloned and characterised. Recently, the three-dimensional structure of two aminopeptidases, the methionine aminopeptidase from Escherichia coli and the leucine aminopeptidase from Aeromonas proteolytica, have been elucidated by crystallographic studies. Most of the quoted studies demonstrate that bacterial aminopeptidases generally show Michaelis-Menten kinetics and can be placed into either of two categories based on their substrate specificity: broad or narrow. These enzymes can also be classified by another criterium based on their catalytic mechanism: metallo-, cysteine- and serine-aminopeptidases, the former type being predominant in bacteria. Aminopeptidases play a role in several important physiological processes. It is noteworthy that some of them take part in the catabolism of exogenously supplied peptides and are necessary for the final steps of protein turnover. In addition, they are involved in some specific functions, such as the cleavage of N-terminal methionine from newly synthesised peptide chains (methionine aminopeptidases), the stabilisation of multicopy ColE1 based plasmids (aminopeptidase A) and the pyroglutamyl aminopeptidase (Pcp) present in many bacteria and responsible for the cleavage of the N-terminal pyroglutamate. PMID- 8703510 TI - [Mitochondrial DNA polymorphism in the Han, Miao, Buyi, and Shui ethnic groups from Guizhou province]. AB - Sixteen restriction endonucleases were used to assay the mtDNA polymorphism in the Han, Miao, Buyi and Shui ethnic groups from Guizhou province. Thirty-one restriction patterns were observed in a total of 150 subjects studied, out of which HacII-13, EcoRV-3 and PstI-4 are first reported by this paper. From all restriction patterns disclosed by 16 enzymes, twenty-eight mtDNA types were obtained. UPG method and parsimony networks were employed to analyze the clustering relationships between types or populations. Following conclusions are obtained: the mt-DNA from the Shui nationality possesses the highest polymorphism; the Han and the Miao are most closely related; and the genetic relation between the Buyi and the Shui is somewhat distant. PMID- 8703511 TI - [Construction of a retroviral vector carrying human growth hormone gene and its introduction and expression in 3T3 and ES cells]. AB - In order to study the expression of growth hormone gene in heterologous cells, the retroviral vector pINS was used to construct the recombinant plasmid carrying the human growth hormone gene (hGH), and then it was introduced into mouse fibroblast cell line 3T3 and mouse embryonic stem cell line CCE. Levels of accumulation of hGH secreted into medium was quantified by a radioimmunoassay. Expression level of hGH gene varied considerably from one clone to another. Several 3T3 clones expressing hGH gene at high level were obtained, such as GHSNC 20, in which hGH concentration in medium was 3784ng/ml, Integration of exogenous gene was stable. Southern blot result showed that such kind of difference was not related to the number of integrated copies of the foreign gene. The expression of ES clones transfected with pINS-GH was undetectable, and integration of the foreign gene was unstable. Southern blot result showed that hGH gene really integrated into the cellular genome, and no rearrangement of foreign gene is found at present. PMID- 8703512 TI - [G gamma/A gamma, A gamma I/A gamma T ratios of fetal hemoglobin of Han, Hui, Uygur and Kazak newborns in Xinjiang and gene mapping analysis of two cases with abnormal ratio]. AB - The G gamma/A gamma, A gamma I/A gamma T ratios of fetal hemoglobin of 372 cord blood samples of newborns (Han 102, Hui 102, Uygur 99, and Kazak 69) in Urumchi were determined by HPLC. The means of G gamma ratios of 4 ethnic groups (Han, Hui, Uygur, and Kazak) were 66.83%, 68.33%, 70.44% and 69.70%, respectively. 4 cases (Hui 3 and Kazak 1) with low G gamma (< 50%), and 25 cases (Han 5, Hui 4, Uygur 11 and Kazak 5) with high G gamma (> 80%) were found. The occurrences of low G gamma in Hui and Kazak were 2.94% and 1.45% while in Han and Uygur no single case with low G gamma was found. Those of high G gamma in 4 ethnic groups were 4.9% (Han), 3.9% (Hui), 11.1% (Uygur) and 7.2% (Kazak), respectively. A gamma T heterozygotes in Han 21, Hui 23, Uygur 26 and Kazak 29 cases, and one A gamma T homozygote in each ethnic group were found. The average values of A gamma I were 56.83%, 55.58%, 50.94% and 54.68% respectively. The frequencies of A gamma T gene were 0.113, 0.123, 0.141 and 0.225, respectively. The gene arrangement of a low G gamma case was identified as-GA gamma I-/-G gamma-A gamma I-, and that of a high G gamma case as -G gamma-AG gamma-A gamma I(A gamma T)-/-G gamma-A gamma T(A gamma I)-. PMID- 8703513 TI - [The mitochondrial DNA diversity of pigs in southwest China]. AB - Twenty restriction endonucleases, ApaI, AvaI, BamHI, BclI, BglI, BglII, ClaI, EcoRI, EcoRV, HindIII, HpaI, KpnI, PstI, PvuII, SacI, SalI, ScaI, SmaI, XbaI, XhoI were used to investigate the mitochondrial DNA polymorphism of pigs from 11 localities in southwest China. Among the 28 animals analyzed, 27 restriction morphs were detected, which can be sorted into 6 haplotypes. The average genetic distance among haplotypes (P) is 0.004, and the value of average sequence variation in pig population (pi) is 0.122%. Our results suggest that the genetic diversity within the pigs is remarkably scarce. The pigs in southwest China may have origineted from a late common ancester, and were likely impacted by the founder effect. PMID- 8703514 TI - [Study on animal conservation-selection index]. AB - The concept of "conservation-selection index" was put forward based on the idea of Systematic Conservation that the conservation and selection must be combined in practice. Its calculation formula was also derived, which suits to various data and various aims of conservation and selection. PMID- 8703515 TI - [A cytogenetic study of four species of turtle from China]. AB - The karyotypes, C-banding and Ag-NORs of four species of turtle from China have been studied. The results demonstrated that Cuora aurocapitata has 2n = 52(14M + 2SM + 4ST + 6T / 26m), NF = 72, 8 + 5 + 13(karyotypic formulae). Its secondary constrictions (SC) located on No. 1 p inter of group I; C. trifasciata has 2n = 52(12M + 4SM + 4ST + 6T + 26m), NF = 72, 8 + 5 + 13; Cistoclemmys flavomarginatus has 2n = 52(16M + 4ST + 6T + 26m), NF = 72, 8 + 5 + 13. The SC is also on No. 1 p inter of group I; C. galbinifrons has 2n = 52(16M + 2SM + 4ST + 6T + 24m), NF = 74, 9 + 5 + 12. Four pairs of SC of C. galbinifrons are on No. 1 p inter, No. 3,7 p par and No. 6(X chromosome) q per of group I. The heteromorphic chromosomes related with sexuelity were discovered in C. galbinifrons, while there is no that in other three species. All the centrometric regions of the chromosomes for the four studied species showed the various degree staining of C-positive. Only one homologous pair of Ag-NOR, is found for C. galbinifrons, which locate on No. 5 q per of group II. The Ag-NOR, of other three species lies in No. 7 q ter of group I. The mechanism of the karyotypic evolution is discussed. Individual evolutional mechanism of an heteromorphic sex pair of chromosomes. PMID- 8703516 TI - [Amplified fragment length polymorphism of the VNTR locus COL2A1 in Chinese population]. AB - The amplifiable VNTR polymorphic system COL2A1 has been investigated in a Chinese Han population (n = 120) by the polymerase chain reaction (PCR) and PAGE horizontal electrophoresis followed by silver stain. In order to accurately identify COL2A1 alleles, a number of human allele ladders prepared by mixing DNAs extracted from different individuals of known COL2A1 genotypes were used. A total of 14 different alleles in 23 genotypes were observed in this Chinese Han population. Among them, four were new alleles disclosed in the present study. The results imply that COL2A1 locus may be served as a genetic marker in forensic haemogenetics as well as in anthropogenetics. PMID- 8703517 TI - [A survey on distribution of red cell blood group systems in naxi and primi ethnic groups]. AB - A survey of distribution of red cell blood group systems, including ABO, MNSs, Rh and P, was carried out on the Naxi and Primi ethnic groups in Yunnan province. The results based on 104 cases in each of the two ethnic groups showed that both Naxi and Primi possessed a high gene frequency r of 0.6082 and 0.6882, respectively, with gene frequency p = q. The gene frequency m of Naxi (0.8509) was found to be very high among the populations studied in China until now, only next to that of Lizu (0.8709). The most common phenotype of Rh system was CcDE- in both Naxi and Primi, with a quite high cDE frequency. No case of Rh negative was observed in the two ethnic groups. The P1 in Naxi approximated to that in Primi. The red cell blood group systems and their genetic distances suggested that the Naxi and Primi was genetically close to ethnic groups of North China, but different from those of South China. This fact suggests that these two ethnics groups originated from the North China. PMID- 8703518 TI - [Electrophoretic analysis of the protein patterns of the nuclear transplant rabbit embryos]. AB - The qualitative patterns of protein synthesis in nuclear transplant rabbit embryos were examined by SDS-polyacrylamide gel electrophoresis followed by silver staining. The results indicated that the qualitative pattern of several protein synthesis in NT embryos was very different from the protein pattern of donor morulae or recipient oocytes, and also not as the same as the protein pattern obtained from fertilized ova at pronuclear formation stage. After fertilization or nuclear transfer, several maternal proteins were no longer synthesized or decreased in the embryo, and some new bands were observed and several protein synthesis were increased obviously. The most intriguing aspect of this study was the observation that all major changes in the protein pattern took place after fertilization or nuclear transfer and were rather similar. It is suggested that the gene activities of the donor nucleus from rabbit morulae are reprogrammed by the oocyte cytoplasm in current nuclear transfer technology, but the reprogramming is incomplete. This paper stresses on the paternal effect during fertilization on gene expression in nuclear transplant rabbit embryos. PMID- 8703519 TI - [Molecular cloning of cellulase gene from the bacillus]. AB - The CMCase gene from the Bacillus sp. strain E2 was cloned into E. coli DH5 alpha F' using plasmid pBR322 as a vector. Recombinant plasmid pBG3 was isolated from transformants producing CMCase, and the cloned CMCase gene was found to be in a 4.0kb HindIII fragment. The CMCase gene was subcloned in a functional state on a 2.4kb DNA fragment. The optimum pH and temperature for activity of CMCase encoded by pBG3 were pH 6.5 and 55 degrees C respectively. The CMCase could not hydrolyze any natural cellulose examined. Eighty-three percent of the CMCase was detected in the inner cellular fraction, 9% and 8% were in the periplasmic space and medium respectively. Result of Southern hybridization indicates that the cloned DNA fragment was derived from Bacillus sp. strain E2 chromosomal DNA. PMID- 8703520 TI - Detecting and managing depression in older people. PMID- 8703521 TI - Corticosteroid injections for lateral epicondylitis: a systematic overview. AB - BACKGROUND: Lateral epicondylitis (tennis elbow) is a common complaint, for which corticosteroid injections are a frequently applied therapy. However, there were no up-to-date reviews available that systematically addressed the effectiveness and adverse effects, including questions concerning optimal timing of injections and composition of the injection fluid. AIM: The aim of the study was to assess the effectiveness of corticosteroid injections in the treatment of lateral epicondylitis (tennis elbow) by systematic review of the available randomized clinical trials. DATA SOURCES: The data sources used were randomized clinical trials identified by literature searches of the MedLine (1966-1994) and Embase (Exerpta Medica) (1980-1994) databases for the keywords epicondylitis, tendinitis and elbow, injection. References given in relevant publications were further examined. STUDY SELECTION: The criteria for selecting studies were as follows: randomized clinical trials (treatment allocation in random or alternate order); one of the treatments to include one or more corticosteroid injections (additional interventions were allowed); participants suffering from lateral epicondylitis; and publication in English, German or Dutch. Abstracts and unpublished studies were not included. DATA SYNTHESIS: Methodological quality was assessed by means of a standardized criteria list (range 1-100 points). The extracted outcomes were the general conclusion drawn by the authors of the reports on the trials, and the success rates at the various follow-up points as (re)calculated by us. The success rates were subsequently graphically displayed and statistically pooled. Separate stratified analyses were conducted according to a predetermined analysis plan. RESULTS: Twelve randomized clinical trials were identified. The median methodological score was 40 points, indicating an overall poor to moderate quality. The pooled analysis indicated short-term effectiveness (2-6 weeks): pooled odds ratio (OR) = 0.15 [95% confidence interval (CI) 0.10 0.23], chi 2 [degrees of freedom (df = 5) = 13.3], indicating statistical heterogeneity. At longer term follow-up, no difference could be detected. The studies of better methodological quality indicated more favourable results than those of lesser methodological quality. The most suitable corticosteroid to use as well as dosage, injection interval and injection volume could not be derived from the various trials. CONCLUSION: The existing evidence on corticosteroid injections for the treatment of tennis elbow is not conclusive. Many trials were conducted in a secondary care setting and clearly had serious methodological flaws, and there was statistical heterogeneity among the trials. Corticosteroid injections appear to be relatively safe and seem to be effective in the short term (2-6 weeks). Although the treatment seems to be suitable for application in general practice, further trials in this setting are needed. As yet, questions regarding the optimal timing, dosage, injection technique and injection volume remain unanswered. PMID- 8703522 TI - The capture of socioeconomic data in general practice. AB - BACKGROUND: It is common practice to record the reasons why patients have an encounter with the practice, but the collection of socioeconomic data with which to link this morbidity data is less easy to achieve. AIM: To describe the social enquiry used in the Fourth National General Practice-based morbidity study (1991 1992) and to consider its effectiveness for use in practice. METHOD: Socioeconomic data were collected suing a structured questionnaire administered by a trained interviewer. Data were provided by both consulting and non consulting registered patients. RESULTS: The interview technique proved to be acceptable to patients, interviewers and general practitioners, simple to administer, and inexpensive to collect. Eighty-three per cent of the 502,000 people included in the study provided social and occupational data. Less than 1.5% of patients refused to be interviewed. Fifty-four of the 60 practices achieved the target level of 90% of registered patients being successfully interviewed. CONCLUSION: A method of socioeconomic data collection based on that used in the 1991-1992 study would be of benefit for health care planning, allocation of resources, design of performance indicators and epidemiological research. PMID- 8703523 TI - Links between mental health care professionals and general practices in England and Wales: the impact of GP fundholding. AB - BACKGROUND: Fundholding general practitioners are able to determine the type of contracts they place with providers of mental health care, and are able to employ some categories of mental health care professionals directly. The impact of this on the care of the mental health of patients in non-fundholding practices is not yet fully known. AIM: A survey was undertaken of 100 fundholding general practices and 100 similarly sized non-fundholding practices in order to investigate the changes in mental health provision made by general practitioners. METHODS: A sample of 100 fundholding general practices in England and Wales was randomly chosen from the list supplied by the Association of Fundholders and matched to a similarly randomly chosen sample of non-fundholding practices. Postal questionnaires were sent to the senior partner and to the practice manager in each practice. RESULTS: The number of mental health care professionals who are either employed by or attached to general practices, or who visit the general practice on a regular basis appears to have increased substantially since 1991. This increase was particularly marked in fundholding practices. The results suggest that general practitioners with specific links to particular mental health care providers were more satisfied with the service provided by the mental health care team, and more likely to increase referrals to that service in the last 2 years, than general practitioners without such links. There was little evidence to suggest that increasing the number of mental health care professionals in primary care had brought about a major reduction in referrals to psychiatrists. CONCLUSION: General practitioners, particularly fundholders, are increasing their links with mental health professionals, and community psychiatric nurses, psychiatrists, psychologists and counsellors are spending more time either based in general practice or visiting regularly. While the shift of resources to primary care, particularly to fundholders, may increase the treatment options available to patients with less severe illnesses, this may have the effect of reducing the services available for the long-term and severely mentally ill. PMID- 8703524 TI - Pilot study of the acceptability of cystic fibrosis carrier testing during routine antenatal consultations in general practice. AB - BACKGROUND: In 1989, the gene for cystic fibrosis was cloned and it became possible to detect carriers of the gene among the general population, including pregnant women. AIM: The aim of the pilot study was to assess the acceptability of integrating cystic fibrosis carrier testing into antenatal care by general practitioners at the first booking appointment. METHOD: Between 1 September 1991 and 31 August 1992, inclusive, all patients receiving routine antenatal care in a two-partner training practice in south Manchester were offered carrier testing for cystic fibrosis using a computer protocol for antenatal care developed by the practice. A questionnaire including a Spielberger state-trait anxiety inventory was sent to patients 2 weeks after they received the results of their carrier test, and interviews with the patients in their home were carried out 4 weeks and one year after they received the result. RESULTS: All but one patient (75 out of 76) booking before 14 weeks of pregnancy accepted the offer of cystic fibrosis carrier testing, and 96% (72 out of 75) felt that they had made the right decision and that they had enough time for discussion with their general practitioner before testing. CONCLUSIONS: Cystic fibrosis carrier testing can be successfully integrated into the antenatal booking appointment in general practice and is acceptable to patients. This is a model for other genetic screening opportunities resulting from advances in medical genetics. PMID- 8703525 TI - Family systems ideas in the 10-minute consultation: using a reflecting partner or observing team in a surgery. AB - BACKGROUND: Family therapy and the ideas that underlie it have not had much impact on general practice, although there is good reason to think this could be a useful approach. AIM: As a group of general practitioners (and a practice nurse) with experience of family therapy, we were interested in demonstrating whether family therapy methods could usefully inform general practice consultations. METHOD: Two surgeries were observed by the general practitioner's colleagues. RESULTS: Selected cases are discussed to illustrate the impact of the use of family therapy ideas and a reflecting colleague or team. CONCLUSIONS: Family therapy ideas can be a helpful framework within which to think about both demanding and ordinary consultations. Inviting a colleague to contribute respectfully could be useful both as a method of teaching, and in resolving difficult or faltering consultations. PMID- 8703526 TI - Community care of patients with schizophrenia: the role of the primary health care team. AB - Schizophrenia is a severe, chronic mental disorder that usually begins in early adulthood. Recurrent relapse leading to long-term psychological and social disability means that patients may require intensive community support. Despite a recent fall in the overall numbers of patients consulting their general practitioner with mental disorders, presentations by those suffering from severe mental disorders have risen. This review encompasses the role of general practitioner in the management of schizophrenia, considering in turn drug and psychological therapies, family interventions, innovations in care, the effects of community care developments, and the liasion between primary health care and mental health professionals. There is a need for further research in the area of family-practice-based interventions involving general practitioners and the practice team. PMID- 8703527 TI - Domestic violence: a hidden problem for general practice. AB - Domestic violence is a common problem that may affect more than a quarter of women. It is a complex area in which to undertake research. Studies often focus on selected populations and exhibit a diversity of design, making comparison difficult. This review focuses on physical violence by men against women partners or ex-partners, and exemplifies important issues for general practitioners. Domestic violence frequently goes undetected. This may be the result of doctor's fears of exploring an area perceived as time-consuming, where knowledge is lacking and where they feel powerless to 'fix' the situation. Women may not reveal that they are experiencing violence, sometimes because doctors are unsympathetic or hostile. Nevertheless, women wish to be asked routinely about physical abuse and want to receive immediate advice and information about their options if necessary. Women experience a range of health and social problems in association with domestic violence, including depression, anxiety, substance abuse and pregnancy complications. However, none of these features is specific enough to be useful as an indicator of violence. Therefore, doctors should routinely ask all women direct questions about abuse. This recommendation can be incorporated into guidelines, which should be implemented widely in the UK, to improve the care of women experiencing domestic violence. In parallel with this, the educational needs of general practitioners should be addressed. Further research is needed to establish the prevalence of domestic violence in women presenting to general practice and to investigate how the problem is currently being addressed. If progress is to be made in tackling domestic violence, action within primary care is just one part of this: a fundamental change in the attitudes of men towards women is required. PMID- 8703528 TI - Should general practitioners have any role in maternity care in the future? AB - Maternity services in England are currently being reorganized. The success of the changes will be judged against the recommendations of the Changing Childbirth report. This paper describes the nature of maternity care and of general practice. It is argued that maternity care provision by general practitioners is a central and essential part of British general practice. Specifically, it is shown how general practitioners can help to achieve the objectives of the report, and thus, have a future role. It is suggested that all general practitioners who wish maternity care to remain an essential part of general practice need to argue the case with providers and purchasers. If they do not, then it is quite likely that general practitioners will be increasingly excluded as the commissioning and contracting mechanisms become more effective with midwives providing low-risk care and consultant obstetricians high-risk care. PMID- 8703529 TI - Medicine--a healing or a dying art? AB - That medicine is a science is the popular belief, and this has been reinforced by the advent of 'evidence-based medicine', which assumes that scientific proof is required to support medical practice. However, the view of science implied is a narrow one, foreign both to pure scientists and to artists, and the art of medicine is devalued by the approach. There are both important differences and important similarities between science and the arts. The arts should contribute to evidence-based practice and education along with science, and have a role in many aspects of medical practice. PMID- 8703530 TI - On throwing out the baby and keeping the bathwater. PMID- 8703531 TI - Multimedia teaching and learning: transforming the culture of higher professional training? PMID- 8703532 TI - Periconceptual folate supplementation. PMID- 8703533 TI - Crisis in recruitment. PMID- 8703534 TI - Aspirin in acute myocardial infarction. PMID- 8703535 TI - Chlamydial infection in women. PMID- 8703536 TI - Cold chain storage. PMID- 8703537 TI - Effect of leaflets on contraception. PMID- 8703538 TI - [Time factor and curietherapy: the example of epidermoid carcinoma of the mobile tongue]. AB - Experimental evidence exists of a dose-rate effect relationship in low dose continuous irradiation of cell population. In 1952, Paterson drew from his clinical experience on radium 226 implant, a dose-time iso-effect curve. Several authors, reviewing series of squamous cell carcinomas of the mobile tongue, implanted by radium 226 or iridium 192, did not confirm Paterson's hypothesis. Recently, a retrospective analysis of T1 and T2 squamous cell carcinomas, all treated exclusively by iridium 192 at the Henri-Mondor Hospital (Creteil, France), suggested a possible dose-rate effect relationship below 1 Gy/h. However, the role of dose-rate remains unclear. Further experience is needed before definitive conclusions can be drawn. PMID- 8703539 TI - [Cancer of the anal canal]. PMID- 8703541 TI - [The role of radiotherapy in the treatment of rectal cancer]. PMID- 8703540 TI - [Arterial stenosis after radiotherapy]. AB - Published reports of arterial stenosis following radiotherapy are reviewed. In oncological practice, this complication is rare but experimental studies have demonstrated the role of irradiation in producing arterial lesions. The histological specificity and the mechanisms of radiation and related arterial lesions are discussed. Atherosclerosis risk factors and chemotherapy could have a synergic role on artery stenosis. Cases reported of arterial stenosis after radiotherapy include subclavicular artery after breast cancer, carotid artery after head and neck cancer, coronary artery and abdominal aorta or its trunks after pelvic and abdominal irradiation. The radiotherapy parameters described are not unusual. Therapeutical modalities and their indications are presented. PMID- 8703542 TI - [Association of 5-FU, CDDP and hypofractionated radiotherapy in recurrences of subtentorial astrocytomas and malignant gliomas in adults]. AB - Twenty-seven patients who had a relapse of astrocytomas or supra-tentorial malignant gliomas, previously treated by radio-surgical combination or exclusive irradiation, underwent a combination of 3 courses of 5-FU, cis-platyl and hypofractionated irradiation. No objective response was observed, but there was a progression of the tumor in 33% of the cases, and no change in 67%. Overall median survival was 7 +/- 1.5 months. The 20 patients with grade III or IV astrocytomas had a lower median rate than the 7 patients with grade II astrocytomas or oligodendrogliomas: 6 +/- 1.8 months versus 18 +/- 3.2 (P = 0.16). Median survival was 14 +/- 2.7 months for neurological responders and 6 +/ 2.1 months for non-responders (P = 0.017). PMID- 8703543 TI - [Malignant non-Hodgkin's lymphoma of the conjunctiva. Apropos of 14 cases treated at the Institut Gustave-Roussy]. AB - Non-Hodgkin lymphomas (NHL) of the conjunctiva are rare. Radiotherapy is usually considered to be the choice of treatment in such situations. Reported here are 14 cases of conjunctival NHL, treated at the Institut Gustave-Roussy from 1981 to 1988. Mean age of the patients was 53 years. The series comprised 7 stages of IE, 4 stages of IIE and 3 stages of III. Histological types were mostly "low-grade" according to the Working Formulation (9 cases out of 14). The irradiation technique took advantage of a customized facial lead mask and of a direct anterior electron beam. A small cylindrical lead block, hanging some millimetres above the eye, shielded the cornea and the lens. Local control was achieved for 17 eyes treated (out of 19 irradiated), with a follow-up ranging from 5 to 90 months. The 2 relapses occurred in the same patient, and could be treated by a second line irradiation of the same type. Immediate tolerance was good. Late sequelae were rare and actually included 4 cases of cataract which clearly could not be entirely attributable to the treatment. In conclusion, this sophisticated technique appears to be efficient and non-toxic, since it was able to achieve an almost 100% local control rate, together with a very low percentage of late complications. PMID- 8703544 TI - [Radiotherapy of inoperable esophageal cancer. Retrospective analysis apropos of 108 cases]. AB - Between 1976 and 1986, we treated with curative intent, 108 patients with inoperable esophageal cancer. Eighty-three patients (77%) were treated with radiation therapy alone with a mean total dose of 61 Gy and 25 (23%) patients were treated with induction chemotherapy followed by radiotherapy. Actuarial survival, site of failure, sequellae of treatment, and prognostic factors were analysed in this retrospective study. Survival rates at 2, 3 and 5 years were 23, 16 and 9%, respectively. Local failure occurred in more than 61% of documented failures. Patients with tumor located in the upper third of the esophagus, or with total tumor stenosis or poor performance status had low results, but the difference was not statistically significant. Patients treated according to CT scan plans had a 43% 3-year survival rate; this difference is statistically significant (P = 0.0001). Induction chemotherapy improved one year local control and survival rate (60%), but no long term survival benefit was observed. We conclude that inoperable esophageal cancer can be treated successfully by radiotherapy with a curative intent. We also discuss the role that CT scan planning can play in the effective treatment of these patients. PMID- 8703545 TI - [Postoperative radiotherapy in soft tissue sarcomas of the limbs. Analysis of irradiation volumes and doses in a series of 31 cases]. AB - From October 1985 to October 1988, 31 patients with a soft tissue sarcoma of extremities were treated in our institute by conservative resection and post operative radiation therapy. The initial target volume encompasses the entire musculoskeletal area occupied by the tumor. The compartmental definition of this initial target volume is based on clinical, radiographic, histopathologic and operative findings. We analyzed the irradiated volume in terms of the anatomical definition and feasibility. We then evaluated the distribution of the dose in the irradiated volume. The variation of the dose is always less than 10%. Accordingly, the range of dose in the irradiated volume varies from 45 to 50 Gy in the compartmental volume after a complete resection, and from 55 to 60 Gy in the tumor bed after a marginal or incomplete resection. For this series, the median follow-up is 27 months. We observed 3 distant metastases and 1 progressive disease in the irradiated volume. No locoregional recurrence appeared in the margins or outside the irradiated volume. These preliminary results validate the compartmental definition of the initial target volume. Furthermore, they point out the lack of opportunity to encompass tendinous insertions in the irradiated volume and to boost the scar. Finally, the definition of optimal dose for the control of microscopic residual tumor is discussed. PMID- 8703546 TI - [Treatment of cervical lymph nodes in lip cancers]. AB - From 1970 to 1985, 299 patients with lip cancer were examined, treated and followed-up at the Bergonie Foundation. In most cases, interstitial radiotherapy was used and the local control rate reached 94%. Local recurrences could usually be treated again, so that the final local control rate was 99%. However, the management of neck nodes remains controversial in some cases: 5.6% only of T1-2 N0 stages developed neck nodes, which were successfully controlled in 78.9% of cases and it was therefore considered that the preferred option should be to maintain the patients under close follow-up. For T3 N0 cases, of which 17.6% would evolve, it was considered that a sub-mental and sub-maxillary neck node dissection was advisable when performance status was satisfactory and when a close follow-up was difficult. The recurrence rate was 40% for patients with palpable neck nodes who were often submitted to a combined radio-surgical treatment. In such cases, recurrences were controlled in 22% of patients who died 7 times out of nine. PMID- 8703548 TI - [Biological equivalency of high single doses used in intraoperative irradiation]. AB - Intra-operative radiotherapy is being used more and more frequently; this raises the problem of the equivalence of these large doses delivered in a single fraction, especially when a second line, complementary external irradiation is planned afterwards. The linear quadratic (LQ) model is probably the most convenient way to compare 2 irradiation schedules delivered with different doses per fraction. However, in the case of intra-operative radiotherapy, one should question the use of the LQ equation; this model actually predicts a continuous bending of the survival curve, while most experimental curves show a trend towards exponential at high dose levels. As a result, with the very large doses (20-30 Gy) given here, the LQ model would lead to overestimate the efficacy--and the toxicity--of intra-operative irradiation. We thus propose to calculate the equivalent fractionated doses by combining 2 models; the LQ model, considered to be reliable up to 7 Gy, and the 2 components' target model, which tends towards an exponential at high dose level. Moreover, we must take into account the hypoxic component of the tumour, since the single fraction, intra-operative technique cannot benefit from the reoxygenation occurring during conventional fractionated irradiations. Calculations indicate that for a partially hypoxic tumour, the effect of a large dose (20-30 Gy) is very similar to the one which would be achieved by the same dose given in a conventionally fractionated way, increased by the dose necessary to compensate for the proliferation occurring during treatment. For healthy tissues, assuming that all cells are well oxygenated, the equivalent fractionated dose is, in particular, much higher for late responding tissues. IN CONCLUSION: 1) there is little hope for sterilizing any carcinoma with only one fraction of 20-30 Gy; 2) however, intra-operative irradiation, focussing on a small volume, could contribute to the tumour eradication; 3) the intra-operative irradiation technique should, as much as possible, spare the neighbouring normal tissues, regarding the risks of long term complications. PMID- 8703547 TI - [Curietherapy of the Eustachian tube using Ir-192 in chronic otitis of tubal origin]. AB - Failure of chronic otitis surgery, especially tympanoplasty, are mainly related to Eustachian tube dysfunction. A new technique of anti-inflammatory irradiation of the Eustachian tube was designed in 1985 in Creteil to improve the Eustachian tube function. The Eustachian tube is catheterized during the tympanoplasty using a plastic tube, 1.6 mm in diameter; this tube is closed at the external end. The other end of the tube is open and goes through the opposite nasal cavity. The plastic tube is loaded 24 h later with a 4.5 cm long irridium 192 wire to deliver a dose of 3 Gy on the reference isodose, 4 mm in diameter, in approximately 3 h. The tube is then gently withdrawn through the nasal cavity. Seventy-four patients, 16 years old or more, took part in the study. The method was performed successfully in 62 patients. Fifty-six patients were followed up; mean follow-up was more than 20 months. The follow-up assessment included otoscopy, audiometry, impedancemetry and residual Rinne measurement. Otoscopy results were adequate in 86% of patients. Hearing was improved in 44% of patients. Impedancemetry was adequate in 70% of patients and residual Rinne lower than 20 dB in 56%. These results are better if compared with those of a series of 30 tympanoplasties performed without intracavitary irradiation in 1984: the tympanoplasty failure rate then was 73%. In conclusion, this new procedure proved to be safe and simple and led to an improvement of the functional results of complex tympanoplasty. PMID- 8703549 TI - [Present status of radiotherapy]. PMID- 8703550 TI - [French Society of Oncologic Radiotherapy]. PMID- 8703551 TI - [Predictive tests of radiocurability: towards a custom-made radiotherapy?]. AB - Up to now, radiation oncologists had at their disposal only a number of well known histological and clinical factors in order to define the optimal dose which should be delivered to a given tumor. Recently, radibiological studies have suggested additional parameters which may play a major role in tumor radiocurability. These parameters are: the number of clonogenic cells, intrinsic radiosensitivity, hypoxia and proliferation kinetics. "Predictive" tests are being developed and evaluated for each of these parameters. The more advanced studies deal with intrinsic radiosensitivity; preliminary data show impressive variations in radiosensitivity within groups of clinically "homogeneous" tumors. Should these tests prove to be reliably predictive of radiocurability, it will be possible, in the near future, to propose to any given patient a "custom-made radiotherapy" adapted to the precise features of his or her tumor. PMID- 8703552 TI - [Cisplatin and radiotherapy: towards a new therapeutic system?]. PMID- 8703553 TI - [Corticobasal degeneration: clinical features]. PMID- 8703554 TI - [Pathophysiology underlying apraxia in corticobasal degeneration]. PMID- 8703555 TI - [Neuropathology of corticobasal degeneration]. PMID- 8703556 TI - [Topographic organization of the ventral striatum afferent projection from amygdaloid complex and hippocampal formation]. AB - The organization of the striatal afferent fibers from the amygdaloid complex and hippocampal formation was studied in the monkey with particular emphasis on specific projections of the ventral striatum. Retrograde tracers were injected into the different regions of the ventral (limbic) striatum and dorsolateral (sensorimotor) striatum. Labeled neurons were observed in the various regions of the amygdaloid complex and hippocampal formation. The medial ventral striatum received dense projections from the amygdala (the basal nucleus and the magnocellular division of the accessory basal nucleus), and the hippocampus (subiculum, CA1 and CA3). The shell of the nucleus accumbens (calbindin-D28k negative region) also received dense projections from the amygdala (the basal nucleus and the magnocellular division of the accessory basal nucleus), and the hippocampus (subiculum). The injections into the core of the nucleus accumbens showed scattered labeled neurons in the amygdala, and only a few labeled neurons in the hippocampus. The lateral ventral striatum received few inputs from the amygdala and hippocampus. In contrast to the ventral striatum, the dorsolateral striatum received no projection from the amygdala or the hippocampus. The connectional similarities between the medial ventral striatum and the shell of the nucleus accumbens suggest that although the medial ventral striatum is not calbindin-D28k negative, it may be regarded as a transitional zone between the shell and the rest of the ventral striatum. PMID- 8703557 TI - [The relationship between cell proliferation activity and secretory activity in pituitary adenoma--a review of 63 cases]. AB - Determination of the cell proliferation activity of neoplasm is useful in making a prognosis. Immunohistochemical detection using MIB-1 monoclonal antibody has recently allowed us to assess tumor cell proliferation easily, because it can be performed on paraffin-embedded specimens and the results have been demonstrated to be positively correlated with the results of PCNA staining. In this study, surgical specimens of 63 pituitary adenomas were examined by immunohistochemical staining with MIB-1 monoclonal antibody. Twenty-nine cases were non-functioning pituitary adenomas, 20 were prolactin (PRL)-producing pituitary adenomas, and 14 were growth hormone (GH)-producing pituitary adenomas. The MIB-1 positive rates of the pituitary adenomas ranged from 0% to 6.46%. In the non-functioning pituitary adenomas, the MIB-1 positive rates ranged from 0% to 4.55% (mean : 0.76%), in the PRL-producing pituitary adenomas the MIB-1 positive rates ranged 0% to 6.46% (mean : 0.91%), and in the GH-producing pituitary adenomas the MIB-1 positive rates ranged 0% to 1.28% (mean: 0.58%). There were no significant differences between these values according to the results of the Wilcoxon signed rank test. Although the size of the non-functioning pituitary adenomas was not correlated with their MIB-1 positive rate, tumor size was closely correlated with the interval between the onset of the initial symptoms and the date of surgery. In the PRL-producing pituitary adenomas, the MIB-1 positive rate was not correlated with serum PRL levels as an index of secretory activity, but was correlated with the PRL staining positive rate. Preoperative bromocriptine therapy proved effective in reducing tumor size and serum PRL levels, but had no effect on the MIB-1 positive rate. In the GH-producing pituitary adenomas, the MIB-1 positive rate was not correlated with serum GH levels as an index of secretory activity, but was closely correlated with the GH staining positive rate. All three groups included both invasive and noninvasive tumor types, but there were no close statistical correlations between the three tumor types. PMID- 8703558 TI - [Electrophysiological evaluation of the demented state and the nootropic effect of TA-0910 by "automated fluctuation analysis" of the high-frequency EEG- relation between neuropsychological test results and Lorentzian parameters]. AB - This study was performed to evaluate the demented state and the nootropic effect of TA-0910 by "Automated Fluctuation Analysis" of the human high-frequency EEG. Correlations between Lorentzian parameters S1 and S2 and the results of neuropsychological tests was investigated. The subjects of this study were 12 demented patients, four males and eight females, aged 57 to 80 years. After obtaining their informed consent a daily dose of 10 mg or 20 mg of TA-0910 was assigned to these subjects one by one. The Kana Hiroi test for pre-frontal lobe function and the Mini Mental State (MMS) test for function of the posterior part of brain were administered before and after the medication period. Particular attention was paid to the consciousness level of the subjects during EEG recording, and EEGs were stored on magnet tape. Statistical analysis was performed by Student's t-test. Improvement in the MMS score from 16.4 +/- 6.2 (mean +/- S.D.) before treatment with TA-0910 to 24.3 +/- 5.7 after treatment was significant (p < 0.001), and a tendency toward dose-dependency was noted. The changes in the traditional EEG did not coincide with these findings but Lorentzian parameters S1 and S2 and the results of the neuropsychological tests correlated well. All of the subjects failed the Kana Hiroi test both before and after the medication period, and in keeping with this finding, the normal frontal dominant pattern of the S1 topographical display was not detected. Furthermore, not only was the elevation of the S2 value significant but the correlation with the MMS scores was also significant, especially in the posterior and right half of the brain. These findings strongly suggest that 1) "Automated Fluctuation Analysis" is very useful not only for evaluating the demented state but for evaluating nootropics as well and 2) TA-0910 activation of subcortical structures, especially A-10, appears to be the mechanism to action. PMID- 8703559 TI - [An unusual autopsy case of corticobasal degeneration--with special reference to clinicopathological differentiation from progressive supranuclear palsy and slowly progressive aphasia]. AB - We report an unusual autopsy case of corticobasal degeneration. The patient was male who was 67 years old at the time of his death. He developed clumsiness of his right hand at age 65. Neurological examination at age 66 revealed supranuclear ophthalmoplegia of upward gaze and parkinsonism. Progressive supranuclear palsy was suspected. The patient's clinical course was rapidly progressive. Dementia developed nine months after the onset of the disease, and the patient manifested apallic syndrome at 21 months and died of pneumonia at about 25 months. At autopsy the brain weighed 1370g. Macroscopic examination revealed prominent atrophy of the anterior and parietal lobes, particularly of the pars opercularis in the left inferior frontal gyrus, and there was marked depigmentation of the substantia nigra. Histological examination showed neuronal loss, glial proliferation, spongy state, and ballooned neurons in the frontal and parietal lobes. Neuronal loss with glial proliferation was conspicuous in the striatum, pallidum, thalamus, and substantia nigra. There was slight neuronal loss in the dentate nucleus and locus ceruleus. Staining by Gallyas-Braak method revealed massive appearance of argyrophilic threads. This case is important in terms of the clinical differentiation from progressive supranuclear palsy, and is also interesting in regard to the pathological correlation with slowly progressive aphasia. PMID- 8703560 TI - [Hydroxyl radical generation and post-ischemic reperfusion brain damage implications for neuroprotection]. AB - We report the effects of the free radical scavengers, salicylic acid and bifemerane HC1, on the survival of hippocampal neurons in the Mongolin gerbil model of ischemia-reperfusion brain damage. In addition to performing histological examinations, we used the salicylate-trapping method to measure the amounts of hydroxyl radicals generated in the cerebral cortex and the hippocampus. Ischemia-reperfusion significantly reduced the number of neurons in the CA1 region of the hippocampus. The decrease was largely prevented by pretreating the animals with either salicylate, 20 mg/kg, or bifemerane HC1, 25 mg/kg. Administering salicylate to the animals 30 minutes after reperfusion was also effective, but bifemerane HC1 was ineffective when given after reperfusion. Two minutes after post-ischemic reperfusion, 2,3-dihydroxybenzoic acid levels were significantly elevated in the cerebral cortex and the hippocampus, and 2, 5 dihydroxybenzoic acid levels were significantly elevated in the hippocampus. The increase in 2,5-dyhydroxybenzoic acid in the hippocampus was suppressed by bifemerane HC1 given 30 minutes before exposure to ischemia. These results suggest that hydroxyl radicals are generated in the gerbil model of ischemia reperfusion brain injury, and that salicylate and bifemerane HC1 partially prevent the loss of neurons in the hippocampus. Hydroxyl radical scavengers may be useful in reducing neuronal damage associated with ischemia-reperfusion injury. PMID- 8703561 TI - [A case of predominant right thalamic infarction with severe verbal memory disturbance]. AB - We encountered a 45-year-old right-handed man who had suffered a predominant right thalamic infarction and complained of memory loss. Performance on the Miyake Test (recall of ten pairs of related and unrelated words), the Rey Osterrieth Complex Figures, the Benton Test of Visual Retention and the Wechsler Memory Scale-R disclosed a severe verbal memory disturbance associated with a little, if any, visual memory disturbance. An MRI study revealed bilateral lesions limited to the thalamus involving most of the right anterior nucleus (AN), mediodorsal nucleus (MD), ventrolateral nucleus (VL), and centromedial nucleus (CM), as well as a small part of the left MD, and CM. HM-PAO-SPECT scans showed areas of decreased cerebral blood flow not only in the right thalamus but in the medial and basilar region of the right temporal lobe. It is noteworthy that our patient had a predominant right thalamic lesion and exhibited a severe verbal memory disturbance rather than visual memory disturbance. This suggests that the right hemisphere is dominant for verbal memory function in this patient. PMID- 8703562 TI - [Favorable prognosis in a case of recurrent cerebral infarctions caused by chronic disseminated intravascular coagulation due to ovarian carcinoma]. AB - We report a 53-year-old woman who developed a cerebral infarction in the left middle cerebral artery area. Although she did not have a hemorrhagic tendency, the results of her examination showed a chronic disseminated intravascular coagulation (DIC) and a left ovarian carcinoma. Gabexate mesilate (FOY) and nafamostat mesilate (FUT) were not effective for her DIC. One month after admission, she had another cerebral infarction in the right posterior cerebral artery area. She was treated by heparin sodium and her chronic DIC improved. Then, she was operated on for ovarian carcinoma; the histologic finding was clear cell adenocarcinoma. No recurrence of DIC nor cerebral infarction was observed postoperatively. Patients with cerebral infarctions caused by chronic DIC due to ovarian carcinoma have been reported in the literature, but few patients had a favorable prognosis such as the present case. Heparin therapy appears to be the treatment of choice for chronic DIC rather than FOY or FUT, if there is no hemorrhagic tendency. When a patient with DIC develops cerebral infarction, one should always investigate for possible malignant tumors. PMID- 8703563 TI - [Magnetic resonance imaging of cavernous angioma of the brain stem]. PMID- 8703564 TI - [A 62-year-old man with familial parkinsonism with the onset at 24 years of the age]. AB - We report a right-handed 62-year-old man with early onset familial parkinsonism. The patient was well until 24 years of the age when he noted an onset of resting tremor in his right hand. During the next four years, he noted rigidity, bradykinesia, and difficulty in walking. He was seen in another hospital at 28 years of the age, where he received left pallidotomy. Rigidity on the left side showed marked improvement. He received right pallidotomy at age 30 years. He developed right hemiplegia after this surgery. He was admitted to our hospital in March, 1983 when he was 51 years of the age. He was treated with levodopa but improvement was rather of minor degree. He was transferred to another hospital, but his motor disturbance progressed slowly, and was admitted again to our hospital in November 1990. He had 6 siblings 4 of whom including himself suffered from parkinsonism. No consanguinity was noted in parents. On admission, he appeared chronically ill but the general physical examination was unremarkable. Neurologic examination revealed an alert and mentally sound man. Hasegawa dementia scale was 28.5/32.5. Upward gaze was slightly restricted (3/5). Cranial nerve examination revealed oculogyric crisis, apraxia of eyelid opening, masked face, and small voice. He was able to stand with support; his posture showed left ward leaning. He had right hemiparesis with moderate weakness. He showed marked bradykinesia and moderate rigidity in his left upper extremity. Fine postural tremor was noted in the left hand. Deep tendon reflexes were diminished in the upper extremities. No Babinski sign was noted. Pain sensation was somewhat diminished on the right side. Results of routine laboratory examination were unremarkable. Cranial CT scan revealed atrophy in the frontal lobe, particularly in the prefrontal area. In addition, MRI revealed T1-and-T2-low signal intensity lesions in the right ventral pallidal region and in the left ventrolateral thalamic-hypothalamic areas. He was treated with 600 mg of levodopa with benserazide and 22.5 mg of bromocriptine with mild to moderate improvement in his bradykinesia and rigidity. He was discharged in January 1991. His clinical course was complicated by intestinal obstruction in October, 1994. He was admitted to another hospital where he was operated on the obstruction on November 5, 1994. The sigmoid colon was markedly dilated but no mass was found. Postoperative course was uneventful until November 18, 1994 when he was found dead in his hospital room shortly after 4 am. The patient was discussed in neurological CPC, and the chief discussant arrived at the conclusion that the patient had young onset familial Lewy body-negative parkinsonism. Opinions were divided between Lewy body-positive familial Parkinson's disease and Lewy body negative young onset parkinsonism. Postmortem examination revealed aspiration pneumonia, which appeared to be the cause of his death, in the right lung. Neuropathologic examination revealed loss of malanized neurons in the substantia nigra and the locus coeruleus. In the substantia nigra, neuronal loss was particularly severe in the ventrolateral area. No Lewy bodies were seen. The dorsal motor nucleus of the vagal nerve was well preserved. Stereotaxic lesions involved bilateral thalamic areas. This patient appears to represent a case of autosomal recessive juvenile parkinsonism (AR-JP). Early onset, superb response to levodopa, sleep effect, and easy development of dyskinesias and motor fluctuations characterize AR-JP. The reason why this patient did not show these clinical features is probably bilateral sterotaxic surgeries. Particularly, the second surgery was complicated by right hemiparesis. His siblings who developed parkinsonism showed typical clinical features of AR-JP. PMID- 8703565 TI - Clifton R. Gaus, administrator of AHCPR. Interview by Jan Towers and Carole Jennings. PMID- 8703566 TI - Using an encounter form to develop a clinical database for documenting nurse practitioner primary care. AB - A clinical database will enable nurse practitioners to document and improve characteristics of their practices. This information is vital if nurse practitioners expect to achieve success in the changing U.S. health care system. This article describes the process of creating an encounter form and a related clinical database. Changes made in a family practice clinic that were derived from using a clinical database are presented. Additional uses for a clinical database, such as quality assurance, clinical evaluation, and clinical research, are discussed. PMID- 8703567 TI - Polypharmacy and the older adult. PMID- 8703568 TI - Stevens-Johnson syndrome. PMID- 8703569 TI - Rational antibiotic selection. PMID- 8703570 TI - The acute care nurse practitioner in collaborative practice. AB - Nurse-physician relationships remain, for the most part, hierarchical in nature. A hierarchical structure allows the person at the top, most notably the physician, the highest level of authority and power for decision making. Other health care providers are delegated various tasks related to the medical plan of care. One role of nonmedical health care providers, including nurses, is to support the medical plan of care and increase the productivity of physicians. Medical centers have house staff, usually interns and residents, who work collaboratively with the attending physicians in care delivery. At one medical center, a shortage of medical house staff for internal medicine prompted the development and evaluation of an alternative service. The alternative service utilized master prepared, certified nurse practitioners on a nonteaching service to provide care for selected types of medical patients. Physicians consulted with nurse practitioners, but retained decision-making authority concerning patient admission to the service. This paper describes the development and evaluation of an alternative service based on a collaborative practice model and the role of nurse practitioners working under such a model. Discussion includes suggestions for process guideline development for organizations that want to improve collaborative practice relationships between unit nursing staff, nurse practitioners, and physicians. PMID- 8703571 TI - The perceptions of Puerto Rican women regarding health care experiences. PMID- 8703572 TI - Fibromyalgia. PMID- 8703573 TI - Managing adult asthma in the primary care setting. PMID- 8703574 TI - Revisiting "a nurse for all settings": the nurse practitioner movement, 1965 1995. PMID- 8703575 TI - Morphometric analysis of noninvasively assessed bone biopsies: comparison of high resolution computed tomography and histologic sections. AB - This article describes a new method to analyze the structural behavior of trabecular bone structure by means of noninvasive measurements of the three dimensional cancellous bone architecture. For the noninvasive data acquisition, a high-resolution quantitative computed tomography system for peripheral measuring sites (pQCT) was used. With this system and the help of a multiple thin-slice measuring technique, it became possible to examine three-dimensional bone structure with a resolution of 0.25 mm. Using a special three-dimensional segmentation algorithm, mineralized bone was separated from bone marrow and muscle tissue within the three-dimensional stack of CT slices. These segmented data sets can then be processed nondestructively and, even more important, repetitively in either two or three dimensions. In order to validate this noninvasive procedure, a two-dimensional comparative morphometric study was performed including CT slices and corresponding histologic sections prepared after CT measurement. Three representative sections from the three-dimensional stack of CT slices were selected and the morphometric indices of the segmented CT slices were compared with the indices stemming from the corresponding histologic sections prepared after CT measurement. Although the presented approach can only give an example of the method, the results from the morphometric analysis support the assumption that cancellous bone structures based on noninvasive high resolution CT measurements are representative for trabecular microstructures assessed from histologic bone sections. The study demonstrates the potential of high-resolution CT imaging for in vivo applications of quantitative bone morphometry. This is especially true for repetitive follow-up measurements, which cannot be performed using histologic sections. Additionally, the method offers an easy access to the three-dimensional structure of trabecular bone, which is mandatory for the analyses of the anisotropic mechanical behavior of cancellous bone. PMID- 8703576 TI - Osteoclasts are not the major source of interleukin-6 in mouse parietal bones. AB - Interleukin-6 (IL-6) is produced by bone cells and has been shown to stimulate the proliferation of osteoclast progenitors. Which cells in bone produce IL-6 is controversial. This article tests the hypothesis that tartrate-resistant acid phosphatase-positive osteoclasts (TRAP + OC) in neonatal mouse parietal bones are the major source of IL-6. Bones were preincubated with indomethacin to decrease the number of TRAP + OC and the amount of IL-6 produced. Incubation with parathyroid hormone or prostaglandin E2 increased the number of TRAP + OC and the amount of IL-6 produced. Calcitonin and 17 beta-estradiol inhibited this increase in TRAP + OC but had no effect on IL-6 production. 1,25-dihydroxy-vitamin D3 also stimulated an increase in TRAP + OC number but did not cause increased IL-6 production. Both the endocranial and ectocranial membranes of these bones produced large amounts of IL-6. TRAP activity in extracts of endocranial membranes was 14-fold that of the ectocranial membrane and, histochemically, some TRAP + cells could be detected here. However, the ectocranial membranes produced more IL-6 than the endocranial membranes. We conclude that TRAP + OC are not a major source of IL-6 in this system. PMID- 8703577 TI - Decreased bone mineral density associated with early menopause progresses for at least ten years: cross-sectional comparisons between early and normal menopausal women. AB - To establish whether early onset of menopause carries an increased risk of osteoporosis, we compared the bone mineral density (BMD) of the second to fourth lumbar vertebrae (L2-4) between 18 women who had menopause before 43 years of age (early menopause group) and 19 women who had menopause after reaching 43 years of age (normal menopause group). Serum levels of calcium, phosphorus, calcitonin, intact parathyroid hormone, luteinizing hormone (LH), follicle-stimulating hormone (FSH), estradiol (E2), and alkaline phosphatase activity were measured, and urine samples were analyzed to derive calcium/creatinine, hydroxyproline/creatinine, pyridinoline/creatinine, and deoxypyridinoline/creatinine (D-Pyr/Cr) ratios. Mean BMD was significantly lower in the early menopause group than in the normal menopause group, and individual BMD values in about half of the subjects in the former group were below the fracture threshold for Japanese women. Serum concentrations of LH, FSH, and E2 were slightly, but not significantly, lower in the early menopause group than in the normal menopause group. The D-Pyr/Cr ratio was significantly higher in the early menopause group than in the normal menopause group. There was no correlation between L2-4 BMD and age or the number of years after menopause in the normal menopause group, but both age and the number of years after menopause were negatively correlated with L2-4 BMD in the early menopause group. These results indicate that BMD in women who have early menopause continues to decline for up to 10 years, and that menopause and aging increase the risk of osteoporosis. PMID- 8703578 TI - Effects of X-ray irradiation on terminal differentiation and cartilage matrix calcification of rabbit growth plate chondrocytes in culture. AB - The retardation of long bone growth caused by irradiation is thought to be closely related to the impairment of growth plate function, but its mechanism remains unclear. In this study, we examined the effects of irradiation on the terminal differentiation of growth plate chondrocytes and on calcification. Chondrocytes were isolated from the growth plate of the ribs of four-week-old rabbits and inoculated at a high density on a type-I collagen-coated dish. Following logarithmic proliferation, they reached confluence (premature chondrocytes), then matured (mature chondrocytes), and became hypertrophied (hypertrophic chondrocytes). 10 Gy or less irradiation of the premature chondrocytes potently inhibited the terminal differentiation and matrix mineralization. Irradiation inhibited chondrocyte hypertrophy and suppressed alkaline phosphatase induction and the expression of type-X collagen without changing the protein composition profile of any other cell layer. Premature cells had the highest radiosensitivity. The sensitivity was decreased as the cells differentiated; the effects of irradiation on hypertrophic chondrocytes with terminal differentiation-related phenotypes were reduced. This study showed that 10 Gy or less irradiation of growth plate chondrocytes impaired terminal differentiation and mineralization. Since chondroclasts and bone marrow cells invade only to the mineralized cartilage, the induction of calcification in cartilage matrices is one of the most important steps in endochondral ossification. Therefore, it is conceivable that the damage in the growth plate induced by irradiation could account for the subsequent abnormal bone and skeletal growth. PMID- 8703579 TI - Comparative study of bone mineral density estimated by various methods of single- and dual-energy quantitative computed tomography: the capability of the four equation four-unknown method. AB - A dual-energy (DE) quantitative computed tomography (QCT) method, the four equation four-unknown method (DEQCT 4E-4U), was assessed and compared to single energy (SE) QCT and standard DEQCT (two-line method). The results of this study indicate that bone mineral density (BMD) was more accurately estimated by the present method than by the SEQCT or standard DEQCT techniques on the basis of a phantom study when a large fat content was present. The results of both the phantom study and a human study also showed that the present method corrected for fat in estimating BMD in the presence of high-fat content. These findings suggest that use of this method for estimating BMD can provide useful information in studies assessing the metabolic state of bone. We propose that CT numbers estimated from excised vertebral bone marrow can serve as a soft-tissue correction for the present method. PMID- 8703580 TI - Vitamin D receptor polymorphism, bone mineral density, and osteoporotic vertebral fracture: studies in a UK population. AB - Bone mineral density is under strong genetic control and polymorphisms of the vitamin D receptor (VDR) have been suggested to account for some of the genetic variation in bone mass. However, the relationship between VDR polymorphisms and bone density is controversial and has not been confirmed by all workers. Since there is little information on the association between VDR genotype and bone mass in the UK, we studied VDR genotype, bone mineral density, and osteoporotic fracture in a cohort or pre- and postmenopausal women from the Northeast of Scotland. We found a highly significant "inverse" association between the VDR genotype and bone mineral density at the hip such that individuals of "bb" genotype had a femoral neck bone density of 0.79 standard deviation lower than individuals of BB genotype (p < 0.02). This contrasts with most previous studies in which the "bb" genotype has been associated with high bone density. A similar, but nonsignificant trend was seen for lumbar spine BMD. To study the clinical significance of this observation, we examined the distribution of VDR genotypes in a subgroup of patients with severe osteoporosis who had vertebral compression fractures (n = 44) as compared with with age-and gender-matched controls (n = 44). Despite the differences in BMD between genotypes, there was no significant excess of any specific VDR genotype in osteoporotic fracture patients, indicating that VDR genotyping may be of limited practical value in identifying patients at risk of vertebral fracture. This study confirms that there is a significant association between VDR genotype and bone mass in our population. The "inverse" relationship between VDR genotype noted in this, as compared with previous studies, would be consistent with a model whereby VDR polymorphisms are not the cause of reduced BMD, but rather, are in linkage disequilibrium with a disease causing locus nearby. PMID- 8703581 TI - Strontium increases vertebral bone volume in rats at a low dose that does not induce detectable mineralization defect. AB - Low doses of strontium and fluoride were shown to increase bone formation and trabecular bone density in rodents. To assess whether strontium or fluoride affect the quality of the mineral at doses known to increase bone density, we have determined the effects of low doses of strontium and fluoride on bone formation and bone mineral characteristics in rats. Adult rats were given strontium alone (0.20%), fluoride alone (1 mg/kg per day), or the combined treatment for 8 weeks. Strontium levels in serum and femur were similar in groups treated with strontium alone or in combination, being about 5% of calcium levels. Biochemical and neutron activation analyses in femur showed that calcium and magnesium contents did not differ in the four group of rats, suggesting that strontium was incorporated in the apatite lattice of the bone minerals in the strontium-treated rats. The mineralized bone volume was significantly increased by 17% in the strontium-treated group, by 20% in the fluoride-treated group, and by 19% in rats given with the combined treatment. This was associated with increased osteoid surface, osteoblast surface, and double tetracycline labeled surfaces in the strontium-treated and fluoride-treated groups, showing that the number of bone forming sites was increased. However, the mineral apposition rate, the osteoid thickness, and the mineralization lag time were similar in controls and treated groups, reflecting the lack of deleterious effects of low doses of strontium and fluoride on bone mineralization. The density fractionation analysis measured in the femur also showed that neither strontium, nor fluoride at the low doses used, significantly altered the mineralization profile. The results indicate that treatment with low doses of strontium or fluoride increase the number of bone forming sites and vertebral bone volume in rats, but does not have detectable adverse effects on the mineral profile, bone mineral chemistry or bone matrix mineralization. PMID- 8703582 TI - Ovulatory premenopausal women lose cancellous spinal bone: a five year prospective study. AB - Healthy premenopausal women with regular cycles are believed to be increasing or maintaining bone density. However, few studies have prospectively documented spinal cancellous bone, the bone that changes rapidly in response to reproductive hormones, in this population. Furthermore, our previous one-year study documented that 24% of the one-year bone change by quantitative computed tomography (QCT) was related to subclinical ovulatory disturbances (short luteal phase and non ovulation) in the presence of regular menstrual cycles. The purpose of this study was to document the cancellous bone change over five years in this initially ovulatory, premenopausal cohort of 66 healthy women. Thirty-seven women, who continued to be premenopausal and have regular cycles, completed this five-year study. Those enrolled differed only by being older and weighing less than those who could not be contacted (n = 19) or who declined to participate (n = 10). Documentation of current ovulatory characteristics was obtained for at least three cycles in 27 women. At the five-year assessment, the volunteers were 40.6 (range 26-47) years old, weighed 58.5 (41-77) kg, and were 160.9 (149-174) cm in height. All were premenopausal, healthy, nonsmokers with regular menstrual cycles (mean 27.7, range 24-33 days). Six women with intervening events (such as pregnancy or use of oral contraceptives) had interval (12 to 60 months) QCT changes similar to the remaining 31 (-7.98 vs. -4.92 mg/cm, p = 0.1, respectively). Mean five-year QCT was 143.0 +/- 20.2 mg/cm, whereas the initial mean value was 151.9 +/- 20.1 mg/cm. Significant QCT loss over five years (-8.9 +/- 6.2 mg/cm) (95% Cl -6.9 to -11.0) correlated with QCT change in the first year (r = 0.629, p < 0.001). First-year change was not related to the subsequent four-year interval change (r = -0.056, p = 0.74), however. Five-year QCT change was not related to age, weight, osteoporosis family history, estimated calcium intake, or exercise, but did correlate with year one luteal index (luteal/cycle length) (r = 0.339, p = 0.043). Significant cancellous spinal bone loss occurs in healthy, ovulatory premenopausal women, and is influenced by subclinical disturbances of ovulation. PMID- 8703583 TI - Characterization and function of the receptor for IGF-I in human preosteoclastic cells. AB - Using a coculture system, we have recently demonstrated that insulin-like growth factor I (IGF-I) is a mediator of preosteoclastic cell migration toward bone derived endothelial cells. To better characterize the mechanisms of IGF-I action on preosteoclastic cells we evaluated the expression of type I IGFs receptor in the human leukemic cell line, FLG 29.1, which differentiates toward the osteoclastic phenotype following phorbol ester (TPA) treatment. Scatchard analysis of 125I-labeled IGF-I to FLG 29.1 cells revealed the presence of a single high affinity binding site in both untreated and TPA-treated cells with a similar Kd value (0.3 +/- 0.2 nmol/L and 0.4 +/- 0.1 nmol/L, respectively). In untreated cells, IGF-I binding capacity (1.43 +/- 0.41 fmol/10(6) cells) was significantly (p < 0.05) lower than in TPA-treated cells (2.62 +/- 0.87 fmol/10(6) cells). Competition analyses and crosslinking studies revealed the presence of type I IGF receptor both in untreated and TPA-treated cells. Northern analysis demonstrated that mRNA for IGF-I receptor was expressed by both untreated and TPA-treated FLG 29.1 cells. In addition, FLG 29.1 cells released in the conditioned medium IGFBP-2 and IGFBP-4, whose expression was increased by TPA treatment as demonstrated by ligand and immunoblot analyses. The previous observations of chemotactic action of IGF-I on FLG 29.1 cells was confirmed by ultrastructural observations. Indeed, these cells revealed a marked migratory activity in response to nanomolar concentrations of IGF-I. In addition, the IGF-I receptor alpha IR-3 antiserum inhibited the IGF-I-induced FLG 29.1 cell's migratory activity. These findings clearly show that type IIGF receptor is expressed by osteoclast precursors and that IGF-I induces migration of these through the binding to type I IGF receptors. Binding proteins expressed by osteoclast precursors may play an autocrine role in modulating the IGF-I bioeffects. PMID- 8703584 TI - Modulation of the resorptive activity of rat osteoclasts by small changes in extracellular pH near the physiological range. AB - We investigated the effect of small shifts in extracellular pH on the resorptive activity of rat osteoclasts in vitro. Osteoclast-containing mixed bone cell populations disaggregated from neonatal rat long bones were cultured for 26 h at low density on bovine bone disks in HCO-3/CO2-buffered medium modified by the addition of small amounts of protons as HCl (2.5-15.0 mEq/L) or hydroxyl ions as NaOH (2.5-10.0 mEq/L). Little or no bone resorption occurred when ambient pH was above about 7.30; at pH 7.00, osteoclasts exhibited near-maximal activity. The greatest changes were associated with a pH difference of as little as 0.10 unit, between pH 7.25 and pH 7.15. Over this narrow range, the average number of resorption pits formed on each bone wafer increased sixfold. The steep sigmoidal pH response curve indicates that very slight alterations in ambient hydrogen ion concentration can effectively "switch on" or "switch off" rat osteoclasts in vitro. PMID- 8703586 TI - Bone metaplasia of urothelial mucosa: an unusual biological phenomenon causing kidney stones. AB - A kidney stone was analyzed by infrared spectrometry, by petrography using ground slices, and by undecalcified histology. Infrared spectrometry showed that the stone was composed of calcium oxalate monohydrate (whewellite), calcium oxalate dihydrate (weddellite), and calcium phosphocarbonate (apatite). Optical microscopy of the ground sections revealed tissue fragments within the stone containing small spaces resembling empty osteocyte lacunae. Histological analysis of methacrylate-embedded sections confirmed the presence of an area of heterotopic ossification. Urothelial mucosa is potentially capable of inducing bone formation and in this case the bony tissue had apparently functioned as a nucleation site for the calculus. PMID- 8703585 TI - Scurvy results in decreased collagen synthesis and bone density in the guinea pig animal model. AB - The effect of severe ascorbic acid deficiency on bone remodeling and collagen synthesis was evaluated in a 21 day experiment, using the scorbutic guinea pig model. Animals (n = 6-7/group) were assigned to one of three groups: scorbutic, pair-fed ascorbic acid-replete, or ad libitum ascorbic acid-replete groups. After 2 weeks, scorbutic animals started voluntarily decreasing food intake and losing weight. By day 19-21, at which time bone and tissue samples were collected and analyzed, scorbutic animals decreased food intake to 46% of usual and lost 9% body weight. Serum 25OHD3, 1,25(OH)2D3, calcium, and albumin were significantly lower (p < 0.05) in the scorbutic animals than in the other groups. Bone mineral density and bone mineral content of the proximal and central femur were significantly lower in the scorbutic group than in the other groups (p < 0.05). Morphometric analysis of tibia indicated significantly lower bone volume, fewer and thinner trabeculae, and a thinner growth plate in the scorbutic group, compared to the pair-fed and ad libitum groups (p < 0.05). Osteoclast surface was about 60% higher in the scorbutic group than in the pair-fed and ad libitum control groups (0.05 < p < 0.10). Mechanical strength of the femur and lumbar vertebral body tended to be lower when bone mass was altered in the same group. Collagen synthesis of articular cartilage and tendons was lower in the scorbutic group than in the pair-fed or ad libitum groups (p < 0.05). In conclusion, scurvy but not food restriction, per se, results in alterations in bone mass and tissue collagen synthesis. PMID- 8703587 TI - Who's representing whom? PMID- 8703588 TI - Good oral health? PMID- 8703589 TI - Hepatitis B status of applicants. PMID- 8703590 TI - Migraine attacks. PMID- 8703591 TI - An unusual radiograph finding. PMID- 8703592 TI - General anaesthesia. PMID- 8703593 TI - The role of the community dental practitioner. PMID- 8703594 TI - Specialist orthodontics in the general dental services. PMID- 8703595 TI - Crisis in orthodontic departments. PMID- 8703596 TI - Evidenced based orthodontics. PMID- 8703597 TI - X-ray microtomography: a novel technique for the quantification of effects in enamel following CO2 laser application. AB - X-ray microtomography (XMT) is a miniaturised version of the technique of computer axial tomography as used in medical diagnosis. As an example of the dental research applications of the technique, a pilot study is described in which the effects of a continuous wave carbon-dioxide laser on dental enamel are investigated and the use of XMT for the non-invasive measurement of mineral density changes within dental hard tissues is demonstrated. The results show clearly the ability of XMT to display graphically, and to quantify, changes in mineral density occurring within lased specimens, compared with a conventional SEM view of similar material. There have been many reports of the problems found when using lasers to cut dental hard tissues, and this paper demonstrates that XMT is of particular value in the study of the cracking induced by many lasers. We also suggest that XMT might be used in many studies investigating mineral density changes within dental hard tissues. PMID- 8703599 TI - Mucosal lesions due to oral cocaine use. AB - Cocaine abuse is a major public health problem worldwide and is increasingly common in Europe. Oral use of cocaine in two patients resulted in unusual white lesions on their anterior maxillary gingivae/vestibule. GDPs should be alert to this newly described lesion. PMID- 8703598 TI - Lingual nerve damage during lower third molar removal: a comparison of two surgical methods. AB - The high incidence of lingual sensory disturbance following lower third molar removal in the UK may be due to the elevation of a lingual flap and insertion of a Howarth's periosteal elevator, in an attempt to protect the lingual nerve. We have therefore studied the validity of this technique by recording the incidence of temporary and permanent lingual nerve injury during 771 operations randomly allocated to be carried out with or without lingual flap retraction. Surgery with lingual flap retraction resulted in lingual sensory disturbance in 6.9% and this persisted, requiring lingual nerve repair, in 0.8%. Surgery without lingual flap retraction resulted in lingual sensory disturbance in 0.8% (P < 0.0001) and this persisted, requiring lingual nerve repair, in 0.3%. We conclude that avoidance of lingual retraction reduces the incidence of temporary lingual nerve disturbance and does not increase the incidence of permanent damage. This indicates that use of the Howarth's in this way is invalid, and suggests that for the majority of cases, lingual retraction should be avoided. PMID- 8703600 TI - An unusual case of trismus and dysphagia. AB - At one time the incidence of broken hypodermic needles was a fairly common occurrence in general dental practice. Fortunately the routine use of modern stainless steel needles has all but eliminated this emergency. Occasionally a patient may make a sudden movement during administration of local anaesthetic, particularly in the provision of an inferior dental block. A case is described where this occurred with subsequent fracture of the hypodermic needle. The patient experienced sudden trismus and dysphagia as the needle fragment became deeply impacted in the surrounding soft tissues. PMID- 8703601 TI - Practical marketing for dentistry. 3. Relationship marketing and patient/customer satisfaction. AB - In this article, we look at the philosophy of customer focus and value, and how dental practices can produce and deliver high customer value and satisfaction, to retain as well as attract their customers-the patients. Total quality concepts will also be discussed in the context of their relationship with marketing activities. In all cases, where 'customer' is referenced, this means 'patient' in the context of a dentistry, since patients are the customers, their requirements must be considered in targeting the marketing of a dental practice. PMID- 8703602 TI - Hepatitis C virus (HCV)--an occupational risk to dentists? AB - Hepatitis B virus has long been regarded as an occupational hazard in dentistry. However, as the availability of effective vaccination has reduced the risk of HBV infection, so dental staff have become aware of the potential risk of transmission of hepatitis C virus during dental treatment. This article outlines current relevant data suggesting that nosocomial transmission of HCV in dentistry is unlikely. PMID- 8703603 TI - The nurse's role in diagnosing and treating onychomycosis. AB - Onychomycosis involves fungal invasion of the nail unit via the nail bed or nail plate. Nurses play an important role in diagnosing the disease, managing it, and educating patients about it. Newer oral antifungal agents have increased success rates with shortened treatment times. PMID- 8703604 TI - Photochemotherapy (PUVA) protocol. AB - PUVA is used to treat severe skin problems that have not responded well to more conventional forms of treatment. The effectiveness of PUVA therapy depends on patient compliance and careful administration and evaluation of patient response to treatment. The following protocol was developed to identify important factors to consider when administering PUVA therapy. PMID- 8703605 TI - What's your assessment? Allergic contact dermatitis to hair dye. PMID- 8703606 TI - Is a tan really beautiful? AB - History has demonstrated many dangerous beauty fads, including tanning, which have evolved into a false concept of beauty. A teaching tool, which emphasizes tanning as just a fashion trend rather than a beauty enhancer, can help discourage people from suntanning their skin. PMID- 8703607 TI - The skin, free radicals, and oxidative stress. AB - Oxidative stress involves the adverse effects of oxygen and other free radicals on living tissue. An overview of the various types of free radicals generated in the body, the basic chemistry of free radicals, and how they arise is presented. The importance of understanding the action of free radicals on specific target tissues in the skin and how this affects the physiology of the skin in relation to treatment is covered. PMID- 8703608 TI - Localized bullous pemphigoid occurring in a surgical wound. AB - Localized forms of bullous pemphigoid have been reported to occur in sites of trauma. The case of a 79-year-old female who developed blisters at the site of an abdominal surgical wound is reported. The diagnosis of bullous pemphigoid was made by histology and immunofluorescence studies. The reported cases of localized bullous pemphigoid associated with surgical wounds are reviewed. PMID- 8703609 TI - What's your assessment? Dyshidrotic eczema. PMID- 8703610 TI - Social support and health promotion outcomes of adults with psoriasis. AB - Illness and health behavior, social support, and health promotion outcomes (time of recovery from an acute illness episode and psychosocial adjustment to illness) were investigated in 44 adults with psoriasis. Illness behavior was significantly associated with shortened recovery time. PMID- 8703611 TI - Gloves prove beneficial. PMID- 8703612 TI - Epidermolysis bullosa: practical management and clinical update. AB - Epidermolysis bullosa (EB) is a heterogeneous group of rare, heritable disorders characterized by marked fragility of the skin and mucosa. Patients require a specialized program of wound care and timely intervention to treat the many systemic complications, some of which are potentially life-threatening. PMID- 8703613 TI - The influence of dressings on the costs of wound treatment. AB - To calculate cost effectiveness of wound care, it is incorrect to calculate the costs of just a dressing. All aspects influencing the costs of treatment should be calculated. These include costs for the actual materials, labor, and nonmedical costs to society. PMID- 8703614 TI - Nephrology teaching record. PMID- 8703615 TI - The Manitoba Nephrology Nursing Course. PMID- 8703617 TI - Reflections on the 1995 CANNT national symposium: an open letter to the nephrology nurses in Canada. PMID- 8703616 TI - The pre-dialysis clinic at the Toronto Hospital--is it working? PMID- 8703618 TI - Multiple organ failure: from basic science to prevention. PMID- 8703619 TI - The immuno-inflammatory cascade. PMID- 8703620 TI - Sepsis and cytokines: current status. AB - Sepsis is a constellation of clinical signs and symptoms resulting from excessive systemic host inflammatory response to infection. This inflammatory response is largely mediated by cytokines, which are released into the systemic circulation. Plasma concentrations of specific cytokines, TNF alpha, IL-1 beta, IL-6 and IL-8 are frequently elevated in human sepsis and cytokine concentrations correlate with severity and outcome of sepsis. In addition to pro-inflammatory cytokines, soluble cytokine receptors, cytokine receptor antagonists and counter inflammatory cytokines are also produced in large quantities in patients with sepsis; however, the specific role of these molecules in sepsis remains undefined. A complex interaction of cytokines and cytokine-neutralizing molecules probably determines the clinical presentation and course of sepsis. Intervening in this sequence of events to modify the host inflammatory responses may prove to be a beneficial treatment strategy for sepsis, but currently tested anticytokine therapies have been largely unsuccessful. PMID- 8703621 TI - Nutrition and immunonutrition. PMID- 8703622 TI - Pre-emptive analgesia with NSAID--what does it achieve? PMID- 8703623 TI - Cytokine balance and immunosuppressive changes at cardiac surgery. PMID- 8703624 TI - Perioperative changes in alpha 1-acid glycoprotein concentrations. PMID- 8703625 TI - CBF in adults using near infrared spectroscopy (NIRS): potential for bedside measurement? PMID- 8703626 TI - Hypercoagulability induced by crystalloids. PMID- 8703627 TI - Disturbance of leucocyte circulation and adhesion to the endothelium as factors in circulatory pathology. PMID- 8703628 TI - Mechanisms of cell injury and death. PMID- 8703629 TI - Molecular mechanisms of drug metabolism in the critically ill. PMID- 8703630 TI - Determinants of splanchnic blood flow. PMID- 8703631 TI - Splanchnic ischaemia/reperfusion and multiple organ failure. PMID- 8703632 TI - Determinants of intestinal barrier failure in critical illness. PMID- 8703633 TI - Mechanisms of acute renal failure. PMID- 8703634 TI - Fulminant and late onset hepatic failure. PMID- 8703635 TI - New approaches to acute lung injury. PMID- 8703636 TI - Patterns of HIV/AIDS in Zimbabwe: implications for health education. AB - Current patterns of HIV/AIDS in Zimbabwe are examined using national surveillance data reflecting four main variables: age, gender, marital status, and residence. The data show that the distribution of cases by age and gender is consistent with the literature on the rest of sub-Sahara Africa. Distribution of cases by age is bimodal with incidence highest among sexually active young adults (20-39 years) and the very young (0-4 years), and the male-to-female ratio is nearly 1:1 among adults. However, among adolescents 15-19, the incidence among females is six times greater than their male counterparts. The data also show that married persons have the highest rate of HIV infection (51%), greater than single persons (34%). These patterns highlight the significance of heterosexual and perinatal transmission. These data suggest a need for expanded HIV testing, counseling, and education across gender and marital status in order to detect HIV infection earlier in the spectrum of the disease. Promotion of monogamy and proper condom use should be a major focus of the empowerment process. Further recommendations are provided for planning, implementation, evaluation, and coordination of a comprehensive national prevention strategy. PMID- 8703637 TI - AIDS education for college students: review and proposal for a research-based curriculum. AB - The history of AIDS education for college students in the U.S. is reviewed. Wide agreement concerning the goals and overall content of the AIDS-education curriculum is found. However, the context and location of AIDS education within the curriculum vary considerably. Increased knowledge about AIDS is a frequent outcome, but improvements in attitude and reported sexual behaviors are more difficult to achieve. The conceptual frameworks used in AIDS education include the health belief model, social learning theory, and theory of reasoned action; each has contributed to the design of an AIDS-prevention curriculum. A proposed research-based AIDS-education curriculum would utilize the theory of planned behavior from attitude research, the elaboration likelihood model from persuasion research, and the conceptual change model from science education research. PMID- 8703639 TI - Mental health-related calls to the CDC National AIDS Hotline. AB - The CDC National AIDS Hotline provides confidential HIV-related information and referrals to anonymous callers, twenty-four hours a day. As part of a continuing quality improvement assessment of caller informational needs, 302 randomly selected anonymous overnight calls to the Hotline were evaluated for mental health-related content. Of 302 calls, 34 calls (11.3%) were mental health related, in that callers spoke about specific mental health-related topics or requested mental health referrals, and 14 calls (4.6%) included signs or symptoms from the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-IV) potentially indicative of mental illness. The results suggest that training in recognizing and referring mental health calls might be useful for Hotline workers. The results also serve as a reminder for all health care professionals and organizations of the potential for overlap between patients' mental health-related needs and patients' HIV-related needs. PMID- 8703638 TI - What African-American women know, do, and feel about AIDS: a function of age and education. AB - Because African-American women tend to be studied as a homogeneous group, little data exist that report differences in what they know, feel, and do about AIDS. The purpose of this study was to compare African-American women across two age groups and four educational groups to discern if differences exist. African American women who were 20 years of age or over were recruited in a variety of settings to obtain a heterogeneous sample. The sample totaled 281 women. Participation consisted of the completion of the AIDS Knowledge, Feelings, and Behavior Questionnaire. Multivariate analysis of variance revealed that educational level influenced their knowledge and attitudes about AIDS, whereas age influenced not knowledge but attitudes and sexual behavior. Based on race alone, African-American women should not be treated as homogeneous in what they know, do, and feel about AIDS. PMID- 8703640 TI - Childhood sexual abuse linked with adult substance use, victimization, and AIDS risk. AB - College women who report childhood sexual abuse were compared with women who do not report abuse on a number of variables concerned with problems in living. Multivariate Analysis of Variance revealed that, compared with nonabused women, sexually abused women reported significantly more negative attitudes about sexuality, less sexual assertiveness about birth control or refusing unwanted sex, less efficacy concerning HIV prevention, more anticipation of a negative response from a partner concerning safer sex, more hard-substance use, and more sexual victimization in adulthood. These results support and extend previous work in this area and argue for greater attention to relational issues for interventions with sexually abused women. Limitations to the study and future directions for research are discussed. PMID- 8703641 TI - Compliance with AZT treatment regimen of HIV-seropositive injection drug users: a neglected issue. AB - Although treatment with zidovudine (AZT) is now recommended for asymptomatic and symptomatic HIV-infected persons with CD4+ cell counts of 0.20 to 0.50 x 10(9)/L and under, data gathered from a small convenience sample of current and former injection drug users with AIDS in the New York City metropolitan region suggest that noncompliance with HIV/AIDS-related therapeutic regimen may be common in this population. This paper enumerates the reasons for noncompliance offered by these informants, reviews the general literature on treatment compliance to identify additional potential reasons for non-adherence to AZT treatment regimen, and outlines some suggestions for future research into this important issue that may prompt changes in the antiviral delivery system. PMID- 8703643 TI - My guardian angel: a learning exercise for adolescent AIDS education. AB - This article describes the methods to conduct a simple role-play exercise. The purpose of the exercise is to practice skills to prevent HIV infection and AIDS, including peer communication about personal health decisions among high school and college students. With guidance from the instructor, each student individually prepares a written Personal Prevention Plan for actions they intend to prevent AIDS. Following completion of the Plan, two pairs of students role play using a prepared scenario. The scenario includes a dilemma about a dating couple choosing to have sexual intercourse and the reactions of their closest friends. Other students in the class observe the interactions of their peers. After the role play, all of the students discuss factors that influence dating and intimate behaviors including peer attitudes and beliefs. Students volunteer ideas to counter perceived peer pressure for sexual intercourse. PMID- 8703642 TI - Correlates of acceptance of HIV testing and post-test counseling in the obstetrical setting. AB - Recent results of AZT testing among pregnant women by NIH indicate that early diagnosis and medical intervention can reduce vertical transmission of HIV. Debates have been rekindled concerning whether testing should remain voluntary or be made mandatory. This article analyzes predictors of women's decisions to accept testing voluntarily and return for their test results. Although derived from the postpartum setting, this information is also likely to be useful in understanding voluntary acceptance of prenatal HIV testing, as well as acceptance among those who are hard to reach during pregnancy. Results indicate that the time spent counseling each client and the individual HIV counselor were the best predictors of which women were most likely to test. Minority, self-paid, or uninsured clients, and women with little prenatal care were least likely to return for post-test counseling. The findings of this study point to the central importance of the counselor and the counseling process and to the relatively lesser impact of patient characteristics. Further study of counseling content and counselor performance is recommended. PMID- 8703644 TI - Cardiorespiratory effects of continuous i.v. administration of the ACE inhibitor enalaprilat in the critically ill. AB - 1. Cardiorespiratory effects of long-term, continuous i.v. administration of the ACE inhibitor enalaprilat were studied. 2. Forty-five consecutive critically patients suffering from trauma or postoperative complications were randomly separated into three groups (15 patients in each group) receiving either 0.25 mg h-1 or 0.50 mg h-1 enalaprilat, respectively, or saline solution as placebo (= control group). The infusion was continued for 5 days. 3. Haemodynamic and respiratory parameters were intensively monitored on admission to the intensive care unit (= 'baseline' values) and daily during the next 5 days. 4. Mean arterial blood pressure (MAP) decreased significantly only in the enalaprilat treated patients, whereas heart rate (HR) remained unchanged in these patients. 5. Pulmonary capillary wedge pressure (PCWP) and pulmonary artery pressure (PAP) were decreased by enalaprilat (0.50 mg h-1: PAP (mean +/- s.d.) decreased from 28.0 +/- 4.1 to 24.0 +/- 3.0 mm Hg) and remained significantly lower than in the control group. In the untreated control group, cardiac index (CI), oxygen consumption (VO2I) and oxygen delivery (DO2I) significantly decreased, which was blunted by enalaprilat infusion. Oxygen extraction (O2-extr) increased in both enalaprilat groups (0.25 mg h-1: from 26.1 +/- 5.5 to 30.4 +/- 4.0%; 0.50 mg h-1: 25.2 +/- 5.6 to 30.9 +/- 4.4%) and decreased in the control patients. 6. Right ventricular haemodynamics improved by enalaprilat infusion (0.50 mg h-1: RVEF increased from 40.0 +/- 3.5 to 45.5 +/- 4.0%). Lactate plasma concentrations decreased in the group with 0.50 mg h-1 enalaprilat (from 1.9 +/- 1.0 to 1.3 +/- 0.3 mg dl-1) and increased in the control patients. 7. Continuous infusion of the ACE inhibitor enalaprilat exerted beneficial cardiorespiratory effects in the critically ill. The widespread common risk of altered perfusion with decreased CI, DO2, VO2, O2-extr and increased lactate concentration was blunted by enalaprilat infusion. 8. Although 0.5 mg h-1 enalaprilat was most effective, a dose of 0.25 mg h-1 also showed beneficial haemodynamic effects in the critically ill. PMID- 8703645 TI - Resolution of ACE inhibitor cough: changes in subjective cough and responses to inhaled capsaicin, intradermal bradykinin and substance-P. AB - 1. In eight hypertensive patients with ACE inhibitor-induced cough the resolution of the cough was examined in a prospective observational study over 4 weeks duration. Resolution of cough was measured by visual analogue scales and questionnaire at baseline and days 3, 7, 14 and 28. In addition changes in cough sensitivity to inhaled capsaicin, and skin responses to bradykinin and substance P were measured at the same time points. 2. All patients recorded significant subjective improvement in cough questionnaire scores for severity and night time waking, and by visual analogue scales for severity and frequency of cough (all P < 0.0005 for trend from day 0-28). Significant changes in subjective measures were recorded by 3 to 7 days for most measures, but further reductions were observed up to day 28 (all P < 0.01 day 28 vs day 0). 3. The sensitivity to inhaled capsaicin fell over the 28 days of study after stopping enalapril. The potency of capsaicin relative to day 0 was reduced to 0.25 (95% CI 0.07-0.87) by day 14, and to 0.20 (95% CI 0.06-0.67) by 28 days. 4. After stopping enalapril there was a highly significant reduction in wheal area produced by intradermal bradykinin, with the majority of the effect seen by day 3 (P < 0.0005). The wheal area to intradermal substance-P also declined with time after stopping enalapril, but significant changes were not observed until 14 days (P < 0.01). 5. Multiple regression analysis of the rates of decline for the subjective and objective measures of cough showed significant associations between the response to inhaled capsaicin and the VAS scores for severity of cough (P = 0.005) and frequency of cough (P = 0.011). Capsaicin response was not related significantly to the severity of cough measured by self-administered questionnaire. 6. There was a significant association between bradykinin response and VAS scores for frequency of cough (P < 0.04) and severity of cough (P < 0.05), but not with cough by questionnaire or the capsaicin response. The response to substance-P did not relate significantly to any of the measures of cough. 7. Cough caused by enalapril improved markedly by 14 days but took up to 28 days to resolve. It was associated with increased sensitivity to inhaled capsaicin which decreased over 28 days, and which paralleled changes in subjective cough scores. PMID- 8703646 TI - Effects of the angiotensin type I receptor antagonist, losartan, on systemic and regional vascular responses to lower body negative pressure in healthy volunteers. AB - 1. The effects of a single oral dose (50 mg) of the angiotensin II AT1-receptor antagonist, losartan, on the systemic and regional vascular responses to simulated orthostatic stress by the lower body negative pressure (LBNP) technique were investigated in nine healthy volunteers, in a double-blind, placebo controlled crossover study. 2. Arterial blood pressure remained unchanged throughout the study. Three hours after its administration and before LBNP, losartan selectively increased renal blood flow (PAH clearance) by 8.3% (3.5 to 13.1%, 95% CI) from 1.25 +/- 0.08 l min-1 (P < 0.05) and decreased plasma aldosterone levels by 58% (29 to 87%, 95% CI) from 22 +/- 3 ng 100 ml-1 (P < 0.05). 3. LBNP at -10 and -20 mm Hg induced a progressive and significant decrease in central venous pressure and increases in forearm (plethysmography) and splanchnic (indocyanine green clearance) vascular resistances which were similar after losartan and placebo administrations. Losartan blunted the LBNP induced increase in renal vascular resistance observed at -20 mm Hg after placebo but a similar increase in glomerular filtration rate (inulin clearance) was observed at LBNP -10 and -20 mm Hg after losartan and placebo. Calculated filtration fraction increased after placebo (LBNP -10 mm Hg) and losartan (LBNP 20 mm Hg). Finally, LBNP-induced changes in biological parameters were similar after losartan and placebo at all levels of LBNP. 4. Thus, losartan does not interfere with the adaptive forearm and splanchnic vascular responses and preserves renal haemodynamics during orthostatic stress simulated by LBNP in healthy volunteers. PMID- 8703647 TI - Ceftazidime pharmacokinetics in preterm infants: effect of postnatal age and postnatal exposure to indomethacin. AB - 1. The effects of postnatal age and postnatal exposure to indomethacin on the pharmacokinetic parameters of ceftazidime (CAZ) were investigated in 23 preterm infants (gestational age 28.7 +/- 1.7 weeks; weight 1086 +/- 311 g) on day 3 and day 10 after birth. 2. CAZ (25 mg kg-1) was administered by intravenous bolus injection. Blood samples were drawn from an arterial catheter at 0, 0.5, 1, 2, 4, 8, and 12 h after the dose and CAZ concentrations in serum were determined by h.p.l.c. CAZ pharmacokinetics followed a one-compartment open model. 3. The glomerular filtration rate (GFR) of all infants was studied by means of the 24 h continuous inulin infusion technique. 4. The total body clearance of CAZ (34.7 +/ 9.2 vs 50.6 +/- 19.6 ml h-1, P < 0.05; 30.7 +/- 5.9 vs 41.6 +/- 9.0 ml h-1 kg-1, P < 0.05) and GFR (0.72 +/- 0.11 vs 0.91 +/- 0.15 ml min-1, P < 0.05) increased, whereas the apparent volume of distribution (425 +/- 147 vs 352 +/- 108 ml, P < 0.05; 363 +/- 59 vs 292 +/- 44 ml kg-1, P < 0.005) and the elimination half-life (8.7 +/- 2.8 vs 5.0 +/- 0.9 h, P < 0.005) decreased significantly between day 3 and day 10 after birth. Clearance of CAZ increased with increasing GFR (r = 0.81, P < 0.001). 5. In infants with postnatal exposure to indomethacin the changes in CAZ pharmacokinetics were markedly reduced. 6. These results indicate that the dosage regimen of CAZ should be adjusted after the first week of life except in infants who were postnatally exposed to indomethacin. PMID- 8703648 TI - Comparison of the effects of desipramine on noradrenaline- and methoxamine-evoked venoconstriction in man. AB - 1. The dorsal hand vein compliance technique was used to investigate the dual effect of tricyclic antidepressants at the noradrenergic synapse (i.e. noradrenaline uptake blockade leading to potentiation and alpha 1-adrenoceptor blockade leading to antagonism of the effect of noradrenaline). The effects of a single oral dose (100 mg) of desipramine (DMI) on venoconstrictor responses to locally infused noradrenaline and methoxamine, a selective alpha 1-adrenoceptor agonist with little affinity for the uptake mechanism, were examined. 2. Eight healthy male volunteers participated in four weekly experimental sessions. Each session was associated with one of the following treatment conditions: noradrenaline/DMI, noradrenaline/placebo, methoxamine/DMI, methoxamine/placebo. Subjects were allocated randomly to treatments and sessions according to a double blind balanced design. Noradrenaline acid tartrate (0.33-33 ng min-1) and methoxamine hydrochloride (0.0135-135 micrograms min-1) were infused into the superficial dorsal hand vein; each dose was infused for 5-7 min with 5 min intervening washout periods. Systolic and diastolic blood pressure and pulse rate were recorded before the infusion and immediately after the infusion of the highest dose. Salivation, an index of anticholinergic activity of the antidepressant, was measured by the dental roll technique. 3. Both noradrenaline and methoxamine produced dose-dependent venoconstriction: the geometric mean ED50 for noradrenaline was 4.41 ng min-1 and for methoxamine was 2558 ng min-1; the potency ratio (noradrenaline/methoxamine) was 2884. DMI shifted the dose-response curve for noradrenaline to the left (ANOVA: P < 0.025), resulting in a dose-ratio of 0.26. DMI did not affect the dose-response curve for methoxamine significantly; the dose ratio was 1.24. 4. None of the local infusions and/or systemic treatments had any significant effects on supine systolic and diastolic blood pressure and pulse rate. 5. DMI caused a substantial (47.6%) reduction in salivary output that significantly differed from the slight statistically insignificant increase (5.8%) of salivary output recorded after placebo. 6. These results show that a single oral dose (100 mg) of DMI causes significant potentiation of the response to noradrenaline without significantly affecting the response to methoxamine. The potentiation is likely to be due to uptake blockade since the response to methoxamine was not affected. Furthermore, the lack of significant antagonism of the response to methoxamine indicates that a single oral dose (100 mg) of DMI does not cause sufficient alpha 1-adrenoceptor blockade to be detected as a pharmacodynamic change in our test system. PMID- 8703649 TI - beta-Adrenergic receptors contribute to hypoxaemia induced vasodilation in man. AB - 1. It was the aim of the present study to investigate the role of the beta adrenergic receptor in hypoxaemia induced vasodilatation in the human forearm. 2. The study was performed in 12 non-smoking male volunteers. In six subjects the local vascular effects of intra-arterially (i.a.) infused propranolol (0.1 mu kg 1 min-1) was determined during normoxaemia and hypoxaemia (peripheral oxygen saturation; SpO2 80%), and compared with the contra-lateral (control) arm. beta adrenergic receptor blockade by propranolol was confirmed by i.a. infusions of adrenaline. In six other subjects the effects of incremental hypoxaemia (SpO2 90, 85, 80%) on forearm- and finger blood flow was investigated. A difference between these vascular beds is the absence of vascular beta-adrenergic receptors in the finger. Forearm- and finger blood flow were measured by venous occlusion plethysmography. Plasma levels of (nor-)adrenaline were determined in both arterial and venous blood samples. Hypoxaemia was attained by gradual decompression of a hypobaric chamber and individually adjusted. 3. During normoxaemia the single infusion of propranolol did not influence forearm vascular resistance. In contrast, during hypoxaemia a net vasoconstriction was observed in the arm treated with propranolol, which was significantly different (P = 0.009) from the vasodilatation in the control arm (mean difference in response 40%; 95% confidence interval 13.2-66.8). The net arterio-venous spillover of noradrenaline from the forearm increased after the first 15 min of hypoxaemia (P < 0.05) and returned to baseline in the next 15 min. Arterial and venous plasma levels of adrenaline during hypoxaemia remained unchanged compared with normoxaemia. In the second set of experiments incremental levels of hypoxaemia induced a vasoconstriction in the finger, which was significantly different (P = 0.025) from the vasodilatation in the forearm (mean difference in response 300%; 95% confidence interval 63-537). 4. The data indicate that beta-adrenergic receptors contribute to hypoxaemia induced vasodilatation, despite unaltered adrenaline plasma concentrations. PMID- 8703650 TI - Comparative wheal and flare study of mizolastine vs terfenadine, cetirizine, loratadine and placebo in healthy volunteers. AB - 1. Mizolastine, a new benzimidazole derivative with potent selective, non sedative H1-histamine antagonist activity was compared with terfenadine, cetirizine and loratadine using the histamine-induced wheal and flare model in healthy volunteers. 2. Study design was a five way double-blind crossover design using a single dose of mizolastine 10 mg, terfenadine 120 mg, cetirizine 10 mg, loratadine 10 mg and placebo. 3. Histamine tests were performed on 10 occasions up to +24 h after dosing using an intradermal injection of histamine 2 micrograms with concommittant contralateral injection of a saline control. 4. Mizolastine, terfenadine, cetirizine and loratadine significantly (P < 0.001 vs placebo) inhibited the wheal and flare formation starting 1 to 2 h after dosing up to 24 h after dosing. 5. Mizolastine was significantly more active than loratadine on the wheal (P < 0.01) and flare (P < 0.05) inhibition from 3 up to 6 and 8 h respectively, as active as terfenadine on both parameters and as active as cetirizine on wheal inhibition while less active (P < 0.01) than cetirizine on flare inhibition at 2 and 12 h post-dosing. PMID- 8703651 TI - Patterns of drug treatment of schizophrenic patients in Estonia, Spain and Sweden. AB - 1. Patterns of drug treatment and the use of polypharmacy in schizophrenic in patients were compared and evaluated in the University Teaching Hospitals of Psychiatry in Badajoz, Spain, Huddinge, Sweden, and Tartu, Estonia. 2. The medical records of up to 100 consecutively admitted patients were retrospectively reviewed using a standardized data form. 3. The male patients were significantly younger than females in all study locations, but there were no age differences between the locations. The length of stay was equal for the two series in the same hospital, but considerably longer in Tartu than in Badajoz and Huddinge. 4. The neuroleptic drugs used most commonly in Badajoz and Tartu were similar in prescription frequency and in the doses prescribed, but different from those used in Huddinge. Haloperidol was the most frequently prescribed neuroleptic in Badajoz and Tartu, accounting for one third of all neuroleptic prescriptions. In Huddinge the choice of neuroleptics was more evenly spread over several compounds. Intramuscular injections other than depot preparations were commonly used in Tartu and Badajoz, but not in Huddinge. 5. At least two neuroleptics were prescribed simultaneously on 73% of treatment days in Badajoz and 46% in both Huddinge and Tartu. The average cumulative daily doses of concomitant multiple neuroleptic treatment, expressed in chlorpromazine equivalents, were lower in Huddinge than in the other study locations and higher for male patients in Badajoz and Tartu. 6. Anticholinergics were used together with neuroleptics in 42% of treatment days in Badajoz and 30% in Huddinge as compared with 75% in Tartu. The use of anticholinergics increased in parallel to the increase in the number and the cumulative dose of concomitant neuroleptics in all study locations. 7. About 15% of patients in Badajoz and Tartu, but only 1% in Huddinge, received concomitant treatment with antidepressant drugs. The simultaneous use of antidepressants and benzodiazepines was inversely related to the number and the cumulative dose of neuroleptics in Badajoz and Tartu. In contrast, the cumulative dose and number of neuroleptics were greater, when additional benzodiazepines were prescribed in Huddinge. 8. The study in schizophrenic in-patients revealed that polypharmacy with concomitant multiple neuroleptics, additional anticholinergics and other psychotropics is an international phenomenon. PMID- 8703652 TI - Plasma hydroxy-metronidazole/metronidazole ratio in patients with liver disease and in healthy volunteers. AB - Metronidazole pharmacokinetics were studied in patients with different degrees of liver cirrhosis, classified according to the Child-Pugh algorithm (A, B or C, as liver disease severity increases) and in schistosomic patients. Metronidazole (500 mg) was administered i.v. as a slow infusion over 20 min, and blood samples were collected at set intervals after the end of the infusion. The plasma concentrations of metronidazole and its main metabolite hydroxy-metronidazole were quantified by reversed-phase h.p.l.c. with u.v. detection. The metronidazole and hydroxy-metronidazole areas under the curve from 0 to 24 h (AUC0,24h), the metronidazole terminal elimination half-life (t1/2), the total clearance (CL), the metronidazole volume of distribution (V) values and the hydroxy metronidazole/metronidazole concentration ratios as a function of time were calculated for each group. Comparison of the metronidazole AUC0,24h, t1/2 and CL values revealed that metronidazole metabolism is progressively impaired as the severity of liver disease increases. There were no variations in these parameters between the schistosomic and Child-Pugh A groups. In addition, there were no differences in the V and hydroxy-metronidazole AUC0,24h among the various groups studied. However, metronidazole metabolism was delayed in patients with hepatic disease, as illustrated by the hydroxy-metronidazole/metronidazole ratio 10 min after the end of metronidazole infusion. These results indicate that the clinical assessment of liver disease is paralleled by an impairment of metronidazole metabolism. Of the studied variables, we propose the hydroxy metronidazole/metronidazole ratio 10 min after metronidazole infusion as a suitable and practical index for liver function evaluation. PMID- 8703653 TI - The effects of selective serotonin reuptake inhibitors and their metabolites on S mephenytoin 4'-hydroxylase activity in human liver microsomes. AB - The inhibitory effects of four selective serotonin reuptake inhibitors (SSRIs), fluoxetine, sertraline, paroxetine and citalopram, and three metabolites (norfluoxetine, demethylcitalopram and didemethylcitalopram), on S-mephenytoin 4' hydroxylation activities in human liver microsomes were studied. The 4' hydroxylation of S-mephenytoin, a representative substrate toward CYP2C19, was competitively inhibited by all the SSRIs and their metabolites studied. The mean Ki values of fluoxetine, norfluoxetine, sertraline, paroxetine, citalopram, demethylcitalopram and didemethylcitalopram were 5.2, 1.1, 2.0, 7.5, 87.3, 55.8 and 7.7 microM, respectively. The findings suggest that some SSRIs and their metabolites with a low Ki value (e.g., fluoxetine, norfluoxetine) may reduce the clearance of drugs metabolized by this isoform of P450, thereby resulting in a possible drug-drug interaction, when administered simultaneously. In addition, SSRIs and their metabolites examined herein may be substrates toward CYP2C19. PMID- 8703654 TI - The effect of meloxicam on the pharmacokinetics of beta-acetyl-digoxin. AB - The influence of multiple-dose administration of meloxicam on the pharmacokinetics of oral beta-acetyl-digoxin was studied in 12 healthy male volunteers in a randomized double-blind two-way crossover study. The primary endpoint, Cminss, was within the accepted range for bioequivalence, as were Cmaxss and AUCss. The 90% confidence interval and the point estimator of 98.7 for Cminss were within the equivalence range of 0.8-1.25. MRT and tmax were also unchanged, while the elimination rate constant was decreased slightly by 12%, which is of no therapeutic relevance. It is concluded that co-treatment with meloxicam has no effect on the pharmacokinetics of oral digoxin. PMID- 8703655 TI - Ethnic differences in nifedipine kinetics: comparisons between Nigerians, Caucasians and South Asians. AB - Nifedipine was administered to 12 healthy Nigerian volunteers as a single oral dose of 20 mg capsule under fasting conditions. The pharmacokinetic results were compared with published data using the same protocol and analytical method for 27 Caucasians and 30 South Asians. The area under the plasma concentration-time curve (AUC) of nifedipine in Nigerians (808 +/- 250 ng ml-1 h) was significantly higher (P < 0.001) than that in Caucasians (323 +/- 116 ng ml-1 h) and the difference remained significant (P < 0.001) when corrected for body weight. The elimination half-life was also significantly higher (P < 0.01) in Nigerians (5.03 +/- 1.96 h) than in Caucasians (2.78 +/- 1.11 h). No significant differences were observed between Nigerians and South Asians in either AUC or half-life of nifedipine. The AUC of the nitropyridine metabolite was higher (P < 0.01) in Nigerians (220 +/- 51 ng ml-1 h) compared with that in Caucasians (154 +/- 56 ng ml-1 h) but the difference was not maintained when corrected for body weight. The AUC corrected for body weight and the elimination half-life of the metabolite were significantly higher in South Asians compared with those of Nigerians and Caucasians. The pharmacokinetics of oral nifedipine in Nigerians were similar to those in South Asians and therefore may also arise from a lower systemic clearance compared with Caucasians as has been reported previously for South Asians. PMID- 8703656 TI - Pharmacokinetics of quinine in chronic liver disease. AB - The pharmacokinetics of quinine were investigated in a) six healthy male Thai subjects, and b) nine male Thai patients with a moderate degree of chronic liver disease, after a single oral dose of 600 mg quinine sulphate. tmax and t1/2.2 were significantly prolonged in patients (median [range] tmax 2 [1-5] vs 1.6 [0.8 2] h; t1/2,z 23.4 [17.4-41.7] vs 9.7 [7.8-17.2] h), and Vz/F was significantly larger (median [range] 4.21 [2.33-15.87] vs 2.78 [1.49-3.38] 1 kg-1). Median (range) concentration of the plasma unbound Qn fraction collected from the patients at 4 h after drug administration was 17 (8.4-17.8)% of total drug concentration. PMID- 8703657 TI - The role of human cytochrome P450 enzymes in the metabolism of anticancer agents: implications for drug interactions. AB - 1. Little information is available about the pharmacokinetic interactions of anticancer drugs in man. However, clinically significant drug interactions do occur in cancer chemotherapy, and it is likely that important interactions have not been recognized. 2. Specific cytochrome P450 (CYP) enzymes have been recently shown to be involved in the metabolism of several essential anticancer agents. In particular, enzymes of the CYP3A subfamily play a role in the metabolism of many anticancer drugs, including epipodophyllotoxins, ifosphamide, tamoxifen, taxol and vinca alkaloids. CYP3A4 has been shown to catalyse the activation of the prodrug ifosphamide, raising the possibility that ifosphamide could be activated in tumour tissues containing this enzyme. 3. As examples of recently found, clinically significant interactions, cyclosporin considerably increases plasma doxorubicin and etoposide concentrations. Although cyclosporin and calcium channel blockers may influence the pharmacokinetics of certain anticancer agents by inhibiting their CYP3A mediated metabolism, it is more likely that these P glycoprotein inhibitors inhibit P-glycoprotein mediated drug elimination. 4. Appropriate caution should be exercised when combining P-glycoprotein inhibitors and potential CYP3A inhibitors with cancer chemotherapy. PMID- 8703658 TI - N-Hydroxylation of dapsone by multiple enzymes of cytochrome P450: implications for inhibition of haemotoxicity. AB - 1. The adverse reactions associated with the administration of dapsone are believed to be caused by metabolism to its hydroxylamine. Previous reports suggest that CYP3A4 is responsible for this biotransformation [1]. 2. Data presented in this paper illustrate the involvement of more than one cytochrome P450 enzyme in dapsone hydroxylamine formation using human liver microsomes. Eadie-Hofstee plots demonstrated bi-phasic kinetics in several livers. No correlation could be established between hydroxylamine formation and CYP3A concentrations in six human livers (r = -0.47; P = 0.34). 3. Studies with low molecular weight inhibitors illustrate the importance of CYP2C9 and CYP3A in dapsone N-hydroxylation. 4. Differential sensitivity of dapsone N-hydroxylation to selective CYP inhibitors indicated that the contribution of individual CYP enzymes varies between livers. Selective inhibition ranged from 6.8 to 44.1% by 5 microM ketoconazole, and from 24.0 to 68.4% by 100 microM sulphaphenazole. The extent of inhibition, by either ketoconazole or sulphaphenazole was dependent on the CYP3A content of the liver. 5. The levels of expression of these cytochrome P450 enzymes may be an important determinant of individual susceptibility to the toxic effects of dapsone, and may influence the ability of an enzyme inhibitor to block dapsone toxicity in vivo. Because of the inability to produce complete inhibition, selective CYP inhibitors are unlikely to offer any clinical advantage over cimetidine in decreasing dapsone hydroxylamine formation in vivo. PMID- 8703659 TI - Dose-response relationship and time-course of the effect of inhaled magnesium sulphate on airflow in normal and asthmatic subjects. AB - 1. Magnesium is a dietary cation with a wide range of actions of potential relevance to asthma. 2. To determine the dose-response relationship and time course of the effect of inhaled magnesium sulphate on the airway, we have studied the effect of 0, 90, 135, 180 and 360 mg of magnesium sulphate given by nebulizer on specific airways conductance (sGaw) in 20 normal subjects, and forced expiratory volume in one second (FEV1), forced vital capacity (FVC), flow at 25% forced vital capacity (Vmax25) and peak expiratory flow (PEF) in 19 asthmatic subjects. 3. On five occasions after baseline measurements of airway calibre, one of the five doses of magnesium sulphate in 3 ml normal saline was administered by nebulizer in a randomized, double-blind design. Measurements of sGaw or FEV1, FVC, Vmax25 and PEF were made at 5 and 10 min after nebulization and at 10 min intervals thereafter up to 90 min. 4. There was no significant difference in the mean area under the curve (AUC) for change from baseline in sGaw or maximum increase from baseline between doses in normal subjects. 5. In asthmatic subjects there was no significant difference in the mean AUC for change from baseline in FEV1, FVC or Vmax25 when compared between doses by analysis of variance. There was a difference in the mean AUC for change from baseline in PEF between doses (ANOVA P for all groups 0.052) but this can be explained by a detrimental effect of the maximum dose of magnesium sulphate. 6. It would appear that inhaled magnesium does not act as a bronchodilator in normal or asthmatic subjects. PMID- 8703660 TI - Effects of azapropazone on pain-related brain activity in human subjects. AB - 1. The dose-related effects of azapropazone on (i) event-related and spontaneous EEG-activity and (ii) the subjects' pain ratings were investigated using an experimental human pain model based on both chemo-somatosensory event-related potentials (CSSERP) and subjects' pain ratings. 2. Healthy subjects (n = 20) participated in a placebo-controlled, randomized, double-blind, four-way cross over study. Single doses of azapropazone (300 mg, 600 mg and 1200 mg) and placebo were administered intravenously. Each experiment consisted of five sessions (before and 1, 2, 4 and 8 h after administration of the medication). Each session lasted for approximately 40 min. In the first 20 min, pain was induced by short CO2-stimuli presented to the right nostril (phasic pain; interstimulus interval 30 s) and EEG was recorded from five positions. CSSERPs were obtained in response to painful CO2-stimuli. In the following 20 min period, tonic pain was induced by a constant stream of dry air introduced in the left nostril. Subjects rated the intensity of both phasic and tonic pain by means of a visual analogue scale. Additionally, a frequency analysis of the spontaneous EEG was performed. 3. Azapropazone reduced the pain-related CSSERP-amplitudes at frontal and parietal recording positions. This topographical pattern was observed in previous studies with opioids, while NSAIDs such as flurbiprofen and ketoprofen exerted effects at frontal and central positions. In contrast to other NSAIDs, administration of azapropazone resulted in a reduction of the frequency bands alpha 1, delta and theta of the spontaneous EEG. At the subjective level, analgesic effects of azapropazone were observed in the ratings of tonic pain. 4. Analgesic properties of azapropazone were demonstrated in man. The topographical pattern of the changes in the CSSERPs and the effects on EEG background activity suggest a central component of the analgesic action of azapropazone. PMID- 8703661 TI - Plasma concentrations of fentanyl with subcutaneous infusion in palliative care patients. AB - 1. Plasma concentrations of fentanyl were measured by g.c. in 20 patients (median age: 75 years and range: 54-86 years; eight females) in palliative care receiving the drug by continuous s.c. infusion (median rate: 1200 micrograms day-1 and range: 100-5000 micrograms day-1). 2. The infusion rate was significantly related to the duration of therapy (Spearman rho = 0.56, P < 0.05). The total steady state plasma concentrations of fentanyl ranged between 0.1 and 9 ng ml-1, with a median of 1 ng ml-1. The unbound fraction of fentanyl in the plasma ranged from 17.8 to 44.4%, with a median value of 33.6%. Infusion rates and both total and unbound plasma concentrations of fentanyl were correlated (Spearman rho = 0.92, P < 0.05 in each case). Even with standardization for dosage, there was an eightfold variation in total plasma concentrations and 3.5-fold variation in unbound plasma concentrations of fentanyl. 3. There is considerable inter-patient variability in the pharmacokinetics of fentanyl with s.c. infusion in the palliative care setting, which necessitates careful titration of dosage according to individual clinical response. PMID- 8703662 TI - Actions of terfenadine and cimetidine on histamine wheal formation. AB - 1. A Latin square design was used to compare the effects of four masked oral drugs, placebo, cimetidine, terfenadine and cimetidine plus terfenadine, at single standard doses against histamine injected at concentrations of 0.3 mM and 0.074 mM subepidermally in eight healthy volunteers studied at weekly intervals. 2. Wheal growth occurred in three phases, an initial delay, a phase of rapid growth where drugs were effective, and a phase of slow growth where there were no significant drug effects. 3. Terfenadine was an effective antihistamine. 4. The data were consistent with a terfenadine effect continued into the next treatment block. 5. Under these conditions cimetidine apparently reduced the effects of terfenadine during the rapid wheal growth phase both simultaneously and after a week's interval from terfenadine dosing. PMID- 8703663 TI - The effects of rilmenidine and atenolol on mental stress, dynamic exercise and autonomic function in mild to moderate hypertension. AB - 1. The effects of 4 week treatment with rilmenidine or atenolol on tests of mental stress, dynamic exercise, autonomic function and psychometric tests were evaluated in a randomized, double-blind, placebo-controlled, cross-over study. 2. After a 4 week placebo run-in, 12 patients with essential hypertension (blood pressure [BP] 160/95 +/- 15/7 mmHg) received rilmenidine 1-2 mg day-1, and atenolol 50-100 mg day-1, each for 4 weeks, with a 4 week placebo wash-out between drug treatments. 3. Both agents produced a comparable reduction in supine and erect BP. During the mental arithmetic test, BP and heart rate (HR) responses were similar for rilmenidine and atenolol. 4. During bicycle exercise, the increase in HR was significantly greater after rilmenidine (+50 vs 41 beats min 1, P = 0.04). During recovery, the areas under the curve for diastolic BP (46,450 vs 51,400 mmHg s, P = 0.02) and HR (49,445 vs 63,597 beats min-1 s, P = 0.001) were significantly less with atenolol than rilmenidine. 5. Neither rilmenidine nor atenolol affected mental performance as judged by arithmetic and psychomotor tests. Physiological responses to autonomic function tests (deep breathing, facial immersion, isometric handgrip and cold pressor) were preserved with both drugs. The standing to lying ratio was higher on atenolol (P = 0.01) and Valsalva ratio was higher on rilmenidine (P = 0.03). 6. In conclusion, rilmenidine and atenolol exerted comparable antihypertensive effects both at rest and during mental and dynamic stress. Atenolol attenuated HR responses to dynamic exercise and the Valsalva manoeuvre; rilmenidine did not interfere with the physiological responses of BP and HR during autonomic function tests. PMID- 8703664 TI - Effect of multiple doses of losartan on the pharmacokinetics of single doses of digoxin in healthy volunteers. AB - 1. Losartan (DuP 753, MK-954) is a novel, potent and highly selective AT1 angiotensin II receptor antagonist. The effect of multiple oral doses of losartan on digoxin pharmacokinetics was evaluated in healthy male subjects. 2. In a double-blind and randomized fashion, subjects received 50 mg losartan or placebo once daily for 15 days in each period. At least 7 days elapsed between the two treatment periods. On days 4 and 11 of each period, subjects also received a single 0.5 mg dose of digoxin intravenously and orally respectively. 3. Eleven of 13 subjects completed the study. Side effects were mild and transient (12 out of 13 subjects reported at least one adverse experience). During the study, no laboratory abnormalities were noted. 4. Multiple oral doses of losartan (50 mg daily) did not affect the pharmacokinetic parameters of 0.5 mg of digoxin i.v. AUC(0.48h) of immunoreactive digoxin during losartan 28.8 +/- 2.9 vs 28.5 +/- 3.9 ng ml-1 h during placebo; not significant, and 96 h urinary excretion [% dose] during losartan 54.0 +/- 7.2 vs 51.9 +/- 6.5% during placebo; not significant). Geometric mean ratios (90% confidence interval) for AUC and urinary excretion were respectively, 1.03 (0.98, 1.08) and 1.09 (0.98, 1.21). 5. Multiple oral doses of losartan did not affect the pharmacokinetic parameters of oral digoxin AUC(0.48 h) during losartan 23.6 +/- 3.7 ng ml-1 h vs 22.4 +/- 2.6 ng ml-1 h during placebo; not significant, Cmax 3.5 +/- 0.7 ng ml-1 with vs 3.1 +/- 0.5 ng ml-1 without losartan; not significant and tmax 0.6 +/- 0.2 h with vs 0.9 +/- 0.7 h without losartan; not significant, and 96 h urinary excretion [% dose] during losartan 51.2 +/- 6.3 vs 46.3 +/- 2.4% during placebo; not significant). Geometric mean ratios (90% confidence interval) for AUC and urinary excretion were respectively, 1.06 (0.98, 1.14) and 1.12 (0.97, 1.28). 6. We conclude that multiple oral doses of losartan (50 mg daily) do not alter the pharmacokinetics of immunoreactive digoxin, following either intravenous or oral digoxin. Furthermore, the co-administration of digoxin with losartan is well tolerated by healthy male volunteers. PMID- 8703665 TI - Comparison of biological activities of two low molecular weight heparins in 10 healthy volunteers. AB - 1. Low molecular weight heparins (LMWHs) are produced by different depolymerization processes and may therefore differ with respect to their pharmacokinetic properties. 2. We designed a single dose, randomized cross-over study in 10 healthy volunteers to compare the 24 h pharmacokinetics of two LMWHs, reviparin and enoxaparin, which have been previously shown to be clinically equivalent in terms of post-operative deep vein thrombosis prevention, despite significant differences in their in vivo biological activity. The two LMWHs were subcutaneously administered at the same dosages that are used in clinical studies: 4250 anti-Xa iu for reviparin and 40 mg for enoxaparin which have similar in vitro anti-Xa activities. 3. The overall 24 h profiles of the plasma anti-Xa and anti-thrombin activities were similar for reviparin and enoxaparin. The Amax and the AUC(0, 24h) of plasma anti-Xa activity after reviparin administration were both slightly but significantly lower than those observed after enoxaparin administration (difference between treatments of 0.03 95% CI[0.01-0.05] iu ml-1 h and 0.56 95% CI[0.22-0.90] iu ml-1 for Amax and AUC(0, 24h) respectively). After adjustment for in vitro anti-Xa activity, the statistical difference between the two LMWHs persisted for the AUC(0, 24h) but not for the Amax of plasma anti-Xa activity. The tmax and the MRT values for plasma anti-Xa activity did not significantly differ between the two drugs. The t1/2 for reviparin did not significantly differ from that of enoxaparin (2.7 +/- 0.7 h vs 3.5 +/- 0.9 h respectively, NS). The Amax of the plasma anti-thrombin activity after reviparin administration was also slightly but significantly lower than that observed after enoxaparin administration, (difference between treatments of 0.018 95% CI[0.01-0.025] iu ml-1) whereas the AUC(0, 24h) of anti thrombin activity vs time was not. A slight but significant increase of the activated partial thromboplastin time of a similar magnitude was observed after both reviparin and enoxaparin injections. 4. The calculated surface under the thrombin generation curve vs time (or thrombin potential) at peak was significantly higher after reviparin than after enoxaparin (367 +/- 53 UA vs 305 +/- 48 UA respectively, P < 0.05). Four hours after injection, thrombin potential was significantly correlated to plasma anti-Xa activity after reviparin but not after enoxaparin injection (r = 0.65, n = 10, P = 0.05 and r = -0.38, n = 10, P = 0.25 respectively). 5. After a single-dose injection in healthy subjects, two LMWHs with comparable in vitro activities differed slightly kinetically. Such minor differences are probably of little importance in the prevention of post operative deep vein thrombosis, since these two LMWHs were previously shown to be comparable in this setting. PMID- 8703666 TI - Acute effects of ANP and BNP on hypoxic pulmonary vasoconstriction in humans. AB - 1. Atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) have pulmonary vasorelaxant activity with plasma concentrations being elevated in patients with hypoxaemic pulmonary hypertension. However, their effects on acute hypoxic pulmonary vasoconstriction (HPV), the initiating stimulus for pulmonary hypertension have not to date been investigated. We have therefore studied the effects of ANP and BNP on acute HPV in humans. 2. Eight healthy volunteers were studied on three separate occasions. After reaching a resting haemodynamic state (t0), an infusion of either ANP (10 pmol kg-1 min-1), BNP (10 pmol kg-1 min-1) or placebo (5% dextrose) was commenced. This was given alone for 30 min (t30) before subjects were rendered hypoxaemic (SaO2 75-80%) for a further 30 min (t60), with the initial infusion continuing to t60. Pulsed-wave Doppler analysis of pulmonary artery flow was used to measure mean pulmonary arterial pressure (MPAP) and hence total pulmonary vascular resistance (PVR) was calculated. 3. MPAP and PVR both tended to decrease in response to ANP and BNP infusion, although compared with placebo, the difference at t30 was only statistically significant for PVR. Hypoxaemia increased MPAP and PVR, although values at t60 were significantly lower following both ANP and BNP compared with placebo. 4. In terms of the actual change in PVR (delta PVR) induced by hypoxaemia (from t30 to t60), BNP (146(16) dyn s cm-5), but not ANP (183(21) dyn s cm-5) significantly attenuated delta PVR compared with placebo (194(26) dyns s cm-5): mean difference BNP versus placebo 48 dyn s cm-5, 95% Cl 3-93. An identical pattern was observed for delta MPAP where BNP (15.9(1.1) mmHg), but not ANP (18.0(1.2) mmHg) significantly attenuated delta MPAP compared with placebo (19.0(1.7) mmHg): mean difference BNP versus placebo 3.1 mmHg, 95% Cl 0.7-5.5. 5. Thus, although both ANP and BNP exhibit pulmonary vasorelaxant activity, only BNP significantly attenuated the MPAP and PVR responses to acute hypoxaemia. This suggests that the natriuretic peptides may have a role in attenuating pulmonary hypertension secondary to hypoxaemia. PMID- 8703667 TI - The effect of HN-65021 on responses to angiotensin II in human forearm vasculature. AB - We studied the effect of (2-butyl-4-chloro-1[[2'-(1H-tetrazol-5-yl) [1,1' biphenyl]methyl]-1H-imadazole-5-carboxylic acid,-1-(ethoxycarbonyloxy) ethyl ester (HN-65021), on angiotensin II induced vasoconstriction in forearm vasculature of eight healthy men. Placebo and HN-65021 (5, 10 and 100 mg) were administered orally. Forearm blood flow was measured by venous occlusion plethysmography during rising dose brachial artery infusions of angiotensin II (0.3-1000 pmol min-1) 2 h after dosing. HN-65021 inhibited angiotensin II, causing a shift to the right of the dose-response curve. Angiotensin II (100 pmol min-1) decreased mean blood flow in the infused arm by 63.1 +/- 3.2% when infused following placebo and by 49.9 +/- 4.3%, 50.7 +/- 3.5% and 36.4 +/- 2.8% following HN-65021 doses of 5.10 and 100 mg respectively. These results demonstrate that HN 65021 antagonises angiotensin II receptor mediated vasoconstriction in human forearm resistance vessels. PMID- 8703668 TI - Binding of temoporfin to the lipoprotein fractions of human serum. AB - The binding of a new photosensitizer, temoporfin, to human serum lipoproteins was investigated. [14C]-Temoporfin (0.1-10 micrograms ml-1) was incubated with human serum for 30 min at room temperature or for 20 h at 4 degrees C, prior to stepwise density flotation to separate the lipoprotein fractions. The distribution of the drug was independent of the initial concentration or time and temperature of the incubation. The proportion of temoporfin in each fraction was: very low density lipoprotein 6%, low density lipoprotein 22%, lipoprotein(a) 17%, high density lipoprotein 39% and lipoprotein deficient serum 16%. Autoradiography of agarose gels showed that the drug was associated with the lipoprotein in the fractions. Fractionation of plasma samples collected from a patient after an intravenous infusion of temoporfin revealed a binding profile similar to that obtained in the in vitro study. PMID- 8703669 TI - Lack of an effect of nefazodone on the pharmacokinetics and pharmacodynamics of theophylline during concurrent administration in patients with chronic obstructive pulmonary disease. AB - The effect of nefazodone on the pharmacokinetics and pharmacodynamics of theophylline was evaluated in a multiple-dose, randomized placebo-controlled, double-blind two-period crossover study in 13 patients who were undergoing theophylline therapy for chronic obstructive pulmonary disease. Two treatments were administered, each for 7 days: theophylline + 200 mg nefazodone twice daily (every 12h) and theophylline+matching nefazodone placebo capsule twice daily (every 12h). Mean peak and trough plasma concentrations of theophylline ranged from 13.1 to 14.5 micrograms ml-1 and 11.6 to 14.2 micrograms ml-1, respectively, at steady-state when theophylline was administered with or without concurrent dosing of nefazodone. Similarly, the mean area under the curve for theophylline ranged from 93.5 to 103 micrograms ml-1 h. When nefazodone and theophylline were co-administered, theophylline pharmacokinetic parameters did not significantly differ from those obtained when theophylline was administered with placebo. Forced expiratory volume in one second (FEV1) measurements taken when nefazodone or placebo were administered with theophylline did not differ from those obtained at baseline. The plasma concentration-time profiles for nefazodone and its metabolites were similar to those in other studies where nefazodone was administered alone. Since nefazodone did not affect the pharmacokinetics or the pharmacodynamics of theophylline, no change in theophylline dose should be needed as a consequence of nefazodone co-administration. PMID- 8703670 TI - Scientific activities of Professor Lindsay Symon in the field of clinical neurosurgery. PMID- 8703671 TI - The experimental writings of Lindsay Symon. PMID- 8703672 TI - The ischaemic penumbra: twenty years on. AB - In focal ischaemia, the penumbra defines regions with blood flow below that needed to sustain electrical activity, but above that required to maintain cellular ionic gradients, and that lead in time to infarction. Among other terms used to describe regions surrounding the ischaemic core, "penumbra" is the only one based on a precise functional state of partially ischaemic tissue. The precarious balance between energy supply and demand that characterizes the penumbra and the proximity of the ischaemic core are the basis of a number of important features: (a) It is a time-limited condition, with a tendency to evolve towards infarction and to propagate to adjacent viable tissue; (b) "misery perfusion" is associated with increased oxygen extraction, acidosis, and high glucose utilization, but residual ATP; (c) recurrent spreading depression contributes to the deterioration of the penumbra, at least in animal models of stroke; (d) there is no sustained increase of extracellular glutamate; and (e) improvement of local perfusion and reduction of energy demand remain the most rational approaches to rescue the penumbra. By defining a window of opportunity for therapeutic intervention in stroke, the concept of ischaemic penumbra has enormously stimulated research in this field and led to a better understanding of the pathophysiology of cerebral ischaemia, with direct practical application for the surgical management of cerebrovascular disorders such as aneurysms. PMID- 8703674 TI - Neurogenic control of the cerebral circulation. PMID- 8703673 TI - Nitric oxide and cerebral blood flow: an update. AB - This article focuses on the key concept that a basal production of nitric oxide (NO) is required as a background for biological modulation, although an excess can be cytotoxic. Studies of ischaemia and neurodegeneration have tended to emphasise detrimental effects of excess NO, but this review contrasts the emerging importance of diminished NO or interference with its action in vasospasm following subarachnoid haemorrhage (SAH) in ageing and in atherosclerosis. Clinical intervention in cerebral ischaemia will require specificity of action, since NO appears to be protective or detrimental depending on the time, source, and distribution of its production. It may be possible to utilise targeted action on the different forms of NO synthase or the specific redox forms of NO in different tissue areas. PMID- 8703675 TI - AUA's response to the OTA study on prostate cancer screening for elderly men. PMID- 8703676 TI - Physicians urged to address quality of life after breast cancer treatment. PMID- 8703677 TI - Initiatives can contain costs and increase quality of nursing care. PMID- 8703678 TI - Networking may be key to academic cancer center survival under managed care. PMID- 8703679 TI - Many patients said to leave their MD's office without understanding what they've been told. PMID- 8703680 TI - ACCC clinical investigators cite negative impact of managed care and cost containment on oncology clinical trials. PMID- 8703681 TI - Chances of worldwide plagues increasing, scientists say. PMID- 8703682 TI - Infectious disease testing for blood transfusions. PMID- 8703683 TI - New chemoradiation approach for bladder cancer tested in RTOG 95-06. PMID- 8703684 TI - Systemic treatments for advanced cutaneous melanoma. AB - The treatment of advanced cutaneous melanoma remains disappointing. Single-agent cytotoxic drugs usually produce response rates of less than 20%, though newer agents, particularly fotemustine and temozolomide, show some promise, especially in patients with brain metastases. Combination chemotherapy regimens yield response rates of 20% to 40%, but durable complete remissions are uncommon. Interferon-alfa and interleukin-2 alone produce response rates of 10% to 20%, 3% to 5% of which are durable. Vaccines and monoclonal antibodies have low level activity in advanced disease but may play a role in the adjuvant setting. The combinations of cisplatin-based regimens plus IFN-alfa and IL-2 have produced overall response rates of 50% to 60% and complete responses in 20% of patients, about half of which are durable. The toxicity of these regimens is severe, however, and their impact on survival remains to be established. PMID- 8703686 TI - Chemotherapy enhancer passes first test. PMID- 8703685 TI - Clinical trials. Referral resource. Clinical trials assessing quality of life. PMID- 8703687 TI - Ribozyme gene therapy for cancer nears clinical trials. PMID- 8703688 TI - New anticancer agents in clinical development. AB - A better understanding of the biology and biochemistry of the cancer cell has led to the development of various promising new antineoplastic compounds that are now undergoing phase I, II, and III clinical testing. These drugs include topoisomerase I inhibitors, such as camptothecin and its analogs 9 aminocamptothecin, irinotecan, and topotecan; the paclitaxel analog docetaxel; gemcitabine, an antimetabolite structurally related to cytarabine; and fluorouracil prodrugs and other thymidylate synthase (TS) inhibitors. Another exciting approach to cancer treatment is the use of agents that induce a less malignant state by altering cellular phenotype. Such agents include angiogenesis inhibitors, differentiating agents, signal transduction inhibitors, and gene therapy. PMID- 8703689 TI - PET scanning said to save money and improve staging of lung cancer. PMID- 8703690 TI - An overview of radiotherapy trials for the treatment of brain metastases. AB - A review of the English literature was undertaken to (1) determine the efficacy of radiation therapy for the treatment of brain metastases, (2) identify prognostic factors, and (3) ascertain whether there is an effect of treatment technique on outcome. Critical analysis of relevant randomized trials indicated that radiation therapy can effectively palliate the symptoms of brain metastases. Prognostic factors for improved survival are good performance status and the absence of a non-central nervous system tumor. The most efficient treatment protocol is controversial, but the literature supports the use of 20 Gy in five fractions for the treatment of patients with a poor prognosis. Patients with a solitary brain metastasis and no systemic disease benefit from resection of the brain metastasis followed by postoperative radiation. PMID- 8703691 TI - Changes in laryngeal preservation study help increase accrual. PMID- 8703692 TI - New 'calibrator' should improve prostate cancer testing. PMID- 8703693 TI - Current techniques in three-dimensional CT simulation and radiation treatment planning. AB - The modern CT simulator is capable of interactive three-dimensional (3D) volumetric treatment planning; this allows radiation oncology departments to operate without conventional X-ray simulators. Treatment planning is performed at the time of virtual simulation by contouring the organs or volumes of interest and determining the isocenter. A digitally reconstructed radiograph (DRR) provides a beam's-eye-view display of the treatment field anatomy and contoured areas of interest. Conformal and noncoplanar teletherapy is facilitated for patients with prostate cancer, lung cancer, and brain tumors. Ongoing developments include 3D dose calculation, dose-volume histogram analysis, and tumor dose escalation. PMID- 8703695 TI - Are there restrictions to vaccinating a child who has recovered from hemolytic uremic syndrome? PMID- 8703694 TI - Congenital nephrotic syndrome of the Finnish type: linkage to the locus in a non Finnish population. AB - Congenital nephrotic syndrome of the Finnish type (CNF) is inherited as an autosomal recessive trait. The biochemical basis of the disease is unknown, although a lesion in the glomerular basement membrane is strongly suggested. Recently, the CNF locus was assigned to chromosome 19q12-q13.1 on the basis of linkage analysis in Finnish families. The high incidence of the disease in Finland, as well as the demonstration of linkage disequilibrium in the Finnish study, strongly suggests a founder effect based on a common ancient mutation in this population. We confirm linkage of the CNF locus to the same chromosomal region in seven non-Finnish CNF families without evidence of linkage disequilibrium. Our results show that the same gene seems to be affected in both Finnish and non-Finnish CNF populations. However, in the latter the mutation carrying chromosomes descend from different ancestors without evidence of a founder effect. PMID- 8703696 TI - Girls prone to urinary infections followed into adulthood. Indices of renal disease. AB - This study describes blood pressure and renal function, as well as indices of renal disease, in females with and without renal scarring followed from their first urinary tract infection (UTI) in childhood. Of the 111 patients with a median follow-up time of 15 years, 54 had renal scarring (reflux nephropathy) on urography, which was severe in 19 and moderate in 35. The glomerular filtration rate was lower in patients with severe renal scarring and correlated with renal area on urography. However, the filtration rate was decreased below the lower reference limit in only 7 patients, with a lowest value of 70 ml/min per 1.73 m2. The diastolic blood pressure was higher in women with severe scarring. Hypertension of at least 140/90 mmHg was diagnosed in 3 of 54 (5.5%) females with renal scarring, 2 before and 1 at the follow-up examination. The excretion of albumin in urine was low and not correlated to filtration rate. Tubular enzymes in urine were similar in all groups. Thus the renal function was well preserved and the incidence of hypertension low. Within this range of renal function, the level of albumin in urine did not predict the degree of renal scarring. PMID- 8703697 TI - Posterior urethral valves in patients with Down syndrome. AB - Renal and urological anomalies in Down syndrome (DS) have received little attention compared with the nephrourological findings described in other chromosomal abnormalities. Renal hypoplasia, hydroureteronephrosis, ureterovesical and ureteropelvic junction obstruction, and vesicoureteral reflux, but not posterior urethral valves, have been associated with DS. We report the occurrence of posterior urethral valves in three male infants with DS at a single institution. All had multiple urological procedures for correction or palliation of obstruction. Children with DS may have an increased risk for developing posterior urethral valves and obstructive uropathy. Furthermore, they may also develop chronic renal failure secondary to posterior urethral valves. Therefore, we suggests that infants with DS be screened with ultrasonography for renal and urological abnormalities early in life and, if abnormal, a contrast voiding cystourethrogram be performed to rule out posterior urethral valves or other bladder or urethral abnormalities. A review of the renal and urological anomalies in DS reported in the literature since 1960 is presented. PMID- 8703698 TI - What is the actual management of a child with membranous glomerulopathy associated with chronic hepatitis B? PMID- 8703699 TI - Renal artery stenosis in infants: long-term medical treatment before surgery. AB - This report describes five infants (3 male, 2 female) with renal artery stenosis diagnosed in their 1st year of life. The age at initial presentation was 5 days to 10 months. All had symptoms of congestive heart failure, cardiomegaly on chest X-ray, and left ventricular hypertrophy by electrocardiogram or echocardiogram. Renograms were abnormal in four of the five infants. An intravenous pyelogram was obtained in three infants and was abnormal in two. Renal ultrasounds were obtained in two infants and were normal in both. Patients were treated for 4.4 +/ 0.9 years with antihypertensive drug therapy until surgical correction of the renal artery stenosis. Blood pressure was persistently elevated above the 95th percentile in four of the infants during the course of antihypertensive therapy prior to surgery. Patients have been followed for 9.4 +/- 2 years since surgery. The blood pressure of four patients is normal, and the blood pressure of the oldest patient (age 23 years) is borderline hypertensive. These data show that infants with renal artery stenosis can be cared for successfully with long-term antihypertensive drug therapy to preserve renal mass with minimal chronic adverse effects. PMID- 8703700 TI - Rate of change of blood pressure in premature and full term infants from birth to 4 months. AB - Systolic and diastolic blood pressures were evaluated in a cohort of 61 non hypertensive premature [very low birth weight (VLBW), n = 16; low birth weight (LBW), n = 22] and full-term [normal birth weight (NBW), n = 23] newborn infants admitted to a neonatal intensive care unit (NICU) and followed to their 4-month age-adjusted outpatient examination. All were receiving routine postnatal care by 7 days of age. Blood pressure was measured at 7 days of age, at discharge from the NICU, and at the outpatient examination. Simple linear regression of blood pressure on weight was used to fit a straight line to the three measurements for each infant and the average regression line for each birth weight group was then obtained. There was a significant correlation between systolic blood pressure and both weight and length at each of the measurement points and also between the change in systolic blood pressure and change in weight from the discharge to the 4-month examination. Diastolic blood pressure tended to follow this same pattern. Gestational age was correlated significantly with the 7-day blood pressure, but postnatal age at the outpatient examination was not correlated with either systolic or diastolic blood pressure. The average slopes of systolic and diastolic blood pressure on weight (mmHg/kg body weight) were virtually identical for the LBW and NBW groups; in contrast, the average slope of the VLBW group was greater than the other two groups, and the difference was statistically significant for diastolic blood pressure. These results show significant group differences in mean blood pressure prior to 4 months of age between VLBW, LBW, and NBW groups and, for the VLBW infants, a steeper slope of the estimated regression line of blood pressure on weight between birth and 4 months. PMID- 8703701 TI - Doppler studies in normal kidneys of healthy children. AB - Despite the widespread use of Doppler ultrasound in kidney diseases of adults and children, there are only a few, partially contradictory reports of normal values of resistance index (RI) and pulsatility index (PI) of renal perfusion in a healthy pediatric population. This study was carried out to determine normal values for these parameters in a larger group of probands. For the RI correlation with age was most pronounced in the right renal artery, with children over 6 years having a significantly lower RI than younger children. The PI was relatively independent of side and age. These results are somewhat different to previous reports. These differences may influence future Doppler sonographic investigations of acute and symmetrically affected kidneys (nephrological diseases). Our data suggest that the RI of the right renal artery might be most useful. In chronical or unilateral kidney diseases (unilateral ureteral obstruction, reflux, or operation) investigation of the PI in addition to the RI may aid diagnosis. PMID- 8703702 TI - Family behavior, adaptation, and treatment adherence of pediatric nephrology patients. AB - In this exploratory study we investigated the relationships among family behavior variables (e.g., family expressiveness), adaptive functioning skills, maladaptive behavior, and adherence to treatment in pediatric renal failure patients. The study included 22 pediatric outpatients with renal failure who had not yet received dialysis or transplantation (RF) and their parents, and 12 pediatric outpatients with kidney transplants (TX) and their parents. For the RF patients, significant correlations were found between some of their adaptive functioning skills and measures of their medication adherence, diet adherence, and clinic appointment adherence; however, for the TX patients significant correlations were found only between some of their adaptive functioning skills and measures of their medication adherence. For the RF patients only, some measures of their family behavior were significantly correlated with measures of their medication adherence and diet adherence. Additionally, some measures of the RF patients' family behavior were significantly related to their communication skills, socialization skills, overall adaptive functioning skills, and maladaptive behavior. For the TX patients, only their socialization skill level was significantly correlated with one measure of their family behavior. It is concluded that facilitation of adaptive and physical functioning among renal pediatric patients likely requires multidimensional training and/or counselling interventions with the children and their families, and that some of the content and/or emphasis of this training likely needs to differ for RF patients versus TX patients. PMID- 8703703 TI - Poor renal uptake of 99mtechnetium-dimercaptosuccinic acid and near-normal 99mtechnetium-mercaptoacetyltriglycine renogram in nephronophthisis. AB - Four patients with the clinical diagnosis of nephronophthisis are presented, all having a very poor renal uptake of 99mtechnetium-dimercaptosuccinic acid (99mTc DMSA) but clearly visualized kidneys on early images with 99mtechnetium mercaptoacetyltriglycine and a normal or almost normal renogram. There was no difference between a young patient in an early stage of the disease and the other three patients with more advanced renal disease. In contrast, a patient with tubulointerstitial nephritis with uveitis had considerably better renal uptake of 99mTc-DMSA despite impaired renal function. We suggest that the specific tubular function defect in nephronophthisis might be the cause of the poor uptake of 99mTc-DMSA. We also recommend the method to support the clinical suspicion of nephronophthisis, even in the early stages of the disease. PMID- 8703704 TI - Hyperlipidaemia, diet and simvastatin therapy in steroid-resistant nephrotic syndrome of childhood. AB - In children with steroid-resistant nephrotic syndrome (SRNS) hyperlipidaemia may in the long term be associated with progressive renal insufficiency and increased risk of coronary heart disease. We have assessed the efficacy and tolerability of diet prior to and in combination with a hydroxymethylglutaryl CoA reductase inhibitor, simvastatin, in seven children with SRNS with a mean age of 8 years (range 1.8-16.3 years). Dietary advice to maintain adequate energy and protein intakes with reduced saturated fat and cholesterol intake had little impact on lipid levels pre treatment (mean reduction in cholesterol 1 mmol/l, triglyceride 1.1 mmol/l) but was maintained throughout the study duration. The mean cholesterol and triglyceride concentrations pre treatment were 12.1 +/- 2 (SEM) mmol/l and 8 +/- 2.1 (SEM) mmol/l, respectively. On a median simvastatin dose of 10 mg/day (range 5-40 mg) there was a 41% reduction in cholesterol to 6.6 +/- 0.77 (SEM) mmol/l and a 44% reduction in triglyceride to 3.9 +/- 1.38 (SEM) mmol/l at 6 months which was sustained at 12 months in five patients. The drug was well tolerated with no clinical side effects being noted. Over 6 months the mean plasma albumin concentrations increased from 18.2 +/- 1.26 (SEM) g/l to 23 +/- 2.51 (SEM) g/l, accounted for by three patients (1 complete remission, 1 partial remission, 1 end-stage renal failure). Plasma creatinine concentrations remained stable in five patients with two having progressive chronic renal failure. Growth parameters for both weight and height were maintained. Simvastatin has a beneficial effect on abnormal lipid levels in SRNS but the effectiveness of long-term therapy needs to be evaluated. PMID- 8703706 TI - Is there any reason to lower proteinuria in a child with nephrotic-range proteinuria who is not clinically edematous? PMID- 8703705 TI - Oral calcium loading test and response to diuretics in normal Taiwanese school children. AB - To investigate possible mechanisms of increased urinary calcium excretion and increased prevalence of urolithiasis in 16- to 20-year-old children, oral calcium loading and diuretic tests were performed in 120 normal children in three age groups (7-8, 12-13, and 17-18 years of age). Urinary calcium/creatinine ratios and 24-h urinary calcium excretion were significantly increased following the oral calcium loading test in 17- to 18-year-olds compared with the two younger age groups. Oral furosemide resulted in increased urinary calcium excretion in the 17- to 18-year age group, while hydrochlorothiazide was less effective in reducing urinary calcium excretion in this age group. These results suggest that increased intestinal calcium absorption and decreased renal tubular reabsorption of calcium in 17- to 18-year-olds may be contributing factors in the increased prevalence of nephrolithiasis in older Taiwanese children. PMID- 8703707 TI - Transforming growth factor alpha and epidermal growth factor expression in experimental murine polycystic kidney disease. AB - Cystic change in polycystic kidney disease (PKD) is associated with epithelial hyperplasia, altered fluid and electrolyte transport, and de-differentiation of renal tubular epithelium. The role of polypeptide growth factors as potential modulators of cystic change remains an area of controversy. In this study, the expression of epidermal growth factor (EGF) and transforming growth factor-alpha (TGF alpha) were assessed by immunohistochemistry and image analysis in glucocorticoid-induced PKD in the newborn mouse. Newborn C3H mice received either 200 mg/kg methylprednisolone acetate (MPA) or 0.9% saline as a control. EGF expression was not detected in significant quantities in either MPA-treated or control animals. TGF alpha, however, was expressed in immature control kidney in a largely basolateral distribution. Expression increased significantly in association with cystic change in MPA-treated animals and was localized to the apical cell surface, implying altered polarity of secretion. There is no evidence that EGF is a mitogen in this early developmental model of PKD. TGF alpha, however, may be an important mediator of cystic change in immature or de differentiated renal tubular epithelium. PMID- 8703709 TI - What are the present possibilities for the prenatal diagnosis of primary hyperoxaluria? PMID- 8703708 TI - Decrease in endothelin-1 renal receptors during the 1st month of life in the rat. AB - Endothelin-1 (Et1), like angiotensin II, is implicated in postnatal maturation and development. The present study was designed to identify Et1 receptors and subtype Et1 receptors present in rat kidney between 1 and 30 days of postnatal life. On day 1, high-affinity and high-density Et1 binding sites were identified in rat kidney. The dissociation constant and maximum binding for ET1 to membranes from whole kidney were 0.073 +/- 0.05 nM and 1,345.9 +/- 73 fmol/mg protein, respectively. On day 30, affinity and receptor density were markedly decreased. The dissociation constant and maximum binding were 0.147 +/- 0.021 nM (P < 0.01) and 633.2 +/- 56.4 fmol/mg protein (P < 0.001), respectively. Using BQ 123 (EtA selective antagonist) and sarafotoxin S6c (EtB-selective agonist), the two Et1 receptor subtypes EtA and EtB were identified in 1- and 30-day-old rat kidney. BQ 123 selectively recognized EtA receptors with high affinity (2.9 +/- 0.44 on day 1 and 4.0 +/- 0.5 nM on day 30) and sarafotoxin S6c bound with higher affinity EtB receptors (0.871 +/- 0.14 on day 1 and 0.717 +/- 0.12 nM on day 30). Between birth and day 30, the EtA binding capacity was decreased (304 +/- 27 vs. 752 +/- 202 fmol/mg protein, P < 0.05), whereas EtB binding was not affected (514 +/- 87 vs. 656 +/- 171 fmol/mg protein, NS). The decrease in the total number of Et1 receptors during the 1st month of life may be due to the concomitant decrease in the number of EtA receptors. Increased Et1 receptor density in early postnatal life suggests an influence of Et1 on immature kidney circulation and/or kidney growth. PMID- 8703710 TI - Histological long-term outcome of furosemide-induced nephrocalcinosis in the young rat. AB - The long-term prognosis of furosemide-associated nephrocalcinosis in the infant is still unclear. Although discontinuation of the diuretic often results in radiological resolution of the calcifications, functional abnormalities may persist. The natural history of the renal histopathology of these patients is yet unknown. In the present study we investigated the histological long-term outcome of furosemide-induced nephrocalcinosis in the young rat. Thirty-six weanling male Sprague-Dawley rats were divided into three groups: A controls, B furosemide given for 8 weeks, and C furosemide given for 2 weeks followed by 6 weeks of observation. Metabolic studies at the end of the experiment demonstrated a significant diuretic and natriuretic effect in group B. Kidney histology showed nephrocalcinosis scores (mean +/- SD) of 0.0 +/- 0.0 in A, 2.6 +/- 1.5 in B, and 0.8 +/- 0.6 in C, with B significantly higher than A and C, and C greater than A. Kidney calcium content in B (3,421.9 +/- 2,558.7 micrograms/g dry tissue) was significantly greater than in A (310.4 +/- 21.3) and C (1470.1 +/- 932.2). Another group of 6 rats receiving 2 weeks treatment of furosemide showed a nephrocalcinosis score of 2.2 +/- 1.5, not different from group B, and an additional group of 6 rats treated with furosemide for 2 weeks and observed for another 12 weeks showed a score of 1.3 +/- 0.4, not different from group C. We conclude that most of the renal calcifications induced by furosemide occur during the early days of treatment and that up to 12 weeks after discontinuation of the diuretic, the resolution of the calcifications is only partial. PMID- 8703711 TI - Total body water measurement in renal insufficiency. AB - Total body water was measured in 15 children with renal insufficiency (glomerular filtration rate < 25 ml/min per 1.73 m2) using deuterium oxide dilution. Total body water was also measured using bioelectrical impedance and skinfold anthropometry in the same 15 children. There was a linear correlation (r = 0.98, P < 0.01) between total body water measured by deuterium and height2/impedance. The 95% confidence limits for estimates of total body water were -1.5 to 0.9 for impedance and 0.65-3.16 l for skinfold anthropometry when compared with deuterium dilution. Bioelectrical impedance estimation of total body water is better than the current existing non-invasive method of skinfold anthropometry. PMID- 8703712 TI - Abnormal glomerular basement membrane in idiopathic multicentric osteolysis. AB - The primary cause of nephropathy in idiopathic multicentric osteolysis is as yet unknown. We report a young girl with idiopathic multicentric osteolysis and nephropathy. An abnormal glomerular basement membrane was the only abnormality found in a renal biopsy taken 2 years before the development of end-stage renal failure. We believe that this biopsy finding represents or is related to the unknown primary lesion causing nephropathy in idiopathic multicentric osteolysis. PMID- 8703713 TI - Cluster of cases of haemolytic uraemic syndrome due to unpasteurised cheese. AB - A cluster of four patients (1 girl, 3 boys) from a French village (2,000 inhabitants) had acute haemolytic uraemic syndrome (HUS) between March 1992 and May 1993. All had prodromes with fever and diarrhoea, then acute renal failure, anaemia, schistocytosis and thrombocytopenia. Peritoneal dialysis was carried out in three children (duration 3-12 days). The verotoxin VT2 gene was identified by polymerase chain reaction in the stools of two children. Some days prior to the diarrhoea, all children had eaten a cheese made with unpasteurised mixed cows' and goats' milk from the same farm. A case control study showed that the occurrence of HUS was linked to the consumption of this milk product (P = 0.006). The VT 2 gene was isolated from the cheese and from the stools of goats and cows from the farm, but not from the stools of farm employees. PMID- 8703714 TI - Superior sagittal sinus thrombosis in a child with nephrotic syndrome. AB - A 3-year-old male with steroid-responsive nephrotic syndrome developed a rare complication, sagittal sinus thrombosis during an episode of gastroenteritis, while on steroid therapy. Anticoagulation, as assessed by partial thromboplastin time, was difficult to maintain, despite administering high doses of heparin, infusions of fresh-frozen plasma to provide antithrombin III, and, subsequently, maximum doses of warfarin (0.3 mg/kg per day). Despite these problems the child made a complete neurological recovery. PMID- 8703715 TI - Retroperitoneal fibrosis and juvenile rheumatoid arthritis. AB - We describe a 13-year-old girl with juvenile rheumatoid arthritis who developed obstructive uropathy and renal failure. Retroperitoneal fibrosis (RPF) was confirmed by surgery. Although the renal failure and hydronephrosis resolved after surgery, the symptoms of vascular occlusion persisted. We consider that early diagnosis and treatment are essential. In cases of autoimmune disease, RPF should be considered when there is acquired obstructive uropathy accompanied by vascular occlusion syndrome. PMID- 8703716 TI - Protracted, gross hematuria in sickle cell trait: response to multiple doses of 1 desamino-8-D-arginine vasopressin. AB - Gross and microscopic hematuria are well-known complications in patients with sickle cell hemoglobinopathy. Most of these episodes of gross hematuria are self limiting, but rarely may be severe and persistent requiring definitive intervention. Before subjecting these patients to surgical management such as partial or total nephrectomy, several medical therapies of variable benefit have been suggested. We report a patient with sickle cell trait who experienced severe, intractable gross hematuria for 5 months and showed a dramatic response to multiple doses of 1-desamino-8-D-arginine vasopressin (DDAVP) infusion. The remarkable response observed in this patient suggests that treatment with DDAVP infusion may be considered in patients with unremitting gross hematuria associated with sickle cell trait. PMID- 8703717 TI - Respiratory syncytial virus infections in pediatric renal transplant recipients. AB - Immunocompromised patients are considered at increased risk from respiratory syncytial virus (RSV) infection. We examined the incidence and outcome of RSV infection in pediatric renal transplant (Tx) recipients on chronic immunosuppressive therapy. Of 173 recipients transplanted between November 1985 and April 1993, 5 (3%) developed RSV infection (age range 11-39 months). Initial immunosuppression included prednisone, azathioprine, cyclosporine, and polyclonal antibody therapy. Time from Tx to onset of RSV infection was 1 day to 7 months. Symptoms included rhinorrhea, cough, tachypnea, retractions, fever, wheezing, and abnormal chest X-ray. Treatment included bronchodilator therapy, bronchial drainage, ribavirin, and mist tent. Azathioprine was transiently withheld for leukopenia during treatment in 2 recipients. Three recipients developed biopsy proven acute rejection during (n = 2) or immediately following (n = 1) RSV infection; all responded to corticosteroid treatment. RSV infection is not commonly diagnosed in pediatric renal Tx recipients. The course of RSV infection in our patients did not differ from that reported in normal children. The possible association between RSV and acute rejection warrants further observation. When diagnosed early, RSV infection does not appear to be associated with increased mortality in pediatric renal Tx recipients. Larger numbers of recipients need to be studied to confirm these results. PMID- 8703718 TI - Loin pain haematuria syndrome. AB - Loin pain haematuria syndrome is a descriptive diagnosis of recurrent episodes of loin pain accompanied by haematuria, in which investigations do not reveal adequate pathology to account for the symptoms. The majority of patients present between 20 and 40 years, but onset may occur in older children. A significant number of patients show psychological and psychopathological features. Renal histology may show minor abnormalities, including mesangial proliferation, arteriolar and arterial hyalinosis and C3 in arterioles. Renal angiography is often normal but changes in intrarenal arterioles and cortical infarcts may be seen. Haematological abnormalities include decreased heparin-thrombin clotting time and elevated free plasma serotonin concentration. It is important that the assessment include a detailed psychiatric history, the patient's perception of pain, and the psychosocial environment. The pain may be very severe, leading to the requirement for addictive analgesics: management often becomes very difficult and frustrating to medical practitioners. Surgical intervention with capsulotomy, denervation and autotransplantation should only be considered as a last resort, as there is frequent recurrence of pain on the same or contralateral side. PMID- 8703719 TI - Chronic rejection and late renal allograft dysfunction. AB - Renal transplantation is currently standard therapy for end-stage kidney disease for children. Despite the considerable improvement in short-term results, the expected allograft half-life has remained the same. This is due to chronic rejection/late graft dysfunction which has proved resistant to therapeutic attempts. During the last few years the multifactorial pathogenesis of chronic renal allograft rejection has been clarified to some extent. Early injury by immunological and non-immunological mechanisms is followed by vascular remodelling due to repetitive cycles of cytokine release, upregulation of growth factors, and vascular smooth muscle cell proliferation. This leads to typical concentric arteriosclerosis and ischemia. Secondary kidney-specific mechanisms are initiated by the reduction in functioning renal mass and lead to gradual progression of chronic rejection. There is no single optimal therapy. Several attempts to influence the pathophysiological cascade have been promising. Attention should be focused on minimizing early immunological/non-immunological injury in order to attenuate future progression of chronic rejection. A significant prolongation of allograft half-life may be achieved during the next decade with the introduction of new therapeutic agents and comprehensive approach to treatment. This would be especially beneficial for pediatric recipients, reducing the need for retransplantation in adulthood. PMID- 8703721 TI - Proceedings of the American Society of Pediatric Nephrology Educational Symposium, San Diego, California, 7 May 1995. PMID- 8703722 TI - 26th annual conference of the European Working Group on Psychosocial Aspects of Children with Chronic Renal Failure. PMID- 8703720 TI - The current approach to the assessment of fetal renal function: fact or fiction? AB - The wide use of ultrasonography during pregnancy has led to the identification of a relatively large number of renal and urinary tract abnormalities, particularly hydronephrosis. Uncertainty, however, exists regarding the indications for surgical intervention. Prominent among the variables currently used to assess fetal renal function and predict the long-term outcome are measurements of urinary concentration of electrolytes and low molecular weight proteins. It has become, therefore, imperative to examine the evidence on which the selection of these variables is based, before they become entrenched in medical practice. The analysis reveals that single measurements of any of these variables is unlikely to be reliable. Sequential measurements, albeit more dependable, are impractical due to the narrow window of opportunity for useful surgical intervention. It is therefore necessary to search for new, innovative approaches aimed at detecting changes in the kidney or urine that reflect the biological response of the developing kidney to injury. Such markers may be represented by renal metabolites or peptides involved in the maintenance of renal function under conditions of stress. PMID- 8703723 TI - Clinical quiz. Genitourinary tuberculosis. PMID- 8703724 TI - A patient with Down's syndrome and anti-neutrophilic cytoplasmic antibody positive vasculitis. PMID- 8703725 TI - Treatment of childhood steroid-resistant nephrotic syndrome with pulse methylprednisolone and cyclophosphamide. PMID- 8703726 TI - A toddler with Down syndrome, hypercalcaemia, hypercalciuria, medullary nephrocalcinosis and renal failure. PMID- 8703727 TI - Progress towards poliomyelitis eradication. PMID- 8703728 TI - Expanded programme on immunization. PMID- 8703730 TI - On-going struggle. PMID- 8703729 TI - Influenza. PMID- 8703731 TI - Victim of his own success?. Interview by Graham Scott. PMID- 8703733 TI - Righting wrongs. PMID- 8703732 TI - Animal donors. PMID- 8703734 TI - Is sex safer? If you've heard one slogan, you've heard them all. PMID- 8703735 TI - The nursing challenge. PMID- 8703736 TI - Keeping up appearances. PMID- 8703737 TI - Bidding to build. PMID- 8703738 TI - The role of the nurse in medically oriented research. PMID- 8703739 TI - Family therapy: exploring the role of the CPN. AB - This article discusses the extent to which community psychiatric nurses (CPNs) practise family therapy. The author, after offering definitions of family therapy and discussing some of the models of the approach, suggests that although many CPNs will be conversant in the theory, only a minority feel confident to use in practice what is increasingly being seen as a specialisation in its own right. PMID- 8703740 TI - Total hip replacement: a study of customized prostheses. AB - The majority of widely used hip prostheses offer a limited range of offset sizes, including 35, 37, 40, 44 and 45mm. These sizes are provided to match the mean population offset which has been reported as 43.0mm (Noble et al 1988). The variability of proximal femoral geometry in terms of offset is consistent with the theory that the geometry of the femur is determined by a large number of genetic and environmental factors (Ericksen 1979). This paper suggests that the range in offset sizes offered by off-the-shelf hip protheses is not sufficient to match the extremes of anatomical offset variability of the Northern Ireland sample studied and that a custom stem that matches more accurately key femoral dimensions such as offset, may be more desirable. PMID- 8703741 TI - The sexual needs of elderly people: addressing the issue. AB - The subject of sexuality and elderly people has until recently, largely been ignored by nurses. This article stresses the importance of acknowledging that sexual activity and interest changes with ageing and that to deny that it exists is detrimental to the holistic care nurses should aim to provide. PMID- 8703742 TI - Analysing healthcare provision to ensure access for all. PMID- 8703743 TI - Woman prisoner shackled at her baby's funeral. PMID- 8703744 TI - Understanding and managing bereavement (continuing education credit). PMID- 8703745 TI - The learning profession. PMID- 8703746 TI - Leadership and you: planning for success. PMID- 8703747 TI - Leadership in practice: developing leadership in forensic mental health nursing. PMID- 8703748 TI - Meeting leadership and clinical practice development needs: a strategic approach. PMID- 8703749 TI - Leading today for tomorrow: the importance of nursing in the business of health care. PMID- 8703750 TI - Understanding leadership: challenges, changes and opportunities. PMID- 8703751 TI - Out of the trench: developing nurses' leaders. PMID- 8703752 TI - Take a balanced view. PMID- 8703753 TI - Going public. PMID- 8703754 TI - Duty-bound. PMID- 8703755 TI - All change. PMID- 8703756 TI - Help yourself to stop. PMID- 8703758 TI - Community child care: a study. PMID- 8703757 TI - Nursing the peace. PMID- 8703759 TI - Epidural analgesia: educating patients and nurses. AB - Epidural analgesia is used increasingly in operating theatres, delivery suites and intensive care units to manage post-operative pain. This article examines the importance of education in implementing a new procedure and explores the role of the anaesthetic nurse specialist. PMID- 8703760 TI - The introduction of mentorship to Project 2000 in Wales. AB - This study focused upon the introduction of mentors in the Common Foundation Programme (CFP) of Project 2000 (UKCC 1986) in Wales. It was commissioned by the Department of Health Research and Development Division on behalf of the Welsh Office Nursing Division. The study was policy oriented and its purpose was to inform future policy decision making through an analysis of the implementation of current policies for pre-registration education. The full title of our research project, 'The practitioner teacher: a study in the introduction of mentors in the pre-registration nurse education programme', implied that a clinically-based nurse practitioner with a designated teaching remit, fulfills a particular role (that of mentor) in the pre-registration nurse education programme. It was the nature, scope and impact of this mentor role during the initial implementation period of the CFP of Project 2000 in Wales to which this study addressed itself. PMID- 8703761 TI - Patient-nurse relationships: using reflective practice. AB - This article describes how a district nurse used a reflective journal to evaluate her practice when dealing with a tense patient-nurse relationship. Using a model of reflective practice and drawing on much of the theory on the subject, the author describes how the whole experience forced her to revise her perceptions of nursing. PMID- 8703762 TI - Bridging the research-practice gap: the role of the link nurse. AB - This article describes the gap between research and practice, and highlights an initiative at Rampton Hospital to bridge this gap through the introduction of ward-based link people. The authors outline the rationale behind the initiative and describe in detail how the strategy was put into practice. PMID- 8703763 TI - Venous leg ulcers (continuing education credit). PMID- 8703764 TI - Pushing the boundaries: nurse practitioner (continuing education credit). PMID- 8703765 TI - Going back to basics. PMID- 8703766 TI - Shame on us. PMID- 8703767 TI - Unequal health. PMID- 8703768 TI - The pension penalty for ME sufferers. PMID- 8703769 TI - The saga continues. PMID- 8703770 TI - Putting professional conduct in the dock. PMID- 8703771 TI - Listening and hearing. PMID- 8703772 TI - In the hot seat. Interview by Jean Gray. PMID- 8703773 TI - Use of compression stockings. PMID- 8703774 TI - General nurses' attitudes to patients who self-harm. AB - This article describes the prevalence of suicidal behaviour, both nationally and in relation to one inner city district general hospital. The authors discuss what is currently known about risk assessment for identifying those patients likely to repeat the act in the near future. They also examine the findings of a recent survey about nurses' perceptions and attitudes to patients who take deliberate drug overdoses. The survey found that nurses possessed some of the facts about risk factors relating to future parasuicidal intent and displayed generally professional attitudes to the treatment of these patients in general medical wards and A&E, but they appeared to show negative personal reactions after caring for patients who self-harm. PMID- 8703775 TI - End stage renal failure: the role of the nurse in patient education. AB - This article describes how a renal unit set up and implemented a structured patient education programme to help patients with chronic renal failure accept the nature of their disease, make choices about their treatment and adapt their daily life to cope with the situation. The author stresses the importance of information giving, to avoid what can be a frightening and depressing experience for patients. PMID- 8703776 TI - Diabetes mellitus at diagnosis (continuing education credit). PMID- 8703777 TI - 'Scope' brings liberation. PMID- 8703779 TI - Forward in time. PMID- 8703778 TI - Enrolled nurses worried over conversion plans. PMID- 8703780 TI - Not to blame. PMID- 8703781 TI - Gearing up for the election. PMID- 8703782 TI - Managing surgical wounds: a study report. PMID- 8703783 TI - Do birthplans empower women? A study of their views. AB - The birthplans has become a common practice in many maternity units to improve continuity of care and enable women to gain control over the experience of childbirth. In this study, semi-structured interviews were employed to explore the perception of empowerment from the women's point of view. Next week, the author highlights the midwives' experience of using birthplans. PMID- 8703784 TI - Nebuliser therapy: what nurses and patients need to know. AB - Optimal nebuliser therapy is achieved by selection of the appropriate nebuliser compressor system to target the delivery of a specific drug to a specific site in the lungs. Correct use of that delivery system by the patient converts optimal therapy into effective therapy. This article reviews what health professionals and patients need to know to fulfil that aim. PMID- 8703785 TI - Experiential learning and critical thinking in nursing. AB - This article describes a continuing ethnographic study of critical thinking, reflection and experiential learning in nursing. The author examines the related theory and explains the terms of reference which underpin the study. Date analysis so far emphasises the importance of recognising positive and negative feelings associated with experiences, when engaging in reflection. She also outlines the precursors to effective and non-effective reflection implied by the data. It is suggested that there is no correct model for critical thinking and the author highlights the need for further research into effective and non effective outcomes of reflection. PMID- 8703787 TI - Tissue viability. The use of compression hosiery in the care of leg ulcers. PMID- 8703786 TI - Tissue viability. Leg ulceration: a review of causes and treatment. PMID- 8703788 TI - Tissue viability. Evaluation of a leg ulcer clinic. PMID- 8703789 TI - Abdominal aortic aneurysm pathway: outcome analysis. AB - The current and future health care environment demands that health care providers place increased emphasis on the achievement of acceptable patient outcomes within an effective timeline and with more efficient use of resources. Clinical pathways provide a tool that defines the processes and activities that must occur to meet these goals. The purpose of this study is to describe a process for analysis of clinical and fiscal outcomes of a clinical pathway initiated at an academic medical center for elective abdominal aortic aneurysm repair. Patients were monitored throughout their preoperative and postoperative course to identify and trend variances, assess opportunities for improved resource use, and determine patient/family satisfaction. Data were collected by use of multiple electronic databases available within the university information systems network and analyzed to determine impact on patient charges, treatment course, and length of stay. Outliers were profiled as a means to identify prognostic indicators or establish a high preoperative risk. Results of a sample of 42 patients revealed a reduction in gross charges by 33% per case in comparison to the baseline data obtained before pathway implementation. This study will describe the rationale and process for instituting changes in resource use, such as diagnostic testing and blood use. Clinical outcomes and related nursing implications will also be discussed, including preoperative management, a streamlined same-day admission process, and factors associated with prolonged stay in the intensive care unit. Finally, strategies designed to enlist the support and participation of nurses, physicians, and other health team members will be discussed. PMID- 8703790 TI - Vascular trauma on the rise. AB - Trauma is the leading cause of death in people under the age of 45. Guns, drugs, and street violence contribute to this growing epidemic. As a result, there is an increased incidence of lower extremity vascular trauma. The most common cause of vascular injury is gunshot wounds. This article presents two case studies to illustrate the complex nursing management of this patient population. It is important for nurses to be aware of the physiologic and psychological treatment of these patients. A detailed nursing plan of care is also included. PMID- 8703791 TI - Heparin-induced thrombocytopenia. AB - Heparin is one of the most frequently prescribed medications in the United States with more than 1 trillion units being used each year. However, some patients who receive heparin may have development of a rare but severe complication called heparin-induced thrombocytopenia, which is associated with significant morbidity and mortality rates. This syndrome is a clinical event in which the use of heparin results in thrombocytopenia, venous or arterial thrombosis, and, less commonly, bleeding. The exact mechanism of heparin-induced thrombocytopenia is not clearly understood, but an immune-mediated response to heparin resulting in platelet consumption is the most widely accepted theory. The main treatment is discontinuation of heparin therapy; however, alternative treatments are discussed. The critical role nurses play in the early detection and prevention of the complications of heparin-induced thrombocytopenia is addressed. Assessment of all patients receiving any form of heparin therapy is emphasized. PMID- 8703792 TI - Undiagnosed hypercoagulable state: a case study. AB - Vascular nursing presents constant challenges. The natural history of vascular disease coupled with the complexity of the patient population provides an environment for constant learning. Providing care to these unique individuals demands a thorough knowledge of vascular anatomy and physiology, diagnostic interventions, treatment modalities, multidisciplinary resources, and nursing interventions. This case study explores the multifaceted realm of vascular nursing by examining the hospital course and multidisciplinary plan of care of a 22-year-old man whose hospital course began with a lower extremity deep venous thrombosis and progressed to pulmonary embolus, phlegmasia cerulea dolens, compartment syndrome, sepsis, arterial thrombosis, severe coagulopathy, priapism, laryngeal bleeding/laryngospasm, and subsequent notification of having received a unit of blood from a donor whose human immunodeficiency virus status was later determined to be positive. The intent of this article is not to define a specific means of practice but to share with colleagues the wealth of knowledge that was gained from this experience. PMID- 8703793 TI - Practicing nursing research. Help wanted? PMID- 8703794 TI - Ulnar artery aneurysm. PMID- 8703795 TI - Subclavian steal syndrome: a review. AB - The incidence of subclavian steal syndrome is poorly documented in recent literature; however, there is general agreement that it is a relatively uncommon diagnosis. The actual occurrence of a subclavian steal is more common than the associated syndrome. Symptoms of arm ischemia and vertebral-basilar insufficiency secondary to a subclavian steal can become quite disabling in some patients. Through astute observation and assessment, nurses may be the first to identify a patient with subclavian steal syndrome. Subclavian steal may be identified by nurses during routine history and physical examination. All patients initially seen with discrepant arm blood pressures or a diminished or weakened radial pulse unilaterally should have the diagnosis of subclavian steal considered during their evaluation. In addition, patients with vertebral-basilar symptoms or arm ischemia should have the diagnosis of subclavian steal ruled out. This article presents a review of the history, pathogenesis, current treatment modalities, and nursing care of patients with a subclavian steal. PMID- 8703796 TI - Challenges with Takayasu's arteritis: a case study. AB - Takayasu's arteritis (TA) is a systemic vasculitis that involves an autoimmune mediated transmural degeneration of the aorta and its major branches, leading to severe arterial stenoses or occlusions with subsequent cerebral, cardiopulmonary, mesenteric, renal, or limb ischemia. Patient history and physical examination, arteriography, magnetic resonance imaging, and specific laboratory tests facilitate the diagnosis of TA. Major treatments for the condition include the use of corticosteroids, cytotoxic agents, antiplatelet drugs, percutaneous transluminal angioplasty (PTA), and surgical revascularization. Major nursing interventions for TA include teaching the patient and family about the progression and treatments of TA and performing periodic patient assessments for manifestations of systemic vasculitis, alterations in tissue perfusion, and alterations in coping. PMID- 8703797 TI - Erythromelalgia--a case study. AB - Erythromelalgia is a rare syndrome characterized by pain, redness, and heat involving the lower and, less frequently, the upper extremities. Symptoms occur with local or environmental stimulation and may be mild for years or become disablingly severe. A finding of relief with ice-water immersion helps distinguish erythromelalgia from disorders such as causalgia and reflex sympathetic dystrophy. Erythromelalgia may be classified in three ways: (1) early onset, (2) adult-onset aspirin-sensitive, and (3) adult-onset non-aspirin sensitive. Treatment options, which include medication, sympathetic blocks, and surgery, are determined by classification. PMID- 8703798 TI - The Intermittent Claudication Research Study: vascular outcomes research using home health nurses. AB - This article describes an ongoing, multicenter outcomes research study on intermittent claudication being conducted at 16 Chicago-area vascular surgery offices and clinics. The study relies on gathering data through a series of home health follow-up visits by visiting nurses who use portable Doppler equipment. A total of 624 patients (506 men and 118 women) have been enrolled to date. Data on study patients' baseline demographic characteristics, leg symptoms, peripheral blood flow, comorbidities, and walking impairments are presented. Initial physical functioning and walking-distance scores from patient self-report questionnaires are also presented, along with attrition and lower-extremity revascularization rates. The contribution of nursing to this type of patient outcomes research is discussed. The changing climate in health care delivery provides nursing with a great opportunity to influence the U.S. health care system. In particular, the participation of nurses can make a major contribution to research on patient outcomes and medical effectiveness, and especially to new methods of comparing treatment effects on patients' functional status. Interpretation and measurement of patient self-reported health status and quality of life are particularly relevant to nursing, which is based on interventions related to compromised function. Improvement of patients' physical functioning is the heart of nursing practice, and the focus of much of what is done in vascular medicine and surgery. This article describes the contribution of nursing skills and knowledge to an ongoing, government-funded, multicenter research study on intermittent claudication. The ability of nurses to make a unique contribution to outcomes research is demonstrated. PMID- 8703799 TI - The place of blood stem cells in allogeneic transplantation. PMID- 8703800 TI - Graft-versus-leukaemia: understanding and using the alloimmune response to treat haematological malignancies. PMID- 8703802 TI - Effect of basic (FGF-2) and acidic (FGF-1) fibroblast growth factors on early haemopoietic cell development. AB - Basic fibroblast growth factor (FGF-2) and acidic fibroblast growth factor (FGF 1) are mitogens for a variety of cell types. Many reports suggest that haemopoietic cells are among these. Nevertheless, when we examined the effect of recombinant human FGF-1 or 2 on normal human marrow cell proliferation in vitro, only minimal stimulatory activity could be detected. In this regard, the addition of either growth factor to cultures of ancillary cell depleted marrow mononuclear cells (MNC), or to highly enriched CD34+ MNC, failed to enhance haemopoietic colony number and induced only a slight increase in colony size. Perturbation of FGF receptor (FGF-R) expression on CD34+ MNC with antisense (AS) oligodeoxynucleotides (ODN) was also without apparent effect on cell growth. Neither could we demonstrate any effect of FGF-1 or 2 on survival of early progenitor cells in serum-free culture. To explain these findings, we examined progenitor cells for expression of the FGF-R at the mRNA and protein level using RT-PCR and flow cytometry. Primitive CD34+/KIT+ MNC had no detectable FGF-R (FGF R1, 2, 3 or 4) mRNA or protein expression. In fact, direct immunofluorescence labelling of MNC for CD34 antigen and FGF-R1 demonstrated that expression of these markers was mutually exclusive in the populations examined. FGF-R1 expression was detected on subpopulations of MNC and on cells derived from day-6 CFU-GM and BFU-E colonies. Accordingly, FGF-R1 is either absent, or present at very low levels, on primitive haemopoietic cells. This fact, combined with our in vitro culture data, suggest that receptors are unlikely to play a significant role in the development of these early cells. Nevertheless, the development of mature cells may be influenced by the FGFs since the FGF-Rs are expressed on more mature cells. PMID- 8703801 TI - Protein kinase C isozymes in human megakaryoblastic leukemia cell line, MEG-01: possible involvement of the isozymes in the differentiation process of MEG-01 cells. AB - The expression of the different protein kinase C (PKC) isozymes in various states of differentiation of the human megakaryoblastic leukaemia cell line MEG-01 were analysed using thermocycle amplification of mRNA and immunoblotting. MEG-01 expressed mRNAs of PKC alpha, -beta I, -beta II, -delta, -epsilon, -eta, -theta and -zeta, but not PKC gamma. At the protein molecule level, MEG-01 was observed to express PKC alpha, -beta I, -beta II,- epsilon, -theta and -zeta, but lack gamma, -delta and -eta. When differentiation of MEG-01 was induced by 100 nm 12-O tetradecanoyl-phorbol-13-acetate (TPA), rapid translocation from cytosol to membrane fraction and down-regulation of PKC alpha, -epsilon and -theta was observed in 1-2h. On the other hand, PKC beta I and -beta II were observed to translocate not only to the membrane fraction but also to the cytoskeletal fraction in a different manner, and their down-regulation, especially beta II, was very slow. The myristoylated, alanine-rich C kinase substrate (MARCKS) in the membrane fraction of MEG-01 cells was observed to decrease gradually throughout the differentiation process. Additionally, time-course study of TPA treatment indicated that incubation of the cells for 30 min is sufficient for differentiation. These results strongly suggest that the activation of PKC alpha, -epsilon and -theta is involved in the initiation of differentiation, and that PKC beta I and -beta II have important roles in the maintenance of differentiation. Although PKC zeta was resistant to TPA treatment and its translocation was reduced, the amount of this isozyme in the cytosol fraction decreased throughout the differentiation process. PMID- 8703803 TI - A sensitive sandwich ELISA for measuring thrombopoietin in human serum: serum thrombopoietin levels in healthy volunteers and in patients with haemopoietic disorders. AB - A sensitive sandwich enzyme-linked immunosorbent assay (ELISA) has been established to estimate serum thrombopoietin (TPO) concentrations in healthy volunteers and patients with haemopoietic disorders. The ELISA uses a mouse monoclonal antibody (Ab) as the capture Ab and a biotinylated rabbit polyclonal Ab as the detector. The ELISA was reproducible, highly sensitive and specific for human TPO. The coefficients of intra-and inter assay variation were from 3.0% to 4.9% and from 5.9% to 6.1%, respectively. The quantitative limit of the ELISA was 0.09 fmol/ml in serum. The quantitative limit was lower than the normal level. The dose-response curves of serum samples from healthy volunteers and patients with haemopoietic disorders were parallel to the standard curves. The ELISA did not cross-react with a variety of blood components and cytokines to produce false positive results. The serum TPO concentrations from 29 normal males and 21 females were 0.79 +/- 0.35 and 0.70 +/- 0.26 fmol/ml, respectively. Serum TPO levels in patients with aplastic anaemia (AA), acute lymphocytic leukemia (ALL) and essential thrombocythaemia (ET) were measured using the ELISA. The serum TPO levels in the patients with ET (n = 6, 2.80 +/- 1.55 fmol/ml) were higher than the normal level. The patients with AA (n = 7, 18.53 +/- 12.37 fmol/ml) and ALL (n = 5, 10.36 +/- 5.57 fmol/ml) had significantly higher serum TPO levels than normal individuals. These results indicate that the ELISA specific to TPO should prove useful in measuring the TPO concentration in serum samples. PMID- 8703804 TI - Saporin, a ribosome-inactivating protein used to prepare immunotoxins, induces cell death via apoptosis. AB - The plant toxin saporin is a ribosome-inactivating protein which inhibits protein synthesis and growth of both normal and tumour cells. Its cytotoxic activity can be increased by coupling with antibodies recognizing cell surface antigens. In this work we performed experiments to test the hypothesis that saporin induces cell death via apoptosis. Exposure to saporin induced apoptosis in different cellular models, such as human peripheral blood B lymphocytes and neutrophils, in the Daudi B-cell line, and in the haemopoietic cell lines HL-60 and TF-1. This was indicated by: (a) the appearance of typical morphological features such as chromatin condensation, nuclear fragmentation and blebbing of plasma membranes: (b) DNA degradation into oligonucleosomal fragments: (c) the appearance of apoptotic cells on DNA flow cytometry as a cell population with reduced DNA content (A0 region). The fraction of cells showing features of apoptosis ranged from 19 +/- 5% for TF-1 cells to 35 +/- 8% for neutrophils. In experiments with normal peripheral blood B lymphocytes or with Daudi cells, we compared the activity of native saporin with that of an immunotoxin hybrid molecule obtained by binding the toxin to two bispecific antibodies recognizing both saporin and the B lymphocyte-specific antigen CD22 (Sap/BsAb complexes). Saporin bound to the antibodies was 2-3 logs more effective than native saporin in inducing apoptosis, with maximal inhibitions being observed at concentrations of 10(-6) M for native saporin and 10(-9)-10(-8) M for the hybrid molecules. These findings indicate that treatment with saporin results in apoptosis of target cells and suggest that this may be relevant to the therapeutic use of saporin-containing immunotoxins. In fact, if used in vivo as an immunotoxin, its cytotoxic activity could be devoid of more extensive and non-specific tissue damaging effects as would be the case if saporin induced necrosis of target cells. PMID- 8703805 TI - The role of sheep stroma in human haemopoiesis in the human/sheep chimaeras. AB - We have previously reported on the successful engraftment and long-term expression of human HSC (haemopoietic stem cells) in sheep following transplantation into pre-immune fetuses. The mechanism(s) underlying the support/maintenance of human haemopoiesis in sheep is not understood. In these studies we examined the ability of sheep stroma to support human haemopoiesis in vitro and compared this with support provided by human stroma. We found that, overall, human stroma was significantly more effective than sheep stroma in supporting human haemopoiesis in vitro. Nevertheless, stroma established from bone marrow of normal sheep was capable of lower-level support of the proliferation and multilineage differentiation of highly purified (CD34+, CD38-) preparations of human HSC in vitro. At lower concentrations, sheep stroma conditioned media (CM) induced human bone marrow mononuclear cells enriched for the CD34-positive fraction to enter the cell cycle at levels comparable to that of human stroma CM. However, unlike human CM, at higher concentrations, sheep CM inhibited human cells from initiating the cell cycle. These results provide a possible explanation for the long-term maintenance of low levels of human haemopoietic cell activity in human/sheep chimaeras. PMID- 8703806 TI - Hyper-interleukin (IL)-6-naemia in haemophagocytic lymphohistiocytosis. AB - Clinical features in patients with haemophagocytic lymphohistiocytosis (HLH) have been demonstrated to be characterized by hypercytokinaemia. Previously, we reported the impact of high serum levels of interferon (IFN)-gamma and soluble IL 2 receptor (sIL-2R) on patient outcome; however, it was not known if serum levels of interleukin (IL)-6 also could be a prognostic factor. In a study during the active phase of disease in 25 cases of HLH in children and young adults (median age 3 years, range 0.1-23 years), we noted 12 cases which showed serum IL-6 > 100 (normal < 4.0) pg/ml. Five of these cases showed hyper-IL-6-naemia alone without hyper-IFN-gamma-naemia (group A) whereas seven cases showed both hyper-IL-6- and IFN-gamma-naemia (group B). Patient outcome did not differ between the patients with IL-6 > 100 pg/ml and those with IL-6 < 100 pg/ml, suggesting that high serum concentrations of IL-6 alone do not necessarily indicate poor prognosis in patients with HLH. Among the cases with hyper-IL-6-naemia ( > 100 pg/ml), underlying disorders causing haemophagocytosis were found to be different between groups A and B. PMID- 8703807 TI - Haemophagocytic lymphohistiocytosis associated with constitutional inversion of chromosome 9. AB - Familial haemophagocytic lymphohistiocytosis (HLH) is considered an autosomal recessive disease, although the putative gene responsible for the disease has not yet been localized. Identification of the involved gene may elucidate the pathogenesis of the disease and is essential for prenatal testing in affected families. We present an infant with HLH and constitutional inversion 9 (p23q31) in cells from bone marrow, lymphocytes and fibroblasts. The parents had normal karyotypes. It may be speculated that one of the parents was a carrier of HLH and a de novo inversion occurred in chromosome 9 from the non-carrier parent. This would imply that the putative HLH-related gene is located at one of the two breakpoints on chromosome 9. PMID- 8703808 TI - Spectrum of diseases associated with increased proportions or absolute numbers of peripheral blood natural killer cells. AB - In a retrospective review of 1501 lymphoid flow cytometric studies of peripheral blood, we identified an increased proportion of natural killer cells in 125 cases (8%), 49 (3%) of which had a concomitant increase in absolute number of natural killer cells. Of the latter, the most frequent associated disorder was chronic natural killer cell lymphocytosis. Substantial quantitative increases in natural killer cells were also observed in some patients with lymphoma, leukaemia, immune thrombocytopenic purpura, or myelodysplastic syndrome. Our study provides incidence figures and clinical associations of an increased number of natural killer cells in the peripheral blood. PMID- 8703809 TI - Positive diepoxybutane test in only one of two brothers found to be compound heterozygotes for Fanconi's anaemia complementation group C mutations. AB - Fanconi's anaemia (FA) is an autosomal recessive disorder characterized by diverse congenital abnormalities, the development of progressive bone marrow failure, and an increased predisposition to malignancy, particularly acute leukaemia. The FA phenotype is so variable that diagnosis on the basis of clinical manifestations alone can be difficult. The modern diagnosis of FA no longer rests entirely on the constellation of clinical and haematological abnormalities first described by Fanconi, but depends on finding elevated chromosomal breakage after incubation of peripheral blood lymphocytes with the chemical clastogens diepoxybutane (DEB) or mitomycin-C (MMC). The cloning of the gene for FA complementation group C [FAC] provides an opportunity to test the validity of the "DEB test' which in recent times has become the main arbiter as to whether a patient is classified as FA or non-FA. We report on two brothers with similar clinical and haematological features who have both been identified as compound heterozygotes for the FAC mutations L554P and delta G322, but only one of the brothers has a positive DEB test. On the basis of the DEB test one would be classified as FA and the other as non-FA. The time has come to re evaluate the diagnostic criteria of "Fanconi's anaemia'. PMID- 8703810 TI - Tucaresol increases oxygen affinity and reduces haemolysis in subjects with sickle cell anaemia. AB - The primary pathophysiological event in sickling is the intracellular polymerization of deoxygenated haemoglobin S. Tucaresol (589C80;4[2-formyl-3 hydroxyphenoxymethyl] benzoic acid), a substituted benzaldehyde, was designed to interact with haemoglobin to increase oxygen affinity and has been shown to inhibit sickling in vitro. We administered tucaresol to sickle cell patients in the steady state to examine the anti-sickling effect in vivo. Oral doses of tucaresol or placebo were given to nine stable sickle cell patients (aged 17-39 years; tucaresol, six; placebo, three) for 10 d. The first two patients on tucaresol were scheduled to receive a loading dose of 800 mg or 1200 mg (depending on bodyweight) for the first 4 d, followed by maintenance doses of 200 or 300 mg for the next 6 d. Due to concerns over the sharp rise in haematocrit in one patient, subsequent cohorts received 300 mg tucaresol daily throughout the dosing period. The oxygen affinity of haemoglobin S was increased in all patients receiving tucaresol, with between 10% and 24% of the haemoglobin modified, dependent on dose. In all patients on tucaresol, haemolysis was reduced with rises in haemoglobin of 0.9- 3.7 g/dl (mean 2.2 g/dl), falls in lactate dehydrogenase of 16-52%, and a halving of the irreversibly sickled cell counts. These effects were apparent within a few days and persisted for 1-2 weeks following discontinuation of the drug. Three of the six patients on tucaresol developed fever and cervical lymphadenopathy, with onset between days 7 and 11 from start of drug. Further evaluation of the tolerability and efficacy of tucaresol in sickle cell patients is necessary. PMID- 8703811 TI - Contribution of haemolysis to jaundice in Sephardic Jewish glucose-6-phosphate dehydrogenase deficient neonates. AB - We determined the contribution of haemolysis to the development of hyperbilirubinaemia in glucose-6-phosphate dehydrogenase (G-6-PD) deficient neonates and G-6-PD normal controls. Blood carboxyhaemoglobin (COHb), sampled on the third day of life, was measured by gas chromatography, corrected for inhaled carbon monoxide (COHbC), and expressed as a percentage of total haemoglobin concentration (Hb). Serum bilirubin was tested as clinically necessary. 37 non jaundiced (peak serum total bilirubin (PSTB) < or = 255 mumol/l) and 20 jaundiced (PSTB > or = 257 mumol/l) G-6-PD-deficient neonates were compared to 31 non jaundiced and 24 jaundiced controls with comparable PSTB values, respectively. COHbC values for the entire G-6-PD deficient group were higher than in the controls (0.75 +/- 0.17% v 0.62 +/- 0.19%, P < 0.001). COHbC and PSTB values did not correlate in the G-6-PD-deficient group (r = 0.15, P > 0.05) but did in the controls (r = 0.58, P < 0.001). COHbC values were increased to a similar extent in the G-6-PD-deficient, non-jaundiced (0.72 +/- 0.16%), the G-6-PD-deficient, jaundiced (0.80 +/- 0.19%) and the control, jaundiced (0.75 +/- 0.18%) subgroups, compared to the control, non-jaundiced subgroup (0.53 +/- 0.13%) (P < 0.05). Although present in G-6-PD deficient neonates, increased haemolysis was not directly related to the PSTB. PMID- 8703812 TI - Ankyrin Napoli: a de novo deletional frameshift mutation in exon 16 of ankyrin gene (ANK1) associated with spherocytosis. AB - We report a case of apparently recessive hereditary spherocytosis in an Italian child. The proband exhibited a reduction of overall ankyrin in the red cell membrane. The parents were free of any haematological manifestations. The VNDR associated with the ankyrin gene (ANK1) were consistent with the following diplotypes: AC11/ AC14 (father), AC14/AC14 (mother) and AC11/AC14 (child). The cDNA of the patient disclosed the expression of the AC11 allele only. As a consequence, we put forward the hypothesis of a de novo inactivation affecting the ankyrin allele of maternal origin (AC14) and accounting for the disease. PCR amplification of exons, SSCP analysis and nucleotide sequencing disclosed a polymorphism: GAC --> AAC; Asp --> Asn in codon 328 of exon 10, and a one nucleotide deletion : CTG --> CG in codon 573 of the exon 16. This frameshift mutation placed in phase the TGA triplet that normally overlaps codons 636 and 637. Termination of translation near the middle of ankyrin mRNA coding sequence resulted, presumably, in its premature degadation. The present allele has been designated allele Napoli. PMID- 8703813 TI - A novel delivery system for continuous desferrioxamine infusion in transfusional iron overload. AB - Fifteen iron-overloaded thalassaemia major (TM) patients and two homozygous sickle cell patients (SCD) were treated continuously for 7d each week with the novel 48 h continuous subcutaneous (s.c.) desferrioxamine (DFX) delivery device (code C1083, Baxter) and 10 TM patients received the 24 h continuous intravenous (i.v.) DFX delivery device (code C1071). The 27 patients had previously received conventional s.c. DFX for 8-10h on 5 or more days each week. The serum non transferrin bound iron (NTBI) levels fell significantly in both groups within 12h of commencing the continuous infusion. In the s.c. group the mean level fell from 4.2 to 2.0 mumol/l (P = 0.001), whereas in the i.v. group the mean level fell from 3.6 to 0.1 mumol/l (P = 0.006) the initial levels being measured 12h after stopping conventional s.c. DFX. After 4 weeks there was a significant fall in serum ferritin in both groups (P = 0.009). The new DFX delivery device is effective at removing toxicfree iron from plasma and reducing body iron. Moreover, it is preferred by patients with much improved compliance compared to the conventional s.c. DFX pump. PMID- 8703814 TI - The frequency of the haemochromatosis-associated genotype D6S265-1:D6S105-8 in blood donors. AB - Homozygosity for HLA-A3 and the microsatellite markers D6S265 allele 1 and D6S105 allele 8 is associated with a high relative risk for genetic haemochromatosis indeed we and others have suggested that a haplotype including D6S265-1, HLA-A3 and D6S105-8 is specific for haemochromatosis. To determine the frequency of this haplotype and examine its specificity for haemochromatosis we have analysed data from 7820 blood donors from South Wales. The frequency of homozygosity for D6S265 1, HLA-A3 and D6S105-8 was 1 in 280. Calculations based on the prevalence of haemochromatosis suggest that about 50% of chromosomes carrying D6S265-1:HLA A3:D6S105-8 also carry the haemochromatosis gene. This information is of value for assessing the risk that the partner of a patient with haemochromatosis also carries the haemochromatosis gene. PMID- 8703815 TI - The dominant beta-thalassaemia in a Spanish family is due to a frameshift that introduces an extra CGG codon ( = arginine) at the 5' end of the second exon. AB - We have discovered a Spanish family with a dominant type of beta-thalassaemia. Carriers are characterized by mild anaemia, hypochromia, microcytosis, elevated Hb A2 and Hb F levels, reticulocytosis, and splenomegaly. The molecular basis of this condition is the introduction of a CGG triplet between codons 30 and 31 of the beta gene; this was determined by sequencing of amplified DNA and confirmed by dot-blot analysis. The abnormal mRNA (beta Th-mRNA) is stable and present in quantities similar to that of normal beta A-mRNA. cDNA fragments derived from beta Th- and beta A-mRNAs can be separated on a denaturing polyacrylamide gel electrophoresis because the beta Th fragment is three nucleotides (nts) longer than the beta A fragment. The beta Th-mRNA translates into a beta chain that is 147 amino acid residues long and carries an extra arginine residue between residues 30 and 31. This beta X chain has not been detected. It may be unstable and does not bind to the alpha chain. It probably is continuously digested by proteolytic enzymes in red cell precursors in the bone marrow. The abnormal chain probably binds haem that is excreted after proteolysis causing a darkening of the urine. PMID- 8703816 TI - A heteroplasmic point mutation of mitochondrial tRNALeu(CUN) in non-lymphoid haemopoietic cell lineages from a patient with acquired idiopathic sideroblastic anaemia. AB - Acquired idiopathic sideroblastic anaemia (AISA) has been proposed to be a disorder of mitochondrial DNA (mtDNA). The hallmark of mitochondrial iron overload may be attributable to a respiratory chain defeat leading to impaired reduction of ferric iron (Fe3+) to ferrous iron (Fe2+), which is essential to the last step of mitochondrial haem biosynthesis. In a 71-year-old patient we identified a point mutation in one of the two mitochondrial transfer-RNAs coding for leucine (tRNA(leu)(CUN)). The mutation involves a G --> A transition in the anticodon loop, immediately adjacent to the anticodon triplet (mtDNA position 12301). The mutated guanine is highly conserved in a wide range of species. The mutation is heteroplasmic, i.e. there is a mixture of normal and mutated mitochondrial genomes (ratio c. 50:50). Heteroplasmy of mtDNA is not found in normal individuals, but is a typical feature of mitochondrial cytopathies. The point mutation was present in the patient's bone marrow and whole blood samples, in purified platelets, and in the granulocyte/erythrocyte pellet after mononuclear cell separation by density gradient centrifugation. The mutation was not found in T- and B-lymphocytes isolated by immunomagnetic bead separation. It was also absent from buccal mucosa cells and cultured skin fibroblasts. This pattern of involvement suggests that the mutation occurred in a self-renewing myeloid stem cell of the CFU-GEMM type. PMID- 8703817 TI - Investigation of calmodulin and basic fibroblast growth factor (bFGF) in idiopathic myelofibrosis: evidence for a role of extracellular calmodulin in fibroblast proliferation. AB - The urinary concentration of calmodulin and basic fibroblast growth factor (bFGF) was determined in a total of 53 patients with various chronic myeloproliferative disorders (CMPD), including 22 patients with idiopathic myelofibrosis (IMF). Calmodulin excretion was significantly elevated in IMF (0.29 +/- 0.04 microgram/mmol creatinine) (P < 0.001), when compared to polycythaemia vera (PV) (0.14 +/- 0.02), essential thrombocythaemia (ET) (0.13 +/- 0.04), chronic myeloid leukaemia (CML) (0.16 +/- 0.02), unclassified myeloproliferative disorders (UMPD) (0.11 +/- 0.02) and age-matched controls (0.1 +/- 0.02) (P < 0.001). In contrast, bFGF was slightly elevated in all CMPD conditions when compared to age-matched controls. A neutralizing antibody to calmodulin was demonstrated to significantly influence the in vitro proliferation of normal human fibroblasts, an effect dependent on both cell density and the presence of fetal calf serum (FCS). Essentially, the antibody reduced FCS-induced proliferation of low-density fibroblasts but had little or no inhibitory effect on high-density fibroblasts in the absence of FCS. In addition, extracellular calmodulin was shown not to interact with known fibroblast mitogens, namely, IFG-1, EGF, bDGF and PDGF. We conclude that extracellular calmodulin should be considered, in addition to PDGF, TFG-beta and EGF, as a potential mitogen involved in the stromal reaction of idiopathic myelofibrosis. PMID- 8703818 TI - Mobilization of Philadelphia-negative peripheral blood mononuclear cells in chronic myeloid leukaemia using hydroxyurea and G-CSF (filgrastim). AB - A relatively simple and non-toxic out-patient-based regimen for the mobilization of Philadelphia-negative (Ph-ve) mononuclear cells in chronic myeloid leukaemia (CML) was evaluated in 10 patients, nine in stable chronic phase and one in accelerated phase. They received oral hydroxyurea at a mean dose of 3.5 g/m2 daily for 7 d, followed by 300 micrograms of G-CSF daily until the last day of harvesting. In the nine chronic-phase patients the mean number of days from the end of hydroxyurea to the commencement of harvesting was 14.5 (range 10-18). The patient in accelerated phase recovered and was harvested after 6 d. The mean number of aphereses performed was 3.4. Adequate numbers of stem cells were obtained in 9/10 patients judged by our usual criteria. Side-effects were mild in comparison to published intravenous schedules. No patients lost their hair. Five (50%) patients required admission with neutropenic fever which responded to antibiotics in all cases. Four (40%) patients developed a transient rash and four (40%) experienced mild oral mucostis. This level of toxicity enabled half of the patients to be treated entirely on an out-patient basis. The harvest products were analysed for cells belonging to the leukaemic clone by conventional cytogenetics, FISH and PCR. All were PCR positive. The mean Ph positivities by cytogenetics and FISH were comparable at 18.1% and 15% respectively. Half the patients had > 98% normal metaphases. We conclude that this approach is comparable in efficacy to published intravenous regimens and significantly less toxic. It can be safely used at diagnosis before interferon therapy commences. PMID- 8703819 TI - Evaluation of cytogenetic conversion to Ph- haemopoiesis in long-term bone marrow culture for patients with chronic myeloid leukaemia on conventional hydroxyurea therapy, on pulse high-dose hydroxyurea and on interferon-alpha. AB - Long-term bone marrow culture (LTBMC) has been used successfully in autologous transplantation in chronic myeloid leukaemia (CML). However, variation between patients in the recovery of Ph- cells in culture limits the application of this procedure to a minority. Treatment that effectively reduces in vivo tumour burden prior to initiation of LTBMC may improve the selection of Ph- cells in culture. To test this hypothesis we evaluated the frequency and degree of cytogenetic conversion to Ph- haemopoiesis in LTBMC from four independent groups of CML patients: Untreated (n = 19); conventional dosage of hydroxyurea (HU) (n = 10); pulse high-dose HU (P-HU) (n = 22) and interferon (IFN)-alpha (n = 12). In this study IFN-alpha therapy resulted in a significantly higher incidence of patients with detectable Ph- clonogenic cells in the marrow (P = 0.01) and with > or = 50% Ph- haemopoiesis in LTBMC as compared to newly diagnosed patients (P = 0.05). Also, sequential culture studies undertaken in 14 CML patients at diagnosis and following the start of pulse highdose HU therapy showed that in eight patients the average proportion of Ph- metaphases detected in LTBMC substantially increased from 1.7% (range 0-7) at diagnosis to levels of 71% (range 14-100) after treatment. Therefore we conclude that the use of IFN or pulse high-dose HU in early stage disease appears to create an opportunity to harvest the marrow for long-term culture (LTC) purging with reduced leukaaemic burden. PMID- 8703820 TI - Abnormal kinetics of colony formation by erythroid burst-forming units (BFU-E) in chronic myeloid leukaemia. AB - We have investigated the kinetics of colony formation by progenitor cells in chronic myeloid leukaemia (CML) using erythroid burst-forming units (BFU-E) as a model system. For this, we scored the numbers of subcolonies produced by individual BFU-E in cultures of normal marrow and blood cells and in cultures of CML blood cells. The formation of an erythroid burst consisting of a single subcolony was taken as evidence for immediate terminal differentiation; the formation of multiple subcolonies was taken as evidence for commitment to terminal differentiation only after several cell generations. Therefore the probability of differentiation can be obtained by scoring the numbers of subcolonies in individual erythroid bursts. We found that the probability of differentiation is decreased (P = 0.0004) and the number of subcolonies increased (P = 0.01) in CML BFU-E compared with normal BFU-E. The cellularity of the BFU-E was also increased in CML. Using the probabilities of differentiation and renewal obtained from the BFU-E cultures the results fitted the predictions of a stochastic branching model. These results indicate that (a) commitment to terminal erythroid differentiation occurs over several cell generations in populations of BFU-E, (b) the probability of commitment to terminal differentiation (PD) within a particular population of BFU-E, remains a constant independent of the number of cell generations involved, (c) PD is lower during burst formation by CML BFU-E than by normal BFU-E, and (d) commitment to terminal differentiation occurs over more cell generations in CML burst formation than in normal burst formation. Therefore a reduced probability of differentiation may be a primary defect and could explain the expansion of the erythroid progenitor cell compartment in CML. PMID- 8703821 TI - Platinum agents and secondary myeloid leukaemia: two cases treated only with platinum-based drugs. AB - With the increasing use of chemotherapy for many different primary malignancies, secondary or therapy-related acute myeloid leukaemias (AML) and myelodysplastic syndromes (MDS) are becoming more common. The risk of developing sAML has been estimated to be between 2% and 10%, depending upon the type, duration and dosage of previous therapy (Michels et al, 1985; Shulman, 1993; Robinson & Mertens, 1993; Ballen & Antin, 1993). It is therefore one of the most serious long-term complications of current cancer treatment and is likely to increase as longer survival rates for the primary tumour are achieved. An increasing range of drugs have been reported to cause sAML, including the alkylating agents, the epipodophyllotoxins and the anthracyclines, both as single agents and in combination (Pedersen-Bjergaard & Philip, 1991; Pedersen-Bjergaard & Rowley, 1994). We report two cases of secondary AML in which platinum compounds were the sole prior chemotherapy. PMID- 8703822 TI - Relation of blast cell survival and proliferation to chemotherapy resistance in AML. AB - We have investigated the in vitro blast cell survival (viability) and drug resistance to cytosine arabinoside (Ara-C), daunorubicin (Dau), mitoxantrone (Mitox) and aclarubicin (Acla) of fresh leukaemic blast cells from 80 patients with newly diagnosed acute myeloid leukaemia (AML) employing the semiautomated colourimetric MTT(3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) assay. In 15 cases we concurrently investigated the relation between in vitro blast cell survival (MTT assay) and blast cell proliferation (3H-thymidine incorporation) in the presence and absence of myeloid growth factors (GFs) G-CSF, GM-CSF and IL-3 (individually and in combination). A highly significant correlation was found between blast cell survival and blast cell proliferation (r = 0.87, P < 1 x 10(-4). Furthermore, in 40 evaluable adult patients who completed intravenous induction chemotherapy and were evaluable for response to chemotherapy we found a positive correlation between in vitro blast survival (MTT assay) and response to chemotherapy with high blast survival being associated with poor response to chemotherapy (P = 0.05). Moreover, in a multivariate analysis, high blast cell survival was significantly associated with high CD13 expression and monocytic phenotype (P = 0.0003 and P = 0.02, respectively). Furthermore, we found an inverse relationship between the baseline proliferation of the blasts and the magnitude of response to the GFs (P < 0.02), indicating that cells with low baseline proliferation were more readily stimulated by growth factors. Finally, we found a significant correlation between leukaemic cell survival and cellular drug resistance towards Dau (P = 0.001) and Mitox (P = 0.03), but not towards Ara-C (P = 0.68) and Acla (P = 0.13). We conclude that high in vitro leukaemic cell survival is associated with drug resistance in vivo and in vitro, and furthermore is correlated with high blast cell proliferation and some adverse prognostic factors previously identified in AML. PMID- 8703823 TI - Susceptibility of AML cells to in vitro apoptosis correlates with heat shock protein 70 (hsp 70) expression. AB - Acute myeloid leukaemia (AML) cells from some individuals rapidly undergo apoptosis during in vitro culture. We have analysed this mode of cell death in AML cells harvested from patients at initial presentation and during subsequent treatment/relapse. Using flow cytometric analysis of propidium iodide-stained cells and quantitation of the subdiploid apoptotic peak, we observed that leukaemic cells from patients with AML displayed a heterogenous susceptibility to apoptosis in terms of the rate of accumulation of apoptotic cells. After 48 h incubation in the absence of added serum or exogenous growth factors the percentage of apoptotic cells ranged from 3% to 99%. This susceptibility to apoptosis correlated significantly with intracellular expression of hsp70 (P = 0.009), but not hsp90, and was also associated with the presence of p53 and low levels of expression of bcl-2. PMID- 8703824 TI - In vitro resistance to cytosine arabinoside, not to daunorubicin, is associated with the risk of relapse in de novo acute myeloid leukaemia. AB - The efficacy of chemotherapy in acute myeloid leukaemia (AML) is limited by clinical drug resistance. We determined in vitro resistance to cytosine arabinoside (ARAC), daunorubicin (DNR), mitoxantrone (MITOX), m-amsacrine (AMSA) and etoposide (VP16) in 49 adults with de novo AML using the MTT assay. Results showed that nonresponders to chemotherapy were, in vitro, 2.9-fold more resistant to DNR, but not more resistant to ARA-C, compared to complete responders. However, complete responders who were in vitro resistant to ARA-C had a 4-fold higher risk of relapse (95% CI 1.3-12.5-fold) compared to complete responders in vitro sensitive to ARA-C. With a mean follow-up of 12 months the probability of continuous complete remission (CCR) for patients in vitro sensitive to ARA-C was 61% at 34 months (95% CI 28-82%), whereas all patients in vitro resistant to ARA C relapsed within 18 months from diagnosis. This difference appeared to be independent of other clinical features such as sex, age, white blood cell count, FAB classification, and CD34 expression. In vitro resistance to DNR was not related to the probability of CCR. We conclude that in vitro drug resistance assessed with the MTT assay appears to be associated with short- and long-term clinical outcome in AML. Confirmatory studies comprising a sufficient number of patients for multivariate analyses should prove whether in vitro resistance to ARA-C will appear to be an independent risk factor. PMID- 8703825 TI - Rearrangement of the BCL6 gene in B-cell lymphoid neoplasms: comparison with lymphomas associated with BCL2 rearrangement. AB - We report a series of B-cell neoplasms with regard to rearrangement of the BCL6 gene on chromosome band 3q27. Southern blot analysis using probes from the major translocation cluster (MTC) region of the BCL6 revealed rearrangement in 21/197 patients (10.7%) with B-cell neoplasms studied at presentation, and 11/25 patients (44%) first studied at relapse. In non-Hodgkin's lymphoma (NHL) studied at diagnosis, rearrangements of the BCL6 gene were not closely associated with a specific histopathologic subtype but distributed in subcategories in the Working Formulation. The incidence in follicular lymphoma was 12.1%, with significantly higher frequency in mixed and large cell subtypes, and that in diffuse aggressive lymphoma was 14.1%. Comigration analysis using probes from the immunoglobulin genes revealed association of the BCL6 gene with one of the three immunoglobulin loci in 9/25 cases analysed. A comparative study between NHL associated either with BCL2 or BCL6 rearrangement showed that advanced disease and bone marrow involvement were more frequent in BCL2(+) NHL. In contrast, extranodal involvement was more frequently observed in the BCL6(+) NHL. The survival curve of BCL6(+) NHL was characterized by a rapid decline followed by a plateau. Of the total of 32 BCL6(+) patients, six carried both BCL2 and BCL6 rearrangements; five of these six showed clinicopathological properties characteristic of follicular lymphoma, suggesting that the presence of the two genetic abnormalities does not necessarily have synergistic effects on malignant phenotypes. The high level of BCL6 expression in follicular lymphoma cell lines carrying a BCL2 rearrangement suggests that the deregulated BCL2 gene may have an effect on the development of genetic abnormalities of the BCL6 gene. The present study suggests that BCL6 gene rearrangement is primarily involved in large cell lymphoma irrespective of growth pattern of neoplastic cells, and that BCL6(+)BCL2(-) NHL could be curable with modern intensive chemotherapy. PMID- 8703826 TI - Differential expression of CD3 and CD7 in T-cell malignancies: a quantitative study by flow cytometry. AB - Most T-cell antigens are expressed on normal and neoplastic T lymphocytes and for this reason it is not easy to distinguish between the immunophenotype of normal and malignant T cells. We have addressed this problem by comparing the levels of expression of CD3 and CD7 on T lymphocytes from 18 healthy donors with those of 61 cases of T-cell leukaemia using quantitative flow cytometry with a method that converts fluorescence intensity into number of antigen molecules per cell. Normal T lymphocytes expressed 124 +/- 25 CD3 and 20 +/- 3 x 10(3) CD7 molecules per cell. The mean CD3 values were significantly lower in all types of T-cell leukaemia than in normal T cells (P < 0.05), with the exception of Sezary syndrome. The lowest CD3 values were found in T-lymphoblastic leukaemia (T-ALL), 30 +/- 21 x 10(3), and adult T-cell leukaemia/lymphoma (ATLL), 38 +/- 31 x 10(3), followed by T-prolymphocytic leukaemia (T-PLL), 92 +/- 47 x 10(3), and granular lymphocyte leukaemia (GLL), 95 +/- 21 x 10(3). In contrast, the number of CD7 molecules was significantly higher in T-All, 35 +/- 7 x 10(3) (P < 0.01), and T PLL, 29 +/- 12 x 10(3), than the normal controls (P < 0.01), whereas ATLL and GLL showed a low CD7 expression, 13 +/- 3 and 12 +/- 3 x 10(3), respectively. Our results show that the quantitative analysis of CD3 and CD7 and their combined evaluation may enable a distinction between normal and leukaemic T cells and could facilitate the monitoring of minimal residual disease. This study has also defined the T prolymphocyte as a cell of intermediate maturity between thymic derived and peripheral T lymphocytes. PMID- 8703827 TI - Expression of Fas antigen in acute myeloid leukaemia is associated with therapeutic response to chemotherapy. AB - Flow cytometric immunofluorescent analysis was used to assess Fas antigen (CD95) expression in blasts obtained from the bone marrow of 30 patients with acute myeloid leukaemia. The percentage of positive cells in each sample was highly variable. Fas antigen expression did not correlate with age, FAB subtype, white blood cell counts, or CD34 expression. Low expression of Fas was associated with a low complete remission rate after induction chemotherapy (62.5% in cases with < 20% positive cells v 92.9% in cases with > or = 20% positive cells, P < 0.01). The main cause for not achieving remission was resistant disease. Our results suggest that the quantitation of Fas expression can be predictive of treatment outcome in acute myeloid leukaemia. PMID- 8703828 TI - A phase I/II trial of Z-Dex (oral idarubicin and dexamethasone), an oral equivalent of VAD, as initial therapy at diagnosis or progression in multiple myeloma. AB - We designed an oral equivalent regime to mimic VAD and its hybrids, using idarubicin and dexamethasone (Z-Dex) given in four cycles to induce cytoreduction prior to dose intensification in multiple myeloma cases. 20 patients (de novo n = 15, replaced VAD n = 2, relapsed n = 2, and resistant n = 1), 13 males and seven females with a median age of 54 years (range 40-65 years) received Z-Dex therapy. The overall response rate was 70% (14/20), with one patient (5%) achieving complete remission (CR). The response rate for previously untreated patients was 80% (12/15), with a CR rate of 6.7% (1/15). Both patients who received Z-Dex in place of VAD continued to respond. Myelosuppression was seen in 14/20 patients (70%); 4/20 (20%) developing severe neutropenia with one death from neutropenic sepsis. Gastrointestinal toxicity and alopecia were infrequently reported. Satisfactory responses can be obtained using an oral regime equivalent to VAD with tolerable toxicity and morbidity. PMID- 8703829 TI - Pretransplant cytotoxic donor T-cell activity specific to patient HLA class I antigens correlating with mortality after unrelated BMT. AB - Unrelated bone marrow transplantation (BMT) is associated with increased post transplant complication rates, partly because more transplantation antigens are mismatched than in HLA-identical related BMT. We have shown previously that the cytotoxic T-lymphocyte precursor (CTLp) test performed before transplantation specifically detects HLA class I mismatches demonstrating its usefulness for the identification of new HLA class I alleles. In this study we analysed the clinical relevance of the CTLp test in 41 patients who underwent unrelated BMT between 1990 and 1994. All patient-donor pairs were HLA-A, -B, -DR compatible as defined by AB-serology and oligotyping for DR1-14. The host-reactive CTLp test was performed using previously frozen peripheral blood mononuclear cells (PBMC) as stimulators and PHA blasts as target cells. We found 10 CTLp-positive and 31 CTLp negative patient-donor pairs. Between the two groups there were no significant differences for age, diagnosis, sex, preconditioning and GvHD prophylaxis. The clinical results for the CTLp positive and the CTLp negative transplants were: severe acute GvHD III-IV 67% and 26% (P = 0.0315), transplant-related mortality 60% and 26% (P = 0.0085), and patient survival at 3.5 years 10% and 54% (P = 0.0006). Seven patient-donor pairs were mismatched for HLA-DR and/or -DQ subtypes. Only one of these seven class II mismatched pairs had a positive CTLp test. In the remaining nine CTLp positive pairs the CTL reactivity was directed against HLA-A, -B or -C antigens, revealing a statistically significant (P < 0.005) correlation between the CTLp frequency and HLA class I matching. In conclusion, the CTLp test helped to select optimally matched bone marrow donors and was particularly useful in association with high resolution oligotyping for DR- and DQ-subtypes for precise matching of both classes of HLA antigens. PMID- 8703830 TI - Duration of filgrastim mobilization and apheresis yield of CD34+ progenitor cells and lymphoid subsets in normal donors for allogeneic transplantation. AB - Seventy-seven normal donors underwent leukapheresis for peripheral blood progenitor cell collection beginning on day 4 (n = 45) or day 5 (n = 32) of filgrastim mobilization (12 micrograms/kg/d). The two groups were comparable for age, weight, blood volumes processed during leukapheresis and target CD34+ cell dose to be collected. The day 5 schedule allowed a more consistent achievement of the target cell dose with one apheresis (P = 0.005) and resulted in the initial collection of a significantly larger number of CD34+ cells (P = 0.009). There was no statistically significant difference in the leukapheresis yield of lymphoid subsets and natural killer cells. PMID- 8703831 TI - Acute-phase response and the hypercoagulable state in pulmonary tuberculosis. AB - In our experience, severe pulmonary tuberculosis (PTB) is often complicated by deep venous thrombosis (DVT). Because of the association between inflammation and haemostatic changes that can result in a hypercoagulable state, we have prospectively examined such predisposing factors in representative patients. Sequential analyses in a control group with active PTB showed anaemia, thrombocytosis, elevations in plasma fibrinogen, fibrin(ogen) degradation products (FDP), tissue plasminogen activator (t-PA) and inhibitor (PAI-1) with depressed antithrombin III levels. Age, sex and disease matched individuals with venographically proven DVT had higher FDP (15.8 +/- 14.3 v 3.2 +/- 1.7 micrograms/ml:P < 0.01), t-PA (19.4 +/- 14.9 v 11.3 +/- 0.8 ng/ml:P < 0.01), and functional PAI-1 activity (11.6 +/- 6.3 v 4.2 +/- 4.1:P < 0.01) with lower platelet counts (347 +/- 110 v 563 +/- 230 x 10(9)/1:P < 0.01). Fibrinogen levels in all patients rose during the first 2 weeks of therapy and, together with related disturbances, corrected within 12 weeks. In conclusion, elevated plasma fibrinogen with impaired fibrinolysis coupled with a decrease in antithrombin III and reactive thrombocytosis would appear to favour the development of DVT in PTB. PMID- 8703832 TI - Clearance of human factor XIa-inhibitor complexes in rats. AB - The serpins C1 esterase inhibitor (C1Inh), antithrombin (AT), alpha 1-antitrypsin (alpha 1AT) and alpha 2-antiplasmin (alpha 2AP) are known inhibitors of coagulation factor XIa (FXIa). Although initial studies suggested alpha 1AT to be the main inhibitor of FXIa, we recently demonstrated C1Inh to be a predominant inhibitor of FXIa in vitro in human plasma. The present study was performed to investigate the plasma elimination kinetics of preformed human FXIa-FXIa inhibitor complexes injected in rats. The amounts of complexes remaining in circulation were measured using enzyme-linked immunosorbent assays. The plasma half-life time of clearance (t1/2) was 98 min for FXIa-alpha 1AT complexes, whereas it was considerably shorter, i.e. 19, 18 and 15 min for FXIa-C1Inh, FXIa alpha 2AP and FXIa-AT complexes, respectively. Thus, due to this different plasma t1/2, preferentially FXIa-alpha 1AT complexes may be detected in clinical samples. Furthermore, measuring FXIa-FXIa inhibitor complexes in patient samples may not help to clarify the relative contribution of the individual serpins to inactivation of FXIa in vivo. PMID- 8703834 TI - The reduction of large von Willebrand factor multimers in plasma in essential thrombocythaemia is related to the platelet count. AB - We have investigated the relationship between platelet count and large VWF (von Willebrand factor) multimers in the plasma of 36 patients with essential thrombocythaemia (ET) and 26 patients with reactive thrombocytosis (RT). In both ET and RT patients an inverse relationship could be established between platelet count and large VWF multimers in plasma as well in relatively decreased ristocetin cofactor/von Willebrand factor antigen and collagen binding activity/von Willebrand factor antigen ratios. A normalization of the platelet count was accompanied by restoration of a normal plasma VWF multimeric distribution. Our data suggest that increasing numbers of platelets circulating in blood result in increased removal of large VWF multimers from plasma. PMID- 8703833 TI - Down-regulation of fibrinogen biosynthesis by IL-4, IL-10 and IL-13. AB - High levels of fibrinogen are recognized as an important vascular risk factor; however, it is not known if the increase of plasma fibrinogen is directly responsible for this risk, or is only a marker of vascular inflammation. To support this second hypothesis, Oncostatin M (OSM) is a potent stimulator of fibrinogen biosynthesis and induces smooth muscle cell proliferation. In the same way, we analysed whether interleukin-4 (IL-4), interleukin-10 (IL-10) or interleukin-13 (IL-13), which protect vessel walls from monocytes injuries leading to atherosclerosis, could influence fibrinogen biosynthesis. The two levels of regulation of fibrinogen biosynthesis were tested: firstly, the direct effect of these cytokines on fibrinogen production by the hepatoma cell line Hep G2, and secondly their effect on the secretion of hepatocyte stimulating factor (HSF) activity in the supernatant of lipopolysaccharide (LPS)-activated monocytes. IL-4 and IL-13 added to Hep G2 cells down-regulated both the increase of fibrinogen secretion induced by IL-6 and fibrinogen mRNA levels, this effect being more pronounced when Hep G2 were preincubated with the two cytokines before IL-6 addition. The effect of IL-10 was evidenced only on mRNA expression. IL-10 and IL-13 dose-dependently decrease HSF activity secreted by LPS-activated monocytes, whereas IL-4 had no effect. However, the three cytokines decreased HSF activity when monocytes were incubated with the cytokines before LPS activation. The effects of these cytokines on HSF activity are related to variations of IL-6 and OSM secretion. Our data strengthen the hypothesis that the fibrinogen level is a marker of vascular disease, since cytokines which have a protective vascular effect down-regulate fibrinogen production. PMID- 8703835 TI - Exon/intron structure of the human ALL-1 (MLL) gene involved in translocations to chromosomal region 11q23 and acute leukaemias. AB - The acute lymphoblastic leukaemia (ALL)-1 gene on human chromosome 11q23 is the site of many locally clustered chromosomal alterations associated with several types of acute leukaemias, including deletions, partial duplications and translocations. Structurally variant proteins derived from the altered gene presumably cause the malignant transformation of early haemopoietic progenitor cells. According to previously published reports, the gene consisted of at least 21 exons spread over approximately 100 kb. In this report a set of genomic fragments was isolated that represent a total of 35 exons (exons 3-37) encompassing > 95% of the protein-coding region (except exons 1 and 2) and the 3' non-translated region of the gene. The distances between these exons were determined and a detailed restriction map was produced. The majority of the exon/intron boundaries were sequenced and an intron-phase analysis was performed. The results form the basis for a greater understanding of the translocations and other structural alterations of the gene that conserve the open reading frame and thus produce presumably oncogenic variants of the ALL-1 protein. PMID- 8703836 TI - Inhibition of growth and induction of apoptosis by all-trans retinoic acid in lymphoid cell lines transfected with the PML/RAR alpha fusion gene. AB - The interaction of an exogenous PML/RAR alpha fusion gene associated with acute promyelocytic leukaemia, with all-trans retinoic acid (ATRA) was examined in two lymphoid cell lines. L1210 and MOLT-4 cells were transfected with PML/RAR alpha cDNA in the expression vector pGD and stable transformants (L1210PML/RAR alpha and MOLT-4PML/RAR alpha) were selected with G418. ATRA inhibited the growth of these stable transformants, as assessed by [3H]thymidine incorporation, in a dose dependent manner, but had no effect on the growth of control cells stably transformed with neomycin resistant gene alone. ATRA also induced apoptosis, as assessed by fragmentation of genomic DNA, in L121OPML/RAR alpha and MOLT-4PML/RAR alpha cells but not in control cells. The exogenous PML/RAR alpha fusion gene therefore probably mediates the effects of ATRA on cell growth and apoptosis in these cell lines. PMID- 8703837 TI - Pregnancy in women with essential thrombocythaemia. PMID- 8703838 TI - Myelodysplastic syndromes in children. PMID- 8703839 TI - Beyond the Conflict Tactics Scale: assessing gender differences in partner violence. AB - Previous studies of partner assault, particularly those using the Conflict Tactics Scales, have produced the controversial finding that women are as likely to assault their partners as are men. Such findings are clearly at odds with medical, legal, and social service agencies which find that women are far more often the victims of partner assault. Self-reported data from a national sample of young adults were used to determine the extent to which this apparent discrepancy could be reconciled. Results confirm previous findings of extensive violence by women, with little evidence of systematic over-or underreporting by either men or women. However, although both men and women engaged in frequent minor assaults, men were more likely than women to repeatedly beat their partner during the course of a year. In addition, women were far more likely than men to suffer physical injury and seek medical treatment as a consequence of incidents of male violence. Taken together, these findings somewhat reconcile the discrepancy regarding partner assault: women are more often than men the victims of severe partner assault and injury not necessarily because men strike more often, but because men strike harder. PMID- 8703840 TI - Childhood physical abuse, perceived social support, and socioemotional status in adult women. AB - This study investigated the extent to which an interactional model, relative to a main effect model, predicts the relationship between childhood physical abuse, perceived social support, and various aspects of socioemotional functioning in adult women. The results indicated that perceived social support during childhood was significantly related to subsequent levels of adult depression, trait anxiety, and child abuse potential in a manner consistent with a main effect model. Childhood history of physical abuse was related only to adult child abuse potential. Implications and study limitations are discussed. PMID- 8703841 TI - Interpersonal conflict which includes mistreatment in a university workplace. AB - In order to provide detailed documentation of faculty and staff experiences of perceived mistreatment, this article reports the findings of an institutional survey of mistreatment. From a random sample of 1,585 (11%) of one university's workforce, 810 (51%) responses were received. Of the total responses, 187 (23%) met criteria used to define cases of perceived workplace mistreatment. Equal proportions of women and men reported incidents of mistreatment. Mistreatment was perceived to come from coworkers as well as superiors. For the sample as a whole, job satisfaction was severely affected by experiences of mistreatment. The highest proportion of reported mistreatment was 38% for professional staff (at will employees), followed in order by 25% for classified staff (covered by civil service protection), and 11% for faculty. These findings indicate that mistreatment occurs at all levels in institutions of higher education. The negative consequences of mistreatment for individuals in the workplace, as well as for the institution as a whole, are substantial. PMID- 8703842 TI - A study of the prevalence of sexual coercion in adolescent heterosexual dating relationships in a Quebec sample. AB - Very few studies which have addressed the issue of sexual violence in dating relationships have provided prevalence rates for coercion that is perpetrated and sustained for both sexes during adolescence, while adequately accounting for the totality of unwanted sexual experiences. The present study addresses these issues by examining the rates of perpetrated and sustained sexual coercion among 644 French-speaking adolescents between 15 and 19 years of age with heterosexual dating experience from Quebec (Canada). Chi-square analyses reveal that girls are more often the victims of coercion, while boys more frequently perpetrate it. However, some boys are victims of sexual coercion. The most frequently occurring unwanted sexual experiences are kissing, and petting or fondling. Verbal coercion, consisting of continual arguments and pressure, is the strategy most often used by those who perpetrate unwanted sexual experiences, and is the strategy most frequently reported by those who suffer it. More extreme forms of sexual violence are also reported, but at a lower rate. Such high rates of sexual coercion during adolescence, as observed in the present study within a culture which is increasingly open to issues regarding sexuality, argue strongly for the implementation of prevention and intervention programs within this age group. PMID- 8703843 TI - Excuses, excuses: accounting for the effects of partner violence on marital satisfaction and stability. AB - For both theoretical and practical reasons, it is important to understand processes that lead to marital dissatisfaction and dissolution among women who are targets of relationship violence. Because attributional tendencies may often forecast marital behavior and because alcohol use is often seen as providing an excuse for deviant behavior, we examine two potential moderators of the associations between husband violence and wife marital outcomes: wife attributional style and husband problem drinking tendencies. A community sample of married couples (N = 66) completed a comprehensive battery of marital assessments. Results suggested that responsibility attributions moderated the association between husband violence and wives' marital dissatisfaction but exerted a direct effect on wives' disposition toward divorce. Husband problem drinking moderated the impact of husband violence only on wives' disposition toward divorce. As would be expected from an "excuse" model of the associations between violence and marital outcomes, violence had less of an impact on marital satisfaction and divorce ideation when wives attributed responsibility for negative spouse behavior as external to their husbands and when husbands were problem drinkers, respectively. PMID- 8703844 TI - Absolute and relative involvement in homicide offending: contemporary youth and the baby boom cohorts. AB - Recent concerns have been expressed that youths are an increasingly violent segment of U.S. society. This report explores such claims by presenting alternative dimensions with which trends in youth violence can be interpreted. Using Uniform Crime Reports and U.S. Bureau of the Census data for 1958-1993, rates of arrests for murder, taken to represent absolute levels of involvement in this form of violence, are analyzed for trends among 15-to 19-years-olds. Relative involvement, operationalized as the ratio of arrest rates for those aged 15-19 to those of the remainder of the population, is also analyzed for trends. A pronounced upward trend since the mid-1980s in both rates and ratios of arrests for murder is found for ages 15-19, resulting in this group now having the highest levels of absolute and relative involvement in murder arrests of any age category, a distinct departure from previous years. As a context for interpreting these levels, the involvement of current 15- to 19-year-olds is shown to exceed by a considerable margin the involvement of similarly aged cohorts of baby boomers, a youth group formerly the object of considerable public concern. Research is encouraged that addresses the multifaceted sources contributing to this dramatic societal shift in age-related patterns of arrests for murder and, by assumption, involvement in homicide offending. PMID- 8703845 TI - Recurrent gain of chromosome arm 7q in low-grade astrocytic tumors studied by comparative genomic hybridization. AB - Consistent tumor-specific chromosomal aberrations have not been described in low grade astrocytic tumors. The most frequent genetic alterations are mutations of the TP53 tumor suppressor gene and/or loss of heterozygosity (LOH) on 17p that occur in about 30% of the cases in adult patients but that are uncommon in childhood tumors. We used comparative genomic hybridization (CGH) to map DNA copy number alterations in 18 primary low-grade astrocytic tumors (ten adult patients and eight children). A gain of chromosome arm 7q was the most frequent event detected in five of ten astrocytomas (50%) from adult patients, followed by DNA amplification on chromosome arm 8q and gain on 12p (two cases). Loss of chromosomal regions on 1p, 4q, and the X chromosome was observed in two of ten cases [including one patient afflicted with Turner syndrome (45,X)]. In contrast, no consistent changes were observed in low-grade astrocytomas in children. A loss of the X chromosome was the sole aberration detected in two of eight cases using DNA extracted from the normal brain tissue. The findings suggest that a gain of 7q is an early event in the initiation of astrocytomas in adult patients. The discrepant findings in low-grade astrocytic tumors in adults compared to tumors in children support the the hypothesis that there might be different mechanisms responsible for tumor development. PMID- 8703846 TI - Analysis of the t(6;11)(q27;q23) in leukemia shows a consistent breakpoint in AF6 in three patients and in the ML-2 cell line. AB - The t(6;11)(q27;23) is one of the most common translocations observed in patients with acute myeloid leukemia (AML). The translocation breakpoint involves the MLL gene, which is the human homolog of the Drosophila trithorax gene, at 11q23 and the AF6 gene at 6q27. Reverse transcriptase-polymerase chain reaction (RT-PCR) using an MLL sense primer and an AF6 antisense primer detected the MLL/AF6 fusion cDNA from three leukemia patients with the t(6;11) [two AML and one T-acute lymphoblastic leukemia (ALL)] and one cell line. The fusion point in the AF6 cDNA from these cases is identical, regardless of the leukemia phenotype. The ML-2 cell line, which was established from a patient with AML that developed after complete remission of T-cell lymphoma, has retained an 11q23-24 deletion from the lymphoma stage and has acquired the t(6;11) with development of AML. The ML-2 cells have no normal MLL gene on Southern blot analysis, which indicates that an intact MLL gene is not necessary for survival of leukemic cells. PMID- 8703847 TI - Analysis of cyclin-dependent kinase inhibitor genes (CDKN2A, CDKN2B, and CDKN2C) in childhood rhabdomyosarcoma. AB - p16INK4A, p15INK4B, and p18 proteins are highly specific inhibitors of cyclin dependent serine/threonine kinase (CDK) activities required for GI-S transition in the eukaryotic cell division cycle. Mutations, mainly homozygous deletions, of the CDKN2A (p16INK4A/MTSI) gene have been recently found in tumor cell lines and in many primary tumors. We looked for homozygous deletions of CDKN2A, CDKN2B (p15INK4B), and CDKN2C (p18) in 12 primary rhabdomyosarcoma (RMS) specimens and in five cell lines established from this cancer type. By means of polymerase chain reaction (PCR) and PCR-single strand conformation polymorphism (PCR-SSCP), we analyzed the presence of biallelic gene deletion or point mutation causing gene function loss. All the examined tumor cell lines (100%) and three of 12 (25%) primary tumors showed homozygous deletion of CDKN2A. Furthermore, no aberrant bands in primary tumors were detected via SSCP, suggesting the absence of mutations in the coding region. In all cases the deleted area at 9p21 also involved the CDKN2B gene. Conversely, no homozygous deletion or point mutations were detected when CDKN2C was analyzed. Our results strongly indicate that the p16INK4A (and/or p15INK4B) protein plays a key role in the development and/or progression of childhood rhabdomyosarcoma and suggest that this CDK-inhibitor protein might control proliferation and/or differentiation of human muscle cells. Moreover, alteration of CDKN2C does not appear to be involved in the genesis of rhabdomyosarcoma. PMID- 8703848 TI - Allele loss on chromosome arm 6q and fine mapping of the region at 6q27 in epithelial ovarian cancer. AB - Genetic changes have been shown to be important in determining the multistep progression of cancer. Allele loss studies and karyotypic analysis of epithelial ovarian tumours have indicated the presence of putative tumour suppressor genes on chromosomes 6, 11, 13, 17, 18, 22, and X. We have focused on chromosome arm 6q to identify the minimal region that may contain a putative tumour suppressor gene. Nineteen polymorphic microsatellite markers from 6q and one centromeric marker (D6S294) have been used to detect allele loss in 68 ovarian tumours (six benign, six borderline, and 56 with malignant histology). Allele loss was evaluated by separation of fluorescence labelled polymerase chain reaction amplified products. Forty-six of fifty-six (82%) malignant tumours showed allele loss on 6q, whereas only four of 56 had lost all the markers tested. Forty-one of fifty-six (73%) malignant tumours showed allele loss at 6q26-27. The minimal region of allele loss was between markers D6S264 and D6S297 (3 cM), with maximal allele loss of 62% at D6S193 and 52% at D6S297. Three tumours showed loss of D6S193 only, while retaining flanking informative markers. Allele loss around 6q26-27 was observed in all histological types of epithelial ovarian cancer and did not correlate with any clinical factors. In addition, there was allele loss at ESR (56%) and D6S286 (47%), though a minimal region was not defined. Allele loss at 6q12-25 correlated significantly with endometrioid and mucinous ovarian malignant tumours (P = 0.01). The physical mapping of the region between D6S297 and D6S264 will allow the eventual identification of the putative tumour suppressor gene. PMID- 8703849 TI - Comparative genomic hybridization reveals a specific pattern of chromosomal gains and losses during the genesis of colorectal tumors. AB - Comparative genomic hybridization was used to screen the DNA extracted from histologically defined tissue sections from consecutive stages of colorectal carcinogenesis for chromosomal aberrations. No aberrations were detected in normal epithelium (n = 14). Gain of chromosome 7 occurred as a single event in low-grade adenomas (n = 14). In high-grade adenomas (n = 12), and overrepresentation of chromosomes 7 and 20 was present in 30% of the cases analyzed. The transition to colon carcinomas (n = 16) was characterized by the emergence of multiple chromosomal aberrations. Chromosomes 1, 13, and 20 and chromosome arms 7p and 8q were frequently gained, whereas chromosome 4 and chromosome arms 8p and 18q were recurrently underrepresented. The same tissue sections that were used for CGH were analyzed by means of DNA-ploidy measurements and immunohistochemical staining to quantify proliferative activity and p21/WAF-1 and TP53 expression. We observed that crude aneuploidy and increased proliferative activity are early events in colorectal carcinogenesis, followed by TP53 overexpression and the acquisition of recurrent chromosomal gains and losses during the progression from high-grade adenomas to invasive carcinomas. PMID- 8703850 TI - Variant translocation t(6;10)(p21;q22) in pulmonary chondroid hamartoma. AB - Banding cytogenetics and fluorescence in situ hybridization analysis of a pulmonary chondroid hamartoma (PHC) showed the presence of a t(6;10)(p21;q22). A cytogenetically identical translocation has previously been found in another case of PHC, suggesting that it could represent a variant form of the standard t(6;14)(p21;q24). PMID- 8703851 TI - Unilateral retinoblastoma--genetic implications. PMID- 8703852 TI - Activated protein C and retinal vein occlusion. PMID- 8703853 TI - Diagnostic assays in cytomegalovirus retinitis. PMID- 8703854 TI - Parents with unilateral retinoblastoma: their affected children. AB - BACKGROUND: A retrospective analysis of all patients with retinoblastoma on file at the Ophthalmic Oncology Center of the New York Hospital, Cornell University Medical Center was performed to examine the characteristics of retinoblastoma in children with a unilaterally affected parent. METHODS: Data were analysed for laterality of disease, age at initial diagnosis, number of tumour foci at diagnosis, ocular outcome, and patient survival. RESULTS: There were 54 patients, including nine pairs of siblings, diagnosed with retinoblastoma, each of whom had one unilaterally affected parent. The retinoblastoma was bilateral in 49 (91%) patients and unilateral in the remaining five (9%). The mean age at diagnosis was 10.5 months (range 1-36 months) and 35 patients (65%) were diagnosed within the first year of life. The mean number of initial tumour foci was 1.7 per eye; 1.6 foci per eye in bilaterally affected patients, and 3.7 foci per eye in unilaterally affected patients (p = 0.36). Retention of the eye was possible for 51 of 103 eyes affected with retinoblastoma. The mean follow up for patients was 9.0 years, with a range of 1 month to 30.5 years. There were a total of 12 deaths (22%): from retinoblastoma (two), sarcoma (five), cutaneous malignant melanoma (one), cancer type unknown (two), and cause unknown (two). CONCLUSION: Overall, the characteristics of retinoblastoma patients with a unilaterally affected parent, were similar when compared with all retinoblastoma patients with a positive family history. PMID- 8703855 TI - Activated protein C resistance in young adults with central retinal vein occlusion. AB - BACKGROUND: Central retinal vein occlusion is a disease that is most common in old people. While the disease in old people often is associated with atherosclerosis, hypertension, diabetes, or glaucoma, this is much less evident in young people. However, a new defect in the anticoagulant system has recently been discovered, activated protein C resistance. This hereditary defect may well be associated with central retinal vein occlusion, and so this factor was analysed in patients younger than 50 years with a history of central retinal vein occlusion. METHODS: Blood samples were obtained from 31 patients younger than 50 years with a history of central retinal vein occlusion, and analysed for activated protein C resistance with standard clinical laboratory methods. RESULTS: In this material 26% of all the patients and 36% of the patients younger than 45 years were resistant to activated protein C. The normal incidence of activated protein C resistance is 2-7%. CONCLUSION: Activated protein C resistance seems to be the most common known cause of central retinal vein occlusion in young people. PMID- 8703856 TI - Blood viscosity, coagulation, and activated protein C resistance in central retinal vein occlusion: a population controlled study. AB - BACKGROUND: The role of blood viscosity and haemostasis has been investigated in mixed groups of patients with branch and central retinal vein occlusion (CRVO) with conflicting results. This may have partly been due to the different aetiologies of these two types of vein occlusion. METHODS: In this study viscosity and coagulation (including activated protein C resistance) were examined in 87 patients with CRVO and compared with the results from an age matched, population based control group. RESULTS: Viscosity variables were higher in CRVO than in controls which suggested that reduced red cell deformability was associated with the occurrence of CRVO. A higher percentage of the patients with CRVO (12%) had activated protein C resistance than controls (5%). Patients who developed the complication of iris neovascularisation had relatively low antithrombin III, factor VII, and tissue plasminogen activator indicating both a tendency to thrombus formation and a reduction in fibrinolytic activity. CONCLUSION: Increased blood viscosity may contribute to the production of CRVO by inducing stasis of blood flow, with thrombus formation in at risk individuals who go on to develop iris neovascularisation. PMID- 8703857 TI - Hyperoxia improves contrast sensitivity in early diabetic retinopathy. AB - AIM: The cause of vascular and visual pathology in diabetic retinopathy remains unknown. If retinal hypoxia plays a role, then early in the course of diabetes 100% oxygen breathing should normalise both contrast sensitivity and retinal blood flow. METHODS: This hypothesis was tested in 12 diabetic patients with minimal retinopathy who, none the less, exhibited reduced contrast sensitivity (p = 0.003 versus 12 age and sex-matched controls) and prolonged retinal arteriovenous dye transit (p = 0.0001 versus controls). RESULTS: Isocapnic hyperoxia failed to alter contrast sensitivity in controls, while it significantly improved contrast sensitivity in patients (at 12 cpd; p = 0.042) to levels indistinguishable from normal. Individual improvement in contrast sensitivity correlated positively with the severity of the initial defect (r = +0.84, p = 0.0008). Hyperoxia also had haemodynamic effects; it slowed retinal arteriovenous passage of fluorescein dye in controls, but did not further slow this transit time in patients. CONCLUSIONS: These results demonstrate the reversibility of early contrast sensitivity deficits in diabetes mellitus, and support the hypothesis that factors linked to tissue hypoxia initiate both visual and vascular dysfunction in diabetic retinopathy. PMID- 8703858 TI - High dose intravitreal ganciclovir injection provides a prolonged therapeutic intraocular concentration. AB - BACKGROUND: Although intravitreal high dose ganciclovir has previously been found to provide excellent control of cytomegalovirus (CMV) retinitis, little was known about the vitreous concentrations of ganciclovir after a 2 mg intravitreal injection. METHODS: Eleven vitreous samples were taken from seven patients with CMV retinitis at 24 and 72 hours after a 2 mg intravitreal injection of ganciclovir and the concentration of ganciclovir was measured by high performance liquid chromatography. RESULTS: The mean concentration of ganciclovir at 24 hours was 143.8 mg/l (95% confidence interval 97.8-190) and at 72 hours was 23.4 mg/l (95% CI 0-49.7). The half life ranged from 11.9 to 26.3 (mean 18.8) hours in the four patients who had two samples taken. The mean half life calculated from the mean concentrations at 24 and 72 hours was 18.3 hours, so the calculated mean concentration at 7 days was 0.6 mg/l. CONCLUSIONS: This suggests that it takes about 7 days to eliminate the intravitreal ganciclovir, and that it is not likely to accumulate with weekly injections. The intravitreal concentrations achieved with high dose therapy remained above the ID50 for CMV (0.25-1.22 mg/l) for up to 7 days. PMID- 8703859 TI - Effect of isoproterenol, phenylephrine, and sodium nitroprusside on fundus pulsations in healthy volunteers. AB - AIMS/BACKGROUND: Recently a laser interferometric method for topical measurement of fundus pulsations has been developed. Fundus pulsations in the macular region are caused by the inflow and outflow of blood into the choroid. The purpose of this work was to study the influence of a peripheral vasoconstricting (the alpha 1 adrenoceptor agonist phenylephrine), a predominantly positive inotropic (the non-specific beta adrenoceptor agonist isoproterenol), and a non-specific vasodilating (sodium nitroprusside) model drug on ocular fundus pulsations to determine reproducibility and sensitivity of the method. METHODS: In a double masked randomised crossover study the drugs were administered in stepwise increasing doses to 10 male and nine female healthy volunteers. Systemic haemodynamic variables and fundus pulsations were measured at all infusion steps. RESULTS: Fundus pulsation increased during infusion of isoproterenol with statistical significance versus baseline at the lowest dose of 0.1 microgram/min. Neither peripheral vasoconstriction nor peripheral vasodilatation affected the ocular fundus pulsations. CONCLUSIONS: Measurements of fundus pulsations is a highly reproducible method in healthy subjects with low ametropy. Changes of local pulsatile ocular blood flow were detectable with our method following the infusion of isoproterenol. As systemic pharmacological vasodilatation or vasoconstriction did not change fundus pulsations, further experimental work has to be done to evaluate the sensitivity of the laser interferometric fundus pulsation measurement in various eye diseases. PMID- 8703860 TI - Effect of ablation profile on wound healing and visual performance 1 year after excimer laser photorefractive keratectomy. AB - BACKGROUND: Early photorefractive keratectomy ablations were of limited diameter and depth to maintain the integrity of the globe and to minimise postoperative haze. This study evaluated the effects of deeper, larger diameter wounds on refractive stability and corneal haze, and investigated the effects of ablation profile on wound healing and visual performance. METHODS: One hundred patients undergoing -3.00D and -6.00D corrections were randomised to receive 5 mm, 6 mm, or multizone treatments. The multizone treatment was 6 mm in diameter, but only the depth of the 5 mm treatment. Outcome was measured by Snellen visual acuity, residual refractive error, objective techniques for haze and halos, pupil diameter, subjective night vision, and requirement for retreatment. RESULTS: Overall, the results of 6 mm treatments were superior to those of 5 mm and multizone treatments: they had a smaller hyperopic shift (p < 0.01), a more predictable (p < 0.001) and stable refractive outcome, less haze (p < 0.05), smaller halos (p < 0.05), fewer subjective night vision problems, and fewer patients required retreatment. CONCLUSIONS: Analysis of these data and a literature review of corneal wound healing demonstrated that the improved outcome associated with the 6 mm beam did not relate to the depth of ablation. The factor with greatest apparent influence on the development of haze and regression was the slope of the wound surface over the entire area of the ablation. Tapering the wound edge provided no additional benefit, and contributed to night vision problems. It is, therefore, recommended that small diameter or multizone treatments should not be used in low and moderate myopia. PMID- 8703861 TI - Diagnostic assays in cytomegalovirus retinitis: detection of herpesvirus by simultaneous application of the polymerase chain reaction and local antibody analysis on ocular fluid. AB - AIM: To determine the value of the polymerase chain reaction (PCR) technique and the analysis of intraocularly produced antibodies by calculating a Goldmann Witmer quotient (GWq) as diagnostic assays in the confirmation of a clinically diagnosed cytomegalovirus (CMV) retinitis in a group of unselected AIDS patients. METHODS: Eleven samples of undiluted ocular fluid, obtained from nine AIDS patients with a clinically diagnosed CMV retinitis were analysed for the presence of genomic DNA from CMV, HSV-1, VZV, and EBV by PCR. Nine of these samples were analysed for the presence of locally produced IgG antibodies against these herpesviruses by calculating a GWq. Ten samples obtained from patients with various entities of clinical non-herpetic uveitis and 17 samples of aqueous humour obtained at cataract surgery were used as controls. RESULTS: In 10 out of 11 samples from AIDS patients (91%) the presence of CMV DNA was demonstrated. In four out of nine (44%) patients this was accompanied by CMV DNA in the blood indicating a CMV viraemia. In one sample, VZV DNA was detected and in another sample both CMV and VZV DNA were detected. No HSV-1 or EBV DNA could be demonstrated in these 11 samples. In contrast, simultaneous analysis of locally produced IgG antibodies against herpesviruses could not confirm the initial diagnosis of CMV retinitis. Ocular fluid samples obtained from 10 control uveitis patients were negative for DNA from CMV, VZV, and EBV by PCR. In one of 10 uveitis control samples HSV-1 DNA was detected; antibody analysis did not confirm this. In the uveitis control group, a significant GWq was calculated in one sample for HSV-1 and in another sample for VZV. The cataract control samples were all herpesvirus DNA negative by PCR. CONCLUSIONS: To establish the diagnosis of CMV retinitis in AIDS patients, ophthalmoscopic examination is a sensitive method. In confirming a diagnosis in indistinctive cases, application of a PCR assay detecting CMV DNA is a more sensitive method than analysis of locally produced antibodies by calculating a GWq. In clinical non-herpetic uveitis, secondary release of HSV-1 and VZV should be considered requiring additional therapeutic anticipation. PMID- 8703862 TI - Upregulated expression of vascular endothelial growth factor in proliferative diabetic retinopathy. AB - AIMS/BACKGROUND: Vascular endothelial growth factor (VEGF) is a hypoxia induced angiogenic factor. Recent studies have shown that high levels of VEGF accumulate in the vitreous of patients with proliferative diabetic retinopathy (PDR). The purpose of the present study was to identify the retinal cells that upregulate VEGF expression in human PDR patients representing progressive stages of retina deterioration. METHODS: Thirteen formalin fixed and paraffin embedded enucleated eyes with PDR were used (eyes were enucleated because of being blind and painful as a result of neovascular glaucoma). Thin retina sections were hybridised in situ with a VEGF specific probe, to identify cells producing VEGF mRNA. RESULTS: All eyes with PDR showed upregulated expression of VEGF mRNA, specifically in the cells of the neurosensory retina. VEGF expression was upregulated in all three nuclear layers--namely, the ganglion cell layer, the inner nuclear layer, and the outer nuclear layer. However, in each patient, VEGF producing cells were mostly distributed in a different layer, or even confined to a specific region in that layer. For example, expression by the outer nuclear layer was mostly detected in detached (presumably hypoxic) regions of the retina. CONCLUSIONS: Progression of PDR is distinguished by a sustained, upregulated expression of VEGF by the neurosensory retina. Cells in all retina layers can potentially contribute to augmented VEGF production. The restricted population of VEGF producing cells in each case is likely to represent cells residing in ischaemic regions of the retina. Thus, VEGF may function as a linking factor between retinal ischaemia and PDR associated neovascularisation. PMID- 8703863 TI - Three dimensional analysis of the retinal vasculature using immunofluorescent staining and confocal laser scanning microscopy. AB - AIM: To undertake a qualitative and quantitative analysis in three dimensions of the human retinal vasculature. METHOD: Fixed and excised whole retinas were permeabilised and subjected to immunofluorescent staining for blood vessel components followed by confocal laser scanning microscopy. Single projection and stereoimages were constructed using computer software. XZ sections through the retina were constructed and the vasculature analysed using appropriate software. RESULTS: Immunofluorescent staining with no discontinuities was present in vessels of all sizes, the confocal images of the capillary network being free of out of focus blur at all depths. Quantitative analysis of XZ sections confirmed the qualitative impression of sharp delineation of the deep retinal capillary plexus, an absence of laminar arrangement of capillaries within the inner retina, and a truncated cone of capillaries around the foveal avascular zone (FAZ) wherein the superficial capillaries approached the FAZ more closely than those in the deeper retina. CONCLUSION: Immunofluorescent staining of the retina and confocal laser scanning microscopy were shown to be useful in analysing accurate three dimensional reconstructions of the normal retinal vasculature without affecting overall tissue architecture. PMID- 8703864 TI - Ultrastructural features and prevalence of tubuloreticular structures in the ocular vasculature of patients with AIDS: a study of 23 cases. AB - BACKGROUND: Tubuloreticular structures (TRS) are subcellular inclusions that are most commonly found in endothelial cells and lymphocytes of patients with autoimmune or collagen vascular disorders. In AIDS, TRS have been described in various tissues throughout the body including the lung, kidney, liver, muscle, and skin. METHODS: Ocular tissues from 23 patients with AIDS were examined by electron microscopy. These included 17 postmortem eyes in addition to three chorioretinal and three conjunctival biopsy specimens. RESULTS: The overall prevalence of TRS in the ocular and conjunctival endothelial cells was found to be 83% (19/23). CONCLUSIONS: This is the first documented study of the prevalence of these structures in the ocular structures of patients with AIDS. Given the high frequency of their occurrence in AIDS, it is recommended that the presence of TRS in ocular or conjunctival tissues be an indication for obtaining an HIV antibody titre. Additionally, a rheumatological examination for HIV seronegative patients is suggested. PMID- 8703865 TI - Factors affecting bovine corneal endothelial cell density in vitro. AB - AIMS: To examine factors influencing the density and contact inhibition of bovine corneal endothelial cells cultured in vitro. METHODS: Cell counts were performed on bovine corneal endothelial cells cultured for various times in the presence of 10% fetal calf serum, with or without varying concentrations of growth factors, 5% dextran T-500, or 2% chondroitin sulphate, at 32 degrees C or 37 degrees C, and after treatment with beta galactosidase. RESULTS: Both basic fibroblast growth factor (FGFb) and retinal crude extract (RCE), but neither epidermal growth factor (EGF) nor acidic fibroblast growth factor (FGFa), increased endothelial cell density in vitro (p < 0.05). Continuous exposure to RCE resulted in a higher cell density than did a 24 hour pulse (p < 0.01), and higher cell densities were achieved at 37 degrees C than at 32 degrees C (p < 0.0001). In the absence of RCE, dextran T-500 increased cell density modestly (p < 0.05); in the presence of RCE, the addition of dextran T-500 had no effect on final cell density, whereas chondroitin sulphate significantly decreased final cell density (p < 0.01). In the absence of exogenous growth factors, beta galactosidase treatment resulted in a 50% increase in final cell density compared with controls (p < 0.0001). CONCLUSIONS: Bovine corneal endothelial cell growth can be augmented under conditions different from those used in corneal preservation systems. The final cell density in a confluent monolayer can be increased by treatment with beta galactosidase, suggesting that corneal endothelial cells may be contact inhibited through a beta galactosidase sensitive receptor system. PMID- 8703866 TI - Indocyanine green angiography. PMID- 8703867 TI - Benign fleck retina. PMID- 8703868 TI - Brown's syndrome as a complication of cardiopulmonary resuscitation. PMID- 8703869 TI - "In use" expiry date for eyedrops. PMID- 8703870 TI - Evidently, evidence based. PMID- 8703871 TI - The eye and the brain in non-accidental injury involving young children. PMID- 8703872 TI - The economic burden of global blindness: a price too high! PMID- 8703873 TI - Trends in the management of stage 3 retinopathy of prematurity. AB - AIMS/BACKGROUND: The clinical outcome of 66 consecutive infants with stage 3 retinopathy of prematurity (ROP) is reported. METHODS: Thirty four infants (64 eyes) were treated with cryotherapy and 32 infants (59 eyes) underwent laser photocoagulation. RESULTS: Infants with anterior-mid zone II ROP had a high rate of disease regression whether treated by cryotherapy or laser photocoagulation. Infants with posterior zone II or zone I ROP had a 40% success rate with cryotherapy, and 87.5% when treated with laser photocoagulation. CONCLUSION: Evolving experience and changing management policies in infants with posterior ROP has led to improved results. PMID- 8703874 TI - Ocular and cerebral trauma in non-accidental injury in infancy: underlying mechanisms and implications for paediatric practice. AB - AIMS: To determine the sites, mechanisms, and clinical significance of injuries to the eyes and brains of children with non-accidental injuries in relation to differing levels of trauma. METHODS: A forensic pathological study of injuries in the eyes and brains of 23 consecutive children dying of non-accidental injuries over a 4 year period (1988-92) under the jurisdiction of Yorkshire and Humberside coroners. RESULTS: Sixteen children died from cerebral injuries and seven died from non-cerebral causes. There were high incidences of retinal detachment (63%) and subhyaloid (75%), intraretinal (75%), and perineural (68%) haemorrhages in CNS deaths. Local subhyaloid haemorrhages and retinal detachment were more common at the periphery and optic disc than at the equator. There was a strong correlation between CNS and eye trauma scores in all 23 children (r = 0.7551, p < 0.0001). Ranking of injuries by severity suggests progressively more trauma required for (a) subdural haemorrhage, (b) subhyaloid, intraretinal, perineural haemorrhages, and (c) retinal detachment. At highest trauma levels choroidal and vitreous haemorrhages were associated with additional cerebral lacerations, intracerebral and subarachnoid haemorrhages. CONCLUSIONS: In non-accidental (and probably accidental) infantile head injury the earliest eye injuries (coinciding with subdural haemorrhage) could be missed if indirect ophthalmoscopy is not performed. Retinal detachment and multiple (particularly choroidal/vitreous) haemorrhages may indicate additional cerebral lacerations and/or intracerebral haemorrhage. Vitreous traction is the likely cause of intraocular pathology. PMID- 8703875 TI - Long term results of intraoperative mitomycin C in the treatment of recurrent pterygium. AB - AIM: The study was designed to evaluate the long term results of intraoperative mitomycin C in patients with one recurrence of pterygium. METHODS: In 45 white patients with one recurrence of pterygium the 'bare sclera technique' was performed and a sterile sponge soaked in a 0.2 mg/ml (0.02%) mitomycin C solution was placed intraoperatively on the sclera for 3 minutes. The control group underwent surgical excision only. Recurrences were analysed by the chi 2 test and the method of Kaplan-Meier (life table analysis); the difference between survival curves was tested by the log rank test. The chi 2 test with Yates's correction or Fisher's exact test were used to analyse the difference in complications and side effects between the two groups. RESULTS: After a mean postoperative follow up of 34.55 (SD 13.70) months, 6 recurrences (12.5%) were observed in the mitomycin C treated patients and 16 (35.6%) in the control patients (p = 0.027). The 24 and 48 month life table success rates were 89% and 83% in the mitomycin C treated group and 66% and 63% in the control group, respectively (p = 0.022). No severe side effects appeared during follow up. Superficial punctate keratitis appeared in the early postoperative period in only seven mitomycin C treated eyes (15.5%) (p = 0.018). CONCLUSION: This study confirms the efficacy of intraoperative mitomycin C in improving the success rate after recurrent pterygium surgical excision. PMID- 8703876 TI - Progression of disease in ocular cicatricial pemphigoid. AB - BACKGROUND: Ocular cicatricial pemphigoid (OCP) is a sight threatening autoimmune disease that can lead to severe conjunctival cicatrisation and keratopathy. It has a variable course and little is known about the factors that determine disease progression. This study analysed the factors that have prognostic significance regarding disease outcome, progression, and keratopathy. METHODS: Sixty six patients with OCP were monitored prospectively at Moorfields Eye Hospital. The influence of ocular features, the systemic disease, and the management were analysed to identify factors that influence the outcomes and disease progression. RESULTS: The mean age at presentation was 67 years; 56% were men. The binocular visual acuities were 6/24 or worse in 25%. Extensive cicatrisation at presentation was common but correlated only weakly with the visual prognosis. Systemic manifestations included lesions of the mouth in 44%, pharynx in 30%, oesophagus in 27%, nose/sinus in 18%, and skin in 17%. There was no association between the ocular and systemic manifestations. Persistent corneal epithelial defects and limbitis occurred in 18% and 32%, respectively, and both were associated with a worse visual prognosis. Systemic immunosuppression was ultimately prescribed in 74%, mainly in patients with advanced stages of conjunctival cicatrisation. Of patients with more than 24 months follow up, progression of cicatrisation occurred in 35% of eyes (16/46) all but one of which were associated with episodes of conjunctival inflammation. CONCLUSIONS: Persistent epithelial defects, limbal inflammation, and ongoing conjunctival inflammation are important factors that lead to keratopathy and visual handicap. These require aggressive management, often with systemic immunosuppressive treatment. Close follow up is required in cases with extensive cicatrisation. PMID- 8703877 TI - Objective detection of hemifield and quadrantic field defects by visual evoked cortical potentials. AB - AIMS/BACKGROUND: An objective method for detecting hemifield and quadrantic visual field defects has been developed using steady state visual evoked cortical potentials (VECPs), an adaptive noise canceller (ANC), and Hotelling's t2 statistic. The purpose of this study was to determine the sensitivity and specificity of the technique. METHODS: Nine subjects (mean age 44 years) were investigated with field loss due to a variety of causes including both anterior and posterior visual pathway lesions. Dynamic perimetry was performed by means of a Goldmann or Tubingen perimeter. VECP recordings were made from each visual field quadrant (23 degrees X 23 degrees) by means of a steady state reversing checkerboard (7.7 rev/s). The central 5 degrees of the visual field and the vertical and horizontal meridians were masked during these measurements. Recordings were made from three electrode sites, positioned over the visual cortex, relative to a mid frontal electrode. Each recording lasted 2 minutes, during which time fixation was monitored. The data from each recording were divided into 4 second segments, and the amplitude and phase of the VECP signal measured using the ANC. Hotelling's t2 statistic was applied to determine the probability of signal detection. Receiver operating characteristic curves were used to find the optimum signal detection threshold for identification of the visual field defects. RESULTS: The results of the study confirmed patterns of subjective visual field loss. The technique had a sensitivity and a specificity of 81% and 85%, respectively, for detecting 'non-seeing' areas in the inferior visual field, and 82% and 89%, respectively, for detecting 'non-seeing' areas in the superior visual field. CONCLUSION: These results demonstrate that the technique is of potential clinical value to ophthalmologists and neurologists when subjective perimetry is not possible. PMID- 8703879 TI - Automated perimetry in patients with choroidal metastases. AB - AIMS/BACKGROUND: Recent work using computerised perimetry has documented predictable visual field defects in patients with choroidal melanoma. Despite the higher frequency of occurrence of choroidal metastases, relatively little is known about their exact effects on visual performance, specifically with respect to visual field. This study is a pilot retrospective analysis of the results of automated perimetry testing in patients with choroidal metastatic disease. METHODS: The Humphrey field analyser was used to perform visual field tests on 15 eyes in 11 patients diagnosed with choroidal metastases. All 11 primary tumours were carcinomas originating in the breast (seven), lung (one), kidney (one), stomach (one), and prostate (one). All patients had either central 30 and/or peripheral 30-60 threshold strategies. RESULTS: Each of the 15 eyes tested had a demonstrable visual field defect. Eight eyes (53%) had absolute scotomas and seven (43%) had relative scotomas; the defects did not consistently correspond in size or location to the tumour as depicted by ophthalmoscopy or ultrasonography. This is in contradistinction to automated perimetry results in patients with choroidal melanoma. Three of the four eyes (75%) which were retested after treatment of the metastases showed improvement of the visual field. Seven of nine eyes (77%) that initially had reduced visual acuity had improvement after treatment. This clinical improvement corresponded well to tumour shrinkage and resolution of subretinal fluid. CONCLUSION: These findings may be useful in the decision to initiate treatment for symptomatic choroidal metastatic disease and in explaining to patients and their referring physicians what improvement could be expected from such treatment. A prospective evaluation of patients with choroidal metastases seems appropriate based on this pilot experience and may elucidate if the differences noted between visual field performance of these patients and those with choroidal melanoma may be useful in some diagnostically difficult cases. PMID- 8703878 TI - Visual field test simulation and error in threshold estimation. AB - AIM: To establish, via computer simulation, the effects of patient response variability and staircase starting level upon the accuracy and repeatability of static full threshold visual field tests. METHOD: Patient response variability, defined by the standard deviation of the frequency of seeing versus stimulus intensity curve, is varied from 0.5 to 20 dB (in steps of 0.5 dB) with staircase starting levels ranging from 30 dB below to 30 dB above the patient's threshold (in steps of 10 dB). Fifty two threshold estimates are derived for each condition and the error of each estimate calculated (difference between the true threshold and the threshold estimate derived from the staircase procedure). The mean and standard deviation of the errors are then determined for each condition. The results from a simulated quadrantic defect (response variability set to typical values for a patient with glaucoma) are presented using two different algorithms. The first corresponds with that normally used when performing a full threshold examination while the second uses results from an earlier simulated full threshold examination for the staircase starting values. RESULTS: The mean error in threshold estimates was found to be biased towards the staircase starting level. The extent of the bias was dependent upon patient response variability. The standard deviation of the error increased both with response variability and staircase starting level. With the routinely used full threshold strategy the quadrantic defect was found to have a large mean error in estimated threshold values and an increase in the standard deviation of the error along the edge of the defect. When results from an earlier full threshold test are used as staircase starting values this error and increased standard deviation largely disappeared. CONCLUSION: The staircase procedure widely used in threshold perimetry increased the error and the variability of threshold estimates along the edges of defects. Using earlier data, when available, overcomes this problem and reduces examination time. PMID- 8703880 TI - Postoperative abnormalities of the choriocapillaris in exudative age-related macular degeneration. AB - AIMS: To study the incidence and possible cause of abnormalities of the subfoveal choriocapillaris after surgical excision of subfoveal choroidal neovascularisation in age-related macular degeneration (ARMD). METHODS: The postoperative fluorescein angiograms and colour photographs of 29 eyes of 29 patients were reviewed after surgical excision of subfoveal choroidal neovascularisation in exudative ARMD. Preoperative and postoperative fluorescein angiograms were examined for perfusion of the subfoveal choriocapillaris. The excised subfoveal choroidal neovascular membranes from eight eyes that demonstrated postoperative abnormalities of the choriocapillaris were embedded in paraffin, serially sectioned and examined for the presence of the choriocapillaris. RESULTS: Postoperative fluorescein angiograms revealed abnormal perfusion of the subfoveal choriocapillaris in 26 of the 29 eyes (90%) and in all eight eyes that had histopathological examination of the surgical specimens. Examination of serial sections demonstrated that none of the excised neovascular membranes contained choriocapillaris. CONCLUSIONS: Abnormal perfusion of the subfoveal choriocapillaris was frequently present following removal of the subfoveal neovascular membrane in ARMD. The histopathological study demonstrated that abnormalities of the choriocapillaris were not due to removal of the choriocapillaris at the time of surgery. PMID- 8703881 TI - Do you have a Roman connection? PMID- 8703882 TI - Comparative effects of argon green and krypton red laser photocoagulation for patients with diabetic exudative maculopathy. AB - AIMS/BACKGROUND: Focal treatment of diabetic macular oedema is usually done using a haemoglobin absorbing wave-length, such as argon green laser. This study aimed to compare photocoagulation with argon green (514 nm) and krypton red (647 nm), which is poorly absorbed by haemoglobin, in the focal treatment of patients with diabetic exudative maculopathy. METHODS: A total of 151 eyes of 78 outpatients were assigned randomly to receive either argon green (n = 79) or krypton red (n = 72) laser treatment. Pretreatment and post-treatment ocular examinations included visual acuity, fundus biomicroscopic examination, and fluorescein angiography. A total of 141 eyes of 73 patients were available for evaluation after 1 year follow up. RESULTS: No statistically significant difference was found between the two treatment groups with respect to visual acuity results, resorption of hard exudates, and resolution of focal retinal oedema. CONCLUSION: Krypton red laser was effective in the treatment of diabetic exudative maculopathy. Our data suggest that differential absorption of the various wavelengths by haemoglobin within microaneurysms may not be an important factor for ultimate success of focal treatment. PMID- 8703883 TI - Results of surgery for epiretinal membranes and their recurrences. AB - AIMS/BACKGROUND: This study was performed to identify factors predictive for recurrence of idiopathic and secondary epiretinal membranes after vitrectomy. Long term visual outcome was determined and compared with eyes without recurrence of epiretinal tissue. METHODS: Out of 350 consecutive cases vitrectomised for removal of epiretinal macular membranes, 42 patients (42 eyes) with recurrence of epiretinal membranes were re-examined and their records reviewed. Recurrence of epiretinal membranes was determined by new appearance of epiretinal tissue or abnormal sheen of the central retina. The preoperative findings, best postoperative and final visual acuity of these patients were compared with a comparable cohort of eyes without recurrence after removal of macular pucker (131 eyes). The eligibility criterion was a follow up of at least 6 months. RESULTS: The best postoperative (0.46 (SD 0.24)) and the final visual acuity (0.39 (0.26)) of eyes with recurrent membranes (42 eyes) was statistically better than preoperative vision (0.27 (0.22)) (p = 0.0003 and 0.0089). Because of reduced or distorted vision eight of the 42 eyes underwent a revitrectomy during the follow up (mean 23 months). Best postoperative visual acuity (0.50 (0.21)) and final visual acuity (0.47 (0.25)) of eyes which received a second vitrectomy (n = 8) were not statistically different from the 32 eyes with recurrence of epiretinal tissue but without revitrectomy (p = 0.253 and 0.343). In addition, eyes with recurrence showed no different visual outcome compared with eyes without recurrence of epiretinal membranes after vitrectomy (p = 0.84). Recurrence of epiretinal membranes was correlated with neither the type of membrane (thick or thin) nor incomplete membrane removal during pars plana vitrectomy. CONCLUSION: This study demonstrates the overall favourable visual prognosis of patients with vitrectomy for macular pucker, even with recurrence of the premacular membrane. PMID- 8703884 TI - Retinal haemodynamics in patients with early diabetes mellitus. AB - AIMS/BACKGROUND: The retinal circulation was investigated in a group of 19 patients with insulin dependent diabetes mellitus with less than 4 years of disease duration and no evidence of diabetic retinopathy. Results of these patients were compared with those of 16 age-matched normal controls. METHODS: Venous diameter (D) was measured from monochromatic fundus photographs. Maximum erythrocyte velocity (Vmax) was assessed by bidirectional laser Doppler velocimetry in the major retinal veins of one eye of each subject. Total volumetric blood flow rate (QT) was calculated by adding the flow rates of the major retinal veins. RESULTS: Average QT was 12% larger than normal in diabetic patients (one tailed, non-paired Student's t test, p < 0.05). A statistically significant correlation was observed between QT and disease duration (r = 0.35, p < 0.04). Patients with longer disease duration tended to have somewhat larger QT. The average retinal vascular regulatory responses to hyperoxia were not significantly different from normal in diabetic patients. In these patients, however, higher blood glucose levels were associated with decreased regulatory responses to hyperoxia. CONCLUSIONS: Patients with diabetes mellitus of relatively short duration have mildly increased QT, suggesting that increased blood flow may play an early role in the development of diabetic retinal microangiopathy. PMID- 8703885 TI - Causes and frequency of blindness in patients with intraocular inflammatory disease. AB - AIMS/BACKGROUND: Uveitis, an intraocular inflammatory disease, is a significant cause of visual impairment. It is not known how many patients with uveitis will retain visual acuity and how many develop visual impairment or even blindness. The aim of this study was to assess the frequency of blindness in patients with uveitis and, more specifically, to identify the clinical profile of patients at risk for visual loss. METHODS: A cross sectional and retrospective study of 582 patients with uveitis who visited the ophthalmology departments of two university hospitals in the Netherlands was performed. RESULTS: Within the group of 582 patients, 203 (35%) exhibited blindness or visual impairment; bilateral legal blindness developed in 22 (4%) patients, 26 (4.5%) had one blind eye with visual impairment of the other, and nine (1.5%) had bilateral visual impairment. Unilateral blindness developed in 82 (14%) patients, whereas 64 (11%) exhibited unilateral visual impairment. The most important cause of both blindness and visual impairment was cystoid macular oedema (29% and 41%, respectively). Complications of uveitis were encountered in more than half of the patients and 23% underwent one or more surgical procedures. When the patients were subdivided according to anatomical site, those with panuveitis had the worst visual prognosis. The systemic diseases associated with a poor visual prognosis were juvenile chronic arthritis and sarcoidosis. Ocular toxoplasmosis was the most frequent cause of unilateral visual loss. CONCLUSIONS: Cystoid macular oedema is the most frequent complication of uveitis and its occurrence plays a decisive role in the visual outcome of this disease. PMID- 8703887 TI - Advanced cicatricial retinopathy of prematurity--outcome and complications. AB - AIMS: To assess the outcome and complications of patients with advanced retinopathy of prematurity (ROP). METHODS: All patients with eyes achieving stage 4 or 5 retinopathy of prematurity were reviewed. Twenty one eyes were diagnosed during ROP screening in maternity hospitals and 10 eyes were of infants transferred for treatment. RESULTS: Thirty one eyes of 17 patients were included. Thirteen eyes were treated for acute disease but progressed to stage 4 or 5; seven had cryotherapy and six diode laser photocoagulation. Cataract was found in 17 eyes (54.8%), glaucoma in seven eyes (22.6%), microphthalmos in 15 (48.4%), and corneal opacification in four eyes (12.9%). Fifteen eyes had surgical procedures; two (6.5%) had trabeculectomy, four (12.9%) had lensectomy, and nine (29%) retinal detachment repair. Transferred infants had their initial eye examination later than infants in hospitals screened by the authors and 80% of them had progressed beyond threshold ROP by the time they were transferred for treatment. Twenty nine eyes (93.6%) had visual acuities of 3/60 or less and only two eyes (6.5%) achieved 6/18 or less. CONCLUSION: The visual outcome of the eyes undergoing retinal re-attachment surgery was disappointing. Cataract, microphthalmos, and glaucoma were the most frequent complications, and surgical intervention was often required. The need for children who are blind as a result of ROP to have long term follow up is shown. PMID- 8703886 TI - Onchocerciasis in Ecuador: evolution of chorioretinopathy after amocarzine treatment. AB - AIMS: To investigate the impact of the macrofilaricidal drug, amocarzine, on the evolution of chorioretinopathy in onchocerciasis. METHODS: A prospective uncontrolled cohort study was performed using subjects infected with Onchocerca volvulus in a hyperendemic onchocerciasis focus in Esmeraldas Province in Ecuador. Study subjects were recruited into four cohorts in which ophthalmic and parasitological data were collected for 2, 3, 4, and 5 years respectively. RESULTS: Complete ophthalmic follow up was obtained for 294 individuals in the four cohorts. The incidence of retinal pigment epithelial atrophy tended to remain constant between cohorts while that of chorioretinal scarring with a greater observation period. The incidence rate of cases with new or extending chorioretinal lesions was greater with an increasing period of follow up. An association was seen between the cumulative microfilarial loads in the skin and the development of new chorioretinal lesions (p < 0.05). No relation was noted between cumulative microfilarial loads and the progression of existing disease. CONCLUSION: Amocarzine therapy did not prevent the natural evolution of chorioretinal disease. It was suggested that ocular microfilariae were necessary for the induction of chorioretinopathy in previously unaffected eyes and that extension of existing disease might also be related to the presence of ocular microfilariae or to other immunological mechanisms. PMID- 8703888 TI - Primary vitrectomy for pseudophakic retinal detachment. AB - AIM/BACKGROUND: Viewing the peripheral retina is the major problem in the repair of pseudophakic retinal detachments. Conventional buckling procedures in pseudophakic eyes are complicated by persistent retinal (re-) detachment and proliferative vitreoretinopathy (PVR) more often than in phakic eyes. METHODS: Primary vitrectomy was performed in 33 consecutive cases for pseudophakic retinal detachment with the help of liquid perfluorocarbons and a wide angle viewing system, following a standardised procedure. All eyes have passed the 12 month follow up examination. RESULTS: The primary reattachment rate was 94%. PVR was observed in one case (3%). Seventy nine per cent (26 eyes) regained vision of 20/50 or better, with a median visual acuity of 20/30. The most frequent complication was transient glaucoma during the early postoperative period in 48% (16 eyes) requiring carboanhydrase inhibitors. CONCLUSION: The main advantage of primary vitrectomy over conventional buckling seems to be the better intraoperative sight to the most peripheral retinal holes, controlled removal of vitreous traction, and focused endolaser coagulation. This may explain the low rate of PVR after primary vitrectomy. Also, visual results tended to be better compared with conventional surgical techniques possibly because of removed vitreous opacities, and because of a superior retinal reattachment rate as well as the reduced rate of PVR. PMID- 8703889 TI - Ocular neuromyotonia. AB - AIMS/BACKGROUND: Ocular neuromyotonia is characterised by spontaneous spasm of extraocular muscles and has been described in only 14 patients. Three further cases, two with unique features, are described, and the underlying mechanism reviewed in the light of recent experimental evidence implicating extracellular potassium concentration in causing spontaneous firing in normal and demyelinated axons. METHODS: Two patients had third nerve neuromyotonia, one due to compression by an internal carotid artery aneurysm, which has not been reported previously, while the other followed irradiation of a pituitary tumour, a common association in the published reports. Selective activation occurred in both, where neuromyotonic activity was triggered by prolonged voluntary activation of specific extraocular muscles with or without spread of activity to other third nerve muscles. The other patient had fourth nerve involvement, where spasms of the superior oblique muscle were induced only by alcohol, a phenomenon which has not been described. RESULTS: The two patients with third nerve involvement responded to carbamazepine and in one, an improvement in a chronic partial third nerve paresis occurred. The other has not required treatment and remains asymptomatic by refraining from alcohol. CONCLUSIONS: A careful examination, including the effects of prolonged voluntary muscle action is required to initiate episodes and to demonstrate selective activation. Imaging is mandatory to exclude compressive intracranial lesions, particularly where there is no history of pituitary fossa irradiation. A trial of anticonvulsants should be considered in all patients. Extracellular potassium may play a role in spontaneous firing and ephatic transmission in ocular neuromyotonia. PMID- 8703890 TI - Diffusion coefficient through the blood-aqueous barrier using a standard protocol. AB - AIMS/BACKGROUND: Comparison of the diffusion coefficient through the blood aqueous barrier of healthy volunteers measured in different cities with identical fluorophotometers using a standardised protocol. METHODS: Healthy volunteers aged between 20 and 70 years were studied in seven European cities. The fluorescein concentration in the anterior segment of each eye was measured with a commercial scanning fluorophotometer 30 and 40 minutes after intravenous fluorescein. The decay of non-protein bound fluorescein concentration in blood plasma was determined with the use of three blood samples taken at 7, 15, and 55 minutes after injection. The diffusion coefficient through the blood-aqueous barrier was calculated from the ratio between the fluorescein concentration in the anterior chamber and the time integral of non-protein bound fluorescein concentration in plasma using specially developed software. RESULTS: The mean values of the diffusion coefficient (SD) (X10(-4) min-1) were 4.76 (1.51) (n = 20, Brussels), 5.48 (2.33) (n = 17, Coimbra), 3.47 (2.09) (n = 12, Cologne), 6.09 (2.77) (n = 21, Frankfurt), 3.85 (1.59) (n = 11, Ghent), 4.99 (1.69) (n = 23, Leiden), and 4.87 (1.05) (n = 20, Madrid). The values between centres were similar (Kruskal Wallis test p > 0.05) except for Cologne and Frankfurt (p = 0.013). No differences were found when repeating measurements (four centres, interval time 1 8 months, Wilcoxon paired test p > 0.39). CONCLUSION: The diffusion coefficients had similar values and standard deviations. The concerted action demonstrated the usefulness of a standardised protocol. PMID- 8703891 TI - Levels of vascular endothelial growth factor are elevated in the vitreous of patients with subretinal neovascularisation. AB - BACKGROUND: Vascular endothelial growth factor (VEGF) has been shown to play a major role in intraocular neovascularisation in ischaemic retinal diseases. Subretinal neovascularisation is an important cause of central visual loss, but little is known about the role of this growth factor in its pathogenesis. The aim of this study was to investigate the possible role of VEGF in the development of subretinal neovascularisation. METHODS: Undiluted vitreous samples were obtained from patients undergoing vitrectomy for removal of non-age-related subfoveal neovascular membranes (SFNM). For comparison vitreous from patients undergoing vitrectomy for idiopathic full thickness macular holes (FTMH) and proliferative diabetic retinopathy (PDR) was used. Indirect enzyme linked immunosorbent assay (ELISA), with an antibody directed against the conserved N-terminal region of human VEGF165, was used to determine vitreous levels of VEGF. The growth factor was also localised in the vitreous of patients with SFNM by western blot analysis. RESULTS: The mean (SE) VEGF concentration in the vitreous of patients with SFNM was 27.78 (2.22) ng/ml (n = 8), FTMH was 16.62 (0.9) ng/ml (n = 18), and PDR was 37.77 (3.28) ng/ml (n = 16). The differences between the PDR group and SFNM group versus the FTMH group were both significant (p = 0.0001 and p = 0.0015) as analysed by the Wilcoxon rank sum test). CONCLUSIONS: Vitreous levels of VEGF are significantly elevated in eyes with non-age-related subretinal neovascularisation compared with eyes with FTMH but not as elevated as in PDR. This suggests that VEGF is involved in subretinal angiogenesis. PMID- 8703892 TI - Viability of keratocytes in epikeratophakia lenticules. AB - AIM: To study the influence of cryoprotectant, cooling rate, and warming rate on recovery and viability of keratocytes from corneas for cryolathing. METHODS: Corneas were frozen at -50 degrees C for 2 minutes either after exposure to 10% dimethyl sulphoxide in Eagle's MEM for 15 minutes at room temperature (about 22 degrees C), or without earlier exposure to the cryoprotectant. Corneas were cooled either rapidly (20 degrees C/min) or slowly (1 degree C/min), and they were warmed either rapidly (> 50 degrees C/min) by direct transfer into medium at 22 degrees C or slowly (< 20 degrees C/min) in air at 22 degrees C. The cryoprotectant was removed by dilution in medium containing 0.5 mol/l sucrose. Recovery of keratocytes was determined by using collagenase digestion to release the cells from the stroma and trypan blue staining. Viability was assessed by the outgrowth of cells from stromal explants in primary tissue culture. RESULTS: The use of a cryoprotectant before freezing was beneficial, irrespective of the different cooling and warming regimens. Both collagenase digestion and tissue culture revealed that keratocyte survival was improved when corneas were warmed rapidly rather than slowly. The collagenase digestion assay showed an apparently higher recovery of keratocytes after slow cooling (54.3%) than after rapid cooling (34.1%), but no differences in cell viability could be demonstrated by primary tissue culture. CONCLUSION: Although in these experiments slow cooling apparently provided the best recovery of keratocyte numbers (though not viability), previous work had revealed some disruption of the epithelial basement membrane after slow cooling. If viable keratocytes and good preservation of epithelial basement membrane are considered to be prerequisites for epikeratophakia lenticules then it is suggested that corneas should be prepared for cryolathing by freezing rapidly after exposure to 10% dimethyl sulphoxide and, following cryolathing, they should be warmed rapidly. PMID- 8703893 TI - Physiology of suppression in strabismic amblyopia. PMID- 8703894 TI - Microphthalmos in association with Gorlin's syndrome. PMID- 8703895 TI - Bilateral congenital dentiform cataract and extreme microcornea in eyes with uveal colobomas and persistent hyperplastic primary vitreous. PMID- 8703896 TI - Refractive and visual results and patient satisfaction after excimer laser keratectomy for myopia. PMID- 8703897 TI - When is acute onset concomitant esotropia a sign of serious neurological disease? PMID- 8703898 TI - Low vision. PMID- 8703899 TI - Patient selection may be the key for the success of shared care for glaucoma. PMID- 8703901 TI - Gesnerus index Vol. 1-50, 1943/44-1993. PMID- 8703900 TI - Use of glycopyrrolate as a prophylaxis for vaso-vagal syncope during retinal photocoagulation. PMID- 8703902 TI - Systematic relationship between phospholipase A2 activity and dynamic lipid bilayer microheterogeneity. AB - A standing hypothesis in membrane biology implies that the collective physical properties of the lipid bilayer component of biological membranes can modulate the activity of membrane-associated proteins. We provide strong support for this hypothesis by exploring a model system, phospholipase A2 catalyzed hydrolysis of one-component phospholipid vesicles. For vesicles of lipids with different chain lengths we observe, as a function of temperature and chain length, a systematic variation of the characteristic lag time for the onset of rapid phospholipase A2 hydrolysis. These results, combined with theoretical results obtained from computer simulation of the gel-to-fluid phase transition in the unhydrolyzed lipid bilayers, enable us to demonstrate a strong correlation between the lag time and the degree of bilayer microheterogeneity in the phase transition region. Insight into the nature of this correlation suggests rational ways of modulating enzyme activity by modifying the physical properties of the lipid bilayer. PMID- 8703903 TI - Anion binding by transferrins: importance of second-shell effects revealed by the crystal structure of oxalate-substituted diferric lactoferrin. AB - Proteins of the transferrin family bind, with high affinity, two Fe3+ ions and two CO3(2)- ions but can also bind other metal ions and other anions. In order to find out how the protein structure and its two binding sites adapt to the binding of larger anions, we have determined the crystal structure of oxalate-substituted diferric lactoferrin at 2.4 A resolution. The final model has a crystallographic R-factor of 0.196 for all data in the range 8.0-2.4 A. Substitution of oxalate for carbonate does not produce any significant change in the polypeptide folding or domain closure. Both binding sites are perturbed, however, and the effects are different in each. In the C-lobe site the oxalate ion is bound to iron in symmetric 1,2-bidentate fashion whereas in the N-lobe the anion coordination is markedly asymmetric. The difference arises because in each site substitution of the larger anion causes displacement of the arginine that forms one wall of the anion binding site; the movement is different in each case, however, because of different interactions with "second shell" amino acid residues in the binding cleft. These observations provide an explanation for the site inequivalences that accompany the substitution of non-native anions and cations. PMID- 8703904 TI - Global mapping of structural solutions provided by the extended X-ray absorption fine structure ab initio code FEFF 6.01: structure of the cryogenic photoproduct of the myoglobin-carbon monoxide complex. AB - X-ray methods based on synchrotron technology have the promise of providing time resolved structural data based on the high flux and brightness of the X-ray beams. One of the most closely examined problems in this area of time-resolved structure determination has been the examination of intermediates in ligand binding to myoglobin. Recent crystallographic experiments using synchrotron radiation have identified the protein tertiary and heme structural changes that occur upon photolysis of the myoglobin--carbon monoxide complex at cryogenic temperatures [Schlichting, I., Berendzen, J., Phillips, G., & Sweet, R. (1994) Nature 371, 808--812]. However, the precision of protein crystallographic data (approximately 0.2 A) is insufficient to provide precise metrical details of the iron--ligand bond lengths. Since bond length changes on this scale can trigger reactivity changes of several orders of magnitude, such detail is critical to a full understanding of metalloprotein structure--function relationships. Extended X-ray absorption fine structure (EXAFS) spectroscopy has the potential for analyzing bond distances to a precision of 0.02 A but is hampered by its relative insensitivity to the geometry of the backscattering atoms. Thus, it is often unable to provide a unique solution to the structure without ancillary structural information. We have developed a suite of computer programs that incorporate this ancillary structural information and compute the expected experimental spectra for a wide ranging series of Cartesian coordinate sets (global mapping). The programs systematically increment the distance of the metal to various coordinating ligands (along with their associated higher shells). Then, utilizing the ab initio EXAFS code FEFF 6.01, simulated spectra are generated and compared to the actual experimental spectra, and the differences are computed. Finally, the results for hundreds of simulations can be displayed (and compared) in a single plot. The power of this approach is demonstrated in the examination of high signal to noise EXAFS data from a photolyzed solution sample of the myoglobin--carbon monoxide complex at 10 K. Evaluation of these data using our global mapping procedures placed the iron to pyrrole nitrogen average distances close to the value for deoxymyoglobin (2.05 +/- 0.01 A), while the distance from iron to the proximal histidine nitrogen is seen to be 2.20 +/- 0.04 A. It is also shown that one cannot uniquely position the CO ligand on the basis of the EXAFS data alone, as a number of reasonable minima (from the perspective of the EXAFS) are observed. This provides a reasonable explanation for the multiplicity of solutions that have been previously reported. The results presented here are seen to be in complete agreement with the crystallographic results of Schlichting et al. (1994) within the respective errors of the two techniques; however, the extended X-ray absorption fine structure data allow the iron--ligand bond lengths to be precisely defined. An examination of the available spectroscopic data, including EXAFS, shows that the crystallographic results of Schlichting et al. (1994) are highly relevant to the physiological solution state and must be taken into account in any attempt to understand the incomplete relaxation process of the heme iron for the Mb*CO photoproduct at low temperature. PMID- 8703905 TI - Involvement of arginine 143 in nucleotide substrate binding at the active site of adenylosuccinate synthetase from Escherichia coli. AB - Adenylosuccinate synthetase from Escherichia coli is inactivated in a biphasic reaction by guanosine 5'-O-[S-(4-bromo-2,3-dioxobutyl)thio]phosphate (GMPSBDB) at pH 7.1 and 25 degrees C. Reaction of the enzyme with [8-3H]GMPSBDB results in the incorporation of 2 mol of the reagent/mol of subunit; in the presence of active site ligands the incorporation is reduced to 1 mol of reagent/mol of subunit. GMPSBDB reacts with Cys-291 in the initial rapid reaction which is accompanied by loss of 50% of the enzymatic activity; this reaction is not affected by the presence of active site ligands. In the slower reaction, GMPSBDB inactivates the enzyme by reacting with Arg-143. The inactivation kinetics of the slower phase are consistent with the formation of an enzyme--GMPSBDB complex having a Kd of 42 microM. Active site nucleotides, either adenylosuccinate or IMP + GTP, prevent both slower phase inactivation and labeling of Arg-143. Replacement of Arg-143 with a Leu by site-directed mutagenesis does not change the catalytic constant or the K(m) for aspartate but does significantly impair nucleotide binding: the Michaelis constants for IMP and GTP increase by 60-fold and 10-fold, respectively, in the R143L mutant. The crystal structure of the E. coli enzyme [Poland, B.W., Silva, M.M., Serra, M.A., Cho, Y., Kim, K. H., Harris, E.M.S., & Honzatko, R.B. (1993) J. Biol. Chem. 268, 25334--25342] shows that Arg-143 from one subunit projects into the putative active site of the other subunit. These results indicate that both subunits of dimeric adenylosuccinate synthetase contribute to each active site and that Arg-143 plays an important role in nucleotide binding. PMID- 8703906 TI - Identification of residues critical to the activity of human granulocyte colony stimulating factor. AB - Alanine scanning mutagenesis of human granulocyte colony-stimulating factor (G CSF) was used to identify residues critical for the cell-proliferative activity of the protein. Fifty-eight residues, most of them on the protein surface, were independently mutated to alanine. Most of the variants retained full biological activity; however, 15 mutants were significantly impaired in their ability to stimulate bone marrow cell proliferation in vitro. Four of these variants contain mutations at buried residues and two have substitutions at side chains involved in intramolecular hydrogen bonds. The remaining nine down mutations identify two regions on the surface of the molecule important for biological activity. Consistent with these observations, measurements of binding to NFS-60 cells indicate that the residues most important for receptor binding are Lys40 and Phe144 in site 1 and Glu19 in site 2. In addition to these residues, Val48 and Leu49 in site 1 and Leu15, Asp112, and Leu124 in site 2 are also important for biological activity. These results suggest the presence of two binding sites on the cytokine surface required for dimerization of the G-CSF receptor. PMID- 8703907 TI - In vitro enzyme activation and folded stability of Pseudomonas aeruginosa exotoxin A and its C-terminal peptide. AB - Pseudomonas aeruginosa exotoxin A(ETA) and its C-terminal, enzymatically active fragment (PE40, 375 residues) were studied by high-performance size-exclusion chromatography, steady-state and stopped-flow fluorescence spectroscopy, and circular dichroism spectroscopy. Both proteins have been overexpressed and purified by high-performance liquid chromatography. The effect of various activation conditions (pH, urea, and DTT) on enzymatic activity was studied. Upon enzymatic activation, structural changes induced within both proteins' structures were monitored, and these changes were correlated with concomitant alterations in the catalytic activity of the proteins. The pH optimum of enzymatic activity for both ETA and PE40 was between 7.0 and 8.0, decreasing to nearly zero at acidic (pH 5.0) and basic (pH 11-12) values. Analysis of the pH titration data revealed the presence of two distinct pKa values which implicate a His residue(s) (likely His-440 and -426) and a Tyr or Lys residue (possibly Tyr-481). The identity and possible role of an active site Lys residue is not known. Additionally, a significant increase in the Stokes radii of both proteins was detected when the pH was lowered from 8.0 to 6.0. The enzymatic activity of PE40 was not affected by urea or DTT, and its Stokes radius decreased monotonically with increasing urea concentration in the presence of DTT. In contrast, the enzymatic activity of ETA peaked when the protein was preincubated with 4.0 M urea, and this coincided with a large transition (increase) in the protein's Stokes radius between 3 and 5 M urea. Furthermore, loss of helical secondary structure of both PE40 and ETA commenced at approximately 2 M urea and progressively diminished at higher denaturant concentrations. The unfolding of both proteins in urea (and DTT) was reversible, and the free energies of unfolding were determined by both circular dichroism and fluorescence spectroscopy and were found to be 13.7 +/- 2.9 and 9.8 +/- 3.4 kJ/mol, respectively, for ETA and were 17.8 +/- 6.8 and 7.5 +/- 3.6 kJ/mol, respectively, for PE40. The refolding rate of PE40 was relatively rapid [t 1/2(1) = 27 s, t 1/2(2) = 624 s], which was in stark contrast to the refolding rate of ETA (t 1/2 = several hours). The relative refolding rates of PE40 and ETA help to explain the mechanism of in vitro enzyme activation and assay. PMID- 8703908 TI - Steroidogenic factor 1-dependent promoter activity of the human steroidogenic acute regulatory protein (StAR) gene. AB - Steroidogenic acute regulatory protein (StAR) is required for efficient adrenal cortical and gonadal but not trophoblast steroid hormone synthesis. StAR gene expression in gonadal cells is stimulated by tropic hormones acting through the intermediacy of cAMP. DNA sequence analysis of the human StAR gene promoter revealed two motifs resembling binding sites for steroidogenic factor 1 (SF-1), a member of the orphan nuclear receptor transcription factor family that controls expression of steroidogenic hydroxylases. The 5'-most sequence (distal site) is a consensus SF-1 binding site. The proximal site is a consensus estrogen receptor binding half-site. The StAR gene promoter is not active in BeWo choriocarcinoma cells, COS-1 cells, HeLa cells, or SK-OV-3 ovarian adenocarcinoma cells, all of which do not express significant levels of SF-1 mRNA. Introduction of SF-1 into these cells stimulated StAR promoter activity, particularly in response to cAMP. Two orphan nuclear transcription factors that bind to sequences similar to SF-1 sites, NGFI-B/Nur77 and RNR-1, did not support cAMP-stimulated StAR promoter activity in BeWo cells. Mutation of the distal putative SF-1 binding site reduced basal and cAMP-stimulated promoter activity in BeWo cells by 82% and 71%, respectively. Mutation of the proximal putative SF-1 binding site reduced basal and cAMP-stimulated promoter activity by 89% and 96%, respectively. Mutations in both sites reduced basal promoter activity to 7% of wild type promoter activity and cAMP-stimulated promoter activity to less than 5% of the wild type. Deletion analyses of promoter activity were consistent with the mutation studies. Electrophoretic mobility shift assays (EMSAs) demonstrated that the distal site binds to SF-1 expressed in COS-1 cells and to an SF-1-GST fusion protein with high affinity, but that the mutated distal sequence does not. An anti-SF-1 antibody ablated the characteristic SF-1-DNA complex with the distal sequence. The proximal site formed a number of protein-DNA complexes with COS-1 cell extracts, but appeared to have at best only very modest affinity for SF-1. Collectively, our findings demonstrate that SF-1 plays a key role in controlling the basal and cAMP-stimulated expression of the StAR gene. SF-1 can function at two distinct sites in the human StAR gene promoter, apparently by two different types of interaction, to control transcription. PMID- 8703909 TI - Regulation of human C-reactive protein gene expression by two synergistic IL-6 responsive elements. AB - To study the mechanism of interleukin-6 (IL-6) induction of human C-reactive protein (CRP) gene expression, we have utilized a human hepatoma (PLC/PRF/5) cell culture system to analyze the trans-acting factors which bind to the 300 bp 5' flanking region of human CRP gene. In vitro gel mobility shift analyses and methylation interference assays demonstrated that NFIL-6 alpha interacted with two IL-6 responsive elements, and HNF-1 alpha and HNF-3/Octamer-like factors interacted with the downstream IL-6 responsive element in the human CRP promoter. In vivo functional analysis by transient transfection of plasmid constructs containing site-specific mutations in one or two IL-6 responsive elements in the CRP promoter fused to a reporter gene, chloramphenicol acetyl transferase (CAT), demonstrated that the binding of NFIL-6 alpha to two IL-6 responsive elements resulted in synergistic induction of the gene. When HNF-1 alpha or HNF-3/Octamer like factors were independently bound to their corresponding sites, they had either a positive or negative effect, respectively, on IL-6 inducible transcriptional activity. PMID- 8703910 TI - Development of a sensitive peptide-based immunoassay: application to detection of the Jun and Fos oncoproteins. AB - c-Jun and c-Fos belong to the bZIP class of transcriptional activator proteins, many of which have been implicated in the neoplastic transformation of cells. We are interested in engineering dominant-negative leucine zipper (LZ) peptides as a means of sequestering these proteins in vivo in order to suppress their transcriptional regulatory activity. Toward this end, we have developed a novel immunoassay for measuring the dimerization affinities of dimeric Jun and Fos complexes. This peptide-based ELISA relies on the fact that Jun and Fos preferentially form heterodimers via their leucine zipper domains. Recombinant Jun leucine zipper peptides (either native JunLZ or a V36 --> E point mutant) were labeled with biotin and specifically bound through a leucine zipper interaction to a FosLZ-glutathione S-transferase fusion protein adsorbed onto the wells of an ELISA tray. Jun:Fos complexes were subsequently detected using a recently developed streptavidin-based amplification system known as enzyme complex amplification [Wilson, M. R., & Easterbrook-Smith, S.B. (1993) Anal. Biochem. 209, 183-187]. This ELISA system can detect subnanomolar concentrations of Jun and Fos, thus allowing determination of the dissociation constants for complex formation. The dissociation constant for formation of the native JunLZ:FosLZ heterodimer at 37 degrees C was determined to be 0.99 +/- 0.30 nM, while that for JunLZ(V36E):FosLZ heterodimer was 0.90 +/- 0.13 microM. These results demonstrate that the novel peptide-based ELISA described herein is simple and sensitive and can be used to rapidly screen for potential dominant-negative leucine zipper peptides. PMID- 8703911 TI - Kinetics of the interaction of nonsteroidal antiinflammatory drugs with prostaglandin endoperoxide synthase-1 studied by limited proteolysis. AB - Many nonsteroidal antiinflammatory agents (NSAIDs) bind to prostaglandin endoperoxide synthase (PGHS) and induce a conformational change in the PGHS apoprotein that renders it resistant to cleavage by trypsin at Arg277. In the present study, the trypsin protection assay was modified to permit detection of conformational changes at times as short as 5 s after the addition of inhibitor. The kinetics of the induction and reversal of trypsin resistance in apoPGHS-1 by a series of NSAIDs and isozyme-specific PGHS-1 and PGHS-2 inhibitors were determined. All compounds induced resistance to trypsin cleavage at a rapid rate. The conformational change induced by competitive inhibitors was reversed on prolonged incubation with trypsin (approximately 5 min). In contrast, the resistance induced by irreversible inhibitors was not lost during a 5 min incubation with trypsin. All of the selective PGHS-2 inhibitors protected against tryptic cleavage of apoPGHS-1 but did not inhibit the protein's cyclooxygenase activity. The results suggest that induction of trypsin resistance is a reflection of the initial association of reversible as well as irreversible inhibitors with the apoprotein. PMID- 8703912 TI - Bothrojaracin: a potent two-site-directed thrombin inhibitor. AB - The thrombin inhibitor, bothrojaracin [Zingali, R. B., Jandrot-Perrus, M., Guillin, M. C., & Bon, C. (1993) Biochemistry 32, 10794-10802], is a 27 kDa protein isolated from the venom of Bothrops jararaca that blocks several thrombin functions, including fibrinogen clotting, platelet activation, and fibrin and thrombomodulin binding, but does not interact with the catalytic site. In the present report, we show that the high affinity binding of alpha-thrombin to immobilized bothrojaracin (Kd = 0.6 nM) is inhibited by the C-terminal peptide of hirudin and that the gamma-cleavage within exosite 1 reduces the affinity of bothrojaracin for thrombin (Kd = 0.3 microM), indicating that bothrojaracin binding to exosite 1 is a major determinant of the thrombin-bothrojaracin interaction. In addition, we show that bothrojaracin decreases the rate of inhibition of alpha- and gamma-thrombin by the antithrombin III-heparin complex. Competition of bothrojaracin with heparin or prothrombin fragment 2 for binding to thrombin indicates that bothrojaracin not only binds exosite 1 but also binds exosite 2 or in close proximity. Bothrojaracin binds to the thrombin precursor, prothrombin. This interaction is calcium-independent and is prevented by heparin, suggesting that it is mediated by exosite 2. Bothrojaracin inhibits platelet activation induced by clot-bound thrombin and slowly dissociates thrombin from the fibrin clots. Altogether, our results indicate that the high affinity of bothrojaracin for thrombin is supported by a double-site interaction and results in an efficient inhibition of both soluble and clot-bound thrombin. PMID- 8703913 TI - Inhibition of human leukocyte and porcine pancreatic elastase by homologues of bovine pancreatic trypsin inhibitor. AB - The interactions of three BPTI homologues with human leukocyte elastase and porcine pancreatic elastase have been investigated. The principal mutation in determining the specificity of inhibition was the Lys15-Val mutation at the P1 position. An additional mutation at P3, i.e., BPTI (Lys15-Val, Pro13-Ile), increased the inhibition of HLE to a Ki = 2.5 x 10(-10) M, but decreased the inhibition of PPE, showing this to be a useful site for improving selectivity. Kinetic evidence suggests that the inhibition of HLE by BPTI homologues probably takes place by a two-step mechanism in which an isomerization step occurs after initial binding. 1H NMR spectroscopy of the BPTI (Lys15-Val) and BPTI (Lys15-Val, Pro13-Ile) mutants indicates that small conformational changes are associated with the mutations, but these are localized in the immediate vicinity of the mutation in the outer binding loop and in the inner loop connected to it through the Cys14-Cys38 disulfide bridge. PMID- 8703914 TI - Chemical modification of recombinant HIV-1 capsid protein p24 leads to the release of a hidden epitope prior to changes of the overall folding of the protein. AB - It was found that the affinity of a monoclonal antibody directed against a recombinantly expressed HIV-1 capsid protein p24 (rp24) strongly increased after chemical modification of the Iysine residues of rp24 with different amounts of maleic anhydride. The extent and the sites of modification were analyzed by MALDI TOF mass spectrometry. Unmodified rp24 and the differently modified rp24 samples were tested for binding the murine monoclonal antibody CB4-1 which recognizes the epitope GATPQDLNTML comprising residues 46-56 of rp24. An increase in the number of modified lysine residues led to enhanced binding affinity of CB4-1. Most pronounced effects were observed after substitution of the first amino groups: an average number of three modified residues per protein molecule increases the binding affinity by a factor of 23, but the substitution of the remaining nine residues increases the binding affinity only by a factor of 11. Fully modified rp24 variant proteins were bound by CB4-1 with Kd values comparable to that of the peptide epitope. Conformation and stability of the unmodified rp24, highly (rp24F, 9 residues; rp24G, 11 residues) modified, and fully modified protein (rp24I, 11 lysine residues and N-terminus) were analyzed by circular dichroism (CD) and fluorescence spectroscopy under different solvent conditions. Little difference in conformation and unfolding behavior was observed between the unmodified and highly modified rp24, which differ drastically in the antibody binding behavior. The fully modified sample, however, displayed a significant decrease in alpha-helical content. Thus, the epitope seems to be hidden (cryptotope) in the unmodified rp24 in a low-affinity binding conformation and becomes displayed at low levels of chemical modification which obviously induce subtle structural changes prior to changes of the overall folding observable by spectroscopic means. PMID- 8703915 TI - Recombinant toluene-4-monooxygenase: catalytic and Mossbauer studies of the purified diiron and rieske components of a four-protein complex. AB - Expression of the tmoA-F gene cluster from Pseudomonas mendocina KRI in Escherichia coli BL21(DE3) produces a catalytically active form of the toluene-4 monooxygenase (T4MO) complex. Here we report the purification and characterization of four soluble proteins required for the in vitro reconstitution of T4MO catalytic activity. These proteins are a diiron hydroxylase (T4MOH), a Riesketype ferredoxin (T4MOC), an effector protein (T4MOD), and an NADH oxidoreductase (T4MOF). The T4MOH component is composed of the tmoA, tmoB, and tmoE gene products [quaternary structure (alpha beta epsilon)2, Mr approximately 220 kDa]. The T4MOA polypeptide contains two copies of the amino acid sequence motif (D/E)X(28-37)DEXRH; the same motif provides all of the protein-derived ligands to the diiron centers of ribonucleotide reductase, the soluble methane monooxygenase, and the stearoyl-ACP delta 9 desaturase. Mossbauer, optical, and EPR measurements show that the T4MOH contains diiron centers and suggest that the diiron center contains hydroxo bridge(s) in the diferric state, as observed for methane monooxygenase. Mossbauer and EPR measurements also show that the T4MOC contains a Rieske-type iron-sulfur center. This assignment is in accord with the presence of the amino acid sequence motif CPHX(15-17)CX2H, which has also been found in the bacterial, chloroplastic, and mitochondrial Rieske proteins as well as the bacterial NADH-dependent cis dihydrodiol-forming aromatic dioxygenases. While single-turnover catalytic studies confirm the function of the T4MOH as the hydroxylase, the NADH-dependent multiple-turnover hydroxylation activity is increased by more than 100-fold in the presence of the T4MOC, which mediates highly specific electron transfer between the T4MOF and the T4MOH. The T4MOD can be purified as an 11.6 kDa monomeric protein devoid of cofactors or redox-active metal ions; this component is also detected as a substoichiometric consitutent of the purified T4MOH. The rate of the hydroxylation reaction can be mildly stimulated by the further addition of separately purified T4MOD to the T4MOH, implying the formation of a high affinity, catalytically competent complex between these two components. These characterizations define a novel, four-component oxygenase combining elements from the soluble methane oxidation complex of the methanotrophic bacteria and the aromatic hydroxylation complexes of the soil pseudomonads. PMID- 8703916 TI - Structural studies of the proton-translocating NADH-quinone oxidoreductase (NDH 1) of Paracoccus denitrificans: identity, property, and stoichiometry of the peripheral subunits. AB - The proton-translocating NADH-quinone oxidoreductase (NDH-1) of Paracoccus denitrificans is composed of at least 14 unlike subunits and contains one FMN and at least five EPR-detectable iron-sulfur clusters. The 14 subunits are designated NQO1 through NQO14. The expression and partial characterization of the NQO4, -5, and -6 subunits have been performed. The NQO4, -5, and -6 subunits were individually expressed in Escherichia coli. The NQO4 subunit was expressed in both the cytoplasmic phase and membrane fraction, the NQO5 subunit in the cytoplasmic phase only, and the NQO6 subunit in the membrane fraction only. The NQO4 and NQO5 subunits were purified from cytoplasmic phase. Neither subunit contains non-heme iron or acid-labile sulfide, suggesting that the NQO4 or NQO5 subunit is not an iron-sulfur subunit. The antibodies against the NQO4, -5, and 6 subunits cross-reacted with their counterpart subunits in bovine heart complex I. The NQO4, -5, and -6 subunits in membrane-bound P. denitrificans NDH-1 were extracted by treatment at alkaline pH ( > or = 10) or with chaotropes (NaBr, Nal, and urea), suggesting that these subunits are localized in the peripheral part (not in the membrane sector) of the enzyme complex similar to the NQO1, -2, and 3 subunits. In addition, the subunit stoichiometry of NQO1 through -6 of the membrane-bound P. denitrificans NDH-1 has been determined by radioimmunoassays. There is 1 mol each of the NQO1 through -6 subunits per mol of the P. denitrificans NDH-1. PMID- 8703917 TI - Structural determinants of the uridine-preferring specificity of RNase PL3. AB - RNase PL3 is a structurally highly conserved, pyrimidine-specific RNase, which strongly prefers to cleave at the 3'-side of uridine. Here, question of which residues are involved in determining substrate specificity is addressed. The difference in the rate of cleavage of UpA and CpA was found to result from a 375 fold larger kcat for the former substrate, whereas the values of Km were essentially the same. The pyrimidine specificity of this class of RNases is thought to result from hydrogen bonds between the base and a threonine residue in the B1 subsite. Mutation of this residue (Thr-44) in RNase PL3 resulted in strongly reduced activity with UpA and poly(U). However, the activity with CpA and poly(C) had increased. Comparison with the effect of the same mutation in RNase A [delCardayre, S. B., & Raines, R. T. (1994) Biochemistry 33, 6031-6037] and angiogenin [Curran et al. (1993) Biochemistry 32, 2307-2313] showed that the function of this threonine in substrate recognition is different in three RNase subfamilies. Previous studies have shown that the 36-42 region contains one or more residues that are involved in substrate recognition [Vicentini et al. (1994) Protein Sci. 3, 459-466]. Site-directed mutagenesis of amino acids in this region identified Phe-42 as the only single residue that affected the cytidine/uridine specificity ratio. The mutation F42V resulted in a 10-fold increase in kcat and a 1.9-fold decrease in Km for CpA. The properties of the double mutant F42V/T44A suggested that a suboptimal binding of cytidine is caused by Phe-42, partially through an effect on Thr-44. PMID- 8703918 TI - Distamycin A complexation with a nucleic acid triple helix. AB - The interaction of the minor groove binding drug distamycin with the T-A-T triple helix and the A-T double helix was studied using circular dichroism spectroscopy and thermal denaturation. The triple helix was made by the oligonucleotide (dA)12 x-(dT)12-x-(dT)12, where x is a hexaethylene glycol chain bridged between the 3' phosphate of one strand and the 5'-phosphate of the following strand. This oligonucleotide is able to fold back on itself to form a very stable triplex. Changing the conditions allows the same oligonucleotide to be in a duplex form with a dangling arm. Circular dichroism spectroscopy demonstrates that the distamycin A molecule can bind to the triple-stranded form of this oligonucleotide. Spectral analysis shows that the bound distamycin exhibits a conformation and an environment slightly different from those which are observed when the drug is bound to the corresponding double-stranded structure. Furthermore, a second type of complex which is observed in the double-strand binding (two stacked distamycins in the minor groove) is not observed with the triple-stranded host. When distamycin is added to the triplex made of unbridged chains (dA)12 + 2(dT)12, the triplex dissociates to give a double-stranded structure. Thermal denaturation experiments demonstrate that distamycin binding destabilizes the triplex whereas it stabilizes the duplex. These results are compared with those obtained by the same experimental approaches on other minor groove binding drugs. PMID- 8703919 TI - Structure/nuclease activity relationships of DNA cleavers based on cationic metalloporphyrin-oligonucleotide conjugates. AB - The covalent attachment of a managanese-tris(methylpyridiniumyl)porphyrin entity to an antisense oligonucleotide allowed sequence-selective oxidative cleavage of DNA when the metalloporphyrin was activated by potassium monopersulfate (KHSO5). We prepared several structurally modified metallo-porphyrin-oligonucleotide conjugates in order to find out the most efficient compound for in vitro DNA cleavage. The nature and the length of the tether were modulated, the metalloporphyrin entity was modified (metal, ligand), and different ways of activation of the metalloporphyrin were assayed. We noticed that the location of the peptidic bond within the linker could greatly affect the cleavage efficiency of the different conjugates. We showed that the most efficient conjugate for oxidative DNA cleavage was a manganese tetracationic porphyrin-oligonucleotide compound. When the metalloporphyrin moiety was activated by a reducing agent in the presence of molecular oxygen, DNA cleavage was efficient at suitable concentrations of the reducing agent, in order to avoid the reduction of the activated DNA cleaver, a putative high-valent metal-oxo species, by the excess of reducing agent. PMID- 8703920 TI - Motional dynamics of a buried tryptophan reveals the presence of partially structured forms during denaturation of barstar. AB - A double mutant of the single-domain protein barstar having a single tryptophan (W53) was made by mutating the remaining two tryptophans (W38 and W44) into phenylalanines. W53 is buried in the core of barstar. Time-resolved fluorescence of the mutant barstar (W38FW44F) showed that W53 has a single fluorescence lifetime in the native (N) state and has three lifetimes in the molten globule like low-pH (A) form. Quenching of fluorescence by either KI or acrylamide showed that W53 is solvent inaccessible in the N-state and fairly accessible in the A form. The denaturation of W38FW44F by guanidine hydrochloride (GdnHCI) was monitored by several probes: near-UV and far-UV circular dichroism (CD), fluorescence intensity, and steady-state and time-resolved fluorescence anisotropy. While the unfolding transitions observed through CD and fluorescence intensity coincided with each other (midpoint approximately 1.8 M GdnHCI), the transition observed through the steady-state fluorescence anisotropy was markedly different from others. Initially, the anisotropy increased with the increase in the concentration of GdnHCI and decreased subsequently. The midpoint of this titration was 2.2 M GdnHCI. Picosecond time-resolved fluorescence anisotropy showed that W38FW44F has a single rotational correlation time of 4.1 ns in the native (N) state and 1.5 as in the unfoled (U) state (6 M GdnHCI). These could be explained as being due to the absence of motional freedom of W53 in the N-state and the presence of rotational freedom in the U-state. In the intermediate concentration region (1.8-3.0 M GdnHCI), the anisotropy decays showed at least two coorrelation times, approximately 1 and 6-12 ns. These two correlation times are ascribed to partially structured forms leading to hindered rotation of W53. Thus, the usefulness of time-resolved fluorescence anisotropy in detecting partially folded structures is demonstrated. PMID- 8703921 TI - Domain organization and DNA-induced conformational changes of an archaeal family B DNA polymerase. AB - Family B DNA polymerase from the thermoacidophilic archaeon Sulfolobus solfataricus (Sso DNA pol) is a monomer of about 100 kDa with two associated catalytic functions: 3'-5' exonuclease and DNA polymerase activities. The structure of this enzyme in the free and DNA-bound states was probed by limited proteolysis and fluorescence spectroscopy measurements. The results of partial trypsin proteolysis experiments on the recombinant Sso DNA pol pinpointed three major sites of protease sensitivity: near the N-terminus, within the center, and near the C-terminal end of the polypeptide chain. When partial trypsin digestion was carried out in the presence of either activated calf thymus DNA or primed M13mp18 single-stranded DNA, changes in cleavage pattern and in susceptibility to protease were detected. This phenomenon was dependent on the nucleic acid concentration and suggested the occurrence of DNA-induced conformational changes. These were also probed by steady-state fluorescence spectroscopy measurements using acrylamide as a quencher. Fine mapping of the DNA-specific cleavage sites allowed us to precisely locate the protein subdomains which were affected by these structural changes. Importantly, a specific proteolytic fragment of about 8 kDa was recovered after partial digestion of Sso DNA pol only in the presence of nucleic acid ligands. It was found to start at residues 392-394 and to span the protease-hypersensitive central region of the polypeptide chain. Its involvement in critical polymerase functions, such as substrate binding and/or enzyme processivity, was discussed. In addition, we found that controlled trypsin digestion of Sso DNA pol did not inactivate either polymerase or 3'-5' exonuclease activity concomitantly with the disappearance of full-sized enzyme. Activity gel analysis revealed that proteolytic products corresponding to the amino- and carboxyl-terminal halves of the enzyme retained 3'-5' exonuclease and DNA polymerase activity, respectively. These results are in line with the model of modular organization proposed for Sso DNA pol in a previous report. PMID- 8703922 TI - Conformational change and inactivation of membrane phospholipid-related activity of cardiotoxin V from Taiwan cobra venom at acidic pH. AB - The phospholipid binding activity of cardiotoxin V from Naja naja atra (CTX A5) was studied by use of Langmuir monolayers and found to exhibit pH-dependence in binding to phosphatidylcholine membrane with an apparent pKa around 6.0. Proton NMR investigation of the CTX A5 molecule in the presence of phosphatidylcholine micelles reveals a decrease in association of CTX A5 with membranes at low pH as a result of the protonation of His-4 near the membrane binding site of loop I region of CTX. The pH-dependent binding can be attributed mainly, but not solely, to the change in charge content of the CTX molecule upon His-4 protonation at the membrane/water interface. This is shown by analyzing the pH- and ionic strength dependence of binding of CTXs to phospholipid monolayers according to Gouy Chapman theory. The protonation of the His-4 residue also results in a local conformational change in the loop I region since the chemical shifts of amide protons for the amino acid residues from Cys-3 to Thr-14 are all found to vary as a function of pH with an apparent pKa similar to that of His-4. Interestingly, the effect is relayed to other amino acid residues in the structural core of the protein such as those in C-terminal (Lys-60, Cys-61, and Asn-62) and triple stranded antiparallel beta-sheet (Cys-22, Lys-24, Ala-25, Arg-38, and Ala-41) regions. An additional local conformational change in the molecule results around pH 5 as evidenced by circular dichroism spectroscopic studies, although this change does not affect the characteristic beta-sheet and three-finger loop structure of CTX molecule as revealed by two-dimensional NOESY 1H NMR study. The latter conformational change at acidic pH, however, completely inactivates CTX induced aggregation/fusion activity of sphingomyelin vesicles. The results suggest that deciphering the functional sites of CTXs on the basis of structure and dynamics determined at low pH should be done with caution. Since 19 out of 44 CTX homologues with known amino acid sequence contain His-4, the effect of His-4 on the structure and function of CTX molecules is important and is discussed in terms of the diverse membrane targets of CTX subtypes. Also discussed is the pH induced activation of snake venom proteins in the victim. PMID- 8703923 TI - The role of acidic amino acid residues in the structural stability of snake cardiotoxins. AB - We have recently shown that membrane-related activities of cardiotoxin V from Naja naja atra (CTX A5) are diminished at acidic pH although the overall beta sheet structure of the molecule is maintained. In order to understand more about the mechanism of inactivation of CTX at acidic pH, we studied the effect of pH and denaturing reagents on the structural stability of CTX. We found, first, pH induced structural transitions occurred in CTX A5 at two pH values as judged by the CD ellipticity around 195 nm: an increase in the beta-sheet content occurred around pH 4 and followed by a decrease, therein, around pH 2. The pKa of three acidic amino acid residues in CTX A5, i.e., Glu-17, Asp-42, and Asp-59, were determined to be 4.0, 3.2, and below 2.3, respectively, by NMR spectroscopy. The low pKa value of Asp-59 implies salt bridge formation between Lys-2 and Asp-59. Thus, electrostatic interaction may stabilize the three loop structure in addition to the hydrogen bonds between N- and C-termini of CTX molecule. Second, 2,2,2-trifluoroethanol (TFE) and guanidinium chloride (GdmHCI) were found to induce alpha-helical and random coil formation, respectively, in CTX A5 and eight other beta-sheet CTXs. Comparison of the relative potencies of TFE and GdmHCI to induce structural changes suggests that the amino acid residue located at position 17 plays a role in the structural stability. Specifically, CTXs containing negatively charged Glu-17 are least stable. It is suggested that Glu 17 may perturb the interaction between Lys-2 and Asp-59, and thus the overall stability of beta-sheet, in the presence of denaturing reagent. In conclusion, the perturbed structural stability of CTXs may partially explain the lower activity CTX exhibits at acidic pH. A structural model to account for the unfolding and refolding of CTX molecules without the breaking of disulfide bonds is also proposed. PMID- 8703924 TI - Relationship between arachidonate--phospholipid remodeling and apoptosis. AB - Our previous studies reveal that three structurally distinct inhibitors of the enzyme CoA-independent transacylase, including the antiproliferative alkyllysophospholipid ET-18-O-CH3, induce programmed cell death (apoptosis) in the promyelocytic cell line HL-60. The objective of the current study was to better elucidate the mechanism responsible for apoptosis. CoA-IT is an enzyme believed to be responsible for the remodeling of long chain polyunsaturated fatty acids like arachidonate between the phospholipids of mammalian cells. The chronic (24-48 h) treatment of HL-60 cells with all three CoA-IT inhibitors resulted in the inhibition of the remodeling of labeled arachidonate from choline- into ethanolamine-containing phospholipid molecular species. GC-MS analysis of the fatty acids in phospholipids revealed that CoA-IT inhibitor treatment induced a marked loss of arachidonate-containing phosphatidylethanolamine and an increase in arachidonate-containing phosphatidylcholine. This redistribution was specific to arachidonate since the mass distribution of linoleic acid in glycerolipids was not affected. In spite of the dramatic redistribution of arachidonate, the total cellular arachidonate content was not altered nor was the relative distribution of total phospholipid classes. The increase of arachidonate in phosphatidylcholine was specifically due to an increase in 1-acyl-2-arachidonoyl sn-glycero-3-phosphocholine species, whereas the loss of arachidonate in PE was from both 1-acyl- and 1-alk-1-enyl-2-arachidonoyl-sn-glycero-3 phosphoethanolamine species. The incubation of cells with exogenous arachidonic acid or ethanolamine did not reverse the inhibition of proliferation induced by CoA-IT inhibitor treatment. Incubation with CoA-IT inhibitors also induced the characteristic cytoplasmic and nuclear changes associated with apoptosis as assessed by transmission electron microscopy and DNA fragmentation as determined by flow cytometry. Taken together, these data show that apoptosis in HL-60 cells, induced by blocking arachidonate-phospholipid remodeling, is correlated with a redistribution of arachidonate in membrane phospholipids and suggest that such alterations represent a signal which controls the capacity of cells to proliferate. PMID- 8703925 TI - 5'-(p-fluorosulfonylbenzoyl)-2'(or 3')-(methylanthraniloyl)adenosine, fluorescent affinity labels for adenine nucleotide binding sites: interaction with the kinase active site of the receptor for epidermal growth factor. AB - We have found that the epidermal growth factor (EGF) receptor kinase can utilize the fluorescent ATP derivative, methylanthraniloyl ATP, as a substrate. On the basis of this observation, together with our previous studies that showed that 5' (p-fluorosulfonylbenzoyl)adenosine (5'-FSBAdo) is a highly specific affinity label for the ATP site of the kinase domain of the EGF receptor, we prepared new derivatives of 5'-FSBAdo, 5'-(p-fluorosulfonyl)-2'(or 3') (methylanthraniloyl)adenosine (FSBMantAdo), as fluorescent affinity labels for adenine nucleotide binding sites, and in particular for the ATP site of the EGF receptor. The two products were purified by HPLC and were characterized by UV-Vis absorbance spectroscopy, mass spectrometry, nuclear magnetic resonance spectroscopy, and fluorescence spectroscopy. Incubation of membrane vesicles containing the EGF receptor with either the 2' or 3' derivative resulted in irreversible inhibition of the receptor kinase activity, as assessed by autophosphorylation assays. Preincubation of vesicles with AMP imidodiphosphate (AMPPNP), a hydrolysis-resistant ATP analog, prior to treatment with FSBMantAdo resulted in the protection of the receptor kinase activity' from FSBMantAdo inactivation. Steady state fluorescence spectra (with excitation at 360 nm) revealed a blue shift in the emission maximum of partially purified FSBMantAdo labeled receptor (426 nm), as compared with the emission maximum of free FSBMantAdo (441 nm) in aqueous solution, suggesting that the receptor-bound label is in a relatively low polarity environment. These studies show that FSBMantAdo is a specific affinity label for the ATP site of the EGF receptor. FSBMantAdo may also prove useful as a fluorescent affinity label for other ATP binding sites. PMID- 8703926 TI - Phase partition analysis of nucleotide binding to axonemal dynein. AB - The binding of nucleoside triphosphates and related ligands to dynein ATPase from sea urchin sperm flagella has been studied by equilibrium partition analysis in an aqueous biphasic system containing dextran and poly(ethylene glycol). The stoichiometry of binding and the corresponding stepwise binding constants are obtained from direct binding isotherms fitted to the primary data. The results suggest that dynein possesses four different binding sites for nucleoside triphosphates per mole of heavy chain. The stepwise binding constants for MgATP range from approximately 10(4) M-1 to approximately 10(5) M-1. The isolated alpha and beta heavy chains have binding parameters similar to intact dynein. The amount of ADP bound normally is approximately 75% that of ATP, both for the intact dynein and for the separated heavy chains, although full saturation is achieved at high nucleotide concentrations. In the presence of the ATPase inhibitor vanadate, ADP binds with affinities similar to those of ATP, with binding constants close to those of ATP in the absence of vanadate. No appreciable binding of AMP or EDTA/ATP is observed. The substitution of Ca2+ or Fe3+ for Mg2+ does not significantly alter the amount of ATP bound; however, CaATP is bound with a somewhat lower affinity. Scatchard and Hill plots of the binding data and the calculated site-binding constants suggest that ATP and ADP bind in a weakly cooperative manner. These results suggest that the multiple binding of nucleotide to dynein heavy chains occurs at physiological concentrations, putatively at the four binding sites predicted earlier on the basis of their amino acid sequences. The data are consistent with a model in which, in addition to a single catalytic site, nucleotide binding occurs at additional noncatalytic sites that represent an as yet unknown functional aspect of dynein. PMID- 8703927 TI - Channel-forming activity of immunoaffinity-purified connexin32 in single phospholipid membranes. AB - Connexin32, a member of the family of proteins that forms gap junction channels between cells, was immunoaffinity-purified from rat liver using a monoclonal antibody, under nondenaturing conditions and reconstituted into unilamellar phospholipid liposomes and bilayers. Gel-filtration studies indicate that the connexin32 is purified predominantly in structures of a size consistent with that of single hemichannels and too small to be junctional channels (dimers of hemichannels). Purified connexin formed channels permeable to sucrose and to Lucifer Yellow. The permeability was reversibly reduced by acidic pH and unaffected by several agents that modulate coupling between cells. Modeling of the distribution of the permeability in the liposomes indicates that it is mediated by connexin structures that distribute among the liposomes as single hemichannels. Bilayer recordings of the purified connexin show high conductance channels with asymmetric voltage sensitivity. The results show that immunopurified connexin32 can form channels, in single phospholipid membranes, that have permeability similar to that of gap junction channels and thus can be utilized in studies of permeability and its regulation to investigate its role in normal physiological function, development, and disease. PMID- 8703928 TI - Evidence for a proximal histidine interaction in the structure of cytochromes c in solution: a resonance Raman study. AB - Soret-excited resonance Raman (RR) spectra of oxidized and reduced cytochromes c from Rhodospirillum molischianum and Rhodobacter sphaeroides, in solution, are reported. The spectra of the type I ferricytochromes c in both species contain different extents of two forms. One of these is readily assignable to a "normal" five-coordinated high-spin heme. The second species with v3 and v10 modes at 1502 and 1635 cm-1, respectively, is attributed to a five-coordinated intermediate spin heme. The RR data show that the equilibrium between these two forms is species-dependent at neutral pH and 20 degrees C. The v(Fe-His) mode of the a form of reduced cytochromes c is assigned to a band at 228-231 cm-1, indicating that the proximal His has a strong electronegative character. X-ray crystallographic data on R. molischianum ferricyt c show that the proximal His has no interaction with either the protein or water molecules [Finzel, B.C., Weber, P.C., Hardman, K.D., & Salemme, F.R.(1985) J. Mol. Biol. 186, 627-643]. Considering that the absence of H bonding at the coordinated histidine corresponds to a low frequency for the v(Fe-His) mode (195-205 cm-1), the structure and/or environment of the proximal histidine appears different for cyt c (III) in the crystal and cyt c (II) in aqueous solution. To account for the elevated frequency of the v(Fe-His) mode of cyt c (II), several possibilities have been examined. Among these, we propose that a conserved Lys residue, located in the protein sequence three residues before the His ligand, can form an electrostatic interaction with the (His)N1 atom, directly or through a water molecule. It is further suggested that this electrostatic interaction could also play a role in the high-spin <--> intermediate-spin equilibrium of oxidized cytochromes c. PMID- 8703929 TI - Electron transfer dynamics of Rhodopseudomonas viridis reaction centers with a modified binding site for the accessory bacteriochlorophyll. AB - Femtosecond spectroscopy in combination with site-directed mutagenesis was used to study the influence of histidine L153 in primary electron transfer in the reaction center of Rhodopseudomonas viridis. Histidine was replaced by cysteine, glutamate, or leucine. The exchange to cysteine did not lead to significant changes in the primary reaction dynamics. In the case of the glutamate mutation, the decay of the excited electronic level of the special pair P* is slowed by a factor of 3. The exchange to leucine caused the incorporation of a bacteriopheophytin b instead of a bacteriochlorophyll b molecule at the BA site. As a consequence of this chromophore exchange, the energy level of the electron transfer state P+BA- is lowered to such an extent that repopulation from the next electron transfer intermediate state P+HA- takes place, resulting in a long lasting P+BA- population. The observed differences in time constants are discussed in the scope of nonadiabatic electron transfer theory considering the influence of the amino acids at position L153 and the chromophore exchange on the energy level of the intermediate state P+BA-. The results show that the high efficiency of primary electron transfer is reduced substantially, if the energy level of P+BA- is lowered or raised by several hundred wave numbers. PMID- 8703930 TI - Control of the integral membrane proton pump, bacteriorhodopsin, by purple membrane lipids of Halobacterium halobium. AB - Brief exposure of purple membrane (PM) to dilute Triton X-100 eliminates the actinic light effect on the relative amounts of fast M (Mf) and slow M (Ms) intermediates and alters the character and kinetics of the photocycle, without destroying the native BR trimers (Mukhopadhyay et al., 1994). Particular membrane lipids are removed during the Triton treatment, and adding back an extract of membrane lipids can repair most of the affected photocycle behavior (Dracheva et al., 1996). This paper defines conditions which are important in the reconstitution procedure, using a group of quantitative parameters which measure the extents of damage and repair. Circular dichroism in both the UV and visible ranges shows that Triton can disturb both the secondary structure of BR and its ability to polymerize into trimers. Whereas the damage to protein conformation could be reversed by lipids alone, the formation of trimers and recovery of normal photocycle behavior required both lipids and a high salt concentration. PMID- 8703931 TI - Role of adenosine 5'-triphosphate hydrolysis in the assembly of the bacteriophage T4 DNA replication holoenzyme complex. AB - Steady-state and pre-steady-state rates of ATP hydrolysis by the 44/62 accessory protein were determined to elucidate the role of ATP hydrolysis in bacteriophage T4 holoenzyme complex formation. Steady-state ATPase measurements of the 44/62 protein under various combinations of 45 protein, DNA substrate, and T4 exo- polymerase indicate that although the 44/62 protein synergistically hydrolyzes ATP in the presence of 45 protein and DNA substrate, the ATPase activity of 44/62 is diminished substantially upon the formation of the holoenzyme complex. The decrease in activity is primarily in kcat while the K(m) for ATP is changed unsubstantially by the various combinations. Data suggest that the decrease in the rate of ATP hydrolysis upon the addition of T4 exo- polymerase in the presence of 45 protein and DNA substrate is due to formation of a stable holoenzyme complex consisting of only the 45 protein and T4 exo- polymerase in a 1:1 ratio. The 44/62 protein acts catalytically to load 45 protein onto the DNA substrate and does not remain a component of the holoenzyme complex. Pre-steady state kinetic analysis of the ATP hydrolysis reaction catalyzed by the 44/62 protein loading the 45 protein onto the DNA substrate in the absence or presence of polymerase is biphasic, in which a burst in ATP hydrolysis precedes the steady state rate of ATP hydrolysis. An identical burst in ATP consumption is obtained under either condition, indicating that ATP hydrolysis is not required to load polymerase into the holoenzyme complex. The data suggest one turnover of ATP at each of the four ATPase active sites of the 44/62 protein per 45 protein loaded. ATP hydrolysis by the 44/62 protein under conditions of holoenzyme complex formation is the rate-limiting step in holoenzyme complex formation. The process of holoenzyme formation appears to be identical for leading and lagging strand synthesis. PMID- 8703932 TI - Role of calf RTH-1 nuclease in removal of 5'-ribonucleotides during Okazaki fragment processing. AB - The role of the exonucleolytic activity of the calf 5' to 3' exo/endonuclease, a RAD2 homolog 1 (RTH-1) class nuclease, in lagging-strand DNA replication has been examined using model Okazaki fragment substrates. These substrates exemplify the situation in Okazaki fragment processing which occurs after the initiator RNA primer is cleaved off, and released intact, by calf RNase HI, leaving a single ribonucleotide at the 5' end of the RNA-DNA junction. This final RNA is then removed by the calf RTH-1 nuclease [Turchi et al. (1994) Proc. Natl. Acad. Sci. U.S.A. 91, 9803-9807]. The cleavage specificity of calf RTH-1 nuclease for different junction ribonucleotides was compared. These were removed without the usual requirement of calf RTH-1 for an immediately adjacent upstream primer. In most cases, the presence of an upstream DNA or RNA primer, separated from the monoribonucleotide-DNA segment by either a nick or a gap, reduced the efficiency of removal of the monoribonucleotide compared to the removal seen with no upstream primer. Substrates in which the monoribonucleotide-DNA segment had been replaced by an oligomer of the same sequence but consisting entirely of DNA also exhibited upstream primer inhibition. Results with various sequences indicated that the upstream primer is generally inhibitory for ribonucleotide removal but is sometimes neutral. For deoxynucleotide removal it could be stimulatory, neutral, or inhibitory. Possible reasons for the unexpected lack of upstream primer dependence have been explored. The ratio of RNase HI to RTH-1 was also shown to be critical for both enzymes to work together efficiently. These results suggest that regions of upstream primer inhibition within the genome may play a role in determining the mechanism by which mammalian Okazaki fragments are processed. PMID- 8703934 TI - Evidence for an essential lysyl residue in phospholipase D from Streptomyces sp. by modification with diethyl pyrocarbonate and pyridoxal 5-phosphate. AB - Diethyl pyrocarbonate inactivated phospholipase D from Streptomyces PMF with second-order rate constants of 0.7 M-1 s-1 at pH 6.1 or 222 M-1 s-1 at pH 8.3 and 25 degrees C, and modified 5 His residues per enzyme molecule. The His residues, however, were not essential for activity because: (a) the second-order rate constants for reaction of diethyl pyrocarbonate with the His residues of the enzyme, which were 1.4 M-1 s-1 at pH 6.1 or 7.2 M-1 s-1 at pH 8.3 and 25 degrees C, differed, both at low and high pH values, from the inactivation rates, and (b) the reversal of His modification by hydroxylamine was not accompanied by recovery of activity. As demonstrated by dinitrophenylation experiments carried out on the treated enzyme, diethyl pyrocarbonate also modified up to 20 Lys residues per enzyme molecule. Other amino acid residues and the conformation and hydrodynamic volume of the enzyme were not modified. The involvement of a Lys residue in enzyme activity was confirmed through experiments with pyridoxal 5-phosphate which inactivated phospholipase D, after NaBH4 reduction, with a second-order rate constant of 3.5 M-1 s-1 at pH 8.5 and 15 degrees C. The inactivation took place with concomitant modification of 4 Lys residues, only one of which was found to be essential using the kinetic method of Tsou (Tsou, C.-L. (1962) Sci. Sin. 11, 1535-1538). Dicaproyl phosphatidylcholine markedly protected the enzyme against inactivation by DEP or PLP, and this strongly suggests that the essential Lys residue is located in or near the substrate binding site. PMID- 8703933 TI - Evidence for flavin movement in the function of p-hydroxybenzoate hydroxylase from studies of the mutant Arg220Lys. AB - The isoalloxazine ring system of the FAD cofactor of p-hydroxybenzoate hydroxylase must be secluded from solvent at specific stages of catalysis in order to form and stabilize a flavin C4a-hydroperoxide. This species may then react with the activated phenolate of p-hydroxybenzoate. A number of crystal structures of the enzyme with alterations to active site substituents or complexes with analogue benzoates have revealed an alternate position for the isoalloxazine (Gatti et al. (1994) Science 266, 110-114; Schreuder et al. (1994) Biochemistry 33, 10161-10170). This new flavin conformation is 7 A "out" toward solvent and may open a passage for substrate entry to the active site. Arginine 220 is one of the few residues in the structure to demonstrate conformational changes when the flavin is "out". In this study we have made the Arg220Lys mutant to test the significance of this residue in flavin movement. The R220K mutation has brought about dramatic alterations to all aspects of catalysis. Stopped flow kinetic characterization of the mutant has revealed that, while the effector role for the substrate is maintained, there exists an order of magnitude decrease in the limiting rate of reduction, even though there is 40-fold increase in association with NADPH. The mutant enzyme has only a fraction of its reductive half-reaction coupled to product formation, and the hydroxylation process is slow. This occurs despite a higher proportion of the more activated substrate phenolate in the active site. Many of the observed changes can be attributed to a decrease in the stability of the "in" conformation of the flavin during the catalysis and indicate a role for flavin conformational states in many of the catalytic processes of the enzyme. PMID- 8703935 TI - NMR analysis of site-specific ligand binding in oligomeric proteins. Dynamic studies on the interaction of riboflavin synthase with trifluoromethyl substituted intermediates. AB - The binding of small ligands to symmetrical oligomeric proteins may lead to a number of different partially ligated intermediates but should finally yield a symmetrical fully ligated enzyme/ligand complex. In the case of the trimeric protein, riboflavin synthase, some ligands form an unexpected protein/ligand complex, even in the presence of a large excess of ligand. Three different bound forms were observed by 19F NMR spectroscopy, and Scatchard-type analysis suggested binding sites of similar affinities. NOESY analysis of the kinetic network revealed that the three bound states exchange with free ligand, but not with each other, thus suggesting that the trimeric enzyme could be asymmetrical. This information permits appropriate precautions to be taken during X-ray structure analysis of riboflavin synthase, which is in progress. Quantitative analysis of the NOESY spectra yielded different rate constants for the different binding sites. For comparison, the monomeric lumazine protein was investigated as an example of a case with simple two-site exchange. For such systems, all kinetic parameters including kon and the dissociation constant can be determined from the NOESY spectrum. The data show that NMR spectroscopy can produce qualitative and quantitative information in cases of nonequivalent binding sites in oligomeric proteins if isolated NMR signals of the different forms can be observed. The technique is not limited to 19F as reporter nucleus. PMID- 8703936 TI - Trimeric assembly and three-dimensional structure model of the FACIT collagen COL1-NC1 junction from CD and NMR analysis. AB - The 3D structure of the COL1-NC1 junction of FACIT type XIV collagen was investigated using GYCDPSSCAG and (GPP*)3GYCDPSSCAG synthetic peptides, circular dichroism, and NMR. At -20 degrees C and under air oxidation catalyzed by Cu2+, the peptide (GPP*)3GYCDPSSCAG is able to self-associated with high yield into a stable triple disulfide bonded trimer. The presence of a triple helical conformation was confirmed by circular dichroism. The analysis of the trimer by 2D NMR provided a set of distance constraints for the noncollagenous part. Molecular models for the 3D structure of COL1-NC1 junction were calculated, using the NMR distance constraints in combination with the 3D structural data recently established by X-ray crystallography [Bella, J., Eaton, M., Brodsky, B., & Berman, H. M. (1994) Science 266, 75-81] for a collagenous triple helix. From the eight theoretically possible arrangements for the three interchain disulfide bonds, only two close disulfide conformers are compatible with the experimental data. The main feature of the trimer structure is the asymmetry of the molecule due to the disulfide bond pattern that induces a particular folding of one chain. This chain forms a turn-like structure locked by two disulfide bonds with the two other chains. The turn-like folding is close to that observed for the cyclized oxidized monomeric peptide. This is the first report of the 3D structure model for a junction between a collagenous triple helical domain and a noncollagenous domain. PMID- 8703937 TI - Backbone dynamics of the C-terminal domain of Escherichia coli topoisomerase I in the absence and presence of single-stranded DNA. AB - The backbone dynamics of the C-terminal DNA-binding domain of Escherichia coli topoisomerase I has been characterized in the absence and presence of single stranded DNA by NMR spectroscopy. 15N spin-lattice relaxation times (T1), spin spin relaxation times (T2), and heteronuclear NOEs were determined for the uniformly 15N-labeled protein. These data were analyzed by using the model-free formalism to derive the model-free parameters (S2, tau e, and R(ex)) for each backbone N-H bond vector and the overall molecular rotational correlation time (tau m)., The molecular rotational correlation time tau m was determined to be 7.49 +/- 0.36 ns for the free and 12.7 +/- 1.07 ns for the complexed protein. Several residues were found to be much more mobile than the average, including 11 residues at the N-terminus, 2 residues at the C-terminus, and residues 25 and 31 35 which are located in a region of the protein that binds to DNA. The binding of ssDNA to the free protein causes a slight increase in the order parameters (S2) for a small number of residues and a slight decrease in the order parameters (S2) for the majority of the residues. In particular, upon binding to ssDNA, the mobility of the first alpha-helix and the two beta-sheets was slightly increased, and the mobility of a few specific residues in the loops/turns was restricted. These results differ from the previous studies on the backbone dynamics of molecular complexes in which reduced mobilities were typically observed upon ligand binding. PMID- 8703938 TI - A mutational analysis of the binding of two different proteins to the same antibody. AB - The crystal structures of the complexes between the anti-hen egg white lysozyme (HEL) antibody D1.3 and HEL and between D1.3 and the anti-D1.3 antibody E5.2 have shown that D1.3 contacts these two proteins through essentially the same set of combining site residues [Fields, B. A., Goldbaum, F. A., Ysern, X., Poljak, R. J., & Mariuzza, R. A. (1995) Nature 374, 739-742]. To probe the relative contribution of individual residues to complex stabilization, single alanine substitutions were introduced in the combining site of D1.3, and their effects on affinity for HEL and for E5.2 were measured using surface plasmon resonance detection, fluorescence quench titration, or sedimentation equilibrium. The energetics of the binding to HEL are dominated by only 3 of the 13 contact residues tested (delta Gmutant-delta Gwild type > 2.5 kcal/mol): VLW92, VHD100, and VHY101. These form a patch at the center of the interface and are surrounded by residues whose apparent contributions are much less pronounced ( < 1.5 kcal/mol). This contrasts with the interaction of D1.3 with E5.2 in which most the contact residues (11 of 15) were found to play a significant role in ligand binding ( > 1.5 kcal/mol). Furthermore, even though D1.3 contacts HEL and E5.2 in very similar ways, the functionally important residues of D1.3 are different for the two interactions, with only substitutions at D1.3 positions VH100 and VH101 greatly affecting binding to both ligands. Thus, the same protein may recognize different ligands in ways that are structurally similar yet energetically distinct. PMID- 8703939 TI - Solution structure of (rGCGGACGC)2 by two-dimensional NMR and the iterative relaxation matrix approach. AB - The three-dimensional solution structure of the RNA self-complementary duplex [sequence: see text] was derived from two-dimensional NMR and the iterative relaxation matrix approach. Each GA mismatch forms two hydrogen bonds: A-NH6 to G O6 and A-N1 to G-NH1 (imino). This is the first three-dimensional RNA structure with imino hydrogen-bonded tandem GA mismatches. This GA structure is totally different from the sheared tandem GA structure in [sequence: see text] which also has two hydrogen bonds: A-N7 to G-NH2 and A-NH6 to G-N3 [SantaLucia, J., Jr., & Turner, D. H. (1993) Biochemistry 32, 12612-12623]. In particular, the sheared and imino GA mismatches produce a narrowing and widening of the backbone, respectively. The results show that substitutions of Watson-Crick base pairs can have dramatic effects on the three-dimensional structures of adjacent non-Watson Crick paired regions; i.e., the structure depends on sequence context. Thus compensating substitutions in site-directed mutagenesis experiments may not always restore biological activities. PMID- 8703940 TI - Structure-based understanding of ligand affinity using human thrombin as a model system. AB - Kinetic study of a series of compounds containing the thrombin-directed peptide D Phe-ProboroArg-OH had indicated that the structure of the N-terminal blocking group may be correlated with binding [Kettner, C., Mersinger, L., & Knabb, R. (1990) J. Biol. Chem. 265, 18289-18297]. In order to further study this phenomenon, a second series of compounds that contains a C-terminal methyl ester in place of the boronic acid was synthesized, binding measured, and the three dimensional structure in complex with human thrombin determined by X-ray crystallography. Incubation of Ac-D-Phe-Pro-Arg-OMe, Boc-D-Phe-Pro-Arg-OMe, and H D-Phe-Pro-Arg-OMe resulted in the formation of thrombin-product complexes within the crystal. Ki values for the corresponding products (free carboxylic acids) were 60 +/- 12 microM, 7.8 +/- 0.1 microM, 0.58 +/- 0.02 microM, respectively, indicating that the nature of the N-terminal blocking group has a significant effect on affinity. Examination of the crystal structures indicated that the higher affinity of the H-D-Phe peptide is due to rearrangement of one residue comprising the S3 site (Glu192) in order to maximize electrostatic interactions with the "NH3(+)-" of H-D-Phe. The relative affinity of Boc-D-Phe-Pro-Arg-OH is due to favorable hydrophobic interactions between thrombin and the bulky butyl group. However, this results in less favorable binding of Arg-P1 in the oxyanion hole as shown by long hydrogen-bonding distances. This work gave rise to some general observations applicable to structure-based drug design: (1) altering the structure of an inhibitor at one site can affect binding at an unchanged distal site; (2) minor alteration of inhibitor structure can lead to small, but significant reorganization of neighboring protein structure; (3) these unexpected reorganizations can define alternate binding motifs. PMID- 8703941 TI - The plasma and cytoplasmic forms of human gelsolin differ in disulfide structure. AB - Gelsolin is a widely distributed actin binding protein that regulates actin filament length. It exists in both an intracellular and an extracellular form that is derived from a single gene by alternative splicing. Both forms contain the six homologous domains that are responsible for function. Little is known regarding differences between the forms. We have used a combination of cysteine specific modification with 4-vinylpyridine, HPLC peptide mapping methods, and mass spectrometry to analyze the disulfide structures of human plasma and cytoplasmic gelsolin. Of the five Cys residues in the human gelsolin sequence, all were present in the free thiol form in human cytoplasmic gelsolin, while only three of them were free thiols in the human plasma form. Cys residues 188 and 201 in domain 2 of plasma gelsolin were disulfide linked. Recombinant human plasma gelsolin that had been expressed intracellularly in Escherichia coli and as a secreted protein from Cos green monkey cells was also investigated. The E. coli product lacked the disulfide but could be converted to the plasma-like structure with mild oxidation while the mammalian product formed the correct disulfide prior to isolation. Structural differences were also detected by limited proteolysis with plasmin. The differences in proteolytic susceptibility were also due to perturbations in domain 2. These studies demonstrate that the intracellular and extracellular gelsolins are structurally distinct and suggest that at least some of the preparations of recombinant gelsolin that are being used to study structure/function may be improperly folded. The experiments also demonstrate a general method for the location of disulfide bonds in proteins. PMID- 8703942 TI - Functional site in alpha-lactalbumin encompasses a region corresponding to a subsite in lysozyme and parts of two adjacent flexible substructures. AB - Aromatic cluster 1 of alpha-lactalbumin (LA), a substructure adjacent to the cleft, is important for its interaction with galactosyltransferase (GT) and effects on glucose binding in the lactose synthase complex [Grobler, J. A., Wang, M., Pike, A. K., & Brew, K. (1994) J. Biol. Chem. 269, 5106-5114]. The full extent of the functional region in LA has been probed by mutagenesis of residues that are near aromatic cluster 1 or within the cleft that corresponds to the active site in the homologous type c lysozymes. The conserved residues Val42, Gln54, and Ile59, which correspond to residues of lysozyme that act in substrate binding in subsites C to E, together with residues adjacent to aromatic cluster 1, were found to be not required for activity. In contrast, replacing Leu110, a component of the region corresponding to lysozyme subsite F, with His or Glu greatly reduces the affinity of LA for GT while the introduction of Arg lowers the synergism of LA and glucose binding to GT and also reduces the affinity of LA for GT. Substitutions for Ala106, which is adjacent to Leu110 in the structure, also perturb activity. The region of the cleft corresponding to subsite F is important for function in LA as well as in lysozyme since other components of this subsite, His32 and Phe31, are also crucial for LA activity. The qualitatively different effects of various substitutions for Leu110 may be mediated by their influence on His32 or by changes in the structure of the lactose synthase complex. PMID- 8703943 TI - Crystal structure of the complex of UMP/CMP kinase from Dictyostelium discoideum and the bisubstrate inhibitor P1-(5'-adenosyl) P5-(5'-uridyl) pentaphosphate (UP5A) and Mg2+ at 2.2 A: implications for water-mediated specificity. AB - The three-dimensional structure of the UMP/CMP kinase (UK) from the slime mold Dictyostelium discoideum complexed with the specific and asymmetric bisubstrate inhibitor P1-(5'-adenosyl) P5-(5'-uridyl) pentaphosphate (UP5A) has been determined at a resolution of 2.2 A. The structure of the enzyme, which has up to 41% sequence homology with known adenylate kinases (AK), represents a closed conformation with the flexible monophosphate binding domain (NMP site) being closed over the uridyl moiety of the dinucleotide. Two water molecules were found within hydrogen-bonding distance to the uracil base. The key residue for the positioning and stabilization of those water molecules appears to be asparagine 97, a residue that is highly specific for AK-homologous UMP kinases, but is almost invariably a glutamine in adenylate kinases. Other residues in this region are highly conserved among AK-related NMP kinases. The catalytic Mg2+ ion is coordinated with octahedral geometry to four water molecules and two oxygens of the phosphate chain of UP5A but has no direct interactions with the protein. The comparison of the geometry of the UKdicty.UP5A.Mg2+ complex with the previously reported structure of the UKyeast.ADP.ADP complex [Muller-Dieckmann & Schulz (1994) J. Mol. Biol. 236, 361-367] suggests that UP5A in our structure mimics an ADP.Mg.UDP biproduct inhibitor rather than an ATP. MG.UMP bisubstrate inhibitor. PMID- 8703944 TI - Topological folding and proteolysis profile of P-glycoprotein in membranes of multidrug-resistant cells: implications for the drug-transport mechanism. AB - P-glycoprotein (Pgp)1 is a polytopic membrane protein and functions as an energy dependent drug efflux pump. It is responsible for multidrug resistance (MDR) in cancer cell lines. Recently, the topological structure of Pgp has been investigated. However, the results are in dispute. A major question concerning the Pgp topology is the membrane orientation of the loop linking TM4 and TM5 (loop 4) and the loop linking TM8 and TM9 (loop 8). In this study, we generated polyclonal antibodies specific to these two loops. In combination with a panel of other well-characterized site-specific polyclonal- and monoclonal antibodies of Pgp, we tested the membrane orientation of these two loops of Pgp in multidrug resistant cells using immunocytochemistry and proteolysis/membrane protection assay. Our results showed that (1) both loops 4 and 8 are located extracellularly whereas other domains, such as the ATP-binding sites, are in the cytoplasm and (2) proteolysis of Pgp is not a random event and the trypsin-sensitive sites are cleaved in orders. Since the Pgp was not genetically manipulated in this study, in contrast to previous studies, we believe that naturally expressed Pgp molecules have an unconventional topology. We speculate that this alternate topology of Pgp may represent a different functional state of the protein. Further detailed analysis of Pgp topology will help to understand the fundamental mechanism of drug transport by Pgp. PMID- 8703946 TI - Coordinate regulation of glucocorticoid receptor and c-jun gene expression is cell type-specific and exhibits differential hormonal sensitivity for down- and up-regulation. AB - We have previously proposed a novel mechanism for the coupled regulation of glucocorticoid receptor (GR) and c-jun transcription in triamcinolone acetonide (TA)-treated AtT-20 cells. This involved transcriptional interference of AP-1 (Fos/Jun)-driven gene transcription by the formation of inactive GR/Jun heterodimers. To further elucidate the molecular mechanism for GR autoregulation, the expression of GR and c-jun mRNA and protein levels were examined in both mouse L929 fibroblast cells and human CEM-C7 acute lymphoblastic leukemia cells. A rapid down-regulation of both GR and c-jun mRNA and protein levels occurs in TA treated L929 cells. All-trans-retinoic acid (RA) treatment of Jun-deficient, mouse F9, teratocarcinoma cells causes the induction of c-jun expression. The increased expression of both c-jun mRNA and protein is accompanied by the induction of GR expression. These data further suggest that functional cJun is needed for the expression of the GR and c-jun genes in F9 cells. CEM-C7 cells undergo apoptosis after exposure to glucocorticoids. There is a parallel up regulation of GR and c-jun mRNA levels in TA-treated CEM-C7 cells. This is accompanied by a concomitant increase in GR and cJun protein levels. Dose response analyses reveal the expected coordinate regulation of both GR and c-jun mRNA and protein in L929 cells (decreasing) and in CEM-C7 cells (increasing). However, approximately 20-fold less TA is required for the inhibition of GR and c jun expression as compared to that required for the stimulation of these two genes. These data demonstrate that the coordinate regulation of GR and c-jun gene expression is dose-dependent and cell type-specific. These results, along with previously reported data, suggest that GR complex formation with itself or with another transcription factor is important for the coordinate up- and down regulation, respectively, of the GR and c-jun genes. PMID- 8703945 TI - A drug resistance mutation in the inhibitor binding pocket of human immunodeficiency virus type 1 reverse transcriptase impairs DNA synthesis and RNA degradation. AB - Selection of the IIIB strain of human immunodeficiency virus type (HIV-1) resistant to the (alkylamino)piperidine-bis(heteroaryl)piperazine (AAP-BHAP) U 104489 results in substitution of a glycine to glutamate at residue 190 (G190E) of reverse transcriptase (RT). The AAP-BHAP resistant HIV-1 displays reduced in vitro replication capacity [Olmsted, R. A., et. al. (1966) J. Virol. 70, 3698 3705]. We report here that the G190E mutation in recombinant heterodimeric HIV-1 RT, compared to the wild-type RT (G190) or a G190A control mutant, results in a 40% and 80% reduction in the polymerase and RNase H specific enzymatic activities, respectively. A primer-extension assay that allowed determination of DNA elongation by the G190E mutant RT on a heteropolymeric HIV-1 gag-based RNA template showed an overall decrease in DNA polymerization. The size distribution of products generated by G190E RT-associated RNase H digestion of RNA from [35S]poly(rA).poly(dT) was markedly distinct from that of the G190A RT and was consistent with the observed reduction in RT-associated RNase H activity of the G190E RT. When challenged with unlabeled substrates, the G190E RT was relatively nonprocessive with respect to DNA synthesis and RNA degradation. It is concluded that the deleterious effect of the G190E resistance mutation on both of these RT functions is most likely involved in the observed retarded replication capacity of the AAP-BHAP-(U-104489-) resistant HIV-1. PMID- 8703947 TI - Chemical synthesis of a Zwitterhormon, insulaxin, and of a relaxin-like bombyxin derivative. AB - The structural motif of insulin and relaxin is frequently seen in molecules of divergent functions and origins. The insect developmental factor bombyxin, the relaxin-like factor from Leydig cells, and the insulin-like factor 4 (INSL4) all are made of two disulfide-linked chains and have one disulfide bond within the A chain. The polyclonal antibody R6, which was raised against porcine relaxin, reacts with a wide variety of naturally occurring relaxins from primates, marine and terrestrial mammals, and elasmobranchs but does not recognize insulin. The antibody binds mainly to the arginines that occur in the N, N+4 positions in the B-chains of all relaxins which also constitute the receptor-binding site. The receptor-binding haptens were incorporated by total synthesis into human despentapeptide insulin and bombyxin II, a developmental factor from the silk moth Bombyx mori. In the process the insect factor became a perfect antigen for the anti-relaxin antibody, whereas the human insulin was transformed into a bona fide relaxin. The conversion was affected by changing four critical residues so that the insulin activity was retained to the extent of 10% of the original level. This, to the best of our knowledge, is the first designer protein to incorporate two unrelated biological functions in one primary sequence, and we are therefore proposing that, analogous to zwitterion, the generic name "Zwitterhormon" (German spelling) be used for this type of construct. PMID- 8703948 TI - Catalysis of disulfide isomerization in thrombospondin 1 by protein disulfide isomerase. AB - Thrombospondin 1 is a multidomain glycoprotein from platelets and most cells that participates in diverse biological processes. The structure and some functional properties of thrombospondin 1 are regulated by disulfide interchange in the Ca(2+)-binding repeats and C-globular domain. The recent identification of the enzyme, protein disulfide isomerase, on the platelet surface suggested that protein disulfide isomerase may catalyze disulfide isomerization in platelet thrombospondin 1. Protein disulfide isomerase was found to form disulfide-linked complexes with thrombospondin 1, which is consistent with protein disulfide isomerase-mediated rearrangement of disulfide bonds in thrombospondin 1. To quantitate disulfide interchange in thrombospondin 1, perturbation of the enzyme inhibitory properties of platelet thrombospondin 1 were measured, specifically changes in the apparent dissociation constant for inhibition of neutrophil cathepsin G by thrombospondin 1. The inhibition constant increased > or = 10-14 fold following incubation of either Ca(2+)-replete or Ca(2+)-depleted thrombospondin 1 with protein disulfide isomerase and reduced glutathione. The rate of protein disulfide isomerase-catalyzed disulfide interchange in thrombospondin 1 increased linearly with protein disulfide isomerase concentration and the K(m) for reduced glutathione was 0.4 +/- 0.2 mM. Disulfide isomerization in both platelet and fibroblast thrombospondin 1 was probed by measuring perturbation in epitopes for two anti-thrombospondin 1 monoclonal antibodies. Antibody D4.6 binds to the C-terminal Ca(2+)-binding domains which are involved in disulfide interchange, whereas antibody HB8432 binds toward the N terminus of the thrombospondin 1 subunit. In accordance with the location of these epitopes, incubation of platelet thrombospondin 1 or fibroblast thrombospondin 1 with protein disulfide isomerase and reduced glutathione resulted in 2-fold enhancement of binding of D4.6, whereas binding of HB8432 did not significantly change. In summary, protein disulfide isomerase catalyzes disulfide interchange in thrombospondin 1 which alters binding of neutrophil cathepsin G and antibody D4.6 to thrombospondin 1. PMID- 8703949 TI - Enzymological characterization of the Pasteurella multocida hyaluronic acid synthase. AB - Hyaluronic acid (HA), a linear polysaccharide composed of alternating glucuronic acid and N-acetylglucosamine residues, is an essential molecule of higher vertebrates. The fowl cholera pathogen Pasteurella multocida Carter Type A also produces HA in the form of an extracellular capsule in order to evade host defenses. HA synthase activity could be obtained from cell-free membrane preparations of P. multocida. The enzyme utilized UDP-sugar precursors of HA in the presence of Mg2+ or Mn2+ at neutral pH. Mn2+ at 1 mM stimulated approximately 2-fold more incorporation than Mg2+ at 10 mM. On the other hand, the analogous enzyme from group A Streptococcus, HasA, is stimulated more by Mg2+ than Mn2+. The apparent Michaelis constants, K(M), of the P. multocida HA synthase for UDP-N acetylglucosamine and UDP-glucuronic acid were estimated to be approximately 75 and approximately 20 microM, respectively, in the presence of Mg2+, which suggests that the substrates are bound with 2-3-fold higher affinity than by the HasA enzyme. The rate enhancement observed with Mn2+ is apparently not due to better binding of the sugar nucleotide precursors complexed to Mn ion because the K(M) value, a measure of substrate affinity, increases by 25-50% in comparison to Mg2+. In summary, the HA synthase from P. multocida, a Gram-negative bacterium, has kinetic optima distinct from those of HasA, the analog from the Gram-positive group A Streptococcus. PMID- 8703950 TI - Kinetic mechanisms of polyphosphate glucokinase from Mycobacterium tuberculosis. AB - Polyphosphate glucokinase from Mycobacterium tuberculosis catalyzes the phosphorylation of glucose using inorganic polyphosphates [poly(P)] or ATP. The steady-state kinetic mechanisms of the poly(P)- and ATP-dependent glucokinase reactions were investigated using initial velocity, product inhibition, and dead end inhibition analyses. In the poly(P)-dependent reaction, the enzyme follows an Ordered Bi Bi sequential mechanism with poly(P) binding to the enzyme first and glucose 6-phosphate dissociating last. Polyphosphate is utilized nonprocessively with a preference for longer chains due to higher kcat/K(m) values. The lack of inhibition at high poly(P) concentrations suggests that binding of poly(P) as a product is not favorable. In the ATP-dependent glucokinase reaction, the data are also consistent with an Ordered Bi Bi sequential mechanism, with ATP binding to the enzyme first and glucose 6-phosphate leaving last. At high concentrations, ATP displays competitive substrate inhibition with respect to glucose, which is consistent with the formation of an enzyme.ATP.ATP nonproductive complex. The overall catalytic efficiencies (kcat/KiaK(b)) of the poly(P)- and ATP-dependent reactions are approximately 10(11) M-2 s-1 and approximately 10(8) M-2 s-1, respectively. The higher catalytic efficiency, high value of the substrate specificity constant (kcat/K(a)) approaching a diffusion-controlled limit, and the absence of substrate inhibition in the poly(P)-dependent reaction suggest that poly(P), rather than ATP, is the major phosphate donor for poly(P) glucokinase in M. tuberculosis. PMID- 8703951 TI - Mechanism of aldehyde oxidation catalyzed by horse liver alcohol dehydrogenase. AB - The mechanism of oxidation of benzaldehyde to benzoic acid catalyzed by horse liver alcohol dehydrogenase (HLADH) has been investigated using the HLADH structure at 2.1 A resolution with NAD+ and pentafluorobenzyl alcohol in the active site [Ramaswamy et al. (1994) Biochemistry 33,5230-5237]. Constructs for molecular dynamics (MD) investigations with HLADH were obtained by a best-fit superimposition of benzaldehyde or its hydrate on the pentafluorobenzyl alcohol bound to the active site Zn(II)ion. Equilibrium bond lengths, angles, and dihedral parameters for Zn(II) bonding residues His67, Cys46, and Cys174 were obtained from small-molecule X-ray crystal structures and an ab initio-derived parameterization of zinc in HLADH [Ryde, U. (1995) Proteins: Struct., Funct., Genet. 21,40-56]. Dynamic simulations in CHARMM were carried out on the following three constructs to 100 ps: (MD1) enzyme with NAD+, benzaldehyde, and zinc ligated HO-in the active site; (MD2) enzyme with NAD+ and hydrated benzaldehyde monoanion bound to zinc via the pro-R oxygen, with a proton residing on the pro-S oxygen; and (MD3) enzyme with NAD+ and hydrated benzaldehyde monoanion bound to zinc via the pro-S oxygen, with a proton residing on the pro-R oxygen. Analyses were done of 800 sample conformations taken in the last 40 ps of dynamics. Structures from MD1 and MD3 were used to define the initial spatial arrangements of reactive functionalities for semiempirical PM3 calculations. Using PM3, model systems were calculated of ground states and some transition states for aldehyde hydration, hydride transfer, and subsequent proton shuttling. With benzaldehyde and zinc-bound hydroxide ion in the active site, the oxygen of Zn(II)-OH resided at a distance of 2.8-5.5 A from the aldehyde carbonyl carbon during the dynamics simulation. This may be compared to the PM3 transition state for attack of the Zn(II)-OH oxygen on the benzaldehyde carbonyl carbon, which has an O...C distance of 1.877 A. HLADH catalysis of the aldehyde hydration would require very little motion aside from that in the ground state. Two simulations of benzaldehyde hydrate ligated to zinc (MD2 and MD3) both showed close approach of the aldehyde hydrate hydrogen to NAD+C4, varying from 2.3 to 3.3 A, seemingly favorable for the hydride transfer reaction. The MD2 configuration does not allow proton shuttling. On the other hand, when the pro-S oxygen is ligated to zinc (MD3), the proton on the pro-R oxygen averages 2.09 A from the hydroxyl oxygen of Ser48 such that initiation of shuttling of protons via Ser48 to the ribose 2'-hydroxyl oxygen to the 3'-hydroxyl oxygen to His51 nitrogen is sterically favorable. PM3 calculations suggest that this proton shuttle represents a stepwise reaction which occurs subsequent to hydride transfer. The PM3 transition state for hydride transfer based on the MD3 configuration has the transferring hydride 1.476 A from C4 of NAD+ and 1.433 A from the aldehyde alpha-carbon. PMID- 8703952 TI - Stereospecificity of thermostable ornithine 5-aminotransferase for the hydrogen transfer in the L- and D-ornithine transamination. AB - The thermostable ornithine 5-aminotransferase of a thermophile, Bacillus sp. YM 2, is unique in acting on both enantiomers of ornithine, although less effectively on the D-enantiomer. We studied the stereospecificity of the enzyme for the hydrogen abstraction from C-5 of the substrate moiety and the addition and removal of the hydrogen at C-4' of the cofactor (pyridoxal phosphate and pyridoxamine phosphate) moiety of the external Schiff base intermediate in the transamination of L- and D-ornithine. L- and D-[5-3H]ornithines were prepared by incubation of L- and D-ornithines with the enzyme in 3H2O, respectively. When the L-[5-3H]ornithine was incubated with L-ornithine 5-aminotransferase of a mesophile, Bacillus sphaericus, which catalyzes the stereospecific abstraction of pro-S hydrogen from C-5 of L-ornithine, most of the tritium was released into the solvent. The D-[5-3H]ornithine also reacted with the enzyme of B. sphaericus in the presence or absence of the amino acid racemase of Pseudomonas putida. Tritium was released only in the presence of the racemase, which catalyzes the racemization of ornithine but does not act on C-5 of ornithine. These results show that the Bacillus sp. YM-2 ornithine 5-aminotransferase stereospecifically abstracts the pro-S hydrogen from C-5 of L- and D-ornithine. When the apo form of the enzyme was incubated with pyridoxamine 5'-phosphate that was stereospecifically tritiated at C-4' and 2-oxoglutarate in the presence of L ornithine or D-ornithine, tritium was released exclusively from (4'S)-[4' 3H]pyridoxamine. Therefore, addition and abstraction of hydrogen at C-4' of the cofactor moiety stereospecifically occur on the si face of the external Schiff base intermediate in the overall transamination catalyzed by Bacillus sp. YM-2 ornithine 5-aminotransferase irrespective of the C-2 configuration of the amino donor. PMID- 8703953 TI - Complementation of coq3 mutant yeast by mitochondrial targeting of the Escherichia coli UbiG polypeptide: evidence that UbiG catalyzes both O methylation steps in ubiquinone biosynthesis. AB - Ubiquinone functions in the mitochondrial electron transport chain. Recent evidence suggests that the reduced form of ubiquinone (ubiquinol) may also function as a lipid soluble antioxidant. The biosynthesis of ubiquinone requires two O-methylation steps. In eukaryotes, the first O-methylation step is carried out by the Coq3 polypeptide, which catalyzes the transfer of a methyl group from S-adenosylmethionine to 3,4-dihydroxy-5-polyprenylbenzoate. In Escherichia coli, 2-polyprenyl-6-hydroxyphenol is the predicted substrate; however, the corresponding O-methyltransferase has not been identified. The second O methylation step in E. coli, the conversion of demethylubiquinone to ubiquinone, is carried out by the UbiG methyltransferase, which is 40% identical in amino acid sequence with the yeast Coq3 methyltransferase. On the basis of the chemical similarity of the first and last methyl-acceptor substrates and the high degree of amino acid sequence identity between Coq3p and UbiG, the ability of UbiG to catalyze both O-methylation steps was investigated. The current study shows that the ubiG gene is able to restore respiration in the yeast coq3 mutant, provided ubiG is modified to contain a mitochondrial leader sequence. The mitochondrial targeting of O-methyltransferase activity is an essential feature of the ability to restore respiration and hence ubiquinone biosynthesis in vivo. In vitro import assays show the mitochondrial leader sequence present on Coq3p functions to direct mitochondrial import of Coq3p in vitro and that processing to the mature form requires a membrane potential. In vitro methyltransferase assays with E. coli cell lysates and synthetically prepared farnesylated-substrate analogs indicate that UbiG methylates both the derivative of the eukaryotic intermediate, 3,4-dihydroxy-5-farnesylbenzoate, as well as that of the E. coli intermediate, 2 farnesyl-6-hydroxyphenol. The data presented indicate that the yeast Coq3 polypeptide is located in the mitochondria and that E. coli UbiG catalyzes both O methylation steps in E. coli. PMID- 8703954 TI - Differential expression and butyrate response of human alkaline phosphatase genes are mediated by upstream DNA elements. AB - Human placentas express high levels of the placental alkaline phosphatase (PLAP) gene and low levels of a highly related gene, germ cell AP (GCAP). Malignant transformation of the placenta is accompanied by a reversal of this pattern of expression. Three Sp1-binding GC-rich DNA elements (sites I-III) located within the first 156 base pairs upstream of the GCAP gene have been shown to direct optimal GCAP gene expression in choriocarcinoma cells. Here we show that the first 100 base pairs upstream of the GCAP gene, which contains sites I and II, constitutes a minimal GCAP promoter. The simultaneous presence of both sites I and II is necessary for GCAP expression and its induction by sodium butyrate. The PLAP promoter directs only a very low level of gene expression in choriocarcinoma cells; the expression does not respond to butyrate. The -100/-1 DNA regions between the GCAP and PLAP promoters differ by only eight base pairs. However, the GC-rich stretches in sites I and II of the GCAP promoter are disrupted in the corresponding PLAP promoter. This disruption blocks or markedly reduces the binding of choriocarcinoma nuclear factors to the PLAP promoter, leading to a reduction in expression and a loss of butyrate response. We further demonstrate that nucleotides -75 to -58 in both AP promoters, which bind a human Y-box binding protein, appear to down-regulate GCAP expression. PMID- 8703955 TI - Enhanced folding of hairpin ribozymes with replaced domains. AB - Reversely joined ribozymes (Komatsu et al., 1995) have been proven to be active. Here we describe the construction of hairpin ribozymes with separated domains, but containing complementary arms for association of the two domains. Linker nucleotides were inserted between the arms and domains. These ribozymes were active under the standard conditions (12 mM MgCl2), depending on the length of the linker. When the complementary arms were covalently joined through a stable loop, these ribozymes showed cleavage activities. However, the K(m) value of the stem-loop ribozymes was found to be larger than that of the parent ribozyme, which can adopt both linear and bent conformations. Kinetic analyses of these modified hairpin ribozymes suggest a higher turnover of the hairpin ribozyme as compared to other small ribozymes. The present ribozymes provide insight into the nature of the domain interaction and are suitable for physicochemical studies on the tertiary structure of the hairpin ribozyme. PMID- 8703956 TI - Probing the protein-DNA contacts of a yeast RNA polymerase III transcription complex in a crude extract: solid phase synthesis of DNA photoaffinity probes containing a novel photoreactive deoxycytidine analog. AB - A novel photoreactive deoxycytidine analog, 4-[N-(p-azidobenzoyl)-2-aminoethyl] dCTP (ABdCTP), has been synthesized and incorporated at specific sites within the SUP4 tRNA(Tyr) gene. Immobilized single-stranded DNA was annealed to specific oligonucleotides and AB-dCMP incorporated into DNA by primer extension. DNA photoaffinity labeling with AB-dCMP was used to survey protein-DNA contacts in initiation and elongation complexes of RNA polymerase III (Pol III), and compared to DNA photoaffinity labeling using the previously described photoreactive deoxyuridine analog, 5-[N-(pazidobenzoyl)-3-aminoallyl]-dUMP (AB-dUMP) [Bartholomew et al. (1993) Mol. Cell.Biol. 13,942-952]. In contrast to previous studies, we have used a crude protein fraction rather than highly purified preparations of Pol III and transcription factors TFIIIC and TFIIIB to examine if some component of the transcription complex is lost upon purification. Eleven nucleotide positions from bp-17 to bp +17 (+1 being the start site of transcription) on the nontranscribed strand were modified and shown to have little or no effect on transcription complex formation, initiation, or elongation as determined by multiple-round transcription assays. Efficient photoaffinity labeling by DNA containing AB-dCMP gave results comparable to that with AB-dUMP at proximal nucleotide positions and provided new evidence for the placement of the 160 and 31 kDa subunits of Pol III near the 5' end of the transcriptional bubble in an elongation complex. A novel 40 kDa protein was cross-linked at bps 17, -9, and -8 in a TFIIIC-dependent manner that had not been previously detected. PMID- 8703957 TI - pH-dependent enhancement of DNA binding by the ultrabithorax homeodomain. AB - Ultrabithorax (Ubx) and Deformed (Dfd) proteins of Drosophila melanogaster contain homeodomains (HD) that are structurally similar and recognize similar DNA sequences, despite functionally distinct genetic regulatory roles for Ubx and Dfd. We report in the present study that Ubx-HD binding to a single optimal target site displayed significantly increased affinity and higher salt concentration dependence at lower pH, while Dfd-HD binding to DNA was unaffected by pH. Results from studies of chimeric Ubx-Dfd homeodomains showed that the N- and C-terminal regions of the Ubx-HD are required for this pH dependence. The increase in binding affinity at lower pH was greater for the Ubx optimal binding site than for other DNA binding sites, indicating that subtle sequence alterations in DNA binding sites may influence pH-dependent behavior. These data demonstrate enhanced DNA binding affinity at lower pH for the Ubx-HD in vitro and suggest the potential for significant discrimination of DNA binding sites in vivo. PMID- 8703958 TI - Steady-state and pre-steady-state kinetic analysis of dNTP insertion opposite 8 oxo-7,8-dihydroguanine by Escherichia coli polymerases I exo- and II exo-. AB - Escherichia coli polymerases (pol) I exo-(KF-) and pol II exo- (pol II-) were used as model enzymes with a DNA primer/template complex (12/16-mer) to examine the kinetics of incorporation of dCTP and dATP at the site of an 8-oxo-7,8 dihydroguanine (8-oxoGua) residue; compared to guanine (Gua). In steady-state assays (with DNA in excess) the rate of incorporation (kcat) was dCTP > dATP and the K(m),dATP < K(m),dCTP during incorporation opposite 8-oxoGua with both polymerases. Pre-steady-state kinetic curves (rapid-quench analysis) for the addition of C opposite 8-oxoGua or Gua by KF- and pol II- were all biphasic, with a rapid initial single-turnover burst followed by a slower multiple turnover rate, while addition of A opposite 8-oxoGua did not display burst kinetics with either enzyme. Reduced rates of incorporation of the dCTP alpha S and dATP alpha S phosphorothioate analogs suggest that the rates of incorporation of A and C opposite 8-oxoGua are limited during polymerization by the rate of phosphodiester bond formation. Neither polymerase appears to discriminate between adducted and nonadducted DNA substrate for binding. Kinetic assays performed with varying dCTP concentrations indicate that dCTP has a higher K(d) and lower k(p) (polymerization rate) for incorporation opposite 8-oxoGua compared to Gua. Furthermore, the dATP binding affinities with KF- and pol II- were approximately 10- and approximately 3-fold lower, respectively, than that of dCTP as determined in competition assays with mixtures of dCTP and dATP. Microscopic rate constants were estimated by mathematical analysis of dNTP concentration dependence curves. Both polymerases preferentially extended the A:8-oxoGua pair while extension of the C:8-oxoGua pair was greatly impaired. Based on these findings, the fidelity of KF- and pol II- during replication of 8-oxoGua depends on contributions from nucleotide binding, the rate of phosphodiester bond formation, and the ease of base pair extension. PMID- 8703959 TI - Solution conformation of the (-)-cis-anti-benzo[a]pyrenyl-dG adduct opposite dC in a DNA duplex: intercalation of the covalently attached BP ring into the helix with base displacement of the modified deoxyguanosine into the major groove. AB - This paper reports on the combined NMR-molecular mechanics computational studies of the solution structure of the (-)-cis-anti-[BP]dG adduct positioned opposite dC in the sequence context d(C1- C2-A3-T4-C5-[BP]G6-C7-T8-A9-C10-C11).d(G12-G13 T14- A15-G16-C17-G18-A19-T20- G21-G22) duplex [designated (-)-cis-anti-[BP]dG.dC 11-mer duplex]. This adduct is derived from cis addition at C10 of (-)-anti 7(S),8(R)-dihydroxy-9(R),10(S)-epoxy-7,8,9,10- tetrahydrobenzo[a]pyrene [(-)-anti BPDE] to the N2 position of dG6 in this duplex sequence. The exchangeable and nonexchangeable protons of the benzo[a]pyrenyl moiety and nucleic acid of the major conformation were assigned following analysis of two-dimensional NMR data sets in H2O and D2O solution. There was a general broadening of proton resonances for a three-nucleotide segment centered about the lesion site which resulted in a tentative assignment for the sugar protons of the C7 residue in the spectrum of the adduct duplex. The solution conformation of the major conformation of the (-) cis-anti-[BP]dG.dC 11-mer duplex has been determined by incorporating DNA-DNA and intermolecular BP-DNA proton-proton distances defined by lower and upper bounds deduced from NOESY data sets as restraints in molecular mechanics computations in torsion angle space. The results establish that the covalently attached benzo[a]pyrenyl ring intercalates between intact Watson-Crick dC5.dG18 and dC7.dG16 base pairs. The modified deoxyguanosine [BP]-dG6 and its partner cytosine dC17 are looped out of the helix into the major groove. The purine ring of the [BP]dG6 residue is directed toward the 5'-end of the modified strand and stacks over the major groove edge of its 5'-side neighbor dC5 residue. The solution structure of the (-)-cis-anti-[BP]dG.dC 11-mer duplex is compared with those of the stereoisomeric (+)-trans-anti-[BP]dG [Cosman, M., et al. (1992) Proc. Natl. Acad. Sci. U.S.A. 89, 1914-1918], (-)-trans-anti-[BP]dG [de los Santos, C., et al. (1992) Biochemistry 31, 5245-5252], and (+)-cis-anti-[BP]dG [Cosman, M., et al. (1993a) Biochemistry 32, 4146-4155] adducts positioned opposite dC in the same duplex sequence context. A key finding is that the long axes of the intercalated benzo[a]pyrenyl rings in the solution structures of the (+)- and (-)- cis-anti-[BP]dG.dC 11-mer duplexes are oriented in opposite directions with the benzylic ring directed toward the minor groove in the (+)-cis isomer and toward the major groove in the (-)-cis isomer. In addition, a comparison is also made with the solution structure of the (+)-trans-anti-[BP]dG adduct opposite a deletion site [Cosman, M., et al. (1994a) Biochemistry 33, 11507-11517] since this adduct duplex displays several conformational features in common with the structure of the (-)-cis-anti-[BP]dG.dC 11-mer duplex. The structures of both duplex adducts exhibit intercalation of the covalently attached ligand into the helix and displacement of the modified deoxyguanosine into the major groove. Studies of the biological activities of stereochemically defined BP-DNA adducts and the comparison of the solution structure of the (-) cis-anti-[BP]dG.dC 11-mer duplex with its stereoisomeric counterparts should lead to new insights into the relationships between defined helical distortions and mutagenic specificity and activity. PMID- 8703960 TI - A fluorescence anisotropy study of DNA binding by HPV-11 E2C protein: a hierarchy of E2-binding sites. AB - Association of the human papillomavirus (HPV) E2 protein with its palindromic DNA binding site is a necessary step for transcriptional trans-activation. To study the interaction between DNA and E2, the carboxyl-terminal domain of HPV-11 E2 protein (E2C) was expressed in Escherichia coli and purified to homogeneity. The binding affinity of the recombinant E2C protein for a single palindromic DNA recognition site was determined using a 5'-fluorescein-labeled 24 base pair oligonucleotide. Competitive titrations between the fluorescein-labeled oligonucleotide and an unlabeled oligonucleotide of identical sequence yielded a native affinity of 4.5 x 10(-9)M. Sequences from the seven E2-binding sites within the HPV-11 genome were titrated to establish a hierarchy of binding site affinities. All high-affinity E2-binding sites are located within or near the HPV 11 LCR. E2-binding sites distant from the LCR appear to have low affinity for E2. When the location and affinity of each E2-binding site are plotted in relation to a transcription map of HPV-11, it is apparent that the major RNA transcripts produced reflect the high-affinity E2-binding sites within the HPV LCR. To assess the E2C-binding contribution of specific base pairs within the oligonucleotide palindrome, additional double-stranded oligonucleotides were prepared in which the central nonpalindromic sequences were varied. While simple strand transposition of the A4.T4 center had a minimal effect upon the E2C oligonucleotide binding affinity, replacement with TATA.ATAT or CGCG.GCGC centers substantially decreased the affinity of E2C for its binding site. Alteration of the canonical portions of the E2-binding palindrome reduced the DNA-protein binding affinity dramatically. PMID- 8703961 TI - Involvement of Phe19 in the Mn(2+)-L-malate binding and the subunit interactions of pigeon liver malic enzyme. AB - A triple mutant, F19S/N250S/L353Q, of pigeon liver malic enzyme was found to have no detectable enzymatic activity [Chou, W.-Y., Huang, S.-M., & Chang, G.-G. (1994) Arch. Biochem. Biophys. 310, 158-166]. In the present study, point mutants at these positions (F19S, N250S, and L353Q) were prepared by site-directed mutagenesis. Both N250S and L353Q have kinetic properties similar to those of the wild-type. On the other hand, the K(m)(app) values for both Mn2+ and L-malate of F19S were increased by approximately 10-fold, while the kcat value was decreased by 5-fold, which results in a decrease of the apparent catalytic efficiency (kcat/K(mNADP)K(mMal)K(mMn) by approximately 300-fold. These results clearly indicate that the F19S mutation is mainly responsible for the undetectable enzyme activity of the triple mutant. Three more Phe19 mutants (F19Y, F19G, and F19A) were then prepared. There is a direct correlation between the size of the substitutes and the affinities for Mn2+ and L-malate. The kinetic parameters for F19Y were similar to those for wild-type. Both F19A and F19G reveal a 5-fold decrease of kcat values. Two K(dMn) values for the high- and low-affinity sites, respectively, were detectable for the wild-type. On the contrary, only one K(dMn) value was detected for the F19 mutants, which was increased in the order of F19G > F19A > F19S > F19Y, with F19G being the most affected mutant. The K(mMal) values of F19G and F19A were increased 100- and 6-fold, respectively. The catalytic efficiency (kcat/K(mNADP)K(dMal)K(dMn)) of F19G was decreased to only 0.01% of that of the wild-type. The above results clearly indicate that the hydrophobic aromatic ring at position 19 plays a critical role in L-malate and Mn2+ binding. Furthermore, all mutants that have a small residue at position 19 exist as monomers. Therefore, Phe19 may locate in or near the regions for Mn(2+) L-malate binding as well as for the subunit contact. These results are compatible with the asymmetric model for the quaternary structure of malic enzyme we proposed previously [Chang, G.-G., Huang, T.-M., Huang, S.-M., & Chou, W.-Y. (1994) Eur. J. Biochem. 225, 1021-1027]. The possible roles of the N-terminus of malic enzyme were also addressed. PMID- 8703962 TI - Metal binding sites of H(+)-ATPase from chloroplast and Bacillus PS3 studied by EPR and pulsed EPR spectroscopy of bound manganese(II). AB - The metal binding sites of isolated F1 ATPase from spinach chloroplasts (CF1) and from the thermophilic bacterium Bacillus PS3 (TF1) have been studied by EPR and pulsed EPR spectroscopy using Mn(II) as a paramagnetic probe. After dialysis in the presence of EDTA, purified CF1 retains 0.14 +/- 0.07 Mg(II) and approximately 0.75 +/- 0.25 ADP. TF1 retains 0.31 +/- 0.03 Mg(II) and 0.08 +/- 0.01 nucleotide (ADP + ATP) after the same treatment. Supplementing known quantities of Mn(II) to metal-depleted CF1 allowed a spectroscopic characterization of the bound Mn(II) cations, for which the EPR spectra at X- and Q-band are reported. The zero field splitting parameters of Mn(II) are derived from the simulation of the EPR signal recorded at Q-band for a sample supplemented with 0.3 Mn/CF1. The values, magnitude of D approximately 200 x 10(-4) cm-1 and magnitude of E approximately 40 x 10(-4) cm-1 suggest that the Mn(II) binds to CF1 in a slightly distorted environment. The ESEEM spectra of complexes of Mn(II) with CF1 were also recorded for different Mn/CF1 ratios. For a complex with 0.8 Mn/CF1, the ESEEM spectrum shows two frequencies at 3.7 and 8.6 MHz that are attributed to the magnetic coupling with 31P with a hyperfine coupling constant of magnitude of A approximately 5.3 MHz, reflecting the interaction with a phosphate group from the endogenous ADP molecule. This demonstrates close proximity of the strong affinity metal site M1 and the endogenous ADP binding site N1, and binding of the ADP beta phosphate to the divalent metal cation. For Mn(II) complexes with higher Mn/CF1 ratios, new frequency components below approximately 5 MHz are resolved in the spectra in addition to the peaks from 31P. From a comparison of the CF1 spectra and their magnetic field dependence across the Mn(II) EPR line shape with those of Mn(II) complexes with imidazole, glycine, poly-L-lysine, and nucleotide ligands, it is concluded that additional metal binding sites are filled at higher Mn contents and that these involve 14N donors. It is suggested that the most probable set of ligands of the divalent metal(s) for these additional metal sites in CF1 includes a lysine residue, in line with a previous proposal [Houseman, A. L. P., Morgan, L., LoBrutto, R., & Frasch, W. D. (1994) Biochemistry 33, 4910 4917]. Similar experiments for a Mn(II) complex with TF1 (0.4 Mn/TF1) showed no interaction with 31P; instead modulations are detected in the ESEEM below approximately 5 MHz that are attributed to a 14N ligand. This is tentatively attributed to the deprotonated amine of Lys-162 from a beta subunit, on the basis of the structural data available for the mitochondrial F1 complex. Addition of the substrate ATP to this Mn.TF1 complex leads to the formation of a ternary Mn.TF1.ATP complex with coordination of the Mn(II) by a phosphate group from the ATP as judged from the ESEEM results (magnitude of A(31P) approximately 4.5 MHz). An increase in the hyperfine coupling constant of 31P of the phosphate bound to Mn(II) to magnitude of A(31P) approximately 5.1 MHz is observed after incubation of the ternary complex at room temperature. This is interpreted as a significant rearrangement of the coordination sphere of the Mn(II) in the M1 site of the Mn.TF1.ATP complex and may reflect conformational changes of catalytic significance that occur in the nucleotide binding site during unisite hydrolysis of ATP to ADP by this complex. PMID- 8703963 TI - Release of lipid vesicle contents by an antibacterial cecropin A-melittin hybrid peptide. AB - A synthetic cecropin A(1-8)-melittin(1-18) hybrid peptide, with antimalarial and antibacterial properties, promotes leakage of aqueous contents of phospholipid vesicles, as determined by measuring the induced release of vesicle-entrapped fluorescence probes. The release of vesicle contents corresponds to an all-or none mechanism. High molecular weight entrapped solutes (fluorescence-labeled dextrans, 20 and 4 kDa molecular mass) are also released by the peptide. This fact and the high peptide stoichiometry required for the release of vesicle contents suggest a detergent-like disruption of the bilayer. The leakage process is not related to any membrane event requiring lipid-mixing between bilayers. The peptide destabilizes both negatively and neutrally charged phospholipid vesicles. The thermal variation of the fluorescence anisotropy of 1,6-diphenyl-1,3,5 hexatriene-labeled vesicles is modified by the peptide. Circular dichroism and tryptophan fluorescence emission spectra reveal conformational changes in the peptide molecule upon interaction with the lipid vesicles. These changes are consistent with an increased alpha-helical content and a less polar environment for the single tryptophan residue of the peptide. The leakage induced in phosphatidylserine vesicles is a faster process than in phosphatidylcholine vesicles, while the peptide is more effective in releasing the contents of the latter type of vesicles. This suggests that acidic phospholipids may modulate the effect of the peptide on membranes. PMID- 8703964 TI - Mechanism of apoptosis suppression by phorbol ester in IL-6-starved murine plasmacytomas: role of PKC modulation and cell cycle. AB - We show here that the mode of cell death in IL-6-starved T1165 and T1198 plasmacytoma cell lines is apoptosis, and that it can be suppressed by phorbol ester (PMA) treatment in a protein kinase C (PKC)-mediated process that involves alpha and/or delta isozymes. PMA-induced PKC activation, but not the depletion that follows it, participates in the suppression of apoptosis. Extended PKC activation is necessary but not sufficient for the apoptosis suppression. In addition, the cells must be in a "competent" state, which appears not to be determined by PKC. We observed two points of "competence" during the time between withdrawal of IL-6 and the start of massive cell death: one, immediately after withdrawal, and another, just before onset of apoptosis, at the time corresponding to maximal accumulation of cells in a G0/G1 block imposed by IL-6 withdrawal. Treatment with PMA and other PKC activators resulted in a shift of the cell population to S phase, lifting the G0/G1 block. We propose a model in which cells are rescued in a certain stage of the G1 phase of cell cycle. Death suppression occurs when a transient PMA-induced PKC activation occurs when a significant number of cells are in this part of G1, allowing them to pass the restriction point safely without initiating the cell death program. PMID- 8703965 TI - The transient kinetics of Escherichia coli chorismate synthase: substrate consumption, product formation, phosphate dissociation, and characterization of a flavin intermediate. AB - Chorismate synthase is the seventh enzyme of the shikimate pathway and catalyzes the conversion of 5-enolpyruvylshikimate 3-phosphate (EPSP) to chorismate. The reaction involves the 1,4-elimination of phosphate and the C-(6proR) hydrogen of the substrate with unusual anti stereochemistry and requires a reduced flavin cofactor. This paper describes the kinetics of the formation and decay of a flavin intermediate, EPSP consumption, chorismate and phosphate formation, and phosphate dissociation during single and multiple turnover experiments, determined using rapid reaction techniques. The kinetics of phosphate dissociation using the substrate analogues (6R)-[6-2H]EPSP and (6S)-6-fluoro-EPSP have also been determined. The observations are consistent with a nonconcerted chorismate synthase reaction. The flavin intermediate is not simply associated with the conversion of substrate to product because it forms before the substrate is consumed. The transient spectral changes must be associated primarily with events such as protonation of the reduced flavin, a charge transfer complex between reduced flavin and an aromatic amino acid, or a conformational change in the protein. This does not rule out the direct role of flavin in catalysis. PMID- 8703966 TI - Lipid regulation of CTP: phosphocholine cytidylyltransferase: electrostatic, hydrophobic, and synergistic interactions of anionic phospholipids and diacylglycerol. AB - The contributions of electrostatic and hydrophobic interactions in the activation of cytidylyl-transferase (CT) by various negatively charged lipids were analyzed using small unilamellar or multilamellar vesicles (SUVs or MLVs). The activation of CT by SUVs containing increasing mole percentages of anionic phospholipids varied in proportion to the net charge associated with the polar head group, suggesting an electrostatic component to the activation. However, increasing ionic strength to neutralize the surface charge enhanced the potency of SUVs containing PA or PG, suggesting that the hydrophobic effect is a stronger force than electrostatics in driving the interaction of CT with SUVs. On the other hand, electrostatics played a more important role in the activation by MLVs. Increasing ionic strength decreased the potency of MLVs containing PG. CT bound to MLVs in the gel state, but was inactive; the enzyme was only active when the MLVs were in the liquid-crystalline state, suggesting an intercalation event. Lowering the pH from 7.4 to 6.2 resulted in a decrease in the negative surface charge required for activation. The binding of CT to PG vesicles was enhanced at acidic pH. The results suggest that at pH 6.2 one or more amino acids on CT that are involved in lipid binding would be protonated. This could enhance the electrostatic effect by increasing the positive charge on CT, or it could enhance the hydrophobic effect by decreasing the negative charge on CT. In addition, maximal activity of CT was decreased at the lower pH, suggesting that active site residues may also be affected. CT was activated by the synergistic interaction of diacylglycerol and anionic phospholipid in SUVs. The synergy between DG and PA at low concentrations suggests the possibility that these second messenger lipids could concertedly regulate CT and thus PC synthesis in response to agonists that stimulate PC hydrolysis via phospholipases C and/or D. PMID- 8703967 TI - Energy and electron transfer upon selective femtosecond excitation of pigments in membranes of Heliobacillus mobilis. AB - Excitation energy transfer steps in membranes of Heliobacillus mobilis were directly monitored by transient absorption spectroscopy with a time resolution of 30 fs under selective excitation within the inhomogeneously broadened bacteriochlorophyll g QY band. The initial anisotropy was found to be > 0.4, indicating that the pigments are excitonically coupled. After initial decay of this anisotropy in < 50 fs, major sub-picosecond components associated with spectral equilibration were identified, corresponding to uphill energy transfer with a 300 fs time constant (812 nm excitation) and downhill energy transfer with 100 and 500 fs components (770 nm excitation). These equilibrations are ascribed predominantly to single excitation transfer steps, as anisotropy measurements showed that equilibration within spectrally similar pigments occurs on the same time scale as spectral equilibration, a situation which contrasts with that in photosystem I. Downhill energy transfer occurs to a significant extent directly to an energetically heterogeneous population of excited states as well as in a sequential way via gradually lower-lying pools of bacteriochlorophyll g. This finding supports a description in which all pigments, including the bluemost absorbing, are spatially organized in a random way rather than in clusters of spectrally similar species. Spectral equilibration is not entirely completed prior to formation of the primary radical pair P798 + A0-, which was found to proceed in a multiexponential way (time constants of 5 and 30 ps). No indication for the formation of radical species other than P798 + A0- on the time scale up to 100 ps was found. PMID- 8703968 TI - Trapping of different lipase conformers in water-restricted environments. AB - Based on a recently reported strategy to rationally activate lipolytic enzymes for use in nonaqueous media [Mingarro, I., et al. (1995) Proc. Natl. Acad. Sci. U.S.A. 92, 3308-3312], we compared the behavior in water-restricted environments of activated vs nonactivated forms of different lipases toward their natural substrates, triacylglycerols. To this end, nine lipases from varied origins (mammalian, fungal, and bacterial) were assayed using simple acidolyses as nonaqueous model reactions. The experimental results for several (though not all) lipases, discussed in the light of current structural and functional information, were collectively consistent with a model where, depending on the "history" of sample preparation, basically two different conformers (open and closed) of the lipase can be trapped (and assayed) in the nonaqueous medium. In particular, for a few prototypic lipases investigated in more detail, the following were shown: (i) the activation strategy permitted them to rationally overcome their reported reluctance to convert saturated, long-chain triglycerides, providing quantifiable nonaqueous rate accelerations of up to 3 orders of magnitude; (ii) the activated conformer exhibited a markedly higher ability than its nonactivated counterpart to bind a ligand (nonhydrolyzable phospholipid) in the nonaqueous medium; and (iii) a clearly distinct selectivity profile toward the substrate chain length was obtained for either conformer. PMID- 8703969 TI - Temperature dependence of endocytosis in renal epithelial cells in culture. AB - Temperature dependence of fluid-phase endocytosis was determined in two renal epithelial cell lines, MDCK cells and LLC-PK1 cells, using Lucifer Yellow or horseradish peroxidase as markers. For both cell lines, grown on solid support as a confluent monolayer, biphasic curves of marker uptake vs. temperature were obtained. The changes in slope occurred around 27 degrees C, a critical temperature at which the lipids of the plasma membrane of MDCK cells enter in the gel state. Activation energies were significantly higher above 27 degrees C (15 22 kcal/mol) than below that critical temperature (9-12 kcal/mol). These data indicate that changes in membrane physical state have marked effects on endocytic processes. They suggest that two mechanisms, with different activation energies are involved in the fluid phase endocytosis by renal epithelial cells in culture. PMID- 8703970 TI - Characterization of the 3' UTR sequence encoded by the AQP-1 gene in human retinal pigment epithelium. AB - The complete 3' UTR sequence encoded by the human aquaporin-1 gene is reported. The sequence encompassed by two cDNA clones showed, 33 nucleotides of 5' UTR sequence, a coding sequence of 807 nucleotides and 1886 nucleotides corresponding to the complete 3' UTR sequence. High similarity with 3' UTR sequences from rat and mouse counterparts was found. Northern blot analysis of several human tissues revealed a 2.8 kbp transcript. These data confirm the existence of water channels in the human retinal pigment epithelium. PMID- 8703971 TI - cDNAs from Onchocerca sp. encoding members of the MRS3/MRS4 class of mitochondrial solute carriers. AB - cDNA clones from the parasitic nematodes Onchocerca volvulus and Onchocerca gibsoni encode homologs of the yeast proteins MRS3 and MRS4. Together with an uncharacterised ORF on chromosome III of Caenorhabditis elegans, these constitute a new class of proteins belonging to the mitochondrial solute carrier protein superfamily. So far, five other members of this protein family have been identified in C. elegans, but levels of identity between these and the Onchocerca proteins were considerably lower. Consideration of cysteine content and overall charge implies that the natural substrates of the nematode proteins are small ions. PMID- 8703972 TI - Biochemical characterization of a V-ATPase of tracheal smooth muscle plasma membrane fraction. AB - A biochemical characterization of a Mg(2+)-ATPase activity associated with a plasma membrane fraction isolated from airway (tracheal) smooth muscle was performed. This enzyme is an integral part of the membrane remaining tightly bound after 0.6 M KCl extraction. This enzyme activity showed a cold inactivation in the presence of ATP and Mg2+. Also, this Mg(2+)-ATPase was stimulated by monovalent anions being Cl-, the best anion for such stimulation, even though Br- and I- were good substitutes and F- was ineffective. This Cl--stimulated activity showed a powerful nucleosidetriphosphatase activity having the following divalent cation specificity: Mg2+ > Mn2+ > Ca2+, where Zn2+ and Fe2+ were ineffective. This ATPase activity was not inhibited by ouabain oligomycin C and vanadate indicating that neither P- or F-ATPases were associated with this enzyme activity. However, the existence of a V-ATPase was shown by the significant inhibition causes by bafilomycin A1. Additionally, this V-ATPase seems to be coupled to Cl- conductor because duramycin inhibited this ATPase activity. The presence of a H+ pump associated to this V-ATPase was shown indirectly, through the stimulatory effect produced by uncouplers such as FCCP and 1799, which were able to produce significant stimulation of this V-ATPase indicating the existence of a H(+)-ATPase. Finally, the immunodetection of a 72 kDa polypeptide using a specific antibody against the A subunit (72 kDa) of V-ATPase from chromaffin granule demonstrated the presence of a V-ATPase in this plasma membrane fraction. PMID- 8703973 TI - H+, K+, and Na+ transport across phospholipid vesicular membrane by the combined action of proton uncoupler 2,4-dinitrophenol and valinomycin. AB - The decay of the pH difference (delta pH) across soyabean phospholipid vesicular membrane (created by temperature jump), by the combined action of valinomycin and 2,4-dinitrophenol (DNP) has been monitored with the help of fluorescence from pyranine entrapped inside the vesicles under a variety of concentration conditions. The results suggest the following for the pH region of our interest (pH approximately 6 to pH approximately 8): (i) The rate limiting step in the proton transport cycle is not the transport of proton as DNPH, but the back transport of DNP- and the alkali metal ion M+ as Val-M(+)-DNP- across the membrane. The rate constant associated with the transport of the ternary complex has been estimated to be approximately 1.5 x 10(3) s-1. (ii) The dissociation constant of the ternary complex Val-M(+)-DNP- in the membrane are approximately 1 mM for M+ = K+ and approximately 0.001 mM for M+ = Na+. (iii) The reduction in the cation selectivity of valinomycin on complexing with DNP- is much more than that observed with the anionic form of carbonyl cyanide m-chlorophenylhydrazone (CCCP). The results also provide a verification of a corollary of Mitchell's hypothesis: an experimental strategy which enhances the delta pH decay rate should also be a strategy for the efficient uncoupling of oxidative and photophosphorylation. PMID- 8703974 TI - The mechanism of L-glutamate transport by lactating rat mammary tissue. AB - The transport of L-glutamate by lactating rat mammary gland has been examined using both tissue explants and a perfused mammary preparation. L-Glutamate uptake by mammary tissue explants was predominantly via a Na(+)-dependent pathway: Li+, choline+ and NMDG+ could not substitute for Na+. L-Glutamate efflux from preloaded explants was also influenced by the transmembrane Na(+)-gradient. These results are consistent with (Na(+)-glutamate) cotransport. The Na(+)-dependent system for L-glutamate transport in tissue explants was saturable (Km = 112.5 +/- 19.7 microM; Vmax = 71.3 +/- 10.4 nmol/min per g cells) and selective for anionic amino acids. Thus, D- and L-aspartate were high affinity inhibitors of L glutamate uptake whereas neutral amino acids were relatively ineffective. D Aspartate inhibited L-glutamate uptake in a competitive fashion. L-Glutamate uptake by the perfused mammary gland was (a) Na(+)-dependent (b) saturable (Km = 18.1 +/- 4.9 microM; Vmax = 40.3 +/- 3.7 nmol/min per g tissue) and (c) selective for anionic amino acids. The results suggest that the (Na(+)-glutamate) cotransporter is situated in the blood-facing aspect of the mammary epithelium. PMID- 8703975 TI - Characteristics of Na+/K(+)-ATPase mediated proton current in Na(+)- and K(+) free extracellular solutions. Indications for kinetic similarities between H+/K(+)-ATPase and Na+/K(+)-ATPase. AB - The Na+/K(+)-ATPase of an ouabain-resistant mutant of Torpedo californica and of rat was expressed in Xenopus oocytes by microinjection of mRNA coding for the alpha- and the beta-subunit of the protein. Electrophysiological measurements were performed by means of the extracellular giant-patch clamp technique. The pump currents were analyzed in nominal absence of extracellular Na+ and K+ ions. Under these conditions strongly inward rectifying, ouabain-sensitive current was detected with reversal potentials depending on extracellular pH. Presence or absence of intracellular Na+ or K+ ions had nearly no effect on the inward currents, removal of intracellular ATP caused their inhibition. The reversal potentials of the currents generated by the rat pump was shifted by 82.7 +/- 5.4 mV per 10-fold increase of extracellular proton concentration. This refers to an effective valency of 0.71 +/- 0.05. In the absence of a transmembrane proton gradient the pump current reversed at -64.2 +/- 4.4 mV. These findings are not compatible with a proton conducting channel formed by the pump molecule (Wang and Horisberger (1995) Am. J. Physiol. CP 37, C590-595). Therefore, a kinetic model based on the Post-Albers scheme with a modification derived from the reaction scheme of the gastric H+/K(+)-ATPase is proposed. Together with voltage-dependent binding of extracellular protons, this model is compatible with the observe pump currents. PMID- 8703976 TI - Isolation of mitochondrial porin of the fly Protophormia: porin modification by the pesticide CGA 140'408 studied in lipid bilayer membranes. AB - Mitochondrial porin from the fly Protophormia was solubilized with detergent from whole mitochondria and purified by chromatography across a hydroxyapatite (HPT) column. The purified protein had an apparent molecular mass of about 30 kDa on SDS-PAGE. Partial sequencing of the protein confirmed that it is porin. When reconstituted in planar lipid bilayer membranes, porin formed ion-permeable channels with single-channel conductances of 2.4 and 4.5 nS in 1 M KCl. At low voltage, Protophormia porin displayed the properties of a general diffusion pore and had a small selectivity for anions over cations. At transmembrane potentials starting with about 20-30 mV, the channel switched in closed state, which is still ion-permeable. Our results suggest that Protophormia porin possesses functional properties similar to those of other mitochondrial porins. Porin was also isolated and purified from mitochondria, which were treated with the carbodiimide CGA 140'408 It represents the active derivative of diafenthiuron a new acaricide and insecticide. This carbodiimide labels both a F0-component of the inner membrane ATPase and outer membrane porin in a similar way as N,N' dicyclohexylcarbodiimide (DCCD). Reconstitution experiments with the CGA 140'408 modified porin showed no significant effect of the modification on the single channel conductance, suggesting that CGA 140'408 binds outside the channel. The voltage-dependence of the CGA 140'408-modified porin was changed with respect to the unmodified form. The closed configuration of the pesticide-modified channel was reached at smaller transmembrane potentials, suggesting a shift of the open to the closed state of Protophormia porin by pesticide binding. A possible contribution of this effect to the pesticide action is discussed. PMID- 8703977 TI - Uptake of neutral alpha- and beta-amino acids by human proximal tubular cells. AB - The transport characteristics of amino acids in primary cell cultures from the proximal tubule of human adults (AHKE cells) were examined, using alpha aminoisobutyric acid (AIB) and beta-alanine as representatives of alpha- and beta amino acids, respectively. The Na(+)-gradient dependent influx of AIB occurred by a single, saturable transport system, whereas the Na(+)-gradient dependent uptake data for beta-alanine could be described in terms of two-independent transport components as well as one-transport one-leak model with identical kinetic constants for the high-affinity system. Competition experiments revealed that all the neutral amino acids tested reduced the uptake of AIB, whereas there was no effect of taurine, L-aspartic acid, and L-arginine. By contrast, the influx of beta-alanine was only drastically reduced by beta-amino acids, whereas the inhibition by neutral alpha-amino acids was relatively low. Nor did L-arginine and L-aspartic acid affect the uptake of beta-alanine into AHKE cells. Comparison with the results obtained for normal (NHKE) and immortalized (IHKE) embryonic cells suggested an unaltered expression of the types of transport carriers for neutral alpha- and beta-amino acids in the embryonic and AHKE cells. However, the uptake capacity of the above-mentioned transport proteins was relatively smaller in the embryonic kidney compared with the adult human kidney, which may explain, at least partly, the phenomenon of physiologic amino aciduria in neonates. PMID- 8703978 TI - Membrane fluidity response to odorants as seen by 2H-NMR and infrared spectroscopy. AB - Fourier transform infrared spectroscopy (FTIR) and deuterium nuclear magnetic resonance spectroscopy (2H-NMR) have been used to study the location of two odorants, beta-ionone and menthone, in a model membrane of dimyristoylphosphatidylcholine, as well as the effect of the odorants on the structure and dynamics of the phospholipids. The interaction has been investigated for two lipid-to-odorant molar ratios, 10:1 and 1:1. The two odorants were found to affect the fluidity of the membrane. More specifically, the 2H-NMR results indicate that at a lipid-to-odorant molar ratio of 10:1, both beta-ionone and menthone increase the order of the deuterons in the interfacial and headgroup regions of the lipid while the incorporation of the odorants at a lipid-to-odorant molar ratio of 1:1 decreases the order of both the lipid headgroup and acyl chains. On the other hand, the infrared results show that the incorporation of beta-ionone and menthone decreases the phase transition temperature and cooperativity of the lipid acyl chains. The results suggest that the site of incorporation of beta-ionone and menthone is very similar in DMPC membranes. PMID- 8703979 TI - Alteration of Na(+)-coupled transport in site-directed mutants of the melibiose carrier of Escherichia coli. AB - Asn-58 of the Escherichia coli melibiose carrier was replaced by Ala, Leu, Ser, and Gln. Trp-54 was replaced by Leu and a double mutant Leu-54/Ala-58 was constructed using site-directed mutagenesis. Cation/sugar cotransport and sugar induced cation uptake were studied for each mutant. The change of Asn-58 to Ala results in a nearly complete loss of Na(+)-stimulated galactoside transport as well as sugar-stimulated Na+ uptake. Substitutions of Leu, Gln, and Ser for Asn 58 were also defective in Na(+)-stimulated sugar transport. The Trp-54 to Leu mutant shows moderate sugar accumulation with cation selectivity similar to wild type. The double mutant Leu-54/Ala-58 shows elevated H(+)-melibiose cotransport as well as reduced Na(+)-stimulated melibiose cotransport. These results suggest that Asn-58 is important for Na+ recognition. PMID- 8703980 TI - Receptor affinity of neurotensin message segment immobilized on liposome. AB - Neurotensin derivatives having a dioctadecyl group were synthesized and immobilized on DMPC liposome to construct a multivalent-ligand system. The derivatives are Ac-Glu[N(C18H37)2]-(Sar-Sar-Pro)n-Arg-Arg-Pro-Tyr-Ile-Leu-OH (D3nNT, n = 0,1,2,3), where a dioctadecyl group was connected to the N-terminal side of neurotensin 8-13 fragment directly or through a hydrophilic and flexible spacer chain of different lengths. The derivatives were spontaneously immobilized on DMPC liposome upon incubation overnight. The receptor affinity of the derivatives increased significantly upon immobilization on liposome. The maximum affinity was obtained by D9NT immobilized on DMPC liposome at the molar ratio of DMPC and D9NT of 200. This affinity is slightly better than the neurotensin 8-13 fragment, the message segment of the derivatives. The fluorescent microscopy using rhodamine-labelled liposome revealed that the multivalent-ligand system binds to specific receptors without dissociation of the derivative from DMPC liposome. PMID- 8703981 TI - Human placental ATP diphosphohydrolase is a highly N-glycosylated plasma membrane enzyme. AB - Human placental ATP diphosphohydrolase (ATP-DPH), has been previously characterized as an azide-sensitive, Ca(2+)- or Mg(2+)-dependent triphospho- and diphosphonucleosidase which migrates as an 82 kDa protein band on SDS-PAGE (Christoforidis, S. et al. (1995) Eur. J. Biochem. 234, 66-74). In this paper we have studied the subcellular localization of placental ATP-DPH by differential centrifugation and flotation experiments. Using specific enzymatic markers it was found that ATP-DPH is localized on plasma membrane. ATP-DPH was found to be a highly N-glycosylated protein which is a common post-translational modification of plasma membrane proteins. Extensive incubation of the native pure enzyme with N-glycosidase F resulted in the elimination of the 82 kDa form and the concurrent formation of a deglycosylated product of 57.5 kDa and four other intermediate products, indicating the presence of at least five N-glycosylation sites within the ATP-DPH molecule. The partially deglycosylated sample retained its activity in solution and in native gel electrophoresis and activity staining. PMID- 8703982 TI - Pathways for flip-flop of mono- and di-anionic phospholipids in the erythrocyte membrane. AB - The inward translocations (flip), from the outer to the inner membrane leaflet of human erythrocytes, of di-anionic NBD-labeled phospholipids containing as a head group phosphate esters of glycolate, butyrate and hydroxyethanesulfonate are slow processes (k = 0.005-0.008 h-1, 37 degrees C) at pH 7.4. A decrease of pH highly stimulates the flip. A major role of the anion exchanger (AE1), band 3, in this flip is indicated by (a) the strong inhibition of the flip (55-85%) by stilbene disulfonates and other inhibitors of anion transport, (b) the stimulation and loss of pH dependence of the flip after modification of band 3 by Woodward's reagent K and NaBH4, and (c) the stimulation of the flip after proteolytic cleavage of band 3 by papain. The flip of mono-anionic NBD-phospholipids with phosphate esters of glycerol, glycol, methanol, butanol and benzyl alcohol is much faster than that of their dianionic analogs (k = 0.04 to > 3.0 h-1, 37 degrees C). It is inhibited by stilbene disulfonates to a decreasing extent (35 to 0%) and is not affected by several reversible inhibitors of anion exchange. This indicates a minor component of band-3-mediated flip and a major component of nonmediated flip. The outward translocations (flop), from the inner to outer membrane leaflet, of both mono- and di-anionic phospholipids are very fast (1.0 5.9 h-1), ATP-dependent and inhibitable by vanadate, fluoride, SH-reagents or Mg(2+)-depletion of cells and thereby likely to be largely mediated by a 'floppase'. The stationary distributions of the NBD-labeled anionic phospholipids are asymmetric to an extent (outer to inner leaflet ratio 2-9) correlating with the ratio of the rates of the outward and the inward translocation. Thus, asymmetry is largely abolished by blockage of the floppase-mediated translocation. PMID- 8703983 TI - Sodium-dependent high-affinity binding of carnitine to human placental brush border membranes. AB - The interaction of carnitine with human placental brush-border membrane vesicles was investigated. Carnitine was found to associate with the membrane vesicles in a Na(+)-dependent manner. The time course of this association did not exhibit an overshoot, which is typical of a Na+ gradient-driven transport process. The absolute requirement for Na+ was noticeable whether the association of carnitine with the vesicles was measured with a short time incubation or under equilibrium conditions, indicating Na(+)-dependent binding of carnitine to the human placental brush-border membranes. The binding was saturable and was of a high affinity type with a dissociation constant of 1.37 +/- 0.03 microM. Anions had little or no influence on the binding process. The binding process was specific for carnitine and its acyl derivatives. Betaine also competed for the binding process, but other structurally related compounds did not. Kinetic analyses revealed that Na+ increased the affinity of the binding process for carnitine and the Na+/carnitine coupling ratio for the binding process was 1. The dissociation constant for the interaction of Na+ with the binding of carnitine was 24 +/- 4 mM. This constitutes the first report on the identification of Na(+)-dependent high-affinity carnitine binding in the plasma membrane of a mammalian cell. Studies with purified rat renal brush-border membrane vesicles demonstrated the presence of Na+ gradient-driven carnitine transport but no Na(+)-dependent carnitine binding in these membrane vesicles. In contrast, purified intestinal brush-border membrane vesicles posses neither Na+ gradient-driven carnitine transport nor Na(+)-dependent carnitine binding. PMID- 8703984 TI - Vesicles made of glycophospholipids with homogeneous (two fluorocarbon or two hydrocarbon) or heterogeneous (one fluorocarbon and one hydrocarbon) hydrophobic double chains. AB - The vesicle-forming ability of the new anionic double chain glycophospholipids 1 4, with either two hydrocarbon or two perfluorocarbon chains, or a mixed double chain (one fluorinated, one hydrogenated), was investigated. When dispersed in water, 1a-c,e, 2b,c and 4b,c readily gave heat-sterilizable vesicles, 30-70 nm in diameter. The galactose and mannose-based fluorinated vesicles were also highly stable on aging. The 6-substituted glucose derivatives 3 formed tubules that reversibly interconverted into vesicles, depending on temperature. The leakage rate in buffer of carboxyfluorescein or calcein from vesicles made from 1a-c,e 2b,c and 4b,c depended on the sugar (t1/2 galactose > mannose > glucose). It decreased significantly with increasing fluorination and length of the hydrophobic tails. The mixed perfluorocarbon/hydrocarbon-tailed amphiphiles were found to be miscible with both the two fluorocarbon chains and the two hydrocarbon chains derivatives. Such admixing tended, however, to increase the small unilamellar vesicles' permeability. In buffered serum, all the vesicles investigated were highly permeable, but incorporation of cholesterol or DSPC in vesicles made of 1e significantly reduced their permeability in serum. The new vesicle and membrane components have i.v. maximum tolerated doses as high as 500 mg/kg body weight in mice; hemolytic activity sharply decreases with increasing degree of fluorination. PMID- 8703985 TI - Monovalent cations differentially affect membrane surface properties and membrane curvature, as revealed by fluorescent probes and dynamic light scattering. AB - The effects of monovalent cations on the interfacial electrostatic potential (psi d), hydrodynamic shear boundary distance (ds), and membrane curvature were studied in large unilamellar phospholipid and galacto/sulfolipid liposomes containing different fractions of negatively charged lipids. The differential effects of alkali metal ions on psi d could be accurately determined at physiological surface charge densities with a surface-anchored fluorescent probe. Li+ and Na+ more effectively decrease psi d and exhibit higher association constants (Kas) than K+ and Cs+. These two groups of cations display qualitatively different perturbations of the interfacial structure. Combining Kas values with the electrokinetic (zeta) potentials yielded the respective ds values. At low ionic strength ds more substantially increases with Li+ or Na+ than with K+ or Cs+. Increasing surface charge density causes increased membrane curvature in the presence of K+ or Cs+, but this is largely prevented by Li+ or Na+. Membrane binding of the amphiphilic cation acridine orange decreases surface charge and membrane curvature more extensively than H3O+, Li+, and Na+. The differential interface-perturbing behavior of monovalent cations is discussed with regard to their different hydration tendencies that will modulate the extent and stability of the hydrogen-bond network along the charged membrane surface. PMID- 8703986 TI - Structural requirements for intracellular transport of pulmonary surfactant protein B (SP-B). AB - Human SP-B is synthesized by the alveolar Type II epithelial cell as a 381 amino acid preproprotein. The 79 residue mature SP-B peptide is extremely hydrophobic and flanked by propeptides of 200 and 102 amino acids at its NH2- and COOH termini, respectively. The purpose of this study was to identify peptide domains of the SP-B proprotein necessary for trafficking of the mature peptide in the secretory pathway. To this end several constructs were generated, by subcloning the full length human SP-B (SP-B), COOH-terminally truncated SP-B (SP-B delta C, in which residues 201-381 were deleted), NH2-terminally deleted SP-B (SP-B delta N, in which residues 28-200 were deleted), NH2-terminal propeptide (SP-BN), mature SP-B (SP-BM) and COOH-terminal propeptide (SP-BC), into the mammalian expression vector pcDNA3. The resulting expression constructs were characterized by DNA sequencing and in vitro transcription/translation and subsequently transfected into Chinese hamster ovary cells. 48 h after transfection, cells were labeled with [35S]-met/cys and analyzed by immunoprecipitation, SDS-PAGE and autoradiography. Proteins encoded by SP-B, SP-B delta C, SP-BN and SP-BC constructs were secreted into media; in contrast, SP-B constructs lacking the NH2 terminal propeptide (SP-B delta N) remained in the endoplasmic reticulum (as assessed by endoglycosidase H sensitivity) and were rapidly degraded. We conclude that (1) 27 amino acids at the NH2-terminus of SP-B contain a functional signal peptide and (2) the NH2-terminal propeptide of the SP-B precursor is necessary and sufficient for intracellular trafficking of the mature peptide. PMID- 8703987 TI - Human immunodeficiency virus type-1 (HIV-1) infection increases the sensitivity of macrophages and THP-1 cells to cytotoxicity by cationic liposomes. AB - Cationic liposomes may be valuable for the delivery of anti-sense oligonucleotides, ribozymes, and therapeutic genes into human immunodeficiency virus type 1 (HIV-1)-infected and uninfected cells. We evaluated the toxicity of three cationic liposomal preparations, Lipofectamine, Lipofectin, and 1, 2 dimyristyloxypropyl-3-dimethyl-hydroxyethyl ammonium bromide (DMRIE) reagent, to HIV-infected and uninfected cells. Monocyte/macrophages were infected with HIV 1BaL and treated with liposomes in medium containing 20% fetal bovine serum (FBS) for 4 h or 24 h at 37 degree C. Uninfected monocytic THP-1 cells and chronically infected THP-1/HIV-1IIIB cells were treated with phorbol 12-myristate 13-acetate (PMA) and exposed to liposomes in the presence of 10% FBS. Toxicity was evaluated by the Alamar Blue assay and viral p24 production. The toxic effect of cationic liposomes was very limited with uninfected cells, although concentrations of liposomes that were not toxic within a few days of treatment could cause toxicity at later times. In HIV-1BaL-infected macrophages, Lipofectamine (up to 8 microM) and Lipofectin (up to 40 microM) were not toxic after a 4-h treatment, while DMRIE reagent at 40 microM was toxic. While a 4-h treatment of THP-1/HIV-1IIIB cells with the cationic liposomes was not toxic, even up to 14 days post treatment, all three cationic liposomes were toxic to cells at the highest concentration tested after a 24-h treatment. Similar results were obtained with the Alamar Blue assay, Trypan Blue exclusion and a method that enumerates nuclei. Infected cells with relatively high overall viability could be impaired in their ability to produce virions, indicating that virus production appears to be more sensitive to treatment with the cationic liposomes than cell viability. Our results indicate that HIV-infected cells are more susceptible than uninfected cells to killing by cationic liposomes. The molecular basis of this differential effect is unknown; it is proposed that alterations in cellular membranes during virus budding cause enhanced interactions between cationic liposomes and cellular membranes. PMID- 8703988 TI - Biological activity of 26-succinylbryostatin 1. AB - Bryostatin 1, a macrocyclic lactone, has undergone phase I trials as an anticancer agent. Because of the lipid solubility of this compound it must be delivered either in ethanol or in a PET formulation. During the trial, these vehicles caused a large number of treatment-related side effects. We have synthesized the triethanolamine salt of 26-succinylbryostatin 1 and find that this compound is approx. 100-fold more water soluble than bryostatin 1. Because of the potential for clinical use, we have evaluated the biologic activity of this compound. We find that in a concentration-dependent manner 26 succinylbryostatin 1 is capable of activating protein kinase C (PKC) in vitro and displacing [3H]PDBu from PKC. However, at all concentrations tested the activity was less than the parent compound bryostatin 1. Addition of bryostatin 1 but not 26-succinylbryostatin 1 to U937 leukemic cells in culture stimulated a drop in cytosolic PKC, secondary to translocation of PKC to the membrane. Although 26 succinylbryostatin 1 did not stimulate a drop in the cytosolic levels of PKC, addition to U937 cells activated transcription from an AP-1 enhancer construct and c-Jun protein phosphorylation in a similar fashion to bryostatin 1 and differentiation of U937 cells. Unlike bryostatin 1, 26-succinylbryostatin 1 was unable to cause aggregation of human platelets. Although injection of bryostatin 1 into mice carrying B16 melanoma inhibits tumor growth, there was no significant inhibition of melanoma growth when identical doses of 26-succinylbryostatin 1 were injected. Therefore, 26-succinylbryostatin 1 shares some but not all of the pharmacologic properities of bryostatin 1. This compound can activate protein phosphorylation without lowering cytosolic levels of PKC. PMID- 8703989 TI - Controlled overexpression of selected domains of the P85 subunit of phosphatidylinositol 3-kinase reverts v-Ha-Ras transformation. AB - Selected domains of the regulatory p85 subunit of phosphatidylinositol 3-kinase have been expressed under the control of the tetracycline transactivator in NIH 3T3 fibroblasts transformed by the v-Ha-Ras oncogene. The domains expressed were the SH3 domain, the BCR homology domain, the region between the two SH2 domains which contains the p110 binding site (the inter SH2 (IS) domain), and the C terminal (CT) domain (containing both SH2 domains and the IS domain). The levels of IS or SH3 domain expressed in the presence of tetracycline were sufficient to reverse the transforming effects of v-Ha-Ras, and no further inhibition of proliferation was observed when expression was increased 7-fold by removal of tetracycline. In contrast inhibition of proliferation by the CT domain was observed only when the level of expression was increased 5-fold by removal of tetracycline. Overexpression of the BCR domain of p85 had no effect on v-Ha-Ras transformation. Expression of the IS domain disrupted the interaction of the p85 regulatory subunit with the p110 catalytic subunit. These results indicate that the association of the p85 subunit of PI 3-kinase with the p110 subunit is necessary for v-Ha-Ras-induced transformation in NIH 3T3 cells. PMID- 8703990 TI - The plasma membrane NADH-diaphorase is active during selective phases of the cell cycle in mouse neuroblastoma cell line NB41A3. Its relation to cell growth and differentiation. AB - Plasma membrane oxidoreductases have been described in all cells and use extracellular impermeant electron acceptors (DCIP, Ferricyanide) that are reduced by NADH. They appear to regulate the overall cell activity in response to oxidative stress from the cellular environment. An NADH-DCIP reductase has been described at the plasma membrane of NB41A3, a neuroblastoma cell line (Zurbriggen and Dryer (1993) Biochim. Biophys. Acta 1183, 513-520) whose activation with extracellular impermeant substrates promotes cell growth. Elutriation was performed to separate cells and the various fractions were analysed for enzyme activity on intact cells combined with flow cytometry. These studies showed that the enzyme is mostly induced and activated during the G1 and during the G2/M phases. These observations were further corroborated with specific inhibitors of the cell cycle. A three-fold increase in enzyme activity was observed in the presence of alpha-amanitin, a specific cell cycle inhibitor of the G1-phase. Taxol, a specific inhibitor of the M-phase, also induces a significant increase in enzyme activity. FACS analysis of taxol -treated and alpha-amanitin-treated cells corroborated these data. The cells have been synchronized and the enzyme activity was measured at different time intervals. An activity increase was observed after ca. 2-3 h, that corresponds to a raise in the M-phase, according to FACS data. Furthermore, NTera-2 cells - a human neuroblastoma cell line that differentiates into fully mature neurones in the presence of retinoic acid - exhibit a 50% decrease in the enzyme activity during the G0-phase upon differentiation, compared to undifferentiated cells. Together the data presented in this paper show that this plasma membrane NADH-diaphorase affects cell growth and differentiation and is strongly modulated at various phases of the cell cycle. PMID- 8703991 TI - Phenylarsine oxide and vanadate: apparent paradox of inhibition of protein phosphotyrosine phosphatases in rat adipocytes. AB - Vanadate mimics, whereas phenylarsine oxide (PAO) antagonizes, the effects of insulin in rat adipocytes. Both vanadate and PAO are documented inhibitors of protein-phosphotyrosine phosphatases. The relationship between the inhibition of 'inhibitory' PTPase and 'stimulatory' PTPase has been studied here in primary rat adipocytes. Low concentrations of PAO (IC50 = 0.6-2.0 microM) blocked the stimulating effects of insulin, vanadate and pervanadate on hexose uptake and glucose metabolism. Inhibition of isoproterenol-mediating lipolysis by vanadate and insulin was not blocked by PAO. The activating effects of okadaic acid on hexose uptake and glucose metabolism, which occur at points downstream to tyrosine phosphorylation, were also not blocked by PAO. Subsequent studies suggested that the PAO-sensitive PTPase comprises a minute fraction of the total adipocytic PTPase activity. To identify its location we applied procedures involving fractionations and activation of non-receptor adipocytic protein tyrosine kinase by PAO and vanadate in cell free assays. We found that the 'inhibitory' PTPase is exclusively associated with the membrane fraction whereas the 'stimulatory' PTPases are present in both the cytosolic and plasma membrane compartments. We next searched for markers, possibly associated with PAO dependent desensitization and found that several proteins became phosphorylated on tyrosine moieties in the supernatant of PAO but not in vanadate pretreated adipocytes. In summary, we propose the presence of a minute, plasma membrane associated PTPase in primary rat adipocytes, inhibition of which arrests the activation of glucose metabolism. In contrast, inhibition of all the other cellular adipose PTPases, ultimately activates rather than inhibits these same bioeffects. PMID- 8703992 TI - Expression of the human RAD51 gene during the cell cycle in primary human peripheral blood lymphocytes. AB - The S. cerevisiae RAD51 gene product exerts important functions in meiotic and mitotic recombination, as well as in the repair of DNA double-strand breaks. We have studied the expression of the human RAD51 (HsRAD51) gene in primary human peripheral blood lymphocytes (PBLs). The HsRAD51 mRNA level increased three fold in mitogen stimulated PBLs, with a peak in the late S phase. A five fold increase of HsRAD51 protein levels was observed in late G2. Specific inhibition of DNA synthesis with aphidicolin did not block the induction of the HsRAD51 protein, indicating that HsRAD51 expression is independent of DNA replication. In contrast, after inhibition of RNA synthesis with actinomycin D and protein synthesis with cycloheximide, the HsRAD51 protein level decreased rapidly. Taken together, these results indicate that the HsRAD51 gene is transcriptionally regulated in human PBLs, and exerts its function during the late S and G2 phases of the cell cycle. PMID- 8703993 TI - Measurement of endosome pH following folate receptor-mediated endocytosis. AB - Free folic acid is believed to enter some cells by folate receptor-mediated endocytosis at membrane invaginations termed caveolae. Folate conjugated macromolecules also enter cells by folate receptor-mediated endocytosis, but their site of entry has never been conclusively identified. In this paper, we show that internalization of folate-macromolecule conjugates by receptor-bearing KB cells can be blocked by agents that specifically inhibit caveolae assembly or internalization such as nystatin and phorbol-12-myristate acetate (PMA). To characterize the intracellular conditions to which the macromolecule-folate conjugates are subsequently exposed, we have measured the pH of the major compartments of the folate endocytosis pathway. pH values of individual endosomal compartments in KB cells were determined by dual-excitation laser-scanning confocal microscopy, where the fluorescence ratio of folate-DM-NERF-dextran (pH sensitive) and Texas Red-dextran (pH-insensitive) was used to calculate pH. These studies revealed that the pH of folate conjugate-containing endosomes commonly varies between 4.7 and 5.8, with the pH in some endosomes as low as 4.3. The most frequent pH value in these compartments was approximately 5.0. PMID- 8703994 TI - Structure and expression of the promoter for the R4/ALK5 human type I transforming growth factor-beta receptor: regulation by TGF-beta. AB - The type I transforming growth factor-beta (TGF-beta) receptors are serine/threonine kinases that are essential for the action of TGF-beta. In this paper, we describe the molecular cloning and expression of the R4/ALK5 human type I TGF-beta receptor promoter. DNA sequence analysis indicates that the promoter lacks a TATA and CAAT box but is highly GC-rich and contains putative Sp1 binding sites. The transcriptional start site is approx. 232 base pairs upstream of the AUG start codon. In human lung fibroblasts, TGF-beta induced a 3-fold increase in steady-state level for type I receptor mRNA. Exposure of cells transfected with a 618 bp promoter fragment to TGF-beta 1 up-regulated transcriptional activity indicating that a TGF-beta response element is contained within this region. PMID- 8703996 TI - Phorbol ester-induced shedding of intercellular adhesion molecule-1 (ICAM-1) on erythroleukemic K 562 cells. AB - The effect of phorbol 12-myristate 13-acetate (PMA) on the expression and shedding of intercellular adhesion molecule-1 (ICAM-1) was investigated on the hematopoietic cell lines K 562 and U 937 using flow cytometry, fluorescence microscopy and ELISA technique. At low concentration of 1 nM, PMA stimulated the expression of ICAM-1 on the cell surface about 4-fold within 24 h, whereas a short-term treatment with 100 nM PMA led to the shedding of 35% of ICAM-1 from the surface of K 562 cells. The release of surface ICAM-1 was found on single cells by fluorescence microscopy to be a uniform process proceeding within 15 min. The shedding of ICAM-1 correlated with elevated levels of sICAM-1 in the supernatants of cultured cells. Also on K 562 cells stimulated by TNF-alpha, a PMA-induced release of ICAM-1 was observed in addition to the known spontaneous shedding. In contrast to the results with K 562 cells, no PMA-induced shedding of ICAM-1 was found on U 937 cells. This indicates a cell-specific process for K 562 cells. The PMA-mediated release of ICAM-1 from K 562 cells suggests that the shedding process does not only occur in parallel to the surface expression of ICAM-1, but may be controlled by particular mechanisms of down-regulation. PMID- 8703997 TI - Impact of tobacco-free policy on recruitment and retention of adolescents in residential substance abuse treatment. AB - As residential treatment programs consider adopting smoke-free policies, treatment providers question whether such policies interfere with patient recruitment and retention. This study assesses the effect of a smoke-free policy on retention using a sample of 155 low-income, mainly minority youth, randomly assigned to two long-term adolescent residential programs at the New Jersey Substance Abuse Treatment Campus. One of these programs has a smoke-free policy, the other does not. Smoking rates are high for the sample. Eighty-five percent of those adolescents smoke, on average half a pack daily, and 39% smoke a pack or more daily. Differential dropout rates between programs for key time periods related to tobacco withdrawal were analyzed to assess the potential effect of smoking policy on retention. Differential dropout rates at the point of program assignment, in the first two days of residency (p = .43), and for the first two weeks of residency (p = .37) show no differences between programs. While analyses showed heavy smokers are more likely than others to leave treatment early, they were likely to leave from either program regardless of smoking policy. As well, verbal expressions of dissatisfaction with smoking policy did not result in drop outs. These results indicate that smoke-free policies have no detrimental effect on program retention. PMID- 8703995 TI - Characterization of adenylyl cyclase stimulated by VIP in rat and mouse peritoneal macrophage membranes. AB - Vasoactive intestinal peptide (VIP) stimulated adenylyl cyclase activity in rat and mouse peritoneal macrophage membranes. GTP potentiated the stimulatory effect of VIP so that it was routinely included at 10 microM GTP. Other agents like GTP, Gpp(NH)p, GTP-gamma-S, sodium fluoride, and forskolin, at a concentration of 0.1 mM, increased the basal activity of enzyme by 3.1, 5.7, 4.7, 3.6, and 7.8-fold, respectively. The stimulation of adenylyl cyclase by VIP was time, temperature, and membrane concentration dependent. Half-maximal enzyme activation (ED50) was very similar in rat and mouse peritoneal macrophage membranes (1.5 +/- 0.1 nM and 1.0 +/- 0.1 nM, respectively). However, VIP showed more efficacy in mouse macrophages membranes (about 3.1-fold basal values) than that in rat macrophage membranes (about 2.5-fold basal values). The relative potency of several peptides upon stimulation of adenylyl cyclase activity showed the following potency in both species: VIP = PACAP38 = PACAP27 > helodermin > PHI > secretin. On the other hand, a M(r)-45 kDa alpha s subunit of Gs protein was demonstrated by both ADP ribosylation and immunoblot in mouse and rat peritoneal macrophage membranes. The present results, together other previous, strongly suggest that VIP play an important role in the regulation of macrophage function. PMID- 8703999 TI - Chemically dependent physicians and informed consent disclosure. AB - Developments in law, professional guidelines, and public attitudes support informed consent disclosure by physicians who have been treated for chemical dependency. This view is built on the apparent materiality of the risk of relapse to informed treatment decisions by patients. Several considerations undercut this position. The probability is remote that a patient will be injured by a recovering physician who suffers an undetected relapse while being properly monitored. Monitoring by impaired physicians programs provides a more sensitive and specific mechanism for detecting relapsed physicians. Moreover, compromise of the privacy and employment rights of recovering physicians by consent disclosure is not justified if programs provide rigorous monitoring that protects the welfare of patients. Finally, required consent disclosure will reduce referrals of chemically dependent physicians to impaired physicians programs, thereby increasing the danger to patients. Limiting demands for required consent disclosure necessitates effective operation of impaired physicians programs. PMID- 8703998 TI - Learned helplessness and cocaine dependence: an investigation. AB - Low personal control and a sense of meaninglessness of life, attributes associated with a pattern of human learned helplessness, have also been described as contributing to the onset of adolescent drug use, as well as the maintenance of chronic substance abuse. However, despite its intuitive appeal as an etiologic factor, the absence of psychometrically sound and easily administered measures of learned helplessness has limited the ability of researchers to empirically test its role in the addiction process. Accordingly, the publication by Quinless and Nelson (1988) of a relatively brief Learned Helplessness Scale (LHS) led us to administer it to a sample of 30 consecutive cocaine dependent individuals seeking treatment for the first time at our facility. Our results indicated that the LHS was internally consistent when administered to this clinical sample and that theoretically meaningful and statistically significant relationships with other measures of psychological functioning were observed. Moreover, scores on the LHS were related to treatment retention/outcome. PMID- 8704000 TI - Comparison of DSM-III-R symptoms for alcohol dependence between patient self report and clinician interview or the Structured Clinical Interview for DSM-III R. AB - This study sought to determine which Diagnostic and Statistical Manual of Mental Disorders, Third Edition, Revised (DSM-III-R), symptoms of alcohol dependence were most sensitive to under-reporting by 78 inpatients from alcohol treatment programs. We hypothesized that patients would be more reluctant to report social/behavioral symptoms (lost time, hazardous behavior or major role interference, and reduced activities) than psychological or physiological symptoms. Patient endorsement of symptoms on a self-administered diagnostic questionnaire was compared with parallel items on the Structured Clinical Interview for DSM-III-R (SCID) and clinician interview. Bias and Prevalence Adjusted Kappas for individual symptom agreement ranged from -.02 to .87. Subjects endorsed fewer symptoms per category than either the SCID or clinician interview. Using a Student-Newman-Kuels post-hoc analysis at the p < .05 level, we found that mean agreement for the social/behavioral category was significantly lower than mean agreement for the other categories. PMID- 8704001 TI - Spirituality and the treatment of the dually diagnosed: an investigation of patient and staff attitudes. AB - Spirituality is a neglected area of study and research in the treatment of addictions. The role of spirituality in the treatment of the dually diagnosed has received particularly scant attention. One hundred and one patients on an in patient dual-diagnosis unit, as well as the 31 members of the nursing staff who treat them were surveyed. Patients and staff were questioned about their spiritual beliefs and what was the role of spirituality in the patients' recovery from addiction. Staff were questioned about their own spirituality and what they think the patients' level of spirituality is. In addition the staff were asked what they think the patients' view of spiritually is. Results indicate that the patients and staff are equally spiritually oriented. The patients view spirituality as essential to their recovery and value spiritual programming in their treatment more than some concrete items. The nursing staff underestimated both the patients' level of spirituality and this importance placed on spiritual issues. The authors suggest that more attention should be given by staff to spirituality in the treatment of this population. PMID- 8704002 TI - Rapid measurement of plasma methadone in a clinical setting using florescence polarization immunoassay. AB - Currently clinicians using methadone for treatment of opioid dependency or organic pain lack a simple, real-time, analytical method useful for adjusting methadone dose. Therefore, we evaluated the feasibility of measuring plasma methadone concentrations using florescence polarization immunoassay (FPIA) in an outpatient clinical setting. Using this technology, patient plasma methadone concentrations can be determined in approximately 20 minutes. The FPIA calibration curve was found to be linear over the methadone concentration range of 0 to 1600 ng/ml. Day-to-day run coefficient of variation was 5-10%, the within run coefficient of variation was 1.3-6.3%; the limit of quantification for the assay was 25 ng/ml. Calibration plots of HPLC and GCMS (mass fragment 72) plasma methadone control samples versus FPIA were also linear. A plot of HPLC plasma methadone versus FPIA for patient samples was identical to results for control samples (concentration range 0-1200 ng/ml). No significant amounts of the EDDP methadone metabolite were found in plasma. Based upon these findings, FPIA is clinically useful for monitoring plasma methadone concentration in outpatient settings. PMID- 8704003 TI - Urine toxicology as an outcome measure in drug abuse clinical trials: must every sample be analyzed? AB - Clinical trials designed to establish the effectiveness of a pharmacotherapy for the treatment of drug abuse typically call for the collection and analysis of three urine samples per week to detect changes in drug use patterns. Examination of over 16,500 urine samples collected from 225 subjects during a one year buprenorphine/methadone clinical trial indicates that analysis of one weekly urine sample from those collected on a three-times-per-week fixed schedule provides essentially the same outcome information as analysis of all three weekly urines. Further, the percent of opiate-positive samples is constant across weekday, indicating that a single urine, randomly selected from those collected each week, is a valid indicator of treatment performance. PMID- 8704004 TI - Sexual health issues in inflammatory bowel disease. AB - Alterations in sexual health caused by inflammatory bowel disease (IBD) may affect quality of life and disease status. IBD usually strikes adolescents or young adults, who are also facing developmental milestones important to sexual health. Issues include growth and development, body image, intimacy and sexual functioning, fertility, and pregnancy. A review of published research regarding these issues, in addition to suggestions for nursing assessment and interventions, is included in this article. Nurses must offer sensitive support and suggestions for coping. Nurses must be aware of the issues influencing sexual health when providing total care to clients with IBD. PMID- 8704005 TI - Trauma-induced severe acute pancreatitis in adults: conceptual review of pathophysiological events. AB - Trauma-induced severe acute pancreatitis in adults is used to demonstrate the complexity of a comprehensive conceptual pathophysiological review. The exemplar depicts a progression of pathological events evolving from pancreatic cell damage, as influenced by the stress response. Discussion of sequelae of the cardiovascular, respiratory, and renal systems is included, highlighting core concepts of inflammation, immunosuppression, fever, and impaired wound healing. Aspects of Neuman's Systems Model are introduced to guide the discussion. It is proposed that facilitating an in-depth conceptual understanding of pathology will enable nurses to improve "pattern" recognition, thereby enabling them to make quicker and more accurate assessments and diagnoses leading to treatment. PMID- 8704006 TI - A review of the transmission of viral hepatitis for gastroenterology nurses. AB - This article focuses on viral hepatitis because of the potentially devastating consequences of this disease group to gastroenterology nurses. Recent landmark work has reaffirmed the high risk of occupational exposure to viral hepatitis and the need for inoculation of all healthcare workers. Gastroenterology nurses, by virtue of their professional activities, are at particular risk. The various forms of viral hepatitis are reviewed and appropriate nursing interventions are discussed in this article. PMID- 8704007 TI - Withdrawal of nutritional support: a family's choice. AB - Facilitating the death of a loved one through the withdrawal of nutritional support is a difficult choice. This case study illustrates the ethical principles used by a family and a health team during the decision to terminate tube feeding and hydration for a patient. Patient autonomy, beneficence, justice, and professional integrity are discussed. Also addressed are use of ordinary versus extraordinary means of treatment, futility of treatment, and quality of life. Nursing interventions used to assist the family with their fears and concerns during the decision-making process, and eventually the dying process, of their husband and father are described. PMID- 8704009 TI - The first 1,000 SIGs. PMID- 8704008 TI - Reducing the cost of enteral feeding. PMID- 8704010 TI - Creon (pancreatin). PMID- 8704011 TI - The use of expandable metallic stents in patients with esophageal cancer: a case report. AB - Esophageal cancer has provided today's gastroenterology nurse with an opportunity to assist with an effective palliative treatment offering patients a better quality of life. This procedure is relatively new and requires a minimal hospital stay, thereby demanding that staff members quickly formulate and implement a personalized patient care plan. The case presented in this article details the first esophageal implant performed in New York State and the fifth in the United States. Although the basic care is similar to that of any patient undergoing a panendoscopy, this patient requires more emotional support and in-depth patient teaching concerning the palliative nature of the procedure and the postprocedural retrosternal pain to be expected. Objectivity is essential in the care of these patients because of their familiarity with staff members as a result of frequent visits to the Endoscopy Suite for other palliative treatments such as esophageal dilatations and laser treatments. PMID- 8704012 TI - [The value of life. Development of an assessment system for therapeutic strategies]. AB - The wide therapeutic supply which a present is available for diverse disorders, together with a excess of indiscriminate scientific information, occasionally complicates the possibility of acquiring a wide experience in the treatment of certain patients (limited experience). To acquire genuinely useful experience, it is necessary to accumulate simiular experiences from our environment, with it being necessary to precisely define, in an objective manner, concepts which until now are difficult to quantify (risk of mortality or morbidity, survival, quality of life, etc.). The definition of these parameters, shall favor the creation of homogenous groups (isoseverity indices), which is the first step in the evaluation of more complex conceps, such as; quality of life, increase of life expectancy, quality adjusted life years (QALY's), and beneficial application, all of which validated in diverse hospital media. Based on the previous, we would obtain other calculated parameters; the benefit (of the patient), the use (of the procedure), and the efficacy (of the physician or the unit). Once these parameters have been standarized, it shall be possible to compare them with the results published in other media, considered to be standards, with the aim of establishing the corresponding self-evaluation. The evaluation of the cost of each process, is a compulsory procedure, to produce, if possible, based on the previous results, a better distribution of the means without a decrease in the quality of assistance. With the aim of evaluating the results in an objective and quantifiable manner, we have worked these last years on developing assessment applied to the evaluation of predictive and isoseverity indices, such as the cost/benefit evaluation in critical, geriatric, and oncological patients, and in those with artificial nutrition in the home. The fruit of this labor has come together in the production of a computerized information system (SAETA), which builds de previously validated modules into is structure, thus permiting the storage and subsequent multifactoral analysis of the results. PMID- 8704013 TI - [Carbohydrates--no glucose--in parenteral nutrition. A fading concept?]. AB - An update is made on the binominal use/benefit of non-glucose carbohydrates (polyalcohols or polyols) in parenteral nutrition. The most recently published results, despite the controversy from different areas, have motivated the updating of the criteria metabolic aspects, indications and contraindications of the different carbohydrates alternatives of glucose (xylitol, sorbitol, glycerol, fructose), in the parenteral nutrition of patients with different degrees of aggression and situations of glucose intolerance or insulin resistance. It is known that, due to causes which are not entirely understood, non-glucose carbohydrates have not yet been considered as effective energy sources alternative to glucose in clinical situations described as unfavorable for their use. However, we may consider that in view of our own experience and that of previous publications, these carbohydrates have not only not decreased their use expectancy but, on the contrary, have increased as their advantages were clinically confirmed, without the appearance of major complications in this therapeutic modality. For this reason we consider that the understanding of the characteristics and the different metabolic aspects (pharmacological dose) of the polyols must make us review their clinical use, both alone as in different combinations. PMID- 8704014 TI - [A catalogue of fatty acids]. AB - Fatty acids structure and function is an area of renewed interest because of its effects on plasma lipids, biosynthesis of prostaglandins, leucotrienes and thromboxanes, and the obligatory demands of some fatty acids, especially for the newborn. Fatty acids are identified in three different ways: by the classical nomenclature, by its trivial name, and by the new methods also known as the omega system. These three different methods have created some confusion. The aim of this article is to revise fatty acids chemical structure and to compile a list of nutritional important fatty acids with the three different terminologies. PMID- 8704015 TI - [Hepatic metabolism of cholesterol]. AB - Cholesterol is an essential component of all tissues, as it is a part of the structure of cell membranes, and it is an immediate precursor of a series of essential substances such as vitamins, steroid hormones, and bile acids. Under physiologic conditions, the intake and output of cholesterol in the organism is coordinated and balanced with the aim of guaranteeing the availability of adequate amounts of cholesterol to satisfy the needs of the different tissues (fig. 1). Under pathological conditions there is an imbalance between these mechanism, which leads to an increase in the circulating levels of cholesterol, leading to pathological processes such as hyperlipemias, atherosclerosis and bile stones. The liver plays a central role in the regulation of the homeostasis of cholesterol. The molecule enters the liver in the form of chylomicrons and low density lipoproteins (LDL), through lipoprotein receptors, and this is also the most important organ for the de novo biosynthesis of cholesterol from acetyl coenzyme A, by means of a cascade enzyme reaction in which the enzyme 3-hydroxy-3 methyl glutaryl CoA reductase (HMG-CoA) is the key of the entire process. Cholesterol is found in the liver in the form of cholesterol esters or as free cholesterol. The two most effective ways of eliminating body cholesterol are found in the liver, with the degradation of the compound to bile acids and the biliary secretion of cholesterol. The conversion to bile acids takes place through a series of enzymatic steps in which the formation of 7-alpha hydroxycholesterol by the enzyme cholesterol 7-alpha-hydroxylase is the key of the process. The biliary secretion of cholesterol is 600 mg/day. Both the abundance and the universality of cholesterol in living things as its clinical implications emphasize the importance and interest of this compound. PMID- 8704016 TI - Some characteristics of nutrition metabolism and hormone control during the first 24 hours in neonatal calves. AB - Six male Holstein calves of spontaneous delivery and with jugular vein catheter were chosen to study the characteristics of nutrition metabolism and hormone control during the first 24 h of age after birth. The results showed that plasma fructose (Fru) and lactic acid (LA) concentrations were highest at birth, fell significantly during the first 2 h of life, and then dropped further until 24 h. Glucose (Glu) increased at 0.5 h after birth, followed by a gradual decrease until 8 h, with a general upward trend thereafter. Free fatty acid (FFA) was very low at birth, rose rapidly within 2 h of live, and kept a relatively high concentration thereafter. Total protein (TP) had not obvious change and fluctuated at a low level during the first 24 h of age. The rise of insulin (Ins) at 1 h of age was observed, with a downward tendency thereafter. Plasma cortisol (Cort) level was high during 1 h of age and tend to drop thereafter. An obviously rising tendency of T4 and T3 was found during the first 4 h of life, followed by their keeping relatively high levels although decreasing constantly in neonatal calves. PMID- 8704017 TI - [Comparative study of two lipid emulsions in the parenteral nutrition of the septic patient]. AB - The nutritional support is essential in the treatment of the septic patient, but the optimal substrates, have not been well defined, although several studies advise the use of solutions with a high proportion of branched amino acids. The use of lipids in the Parenteral Nutrition (PN) of these patients is presently accepted by all authors, although there is no consensus of the composition of the mix to be used. We have done a prospective, randomized study in a group of critical septic patients who received PN. All cases were used a solution with a high proportion of branched amino acids and glucose. 40% of the non-protein calories were administered as lipids. Based on this, two groups were formed. Group 1 medium chain triglycerides and long chain triglycerides (MCT/LCT) in a ratio of 1:1, and Group II LCT. After ten days, both groups showed an increase in hepatic cholestasis enzymes and an improvement of the parameters of protein anabolism and catabolism. Group 1 showed a significantly higher increase in the retinol birdring protein and the improvement of the nitrogen balance. Besides this, the global analysis of all the parameters of protein anabolism and catabolism showed a more favorable evolution, with statistical significance, in those who received MCT/LCT. Therefore, the MCT/LCT in the PN of the septic patient who is given a solution rich in branched amino acids, achieves a greater protein saving and a faster recevesy of the markers of protein synthesis, than those with LCT. Both groups show an increase in cholestasis enzymes. PMID- 8704018 TI - [Influence of energy intake and body mass index in the incidence of cancer of the breast. Case-control study in a sample from 3 Spanish hospital populations]. AB - A case-control study has been designed to analyze the possible role of the energy intake and obesity, judged by the body mass index, on the incidence of breast cancer in a group of 275 women (139 cases and 136 controls), coming from three hospital populations (Madrid, Merida, and Santiago de Compostela). There are no significant differences in the of cases and controls, which, additionally, does not seem to be significantly related to the risk of breast cancer. On the other hand, the weight and the body mass index of women with breast cancer is significantly higher than that of the controls. The oods ratio, as a measure of the relative risk for breast cancer, shows that a high body mass index behaves as a risk factor (OR = 2.49; 95% IC, = 1.33-4.67, Chi2 = 8.30), a trend which remains after adjusting for energy. When considering the pre or post menopausal status, it is observed that a body mass index higher than 29, adjusted for energy intake, behaves as a risk factor only in the post-menopausal women (OR = 2.28; 95% IC = 0.95-5.46 Chi2 = 3.41). PMID- 8704019 TI - [Influence of resting the digestive duct in gastric emptying. Comparison of 2 nutritional modalities: glucidic enteral and total parenteral nutrition]. AB - Gastrointestinal motility is altered in several clinical situations in which Total Parenteral Nutrition (TPN) is indicated. Nevertheless, the effects of TPN on motility are not well known. In this study, Wistar rats (250-280 g) were divided into two groups were defined according to the route of administration of the nutrients: Group A (with TPN: 100% of the total non-protein calories administered as glucose), and Group B (solid enteral nutrition, but with the infusion of normal saline under the same conditions as in the previous group). Each group was subjected to three different infusion times: 24 hours, 3 days, and 5 days post-intervention, after which the gastric emptying was measured by means of the phenol red test. TPN delayed emptying in all the times with respect to group B, although significant differences were only found after 3 days of infusion. PMID- 8704020 TI - [Incidence of phlebitis in patients with intestinal inflammatory disease undergoing peripheral parenteral nutrition]. AB - The aim of this study is to evaluate the incidence of phlebitis in patients with inflammatory bowel disease under parenteral nutrition treatment (700 mOsm/ml) via peripheral vein. Results will be used to develop an standar protocol. 30 patients were sampled and the cannulation was performed by nursery personnel. Daily, incidence of phlebitis was evaluated with a gradual scale ranging in 5 possible degrees from absence of signs to thrombophlebitis. 86.6% of the patients received the feeding through a Teflon catheter with a fine bore of 20 gauge. Cannulas were mostly situated in the basilic vein. Development of phlebitic signs was proportional to the duration of infusion. When phlebitic signs appeared, via of cannulation was not changed in 33% of those patients. PMID- 8704021 TI - [Lipid vehicles for the parenteral administration of drugs (1): emulsions]. AB - To increase the efficiency of medication used in therapy and diagnostics, there have been, during the last decades, important advances in the inclusion techniques hereof, in vehicles which give greater selectivity and tolerance. Among the colloidal vehicle systems of medications with different medications, the object of the present review study. Therefore, we analyze the rationale of their use as a vehicle of medication, their physical and chemical characteristics, the ways of elimination after their administration, as well as the formulations in oil-water emulsions for the parenteral administration of diferent active substances such as diazepam, propofol, methohexital, ethomidate, amphotericin B, cytostatics, and cyclosporine, among others. The results of published studies on the efficiency and therapeutic applications of these are presented. PMID- 8704022 TI - ICN calls for more protection against HIV/AIDS. PMID- 8704023 TI - Making decisions about circumcision. PMID- 8704024 TI - Health care reform. Regionalization update. PMID- 8704025 TI - Health care reform. Why nurses should care. PMID- 8704026 TI - Health care reform. Mission nurses get involved. PMID- 8704027 TI - Continuing competence: a shared responsibility. PMID- 8704028 TI - Standards in action: taking charge of nursing practice. PMID- 8704029 TI - Distribution of prolyl oligopeptidase in human peripheral tissues and body fluids. AB - Prolyl oligopeptidase (EC 3.4.21.26) activity was measured in human tissue homogenates and body fluids. The enzyme was ubiquitously present, revealing high activity in renal cortex, epithelial cells, fibroblasts, testis, lymphocytes and thrombocytes. The activity in the body fluids was low. Prolyl oligopeptidase activity was significant higher in tumours of prostate, lung and sigmoid, than in the healthy tissues. Sera of individuals suffering from HIV infection, malaria, prostate cancer or benign prostate hypertrophy contained lowered activity. Interestingly, the low serum activity during prostate carcinoma increased upon medical treatment with anti-androgens. This suggests hormonal control of the gene transcript. A positive correlation with angiotensin converting enzyme activity in hypertensive patients was demonstrated and this further supports the possible involvement of prolyl oligopeptidase in the renin-angiotensin system and in the pathogenesis of hypertension. PMID- 8704030 TI - Analysis of the in vitro secretory activity of human pituitary adenomas: modification of corticotropin release from adenoma tissue explant cultures by addition of a human plasma ultrafiltrate bioactive fraction. AB - The lack of control of tumour behaviour is manifested in different ways, depending primarily on the type of tumour. This results in numerous problems of tumour diagnosis and therapy. In the case of "benign" tumours, like pituitary adenomas, in vitro studies are often used for evaluation of the tumour. The use of tissue explant cultures of human pituitary adenomas and the comparison of the feature of cultured tumours with their behaviour in vivo showed that corticotropin is released not only from the tumours associated with Cushing's disease, but also from clinically non-functioning tumours. Hence, it was supposed that the release of corticotropin in vivo from non-secreting tumours is probably under the influence of certain neuroendocrine and/or systemic humoral factors. To test this possibility, samples of 22 tumours were cultured in plain culture medium or in the presence of the "human plasma ultrafiltrate bioactive fraction" (tentatively termed as TBP) prepared by anion-exchange chromatography. In the presence of TBP the release of corticotropin was strongly inhibited in adenomas showing relatively high spontaneous secreting activity in vitro (> 200 ng/l in 24 hours), while immunohistochemistry of these tumours indicated accumulation of corticotropin inside the cells. In contrast, TBP stimulated corticotropin release from tumours that showed relatively low basic corticotropin release (< 200 ng/l in 24 hours), with no obvious change in cellular corticotropin immunoreactivity. Such a dual activity of TBP was not observed for 8 samples of adenomas cultured in the presence of surrounding pituitary tissue, probably because TBP did not affect corticotropin secretion by the normal pituitary cells (as indicated by immunohistochemistry). From these results, it appears that TBP could be one of the humoral factors involved in the regulation of corticotropin release from pituitary adenoma tissue. Its possible involvement in the regulation of corticotropin release from normal pituitary tissue, however, is uncertain. PMID- 8704032 TI - Serum amyloid A protein in the prediction of postburn complications and fatal outcome in patients with severe burns. AB - Fifty adults hospitalized with extensive burns formed the basis of the present study. Serum amyloid A protein, C-reactive protein, alpha 1-antichymotrypsin and alpha 1-acid glycoprotein were measured in serum samples taken on admission, and 3 and 7 days later. Fatal outcome was observed in 13 out of 14 (93%) patients with serum amyloid A protein over 100 mg/l on admission and in only 2 of the remaining 36 (6%) patients with serum amyloid A protein below 100 mg/l. The median serum amyloid A protein concentration on admission in 15 patients with fatal outcome was 134 mg/l, and only 30 mg/l in 35 patients who recovered (p < 0.00005). As a reference value, the level of 100 mg serum amyloid A protein per litre on admission gave an evident predictive value (93%) and sensitivity (87%) for fatal outcome. The difference between serum amyloid A protein concentrations in patients with complications (median 642 mg/l) and those without complications (median 250 mg/l) was statistically very significant (p = 0.0003) three days after burn injury. The level of 400 mg/l as a reference value 3 days after burn injury gave a reasonable predictive value (80%) and sensitivity (74%) for the development of postburn complications, but patients who died did not develop a hypermetabolic reaction and their serum amyloid A protein concentration remained below 400 mg/l, despite high serum amyloid A protein concentrations observed on admission (above 100 mg/l). No statistical significance was observed for the other 3 acute phase proteins investigated in this study. PMID- 8704031 TI - In vivo effects of 13-cis retinoic acid treatment on the concentration of proteins and lipids in serum. AB - A number of serum components, whose concentrations or gene expression have been shown to be modulated by all-trans retinoic acid in vitro, were monitored in patients before and during treatment with Roaccutane (13-cis retinoic acid, 40-60 mg/day) for severe acne. The 13-cis retinoic acid concentration in serum rose from 5.25 +/- 1.09 to 593 +/- 65 nmol/l (mean +/- SD) 24 h after the latest dose. The concentration of all-trans retinoic acid in serum under Roaccutane treatment was measured in model experiments and shown to be 10-20 nmol/l i.e., 2-4 times the basal levels (4.65 +/- 0.85 nmol/l) when the 13-cis retinoic acid concentration was 370-980 nmol/l. The concentrations of creatine kinase-MB, apolipoprotein B, total cholesterol and LDL cholesterol increased significantly while the other measured serum components, including lipoprotein lipase activity, were unaffected by Roaccutane treatment. PMID- 8704033 TI - Inter-method differences in the measurement of some specific plasma proteins: commutability of control materials. AB - We compared the inter-method differences shown by control materials and by patients' sera for the measurement of some plasma proteins in the same pair of analytical systems. Sets of 100 to 110 samples of patients' sera and of 18-19 control materials, including the recently available CRM 470, were assayed with up to five automatic analytical systems, in two different experiments. About 5500 values were produced and assessed statistically. Materials (either patients' sera or control materials) were considered non-commutable (i.e. exhibiting significantly different inter-method behaviour) when their distance from the regression line in a stated pair of methods exceeded 3 standard deviations. According to this criterion, less than 1.5% of the patients' sera, and an even lower proportion of control materials were non-commutable. However, the inter method behaviour of control materials was usually slightly different from that of patients' sera. Some systematic inter-method difference in the measurements on patients' sera may therefore exist, even though inter-method equivalence has been demonstrated with control materials. PMID- 8704034 TI - Thyroglobulin determined in thyroid fine needle aspiration biopsies by radial immunodiffusion and electroimmunodiffusion. AB - Radial immunodiffusion and electroimmunodiffusion were used to measure thyroglobulin, the main component of thyroid colloid, in thyroid fine needle aspiration biopsies. A linear relationship was established between precipitation ring diameter and thyroglobulin concentration by radial immunodiffusion (0.5-3.0 g/l), and between "rocket" height and thyroglobulin concentration by electroimmunodiffusion (0.1-2.0 g/l). A nearly complete correlation was observed between the two methods (r = 0.97). In radial immunodiffusion the ring diameter is dependent on time of diffusion and on the antiserum concentration in the agar gel. In this study, the observation time was standardised at 48 h, and the rabbit anti-thyroglobulin serum concentration at 26 ml/l. The intrathyroidal concentration of thyroglobulin was determined by radial immunodiffusion and the thyroid find needle aspiration biopsy of 45 thyroid tumours with different cytological-laboratory- and clinical diagnoses. It was found that in colloid nodules or cysts thyroglobulin is markedly higher than in euthyroid nodular goitre (13.7 +/- 11.9 g/l vs. 1.35 +/- 0.8 g/l, p = 0.005). In conclusion radial immunodiffusion and electroimmunodiffusion are precise, easy to perform, low cost, non polluting methods, which do not require high sample dilution (in contrast, high sample dilution is necessary for measurement of thyroglobulin in thyroid fine needle aspiration biopsy by radial immunodiffusion). Measurement of thyroglobulin in thyroid fine needle aspiration biopsy provides a quantitative estimate of colloid, an important marker in the differential diagnosis of thyroid nodules. PMID- 8704035 TI - Further development of flow-through electrophoresis. AB - Recent improvements in the field of flow-through-electrophoresis are presented. These improvements make it possible to run flow through electrophoresis with the same precision, but more economically and more easily than the formerly described apparatus. This was achieved by limiting the original system to slab gels only. This leads to a considerable reduction in the size of the apparatus and to simplification of its handling. Although the gel width is reduced to 10 mm, the apparatus is suitable for the analysis of mg as well as microgram quantities. The usefulness of this method is demonstrated by the analysis of samples obtained during the isolation and purification of collagen type II, and by the analysis of insulin samples and other substances. The procedure is also suitable for SDS electrophoresis under flow-through conditions, enabling a simple and fast separation of proteins and determination of their molecular masses. PMID- 8704036 TI - Preanalytical procedures in the measurement of ionized calcium in serum and plasma. AB - To investigate the effects of sampling routines, ionized calcium (corrected for pH) was measured after sampling in different types of tubes and under fasting and postprandial conditions. The results are discussed with respect to analytical goals. Serum samples, collected in vacuum tubes and spun after coagulation, could be safely measured during 24 h. This time window was expanded to 48 hours by using gel-containing tubes. Properly prepared specimens withstand mail transportation. Different brands of gel-containing tubes gave different results, and these differences were clinically significant. An "American-type" breakfast causes an average decrease of corrected ionized calcium ion concentration of 0.04 mmol/l and an increased interindividual variation of results. Characterisation of reference intervals should include the "nutritional status" and the brand of vacuum tube used. PMID- 8704037 TI - Analysis of breath alcohol via infrared spectrophotometry: predicting false ethanol results by application of the base-line method to vapor phase infrared spectra. AB - Toluene and a mixture of the isomeric xylenes are common organic solvents that have been implicated in false ethanol results produced by older models of the Intoxilyzer 5000, a breath alcohol analyzer that uses infrared spectrophotometry to quantitate ethanol in breath samples. A straightforward method is described which predicts this type of interference in analyses conducted on such models. The methodology relies on a comparison of the vapor phase infrared spectra of toluene and xylene with the corresponding spectrum of ethanol, using the base line method to estimate relative absorbances at the two analytical wavelengths employed by the Intoxilyzer 5000. The point is also made that, given the potential for such interference, a model of the Intoxilyzer 5000 has been developed that has the capability to compensate for this problem. PMID- 8704038 TI - Negative methaemoglobin results obtained with the IL 482 CO-Oximeter. PMID- 8704039 TI - False positive immunoassay results for cholecystokinin with a phosphate buffer containing bovine serum albumin. PMID- 8704040 TI - Evaluation of a reference material for glycated haemoglobin. AB - The use of lyophilized blood as a reference material for glycated haemoglobin was investigated with respect to IFCC criteria for calibrators and control materials. Ninety-two laboratories, using 11 methods, detected no changes in glycated haemoglobin content when the lyophilizate was stored for one year at 4 degrees C. Affinity chromatography, HPLC, electrophoresis and immunoassay detected no changes following 18 months storage at -84 and -20 degrees C. Samples for HPLC are stable at 4 degrees C for one year, and 5 years at -20 degrees C. For the other three methods, samples are stable for 5 years at 4 degrees C. At 4 degrees C, reconstituted samples are stable for 2 days (HPLC) and 7 days (other three). Lyophilization does not cause matrix effects and inhomogeneity, since mean glycated haemoglobin and reproducibility for lyophilized samples and whole blood were similar. The coefficient of variation for vial filling precision was 0.59%. We conclude that lyophilized blood samples can be used as calibrators and control materials. Their use as calibrators, following assignment of the HbA(1c) value by HPLC, may contribute, in the interim, to the standardized interpretation of long term diabetic control. PMID- 8704041 TI - Evaluation of a second-generation thyrotropin automated immunoassay. AB - We evaluated the analytical performance of the Vista automated immunoassay system for human thyrotropin determination. The operating characteristics as well as the analytical performance were assessed. The Vista human thyrotropin immunoassay showed a minimal detection limit of 0.08 mU/l and a functional sensitivity of 0.12 mU/l. The system meets the criteria for second-generation human thyrotropin assays. PMID- 8704042 TI - CA 15-3: a multicentre evaluation of automated and manual tests. AB - The introduction of a new and automated CA 15-3 immunoassay (IMx Abbott) prompted us to compare the analytical performance of this new test with established tests from CIS ELSA, Sorin, and Boehringer Mannheim in a multicentre study. CA 15-3 measurements in blood samples of breast tumour patients, comparison of intra- and inter-assay variation, dilution linearity, and lower limit of detection are described. The study showed improved precision for the automated over the manual test systems (intra-assay variation: IMx < 5%, CIS ELSA 4-9%, ES 300 < 3% and ETI Sorin > 10%; inter-assay variation: IMx < or = 8%, CIS ELSA < or = 19%, ES 300 < or = 9% and ETI Sorin < or = 27%). Results on patients' samples (n = 101 to 184) showed highly comparable results; IMx vs CIS ELSA site 1: r = 0.950; IMx vs CIS ELSA site 2: r = 0.998; IMx vs ES 300: r = 0.980; IMx vs ETI Sorin: r = 0.931. Slopes of regression lines varied from 0.666 for IMx vs ETI Sorin to 0.988 for IMx vs CIS ELSA (site 1, where heparin plasma was used instead of serum as recommended by the manufacturer found to be only slightly dependent on assay ranges analysed by statistical procedures applied. Despite good correlations between methods, it is recommended that samples collected in the follow-up of disease and at higher CA 15-3 concentrations are analysed by the same test; a changeover to another test is not encouraged. PMID- 8704043 TI - Enzymatic determination of phosphatidylcholine, sphingomyelin and phosphatidylglycerol in lipid dispersions, blood cell membranes and rat pulmonary surfactant. AB - A new micromethod for the determination of sphingomyelin in samples suspended in aqueous solutions, and modified micromethods for determining phosphatidylcholine and phosphatidylglycerol were used to determine phosphatidylcholine and sphingomyelin (detection limits of 1.8 mumol/l), and phosphatidylglycerol (detection limit of 2.3 mumol/l) in lipid dispersions, membranes from sheep erythrocytes and platelets, and pulmonary surfactants from rats of different ages and rats maintained under normobaric hyperoxia for 2 days prior to their sacrifice. The procedures are easy to perform, accurate, require less sample than conventional methods and can also be applied directly to aqueous samples. Phospholipase C and sphingomyelinase were used to release phosphorylcholine from phosphatidylglycerol and sphingomyelin, respectively. The choline released from phosphorylcholine by alkaline phosphatase is reconverted to phosphorylcholine by ATP and choline kinase. In the phophatidylglycerol determination, phospholipase D was used to release glycerol and phosphatidate. The glycerol formed was converted to glycerolphosphate using ATP and glycerol kinase. In all cases, the ADP thus formed was determined by following the enzymatic conversion of NADH to NAD at 340 nm in an coupled pyruvate kinase/lactate dehydrogenase system. Significant variations in the phospholipid composition of rat pulmonary surfactant were found during development; in particular there was an increase in the phosphatidylglycerol content of adult rats as compared with younger rats. Hyperoxia produced changes in the phosphatidylglycerol content of surfactant from adult rats, but not from 2-day old rats. PMID- 8704044 TI - Relationship between bone metabolism and plasma cytokine levels in patients at risk of post-transplantation bone disease after bone marrow transplantation. AB - Bone marrow transplant recipients may carry an increased risk of bone diseases, involving numerous factors that affect bone mineral metabolism. Interleukin-6 is a potent stimulator of bone resorption in vivo. The soluble fraction of interleukin-6 receptor is reported to trigger osteoclast formation by interleukin 6 in vitro. In a cross-sectional study we measured serum bone alkaline phosphatase concentrations and the urinary excretion of pyridinium cross-links in 21 patients after bone marrow transplantation, and investigated the relationship between these values and those for the plasma levels of interleukin-6 and soluble interleukin-6 receptor. Following bone marrow transplantation female--but not male--patients showed higher serum bone alkaline phosphatase values than age- and sex-adjusted controls (p < 0.05). Both female and male patients were characterized by increased urinary excretion values of pyridinium cross-links (p < 0.05). In contrast to a marked increase of interleukin-6 plasma levels (p < 0.001) no significant difference in the soluble interleukin-6 receptor levels was found between patients and apparently healthy persons (p = 0.838). Multiple regression analysis (taking into account different variables of the immunosuppressive regimen applied) revealed the plasma concentration of interleukin-6 as an independent predictor of the urinary excretion of pyridinium cross-links (p < 0.05). In conclusion, in patients following bone marrow transplantation, these findings indicate (a) an increase of bone formation in female--but not in male--patients possibly reflecting primary ovarian failure and (b) an enhancement of bone resorption possibly mediated by circulating interleukin-6. PMID- 8704045 TI - Association between deletion polymorphism of the angiotensin-converting enzyme gene and cerebral atherosclerosis. AB - We investigated deletion polymorphism in the gene for angiotensin-converting enzyme in patients with angiographically verified cerebral atherosclerosis. Genotypes were determined by the polymerase chain reaction with oligonucleotide flanking of the polymorphic region of intron 16 of the angiotensin-converting enzyme gene. Results of angiotensin-converting enzyme genotyping showed 46% of 50 studied patients to be homozygous for the DD allele, whose prevalence was significantly increased as compared with a group of controls without atherosclerotic changes. In this control group, the following genotypes were observed (%): II = 24, ID = 52 and DD = 24. The frequency of the I and D alleles in the group of patients with cerebral atherosclerosis was 0.28 and 0.72, respectively, whereas in the group without atherosclerosis it was 0.50 for both. Furthermore, in the present study, the DD genotype was associated with a high level of serum angiotensin-converting enzyme activity, total and LDL-cholesterol and triacylglycerol. A newly established association between DD genotype and cerebral atherosclerosis, detected even in our small group, supports the view that angiotensin-converting enzyme polymorphism might be indicative of the development of cerebral atherosclerosis. PMID- 8704046 TI - Glutamate-induced efflux of protein, neuron-specific enolase and lactate dehydrogenase from a mesencephalic cell culture. AB - A mixed mesencephalic cell culture damaged by glutamate was used as a model to study the efflux of lactate dehydrogenase and neuron-specific enolase from neuronal cells into the culture medium. Glutamate toxicity was induced in sister cultures by 15 min exposure to 100 mumol/l glutamate in a Ca2+ containing salt solution. Cell injury was monitored 24 h later by measuring the lactate dehydrogenase activity and the neuron-specific enolase content in the cells and in the culture medium. The neuronal cell damage is reflected by an efflux of neuron-specific enolase and lactate dehydrogenase from the cells and an increase of lactate dehydrogenase catalytic activity concentration and neuron-specific enolase mass concentration in the culture medium. It was found that the efflux fraction calculated from estimations of the cells was clearly higher than the efflux fraction calculated from estimations of the amount of enzymes found in the culture medium. Calculations of the recovery of lactate dehydrogenase and neuron specific enolase and experiments designed to study the efflux of lactate dehydrogenase and neuron-specific enolase during incubation and washing showed that higher amounts of neuron-specific enolase are released than lactate dehydrogenase. A close correlation was found between the glutamate-induced changes of the neuron-specific enolase efflux fraction, based on enzyme determinations of the cells, and the change of the microscopically counted neuron specific enolase immunoreactive cell numbers. This indicates that the determination of the neuron-specific enolase efflux fraction (cells) is an accurate and sensitive marker of damaged neurons. The lactate dehydrogenase efflux fraction seems to be less sensitive for the quantitation of neuronal cell damage; in addition, it depends not only on the neuronal damage but also on the proportion of neurons in the cell culture. PMID- 8704047 TI - Indirect examination of exocrine pancreatic function in vivo by determination of chymotrypsin activity in animal intestine using a synthetic substrate. AB - Using the substrate N-acetyl-L-tyrosyl-p-aminobenzoic acid, we determined chymotrypsin activity in the small intestine of calf, pig, and poultry. Orally administered N-acetyl-L-tyrosyl-p-aminobenzoic acid is enzymatically cleaved in vivo, and the released p-aminobenzoic acid is determined by HPLC. We found that the p-aminobenzoic acid concentration in plasma and urine was significantly influenced by the feeding of soya flour. After soybean flour feeding, the p aminobenzoic acid concentration significantly increased in the plasma of calves and hens, in contrast to pigs, where the p-aminobenzoic acid concentration significantly decreased. This shows that the oral administration of N-acetyl-L tyrosyl-p-aminobenzoic acid with subsequent determination of p-aminobenzoic acid is suitable for the estimation of exocrine pancreatic function and for determination of changes in intestinal proteolytic activity caused by antinutritive substances. PMID- 8704048 TI - Antibodies to Tamm-Horsfall protein subunits prepared in vitro, in patients with acute pyelonephritis. AB - The aim of this study was to determine antibodies to Tamm-Horsfall protein subunits in patients with acute pyelonephritis. The protein subunits used in this determination were prepared by chemical treatment of Tamm-Horsfall protein isolated from the urine of healthy individuals. Values for IgG and IgA were significantly higher (p < 0.05 and p < 0.01 respectively) in patients than in healthy persons, while IgM class antibodies were significantly higher only in the test performed with subunits obtained with 8.3 mol/l acetic acid (THP-A) (p < 0.05). Values for all three classes determined in the test with THP-A were significantly higher in patients with vesicoureteral reflux than in patients with normal radiological findings (p < 0.05). Antibodies to Tamm-Horsfall protein subunits isolated from the urine of patients with acute pyelonephritis should also be determined. PMID- 8704050 TI - Cholesterol concentrations in diabetic patients in Nnewi, Nigeria. AB - Twenty out-patient and twenty in-patient adult Nigerian type II diabetics and twenty healthy subjects matched for sex, age and weight, were studied at Nnamdi Azikiwe University Teaching Hospital, Nnewi, Nigeria. Mean total cholesterol and fasting blood glucose concentrations were higher in the diabetics. The out patients had significantly higher total cholesterol and fasting blood glucose values than their in-patient counterparts. The possible cardiovascular risk of the raised cholesterol level in the diabetics was discussed. We recommend that for effective management of diabetic patients, periodic measurement of cholesterol level is necessary. An integrated enlightenment programme to acquaint the out-patients with the health benefits of dietary and drug compliance and occasional hospitalisation for better monitoring would be desirable. PMID- 8704049 TI - Analytical specifications of reference methods compilation and critical discussion (from the members of the European EQA-Organizers Working Group B). AB - We present a compilation of published data for accuracy, precision, and measurement design for analytes that, currently, are of major interest for European reference laboratories. These data are compared with recent recommendations for performance of reference methods to be used within networks of European reference laboratories. In addition, we review the literature on reference methods and related topics. PMID- 8704051 TI - Binding of anti-double stranded (ds) DNA-positive sera to denatured (d) DNA and synthetic poly[dA-dT] x poly[dA-dT] double stranded copolymer in an ELISA format. AB - Using an ELISA assay anti-nuclear antibody-positive sera from 300 patients with various immune-related diseases and 64 anti-nuclear antibody-negative sera were analysed for binding to S1-nuclease-treated double stranded (ds) DNA. In addition, the pattern of reactivity of 50 selected anti-dsDNA-positive sera was established using denatured (d) DNA and poly[dA-dT] X poly[dA-dT] double-stranded alternating copolymer (dAT) as additional DNA antigens. None of the 64 anti nuclear antibody-negative sera and 76 of the 300 anti-nuclear antibody-positive sera (25%) were anti-dsDNA-positive. Of the anti-nuclear antibody-positive and anti-dsDNA-positive sera, 48 (63%) were from systemic lupus erythematosus patients, and 7 (9%) from rheumatoid arthritis patients, whereas 21 patients (27.6%) suffered from various immune and non-immune related diseases. Anti-dsDNA positive reactivity was highly correlated with dDNA and dAT reactivity (r = 0.906, p < 0.0001 and r = 0.93, p < 0.0001, respectively). Although the majority of the 50 selected (37 systemic lupus erythematosus and 13 non-systemic lupus erythematosus) anti-dsDNA-positive sera concomitantly bound to both additional antigens, 7 of these (14%) did not bind to dAT, and 2 (4%) did not bind to dDNA. Anti-dsDNA-positive sera (n = 37) showed a similar pattern, in which 8.1% and 2.7% of sera did not bind to dAT and to dDNA, respectively. In contrast, anti dsDNA-negative sera from various immune-related diseases bound either ssDNA (12.5%) or dDNA and dAT (12.5%). These data suggest that dsDNA and dAT-based assays detect similar but not identical specificities in the sera of patients suffering from systemic lupus erythematosus and in a proportion of non-systemic lupus erythematosus patients. PMID- 8704052 TI - A rapid ELISA test for detection of human paraproteins. AB - A rapid ELISA test for detection, characterization and quantification of human paraproteins was developed. The proposed method is a sandwich ELISA, where the capture antibody is specific for a given heavy chain (gamma, alpha or mu) and the labelled antibody is specific either for kappa or for lambda light chain. Both standard and patient sera are tested with all six possible antibody combinations. Each paraprotein produces a significant increase in titre (as compared with standard) only when tested with the relevant pair of antibodies. This enables the determination of the isotype and light chain type of the paraprotein and the evaluation of its relative quantity in patient serum. The accuracy of the assay (relative deviation) varies from 0.04 for gamma lambda to 0.19 for alpha kappa. The cut-off values for each type of polyclonal immunoglobulin were determined with 200 healthy donor sera. 103 patient sera were analysed. ELISA data are in good agreement with M-component and other clinical data. PMID- 8704053 TI - Possible significance of advanced glycation end products in serum in end-stage renal disease and in late complications of diabetes. AB - Advanced glycation end products were determined immunologically in blood from diabetics, patients with renal failure and subjects with various other diseases. Elevation of advanced glycation end products levels in serum is not confined to the diabetic state but was also observed in some severely ill subjects. Patients with end-stage renal disease on dialysis displayed high advanced glycation end products levels in serum, irrespective of the presence or absence of diabetes. In contrast, advanced glycation end products levels in uraemic subjects not treated by dialysis were not different from controls. In diabetics, levels of advanced glycation end products were related to the state of late complications. Diabetics without sequelae showed advanced glycation end products within the normal range, whereas in the presence of late complications mean advanced glycation end products levels were elevated. In the case of retinopathy the increase in advanced glycation end products was associated with the severity of retinal status. PMID- 8704054 TI - In vivo interference of heparin bolus injection with photometric continuous lipase determination. AB - The new photometric lipase determinations use diacylglycerols as substrates in a micellar solution, whereas the turbidimetric method is based on emulsified triacylglycerols degradation. Extrahepatic lipoprotein lipase, which is well known to be liberated by heparin from the capillar binding sites, may cleave triacylglycerols as well as diacylglycerols. When a heparin bolus injection is given to a patient, the released lipoprotein lipase renders high lipase activities in blood misleading for a pancreatic disorder. PMID- 8704055 TI - Evaluation of the fully automated Cobas Core enzyme immunoassay for the quantitation of antibodies against hepatitis B virus surface antigen. AB - The Roche Cobas Core automated enzyme immunoassay of antibodies against hepatitis B virus surface antigen (anti-HBs) was evaluated using a protocol complying with the Societe Francaise de Biologie Clinique guidelines. Results showed good precision (CV below 6 and 9% for intra- and inter-assay precision) and accuracy (according to the Paul-Ehrlich-Institute standard); the detection limit of the test was 2 IU/l. The manufacturer's specifications were confirmed and a satisfactory correlation (r = 0.901) was found with an automated method (IMx, Abbott) used for comparison. PMID- 8704056 TI - Dilution of citrated plasma. PMID- 8704057 TI - The performance of the knowledge-based system VALAB revisited: an evaluation after five years. AB - In 1988, inundated by the tedious work of validation of laboratory reports in a large hospital biochemistry laboratory, we designed VALAB, a knowledge-based system specially dedicated to this iterative function. Coping at first with a few biochemical tests, the program has been progressively expanded to forty-five common chemical tests. Simultaneously some new rules have been introduced to "weight" the conclusion in different circumstances and rules taking into consideration some clinical data have also been written. Moreover the program moved to other disciplines, pH and blood gases, haematology and coagulation. Accordingly the evaluation protocol has been modified, incorporating a new step, the consensus decision of the pathologists, operating within the initial protocol and based upon the various criteria of epidemiology. These major changes and improvements have led us to check and describe again the performance of this updated VALAB knowledge-based system. PMID- 8704058 TI - A multicentre evaluation of tumour marker determinations using the automatic Enzymun-Test Systems ES 300 and ES 600/700. AB - A multicentre evaluation of the determination of carcinoembryonic antigen (CEA), the cancer antigens CA 15-3, CA 19-9, CA 72-4 and CA 125 (II generation), the cytokeratin 19 marker Cyfra 21-1 and alpha-foetoprotein (AFP) using the Enzymun Test System (ES 300 and ES 600/700) was performed in 23 laboratories. The tumour markers were measured in a total of 4266 human serum samples. The intra-assay precision was less than 5% in 80% of all serum samples investigated and in 95% of the serum samples at or above the cut-off level of the tumour markers. Inter assay precision was less than 10% in 86% of the marker determinations. The interlaboratory survey also showed high reproducibility for the determination of all the tumour markers. In 3 laboratories the results of CA 15-3 in 283 serum samples were compared with the IRMA method of CIS bio international. The regression coefficient, r, was 0.967. In 4 laboratories the results of CEA in 312 samples were compared with the results obtained on the IMx analyser. The regression coefficient, r, was 0.967. In benign gynaecological diseases, CA 125 (II) was most frequently elevated in endometriosis. In gastrointestinal diseases it was proven that CEA is still the marker with the highest sensitivity as compared with CA 19-9 and CA 72-2 (59% with healthy controls as the reference group and 44% with patients having benign gastrointestinal disease as the control group). In pancreatic cancer CA 19-9 showed the highest sensitivity (78% and 62% respectively). In gastric cancer the three markers did not show statistically different results. When the gastric cancer patients were divided according to stage, CA 72-4 appeared to be more sensitive than CA 19-9 only in stage IV. PMID- 8704059 TI - Report on the symposium "Drug effects in Clinical Chemistry Methods". AB - The aim of the symposium was to establish a list of 20-30 drugs and to determine test concentrations (at therapeutic levels and above) that would indicate interference to clinical chemistry methods in serum and plasma. The following agents were chosen: Acetaminophen, Acetylcysteine, Acetylsalicylic acid, Ampicillin, Ascorbic acid, Ca-Dobesilate, Cefoxitin, Cyclosporine, Heparin, Ibuprofen, Intralipid, Levodopa, Methyldopa, Metronidazole, Phenylbutazone, Rifampicin, Tetracycline, Theophylline. PMID- 8704060 TI - There's no such thing as the amygdala. PMID- 8704061 TI - Blunted adrenocorticotropin and increased adrenal steroid response to human corticotropin-releasing hormone in Alzheimer's disease. AB - Previous studies suggest disturbances in the central regulation of the hypothalamic-pituitary-adrenocortical axis (HPA) in advanced Alzheimer's disease (AD). In this study the reactivity of the HPA axis was evaluated by stimulation with corticotropin-releasing hormone (CRH) of patients with mild to moderate AD. Twenty-three patients with AD (aged 61-88 yr) and 19 healthy elderly (aged 62-84 yr) received an intravenous bolus injection of human CRH (1 microgram/kg) at 3:00 PM. CRH-stimulated plasma ACTH levels were significantly lower in AD patients, while serum cortisol, dehydroepiandrosterone, and androstenedione responses relative to the amount of ACTH released were higher in AD patients. Significant correlations were found between low basal plasma ACTH levels and temporal lobe atrophy (p=0.02) and between peak plasma ACTH levels and hippocampal atrophy (p = 0.01). These findings suggest abnormalities at several levels of HPA axis in AD. PMID- 8704062 TI - Topography of the maximum of the amplitude of EEG frequency bands in dementia of the Alzheimer type. AB - The localization of the maxima of activity of 36 patients with dementia of the Alzheimer type and 36 age-matched controls was calculated after Fourier transformation of EEG data in six frequency bands (4 Hz steps between 1 and 24 Hz). Patients were divided into three groups of severity (BCRS mean < 3,9, 4.0 4.9, and > 4.9). Significant differences were found in the beta frequency band. In the distribution of the maxima of the activity, the maxima of patients were located more frontally than the maxima of controls. The alterations were staged dependent. The beta activity in EEG is said to be correlated with cognitive processes; the correlation between cognitive decline and decreasing beta activity in dementia of the Alzheimer type might support this assumption. Differences in the location of the maxima between patients and controls might allow for differentiation between both groups. PMID- 8704063 TI - EEG power modifications in obsessive-compulsive disorder during olfactory stimulation. AB - Temporal lobe electroencephalogram (EEG) activity was quantitatively analyzed in obsessive-compulsive disorder (OCD) when subjects are at rest and during a temporal lobe activating procedure, i.e., olfactory stimulation. At rest with eyes closed, delta-1 and alpha-2 power differences were evident in OCD patients as compared with normal controls. During olfactory stimulation, differences between patients and normal groups were detectable in the slower beta frequencies: Normal subjects showed a power increase, whereas OCD patients showed no modification or slight decrease. Our results support previous findings of temporal lobe EEG abnormalities in OCD patients with an abnormal pattern of response to a temporal lobe activating procedure. PMID- 8704064 TI - Brain potential changes after intranasal vs. intravenous administration of vasopressin: evidence for a direct nose-brain pathway for peptide effects in humans. AB - There is evidence that intranasal application of peptides is a way to circumvent the blood-brain barrier. This led us to compare the effects of arginine vasopressin (AVP) on event-related potentials (ERPs) in healthy men (n = 15) after intranasal and after intravenous (i.v.) administration. In a double-blind, crossover study, subjects received on three different occasions 20 IU of AVP intranasally (IN), 1.5 IU of AVP i.v., and saline solution. ERPs were recorded during the subject's performance on a auditory attention task. Plasma concentrations of vasopressin during task performance were enhanced after AVP, with the increase after i.v. administration of AVP exceeding that after AVP (p < 0.05). Intranasal administration of AVP substantially increased the P3 component of the ERP (p < 0.05). Intranasal administration of AVP substantially increased the P3 component of the ERP (< 0.01). By contrast, i.v. administration of AVP had no consistent effects on the ERP responses. In supplementary experiments as well, i.v. administration of lower doses of AVP (0.1 and 0.025 IU) did not affect the ERP. Plasma vasopressin concentrations after the 0.025 IU dose in these experiments were comparable to those after intranasal administration of 20 IU AVP. The results provide functional evidence that in the human brain effects of peptides like AVP may be facilitated after IN as compared to i.v. administration. PMID- 8704065 TI - Correlation between vasopressin baseline and TSH-blunting in depressives. AB - A blunted thyrotropin (TSH) response is a predictor of a good response to antidepressant drug treatment in depressives and neuroleptic treatment in paraphrenic patients (Larger et al 1986). The aim of the following study was to elucidate possible relationships between different endocrine systems and to shed light on the pathogenetic hypotheses of TSH-blunting. In order to evaluate especially hypothalamic activity in severe depression we were interested in the vasopressin system as another hormonal system underlying hypothalamic control. Thirty-four patients who met the criteria for major depression according to DSM III-R were subjected to the thyrotropin-releasing hormone (TRH) test. We also took baseline readings of the cortisol, neurophysinI (hNpI, reflecting vasopressin plasma levels), and neurophysinII (hNpII, reflecting oxytocin plasma levels) levels. Likelihood ratio tests were done with logistic regression models to analyze the phenomenon of TSH-blunting. We observed that the likelihood of a blunted TSH response increases with higher levels of hNpI and low levels of cortisol, but is unrelated to hNpII levels. PMID- 8704066 TI - Subjective memory complaints prior to and following electroconvulsive therapy. AB - Using the Squire Subjective Memory Questionnaire (SSMQ), depressed patients rated their memory functioning prior to a course of brief pulse, electroconvulsive therapy (ECT) within the 1 week following the course and 2 months later. Normal controls made similar ratings at comparable intervals. Prior to ECT, patients reported poorer memory functioning than controls. There was marked improvements in the patients' self-reports shortly following ECT, and at 2-month follow-up SSMQ scores were generally comparable in patients and controls. At all time points, the severity of depressive symptoms was strongly associated with patients' reports of memory dysfunction. SSMQ subscales ("depression" and "ECT" items) were not differentially sensitive to effects of ECT or depression. Relations between ECT treatment parameters and changes in patients' self evaluations only emerged after controlling for clinical state change. Shortly following ECT, there were no relations between SSMQ scores and objective measures of cognitive functioning. However, 2 months following ECT, there was a suggestion that greater retrograde amnesia for autobiographical memories was associated with self-rating of greater memory impairment. PMID- 8704068 TI - HPA axis function and temperament in depression: a negative report. AB - Positive correlations between scores on the Tridimensional Personality Questionnaire (TPQ) and plasma cortisol levels have led to the suggestion that temperament may be a major determinant of hypercortisolemia in depression. We had administered the Eysenck Personality Questionnaire (EPQ) to a group of 39 depressed patients who also underwent measurement of hypothalamic-pituitary adrenal (HPA) axis function. There were no significant correlations among the EPQ variables extraversion, neuroticism and psychoticism, and plasma cortisol levels, either pre or postdexamethasone, or with urinary free cortisol. Among a small group of six patients who completed the TPQ, there were similarly no significant correlations. PMID- 8704067 TI - Polymorphonuclear (PMN) elastase levels in depressive disorders. AB - The present study was carried out to determine polymorphonuclear (PMN) elastase levels and to explain whether or not altered levels may be a factor in depression. The patient group included a total of 68 patients with depression (32 with major depression, 36 with dysthymia). The control group included 40 volunteer, healthy subjects. WBC, erythrocyte sedimentation rate (ESR), immunogloblins (Ig A, IG G, IgM), and complements (C3 and C4) levels of the patient and control groups were determined. The mean PMN elastase levels, determined by immunoactivation method, in patients with major depression without and with melancholia, dysthymia and healthy subjects were found to be 99.5 (10.8) microgram(s)/L [arithmetic mean (SEM)], 289.0 (71.3) microgram(s)/L, 55.7(5.5) microgram(s)/L, and (47.3(2.6) microgram(s)/L, respectively. The mean PMN elastase level was found to be statistically higher in patients with depression, especially in patients with major depression, than that of healthy subjects. It was concluded that severe depression is associated with immunological and inflammatory alterations may be showed easily by PMN elastase measurements. PMID- 8704069 TI - Clozapine-induced myoclonus and generalized seizures. PMID- 8704070 TI - Impulsivity and neuroendocrine response to buspirone in bulimia nervosa. PMID- 8704071 TI - Olfactory identification deficits in relatives of Alzheimer's disease patients. PMID- 8704072 TI - Depression in women treated with a gonadotropin-releasing hormone agonist. PMID- 8704073 TI - Cerebrospinal fluid monoamine metabolism in a case of neuroleptic malignant syndrome improved by electroconvulsive therapy. PMID- 8704074 TI - Possible serotonin syndrome with trazodone addition to fluoxetine. PMID- 8704075 TI - Effect of nortriptyline on yohimbine-induced changes in heart rate variability. PMID- 8704076 TI - [Surfactant administration in AIDS]. PMID- 8704077 TI - [Prevention and treatment of postoperative nausea and vomiting with 5-HT3 receptor blockers]. AB - Since there continues to be a high incidence of postoperative nausea and vomiting associated with many types of surgery, and the standard antiemetics often do not achieve satisfactory results, there have been attempts to use the 5-HT3 antagonists. This group of substances is relatively new, but has already been used successfully as an antiemetic during chemotherapy. To date, results are on hand for four different 5-HT3 receptor blockers: ondansetron, tropisetron, granisetron and dolasetron. Applied intravenously, all four have been effective both in prophylaxis and also as therapy for postoperative emesis. Except for ondansetron, there is so far no definitely clear knowledge about the lowest possible effective dosage. The entire group is well tolerated: only occasional and minor side effects have been reported. Even though not all the hopes originally set in the 5-HT3 group of antagonists have been fulfilled, progress has nevertheless been achieved. Especially noteworthy points are a positive cost effectiveness relationship of these drugs and their appropriate use in case of the proper indications. PMID- 8704078 TI - [Is sepsis and septic shock due to a global oxygen insufficiency? A review in 2 parts. 2: Sepsis and lactate]. PMID- 8704079 TI - [Differential indications of non-opioid drugs for postoperative analgesia II. Quantification of the analgesic effect of a combination of metamizol plus diclofenac via patient-controlled analgesia]. AB - OBJECTIVE: In a previous study we investigated the analgesic efficacy of metamizol. After laparoscopic operations, in particular, the reduction of postoperative opioid requirements within the first 24 h after surgery attained clinical relevance (-67%). In the present study we investigated the analgesic efficacy of supplementary diclofenac. METHODS: 86 patients, scheduled for minor orthopaedic surgery, laparoscopic cholecystectomy or resection of the thyroid gland, participated in a doubleblind, randomised, placebo-controlled study. The setting was comparable to our previous study, apart from the supplementary administration of diclofenac. Before induction of anaesthesia, verum-treated patients received a diclofenac suppository (100 mg), in addition to metamizol (1 g/100 ml NaCl 0.9% intravenous over 15 min). These infusions were repeated at 6h and 12h. In addition to the third infusion, the patients received a further diclofenac suppository (100 mg). Cumulated doses of buprenorphine (PCA, patient controlled analgesia), pain scores (0-10), blood pressure, pulse and side effects were recorded during the first 6 h and again at 24 h. RESULTS: All verum-treated patients had significantly less pain immediately after surgery and required lower cumulated doses of buprenorphine during the first 24 h after operation (laparoscopic cholecystectomy -33%, minor orthopaedic surgery -73%, resection of thyroid gland -60%). CONCLUSIONS: Combination of metamizol and diclofenac cause a clinically relevant reduction in opioid requirements, in particular after minor orthopaedic surgery and resection of the thyroid gland. There is no need for supplementary diclofenac following laparoscopic surgery. PMID- 8704080 TI - [The Ulm Emergency Training Course. 2: Realization and acceptance of the emergency medicine practicum]. AB - The first part of this publication described the concept of the student course in emergency medicine at the University Hospital Ulm as part of a "vertical curriculum" of education in emergency medicine. METHODS. A training circuit (Fig. 1) was conceptualised using training manikins (i.e. BLS, intubation, megacode training, ATLS training), computer programmes (ECG-simulation) and other means of instruction to simulate twelve realistic situations. These practical skill sessions were linked with case presentation and lectures. Its topics were chosen depending on the prevalence, type and urgency. Separate guidelines for teachers and students were edited prior to the beginning of the course in 1994. RESULTS. Evaluation showed good acceptance by the students for most parts of the course except the ATLS skill station and CPR computer simulations (Table 5). The score of the national board examination (multiple-choice test) of the students from Ulm was nevertheless not better than the mean of all german examinees (72.1 to 71.7%). CONCLUSION. The concept of the course in emergency medicine is well accepted by our students. Results of the national examination were not improved, since the course aims at performance, skills and competence in emergency medicine and not at teaching factual knowledge that is measured by the MCQ examinations. PMID- 8704081 TI - [Optimizing a conventional documentation system on the surgical intensive care unit--a contribution to cost reduction and preventive quality management?]. AB - In Intensive Care Units (I.C.U.), the lack of staff and funds require the mobilisation of organisational reserves in order to ensure high-quality patient care. Traditional occupational-group organised documentation systems are burdened with lack of clarity, limited utilisation by the staff, insufficient information content and difficulties in synoptic patient monitoring. They cannot meet the demands of modern intensive-care medicine. At the inauguration of our Surgical I.C.U. in June 1992, an occupational-group oriented documentation system was introduced and put into operation. Because of negative repercussions on patient care, it was replaced by a patient-centered, conventional prescription and documentation system in April 1993. In April 1994, an evaluation of the patient centered system was carried out. We report on our initial problem analysis, the subsequent developmental and introductory phases, and the results after having used the system in our I.C.U. for one year. Data condensation, standardised data recording, as well as structured prescription, examination, assessment and decision processes, saved 730 working hours for medical and nursing staff per year, reduced the cost for documentation materials by 58% and improved the extent of data recording. In our experience, improving a conventional documentation system is a suitable instrument to support cost reduction and preventive internal quality management in the I.C.U. PMID- 8704082 TI - [Christian Bohr--a forgotten pioneer in respiratory physiology]. PMID- 8704083 TI - [Preoperative fasting and aspiration]. PMID- 8704084 TI - [Origins of preoperative fasting]. PMID- 8704085 TI - [Preoperative fasting and prevention of aspiration from the legal medicine viewpoint]. PMID- 8704086 TI - [Is traditional preoperative fasting still applicable?]. PMID- 8704087 TI - [Indications for emergency surgery]. PMID- 8704088 TI - [Conventional and new measures for prevention of aspiration]. PMID- 8704089 TI - [Medicamentous prevention of acid aspiration syndrome]. PMID- 8704090 TI - [Treatment of aspiration of gastric contents]. PMID- 8704091 TI - ["Treatment of total atelectasis of the left lung in severe ARDS with side separated ventilation and surfactant administration"]. AB - Severe thoracic trauma is always an important risk factor for the development of acute pulmonary failure. The course is often complicated by barotrauma or volutrauma. We report on a 48-year-old patient who was transferred to us nine days after a bicycle accident because of a severe disturbance of gas exchange and atelectasis of the left lung refractory to therapy. The left lung could not be ventilated even after separate artificial ventilation on each side with positive end expiratory pressure. After administration of surfactant (50 mg Exosurf per kg body weight) and continued separate artificial ventilation on each side, there was a complete re-expansion of the left lung with an increase of the arterial pO2 value from 65 mm Hg to 416 mm Hg with a FIO2 of 1,0 and a decrease of the intrapulmonary venous admixture from 34% to 12% within a few hours. The extravascular pulmonary fluid was unaffected by the administration of surfactant (200 ml solution). The administration of surfactant preparations may be a new therapeutic approach in treatment of ARDS patients. PMID- 8704092 TI - The cytokine network in sarcoidosis. PMID- 8704093 TI - Are interleukin-2 and interleukin-15 tumor promoting factors for human non hematopoietic cells? AB - Human normal non hematopoietic cells of mesenchymal and neuroectodermal origin may express functional IL-Rs. For instance, in these cell types, IL-2 can stimulate proliferation (endothelial, intestinal and nervous cells) or modify the expression of adhesion molecules (fibroblasts) or inhibit proliferation (bone marrow stromal cells). Therefore, some cytotoxic effects described during IL-2 biotherapy could be due to a direct interaction between IL-2 and non hematopoietic tissues. The expression of functional IL-2-R has also been reported in several human cell lines derived from solid tumors. In some instances IL-2 inhibits cell growth (head and neck, gastric and renal carcinomas), but in other tumors, growth stimulation and increased expression of markers of tumor progression have been reported (intestinal, breast, and lung carcinomas, gliomas, fibrosarcomas and melanomas). Additionally, secretion of biologically active IL-2 has been reported in some melanoma and breast cancer cell lines. Transcripts for the novel cytokine IL-15, which utilizes the beta and gamma chains of the IL2-R, have been found in melanoma cells and anti-IL-15 mAbs inhibit HLA class I expression in these cells. Therefore these cytokines may modify, inside a tumor, the behavior of both stromal and neoplastic cells. All these data may have important implications in our understanding of tumor host interactions and in future strategies of immunotherapy. PMID- 8704094 TI - Transmembrane and intracellular signalling by interleukin-4: receptor dimerization and beyond. AB - IL-4 is a paradigmatic example for hematopoietic cytokines of the four-helix bundle family, but offers some unusual features as well. Two receptor binding sites with different properties were identified in the molecule, which has allowed the development of antagonistic mutant proteins, and contributed to the now accepted concept that cytokines signal across membranes by crosslinking two receptor proteins. Intracellular signal transduction by IL-4 involves activation of Jak kinases and a pathway with homology to the insulin system, but not the ras/Raf-cascade. Receptor/ligand interactions are complex, because IL-4 and IL-13 share the main receptor component, and IL-4 can signal via a second type of receptor on some cells. PMID- 8704095 TI - Interleukin-1 and tumor necrosis factor-alpha in the Reed-Sternberg cells of Hodgkin's disease. Correlation with clinical and morphological "inflammatory" features. AB - We examined paraffin sections for the expression of interleukin-1 alpha, interleukin-1 beta and tumor necrosis factor-alpha, in 40 cases of Hodgkin's disease. Our purpose was to study the role of these cytokines in the "inflammatory" histological features and "B" symptoms in this disease. Immunohistochemistry with the avidin-biotin-peroxidase complex method was used. The findings were compared with those of 20 cases of non-Hodgkin's lymphomas and of 20 non-neoplastic lymphadenopathies. Evidence for EBV infection and myc and ras oncoproteins expression was also studied in these patients, but no correlation between any of these features and cytokine expression was found. We found a significant correlation between the expression of interleukin-1 beta and several "inflammatory" histological features, as well as between the expression of tumor necrosis factor-alpha and B symptoms and tumor bulk. The differential correlations between these major pro-inflammatory cytokines expression and the "inflammatory" manifestations in Hodgkin's disease are remarkable, considering the complexity of the cytokines composing the cytokine network involved in this disease. PMID- 8704096 TI - Regulation of cytokine release by alveolar macrophages treated with interleukin 4, interleukin-10, or transforming growth factor beta. AB - We evaluate the influence of IL-4, IL-10 and TGF-beta upon the release of IL-1 alpha, tumor necrosis factor-alpha (TNF-alpha), and IL-6 by lipopolisaccharide (LPS, 1 microgram/ml) stimulated alveolar macrophages (AM). IL-4 reduced TNF alpha release, in a dose dependent manner, to 62% and IL-1 alpha release to 42% of LPS-stimulated AM without IL-4. IL-6 release was also suppressed (61%), however, with a biphasic dose response curve. IL-10 suppressed LPS induced release of IL-alpha and TNF-alpha to approximately 50% of control without affecting IL-6 release. When used at high concentrations, TGF-beta achieved moderate reductions of TNF-alpha and IL-1 alpha release. Low concentrations of LPS (0.1 microgram/ml), allowed a dose-dependent TGF-beta-induced suppression of TNF-alpha-release to approximately 80% of control. The combinations of IL-4 and IL-10 was more effective in suppressing IL-6 release, although the suppression was only weak. Other combinations of cytokines revealed no synergistic inhibitory activities. Our data demonstrate that IL-1 alpha and TNF-alpha release by AMs is down regulated by IL-4, IL-10, and TGF-beta. IL-6 release, however, can only be suppressed to some extent by a combination of IL-4 and IL-10. PMID- 8704097 TI - Interleukin-10 pharmacokinetics in intact and nephrectomized mice. AB - The influence of kidney function on interleukin-10 (IL-10) pharmacokinetics was assessed by comparing the disappearance of IL-10 from the circulation in the intact and acutely nephrectomized mice over 1 h following a single bolus injection of E. coli-derived recombinant h IL-10 at 250 micrograms/kg i.v. The intact mice demonstrated a C(max) of 1,172 ng/ml and an AUC(tf) of 385 n g.h/ml. By comparison, there was a 4-fold elevation in C(max) and a 7-fold increase in AUC(tf) in the anephric mice. The serum IL-10 concentration at 1 h post-injection was 87 ng/ml in the intact vs 1,684 ng/ml in the anephric mice. The results imply that IL-10, in common with other cytokines, is eliminated through the kidney. PMID- 8704098 TI - In vivo effects of interleukin-1 receptor antagonist on hematopoietic bone marrow progenitor cells in normal mice. AB - The multiple effects of interleukin-1 (IL-1) on hematopoietic cells are mainly documented in disturbed hematopoiesis, but its production and participation during constitutive hematopoiesis are still unproven. To assess the involvement of IL-1 in the regulation of steady-state hematopoiesis in vivo, we have investigated the consequences of IL-1 receptor blockade by recombinant human IL-1 receptor antagonist (rhIL-1Ra) in normal CBA/H mice treated with two i.p. injections of rhIL-1Ra (2 x 50 micrograms/mouse) seventeen and two hours before sacrifice. The cellularity, the number of granulocyte-macrophage (CFU-GM), the number of erythroid (BFU-E) progenitor cells and the percentage of these cells in S phase of the cell cycle, as well as the morphologically recognizable cells in bone marrow were estimated. In peripheral blood, hematocrit, the number and differential count of nucleated cells, the number erythrocytes and the percentage of reticulocytes were determined. IL-1Ra treatment significantly reduced the number of femoral CFU-GM and BFU-E, while all the other analyzed parameters were not different from the level obtained in control, non-treated animals. These findings show that a number of bone marrow IL-1-responsive cells were affected by the IL-1 receptor blockade, indicating that the expression of IL-1 receptors and endogenous IL-1 secretion occur as part of constitutive hematopoiesis. PMID- 8704099 TI - Platelet-activating factor and haematopoiesis. XI. Platelet-activating factor has no effect on the production of interleukin-6 and tumor necrosis factor-alpha by human bone marrow stromal cells. AB - PAF is a phospholipid mediator of inflammation with stimulates IL-6 production by murine skin fibroblasts. Although PAF is present in human bone marrow, its role in haematopoiesis is unknown. We have assessed whether PAF stimulates IL-6 and TNF-alpha production by human bone marrow stromal cells (mostly fibroblast-like cells). We report that PAF (1 nM to 10 microM) has no effect on the synthesis of IL-6 and TNF-alpha by human bone marrow stromal cells. This difference may be due to the widely accepted concept "tissue-specific fibroblasts". The role of PAF in the regulation of human haematopoiesis remains to be elucidated. PMID- 8704100 TI - Cellular and molecular immunology of an important regulatory cytokine. New York 9 12, November 1995. PMID- 8704101 TI - Jenner and cytokines. PMID- 8704102 TI - Therapeutic plasma exchange in renal diseases. AB - Plasma exchange has been used extensively for over 2 1/2 decades to treat a variety of renal diseases. In this article, the scientific rationale for therapeutic plasma exchange in primary and secondary glomerulonephritis, thrombotic thrombocytopenic purpura and hemolytic uremic syndrome, myeloma cast nephropathy, and allograft rejection are reviewed. The clinical studies that evaluate its efficacy are summarized, with special emphasis on the results of randomized controlled trials, when available. Consensus plasma exchange regimens are presented for diseases in which there is evidence to support its use. PMID- 8704103 TI - Emerging concepts regarding B cells and autoantibodies in murine lupus nephritis. B cells have multiple roles; all autoantibodies are not equal. AB - Despite observations linking the severity of lupus nephritis to the quantity and location of glomerular immune deposits, it had been difficult to decipher the primary role of B cells and autoantibodies in this process. Newer technologies have provided the means to evaluate the roles of whole B cell populations and individual immunoglobulins in lupus lesions. In this review, recent advances in this area are summarized, with particular emphasis on work from the authors' laboratories. The results implicate a primary role for B cells and immunoglobulins in lupus nephritis, including glomerular, interstitial, and vascular lesions. Multiple antibody-ligand interactions participate in glomerular immune deposit formation in individuals with lupus nephritis. Recent evidence suggests that in situ formation of immune deposits by either cross-reactivity of autoantibodies with intrinsic glomerular antigens (i.e., anti-DNA antibodies with laminin) or direct interaction of autoantibodies with circulating autoantigens lodged within glomeruli (i.e., anti-DNA antibodies with histone/DNA). The predominant autoantibody-glomerular antigen interaction(s) in a given individual influences the principal location of immune deposition, which in turn influences the pathologic and clinical expression of disease. It is believed that these phenomena contribute to the phenotypic diversity commonly observed among individuals with lupus nephritis. Furthermore, these consequences are dependent on properties unique to both subsets of lupus autoantibodies and to their target antigen ligands within the glomerulus. Thus, the autoantibody variable or antigen binding region, along with the nature and location of the target glomerular antigen (or site of circulating antigen deposition), are influential in initiating these perturbations. PMID- 8704104 TI - Porphyria cutanea tarda in a patient on chronic ambulatory peritoneal dialysis. AB - Porphyria cutanea tarda is a disorder of heme biosynthesis resulting from a defect or deficiency in the enzyme uroporphyrinogen decarboxylase. Heme precursors accumulate in the blood, urine, stool, and skin, where exposure to sunlight results in the clinical manifestations. Porphyria cutanea tarda has been described in adult hemodialysis patients. The pathogenesis of porphyria cutanea tarda in this population is thought to be related to the inability of hemodialysis to adequately clear porphyrin precursors, resulting in increased precursor serum levels, precursor skin deposition, and subsequent clinical manifestations. A proper diagnosis of porphyria cutanea tarda in hemodialysis patients requires fractionation of serum porphyrins. Normalization of the porphyrin profile and reversal of the dermal manifestations require the withdrawal of hepatotoxic agents and the reversal of hepatic iron overload. A case of porphyria cutanea tarda in an adult ESRD patient treated with continuous ambulatory peritoneal dialysis is described. In this patient, the disease was related to elevated serum levels of phenytoin, which had been administered for seizure disorder. PMID- 8704105 TI - Urinary excretion of aquaporin-2 in humans: a potential marker of collecting duct responsiveness to vasopressin. AB - The vasopressin-sensitive water channel (aquaporin 2; AQP-2) mediates water transport across the apical plasma membrane of the renal collecting ducts and is excreted in human urine. This study presents the hypothesis that measurements of the AQP-2 excretion rate might be used as a marker of collecting-duct responsiveness to vasopressin, and therefore could be useful in the clinical evaluation of various water-balance disorders. This study presents information about the development of an antibody to human AQP-2, and measures the urinary excretion of AQP-2 by quantitative Western analysis. A standard curve of band densities was generated by using known quantities of the modified immunizing peptide to derive the amount of AQP-2 contained in aliquots of urine. AQP-2 urinary excretion changed with short-term alterations in hydration status produced either by water loading (76% decrease, P < 0.01) or by 3% sodium chloride (760% increase, P < 0.01). Steady-state 24-h urinary excretion of AQP-2 was 43 +/- 10 nmol/24 h (or 28.5 +/- 6.9 pmol/mg creatinine), and 20 +/- 6 nmol/24 h (or 18.3 +/- 7.9 pmol/mg creatinine) in men and women, respectively. Therefore, urinary AQP-2 excretion can be quantified by using Western analysis, and may serve as a marker of collecting-duct responsiveness to vasopressin in different physiologic settings. PMID- 8704106 TI - A low-affinity vasopressin V2-receptor gene in a kindred with X-linked nephrogenic diabetes insipidus. AB - In this study, a mutation in vasopressin Type 2 receptor (V2R) in a patient with hereditary nephrogenic diabetes insipidus (NDI) has been identified and characterized. The sequencing of the V2R gene from the patient revealed that there was a missense mutation (TAT to TGT) resulting in the substitution of 205Tyr for Cys in the putative third extracellular domain. The expression analysis in COS cells showed that the binding affinity of the mutant receptor (KD = 19.8 nM) for arginine vasopressin was much lower than that of the wild-type receptor (KD = 1.8 nM) so that intracellular cAMP production stimulated by arginine vasopressin was impaired in cells with the mutant V2R. From these results, it was concluded that the single amino-acid substitution of V2R is responsible for this familial disease. PMID- 8704108 TI - Contrast-enhanced dynamic magnetic resonance imaging of the rat kidney. AB - This study was designed to assess whether contrast-enhanced dynamic 1H magnetic resonance imaging (DMRI) can be used to detect the effects of the loop diuretic furosemide and the vasoactive peptide angiotensin II on tubular water reabsorption in the rat kidney. A bolus of gadolinium-DTPA-dimeglumine (Gd-DTPA) (0.025 or 0.1 mmol/kg) was used as a contrast agent. The signal intensity in the magnetic resonance images relative to the precontrast signal intensity (RSI) was assessed as a function of time in the cortex and medulla. In the cortex, no differences were observed between high and low bolus injection, and between different treatment groups and controls. In the medulla, RSI patterns were different between high and low bolus, with the high bolus showing lower RSI values, because of T2 shortening at high Gd-DTPA concentrations. No difference was observed between controls and angiotensin II-infused animals. This is in line with the finding that angiotensin II did not alter medullary water reabsorption, as evidenced by unchanged urine flow and osmolality compared with controls. Medullary RSI patterns during furosemide infusion differed markedly from controls, in a manner suggesting that a lower concentration of Gd-DTPA was present compared with controls. This agrees with the well-known inhibiting effect of furosemide on medullary water reabsorption. It was concluded that, with the method used, small concentration differences of Gd-DTPA in the cortex resulting from small changes of plus or minus 20% in tubular water reabsorption, previously found to be present by direct micropuncture measurements, cannot be detected in rats. However, large changes in renal concentrating ability do result in different RSI patterns in the medulla. PMID- 8704107 TI - Tyrosine phosphorylation of focal adhesion kinase (p125FAK): regulation by cAMP and thrombin in mesangial cells. AB - Stress fibers, composed of actin filaments, converge upon and associate with a number of proteins, including focal adhesion kinase (p125FAK), and integrin receptors to form areas of close contact between cells and the extracellular matrix referred to as focal adhesions. Treatment of mesangial cells with cAMP elevating agents causes a loss of focal adhesions, fragmentation of stress fibers, and decreased tyrosine phosphorylation of p125FAK. Thrombin reverses these effects of cAMP, and this model can be used to address some of the cellular mechanisms involved in regulating the loss and formation of focal adhesions. This study reports the effects of cAMP and thrombin on mesangial cell shape, distribution of actin, formation of stress fibers, and tyrosine phosphorylation of p125FAK. cAMP-treated cells display a condensed cell body with slender processes that traverse the area formerly covered by the cell. Addition of thrombin to these cells restores actin filaments (stress fibers) and increases tyrosine phosphorylation of p125FAK, and the cells resume a flattened morphology, even in the continued presence of cAMP-elevating agents. Peptides that mimic the tethered ligand portion of the thrombin receptor have the same effects on cell morphology and stress fiber formation as thrombin. In selected experiments, agents that disrupt either stress fibers (cytochalasin D) or microtubules (nocodazole; Sigma Chemical, St. Louis, MO) were used to examine the role of these cytoskeletal elements in thrombin-induced restoration of focal adhesions. Cytochalasin D blocked the ability of thrombin to restore focal adhesions and phosphorylate p125FAK. The effects of nocodazole, an agent that destabilizes microtubules (but which has no known receptor), are very similar to those of thrombin. The findings discussed in this study indicate that thrombin can modulate the formation of focal adhesions. The organization of stress fibers and microtubules is apparently intimately related to the phosphorylation of p125FAK and can be modulated by soluble receptor agonists such as thrombin or via altered polymerization of microtubules. PMID- 8704109 TI - Intestinal lipid absorption in the nephrotic rat. AB - There is limited data on intestinal lipid absorption in the nephrotic syndrome. This study investigated whether the efficiency of intestinal lipid absorption is altered in nephrotic lymph-fistula rats. The nephrotic syndrome was induced in nine Sprague-Dawley rats by an i.v. injection of puromycin aminonucleoside in saline; seven control rats received saline only. At 10 to 14 days after injection, the main intestinal lymph duct was cannulated for collection of lymph. The duodenum was also cannulated and a fasting saline-glucose solution was infused overnight at 3 mL/h. The next day, the infusate was changed to a lipid emulsion that contained (14C) cholesterol and (3H)triglyceride (triolein) that was infused at 3 mL/h for 8 h. During the last hour of fasting and during the lipid infusion, lymph flow in the Nephrotic group averaged 0.6 mL/h higher than the Control group (P = 0.02). No significant differences were found between the two groups in recovery of infused radioactive cholesterol (P = 0.37) or triglyceride (P = 0.38) from the gastrointestinal lumen, small intestinal mucosa, or lymph. Lymphatic output of chemically measured cholesterol was also similar (P = 0.96). These results suggest that mucosal uptake and lymphatic output of cholesterol and triglyceride are not altered in the nephrotic syndrome. PMID- 8704110 TI - Salt restriction inhibits renal growth and stabilizes injury in rats with established renal disease. AB - Salt restriction inhibits renal growth and stabilizes injury in rats with established renal disease. Male Munich-Wistar rats that underwent right nephrectomy and segmental infarction of two thirds of the left kidney were fed standard chow for 4 wk and then randomly assigned to ingest standard or low-salt chow for an additional 4 wk. Four wk after ablation, rats had systemic hypertension, proteinuria, and glomerular sclerosis. The prevalence of sclerosis, protein excretion rate, and glomerular volume increased between the fourth and eighth week in rats that were fed standard chow, however, in rats that were fed low-salt chow, the increase in glomerular volume and development of further glomerular sclerosis was prevented whereas the protein excretion rate actually declined. Micropuncture studies performed 8 wk after ablation revealed that the glomerular hydraulic pressure was elevated in remnant kidneys and was not affected by salt restriction. This study demonstrates that dietary salt restriction can prevent further glomerular injury and reduce proteinuria even when instituted in rats with established renal disease. These findings are also consistent with the hypothesis that glomerular hypertrophy promotes injury in this model of hypertension and progressive renal disease. PMID- 8704111 TI - Active kallikrein response to changes in sodium-chloride intake in essential hypertensive patients. AB - To evaluate the behavior of active kallikrein excretion in salt-sensitive and salt-resistant hypertensive patients during changes in sodium-chloride (NaCl) intake, 61 male, nonobese, nondiabetic outpatients affected by uncomplicated essential hypertension were given a diet that contained 140 mmol NaCl per day for 2 wk. Patients then received either a low- (20 mmol NaCl/day) or a high- (320 mmol NaCl/day) sodium diet for 2 wk, according to a randomized, double-blind, cross-over protocol. Hypertensive patients were classified as salt sensitive when their diastolic blood pressure rose by at least 10 mm Hg after the high-sodium diet, and decreased by at least 10 mm Hg after the low-sodium diet, considering as baseline blood pressure values those that were taken at the end of the 140 mmol NaCl/day intake period. The remaining patients were classified as salt resistant or, when diastolic blood pressure increased by 10 mm Hg or more after low-sodium intake, as counter-regulating. Twenty-three patients were therefore classified as salt sensitive, 28 as salt resistant, and 10 as counter-regulating. The baseline active kallikrein excretion was significantly lower (P < 0.0001) in salt-sensitive (0.62 +/- 0.31 U/24 h) patients than in salt-resistant (1.39 +/- 0.44 U/24 h) and counter-regulating patients (1.27 +/- 0.38 U/24 h). Surprisingly, the kallikrein response to changes in sodium intake was similar in all subgroups, although enzyme excretion was always at the lowest level in salt sensitive hypertensive patients. This latter group also showed the highest plasma atrial natriuretic peptide levels (28.2 +/- 8.5 fmol/mL, P < 0.0001 versus salt resistant and counter-regulating patients), and the greatest peptide increment with sodium load (P < 0.0001 versus salt-resistant and counter-regulating patients). Counter-regulating patients showed the steepest increase in plasma renin activity (from 0.24 +/- 0.18 to 0.83 +/- 0.21 ng/L per s, P < 0.001) and decrease of plasma atrial natriuretic peptide (from 26.1 +/- 6.3 to 6.8 +/- 3.1 fmol/mL, P < 0.001) when switched from a high to a low-sodium intake. In conclusion, salt-sensitive hypertensive patients excrete less active kallikrein than do salt-resistant and counter-regulating patients, but maintain a normal enzyme response to changes in dietary sodium intake. The exaggerated response of atrial natriuretic peptide to high-sodium intake that was observed in the same patients could be compensating for an impaired renal capability to excrete a sodium load. PMID- 8704112 TI - Relationship of Na-K-ATPase inhibitors to blood-pressure regulation in continuous ambulatory peritoneal dialysis and hemodialysis. AB - Inhibitors of sodium-potassium-activated adenosine triphosphatase (Na-K-ATPase) have been implicated in the pathogenesis of hypertension. In the study presented here, an attempt was made to determine whether differences in the plasma levels and the removal rates of high-molecular weight (HMW) and low-molecular weight (LMW) forms of Na-K-ATPase inhibitors might relate to blood-pressure control in hemodialysis (N = six ultrafiltered and N = six non-ultrafiltered) and CAPD (N = six long-term and N = five short-term) patients. The latter group was studied before the initiation of continuous ambulatory peritoneal dialysis (CAPD) and 2 wk after starting the treatment. The mean blood pressure was significantly reduced after dialysis in the nonultrafiltered hemodialysis group and in both CAPD groups. Plasma levels of both HMW and LMW inhibitors were found to be elevated before dialysis in all patients and were modified only slightly after dialysis. Irrespective of whether ultrafiltration was utilized in hemodialysis patients and despite significant losses of both HMW and LMW inhibitors into CAPD effluent. Because CAPD effluent was found to contain vasopressors that were not exclusively Na-K-ATPase inhibitors, losses of these other vasopressors may contribute to improved blood-pressure control in CAPD in contrast to hemodialysis. PMID- 8704113 TI - Multicenter clinical validation of an on-line monitor of dialysis adequacy. AB - Quantitation of hemodialysis by measuring changes in blood solute concentration requires careful timing when taking the postdialysis blood sample to avoid errors from postdialysis rebound and from recirculation of blood through the access device. It also requires complex mathematical interpretation to account for solute disequilibrium in the patient. To circumvent these problems, hemodialysis can be quantified and its adequacy assessed by direct measurement of the urea removed in the dialysate. Because total dialysate collection is impractical, an automated method was developed for measuring dialysate urea-nitrogen concentrations at frequent intervals during treatment. A multicenter clinical trial of the dialysate monitoring device, the Biostat 1000 (Baxter Healthcare Corporation, McGaw Park, IL) was conducted to validate the measurements of urea removed, the delivered dialysis dose (Kt/V), and net protein catabolism (PCR). The results were compared with a total dialysate collection in each patient. During 29 dialyses in 29 patients from three centers, the paired analysis of urea removed, as estimated by the dialysate monitor compared with the total dialysate collection, showed no significant difference (14.7 +/- 4.7 g versus 14.8 +/- 5.1 g). Similarly, measurements of Kt/V and PCR showed no significant difference (1.30 +/- 0.18 versus 1.28 +/- 0.19, respectively, for Kt/V and 42.3 +/- 15.7 g/day versus 52.2 +/- 17.4 g/day for PCR). When blood-side measurements during the same dialyses were analyzed with a single-compartment, variable-volume model of urea kinetics, Kt/V was consistently overestimated (1.49 +/- 0.29/dialysis, P < 0.001), most likely because of failure to consider urea disequilibrium. Because urea disequilibrium is difficult to quantitate during each treatment, dialysate measurements have obvious advantages. The dialysate monitor eliminated errors from dialysate bacterial contamination, simplified dialysate measurements, and proved to be a reliable method for quantifying and assuring dialysis adequacy. PMID- 8704114 TI - The effect of membrane biocompatibility on plasma beta 2-microglobulin levels in chronic hemodialysis patients. AB - Several studies have shown that patients who have been dialyzed with high-flux biocompatible membranes have a lower plasma level of beta 2-microglobulin and a lower incidence of amyloid disease compared with patients who have been dialyzed with low-flux bioincompatible membranes. However, because high-flux membranes are associated with significant dialytic removal of beta 2-microglobulin, the specific role of membrane biocompatibility in influencing the rate of increase of beta 2-microglobulin has not been previously determined. This study investigated the effect of biocompatibility on the rate of increase of plasma levels of beta 2 microglobulin in 159 new hemodialysis patients from 13 dialysis centers (ten centers affiliated with Dallas Nephrology Associates and three with Vanderbilt University Medical Center) by using two low-flux membranes with widely different biocompatibilities. These patients were prospectively randomized to be dialyzed with either a low-flux biocompatible membrane or a low-flux bioincompatible membrane. Plasma beta 2-microglobulin levels were measured at 0, 3, 6, 9, 12, and 18 months. Sixty-six patients completed the 18-month study. Plasma beta 2 microglobulin increased in all patients; however, the increase was not significantly different from baseline at any time point in the group that used the biocompatible membrane. In this group, beta 2-microglobulin increased from (mean +/- SD) 27.8 +/- 14.8 mg/L to 34.0 +/- 10.0 mg/L at 18 months (P = not significant), and the mean increase at 18 months was 2.6 +/- 14.7 mg/L. In contrast, the increase in plasma beta 2-microglobulin level in the bioincompatible membrane group became significant in Month 6 when the levels had increased from a baseline of 24.8 +/- 9.6 mg/L to 29.5 +/- 12.2 mg/L (P < 0.001); these increases continued to be significant until Month 18, when serum beta 2 microglobulin reached 36.8 +/- 13.9 mg/L with an average increase of 11.8 +/- 11.2 mg/L (P < 0.0001). The higher rate of plasma B2-microglobulin increase in the group that had been dialyzed with the bioincompatible membrane was also evident when only patients who had completed the study were analyzed. There were no significant differences in the actual level of beta 2-microglobulin or in residual renal function between the two groups during the 18 months of the study. It was concluded that over a period of 18 months, the use of biocompatible membranes, even in the low-flux configuration, is associated with a significantly slower increase in plasma beta 2-microglobulin, independent of the influence of residual renal function. PMID- 8704115 TI - Plasma lipopolysaccharide binding protein and bactericidal/permeability increasing factor in CRF and HD patients. AB - The recent characterization of lipopolysaccharide binding protein (LBP) and bactericidal/permeability increasing factor (BPI) have provided the opportunity to examine the natural factors that regulate cytokine production in response to endotoxin in patients on hemodialysis (HD). Whole blood was collected in EDTA from 28 undialyzed patients with chronic renal failure (undialyzed CRF), 36 patients on chronic HD (HD) and 15 healthy controls, and plasma levels of LBP and BPI were measured by a sandwich ELISA. Plasma LBP levels in undialyzed patients with CRF (P = 0.04) and patients on HD (P = 0.01) were significantly higher than those in healthy controls, but not significantly different from each other. Plasma BPI levels in undialyzed patients with CRF and patients on HD were not significantly different from those in healthy controls. There was no correlation between serum creatinine and plasma levels of either LBP or BPI. Peripheral blood mononuclear cells (PBMC) were harvested from healthy volunteers by FLcoll-Hypaque separation, and 0.125 mL of 10 x 10(6)/mL suspensions were incubated with 0.125 mL of test plasma (containing different LBP/BPI ratios) and 0.25 mL of RPMI, containing 1 ng/mL of endotoxin, for 24 h at 37 degrees C. Samples were subjected to three freeze-thaw cycles, and total interleukin-1 receptor antagonist (IL-1Ra) or interleukin-1 alpha (IL-1 alpha) production was measured by a specific non crossreactive RIA. The results of this study showed: (1) IL-1Ra production by endotoxin-stimulated PBMC incubated with pooled plasma from HD patients with LBP/BPI ratios of 11 x 10(2), 167 x 10(2), 379 x 10(2), and 778 x 10(2), respectively was 1466 +/- 195 pg, 3105 +/- 462 pg, 8179 +/- 1020 pg, and 4770 +/- 1185 pg (P < 0.001); (2) Paired plasma collected before dialysis (predialysis) and at 15 min after the start of dialysis (15 minute) with cellulose membranes showed a negligible change in plasma LBP levels (-3 +/- 5%), but a 6681 +/- 1788% increase in plasma BPI levels. Consequently, compared with predialysis plasma, there was a 35 +/- 6% decrease in endotoxin-stimulated IL-1 alpha production by PBMC incubated with plasma drawn at 15 min (P = 0.001); (3) Compared with the PBMC incubated with predialysis plasma from HD patients, there was a 39 +/- 5%, 53 +/- 5%, and 60 +/- 5% decrease in endotoxin-stimulated IL-1 alpha production in the presence of 1 ng/ mL, 10 ng/mL, or 1 microgram/mL of recombinant BPI, respectively (P < 0.003). These results suggest that the plasma LBP:BPI ratio could influence cytokine production in response to bacterial endotoxin; the high LBP:BPI ratios observed in patients with chronic renal failure probably imparts an increased susceptibility to endotoxin-stimulated cytokine production; and natural or pharmacological increases in plasma BPI levels and the consequent decrease in LBP:BPI ratios could attenuate this susceptibility to endotoxin stimulated cytokine production. PMID- 8704116 TI - Pharmacokinetics and efficacy of pulse oral versus intravenous calcitriol in hemodialysis patients. AB - Because intravenous (iv) calcitriol has greater bioavailability than oral calcitriol, it may be more efficacious in suppressing parathyroid hormone (PTH) secretion. In this study, the pharmacokinetics and efficacy of pulse oral and i.v. calcitriol were compared. Patients were randomized to receive 2 micrograms of i.v. or oral calcitriol after each dialysis. Two pharmacokinetic studies (PK1, PK2) were performed 10 days apart, during which the patients received calcitriol after each dialysis. Calcitriol bioavailability was determined from the area under the curve (AUCtime interval (hours) in pg/mL per h). After the PK phase, PTH was lowered to < 200 pg/mL by titrating calcitriol to a maximum of 12 micrograms/wk over 4 wk. Calcitriol was then maintained for another 18 wk unless serum calcium exceeded 11.5 mg/dL or Ca x P product exceeded 70; when these limits were reached, calcitriol was held and then restarted at a lower dose. After i.v. administration, peak serum calcitriol exceeded that achieved orally but by 1 h, calcitriol levels were similar. The AUC0-0.5 (105 +/- 12, i.v.; 9 +/- 4, oral) and AUC0.5-1 (68 +/- 6, i.v.; 30 +/- 7, oral) were higher with i.v. (P < 0.05), but cumulative AUC0-48 did not differ. Individual t1/2 values ranged from 10 to 129 h for PK1 and from 10 to 50 h for PK2. The t1/2 for oral calcitriol was 38 +/- 14 h for PK1 and 30 +/- 4 h for PK2 (not significant (NS)). The t1/2 for i.v. calcitriol was 26 +/- 5 h for PK1 and 19 +/- 3 h for PK2 (NS, PK1 versus PK2 and oral versus i.v.). When the PK1 oral and i.v. data were combined, the mean t1/2 was 32 +/- 7 h whereas the t1/2 for PK2 (oral and i.v.) was 22 +/- 3 h (P < 0.05). Baseline PTH levels were 510 +/- 90 pg/mL and 499 +/- 79 pg/mL, oral and i.v., respectively. Serum PTH level at 22 wk was not different between oral and i.v. groups, 153 +/- 38 pg/mL and 214 +/- 124 pg/mL in i.v. (NS). The percentage of PTH suppression was 66 +/- 7.4% in the oral group and 69 +/- 12% in the i.v. group (NS). A major degree of serum iPTH suppression occurred during the initial 4 wk of treatment, concomitant with a rise in serum calcium levels. Adverse effects were similar between groups, as were the average dosages of calcitriol and phosphate binders. In conclusion, the efficacy of intravenous and pulse oral calcitriol were similar in hemodialysis patients with secondary hyperparathyroidism. The early rise in serum calcium levels observed with treatment may have contributed significantly to the suppression of serum iPTH levels. The difference in bio-availability between the different routes does not have a clinically apparent effect. The t1/2 varied widely among individuals, whereas exposure to calcitriol may decrease the t1/2. PMID- 8704117 TI - Calcitriol and the parathyroid hormone-ionized calcium curve: a comparison of methodologic approaches. AB - Investigations of the effects of calcitriol on the inverse sigmoidal relationship between parathyroid hormone (PTH) and ionized calcium (iCa) have yielded contradictory conclusions, possibly because of variations in experimental and analytical approaches. To clarify the existing literature, PTH-iCa curves were constructed by inducing hypo- and hypercalcemia through alterations in dialysate calcium concentration in eight hemodialysis patients with mild to moderate secondary hyperparathyroidism. The effects of low (metabolic acidosis) versus normal bicarbonate dialysis were compared before and after 4 wk of intravenous calcitriol, in a cross-over design. The PTH-iCa curves were primarily evaluated by using a four-parameter model. In addition, a variety of alternative published analytic approaches were examined and PTH-iCa curve slopes were further evaluated after normalization by (maximum - minimum PTH). The latter partially corrects for gland mass and cell-secretory capacity, and therefore yields a purer measure of the sensitivity of secretory activity to changes in iCa. The results of the study indicate that calcitriol decreased basal, maximal, and minimal PTH and non normalized slope (all P < 0.05), but did not affect set point or normalized slope, independent of the specific analytic approach. Acute metabolic acidosis did not affect the PTH-iCa curve. Thus, intravenous calcitriol appears to decrease parathyroid gland functional mass, as reflected by decreases in maximal and minimal PTH levels, but does not affect the sensitivity of the parathyroid gland to changes in iCa, as set point and normalized slope were unaffected. PMID- 8704118 TI - Plasma total versus bone alkaline phosphatase as markers of bone turnover in hemodialysis patients. AB - Plasma total versus bone alkaline phosphatase as markers of bone turnover in hemodialysis patients. Plasma bone-specific alkaline phosphatase (bAP) has been demonstrated to be more reliable than total alkaline phosphatases (tAP) in providing information about bone turnover in patients with metabolic bone diseases. This study surveyed 42 hemodialysis patients who underwent a systematic transiliac bone biopsy for histomorphometry study. Plasma bAP was determined by using a new immunoassay (Tandem-R Ostase, Hybritech, Liege, Belgium). Plasma bAP values were compared with those of two other plasma markers of bone metabolism, namely tAP and intact parathyroid hormone (iPTH), for the correlations with bone histomorphometric parameters. Patients with high-turnover bone disease (HTBD) (N = 32) had significantly higher plasma bAP levels than patients with normal or low bone turnover (N/LTBD) (N = 10) (66.9 +/- 63.5 ng/mL versus 10.8 +/- 4.2 ng/mL, respectively). Bone formation and resorption were highly correlated in these patients, and plasma bAP levels were positively correlated with bone resorption parameters, including osteoclast surface (r = 0.39, P < 0.0001) and osteoclast number/mm2 (r = 0.36, P < 0.001), and with bone formation parameters, osteoblast surface (r = 0.50, P < 0.005), and bone formation rate (r = 0.91, P < 0.0001). The bone formation rate was better correlated with plasma bAP levels than with either plasma tAP or iPTH concentrations. Plasma bAP level equal or higher than 20 ng/mL, either alone or combined with plasma iPTH of 200 pg/mL, had the highest sensitivity, specificity, and predictability values for the diagnosis of high turnover bone disease, and formally excluded patients with normal or LTBD. In conclusion, plasma bAP can be measured with a reliable immunoassay in hemodialysis patients. It represents a highly sensitive and specific biochemical marker of skeletal remodeling in these patients. Therefore, both serum iPTH and bAP are complementary in diagnoses of the type of renal osteodystrophy. PMID- 8704119 TI - Dietary fish oil in renal transplant recipients treated with cyclosporin-A: no beneficial effects shown. AB - This study aimed to determine whether dietary supplementation with fish oil has a beneficial effect on graft function and the incidence of rejection in renal allograft recipients treated with cyclosporin A (CsA). Renal function, blood pressure, the incidence of acute rejection episodes, graft survival, and renal histology and immunochemistry were investigated. In a randomized, placebo controlled, double-blind trial, groups of 25 recipients of primary cadaveric renal allografts who had been treated with CsA took fish oil (30% C20:5 omega-3 and 20% C22:6 omega-3) or coconut oil (63% C8:0 and 36% C10:0) at 6 g/day for 3 months. There were no differences between the two patient groups with regard to HLA matching, panel-reactive antibody titers, or the demographic characteristics of donors or recipients. The GFR and effective RPF were determined at 1, 3, and 12 months after transplantation by simultaneous measurement of (125I-)iothalamate and (131I-)hippuran clearances. At 1 yr after transplantation, patients treated with fish oil showed better renal function than did the control patients, but this difference was not statistically significant. Blood pressure and antihypertensive drug use were similar in both groups. The number of rejection episodes was also similar, and renal histopathological and immunohistochemical studies showed no significant differences between the fish-oil group and the control patients. It is concluded that fish oil, at a dose of 6 g/day, has no beneficial effect after renal transplantation within the time scale of the study. PMID- 8704120 TI - Arabinogalactan proteins and plant differentiation. PMID- 8704121 TI - The starch phosphorylase gene is subjected to different modes of regulation in starch-containing tissues of potato. AB - Analysis of the levels of starch phosphorylase mRNA and its product in the various organs of the potato plant indicates that the gene is differentially regulated, leading to a high accumulation of the gene product in tubers. The amount of phosphorylase transcripts synthesized in nuclei isolated from tubers and leaves indicates that the difference in the steady-state levels of phosphorylase mRNA in these organs can be explained by different rates of initiation of transcription. However, while rates of initiation of transcription are similar in tubers and stems, the steady-state level of phosphorylase mRNA is much lower in the stem. Transgenic potato plants expressing the beta glucuronidase (GUS) gene under the control of 5'-flanking sequences of the phosphorylase gene exhibited high levels of GUS activity in petioles, stems, stolons, tubers and roots, but low levels in leaves. This confirms the results of transcription assays observed for leaves, stems and tubers, and indicates that accumulation of phosphorylase mRNA in stems and tubers is not controlled solely by transcription initiation. Finally, histochemical analysis for GUS activity in transgenic potato plants suggests that transcription of the phosphorylase gene predominantly occurs in starch-containing cells associated to vascular tissues, and suggests a role for starch phosphorylase in the mobilization of starch stored along the translocation pathway. PMID- 8704122 TI - High-resolution mapping of repetitive DNA by in situ hybridization: molecular and chromosomal features of prominent dispersed and discretely localized DNA families from the wild beet species Beta procumbens. AB - Members of three prominent DNA families of Beta procumbens have been isolated as Sau3A repeats. Two families consisting of repeats of about 158 bp and 312 bp are organized as satellite DNAs (Sau3A satellites I and II), whereas the third family with a repeat length of 202 bp is interspersed throughout the genome. Multi colour flourescence in situ hybridization was used for physical mapping of the DNA families, and has shown that these tandemly organized families occur in large heterochromatic and DAPI positive blocks. The Sau3A satellite I hybridized exclusively around or near the centromeres of 10, 11 or 12 chromosomes. The Sau3A satellite family I showed high intraspecific variability and high-resolution physical mapping was performed on pachytene chromosomes using differentially labelled repeats. The physical order of satellite subfamily arrays along a chromosome was visualized and provided evidence that large arrays of plant satellite repeats are not contiguous and consist of distinct subfamily domains. Re-hybridization of a heterologous rRNA probe to mitotic metaphase chromosomes revealed that the 18S-5.8S-25S rRNA genes are located at subterminal position on one chromosome pair missing repeat clusters of the Sau3A satellite family I. It is known that arrays of Sau3A satellite I repeats are tightly linked to a nematode (Heterodera schachtii) resistance gene and our results show that the gene might be located close to the centromere. Large arrays of the Sau3A satellite II were found in centromeric regions of 16 chromosomes and, in addition, a considerable interspersion of repeats over all chromosomes was observed. The family of interspersed 202 bp repeats is uniformly distributed over all chromosomes and largely excluded from the rRNA gene cluster but shows local amplification in some regions. Southern hybridization has shown that all three families are specific for genomes of the section Procumbentes of the genus Beta. PMID- 8704123 TI - Post-termination-induced and hormonally dependent expression of low-molecular weight heat shock protein genes in Douglas fir. AB - We have isolated and sequenced two cDNA clones (PM 18.2A; PM 18.2B) from Douglas fir (Pseudotsuga menziesii (Mirb.) Franco) which encode for the low-molecular weight heat shock proteins (LMW HSPs) of 18.2 kDa. The predicted amino acid sequences of the two Douglas fir proteins are 97.5% identical. A phylogenetic tree of class I LMW HSPs showed that the PM LMW HSPs are found within a subgroup consisting exclusively of dicot species indicating that class I LMW HSPs evolved from a common ancestor predating the divergence of gymnosperms and angiosperms. Northern blots of RNA from dry, imbibed, stratified and germinated seeds revealed a notable induction of LMW HSP transcripts during post-germination and early seedling growth. Unlike previous reports, the expression of these HSPs appears to be primarily restricted to seedlings as mRNA transcripts were detected at very low levels during seed development and desiccation. Maximum induction of LMW HSPs in seedlings occurred during heat shock treatment at 38-40 degrees C, whereas cold shock or wounding failed to induce HSP transcripts. The transcription of HSP genes is up regulated by GA, MeJA and auxin and is down regulated by ABA. Methyl jasmonate treatment induced expression of these genes in dormant seeds of Douglas fir. The expression of class I cytoplasmic LMW HSPs in seedlings and their regulation by plant growth regulators suggests specific roles in plant development other than desiccation tolerance. PMID- 8704124 TI - Calcium-dependent protein kinase gene expression in response to physical and chemical stimuli in mungbean (Vigna radiata). AB - Protein kinases are important in eukaryotic signal transduction pathways. In this study we designed degenerate oligonucleotides corresponding to two conserved regions of protein kinases and using the polymerase chain reaction (PCR) have amplified a 141 bp fragment of DNA from mungbeans (Vigna radiata Rwilcz cv. Berken). Sequence analysis of the PCR products indicates that they encode several putative protein kinases with respect to their identity with other known plant protein kinases. Using one of the six fragments (CPK3-8), we isolated a 2022 bp cDNA (VrCDPK-1) from a Vigna radiata lambda gt11 library. VrCDPK-1 has a 96 bp 5' untranslated region and a 465 bp 3'-untranslated region and an open reading frame of 1461 bp. VrCDPK-1 contains all of the conserved regions commonly found in calcium dependent protein kinases (CDPK). VrCDPK-1 shares 24 to 89% sequence identity with previously reported sequences for plant CDPKs at the protein level. Southern analysis revealed the presence of several copies of the CDPK gene. VrCDPK-1 expression was stimulated when mungbean cuttings were treated with CaCl2, while treatment with MgCl2 had no effect. We are reporting for the first time a CDPK gene in mungbean which is inducible by mechanical strain. Cuttings treated with indole-3-acetic acid (IAA) or subjected to salt stress showed an increase in VrCDPK-1 expression. There was a dramatic stimulation in VrCDPK-1 expression 6 h after cuttings were treated with cycloheximide. PMID- 8704125 TI - Molecular cloning, in vitro expression and characterization of a plant squalene synthetase cDNA. AB - Squalene synthetase (farnesyl-diphosphate:farnesyl-diphosphate farnesyltransferase, EC 2.5.1.21) catalyzes the first committed step for sterol biosynthesis and is thought to play an important role in the regulation of isoprenoid biosynthesis in eukaryotes. Using degenerate oligonucleotides based on a conserved region found in yeast and human squalene synthetase genes, a cDNA was cloned from the plant Nicotiana benthamiana. The cloned cDNA contained an open reading frame of 1234 bp encoding a polypeptide of 411 amino acids (M(r) 47002). Northern blot analysis of poly(A)+ mRNA from N. benthamiana and N. tabacum cv. MD609 revealed a single band of ca. 1.6 kb in both Nicotiana species. The identity and functionality of the cloned plant squalene synthetase cDNA was further confirmed by expression of the cDNA in Escherichia coli and in a squalene synthetase-deficient erg9 mutant of Saccharomyces cerevisiae. Antibodies raised against a truncated form of the protein recognized an endogenous plant protein of appropriate size as well as the full-length bacterially expressed protein as detected by western analysis. Comparison of the deduced primary amino acid sequences of plant, yeast, rat and human squalene synthetase revealed regions of conservation that may indicate similar functions within each polypeptide. PMID- 8704126 TI - The expression pattern of alfalfa flavanone 3-hydroxylase promoter-gus fusion in Nicotiana benthamiana correlates with the presence of flavonoids detected in situ. AB - Flavanone 3-hydroxylase is an enzyme acting in the central part of the flavonoid biosynthesis pathway. It is generally encoded by a single gene and seems to have a key position for the regulation in this pathway. These two features make a single f3h promoter-gus fusion a suitable tool to study both the f3h expression and the regulation of this pathway. We present here the spatial and temporal analysis of the expression of an alfalfa flavanone 3-hydroxylase (f3h) promoter gus fusion introduced into Nicotiana benthamiana. The Medicago sativa (alfalfa) f3h promoter directed gus expression in flowers, stems, leaves and roots. In flowers, GUS activity was observed in pollen grains, in ovules, in ovary placenta and in the epidermis, medullary parenchyma, trichomes and second cortical cellular layer surrounding the vascular bundles of the peduncle. In stems, GUS activity was detected at the same places as in the peduncle except for the medullary parenchyma. In roots, we found GUS staining in root hairs, epidermis and in the vascular bundles of the elongated zone. Finally, in leaves, the f3h promoter expressed essentially in the stalk cells of the multicellular trichomes. The expression pattern of the f3h-gus fusion was correlated to the presence of flavonoids in situ. These data indicate that this construct can be very useful to study factors controlling the production of flavonoids. PMID- 8704127 TI - Expression of the type 2 metallothionein-like gene MT2 from Arabidopsis thaliana in Zn(2+)-metallothionein-deficient Synechococcus PCC 7942: putative role for MT2 in Zn2+ metabolism. AB - Zn2+ proteins pervade metabolism and are essential for gene expression. However, no proteins have been ascribed the central roles of Zn2+ donation to, or removal from, metalloproteins, or Zn2+ storage in vegetative plant tissue. In animals, such functions have been proposed for metallothioneins. Plants contain multiple metallothionein-like genes but their predicted products, which differ significantly from animal metallothioneins, remain to be isolated from vegetative tissue and their roles are uncertain. The type 2 metallothionein-like gene from Arabidopsis, MT2, was expressed under the control of Zn2+-responsive elements derived from the cyanobacterial metallothionein divergon, smt. Zn2+-dependent expression of MT2 transcripts in Synechococcus PCC 7942 was confirmed by northern analysis. The Arabidopsis MT2 gene partly complemented Zn2+ hypersensitivity in mutants of Synechococcus PCC 7942 which are functionally deficient in an endogenous Zn2+-metallothionein gene, smtA. MT2 was also expressed as a recombinant fusion protein in Escherichia coli, purified and shown to bind Zn2+ in vitro. The mean pH of half displacement of Zn2+ from MT2 was estimated to be 5.05. This suggests that MT2 has a greater affinity for Zn2+ than phytochelatins. The results presented here reveal that MT2 is capable of binding Zn2+ in vitro, conferring tolerance to elevated [Zn2+] in vivo within cyanobacteria and is likely to compete with other polypeptides for cellular Zn2+ in planta. PMID- 8704128 TI - Isolation and characterization of two cDNA clones for mRNAs that are abundantly expressed in immature anthers of rice (Oryza sativa L.). AB - The relationship between the length of anthers and the stage of development of microspores was examined in rice (Oryza sativa L. cv. Hayayuki). Anthers of < or = 2 mm and 2.1-2.2 mm in length and those ready to dehiscence were determined to be at the uninucleate, binucleate and trinucleate microspore stage, respectively. Two cDNAs (YY1 and YY2), representing genes that are specifically expressed in anthers at the uninucleate microspore stage, were isolated and characterized. YY1 cDNA encoded an open reading frame of 95 amino acids. Eight cysteine residues with the potential to form disulfide bridges were present in the amino acid sequence. There was a hydrophobic region at the N-terminus of the putative protein, suggesting that the YY1 protein might be secreted. This cysteine motif and the hydrophobic N-terminus are conserved among products of several anther specific genes or cDNAs isolated from various plant species. These proteins are thought to form a superfamily of proteins that are confined to anthers. The YY1 transcript was localized in the tapetal cells and the peripheral cells of the vascular bundle. YY2 cDNA encoded an open reading frame of 389 amino acids and the deduced amino acid sequence exhibited substantial homology to that of chalcone synthase. Expression of YY2 mRNA was confined to the tapetal cells. The genes correspond to YY1 and YY2 cDNAs were shown to exist as single copies in the rice genome. PMID- 8704129 TI - RNA-binding proteins of 37/38 kDa bind specifically to the barley chloroplast psbA 3'-end untranslated RNA. AB - The stability of the psbA mRNA increases during barley chloroplast development eventually reaching a half-life of over 40 h. Translation of psbA mRNA is also regulated in a complex way. Sequence-specific RNA binding proteins may modulate the translation or stability of the psbA mRNA during chloroplast development. UV cross-linking assays revealed that chloroplast proteins of 37 and 38 kDA bind specifically to the 3' end of psbA transcripts and not to the 5' end of psbA or rbcL transcripts. The two RNA-binding proteins were partially purified by ammonium sulfate precipitation followed by heparin agarose chromatography. Deletion and site-directed mutation analysis demonstrated that the 37/38RNPs bind in a 30 nucleotide region immediately downstream from the translation termination codon and upstream of sequences capable of forming a stem-loop structure in the 3' end of psbA transcripts. Single-base changes that diminish the binding of the 37RNP also reduce binding of the 38RNP suggesting that these proteins may bind as a heterodimer. The 37/38RNPs that bind within the 3' end of psbA transcripts could modulate transcription termination, translation or mRNA stability. PMID- 8704130 TI - Characterization of common cis-regulatory elements responsible for the endosperm specific expression of members of the rice glutelin multigene family. AB - Glutelin is the most abundant storage protein in rice, which is expressed specifically in the endosperm of maturing seed. Glutelin is encoded by about 10 genes per haploid genome, which are clearly divided into two subfamilies (GluA and GluB). Most of them are coordinately expressed during seed maturation in spite of the remarkable divergence in the 5'-flanking regions between members of two subfamilies. In order to identify the common regulatory mechanisms responsible for the endosperm-specific expression, various cis-regulatory elements in the 5'-flanking region of the glutelin GluB-1 gene were characterized by studying the expression of chimeric genes that consisted of the sequentially deleted or mutagenized promoter and a beta-glucuronidase (GUS) reporter gene in transgenic tobacco seeds. The essential cis-regulatory elements governing the spatially and temporally specific expression of the glutelin gene expression were located within the first 245 bp of the promoter region of the GluB-1 gene from the site of initiation of transcription. The AACA motif between positions -73 and -61 common to all the six genes for glutelin sequenced to date and is repeated between positions -212 and -200 is implicated in the seed-specific expression. The GCN4 motif between positions -165 and -158 and between positions -96 and -92 that is conserved at homologous sites in all the members of glutelin gene family is also involved in the seed-specific regulation. However, both are required for the high level of seed-specific expression, because deletion of the region containing one set of both elements or substitution mutation of the AACA or GCN4 motif substantially reduced the activity. As a whole, our results suggest the combinatorial interaction of the elements in regulation of the glutelin gene expression. PMID- 8704131 TI - Evolutionary conservation and expression patterns of maize starch branching enzyme I and IIb genes suggests isoform specialization. AB - Expression of the maize (Zea mays L.) starch branching enzyme (SBE) genes Sbe1 and Sbe2 were characterized during kernel development and in vegetative tissues. The onset of Sbe1 and Sbe2 expression during endosperm development was similar to that of other genes involved in starch biosynthesis (Wx, Sh2 and Bt2). However, the expression of Sbe2 peaked earlier than that of Sbe1 in developing endosperm and embryos resulting in a shift in the ratio of Sbe1 to Sbe2 relative message levels during kernel and embryo development. Transcripts hybridizing to the Sbe2 probe were not detectable in leaves or roots which nonetheless have SBEII enzymatic activity, suggesting that there may be another divergent SBEII-like gene(s) in maize. A similar expression pattern is shared between the maize genes and related genes in pea, which together with their evolutionary conservation, suggests that the SBE isoforms may play unique roles in starch biosynthesis during plant development. PMID- 8704132 TI - Induction of cysteine and serine proteases during xylogenesis in Zinnia elegans. AB - The terminal process of xylogenesis, autolysis, is essential for the formulation of a tubular system for conduction of water and solutes throughout the whole plant. Several hydrolase types are implicated in autolysis responsible for the breakdown of cytoplasm. Here, we characterize p48h-17 cDNA from in vitro tracheary elements (TEs) of Zinnia elegans which encodes a preproprotein similar to papain. The putative mature protein, a cysteine protease, has a molecular mass of 22,699 Da with a pI of 5.7. DNA gel blot analysis indicated that p48h-17 is likely encoded by one or two genes. The p48h-17 mRNA accumulated markedly in in vitro differentiating TEs, whereas it appeared not to be induced in response to senescence and wounding in the leaves or H2O2 challenge in the cultured mesophyll cells. In stems, the expression of the p48h-17 gene was preferentially associated with differentiating xylem. Activity gel assays demonstrated that a cysteine and a serine protease, which had apparent molecular masses of 20 kDa and 60 kDa, respectively, were markedly induced during in vitro TE differentiation. The cysteine protease activity was also preferentially present in the xylem of Zinnia stems. Transient expression of the p48h-17 cDNA in tobacco protoplasts resulted in the production of a 20 kDa cysteine protease. Taken together, the results indicate that the p48h-17 gene appears to be preferentially associated with xylogenesis, and both the cysteine and serine proteases might be involved in autolysis during xylogenesis. PMID- 8704133 TI - Identification of tissue-specific, dehydration-responsive elements in the Trg-31 promoter. AB - Trg-31 is a pea gene which is rapidly induced in response to dehydration stress. Trg-31 belongs to a family of membrane intrinsic proteins that play a role in facilitating inter-membrane transport. In order to understand the mechanism of regulation of Trg-31 expression we defined the cis-acting elements of the Trg-31 promoter and the trans-acting factors that interact with them. Deletion analysis of the Trg-31 promoter in transgenic tobacco plants indicated the presence of two positive regulatory region: (1) between the -1412 to -1066 nucleotides and (2) between the -559 to -391 nucleotides, with respect to the transcription initiation site. A negative regulatory element was also identified between -1066 and -559 nucleotides. All three regulatory regions specifically regulated Trg-31 induction in leaf tissue and had little or no effect on Trg-31 induction in root tissue. Gel-mobility shift assays indicated the presence of leaf nuclear proteins that interacted with the -1412 to -1066 and -1066 to -559 regions and an 'ABRE like' sequence at -74 in the 5' promoter region of Trg-31. PMID- 8704134 TI - Expression of three members of the calcium-dependent protein kinase gene family in Arabidopsis thaliana. AB - Calcium-dependent protein kinases (CDPKs) belong to a unique family of enzymes containing a single polypeptide chain with a kinase domain at the amino terminus and a putative calcium-binding EF hands structure at the carboxyl terminus. From Arabidopsis thaliana, we have cloned three distinct cDNA sequences encoding CDPKs, which were designated as atcdpk6, atcdpk9 and atcdpk19. The full-length cDNA sequences for atcdpk6, atcdpk9 and atcdpk19 encode proteins with a molecular weight of 59343, 55376 and 59947, respectively. Recombinant atCDPK6 and atCDPK9 proteins were fully active as kinases whose activities were induced by Ca2+. Biochemical studies suggested the presence of an autoinhibitory domain in the junction between the kinase domain and the EF hands structure. Serial deletion of the four EF hands of atCDPK6 demonstrated that the integrity of the four EF hands was crucial to the Ca2+ response. All the three atcdpk genes were ubiquitiously expressed in the plant as demonstrated by RNA gel blot experiments. Comparison of the genomic sequences suggested that the three cdpk genes have evolved differently. Using antibodies against atCDPK6 and atCDPK9 for immunohistochemical experiments, CDPKs were found to be expressed in specific cell types in a temporally and developmentally regulated manner. PMID- 8704135 TI - Sugars act as signal molecules and osmotica to regulate the expression of alpha amylase genes and metabolic activities in germinating cereal grains. AB - The molecular mechanisms that initiate and control the metabolic activities of seed germination are largely unknown. Sugars may play important roles in regulating such metabolic activities in addition to providing an essential carbon source for the growth of young seedlings and maintaining turgor pressure for the expansion of tissues during germination. To test this hypothesis, we investigated the physiological role of sugars in the regulation of alpha-amylase gene expression and carbohydrate metabolism in embryo and endosperm of germinating rice seeds. RNA gel blot analysis revealed that in the embryo and aleurone cells, expression of four alpha-amylase genes was differentially regulated by sugars via mechanisms beyond the well-known hormonal control mechanism. In the aleurone cells, expression of these alpha-amylase genes was regulated by gibberellins produced in the embryo and by osmotically active sugars. In the embryo, expression of two alpha-amylase genes and production of gibberellins were transient, and were probably induced by depletion of sugars in the embryo upon imbibition, and suppressed by sugars influx from the endosperm as germination proceeded. The deferential expression of the four alpha-amylase genes in the embryo and aleurone cells was probably due to their markedly different sensitivities to changes in tissue sugar levels. Our study supports a model in which sugars regulate the expression of alpha-amylase genes in a tissue-specific manner: via a feedback control mechanism in the embryo and via an osmotic control mechanism in the aleurone cells. An interactive loop among sugars, gibberellins, and alpha-amylase genes in the germinating cereal grain is proposed. PMID- 8704136 TI - A gene for Brassica napus extensin is differentially expressed on wounding. AB - We have analysed the expression of the endogenous extensin genes in Brassica napus, using northern hybridisation and dot blotting. In the unstressed plant, the extA gene is only expressed in the root, expression in the leaf, petiole and stem being absent. We have found that wounding dramatically alters this normal pattern of expression. Expression in wounded leaf is seen after 36 h, in wounded petioles after 11 h and in wounded stem after 17 h. Differences in the amount of extensin mRNA accumulated are also seen: wounded petiole accumulating extensin message to a level higher than the leaf or the stem. Inhibitors of ethylene biosynthesis greatly delay the onset of accumulation of extensin mRNA in wounded tissues. Wounding the root causes the level of extensin message to decline with time, until levels below the limit of non-specific hybridisation are reached 11 h after wounding. Thus, application of the wounding stimulus results in the accumulation of extensin gene transcripts to different degrees and at different times in the aerial parts of the plant, and results in a decline in the same transcripts in the roots. Extensin transcript accumulation as a result of wounding is also dependent on the age of the tissue; high levels of message are seen in old wounded leaves, while expression in young wounded leaves is absent. PMID- 8704137 TI - cDNA encoding a wheat (Triticum aestivum cv. Chinese spring) glycine-rich RNA binding protein. AB - A wheat cDNA encoding a glycine-rich RNA-binding protein, whGRP-1, was isolated. WhGRP-1 contains two conserved domains, the RNA-binding motif (RNP motif) combined with a series of glycine-rich imperfect repeats, characteristic of a conserved family of plant RNA-binding proteins. Northern analysis revealed that whGRP-1 mRNA accumulates to high levels in roots and to lower levels in leaves of wheat seedlings, whGRP-1 mRNA accumulation is not enhanced by exogenous abscisic acid in seedlings and accumulates to very high levels during wheat embryo development, showing a pattern different from that of the ABA-inducible wheat Em gene. PMID- 8704139 TI - Sequence analysis of a cDNA containing the gag and prot regions of the soybean retrovirus-like element, SIRE-1. AB - SIRE-1 is a family of several hundred dispersed copies of a very large DNA element from Glycine max that has features characteristic of retroviruses and retrotransposons. A 2.4 kb SIRE-1-specific fragment was recovered from a soybean cDNA library and sequenced. The sequence contains two ORFs. Theoretical translation of ORF1 produces a gag-prot-like polyprotein containing highly conserved motifs found in retroelement nucleocapsids (CX2CX4HX4C) and aspartic proteases (LDSG). The second ORF is foreshortened. The cDNA also contains nearly 200 bp of a putative 5' LTR just upstream of a tRNA primer-binding site. PMID- 8704138 TI - Cloning and characterization of the chlorophyll biosynthesis gene chlM from Synechocystis PCC 6803 by complementation of a bacteriochlorophyll biosynthesis mutant of Rhodobacter capsulatus. AB - A bacteriochlorophyll a biosynthesis mutant of the purple photosynthetic bacterium Rhodobacter capsulatus was functionally complemented with a cosmid genomic library from Synechocystis sp. PCC 6803. The complemented R. capsulatus strain contains a defined mutation in the bchM gene that codes for Mg protoporphyrin IX methyltransferase, the enzyme which converts Mg-protoporphyrin IX to Mg-protoporphyrin IX methylester using S-adenosyl-L-methionine as a cofactor. Since chlorophyll biosynthesis also requires the same methylation reaction, the Synechocystis genome should similarly code for a Mg-protoporphyrin IX methyltransferase. Sequence analysis of the complementing Synechocystis cosmid indicates that it contains an open reading frame exhibiting 29% sequence identity to BchM. In addition, expression of the Synechocystis gene in the R. capsulatus bchM mutant via the strong R. capsulatus puc promoter was shown to support nearly wild-type levels of bacteriochlorophyll a synthesis. To our knowledge, the Synechocystis sequence thus represents the first chlorophyll biosynthesis gene homolog of bchM. The complementing Synechocystis cosmid was also shown to code for a gene product that is a member of a highly conserved family of RNA binding proteins, the function of which in cyanobacteria remains undetermined. PMID- 8704140 TI - Retrotransposon BARE-1 is a major, dispersed component of the barley (Hordeum vulgare L.) genome. AB - The barley BARE-1 is a transcribed, copia-like retroelement with well-conserved functional domains, an active promoter, and a copy number of at least 3 x 10(4). We examined its chromosomal localization by in situ hybridization. The long terminal repeat (LTR) probe displayed a uniform hybridization pattern over the whole of all chromosomes, excepting paracentromeric regions, telomeres, and nucleolar organizer (NOR) regions. The integrase probe showed a similar pattern. The 5'-untranslated leader (UTL) probe, expected to be the most rapidly evolving component, labeled chromosomes in a dispersed and non-uniform manner, concentrated in the distal regions, possibly indicating a targe site preference. PMID- 8704141 TI - The mRNA for an ETR1 homologue in tomato is constitutively expressed in vegetative and reproductive tissues. AB - Dominant mutations in the Arabidopsis ETR1 gene block the ethylene signal transduction pathway. The ETR1 gene has been cloned and sequenced. Using the ETR1 cDNA as a probe, we identified a cDNA homologue (eTAE1) from tomato. eTAE1 contains an open reading frame encoding a polypeptide of 754 amino acid residues. The nucleic acid sequence for the coding sequence in eTAE1 is 74% identical to that for ETR1, and the deduced amino acid sequence is 81% identical and 90% similar. Genomic Southern blot analysis indicates that three or more ETR1 homologues exist in tomato. RNA blots show that eTAE1 mRNA is constitutively expressed in all the tissues examined, and its accumulation in leaf abscission zones was unaffected by ethylene, silver ions (an inhibitor of ethylene action) or auxin. PMID- 8704142 TI - Expression of the Volvox gene encoding nitrate reductase: mutation-dependent activation of cryptic splice sites and intron-enhanced gene expression from a cDNA. AB - Use of the nitrate reductase encoding gene (nitA) as selection marker has facilitated the successful nuclear transformation of Volvox carteri. The Volvox nitA gene contains 10 introns. A stable nitA mutation in the Volvox recipient strain 153-81 resides in a G-to-A transition of the first nucleotide in the 5' splice site of nitA intron 2. This mutation resulted in at least three non functional splice variants, namely: (1) intron 2 was not spliced at all; (2) a cryptic 5' splice site 60 nt upstream or (3) a cryptic 5' splice site 16 nt downstream of the mutation were activated and used for splicing. When we used nitA cDNA (pVcNR13) for transformation of V. carteri 153-81, a low efficiency of about 5 x 10(-5) transformants per reproductive cell was observed. Re-integration of either intron 1 (pVcNR15) or introns 9 and 10 (pVcNR16) in the transforming cDNA increased transformation rates to 5 x 10(-4). In parallel, pVcNR15 transformed Volvox exhibited growth rates that were 100-fold increased over the pVcNR13-transformed alga. This intron-enhancement of nitA gene expression appears to be associated with post-transcriptional processing and 'channelling' of the message. These data suggest an important role of splicing for gene expression in V. carteri. PMID- 8704143 TI - Evidence for the expression of the triosephosphate translocator gene in green and non-green tissue of tomato and potato. AB - Western blot analysis revealed a cross reaction of an antibody against the spinach triosephosphate translocator with 29 kDa proteins from envelope membranes of plastids from green and red tomato fruits and also of potato tuber amyloplasts. Envelope membranes from potato tubers were isolated from a homogenate of total membranes by isopycnic sucrose density gradient centrifugation. We were able to demonstrate by reverse transcription and sequencing of the PCR product that the mRNA for the triosephosphate translocator in leaves is also present in green and red tomato fruits. The mature protein consists of 330 amino acid residues and is highly homologous to the triosephosphate translocator proteins from potato and tobacco. The PCR product obtained for potato tubers was partly sequenced. It corresponds entirely to the cDNA sequence encoding the potato leaf triosephosphate translocator protein. Evidence for the expression of the triosephosphate translocator gene in various photosynthetic active and inactive tomato tissues (leaf, green fruit, red fruit, root, petal, sepal) and potato tubers was further confirmed by northern blot analysis. PMID- 8704144 TI - Molecular analysis of the gene family of the signal recognition particle (SRP) RNA of tomato. AB - The sequence variants of the signal recognition particle (SRP) RNA gene family from four tomato cultivars have been isolated and characterized which indicated the existence of SRP RNA pseudogenes. Sequence analysis revealed two conserved sequence motifs in the upstream region, a TATA-like box and an upstream sequence element (USE), 'TCCCACATCG', both located at a conserved distance to the transcription start point. These elements are identical to the DNA-dependent RNA polymerase III (pol III)-specific promoters of U-rich small nuclear RNA (UsnRNA) genes of plants. Moreover, T-rich stretches are found at the 3' end of the coding regions of the SRP RNA genes which could act as typical pol III termination signals. These findings and recent results from site-directed mutation analysis of the SRP RNA genes from Arabidopsis thaliana indicate that, in contrast to mammalian systems, plant pol III SRP RNA genes are most probably regulated by external promoter elements. According to the identical promoter organization between plant U3-, U6snRNA, MRP-like RNA and SRP RNA genes, one can group these genes into the 'pol III(EXT)USE' subclass of externally regulated USE-dependent pol III genes. PMID- 8704146 TI - Temperature-dependent increase in the DNA-binding activity of a heat shock factor in an extract of tobacco cultured cells. AB - The DNA-binding activity of a tobacco heat shock factor (HSF) was induced by heat treatment (37-40 degrees C) of a cell-free extract that contained extra-nuclear fraction, but not in an extract of isolated nuclei. These observations suggest that an inactive form of HSF can directly recognize and transduce the heat shock signal and that such transduction requires components of the extranuclear fraction. Addition of ATP or of most other nucleoside triphosphates reduced the binding of the HSF to the heat shock element (HSE) in the same extract, and removal of ATP by dialysis from the extract restored the ability of the HSF to bind to DNA. The restored activity of the HSF could be eliminated again by a second addition of ATP. Our observations provide the first example of the involvement of ATP in the regulation of the reversible changes in HSF that control its ability to bind to HSEs in a cell-free extract. PMID- 8704145 TI - Expression of DC8 is associated with, but not dependent on embryogenesis. AB - DC8 is a late embryogenesis-abundant (LEA) protein gene isolated from carrot (Daucus carota). Deletion analysis of the DC8 promoter was performed to determine the sequences required for ABA and seed-specific regulation of DC8 transcription. To investigate the mechanism of DC8 expression during seed development, chimeric gene constructs containing DC8 promoter fragments fused to a promoterless beta glucuronidase gene (DC8:GUS) were introduced into carrot, tobacco (Nicotiana tobacum) and Arabidopsis thaliana plants. Seed-specific DC8 expression patterns was conserved among the three plant species. However, differences among the species in the patterns of DC8 expression in the embryo and endosperm that correlated with differences in the rates of embryo and endosperm growth were found. Lack of correspondence between DC8 activation and embryo development among the seeds of the three species suggests that DC8 expression, which is associated with seed maturation, is not coupled to the embryo development program. The presence of DC8 activity in carrot callus and endosperm is consistent with the notion that DC8 expression is independent of embryo morphogenesis. A similar DC8 activity time-course during callus induction and seed development suggests that explantation and 2,4-D treatment initiates a course of events similar to that in the carrot ovule. After fertilization, two pathways one leading to embryo development and another to seed maturation are initiated, but they are not closely linked. As a result we find DC8, part of the maturation program, being activated at different embryonic stages in different plant species. PMID- 8704147 TI - Molecular characterization of the mitochondrial citrate synthase gene of an acidless pummelo (Citrus maxima). AB - Pummelo (Citrus maxima [Burm.] Merrill) cDNAs encoding mitochondrial citrate synthase (mCS) were cloned by reverse transcription of juice-sac poly(A)+ mRNA, followed by Taq Polymerase-mediated amplification. The nucleotide sequence of the citrus gene (cit1) is 77% conserved relative to plant mRNAs for mCS. The encoded polypeptide includes a mitochondrial targeting signal at its amino terminus; all 20 putative active-site residues of the citrus enzyme are conserved. Southern hybridization showed that citrus cit1 is a single-copy gene. A polymorphism associated with cit1 did not cosegregate with fruit acidity indicating that acitric, the gene causing the acidless phenotype of pummelo 2240, is not an allele of cit1 locus. Quantitative detection of cit1 mRNA showed that transcript levels are not developmentally regulated in juice sacs; no differences were observed between high- and low-acid genotypes. PMID- 8704148 TI - The pea early nodulin gene PsENOD7 maps in the region of linkage group I containing sym2 and leghaemoglobin. AB - The early nodulin gene, PsENOD7, is expressed in pea root nodules induced by Rhizobium leguminosarum bv. viciae, but not in other plant organs. In situ hybridization showed that this gene is transcribed during nodule maturation in the infected cells of the proximal part of the prefixation zone II. At the transition of zone II into interzone II-III, the level of PsENOD7 mRNA drops markedly. PsENOD7 has no significant homology to other genes. RFLP mapping studies have shown that PsENOD7 is located in linkage group I between the leghaemoglobin genes and sym2. PMID- 8704149 TI - Heat shock elements are involved in heat shock promoter activation during tobacco seed maturation. AB - The soybean Gmhsp 17.3-B heat shock promoter is developmentally regulated in transgenic tobacco, as indicated by the constitutive expression of a beta glucuronidase reporter in seeds [16]. In this paper, we show that both the heat shock promoter-driven beta-glucuronidase activity and the mRNA of the endogenous Nthsp18P gene accumulate coincident with the onset of seed desiccation. Deletions of the soybean Gmhsp17.3-B promoter, encompassing the heat shock element (HSE) containing regions, revealed a co-localization of sequences responsible for heat induction and developmental expression. Moreover, synthetic HSEs fused to a TATA box sequence had the potential to stimulate the developmental expression of a GUS reporter gene in seeds of transgenic plants. PMID- 8704150 TI - Common occurrence of homologues of petunia glycine-rich protein-1 among plants. AB - The presence of specific glycine-rich proteins (GRP) related to petunia GRP1 (ptGRP1) was examined in three species of monocots (wheat, barley and maize) and five species of dicots (rape, turnip, soybean, crabapple and tomato). Protein blot analysis showed that anti-ptGRP1 antibody cross-reacted with a single different polypeptide in all species except maize. The molecular mass of these polypeptides ranged from 14 to 55 kDa. Tissue-print immunoblots of rape petioles and stems showed that the rape ptGRP1 homologue, like ptGRP1, is primarily located in the vascular tissue, and that its expression decreases with developmental age of the tissue. In barley, the ptGRP1 homologue is found in leaf vascular bundles, and may also be present in the surrounding bundle sheaths. Unlike the dicots examined, expression of the protein did not appear to decrease significantly with developmental age. PMID- 8704151 TI - Insertion of pea lectin into a phospholipid monolayer. AB - Pea lectin (PSL) is a secretory sugar-binding protein, readily soluble in aqueous solutions of low osmolarity. However, PSL also appears to be associated with the plasma membrane at the tip of young pea root hairs. By using the Wilhelmy plate method, we found that PSL can insert into a lipid monolayer. This property appeared to be independent of the sugar-binding ability of the protein. This result suggests that PSL may be directly involved in membrane-mediated interactions with saccharide ligands, for example during root hair infection by symbiotic rhizobia. PMID- 8704152 TI - Molecular characterization of an adenylate cyclase gene of the cyanobacterium Spirulina platensis. AB - A cyaA gene, encoding an adenylate cyclase, was isolated from a filamentous cyanobacterium, Spirulina platensis, by functional complementation of a cya mutant of Escherichia coli, defective in adenylate cyclase activity. The predicted gene product of cyaA contains a signal peptide-like domain, a putative sensor domain similar to the gene product of vsrA of Pseudomonas solanacearum, a putative membrane-spanning domain and an adenylate cyclase-like catalytic domain. Two other positive clones that complemented the E. coli mutant were isolated from the same cyanobacterium, suggesting that several cya genes are functioning in S. platensis. PMID- 8704153 TI - On the role of the N-terminus of the extrinsic 33 kDa protein of Photosystem II. AB - The role of the N-terminus of the extrinsic 33 kDa protein of Photosystem II has been investigated by means of site-directed mutagenesis and cross-linking. Replacement of Asp-9 resulted in a dramatic increase in proteolytic sensitivity leading to the degradation of the protein forming a 31 kDa fragment with an undefined N-terminus. This fragment was unable to restore oxygen evolution. However, the variants of the 33 kDa protein which remained intact could reconstitute oxygen evolution as effectively as the wild-type protein. Cross linking experiments with a water-soluble carbodiimide revealed that mutagenesis of residue D9 led to the disruption of an intramolecular salt bridge. Therefore we suggest that the N-terminus of the 33 kDa protein is necessary for maintaining the binding ability of the protein to Photosystem II but might not be involved in binding itself. PMID- 8704154 TI - PCR and sequencing from a single pollen grain. AB - In order to eliminate the laborious step of DNA extraction preceding all studies within the field of plant molecular biology we attempted to do PCR amplifications directly on pollen grains. Successful PCR amplification was obtained in reactions including a single pollen grain from Hordeum vulgare or Secale strictum. Both the plastid gene encoding ribulose-1,5-biphosphate carboxylase/oxygenase (rbcL) and the nuclear-encoded internal transcribed spacer regions (ITS) and the 5.8S rDNA region were amplified and sequenced to verify PCR amplification. PMID- 8704155 TI - Physical and gene maps of the unicellular cyanobacterium Synechococcus sp. strain PCC6301 genome. AB - A physical map of the unicellular cyanobacterium Synechococcus sp. strain PCC6301 genome has been constructed with restriction endonucleases PmeI, SwaI, and an intron-encoded endonuclease I-CeuI. The estimated size of the genome is 2.7 Mb. On the genome 49 genes or operons have been mapped. Two rRNA operons are separated by 600 kb and transcribed oppositely. PMID- 8704156 TI - Isolation of a full-length cDNA encoding calreticulin from a PCR library of in vitro zygotes of maize. AB - A full-size cDNA clone (1614 bp) encoding calreticulin was isolated from a PCR based cDNA library of maize in vitro zygotes. Calreticulin is a major Ca2+ storage protein located mainly in the lumen of the endoplasmic reticulum but also in the nucleus and/or cytoplasm of some cells. A differential screening between cDNA libraries originating from 104 in vitro zygotes (18 h after in vitro fertilization) and 128 unfertilized egg cells was performed to isolate newly expressed genes or genes expressed more abundantly after fertilization. The expression of the isolated cDNA clone is enhanced after fertilization and strongly correlated to cell division. Sequence comparison to a shorter maize calreticulin cDNA isolated from a PCR based cDNA library construction from a few plant cells [12]. It is further shown that calreticulins in maize are probably transcribed from a small gene family differentially expressed in abundance in diverse tissues. The deduced amino acid sequence encodes an acidic protein (pI 4.17) of 48 kDa sharing 77-92% and 50-54% homology to other plant and animal calreticulins, respectively. The described calreticulin gene represents to our knowledge the first cDNA clone isolated from a RT/PCR cDNA library originating from only a few plant cells and is the first gene isolated from zygotes of higher plants. PMID- 8704157 TI - A vicilin-like seed protein of cycads: similarity to sucrose-binding proteins. AB - Seed storage globulins of the 7S and 11S type are synthesized in the seeds of angiosperms and gymnosperms. We have isolated and characterized a vicilin-like gene expressed in the cycad Zamia furfuraceae. Sequence comparisons reveal clear similarities to a sucrose-binding protein isolated from soybean. We suggest the existence of a superfamily of related genes including both vicilin-like and legumin-like seed globulin genes as well as genes coding for spherulins, germins and sucrose-binding-proteins. PMID- 8704158 TI - Regulation of cytosolic pyruvate, orthophosphate dikinase expression in developing maize endosperm. AB - Pyruvate orthophosphate dikinase (PPDK, E.C. 2.7.9.1) is an abundant enzyme in the leaves of C4 plants associated with the dicarboxylic acid pathways of CO2 fixation in the dark. PPDK activity has also been detected in the seeds of maize and other, non-C4 cereals, where its role has yet to be established. Using an anti-PPDK serum, two cross-reacting species of M(r) close to 90 000 were detected in developing maize endosperm of wild-type plants. In two independent opaque-2 mutant lines, one of the polypeptides was absent and the other was reduced in level. Similarly, endosperm PPDK mRNA levels were greatly reduced in the opaque-2 maize lines compared to wild type, suggesting that endosperm PPDK gene expression is under Opaque-2 control. However, a low level of PPDK mRNA could still be detected in these mutants, indicating that PPDK gene expression is not absolutely dependent on Opaque-2 but rather can be modulated by it. This interpretation was reinforced by the demonstration that the distribution of PPDK transcripts is not affected in o2 mutants, although the level is reduced, and that PPDK mRNA is detectable prior to 02 mRNA during the maturation of wild-type maize endosperm. Using oligonucleotides specific for the different maize PPDK genes, the o2 mutations were shown to affect only cyPPDKZml gene expression in maize line A69Y. PMID- 8704159 TI - Isolation of a novel RNA-binding protein and its association with a large ribonucleoprotein particle present in the nucleoplasm of tobacco cells. AB - A cDNA encoding a protein with a consensus sequence-type RNA-binding domain (CS RBD) has been isolated from a Nicotiana sylvestris cDNA library. The deduced protein (designated 'RZ-1') contains CS-RBD in its N-terminal half, arginine/aspartic acid repeats in its center and a glycine-rich-C-terminal region in which a zinc finger motif of the CCHC type is present. The corresponding gene appears to be expressed constitutively in all tobacco organs. Immunocytochemical assays revealed that RZ-1 is localized in the nucleoplasm of tobacco cultured cells. Glycerol gradient fractionation of tobacco nuclear lysates showed that RZ 1 is associated with a large ribonucleoprotein particle of around 60 S in size. Nucleic acid-binding assays indicated that RZ-1 binds preferentially to poly (G) and both the CS-RBD and glycine-rich region are necessary for its binding activity. A possible role of RZ-1 is discussed. PMID- 8704160 TI - Cloning of a cDNA encoding a 3-dehydroquinate synthase from a higher plant, and analysis of the organ-specific and elicitor-induced expression of the corresponding gene. AB - cDNA clones for all enzymes of the prechorismate pathway of higher plants have previously been cloned, with the exception of the second enzyme of the pathway, 3 dehydroquinate synthase. Here we describe the isolation of a cDNA encoding a 3 dehydroquinate synthase from tomato which was identified by complementing a 3 dehydroquinate synthase-deficient Escherichia coli strain with a tomato cDNA library. The deduced amino acid sequence contains a putative N-terminal plastid specific transit peptide, and the sequence of the mature enzyme resembles those of the corresponding bacterial enzymes more than of the fungal enzymes. Sequence identity was even higher between the tomato and E. coli sequences than between the E. coli and other known bacterial sequences. The abundance of 3 dehydroquinate synthase transcripts differ in the organs of tomato plants analyzed. In cultured tomato cells, the abundance of 3-dehydroquinate synthase transcripts increased 9-fold within 4 to 5 h of elicitor treatment. PMID- 8704161 TI - Elongation factor 1 alpha genes of the red alga Porphyra purpurea include a novel, developmentally specialized variant. AB - The life cycle of the red alga Porphyra purpurea alternates between two morphologically distinct phases: a shell-boring, filamentous sporophyte and a free-living, foliose gametophyte. From a subtracted cDNA library enriched for sporophyte-specific sequences, we isolated a cDNA encoding an unusual elongation factor 1 alpha (EF-1 alpha) that is expressed only in the sporophyte. A second EF 1 alpha gene that is expressed equally in the sporophyte and the gametophyte was isolated from a genomic library. These are the only EF-1 alpha genes detectable in P. purpurea. The constitutively expressed gene encodes an EF-1 alpha very similar to those of most eukaryotes. However, the sporophyte-specific EF-1 alpha is one of the most divergent yet described, with nine insertions or deletions ranging in size from 1 to 26 amino acids. This is the first report of a developmental stage-specific EF-1 alpha outside of the animal kingdom and suggests a fundamental role for EF-1 alpha in the developmental process. PMID- 8704163 TI - The ITP practice guideline: what, why, and for whom? PMID- 8704162 TI - A novel kinesin-like protein with a calmodulin-binding domain. AB - Calcium regulates diverse developmental processes in plants through the action of calmodulin. A cDNA expression library from developing anthers of tobacco was screened with 35S-labeled calmodulin to isolate cDNAs encoding calmodulin-binding proteins. Among several clones isolated, a kinesin-like gene (TCK1) that encodes a calmodulin-binding kinesin-like protein was obtained. The TCK1 cDNA encodes a protein with 1265 amino acid residues. Its structural features are very similar to those of known kinesin heavy chains and kinesin-like proteins from plants and animals, with one distinct exception. Unlike other known kinesin-like proteins, TCK1 contains a calmodulin-binding domain which distinguishes it from all other known kinesin genes. Escherichia coli-expressed TCK1 binds calmodulin in a Ca(2+) dependent manner. In addition to the presence of a calmodulin-binding domain at the carboxyl terminal, it also has a leucine zipper motif in the stalk region. The amino acid sequence at the carboxyl terminal of TCK1 has striking homology with the mechanochemical motor domain of kinesins. The motor domain has ATPase activity that is stimulated by microtubules. Southern blot analysis revealed that TCK1 is coded by a single gene. Expression studies indicated that TCK1 is expressed in all of the tissues tested. Its expression is highest in the stigma and anther, especially during the early stages of anther development. Our results suggest that Ca2+/calmodulin may play an important role in the function of this microtubule-associated motor protein and may be involved in the regulation of microtubule-based intracellular transport. PMID- 8704164 TI - A cell surface marker gene transferred with a retroviral vector into CD34+ cord blood cells is expressed by their T-cell progeny in the SCID-hu thymus. AB - Gene transduction into immature hematopoietic cells collected at birth from the umbilical cord could be useful for the treatment of genetic or acquired disorders of the hematopoietic system diagnosed during pregnancy. The SCID-hu mouse is a convenient model to investigate T-cell lineage gene therapy, since it allows replication of human intrathymic T-cell development. CD34+ cells isolated from cord blood were cocultured with CRIP MFG-murine CD2 (mCD2) cells that produce recombinant retroviruses encoding the mCD2 antigen, a cell surface marker easily detectable by flow cytometry. After 3 and 4 days in coculture, a mean of 19% and 39% human hematopoietic cells, respectively, expressed the mCD2 antigen. CD34+ cells cocultured for 4 days were used to reconstitute human fetal thymus implanted in SCID mice. Five to 10 weeks later, the mCD2 antigen was detected on approximately 10% of human thymocytes repopulating the thymic grafts in four of nine SCID mouse chimeras. Vector genomes were detected in graft cell DNA by Southern blot. Analysis of vector integration indicated that positive cells were of polyclonal origin in three animals and predominantly monoclonal in the other one. Our data show that foreign genes can be transduced into CD34+ cord blood cells endowed with T-cell differentiation potential, and suggest strategies for T cell lineage gene therapy in the neonate. PMID- 8704166 TI - Cross-linking CD7 on myeloblasts results in granulocyte-macrophage colony stimulating factor production. AB - CD7+CD34+ lymphohematopoietic progenitor cells in bone marrow are capable of differentiating into either lymphocytes or myeloid cells. The mechanism whereby these bipotent progenitor cells are regulated is not yet clear. In this study, we investigated the role CD7 may play in the development of bipotent cells using two myeloid progenitor cell lines, KG-1 and KG-1a, as models for such cells. Our data showed that cross-linking CD7 on KG-1 and KG-1a cells induced transcription, translation, and secretion of granulocyte-macrophage colony-stimulating factor (GM-CSF). Anti-CD7 antibody also augmented the colony formation by KG-1 cells. Protein synthesis in KG-1 cells also increased as a result of anti-CD7 stimulation. These phenomena could be blocked by anti-GM-CSF, and supported the notion that the secreted GM-CSF was the primary mediator of CD7 effects. Together, these findings suggest that the interaction between CD7 and its putative ligand may play an important role in hematopoietic development. PMID- 8704165 TI - Retinoic acid activates interferon regulatory factor-1 gene expression in myeloid cells. AB - All-trans-retinoic acid (ATRA) is the drug of choice in the treatment of acute promyelocytic leukemia (APL). ATRA induces both in vitro and in vivo differentiation of APL cells into mature granulocytes. However, the molecular mechanisms involved in ATRA-dependent growth inhibition and cellular differentiation are not presently understood. The NB4 cell line, which is derived from the bone marrow of a patient with APL during relapse, can be used as a model system to study the growth and differentiation of APL cells. Because interferon (IFN) regulatory factors (IRF-1 and IRF-2) and other IFN-inducible gene products regulate cell growth, we analyzed the effects of ATRA on the expression of these genes. We show that ATRA directly activates IRF-1 gene expression, followed by activation of IRF-2 and 2'-5' oligoadenylate synthetase (OAS) gene expression with slower kinetics. In addition to NB4 cells, ATRA also activated IRF-1 gene expression in HL-60, U937, and THP-1 cells, which all respond to ATRA by growth inhibition. A more than additive increase in IRF-1 gene expression was seen with ATRA and IFN-gamma in NB4 cells. ATRA did not activate nuclear factor kappa B or signal transducer and activator of transcription (STAT) activation pathways, suggesting that an alternate mechanism is involved in IRF-1 gene activation. The ATRA-induced expression of IRF-1, an activator of transcription and repressor of transformation, may be one of the molecular mechanisms of ATRA-induced growth inhibition, and the basis for the synergistic actions of ATRA and IFNs in myeloid leukemia cells. PMID- 8704167 TI - Exposure of human CD34+ cells to human immunodeficiency virus type 1 does not influence their expansion and proliferation of hematopoietic progenitors in vitro. AB - The susceptibility of highly purified human CD34+ cells to monocytotropic (Ba-L) and lymphotropic (A018-post) strains of human immunodeficiency virus-1 (HIV-1) was examined. Liquid cultures initiated with fresh immunomagnetically purified CD34+ cells using the K6.1 CD34 monoclonal antibody (MoAb) (K6.1/CD34+) were positive for HIV expression 2 weeks after exposure to HIV-1 Ba-L. These cells were initially greater than 90% CD34+ and had undetectable monocyte contamination by flow-cytometric staining and side-scatter analyses, respectively, and undetectable T-cell contamination by CD3 polymerase chain reaction (PCR) analysis. However, secondary CD34+ liquid cultures reselected from the primary liquid cultures 24 hours after HIV exposure by panning with the ICH3 CD34 MoAb (ICH3/CD34+) and maintained for an additional 14 days were negative for HIV expression. The ICH3-unbound cells were positive for both spliced and unspliced HIV RNA when exposed to HIV-1 Ba-L, and were DNA PCR positive when exposed to either monocytotropic or lymphotropic HIV-1. To further test that CD34+ cells were not infectible by HIV-1, we exposed K6.1/CD34+ cells continuously to HIV-1 in a culture system capable of maintaining and expanding primitive CD34+ cells. HIV-exposed K6.1/CD34+ cells proliferated and expanded as efficiently as uninfected cultures. However, when reselected magnetically using the K6.1 CD34 MoAb after expansion for 7 days, bound K6.1/CD34+ cells were again negative for HIV-1 expression, whereas unbound cells were positive for HIV-1 expression. These findings suggest that a sequential CD34+ cell-selection process, in which the two selections are separated by a brief culture period, can yield a population of CD34+ cells that are not infected with HIV-1. This process may be useful in the design of stem or progenitor cell-based transplantation therapies for HIV infection. PMID- 8704168 TI - Steel factor induces serine phosphorylation of Stat3 in human growth factor dependent myeloid cell lines. AB - Steel factor (SLF) acts synergistically with various hematopoietic growth factors that use the Jak-Stat pathways in vivo and in vitro, although the contribution by SLF to this pathway is unknown. We show here that SLF induces time- and dose dependent phosphorylation of Stat3 in the human growth factor-dependent cell lines MO7e and TF-1. This phosphorylation occurs exclusively on serine residues. Simultaneous stimulation with SLF plus other cytokines that induce tyrosine phosphorylation of Stat3, such as interleukin-9 (IL-9) in MO7e cells or IL-6 in TF-1 cells, resulted in tyrosine phosphorylation and enhanced serine phosphorylation of Stat3. Serine phosphorylation alone did not promote nuclear translocation or DNA binding activity to the sis-inducible element of Stat3. However, costimulation with SLF plus IL-9 in MO7e cells resulted in the nuclear translocation of serine-hyperphosphorylated Stat3. Serine phosphorylation of Stat3 was also observed by the stimulation of cells with granulocyte-macrophage colony-stimulating factor and IL-3, which do not induce tyrosine phosphorylation of Stat3. These results suggest that SLF might modulate the Jak-Stat3 pathway by serine phosphorylation and that the Jak-Stat pathway may be differentially regulated by the combinational stimulation of two or more cytokines. PMID- 8704169 TI - Neutrophil rolling, arrest, and transmigration across activated, surface-adherent platelets via sequential action of P-selectin and the beta 2-integrin CD11b/CD18. AB - Platelets bound to thrombogenic surfaces have been shown to support activation dependent firm adhesion of neutrophils in flow following selectin-mediated tethering and rolling. The specific receptor(s) responsible for mediating adhesion-strengthening interactions between neutrophils and platelets has not previously been identified. Furthermore, the ability of adherent platelets to support the migration of bound neutrophils has not been tested. We studied neutrophil interactions with activated, surface-adherent platelets as a model for leukocyte binding in vascular shear flow and emigration at thrombogenic sites. Our results demonstrate that the beta 2-integrin Mac-1 (CD11b/CD18) is required for both firm attachment to and transmigration of neutrophils across surface adherent platelets. In flow assays, neutrophils from patients with leukocyte adhesion deficiency-1 (LAD-I), which lack beta 2-integrin receptors, formed P selectin-mediated rolling interactions, but were unable to develop firm adhesion to activated platelets, in contrast to healthy neutrophils, which developed firm adhesion within 5 to 30 seconds after initiation of rolling. Furthermore, the adhesion-strengthening interaction observed for healthy neutrophils could be specifically inhibited by monoclonal antibodies (mAbs) to Mac-1, but not to lymphocyte function-associated antigen-1 (LFA-1; CD11a/CD18) or intercellular adhesion molecule-2 (ICAM-2; CD102). Further evidence for a beta 2-integrin dependent neutrophil/platelet interaction is demonstrated by the complete inhibition of interleukin (IL)-8-induced neutrophil transmigration across platelets bound to fibronectin-coated polycarbonate filters by mAbs to Mac-1. Thus, Mac-1 is required for firm adhesion of neutrophils to activated, adherent platelets and may play an important role in promoting neutrophil accumulation on and migration across platelets deposited at sites of vascular injury. PMID- 8704170 TI - Changes in cell spreading and cytoskeletal organization are induced by adhesion to a fibronectin-fibrin matrix. AB - Plasma fibronectin (pFN) cross-linked to fibrin during the injury response provides a provisional matrix required for cells to begin tissue repair. Using a synthetic matrix of pFN and fibrin as a substrate for cell adhesion and spreading, we have determined that pFN covalently cross-linked to fibrin into a complex multimer is functionally distinct from pFN immobilized onto a plastic surface. NIH-3T3 cells on a FN-fibrin matrix reach 50% of the maximal cell area of cells spread on FN-coated plastic. They neither attach nor spread on cross linked fibrin alone. Cells on pFN-fibrin matrices form few prominent stress fibers and exhibit clear differences in membrane ruffling and filopodial extension when stained with rhodamine-labeled phalloidin. Interestingly, these differences are enhanced by upregulation of protein kinase C. These data suggest that cell-FN interactions can be modified by the molecular context of the protein within the extracellular matrix resulting in distinct cell morphology and cytoskeletal organization. PMID- 8704171 TI - Glanzmann thrombasthenia due to a two amino acid deletion in the fourth calcium binding domain of alpha IIb: demonstration of the importance of calcium-binding domains in the conformation of alpha IIb beta 3. AB - The integrin alpha IIb beta 3, a calcium-dependent heterodimer, plays a critical role in platelet aggregation. The alpha IIb subunit of the heterodimer contains four highly conserved putative calcium-binding domains in its extracellular portion. During studies of the molecular basis of Glanzmann thrombasthenia in a child of mixed Caucasian background whose platelets expressed little alpha IIb beta 3 on their surface, we found the patient heterozygous for a two amino acid deletion in the fourth alpha IIb calcium-binding domain. When this alpha IIb mutant was expressed in COS-1 cells, we found that the deletion did not interfere with the assembly of alpha IIb beta 3 heterodimers, but altered their conformation such that they were neither recognized by the heterodimer-specific antibody A2A9 nor able to undergo further intracellular processing or transport to the cell surface. These results suggest that the calcium-binding domains in alpha IIb play an important role maintaining the overall conformation of alpha IIb beta 3. To confirm this suggestion, we deleted each of the four 12 amino acid calcium-binding domains in alpha IIb by in vitro mutagenesis and expressed the mutants along with beta 3 in COS-1 cells. Each construct formed a heterodimer with beta 3, but none of the heterodimers interacted with A2A9 or underwent further intracellular processing. These data indicate that the calcium-binding domains in alpha IIb are not involved in alpha IIb beta 3 heterodimer formation, but their presence is required for the intracellular transport of alpha IIb beta 3 to the cell surface. PMID- 8704173 TI - High doses of immunoglobulin G attenuate immune aggregate-mediated complement activation by enhancing physiologic cleavage of C3b in C3bn-IgG complexes. AB - Intravenously applied human IgG has beneficial effects in treating inflammatory diseases, presumably because it has a complement attenuating role. This role of IgG was studied in vitro by following C3 activation and inactivation in sera that were supplemented with exogenous human IgG and incubated with immune aggregates. IgG added at 2 to 10 mg/mL stimulated the physiologic inactivation of C3b containing complexes twofold to threefold in 20% sera. This, in turn, lowered the overall C3 activation by 28%, as new C3 convertases primarily assembled on C3b containing complexes. Exogenous IgG (5 mg/mL) also stimulated inactivation of purified C3b2-IgG complexes, whereby their half-life dropped from 3-4 to 1.5 minutes in 20% serum. IgG appeared to act like a modulator of factor H and I because it did not stimulate inactivation of C3b-containing complexes in factor I deficient serum. Thus, the known partial protection of C3bn-IgG complexes from inactivation by factor H and I was downregulated by high concentrations of IgG. The ability of high doses of IgG to stimulate complement inactivation is a novel regulatory role of IgG. This may be one of the molecular principles for its therapeutic efficacy in treating complement-mediated inflammations. PMID- 8704172 TI - Effects of verocytotoxin-1 on nonadherent human monocytes: binding characteristics, protein synthesis, and induction of cytokine release. AB - The epidemic form of the hemolytic uremic syndrome (HUS) has been associated with a verocytotoxin producing Escherichia coli infection. Endothelial cell damage of glomeruli and arterioles of the kidney plays a central role in the pathogenesis of HUS. A number of observations in vivo and in vitro indicate that inflammatory mediators contribute to this process. In this study we investigated the binding of 125I-verocytotoxin-1 (VT-1) to freshly isolated human nonadherent monocytes as well as the nature of the ligand to which VT-1 binds on monocytes. On the average, freshly isolated monocytes have 0.07 x 10(5) specific binding sites for 125I-VT-1 per cell. Preincubation of nonadherent monocytes with bacterial lipopolysaccharide (LPS) caused a 23- to 30-fold increase of specific binding sites for VT-1 as shown by Scatchard plot analysis. Thin-layer chromatography of extracted neutral glycolipids of the cells and subsequent binding of 125I-VT-1 showed that human monocytes bind VT-1 to a globotriaosylceramide (Gb3) species that is different from that found on endothelial cells, probably a short-chain fatty acyl Gb3 or an alpha-OH-Gb3. In addition, we evaluated the functional consequences of VT-1 binding to human monocytes by investigating the effects of VT-1 on the total protein synthesis and, specifically, the production of the cytokines interleukin-1 beta (IL-1 beta), tumor necrosis factor-alpha (TNF alpha), IL-6, and IL-8. We observed that VT-1 did not inhibit overall protein synthesis, nor under basal conditions, neither after stimulation with LPS, in contrast to previous observations with endothelial cells. Furthermore, we found that VT-1 induces the synthesis of the cytokines IL-1 beta, TNF-alpha, IL-6, and IL-8 in nonstimulated monocytes by a LPS-independent cell activation. The increase in the production of cytokines was parallelled by an increase in mRNA, as was demonstrated for IL-6 by reverse transcription-polymerase chain reaction. These data suggest that inflammatory mediators locally produced by VT-1 stimulated monocytes may contribute to the pathogenic mechanism of the HUS. PMID- 8704174 TI - Prospective evaluation of the clinical usefulness of an antigen-specific assay (MAIPA) in idiopathic thrombocytopenic purpura and other immune thrombocytopenias. AB - The diagnosis of idiopathic immune thrombocytopenia remains a clinical diagnosis based on the exclusion of other causes of immune and nonimmune thrombocytopenia. Measurement of platelet-associated Ig (PAIg), while sensitive, is nonspecific for the diagnosis of immune thrombocytopenia. Published experience of antigen capture assays (including monoclonal antibody immobilization of platelet antigens or MAIPA) suggest a high sensitivity and specificity (70% to 80%) in selected groups of patients. In a prospective evaluation of 158 patients with thrombocytopenia from all causes, we report a sensitivity of 51% and specificity of 80% for direct MAIPA assays. MAIPA was considerably better in discriminating immune from nonimmune thrombocytopenia than two assays of PAIgG. Antiplatelet antibodies detected by MAIPA were more frequently directed against the glycoprotein (GP) IIb/IIIa than the GP Ib/IX complex. Our experience suggests that MAIPA assays are useful in the laboratory assessment of thrombocytopenia, should be performed before therapy, and that some patients with 'nonimmune' thrombocytopenia may have genuine antiplatelet antibodies. PMID- 8704176 TI - Two pathways can activate the interleukin-5 gene and induce binding to the conserved lymphokine element 0. AB - Eosinophilia is a uniquely specific phenomenon regulated by interleukin-5 (IL-5), suggesting specific control for IL-5 gene expression. Using a transient transfection reporter assay and DNA mobility-shift experiments in EL4 mouse lymphoma cells, reporter expression and binding of transcription factors to the conserved lymphokine element 0 (CLE0) in the mouse (mIL-5) promoter was investigated. Activation of the IL-5 promoter required costimulation of T cells with phorbol ester (phorbol 12-myristate 13-acetate [PMA]) and cyclic adenosine 3',5'-monophosphate (cAMP), but was blocked by the immunosuppressive drug, cyclosporin A (CsA). Binding to CLE0 was induced under conditions optimal for IL 5 transcription but was not blocked by CsA. CD28-induced signals could partly substitute for cAMP. However, the effects of cAMP, but not of CD28, were sensitive to the cAMP-dependent protein kinase inhibitor, H89, suggesting that CD28 does not involve a cAMP mechanism. It therefore appears that IL-5 expression can be induced by at least two distinct stimulatory pathways. Although CLE0 contains sequences similar to AP-1 and NF-AT, only the AP-1 moiety of the CLE0 element could be demonstrated to have inducible binding. Experiments with antisera to the AP-1 family of transcription factors indicated that c-fos and JunB bind to the IL-5 CLE0 in activated lymphoma cells. The role of the NF-AT like element was less clear. A constitutively expressed protein showed a weak band that was inhibited by mIL-2 NF-AT competitor sequences. However, this protein did not react with an anti-NF-ATp antiserum. On the other hand, transcription was partially inhibited by an oligonucleotide containing the intact NF-AT-like element from CLE0, suggesting that the element is important for optimal transcription, but the nature of the protein binding to it remains unknown. The fact that these factors are induced in a subclone of EL4 that does not express IL-5 and bind to a number of other cytokine gene promoters suggests that although binding to CLE0 appears to be necessary for IL-5 transcription, other factors must control the specific expression of the gene. PMID- 8704175 TI - Granulocyte-macrophage colony-stimulating factor: an effective adjuvant for protein and peptide-based vaccines. AB - The current studies evaluate granulocyte-macrophage colony-stimulating factor (GM CSF) as a vaccine adjuvant. An important issue for developing vaccine therapy for human malignancy is identifying adjuvants that can elicit T-cell responses to proteins and peptides derived from "self" tumor antigens. GM-CSF, in vitro, stimulates the growth of antigen-presenting cells such as dendritic cells and macrophages. Initial experiments examined whether GM-CSF injected into the skin of rats could affect the number or character of antigen presenting cells, measured as class II major histocompatability complex expressing cells, in lymph nodes draining the injection site. Intradermal (id) inoculation of GM-CSF every 24 hours for a total of five inoculations resulted in an increase of class II+ fluorescing cells that peaked at the fourth inoculation. Subcutaneous (sq) inoculation resulted in an increase of class II+ fluorescing cells that peaked following the second inoculation, then decreased over time. Using this schema for "conditioning" the inoculation site, GM-CSF was administered id or sq for five injections and a foreign antigen, tetanus toxoid (tt), was given at the beginning or the end of the immunization cycle. Id immunization was more effective than sq at eliciting tt specific immunity. In addition, GM-CSF id, administered as a single dose with antigen, compared favorably with complete Freund's adjuvant (CFA) and alum in eliciting tt specific antibody and cellular immunity. We have shown that immunity to rat neu (c-erbB-2) protein, an oncogenic self protein, can be generated in rats by immunization with peptides derived from the normal rat neu sequence plus CFA. The current study demonstrates that rat neu peptides inoculated with GM-CSF could elicit a strong delayed type hypersensitivity reaction (DTH) response, whereas peptides alone were non-immunogenic. GM-CSF was as effective as CFA in generating rat neu specific DTH responses after immunization with a neu peptide based vaccine. Soluble GM-CSF is a potent adjuvant for the generation of immune responses to foreign proteins as well as peptides derived from a self tumor antigen. PMID- 8704177 TI - Telomerase activation in normal B lymphocytes and non-Hodgkin's lymphomas. AB - Activation of telomerase seems to be a prerequisite for immortalization and is found in permanent cell lines and most malignant tumors. Normal somatic cells are generally telomerase negative, except for bone marrow stem cells. Weak activity is also present in peripheral blood cells. In the present study strong telomerase activity was demonstrated in vivo in normal mature cells of the immune system, as well as in malignant lymphomas. Benign lymph nodes had lower telomerase activity than benign tonsils, which exhibited intermediate to high activity comparable with findings in malignant lymphomas. In benign tonsils the activity seemed to be restricted to germinal center B cells. In benign lymphoid tissues telomerase activity correlated with B-cell numbers and cell proliferation, but this was not observed in the lymphoma group. High-grade lymphomas exhibited higher levels of telomerase compared with low-grade cases. The data showed that in vivo activation of telomerase is a characteristic feature of germinal center B cells. Different signals for activation of telomerase are likely to exist, one of them being immune stimulation. The data suggest that telomerase activity in malignant lymphomas can be explained by an "induction and retention" model, ie, transformation occurs in a normal, mature B cell with reactivated telomerase, which is retained in the neoplastic clone. PMID- 8704178 TI - Activation of naive and memory T cells by interleukin-15. AB - Interleukin-15 (IL-15), a product of monocytes and other cells, has biological activities similar to those of IL-2, including growth stimulation of activated T cells, induction of cytolytic effector cells, and B-cell costimulation for proliferation and lg production. We report that IL-15 at optimal concentrations rapidly induced memory (CD45RO+) CD4+ and CD8+ T cells and naive (CD45RO-) CD8+ T cells to express the CD69 activation marker followed by proliferation. By contrast, IL-15 failed to induce naive (CD45RO-) CD4+ T cells to express CD69 or to proliferate. Similar findings were obtained with IL-2. Unlike the other T-cell subsets, CD4+ T cells with a naive phenotype expressed little or no IL-2R beta chain, a shared component of the IL-2 and IL-15 receptors required for receptor function. A monoclonal antibody to the IL-2R beta chain, Mik beta 1, reduced CD69 expression and proliferation in CD4+ memory, CD8+ memory, and CD8+ naive T cells activated by IL-15. These results confirm the biological similarities of IL-2 and IL-15. They further document that the pool of naive CD4+ cells, unlike the pool of memory CD4+, memory CD8+, and naive CD8+ cells, is not regulated directly by the T-cell growth factors IL-2 or IL-15. PMID- 8704179 TI - T cells from patients with Hodgkin's disease have a defective T-cell receptor zeta chain expression that is reversible by T-cell stimulation with CD3 and CD28. AB - To investigate the mechanisms underlying the deficiency of T lymphocytes from patients with Hodgkin's disease, we investigated the expression of the T-cell receptor (TCR) zeta chain in patients with Hodgkin's disease. By flow cytometry using an anti-zeta chain monoclonal antibody, peripheral blood T lymphocytes from patients with untreated Hodgkin's disease were shown to express decreased levels of the TCR zeta chain. After stimulation by combined CD3 and CD28 cross-linking, T cells from Hodgkin's disease patients upregulated zeta chain protein expression to normal values within 48 hours and achieved a cytolytic potential and levels of interleukin (IL)-2 secretion that were not different from T cells obtained from healthy controls. These results show that downregulation of the TCR zeta chain in Hodgkin's T lymphocytes is a reversible event. Costimulation of CD3 and CD28 is a novel approach for overcoming the T-cell deficiency in Hodgkin's disease and might be exploited clinically. As upregulation of the zeta chain can also be achieved using bispecific monoclonal antibodies (BI-MoAbs) with specificity for tumor antigens and CD3 and CD28, respectively, an immunotherapy with CD3/CD30 and CD28/CD30 Bi-MoAbs may overcome and should therefore, not be jeopardized by the inherent T-cell deficiency in patients with Hodgkin's disease. PMID- 8704180 TI - Epstein-Barr virus latent membrane protein-1 oncogene deletions: correlations with malignancy in Epstein-Barr virus--associated lymphoproliferative disorders and malignant lymphomas. AB - LMP-1, an Epstein-Barr viral (EBV) latency protein, is considered a viral oncogene because of its ability to transform rodent fibroblasts in vivo and render them tumorigenic in nude mice. In human B cells, EBV LMP-1 induces DNA synthesis and abrogates apoptosis. LMP-1 is expressed in EBV-transformed lymphoblastoid cell lines, nasopharyngeal carcinoma (NPC), a subset of Hodgkin's disease (HD), and in EBV-associated lymphoproliferative disorders (EBV-LPDs). Recently, focused deletions near the 3' end of the LMP-1 gene (del-LMP-1, amino acids 346-355), in a region functionally related to the half-life to the LMP-1 protein, have been reported frequently in human immunodeficiency virus (HIV) associated HD (100%) and EBV+ Malaysian and Danish peripheral T-cell lymphomas (100%, 61% respectively), but less frequently in cases of HD not associated with HIV (28%, 33%) and infectious mononucleosis (33%). To further investigate the potential relationship of del-LMP-1 to EBV-LPDs associated with immunosuppression or immunodeficiency, we studied 39 EBV-associated lymphoproliferations (10 benign, 29 malignant) from four distinct clinical settings: posttransplant (4 malignant, 1 reactive); HIV+ (18 malignant, 2 reactive); nonimmunodeficiency malignant lymphoma (ML) (7 cases); and sporadic EBV infection with lymphoid hyperplasia (7 cases). The presence of EBV within lymphoid cells was confirmed by EBV EBER1 RNA in situ hybridization or by polymerase chain reaction (PCR) analysis. EBV strain type and LMP-1 deletion status were determined by PCR. EBV strain types segregated into two distinct distributions: HIV+ (9 A; 11 B) and non HIV (19 A, 0 B), consistent with previous reports. Overall, del-LMP-1 were found in 1 of 5 (20%) Burkitt lymphomas (BL); 17 of 24 (71%) aggressive non-Hodgkin's lymphoma (agg-NHL), and 2 of 10 (20%) reactive lymphoid proliferations. Of the agg-NHLs, del-LMP-1 were present in 4 of 4 PT-ML (100%); 10 of 15 HIV+ ML (67%); and 3 of 5 nonimmunodeficiency malignant lymphoma (ML, 60%). A total of 2 of 7 (28%) sporadic EBV-associated lymphoid hyperplasias contained a del-LMP-1. All del-LMP-1 were identical by DNA sequence analysis. No correlation was identified between the presence of del-LMP-1 and the EBV strain type observed. The high incidence of del-LMP-1 observed in agg-NHLs (71%), in contrast to the relatively low incidence observed in reactive lymphoid proliferations (28%), suggests that the deleted form may be preferentially selected in lymphomatous processes. All posttransplant agg-NHLs contained a del-LMP-1, and a similar frequency of del-LMP 1 was observed in both HIV-associated ML (66%) and nonimmunodeficiency ML (60%), suggesting that impairment of immune function alone is not a requirement for the expansion of malignant cells infected by EBV stains containing the deleted LMP-1 gene. PMID- 8704181 TI - Favored use of immunoglobulin V(H)4 Genes in AIDS-associated B-cell lymphoma. AB - We examined the lg heavy chain variable region genes (Ig V(H) genes) expressed in biopsy specimens of 10 patients with acquired immunodeficiency syndrome (AIDS) associated lymphoma. Eight expressed Ig V(H) genes of the V(H)4 group, indicating a bias toward expression of Ig V(H) genes of this subgroup. Sequence analyses of Ig V(H) genes isolated from any one lymphoma did not reveal evidence for intraclonal diversity. However, some lymphomas express Ig V(H) genes that apparently have undergone somatic diversification and selection. In addition, we found that the sequence encoding each examined third complementarity determining region most likely resulted from D-D fusion, a process that ordinarily contributes to the generation of a relatively small proportion of the Ig heavy chain genes expressed by normal adult B cells. The noted restriction in the use of Ig V(H) genes by AIDS-associated B-cell lymphomas suggests that antigenic stimulation contributes to lymphomagenesis in patients with AIDS. PMID- 8704182 TI - Laminin promotes differentiation of NB4 promyelocytic leukemia cells with all trans retinoic acid. AB - The promyelocytic leukemia cell line, NB4, carries the t(15; 17) translocation and undergoes limited maturation in response to differentiation agents. Growth on laminin enhanced the ability of all-trans retinoic acid (ATRA) to promote morphologic maturation of these cells. Although exposure to ATRA in suspension yielded minimal maturation beyond the myelocyte stage, after 72 hours of exposure to ATRA on laminin the cells acquired the histologic appearance of metamyelocytes, band forms, and segmented neutrophils. After 96 hours, some cells acquired a spindle shape and became tightly adherent. Growth on collagen types I, III, IV, or fibronectin did not have this effect, although some cells did adhere to fibronectin. NB4 cells treated with ATRA in suspension or on laminin acquired the equivalent ability to reduce nitroblue tetrazolium or cytochrome C. Despite the improved morphologic maturation on laminin, the cells did not express secondary granule proteins such as lactoferrin or neutrophil collagenase. In addition, growth on laminin abolished cell proliferation in the presence of ionomycin. Growth on laminin and/or with ATRA induced new expression of alpha 6 integrin, a laminin receptor, as assessed by reverse transcription-polymerase chain reaction. Different conditions of growth (laminin or differentiation agent) resulted in specific patterns of expression of the alpha 6A and alpha 6B isoforms. Treatment with ATRA also resulted in the acquisition of high-level surface expression of alpha 6 integrin, as assessed by flow cytometry. Thus, treatment of NB4 promyelocytic leukemia cells with ATRA induced expression of alpha 6 integrin (a laminin receptor alpha-chain) and enabled more advanced maturation when the cells were grown on the extracellular matrix component, laminin, compared with tissue culture plastic. PMID- 8704183 TI - Expression of the retinoblastoma protein in low-grade B-cell lymphoma: relationship to cyclin D1. AB - The product of the retinoblastoma tumor-suppressor gene (pRB), a nuclear phosphoprotein that regulates transcription factors such as E2F, is involved in cell cycle control and differentiation. Its activity is regulated by phosphorylation; the underphosphorylated form inhibits transcription whereas the highly phosphorylated form is inactive. Cyclin D1 and its associated kinase (CDK 4/6) phosphorylate pRB in vitro, and therefore are thought to contribute to the regulation of pRB function. To examine the effect of cyclin D1 overexpression on pRB in primary tumor tissue, we studied pRB expression in low-grade B-cell neoplasms, with particular regard to mantle cell lymphoma, which is characterized by cyclin D1 (bcl-1) overexpression. pRB expression was studied by immunostaining with a well-characterized anti-pRB antibody; the phosphorylation status of pRB was examined by immunoblots; and the functional binding capacity of pRB was examined by in vitro binding to adenovirus E1A protein. We studied 3 reactive lymph nodes, 28 low grade B-cell lymphomas, 4 cases of hairy cell leukemia (HCL) and 3 plasmacytomas. Reactive lymph nodes showed intense pRB staining of germinal centers, with strongest (2+) staining in the large cells (centroblasts) of the proliferating (dark) zone and weak or no staining of small lymphocytes, including those of the mantle zone. In B-chronic lymphocytic leukemia (B-CLL) (4 cases), follicular lymphoma (3 cases) and mucosa-associated (MALT) lymphoma (3 cases) strong (2+) pRB staining was limited to centroblasts in reactive and neoplastic follicles and occasional proliferation centers, with only faint staining of small lymphoid cells. In contrast, 15 of 16 cases of mantle cell lymphoma showed strong (1-2+) staining of most cells; one blastoid mantle cell lymphoma showed only faint pRB staining. All cases of (HCL) and plasmacytoma showed strong pRB staining. Although most lymphomas with strong pRB expression were cyclin D1(+), three cyclin D1(+) cases showed only weak pRB expression (1 B-CLL, 1 blastoid mantle cell, 1 unclassifiable low grade B-cell lymphoma). Conversely, of the 4 pRB(+) HCLs and 3 pRB(+) plasmacytomas, only 1 of each was cyclin D1(+). pRB appeared to exist primarily in the underphosphorylated (fastest migrating) form on Western blot, despite the fact that cyclin D1 was complexed to CDK4, a form in which it normally phosphorylates pRB. In addition, pRB appeared to be unmutated, because it bound normally to the adenovirus E1A protein and showed nuclear localization by immunostaining. We conclude that most cases of mantle cell lymphoma, HCL, and plasmacytoma show high levels of pRB in contrast to follicle center lymphoma and small lymphocytic lymphoma; however, pRB expression does not appear to be consistently related to cyclin D1 overexpression. The pRB appears to be unmutated and underphosphorylated, and therefore should be in its active form. Our data from primary lymphoma tissue suggests that overexpression of cyclin D1, whereas tumorigenic, does not lead to pRB loss or hyperphosporylation. Thus, the mechanism by which cyclin D1 contributes to tumorigenesis and the significance of the restricted expression of pRB in low-grade lymphoid neoplasms remain to be determined. PMID- 8704185 TI - Clonal circulating cells are common in plasma cell proliferative disorders: a comparison of monoclonal gammopathy of undetermined significance, smoldering multiple myeloma, and active myeloma. AB - The blood of most patients with active multiple myeloma (MM) contains cells related to the bone marrow tumor. However, identifying clonal cells in the blood of patients with monoclonal gammopathy of undetermined significance (MGUS) has been difficult. In this study, we analyzed blood mononuclear cells (BMNCs) from 16 patients with MGUS, 2 with amyloidosis, 8 with smoldering MM (SMM), 2 with indolent MM (IMM), and 15 with active MM using three different methods to detect and quantitate clonal cells, ie, immunofluorescence microscopy (IM) for monoclonal plasma cells, three-color flow cytometry (FC) for CD38(+)CD45- CD45(dim) cells, and the allele-specific oligonucleotide polymerase chain reaction (ASO-PCR). Using ASO-PCR, we were able to detect clonal cells in the blood in 13 of 16 patients with MGUS, 2 of 2 with amyloid, 6 of 8 with SMM, 2 of 2 with IMM, and 13 of 15 with MM. In 9 of the 13 patients with MGUS with blood involvement, the number of clonal cells was very small ( < 0.04% of the BMNCs). The median percentage of clonal cells as determined by ASO-PCR was 0.02 for MGUS, 0.02 for SMM, and 0.24 for MM. Clonal plasma cells or CD38+CD45-CD45(dim) cells were identified by IM or FC in 6 of 16 MGUS patients, 4 of 8 with SMM, and 11 of 15 with MM. In all cases in which IM or FC detected clonal cells, the ASO-PCR was positive. This study shows that, by using ASO-PCR, clonal cells can be found at very low levels in the blood in most patients with MGUS. However, the number of clonal cells in the blood of MGUS patients is less than those with overt MM (P = .006). In contrast to MGUS, patients with active MM are more likely to have identifiable clonal circulating plasma cells (P = .05). PMID- 8704184 TI - Identification of novel K562 membrane proteins that adhere to bone marrow fibroblasts. AB - Adhesion of myeloid leukemia cells to the bone marrow (BM) microenvironment is mediated in part by Beta 1 and Beta 2 integrins. Cells of the erythroleukemia line K562, derived from a patient with chronic myeloid leukemia, bind to BM fibroblasts (BMFs) but the adhesion cannot be accounted for by integrins or other known adhesion proteins including CD44 or members of the Ig or selectin families. Membrane fragments from K562 cells were radioiodinated and allowed to adhere to BMF monolayers. Adherent proteins were solubilized together with the fibroblasts, analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and visualized by autoradiography. Four adherent proteins were consistently observed. Two of these, with reduced molecular weights of 52 kD and 35 to 37 kD, were prominent. Addition of soluble thrombospondin and heparin but not fibronectin inhibited binding of K562 membrane proteins to adherent BMFs and immobilized thrombospondin- and heparin-bound K562 proteins. The 52-kD protein has a multimeric structure nonreduced and has characteristics of a glycoprotein. Its adhesion to fibroblasts is divalent cation and temperature sensitive. The 35- to 37-kD protein, whose function is divalent cation but not temperature sensitive, is phosphoinositol-linked and has characteristics identical to an adherent 35- to 37-kD protein identified on murine progenitor cells. Membrane preparations from two cases of acute myeloid leukemia showed an adherent 35- to 37-kD protein and in one case an adherent 52-kD protein without other adherent bands. A K562 subclone with reduced adherence to BMFs showed reduced amounts of adherent 52-kD and 35- to 37-kD proteins. These proteins may be responsible for the adhesion of malignant and normal hematopoietic progenitor cells to the BM microenvironment. PMID- 8704186 TI - Kaposi's sarcoma (KS)-associated herpesvirus-like DNA sequences in peripheral blood mononuclear cells: association with KS and persistence in patients receiving anti-herpesvirus drugs. AB - Herpesvirus-like DNA sequences (KSHV/HHV-8) have recently been described in AIDS associated Kaposi's sarcoma (KS) lesions. Many questions remain regarding the role of this virus in KS and the therapeutic implications of this finding. In the current study, KSHV/HHV-8 DNA was detected in peripheral blood mononuclear cells (PBMCs) from human immunodeficiency virus (HIV)-infected patients with KS (34/98) more often than in HIV-infected individuals without KS (12/64, P = .03). The detection of KSHV/HHV-8 DNA did not correlate with the CD4 lymphocyte count. Five patients demonstrated KSHV/HHV-8 DNA in their PBMCs during administration of intravenous foscarnet and/or ganciclovir. The continued detection of KSHV/HHV-8 DNA in the PBMCs of patients receiving these anti-herpesvirus drugs has potential implications regarding the virus-cell relationship of KSHV/HHV-8, as well as for the value of these drugs in treating or preventing KS, but additional studies are needed. PMID- 8704187 TI - Idiopathic thrombocytopenic purpura: a practice guideline developed by explicit methods for the American Society of Hematology. PMID- 8704188 TI - TEL-AML1 fusion RNA as a new target to detect minimal residual disease in pediatric B-cell precursor acute lymphoblastic leukemia. AB - It has recently been shown that the t(12;21)(p13;q22) translocation fuses two genes, TEL on chromosome 12 and AML1 on chromosome 21. We have evaluated the frequency of this newly described translocation in acute lymphoblastic leukemia (ALL), and the feasibility of minimal residual disease (MRD) monitoring by polymerase chain reaction (PCR) amplification of TEL-AML1 transcripts. Thirty nine adult- and 45 childhood-ALLs consecutively diagnosed in a single center were included in this study. TEL-AML1 fusion transcripts were searched for in the 39 adult- and 45 childhood-ALLs for which material was available. BCR-ABL, E2A-PBX1, and MLL-AF4 transcripts were also studied by PCR in these cases. TEL-AML1 transcripts were found in 8 out of 35 (23%) childhood B-cell precursor ALLs (BCP ALLs). TEL-AML1 transcripts were detected in only 1 of 31 adult BCP-ALLs (P = .04, Fisher's exact test). Nevertheless, in this adult case, TEL-AML1 transcripts were found at a low level in 2 of 3 different samples. BCR-ABL, E2A-PBX1, and MLL AF4 transcripts were found in 12, 3, and 1 cases of 31 adult BCP-ALLs, and in 1, 2, and 1 cases of 35 childhood BCP-ALLs, respectively. TEL-AML1 transcripts were never found associated with any other fusion transcripts. Taken together, the four types of chimeric transcripts were detected in 12 of 35 (34%) childhood BCP ALL cases. No TEL-AML1 transcripts were detected in 11 T-cell ALLs (4 adults and 5 children), nor in 2 B-cell (slg+) ALLs. MRD was evaluated in 21 samples collected in 9 TEL-AML1+ childhood BCP-ALL cases during therapy (median follow-up = 200 days). Of 8 patients evaluated after induction therapy, 4 showed detectable but low levels of MRD. Of 7 patients serially evaluated, only one showed persistence of detectable MRD. This study shows that TEL-AML1 transcripts are frequently detected in pediatric BCP-ALLs and that these transcripts are molecular targets that will simplify the strategy of MRD monitoring in childhood BCP-ALL. PMID- 8704189 TI - Correlation of P-glycoprotein expression and function in childhood acute leukemia: a children's cancer group study. AB - Clinical drug resistance may be attributed to the simultaneous selection and expression of genes modulating the uptake and metabolism of chemotherapeutic agents. P-glycoprotein (P-gp) functions as a membrane-associated drug efflux pump whose increased expression results in resistance to anthracyclines, epipodophyllotoxins, vinca alkaloids, and some alkylating agents. This type of resistance occurs as both de novo and acquired resistance to therapy for leukemia. We have studied P-gp expression and function in childhood acute leukemias by developing a series of doxorubicin- and vincristine-selected CEM, T cell lymphoblastoid cell lines that recapitulate the low levels of expression and resistance seen clinically. These cell lines have been used to develop flow cytometric assays for the semiquantitative measurements of P-gp expression with the MRK16 monoclonal antibody and P-gp function using the enhanced retention of rhodamine 123 in the presence of verapamil, a resistance modulator. Kolmogorov Smirnov statistics, represented by the D measurement, are used to determine the difference in level of P-gp expression by comparing MRK16 staining to an IgG2a isotype control. When D is > 0.09, there is an excellent correlation (R = 0.82) between P-gp expression and function. The evaluation of 107 bone marrow specimens from 84 children with lymphoblastic or myelogenous leukemia showed a statistically significant (P = .004) increase in P-gp function at relapse. P-gp expression at relapse, however, approached but did not reach a significant level (P = .097). Using this methodology, we can identify patients with levels of P-gp expression and function that we can define clinically, as well as children with discordant multidrug resistance phenotypes. This study supports the role of P-gp mediated drug resistance in childhood leukemia and confirms that P-gp expression and function are measurable in their leukemic blasts. These assays provide the means for the in vitro testing of resistance modulators and the monitoring of in vivo response to treatment with these agents. PMID- 8704190 TI - Role of cytokines in leukemic type growth of myelodysplastic CD34+ cells. AB - The clonal growth of progenitor cells from myelodysplastic syndromes (MDS) can be subdivided into four growth patterns: (1) normal, (2) no growth or low plating efficiency, (3) low colony and high cluster number, and (4) normal or high colony number with a large number of clusters. The former two (1 and 2) can be referred to as nonleukemic patterns and latter two (3 and 4) as leukemic. In a search for a role for cytokines in leukemic-type growth of MDS progenitor cells, marrow CD34+ cells were purified up to 94% for 8 normal individuals and 88% for 12 MDS patients, using monoclonal antibodies and immunomagnetic microspheres (MDS CD34+ cells). The purified CD34+ cells were cultured for 14 days with various combinations of cytokines, including recombinant human macrophage colony stimulating factor (rM-CSF), granulocyte-CSF (rG-CSF), granulocyte-macrophage-CSF (rGM-CSF), interleukin-3 (rIL-3), and stem cell factor (SCF; a ligand for c-kit) in serum-free medium. The clonal growth of MDS CD34+ cells supported by a combination of all of the above cytokines was subdivided into the two patterns of leukemic or nonleukemic, and then the role of individual or combined cytokines in proliferation and differentiation of MDS CD34+ cells was analyzed in each group. Evidence we obtained showed that SCF plays a central role in the leukemic-type growth of MDS CD34+ cells and that G-CSF, GM-CSF; and/or IL-3 synergize with SCF to increase undifferentiated blast cell colonies and clusters over that seen in normal CD34+ cells. SCF is present in either normal or MDS plasma at a level of nanograms per milliliter, and this physiologic concentration of SCF can stimulate progenitor cells. This means that progenitor cells are continuously exposed to stimulation by SCF in vivo and that MDS leukemic cells have a growth advantage over normal blast cells. This depends, at least in part, on cytokines such as G CSF, GM-CSF, IL-3, and SCF. PMID- 8704192 TI - Effect of recombinant granulocyte colony-stimulating factor on neutrophil kinetics in normal young and elderly humans. AB - Recombinant granulocyte colony-stimulating factor (G-CSF) was administered to healthy young (n = 32) and elderly (n = 19) volunteers (0 microgram/d, 30 microgram/d, or 300 microgram/d) to determine its effect on neutrophil production, blood kinetics, and tissue migration. Measurements included blood counts (daily), marrow neutrophil pool sizes and neutrophil tissue migration (baseline and day 5), blood kinetics (day 6), and marrow transit time while on drug (days 6 to 14). G-CSF markedly expanded the marrow neutrophil mitotic pool and shortened the transit time of the postmitotic pool (control, mean = 6.4 days; 300 microgram/d, mean = 2.9 d). G-CSF increased neutrophil production without significantly altering blood neutrophil half-life or margination. Compared to control, neutrophil accumulation in skin chambers decreased by about 50% in the 300 microgram/d group in both young and elderly subjects. G-CSF induced neutrophilia by stimulating proliferation of marrow neutrophil precursors and accelerating neutrophil entry into the blood. Decreased neutrophil inflammatory responses measured with the skin chamber technique may be because of the relative immaturity of the circulating cells or to alterations in neutrophil phenotype induced by G-CSF. PMID- 8704191 TI - Are there functional gap junctions or junctional hemichannels in macrophages? AB - The existence of functional gap junctions in migratory cells of the immune system is a controversial issue. In this report, we have focused on one particular cell type, namely the macrophages, because connexin-43, a protein that forms gap junctions, has been described in peritoneal macrophages and a macrophage cell line (J774), by Northern and Western blot analysis. To test whether these cell types expressed functional gap junctions, we assayed dye coupling by intracellular injection of Lucifer Yellow. We observed that nonstimulated macrophages are not coupled among themselves and did not form functional gap junctions with an epithelial cell line, which expresses functional gap junctions formed by connexin-43. Dye coupling was also not detected between macrophages previously activated by lipopolysaccharide or interferon-gamma. We further examined the presence of functional coupling using the more sensitive technique of dual whole cell patch-clamp, and again, did not find electrical coupling between macrophages, consistent with the dye microinjection data. We also examined the possible presence of hemigap junction channels activated by extracellular adenosine triphosphate (ATP) using a dye uptake assay and the whole cell patch-clamp technique. Conditions expected to close gap junction hemichannels (exposure to octanol and low intracellular pH) did not decrease ATP induced Lucifer Yellow uptake, whereas conditions expected to increase hemichannel opening either did not affect ATP permeabilization (dibutyryl adenosine monophosphate) or decreased it (zero extracellular CA+2). Finally, in experiments using resident macrophages derived from conexin-43 knockout mice, we observed ATP induced dye uptake. Our experimental data thus indicate that macrophages in vitro do not form functional gap junctions and that the permeability pathway activated by extracellular ATP is not formed by a hemigap junction channel. PMID- 8704193 TI - A novel silent posttranslational mechanism converts methionine to aspartate in hemoglobin Bristol (beta 67[E11] Val-Met->Asp). AB - The first reported case of congenital Heinz body hemolytic anemia was subsequently shown to be caused by an unstable hemoglobin, Hb Bristol [beta 67(E11) Val-Asp]. This has become one of the classic models of an unstable hemoglobin, the hydrophilic aspartate disrupting the hydrophobic heme pocket. We have restudied this original case, who remains clinically well after nearly 50 years of severe hemolysis with a hemoglobin level of about 7 g/dL and two unrelated Japanese cases. Surprisingly, all three cases show the same DNA changes, predicting a valine to methionine change at beta 67, rather than the expected aspartate. Further analysis with electrospray ionization mass spectrometry and globin chain biosynthesis strongly suggests that this anomaly is because of a novel posttranslational mechanism, with slow conversion of the translated methionine into an aspartate residue. The proximity of the heme and oxygen may be important in facilitating the reaction. These findings show the importance of complete characterization of variant hemoglobins using protein, DNA, and biosynthetic analyses. PMID- 8704194 TI - Extremely high avidity association of Fe(III) with the sickle red cell membrane. AB - Red blood cells (RBCs) from patients with sickle cell anemia and thalassemia carry abnormal accumulations of molecular Fe(III) at the cytosol/membrane interface. The avidity of the red cell membrane for this iron has not been defined. Using open ghost membranes prepared from sickle RBC, we examined the ability of membrane-associated Fe(III) to resist removal by 15 chelators representing a 40-log range of affinities for Fe(III). Efficacy of chelators was compared with literature values for their idealized affinity for iron as represented by the cummulative stability constant (beta n), their effective stability constant reflecting affinity under biologic conditions (Keff), and an indicator of their ability to chelate Fe(III) in the presence of an insoluble phase of iron (Ksol). Deferoxamine, a very high affinity chelator having log beta n = 30.6, was found to be the lowest affinity chelator able to remove RBC membrane Fe(III). Regardless of chelator beta n, only those agents able to preserve log Keff > or = 12 were able to do so, indicating that the membrane's effective avidity for Fe(III) is on the order of 10(12). Additional confirmation that membrane avidity for Fe(III) is extremely high is found in the observation that only chelators having log Ksol > 0 were effective. Potential physiologic iron chelators in cytoplasm of pathologic red cells are unable to prevent or reverse iron accumulation on the membrane because they do not have sufficiently high affinity for iron. These data argue that RBC membrane Fe(III) is truly pathologic. PMID- 8704196 TI - Factors influencing outcome in de novo myelodysplastic syndromes treated by allogeneic bone marrow transplantation: a long-term study of 71 patients Societe Francaise de Greffe de Moelle. AB - We report on 71 consecutive patients with de novo myelodysplastic syndromes referred to physicians belonging to the Societe francaise de greffe de moelle from 1982 through 1991 and transplanted with marrow from HLA-identical siblings. There were 16 cases of refractory anemia, 27 of refractory anemia with excess of blast cells, and 28 of refractory anemia with excess of blast cells in transformation. Seventeen patients had received cytoreductive chemotherapy before the graft. The disease progressed in 17 patients between diagnosis and grafting. Twenty-three patients are alive with a median follow-up of 6 years, whereas 24 died from relapse and 24 from transplant-related complications. Kaplan-Meier estimates of event-free survival, relapse and transplant-related mortality at 7 years were 32%, 48%, and 39%, respectively. The log-rank test and Cox's model revealed better outcome among young patients, patients in an early stage of the French-American-British (FAB) classification or with a low percentage of marrow blasts before transplantation, patients who did not undergo cytoreductive chemotherapy before transplantation, and patients conditioned with total body irradiation and cyclophosphamide. The high rate of relapse in advanced FAB stages has led us to graft patients earlier in the course of the disease, and we are currently conducting a multicenter, randomized study to determine the value of intensive chemotherapy before grafting in patients with an excess of marrow blasts. PMID- 8704197 TI - Megakaryocyte growth and development factor accelerates platelet recovery in peripheral blood progenitor cell transplant recipients. AB - We have investigated the potential of PEGylated recombinant human megakaryocyte growth and development factor (PEG-rHuMGDF), a molecule related to thrombopoietin (mpl ligand or TPO) in minimizing the thrombocytopenia associated with hematopoietic ablation and peripheral blood progenitor cell (PBPC) transplant. Irradiated mice that received PBPC mobilized by PEG-rHuMGDF or granulocyte colony stimulating factor (G-CSF) had a reduced number of thrombocytopenic days with platelets below 100 x 10(6) per mL of blood. Recipients of unmobilized PBPC had a 9 day thrombocytopenic phase which was shortened to 7 days if they were given granulocyte-macrophage colony-stimulating factor (GM-CSF)-mobilized PBPC. This was further reduced to 2 or 3 days of thrombocytopenia in recipients of G-CSF- or PEG-MGDF-mobilized PBPC. Despite our observation that PEG-rHuMGDF is a relatively modest stimulator of the mobilization of myeloid progenitors to the blood, MGDF mobilized PBPC do effect accelerated recovery of platelets after transplantation. However, the most effective use of PEG-rHuMGDF is when it is given during the recovery phase after PBPC transplantation to hematopoietically ablated mice. Posttransplant treatment with PEG-rHuMGDF reduces thrombocytopenia to a single day or less, in recipients of most types of PBPC. Mice that were treated during the first 2 weeks after PBPC transplant with PEG-rHuMGDF had 1 thrombocytopenic day compared to 9 days in carrier-treated recipients of unmobilized PBPC and 2 to 3 days in carrier-treated recipients of the optimally mobilized PBPC from G-CSF or G-CSF/PEG-rHuMGDF treated donors. In groups where PEG-rHuMGDF was included in the mobilization protocol and used to treat recipients as well thrombocytopenia was effectively eliminated. These data show that PEG-rHuMGDF is a highly effective agent in eliminating the thrombocytopenia associated with PBPC transplantation. PMID- 8704195 TI - Thiotepa cyclophosphamide followed by granulocyte colony-stimulating factor mobilized allogeneic peripheral blood cells in adults with advanced leukemia. AB - Thirty-one patients (median age, 44 years) with advanced hematologic malignancies were given thiotepa 15 mg/kg, and cyclophosphamide 120 (n = 14) or 150 (n = 17) mg/kg followed by unfractionated peripheral blood stem cell transplants (PBSCT) from genotypically identical siblings (n = 28) or one antigen mismatched family donor (n = 3). Donors were mobilized with granulocyte colony-stimulating factor 5 to 10 microgram/kg/d for 6 days and underwent two to three leukapheresis on days +5, +6, +7. The median cell yield per donor expressed/kg of recipients body weight was as follows: nucleated cells 13 x 10(8)/kg; CD34+ cells 6 x 10(6)/kg; colony-forming unit-granulocyte macrophage 38 x 10(4)/kg, and CD3+ cells 449 x 10(6)/kg. The diagnoses were chronic myeloid leukemia (n = 4), acute myeloid (n = 9) or lymphoid leukemia (n = 2), acute myelofibrosis (n = 2), multiple myeloma (n = 1), lymphoma (n = 6), chronic lymphocytic leukemia (n = 1) myelodysplasia (n = 6). Twenty-eight patients had advanced disease, 29 patients were first grafts, and 2 were second transplants 3 and 9 years after the first. Neutrophil counts of 0.5 x 10(9)/L and platelet counts of 30 x 10(9)/L platelets were both achieved on day +14 (median). Engraftment could be proven by sex markers or DNA polymorphism in 29 of 31 patients: one had early leukemia relapse and one patient was unevaluable because of early death. Acute graft-versus-host disease (GVHD) was scored as minimal or absent (grade 0 to 1) in 14 patients, moderate (grade II) in 13, and severe (grade III to IV) in four. Causes of death were leukemia (n = 4), acute GVHD (n = 4, with associated cytomegalovirus infections in three), sepsis (n = 1), liver failure (n = 1), multiorgan failure (n = 1), and hemorrhage (n = 1). The actuarial transplant mortality is 29%, the actuarial relapse rate 22%. Nineteen patients survive with a median follow up of 288 days (100-690). The actuarial 2-year survival is 57%. Three patients received PBSCT from family donors mismatched for one class II antigen: all engrafted, one developed grade I aGVHD; one died of leukemia on day +155; two are alive disease free 267 to 290 days postgraft. This study suggests that thiotepa cyclophosphamide followed by unfractionated PBSC allograft may be an alternative form of transplant for adults with advanced leukemia, also in the setting of one antigen mismatched donor. The engraftment is rapid with acceptable GVHD and relatively low transplant-related mortality. PMID- 8704198 TI - High frequency of acute promyelocytic leukemia among Latinos with acute myeloid leukemia. PMID- 8704199 TI - Humoral immune response limits gene therapy in canine MPS I. PMID- 8704200 TI - Pre-B acute lymphoblastic leukemia cells may induce T-cell anergy to alloantigen. AB - Even if neoplastic cells express tumor associated antigens they still may fail to function as antigen presenting cells (APC) if they lack expression of one or more molecules critical for the induction of productive immunity. These cellular defects can be repaired by physiologic activation, transfection, or fusion of tumor cells with professional APC. Although such defects can be repaired, antitumor specific T cells may still fail to respond in vivo if they may have been tolerized. Here, human pre-B cell acute lymphoblastic leukemia (pre-B ALL) was used as a model to determine if primary human tumor cells can function as alloantigen presenting cells (alloAPC) or alternatively whether they induce anergy. In the present report, we show that pre-B cell ALL express alloantigen and adhesion molecules but uniformly lack B7-1 (CD80) and only a subset express B7-2 (CD86). Pre-B ALL cells are inefficient or ineffective alloAPC and those cases that lack expression of B7-1 and B7-2 also induce alloantigen specific T cell unresponsiveness. Under these circumstances, T-cell unresponsiveness could be prevented by physiologic activation of tumor cells via CD40, cross-linking CD28, or signaling through the common gamma chain of the interleukin-2 receptor on T cells. Taken together, these results suggest that pre-B ALL may be incapable of inducing clinically significant T-cell-mediated antileukemia responses. This defect may be not only due to their inability to function as APC, but also due to their potential to induce tolerance. Attempts to induce clinically significant antitumor immune responses may then require not only mechanisms to repair the antigen presenting capacity of the tumor cells, but also reversal of tolerance. PMID- 8704201 TI - Germ cell defects and hematopoietic hypersensitivity to gamma-interferon in mice with a targeted disruption of the Fanconi anemia C gene. AB - Fanconi anemia (FA) is an autosomal recessive chromosome instability syndrome characterized by progressive bone marrow (BM) failure, skeletal defects, and increased susceptibility to malignancy. FA cells are hypersensitive to DNA cross linking agents, oxygen and have cell cycle abnormalities. To develop an animal model of the disease we generated mice homozygous for a targeted deletion of exon 9 of the murine FA complementation group C gene (fac). Mutant mice had normal neonatal viability and gross morphology, but their cells had the expected chromosome breakage and DNA cross-linker sensitivity. Surprisingly, male and female mutant mice had reduced numbers of germ cells and females had markedly impaired fertility. No anemia was detectable in the peripheral blood during the first year of life, but the colony forming capacity of marrow progenitor cells was abnormal in vitro in mutant mice. Progenitor cells from fac knock-out mice were hypersensitive to interferon gamma. This previously unrecognized phenotype may form the basis for BM failure in human FA. PMID- 8704202 TI - Nonrandom X-inactivation patterns in normal females: lyonization ratios vary with age. AB - The utility of X-inactivation based clonality assays for evaluation of human neoplasia is well-documented. However, excessive Lyonization is a potential limitation of these assays, because it mimics clonal derivation of cells. The incidence of excessive Lyonization in healthy females is controversial, with reported incidence varying from 4% to 33%. Several explanations have been offered for the observed variation, including different criteria for excessive Lyonization, diversity of X-linked clonality assays, small population sizes and more recently, tissue specificity of X-inactivation patterns. However, it is also possible that stem cell depletion, clonal hematopoiesis, or selection pressures on blood cells results in an increased incidence of excessive skewing. If any of the latter is true, then the incidence of excessive skewing should increase with age in blood cells. To test this hypothesis, we determined X-inactivation ratios at the human androgen receptor locus of 295 normal females from three age groups: (1) neonates, (2) females 28 to 32 years old and, (3) females aged > or = 60 years. The incidence of skewing (allele ratios > or = 3:1) was 8.6% (14 of 162) in neonates (P = .104 v 28 to 32); 16.4% (11/67) in 28 to 32 y.o. (P = .0064 v > or = 60), and 37.9% (25 of 66) in women > or = 60 y.o. (P < .0001 v neonates). When a more stringent criterion for skewing was used (allele ratios > or = 10:1), the incidence was 1.9% (3 of 162) in neonates (P = .362 v 28 to 32), 4.5% (3 of 67) in 28 to 32 y.o. (P = .0022 v > or = 60), and 22.7% (15 of 66) in > or = 60 y.o. group (P < .0001 v neonates). Results have been confirmed at the phosphoglycerate kinase locus for 48 heterozygous females. The incidence of excessive skewing increases with age. In neonates, the incidence is low and may correspond to true excessive Lyonization. Acquired skewing occurs with aging in normal females and is present in 38% of females over the age of 60. Further studies are needed to determine whether acquired skewing is a consequence of stem cell depletion, true clonal hematopoiesis, growth advantage conferred by parental specific X-chromosomes, or other causes. These data provide an explanation for variation in reported incidence of excessive skewing in normal females. Furthermore, these findings suggest that any study of clonality using X inactivation based assays should incorporate age-matched controls for Lyonization. PMID- 8704203 TI - Transcriptional regulation of interleukin-3 expression in megakaryocytes. AB - Interleukin-3 (IL-3) is a potent stimulator of megakaryocyte proliferation, and autocrine production of IL-3 by megakaryocytic leukemia cell lines and bone marrow-derived megakaryocytes has recently been demonstrated. To characterize the transcriptional regulation of IL-3 in megakaryocytes, we transiently transfected IL-3 promoter CAT constructs that contain variable amounts of 5' flanking sequences into the human CMK and CMK-6 megakaryocytic cell lines and identified two positive acting transcriptional regulatory regions, one located between bp 315 and -284, which contains consensus AP-1 and ets binding sites, and a second located between bp -173 and -61. DNase I footprinting assays using CMK or CMK-6 nuclear extracts demonstrate DNA-protein interactions in the identical region protected by T cell or natural killer cell nuclear extracts (between bp -165 to 128), and electrophoretic mobility shift assays demonstrate the binding of proteins to three distinct portions of this region. To characterize the transcription factors in megakaryocytic cells that could bind to these two regulatory regions, we performed Northern blot analyses, which showed the presence of ets-1, elf-1 (which is thought to be restricted to T cells), NF-IL3A and AML1 mRNAs, as well as c-fos, jun B, and jun D, but not c-jun mRNA. These studies show that the transcriptional regulation of IL-3 expression in megakaryocytic leukemia cell lines is similar, but not identical to normal human T cells. PMID- 8704204 TI - Interaction of stem cell factor and its receptor c-kit mediates lodgment and acute expansion of hematopoietic cells in the murine spleen. AB - The phenotypes of mice that harbor a defect in the genes encoding either stem cell factor (SCF) or its receptor, c-kit, indicate that this ligand/receptor pair is necessary for maintenance of normal hematopoiesis in the adult. Our objective was to determine whether SCF, like erythropoietin, is necessary for acute erythroid expansion during recovery from hemolytic anemia. Monoclonal antibody ACK2, which recognizes the murine c-kit receptor, was used to selectively block the hematopoietic growth-promoting effects of SCF. Mice were treated with phenylhydrazine on day 0 and day 1 to induce hemolytic anemia and also received no antibody, control IgG, or ACK2 on day 0. The mice were killed on day 3 and the hematocrit (Hct), reticulocyte count, and numbers of erythroid and myeloid hematopoietic progenitor cells (colony-forming unit-erythroid [CFU-E], burst forming unit [BFU]-E, and CFU-granulocyte-macrophage [GM]) were quantitated in the femoral marrow and spleen using hematopoietic colony-forming assays. Induction of hemolytic anemia with phenylhydrazine resulted in a drop in the Hct from approximately 50% to 30%, and an approximate 8- to 10-fold increase in the reticulocyte count. The numbers of CFU-E increased modestly in the femur, and approximately 25- to 50-fold in the spleen, in comparison with normal mice. BFU-E and CFU-GM values did not increase in the femur but expanded 6- to 10-fold in the spleen, in comparison with normal mice. This confirms that much of the erythroid expansion in response to hemolytic anemia occurs in the murine spleen. Neutralizing quantities of the ACK2 antibody reduced femoral CFU-E, BFU-E, and CFU-GM content to less than half that found in phenylhydrazine-treated control mice and nearly totally ablated splenic hematopoiesis. These results suggest that c-kit receptor function may be required for optimal response to acute erythropoietic demand and that erythropoiesis in the splenic microenvironment is more dependent on SCF/c-kit receptor interaction than is erythropoiesis in the marrow microenvironment. Because expansion of late erythropoiesis in the spleen was preferentially blocked, we tested the hypothesis that homing of more primitive hematopoietic cells to the spleen was dependent on c-kit receptor function. Lethally irradiated mice were injected with marrow cells obtained from mice that had received phenylhydrazine plus control IgG or with marrow cells obtained from mice that had received phenylhydrazine plus ACK2. In parallel experiments, normal murine marrow cells were treated in vitro with control IgG or with ACK2 and were injected into lethally irradiated mice. The fraction of BFU-E and CFU-GM retrieved from the marrow and spleen of the recipient mice 4 hours later was reduced by approximately 75% when progenitor cells had been exposed to ACK2, in comparison with control IgG. These data suggest that interaction of SCF with the c-kit receptor affects the homing behavior of hematopoietic progenitor cells in the adult animal. PMID- 8704205 TI - Transforming growth factor beta 1 directly and reversibly inhibits the initial cell divisions of long-term repopulating hematopoietic stem cells. AB - Hematopoiesis appears to be regulated, in part, by a balance between extracellular positive and negative growth signals. Transforming growth factor beta-1 (TGF-beta 1) has been shown to be a negative regulator of primitive hematopoietic cells. This study examined the direct effect of TGF-beta 1 on the proliferation and differentiation of long-term repopulating hematopoietic stem cells (LTR-HSC) in vitro. We previously reported a cell fractionation approach that includes the selection of low Hoescht 33342/low Rhodamine 123 (low Ho/Rh) cell fractions that are highly enriched for long-term repopulating cells (LTR HSC) and also clone to a very high efficiency in the presence of stem cell factor (SCF) + interleukin-3 (IL-3) + IL-6: 90% to 100% of individually cultured low Ho/Rh cells formed high proliferative potential clones. This high cloning efficiency of an LTR-HSC enriched cell population enabled proliferation inhibition studies to be more easily interpreted. In this report, we show that the continuous presence of TGF-beta 1 directly inhibits the cell division of essentially all low Ho/Rh cells (in a dose-dependent manner) during their 0 to 5th cell division in vitro. Therefore, it follows that TGF-beta 1 must directly inhibit the proliferation of LTR-HSC contained within these low Ho/Rh cells. The time required for some low Ho/Rh cells to undergo their first cell division in vitro was also prolonged in the presence of TGF-beta 1. Furthermore, when low Ho/Rh cells were exposed to TFG-beta 1 for varying lengths of time before neutralization of the TGF-beta 1 by monoclonal antibody, the ability to form macroclones was markedly decreased after approximately 4 days of TGF-beta 1 exposure. In addition, 1 to 10 ng/mL of TGF-beta 1 resulted in a maintenance of high proliferative potential-colony-forming cell (HPP-CFC) during 8 days of culture compared with loss of HPP-CFC in cultures with no added TGF-beta 1. In conclusion, this study shows that TGF-beta 1 directly inhibits the initial stages of proliferation of LTR-HSC and appears to slow the differentiation of daughter cells of low Ho/Rh cells. PMID- 8704206 TI - Bone marrow collected 14 days after in vivo administration of granulocyte colony stimulating factor and stem cell factor to mice has 10-fold more repopulating ability than untreated bone marrow. AB - We have examined the repopulating ability of bone marrow and peripheral blood cells collected immediately and at intervals after treatment of donor mice with the combination of granulocyte colony-stimulating factor (G-CSF) and stem cell factor (SCF). Using a competitive repopulation assay we showed that the repopulating ability of peripheral blood cells was highest immediately after cytokine treatment and declined to normal levels within 6 weeks of the termination of treatment with G-CSF and SCF. In contrast the repopulating ability of bone marrow cells was low immediately after cytokine treatment and increased to levels that were 10-fold or more greater than marrow from untreated mice by 14 days after termination of treatment with G-CSF and SCF. This high level of repopulating activity declined to normal levels by 6 weeks after termination of treatment with G-CSF and SCF. The high level of repopulating ability was confirmed by injecting cells from G-CSF- and SCF-treated donors into unconditioned recipients. Peripheral blood cells collected immediately after treatment with G-CSF and SCF engrafted into unconditioned mice sevenfold better than an equivalent number of bone marrow cells from untreated mice. Likewise, bone marrow cells collected 14 days after treatment of the donor animal with G CSF and SCF engrafted at 10-fold higher levels than an equivalent number of bone marrow cells from untreated mice. We conclude that the treatment of donor mice with G-CSF and SCF causes a transient increase in the repopulating ability of peripheral blood and later of bone marrow. These observations may have applications to clinical hematopoietic stem cell transplantation. PMID- 8704207 TI - Induction of negative hematopoietic regulators by neurokinin-A in bone marrow stroma. AB - The tachykinins are a family of neuropeptides that share a common carboxyl terminus. Substance P (SP) and neurokinin-A (NK-A) are derived from the preprotachykinin l gene. Although SP and NK-A can bind to either NK-1, NK-2, or NK-3 receptors (R), they have preferences for NK-1R and NK-2R, respectively. We have reported that SP stimulates erythroid (E) (burst-forming unit [BFU]-E and colony-forming unit [CFU]-E) and myeloid (CFU-granulocyte-macrophage [GM]) progenitors partly through the induction of growth factors. We have now investigated the hematopoietic effects of NK-A using short-term bone marrow (BM) cultures and found that NK-A (10(-7) to 10(-12) mol/L) inhibits CFU-GM proliferation but stimulates erythroid progenitors. Release of soluble factors by the stroma appears to mediate the inhibition because direct contact with the stroma was not required. We have found that NK-A, through NK-2-like receptors induces increased levels of macrophage inflammatory protein-1 alpha (MIP-1 alpha) and transforming growth factor-beta (TGF-beta) (transcriptional and posttranscriptional) in BM stroma. Clonogenic assays with NK-A (10(-9) mol/L) and either anti-MIP-1 alpha or anti-TGF- beta 1 indicate that these cytokines partly contribute to the inhibition, suggesting that these two negative hematopoietic regulators exert part of the inhibition by NK-A on CFU-GM. The findings of two closely related neuropeptides, derived from the same gene, exerting opposite effects on myeloid colonies suggest that neuropeptides, by themselves could be important factors in hematopoietic regulation. PMID- 8704208 TI - Antisense oligodeoxynucleotide combination therapy of primary chronic myelogenous leukemia blast crisis in SCID mice. AB - The proliferation of chronic myelogenous leukemia (CML) cells and the transformation of normal hematopoietic cells by BCR-ABL appear to require the expression of a functional MYC protein, suggesting an approach to treatment of Philadelphia leukemias based on simultaneous targeting of BCR-ABL and c-MYC. To test this hypothesis, CML-blast crisis (CML-BC) primary cells were treated in vitro with bcr-abl and c-myc antisense phosphorothioate oligodeoxynucleotides ([S]ODNs), individually or in combination. Compared with antisense ODNs targeting of individual oncogenes, downregulation of both BCR-ABL and c-MYC by specific antisense [S]ODNs resulted in a synergistic antiproliferative effect. Colony formation of normal bone marrow cells was not affected by either treatment. To assess the therapeutic potential of multiple oncogene downregulation, SCID mice injected with CML-BC primary cells were treated systematically with equal doses of bcr-abl or c-myc antisense [S]ODNs or with a combination of both antisense [S]ODNs. Compared with mice treated with individual compounds, the disease process was significantly retarded in the group treated with both [S]ODNs as revealed by flow cytometry, clonogenic assay, and RT-PCR analysis to detect leukemic cells in mouse tissue cell suspensions. These effects correlated with a markedly increased survival of leukemic mice treated with both antisense [S]ODNs. Leukemic cells harvested from antisense [S]ODN-treated mice were sensitive to the effects of antisense [S]ODNs in vitro, suggesting that the treatment can be successfully repeated. These data demonstrate the therapeutic potential of targeting multiple cooperating oncogenes. PMID- 8704209 TI - Prognostic factors in agnogenic myeloid metaplasia: a report on 195 cases with a new scoring system. AB - We studied the survival of 195 patients with agnogenic myeloid metaplasia (AMM) diagnosed between 1962 and 1992 in an attempt to stratify patients into risk groups. Median survival was 42 months. Adverse prognostic factors for survival were age > 60 years, hepatomegaly, weight loss, low hemoglobin level (Hb), low or very high leukocyte count (WBC), high percentage of circulating blasts, male sex, and low platelet count. A new scoring system based on two adverse prognostic factors, namely Hb < 10 g/dL and WBC < 4 or > 30 x 10(9)/L, was able to separate patients in three groups with low (0 factor), intermediate (1 factor), and high (2 factors) risks, associated with a median survival of 93, 26, and 13 months, respectively. An abnormal karyotype (32 cases of 94 tested patients) was associated with a short survival, especially in the low-risk group (median survival of 50 v 112 months in patients with normal karyotype). The prognostic factors for acute conversion were WBC > 30 x 10(9)/L and abnormal karyotype. Thus, hemoglobin level and leukocyte count provide a simple prognostic model for survival in AMM, and the adverse prognostic value of abnormal karyotype may be related to a higher rate of acute conversion. PMID- 8704210 TI - The effect of the Fanconi anemia polypeptide, FAC, upon p53 induction and G2 checkpoint regulation. AB - Fanconi anemia (FA) is an autosomal recessive disease marked by developmental defects, bone marrow failure, and cancer susceptibility. FA cells are hypersensitive to DNA cross-linking and alkylating agents and accumulate in the G2 phase of the cell cycle in response to these agents. FA cells also display genomic instability, suggesting a possible defect in the p53 pathway. To test the effect of heterologous expression of FAC cDNA on drug-induced cytotoxicity, G2 accumulation, and p53 induction in FA cells, we compared two isogenic FA cell lines: HSC536N (mock), a FA type C cell line sensitive to mitomycin C (MMC), and the same cell line transfected (corrected) with wild-type FAC cDNA (HSC536N [+FAC]). HSC536N (+FAC) cells showed a 30-fold increase in resistance to MMC concentration. Similarly, increases in resistance were observed following exposure to cisplatin, carboplatin, and cyclophosphamide. In addition, HSC536N (+FAC) cells showed a twofold lower G2 accumulation following MMC treatment. To analyze the possible interaction of FAC with the p53 pathway, we analyzed p53 induction in mock and corrected cell lines following exposure to MMC. HSC536N (mock) cells induced p53 at lower MMC concentrations than HSC536N (corrected). Caffeine, a known G2 checkpoint inhibitor, not only inhibited G2 accumulation seen in both cell lines but also caused the resistant HSC536N (+FAC) to become as sensitive to MMC as HSC536N (mock) cell line. We conclude that the FAC protein has a specific cytoprotective effect and may function as a cell cycle regulator of the G2 phase of the cell cycle. PMID- 8704211 TI - Frequent clonal loss of heterozygosity but scarcity of microsatellite instability at chromosomal breakpoint cluster regions in adult leukemias. AB - Microsatellites are important highly polymorphic genetic markers dispersed in the human genome. Using a panel of 22 (CA)n repeat microsatellite markers mapped to recurrent breakpoint cluster regions specifically involved in leukemia, we investigated 114 adult leukemias (25 acute lymphocytic leukemia [ALL], 32 acute myeloid leukemia [AML], 36 chronic lymphocytic leukemia [CLL], and 21 chronic myeloid leukemia [CML] in chronic phase) for somatic mutations at these loci. In each patient, DNA from fresh leukemia samples was analyzed alongside normal constitutive DNA from buccal epithelium. We detected loss of heterozygosity (LOH) in 81 of 114 patients (ALL 16/25, AML 25/32, CLL 30/36, CML 10/21). Deletions were most often seen in ALL at 11q23 and 19p13; in AML at 8q22 and 11q23; in CLL at 13q14.3, 11q13, and 11q23; and in CML at 3q26. Only six deletions were reported in 74 karyotypes analyzed, whereas in these same cases, 91 LOH events were detected by microsatellites. Of 26 leukemias with a normal karyotype, 16 nevertheless showed at least one LOH by microsatellite analysis. Replication errors were found in 10 of 114 patients (8.8%). Thus, microsatellite instability is rare in leukemia in contrast to many solid tumors. Our findings suggest that in adult leukemia, LOH may be an important genetic event in addition to typical chromosomal translocations. LOH may point to the existence of tumor suppressor genes involved in leukemogenesis to a degree that has hitherto been underestimated. PMID- 8704212 TI - Interleukin-10 gene expression in adult T-cell leukemia. AB - We studied the serum levels of interleukin-10 (IL-10), in patients with adult T cell leukemia (ATL) caused by human T-cell leukemia virus type I (HTLV-I) infection. Elevated IL-10 levels were observed in 33 of 45 patients with ATL. Fresh leukemic cells from ATL patients as well as HTLV-I-infected T-cell lines MT 2, SLB-1, and C10/MJ expressed IL-10 mRNA by reverse transcription-polymerase chain reaction analysis, whereas IL-10 mRNA was not detected in normal peripheral mononuclear cells and an uninfected T-cell line Jurkat. IL-10 protein was also detected in the culture medium of leukemic cells from ATL patients as well as these HTLV-I-infected cell lines, and in the extracellular fluids of ATL patients. Interestingly, MT-4 cells, which did not express Tax although transformed by HTLV-I, did not express IL-10 at either the mRNA or protein level. To elucidate the role of the HTLV-I encoded transactivator Tax in IL-10 gene expression, Jurkat cells were transfected with a Tax expression plasmid. In transiently transfected Jurkat cells, endogenous IL-10 mRNA expression was induced by Tax. Stably transfected Jurkat cell lines expressed IL-10 mRNA and secreted IL-10 protein into the culture medium. The nuclear factor (NF)-kappa B pathway is a target for Tax transactivation. We treated MT-2 cells with phosphorothioate antisense oligonucleotides to the p65 subunit of NF-kappa B. A reduction in the expression of p65 was accompanied by a reduction in IL-10 gene expression and IL-10 production. We showed that the IL-10 kappa B-like sites ( kappa B1,-2,034 to -2,025; kappa B2, -1,961 to -1,952; kappa B3, -452 to -443) specifically formed a complex with NF-kappa B-containing nuclear extract from MT 2 cells and that NF-kappa B bound with the highest affinity to the kappa B2 element (kappa B2 > kappa B3 > kappa B1). These data suggest a general role for NF-kappa B activation in the induction of IL-10 gene transcription. Activation of IL-10 in HTLV-I-infected cells may contribute to the pathology associated with HTLV-I infection. PMID- 8704213 TI - Prognostic significance of BCL-2 expression and bcl-2 major breakpoint region rearrangement in diffuse large cell non-Hodgkin's lymphoma: a British National Lymphoma Investigation Study. AB - The Bcl-2 protein is capable of preventing apoptosis, and in vitro evidence suggests a role in drug resistance. It is expressed and the gene is rearranged in a proportion of cases of large-cell non-Hodgkin's lymphoma (NHL), but the clinical significance of these findings is controversial. The purpose of this study was to determine the influence of both Bcl-2 expression and major breakpoint region (MBR) bcl-2 rearrangement in a large cohort of prospectively accrued patients with intermediate-grade B-cell NHL treated in a standardized manner. All patients with Working Formulation F, G, or H NHL treated with cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP) chemotherapy in British National Lymphoma investigation studies between July 1974 and April 1992 were considered for this study if the appropriate paraffin blocks were available. Paraffin sections from the diagnostic specimen were analyzed for evidence of MBR rearrangement using a polymerase chain reaction-based method, and for Bcl-2 expression using immunohistochemistry. Failure to achieve complete remission (CR), relapse, death from NHL, and deaths from all causes were used as end points to measure CR rate, actuarial relapse rate, actuarial survival from NHL, and actuarial overall survival. One hundred sixty-one suitable patients were identified and tested for the bcl-2 MBR translocation, with 27 (17%) found to be positive; 153 of these patients were tested with immunocytochemistry, and 84 (55%) showed evidence of Bcl-2 expression. For patients who achieved CR from the initial treatment, the relapse rate was significantly higher in those with Bcl-2 expression than in those without. In addition, multivariate analysis identified Bcl-2 expression as the only factor significantly related to relapse rate in the subjects measured. The cause-specific survival for NHL in the series as a whole was significantly lower in patients with Bcl-2 expression than in those without. MBR status had no significant influence on any of the outcome measures, but the number of MBR-positive patients was relatively small, and larger studies are required. In conclusion, in Working Formulation F, G, and H NHL of B-cell type, expression of Bcl-2 protein predicted independently for relapse. PMID- 8704214 TI - In vitro studies on cellular and molecular mechanisms of arsenic trioxide (As2O3) in the treatment of acute promyelocytic leukemia: As2O3 induces NB4 cell apoptosis with downregulation of Bcl-2 expression and modulation of PML-RAR alpha/PML proteins. AB - It has been shown recently in China that arsenic trioxide (As2O3) is a very effective treatment for acute promyelocytic leukemia (APL). APL patients resistant to all-trans retinoic acid (ATRA) and conventional chemotherapy can still respond to AS2O3. In this study, we addressed the possible cellular and molecular mechanisms of this treatment by using NB4 cells as a model. The results show that: (1) As2O3 triggers relatively specific NB4 cell apoptosis at micromolar concentration, as proved by morphology, histogramic related nuclear DNA contents, and DNA gel eletrophoresis. (2) As2O3 does not influence bax, bcl x, c-myc, and p53 gene expression, but downregulates bcl-2 gene expression at both mRNA and protein levels. (3) As2O3 induces a significant modulation of the PML staining pattern in NB4 cells and HL-60 cells. The micropunctates characteristic of PML-RAR alpha in NB4 cells dissappear after treatment with As2O3, whereas a diffuse PML staining occurs in the perinuclear cytoplasmic region. In addition, a low percentage of untreated NB4 cells exhibits an accumulation of PML positive particles in a compartment of cytoplasm. The percentage of these cells can be significantly increased after As2O3 treatment. A similar PML staining pattern is observed in apoptotic cells. (4) ATRA pretreatment does not influence As2O3-induced apoptosis. These results suggest that induction of cell apoptosis can be one of the mechanisms of the therapeutic effect of As2O3. Moreover, this apoptosis induction occurs independently of the retinoid pathway and may be mediated, at least partly, through the modulation of bcl-2, as well as PML-RAR alpha and/ or PML proteins. PMID- 8704215 TI - Hereditary spherocytosis with band 3 deficiency. Association with a nonsense mutation of the band 3 gene (allele Lyon), and aggravation by a low-expression allele occurring in trans (allele Genas). AB - We describe an 18-year-old with moderate hereditary spherocytosis. The condition was associated with a 35% decrease in band 3. The underlying mutation was Arg to stop at codon 150 (CGA-->TGA) and was designated R150X, which defined allele Lyon of the EPB3 gene. The inheritance pattern was dominant. However, the mother, who also carried the allele Lyon, had a milder clinical presentation and only a 16% decrease of band 3. We suggested that the father had transmitted a modifying mutation that remained silent in the heterozygous state. Nucleotide sequencing after single strand conformation polymorphism analysis of the band 3 cDNA and promoter region revealed a G-->A substitution at position 89 from the cap site in the 5'-untranslated region, designated 89G-->A, which defined allele Genas. A ribonuclease protection assay showed that (1) the allele Genas (father) resulted in a 33% decrease in the amount of band 3 mRNA, (2) the reduction caused by the allele Lyon (mother) was 42%, and (3) the compound heterozygous state for both alleles (proband) resulted in a 58% decrease. These results suggest that some mildly deleterious alleles of the EPB3 gene are compensated for by the normal allele in the heterozygous state. They are shown through the aggravation of the clinical picture, based on more obvious molecular alterations when they occur in trans to an allele causing a manifest reduction of band 3 membrane protein concentration. PMID- 8704216 TI - Inhibition of hemoglobin expression by heterologous production of nitric oxide synthase in the K562 erythroleukemic cell line. AB - Recent studies have indicated that nitric oxide may affect iron metabolism through disruption of the iron-sulfur complex of iron regulatory protein-1, a translational regulator. In the present study, we report that heterologous expression of murine macrophage nitric oxide synthase (NOS-2) in the human erythroleukemic K562 cell line results in constitutive production of nitric oxide associated with inhibition of hemoglobin expression. K562 cells were transfected with an episomally-maintained, hygromycin-selectable expression vector bearing the coding region of NOS-2. Constitutive NOS expression was detected by Western blotting of cell lysates and by the accumulation of nitrite in the culture media. Although NOS-transfected cells grew more slowly than control cells, they were able to maintain constitutive expression of NOS and production of nitric oxide for more than 1 month following transfection. The hemoglobin content of NOS transfected K562 cells was less than one-fifth that of control cells, but increased markedly if NOS inhibitor was included in the culture media. The nitric oxide-mediated inhibition of hemoglobin expression was reversed by supplementing the culture media with 20 mumol/L hemin or 0.5 mmol/L 5-amino-levulinate, indicating that nitric oxide did not directly inhibit hemoglobin synthesis, but likely acted on a step in heme synthesis. mRNA levels for globin and erythroid aminolevulinic acid synthase (eALAS) were the same in both NOS-transfected and control cells. Our observations indicate that hemoglobin expression is inhibited by nitric oxide in NOS-transfected K562 cells by posttranscriptional repression of eALAS, the first enzyme of the heme biosynthetic pathway. The most likely mechanism is a nitric oxide-mediated translational repression of eALAS, as was recently demonstrated for ferritin synthesis. These observations further illustrate the potential for endogenously produced nitric oxide to regulate cellular posttranscriptional events. In particular, our observations may be relevant to the role of nitric oxide in anemia and lowered blood hemoglobin concentrations that are associated with chronic infections, such as tuberculosis or parasitic disease. PMID- 8704218 TI - Active involvement of catalase during hemolytic crises of favism. AB - The endemic occurrence of favism in certain Mediterranean regions provided an investigative opportunity for testing in vivo the validity of claims as to the role of catalase in protecting human erythrocytes against peroxidative injury. Reduced activity of catalase was found in the erythrocytes of six boys who were deficient in erythrocytic glucose-6-phosphate dehydrogenase (G6PD) and who were studied while suffering hemolysis after ingesting fava beans. Activity of catalase was further reduced when their red blood cells were incubated with aminotriazole. In contrast, minimal reduction of catalase activity was found, both with and without incubation with aminotriazole, in erythrocytes of a G6PD deficient boy who had ingested fava beans 7 days earlier and in erythrocytes of seven G6PD-deficient men with a past history of favism. These results confirmed earlier studies in vitro indicating that catalase is a major disposer of hydrogen peroxide in human erythrocytes and, like the glutathione peroxidase/reductase pathway, is dependent on the availability of reduced nicotinamide adenine dinucleotide phosphate (NADPH). The effect of divicine on purified catalase and on the catalase of intact G6PD-deficient erythrocytes was similar to the previously demonstrated effect on catalase of a known system for generating hydrogen peroxide. This effect of divicine strengthens earlier arguments that divicine is the toxic peroxidative component of fava beans. PMID- 8704217 TI - In vivo search for butyrate responsive sequences using transgenic mice carrying A gamma gene promoter mutants. AB - We describe an in vivo approach, in transgenic mice, aimed to identify promoter elements responsible for the induction of gamma globin expression by butyrate. Transgenic lines carrying human A gamma gene promoter truncations at position 141, -201, -382, and -730 A gamma were treated with alpha amino butyric acid (alpha ABA), and effects on gamma globin expression were analyzed at the messenger RNA level. No induction of gamma gene expression was observed in animals carrying promoters truncated at positions -141, -201, or -382 A gamma, suggesting either that butyrate response elements (BRE) are not located in the proximal gamma gene promoter or, if they were, they require the cooperation of upstream sequences for gamma gene induction. Two animals from one line carrying the -730 A gamma truncation responded to alpha ABA treatment with significant increases in gamma gene expression, indicating that a BRE is located between position -382 and -730 region of the A gamma gene promoter. Because the maximum induction by alpha ABA is observed in transgenic mice carrying a A gamma gene promoter extending to nucleotide -1350, it is likely that another butyrate responsive element is located between -730 and -1350 of the A gamma gene promoter. These results indicate that the transgenic mouse model can be used for identification of DNA regions that contain cis elements involved in gamma globin gene inducibility. PMID- 8704219 TI - Reconstruction of the immune system after unrelated or partially matched T-cell depleted bone marrow transplantation in children: immunophenotypic analysis and factors affecting the speed of recovery. AB - We prospectively studied immune reconstitution in 102 children who underwent T lymphocyte depleted bone marrow transplants using either closely matched unrelated donors or partially matched familial donors by assaying total lymphocyte counts (TLC), T-cell subsets, B cells, and natural killer cells. TLC, CD3+, and CD4+ T-cell counts remained depressed until 2 to 3 years posttransplant, whereas CD8+ T-cell counts normalized by 18 months, resulting in an inverted CD4:CD8 ratio until 12 months posttransplant. Although the percentage of NK cells was elevated early posttransplant, their absolute numbers remained normal. CD20+ B cells were depressed until 12 to 18 months posttransplant. Factors affecting immunophenotypic recovery were analyzed by nonparametric statistics. Younger patients tended to have higher TLC posttransplant. Higher marrow cell doses were not associated with hastened immunophenotypic recovery. Graft-versus-host disease (GVHD) and/or its treatment significantly delayed the immune reconstitution of CD3+, CD4+, and CD20+ cells. The presence of cytomegalovirus was associated with increased CD8+ counts and a decrease in the percentages of CD4+ and CD20+ cells. PMID- 8704220 TI - Human HLA-specific T-cell clones with stable expression of a suicide gene: a possible tool to drive and control a graft-versus-host- graft-versus-leukemia reaction? AB - Allogeneic bone marrow transplantation is still limited by the morbidity and mortality caused by graft-versus-host disease (GVHD), resulting from host recognition by donor T lymphocytes. It is possible to drastically reduce the T cell content of the graft. However, transplanted T cells can also have a beneficial effect by graft enhancement and the graft-versus-leukemia effect. How can we keep the beneficial GVL effect while protecting the patient from possible GVHD? A recent report proposed the ex vivo transfer of the herpes simplex thymidine kinase (HSv-tk) gene into donor T cells before their infusion with hematopoietic stem cells. This procedure is expected to allow selective donor T cell depletion with ganciclovir should GVHD occur, but it has two major drawbacks: reinjection of a fraction of untransfected T cells cannot be avoided and heterogeneity of the transfected population results in increased risks such as HSv-tk gene instability or dysfunction of some of the transfected T cell. Alternative approaches must be considered. We demonstrate here the feasibility of generating HSv-tk transfected HLA-specific CD4+ cytotoxic T-cell clonal populations, in which 100% of the cells have the HSv-tk gene inserted at a single site within their genome. These clones retained their specificity, their function, and their sensitivity to ganciclovir treatment. Our approach is not limited to bone marrow transplantation. Indeed, this procedure represents a useful alternative to retroviral gene transduction and is applicable to every circumstance where clinical use of gene modified T-cell clones is to be considered. PMID- 8704221 TI - Granulocyte colony-stimulating factor recruitment of CD34+ progenitors to peripheral blood: impaired mobilization in chronic granulomatous disease and adenosine deaminase--deficient severe combined immunodeficiency disease patients. AB - Peripheral blood (PB) CD34+ cells mobilized by granulocyte colony-stimulating factor (G-CSF) administration are potentially useful for transplantation and as a target of gene transfer for therapy of hematopoietic disorders. Efficient harvest and planning for clinical use of PB CD34+ cells ideally requires foreknowledge of the expected mobilization kinetics and yield. We developed a sensitive flow cytometric assay for accurately enumerating CD34+ cells throughout the range seen at baseline to peak mobilization. We used this assay to assess the kinetics of G CSF-mediated mobilization of CD34+ cells to PB in normal volunteers and in patients with chronic granulomatous disease (CGD) or adenosine deaminase (ADA) deficient severe combined immunodeficiency disease (SCID). Two dose levels of G CSF were examined (5 and 10 micrograms/kg/d for 7 days). Both doses were well tolerated. For normal subjects and patients an increase in PB CD34+ cells was first detected only preceding the third dose of G-CSF (day 3), peaked transiently on day 5 or 6, and then decreased thereafter despite additional doses of G-CSF. With 32 normal volunteers mean peak CD34+ cell counts were 57 and 76 cells/mm2 of blood (5 and 10 micrograms doses, respectively), whereas for 18 CGD patients the mean peaks were 31 and 40 cells/mm2 of blood. For 2 ADA-deficient SCID patients studied at a G-CSF dose of 5 micrograms/kg/d, the average peak was 16 cells/mm2 of blood. For both of these patient groups mobilization of CD34+ cells to PB was impaired compared with similarly treated normal subjects (P < .05). By contrast to the kinetics of the CD34+ cell mobilization, the absolute neutrophil count (ANC) increased markedly by 6 hours after the first dose of G-CSF and then increased steadily through day 8. At days 5 and 6 (peak mobilization of CD34+ cells) the mean ANC of CGD and ADA patients was only slightly lower ( < or = 15%) than that seen with normal subjects, whereas the difference in CD34+ cell mobilization was > 48%. Thus, ANC is not a reliable surrogate to predict peak PB CD34+ cell counts and direct enumeration of PB CD34+ counts should be undertaken in decisions regarding timing and duration of apheresis to harvest a specific number of these cells. Finally, unexpected, but significant differences in the PB CD34+ cell mobilization between normal subjects and patients with inherited disorders can occur and underscores the importance of establishing the expected mobilization of PB CD34+ cells in the planning of treatment approaches using these cells. PMID- 8704223 TI - Monoclonal gammopathy in patients with chronic hepatitis C virus infection. PMID- 8704222 TI - Interferon-gamma suppresses T-cell proliferation to mitogen via the nitric oxide pathway during experimental acute graft-versus-host disease. AB - The development of graft-versus-host disease (GVHD) is associated with long lasting and profound deficits in immune function that lead to increased morbidity and mortality after bone marrow transplantation (BMT). We investigated a mechanism of T-cell immunodeficiency in response to mitogen or alloantigen in an experimental model of acute GVHD by analyzing the roles of two immunosuppressive moieties: interferon gamma (IFN-gamma) and nitric oxide (NO). Splenocytes from mice with GVHD did not proliferate either to the T-cell mitogen, concanavalin A (Con A), or to host alloantigens, but only mitogen-activated cultures produced increased levels of NO. The abrogation of NO synthesis with LG-mono-methyl arginine (NMMA) restored mitogen-induced proliferation but not the response to host antigens. The mechanism of impared proliferation to mitogen was dependent on IFN-gamma because blockade of this cytokine in culture inhibited NO production and restored proliferation to Con A to levels similar to those in transplanted control mice without GVHD. NMMA did not substantially reduce IFN-gamma levels, demonstrating that NO acted distally to IFN-gamma in the pathway of immunosuppression in response to mitogen. Furthermore, the prevention of IFN gamma production in vivo after allogeneic BMT, by transplantation of polarized type 2 donor T cells (secreting interleukin-4 but not IFN-gamma), also prevented NO production and restored splenocyte responses to mitogen. Our data demonstrate the existence of NO-dependent and NO-independent pathways involved in suppression of T-cell proliferation during acute GVHD. Excess NO synthesis appears to be one mechanism by which IFN-gamma induces immunodeficiency after allogeneic BMT. PMID- 8704224 TI - Fas (CD95) receptor and Fas-ligand expression in bone marrow cells from patients with myelodysplastic syndrome. PMID- 8704225 TI - Growth of Epstein-Barr virus-associated B-lymphoproliferative disease tissue in a severe combined immunodeficient mouse. PMID- 8704226 TI - Circulating clonal lymphocytes in myeloma determined by mRNA in situ hybridization. PMID- 8704228 TI - Retraction. Blockage of the early events of mitogenic signaling by interferon gamma in macrophages in response to colony-stimulating factor-1. PMID- 8704227 TI - Risk of hepatitis C virus infection, Waldenstrom's macroglobulinemia, and monoclonal gammopathies. PMID- 8704229 TI - Molecular analysis of the granulocyte colony-stimulating factor receptor. PMID- 8704230 TI - In vivo adenovirus vector-mediated transfer of the human thrombopoietin cDNA maintains platelet levels during radiation-and chemotherapy-induced bone marrow suppression. AB - Thrombopoietin (TPO, c-mpl ligand) has emerged as a major hematopoietic cytokine stimulating megakaryocyte proliferation, endomitosis, and platelet production. This study shows that a single administration of an adenovirus (Ad) vector encoding TPO (AdCMV.TPO) abrogates thrombocytopenia induced in mice by carboplatin and irradiation. Normal Balb/c mice receiving the vector had increased platelet counts peaking at 7 days and returning to baseline by day 15. Mice rendered pancytopenic with 500 rads and 1.2 mg of carboplatin had a nadir platelet count of five percent of the baseline. Mice receiving AdCMV.TPO 3 days before receiving irradiation and chemotherapy achieved a platelet nadir fourfold higher, and had significant reduction in duration of thrombocytopenia, than mice receiving the control Ad vector. Introduction of AdCMV.TPO the same day of chemotherapy and irradiation was equally effective in acceleration of platelet recovery, but administration of AdCMV.TPO 3 days after chemotherapy-radiation had little effect on platelet recovery. At 30 days after therapy bone marrow and spleen of mice treated with AdCMV.TPO were populated with a large number of polyploid megakaryocytes, but there was no evidence of circulating megakaryocytes in the liver or lungs and no pathologic bone abnormalities such as osteosclerosis or myelofibrosis. These observations suggest that an Ad vector may be an excellent delivery system to provide adequate TPO production to maintain platelet levels in circumstances associated with life-threatening thrombocytopenia. PMID- 8704231 TI - TEL-AML1 translocations with TEL and CDKN2 inactivation in acute lymphoblastic leukemia cell lines. AB - The t(12;21) (p 13; q22) results in the fusion of the TEL gene located on chromosome 12 with the AML1 gene located on the derivative chromosome 21. Because this translocation is difficult to detect using standard cytogenetic techniques, 27 previously karyotyped B-lineage acute lymphoblastic leukemia (ALL) cell lines were evaluated for the presence of the TEL-AML1 fusion using the reverse transcriptase-polymerase chain reaction (RT-PCR), fluorescence in situ hybridization (FISH), and cDNA sequencing. Six cell lines expressed the TEL-AML1 chimeric transcript by RT-PCR and the t(12;21) was confirmed by FISH analysis with probes for TEL, AML1, and chromosome 12. While only one of the 6 cell lines with the t(12;21) lost the der(12)t(12;21)-encoded AML1-TEL fusion transcript, 4 cell lines lacked expression of the nontranslocated allele of TEL and 5 cell lines lacked expression of CDKN2. Moreover, in 2 patients (1 with the TEL-AML1 transcript and 1 without), TEL expression was lost with disease progression; le, TEL was expressed in the initial cell lines (established at diagnosis or first relapse) whereas TEL was not expressed in the cell lines established from these patients in late-stage disease. These data show the coexistence of multiple genetic defects in childhood B-lineage ALL Cell lines with t(12;21) will facilitate the study of TEL-AML1 and AML1-TEL fusion proteins as well as TEL and CDKN2 gene inactivation in leukemia transformation and progression. PMID- 8704232 TI - Successful transplantation of HLA-matched and HLA-mismatched umbilical cord blood from unrelated donors: analysis of engraftment and acute graft-versus-host disease. AB - To reduce the morbidity and mortality associated with unrelated donor bone marrow (BM) transplantation and potentially extend the pool of suitable donors, cryopreserved unrelated donor umbilical cord blood was considered as an alternate source of hematopoietic stem cells for transplantation. Patients with leukemia, BM failure syndrome, or inborn error of metabolism were eligible for a phase I clinical trial designed to estimate the risk of graft failure and severe acute graft-versus-host disease after transplantation of umbilical cord blood from unrelated donors. As of December 21, 1995, unrelated donor umbilical cord blood was used to reconstitute hematopoiesis in eighteen patients aged 0.1 to 21.3 years weighing 3.3 to 78.8 kg with acquired or congenital lympho-hematopoietic disorders or metabolic disease. Patients received either HLA-matched (n = 7) or HLA-1 to 3 antigen disparate (n = 11) grafts collected and evaluated by the New York Blood Center (New York, NY). The probability of engraftment after unrelated donor umbilical cord blood transplantation was 100% with no patient having late graft failure to date. The probability of grade III-IV acute graft-versus-host disease at 100 days was 11%. With a median follow-up of 6 months (range, 1.6 to 17 months); the probability of survival at 6 months is 65% in this high risk patient population. We conclude that cryopreserved umbilical cord blood from HLA matched and mismatched unrelated donors is a sufficient source of transplantable hematopoietic stem cells with high probability of donor derived engraftment and low risk of refractory severe acute graft-versus-host disease. Limitations with regard to recipient size and degree of donor HLA disparity remain to be determined. PMID- 8704233 TI - Acute myeloid leukemia, M2 with t8:21 translocation. PMID- 8704235 TI - Constitutive activation of STAT proteins in primary lymphoid and myeloid leukemia cells and in Epstein-Barr virus (EBV)-related lymphoma cell lines. AB - Although various molecular mechanisms of STAT protein (signal transducers and activators of transcription) activation have been identified, little is known about the functional role of STAT-dependent transcriptional activation. Herein we report the constitutive nuclear localization, phosphorylation, and DNA-binding activity of STAT proteins in leukemia cells and lymphoma cell lines. With the use of oligonucleotide probes derived from the Fc gamma RI promoter, the beta-casein promoter and a STAT-binding element in the promoter of the Bci-2 gene constitutive activation of STAT proteins was detected in untreated acute T- and C/B-leukemia cells (3 of 5 and 12 of 19 patients, respectively). Supershift analyses using Stats 1-6 specific antisera showed the constitutive DNA binding activity of Stat5 in these cells. Confocal microscopy revealed the nuclear localization of Stat5 and Western blot analyses showed tyrosine phosphorylation of Stat5 in nuclear extracts of acute leukemia cells. In contrast, peripheral blood mononuclear cells did not display constitutive STAT-DNA interaction. Further studies were performed on freshly isolated acute myeloid leukemia cells as well as on cell line derived K562, lymphoblastoid cells (LCL), and Burkitt's lymphoma cells (BL). Fluorescence microscopy, gelshift, and supershift experiments showed the nuclear localization and constitutive DNA-binding activity of Stat5 in K562 cells. Stat1 and Stat3 were constitutively activated in freshly isolated AML cells (10 of 14 patients) and in Epstein Barr virus-positive or interleukin-10 expressing permanent LCL and BL cells. Thus, these data indicate a differential pattern of STAT protein activation in lymphoid or myeloid leukemia and in lymphoma cells. PMID- 8704234 TI - Low levels of erythroid and myeloid progenitors in thrombopoietin-and c-mpl deficient mice. AB - Thrombopoietin (TPO), the ligand for the c-mpl receptor, has been shown to be the major regulator of platelet production. Mice deficient in either c-mpl or TPO generated by homologous recombination show a dramatic decrease in platelet counts, but other blood cell counts are normal. Because TPO treatment of myelosuppressed mice not only enhances the recovery of platelets but also accelerates erythroid recovery, we investigated the levels of myeloid and erythroid progenitor cells in TPO-or c-mpl-deficient mice. Our results show that the number of megakaryocyte, granulocyte-macrophage, erythroid, and multilineage progenitors are significantly reduced in the bone marrow, spleen, and peripheral blood of either TPO-or c-mpl-deficient mice. Administration of recombinant murine TPO to TPO-deficient mice and control littermate mice significantly increased the absolute number of myeloid, erythroid, and mixed progenitors in bone marrow and spleen. This increase was especially apparent in TPO-deficient mice where numbers were increased to a level greater than in diluent-treated control mice and approached or equaled that in the TPO-treated control mice. Moreover, TPO administration greatly increased the number of circulating progenitors as well as platelets in both TPO-deficient and control mice. Furthermore, the megakaryocytopoietic activity of other cytokines in the absence of a functional TPO or c-mpl gene was shown both in vitro and in vivo. PMID- 8704236 TI - Monocyte function in a severe combined immunodeficient patient with a donor splice site mutation in the Jak3 gene. AB - Janus kinase-3 (Jak3) is a nonreceptor tyrosine kinase functionally coupled to cytokine receptors which share a "common" gamma chain (gamma c). Mutations in gamma c and Jak3 genes have been identified in X-linked and autosomal severe combined immuno deficiency (SCID), respectively. Jak3 is expressed and activated in myelomonocytic cells. The present study was designed to define the structural alteration responsible for lack of Jak3 in a patient with autosomal SCID and to characterize monocyte function in the absence of this signal transduction element, as well as to establish the whole exon-intron structure. Polymerase chain reaction analysis, performed with primers designed on exon sequences, identified 20 exons spanning approximately 15 kb. These primers, or others designed on the flanking sequences provided in the present report, can be used to amplify the whole gene, allowing the definition of the molecular defects in all cases, including prenatal diagnosis, in which transcript analysis is not possible. On this basis, the deletion transcript found at the homozygous state in patient CM, with both his consanguineous parents being heterozygous for the deletion, was associated with mutation (T to C) of a splice donor site of intron 16 that was also detected in his mother's DNA. Monocytes from Jak3-SCID showed normal cytokine production in response to interleukin-4 (IL-4) (release of IL-1 receptor antagonist) and IL-2 (release of tumor necrosis factor-alpha and IL-8). Lipopolysaccharide-induced cytokine production was also normal and was blocked by IL-4 in Jak3- SCID monocytes. Interferon-gamma induced augmented expression of major histocompatibility class II in Jak3-SCID monocytes. These data indicate that Jak3, expressed and activated in myelomonocytic cells, is dispensable for monocyte differentiation and responsiveness to cytokines that interact with gamma c receptors as well as to other regulatory signals. PMID- 8704237 TI - Evaluation of a CD5-specific immunotoxin for treatment of acute graft-versus-host disease after allogeneic marrow transplantation. AB - Acute graft-versus-host disease (GVHD) is most often treated with high dose glucocorticoids, but less than half of patients have durable overall improvement. Previous phase I and phase II studies suggested that treatment with a CD5 specific immunotoxin (XomaZyme-CD5 Plus) could ameliorate symptoms of GVHD. In a randomized, double-blind trial, we compared XomaZyme-CD5 Plus and glucocorticoids versus placebo and glucocorticoids as initial therapy for 243 patients who developed acute GVHD after allogeneic marrow transplantation. The study drug (XomaZyme. CD5-Plus or an identical appearing placebo) was administered at a dose of 0.1 mg/kg body weight on each of 14 consecutive days. All patients were treated concomitantly with a standard regimen of methylprednisolone. At the time of entry on study, 94% of patients had a rash, 56% had hyperbilirubinemia, 61% had diarrhea, and 84% had nausea and vomiting. At 3, 4, and 5 weeks after starting treatment, symptom severity was less in the CD5 group than in the placebo group. At 4 weeks, 40% of patients assigned to the CD5 group had complete response compared with 25% of those assigned to the control group (P = .019). At 6 weeks, 44% of patients assigned to the CD5 group had complete response as compared with 38% in the placebo group (P = .36). Clinical extensive chronic GVHD developed in 65% of patients in the CD5 group compared with 72% in the control group (P = .35). Survival at 1 year after treatment was 49% in the CD5 group and 45% in the control group (P = .68). Side effects required close monitoring and appropriate adjustment of treatment. The combined administration of a CD5 specific immunotoxin and glucocorticoids controls GVHD manifestations more effectively than treatment with glucocorticoids alone during the first 5 weeks after starting treatment. Use of this immunotoxin does not result in any long term clinical benefit for patients with acute GVHD. PMID- 8704238 TI - Effectiveness of intensified rotational combination chemotherapy for late hematologic relapse of childhood acute lymphoblastic leukemia. AB - Relapsed acute lymphoblastic leukemia (ALL) usually carries a dire prognosis. We evaluated the effectiveness and long-term complications of intensive rotational combination chemotherapy for late hematologic relapse (median, 16 months after elective cessation of therapy) among 34 children and young adults (ages 4 to 23 years). Concurrent central nervous system (CNS) relapse was present in 3 cases and testicular relapse in 4. Secondary therapy comprised an intensive five-drug reinduction (6 weeks) followed by continuation treatment with four drug pairs, rotated weekly in 4-week cycles over 120 weeks. Intrathecal chemotherapy (methotrexate, hydrocortisone, cytarabine) was given three times during reinduction and every 8 weeks during continuation. Treatment was electively discontinued at week 120 in the absence of detectable disease. Thirty-three patients (97%) attained a second complete remission. At a median follow-up of 9.3 years (range, 4.5 to 11.4), estimates of 5-year second event-free and overall survival (+/- SE) are 65% +/- 8% and 79% +/- 7%, respectively. Eleven patients had a second relapse (9 marrow, 2 testicular) and one developed secondary myeloid leukemia. There have been no CNS relapses or deaths in remission. Treatment was well-tolerated and was given largely on an outpatient basis. Late effects are primarily endocrinologic; one child had a second malignant solid tumor (presumed related to initial radiation therapy) that was treated successfully. Intensive treatment with alternating non-cross-resistant drug pairs for late hematologic relapses of ALL is effective and well-tolerated, and produces results similar to those achieved in patients with newly diagnosed ALL. Event-free survival compares favorably with reports of other relapse regimens, including those incorporating bone marrow transplantation. PMID- 8704239 TI - Autotransplants in multiple myeloma: what have we learned? AB - Of 496 consecutive patients with multiple myeloma (MM) enrolled in clinical trials of tandem transplants with peripheral blood stem cells support, 470 (95%) completed the first autotransplant with melphalan 200 mg/m2 (MEL 200) and 363 (73%) completed the second transplant with either MEL 200 (40%), MEL 140 mg/m2 (MEL 140) with total-body irradiation (17%), or a combination of alkylating agents (16%), depending on the response status prior to the second transplant; 31 patients up to age 60 years received an allograft as the second transplant. The median interval from first to second transplant was 5 months. Treatment-related mortality during the first year after transplantation was 7%, and complete remission (CR) was obtained in 36%; the median durations of event-free survival (EFS) and overall survival (OS) after transplant were 26 and 41 months, respectively. Low beta 2-microglobulin ([B2M] < or = 2.5 mg/L) and C-reactive protein ([CRP] < or = 0.4 mg/dL) were the most significant standard parameters associated with both prolonged EFS and OS. Median OS exceeded 5.5 years in the one third of patients with both low B2M and CRP. When cytogenetics were included in the analysis, the presence of 11q abnormalities and/or complete or partial deletion of chromosome 13 ("unfavorable karyotype") became a dominant negative feature for both EFS and OS. In addition to these pretransplant parameters, attainment of CR and application of two transplants within 6 months both significantly extended EFS and OS. The group of patients (7%) with high B2M and CRP with either IgA isotype or unfavorable karyotype had the worst prognosis (EFS, < or = 10 months; median OS, < or = 12 months) and will require novel therapy. We conclude that tandem transplants are feasible in the majority of patients up to age 70 years, effecting CR in one third of all patients. Median OS was greater than 5.5 years, regardless of pretransplant features, if the first transplant was applied within 12 months of initial treatment and the second transplant no more than 6 months later. PMID- 8704240 TI - Mechanism of transcriptional activation of the immediate early gene Egr-1 in response to PIXY321. AB - Studies with the granulocyte-macrophage colony-stimulating factor (GM CSF)/interleukin-3 (IL-3) fusion protein, PIXY321, demonstrated enhanced biological activity of this molecule in comparison with GM-CSF or IL-3 alone or in combination. Experiments were performed to study the mechanisms resulting in PIXY321-induced egr-1 expression in human myeloid leukemic cells (TF-1). Transfections of egr-1 promoter constructs revealed that PIXY321 stimulation resulted in fourfold induction of the -116 and -600 nucleotide (nt) constructs. We transfected a -116 nt construct containing a deletion of the cyclic AMP response element (CRE) or mutation in the serum response element (SRE) and demonstrated that both the SRE and CRE are necessary for maximal induction. However, PIXY321 stimulation resulted in 2.5-fold induction of a SRE-CRE containing construct (P < .05), suggesting that the SRE and CRE are sufficient for PIXY321 responsiveness. Electrophoretic mobility shift assays (EMSA) revealed that the CRE binding protein (CREB) was phosphorylated on serine 133 in PIXY321 stimulated but not -unstimulated extracts from cells cultured in GM-CSF. By Western analysis and EMSA, CREB was constitutively phosphorylated in TF-1 cells grown on PIXY321 before growth factor and serum starvation. However, in TF-1 cells grown on GM-CSF before starvation, CREB phosphorylation was observed 10 minutes after PIXY321 stimulation. Further-more, ENSAs with PIXY321-stimulated and -unstimulated extracts demonstrated the presence of specific proteins that recognize the SRE. Our data demonstrate that transcriptional regulation of egr-1 by PIXY321 is mediated by the CRE and SRE. PMID- 8704241 TI - Fibronectin improves transduction of reconstituting hematopoietic stem cells by retroviral vectors: evidence of direct viral binding to chymotryptic carboxy terminal fragments. AB - Efficient transduction of reconstituting hematopoletic stem cells (HSC) is currently only possible by cocultivation of target cells directly on producer cell lines, a method not applicable to human gene therapy protocols. Our laboratory has previously shown adhesion of primitive hematopoletic stem and progenitor cells to the carboxy-terminal 30/35-kD fragment of the extracellular matrix molecule fibronectin (FN 30/35) (Nature 352:438, 1991) and increased transduction of human hematopoietic progenitor cells via retroviral vectors while adherent to this fragment (J Clin Invest 93:1451, 1994). Here we report that (1) transduction of reconstituting murine HSC assayed 12 months after infection with retrovirus supernatant on FN 30/35 is as effective as cocultivation directly on producer cells; (2) recombinant retrovirus particles directly adhere to FN 30/35 in a quantitative and dose-dependent fashion; and (3) increased transduction efficiency on FN 30/ 35 does not appear to be associated with increased cell proliferation or activation of protein phosphorylation typically induced by integrin-fibronectin interactions. Therefore, we speculate that supernatant infection of HSC on FN 30/35 leads to colocalization of retrovirus particles and target cells on FN 30/35 molecule with a large increase in local virus titer presented to the cell. These findings have direct and important implications for the modification of current human gene therapy protocols. PMID- 8704242 TI - Direct synergistic effects of leukemia inhibitory factor on hematopoietic progenitor cell growth: comparison with other hematopoietins that use the gp130 receptor subunit. AB - Because leukemia inhibitory factor (LIF) has little or no effect on murine hematopoietic progenitor cell growth yet enhances hematopoiesis in vivo, we sought to determine whether the effects of LIF were directly or indirectly mediated, or a combination of both. Although LIF alone or in combination with granulocyte-macrophage colony-stimulating factor (GM-CSF) or interleukin-3 (IL-3) has no effect on colony formation of unfractionated bone marrow cells (BMCs), it enhances M-CSF-induced colony formation. In comparison, LIF synergizes with IL-3, GM-CSF, M-CSF, and Steel Factor (SLF) to promote the colony formation of partially purified lineage-negative (Lin-) BM progenitors without altering their differentiation. These effects were directly mediated since identical results were observed in single-cell assays. Comparing the effect of LIF with other members of this subclass of hematopoietins (IL-6, oncostatin M [OSM], and ciliary neurotrophic factor [CNTF]), we found that while LIF and IL-6 equally synergize with M-CSF and SLF to promote the colony formation of Lin- BMCs, OSM, and CNTF have no effect. In agreement with OSMs ability to directly bind gp130, preincubation of BMCs with OSM inhibits progenitor cell growth stimulated by the combination of LIF or IL-6 plus SLF. LIF can also directly enhance the growth of further purified more primitive Lin- c-kit+ progenitor cells in the presence of IL-3, GM-CSF, or SLF. Thus, LIF can directly synergize with growth factors to promote the proliferation of purified hematopoietic progenitors, suggesting that the direct effects of LIF on hematopoietic cell growth can, in part, explain the observed hematopoietic effects in vivo. This is a US government work. There are no restrictions on its use. PMID- 8704243 TI - Restoration of thrombolytic potential in plasminogen-deficient mice by bolus administration of plasminogen. AB - Homozygous plasminogen-deficient (Plg-/-) mice had a significantly reduced thrombolytic capacity toward intravenously injected 125I-fibrin labeled plasma clots prepared from Plg-/- murine plasma (9% +/- 3% lysis after 8 hours; (mean +/ SEM, n = 6), as compared with 82% +/- 8% in wild-type mice; P < .0001). Bolus injection of 1 mg purified murine plasminogen in 10- to 17-week-old Plg-/- mice increased the plasminogen antigen and activity levels at 8 hours to normal levels (130 +/- 5 micrograms/mL). Plasminogen administration was associated with significant restoration of thrombolytic potential (64% +/- 7% spontaneous clot lysis; P < .0001 versus lysis without plasminogen injection). Bolus injection of 1 mg plasminogen in homozygous tissue-type plasminogen activator-deficient (t-PA /-) mice doubled the plasminogen antigen and activity levels after 8 hours and increased 125I-fibrin clot lysis at 8 hours from 13% +/- 3% to 34% +/- 5% (P = .008). Fibrinogen, t-PA antigen and alpha 2-antiplasmin activity levels after 8 hours were not significantly different in the groups with or without plasminogen injection. Injection of plasminogen induced a variable increase (on average 7- to 10-fold) of PAI-1, but no correlation with the extent of spontaneous clot lysis was observed. Histopathologic examination at the end of the experiments revealed that fibrin deposition in the liver of Plg-/- mice was slightly reduced 8 hours after bolus plasminogen injection (P = .007) and markedly reduced after 24 hours (P < .0001). Plasminogen antigen levels in liver extracts were comparable with those found in wild-type mice at 8 hours (130 +/- 20 versus 110 +/- 15 ng/mg protein) and decreased to 25 +/- 3.2 ng/mg protein at 24 hours. Thus, restoration of normal plasminogen levels in Plg-/- mice normalized the thrombolytic potential toward experimentally induced pulmonary emboli, and resulted in removal of endogenous fibrin deposits within 24 hours. PMID- 8704244 TI - Venous thromboembolism associated with double heterozygosity for R506Q mutation of factor V and for T298M mutation of protein C in a large family of a previously described homozygous protein C-deficient newborn with massive thrombosis. AB - It is remarkable that certain patients with heterozygous protein C (PC) deficiency manifest venous thromboembolism (VTE), whereas others, particularly those belonging to families with homozygous PC deficiency, remain asymptomatic. The goals of the present study of a family, in which the proband had homozygous PC deficiency, were to identify members with and without VTE, to determine the mutation causing PC deficiency, and to search for the R506Q mutation of factor V (FV) causing activated PC resistance. Heterozygosity for a T298M mutation in exon 9 of the PC gene was found in the father of the homozygous proband who died of massive thrombosis. Based on analysis of a three-dimensional molecular model of PC, we speculate that this mutation causes type I deficiency due to disruption of packing of hydrophobic side chains and loss of an H-bond between Q184 and T298. Forty-six family members were examined for the T298M mutation by polymerase chain reaction (PCR) amplification of exon 9 and restriction analysis using Mae III and for the FV R506Q mutation by PCR amplification of exon 10 and restriction analysis using Mnl I. VTE was observed in five of 11 members who were heterozygous for both PC and FV mutations. In contrast, VTE was not observed for the PC mutation in 13 heterozygotes who had normal FV, including the parents of the deceased proband, 10 heterozygotes for the FV mutation who had normal PC, and 12 individuals bearing neither mutation. These observations extend recent evidence of an increased thrombotic risk conferred by the coexistence of heterozygous PC deficiency and heterozygous activated PC resistance and support the paradigm in which hereditary thrombophilia is often a multigenic disease. PMID- 8704245 TI - Factor VIIa and antithrombin III activity during severe sepsis and septic shock in neutropenic patients. AB - Septic shock and multiple organ failure may be associated with coagulation activation, disseminated fibrin formation, and consumption of coagulation inhibitors such as antithrombin III. We have evaluated prospectively coagulation measurements in patients with severe chemotherapy-induced neutropenia. This group of patients was chosen because of their high risk of developing severe septic complications, thus allowing serial prospective coagulation testing before and during evolving sepsis or septic shock. Sixty-two patients with febrile infectious events were accrued to the study. Of these, 13 patients progressed to severe sepsis and 13 additional patients to septic shock as defined according to standard diagnostic criteria. At the onset of fever, factor (F) VIIa activity, FVII antigen and antithrombin III (AT III) activity decreased from normal baseline levels and were significantly lower in the group of patients who progressed to septic shock compared with those that developed severe sepsis (medians: 0.3 v 1.4 ng/mL, 21 v 86 U/dL and 45% v 95%; P < .001). The decrease of these measurements in septic shock was accompanied by an increase in prothrombin fragment 1+2 (median: 3.6 v 1.4 nmol/L; P = .05), a marker of thrombin generation. These differences were sustained throughout the septic episode (P < .0001). FVIIa and AT III levels of < 0.8 ng/mL and < 70%, respectively, at onset of fever predicted a lethal outcome with a sensitivity of 100% and 85%, and a specificity of 75% and 85%, respectively. In contrast, FXIIa-alpha antigen levels were not different between groups at onset of fever but increased modestly during the course of septic shock (P = .001). Thus, septic shock in neutropenic patients is associated with increased thrombin generation. Furthermore, both FVIIa and AT III measurements are sensitive markers of an unfavorable prognosis. PMID- 8704246 TI - Distribution of ligand-occupied alpha IIb beta 3 in resting and activated human platelets determined by expression of a novel class of ligand-induced binding site recognized by monoclonal antibody AP6. AB - The sequence beta (3)203-228 is involved, in a yet undetermined manner, in alpha IIb beta 3 function. We now show that murine monoclonal antibody (MoAb) AP6, specific for beta (3)211-221, binds to alpha IIb beta 3 on adenosine diphosphate (ADP)-activated platelets only when the receptor is occupied by intact fibrinogen. The ligand-induced binding-site reported by AP6 is unique in that it is not expressed following occupancy by either RGD peptides or the gamma-chain carboxy-terminal dodecapeptide. Binding of AP6 to platelets coincides temporally with the binding of the MoAb 9F9, specific for a receptor-induced binding site on fibrinogen. Thus, AP6 reports the binding of fibrinogen to the recognition pocket of alpha IIb beta 3. Its binding to thrombin-stimulated washed platelets correlates with secretion as determined using an MoAb to P-selectin. When ultrathin sections of nonactivated platelets were examined by immunogold staining and electron microscopy, AP6 identified a pool of alpha IIb beta 3 colocalizing with P-selectin and suggesting the presence of alpha IIb beta 3-ligand complexes in the alpha-granule membrane. There was little binding of AP6 to surface alpha IIb beta 3 of unstimulated platelets. After ADP-induced activation, AP6 was abundantly distributed over the entire platelet surface, including pseudopods, but only when fibrinogen was present in the medium. ADP had little effect on AP6 reactivity within platelets. This contrasted with washed platelets and thrombin, where extensive AP6 binding was observed within internal membrane pools as early as 10 to 15 seconds after stimulation. Surface labeling with AP6 followed slower kinetics. Flow cytometry on Triton X-100 permeabilized fixed platelets confirmed AP6 binding to alpha IIb beta 3 within the platelet. Thus, our results provide evidence of (1) a pool of alpha-granule alpha IIb beta 3 occupied by ligand in nonactivated platelets; (2) thrombin-induced activation of alpha IIb beta 3 within the platelet, and (3) thrombin-induced mobilization of ligand-bound alpha IIb beta 3 to the surface. PMID- 8704247 TI - Evolution of blood coagulation activators and inhibitors in the healthy human fetus. AB - Blood coagulation proteins were determined in 285 healthy fetuses from 19 to 38 weeks' gestation and compared with those of 60 normal full-term newborns and 40 adult controls. Prolongation of the coagulation screening tests, prothrombin time, activated partial prothrombin time, and thrombin clotting time, in fetuses throughout intrauterine life was explained by low levels of vitamin K-dependent factors (II, VII, IX, and X), contact factors (XI, XII, prekallikrein, and high molecular-weight kininogen), factor V, factor VIII, and fibrinogen. Low levels of antithrombin III, heparin cofactor II, protein C and protein S, and tissue factor pathway inhibitor were also found, and these probably contributed to a satisfactory hemostatic balance. Some of these parameters were evaluated by both immunologic and functional assays to detect possible "fetal" proteins. An increase in factor levels was observed after the thirty-fourth week of intrauterine life for most of the coagulation activators and inhibitors, but only factors V and VIII reached adult values at birth. This study therefore showed that fetal hemostasis is a dynamic system that evolves gradually toward the neonatal state and then toward the adult state. PMID- 8704248 TI - Analysis of GPIIb/IIIa receptor number by quantification of 7E3 binding to human platelets. AB - A large number of glycoprotein (GP) IIb/IIIa receptors are present on the surface of platelets. Studies to define precisely the number of GPIIb/IIIa receptors using specific monoclonal antibodies (MoAbs) or fibrinogen binding have, however, yielded varying estimates of receptor number. To refine the quantitative estimation of GPIIb/IIIa receptors on resting platelets, we have used the MoAb 7E3, which has high affinity for GPIIb/IIIa. Quantitative binding studies were performed using radiolabeled conjugates of 7E3 IgG, as well as fragments and derivatives of 7E3. For platelets obtained from any single individual, the numbers of 7E3 F(ab')2 and IgG molecules bound per platelet were equivalent (approximately 40,000), whereas the number of Fab molecules bound per platelet was consistently approximately twofold higher (approximately 80,000). To investigate the basis of the quantitative disparity in binding of intact 7E3 and 7E3 F(ab')2 versus 7E3 Fab, we studied the binding of a newly constructed, bispecific (Fab')2 molecule containing only a single 7E3 combining site. Because this construct bound to the same extent as the Fab species, the larger size of the intact 7E3 and 7E3 F(ab')2 molecules could not explain the reduced number of molecules that bound per platelet compared to the Fab fragment. Rather, it appears that the valency of the antibody is the critical factor determining the number of antibody molecules bound per platelet. Thus, we conclude that the binding of 7E3 Fab corresponds most closely with surface GPIIb/IIIa number and that the number of GPIIb/IIIa receptors is approximately 80,000 per platelet. PMID- 8704249 TI - Molecular characterization of a glycosylphosphatidylinositol-linked ADP ribosyltransferase from lymphocytes. AB - Mono ADP-ribosyltransferases catalyze the transfer of the ADP-ribose moiety of nicotinamide adenine dinucleotide (NAD) to proteins. It was reported by Wang et al (J Immunol 153:4048, 1994) that incubation of mouse cytotoxic T lymphocytes (CTL) with NAD resulted in the ADP-ribosylation of membrane proteins and inhibition of cell proliferation and cytotoxicity. Treatment of CTL with phosphatidylinositol-specific phospholipase C (PI-PLC) before incubation with NAD prevented the inhibitory effects of NAD on the cells, consistent with the removal of a glycosylphosphatidylinositol (GPI)-anchored ADP-ribosyltransferase on the lymphocyte surface. We have identified and cloned a GPI-linked ADP ribosyltransferase from Yac-1 mouse T-cell lymphoma cells. The deduced amino acid sequence of the Yac-1 transferase was 70% and 41% identical to those of the rabbit skeletal muscle and chicken heterophil, respectively. It contained three noncontiguous sequences similar to those found in several of the bacterial toxin and vertebrate ADP-ribosyltransferases. Based on crystallography of the bacterial toxins, these regions are believed to form, in part, the catalytic site consistent with a common mechanism for the ADP-ribose transfer reaction. In rat mammary adenocarcinoma (NMU) cells transformed with the Yac-1 transferase cDNA, transferase activity was present on the cell surface and was released into the medium by treatment of cells with PI-PLC. Thus, we have cloned a novel gene that has properties identical to the transferase detected in CTL, and may be involved in the NAD-dependent regulation of proliferation and cytotoxicity. PMID- 8704250 TI - Tumor necrosis factor-alpha induces activation of coagulation and fibrinolysis in baboons through an exclusive effect on the p55 receptor. AB - Tumor necrosis factor-alpha (TNF-alpha) can bind to two distinct transmembrane receptors, the p55 and p75 TNF receptors. We compared the capability of two mutant TNF proteins with exclusive affinity for the p55 or p75 TNF receptor with that of wild type TNF, to activate the hemostatic mechanism in baboons. Both activation of the coagulation system, monitored by the plasma levels of thrombin antithrombin III complexes, and activation of the fibrinolytic system (plasma levels of tissue-type plasminogen activator, and plasminogen activator inhibitor type I), were of similar magnitude after intravenous injection of wild type TNF or the TNF mutant with affinity only for the p55 receptor. Likewise, wild type TNF and the TNF p55 specific mutant were equally potent in inducing neutrophil degranulation (plasma levels of elastase-alpha 1-antitrypsin complexes). Wild type TNF tended to be a more potent inducer of secretory phospholipase A2 release than the p55 specific TNF mutant. Administration of the TNF mutant binding only to the p75 receptor did not induce any of these responses. We conclude that TNF Induced stimulation of coagulation, fibrinolysis, neutrophil degranulation, and release of secretory phospholipase A2 are predominantly mediated by the p55 TNF receptor. PMID- 8704251 TI - Cystamine inhibits human immunodeficiency virus-1 replication in cord blood derived mononuclear phagocytes and lymphocytes. AB - The effects of cystamine on the human immunodeficiency virus (HIV-1) expression in cord blood monocytes-derived macrophages (CBMDM) and lymphocytes were investigated. Cystamine suppressed HIV-1 expression in CBMDM and lymphocytes in a concentration-dependent fashion as determined by HIV-1 reverse transcriptase (RT) activity. This inhibitory effect of cystamine occurred with all five HIV-1 strains (both laboratory adopted and fresh isolates) tested in the study. The addition of cystamine to cultures of HIV-1 chronically infected CBMDM also suppressed 80% to 90% of RT activity in comparison with untreated controls. Cystamine also decreased HIV-1 protein expression in CBMDM as determined by indirect immunofluorescence assay. The inhibitory effects of cystamine on HIV-1 did not appear to be caused by toxicity to CBMDM or lymphocytes because there was no change in cell viability or cellular DNA synthesis as evaluated by trypan blue dye exclusion and [3H]-thymidine incorporation at doses of cystamine that inhibit the virus. HIV-1 infected CBMDM or lymphocyte cultures (without cystamine treatment) demonstrated giant syncytium formation or cytopathic effect (CPE), respectively, whereas cystamine-treated cultures lacked the giant syncytia or CPE induced by HIV-1 infection. Thus, these observations indicate that cystamine may have the potential to limit HIV-1 replication in monocytes/macrophages and lymphocytes in vivo and may represent a potentially useful compound in the treatment of pediatric HIV-1 infection and acquired immunodeficiency syndrome. PMID- 8704252 TI - Recirculation and homing of lymphocyte subsets: dual homing specificity of beta 7 integrin(high)-lymphocytes in nonobese diabetic mice. AB - The beta 7-integrin subunit can pair with two alpha-chains, alpha 4 and alpha E, and is expressed mainly on lymphocytes. As an alpha 4-heterodimer it binds to the mucosal addressin MAdCAM-1, thus acting as a mucosal homing receptor. As an alpha E-heterodimer it binds to E-cadherin and is mainly found on intestinal intraepithelial lymphocytes. Consequently, beta 7 is mostly expressed on lymphocytes of the mucosal immune system. To study the compartmentalization of these cells further we compared the distribution of such lymphocytes in two strains of mice (BALB/c and NOD) and found that the distribution of beta 7 positive lymphocytes among various lymphoid tissues in these strains was very different. In NOD mice a conspicuous population of beta 7-integrin(high) lymphocytes expressing either alpha 4, alpha E, or both, was found in nonmucosal lymphoid tissues such as peripheral lymph nodes (PLNs). They mostly expressed the PLN homing receptor L-selectin, and included both naive and memory cells on the basis of their expression of CD44/pgp-1 and CD45RB, as did the few beta 7(high) lymphocytes in BALB/c PLNs. Their homing to Peyer's patches (PPs) and PLNs was equally effective and the cells homing to PPs and PLNs were equal in their level of L-selectin and alpha 4/beta 7 expression. However, functional studies indicated that their homing to PPs mostly depended on alpha 4/beta 7-integrin, whereas they mainly used L-selectin to home to PLNs. beta 7(high) lymphocytes were found also in circulating blood of unmanipulated NOD mice, and their L selectin expression was higher than in BALB/c mice. These results show that lymphocytes of the mucosal immune system may also express the peripheral node homing receptor L-selectin during their recirculation and that in NOD mice they frequently retain a dual homing specificity, which leads to their accumulation in nonmucosal tissues. PMID- 8704253 TI - Decreased interleukin-12 (IL-12) from activated cord versus adult peripheral blood mononuclear cells and upregulation of interferon-gamma, natural killer, and lymphokine-activated killer activity by IL-12 in cord blood mononuclear cells. AB - Interleukin-12 (IL-12) is a critical cytokine regulating natural killer (NK) and T-cell function. We hypothesized that the impaired ability of cord blood (CB) to produce normal adult levels of IL-12 in response to stimulation may contribute to the immaturity of CB immunity. Furthermore, exogenous IL-12 may compensate for the immaturity in CB cellular immunity and have the potential for immunotherapy post cord blood transplantation. We compared the expression and production of IL 12 from activated cord versus adult mononuclear cells (MNC), regulatory mechanisms associated with IL-12 expression in CB MNC, and the effects of IL-12 on induction of CB interferon (IFN)-gamma production, NK, and lymphokine activated killer (LAK) cytotoxicity. Northern analysis and enzyme-linked immunosorbent assay were performed in lipopolysaccharide (LPS)-stimulated CB and adult peripheral blood (APB) MNC. IL-12 mRNA expression was induced within 6 hours with LPS (10 micrograms/ml) and reached peak levels at 12 hours in both CB and APB MNC. However, IL-12 mRNA expression and protein accumulation in CB MNC were 35.8% +/- 4.84% (12 hours, n = 11, P < .05), and 17.6% +/- 1.7% (24, 72, 96 hours, n = 9, P < .05) respectively, when compared with APB MNC. Nuclear run-on assays showed no differences between CB and APB MNC in both the basal levels of transcription and the degree of transcriptional activation. However, the half life of IL-12 p40 mRNA was approximately threefold lower in activated CB MNC than in activated APB MNC (CB: 114 +/- 3.0 minutes v APB: 353 +/- 7.8 minutes, n = 3, P < .05). Exogenous IL-12 (10 U/mL) induced a significant increase of IFN-gamma from both CB and APB MNC (24 hours, 72 hours, P < .05, n = 3). The stimulated CB IFN-gamma level reached comparable levels produced by unstimulated APB. IL-12 treatment also significantly enhanced CB NK cytotoxicity against K562 and NB-100 cell lines to the comparable levels of APB (P < .05, n = 4). CB MNC was more responsive to IL-12 stimulation with respect to IFN-gamma production, NK, and LAK cytotoxicity when compared with APB. The present study suggests that IL-12 mRNA and protein expression is decreased in activated CB. This discrepancy in IL-12 production is secondary, at least in part, to the altered posttranscriptional regulation. The impaired, ability of CB MNC to produce IL-12 in response to stimulation may contribute to the decrease in IFN-gamma production and NK cytotoxicity. However, IL-12 enhanced IFN-gamma and NK activity in CB MNC up to the comparable levels of APB MNC. These findings suggest that reduced expression and production of IL-12 from activated CB may contribute to the immaturity in CB cellular immunity and contribute, in part, to decreased graft-versus-host disease following CB stem cell transplantation. PMID- 8704254 TI - Tumor vaccination with macrophage colony-stimulating factor-producing Lewis lung carcinoma in mice. AB - Expression of various cytokines by cytokine gene-transduced tumor cells has been shown to increase antitumor immunity of tumor-bearing hosts. In the present study, macrophage-colony stimulating factor (M-CSF) cDNA was retrovirally transfected into Lewis lung carcinoma cells (3LL) of C57BL/6 mouse origin, and the effects of M-CSF expression were studied by inoculating syngeneic C57BL/6 mice with M-CSF-expressing 3LL cells. The mice inoculated with the lowest M-CSF producing 3LL clone showed significant prolongation of the survival compared with wild-type 3LL-Inoculated mice, and 70% or more of the mice inoculated with 3LL clones with higher M-CSF production rejected inoculation. Mice injected with radiation-inactivated M-CSF-expressing 3LL cells before or after Inoculation of wild-type 3LL cells showed prolonged survival compared with mice injected with radiated control 3LL cells before or after transplantation of wild-type cells. In vivo depletion of effector subpopulations by injection of antibodies against CD4+ T cells, CD8+ T cells, or natural killer (NK) cells suggested involvement of NK cells and CD4+ T cells in M-CSF-mediated antitumor cytotoxicity in M-CSF producing 3LL cells-inoculated mice. Severe combined immunodeficiency (SCID) mice with defective T- and B-cell function showed prolonged survival duration after inoculation with M-CSF-expressing 3LL cells compared with those transplanted with control 3LL cells, and this effect of M-CSF expression by 3LL-cells in SCID mice was also abolished by in vivo depletion of NK cells by antibody injection. These findings together with the previous reports that M-CSF augments antibody dependent and-independent antitumor cytotoxicity suggest that M-CSF induces tumor immunity in this cytokine-expressing tumor-transplantation model. PMID- 8704255 TI - Prevention of allogeneic marrow graft rejection by donor T cells that do not recognize recipient alloantigens: potential role of a veto mechanism. AB - Clinical trials and experimental studies have demonstrated that donor T cells can play a critical role in preventing allogeneic marrow graft rejection. Results of a previous study showed that donor T cells were most effective for preventing rejection when they recognize an alloantigen expressed by recipient T cells and can cause graft-versus-host disease (GVHD). The present study examined models where marrow graft rejection can be prevented by donor T cells that do not recognize host alloantigens and cannot cause GVHD. Donor T cells prevented rejection of major histocompatibility complex (MHC) class I and II-disparate F1 marrow in parental recipients prepared with > or = 800 cGy total body irradiation (TBI) but not in those prepared with < or = 750 cGy TBI. In recipients prepared with high TBI exposures, rejection was mediated entirely by host CD8 cells. With lower TBI exposures, rejection was mediated by host CD4 cells and CD8 cells. These observations suggested the hypothesis that donor T cells prevent rejection mediated by host effectors that recognize donor MHC class I alloantigens but do not prevent rejection mediated by host effectors that recognize donor class II alloantigens. Consistent with this hypothesis, further experiments showed that F1 donor T cells can prevent rejection of MHC class I-disparate marrow in irradiated parental recipients but have no detectable effect on rejection of MHC class II disparate marrow. We propose that the expression of MHC class I molecules on donor T cells makes it possible for these cells to inactivate the host response against donor class I alloantigens through a veto mechanism, whereas the absence of MHC class II molecules on murine T cells explains why these cells cannot inactivate the host response against donor class II alloantigens. Finally, donor CD4 cells and CD8 cells were equivalently effective for preventing rejection of F1 marrow in parental recipients, suggesting that veto activity is not restricted solely to the CD8 subset of murine T cells. A veto mechanism could enable donor T cells to prevent allogeneic marrow graft rejection without causing GVHD. PMID- 8704256 TI - Modulated expression of notch1 during thymocyte development. AB - The Notch gene family encodes transmembrane proteins that have been implicated in control of diverse cellular differentiation events in the fly, frog, and mouse. Mammalian Notch1 is expressed at high levels in thymus and is mutated in a subset of human T-cell acute lymphoblastic neoplasms, suggesting a role in T-cell differentiation. To investigate the patterns of expression of NOTCH1 protein in thymocytes of the developing and mature thymus, antibodies raised against NOTCH1 were used to perform immunohistochemical and flow cytometric analyses. Strong staining for NOTCH1 within the fetal murine thymus was observed as early as 13.5 days postcoitum. By 17.5 days postcoitum, preferential staining of superficial cortical thymocytes was observed, with weak staining of developing medulla. Flow cytometric analysis and immunohistochemical staining of flow-sorted cells confirmed that the highest levels of NOTCH1 expression in adult murine thymus were present in immature cortical thymocytes (CD24high, CD4-CD8-). In contrast, NOTCH1 expression was low or absent in more mature cortical thymocytes (CD24low, CD4+CD8+), whereas intermediate levels of expression were observed in CD4+CD8- and CD4-CD8+ cells. These data indicate a dynamic pattern of NOTCH1 expression during T-cell differentiation and suggest that downregulation of NOTCH1 may be required for maturation of cortical thymocytes. PMID- 8704257 TI - Detection and quantification of melphalan-DNA adducts at the single cell level in hematopoietic tumor cells. AB - Bifunctional alkylating agents, such as melphalan, are widely used in the treatment of hematological malignancies. The effects of these drugs on particular types of hematological cells and the causes of treatment failure are poorly understood. The aim of this work was to establish an ability to measure the extent to which melphalan reacts with the DNA of individual tumor cells, thereby creating new possibilities for molecular pharmacological studies on clinical samples. A novel approach for staining drug-DNA adducts is described in which cells were embedded in agarose and then lysed. The DNA from each cell remained in an ideal state for quantitative immunofluorescent staining using a previously described monoclonal antibody. Immunofluorescence and DNA-Hoechst dye fluorescence were quantified using a cooled slow scan charge coupled device camera and image analysis procedures. Immunofluorescence of drug-treated cells from a human leukemia cell line was partially correlated with DNA content. Mean integrated immunofluorescence of 50 to 100 cells was dependent on drug concentration and was linearly related to adduct levels. In these cells and in chronic lymphocytic leukemia cells obtained from patients, there was considerable intercell heterogeneity in apparent adduct levels. This was also seen in peripheral blood mononuclear cells isolated from a patient after melphalan therapy. PMID- 8704258 TI - Application of long-distance polymerase chain reaction to detection of junctional sequences created by chromosomal translocation in mature B-cell neoplasms. AB - Junctional sequences created by chromosomal translocations in mature B-cell neoplasms, which involve immunoglobulin gene loci (IG) and putative proto oncogenes on reciprocal partner chromosomes, are unique to neoplastic cells characterized by particular histological and immunological phenotypes. To establish a rapid and sensitive method to detect neoplastic cells carrying a specific chromosomal translocation, we have developed a novel strategy based on long-distance polymerase chain reaction (LD-PCR) amplification. Genomic DNA was extracted from tumor cells carrying t(14;19)(q32;q13), t(8;14)(q24;q32), t(3;22)(q27;q11), t(2;3)(p12;q27), or t(3;14)(q27;q32). Thirty-two to 35-mer oligonucleotide primer pairs were designed to be complementary to exons or flanking sequences of the BCL3, c-MYC and BCL6 oncogenes, and to IG constant region genes. LD-PCR with a newly available Taq polymerase for longer product synthesis successfully amplified fragments representing BCL3/C alpha junctional sequences for t(14;19); c-MYC/C mu, c-MYC/C gamma, and c-MYC/C alpha for t(8;14); BCL6/C lambda for t(3;22); BCL6/C kappa for t(2;3); 5'-BCL6/C mu, and 5'-BCL6/C gamma for t(3;14). In Burkitt's lymphoma/leukemia, all materials in which c-MYC rearrangements were detectable by conventional Southern blot hybridization showed positive LD-PCR amplification. The sizes of the amplified fragments varied from 1.8 kb to 12 kb, and these were specific to each material. Serial dilution of tumor cells or DNA in negative materials demonstrated a single band on agarose gel electrophoresis stained with ethidium bromide at a level of sensitivity of 10(-3), and hybridization with radioactive probe improved the level by one order of magnitude (1 cell in 10(4)), indicating that this LD-PCR approach is a sensitive technique capable of detecting minimal residual disease. Thus, the present study provided a useful tool for diagnosis and subsequent management of B cell neoplasms characterized by specific chromosomal translocations. PMID- 8704260 TI - Microbiology and computer technology. PMID- 8704259 TI - Neoplastic transformation of normal hematopoietic cells by constitutively activating mutations of c-kit receptor tyrosine kinase. AB - The c-kit proto-oncogene encodes a receptor tyrosine kinase that is crucial to hematopoiesis, melanogenesis, and gametogeneis. Although the enzymatic activity of the c-kit product (KIT) is regulated by its ligand, both the Val559-->Gly (G559) mutation in the juxtamembrane domain and the Asp814-->Val (V814) mutation in the phosphotransferase domain lead to constitutive activation of KIT. By retroviral infection of hematopoietic progenitor cells with KIT(G559) or KIT(V814), KIT(G559) induced development of granulocyte/macrophage and mast-cell colonies in vitro without the addition of exogenous growth factors. KIT(V814) induced factor-independent growth of various types of hematopoietic progenitor cells, resulting in the development of mixed erythroid/myeloid colonies in addition to granulocyte/macrophage and mast-cell colonies. Furthermore, transplantation of KIT(G559) and KIT(V814)-infected bone marrow cells led to development of acute leukemia in one of 10 and six of 10 transplanted mice, respectively. No mice developed hematologic malignancies after transplantation of wild-type KIT-infected cells. Furthermore, transgenic mice expressing KIT(V814) developed acute leukemia or malignant lymphoma. These results demonstrate a direct role of the mutant KITs, particularly KIT(V814), in tumorigenesis of hematopoietic cells and suggest that similar mutations may contribute to the development of human hematologic malignancies. PMID- 8704261 TI - Infection control in the country of the future: report on the first Asian-Pacific International Conference on Hospital Infection--October 25 to 27, 1994. PMID- 8704262 TI - President's message. PMID- 8704263 TI - Designing certification test questions--how you can get involved. PMID- 8704264 TI - Post-discharge surveillance of surgical wound infections. PMID- 8704265 TI - The role of the infection control practitioner. The Community and Hospital Infection Control Association of Canada. PMID- 8704267 TI - [Eye bank in Japan, past-present-future]. PMID- 8704266 TI - A program for reducing biomedical waste: the Wellesley Hospital experience. AB - As hospitals face severe financial constraints, the need to review programs to ensure cost-effectiveness is critical. Waste management practices have come under scrutiny because inappropriate waste disposal is very costly. The Wellesley Hospital in Toronto, Ontario conducted a waste audit in July 1993 and found that 18.5% of waste was being disposed of as biomedical waste. Because the hospital pays nine times more to dispose of biomedical waste than it does landfill waste, a goal was set to reduce biomedical waste to 8% of total waste. A program, which included redefining biomedical waste, reviewing waste practices throughout the hospital, educating staff and monitoring outcomes, resulted in biomedical waste levels decreasing to 7.9% of total waste within 18 months. Savings realized were approximately $67,000. This program is easily reproducible. PMID- 8704268 TI - [Factors related to ADL of stroke patients three months after discharge]. AB - Focusing on the ADL of stroke patients, 94 hemiplegia cases in rehabilitation units of acute care general hospitals were studied to identify factors affecting ADL, measured using the Barthel Index, 3 months after discharge. The main results were as follows: 1. The 8 independent variables: age, gender, length of hospitalization, degree of paralysis of lower extremities at discharge, motivation toward rehabilitation, sight, Barthel scores at discharge, and the expression of one's intentions emerged as the 8 reliable predictive factors (R = .904, R2 = .817). 2. It may be deduced that in older patients in particular, a status of amyotrophy has a significant influence on ADL after discharge. The 94 stroke patients were divided into two age groups-44 patients aged 64 years or younger, and 50 patients aged 65 years or older. Barthel scores at 3 months after discharge were analyzed by multiple regression analysis in both groups. The result was that ADL, after discharge, of the aged 65 years or older group, was more affected by their status at discharge, than was it with the 64 years or younger group (64 years or younger; R = .871, R2 = .758, 65 years or older; R = .934, R2 = .873). 3. For patients in the 64 years or younger group, Barthel scores at 3 months after discharge were significantly related to age, length of hospitalization, sight, Barthel scores at discharge and expression of one's intentions. For patients in the 65 years or older group, the results were significantly related to sight, Barthel scores at discharge, and motivation for rehabilitation. PMID- 8704269 TI - [A survey on infection control practices, knowledge and attitudes toward AIDS/HIV among dental practitioners]. AB - A survey was conducted in December 1993 by sending questionnaires to all 566 dentists of the Iwate Dental Association. The questionnaires consisted of 68 items including infection control practices, knowledge, and attitudes towards AIDS/HIV. The response rate was 51.1 percent (N = 289). The average age of dentists in the sample was 43.7 +/- 9.5 (S.D.) (range: 28 to 85). Data was evaluated statistically by Kruskal-Wallis test, Mann-Whitney's U test and Chi square test, and significant differences were observed. Gloves, masks, and other protective garments were generally worn, but most dentists did not always use them during the full course of treatment limiting usage to surgical treatment, and when treating patients in "high-risk groups". Other infection control procedures, such as instrument sterilization, did not comply with the guidelines of the Ministry of Health and Welfare. Two dentists responded that they had treated AIDS patients and three dentists treated possible AIDS/HIV patients in their dental offices. Despite 71.3% expressing a belief that they have a moral responsibility as a dentist to treat AIDS/HIV patients only 15.6% were willing to treat AIDS/HIV patients. Over 40% of the respondents were not certain as to whether they had treated AIDS/HIV patients or not, and over 60% believed AIDS/HIV patients would come to their dental offices in the near future. Therefore, to practice dentistry on AIDS/HIV patients safely, dentists must provide effective infection control in their dental offices on the assumption that all patients are AIDS/HIV positive, and additional information about AIDS/HIV and adequate training on procedures in the care of AIDS/HIV patients are needed. PMID- 8704270 TI - [Global life satisfaction and its related factors in community elderly residents]. AB - This study was aimed at identifying the distribution of global life satisfaction as measured by Visual Analogue Scale (VAS) and factors related to it in 531 community elderly residents aged 75-80 years. Subjects were divided into two groups according to the extent of their daily activities, which has been emphasized as an important health index for the elderly: the outdoor-activity group (defined as people whose daily activities extended into their community) and the indoor-activity group (defined as people whose daily activities was limited to inside their home). The distribution of global life satisfaction and related factors were compared between the two groups. For independent variables, daily behaviors were classified into five categories and examined for how they related to global life satisfaction. The results obtained were as follows: Global life satisfaction showed a bimodal distribution in both groups tending to be lower in the indoor-activity group than in the outdoor-activity group. Most of the indoor-activity group indicated low or moderate life satisfaction. The extent of their daily activities was thus related to global life satisfaction. The strongest direct factor relating to global life satisfaction was self-rated health for the outdoor-activity group. Some of their daily behaviors also related to it and these significant daily behaviors differed by sex. Only self-rated health was related to global life satisfaction in the indoor-activity group. The results indicate that, when compared to other factors, daily activity in community elderly residents is crucial to increasing their global life satisfaction, and therefore efforts to enlarge subjects' daily activities are also required. PMID- 8704271 TI - [Environmental factors influencing sensitization and onset of Japanese cedar pollinosis among junior high school students]. AB - Japanese cedar pollinosis has shown large increases especially in city areas, rather than in the rural, with increases being remarkable especially among people of young age. In order to elucidate factors for sensitization and onset of Japanese cedar pollinosis, a questionnaire survey was conducted and serum IgE antibody levels examined in junior high school students living in three different areas, i.e., city, seaside and mountainous areas in Aichi prefecture. Prevalence rate of Japanese cedar pollinosis, estimated from the number of students having both nasal or ocular allergic symptoms and positive RAST, was the 30.1% in the mountainous area with high Japanese cedar pollen count, being greater than the 15.3% in the city area and the 5.8% in the seaside area, both with low pollen counts. Among the students showing high serum mite (Dermatophagoides farinae) antibody levels, greater positive rates for sensitization to Japanese cedar or orchard grass pollens was observed, compared to mite-unsensitized students. This result suggests that sensitization to seasonal antigens may be corresponding to the capability for sensitization to the mites. Besides this finding, the analysis of replies to the questionnaires showed that students living in concrete or steel frame buildings had greater serum levels of antibody to Japanese cedar pollen than those living in wood buildings, indicating a possible relationship between the pollen sensitization and indoor environments. PMID- 8704272 TI - [Depression symptoms among Chinese students in Japan]. AB - The mental health of foreigners in Japan, which shows a prominent increase in number recently was studied. A major group of these foreigners are Korean and Chinese, as their countries and Japan historically had a close relationship. The Chinese population has shown large increases, quadrupling over a period of 10 years. This population is characterized by purpose of residence; with most of them visiting Japan to study. Using the Beck Depression Inventory (BDI) self rating scale, we examined depression symptoms among two groups of Chinese students studying in Japan; 71 students of Mie university (MU) and 90 students of Japanese language schools (JLS) in Mie prefecture. BDI examination revealed that 28.9% (mild; 22.2%, moderate; 3.3%, severe; 3.3%) of Chinese JLS students and 23.9% (mild; 22.5%, severe; 1.4%) of Chinese MU students were depressed. Chinese JLS students showed significantly higher total BDI scores than Chinese MU students (p < 0.05). BDI scores of item D (lack of satisfaction), J (crying spells) and S (weight loss) were also significantly elevated in Chinese JLS students (D: p < 0.01, J: p < 0.05, S: p < 0.01). These results suggest that Chinese JLS students experience more stress than Chinese MU students. PMID- 8704273 TI - [Comparison of life habits and health examination data between smokers and ex smokers]. AB - Health practice and health examination data were compared between smokers and ex smokers in 6,076 male subjects, whose ages ranged from 40 to 59 years, who participated in health examinations in 1993 in 17 towns of Gunma Prefecture. A total of 4,629 (76.2%) subjects replied to the interview on health practice and also had health examination data that could be used. The numbers of smokers and ex-smokers were 2,681 and 245, respectively for a total of 2,926 target population. The percentage for those having breakfast daily was significantly higher in ex-smokers than in smokers in the age 40's group (p < 0.01). The percentage for those doing exercise more than one day a week was significantly higher in ex-smokers than in smokers in the age 50's group (p < 0.05). Mean body mass index (BMI) was significantly higher in ex-smokers than in smokers in the age 40's and 50's groups, with p < 0.05 and p < 0.01, respectively. Mean high density lipoprotein cholesterol (HDLC) was significantly higher in ex-smokers than in smokers in the age 40's group (p < 0.05). The two way analysis of variance on BMI and HDLC was conducted by smoking and drinking factors. BMI was related to only smoking in the age 40's and 50's groups. Smoking and drinking habit related to HDLC in the age 40's group and the latter habit had a relatively large association to HDLC. From the association between BMI and smoking habit, we speculate that exercise was practiced by ex-smokers due to the obesity potential of the ex-smoking status. PMID- 8704274 TI - [Medical services utilization of the elderly during the last six months of life]. AB - The purpose of this study was to describe the characteristics of medical services utilization during the last six months of life. Subjects were 150 elderly who lived in a community and died in 1990. By each subject's medical claims during the last six months of life, the number of hospital days, the number of visits for ambulatory care, the number of physicians' and nurses' visits for home care, medical expenditure, and the utilization of medical care activities were measured as utilization variables. Subjects were classified into four subgroups by the combination of cancer and cerebrovascular disease. These diseases have an important impact on medical services utilization during the last six months of life. Utilization variables of each subgroup were compared. The findings were as follows; 1. During the last six months of life, hospital stays of cerebrovascular disease patients were for long periods, while cancer patients frequently and repeatedly changed the place of care between hospital and home. 2. The number of visits for ambulatory care and the number of medical care providers utilized by cancer patients were greater than that of cerebrovascular disease patients. Cerebrovascular disease patients whose stay at the hospital was short made greater use of physicians' and nurses' visit for home care. 3. The medical expenditure of cancer patients was higher than that of cerebrovascular disease patients, although the number of hospital days of cancer patients was less. 4. Most of the cancer patients utilized transfusion and urological treatment. These results suggest that the utilization of medical care activities of cancer patients was more intensive than that of cerebrovascular disease patients. However cancer patients made little use of resuscitation procedures. PMID- 8704275 TI - Glucose-regulated stresses confer resistance to VP-16 in human cancer cells through a decreased expression of DNA topoisomerase II. AB - Glucose-regulated proteins (GRPs) are induced in cells by a variety of stress conditions such as treatment with 2-deoxyglucose, glucosamine, or the calcium ionophore A23187. We found that resistance to topoisomerase II (topo II) inhibitors, VP-16 and adriamycin, was induced by these treatments in human colon cancer HT-29 cells. Similar VP-16 resistance occurred in human ovarian cancer A2780 and breast cancer MCF-7 cells. The VP-16 resistance was reversible, since the sensitivity of the cells to VP-16 recovered within 24 h after the stresses were removed. Western blotting analysis showed that under these stress conditions the cellular contents of topo II alpha were decreased. The decreased expression of topo II was reversed to control levels within 24 h following removal of the stresses. The decrease in topo II levels under the stress conditions correlated well with the induction of GRP78 and 94. The close correlation between topo II and GRPs suggests that topo II is a protein sensitive to the glucose-regulated stresses. Since hypoxia and nutrient deprivation, which are also GRP-inducing conditions, could occur naturally in the solid tumors, the stress-associated cellular resistance through decrease in topo II levels may be a mechanism of the natural resistance of the solid tumors to topo II-directed chemotherapy. PMID- 8704276 TI - Granulocyte colony-stimulating factor receptor mediated cell growth and differentiation. AB - Granulocyte colony-stimulating factor (G-CSF) can elicit different biological responses through interaction with its receptor (G-CSFR). A series of G-CSFR expressing clones were established from WEHI-3B D+ myelomonocytic leukemia cells and interleukin-3 (IL-3) dependent Ba/F3 preB cells by transduction of a G-CSFR cDNA. The overexpression of the G-CSFR from the expression vector promoted differentiation in WEHI-3B D+ cells exposed to G-CSF, while, in contrast, the cytokine supported growth in Ba/F3 cells transfected with the expression plasmid in the absence of IL-3. Western blotting, immunoprecipitation and cell surface labeling with Na[125I] followed by immunoprecipitation with an anti-G-CSFR antibody revealed different patterns of detected proteins in these two cell lines, implying that different forms of the G-CSFR were generated which produced distinct biological responses and/or that different cell-type specific signal transducers were involved in the production of cellular differentiation or growth. PMID- 8704277 TI - Analysis of the interactions of SDZ PSC 833 ([3'-keto-Bmt1]-Val2]-Cyclosporine), a multidrug resistance modulator, with P-glycoprotein. AB - Multidrug resistance (MDR) is considered to be an important impediment to the effective treatment of cancer. P-glycoprotein, the drug efflux pump that mediates this resistance, can be inhibited by a wide variety of pharmacological agents, resulting in the circumvention of the MDR phenotype. SDZ PSC 833 ([3'-keto-Bmt1] Val2]-cyclosporine), a nonimmunosuppressive cyclosporine D derivative, was identified to be a potent MDR modulator (Gaveriaux et al. J. Cell Pharmacol. 2:225-234; 1991). In this study, the interactions of P-glycoprotein with two cyclosporine derivatives, SDZ PSC 833 and cyclosporine A (CsA, Sandimmune), were analyzed. SDZ PSC 833 enhanced the sensitivity of the MDR cells to anticancer drugs by increasing the accumulation and inhibiting the efflux of cytotoxic agents from resistant cells more efficiently than CsA. The two cyclosporine analogs competed with the labeling of P-glycoprotein by a photoactive cyclosporine derivative. In addition, membrane vesicles derived from resistant cells bound SDZ PSC 833. However, CsA was transported by P-glycoprotein, whereas SDZ PSC 833 was not actively transported. This resulted in a prolonged inhibitory effect by SDZ PSC 833. The studies suggest that the binding of SDZ PSC 833 to P glycoprotein in the absence of its transport from MDR cells mediated its high potency as an MDR reversing agent. In addition, the comparison of the two cyclosporine analogs indicated that limited chemical modifications of MDR reversing agents can affect their potential to inhibit P-glycoprotein function. PMID- 8704278 TI - Improved antitumor activity of cis-Bis-neodecanoato-trans-R,R-1,2- diaminocyclohexaneplatinum (II) entrapped in long-circulating liposomes. AB - Cis-bis-neodecanoato-trans-R, R-1,2-diaminocyclohexaneplatinum (II) (NDDP) has previously been formulated in conventional liposomes and shown to be nonnephrotoxic in humans, not cross-resistant with cisplatin in different in vitro and in vivo systems, and more active than cisplatin against murine models of experimental liver metastasis. The activity was attributed to the avid uptake of liposomes by the liver. To extend to the treatment of solid tumors outside the liver, NDDP was formulated, in this study, in long-circulating liposomes composed of egg phosphatidylcholine (PC), cholesterol (Chol) and polyethyleneglycol conjugated to phosphatidylethanolamine (PEG-PE). In vitro, PC/Chol/NDDP liposomes were barely toxic to murine melanoma cells B16-F0 (IC50 = 195.6 microM) as evaluated by tetrazolium dye colorimetric assay. Inclusion of PEG3000-PE into PC/Chol/NDDP liposomes significantly enhanced their cytotoxicity to a level that was comparable to that of DMPC/DMPG/NDDP liposomes (7.3 vs. 7.9 microM). Biodistribution study indicated that PC/Chol/PEG3000-PE/NDDP liposomes could preferentially localize in s.c. melanoma in C57BL/6 mice. PC/Chol/PEG3000-PE/NDDP liposomes were much more efficient in inhibiting tumor growth in vivo than free cisplatin, free NDDP or NDDP formulated in liposomes of other lipid composition. Compared with cisplatin, the in vivo toxicities of PC/Chol/PEG3000-PE/NDDP liposomes were also significantly reduced. Furthermore, if the tumor was treated with local hyperthermia after injection of PC/Chol/PEG3000-PE/NDDP liposomes, the tumor uptake of liposomes increased by 60%. The tumor inhibitory effect of PC/Chol/PEG3000-PE/NDDP was also significantly improved when combined with local hyperthermia. PMID- 8704280 TI - Reference listings in cancer research. PMID- 8704279 TI - P-glycoprotein expression in ovarian cancer cell line following treatment with cisplatin. AB - Human ovarian cancer cell line SKOV3 was grown during a period of four months in the presence of increasing concentrations of cisplatin (25-100 ng/ml). In the course of this treatment, the cells exhibited dramatic changes in morphology, including reduction in cell size, loss of cellular projections and clustering. This was accompanied by the appearance of P-glycoprotein (Pgp) on the cell membrane, as detected by flow cytometry and immunochemistry methods using the anti-Pgp monoclonal antibodies MRK16 and C219. The new cell line, designated SKOV3/CIS, was also resistant to alkylating agents, such as chlorambucil, similarly to the parental SKOV3 cells. In addition, it also acquired resistance to classical multidrug resistance drugs, such as doxorubicin, taxol and actinomycin D. Verapamil enhanced the sensitivity of SKOV3/CIS to doxorubicin (260-fold), in conformity with the proposed mechanism of Pgp in multidrug resistance (MDR), but it did not potentiate cisplatin cytotoxicity in SKOV3/CIS cells. Our results suggest that cisplatin can cause Pgp expression, and that both cisplatin-resistance and Pgp-mediated MDR phenotypes can coexist in some tumor types. Although Pgp does not appear to be responsible for cisplatin resistance, exposure to cisplatin can lead to the development of MDR phenotype, a complication that should be considered in clinical situations, especially in the chemotherapy of ovarian cancer. PMID- 8704281 TI - Microsatellite instability and loss of heterozygosity at the hMLH1 locus on chromosome 3p21 occur in a subset of nonsmall cell lung carcinomas. AB - Microsatellite alterations observed in tumor specimens may reflect genomic instability due to defective mismatch repair genes. To investigate whether this occurs in human nonsmall cell lung carcinomas we have analyzed microsatellite instability at 5 marker loci and loss of heterozygosity (LOH) at the hMLH1 locus on chromosome 3p21 using the polymerase chain reaction. Of a total of 49 nonsmall cell lung carcinomas examined, 43% (13 of 30 informative cases) showed LOH at 3p21 and 29% (14 of 49) exhibited microsatellite instability at one or multiple loci. LOH of the mismatch repair gene hMLH1 at 3p21 occurred in 82% (9 of 11 informative cases) of the tumors with microsatellite instability. This suggests that defects in the mismatch repair gene hMLH1 at 3p21 may be involved in microsatellite instability and tumorigenesis of a subset of nonsmall cell lung carcinomas. However, in those nonsmall cell lung carcinomas without microsatellite instability LOH at 3p21 probably involved another tumor suppressor gene(s) in this chromosomal region. PMID- 8704282 TI - Gonadotropin beta-subunit nicking enzyme (GBNE), a potential marker of early malignancies. AB - GBNE is an arginine-specific metalloprotease that cleaves human chorionic gonadotropin free beta-subunit in pregnancy serum. We tested GBNE activity in 539 serum samples, from individuals that were healthy, with benign diseases, or with a broad mixture of cancers. The mean GBNE activity in 130 samples from healthy individuals, 151 from those with benign disease, and 258 from cancer patients was 5.4 +/- 0.32, 5.8 +/- 0.20 and 16 +/- 1.2 units, respectively. ROC analysis indicated 86% discrimination between control samples and cancer. A cut-off of 12 units was selected. This was equaled or exceeded by 3.1% of samples from healthy individuals, 3.9% from those with benign disease, and 57% from those with cancer. There was no significant difference in detection of the different cancer primaries, breast (sensitivity 56%), gastrointestinal (53%), genitourinary (62%), gynecological (58%) and lung (55%) cancers. Sensitivity was highest for early stage, and lowest for advanced malignancies, for all cancer primaries. Taken together, sensitivity was 70, 44, 43 and 24%, for stages I, II, III and IV, respectively. GBNE is suggested to be a nonorgan site-restricted tumor marker with high sensitivity for early stage cancers. PMID- 8704283 TI - Elevations of DNA topoisomerase I catalytic activity and immunoprotein in human malignancies. AB - DNA topoisomerase I (topo I) is the molecular target for the camptothecin group of anticancer drugs. These drugs are showing activity against a wide array of human tumors. Many data have indicated that the sensitivity of a tumor cell to the camptothecins is dependent on tumor topo I levels. Drug-sensitive cells have high levels of topo I. Unfortunately, there is still a relative lack of information on topo I levels in human malignancies. Because of this, we investigated topo I activity and immunoprotein levels in a variety of normal murine and human tissues, as well as tissues obtained from several carcinomas, lymphomas, and sarcomas. Flow cytometric analysis was also performed on the neoplastic specimens to determine the percentage of cycling cells. Topo I catalytic activity was detected in all normal tissues at a fairly constant level. The average topo I catalytic activity in normal mammalian tissues was 2.7 +/- 1.3 x 10(4) units/mg protein (range 1.1 to 5.0 x 10(4)). Topo I catalytic activity was much more variable in human malignancies and ranged from a low of 1.4 x 10(4) units/mg protein in a rhabdomyosarcoma to a high of 160 x 10(4) units/mg protein in a poorly differentiated ovarian carcinoma. Western blot analysis with either a mouse monoclonal antibody or scleroderma antibodies directed against topo I revealed that the elevated topo I catalytic activity levels in the malignant tissues are due to elevated amounts of topo I immunoprotein. It is possible that the high topo I levels that characterize several different types of human malignancies might indicate that these tumors would be sensitive to many of the new drugs that target topo I. PMID- 8704284 TI - Energy metabolism of adriamycin-sensitive and -resistant Ehrlich ascites tumor cells. AB - Respiration, glycolysis, utilization of carbon from 14C-labeled glucose and the activities of some regulatory enzymes of the Krebs cycle and glycolysis in adriamycin-sensitive (EH-WT) and -resistant (EH-ADR) Ehrlich ascites tumor cells have been investigated. The following summarizes the results: 1. Compared with wild-type cells, EH-ADR cells exhibited an enhanced rate of oxygen consumption as well as of ATP production (2-fold). 2. When the cells were supplied with glucose as the only added energy source, the aerobic lactate production was 30% higher in EH-ADR cells. However, in spite of the enhanced glycolysis, 50% of total cell ATP was still supplied by oxidative phosphorylation, whereas in EH-WT cells 65% of ATP was derived from glycolysis. 3. The activities of the regulatory enzymes were remarkably more elevated in EH-ADR cells. 4. The amount of glucose carbon atoms metabolized through the Krebs cycle and pentose phosphate pathway in EH-ADR cells was significantly higher than in EH-WT cells. 5. These differences confirmed a modified energy metabolism in resistant cells and reflected metabolic adaptations associated with the development of multidrug resistance. PMID- 8704285 TI - Reference listings in cancer research. PMID- 8704286 TI - Cisplatin-induced alterations in the expression of the mRNAs for UV-damage recognition protein. AB - Enhanced DNA repair is believed to be an important mechanism of the cisplatin resistant phenotype. UV-damage recognition protein (UV-DRP) recognizes and binds to DNA lesions and may play a role in DNA nucleotide excision repair and/or replicative bypass (which is associated with post-replication repair). Potential alternations in the expression of mRNAs for UV-DRP were analyzed in this study. Two pairs of parental and cisplatin-resistant human ovarian carcinoma cell lines were utilized. Gene expression level was assessed by northern blot hybridization. No alterations in mRNA levels for the large subunit of UV-DRP were found following cisplatin treatment, whereas mRNA levels for the small subunit of UV DRP were induced up to 4.5-fold. The time-course and concentration-response of this induction corresponded to the previously reported increase in the UV-DRP binding activity, as measured by gel shift assay. UV-DRP binding activity in cell extracts corresponds to expression of small subunit mRNA but not to expression of large subunit mRNA. These data suggest that the small subunit may be limiting for UV-DRP activity. PMID- 8704287 TI - Phase-contrast magnetic resonance angiography for the determination of cerebrovascular reserve. AB - Cerebrovascular reserve (CVR) is the potential for cerebral arteriolar dilatation to occur, in response to decreased cerebral perfusion pressure, in order to maintain constant cerebral blood flow. Diminution or absence of CVR is considered a risk factor for stroke. Current methods for determining CVR include single photon emission computed tomography, positron emission tomography and transcranial Doppler (TCD) ultrasonography. However, significant advantages could derive from the utilization of magnetic resonance angiography (MRA) based on the concurrent acquisition of hemodynamic information (CVR and collateral flow) with phase-contrast (PC) techniques and vascular morphology with three-dimensional, time-of-flight methods. With a 1.5-T scanner and acetazolamide (AZM), an arteriolar dilator, CVR was determined in 7 normal subjects. Mean flow velocity in the middle cerebral arteries was determined by PC MRA before and after AZM administration. For comparative purposes, mean flow velocities in the same middle cerebral arteries were determined by TCD before and after AZM administration. The mean flow velocities were as follows (mean +/- standard deviation, n = 7): 40 +/- 8 (PC MRA) versus 61 +/- 10 cm/sec (TCD) before AZM treatment and 58 +/- 11 (PC MRA) versus 85 +/- 15 cm/sec (TCD) after AZM administration. The increase in mean flow velocity (before vs after AZM), that is, the CVR, was 45 +/- 11% as shown by PC MRA and 39 +/- 14% as shown by TCD. Although significant differences were present between the mean flow velocities measured before and those after AZM administration, as determined by PC MRA and TCD, the CVR was not significantly different (45 vs 39%, respectively). These preliminary results suggest that PC MRA may be a method for determining CVR. PMID- 8704288 TI - Time course of the acetazolamide effect in patients with extracranial carotid artery disease. AB - The acetazolamide test of cerebrovascular reserve is performed as a two-point transcranial Doppler (TCD) measurement of the flow velocity in the middle cerebral artery (MCA) before and 15 to 20 minutes after injection of the carbonic anhydrase inhibitor. The time course was evaluated by bilateral TCD monitoring over 45 minutes in 51 patients with extracranial carotid artery disease. Twenty nine patients in Group 1 showed a unilateral and 22 patients in Group 2, a bilateral stenosis of the internal carotid artery (ICA). Thirty-one normal hemispheres in 18 normal subjects provided baseline control values. The most useful parameter for the description of the time course was the integral of the mean flow velocity. In both groups integral of the mean flow velocity was significantly reduced compared to that in normal persons (Group 1: 587.2 +/- 538.3 cm, p < 0.01; Group 2: 728.9 +/- 397.1 cm, p < 0.01; normal persons: 967.8 +/- 350.0 cm). The maximal increase of mean flow velocity in a two-point measurement also was significantly lower in the patient groups, but did not reach the same level of significance in Group 2 (Group 1: 19.6 +/- 12.3 cm/sec, p < 0.01; Group 2: 24.2 +/- 10.4 cm/sec, p < 0.05). Thirty-four of 73 hemispheres had reduced integral of the mean flow velocity, but only 29 had a reduced maximal increase. These data indicate that continuous monitoring and evaluation of the time course of the acetazolamide effect provide additional information on cerebrovascular reserve and help to detect an increased number of patients with impaired cerebrovascular reserve. PMID- 8704289 TI - Clinical significance of increased uptake of HMPAO on brain SPECT scans in acute stroke. AB - Single-photon emission computed tomography (SPECT) with 99mtechnetium hexamethylpropylamineoxime (HMPAO) noninvasively shows brain perfusion in patients after acute stroke. However, the clinical significance of the increased HMPAO uptake remains unclear. In this study, consecutive patients with hemispheric hemorrhagic and ischemic stroke admitted to the hospital were evaluated prospectively. The increased uptake of HMPAO was determined by visual analysis of SPECT images. The pathogenic mechanism of ischemic stroke was determined using the clinical and computed tomography (CT) criteria including the Toronto Embolic Scale. Of the 500 consecutive patients with acute hemispheric stroke, SPECT was performed in 458 at a mean time 5 +/- 7 days after the onset of symptoms. A strong association was found between SPECT perfusion patterns and pathogenic subtypes of stroke (p < 0.0001). Thus, in 95% of patients with intracerebral hemorrhage the focal absence of perfusion was found, and 26% of lacunar infarctions presented with a normal SPECT appearance. The mean volume of lacunar lesions that did not produce significant abnormalities on SPECT was 2.5 +/- 1.2 ml. Increased HMPAO uptake was associated with a cardioembolic mechanism of stroke: High and mixed perfusion patterns were present subacutely in 29% of patients with cardioembolic stroke, compared to 15% of patients with other types of ischemic stroke (p < or = 0.0006). The increased uptake of HMPAO on SPECT as determined by visual analysis is associated with a cardioembolic mechanism of cerebral ischemia, which could be explained by glutathione-mediated trapping of the tracer during reperfusion and later in newly developed granulation tissue. HMPAO-SPECT may help in early management decisions since it indicates stroke pathogenesis and evolution. PMID- 8704290 TI - Functional imaging predicts cognitive decline in Alzheimer's disease. AB - Eighteen patients with Alzheimer's disease were studied with positron emission tomographic measurements of regional cerebral metabolism of glucose. All patients were initially diagnosed and evaluated, underwent positron emission tomography, and then were followed with annual reevaluations, at which time the Mini-Mental State Examination (MMSE) was performed. Patients were followed for an average of 2.5 years, and the rate of cognitive decline was calculated by determining the rate of change in the MMSE score defined as the MMSE score at the initial evaluation minus the MMSE score at the last examination, divided by the number of months between testing. The regional cerebral metabolic rates for glucose determined at the time of the first MMSE were then regressed on these changes in scores. Results showed that glucose metabolic rates in posterior temporal and primary visual cortex regions were significantly correlated with the subsequent rate of cognitive deterioration. These associations were not confounded by age, length of follow-up, baseline MMSE score, or education. Stratification on gender suggested that these associations were much stronger in women than in men. These results replicate previous findings showing that functional brain imaging is predictive of the rate of cognitive decline in Alzheimer's disease. PMID- 8704291 TI - Neuroimaging characteristics in carbon monoxide toxicity. AB - Neuroimaging studies for 18 patients carrying carbon monoxide toxicity as a discharge diagnosis were reviewed. The most common positive findings were low density lesions in the globus pallidus (7/18, 39%) and deep white matter changes (5/18, 28%). Six computed tomography head scans showed no acute changes. Advanced age, method of exposure (intentional vs accidental), and severity of carboxyhemoglobin level did not predict neuroradiological or clinical outcomes. Early neuroimaging could not be used to predict clinical courses (death or coma vs discharge to either an institution or a home) in the patient population studied. PMID- 8704292 TI - The spectrum of neuroimaging abnormalities in solvent abuse and their clinical correlation. AB - The brain magnetic resonance (MR) images and medical records of 6 patients with a history of chronic toluene abuse were reviewed retrospectively. The imaging findings were correlated with clinical impairment. The major findings consisted of atrophy (6 patients), white matter T2 hyperintensity (6 patients), and T2 hypointensity involving the basal ganglia and thalami (5 patients). Also seen was focal enhancement, previously unreported for patients who abused toluene. This study showed a correlation between the degree of neurological impairment and extent of white matter disease. There was no correlation between the severity of white matter changes on MR images and the presence of T2 hypointensity or duration of toluene abuse. There was no definite clinical evidence of damage to the basal ganglia and thalami despite the MR imaging finding of T2 hypointensity. Temporal evidence against the theory that toluene accumulation causes the T2 hypointensity was found. PMID- 8704293 TI - Serial transcranial Doppler monitoring after transient ischemic attack. AB - The natural course of ultrasonically detectable microembolism in patients with a symptomatic extracranial atherosclerotic lesion is not completely understood. Furthermore, the potential impact of therapeutic management on microemboli detection is anecdotal. A 58-year-old man who experienced a left-hemisphere transient ischemic attack presented with an extracranial ipsilateral high-grade carotid stenosis. He was studied 12 times in 12 months during different medical management, during which time he was symptom free, even though transcranial Doppler evidence of microemboli continued for 5 months. PMID- 8704294 TI - Magnetic resonance imaging of petrous carotid aneurysms. AB - Aneurysms arising from the petrous segment of the internal carotid artery are rare and the majority of reports are based on evaluations with x-ray catheter angiography (XRA) and computed tomography (CT). Due to the paucity of magnetic resonance imaging (MRI) information, the author describes 2 patients with petrous carotid aneurysms evaluated with MRI and magnetic resonance angiography (MRA). Compared to XRA and CT, MRI and MRA are less invasive and provide more information regarding the aneurysm, including the size, shape, and site; the absence or presence and extent of hemorrhage, thrombus, and bony erosion; patency of the vessel; neck; and the extent of perianeurysmal compromise of vessels, cranial nerves, and parenchyma. PMID- 8704296 TI - The evaluation of symptomatic carotid artery disease. A survey of specialty practices in Rhode Island. AB - Severe, symptomatic carotid artery disease is most often diagnosed by conventional cerebral angiography. Noninvasive tests are commonly used to identify candidates for angiography and endarterectomy. The purpose of this study, a mail and telephone survey of academic and community neurologists, neurosurgeons, and vascular surgeons in Rhode Island in 1994, was to determine which noninvasive tests physicians used to evaluate these patients and how the test results were used to select patients for angiography. One hundred (86%) of a possible 116 responses were collected. Seventy-six percent of physicians chose carotid duplex ultrasound as the first diagnostic test, a percentage significantly higher than that for any other test (p < 0.0001). Fourteen percent chose angiography without a prior screening test; 3% chose magnetic resonance angiography (MRA) prior to angiography, 6% chose carotid duplex ultrasound plus MRA prior to angiography, and 1% chose MRA without angiography. The specific noninvasive test results required for angiography referral were surprisingly variable, both within and across the three physician specialties. Vascular surgeons and neurosurgeons were more likely to choose angiography without prior screening tests than were neurologists. In conclusion, the majority of specialty physicians in the state of Rhode Island chose carotid ultrasound as the screening test of choice for the evaluation of a potential endarterectomy candidate. Few responding physicians chose MRA in combination with carotid duplex ultrasound prior to angiography, or MRA prior to angiography or MRA alone. Awareness of the actual practice patterns of specialists may lead to programs of education for physicians to improve practice and patient outcome through more refined use of diagnostic tests prior to angiography or endarterectomy. PMID- 8704295 TI - Size of the corpus callosum in cerebral palsy. AB - It has been suggested that the size of the corpus callosum may have diagnostic significance in cerebral palsy, although this relationship is incompletely defined. Ninety-one patients with cerebral palsy had been studied by magnetic resonance imaging in the 5-year period from 1990 to 1994. Fifty-seven of these 91 patients had a technically appropriate midsagittal magnetic resonance image for quantitative morphometric analysis. The ratio of the area of the corpus callosum to the area of the supratentorial brain was compared to published age- and gender specific norms. Imaging findings were correlated with clinical history and cause of cerebral palsy. The corpus callosum was of normal size in 43 patients and more than 2 standard deviations below the mean in 14 patients. The causes for cerebral palsy included hypoxic ischemic encephalopathy (32), cerebral dysgenesis (8), and porencephalic strokes (6); the etiology could not be established in 11 patients. The size of the corpus callosum was highly correlated with the cause of cerebral palsy, such that all patients with cerebral dysgenesis had hypoplasia of the corpus callosum (one-sided z test, p < 0.0001). Conversely, the callosum was of normal size in 32 of 38 patients with hypoxic ischemic encephalopathy and porencephalic strokes. The presence of a hypoplastic corpus callosum is highly associated with cerebral dysgenesis as a cause for cerebral palsy. PMID- 8704297 TI - "Semantic" conduction aphasia from a posterior insular cortex infarction. AB - A unique infarction limited to the posterior insula and intrasylvian parietal opercular cortex produced a subtype of conduction aphasia, characterized by a predominance of semantic paraphasias. Temporal lobe hypoperfusion seen on hexamethylpropyleneamineoxime single-photon emission computed tomography in the absence of any signs of ischemia suggested that cortical diaschisis played a role in the emergence of this syndrome. PMID- 8704298 TI - Adult-onset complex partial seizures as the presenting sign in colpocephaly: MRI and PET correlates. AB - In a 30-year-old man, adult-onset complex partial seizures were associated with colpocephaly as evident from magnetic resonance images. The structurally normal lateral neocortex of the right temporal lobe showed a severe reduction in regional glucose consumption corresponding to the epileptic focus as judged from the surface electroencephalogram. In contrast to previously reported patients, this patient had no focal neurological signs, and neuropsychological testing revealed normal general intelligence. Thus, this patient adds a benign variant to the clinical spectrum of this cerebral developmental disorder. PMID- 8704299 TI - Pseudoprogression of cerebral cavernous angiomas: the importance of proper magnetic resonance imaging technique. AB - Magnetic resonance imaging (MRI) is the method of choice to detect cerebral cavernous angiomas. Specifically tailored MRI studies are necessary to evaluate patients with cerebral cavernous angiomas. The patient described here, for whom serial MRIs performed on different scanners falsely suggested rapid disease progression, illustrates the point. PMID- 8704300 TI - Enlarged Virchow-Robin spaces along the medullary perforators in a child with seizures. AB - Enlarged Virchow-Robin spaces along the medullary perforators on brain magnetic resonance images (MRIs) of a 4-year-old, neurologically intact child with seizures are described. The differential diagnosis of cystic spaces in the centrum semiovale is discussed. The sparse MRI literature on bright signal intensities in childhood is reviewed. PMID- 8704301 TI - Distinctive serial magnetic resonance changes in a young woman with rapidly evolved neurocysticercosis, with positron emission tomography results. AB - A woman aged 21 years noticed numbness of the left hand, dysarthria, and right hemiparesis. Magnetic resonance imaging (MRI) showed homogeneously enhanced lesions in the bilateral cerebral white matter; these enlarged within 2 weeks, showing ring-like enhancement, and then spontaneously decreased in size and enhancement. Neurocysticercosis was confirmed by repeated, significantly positive serology results and the dramatic response to praziquantel. Subsequently, positron emission tomography (PET) revealed persisting regions of possibly decreased cerebral uptake of [18F]2-fluoro-2-deoxyglucose. This is the first documentation of such distinctive serial MRI changes and the results of PET in neurocysticercosis. PMID- 8704302 TI - Bilateral thalamic infarctions can sometimes accompany midbrain infarction and rarely accompany pontine infarction. PMID- 8704303 TI - Inhibitory effects of green tea polyphenols on growth and cellular adherence of an oral bacterium, Porphyromonas gingivalis. AB - Effects of polyphenolic compounds isolated from green tea (Camellia sinensis) on the growth and adherence of Porphyromonas gingivalis onto human buccal epithelial cells were investigated. Green tea polyphenols, especially (-)-epigallocatechin gallate (EGCg) which is a dominant component of tea polyphenols, completely inhibited the growth and adherence of P. gingivalis onto the buccal epithelial cells at concentrations of 250-500 micrograms/ml. Among the polyphenolic compounds, (-)-epicatechin gallate (ECg) and (-)-gallocatechin gallate (GCg) were effective next to EGCg in these activities. On the other hand, (+)-catechin (C(+)), (-)-epicatechin (EC), (+)-gallocatechin (GC), and (-)-epigallocatechin (EGC) had very much less activity. These results indicate that the inhibitory effect on the adherence of P. gingivalis onto the buccal epithelial cells is attributed to the presence of the galloyl moiety, which is ester-linked with the 3-OH of the catechin moiety in the polyphenolic compounds. PMID- 8704305 TI - A method to invert DNA segments of the Bacillus subtilis 168 genome by recombination between two homologous sequences. AB - We developed a method that allows rapid isolation of mutant Bacillus subtilis 168 carrying an inversion of a specific DNA segment of the genome. Two incomplete neomycin resistance gene cassettes were integrated at both ends of the 1652-kb segment to be inverted. Reciprocal recombination within the 590-bp homologous region of these two cassettes created an intact neomycin resistance gene with concomitant inversion of the 1652-kb segment flanked by the two cassettes. Structure of the mutant genome was verified by analyzing the physical map for rare cutting endonucleases, SfiI, NotI, I-CeuI, and I-SceI. The inversion rate was estimated to be 6.9 +/- 1.4 x 10(-8)/cell/cell division at 37 degrees C. The method should be in principle applicable not only to other regions of the B, subtilis genome but also to other bacterial genomes. PMID- 8704304 TI - Protective effects of chlorogenic acid on paraquat-induced oxidative stress in rats. AB - The protective effects of chlorogenic acid on paraquat-induced oxidative stress were examined in rats. The activities of erythrocytes and liver glutathione peroxidase, and of both liver catalase and glutathione reductase, which were increased by feeding paraquat, declined to the levels in the control rats by supplementing chlorogenic acid to the paraquat diet. The activity of superoxide dismutase was not changed by dietary paraquat or by supplementing chlorogenic acid to the paraquat diet. Paraquat in the diet markedly decreased the liver triacylglycerol and phospholipid concentrations, as well as the food intake and body weight gain, while chlorogenic acid protected against these decreases. These in vivo results and the in vitro superoxide anion scavenging activity of chlorogenic acid suggest that chlorogenic acid acted preventively against paraquat-induced oxidative stress. PMID- 8704306 TI - Flavonoids as inhibitors of NADP-malic enzyme and PEP carboxylase from C4 plants. AB - The inhibitory effects of flavonoids on the activity of two photosynthetic enzymes such as phosphoenolpyruvate carboxylase (PEPCase) and NADP-dependent malic enzyme (NADP-ME) were evaluated. The glycosylation of hydroxyl groups on the flavonoids resulted in compounds that behaved as gradually weaker inhibitors with increased size of the substituent. Quercetin and baicalein showed a competitive inhibition pattern vs. NADP+ for NADP-ME, and a similar model for both flavonoids vs. phosphoenolpyruvate (PEP) was observed when tested on PEPCase. K(i) for NADP-ME inhibition at pH 7.0 were 0.83 microM and 1.54 microM for quercetin and baicalein, respectively. K(i) for PEPCase inhibition were 0.17 microM and 0.79 microM (quercetin and baicalein, respectively), indicating that these compounds are the most potent inhibitors described for this carboxylase. I50 values for these and other flavonoids were in the micromolar range. A tentative physiological role for the inhibitory effects observed on PEPCase is discussed. PMID- 8704307 TI - Cloning and characterization of mycovirus double-stranded RNA from the plant pathogenic fungus, Fusarium solani f. sp. robiniae. AB - A mycovirus (named FusoV) from the phytopathogenic fungus, Fusarium solani f. sp. robiniae SUF704, has two kinds of double-stranded (ds) RNA genomes, designated M1 and M2. The cDNAs were constructed from FusoV genomic dsRNAs. The sequences of M1 and M2 cDNAs comprised 1645 and 1445bp, respectively. Sequence analysis showed that each dsRNA had a single long open reading frame (ORF) on only one of the strands. M1 ORF encodes a 519-amino acid residue polypeptide with a predicted molecular mass of 60 kDa. RNA-dependent RNA polymerase-conserved motifs were identified in the predicted amino acid sequence, and the polymerase synthesized dsRNA in vitro. The M2 ORF encodes a polypeptide of 413 amino acid residues with a predicted molecular mass of 44 kDa. The predicted amino acid sequence contained the sequence corresponding to those found in the purified 44-kDa capsid protein of FusoV. PMID- 8704309 TI - Isolation and genetic characterization of pGKL killer-insensitive mutants (iki) from Saccharomyces cerevisiae. AB - The linear double stranded DNA plasmid pGKL1 encodes the yeast killer toxin complex (Gunge et al., 1981) of which the killing mechanism is not understood. We isolated and characterized eight mutants in Saccharomyces cerevisiae that were insensitive to both the intracellularly expressed 28-kDa killer subunit and the native killer toxin complex. These mutations (iki1 through iki5) were all recessive, and classified into five complementation groups. The iki2 mutation was mapped to a position near the centromere on chromosome XIII. We developed a novel screening system to isolate the DNA fragments complementing the iki mutations from a Saccharomyces gene library, and isolated three DNA fragments that complement the iki1, iki3, and iki4 mutations, respectively. PMID- 8704308 TI - Immunosuppressive activity of streptonigrin in vitro and in vivo. AB - The immune system is composed of various cells with distinct functions. Thus, highly selective immunomodulators are necessary for artificial regulation of immune reactions. We screened microbial products for such immunomodulators and we identified streptonigrin as a selective suppressor of B-cell proliferation induced by lipopolysaccharide. Streptonigrin directly suppressed the late phase of proliferation of B-cells. The inhibition of topoisomerase II was implicated as the mechanism of the B-cell-selective suppression. In cultured cell lines, however, streptonigrin preferentially suppressed the growth of an interleukin-3 dependent myeloid cell line rather than B-cell lines. In addition, the treatment with streptonigrin in vivo suppressed T-cells more significantly than B-cells and dramatically reduced the spleen weight. These results suggest that streptonigrin preferentially suppresses myeloid T-cell precursors in vivo. PMID- 8704310 TI - Rate parameter changes by added albumin in the microsomal oxidative demethylation of deuteriated and non-deuteriated 4-methoxyanisole. AB - Bovine serum albumin (BSA) added to the reaction medium for the oxidative demethylation of 4-methoxyanisole and its "di-CD3" isotopomer ([d6]methoxyanisole), when catalyzed by liver microsomes from untreated rats, decreased the Km values and increased the V(max)/Km (= V/K) values. The Vmax values were not markedly altered. The values for the deuterium isotope effect on V(max) and V/K for the reaction with this isotopomer were between 2.2 and 2.8, and that on Km was close to unity. The magnitude of the isotope effect was not significantly changed by adding BSA. The intramolecular isotope effect with [mono CD3]4-methoxyanisole ([d3]methoxyanisole) in liver microsomes from untreated rats was between 10.3 and 10.8, which was not significantly changed by BSA. Liver microsomes from rats treated with phenobarbital resulted in the intramolecular isotope effect value in the absence of BSA being between 7.2 and 9.1, which was not significantly altered by BSA. Based on these data, the calculated apparent rate constant for the enzyme-substrate complex formation was markedly increased by up to about 1.9- and 3.5-fold by 1% and 2% of BSA added, respectively. PMID- 8704311 TI - Production of recombinant human monoclonal antibody using ras-amplified BHK-21 cells in a protein-free medium. AB - A ras oncogene-amplified recombinant BHK-21 cell line (ras-rBHK-IgG) has been established, and was shown to hyperproduce the recombinant IgG chimeric human monoclonal antibody (hMAb) AE6F4, which recognizes lung cancer cells. We found that the ras-rBHK-IgG cell could be easily cultured in a protein-free ERDF medium supplemented with iron(III) nitrate, hydroxyethyliminodiacetic acid, and non protein synthetic attachment factor as well as in a serum-free ERDF medium supplemented with insulin, transferrin, ethanolamine, and sodium selenite. The productivity of recombinant hMAb from the cells cultured in dishes at high cell densities was higher in protein-free medium than in serum-containing medium. True high density culture of the ras-rBHK-IgG cells was done in protein-free medium using the Tecnomouse, which is a novel hollow fiber bioreactor system. After culture for 30 days in protein-free culture, a total amount of about 14 mg of the recombinant hMAb AE6F4 was obtained, and was shown to be reactive against lung cancer cells in tissues. PMID- 8704312 TI - Should Petasospora BOIDIN et ABADIE (Saccharomycetaceae) be retained?--the phylogeny based on the partial sequences of 18S and 26S ribosomal RNAs. AB - The six strains of the Pichia species, once classified in the genus Petasospora, were examined for their 18S (positions 1451-1618, 168 bases) and 26S (positions 1611-1835, 225 bases and 493-622, 130 bases) rRNA partial base sequencings. In the 18S rRNA partial base sequencings, the type species of the genus Petasospora (Petasospora rhodanensis) was found to be closely related to Pichia anomala (identical to Hansenula anomala, type species of genus Hansenula) with base differences of two, but not to Pichia membranaefaciens, the type species of the genus Pichia, with base differences of ten. However, the species was not so closely related to P. anomala in the 26S rRNA partial base sequencings with base differences of twelve and 68 percent similarity. The genus Petasospora was known to be a very heterogeneous taxon phylogenetically with base differences of thirty one to three and ninety-two to eight and with 40 to 83 percent similarities. The sequence data obtained here and the phenotypic features described previously indicate that the type species of the genus Petasospora (Pet. rhodanensis, identical to Pichia rhodanensis) is adequate to be accommodated temporarily in the genus Pichia, a heterogeneous taxon, until the precise taxonomic position of the type species is defined. PMID- 8704313 TI - Polymerization degree of oligomethionine to determine its bioavailability when added to a low-protein diets. AB - Oligo-L-methionine ethylester (OMOEt) prepared by the papain-catalyzed oligomerization of L-methionine ethylester (MetOEt) is a mixture of pentamer to dodecamer and has nearly the same supplementary effect as free methionine (Met) for the growth of rats when added to a low casein diet, but its supplementary effect to a low-soy protein isolate (SPI) diet is not consistent and depends on the degree of polymerization. Rats were fed for 2 wk with an 8% casein or 10% SPI diet supplemented with 0.3% L-Met, each chemically synthesized MetnOEt with a polymerization degree (n) of 6, 7, 8, or 9, or with OMOEt prepared by papain catalyzed polymerization of MetOEt. Met6OEt, Met7OEt, and Met8OEt had nearly the same supplementary effect on the growth of rats, as did free Met, both with the 8% casein and 10% SPI diets. The supplementary effect of Met9OEt was not significantly lower than that of Met when added to the 8% casein diet, but was significantly lower when added to the 10% SPI diet. The digestibility of Met9OEt supplemented to the 8% casein and 10% SPI diets was 50.5% and 35.6%, respectively. It appears likely that there is a gap in the bioavailability of oligomethionine between the octamer and nonamer when added to a low-protein diet, probably due to the rigidity of the structure increasing with the polymerization degree by alpha-helix formation. Although the differences in absorption rate of Met from OMOEt for a short time after feeding has been related to the different effects of supplemented OMOEt, the absorption rate of OMOEt for 30 min after feeding was not considered to be the main cause of the differential effects of OMOEt in this experiment. PMID- 8704314 TI - Purification and characterization of a thermostable alkaline protease from Thermoactinomyces sp. E79 and the DNA sequence of the encoding gene. AB - A thermophilic Thermoactinomyces sp. E79 producing a highly thermostable alkaline protease was isolated from soil. The protease, produced extracellularly by Thermoactinomyces sp. E79, was purified by DEAE-Sepharose CL-6B and Butyl Toyopearl 650M column chromatography. The relative molecular mass was estimated to be 31,000 by SDS-polyacrylamide gel electrophoresis. Enzyme activity was inhibited by phenylmethylsulfonyl fluoride, suggesting the enzyme to be a serine protease. The optimum temperature for the enzyme activity was 85 degrees C, and about 50% of the original activity remained after incubation at 90 degrees C for 10 min in the presence of Ca2+. The optimum pH for the enzyme activity was 11.0 and the enzyme was fairly stable from pH 5.0 to 12.0. The gene for this thermostable alkaline protease was cloned in Escherichia coli and the expressed intracellular enzyme was activated by heat treatment. Sequence analysis showed an open reading frame of 1,152 base pairs, coding for a polypeptide- of 384 amino acids. The polypeptide was composed of a signal sequence (25 amino acids), a prosequence (81 amino acids), and a mature protein of 278 amino acids. The deduced amino acid sequence of the mature protease had high similarity with thermitase, a serine protease from Thermoactinomyces vulgaris, and the extent of sequence identity was 76%. PMID- 8704315 TI - Pericytes from microvessel fragment produce type IV collagen and multiple laminin isoforms. AB - In the microvascular system, pericytes are located at the abdominal side of capillary endothelial cells. To discover the role of pericytes in the microvascular system, we have analyzed the extracellular proteins secreted from pericytes isolated from microvessel fragments of rat epididymal fat pads and found that they synthesize substantial amounts of basement membrane components such as type IV collagen and laminins. Secretion of type IV collagen was markedly stimulated by ascorbic acid phosphate. Reducing and nonreducing sodium dodecyl sulfate gel electrophoresis showed that pericytes produce six laminin chains assembled into different trimeric isoforms. Two of them were similar to laminin variants produced by aortic and pulmonal endothelial cells but others were suggested to be novel variants. PMID- 8704316 TI - Purification, characterization, and crystallization of single molecular species of beta-conglycinin from soybean seeds. AB - Four major molecular species of beta-conglycinin, alpha 3, alpha 2 beta, alpha beta 2, and beta 3, were isolated and purified from seeds of an alpha' subunit deficient strain of soybeans (Glycine max). All components were found to be homogeneous by high pressure liquid chromatography, SDS-polyacrylamide gel electrophoresis, and amino acid and amino terminal sequence analyses. The amino acid compositions of the alpha 3 and beta 3 components agreed fairly well with the compositions deduced from the cDNA sequences, and all of the components were highly glycosylated. The alpha 3 and beta 3 components were compared regarding their secondary structures. The secondary structure of the alpha 3 component deduced from CD measurements showed a higher alpha-helix content than that of the beta 3 component. The beta 3 component was crystallized by decreasing the ionic strength from 0.5 to 0.14 in phosphate buffer, pH 7.3, and the crystals grew to a size (1.0 mm x 0.2 mm x 0.2 mm) suitable for X-ray crystallographic analysis. A preliminary X-ray analysis showed that the crystal belonged to an orthorhombic crystal system having the space group P2(1)2(1)2(1) and unit cell dimensions of a = 185.1 A, b = 107.9 A, and c = 97.6 A. PMID- 8704317 TI - In vivo inhibition of kynurenine aminotransferase activity by isonicotinic acid hydrazide in rats. AB - It is known that the anti-tuberculosis drug, isonicotinic acid hydrazide (INH), causes pellagra, a niacin deficiency syndrome, and peripheral neuritis in humans. We investigated the effects of INH on the metabolism of tryptophan to niacin in rats fed on a niacin-free diet. The activity of kynurenine aminotransferase was significantly inhibited by feeding a diet containing INH and by an injection of INH, and the urinary excretion of xanthurenic acid, the side-reaction product of the conversion pathway of tryptophan to niacin, was below the limit of detection. The inhibition of kynurenine aminotransferase and the resulting decreased formation of xanthurenic acid generally mean a higher conversion ratio of tryptophan to niacin. However, the conversion ratio was no different between the control and INH groups. PMID- 8704318 TI - Immune functions of immunoglobulin Y isolated from egg yolk of hens immunized with various infectious bacteria. AB - We studied the immune functions of IgY obtained from hens immunized with a mixture of formalin-treated pathogenic bacteria. The IgY inhibited the growth of Pseudomonas aeruginosa, the production of Staphylococcus aureus enterotoxin-A, and adhesion of Salmonella enteritidis to cultured human intestinal cells (Caco 2). The results indicated that IgY specific for plural bacteria has effects useful toward prevention of bacterial diseases. PMID- 8704319 TI - Active transport activities of free B-6 vitamers in various yeast strains. AB - Active transport activities of free B-6 vitamers in 35 strains of 8 genera of yeast were measured by isocratic reverse-phase HPLC. Many but not all strains transported pyridoxamine and/or pyridoxine. The active transport activities in some yeast strains tested were completely inhibited by amiloride (0.5 mM). In contrast to cells so far studied, yeast cells showed a novel character in metabolism of accumulated B-6 vitamers: the phosphorylation of the free B-6 vitamers was regulated at a low level. There was no apparent correlation between the presence of the active transport activity in yeasts and the requirement of vitamin B-6 for their growth. PMID- 8704321 TI - Isolation and identification of two new diterpene glycosides from Nicotiana tabacum. AB - Two new diterpene glycosides containing 20-hydroxygeranyllinalool were isolated and identified from Nicotiana tabacum. These compounds consisted of five molecules of glucose and/or rhamnose. The locations of the aglycone and glycosides in the molecules were determined by 2D-NMR with the HMBC technique. The structures were (6E,10E,14Z)-20-hydroxygeranyllinalyl-3-O- [alpha-L rhamnopyranosyl (1 --> 4)]-beta-D-glucopyranoside-20-O- [beta-D-glucopyranosyl (1 --> 2)]-[alpha-L-rhamnopyranosyl (1 --> 6)]-beta-D- glucopyranoside and (6E,10E,14Z)-20-hydroxygeranyllinalyl-3-O- [alpha-L-rhamnopyranosyl (1 --> 4)] beta-D-glycopyranoside-20-O- [alpha-L-rhamnopyranosyl (1 --> 4)]-[alpha-L rhamnopyranosyl (1 --> 6)]-beta-D- glucopyranoside. PMID- 8704320 TI - Bestatin analogue from Streptomyces neyagawaensis SL-387. AB - A bestatin analogue, (2S,3R)-3-amino-2-hydroxy-4-phenylbutanoyl-L-valine (AHPA Val), from the culture filtrate of Streptomyces neyagawaensis SL-387 was obtained in a chemically defined medium containing DL-3-amino-3-phenylpropionic acid. AHPA Val was 6 times (IC50 = 1.2 micrograms/ml) as strong as bestatin (IC50 = 7.0 micrograms/ml) against porcine kidney microsomal aminopeptidase N, and 4 times (5.6 micrograms/ml) as strong as bestatin (IC50 = 20.7 micrograms/ml) against aminopeptidase N of human metastatic fibrosarcoma HT1080. To the best of our knowledge, this is the first report on the microbial production of AHPA-Val. PMID- 8704322 TI - Aclacinomycin X, a novel anthracycline antibiotic produced by Streptomyces galilaeus ATCC 31133. AB - A new anthracycline antibiotic, designated as aclacinomycin X, was isolated from the culture broth of Streptomyces galilaeus ATCC 31133, and was identified as 7 (O-rhodosaminyl-deoxyfucosyl-rednosyl)- aklavinone. Its in vitro cytotoxicity was tested against several human tumor cell lines. PMID- 8704324 TI - Rapid and sensitive screening of antifungal activity in medicinal plants by a single-cell biosensing system. AB - A biosensing system based on the response of fungal cells was used for the evaluation of antifungal activity of medicinal plants against Aspergillus niger. This system measured the hyphal growth rate in real time in the presence or absence of Chinese herbal extracts. The sensitivity of this system was 100-fold higher than that of conventional methods, and is advisable for the screening of antifungal compounds. PMID- 8704323 TI - Synthesis and antifungal activity of cinnamic acid esters. AB - Cinnamic, p-coumaric and ferulic acids were isolated from pineapple stems (Ananas comosus var. Cayenne). Twenty-four kinds of esters were prepared from these acids, alcohols and the components of Alpinia. Isopropyl 4-hydroxycinnamate (11) and butyl 4-hydroxycinnamate (12) were found to have almost the same effectiveness in antifungal activity against Pythium sp. at 10 ppm as that of the commercial fungicide iprobenfos (kitazin P). PMID- 8704326 TI - Geranyl 6-O-alpha-L-arabinopyranosyl-beta-D-glucopyranoside isolated as an aroma precursor from leaves of a green tea cultivar. AB - A new glycosidic aroma precursor was isolated from green tea leaves (Camellia sinensis var. sinensis cv. Yabukita) along with the known primeverosides of cis linalool 3,6-oxide, linalool and geraniol. These glycosides were separated by chromatographic isolation on Amberlite XAD-2, ODS flash chromatography, and finally HPLC. The chemical structure of the new unknown glycoside was confirmed as geranyl 6-O-alpha-L-arabinopyranosyl-beta-D-glucopyranoside (geranyl beta vicianoside) by spectrometric analyses and by an enzymatic hydrolysis with glycosidase followed by GC-MS and HPLC analyses. Moreover the vicianoside was hydrolyzed with acetone powder obtained from fresh tea leaves to generate the same compounds, suggesting this glycoside to be a tea aroma precursor. PMID- 8704325 TI - Construction and characterization of a deletion mutant of gpd2 that encodes an isozyme of NADH-dependent glycerol-3-phosphate dehydrogenase in fission yeast. AB - Schizosaccharomyces pombe has two genes each encoding an isozyme of NADH dependent glycerol-3-phosphate dehydrogenases (gpd1+ and gpd2+). To gain an insight into the function of these genes, here we constructed a gpd2 deletion mutant, in addition to the previously constructed gpd1 deletion mutant. We showed that the gpd1+ and gpd2+ gene-products are both functional in terms of the de novo glycerol synthesis. Furthermore, the gpd1(+)-mediated glycerol production is primarily responsible for the osmoregulation, but the gpd2+ gene is not. Interestingly, however, the gpd2 deletion mutant had histidine- or lysine auxotrophy for growth on a minimal medium. PMID- 8704327 TI - Digalactosyldiacylglycerol suppression of inhibition by sulfoquinovosyldiacylglycerol of alpha-glucosidase. AB - Digalactosyldiacylglycerol (DGDG) suppressed the inhibition by sulfoquinovosyldiacylglycerol (SQDG) of an alpha-glucosidase reaction. Suppressing was considered to be an apparent decrease in inhibitory activity of SQDG ascribed to direct interaction between SQDG and DGDG. This suppression was presumed to be caused by less access of SQDG to the enzyme because SQDG and DGDG formed mixed micelles. PMID- 8704328 TI - Roles for theatre nurses--feedback. PMID- 8704329 TI - To sterilise or disinfect--that is the question. AB - Few hospitals have the in-house technology to sterilise heat and moisture sensitive reusable devices, yet the use of this type of equipment is increasing in all types of surgery. In general terms and because of limitations of existing technology, instruments and equipment have been divided into three categories based on the degree of risk of infection involved in their use: 1. Instruments and equipment which come into contact with intact skin must be cleaned before they are used. 2. Instruments and equipment which come into contact with non sterile tissue (other than intact skin) must be disinfected before they are used. 3. Instruments and equipment which enter, or are capable of entering, the vascular system or tissue that would be sterile under normal circumstances must be sterilised before they are used. Unfortunately, some devices that should be sterilised are only disinfected in liquid chemical disinfectants. Traditional low temperature sterilisation systems such as ethylene oxide (EtO) and low temperature steam and formaldehyde (LTSF) have many disadvantages and it is undoubtedly for this reason they have been relegated to a few specialist sterile service departments in the UK. PMID- 8704330 TI - Reflections--glutaraldehyde glooms. PMID- 8704331 TI - Hypnotherapy and acute pain control. AB - With the increasing acceptance of various forms of alternative medicine as coping strategies for the stresses and strains of modern life, hypnotherapy is becoming more and more established as an adjunct to orthodox medical practice. Scientific research methods applied to hypnotism, as well as improved communication and education, have done much to raise the image of hypnotherapy generally. PMID- 8704332 TI - Modern apprenticeships in the NHS. Operating department practice. PMID- 8704333 TI - Pain relief after thoracotomy. PMID- 8704334 TI - Helicobacter pylori--the hidden infection. AB - Surgical intervention is the failure of medicine. Or so it was once said. Those who have been involved in the treatment of peptic ulceration would probably agree with that statement. In the Fifties, general surgery lists included a high proportion of gastric surgery. Between then and now there has been an increasing refinement of techniques until the development of more effective medical treatment led to a reduction in surgery for peptic ulceration. In recent years the realisation that the micro-organism helicobacter pylori is significant in the incidence of ulcers has opened up new lines of treatment. Understanding of this infection is still necessary for those involved in therapeutic and diagnostic surgical interventions. PMID- 8704335 TI - Theatre nursing in Germany. AB - I visited the operating department at the Paracelcus Klinic in Osnabruck, Northern Germany as part of a visit organised within the Erasmus scheme funded by the European Union. Although now a nurse teacher, I was formerly a theatre sister and continue to liaise with the area, and so I was pleased to have the opportunity to see how an operating department in Germany was organised. PMID- 8704336 TI - Some dilemmas of living wills. AB - The question of care of terminally ill patients is, as we all know, fraught with difficulties which become more rather than less intense with new developments in medical systems. Once, doctors were not faced with the question of whether to keep someone alive when it was scarcely in their power to intervene in nature's course. Today, with new powers to keep people alive, the question arises, with ever greater urgency, when and whether those powers should be used. PMID- 8704337 TI - On the other side. PMID- 8704338 TI - Back in the fold! PMID- 8704339 TI - Glove powder and latex allergy. PMID- 8704340 TI - Red rubber double-lumen endobronchial tubes. PMID- 8704341 TI - Essentials of anaesthesia for opthalmic surgery. AB - Unfortunately the eye is prone to very many pathological conditions, either congenital or acquired. Thankfully, all but the most extreme are amenable to surgical treatment. The anaesthetist's role in this process is, principally, to present the surgeon with an eye which is operable. How this is done will be considered in the latter part of this article. However, as in all areas of anaesthetic practice, a detailed knowledge of anatomy and physiology is essential. This aspect will be dealt with first. PMID- 8704342 TI - Peri-operative care of the patient with diabetes mellitus. AB - There are a number of factors to take into consideration when a patient with diabetes has to have an operation. Care of the patient with diabetes is tailored to individual needs and this care needs to extend right through their time in hospital, including pre-operative, peri-operative and post-operative care. This paper examines the rationale for care of the diabetic patient in the operating department and will suggest a protocol for through theatre care. PMID- 8704343 TI - Managing hypovolaemic shock. AB - This poster has been designed to impart information which was gathered for a seminar as part of an Anaesthetic Nursing Course. It looks at the research which has been compiled on this subject, both in the Nursing and the Medical fields and illustrates the recommendations made by these pieces of research. The poster also depicts the breakdown of fluid compartments in the body and the physiological transference of fluids within the body in times of crisis. There is another section in the poster which explains the differences between certain fluids which are typically used during hypovolaemic resuscitation and the reasons for using these particular fluids. A version of a table designed by the American College of Surgeons, (Advanced Trauma Life Support, ATLS), which is recommended in some of the research on this topic is also shown in the poster and is very useful as a guide to what should happen in the hypovolaemic situation. PMID- 8704344 TI - Haemostasis. AB - There are many different methods and techniques used to achieve this in operating theatres, day procedure units and general practice surgeries around the country. I am aware that some of the methods which I am going to mention may not be available in all work areas but they are techniques which are synonymous with achieving haemostasis and are methods which you may want implemented in your area. So let me outline the topics I intend to look at in this session. First of all I would like to briefly outline the basic physiology behind coagulation, the body's natural response to bleeding. Then, I would like to mention a few drugs which can affect haemostasis and also look at the use of electrosurgery in achieving haemostasis. I would then intend to look at a few other techniques which are used such as suturing, tourniquets and pressure dressings. PMID- 8704345 TI - Homeostasis--the key concept to physiological control. PMID- 8704346 TI - Animal to human transplants: the ethics of xenotransplantation (1). AB - There now exists, potentially, the scientific and medical knowledge to transplant animal organs into humans, a procedure called xenotransplantation. The rationale behind the idea is that demand for transplanted human organs far exceeds supply. For this reason, alternative methods have been sought. The use of animals is seen as one way to deal with the problem. PMID- 8704347 TI - Infection control and universal precautions. AB - Surgical interventions are now carried out in many settings other than purposely designed operating theatre suites. NATN is pleased to welcome nurses working in areas other than operating theatres to associate membership of the Association. In response to this diversity of surgical settings, an examination of the basic principles of infection control and the application of universal precautions may be valuable. PMID- 8704348 TI - Positive interaction between 17 beta-Estradiol and parathyroid hormone in normal human osteoblasts cultured long term in the presence of dexamethasone. AB - We previously developed two models of human osteoblasts with distinct differentiation stages using cells derived from iliac crest trabecular bone explants cultured long term in the presence (HOB + DEX) and absence (HOB - DEX) of 10 nM dexamethasone (DEX) (Wong et al., J Bone Miner Res 1990;5:803). Using these models from 36 subjects aged 41-80 years, we examined the effects of 17 beta-estradiol (E2) on cell proliferation, osteocalcin (OC) production, alkaline phosphatase (ALP) and basal and parathyroid hormone (PTH)-stimulated adenylate cyclase activities, as well as the steady-state mRNA levels of ALP, collagen type I(COLL), OC, and receptors for E2 (ER) and PTH (PTHr). E2 alone had no effect on [3H]thymidine uptake in (HOB - DEX) cells but appeared to stimulate the uptake in (HOB + DEX) cells in a dose-dependent manner, with maximum effect at 10(-10)M (p < 0.05). However, in the presence of 10(-6)M PTH, E2 inhibited the uptake in (HOB - DEX) cells (ANOVA, KW = 18.95, p < 0.005) but stimulated the uptake in (HOB + DEX) cells (KW = 13.52, p < 0.025). E2 decreased the amount of osteocalcin in culture media from both (HOB - DEX) and (HOB + DEX) cells (p < 0.05). PTH alone or E2, alone or in combination with 10(-9)M PTH, had no effect on ALP activity in (HOB - DEX) cells. In contrast, in (HOB + DEX) cells, E2 + PTH but not E2 alone, had biphasic effects on ALP activity, with maximum stimulation observed at 10( 11) and 10(-10)M E2, and a return to basal levels at 10(-9)M E2. E2 decreased basal adenylate cyclase activities in a dose-dependent manner in (HOB + DEX) but not (HOB - DEX) cells (KW = 13.48, p < 0.05). In (HOB + DEX) cells, E2 had biphasic effects on PTH-stimulated adenylate cyclase activity, with significant stimulation observed at 10(-10)M (p < 0.05). While E2 had no significant effect on osteoblastic marker mRNA levels in (HOB - DEX) cells, it decreased osteocalcin and stimulated PTHr mRNA levels in (HOB + DEX) cells. Thus, in our human osteoblastic cell models, estrogen regulated metabolic function largely in the more differentiated cells, by modifying the effects of PTH. PMID- 8704349 TI - Screening for osteopenia and osteoporosis: selection by body composition. AB - There is a great need for simple means of identifying persons at low risk of developing osteoporosis, in order to exclude them from screening with bone mineral measurements, since this procedure is too expensive and time-consuming for general use in the unselected population. We have determined the relationships between body measure (weight, height, body mass index, lean tissue mass, fat mass, waist-to-hip ratio) and bone mineral density (BMD) in 175 women of ages 28-74 years in a cross-sectional study in a county in central Sweden. Dual-energy X-ray absorptiometry was performed at three sites: total body, L2-4 region of lumbar spine, and neck region of the proximal femur. Using multiple linear regression models, the relationship between the dependent variable, BMD, and each of the body measures was determined, with adjustment for confounding factors. Weight alone, in a multivariate model, explained 28%, 21% and 15% of the variance in BMD of total body, at the lumbar spine and at the femoral neck according to these models. The WHO definition of osteopenia was used to dichotomize BMD, which made it possible, in multivariate logistic regression models, to estimate the risk of osteopenia with different body measures categorized into tertiles. Weight of over 71 kg was associated with a very low risk of being osteopenic compared with women weighing less than 64 kg, with odds ratios (OR) of 0.01 (95% confidence interval (CI) 0.00-0.09), 0.06 (CI 0.02-0.22) and 0.13 (CI 0.04-0.42) for osteopenia of total body, lumbar spine and femoral neck, respectively. Furthermore a sensitivity/specificity analysis revealed that, in this population, a woman weighing over 70 kg is not likely to have osteoporosis. Test specifics of a weight under 70 kg for osteoporosis (BMD less than 2.5 SD compared with normal young women) of femoral neck among the postmenopausal women showed a sensitivity of 0.94, a specificity of 0.36, positive predictive value (PPV) of 0.21, and negative predictive value (NPV) of 0.97. Thus, exclusion of the 33% of women with the highest weight meant only that 3% of osteoporotic cases were missed. The corresponding figures for lumbar spine were sensitivity 0.89, specificity 0.38, PPV 0.33, and NPV 0.91. All women who were defined as being osteoporotic of total body weighed under 62 kg. When the intention was to identify those with osteopenia of total body among the postmenopausal women we attained a sensitivity of 0.92 and a NPV of 0.91 for a weight under 70 kg, whereas we found that weight could not be used as an exclusion criterion for osteopenia of femoral neck and lumbar spine. Our data thus indicate that weight could be used to exclude women from a screening program for postmenopausal osteoporosis. PMID- 8704350 TI - Effect of ovariectomy on intraosseous vascularization and bone remodelling in rats: action of tiludronate. AB - In order to study the action of tiludronate on the changes in intraosseous vascularization induced by ovariectomy, and to link these effects to those observed in bone remodelling, 30 female Sprague-Dawley rats (age 40 weeks) were studied. Ten rats were shamoperated and treated by vehicle, 10 rats were ovariectomized and treated by vehicle, and 10 rats were ovariectomized and treated orally with tiludronate, 0.16 mmol/kg/per day, 3 days a week for 16 weeks, from the day following ovariectomy. The rats were killed after 4 months, and a histomorphometric study and quantification of intraosseous vessels carried out on the sixth lumbar vertebra. The area of the intraosseous sinusoidal capillaries increased after ovariectomy, which also induced a moderate increase in resorption surfaces and osteoid surfaces leading to a decrease of 40% in the trabecular bone volume at the lumbar spine level. This bone mineral loss was completely prevented by tiludronate, which normalized the bone turnover. However, tiludronate was without any effect on intraosseous vascularization. These results indicate that the surface area of the intraosseous sinusoidal capillaries was correlated positively with resorption surfaces and negatively with trabecular bone volume and the number of bone trabeculae. In these experimental conditions, an inhibitor of bone resorption can exert its positive effect on bone mass without normalization of vascularization. PMID- 8704352 TI - Geometric measurements of the proximal femur in UK women: secular increase between the late 1950s and early 1990s. AB - The aim of this study was to determine whether hip axis or femoral length has increased in women in the United Kingdom between the late 1950s and early 1990s. Such an observation would be of interest as it might explain the rise in age specific incidence of hip fracture observed during these years. We studied two sets of antero-posterior pelvic radiographs of women aged 55-69 years taken during the course of population-based studies in the UK, one in 1958-60 and the other in 1989-91. One observer (S.G.) recorded the following measurements at the right hip: hip axis length (HAL), femoral length (FL) and femoral width (FW). Two summary ratios, HAL/FW and FL/FW were calculated to allow for differences in radiographic technique. HAL, FL and FW were greater in the 1989-91 films compared with those taken in 1958-60. Both HAL and FL expressed as a ratio to FW were also greater in the later films. FL/FW increased by 4.5% (p < 0.05); HAL/FW increased by 2.3%, though this was not statistically significant. We conclude that there has been a small apparent change in geometric measurements of the hip during the past 36 years. Cautious extrapolation suggests that such a change may explain up to one third of the increase in incidence of hip fracture observed during this period. PMID- 8704351 TI - The association of patellar ultrasound transmissions and forearm densitometry with vertebral fracture, number and severity: the Saunders County Bone Quality Study. AB - Ultrasonic measures of bone have been available for clinical research purposes for nearly 10 years, yet there still seems to be a need to compare ultrasound with the accepted gold standard of densitometry. Recently there have been published reports showing that ultrasound measures are associated with both appendicular and hip fracture, in particular after adjustment for densitometry measures. We present here a comparison between speed of sound through the patella and forearm bone densitometry, using their association with prevalent vertebral fractures in a population-based study of women and men. The prospective phase of the Saunders Bone Quality Study includes 1401 women and men who had baseline spine radiographs, patellar ultrasound, and forearm densitometry measurements. Multivariate forward logistic regression was used to determine the age-adjusted odds of vertebral fracture, the number of fractures, and the severity of these fractures, when patellar ultrasound and each of four forearm densitometry measures were entered into the model. Age is the most important factor associated with vertebral fractures, their number, and severity for women, while age is not significantly related to vertebral fractures for men. Of the bone status measures, patellar ultrasound entered the logistic regression models more consistently than any other measure except ulnar bone mineral density for women. The ultrasound measure entered every model for men. We conclude that patellar ultrasound velocity is more consistently associated with the odds of vertebral fractures than radius bone mineral content. PMID- 8704353 TI - Bone mineral acquisition during adolescence and early adulthood: a study in 574 healthy females 10-24 years of age. AB - Low bone mass is known to be associated with an increased risk of fractures. Osteoporosis prevention by maximizing bone mass will be crucial and requires a better knowledge of bone mass acquisition during adolescence. Bone mass was assessed in 574 healthy volunteer females aged 10-24 years. Spine bone mineral density (BMD) in anteroposterior (AP L2-4) and lateral (LAT L3) views was measured using dual-energy X-ray absorptiometry (DXA) and AP bone mineral content (BMC) was calculated. At the same time, spine AP-BMD (L2-4) was evaluated in 333 normal menstruating women, aged 27-47 years. Bone values, osteocalcin and IGF-1 serum concentrations were correlated with chronological age, skeletal age, pubertal stages and time after menarche. In this cross-sectional study, AP- and LAT-BMD and BMC increased dramatically between skeletal ages 10 and 14 or until the first year after menarche. Between 14 and 17 skeletal years of age, AP-BMD and BMC increased moderately, whereas LAT-BMD remained unchanged. After skeletal age 17, or the fourth year after menarche, there was no significant increase in BMD or BMC, and their values did not differ from those of menstruating women. A serum osteocalcin peak was observed at skeletal ages 11-12 or at stage P3, whereas IGF-1 peaked at 13-14 skeletal years of age or at P4 and the first year after menarche. Eighty-six per cent of the adult bone mass of the spine is acquired before skeletal age 14 or the second year after menarche; therefore osteoporosis prevention programs will be particularly effective before that age. PMID- 8704354 TI - Caffeine does not affect the rate of gain in spine bone in young women. AB - The effect of nutrition and dietary caffeine consumption and physical activity on bone gain in women during the third decade of life was determined in a longitudinal, descriptive study of 145 healthy college-aged women. Estimates of caffeine and other dietary intakes were determined by repeated 7-day diet diaries. Measurements of bone mineral in the spine and total body mineral content were determined by dual-photon absorptiometry. Measures of physical activity were ascertained by physical activity monitor. The mean estimated caffeine, calcium and protein intakes for the young women students were 103 +/- 106 mg/day (mean +/ SEM), 831 +/- 334 mg/day (mean +/- SEM) and 66 +/- 16 g/day (mean +/- SEM) respectively. The median rates of bone gain were 5.9% for spine bone mineral content, 6.8% for spine bone mineral density and 12.5% for total body bone mineral. In a multiple regression analysis the significant predictors (+ or -) of the rate of gain were age (-), activity (+), calcium intake (+) and protein intake (-). Caffeine consumption was not associated with significant reduction in rates of bone gain. While calcium and protein nutrition affect bone gain in the third decade of life in women, moderate caffeine intake (one cup of coffee per day, or 103 mg) appears to be safe with respect to bone health in this age group. PMID- 8704355 TI - Bone turnover and density in healthy women during breastfeeding and after weaning. AB - To investigate the changes in maternal bone density and turnover associated with lactation we ran a longitudinal study in fully breastfeeding women (age 26.3 +/- 4.1 years, mean +/- SD) at the first (stage I, n = 30) and sixth (stage II, n = 25) months postpartum and 6 months after weaning (stage III, n = 20), and in a contemporary control group of non-nursing women. At each time point bone density, serum calcium, phosphorus, alkaline phosphatases, parathyroid hormone (PTH), osteocalcin, follicle stimulating hormone (FSH), estradiol (E2), prolactin (PRL) urinary hydroxyproline and creatinine (OH-P/Cr) were measured in both groups. The daily calcium intake of nursing women (1479 +/- 590 mg/day at stage I) was higher than in non-nursing women (536 +/- 231 mg/day at stage I). Biochemical markers of bone turnover were higher (p < 0.05) in nursing than in non-nursing women at stages I and II, while in stage III only OH-P/Cr was elevated. The lumbar spine (L2-4) bone mineral density was similar in the two groups at the beginning of the study (1.148 +/- 0.111 g/cm2 in nursing women vs 1.211 +/- 0.102 g/cm2 in non nursing women; p = 0.06), but it was lower in nursing women at stage II (1.144 +/ 0.110 g/cm2 vs 1.216 +/- 0.095 g/cm2 respectively; p < 0.05). Right femoral neck bone density decreased by 3% between stages I and II in nursing women but did not differ from values in non-nursing women (0.947 +/- 0.110 vs 0.973 +/- 0.108 in stage I and 0.918 +/- 0.114 vs 0.975 +/- 0.098 in stage II respectively; p < 0.05, ANOVA). After weaning, lumbar spine and femoral neck bone density increased by 6% and 8% respectively (p < 0.05, ANOVA). No correlation was found between changes in bone turnover markers or bone density and parity, frequency and duration of nursing episodes, body weight, body mass index, and plasma PRL, E2 and PTH levels. We conclude that in nursing women with a daily calcium intake at the recommended dietary allowance ( > 1200 mg/day), full breastfeeding extending over 6 months is characterized by increased maternal bone turnover and a transient bone loss which normalizes after weaning. PMID- 8704356 TI - Prevalence and severity of vertebral fracture: the Saunders County Bone Quality Study. AB - Vertebral fracture prevalence and severity were analyzed by sex and age in an age stratified proportionate sample of the enumerated population of women and men 50 years of age and older in Saunders County, Nebraska. The sample consisted of 899 women and 529 men. Of these, all but 10 women and 2 men had readable lateral spine radiographs. For both sexes, fracture prevalence rises with age. Women in their fifties have 10% vertebral fracture prevalence, and women in their eighties, 45% prevalence. Men in their fifties have 29% prevalence, and men in their eighties, 39% prevalence. The rise in prevalence and total spinal deformity with age is much greater for women than for men, but the prevalence of vertebral deformity in the fifties is much greater in men than in women. PMID- 8704357 TI - The bisphosphonate ibandronate, given daily as well as discontinuously, decreases bone resorption and increases calcium retention as assessed by 45Ca kinetics in the intact rat. AB - The new bisphosphonate ibandronate was given at various doses and regimens to normal growing rats, and its effect on calcium metabolism investigated by means of 45Ca kinetics. The bisphosphonate began to inhibit bone resorption at a dose of 0.1 microgram P/kg, given daily. At higher doses intestinal calcium absorption, calciuria and calcium balance were also increased, calcemia being decreased. There was no difference in effect when the same amount of compound was given either daily for 10 days or all at once. Furthermore, the effect of a high dose of 100 micrograms P/kg was present 1 month after a single administration, whereas a dose 10 times lower was no longer effective. These results suggest that ibandronate may be effective in humans for decreasing bone resorption and increasing calcium balance in osteoporosis, when given either daily or discontinuously. PMID- 8704359 TI - The genetics of proximal femur geometry, distribution of bone mass and bone mineral density. AB - To estimate genetic effects on femoral neck geometry and the distribution of bone mineral within the proximal femur a cross-sectional twin analysis was carried out at a university hospital that compared correlations in these traits in pairs of mono- and dizygotic female twins. Monozygotic (MZ, n = 51 pairs, age 49.1 +/- 9.3 years) and dizygotic (DZ, n = 26 pairs, age 45.7 +/- 11.3 years) twins were randomly selected from a larger sample of twins previously studied. Measurements of bone mineral density (BMD), femoral neck angles and length, cross-sectional area and moment of interia, the center of mass of the narrowest cross-section of the femoral neck, and BMDs of regions within the femoral neck were made. A summary index of the resistance of the femoral neck to forces experienced in a fall with impact on the greater trochanter (Fall Index, FI) was calculated. MZ pair intraclass correlations (rMZ) were significantly (p < 0.05) different from zero for all bone mass and femoral geometry variables (0.35 < rMZ < 0.82). DZ pair correlations (rDZ) were lower than rMZ for all variables (0.04 < rDZ < 0.52) except femoral neck length (rDZ = 0.38, rMZ = 0.36). After adjustment for BMD of the femoral neck, rMZ was significantly greater than rDZ, yielding high heritability estimates for regional BMDs (0.72 < H2 < 0.78), the center of mass of the femoral neck (H2 = 0.70, -0.04 to 1.43 95% CI) and the resistance of the femoral neck to forces experienced in a fall (FI, H2 = 0.94, 0.06 to 1.85 95% CI), but not for femoral neck length. Adjustments for age did not alter these findings. It is concluded that there are significant familial influences on the distribution of femoral bone mass and on the calculated structural strength of the proximal femur, but not on femoral neck length. If the assumptions of the twin model are correct, this is evidence for genetic factors influencing these traits. PMID- 8704358 TI - In corticosteroid-treated respiratory diseases, monofluorophosphate increases lumbar bone density: a double-masked randomized study. AB - The efficacy of a monofluorophosphate-calcium combination (MFP-Ca) in increasing lumbar bone mineral density (BMD) was assessed in a prospective double-masked study. Patients (n = 35), who had been treated for 1 year or more with prednisone equivalent doses > or = 7 mg/day for asthma or other respiratory diseases, were randomly assigned to receive twice a day, for 2 years, either one MFP-Ca tablet [100 mg sodium monofluorophosphate (13.2 mg F-) + 500.5 mg Ca2+] or one Ca tablet (500.5 mg Ca2+). BMD was measured from L2 to L4 using a dual photon absorptiometer. The eligible patients (7 premenopausal women, 21 men), who had no previous vertebral fractures and were aged 46.5 (21-65) years, had received 18 (7.5-60) mg prednisone-equivalent/day and had a mean lumbar BMD of 0.917 +/- 0.141 g/cm2 at baseline (MO); in these 28 patients, the mean increase in lumbar BMD at final assessment was significantly greater in the MFP-Ca group (p = 0.05; Mann-Whitney). There was also a significant difference after 2 years between the two groups (p = 0.05, ANOVA) in favour of MFP-Ca, with an increase in lumbar BMD of 11% (MFP-Ca) compared with 1% (Ca); thus, with MFP-Ca, lumbar BMD increased by an average of approximately 5.5%/year. There was no statistically significant difference between the two groups in doses of corticosteroids used during the 2 study years, rate of vertebral fractures, or frequency of side-effects (which were all minor). No bone fissure was observed. Thus, the daily dose of 200 mg monofluorophosphate (26.4 mg F-) combined with 1 g Ca2+ in patients with long term corticosteroid-treated respiratory diseases appears to be a safe and efficient way of increasing lumbar BMD, suggesting that its use should be further studied in corticosteroid-induced osteoporosis. PMID- 8704360 TI - The European spine phantom: reply to Dr Peter Tothill's letter. PMID- 8704361 TI - Skin thickness measured by ultrasonography (US) and bone mineral density (BMD) in 94 healthy postmenopausal women. PMID- 8704362 TI - [Strategies for productive collaboration between science and general practice]. AB - My contribution is a systematic account of strategies for cooperation and feedback between public health sciences and research. A feature both areas have in common is their commitment to empirically based knowledge. "Rational" models of cooperation (or rather, models based on belief in rationality) presume simple linear links between research and its application. Diffusion models, however, seem to be far more adequate for translating public health research into reality. The best model is the "Discourse Model" based on continuous communication and feedback between science and practice. It seems to promote the diffusion of public health innovations most effectively. In the second part I give examples of how to translate the Discourse Model into practical reality. Summing up, its strategic elements are: 1. responsive project shaping, 2. interactive running of projects, 3. shared processing of results, 4. active diffusion of results. PMID- 8704363 TI - [Family practice management of risk patients with reference to preclinical emergency medicine]. AB - Chances of surviving myocardial infarction or cardiocirculatory arrest depend on an early and appropriate emergency call as well as on bystander CPR. This study uses a questionnaire to assess to what extent these factors guide the private practitioners caring for high-risk cardiac patients. Almost all physicians responding (44%) treat such patients. More than 70% try to include the patients' relatives in a medical briefing. Recognition of early warning signs is discussed with 80% of the patients and 70% of relatives. How to identify acute life threatening conditions is discussed with 60% of patients and relatives. In case of sudden chest pain and dyspnoea, physicians commonly advise calling the family doctor, a hospital or a regional physician on call. Calling 112, the emergency physician/rescue service, is rarely recommended. Initiating CPR is recommended for an emergency by 30% of the physicians, whereas 20% encourage relatives of patients at risk to take a CPR course. CPR instruction is judged to be generally useful by 50% of physicians, of limited use by 40% and of no use by 10%. When asked why they do not encourage CPR, 15% of physicians said they have no time, 25% cited the inefficiency of the courses, 40% said CPR is psychologically too stressful for laypersons, and 50% felt it was physically too stressful. This study points out the necessity of additional, appropriate instruction of laymen regarding cardiac emergencies. PMID- 8704364 TI - [Empirical analysis of expert social medicine assessment of disability based on the decision of the Federal social court]. AB - Objective assessment of need for nursing care should be based on well defined criteria. In 1993, the Federal German Social Court ("Bundessozialgericht", BSG) has given criteria for identifying individuals in permanent need of very intensive nursing care ("Schwerpfledgebedurftige"), who qualified for benefits granted by the Germany statutory health insurance system at that time. The criteria were primarily based on the number of activities of daily living for which the applicants were in need of help. The criteria were largely, but not entirely, consistent with previously established criteria employed by the Medical Service of the Health Insurances ("Medizinischer Dienst der Krankenversicherung", MDK), who was responsible for the assessment. An analysis of 4185 examinations carried out by MDK in the region of Augsburg in 1991-1993 showed that the majority of judgments were in agreement with the criteria given by BSG. However, individuals with psychiatric disorders or symptoms were often rated by MDK as being in permanent need of very intensive nursing care even if BSG criteria were not fulfilled. Exclusive judgement of nursing dependency on the basis of single activities of daily living appears to be inadequate for this group of persons. PMID- 8704365 TI - [Why do elderly couples use ambulatory nursing care?]. AB - More than one million people in Germany are in need of home care and elderly caregiving wives and their caretaking husbands are an important group in the society. Demographical changes with a shift to more elderly people in relation to younger people will further increase the importance of this group. Home care service organisations play an important role in supporting these couples, but they are still rarely made use of. To optimize the support that can be rendered by home care services, information is needed on what influences the decision of elderly couples to use or not to use such services. A set of data from an earlier study about the situation of more than 300 couples aged 60 years or older is used to determine influencing factors in bivariate and multivariate analysis. The theoretical framework is a behavioural model of service use. The need for care measured by loss of function in activities of daily living turns out to be the most important factor influencing the use of home service organisations. Other variables such as e.g. disability of caregivers play a minor role. It seems that home care services are used as a kind of "last resort" if there is no other way to handle the situation. In trying to optimize home care services, the different values of factors influencing their use should be considered. It is important to take into consideration that those who solicit home care aid wish to retain their autonomy at the same time. PMID- 8704366 TI - [Criteria for admission of drug dependent criminals for withdrawal treatment according to section 64 StGB with reference to the newest legal guidelines]. AB - Due to a sentence of the German Federal Court of constitution from March 16th, 1994 requirements on the contents of expert opinions on drug or alcohol dependent delinquents considered for being sentenced to withdrawal treatment have changed. Up to that date it has to be stated that the prognosis of withdrawal does not seem to be useless at all. Later experiences with these dependent patients are often disappointing, mostly due to a lack of any therapeutical motivation. Requirements on prognostical criteria have now profoundly changed. The psychiatric expert must positively point out now that predictors of a positive outcome of withdrawal therapy really does exist. A marked decrease in frequency of these legal sentences to withdrawal therapy must be expected as a consequence of altered prognostical criteria. PMID- 8704367 TI - [Microbiological public health aspects in the use of rain water as water reservoirs for toilet flushing, garden irrigation and laundry]. AB - From a total of 102 rain water cisterns in use for toilet flushing, garden irrigation and laundering washing about 1,600 water samples were collected and subjected to microbiological analysis. The assays included aerobic heterotrophic microorganisms growing at 20 and 37 degrees C, respectively, as well as the identification of Escherichia coli, coliform organisms, faecal streptococci, Pseudomonas aeruginosa, staphylococci, yersiniae, salmonellae, shigellae, legionellae and yeasts. The median of the total number of cells per ml was 1,200 at 20 degrees C and 230 at 37 degrees C, respectively. Approximately 26 E. coli cells and 198 coliform organisms (median values) were found per 100 ml. In the case of cisterns manufactured of plastic the total number of cells was generally found to be lower than in samples collected from concrete or brick-made storage tanks. With the exception of the ubiquitously distributed organism Pseudomonas aeruginosa (found in 11.8% of the samples) and salmonella in only one sample, no other pathogens were detected. More than 95% of all analysed samples met the quality standards for bathing waters as set by the European Community. Provided certain precautions are taken, such as strict separation of mains for drinking water and rain water, as well as correct labelling of pipelines and collection sites, the use of rain water for toilet flushing, garden irrigation and laundry washing presents no unacceptable risk to public health. PMID- 8704369 TI - Certificate program in holistic nursing. Healing touch workshops. PMID- 8704368 TI - [Comprehensive and competition-oriented quality management in social medicine expert services]. AB - In free competition expert services in Social Medicine must supply their expertise with high quality in a short time and at low cost. The demands by customers in respect of motivation of the staff and innovative organisation are as important competitive factors as high quality standards for expertise production. These guiding principles completed by "Kaizen" and "Lean production" are necessary requirements for the further existence of the enterprise in competition. Quality assurance must be promoted in a process looking to the future in active quality management. PMID- 8704370 TI - Healing in love. PMID- 8704372 TI - Aromatherapy growing rapidly. PMID- 8704371 TI - Deeper states of consciousness--are they all the same? PMID- 8704373 TI - An unexpected love affair. PMID- 8704374 TI - Delegation of school health services to unlicensed assistive personnel. A position paper of the National Association of State School Nurse Consultants. PMID- 8704375 TI - Food choices of healthy school-age children. AB - This study examined the food choices of children 10 to 12 years of age to assess their knowledge of healthful food items. The subjects were 127 fifth and sixth grade boys and girls from public and private school systems. Data were gathered in the classroom. Students were asked to select which one of each of two dozen pairs of food items is better for their health. Results of an item analysis indicated that the majority of children were informed concerning healthful food choices, but they were unable to consistently select the healthier item. The findings suggest a need for additional educational programs for school-age children in the area of nutrition. PMID- 8704376 TI - School nurses' competence in caring for students who depend on medical technology. AB - This study explored the experiences of school nurses in giving care to students who depend on medical technology. From it, insight was gained into the nurses' competence in areas related to several aspects of care, ranging from planning to delegation. Six contributing factors to school nurses' competence were identified: challenge and excitement; education and experience; preparation and planning; collaboration with others; availability of expert resources; and reinforcement from successful outcomes. PMID- 8704378 TI - Funding resources for computers and peripherals. PMID- 8704377 TI - The child with HIV infection. PMID- 8704379 TI - Clonidine. PMID- 8704380 TI - Drug and alcohol use of school-age children in a rural community. AB - Drug abuse affects the physical and psychological well-being of the user as well as creating heavy societal costs--which are amplified if the user is a child. This study examined the relationship between drug and alcohol use and self esteem, school climate, peer effects, and family climate among a small sample of fourth graders in a rural community. PMID- 8704381 TI - Handwashing education can decrease illness absenteeism. AB - A handwashing program for elementary school students was developed and implemented by the school nurse in a suburban elementary school. The program consisted of surveying teachers, inspecting handwashing facilities, and providing classroom presentations and follow-up activities. Absenteeism records indicated a significant decrease in absenteeism for illness during the two months following the presentations. PMID- 8704382 TI - Delegation and supervision in school settings. Standards, issues, and guidelines for practice (Part 2). PMID- 8704383 TI - Asthma medications: a quick review. PMID- 8704384 TI - The significance of nursing classification systems to school nursing. PMID- 8704385 TI - Nursing practice management: muscular dystrophy. AB - The Nursing Practice Management section displays a health care plan for a student with Duchenne Muscular Dystrophy. Following a brief overview of the pathophysiology of the disorder, a case study is presented from which a nursing process-based plan of care is derived. PMID- 8704387 TI - Casemix update: Australian critical care costs and service weights. Part 1. PMID- 8704386 TI - The evolution of the school nurse practitioner: past, present, and future. AB - The role of school nurse practitioner (SNP) evolved in response to the growing need of the school-age population for health care and the school as a setting for providing primary health services. SNPs have broadened the role of the traditional school nurse while extending advanced nursing into the arena of primary health services. PMID- 8704388 TI - Preparation of the critically ill for transfer in metropolitan Sydney. AB - The transfer of the critically ill patient is often complex and difficult, and may contribute to increased morbidity. It is therefore valuable to have an understanding of the practical problems relating to patient transfer, and common management strategies, in order to minimise adverse outcomes. Key issues in the process of transporting the critically ill patient in metropolitan Sydney include: (i) particular problems and pitfalls during the transfer process; (ii) risk factors in transferring patients and the appropriateness of escorts and transfer procedures; (iii) procedures for arranging a patient transfer; (iv) the role of the NSW Medical Retrieval Co-ordination Centre (MRCC); (v) preparation of the patient for transfer, (vi) education of staff on the concept of safe matching of patient needs to appropriate escorts; and (vii) the need for a co-ordinated patient transfer system to Sydney metropolitan hospitals. An understanding of these issues, and an appropriate response to them from hospital staff will ensure safe and efficient transportation of critically ill patients in metropolitan Sydney. PMID- 8704389 TI - Chest X-ray quiz: Hampton's hump. PMID- 8704390 TI - Comparison of pulmonary artery pressure measurements in the supine and 60 degrees lateral positions. AB - Pulmonary artery pressure monitoring, with the patient in both the supine and lateral positions, is an essential element in the assessment of critically ill patients. Previous work offers conflicting results regarding the accuracy of measurements obtained with the patient in the lateral position. The purpose of this study was to determine if accurate pulmonary artery pressure measurements can be obtained in the cardiac surgical patient. Thirty-five patients underwent repositioning between the supine and both the left and right 60 degrees lateral position while being mechanically ventilated and then breathing spontaneously. Pulmonary artery pressure measurements were recorded prior to, two minutes following and ten minutes following repositioning. Despite some variation in results the pulmonary capillary wedge pressure measurement was reliable ten minutes after repositioning in both the spontaneously breathing and mechanically ventilated patient. Other pulmonary artery pressure measurements were not so reliable in the lateral position. This study concludes that clinical practitioners can obtain accurate pulmonary capillary wedge pressure measurements in post-operative cardiac surgical patients positioned in either the left or right 60 degrees lateral position. Further research is however required, with larger numbers from all sub-groups of the critical care population. Physiological and pathophysiological characteristics which preclude reliable pulmonary artery pressure measurements need to be identified. PMID- 8704391 TI - Funding for a national critical care nursing education research study. PMID- 8704392 TI - Australian Coordinating Committee on Organ Registries and Donation. PMID- 8704393 TI - The "Spanish Model"--an initiative aimed at increasing organ donation rates in Australia. PMID- 8704394 TI - Clinical research in nursing, why is it so hard? PMID- 8704395 TI - Mornington Peninsula Hospital Intensive care unit, Victoria, Australia. PMID- 8704396 TI - Update on contraception. 2: Post-coital and barrier methods. PMID- 8704397 TI - When should we refer children to the child psychiatry team? PMID- 8704398 TI - Special feeds for special infants: 1. The baby with a metabolic or allergic disorder. PMID- 8704399 TI - Children's dental development. 2: Six years old to early teens. PMID- 8704400 TI - Remedies for common family ailments: 8 vitamins and tonics. PMID- 8704401 TI - Deafness in childhood: 3. Cochlear implants: who, why and what are the results? AB - Some profound or totally deaf children cannot benefit from a hearing aid. In some cases a cochlear implant may be successful. The system compromises a small cable implanted into the inner ear, and externally-worn devices to collect and transmit the sounds to the ear. The cable consists of electrodes that respond to different frequencies of speech, imitating the action of the inner ear. It is not possible to predict the outcome in individual cases. At best, the child may subsequently be able to speak fluently and hear without lip-reading. At least, the child should be able to hear some background noise such as traffic, and be able to improve his or her lip-reading skills. It may take up to a year before improvements begin to show. The implant is intended to be permanent. Cochlear implants are still at a relatively early stage of development. Progress is cautious but the results so far are very encouraging. The entry gate is widening and the number of children being accepted for treatment is increasing. PMID- 8704402 TI - Skin care and nappy rash. PMID- 8704403 TI - Maternity policy. Caroline: a case of a pregnant teenager. AB - The government's health of the Nation strategy aims to reduce the incidence of conceptions among the under 16s by at least 50% by the year 2000. Teenagers need contraceptive services that they find approachable and easily accessible. Traditional family planning arrangements are not necessarily appropriate. Sex education in schools is sometimes too biological and does not relate to the teenager's personal situation and concerns. The main concern about teenage parents may be the long-term social rather than health consequences. PMID- 8704404 TI - "My child has asthma": some answers to parents' questions. AB - The incidence of asthma appears to have doubled since the 1970s. 15% of children in the UK are now estimated to have the condition. Children's asthma needs to be taken seriously by parents, health professionals and teachers. Its severity varies and it is a potentially life-threatening condition. The reason why children develop asthma is not known. Theories include a family tendency to asthma and allergy, house-dust mite and parental smoking. There is no clear evidence of a link between asthma and traffic fumes and air pollution. The vast majority of people with asthma take inhaled steroids, and these produce little or no side-effects. Tablet forms of the drug also have few side-effects if used for short courses of a few days. Only a minority need to take steroid tablets regularly, and need to discuss the issue of side-effects versus improvements in symptoms with their doctor. Most children use a preventer and a reliever medication. It is important that parents and child understand how and when to use each. Research suggests that two-thirds of children may grow out of their asthma symptoms, though asthma that is triggered by an allergic response may recur in later life. The prognosis for pre-school children is good. PMID- 8704405 TI - Quality assurance. Establishing a district-wide enuresis service. AB - When two authorities merged there was a need to unify and redefine two previously separate services. A steering group including the school nurse co-ordinator agreed the aims, objectives and standards for the new service. Clients (age range 5-18) see a CMO at the first visit and thereafter a school nurse. Rigorous audit of outcomes has enabled the service to demonstrate its effectiveness. PMID- 8704406 TI - Rubella vaccination: why vigilance must continue. PMID- 8704407 TI - Job-sharing: a recipe for success. PMID- 8704408 TI - Common foot problems. PMID- 8704409 TI - Hormone replacement therapy: women's need for information. PMID- 8704410 TI - In vitro fertilisation: a family made possible. PMID- 8704411 TI - Remedies for common family ailments: 10. Nasal decongestants. AB - Nasal congestion and sinusitis are common effects of cold and flu viruses. One treatment for nasal congestion is oral doses of vasoconstrictors such as ephedrine which mimic the action of the sympathetic nervous system. Care should be taken before these are prescribed or recommended as these drugs are contraindicated in patients on certain drugs or with certain conditions. Vasoconstrictors can also be given as nasal drops or sprays but for a limited period to avoid rebound congestion. Aromatic volatile oils, e.g. menthol, eucalyptus, can be given as rubs or inhalants (see manufacturers' instructions for use with young children). PMID- 8704412 TI - Hay fever: a problem on the increase. PMID- 8704413 TI - Child Safety Week: focusing on the local scene. PMID- 8704414 TI - Caring for the pregnant woman with sickle cell crisis. AB - Pregnancy poses particular risks for women with sickle cell disease and their unborn babies. There is an increased risk of spontaneous abortion and preterm labour. Painful sickle cell crises tend to occur more frequently during pregnancy. A district-wide protocol for dealing with crises would help ensure the woman receives prompt, effective, knowledgeable care from GPs, midwives and A&E staff. Careful assessment is essential and hospital admission is usually required. Adequate relief of pain, and sometimes blood transfusions, are the cornerstones of treatment. Health education plays a part in prevention of crises. Care could be improved by more specialist centres and specialist nurses or midwives. PMID- 8704415 TI - Management of diarrhoea by the primary health team. AB - For all age groups, fluid replacement with oral rehydration sachets is the mainstay of treatment. Antibiotic therapy will occasionally be needed where specific pathogens have been identified. Traveller's diarrhoea may be alleviated with antibiotic therapy. Attacks of viral gastroenteritis in the under-twos are common. Consistent advice from health visitors, practice nurses, midwives and the GP helps prevent parents from receiving conflicting messages. *In a baby, colic accompanied by diarrhoea and especially by blood in the stool, needs urgent referral to a doctor--it could be intussusception. Persistent diarrhoea should always be investigated. PMID- 8704416 TI - Breast milk for preterm infants. AB - The breast-feeding intentions and achievements of mothers of babies born prematurely were investigated in a prospective enquiry, a postal survey and by interview. The prospective inquiry was conducted from April 19 1993 to March 19 1994 on all 686 infants admitted to the two neonatal units in Nottingham. The aim was to see how many infants were offered mother's breast milk by any route. Feeding records were available for 593 infants of whom 64% were offered mother's breast milk. Gestation did not affect the mother's ability to produce milk. Infants of 24-28 weeks' gestation received breast milk for 4 to 5 weeks on average. The postal questionnaire was sent to 114 mothers whose infants were in the units for over seven days between April and October 1994. The aim was to learn how they felt about producing milk and feeding their preterm infant. 64 [59%] responded. 54% of responders and 49% of non-responders had intended to breast feed. Of the responders 65% produced some milk and 46% were home fully breast feeding. The home interviews were conducted 2-6 weeks after discharge with 16 mothers who fulfilled the criteria for the postal enquiry. The aim was to learn from mothers how the service might be improved and what motivated their choice of feeding. They requested clear guidance, agreed by all the professionals involved; a warm, comfortable and private environment; and a ready supply of breast pumps. Their choice of feeding was motivated primarily by previous experience and family pressure, not by advice from health-care professionals. CONCLUSIONS. Many mothers succeed in supplying milk for their preterm infants and many more might do so if the benefits of human milk to preterm infants were more actively advocated and the support service in the neonatal units developed. PMID- 8704417 TI - Safe not sorry: preventing baby and child accidents. PMID- 8704418 TI - Getting a job in general practice. AB - Practice nursing is unique in the NHS--one works with and for an employer (the GP) rather than an organisation. Before applying for a post the nurse should gain as much background knowledge as possible about working in a general practice. Both GP and nurse may have misconceptions or uncertainties about each other's role, e.g. the employer may have unrealistic expectations of nursing competencies and potential. The nurse may be unfamiliar with GPs' status as independent contractors, hence their freedom to set terms and conditions of service. Through a process of informal and formal interviews, both parties need to establish clearly what is understood and expected of the other. Study the terms and conditions of service and the final contract carefully before signing. Be open and honest about what the job has to offer and what one can offer the job. Then, once you have accepted, go forward positively! PMID- 8704419 TI - [Study on essence of liver-qi stagnation in ulcerative colitis]. AB - The links between stagnation of the Liver-Qi in the pathogenesis and ulcerative colitis (UC) were clinically and experimentally studied using the principle of nourishing the Liver in treated group and the principle of invigorating the Spleen in control group. The results showed that the effective rate was 96% in treated group and 82% in control group, and the difference was significant (P < 0.05); the formation rate of E rosettes and the transformation rate of lymphocytes were significantly raised in the two groups compared with the pre treatment period, treated group was evidently superior to control group in the rate of dysfunction of autonomous nerve system (P < 0.01). The level of intestinal styrenated phenol (SP) and vasoactive intestinal polypeptide (VIP) of UC model were determined in rats. The results showed that the level of SP and VIP significantly increased in UC model group. They markedly lowered in treated group compared with model group (P < 0.01) and there was significant difference in comparing with control group (P < 0.01). It revealed that Wei Chang Ning, a drug to nourishing the Liver, had the action of regulating neurological endocrinological (gastrointestinal hormone)-immunological system. PMID- 8704420 TI - [Clinical study on treatment of peptic ulcer with bushen kangkui decoction]. AB - The clinical controlled study of Bushen Kangkui Decoction (BSKKD) on 150 peptic ulcer patients showed that it had a similar effect to injectio cimetidine, and had a lower rate of relapse. And the long-term therapeutic effect was better than that of injectio cimetidine. Simultaneous RIA showed that PGE2 of most treated patients was below normal level before the treatment, but it has increased in various degree after the treatment. The patients under the treatment of BSKKD had significant difference in PGE2 level (P < 0.05), while those treated with injectio cimetidine didn't have (P > 0.05). So the BSKKD has an effect on the increase of PGE2 level of the peptic ulcer patients. PMID- 8704421 TI - [Studies on function of vegetative nervous system in cardiopathic patients with heart-qi deficiency syndromes and heart-yin deficiency syndromes]. AB - To explore the changes of the function of vegetative nervous system in the patients with the syndromes of Heart-Qi Deficiency (HQD) and Heart-Yin Deficiency (HYD), 53 cases of heart diseases were divided into HQD and HYD, the duration between heart beats, differences of blood pressure in the lying and standing position, and urine-catecholamine/24 hrs was determined separately, and compared with control group. The results showed that the duration between heart beats and heart rate in the lying position in those patients of HQD didn't manifest significant difference with control group. But in 78.9% of patients, the differences of heart rate in expiration/inspiration, surpassed 15 beats per minute, and the proportion of 30/15 was less than 1.03. Compared with control group, the difference was significant (P < 0.01). Meantime, the differences of heart rate of lying/standing position was less than 15 beats per minute, and the difference of blood pressure had an increasing tend in 47.4% of the cases. Those patients of HYD didn't show any difference in the above indexes. Urine catecholamine/24 hrs in both of the two groups surpassed that of control group (P < 0.01). In short, this study showed that syndromes of HQD and HYD both displayed disturbed function of the vegetative nervous system, and their types and severities were correlated with the difference of Syndromes. PMID- 8704422 TI - [Study on treatment of infectious shock with recipe of liqi huoxue and kaibi gutuo]. AB - Based on the pathogenesis of infectious shock which were stagnation of Qi, blood stasis, impairment of body resistance and trends to collapse, a series of new preparations of traditional Chinese Medicine was developed, Injection Kangjue Tongmai, Injection Yiqi Jiuyin and Injection Yiqi Huiyang, which could Liqi Huoxue and Kaibi Gutuo (regulate the flow of Qi, promote blood circulation, strengthen the body resistance and remit collapse), the result of 183 cases of infectious shock treated with them showed that the mortality was 4.37%, significantly lower than that of control group (23.0%). All three injections could rapidly elevate blood pressure and stabilize it for prolonged time, increase renal blood flow and lower blood viscosity. In animal experiment, the injections reduced lipid peroxide of vital organs, stabilized bio-membrane, protected the cell structure and maintained their normal functions. PMID- 8704423 TI - [Preliminary study on early fibrosis of chronic hepatitis B treated with Ginkgo biloba Composita]. AB - The purpose of this paper was to study the effect of Ginkgo biloba Composita (GBC) on liver collagen fibrosis. The 86 patients were suffering from chronic persistent and active hepatitis B with early fibrosis confirmed by liver biopsy. Serum pro-collagen-III-peptide (PIIIP), laminin (LN), superoxide dismutase (SOD), malonyldialdehyde (MDA) were determined and some patients' biopsies examined with light and electron microscope before and after treatment were observed. By the end of 3 months, the results showed that PIIIP, LN, SOD, MDA were significantly decreased after GBC treatment (P < 0.01) and liver fibrosis of patients was partly reabsorbed and had remission. It was suggested that GBC was effective in arresting the development of liver fibrosis of chronic hepatitis. PMID- 8704424 TI - [Clinical study on acute cerebral hemorrhage with zhong feng I, II oral liquor]. AB - The 120 patients with acute cerebral hemorrhage were divided into two groups: tested group and control group. The tested group were treated with Zhong Feng I oral liquor (acute stage), Zhong Feng II oral liquor (convalescent stage) and Western medicine, the control group were treated with Western medicine alone. The results showed that: the course of acute stage, and the time for restoring consciousness and the time for eliminating high intracranial pressure were shorter in tested group than those in control group; The mortality and rate of causing disability lowered. The rate of intracranial hemorrhage absorption was promoted in tested group (81.67%) than that in control group (58.37%), P < 0.01; The efficacy in various aspects was superior than that of control. PMID- 8704425 TI - [Relationship between anti-coagulation system changes of coronary heart disease patients and different syndrome-type in TCM]. AB - The level of Antithrombin-III : antibody (ATIII : A), Antithrombin III: antigen (ATIII : Ag), protein C antigen (PC : Ag), Total protein S : antigen (TPS : Ag) and Free protein S : antigen (FPS : Ag) were determined in 46 cases of coronary heart disease (CHD) and 40 cases normal control with the thrombin gelplaque technique and the immunoelectrophoresis assay. The results showed: the level of the PC: Ag and TPS: Ag of Qi stagnation type of CHD were significantly higher than those in normal control group (P < 0.05), the level of the AIII: Ag, ATIII : Ag in blood stasis type of CHD were significantly less than those in control (P < 0.05). These suggested that the level of PC: Ag and TPS: Ag might be used as referential indices of the Qi stagnation type; the level of ATIII: A, ATIII : Ag might be used as those of blood stasis type in CHD. PMID- 8704426 TI - [Effect of zhuchun pill on immunity and endocrine function of elderly with kidney yang deficiency]. AB - After randomly dividing 45 elderly males with Kidney-Yang Deficiency into Zhuchun Pill (ZCP) group and placebo (starch capsule) group by single blind method, and treating them for 3 months, we found that ZCP could greatly improve the main symptoms of Kidney-Yang Deficiency, P < 0.01 by comparison with control group: the 3H-TdR lymphocyte transformation (3H-TdR LCT) rate, level of serum C3, IgA, IgD, IgG & IgM went up (P < 0.01); contents of plasma adrenocortical hormone (ACTH), testosterone (T) and cyclic adenosine monophosphate (cAMP) increased (P < 0.01-0.001): the level of superoxide dismutase (SOD) in serum also increased while lipid peroxide (LPO) in plasma decreased (P < 0.01). But for control group, there were no remarkable changes after treatment, so it revealed that the improvement of Kidney-Yang Deficiency by ZCP is realized through regulating the organism's immunity and endocrine function as well as through scavenging the free radical agents. PMID- 8704427 TI - [Ginkgolides antagonizing some effects of platelet-activating factor in vitro]. AB - The mixture of platelet-activating factor (PAF) and platelets produced significant contraction of guinea pigs' bronchus, while the contraction induced by PAF alone was mild relatively, the IC50 were 6.14 x 10(-7) mol/L and 6.32 x 10(-4) mol/L respectively. There was significant difference between these two groups (P < 0.05). When the platelets were pre-incubated with ginkgolides for 10 minutes in Tris-Tyrode's buffered saline, effects of the PAF and platelets mixture were significantly inhibited (P < 0.01). Exposure of guinea pigs' bronchus to PAF in vitro resulted in a loss of beta-adrenergic receptors and responses to isoproterenol, and this effect of PAF was prevented by prior incubation of the guinea pigs' bronchus with ginkgolides (P < 0.05). The results showed ginkgolides were a potent PAF antagonist. PMID- 8704428 TI - [Effect of ligustrazine on kanamycin ototoxicity]. AB - By measuring cochlear superoxide dismutase (SOD) activity, malondialdehyde (MDA) and electrocochleogram (ECoG) in guinea pigs, the antioxidation of Ligustrazine on Kanamycin (KM) was studied. The results showed that SOD activity, MDA content and threshold value of ECoG were significantly increased after KM was administered, while it markedly reduced after Ligustrazine was given. It was considered that lipid hyperoxidative reaction was correlated with KM ototoxicity, Ligustrazine could prevent and treat KM ototoxicity by antioxidation. PMID- 8704429 TI - [Protective effect of "wei tong ling" on gastric mucosa and influence on quality of gastric ulcer healing]. AB - The acute gastric ulcer rat models were induced by dehydrated alcohol, 0.6 N hydrochloric acid and 0.6 N sodium hydroxide, and the chronic gastric ulcer rat models were established by means of acetic acid, the protective effect of Chinese medicine "Wei Tong Ling" (WTL) on gastric mucous membrane was studied. Using histochemical mucin stain, AgNOR stain and immunohistochemical technique the regenerated mucosa of healed gastric ulcer induced by acetic acid in rats was observed quantitatively. They were compared with that of WTL. The results showed that the regenerated mucosa of healed gastric ulcer might be the morphological basis for the recurrence of gastric ulcer and be associated with canceration. WTL could not only accelerate the healing of ulcer but also raise the quality of gastric ulcerous healing which was beneficial for the prevention of ulcer recurrence and canceration. The protective effect of WTL on gastric mucosa was confirmed by various assays. PMID- 8704430 TI - [Effect of acupuncture on immunologic function and histopathology of transplanted mammary cancer in mice]. AB - This experiment mainly describes the effects of acupuncture on immunologic function and histopathology of transplanted mammary cancer in mice. The results were as follows: in acupuncture group, NK cell activity and T-lymphocyte positive rate of acid alpha-naphthyl acetate esterase (ANAE) and lymphocyte transformation rate were all increased. Compared with the control group, there was a significant difference (P < 0.01). The difference was insignificant, when compared with normal group (P > 0.05). Comparing the pathology grading of acupuncture group with control group, it showed marked difference in pathological section (P < 0.01). Adenoid structure and the degree of lymphocytic infiltration also have marked difference between acupuncture and control group (P < 0.05). Less tumour volume in acupuncture than control group were observed (P < 0.01). This indicated that acupuncture might increase the immunologic function of transplanted mammary cancer in mice and inhibit the growth of mammary cancer and enhance both differentiation level of mammary cancer cells and lymphocytic infiltration. Possibly acupuncture mightt reduce the malignancy of mammary cancer cells. PMID- 8704431 TI - [Characteristics of CO2 expiration of skin on limb segment of large intestine and stomach meridian]. PMID- 8704432 TI - [Effects of traditional Chinese medicine in chemotherapy of acute leukemia]. PMID- 8704433 TI - [Exploration of clinical study of antileukemia cell drug-resistance by traditional Chinese medicine]. PMID- 8704434 TI - [Recent advances in experimental and clinical study on ligustrazine in treating respiratory disease]. PMID- 8704435 TI - [Effect of acupuncturing houxi and shenmen in treating cerebral traumatic dementia]. AB - Sixty-two cerebral traumatic dementia patients were randomly divided into two groups: acupuncture group (32 cases) and physiotherapy group (30 cases). The changes of mini-mental status examination (MMSE) scores and auditory evoked potential P300 were observed before and after treatment. RESULTS: marked effective rate and effective rate of acupuncture group were 46.9% and 81.3% respectively, significantly higher than 10.0% and 30.0% of physiotherapy group (P < 0.01). It was found that after treatment the above-mentioned indexes in acupuncture group changed significantly (P < 0.05 - 0.001), while in physiotherapy group were not (P > 0.05). The therapeutic effect in acupuncture group was better than that in physiotherapy group (P < 0.001). The results confirmed that the cognitive function of cerebral traumatic dementia patients was enhanced effectively with acupuncturing at Houxi and Shemen. PMID- 8704436 TI - [Study on relationship between nail-fold microcirculation and histopathology of patients with vocal cord polyp and blood stasis syndrome]. AB - Nail-fold microcirculation of 50 vocal cord polyp patients was observed and compared with hoarseness patients without vocal cord polyp. The results showed that peripheral microcirculation was disturbed in the vocal cord polyp patients. Histopathologic observation of 200 vocal cord polyp patients was made. It was found that principal pathologic change concentrated on the lamina propria mucosae of vocal cord and the most prominent manifestation was disturbance of topical microcirculation of Reinke's layer. In microcirculation aspect, the relationship between polyp of vocal cord and the Blood Stasis Syndrome was confirmed, and pathogenic mechanism of vocal cord polyp was discussed. PMID- 8704437 TI - [Relationship between thymectomy and syndrome differentiation-typing treatment of myasthenia gravis]. AB - According to the theory of traditional Chinese medicine, 37 patients suffering from myasthenia gravis were Syndrome-differentiated into following types preoperationally: (1) Both Qi-Yin Deficiency of Kidney-Spleen, 21 cases. (2) Yang Deficiency of Kidney-Spleen, 15 cases. (3) Qi Deficiency of Spleen only, one case. Thymectomy were done in all cases. Among them, 19 cases had thymoma. The results showed the relationship between the types and the resectionability rate and post-operative crisis rate. On the first two types, they were 95.2% and 9.5%, 46.7% and 73.3% respectively. It was considered that Syndrome differentiation typing in TCM in the myasthenia gravis patients would provide an positive and reliable index for predicting the resectionability and prognosis of the patient. PMID- 8704438 TI - [Clinical and experimental study of yangxueshengru oral liquor in promoting puerperal breast milk secretion]. AB - Applying Yangxueshengru Oral Liquor (YXSR) to treat 103 cases of puerperal insufficient lactation. Six hours after delivery, the parturient took YXSR, its effective rate was 98.06%. The results revealed that it could improve breast milk secretion significantly and increase daily lactation amount of parturient, insufficient lactation rate was reduced. The content of protein and trace elements (Fe, Zn and Mn, etc) in colostrum increased, the difference between medicated group and control group was significant (P < 0.05-0.001). Animal experiment showed that YXSR could increase the response of prolactin (PRL) level of model rats (P < 0.01), the serum, PRL and cortisol level of the rats were also raised significantly (P < 0.01). PMID- 8704440 TI - [Effect of acupuncture on heart rate variability in coronary heart disease patients]. AB - The heart rate variability (HRV) of 20 coronary heart disease patients has been determined before and after acupuncture treatment by using frequency domain analysis of HRV. The results showed that there was significant difference between the change of low frequency (LF) components in HRV before and after manipulatory acupuncturing Neiguan group and electro-acupuncturing Neiguan group (P < 0.05), while there was no significant difference in the control group (P > 0.05), also no significant difference on the change of high frequency (HF) components in HRV before and after acupuncture in both groups. In the electro-puncturing group, the LF/HF change was marked (P < 0.05). And in manipulatory acupuncturing group, the value of LF reduced to the lowest level 10 minutes after acupuncture treatment, and then gradually restored 20 to 30 minutes later, without any rebound phenomenon. It suggested that acupuncture could regulate and improve HRV in coronary heart disease patients. Its mechanism might be relevant to the central regulation and neurotransmitter participation. PMID- 8704439 TI - [Clinical and laboratory studies of effect of longevity-antihypertensive-mixture on elderly hypertension with kidney deficiency]. AB - Longevity-Antihypertensive-Mixture (LAM) is a Chinese herbal drug, which could tonify Kidney in treating the hypertension with Kidney Deficiency in aged patients. RESULTS: 302 patients were randomly divided into three group: The treated group was subdivided into group I (107 Kidney Yin Deficiency patients) and group II (96 non-Kidney Yin Deficiency cases) and group III (99, control group). After treatment of LAM for 8 weeks, the rate of normalizing the blood pressure was 92.52% in group I while in group II, it was 84.43%, the symptoms were also significantly alleviated, and in group III, the rate was 70.71% only. The aorta compliance in group I significantly improved after LAM treatment (P < 0.025), the quality index of left ventricle reduced in treated group, and there was significant difference between treated and control groups. After treatment, LAM could shorten the length of extracorporal thrombus in group. The study showed that LAM could treat hypertension through LAM, administration with good effect. PMID- 8704441 TI - [Experimental study on the effect of abstinence with herbal preparation composite dong yuan gao]. AB - The effect of herbal preparation "Composite Dong Yuan Gao" (CDYG) on the animal model of drug dependence was studied. After the morphinistic models of white rats and mice were made by ever increasing doses of morphine each time, the animals were divided into large dose CDYG group, small dose CDYG group, sustained morphine group, and control group. The withdrawal symptoms were observed after naloxone was given to the animals intraperitoneally. The results showed that in the CDYG groups withdrawal symptoms of the addicted white rats were alleviated, the number of jumps of the addicted mice was reduced, the scores of the principal symptoms of the animals were decreased, while the body weights of the animals were gained. Comparing with the control group, the difference was significant (P < 0.01). The effects of the CDYG enhanced in the large dose group, which suggests that the CDYG has abstinence effects. PMID- 8704442 TI - [Ia antigen expression of peritoneal macrophage in mice model of yang deficiency and the effect of aconitine]. AB - The immune associated (Ia) antigen expression (IAAE) of macrophage is one of the important indicators of the specific immunity in the body. In this paper, mice of Yang Deficiency model was made by injecting corticosterone in the hind legs of mice to inhibit the function of hypothalamus -pituitary-adrenal axis. Authors observed the changes of IAAE in model mice and the effect of Aconitine on it. The results indicated that: (1) In model mice, the IFN-gamma induced IAAE of peritoneal macrophages was inhibited. The difference of IAAE between the model and the normal mice was significant (P < 0.01). (2) Aconitine could significantly increase the IAAE of model mice (P < 0.01). These data implied that the immuno suppression in the model mice of Yang Deficiency might be due to the lowered IAAE of macrophages under this condition. The promoting effects of Aconitine on IAAE enhanced the antigen presenting ability of macrophages and the immune response of the model mice. It might be related to the therapeutic mechanism of Aconitine on Yang Deficiency Syndrome. PMID- 8704443 TI - [Effects of osthol and total-coumarins from Cnidium monnieri on immunological function in kidney yang deficiency mice]. AB - Mice were injected intraperitoneally with hydrocortisone acetate to replicate the animal model of Kidney Yang Deficiency (KYD). Osthol and total-coumarins (TCR) from the fruit of Cnidium monnieri were administered orally to model mice. The effects of osthol and TCR on the immunological function of the KYD mice were observed. The results showed that compare with those in normal mice, the percentage of phagocytosis and the index of macrophage phagocytosis, the level of serum hemolysin and the proliferation of lymphocytes lowered significantly (P < 0.01) in model mice. It was shown that administering osthol or TCR could notably (P < 0.01) prevent the above-mentioned lowering in model mice, revealing that osthol and TCR could improve the immunological function of KYD mice. PMID- 8704444 TI - [Progress in the treatment of opiate dependence]. PMID- 8704446 TI - UKCC: is it now fit to govern? PMID- 8704445 TI - Nurses' pay award lacks vision. PMID- 8704447 TI - Preoperative visiting: a role for theatre nurses. AB - Over the past 20 years, giving preoperative information to patients has been recognized as benefiting them greatly (Hayward, 1975; Boore, 1978). Similarly, preoperative visits by theatre nurses have been widely advocated to allay patient anxiety. This article discusses preoperative visiting from the perspective of its value to theatre nurses, particularly in the planning of patients' nursing care and in the application of nursing models and a team approach to the delivery of care. In so doing, preoperative visiting may be a life-line for theatre nurses at a time when their unique contribution within the operating theatre is increasingly under scrutiny (Bevan, 1989). PMID- 8704448 TI - Treatment of venous leg ulcers: 1. AB - The management of venous leg ulcers is an important issue for many health-care professionals. If treatment is to be successful, it is vital that clinicians understand the physiology and disease processes of the venous system in general, and of venous ulceration in particular. Part one of this two-part article presents an outline of the normal physiology of the venous system in the lower limb followed by a brief history of venous leg ulcers and an in-depth analysis of the way in which venous disease can lead to ulceration. The analysis compares traditional theories of leg ulcer development with current hypotheses. Part two will look at the treatment of venous ulcers and associated nursing implications. PMID- 8704449 TI - The biology of Alzheimer's disease: 2. AB - In this article, the second of two parts, the authors continue their review of the biological processes of Alzheimer's disease and the implications for nursing practice. A glossary of terms can be found in the first article (Vol 5(3): 162 7). This article considers the function of the subcortical area of the brain and explores its effects on the world of the person with Alzheimer's disease. Nursing responses are also discussed. PMID- 8704450 TI - Brighter futures: focusing on sheltered housing and tenants with dementia. PMID- 8704451 TI - Nurses' responses to patient behaviours that threaten health. PMID- 8704452 TI - Smoking with patients: policy us therapy. AB - While it is largely accepted that smoking is injurious to physical health, there is no general consensus on the value, or otherwise, of mental health nurses smoking in the company of their patients in the creation and maintenance of therapeutic relationships. This survey of key opinion-formers within medium- and high-security mental health-care facilities was designed to ascertain whether a consensus exists. PMID- 8704453 TI - Effects of cannabis abuse on people with serious mental health problems. AB - There is clear evidence of illicit drug abuse in psychiatric hospitals (Deahl, 1991), and illicit drug use has been associated with relapse in people with serious mental health problems. Cannabis abuse in the general population is common and has both desirable and undesirable effects on users. However, when cannabis is used by people with serious mental health problems, a more pronounced effect may be seen. There is little evidence to suggest that cannabis 'causes' schizophrenia, but its abuse may prove to be a trigger in vulnerable individuals. Studies have demonstrated that cannabis abuse can exacerbate the symptoms of schizophrenia and may reduce the effectiveness of antipsychotic drugs. Mental health nurses need to develop interventions in order to manage illicit drug abuse effectively in both the hospital and community settings. PMID- 8704454 TI - Is it ever permissible to discharge an illicit drug user from hospital? AB - Is it ever permissible to discharge a patient who uses illicit drugs on hospital premises? It is argued that there is nothing inherent in the use of illicit drugs by a patient which justifies his/her discharge. However, the implications of the drug use need to be examined. First, if a patient's activity presents a significant risk to others, the hospital may be in breach of safety regulations and may need to secure alternative provision of treatment. Second, if the continued use of drugs limits the benefits of treatment, this treatment may be considered an inappropriate use of resources. PMID- 8704455 TI - Sexual dysfunction following radiotherapy for cervical cancer. AB - The majority of patients with cervical cancer are treated with radiotherapy, which has many potential side-effects. This article reviews the literature, emanating primarily from the USA, which seeks to determine why sexual dysfunction is so often a problem for patients following such treatment and why nurses appear to be so reluctant to deal with this issue. The exact nature and severity of the difficulties experienced by women will be examined and also the reasons why nurses appear so reluctant to deal with this issue. Finally, possible solutions and strategies for action will be suggested. PMID- 8704456 TI - Catheters: design, selection and management. AB - Urinary catheterization is a common procedure that is performed on 12% of hospital patients (Crow et al, 1996) and 4% of community patients (Getliffe, 1990). When considering catheterization as a method of care, many decisions need to be made in order to select the optimum equipment and provide effective catheter care. This includes selecting the type of catheterization that needs to be performed, i.e. intermittent or indwelling, and choosing the right catheter material, size and balloon infill volume. The type and design features of the urinary drainage and suspensory system have a direct effect on individual patient comfort, dignity and life. Aspects of catheter care that need to be considered include: meatal hygiene, fluid intake, bladder washouts, constipation, clamping catheters, collecting specimens of urine, fixation of catheters and sexuality. PMID- 8704457 TI - Causality and control: key to the experiment. AB - The main aim of the experimental approach in research is to discover causal relationships between variables. This article describes the concept of causality and discusses common and important aspects and factors related to causality, manipulation and control. The four principles which underpin the inference of causality in experiments are highlighted and the role of 'control' in achieving the aim of experimental studies is discussed. Some methods of control such as manipulation, randomization, matching subjects, and holding extraneous variables constant are considered. Experimental designs are underpinned by these concepts. Indication is given of the threats to internal validity, which will be discussed in detail in the next article in this series. PMID- 8704458 TI - Setting new ethical standards for practice is a collaborative process. PMID- 8704459 TI - A&E services need a major overhaul. PMID- 8704460 TI - Pre-admission education for patients undergoing cardiac surgery. AB - Traditionally, preoperative information has been provided to patients on admission, the day before surgery. This was thought not to be the ideal time. An exploratory pilot study was undertaken to evaluate a new information service for patients about to undergo cardiac surgery. Questionnaires were administered to 20 patients who had attended a pre-admission education programme and 20 patients who had received information from the ward staff on admission. The aim was to assess whether patients fears and anxieties were reduced by the provision or pre operative information before admission for surgery. One hundred percent of the patients felt that they benefited from the pre-admission programme and 76% felt that their anxieties had been relieved. PMID- 8704461 TI - Infection control in GP surgeries: safe practices? AB - Any increase in the use of invasive procedures carries with it an increased risk of cross-infection. The spread of the human immunodeficiency virus, together with increasing awareness of hepatitis B virus, cytomegalovirus and human papillomavirus have focused attention on the nature and adequacy of procedures to prevent cross-infection in general practice. PMID- 8704463 TI - Continence following childbirth. AB - Urinary and faecal incontinence are problems that can severely limit the social, emotional and sexual being of the woman. Childbirth is one of the major causes of incontinence in women, especially following long second stages of labour, traumatic deliveries, large babies and in multigravidae. This article highlights the issues surrounding childbirth and incontinence and the implications that they have for nursing care. Nurses, especially community nurses, midwives and practice nurses, need to take a proactive stance to the identification of incontinence in women. Once a problem has been identified, it is important that the woman has a thorough assessment and examination before deciding on a plan of care. The use of pelvic floor exercises is the main non-surgical treatment for women with mild to moderate stress incontinence and these can be taught by nursing staff. PMID- 8704462 TI - Implementation of an aspect of self-care. AB - This article describes how a nursing development unit (NDU), through the evolution of a self-care philosophy, responded to the changing needs and demands of patients. Patients in the NDU appeared to want to be more informed and involved in their health care. In addition, decreased length of stay as a result of increased sophistication of medical techniques and the need to have a faster throughput of patients has meant that a philosophy is required which is focused around empowering patients to take ownership for their own health care. The application of this philosophy is demonstrated through a case study which describes the experience of a gentleman who was cared for within the NDU. PMID- 8704464 TI - The changing role of the continence adviser. AB - The role of the continence adviser is evolving and developing within today's health service. Research evidence is available to define the role and its principal functions. Since the inception of the role in the early 1980s, there is still wide variation in interpretation, lack of uniformity and wide variance of service provision across the country. It is time to remove the title of continence adviser as it no longer portrays the role as it has developed. In addition, the taboo nature of the word, incontinence, has to be considered as a potential barrier to progress and development. An ENB course has now been established to ensure that continence advisers can take their rightful place alongside their colleagues in other specialist areas of nursing. As these develop, aspects of the continence adviser's role may be under threat. Continence advisers will therefore have to sell their services in the marketplace and move away from pad-oriented activities. PMID- 8704465 TI - Treatment of continence in people with learning disabilities: 1. AB - This article is the first of a four-part series which examines the continence based needs of people with a learning disability. It considers the difficulty in defining learning disability as it is not a unitary phenomenon but covers a spectrum where learning delay is present to varying degrees. At best, the term is used administratively and therefore people with learning disabilities are those who are known to agencies providing services for such people. However, less than half of the total UK learning disabled population will not be identified as suffering impaired intellectual ability. So, even as an administrative term, learning disability fails. PMID- 8704466 TI - Understanding stress in the ICU setting. AB - This article discusses the physiological effects of stress on patients, relatives and staff in the intensive care unit (ICU). It examines the short- and long-term effects of cortisol on various systems in the body. The information contained in this article encompasses the recognition and understanding of the general adaptation syndrome as described by Seyle (1975). Many nurses working in ICU may identify with the symptoms and subsequent physiological and psychological responses to stress and thus enable informed discussion when planning the care of their patients or support of the relatives, colleagues and themselves. PMID- 8704467 TI - Causality and control: threats to internal validity. AB - The essence of experimental research is to establish causal relationships between variables and this requires internal validity. An experiment is said to have internal validity when extraneous variables have been controlled. However, there are many threats to internal validity. This article considers 13 of them, providing examples which clarify and help to identify under what conditions these threats may occur. The threats discussed are history, maturation, testing, instrumentation, statistical regression, selection mortality, interactions with selection ambiguity about the direction of causal influence, diffusion of imitation of treatments, compensatory equalization of treatments, compensatory rivalry by respondents receiving less desirable treatments and resentful demoralization of respondents receiving less desirable treatments. PMID- 8704468 TI - Autonomy and paternalism: partners or rivals? AB - This article is concerned with the ethical relationship between autonomy and paternalism. In the health care setting both clients and professionals can be profoundly affected by the way in which this relationship is interpreted and this article sets out to explore this phenomenon. It began by offering some well established definitions of autonomy and paternalism and goes on to examine the two principles in detail as they relate to persons in good health and to those in ill health. Finally an attempt made to answer the question of whether the two concepts are in conflict or partnership. PMID- 8704469 TI - Clinical supervision: what do you do after saying hello? PMID- 8704470 TI - Nurses must not become the mechanical hands of doctors. PMID- 8704471 TI - The role of the nurse in preventing intensive care psychosis. PMID- 8704472 TI - Doing nursing praxis research in New Zealand. PMID- 8704473 TI - Breastfeeding and paid employment in New Zealand: what role for nurses? PMID- 8704474 TI - Akenei Hei--pioneer and nurse leader. PMID- 8704475 TI - George's death--understood through Swanson's theory of caring. PMID- 8704476 TI - Moving from an oral tradition: the Praxis story. 1987. PMID- 8704477 TI - Staffing rural hospitals. Strategies for survival. PMID- 8704478 TI - Firearms public safety. PMID- 8704479 TI - Women family physicians and rural medicine. Can the grass be greener in the country. PMID- 8704480 TI - Benefits of exercise overlooked? PMID- 8704481 TI - Equal health care for men and women. PMID- 8704482 TI - Controversial use of bromocriptine. PMID- 8704483 TI - Use of the new quinolones during pregnancy. PMID- 8704484 TI - Dermacase. Fixed drug eruption. PMID- 8704485 TI - Each of the following has been shown to be effective for cyclical breast pain. PMID- 8704486 TI - Causes of horse-related injuries in a rural western community. AB - OBJECTIVE: To determine the causes of horse-related injuries in a rural western community. DESIGN: Prospective identification of persons with horse-related injuries and retrospective interviews with patients or witnesses to determine causes. SETTING: A small rural community in Alberta where the western style of riding predominates. PATIENTS: All patients presenting to two family medicine clinics or to the Sundre General Hospital emergency department. MAIN OUTCOME MEASURES: Factors contributing to the injury as recalled by patients or witnesses, and characteristics of the persons, horses, and injuries. RESULTS: Two thirds of the 150 injuries were caused primarily by horses and one third primarily by patients risk taking or inattention. The most common horse behaviour that caused injuries was "spooking," but several other behaviours also were identified as primary causes. The injuries were varied and relatively severe. Only one person was wearing a helmet. CONCLUSIONS: Horse-related injuries often are caused by characteristic horse behaviours. PMID- 8704487 TI - Training for rural practice. Are graduates of a UBC program well prepared? AB - OBJECTIVE: To evaluate preparedness for rural practice and to ascertain where graduates of a community-based rural training program practise. DESIGN: Mailed cross-sectional survey. SETTING: Rural communities in British Columbia. PARTICIPANTS: Graduates of the University of British Columbia's (UBC) rural training program from 1982 to 1991 and a random sample of non-program-trained rural BC physicians. MAIN OUTCOME MEASURES: Self-reported preparedness for rural practice in various areas of family medicine and in aspects of professional and personal life in rural settings. Locations of practice. RESULTS: Rural program graduates reported themselves better prepared in family medicine, community medicine, practice management, and behavioural science. Non-program-trained rural physicians thought themselves better prepared in medical subspecialties. Responses in pediatrics, obstetrics and gynecology, and surgical preparation showed no important differences. Rural program residents were located in rural areas (51%), regional settings (20.5%), and metropolitan areas (17.9%). CONCLUSION: Graduates of the UBC rural training program consider themselves better prepared for rural family practice than non-program-trained rural physicians in several areas of family practice. Most graduates of the program were practising in rural and regional settings. PMID- 8704488 TI - Chest pain in family practice. Diagnosis and long-term outcome in a community setting. AB - OBJECTIVE: To describe diagnostic distribution and outcome of chest pain among patients attending an urban family practice. DESIGN: Retrospective, descriptive chart review. SETTING: Primary care practice. PARTICIPANTS: All patients contacts for chest pain at Fossvogur Health Centre in the years 1989 and 1990 (193 contacts with 189 patients) were examined. One patient died before follow up and two could not be reached for follow up; they were excluded from the study. Of the 190 contacts and 186 patients studied, one patient who had two contacts with the clinic died during the study. MAIN OUTCOME MEASURES: Age and sex distribution, physical examination, investigations, diagnosis, and treatment; well-being of every patient was checked 3 to 4 years after initial contact. We asked about evolution of symptoms and looked for possible misdiagnosis. RESULTS: Musculoskeletal pain was diagnosed in 48.9% of contacts, heart diseases in 17.9% and 9.5% had undiagnosed chest pain. The history was the main diagnostic tool for patients with musculoskeletal diseases, while patients with heart diseases were examined more carefully and underwent more diagnostic procedures. Follow up showed that no serious disease had been missed in spite of restrictive use of laboratory investigations. CONCLUSIONS: The working methods of family doctors who examined patients with chest pain in this health centre can differentiate between patients with serious diseases and those with benign conditions. PMID- 8704489 TI - Postgraduate training for rural family practice. Goals and opportunities. AB - PROBLEM BEING ADDRESSED: The continuing shortage of rural family physicians in Canada. PURPOSE OF PROGRAM: To further develop training for rural family practice so that adequate numbers of rural family physicians will be appropriately prepared. MAIN COMPONENTS OF PROGRAM: All family medicine residents should have the opportunity to experience the joys and challenges of rural family practice. Rural family medicine training streams provide the best education for family medicine residents who are planning a career in rural family medicine. Integrated training for rural family practice should be high-quality, academically sound, needs-driven, evidence-based, learner-centered, and outcome-measured. This involves comprehensive development of curricula that provide specific skills and appropriate core subjects in rural practice as well as a solid family medicine foundation. contextual and experiential learning in areas similar to or in actual areas where there is a need for rural physicians, and appropriate hospital rotations to learn skills for the hospital role of many rural family doctors, are important components of rural family medicine training. CONCLUSIONS: Postgraduate rural family medicine training programs can be further focused and developed to train more physicians with the knowledge, skills, and attitudes required for rural practice. PMID- 8704490 TI - Glucose monitoring as a guide to diabetes management. Critical subject review. AB - PURPOSE: To encourage a balanced approach to blood glucose monitoring in diabetes by a critical review of the history, power and cost of glucose testing. DATA SOURCES: The Cambridge Data Base was searched and was supplemented by a random review of other relevant sources, including textbooks, company pamphlets, and laboratory manuals. STUDY SELECTION: Keywords used were "glucosuria diagnosis," "blood glucose self-monitoring," "glycosylated hemoglobin," and "fructosamine" for the 10-year period ending 1992, restricted to English language and human. DATA EXTRACTION: About 200 titles were retrieved and reviewed according to the author's judgment of relevance. FINDINGS: "Snapshot tests" (venous and capillary blood glucose) and "memory tests" (urine glucose, glycated hemoglobin fractions and fructosamine) must be employed according to individual patients treatment goals. Day-to-day metabolic guidance is facilitated by capillary blood glucose testing for patients receiving insulin and by urine glucose testing for others. Capillary blood glucose testing is mandatory in cases of hypoglycemia unawareness (inability to sense hypoglycemia because of neuropathy) but is not a substitute for a knowledge of clinical hypoglycemia self-care. Criteria by reason (clinical judgement and cost effectiveness) must be separated from criteria by emotion (preoccupation with technology and marketing). No randomized studies show that any of these tests consistently improve clinical outcome. Optimal metabolic control and cost savings can be expected from a rational selection of tests. PMID- 8704491 TI - Chest pain in children. AB - Chest pain is usually a benign symptom in children. The most common identifiable causes are musculoskeletal. Often, no cause can be identified. Cardiac disorders are uncommon causes of chest pain children. Most causes can be diagnosed from history and physical examination. Treatment should be directed at the underlying cause. For idiopathic chest pain, reassurance and regular follow-up examinations are important. PMID- 8704493 TI - Policy on training for rural practice. PMID- 8704492 TI - Obsessive-compulsive disorder. Diagnosis and management. AB - We present three diagnostic tools to identify overt compulsive rituals, obsessional thinking, and neutralizing behaviours in obsessive-compulsive disorder (OCD) and describe the most effective cognitive-behavioural technique for treating obsessional thinking without overt rituals. Basic dysfunctional beliefs that lead to OCD are explained and integrated in the treatment model. We suggest how combined therapy can be used to tread OCD. PMID- 8704494 TI - Up front with backs. PMID- 8704495 TI - Certification Examination of the College of Family Physicians of Canada. Part 2. Conduct and general performance. PMID- 8704497 TI - Health care quality report cards receive grade--incomplete. PMID- 8704496 TI - Utilizing quality assurance as a tool for reducing the risk of nosocomial ventilator-associated pneumonia. AB - A multidisciplinary group was formed to develop strategies to reduce ventilator associated lower respiratory tract infections (LRI) in an intensive care unit (ICU) of a 540-bed acute care teaching medical center. The group process was facilitated by the Infection Director and the quality management specialist. The group was made of medical, nursing, and respiratory therapy staff. Quality improvement techniques were used to define the process of care for ventilated patients in the ICU. "State of the art" care was defined after a literature review and brainstorming sessions. Current practice and new concepts were then forged into a realistic protocol for the ICU. The resulting protocol was introduced into the ICU in May 1992. The information was communicated to Respiratory Therapy and ICU staff in writing and at department meetings. After a 4-month introductory period and learning curve process, a decrease in the endemic rate of ventilator-associated LRI was reduced in the fourth quarter of 1992. The mean 1993 LRI rate was 21 LRI/1000 ventilator days versus 26 LRI/1000 ventilator days in 1992 before protocol implementation. This equates to 18 LRI prevented in 1993. This represents a savings of $126,000. There was a significant decrease in the process variation of the monthly mean LRI/1000 ventilator days in 1993 from 1992. To date, there continues to be improvement with a mean of 16 LRI/1000 ventilator days reported in 1994. The ICU staff developed a multidisciplinary process evaluation and monitored staff implementation of the protocol over time. The results of the evaluation were used as feedback to measure protocol implementation. This was found to improve compliance with the protocol. Both the process (care of ventilated patients) and the outcome (number of LRI) have been improved through use of continuous quality improvement concepts and transdisciplinary interventions. PMID- 8704498 TI - Developing an outcomes measurement system: the value of testing. AB - Outcomes measurement systems often experience similar challenges to implementing data collection and demonstrating value. This article examines the experiences of 451 hospitals participating in a 2-year research effort designed to assess the ability of participants to successfully implement collection of obstetric and peri-operative indicator data for development of an outcomes measurement system. Measures of ability to implement include rate of attrition and reasons for withdrawal, resources expended, months of data transmitted, internal factors that affected ability to operationalize data collection, and assessment of value of participating in the testing process. The findings indicate considerable variation in implementation ability, challenges encountered, and satisfaction with the experience. Several changes in the operational system were made in response to the findings. Many of the lessons learned from the testing experience may be applicable to sponsors of and participants in other outcomes measurement systems. PMID- 8704499 TI - Transfusion-to-cross-match community comparison data. AB - With blood supplies at low levels, and replacement and utilization becoming important issues throughout the nation, the Oregon Medical Professional Review Organization (OMPRO) looked at the blood replacement process in Oregon. The study addressed the utilization of type-and-screening and cross-matching in the acute care hospital setting-comparing the more common and expensive cross-match procedure to the less expensive and less utilized type-and-screen. The methodology involved two phases: (a) data collection, evaluation, and feedback to the hospitals and (b) post-monitoring. Although the study did not control for all possible variables, the post-monitoring results demonstrated an increase in the ratio from 0.569 (1992 data) to 0.577. For 1994, the total statewide transfusion cost was estimated at $5,909,700 as compared to $6,423,900 (1992 data)--a savings of more than $500,000 per year. By implementing minimal changes in the way blood is ordered and tested, hospitals can improve quality by maintaining a readily available blood supply, eliminating waste and saving money. PMID- 8704500 TI - Patients' perceptions of office medical practice: judging quality through the patients' eyes. AB - With increasing pressure to measure quality, patient-based assessments of medical care are becoming increasingly important. Patients offer a unique perspective for evaluating the nontechnical aspects of medical care. This study reviews the importance of utilizing patients' perceptions to measure quality of care in office settings. It also reviews the principles required to conduct a well done survey. The concept of patient perceptions differs from the more commonly measured concept of patient satisfaction, in that perceptions measure whether a patients' needs and expectations are met, in addition to satisfaction. One of the most accurate and efficient means of measuring patients' perceptions is through the use of surveys. As with all standardized data collection, creating and performing high quality surveys of patients' perceptions can be challenging. Valid and reliable patient survey data can enable practitioners to identify areas for improvement, and demonstrate to external reviewers the quality of care they provide to their patients. PMID- 8704501 TI - Assessment of delivery of preventive health services. AB - Our objective was to design and evaluate an instrument to assess the performance of a clinic in the delivery of preventive health services to a general medical clinic population. The patients were identified prospectively; data were obtained retrospectively with review of the charts. The study was conducted in a primary care clinic staffed primarily by internal medicine residents in an urban academic medical center. Patients who were receiving continuity care in the clinic and who were scheduled for an appointment during the 4-week study period were eligible for inclusion. Patients were identified by the appointment schedule. Charts were reviewed for the delivery of preventive health services. Data were abstracted utilizing a standard instrument. We found that the rate at which services were provided varied considerably by service and over time. The techniques used provided some insight into methods for the evaluation of the delivery of preventive services. It should be possible to assess a clinic's performance over a range of services over its entire population over time. An understanding of this more global performance may provide a better tool for managers and researchers addressing these issues. There may be legitimate reasons for services not being provided. These issues are complex and require sensitive, detailed investigation. PMID- 8704502 TI - Outpatient management of diabetes mellitus in five Arizona Medicare managed care plans. AB - We report findings on the outpatient management of diabetes mellitus in Medicare beneficiaries enrolled in five Arizona Medicare-managed care plans. These findings are the baseline of an ongoing collaboration between the Health Services Advisory Group, Inc., Arizona's Peer Review Organization (PRO), and the five plans whose object is improved care of diabetes patients. The purpose of the study was to determine congruity between quality indicators identified by the five plans and the care actually received by diabetes patients enrolled in the five plans. The five plans agreed on a common set of quality indicators, including 10 services and 10 measures of patient status. Each plan has identified its diabetic population, 75 of whom are randomly selected each quarter by the PRO for chart review and inclusion in the study. The findings in this report cover two quarters of data. Data from chart review were examined to determine the extent to which actual practice reflected the indicators. The mean patient age was 71.8, and for most patients onset occurred between 55 and 69 years of age. About 25% had a positive family history, and we estimate the annual incidence of diabetes in this population to be about 1.1%. Mean hemoglobin A1c (HbA1c) was 8.9 +/- 2.1%; 46% were hypertensive; 42% continued to smoke cigarettes; 36% had retinopathy; 20% had proteinuria; and only 22% were on some kind of exercise program. Thirty-two percent were hospitalized during the 1-year baseline period, and the average number of outpatient visits per patient was 11.1 +/- 7.4. When care provided to diabetes patients enrolled in the plans was compared with the 10 quality standards identified by the plans themselves, only two of these standards was attained in more than 60% of patients: blood pressure, 98.7%; and foot examination, 62.7%. Two standards were achieved less than one-third of the time: urine dipstick, 10.4%, and appropriate use of angiotensin-converting enzyme (ACE) inhibitors, 31.25%. The others were all between 40 and 55%. Of the 10 service standards, about one-third received 1-4, one-third received 5-6, and one-third received 7-10. Only 5% of patients received 9 or 10 services. Outpatient management of diabetes patients in managed-care plans is similar to that in fee for-service. When compared with fee-for-service or another HMO, a higher proportion of Arizona-managed care patients had HbA1c, and a much lower proportion had a dipstick test for urine protein. Values for other variables were usually within 10 percentage points of each other. Regardless of payment scheme, diabetes care is characterized by inconsistencies, omissions, and a lower than desirable level of services. Although few patients received most of the indicator services, diabetes patients are nevertheless high utilizers of medical care, both in and out of the hospital. The hospitalization rate is twice that of Arizona Medicare beneficiaries as a whole, and the number of office visits is three or four times that reported in other studies. Further, it seems that many visits are required to achieve even these modest service levels. Had the average number of visits been six or less, HbA1c rates, for example, would have fallen to less than one-third in three of the five plans. We believe that these data are conservative because it is likely that some and perhaps most of these indicators are underreported. It should be emphasized that these are baseline data whose purpose is to provide a basis against which subsequent improvements many be measured. PMID- 8704503 TI - Quality improvement and substance abuse: rethinking impaired provider policies. AB - Substance abuse is known to be our nation's number one public health problem. Physicians and other health providers can develop chemical dependency problems that create significant quality assurance and risk management dilemmas. Not all of society understands drug dependence to be a treatable medical disease and behavioral problem. Sometimes impaired providers are ignored or punished rather than treated and rehabilitated. This results in an enormous waste of human and monetary resources. In the last 10 years, impaired physician programs have developed focus and credibility. But certain difficulties exist in these programs: namely, that such programs are affected by tensions between medical societies and state licensing boards; that substance-dependent providers escape detection by moving to other states; that existing impaired provider programs have limited practical application within the federal health care system; and that liability risks are incurred if institutions rely on impaired provider policies that do not include all members of the medical staff. Hence, we argue that existing impaired provider policies might be worth rethinking. To promote that dialogue, we offer a sample policy for consideration and review. It includes specific actions and procedures for the identification, referral, and reentry of impaired providers and allows for National Practitioner Data Bank reporting in that process. The Data Bank has been in place for only a few years and offers society and the health community a new opportunity to better control chemically dependent, licensed medical staff without dismantling existing impaired provider programs. The policies for managing chemically dependent health professionals are changing from a focus on stigma and prosecution to one of early identification, rehabilitation, and reentry. We hope to advance that process. PMID- 8704505 TI - Interview with: Nancy Spencer. Interview by Ina May Gaskin. PMID- 8704504 TI - Re-imagining birth. PMID- 8704506 TI - What it took to keep the midwives from prevailing in a fair marketplace. PMID- 8704508 TI - Listening to babies. PMID- 8704507 TI - National certification for professional direct entry midwives. PMID- 8704509 TI - Postpartum depression: another view. PMID- 8704510 TI - Ask the midwife--on HIV/AIDS and nursing mothers. PMID- 8704511 TI - NARM discontinues the registry examination. PMID- 8704512 TI - Interview with: Marsden Wagner, author of Pursuing the Birth Machine. Interview by Ina May Gaskin. PMID- 8704513 TI - Midwifery, hiv and aids. PMID- 8704515 TI - Midwives dilemma: damned if you do, damned if you don't. PMID- 8704514 TI - The United States has had the highest rate of teen pregnancies in the industrialized world. PMID- 8704516 TI - One birth affects another. PMID- 8704517 TI - Fissured nipples: a breast feeding complication. PMID- 8704518 TI - Midwifery in Denmark. PMID- 8704519 TI - 40 something--over the hill or just beginning the climb? PMID- 8704520 TI - On postpartum depression from the land downunder. PMID- 8704521 TI - Interview with Sister Angela Murdaugh. Interview by Ina May Gaskin. PMID- 8704522 TI - St John's wort as an antidepressant. PMID- 8704523 TI - Access to civil justice: Lord Woolf's visionary new landscape. PMID- 8704524 TI - Managing medical mishaps. PMID- 8704525 TI - Motor neurone disease. PMID- 8704526 TI - Methadone treatment for opiate addicts. PMID- 8704527 TI - Breaking Olympic records. PMID- 8704528 TI - Overhaul of medical negligence litigation announced. PMID- 8704529 TI - Technique for treating infertility may be risky. PMID- 8704530 TI - Female circumcision is curbed in Egypt. PMID- 8704531 TI - Russia is on brink of AIDS epidemic. PMID- 8704532 TI - St John's wort for depression--an overview and meta-analysis of randomised clinical trials. AB - OBJECTIVE: To investigate if extracts of Hypericum perforatum (St John's wort) are more effective than placebo in the treatment of depression, are as effective as standard antidepressive treatment, and have fewer side effects than standard antidepressant drugs. DESIGN: Systematic review and meta-analysis of trials revealed by searches. TRIALS: 23 randomised trials including a total of 1757 outpatients with mainly mild or moderately severe depressive disorders: 15 (14 testing single preparations and one a combination with other plant extracts) were placebo controlled, and eight (six testing single preparations and two combinations) compared hypericum with another drug treatment. MAIN OUTCOME MEASURES: A pooled estimate of the responder rate ratio (responder rate in treatment group/responder rate in control group), and numbers of patients reporting and dropping out for side effects. RESULTS: Hypericum extracts were significantly superior to placebo (ratio = 2.67; 95% confidence interval 1.78 to 4.01) and similarly effective as standard antidepressants (single preparations 1.10; 0.93 to 1.31, combinations 1.52; 0.78 to 2.94). There were two (0.8%) drop outs for side effects with hypericum and seven (3.0%) with standard antidepressant drugs. Side effects occurred in 50 (19.8%) patients on hypericum and 84 (52.8%) patients on standard antidepressants. CONCLUSION: There is evidence that extracts of hypericum are more effective than placebo for the treatment of mild to moderately severe depressive disorders. Further studies comparing extracts with standard antidepressants in well defined groups of patients and comparing different extracts and doses are needed. PMID- 8704533 TI - Double blind randomised controlled trial of effect of metoprolol on myocardial ischaemia during endoscopic cholangiopancreatography. AB - OBJECTIVE: To evaluate the effect of metoprolol, a beta adrenergic blocking drug, on the occurrence of myocardial ischaemia during endoscopic cholangiopancreatography. DESIGN: Double blind, randomised, controlled trial. SETTING: University Hospital. SUBJECTS: 38 (two groups of 19) patients scheduled for endoscopic cholangiopancreatography. INTERVENTIONS: Metoprolol 100 mg or placebo as premedication two hours before endoscopy. MAIN OUTCOME MEASURES: Heart rate, arterial oxygen saturation by continuous pulse oximetry, ST segment changes during endoscopic cholangiopancreatography (an ST segment deviation > 1 mV was defined as myocardial ischaemia), electrocardiogram monitored continuously with a Holter tape recorder. RESULTS: All patients had increased heart rate during endoscopy compared with rate before endoscopy, but heart rate during endoscopy was significantly lower in the metoprolol group compared with the placebo group (P = 0.0002). Twenty one patients (16 placebo, 5 metoprolol; P = 0.0008) developed tachycardia (heart rate > 100/min) during the procedure, and 11 patients (10 placebo, 1 metoprolol; P = 0.003) developed myocardial ischaemia. One patient in the placebo group had an acute inferolateral myocardial infarction. In the 10 other patients with signs of myocardial ischaemia during endoscopy the ST deviation disappeared when the endoscope was retracted. In all patients myocardial ischaemia was related to increases in heart rate, and 10 of the 11 patients had tachycardia coherent with myocardial ischaemia. CONCLUSIONS: Metoprolol prevented myocardial ischaemia during endoscopic cholangiopancreatography, probably through lowering the heart rate. Thus, tachycardia seems to be a key pathogenic factor in the development of myocardial ischaemia during endoscopy. PMID- 8704534 TI - Prospective study of the role of cardiac troponin T in patients admitted with unstable angina. AB - OBJECTIVE: To examine the prognostic significance and role in risk stratification of the biochemical marker troponin T in patients admitted with unstable angina. DESIGN: Single centre, blinded, prospective study of patients admitted with chest pain. SETTING: Coronary care unit of a district general hospital. SUBJECTS: 460 patients admitted with chest pain and followed up for a median of three years. 183 patients had a final diagnosis of unstable angina. MAIN OUTCOME MEASURES: Cardiac death, need for coronary revascularisation, or readmission with non-fatal myocardial infarction as first events. RESULTS: 62 (34%) unstable angina patients were troponin T positive. This group had significantly increased incidence rates of subsequent cardiac death (12 cases (19%) v 14 (12%)), coronary revascularisation (22 (35%) v 26 (21%)), death or revascularisation (33 (53%) v 40 (33%)), and death or non-fatal myocardial infarction (18 (29%) v 21 (17%)) compared with the troponin T negative group. In multiple logistic regression troponin T status was a highly significant predictor for the end points coronary revascularisation and cardiac death or revascularisation as first events. CONCLUSION: Troponin T in the serum of patients with unstable angina identifies a subgroup at higher risk of subsequent cardiac events and its measurement aids in risk factor stratification. The increased risk extends to two years after admission. Prospective randomised trials are required to identify optimum therapeutic strategies for this subgroup. PMID- 8704535 TI - Characteristics of fatal methadone overdose in Manchester, 1985-94. PMID- 8704536 TI - Patients' awareness of adverse relation between Crohn's disease and their smoking: questionnaire survey. PMID- 8704537 TI - Road traffic noise and psychiatric disorder: prospective findings from the Caerphilly Study. PMID- 8704538 TI - Retrospective study of influence of deprivation on uptake of cardiac rehabilitation. PMID- 8704539 TI - Schistosomiasis in travellers returning from sub-Saharan Africa. PMID- 8704540 TI - Prescribing injectable and oral methadone to opiate addicts: results from the 1995 national postal survey of community pharmacies in England and Wales. AB - OBJECTIVE: To establish the extent of prescribing injectable and oral methadone to opiate addicts and the practice characteristics and dispensing arrangements attached to these prescriptions. DESIGN: National survey of 25% random sample of community (high street) pharmacies through postal questionnaire, with four mailings. SETTING: England and Wales. SUBJECTS: 1 in 4 sample of all 10,616 community pharmacies, stratified by family health services authority. MAIN OUTCOME MEASURES: Data were collected on each prescription for controlled drugs currently being dispensed by pharmacies to misusers, describing the drug, form, dose, source (general practice or hospital; and NHS or private), and numbers of dispensing pick ups a week. RESULTS: Methadone was the opiate most commonly dispensed to misusers (96.0% of 3846 opiate prescriptions). 79.6% of methadone prescriptions were for the oral liquid form, 11.0% for tablet, and 9.3% for injectable ampoules. More than one third of all methadone prescriptions were for weekly or fortnightly pick up, with a further third being for daily pick up. Tablets and ampoules were even less likely to be dispensed on a daily basis. Private prescriptions were significantly more likely than NHS ones to be for tablets or ampoules, to be for substantially higher daily doses, and to be collected on a weekly or fortnightly basis. CONCLUSIONS: The distinctively British practice of prescribing injectable methadone was found to be widespread and, contrary to guidance, to be as prevalent in non-specialist as specialist settings. In view of the frequent crushing and injecting of methadone tablets, clearer more authoritative guidance is needed on the contexts in which injectable methadone (tablets as well as ampoules) should be prescribed and on the responsibilities for monitoring and supervision which should be attached. PMID- 8704541 TI - Role of community pharmacies in relation to HIV prevention and drug misuse: findings from the 1995 national survey in England and Wales. AB - OBJECTIVES: To establish activity levels of community (high street) pharmacies in the provision of HIV prevention services to drug misusers and to compare these findings with the levels identified in 1988. DESIGN: Self completion questionnaire (four mailings) to a random 1 in 4 sample of all community pharmacies, stratified by family health services authority. SETTING: England and Wales. SUBJECTS: Data provided by pharmacist in charge of the dispensary, on service provision at the pharmacy. MAIN OUTCOME MEASURES: Quantitative reports of current activity levels for (a) dispensing of controlled drugs to drug misusers, (b) sale of needles and syringes, (c) needle and syringe exchange. RESULTS: 74.8% response rate (1984/2654). In 1995, 50.1% (992/1980) of pharmacies were dispensing controlled drugs (mostly methadone), compared with 23.0% (562/2457) in 1988; 34.5% (677/1962) of pharmacies were selling injecting equipment, compared with 28.0% (676/2434) in 1988; 18.9% (366/1937) were providing a needle exchange service, compared with 3.0% (65/2415) in 1988. CONCLUSION: Activity levels increased substantially across all three service areas. Increased activity included greater individual activity as well as higher proportions of pharmacies participating. The network of community pharmacies represents an underused point of contact for this Health of the Nation target population. PMID- 8704542 TI - Guidelines for authors and peer reviewers of economic submissions to the BMJ. The BMJ Economic Evaluation Working Party. PMID- 8704543 TI - Potentially lethal bacterial infection associated with varicella zoster virus. PMID- 8704544 TI - Should insurance companies have access to patients' medical records? Access should be denied. PMID- 8704545 TI - Should insurance companies have access to patients' medical records? Denying access would do more harm than good. PMID- 8704546 TI - Caring for older people. Money problems and financial help. AB - Older people make up the largest low income group in the United Kingdom. This article gives an overview of financial help available for older people, including general information, such as pensions and low income benefits, and also particular support for those with disabilities, such as attendance allowance and assistance with residential or nursing home fees. Advice on an individual's entitlements is available from the Benefits Agency and, particularly for those with disabilities, from social workers. PMID- 8704547 TI - ABC of work related disorders. Occupational asthma and other respiratory diseases. PMID- 8704548 TI - Intensive care of severely head injured patients. Answers may have reflected perceived rather than actual management. PMID- 8704549 TI - Intensive care of severely head injured patients. National research is needed. PMID- 8704550 TI - Intensive care of severely head injured patients. Working party is looking at guidelines. PMID- 8704551 TI - Assay used in serum screening for Down's syndrome may depend on ethnic make up of population. PMID- 8704552 TI - Intensive care of severely head injured patients. Guidelines should be based on systematic reviews of the evidence. PMID- 8704553 TI - Why interferon beta 1b was licensed is a mystery. PMID- 8704554 TI - Non-Hodgkin's lymphoma and solar ultraviolet radiation. Data are inconsistent. PMID- 8704555 TI - Non-Hodgkins lymphoma and solar ultraviolet radiation. Basal cell carcinoma may be linked to haematological malignancy. PMID- 8704556 TI - Peanut and nut allergy. Baby massage oils could be a hazard. PMID- 8704557 TI - Peanut and nut allergy. Creams and ointments containing peanut oil may lead to sensitisation. PMID- 8704558 TI - Peanut and nut allergy. Serious adverse reactions to adrenaline are becoming more likely. PMID- 8704559 TI - Peanut and nut allergy. Study was not designed to measure prevalence. PMID- 8704560 TI - Peanut and nut allergy. Reduced exposure might increase allergic sensitisation. PMID- 8704561 TI - Peanut and nut allergy. Sesame allergy is also a problem. PMID- 8704562 TI - Value of ECGs in identifying heart failure due to left ventricular systolic dysfunction. PMID- 8704563 TI - Inequality in income and mortality in US. Lower mortality in Hispanic population may have affected findings. PMID- 8704564 TI - Monitoring may need to be prolonged in patients given warfarin and amiodarone. PMID- 8704565 TI - Continuing rise in emergency admissions. GPs' low morale is contributing factor. PMID- 8704566 TI - Continuing rise in emergency admissions. Reduce time from referral by GP to first outpatient attendance. PMID- 8704567 TI - Continuing rise in emergency admissions. Visiting elderly patients at home immediately after discharge reduces emergency readmissions. PMID- 8704568 TI - Methods used in economic evaluations of prenatal screening are disputed. PMID- 8704569 TI - More paediatric renal donors should be used for adult recipients. PMID- 8704570 TI - Rights and responsibilities in HIV prevention. Article was misleading. PMID- 8704571 TI - Rights and responsibilities in HIV prevention. AIDS might have been tackled differently if it had presented as a disease of women. PMID- 8704572 TI - Maternal mortality in India. Maternal mortality is falling in India but at slower rate than in Britain. PMID- 8704573 TI - Maternal mortality in India. Preventable ill health occurs in Britain too. PMID- 8704574 TI - Rights and responsibilities in HIV prevention. HIV negative people should be responsible for protecting themselves. PMID- 8704575 TI - BMJ's embargo should not be broken. PMID- 8704576 TI - GMSC's advice on intrapartum care. Committee is out of touch with needs of profession. PMID- 8704578 TI - Maternal mortality in India. Training for overseas doctors may not be appropriate. PMID- 8704577 TI - GMSC's advice on intrapartum care. GMSC's advice differs from royal college's. PMID- 8704579 TI - [The genetic characteristics of the analgesic action of substance P, bradykinin and thyroliberin in the rats of 2 inbred strains]. AB - A participation of nonopiate peptide brain systems in the mechanism of individual pain sensation was studied. The experiments were carried out on rats of two inbred strains, Fischer-344 and WAG/G, which exhibit different threshold of thermal pain sensation. The action of nonopiate oligopeptides (substance P, bradykinin, and thiroliberin) on the latent period of tail immersion from hot water was studied. It was found that significant analgesic action of substance P, bradykinin, and thiroliberin is more pronounced in rats more sensitive to thermal stimuli (WAG/G). The rats with reduced thermal sensitivity (Fischer-344) are shown to be insensitive to analgesic action of substance P, bradykinin, and thiroliberin. PMID- 8704580 TI - [Cerebral blood flow and thrombocyte aggregation at different times of hypokinesia and their regulation by antiaggregants]. AB - Changes in local cerebral blood flow and in microcirculatory channels in the cortex of rat brain were studied in conditions of early and remote hypokinesia. The rheological parameters of blood were studied in patients which stayed in ber for a long time in traumatological hospitals. We showed that by the 60th day of hypokinesia and in conditions of active readaptation the disordered cerebral blood flow starts to stabilize and tendency to normalization of some parameters of the cerebral blood flow and microcirculatory channels become clearly seen. The study of the functional state of the platelets showed that prescription of antiaggregants along with physical training can prevent thromboembolic aggravations associated with hypokinesia. PMID- 8704581 TI - [The effect of lithium derivatives of GABA on blood circulation and metabolic indices in the brain under ischemia]. AB - In acute experiments on anesthetized cats with cerebral ischemia in conditions of autohemoperfusion of cerebral and peripheral vessels with a stable volume of blood we found that lithium oxybutirate and new GABA derivatives: LOS 1-84 and LOS 5-79 affect the cerebral blood flow and metabolism in the brain. Prophylactic intravenous injection of lithium oxybutirate did not affect the development of postischemic hyperperfusion but inhibited postischemic hyperperfusion. The compounds prevent the development of postischemic phenomena and promote the recovery of metabolism in the brain. PMID- 8704582 TI - [The effect of tyrosine-containing dipeptide on neuroglycopenia-induced changes in the activities of the sympathetic nervous system and of the systemic hemodynamics in waking rats]. AB - A modulatory role of the precursor of catecholamines--tyrosine (TYR)--in 2 deoxyglucose (2DG)-induced changes in blood pressure (BP), renal sympathetic nerve activity (renSNA), plasma levels of epinephrine (EPI) and norepinephrine (NE), total spillover of NE (TSO), and sensitivity of baroreceptor reflex (SBR) is studied. Chronically instrumented male Wistar rats were injected with 2DG (125 - 500 mg/kg iv), and BP, heart rate (HR), renSNA, TSO, EPI, NE, plasma clearance NE (CINE) and SBR were monitored before and after 2DG administration. The responses to 2 DG in control rats were compared with those in rats pretreated with tyrosile-tyrosine dipeptide (TYR-TYR). SBR was estimated by bolus i.v. sodium nitroprusside injection. 2DG evoked an increase in renSNA by 23%, 40-fold and 2.5-fold increases in EPI and NE concentrations, 2,7-fold increase in TSO, a decrease in HR by 5% in 45 min without reliable changes in BP and CINE. TYR-TYR administration did not affect 2DG-induced changes in HR, TSO, EPI and NE concentrations, but potentiated changes in renSNA by 53% and significantly increased BP by 10%. 2DG produced an increase in the chronotropic (by 87%) and sympathetic (by 72%) components of SBR. TYR-TYR restored the normal values of SBR in 2DG-treated rats. Thus, tyrosine modulates 2DG-induced changes in hemodynamic parameters and renSNA and normalizes SBR via central mechanisms. PMID- 8704583 TI - [The clinical significance of selenium deficiency in patients from the Samara region with cardiovascular diseases and its correction with the preparation Selena]. AB - Selenium concentration in blood, glutathionperoxidase (GSH-Px) activity in erythrocytes and lipid peroxides in plasma were studied in patients with ischemic heart disease, hypertrophic (HCMP) and dilated cardiomyopathy (DCMP). Patients with IHD, HCMP and DCMP revealed depressed activity of GSH-Px with activation of lipid peroxides associated with lowered selenium content in blood. The selenium content in blood was lower in patients with severe forms of IHD Daily intake of 300 micrograms selenium with "Selena" during a month increases selenium concentration by 71% in patients with IHD and DCMP. This increase change inversely with primary selenium concentration in plasma. Plasma Malone dialdehyde concentration decreased by 17%. The results obtained suggest participation of selenium in cardiomyopathy and IHD pathogenesis, thus forming a basis for selecting the patients with selenium deficiency for its further correction with "Selena." PMID- 8704584 TI - [The action of diuretics in renal ischemia]. AB - A protective action of lasix, dichlothiazide, and triampur (dichlothiazide + triamterene) was studied in experiments on rats. Ischemia was simulated by obstruction of kidney vessels and ureter for 90 min. Lasix and dichlothiazide produced a protective effect in renal ischemia and at the same time resulted decrease of lifetime of experimental rats. Triampur increased the lifetime and decreased the losses of potassium in kidney tissue. PMID- 8704585 TI - [The influence of the GABA-B receptor antagonists 2-hydoxysaclofen and faclofen on the sedative effect of fenibut and baclofen]. AB - Pretreatment of NIH-Swiss mice with HYS (8 mg/kg) is shown to reduce the inhibiting effect (namely, the reduction of the locomotion rate and the number of horizontal movements) of PHE (80 mg/kg) and do not affect the action of an equipotent dose (4 mg/kg) of BAC. BAC induced shortening of the locomotion period is reduced by HYS. HYS appeared to be inactive with respect to inhibiting effect of PHE. PHE and BAC shortening of the distance covered was insensitive to HYS administration. HYS reduces decrease in the rate and duration of rearings induced by PHE. PHA (8 mg/kg) do not change the sedative effect of PHE and BAC on locomotion, attenuated the PHE-induced inhibition of rearings and increase the effect of BAC. It is suggested that GABA-B receptors affect the inhibitory action of PHE rather strong than that of BAC and differ in the inhibition of horizontal and vertical components of the locomotor activity of mice. PMID- 8704586 TI - [The action of adrenergic agonists and antagonists on kidney function and arterial pressure depending on the mineral component of the diet]. AB - The experiments on rats showed that high consumption of sodium chloride promotes the diuretic action of phenylephrine (mesaton), the alpha-adrenergic agonist, and prolong its pressor effect. In supplementation of diet with sanasol and hyposol, a new substitute of common salt, phenylephrine, causes the renal effects typical of beta-adrenoceptor stimulation. Both the sanasol and especially hyposol decrease the strength and duration of phenylephrine pressor action and increase the duration of isadrine depressor effect. Thus, the reactivity of alpha- and beta-adrenergic mechanisms depend on the mineral composition of diet, which is able to change the pharmacodynamics of cardiovascular agents. PMID- 8704587 TI - [The protective action of limontar in pregnant animals with acute hypoxic hypoxia]. AB - A comparative study of individual resistance of rats to acute hypoxic hypoxia was performed. It was shown that individual resistance of the organism to hypoxia significantly decreases in the course of pregnancy. Limontar is shown to be active antihypoxant. Limontar administered during the last third of pregnancy stimulates protective-adaptive reactions in the females, thus increasing their resistance to hypoxia. PMID- 8704588 TI - [The effect of new synthetic prostanoids on the indices of the blood coagulation system]. AB - In the experiments on rabbits it was shown, that the administration of prostanoids IOS 3933 caused a reduction of ADP- and collagen induced platelet aggregation, platelet adhesion to collagen and elevation of t-PA level. This prostanoid prevented both a decrease in platelet count and reduction of AT III activity, but did not affect the protein C level during thromboplastin infusion. PMID- 8704589 TI - [The effect of cobalt chloride on hemoxygenase activity, on bilirubin metabolic homeostasis in the brain and on serum albumin transport function in animals]. AB - Administration of cobalt chloride (everyday dosage 50-100 mumol/kg) to rats in the course of pregnancy and first two days of postnatal period increases the activity of hemoxygenase + NADPH biliverdinreductase enzyme system in the brain of adult rats and induce this activity in new-born rats. Functional activity of serum albumin connected with bilirubin transport, the pH value in blood and bilirubin concentration in the brain of adult and new-born rats virtually do not change, whereas bilirubin concentration in the serum of pregnant rats received CoCl2 raises. When the same doses of cobalt chloride are administered to pregnant rats in combination with the mixture of high fatty acids and acetylsalicylic acid the activity of this enzyme system in the brain increases, functional activity of serum albumin connected with bilirubin transport decreases, bilirubin concentration in the brain raised (serum bilirubin concentration being comparatively low) at the same pH value in blood of both pregnant and new-born rats. PMID- 8704590 TI - [A comparative study of the anabolic action of ecdysten, leveton and Prime Plus, preparations of plant origin]. AB - A new class of anabolic substances includes different saponins, originated from Leuzea rhaponticum sp. The effect of these substances on the human body was studied by caliperometric measurements of body folds. All tested substances taken orally for three weeks diminished fat content under conditions of daily aerobic anaerobic training. Ekdisten and Prime Plus (combination of ekdisten and pure protein) elevated the muscle mass. The same effect of leventon was less pronounced. We showed that all substances studied elevated the magnitude of "total work", estimated per 1 kg of body weight. Ekdisten and Prime Plus again appeared to be most efficient. The prospects of using the saponins in sport medicine are discussed. PMID- 8704591 TI - [The immunomodulating action of polyunsaturated phospholipids]. AB - Intravenous injection of essential to Wistar rats stimulates the immune response to sheep's red cells and also induces the appearance of heavy red cells in animal's blood, thus causing the secretion of immunomodulating factor by sticking spleen's cells to glass. Physical training (swimming) suppresses the immune response to sheep's red cells. This effect is connected with light red cells. Essential injection to swimming rats normalizes the immune response and prevents the appearance of light red cells in blood of animals which possess the immunosuppressing activity. PMID- 8704592 TI - [The immunocorrective activity of myelopeptide-1 and myelopeptide-2 in cyclophosphane-treated mice]. PMID- 8704593 TI - [The toxicological characteristics of the interaction of cholinolytics with aminostigmine--a new reversible cholinesterase inhibitor]. AB - This investigation has been carried out on albino mice. In the first series of experiments, we determined the dependence of LD50 of aminostigmine on different doses of 10 cholinolytics, and conversely, the dependence of LD50 of cholinolytics on aminostigmine administration. The initial slopes of the dose effect curves were calculated. This data form the basis for evaluation of the character and degree of interactions. We established that benactyzine, spasmolytin, ftoracizin, arpenal, atropine, ganglerone, and methacin at low doses exhibit antagonism (with decreasing activity), whereas at high doses they exhibit synergism to the toxic effect of aminostigmine. In the interaction with aminostigmine, pirenzepine and amitriptyline (M1--cholinolytics) reveal a slightly pronounced antagonism, whereas aetyrophene, a selective central N cholinolytic, displays mutual synergism. In the second series of experiments we showed that a combination of M1-, M2-cholinolytic atropine with M1- cholinolytics does not change the efficacy of the prophylaxis of aminostigmine and physostigmine poisoning, whereas a combination with N-cholinolytic increases it. PMID- 8704594 TI - [The pharmacological selectivity of cholinesterase inhibitors is due to their antagonism with calcium ions]. AB - The inhibitory activity of O-carbamoyl acylhydroxymoylchlorides was studied on the purified specimens of acetylcholinesterase from human erythrocytes and butirylcholinesterase from the horse blood serum. The pharmacological activity of these compounds was studied on isolated organs of rats. It was shown that introduction of N-aryl substituent in carbamate molecule increases the anticholinesterase activity and efficacy of blocking the neuromuscular conductivity. However it significantly diminishes the spasmogenic effect of N arylcarbamates on the ileum. An increase in N-arylcarbamate concentration to 0.02 0.04 mM and higher results in a spasmolytic effect on the ileum which can be attributed to their "side" calcium blocking action. PMID- 8704595 TI - [Neuromediator systems in the mechanisms of remission and recurrence in alcoholism]. AB - Experiments on chronically alcoholic rats have revealed profound changes in the properties of the brain dopamine, serotonin, GABA and opiate systems 48 hours and 2 months of alcohol deprivation. With this, a single administration of ethanol showed its increased sensitivity of striatal and hippocampal receptors both during short- and long-term alcohol deprivation, the properties of these receptors tended towards recovery. It may be suggested that the above changes in the receptor apparatus of the neurotransmitter systems underlie the mechanism of remission and recurrence in alcoholism. PMID- 8704596 TI - [The efficacy of using synthetic carbon-mineral sorbents in combined radiation thermal lesions]. AB - The effect of enterosorbents of group "SUMS" on the manifestations of endogenous intoxication syndrome and dysfunction of small intestine was studied in rats with combined radiational-thermal injuries. The "SUMS" enterosorbents are shown to reduce the general toxemia, and promote the recovery of intestinal wall digestion enzyme activity and glucose absorption. PMID- 8704597 TI - [The mechanisms of the thermal protective action of atsefen]. AB - The study of thermoprotective properties of acephen (300 mg, single dosage) showed an increase (compared to that of placebo) in the maximum endurable time under thermal loading in healthy volunteers: optimization of evaporative heat exchange regime; reduction of the final values of reactive anxiety and fatigue and decrease in the rate of skin temperature growth. In combination with the data of biochemical analysis of peripheral blood which demonstrated positive changes in hormone, lipid, protein, and energy metabolism as well as in vegetative regulation, the results testify to a substantial thermoprotective potential of acephen attributed to the wide range of its pharmacological activity. PMID- 8704598 TI - [The formation and development of pharmacological research in Russian aviation and space medicine (on the 60th anniversary of the Institute of Aviation and Space Medicine of the Ministry of Defense of the Russian Federation)]. PMID- 8704599 TI - [Cardiointervalography in the practice of experimental and clinical pharmacology]. AB - The use of cardiointervalography (a method for taking into account the variability of cardiac rhythm) in solving the pharmacological problems both in the experiment and in clinical practice is considered. The method allows a rapid estimation of pharmacological sensitivity in human and animals. The method makes it possible to predict the drug tolerance development, and determine the period (day, month, and year) most suitable for drug administration. PMID- 8704600 TI - [Prodigiozan modulation of the specific activity of antidepressants]. AB - The experiments carried out on mongrel male mice showed that bacterial lipopolysaccharide Prodigiozan inhibit the activity of P-450-dependent monooxygenases of the liver, thus changing the activity of antidepressants- amitriptyline and imipraminum. The data on biological activity were tested in "tail suspension" test. Prodigiozan exhibited no central effect. Changes in biotransformation processes after single administration of both drugs resulted in the delay and prolongation of the effect. Prodigiozan prevents the induction of microsomal enzymes caused by antidepressants. Amitriptyline do not affect the immunostimulating action of prodigiozan, whereas imipraminum inhibited prodigiozan activity. PMID- 8704601 TI - [The correction with nooglutil and L-pyroglutamyl-D-alanine amide of cognitive disorders in rats due to intrauterine hypoxia]. AB - Novel nootropic compounds, nooglutyl (N-5-hydroxy(nicotinoyl)-L-glutamine acid, 25 mg/kg/day) and L-pyroglutamyl-D-alanine amide (1 mg/kg/day) administered intracutaneously from the 8th to 20th day of life prevent from movement hyperactivity in "open field", disturbances in ability to training and in memory in an alternate test and in tests of passive and active avoidance and normalize behavior of the adult mail rats (subjected to two-hour hypobaric hypoxia in on the 15-day of intrauterine life, vacuum corresponded to the height 8500 m) in extrapolation avoidance test. Additionally, nooglutyl recovered the normal growth of rats in the first month of their life, prevented from deceleration of investigating behavior of adults animals and disturbances of the mink reflex in them. PMID- 8704602 TI - [The effect of antidepressants on the catecholamine content of the rat hypothalamus and adrenals]. AB - Chronic administration of depressants of various classes in rats and a single administration of reserpine result in changes in catecholamine exchange in both hypothalamus and adrenal glands. Antidepressants prevent from a sharp decrease in level of hypothalamic dofamine, induced with reserpine. The importance of central dofaminergic mechanisms for specific effect of compounds is emphasized. PMID- 8704603 TI - [The effect of melatonin on rat behavior in a conflict situation]. AB - The effect of melathonine on behavior of rats under conditions of conflict situation is studied in experiments conducted in morning evening hours. The drug suppressed the response of rats on painful irritation and increased a number of taking water from drinking bowl. This effect of melathonine was more pronounced in the evening trials. The effects of melathonine were comparable with the influence of conventional benzdiazepine tranquilizer, diazepine, on the animal behavior. PMID- 8704604 TI - [The hypotensive effect of clonidine during direct and radiotelemetric recording of the arterial pressure in rats]. AB - A hypotensive effect of clonidine in non-narcotized intact and aorta baro denervated rats is studied under conditions of minimization of stress actions (radiotelemetry) and under standard conditions of direct recording arterial pressure (AP). Direct AP recording is shown to determine an increase in background AP in baro-denervated rats, but not in control rats. An increase in background AP level under conditions of direct recording is not accompanied with decreasing hypotensive effect of clonidine in baro-denervated rats. PMID- 8704605 TI - [The pharmacokinetics of different drug forms of nifedipine when used singly and in a course as monotherapy and in combination with Cordanum and triampur preparations in patients with arterial hypertension]. AB - Pharmacokinetics of three drugs derived from nifedipine: corinfar, corinfar retard, and SL adalate in the cases of a single and course administration in patients with arterial hypertension and the effect of cordanum and triampur on pharmacokinetics of corinfar retard in combined repeated administration have been studied. The studies were carried out in 6 groups of patients with arterial Hypertension, each group included 10 patients. Nifedipine concentration in blood plasma was determined using a special HPLC procedure within 24 h after administration of the drugs at a dose 20 mg. A pharmacokinetic characteristics of new drug adalate SL with two-step liberation of nifedipine. A possibility of autoinhibition was noted for corinfar and adalate SL in course therapy. A conclusion was made that cordanum and triampur did not affect the pharmacokinetics of corinfar retard. PMID- 8704606 TI - [Verapamil binding with the blood erythrocytes and cardiac output in myocardial infarct patients]. AB - In 14 patients with myocardial infarction the degree of verapamil binding with blood erythrocytes after a single oral administration (80 mg/kg) and the dependence pharmacological effect, the minute heart output (+ % MOC), on verapamil content in plasma and erythrocytes were studied. Intraindividual variability of verapamil distribution coefficients between erythrocytes and plasma (RBC/Ptot) is 9.7 +/- 2,3%, and it differs reliably from values of inter individual variability of this index (33.7 +/- 3.3%). PMID- 8704607 TI - [The clinical efficacy of taufon in the combined treatment of patients with chronic circulatory failure]. AB - Monotherapeutic treatment of patients with chronic cardiac insufficiency of I and II functional class and patients of III FC with Russian drug taufon against the background of traditional therapy leads to improvement of objective and subjective indications of the disease. Taufon does not significantly affect central hemodynamics, but normalizes contractile myocardial function and increases tolerance to physical load. Taufon potentiates therapeutic efficacy of some drugs that makes it possible to reduce their doses. Pharmacokinetics is given. The efficacy and the absence of any toxic effects allow the authors to recommend the drug for treatment of patients with chronic cardiovascular as an drug additional to basic background therapy. PMID- 8704609 TI - [The influence of retinol, tocopherol and cimetidine on the ulcerogenic effect of Orthofen, indomethacin and naproxen]. AB - The influence of retinol, alpha-tocopherol, and cimetidine on ulceration induced by ortophen, indomethacin, naproxen was studied on rats. It was shown that alfa tocopherol, retinol, and cimetidine reduce the damaging effect of these drugs on the gastroduodenal mucosa of rats thus normalizing the phospholipid content. The influence of alpha-tocopherol and retinol on the side-effects of ortophen was studied in children with rheumatoid arthritis. Ortophen was found to cause gastroduodenal side-effects in more than 30% of patients and decrease the levels of vitamin A and E in children. The use of retinol and alpha-tocopherol in cotherapy decreases the frequency and severity of side-effects and reduces the vitamin deficiency. PMID- 8704608 TI - [The characteristics of the anxiolytic action of taftsin and its analog TP-7 on behavior and serotonin metabolism in the brain of rats with chronic deprivation of serotoninergic system activity]. AB - The effect of taftcine (Thr-Lys-Pro-Arg) and its analog TP-7 (Thr-Lys-Pro-Arg-Pro Gly-Pro) on rat behavior and serotonin brain exchange was studied in 95 adult male Wistar rats neonatally administered with 5,7-dihydroxytriptamine. Intracutaneous administration of both peptides (300 micrograms/kg) was found to result in weakening perception of stress situations; an increase in stability of investigation behavior and normalization of serotonin level in the brain of rats with chronic deprivation of serotoninergic system activity. Anxiolytic and psychostimulant activities of TP-7 are more pronounced than those of tafcine. PMID- 8704610 TI - [Membrane-active preparations as correctors of the nephrotoxicity of gentamicin]. AB - At intracutaneous gentamicine injections in rats (dose 50 mg/kg) within 10 days the simultaneous application of anticalcium drugs makes nephrotoxic signs more pronounced; whereas antioxidants and essenciale make them less pronounced. Phospholipid complex, containing phosphatidylserin, sphingomyelin, and phosphatidylethanolamine does not exhibit a marked nephroprotective effect. The found character of changes in content of phosphoinozytols in urine indirectly confirms the role of disturbances of their metabolism in mechanism of aminoglycoside nephrotoxicity. PMID- 8704611 TI - [The effect of calcium antagonists, partusisten, oxytocin and prostenon on uterine contractile activity in pregnant rats]. AB - Experiments carried out in pregnant rats show that combination of nifedipine with verapamil or partusystene (in a half dose) depresses contractility in a more pronounced extent than of these drugs in the case of separate application. Calcium antagonist, cardizem does not depress contractility of myometrium. PMID- 8704612 TI - [The transplacental transport and effect of morphine administered in the prenatal period on its level in adult rats]. AB - The morphine concentration in the placenta, plasma of pregnant rats and their fetuses and in the plasma of prenatally morphinized rats was determined with HPLC method. It was found that 10 min after the intraperitoneal morphine administration at a dose of 20 mg/kg the morphine concentration in the fetus blood plasma is 3 times as large and in the placenta is 2 times as large as in the blood plasma of pregnant rats. The morphine level in prenatally morphinized rats was reliably higher than that in the control animals. PMID- 8704613 TI - [The effect of new synthetic prostanoids on the aggregation activity of human thrombocytes]. AB - We found that a number of new synthetic prostanoids caused a prominent reduction of human platelet aggregation in the in vitro experiments. Moreover, two of this agents, IOS 3933 and IOS 4732, gives storage stability. Hemoxygenase activity, metabolic homeostasis of bilirubin in brain and transport function of serum albumin in animals receiving cobalt chloride. PMID- 8704614 TI - [The relationship between cell membrane adrenoreceptor response and insulinemia in hypertension patients undergoing calcium antagonist treatment]. AB - Dynamics of correlations between the absolute values and individual changes in adrenoception of erythrocytes and insulinemia was studied in 17 patients with hypertension (II stage) prior to and after 2-month treatment by calcium antagonists. Clear correlation is observed between the sensitivity of alpha- and beta- adrenoceptors and reduction of insulinemia after treatment with calcium antagonists (r = -0.77, p < 0.05). The effect of heterologous sensitization of cell membranes on metabolism in patients suffered with hypertonia after their treatment with calcium antagonists are discussed. PMID- 8704615 TI - [The efficacy of the immunomodulating action of Essentiale and the modified animal erythrocytes]. AB - A relative order in reaction of immune system on antigenic irritant and its stability with respect to non-antigenic irritants is lowered to a greater extent in animals not able to swim, and in swimming animals it decreases to a smaller extent than in the case of administration of erythrocytes modified with essential (EME). After EME injections a phase of decreasing immunological reactivity with respect to T-dependent antigen; when essential is administered in animals the phase of immunosuppression is not observed. Essential normalized immune response to T-dependent antigen, if it is administered before physical load, combined with it, or after it. Injection of EME before the physical load increases its immunosuppressive effect. PMID- 8704616 TI - [The pharmacokinetics of propranolol and its conjugated metabolites in patients with chronic ischemic heart disease and arterial hypertension with a one-time and course intake of the drug]. AB - Pharmacokinetics of propranolol (P), 4-hydroxy-propranolol sulfate (4HOP-Sulf), and glucoronides of pharmacologically active S-enantiomer P (S-PG) and ballast R enantiomer of P (R-PG) in the blood serum of 21 patients with chronic ischemic heart disease and/or arterial hypertension has been studied at a single and course oral P administration. The values od AUC and T1/2 for potentially active 4HOP-Sulf were significantly higher than those for unchanged P at a single and course administration. The values od AUC and T1/2 for for S-PG were approximately three times higher than those for P-PG after both a single and course administration. Thus the results presented show that potentially active 4HOP-Sulf and S-PG (which undergoes a partial deconjugation in an organism at oral administration) may contribute essentially to the value and duration of the P pharmacological effect. PMID- 8704618 TI - [The acetylcysteine correction of disorders of the oxidative processes in a toxic liver lesion]. AB - The damage of the liver in albino rats induces by CCl4 was followed by the disorders in the free radical, microsomal, and mitochondrial oxidation. The administration of acetylcysteine to the animals in a dose of 500 mg/kg 3 times a day resulted in normalization of chemiluminescence parameters of the liver tissue, the rate of oxygen consumption by microsomes and mitochondria of the hepatocytes, and in normalization of the succinatdehydrogenase, cytochromooxidase, H-ATP-ase activity and ATP concentration. PMID- 8704617 TI - [The hepatoprotective and cholagogic action of glycyrrhizic acid derivatives]. AB - The experiments on rats with toxic hepatic damage induced by tetrachloromethane have shown that the derivatives of glycyrrhizinic acid (5NGA) promote a decrease in the rate of lipid peroxide oxygenation in the hepatic tissue homogenate and the blood serum, in the inhibition of organospecific enzyme activity (BHMT, ALT, AST) and increase in choleresis. The data obtained testify to the hepatoprotective activity of the derivatives of glycyrrhizinic acid. PMID- 8704619 TI - [A pharmacological analysis of the antiedematous action of GABA-ergic and opioidergic agents in craniocerebral trauma]. AB - Experiments on rats with skull inquiry show that biculline (dose 2 mg/kg) removes antiedematous effect of fenibut (50 mg/kg), sodium hydroxybutirate (200 mg/kg), promedol (1 mg/kg), and synthetic analogs of encephalines DAGO and DSLET (100 mg/kg). Naloxon at a dose 1 mg/kg blocks antiedematous effect of ligands of opiate receptors, but does not change the effect of fenibut and sodium hydroxybutirate. The presence of close inter-regulatory interactions between GAMK ergic and opioidergic systems in forming the traumatic correction is suggested. PMID- 8704620 TI - [The effect of haloperidol at a low dose on the development of experimental parkinsonian syndrome]. AB - Effect of galoperidole administrated in a small dose on changes in movement activity of mice in formation of hypokinetic syndrome in them using 1-methyl-4 phenyl-1,2,3,4-tetrahydropyridine (MPTP) was studied. Administration of galoperidol (0.01 mg/kg) increases the stability of structures of nigro-striate system with respect to injuring effect of MPTP; galoperidol administration in the same dose after formation of hypokinetic syndrome favors the more rapid recovering movement activity of animals. One of possible mechanisms resulting in the effect mentioned above may be an increase in a number of post-synaptic receptors, induced by galoperidol administration that, in its turn, may be attributed to activation of immediate response genes. PMID- 8704621 TI - [Changes in the brain neuraminidase activity under the action of tsiklozil and amizil]. AB - The effect of cyclozyl and amizyl on the activity of soluble and membrane-bound neuraminidase of the rat brain is studied in the in vivo and in vitro experiments. Within 60 min after subcutaneous injection of cyclozyl in a dose of 0.5 microgram/kg of body weight the activity of soluble neuraminidase decreased by 45.5% whereas the activity of membrane-bound neuraminidase remained the same. In the experiments in vitro the activity of both enzyme forms did not change when the content of cyclozyl and mixture of general gangliosides in the incubation media was 2.0, 20.0, and 200.0 micrograms. Amizyl (2 micrograms) in the presence of general gangliosides increased the neuraminidase activity whereas in the presence of disialoganglioside GD1b reduced the activity. Various lipids (phospholipids, phospholipids + cholesterol, cerebroside + sulfatides) surrounding the enzyme exerted different effect on the activity of both neuraminidase forms. Thus we suggest active interaction of the system "gangliosides-soluble and membrane-bound neuraminidase" with M-cholinolytics (cyclozyl and amizyl) and important role of this "substate-enzyme" system in M cholinoreceptor functioning. PMID- 8704622 TI - [The effect of the subchronic administration of calcium-channel blockers on CNS excitability]. AB - The effect of a single day administration of nifedipine (10 mg/kg), cinnarizine (25 mg/kg), verapamile (25 mg/kg), fendiline (10 mg/kg), dilthiazem (10 mg/kg), and diazepam (0.5 mg/kg) on the dynamics of changing a convulsive dose of convulsant was studied on a model of intermittent intracutaneous administration of Corazole (10 mg/kg until convulsions) during 5 days. Repeated intermittent Corazole administrations were found to increase the sensitivity of animals with respect to convulsive effect of the drug. All calcium channel blockers and diazepam increased the convulsive dose of Corazole. In accordance with the hypothesis of anxiety disturbances based on nonspecific CNS activation, a possibility of application of calcium channel blockers as potential anxiolytics is considered. PMID- 8704623 TI - [The effect of reserpine on epiphysial-adrenocortical relations]. AB - Reserpin at a dose inducing behavioral depression in rats, decreases the level of plasma corticosterone. Such effect is more pronounced in epiphysectomised animals and probably depends on drug induced central and peripheral catecholamines neurotransmission. PMID- 8704624 TI - [The role of adrenergic mechanisms in the anticonvulsive action of the new preparation acetylaminoazaleptin]. AB - In mice experiments the effects of adrenergic drugs (anaprilin, pirroxan, fetanol and alupent) on anticonvulsive activity of acetylaminoazaleptin (AAA) using the test of maximal electroshock and AAA effect on phenamin-induced manifestation (stereotypy and hyperthermia) were studied. It was demonstrated that anaprilin significantly (6-fold) increased anticonvulsive activity of AAA. The other drugs did not change AAA activity. In the experiments with phenamin AAA slightly weaken stereotypy and significantly reduced hyperthermia manifestation. PMID- 8704625 TI - [The use of the new cardioprotective agent thiotriazoline in the therapy of ischemic heart disease in older patients]. AB - The effect of tiotriazolin on the efficiency of antianginal therapy in aged patients with cardiac disease CD is studied. Clinical tests, functional (ECG, electrocardiotopography, cardiohemodynamics, etc.) and biochemical (indices of lipid peroxidation activity) methods of control were used as criteria of the treatment efficiency. Tiotriazolin was found to increase significantly the efficiency of basic drugs for CD treatment in aged patients. PMID- 8704626 TI - [The efficacy of hepatoprotective agents in experimental chronic hepatitis]. AB - Hepatoprotective drugs eplyr, essential, salsocollin, macsar and legalon in rats with chronic CCl4 hepatosis prevent fibrosis development, inflammatory infiltration of portal tracts, dystrophia and hepatocytes necrosis, reduce the content of hepatic glycoseaminoglycanes, oxyproline and different collagenic fractions in liver, weaken manifestations of cytolytic and cholestatic syndromes, improve antitoxic hepar function. PMID- 8704628 TI - [The excitatory effect of NMDA-receptor antagonists: a comparison of preparations and different forms of movements]. AB - Fourteen indices of spontaneous movement activity of mail NIH-Swiss mice were measured. All examined non-concurrent antagonists of NMDA receptors were found to stimulate locomotion and inhibit (or not change) the vertical activity component. Karbamazepin and diphenylgidantoin closed to them by structure and effect stimulated locomotion and rearings. Concurrent CPP antagonist and endogenous antagonist of receptor glycine sub-unit, kinurenic acid slightly inhibited locomotion. Phencyclidin and disocilpin induced stereotypy. Supposedly, increasing dopaminergic postsynaptic excitation forms the basis of stimulating effect. PMID- 8704627 TI - [The corrective properties of glycine in alcoholic intoxication in the fetal period of pregnancy]. AB - Natural metabolite glycine administered per os to rats in the fetus period of pregnancy in a dose of 1 mg/kg was found to exert a corrective action with respect to the toxic effect of ethanol in the mother-fetus system. Glycine prevents loss in the body weight, normalizes functional state of the nervous system and metabolic disorders in the maternal organism, improve the redox processes in the lymphocytes changed under alcoholization, and protect the fetus both on the metabolic and microstructural levels. PMID- 8704629 TI - [The effect of the joint administration of heparin and chitosan sulfate ether on hemostatic function]. AB - We studied anticoagulant effects of combined administration of heparin (H) and chitosan sulfate ether (CS) (specific activity 20 UE/mg) in the ratio 1 : 1. CS enhanced anticoagulant activity of heparin in rabbits by a factor of 1.95 +/- 0.15. The intravenous injection of the mixture in a dose of 0.5 mg(H)/kg + 0.5 mg(CS)/kg and heparin injection in a dose of 1mg/kg induced the same effect. Haemorrhagic effect of this mixture was less pronounced compared to heparin, anticoagulant and antithrombotic activities remained the same. The mixture was found to decrease a number of platelets, however, this was also less pronounced compared to heparin. Thus, the use of the mixture CS + H (1 : 1) instead of double heparin dose resulted in the same effect. PMID- 8704630 TI - [The effect of the quaternary ammonium salt of conidine oligomer-25 (QAS CO-25) on the activity of prothrombin complex factors]. AB - Quaternary ammonium salt of oligomer-25 conidine (QAS O-25C) was used for regulation of hypocoagulation effect of Syncumar--anticoagulant of indirect type of action. The experiments were carried out on male rats. Experimental methods included thromboelastography, determination of thromboplastin time (after Quick) and determination of activity of single coagulation factors. It was shown that injection of QAS O-25C reliably do not affect blood coagulation. The combined action of QAS O-25C and Syncumar resulted in a twofold increase in the activity of factors IX and X within 24 h after the injection and in complete recovery after 48 h. After Syncumar injection per os we observed deep-hypercoagulation which was maintained even within 48 after the injection. The effect of QAS O-25C modulating the Syncular anticoagulant activity is probably connected with intensification of protein synthesis in the liver and, specifically, with an increase in the yield of active precursors of the factors of prothrombin complex. PMID- 8704631 TI - [Modification to the functional status of the hemostatic system with the use of berberine sulfate]. AB - The effect of berberin sulfate (single and chronic administration during 30 days in doses of 0.14-80 mg/kg) on the functional state of hemostasis system was studied on gamma-irradiated (dose 5.0 Gy) and intact Wistar rats and mice (CBA x C57BL)F1. Administration of the drug increases the number of thrombocytes, decreases the activity of factor XIII and promote blood coagulation. PMID- 8704632 TI - [The pharmacological properties of a direct-action anticoagulant from the herb Nonea poulla (L.) D. C]. AB - Nondialyzed fraction of the ammonia extract from Siberian grass Nonea poulla is studied. A stable hypocoagulemy (without cumulative activity) was found in the experiments in vitro and in vivo carried out on rats (intravenous administration of the extract). The LD50 was 150 mg/kg at the effective dose of 10 mg/kg. Intraperitoneal and intramuscular administration during 20 days exerted neither hypocoagulemic nor toxic effect on the haemopoesis system and did not change the body weight of the animals. PMID- 8704634 TI - [The immunomodulating action of preparations possessing membrane-protective properties under intensive physical loading]. AB - We found that essential, tocopherol acetate, and lidocaine hydrochloride enhance the immune response to T-dependent antigen under active physical loading. Essential induces the appearance of immunostimulating properties in heavy erythrocytes whereas tocopherol acetate, and lidocaine hydrochloride increase the resistance of light erythrocytes to the effect of blood serum compounds (extracted from swimming animals) which induce the immunosuppressive properties in them. PMID- 8704633 TI - [The effect of paracetamol and its combinations with acetylsalicylic and ascorbic acids on lipid peroxidation processes in the rat liver]. AB - Paracetamolum (PC) after i.p. administration to rats at the therapeutic (100.0 mg/kg) and subtoxic (500.0 mg/kg) doses was shown to induce significant and dose dependent elevation of the level of malone dialdehyde (MDA) as well as the increase in a certain extent of chemiluminescence (CL) being injected at the higher dose. Acetylsalicylic acid (ASA) at the same doses being injected at the lower one, induced the tendency toward the reduction of MDA level, while at the higher dose the rise of MDA content and CL intensity compared to PC have been observed. PC combination with ASA at therapeutic doses range (50.0 + 50.0 mg/kg) significantly diminished MDA formation, while at subtoxic doses range (250.0 + 250.0 mg/kg) such effect was not detected, although subsequent tendency toward CL intensity reduction was observed. Ascorbinic acid being administered in combination with PC and ASA had no influence on MDA formation but significantly weakened CL intensity when subtoxic doses of indicated drugs were used. PMID- 8704635 TI - [The immunotropic properties of biologically active products obtained from Klamath weed (Hypericum perforatum L.)]. AB - We carried out a systematic study of immunotropic activity of John's wort on the level of integrated fractions including all basic active substances of this plant. Both types of substances capable of increasing and suppressing the immunity were found in John's wort. Polyphenol fraction exhibits the immunostimulating activity with respect to the system of mononuclear phagocyte system, cellular and humoral immunity, and is capable of recovering the immune response in conditions of high-zone tolerance. Lipophilic fraction of this plant exhibits immunosuppressing properties with respect to cellular and humoral immune response. Immunotropic activity of polyphenol and lipophilic fractions correlate with their low toxicity. PMID- 8704636 TI - [The ecdysteroid fraction of the above-ground portion of Serratula coronata L. in the spontaneous E-rosette formation reaction and the agar migration test in vitro]. AB - Reaction of E-rosette formation and agar migration test in vitro showed that small concentrations (7.5.10(-13) -7.5.10(-8) M) of phytosteroid ecdysterone (20 hydroxyecdysone) induce the activation of human lymphocytes. The effect of higher concentrations (7.5.10(-7) -7.5.10(-5) M) is less pronounced (two-phase effect). PMID- 8704637 TI - [The pharmacological properties of preparations from the aziridinyl triazine group]. AB - The results of experimental studies of three drugs from aziridinyltriazine group, dioxadet, trisadet, and furizil, are presented. These drugs exhibit a pronounced therapeutic effect against a broaden spectrum of tumors grafted in mice and rats. Toxic effect of the drugs is exhibited mainly in suppression of blood formation. Other side-effects are slightly pronounced. One of the drugs, dioxadet passed the I and II stages of clinical trials and is recommended for application in chemotherapy of malignant tumors. PMID- 8704638 TI - [The biochemical and rheological properties of the blood during the performance of angiography using Ultravist]. AB - It is shown that Ultravist makes it possible to receive clear visualization of vessel channel in the zone "of interest" without changing functional conditions of heart vascular system and biochemical blood parameters (level of erythrocytes, bilirubine, urine nitrogen, activity of aspartataminotransferase). In concentration 30 mg/ml in vitro and in vivo Ultravist decreases a viscosity limit not affecting other rheological properties of blood. A mechanism of the found Ultravist effect and prospects of its application in practice for children are considered. PMID- 8704639 TI - [The comparative clinico-experimental characteristics of aminostigmine and galanthamine used for treating poisonings by choline-blocking substances]. AB - The results of application of cholinesterase inhibitors, aminostigmin and galantamin, for treatment of acute poisoning with cyclodol, dimedrol, and solutan of moderately grave condition are presented. Aminostigmin was shown to exhibit the more pronounced stable and universal effect. The experiments in animals showed that aminostigmine affected peripheral and central M-cholinoreactive structures and conjugated with them more actively than galantamin. Aminostigmin, but not galantamin increases the rate of dopamine circulation and content of cyclic guanozinemonophosphate in frontal brain of rats, and this effect is exhibited even under the conditions of N-cholinoreceptor blockade with amizyl. PMID- 8704640 TI - [The antagonism of phosphorus-containing heterocyclics with the anticholinesterase agent Phosphacol (paraoxon)]. AB - Effects of 15 derivatives of 2-R-oxo-(tioxo-3,5,5-trimethyl-1,2-oxaphospholan-3 ol (OPh) on lethality and protective action on cholinesterase (ChE) of mice brain and blood serum as a result of phosphacolum (paraoxonum) poisoning were investigated. Oxo and tioxo derivatives of OPh with O-methyl and O-ethyl, but not O-propyl and O-butyl radicals at phosphoric atom prevented the lethality of mice poisoned with phosphacolum and preserved brain and blood ChE of inhibition. Ethylic and phenylic OPh derivatives also prevented the lethality of poisoned animals, although without the protection of brain and blood plasma ChE. Combined application of OPh O-ethylic derivative with atropinum and dipiroxinum significantly potentiated antidotal effect m mice poisoned by phosphacolum. PMID- 8704641 TI - [A method for assessing the analgesic action of drug preparations]. AB - A method for objective evaluation of analgetic activity of drugs is proposed. The method consists in measuring changes in skin galvanic potential response to swallowing prior to and after administration of the drug to the patients within 2 3 days after tonsillectomy. The pain in the throat is of swallowing-induced character and thus results in orientation response which can be detected via changes in skin potential. The more the intensity of the pain the greater the response. The method is used in studying the analgetic effect of benzofurocain. PMID- 8704642 TI - [New approaches to antitumor therapy: the use of preparations acting on processes regulated by epidermal and/or alpha-transforming growth factors]. PMID- 8704643 TI - [The neuromediator mechanism of the additive action of the delta sleep-inducing peptide in experimental audiogenic epilepsy caused by hypokinesia]. AB - The DSIP influence on the rat cortex, thalamic and hypothalamic adrenaline, noradrenaline, DOPA, dopamine and serotonin level under normal, hypokinetic stress condition (1 h in duration) and experimental audiogenic epilepsy was studied. It was found, that a single intraperitoneal DSIP injection (12 microns/100 g body weigh, 1 h before placing the animals into the stress condition) prevented spontaneous epileptic activity development due to creation of optimal ratio between monoamines in brain structures. PMID- 8704644 TI - Cyclin kinase inhibitors add a new dimension to cell cycle control. PMID- 8704645 TI - Head and neck non-Hodgkin's lymphomas. AB - 74 patients with non-Hodgkin's lymphoma (NHL) in the head and neck were studied and 21 out of 74 (28.4%) had a primary extranodal location. The most commonly affected lymph nodes were the lateral cervical (48%); the extranodal NHL occurred most frequently in Waldeyer's ring (19%) and orbit (5.4%). Overall 10-year survival was 55.2% with a median survival of 42 months, and survival was higher in extranodal (67.3%) than in nodal locations (51.9%). No statistically significant difference between the survival of high-grade and low-grade tumours was observed. There was, on the contrary, a correlation between stage of the disease and survival of the patients. Most cases were B-cell lymphomas (91.8%). Nodal NHL presented at diagnosis in an advanced stage in a higher percentage (71.2%) than extranodal (48%). High-grade tumours seemed to affect mainly young people. PMID- 8704646 TI - Role of alcohol and tobacco in the aetiology of head and neck cancer: a case control study in the Doubs region of France. AB - A case-control study conducted from 1986 to 1989 using the Doubs Cancer Registry included 299 cases of head and neck cancer and 645 controls from the general population. The results provide an indicator of the respective roles of alcohol and tobacco in all these cancers and on the tumour site. The people who smoked more than one packet of cigarettes a day have a risk that is 13 times higher than that of non-smokers and those who drink more than one and a half litres of wine per day have a risk that is 34 times higher of developing head and neck cancer. The combined exposure of alcohol and tobacco is characterised by a high risk and can be described by a multiplicative model without interaction. The age at onset (below 18 years of age) and the duration of smoking (over 35 years) are high risk factors. The risk decreases after stopping smoking, but only casual smokers (less than 7 cigarettes per day) can hope to have the same risk as non-smokers within a period of 15 years. Subjects smoking only non-filter cigarettes have a higher risk (OR = 1.98) than those who smoke filter cigarettes. The same applies to those who roll their own cigarettes (OR = 1.93) or inhale the smoke (OR = 1.51). PMID- 8704647 TI - Risk factors which predict persistent cancer in the abnormal larynx following definitive irradiation. AB - Laryngeal abnormalities following definitive irradiation for carcinoma of the larynx are common. The objective of this study was to identify risk factors for persistent cancer in such patients who were found to have abnormal larynges following definitive irradiation. A retrospective evaluation of 185 consecutive patients undergoing primary irradiation for a glottic or supraglottic laryngeal squamous carcinoma treated between 1976 and 1990 at the Affiliated Hospitals of the Medical College of Wisconsin was performed. From chart review, data concerning site, stage, intent of treatment, smoking history, treatment dose, fraction size, failure patterns, and outcome were obtained. In addition, worrisome signs and symptoms including ulceration, dysphasia, odynophagia, airway distress, aphonia, blood, pain, oedema, aspiration, and pneumonia were recorded. Univariate association with failure and a persistently abnormal laryngeal examination was assessed using the Mantel-Haenszel test. The odds ratio was used to estimate relative risk associated with dichotomous risk factors. Disease-free and overall survival were estimated using Kaplan-Meier methodology. The log rank test was used to compare survival as defined by the levels of various risk factors. Two-year disease-free survival was 83% (T1 = 93%, T2 = 72%, T3/T4 = 66%). Primary failure was associated with the presence of an abnormal examination (P = 0.001), tracheotomy (P = 0.001), symptom index (P = 0.002), aphonia (P = 0.003), advanced T stage (P = 0.03), and lower total dose (P = 0.03). Of 151 patients who survived 6 months disease-free with an intact larynx, an abnormal examination was seen in those with advanced T stage (P = 0.002), supraglottic primary (P = 0.003), symptom index (P = 0.008), eventual failure at the primary site (P = 0.008), continued smoking (P = 0.01), and higher total dose (P = 0.01). The symptom index (total signs and symptoms of airway distress, aphonia, ulceration, pain, oedema, dysphagia, blood production, aspiration, pneumonia, and odynophagia) was correlated with primary failure and continued smoking. Of 37 patients with continually normal examinations, only 1 (3%) failed at the primary site. Of 102 who survived 6 months but with an abnormal examination, 22 (22%) eventually developed a primary failure. Persistently abnormal larynges are common after radiation therapy, yet not all harbour cancer. Risk factors for persistent cancer include stage, airway, total dose, and symptom index. Patients whose larynges return to normal after radiation rarely fail at the primary site. PMID- 8704648 TI - Second cancers occurring after cancers of the mouth and pharynx: data from three population-based registries in Australia, Scotland and Slovenia. AB - Data over at least 20 years from three large population-based registries in Europe and Australasia have been used to assess the risk of second primary tumours occurring after a cancer of the mouth or pharynx. These patients have previously been shown in clinical series to be at a particularly high risk of subsequent tumours, while data from cancer registries have shown conflicting results on the magnitude of the risk. In this study, patients were found to have between a 2-fold (Scotland and New South Wales) and 4-fold (Slovenia) increase in risk of a subsequent tumour over that in the population, although the actual risk in each centre was similar (between 2.8 and 3.1 per 100 person years). The risk remained for 10 years after diagnosis of the original tumour and was primarily in the upper aerodigestive tract. The most elevated risks (approximately 10-fold) were for tumours in the oral cavity and oesophagus. These data provide higher estimates of risk than previously reported from European cancer registries for second primary tumours and emphasize the need for close follow-up of patients who may represent an appropriate population in which to assess possible new chemopreventive agents. PMID- 8704649 TI - Retrospective epidemiological study of Burkitt's lymphoma in Israel and diagnosis by a conservative incisional biopsy. AB - The clinical features of 74 patients with Burkitt's lymphoma diagnosed at the Hadassah Medical Center in Jerusalem, Israel, are described. The prompt diagnosis of a child with high grade malignant jaw involvement of non-Hodgkin's Burkitt's lymphoma is reported. The use of the open biopsy with special emphasis on the incisional biopsy is discussed in light of the incidental exfoliation of the two primary molar teeth and one permanent successor in this case. PMID- 8704650 TI - Simultaneous production of G- and M-CSF by an oral cancer cell line and the synergistic effects on associated leucocytosis. AB - We established a novel cell line, TSU, from a oral cancer patient with marked leucocytosis. The culture supernatant of TSU cells promoted granulocytic colony formation by mouse bone marrow cells, indicating that TSU produced granulocyte colony stimulating factor (G-CSF). The concentration of G-CSF was 2.45 micrograms/mg protein, measured by enzyme-linked immunosorbent assay (ELISA). The maximum number of colonies induced by TSU culture supernatant was more than that achieved with recombinant human G-CSF (rhG-CSF) and the size of the colonies induced by TSU supernatant was obviously larger than those achieved with rhG-CSF. The activity of TSU supernatant was completely inhibited by antihuman G-CSF and macrophage-colony stimulating factor (M-CSF) antibodies, but was only partially inhibited by antihuman G- or M-CSF antibody alone. These results indicate that not only G-CSF but also M-CSF, both of which could be produced by TSU cells, are involved in causing leucocytosis; the results suggest that the synergistic production of G- and M-CSF could play an important role in the leucocytosis associated with oral cancer. PMID- 8704651 TI - Infrequent MDM2 gene amplification and absence of gross WAF1 gene alterations in nasopharyngeal carcinoma. AB - We have investigated the possible involvement of MDM2 and WAF1 gene alterations in the development of nasopharyngeal carcinoma (NPC). MDM2 and WAF1 were analysed in 46 primary NPCs by Southern blot analysis. Forty-five tumours showed a normal EcoRI hybridisation pattern and hybridisation intensity with a human MDM2 cDNA probe. One tumour had more intense normal size MDM2 hybridising bands. Densitometric scanning revealed a 10-12-fold MDM2 gene amplification, as compared with human placenta DNA. All 46 tumours showed normal size WAF1 EcoRI bands that hybridised with normal intensity. This is the first demonstration of MDM2 gene amplification in NPC. Nonetheless, our analysis indicates that gross structural alterations of the MDM2 and WAF1 genes are infrequent events in the genesis of NPC. PMID- 8704652 TI - Squamous cell carcinoma of the lips in a northern Greek population. Evaluation of prognostic factors on 5-year survival rate--I. AB - The purpose of this study was to determine the clinical features of squamous cell carcinoma (SCC) of the lips, along with its prognostic factors, in order to extend and update the information related to lip cancer in northern Greece and to provide a basis for international comparison. Records of 1510 patients with SCC of the oral cavity presented at the Theagenion Anticancer Institute of Thessaloniki, Greece from 1979 and 1989 were reviewed. The most common site for oral squamous cell carcinoma (OSCC) was found to be the lips (59.4%) as compared to 40.5% of intra-oral SCC. Males were affected more frequently, presenting a ratio of 9.2:1. The peak age of incidence was found to be the 6th decade for men and the 8th for women. Rural residents and outdoor workers were affected more than urban residents (79.9% versus 28.1%). Most of the patients were diagnosed in early categories and early clinical stages of the disease. Almost all (98.5%) were classified into T1 and T2 categories, and 92.9% into stages I and II. A total of 7.59% of patients presented with clinically-positive lymph-node involvement. Most of them were classified as an advanced stage of the disease. Primary surgical excision was performed on 60.14%, radiotherapy on 35.14%, a combination of these on 2.47%, and chemotherapy alone or in combination with the above regimens in 2.22% of the cases. The outcome was adequate for surgery, radiotherapy, and the combination of the two, since 91.3, 74, and 90%, respectively, survived for more than 5 years. An overall 5-year survival rate of 83.3% was found. Our findings showed that the survival rate was significantly influenced by the main prognostic factors, such as the size of the tumour, the lymph-node involvement, the clinical stage of the disease and the histologic differentiation. SCC of the lips continues to be the most common site of oral cancer development amongst the Greek population. The aetiologic significance of actinic radiation for SCC of the lips is confirmed by our findings. The main prognostic factors proved to significantly influence the survival of our patients. Our results support the fundamental principal of head and neck cancer, i.e. that early detection of the primary tumour is the best prognostic factor for increasing survival rates. Therefore, public awareness concerning the disease, and better education for health care workers which will provide a thorough knowledge for the prognosis of oral cancer and the factors influencing it, is necessary. PMID- 8704653 TI - Self-reported satisfaction with life and physical health in long-term cancer survivors and a matched control group. AB - The response to two validated questions assessing self-reported satisfaction with life and physical health were compared between 204 long-term head and neck cancer survivors from a randomised radiotherapy trial and 766 matched controls. The control group, participants in a large population health survey in a county in the middle of Norway, was matched to the patients with regard to age, gender, marital status/family type and education. Compared with the controls, the patients reported significantly lower satisfaction with life and physical health. Sixty-four per cent of the patients were satisfied with life and 44% felt strong and healthy while the percentages of the controls were 82% and 52%, respectively. Clinical and sociodemographic variables were poor predictors of the patients' responses. Clinicians must be aware of the psychosocial morbidity in the growing population of cured cancer patients, and intervention studies are needed in patients at risk, in order to determine whether this morbidity can be prevented, reduced, or treated. PMID- 8704654 TI - Photodynamic therapy and lip vermilion dysplasia: a pilot study. PMID- 8704655 TI - Electrophoretic forms of chitinolytic and lysozyme activities in ruminal protozoa. AB - Homogenates from a mixed ruminal protozoal population and a ruminal protozoon Entodinium caudatum were analyzed for chitinolytic and lysozyme activities by sodium dodecyl sulfate polyacrylamide gel electrophoresis. For chitinase activity, up to eight bands in mixed protozoa and seven bands in E. caudatum were detected. Estimated molecular mass ranged from 70 to 110 kDa. These enzymes did not display lysozyme activity. N-Acetyl-beta-glucosaminidase activity was also detected in both samples with an estimated molecular mass of 37 kDa. Lysozyme activity in mixed protozoa was present in two major and three minor bands, where one major band displayed the same motility as chicken egg white (CEW) lysozyme, and the other had an approximate molecular mass of 17.5 kDa. The latter remained active even when denatured in the presence of dithiothreitol and renatured under anaerobic conditions. Entodinium caudatum presented one major band coincident with that of CEW lysozyme and a minor band at the 17.5-kDa point. This study showed that protozoal chitinase and lysozyme activities are originated from several enzymes and that none of these enzymes exhibited both activities. PMID- 8704656 TI - Quantitative determination of H2-utilizing acetogenic and sulfate-reducing bacteria and methanogenic archaea from digestive tract of different mammals. AB - Total number of bacteria, cellulolytic bacteria, and H2-utilizing microbial populations (methanogenic archaea, acetogenic and sulfate-reducing bacteria) were enumerated in fresh rumen samples from sheep, cattle, buffaloes, deer, llamas, and caecal samples from horses. Methanogens and sulfate reducers were found in all samples, whereas acetogenes were not detected in some samples of each animal. Archaea methanogens were the largest H2-utilizing populations in all animals, and a correlation was observed between the numbers of methanogens and those of cellulolytic microorganisms. Higher counts of acetogens were found in horses and llamas (1 x 10(4) and 4 x 10(4) cells ml-1 respectively). PMID- 8704657 TI - A novel secondary metabolite relative to the degradation of PR toxin by Penicillium roqueforti. AB - A secondary metabolite different from PR-imine and PR-amide was produced in the liquid (YESC) and solid (buckwheat) culture medium of Penicillium roqueforti. We isolated and purified the compound in pure and colorless crystalline form. On the basis of elemental analysis, mass, 1H and 13C nuclear magnetic resonance, infrared, and UV spectroscopy, the compound was identified as PR-acid (C17H20O7). The structures of PR-acid and PR toxin (C17H20O6) are closely related. Moreover, we discovered that PR-acid disappeared concurrently with the PR toxin in the culture medium. Thus, we postulate that PR toxin is degraded to PR-acid in the culture of P. roqueforti. PMID- 8704658 TI - Properties of beta-lactamase from Pseudomonas syringae. AB - Pseudomonas syringae isolate BR2R produces tabtoxin, a beta-lactam-containing antibiotic, and the causative agent of wildfire disease of green bean (Phaseolus vulgaris). beta-Lactamase production has been suggested as the mechanism that protects P. syringae from tabtoxin. We sought to determine whether the organism produces beta-lactamase and whether the enzyme plays a role in protection from this antibiotic. P. syringae and mutants defective in tabtoxin production and resistance produce beta-lactamase. Three distinct beta-lactamases with molecular weights of 41,000 were identified. The isoelectric points of the proteins were 6.1, 6.8, and 9.2. The enzymes preferentially hydrolyze cephalosporin. This investigation demonstrates that the organism produces multiple beta-lactamases and describes characteristics of the proteins. PMID- 8704659 TI - Characterization of an unusual fluoride-resistant Streptococcus mutans isolate. AB - A fluoride-resistant Streptococcus mutans isolate NCH105 was characterized and compared with wild-type strain UA130. The growth and lactic acid production of strain NCH105 were found to be unaffected by the presence of fluoride at the initial medium pH values of 6.5 and 6. In addition, NCH105 was found to be capable of lowering the pH of the medium to approximately 5.5, which is the critical level where enamel demineralization begins in vivo. Lactic acid production, glucose uptake at pH 6.5 and 6, as well as ATPase activity at pH 5 were found to be unaffected by fluoride. Finally, strain NCH105 is capable of binding as well as the wild-type bacterium to artificial tooth pellicles. These results are unusual when compared with previously isolated fluoride-resistant mutants and suggest that NCH105 may have the ability to colonize the tooth surface and initiate dental caries. PMID- 8704660 TI - Promotion of bacteriophage induction and recombination by the cloned Bordetella pertussis recA gene is copy-number dependent. AB - Favre and coworkers (Favre et al., Biochimie 73:235-244, 1991) previously reported that the Bordetella pertussis recA gene present at high copy number could promote a low frequency of recombination, but not bacteriophage induction in Escherichia coli RecA- mutants. Reexamination of these phenotypes demonstrated that, in contrast to the previous study, when this gene is present at high copy number, it can stimulate a 2- to 4-log frequency of bacteriophage induction in the presence of mitomycin C, but no appreciable spontaneous induction. The cloned gene, whether it was present in high or low copy number, also promoted a low frequency of intrachromosomal recombination of two duplicated genes in Escherichia coli. These results suggest that a high concentration of the B. pertussis RecA protein is required to promote high-frequency mitomycin C stimulated bacteriophage induction, but it facilitates intrachromosomal recombination at a very low frequency in E. coli RecA- mutants. The ability of the B. pertussis RecA protein to promote mitomycin C induction and its inability to appreciably stimulate spontaneous induction of bacteriophage suggest that this protein possesses a unique phenotype compared with other RecA proteins. PMID- 8704661 TI - Meeting JCAHO requirements for external databases by using PRO process. PMID- 8704662 TI - Effects of clinical supervision on nurse-patient cooperation quality: a controlled study in dementia care. AB - An intervention consisting of individualized planned care for patients and regular clinical systematic supervision for nurses was carried out on a ward for dementia care (the experimental ward, EW). On a similar control ward (CW), no changes were made. Observations of nurse-patient cooperation during morning care were collected at both wards, before and during the intervention. The observations (N = 107) were analyzed blindly and sorted into already-developed categories. The distribution of the cooperation-style categories was then analyzed statistically. The patients who survived throughout the study period (N = 18) were also compared over time. Nurse-patient cooperation at the EW improved significantly during the intervention period, whereas, at the CW, it deteriorated (p = .02). Status of surviving patients followed the same pattern (p = .001). The findings indicate that such interventions can improve the quality of nurse patient cooperation in dementia care. PMID- 8704663 TI - Comparison of two teaching strategies: adherence to a home monitoring program. AB - Two strategies for teaching lung and heart-lung transplant subjects to use an electronic spirometer/diary instrument at home to input and transmit daily respiratory, vital sign, and symptom measurements were compared. The first strategy integrated teaching into subjects' postoperative clinic visits, whereas the second strategy involved referral to a Patient Learning Center (PLC) for instruction. Adherence was increased by 30% when the teaching was transferred from the clinic setting to the PLC. These findings suggest that the PLC is an effective resource for preparing patients to manage their care after discharge from a hospital or clinic. PMID- 8704664 TI - Distribution of cues across assessed levels of infant pain. AB - A convenience sample of 24 more experienced, 20 less experienced, and 20 novice nurses assessed videotaped infants in varying degrees of pain (none, mild, moderate, and severe), as determined by an expert panel. Participants identified all information they used in making an assessment (All cues) and that subset they deemed most important (Key cues). Data were analyzed using a two-level analysis of variance (level of assessed pain and pediatric nursing experience). Forty-five of the 62 cues mentioned by participants did not differ across levels of pain. Eleven All cues and 11 Key cues differed between infants in pain and not in pain, suggesting that these cues may be potentially useful as predictors of the presence/absence of pain. Three All cues differed both between pain and no pain as well as between levels of pain, which suggests that participants' awareness of subtle leveling might be used in the discrimination of a wider range of pain levels. Differences in cue utilization among the more experienced, less experienced, and novice participants are discussed. PMID- 8704665 TI - After hospitalization: home health care for elderly persons. AB - The immediate post-hospitalization outcomes for elderly individuals who received home health services after discharge were compared with those who did not receive home health services. The needs of individuals with cognitive impairment and the "old-old" were major foci of interest. Assessment instruments included the Mini Mental State Exam, the Delirium Symptom Interview, and the Everyday Problem Solving Test. Data on the use of health services after hospital discharge were collected. Results showed that patients age 80 and older, who received skilled nursing services, were less likely to report health problems or complications than those who had not received skilled nursing services. Because the old-old elderly are more likely to have problems during the immediate post hospitalization period, this group of patients should be targeted for needs assessment and discharge planning. PMID- 8704666 TI - Participation of nurses in decision making for seriously ill adults. AB - The purpose of this study is to describe the involvement of nurses in the decision-making process of seriously ill hospitalized adults. Nurses (696) completed interviews with 1,427 patients. Patient, surrogate, and physician interviews were also completed. Patients and surrogates perceive the nurse as more influential in decision making than does the nurse or physician. Many nurses reported having no (31%) or little (36%) knowledge of their patients' preferences, and 53% of the nurses did not advocate for their patients' preferences. Only 50% of the nurses reported educating their patients about the treatment plan chosen or discussing treatment options with their patients, and few (17%) discuss prognosis. This study indicates nurses are not actively involved in the decision-making process of their patients, especially older or more experienced nurses and those working in intensive care units. PMID- 8704667 TI - Narratives by district nurses about elder abuse within families. AB - Twenty-one district nurses (DNs) narrated 44 cases of elder abuse within families. A phenomenological-hermeneutical analysis revealed that the experiences were complex and often included families providing care for an elderly person. The abuse seemed to be related to the inability of one party to meet the care demands required by the elderly, by him- or herself, or by the situation. It also seemed to be related to an inhibiting dependency between the parties, a negative execution of power over the weaker party, and a history of violence. When the experiences were considered within the perspective of Logstrup's ethics, it appeared that the DNs tried to remain neutral by not consciously reflecting on the ethical demands in the abuse situations. The findings suggest that nurses need support to enable them to decide about provision of care at home and to judge "the core" in abuse situations. They might gain this support through reflection on a meta-level, taking various perspectives into account, and thus making it possible to achieve new dimensions for decision making. PMID- 8704668 TI - Subjective appraisal of infant crying. AB - How families appraise difficult situations contributes to later adaptive functioning. We have observed in both research and practice that when appraising their infants' crying, mothers often compared their own infants' crying to actual or supposed much worse infants. They typically appraised their infants to be crying less than average infants. This phenomenon, defined in the social psychology literature, is called downward comparison. Secondary analysis of an established data set was used to assess the extent of downward comparison and the identification of excessive crying among a sample of 193 primiparous mothers. Mothers appraised their own infants' crying to be less than that of the expected average infant. Mothers who identified their infants as excessive criers made less robust downward comparisons than did mothers who did not identify their infants as excessive criers. Nursing interventions that focus on supporting mothers' positive appraisal and promoting knowledge of infant behaviors, particularly infant crying, are proposed. PMID- 8704669 TI - The effects of intravenous lidocaine in the prevention of postoperative sore throat. PMID- 8704670 TI - Hereditary angioedema: a potential clinical emergency. PMID- 8704671 TI - Anesthetic drug interactions. PMID- 8704672 TI - Physiological and practical considerations of fluid management. PMID- 8704673 TI - Ions and IVs: are we still using the Henry Ford model? AB - Intravenous fluid therapy matched to fluid losses provides the patient with appropriate and cost-effective care. While the physiognomies of many of our patients are robust enough to handle nearly any type of intravenous perioperative replacement, we shall all see patients with varying fluid and electrolyte disturbances. Using the tables and formulae in this article will assist the anesthesia provider in tailoring fluids to both tonicity and electrolyte needs. PMID- 8704674 TI - Fluid management for major surgical cases in the operating room. PMID- 8704675 TI - Fluid resuscitation. PMID- 8704676 TI - XXV Jubilee congress of the Polish Society of Phtysiopneumonology. Zakopane, November 5-8, 1995. Abstracts. PMID- 8704677 TI - Autotransplants for neuroblastoma. PMID- 8704678 TI - Recent results using CAMPATH-1 antibodies to control GVHD and graft rejection. AB - Recent data from the CAMPATH users group are reported. Different protocols have been tested using the CD52 antibodies Campath-1M and Campath-1G for prevention of GVHD and graft rejection in allogeneic transplants from both sibling and volunteer unrelated donors. Leukaemia relapse remains a significant problem for patients with CML, but in other diseases the recent results using T cell depletion appear to be as good as, or better than, published data with conventional GVHD prophylaxis. In addition, the morbidity and mortality associated with chronic GVHD are substantially reduced. Future collaborative studies to consolidate these findings include a randomised trial of the humanised antibody Campath-1H organised under the auspices of the EBMT. There are also plans to carry out experimental studies using Campath-1 antibodies to deplete T cells from peripheral blood stem cell harvests. PMID- 8704679 TI - Calcineurin protein phosphatase activity in peripheral blood lymphocytes. AB - The protein phosphatase activity of peripheral blood T lymphocytes (PBLs) was examined to quantify the contribution of calcineurin and other members of the family of serine/threonine protein phosphatases. Using selective phosphatase inhibitors, the fractional phosphatase activities of calcineurin, protein phosphatases 1 (PP1), 2A (PP2A), and 2C (PP2C) were determined. Okadaic acid was used to inhibit the activity of both PP1 and PP2A while cyclosporin A/cyclophilin or trifluoperazine were used as a specific inhibitors of the calmodulin-dependent phosphatase calcineurin. Using a [32P]labeled 19-residue phosphopeptide substrate, RII peptide, it was found that PP1 and PP2A comprise the majority of the total phosphatase activity in PBLs with okadaic acid inhibiting 80% of the phosphatase activity. The remaining 20% of the phosphatase activity can be attributed primarily to calcineurin since it was Ca2+ dependent, sensitive to inhibition by the calmodulin antagonist trifluoperazine, and inhibited by the complex of cyclosporin A (CsA) and cyclophilin. These results indicate that PBL extracts contain little PP2C activity. In addition, PBLs treated with CsA had measurably lower calcineurin activity in cell lysates. The measurement of calcineurin activity may provide a useful means of assessing the extent of immunosuppression during drug therapy. PMID- 8704680 TI - Phenotypic and functional characterization of peripheral blood and bone marrow natural killer cells prior to autologous transplantation. AB - NK cells can exert potent anti-leukemia activity after either autologous or allogeneic BMT. However, in autologous blood or marrow transplant patients, NK cell number and/or function could be reduced, and also may vary according to the sampling site. In order to evaluate the hypothesis that blood or marrow grafts from autologous transplant patients exhibit impaired NK cell activity that could contribute to disease recurrence, we evaluated the immunologic characteristics of NK cells in the bone marrow (BM) and peripheral blood (PB) from 27 patients undergoing autologous BMT, and also from 20 normal donors. We measured baseline and interleukin-2 (IL-2)-activated NK cell cytotoxicity, as well as expression of IL-2 receptors (IL-2R) (alpha-chain (p55) and beta-chain (p75)), and adhesion molecules. The cytotoxic activity of PB NK cells was significantly lower in autologous transplant patients than in normal donors (P < 0.0005) and this difference was not mitigated following IL-2 activation. In contrast, BM from autologous patients showed normal NK cell cytotoxicity, but contained higher numbers of NK cells (P < 0.025), with more intense CD56 expression (P < 0.05). Expression of p75 was lower on BM than on PB NK cells in both patients and normal donors. In addition, induction of p55 by IL-2 was abrogated in autologous PB NK cells. Therefore, depending on the site of harvest and the nature of donor cells (pre-BMT vs normal), our results show significant differences in NK cell number, function, and IL-2 receptor expression. This may affect relapse rates following autologous transplants performed with either PB or BM grafts. PMID- 8704681 TI - Representation of Hispanics in the National Marrow Donor Program. AB - Since less than one-third of patients in need of a BMT find related donors, most patients will rely on registries of volunteer donors. For patients from minority ethnic groups the chances of finding matched unrelated donors are lower, in part due to the smaller representation of minorities in the registries. Our purpose was to determine the representation of Hispanics in the National Marrow Donor Program (NMDP), the largest registry of volunteer marrow donors in the United States. We analyzed a database provided by the NMDP that contained information on minorities. The number of Hispanic volunteer donors has increased 110-fold in the last 6 years. The proportion of Hispanics in the registry has also increased from 1.1% to 6%. Nevertheless, the proportion of Hispanic patients that received unrelated marrow transplants facilitated by the NMDP has increased only from 2.8% to 3.9% since 1989. Only 19.7% of the formal searches initiated by Hispanic patients resulted in transplants compared to the 30.4% observed in the Caucasian population. Despite increments in the number and proportion of Hispanic volunteer donors, the proportion of Hispanics that receive BMT from unrelated donors remains low. We conclude that, in addition to increased recruitment efforts, other strategies will be necessary in order to find enough marrow donors to meet the needs of the Hispanic population. PMID- 8704682 TI - Toxicity of busulfan and cyclophosphamide (BU/CY2) in patients with hematologic malignancies. AB - Between July 1991 and January 1994, 52 patients with hematologic malignancies underwent BMT using BU/CY2 as conditioning regimen. Median patient age was 38 years. Eleven patients underwent autologous BMT, 22 HLA-identical allogeneic BMT, and 19 patients underwent a MUD or an allogeneic mismatched BMT. GVHD prophylaxis was with cyclosporine/methylprednisone in 26 patients; T cell depletion was used in 15 patients. VOD was observed in 7.5% of patients, IP in 12%, seizures in 4%. The overall incidence of grade II-IV acute GVHD was 35%. Delayed platelet engraftment was observed in seven of 11 patients who underwent autologous BMT. Graft failure was seen in seven of 19 (37%) patients who underwent MUD or allogeneic mismatched BMT. Six of the seven patients received T cell depletion as GVHD prophylaxis. BU/CY2 transplantation from an unrelated or family-mismatched donor with T cell depletion is associated with a high incidence of graft failure. PMID- 8704683 TI - High-dose carmustine, etoposide and melphalan ('BEM') with autologous stem cell transplantation: a dose-toxicity study. AB - We have investigated the toxicity of dose-escalation of BCNU, etoposide and melphalan ('BEM') chemotherapy with autologous stem cell transplantation in patients with haematological malignancies. Seventy-two patients with haematological malignancies were treated with BCNU (600 mg/m2, 450 mg/m2 or 300 mg/m2), etoposide 2 g/m2 and melphalan 140 mg/m2 followed by autologous bone marrow transplantation (ABMT), n = 51, or autologous peripheral blood progenitor cell transplantation (APBPCT), n = 21. Liver and pulmonary function was monitored pretransplant and at regular intervals post-transplant. Mucositis was graded daily during in-patient stay. There was a significantly higher incidence of symptomatic pulmonary toxicity in the patients who received BCNU at 600 mg/m2 than in the other two groups, and there was a significant increase in the incidence of asymptomatic decrease in carbon monoxide (KCO) in the patients who received BCNU 450 mg/m2. There was no significant difference between the three groups in the incidence and severity of mucositis or in the incidence of transiently abnormal liver function. We conclude that etoposide at 2 g/m2 can be used without unacceptable mucositis. BCNU at 600 mg/m2 is associated with an unacceptably high incidence of lung toxicity, but at 450 mg/m2 there is minimal symptomatic lung toxicity. PMID- 8704684 TI - Busulfan, cyclophosphamide and fractionated total body irradiation for allogeneic marrow transplantation in advanced acute and chronic myelogenous leukemia: phase I dose escalation of busulfan based on targeted plasma levels. AB - A previous phase I study determined that the maximum tolerated dose (MTD) of busulfan (BU) that could be given with a fixed dose of cyclophosphamide (CY) of 50 mg/kg and total body irradiation (TBI) dose of 12.0 Gy was 7 mg/kg. A phase II study was carried out in patients with advanced myeloid malignancies receiving allogeneic transplants without improvement in outcome as compared to historical controls. In that study, steady-state concentration (Css) of BU in 13 patients receiving a fixed dose of BU varied from 209 to 735 ng/ml. In an attempt to decrease the variability of the Css of BU, a study of targeting specific plasma concentrations was performed. In this study, BU dose was adjusted up or down based on first dose pharmacokinetics. The first dose level evaluated was 7.5 mg/kg with a target BU plasma level of 460 ng/ml. Six patients were entered at this level and the median BU plasma concentration achieved was 410 (range 390 533). One of six patients developed grade 3-4 regimen-related toxicities (RRT). Dose level II was a target of 559 ng/ml with a starting oral dose of BU of 9.6 mg/kg. Twelve patients were entered at this level and median plasma BU level was 548 (range 427-689). Three of 12 (25%) patients developed grade 3-4 RRTs and this was considered to be the MTD. The actuarial probability of grade II-IV acute GVHD was 0.70. Eight of 21 evaluable patients (38%) developed chronic GVHD. Of 18 patients who died, seven died of relapse at a median of 160 days (range 65-353) and 11 (61%) died of causes other than relapse at a median of 152 days (range 18 570). The actuarial probabilities of DFS, relapse and relapse-free mortality at 2 years in all patients were 0.13, 0.50 and 0.75, respectively. This study showed that targeted BU plasma levels within 10% of target can reliably be achieved with a bias of -2.07% and mean absolute error of 7.47%. Overall, targeting made a 31.8% to 100% in plasma BU Css as compared to expected BU Css based on first dose pharmacokinetics if targeting were not performed in this study. Thus targeting avoided much of the variability in BU concentrations seen in other studies. When compared with our previous phase II experience in same group of patients receiving same regimen, dose escalation of BU based on targeted plasma levels did not improve the outcome. PMID- 8704685 TI - Granulocyte recovery after sequential transfusion of mobilized blood stem cell concentrates in syngeneic recipients. AB - Three patients received sequential transfusions of G-CSF-mobilized peripheral blood stem cells from their identical twin in an attempt to abrogate neutropenia. Blood stem cells were harvested by leukapheresis in the healthy donor twins following mobilization with rhG-CSF at 5 micrograms/kg/day subcutaneously for at least 5 days. An average of 2.2 x 10(7) CFU-GM (range: 1.4-3.3) were collected and transfused without further manipulation. One patient, transfused with a CFU GM dose of 3 x 10(7) on day +6 after a syngeneic marrow transplant, experienced near-complete elimination of absolute neutropenia until spontaneous engraftment occurred on day +11. In the other two patients, we unexpectedly observed a transient granulopoietic inhibition, possibly related to the high T cell content of the blood stem cell transfusions. PMID- 8704686 TI - Close simulation of acute graft-versus-host disease by interleukin-2 administered after autologous bone marrow transplantation for hematologic malignancy. AB - The high relapse rate of hematologic malignancy treated with autologous bone marrow transplantation (ABMT) may reflect the absence of a graft-versus-leukemia (GVL) effect usually associated with graft-versus-host disease (GVHD). The purpose of this study was to determine whether administration of interleukin-2 (IL-2) early after ABMT might induce or exacerbate acute skin GVHD. Fourteen patients at high risk for post-transplant relapse, eight with NHL and six with AML > or = first relapse, were conditioned with chemotherapy and total body irradiation (13) or chemotherapy alone (1), and received purged (10) or unpurged (4) marrow. A median of 35 days (range 25-58) after ABMT, they received a 5-day induction course of Roche IL-2 (9 x 10(6) U/m2/day) followed by apheresis, reinfusion of LAK cells, and a 10-day maintenance course of IL-2 (0.9 x 10(6) U/m2/day), all by continuous i.v. infusion. Serial skin biopsies were obtained before and after IL-2 therapy and were read blindly. Patients were studied prospectively for the development of acute cutaneous GVHD as reflected by rash ( > or = 25% body surface area), skin biopsy ( > or = grade II histologic changes) and T cell infiltration as assessed by staining of the biopsy with antibodies UCHL-1 and TIA-1. No patient had a rash before IL-2 therapy, but 12 of 14 (85%) developed a rash during the IL-2 induction course. Before IL-2 therapy, biopsies from three of 10 patients (30%) revealed histologic GVHD; after induction IL-2, biopsies from 11 of 14 patients (79%) revealed grade II acute GVHD. Biopsies from all patients with histologic GVHD after IL-2 therapy contained TIA-1 positive T cells. HLA-DR was negative in the keratinocytes of these paraffin-embedded sections. One patient died early of sepsis, one patient required and responded to topical corticosteroids and 12 had spontaneous resolution of the rash. Six patients relapsed at 3-13 months, while seven remain in complete remission 32+ to 41+ months after ABMT. The results demonstrate that IL-2 therapy after ABMT can induce effects which histologically and clinically mimic cutaneous acute GVHD in most patients. Prospective, randomized trials of IL-2 vs observation after transplantation of autologous marrow or stem cells for high-risk NHL and AML have been initiated which may allow us to determine whether this phenomenon is associated with a clinical GVL effect as reflected by a decreased relapse rate. PMID- 8704687 TI - Allogeneic bone marrow transplantation in first remission for children with very high-risk acute lymphoblastic leukemia: a retrospective case-control study in the Nordic countries. Nordic Society for Pediatric Hematology and Oncology (NOPHO). AB - Among children with high-risk (HR) ALL there are subgroups with very-high-risk (VHR) features and poor prognosis despite developments in conventional chemotherapy for childhood ALL. We evaluated the outcome of VHR-ALL in children receiving allogeneic BMT (allo-BMT) in first remission (1CR) in a retrospective case-control study. In the population-based ALL material of the five Nordic countries, 22 children with VHR-ALL have undergone allo-BMT in 1CR between 1981 1991. We compared the outcome in these 22 children with 44 closely matched control patients who received conventional chemotherapy on HR-ALL protocols, as well as with a group of 405 children representing the remaining HR-ALL patients in the Nordic ALL database. The disease-free survival at 10 years was 73% in children receiving allo-BMT in 1CR, 50% in the matched controls (P = 0.02), and 59% in the remaining HR-ALL patients. The good prognosis of the allo-BMT group was due to a low relapse rate of 9%, as opposed to 41% in the group of matched controls. The superiority of allo-BMT as therapy in 1CR was mainly apparent in those with a very high WBC of > or = 100 x 10(9)/I at diagnosis; in the allo-BMT group 9/10 survived, as opposed to 8/20 of the matched controls (P = 0.03). We conclude that allo-BMT in 1CR should be seriously considered for children with a matched sibling donor and a VHR-ALL with WBC of > or = 100 and other established VHR criteria. PMID- 8704688 TI - Bone marrow transplantation for Philadelphia chromosome-positive acute lymphoblastic leukemia. AB - Between 1986 and 1995, 19 patients with Philadelphia chromosome-positive (Ph + ) acute lymphoblastic leukemia underwent 20 autologous (n = 9) or allogeneic (n = 11) blood or marrow transplant procedures in first (n = 12) or second (n = 3) remission, or in relapse (n = 5). Four patients died due to transplant-related causes, 11 relapsed at 3-39 months, one survives with disease which did not remit after transplant, and three are alive in continuous remission at 1, 26 and 65 months. Two of the relapsing patients are alive; one autografted patient after an allograft in second remission and one allografted patient after a donor leukocyte infusion. The projected overall survival is 37.5% at 3 years and 12.5% at 5 years. The 3-year probabilities of relapse and disease-free survival for autografted patients are 65.9% and 25.6% respectively, and for allografted patients, 63.4% and 21.8% respectively. The stage of the disease at the time of transplant or the type of transplant did not affect the outcome significantly, and late relapses beyond 3 years were seen after allogeneic as well as autologous transplantation. In our experience, the outcome of patients with Ph + acute lymphoblastic leukemia continues to be poor despite high-dose therapy due to high relapse rates, and the development of additional measures to enhance the antileukemic efficacy of bone marrow transplantation is necessary. PMID- 8704689 TI - Clinical and hematologic response of chronic lymphocytic and prolymphocytic leukemia persisting after allogeneic bone marrow transplantation with the onset of acute graft-versus-host disease: possible role of graft-versus-leukemia. AB - One patient with refractory B cell chronic lymphocytic leukemia (CLL) and another with refractory B cell prolymphocytic leukemia (PLL) underwent bone marrow transplantation (BMT) from HLA-identical siblings. Circulating malignant cells persisted at high levels in the patient with PLL and there was clinical evidence of disease progression soon after transplant in the patient with CLL. Starting 4 5 weeks post-BMT, cyclosporine was tapered rapidly to stimulate immunologic graft versus-leukemia (GVL) reactions. There was a fall in the number of malignant cells and reversal of organomegaly with the onset of acute graft-versus-host disease (GVHD). Both patients received conventional doses of corticosteroids for GVHD which also may have contributed to disease response to some extent. Total clearance of the leukemic cells from the peripheral blood was seen in both patients, and clearance of the marrow was seen in the patient with CLL. However, both patients died of complications of severe GVHD. We conclude that GVHD may be associated with a GVL effect after allogeneic BMT for refractory chronic B cell lymphoproliferative diseases. Whether GVL reaction occur in the absence of clinically obvious GVHD after allogeneic BMT for CLL remains to be seen. PMID- 8704690 TI - Immunomagnetic CD4+ and CD8+ cell depletion for patients at high risk for severe acute GVHD. AB - Acute GVHD remains a major problem in allogeneic BMT, in particular when donors other than HLA-identical siblings are used. To determine the efficacy of an immunomagnetic method for depletion of CD4+ and CD8+ lymphocytes from the marrow graft, a series of 15 patients was studied. Thirteen patients had matched unrelated donors, and two patients had related donors. Cyclosporine was used as GVHD prophylaxis in combination with CD4+ and CD8+ depletion, which removed 94.1 +/- 3.2%, 97.0 +/- 5.1%, and 96.7 +/- 3.1% of CD3+, CD4+ and CD8+ cells, respectively. All patients engrafted promptly with AGC > 500/mm3 after a median of 16 days post-BMT. Acute GVHD grade II-IV developed in 0/2 related transplants and 4/13 MUD transplants; only one patient had grade III-IV acute GVHD. No late graft failure was observed. Three patients relapsed; two had advanced disease at the time of BMT. Seven patients are alive and in CCR after a median of 497 days; actuarial survival is 39% at 24 months. The fever syndrome observed with selective CD8+ cell depletion was not seen with the combined CD4+ and CD8+ cell depletion. Immunomagnetic CD4+ and CD8+ cell depletion of marrow grafts, in combination with in vivo cyclosporine, is a simple, reproducible and effective method to decrease the incidence and severity of acute GVHD in patients at high risk for this complication after allogeneic BMT. PMID- 8704691 TI - High lactate dehydrogenase level is associated with an adverse outlook in autografting for Hodgkin's disease. AB - Forty-two patients with relapsed or refractory Hodgkin's disease (HD) were treated with high-dose chemotherapy (BEAM regimen) followed by autologous bone marrow and/or peripheral blood progenitor cell (PBPC) rescue. There was one procedure-related death and the overall response rate at 6 months was 88% (95% confidence interval 78-98%). The 2 year overall and event-free survival was 81% (95% confidence interval 65-96%) and 74% (95% confidence interval 58-89%) respectively. Median follow-up was 33 months. The use of PBPC instead of marrow resulted in a significant shortening of the time to engraftment (P < 0.01). Multivariate analysis identified the pre-transplant LDH level as a highly significant factor in predicting overall survival (P = 0.007). The BEAM regimen is an effective conditioning schedule that is well tolerated but patients with a raised LDH at the time of transplant remain at high risk of early relapse and death due to disease. PMID- 8704692 TI - High-dose chemotherapy with autologous marrow rescue for malignant brain tumors: analysis of the impact of prior chemotherapy and cranio-spinal irradiation on hematopoietic recovery. AB - The relative impact of age, sex, nucleated cell dose, prior chemotherapy, prior cranio-spinal irradiation (CSI) and bone marrow harvest (BMH) site on hematological recovery after ABMT were analyzed in a multivariate model. The study population comprised 100 patients with a median age of 9 years who underwent ABMT for malignant brain tumors. Two engraftment parameters were evaluated: number of days post ABMT before (1) an absolute neutrophil count (ANC) > or = 0.5 x 10(9)/l and (2) a platelet count > or = 50 x 10(9)/I were achieved for the third consecutive day without transfusions. Increasing cell dose correlated significantly with a more prompt recovery of platelet counts and ANC. Previous chemotherapy significantly delayed both neutrophil and platelet engraftment. The group of patients who also received CSI had a very delayed platelet recovery with a median time to engraftment of 72 days. Neutrophil engraftment was also significantly delayed and occurred at a median of 23 days. This effect of CSI was independent of cell dose or prior chemotherapy. In 20 of these patients, marrow was harvested at least partially from the posterior iliac crests, which might have received significant doses of irradiation. We conclude that engraftment is significantly faster if bone marrow is harvested prior to any chemotherapy administration, and that patients who receive prior CSI may have significant engraftment delay, particularly of the platelet lineage. In this latter group of patients, marrow should not be harvested from the posterior iliac crests. Strategies that might enhance both neutrophil and platelet count recovery should be considered in patients with irradiation damage to a substantial proportion of the total hematopoietic tissue. PMID- 8704693 TI - Bone density loss during treatment of chronic GVHD. AB - Nine adult patients 31-47 (median 39) years of age treated with prednisone and cyclosporin A (CsA) for chronic graft-versus-host disease (GVHD) were evaluated for biochemical factors associated with skeletal turnover at initiation of immunosuppressive therapy (3 months after marrow transplant) and 9 months later (follow-up). Absorptiometry studies of the wrist and lumbar spine were also performed. Serum levels of 1,25-dihydroxycholecalciferol (1,25(OH)2D) were decreased at enrollment, particularly in the six males. Values for all nine patients remained low at follow-up. Levels of serum 25-hydroxycholecalciferol (25(OH)D), parathyroid hormone, and ionized calcium were normal at enrollment and follow-up. Mean urine hydroxyproline and calcium levels were elevated at enrollment, suggesting increased bone resorption; the mean values decreased to the high normal range at follow-up. Urine magnesium excretion was elevated in eight of nine patients at baseline and remained elevated at follow-up in three of eight evaluable patients. Single and dual photon absorptiometry of the wrist and spine, respectively, and dual energy X-ray absorptiometry of the spine, were utilized to evaluate bone mineral density over time. The precision of these tests was, respectively, +/- 3.5%, +/- 3.1% and +/- 1.0%. Results showed a significant ( > 2.5 times the precision) decrease over 9 months in bone mineral density in three of five evaluable males and all three females. The findings indicate increased collagen and bone turnover, increased urinary magnesium and calcium excretion and a significant risk of osteoporosis in patients receiving treatment for chronic GVHD. Preventive measures, including gonadal hormone replacement in females, should be initiated early after transplantation. Further studies are needed to identify patients at highest risk of bone loss and to monitor the effects of preventive therapy. PMID- 8704694 TI - Schistosomiasis as a predisposing factor to veno-occlusive disease of the liver following allogeneic bone marrow transplantation. AB - Among 89 allogeneic bone marrow transplant recipients, veno-occlusive disease of the liver (VOD) was diagnosed in 10 patients (11.2%). All cases (n = 5) with schistosomal hepatic periportal fibrosis detected by pretransplant ultrasonography, developed severe fatal VOD in spite of normal initial liver functions and absence of portal hypertension. The incidence of VOD among patients without previous schistosomal contact was 5.95% (5/84). The relative risk to develop VOD was calculated to be 16.8-fold higher in patients with previous schistosomiasis. Schistosomal hepatic periportal fibrosis may thus be added to the known risk factors predisposing to the development of VOD in allogeneic transplant recipients. PMID- 8704695 TI - Neurologic complications following bone marrow transplantation for sickle cell disease. AB - A boy with sickle cell anemia underwent bone marrow transplantation (BMT). He was normal on neurological examination, but had radiologic evidence of an old left frontal lobe infarct, multiple cerebral vascular stenoses and moyamoya collaterals. After BMT he developed seizures with extension of the infarct and subarachnoid hemorrhage. One year later angiography revealed worsening stenosis of the M1 segments of both middle cerebral arteries. At that time an increase in von Willebrand's factor with decreased large molecular weight multimers (LvWF) was observed. We speculate that LvWF dependent, shear-induced platelet aggregation, together with endothelial damage may have contributed to the development of neurologic complications in this patient. PMID- 8704696 TI - Mucormycosis after bone marrow transplantation: report of four cases in thalassemia and review of the literature. AB - We report four cases of mucormycosis that occurred among 711 patients who underwent BMT for thalassemia, and review 18 additional cases among BMT recipients that were reported in the English-language literature. All these patients were polytransfused and were in advanced phase of disease with severe acquired hemochromatosis. The sites of infection were sinonasal, rhinocerebral pulmonary, pulmonary and pulmonary-central nervous system. Mucormycosis was the primary cause of death in three of four patients. Two infections were detected within the first 100 days after BMT. Only one of the four patients had partial resolution of sinonasal mucormycosis following aggressive antifungal therapy combined with hyperbaric oxygen treatment. PMID- 8704697 TI - Prolonged deficiency of protein C following hematopoietic stem cell transplantation. AB - Deficiencies in circulating anticoagulant protein C (PC) occur in patients undergoing hematopoietic stem cell transplantation. These deficiencies may predispose to thrombotic and other complications, and may contribute to the morbidity of transplantation. In most patients, PC reaches a nadir 14 days after the preparative regimen and then begins to increase toward normal. However, low PC has been seen months after transplantation. Neither the frequency of, nor risk factors for, this prolonged deficiency are known. We examined 71 adults undergoing stem cell transplantation and found low PC antigen and activity in 21% and 20% of patients, respectively. Low PC at day 100 correlated strongly with low PC pre-chemotherapy (PC antigen, r = 0.69, P < 0.001; PC activity, r = 0.59, P < 0.001). The incidence of deficiency of PC at day 100 was lower in patients undergoing peripheral stem cell transplantation compared with patients undergoing autologous BMT (12.5% vs 35%; P = 0.05), although several significant confounding variables exist. We conclude that deficiencies in protein C persist at least 100 days after stem cell transplantation in nearly one quarter of patients undergoing this procedure. Therefore, patients undergoing stem cell transplantation may be at prolonged risk of thrombotic and other complications. Further studies to determine the risk of prolonged deficiency based on stem cell source need to be performed. PMID- 8704698 TI - Quantitative determination of immunological components of salivary gland secretion in transplant recipients. AB - Cancer therapies often induce oral complications. We studied longitudinally the major salivary immunoglobulin content in 42 transplant recipients using an enzyme linked immunosorbent assay. A dramatic decrease in both sIgA and IgG is noticed post-transplant compared to pretransplant values. Only a small recovery of these immunoglobulin concentrations is seen after approximately 4 months. The reduction of sIgA and IgG in saliva could contribute to the frequent oral complications seen in these patients after transplantation and increase the risk of developing dental caries. PMID- 8704700 TI - G-CSF in an infant donor: a method of reducing harvest volume in bone marrow transplantation. AB - We report the case of a 4-year-old female with high-risk ALL in first CR who received a BMT from an 11-month-old matched sibling treated with G-CSF in order to obtain an adequate number of mononuclear cells in a limited volume of bone marrow. The absence of toxicity, efficacy of the procedure and quality of the post-transplant clinical outcome suggest such treatments are feasible and useful to overcome problems caused by donor age and/or body weight. In view of this experience we demonstrate how such an approach leads to a notable reduction in risks and in bone marrow donation costs. PMID- 8704699 TI - Long-term cryopreservation of bone marrow for autologous transplantation. AB - Little is known about the effect of long-term cryopreservation on the viability of hematopoietic stem cells (HSC) or on the success of autologous bone marrow transplantation. Although progenitor cell assays such as culture of CFU-GM after thawing can be predictive of engraftment, the most rigorous assay for the cryosurvival of HSC is engraftment after reinfusion of stem cells. We retrospectively evaluated the engraftment data for 36 patients with hematologic malignancies or solid tumors treated at the Fred Hutchinson Cancer Research Center between 1981 and 1993 who received bone marrows stored for 2 years or more. The median duration of cryopreservation for this study group was 2.7 years (range 2.0-7.8). Ninety-seven percent of patients in the study group achieved a granulocyte count of > or = 0.5 x 1.0(9)/1 at a median of 19 days (range 10-115) vs 86% of control group (selected by diagnosis and date of storage) at a median of 20 days (P = 0.14). Seventy percent of patients in the study group achieved a platelet count > or = 20 x 10(9)/1 at a median of 27 days (range 9-69) vs 74% of control group at a median of 23 days (P = 0.47). Also, samples of 28 marrows cryopreserved for a median of 4.4 years (range 2.0-7.8) were cultured to determine if a loss of hematopoietic progenitors relative to duration of storage could be detected. The storage length was not predictive for the quantity of colonies formed (P = 0.57 for BFU-E-derived colonies; P = 0.65 for CFU-GM-derived colonies). We found no consistent detrimental effect of long-term cryopreservation on the success rate of autologous bone marrow transplantation. This report confirms previous reports that marrow cells cryopreserved for several years are capable of engrafting. Therefore, bone marrow cells may be stored at an early appropriate time before the side-effects of multiple cycles of chemotherapy and radiotherapy on hematopoietic tissues are incurred. PMID- 8704701 TI - Acute iritis induced by granulocyte colony-stimulating factor used for mobilization in a volunteer unrelated peripheral blood progenitor cell donor. AB - We describe a volunteer unrelated peripheral blood progenitor cell donor with previously diagnosed dermatitis herpetiformis in whom the administration of G-CSF for the mobilization of precursor cells induced acute iritis. G-CSF has been administered to healthy people with minimal side-effects but when used in patients with autoimmune disorders worsening of symptoms or new manifestations may be a potential concern. PMID- 8704702 TI - Hematopoietic clone with karyotypic abnormalities of host origin after bone marrow transplantation: two case reports. AB - A patient with non-Hodgkin's lymphoma in remission developed a myelodysplastic syndrome (MDS) 12 years after ABMT. This patient had undergone bone marrow harvesting prior to any chemoradiotherapy. He had received the autograft following conditioning with high-dose CY and TBI. Chromosomal analysis of BM cells revealed complicated abnormalities. Similar karyotypic abnormalities in host-derived BM cells were found in another patient with AML who had received allogeneic BM following conditioning with CY plus TBI 15 months previously. These findings suggest that MDS or clonal karyotypic abnormalities following ABMT may derive from endogenous hematopoietic stem cells that survive the BMT preparative regimen. PMID- 8704703 TI - Donor leukocyte infusions for treatment of relapsed acute myeloid leukemia after allogeneic bone marrow transplantation. AB - A 24-year-old man with acute myelomonocytic leukemia (AML-M4) who relapsed 6 months after an allogeneic BMT was treated with chemotherapy followed by donor leukocyte infusions (4.19 x 10(8) mononuclear cells/kg). The patient developed grade II acute GVHD that responded to therapy with CsA and prednisone. Chimerism was assessed by PCR amplification of the MCT 118 hypervariable region. Fourteen months after donor leukocyte infusions the patient remains in complete remission, without any morphologic and cytogenetic evidence of leukemia, and with a complete donor chimerism. This case shows that donor leukocyte infusions are an effective therapy for some acute myeloid leukemia patients who relapse after allogeneic BMT. PMID- 8704704 TI - Treatment of veno-occlusive disease of the liver with bolus tissue plasminogen activator and continuous infusion antithrombin III concentrate. AB - Veno-occlusive disease (VOD) of the liver is a common complication of BMT and is accompanied by reduced levels of natural anticoagulants and by multi-organ dysfunction. We describe two cases of clinical VOD developing after autologous BMT and accompanied by ultrasonographic features of reversed portal venous flow. In both cases the patients had decreased levels of antithrombin (AT). Once the diagnosis of VOD was made, these patients were treated with tissue plasminogen activator (tPA) and continuous infusion AT. Each patient had radiographic and clinical resolution of VOD with the therapy. This novel treatment appears to have reversed the course of VOD without the increased risk of bleeding seen in the use of heparin therapy. PMID- 8704705 TI - Graft-versus-host reaction spares normal stem cells in chronic myelogenous leukemia. AB - Chronic myelogenous leukemia is a clonal proliferative disorder of pluripotent hematopoietic stem cells. Cure may be achieved by myeloablative conditioning treatment and marrow transplantation. In addition, allogeneic marrow can exert a graft-versus-leukemia effect. The graft-versus-leukemia effect may be directed against leukemia-specific antigens or against antigens on all hematopoietic cells, or it can be part of a graft-versus-host reaction. We report an informative post-transplant course of a patient with yet another leukemia specific effect. This patient was transplanted with marrow from his HLA-identical sister in an advanced phase of CML and developed acute and chronic GVHD. After a severe pneumonia a high proportion of his metaphases in the bone marrow were male and Philadelphia chromosome negative. Later all metaphases were again female and leukemic cells could not be detected by reverse transcriptase polymerase chain reaction analysis (RT-PCR) for BCR/ABL. This course indicates that normal hematopoietic stem cells may survive intensive chemotherapy, bone marrow transplantation and GVHD. They may be recruited from a dormant state into proliferation during severe infections. In contrast, CML may be eliminated by the graft-versus-host reaction that recognizes recruited cells and spares dormant cells. PMID- 8704706 TI - Reconstitution with Philadelphia chromosome-negative recipient haematopoiesis early after allogeneic BMT for CML. AB - We report a 33-year-old man with Ph chromosome-positive CML who underwent an allogeneic BMT from an unrelated donor. DNA microsatellite studies showed complete donor chimaerism immediately after BMT followed by mixed chimaerism; by day + 45 haematopoiesis was exclusively of recipient origin. Throughout the first year post-transplant all marrow metaphases were Ph negative but with non-clonal rearrangements consistent with autologous recovery. Cytogenetic relapse of leukaemia was first detected 15 months post-transplant. This case is unusual in that non-malignant stem cells of recipient origin survived the transplant and reconstituted haematopoiesis very early after BMT. Later the leukaemic cells reasserted their 'proliferative' advantage. PMID- 8704707 TI - Autologous red cell recovery and relapse of multiple myeloma after bone marrow transplantation: is this a graft-versus-myeloma effect? AB - We report a patient who, following an allogeneic bone marrow transplant for multiple myeloma, recovered autologous erythropoiesis which was rapidly followed by relapse of her multiple myeloma. We postulate that the loss of the graft (as demonstrated by loss of donor erythropoiesis) and subsequent relapse of the multiple myeloma may be support for the existence of a graft-versus-myeloma effect. PMID- 8704708 TI - Successful paternity of twins following bone marrow transplantation with busulfan, melphalan and cyclophosphamide conditioning. AB - A 33-year-old man who had received previous chemotherapy with cytarabine, daunorubicin and mitoxantrone followed by an autologous marrow transplant after conditioning with busulfan, melphalan and cyclophosphamide, fathered sex mismatched fraternal twins approximately 6 years post-transplant. HLA and DNA analyses showed the probability of paternity to be in excess of 99% for each twin. To our knowledge this represents the first documented case of paternity following conditioning with this combination of marrow ablative agents and the first report of twin paternity following autologous marrow transplantation. PMID- 8704709 TI - Long-term remission obtained by intensive chemotherapy against a leukemic relapse and subsequent graft-versus-host disease following an allogeneic BMT in a patient with myelodysplastic syndrome. PMID- 8704710 TI - Grade IV graft-versus-host disease and a 1-year remission following donor leukocyte transfusion for treatment of relapsed acute lymphoblastic leukemia. PMID- 8704711 TI - Significance of CMV DNA PCR results in bone marrow transplant recipients. PMID- 8704712 TI - Successful pregnancy after allogeneic bone marrow transplantation following conditioning including a 10-Gy single exposure total body irradiation. PMID- 8704713 TI - Cytokine mobilized blood CD34-expressing cells are not cycling. PMID- 8704714 TI - The HelpAge International: global feminisation of care. PMID- 8704715 TI - Chronic renal failure in old age. PMID- 8704717 TI - Clinical criteria help diagnose brain disorder. PMID- 8704716 TI - Venous and arterial leg ulcers (continuing education credit). PMID- 8704718 TI - Nursing homes: lest we forget. PMID- 8704719 TI - A praiseworthy purpose in the UK. Interview by Pat Young. PMID- 8704720 TI - Treating osteoporosis. PMID- 8704721 TI - Self-starvation through the ages: reflections on the pre-history of anorexia nervosa. AB - Recent publications have indicated that voluntary self-starvation is not a recently developed syndrome and that it has been reported throughout history. These prior forms of inedia are summarized and related to their historical and cultural contexts. On the basis of these data, some hypotheses are proposed regarding social influences on the vulnerability to eating disorders. OBJECTIVE: To document and describe forms of eating disorders occurring prior to the formal medical description of anorexia nervosa in the late 19th century. METHOD: Review of historical references to self-starvation, of recent publications on the history of eating disorders, and of articles describing cases of eating disorders occurring in the past. RESULTS: Forms of eating disorders have existed since ancient times varying in frequency, manifestations, and possible motivation. DISCUSSION: Certain sociocultural factors appear to foster or inhibit the frequency and type of eating disorders. PMID- 8704723 TI - Issues and concerns associated with different risk models for eating disorders. AB - OBJECTIVE: The present paper examines issues and concerns associated with different risk models in identifying individuals who may be vulnerable for eating disorders. METHOD: Studies were located by computerized search and the authors' knowledge of the literature. For the purposes of this paper, different risk models are grouped according to three types of sample selection criteria: exposure to environmental pressures toward thinness, parental psychopathology, and intraindividual characteristics. Intraindividual characteristics were subdivided into biological and behavior vulnerability markers, and within the behavior risk approach, symptom and nonsymptom risk models were further identified. RESULTS: Our literature review indicates that risk research on eating disorders is still in its formative years. More well-planned prospective risk studies are needed. CONCLUSIONS: Among these risk models, the nonsymptom risk approach, which defines risk on the basis of nonsymptom vulnerability markers, represents one of the more promising avenues for future risk research and deserves further exploration. PMID- 8704722 TI - Eating disturbances among American minority groups: a review. AB - OBJECTIVE: The purpose of this paper was to review the research literature related to eating behaviors and disturbances among American minority groups. METHOD: A computer-based literature search was conducted to locate articles pertaining to this topic. RESULTS: This review indicates that, compared to Caucasian females, eating disturbances are equally common among Hispanic females, more frequent among Native Americans, and less frequent among Black and Asian American females. Risk factors for eating disorders (EDs) are greater among minority females who are younger, heavier, better educated, and more identified with White, middle-class values. DISCUSSION: Further studies of EDs among American minority groups are needed, especially studies that are longitudinal and developmental in nature and that focus more specifically on the effects of racism in the development of EDs. PMID- 8704724 TI - Cross-cultural differences in maternal evaluation of children's body shapes. AB - OBJECTIVE: This study investigated cultural variation in mothers' attitudes to children's body shape. METHOD: One hundred thirty-one mothers from five cultural groups attending a pediatric clinic were approached, and data obtained from 114. Background information was obtained, including weight and height for themselves and their children. Mothers completed the Eating Attitudes Test (EAT-26) and rated drawings of children for attractiveness and health, using Likert scales, scored 1-7. RESULTS: Mothers from the different cultural groups had similar body mass index (BMI) and EAT scores, and their children had similar average body weight. However, UK mothers found slimmer girls attractive compared to mothers from South Asia, the Mediterranean or the Caribbean regions, and sub-Saharan Africa (p < .05). The differences occurred within the mid-range (median scores for all ethnic groups 4-5). South Asian mothers presented to the pediatric clinic with more worries about their children not gaining weight and growth (p < .01). DISCUSSION: These findings have implications for understanding cultural variation in the acquisition of attitudes to body shape, and these attitudes influence medical help seeking. PMID- 8704725 TI - Body shape ideals across gender, sexual orientation, socioeconomic status, race, and age in personal advertisements. AB - OBJECTIVE: To assess body shape ideals across gender, sexual orientation, race, socio-economic status, and age, METHOD: An analysis of personal advertisements was conducted across seven different publications which targeted the groups of interest. RESULTS: Women advertised body weight much less often than men, and lesbians reported body shape descriptors significantly less often than heterosexual women. Gay men and African-American men described their body shape significantly more often than did other groups. However, their reported body mass indices (BMI) were significantly different-African-American men reported a higher BMI, and gay men a lower BMI, than Euro-American heterosexual men. DISCUSSION: Race and sexual orientation may influence the importance of size of body shape ideals for men. For women, however, their advertised weights conformed to the thin ideal across all groups surveyed. Gender roles affecting body shape ideals and mate attraction are discussed. PMID- 8704726 TI - Racial differences in body type preferences of men for women. AB - OBJECTIVE: Research indicates that African-American women have a significantly higher prevalence of obesity, a markedly lower prevalence of eating disorders, and greater satisfaction with their bodies than Euro-American women. One potential contributing explanation for this difference may be differential body type preferences between the men in the two communities. METHOD: Sixty-three African-American and 116 Euro-American men were asked to rank, in order of attractiveness, a series of silhouettes of women of varying sizes. Additionally, they answered questions concerning their current relationships. RESULTS: Euro Americans chose significantly thinner figures, and reported wishing their girlfriends would lose weight significantly more often than African-Americans. DISCUSSION: These differential preferences may translate into greater pressure within the Euro-American community for women to be thin than in the African American community; however, a small effect size suggests that factors other than race contribute to men's body type preferences. PMID- 8704727 TI - Rapid responders to intensive treatment of bulimia nervosa. AB - OBJECTIVE: The purpose of the study was to examine patterns of response to treatment in bulimia nervosa and to consider the utility of differentiating rapid from slower responders. METHOD: Participants were 166 female patients with bulimia nervosa who received specialized day hospital treatment for their eating disorder and provided complete data on symptom frequencies over the course of treatment. Symptoms and psychological functioning were assessed at the beginning and end of treatment and 2-year follow-up information was available for a subsample of 57 patients. RESULTS: A large subgroup (41%) of patients responded rapidly to treatment (i.e., symptom frequencies of three or less during the first 4 weeks of treatment) and a smaller subgroup (31%) were considered slower responders (i.e., symptom frequencies of four or more over the first 4 weeks of treatment and three or less over the last 4 weeks of treatment). The remaining patients were either partial responders (18%) or nonresponders (10%). Rapid responders tended to be older, less symptomatic, and less preoccupied with binging before treatment (all ps < .05), but the differences were not strong. Rapid responders also had better symptom control at the end of treatment (p < .00001), were less likely to receive antidepressant/antibulimic medication during the program (p < .04), and were significantly less likely to relapse within 2 years of attending the program (p < .005). DISCUSSION: These findings demonstrate that a significant subgroup of severely ill bulimia nervosa patients had a rapid, strong, and enduring response to intensive treatment, but could not be well identified with the available measures before treatment. PMID- 8704728 TI - Is bulimia nervosa increasing in frequency? AB - OBJECTIVE: To assess whether or not claims of an increase in the incidence of bulimic disorders are empirically justified. METHOD: Review of the epidemiological data informative to gauge time trends: Studies based on hospital records or registers, community surveys of bulimic disorders, questionnaire-based investigations. RESULTS: Epidemiological studies conducted since 1980 do not show an upward trend in rates. Reports of increased rates among recent birth cohorts were unreliable and did not control for confounding comorbid disorders. Changes in diagnostic and referral practices are likely to account for higher numbers of patients seen in specialized treatment centers. DISCUSSION: Although based on a limited number of studies, the empirical evidence does not support secular changes in the incidence of bulimic disorders. In keeping with this conservative conclusion, it is noted that high rates of dieting and body dissatisfaction were already reported 30 years ago among adolescent girls. PMID- 8704729 TI - Development and preliminary validation of the cognitive behavioral dieting scale. AB - OBJECTIVE: The purpose of this research was to empirically develop the Cognitive Behavioral Dieting Scale (CBDS), a measure of current dieting. METHOD: The first study involved item generation and a procedure to boost internal consistency while reducing scale length. Study 2 involved a factor analysis and measures of scale reliability. The third study evaluated the ability of the CBDS to predict calorie intake and negative calorie balance from a 24-hr diet recall. Study 4 evaluated construct validity by comparing the CBDS to dietary restraint, body image, and health behavior self-efficacy. RESULTS: The CBDS is a 14-item scale which measures current dieting behavior and related thoughts within the past 2 weeks. Internal consistency was alpha = .95 and 2-day test-retest reliability was r = .95. This scale provides a method for operationalizing dieting, provides a construct that is different from restraint, and assess dieting behavior on a continuum. Additionally, this scale was able to predict calorie intake and negative calorie balance above and beyond the predictive ability of physical variables (i.e., body mass index BMI] and exercise calories). An additional study of construct validity showed the CBDS was related to poor body image esteem and dietary restraint, but minimally related to healthy eating self-efficacy. DISCUSSION: In conclusion, the CBDS shows promise as a valid and reliable measure of dieting behavior. This scale should have utility in future research on how current dieting relates to eating disorders, dietary restraint, and obesity. PMID- 8704730 TI - The reliability of bioelectrical impedance analysis for measuring changes in the body composition of patients with anorexia nervosa. AB - OBJECTIVE: To determine whether bioelectrical impedance analysis (BIA) is a valid measure of change in body fat in anorexia nervosa (AN) patients during refeeding, as compared to skinfold calipers (SF). METHODS: Prospective cohort study with measures of BIA and SF performed once a month for 3 months on patients meeting the DSM-III-R criteria for AN who received treatment from the Eating Disorders Clinic of a university teaching hospital as inpatients or outpatients. RESULTS: Twenty patients completed the study. Comparison of the two methods by the Pearson correlation coefficient showed a weak negative correlation of r = -.305. Analysis by a graphical method confirmed the poor agreement that exists between the two methods. DISCUSSION: The inability of BIA to detect changes in body composition due to altered hydration, and to accurately assess the distribution of water between intracellular and extracellular compartments, limits its clinical usefulness in AN. It appears that SF measurements are preferable to BIA as a measure of body fat change in patients with AN. PMID- 8704731 TI - Morphological evidence for a galanin-opiate interaction in the rat mediobasal hypothalamus. AB - It is well established that hypothalamic galanin- and beta-endorphin-containing circuits play important roles in the neuroendocrine regulation of pituitary hormone secretion and sexual behaviors, as well as in feeding. Recent experimental evidence suggests that an opiate-galanin interaction may be involved in these neuroendocrine responses. In particular, galanin and beta-endorphin have been shown to stimulate prolactin release from the pituitary, and concurrently, evoke feeding in the rat. The present study was designed to elucidate the morphological component underlying these responses in the hypothalamus. Sections of the mediobasal hypothalamus of colchicine-pretreated female rats were double immunostained for galanin and beta-endorphin. A dark blue nickel ammonium sulfate intensified diaminobenzidine reaction was used to visualize galanin profiles, while beta-endorphin neurons were labeled with a light brown diaminobenzidine reaction. Light microscopy revealed putative connections between galanin boutons and beta-endorphin cells. Electron microscopic examination showed that galanin boutons form axo-somatic and axo-dendritic synaptic connections with beta endorphin neurons. The vast majority (89.6%) of the beta-endorphin-immunoreactive neurons were found to be contacted by galanin-immunopositive fibers in the hypothalamus. To determine the origin of the galanin fibers innervating this region, the arcuate nuclei of additional rats were isolated unilaterally using a Halasz-knife. After a ten day survival period, immunostaining was carried out for galanin. The relative surface occupied by galanin immunoreactive profiles on the ipsi- and contralateral sides were compared using an image analyzer. This analysis revealed that deafferentation of the arcuate nucleus did not decrease the density of galanin immunoreactive profiles on the isolated side of the arcuate nucleus compared to the control side, thus, indicating that the galanin boutons contacting beta-endorphin cells are most probably of local origin. These studies support the proposal that galanin-evoked prolactin secretion and feeding behavior may, in part, be mediated by enhanced beta-endorphin release and raises the possibility that a hypothalamic galanin-beta-endorphin axis may operate in the control of other pituitary hormones. PMID- 8704732 TI - Effects of dehydroepiandrosterone (DHEA) on pituitary prolactin and arcuate nucleus neuron tyrosine hydroxylase mRNA levels in the rat. AB - It is well documented that dehydroepiandrosterone (DHEA), an adrenal androgen, is converted into potent androgens and/or estrogens in peripheral tissues. Since sex steroids are involved in the regulation of prolactin (PRL) secretion, we have studied the effect of DHEA administration on PRL mRNA levels in both adult male and female rats. Since tuberoinfundibular dopaminergic (TIDA) neurons are involved in the negative regulation of PRL, we have also evaluated the effects of DHEA on the genetic expression of tyrosine hydroxylase (TH), the limiting enzyme in catecholamine biosynthesis in TIDA neurons. Sham-operated and castrated animals of both sexes received during 2 days DHEA at the dose of 6 mg/kg/day, starting on the first day after castration. PRL and TH mRNA levels were measured by quantitative in situ hybridization. In the male rat, orchiectomy performed 3 days earlier did not modify PRL mRNA levels. DHEA administration increased the hybridization signal in both sham-operated and orchiectomized animals. In the female, ovariectomy decreased PRL mRNA levels and, as observed in the male, DHEA treatment induced an increase in the hybridization signal in both control and ovariectomized rats. In TIDA neurons, castration increased TH mRNA levels as evaluated by number of grains over labelled neurons and the number of TH-labelled cells per section in both male and female animals. In both sham-operated male rats and orchiectomized animals, DHEA decreased the hybridization signal. In the female, DHEA administration completely prevented the increase in TH mRNA levels due to ovariectomy. In sham-operated female rats, the treatment had no effect. These data clearly indicate that in both male and female rats DHEA exerts an estrogenic influence on both PRL and TH gene expression. Although these in vivo experiments do not allow to establish whether the stimulation of PRL gene expression is due to an action of the steroid on the pituitary or at the hypothalamic level or alternatively at both sites, it is likely that one of the mechanisms of action of DHEA might be related to a decrease in dopamine release following a depression of TIDA neuron activity. PMID- 8704733 TI - The 24-hour rhythms in plasma growth hormone, prolactin and thyroid stimulating hormone: effect of sleep deprivation. AB - Twenty-four hour secretory rhythms of growth hormone (GH), prolactin (PRL) and thyroid stimulating hormone (TSH) were investigated in 9 normal adult men by means of serial blood sampling at 30 min intervals. The profiles of pituitary hormones were compared in 6 subjects between in normal nocturnal sleep condition and in delayed sleep condition. Plasma GH was measured with use of highly sensitive enzyme immunoassay (EIA) recently developed. Plasma TSH was also evaluated by highly sensitive time-resolved fluorometric immunoassay (TR-FIA). Time series analysis of plasma GH and PRL was performed by auto- and cross- correlation and spectral analysis. The detection limit of EIA for GH was 0.3 pg/ml and all plasma GH levels were within the detectable range of this EIA. Cross-correlation and spectral analysis suggested the presence of approximately 2 3 h rhythmicity of plasma GH. Plasma PRL appeared to have some 24-hour rhythmicity besides its sleep-dependent component. Sleep deprivation caused marked elevation of plasma TSH during night time. It is suggested that there appears two mechanisms regulating GH secretion: one has a sleep-independent and ultradian rhythm and another has a sleep-dependent rhythm. PMID- 8704734 TI - Daily melatonin injections induce cytological changes in pars tuberalis-specific cells similar to short photoperiod. AB - Hypophyseal pars tuberalis (PT)-specific cells are known to exhibit remarkable seasonal changes in morphology especially in photoperiodic animals like the Djungarian hamster Phodopus sungorus. Their high density of melatonin-receptors leads to the supposition that fluctuations in circulating melatonin levels are a crucial factor for the morphological alterations induced by photoperiodic signals. To prove this hypothesis the nocturnal elevation of melatonin in long photoperiods was prolonged by late afternoon administration of melatonin. We investigated whether this treatment induces cytological changes usually observable under short photoperiod. Electron microscopy revealed that in contrast to hamsters maintained in long photoperiods PT-specific cells of hamsters injected with melatonin or those kept in short photoperiods appear inactive, containing a relatively high number of secretory granules, sparse endoplasmatic reticulum, irregularly outlined and invaginated cell nuclei and a high amount of glycogen. Furthermore immunoreactivity for the common alpha-chain of glycoprotein hormones and beta-TSH was significantly weaker in hamsters kept in short photoperiods or daily injected with melatonin than untreated or vehicle injected controls in long photoperiod. These results demonstrate that an exogenous prolongation of the elevated nocturnal melatonin levels causes a similar morphological appearance of PT-specific cells as observed in short photoperiods. It is tempting to speculate that the melatonin signal is a direct 'Zeitgeber' for the transduction of photoperiodic information to the secretory activity in this cell type. PMID- 8704735 TI - Dissociation between behavioral and hormonal responses to the forced swim stress in lactating rats. AB - Retention of immobility in the Porsolt forced swim test is believed to be dependent upon glucocorticoid secretion in male rats. Because lactating females exhibit increased basal glucocorticoid secretion and blunted stress responses, we tested the hypothesis that lactation-induced changes in adrenal glucocorticoid and in circulating estrogen and progesterone levels would improve retention and/or acquisition of immobility. Immobility was recorded during 3 intervals of 5 min on day 1 (acquisition) and one 5 min interval 24 h later (retention). Blood samples were collected before the swim test and at various times after the onset of stress for plasma ACTH and corticosterone (B) determinations. Male rats (young=200 g, old=325 g) were compared to virgin females (V) and to lactating females in early (day 8-10, EL) and late (day 17-19, LL) lactation. Adrenalectomy (ADX) and ovariectomy (OVX) were performed 5 and 10 days prior to testing, respectively. All animals acquired immobility at the end of the 15 min swim on day 1, but only the young male group exhibited a significant retention of immobility on day 2. Total immobility was higher in males than females (V) although basal and stress-induced ACTH and B secretion were comparable on both testing days. Lactational status did not affect immobility in either the acquisition or retention phases. However, stress-induced ACTH secretion was greatly diminished in intact and ADX lactating females (EL and LL) compared to virgins (LL < EL < virgin), demonstrating a clear dissociation between behavioral and neuroendocrine responses. Following ADX, immobility in the retention phase was either decreased in males or increased in lactating females. Finally, OVX decreased immobility in both lactating (EL) and virgin females without significantly altering the magnitude of the ACTH and B responses to stress. In summary, our results demonstrated both sex-related and lactation-related differences in the behavioral and endocrine responses to he forced swim test of Porsolt. Although retention of the immobile response is thought to involve glucocorticoids and/or opioids secreted during the first testing session, we did not find evidence for a direct relationship between basal or stress-induced total corticosterone secretion, the magnitude of ACTH response to stress and behavioral scores in the retention period. However, experimental variables such as body weight, sex and water depth could significantly modify the outcome of behavioral testing and question the validity of glucocorticoid-mediated retention processes. Since the effect of ADX was reversed in lactating females compared to male rats, we hypothesize that glucocorticoid sensitivity of cognitive processes controlling behavioral reactivity is different from that controlling hypothalamic adrenocortical function. Our results also demonstrated a clear dissociation between behavioral and neuroendocrine responses to the swim test, in particular during lactation. In early and late lactation, blunted responsiveness to stress was not caused by enhanced glucocorticoid feedback but might result from modifications in the inhibitory and/or stimulatory inputs to hypothalamic neurons controlling adrenocortical activity. PMID- 8704736 TI - Leukaemia inhibitory factor expression in cultured rat anterior pituitary is regulated by glucocorticoids. AB - Leukaemia inhibitory factor (LIF) is the generally recognized nomenclature for a pleiotropic cytokine that exerts multiple biological effects in different systems. However, its role in the neuroendocrine system is relatively unexplored, with the exception of one report indicating that LIF may act to determine the unique vascular organization of the pituitary gland. In the present study the expression of LIF mRNA has been demonstrated for the first time in the pituitary gland of the rat, in all three pituitary lobes. However, LIF gene expression is restricted to tissues in explant culture where LIF transcripts are readily detectable by Northern Techniques. Similar studies using the reverse transcriptase-polymerase chain reaction (RT-PCR) technique also failed to detect LIF transcripts in fresh tissue from adult animals. These results are consistent with the hypothesis that LIF acts to mediate the response to trauma, a response that may include the induction of neuropeptide expression in the anterior pituitary. In further studies it has also been shown that anterior pituitary LIF mRNA is super-induced in the presence of protein synthesis inhibitors, a response that indicates a role for a labile protein in the attenuation of LIF expression. Both the explantation response, and the response to protein synthesis inhibitory drugs are decreased in the presence of dexamethasone indicating that a glucocorticoid receptor mechanism acts as a general modulatory influence on LIF mRNA levels. These findings are consistent with the presence of multiple regulatory controls on the expression of LIF that serve to restrict its expression to pathological conditions, and indicate that LIF does not participate in normal endocrine physiology. PMID- 8704737 TI - Comparison of the number of vasopressin-producing hypothalamic neurons in rats and humans. AB - The aim of this study was to assess the number and proportion of vasopressin producing neurons in the hypothalamic magnocellular nuclei in rats and humans. Accurate and unbiased neuronal counts were estimated using the optical disector method. Arginine vasopressin-containing neurons were immunohistochemically visualized in formalin-fixed tissue sections. The magnocellular neurons were similar in size and morphology in both species. While the human hypothalamus contained significantly more vasopressin-containing neurons compared with the rat (36-fold increase), the proportion of vasopressin-containing neurons between species was similar. In both species, the majority of supraoptic neurons contained vasopressin, however the proportion of vasopressin-containing neurons in the human paraventricular nucleus was double that of the rat (nearly a 100 fold increase in number). These results suggest that the paraventricular nucleus contributes significantly to the release of vasopressin from the posterior pituitary in humans, whereas in rats vasopressin is mainly released by supraoptic neurons. PMID- 8704738 TI - Evidence that melatonin acts in the pituitary gland through a dopamine independent mechanism to mediate effects of daylength on the secretion of prolactin in the ram. AB - A previous study provided evidence that melatonin acts in the pituitary gland to mediate the effects of daylength on the secretion of prolactin in sheep. This was based on the observation that hypothalamo-pituitary disconnected (HPD) Soay rams showed normal patterns in the changes in the peripheral blood concentrations of prolactin in response to alterations in photoperiod (10-fold higher concentrations under long than short days), and in response to exogenous melatonin (rapid decline following the administration of a constant-release implant of melatonin). The purpose of this study was to establish whether dopamine (DA) might be involved in mediating the effects of melatonin on the secretion of prolactin. Groups of HPD (n = 7) and control Soay rams (n = 8) were treated with vehicle (control, 2.0 ml 0.1 M tartaric acid/saline sc), bromocriptine (DA agonist, 0.06 mg/kg sc) or sulpiride (DA antagonist, 0.6 mg/kg sc), and the acute prolactin responses were measured over the next 4 h. Treatments were carried out under short days (8L: 16D, low prolactin), long days (16L: 8D), high prolactin), and under long days in the presence of a constant release implant of melatonin (low prolactin). The prolactin response to TRH (1.25 micrograms/kg iv) was also measured. Bromocriptine caused a decrease in the plasma concentrations of prolactin in both HPD and control rams under short and long days. Sulpiride had no effect in the HPD rams on any occasion, but caused a very marked increase in the plasma concentrations of prolactin in the control rams under short days, long days, and under long days + melatonin. TRH caused an acute increase in the plasma concentrations of prolactin in the HPD rams under both long and short days although the responses were notably reduced compared with the controls especially under long days + melatonin. Overall, the inhibitory response to the DA agonist in HPD rams indicates the presence of DA D2 receptors linked to functional lactotrophs in the isolated pituitary gland. However, the total lack of a response to the DA antagonist indicates the absence of endogenous DA mechanisms regulating the secretion of prolactin in the HPD rams. The conclusion is that melatonin acts directly on the pituitary gland to mediate effects of photoperiod through a DA-independent mechanism. PMID- 8704739 TI - Effects of chronic octreotide treatment on GH secretory dynamics and tumor growth in rats bearing an ectopic somatotroph (GC) tumor. AB - The effects of octreotide, a long-acting somatostatin agonist selective of the sstr2/sstr3/sstr5 receptor subtypes, on ectopic GH secretion and tumor growth were investigated in Wistar-Furth female rats implanted with GH secreting (GC) cells which express mostly somatostatin receptors of the sstr1 and sstr2 subtypes. Octreotide dose dependently inhibited thymidine incorporation (-57%) and GH secretion (-41%) from GC cells in culture. In vivo, 6 weeks after GC cell implantation, plasma GH, IGF-1 and insulin levels were highly elevated. Cluster analysis of GH secretory dynamics revealed that GH secretion was less pulsatile in GC-implanted than in control animals. Furthermore, in GC-implanted animals, passive immunization either with SRIH or GHRH antisera, did not affect GH plasma levels. Three weeks after GC cell implantation, when tumors became palpable, octreotide (1 micrograms/h/kg BW) or saline was infused constantly for three weeks by osmotic minipumps. In octreotide treated rats, GH, IGF-1 and insulin levels were not different from sham-implanted animals and tumors weight were reduced by 80%. High affinity somatostatin binding sites were found in equivalent amounts on tumors from octreotide-treated or saline-treated animals. These findings indicate that GH secretion in GC-rats is mainly derived from the tumors and independent of hypothalamic control and that octreotide reduces both GH secretion and tumor growth. We conclude that the GC-implanted rat represents a good animal model to test the antisecretory and antitrophic properties of somatostatin analogs in vivo. PMID- 8704741 TI - [How to treat a tumor of the upper excretory tract?]. PMID- 8704742 TI - [How to treat superficial tumors of the bladder?]. PMID- 8704740 TI - A role for the mineralocorticoid receptor in a rapid and transient suppression of hippocampal 5-HT1A receptor mRNA by corticosterone. AB - The interaction between corticosterone and hippocampal 5-HT1A receptor mRNA expression was investigated in male rats. Two days after adrenalectomy, 5-HT1A receptor mRNA was increased in the granule cell layer of the dentate gyrus. Injection of corticosterone led to a significant dose-dependent and transient suppression of 5-HT1A receptor mRNA. The effect of a low dose of corticosterone (50 micrograms/kg) could be blocked by RU 28318, a specific mineralocorticoid receptor antagonist, but not by RU 38486, a glucocorticoid antagonist. The effect of a higher dose of corticosterone (300 micrograms/kg) could be partially blocked by RU 28318, whereas there was no additional effect of RU 38486. These results point to a stringent regulation of hippocampal 5-HT1A receptors by corticosterone that is predominantly mediated by the mineralocorticoid receptor. PMID- 8704743 TI - [How to treat infiltrating cancer of the bladder?]. PMID- 8704744 TI - [How to treat locally advanced and metastatic untreated cancer of the prostate?]. PMID- 8704745 TI - [How to treat hormone-resistant prostatic cancer?]. PMID- 8704746 TI - [How to treat testicular cancer?]. PMID- 8704747 TI - [How to treat hypospadias?]. PMID- 8704748 TI - [How to treat kidney cancer?]. PMID- 8704749 TI - [How to treat ureteral calculi?]. PMID- 8704750 TI - [How to treat interstitial cystitis?]. PMID- 8704752 TI - Reducing patient waiting times through quality assurance methods in La Troncal, Ecuador. AB - Long patient waiting time is a common problem in hospitals and urban health centers in Ecuador and elsewhere. Besides being a leading cause of patient dissatisfaction with health service quality, it is often related to short doctor patient contact times that in turn can seriously reduce the technical quality of care. This article describes a quality improvement effort undertaken by the staff of the La Troncal Health Center in Ecuador, with the assistance of a quality assurance project of the University Research Corporation and the Ecuadorian Ministry of Health. Data on patient waiting times was collected and analyzed, and solutions were devised and implemented using quality assurance techniques. As a result, the average patient's total waiting time fell from 116 minutes per visit to 66, and his or her contact time with health center staff members increased from 11 minutes to 16. It appears that the methods and techniques applied have potential for use elsewhere, because they can be applied easily by health personnel, and their cost of implementation is relatively low. PMID- 8704751 TI - Prescription of antibiotics for mild acute respiratory infections in children. AB - Acute respiratory infections (ARI), the leading class of ailments causing people to seek health care, rarely require antibiotics. Nevertheless, many physicians prescribe them needlessly. Hence, reducing the unnecessary use of antibiotics is one aim of any ARI control program. To help determine whether this aim might be achieved through a combination of refresher training for family physicians and public education campaigns, two 1991 interventions were carried out in four health areas (designated A, B, C, and D) in the city of Havana, Cuba. In each area, 10 clinics staffed by family physicians were selected through simple random sampling. In two areas (A and B), a refresher training program on ARI for health personnel was instituted at each clinic, while in areas A and C a community education program was set up. No intervention was carried out in area D. Simultaneously, from January through December 1991 trained individuals visited and administered a standard questionnaire every 15 days to 1,600 families (40 per clinic) systematically selected by random sampling. The aim of this procedure was to record the number of ARI episodes occurring among children under 5 years old, the treatment chosen in these cases, and whether antibiotics were employed. The results showed that when the two interventions were initiated, antibiotics were prescribed for 26%, 20%, 11%, and 19% of the mild ARI cases occurring in areas A, B, C, and D respectively (P > 0.05). In the period immediately following the interventions, antibiotic prescription rates declined by 26% and 63% in areas A and B, while increasing by 2% and 48% in areas C and D. Overall, prescription of antibiotics in the intervention areas A and B combined decreased by 54% (95% CI: 31-69%). These data suggest that a refresher training program for health personnel can rapidly reduce the unnecessary prescribing of antibiotics for ARI cases, but that public education alone does not appear effective. PMID- 8704753 TI - Breast-feeding and the nutritional status of nursing children in Chile. AB - The work reported here sought to describe the feeding patterns of Chilean children up to 18 months old and their relation to nutritional status. To this end, a survey was conducted in 1993 of 9330 Chilean children under 18 months old who were receiving care through the National Health Service System-which provides care for 75% of all children under age 6. The children, whose mothers or caretakers were interviewed, constituted 94% of a sample selected at random from 102 of the 320 urban health clinics located throughout the country. The interview served to identify the type of feeding (exclusive breast-feeding, breast-feeding plus bottle-feeding, breast-feeding plus solid food, exclusive bottle-feeding, or bottle-feeding plus solid food) and to determine the nutritional status of the participants in terms of standards used by the United States National Center for Health Statistics and the World Health Organization. Children were deemed at risk of malnutrition if they had z scores on the weight-for-age distribution between 1.0 and 2.0 standard deviations below the US/WHO standard and as actually malnourished if they had z scores of over 2.0 standard deviations below the standard. The survey found exclusive breast-feeding prevalences of 86.5%, 66.7%, and 25.3% among infants 1, 3, and 6 months old. Some 12.1% of the participants were found to have a weight-for-age deficiency, 30.7% exhibited a height-for-age deficiency, and 35.7% were found to be over-weight. The prevalence of weight-for age and height-for-age deficiencies were found to be considerably higher among bottle-fed children than among breast-fed children. In general, the results demonstrated the benefits of exclusive breast-feeding through the first 6 months of life, the need to complement exclusive breast-feeding with solid food after that time, and the superior nutritional status of breast-fed children within the age groups studied. PMID- 8704754 TI - Epidemic cholera in Latin America, 1991-1993: implications of case definitions used for public health surveillance. AB - This report presents the various cholera case definitions used by the affected countries of Latin America, shows the numbers of cholera cases and deaths attributable to cholera (as reported by Latin American countries to PAHO through 1993), and describes some regional trends in cholera incidence. The information about how cholera cases were defined was obtained from an October 1993 PAHO questionnaire. In all, 948429 cholera cases were reported to PAHO by affected Latin American countries from January 1991 through December 1993, the highest annual incidences being registered in Peru (1991 and 1992) and Guatemala (1993). The case-fatality rate over the three-year period, and also in 1993, was 0.8%. A general downward trend in the incidence of cholera was observed in most South American countries, while the incidence increased in most Central American countries. A good deal of variation was noted in the definitions used for reporting cholera cases, hospitalized cholera cases, and cholera-attributable deaths. Because of these variations, broad intercountry comparisons (including disease burden calculations and care quality assessments based on case-fatality rates) are difficult to make, and even reported trends within a single country need to be evaluated with care. The situation is likely to be complicated in the future by the arrival of V. cholerae O139 in Latin America, creating a need to distinguish between it and the prevailing O1 strain. For purposes of simplicity, wide acceptance, and broad dissemination of case data, the following definitions are recommended: Confirmed case of O1 cholera: laboratory-confirmed infection with toxigenic V. cholerae O1 in any person who has diarrhea. Confirmed case of O139 cholera: laboratory-confirmed infection with toxigenic V. cholerae O139 in any person who has diarrhea. Clinical case of cholera: acute watery diarrhea in a person over 5 years old who is seeking treatment. Death attributable to cholera: death within one week of the onset of diarrhea in a person with confirmed or clinically defined cholera. Hospitalized patient with cholera:a person who has confirmed or clinically defined cholera and who remains at least 12 hours in a health care facility for treatment of the disease. PMID- 8704755 TI - Influence of PAHO publications on scientific production in the health field in Latin America and the Caribbean. AB - The influence and impact of PAHO publications on scientific production in the field of health in Latin America and the Caribbean was the subject of a study based on a sample of 45 biomedical journals published between 1985 and 1992 in 17 countries of the Region. A total of 8644 works (mostly articles), containing 82,143 citations, were studied. Of these, 3,806 citations were found to refer to works published by PAHO Headquarters in Washington, D.C.-the Boletin de la Oficina Sanitaria Panamericana receiving 1,444 (38% of the total), the English language Bulletin of PAHO receiving 222 (6%), works in PAHO's Scientific Publications Series receiving 1064 (28%), and works in other PAHO publications receiving 1076 (28%). Overall, PAHO publications appeared to account for a significant share of the citations studied. PMID- 8704756 TI - Health and national security. PMID- 8704758 TI - World Health Day 1996: healthy communities. PMID- 8704757 TI - Vaccine self-sufficiency in developing countries. PMID- 8704759 TI - Reflections on hospital quality assurance programs. PMID- 8704760 TI - New, emerging, and re-emerging infectious diseases. PMID- 8704761 TI - An overview of health-related industrial biotechnology in Latin America and the Caribbean. AB - There is some uncertainty about the extent to which Latin America and the Caribbean have participated in the advances of health-related industrial biotechnology. This article reviews the available literature and seeks to provide an overview of the prevailing situation. In general, national governments and multinational agencies have provided most of the health-related biotechnology investments within this region. Efforts to achieve technology transfers, a subject of prime concern, have been developed by a number of programs including the WHO Special Program for Research and Training in Tropical Diseases, the UNDP/UNESCO/UNIDO Regional Biotechnology Program for Latin America and the Caribbean; PAHO's Program for the Regional Development of Biotechnology as Applied to Health; The PAHO/WHO Expanded Program on Immunization (EPI); and PAHO's Regional System of Vaccines (SIREVA). Regarding current production capacity, some successful efforts have been made to produce a variety of therapeutic products including recombinant and natural interferons, interleukins, insulin, and recombinant streptokinase; but in general the region's current potential in this area is at best incipient and uncertain. However, the region does have a limited ability to make diagnostic products and a well-established capacity for vaccine development. Overall, this picture suggests that the region has the potential to play a small but significant role in health-related biotechnology. PMID- 8704762 TI - Public health ... smog clogs urban EDs. PMID- 8704763 TI - Insurance ... health insurance is still far from universal. PMID- 8704764 TI - Policy ... to poor women, Medicaid is a lifesaver. PMID- 8704765 TI - Quality watch ... the Foundation for Accountability. PMID- 8704767 TI - Consumers ... trends on care and coverage. PMID- 8704766 TI - Medicare/Medicaid ... senior citizens are often viewed as resistant to managed care. PMID- 8704768 TI - Quality watch ... how to spot SIDS. PMID- 8704769 TI - Ethics ... when community hospitals merge with Catholic hospitals or networks, reproductive health services often are phased out. PMID- 8704770 TI - Great comebacks: sweet inspiration. PMID- 8704771 TI - Just say grow. Cardinal Health looks beyond drug distribution and sees a healthy future. PMID- 8704772 TI - Merger mania. One isn't the loneliest number. PMID- 8704773 TI - Employer innovation. Wrangler rounds up health care resources. PMID- 8704774 TI - Customer service. Timeliness is next to godliness. PMID- 8704775 TI - Facts & figures. Numbers you can count on. PMID- 8704776 TI - Continuum of care. New ways to treat the old. PMID- 8704777 TI - HospitalPulse ... March 1996. PMID- 8704778 TI - Editorial control: Part 1--A survey of nursing association publications. AB - Editors of association publications may find themselves in conflict with association leaders over the content or philosophy of printed material. To find out how the editors of nursing association publications defined editorial control, this author, herself an editor for an association publication, conducted an informal survey. The results will help editors define their roles, association leaders avoid conflict, and authors identify who makes acceptance decisions for their manuscripts. PMID- 8704779 TI - Editorial control: Part 2--Fostering collegial relationships. AB - Typically the association president and the editor of the association's publication find themselves in discussions over who is ultimately responsible for the publication's style and content. The first article in this set of two focused on the problem, this article describes strategies to use to prevent conflicts between the editor and association leaders. PMID- 8704780 TI - Dictation--the fastest way to write. AB - Nursing leaders are busy making healthcare delivery system changes, so there is little time to write. Yet, the very projects that the nursing leaders are developing would be excellent subjects for articles. Dictation offers the nursing leader an opportunity to write, while taking little time. PMID- 8704781 TI - Functional outcome in the elderly after coronary artery surgery. AB - Patients 80 years of age and older who underwent coronary artery bypass grafting (CABG) were surveyed to determine their postoperative status. Basic activities of daily living were performed by 93%, and 62% of those surveyed reported health satisfaction. CABG can be performed in the elderly population with good functional outcome and an improved quality of life. PMID- 8704782 TI - Preventing allergic reactions to latex. PMID- 8704783 TI - Retooling for the future. PMID- 8704784 TI - The information highway: rules of the road. PMID- 8704786 TI - Nurse advocate: advocate for whom? PMID- 8704785 TI - Medication use in the elderly: a population at risk. PMID- 8704787 TI - Wealth: acquiring it, protecting it, passing it on. PMID- 8704788 TI - The role of the interdisciplinary team in subacute care. PMID- 8704789 TI - Image--is it a problem? PMID- 8704790 TI - Improving care at the end of life. PMID- 8704791 TI - The late effects of poliomyelitis: nursing interventions for a unique patient population. AB - Over half a million Americans who had paralytic poliomyelitis as children or young adults are at risk for experiencing the unexpected late effects of post poliomyelitis syndrome (PPS). When hospitalized or treated for a variety of conditions requiring medical or surgical interventions, these individuals present challenges which include life-threatening complications. PMID- 8704792 TI - Nurses' knowledge, attitudes, and practices in managing urinary incontinence in the acute care setting. AB - Urinary incontinence (UI) is costly, promotes skin breakdown, and delays hospital discharge. While the majority of clinical and research initiatives have focused on managing UI in the nonacute settings, managing UI in the acute care setting is a concern. This review addresses acute care nurses' knowledge, attitudes, and practices regarding UI. PMID- 8704793 TI - Telephone nursing in a general medicine ambulatory clinic. AB - An innovative telephone nursing service was designed to improve care delivery and efficiency in a general medicine clinic. Professional nurses provide general health information and clinical updates and follow intervention guidelines. Patients, physicians, and nurses report high satisfaction and improved patient outcomes. PMID- 8704794 TI - Understanding dysphagia: interventions for care. AB - A significant proportion of patients in both acute and long-term care settings have difficulty in swallowing. These impairments may be temporary, progressive, or permanent in nature. Nurses are in a unique position to assess, coordinate, and manage a treatment regimen. PMID- 8704795 TI - [Reproductive mishaps and infertility in women exposed to DES in utero. A retrospective study of 258 exposed women and 50 unexposed siblings]. PMID- 8704797 TI - [Advantages and disadvantages of consensus conferences]. PMID- 8704796 TI - [Can nitrogen monoxide replace aspirin in the prevention of preeclampsia?]. PMID- 8704799 TI - [Chemotherapy for gynecologic tumors]. PMID- 8704798 TI - [Should nitrogen monoxide replace aspirin in the prevention of preeclampsia?]. PMID- 8704800 TI - [Breast microbiopsies]. PMID- 8704801 TI - [Homage to Marie-Andree Lagroua Weill-Halle]. PMID- 8704802 TI - [Contraception and society]. PMID- 8704803 TI - [Compliance and missed pills in France in 1995]. AB - The analysis of a survey on a representative sample of the whole French women using pill allowed us to measure the number and the consequences of missed pills, to discover the circumstances which induce them and to characterize the most risky groups. It raises the importance of the information given by the doctors. PMID- 8704805 TI - [Evaluation of ovarian sensitivity to gonadotrophins: role of ovarian reserve tests]. AB - Among several parameters involved in the age-dependent reduction in conception rate, intra-ovarian factors, specially the follicular oocyte complex, play a major role. The aim of endocrine challenge tests is to assess ovarian follicular reserve. Basal serum FSH determination on day 3 of the cycle is easily measurable, inexpensive. Its sensitivity is increased with a clomiphene citrate challenge test. Both GnRH agonist stimulation test and exogenous FSH test allow to evaluate more directly the ovarian function. However they are more invasive, expensive and need further standardisation. PMID- 8704804 TI - [Role of conservative therapy and medical treatment in ectopic pregnancy: literature review and clinical trial comparing medical treatment and conservative laparoscopic treatment]. AB - OBJECTIVE: to compare methotrexate (MTX) to laparoscopic salpingotomy for conservative management of ectopic pregnancy (EP). DESIGN: prospective randomized study. PATIENTS: eighty-nine patients were randomized into 2 groups using a random number table. Inclusion criteria were an EP visualized by ultrasound with a pretherapeutic score < or = 13 as assessed by 6 criteria graded from 1 to 3: gestational age, hCG level, P level, abdominal pain, volume of the hemoperitoneum, and diameter of the hematosalpinx. INTERVENTIONS: 1 mg/kg of MTX injected transvaginally into the ectopic pregnancy without anaesthesia or IM administration (1.5 mg/kg) when EP cannot be safely or easily punctured (group 1) versus laparoscopic salpingotomy (group 2). RESULTS: the success rates defined by hCG levels returned to normal (< 10 mlU/mL) were 43 out of 46 in group 1 and 40 out of 43 in group 2. Medical treatment was significantly associated with shorter post-operative stay (24 vs 46 hours). hCG return to normal was quicker after laparoscopic treatment (13 vs 29 days). Reproductive performances were similar in both groups. CONCLUSIONS: in selected cases of EP with a pretherapeutic score < or = 13, MTX treatment appeared to be as safe and efficient as was conservative treatment by laparoscopy, an expectant management should be offered as a treatment option only in women fulfilling the criteria for a good prognostic. PMID- 8704806 TI - [High-dose therapy and hematopoietic cell autotransplantation in the treatment of adult gynecologic tumors]. AB - Autologous bone marrow transplantation for the treatment of gynecologic tumors in adults remains an uncommon therapeutic approach. The feasibility of such high dose therapies is clearly proved, especially with the advent of hematopoietic growth factors and the rescue by the peripheral stem cells to reduce the duration of the chemotherapy-induced myeloid aplasia. The question is to exactly define the place of high-dose therapy in the land of solid tumors. In the treatment of poor prognosis breast cancer, high-dose therapy with autologous bone marrow transplantation or with peripheral stem cells support is able to convert some patients with partial response into complete responders. However, the consequences on overall survival and disease-free survival are not convincing. For metastatic breast cancer and for poor-prognosis tumors (inflammatory breast cancer, axillary metastatic nodes > or = 8), the interest of high-dose therapy has to be determined by randomized studies. These studies are ongoing in USA and in France. For the treatment of poor-prognosis ovarian cancer, the situation is more difficult to appraise. Randomized studies have to be done to precisely define the interest of high-dose therapy in terms of response and disease-free survival for the treatment of ovarian carcinomas. PMID- 8704807 TI - [Pregnancy after breast cancer]. AB - In what ways can a premenopausal woman be pregnant after a breast carcinoma's treatment? A survey organized in 1985 by the French Gynecologic Association gave us the opinion of 316 gynecologists: 50% of them disagreed with this pregnancy because of an accrued risk of relapse. 68 observations were collected including 41 of full term pregnancies; the survival curves were, in each group, similar to those of the controls at the same stage. The interruption doesn't improve the prognosis. This confirms the whole of the published series: a subsequent pregnancy doesn't seem to affect the prognosis of a breast carcinoma; it is important to take the contraception into account so that this pregnancy shall be really desired. PMID- 8704808 TI - [From in vitro fertilization to embryo transfers: evaluation of ten years of reproductive medical assistance in the private sector in France]. AB - The activity of the private-run French AMP centers was analyzed through a retrospective study. The results were given in Toulouse in December 1995. IVF: 50% of activity in France are done in private centers. From 1983 to 1995, 71,974 oocyte retrievals allowing 53,370 embryo transfers were realized and 11,721 pregnancies occurred with 11,088 healthy babies born. ICSI: from 1992 to 1995 3,399 embryo transfers were done. The segmentation rate is 48%. The evolution of the 886 pregnancies thus obtained and the results of the foetal karyotype are detailed in the study. The other assisted reproductive technics (embryo freezing, oocyte donation) are undertaken in the private sector. The results can be found herewith. PMID- 8704810 TI - [Uterine malformations and infertility]. PMID- 8704809 TI - [Ethical problems with the decision of refusal to a request for intra-conjugal in vitro fertilization]. AB - What is our knowledge of the couples who suffer refusal of access to IVF when information is available for only 52% of the pregnancies obtained by IVF? With a retrospective study on IVF refusal datas in a centre where they are available, and an investigation by questionnaire in twelve other centers, the authors tried to evaluate the incidence of these refusal. IVF refusal concern 6 to 30% of the couples addressed, varying on a plurifactorial preselection; but a register including refusal is available in only four of the twelve centers. The main reason for refusal is maternal age. Only three of the twelve centers inquired have a real multidisciplinary staff for access decision. Moreover, if the twelve centers knows the future for the obtained pregnancies, it's true sometimes for only seven centers when IVF is unsuccessful, and never for ten centers when refusal of IVF. Assessment of IVF is incomplete, and improvement is needed based on: a better diffusion of the actual knowledge and its applications; a vast application of "ethical committee procedure" when decision for access is taken; and a global approach of IVF, considering refusal as well as success. PMID- 8704811 TI - [Sexuality after a hysterectomy or much ado about nothing]. PMID- 8704812 TI - [Does hysterectomy modify sexuality?]. PMID- 8704813 TI - [Antibiotics in gynecology. Treatment of acute infections]. PMID- 8704814 TI - [Clinical study of ovulatory cervical mucus]. PMID- 8704815 TI - [History of a rupture]. PMID- 8704816 TI - [Risk of thromboembolism and oral contraceptives]. AB - Four studies recently showed an increased risk of venous thromboembolic events (VTE) in current users of oral contraceptives (OCs). This risk was higher in users of Ocs containing third generation progestagens than in users of Ocs containing levonorgestrel. The excess of VTE attributable to the third generation pills should be about 10 per 100 000 woman-years. Conversely, the risk of death due to cardiovascular disease was not significantly increased in users of third generation pills. In addition, the risk of myocardial infarction associated with these latter types of pill was two-fold lower than the risk associated with the pills containing levonorgestrel, and this decrease was of borderline significance (p = 0.10). The results of these different studies are described and discussed in this paper. PMID- 8704817 TI - [Oocyte fertilizability and embryo development in poor responders]. AB - Oocyte quality (atresia, immaturity), fertilizability, the incidence of abnormal fertilization (parthenogenesis, triploidy) and embryo development (rate of division and morphology) are similar in poor and normal responders. Only, the small number of available embryos (0.9/patient vs 4.4) penalize these patients. PMID- 8704818 TI - [Which stimulation for poor responders? Critical study of results and methods of research]. AB - No therapeutic approach is regularly effective to correct poor ovarian responses. High gonadotropin doses from the beginning of stimulation can increase plasma estradiol level and number of preovulatory follicles, but generally fail to improve ongoing pregnancy rate, in ovulation induction and in vitro fertilization as well. LHRH agonists in long protocol gave in our experience the best results. In short protocol their "flare-up" effect is useful, but plasma LH and progesterone are also increased in follicular phase. The clomiphen citrate--hMG association is more effective on oocyte and embryo quality than quantity. Results of spontaneous cycle and growth hormone or GRF co-treatment are disappointing. Using GnRH agonist micro-doses needs for more precise assessment. Oocyte donation can be the last solution for some of these patients. PMID- 8704819 TI - [Chlamydia trachomatis infection: risk factors]. AB - Genital Chlamydia Trachomatis (CT) infections are the most common cause of tubal infertility. Its prevalence is variable between the authors, but seems to be more frequent in the young population. OBJECTIVE: Evaluate the CT genital infection rate in a population consulting either in a sexually transmitted disease center (STD) or in a family planning unit (FPU). --Judge the interest of a routine screening, and by which technic. --Estimate the information level of population about STD and their prevention. METHODS: 270 men and 331 women were detected by uretral or endocervix sample with an immunofluorescence method (Syva Microtrack). CT serology were also realised. RESULTS: 18.5% of man samples, 20% of woman samples in the STD center and 17.9% of woman samples in the FPU were positive for CT. There is a strong correlation between CT positivity and cervical discharge or cervicitis (RR = 4.64, p < 0.01). Serology was positive in 45.5%, 32.5% and 21.2% respectively for STD center's men, STD center's women and PFU's women. The correlation between samples and serology was 62%. Positive samples were more frequent in the 15-30 years old population and positive serology in the over 30 years old population (RR = 3.57, p < 0.01). There is a correlation between CT positive samples and nulliparity, multiple sexual partner and history of genital infection. CONCLUSION: The high prevalence of CT genital infection observed in our centers justify a routine screening especially in the young population. PMID- 8704820 TI - [Ultrasound-guided biopsies for breast nodules: value of automatic biopsy needle]. AB - Within 18 month, 83 solid breast nodules were biopsied with a long throw biopsy gun with 18 gauge needle (Monopty, Bard Urological, Covington) and a 13 Mhz real time AU 530 (Easote Biomedica, Italy) for the ultrasound guidance. The biopsy procedure was well tolerated with no serious complications, and with no insufficient material. US guided biopsy detected 9 cancers, 47 specific benign lesions (39 fibroadenomas, 1 lipofibroadenoma, 4 sclerosing adenoses, 1 fibromatosis, 1 cyst, 1 lymphadenopathy) and 27 non specific benign fibrocystic dystrophy. In 14 out of the 15 nodules surgically removed, the histological diagnoses were identical. In the one remaining case a fibroadenomas was assessed. In the 36 other cases no evolution occurred. US guided core biopsy with automatic device is probably a very useful approach of uncertain pathological diagnoses in mastology. PMID- 8704821 TI - [Androgens and feminine behavior]. AB - Our actual knowledges of the relationships between androgens and various aspects of the female behaviour are coming from experimental studies performed in various animal species or from observations performed in the woman in situations such as puberty, menstrual cycle, menopause, congenital hyperplasia of the adrenal, polycystics ovaries clearly rising up testosterone levels. The role of androgens in the sexual differentiation, as well as in sexual, aggressive, artistic and cognitive behaviour is investigated in the present work. However, animal data must be modulated in view of specific psychosocial factors in the woman. PMID- 8704822 TI - [Indication of corticotherapy for fetal pulmonary maturation]. AB - Corticosteroid administration before anticipated premature delivery is associated with a large reduction in the incidence of early neonatal death, respiratory distress syndrome, intraventricular hemorrhage and necrotizing enterocolitis. Despite scientific evidence, a minority of women delivering prematurely receive antenatal corticosteroid treatment. Randomized control trials show that there is no short term adverse effects for the infant and mother and no long term adverse effects for the child. We conclude that antenatal corticosteroid use is recommended before premature delivery until 34 weeks gestational age. PMID- 8704823 TI - [Evaluation of the time response of a single dose administration of fenticonazole nitrate]. AB - 174 women with vaginal candidosis confirmed by direct microscopic examination were treated with a single intravaginal fenticonazole nitrate capsule. One third received a second intravaginal capsule three days later. Fenticonazole nitrate cream was also applied daily on the vulvar area in 94.2% of cases. Efficacy was excellent. After one week improvement and complete healing rates were 88% and 80%, respectively. On the second day of treatment pruritus and burning abated in 78% and 85% of women. Mycologic cure demonstrated by microscopic examination was obtained in 92% of cases. Treatment acceptance and tolerance were rated good or excellent by 98% and 92% of patients. Absence of abnormal vaginal discharge due to vaginal capsule was considered as an advantage by women. Results of this trial performed on a large number of patients confirm that product efficacy is fast. PMID- 8704824 TI - Oxygen sensing in the carotid body. AB - In the present article we review in a concise manner the literature on the mechanisms of O2 chemoreception in the carotid body of adult mammals. In the first section we describe the basic structure of the carotid body, and define this organ as a secondary sensory receptor. In the second section is presented the most relevant literature on the O2 metabolism in the carotid body to define the parameters of O2 chemoreception, including hypoxic thresholds and P50 of the hypoxic responses. The final section is devoted to the mechanisms of detection of the hypoxic stimulus. We provide the data in favor and against each of the current three models on O2 chemoreception: the membrane model, the metabolic hypothesis with its different versions and the NAD(P)H oxidase model. PMID- 8704825 TI - Carotid body O2 chemoreception: respiratory and nonrespiratory aspects. AB - The respiratory and nonrespiratory hypotheses of O2 chemoreception in arterial chemoreceptors have been compared to provide a perspective that both mechanisms can potentially coexist. The nonrespiratory membrane mechanism operates at a relatively higher level of PO2 and that respiratory metabolic mechanisms at a lower level. The two mechanisms may overlap in the intermediate range of PO2. PMID- 8704826 TI - Possible role of coupling between glomus cells in carotid body chemoreception. AB - Glomus cells of the carotid body are dye and electrically coupled due to the presence of gap junctions between them. During stimulation by hypoxia, hypercapnia and acidity, about 70% of the cells uncouple to various degrees, whereas the rest either develop tighter coupling or are unaffected. Similar results have been obtained with exogenous administrations of naturally present transmitters such as dopamine and cholinergic substances. Uncoupling is associated with a decrease in junctional conductance and closing of intercellular channels. Tighter coupling is accompanied by opposite effects on these parameters. We think that cell isolation uncoupling leads to release of larger amounts of transmitters toward the carotid nerve sensory terminals. Tighter coupling would reduce the quantities of released chemicals. We may have a delicate titration process modulating the sensory discharge frequency, since a single sensory fiber divides to innervate up to 20 glomus cells. Thus, the discharge frequency of this fiber (the sensory unit) will result from the contributions of its many branches, each impinging on variously active glomus cells. PMID- 8704827 TI - G proteins in carotid body chemoreception. AB - G proteins are signal coupling molecules that play major roles in mediating the effects of transmitters as well as certain sensory signals. In the present study we examined whether oxygen chemoreception in the carotid body is coupled to G proteins. Experiments were performed on carotid bodies isolated from anesthetized cats. Presence of G proteins was examined with ADP-ribosylation of the carotid body membranes. Pertussis toxin (PTX), which inactivates G proteins in neuronal tissues, ADP-ribosylated a single band of carotid body protein with a molecular mass of 41 kDa. With cholera toxin (CTX) only a faint band of protein corresponding to approximately 45 kDa was evident. Perfusing the isolated carotid bodies with PTX (2.5 micrograms/min; 60 min) attenuated the sensory response to hypoxia by 52% of the controls. Perfusion with CTX (50 micrograms/min; for 60 min), on the other hand, increased baseline activity and potentiated the hypoxic response by 125% of controls. Heat-inactivated toxins, however, had no influence on the carotid body sensory response to hypoxia. These results suggest that G proteins are present in the chemoreceptor tissue and they seem to be coupled to the transduction and/or to the transmission of the hypoxic stimulus. PMID- 8704828 TI - Second messenger regulation of tyrosine hydroxylase gene expression in rat carotid body. AB - Previous studies [Czyzyk-Krzeska et al.: J Neurochem 1992;58:1538] demonstrated the relationship between low O2 breathing and tyrosine hydroxylase (TH) gene expression in chemosensory type I cells of the carotid body. In the present study, we have exposed carotid bodies in vitro to hypoxic superfusion media, and subsequently used the reverse transcriptase-polymerase chain reaction technique to measure relative changes in the TH transcript in an effort to elucidate the cellular mechanisms which regulate TH gene expression. Carotid bodies and superior cervical ganglia (SCG) were exposed for 3 h to superfusion media equilibrated with either 10% O2 or 100% O2 and then rapidly frozen on dry ice prior to extraction of total RNA. Hypoxia elevated TH mRNA in the carotid body 3.63 +/- 0.84-fold (mean +/- SEM), while in contrast, these parameters were unchanged in SCG similarly exposed to hypoxic media. Incubation of carotid bodies in zero Ca2+ superfusates greatly attenuated the increase in TH mRNA evoked by hypoxia (1.39 +/- 0.34-fold increase; p < 0.025 compared to normal Ca2+ group). Likewise, exposure to the guanylate cyclase activator, atriopeptin III (100 nM), attenuated the TH mRNA hypoxic response (p < 0.005), while activation of adenylate cyclase with forskolin (10 microM) tended to elevate the response to low O2. Our data suggest that hypoxia, independent of circulating hormones, induces TH gene expression in the carotid body, and that multiple factors, including [Ca2+] and cyclic nucleotides, may be important components of the signal transduction pathway. PMID- 8704829 TI - Carotid body adaptation to hypoxia: cellular and molecular mechanisms in vitro. AB - Chronic hypoxia in vivo promotes long-term changes in the carotid body (CB) response to low PO2. By exposing cultured rat CB chemoreceptors (glomus cells) to 6% O2 for 1-3 weeks, we are investigating the cellular and molecular mechanisms of hypoxic adaptation. Recent studies have uncovered a series of plastic changes in glomus cells including hypertrophy, differential regulation of Na+, Ca2+, and K+ currents, and upregulation of the 'plasticity protein', GAP-43. We have also identified cyclic AMP as a possible intracellular mediator of at least some of these effects of chronic hypoxia. Associated with the changes in ionic currents, glomus cells become electrically more excitable. However, a complete understanding of the physiological response of chronically hypoxic glomus cells to chemostimuli will require more detailed knowledge of the specific alterations in the sensing and signaling pathways, including modifications in neurotransmitter (e.g. catecholamine) functions. PMID- 8704830 TI - Increase in carotid body sensitivity during sustained hypoxia. AB - Ventilation is not constant during steady-state prolonged hypoxia. This raises questions as to the role of the carotid body (CB) in sustained hypoxia. Studies in awake goats using an extracorporeal CB circuit to allow separation of systemic (CNS) and CB blood gases provided evidence that prolonged hypoxia isolated to the CB causes time-dependent increased CB sensitivity to hypoxia. These findings were confirmed in recordings of afferent CB discharge in anesthetized goats. Studies in cats have been compatible with those in goats. It is concluded that prolonged hypoxic stimulation of the CB increases its sensitivity to hypoxia and is at least partly responsible for increased ventilatory drive during prolonged hypoxia such as during altitude acclimatization. PMID- 8704831 TI - The cholinergic hypothesis revisited--an unfinished story. AB - Though exogenously delivered acetylcholine excites the carotid body, past evidence has been considered as unsupportive in assigning acetylcholine an excitatory role during hypoxia or hypercapnia. With ganglionic transmission used as the model, data is presented which aims at blocking the postsynaptic cholinergic receptors, at preventing the presynaptic release of acetylcholine, and at quantitating its release under stimulating conditions. The data support an excitatory role for acetylcholine during hypoxia. PMID- 8704833 TI - Understanding the Medicare coding system. PMID- 8704832 TI - Does catecholamine secretion mediate the hypoxia-induced increase in nerve activity? AB - Catecholamine secretion from carotid body glomus cells is hypothesized to cause the hypoxia-induced increase in nerve activity. To test aspects of this hypothesis, tissue catecholamine and single-fiber nerve activity was measured from rat carotid bodies in vitro. Hypoxia (1-min duration, 0 Torr at nadir) caused a rapid increase in catecholamine release and nerve activity, consistent with the hypothesis, but repetitive hypoxias interspersed with short rest periods resulted in a much greater decline in catecholamine release than nerve activity. Furthermore, pretreatment with reserpine (24 h) nearly abolished catecholamine release, but nerve response was not different than untreated controls. These results suggest that catecholamine secretion is not causal to the increase in nerve activity of rat carotid body. PMID- 8704834 TI - Selection of appropriate support surfaces. AB - Many claims are made regarding the appropriate selection of support surfaces. ET nurses do not need to "reinvent the wheel" when they decide which system is best for their patients, but they do need to be aware of the most current scientific and expert recommendations. The WOCN and the Agency for Health Care Policy and Research have published guidelines that list concrete suggestions about the prediction and prevention of pressure ulcers in adults. I developed an algorithm that incorporates the two groups' recommendations and the most recent scientific research. This algorithm can be used in multiple nursing settings. PMID- 8704835 TI - A study of risk factors in patients placed on specialty beds. AB - Air-fluidized and low-air loss beds have become common throughout the United States. Many studies have demonstrated the efficacy of these beds in the prevention and treatment of pressure ulcers. Determining which patients truly need this therapy and which could have positive outcomes with other pressure relieving or pressure-reducing devices can be difficult. The purpose of this research was to examine medical diagnoses, total Braden Scale scores, Braden Scale subscores (sensory perception, moisture, activity, mobility, nutrition, and friction/shear), nutritional levels, and comorbid conditions among patients placed on specialty beds for prevention and treatment of pressure ulcers. A retrospective chart review of 47 patients who used specialty beds during an 18 month period was used to determine factors that were clinically significant. PMID- 8704836 TI - Selection of a standard hospital mattress: data-based decision making. AB - By means of a scientific method, standard hospital mattresses were evaluated for pressure-relieving properties and patient comfort. Vendors' written materials and product demonstrations led to the initial selection of seven mattresses. On the basis of findings from a survey of staff nurses and environmental services and pressure readings obtained with three healthy volunteers, three mattresses were selected for intensive evaluation. Eighteen mattresses (six of each type) were placed in six clinical care units; at 2-week intervals, interface pressure readings (occipital, sacral, and heel) were performed on patients by means of a computerized measurement system. Caregiver and patient questionnaires (n = 100) were analyzed for clinical significance and patients' responses. We conclude that there are no significant differences among the three mattresses tested in pressure-reducing capabilities, nursing functions, or patient comfort. PMID- 8704837 TI - Evaluation of three types of support surfaces for preventing pressure ulcers in patients in a surgical intensive care unit. AB - Because critical care nurses recognize that many of their patients are at risk for pressure ulcer development, they provide them with support surfaces that can reduce this risk. Few reported studies, however, are available to help these nurses choose these surfaces wisely. This project was a new-product evaluation that compared the clinical effectiveness of three types of support surfaces: two dynamic mattress replacement surfaces and a static foam mattress replacement. Members of a convenience sample of 110 patients admitted to a surgical intensive care unit each used one of the three support surfaces. When each patient was placed on one of the three surfaces, the evaluators rated likelihood of pressure ulcer development (Braden Scale score) and assessed the skin for pressure ulcers. The evaluators repeated the Braden Scale score weekly and the skin assessment three times each week. Nine patients (8%), three patients on each support surface, acquired pressure ulcers. The log-rank test did not find a statistically significant difference between the three types of support surfaces with respect to the risk of pressure ulcer development. Stepwise Cox proportional hazards regression revealed a statistically significant relationship between the risk of developing a pressure ulcer, the averaged total Braden Scale score, and the averaged score for the sensory perception subscale of the Braden Scale. Although these three surfaces were comparable in effectiveness, they were not comparable in cost. Both dynamic mattress replacement surfaces cost approximately $2000 each, whereas the cost of the static foam mattress replacement was only $240 each. The results of this product evaluation should encourage other nurses to evaluate patient care products carefully before making recommendations. PMID- 8704838 TI - Trends in pediatric ostomy surgery: intestinal diversion for necrotizing enterocolitis and biliary diversion for biliary hypoplasia syndromes. AB - Ostomies are placed in children for different indications than in the older population. Many ostomies of childhood are placed because of congenital or neonatal problems that require temporary or long-term diversion to stabilize the neonatal patient. Necrotizing enterocolitis, the most common reason for placement of neonatal colostomies and ileostomies, is increasing in frequency as more prematurely born infants survive. Recently, there has been an increase in treatment of various biliary hypoplasia syndromes with biliary cutaneous diversion. Children with biliary hypoplasia syndromes are a challenging group of patients who frequently can be helped by ostomies. This article reviews current information on biliary cutaneous diversion for the biliary hypoplasia syndromes and intestinal diversion for necrotizing enterocolitis. PMID- 8704839 TI - Urinary catheter-related infections among home care patients. AB - Urinary catheterization is a common indication for home care nursing services among older adults, and catheter-related urinary tract infections commonly occur among these patients. The purpose of this study was to determine the rate of urinary catheter-related infections among a group of elderly adults cared for in the home, the characteristics of those who acquire urinary tract infections as opposed to those who do not become infected, and the influence of the interval between catheter changes on the incidence of urinary tract infections. PMID- 8704840 TI - "I will manage": promoting continence through community education. AB - Urinary incontinence affects approximately 10 million Americans, mostly elderly persons in community and institutional settings. Despite the prevalence of UI and an estimated annual total cost of $ 10 billion in the United States alone, most affected persons do not seek help for incontinence. This is chiefly because of embarrassment or because they are not aware that help is available. In this article, we describe a community education and support program for persons with fecal and urinary incontinence. The program is based on the "I Will Manage" model from the Simon Foundation for Continence, founded to "increase public awareness of incontinence, remove social stigma attached to this disability, and provide education to those suffering from incontinence, their families, and the professionals responsible for their care." The program provides a comprehensive and practical framework to promote continence through community education. In this article, we outline the program and provide information we gathered from hosting it. PMID- 8704841 TI - Infected graft in a popliteal site. PMID- 8704842 TI - The rewards of mentorship. PMID- 8704843 TI - Caring for caregivers: home care nursing's challenge. AB - Home care nurses frequently manage patients who also receive a substantial amount of care from a lay caregiver in the home. Because many of these care givers are themselves frail and elderly, the home care nurse must assist both the patient and the caregiver to cope with demands for physical care, complex technology, and behavioral manifestations of chronic conditions. Specialty practice nurses are in an ideal position to assist both the primary visiting nurse and the lay caregiver in the management of complex wound, ostomy, and continence care associated with early discharge from the hospital. PMID- 8704844 TI - Pressure ulcer incidence in an acute care setting. AB - The purpose of this prospective study was to determine the incidence of pressure ulcers and to examine factors related to pressure ulcer development in patients in an acute care setting. Adult medical and surgical patients who were free of pressure ulcers at admission were assessed within 36 hours of admission and then three times per week for 2 weeks or until discharge. Instruments included a demographic data form, a skin assessment form, and the Braden Scale for Predicting Pressure Sore Risk. Most subjects had 46 assessments completed. The sample consisted of 149 subjects, with a pressure ulcer incidence rate of 13.4% (n = 20). Subjects who acquired pressure ulcers had lower hemoglobin levels (t = 2.17, p = 0.03), spent more time in bed (t = 3.90, p = 0.0001), and spent less time in a chair (t = 3.2, p = 0.002) than those who did not acquire pressure ulcers. A stepwise logistic regression analysis was used to calculate risk of pressure ulcer development. In the final model, hemoglobin level and hours spent in bed continued to be predictors of pressure ulcer development (chi 2 = 9.306, df = 2, p = 0.0095). All 20 subjects who acquired pressure ulcers were further categorized into groups with stage I (n = 12) or stage II (n = 8) ulcers. Patients with stage I pressure ulcers were primarily receiving post-surgical care (67%), whereas patients who acquired stage II ulcers had medical conditions that affected tissue perfusion, such as respiratory diseases (50%) and diabetes mellitus (12%). PMID- 8704845 TI - The value of absorbent products and containment devices in the management of urinary incontinence. PMID- 8704846 TI - Medicaid claims history of Florida long-term care facility residents hospitalized for pressure ulcers. AB - The purpose of this study was to identify patterns of admission, discharge, and readmission between hospital and long-term care facility among a group of Florida long-term care facility residents with pressure ulcers whose care was paid for by Medicaid. A patient-specific, longitudinal claims history database was constructed from data provided by the Florida Department of Health and Rehabilitative Services. This database was used to determine and analyze hospital admissions for pressure ulcer care among Medicaid recipients cared for in a long term care facility. Analysis of the data determined that more than half of the Medicaid-covered long-term care facility residents who formed the target study group (54.57%) had multiple hospital admissions associated with pressure ulcers. Pressure ulcer hospital admissions amounted to a program cost of $9.9 million. PMID- 8704847 TI - Changes in Medicare reimbursement for ostomy supplies: an overview. AB - Medicare reimbursement and utilization for ostomy supplies have undergone dramatic change in the last year. This article provides an introduction for health care professionals to the Medicare system, Durable Medical Equipment Regional Carriers, Medicare participating and nonparticipating suppliers, and practical examples of how reimbursement may affect the selection of ostomy products. The topics covered refer to Medicare Part B. ET nurses must be cognizant of the ongoing changes in reimbursement and must collaborate with suppliers, manufacturers, and other health care professionals to provide patients with the highest quality care, products, and services. PMID- 8704848 TI - Sexual issues for the person with an ostomy. AB - The person with an ostomy faces many adjustments that affect sexuality. He or she must adjust to changes in body image and in sexual self-image, all while learning new techniques of self-care. The PLISSIT (Permission, Limited Information, Specific Suggestion, and Intensive Therapy) theory of Annon is useful when providing sexual health care education. Communication, education, and experimentation are the key elements to sexual rehabilitation for the person with an ostomy. Multiple holistic therapeutic suggestions are proposed for client education. PMID- 8704849 TI - Mindfulness and people with stomas. AB - Persons with a stoma must contend not only with the immediate physical changes produced by the operation but also with psychologic adjustments, changes in body image, changes in self-concept, and the chronic stress of stoma care and physical functioning. Mindfulness training can enable these persons to make the necessary adjustments in a more realized and conscious manner. The practice of mindfulness training has been demonstrated to be effective in dealing with clinical and psychologic problems. Mindfulness is defined as being fully awake and is characterized as being nonjudging, having a beginner's mind, being trusting, nonstriving, accepting, and letting go. Mindfulness training is described, and roadblocks to the practice of mindfulness are discussed and exemplified. Resources on the practice of mindfulness and applications to persons with ostomies are provided. Mindfulness training is proposed as an adjunct to patient education or support groups. PMID- 8704850 TI - A physiologic approach to bowel training. AB - Bowel training is an effective management option for many patients with dysfunctional bowel elimination patterns and neurogenic fecal incontinence. To be effective, a bowel training program must be based on sound physiologic principles and must be individualized for each patient. This article includes a review of the structures and physiologic processes controlling normal defecation and the physiologic principles governing bowel training. The steps involved in successful implementation of a bowel training program are discussed, and case studies are used to illustrate the principles. PMID- 8704851 TI - Standardized ET nursing database: imagine the possibilities. AB - A national database of the components of ET nursing practice currently does not currently exist. Justification for ET nurse services requires hard data that demonstrate quality and cost-effectiveness of care. Lacking hard data, the value of the services provided by ET nurses remains difficult to quantify. The Nursing Minimum Data Set is the first step in building a nursing database. The purpose of this article is to familiarize ET nurses with the concepts of nursing information systems, the Nursing Minimum Data Set, and classification systems. Implications for ET nursing practice are also presented. PMID- 8704852 TI - Leg ulcers with exposed bone and tendon. PMID- 8704853 TI - Clean versus sterile technique. PMID- 8704855 TI - Exposure to crocidolite and the incidence of different histological types of lung cancer. AB - OBJECTIVES: To estimate the relations between exposure to both tobacco smoke and crocidolite and the incidence of various histological types of lung cancer. METHODS: In 1979 all former workers from the Wittenoom asbestos industry who could be traced were sent a questionnaire on smoking history. Of 2928 questionnaires sent, satisfactory replies were received from 2400 men and 149 women. Of the men, 80% had smoked at some time and 50% still smoked. Occupational exposure to crocidolite was known from employment records and follow up was maintained through death and cancer registries in Australia with histological diagnoses obtained from the relevant State Cancer Registry. Conditional logistic regression was used to estimate the effects of tobacco and asbestos exposure on incidence of different cell types of lung cancer in a nested case-control design. RESULTS: Between 1979 and 1990, 71 cases of lung cancer occurred among men in this cohort: 27% squamous cell carcinoma, 31% adenocarcinoma, 18% small cell carcinoma, 11% large cell carcinoma, and 13% unclassified or indeterminate. Two of the classified cases and one unclassified case had never smoked. The incidence of both squamous and adenocarcinoma types of lung cancer were greatest in ex smokers and in those subjects with the highest levels of exposure to crocidolite. After adjustment for smoking habit, the increase in incidence of lung cancer with increasing exposure to crocidolite was greater for squamous cell carcinoma than for adenocarcinoma. CONCLUSIONS: The results from this study have shown significant exposure-response effects for exposure to crocidolite, and both adenocarcinoma and squamous cell carcinoma of the lung. They also provide some further evidence against the theory that parenchymal fibrosis induced by asbestos is a necessary precursor to asbestos induced lung cancer. PMID- 8704856 TI - Compensating lung cancer patients occupationally exposed to coal tar pitch volatiles. AB - OBJECTIVES: To investigate the process of deciding on compensation claims by lung cancer patients exposed occupationally to coal tar pitch volatiles. METHODS: For each case of lung cancer the probability that it was caused (probability of causation (PC)) by coal tar pitch volatiles was expressed as an increasing function of cumulative exposure to benzo-a-pyrene-years. This was assessed from several exposure-response models fitted to data from a large epidemiological study of aluminum production workers. For some models, PC depended also on the smoking habit of the cancer patient. RESULTS: Estimation of relative risk by exposure group indicated that over 50% of lung cancers were attributable to coal tar pitch volatiles (PC > 50%) at exposures above 100 micrograms/m3-years benzo(a)pyrene. A linear relative risk model indicated that 50% PC was first achieved at 342.2 micrograms/m3-years benzo(a)pyrene, or 190.1 micrograms/m3 years benzo(a)pyrene according to the upper 95% confidence limit for risk increment. Corresponding figures for a power curve model were 210.3 and 45.9. With these five figures as compensation criteria compensation would have resulted in 31.4%, 2.7%, 19.2%, 15.7%, and 39.2% of cancers studied, compared with an estimated total proportion of cancers studied attributable to coal tar pitch volatiles of 15%-26%. If risks due to coal tar pitch volatiles and smoking multiply, PC does not depend on the amount smoked. If the two risks are additive, however, PC depends on the amount smoked according to a formula, with the figures mentioned applying to an average smoking history (24.4 pack-years). CONCLUSION: Because of its simplicity and because it falls within the range of criteria based on several more sophisticated approaches, we prefer the criterion of 100 micrograms/m3-years, based on the relative risks by exposure group. However, the compensation board of the Canadian province of Quebec, on consideration of these alternatives, has proposed as a criterion that the upper 95% confidence limit of PC for the patient be at least 50%, assuming an additive relative risk model and allowing for their smoking habit. PMID- 8704854 TI - Occupational risk factors for female breast cancer: a review. AB - OBJECTIVES: Although progress has been made in identifying personal risk factors and in improving treatment for female breast cancer, incidence rates continue to increase. With women now occupying a sizable fraction of the workforce, it is worth inquiring whether there are occupational risk factors for breast cancer. This is a review of occupational studies on female breast cancer. METHODS: Suitable reports and published articles with associations of female breast cancer and occupation were identified from technical reports, by searching the MEDLINE bibliographic data base, and by reviewing each paper on cancer that was published in 20 major journals during the period from about 1971-94. RESULTS: A total of 115 studies were identified; 19 studies relied exclusively on data collected for administrative purposes, and there were four incident case-control studies and 92 cohort studies. Although data for individual industries, occupations, and exposures were sparse, there was limited evidence of an association with employment in the pharmaceutical industry and among cosmetologists and beauticians. Associations were also found for chemists and occupations with possible exposure to extremely low frequency electromagnetic fields, but potential methodological weaknesses preclude drawing any definite conclusions. There was little support for increased risks among textiles workers, dry cleaning workers, and nuclear industry workers. CONCLUSIONS: Few high quality occupational studies directed specifically toward women have been carried out to allow the unambiguous identification of occupational risk factors for breast cancer. It is suggested that investigations that account for non-occupational risk factors and that assess exposure in a more detailed way be carried out. One strategy already suggested is to conduct population based, case-control studies in which subjects are interviewed about their occupational histories and exposure to chemical and physical agents which are then attributed from the job descriptions by a team of experts. These studies can then be supplemented when necessary with cohort studies of specific populations. PMID- 8704857 TI - Melanoma and occupation: results of a case-control study. AB - OBJECTIVES: Associations between occupational exposures and the occurrence of cutaneous melanoma were examined as part of a large population based case-control study of 19 cancer sites. METHODS: Cases were men aged 35 to 70 years old, resident in Montreal, Canada, with a new histologically confirmed cutaneous melanoma (n = 103). There were two control groups, a randomly selected population control group (n = 533), and a cancer control group (n = 533) randomly selected from among subjects with other types of cancer in the large study. Odds ratios for the occurrence of melanoma were calculated for each exposure circumstance for which there were more than four exposed cases (85 substances, 13 occupations, and 20 industries) adjusting for age, ethnicity, and number of years of schooling. RESULTS: Significantly increased risk of melanoma was found for exposure to four substances (fabric dust, plastic dust, trichloroethylene, and a group containing paints used on surfaces other than metal and varnishes used on surfaces other than wood), three occupations (warehouse clerks, salesmen, and miners and quarrymen), and two industries (clothing and non-metallic mineral products). CONCLUSIONS: Most of the occupational circumstances examined were not associated with melanoma, nor is there any strong evidence from previous research that any of those are risk factors. For the few occupational circumstances which were associated in our data with melanoma, the statistical evidence was weak, and there is little or no supporting evidence in the scientific literature. On the whole, there is no persuasive evidence of occupational risk factors for melanoma, but the studies have been too small or have involved too much misclassification of exposure for this conclusion to be definitive. PMID- 8704858 TI - Investigation on neurotoxicity of occupational exposure to cyclohexane: a neurophysiological study. AB - OBJECTIVES: To examine the effect of occupational exposure to cyclohexane on the peripheral nervous system. METHODS: A nerve conduction study was performed on 18 workers exposed to cyclohexane in a luggage factory and on age and sex matched occupationally unexposed controls. 12 workers had been exposed to n-hexane (median 2.8 years) before the start of exposure to cyclohexane. To confirm the effect of exposure, a follow up study was performed on nine workers one year after the first study. The mean exposure to cyclohexane was 1.2 years in the first study. A symptom survey was performed. The exposure was measured by air sampling of the breathing zone of each worker. The urinary metabolite cyclohexanol was also monitored. RESULTS: The concentration of airborne cyclohexane ranged from 5 to 211 ppm. The urinary concentration of cyclohexanol ranged from 0.12 to 1.51 mg/l. There was a strong correlation between the cyclohexane exposure in personal air and urinary cyclohexanol. No differences were found in nerve conduction velocities (NCV) between workers exposed to cyclohexane and age and sex matched controls. The results of the follow up study showed significant improvements in peroneal motor NCV (P < 0.01) and sural sensory NCV (P < 0.05) and in ulnar motor distal latency (MDL, P < 0.05) and peroneal MDL (P < 0.05) compared with the first study. Although the past n-hexane exposure affected the first neurophysiological study, the effect had disappeared in the second study, one year later. CONCLUSION: Occupational exposure to the concentrations of cyclohexane experienced in this study had no adverse effects on the peripheral nervous system. PMID- 8704859 TI - Biomarkers of exposure, antibodies, and respiratory symptoms in workers heating polyurethane glue. AB - OBJECTIVES: The pathogenic basis of respiratory disorders associated with isocyanates are still obscure. One reason for this is the lack of good estimates of human exposure. In this study exposure was estimated by measurement of isocyanate metabolites in biological samples. METHODS: In a factory using isocyanate based polyurethane (PUR) glue, isocyanate concentrations in air were measured by liquid chromatography. Samples from 174 employees were analysed for metabolites of 4,4'-methylene diphenyl diisocyanate (MDI) in plasma (P-MDX) and urine (U-MDX). After hydrolysis, 4,4'-methylenedianiline was measured by gas chromatography-mass spectrometry (GC-MS). The employees were screened for work related respiratory symptoms and tested for specific immunoglobulin E (IgE) and IgG antibodies directed against isocyanate conjugated to human serum albumin. RESULTS: The time weighted isocyanate concentrations in air were low (MDI < 0.2 7; hexamethylene diisocyanate (HDI) < 0.1-0.7; 2,6-toluene diisocyanate (TDI) < 0.1 microgram/m3). All subjects had detectable P-MDX and U-MDX. There were significant associations between the estimates of exposure to thermal degradation products of an MDI based glue and P-MDX (range < or = 0.10-5.5 micrograms/l); and U-MDX (< or = 0.04-5.0 micrograms/g creatinine); in cases of heavy exposure. P MDX and U-MDX were associated with each other (r = 0.64; P = 0.0001), work related symptoms (P-MDX: P = 0.03; Mann-Whitney U test), and serum concentrations of MDI specific IgG antibodies (r = 0.26; P = 0.0007). Unexpectedly, high P-MDX and U-MDX concentrations were also encountered in workers cutting textile (P-MDX 2.4-4.5 micrograms/l; U-MDX 0.81-3.8 micrograms/g creatinine); the reason is still unknown. Equally unexpected, there were significant negative associations between P-MDX and liver function tests. CONCLUSIONS: The results clearly show the value of biomarkers for isocyanate exposure; in particular, P-MDX is useful. Further, these results show the risk connected with thermal degradation of PUR. PMID- 8704860 TI - Epidemiological study of hard metal asthma. AB - OBJECTIVES: To elucidate factors contributing to hard metal asthma, the entire workforce of a corporation producing hard metal tools (n = 703) was examined. METHODS: The variables evaluated were the atopy reflected by immunoglobulin E (IgE) antibody against mite allergen, history of exposure to hard metal, smoking, concentration of airborne cobalt, specific IgE antibody reaction against cobalt, and the respiratory symptom of attacks of reversible dyspnoea with wheeze (asthmatic symptoms). RESULTS: Univariate analysis showed that the prevalence of the asthmatic symptoms was significantly higher in formerly and currently exposed male workers than in non-exposed male workers. Positive IgE reaction against cobalt was found in seven men (2.0%), all of whom had asthmatic symptoms. Furthermore, it was found that atopy, positive IgE antibody against cobalt, and age of 40 or older were significantly correlated with asthmatic symptoms. Multilogistic analysis on the same factors and smoking in all of the workers showed that the age, experience of hard metal exposure, and atopy were significant risk factors associated with the asthmatic symptoms. Multilogistic analysis of data for currently exposed and non-exposed workers also showed that age and atopy were risk factors, and that the exposure to cobalt at the low concentration (at or below 50 micrograms/m3) but not at the higher concentration was a significant risk factor. Exposure to mist of coolants containing ionic cobalt, used during grinding, was not found to be any more hazardous in terms of onset of asthmatic symptoms than exposure to hard metal dust containing metallic cobalt. CONCLUSIONS: Accordingly, it is concluded that both environmental factors and individual susceptibility should be taken into consideration in efforts to reduce the prevalence of hard metal asthma. PMID- 8704861 TI - Decreased ventilatory function in hard metal workers. AB - OBJECTIVES: To study individual effects on pulmonary function of exposure to hard metal including cobalt. METHODS: All of the workers in a hard metal company (583 men and 120 women) were examined for smoking, respiratory symptoms, ventilatory function, occupational history of exposure to hard metal, and present exposure to airborne cobalt. The ventilatory function indices (forced vital capacity (FVC), forced expiratory volume in one second (FEV1), forced expiratory volume in one second per cent (FEV1%), peak expiratory flow (PEF), mid-maximal flow (MMF), forced expiratory flow at 50% vital capacity (V50), forced expiratory flow at 25% vital capacity (V25)) were standardised for height and age and expressed as a percentage of predicted values. RESULTS: Two way analysis of variance of indices of ventilatory function showed that an interaction of hard metal exposure and smoking decreased %V50 for both men and women. Among the currently exposed men, those with asthmatic symptoms (defined as reversible dispnoea with wheeze) had significantly lower %FVC, %FEV1%, %PEF, %MMF, %V50, and %V25 than did workers without asthma. The ventilatory disfunction did not differ between exposed and non-exposed workers with asthmatic symptoms. Even among the men without asthmatic symptoms, %V50 was significantly lowered by the interaction of hard metal exposure and smoking. The multilinear regression analysis of indices of ventilatory function for all of the subjects on sex, smoking (Brinkman index), exposure to hard metal, and asthmatic symptoms showed that asthmatic symptoms and smoking had significant effects on all variables and that the decrease in %V25 was associated with hard metal exposure. In the currently exposed and non-exposed workers, multilinear regression analysis applying indices for cobalt exposure (mean cobalt concentration, duration of exposure, and cumulative dose) showed that not only asthmatic symptoms or smoking but also duration of exposure had significant decreasing effects on %FVC, %MMF, and %V25. CONCLUSIONS: Occupational exposure to hard metal probably causes impairment of ventilatory function in a dose dependent manner. PMID- 8704862 TI - Distribution and determinants of personal exposure to nitrogen dioxide in school children. AB - OBJECTIVES: To assess the distribution of personal exposures to nitrogen dioxide (NO2) in school children, and to investigate factors that might influence personal exposure. METHODS: NO2 exposures were assessed by use of passive diffusion tubes for 46 children aged 9-11 years, selected from two Southampton schools. The tubes were worn for seven days, and parallel measurements were made with static samplers in the child's kitchen, living room, classroom, and playground. Information about potential exposures was collected by questionnaire. RESULTS: Personal exposures to NO2, averaged over seven days, ranged from 11 to 257 micrograms/m3 (6 to 137 ppb) with a geometric mean of 36 micrograms/m3 (19 ppb). Exposures correlated with concentrations of NO2 recorded in the home, but the relation was far from exact. Factors associated with increased personal exposure included the use of gas appliances in the home, living with one or more smokers, and travel to school by means other than a car. However, together these variables only explained a small part of the variation in personal exposures. CONCLUSIONS: These findings reinforce the need for personal monitoring of exposure in studies investigating potential health effects of NO2 in children. PMID- 8704863 TI - Prevalence of the sick building syndrome symptoms in office workers before and after being exposed to a building with an improved ventilation system. AB - OBJECTIVE: To find if the prevalence of symptoms associated with sick building syndrome decreased among office workers after moving to a building with improved ventilation (after controlling for potential confounders). METHODS: Workers in five buildings in 1991 all moved in 1992 into a single building with improved design, operation, and maintenance of the ventilation system. All buildings had sealed windows with mechanical ventilation, air conditioning, and humidification. Workers completed a self administered questionnaire during normal working hours in February 1991 and February 1992. The questionnaire encompassed symptoms of the eyes, nose and throat, respiratory system, skin, fatigue, headache, and difficulty concentrating, personal, psychosocial, and work related factors. During normal office hours of the same week environmental variables were measured. RESULTS: The study population comprised 1390 workers in 1991 and 1371 workers in 1992 who represented more than 80% of the eligible population. The prevalence of most symptoms decreased when workers moved to the new building: skin (54%), respiratory system (53%), nose and throat (46%), fatigue (44%), headache (37%), eyes (23%). These findings were all significant and remained generally similar after controlling for personal, psychosocial, and work related factors. Furthermore, more than 60% of workers symptomatic in 1991 were asymptomatic in 1992 for all types of symptoms. In contrast, less than 15% of workers were asymptomatic in 1991 but symptomatic in 1992 for all types of symptoms. CONCLUSION: In this study, the prevalence of most symptoms usually associated with the sick building syndrome decreased by 40% to 50% after workers were transferred to a building with an improved ventilation system. The results show that it is possible to diminish the prevalence of symptoms associated with the sick building syndrome among office workers occupying a building with mechanical ventilation, air conditioning, and sealed windows. PMID- 8704864 TI - Glutathione transferase activity and formation of macromolecular adducts in two cases of acute methyl bromide poisoning. AB - OBJECTIVES: To determine the activity of glutathione transferase and to measure the S-methylcysteine adducts in blood proteins, after acute inhalation exposure to methyl bromide. To examine the influence of the polymorphism of glutathione-S transferase theta (GSTT1) on the neurotoxicity of methyl bromide. METHODS: Two workers acutely exposed to methyl bromide with inadequate respiratory protective devices were poisoned. Seven weeks after the accident, blood samples were drawn from both patients, for measurement of glutathione transferase activity in erythrocytes (conjugator status--that is, GSTT1 phenotype) and measurement of binding products of methyl bromide with blood proteins. Conjugator status was determined by a standard procedure. The binding product of methyl bromide, S methylcysteine, was measured in globin and albumin. RESULTS: Duration and intensity of exposure were identical for both patients as they worked together with the same protective devices and with similar physical effort. However, one patient had very severe poisoning, whereas the other only developed mild neurotoxic symptoms. The first patient was a "conjugator" with normal glutathone transferase activity, whereas this activity was undetectable in the erythrocytes of the second patient, who consequently had higher concentrations of S methylcysteine adduct in albumin (149 v 91 nmol/g protein) and in globin (77 v 30 nmol/g protein). CONCLUSIONS: Methyl bromide is genotoxic and neurotoxic. Its genotoxicity seems to be the consequence of the alkylating activity of the parent compound, and conjugation to glutathione has a protective effect. The data presented here suggest a different mechanism for methyl bromide neurotoxicity which could be related to the transformation of methylglutathione into toxic metabolites such as methanethiol and formaldehyde. If such metabolites are the ultimate toxic species, N-acetylcysteine treatment could have a toxifying rather than a detoxifying effect. PMID- 8704865 TI - A bibliometric study of the trend in articles related to epidemiology published in occupational health journals. AB - OBJECTIVES: To study the role assumed by epidemiology in occupational health literature and characterise its change over the years. METHODS: A bibliometric study was conducted with a MEDLINE search to evaluate 9024 articles published in eight representative occupational health journals for the period 1980-93. Articles related to epidemiology were found by descriptors indexed in the articles and their numbers and proportions among all published articles was tabulated for the study period. RESULTS: The proportion of source items indexed by epidemiology as a descriptor increased over threefold from 7.9% (42/534) in 1980 to over 25% after 1990. Epidemiology was indexed most frequently as a subheading associated with other descriptors-for example, occupational diseases epidemiology. Cohort studies had the largest increase in the descriptors that indicated epidemiological type of studies during the period studied. CONCLUSION: Epidemiology is assuming increasing importance in occupational health literature. This was shown quantitatively and qualitatively by the use of descriptors in the MEDLINE database. PMID- 8704866 TI - Mutation rates at the glycophorin A and HPRT loci in uranium miners exposed to radon progeny. AB - OBJECTIVES: To find whether a relation exists between estimated levels of exposure to radon and its progeny and mutations in hypoxanthine phosphoribosyl transferase (HPRT) and glycophorin A in a cohort of former uranium miners. METHODS: A cohort study involving a sample of miners from the Radium Hill uranium mine in South Australia, which operated from 1952 to 1961. Radiation exposures underground at Radium Hill were estimated from historical radon gas measures with a job exposure matrix. Workers from the mine who worked exclusively above ground according to mine records were selected as controls. In 1991-2 miners were interviewed and blood taken for measurement of somatic mutations. Mutation rates for HPRT and glycophorin A were estimated with standard assay techniques. RESULTS: Homozygous mutations of glycophorin A were increased in underground miners (P = 0.0027) and the mutation rate tended to rise with increasing exposure with the exception of the highest exposure (> 10 working level months). However, there was no association between place of work and either the hemizygous mutations of glycophorin A or the HPRT mutation. CONCLUSIONS: There may be an association between glycophorin A mutations and previous occupational exposure to ionising radiation. However, not enough is known at present to use these assays as biomarkers for historical exposure in underground mining cohorts. PMID- 8704867 TI - Upper aerodigestive cancer in battery manufacturers and steel workers exposed to mineral acid mists. AB - OBJECTIVE: To assess the risk of cancer from inhalation of mineral acid mists. METHODS: A cohort study and nested case-control study of upper aerodigestive tumours were carried out in men employed since 1950 at two battery plants and two steel works in Britain. The cohort was identified from personnel records and included 2678 men with definite exposure to acid mists (mainly sulphuric acid), 367 with possible exposure, and 1356 who were unexposed. Mortality was compared with that in the national population by the person-years method. Cases of upper aerodigestive cancer were identified from death certificates and cancer registrations, and their exposure to acids was compared with that of age matched controls (five per case) from the same plant by conditional logistic regression. RESULTS: In follow up to 31 December 1993, 93% of men were traced, including 1277 who had died. Among the men definitely exposed to acid mists, overall mortality was less than in the national population (standardised mortality ratio (SMR) 0.92, 95% confidence interval (95% CI) 0.85-0.98) as was mortality from all cancers (SMR 0.92, 95% CI 0.79-1.05) and specifically from cancer of the larynx (SMR 0.48, 95% CI 0.01-2.70) and lung (SMR 0.98, 95% CI 0.78-1.22). A total of 15 incident or fatal cases of upper aerodigestive cancer were identified during follow up. When these men were compared with controls, risk was moderately increased in those who had worked for at least five years in jobs entailing exposures to sulphuric or hydrochloric acid in excess of 1 mg/m3 (OR 2.0, 95% CI 0.4-10). CONCLUSIONS: These findings are consistent with those from other studies which have indicated a hazard of upper aerodigestive cancer from acid mists. However, they indicate that any risk from exposures to sulphuric and hydrochloric acid below 1 mg/m3 is small. PMID- 8704868 TI - Work related respiratory symptoms in radiographers. AB - OBJECTIVE: To determine the prevalence of work related symptoms among radiographers compared with a control group of physiotherapists. METHOD: A postal questionnaire was used to collect information from radiographers and physiotherapists who registered in the United Kingdom during 1985-9. RESULTS: Satisfactory questionnaires were returned by 2354 (65%) of the radiographers and 3048 (69%) of the physiotherapists. There was a clear excess of work related symptoms among the radiographers. In particular, they were more likely to complain of symptoms that were worse at work, mouth soreness, sore, itchy, or runny eyes, persistent blocked nose, persistent itchy nose or sneezing, sore throat, headache, and of lower respiratory tract symptoms, which were also worse on workdays. These symptoms were associated particularly with the use of automatic processing machines. 235 radiographers gave a history of wheeze or chest tightness that had been worse at work or on days when at work. CONCLUSIONS: Work related symptoms suggesting irritation of the eyes and upper airways were more common in radiographers than controls, and may be related to exposure to x ray film processing chemicals. Men and women who reported work related wheeze or chest tightness will be followed up in more detail to assess the prevalence of occupational asthma in the cohort. PMID- 8704869 TI - Controlling the healthy worker survivor effect: an example of arsenic exposure and respiratory cancer. AB - OBJECTIVE: This investigation sought to examine whether methods proposed to control the healthy worker survivor effect would influence the shape or magnitude of the dose-response curve for respiratory cancer induced by arsenic. METHODS: Results from an unadjusted analysis are compared with results obtained by applying four different methods for control of the healthy worker survivor effect to data on arsenic exposure and respiratory cancer. The four methods are: exposure lag, adjustment for work status, cohort restriction, and the G null test. RESULTS: Cohort restriction gave erratic results depending upon the minimum years of follow up used. Exposure lag substantially increased the rate ratios and a non-linear shape (decreasing slope) compared with an unlagged analysis. Adjusting for work status (currently employed upsilon retired or otherwise not employed) yielded slightly higher rate ratios than an unadjusted analysis, with an overall shape similar to the baseline analysis. Results from the G null test procedure of Robins (1986), although not directly comparable with the baseline analysis, did show an adverse effect of exposure that seemed to reach a maximum when exposure was lagged between 10 and 20 years. CONCLUSIONS: All results confirm an adverse effect of arsenic exposure on respiratory cancer. In these data, it seems that the healthy worker survivor effect was not strong enough to mask the strong effect of arsenic exposure on respiratory cancer. Nevertheless, several methods show a stronger association between arsenic exposure and respiratory cancer after adjustment for the healthy worker survivor effect, suggesting that for weaker causal associations, studies not controlling for this source of bias will have low power to detect results. Although the G methods are theoretically the most unbiased, further work elucidating the validity of the assumptions underlying lagging, adjustment for work status, and the G methods are needed before clear recommendations can be made. PMID- 8704870 TI - Cancer incidence and specific occupational exposures in the Swedish leather tanning industry: a cohort based case-control study. AB - OBJECTIVE: To study the effect on the incidence of cancer of exposure to chemicals handled in the leather tanning industry. MATERIALS AND METHODS: A case control study was performed within a cohort of 2487 workers employed for at least six months during the period 1900-89 in three Swedish leather tanneries. 68 cancer cases (lung, stomach, bladder, kidney, nasal, and pancreatic cancers and soft tissue sarcomas) and 178 matched controls were studied. Effects of chemical exposures on cancer incidence, adjusted for age at risk, sex, and plant were estimated with a conditional logistic regression model. RESULTS: A significant association was found between exposure to leather dust and pancreatic cancer (odds ratio (OR) 7.19, 95% confidence interval (95% CI) 1.44 to 35-89). An association was indicated between leather dust from vegetable tanning and lung cancer. After adjustment for smoking habits a tentative association between organic solvents and lung cancer lost its significance. No association was found between exposure to chlorophenols and soft tissue sarcomas. CONCLUSIONS: The significant association between leather tanning and soft tissue sarcomas that was found in our previous cohort analysis could not be explained by exposure to chlorophenols. On the other hand a significant association was found between exposure to leather dust and pancreatic cancer, and exposure to leather dust from vegetable tanning was often present in cases with lung cancer. Due to the small numbers of cases, the results can, however, only lead to tentative conclusions. PMID- 8704871 TI - Creatine kinase activities in brain and blood:possible neurotoxic indicator of acrylamide intoxication. AB - OBJECTIVES: To examine whether the activities of creatine kinase (CK) correlate with neurological disturbances caused by acrylamide. METHODS: The activities of CK and other enzymes reported to be inhibited by acrylamide in the brain and plasma, and landing foot spread (LFS) were measured in mice and rats intoxicated with acrylamide. RESULTS: Activity of CK was suppressed by acrylamide in the brain of mice in parallel with the neurological dysfunction measured by LFS. No clear alterations were found in glyceraldehyde-3-phosphate dehydrogenase, neuron specific enolase, and lactate dehydrogenase activities over the experimental period (eight days for the exposure and 43 days for the recovery). In rats, among the plasma enzymes examined, suppression of CK activity was most notable, but thyroid activity was not affected. CONCLUSIONS: Among the enzymes so far examined, the CK activities in the brain and blood seem to be the most sensitive indicators of acrylamide intoxication. PMID- 8704872 TI - Relation of cumulative exposure to inorganic lead and neuropsychological test performance. AB - OBJECTIVES: To determine if measures reflecting chronic occupational lead exposure are associated with performance on neuropsychological tests. METHOD: 467 Canadian male lead smelter workers (mean (SD) age 43.4 (11.00) years, education 9.8 (3.18) years, years of employment 17.7 (7.43), and current blood lead concentration (B-Pb) 27.5 (8.4) micrograms dl-1) were given a neuropsychological screening battery. Time weighted average (TWA) and time integrated blood levels (IBL) were developed from B-Pb records obtained through regular medical monitoring (mean (range) TWA 40.1 (4.0-66.4) micrograms dl-1, mean IBL 765.2 (0.6 1625.7) micrograms-y dl-1). 14 neuropsychological variables were included in three multivariate analyses of covariance, with each exposure variable as the grouping variable (high, medium, and low) and age, education, score on a measure of depressive symptoms, and self reported alcohol use as the covariates. Groups did not differ in history of neurological conditions. RESULTS: Neither the B-Pb, TWA, nor IBL was significant by multivariate analyses of covariance (MANCOVA). When years of employment, a suppressor variable, was included as a covariate, IBL exposure groups differed significantly on digit symbol, logical memory, Purdue dominant hand, and trails A and B. CONCLUSIONS: A dose-effect relation was found between cumulative exposure (IBL) and neuropsychological performance at a time when current B-Pb concentrations were low and showed no association with performance. PMID- 8704873 TI - Neuropsychological verbal tests may lack "hold" properties in occupational studies of neurotoxic effects. AB - OBJECTIVES: In epidemiological studies of neurotoxic effects neuropsychological tests are often applied to assess possible functional changes. In these studies tests presumed to be resistant to neurotoxic effects, "hold tests", are often used to adjust performance in tests presumed to be sensitive to neurotoxic effects, "nonhold tests", in assessment of pre-exposure ability. This conception is based on experience from the examination of patients with organic brain disorders. For this adjustment to be valid hold tests should change comparatively little over time and should not change differently in groups of people with different exposures to neurotoxic agents. METHODS: These assumptions were examined in 71 carpenters and 135 painters divided in three subgroups according to level of cumulative exposure to organic solvents. The results were noted from two verbal tests (test A, which involved following verbal instructions and test C, word comprehension), which were performed at conscription (age 18-20). The tests resemble hold tests used in occupational studies. The same tests were performed again at the age of 45-60 together with a conventional synonym test (SRB 1), often used in occupational studies. RESULTS: In the three tests given at the time of the investigation some differences were found between the carpenters and the subgroups of painters. The painters with low exposure tended to perform better than the carpenters in all three tests and the heavily exposed painters tended to perform less well in the tests. These differences were not present at the age of 18-20. In one of the conscription tests painters with different cumulative exposure to organic solvents developed differently over time. There was little improvement among painters with heavy and intermediate exposure and obvious improvement among painters with low exposure. The results in the other conscription test showed similar tendencies. CONCLUSIONS: The results indicate that the hold tests examined do not meet basic criteria for hold tests used in occupational studies. Thus adjustments for conventional verbal tests seem inappropriate in such studies. It is possible that other presumed hold tests may meet the criteria but our results suggest that such tests also should be evaluated before being used as hold tests in working populations. PMID- 8704874 TI - Bacterial and fungal flora of dust deposits in a pig building. AB - OBJECTIVE: The purpose of the study was to investigate the bacterial and fungal flora of dust deposits in a newly built pig grower finisher building. Viable bacterial counts and microbial species found in a barn which had never housed pigs were compared with those in a barn housing 144 pigs. METHODS: The quantitative streak plate method was used to measure viable bacterial counts on nutrient agar or sheep blood agar. Viable bacterial counts of the dust deposits were expressed as the number of colony forming units (CFUs)/mg of dust. Gram positive cocci and Gram negative bacilli were identified by an automated system. Identifications with a confidence interval > 90% were accepted at the species level. Fungi were identified to the genus level with slide culture preparations on cereal agar. RESULTS: The lowest viable bacterial count (4.8 x 10(4)/mg of dust) was found in the barn with no pigs. In the barn with pigs the highest viable bacterial count (2.1 x 10(6)/mg of dust) was in dust from the top of a partition close to pig activity. Six species of bacteria or fungi were found in dust from the room with no pigs, whereas 22 different microorganisms were detected in dust from the room with pigs. With the exception of Enterobacter agglomerans no other species of the family Enterobacteriaceae was found in dust deposits in this new pig building. Twelve species of Gram positive bacteria were found in the room housing pigs. CONCLUSIONS: The pig is not only a source but also a disperser of airborne bacteria in pig buildings. Speciation of the microbial flora in dust from the pig building suggests that many of the microorganisms were either of human or environmental origin. Nevertheless as some of these microorganisms are known opportunistic pathogens or allergens and because of the documented increased incidence of chronic respiratory symptoms in pig workers, precautions to reduce inhalation of microbial or dust particles by pig workers seem prudent. PMID- 8704875 TI - Prognostic factors for return to work after a first compensated episode of back pain. AB - OBJECTIVES: To determine which factors measured at baseline and during the course of treatment influence time to return to work after a first compensated episode of back pain. METHODS: The design is a treatment inception cohort including 305 compensated workers out of 402 eligible ones presenting at two rehabilitation centres for conventional treatment. Crude and adjusted rate ratios (RRs) along with 95% confidence intervals (95% CIs) were estimated with the Cox's proportional hazards regression. RESULTS: 50% of workers had not returned to work after 112 days of follow up, and 11.3% still had not after 270 days. At the end of the study period (maximum follow up time was 1228 days), 230 workers (75.4%) had returned to work, 6.5% had not, and a similar percentage had retired, gone into vocational training, or returned to school. In the final model stratified for radiating pain during treatment, which was an important prognostic variable, workers between 21 to 30 years of age had a greater chance of returning to work (RR (95% CI) 1.43 (1.04 to 1.98) than those > or = 30. The other factors associated with a greater chance of returning to work were: a diagnosis of sprain or pain upsilon a diagnosis of intervertebral disc disorder (2.20 (1.23-3.91)), < 30 days of waiting between the accident and the beginning of treatment (1.30 (0.96 to 1.77)), a good flexion at baseline (1.52 (1.04 to 2.23)), absence of neurological symptoms during treatment (1.40 (0.98-2.00)), > 24 months of employment in the industry (1.49 (1.10 to 2.03)), working for a public industry upsilon a private one (1.63 (1.21 to 2.19)), and the ability to take unscheduled breaks (1.45 (1.06 to 1.97)). CONCLUSIONS: Even with a first episode of back pain, time to return to work is long and the proportion not returning is high. Return to work as expected is influenced by disease and host characteristics but also by social and work factors. Reinstatement programmes should account for all these factors. PMID- 8704876 TI - Exposure to cosmic radiation of British Airways flying crew on ultralonghaul routes. AB - British Airways has carried out radiation monitoring in Concorde for more than 20 years and has used a heuristic model based on data quoted by the National Aeronautics and Space Administration (NASA) to model radiation exposure in all longhaul fleets. From these data it has been calculated that no flight deck crew would exceed the control level of 6 mSv/y currently under consideration by regulatory authorities, which is three tenths of the occupational dose limit of 20 mSv/y recommended by the International Commission on Radiological Protection (ICRP). The model suggested that less than 4% of cabin crew based in Tokyo flying only between London and Japan could reach or exceed the 6 mSv/y level, based on a predicted effective dose rate of 7 microSv/h. To validate this calculation a sampling measurement programme was carried out on nine round trips flown by a Boeing 747-400 between London and Tokyo. The radiation field was measured with dosimeters used for routine personal monitoring (thermoluminescence dosimeters (TLDs) and polyallydiglycol carbonate neutron dosimeters). The limitations of the methodology are acknowledged, but the results indicate that the effective dose rate was 6 microSv/h which is consistent with the predicted effective dose rate of 7 microSv/h. This result, which is in accordance with other reported studies indicates that it is unlikely that any of the cabin crew based in Tokyo exceeded the 6 mSv/y level. In accordance with "as low as reasonably achievable" principles British Airways will continue to monitor flying crew routes and hours flown to ensure compliance. PMID- 8704877 TI - Exposure to styrene and mortality from nonmalignant respiratory diseases. AB - A cohort of 34,560 men and 6128 women employed in 660 European factories manufacturing reinforced plastic products, followed up originally to assess the risk of cancer, was used to assess the risk of non-malignant respiratory diseases associated with exposure to styrene. Mortality from pneumonia was associated with intensity of exposure to styrene, but this may have been due to chance. Mortality from bronchitis, emphysema, and asthma was not associated with styrene exposure. PMID- 8704878 TI - Surveillance systems and the role of a preventive medical team in chemical incidents. PMID- 8704879 TI - Factors affecting recognition of cancer risks of nuclear workers. PMID- 8704880 TI - Offspring sex ratio as an indicator of reproductive hazards. PMID- 8704881 TI - Why do patients with diabetic ketoacidosis have cerebral swelling, and why does treatment sometimes make it worse? PMID- 8704882 TI - Trends in age at the first medical evaluation of human immunodeficiency virus infection among infants born to infected mothers. New York City Pediatric Spectrum of HIV Disease Clinical Consortium. AB - OBJECTIVE: To evaluate the trends in age at the first medical evaluation of human immunodeficiency virus (HIV) infection among infants enrolled in the Pediatric Spectrum of Disease study born to mothers infected with HIV. DESIGN: Retrospective study based on medical chart review. SETTING: Nine pediatric centers in New York City. PARTICIPANTS: Infants (N = 925) born between January 1988 and December 1991 to mothers infected with HIV; the infants were examined for HIV infection by age 2 years and were receiving medical care. RESULTS: In each successive birth cohort, an increasing proportion of infants was examined by 3 months of age (from 35% in 1988 to 76% in 1991, chi 2 = 38.1, P < .001). The median age at the first evaluation persistently declined among the cohort evaluated by 24 months. The median age decreased from 6 months in 1988 to less than 1 month in 1991. The proportion of infants who were examined because of HIV related symptoms decreased in each successive birth cohort (1988, 65%; 1989, 59%; 1990, 42%; and 1991, 25%). CONCLUSIONS: An increasing proportion of newborns exposed to HIV are being examined within the first 3 months of life in 9 leading pediatric HIV centers in New York City. Prenatal HIV counseling and testing of mothers are optimal procedures because they benefit mothers, they allow the use of zidovudine to reduce the chance of HIV infection in the infants, and they allow mothers with HIV to be counseled about the potential risks of breast feeding. The family and the pediatrician must have knowledge of the infants' HIV status as early in life as possible to allow the necessary postnatal interventions, including Pneumocystis carinii pneumonia prophylaxis, which reduces morbidity and may prolong survival. PMID- 8704883 TI - Extremely low-birth-weight children and their peers. A comparison of school-age outcomes. AB - OBJECTIVE: To document 7-year developmental and educational outcomes in a cohort of predominantly white, middle-class, extremely low-birth-weight (ELBW, < 1000 g) children to address the incidence of increased developmental disability and the need for special educational services. DESIGN: Observational study. PATIENTS: Fifty-four ELBW children and 58 comparison children, who were matched for race, gender, and socio-economic status (30 with low birth weights [1500-2500 g] and 28 with birth weights > 2500 g). The ELBW cohort was drawn from 104 presurfactant survivors born between 1984 and 1986 and cared for in a single hospital. SETTING: Suburban, university-based tertiary referral center. MAIN OUTCOME MEASURES: Teachers' reports of classroom placement and special education services and tests of cognitive, motor, language, and visual-motor integration abilities were studied. RESULTS: Twenty-seven (50%) of 54 ELBW children were in regular classrooms with no special services compared with 21 (70%) of 30 in the low-birth weight group and 27 (93%) of 28 in the full-term group, indicating a significant trend toward increasing need for special services with decreasing birth weight across the 3 groups (P < .001). The ELBW group scored significantly lower than the comparison groups on all tests, although generally within the average range. Seventy-nine percent of ELBW children had average cognitive scores, but they averaged 14 to 17 points lower than the 2 comparison groups. Twenty percent of the ELBW children had significant disabilities including cerebral palsy, mental retardation, autism, and low intelligence with severe learning problems. CONCLUSIONS: Even with optimal socioeconomic environments, 20% of ELBW children are significantly disabled, and 1 of every 2 ELBW children requires special educational services. Objective testing pinpointed weakness on all measures compared with matched peer groups. PMID- 8704884 TI - Illicit substance use, gender, and the risk of violent behavior among adolescents. AB - OBJECTIVES: To analyze data from a nationally representative sample of high school students to investigate the relationship between substance use and violent behavior among adolescents and to examine this relationship in both male and female adolescents. DESIGN: Cross-sectional analyses of the 1991 Centers for Disease Control and Prevention's Youth Risk Behavior Survey. SETTING: Public and private schools in the 50 states. PARTICIPANTS: The participants were 12,272 high school students. MAIN OUTCOME MEASURE: To determine the prevalence of weapon carrying and physical fighting among male and female adolescents. RESULTS: A significant increase in the number of male and female adolescents carrying weapons and physically fighting was associated with all forms of substance use. Reports of carrying a weapon increased with recent alcohol consumption (34% vs 17%, P < .001) and use of marijuana (48% vs 22%, P < .001), cocaine (71% vs 25%, P < .001), and anabolic steroids (62% vs 25%, P < .001). The prevalence of physical fighting was also significantly higher among adolescents who used illicit substances than among adolescents who denied drug use. The risk of violent behavior increased significantly, and was of equal magnitude, for adolescent females and males who were illicit substance users. CONCLUSIONS: Alcohol and illicit substance use are highly associated with an increased risk of violent behavior. These data also demonstrate that the risk of violence by adolescent females who are substance users is substantial. PMID- 8704885 TI - Impact of maternal group B streptococcal screening on pediatric management in full-term newborns. AB - BACKGROUND: The American Academy of Pediatrics strategy to prevent early-onset neonatal sepsis with group B streptococcus (GBS) relies on maternal antepartum GBS cultures, while the American College of Obstetrics and Gynecology strategy does not. OBJECTIVE: To evaluate the impact of the 2 strategies on the care of asymptomatic full-term newborns. DESIGN/SETTING: Self-administered survey mailed to a national random sample of US pediatricians who were members of the American Academy of Pediatrics. PARTICIPANTS: A total of 461 members of the American Academy of Pediatrics who routinely care for newborns. MAIN OUTCOME MEASURE: Self report of diagnostic and treatment strategies for asymptomatic full-term newborns who were born under different clinical scenarios. Maternal risk factors, antepartum maternal GBS screening status, and maternal treatment with intrapartum antibiotics were varied across the scenarios. RESULTS: Pediatricians treating asymptomatic full-term newborns born to risk factor-negative mothers reported ordering tests (63.3% in GBS-positive cases vs 6.7% with GBS unknown; P = .001) and antibiotics (21.5% in GBS-positive cases vs 0.9% with GBS unknown; P = .001) more frequently when presented with a positive maternal GBS screening result. Maternal intrapartum treatment had little impact on pediatric practice when risk factors were absent. In risk factor-positive mothers, pediatricians reported an increase in their antibiotic usage in response to a positive maternal GBS screen (61.8% in GBS-positive cases vs 36.9% with GBS unknown; P = .001). In risk factor positive mothers with unknown results of GBS screening, use of intrapartum antibiotics increased the number of pediatricians who reported that they would prescribe antibiotic therapy. CONCLUSIONS: Obstetrical strategies to decrease the risk of neonatal GBS sepsis increase pediatric services provided to full-term healthy newborns. This increase in services by pediatric practices is likely to be greater with the screening-based strategy recommended by the American Academy of Pediatrics. PMID- 8704886 TI - Invasive pneumococcal infection in a community hospital, 1993 to 1995. Characteristics of resistant strains. AB - OBJECTIVES: To review all cases of invasive Streptococcus pneumoniae infection at a single institution and to identify factors that may allow distinguishing between penicillin-resistant pneumococcal (PRP) and penicillin-sensitive pneumococcal strains at presentation for emergency care. PATIENTS AND METHODS: Consecutive patient series of all children with positive blood and cerebrospinal fluid cultures for S pneumoniae from January 1993 to April 1995 at a general community hospital. RESULTS: Fifty-three patients with invasive S pneumoniae infections were identified; 21% of the infections were resistant to penicillin, 9% were resistant to multiple antibiotics, and 4% were highly resistant to penicillin (minimum inhibitory concentration, > 2 mg/L). At admission, a diagnosis of meningitis or sepsis was made in 14 patients; of these, 8 cerebrospinal fluid cultures yielded S pneumoniae, and 5 were PRP (P < .001). Significant differences between sensitive and resistant strains were not found for sex, age, race, ill household contacts, or physician type. Five of 11 children with PRP strains were either receiving antibiotics concurrently or in the prior 30 days compared with 5 of 42 children with penicillin-sensitive pneumococcal strains (P = .002). Children with PRP strains had a lower mean white blood cell count (14.9 x 10(9)/L vs 22.8 x 10(9)/L, P = .008) than children with penicillin-sensitive pneumococcal strains, owing to a lower mean absolute neutrophil count (7250 vs 12700, P = .006). Children with lower white blood cell and absolute neutrophil counts did not, however, differ in other objective measures. Regression analysis showed that the combination of current or prior antibiotic use and an absolute neutrophil count predicted 63% of the resistant strains and 92% of the sensitive strains (P = .001). CONCLUSIONS: A high proportion of S pneumoniae infections were caused by strains that were resistant to penicillin. We discuss factors that are associated with resistance in this population. The PRP strains were associated with recent antibiotic therapy and a low absolute neutrophil count. Further surveillance is warranted by these findings. PMID- 8704887 TI - Linking clinical and public health approaches to improve access to health care for socially disadvantaged mothers and children. A feasibility study. AB - OBJECTIVE: To test the feasibility of combining home- and office-based interventions to improve access to health care and health outcomes of Medicaid eligible mothers and infants. DESIGN: Randomized trial in 2 counties in North Carolina (1 rural, 1 urban). Information on health and developmental outcomes was obtained by face-to-face interviews, medical chart abstractions, hospital medical records, and state data tapes. PARTICIPANTS: Ninety-three Medicaid-eligible first time pregnant women in their third trimester and their subsequently born infants, who were followed up until they were 6 months old, and 3 pediatric practices and 1 family practice. INTERVENTIONS: Coordinated home visit and office intervention, office intervention, and usual care. Home visits by 3 public health nurses provided parental education and social support and linked families with needed community resources. Women in the office intervention group were encouraged to seek health care for their infants from one of the primary care practices. Participating offices received assistance with Medicaid billing, help developing a system to improve preventive care, and customized patient education materials. RESULTS: Mothers reported that the nurses helped them in areas related to the content of the program. An office system for prevention was developed and implemented in all 4 practices for study patients. Families in the intervention groups were more likely than control families to have had a prenatal visit with a pediatrician (P = .01, chi 2), a primary care office as the regular source of sick care (P = .02, chi 2), and less waiting time (P = .02, Student t test). They were also more likely to recall receiving patient education materials (P = .007, chi 2). CONCLUSIONS: It is feasible to link clinical and public health approaches to improve the quality and effectiveness of care for socially disadvantaged children. Such interventions should be tested in defined populations. PMID- 8704888 TI - The psychological and medical sequelae of war in Central American refugee mothers and children. AB - OBJECTIVE: To investigate the physical and mainly psychological sequelae of exposure to war in Central American children and their mothers who immigrated to the United States on average 4 years before the study began. DESIGN: Interview study. PARTICIPANTS: Twenty-two immigrant Central American women caretakers and 1 of their children aged 5 to 13 years. MAIN OUTCOME MEASURES: Standardized and new measures were administered to assess children's physical and mental health symptoms and exposure to political violence. RESULTS: Eighteen of the 22 children had chronic health problems. Fifteen children and all of the adults had observed traumatic events, including bombings and homicides. Thirteen of the children showed mental health symptom profiles above established norms, although only 2 met the criteria for posttraumatic stress disorder according to their own reports. Many of the caretakers were unaware of their child's psychological distress. Four of the mothers exhibited posttraumatic stress disorder, and their symptoms predicted their child's mental health. CONCLUSIONS: Pediatricians are sometimes the first and only contacts these families have with health care providers. Caretakers' reports of children's mental health are often incomplete. It is therefore important for physicians to probe for "hidden" symptoms in refugee children. These family members may need referrals to social and psychological services, and pediatricians can open the gates to existing community networks of support. Because we found that maternal mental health influences the child's, the child's interests are well served when pediatricians also encourage the mother to contact services for herself if she confides that she is experiencing some of the severe psychological sequelae reported by the women in this study. PMID- 8704889 TI - Pattern of prenatal care and infant immunization status in a comprehensive adolescent-oriented maternity program. AB - OBJECTIVE: To examine the relationship between patterns of prenatal care utilization and the subsequent pattern of preventive infant health care utilization among patients in a comprehensive, multidisciplinary, adolescent oriented maternity program. METHODS: We hypothesized that the mothers of incompletely immunized 8-month-olds were less compliant with their own prenatal care appointments than were mothers of fully immunized 8-month-olds. We retrospectively reviewed the medical records of 150 consecutively delivered infants and their adolescent mothers. Data concerning the pattern of prenatal and postnatal use of preventive health care services and potentially confounding maternal characteristics were collected. RESULTS: Of the 150 infants aged 8 months, 22 (14.7%) were incompletely immunized. Mothers of completely and incompletely immunized infants did not differ in age, school enrollment status, or compliance with prenatal appointments. However, the latter group initiated prenatal care later, obtained fewer prenatal visits, returned later for postpartum care, and were more likely to be black and to report inadequate family support after delivery. Three of the 5 characteristics entered a logistic regression function that predicted the risk of incomplete immunizations at 8 months of age: third-trimester initiation of prenatal care (odds ratio, 4.05; 95% confidence interval, 1.19-13.7), inadequate family support (odds ratio, 3.42; 95% confidence interval, 1.17-10.0), and black race (odds ratio, 3.14; 95% confidence interval, 1.19-8.69). The total model chi 2 was 15.8 (P < .001). CONCLUSIONS: Among patients in a comprehensive adolescent-oriented maternity program, the timing of the first prenatal visit helps to identify infants who are at increased risk for incomplete primary immunization status. Our findings favor preferential allocation of scarce, costly outreach services to infants born to adolescent mothers who enter prenatal care during the third trimester. PMID- 8704890 TI - Risk factors for the infant prone sleep position. AB - OBJECTIVE: To identify parental characteristics associated with infants being placed to sleep in the prone position. STUDY DESIGN: Cross-sectional survey. PATIENTS: Randomly selected King County, Washington, infants born on the same days as King County infants who died of sudden infant death syndrome between November 1992 and October 1994. METHODS: Parents of study infants responded to a telephone interview about sleep position in their infants. Parents were asked how they usually put their infants to bed during the previous 2 weeks, and if they were aware of any recent advice on sleep position in young infants. Demographic data were also collected during the telephone interview. Logistic regression was used to identify infant and parental characteristics associated with the prone sleep position. RESULTS: Parents of 178 infants were interviewed; 28.1% responded that their infants usually slept prone, 66.9% slept nonprone, and 5% had no usual sleep position. Parents who were unaware of sleep position advice were more likely to place their infants prone than those who were aware of this advice (odds ratio, 3.5; 95% confidence interval, 1.5-7.8). Among parents who were aware of sleep position advice, mothers younger than 20 years were more than 10 times as likely to place their infants prone than were older mothers (odds ratio, 10.7; 95% confidence interval, 1.1-107.0). For those who were unaware of sleep position advice, single mothers were more likely to place their infants prone (odds ratio, 14.0; 95% confidence interval, 1.5-133.2). Single mothers and parents of low birth-weight infants were more likely to be unaware of recent medical advice regarding optimal sleep position for infants. CONCLUSIONS: The results of this study may provide direction to future efforts to encourage nonprone sleeping. Knowledge of the risk is associated with decreased use of prone sleep position. Single mothers should be targeted for intensive educational efforts regarding the hazards of prone sleeping. Among teenage mothers, awareness of the association between prone sleeping and sudden infant death syndrome may not be adequate to change behavior; educational interventions that are more focused for this age group may be needed. PMID- 8704891 TI - Response of human immunodeficiency virus-exposed and -infected infants to Haemophilus influenzae type b conjugate vaccine. AB - OBJECTIVE: To evaluate the response of human immunodeficiency virus (HIV) infected and -exposed infants to the primary series and booster dose of Haemophilus influenzae type b (Hib) conjugate vaccine. DESIGN: Retrospective study. PATIENTS AND SETTING: The HIV-exposed and -infected infants who were attending the Special Immunology Family Clinic at The Children's Hospital of Philadelphia (Pa). MAIN OUTCOME MEASURES: Geometric mean antibody titers (GMTs) to Hib polyribosyl ribitol phosphate capsular antigen were assessed after the primary series and again after the 15-month booster doses. In addition, the percentages of patients who responded with polyribosyl ribitol phosphate antibody levels greater than both 0.15 and 1.0 mg/L were compared between groups. RESULTS: After the 3-dose primary series, the GMTs were lower in the HIV-infected infants compared with those in the HIV-exposed, uninfected infants (0.86 vs 2.30, P = .02). Forty-six percent of the HIV-infected infants mounted a response ( > 1.0 mg/L) compared with that in 79% of the HIV-exposed infants (P = .05). Among the HIV-infected infants, there was no difference in the GMTs based on CD4+ cell counts or HIV-related symptoms. After the 15-month booster dose, the GMTs were not significantly different in the HIV-infected and -exposed infants. As a group, the HIV-infected infants responded to the booster dose with a 2-fold increase in the GMTs, and significantly more of these infants had antibody concentrations above 1.0 mg/L compared with their response to the primary series (62% vs 38%, P = .02). CONCLUSIONS: Most of the HIV-infected infants responded to the primary series of Hib conjugate vaccine with antibody concentrations greater than 0.15 mg/L, but the GMTs were significantly lower than those in the uninfected infants. The primary series of Hib conjugate vaccine appeared to be capable of inducing specific immunologic memory in the HIV-infected infants. The HIV-infected infants had a significant response to a booster dose of Hib conjugate vaccine, as measured by using the GMTs and the percentage of infants with antibody concentrations greater than 1.0 mg/L. The duration of protective titers will need to be followed in this population of patients who are at a high risk for serious bacterial disease. PMID- 8704893 TI - Genital findings in adolescent girls referred for suspected sexual abuse. AB - BACKGROUND: Sexual abuse is a common problem affecting adolescent girls, but the frequency of medical findings in this population has not been specifically described. OBJECTIVE: To describe the frequency of specific genital findings in a group of pubertal girls who had experienced probable or definite sexual abuse. DESIGN: Patient series, medical chart and photograph review. SETTING: Specialty referral clinic for abused children. PATIENTS AND SELECTION: Referred sample of female patients, examined between January 1, 1987, and June 30, 1994, with Tanner genital stages 3, 4, or 5, who reported a history of penile-vaginal penetration, had colposcopic photographs taken, and were determined, by means of a previously described classification system, to have experienced probable or definite abuse. INTERVENTIONS: None. MAIN RESULTS: The study included 204 girls, aged 9 to 17 years (mean, 13 years); race or ethnicity was Mexican American in 57%, white in 34%, and other in 9%. Abnormal genital findings were documented in 32% of patients overall but were more common when the girls had reported bleeding at the time of the assault (50% vs 26%; P = .004, chi 2 analysis), or when the examination occurred within 72 hours of the last episode of abuse (69% vs 26%; P < .001, chi 2 analysis). Transections of the hymen were unusual (8%), but notches in the hymen were more common (25%). CONCLUSIONS: Normal or nonspecific results of genital examinations are commonly found in adolescents who have been sexually abused, unless the abuse was very recent. Further studies are needed to document the healing of genital injuries in victims of acute assault and the frequency of hymenal findings in nonabused, non-sexually active adolescents. PMID- 8704892 TI - Growth of Chicago-area infants, 1985 through 1987. Not what the reference curves predict. Pediatric Practice Research Group. AB - OBJECTIVE: To determine if the National Center for Health Statistics (NCHS) growth curves (including 867 white infants, born between 1929 and 1975, in the Yellow Springs, Ohio, area) reflect contemporary infant growth in pediatric practices. DESIGN: Observational cohort study of healthy term infants. Office personnel obtained standardized measurements at health maintenance visits. SETTING: Ten pediatric community practices that were members of the Chicago, III, area Pediatric Practice Research Group (PPRG). METHODS: Measurements of 1574 PPRG infants, seen on at least 5 occasions between 2 and 54 weeks of age, generated sex-specific growth curves using a 3-parameter mathematical model fitted to the serial data for each infant. Values from the computed curves were compared with NCHS growth references at 1, 3, 6, 9, and 12 months. The birth weights of PPRG and NCHS cohorts were compared. RESULTS: Mean birth weight of PPRG infants was significantly greater than a similar NCHS measure. Compared with the NCHS reference curve, PPRG infants were heavier at 1, 3, and 6 months, longer on all comparisons, and had greater head circumferences, particularly in the early months of age. CONCLUSIONS: The NCHS growth curves do not accurately reflect infant growth in this cohort. Nationally representative data are needed to revise the NCHS growth curves. PMID- 8704894 TI - Missed opportunities for vaccination and the delivery of preventive care. AB - OBJECTIVES: To evaluate the relative impact of 2 types of missed opportunities (MOs) for vaccination, acknowledged and unacknowledged, on the immunization status of children at 2 years of age and to measure the delivery of immunizations and health care maintenance (HCM) after these types of MOs. DESIGN: Case-control study. SETTING: A large multispecialty clinic serving primarily a managed care population in Tucson, Ariz. PATIENTS: Charts of 1165 patients, ages 2 to 4 years, were reviewed for immunization status by 2 years of age. Of these patients, 652 had received all of their medical care at the clinic during their first 2 years of life. The 76 patients found to be underimmunized (UI) at 2 years of age and 76 controls, who were fully immunized (FI) by 2 years of age, frequency matched for age, were studied. MAIN OUTCOME MEASURES: The charts were reviewed for the following information: sex, age, immunizations received, HCM visits, sick visits, MOs, and pediatrician-patient interchange regarding immunization status during visits. RESULTS: Of the children who received all their care at the study site, 88% were FI. The mean number of MOs per patient occurring during the first 2 years of life was 5.8 for the UI children and 2.6 for the FI children. Only an unacknowledged MO, defined as a visit when the patient's immunization status was not reviewed, was associated with having a deficient immunization status at 2 years of age (4.6 vs 1.7, P < .001). Acknowledged MOs did not differ between UI and FI children (0.95 vs 0.76, P = .67). Immunizations were received at the subsequent visit (within a month) twice as often following a visit at which the child's deficient immunization status was acknowledged and a plan for follow-up made (P < .001). However, HCM was often not completed at the return visit. Following acknowledged MOs, patients did not return as directed 25% of the time for UI children and 5% of the time for FI children. Only 30% of FI patients completed the number of HCM visits recommended by the American Academy of Pediatrics. CONCLUSIONS: In the population studied, lack of review of the patients' immunization status was the primary cause of MOs to vaccinate. Although patients returned significantly more frequently when informed of their child's deficient immunization status, poor follow-up remained a significant problem in children who were eventually found to be lacking immunizations at 2 years of age. PMID- 8704895 TI - Immunization status as determined by patients' hand-held cards vs medical records. AB - OBJECTIVE: To determine whether patients' hand-held immunization cards provide accurate assessments of immunization status when compared with their corresponding medical records. SETTING: Urban hospital emergency department immunization program. DESIGN: Comparison of 2 criterion standards. PATIENTS: Children aged 4 months to 6 years who presented consecutively with their immunization cards and received routine care in the hospital's pediatric clinic. SELECTION: Of 673 eligible patients seen in the immunization program from November 1992 to October 1993, 140 were randomly selected for comparison of immunization card and medical record immunization dates; in addition, all 123 eligible patients seen between August and October 1994 were selected. Of the total of 263 children, medical records for 257 (98%) were available for review. The dates of diphtheria-tetanus-pertussis, polio, measles-mumps-rubella, and Haemophilus influenzae type b immunization from immunization cards and medical records were recorded, as were patient age, sex, and ethnicity. Immunization card medical record immunization date pairs were compared. Each immunization card and medical record was categorized as up to date, due for immunization, or delayed 2 months or more for any immunization at the time of the visit. RESULTS: In 218 (85%) of 257 cases, the immunization card and medical record immunization dates were identical (McNemar test, P = .63). The immunization card and medical record agreed that patients were due for immunization in 91 cases and agreed that patients were not due for immunization in 138 cases (kappa = 0.77; 95% confidence interval, 0.70-0.85). The immunization card and medical record agreed that patients were delayed for 1 or more immunizations in 51 cases and agreed that patients were not delayed in 187 cases (kappa = 0.79; 95% confidence interval, 0.71-0.88). CONCLUSION: The hand-held immunization card is a suitable alternative to the medical record when the need for immunization is assessed or when rates of immunization delay in populations are determined. PMID- 8704896 TI - Evaluation of the 'Best Start' breast-feeding education program. AB - OBJECTIVE: To compare the frequency of breast-feeding before and after the "Best Start" breast-feeding educational program was implemented. DESIGN: Preintervention-postintervention trial. SETTING: A women's health center and children's hospital serving a low-income population. PATIENTS: Ninety mother infant pairs of whom the infants were born between January 2 and February 28, 1993, compared with 90 mother-infant pairs of whom the infants were born between January 2 and February 28, 1994. INTERVENTION: Best Start, a breast-feeding education program, was presented to health professionals and clerical staff of the Women's Health Center. At the first prenatal visit, a woman is asked, "What do you know about breast-feeding?" instead of "Are you going to breast-feed or bottle-feed this baby?" The program elicits and acknowledges the mother's concerns and then educates her about the benefits of breast-feeding. This is repeated at each prenatal visit. OUTCOME MEASURES: Sociodemographic data and breast-feeding rates of preintervention groups were compared with those of postintervention groups. Both hospital and out-patient records were reviewed. RESULTS: In 1993, 13 (15%) of 86 mothers breast-fed at hospital discharge compared with 25 (31%) of 81 in 1994 (P < .03). At the 2-week clinic visit, 11 (13%) of 86 were still breast-feeding in 1993 compared with 17 (21%) of 81 in 1994 (P > .20). The impact on mothers aged 19 years or less was particularly marked, with a tripling of the breast-feeding rate at hospital discharge from 11% (2/18) in 1993 to 37% (10/27) in 1994. CONCLUSION: This simple, low-cost educational intervention with a change in the manner of presentation significantly improved the breast-feeding rates of this low-income population. PMID- 8704897 TI - Radiological case of the month. Retinoblastoma presenting with orbital cellulitis. PMID- 8704898 TI - Picture of the month. Polyarteritis nodosa with angioedema. PMID- 8704899 TI - Pathological case of the month. Congenital cardiac rhabdomyoma. PMID- 8704900 TI - Should breast-feeding by women with silicone implants be recommended? PMID- 8704901 TI - Vitamin A and respiratory syncytial virus infection. PMID- 8704902 TI - Medical informatics and pediatrics. PMID- 8704903 TI - Immunization status of poor children. PMID- 8704905 TI - [The role of immunological changes in diseases in regions contaminated by radionuclides after the accident at the Chernobyl Atomic Electric Power Station]. AB - Autoimmune deviations, both humoral and cellular, related to antigens of thyroid gland, microsomes and thyroglobulin, were observed in residents of controlled districts of Bryansk and Tula regions of Russia. The importance of these deviations at hyperplasia of thyroid gland was demonstrated. In formation of cataracts under chronic influence of low doses of ionizing radiation the humoral autoimmune mechanisms are active but not the cellular ones. The increased content of antibodies against the antigens of crystalline lens found in the residents of the controlled regions shows the possibility to develop initial manifestations of cataract under the effect of low radiation doses. PMID- 8704904 TI - [Late changes in the body function of the inhabitants in the area of the eastern Urals radioactive trace]. AB - Using a mathematical methodology based upon the theory of pattern recognition, the adequacy of this methodology being confirmed by the corroborative results obtained with neuro-net non-parametric methods of analysis, it proved possible to discriminate reliably between generalized organism's statuses, as described with a big number of laboratory indices, of the inhabitants of different areas of a big industrial city which differ in levels of radioactive and non-radioactive environmental pollution. PMID- 8704906 TI - [The functional status of the immunocompetent cells in animals at different times after the chronic action of irradiation and heat]. AB - The proliferative response of thymic and splenic cells and the production of interleukin-1 and interleukin-2 by splenocytes were studied a day, three and six months after 25-fold separate and combination radiation (total dose 0.5 Gy) and heat (37 degrees C, 4h). The proliferative activity of lymphocytes of both organs was found to enhance a day after heating. No changes were found three months later. The proliferative activity of thymocytes in rats exposed to radiation and combination of the two factors was inhibited after six months. The production of interleukin-1 was lowered after irradiation. PMID- 8704907 TI - [The modification of a radiation lesion with 9a-homo-13-thiaprostanoids]. AB - The radioprotective properties of 12 compounds of 9a-homo-13-thiaprostanoid series were investigated under gamma irradiation using the molecular model of beta-carotene radio-oxidation in oleic acid in vitro, erythrocyte radiomimetic model in ex vivo-in vitro system as well as in vivo radiation damage in mice. Most of these compounds stimulated the radio-oxidation of beta-carotene, however in this model two prostanoids with natural alpha-chain displayed radioprotective properties. Expressed membrane stabilizing effect of two 9a-homo-13 thiaprostanoid nor-analogues was revealed in radiomimetic model experiments. Two 10, 10-dimethyl-13-thiaprostanoids raised animal radioresistance during in vivo experiments. PMID- 8704908 TI - [The effect of low doses of a synthetic analog of a quinoid radiotoxin on the viability of normal and gamma-irradiated animals]. AB - The method for a production of synthetic quinoid radiotoxins in vitro has been developed and described. Synthetic quinoid radiotoxins like quinoid radiotoxins (qRT) which are being produced from irradiated tissues of the organisms have demonstrated high toxicity at relatively high qRT concentrations. However, when synthetic qRT is introduced into the organisms in ultra-small concentrations, one can observe the opposite action: the resistance of the organism increases and a number of essential functions are activated. Quinoid radiotoxins are assumed to take part in regulatory processes responsible for radiation hormesis. PMID- 8704909 TI - [The effect of sodium nucleinate on the precursors of antigen-dependent suppressors following irradiation]. AB - Sodium nucleinate was administered perorally to mice in the dose of 30 mg/kg. After the treatment the suppression activity of spleen cells decreased. The decrease was more pronounced in mice exposed to the doses of 0.5 and 1 Gy than in intact mice. The dose of 240 mg/kg did not affect suppression activity. PMID- 8704910 TI - [The combined action of ionizing radiation and stress on antibody formation in mice]. AB - Disturbances in humoral immune response to sheep erythrocytes after separate and combined effect of ionizing radiation (2 and 4 Gy) and stress (swimming for 10 of 60 min) was studied in mice. The increase in sensitivity to stress was found in irradiated mice. Superposition of undulating dynamics of post-stress immunosuppression on dynamics of post-radiation disorder was revealed. This is due to the different mechanisms of disturbances: redistribution of precursors of immunocompetent cells between immune organs in the first case and destruction of cells in the second case. PMID- 8704911 TI - Therapeutical effect of silymarin on nucleic acids in the various organs of rats after radiation injury. AB - The therapeutical effect of the hepatoprotective drug silymarin (Flavobion) was investigated in rats after total body gamma irradiation with a dose of 6 Gy. Silymarin (70 mg kg-1 p.o. by tube) was administered twice a day over 7 and 14 days after irradiation. At the end of therapy, the effect of silymarin was evaluated on the basis of quantitative changes of nucleic acids in the liver (regenerating after partial hepatectomy), spleen and bone marrow. It was found that the nucleic acid changes in irradiated rats were alleviated by the post radiation application of silymarin in its target organ--the liver, but also in the spleen and bone marrow. We assume that the therapeutical effect of silymarin on radiation induced changes of nucleic acids in various tissues of rats is caused mainly by the activation of cellular metabolism including the metabolism of nucleic acids. PMID- 8704912 TI - [An experimental study of the early changes in the body after simultaneous radiation exposure at a low dose and stress]. AB - In experiments in rats it was shown that simultaneous effect of low dose radiation (12.9 cGy) and stress (immobilization) resulted in early adaptation like changes (decrease in body weight, increase of adrenal gland weight, decrease in thymus weight, slowing down of blood coagulation). At the same time irradiation of immobilized animals prevented stressogenic disturbances such as depression of antiaggregation activity of vascular wall and decrease in leucocytes level. This was accompanied by the acceleration of mobilization of compensatory properties, that presumably is a basis for positive effects under combined influence. PMID- 8704913 TI - [The effect of gamma irradiation under different regimens on isolated and contacting cells of a HeLa culture]. AB - It was shown that isolated and united HeLa cells in culture responded to single irradiation and repeated low dose irradiation by different ways. The effect on radiosensitivity was revealed in united cells only. PMID- 8704914 TI - [Radiation-induced changes of alginic acid and sodium alginate]. PMID- 8704915 TI - [The hormonal status and carbohydrate-energy metabolism of rats after the long term action of low doses of ionizing radiation and heat]. AB - Hormonal status (blood content of triiodothyronine, thyroxin, insulin, 11 hydroxycorticosteroids), dehydration in the Krebs cycle, and activity of the first enzyme in the pentose-phosphate cycle, glucose-6-phosphate dehydrogenase, in the brain and myocardium of white rats were studied at different time periods after separate and combined prolonged exposure to radiation in relatively small doses and heat. It was found that combination of ionizing radiation and heat led to hypofunction of the endocrine glands and inhibition of dehydration processes in the Krebs Cycle. PMID- 8704916 TI - [Chemiluminescence, blood lipid peroxidation and neutrophil activity during the hypoxic training of persons subjected to ionizing radiation exposure]. AB - Free radical processes and some indices of antibacterial defense system have been examined in 29 male residents of Chernobyl area during adaptation to periods of intermittent hypoxia. 18 men (the experimental group) were exposed to normobaric isocapnic progressive hypoxia during 10 days of three daily 5-7 min sessions with 15 min breaks, and 11 men (control group) were exposed to air breathing. All subjects were divided into two subgroups with initial high (1) and low (2) level of blood chemiluminescence (ChL). Patients of the 1 subgroup were characterized with high oxygen-generated activity of neutrophils (OGA) and high malondialdehyde (MDA) concentration. After hypoxic training (HT) there was a decrease of spontaneous and initiated ChL and MDA. Patients of the 2 subgroup were characterised with low level both spontaneous and initiated ChL, low MDA concentration and low phagocytosing activity of neutrophils. After HT there was significant rise of initiated ChL and MDA concentration up to normal level with the increasing of neutrophil phagocytosing activity. We suggest that HT causes the normalizing effect on free radical processes in subjects who were exposed to radiation influences. PMID- 8704918 TI - [The effects of the simultaneous combined irradiation of diploid yeast cells of different genotypes using radiation with different LETs]. AB - Effects of simultaneous irradiation of diploid yeast cells with various genotypes with alpha-particles, gamma-rays or fast electrons were studied. When contribution of sparsely ionizing radiation to the overall dose is small (up to 25%) antagonism was observed, while at the 95% contribution there seems to be a tendency towards synergistic effect; at 50% contribution the results of the simultaneous combined irradiation do not contradict hypothesis on independent effect of radiation with different LET, which also implies a change in the RBE of simultaneous irradiation. PMID- 8704917 TI - [The relationship between respiratory system reactivity and neutrophil metabolism in hypoxia in persons subjected to ionizing radiation exposure]. AB - The group of 18 men exposed to radiation during amelioration work in the Chernobyl NPP was examined in the course of adaptation to intermittent hypoxia (rebreathing technique during 10 days of three daily 5-7 min sessions with 15 min break). The starting level of ventilatory response to hypoxic stimulus (HVR) did not differ from the one in persons living in non-contaminated areas. This hypoxic training (HT) caused the increase of HVR, activity of NADPH-oxidase and cationic protein content in neutrophils as well as various changes in myeloperoxidase activity. The correlation between respiration reactivity and deviations in neutrophil metabolism under HT was found. PMID- 8704919 TI - [The kinetics of radionuclide distribution between the plasma and formed elements of the blood in dogs]. AB - The investigation of microkinetics of 90Sr, 137Cs and 32P penetration into erythrocytes of dog's blood was made. It was shown that processes of penetration of 90Sr and 32P are monomolecular. The slow stage is monomolecular destruction of complexes of 90Sr and 32P with plasma components. The process of 137Cs penetration is bimolecular. The slowest stage of 137Cs penetration is adsorption on the surface of erythrocytes. PMID- 8704920 TI - [Radionuclide biokinetics as a function of the age of experimental animals]. AB - Behaviour of radionuclides with different distribution types in the organism of animals of various ages was studied. Depending on the age of experimental animals, an essential difference in biokinetic parameters such as coefficients of absorbtion from gastro-intestinal tract, distribution and retention levels in critical organs, retention and excretion rates of radionuclides was found. PMID- 8704922 TI - [The effect of the intratracheal administration of americium-241 on rat alveolar macrophages]. AB - In experiments in rats it was found that 241Am transitory decreases the total cell number and alveolar macrophage's percentage in bronchoalveolar lavage fluid (BALF): increases the macrophages size and nuclear size; and increases acid phosphatase and lactate dehydrogenase activities in BALF. It was suggested that 241Am causes and activation in the alveolar macrophages which probably appears as one of factors provoking lung injuries. PMID- 8704921 TI - [The effect of algisorb on the level of the accumulation of zirconium, ruthenium, iodine and cesium radioactive isotopes in the body of rats]. AB - The sorption effect of Algisorbum has been studied in rats following single and multiple intragastric administration. Algisorbum doses of 250-2000 mg/kg decrease the absorption of 106Ru and 95Zr by 50%, that of 137Cs by 15% and have no effect on 131I absorption. Application of a complex of agents to protect the body from nuclear fission products is discussed. PMID- 8704923 TI - Elimination of n-butylated hydroxytoluene methylation during fatty acid analysis by gas chromatography. AB - An improved method for fatty acids analysis with optimum recovery of highly polyunsaturated fatty acids methyl esters in biological systems is presented. The method is based on transesterification of phospholipid and triacylglycerols to fatty acid methyl esters using a commercially available reagent, Methyl-Prep II. Without proper precautions, as much as 50% of n-butylated hydroxytoluene (BHT) added to prevent oxidation of polyunsaturated fatty acids, could be methylated during the transesterification step. Methylated BHT elutes close to 14:0 (myristic acid) and no longer functions as an antioxidant, but the modified conditions virtually eliminate the methylation of BHT. Sample extraction and methylation was completed in 30 min at room temperature. A chelator (diethylenetriamine-pentaacetic acid; DTPA) is also added to prevent peroxidation of metal catalyzed free radical chain reactions. The standard deviations of the major fatty acids from multiple human plasma samples prepared on different days were less than 5%. The recovery of arachidonic acid, 20:4, from plasma was improved using the new method, and the recovery for docosahexaenoic acid, 22:6, spiked to human plasma was found to be 99%. PMID- 8704924 TI - Thin-layer chromatography and high-performance liquid chromatography for the assay of fatty acid compositions of individual phospholipids in platelets from non-insulin-dependent diabetes mellitus patients: effect of eicosapentaenoic acid ethyl ester administration. AB - Eight major phospholipids were separated by a TLC method with a one-dimensional developing system without any pretreatment of the plate and the fatty acids incorporated into each phospholipid class were analysed by an improved HPLC method with a simple elution system, which has advantages with respect to resolution and analysis time. The fatty acid compositions of individual phospholipids in platelets were investigated following administration of ethyl cis-5,8,11,14,17-eicosapentaenoate for more than 13 weeks to patients with non insulin-dependent diabetes mellitus. The cis-5,8,11,14,17-eicosapentaenoic acid compositions of all phospholipid classes were significantly increased with decreasing platelet aggregation rates after the administration. These results suggested that the present method provides the complete separation of individual phospholipids in sufficient amounts to allow fatty acid analysis on the isolated phospholipid moieties. PMID- 8704925 TI - Determination of retinoids by reversed-phase capillary liquid chromatography with amperometric electrochemical detection. AB - A method for separating and detecting retinoids by reversed-phase capillary liquid chromatography with amperometric electrochemical detection is described. Packed columns with an inner diameter of 180 microns were employed for the separation using C18 stationary phase and a mobile phase containing acetonitrile water methanol (65:32.5:2.5, v/v/v) with 1% tetrabutylammonium perchlorate and 0.174 M acetate buffered at pH 5. The detection cell consisted of a carbon fiber barrel electrode held at 0.9 V versus an Ag/AgCl reference. Injection volumes of 2 microliters produced detection limits of 2.73, 0.472, 0.428, and 0.267 fmol (or 410, 64.1, 60.9, and 38.2 pg ml-1) for 13-cis-retinoic acid, all-trans-retinoic acid, retinaldehyde, and retinol, respectively. This represents an improvement in detection limits of at least three orders of magnitude for similar analyses using liquid chromatography and UV absorbance detection. The detector signal was linear over two orders of magnitude of analyte concentration. Retinoid concentrations in bovine serum were determined and found to be in good agreement with previously reported values. PMID- 8704926 TI - Determination of lactate dehydrogenase isoenzymes in single lymphocytes from normal and leukemia cell lines. AB - This work demonstrates that our previously developed technique for single erythrocyte analysis by capillary electrophoresis with laser-induced fluorescence detection (CE-LIF) can be applied to study individual lymphocytes, with some modification in the cell lysing procedure. A tesla coil was shown to be capable of lysing the lymphocyte cells inside the capillary. The electromagnetic field induced by the tesla coil was believed to be responsible for breaking the cell membrane. The lactate dehydrogenase (LDH) isoenzyme activities and relative ratios between different LDH isoenzymes were measured for normal lymphocytes as well as B-type and T-type acute lymphoblastic leukemia cells. Both the LDH activity and the isoenzyme ratios show large variations among individual cells. The former is expected due to variations in cell size. The latter implies that single-cell measurements are less useful than the average values over a cell population as markers for leukemia. PMID- 8704927 TI - Hair analysis for nordiazepam and oxazepam by gas chromatography--negative-ion chemical ionization mass spectrometry. AB - A procedure is presented for the identification of nordiazepam and its metabolite, oxazepam, in human hair. The method involves decontamination of hair with dichloromethane, incubation in phosphate buffer (pH 7.6) in the presence of deuterated internal standards, liquid-liquid extraction, derivatization with N,O bis(trimethylsilyl)trifluoroacetamide plus 1% trimethylchlorosilane and gas chromatography-mass spectrometry using negative-ion chemical ionization with methane. Among thirty samples obtained from polydrug abusers, thirteen tested positive for nordiazepam, in the range of 0.25-18.87 ng/mg. Five samples were also positive for oxazepam, in the range 0.11-0.50 ng/mg. PMID- 8704928 TI - Determination of delta 9-tetrahydrocannabinol from human saliva by tandem immunoaffinity chromatography--high-performance liquid chromatography. AB - Smoking or ingestion of cannabis causes cognitive, perceptual and behavioural changes, which are responsive for impaired performance in driving motor vehicles. In this paper a novel liquid chromatographic assay for the selective quantification of delta 9-tetrahydrocannabinol, the major indicator of a present cannabis intoxication in saliva, is described. The method involves a column switching procedure and requires an extremely simple pre-treatment of the sample. Deproteinized saliva was directly injected into the chromatographic system. The clean-up and enrichment procedure was performed in an immunoaffinity column, followed by the transfer of the antigens to an octysilica analytical column. The immunoaffinity sorbent was obtained by covalent immobilization of specific antibodies on epoxy-activated silica. The mobile phase consisted of methanol aqueous 0.15 mol/1 NaCl solution (elution programmed) and the analyte was detected by measuring the UV absorption at 220 nm. Using an injection volume of 4.5 ml (dilution 3:2, v/v) the limit of quantification was 20 ng/ml, at a signal to-noise ratio of 5. Recoveries were estimated to be in the range of 70%. Both intra- and inter- day coefficients of variation were below 5%. PMID- 8704929 TI - Fast liquid chromatographic determination of urinary trans,trans-muconic acid. AB - trans, trans-Muconic acid (1,3-butadiene-1, 4-dicarboxylic acid, MA), a minor urinary metabolite of benzene exposure, was determined, after clean-up by solid phase anion-exchange chromatography, by reversed-phase HPLC on a C18 column (5 x 0.46 cm I.D., 3 microns particle size), using formic acid-tetrahydrofuran-water (14:17:969) as mobile phase and UV detection at 263 nm. The recovery of MA from spiked urine was > 95% in the 50-500 microgram/l range; the quantification limit was 6 micrograms/l; day-to-day precision, at 300 micrograms/l, was C.V. = 9.2%; the run time was less than 10 min. Urinary MA excretion was measured in two spot urine samples of 131 benzene environmentally exposed subjects: midday values obtained in non-smokers (mean +/- S.D. = 77 +/- 54 micrograms/l, n = 82) were statistically different from those of smokers (169 +/- 85 micrograms/l, n = 30) (P < 0.0001); each group showed a statistically significant increase between MA excretion in midday over morning samples. Moreover, in subjects grouped according to tobacco-smoke exposure level, median values of MA were positively associated with and increased with daily smoking habits. PMID- 8704930 TI - High-resolution anion-exchange and partition thin-layer chromatography for complex mixtures of 32P-postlabeled DNA adducts. AB - 32P-Postlabeling has emerged as a major tool for detecting DNA adducts resulting from exposure to complex carcinogen mixtures. An integral component of this assay is multi-directional PEI-cellulose TLC in which lipophilic 32P-adducts are resolved in high-salt, high-urea solvents following removal of the bulk of non adduct radioactivity. This TLC system is very effective for adducts formed following exposure to individual carcinogens; however, adducts resulting from exposure to complex mixtures (e.g. cigarette smoke) generally appear in the form of the so-called diagonal radioactive zones. By using mixtures of polycyclic aromatic hydrocarbon- and aromatic amine-DNA adducts as well as adducts in mouse skin treated with cigarette smoke condensate, we have demonstrated that a combination of 0.3-0.4 M NH4OH and isopropanol-4 M NH4OH (1-1.4:1) solvents can provide more sharply defined adduct spots than the commonly used urea solvents. The non-urea solvents also result in excellent resolution of many adducts which otherwise may remain buried in diagonal radioactive zones when using the urea solvents. In addition, the signal-to-noise ratio is increased 2- to 5-fold over the urea solvents enabling detection of discrete adducts at < or = 3 adducts per 10(10) nucleotides. These partition TLC solvents also involve fewer manipulations (e.g. no water washes to remove salt and urea), and are likely to be more informative with regards to the type of individual adducts detected in the biomonitoring of humans than has hitherto been possible. PMID- 8704931 TI - Capillary high-performance liquid chromatography-fast atom bombardment mass spectrometry of 24 cephem antibiotics. AB - Using capillary high-performance liquid chromatography (HPLC)-fast atom bombardment (FAB) mass spectrometry (MS), both positive and negative FAB mass spectra of 24 cephem antibiotics with diethanolamine (DEA) and glycerol (GLY) as matrices are presented. In the positive mode, an internal quasi-molecular peak together with relatively abundant fragment peaks were obtained from all 24 drugs with both matrices, though DEA provided more information on molecular mass of a compound than did GLY for some drugs. In the negative mode, the background was generally lower than that in the positive, but neither the quasi-molecular nor molecular peak was detected in several drugs with either matrix. The drugs were isolated from serum samples using an octadecyl reversed-phase cartridge; recoveries were generally over 60%, With this isolation and the capillary HPLC FAB-MS in the positive mode, ceftriaxone and cefazolin, two of the most popular cephem antibiotics, were successfully identified in 0.5 ml of sera obtained from a clinical or an autopsy case. PMID- 8704932 TI - Application of reversed-phase liquid chromatography and prepacked C18 cartridges for the analysis of oxytetracycline and related compounds. AB - The reversed-phase (RP) chromatographic separation of oxytetracycline (OTC) 4 epioxytetracycline (4-epiOTC), alpha-apooxytetracycline (alpha-apoOTC), and beta apooxytetracycline (beta-apoOTC) has been accomplished on an Inertsil C8 column at ambient temperature. Using the simplex method of solvent optimization, a 0.1 M ammonium acetate buffer (pH 3.0)-acetonitrile-tetrahydrofuran (72.5:12.5:15, v/v/v) mobile phase was found to give excellent separation of the compounds. OTC, 4-epiOTC, alpha-apoOTC and beta-apoOTC were resolved in 35 min with calculated detection limits of 40, 20, 50 and 140 ng/ml, respectively. Solid-phase extraction (using RP C18 cartridges) of OTC and OTC degradation compounds from distilled water and porcine muscle was tested at four concentration levels ranging from 200 to 2000 ng/ml (g); overall mean recovery of OTC from distilled water and porcine tissue was greater than 90% and 70%, respectively. PMID- 8704933 TI - Liquid chromatographic assay for a butenolide endothelin antagonist (PD 156707) in plasma. AB - A sensitive and selective liquid chromatographic assay for determining the non peptide endothelin A receptor antagonist PD 156707 (I) in rat plasma has been developed and validated. The analyte was isolated from matrix by solid-phase extraction. Liquid chromatographic separation was achieved isocratically on a 3.2 mm I.D., ODS column with a mobile phase of acetonitrile-ammonium phosphate (50 mM, pH 3.5) (44:56, v/v). Column effluent was monitored fluorometrically. Peak height ratios (analyte/IS) were proportional to I concentrations in rat plasma from 25 to 1000 ng/ml. Assay precision and accuracy for I, based on quality controls, was 9.5% relative standard deviation, with relative error of +/- 6.5%. The quantitation limit was 25 ng/ml for a 200-microliters sample aliquot. PMID- 8704934 TI - Simultaneous high-performance liquid chromatographic determination of caffeine and theophylline for routine drug monitoring in human plasma. AB - An HPLC method for the simultaneous determination of both caffeine and theophylline in human plasma is described, using a reversed-phase chromatography column, heated by a thermostatic oven at 35 degrees C, with UV detection and isocratic elution. The linearity and reproducibility of the method are verified. For the two drugs, the limit of detection is 0.1 microgram ml-1. This analytical method is rapid and reliable and allows routine controls of therapeutic levels of theophylline and caffeine, especially in premature infants where the volume of plasma samples is very small. PMID- 8704935 TI - Sensitive high-performance liquid chromatographic assay for pilocarpine in biological fluids using fluorescence derivatisation. AB - A sensitive assay for pilocarpine in biological fluids has been developed involving HPLC of a fluorescent derivative of 4-bromomethyl-7-methoxycoumarin. Pilosine as internal standard was added before the derivatisation step. The fluorescent derivatives were well resolved and separated from excess reagent and endogenous compounds on a cyanopropyl silica column. The detection limit of pilocarpine in biological fluids was 1.0 ng/ml and the assay was linear up to a concentration of 150 ng/ml. The assay was applied to a preliminary study of pilocarpine disposition in man after a single oral dose. This is the first report of pilocarpine excretion into saliva. PMID- 8704936 TI - Determination of diphenylpyraline in plasma and urine by high-performance liquid chromatography. AB - A rapid, reliable and specific reversed-phase high-performance liquid chromatographic procedure is described for the determination of diphenylpyraline hydrochloride at nanogram concentrations in plasma and urine. After extraction of the drug with n-pentane-2-propanol (50:1) from alkalinized samples, the organic extract was evaporated to dryness, reconstituted with methanol and chromatographed using a 5-micron Asahipak ODP-50 C18 column with UV detection at 254 nm. The elution time for diphenylpyraline was 7.9 min. The overall recovery of diphenylpyraline from spiked plasma and urine samples at concentrations ranging from 53 to 740 ng/ml were 94.65% and 92.29%, respectively. Linearity and precision data for plasma and urine standards after extraction were acceptable. The limit of detection was 15 ng/ml for both plasma and urine samples at 0.002 AUFS. PMID- 8704937 TI - Chiral separation of 10,11-dihydro-10, 11-trans-dihydroxycarbamazepine, a metabolite of carbamazepine with two asymmetric carbons, in human serum. AB - Chiral separation of 10, 11-dihydro-10, 11-trans-dihydroxycarbamazepine (CBZ diol), a metabolite of carbamazepine (CBZ) with two asymmetric carbons, in serum taken from epileptic patients receiving CBZ alone for a long period, was performed by high-performance liquid chromatography using a polysaccharide stationary phase with n-hexane-ethanol (75:25, v/v) as the mobile phase. The enantiomeric ratio (S,S-/R,R-CBZ-diol) was 10.74 +/- 1.13 (mean +/- S.D.), which could demonstrate the presence of the stereospecificity in the hydrolysis of 10, 11-dihydro-10, 11-epoxycarbamazepine (CBZ-epoxide) to CBZ-diol and/or in the conversion of CBZ-diol to some metabolite such as 9-hydroxymethyl-10 carbamoylacridan. This is the first paper to report the determination of each enantiomer and the enantiomeric ratio of CBZ-diol in serum of epileptic patients who received CBZ. PMID- 8704938 TI - Simple high-performance liquid chromatographic separation of oxazepam and its diastereoisomeric glucuronides in serum. Applications in a pharmacokinetic study in sheep. AB - This paper describes a highly specific and sensitive method for quantifying oxazepam and its diastereoisomeric glucuronides in serum. The method involves sample clean-up by solid-phase extraction on C18 cartridge followed by quantitation on a reversed-phase HPLC column. Diazepam is used as internal standard. Extraction recovery from serum proved to be more than 86%. Precision, expressed as C.V., was in the range 1.2-9.5%. The limits of quantification were 40, 400, and 200 nmol/l for oxazepam, S-(+)- and R-(-)-glucuronides, respectively. This method was applied to the determination of oxazepam and its diastereoisomeric glucuronides in serum collected during a pharmacokinetic study performed in sheep after oral administration of racemic oxazepam. S-(+)/R-(-) ratios were measured all along the sampling time collection and the pharmacokinetic parameters were determined. PMID- 8704939 TI - Determination of isonicotinic acid in the presence of isoniazid and acetylisoniazid. Studies on isonicotinic acid formation from isoniazid in isolated rat hepatocytes. AB - In comparison with the hepatocytes obtained from intact rats and rats pretreated with phenobarbital or 3-methylchoranthrene, the amount of isonicotinic acid (INA) formed from isoniazid (INH) increased substantially after incubation at 37 degrees C using the pretreated hepatocytes. This suggests an oxidative pathway for INA formation from INH, apart from hydrolysis. In order to explore the exact mechanism of INA formation in the hepatocytes, an HPLC assay for INA in the presence of INH and acetylisoniazid was developed. In this assay, INA was extracted after the preparation of an ion pair with tetra-n-butylammonium hydroxide, and analysed using an ODS column and a mobile phase consisting of 0.067 M potassium dihydrogenphosphate solution-methanol (96:4, v/v). The method is simple, accurate and especially suitable for INA determination after incubation of INH in isolated rat hepatocytes. PMID- 8704940 TI - Selective high-performance liquid chromatographic determination of artesunate and alpha- and beta-dihydroartemisinin in patients with falciparum malaria. AB - A novel solid-phase extraction and a robust high-performance liquid chromatographic (HPLC) separation procedure for artesunate and alpha- and beta dihydroartemisinin, using post-column alkali decomposition and UV detection is described. Extraction was performed with Bond-Elut Phenyl solid-phase extraction cartridges and analysis by HPLC was carried out using a Waters Symmetry C8 5 microns 150 x 3.9 mm I.D. column. The mobile phase was 50% acetonitrile in 0.1 M acetate buffer (pH 4.8) delivered at a flow-rate of 0.7 ml/min. The column eluate was mixed with 1.2 M potassium hydroxide in 90% methanol delivered at 0.3 ml/min, in a 1-ml reaction coil at 69 degrees C, to form UV-absorbing chromophores which were detected at 290 nm. The recovery of all analytes was greater than 80%. There was no significant difference in the peak-area ratio of alpha- and beta dihydroartemisinin in plasma. Preliminary pharmacokinetic data from six adult Vietnamese patients who received 120 mg of artesunate by intravenous injection for the treatment of acute falciparum malaria are presented. Despite limited data, the mean half-life of artesunate was approximately 3.5 min while that for dihydroartemisinin was 34 min. These data confirm the relatively rapid clearance of both artesunate and its principle active metabolite, dihydroartemisinin. PMID- 8704941 TI - Specific high-performance liquid chromatographic assay with ultraviolet detection for the determination of 1-(2-chloroethyl)-3-sarcosinamide-1-nitrosourea in plasma. AB - A facile, sensitive and highly specific HPLC method for assaying 1-(2 chloroethyl)-3-sarcosinamide-1-nitrosourea (SarCNU) in plasma has been developed. The drug was efficiently isolated from plasma by extraction with tert.-butyl methyl ether. A structurally related compound with similar physicochemical properties served as the internal standard (I.S.). Following evaporation of the organic solvent, the extract was reconstituted with 0.05 M ammonium acetate buffer, pH 5.0, and loaded onto a 4 micron Nova-Pak C18 column (15 cm x 3.9 mm), which was preceded by a 7 micron Brownlee RP-18 precolumn (1.5 cm x 3.2 mm). Chromatography was performed at ambient temperature using a mobile phase of methanol-0.1 M ammonium formate buffer, pH 3.7 (25:75, v/v). UV absorbance of the effluent was monitored at 240 nm. A flow-rate of 1.0 ml/ min was used for analyzing mouse and dog plasma extracts. Under these conditions, the drug eluted at 4.0 min and was followed by the I.S. at 6.1 min. An automatic switching valve was employed to allow the precolumn to be flushed 1.5 min into the run, without interrupting the flow of the mobile phase to the analytical column, thereby preventing the apparent build-up of extractable, strongly retained, UV-absorbing components present in mouse and dog plasma. Operating in this manner, more than 100 samples could be analyzed during a day using a refrigerated autosampler for overnight injection. The method was readily adapted to the determination of SarCNU in human plasma by simply decreasing the eluent flow-rate to 0.6 ml/min, whereby SarCNU and the I.S. eluted at approximately 5.8 and 9.1 min, respectively. Furthermore, the switching valve was not necessary for the analysis of human plasma samples. With a 50-microliter sample volume, the lowest concentration of SarCNU included in the plasma standard curves, 0.10 micrograms/ml, was quantified with a 7.8% R.S.D. (n = 27) over a 2 month period. Plasma standards, with concentrations of 0.26 to 5.1 micrograms/ml, exhibited R.S.D. values ranging from 1.3 to 4.7%. Thermospray-ionization MS detection was used to definitively establish the specificity of the method. The sensitivity of the assay was shown by application to be more than adequate for characterizing the plasma pharmacokinetics of SarCNU in mice. PMID- 8704942 TI - Separation of testosterone metabolites in microsomal incubates using a new capillary electrophoresis assay. AB - A capillary electrophoresis method has been developed to separate the products of liver microsomal testosterone metabolism. The microsomal mixture undergoes liquid liquid extraction and pre-concentration, and then electrophoretic analysis takes less than 25 min including capillary conditioning steps. The development of the complex background electrolyte (Tris-HCl and borate buffers, sodium dodecyl sulfate, beta-cyclodextrin and ethanol) necessary for this separation is described. A z-type capillary flow cell is used to obtain adequate detection sensitivity. The proportion in which the metabolites are produced as determined by this method allows assignment of the relative activity of cytochrome P-450 enzymes in the microsomes. The technique is useful for comparison of activity in normal and abnormal hepatic microsomes. PMID- 8704943 TI - Sensitive assay for verapamil in plasma using gas-liquid chromatography with nitrogen-phosphorus detection. AB - A sensitive gas-chromatographic method for quantitative analysis of verapamil in human plasma is described. The method involves a single extraction procedure, followed by separation on a capillary column and detection with a nitrogen phosphorus detector. The detection limit, based upon an assayed plasma volume of 0.5 ml, is 2 ng/ml. The standard curve is linear in the concentration range of 2 to 1000 ng/ml. The recovery of verapamil by pentane-isopropanol extraction was found to be 95%. Zipeprol is used as the internal standard. No interference from drugs needed for the associated cancer therapy has been found. Serum verapamil concentrations are determined by this method in fourteen cancer patients undergoing treatment with adriamycin. PMID- 8704944 TI - Rapid and selective high-performance liquid chromatographic method for the determination of metronidazole and its active metabolite in human plasma, saliva and gastric juice. AB - A rapid and selective HPLC method has been developed for the separation and quantitation of metronidazole and its hydroxylated metabolite in human plasma, saliva and gastric juice. The assay requires a simple protein precipitation step prior to analysis and is selective, sensitive and reproducible. The limits of quantitation (0.5-ml sample) were at least 0.25 microgram/ml for metronidazole and 0.20 micrograms/ml for its hydroxy metabolite. A Hypersil ODS 5 micron (150 x 4.5 mm I.D.) column was used with a mobile phase of acetonitrile-aqueous 0.05 M potassium phosphate buffer (pH 7) containing 0.1 % triethylamine (10:90) delivered at a flow-rate of 1.0 ml/min. PMID- 8704945 TI - Simple high-performance liquid chromatographic method for the quantitation of 5 fluorouracil in human plasma. AB - A simple, rapid, specific and sensitive high-performance liquid chromatography method has been developed for quantitation of 5-fluorouracil (5-FU) in human plasma. The method involves deproteinization of a small sample volume of plasma (150 microliters) followed by HPLC on a cation-exchange resin column, Aminex HPX 87H (300 x 7.8 mm I.D.), preceded by a similar guard cartridge with UV detection at 265 nm. This method allows a good separation of 5-FU with a retention time of 24 min and a detection limit at 25 ng/ml. The calibration curve was linear from 25 to 2000 ng/ml. The coefficient of variation was < or = 4.4% for within-day reproducibility and < or = 9.5% for day-to-day reproducibility. PMID- 8704946 TI - Measurement of physiological concentrations of dapsone and its monoacetyl metabolite: a miniaturised assay for liquid or filter paper-absorbed samples. AB - A modification of existing HPLC assay methods is described for the measurement of dapsone and monoacetyldapsone in 50-microliter samples of plasma and whole blood. This method, in particular the use of small sample volumes dried onto filter paper strips, is applicable to multi-sample clinical and pharmacokinetic studies in children with malaria, who are often anaemic, and where sample volume must be kept to a minimum. Basified samples were extracted into 5 ml of ethyl acetate tert.-butylmethyl ether (1:1, v/v), chromatographed on a mu BondapaK C18, 10 micron column with water-acetonitrile-glacial acetic acid (81:17.5:5, v/v) containing 2 g/l l-octanesulphonic acid as the mobile phase and detected at 274 nm. PMID- 8704947 TI - Determination of the anti-platelet-activating factor BN-50727 and metabolites in human urine by high-performance liquid chromatography using solid-phase extraction. AB - A sensitive and selective HPLC solid-phase extraction procedure was developed for the determination of platelet-activating factor antagonist BN-50727 and its metabolites in human urine. The procedure consisted in a double solid-phase extraction of the urine samples on cyanopropyl and silica cartridges, followed by an automated solid-phase extraction of the drug and metabolites on CBA cartridges and posterior elution on-line to the chromatographic system for its separation. The method allowed quantitation in the concentration range 10-2400 ng/ml urine for both BN-50727 and the main metabolite, the O-demethylated BN-50727 product. The limit of quantitation for both compounds was 10 ng/ml. The inter-assay precision of the method, expressed as relative standard deviation, ranged from 1.9 to 4.5% for BN-50727 and from 2.5 to 9.0% for the metabolite. The accuracy, expressed as relative error, ranged from -2.4 to 4.2% and from 0.2 to 6.2%, respectively. This paper describes the validation of the analytical methodology for the determination of BN-50727 in human urine and also for its metabolites. The method has been used to follow the time course of BN-50727 and its metabolites in human urine after single-dose administration. PMID- 8704948 TI - Molecular genetic approaches for the study of virulence in both pathogenic bacteria and fungi. PMID- 8704949 TI - Molecular characterization of the levansucrase gene from the endophytic sugarcane bacterium Acetobacter diazotrophicus SRT4. AB - The Acetobacter diazotrophicus SRT4 gene encoding levansucrase (EC 2.4.1.10) (IsdA) was isolated from a genomic library. The nucleotide sequence of a 2.3 kb DNA fragment sufficient for complementation of a levansucrase-deficient mutant (obtained by EMS treatment) was determined. The IsdA gene (1751 bp) coded for a polypeptide of molecular mass 64.9 kDa with an isoelectric point of 5.2. The N terminal amino acid sequence of the extracellular levansucrase indicated the presence of a precursor protein with a putative signal sequence of 51 residues which is possibly cleaved in two successive steps. Expression of the IsdA gene from the lac promoter in Escherichia coli resulted in the production of a protein with levansucrase activity. The deduced amino acid sequence of the IsdA gene was 48% and 46% identical with the levansucrases from the Gram-negative bacteria Zymomonas mobilis and Erwinia amylovora, respectively, but only 28-31% identical with levansucrases from Gram-positive bacteria. Multiple alignments of published levansucrase sequences from Gram-negative and Gram-positive bacteria revealed eight conserved motifs. A comparison of the catalytic properties and the sequence of the A. diazotrophicus levansucrase with those of the Bacillus subtilis levansucrase suggested that one of these motifs may be involved in the specificity of the synthetized product. Disruption of the IsdA gene in the genome of A. diazotrophicus resulted in a mutant lacking both levansucrase activity and the ability to utilize sucrose as a carbon source, suggesting that levansucrase is the key enzyme in sucrose metabolism of A. diazotrophicus. PMID- 8704950 TI - Genetic transfer of amylovoran and stewartan synthesis between Erwinia amylovora and Erwinia stewartii. AB - DNA fragments with ams genes of Erwinia amylovora and cps genes of Erwinia stewartii were transferred to exopolysaccharide (EPS)-deficient mutants of the other species. The resulting EPSs were characterized by sensitivity to EPS dependent bacteriophages, staining with amylovoran-specific fluorescein isothiocyanate-labelled lectin and chemical techniques, such as determination of the sugar composition and methylation analysis in order to distinguish between amylovoran and stewartan. Degradation by the stewartan-dependent phage phi-K9 was used to detect stewartan production, and staining with a lectin from Abrus precatorius detected amylovoran capsules. The patterns of sugar linkages were determined by methylation analysis. Stewartan contained a significantly higher glucose to galactose ratio than amylovoran and produced a characteristic signal from 6-linked glucose residues. By these criteria, most E. stewartii cps mutants displayed exclusively amylovoran synthesis when complemented with the E. stewartii cps genes produced stewartan. The complementation to an EPS-positive phenotype may require most genes of the ams or the cps operon. An exception was an E. stewartii cpsK mutant that made predominantly stewartan when complemented with the ams cosmid. IR spectra showed that both amylovoran and stewartan were acylated when synthetized in E. amylovora, but not in E. stewartii. The amylovoran-producing E. stewartii merodiploids regained virulence to corn seedlings when mucoidy was restored by the ams cluster, but the stewartan producing E. amylovora ams-/cps+ strains were weakly virulent on pear slices and avirulent on apple seedlings. PMID- 8704951 TI - Application of a suite of 16S rRNA-specific oligonucleotide probes designed to investigate bacteria of the phylum cytophaga-flavobacter-bacteroides in the natural environment. AB - We designed a panel of four 16S rRNA-targeted oligonucleotide probes specific for bacteria of the phylum cytophaga-flavobacter-bacteroides (CFB). Probes CF319a and CF319b are targeted to members of the flavobacteria-cytophaga group and the genus Porphyromonas, whereas probe BAC303 has a target region characteristic for the genera Prevotella and Bacteroides within the bacteroides group. The probe FFE8b was developed for species-specific hybridizations with Flavobacterium ferrugineum. All probes were designed by computer-assisted sequence analysis and compared to all currently accessible 16S and 23S rRNA sequences. The oligonucleotides were further evaluated by whole-cell and non-radioactive dot blot hybridization against reference strains of the CFB phylum and other major lineages of Bacteria. The newly developed probes were used together with other higher-order probes to analyse the structure and community composition in complex environments. In activated sludge samples, members of the flavobacteria-cytophaga group were revealed by in situ hybridization as important constituents of sludge flocs and characteristic colonizers of filamentous bacteria. By application of fluorescent probe BAC303, members of the genera Bacteroides and Prevotella could be visualized without prior cultivation as an important part of the human faecal microflora. PMID- 8704952 TI - The frequency of chimeric molecules as a consequence of PCR co-amplification of 16S rRNA genes from different bacterial species. AB - Our understanding of microbial diversity is greatly hampered by the inability to culture as much as 99% of the microbial community in the biosphere. Development of methods for identification and determining microbial phylogenies based on gene sequences, and for recovering genes directly from diverse environmental samples has made it possible to study microbes without the need for cultivation. PCR techniques have revolutionized retrieval of conserved gene sequences. However, it is well known that co-amplification of homologous genes may generate chimeric sequences leading to descriptions of non-existent species. To quantify the frequency of chimeric sequences in PCR amplification of 16S rRNA genes, we chose several 16S rDNAs with known sequences and mixed them for PCR amplifications under various conditions. Using this model system, we detected 30% occurrence of chimeric sequences after 30 cycles of co-amplification of two nearly identical 16S rRNA genes. The frequency of chimera formation decreased to 12.9% and 14.7% for templates with 82% and 86% similarity, respectively. We also examined effects of the number of amplification cycles, length of elongation periods and presence of damaged DNA on chimera formation. The results should provide useful information for microbiologists who use PCR-based strategies to retrieve conserved genes from mixed genomes. PMID- 8704953 TI - Degradation of 2,4-dichlorophenoxyacetic acid by haloalkaliphilic bacteria. AB - Three 2,4-dichlorophenoxyacetic acid (2,4-D)-degrading bacterial isolates were obtained from the highly saline and alkaline Alkali Lake site in southwestern Oregon contaminated with 2,4-D production wastes. While similar in most respects, the three isolates differed significantly in 2,4-D degradation rates, with the most active strain, I-18, demonstrating an ability to degrade up to 3000 mg 2,4-D I-1 in 3 d. This strain was well adapted to the extreme environment from which it was isolated, growing optimally on 2,4-D at pH 8.4-9.4 and at sodium ion concentrations of 0.6-1.0 M. According to its optimum salt concentration and pH for growth, this isolate was a moderately halophilic, alkaliphilic bacterium. The 16S RNA gene sequence (303 nt) was identical for all three isolates and most closely resembled those of the moderately halophilic eubacteria of the family Halomonadaceae (91% identity). Biochemical and genetic examination revealed strain I-18 utilizes the same 2,4-D degradation pathway as most of the 2,4-D degrading bacteria from non-extreme environments. Hybridization data and comparison of the partial sequences of the tfdA gene from the Alkali Lake isolates with those of bacteria from non-extreme environments suggested a common genetic origin of the 2,4-D degradation pathway in the two groups of micro organisms. PMID- 8704954 TI - Ether-bond scission in the biodegradation of alcohol ethoxylate nonionic surfactants by Pseudomonas sp. strain SC25A. AB - Pseudomonas sp. strain SC25A, previously isolated for its ability to grow on alcohol ethoxylates (PEG dodecyl ethers) as sole source of carbon and energy, was shown to be capable of growth on the dodecyl ethers of mono-, di, tri- and octaethylene glycols. Comparative growth yields for this series of alcohol ethoxylate nonionic surfactants indicated that, whereas all of the carbon of monoethylene glycol dodecyl ether (MEGDE) was assimilable, only the alkyl chains were assimilated from the higher ethoxamers. These results are interpreted in terms of a primary biodegradation mechanism in which the scission of the dodecyl ether bond is the first step. In the case of MEGDE this step separates the dodecyl chain from a C2 fragment, both of which are readily assimilable; for the higher ethoxamers, the assimilable dodecyl chain is accompanied by an ether containing PEG derivative which would require further rounds of either scission before assimilation. Whole cells and cell extracts converted [1-14C]MEGDE initially and very rapidly to radiolabelled dodecanol. Disappearance of [14C]dodecaol was accompanied by production of [14C]dodecanal. [14C]Dodecanoic acid was present at relatively low concentrations throughout the incubation periods. [14C]Dodecan-1, 12-dioic acid was produced in significant quantities (up to 25% radiolabel), and the onset of its production coincided with the peak concentration of dodecanal, the disappearance of which mirrored the appearance of the dioic acid. Under anaerobic conditions in the presence of cell extracts, dodecanol (55% of radiolabel) and dodecanal (22%) accumulated rapidly from MEGDE, but there was little subsequent conversion to mono- or dicarboxylic acids. These results are interpreted in terms of a pathway initiated by dodecyl-ether cleavage to produce dodecanol, which is subsequently oxidized to dodecanal and dodecanoic acid. The formation of dodecan-1, 12-dioic acid, probably from dodecanal, may represent a means of harbouring carbon under non-growing conditions. PMID- 8704955 TI - Actin-related proteins in Anabaena spp. and Escherichia coli. AB - Actin has been described in all eukaryotic cells as the major microfilament cytoskeletal protein. Although prokaryotic cells do not have a cytoskeleton, proteins related to the latter have been found in different prokaryotic species. We have found prokaryotic actin-related proteins in the enterobacterium Escherichia coli and in the cyanobacteria Anabaena cylindrica and Anabaena variabilis. They were identified by the following criteria: (1) by cross-reaction with a fluorescent conjugated anti-actin (rat-brain) mAb by Western blot analysis (in total cellular extracts); (2) specific binding of acetone powder and soluble cellular extracts to DNase I; and (3) specific binding of cells and total cellular extracts to phalloidin. In E coli, specific binding of phalloidin labelled with rhodamine to cells was detected by spectrofluorometry. In total cellular extracts, three bands of 60, 43 and 35 kDa were weakly recognized by the mAb by Western blot analysis; this recognition increased when phalloidin was added to the extracts. Furthermore, three polypeptides of kDa were isolated by binding to DNase I, showing pI values of 6.7, 6.65 and 6.6, less acidic than all reported actin pI values. In A. cylindrica and A. variabilis, specific binding of phalloidin labelled with rhodamine to cells was also detected by spectrofluorometry. In total and soluble cellular extracts, the mAb recognized two bands of 45 and 40 kDa by Western blot analysis, but only the first was purified by binding to DNase I, and it showed three isoforms of pI values 6.8, 6.5 and 6.4. These results suggest the presence, in prokaryotes, of proteins with similar biochemical characteristics to eukaryotic actin. PMID- 8704956 TI - Reactions of the Escherichia coli flavohaemoglobin (Hmp) with NADH and near micromolar oxygen: oxygen affinity of NADH oxidase activity. AB - The soluble flavohaemoglobin (Hmp) of Escherichia coli, product of the hmp gene, contains haem B and FAD in a single polypeptide of molecular mass 44 kDa. The function of this protein (and of the similar proteins identified in several bacteria and yeast) is unknown, but the observation that the binding of oxygen to haem modulates the reduction level of FAD has suggested that Hmp could act as an oxygen sensor. Here, stopped-flow, rapid-scan spectroscopy has shown that the oxidized protein reacts rapidly with NADH to form an oxygenated species, even when efforts are made to reduce oxygen concentrations to sub-micromolar levels, suggesting a high affinity for this ligand. As is the case at high oxygen concentrations (130 microM), oxygenated species formation was kinetically and spectrally heterogeneous. Between 12 ms and 1 s after mixing, following transient formation of the deoxy form and its reaction with dioxygen, a steady-state level of the oxygenated species was attained. During the oxygenated steady state, the flavin remained largely oxidized, as observed previously at 130 microM oxygen. Hmp is an NADH oxidase; on exhaustion of oxygen by reduction (in < 10 s under these conditions), the oxygenated species disappeared to generate the deoxy Fe(II) haem, whereupon the flavin was reduced. The affinity for oxygen during NADH oxidation was measured by continuous dual-wavelength monitoring of the deoxygenation of oxymyoglobin. The Km for oxygen was 2.6 microM, much higher than the Km values determined, using the same method, for the membrane-bound terminal oxidases cytochromes bo' and bd. These results show that the oxidase activity of Hmp, but not necessarily oxygen binding, would be minimal at oxygen concentrations that limit terminal oxidase function. PMID- 8704957 TI - Demonstration of high-affinity Mn2+ uptake in Saccharomyces cerevisiae: specificity and kinetics. AB - The existence of multiple transport systems for Mn2+ in Saccharomyces cerevisiae has been demonstrated in this study. Mn2+ (supplied as MnCl2) was accumulated by S. cerevisiae at all Mn2+ concentrations examined (25 nM-1 mM) but a log-log plot of uptake rates and total amounts accumulated revealed the existence of at least two Mn(2+)-concentration-dependent transport systems. Over a low Mn2+ concentration range (25-1000 nM), high-affinity Mn2+ uptake occurred with a Km value of 0.3 microM, while transformation of kinetic data obtained over the concentration range 5-200 microM revealed another system with a Km of 62 microM. Meaningful kinetic analyses were not possible at high Mn2+ concentrations because of toxicity: only about 30% of cells remained viable after 30 min incubation with 1000 microM MnCl2. Release of K+ accompanied Mn2+ accumulation and this increased with increasing Mn2+ concentration. However, even in non-toxic Mn2+ concentration, the ratio of Mn2+ uptake to K+ release greatly exceeded electroneutral stoichiometric exchange. In 50 microM MnCl2, the ratio was 1:123 and this increased to 1:2670 in 1000 microM MnCl2, a toxic concentration. External Mg2+ was found to decrease Mn2+ accumulation at all concentrations examined, but to differing extents. Over the low Mn2+ concentration range (5-200 microM), Mg2+ competitively inhibited Mn2+ uptake with a half-maximal inhibitory concentration, Ki, of 5.5 microM Mg2+. However, even in the presence of a 50-fold excess of Mg2+, inhibition of Mn2+ uptake was of the order of 72% and it appears that the cellular requirement for Mn2+ could be maintained even in the presence of such a large excess of Mg2+. Over the high Mn2+ concentration range (5-200 microM), the Ki for Mg2+ was 25.2 microM. At low Mn2+ concentrations, Zn2+ and Co2+, but not Cd2+, inhibited Mn2+ uptake, which indicated that the high-affinity Mn2+ uptake system was of low specificity, while at higher Mn2+ concentrations, where the lower-affinity Mn2+ transport system operated, inhibition was less marked. However, competition studies with potentially toxic metal cations were complicated due to toxic effects, particularly noticeable at 50 microM Co2+ and Cd2+. PMID- 8704958 TI - Saccharomyces cerevisiae expressing bacterial polyhydroxybutyrate synthase produces poly-3-hydroxybutyrate. AB - The polyhydroxybutyrate (PHB) synthase gene of the bacterium Alcaligenes eutrophus was used to construct a yeast plasmid which enabled expression of the functional synthase enzyme in Saccharomyces cerevisiae. Cells transformed with the synthase plasmid accumulated up to 0.5% of cell dry weight as PHB, with accumulation occurring in the stationary phase of batch growth. The identity of PHB in recombinant yeast cells was confirmed with 1H-NMR spectra of chloroform extracted cell material. In addition, freeze-fracture electron microscopy revealed cytoplasmic granules exhibiting plastic deformations characteristic for PHB. GC results indicated a low background level of PHB in the wild-type strain, but intact polymer could not be detected by 1H-NMR. Formation of PHB in the recombinant strain implies the participation of native yeast enzymes in the synthesis of D-3-hydroxybutyryl-CoA (3-HB-CoA). Inhibition studies with cerulenin indicated that the fatty acid synthesis pathway is not involved in PHB precursor formation. Wild-type cell-free extracts showed D-3-HB-CoA dehydrogenase activity [150-200 nmol min-1 (mg protein)-1] and acetoacetyl-CoA thiolase activity [10-20 nmol min-1 (mg protein)-1], which together could synthesize monomer from acetyl CoA. PHB accumulation was simultaneous with ethanol production, suggesting that PHB can act as an alternate electron sink in fermentative metabolism. We propose that PHB synthesis in recombinant yeast is catalysed by native cytoplasmic acetoacetyl-CoA thiolase, a native beta-oxidation protein possessing D-3-HB-CoA dehydrogenase activity and heterologous PHB synthase. PMID- 8704959 TI - Novel pyoverdine biosynthesis gene(s) of Pseudomonas aeruginosa PAO. AB - Conjugational mobilization of a Pseudomonas aeruginosa PAO1 cosmid bank (in pMMB33) into a pyoverdine-deficient (pvd) mutant harbouring a mutation in the 47 min region of the chromosome yielded one clone which restored yellow-green pigmentation and fluorescence when grown on iron-deficient medium. The relevant pMMB33-derivative cosmid, pPYP17, contained a 15.1 kb insert which was subcloned into pKT240 as a 10.8 Sacl-CIal fragment conferring the same phenotype. This derivative, pPYP180, like pPYP17, also conferred an apparent wild-type phenotype on pvd mutants previously shown to map genetically in the 23 min region of the P. aeruginosa PAO chromosomes. Physical mapping indicated that the cloned DNA fragment is located at the 66-70 min region of the PAO chromosome, demonstrating that the restored apparent wild-type phenotype observed for the transconjugants was not the result of a true gene complementation. A gene interruption was obtained by replacing a 0.6 kb BgIll-BgIll region of pPYP180 necessary for the expression of the pigmentation/fluorescence phenotype, by a Hgr interposon (omega Hg). After conjugational transfer and introduction of the mutagenized fragment into the PAO1 chromosome by gene replacement, pyoverdine-deficient mutants were recovered, indicating that the fragment indeed contained at least one gene involved in pyoverdine synthesis. The yellow-green fluorescent compound produced by such cells harbouring plasmids pPYP17 or pPYP180 differed from pyoverdine in several aspects and was consequently named pseudoverdine. Although pseudoverdine was able to complex iron, it was unable to restore growth to pvd mutants in the presence of the iron chelator ethylenediamine di(o-hydroxyphenylacetic acid), or to mediate iron uptake into PAO1. Pseudoverdine lacked a peptide chain but possessed spectral properties similar to pyoverdine, suggesting that it was structurally related to the chromophore of the pyoverdine molecule. The recent structural determination of pseudoverdine as a coumarin derivative confirmed this view and sheds some light on the biosynthetic pathway of the pyoverdine chromophore. PMID- 8704960 TI - Acquisition of iron by the non-siderophore-producing Pseudomonas fragi. AB - The iron requirement, siderophore production and iron uptake mechanisms of the type strain Pseudomonas fragi ATCC 4973 and five P. fragi isolates from meat were analysed. The strains exhibited a high sensitivity to iron starvation: their growth was strongly inhibited in medium supplemented with the iron chelator ethylenediamine di(hydroxyphenylacetic acid) or in medium treated with 8 hydroxyquinoline to remove contaminating iron. No siderophores were detectable in the growth supernatants of iron-starved cells. Cross-feeding experiments in iron depleted medium showed, however, that the bacterial growth could be strongly stimulated by siderophores of foreign origin including desferriferrioxamine B, enterobactin and some pyoverdines. Moreover, all the strains were capable of efficiently using the iron sources present in their natural environment, i.e., transferrin, lactoferrin and haemoglobin. Iron starvation led to the specific production of supplementary outer-membrane proteins of apparent molecular mass ranging from 80 to 88 kDa. Furthermore, growth in the presence of exogenous siderophores resulted, in some strains, in the induction of siderophore-mediated iron uptake systems. For one strain the concomitant synthesis of an iron regulated, siderophore-inducible outer-membrane protein was observed. PMID- 8704961 TI - Cell density influences antibiotic biosynthesis in Streptomyces clavuligerus. AB - Production of cephamycin C and clavulanic acid by Streptomyces clavuligerus took place during the exponential phase of growth in a defined medium. Both antibiotic biosynthetic pathways were activated shortly after spore germination, but the timing and kinetics of activation were affected by inoculum density. Rapid activation was favoured by high inoculum density or by growth in medium conditioned by previous incubation of S. clavuligerus spores or mycelium. A heat resistant conditioning factor able to accelerate the acquisition of antibiotic biosynthetic capacity when added to low-density cultures was released in suspensions of spores in water. Conditioning factor was also obtained in suspensions of spores from different Streptomyces species or of Bacillus cells, indicating that the signal was not produced specifically by S. clavuligerus. Fractionation of conditioning factor showed that its effect was not due to a single molecule. The fractions contained amino acids (as free amino acids and oligopeptides) in amounts that roughly correlated with their respective conditioning power. Furthermore, the conditioning effect was reproduced by supplementing defined medium with amino acids and peptides in concentration that mimicked those found in conditioning factor. When individually tested at concentrations in the micromolar range, only some amino acids were able to stimulate antibiotic biosynthetic capacity. This stimulation ws also promoted by low concentrations (less than 1 microgram ml-1) of peptide mixtures obtained with different proteolytic enzymes. The results suggest that both amino acids and peptides are responsible for the effects of conditioning factor released by spores. Possible implications of intercellular signalling on activation of secondary metabolism are discussed. PMID- 8704963 TI - In vitro phagocytosis and survival of Streptococcus suis capsular type 2 inside murine macrophages. AB - In this study, data on phagocytosis of Streptococcus suis and its survival inside macrophages are presented. Mouse peritoneal macrophages were incubated in the presence of one of five different strains of S. suis capsular type 2: a virulent wild-type strain (1591), a non-capsulated non-virulent mutant strain (M2), a poorly capsulated non-virulent mutant strain (M42), a non-virulent capsulated strain (1330), and the wild-type reference (virulent) strain S735. Opsonized or non-opsonized bacteria were incubated with macrophages in vitro and samples were obtained after 1 and 3 h incubation. Phagocytosis as well as live and dead intracellular organisms were determined by acridine orange and crystal violet staining. After 1 h incubation, non-opsonized virulent and non-virulent capsulated bacteria were poorly phagocytosed (by less than 7% of the macrophages), whereas the non-capsulated non-virulent mutant strain was highly phagocytosed (by more than 68% of the macrophages). The M42 mutant strain was more phagocytosed than the capsulated strains but less than the non-capsulated M2 mutant strain (35%). In contrast, a higher percentage of live bacteria was observed inside macrophages for the capsulated strains (1591 and S735) than for the non- or poorly capsulated mutant strains (M2 and M42). Opsonization of bacteria with rabbit serum or heat-inactivated rabbit serum significantly increased phagocytosis. For every opsonized strain, after 3 h incubation, the percentage of live bacteria within macrophages was considerably lower than the corresponding non-opsonized strains. In conclusion, the capsule of S. suis type 2 appears to act as an important anti-phagocytic factor. However, virulent capsulated non-opsonized strains can be phagocytosed by mouse peritoneal macrophages within which they appear to survive for at least 3 h. Serum factors other than complement increase not only phagocytosis but also intracellular killing of S. suis of both capsulated and non-capsulated strains. PMID- 8704962 TI - Metabolism of glycoprotein-derived sialic acid and N-acetylglucosamine by Streptococcus oralis. AB - Nine strains of Streptococcus oralis, isolated from blood cultures of patients with infective endocarditis or from the oral cavity as part of the normal flora, were examined for their ability to elaborate sialidase (neuraminidase) and N acetylglucosaminidase, enzymes which are involved in the degradation of glycoproteins. Both glycosidases were induced when bacteria were grown in a minimal medium supplemented with porcine gastric mucin, a model glycoprotein, and repressed when growth occurred in the presence of glucose. Cell-free extracts mucin-grown cultures expressed elevated levels of N-acetylneuraminate pyruvate lyase (the first intracellular enzyme in the pathway of N-acetylneuraminate catabolism), N-acetylglucosamine (glcNAc)-6-phosphate deacetylase and glucosamine 6-phosphate deaminase (enzymes involved in the intracellular catabolism of GlcNAc 6-phosphate); activity of each of these intracellular enzymes was markedly repressed when bacteria were grown in media supplemented with alpha 1-acid glycoprotein, a major component of human plasma. Cells from these cultures expressed high levels of sialidase, N-acetylglucosaminidase, and the intracellular enzymes involved in the catabolism of N-acetyl-sugars released by action of these glycosidases. High-resolution 1H-NMR spectroscopy of spent culture supernatants revealed that sialic acid and GlcNAc residues of the molecularly mobile oligosaccharide side-chains of alpha 1-acid glycoprotein had been hydrolysed and the released sugars internalized by the bacteria. These data indicate that S. oralis has the ability to hydrolyse constituents of oligosaccharide side-chains of host-derived glycoproteins and to utilize simultaneously these released carbohydrates. The biochemical characteristics induced by the growth of S. oralis on glycoproteins may play a role in the survival and persistence of these bacteria at the infection site in vivo. PMID- 8704964 TI - Dynamic expression of cell-surface antigens probed with Candida albicans-specific monoclonal antibodies. AB - IgG hybridomas were produced with preferentially reacted with cell-surface antigens of either yeast cells or hyphae of Candida albicans. Four mAbs were used in an immunostaining procedure to follow the expression dynamics of these antigens in media supplemented with glucose or galactose. Yeast cell growth was analysed during the lag phase, the early- and late-exponential phases and the stationary phase, and mycelium formation was analysed between 0.5 and 24 h induction at 37 degrees C. It appears that yeast cell-surface antigens 5C11 and 2E11 are expressed throughout all phases of yeast cell growth as well as on young hyphae after up to 1 h induction. Longer hyphae only faintly react with these two mAbs as they switch to hyphal cell-surface antigens 2G8 and 4E1 after 3 h induction. The reactivity to mAbs 2G8 and 4E1 was induced after a 3 h temperature shift and was confined to the terminal third of growing mycelia. Growth and hyphae induction in galactose prolonged the reactivity of young hyphae with the two anti-yeast-cell mAbs, whereas the expression of surface antigens 2G8 and 2E11 appeared delayed and desynchronized on hyphae. Whereas a similar reactivity was found with ten ATCC strains of C. albicans, four clinical isolates had a unique pattern of reactivity. Immunoblot analyses of DTT extracts of cell-surface constituents indicated that the antigens were proteinaceous in nature and showed that yeast-cell antigens 5C11 and 2E11 are detected in four bands between 68 and 104 kDa, whereas mycelial antigens 4E1 and 2G8 are detected in 117 kDa and 104 kDa bands found in mycelial but not in yeast-cell extracts. Present data support the concept of a dynamic balance in the expression of phase-specific antigens in C. albicans. PMID- 8704965 TI - The role of the outer membrane in formaldehyde tolerance in Escherichia coli VU3695 and Halomonas sp. MAC. AB - To investigate the mechanism of formaldehyde tolerance in Gram-negative bacteria, two formaldehyde-tolerant strains, Escherichia coli VU3695 and Halomonas sp. MAC (DSM 7328), and formaldehyde-sensitive revertants obtained by ethidium bromide or novobiocin treatment were studied. The presence of high levels of formaldehyde dehydrogenase activity alone proved insufficient to confer tolerance to high formaldehyde concentrations, as shown by high activity displayed by formaldehyde sensitive revertants of Halomonas MAC. Moreover, formaldehyde-tolerant strains also proved to be tolerant to high concentrations of acetaldehyde and glutaraldehyde, which are not oxidized by formaldehyde dehydrogenase. Treatment with sublethal concentrations of EDTA rendered the resistant strains highly sensitive to formaldehyde without affecting the activity of formaldehyde dehydrogenase. Comparison of the outer membrane proteins of formaldehyde resistant strains with those of their sensitive revertants showed the presence of at least one additional high molecular mass protein in the tolerant strains. It is concluded that formaldehyde tolerance in the bacteria studied depends on the composition and structure of the outer membrane. PMID- 8704966 TI - Transcription of the glnB and glnA genes in the photosynthetic bacterium Rhodospirillum rubrum. AB - The PII protein, encoded by glnB, has a central role in the control of nitrogen metabolism in nitrogen-fixing prokaryotes. The glnB gene of Rhodospirillum rubrum was isolated and sequenced. The deduced amino acid sequence had very high sequence identity to other PII proteins. The glnA gene, encoding glutamine synthetase, was located 135 bp downstream of glnB and was partially sequenced. glnB is cotranscribed with glnA from a promoter with high similarity to the sigma 54-dependent promoter consensus sequence. A putative sigma 70 promoter was also identified further upstream of glnB. Northern blotting analyses showed that in addition glnA is either transcribed from an unidentified promoter or, more likely, that the glnBA transcript is processed to give the glnA mRNA. The total level of the two transcripts was much higher in nitrogen-fixing cells than in ammonia-grown cells. PMID- 8704967 TI - Genes responsible for nisin synthesis, regulation and immunity form a regulon of two operons and are induced by nisin in Lactoccocus lactis N8. AB - Nisin is a small post-translationally modified lanthionine-containing peptide (lantibiotic) produced by certain Lactococcus lactis strains which has a high antimicrobial activity against several pathogenic Gram-positive bacteria. Northern blots and RT/PCR analysis of the nisin-producing strain N8 revealed that the nisZBTCIPRKFEG gene cluster, responsible for nisin biosynthesis, immunity and regulation, consists of two operons, nisZBTCIPRK and nisFEG. The promoter of the nisFEB operon was mapped. The -35 to -1 region upstream of the transcription start of the nisFEG promoter showed 73% identity with the corresponding region upstream of the nisA and nisZ gene. In contrast to earlier reports, nisin was found to be secreted during the early stages of growth was well as later in the growth cycle. The secreted nisin was adsorbed on the surface of the cells and was released to the medium during mid-exponential growth, when the pH in the medium fell below 5.5. In nisZB antisense and nisT deletion mutant strains constructed in this study the transcription of the nisin operons, nisin production and immunity were lost. Provision of external nisin restored the transcription of both operons in the mutant strains, showing that the operons are coordinately regulated by mature nisin. Nisin induction of the mutant strains also resulted in an increased amount of the NisI protein and an increase in the level of immunity. Induction using higher concentrations of nisin yielded a higher level of immunity. These results showed that the nisin promoters are under positive control in an autoregulatory manner and that antimicrobial peptides can also function as signal molecules. PMID- 8704968 TI - Regulation of bacterial methane oxidation: transcription of the soluble methane mono-oxygenase operon of Methylococcus capsulatus (Bath) is repressed by copper ions. AB - Methane is oxidized to methanol by the enzyme methane mono-oxygenase (MMO) in methanotrophic bacteria. In previous work, this multicomponent enzyme system has been extensively characterized at the biochemical and molecular level. Copper ions have been shown to irreversibly inhibit MMO activity in vivo and in vitro, but the effect of copper ions on transcription of the genes encoding the soluble (cytoplasmic) MMO (sMMO) has not previously been investigated. To examine more closely the regulation of bacterial methane oxidation and to determine the role of copper in this process, we have investigated transcriptional regulation of the sMMO gene cluster in the methanotrophic bacterium Methylococcus capsulatus (Bath). Using Northern blot analysis and primer extension experiments, it was shown that the six ORFs of the sMMO gene cluster are organized as an operon and the transcripts produced upon expression of this operon have been identified. The synthesis of these transcripts was under control of a single copper-regulated promoter, which is as yet not precisely defined. PMID- 8704969 TI - Promoters from Corynebacterium glutamicum: cloning, molecular analysis and search for a consensus motif. AB - Relatively limited information about promoter structures in Corynebacterium glutamicum has been available until now. With the aim of isolating and characterizing such transcription initiation signals, random Sau3A fragments of C. glutamicum chromosomal DNA and of the corynebacterial phage phi GA1 were cloned into the promoter probe vector pEKplCm and selected for promoter activity by chloramphenicol resistance of transformed C. glutamicum cells. The nucleotide sequence of ten chromosomal and three phage fragments was determined and the transcriptional start (TS) sites were localized by primer extension analyses. Additionally, the promoters of five previously isolated C. glutamicum genes were cloned and mapped. All of the isolated promoters were also functional in the heterologous host Escherichia coli. A comparative analysis of the newly characterized promoter sequences together with published promoters from C. glutamicum revealed conserved sequences centred about 35 bp (ttGcca) and 10 bp (TA.aaT) upstream of the TS site. The position of these motifs and the motifs themselves are comparable to the -35 and -10 promoter consensus sequences of other Gram-positive and Gram-negative bacteria, indicating that they represent transcription initiation signals in C. glutamicum. However, the C. glutamicum consensus hexamer of the -35 region is much less conserved than in E. coli, Bacillus, Lactobacillus and Streptococcus. PMID- 8704970 TI - Molecular characterization of adiY, a regulatory gene which affects expression of the biodegradative acid-induced arginine decarboxylase gene (adiA) of Escherichia coli. AB - A transcriptional regulator gene, designated adiY, was found downstream of the biodegradative arginine decarboxylase (adiA) gene (previously known as adi) of Escherichia coli. The arginine decarboxylase system is maximally induced under conditions of acidic pH, anaerobiosis and rich medium, and AdiY was found to increase the expression of adiA. The DNA sequence of adiY encodes a protein of 253 amino acids. Primer extension analysis defined the promoter. The amino acid sequence of AdiY showed homology to the XylS/AraC family of transcriptional regulators, which includes EnvY and AppY. Studies suggested that sequences required for acid induction were also necessary to observe the stimulation by AdiY. An examination of the substitution of AdiY, AppY and EnvY showed that these three proteins can, to some extent, stimulate the other systems. PMID- 8704971 TI - An abundant hydrophobin (ABH1) forms hydrophobic rodlet layers in Agaricus bisporus fruiting bodies. AB - The SDS-insoluble protein fraction of Agaricus bisporus fruiting bodies was solubilized with trifluoroacetic acid. On SDS-PAGE this fraction was found to contain one abundant protein with an apparent M(r) of 16 kDa. The N-terminal amino acid sequence of this protein was determined and RT-PCR used to isolate a cDNA clone which upon sequencing identified the protein as a typical class I hydrophobin (ABH1). The gene (ABH1) was isolated and sequenced, and a second hydrophobin gene (ABH2) was found about 2.5 kbp downstream of ABH1. Purified ABH1 self-assembled at hydrophobic-hydrophilic interfaces, producing the typical rodlet layer known from other hydrophobins. Similar rodlets were observed on the surface of the fruiting body, while immunological localization showed the hydrophobin to be particularly abundant at the outer surface of fruiting bodies, in the veil and in the core tissue of the stipe. Transcripts of ABH1 were found only in fruiting-body hyphae. The ABH1 hydrophobin is probably solely responsible for the hydrophobicity of the fruiting-body surface but may also line air channels within fruiting bodies. PMID- 8704972 TI - Unusual inositol triphosphate metabolism in yeast. PMID- 8704973 TI - 1995 Colworth Prize Lecture. The regulation of antibiotic production in Streptomyces coelicolor A3(2). PMID- 8704974 TI - A role for pabAB, a p-aminobenzoate synthase gene of Streptomyces venezuelae ISP5230, in chloramphenicol biosynthesis. AB - Mutagenesis of Streptomyces venezuelae ISP5230 and selection for P-aminobenzoic acid-dependent growth in the presence of sulfanilamide yielded pab mutants (VS519 and VS620) that continued to produce chloramphenicol (Cm), although with increased medium dependence. Transforming the mutants with pDQ102 or pDQ103, which carried a pab-complementing fragment from S. venezuelae ISP5230 in alternative orientations, restored uniformly high Cm production in VS620, but did not alter the medium dependence of Cm production in VS519. The cloned S. venezuelae DNA fragment was subcloned and trimmed to the minimum size conferring pab complementation. The resulting 2.8 kb BamHl-Sacl fragment was sequenced. Codon preference analysis showed one complete ORF encoding a polypeptide of 670 amino acids. Comparison of the deduced amino acid sequence with database proteins indicated that the N- and C-terminal regions resembled PabA and PabB, respectively, of numerous bacteria. The gene product showed overall sequence similarity to the product of a fused pabAB gene associated with secondary metabolism in Streptomyces griseus. Insertion of an apramycin resistance gene into pabAB cloned in a segregationally unstable vector and replacement of the S. venezuelae chromosomal pabAB with the disrupted copy lowered sulfanilamide resistance from 25 to 5 micrograms mL-1 and blocked Cm production. PMID- 8704975 TI - Gene expression in the cos region of the Streptomyces temperate actinophage phi C31. AB - A transcription map of a 5.12 kb region containing the cos ends of actinophage phi C31 was determined using RNA prepared from induced and uninduced cultures of the temperature-sensitive lysogen, Streptomyces coelicolor A3(2) (phi C31cts1). In induced cultures, RNA synthesis was detected only late in the lytic cycle. A late operon initiated downstream of the int gene on the right-hand end of the genome traversed the cos ends and extended at least 3.6 kb along the left-hand end. Shorter, possibly processed, mRNAs were also present. The map was superimposed on the DNA sequence of 2.8 kb of the region, part of which had been determined previously and part of which is presented here. The late-expressed transcripts contained a tRNA(Thr)-like gene and four ORFs (1, 2, 3 and 5) detected on the basis of codon bias. Analysis of the putative protein products showed that one of the ORFs could encode a lysis protein and at least one may be involved in DNA maturation. Transcription mapping of RNA from uninduced cultures demonstrated a 620 nt transcript, xRNA1, of ORF6. So far this is the only gene in phi C31 found to be expressed right to left with respect to the standard map of phi C31; its function during lysogenic growth could not be deduced from database searches. PMID- 8704976 TI - The requirement of tumour necrosis factor-alpha and interferon-gamma for the expression of protective immunity to secondary murine tularaemia depends on the size of the challenge inoculum. AB - The present study was conducted to determine the extent to which the cytokines tumour necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma) are required to protect against primary or secondary murine tularaemia caused by the live vaccine strain of the facultative intracellular bacterium Francisella tularensis. It is shown that non-immune mice treated with neutralizing monoclonal antibodies (mAbs) against TNF-alpha and IFN-gamma are rendered defenceless against otherwise sublethal intravenous inocula of the bacterium. Treatment with either of the anti-cytokine mAbs resulted in even a very small inoculum of 500 c.f.u. of the pathogen multiplying unrestrictedly in the livers, spleens and lungs of non-immune mice to rapidly reach lethal numbers. By contrast, Francisella-immune mice treated with either of the mAbs remained capable of resolving secondary infection with 50-fold larger inocula. However, the need for TNF-alpha and IFN-gamma for controlling secondary tularaemia became critical when challenge inocula exceeded 10(6) c.f.u. Overall, the results imply that different defence mechanisms operate to control primary versus secondary murine tularaemia. Additionally, they show that the need for TNF-alpha and IFN-gamma to combat secondary infection depends on the size of the challenge inoculum. PMID- 8704977 TI - A major T-cell-inducing cytosolic 23 kDa protein antigen of the vaccine candidate Mycobacterium habana is superoxide dismutase. AB - This study describes the purification and immunochemical characterization of a major 23 kDa cytosolic protein antigen of the vaccine candidate Mycobacterium habana (TMC 5135). The 23 kDa protein alone was salted out from the cytosol at an ammonium sulfate saturation of 80-95%. It represented about 1.5% of the total cytosolic protein, appeared glycosylated by staining with periodic acid/Schiff's reagent, and showed a pl of approximately 5.3. Its native molecular mass was determined as approximately 48 kDa, suggesting a homodimeric configuration. Immunoblotting with the WHO-IMMLEP/IMMTUB mAbs mc5041 and IT61 and activity staining after native PAGE established its identity as a mycobacterial superoxide dismutase (SOD) of the Fe/Mn type. The sequence of the 18 N-terminal amino acids, which also contained the binding site for mc5041, showed a close resemblance, not only with the reported deduced sequences of Mycobacterium leprae and Mycobacterium tuberculosis Fe/MnSODs, but also with human MnSOD. In order to study its immunopathological relevance, the protein was subjected to in vivo and in vitro assays for T cell activation. It induced, in a dose-related manner, skin delayed hypersensitivity in guinea-pigs and lymphocyte proliferation in BALB/c mice primed with M. habana. Most significantly, it also induced lymphocyte proliferative responses, in a manner analogous to M. Ieprae, in human subjects comprising tuberculoid leprosy patients and healthy contacts. PMID- 8704978 TI - Mechanisms regulating the transport of acetic acid in Saccharomyces cerevisiae. AB - Cells of the yeast Saccharomyces cerevisiae IGC 4072 grown in medium with acetic acid produced a mediated transport system for acetic acid that behaved as an electroneutral proton symport for the anionic form of the acid. The system could transport propionate and formate but not lactate and pyruvate. Uptake of labelled lactic acid was negligible, no mediated transport system activity for this acid being found. The acetate transporter was also found in cells grown in lactic acid or ethanol media, suggesting that the carrier did not require the presence of an external inducer. When cells were grown in lactic acid medium, uptake of labelled acetic acid, at pH 5.0, was biphasic and consistent with the presence of two distinct transport modes for the acid. One of these components corresponded to the acetate/proton symport, and the higher affinity system corresponded to a more general monocarboxylate carrier that could also transport lactate, pyruvate and propionate. Both systems were subject to repression by glucose, fructose, sucrose, maltose or galactose. In glucose-repressed cells, the undissociated form of the acids appeared to be the only one that could cross the plasma membrane, a diffusion mechanism being involved in the acid uptake. Under these growth conditions and when the extracellular pH was lower than that of the cytosol, accumulation of the acid could also be observed, it being a function of the delta pH. PMID- 8704979 TI - 'Fusel' alcohols induce hyphal-like extensions and pseudohyphal formation in yeasts. AB - At a concentration of 0.5% (v/v), isoamyl alcohol induced the formation of hyphal like extensions in haploid and diploid strains of Saccharomyces cerevisiae in liquid complex medium. These extensions, which develop via bud initiation and elongation, undergo DNA replication and nuclear division and appear similar in many respects to an aberrant form of the cell division cycle. However, in 0.25% (v/v) isoamyl alcohol, S. cerevisiae formed pseudohyphae. Other 'fusel' alcohols (which are the products of amino acid catabolism) also induced hyphal-like extensions in this yeast, with n-amyl alcohol being as equally effective as isoamyl alcohol. Isoamyl alcohol induced the formation of pseudohyphae in two species of Candida and both hyphal-like extensions and pseudohyphae in Brettanomyces anomalus, suggesting a close relationship or a common basis to the development of the two morphologies. PMID- 8704980 TI - Metabolic fluxes in chemostat cultures of Schizosaccharomyces pombe grown on mixtures of glucose and ethanol. AB - Simultaneous utilization of glucose and ethanol by the yeast Schizosaccharomyces pombe CBS 356 was studied in aerobic chemostat cultures. In glucose-limited cultures, respirofermentative metabolism occurred at growth rates above 0.16 h-1. Although Sch. pombe lacks a functional glyoxylate cycle and therefore cannot utilize ethanol as a sole carbon source, ethanol was co-consumed by glucose limited chemostat cultures. As a result, biomass yields increased, but not up to the theoretical value [0.92 g biomass (g glucose)-1] expected if all of the acetyl-CoA produced from glucose was instead synthesized from ethanol. When ethanol accounted for more than 30% of the substrate carbon in the mixed feed, it was incompletely utilized. In mixed-substrate cultures with a saturating ethanol fraction in the feed, the increase of the biomass yield as a result of ethanol consumption was highest at low dilution rates. This was not due to an increased specific rate of ethanol consumption at low growth rates; rather, the longer residence times at low dilution rates allowed Sch. pombe to utilize a larger fraction of the available ethanol, part of which was oxidized to acetate. Activities of gluconeogenic and glyoxylate-cycle enzymes were not detected in cell-free extracts of any of the cultures. Activities of acetaldehyde dehydrogenase and acetyl-CoA synthetase were low and of the same order of magnitude as the in vivo rates of acetate activation to acetyl-CoA. The results show that ethanol is a poor substrate for Sch. pombe, even as an auxiliary energy source. PMID- 8704981 TI - Determination of a 12 kb nucleotide sequence around the 76 degrees region of the Bacillus subtilis chromosome. AB - The nucleotide sequence of a 12361 bp DNA segment in the 76 degrees region of the Bacillus subtilis chromosome has been determined. Ten putative ORFs were identified. The deduced amino acid sequences of the products of two of them (glv 1 and glv-2) exhibited high similarity to those of glvG (6-phospho-beta glucosidase gene) and glvC [permease (the IIC domain) of the phosphotransferase system (PTS)], respectively, in the glv operon of Escherichia coli. The C terminal region of Glv-2 exhibited similarity to the entire region of GlvB (the IIB domain of PTS) of E. coli, suggesting fusion of the glvC and glvB genes in B. subtilis. glv-1, yfiA and glv-2 seem to form an operon of a phosphoenolpyruvate:sugar PTS, followed by a presumed four-membered operon of an ABC transport system. Moreover, a presumed sugar symporter and its regulatory genes were located in this region. PMID- 8704982 TI - Size and genomic location of the pMGA multigene family of Mycoplasma gallisepticum. AB - The pMGA multigene family encodes variant copies of the cell surface haemagglutinin of Mycoplasma gallisepticum. Quantitative Southern blotting, using an oligonucleotide probe complementary to a region conserved in the leader sequence of all known pMGA genes, was used to estimate the number of members of the family in the genome of seven strains of M. gallisepticum. The number of copies estimated to be present in the genome varied from 32 in strain F to 70 in strain R, indicating that the pMGA gene family may be second in size only to the tRNA family among prokaryotes. If all members of the pMGA family are of similar length to those which have been characterized, a minimum of 79 kb (7.7%) of the genome of strain S6, 82 kb (8.2%) of PG31 and 168 kb (16%) of the genome of strain R is dedicated to encoding variants of the same haemagglutinin. The GAA repeat motif identified in the intergenic region between all characterized pMGA genes appeared to be a feature common to most, if not all, pMGA genes, and furthermore probably exclusive to them. The genomic locations of members of the pMGA family were determined by PFGE and Southern blot hybridization of M. gallisepticum strain S6. The hybridizing regions were localized to four separate regions on the chromosome. The pMGA genes are likely to be predominantly arranged as tandem repeats within these regions, similar to the restricted regions for which the genomic sequence has been determined. PMID- 8704983 TI - Analysis of the sakacin P gene cluster from Lactobacillus sake Lb674 and its expression in sakacin-negative Lb. sake strains. AB - Sakacin P is a small, heat-stable, ribosomally synthesized peptide produced by certain strains of Lactobacillus sake. It inhibits the growth of several Gram positive bacteria, including Listeria monocytogenes. A 7.6 kb chromosomal DNA fragment from Lb. sake Lb674 encompassing all genes responsible for sakacin P production and immunity was sequenced and introduced into Lb. sake strains Lb790 and Lb706X which are bacteriocin-negative and sensitive to sakacin P. The transformants produced sakacin P in comparable amounts to the parental strain, Lb674. The sakacin P gene cluster comprised six consecutive genes: sppK, sppR, sppA, spiA, sppT and sppE, all transcribed in the same direction. The deduced proteins SppK and SppR resembled the histidine kinase and response regulator proteins of bacterial two-component signal transducing systems of the AgrB/AgrA type. The genes sppA and spiA encoded the sakacin P preprotein and the putative immunity protein, respectively. The predicted proteins SppT and SppE showed strong similarities to the proposed transport proteins of several other bacteriocins and to proteins implicated in the signal-sequence-independent export of Escherichia coli haemolysin A. Deletion and frameshift mutation analyses showed that sppK, sppT and sppE were essential for sakacin P production in Lb706X. The putative SpiA peptide was shown to be involved in immunity to sakacin P. Analogues of sppR and spiA were found on the chromosomes of Lb. sake Lb706X and Lb790, indicating the presence of an incomplete spp gene cluster in these strains. PMID- 8704984 TI - The Pseudomonas aeruginosa tonB gene encodes a novel TonB protein. AB - The Pseudomonas aeruginosa tonB gene was cloned by complementation of the tonB mutation of Pseudomonas putida strain TE516 (W. Bitter, J. Tommassen & P.J. Weisbeek, 1993, Mol Microbiol 7, 117-130). The gene was 1025 bp in length, capable of encoding a protein of 36860 Da. As with previously described TonB proteins, the P. aeruginosa TonB (TonBp.a.) was rich in Pro residues (18.1%) and contained Glu-Pro/Lys-Pro repeats. Unlike previously described TonB proteins, however, TonBp.a. lacked an N-terminal membrane anchor (signal) sequence and contained, instead, a predicted internal signal/anchor sequence, expected to yield an atypical N-terminal cytoplasmic domain in this protein. TonB proteins are essential components in iron-siderophore uptake in bacteria, apparently functioning as energy transducers in coupling the energized state of the cytoplasmic membrane to outer-membrane receptor function. As expected, tonB derivatives of P. aeruginosa were defective in siderophore-mediated iron acquisition. tonB gene expression was inducible by iron-limitation, consistent with the identification of a Fur consensus binding sequence upstream of the gene. TonBp.a. showed substantially greater similarity to the Escherichia coli TonB protein than the Pseudomonas putida protein (31% identity vs. 20% identity) and tonBp.a. was able to complement deficiencies in the acquisition of ferric enterobactin and vitamin B12, and sensitivity to phage phi 80 of an E. coli tonB strain. The larger size of TonBp.a. and its ability to function in both E. coli and P. putida make it a unique TonB protein whose characterization should enhance our understanding of TonB function in bacteria. PMID- 8704985 TI - Molecular characterization of a chromosomal region involved in the oxidation of acetyl-CoA to glyoxylate in the isocitrate-lyase-negative methylotroph Methylobacterium extorquens AM1. AB - A region on the Methylobacterium extorquens AM1 chromosome previously shown to complement a chemically induced mutant (PCT48) unable to convert acetyl-CoA into glyoxylate was characterized in detail in order to identify the gene(s) involved in the unknown pathway for acetyl-CoA oxidation. Six complete and two partial ORFs were identified by sequencing. Sequence comparisons suggested these might code for, respectively, a dehydrogenase of unknown specificity, a polypeptide of at least 15 kDa with unknown function, a coenzyme-B12-linked mutase, a catalase, an alcohol dehydrogenase (ADH) of unknown function, a polypeptide of 28 kDa, a ketol-acid reductoisomerase and a propionyl-CoA carboxylase (PCC). Insertion mutations were introduced into each ORF in order to determine their involvement in C1 and C2 metabolism. Mutations in three genes, encoding the mutase, ADH and PCC, resulted in a phenotype characteristic of mutants unable to oxidize acetyl CoA, i.e. they were C1-and C2-negative and their growth on these compounds was restored by the addition of glycolate or glyoxylate. Mutants in the genes thought to encode catalase and PCC were found to be deficient in the corresponding enzyme activity, confirming the identity of these genes, while physiological substrates for the mutase and ADH remain unidentified. This work, in which three new genes necessary for conversion of acetyl-CoA into glyoxylate were identified, is an intermediary step on the way to the solution of the unknown pathway for acetyl CoA oxidation in isocitrate-lyase-negative methylotrophs. PMID- 8704986 TI - Fur regulates the expression of iron-stress genes in the cyanobacterium Synechococcus sp. strain PCC 7942. AB - A homologue of the 'ferric uptake regulation' gene (fur) was isolated from the cyanobacterium Synechococcus sp. strain PCC 7942 by an Escherichia coli-based 'in vivo repression assay'. The assay uses a reporter-gene construct containing the promoter region of the iron-regulated cyanobacterial gene isiA, fused to the coding region for chloramphenicol acetyltransferase. The isolated gene codes for a protein that has 41% sequence similarity (36% identity) to Fur from E. coli and contains the putative iron-binding motif found in the Fur proteins of purple bacteria. No significant similarity was found to the DxtR repressor that regulates the expression of toxin and siderophore production in Gram-positive bacteria. Insertional mutagenesis of the cloned cyanobacterial fur gene led to the creation of heteroallelic mutants that showed iron-deficiency symptoms in iron-replete medium, including the constitutive production of flavodoxin and of hydroxamate siderophores. Failure to eliminate wild-type copies of the fur gene from the polyploid genome of Synechococcus 7942 implies that in this cyanobacterium Fur may have essential functions in addition to the regulation of genes involved in iron scavenging or photosynthetic electron transport. PMID- 8704988 TI - Proline is biosynthesized from arginine in Staphylococcus aureus. AB - Staphylococcus aureus NCTC 8325 exhibited a long lag phase (11 h) when inoculated into defined medium lacking proline, that could be shortened by increasing the concentration of arginine in the medium, or by supplying ornithine. Radioactivity from L-[14C]arginine, but not L-[14C]glutamate was incorporated into a spot with the chromatographic mobility of [14C]proline in the pool metabolites fraction. Selection for transposon Tn917-lacZ mutants impaired in arginine catabolism yielded four proline auxotrophs. Enzyme assays and precursor feeding experiments suggested that the major pathway for proline biosynthesis in S. aureus was from arginine via ornithine and delta'-pyrroline 5-carboxylate, rather than from glutamate. Strain 8325 Pro+, a proline prototrophic variant obtained by cultivation of 8325 in the absence of proline, accumulated L-[14C]arginine from the medium at about eight times the rate of strain 8325, suggesting its response to proline starvation was to increase arginine uptake. PMID- 8704987 TI - The QUTA activator and QUTR repressor proteins of Aspergillus nidulans interact to regulate transcription of the quinate utilization pathway genese. AB - Genetic evidence suggests that the activity of the native QUTA transcription activator protein is negated by the action of the QUTR transcription repressor protein. When Aspergillus nidulans was transformed with plasmids containing the wild-type qutA gene, transformants that constitutively expressed the quinate pathway enzymes were isolated. The constitutive phenotype of these transformants was associated with an increased copy number of the transforming qutA gene and elevated qutA mRNA levels. Conversely, when A. nidulans was transformed with plasmids containing the qutR gene under the control of the constitutive pgk promoter, transformants with a super-repressed phenotype (unable to utilize quinate as a carbon source) were isolated. The super-repressed phenotype of these transformants was associated with an increased copy number of the transforming qutR gene and elevated qutR mRNA levels. These copy-number-dependent phenotypes argue that the levels of the QUTA and QUTR proteins were elevated in the high copy-number transformants. When diploid strains were formed by combining haploid strains that contained high copy numbers of either the qutA gene (constitutive phenotype) or the qutR gene (super-repressing; non-inducible phenotype), the resulting diploid phenotype was one of quinate-inducible production of the quinate pathway enzymes, in a manner similar to wild-type. The simplest interpretation of these observations is that the QUTR repressor protein mediates its repressing activity through a direct interaction with the QUTA activator protein. Other possible interpretations are discussed in the text. Experiments in which truncated versions of the QUTA protein were produced in the presence of a wild-type QUTA protein indicate that the QUTR repressor protein recognizes and binds to the C-terminal half of the QUTA activator protein. PMID- 8704989 TI - An immunochemical study of serological cross-reaction between lipopolysaccharides from Vibrio cholerae O22 and O139. AB - A comparative chemical and serological study of the LPS of Vibrio cholerae O139 and O22 was performed. Chemical analysis revealed that the sugar composition of the LPS of strain O22 was quite similar to that of O139 LPS. Each contained D glucose, L-glycero-D-manno-heptose, colitose (3,6-dideoxy-L-galactose), D fructose, D-glucosamine, D-quinovosamine and D-galacturonic acid. The O-antigenic relationship between the two strains was analysed by passive haemolysis (PH) and passive haemolysis inhibition (PHI) tests with the respective LPS being used as antigens to sensitize sheep red blood cells (SRBC) and, in the latter case, as inhibitors in a PH system that consisted of LPS-sensitized SRBC, guinea-pig complement and anti-O139 or anti-O22 antiserum, both unabsorbed and absorbed with the heterologous antigen. In the PH experiment, unabsorbed anti-O139 antiserum had haemolytic titres of 66,000 and 22,000 against O139 LPS- and O22 LPS sensitized SRBC, respectively; unabsorbed anti-O22 antiserum had haemolytic titres of 900 and 13,000, respectively. Thus, the anti-O139 antiserum contained an antibody that reacted with a heterologous O22 antigen at a high titre (22,000) and this antibody was completely removed from anti-O139 antiserum with the O22 antigen. The anti-O22 antiserum contained an antibody that reacted with the heterologous O139 antigen at a low titre (900) and this antibody was completely removed from anti-O22 antiserum with the O139 antigen. In PHI tests O139 LPS and O22 LPS each strongly inhibited (the ID50 of LPS ranged from 0.03 to 0.14 microgram ml-1) the heterologous haemolytic systems of both O139 LPS-sensitized SRBC/anti-O22 antiserum and O22 LPS-sensitized SRBC/anti-O139 antiserm, which are substantially equivalent to the common antigen factor in the O139 LPS-sensitized SRBC/anti-O22 antiserum system and the common antigen factor in the O22 LPS sensitized SRBC/anti-O139 antiserum system, respectively. The results indicated that the O antigen of O139 is closely related to that of O22 in an a,b-a,c type of relationship where a is common antigenic factor, b is an O139-specific antigenic factor and c is an O22-specific antigenic factor. PMID- 8704990 TI - Expression of periplasmic alpha-amylase of Xanthomonas campestris K-11151 in Escherichia coli and its action on maltose. AB - A gene encoding the periplasmic alpha-amylase of Xanthomonas campestris K-11151 was cloned into Escherichia coli using pUC19 as a vector. An ORF of 1578 bp was deduced to be the amylase structural gene. The primary structure of the enzyme had little identity with other alpha-amylases, except with the enzyme from Bacillus megaterium. The enzyme was expressed in E. coli from the lac promoter of pUC19 and was found to be transported to the periplasmic space. The expressed enzyme showed the same thermal stability, optimum temperature and substrate specificity as the enzyme from X. campestris. The enzyme formed maltotetraose, but not 6(1)- nor 6(2)-maltosyl-maltose, from maltose by the reverse reaction, and the tetraose was then hydrolysed to maltotriose and glucose. The addition of maltotriose enhanced the production of glucose from maltose. In addition, maltose was formed by the condensation of glucose by the enzyme. Thus, the periplasmic alpha-amylase of X. campestris was shown to produce glucose from maltose by hydrolysing maltotetraose and possibly higher maltooligosaccharides, which were the products of a condensation reaction, as a major pathway, and by direct hydrolysis of maltose as a minor pathway. PMID- 8704991 TI - Extracellular and surface-exposed polysaccharides of non-tuberculous mycobacteria. AB - We studied the outermost constituents of the cell envelopes, which are involved in the interaction between the bacilli and the host cells, of five pathogenic and non-pathogenic mycobacterial species for comparison with those we have previously characterized from M. tuberculosis. The extracellular materials (ECMs) were isolated by ethanol precipitation and compared to the surface-exposed materials (SXMs) extracted by mechanical means. The materials from both sources were composed almost exclusively of polysaccharides and proteins. Two groups of mycobacteria were clearly distinguishable. The first group comprised the pathogenic species M. kansasii which produced large amounts of ECM, the glycosyl composition of which was similar to that of the SXM. The second group comprised M. avium and the non-pathogenic strains of M. gastri, M. phlei and M. smegmatis which produced small amounts of ECK This latter group could be subdivided into those which produced carbohydrate-rich ECM (M. avium and M. gastri) and those forming protein-rich ECM (M. phlei and M. smegmatis), a classification that correlated with the difference in the growth rate of the two subgroups. The glycosyl composition of the ECM of a given species was qualitatively similar to that of the SXM, except for M. avium and M. phlei whose SXM were devoid of arabinose. In addition to glucose, mannose and arabinose, xylose was detected in the hydrolysis products of the ECM and SXM of M. smegmatis, the SXM of M. phlei and the ECM of some batches of M. avium. The polysaccharide constituents of the ECM and SXM of the different mycobacteria were purified by anion-exchange and gel filtration chromatography; all were found to be neutral compounds devoid of acyl substituents. The extracellular polysaccharides consisted of high-molecular-mass glycogen-like glucans, arabinomannans and mannans, structurally similar to the corresponding substances previously characterized from the capsule of M. tuberculosis. The same types of polysaccharides were characterized from the SXM of all the strains, except M. avium and M. phlei which were devoid of arabinomannans. This study questions the unique and universal representation of the mycobacterial cell envelope and the existence of the so-called acidic polysaccharide-rich outer layer. PMID- 8704992 TI - Identification of a 29 kDa protein in the envelope of Mycobacterium smegmatis as a putative ferri-exochelin receptor. AB - Evidence of a direct association between ferri-exochelin, the major extracellular siderophore of Mycobacterium smegmatis, and a 29 kDa protein has been obtained by three separate methods. (1) Direct binding of 55Fe(III)-exochelin by the 29 kDa protein in an envelope preparation from iron-deficient cells was demonstrated by the extraction of a complex with the non-denaturing detergent CHAPS, and subsequent CHAPS-PAGE and autoradiography. (2) Affinity chromatography on a chemically synthesized ferri-exochelin-Sepharose 4B matrix led to the retention of the 29 kDa protein and a 25 kDa protein. The smaller protein was partially eluted with 1 mM ferri-exochelin although it did not form a stable complex with ferri-exochelin. The 29 kDa protein could not be eluted from the affinity matrix with 1 mM ferri-exochelin either alone or with 1 M NaCl. Only 2% (w/v) SDS could do this, but resulted in protein denaturation. (3) Incubation of 55Fe-exochelin with CHAPS-solubilized envelope proteins in free solution followed by ion exchange chromatography resolved three radioactive peaks; subsequent analysis by SDS-PAGE showed that the peak with the highest 55Fe-binding activity per unit protein contained both the 29 and 25 kDa proteins. A direct association was demonstrated between the 29 kDa protein and 55Fe-exochelin by gel filtration. The evidence suggests that the 29 kDa iron-regulated envelope protein of M. smegmatis is a ferri-exochelin-binding protein and that the 25 kDa protein, which corresponds in size to a previously reported iron-regulated envelope protein in this bacterium, may have a role in the formation or maintenance of this complex. Proteins extracted from the cell envelope of iron-deficient M. smegmatis with CHAPS were dialysed to remove the detergent, incorporated into liposome suspensions and then incubated with 55Fe(III)-exochelin. This increased the retention of 55Fe by 133-fold compared to proteins not placed in liposomes. Retention of 55Fe was dependent upon the protein loading of the liposomes. Gel filtration confirmed that the iron was retained by these vesicles and even after dialysis the majority of 55Fe was still retained by the vesicles. Re solubilization of the labelled proteo-liposomes in various detergents gave limited recovery of a ferri-exochelin-protein complex. Attempts to resolve this complex by Triton X-100 PAGE led to separation of the two entities. The complex was stable, however, in a CHAPS-PAGE system. PMID- 8704993 TI - Adaptive response of the archaeon Sulfolobus acidocaldarius BC65 to phosphate starvation. AB - The adaptive response of the archaeon Sulfolobus acidocaldarius BC65 to phosphate starvation was studied. When cells were subjected to phosphate limitation, their growth was affected. In addition, the levels of synthesis and/or the degree of phosphorylation of several proteins changed, as detected by two-dimensional nonequilibrium pH gradient electrophoresis of cells labelled in vivo with [35S]methionine and [35S]cysteine, or H3 32PO4. After another growth-restricting treatment, a heat shock, a general inhibition of protein synthesis was observed. Under phosphate starvation conditions, a 36 kDa protein became phosphorylated without its synthesis being significantly modified, suggesting a probable regulatory role during adaptation of the cell to the change in the external environment. In Southern blot analysis with specific probes from very conserved regions of the phoR and phoB genes from Escherichia coli, a positive hybridization with S. acidocaldarius BC65 chromosomal DNA fragments was found. This suggested the presence in S. acidocaldarius BC65 of genes related to the E. coli genes involved in the phosphate starvation response system. This appears to be the first evidence of the possible existence of a two-component sensory system in a micro-organism from the archaeal kingdom Crenarchaeota. PMID- 8704994 TI - Collagen-binding activity of Prevotella intermedia measured by a microtitre plate adherence assay. AB - The ability of Prevotella intermedia to bind type I collagen was investigated. A simple method in which bacterial cells were allowed to attach to collagen-coated microtitre plate wells was used to characterize the activity. All strains of P. intermedia tested, as well as those of the closely related species Prevotella nigrescens, showed a capacity to attach to the collagen film. Exponential-phase cultures of P. intermedia demonstrated a greater binding capacity than older cells. Attachment to the collagen film was inhibited by the presence of EDTA, type I and IV collagen, denatured collagen (gelatin), fibrinogen or fibronectin. Pretreatment of bacterial cells with heat (60 degrees C, 30 min) or proteinase K also inhibited the binding. The collagen-binding activity could be solubilized from the bacterial cell surface by incubation with Zwittergent 3-14, a zwitterionic detergent. The collagen-binding capacity of P. intermedia demonstrated in the present study represents a mechanism of colonization allowing these bacteria to attach to a tissue matrix. PMID- 8704995 TI - Utilization of transferrin and salmon serum as sources of iron by typical and atypical strains of Aeromonas salmonicida. AB - The ability of typical and atypical strains of Aeromonas salmonicida to utilize non-haem sources of protein-bound iron was evaluated. (i) In a plate bioassay, the suppression of growth imposed on typical and atypical A. salmonicida by addition of the high-affinity iron chelator ethylenediamine-di(o- hydroxyphenylacetic acid) (EDDA) to the growth medium was reversed by the addition of 30% or 90% iron-saturated bovine or human transferrin (Tf) or lactoferrin (Lf) to the growth medium. (ii) The mechanism of obtaining iron from Tf was investigated by the addition of bovine Tf contained within a dialysis bag. The reversal of iron-restricted growth suppression differed between the strains in that the atypical strains were unable to utilize Tf contained within a dialysis bag while the typical strains were able to do so. This suggested a siderophore-mediated uptake of iron from Tf by the typical strains, which are known to produce siderophores while atypical strains do not. (iii) A solid-phase binding assay using horseradish-peroxidase-conjugated or biotinylated Tf or Lf failed to detect Tf/Lf-binding activity using whole typical or atypical cells. (iv) When atypical extracellular products (ECP) plus bovine Tf or salmon serum were enclosed in a dialysis bag, diffusible products were released which could reverse the EDDA-imposed growth suppression of an atypical strain. This reversal was negated by inhibition of the ECP metalloprotease with EDTA. (v) Purified 70 kDa serine protease of a typical strain was able to digest bovine Tf to low molecular mass fragments as observed in SDS-PAGE. These results indicate that typical and atypical strains of A. salmonicida differ in their mechanism of utilization of non-haem protein- bound sources of iron. Typical strains utilize Tf via a siderophore-mediated mechanism and are also able to digest Tf with the extracellular serine protease. Atypical strains utilize Tf by a siderophore independent mechanism probably involving the proteolytic degradation of Tf by the extracellular metalloprotease. PMID- 8704996 TI - The ribosome-inactivating protein restrictocin deters insect feeding on Aspergillus restrictus. AB - The fungus-feeding beetle, Carpophilus freemani, consumed equal quantities of young mycelia, fewer phialides bearing mature spores and much fewer phialides bearing developing spores of Aspergillus restrictus compared to those of Aspergillus nidulans when tested in diet choice assays. The degree to which specific fungal structures were consumed was inversely related to the localization of high levels of restrictocin, a ribosome-inactivating protein, to those structures. Pure restrictocin added to the insect diet at 1000 p.p.m. killed 38.5% of C. freemani larvae and 62.5% of Spodoptera frugiperda larvae in 48 h, but did not affect C. freemani adults or Helicoverpa zea larvae over the same interval. In diet choice assays, 1000 p.p.m. of restrictocin deterred feeding by adult C. freemani and Sitophilus zeamais compared to control diets. Thus, restrictocin production and localization may have a natural defensive role against insect feeding at times critical to spore formation by A. restrictus, and may have potential as an insect control agent. PMID- 8704998 TI - Postoperative respiratory complications. PMID- 8704997 TI - A putative cyclic peptide efflux pump encoded by the TOXA gene of the plant pathogenic fungus Cochliobolus carbonum. AB - Race 1 isolates of Cochliobolus carbonum are pathogenic on certain maize lines due to production of a host-selective cyclic tetrapeptide, HC-toxin. Flanking HTS1, which encodes the central enzyme in HC-toxin biosynthesis, a gene was identified and named TOXA. Like HTS1, TOXA occurred only in isolates of the fungus that make HC-toxin and was present as two linked copies in most toxin producing isolates. HTS1 and TOXA were transcribed in the opposite orientation and their transcriptional start sites were 386 bp apart. The predicted product of TOXA was a 58 kDa hydrophobic protein with 10-13 membrane-spanning regions. The sequence was highly similar to several members of the major facilitator superfamily that confer resistance to tetracycline, methylenomycin, and other antibiotics. Although it was possible to mutate one copy or the other of TOXA by targeted gene disruption, numerous attempts to disrupt both copies in a single strain were unsuccessful, suggesting that TOXA is an essential gene in strains that synthesize HC-toxin. On the basis of its presence only in HC-toxin-producing strains, its proximity to HTS1 and its predicted amino acid sequence, we propose that TOXA encodes an HC-toxin efflux pump which contributes to self-protection against HC-toxin and/or the secretion of HC-toxin into the extracellular milieu. PMID- 8704999 TI - Issues in asthma self-management. PMID- 8705000 TI - Communicating with temporarily nonvocal patients. PMID- 8705001 TI - Treatment for emphysema: an overview of lung volume reduction surgery. PMID- 8705002 TI - RNS president's message: crisis in health care: danger or opportunity for respiratory nursing? PMID- 8705004 TI - How do cellular telephones interfere with hospital equipment, and how can this be prevented? PMID- 8705003 TI - A mother's smoking behavior during pregnancy affects the fetus. PMID- 8705005 TI - A report on a respiratory nurse project in England. PMID- 8705006 TI - Follow-up visits for oxygen therapy patients. PMID- 8705007 TI - Education about tracheostomy care. PMID- 8705008 TI - Detection of specific antibodies in immune complexes of farmer's lung patients. AB - The authors examined 23 precipitin-positive symptomatic patients with Farmer's Lung(FL) and compared them to different groups of exposed asymptomatic precipitin positive(EAPP) and precipitin- negative(EAPN) farmers. The sera were tested using several techniques (i.e., immunodiffusion and ELISA for specific antibodies; polyethylene glycol [PEG] for circulating immune complexes [CIC]) in an attempt to find an in vitro test correlated with the disease which could also provide an insight into the pathogenic mechanisms of Farmer's Lung. Circulating immune complexes formed by IgG were significantly higher in Farmer's Lung patients than in EAPP subjects. In polyethlyene glycol precipitates from Farmer's Lung patients, specific antibodies found by ELISA correlated well with serum positivity, but they were not found in EAPP subjects. The possibility that the circulating immune complexes found were Ig aggregates was ruled out, as was the possibility that the antibodies found in the polyethylene glycol precipitate were also due to an unspecific link. The authors suggest that the circulating immune complexes of Farmer's Lung patients contain specific specific antibodies and that since their composition is different in EAPP subjects, these circulating immune complexes may play a role in the pathogenesis of the disease. PMID- 8705009 TI - Relationship between interferon-gamma, interleukin-4 and IgE production of lymphocytes from hen's egg-sensitive patients. AB - We measured in vitro interferon-gamma, interleukin-4 and IgE production of peripheral blood mononuclear cells stimulated with ovalbumin. Interferon-gamma in culture supernatants of ovalbumin-stimulated peripheral blood mononuclear cells from hen's egg- sensitive patients with atopic dermatitis was significantly higher than that of healthy children or hen's egg-sensitive patients with immediate symptoms. Furthermore, in patients with atopic dermatitis who were sensitive to hen's egg and patients with immediate symptoms, there was an inverse relationship between interferon-gamma and IgE production (r = -0.535, p <0.05), and significant correlation was found between interleukin-4 and IgE production (r = 0.802, p <0.05). For the above reasons, IgE synthesis of ovalbumin-stimulated peripheral blood mononuclear cells may be suppressed by interferon-gamma in patients with atopic dermatitis. In contrast, in patients with immediate symptoms, interleukin-4 may play a major role in the pathogenesis of increased IgE production. PMID- 8705010 TI - Characteristics of mold allergy. AB - The sensitization of three common species of molds (C. herbarum, A. alternaria, A. fumigatus) and other common inhalation allergens was studied by SPT (skin prick testing) and RAST in 677 asthmatic patients. Ten percent of patients had an SPT with a mean diameter of at least 2 mm, and 5% had a positive RAST (>0.35 PRU/l) to at least one mold species. A positive RAST was most strongly associated with a positive RAST to animals (odds ratio 9.5) and less positively with a positive RAST to house dust mites (odds ratio 3.9) and storage mites (odds ratio 1.8). Sensitization to mites was significantly associated with indices of damp dwellings, whereas sensitization to molds was not. This study suggests that sensitization to molds mostly occurs in patients with a high potential for being sensitized to common inhalation allergens and that sensitization to molds is not related to damp dwellings. PMID- 8705011 TI - Study of HLA-DQA1 alleles in celiac children. AB - The familial incidence of celiac disease (CD) confirms its genetic basis, although acquired factors are also involved. Many authors have reported a linkage between celiac disease and HLA antigens, but there are differences which depend on geographical areas, and nowadays the study must be done at the genetic level. Thirty-eight celiac children and 52 normal controls were included in this study. All individuals were chosen from the Castilla and Leon area. We used the reverse ?dot block? technique, using sequence-specific oligonucleotide DNA probes (Cetus, USA) to determine the HLA-DQA1 alleles in DNA samples previously amplified by PCR (polymerase chain reaction). The different frequency of alleles in patients and controls was assessed by 3 statistical tests: chi square (chi(2)), relative risk (RR) and etiologic fraction (EF). A very high frequency of DQA1*0201 (chi(2):p <0.0001) and DQA1*0501 (chi(2): p <0.0001) alleles was observed in patients; all but one (97%) had the DQA1*0501 allele vs. 40% of controls (RR: 37.00; EF: 0.955). The DQA1*0201 allele also had a high prevalence in celiacs (58%)(RR: 1.375: EF:0.438). The DQA1*01 allele was only found in 10.5% of patients compared to 79% of controls (chi(2): p <0.0001) and the DQA1*03 allele was also decreased in celiacs. There was only one celiac girl without the DQA1*0501 allele. She had no other clinical or serological differences, as compared to the other patients. In the study of allele subtypes, among the DQA1*01 allele, 50% of patients were positive for DQA1*101 and the remaining 50% had DQA1*0102, but none of the individuals were positive for DQA1*0103. Among normal controls, 32 individuals (61.5%) expressed the DQA1*0102 subtype, 15 (28.9%) the DQA1*0101 subtype and 5 (9.6%) the DQA1*0103 subtype. All positive cases for DQA1-*05 belong to the DQA1* 0501 subtype, in both celiac and control groups. There were 10 possible combinations of HLA-DQA1 genes, but we found a very unequal distribution in both celiacs and controls. Only 4 genotypes were found in patients (DQA1*01/1*0501, DQA1* 0201/1*0501, DQA1*03/1*0501 and DQA1*0501/1*0501) and 8 in controls. The DQA1*0201/1*-0501 genotype was the most discordant, being positive in 55.3% of patients vs. 3.8% of controls (chi2:p < 0.001; RR: 1.235; EF:0.534). None of the 90 individuals studied expressed the DQA1*0311*03 genotype. The DQA1*0201/1*03 genotype was not shown by any control and by only 1 celiac patient. It is noteworthy that the DQA1*0201/1*0501 genotype was more frequent than the homozygous genotype DQA1*0501/1*0501. Our results do not suggest a dosage effect for the DQA1*0501 allele. The determination of the HLA-DQA1 gene is a helpful tool for the screening of individuals with a high risk of being celiacs. PMID- 8705012 TI - Evaluation of disease activity by laboratory tests in juvenile rheumatoid arthritis. AB - We studied 29 juvenile rheumatoid arthritis patients with polyarticular onset in order to detect the presence of disease activity by laboratory tests. Laboratory studies included hemoglobin, hematocrit, erythrocyte sedimentation rate (ESR), white blood cell (WBC) and platelet counts. We also determined the levels of IgG, IgA, IgM, C3, C4 and C reactive protein, as well as the presence of rheumatoid factor and antinuclear antibodies. There were 13 patients in the group with polyarticular disease activity, and 16 in the group with asymptomatic juvenile rheumatoid arthritis. Low hematocrit and hemoglobin values, abnormal ESR and elevated WBC and platelet counts occurred in polyarticular juvenile rheumatoid arthritis patients with active disease. Increased levels of IgA, C3 and C4 were also found in the exacerbation stage, as compared to the asymptomatic control group. No differences were found in rheumatoid factor and antinuclear antibodies between the active and inactive juvenile arthritis patients. Our data agreed with previous studies in that a single laboratory test cannot necessarily confirm juvenile arthritis activity, and they suggest that two or more abnormal parameters would be useful in assessing the flare-up of the disease. PMID- 8705013 TI - Chemotaxis of alveolar macrophages and neutrophils in response to microbial products derived from organic dust. AB - Mechanisms of chemotaxis of alveolar macrophages (AMs) and neutrophils (PMNs) in response to microbial products derived from organic dust were studied using the blindwell chemotaxis chamber technique. Seven different known etiological agents causing respiratory symptoms were used for experiments: cell extract and endotoxin from Pantoea agglomerans (synonyms: Erwinia herbicola, Enterobacter agglomerans), cell extracts from Thermoactinomyces vulgaris and Aspergillus fumigatus, protease from Bacillus thermoproteolyticus rokko and two preparations of glucans. These agents were evaluated for their ability to direct attraction of alveolar macrophages and neutrophils and stimulation of alveolar macrophages to release chemotactic factors for other alveolar macrophages and neutrophils. The microbial products were able to attract both alveolar macrophages and neutrophils directly in a dose-dependent manner, and the exposure of cultured alveolar macrophages to most agents stimulated chemotactic activity for for alveolar macrophages and neutrophils. The generation and release of this activity by alveolar macrophages may provide a mechanism for the initiation and amplification of inflammatory reactions in the lung after inhalation of organic dust. Results of these in vitro studies may be relevant to the pathogenesis of alveolitis in organic dust-induced lung diseases. PMID- 8705014 TI - Detection of undegraded beta-lactoglobulins and evaluation of the molecular weight of peptides in hydrolysate cow's milk formulae. AB - Hydrolysate formulae have been developed with the purpose of reducing the allergenicity of cow's milk proteins, thus providing a suitable formula for feeding babies with cow's milk allergy. More recently, hydrolysate formulae have also been used with babies at high risk of atopy in order to prevent cow's milk allergy. The aim of the present study was to investigate the presence of intact cow's milk proteins and the molecular weights of peptides in several batches of two extensively hydrolyzed formulae and two partially hydrolyzed formulae. The results show the presence of a significant amount of intact beta-lactoglobulin in one partially hydrolyzed formula and of peptides with high molecular weights (>16,900 D) in the two partially hydrolyzed formulae. In conclusion, the present study confirms that partially hydrolysed formulae contain a large proportion of peptides with high molecular weights: in addition, undegraded beta-lactoglobulin was detected in a partially hydrolyzed whey formula. These data strongly indicate that partially hydrolyzed formulae may be not only allergenic in an already sensitized individual, but also immunogenic in a predisposed baby. PMID- 8705015 TI - Toxocariasis: a cause of hyper IgE and eosinophilia. AB - We present a patient with elevation of total serum IgE, eosinophilia and Toxocara exoantigen specific IgG and IgE antibodies (detected by ELISA and RAST, respectively). We ruled out atopy and other parasitic diseases common in our habitat (e.g., Echinococcus granulosus, Ascaris lumbricoides, Schistosoma mansoni). The patient was asymptomatic over the four years of the study, and the levels of total serum IgE, eosinophils and Toxocara exoantigen specific IgG and IgE remained high. Although at present, immunodiagnostic tests have a high sensitivity and specificity for toxocariasis, their value is limited. This is illustrated by the fact that in our patient these techniques could not distinguish between past and present infection. PMID- 8705016 TI - Adverse reaction to pseudoephedrine. AB - A patient developed a scarlatina-like rash on two separate occasions after receiving a dose of pseudoephedrine. Patch tests with this substance and other structurally related substances (i.e. ephedrine, phenylephrine, and epinephrine) were negative. The oral test with pseudoephedrine provoked a new episode. It is difficult to clarify the exact mechanism of the described reaction; the nature of this eruption probably resembles many other drug-induced adverse reactions in which there is no certainty if mechanisms of type I or III are involved. PMID- 8705017 TI - Accountability! PMID- 8705018 TI - Whatever it takes as long as it takes--the victory of the Mercy nurses. PMID- 8705019 TI - Market driven healthcare & its impact on nurses. PMID- 8705020 TI - Where do we go from here? PMID- 8705021 TI - A "drinking alcohol" program that reduces health & safety problems at home & at work. PMID- 8705022 TI - I won't do that. Nurses as conscientious objectors. PMID- 8705023 TI - A glimmer of hope. PMID- 8705024 TI - Is nurse staffing adequate? The Institute of Medicines seeks answers. PMID- 8705025 TI - Nurse abuse management style ... can nurses fight back? PMID- 8705026 TI - "You're it"! PMID- 8705027 TI - "Come into my parlor," said the spider to the fly: a guide to working on hospital management committees. PMID- 8705028 TI - How to drive nurses crazy and out of the profession! PMID- 8705029 TI - Nursing and political action in the 20th century: from separation to fusion. PMID- 8705030 TI - Utilizing the "force field" analysis. PMID- 8705031 TI - Strike back! PMID- 8705032 TI - A nurses bad day. PMID- 8705033 TI - Walk with me. PMID- 8705034 TI - The view askew. PMID- 8705035 TI - "Can't you take a joke?" Sexual harassment in healthcare. PMID- 8705036 TI - The Regents College Nursing Program--problems & solutions. PMID- 8705037 TI - Revolution follows the breast cancer epidemic. One baby step forward. PMID- 8705038 TI - Nurses on the move. Rules for travel nursing. PMID- 8705039 TI - Nurse talk radio is on the air! PMID- 8705040 TI - Are nurseless hospitals coming! Not if I can help it! PMID- 8705042 TI - Helping your patients manage managed health care. PMID- 8705041 TI - New threat to U.S. healthcare hospitals without nurses. PMID- 8705043 TI - Express yourself at your own risk! PMID- 8705044 TI - Will revolutionary nurses signal the dawn of a new day? PMID- 8705045 TI - The terminology of patient-focused care: nouns as verbs, adjectives as nouns. PMID- 8705046 TI - Patient focused care: consultants, foundations, educational programs. PMID- 8705047 TI - Back to the dark ages of nursing. PMID- 8705048 TI - Flo's girl. Interview by Kate Rounds. PMID- 8705049 TI - Just a nurse. PMID- 8705050 TI - Joining the club: a nurse's personal experience with breast cancer. PMID- 8705051 TI - Let's make health research a #1 priority. PMID- 8705052 TI - Bertha. PMID- 8705053 TI - Where have all the nurses gone? PMID- 8705054 TI - Nurses and HIV. Beware the experts! PMID- 8705055 TI - Who needs a nurse? PMID- 8705056 TI - Workplace violence--nurses at risk. PMID- 8705057 TI - Decision making in unsafe practice situations. PMID- 8705058 TI - The legal response. PMID- 8705059 TI - Do nurses know their rights under the Family and Medical Leave Act? PMID- 8705060 TI - ER nurses fight stereotypes on TV--a win for all nurses. PMID- 8705061 TI - OR nurses don't fight stereotypes on TV--a loss for all nurses. PMID- 8705062 TI - R.N. conscientious objectors to infant circumcision: a model for nurse empowerment. PMID- 8705064 TI - The reality of nursing in a multi-ethnic National Health Service. PMID- 8705065 TI - Can the introduction of markets help solve the problems facing the National Health Service today? PMID- 8705063 TI - The role of nurses in management with the National Health Service in Scotland. AB - The introduction of an internal market in health care in 1991 was the most radical change to the National Health Service (NHS) since its inception. The consequent NHS changes and reforms have had a profound impact on nursing management and inevitably on nurses in management at a personal level. This paper reports on the initial findings of a study of 158 nurses in management within the NHS in Scotland and addresses their demographic features and their management training/education pattern. It was found that the majority of respondents (77.8%) were working in their current position less than 4 years. There was a significant relationship between marital status and gender and current position (P < 0.05). A total of 58.8% of women in senior positions were single, on the other hand there was no single male respondent at a senior manager position. In total, 37 nurses in management (23.4%) had a degree qualification and there was a relationship between having a degree, age and current position. Overall, approximately half of the respondents had a formal management training qualification and only 10.1% (16 subjects) had a degree level management education. However, a large proportion (65.8%) of nurses in management believe that degree level education in management is required to be able to perform their work satisfactorily. PMID- 8705066 TI - Satisfaction among ambulatory surgery patients in two hospitals in Iceland. AB - This study aims to investigate the level of satisfaction of care received among patients undergoing ambulatory surgery in two hospitals in Iceland, using the Patient Satisfaction Instrument (PSI). The PSI consists of 25 items, broken down into three sub-scales which measure the patients' attitude towards nursing care. The sample consisted of ambulatory patients undergoing orthopaedic, urological, hernia or varicose veins operations. The patients were 16 years or older and we enlisted 70 individuals from each hospital. Overall, the results show that patients are generally satisfied with the level of care they have received. The patients in group I are more satisfied than the patients in group II; however, only four out of the 25 items which were scored attracted a significance level of < 0.05. The patients perceived the clinical skills of the nurses as satisfactory but felt they did not receive enough information about their operation from the nurses. Although the patients were generally satisfied with the level of care received, they also identified instances where they felt that the level of care was inadequate; however, as results from other studies show, patients often experience difficulties articulating something negative about their nurses and the care they have received. PMID- 8705067 TI - Computers conquer hospital wards in Germany. AB - As a result of the Health Reform Act (1993) German hospitals have to prepare for fundamental changes in their accountability to the health insurance companies. For example, the introduction of diagnosis-related groups and competitive elements into the health care sector necessitate thorough recording of patient related costs by each individual hospital. Consequently, the work done by nurses is to be measured in a more refined manner in order to obtain a correct picture of the overall costs for a particular treatment. Computer technology is seen as a suitable tool to meet the challenges of this new development. The Agnes Karll Institute of Nursing Research (German Nurses' Association) conducted a study in the hospitals of Hesse, Germany, in order to investigate the present state of nursing informatics and related questions. Some results will be outlined in this paper. PMID- 8705068 TI - A 'new' psychological contract for nurses: some management implications. AB - Changes within the health services are raising a number of employment issues for nurses. The idea that a professional qualification and a job will lead to security of employment and career development is rapidly changing. These assumptions, the 'old' psychological contract, is giving way to new expectations from employers and employees; the emergence of a 'new' psychological contract. A psychological contract is an implicit agreement between employer and employee that each party will treat the other fairly. Such contracts are maintained by virtue of all parties wanting to seek agreement on issues where possible and to maintain trust. While such a contract is not a legally binding agreement it is nonetheless a binding understanding between people. Changes to this psychological contract can have important implications for individuals and their employer in terms of work and organizational commitment. This paper will discuss some of the issues surrounding psychological contracts and the impact of violating them. It will also discuss, from a management perspective, how psychological contracts develop between employer and employee, and how to form a 'new' psychological contract based upon mutual benefit and shared values. PMID- 8705069 TI - The role of the practice nurse in primary health care: managing and supervising the practice nurse resource. AB - This paper, by drawing on material from a qualitative study funded by the Department of Health and Welsh Office, examines the supervision and management of the practice nurse in general practice. In doing so it explores the perspectives of practice nurses, the GPs who employ them, representatives from FHSAs, those responsible for commissioning community nursing services, and managers of community nursing provider units. Within this context the paper explores the nature of the supervisory relationship between GP and practice nurse, and the wider role of the FHSA, commissioners of community nursing services and managers of community nursing provider units. PMID- 8705070 TI - Multidisciplinary teamwork--myth or reality? AB - The provision and delivery of health care has become increasingly complex in recent years due to technological innovation in medicine and increasing client expectations. The challenge for health care workers is to meet the purchasers' demands, one option may be to shift working practices from a functionalist approach to one of multidisciplinary teamwork. This paper explores one example of multidisciplinary teamwork and examines the processes which enhance and those that limit collaboration. PMID- 8705071 TI - Nurses in commissioning: are we helping to build a stronger team? PMID- 8705072 TI - Balancing skill mix--future paediatric health care provision. AB - Issues of "cost vs quality" and the needs for "pairs of hands" rather than "pair of eyes" has meant that nursing service managers are having to look closely at the skill mix of the staff they employ. This paper outlines the need to define the role of the nurse and the function of skill mix review. Arguments are discussed surrounding the issues of an all qualified nursing staff in a traditional hospital or the use of a generic "multiskilled" workforce within a "patient-focused" environment. The reasons for developing the role of the National Vocational Qualification (NVQ) Support Worker are examined and how they, and the Project 2000 trained staff nurse compare, against each other when looking for staff to employ. This paper looks at the paediatric field of health care in the light of the recommendations made by the 1991 Clothier Report to increase the number of specialist trained staff within that environment and what the future role of the children's nurse may be PMID- 8705073 TI - Purchasing post-qualifying professional education in the health care sector. AB - The National Health Service (NHS) is proposing to establish local consortia for educational contracting, with wider involvement and responsibility being devolved to service managers. This paper addresses the educational issues of purchasing post-qualifying professional education for a cross-section of NHS hospital professionals. The professions chosen for comparison were clinical staff and include nurses, doctors, and as an example of paramedical professions', physiotherapists. PMID- 8705074 TI - Innovation in nursing management: professional, management and methodological considerations. AB - There are many changes occurring in the National Health Service (NHS) at this time, not just to economic and funding policies, but also at the very heart of nursing delivery. The introduction of "managerialism" into the senior clinical grades of nursing, midwifery and other professional staff has characterized the past few years. Against this backdrop is the increasing belief that NHS organizations must find improved ways of delivering patient care and other services. This has inexorably led to the consideration of diffusing innovation into practice as a way to improve performance and competitiveness. While there have been a number of clinical attempts at understanding this process, there has been very little written about innovation from the perspective of the nurse (or midwifery) clinical manager. This paper discusses some of the issues surrounding management innovation including the planning of interventions aimed at introducing innovation, and some of the methodological difficulties of studying complex organizations. The wider issues of innovation, nursing management and professional staff are also discussed. PMID- 8705075 TI - Drug abuse, AIDS, and the coming crisis in long-term care. AB - As the 21st century approaches and the proportion of the US population over 65 years of age increases, it is expected that the demand for long-term care will expand dramatically. This expectation has been widely discussed. Less widely discussed is a potential for increase in the demand for long-term care resulting, not from the geriatric conditions with which the industry is already familiar, but from chronic and debilitating substance abuse-related diseases. In the United States, the incidence of severe cardiovascular and hepatological deterioration in younger patients has begun to increase, partly due to the increased scope of drug use and the increased variety of drugs used by individuals under the age of 35. HIV disease progression resembles these degenerative conditions in some important ways, and HIV infection is now often accompanied by substance abuse disorders. Thus, the care of HIV-infected patients can serve as a model for the impact on the long-term care industry that this new type of patient is likely to make. Using HIV as a particularly dramatic example, this paper discuss those changes in detail, and concludes with recommendations for successfully adapting to them. PMID- 8705076 TI - Listening to patients in the National Health Service: a selective review of literature on patients' views about outpatient services in British hospitals. AB - An increased emphasis on quality assurance in the National Health Service (NHS) became evident in the literature related to health following the Government's White Paper, "Working for Patients" (1989) and the Patients' Charter (1991). The latter included specific quality standards to which purchasers and providers of health care made a commitment. This review of published and un-published (grey) literature concentrates on patient views about outpatient services in British hospitals, with special emphasis on the role of nurses. Findings indicate central emphasis on easily measurable factors, such as waiting times in outpatient departments and comparative neglect of more qualitative aspects, which may be of greater importance to patients. Specific attention has been given to exploring the role of nurses working in the outpatient department, which appears to be under-researched, based on this selective literature review. PMID- 8705077 TI - Conscience: the professional and the personal. AB - Conscience is sometimes thought of as private, personal, individual, possibly religious, and not be questionned. This paper seeks to present and make plausible an account of conscience as social, possibly secular, and fallible (and hence corrigible). In the light of this account of professional conscience is offered and conclusions are drawn about the questions that should be answered in the event of a nurse making an appeal to conscience. PMID- 8705078 TI - Patient-controlled analgesia: a review of the literature. PMID- 8705079 TI - A model of clinical supervision for nurses undertaking short placements in mental health care settings. AB - This paper presents a model of supervision developed from the experience of supervising undergraduate general nursing students undertaking clinical experience in psychiatric wards. Supervision structures and methods are presented and the theoretical under-pinnings of the supervision model are discussed. Our experience of using the model over the past 5 years suggests that it has the potential to allow students to gain a good understanding of individual patients within a short time span and to engage with them as active collaborators in care. The supervision model can also assist students to develop the knowledge and insights needed to gain the respect of senior colleagues, and the confidence to challenge the professional conservatism that often characterizes psychiatric wards. PMID- 8705080 TI - Feminist approaches to change and nursing development. AB - In Britain, nursing development units (NDUs) have provided a focus for change and innovation in nursing since the early 1980s. This paper explores the approaches to change in these areas of practice. A review of both the management of change and feminist literature has been conducted. By identifying the similarities between the management of change in NDUs and feminism, it is suggested that feminist philosophies as demonstrated in NDUs may provide the key to successful and sustained, patient-focused changes in practice. PMID- 8705081 TI - Credit where credit's due--work-based learning in professional practice. AB - Professional education is constantly changing. It is important to value practice as an integral part of professional education. Work-based learning offers a vehicle for the linking of theoretical and clinical academic development. The tripartite agreement between employer, student and university can be beneficial to all parties. PMID- 8705082 TI - What do elderly people do in hospital? AB - Current health-care reforms have sharpened the focus on efficiency of bed usage; one useful method of exploring this concept is to determine how patients spend their time in hospital. This small study explored how 23 elderly patients spent their time on two elderly care wards. A time-sampling, patient-focused, structured observation method was employed to observe the patients. The most commonly observed activity was eating and drinking. The findings, in keeping with previous work, suggest that therapeutic activities occupy a minimal proportion of the patient's day. Reasons for inactivity are explored and recommendations made for further research. PMID- 8705083 TI - The supervision of Project 2000 students in the clinical setting: issues and implications for practitioners. AB - The aim of this 2-year study funded by the English National Board for Nursing, Midwifery and Health Visiting was to examine the relationship between teaching, support, supervision and role modelling in student clinical learning within the context of Project 2000 courses. The research, using a two-stage case-study design, employing predominantly qualitative methods of data collection, investigated the perceptions of students, tutors and practitioners of their experiences of these processes in the clinical setting. This paper focuses on the findings obtained from the practitioner data, high-lighting not only the significant role played by practitioners in facilitating student learning, but also the implications for practitioners in undertaking this role. The preparation required for supervising clinical learning, the integration of theory and practice and the organization of patient care have been identified as particularly important to the development of effective clinical learning environments. PMID- 8705084 TI - An investigation into the attitudes and practices of intensive care nurses towards verbal communication with unconscious patients. AB - This study attempted to investigate the attitudes and practices of intensive care nurses towards verbal communication with unconscious patients. A sample of five staff nurses working in an intensive care unit in Northern Ireland formed the basis for the study. The research design was non-experimental and descriptive exploratory in nature, incorporating 4-hourly observational periods and structured interviews. Qualitative and quantitative analysis indicated that intensive care nurses spend on average 5% of their time verbally communicating with unconscious patients. Most of this communication involves informing the patient of immediate procedural matters or providing reassuring statements. Most intensive care nurses claim that verbal communication with unconscious patients is very important, and some ambiguity is apparent as to the unconscious patient's level of awareness. Major factors influencing communication are the patient's level of consciousness, the amount of physical care being given and the presence of relatives. PMID- 8705085 TI - Creating consensus about nursing outcomes. I. An exploration of focus group methodology. AB - The aim of this study is to explore a method for achieving consensus about nursing outcomes between patients, nurses and health professionals. Focus group work as a research tool is described and reviewed. Problems of recruiting and facilitating such groups are highlighted. The relationship between focus groups and consensus is discussed. It is concluded that focus group work is not without problems, yet means of incorporating patient views into health services need to be developed. PMID- 8705086 TI - The wait, the wonder, the watch: caring in a transplant unit. PMID- 8705087 TI - Gender effect on diuretic response to hydrochlorothiazide and furosemide. AB - Gender has been shown to elicit differences in drug disposition and response to therapeutic agents. We measured the diuretic response to oral hydrochlorothiazide, oral and intravenous furosemide in 6 male and 6 female normal volunteers. After fasting overnight, each subject received single doses of the individual diuretics or no treatment on 4 separate days. Total urine output was collected over the next 24 hours for volume measurement and determination of sodium and potassium concentrations. There was no statistically significant difference found between male and female subjects with respect to urine flow rate, sodium, and potassium excretion rates among the treatments. However, when natriuretic response was adjusted for mg/kg of the intravenous furosemide dose received, male subjects had a higher peak sodium excretion rate than the female subjects. Results of this study reveal a gender-related difference on the natriuretic response to diuretics. Further studies are necessary to identify if this gender-related difference is caused by differences in drug metabolism, disposition, or intrinsic diuretic responsiveness at the site of action. PMID- 8705088 TI - Evaluation of the therapeutic range of whole blood cyclosporin concentration in the treatment of psoriasis. AB - Cyclosporin (CyA) trough levels in whole blood in 17 patients receiving CyA for the treatment of severe psoriasis were measured by fluorescence polarization immunoassay (FPIA) employing a specific monoclonal antibody. Clinical success was defined as a PASI score reduction greater than or equal to 75% (group A), partial success by a PASI score reduction of 60%-74% (group B), and insufficient efficacy by a PASI score reduction of less than 60% (group C). Nine of the 17 patients given CyA (53%) exhibited an improvement rate of > or = 75% in the PASI score within 8 weeks of the initiation of treatment. At 8 weeks of treatment the values for the daily dose and whole blood trough CyA concentration were both significantly higher in group A than in group C, daily dose (3.6 +/- 1.0 vs. 2.3 +/- 0.4 mg/kg/day, respectively) and whole blood concentration (141.3 +/- 46.0 vs. 79.7 +/- 19.0 ng/ml). Throughout the study period, during an average observation period of 30 weeks, 13 of the 17 patients (76%) achieved a reduction rate of > or = 75% in the PASI score. In these 13 patients in whom psoriasis lesions cleared satisfactorily the values for the mean daily CyA dose, and whole blood trough levels were 2.4 mg/kg/day and 94.8 ng/ml, respectively. The improvement rate was significantly higher when the CyA trough level was above, rather than below, 100-140 ng/ml within the first 8 weeks, although a lower CyA dose and/or whole blood level was effective in maintaining the reduced PASI score in the subsequent phase. The most important side-effect was mild hypertension in 7 of the 17 patients (41%), however, the onset of hypertension in these patients varied from 30-100 days after the initiation of treatment. Antihypertensive drug therapy and/or dose reduction resulted in a significant decrease, from 166 +/- 17/100 +/- 7-140 +/- 6/84 +/- 10 mmHg, of the CyA-induced hypertension. The relationship between the time course of the duration of improved PASI score and the whole blood concentration of CyA was observed, which suggests that the changes in drug effects were correlated with changes in whole blood levels. During the study period we experienced recurrences in 2 patients after the tapering or withdrawal of CyA treatment. In these 2 patients the relapse may have been due, in part, to the sharp decrease in whole blood concentration secondary to partial noncompliance. Measurement of whole blood CyA trough level is the only method for assessing the patient's noncompliance. Contrary to previous reports we confirmed that the measurement of CyA concentration in whole blood was useful for monitoring the effects in patients with psoriasis. PMID- 8705089 TI - Methods for monitoring and documenting adverse drug reactions. PMID- 8705090 TI - Chronopharmacokinetics of theophylline administered as a controlled-release tablet. AB - The result obtained from different studies of the chronopharmacokinetics of some controlled-release tablets of theophylline are variable, since some authors report differences while others do not. At our laboratory we have developed a formulation of a controlled-release theophylline tablet using acrylic resins and we studied the chronopharmacokinetics of theophylline from this dosage form. Seven Caucasian healthy male volunteers participated in the study approved by the Institutional Review Board (IRB). Each volunteer received a controlled release tablet of 300 mg theophylline and an i.v. dose equivalent to 131.46 mg theophylline once at 8.00 a.m. and once at 8.00 p.m. Theophylline plasma concentrations were determined by HPLC. The following pharmacokinetic parameters were calculated: maximum concentration, time to reach maximum concentration, mean residence time, absorption constant, area under the curve of plasma concentration versus time, distribution volume (Vd beta), and total clearance. No statistically significant differences were found between diurnal and nocturnal data. This implied that, with this formulation, there is a lower risk of toxic plasma concentrations or concentrations under the therapeutic level than with formulations that exhibit circadian rhythm. PMID- 8705091 TI - Effect of policosanol on hyperlipidemia and coronary heart disease in middle-aged patients. A 14-month pilot study. AB - To find out the long-term lipid-lowering efficacy of policosanol in low dose and its influence in the evolution of coronary heart disease (CHD), a pilot clinical randomized single-blind, placebo-controlled trial was conducted on 23 middle-aged outpatients, with well documented diagnosis of chronic CHD and primary or marginal hyperlipidemia. Twelve patients received policosanol tablets of 1 mg twice daily, and 11 patients placebo in the same fashion, followed with rest and stress electrocardiogram (ECG), and serum lipid blood samples by 14 months. The treated group showed significant reduction of total cholesterol in 14.8% (p < or = 0.001) and of low density lipoprotein (LDL) in 15.6% (p < or = 0.05), against non significant increase of 3% and 5.5%, respectively, in the placebo group. No patient had new coronary events in both groups, but 5 of 12 treated patients exhibited a clinical tendency to improve their CHD, in comparison with no one in the placebo group (p < or = 0.05). These findings show the effectiveness of low dose of policosanol lowering total cholesterol and LDL levels and suggest a CHD improvement in middle-aged patients with primary or marginal hyperlipidemia. PMID- 8705092 TI - Study of the bioequivalence of two controlled-release formulations of morphine. PMID- 8705093 TI - Assessment of cardiac performance: short- and medium-term variability of impedance cardiography at rest and during dynamic exercise. AB - Short- and medium-term variability of impedance cardiography at rest and during exercise at a heart rate of 155-160 beats/min were assessed in 12 healthy men aged 21-28 years. Two consecutive measurements within 1 min were performed 4 times at 2-hour intervals on 2 days 14 days apart. Ejection fraction was the most reproducible of all impedance cardiography parameters, the CV ranging from 3.3 5.9%. The short-term reproducibility of cardiac output and stroke volume at rest was good, the between-repeats coefficient of variation (CV) being 4-6%. The reproducibility weakened with longer time span (between-day CV being about 12%) and higher heart rates (exercise). However, even the between-day reproducibility at rest was as good as that of heart rate. Between-day CV during exercise were about 20%. PMID- 8705094 TI - The effect of a 1-week administration of cetirizine on the chemotaxis and superoxide anion production of neutrophils from healthy volunteers. AB - The aim of the present investigation was to examine the effect of a 1-week oral administration of the anti-allergic drug cetirizine on healthy volunteer neutrophil chemotaxis and superoxide anion (O2-) production. Eight male volunteers were selected after clinical examination and laboratory tests. Neutrophils were isolated from peripheral blood using a discontinuous density gradient. Spontaneous migration and platelet activating factor (PAF, 10(-6) and 10(-8) M) or zymosan-activated plasma (ZAP)-induced chemotaxis were studied in a 48-well microchemotaxis chamber. Basal and phorbol 12-myristate 13-acetate (PMA, 30 nM) stimulated O2- production were measured spectro-photometrically. Cetirizine (10 mg per day) was given orally during 1 week. Both neutrophil chemotaxis and O2- production were assessed before and 2 h, 24 h, and 1 week after orally administered cetirizine. Plasma cetirizine levels were monitored by high performance liquid chromatography (HPLC) with ultraviolet detection. Spontaneous neutrophil migration and PAF-or ZAP-induced chemotaxis showed no significant variation before or at various intervals after the initiation of treatment with cetirizine. Basal and PMA-stimulated neutrophil O2-production was also not affected by cetirizine. The maximum concentration attained by cetirizine (Cmax) was 293 +/- 38 ng/ml and generally peaked (Tmax) within 1.9 +/- 0.7 h. We conclude that the administration of cetirizine for 1 week does not alter human neutrophil chemotaxis and O2- production. PMID- 8705095 TI - Endoscopic carpal tunnel release using a single-portal system. AB - In this prospective clinical study, we present our experience with a new single portal carpal tunnel release kit. The safety and efficacy of this new device was assessed and compared with a consecutive control group treated with conventional open carpal tunnel release. 126 patients were enrolled in this study, 64 of them were treated endoscopically (group 1) and 62 by open release of the carpal ligament (group 2). Follow-ups were conducted at 1, 3, 6, 12 and 24 weeks postoperatively. A serious intraoperative complication in group 1 was a transection injury of a branch of the superficial palmar arch. No intraoperative complications were noted in group 2. Postoperative evaluation revealed significantly less scar tenderness in group 1 at 1 week (P < 0.001), 3 weeks (P < 0.001) and 24 weeks (P < 0.05) compared to group 2. Functional status at 1 week was significantly (P < 0.05) better in group 1 than in group 2 but not at later times. Grip strength at 1 week (P < 0.001), 3 weeks (P < 0.05) and 12 weeks (P < 0.05), and pinch strength at 3 weeks (P < 0.001) were significantly higher in group 1. No significant differences between the groups were obtained regarding postoperative symptom severity. The new device provides a reliable tool for single portal carpal tunnel release, although the risk of inadvertent damage to the neurovascular structures always remains a possibility with the endoscopic carpal tunnel technique. PMID- 8705096 TI - Measurement of sudomotor fibre regeneration by sympathetic skin response after complete division of peripheral nerves in children. AB - Sympathetic Skin Response (SSR) was measured in 14 paediatric patients (age range 3-15 years, mean 7.2 years) following repair of lacerated upper limb peripheral nerves (median, ulnar or both). All nerves had been completely divided and were repaired by primary epineural repair. Measurements of SSR were made at regular, 1 2 monthly, intervals during nerve regeneration in 6 patients. A further group of 7 patients had single SSR measurements at the end of nerve regeneration (21-63 months). A 4-year-old boy was also followed up from 10 to 21 months following replantation of a proximally amputated right upper limb. The method was first standardised in 16 healthy volunteers (age range 3-17 years, mean 9.4 years). Patients over 6 years of age were also clinically tested for return of sensation. The results show that the objective measurement of sudomotor nerve regeneration in children is possible with this method. They demonstrate its universal acceptance by children as young as 3 years old. PMID- 8705097 TI - Asymmetrical cleft palate. AB - Ten cases of asymmetrical cleft palate are reported. The pathogenesis, classification and management are discussed. PMID- 8705098 TI - Breast feeding or spoon feeding after cleft lip repair: a prospective, randomised study. AB - It has been a tradition for plastic surgeons to withhold breast feeding from babies after cleft lip repair to prevent wound disruption. A prospective, randomised trial of 40 infants showed that early postoperative breast feeding after cleft lip repair is safe, results in more weight gain at 6 weeks after surgery (P < 0.01) and is more economical than spoon feeding. PMID- 8705099 TI - Soft tissue sarcomas: diagnosis, classification and prognostic factors. PMID- 8705100 TI - A morphological study of the long-term repair process in experimentally stretched but unruptured arteries and veins. AB - As many avulsion amputations are incomplete and the vessels remain intact, the immediate pathology and long-term repair process (to 3 months post-injury) of experimentally stretched but unruptured rabbit femoral arteries and veins were examined. In stretched arteries, circumferential skip lesions involving endothelium, internal elastic lamina (IEL) and media occurred frequently and often up to 3 cm from the point of stretch. Medial smooth muscle cells (SMC) were significantly damaged or lost at lesions. Macrophages and neutrophils were found in lesions 1-4 days post-injury. Between 2-4 days, lesions were covered by endothelium and synthetic state SMC appeared in the media. At 1 week, a thin neointima (which persisted to 3 months) covered many lesions. The media at lesions gradually filled with SMC but generally remained disorganised even at 3 months post-injury. Stretching caused tears in vein walls, particularly close to the point of injury. There was no evidence of venous damage or repair in specimens examined 3 weeks and 3 months post-injury. PMID- 8705101 TI - The microcirculation and survival of experimental flow-through venous flaps. AB - The subject of venous flaps is controversial. The purpose of this study was to investigate the haemodynamic changes and the survival process of venous flow through flaps. Venous flaps and composite grafts were elevated along the axis of the thoracoepigastric veins in rabbits. The microcirculation was studied by capillaroscopy and injection studies. For the first 72 h, the flaps were nourished by venous blood; venous blood flowed slowly from the principal vein to its tributary veins and then through venous anastomoses to reach other tributary veins. No backflow from venules into capillaries was seen. New vessels began to grow into the flaps from 72 h postoperatively. Four days postoperatively, arterial flow with low velocity was seen in the primary flap arteries and the backflow of venous blood disappeared. Seven days postoperatively, the abnormal flow in "unphysiological" channels was replaced by normal flow in "physiological" channels. Six weeks postoperatively, the vessel structure of venous flaps was the same as that of normal skin. These results suggest that the profuse venous network in venous flaps and early invasion of new blood vessels are the mainstays of venous flap survival. PMID- 8705102 TI - A modified surgical technique for temporalis transfer. AB - A modification of the temporalis muscle transfer for facial animation in facial palsy is described. Seven cases are presented which demonstrate the advantages of the procedure, especially the avoidance of temporal hollowing and the greater power due to less dissection of the muscle belly, a more direct line of pull and the absence of a pulley. In addition, food pooling in the buccal sulcus is reduced, and the procedure is less extensive and thus better tolerated. PMID- 8705103 TI - The lateral calcaneal artery adipofascial flap. AB - The lateral calcaneal artery adipofascial flap is a modification of the lateral calcaneal artery skin flap. It has been used successfully in five patients. In comparison to the skin flap, it has the advantages of more versatile clinical applications, better aesthetic result of the donor site and sural nerve preservation. PMID- 8705105 TI - Fascial release as an adjunct to wound closure. AB - We describe a simple technique to facilitate the primary closure of skin defects. Skin, subcutaneous tissue and fascia are undermined as a unit on both sides of the defect. Fascial releasing incisions are performed, creating two bipedicled fascial flaps attached to the overlying skin. The fascia is approximated, if possible, and the skin sutured. We believe this technique offers a number of advantages and results in a superior closure to that obtained with the conventional method of skin undermining. This approach has enabled the closure of 10 of 11 medium sized wounds. PMID- 8705104 TI - Acceleration of granulation tissue ingrowth by hyaluronic acid in artificial skin. AB - Hyaluronic acid (HA), which is known to play an important role in wound healing, was incorporated in an artificial skin material and studied for its potential to create a wound bed which would support a skin graft. Collagen sponge based artificial skin was soaked in 0.3% HA in phosphate buffered saline and grafted onto skin defects in rats. Control grafts were soaked in normal saline solution. HA incorporated implants and control implants were simultaneously grafted onto wounds made on either side of the spine. To examine the effect of HA incorporation, the percentage area of cellular tuft infiltration and the number of capillaries present in the graft matrix were evaluated at 7 and 14 days after the operation. At postoperative day 7, there was a statistically significant difference in the number of capillaries in the matrix of the experimental versus the control implants. There was no difference in the percentage area of cellular tuft infiltration. At postoperative day 14, all implants exhibited better ingrowth of granulation tissue than at day 7. The differences between the experimental and control implants were statistically significant with respect to both the percentage area of cellular tuft infiltration and the number of capillaries. It is therefore concluded that in artificial skin HA incorporation accelerates the ingrowth of granulation tissue, making a more suitable graft bed. PMID- 8705106 TI - Metastatic colonic adenocarcinoma in a pedicled omental flap used for sternal reconstruction: a case report. AB - Dehiscence of a median sternotomy wound is a fortunately rare but potentially lethal complication of cardiac surgery. If conservative management, including irrigation and secondary closure, fails then osteomyelitis with or without necrosis of the sternum, costochondritis and anterior mediastinitis may result. In the face of such sequelae, radical debidement of the sternum and flap coverage is required. A variety of flaps may be used to cover the defect following debridement of the sternum, in particular pectoralis major or rectus abdominis muscle flaps and the omentum. We report the case of a 65-year-old man, who required an omental flap for sternal dehiscence after coronary artery bypass grafting and who subsequently presented with a metastatic colonic adenocarcinoma within the flap. PMID- 8705107 TI - Postauricular chondrocutaneous flap in auricular reconstruction. PMID- 8705108 TI - Recent clinical experience with cultured autologous epithelium. PMID- 8705109 TI - Cleft lip and amniotic bands. PMID- 8705110 TI - Facelifting: an initial eight year experience. PMID- 8705111 TI - Standard operating procedures (SOPs): reason for, types of, adequacy, approval, and deviations from and revisions to. AB - Standard Operating Procedures (SOPs) are required in order to comply with the Good Laboratory Practice Standards (GLPS) as promulgated in the Federal Insecticide, Fungicide and Rodenticide Act (FIFRA) 40 CFR Part 160. Paragraph 160.81 (a) states: "A testing facility shall have standard operating procedures in writing setting forth study methods that management is satisfied are adequate to insure the quality and integrity of the data generated in the course of a study." Types of SOPs include administrative and personnel, analyses, substances, quality assurance and records, test system, equipment, and field related. All SOPs must be adequate in scope to describe the function in sufficient detail such that the study data are reproducible. All SOPs must be approved by a management level as described in a corporate organization chart. Signatures for SOP responsibility, authorship, and Quality Assurance review adds strength and accountability to the SOP. In the event a procedure or method is performed differently from what is stated in the SOP, an SOP deviation is necessary. As methods and procedures are improved, SOP revisions are necessary to maintain SOP adequacy and applicability. The replaced SOP is put into a historical SOP file and all copies of the replaced SOPs are destroyed. PMID- 8705112 TI - The changing role of quality assurance: an international industry perspective. AB - As a Scientific Consultant in the area of Good Laboratory Practice and Good Clinical Practice the author has been involved with Quality Assurance (QA) for 18 years and fells the time has come for Industry to explore a better way to utilize the many talents that the QA professional has achieved. They have a wealth of expertise and should become more involved in the integration of "science" with the many "GXPs" to ensure that compliance is arrived at on a cost benefit basis for the company. PMID- 8705113 TI - Quo vademus? A review of the regulatory environment. PMID- 8705114 TI - Quantitative structure-activity relationships for estimating the no-observable effects concentration in fathead minnows (Pimephales promelas). AB - Quantitative structure-activity relationships, based on hydrophobicity measured as the 1-octanol/water partition coefficient (Kow), have been developed for predicting the acute no-observable-effects concentration (NOEC) for fathead minnows (Pimephales promelas), using a mechanism/mode of action approach. Models were developed for nonpolar narcotics, amine narcotics, polar narcotics, ester narcotics, and respiratory uncouplers. A set of bioreactive chemicals were evaluated, but models based on hydrophobicity alone are inadequate for these chemicals. Additionally, a strong correlation was observed between the acute NOEC and acute lethality measured as the 50% lethal concentration (LC50) for 407 chemicals, regardless of the mechanism/mode of action. This relationship is modeled by the equation log NOEC-1 = 1.007 (log LC50) + 0.246; n = 407, r2 = 0.989, s = 0.142, f = 38095.6, and Pr > f = 0.0001. Examination of the relationship between the acute NOEC and the subchronic NOEC for four mechanisms/modes of action implies that the acute to subchronic ratio is not independent of mechanism of action. The resulting ratios were 6.42 (+/- 3.01) for nonpolar narcosis, 80.00 (+/- 17.78) for polar narcosis, 10.05 (+/- 4.59) for respiratory uncoupling, and 3.94 (+/- 1.72) for bioreactivity. PMID- 8705115 TI - Quality initiatives in a corporate toxicology laboratory. AB - Quality improvement initiatives for a corporate toxicology group require a high level of managerial commitment to a bring about lasting change, a well-enunciated plan, professional training, effective communication, and reward and accountability systems. Learnings are shared which should be applicable to other toxicology or corporate staff groups. PMID- 8705116 TI - Best practice in workplace hazardous substances management. AB - Chemical-induced injury and disease remains a significant problem in workers in industry. As a result of this problem, a number of national and international initiatives have recommended the development of conventions, regulations, and codes of practice to attempt to deal with the problems of hazardous substances at work. Within Australia, workplace hazardous substances regulations are in development which will impose legal obligations and responsibilities on the suppliers of hazardous substances and on the employers who use them. At the same time, internationally consistent ISO standards are in use, or are being developed, for quality systems, environmental management, and occupational health and safety. These standards outline a model for the management of quality, environment, or safety, and the processes involved are applicable to the management of hazardous substances. This process includes: obtaining commitment from senior management; instituting consultative mechanisms; developing a hazardous substances policy; identifying components of the hazardous substances management program; resourcing, implementing, and reviewing the program; and integrating the program into the organisation's strategic plan. Only by blending in a specific management program for hazardous substances into the overall planning of an organization will they be managed effectively and efficiently. PMID- 8705117 TI - FDA audit of nonclinical studies. AB - FDA is increasing its efforts to enforce regulations governing the conduct of nonclinical (animal) trials to support evidence of safety. Since the GLP regulations were promulgated in 1987, the incidence of deficiencies at these animal facilities has remained constant despite an increased number of issued warning letters. These warning letters constitute a serious breach of the FDA regulations and has resulted in certain cases in rejection of nonclinical study data, termination of the IND application, and disqualification of the facility as well as criminal prosecution of individuals involved in fraudulent practices. PMID- 8705118 TI - [Seroprevalence of toxoplasmosis in Dakar (Senegal) in 1993: study of women in their reproductive years]. AB - To establish the seroprevalence of toxoplasmosis in women during their reproductive years, we examined 720 women, of whom 404 were pregnant and 306 were not, all residing in and around Dakar. Of the serum tested by indirect immunofluorescence, 40.3% contained antibodies recognizing toxoplasmosis. The seroprevalence did not vary significantly with the age of the women, the number of previous pregnancies, the place of residence or the length of residence in Dakar. It was higher in the pregnant women (44.4%) than in the nonpregnant women (37.2%), but this difference is not statistically different. The titers of antibodies were generally weak, varying between 10 and 320 IU/ml of serum. Also, IgM specific antibodies were absent. These results indicate that the seroconversion had occurred previously, probably at a young age. We observed seroprevalences higher than those obtained by authors using comparable techniques twenty years ago. This indicates an increase of the transmission of toxoplasmosis in the population. Few women during their reproductive years had antibodies against toxoplasmosis. Thus, they are at risk of developing a primary infection during a pregnancy. This undergoes the necessity of promoting measures to prevent toxoplasmosis infection in pregnant women. PMID- 8705119 TI - [Quality of health services in Africa : example of dystocia prenatal screening in Nioki (Zaire)]. AB - Childbirth, though a common yet highly valued social event in Africa, is a major risk for the mother and child. During these last few decades, extension of health services has gained attention and their accessibility has improved. However, these health services remain inefficient because of their poor quality, especially for prenatal screening. This study with 59 cases and 177 controls was carried out in a rural district of Zaire. The objectives were to first assess the quality of the diagnosis of high risk pregnancy (HRP) and secondly to identify the main risk factors associated with complicated deliveries. The diagnosis of HRP has a low sensitivity (53%) yet a high specificity (95%). The diagnosis is based on postpartum hemorrhage and maternal height but does not consider primiparity or previous complicated deliveries. By improving the use of currently available information, a higher sensitivity (72%, could be obtained, identifying close to 3 out of 4 HRPs, but with a lower specificity (67%). The current diagnosis is conservative, partly because health personnel are reluctant to refer pregnant women, who could deliver normally, to the district hospital. Predictive criteria of complicated pregnancy are: primiparity, maternal height, previous difficult childbirth (dystocia), previous postpartum hemorrhage and lack of prenatal care. Prenatal screening should be a major component of safe motherhood programs. Our results show that in developing countries where emphasis is mainly put on the accessibility of health services, the quality of these services should become a priority. Quality is linked to efficiency, yet good quality gives a better image of services and improves compliance of the women. PMID- 8705120 TI - [Development and evaluation of screening algorithms for sexually transmitted diseases in pregnant women at Libreville, Gabon]. AB - The struggle against sexually transmitted diseases (STD) constitutes a priority of public health in developing countries: STD cause complications, particularly in pregnant women, and facilitate the transmission of HIV. One of the strategies in the struggle against STD is the diagnosis and the early treatment of these infections. The STD, and in particular infections of Neisseria gonorrhea and Chlamydia trachomatis, are difficult to diagnose in women without complementary analyses, which primary health care may not be able to supply. Health care provided to patients could be standardized and improved by considering the signs and symptoms. We studied the prevalence and risk factors of STD among 192 pregnant women consulting the health clinic in Libreville, Gabon, in September 1993. The prevalence of STD was high (13.5% rate of cervical infection with gonorrhea or Chlamydia trachomatis). We then evaluated the different diagnostic strategies or algorithms. Regardless of the type of examination (medical interview, simple clinical examination or examination with a speculum), the use of scores integrating risk factors, the clinical signs and symptoms outperformed hierarchical algorithms. This approach was more sensitive and specific and easy to perform. Use of this method may enable more effective screening of STD and also avoid most maternal and perinatal complications. PMID- 8705121 TI - [Foreign international bibliographic databases: Index Medicus. Presentation and user instructions]. AB - Index Medicus, edited by the National Library of Medicine, Bethesda, MD, USA, is a bibliography of the international biomedical literature and indexes more than 3,000 scientific reviews. The printed edition contains five sections: a list of indexed journals, the medical subject headings, a bibliography of medical reviews, a subject section and an author section. We explain how to conduct bibliographical searches within these five sections. The list of indexed journal shows the reader the abbreviated title in addition to the full title for writing a citation, as well as most of the important journals indexed by subject and the editing countries. Authors names are simply retrieved within the author section. Subjects are located first under the medical subject headings which contain the keywords, and then under the bibliography of medical reviews or the subject section. Despite the emergence of computer-aided literature searches, the printed data base of the Index Medicus remains a useful tool in the medical libraries around the world. PMID- 8705122 TI - [AIDS and drug addiction]. PMID- 8705123 TI - [Methadone in France. The experience of the Monte Cristo center of the Laennec Hospital in Paris]. PMID- 8705124 TI - [Strategy for risk reduction of HIV infection in drug addicts in South Vietnam]. PMID- 8705125 TI - [HIV epidemiology among drug addicts in the city of Ho Chi Minh. Trends between 1992 and 1995]. PMID- 8705126 TI - [Clinical dermatologic and genital manifestations in drug addicts infected with HIV]. PMID- 8705127 TI - [Epidemiologic trends of HIV infection in South Vietnam]. PMID- 8705128 TI - [Clinical and epidemiologic aspects of HIV infection among drug addicts in Ho Chi Minh city]. PMID- 8705129 TI - [AIDS epidemiology and intravenous drug use in Europe]. PMID- 8705130 TI - [Tuberculosis in patients infected with HIV in Ho Chi Minh city]. PMID- 8705131 TI - [Somatic pathology in drug addicts infected with HIV]. PMID- 8705132 TI - [Reduction in crimes of drug addiction in North America]. PMID- 8705133 TI - [AIDS and drug addiction in North America. Drug addiction epidemiology]. PMID- 8705134 TI - [Rice: source of life and death on the plateaux of Madagascar]. AB - Since the 17th century, Europeans travelling in Madagascar described the contrast between the fever-free Plateau and the fever-ridden coasts. The former were inhabited by people of Asiatic origins and the latter by African migrants. At the end of the 18th century, "Merina" kings developed land irrigation and rice cultivation, using manpower from the coasts. Since then, rice has become a monoculture covering most of the arable lands of the Highlands. The first malaria epidemic occurred in the Tananarive area in 1878, and rapidly spread throughout the Plateau. The mortality rate was high. A second epidemic in 1895 may have been a resurgence of the previous one. Subsequently, malaria became meso-epidemic despite control measures, mainly consisting of larvivorous fishes, quinine treatment and prophylaxis. In 1949, an eradication program was launched based on DDT house-spraying and chloroquine prophylaxis in children. It was very successful on the Highlands where malaria disappeared, in 1962. Spraying was cancelled and only three small foci remained under surveillance. In 1987 and 1988, a malaria outbreak devastated the plateau. Subsequently, intensive spraying operations brought the situation under control by 1993. The main malaria vector on the Madagascar Highlands is An. funestus. More than 95% of its breeding sites are in the rice fields just before the harvest and afterwards in the fallow lands. The vector peak and the corresponding peak of malaria cases occur between February and May, depending on the farming calender. The second but less important vector, An. arabiensis, breeds in the rice fields just after seeding when the surface water is sunlit. Although rice fields remain the main source of this vector, it also breeds in rainwater pods and borow-pits. Malaria vectors on the plateau are products of human activities of rice cultivation, which is the basis of the economy. The epidemiological importance of rice fields varies greatly from one country to another. In Southeast Asia, the rice fields harbor several anopheline species most of which are only vectors of P. vivax. In West Africa where malaria is holoendemic, they produce large populations of An. gambiae; however, the malaria pattern is unaltered and remains at peak levels. In the dry areas of southern Madagascar, the vector An. funestus and meso hyperendemic malaria are restricted to areas of cultivated rice. In West and Central Africa, An. funestus is never found in rice fields even though it is common in marshes. In Madagascar, this vector breeds in irrigated rice fields. Because it is practically impossible to control anophelines in rice fields by chemical, biological and ecological methods on the Highlands of Madagascar, house spraying remains the best method for mass malaria control. Bed-nets impregnated with pesticides may offer an alternative, but their use is resisted by the local population. PMID- 8705135 TI - [Control of an epidemic of meningococcal meningitis in Central Africa]. AB - A severe epidemic of group A meningococcal disease occurred in the northwest part of the Central African Republic from January to March 1992. The outbreak affected a large and densely populated area, with a poor road network, located 400 kilometers south of the classical meningitis belt. An initial selective vaccination campaign was carried out by the national health care service. As the epidemic was continuing, the national authorities asked for international assistance. The French participated by sending Bioforce, a medical task force designed by the Ministry of Defense, with the financial support of the Ministry of Cooperation. Neisseria meningitidis strains were isolated and identified within 36 hours by the Bioforce field laboratory. Strains from 24 patients were sent to the Pasteur Institute in Paris (Neisseria Unit) for serotyping, testing of antibiotic susceptibility, and multilocus enzyme electrophoresis. With one exception, all strains had formula A:4:P1.9. By an initial rapid assessment, the limits of the affected area and populations were determined. The weekly incidence rates observed in different areas varied within a range of 3 to 10 cases per 1,000, with fatality rates from 20 to 30 cases per 100. The spread of the epidemic was stopped by a mass vaccination campaign, which targeted the entire population (200,000 immunizations) of the affected area. The case fatality rate could not be reduced below 15%, despite antimicrobial treatments implemented as soon as possible. The optimal treatment was the standard single intramuscular injection of oily chloramphenicol. The predictive values of clinical symptoms were calculated. The efficacy of vaccination was estimated by comparison of the percentage of people immunized and the proportion of those vaccinated people who developed meningitis identified during the 3 weeks following the mass vaccination campaign. The efficacy varied between 93 to 95% according to the place. The typical weekly incidence rate of 1 case per 1,000 is not a relevant threshold to sufficiently and immediately detect a meningitis outbreak and needs to be reconsidered. PMID- 8705136 TI - [Evaluation of malaria vector control measures in central Vietnam (1976-1991)]. AB - Activities used to control malaria transmission in the pilot station of Vanh Canh in the Binh Dinh Province of central Vietnam from 1976 to 1991 have been evaluated. These activities were: spraying DDT in and around the houses in the villages and the settlements in the fields; spraying lambdacyalothrin in the houses; and use of bed-nets impregnated with permethrin. Their efficacy was measured by the number of fever episodes due to malaria infections among the population. The spraying of DDT in the houses was followed by a reduction of malaria infection by more than 90%. However, spraying of the settlements was not advantageous. The termination of DDT spraying was not followed by an increase of malaria infections. Spraying with lambdacyalothrin was slightly more effective than with pyrimiphos and DDT. The use of pesticide-impregnated bed-nets was efficient, especially in the villages far away from the forest. Thus, these activities can contribute to the control of the malaria endemic in central Vietnam. PMID- 8705137 TI - [Hygienic prophylaxis: problems and solutions]. PMID- 8705138 TI - [Mortality of geologists in relation to occupational activities]. AB - The study covered mortality among a cohort of geologists working in North-Western Russia. The cohort included 3911 geologists, geophysicists and drillers, whose life was followed during 1977-1992. The standardized relative mortality risk with all death causes equalled 0.58 in general. Risk of mortality with hypertension in the male geologists was 21.7 times higher and for the female ones-25.4 times higher than for general population. Definite dependence between the stronger effects and the longer length of service was seen. The highest standardized relative mortality risk appeared among the drillers. Occupational risk factors for geologists reliably increase risk of mortality with active rheumatism, diseases involving arteries, veins and lymphatic vessels. The male geophysicists tend to have higher incidence of prostatic cancer, and female ones-higher incidence of uterine carcinoma. PMID- 8705139 TI - [Study of the role of parents' occupation in children's mortality]. AB - This study have been carried out to: a) investigate the prevalence of such occupational factors in parents which could be fatal to their children, and b) evaluate quantitatively the risk of mortality in infants caused by some leading diseases developed as a result of harmful occupational factors to which their parents had been exposed. During this work the data bank on infant mortality in Tallinn based on deaths registration in city archive for period 1968-1992 has been created. This bank includes the data on parents' occupation. The prevalence of parents' occupational risk factors has been analyzed in infants who die due extreme immaturity and other prematurity cases, congenital anomalies neoplasias and control group persons. To evaluate the death risk in children the case control epidemiological study with the individual group selection has been performed. This pilot analytical epidemiological study allows to suppose involving of possibly harmful occupational factors which are playing a certain role influencing mortality in whole contingent investigated, but not concerning individual nosologic forms. For the final judgement concerning the role of occupational risk factors the additional special fundamental study including the extended number of observations and the data, on interrupting nonoccupational risk factors prevalence should be carried out. PMID- 8705140 TI - [Toxic pneumosclerosis and alveolitis in workers of hydrometallurgic production of nickel]. AB - The author detected a considerable occurrence (14.4%) of toxic pneumosclerosis resulting from extrinsic toxic alveolitis in workers engaged into hydrometallurgic nickel production. Pathogenesis of the condition includes changes in lung surfactant, dysbalance between lipid peroxidation and antioxidant activity, immune disorders, formation of circulating immune complexes. PMID- 8705141 TI - [Improved work conditions in present-day porcelain production]. AB - The authors studied work conditions at Doulev porcelain production after its reconstruction. The dust levels appeared to decrease, therefore possible occurrence of occupational diseases caused by dust became lower. A problem of manual tasks liquidation still remains topical. PMID- 8705142 TI - [Element levels of blood in workers of the Matrosov mine]. AB - The article deals with results obtained during the study of serum microelements among workers of the mine and gold-extracting factory in Central Kolyma. Atomic emission spectroscopy evaluated over 20 elements, the maximal information was obtained on Ca, Mg, Fe, Cu, Zn, Cd, Se, Mn, Pb, Hg. The examinees were diagnosed as having lower levels of Ca, Mg, Se, Zn and increased ones of Cu, Pb, Hg, Cd, so iron and zinc deficit could occur. The studies bring to a conclusion that the workers suffer from complex polymicroelementosis, the more intensive the longer life in the North. Additional examination is required for the individuals having increased serum Hg, Cd and Pb levels. PMID- 8705143 TI - [Percutaneous laser irradiation of blood in the treatment of hemodynamic disorders in patients with vibration disease]. AB - The article presents data on influence of percutaneous laser irradiation of blood on central and peripheral hemodynamics in 24 riveters facing vibration disease. Clinical improvement was seen in all the examinees. Rheography showed high efficiency of the method in the patients with initially increased vascular tone in hands (angiospastic syndrome), possible correction of disordered cerebral circulation. PMID- 8705144 TI - [Physiologic and health evaluation of work capacity of pilots during daily shifts]. PMID- 8705145 TI - [Early diagnosis of metabolic disorders in the gastric mucosa of flight personnel]. PMID- 8705146 TI - [Cytogenetic effects of zeolites extracted from various grounds]. PMID- 8705147 TI - [Information on the activities of the special Education-Methodology Committee on Occupational Diseases]. PMID- 8705148 TI - [Role of visible irradiation in industrial medicine]. AB - Modern norms regulating illumination levels of working surfaces do not consider changes in functional state of vision. Experimental studies involving examinees helped to set optimal levels of brightness for light entering eyes in complicated visual work. If larger object viewed, brightness of the working surface could be lowered to values that do not decrease visual efficiency. The data obtained enabled to justify allowable brightness levels in accordance with complexity of visual tasks. Regulation of visible light according to brightness values appears to be more adequate for visual functions. PMID- 8705150 TI - Ventilatory responses to hypoxia and hypercapnia in relation to anesthetic practice. PMID- 8705149 TI - [Productivity of modelled work of near-sighted microscope operators under variable lighting]. AB - A total of 10 examinees with shortsight at 1.0-3.5 diopters were subjected to study of visual performance similar to work at microscope with regular and chromatic (590 nm) light to tested objects. Chromatic light appeared to result in reliably higher visual performance, especially by the end ot 40-minute experiment. The study used an original method modelling visual work and a formula to calculate performance of work at microscope. PMID- 8705151 TI - Comparison of antihypertensive effects of nicardipine with nitroglycerin for perioperative hypertension. AB - BACKGROUND: To compare the efficacy of intravenous (iv) nicardipine with nitroglycerin for the treatment for patients with perioperative hypertension. METHODS: Forty patients with perioperative hypertension randomly divided into two groups were treated with intravenous calcium entry blocker, nicardipine, or vasodilator, nitroglycerin. Haemodynamic measurements including mean arterial and pulmonary arterial pressure, central venous and pulmonary capillary wedge pressure, and cardiac output were recorded; peripheral and pulmonary vascular resistance were calculated. RESULTS: Both medications were effective in reducing blood pressure and controlling haemodynamics. During the maintenance by continuous iv infusion, nicardipine controlled hypertension more rapidly than nitroglycerin (nicardipine 10.5 +/- 2.5 min and nitroglycerin 18.7 +/- 2.8 min, p < 0.05) without significant alteration in heart rate. The total frequency of dose adjustments required to achieve therapeutic response was significantly less in the nicardipine-treated group (2.5 +/- 0.3 for nicardipine and 6.2 +/- 1.4 for nitroglycerin, p < 0.05). Incidence of hypotensive episodes during the infusion were observed in both groups [nicardipine 5% (1/20) and nitroglycerin 30% (6/20), p < 0.05]. CONCLUSIONS: Intravenous nicardipine is as effective as nitroglycerin in the treatment of perioperative hypertension. Specific advantages have been identified such as stable dose-response effect, less hypotensive and tachycardial effects during the use of iv nicardipine in treatment of hypertensive patients. PMID- 8705152 TI - The use of intranasal nitroglycerin to prevent pressor responses during intubation in general anesthesia--a comparison of various doses. AB - BACKGROUND: Intranasal nitroglycerin (NTG) was first reported to successfully prevent an increase in arterial blood pressure following laryngoscopy and tracheal intubation by Hill et al. Various different effective dosages of NTG have been reported. Grover et al. indicated 0.75 mg of intranasal NTG to be the most suitable dose. However, no definite conclusion has yet been made. This study was designed to compare the efficacy of four different dosages of intranasal NTG (0.3, 0.5, 0.75, and 1.0 mg) in preventing pressor responses to laryngoscopy and tracheal intubation during the induction of general anesthesia. METHODS: One hundred patients (ASA I or II) scheduled for elective surgery were included. These study subjects were divided into five groups and randomly assigned to four different dosages of intranasal NTG and a placebo. Each group consisted of 20 patients. The NTG solution was administered 1 min before the injection of thiopental. Systolic arterial pressure (SAP), diastolic arterial pressure (DAP), mean arterial pressure (MAP) and heart rate (HR) were recorded before the induction of anesthesia (T1), before laryngoscopy (T2), and at 0, 3, and 5 min after tracheal intubation (T3, T4, and T5 respectively). RESULTS: In patients who received a placebo (control group), there were significant increases in SAP, MAP, HR and rate-pressure-product (RPP) associated with tracheal intubation. Tachycardia was noted in all experimental groups. The increases in MAP associated with tracheal intubation were significantly less in patients who received NTG of 0.5 mg or more but not 0.3 mg. Although 0.5 mg of NTG did attenuate the increases in SAP after tracheal intubation, the increases in SAP of the other three experimental groups were no less than that of the control group. Rate-pressure product (RPP) values of the experimental groups were noted to be equal to or higher than those of the control group during the period of study. Contrary to the results of the study conducted by Grover et al., 0.75 mg of NTG did not attenuate the pressor responses. CONCLUSIONS: Intranasal NTG does not attenuate the pressor responses to laryngoscopy and tracheal intubation. PMID- 8705153 TI - The preemptive effect of regional anesthesia on post-cesarean section pain. AB - BACKGROUND: The treatment of post-cesarean section (C/S) pain is very important. Since no previous report discussed the preemptive effect of regional anesthesia on post-C/S pain, the present study was undertaken to evaluate the efficacy of the preemptive effect of regional anesthesia on post-C/S pain. METHODS: Ninety parturients scheduled for C/S were enrolled into one of the three groups (n = 30, per each group). The parturients in group 1 and 2 received spinal anesthesia and epidural anesthesia, respectively, while those in group 3 received general anesthesia for C/S. Postoperative patient-controlled analgesia, by means of intravenous morphine, was used for all paturients to relieve post-C/S pain. The VAS of pain and total morphine consumption were evaluated in all 3 groups of patients. RESULTS: The results showed that groups 1 and 2 consumed less morphine than that of group 3. Also, there was no difference between groups 1 and 2 in the amount of morphine consumed. The incidence of side effects was significantly higher in group 3 than in groups 1 and 2. CONCLUSIONS: We demonstrated that spinal or epidural anesthesia reduced the need of parenteral morphine postoperatively when compared with general anesthesia. We suggest that the use of regional anesthesia may provide a better preemptive effect for post-C/S pain. PMID- 8705155 TI - Ventilatory responses to hypoxia and hypercapnia during inhalation anaesthesia and subparalytical muscle relaxation in humans. PMID- 8705154 TI - Opioid in peripheral analgesia: intra-articular morphine for pain control after arthroscopic knee surgery. AB - BACKGROUND: Some authors reported that a small dose of intra-articular morphine (1-5 mg) injection provided a potent and long-acting analgesic effect on pain after arthroscopic knee surgery. However, many in other reports did not agree to this result. Therefore, the characteristic of the analgesic effect of intra articular morphine is worth evaluation. In order to rule out the systemic action of intra-articular morphine, we designed a study to compare the efficacy of 3 mg intra-articular morphine with that of 3 mg intravenous morphine in providing analgesia after arthroscopic knee surgery. METHODS: Sixty patients undergoing arthroscopic knee surgery under spinal anesthesia were randomly divided into two groups (n = 30, each). At the end of surgery, group 1 received 3 mg intra articular morphine in 15 ml isotonic saline and intravenous isotonic saline 10 ml; group 2 received 15 ml intra-articular isotonic saline and 3 mg intravenous morphine in 10 ml isotonic saline. Postoperative pain was evaluated using visual analogue scale (VAS) during an active or passive flexion of the operated knee joint. Also, additional analgesic consumption requested by patient postoperatively (morphine, 2 mg, iv, each) was recorded. RESULTS: Pain scores were lower in group 1 than in group 2 from 4 to 36 h postoperatively (Mann Whitney rank-sum test). Also, the consumption of supplemental analgesic morphine was lower in group 1 than in group 2 from 6 to 36 h postoperatively. CONCLUSIONS: Intra-articular morphine (3 mg) provided long-lasting analgesia from 6 to 36 h postoperatively and this effect was not due to systemic absorption of morphine. PMID- 8705156 TI - Clinical applications of excitatory amino acid antagonists in pain management. AB - Pain is an important indicator of stress particularly after surgical operation. It is not only a physical and mental suffering but also a main factor for postoperative complications. Relieving of postoperative pain is perquisite for improving outcome and shortening hospital stay. Opioids are still the mainstream for pain relief in common practice. However, opioids are also accompanied by many side effects which limit their usefulness. Therefore, methods to avoid the narcotic-associated side effects are still being searched. Excitatory amino acid receptor antagonists, particularly the NMDA receptor, have been demonstrated to produce satisfactory analgesic effect under certain conditions. This article is an introductory overview of some possible applications of the excitatory amino acid receptor antagonists in future clinical pain management. PMID- 8705157 TI - Long-term propofol infusion and airway management in a patient with Goldenhar's syndrome. AB - A 2-year-old patient of Goldenhar's syndrome received an operation for corneal transplantation. Difficult endotracheal intubation from the congenital anomaly was treated with laryngeal mask airway and pediatric fiberoptic laryngoscope. Long-term propofol infusion (> 10 h) for anaesthetic maintenance in this small child was used with rapid and smooth recovery. PMID- 8705158 TI - Intraoperative somatosensory evoked potentials for localization in excision of recurrent parasagittal meningioma--a case report. AB - Removal of tumors that are located in the motor area is a challenge to neurosurgeons because of the possibility of injuring the motor cortex. We present our experience in excision of a recurrent parasagittal meningioma located in the central area. With the help of somatosensory evoked potentials (SSEP) monitoring, the tumor was removed totally without any neurologic sequela. PMID- 8705160 TI - Advanced practice. PMID- 8705159 TI - Public service private means. PMID- 8705161 TI - Buying nurse education. PMID- 8705162 TI - Shop around for the best education. PMID- 8705163 TI - Supervise to grow. PMID- 8705164 TI - Career into the future. PMID- 8705166 TI - Legal restraint. PMID- 8705165 TI - Health or social care? PMID- 8705167 TI - Education commissioning--a touch of reality. PMID- 8705168 TI - Equal to efficiency. PMID- 8705169 TI - Long term physical complications of battering: an afrocentric intervention of the ancestors. AB - Domestic violence occurs in all strata of American society. This author discusses the prevalence of domestic violence in the African American community; its causes and treatment with specific focus on the role of the ancestors. PMID- 8705170 TI - Oppression's influence on behavior: first hand impressions. AB - This author, who has been in Latvia every year since 1989 to teach, consult, observe health care and hold seminars for nurses, discusses conditions as she has viewed them in Latvia after the demise of Communism and attempts to explain with the use of theory why conditions exist which prevent the full implementation and acceptance of democracy using oppressed group theory. PMID- 8705171 TI - Integrating cultural diversity theoretical concepts into the educational preparation of the advanced practice nurse: the Cultural Diversity Practice Model. AB - Educational preparation of the Advanced Practice Nurse (APN) for the challenging role of providing available and accessible health care to ethnic diverse populations requires that cultural diversity be included in the expanded education. The Cultural Diversity Practice Model is offered for that purpose. PMID- 8705172 TI - Authorship. International Academy of Nursing Editors. AB - There are no binding or universally accepted rules to answer the questions who is an author and in collaborative works, how should the order of authorship be decided? What follows is a response to those questions in the form of a position paper which was endorsed by editors attending the 1993 InANE conference in Edmonton, Alberta, Canada. PMID- 8705173 TI - Fraudulent publication. International Academy of Nursing Editors. AB - The following position paper was prepared by the author and was endorsed by members of the International Academy of Nursing Editors (InANE) on August 20, 1993 at their annual meeting held in Edmonton, Alberta, Canada. PMID- 8705174 TI - Ultrasound diagnosis of portocaval anastomosis in infants--a report of eight cases. AB - We report eight cases of portocaval anastomosis in infants diagnosed by ultrasound. Anatomically we believe this represents continuing patency of the ductus venosus, either as a primary developmental abnormality or secondary to established liver cell failure. PMID- 8705175 TI - Intracranial arterial calcification and ectasia in visual failure. AB - Calcification of the major intracranial arteries is frequently observed on computed tomography (CT), particularly in the carotid siphon, and has been implicated as a causative factor in low tension glaucoma. Ectasia of the carotid siphon has also been postulated as a possible cause of compressive optic neuropathy. 153 patients, in whom high resolution CT of the orbits and parasellar region had been performed for suspected anterior visual pathway pathology, were retrospectively reviewed. The presence and severity of calcification and ectasia of the major intracranial arteries was recorded. There was a high occurrence of carotid siphon calcification, increasing with age, and independent of sex or race. Calcification was very rare in anterior cerebral, middle cerebral and basilar arteries in all age groups. The amount and degree of ectasia also increased with advancing age, and was more frequently observed in carotid and basilar than anterior and middle cerebral arteries. There was no correlation between the degree of calcification or ectasia and ophthalmological or neurological deficit. PMID- 8705176 TI - Posterior pituitary ectopia and hypopituitarism--magnetic resonance appearances of four cases and a review of the literature. AB - In four patients presenting in childhood with varying degrees of hypopituitarism, magnetic resonance imaging (MRI) showed a reduction in size of the normal pituitary fossa contents and an absent or very narrow stalk. A high signal intensity, enhancing area at the base of the stalk, having the appearances and signal characteristics of the posterior pituitary, was seen in each case. We discuss the case histories and MR findings in our patients and review the relevant literature. PMID- 8705177 TI - Focal areas of increased renal tracer uptake on bone scans can mimic metastases in the lower ribs. AB - Focal renal concentration of radioactivity can simulate metastatic deposits in the lower ribs on bone scintigrams. 400 consecutive bone scintigrams were reviewed for focal areas of increased tracer activity in the kidneys. 62 patients (15.5%) had at least one renal hot spot. In 42 of these 62 patients (67.7%) there was an increased focal uptake in the left kidney (18 upper pole, 18 middle pole and six lower pole). 50 out of 62 bone scans (80.7%) showed increased activity in the right kidney (14 upper pole, 32 middle pole and four lower pole). With the incidence of focal areas of activity in the upper poles of the kidneys being 8%, the interpretation of a single focal abnormality projected over the lower ribs should be made with particular care, especially in cancer patients. PMID- 8705178 TI - Application of the principle of optical phase-contrast microscopy to velocity phase-encoded MRI of blood flow in the aorta. AB - A new method of presenting magnetic resonance phase information is described and an example of its application given in the context of velocity phase-encoded MRI of blood flow in the aorta. The method takes as its starting point Zernike's technique of phase contrast microscopy. It exploits the parallel between the transform plane in Fourier optics and kappa-space in MRI. In the example described two datasets are acquired, one with and the other without velocity encoding, as in conventional phase-encoded velocity imaging. A dataset is formed which is corrected for unwanted phase variations caused by static field inhomogeneity. The method then effectively combines phase and magnitude information into a single image. The technique is complementary to existing methods of displaying phase information. PMID- 8705179 TI - Assessment of target recovery after TV camera saturation in X-ray fluoroscopy using a personal computer with frame-grabber. AB - X-ray fluoroscopy images were digitized with a personal computer and frame grabber system. This system was used to measure the recovery from saturation of the target of a TV camera by calculating the time at which contrast falls to 50% (C 1/2). This measurement was performed on images of: (a) a rotating lead disk with a slit at its side and (b) a square hole in a thick lead sheet. These images represented the moving (MOTR) and stationary (SOTR) object target recovery. The SOTR method produced much higher C 1/2 values in two mobile fluoroscopy systems used for cardiac catheterizations where waveforms permitted the application of both methods. The SOTR method was also applied to three additional fluoroscopy systems. It was found that, contrary to expectations, C 1/2 increased with exposure rate in all five machines. Target recovery characteristics of the machines tested, represented by the gradient of C 1/2 against exposure rate, were found to increase with inherent noise level measured in the vidicon camera target. The set-up used here for measuring target recovery proved to work quickly and reproducibly for both SOTR and MOTR. If initial values are calculated then the condition of any fluoroscopy system may easily be followed irrespective of the method used. PMID- 8705180 TI - Patient doses in bone mineral densitometry. AB - Procedures were developed to compute effective dose equivalent (H(E)) values for patients undergoing bone mineral densitometry (BMD) measurements such as dual energy X-ray absorptiometry (DEXA) and quantitative computed tomography (QCT). Representative values of H(E) were determined for patients undergoing each type of BMD procedure. Typical DEXA measurements are associated with an H(E) of about 2.5 microSv. For QCT, the values of H(E) are 300 microSv for single energy techniques and 1 mSv for dual energy techniques, respectively. By comparison, a single well collimated abdominal X-ray examination often taken in conjunction with DEXA studies results in an H(E) of about 100 microSv. BMD patient doses are at the lower end of the exposure range encountered in diagnostic radiology. As a result, radiation dose is not a primary factor in choosing the method for BMD determination. PMID- 8705181 TI - Effective doses for coronary angiography. AB - A detailed study of dose data has been made for 100 coronary angiography examinations performed on a digital X-ray unit. Dose-area product (DAP) data have been analysed in terms of fluoroscopy and radiography for different projections. Projections patterns were similar for all cardiologists studied and the contribution to DAP from fluoroscopy was 32%. Data for 10 patients, selected because the percentage contributions to DAP from fluoroscopy and radiography were similar to the mean for the group, were used to derive conversion factors from DAP to effective dose. The mean DAP was 14 Gy cm2 and the effective dose 3.1 mSv. Conversion factors, based on the same protocol, were derived for a range of tube potentials and filtrations for use in estimation of effective doses with other units. PMID- 8705182 TI - Patient doses from CT in New Zealand and a simple method for estimating effective dose. AB - A 1992 survey of doses to patients from CT scanning in New Zealand found that CT contributes 81 microSv per capita per annum, representing about 17% of the collective dose from medical irradiation. Because of the significance of CT, estimates of patient dose are now included in the National Radiation Laboratory (NRL) routine surveys of CT scanners. For some models of CT scanner, normalized organ dose data from Monte Carlo studies are available, enabling the calculation of the effective dose. For scanners for which there are no normalized organ dose data, it is shown that the computed tomography dose index (CTDI), determined at 1 cm depth in Perspex phantoms and normalized for mAs, may be used to estimate the effective dose. Estimates of effective dose derived from the CTDI may then be used to formulate reference dose levels, against which individual scanner doses may be compared. PMID- 8705183 TI - A comparison of doses and techniques between specialist and non-specialist centres in the diagnostic X-ray imaging of children. AB - A dosimetric survey of 14 routine X-ray examinations in children was carried out between 1993 and 1995. Two children's hospitals and four general hospitals took part in the survey which involved the calculation and measurement of nearly 3000 doses. Entrance surface doses (ESD) were calculated from exposure factors for radiographic procedures, and dose-area products (DAP) were recorded for both radiographic and fluoroscopic procedures. Doses were in good agreement with earlier studies, but for some procedures were significantly lower than those reported from other European countries. The main dose influencing factors for radiographic procedures were found to be the speed of the film-screen system and the use of an antiscatter grid. For the main head/trunk examinations, specialist centres often delivered higher doses to the younger children as a result of widespread use of a grid. In fluoroscopy, where the main dose influencing factors were the use of a grid and the dose rate dependence of the image intensifier, the children's hospitals consistently delivered significantly lower doses. Both ESDs and DAPs were found to increase with patient age for the main head/trunk examinations, although in some cases (AP/PA chest) this relationship was weak. The dependence of dose on age necessitates the subdivision of the paediatric sample into a number of age categories. It is suggested that all authors use the same age groupings. PMID- 8705184 TI - Assessment of osteopenia from spine radiographs using two different methods: the Chingford Study. AB - Two methods for diagnosing radiological osteopenia in thoracic (TS) and lumbar (LS) spine radiographs were assessed: a subjective conventional method (A) and a semiquantitative method (B), by comparing them with bone mineral density (BMD) measured by dual energy X-ray absorptiometry (DEXA), in a population of "normal" women aged 45-70 years (n = 818). For both methods there was good intraobserver and interobserver reproducibility. BMDs were significantly lower with increasing radiological osteopenia grades (p < 0.001), and remained lower after adjustment for age and body mass index (p < 0.01). The proportion of subjects with DEXA defined osteoporosis rose with increasing radiological osteopenia grades for both methods. The worst osteopenia categories identified 29.7-55.3% of women with DEXA defined osteoporosis, compared with 6.1-11.7% in the "normal" categories. Both methods, however, showed a large degree of overlap of BMDs between the various radiological osteopenia grades. The sensitivity and specificity of method A in diagnosing osteoporosis were 45.3% and 78.4%, respectively, for the TS and 19.0% and 94.3%, respectively, for the LS. For method B the sensitivities and specificities were 8.8% and 96.1%, respectively (TS), and 10.2% and 95.6%, respectively (LS). Although both methods have poor sensitivities, "definite" or "high" grade osteopenia should be an indication for bone densitometry. The high specificities suggest that a "normal" (no osteopenia) X-ray is unlikely to have a significantly low BMD. PMID- 8705185 TI - The influence of the number of fractions and bi-exponential repair kinetics on biological equivalence in pulsed brachytherapy. AB - A linear-quadratic radiobiological model incorporating single or bi-exponential repair kinetics has been used to show the following and other features when a continuous low dose rate (CLDR) 70 Gy/140 h brachytherapy protocol is replaced by a radiobiologically equivalent pulsed dose rate (PDR) system using 140 fractions for reasons of dosage homogeneity. (1) For equivalent effects in late-reacting tissues, the PDR dose (at 5 or 0.05 Gy min-1) x 1 h intervals needs to be reduced by up to only 3%. Progressively further reductions in dose are required when fewer larger fractions are used. (2) When equivalence using pulsed doses is achieved for one normal tissue type, and extrapolated response doses (ERD) are calculated for other tissue types in the irradiated volume, values of the ERD remain within 5% of each other using the above PDR protocol and associated parameters. (3) For tumours with alpha/beta = 10 Gy and a single repair halftime of 0.1-1.0 h, there is no significant loss of therapeutic benefit using the PDR protocol equivalenced for late normal tissue reactions. The strategy of replacing an LDR boost protocol of about 24 Gy by a PDR protocol gives similar levels to the 70 Gy PDR protocol for the expected percentage increase in the biological dose to normal tissues (due to the PDR protocol alone). These calculations also highlight the importance of the values assumed for the conventional alpha/beta ratio and the repair kinetics when estimating equivalent PDR protocols. The use of an inappropriate radiobiological parameterization will lead to erroneous conclusions with the potential to advocate PDR protocols which will, in practice, lead to an increase in late complications. PMID- 8705186 TI - Short communication: a system for remote monitoring of a hospital linear accelerator. AB - Linear accelerators are complex machines with many parameters affecting the quality of the treatment beam delivered. A high level of technical support is required but this can be difficult to achieve if the linear accelerator is at a centre distant from a medical physics department. This paper describes a solution to this problem whereby a newly installed linear accelerator at the Royal Shrewsbury Hospital is remotely monitored from the Royal Hospital, Wolverhampton. The system enables run-up procedures to be completed by guiding radiographers at Shrewsbury through a series of steps. It consists of electronic hardware connected to the linear accelerator, under the control of a computer. The machine parameters are read and tested against preset tolerances. The monitoring system has been installed and is in routine use. The benefits have been shown to be: a saving in staff time and travel costs, the satisfactory completion of run-up procedures, a higher level of efficiency in the medical physics service provided and effective quality control. Finally, some future developments are presented. These include the development of a real time remote monitoring system that constantly monitors the linear accelerator, allowing the Medical Physics Department access to the machine parameters at any time. PMID- 8705187 TI - Case report: Pneumocystis carinii pneumonia presenting as a solitary cavitating lung lesion in non-HIV immunosuppressed patients. AB - Pneumocystis carinii is one of the most common pulmonary opportunistic infections in immunodeficient patients. It has a classical chest radiographic appearance of bilateral interstitial infiltrates. Recently, cystic or cavitating lesions have been reported in HIV patients with Pneumocystis carinii pneumonia. We report two cases of non-AIDS immunosuppressed patients with the unusual presentation of a solitary cavity in the lung. PMID- 8705188 TI - Case report: paravesical suture granuloma resembling bladder carcinoma on CT scanning. AB - Paravesical granuloma formation is a rare complication of inguinal herniorrhaphy. Infected non-absorbable suture material is the usual cause. We present a case of paravesical granuloma that was initially thought to be bladder carcinoma both clinically and on CT scanning. The patient presented with urinary symptoms. Repeated biopsies revealed only reactive changes with no evidence of malignancy. CT showed a mass which appeared to arise from the anterior aspect of the bladder with a central area of low attenuation. The diagnosis was established at cystoscopy with deep resection of the mass, when two retained silk stitches with surrounding pus and granulation tissue was found. The diagnosis of paravesical suture granuloma (PSG) should be considered in patients with suprapubic symptoms, no haematuria, an atypical mass and a history of herniorrhaphy, which may have been many years prior to the time of presentation. PMID- 8705189 TI - Case of the month: that's torn it! PMID- 8705190 TI - Late aggressive meningioma following prophylactic cranial irradiation for acute lymphoblastic leukaemia. PMID- 8705191 TI - A burning question. PMID- 8705192 TI - Magnetic resonance imaging-guided pre-operative breast localization using a "freehand technique". PMID- 8705193 TI - Patient good versus patient good. PMID- 8705194 TI - Application of a simplified comparative genomic hybridization technique to screen for gene amplification in pediatric solid tumors. AB - Conventional cytogenetic analysis of solid tumors is technically very demanding and requires a large number of viable cells. The technique of comparative genomic hybridization (CGH) circumvents these difficulties and has been shown to be particularly useful for identifying new gene amplifications. We have simplified the CGH technique for the detection of amplifications by utilizing a single labeling approach in which labeled tumor DNA is mixed with unlabeled normal human DNA and hybridized to normal metaphases on a slide. To examine the consistency and sensitivity of the method, initial experiments were performed using a retinoblastoma (RB) cell line and five pediatric solid tumors known to contain an amplification. The technique was easy to use and sensitive enough to detect low level amplifications. The RB cell line showed reproducible signals at 2p24, indicative of amplified sequences, on both homologues in 95% of the metaphases (> 30) examined. Amplifications of the MYCN gene (2p24) were detected in three alveolar rhabdomyosarcomas and one medulloblastoma. CGH was then applied to six tumors in a prospective fashion, before data about specific gene amplification were available. In two, amplification of the MDM2 gene (12q13-14) was identified using CGH and later confirmed by Southern blot analysis. Four tumors negative for MDM2 and MYCN amplifications by CGH analysis were also negative by Southern blot analysis. Gene amplification as low as fourfold was detected in one tumor and the overall pattern of gene amplification detected by CGH in these tumors was not complex, involving just one amplification site for each case. Therefore, this simplified CGH technique is suitable for routine screening of pediatric solid tumors for amplifications when genetic studies are important but sample sizes are small and dividing cells are infrequent or unavailable. PMID- 8705195 TI - Comparison between flow cytometric and cytogenetic tumor cell DNA content using a simple flow sample preparation method: ploidy comparison of 86 fresh pediatric tumors. AB - A comparison of tumor ploidy by flow cytometry (FL) and cytogenetics (CYG) was made in 86 fresh pediatric solid (n = 47) and hematopoietic (n = 39) tumors using simple sampling and semiautomated proprietary FL preparation methods and defined histogram interpretive criteria. Tumor karyotypes with 44-48 chromosomes were regarded as CYG diploid and other chromosomal complements CYG aneuploid for comparison purposes. Five histograms were uninterpretable and nine cases failed to produce 15 or more metaphases for karyotyping. Mean G0/G1 peak coefficients of variation of all 86 cases were 2.7 and 3.0 for the diploid and aneuploid populations, respectively. Of the 72 eligible cases, 41 were concordant diploid and 16 concordant aneuploid with an overall concordance of 79%. The DNA index and karyotypic index correlation coefficient was 0.92 for the 16 concordant aneuploid cases. Analysis of the 15 discordant cases highlights the limitations of both methods and of the histogram interpretive criteria and indicates that FL is probably more sensitive for detection of tumor aneuploidy as defined and detected by these methods. PMID- 8705196 TI - Castleman's disease in children: the experience of a children's hospital in Africa. AB - As the incidence of childhood Castleman's disease (CD) is reported to be rare in Western communities, the recent occurrence of three cases in non-HIV infected prepubertal children stimulated a 10-year (1983-1993) retrospective investigation into the conditions at a children's hospital that serves a predominantly developing community. Retrospective histologic examination revealed six cases of CD among children 3-11 years of age and another five probable cases having histological features of CD without adequate clinical and laboratory data. Three CD cases were associated with malignancy (Kaposi's sarcoma in two and a gastric B cell lymphoma in the other). Two CD cases were plasma cell type, one hyaline vascular, and three intermediate. One child with recurrent plasma cell-type features in a groin node had a Kaposi's sarcoma in the thigh. The five probable cases of CD all had intermediate histological features. Four of the 11 cases being reported came from a geographic locality that provides about 3% of the laboratory's case material and these included both cases that developed Kaposi's sarcoma. All patients who developed malignancy had the plasma cell variant. As all cases have been identified in developing communities, where the disease may often be misdiagnosed as nonspecific reactive lymphadenopathy, and there is an evident association with malignancy, it is recommended that children in these areas with the plasma cell variant of CD be closely monitored for subsequent malignancy. PMID- 8705197 TI - Stability of twenty-five analytes in human serum at 22 degrees C, 4 degrees C, and -20 degrees C. AB - The stability of 25 analytes from serum of healthy donors was determined at room temperature, 4 degrees C, and -20 degrees C over 48 h, 14 days, and 4 months, respectively. Glucose, blood urea nitrogen, sodium, potassium, chloride, creatinine, calcium, phosphorus, uric acid, total protein, albumin, triglycerides, lipase, total creatine kinase, gamma-glutamyltransferase, iron, magnesium, and cholesterol were stable at all three temperatures for the specified times. Carbon dioxide, aspartate and alanine aminotransferases, lactate dehydrogenase, amylase, alkaline phosphatase, and high-density lipoprotein cholesterol demonstrated some loss over time. Proper storage temperatures and times must be considered for these analytes if measurement is not to take place immediately after specimen collection. PMID- 8705198 TI - In situ hybridization of human erythropoietin in pre- and postnatal kidneys. AB - Due to the transient transcription of the gene of erythropoietin (EPO) in response to hypoxic stimuli, localization within the cell of origin in humans has not been possible. Under acutely induced transcription of the EPO gene, through severe bleeding (hematocrit < 10%), EPO transcripts were detected within renal interstitial cells in rodents. However, by immunohistochemistry or tissue culture, human tubular epithelial cells or glomerular mesangium have been shown to produce EPO, suggesting a species difference. Aiming to investigate the precise location of EPO in intact human kidneys, 38 fetal, infant, and adult kidneys were examined by in situ hybridization. Kidneys from patients with conditions predisposing to severe hypoxia and/or induced polycythemia were utilized, because under these conditions active EPO mRNA synthesis is expected. We found specific EPO mRNA transcripts within a small population of cortical interstitial cells of near-term or postnatal kidneys. PMID- 8705199 TI - Cystic dysplasia of the testis: a report of three cases and review of the literature. AB - We describe the clinical and pathologic findings in three cases of cystic dysplasia of the testis and compare our findings to others in the English literature. The condition is characterized by dilatation of the rete testis with or without similarly affected adjacent structures. It is important to recognize this lesion for two reasons. First, it is associated with a high frequency of ipsilateral renal anomalies. Second, it is important to consider this condition in the differential diagnosis of cystic testicular mass in a neonate or a child. The management of this condition is quite different from the management of other testicular masses such as a hydrocele, testicular germ cell tumor, or infiltrative disorder (e.g., lymphoma, inflammation, torsion, or hematoma). Testicular cystic dysplasia is benign and the lesion can be removed surgically, leaving the rest of the testis intact. A discussion of the lesion's possible pathogenesis is also presented. PMID- 8705200 TI - Bilateral ulnar agenesis: case report and review of the literature. AB - Bilateral ulnar agenesis is a rare abnormality. A total of 36 cases are analyzed: 35 of these are documented in the literature and 1 stillborn male is presented in this study. Most patients had one of the three conditions: Al-Awadi/Raas Rothschild syndrome, syndrome of ulnar aplasia with split hand/split foot deformity, or the Brachmann-de Lange syndrome. Fifty percent of all cases with bilateral ulnar agenesis were associated with lower limb defects and these cases, for the most part, also belonged to the aforementioned syndromes. Nonskeletal, internal organ malformations were identified in 34% of all patients. Nine patients presented with isolated bilateral ulnar agenesis. The Al-Awadi/Raas Rothschild syndrome and the split hand/split foot deformity are heritable disorders. There was no evidence for genetic etiology in most of the other cases. Bilateral ulnar agenesis in our fetus was part of the Brachmann-de Lange syndrome with associated cardiac defect, diaphragmatic hernia, and umbilical artery agenesis. PMID- 8705201 TI - Familial renal tubular dysgenesis: a disorder not isolated to proximal convoluted tubules. AB - The autopsy findings of a newborn with renal tubular dysgenesis, born to first cousins of Moslim Arab descent, are described. Hypocalvaria and hyperflexible joints were noted in addition to the renal lesion. A microdissection study demonstrated marked shortening of all the nephron segments from the glomeruli to the collecting tubules, rather than an isolated abnormality of the proximal convoluted tubules. PMID- 8705203 TI - Monozygotic twins concordant for congenital neuroblastoma: case report and review of the literature. AB - To date, five sets of monozygotic twins concordant for neuroblastoma have appeared in the literature. Review of the clinical information available for these cases suggests that they represent congenital, versus acquired, diseases in both twins from each twin pair. The question arises, then, whether some or all sets of twins represent simultaneous-onset malignancy of both twins within a twin pair or whether metastasis via placental anastomoses from one twin with congenital disease to the cotwin occurs. This report includes a sixth set of monozygotic twins concordant for congenital neuroblastoma. From analysis of the clinical data from all cases, it appears that two of six twin pairs may represent simultaneous-onset tumors in each twin from a twin pair, and two of the remaining four twin pairs, including the present case, may represent placental metastases from one twin with congenital neuroblastoma to the other. In the remaining two twin pairs insufficient data are available upon which to draw any conclusions. Now that molecular and genetic methodology is available to characterize neuroblastomas, these techniques may be utilized in future cases of monozygotic twins concordant for congenital neuroblastoma, to help clarify whether the neoplasms represent simultaneous primary tumors versus metastatic spread from one twin to another. PMID- 8705204 TI - Partial deletion of chromosome 6p: autopsy findings in a premature infant and review of the literature. AB - Autopsy findings for a premature infant with terminal deletion of 6p [46, XY, del(6)p23] are reported. The infant had multiple dysmorphic features, cleft lip and palate, abnormal external genitalia, diverticulum of right ventricle, and hydrocephalus with cerebellar hypoplasia, with absent corpus callosum and cerebellar vermis. PMID- 8705202 TI - Adrenal gland leiomyoma in a child with acquired immunodeficiency syndrome. AB - An adrenal gland leiomyoma was incidentally found at autopsy in a 2-year-old boy with acquired immunodeficiency syndrome (AIDS). Smooth muscle neoplasms have been described in pediatric AIDS and affect mainly the tracheobronchopulmonary, gastrointestinal, and hepatobiliary systems. An association between Epstein-Barr virus (EBV) and smooth muscle tumors in patients with AIDS has been described. This case is the first leiomyoma to be reported in the adrenal gland of a child with AIDS. EBV genome was demonstrated in the tumor by polymerase chain reaction. PMID- 8705205 TI - Congenital CNS primitive neuroectodermal tumor: case report and review of the literature. AB - A 30-week gestational age male fetus was found to have a congenital neoplasm involving the posterior cranial fossa, identified by fetal ultrasound examination in utero. Histological and immunohistochemical examination confirmed a diagnosis of primitive neuroectodermal tumor (PNET) of the posterior fossa, also referred to as medulloblastoma. Although PNETs have been well documented, there are relatively few reports of these occurring as congenital neoplasms. We present a case of an in utero, congenital PNET with a review of the literature and discussion of the criteria defining congenital tumors. PMID- 8705206 TI - Quality assurance in pediatric anatomic pathology: the Society for Pediatric Pathology slide survey program. AB - The Society for Pediatric Pathology initiated an external slide survey program in 1989 and has produced yearly challenges of 12 autopsy and surgical pathology glass slides. The history and operational difficulties are reviewed. As the program matures, it presents new challenges in the area of proficiency testing and case acquisition. PMID- 8705207 TI - New technology in urology: why assessment needs to be more scientific. PMID- 8705208 TI - Anti-androgens in the treatment of prostate cancer. PMID- 8705209 TI - A new technique for detrusor biopsy and its applicability in the ultrastructural study and diagnosis of voiding dysfunction. AB - OBJECTIVE: To determine the clinical applicability of ultrasound-guided detrusor biopsy from the anterior bladder wall and to assess whether it is as representative and as sufficient for determining detrusor ultrastructure as the traditional transurethral biopsy from the lateral bladder wall. MATERIALS AND METHODS: The detrusor structure in 22 biopsies. 11 obtained transabdominally and 11 transurethrally from 11 patients, was evaluated without knowledge of the biopsy method by light and electron microscopy, including morphometric analysis. In addition, several specimens from each of three bladders were evaluated, also 'blindly', for ultrastructural diagnosis of the detrusor in an independent current study of voiding dysfunction in geriatric patients. RESULTS: There were no differences in intercellular distances and cell: nucleus ratios between transabdominal and transurethral biopsies in eight of the 11 patients. Furthermore, ultrasound-guided transabdominal biopsies were as representative of the detrusor as were transurethral biopsies. CONCLUSION: The transabdominal approach is recommended as an easy, relatively inexpensive and efficient method of obtaining biopsies of the detrusor for study of its structure in voiding dysfunction. On the other hand, the observations and diagnoses made by 'blind' qualitative study of several specimens from the same bladder were identical. This, together with the similarity of detrusor structure in the transabdominal and transurethral biopsies, strongly supports the idea that such structure is relatively uniform throughout the entire bladder wall. PMID- 8705210 TI - Urodynamic changes during therapeutic intravesical instillations of capsaicin. AB - OBJECTIVE: To describe the technique for and urodynamic changes during therapeutic instillations of intravesical capsaicin in patients with detrusor hyper-reflexia. PATIENTS AND METHODS: Ninety intravesical instillations of capsaicin were performed as a therapeutic procedure in 30 patients; 21 patients had various causes of non-traumatic spinal cord disease, five patients were very severely neurologically impaired and were bed-bound with an indwelling catheter, and four were neurologically normal. Simultaneous cystometry was performed in 25 patients during the instillation of capsaicin; 100 mL of 1 or 2 mmol/L capsaicin in 30% ethanol/saline was instilled into the bladder for 30 min and two patients received 30% ethanol/saline only. The last 56 capsaicin treatments were preceded by the instillation of 40 mL of 2% lignocaine for 20 min. Detrusor hyper-reflexia was decreased and urinary continence improved for 3-6 months after a single instillation; the instillation was then repeated. Two patients who received only ethanol/saline showed no clinical or urodynamic improvement. RESULTS: The treatment was not abandoned in any patient due to discomfort and there were no short- or medium-term complications. All patients with spinal cord disease and phasic detrusor hyper-reflexia had similar, frequent and repetitive detrusor contractions during the instillation of capsaicin. These acute reactive contractions did not occur in the neurologically normal patients. Similarly, the instillation of intravesical lignocaine only caused no phasic detrusor contractions. Intravesical lignocaine instillation before capsaicin markedly reduced and sometimes abolished the detrusor overactivity and lessened the discomfort for the patients. The instillation of lignocaine before capsaicin did not alter the benefit from each instillation of intravesical capsaicin. CONCLUSION: A method has been developed for administering capsaicin intravesically which diminishes discomfort for the patient and in the short- and medium-term is free of complications. The study also provides functional evidence of the role of capsaicin-sensitive afferents in phasic detrusor hyper-reflexia due to spinal cord disease. PMID- 8705211 TI - Altered electrical properties of bladder smooth muscle in streptozotocin-induced diabetic rats. AB - OBJECTIVE: To examine the alterations in the electrical properties of bladder smooth muscle in rats with diabetes mellitus (DM) induced by streptozotocin. MATERIALS AND METHODS: The study comprised 34 control rats and 20 with DM induced experimentally by injection with streptozotocin. At 10-15 weeks after the induction of DM, control and treated rats were killed, the bladder removed and the electrical responses of membranes from the detrusor smooth muscle recorded using intracellular microelectrodes. RESULTS: The resting membrane potential of the smooth muscle remained unaltered by DM but the frequency of spontaneous spike discharges sensitive to nicardipine was decreased. Transmural nerve stimulation elicited a purinergic excitatory junction potential (EJP) in muscles from both types of rat, but the threshold intensity of stimulation required to evoke an EJP was higher in muscle from the diabetic rat. Furthermore, EJPs large enough to trigger a nicardipine-sensitive spike potential could be elicited easily in muscles from the control rat, but not in muscles from diabetic rats. Stimulation of muscarinic receptors with exogenous acetylcholine (ACh) caused greater depolarization in muscle from diabetic rats. Stimulation of purinergic receptors with alpha, beta-methylene ATP caused a similar depolarization in both groups of rats. Application of potassium-free solution or ouabain depolarized the membrane by about 9 mV or 5 mV, respectively, in muscles from both groups of rat. Removal of potassium-free solution produced an ouabain-sensitive transient hyperpolarization which was larger in muscle from control rats, indicating that the potency of the Na-K pump was weakened in rats with DM. CONCLUSION: In the detrusor smooth muscle of rats with DM, there were fewer spontaneous spike discharges, supersensitivity of post-junctional muscarinic receptors, reduced potency of the post-junctional Na-K pump and a decrease in the release of neurotransmitter, possibly due to the impairment of prejunctional activity. PMID- 8705212 TI - The Bristol Female Lower Urinary Tract Symptoms questionnaire: development and psychometric testing. AB - OBJECTIVE: To develop a questionnaire that is sensitive to changes in the symptomatology of the female lower urinary tract, particularly urinary incontinence, providing an instrument that can characterize symptom severity, impact on quality of life and evaluate treatment outcome. PATIENTS, SUBJECTS AND METHODS: Items covering as wide a range of urinary symptoms as possible were devised after consultation with clinicians and a health scientist, a literature review and discussion with patients. Additional items assessed the degree of 'bother' that symptoms were causing. Eighty-five women with clinical symptoms attending for urodynamic assessment and 20 women with none were asked to self complete the questionnaire. The instrument's validity was assessed by interviewing patients and measuring levels of missing data, comparing symptom scores between clinical and non-clinical populations and comparison with frequency/volume charts and data from pad tests. The instrument's reliability was assessed by measuring both internal consistency and stability, using a 2-week test-retest analysis. RESULTS: The questionnaire was completed by the patients with a mean of only 2% of items missing; most questions were easily understood. Construct validity was good, with the instrument easily differentiating clinical and non-clinical populations. Criterion validity, as tested against frequency/volume charts and pad-test data, was acceptable, with Kappa coefficients of 0.29-0.79 for frequency/volume data and Spearman rank correlations of 0.50-0.97 and 0.31-0.67 for frequency/volume and pad-test data, respectively. The reliability of the instrument was good; a Cronbach's alpha of 0.78 indicated that the symptom questions had high internal consistency, while stability was excellent, with 78% of symptoms and problems answered identically on two occasions, and Spearman rank correlations of 0.86 and 0.90, respectively. CONCLUSION: The instrument has good psychometric validity and reliability. The stability demonstrated at baseline and the ability to differentiate clearly between community and clinical populations suggest that it should be ideal for measuring changes following therapeutic intervention. The addition of life-impact items and a 'bother' factor may provide the opportunity to identify those women who wish treatment for their symptoms; this dimension requires further exploration. PMID- 8705213 TI - The value of multiple free-flow studies in men with lower urinary tract symptoms. AB - OBJECTIVE: To assess the variability of free-flow studies in men presenting with lower urinary tract symptoms (LUTS) suggestive of benign prostatic obstruction (BPO) and to determine the sensitivity, specificity and predictive values of consecutive measurements of maximum flow rate for the presence of bladder outlet obstruction (BOO) at several threshold values. PATIENTS AND METHODS: The value of multiple free-flow studies was assessed in 165 men presenting with LUTS suggestive of BPO. Each patient was requested to void four times into a uroflowmeter and the voided volume and post-void residual urine volume (PVR) were also measured. The variability of the maximum flow rate (Qmax), voided volume and PVR between consecutive voids was assessed. In addition, the sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of various thresholds of Qmax for the presence of BOO were calculated for each consecutive void. RESULTS: The mean Qmax on void 1 was 10.2 mL/s and the mean maximum value for Qmax between voids 1 and 2 was 12.5 mL/s. For voids 1, 2 and 3, the mean maximum Qmax was 13.9 mL/s and for voids 1 to 4 it was 15.2 mL/s. There were no significant changes in PVR among any of these voids. There was a statistically significant, although small, decrease in voided volume between voids 1 to 3 and voids 1 to 4. The specificity and PPV of Qmax for BOO increased with each subsequent void, such that using a threshold value for Qmax of 10 mL/s on the fourth void, the specificity and PPV for BOO were 96% and 93%, respectively. CONCLUSIONS: There was a significant increase in Qmax with each successive void when men with LUTS suggestive of BPO performed multiple free-flow measurements and consequently, single free-flow measurements substantially underestimated the maximum Qmax that these patients achieved. The specificity and PPV of Qmax for BOO can be improved considerably by performing multiple free-flow studies and by carefully selecting an appropriate threshold value (although whether pressure-flow studies are unnecessary will depend on what level of specificity and PPV is deemed acceptable in clinical practice). These findings should be considered if free-flow studies are to be used as the basis for deciding the clinical management of men with LUTS and may be particularly useful for urologists with limited facilities for pressure-flow studies. PMID- 8705214 TI - Determination and reversal of resistance to epirubicin intravesical chemotherapy. A flow cytometric model. AB - OBJECTIVE: To develop a method of determining the characteristics of epirubicin resistance and to study the reversal of such resistance in the intravesical treatment of superficial bladder cancer, using sensitive and resistant derivatives of a bladder cancer cell line in vitro. MATERIALS AND METHODS: Epirubicin fluorescence and flow cytometry were used to measure the intracellular levels of epirubicin in both sensitive and resistant live cultured bladder tumour cells, with and without different doses of the resistance-reversing agent verapamil. RESULTS: There was a reliable, highly significant and consistent difference in intracellular epirubicin concentration between the resistant and sensitive bladder tumour cells. In addition, it was possible to substantially reverse the features of resistant cell subline with additional verapamil. CONCLUSION: Application of this assay to clinical specimens should allow better targeting of epirubicin intravesical chemotherapy and avoid the premature termination of such treatment in patients whose tumours remain sensitive to this agent. Furthermore, the addition of verapamil to intravesical epirubicin may permit effective treatment of those patients whose tumours have inherent or acquired resistance to epirubicin. PMID- 8705215 TI - Determination and reversal of resistance to epirubicin intravesical chemotherapy. A confocal imaging study. AB - OBJECTIVE: To evaluate the use of confocal microscopy in the study of resistance to epirubicin and to determine the effect of temperature, viability and a resistance-reversing agent on the intracellular distribution of this drug in sensitive and resistant derivatives of a superficial bladder cancer cell line. MATERIALS AND METHODS: Viable and non-viable adherent cells were incubated in epirubicin solutions under various conditions. After incubation, the distribution of intracellular epirubicin fluorescence was visualized using confocal microscopy and a x50 water-immersion lens. RESULTS: There was a striking and consistent difference between resistant and sensitive cells in the intracellular distribution of the drug. In addition to having greater overall levels of epirubicin fluorescence, sensitive cells accumulated epirubicin predominantly in the nucleus. Epirubicin fluorescence in resistant cells was cytoplasmic and granular in appearance. When incubated at 0 degrees C, both cell lines showed no nuclear uptake and thus resembled resistant cells at 37 degrees C. However, dead cells rapidly acquired brightly fluorescent nuclei. The resistance-reversing agent verapamil appeared to cause reversion of the resistant to the sensitive phenotype. CONCLUSION: Confocal microscopy allows epirubicin-sensitive and resistant cultured tumour cells to be differentiated reliably and provides information about the mechanisms of action of, and resistance to, epirubicin. Applying this technique to clinical specimens should enable patients who have the resistant phenotype to be detected and the efficacy of intravesical resistance reversing agents to be evaluated in such cases. PMID- 8705217 TI - Video-resection does not stop corneal contamination during transurethral resection of the prostate. AB - OBJECTIVES: To determine the efficacy of video-resection systems in reducing the risk of corneal contamination during transurethral resection of the prostate (TURP) and thus in decreasing the possibility of transmission of blood-borne diseases from the patient. MATERIALS AND METHODS: After performing a TURP using a video camera system in 30 patients, the number of potential corneal droplets was measured for one operator wearing a pair of spectacles and, for a second operator, over an identically sized area of a standard disposable plastic eyeshield. RESULTS: In 20 (67%) of cases, droplets were observed on the eye protection, correlating with potential corneal contamination. Splashes were also seen on the face-mask of the eyeshield. CONCLUSION: Urologists should use eye and face protection when performing TURP using a video system. PMID- 8705216 TI - The benefits of a shared-care prostate clinic. AB - OBJECTIVE: To establish a hospital based shared-care clinic to investigate and manage benign prostatic hyperplasia (BPH) with general practitioners (GPs). PATIENTS AND METHODS: During one year, 330 patients referred with suspected prostatic obstruction were investigated in an outreach clinic in a rural cottage hospital by urology department nurses according to a protocol. After this, they were referred directly back to their GPs with recommendations for their management or seen in the urologist's clinic. A questionnaire was completed by the GPs to assess their satisfaction with and attitudes to the clinic. RESULTS: One-third of the patients were referred directly back to their GP, a third were seen routinely and a third seen urgently in the urologist's clinic, usually because a prostate-specific antigen assay indicated the possibility of latent prostatic cancer. A survey confirmed that GP support for the clinic was unanimous whilst patients were reassured by the thoroughness and sensitivity of the clinic's nursing staff. CONCLUSION: The clinic reduced the workload of the GPs and urologists whilst providing a speedy and comprehensive assessment of patients presenting with suspected prostatic obstruction. PMID- 8705218 TI - Multiple use of fibres in the visual laser ablation of the prostate. AB - OBJECTIVE: To investigate the possibility of re-using laser fibres for the visual laser ablation of the prostate (VLAP). PATIENTS AND METHODS: Thirty-four patients, each with a mean maximum urinary flow rate of < 14.0 mL/s and an American Urological Association symptom score of > 19, were evaluated. All underwent VLAP: laser energy, generated by a Neodymium:YAG system, was delivered with a side-firing fibre. The laser generator was fitted with a custom-built power meter that measured the energy delivered by the fibre. Fibres were re-used until the energy output fell below 60 W despite increases in laser generator power. Each patient was followed for at least 12 months. RESULTS: A total of seven fibres was used to treat the 34 patients; between one and nine patients were treated with each fibre. The outcome obtained with the first use of a fibre compared well with those obtained with each subsequent use. After 12 months, 25 patients (74%) expressed satisfaction with the results of their operation. Five patients (15%) had urinary tract infections. As each fibre costs about pound 650, multiple use saved a mean of pound 516 for each patient treated. CONCLUSION: This evaluation implies that there is no justification for the single use of laser fibres on the basis of efficacy, provided that a power meter is used to monitor the efficiency of the delivery device. PMID- 8705219 TI - Local staging of prostate cancer by endorectal MRI using fast spin-echo sequences: prospective correlation with pathological findings after radical prostatectomy. AB - OBJECTIVE: To evaluate the accuracy of endorectal magnetic resonance imaging (MRI) with fast spin-echo sequences in the local staging of clinically localized prostate cancer. PATIENTS AND METHODS: Seventy-one patients with a clinical T1 (18 patients) or T2 tumour (53 patients) underwent endorectal MRI 2-12 weeks before radical prostatectomy. Extraprostatic tumour was diagnosed if MRI showed signs of capsular penetration and/or invasion of the seminal vesicle and/or distal urethra or bladder neck. If the pathological examination showed a single positive margin with no periprostatic tissue, the tumour was classified as indeterminate and not as a T3 tumour. RESULTS: Of the 25 cases of capsular penetration. MRI correctly identified 10 (sensitivity 42%, specificity 100%). Of the 14 cases with seminal vesicle invasion, MRI correctly identified six (sensitivity 43%, specificity 100%), but showed other signs of extraprostatic tumour spread in seven of the eight unidentified cases. Overall, MRI identified 16 of the 30 patients (53%) with occult extraprostatic spread of tumour; there was only one false-positive result. The sensitivity, specificity and accuracy of MRI were 53, 96 and 74%, respectively. CONCLUSION: Endorectal MRI can reduce the rate of preoperative understaging from 42% to 22% and it can be used for a given individual because it can detect extraprostatic invasion with 96% specificity, ensuring that very few, if any, patients will be deprived of curative surgery. PMID- 8705220 TI - Infection after transrectal core biopsies of the prostate--risk factors and antibiotic prophylaxis. AB - OBJECTIVE: To compare the infection rate between different durations of antibiotic prophylaxis after transrectal core biopsy and to evaluate the impact of possible risk factors. PATIENTS AND METHODS: The study comprised 491 patients who underwent transrectal core biopsies of the prostate and who were randomized to receive 400 mg of norfloxacin twice daily for one day or one week. RESULTS: Patients receiving prophylaxis for one week had a significantly lower rate of infection (4.9%) compared to patients who received only two tablets (11%; P < 0.05). The most pronounced effect was seen in those patients with risk factors (e.g. an indwelling catheter, a former history of urinary tract infection, diabetes or prostatitis) in whom the infection rate was reduced from 17.9% to 3.3% (P < 0.02), and febrile infections from 9.5% to 1.1% (P < 0.02). CONCLUSIONS: Some factors have a clear impact on the risk of developing an infection after transrectal core biopsy. Prophylaxis for one week with norfloxacin is an effective way to minimize these infections. PMID- 8705221 TI - The impact of pre-donated autologous blood and intra-operative isovolaemic haemodilution on the outcome of transfusion in patients undergoing radical retropubic prostatectomy. AB - OBJECTIVE: To assess the impact of pre-donated autologous blood and intra operative isovolaemic haemodilution on the outcome of transfusions in patients undergoing radical retropubic prostatectomy. PATIENTS AND METHODS: The charts and transfusion records of 192 consecutive patients undergoing radical retropubic prostatectomy were reviewed retrospectively. RESULTS: Of 192 patients, 14% required homologous transfusions, the rate of which varied significantly depending upon blood availability (autologous, haemodilution or homologous) (P < 0.001): 66% of patients with only autologous blood available were transfused a mean of 1.96 units (range 1-6) of blood, of which 87% received autologous alone. 12% both autologous and homologous and 1% homologous blood only. Of the pre donated autologous units, 44% were wasted. CONCLUSIONS: Both the pre-donation of autologous blood and intra-operative isovolaemic haemodilution lower the risk of homologous blood transfusion. It is recommended that patients pre-donate 2 units of autologous blood and undergo intra-operative isovolaemic haemodilution before radical prostatectomy. PMID- 8705222 TI - Orgasm after radical prostatectomy. AB - OBJECTIVE: To evaluate the ability to obtain and the quality of orgasm after radical prostatectomy. PATIENTS AND METHODS: The orgasms experienced after undergoing radical prostatectomy were evaluated in 20 men (median age 65 years, range 56-76) using a semi-structured interview and a self-administered questionnaire. In addition, the patients were asked to write a brief statement about their experiences and sensations during orgasm before and after the operation. RESULTS: Eighteen patients returned the questionnaire and 17 completed a statement indicating what their orgasm was like before and after radical prostatectomy. After the operation, no patient was able to maintain a completely rigid erection, but for five patients the erection was sufficient for sexual intercourse. Nine patients used a vacuum device or intracavernosal self injection. Half the patients reported diminished sexual desire (libido) and arousal after the operation and reported the same to occur in their partners. During their "dry' orgasm post-operatively, none of the patients experienced the exquisite sensation of inevitability, the so-called "point of no return'. Seven of the 14 patients experiencing orgasm complained that their orgasmic sensation was weakened. Four patients reported normal pleasure and sensation compared to that experienced pre-operatively. Surprisingly, nine of the 14 patients had involuntary loss of urine at orgasm; for five of them this was sufficient reason to avoid any sexual contact with their partner. CONCLUSION: Radical prostatectomy may have serious consequences on libido and erectile function but sometimes other important factors, such as the absence of prostate and seminal vesicle contractions, the loss of ejaculation and involuntary loss of urine, may also compromise the orgasm. PMID- 8705224 TI - Treatment of prostate cancer using external beam radiotherapy after laparoscopic pelvic lymph node dissection. AB - OBJECTIVES: To evaluate the results of prostatic irradiation in men with clinically localized prostate cancer and no laparoscopic evidence of nodal metastases compared with a cohort of patients who received radiation therapy with no prior surgical staging. PATIENTS AND METHODS: Thirty-one men with clinically localized prostate cancer and no evidence of pelvic nodal metastases after laparoscopic pelvic lymph node dissection received external beam radiation therapy to the prostate (65-70 Gy). The mean and median prostate specific antigen (PSA) levels in these men before treatment were 41.6 ng/mL and 28.0 ng/mL, respectively, and the mean Gleason sum was 6.1 (range 3-7). During the same interval, a group of 42 consecutive men with clinically localized prostate cancer were treated by external beam radiation therapy with no laparoscopic staging of the pelvic nodes. Treatment failure was defined by the development of bone metastases or a rising PSA level at least 6 months after the completion of radiotherapy. RESULTS: Radiation therapy was generally well tolerated after laparoscopy and no patient required hospitalization or surgery for side-effects related to the treatment. The median duration of follow-up in the 31 men who underwent laparoscopy was 21.5 months. The probability of treatment failure in this group was 41.8% and 56.3% with 24 and 30 months follow-up, respectively. When controlling for pre-treatment PSA level, grade and stage, there was no significant difference in the treatment failure rate between the groups treated with and without laparoscopic staging. CONCLUSIONS: These results suggest that there is no difference in treatment outcome with laparoscopic pelvic lymphadenectomy before external beam radiation therapy in high-risk patients who have significant pre-treatment elevations of PSA level. PMID- 8705223 TI - Bilateral orchidectomy and flutamide versus orchidectomy alone in newly diagnosed patients with metastatic carcinoma of the prostate--an Australian multicentre trial. AB - OBJECTIVES: To investigate the hypothesis that maximal androgen blockade improves the outcome of patients with metastatic prostate cancer. PATIENTS AND METHODS: A total of 222 previously untreated patients with metastatic prostatic cancer were entered into a randomized, double-blind, placebo-controlled trial of bilateral orchidectomy with or without androgen blockade (112 receiving flutamide and 110 a placebo) which commenced in 1985 in four Australian centres. The characteristics of the patients, e.g. age, performance status, the presence of bone pain, duration of disease and the use of prior radiation, were well balanced between the groups. Patients commenced the protocol therapy with flutamide or placebo within the 7 days preceding surgery and continued this medication for a minimum of 2 years, unless there was unequivocal evidence of tumour progression. RESULTS: Apart from a difference in grade 3 or 4 gastrointestinal toxicities between the flutamide and placebo arms (13% and 3%, respectively), serious or life threatening toxicities were uncommon and equally balanced. The assessment of response in six patients (three in each arm) was inevaluable. The objective response rates were 45% and 56% in the flutamide and placebo arms, respectively. There was no difference in survival between the treatments. CONCLUSIONS: This study was not sufficiently powerful to detect small differences in outcome (although the trend in survival favoured the placebo arm) but nevertheless failed to show any benefit for maximal androgen blockade over orchidectomy in this group of patients. PMID- 8705225 TI - Risk factors and mechanism of urethral injury in pelvic fractures. AB - OBJECTIVE: To determine the risk factors and mechanism of urethral injury associated with pelvic fractures. PATIENTS AND METHODS: A total of 203 consecutive male patients with pelvic fracture were studied prospectively, including a clinical examination, radiographic examination of the pelvis, excretory urography and retrograde urethrography. RESULTS: Thirty-nine (19%) patients had urethral injury, five (2.5%) had bladder injury and 12 (6%) had combined urethral and bladder injuries. Urethral injury was by stretching of the intact membranous urethra in 13 patients (25.5%), partial rupture in 13 (25.5%) and complete rupture in 25 (49%). Injury involved the prostatic urethra and bladder neck in three children. Urethral injury was consistently associated with pubic arch fractures. Involvement of the posterior pelvic arch, with fractures of the anterior arch, considerably increased the risk of urethral injury. Also, the risk was greater with an increase in the number of broken rami. CONCLUSION: The highest risk of urethral injury was found in cases with straddle fracture when combined with diastasis of the sacroiliac joint (24 times more than the rest of pelvic fractures); this was followed by straddle fracture alone (3.85 times) and Malgaigne's fracture (3.4 times). Stretching of the membranous urethra usually precedes its rupture, which classically occurs at the bulbomembranous junction. PMID- 8705226 TI - Paroxetine in the treatment of premature ejaculation. AB - OBJECTIVE: To test the efficacy and the adverse effects of a new anti-depressant drug (paroxetine) in the treatment of premature ejaculation. PATIENTS AND METHODS: The study comprised 32 men (mean age 28 years) with premature ejaculation (14 of whom ejaculated before penetration) who were treated with paroxetine (20 mg orally each evening for 2 months). The study group excluded those with neurological and psychiatric disorders, urinary tract infections and drug or alcohol abuse. RESULTS: After about 14 days, the patients' symptoms improved and all patients reported a longer interval before ejaculation. The adverse effects were sleepiness in 19 patients (61%) and mild sensory confusion in 21 (68%), but only one had to withdraw from therapy. Three weeks after the end of therapy, the premature ejaculation recurred in 28 (90%) of the patients. CONCLUSIONS: These results indicate that paroxetine is an effective therapy for premature ejaculation. Further studies with different dosages are necessary to decrease the adverse effects and to prolong the efficacy. PMID- 8705227 TI - A retrospective analysis of 82 cases of cancer of the penis. AB - OBJECTIVE: To identify prognostic factors for penile cancer and to evaluate the treatment strategy for early-stage disease, proposed recently by the European Board of Urology (EBU). PATIENT AND METHODS: The records of 82 patients consecutively referred to the uro-oncological centre at Aarhus University Hospital between 1965 and 1993 were reviewed. The importance of tumour stage, differentiation, patient age, local control and regional lymph node control were assessed using univariate and multivariate analyses. RESULTS: Cox multivariate analysis identified differentiation (odds ratio [OR] = 6.04), UJCC-1978 T-stage (OR = 1.88) and age (OR = 1.04) as independent prognostic variables for survival. Penile amputation in tumours < 4 cm in diameter improved local control but not survival. Regional control and survival were not significantly improved by prophylactic adenectomy. CONCLUSION: Differentiation, T-stage and age were prognostic factors for survival. The results support the EBU treatment strategy involving penis-conserving therapy and watchful waiting for early-stage disease. PMID- 8705228 TI - The role of the Sertoli cell in active immunosuppression in the human testis. AB - OBJECTIVE: To define the involvement of Sertoli cells in immune suppression within the testis by identifying surface receptor molecules that describe or communicate with cells of the immune system. MATERIALS AND METHODS: Ten pairs of human testes obtained from orchidectomy were stained immunohistochemically with established monoclonal antibodies that identify common cells of the immune system. RESULTS: Patterns of staining were similar in all testes. Class I major histocompatibility complex (MHC) antigen, important for self/non-self recognition, was found on Sertoli cells. Class II MHC antigen, important for immune cell interaction and limited to immune cells, was absent. Leu M3, characteristically found on macrophages, was also seen on Sertoli cells but no receptor antigens defining basic T cell types (CD4, CD8), B cells (Leu 12) or natural killer cells (Leu 11) were seen on Sertoli cells. CONCLUSIONS: The Sertoli cell membrane lacks lymphocyte-cell surface markers but harbours somatic cell and macrophage markers. This suggests that immunosuppression within the testis is not maintained through classic lymphocyte receptors. The presence of a macrophage marker is consistent with the known phagocytic activity of the Sertoli cell. PMID- 8705229 TI - Continence after bladder-neck reconstruction in patients with bladder exstrophy and pubic diastasis. AB - OBJECTIVE: To review the results of bladder-neck reconstruction in patients with repaired bladder exstrophy and pubic diastasis. PATIENTS AND METHODS: Nine girls (mean age 7 years, range 4-17) and four boys (mean age 9 years, range 5-15) underwent a modified Young-Dees-Leadbetter bladder-neck reconstruction with augmentation cystoplasty (YDL-C). The patients were reviewed retrospectively (follow-up, 1-6 years) to assess continence, particularly in relation to the degree of pubic diastasis measured on an appropriate abdominal radiograph. RESULTS: Ten patients were continent; seven girls and one boy are managed by clean intermittent catheterization (CIC) and one girl and one boy void normally. One girl who would not allow CIC and one boy in whom CIC was not possible are incontinent and are scheduled for construction of a continent diversion. One incontinent boy who also would not allow CIC was lost to follow-up. Public diastasis had no bearing on the surgical results, the 10 continent patients having diastasis ranging from 4 to 9 cm (mean 5.5 cm) and the incontinent patients a diastasis of 3.4 and 6.5 cm (mean 4.5 cm). CONCLUSION: Young-Dees Leadbetter bladder-neck reconstruction with augmentation cystoplasty is a satisfactory operation in patients with bladder exstrophy. We believe that the rate of continence reflected a competent tubularization with an adequate bladder capacity and assured bladder emptying. A closed pelvis with approximated public bones is not necessary to achieve this objective. PMID- 8705230 TI - The frequency of posterior urethral valves in Oman. AB - OBJECTIVE: To ascertain the frequency of posterior urethral valves (PUV) in Oman, using hospital records. MATERIALS AND METHODS: The resident population of Oman is 1.5 million and all complex paediatric surgical cases are referred to a single unit in the capital city of Muscat. A register of patients has been kept in that unit since 1988. Hospital case-records were reviewed to determine associated abnormalities, date and place of birth, tribal origin and, from further interviews, the family pedigree and degree of consanguinity. RESULTS: This retrospective survey identified 56 Omani children born between 1989 and 1994 who were treated in the Paediatric Surgery Department at the Royal Hospital, Muscat for congenital PUV. The calculated incidence was 1 in 2375 newborn males, which is 2-4 times higher than that previously reported in other populations. There was no relation to the estimated month of conception and therefore environmental hyperthermia due to the extreme ambient temperature does not seems not to be a causative factor. There was consanguinity in 40 of 46 cases (87%) where a complete family history was available. Parents were first cousins in 54% of cases, compared with the background rate of 24% in over 6000 families interviewed in the antenatal clinics at six hospitals in Oman. There was also a concentration of PUV within specific tribes. About one-third of the cases also had associated congenital abnormalities, the most frequent of which were genitourinary (17%), but gastrointestinal (9%) and spinal (7%) defects were also observed. CONCLUSION: The frequency of PUV in newborn Omani males was 1 in 2375, which is considerably higher than any previously reported series. This was associated with an increased rate of consanguinity, but there was no clear pattern of inheritance. PMID- 8705231 TI - Failure of pre-natal ultrasonography to prevent urinary infection associated with underlying urological abnormalities. AB - OBJECTIVES: To analyse the reasons underlying the failure of routine pre-natal ultrasonography to prevent the subsequent development of urinary tract infection (UTI) in children with predisposing urological abnormalities. PATIENTS AND METHODS: This retrospective study comprised 39 children (22 females and 17 males) who had at least one documented UTI, the presence of an anatomical anomaly of the urinary tract recognized as predisposing to UTI and had undergone ultrasonography of the urinary tract undertaken in fetal life as part of routine maternal ante natal ultrasonography. Four categories of patients were defined: Group A, those with normal findings on pre-natal ultrasonography and no urological abnormality detected; Group B, those with a urological abnormality detected but where there was a subsequent failure of communication among clinicians; Group C, those with a urological abnormality but who received inappropriate or sub-optimal post-natal management and; Group D, those with a urological abnormality but who had a UTI despite appropriate post-natal management. In each case, the most severe documented episode of UTI was categorized as: Grade I, asymptomatic bacteriuria; Grade II, mild/moderate symptomatic UTI and; Grade III, severe symptomatic UTI necessitating hospital admission. RESULTS: Group A comprised 22 (56%), Group B three (9%), Group C two (5%) and Group D 12 children (31%). Of the 22 children in Group A, nine experienced a UTI of sufficient severity to necessitate hospital admission. Of the 12 children in Group D only one required hospital admission. CONCLUSIONS: The failure of pre-natal ultrasonography to identify the underlying predisposing urological abnormality was the most important factor contributing to subsequent UTI in post-natal life. Failure of communication and inappropriate post-natal management were numerically unimportant. In some children, UTI occurred despite pre-natal detection of their underlying anomaly and appropriate post-natal management. However, in this group the UTI was less severe than in those children whose urological anomalies had not been detected by pre-natal ultrasonography. PMID- 8705233 TI - Epidural needle and catheter for bladder instillation of a marker dye. PMID- 8705232 TI - Laparoscopic dismembered pyeloplasty in children: preliminary results. AB - OBJECTIVES: To investigate the technique of laparoscopic pyeloplasty in children with pelvi-ureteric junction (PUJ) obstruction. PATIENTS AND METHODS: Six children, aged between 2.5 and 15 years, were treated by laparoscopic pyeloplasty in a prospective study. The outcome was assessed by post-operative isotopic renography. Four of the children underwent a Y/V plasty of the PUJ structure and one a Hynes-Anderson dismembered pyeloplasty. RESULTS: One case (the first in this series) was converted to open pyeloplasty due to adhesions around the PUJ from previous surgery. One patient developed an abdominal haematoma at the trocar site, which resolved without treatment. All anastomoses were completed without complication. Five of the children had normal or significantly improved drainage times on post-operative renography. In one patient, the drainage time deteriorated post-operatively, although differential function of the operated kidney improved. CONCLUSIONS: These early results suggest that laparoscopic pyeloplasty in children is feasible, safe and effective; a longer term follow-up is awaited. PMID- 8705234 TI - Push-ups: a new technique in intravenous urography. PMID- 8705235 TI - A case of Cowden's disease accompanied by prostatic cancer. PMID- 8705236 TI - Haemoperitoneum due to necrosis of bulky retroperitoneal metastases: an unusual complication of chemotherapy for testicular cancer. PMID- 8705237 TI - High-performance aircraft--a possible cause of male infertility. PMID- 8705238 TI - Intracavernosal administration of papaverine in a blind patient. PMID- 8705239 TI - Inguinal lymph node metastases from a testicular tumour. PMID- 8705240 TI - Amputation of distal penile glans during neonatal ritual circumcision--a rare complication. PMID- 8705241 TI - Extensive bladder infarction in a strangulated inguinal hernia. PMID- 8705242 TI - Renal angiomyolipoma with extension into renal vein. PMID- 8705243 TI - Posterior urethral valves in adults diagnosed by ultrasonography. PMID- 8705244 TI - Congenital vesicocutaneous fistula following antenatal bladder aspiration. PMID- 8705245 TI - Shared care for benign prostatic hyperplasia: a feasibility study. PMID- 8705246 TI - Neurogenic bladder induced by brain abscess. PMID- 8705247 TI - Nanoparticulate systems in drug delivery and targeting. PMID- 8705248 TI - Mucosal penetration enhancement. PMID- 8705249 TI - The application of cell culture systems in drug discovery and development. PMID- 8705250 TI - Bioadhesives for optimization of drug delivery. PMID- 8705251 TI - Delivery of oligonucleotides and polynucleotides. PMID- 8705252 TI - The lack of effect of induced net fluid absorption on the in vivo permeability of terbutaline in the human jejunum. AB - The absorption mechanism(s) of terbutaline in the human jejunum was studied by using the intestinal perfusion instrument, Loc-I-Gut. The present study was divided into three parts. In Part I the absorption of 10 and 20 microM of both (+) and (-)-terbutaline enantiomers was studied. The influence of D-glucose (80 mM) on the net fluid transport across the intestinal wall and the effective intestinal permeability (Peff) of both (+/-)-terbutaline (10 microM) and antipyrine (0.5 mM) was investigated in Part II. The experimental design of Part III was similar to that in Part II, with the exception that the perfusion solution was hypotonic and had a D-glucose concentration of 80 mM. No statistical differences were found in the Peff between terbutaline enantiomers or their concentrations. D-glucose (80 mM) did neither affect net fluid transport nor the Peff of (+/-)-terbutaline and antipyrine in the human jejunum. In contrast, hypotonic D-glucose (80 mM) solution induced a net fluid absorption (p < 0.01). In parallel with this observation, the Peff -value of (+/-)-terbutaline was unchanged, whereas the absorption of antipyrine was found to be significantly increased (p < 0.05). The increased permeability of antipyrine during the net fluid absorption phase might be due to convective paracellular flow, but more likely is it a consequence of a higher concentration gradient of the drug close to the intestinal wall, and thereby increased transcellular absorption. Based on these findings we propose that the major route for the oral absorption of terbutaline and antipyrine might be passive transcellular diffusion. PMID- 8705253 TI - Prevention of phorbol myristate acetate-induced acute lung injury by alpha tocopherol liposomes. AB - Phorbol-myristate acetate (PMA) is commonly used to produce experimental edema and other tissue injuries in the lung. Lung injuries induced by the administration of PMA has been shown to be mediated mainly by neutrophils. Neutrophils recruited to the lower respiratory tract may damage lung tissues by releasing reactive oxygen species, neutral proteases, and lysosomal enzymes. The present study was conducted to investigate whether alpha-tocopherol, entrapped in dipalmitoylphosphatidylcholine liposomes and delivered directly to the lung, could counteract some of the PMA-induced lung injuries. Plain liposomes or alpha tocopherol containing liposomes (8 mg alpha-tocopherol/kg body weight) were intratracheally instilled into the lungs of rats 24 hr prior to PMA exposure (25 micrograms/kg) and treated rats were killed 3 hr later. Lungs of control animals exposed to PMA developed an increase in lung weight and lipid peroxidation as well as a decrease in lung angiotensin converting enzyme (ACE) and alkaline phosphatase (AKP) activities. PMA treatment also caused an increase in myeloperoxidase (MPO) activity in the lung, suggestive of neutrophil infiltration. Pretreatment of PMA-treated rats with plain liposomes had no effect on PMA-induced injuries. In contrast, pretreatment of rats with liposomal alpha tocopherol, 24 hr prior to PMA administration, resulted in a significant elevation of pulmonary alpha-tocopherol concentration, accompanied by a concomitant reduction in MPO activity and reversal of PMA-induced changes in lung edema, lipid peroxidation, ACE and AKP activities. These results appear to demonstrate that the intratracheal administration of a liposome-associated lipophilic antioxidant, such as alpha-tocopherol, can significantly ameliorate the toxic effects of reactive oxygen species, putatively released from PMA stimulated pulmonary target cells and infiltrating neutrophils. PMID- 8705254 TI - The delivery of insulin from aqueous and non-aqueous reservoirs governed by a glucose sensitive gel membrane. AB - A self regulating delivery device, responsive to glucose, has been shown to operate successfully in vitro. This comprises a gel membrane which determines the delivery rate of insulin from a reservoir. The gel consists of a synthetic polysucrose and the lectin, concanavalin A. The mechanism is one of displacement of the branched polysaccharide from the lectin receptors by incoming glucose. The gel loses its high viscosity as a result but reforms on removal of glucose, thus providing the switch controlling the drug diffusion rate. The drug does not require to be chemically modified and thus the device is adaptable to other anti hyperglycaemic drugs. However, results here indicate that the molecular weight of the solute may be an important parameter. Others include path length, gel formulation and temperature. It had been hypothesised that the reversal might be improved by the use of a non-aqueous reservoir of insulin. However, with the use of insulin, the switching off was found to be superior to that found with other test solutes used in previous studies, irrespective of the reservoir solvent. The advantages in the use of the non-aqueous system include, however, more reproducibility in the magnitude of response and a reduced temperature sensitivity. PMID- 8705255 TI - Translocation of particulates across the gut wall--a quantitative approach. AB - The translocation of latex particles across the epithelium of the rat small intestine and their transfer to internal organs was determined and quantified. The rate of particle uptake was also established. Particle uptake from the gut lumen to internal tissues was rapid as they were detected in all tissues examined within 30 minutes of oral administration. The maximum number of particles per gram tissue was detected in tissues of the small intestine. Particle number in spleen, kidney, lung, liver and brain increased with time, and in mesenteric lymph node, and heart tissues they decreased with time. Particles were also detected in bone marrow samples. These findings suggest that this animal model is useful in the evaluation and quantification of particle uptake and the determination of the tissues to where they are transported. PMID- 8705256 TI - O/W lipid emulsions for parenteral drug delivery. II. Effect of composition on pharmacokinetics of incorporated drug. AB - The potential usefulness of O/W lipid emulsions as injectable drug delivery systems for lipophilic drugs was examined using a model lipophilic drug, sudan II (clogP = 5.4) in the normal rats. The standard lipid emulsion composed of soybean oil and egg yolk phosphatides increased the blood concentration of sudan II after i.v. injection when compared with its solubilized solution by plasma. However, it was still lower than that of the oil particles, and the distribution of sudan II to liver, lungs, adipose tissue, heart, and muscle was not altered, and only that to brain and kidneys was decreased. Herein, the effect of extensive alterations in the lipid emulsion composition on the blood concentration and organ distribution of sudan II was examined in comparison with the standard formulation. Addition of cholesterol, use of pure egg yolk phosphatidylcholine, use of phospholipids with saturated alkyl chain, use of saturated long chain triglycerides, and use of saturated medium chain triglycerides were tested. The oil particles of all tested lipid emulsions were still located in plasma space, and use of saturated medium chain triglycerides was the most effective way to increase blood concentration of sudan II, resulting in higher distribution to liver, lungs, spleen, and brain. This was caused by the increase of the steady state partition of sudan II to the oil particles, and not by alteration of their organ distribution clearance. PMID- 8705257 TI - Pharmacokinetics and pharmacodynamics of recombinant human granulocyte colony stimulating factor (rhG-CSF) following intranasal administration in rabbits. AB - The objective of this study was to evaluate the pharmacokinetics and pharmacodynamics of G-CSF as well as their relationship following intranasal (i.n.) administration of aqueous rhG-CSF preparations with or without additives. In order to achieve a better understanding of the dosage regimen and the effectiveness of intranasally administered rhG-CSF in inducing leukopoiesis, we investigated rhG-CSF absorption and blood leukocyte dynamics with respect to dose in rabbits. RhG-CSF could be absorbed through the nasal cavity of rabbits when rhG-CSF aqueous preparations, especially those containing alpha-cyclodextrin (alpha-CyD) were intranasally administered. We found that serum G-CSF levels and the total count of leukocytes in peripheral blood (total blood leukocyte count) showed a dose-dependent increase with rhG-CSF. The area under the serum G-CSF concentration-time curve (AUC, a pharmacokinetic parameter) and the area under the increased total blood leukocyte count-time curve (AUL, a pharmacodynamic parameter) increased with increase of dose of rhG-CSF administered intranasally. Good agreement was observed between log AUC and AUL; thus, it is concluded that an increase of AUC leads to an increase in effectiveness of rhG-CSF in inducing leukopoiesis in rabbits. A novel rhG-CSF delivery system in the form of i.n. administration of rhG-CSF was thus achieved. PMID- 8705258 TI - Gastrointestinal adenomatous polyps. AB - Adenomatous polyps of the gastrointestinal tract are dysplastic precursor lesions of adenocarcinoma. The features of an adenoma that are associated with a greater tendency to progress to carcinoma include larger polyp size, high grade dysplasia, and increased villous glandular architecture. Alterations in particular oncogenes and tumor-suppressor genes have been correlated with various stages of colonic adenoma formation, thereby facilitating our knowledge of adenoma pathogenesis. Heredity and environment contribute to the risk of developing adenomatous polyps of the colon. The incidence of colorectal cancer can be decreased by the endoscopic removal of adenomas, thereby providing good rationale for screening and surveillance programs. PMID- 8705259 TI - Barrett's esophagus. AB - When the distal esophagus is lined by a segment of columnar epithelium that is abnormal either because it is extensive or because it has intestinal features, the condition is called Barrett's esophagus. Traditionally, Barrett's esophagus has been associated with severe gastroesophageal reflux disease (GERD) and with a high risk for adenocarcinoma. Recently, investigators have shown that short segments of intestinal metaplasia can be found frequently in the distal esophagus of patients who have few signs and symptoms of GERD. For these patients, the risk for carcinogenesis is not clear. This article summarizes the diagnostic and clinical features of Barrett's esophagus and presents an approach to the management of patients with this fascinating disorder. PMID- 8705260 TI - Premalignant lesions of the stomach. AB - Recent studies indicate that chronic gastritis induced by Helicobacter pylori infection is a predisposing condition for the development of both gastric adenocarcinoma and primary gastric B-cell mucosal associated lymphoid tissue (MALT) lymphoma. The chronic gastritis induced by H pylori may evolved in specific individuals in response to additional genetic, environmental, and bacterial-virulence factors. The transitions from chronic gastritis to gastric atrophy with small intestinal (type I) metaplasia, incomplete small intestinal (type II) metaplasia, large intestinal (type III) metaplasia, low-grade dysplasia, and finally high-grade dysplasia are associated with respectively increasing cancer risk. H pylori infection may also lead to the development of an abnormal clone of B-cells and formation of low-grade MALT lymphoma, which is thought to be a precursor for the development of metastatic high-grade MALT lymphoma. In some patients, low grade MALT lymphoma has been shown to regress after eradication of the H pylori infection. The specific genetic abnormalities associated with these premalignant stages of gastric carcinogenesis are currently being defined. Other conditions associated with an increased risk of gastric adenocarcinoma include autoimmune atrophic gastritis, adenomatous polyps, postgastrectomy remnants, and possibly Menetrier's disease. PMID- 8705261 TI - Preneoplastic conditions of the liver. AB - Hepatocellular carcinoma (HCC) is one of the most common cancers in the world. The majority of patients who develop HCC have underlying cirrhosis, which suggests that cirrhosis itself represents a preneoplastic condition. Nevertheless, whereas patients with cirrhosis of any origin are at increased risk of developing HCC, those with chronic hepatitis B or C infection seem to be at greatest risk. Patients with cirrhosis resulting from chronic alcohol use, hemochromatosis, autoimmune hepatitis, or alpha-1 antitrypsin deficiency have less risk of developing this cancer, and some hepatic diseases, such as primary biliary cirrhosis and Wilson's disease, do not predispose affected persons to an appreciable risk of developing HCC. Certain histological features, such as liver cell dysplasia and macroregenerative nodules, may represent preneoplastic alterations of hepatocytes, but these changes do not seem to be a necessary step in the evolution of liver cancer. The pathogenesis of HCC is unclear, but seems to involve several steps. Hepatitis B virus infection may result in the malignant transformation of hepatocytes by some directly oncogenic mechanism, whereas other necroinflammatory conditions probably predispose to the development of HCC through the introduction of genetic alterations coupled with a reduction of genetic repair functions. Screening patients at risk for the development of HCC using alpha fetoprotein measurements and ultrasonography is widely practiced despite inconclusive evidence of efficacy. If screening is performed, the program used should be tailored to the perceived risk for a particular patient. PMID- 8705262 TI - Premalignant conditions of the small intestine. AB - Cancer of the small intestine is rare compared with other sites in the gastrointestinal tract. Of the four major primary small-bowel tumors (adenocarcinomas, lymphomas, carcinoid, and leiomyosarcomas), adenocarcinomas and lymphomas are associated with diseases that seem to increase the risk of developing these malignancies. In the case of immunoproliferative small intestinal disease and celiac disease, both of which are thought to predispose patients to the development of primary lymphoma, treatment of the predisposing conditions seems to decrease the risk of developing subsequent malignancy. Recognition of the increased risk associated with other conditions, such as immunodeficiency syndromes, nodular lymphoid hyperplasia, Crohn's disease, the gastrointestinal polyposis syndromes, hereditary nonpolyposis colon cancer, neurofibromatosis, long-standing ileostomy, and urinary diversion procedures, may lead to early diagnosis and improved survival. PMID- 8705263 TI - Inflammatory bowel disease. AB - Patients with extensive ulcerative and Crohn's colitis represent a group at high risk for developing colorectal cancer. Two clear independent risk factors for developing colorectal cancer in patients with chronic colitis include duration and extent of disease. Cancers in chronic colitis do not develop from a colonoscopically recognizable adenomatous polyp similar to that in sporadic colon cancer, but instead arises from flat dysplastic epithelium that is typically not colonoscopically distinguishable from adjacent nondysplastic epithelium. All patients with extensive disease require active management of their increased cancer risk. Active management of cancer risk in chronic long-term colitis should not be presumed to be equivalent to colonoscopic surveillance. There is little data with which to reassure a patient regarding the efficacy of colonoscopic surveillance. A reasonable alternative to colonoscopic surveillance in patients with ulcerative colitis is the restorative proctocolectomy (ileoanal pull through), which maintains continence and avoids a stoma and appliance. If colonoscopic surveillance is undertaken, a clear understanding of what the definition of a positive surveillance test, ie, when surgical action is taken, is pivotal to the success of the surveillance program. There are now ample data confirming that the finding of any unequivocal dysplasia (low- or high-grade) is associated with a high risk of coexistent or future colorectal cancer. Dysplasia confirmed by a second pathologist (preferably an experienced gastrointestinal pathologist) should prompt a recommendation for colectomy. Future cancer surveillance in chronic colitis will almost certainly involve some applied molecular genetic test. PMID- 8705264 TI - Antibiotic associated nephropathy. AB - Renal biopsies of 43 patients who developed renal complications after treatment with antibiotics were studied. The treatment with antibiotics in these cases was used for many different reasons such as: bronchitis, bronchopneumonia, cystitis, tonsillitis, sepsis, peritonitis, gangrene of the foot and tuberculosis. The renal function of these patients, before the treatment with antibiotics was normal. The biopsies were studied by light, electron and immunofluorescence microscopy. In 43 cases treated with antibiotics renal changes were shown. Three types of morphologic changes were found: acute tubular necrosis (ATN) (13 cases), acute tubulo-interstitial diseases (ATID) (21 cases), focal glomerulonephritis with crescents (FGN) (9 cases). The renal pathologic changes were most commonly seen in patients treated with 2 groups of antibiotics: aminoglycosides (21 cases) and antibiotics of the penicillin group (15 cases). The most characteristic feature of aminoglycosides is their direct toxic effect leading to ATN. Antibiotics of the penicillin type more commonly caused an allergic reaction leading to ATID (secondary to cellular mechanisms) or FGN (secondary to a predominantly humoral mechanism). Renal changes in the use of other antibiotics were much less manifest and were usually due to a hypersensitivity reaction. Cephalosporins, if used in combination with other antibiotics can increase their nephrotoxicity. PMID- 8705265 TI - The histological appearance of the periprosthetic capsule in failed total hip arthroplasty differs depending on the presence of polyethylene acetabulum destruction, iliac bone damage and presence of infection. AB - Aseptic loosening of the prosthesis is the most common complication of the total hip replacement. The tissues resected on revision surgery often contain polyethylene debris, originating from the worn prosthesis, as well as the fragments of acrylic cement. These foreign bodies are accompanied by various patterns of inflammatory reaction. We compared the elements of the histological picture of periprosthetic capsules from 20 patients with the clinical data, the appearance of the failed prosthesis and the state of the surrounding bone. As a result, we were able to identify characteristic morphological features linked with the aseptic implant loosening compared to the infection group. Among the aseptic implant loosening cases, those with the polyethylene acetabulum damage were associated with the more florid giant cell reaction and diffuse hyalinization of granulation tissue. The iliac bone damage was usually accompanied by more abundant cement fragments, lymphoid infiltrate and solid acellular islands of hyaline. Thus we suspect, that there are at least two mechanisms leading to aseptic failure of the total hip replacement. PMID- 8705266 TI - Pathogenesis and morphogenesis of microcirculatory disorders in myocardial infarction. Clinical and ultrastructural examination. AB - Major advances in the understanding of myocardial infarction (MI) have been made on animal models of ischemia and infarction. We performed ultrastructural examinations of the hearts of 36 patients who died during the acute phase of MI. Tissue for investigation was obtained by express necropsy immediately after death at the clinic. All components of the microcirculatory system of the heart were damaged during the acute phase of MI. Coronary thrombosis led to arteriolar spasm and intravascular blood coagulation in the vicinity of the damaged artery. Microvessels reduction in ischemic, necrotic, and near-infarct zones was caused by endothelial cell injury and rheological disturbances. Thrombocytes played a decisive role in thrombus formation and arteriolar constriction. Leukocytes nearly always impaired the microhemodynamics in ischemic zones due to their size and rigidity. Microcirculatory disorders seemed to precede myocardial cell injury and death. PMID- 8705267 TI - p53 expression in sweat gland tumors. AB - We analyzed the expression of p53 in 74 cutaneous adnexal tumors, with enhancement of the detection by incubation of the slides in the microwave. The immunostaining in benign tumors was almost uniformly negative as we found p53 positivity only in one poroma, one nodular hidradenoma, and one case of syringocystadenoma papilliferum (amongst 13 spiradenomas, 9 cylindromas, 12 nodular hidradenomas, 7 poromas, 6 syringomas, 7 syringocystadenomas papilliferum, 2 papillary tubular adenomas and 4 chondroid syringomas). These results contrasted with the widespread p53 overexpression, which was revealed in the sweat gland carcinomas. All spiradenocarcinomas (3), malignant nodular hidradenoma (1), apocrine hidradenocarcinoma (1), and malignant syringoadenoma (1) showed a strong reaction to anti-p53 antibody. Two of three eccrine hidradenocarcinomas, and two of three porocarcinomas presented p53 overexpression, whereas in one case of malignant cylindroma and adenoid cystic carcinoma we did not find p53-positivity. The results of the study indicate an important role, that p53 protein plays in the malignant sweat gland tumors in comparison to their benign counterparts, but reveal that its overexpression may also occur in the reactive and benign neoplastic processes. PMID- 8705268 TI - Hyalinizing clear cell carcinoma of salivary gland. A case report. AB - Authors describe a case of hyalinizing clear cell carcinoma of the parotid gland in a 65-year-old female. Morphological characteristics of clear cell tumors of salivary glands are then discussed. PMID- 8705269 TI - The immunohistochemistry and ultrastructure of ganglioglioma with chromosomal alterations: a case report. AB - Ganglioglioma, together with its counterparts-ganglioneuroma and gangliocytoma are relatively uncommon neoplasms of the brain composed of neoplastic neurons (ganglion and ganglioid cells) and glial cells. We report here a case of ganglioglioma studied by electron microscopy. The case was further characterized by peculiar chromosomal alterations, 46,XX[6]/43,XX[1], der(1)t(1;5)(q21;q12), der(8;13)(q10;q10),-9,i(10)(q10). Routine light microscopy revealed mixed neuro glial tumor composed of pilocytic astrocytes with abundant Rosenthal fibers and relatively numerous ganglion cells. The latter were immunoreactive with antibodies (Abs) against synaptophysin and neurofilament protein (NFP). Anti-NFP Abs also immunostained numerous distorted axons embedded in the tumor mass. Some of these showed bullous swellings and thus were analogous to dystrophic neurites or spheroids. Ganglion cells were characterized by abundant intracytoplasmic dense-core vesicles, absence of intermediate filaments and numerous microtubules. Occasionally a close apposition of ganglion cells and Rosenthal fibers were seen. Dense-cored vesicles were pleomorphic and ranged in diameter from small synaptic vesicles to large lysosome-like neurosecretory granules. The former occasionally formed characteristic dumbbell shapes. Neoplastic astrocytes were identical to those of other glial tumors of astrocytic lineage; numerous Rosenthal fibers were frequently seen. PMID- 8705270 TI - Myxoid liposarcoma of the urinary bladder. AB - We report a case of myxoid liposarcoma of the urinary bladder, that occurred in a 36-year-old woman. We faced the difficulties in reaching the preoperative diagnosis, likewise in determining the extent of the tumor, as the results of the computed tomography differed from the pathologic findings of the tumor. The review of the literature concerning this unusual site for liposarcoma and methods of treatment is also included. PMID- 8705271 TI - Diagnostic significance of the myoepithelial cells in the benign and malignant sweat gland tumors. AB - We studied the presence of myoepithelial cells (MCs) in the benign and malignant sweat gland adenomas by immunostaining with antibody to alpha-smooth muscle actin. We found peripheral arrangement of MCs in the neoplastic nests in cutaneous cylindromas (9), spiradenomas (13), syringoadenomas (7), and one case of adenoid cystic carcinoma. Dispersed MCs were identified in some cases of nodular hidradenoma (7/13) and malignant portion of spiradenocarcinoma (2/3). The remnants of the peripheral arrangement of MCs were retained in the benign part in cylindrocarcinoma and every case of spiradenocarcinoma, likewise in composed malignant adnexoma (malignisation within cylindroma/spiradenoma). We could not find MCs in papillary eccrine adenoma (2), poromas (7) and porocarcinomas (3), syringomas (6), chondroid syringomas (3), malignant nodular hidradenoma (1) and malignant portion of syringoadenoma (1). In one of three cases of eccrine hidradenocarcinoma focal myoepithelial differentiation could be identified at the periphery of the epithelial nests. These results confirm heterogeneity of the differentiation in sweat gland tumors, emphasizing the validity of division of those tumors into those with the differentiation towards secretory and ductal portion of the gland. The former group demonstrates the variability of arrangement of myoepithelial cells depending on the degree of dedifferentiation of the developing carcinoma. These cells may be replaced by outgrowing tumor, but may also accompany the neoplastic growth. Less significant is the presence of the MCs in the adenomas with ductal differentiation, which either in benign and malignant tumors may be lacking. The variability of MCs occurrence in cutaneous adenomas and carcinomas precludes its significance as a solitary factor in the differentiation between benign and malignant proliferations. PMID- 8705272 TI - Uptake of phenylalanine by the rat brain after resuscitation from cardiac arrest. AB - In 47 adult rats 10-min cardiac arrest was induced by the intrathoracic compression of the heart vessel bundle. The animals were sacrificed at 15, 30, 60, 120 min and 6 h or 1, 3, 7 days after resuscitation. Decapitation was performed 15 sec after intracarotid injection of mixture of L-[U-14C] phenylalanine (PHE) and tritiated water in PBS buffer. By the dual label method the brain uptake index (BUI) and percent of injected dose of amino acid in the cerebral hemisphere were calculated. A decrease of PHE uptake and drop of BUI revealed the blood-brain barrier (BBB) alterations resulting in diminution of amino acid transport into brain. The most pronounced changes developed between 15 and 120 min after resuscitation and also after 7 days. The above data revealed the decreased active transport of PHE in the early and late periods after ischemic insult. PMID- 8705273 TI - Reactive and degenerative changes of tissues surrounding a brain tumor. AB - We report here immunohistochemical and ultrastructural studies of the pattern of brain degeneration being a consequence of the presence of brain tumors. Robust microglial reaction with upregulation of MHC II type antigens within and around the brain tumor were seen along with the purely degenerative phenomena like neuroaxonal dystrophy (NAD) and myelin dilatation ("ballooning"). The reaction was monotonous and independent of the histological type of the brain tumor. PMID- 8705274 TI - Apoptosis, programmed cell death in the central nervous system. AB - Cell death by apoptosis occurs in a number of normal tissue systems including the nervous system. It is a highly specific process, which requires new gene expressions and the production of new proteins. The genetic information determines biochemical and morphological sequence of events in the cell undergoing apoptosis. This review focuses on the recent informations concerning the participation of the genes in the survival and death of neurons in the central nervous system and the effect of the anti-cancer drugs on the induction of apoptosis in the cell. PMID- 8705275 TI - Morphological study of microglia in human mesencephalon during the development and aging. AB - To assess the cytogenesis and the structure of the microglial cells, we studied mesencephalons in 47 human fetuses at 7th-40th week of gestational age, and in 18 adult brains from 20 to 70 years. The microglial cells were identified and characterized by morphological criteria using immunohistochemical and histochemical techniques. As early as in the 8th week of gestational age RCA-1 positive cells were detected, mainly in form of amoeboid microglial cells. These microglial cells were observed around the germinal matrix, and at or near the wall of blood vessels. RCA-1 positive cells which were detected within leptomeninges were large but without processes. At the 16th-40th week of gestational age we observed in mesencephalon, amoeboid microglial cells, and also RCA-1 positive and ferritin reactive ramified microglial cells. PMID- 8705276 TI - Creutzfeldt-Jakob disease: ultrastructural study of brain biopsy: unusual interaction between astrocytes and oligo- and microglia. AB - We report a case of Creutzfeldt-Jakob (CJD) disease in a 72-year-old man without characteristic EEG alterations. Unusually abundant pleomorphic PrP-immunopositive amyloid plaques were found in the cerebellum. Despite of this similarity with Gerstmann-Straussler-Scheinker (GSS) disease the case cannot be classified as GSS lacking classical GSS 102 PRNP gene mutation. The ultrastructural findings were similar to those described previously for CJD. The close contact of reactive astrocytes with both oligodendroglia and microglia in this case suggests their possible functional interaction. PMID- 8705277 TI - A case of progressive supranuclear palsy with widespread appearance of neurofibrillary changes and associated senile and vascular brain lesions. AB - A case of progressive supranuclear palsy in association with vascular and senile brain changes in a 70-year-old woman is described. Neuropathological study with immunocytochemistry anti-tau-1 revealed widely distributed neurofibrillary tangles (NFT) and neuropil threads (NT) in several subcortical nuclei, including pallidum, subthalamic nucleus, substantia nigra, brain stem tegmentum and, to a lesser extent, in cerebral cortex. Moreover, the tau-1 positive NT were observed in many fiber bundles of the subcortical white matter. All NFT and NT were immunonegative against ubiquitin. Electron microscopic study disclosed straight filaments of about 15 nm diameter in the axoplasm of large myelinated fibers. Ultrastructural findings and appearance of abnormal tau in the white matter indicate an extension of characteristic cytoskeletal pathology with subcortical projection fibers involvement in the presented case. PMID- 8705278 TI - Neural cell adhesion molecule (N-CAM) as a marker of muscle tissue alternations. Review of the literature and own observations. AB - In this paper the expression of neural cell adhesion molecule (N-CAM) analyzed by indirect immunofluorescence on human skeletal muscle is reported. In adult human muscle N-CAM is concentrated near neuromuscular junctions and on satellite cells but is nearly undetectable in non synaptic portions of myofibers. N-CAM is abundant on myotubes and intramuscular nerves in embryonic muscle but is lost as development proceeds. After denervation N-CAM appears along the entire length of muscle fibers. In addition N-CAM is observed on the surface of regenerating muscle cells. These data suggest that N-CAM immunohistochemistry may play complement conventional technique in the diagnosis of neuromuscular diseases. PMID- 8705279 TI - Overexpression of amyloid precursor protein of Alzheimer's type in brains of intoxicated children. AB - The coincidence of neuronal stress induced by intoxication and an overexpression of amyloid precursor protein (APP) in the brains of children was examined. Brains of ten children accidentally intoxicated by poisonous mushroom were studied by means of immunohistochemical methods using monoclonal antibodies generated against different domains of APP and glial cell markers. Overexpression of APP was found in the brain neurons of all intoxicated children. Neurons were immunopositive with the antibodies generated against the middle (amyloid beta protein) domain of APP. No extracellular deposits were found in the tissue. Our results provided, for the first time, the evidence that overexpression of APP concomitant with the neuronal stress is age-independent phenomenon appearing not only in the brain of adults but in very young individuals as well. PMID- 8705280 TI - Sarcomatosis of leptomeninges, brain and spinal cord coexisting with von Recklinghausen's neurofibromatosis. A case report and review of the literature. AB - A case of 56-year-old male with sarcomatosis of leptomeninges as well as of the brain and spinal cord coexisting with Recklinghausen's neurofibromatosis is presented. Neurological and neurophysiological symptoms of the disease resembled those of amyotrophic lateral sclerosis (ALS). Patient died 7 months after onset of the initial symptoms. The post-mortem examination revealed neoplastic infiltration of the leptomeninges of brain and spinal cord. Histologically sarcomatosis of the leptomeninges was diagnosed and immunohistochemical analysis of the neoplastic infiltrates can indicate fibrohistiocytic origin of the neoplasm, suggesting also a probable contribution of perineurial cells in the pathogenesis of the tumor. On the grounds of the performed immunohistochemical study together with a review of the literature, the differential diagnosis of malignant mesenchymal tumors of the CNS is discussed with a special regard to their histogenesis. PMID- 8705281 TI - Neurofibromatosis type 2. Case report. AB - A case of a 24-year-old woman with peripheral paresis of the facial nerve, balance disturbance, hearing loss and epileptic seizures for many years is presented. At time of admission to hospital cerebral magnetic resonance imaging (MRI) showed several large tumors situated supra- and infratentorially. Histological examination of the operated tumors revealed bilateral acoustic schwannomas in the cerebellopontine angles and mixed meningioma in the others. The patient was diagnosed as neurofibromatosis type 2 (NF2) according to clinical criteria for neurofibromatoses. Several months after the last operation, she exhibited weakness of all extremities. On spinal MRI an intramedullary tumor in the cervical region and additional focal lesions along central canal were found. Surgical therapy was not performed because of clinical improvement after dexamethasone treatment and location of lesion in cervical medulla. Our case confirms frequently occurring lack of neurocutaneous changes and late appearance of significant neurological symptoms in NF2. PMID- 8705282 TI - DNA protein flow cytometry of dissociated cultures of human anaplastic gliomas. Pattern of proliferation and differentiation, and the effect of a new cytostatic drug cladribine (2-CdA). AB - A technique of protein-DNA flow cytometry was applied to characterize cell cycling, and to assess the cytotoxicity of cladribine (2-chloro-2'deoxyadenosine) toward seven dissociated cultures of human primary brain tumors (anaplastic astrocytoma and glioblastoma multiforme) grown in vitro from surgical biopsies. Control cytograms were suggestive of that a clonogeneic fraction of the cell population consists mainly of cells with low protein content, which do not require increase in protein content before entering the S phase of the cell cycle. Following 24 or 48 hours exposure to cladribine, 1 nM approximately 1 microM, no cytotoxic effect was evident in 4 cultures, whereas in two cases dose dependent progressive block of the phase of the cell cycle was noted. In one case a massive cytotoxic effect resulted in disintegration of culture exposed to 100 nM of the drug. However, the treatment with cladribine was ineffective in a patient bearing the tumor which was the source for the last culture, suggesting that cytotoxicity in vitro may not be predictive of clinical response. PMID- 8705283 TI - Concanavalin A binding by the hippocampal neurons following brief cerebral ischemia in gerbils. AB - Twenty two Mongolian gerbils after 5 min bilateral carotid artery occlusion and 6, 12, 24, 48, 72, 94 hours and 5 days survival were investigated for the neuronal changes in dorsal hippocampus. Paraffin sections were stained with cresyl-violet and marked by their binding of Concanavalin A (Con A) labelled with peroxidase. The degeneration and neuronal loss was observed only in CA1 sectors in almost all experimental groups, whereas the decreased binding of Con A by the neurons of CA1 sector corresponded to the intensity of histologic changes but appeared also only in this sector even without any histological changes. These observation can point at the subthreshold damage of CA1 neurons as result of either diminished supply or increased metabolism of d-glucose, or diminished number of Con A receptors or changes of their specificity after ischemic period. PMID- 8705284 TI - The activity of lysosomal enzymes in visual cortex of rabbits during experimental diabetes. AB - The activity of all principal groups of lysosomal enzymes (acid phosphatase, lipase, beta-galactosidase, sulphatase and cathepsin B) was measured in the visual cortex of rabbits with experimental diabetes. In the first stage of diabetes (21 days), it was observed that enzyme activities in the free fraction and in the membrane-bound fraction are decreased as compared to the initial values determined in healthy animals. In the later stages of diabetes (90-180 days), all lysosomal enzyme activities increased except for sulphatase. This indicated a superiority of catabolic processes in visual cortex cells in the course of experimental diabetes. PMID- 8705285 TI - Stimulation of glucose utilization in the rat claustrum by pain. AB - The involvement of the claustrum in processing of somatosensory information was investigated using the formalin test as an established pain test. The metabolic activity of the claustrum and other brain structures belonging to the somatosensory system was assessed by autoradiographic method of the uptake of (3H)-labelled 2-deoxyglucose (2-DG). Glucose utilization in the claustrum was stimulated by subcutaneous injection of formalin into the front paw. The increase of radioactivity was observed not only in the claustrum, but also in the anterior pretectum, the ventral thalamic nucleus, the somatosensory, insular and piriform cortices on the contralateral side to formalin stimulation. After stereotactical injection of morphine into the contralateral pretectum to stimulation side, uptake of 2-DG in the claustrum was abolished. These results indicate that the claustrum is an important relay nucleus in the somatosensory system, which may convey somatosensory information from the pretectum to the cerebral cortex, thus by-passing the thalamus, throughout the pretecto-claustro-cortical pathways. PMID- 8705286 TI - Antioxidant enzyme activities in different brain areas of the neurological mutant -pt rabbit. AB - Cu, Zn-superoxide dismutase (Cu,Zn-SOD), Mn-superoxide dismutase (Mn-SOD), catalase (CAT), peroxidase (POX), glutathione peroxidase(GP) and glutathione reductase (GR) activities were assayed in the brains of genetically selected neurological mutant rabbits pt and their controls. Paralytic tremor (pt) is a spontaneous mutation in rabbit that affects irregular and defective myelination of CNS. Antioxidant enzyme levels were different in three brain regions: brain hemispheres, cerebellum, and brain stem. In brain hemisphere and cerebellum of pt rabbits Mn-SOD and Cu, Zn-SOD activities were elevated. Catalase activity in brain hemispheres and peroxidase activity in the brain stem of pt rabbits were reduced. It was also noticed, that in the pt rabbit the ratio CAT/Cu, Zn-SOD was lower by 20% in the brain hemispheres and by 13% in the cerebellum and the ratio POX/Cu, Zn-SOD was lower by 31.8% in the brain stem. These findings indicated that pt mutations are associated with changes in the antioxidant defense system in the rabbit brain. PMID- 8705287 TI - Occurrence of lamellar bodies in the rat brain organotypic cultures in normal and pathological conditions. AB - Lamellar bodies (LB) represent an intracytoplasmic organelles of the cells of different origin and various organs. Their exact function is not known. It has been recently suggested that these organelles may play a role in the protein synthesis and regulation of gene expression. Increased number of LB has been demonstrated in differentiating and growing cells. It suggests their functional relation to enhanced metabolism of the cells. Such conditions for cells establish a tissue culture. We have examined organotypic rat brain cultures maintained under standard conditions, and subsequently exposed to anoxia and neurotoxins: kainic and quinolinic acid. In normal conditions LB were occasionally found in neurons. Their number significantly increased in damaged neurons in the experimental conditions. They have also appeared in some damaged astrocytes. We postulate that LB in the brain are connected with cell degeneration. Their origin from rough endoplasmic reticulum is demonstrated. PMID- 8705288 TI - Evaluation of human telencephalon microangioarchitecture in fetal period. AB - The aim of the work was to examine internal vascularization of telencephalon. The brains of 200 fetuses aged 3-8 months of fetal life were examined. Various techniques were used: injection, infrared, Pickworht's method, image computer transformation. Linear and non-linear transformations (mathematical morphology) were also used. The five vascular layers of telencephalon were distinguished: cortical, of white matter, striatal, germinal and of choroid plexus. Their morphology and changes occurring in fetal period were estimated. It was observed that subcortical and paraventricular regions belong to hypovascular areas. White matter vessels were seen as rectilinear without any ramifications. After 6 months of fetal life the most intensive development of inner cerebral vessels occurs. Two age groups were distinguished. The first one comprises the period of 4th to 6th month and is characterized by the existence of long medullary and abundant germinal matrix vessels. The second group (over 7th months) may be described as intensive development of cortical vessels. PMID- 8705289 TI - Electrochemical monitoring of individual exocytotic events from the varicosities of differentiated PC12 cells. AB - Rat pheochromocytoma (PC12) cells have been used as a model of developing neurons to study exocytosis during differentiation. Upon treatment with nerve growth factor, PC12 cells become more neuronal-like. Using amperometric detection at carbon fiber microelectrodes, time-resolved exocytosis of electroactive catecholamines can be observed. The site of exocytosis has been compared for differentiated and undifferentiated cells. Upon differentiation, cells release catecholamines primarily from varicosities along their neurites with no release from the cell body. In addition, the mean vesicular content is not significantly altered upon differentiation although it appears that the distribution of vesicle content becomes more narrow. The number of release events observed also decreases as the cells become more neuronal in character. It is possible that the smaller range of vesicle dopamine content and the decreased number of release events observed after differentiation are a result of the relocation of the site of exocytosis. PMID- 8705290 TI - Acute acidosis elevates malonaldehyde in rat brain in vivo. AB - Oxidative stress in brain tissue was measured experimentally in situ using microdialysis to sample the extracellular environment for a lipid peroxidation breakdown product and antioxidants. The extracellular concentrations of the lipid peroxidation product malonaldehyde (MDA) and the antioxidants ascorbic acid (AA) and uric acid (UA) were measured in rat cortex and striatum in vivo using microdialysis coupled to HPLC with UV detection. Tissue acidosis following ischaemia and epileptic seizures may contribute to neuronal damage, which may be mediated by reactive oxygen species. Perfusion of microdialysis probes with acidic artificial cerebrospinal fluid (pH 6) led to a significant increase in the sampled concentration of MDA and the antioxidant ascorbic acid. Simultaneous perfusion of ascorbate (5 mM) with acidic ACSF (pH 6) completely attenuated the rise in lipid peroxidation. This study provides in vivo evidence for acidosis induced oxidative stress in brain tissue and an antioxidant action of ascorbate. The methodology described here can provide direct in vivo information in respect of oxidative stress in experimental situations. The method could equally be applied to the assessment of oxidative stress in a number of pathological models not necessarily confined to the CNS. PMID- 8705291 TI - Localization of choline acetyltransferase and NADPH diaphorase activities in the submucous ganglia of the guinea-pig colon. AB - A combined immunohistochemical and histochemical demonstration of choline acetyltransferase (ChAT) and nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) was carried out, respectively, to determine the localization of the neurotransmitters, acetylcholine and nitric oxide (NO) in the submucous neurons of guinea-pig colon. Almost half of the submucous neurons in the guinea pig colon exhibited ChAT-immunoreactivity. Some of the ChAT-immunoreactive neurons were also stained for NADPH-d, although most of them showed only weak to moderate diaphorase activity. Many of the submucous neurons displayed exclusively either ChAT or NADPH-d activity. A close spatial relationship was observed between the cholinergic and nitrergic submucous neurons. Thus, in light microscopy, some ChAT-immunoreactive fibres were closely associated with the NADPH-d-positive nerve cell bodies. Ultrastructural study extended the fact that many of the ChAT-immunoreactive terminals made synaptic contacts with the soma of the NADPH-d-positive submucous neurons. A remarkable feature was the demonstration of ChAT and NADPH-d in some of the neurons and their presynaptic axon terminals, suggesting the co-localization of acetylcholine and NO as neurotransmitters in the submucous neurons and their presynaptic axon terminals. It is suggested that the submucous neurons with their specific neurochemical codings would subserve different functions. PMID- 8705292 TI - Neuropeptide Y-immunoreactive primary afferents in the dental pulp and periodontal ligament following nerve injury to the inferior alveolar nerve in the rat. AB - The distribution of neuropeptide Y (NPY)-immunoreactive (IR) primary afferents in the dental pulp and periodontal ligament of the rat mandible were examined following combined chronic constriction injury (CCI) of the inferior alveolar nerve (IAN) and sympathectomy of the superior cervical ganglion (SCG). NPY-IR nerve fibers were observed around the blood vessels in the trigeminal ganglion, dental pulp and periodontal ligament in normal animals. Following combined CCI of the IAN and sympathectomy of SCG (SCGx), perivascular NPY-IR nerve fibers originating from SCG disappeared completely, but many NPY-IR nerve fibers coming from the trigeminal ganglion appeared in the dental pulp and periodontal ligament. In the molar dental pulp, thick NPY-IR nerve fibers were observed within the nerve bundle, and some thin NPY-IR nerve fibers ran towards the odontoblast layer; very few NPY-IR nerve fibers were observed in the incisor pulp. In the periodontal ligament of molar, thick NPY-IR nerve fibers appeared at the alveolar part following combined CCI of IAN and SCGx. In the lingual portion of the periodontal ligament of the incisor, many thick NPY-IR nerve fibers were observed. These occasionally showed a tree-like appearance, resembling immature Ruffini endings; slowly adapting mechanoreceptors. The present results indicate that periodontal mechanoreceptors are among the main targets of injury-evoked NPY following IAN injury. PMID- 8705293 TI - Coincident expression and distribution of melanotransferrin and transferrin receptor in human brain capillary endothelium. AB - One method of iron transport across the blood brain barrier (BBB) involves the transferrin receptor (TR), which is localized to the specialized brain capillary endothelium. The melanotransferrin (MTf) molecule, also called p97, has been widely described as a melanoma specific molecule, however, its expression in brain tissues has not been addressed. MTf has a high level of sequence homology to transferrin (Tf) and lactoferrin, but is unusual because it predominantly occurs as a membrane bound, glycosylphosphatidylinositol (GPI) anchored molecule, but can also occur as a soluble form. We have recently demonstrated that GPI anchored MTf provides a novel route for cellular iron uptake which is independent of Tf and its receptor. Here we consider whether MTf may have a role in the transport of iron across the BBB. The distributions of MTf, Tf and the TR were studied immunohistochemically in human brain tissues. The distributions of MTf and TR were remarkably similar, and quite different from that of Tf. In all brain tissues examined, MTf and the TR were highly localized to capillary endothelium, while Tf itself was mainly localized to glial cells. These data suggest that MTf may play a role in iron transport within the human brain. PMID- 8705294 TI - Reactive microglia specifically associated with amyloid plaques in Alzheimer's disease brain tissue express melanotransferrin. AB - Several investigations have implicated the involvement of metals in neuropathologies. In particular, the disruption of iron metabolism and iron transport molecules have been demonstrated in Alzheimer's disease (AD). We have identified a novel pathway of iron uptake into mammalian cells involving melanotransferrin, or p97, which is independent of the transferrin receptor. Here we investigated whether there is a possible link between this molecule and the pathology of AD. The distributions of melanotransferrin, transferrin and the transferrin receptor were studied immunohistochemically in brain tissues from AD cases. In brain tissues from AD, melanotransferrin and the transferrin receptor were highly localized to capillary endothelium, while transferrin itself was mainly localized to glial cells. In brain tissue derived from AD patients, melanotransferrin was additionally detected in a subset of reactive microglia associated with senile plaques. Our demonstration that melanotransferrin mediates iron uptake through a pathway independent of the transferrin receptor indicates that this mechanism may have a role in AD. PMID- 8705296 TI - The microglial reaction in spinal cords of jimpy mice is related to apoptotic oligodendrocytes. AB - Jimpy is a shortened life-span murine mutant whose genetic disorder results in a severe hypomyelination in the central neruons system associated with a variety of glial abnormalities, including oligodendrocyte death. In this study, we report that oligodendrocyte death in jimpy occurs through an apoptotic mechanism, as demonstrated by in situ labeling of nuclear DNA fragmentation. Compared to those of normal littermates, the spinal cords of jimpy mice showed a significantly higher number of apoptotic cells. Our observations also corroborate that specific glial cell death in jimpy is restricted to oligodendrocytes, as evidenced by double labeling for DNA fragmentation and MBP immunocytochemistry. Cells labeled for DNA fragmentation were always negative for astroglial or microglial markers. Apoptotic oligodendrocytes were not aggregated into clusters and were ubiquitously distributed throughout the jimpy spinal cord, although were more numerous in white matter than in gray matter. We found no physical association between astrocytes and dying cells in jimpy. Microglial cells, however, were found closely attached to and even surrounding apoptotic cells. The possible role of microglial cells in relation to apoptotsis is discussed. PMID- 8705295 TI - The area postrema is involved in paraquat-induced conditioned aversion behavior and neuroendocrine activation of the hypothalamic-pituitary-adrenal axis. AB - Paraquat is a herbicide capable of eliciting conditioned taste aversion (CTA), a behavioral response characteristic of toxicosis. The area postrema (AP) is a hindbrain circumventricular organ previously shown to be important in mediating signs of paraquat-induced toxicity, namely CTA and weight loss. The relationship between neural substrates for paraquat-induced CTA and activation of the hypothalamic-pituitary-adrenal (HPA) axis was investigated in Sprague-Dawley rats with lesions centered on the AP (APX) and sham-operated (SHM) rats administered paraquat (25 mumol/kg) or saline (1 ml/kg). Injection of paraquat at a dose sufficient to condition taste aversion, but produce no other signs of overt toxicity, significantly increased plasma corticosterone concentrations in SHM rats up to 4 h after administration. Paraquat-induced activation of the HPA axis was significantly attenuated in AP-lesioned rats as compared to sham-operated controls. These findings suggest the area postrema is a common neural substrate for the behavioral and neuroendocrine responses to paraquat. PMID- 8705297 TI - Adrenalectomy increases reduced nicotinamide adenine dinucleotide phosphate diaphorase activity in the rat spinal trigeminal subnucleus caudalis. AB - Neurons exhibiting reduced nicotinamide adenine dinucleotide phosphate-diaphorase activity (NADPHd) were quantified at 500 microns rostrocaudal intervals in spinal trigeminal nucleus (Vsp) of adenalectomized (ADX), ADX + corticosterone, and sham ADX rats 6-12 days after surgery. NADPHd neurons were found predominantly in Vsp subnucleus caudalis (Vc) and in dorsomedial subnucleus oralis. ADX significantly increased the number of NADPHd neurons in superficial laminae of Vc, an effect reversed by chronic corticosterone replacement. ADX effects on NADPHd in superficial laminae of Vc but not in deep laminae of Vc or in the periobex region of Vsp paralleled previously observed sites of ADX enhancement of noxious stimulus-induced Fos-like immunoreactivity. The results indicate that chronic changes in adrenal steroid status regulate NADPHd, a mechanism that may both derive from changes in nitric oxide synthase expression and influence the processing of nociceptive information by central trigeminal neurons. PMID- 8705298 TI - Triiodothyronine attenuates estradiol-induced increases in dopamine D-2 receptor number in rat anterior pituitary. AB - Estrogens promote adenohypophyseal enlargement and tumor transformation, and thyroid hormones antagonize these effects. Hormone-induced pituitary enlargement may be mediated by alterations in pituitary dopaminergic function. The present study examined the effects of chronic (20 days) administration of estradiol benzoate (EB), triiodothyronine (T3), or EB and T3 (T3 + EB) on dopamine (D-2) receptors in rat anterior pituitary. D-2 receptor number increased after EB without altered receptor affinity. T3 alone did not affect D-2 receptor number in the anterior pituitary but significantly attenuated the effect of EB. T3 administration also inhibited EB-induced anterior pituitary hyperplasia. D-2 receptor upregulation by EB more likely could reflect a compensatory response to decreased receptor occupation. The present results suggest that D-2 receptors could play an important role in estrogen-induced adenohypophyseal tumor formation and hyperprolactinemia and that thyroid hormones may inhibit estrogen-induced pituitary tumor development via adenohypophyseal D-2 receptors. PMID- 8705299 TI - Regionally specific induction of ICE mRNA and enzyme activity in the rat brain and adrenal gland by LPS. AB - Pro interleukin-1 beta converting enzyme (ICE) activity in the pituitary was found to be significantly increased 4 h after intraperitoneal injection of E. coli lipopolysaccharides, when distribution and inducibility of the enzyme was studied in the adult rat brain and the adrenal gland, using an artificial fluorescence peptide substrate. The same lipopolysaccharide treatment induced ICE mRNA levels in the pituitary, adrenal gland and hypothalamus as studied by reverse transcript-polymerase chain reaction. PMID- 8705300 TI - CREB-1 and CREB-2 immunoreactivity in the rat brain. AB - This study is focused to learn about the cellular localization of transcription factors binding to the cAMP response element-CREBs-in the brain of normal rats and in animals subjected to excitotoxic cell damage. For this purpose, CREB-1 and CREB-2 immunoreactivity is examined in the developing and adult rat brain under physiological conditions, and following systemic kainic acid (KA) injection at convulsant doses in the adult, as a validated experimental model of excitotoxic injury. CREB-1 immunoreactivity is constitutively expressed in periventricular glia and Bergmann glia, and appears in reactive astrocytes following KA-induced excitotoxic cell damage. In contrast, CREB-2 is constitutively expressed in all neurons of the cerebrum, cerebellum and brain stem in the developing and adult brain. CREB-2 immunoreactivity is not increased following KA excitotoxic cell damage. These results demonstrate that CREB-1 and CREB-2 in the brain of the rat are localized in separate cellular compartments and that their expression is differentially regulated in pathologic states. PMID- 8705301 TI - Brief, prolonged and repeated stimuli applied to hyperalgesic skin areas: a psychophysical study. AB - In this experimental study brief/prolonged and single/repeated, nociceptive stimuli (laser, thermode and electrical) were used to investigate sensory changes in capsaicin-induced primary and secondary hyperalgesia. The pain threshold to prolonged thermode stimulation was reduced in the primary area and remained constant in the secondary area. The pain thresholds to brief laser and electrical stimuli remained constant in the primary but reduced in the secondary area. The summation pain threshold to repeated (five stimuli delivered at 0.5, 1, 2 and 3 Hz) laser and electrical stimuli was reduced in the secondary area. The stimulus response functions to single laser and electrical stimuli were increased in the secondary area. PMID- 8705302 TI - Nitric oxide synthase-containing neurons in the hippocampus are preserved in trimethyltin intoxication. AB - We studied the effects of trimethyltin (TMT) (9 mg/kg, p.o.) on the nitric oxide synthase (NOS)-containing neurons in the rat hippocampus by NADPH-diaphorase histochemistry and a biochemical assay of NOS activity. TMT exposure caused the typical behavioral changes and a loss of the CA3/4 pyramidal cells, which were NADPH diaphorase-negative. The scattered interneurons and the CA1 pyramidal cells, which were NADPH diaphorase-positive, were spared. Hippocampal NOS activity showed no reduction in the TMT-treated rats compared with the controls. These results provide evidence of the preservation of the NOS-containing neurons in TMT intoxication. PMID- 8705303 TI - Delayed kindling development after rapidly recurring seizures: relation to mossy fiber sprouting and neurotrophin, GAP-43 and dynorphin gene expression. AB - Development of kindling and mossy fiber sprouting, and changes of gene expression were studied after 40 seizures produced during about 3 h by electrical stimulation every 5 min in the ventral hippocampus. As assessed by 5 test stimulations, enhanced responsiveness was present already after 6-24 h but from 1 week post-seizure increased gradually up to 4 weeks without additional stimuli. Sprouting of mossy fibers in the dentate gyrus was demonstrated only at 4 weeks with Timm's staining. In situ hybridization showed a transient increase (maximum at 2 h) of brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF), TrkB and TrkC mRNA levels and reduction (maximum at 12-24 h) of neurotrophin-3 (NT-3) mRNA expression in dentate granule cells after the seizures. In addition, BDNF mRNA levels were elevated in CA1 and CA3 regions, amygdala and piriform cortex. Marked increases of mRNA for growth-associated protein (GAP-43), with maximum expression at 12-24 h, were observed in dentate granule cells and in amygdala-piriform cortex. Dynorphin mRNA levels showed biphasic changes in dentate granule cells with an increase at 2 h followed by a decrease at 24 h. No long-term alterations of gene expression were observed. These findings indicate that increased responsiveness develops rapidly after recurring seizures but that the kindled state is reached gradually in about 4 weeks. Mossy fiber sprouting occurs in parallel to epileptogenesis and may play a causative role. Short-term changes of neurotrophin and Trk, GAP-43 and dynorphin mRNA levels and the assumed alterations of the corresponding proteins could trigger structural rearrangements underlying kindling but might also contribute to the initial increase of seizure susceptibility. PMID- 8705304 TI - Coexistence of nitric oxide synthase and neuropeptides in the mouse vomeronasal organ demonstrated by a combination of double immunofluorescence labeling and a multiple dye filter. AB - Nitric oxide synthase (NOS)-immunofluorescence techniques were applied to the mouse vomeronasal organ. Immunoreactivity for NOS was found in the nerve fibers distributed in the receptor-free epithelium, and around the blood vessels and glands in the cavernous tissue. No NOS fibers were seen in the receptor area. A combination of double immunofluorescence labeling and multiple dye filter revealed that a part of the substance P (SP)-immunoreactive nerve fibers in the cavernous tissue contained NOS and that all the vasoactive intestinal polypeptide (VIP)-immunoreactive nerve fibers around the blood vessels and glands in the cavernous tissue contained NOS. A few SP-immunoreactive cell bodies in the trigeminal ganglion showed coexistence with NOS, and almost all VIP immunoreactive cell bodies in the sphenopalatine ganglion showed coexistence with NOS. Immunoreactivity for NOS without VIP in the cell bodies in the sphenopalatine ganglion was also found. These results suggest that NOS immunoreactive nerve fibers in the mouse vomeronasal organ originate from the trigeminal and the sphenopalatine ganglia, and may modulate the vascular tone and the glandular secretion. In addition, these functions may be controlled in part by the interaction of nitric oxide and neuropeptides. PMID- 8705305 TI - Nonuniform distribution of cell proliferation in the adult teleost retina. AB - Teleost fish continue to grow throughout life, and their eyes enlarge correspondingly. Within the eye, the retina grows by stretching existing tissue and adding new cells. Cell addition occurs in two ways: First, all cell types except rod photoreceptors are added circumferentially at the edge of the eye where the retina meets the iris; second, rod photoreceptors are generated from a population of rod progenitor cells which divide throughout the outer nuclear layer (ONL). To determine the spatial distribution of rod progenitor cells across the teleost retina, we labeled dividing cells with an antibody to proliferating cell nuclear antigen (PCNA) throughout a 24 h period. We found a significantly higher density of dividing rod precursor cells at the nasal and temporal margins than in the central retina throughout the 24 h cycle. At night, the density of dividing cells is significantly greater at the nasal pole of the eye. The difference between cell division at the center and the margin was reduced at night when the density of cell division in the central retina increased significantly. Taken together, these data suggest that the eye grows asymmetrically, with more cells added at the nasal pole. Possible developmental causes and functional consequences of the reported distribution of cell divisions in time and location are presented. PMID- 8705306 TI - The neurotrophin NT4/5, but not NT3, enhances the efficacy of nigral grafts in a rat model of Parkinson's disease. AB - The neurotrophins NT4/5 and NT3 have previously been shown to improve the survival and fibre outgrowth of embryonic dopaminergic neurons in vitro. In the present study we attempted to augment the efficacy of embryonic nigral grafts in vivo. This was done by directly infusing the neurotrophins intraparenchymally in close proximity to transplanted nigral tissue placed in the dopamine depleted striatum of 6-hydroxydopamine lesioned rats. Our results indicated that NT4/5, but not NT3, stimulated fibre growth from embryonic nigral cells and enhanced functional efficacy of the grafts as assessed by metamphetamine-induced rotation. PMID- 8705307 TI - The effects of immunolesions of nerve growth factor-receptive neurons by 192 IgG saporin on sleep. AB - Low-affinity nerve growth factor (NGF) receptors are present on the cholinergic neurons of the basal forebrain. We studied the effects of 192 IgG-saporin, a specific immunotoxin for the NGF receptor-positive, cholinergic basal forebrain neurons, on sleep, the power spectrum of the electroencephalogram (EEG), and body temperature. After 3 d baseline recordings, 12 male rats were injected intracerebroventricularly with 4 micrograms 192 IgG-saporin. EEG, motor activity, and brain temperature were recorded for 23 h on the first, third, fifth, and seventh day after the treatment. 192 IgG-saporin did not affect the total daily amounts but altered the circadian distribution of sleep. On days 1 and 3 after the injection of the immunotoxin, the amount of non-rapid-eye-movement sleep (NREMS) and rapid-eye-movement sleep (REMS) increased during the dark period, whereas during the light both NREMS and REMS decreased. On day 5, these changes were less pronounced and sleep completely returned to the baseline by day 7. The EEG was suppressed in each frequency band and each vigilance state, and, in contrast to sleep, these changes in EEG persisted for 7 days. Brain temperature was decreased from day 3. These results suggest that NGF receptor-positive, cholinergic basal forebrain neurons are not necessary for the maintenance of total sleep time but contribute to the generation of normal EEG and the maintenance of brain temperature. PMID- 8705308 TI - Long-term time course of regional changes in cholinergic indices following transient ischemia in the spontaneously hypertensive rat brain. AB - Using an animal model of forebrain ischemia in spontaneously hypertensive rats (SHR) by 3-h bilateral carotid occlusion, and various indices of the cerebral cholinergic system were assessed for periods up to 24 weeks. The lesions observed histologically in the hippocampus of SHR 2 weeks after ischemia were less severe than those in the frontal cortex. Marked elevation of acetylcholine concentration was transiently observed in the frontal cortex, hippocampus and thalamus + midbrain at 2 weeks, and in the striatum at 1-4 weeks after ischemia. Choline acetyltransferase activity remained unchanged in all regions throughout the experimental period except for a minimal decrease in the frontal cortex at 4 weeks. Choline esterase (ChE) activity was slightly decreased in the frontal cortex at 2-4 weeks after ischemia but recovered by 8 weeks. A decrease in the hippocampus was seen at 8 weeks. The B(max) for the M1-receptor was significantly reduced by 2 weeks in the frontal cortex and by 4 weeks in the hippocampus. Low B(max) values in both regions persisted through week 24. These delayed hippocampal changes in the ChE activity and M1-receptor in SHR were similar to those of the very much delayed changes in M1-receptor previously reported in the gerbil model for transient ischemia. In contrast, Wistar-Kyoto rats (WKY), used as normotensive controls, exhibited no histological or biochemical changes for up to 24 weeks. The difference between SHR and WKY may depend on the more severe cerebral blood flow depletion during carotid ligation in the former. The chronic state of SHR after the transient ischemia may be a useful pathophysiological model for human cerebral infarctions with hypertension. PMID- 8705309 TI - Testosterone-dependent increase of gap-junctions in HVC neurons of adult female canaries. AB - Singing of canaries is controlled by a chain of interconnected brain areas. One of these areas, the caudal nucleus of the ventral neostriatum (HVC), is sensitive to androgens and estrogens. In adult canaries, both male and female sing. Female song is structured differently from the male singing and characterized by a higher variability. Singing females were implanted with testosterone propionate (n = 5) or with empty silastic tubes (n = 5). Testosterone treated females developed a male-like song and had an increased number of neuronal soma-somatic gap junctions in the HVC compared with the untreated singing females. Electric coupling of HVC neurons could be important for the testosterone-dependent changes of the song pattern of canaries. PMID- 8705311 TI - GABAergic inhibitory response of locus coeruleus neurons to caloric vestibular stimulation in rats. AB - We examined the effects of caloric vestibular stimulation on the neuronal activity of the locus coeruleus (LC) in urethane-anesthetized rats. The middle ear cavity was irrigated with hot (44 degrees C) or cold (30 degrees C) water through a polyethylene tube. Most neurons (hot water: 76%, 55/72; cold water: 90%, 19/21) exhibited suppression of neuronal discharge in response to caloric stimulation. The suppression of LC neuronal discharge following caloric stimulation occurred with a long latency (approximately 80 s), and lasted a long period of time (approximately 3 min). Neither caloric stimulation of the auricle, nor irrigation of the middle ear with water at 37 degrees C, nor caloric stimulation of the middle ear after labyrinthectomy inhibited LC neuronal discharge. The caloric stimulation-induced LC neuronal inhibition was significantly attenuated by the intravenous injection of picrotoxin and by the iontophoretic application of bicuculline methiodide. These findings indicate that the predominant effect of caloric vestibular stimulation on LC neuronal discharge is inhibitory, and that the caloric stimulation-induced LC neuronal inhibition is mediated by GABAA receptors located on the membrane of LC neurons. It is suggested that the suppressed activity of noradrenergic LC neurons is involved in the vestibulo-autonomic reflex. PMID- 8705310 TI - Glycine and GABAA receptor-mediated chloride fluxes in synaptoneurosomes from different parts of the rat brain. AB - Strychnine-sensitive, inhibitory glycine receptors have not until lately been considered to play a significant role in neurotransmission in mammalian forebrain regions. In order to investigate the role of glycine as a neurotransmitter in brain we have measured glycine induced chloride fluxes in different adult rat forebrain areas using synaptoneurosomes and a chloride-sensitive fluorescent indicator. The results have been compared to those obtained with GABA. The synaptoneurosomes from every brain area investigated responded to both glycine and GABA with chloride fluxes in a picrotoxin sensitive manner. The effect of glycine was inhibited by strychnine, which had no effect on the GABA-induced Cl flux. Bicuculline inhibited the effect of GABA, but had no effect on the glycine induced Cl-flux. Addition of GABA did not affect the response to glycine and vice versa. The endogenous content of glycine and GABA in the synaptoneurosome preparations was about the same and synaptoneurosomes from every brain area investigated released both glycine and GABA upon depolarisation with KCl. The depolarisation induced release of both GABA and glycine was partly Ca(2+) dependent and partly Ca(2+)-independent. These results indicate that glycine can induce inhibitory Cl- fluxes distinct from GABA induced fluxes in every investigated brain area and that glycine can be released upon depolarisation. PMID- 8705312 TI - Short-photoperiod exposure reduces L-aromatic-amino-acid decarboxylase immunostaining in the arcuate nucleus and median eminence of male Syrian hamsters. AB - In male hamsters, exposure to short photoperiod (SD) results in a decrease in median eminence and posterior pituitary dopamine (DA) concentrations. To determine if the SD-induced decline in DA is due to a decrease in the number of neurons synthesizing DA, immunocytochemistry was used to identify cells containing tyrosine hydroxylase (TH) and L-aromatic amino acid decarboxylase (AADC) in animals housed under long photoperiod (LD) or SD. Immunopositive cells were counted in the arcuate and caudal periventricular nuclei, regions where the DA neurons that project to the median eminence and posterior pituitary are located. AADC immunopositive (AADC +) cells were also counted in the median eminence. Photoperiod did not affect the number of TH immunopositive (TH+) or AADC + cells in the caudal periventricular nuclei. In the arcuate nuclei, SD exposure did not affect the number of TH+ neurons, but produced a significant decline in the number of AADC + cells. The number of AADC + cells was also reduced in the median eminence of SD-exposed animals. This decline in AADC+ cells may play a role in SD-induced changes in hypothalamic regulation of anterior pituitary hormone release, including the decline in median eminence DA concentrations. PMID- 8705313 TI - Effects of central chemical drive on poststimulatory respiratory depression of laryngeal origin in the adult cat. AB - We investigated the influences of central CO2-related chemosensory drive on poststimulatory respiratory phenomena induced by superior laryngeal nerve (SLN) stimulation in pentobarbitone-anesthetized, vagotomized, carotid sinus denervated, paralyzed, and artificially ventilated adult cats. Respiratory output was monitored as integrated phrenic nerve activity. Under eucapnic conditions, apnea-producing SLN stimulations of both short (10 s) and long (30 s) duration were followed by persistent apnea and depression in phrenic motor output; the latter showed a gradual recovery that followed an exponential time course. Hypocapnia increased the duration of poststimulatory apnea and the intensity of poststimulatory depression in phrenic minute output owing to changes in peak phrenic activity. Hypercapnia did not affect the duration of poststimulatory apnea, but markedly attenuated poststimulatory depression in respiratory activity, mainly due to changes in respiratory frequency. The rate of respiratory recovery was similar under eucapnic and hypocapnic conditions, but it was slower during hypercapnia. The results provide evidence that central chemosensitivity plays a prominent role in counteracting poststimulatory depressant effects on respiration induced by SLN stimulation. PMID- 8705314 TI - Low doses of neurotensin in the preoptic area produce hyperthermia. Comparison with other brain sites and with neurotensin-induced analgesia. AB - High amounts of neurotensin (NT) are found in the preoptic area of the hypothalamus, an area known to be involved in the regulation of body temperature. It is generally believed that NT is a peptide that produces hypothermia, and several sites in the brain have been proposed to mediate NT-induced hypothermia, including the preoptic area. However, the doses of NT used in these experiments were always very high (microgram order) whereas, according to Goedert, the total brain content of NT in the rat does not exceed 10 ng. We therefore reinvestigated the effects of microinjections of NT in the brain, using high (5 micrograms) and low (50 and 5 ng) doses, into the preoptic area and other brain sites (cerebral ventricles, posterior hypothalamus, and nucleus accumbens), and we also studied, as a comparison, the effects of high and low doses of NT on pain sensitivity in the same sites. The results show that the preoptic area has unique properties in the regulation of body temperature: low doses of NT in the preoptic area produce a hyperthermic response, whereas high doses produce hypothermia. In comparison, NT produces hypothermia in the posterior hypothalamus whatever the dose, and NT has analgesic effects in the preoptic area only at high doses. Besides, NT has no thermic effect, but does have an analgesic effect, in the nucleus accumbens. The selectivity of the actions of high doses of NT, as well as the mechanism of action of NT (possibly an endogenous neuroleptic), are discussed. PMID- 8705315 TI - Testosterone effects on the neuronal ultrastructure in the medial preoptic nucleus of male Japanese quail. AB - Dorsolateral neurons of the medial preoptic nucleus (POM) of male Japanese quail are sensitive to the plasma levels of testosterone: their volume and optical density in Nissl-stained sections increase in castrated birds treated with testosterone. The present study was performed on castrated male quail treated or not with Silastic implants filled with testosterone to describe the ultrastructural variations induced by testosterone in these neurons. Gonadally intact male birds were included as controls. The ultrastructure of neurons, taken from the dorsolateral portion of the POM, was dramatically affected by the endocrine manipulations. Quantitative evaluations demonstrated a significant decrease in castrated birds of the rough endoplasmic reticulum (RER), of free polyribosomes, of Golgi complexes, and of dense bodies; these changes paralleled the decrease in cell size. The cell size and the percentage of volume occupied by the intracellular organelles in castrated birds treated with testosterone were comparable to values observed in controls. These ultrastructural changes are similar to those observed in neuronal targets for other gonadal hormones, supporting the idea that testosterone stimulates the development of cytoplasmic structures involved in protein synthesis and secretion. In addition, exposure to testosterone affects the synaptic inputs to POM. These ultrastructural changes are presumably related to the physiological effects (e.g., activation of male sexual behavior) exerted by testosterone on this preoptic region. PMID- 8705316 TI - Bilateral lesions of the hypothalamic suprachiasmatic nucleus eliminated sympathetic response to intracranial injection of 2-deoxy-D-glucose and VIP rescued this response. AB - We previously found that bilateral lesions of the suprachiasmatic nucleus abolished hyperglycemic response to intracranial injection of 2-deoxy-D-glucose in rats. Because the hyperglycemia due to 2-deoxy-D-glucose was shown to be dependent on the functions of the adrenal medulla and sympathetic nervous system, the effect of bilateral lesions of the suprachiasmatic nucleus on changes in the nervous activity of sympathetic efferents to the adrenal after intracranial injection of 2-deoxy-D-glucose was examined in rats. It was found that bilateral lesions of the nucleus eliminated the increase in neural activity of the sympathetic efferent that occurred after the injection of 2-deoxy-D-glucose. Because the suprachiasmatic nucleus possesses neurons containing a vasoactive intestinal polypeptide-like substance, the effects of vasoactive intestinal polypeptide and 2-deoxy-D-glucose, administered alone or in combination, on the sympathetic activity were examined in intact control rats and in rats with bilateral lesions of the suprachiasmatic nucleus. It was found that in the normal control rats, vasoactive intestinal polypeptide alone increased the sympathetic activity, whereas it dramatically enhanced the sympathetic response to 2-deoxy-D glucose. However, in rats with bilateral lesions of the suprachiasmatic nucleus, vasoactive intestinal polypeptide alone elicited no increase in the nervous activity of the sympathetic efferents to the adrenal, but combined administration of vasoactive intestinal polypeptide and 2-deoxy-D-glucose caused an increase in the nervous activity of sympathetic efferents to the adrenal. These findings suggest that the suprachiasmatic nucleus is involved in the enhancement of sympathetic activity caused by intracranial injection of 2-deoxy-D-glucose, and that neurons containing a vasoactive intestinal polypeptide-like substance in the suprachiasmatic nucleus play an important role in the sympathetic enhancement that occurs after intracranial injection of 2-deoxy-D-glucose. This role might be a permissive and facilitative one. PMID- 8705317 TI - Beta-amyloid1-40 inhibits electrically stimulated release of [3H]norepinephrine and enhances the internal calcium response to low potassium in rat cortex: prevention with a free radical scavenger. AB - We examined the effects of beta-amyloid1-40 (A beta 1-40) on the electrically stimulated release of [3H]norepinephrine (NE) and the potassium chloride (KCl) evoked increases in intrasynaptosomal calcium ([Ca2+]i) from rat brain cortical slices and synaptosomes, respectively. The results show that 3nM A beta 1-40 decreased electrically stimulated [3H]NE release approximately 50%, and the same concentration of A beta 1-40 increased [Ca2+]i by 78% above control during low level (10 mM) KCl depolarization. Complete prevention of both effects was observed when the free radical scavenger 1mM N-tert-butyl-alpha-phenylnitrone (PBN) was present. Also, when PBN was exposed to the slice first and then followed by A beta 1-40, the inhibition of [3H]NE was blocked. Alone, PBN had no effect in either paradigm. Our experiments show that 3 nM A beta 1-40 allows a PBN-sensitive free radical to deleteriously effect both evoked [3H]NE release and [Ca2-]i regulation in rat cortical slices and synaptosomes. PMID- 8705318 TI - Hypothalamic serotonin release and raised blood pressure after raphe nuclei stimulation in rats. AB - Electrical stimulation of dorsal raphe (DR) nuclei, in addition to enhancing serotonin (5-HT) release in the hypothalamus, elicited proportional hypertension and tachycardia in anesthetized rats. This could be mimicked by microinjection of two excitatory amino acids, kainic acid and glutamate, into the DR of rat brain. Intrahypothalamic administration of DOI (a 5-HT2 agonist), but not 8-OH DPAT (a 5 HT1 agonist) or 2-methyl-serotonin (a 5-HT3 agonist), also produced both hypertension and tachycardia in rats. The DR stimulation-induced hypertension, tachycardia, or increased hypothalamic 5-HT release were attenuated by prior destruction of the ascending serotonergic system produced by ICV injection of 5,7 dihydroxytryptamine and by prior blockade of postsynaptic serotonergic receptors produced by intrahypothalamic injection of 5-HT2 antagonists, cyproheptadine and ketanserin. The DOI-induced hypertension and tachycardia were also reduced by prior blockade of 5-HT2 receptors with cyproheptadine or ketanserin. Thus, it appears that DR stimulation activates the 5-HT release in the hypothalamus, then activates the hypothalamic 5-HT2 receptors and results in both hypertension and tachycardia in rats. PMID- 8705319 TI - Recovery of function after brain damage: differential effects of blocking calcium uptake during the recovery of a learned behavior and the performance of the recovered behavior following neocortical brain injury. AB - The recovery of a learned behavior following brain damage is typically considered to have occurred when the brain-injured individual reattains the performance criterion that defined original preoperative learning. While this is obviously correct from an operational point of view, it does not necessarily mean that the consequences of the brain injury have been reversed, particularly with regard to the sustained performance of the supposedly recovered behavior. The present research attempted a more comprehensive evaluation of the behavioral effects of localized neocortical injury by investigating how a calcium channel blocker would effect (a) the original preoperative acquisition, (b) the initial postoperative recovery, and (c) the subsequent long-term performance of a brightness discrimination learned by rats subjected to injuries of their visual neocortex. The results demonstrated that notwithstanding the brain-injured rat's ability to reattain the performance criterion used to define preoperative learning, its long term performance of this recovered behavior was significantly inferior to that of a normal rat. More importantly, the present data suggest that there are important differences between the initial postoperative recovery of a behavior and its long term performance since the same drug that will facilitate initial recovery has just the opposite effect with respect to the animals long-term postoperative performance of the behavior. PMID- 8705320 TI - Effect of deprivation of serotonin by p-chlorophenylalanine on induction and maintenance of pseudopregnancy in female rats. AB - The effect of serotonin synthesis inhibitor, p-chlorophenylalanine (PCPA), on induction and maintenance of pseudopregnancy as indicated by deciduoma formation was examined in female rats. Animals were injected with 1 mg/kg b.wt. of reserpine on the day of metestrus, and silk thread was passed through and placed in the left uterine horn 3 days after reserpine to induce deciduoma. PCPA (100 mg/kg b.wt.) was injected daily for 4 days before or after reserpine in 15 and 13 rats, respectively. A single injection of PCPA was administered before reserpine in nine females. In another group of rats (N = 16), instead of PCPA, saline was injected four times before reserpine. Nineteen female rats were treated with reserpine only as a control group. Results showed 89% of the control and 81.3% of the saline-treated females had massive deciduoma in traumatized uterine horn. In contrast, only 33.3% or 46.2% females with daily treatments of PCPA for 4 days before or after reserpine showed positive decidual reaction. In addition, 88.9% of females with single injection of PCPA possessed uterine horns with deciduoma. These results suggest that 4 days of treatment with PCPA eliminate induction and/or maintenance of pseudopregnancy. Thus, some levels of serotonin are required to induce and maintain pseudopregnancy. PMID- 8705321 TI - [Functional and morphological changes in the respiratory organs of cats after long-term exposure to pure normobaric oxygen]. AB - BACKGROUND: Pulmonary oxygen toxicity is very well known and proved. The influence of hyperoxia on the respiratory reflexes is not known till now. MAIN PURPOSE: To ascertain if long-lasting breathing of pure normobaric oxygen (PNO) alter respiratory reflexes. METHODS: 34 adult cats of both sexes, weighing 2.5 4.0 kg, were used in two experiments. In the first experiment 16 animals with inserted chronic tracheal cannula (CTC) were used. In the second experiment 18 animals without CTC were employed. Part of animals in both experiments was exposed to PNO (day by day for 2 weeks, 10 h daily), remaining animals were exposed to room air under the same conditions. Side tracheal pressure was recorded in unanesthetized animals of the first experiment. The second experiment was performed in anaesthetised animals (Pentobarbital Spofa, 35 mg/kg, i.p.). Oesophageal pressure and blood pressure in femoral artery were recorded. Cough reflex, sneezing and aspiration reflexes were induced by mechanical stimulation of airway mucosa. Pulmonary chemoreflex was elicited by i.v. administration of 50 micrograms phenyl biguanid. Hering-Breuer inflation reflex was induced by lung inflation with pressure of 1 kPa. Reactivity of tracheal and pulmonary smooth muscle to histamine were measured in vitro. Differences in recorded parameters between animals exposed to PNO, and to room air, were tested by Mann-Whitney Wilcoxon test and by Student's t-test. When p < 0.05, the differences were recognized as significant. RESULTS: Significant decreasing of the expiratory parameters of the cough induced from laryngopharyngeal mucosa, inhibition of sneezing, and inhibition of aspiration reflex, were found in animals exposed to PNO. Relaxing reaction of tracheal smooth muscle of control animals to histamine was reversed to contraction in animals exposed to PNO. Morphological changes of the respiratory tract induced by influence of oxygen were found CONCLUSION: Long lasting breathing of PNO induced changes of respiratory reactions elicited mainly from upper airway.(Fig. 6, Tab. 3, Ref. 22) PMID- 8705322 TI - [Cardiovascular and hemodynamic changes after artificial pulmonary ventilation]. AB - The cardio-respiratory interactions include mechanical, reflex and humoral mechanisms. However, in the organism they mutually overlap, thus mating their separate investigation is problematic. Mechanical effects of conventional artificial ventilation (AV) and high frequency ventilation (HFV) are elicited by increased intrapulmonary pressure during lung inflation, as well as during application of positive end expiratory pressure (PEEP). The increase in intrapulmonary pressure compresses the pulmonary vessels and increases the pulmonary vascular resistance. These changes deteriorate the right ventricular function. Mechanical factors are responsible for the fall of the left ventricular (LV) filling, stroke volume and alteration of the LV preload and afterload. LV filling is decreased during artificial ventilation (AV) owing to the shifting of blood from the central to the peripheral circulation, and to the rise in pulmonary vascular resistance. Application of positive intrathoracic pressure during ventricular ejection phase can diminish LV afterload and increase the cardiac output. This effect is striking in LV failure. The reflex effects of artificial ventilation are due to the activation of baroreflexes and pulmo vagally cardiac and vasoactive reflexes. Activation of these reflexes depends on the level and characteristic of the pressure in the airways, lungs, heart and vessels. Humoral effects of AV on the cardiovascular system and hemodynamics are triggered by lung expansion, circulatory changes, and they result in a release of vasoactive substances from lung parenchyma. (Fig. 5, Ref. 45.) PMID- 8705323 TI - [Selected parameters of lipoprotein metabolism in cerebrovascular diseases]. AB - In series of patients with stroke, selected by random (n = 68), mean age 62.44 +/ 9.12 years (range 39-82 yrs), there were 23 females (33.8%), mean age 65.43 +/- 10.11 yrs and 45 males (66.2%) mean age 60.8 +/- 8.3 yrs. Lp(a) reference values have been obtained from a group of 283 healthy individuals (age ranging from 15 to 65 years). The cholesterol, triacyglycerol, Apo B reference values come from the database of the Department of Clinical Biochemistry. There were 52 hypoxemic stroke patients in the whole observed group. Triacylglycerol serum level TAG < or = 2.89 mmol/l was observed in 47 cases (90.3%), the serum level TAG > 2.89 mmol/l was present in 5 cases (9.7%). The occurrence of TAG normal serum level was significantly more frequent than its pathologic increase (p < 0.001). Apolipoprotein Apo B < or = 1.67 g/l serum level was present in 41 (78.8%) and Apo B > 1.67 g/l in 11 (21.2%) cases (p < 0.001). Apo B < or = 1.67 g/l serum levels in 23 cases (82.1%) and Apo B > 1.67 g/l in 5 cases (18%) were observed among the stroke diabetes mellitus patients (n = 28)--statistic difference in 1/1000 level. In the total hypoxemic stroke group (n = 52), Lp(a) < or = 0.278 g/l was observed in 44 cases (84.6%), Lp(a) > 0.278 g/l serum level was present in 8 cases (15.4%)/ - p < 0.001. According to EASD consensus the serum level of Lp(a) = 0.278 g/l has been considered as "cut-off limit". Similar distribution of Lp(a) serum levels was observed in the diabetes mellitus stroke group (n = 28), the ischemic heart group (n = 54), the group with aortosclerosis (n = 16) and in the group with arterial hypertension (n = 50). Elevated TAG serum levels were not in correlation with the number of sites where atherosclerotic changes were proved by arteriography, ultrasound investigation e.g. in the extracranial brain supplying arteries. Elevated Lp(a) serum levels did not correlate with the stage of ischemic heart disease and they correlated with the stage of functional CNS defect in arterial hypertension and atherosclerosis. Metabolic disorders of lipoprotein and apolipoprotein, namely genomic transcription of lipoprotein seem to be more significant risk stroke factors, but, if they are present, they contribute to the occurrence of arteriosclerosis of some larger arteries. Elevated Lp(a) serum levels did not correlate with the stage of the heart ischemic disease and aortosclerosis, but they correlate with the stage of functional CNS defect due to arteriosclerosis and arterial hypertension, hence the increase in Lp(a) serum level as an indicator of arteriosclerotic evolution of cerebral arteries is significant. Our results, hence, do confirm a common supposition for Lp(a) serum level as an independent arteriosclerotic risk factor of the brain arteries. (Fig. 7, Tab. 1, Ref. 22.) PMID- 8705324 TI - [Can permanent sinus arrhythmia in conscious dogs be suppressed with anesthesia?]. AB - OBJECTIVE: Large prospective studies in dogs with healed myocardial infarction (MI) recently show a positive relation between heart rate variability (HRV) and sudden cardiac death. METHODS: We have done similar experiments in dogs and we studied HRV, ECG and body surface potential maps (BSPM) in 26 normal mongrel dogs (10-15 kg) and 12 dogs with an experimental MI (ligation of LAD). A two-channel 8 hours lasting ECG recording was performed in all conscious dogs. The 2nd recording was done in Penthobarbital anaesthesia (30 mg/kg). RESULTS: We have found sinus arrhythmia (SA) in all 26 conscious dogs. The anaesthesia suppressed the sinus arrhythmia and HRV via compensatory tachycardia and alterations in baroreflexes. The suppression of arrhythmia was also present in dogs with myocardial infarction. CONCLUSIONS: It is suggested, that HRV and SA in dogs depend on conscious state and anaesthesia. On the basis of our results we can anticipate that the most important for HRV is the present status of the sympathetic nerve. We suggest, that our results are an important finding for experimental arrhythmology. (Fig. 4, Ref. 24.) PMID- 8705325 TI - [Extrauterine choriocarcinoma--a rare form of gestational trophoblastic disease]. AB - Choriocarcinoma represents the most serious form of trophoblast gestation disease. In the majority of cases the carcinomatous tissues fill out the uterine cavity, or they grow in a form of nodes deep in the uterine wall. The primary extrauterine localization of this tumour is very rare. The authors describe two cases of choriocarcinomas with tubal or ovarian localization. (Fig. 4, Ref. 19.) PMID- 8705326 TI - [The effect of experimental gastroesophageal reflux on the cough reflex in anesthetized cats]. AB - BACKGROUND: Gastroesophageal reflux (GER) is a common cause of chronic cough. It has been suggested that GER most often causes chronic cough by stimulating the distal oesophagus. Possible mechanisms of this interactive process are: a) an oesophageal-tracheobronchial reflex, b) acid reflux seems to be only a cofactor of cough; it decreases cough threshold. AIM: To evaluate the effects of experimental GER on the cardiorespiratory parameters and on the cough induced by mechanical stimulation of airways mucosa in anaesthetised cats. METHODS: In 10 adults cats of either sex, mean body weight 2.4 +/- 0.2 kg, anaesthetised with pentobarbitone sodium (30 mg/kg b.w., i.p.), oesophageal pressure, blood pressure, and volume tidal were recorded. 2 ml 0.1 N HCl was instilled into the isolated thoracic oesophagus with preserved innervation over a period of 5 minutes. The cough was induced by the insertion of nylon fibre into the airways and its intensity was evaluated from the changes in oesophageal pressure. Cough challenge was also repeated during intraesophageal instillation of 2 ml capsaicin (350 mumol/l). The parameters of cough intensity elicited during experimental GER were compared with the control cough parameters obtained during intraesophageal instillation of 2 ml physiological saline. RESULTS: Intraesophageal instillation of either HCl or capsaicin was accompanied by the contractions of oesophageal wall and by the fluctuations in volume tidal. These changes were more pronounced after capsaicin challenge. There was no induction of either cough or other respiratory reflexes with forced expiration during the experimental GER. In other part of this study there were not obtained any significant differences of the intensity of mechanically-induced cough during intraesophageal instillation of physiological saline, HCl, and capsaicin. CONCLUSION: The oesophageal tracheobronchial reflex mechanism for either induction or modulation of cough is not present in healthy anaesthetised cats. We agree with previous data that GER alone is not trigger agent of cough. (Fig. 5, Ref. 17.) PMID- 8705327 TI - [High-resolution electrocardiography in monitoring myocardial damage after therapy with anthracyclines in children]. AB - BACKGROUND: Anthracycline cytostatics, widely used in oncologic practice, may induce discrete myocardial damage occasionally culminating in life-threatening cardiologic complications. The most serious clinical manifestations of anthracycline cardiotoxicity are dilated cardiomyopathy, heart failure and fatal arrhythmias. OBJECTIVES, STARTING POINT AND MAIN PURPOSE: High-resolution electrocardiography (HRECG) is one of the latest cardiologic methods, which can be promising for early identification of patients at risk of anthracycline cardiotoxicity. The aim of this study is the evaluation of the incidence of HRECG abnormalities in a group of paediatric patients treated with anthracyclines and the usefulness of HRECG for stratification of patients at risk of the clinical cardiotoxicity. PATIENTS AND METHODS: A set of 60 oncologic paediatric patients treated with anthracyclines was divided into two groups. The first group was formed by 15 patients undergoing evaluation during their anthracycline therapy (median after the last administration of antracyclines was 3.2 days). Their average age at the time of examination was 14.7 +/- 4.1 years. The total cumulative dose of antracyclines was 40-300 mg/m2 (median 150 mg/m2). The second group was formed by 45 patients who were evaluated after completing anthracycline therapy. The interval of time from the last administration of antracycline in this subgroup of patients was 3 months-12 years (median 5.5 years). Their average age at the time of HRECG examination was 14 +/- 4.1 years. The total cumulative dose of anthracyclines was 90-440 mg/m2 (median 230 mg/m2). Six patients of this group (13.3%) were treated also with mediastinal radiotherapy (18-40 Gy). 43 patients (95.5%) of second group were in complete remission, two other patients yielded a progression of their malignancy. 10 patients (22%) were examined by HRECG 2-5 times in app. two-month intervals. The control group was formed by 30 randomly selected healthy children and adolescents with normal ECG. Average age was 15.1 +/- 5.8 years. Using HRECG the time- and frequency-domain characteristics of the ECG signal were analyzed. The time-domain analysis was performed at 40-250 Hz filter. The frequency-domain analysis was performed by fast Fourier transformation (FFT), a 120 ms segment starting 20 ms before the end of the QRS complex was analyzed. The altered frequency content was expressed as the ratio of frequency areas (area ratio, AR) 20-50 Hz/0-20Hz. The average level of noise was 0.56 microV in the first group, 0.62 microV in the second group of patients and 0.68 microV in the control group. RESULTS: Abnormalities in the time domain analysis (ventricular late potentials, VLP) were present in 2 (13.3%) of 15 patients during the anthracycline therapy in the first group and in 4 (8.8%) of 45 patients after completing therapy in the second group. No abnormalities in the time-domain analysis were detected in the control group. Using frequency domain analysis, abnormalities in AR20-50 Hz/0-20 Hz were found in 8 (53.3%) of 15 patients of the first group, and in 11 (24.4%) of 45 patients of the second group. Significant differences were observed in the frequency parameters of the ECG signal in patients of the first group in comparison to the control group (p = 0.0018) and also when comparing the patients of the second group and the control group (p = 0.045). CONCLUSION: The HRECG results in time- and frequency-domain analyses indicate to high incidence of HRECG abnormalities in patients examined both during and after the antracycline therapy in comparison to the control group. The prognostic use of the HRECG abnormalities must be established in a larger and longer study. (Fig. 4, Tab. 2, Ref. 43.) PMID- 8705328 TI - [Acetylcholinesterase-positive innervation of the thymus in old rats after ovariectomy]. AB - Surgical ovariectomy in aged rats entails regeneration of the involuted thymus. These results correlate with our study of acetylcholinesterase positive innervation of regenerated thymuses. We found out a marked autonomous innervation of the perivascular tissues, as well as that of the functional parenchyma of the thymus. We proved a significant increase in the density of ACHE-positive nerves in the thymus of aged rats after surgical ovariectomy. These results are considered to represent and indirect evidence of the fact that cholinergic nerves stimulate lymphopoiesis in the thymus. (Fig. 5, Ref. 9.) PMID- 8705329 TI - [The circadian cycle of endothelin-1 in healthy persons]. AB - The possible circadian rhythmicity of endothelin-1 was investigated. Radioimmunoassay method was used in 5 clinically healthy male subjects aged 24-32 years. Blood samples were collected at 4-hour intervals over a period of 24 hours. Data for ET-1 were fitted to a 24-hour cosine curve. Statistically significant curves were found in two subjects (maximum at 4 p.m. and minimum at 10 p.m.), one did not yield a statistically significant curve and in two subjects despite the fact that the curves were not presented. Nevertheless, the circadian rhythmicity in our study was not confirmed according to our results it could not be excluded. (Fig. 2, Tab. 2, Ref. 23.) PMID- 8705330 TI - [Problems and perspectives of wider use of saliva for diagnostic purposes]. AB - Saliva of individual salivary glands differs in appearance, density and particularly in the chemical composition. Generally, the composition of saliva is affected by the composition of blood plasma, salivary flow rate, hormonal activity, drug administration, smoking and other physiological and pathophysiological states of the organism. In spite of these facts, many of the components are permanently present in saliva (e.g. peptides, enzymes, hormones...) only their concentrations vary. In some special cases unusual constituents can be detected in the saliva as legal and illegal drugs, antibodies (HIV), and abnormal bacteria or viruses. When there is good correlation between the levels of constituents in saliva and blood plasma then the determination of the constituent level in saliva can be used for diagnostic and/or monitoring purposes. But the main advantage of saliva analysis resides in stress-free and harmless collection of saliva in comparison with blood withdrawing. However, the use of saliva for diagnostic purposes is still at its beginning. So far, only few such applications are known, but optimists believe that the saliva analysis has a very prospective chance to substitute or alternate the biochemical analysis of blood plasma due to the mentioned advantage and to attribute more information on the processes in the human body. (Fig. 1, Ref. 16.) PMID- 8705332 TI - Choosing to be a children's nurse. PMID- 8705331 TI - The effects of prematurity on long-term outcome. PMID- 8705333 TI - The role of the neuro-oncology liaison nurse. PMID- 8705334 TI - Maintaining a paediatric N.D.U. PMID- 8705335 TI - How to audit Christmas. PMID- 8705336 TI - Cerebral palsy. PMID- 8705337 TI - Campaigning for adolescent care. PMID- 8705338 TI - Allitt: lest we forget. PMID- 8705339 TI - Choosing to be a children's nurse. PMID- 8705340 TI - Issues in bereavement: there are no rules. PMID- 8705341 TI - Paediatric bereavement care. PMID- 8705342 TI - The making of an N.D.U.--gaining accreditation. PMID- 8705344 TI - In the front line of child bereavement. PMID- 8705343 TI - The use of recombinant human dornase alfa in CF. PMID- 8705345 TI - RSV and bronchiolitis. PMID- 8705346 TI - Allitt inquiry. Malevolent intervention. PMID- 8705347 TI - Peer support group for adolescents. PMID- 8705348 TI - Management of gastro-oesophageal reflux. PMID- 8705349 TI - Parental bonding in cleft lip and palate repair. PMID- 8705350 TI - The role of the cardiac liaison nurse. PMID- 8705351 TI - Childhood immunisation. PMID- 8705352 TI - Health authorities fail to get the message. PMID- 8705353 TI - The art of storytelling. PMID- 8705354 TI - Travel the world of networking. PMID- 8705355 TI - Building bridges. Interview by Alison Turnbull. PMID- 8705356 TI - One-to-one midwifery: planning & development. PMID- 8705357 TI - Midwives and the future of maternity services. PMID- 8705358 TI - Small group practices. Part 3: The mother's perspective. PMID- 8705359 TI - Unhappiness after birth. PMID- 8705360 TI - Maternal infections. Part 1: Toxoplasmosis. PMID- 8705361 TI - Birth and the moon. PMID- 8705362 TI - To push or not to push? AB - Women in the second stage of labour instinctively bear down for short periods of four to six seconds. Most of these spontaneous bearing down efforts are accompanied by the release of air, with several breaths being taken between each effort. Conventional pushing techniques with sustained breath holding compromise fetal well being by reducing maternal arterial pressure and the oxygenation of maternal blood. Adequate progress may be made in labour, with the delivery of a healthy baby, when the management encourages spontaneous maternal efforts. PMID- 8705363 TI - Grave mistakes. PMID- 8705365 TI - Key developments in HIV maternity care. PMID- 8705364 TI - Everyday miracle ... or process? PMID- 8705366 TI - Look after your back. PMID- 8705367 TI - One-to-one midwifery. PMID- 8705368 TI - Time limits for court actions. PMID- 8705369 TI - We know what's best for you. PMID- 8705370 TI - The birth of Natasha. PMID- 8705371 TI - [Fifty years of medical genetics. A tribute to Maurice Lamy on the hundredth anniversary of his birth]. AB - Before 1960, no disease gene had been mapped to human chromosome apart from the sex-linked characters which are carried by the X chromosome. The first assignments were inferred from the results of somatic cells hydridization and concerned enzymatic deficiencies and protein defects. They were followed by the data gained from the cytogenetics studies of microrearrangements, either deletions or translocations. The family and linkage studies began to be successfully undertaken following the discovery of polymorphic probes. The breakthrough came out with the availability of highly polymorphic microsatellites scanning quite evenly, the major part of the genome. Currently, more than 1,000 clinical disorders are mapped and compiled in the database GID/GENATLAS. They concern all chapters and extend to characters which are usually sporadic, such as malignant tumours and congenital malformations. The mapping endeavour already had great impact on our understanding of fundamental life processes and unveiled the extent of genetic heterogeneity of diseases. Its major consequences concern prenatal diagnosis. The applications tend to extend towards presymptomatic diagnosis, and screening of carriers, two procedures still controversial, which request due consideration of their inherent risks and side effects and cannot be undertaken without the informed consent of patients. Predictive testing for the detection of liabilities is still a subject of lively debate. Although the spectacular advances of mapping and its implications open great hopes for the prevention and even cure of disease, care has to be taken to their limits and risks and, in their approach, full consideration must be given the respect if human person. PMID- 8705372 TI - [Reflections and proposals on the organization for pre-hospitalization of emergencies]. PMID- 8705373 TI - [Alcohol and free radicals: from basic research to clinical prospects]. AB - An oxidative stress occurs in the liver of rats following various conditions of ethanol administration. The ethanol-inducible cytochrome P450 2E1 plays a key role in its generation, favoured itself by an increase in the "redox-active " fraction of intracellular non-heme iron. Administration of ethanol elicits the generation of the 1-hydroxyethyl radical, which has been identified in vivo. Its reactivity contributes to alcohol-induced immunological disturbances. Liver inflammatory and fibrotic disorders can be reproduced in rats by long-term ethanol administration associated with a high fat diet. The severity of these disorders is correlated to the intensity of the oxidative stress. Some conditions of ethanol administration to rats also elicit an oxidative stress in the myocardium and central nervous system. Through its inhibitory effect on glutamine synthetase activity and resulting excitotoxicity it may contribute to neuronal death and possibly to dependence on alcohol. Disorders related to an oxidative stress were also reported in the serum and erythrocytes as well as in liver biopsies from alcoholic individuals. Their detection may be useful to follow the evolution of alcoholic liver diseases. Supplementation with antioxidants such as vitamin E may be considered in the prevention of severe cellular disorders in individuals consuming large amounts of alcoholic beverages. An increase in free radical production is likely playing a role in the induction of severe cellular damage linked to repeated withdrawals occurring as a result of heavy and sporadic ethanol intake. PMID- 8705374 TI - [Hysterectomy. Indications, abuse, psychological impact]. AB - Statistical surveys have established that, in France, hysterectomy was carried out too frequently: 50 to 70,000 hysterectomies (perhaps only 40,000) are carried out each year in our country. This fact, together with the deleterious psychological effects of this operation, have urged the authors to express some remarks on the current practice of hysterectomy in France. The currently approved indications are the following: invasive cancer, benign lesions with intractable disabling symptoms and technical requirements (e.g. cure of prolapse in the elderly). Bleeding due to fibromyomas should be managed with conservative means as much as possible. Infections are within the scope of medical treatment. When an hysterectomy is indicated, the vaginal route should be preferred whenever possible, due the lesser impact on the body image and personality. The adverse psychological effects of hysterectomy are due to several factors: fear of the operation, pain, possible complications, breach of the femininity, alteration of the body image, fear of menopause and ageing. These effects could be reduced by an accurate information of the patient on the actual consequences of the operation, which are far from considerable, specially if the hormonal secretion is preserved. Also, a certain period of time between the decision and the operation might help to further reduce this psychological impact. PMID- 8705375 TI - [Eulogy of Henri Baylon (1913-1994)]. PMID- 8705376 TI - [Bovine spongiform encephalopathy and Creutzfeldt-Jakob disease. Current questions]. AB - The occurrence of Creutzfeldt-Jakob disease (CJD) in two teenagers and four dairy farmers in the United-Kingdom has focused attention on the possibility of a causal link between bovine spongiform encephalopathy (BSE) and Creutzfeldt-Jakob disease. The BSE epidemic was initiated by the inclusion in the cattle feed of the ruminant-derived protein in 1981/1982. A feed ban of the ruminant-derived protein in 1988 prevented further transmission of the infective agent to cattle by this route. However cases in animals born some years after the feed ban suggest that there has been some continued leakage of BSE infected material into animal feed (and perhaps maternal transmission?). To protect public and animal health, measures have been taken in the United Kingdom, especially prohibition of specified bovine offals in 1989 (extending this ban to thymus and intestine of calves slaughtered for human consumption in 1994). These measures are considered as minimizing the risk of BSE infection. Although the BSE linked risk factors are probably unique to the United Kingdom, it seems necessary to have the same regulations for the calves imported from this country. More, it is crucial that the ongoing surveillance programme of CJD in Europe is sustained. PMID- 8705377 TI - [Infection of the mouse by Theiler's virus; a model for the study of demyelinating diseases of the central nervous system]. AB - The infection of mouse by Theiler's virus is a model for the study of persistent infections of the central nervous system accompanied with primary demyelination. The pathogenesis of this disease has been studied at the molecular level. The viral capsid controls the migration of the virus from the neurons to the glial cells of the white matter. A region of the capsid, which is responsible for this control, has been identified. The susceptibility of mice to this persistent infection is determined by at least three genes, one of which, H-2D, has been identified. Two other genes have been mapped; one linked to the gene coding for gamma interferon, the other one to the gene coding for the myelin basic protein. The role of cytotoxic T lymphocytes in the resistance to persistent infection has been demonstrated. Taken together, these results illustrate how mutations in a ubiquitous virus may cause, in individuals which are genetically susceptible, a persistent infection and a chronic inflammatory disease. Similar situations could exist in man. PMID- 8705378 TI - [Role of plastic surgery in the treatment of breast cancer]. AB - The surgical treatment of breast cancer has changed dramatically since the last twenty years. Radical and super-radical mastectomy are almost no longer indicated except in certain cases of local recurrences. Breast saving procedures are the best indication for small tumors. And the psychological impact of the different treatments including radiotherapy and chemotherapy is considered as an important issue. Therefore, plastic surgeons are more and more frequently called upon to restore the breast appearance using some technique of breast reconstruction or to improve the bad results of a conservative treatment. Moreover, a good knowledge of the plastic surgery techniques allows a better exploration of the breast and large resections in case of fibrocystic diseases with extensive foci of microcalcifications. Such extensive explorations of the breast are more frequently indicated with the development of screening campaigns. When the mastectomy remains the best indication, it is mandatory to propose an immediate breast reconstruction. Insertion of a silicone implant is discussed in several countries, especially in France, although more and more studies are showing the inocuity of such material. A long experience at the Gustave-Roussy Institute of such material used for breast reconstruction showed us the inocuity of long term silicone exposure in patients treated for breast cancer. The conservative surgery should remove the tumor with a free margin of 1 to 2 or even 3 cm and the defect left is sometimes large enough to justify some local glandular mammoplasty in order to prevent from final mammary distorsion or local visible defect. Partial reconstructions could sometimes require more sophisticated techniques including musculo-cutaneous flaps or prosthesis implantation. Finally the integration of the plastic surgery in the surgical protocols of breast cancer treatment, requires a close collaboration between the plastic and the cancer teams. In certain cases, a special surgical training provides the surgeon with a double competence in oncology and in plastic surgery. With such breast surgeon competent both in oncology and plastic surgery, a patient can ask for an aesthetic operation without missing a complete cancer screening as well as she can be treated for a cancer with the best chance to obtain the best cosmetic result. PMID- 8705379 TI - [Induced tissue transformation and heart surgery]. AB - This article reports the research which led to the use of animal connective tissues in the construction of valvular prostheses and those which led to the use of electrically stimulated skeletal muscle for cardiac assistance. Although, very different at first glance these research have in common the transformation of biological tissues by physical or chemical means to adapt them to a new function. 1) Once implanted in a different species, animal connective tissues are destroyed by immunological reactions and collagen degeneration. These lesions can be prevented by both maskage of the antigenic groups and intermolecular crosslinking using Glutaraldehyde. The durability of such chemically treated tissues is based upon the stability of the biological material (concept of bioprosthesis) and not upon cell survival or tissue regeneration by host cell ingrowth (concept of graft). The valvular bioprostheses made from Glutaraldehyde treated pericardial tissue, keep after this treatment their advantage of biological tissues: they are not thrombogenic and do not require anticoagulation contrary to mechanical valves. Although they have a limited durability up to 10 to 15 years due to tissue calcification, they represent 40% of the valvular prostheses used in clinical practice today. 2) The clinical use of electrostimulated skeletal muscle has been delayed for a long time because of fatigue lesions. An original protocol of progressive sequential stimulation prior to the use of muscle prevents fatigue by the transformation of type I fatigable myosin into type II non fatigable myosin. The conditionned muscle i.e.: the latissimus dorsi, is then wrapped around the ventricles to either reinforce cardiac contraction or to replace a portion of the heart. In the past 10 years, this new operation of "dynamic cardiomyoplasty", has been performed in 84 patients suffering from the end stage heart failure in our institution and in over 500 patients throughout the world with significant functional improvement. PMID- 8705381 TI - [Medical assistance for procreation. Current medical problems]. PMID- 8705380 TI - [Wearable mechanical ventricular support]. AB - Progressive technological developments have permitted clinical use of the wearable Baxter-Novacor left ventricular assist system. The system allows total bypass of the left ventricular function, and recovery of an adequate circulation in patients about in cardiogenic shock. Since the first clinical use, in March 1993, six patients about to die have been supported with a cardiac transplantation. This experience suggests that an acceptable answer to the problem of organ shortage is non available. PMID- 8705382 TI - [The discovery of insulin]. AB - When a medical problem is intensively studied by many teams in the world, it is frequent to see the solution found simultaneously in different countries. However that was not exactly the case concerning the extraction of a potent insulin able to cure Diabetes Mellitus. It seems necessary, seventy five years later, when passions are quenched, to reconsider the chronology of the history and put Paolesco but also Collip at the right places much before Banting and Best to whom, by a curious misinterpretation of facts, was attributed the priority of this fundamental discovery. PMID- 8705383 TI - [Clinical, morphologic and immunophenotypic data based on 10 cases of canine muco cutaneous epidermotropic T-lymphoma (analogous to Mycosis Funcgoide). Important of an animal model of spontaneous pathology]. AB - Our serie of ten canine cutaneous epitheliotropic T-cell lymphomas (CTCL), is found in old dogs, belonging mainly to the Boxer breed. Site on the mucous membranes (especially buccal), the muco-cutaneous junctions, their clinical expression is polymorphous. Lesions, follow on one after another (erythema, plaques, nodules) and are diversely associated in a given animal, the borders between the different stages often being difficult to establish. Adenopathies noted at the time of the diagnosis or during the course of the condition are accompanied by an involvement of the blood and organs (analogous to Sezary's disease). The progression of the disease can be very rapid in the buccal forms, which are generally aggressive, and in cases of violent, uncontrollable pruritus, which may be disturbing for the owner (with requests for euthanasia). The neoplastic infiltrate is constituted of small lymphocytes with hyperchromatic, convoluted nuclei (incipient stages), then large cells with a "histiocytic" appearance for the nodules. Epitheliotropism, which is maximal for the infiltrated plaque stage, shows up either in the form of a flux of totally epitheliotropic isolated cells (Ketron-Goodman type) or in that of Pautrier abscess-like collections. THe veterinary literature is in agreement that the CTCL cell expresses CD3, but two recent studies are in contradiction as regards its membership of helper or cytotoxic/suppressor populations. For our 10 cases, all the cells of lymphocytic morphology were, without exception, CD3+ and CD45+, irrespective of their situation within the epithelium or the chorion. The CD3+ cells in the epithelium were systematically CD8+, CD4- (confirming P.F. Moore's observations), expressing CD5 in a variable way, and, mostly, the Ki-67 nuclear proliferation Ag. The CD3+ cells of the chorion were exclusively, or mainly, CD8+, and occasionally CD4+. They expressed CD5 in a variable way, and, for a minority, the Ki-67 nuclear proliferation Ag. On the pathogenic level, it may be suggested that a T clone, CD8+, undergoes the "homing" phenomenon within the epithelium, enters the cell cycle, then manifests a tropism towards the chorion, which it infiltrates. Despite some particularities, which may be clinical (serious mucous attacks), cytological (the "histiocytic" appearance of the nodule cells) or immunophenotypic (expression of CD8, similar to what is observed in man in a considerable number of Pagetoid reticulosis), CTCL constitutes an interesting model of spontaneous pathology, and could prove useful in: - identifying various etiological factors (given that the dog, as a close commensal of man, is subject to the same environmental factors). PMID- 8705384 TI - [Responsibilities and generic drugs]. AB - The existence of generics is essentially due to the competition between pharmaceutical specialties by the prices at the end of patent protection. It could be useful in the countries where the prices are very high. In France, the consumption of drugs is high in volume and the average of prices of pharmaceuticals is low. So the generics is not able to really solve the problem of pharmaceuticals' consumption. Moreover one must be vigilant from healthcare point of view for the quality of production. The consequences at the level of pharmaceutical researches could be significant. At last, the eventual right substitution can threaten the freedom of prescription. PMID- 8705385 TI - [Angiotensin IV, a new component of the renin-angiotensin system, which acts on kidney cells]. AB - Angiotensin IV (Ang IV), the hexapeptide obtained from angiotensin II (Ang II) by deletion of the first two N terminal amino acids, possesses specific receptors in various tissues. Our aim was to search for such receptors in two types of renal cells, rat mesangial cells and principal cells of the human collecting duct. [125I]-Ang IV specifically bound to mesangial cell surface and to membranes prepared from the principal cells. In both cases, affinity was approximately 5 nmol/L and receptor density was close to 1000 fmol/mg protein. The order of potency of different competitors was as follows: Ang IV > Ang III > Ang II > Ang II (4-8) > Ang II (1-7). Binding sites were distinct from those of Ang II since type 1 or type 2 Ang II receptor nonpeptide antagonists produced no displacement. Reciprocally, Ang IV did not displace Ang II from its binding sites. Ang IV inhibited the vasoconstrictor effect of Ang II on rat mesangial cells and increases cyclic AMP production in principal cells, but only when it had been previously stimulated. Taken together, these results demonstrate that the glomerulus and the collecting duct represent target sites for Ang IV and suggest that Ang IV could influence the renal functions. PMID- 8705386 TI - Effect of hemorrhagic shock and resuscitation upon hepatic phagocytic clearance and killing of circulating microorganisms. AB - We evaluated the effects of hemorrhagic shock/resuscitation (S/R) on hepatic reticuloendothelial system function, using an in vivo assay that discriminantly quantitates its two essential components, phagocytic clearance and phagocytic killing of double-labeled Escherichia coli injected intravenously. Rats were subjected to hemorrhagic shock at mean arterial pressure at 50 +/- 5 torr for 2 h, resuscitated, and survived. Hepatic phagocytic clearance was significantly decreased immediately following and 6 h after S/R, compared with sham-shocked rats, but recovered to normal levels after 24 h. Although hepatic killing efficiency was increased initially, and transiently depressed 6 h later, it was strikingly upregulated after 24 h. As a consequence, net hepatic killing was transiently suppressed at 0 and 6 h, but upregulated after 24 h. Pre-treatment with proinflammatory agonists, including endotoxin, IFN-gamma, or IFN-gamma + TNF alpha enhanced both hepatic killing efficiency and net hepatic killing. These observations suggest that although hepatic killing function is initially impaired after the onset of S/R, it is strikingly upregulated 24 h later, simulating both the initial immunosuppression, and the later hyperinflammatory response to systemic S/R that could lead to multiple organ dysfunction syndrome. PMID- 8705387 TI - Alterations in tumor necrosis factor-alpha expression by hepatic macrophages following acute cholestatic liver injury. AB - The liver is unique for its large resident macrophage (HM phi) population as a potential source of immunoregulatory cytokines. The present study was designed to determine HM phi function in a rat model of cholestasis (CBDL). Northern blot analysis of TNF-alpha mRNA showed a profound difference in the dose response to bacterial lipopolysaccharide (LPS) between sham and CBDL HM phi. Sham HM phi demonstrated an 8-fold difference in induction of TNF-alpha mRNA versus CBDL HM phi. TNF-alpha secretion, determined by enzyme-linked immunosorbent assay, was significantly higher from LPS-activated sham HM phi versus the same cells activated with Gram-positive bacterial peptidoglycan while CBDL HM phi were more responsive to peptidoglycan than to LPS. These results demonstrate stimulus- and response-specific functional alterations in the HM phi population during acute cholestatic injury. We speculate that these functional alterations are phenotypically induced in acute liver injury resulting in responses that are not characteristic of normal HM phi. PMID- 8705388 TI - A selective inhibitor of inducible in nitric oxide synthase prolongs survival in a rat model of bacterial peritonitis: comparison with two nonselective strategies. AB - We evaluated the effects on survival of three different strategies for blocking the actions of nitric oxide (NO) during Gram-negative sepsis in rats. Male Sprague-Dawley rats underwent placement of a jugular vein catheter and i.p. implantation of a gelatin capsule containing a paste (.11 +/- 0.1 g final weight) consisting of sterile rat feces mixed with a suspension (.2 mL) of viable Escherichia coli (strain sm 18; 5.7 x 10(5) colony-forming units) in saline. Beginning at T = 6h, all animals received i.v. ampicillin (85 mg/kg every 12 h) until death or the administration of five doses. At the same time points, pairs of animals received an i.v. dose of either an experimental treatment agent or an appropriate control substance. The following experimental regimens were tested: 5 mg/kg per dose of S-methylisothiourea sulfate (SMT), a selective inhibitor of the inducible isoform of nitric oxide synthase (NOS); 10 mg/kg per dose or 25 mg/kg per dose of NG-nitro-L-arginine methyl ester (L-NAME), an inhibitor of the inducible and constitutive isoforms of NOS; 200 mg/kg per dose of cross-linked human hemoglobin (HGB), an NO scavenger. SMT significantly prolonged survival in septic rats, although cumulative survival at T = 168 h was approximately equivalent in SMT- or saline-treated animals. In contrast, HGB and the higher dose of L-NAME significantly shortened survival times. At T = 20 h, arterial PO2 was significantly lower in rats treated with HGB as compared to time-matched controls. We conclude that SMT, a compound with reported activity as a selective inhibitor of the inducible isoform of NOS, prolongs survival in a rat model of antibiotic-treated Gram-negative sepsis. PMID- 8705389 TI - Role of nitric oxide in sepsis-induced hyporeactivity in isolated rat lungs. AB - The aim of the present study was to test the hypothesis that pulmonary microvascular reactivity is depressed in sepsis and that inducible nitric oxide synthase (iNOS) contributes to the vascular hyporeactivity. Rats were made septic by cecal ligation and puncture. After 16 h, pulmonary vascular reactivity was evaluated by measurement of perfusion pressures while the vasculature was challenged with angiotensin II and KCl. The results showed that vascular reactivity was significantly depressed in lungs from septic rats in comparison to sham-operated controls. Pretreatment with the nitric oxide synthase inhibitor NG nitro-L-arginine methyl ester (L-NAME, 100 microM) restored the depressed vasoreactivity while the nitric oxide (NO) synthase substrate L-arginine (1 mM) reversed the contraction-restoring effect of L-NAME. NO production in lungs from septic rats increased about 4-fold in comparison to sham-operated controls. iNOS protein was expressed in lung tissues, mainly the resistance vessels, from septic rats but not from sham-operated controls. Reverse transcription and polymerase chain reaction also showed a strong induction of iNOS mRNA in lung tissues from septic rats. These results suggest that increased iNOS expression and NO production may contribute to depressed pulmonary vascular reactivity in sepsis. PMID- 8705390 TI - Hypertonic saline in stabilized hyperdynamic sepsis. AB - Hypertonic saline with or without colloidal solution has been successfully used for treating hemorrhagic shock in animal experiments and clinical studies. Due to its various effects at systemic, organ, and microcirculatory levels, the substance appears to be a promising candidate for improving tissue oxygenation in sepsis. We therefore investigated the hypothesis that infusion of hypertonic saline would further improve O2 delivery, O2 extraction, and O2 uptake in hyperdynamic septic shock patients already stabilized by adequate volume and catecholamine infusion. Twenty-one patients received 2-4 mL/kg body weight of hypertonic saline in hydroxyethyl starch within 15 min. This hypertonic saline infusion caused a rapid significant increase in O2 delivery by 14% but only a marginal increase in O2 consumption (7% by cardiovascular Fick [p < .05], 4% by respiratory gases [n.s.]). Hypertonic saline increased the already elevated cardiac output by 24%. The pulmonary capillary wedge pressure increased from 14 +/- 3 to 23 +/ 3 mmHg and pulmonary shunt fraction increased 15%, but arterial PO2 did not fall. Except for the increase in pulmonary capillary wedge pressure, none of the cardiovascular changes lasted longer than 60 min. Plasma sodium levels increased from 138 +/- 25 to 163 +/- 38 mmol/L and normalized within 24 h. In these hyperdynamic septic patients, hypertonic saline infusion produced a transient increase in circulation, but no evidence of a substantial increase in O2 consumption. Either there was no significant O2 debt due to the already elevated O2 delivery levels at baseline (700 mL/min/m2) or the global O2 measurements we used were not able to detect discrete regional hypoxia. PMID- 8705391 TI - The fall of cardiac output in endotoxemic rats cannot explain all changes in organ blood flow: a comparison between endotoxin and low venous return shock. AB - During endotoxin shock mean arterial pressure (MAP) and cardiac output (CO) fall, and the latter is redistributed. To evaluate whether these changes are solely caused by the low output, or are also based on endotoxin itself, we compared regional hemodynamic changes during endotoxemia with those in a nonendotoxemic state of decreased CO in anesthetized rats. In group E (n = 10) endotoxin Escherichia coli O127:B8 (8 mg.kg-1) was infused from t = 0 till t = 60 min. In group B (n = 10) the same decrease of CO and MAP was obtained as in group E by inflating a balloon in the inferior caval vein, distal to the renal veins, from t = 0 till t = 60 min. We measured MAP, CO (thermodilution), central venous pressure, heart rate, organ blood flow, and redistribution of CO (microspheres), arterial lactate and glucose, and hematocrit. MAP and CO decreased (p < .05) in both groups (by 30 and 50%, respectively at t = 60). Heart rate, hematocrit, arterial lactate, and arterial glucose were significantly higher (p < .05) in group E (by 17, 12, 180, and 55%, respectively). Blood flow to most organs had similarly decreased in both groups. The decreased intestinal blood flow lead to macroscopic damage only in group E. Blood flows (absolute or as percentage of CO) to heart, hepatic artery, and diaphragm, however, had significantly increased in group E while blood flows to skin, skeletal muscle, and stomach had decreased more in group E. Except for the heart these differences could be explained by increased work load (detoxification: liver; hyperventilation: diaphragm, muscle) and thus to a more pronounced redistribution at the expense of skin and muscle blood flow. Regional hemodynamic changes during endotoxemia thus could largely be attributed to decrease of CO and redistribution of the circulating blood volume. In the heart, endotoxin seemed to exert effects independent of the hypodynamic state. This was also true for the intestinal damage and the rise in hematocrit and arterial lactate. PMID- 8705392 TI - The effects of diaspirin cross-linked hemoglobin in sepsis. AB - We tested the hypothesis that diaspirin cross-linked hemoglobin (DCLHb; Baxter Healthcare Corp.) would improve blood pressure, organ perfusion, and mortality during sepsis. Rats were catheterized to assess general hemodynamics (protocol 1) or regional blood flow (protocol 2). Sepsis was induced by intraperitoneal introduction of a cecal slurry (100 mg/kg). In protocol 1, rats received either 100 or 250 mg/kg DCLHb, or albumin at 1, 2, or 4 h after sepsis induction. Hemodynamics were recorded at these times and daily for 72 h. DCLHb increased blood pressure, prevented 72 h leukocytosis, and reduced mortality, but the timing of DCLHb administration was crucial. In protocol 2 only moribund septic animals received 100 mg/kg DCLHb or iso-oncotic albumin i.v. Hemodynamics and regional organ blood flows were measured at baseline, immediately before and after treatment, and at 24 h. DCLHb immediately increased blood pressure with no changes in cardiac output, heart rate, or regional perfusion. DCLHb increased regional perfusion to vital areas at 24 h (compared to albumin group). Distribution of cardiac output in albumin-treated rats was significantly skewed toward skeletal muscle at a time when cardiac output was significantly lower as compared with DCLHb treated animals. In conclusion, DCLHb safely elicited a pressor response, and improved regional perfusion to selected tissues. However, DCLHb benefits were best obtained when given within a specific time frame. PMID- 8705393 TI - Efficacy of convective removal of plasma mediators of endotoxic shock by continuous veno-venous hemofiltration. AB - Continuous veno-venous hemofiltration (CVVH) has been reported to provide beneficial effects during endotoxic shock. This experiment was designed to determine if selective removal of plasma mediators occurs during CVVH and if plasma concentrations of these mediators are reduced. A swine endotoxic-shock model with three groups was used (lipopolysaccharide (LPS) only (n = 6); LPS followed by CVVH (n = 6); and LPS followed by sham CVVH (n = 4). Plasma and filtrate samples were collected at frequent intervals for 5 h. Lactic acid (LA), eicosanoids [prostacyclin (6-keto PGF1 alpha), thromboxane (TxB2), and prostaglandin E2 (PGE2)] and tumor necrosis factor (TNF) were measured in plasma and filtrate. Plasma concentrations of 6-keto PGF1 alpha, TxB2, TNF, and LA were not significantly different in any group. LA, PGE2, 6-keto PGF1 alpha, and TXB2 concentrations were similar in filtrate and plasma. TNF did not move across the membrane into the filtrate, CVVH, as used in this experiment, did not significantly reduce plasma concentrations of any of these mediators. PMID- 8705394 TI - Trauma-induced suppression of antigen presentation and expression of major histocompatibility class II antigen complex in leukocytes. AB - The immune response to trauma, shock, and/or sepsis appears to exhibit a bimodal response, in which there is an early exaggerated inflammatory response, giving way over time to a state of hyporesponsiveness or immune dysfunction. This state of immune dysfunction is frequently associated with increased infectious complications and/or mortality, seen following shock or trauma. In this article, we present an overview of some of those changes that have been seen with respect to the process of major histocompatibility class II (MHC class II) antigen presentation by macrophage, a key component of the overall host immune response to foreign bacterial and/or fungal pathogens encountered following shock/trauma (with a particular emphasis on hemorrhagic shock as a component of traumatic shock). With respect to the overall process of antigen presentation, defects (dysfunction) are evident not only in models of shock and sepsis, but also in traumatized patients. Studies of the capacity of a monocyte's/macrophage's ability to present antigen indicate that defects can be detected, not only in those steps involved in antigenic processing, but also in MHC class II molecule expression and accessory molecule function (or its inhibition) following shock. Those changes in the macrophage's capacity to process antigen seen during the first 24 h after hemorrhagic shock appear to be associated with the cell's metabolic response to regional hypoxia and/or the shift to proinflammatory mediator release (tumor necrosis factor, interleukin [IL]-1, IL-6, etc.). This initial acute response to shock appears to act as the nidus for chronic anti inflammatory mediator release (prostaglandin E2, transforming growth factor-beta, IL-10, IL-4, nitric oxide, etc.), which may mediate the sustained depression of the antigen-presenting cell's function. PMID- 8705395 TI - A phase I safety and pharmacokinetic study of a recombinant amino terminal fragment of bactericidal/permeability-increasing protein in healthy male volunteers. AB - A phase I pharmacokinetic and safety clinical trial of rBPI23, a recombinant amino terminal fragment of bactericidal/permeability-increasing protein, was conducted in healthy male volunteers. rBPI23 was administered as a 5 or 30 min infusion at doses of .1 to 1 mg/kg. The pharmacokinetics of rBPI23 in human subjects were described by a bi-exponential disposition function with evidence of concentration-dependent kinetics. The alpha half-life increased significantly with increasing dose, from 4-5 min at .1 mg/kg to 7-8 min at 1 mg/kg. The beta half-life varied between 18 and 29 min regardless of dose and the clearance varied from 5 to 10 mL/min/kg. Very little, if any, of the administered rBPI23 was excreted intact in the urine. Electrocardiograms, ionized calcium concentration, prothrombin and partial prothrombin times, hematologic parameters, and blood chemistries remained normal. Furthermore, no antibody response to rBPI23 was observed in any of the subjects. PMID- 8705396 TI - Altered oxygen tension modulates cytokine-induced signal transduction in polymorphonuclear leukocytes: regulation of the GPLD pathway. AB - We investigated the effect of alterations in buffer oxygen tensions from normoxia (PO2 = 180-200 mm/Hg) to hypoxia (PO2 < 30 mm/HG) and then reoxygenation (PO2 > 140 mmHg) on the GPLD-pathway by measuring phosphatidylethanol formation in the presence of ethanol and subsequent NADPH oxidase activation and O2-production in polymorphonuclear leukocytes (PMN). Experiments were performed with PMN stimulated with either interleukin (IL)-8, tumor necrosis factor (TNF)-alpha, or IL-1 beta in the presence or absence of fibronectin. Hypoxia exerted a downregulating effect on this pathway and reoxygenation restored GPLD activation to levels seen during normoxia; however, supraphysiological concentrations of cytokines were able to reverse this pattern. Changes in GPLD activation correlated best with changes in O2-production during the hypoxia to hypoxia/reoxygenation transition induced by TNF-alpha-Fn and IL-1 beta +/- Fn. Thus, changes in oxygen tension can directly modulate the extent of the PMN response to stimulation by IL-8, TNF-alpha, or IL-1 beta, and activation of the GPLD-pathway appears to be highly sensitive to hypoxia and hypoxia/reoxygenation. PMID- 8705397 TI - An expert diagnostic system based on neural networks and image analysis techniques in the field of automated cytogenetics. AB - In this study, we introduce an expert system for intelligent chromosome recognition and classification based on artificial neural networks (ANN) and features obtained by automated image analysis techniques. A microscope equipped with a CCTV camera, integrated with an IBM-PC compatible computer environment including a frame grabber, is used for image data acquisition. Features of the chromosomes are obtained directly from the digital chromosome images. Two new algorithms for automated object detection and object skeletonizing constitute the basis of the feature extraction phase which constructs the components of the input vector to the ANN part of the system. This first version of our intelligent diagnostic system uses a trained unsupervised neural network structure and an original rule-based classification algorithm to find a karyotyped form of randomly distributed chromosomes over a complete metaphase. We investigate the effects of network parameters on the classification performance and discuss the adaptability and flexibility of the neural system in order to reach a structure giving an output including information about both structural and numerical abnormalities. Moreover, the classification performances of neural and rule-based system are compared for each class of chromosome. PMID- 8705399 TI - A device for the measurement of malleoli diastasis. AB - Assessment of the degree of laxity of the distal tibio-fibular joint ligaments is of great importance in the treatment of some ankle pathologies resulting from traumas and when evaluating the effectiveness of re-constructive operations. This can be done by measuring the variation in the intermalleolar distance while loading and flexing the joint in a known way. So far, only X-ray techniques have been applied for this purpose, but they are invasive and do not allow for analysis under dynamic conditions. We have developed a wearable strain gauge device to be positioned directly over the malleoli for measurement of the intermalleolar distance variation. It has been tested on healthy and injured subjects, in both static and dynamic conditions. Data obtained in the static tests are in agreement with those found in literature. Dynamic analysis allowed for correlation of malleoli diastasis (MD) with the different phases of gait and different injured states. PMID- 8705398 TI - In vitro testing of a left ventricular assist device. Study of the effect of its control strategy on energetic relationships inside the left ventricle. AB - In this study an original left ventricular assist device is tested on an open loop modular physical circuit reproducing Starling's law of the heart to set an optimal control strategy for heart recovery. It is assumed that the goals of the assistance are reduction of oxygen consumption, external work and improvement of cardiac mechanical efficiency. The assistance is evaluated by the position of a working point on the characteristic surfaces of the ventricle defined by peripheral resistance, atrial pressure and selected variables pertaining to energy (pressure-volume area, external work and cardiac mechanical efficiency). In this frame an optimal assistance for heart recovery is a compromise among different requirements corresponding to a restricted set of control parameters values: driving pressure PZB = 35 kPa and timing values T1 and T2 (systole beginning and systole ending in relation to QRS complex and cardiac cycle duration T) T1 = 0.55.T and T2 = 0.73.T. PMID- 8705400 TI - Optoelectronic techniques for patient repositioning in radiotherapy. AB - In modern radiotherapy practice, patient mispositionings and movements at therapy units are considered one of the major source of error in the irradiation and, consequently, a decisive factor in relapse rate and in radiation damages occurrence. Movement analysis system ELITE has been used to carry out a new conceived control on patient repositioning procedure and to implement an automatic correction of the detected position errors. Two marker disposition models have been defined for the supine position with reliable and easily identifiable body landmarks. Analytical comparison between current and reference 3D co-ordinates of these points has constituted the core of the whole method. The detection of patient's breathing phases has allowed to synchronize the co ordinates calculation to patient's FRC, increasing accuracy and reliability of the comparison. Results point out the relative accuracy of the commonly adopted optical laser repositioning systems and confirm the significant role that ELITE can have in radiotherapy, in order to reach a suitable tradeoff between a quick irradiation set-up and a clinically correct patient irradiation. PMID- 8705402 TI - A method for predicting the utilization of medical technology in neonatal care. AB - The utilization of 12 neonatal intensive care technologies was recorded in 193 patients. An index of utilization (Pu) was adopted to classify these technologies into three sub-groups corresponding to low (Pu < 15%), medium (15% < or = Pu < or = 85%), and high (Pu > 85%) intensity of use. No models were developed for technologies with high intensity of use (intravenous hydration, warming, micro haematocrit, heart rate monitoring). Multiple linear regression was used to predict the utilization of technologies with medium intensity of use (gasometry, oxygen hood, CPAP, mechanical ventilation), and multiple logistic regression was used with the same purpose for technologies with low intensity of use (exchange transfusion, bicarbonate, adrenaline, echocardiography). Both methods yielded significant models (P < 0.05) which can be used to improve planning and management of technology in neonatal intensive care units. PMID- 8705401 TI - Laparoscopic and thoracoscopic surgery in infancy and childhood, the Munster/Gent experience. AB - Laparoscopic and thoracoscopic surgery have had a tremendous impact on adult surgery, but are still rarely used in children. In the past 3 years 168 children have been treated endoscopically in the three cooperating institutions. The course of all patients was documented prospectively in order to determine the value and prove the safety of endoscopic surgery in children. The operations performed endoscopically comprised appendectomy (n = 39), diagnostic procedures (24), adhesiolysis (n = 22), cholecystectomy (n = 13), bowel resection (n = 8), pyloromyotomy (n = 6), ovarial resection (n = 6), hiatus hernia repair (n = 22), splenectomy (n = 6), first stage of Fowler-Stephens operation for kryptorchidism (n = 3) and interruption of persistent botallic ducts (n = 6) in preterm infants. Apart from four wound infections (2.4%) following perforated appendicitis, one of which required laparotomy for perityphlitic abscess there were no serious complications. No patient died, and overall morbidity related to the procedure was 1.8% (i.e. incidence of uneventful minor intraoperative complications). Furthermore endoscopic surgery provides a better diagnostic survey of the abdomen or thorax so that missing of secondary pathology is unlikely. Pain and reflectory impairment of bowel-function or respiration were diminished, the aesthetic results were excellent and hospitalization could be reduced to a minimum. It is concluded that laparoscopic and thoracoscopic surgery are safe and reliable procedures for children and yield very encouraging results. PMID- 8705403 TI - The CARDIO-LOGOS system for ECG training and diagnosis. AB - A new approach in ECG training is presented. The overall approach is based on a combination of the "page-turning architecture", the "reference model" and AI techniques. A thorough analysis of the training requirements in this field has been carried out; the results determined the educational scenarios and the associated evaluation sessions. The system (CARDIO-LOGOS) is intended to be used mainly by internal medicine physicians and general practitioners, as well as by medical students. The layered structure of the whole CBT application together with the advanced learning strategies and the interactive multimedia technology offer a flexible environment that encourages experimentation and supports individualised training for a wide variety of users. PMID- 8705404 TI - Obstetrics and gynecology ultrasound laboratory accreditation in the USA. PMID- 8705405 TI - Prenatal detection of lethal pulmonary hypoplasia. PMID- 8705406 TI - Sonography of pregnancies with first-trimester bleeding and a viable embryo: a study of prognostic indicators by logistic regression analysis. AB - The objective of our study was to investigate the relationship between sonographic findings and the occurrence of abortion in pregnancies complicated by first-trimester bleeding in which fetal cardiac activity was documented upon admission. A prospective study of transvaginal sonography was performed in 270 pregnant patients with bleeding between 5 and 12 weeks' gestation. The study group included 149 cases in which a singleton fetus with cardiac activity was initially documented. The outcome variable was pregnancy loss prior to 20 weeks. The influence of sonographic findings on admission was studied by univariate analysis and logistic regression. The prevalence of abortion was 23/149 (15%). A significant relationship (p < 0.05) was found between the occurrence of abortion and the following: fetal bradycardia (heart rate less than -1.2 SD from the mean), a discrepancy between the diameter of the gestational sac and crown-rump length less than -0.5 SD from the mean, and a discrepancy between menstrual and sonographic age of more than 1 week. According to the logistic regression equation that was obtained, the probability of abortion in first-trimester bleeding with documented fetal cardiac activity upon admission varied between a minimum of 6% when none of the above risk factors were present and a maximum of 84% when all were present. The presence of any of the above factors identified 84% of all subsequent abortions. PMID- 8705407 TI - The prevalence of non-viable pregnancy at 10-13 weeks of gestation. AB - In an ultrasound screening study at 10-13 weeks of gestation involving 17,870 women, the prevalence of early pregnancy failure was 2.8% (501 cases), including 313 (62.5%) missed abortions and 188 (37.5%) anembryonic pregnancies. Lower gestation and higher maternal age were associated with a higher prevalence (chi 2 = 143.5; p < 0.001 and chi 2 = 53.3; p < 0.0001, respectively). The prevalence was higher in women with a history of vaginal bleeding (chi 2 = 141.5; p < 0.0001), but there was no significant association with previous pregnancy losses (chi 2 = 2.8), parity (chi 2 = 0.6) or cigarette smoking (chi 2 = 0.0). Recent evidence suggests that the most effective method of screening for chromosomal abnormalities is measurement of fetal nuchal translucency thickness at 10-13 weeks, and therefore ultrasound examination at this gestation is likely to become universally available. As shown in this study, an additional advantage of such a scan is the diagnosis of early pregnancy failure, which will be found in about 3% of patients examined. Elective evacuation of retained products of conception is likely to be more cost effective and potentially safer than emergency surgery in a patient presenting during miscarriage. PMID- 8705408 TI - Renal duplication anomalies in the fetus: clues for prenatal diagnosis. AB - Duplex kidneys are one of the most common major congenital abnormalities of the urinary tract. The antenatal diagnosis of duplex kidney and its associated ureterocele is infrequent. We report on our experience with the prenatal diagnosis of duplex kidneys in seven fetuses over the past 24 months. In all fetuses, the sagittal length of the duplex kidney was above the 95th centile for gestational age. A 'cyst-like' structure in the upper pole of the duplex kidney and a ureterocele in the urinary bladder were present in all of the seven fetuses. An ipsilateral dilated ureter was seen in six of seven fetuses. Postnatal confirmation of renal duplication anomalies was obtained in all neonates. Increased familiarity of the prenatal sonographer with duplex kidney will allow for its antenatal diagnosis and thus early postnatal treatment. PMID- 8705409 TI - Ultrasonic features of intra-amniotic "unidentified debris' at 14-16 weeks' gestation. AB - The objective of the study was to define the sonographic features of various echogenic materials which have been observed in the amniotic cavity of pregnancies in the early second trimester. The study population consisted of women who underwent a transvaginal ultrasound scan of pregnancy at 14-16 weeks' gestation. Our experience in 6500 examinations performed in both low- and high risk pregnancies is reported; 249 cases of intra-amniotic debris were identified in this population. Five different types of echogenic material were observed in the amniotic fluid: small floating particles, round structures, cystic structures, solid and semi-solid masses and highly echogenic material which looked like calcification. We conclude that intra-amniotic echogenic material may be observed during early pregnancy. Its origin and clinical significance need further evaluation. PMID- 8705410 TI - Doppler ultrasound of the uterine arteries: the importance of bilateral notching in the prediction of pre-eclampsia, placental abruption or delivery of a small for-gestational-age baby. AB - The use of Doppler studies of the uterine arteries in the prediction of pre eclampsia and intrauterine growth retardation has had mixed success. The introduction of color Doppler imaging and the use of the "notch' to define an abnormal waveform have helped to improve the predictive value of uterine artery Doppler screening. The aim of this study was to evaluate the use of uterine artery Doppler in a group of women of mixed race and parity. This study was a prospective, cross-sectional analysis of 1326 unselected women who were screened with continuous wave uterine Doppler at 19-21 weeks, as part of a fetal anomaly/dating scan. A total of 214 women with abnormal uterine artery waveforms (notching) were referred for assessment at 24 weeks; 191 attended and had color Doppler imaging/pulsed Doppler studies of both uterine arteries. Data from 185 pregnancies were suitable for analysis. There were abnormal uterine Doppler findings (uni- or bilateral notching) in 110 patients at 24 weeks; 48 had bilateral notching. The sensitivity of notching for the prediction of proteinuric pregnancy-induced hypertension (PPIH) was similar in primiparas (76.9%), multiparas (77.7%), African-Caribbean women (82.6%) and Caucasian women (71.4%). The sensitivity of bilateral notching for the prediction of PPIH requiring delivery before 34 weeks was 81.2%, and 57.6% for babies small for gestational age (SGA), with positive predictive values of 27% (PPIH), 31.2% (SGA) and 37.5% (any complication). Patients with persistent bilateral notching are particularly at risk of developing PPIH or delivering an SGA baby before 34 weeks' gestation; they warrant increased surveillance, and may be a group that could benefit from prophylactic therapies. PMID- 8705412 TI - Prenatal ultrasonographic fetal rib length measurement: correlation with gestational age. AB - The availability of normal data on fetal rib bone length would be of value in a variety of circumstances under which ultrasound examinations are performed. Fetal rib length measurements were obtained in 257 singleton fetuses in a prospective cross-sectional study. The study population consisted of pregnancies between 14 and 40 weeks with no evidence of growth disturbances, or structural or karyotypic abnormalities. Fetal rib length, as a function of gestational age, was expressed by the regression equation: RL = -0.5834 + 0.2030 (GA), where RL is the rib length in centimeters and GA the gestational age in weeks. The correlation was R = 0.94 (p < 0.0001). The normal limits of fetal rib length are defined, and a high correlation between fetal rib length, gestational age, and other standard determinants of fetal growth is demonstrated. Our results indicate that fetal rib length can be a useful adjunct in determining normal fetal growth and in the management of the pregnancy with fetal skeletal dysplasia. PMID- 8705411 TI - Doppler ultrasound imaging: a new technique to detect lung hypoplasia before birth? AB - Combined color-coded Doppler and pulsed Doppler ultrasonography allows visualization of the fetal pulmonary circulation and study of pulmonary blood flow velocity waveforms. Systolic and diastolic changes were observed in fetal pulmonary artery flow velocity waveforms in a case of fetal pulmonary hypoplasia at 34 weeks of gestation. This observation supports our hypothesis that Doppler velocimetry can detect lung hypoplasia based on post- mortem examination of decreased total size of the pulmonary vascular bed, decreased number of pulmonary vessels per unit lung tissue and increased pulmonary vascular muscularization. PMID- 8705413 TI - Transvaginal color Doppler studies in gestational trophoblastic disease. AB - Three women with gestational trophoblastic disease were examined using transvaginal color Doppler at the initial diagnosis and after each course of chemotherapy. In all cases, the examination before chemotherapy revealed hypoechoic areas surrounded by irregular echogenic areas and numerous intramyometrial flow signals. Pulsed Doppler examination revealed a low pulsatility index (PI) in the uterine arteries. After the completion of chemotherapy, a decrease in the vascularity and increase in the PI of the uterine arteries were demonstrated in two cases. These findings were closely consistent with the decrease in serum levels of beta-human chorionic gonadotropin. However, in the patient who developed permanent arteriovenous communication, hypervascularity persisted and the PI of the uterine arteries remained low, even after clinical remission was achieved. The present study indicates that transvaginal color Doppler ultrasonography is useful for the evaluation of gestational trophoblastic disease, both at the time of diagnosis and after chemotherapy. PMID- 8705414 TI - A comparison of patient tolerance of hysterosalpingo-contrast sonography (HyCoSy) with Echovist-200 and X-ray hysterosalpingography for outpatient investigation of infertile women. AB - The aim of this study was to assess patient tolerance of two outpatient tests. Sixty-six infertile women were prospectively randomized to hysterosalpingo contrast sonography (HyCoSy) (n = 34) or X-ray hysterosalpingography (HSG) (n = 32). The procedures were performed by the same operator. The uterine cavity outline and tubal patency were determined by both procedures. The mean times taken and the volume of contrast medium required for HyCoSy and HSG were similar: 12.1 +/- 5.2 and 9.5 +/- 4.8 min and 9.4 +/- 5.2 and 11.5 +/- 8.4 ml, respectively. Side-effects were assessed during the procedure, at 2 h, 24 h and 28 days. The most common side-effect was pelvic pain, in 56/66 (84%) women, occurring during the procedures (HyCoSy 19/34 (56%); HSG 23/32 (72/%)) and/or in the following 24 h (HyCoSy 14/34 (41%); HSG 15/32 (47%)). This was described as less severe or equal to their usual period pains (HyCoSy 100%; HSG 85%). Only 12/66 (18%) women required simple non-steroidal analgesia (HyCoSy 8/34 (24%); HSG 4/32 (13%)). There were no significant differences between the two methods concerning the frequency or severity of pains at different stages during and after the procedure or analgesia requirements. HyCoSy and HSG are equally well tolerated outpatient procedures for assessing tubal patency and uterine abnormalities. In addition, HyCoSy avoids the risks of ovarian irradiation and allows scanning of the uterine corpus and ovaries at the same time. PMID- 8705415 TI - Prenatal diagnosis of unilateral tibial hemimelia. AB - We describe a case of unilateral tibial agenesis which was initially observed at 21 weeks' gestation. Unlike bilateral tibial hemimelia syndrome, a rare autosomal dominant condition, unilateral tibial agenesis, which accounts for about three quarters of all newborns with this condition, has not previously been described. This case could have been a sporadic abnormality but, in view of the association with other observations (distal bifurcation of the femur, club foot), could be considered as an autosomal recessive inherited condition with variable penetrance. Ultrasonographic features and the genetic prognosis are discussed. PMID- 8705417 TI - Successful pregnancy after bilateral internal iliac artery ligation monitored by color Doppler imaging. AB - Ligation of the internal iliac (hypogastric) arteries can be a life-saving procedure in cases of severe hemorrhage from pelvic viscera. In young women it is the only way to preserve fertility when conservative methods have failed to control the bleeding. We present the case of a 21-year-old woman whose first pregnancy was terminated, after which profuse vaginal bleeding occurred and could not be controlled by conservative methods. An urgent laparotomy was performed and both internal iliac arteries were ligated. Two years after surgery, the patient conceived and after an uneventful pregnancy delivered a healthy infant. Uterine and fetal circulations were monitored with color Doppler imaging and were normal throughout pregnancy. A normal 3550-g male infant was delivered vaginally at 39 weeks' gestation. PMID- 8705416 TI - Prenatal ultrasound diagnosis of Roberts syndrome in a family with negative history. AB - A case is presented in which the ultrasonographic detection of multiple congenital anomalies led to the diagnosis of Roberts syndrome in the fetus of a woman with a negative family history. The fetus had bilateral cleft lip and palate, bilateral amesomelia with ectrodactyly, a complex congenital heart disease and intrauterine growth retardation. These malformations are frequent in Roberts syndrome and, therefore, an amniocentesis was performed to detect the cytogenetic marker of this syndrome, premature centromere separation. This phenomenon could not be detected in metaphases from amniocytes, but it was present in peripheral lymphocytes cultured at birth. The clinical implications of these findings are discussed. Furthermore, to our knowledge, this represents the first case in which the suspicion of Roberts syndrome was raised by ultrasound in a family with a negative family history. PMID- 8705418 TI - Intracardiac fibrin adhesive for selective fetocide in twin pregnancy: report of three cases. AB - Intracardiac fibrin adhesive was used for selective fetocide in three diamniotic twin pregnancies at 18-24 weeks' gestation (trisomy 21: n = 1; ultrasound evidence of major malformation: n = 2). Two of the three pregnancies were complicated by an intra-amniotic infection leading to pregnancy loss but one patient (with a monochorionic twin gestation) delivered a normal baby prematurely in the 31st week of pregnancy. The technique, complications and pregnancy outcomes are discussed. PMID- 8705419 TI - First-trimester ultrasound screening for chromosomal defects. PMID- 8705420 TI - Transient fetal ascites associated with Down's syndrome. PMID- 8705421 TI - Prenatal diagnosis and prognosis of a vein of Galen aneurysm assessed by pulsed and color Doppler sonography. PMID- 8705422 TI - [Aberrant biliary duct (vasa aberantia)]. PMID- 8705423 TI - [Aberrant drainage of bile duct]. PMID- 8705424 TI - [Sclerosing cholangitis (primary, secondary)]. PMID- 8705425 TI - [Duplication of extra-hepatic biliary duct (duplication of common hepatic duct, duplication of cystic duct, duplication of common bile duct)]. PMID- 8705426 TI - [Anomalies and variations of extrahepatic biliary tract]. PMID- 8705427 TI - [Congenital bile duct dilatation, congenital biliary dilatation]. PMID- 8705428 TI - [Congenital bile duct stenosis and biliary atresia]. PMID- 8705429 TI - [Cholecystohepatic ducts]. PMID- 8705430 TI - [Low junction of the cystic duct]. PMID- 8705431 TI - [Accessory bile ducts]. PMID- 8705432 TI - [Extra-hepatic cholangiolithiasis-choledocholithiasis]. PMID- 8705433 TI - [Mirizzi's syndrome, pseudo-lesion of bile duct due to the disorder of the cystic duct]. PMID- 8705434 TI - [Inflammatory polyp of the extra-hepatic bile duct]. PMID- 8705435 TI - [Syndrome of extrahepatic cholestasis, extrahepatic obstructive jaundice]. PMID- 8705436 TI - [Post operative bile duct stricture]. PMID- 8705437 TI - [Extrahepatic cholestatic syndrome in childhood]. PMID- 8705438 TI - [Anomalous arrangement of the pancreaticobiliary ductal system]. PMID- 8705439 TI - [Annular stricture of the common hepatic duct]. PMID- 8705440 TI - [Eosinophilic cholangitis]. PMID- 8705441 TI - [Septum formation of the common bile duct]. PMID- 8705442 TI - [Benign elevated lesions of the common bile duct]. PMID- 8705443 TI - [Extraductal bile duct obstruction, extraductal bile duct stenosis]. PMID- 8705444 TI - [Bile duct injury]. PMID- 8705445 TI - [Biliary dyskinesia, sphincter of Oddi dysfunction]. PMID- 8705446 TI - [Hemobilia]. PMID- 8705447 TI - [Gangrenous cholecystitis]. PMID- 8705448 TI - [Xanthogranulomatous cholecystitis]. PMID- 8705449 TI - [Metaplastic cholecystitis]. PMID- 8705450 TI - [Chronic non-suppurative destructive cholangitis (CNSDC)]. PMID- 8705451 TI - [Emphysematous cholecystitis]. PMID- 8705452 TI - [Acute cholecystitis]. PMID- 8705453 TI - [Ischemic cholecystitis]. PMID- 8705454 TI - [Postoperative acute cholecystitis]. PMID- 8705455 TI - [Cholecystitis]. PMID- 8705456 TI - [Tuberculosis of the gallbladder]. PMID- 8705458 TI - [Pericholecystic abscess]. PMID- 8705457 TI - [Isolated periarteritis nodosa of the gallbladder]. PMID- 8705459 TI - [Chronic cholecystitis]. PMID- 8705460 TI - [Acalculous cholecystitis]. PMID- 8705461 TI - [Lymph follicular cholecystitis]. PMID- 8705462 TI - [Gallbladder carcinoma producing CEA, AFP, CA19-9, sialyl SSEA and CSF]. PMID- 8705463 TI - [Klatskin tumor]. PMID- 8705464 TI - [Malignant melanoma of the gallbladder]. PMID- 8705465 TI - [Malignant fibrous histiocytoma of the gallbladder]. PMID- 8705466 TI - [Malignant lymphoma of the gallbladder]. PMID- 8705467 TI - [Inflammatory polyp]. PMID- 8705468 TI - [Oat cell carcinoma of the gallbladder]. PMID- 8705469 TI - [Hyperplastic polyp, metaplastic polyp]. PMID- 8705470 TI - [Granular cell tumor]. PMID- 8705471 TI - [Carcinoid of the gallbladder]. PMID- 8705472 TI - [Neoplasms of the cystic duct]. PMID- 8705473 TI - [Tumor of the extrahepatic bile ducts--benign, malignant]. PMID- 8705474 TI - [Carcinosarcoma and so-called carcinosarcoma of the gallbladder]. PMID- 8705475 TI - [Pseudopolyp of the gallbladder]. PMID- 8705476 TI - [Gallbladder neck cancer]. PMID- 8705477 TI - [Cholesterol polyp]. PMID- 8705478 TI - [Gallbladder tumors (benign and malignant)]. PMID- 8705479 TI - [Small polypoid lesions of the gallbladder]. PMID- 8705480 TI - [Fibroxanthogranulomatous inflammation]. PMID- 8705481 TI - [Fibroepithelial polyp of the gallbladder]. PMID- 8705482 TI - [Fibrous polyp of the gallbladder]. PMID- 8705483 TI - [Adenocarcinoma of the gallbladder]. PMID- 8705484 TI - [Adenoacanthoma]. PMID- 8705485 TI - [Adenomyomatosis of the gallbladder]. PMID- 8705486 TI - [Gallbladder adenoma (pyloric gland type, intestinal type, proper epithelial type, papillary type, mucinous cystic type]. PMID- 8705487 TI - [Carcinoma in adenoma of the gallbladder]. PMID- 8705488 TI - [Adenosquamous cell carcinoma of the gallbladder]. PMID- 8705489 TI - [Pleomorphic giant cell adenocarcinoma (PGCA-Ca)]. PMID- 8705490 TI - [Granulation polyp of the gallbladder, inflammation of the Rokitansky-Aschoff sinus]. PMID- 8705491 TI - [Gallbladder papilloma]. PMID- 8705492 TI - [Reactive lymphoid hyperplasia (RLH) of the gallbladder]. PMID- 8705493 TI - [Pseudotumors of the gallbladder]. PMID- 8705494 TI - [Leiomyoma, lipoma, myxoma, and fibroma of the gallbladder]. PMID- 8705495 TI - [Leiomyosarcoma of the gallbladder]. PMID- 8705496 TI - [Squamous cell carcinoma of the gallbladder]. PMID- 8705497 TI - [Lipoma and liposarcoma of the extrahepatic bile ducts]. PMID- 8705498 TI - [Undifferentiated carcinoma]. PMID- 8705499 TI - [Clear cell carcinoma of the gallbladder]. PMID- 8705500 TI - [Polyp and polypoid lesions of the gallbladder]. PMID- 8705501 TI - [Mucin producing gallbladder cancer]. PMID- 8705502 TI - [Congenital agenesis of the gallbladder, hypoplastic gallbladder, gallbladder atresia]. PMID- 8705504 TI - [Anomalous positions of the gallbladder]. PMID- 8705503 TI - [Heterotopic tissues in the gallbladder (heterotopic gastric mucosa, heterotopic intestinal epithelium, heterotopic pancreatic tissue and ectopic liver in the gallbladder]. PMID- 8705505 TI - [Formal abnormalities of the gallbladder]. PMID- 8705506 TI - [Neuroma of the extrahepatic bile duct, amputation neuroma of the bile duct]. PMID- 8705507 TI - [Gallbladder anomalies--multiple gallbladders]. PMID- 8705508 TI - [Variations in the arterial supply of the gallbladder]. PMID- 8705509 TI - [Anatomical variations of the gallbladder]. PMID- 8705510 TI - [Thickening of the GB wall]. PMID- 8705511 TI - [Cholelithiasis following gastrectomy]. PMID- 8705512 TI - [Cholecystolithiasis with ascariasis]. PMID- 8705513 TI - [Gas-containing gallstones]. PMID- 8705514 TI - [Confluence stone]. PMID- 8705515 TI - [Fibroma and fibrosarcoma of the extra hepatic bile ducts]. PMID- 8705516 TI - [Cholesterol gallstone--symptoms, diagnosis and treatments]. PMID- 8705518 TI - [Calcified gallstone]. PMID- 8705517 TI - [Pigment stone (black stones, calcium bilirubinate stones)]. PMID- 8705519 TI - [Cholecystocholedocholithiasis]. PMID- 8705520 TI - [Pseudostone of the gallbladder]. PMID- 8705521 TI - [Rare gallstone (calcium carbonate calculus)]. PMID- 8705522 TI - [Silent stone]. PMID- 8705523 TI - [Cystic duct and gallbladder remnant]. PMID- 8705524 TI - [Morgagni's hernia of gallbladder]. PMID- 8705525 TI - [Adenocarcinoma of the extra bile duct]. PMID- 8705526 TI - [Herniation of the gallbladder through the foramen of Winslow]. PMID- 8705527 TI - [Cholecystogastric fistula, cholecystoenteric fistula]. PMID- 8705528 TI - [Ulcer formation of gallbladder]. PMID- 8705529 TI - [Cystic duct syndrome]. PMID- 8705530 TI - [Cholesterosis (cholesterolosis) of the gallbladder]. PMID- 8705531 TI - [Hydrops of the gallbladder]. PMID- 8705532 TI - [Gallbladder injury, injury of the gallbladder, traumatic rupture of the gallbladder]. PMID- 8705533 TI - [Retention cyst of the gallbladder]. PMID- 8705534 TI - [Postcholecystectomy syndrome]. PMID- 8705535 TI - [Heterotopic bone formation]. PMID- 8705536 TI - [Intragallbladder ascariasis]. PMID- 8705537 TI - [Torsion of the gallbladder]. PMID- 8705538 TI - [Adenoacanthoma, adenosquamous carcinoma of the extrahepatic bile duct]. PMID- 8705539 TI - [Empyema of the gallbladder (empyema vesicae felleae)]. PMID- 8705540 TI - [Porcelain gallbladder]. PMID- 8705541 TI - [Spontaneous (idiopathic) perforation of the gallbladder]. PMID- 8705542 TI - [Parasitic infections in the extrahepatic biliary tree]. PMID- 8705543 TI - [Limy bile]. PMID- 8705544 TI - [Biloma]. PMID- 8705545 TI - [Biliary peritonitis]. PMID- 8705546 TI - [Extrahepatic cholangitis]. PMID- 8705547 TI - [Adenomyoma of the extrahepatic bile duct]. PMID- 8705548 TI - [Biliary fistula (postoperative, spontaneous)]. PMID- 8705549 TI - [Gallstone ileus]. PMID- 8705550 TI - [Biliary pancreatitis]. PMID- 8705551 TI - [Biliary sludge]. PMID- 8705552 TI - [Pneumobilia]. PMID- 8705553 TI - [Lipoma of the gallbladder]. PMID- 8705554 TI - [Cholecysto-choledocholithiasis]. PMID- 8705555 TI - [Carcinoma of the biliary tract]. PMID- 8705556 TI - [Inspissated bile syndrome]. PMID- 8705557 TI - [Parapapillary choledochoduodenal fistula]. PMID- 8705558 TI - [Adenoma, cyst adenoma of the extrahepatic bile duct]. PMID- 8705559 TI - [Extrahepatic intrabile duct tumor growth of hepatocellular carcinoma]. PMID- 8705560 TI - [Squamous cell carcinoma of the extrahepatic bile duct]. PMID- 8705561 TI - [Smooth muscle tumors of the extra-hepatic bile duct]. PMID- 8705562 TI - [Cancer of the main hepatic duct junction]. PMID- 8705563 TI - [Nodular-type carcinoma of extrahepatic bile ducts]. PMID- 8705564 TI - [Diffuse bile duct carcinoma]. PMID- 8705565 TI - [Invasive extrahepatic bile duct cancer]. PMID- 8705566 TI - [Acute obstructive suppurative cholangitis]. PMID- 8705567 TI - [Biliary papillomatosis]. PMID- 8705568 TI - [Special types of extrahepatic bile duct cancers]. PMID- 8705569 TI - [Papillary extrahepatic bile duct carcinoma, invasive papillary extrahepatic bile duct carcinoma]. PMID- 8705570 TI - [Mucin producing extrahepatic bile duct carcinoma]. PMID- 8705571 TI - [Intramucosal carcinoma of extrahepatic choledochus]. PMID- 8705572 TI - [Ectopic hormone producing cholangiocarcinoma]. PMID- 8705573 TI - Managed care and its relationship to oral and maxillofacial pathology. PMID- 8705574 TI - Ridge reconstruction with hydroxyapatite. PMID- 8705575 TI - Smokeless tobacco. PMID- 8705576 TI - Smokeless tobacco. PMID- 8705577 TI - INR and PTR anticoagulation values. PMID- 8705578 TI - Calcium channel blockers-induced gingival hyperplasia. PMID- 8705579 TI - Cause of median rhomboid glossitis. PMID- 8705580 TI - Bilateral static bone defects. PMID- 8705581 TI - Diagnosis of actinomycosis by fine-needle aspiration. PMID- 8705582 TI - Surgical management of ameloblastoma. Current status of the literature. PMID- 8705583 TI - A new therapeutic concept for the treatment of cystic hygroma. AB - A new therapeutic concept for the treatment of cystic lymphangioma (cystic hygroma) is presented. It consists of intralesional injection of triamcinolone (10 mg/kg) and surgical excision at a second stage if necessary. Four cases are presented to demonstrate the clinical application of this new therapeutic modality. Only in one case it was necessary to perform surgical excision as a second stage procedure, whereas three cases were successfully managed by intralesional injection of triamcinolone alone. PMID- 8705584 TI - A study of infrared thermographic assessment of liquid nitrogen cryotherapy. AB - OBJECTIVE: To assess whether infrared thermography can accurately predict the area of soft tissue necrosis that results from liquid nitrogen cryoprobe therapy. STUDY DESIGN: Eight rats received cryosurgery on the shaved abdomen with a liquid nitrogen cryoprobe in a triple-freeze technique. The therapy was monitored with infrared thermography and thermocouple probes. The temperature reached was correlated with the area of tissue necrosis found on histologic examination at sacrifice 1 week after the cryotreatment. A parallel study was carried out on pieces on beefsteak to assess the depth and shape of freeze. RESULTS: The -20 degrees C isotherm, which is felt to correspond to the cell lethal zone, occupied the inner 70% of the area of the iceball produced. Histologically, the -20 degrees C isotherm corresponded well to the area of tissue necrosis. In depth, the iceball takes on a semicircular shape. CONCLUSIONS: Infrared tomography is expensive to use clinically and cannot be readily used in the oral cavity. However, this study does show that one can clinically estimate that the inner 70% of the area of an iceball produced by liquid nitrogen on soft tissues will ultimately undergo necrosis. PMID- 8705585 TI - Expression of extracellular matrix in human mandibular condyle. AB - OBJECTIVE: The age-related expression of the extracellular matrices in human mandibular condyle was examined. STUDY DESIGN: The distribution patterns of types I to V collagens, laminin, fibronectin, fibronectin receptor, and transforming growth factor beta in 34 human mandibular condyles dissected from autopsy specimens were studied by immunohistochemical procedure with special attention on the age-related changes. RESULTS: Type I collagen was detected in the full layer of the condylar cartilage, and a stronger immunoreaction was delineated in the articular and cartilage zone. Types II and III collagen were mainly localized in the fibrocartilage zone. Type IV collagen and laminin were detected not only in the basement membrane of the blood vessels but also in the degenerated lesion where the expression of transforming growth factor beta was also detected. Immunostaining of type V collagen and fibronectin was noted in the perichondrocytic area, whereas that of fibronectin receptor was seen in the chondrocytes. In materials from younger cadavers types I, II, IV and V collagens, fibronectin, its receptor, and laminin showed stronger expression in the degenerative lesions than in the normal portions. In the sections from cadavers over the seventh decade, the immunoreaction of extracellular matrices was weak compared with the younger materials, and no increased reaction of extracellular matrices in the degenerative lesions was detected. In addition, severe osteoarthrosis was frequently seen in the older materials in macroscopic findings. CONCLUSION: These results suggest that the expression of extracellular matrices thus seems to be closely related to aging and degenerative changes in the condyle. PMID- 8705586 TI - Oral tuberculosis. AB - Tuberculous lesions of the oral cavity have become so infrequent that it is virtually a forgotten disease entity and may pose a diagnostic problem. Fifteen patients with conditions that were histologically diagnosed as oral tuberculosis were reviewed. All were men ranging in age from 29 to 78 years. The most common clinical presentation was odynophagia with a duration from less than 1 week to several years. The most frequently affected sites were the tongue base and gingiva. The oral lesions took the form of an irregular ulceration or a discrete granular mass. Mandibular bone destruction was evident in two patients. Two patients had a fever, and four had cervical lymphadenopathy. Eight cases were clinically suspicious for malignancy before biopsy. Only four patients had a history of tuberculosis, but 14 of the 15 patients were later found to have active pulmonary tuberculosis. Acid-fast bacilli were demonstrated in all patients. Tuberculosis should be considered in patients with an inflamed ulcer lesion. A biopsy specimen for histologic study, acid-fast stains, and cultures should be obtained for confirmation and differential diagnosis with other conditions. If a tuberculous lesion is suspected, a chest radiograph is indicated to investigate the possibility of pulmonary involvement. PMID- 8705587 TI - Helicobacter pylori in dental plaque and gastric mucosa. AB - OBJECTIVE: The aim of this study was to investigate the presence of Helicobacter pylori in both dental plaque and gastric mucus. STUDY DESIGN: Dental scaling hand instruments were used to collect supragingival and subgingival dental plaque from 81 dentate participants. Denture plaque was obtained from the fitting surfaces of dentures from 41 edentulous patients. Gastric mucus from gastric mucosa of antrum and body of stomach were collected from all 122 participants (92% white) with soft gastroscopic brush. These samples were dispersed in modified urea broth and normal saline solution before being inoculated onto selective Skirrow's agar and incubated in a microaerophilic atmosphere for culturing H. pylori. RESULTS: Dental plaque from all dentate participants was negative for H. pylori culture. Only one 80-year-old edentulous patient had positive H. pylori culture in both gastric mucus and denture plaque. CONCLUSIONS: Although dental plaque has a mixed flora that may act as a reservoir for gastric reinfection, dental plaque could not be implicated as the major reservoir of H. pylori for gastric reinfection. PMID- 8705588 TI - Trigeminal neuralgia versus atypical facial pain. A review of the literature and case report. AB - Trigeminal neuralgia and atypical facial pain are common conditions of facial pain. Although these two pain conditions are classically well separated in textbooks, a straightforward diagnosis may not always be possible because of the overlapping clinical signs and symptoms. In this article, a comparison and differentiation between the clinical and diagnostic features of these two pain conditions are presented. The general characteristics, etiologic characteristics, pathophysiology, differential diagnostic criteria, and therapeutic options of trigeminal neuralgia and atypical facial pain are described. A case report demonstrates the difficulties that can arise in the diagnosis and differentiation between the two disease entities. The article underscores the responsibility clinicians have in correctly diagnosing and managing patients with facial pain conditions. PMID- 8705589 TI - Herpesviridae-associated persistent mucocutaneous ulcers in acquired immunodeficiency syndrome. A clinicopathologic study. AB - Persistent mucocutaneous ulcers in AIDS represent a variety of disease entities. The purpose of this study was to characterize clinicopathologic features of persistent oral ulcers associated with cytomegalovirus and herpes simplex virus in AIDS. Forty-seven persons infected with HIV with persistent ulcers (mean, 2.4 ulcers/person) were included in this study. A biopsy specimen from a representative ulcer was taken from each patient. Hematoxylin-eosin, periodic acid-Schiff, cytomegalovirus, and herpes simplex virus immunocytochemical stains were performed on tissue sections. The most common sites of involvement were the buccal/labial mucosa (27%), tongue (25%), and gingiva (18%). Mean ulcer size was 1.8 cm with a mean duration of 5.6 weeks. The ulcerogenic viral agents were cytomegalovirus alone in 53% of cases, cytomegalovirus and herpes simplex virus coinfection in 28% of cases, and herpes simplex virus alone in 19% of cases. Treatment response to ganciclovir with or without topical steroids resulted in lesion resolution in the cytomegalovirus and cytomegalovirus/herpes simplex virus groups; however, recurrence/resistance was relatively high (23%). Herpes simplex virus/cytomegalovirus ulcers responded to oral acyclovir in combination with systemic ganciclovir. Increasing the oral acyclovir dosage resulted in resolution of herpes simplex virus-only ulcers in all but one case. Cytomegalovirus and herpes simplex virus are associated with persistent mucocutaneous ulcers in AIDS. These lesions responded to systemic antiviral therapy but are difficult to differentiate from other ulcerogenic diseases such as aphthous major, necrotizing stomatitis, and ulcerations not otherwise specified without biopsy and histopathologic examination. PMID- 8705591 TI - Malignant acanthosis nigricans with florid papillary oral lesions. AB - Acanthosis nigricans is a distinctive skin disease of importance, because it has served as an external marker for a variety of systemic disorders including endocrinopathies, and malignant tumors of internal organs. It typically appears as hyperpigmented, roughened plaques of velvety consistency and infrequently as verruca-like papillations. The oral cavity and lips can be affected by florid papillary growths. Because of its rarity and nonspecific microscopic appearance, clinical recognition of acanthosis nigricans continues to be a challenge. A case of mucocutaneous "malignant" acanthosis nigricans is presented in which pigmented skin lesions led to the discovery of a gastric adenocarcinoma, which in turn was followed by the appearance of massive oral papillomatosis. No effective treatment was found. PMID- 8705590 TI - Presence of human herpesvirus-like DNA sequence in oral Kaposi's sarcoma. A preliminary PCR study. AB - DNA sequences closely related to herpesvirus-like sequences have been found in AIDS-associated Kaposi's sarcoma. Subsequent studies indicate that these DNA sequences may be consistently found in AIDS Kaposi's sarcoma and non-AIDS Kaposi's sarcoma from different geographic regions. Oral Kaposi's sarcoma is rare and the specificity of this viral DNA in oral Kaposi's sarcoma and in other oral neoplasms has not been examined. A total of 77 oral neoplasms, including 3 Kaposi's sarcoma specimens from 2 patients with AIDS, were analyzed by polymerase chain reaction for the presence of this herpesvirus-like DNA sequence with the 330-233 primers. All three specimens of oral AIDS-Kaposi's sarcoma from two patients were positive for this DNA sequence. All of the other intraoral lesions including hemangioma, pyogenic granuloma, and various types of epithelial neoplasms, however, had negative reactions. Our results further confirm that oral Kaposi's sarcoma appears to be associated with this herpesvirus-like DNA sequence. PMID- 8705592 TI - Intraoral juvenile xanthogranuloma. A case report and literature review. AB - Juvenile xanthogranuloma rarely occurs in the oral cavity and has received little attention. A case of histologically documented juvenile xanthogranuloma of the oral cavity is described. This is the first intraoral case reported in the Oriental race and in the vestibule. Pertinent literature regarding intraoral lesions of this condition is also reviewed. PMID- 8705593 TI - True malignant mixed tumor (carcinosarcoma) of the parotid gland. Report of a case with immunohistochemical study. AB - We present a rare case of parotid gland carcinosarcoma that occurred in an 86 year-old woman. Clinical and histologic data revealed it had developed from a preexisting pleomorphic adenoma. Microscopic examination showed a neoplasm comprised of sarcomatous elements, mainly fibrosarcoma, intermingled with salivary duct-type carcinoma. Sarcomatous cells, excluding those that show chondrosarcomatous differentiation, reacted positively with antibodies directed against vimentin. Carcinomatous cells were positive for epithelial membrane antigen and keratin and negative for myoepithelial markers, which suggest an origin from salivary duct cells. This tumor seemed to consist of two histogenetically different populations of cells without evidence of a common origin from the myoepithelial cell. We also review the literature and discuss the histogenetic origin of true carcinosarcoma of salivary gland. PMID- 8705595 TI - Endodontic sequelae of miniplate bone fixation. AB - Various techniques of rigid internal fixation have evolved to achieve skeletal stability during healing. Although the miniplates and screws currently used have many advantages, their placement and presence may present significant difficulties for clinicians subsequently providing treatment. Problems with both endodontic diagnosis and treatment after miniplate fixation are illustrated with three case reports. PMID- 8705594 TI - Amalgam-associated oral lichenoid reactions. Clinical and histologic changes after removal of amalgam fillings. AB - OBJECTIVE AND STUDY DESIGN: Forty-nine consecutive patients with clinically diagnosed oral lichenoid reactions in contact with amalgam fillings were studied clinically and histologically. The long-term effect of replacement of these fillings was also examined. RESULTS: Seventeen (35%) patients showed positive reactions to mercury at the epicutaneous patch test that was carried out before treatment. After treatment, total regression of the lesions was found clinically in 33 (69%) and histologically in 26 (55%) patients. Most of the remaining lesions changed clinically and histologically to a less pronounced tissue reaction. Lesions in direct contact with amalgam fillings (group I) showed significantly better healing results than lesions that exceeded the contact area (group II). No difference in healing capacity was noted in the two groups between patients with positive patch reactions to mercury compared with those with negative reactions. Lesions that histologically were classified as benign oral keratosis showed a similar healing pattern as those classified as oral lichen planus. CONCLUSION: In group I all lesions changed histologically and clinically to a normal mucosa or to a less affected tissue reaction. In group II this change was less pronounced, which suggests that the fillings themselves were not the only factor involved in the cause of these lesions. The results suggest that various etiologic factors are involved in lichenoid reactions and that the effect of removal of amalgam fillings cannot be predicted by epicutaneous patch testing and biopsies. PMID- 8705596 TI - Dens evaginatus of anterior teeth. Literature review and radiographic survey of 15,000 teeth. AB - Case reports published between 1970 and 1995 that describe the characteristics of dens evaginatus of anterior teeth were reviewed. In addition, a radiographic survey of 15,000 anterior teeth was conducted to determine the occurrence of this developmental malformation. In this survey, dens evaginatus was found in about 1% of the anterior dentition and was more frequent in the maxilla, particularly in the lateral incisor. This anomaly was less frequent in the mandible. PMID- 8705597 TI - Radiopacity of potential root-end filling materials. AB - OBJECTIVES: In recent years various root-end filling materials have been suggested for clinical use. The purpose of this study was to assess the radiopacity of some potential materials according to ISO specification 6876. STUDY DESIGN: Radiographs were taken of 1-mm thick specimens of eight materials (amalgam, Kalzinol, IRM, Super EBA, Vitrebond, Fuji II LC, Chemfil, gutta percha); light transmission was assessed densitometrically and related to equivalent thickness of aluminum. RESULTS: Commercial glass ionomer cements (Vitrebond, Fuji II LC, Chemfil) had radiopacities below the international standard for root canal sealers (< 3-mm aluminum); three zinc oxide-eugenol cements (Kalzinol, Super EBA, IRM) had radiopacities equivalent to 5 to 8 mm aluminum; and gutta-percha had a radiopacity equivalent to 6.1-mm aluminum. CONCLUSIONS: We recommend that root-end filling materials should have a radiopacity greater than that for root canal sealers. PMID- 8705598 TI - Comparison of film, direct digital, and tuned-aperture computed tomography images to identify the location of crestal defects around endosseous titanium implants. AB - This study compared diagnostic performance obtained from two-dimensional and three-dimensional x-ray images. The latter were produced with a new tomosynthetic method based on aperture theory called tuned-aperture computed tomography. Seven human cadaver mandibular segments containing a total of 20 endosseous implants with a small randomly positioned alveolar crestal defect at each implant site were imaged in two dimensions with periapical film and with a charge-coupled digital detector, and digitally with the same detector in three dimensions with tuned-aperture computed tomography and subtracted tuned-aperture computed tomography techniques. Seven trained dentists viewed randomized displays of all modalities. Outcomes of the diagnostic task of identifying the locations of crestal defects were quantified with accuracy, confidence, and time performance measures. Analyses of variance demonstrated that differences between either three dimensional technique and either two-dimensional modality were significant for all measures (p < 0.001). These findings suggest that clinically applied TACT methods hold promise as an improvement over the status quo. PMID- 8705599 TI - Light source-induced error in computer-assisted image analysis with a video-based system. AB - OBJECTIVE: We tested the hypothesis that two different light sources, an alternating current fluorescent viewbox and a direct current halogen viewbox, do not differ with respect to their ability to illuminate reproducibly a radiograph during image capture. STUDY DESIGN: Two radiographs were taken: one with four hydroxyapatite chips mounted against a dry mandible and one without the chips. They were digitally subtracted with a video-based imaging system. The procedure was repeated at different times. RESULTS: A statistically significant difference among optical density measurements was found when the alternating current fluorescent viewbox (p < 0.001) was used and was related to light intensity variation. Such effect was not observed with the direct current halogen viewbox (p = 0.873). CONCLUSION: Study design efficiency was increased by 212% with the use of the direct current halogen viewbox so that to detect a specified treatment effect with a given level of statistical confidence, the sample size has to be 2.12 times greater if the alternating current fluorescent viewbox is used. PMID- 8705600 TI - Oral and maxillofacial radiology teaching file on the World Wide Web. AB - An oral and maxillofacial radiology teaching file on the World Wide Web was created effective June 15, 1995. The file contains 106 cases (279 images) of 94 diseases in the oral and maxillofacial region. PMID- 8705601 TI - Portrayals of nursing and the poetics of practice. PMID- 8705602 TI - Telling a story, writing a narrative: terminology in health care. PMID- 8705603 TI - Reclaiming 'Big Nurse': a feminist critique of Ken Kesey's portrayal of Nurse Ratched in One Flew Over the Cuckoo's Nest. AB - Nurse Ratched or 'Big Nurse' in Ken Kesey's counter-culture novel One Flew Over the Cuckoo's Nest is one of popular culture's most arresting and memorable images of the nurse. She is, however, deemed to be remarkable primarily for her malice and authoritarianism. This paper argues that such a purely realist reading fails to fully appreciate the significance of the character of Nurse Ratched. A feminist critique of the novel contends that the importance of 'Big Nurse' is less related to how realistic/unrealistic or good/bad she is as a nurse. Nurse Ratched is important because she exemplifies all that traditional masculinity abhors in women, and particularly in strong women in positions of power and influence. This paper explores the stereotype of 'Big Nurse' and argues that Kesey's vision of her ultimate 'conquest' is not a progressive allegory of 'individual freedom', but a reactionary misogyny which would deny women any function other than that of sexual trophy. PMID- 8705604 TI - Discipline and passion: meaning, masochism and mythology in popular medical romances. AB - This paper is an interpretive analysis of the discourses within popular romance literature, with a particular focus on the genre that includes constructions of the images of nurses and nursing. An historical contrast is made along with examinations of the uses and meanings encompassed within this body of literature, and its messages for women as nurses as it reflects/creates societal change. Deviations from the formulaic nature of these works are explored. Discipline and passion are the recurring themes evident throughout in juxtapositions of romance and power differentials in the hospital/medical scene. The soft-core, sado masochistic images evident in many titles are explored in a way that raises questions about sexuality, romance, nursing mythology, and the future of this sub genre of the popular romance. PMID- 8705605 TI - Narrative and exploration: toward a poetics of knowledge in nursing. AB - The dualism of subject and object has been a traditional model for nursing knowledge. That model is portrayed here as an epistemological exile. Our self imposed exile from the lived world of nursing can be remedied by inquiry based on engagement rather than distance. One model for engaged inquiry is explorers' journeys in remote regions. Knowledge of a region can be local or colonial, according to the explorer's stake in the region as homeland or territory. Nursing is an existential region where people live. In the exploration of nursing as homeland, knowledge takes the form of local narrative, the story of a place told by its inhabitants. Narrative inquiry offers nursing an epistemology that is both ethically and aesthetically congruent with its practice of engagement. PMID- 8705606 TI - The spirit of nursing and its healing art: a Jungian perspective. AB - Timely change is in the air as interest grows in the riches of nursing literature and other creative arts of nursing. Using Jungian and sociological concepts, this article takes a personal look at the literature's contribution to the profession. As the image of contemporary nursing is redefined, the power of the creative is evident, both in its grounding effect on individual practice and its unifying and strengthening potential for a changing profession. PMID- 8705607 TI - The art of stained glass: metaphor for the art of nursing. AB - The aesthetic is a way of knowing the meaning of and the meaning in the art of nursing. The art of creating stained glass offers a personal metaphor for nursing's essence; the art of caring. Both arts aim to fulfil the potential of their subjects to achieve a harmony that goes beyond their individual components. Stained glass artistry and caring in nursing require technical expertise, yet technical skill and knowledge are not the substance of either art. Both transcend space and time, and the art of stained glass and the art of nursing are influenced by the artist's/nurse's personal, social and cultural history. Just as the artisan transforms the glass and lead and is transformed in the creative moment, so does the caring transaction transform both patient and nurse. This personal reflection explores the nature of caring in nursing as mirrored by the author's work with stained glass. PMID- 8705608 TI - Our pictures of us. PMID- 8705609 TI - Giving permission to embodied knowing to inform nursing research methodology: the poetics of voice(s). AB - This paper originated from my experience of trying to find an authentic way to research women's experience of the premenstrum. I describe how personal change informed an evolving methodological approach. This change occurred when I felt tension between two strong voices. Conflict and insecurities originated from the pressure of my academic voice to conform to the dominant culture in what often seemed a disempowering way; a way that denied my body voice by emphasizing theory over experience. The construction of a poetic of voice(s) to illustrate how I eventually found an embodied voice is used to provide space for the reader's own interpretive process. The evolving embodied voice integrates my experience as a woman/nurse/researcher, validating my knowing bodily, practically, experientially and academically that I was able to help the participants to disclose their embodied knowing. This approach has wider implications when considering the value nurse researchers and academics give to embodied wisdom. PMID- 8705610 TI - Nurses, nursing and representation: an exploration of the effect of viewing positions on the textual portrayal of nursing. AB - Textual portrayals of nursing can be 'read' from a number of possible viewing positions, which play a very real part in the way that our reality is constructed and understood. The image depends on which viewing position is chosen by or constructed for the viewer, and how the viewer interacts with that position. Thus without the viewer the image is incomplete. This paper is a beginning exploration of the nature of texts that portray nursing, and what Kaplan has termed the 'delicate negotiation' between the reader and the text in producing, in this case, understandings of nursing. Consequently the discussion focuses on how texts represent nursing, not in terms of what they portray, but how understandings of that portrayal are arrived at. This is to move exploration of portrayals of nursing from the descriptive to the discursive realm. In so doing the aim is to begin to explore and uncover the way in which we are positioned, and position ourselves, as readers of portrayals of nursing in text. PMID- 8705612 TI - Transformational stories of healing in nursing. PMID- 8705611 TI - Signifying practices. PMID- 8705613 TI - (W)holism: philosophy and language (response) PMID- 8705614 TI - Nursing's agenda for change. PMID- 8705615 TI - The construction of a profession:a study of the history of nursing in Iceland. PMID- 8705617 TI - Rethinking cultural sensitivity. AB - The concept of cultural sensitivity is located within the tradition of anthropology and the history of colonisation and immigration in Australian society. This history provides a basis for examining the largely uncritical introduction of cultural considerations to the discipline of nursing. This paper argues that contemporary understandings of multiculturalism in nursing and health care policy tend to obscure, ignore and thus perpetuate notions of racial superiority. Recent works in transcultural nursing are used to illustrate the way in which ahistorical and therefore quite arbitrary traits are attributed to particular cultural groups. This perspective, given legitimacy in terms of cultural sensitivity, encourages political neutrality and thereby avoids questioning the discriminatory practices embedded in fundamental social relations. PMID- 8705616 TI - Containing abjection in nursing: the end of shift handover as a site of containment. AB - This theoretical paper has arisen out of a national study of the end of shift nursing handover. It draws upon and offers a postmodern reformulation of the psychodynamic concepts of anxiety and defence as articulated by Menzies Lyth in her study of nursing work in the 1950s. It outlines Kristeva's concept of abjection and draws upon Bion's formulation of the concept of containing to offer an account of how, in the current context of hospital nursing, the handover can function to contain abjection. PMID- 8705618 TI - Nurses, doctors and the body of the patient: medical dominance revisited. AB - Medical dominance is one of the most obvious features of the health care system and is particularly apparent in the relationship between doctors and nurses. Reasons given for the subordination of nurses to doctors have included matters of gender, class and state patronage of medicine. This paper, through an examination of university curricula and journal content, explores the notion that control over the model of the body is a major way in which medicine preserves its dominance over nursing. One reason for medicine maintaining its superior position is that nursing is not challenging this model of the body. PMID- 8705619 TI - The issue of critical paths in Australia: where are they taking us? AB - This discussion paper sets out some major concerns with the introduction of critical paths into the Australian health care system. It is argued that in their current form as devised in North America, critical paths may not be appropriate to our organisation of health care and may in fact be detrimental to the development of nursing practice. The discussion centres around the socio political context of the development of managed care in North America, as well as research findings from the implementation of critical paths in an Australian hospital. PMID- 8705620 TI - Nursing research and the philosophy of hermeneutics. AB - This paper explores some of the implications of hermeneutic philosophy for nursing research. Using illustrative material from recent research into the quality of life of older people in nursing homes, the paper shows that hermeneutics provides a perspective from which to examine the literature, and which guides both the collection and interpretation of data. The paper may be of interest to all scholars interested in the relationship between hermeneutics and the methods of social and nursing research. PMID- 8705621 TI - Norma Chick in a conversation with Judith Parker. Interview by Judith Parker. PMID- 8705622 TI - Family connections? Exploring nursing roles with families in home-based care. PMID- 8705623 TI - Why euthanasia? A reflective response. PMID- 8705624 TI - Are questionnaires so bad? PMID- 8705626 TI - Nursing's future. PMID- 8705625 TI - Managed care concepts for nurses. PMID- 8705627 TI - A vision for nursing education. PMID- 8705628 TI - Healthcare reform and the nurse executive. Interview by Marie Manthey. PMID- 8705629 TI - The role of theophylline in the treatment of asthma. PMID- 8705630 TI - Persistent sinusitis and refractory asthma in a 10-year-old boy. PMID- 8705631 TI - The "business" of medicine: hippocratic or hypocritical? PMID- 8705632 TI - Anti-inflammatory effects of theophylline: modulation of cytokine production. AB - BACKGROUND: The basis for the efficacy of theophylline in the treatment of asthma remains enigmatic. Although commonly classified as a bronchodilator, its ability to dilate smooth muscle is considered fairly poor and clinical responses are often independent of bronchodilation. Recent studies have suggested that immunomodulatory activities may contribute to the therapeutic benefit mediated by theophylline. OBJECTIVE: We performed these preliminary studies to determine whether theophylline modulates cytokine production by peripheral blood mononuclear cells. METHODS: Peripheral blood mononuclear cells were obtained from 24 asthmatic subjects and were left in a resting state or stimulated with either mitogens (phytohemagglutinin, lipopolysaccharide) or antigen (tetanus, cat) with or without the additional presence of theophylline (15 micrograms/dL). Supernatants were collected and evaluated for cytokine concentration by ELISA. RESULTS: Theophylline neither inhibited production of allergenic cytokines such as IL-4 nor modulated the repertoire of cytokines produced by TH cells. A statistically significant inhibition of spontaneous interferon-gamma synthesis was observed (24.5 +/- 8.6 to 13.4 +/- 4.2; P < .05). Theophylline did have anti inflammatory effects on cytokines primarily produced by mononuclear phagocytic cells. Theophylline mediated a slight inhibition of TNF-alpha production (0.26 +/ 0.08 to 0.21 +/- 0.06; P < .05). Theophylline was also associated with a 2.8 fold increase in spontaneous production of the anti-inflammatory cytokine IL-10 (0.35 +/- 0.08 to 0.98 +/- 0.16 ng; P < .01). CONCLUSIONS: A relative absence of IL-10 characterizes the asthmatic airways and may contribute to the development and severity of allergic inflammation. Induction of IL-10 production by theophylline may therefore mitigate inflammation and contribute to the clinical efficacy of this class of medications. PMID- 8705634 TI - The impact of managed care formularies on the practice of allergy-immunology. PMID- 8705633 TI - Latex hypersensitivity in emergency medical service providers. AB - BACKGROUND: Emergency medical service providers have a high degree of exposure to latex products. Patients utilizing emergency medical services can be allergic to latex products used during rescue efforts. OBJECTIVE: To determine the prevalence of latex hypersensitivity among emergency medical service providers. METHODS: Study questionnaires were distributed to a group of emergency medical service providers. Skin prick testing was performed using latex, common aeroallergens, and food extracts. Patch testing was done using latex and individual rubber additives. Serum latex-specific IgE was also measured. RESULTS: A total of 93 completed surveys were collected. Average exposure to latex in the work environment was 8.2 years. Eighty-four (90%) used latex gloves routinely at work. Of those, thirteen (14%) gave history of reaction to latex gloves. Sixty-two percent were not aware of the possibility of latex allergy in themselves or their patients. Forty-one (44%) had skin testing. Of those, four (10%) had positive prick tests for at least one of the four latex preparations used. Five had positive skin tests to avocado extract without supporting clinical history. Two had positive skin tests to banana, one with supporting clinical history for banana allergy. No food cross-reactivity with latex was demonstrated. Latex specific serum correlated with prick skin test results. No positive reactions were noted with patch testing. CONCLUSIONS: A significant percentage of emergency medical service providers were not aware of the occupational risk of latex allergy or the potential risk in their patients. A positive prick skin test for latex was present in 4 of 41 (10%), representing one-third of those who reported symptoms from latex exposure. PMID- 8705635 TI - Skin test results in related and unrelated persons with allergic rhinitis. AB - BACKGROUND: First degree relatives of patients with allergic rhinitis are at increased risk of having the disorder. It is not yet clear whether or not two such related persons with allergic rhinitis are more likely to be skin test positive to the same specific allergic extracts than unrelated persons with allergic rhinitis. OBJECTIVE: To determine whether or not first degree relatives (parent and child or full siblings) with allergic rhinitis are more likely to be skin test positive to the same allergen extracts than unrelated persons with the disease. METHODS: A retrospective chart review of Allergy Clinic patients was performed. Skin test results for 12 common aeroallergen extracts were compared in 32 pairs of genetically related subjects and 32 pairs of age-matched and sex matched unrelated subjects, all with allergic rhinitis. The percent of concordant results (both members of a pair skin test positive or both negative) and discordant results (one member of a pair positive and the other negative) were calculated and compared for related and unrelated groups. Also, the expected rates of positive concordance (both skin tests positive) based on the prevalence of skin test positively in the two groups were calculated and compared with the corresponding observed rates. RESULTS: Results were concordant in 242 of 384 skin tests (63%) in related pairs versus 233 of 384 skin tests (61%) in unrelated pairs (P = .28, not significant). The expected and observed rates of positive concordance were not significantly different in the related (P = .38) or unrelated (P = .49) groups. When the groups were subdivided into parent-child pairs and sibling pairs, a similar lack of significant differences between the groups was found. CONCLUSIONS: It is concluded that first degree relatives, both with allergic rhinitis, are no more likely to be skin test positive to the same allergen extracts than unrelated persons with this disease. Further, it seems appropriate to advise patients with allergic rhinitis that their first degree relatives who also have the disorder will not necessarily have the same specific allergies. PMID- 8705636 TI - Immunopathogenesis of fish allergy: identification of fish-allergic adults by skin test and radioallergosorbent test. AB - BACKGROUND: As the consumption of fish increases in the United States, the importance of allergic reactions to fish has become clear. Since most previous studies on fish allergy have focused on children reacting mainly to codfish, there is a need to investigate allergic reactions to other fish in adults. OBJECTIVE: To identify fish-allergic adults, and to assess cross-reactivity among different species of fish by RAST inhibition. METHODS: Thirty-nine individuals who reported fish allergy were selected for study; 32 (82%) were atopic as defined by two or more positive skin tests to common inhalant allergens and a history of allergic reactions and 33 (85%) experienced allergic symptoms within 30 minutes after ingesting fish. The most frequently reported symptoms were hives (69%), itching (69%), and wheezing/chest tightness (54%). Study subjects were skin tested with fish extracts and their sera assayed for IgE antibodies to different fish species. RESULTS: Thirty-six (92%) of the subjects tested had a positive skin test to at least one of 17 fish extracts tested; 9/35 (26%) reacted to all 17 extracts. Of the atopic (two or more positive skin tests to common inhalant allergens plus a personal and/or family history of allergy) and nonatopic fish-tolerant controls, 20/26 (77%) reacted by skin test to one or more fish extracts tested; the most prevalent positive reaction was to anchovy (73%). A significant correlation (P < .01) was observed between skin test reactivity of fish-allergic subjects to most fish extracts and fish RAST reactions. Radioallergosorbent inhibition testing demonstrated significant cross-reactivity among pollack, salmon, trout, and tuna; and between mackerel and anchovy. CONCLUSION: These results suggest that fish-allergic subjects may be clinically sensitive to more than one species of fish. Skin test reactivity to fish by itself is not an adequate criterion for the confirmation of clinically relevant fish allergy; consequently, fish-allergic subjects with positive skin tests to several fish species should exercise caution when eating fish until tolerance can be demonstrated by double-blind, placebo-controlled food challenge, at the patient's earliest convenience. PMID- 8705638 TI - Management of sinusitis in the asthmatic patient. AB - OBJECTIVE: To describe pathophysiologic links between sinusitis and asthma; to identify means of diagnosing sinusitis in the asthmatic; to discuss the management of asthmatic patients with sinusitis, both medical and surgical; to examine the outcome of managing the asthmatic patient with sinusitis. DATA SOURCES: Prospective and retrospective data from the author's experience was evaluated. Medline database was searched from January 1, 1984, using the keywords "asthma" and "sinusitis" without restriction to species or language; 48 articles identified. Relevant articles referenced in retrieved sources, current texts in otorhinolaryngology and sinus disease were also utilized. STUDY SELECTION: From data source abstracts, pertinent articles (33) and book chapters meeting the objectives of our paper were intensively reviewed. RESULTS: Clinical and experimental studies indicate that sinonasal inflammation can result in worsening of lower airway disease, while the exact nature of this relationship remains debated. Regardless of mechanism, identification of the asthmatic patient with chronic sinusitis using the techniques of nasal endoscopy and CT scanning can lead to treatment of sinusitis with overall sinus and asthmatic disease improvement. Proper management is first medical, while surgical approaches are reserved for persistent cases. Traditional, more radical, surgical approaches have shown good results overall, while newer techniques of functional endoscopic sinus surgery which respect anatomy and mucosal function are less studied but expected to result in similar or better long-term outcome. CONCLUSIONS: Sinusitis and asthma coexist and impact on one another at many different levels. Proper identification of the asthmatic patient with chronic sinusitis can be readily discerned by an accurate and thorough history and physical examination including nasal endoscopy and CT scanning. Proper medical and surgical management of sinusitis in the asthmatic patient can result in both improved sinonasal and asthmatic symptoms with fewer physician visits and decreased need for medication. PMID- 8705637 TI - Pathogenic role of thromboxane A2 in immediate food hypersensitivity reactions in children. AB - BACKGROUND: Food hypersensitivity, from the standpoint of pathogenesis as well as clinical management, remains controversial. During the food allergen-induced immediate hypersensitivity reaction, various chemical mediators are released. OBJECTIVE: The purpose of our study was to determine whether thromboxane A2 participates in food antigen-induced responses in children with food hypersensitivity. METHODS: Nine open food challenges were performed in nine patients with suspected food hypersensitivity. Plasma thromboxane B2 and histamine levels were measured during a 24-hour period following the challenge. RESULTS: All the patients demonstrated immediate reactions after food challenge. The mean plasma thromboxane B2 level (a marker of thromboxane A2 activity) rose significantly at two hours and three hours after the challenge. Simultaneously, the mean plasma histamine level rose significantly at two hours and three hours after the challenge. CONCLUSIONS: The results suggest that thromboxane A2 may play a pathogenic role in part in the immediate reaction after food challenge and that thromboxane A2 is probably released from a common cellular source (eg, mast cell) with histamine and/or by a common mechanism (eg, IgE-dependent platelet activation). PMID- 8705639 TI - Interferon-gamma and interleukin-4 production of ovalbumin-stimulated lymphocytes in egg-sensitive children. AB - BACKGROUND: The causal relation between egg allergy and cytokines derived from lymphocytes is unknown. OBJECTIVE: Interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) production of ovalbumin-stimulated and interleukin-2 (IL-2)-stimulated peripheral blood mononuclear cells from egg-sensitive patients was investigated and compared with that of stimulated peripheral blood mononuclear cells from nonatopic healthy children. METHODS: Peripheral blood mononuclear cells from egg sensitive patients and nonatopic healthy children were cultured with ovalbumin and IL-2 for seven days. The IFN-gamma and IL-4 concentrations in culture supernatants of the peripheral blood mononuclear cells were investigated. RESULTS: The levels of IFN-gamma production of only IL-2-stimulated or both ovalbumin-stimulated and IL-2-stimulated peripheral blood mononuclear cells from egg-sensitive patients with atopic dermatitis was significantly higher than that of healthy children and that of egg-sensitive patients with immediate allergic symptoms. CONCLUSIONS: Increased IFN-gamma production by lymphocytes after IL-2 and antigen stimulation has important implications for the mechanism of food sensitive atopic dermatitis. PMID- 8705640 TI - Isotype-specific antibody responses to acute Mycoplasma pneumoniae infection. AB - BACKGROUND: Mycoplasma pneumoniae-induced respiratory infections affect millions of patients and have been implicated in exacerbation of bronchial asthma. IgE may be involved in such exacerbations. While specific IgG and IgM responses to Mycoplasma pneumoniae are well described, the response of other isotypes is less known. PURPOSE: To determine whether specific IgE and what subclasses of IgG are formed in response to Mycoplasma pneumoniae infection. METHODS: We studied 20 patients with acute Mycoplasma pneumoniae infection, in whom the diagnosis was confirmed by a 16-fold increase in complement fixation titer between acute and convalescent serum samples. We developed an enzyme-linked immunosorbent assay (ELISA) for the determination of IgG, IgA, and IgM antibodies specific for Mycoplasma pneumoniae protein antigens. We used Western blotting to confirm the results of the ELISA and to detect Mycoplasma-specific IgG subclasses and IgE. RESULTS: Changes in Mycoplasma pneumoniae-specific IgG, IgA, and IgM were significant. Western blots of Mycoplasma pneumoniae antigens in 13 convalescent sera showed specific IgG in all, IgM in 11, IgA in 6, and IgE in 10. The IgG response consisted mainly of IgG1 and IgG3, and to a lesser degree of IgG2 and IgG4. CONCLUSIONS: We conclude that Mycoplasma pneumoniae infection is associated with a significant specific IgA and IgE response, in addition to the well-known responses of IgG and IgM. As IgE is involved in allergic reactions, the production of Mycoplasma pneumoniae-specific IgE may have a role in exacerbation of bronchial asthma. PMID- 8705641 TI - Efficacy of immunotherapy to ragweed antigen tested by controlled antigen exposure. AB - BACKGROUND: Immunotherapy is a recognized component in the management of allergic rhinitis. Its efficacy has been evaluated in a number of clinical field trials. These methods of evaluation are limited by control of antigen exposure. OBJECTIVE: A study was designed to evaluate the efficacy of immunotherapy in ragweed-induced rhinoconjunctivitis using an environmental exposure unit. METHODS: Forty-three subjects were grouped into (1) immunotherapy group: ragweed allergic subjects on maintenance ragweed immunotherapy for at least 2 years (N = 16), (2) positive control group: ragweed-allergic subjects who had never received immunotherapy (n = 16), and (3) negative control group: ragweed-nonallergic subjects (N = 11). Ragweed specific skin tests and ragweed IgE levels were obtained prior to exposure. The study was done in a room where levels of 2,500 to 3,000 grains m3 of ragweed were maintained over three hours. Symptoms were recorded every 15 minutes. RESULTS: Nasal symptoms in the immunotherapy group were significantly less than in the positive control group after 45 minutes (P = .025). Significant differences were not observed for ocular symptoms. Combined nasal and ocular scores were 50% less in the immunotherapy group than in the positive control group by 75 minutes (P = .039). Ragweed-specific skin tests and IgE were significantly less in the immunotherapy group than in the positive control group. Rhinoconjunctivitis symptoms in the negative control group were absent throughout. CONCLUSIONS: Controlled ragweed pollen exposure in this setting demonstrated that ragweed immunotherapy significantly reduced symptoms of ragweed-allergic rhinitis but had no significant effect on ocular symptoms. This system presents opportunities for additional studies on immunotherapy for allergic respiratory conditions. PMID- 8705642 TI - Asthma to carob bean flour. PMID- 8705643 TI - Asthma deaths. PMID- 8705644 TI - Allergy injection at home. PMID- 8705645 TI - Recognizing the management role of all nurses. PMID- 8705646 TI - HIV/AIDS: an imperative for a new paradigm for caring. PMID- 8705647 TI - Race, class, and gender considerations in nursing education. PMID- 8705648 TI - Is funding for graduate nurse education money wisely spent? PMID- 8705649 TI - The role of the international nurse in strategic planning. PMID- 8705650 TI - Esthetic and personal knowing through humanistic nursing. PMID- 8705652 TI - President's message: challenges and summons from within and without. PMID- 8705651 TI - De-gendered by pain. PMID- 8705653 TI - Conversations with CHAP: the role of benchmarks in home health care. PMID- 8705654 TI - Associate degree programs accredited by the NLN 1996-97. PMID- 8705655 TI - O come all ye faithful. PMID- 8705656 TI - Preparing acute care nurses for community-based care. PMID- 8705657 TI - Education for the nurse of tomorrow: a community-focused curriculum. PMID- 8705658 TI - The C word: a curriculum for the future. PMID- 8705659 TI - The changing health care environment: impact on curriculum and faculty. PMID- 8705660 TI - The NLN makes a new road. Interview by Catherine Bowles. PMID- 8705661 TI - Health care as performance art. PMID- 8705662 TI - President's message: from discipline specific to "inter" to "multi" to "transdisciplinary" health care education and practice. PMID- 8705664 TI - Elevated concentrations of interleukin-1 beta and interleukin-1 receptor antagonist in plasma of women with silicone breast implants. AB - Plasma from 27 women with silicone breast implants (SBIs) and 50 age-matched control women without SBIs were examined by enzyme immunoassay for the presence of interleukin-1 beta (IL-1 beta) and its naturally occurring receptor antagonist, IL-1ra. The results show that 74% (20 of 27) of women with SBIs had elevated concentrations of IL-1ra, whereas only 2% (1 of 50) of controls without SBIs had elevated concentrations of IL-1ra. In contrast to the IL-1ra results, the frequency of elevated IL-1 beta concentrations among women with SBIs was only 40% (11 of 27), but this was significantly higher than the 0% (0 of 50) in control women without SBIs. These findings suggest that there is a chronic ongoing inflammatory process in some women with SBIs, the implications of which are discussed in the context of silicone as an antigenic stimulant of the immune system. PMID- 8705663 TI - Immunodiagnosis of viral hepatitides A to E and non-A to -E. PMID- 8705665 TI - Effects of anticoagulant, serum, and temperature on the natural killer activity of human peripheral blood mononuclear cells stored overnight. AB - The effects of immediate versus delayed cell separation, storage temperature, presence of serum, and type of anticoagulation on the natural killer (NK) cytotoxicity of human mononuclear cells were assessed. The NK cytotoxicity of Ficoll-Hypaque-separated peripheral blood mononuclear cells (PBMC) was tested in a 3-h chromium-51 release assay with K562 cells at various effector/target cell ratios. The NK activities of PBMC from blood anticoagulated with either heparin or EDTA and then immediately separated and assayed were not different (42.9 +/- 2.5% for heparin and 40.3 +/- 4.6% for EDTA). When these separated cells were cultured in medium with 10% fetal calf serum and stored at 4,25, or 37 degrees C for 18 h before the assay, there was a significant increase in cytotoxicity. PBMC from blood stored in heparin or EDTA for 18 h before separation had reduced NK cytotoxicity, particularly if they were kept at 37 degrees C. When separated PBMC were cultured in medium with 10% human AB serum, however, samples held at 25 and 37 degrees C decreased in cytotoxicity but samples held at 4 degrees C maintained the cytotoxicity demonstrated at the baseline level with fresh cells. We recommend that heparinized blood be used for NK assays and that the PBMC be isolated immediately and held overnight at 4 degrees C in medium with 10% AB serum if the assay must be delayed. The NK cytotoxicity under these storage conditions most closely matches the results obtained when the PBMC are isolated and tested on the same day. IF PBMC isolation must also be postponed, it is best to store the blood in heparinized tubes at 25 degrees C to prevent loss of cytotoxic function. PMID- 8705666 TI - Peptide sequences of glycoprotein G-2 discriminate between herpes simplex virus type 2 (HSV-2) and HSV-1 antibodies. AB - The complete herpes simplex virus type 2 envelope glycoprotein G was represented by overlapping synthetic peptides. Herpes simplex virus type 2-specific human seroreactivities were mainly seen against three peptides, peptides G2-64, G2-69, and G2-70, located in the C-terminal part of glycoprotein G. This is, interestingly, a region which has strong homology between herpes simplex virus types 1 and 2. G2-69 was the most herpes simplex virus type 2-specific peptide, reacting with 93% (13 of 14) of herpes simplex virus type 2 immunoglobulin G positive human serum samples (n = 14) and none of the type 2 negative serum samples (n = 16) tested. The epitope of peptide G2-69 was mapped to the amino acid sequence RYAHPS, and the antibody binding to G2-64 could be increased by chemical oxidation of the peptide. The three reactive peptides were used, alone or combined, together with herpes simplex virus type 2 antigen in an attempt to discriminate anti-herpes simplex virus type 2 from anti-herpes simplex virus type 1 immunoglobulin G. Inclusion of the herpes simplex virus type 2 glycoprotein G peptides increased the specificity compared with the use of glycoprotein G alone. Combinations of peptides with whole glycoprotein also showed a greater discriminative capacity than single peptides. We conclude that these synthetic glycoprotein G peptides may be useful for herpes simplex virus type 2 serology based on peptides or combinations of peptides and antigens. This is the first study describing the possibility of discriminative herpes simplex virus serology by using synthetic peptides combined with small amounts of whole glycoprotein G. PMID- 8705668 TI - Evaluation of two Neospora caninum recombinant antigens for use in an enzyme linked immunosorbent assay for the diagnosis of bovine neosporosis. AB - Neospora caninum is a recently described apicomplexan parasite which causes paralysis and death in dogs. Neospora parasites also cause abortion and neonatal morbidity in cattle, sheep, goats, and horses, and neosporosis is emerging as an important cause of bovine abortion worldwide. The purpose of this study was to identify N. caninum cDNA clones encoding antigens that would be useful for the immunodiagnosis of bovine neosporosis. Two N. caninum tachyzoite cDNA clones expressing antigens that were recognized by serum from naturally and experimentally infected cattle were identified. The DNA sequences of these clones were determined, and the inserts were subcloned into the plasmid expression vector pTrcHisB. Both recombinant antigens, expressed as fusion proteins with a His6 tag, were purified on a nickel-chelating affinity column and evaluated in separate enzyme-linked immunosorbent assays (ELISAs). Both recombinant antigen ELISAs were capable of distinguishing between sera from Neospora-infected cows and sera from uninfected control cows. Furthermore, both assays were able to detect an antibody response in animals that were experimentally inoculated with N. caninum. Neither antigen showed evidence of cross-reactivity with serum from animals inoculated with the closely related parasites Toxoplasma gondii, Sarcocystis cruzi, Sarcocystis hominis, and Sarcocystis hirsuta. PMID- 8705667 TI - Cloning and expression of a putative alcohol dehydrogenase gene of Entamoeba histolytica and its application to immunological examination. AB - To clone and express the genes encoding major antigens of Entamoeba histolytica, we constructed a lambda gt11 cDNA library for E. histolytica HM1:IMSS and screened it with pooled sera from patients with amoebiasis. A 1,223-bp cDNA was cloned (clone 1223), and its nucleotide sequence was determined. The amino acid sequence predicted to be encoded by the open reading frame of clone 1223 consisted of 396 residues and showed 32.5 and 32.3% homology to the NADH dependent butanol dehydrogenases I and II (bdhA and bdhB) of Clostridium acetobutylicum, respectively. In addition, 29 of the 34 consensus positions of bdhA and bdhB were also well conserved in clone 1223. The recombinant protein expressed from clone 1223 had an estimated molecular mass of 43.5 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The antigenicity and specificity of the recombinant protein were evaluated by an enzyme-linked immunosorbent assay using sera obtained from two clinical groups of patients with amoebiasis and a group of healthy controls. The recombinant protein had potent and specific antigenicity. In all, 53 serum samples (88.3%) from 60 patients with amoebiasis were positive for immunoglobulin G antibody against the recombinant protein, with a mean optical density value of 0.42. In contrast, 53 of 54 healthy control serum samples were negative, with only 1 positive serum sample showing the lower optical density value. These results suggested that clone 1223 is promising in terms of providing a useful antigen for the accurate serodiagnosis of amoebiasis and that the gene encodes a putative alcohol dehydrogenase of E. histolytica. PMID- 8705669 TI - Neutralizing and enhancing activities of human respiratory syncytial virus specific antibodies. AB - The neutralizing and enhancing activities of respiratory syncytial virus (RSV) specific antibodies were examined. These two biological activities were measured for a panel of six monoclonal antibodies (MAbs) specific to the RSV surface F and G glycoproteins. Four MAbs specific for the F protein possessed both neutralizing and enhancing activities. One MAb (11-2-D2), specific to the G protein, enhanced RSV infection of U937 cells, a human macrophage cell line, but did not neutralize virus infectivity. One MAb (11-3-A3), specific to the F protein, efficiently neutralized virus infectivity but did not enhance RSV infection of U937 cells. MAb 11-3-A3 neutralized representative strains of the two antigenic subtypes of RSV. Assays performed with mixtures of MAbs showed that high concentrations of MAb 11-3-A3 masked the enhancing activity of MAb 11-2-D2. The assay of mixtures of two MAbs possessing only enhancing activities demonstrated that this response was synergistic. The role of neutralizing and enhancing antibodies in determining the outcome of RSV infection was examined for infants from whom cord blood serum samples were collected at birth. There was no significant difference in the magnitude of the serum-enhancing activities between infants who were hospitalized with RSV infections and a group of age- and sex-matched control infants with no reported respiratory illness requiring hospitalization. However, the results indicated a possible correlation between RSV infection of the infants and the occurrence of in vitro antibody-dependent enhancement of the cord blood sera at a serum dilution of 10(-2). A significant inverse correlation was found between the plaque-neutralizing and enhancing activities of the cord blood sera from infants, irrespective of subsequent RSV infection. These data are discussed in relation to the possible contribution of antibody-dependent enhancement to the normal course of RSV pathology in vivo. PMID- 8705670 TI - Utilization of tests for Lyme disease antibody at a university hospital. AB - We performed a retrospective study on patients who had a positive screening antibody test result for antibody to Borrelia burgdorferi to determine the clinical indicators used by physicians to order this test. Eighty-two evaluable patients who were screen positive (indirect enzyme-linked immunosorbent assay) between August 1991 and March 1993 were included. Additional tests, isotype specific capture immunoglobulin enzyme immunoassay and Western blot (immunoblot) analysis (immunoglobulin G), were performed on positive samples. Of 82 patients with a positive screening test result, 54 (66%) had no serologic evidence of Lyme disease on the basis of additional testing (positive predictive value, 34%). Only 28 of 82 patients (34%) had clinical indicators suggestive of Lyme disease. Antibody screening tests may provide misleading information if they are not accompanied by more specific assays. Inappropriate testing of patients without indications of Lyme disease is frequently performed, and the ordering practices of physicians should be reassessed. PMID- 8705671 TI - Detection of candidal antigens in autoimmune polyglandular syndrome type I. AB - Autoimmune polyglandular syndrome type I (APS I) is associated with chronic mucocutaneous candidiasis. To characterize the antibody responses in this subgroup of Candida albicans infections, we screened a candidal cDNA expression library with patient sera and found four cDNA clones encoding the immunopositive proteins enolase, heat shock protein 90, pyruvate kinase, and alcohol dehydrogenase. The reactivity to these antigens was studied further by immunoprecipitation assays with in vitro-transcribed and -translated proteins. Analysis of sera from 44 APS I patients showed that the highest antibody reactivity was found with enolase (80% of patients reactive), but significant serological responses were also found with heat shock protein 90 (67%), pyruvate kinase (62.5%), and alcohol dehydrogenase (64%). Overall, 95.5% of patients had detectable antibodies to at least one of these proteins. The cDNAs of enolase and heat shock protein 90 were also expressed in Escherichia coli and studied by immunoblotting. Again, 84% of sera reacted with enolase, whereas 44% of sera reacted with heat shock protein 90. A good correlation between the two methods was found for both enolase (r = 0.86; n = 58; P < 0.001) and heat shock protein 90 (r = 0.71; n = 56; P < 0.001). Our results indicate that the four abundant candidal proteins are the major antigens and can be used as accurate markers of candidiasis in APS I patients. The immunoprecipitation assay described here is particularly useful for the rapid analysis of a large number of samples. PMID- 8705672 TI - Determination of natural killer cell function by flow cytometry. AB - Natural killer cells (NK cells) are a subset of peripheral blood lymphocytes that mediate non-major histocompatibility complex-restricted cytotoxicity of foreign target cells. The "gold standard" assay for NK cell activity has been the chromium release assay. This method is not easily performed in the clinical laboratory because of difficulties with disposal of radioactive and hazardous materials, short reagent half-lives, expense, and difficulties with assay standardization. We describe a flow cytometric assay for the clinical measurement of NK cell activity. This study compared the chromium release assay and the flow cytometric assay by using clinically relevant specimens. There were no significant differences between the two assays in the measurement of lytic activity for 17 peripheral blood specimens or in reproducibility in repeated samplings of healthy individuals. We also established a normal range of values for NK activity in healthy adults and identified a small cluster of individuals who have exceptionally high or low levels of NK activity. The flow cytometric assay was validated by testing specimens from subjects expected to have abnormally low levels of NK activity (pregnant women) and specimens from healthy individuals in whom the activity of NK cells was enhanced by exposure to interleukin-2 or alpha interferon. Treatment with these agents was associated with a significant increase in NK activity. These results confirm and extend those of others, showing that the flow cytometric assay is a viable alternative to the chromium release assay for measuring NK cell activity. PMID- 8705673 TI - Utility of flow cytometric detection of CD69 expression as a rapid method for determining poly- and oligoclonal lymphocyte activation. AB - CD69 is a lymphoid activation antigen whose rapid expression (< or = 2 h postactivation) makes it amenable for the early detection of T-cell activation and for subset activation analyses. In the present study we evaluated the utility of flow cytometric detection of CD69 expression by T cells activated with polyclonal stimuli (anti-CD3 and staphylococcal enterotoxin B [SEB]) and oligoclonal stimuli (tetanus toxoid and allogeneic cells) using flow cytometry. Following activation of T cells with anti-CD3 or SEB, CD69 is detectable at < or = 4 h following activation, with anti-CD3 peaks at 18 to 48 h. Dose titration experiments indicated that CD69 expression largely paralleled that in [3H]thymidine incorporation assays, although the former offered a more sensitive measure of T-cell activation at limiting doses of activator than [3H]thymidine incorporation when cells were activated with either anti-CD3 or SEB. However, activation of T cells with either tetanus toxoid or allogeneic stimulator cells failed to induce detectable CD69 expression at up to 7 days of culture. Subset analyses of anti-CD3- and SEB-activated T cells indicated that populations other than T cells can express CD69 following stimulation with T-cell-specific stimuli, indicating that CD69 can be induced indirectly in non-T cells present in the population. These findings indicate that CD69 is a useful marker for quantifying T-cell and T-cell subset activation in mixed populations but that its utility might be restricted to potent stimuli that are characterized by their ability to activate large numbers of cells with rapid kinetics. PMID- 8705674 TI - Enzyme-linked immunosorbent assay with partially purified cytosoluble 28 kilodalton protein for serological differentiation between Brucella melitensis infected and B. melitensis Rev.1-vaccinated sheep. AB - The problem of differentiating sheep infected with Brucella melitensis from those vaccinated or exposed to cross-reaching organisms has not been resolved by conventional serological tests or through the use of the smooth lipopolysaccharide in primary binding assays. We therefore analyzed sera from ewes experimentally infected with B. melitensis H38, from ewes naturally infected with B. melitensis, and from B. melitensis Rev.1-vaccinated ewes by enzyme-linked immunosorbent assay with three antigenic fractions: O polysaccharide, a cytosoluble protein extract (CPE) from the rough strain B. melitensis B115, and a partially purified cytosoluble protein of 28 kDa (CP28) from the CPE. Immunoglobulin G anti-O polysaccharide and anti-CPE responses were detected in all groups of animals tested (Rev.1 vaccinated and B. melitensis infected). However, false-positive reactions with CPE occurred with sera from Brucella-free ewes. The use of partially purified CP28 abolished these false-positive reactions. Furthermore, no immunoglobulin G antibodies against CP28 were detected in sera from vaccinated ewes, whereas 80% (8 of 10) of ewes experimentally infected with B. melitensis H38 and 89% (25 of 28) of naturally infected ewes showed various degrees of anti-CP28 reactivity (absorbance values of between 0.5 and 2.5). The results obtained with CP28 showed the potential usefulness of this antigen to permit the detection of B. melitensis-infected ewes and their differentiation from B. melitensis Rev.1-vaccinated ones. PMID- 8705675 TI - Characterization of a monoclonal antibody specific for Brucella smooth lipopolysaccharide and development of a competitive enzyme-linked immunosorbent assay to improve the serological diagnosis of brucellosis. AB - The reactivity of monoclonal antibody (MAb) 12G12 was analyzed in regard to the main biovars of Brucella species and some members of the families Enterobacteriaceae and Vibrionaceae which present serological cross-reactions with the smooth lipopolysaccharide (S-LPS) of Brucella species. This MAb was strictly directed against the common specific epitope of the Brucella S-LPS. It recognized all of the smooth Brucella strains and biovars except B. suis biovar 2. In order to improve the specificity of the serological diagnosis of brucellosis, a competitive enzyme-linked immunosorbent assay (cELISA) was developed with the horseradish peroxidase-conjugated MAbs 12G12 and S-LPS of B. melitensis Rev1. The specificity of the cELISA was analyzed with 936 serum samples from healthy cattle. The assay was evaluated with sera from heifers (n = 18) experimentally infected with B. abortus 544. After infection, the performance of the cELISA was in agreement with those of the complement fixation test and the rose Bengal plate test. Finally, the specificity of the assay was also evaluated in regard to false-positive serological reactions by using sera from heifers experimentally infected with Yersinia enterocolitica 0:9 (n = 4) and with field sera presenting false-positive reactions (n = 74). The specificity of the cELISA was greater than the specificities of the complement fixation test and the rose Bengal plate test. Indeed, the new assay detected only 31 of the 101 false positive serum samples detected by at least one serological test. PMID- 8705676 TI - Novel enzyme-linked immunosorbent assays for the detection of anti-Fc gamma receptor autoantibodies. AB - There is a substantial interest in the role of antineutrophil antibodies since Fc gamma receptors (Fc gamma Rs) have been identified as the target for the majority of such autoantibodies. Antineutrophil antibodies have long been detected by an indirect immunofluorescence technique. Following optimization of the flow cytometric method of detection, we developed three enzyme-linked immunosorbent assays (ELISAs), each specific for autoantibodies against one of the three classes of human Fc gamma R. Fc gamma RI and Fc gamma RII were purified from cultured cells, and Fc gamma RIII was produced as a recombinant molecule. These were then used as capture agents in the respective ELISAs. When applied in parallel to a sizeable group of patients with primary Sjogren's syndrome, both methods established that anti-Fc gamma R autoantibodies were heterogeneous. This finding indicates that different populations of partly cross-reactive antibodies are detectable by these two methods. PMID- 8705677 TI - Immune responses to the R4 protein antigen of group B streptococci and its relationship to other streptococcal R4 proteins. AB - The R antigen, a trypsin-resistant protein observed in group A, C, F, G, and L streptococci, has also been found in group B streptococci (GBS). Although four species of the R antigen have been described for GBS, the R4 protein is the most prevalent in GBS isolates recovered from humans. This study examined the prevalence of antibodies against the R4 antigen by Western blot (immunoblot) (WB) in sera from 40 mothers colonized with GBS serotype II and III and from 26 noncolonized mothers; 92.5% of the colonized mothers had anti-R4 antibodies, compared with 54% of the noncolonized mothers (P < 0.001). Findings of antibodies in neonatal cord sera (n = 14) were concordant with maternal results by WB analysis for 71% of mother-infant pairs colonized with serotype II and for 57% of pairs colonized with serotype III. Of mothers known to be colonized with type II/R4 or III/R4, 100% (n = 12) had antibody against R4 by WB. This study also evaluated the prevalence of antibody to the GBS R4 antigen in 48 sera from individuals with high and low group A streptococcal anti-DNase B titers. Of those individuals with an anti-DNase B titer of > 640, 64% had a positive WB for anti R4 antibody, compared with 30% of individuals with low anti-DNase B titers (P < 0.05). The R4 antigen of GBS had immunologic identity to the R4 antigen of group A streptococci. Overall, the findings suggested that antibodies to the streptococcal R4 antigen were commonly present in GBS-colonized mothers and that transplacental passage of these antibodies occurred. The presence of antibody to R4 in non-GBS-colonized individuals may be due to immunologic responses to past exposure to the R antigen present in GBS or other streptococcal groups. PMID- 8705678 TI - Lymphocyte subpopulation reference ranges for monitoring human immunodeficiency virus-infected Chinese adults. AB - Two hundred eight healthy human immunodeficiency virus (HIV) type 1- and HIV type 2-seronegative Chinese adults (78 males and 130 females; mean age, 32 years; age range, 18 to 71 years) were analyzed for lymphocyte subsets by a standardized and quality-controlled flow cytometric immunophenotyping technique. While the leukocyte differential values were comparable to those found in studies of Caucasians, the means, medians, and 95% reference ranges of lymphocyte subsets were very different. The 95% reference ranges in absolute counts per microliter of whole blood (percentage of lymphocytes) for CD3+, CD3+ CD4+, CD3+ CD8+, CD3- CD19+ (B), and CD3- with CD16+ and/or CD56+ (NK) cells were 672 to 2,368 (54.8 to 83.0%), 292 to 1,366 (23.1 to 51.0%), 240 to 1,028 (17.9 to 47.5%), 82 to 560 (5.1 to 20.8%), and 130 to 938 (7.1 to 38.0%), respectively. CD3+ CD4+ cells showed significant sex difference (for males, mean of 702 [34.8%] and standard deviation of 258 [7.5%]; for females, mean of 728 [37.3%] and standard deviation of 254 [7.4%]) as well as an increase with age of 42 (1.6%) per decade. Investigations of the NK cell population did not show similar findings. Classification of HIV disease, treatment, and prophylactic regimens based on studies which relied heavily on estimations of lymphocyte subsets alone should be used with special caution for Chinese patients. Provided that adequate quality control measures are taken to ensure comparability of data, we recommend that these ranges be used on a day-to-day basis in laboratories that have not yet established their own reference ranges. PMID- 8705679 TI - Antigenicity of cell wall mannans of Candida albicans NIH B-792 (serotype B) strain cells cultured at high temperature in yeast extract-containing sabouraud liquid medium. AB - Cultivation of Candida albicans NIH B-792 (serotype B) at high temperature (37 degrees C) for 48 h in yeast extract-containing Sabouraud liquid medium (YSLM) provided the following findings in comparison with the findings obtained after incubation at 27 degrees C. Growth of the blastoconidia of this strain was decreased, with a dry weight of 9%, and the cells were deficient in cytokinesis. The cells did not undergo agglutination with serum factor 5 from a commercially available serum factor kit (Candida Check). Mannan (B-37-M) obtained from the cells cultured at 37 degrees C had partially lost its reactivity against serum factor 4 and lost most of its reactivity against serum factor 5 in an enzyme linked immunosorbent assay (ELISA) in contrast to that (B-27-M) at 27 degrees C. Both cells and mannan prepared by cultivation first at 37 degrees C and then at 27 degrees C entirely recovered their reactivities with serum factors 4 and 5. 1H nuclear magnetic resonance analysis also revealed that B-37-M had lost a beta-1,2 linked mannopyranose unit and retained a phosphate group. Similar changes were observed in the three other serotype B strains used in the study. The beta-1,2 linked mannooligosaccharides longer than mannotetraose were not included among the products released from B-37-M by mild acid treatment. The results of the inhibition ELISA with a series of beta-1,2-linked mannooligosaccharides from biose to octaose (M2 to M8, respectively) showed that the reactivity against serum factor 4 was inhibited most strongly by the oligosaccharides M4 to M8 and that the reactivity against serum factor 5 was inhibited completely by relatively longer oligosaccharides, M5 to M8, indicating their participation as the antigenic factor 5 epitopes. PMID- 8705680 TI - Detection of pulmonary and extrapulmonary tuberculosis patients with the 38 kilodalton antigen from Mycobacterium tuberculosis in a rapid membrane-based assay. AB - A rapid membrane-based serologic assay using the 38-kDa antigen from Mycobacterium tuberculosis for the diagnosis of tuberculosis (TB) was evaluated with 201 patients with pulmonary TB, 67 patients with extrapulmonary TB, 79 Mycobacterium bovis BCG-vaccinated healthy controls, and 77 non-TB respiratory patients. The overall sensitivities, specificities, and positive and negative predictive values were, respectively, 92, 92, 84, and 96% for sputum-positive TB patients; 70, 92, 87, and 79% for sputum-negative TB patients; and 76, 92, 80, and 90% for extrapulmonary-TB patients. Only 2% (1 of 44) of the healthy control BCG-vaccinated subjects gave weak positive signals in the assay, indicating that this rapid serological assay is a valuable aid in clinical diagnosis for both pulmonary and extrapulmonary TB. PMID- 8705682 TI - A monoclonal antibody reactive with a common epitope of Moraxella (Branhamella) catarrhalis lipopolysaccharides. AB - A hybrid cell line producing a monoclonal antibody (MAb) against Moraxella (Branhamella) catarrhalis lipopolysaccharide (LPS) was established. The specificity of the MAb 1B12 to purified rough LPSs from six strains of M. catarrhalis was ascertained by enzyme-linked immunosorbent assay (ELISA), competitive-inhibition ELISA, and immunoblotting. MAb 1B12 bound to live bacterial cells and culture supernatants from a total of 34 strains of M. catarrhalis, including 12 strains with different LPS serotypes. No cross reactions with smooth and rough LPSs from selected enterobacterial and nonenterobacterial strains, with other respiratory pathogens, or with Neisseria species were observed. These data suggest that MAb 1B12 recognizes a common epitope of M. catarrhalis LPS which differs from serotype determinants. PMID- 8705681 TI - Development of mucosal and systemic lymphoproliferative responses and protective immunity to human group A rotaviruses in a gnotobiotic pig model. AB - Gnotobiotic pigs were orally inoculated with virulent Wa strain (G1P1A[8]) human rotavirus (group 1), attenuated Wa rotavirus (group 2) or diluent (controls) and were challenged with virulent Wa rotavirus 21 days later. On various postinoculation or postchallenge days, virus-specific responses of systemic (blood and spleen) and intestinal (mesenteric lymph node and ileal lamina propria) mononuclear cells (MNC) were assessed by lymphoproliferative assays (LPA). After inoculation, 100% of group 1 pigs and 6% of group 2 pigs shed virus. Diarrhea occurred in 95, 12, and 13% of group 1, group 2, and control pigs, respectively. Only groups 1 and 2 developed virus-specific LPA responses prior to challenge. Group 1 developed significantly greater mean virus-specific LPA responses prior to challenge and showed no significant changes in tissue mean LPA responses postchallenge, and 100% were protected against virulent virus challenge. By comparison, both group 2 and controls had significantly lower LPA responses at challenge and both groups showed significant increases in mean LPA responses postchallenge. Eighty-one percent of group 2 and 100% of control pigs shed challenge virus, and both groups developed diarrhea that was similar in severity postchallenge. The virus-specific LPA responses of blood MNC mirrored those of intestinal MNC, albeit at a reduced level and only at early times postinoculation or postchallenge in all pigs. In a separate study evaluating antibody-secreting-cell responses of these pigs (L. Yuan, L.A. Ward, B.I. Rosen, T.L. To, and L.J. Saif, J. Virol. 70:3075-3083, 1996), we found that the magnitude of a tissue's LPA response positively correlated with the numbers of virus-specific antibody-secreting cells for that tissue, supporting the hypothesis that the LPA assesses T-helper-cell function. The magnitude of LPA responses in systemic and intestinal tissues also strongly correlated with the degree of protective immunity elicited by the inoculum (p = 0.81). We conclude that blood may provide a temporary "window" for monitoring intestinal T cells and that the LPA can be used to assess protective immunity to human rotaviruses. PMID- 8705683 TI - Recombinant Toxoplasma gondii surface antigen 1 (P30) expressed in Escherichia coli is recognized by human Toxoplasma-specific immunoglobulin M (IgM) and IgG antibodies. AB - The immunodominant surface antigen of Toxoplasma gondii, surface antigen 1 (SAG1), was expressed in Escherichia coli as a fusion protein containing a majority of the SAG1 protein supplied with six histidyl residues in the N terminal end. The recombinant protein was purified on a Ni-chelate column and then on a fast-performance liquid chromatography column and was in a nonreduced condition. It was recognized by T. gondii-specific human immunoglobulin G (IgG) and IgM antibodies as well as by a mouse monoclonal antibody (S13) recognizing only nonreduced native SAG1. Antibodies induced in mice by the recombinant SAG1 recognized native SAG1 from the T. gondii RH isolate in culture. Recombinant SAG1 is suitable for use in diagnostic systems for detecting anti-SAG1-specific IgG and IgM antibodies. PMID- 8705684 TI - The DNA-binding protein pUL57 of human cytomegalovirus: comparison of specific immunoglobulin M (IgM) reactivity with IgM reactivity to other major target antigens. AB - In this work we used PCR to amplify the DNA regions coding for two polypeptides from pUL57 of human cytomegalovirus (amino acids 540 to 601 and 1144 to 1233) and showed that both portions reacted very efficiently with immunoglobulin M in sera of acutely infected subjects. However, pUL57 is not an essential antigen for the replacement of or supplement to a cocktail of recombinant protein antigens containing portions of ppUL32, -44, -83, and -80a in immunoglobulin M serology. PMID- 8705685 TI - Time to peak serum antibody response to influenza vaccine in the elderly. AB - The earliest time at which serum antibody peaks following administration of influenza virus vaccine in elderly persons is not clearly defined. We compared the time intervals of 2 and 4 weeks after vaccination. A commercial trivalent vaccine containing the hemagglutinins of influenza viruses A/Texas/36/91(H1N1), A/Shangdong/9/93(H3N2), and B/Panama/45/90 was used. The hemagglutination inhibition antibody titers at 2 weeks after vaccination were identical to the hemagglutination inhibition antibody titers at 4 weeks for all three vaccine components. PMID- 8705687 TI - Determination of varicella-zoster virus (VZV) immune status with the VIDAS VZV immunoglobulin G automated immunoassay and the VZVScan latex agglutination assay. AB - The VIDAS varicella-zoster virus (VZV) immunoglobulin G immunoassay and the VZVScan latex agglutination (LA) test were compared for the detection of VZV antibody in patient sera. Of 625 samples tested, 554 were positive for VZV antibody by one or both methods. Five hundred thirteen (82.1%) samples were positive by both methods, 71 (11.4%) were negative by both methods, and there were 41 (6.6%) discrepant samples. Statistically significant differences in sensitivity and specificity were not found (P = 0.37); however, several observations were made. All 23 VIDAS repeat equivocal samples were LA positive. Fifty-four samples showed prozone effects with the LA assay, all of which resolved as positive. PMID- 8705686 TI - Antibody responses to defined epitopes in the Epstein-Barr virus BZLF1-encoded transactivator protein among human immunodeficiency virus-infected patients. AB - The Epstein-Barr virus BZLF1-encoded replication activator (ZEBRA) is a key mediator of reactivation from latency to the viral productive cycle. In the present study, the serum antibody responses against three defined ZEBRA epitopes (designated ZEBRA-1, -19, and -22) were determined for 50 human immunodeficiency virus (HIV)-seropositive patients and 100 matched healthy control subjects. The anti-ZEBRA responses were more commonly found among HIV-seropositive patients than among healthy controls for all the three ZEBRA epitopes tested (P < 0.0003, P < 0.003, and P < 0.001, respectively). Comparison of ZEBRA antibody levels with the degree of immunodeficiency (CD4 cell counts), CDC grouping, and HIV p24 antigen positivity showed little association, suggesting that induction of ZEBRA antibodies is an early event after HIV infection. PMID- 8705688 TI - [Immune function during space flights]. PMID- 8705689 TI - [Serum CA 19-9 levels in rheumatic diseases with interstitial pneumonia]. AB - Serum CA 19-9 (2-3 sialyl Le(a)) is a marker of malignant disorder such as pancreas or gall bladder cancers. It has been reported that sera from patients with interstitial pneumonia show elevated level of CA 19-9. To investigate the relationship between the elevation of serum CA 19-9 (sCA 19-9) and the presence of pulmonary fibrosis, we examined the level of sCA 19-9 in sera from patients with rheumatic diseases with or without interstitial pneumonia (IP). The sCA 19-9 level was determined by enzyme-linked immunosorbent assay (ELISA). Fourteen sera of 129 (10.9%) patients with rheumatic diseases without malignant disorders were positive for sCA 19-9 when normal range was determined as less than 100 U/ml (mean +/- 5 SD), and 26.7% of sera from poly/dermatomyositis (PM/DM) and 11.8% of systemic sclerosis (PSS) were positive for CA 19-9. Whereas only 8.0% of rheumatoid arthritis (RA) was positive. Twelve (28.6%) of 42 rheumatic patients with IP showed positive levels for sCA 19-9 (mean 142.5 +/- 363.0 U/ml), whereas only two (2.3%) of 87 without IP were positive (mean 33.9 +/- 65.8 U/ml; p < 0.05). The correlation between the level of sCA 19-9 and pulmonary diffusing capacity (%DLCO) revealed an inverse correlation in 32 rheumatic patients with IP (r = -0.43, p < 0.05). Furthermore, the elevated sCA 19-9 levels decreased after treatment with corticosteroid and/or cyclophosphamide or cyclosporin A. Therefore, elevation of the level of sCA 19-9 seems to be involved in the pathogenesis of IP and sCA 19-9 will be a useful parameter for IP. It has been reported that the CA 19-9 is produced from the bronchial glands and suggested that during chronic fibrotic process of the lung, the metaplastic change of the bronchial glandular cells occur and the cells produce CA 19-9. PMID- 8705690 TI - Interaction of indometacin farnesil, a new nonsteroidal antiinflammatory drug with peripheral blood mononuclear cells from patients with rheumatoid arthritis. AB - Indometacin farnesil (Indo-F) is a prodrug of indomethacin designed to reduce the occurrence of side-effects by esterification of the carboxyl group on indomethacin with farnesol. We have examined the pharmacological kinetics and action of Indo-F in peripheral blood mononuclear cells (PBMNC) and polymorphonuclear leukocytes (PBPNL) from patients with rheumatoid arthritis (RA). PBMNC and PBPNL were obtained from 31 RA patients. Indo-F was incubated with PBMNC or PBPNL in the presence or absence of granulocyte-macrophage colony stimulating factor (GM-CSF) (100 pg/ml) for 3 approximately 7 days, after which the concentrations of Indo-F and indomethacin in the culture supernatants or in the cytoplasm extracts were measured with HPLC. The levels of Indo-F in the culture supernatants were significantly decreased in the presence of PBMNC or PBPNL from either normal individuals or RA patients. Indo-F was found to be taken up by PBMNC as well as by PBPNL from RA patients. Conversion of Indo-F into indomethacin was significantly enhanced by GMCSF in the presence of PBMNC, but not PBPNL. The results indicate that Indo-F is taken up by peripheral blood leukocytes from RA patients. Moreover, the data suggest that monocyte-lineage cells might play an important role in the conversion of Indo-F into indomethacin since GM-CSF markedly facilitated the conversion in the presence of PBMNC, but not PBPNL. PMID- 8705691 TI - [Nasopharyngeal natural killer cell lymphoma with pericardial infiltration]. AB - A 77 year-old woman was admitted to the hospital because of nasal obstraction on March 1994. Tumorectomy of the nasopharyngeal tumor disclosed non-Hodgkin's lymphoma (LSG : diffuse, medium sized). The patient was treated with local radiotherapy to nasopharyngeal region and combined chemotherapy (2 courses of CHOP) to reduce residual tumor. On July, the pericardial effusion appeared and the large granular lymphocyte (LGL) like lymphoma cells were observed in the effusion. Flow cytometic analysis of these cells showed that they expressed CD 2, CD 7, CD 56 and HLA-DR, but did not express CD 3. T-cell receptor gene (TCR beta) rearrangement was not observed and natural killer activity was detected in these lymphoma cells. The patient was treated with ProMACE and the pericardial infusion of methotrexate, carboplatin and prednisolone, but the patient died of heart failure. Monoclonarity of lymphoma cells in the pericardial effusion was determined by southernblot analysis, using the terminal repeat of Epstein-Barr virus (EBV) for probe. It was suggested that EBV participated in tumorgenesis in this case. PMID- 8705692 TI - [Anti-platelet antibody and severe thrombocytopenia during interferon-alpha therapy for chronic active hepatitis C]. AB - We herein report a case of chronic hepatitis C where the patient developed severe thrombocytopenia during interferon therapy. The patient was a 61-year-old woman, who received interferon therapy on April 27, 1993 under the diagnosis of C type chronic active hepatitis. After 4 weeks, her platelet count had decreased to 18,000/microliters and intraoral hemorrhage had begun. Although she received 250 mg of methylprednisolone and 20 U of platelet transfusion three times, her platelet count continued to decrease to 4,000/microliters on both May 28, and on June 3, 1993, and so she was transferred to our hospital on June 4. On her second admission to our hospital, although the platelet-associated IgG (PA-IgG) had increased markedly and the megakaryocytes in her bone marrow had decreased, her platelet count had already increased to 37,000/ microliters, and this gradually returned to a normal level accompanied with a decrease of PA-IgG within one month In this case, although we found immunological abnormalities (high level of IgG, positive ANA and positive anti-smooth muscle antibody) prior to interferon treatment, we could not diagnose the patient as having suffered from autoimmune disease, including autoimmune hepatitis, because she did not satisfy the necessary criteria and because she did not have any symptoms suggesting autoimmune disease. We consider that there may be the possibility that interferon induced only an anti-platelet antibodies that caused the high level of PA-IgG and decreased the production level of platelets within the bone marrow. PMID- 8705693 TI - [Successful therapy with cyclosporine in a case with interstitial pneumonitis associated with polymyositis]. AB - We describe a case of interstitial pneumonitis associated with polymyositis who responded well to cyclosporine therapy. In October, 1993, a 49-year-old female was admitted to our hospital because of fever, muscle weakness and progressive dyspnea. Interstitial pneumonitis with polymyositis was diagnosed and daily oral steroid therapy was started. As the therapy was ineffective, pulse intravenous steroid therapies followed by oral cyclophosphamide and steroid therapy was given. Whereas serum levels of muscle enzymes decreased, she became progressively hypoxemic. No effect was observed in series of therapy including additional pulse steroid therapy, pulse intravenous cyclophosphamide therapy, oral azathioprine and oral bethametazone therapy. In April, 1994, oral cyclosporine was started. After daily oral cyclosporine was combined with steroid, her symptoms, chest X ray, chest CT and pulmonary function tests significantly improved. Her disease became stable and there was no signs of recurrence since then. It is suggested that cyclosporine is useful for the treatment of progressive interstitial pneumonitis associated with polymyositis. PMID- 8705694 TI - Reevaluation of enzyme-linked immunosorbent assay with ultraviolet-treated polystyrene microtiter plates for measurement of antibodies to dsDNA. AB - We reevaluated the efficacy of enzyme-linked immunosorbent assay (ELISA) with ultraviolet (UV)-treated polystyrene microtiter plates (UV-ELISA) for the detection of human serum anti-dsDNA antibodies. The subjects consisted of 38 patients with systemic lupus erythematosus (SLE). The titers of IgG serum anti dsDNA antibodies by UV-ELISA were significantly (p < 0.01) higher in sera from 26 active SLE patients than in those from 12 inactive SLE patients. ELISA with poly L-lysine coated microtiter plates (PLL-ELISA) revealed the similar results. However, the background ratio was only 1.0 +/- 0.9% by UV-ELISA, which was significantly (p < 0.01) lower than that by PLL-ELISA (16.8 +/- 10.8%). These results demonstrated the efficacy of ELISA with UV-treated polystyrene microtiter plates for the measurement of human serum anti-dsDNA antibodies in patients with SLE. PMID- 8705695 TI - Ethical issues in organ donation and transplantation: are we helping a few at the expense of many? AB - Ethical issues have always been apparent in the transplantation process and are becoming more evident as the demand for organs increases. The basic question is how just and ethical are the new policies enacted to encourage organ donation, considering that they affect the total public and benefit the small percentage of patients who require transplantation? Pros and cons of several of these policies will be discussed as will several clinical situations that raise ethical questions. PMID- 8705696 TI - Pastoral care in a critical care setting. AB - Healing the whole person-mind, body, spirit-cannot be forgotten in this world of critical care and high-technology medicine. It is the chaplain, one of many on the multidisciplinary care team, who is a reminder of this most important focus. PMID- 8705697 TI - Death notification: practical guidelines for health care professionals. AB - In acute care settings, death is common. Until now, few explicit guidelines for locating and notifying next of kin have been available. A licensed clinical social worker, in collaboration with two universities, surveyed medical professionals, university students in death and dying classes, and family members of newly admitted intensive care unit patients. From these responses, a working protocol for notifying and supporting the bereaved was developed. The general guidelines resulting from this study were applied and refined during the author's facilitation of more than 2,000 unanticipated death scenarios. Having the family view the body and talk with the medical staff is part of this protocol. PMID- 8705698 TI - Trauma bereavement program: review of development and implementation. AB - The Allegheny General Hospital Trauma Bereavement Program demonstrates the importance of specialized follow-up care for surviving family members and loved ones during the year after a death. Sudden, traumatic death leaves the survivors in shocked disbelief and intense emotional pain. Appropriate support and intervention can make a significant contribution to a family's eventual recovery by assisting in the normal grieving process and thus avoiding prolonged, pathologic grieving. The Allegheny General Hospital Trauma Bereavement Program assists a family as it begins to establish hope, order, and the promise of recovery. PMID- 8705699 TI - Assisted suicide: is there a future? Ethical and nursing considerations. AB - It is imperative that nurses take a personal and professional stand in the debate regarding physician-assisted suicide. Through case examples, this article defines and illustrates forms of euthanasia, including active and passive, voluntary and involuntary, double-effect, physician-assisted suicide, and physician aid-in dying. Ethical arguments against assisted suicide based on pain and suffering, quality of life, patient autonomy, and the common good are presented. The anticipated negative effects that legalized physician-assisted death would pose to the health professions and society are discussed. Guidelines and nursing implications conclude the article. PMID- 8705700 TI - Distributive justice and HIV disease in intensive care. AB - Human immunodeficiency virus (HIV) infection presents ethical dilemmas for health care professionals involved in critical care settings. Although critical care may prolong life for those with life-threatening illness or surgeries, the long-term outcome is still poor for those infected with HIV. Health care professionals have an obligation to provide quality care to prolong life. The requirement to prolong life in situations where the outcome is anticipated to be poor results in ethical conflict. The dilemmas created by the ethical principles that provide rational, systematic, critical, and principled thinking present multidimensional conflicts regarding how to live a moral life. The HIV epidemic has forced critical discussion about the allocation of health care dollars for the treatment of individuals who might not survive regardless of therapies. Allocation of care or distributive justice must be predicated on objective information that results in decisions that do not discriminate against a person with HIV infection. Ethically, sound decision making requires that all involved in the care and support of patients be a part of the process. PMID- 8705701 TI - Caring for the lesbian, gay, or bisexual patient: issues for critical care nurses. AB - Critical care nurses have a moral obligation to provide quality care for all of their patients. Many nurses lack information about lesbian, gay, and bisexual patient issues or feel uncomfortable working with such clients. This article explores some of the stereotypes that create negative attitudes about lesbian, gay, and bisexual people and offers suggestions for critical care nurses who wish to become better educated about these clients. PMID- 8705702 TI - How to lead effective teams. AB - Teamwork! Often teams are established to tackle tough jobs. Teams could focus on quality improvement, creating a new work process, or major problem solving of a current process. Effective leadership of the team is essential for reaching the team goal. How can team leaders be more effective? An excellent way to prepare for team leadership is to understand team management, leadership abilities, and characteristics of team effectiveness. This article discusses these important elements of effective teamwork. PMID- 8705703 TI - Trauma critical pathways: a care delivery system that works. AB - Continuous performance improvement in care a patient receives is of paramount importance to Robert Packer Hospital, a regional, accredited trauma center in rural Pennsylvania. In fall 1991, a care delivery system known as case management was introduced as a mode for achieving this continuous performance improvement goal. The staff members' experience with the initial development and use of a trauma critical pathway was first published in the November 1993 issue of Critical Care Nursing Quarterly. Now, 3 years later, case management has proven to be a viable means to accomplish the continuous performance improvement goal by enhancing the quality of care for trauma patients and their families throughout hospitalization. This article reviews the outcomes over the past 3 years as a result of case management and the use of trauma critical pathways. PMID- 8705704 TI - Kremer defends laser. PMID- 8705705 TI - Reporting of negative results important. PMID- 8705706 TI - The introduction of new procedures and technology: a different view. PMID- 8705707 TI - Closed-system technique for implantation of iris-supported negative-power intraocular lens. AB - BACKGROUND: The iris-supported claw lens can reversibly correct a wide range of ametropia and aniseikonia with excellent predictability and stability. However, concerns regarding its potential harm to the corneal endothelium have prevented its widespread use. Our experience with the closed-system surgical technique and the spreader device used in 180 cases between 1987 and 1993 has led us to conclude that potential complications are more likely due to the technique of implantation rather than to the properties of the lens itself. METHOD: We conducted a prospective study of 35 eyes of 20 consecutive patients (mean age, 38 years; range, 21 to 55 years) operated on between August 1993 and August 1994. Preoperative spherical equivalent refractions ranged from -6.00 to -21.25 diopters (D) (mean, -12.50 D). Follow up was 6 months (86% of eyes) to 1 year (57% of eyes). Spectacle-corrected visual acuity and endothelial cell density were measured prior to, and 1, 6, and 12 months following lens implantation. All data were analyzed using the paired t-test. RESULTS: Mean endothelial cell density dropped insignificantly (p > .10):by 1.22% at 1 month (n = 34), by 2.25% at 6 months (n = 27), and by 1.21% at 12 months (n = 18). Spectacle-corrected visual acuity increased significantly (p < .001): from a mean of 0.61 preoperatively to 0.77 at 1 month, 0.84 at 6 months, and 0.93 at 12 months postoperatively. At 12 months, 15 eyes (75%) had a refraction within +/-0.50 D, 19 eyes (95%) within +/-1.00 D, and 20 eyes (100%) within +/-2.00 D of emmetropia. CONCLUSIONS: Our results suggest that the closed-system approach using the spreader device allows safe implantation of the Worst iris-supported claw lens. PMID- 8705708 TI - Photorefractive keratectomy for myopia with a 6-mm beam diameter. AB - BACKGROUND: Earlier studies have reported clinical outcomes for up to 2 years following photorefractive keratectomy (PRK) using a 5.0-mm treatment zone with the Summit excimer laser. We present results of PRK using a 6-mm ablation zone with the same laser. METHODS: Forty-five eyes of 45 patients underwent excimer laser PRK for correction of myopia using a 6-mm excimer laser beam diameter. Attempted corrections ranged from 1.50 diopters (D) to 6.00 D. Data on outcomes of uncorrected visual acuity, spectacle-corrected visual acuity, predictability, corneal haze, subjective glare/halo effects, and patient satisfaction with the procedure were analyzed over a follow-up period of 6 months. RESULTS: All patients obtained a postoperative uncorrected visual acuity of at least 20/40; 28 eyes (62%) achieved at least 20/20. Postoperative spectacle-corrected visual acuity was at least 20/20 in all patients; no patients lost more than one Snellen line of spectacle-corrected visual acuity. Twenty-eight patients (62.2%) achieved within +/-0.50 D of the attempted correction; 40 patients (84.4%) achieved within +/-1.00 D. Mean spherical equivalent refraction was -4.99 D preoperatively and +0.44 D at 1 month, +0.04 D at 3 months, and +0.09 D at 6 months. At 6 months, 40 eyes (88.9%) were graded as clear, 4 eyes (8.9%) as having trace subepithelial haze, and 1 eye (2.2%) as having mild subepithelial haze. The mean glare/halo index for all of the patients was 0.59 on a scale of 0 to 5. Mean subjective patient satisfaction was 4.68 (on the same scale). CONCLUSIONS: Clinical outcomes following excimer laser PRK for myopia using a 6-mm treatment zone are encouraging. Postoperative subjective glare/halo were minimal, suggesting an optical advantage in using the larger ablation zone. PMID- 8705709 TI - Corneal asphericity and its implications for photorefractive keratectomy: a mathematical model. AB - BACKGROUND: Several clinical trials investigating myopic excimer laser photorefractive keratectomy (PRK) report an initial change in refraction from myopia to hyperopia, followed by a gradual regression toward emmetropia and occasionally to recurrent myopia. We examined the effect of corneal shape on refraction following PRK for myopia using a mathematical model. METHODS: We calculated the volume of corneal tissue removed by PRK for -3.00-diopter (D) and 6.00-D corrections with ablation diameters of 5 mm and 6 mm. For all the operating algorithms, the central region of the cornea was considered spherical. Mathematical models were developed based on calculations of the apical radius of the ablated cornea and the final refraction for a range of corneal asphericities. Baker's equation was used to model corneal asphericity. RESULTS: The smaller both the ablation size and desired correction, the smaller the effect of corneal asphericity on the refractive outcome. While corneal asphericity can influence the immediate refraction after PRK, the maximum effect is unlikely to be greater than +0.75 D. CONCLUSIONS: Corneal asphericity marginally affects the initial outcome of PRK. The effect will probably be offset by the healing response of the cornea. PMID- 8705710 TI - Comparison of corneal epithelial wound healing after photorefractive and lamellar keratectomy. AB - BACKGROUND: The rate of corneal epithelial wound healing may be determined, in part, by the characteristics of the stromal surface. The excimer laser has the ability to produce a highly uniform ablated surface, which may facilitate reepithelialization after photorefractive keratectomy (PRK). METHODS: The rate of corneal epithelial wound healing after excimer laser PRK was compared with the rate of reepithelialization after manual lamellar keratectomy. Ten rabbits received a 4-mm diameter ablation in one eye (fluence = 160 mJ/cm2) and a shallow, 5-mm diameter, manual lamellar keratectomy in the contralateral eye. At 0, 4, 8, 12, 24, 36, 48, 60, and 72 hours after wounding, sodium fluorescein was instilled, and photographs were taken, converted to video images, and digitized. Wound area was calculated for each time point and converted to wound radius; the slopes of the wound radius, plotted over time, were compared to determine rates of healing. Scanning electron microscopy was performed immediately after wounding to examine surface regularity. RESULTS: By 24 hours after wounding, corneas that had undergone PRK demonstrated a significantly faster rate of epithelial wound healing compared with eyes that underwent lamellar keratectomy (33.4 +/- 1.9 microns/hr vs 27.8 +/- 1.4 microns/hr, respectively, for 12 to 72 hours) (p < 0.0001). Scanning electron microscopy showed greater stromal surface irregularity in the corneas that had undergone lamellar keratectomy, compared with the laser ablated corneas. CONCLUSIONS: This study demonstrates that the rate of epithelial wound healing is significantly faster after excimer laser PRK than after lamellar keratectomy in the rabbit. Variations in surface regularity and wound edge profile may contribute to differences in wound healing. PMID- 8705711 TI - Control of pain after photorefractive keratectomy. AB - BACKGROUND: The inadequate control of pain after photorefractive keratectomy remains an important issue for both patients and clinicians. METHOD: A survey of postoperative pain regimens used in excimer centers worldwide was carried out. A pilot study to assess the temporal basis of pain after photorefractive keratectomy was done by using a visual analog pain score in 30 patients. A comprehensive literature search was undertaken to ascertain the suitability of topical ophthalmic anesthetics in controlling pain after photorefractive keratectomy. RESULTS: Many drugs, both oral and topical, are used worldwide to control pain after photorefractive keratectomy, but no group has shown satisfactory pain control in all patients. The pilot study revealed that the pain was most severe in the first 24 hours after photorefractive keratectomy and declined to low levels thereafter. Topical anesthetics cause corneal complications when used excessively and for prolonged periods. CONCLUSION: Pain after photorefractive keratectomy is most severe in the first 24 hours. Topical anesthetics would only be needed for this short time and would guarantee pain relief in all patients. We do not anticipate any corneal complications if the anesthetics are used for short times and under close medical supervision. PMID- 8705712 TI - Assessment of subjective pain following photorefractive keratectomy. Melbourne Excimer Laser Group. AB - BACKGROUND: Although researchers have reported that postoperative pain in patients undergoing excimer laser photorefractive keratectomy (PRK) surgery is common in the first 24 hours after excimer surgery, factors associated with increased postoperative pain have not been reported. The purpose of this study was to prospectively document self-reported pain following excimer laser surgery and explore associated factors. METHODS: Consecutive patients undergoing excimer laser photorefractive keratectomy (PRK) surgery to correct myopia and/or myopic astigmatism were asked to prospectively grade, on a four-point scale, the amount of pain they were experiencing immediately after treatment and again 2, 4, 8, 24, and 48 hours after surgery. They were also asked to record the type and dose of all medication taken during that time period. RESULTS: Pain questionnaires were returned by 62 patients (72%), ranging in age from 20 to 54 years. The mean self reported pain overall peaked 24 hours after treatment. Amount of myopia and prior excimer experience were not related to use of analgesia or self-reported pain (both p > 0.10). Patients who had three additional topical drops of indomethacin postoperatively reported significantly less pain 24 hours after treatment (t = 5.95, p = 0.0001). CONCLUSION: These results have implications for the education of patients about the likely course of healing after PRK. A course of five drops of topical indomethacin should be evaluated with a randomized clinical trial to assess efficacy in inhibiting ocular pain after PRK. PMID- 8705713 TI - Inflammatory cells, refractive regression, and haze after excimer laser PRK. AB - OBJECTIVE: To examine the role of inflammatory cell invasion and aspects of tissue reaction on refractive regression and corneal haze after myopic PRK in rabbits. METHODS: Measurements were made for 12 weeks postoperatively of haze intensity, corneal topography, tear cytology, inflammatory cell invasion, subepithelial fibroblast density, and the thickness of the newly laid down connective tissue and of the regrown epithelium. RESULTS: Inflammatory cell invasion could be prevented by fitting a soft contact lens, but this had no effect on the haze or the regression. Haze intensity correlated with subepithelial fibroblast proliferation but not with new connective tissue formation or epithelial hyperplasia. Neither connective tissue nor epithelium regrew in the form of a lens and thus they could not account for regression. Intensive treatment with corticosteroids resulted in a marked reduction in postoperative haze and a slight lessening of long-term regression. Correspondingly, it reduced fibroblast proliferation. CONCLUSIONS: Haze appears, at least in part, to be a result of fibroblast proliferation. Regression is possibly caused by slow distortion of existing tissue rather than the growth of new. PMID- 8705714 TI - Pilocarpine in the management of overcorrection after radial keratotomy. AB - BACKGROUND: Induced hyperopia is a potential complication of radial keratotomy with few effective treatments. We report the results of a retrospective study to evaluate the effectiveness of pilocarpine in the treatment of eyes overcorrected by radial keratotomy. METHODS: Sixteen eyes of 14 patients, from a consecutive pool of 200 eyes who underwent radial keratotomy, had hyperopia. The patients were subsequently treated with topical pilocarpine. The patients were treated from 3 to 14 weeks (mean, 8.2 weeks). RESULTS: The mean time of diagnosis of hyperopia was 3 weeks after the surgery (range, 1 to 12 weeks). The mean spherical equivalent of the manifest (fogging) refraction was +1.92 diopters (D) (range, +0.75 D to +5.00 D) and the keratometric power ranged from 31.25 D to 41.00 D (mean, 36.05 D). Mean uncorrected visual acuity before the treatment with pilocarpine was 20/50. After the treatment with pilocarpine, the mean spherical equivalent refraction was -0.31 D (range, -1.75 D to +0.50 D). The mean keratometric power was 38.32 D (range, 34.87 D to 43.12 D), with a mean uncorrected visual acuity at 20/25. The patients were followed for 8 to 49 weeks after treatment without pilocarpine (mean, 21 weeks). The mean spherical equivalent refraction and keratometric readings after that period were -0.71 D (range, -2.25 D to +0.25 D) and 38.33 D (range, 36.12 D to 43.12 D), respectively. All eyes in this study had more than 1.00 D of reduction of hyperopia at the conclusion of the study. CONCLUSION: Pilocarpine effectively reduced overcorrections after radial keratotomy. After termination of treatment, the steepening of corneal curvature was maintained. PMID- 8705715 TI - Acute tissue deformation of the human cornea after radial keratotomy. AB - PURPOSE: The regional deformation pattern of the cornea after radial keratotomy, which is essential for understanding the mode of action of the procedure, has not previously been studied in detail. METHODS: Up to 90 tiny mercury droplets were placed from center to limbus on the epithelial and endothelial corneal surfaces of eight eviscerated human donor eyes with four radial keratotomies (depth 100% of central corneal thickness, 3.5-mm clear zone). From digital images obtained under pressure loads ranging from 2 to 100 mm Hg, the distances between the fixed droplets were measured with an accuracy of 1 micron. After transforming the data to polar coordinates, regional meridional and circumferential strain patterns were calculated. Regional meridional and circumferential radii of curvatures were calculated from corneal profile images obtained at different pressure loads before and after keratotomy. RESULTS: Increasing the intraocular pressure from 2 to 100 mm Hg induced: an epithelial side wound gape of 44 mm; epithelial side circumferential tissue compression between incisions; considerable epithelial side meridional tissue elongation at and between incisions; little endothelial side circumferential strain across incisions; and little endothelial side meridional strain at and between incisions. The radial keratotomy induced 2.30 diopters (D) of central corneal flattening at an intraocular pressure of 2 mm Hg. The degree of central flattening correlated linearly with the amount of wound gape. In the physiological pressure range of the central cornea flattened 0.05 D for each millimeter-of-mercury increment in intraocular pressure. Pronounced meridional steepening was induced corresponding to the middle and peripheral parts of the keratotomy incisions. CONCLUSIONS: Our study suggests that the peripheral "tissue addition" seen after radial keratotomy is a net result of wound gape and circumferential tissue compression. Local bending of intact stromal tissue below the incisions plays an important role for the generation of the wound gape at the corneal surface. These data may help verify finite-element computer models of the human cornea. PMID- 8705716 TI - Excimer laser calibration system. AB - BACKGROUND: Excimer laser photoablation for refractive and therapeutic keratectomies has been demonstrated to be feasible and practicable. However, corneal laser ablations are not without problems, including the delivery and maintenance of a homogeneous beam. We have developed an excimer laser calibration system capable of characterizing a laser ablation profile. METHODS: Beam homogeneity is determined by the analysis of a polymethylmethacrylate (PMMA) based thin-film using video capture and image processing. The ablation profile is presented as a color-coded map. Interpolation of excimer calibration system analysis provides a three-dimensional representation of elevation profiles that correlates with two-dimensional scanning profilometry. Excimer calibration analysis was performed before treating a monkey undergoing phototherapeutic keratectomy and two human subjects undergoing myopic spherocylindrical photorefractive keratectomy. Excimer calibration analysis was performed before and after laser refurbishing. RESULTS: Laser ablation profiles in PMMA are resolved by the excimer calibration system to .006 microns/pulse. Correlations with ablative patterns in a monkey cornea were demonstrated with preoperative and postoperative keratometry using corneal topography, and two human subjects using video-keratography. Excimer calibration analysis predicted a central-steep-island ablative pattern with the VISX Twenty/Twenty laser, which was confirmed by corneal topography immediately postoperatively and at 1 week after reepithelialization in the monkey. Predicted central steep islands in the two human subjects were confirmed by video-keratography at 1 week and at 1 month. Subsequent technical refurbishing of the laser resulted in a beam with an overall increased ablation rate measured as microns/pulse with a donut ablation profile. A patient treated after repair of the laser electrodes demonstrated no central island. CONCLUSIONS: This excimer laser calibration system can precisely detect laser-beam ablation profiles. The calibration system correctly predicted central islands after excimer photoablation in a treated monkey cornea and in two treated human subjects. Detection of excimer-laser-beam ablation profiles may be useful for precise calibration of excimer lasers before human photorefractive and therapeutic surgery. PMID- 8705717 TI - Quantitative measurement of corneal haze after myopic PRK. AB - OBJECTIVE: To design and evaluate an instrument for the objective measurement of haze in rabbits after myopic PRK. METHODS: A circular fluorescent lamp bulb projected light onto the cornea through a circular collimating aperture covered with an orange filter. The image was collected on a centrally mounted CCD camera and the profile of the haze circle along its horizontal diameter determined. RESULTS: Haze circles were observed and quantitated. In the first weeks, they were donut-shaped, frequently with a central bright area, but later they became square-topped. The instrument reproduced measurements with an error of a few percentage points and showed good long-term stability. PMID- 8705718 TI - Photorefractive keratectomy and laser in situ keratomileusis for myopia between 6.00 and 10.00 diopters. AB - BACKGROUND: Excimer laser photorefractive keratectomy (PRK) can be effective in correcting myopia up to -6.00 diopters (D). Between -6.00 D and -10.00 D, the procedure is considered less effective and safe because it has been associated with dense scar formation and a high rate of regression. We compared photorefractive keratectomy (PRK) in this group of myopes with excimer laser keratomileusis in situ (LASIK). METHODS: Forty consecutive eyes with a manifest refraction between -6.00 and -10.00 D were treated with PRK using an ablation zone diameter of 6 mm. Subsequently, 40 consecutive eyes were treated with LASIK under a hinged flap using an ablation-zone diameter of 5 mm. All procedures used a Summit OmniMed laser and were done by the same surgeon. RESULTS: Preoperatively, 24 eyes (60%) undergoing PRK had 20/20 spectacle-corrected visual acuity; 1 year postoperatively, 20 (50%) had 20/20 vision uncorrected. Preoperatively, 13 eyes (33%) undergoing LASIK had 20/20 spectacle-corrected visual acuity; 1 year postoperatively, 24 (60%) could see 20/20 uncorrected. Sixteen (39%) PRK eyes had a spherical equivalent refraction within +/-1.00 D at 1 year; 20 (60%) eyes undergoing LASIK had a refraction within +/-1.00 D at that point. None of the eyes treated with LASIK developed corneal haze, while after PRK, 36 eyes (90%) developed haze (23 eyes [57%] +2 to +3). CONCLUSION: LASIK under a hinged flap proved superior to PRK in treating myopia between -6.00 D and -10.00 D. PMID- 8705719 TI - Learning from students. PMID- 8705720 TI - The reality of regional rates. PMID- 8705721 TI - Launching into election year. PMID- 8705722 TI - Making a difference to Maori health. PMID- 8705723 TI - Understanding wound healing. PMID- 8705724 TI - Putting principles into practice. PMID- 8705725 TI - Wound care. Case studies. PMID- 8705726 TI - Nursing behind bars. Interview by Anne Manchester. PMID- 8705727 TI - What price nursing education? PMID- 8705728 TI - Being called as a witness. PMID- 8705729 TI - A healthy proposal? PMID- 8705730 TI - Nursing older adults brings satisfaction. PMID- 8705731 TI - Reminiscences of Albert Sabin and his successful strategy for the development of the live oral poliovirus vaccine. PMID- 8705732 TI - Augmenting kidney mass at transplantation abrogates chronic renal allograft injury in rats. AB - Conventional renal transplantation, which substitutes a single allograft for two native kidneys, imposes an imbalance between nephron supply and the metabolic and excretory demands of the recipient. This discrepancy, which stimulates hyperfunction and hypertrophy of viable allograft nephrons, may be intensified by nephron loss through ischemia-reperfusion injury or acute rejection episodes occurring soon after transplantation. In other settings where less than 50% of the total renal mass remains, progressive glomerular injury develops through mechanisms associated with compensatory nephron hyperfiltration and hypertrophy. To determine whether responses to nephron loss contribute to chronic injury in renal allografts, nephron supply was restored to near-normal levels by transplanting Lewis recipients with two Fisher 344 kidneys (group 2A) compared with the standard single allograft F344 --> LEW rat model of late renal allograft failure (group 1A). At 20 weeks, indices of injury were observed in 1A but not 2A rats. These indices included proteinuria (1A: 45 +/- 13; 2A: 4.0 +/- 0.29 mg/day) and glomerulosclerosis (1A: 23 +/- 4.9%, 2A: 0.7 +/- 0.3%) (p < .05). Double allograft recipients maintained near normal renal structure and function, whereas 1A rats showed evidence of compensatory hyperfiltration (single-nephron glomerular filtration rate of 63 +/- 10 versus 44 +/- 2.0 nl/min in 2A rats) and hypertrophy (mean glomerular volume of 2.64 +/- 0.15 versus 1.52 +/- 0.05 microns3 x 10(6) in 2A rats) (p < .05). Thus, we conclude that a major component of late allograft injury is attributable to processes associated with inadequate transplanted renal mass, a finding that has major implications for kidney transplantation biology and policy. PMID- 8705733 TI - Specificity of circulating and tissue-bound autoantibodies in Goodpasture syndrome. AB - Goodpasture syndrome is an often fatal autoimmune disease associated with glomerulonephritis and/or pulmonary hemorrhage. The clinical manifestations of this disease correlate well with the presence of circulating antiglomerular basement membrane (GBM) autoantibodies. The primary target antigen in glomerular and alveolar basement membranes is thought to be the alpha 3 chain of type IV collagen. Nearly all that is known about anti-GBM antibodies in humans comes from work on unbound circulating antibody. We recently had the unique and rare opportunity to obtain early postmortem antibody and tissues from a patient who died with catastrophic Goodpasture syndrome. The specificity of circulating, kidney-bound and lung-bound autoantibodies from this patient was evaluated against a variety of purified basement membrane constituents. The results indicate that the primary target for the circulating and tissue-bound autoantibodies is the NC1 domain of the alpha 3(IV) chain of type IV collagen. Additionally, all the antibodies recognize a cryptic epitope/s on the alpha 3(IV)NC1 hexamer. Furthermore, tissue-bound and circulating antibodies compete with one another for overlapping epitopes on the antigen. These findings demonstrate that circulating autoantibodies in Goodpasture syndrome are highly representative of those bound to organ tissues, strengthening the notion that pathogenic autoantibodies are targeted to the alpha 3(IV)NC1 collagen, and that previous reports of findings in the circulation may be applicable to tissue injury. PMID- 8705734 TI - Stimulation and inhibition of human mammary epithelial cell duct morphogenesis in vitro. AB - We previously determined that normal human mammary epithelial cells (HMECs) placed on the basement membrane-like substance Matrigel form structures, whereas malignant breast cells do not (1). In the present study, we determined that the structures formed by normal cells on Matrigel resembled breast ducts in vivo by electron microscopy, and the process of their formation recapitulated what is known of duct formation in vivo. We therefore used this model to study less well understood aspects of breast morphogenesis. Two priming signals appeared necessary for initiation of morphogenesis: one provided by the Matrigel and one by the cells in an autocrine fashion. Evidence for this included diminished duct formation by cells plated low-concentration Matrigel or at low cell densities, and the reversal of the latter by conditioned medium from high-density cells on Matrigel. Antibodies to bFGF inhibited morphogenesis, suggesting a stimulatory autocrine role for this factor, and antibodies to TGF-beta 1 stimulated duct formation, suggesting an inhibitory autocrine role. Added TGF-beta 1 abolished morphogenesis and stimulated normal cells to wander through Matrigel as do malignant cells. Conditioned medium from normal cells did not stimulate malignant cells to form ducts, but conditioned medium from tumor cells diminished normal morphogenesis, suggesting that malignant cells secrete an inhibitor of morphogenesis. PMID- 8705736 TI - A model for keratinocyte gene therapy: preclinical and therapeutic considerations. AB - Gene transfer to the skin is essential for correcting genetic disorders and studying skin biology. Previous attempts at in vivo gene transfer have employed replication-deficient adenovirus injected subcutaneously and plasmid DNA propelled by a gene gun. In this report, we used mouse skin as a model and evaluated the efficiency of these two methods. Using the luciferase reporter gene, we found that adenovirus injected subcutaneously transduced primarily cells in the dermis. However, particle bombardment of skin by gene gun delivered the reporter gene mainly into the epidermis. When mouse skin was bombarded with a DNA construct expressing human TGF-alpha, the epidermis of the treated mice showed localized epidermal acanthosis and hypergranulosis, which resembled the histological phenotype of previously described transgenic mice overexpressing TGF alpha in the epidermis. These results suggest that the gene gun may be an effective tool for epidermal gene transfer and could be potentially useful in determining in vivo effects of growth factors and cytokines on the epidermis. PMID- 8705735 TI - Potential mechanism of estrogen-mediated decrease in bone formation: estrogen increases production of inhibitory insulin-like growth factor-binding protein-4. AB - Using a recently developed human osteoblastic cell line (hFOB/ER9) with high levels (approximately 4,000 per nucleus) of estrogen receptors and the characteristic phenotype of the mature osteoblast, we tested the hypothesis that estrogen decreases bone formation by inhibiting the action of the insulin-like growth factor (IGF) paracrine/autocrine system. IGF-II, the predominant IGF produced by osteoblastic cells, was measurable in hFOB/ER9-conditioned medium (approximately 10 ng/mL) and its level did not change significantly after treatment with 17 beta-estradiol (E2) or anti-estrogens. Treatment with E2 at 0.1 100 nM decreased [3H]thymidine uptake to 53% of control (p < 0.001) in a dose dependent fashion. The predominant IGF-binding proteins (IGFBPs) produced by hFOB/ER9 and by normal trabecular osteoblasts are IGFBP-3 and IGFBP-4, of which IGFBP-4 is consistently inhibitory of IGF action. Treatment with E2 at 0.01-10 nM for 48 h increased IGFBP-4 mRNA to 346% +/- 90% (mean +/- SE) of control (p < 0.05) and IGFBP-4 protein to 278% +/- 75% of control (p < 0.01) in a dose dependent fashion but did not alter IGFBP-3 mRNA or protein. E2 treatment also attenuated IGF-dependent, IGFBP-4 specific proteolysis to approximately 50% of control. ICI 182,780, a pure anti-estrogen, completely blocked E2-mediated decreases in cell proliferation and increases in levels of IGFBP-4 mRNA and protein. Treatment of the hFOB/ER9 cells with recombinant human IGFBP-4 (200 ng/mL) decreased cell proliferation to 55% of control (p < 0.01). Thus, E2 acts on osteoblastic cells to increase availability of inhibitory IGFBP-4, by both increasing its production and decreasing its degradation, which may oppose the mitogenic effect of the IGFs on osteoblastic cells. This action may mediate, at least in part, the decreases in bone formation that are observed after estrogen treatment in vivo. PMID- 8705737 TI - Effect of inhibitors of inducible form of nitric oxide synthase in infarcted heart muscle. AB - Nitric oxide (NO), an unstable radical, is synthesized from L-arginine by the constitutive (cNOS) and inducible (iNOS) forms of NOS. cNOS is present mainly in endothelial cells and plays a role in the regulation of blood flow. iNOS, the dominant enzyme in heart muscle during myocardial infarction, allograft rejection, and cardiomyopathy, is activated in macrophages. We recently described a significant increase of iNOS activity in macrophages of infarcted rabbit myocardium 24 hours after coronary occlusion, with peak activity occurring 3 days following coronary artery ligation. Inhibitors of NOS are L-arginine derivatives that inhibit both cNOS and iNOS; S-methylisothiourea (SMT) and aminoguanidine (AMG) are specific inhibitors of iNOS. Cyclosporin A and dexamethasone inhibit by interfering with protein synthesis. iNOS inhibition by SMT, NG-nitro-L-arginine (L-NNA), AMG, cyclosporin A and dexamethasone was examined in homogenates of normal, risk and infarcted myocardium. Three days after coronary artery ligation, the heart was excised and divided into normal, risk and infarcted regions. The inhibitory effect was calculated as IC50. Results shows that SMT was the most potent inhibitor with the lowest IC50; its effect, as well as the effects of L NNA and AMG, depended on the location in the myocardium. Inhibition for SMT and AMG was greater in the normal area than in the risk and infarcted regions. AMG induced an initial rise of iNOS followed by gradual decline in the area of risk and infarction. No inhibitory effects in cyclosporin A and dexamethasone were noted. PMID- 8705738 TI - Ultrasound biomicroscopic features of iris retraction syndrome. AB - BACKGROUND AND OBJECTIVES: To examine anatomic relationships of anterior and posterior chamber structures in iris retraction syndrome using ultrasound biomicroscopy. PATIENTS AND METHODS: Four eyes of four patients with iris retraction syndrome were imaged using high-frequency, high-resolution, anterior segment ultrasound biomicroscopy. In two patients, scans were obtained before and after medical therapy. RESULTS: Three patients presented with iris retraction configuration and one with iris bombe. In the latter case, iris bombe converted to iris retraction and back to iris bombe with the administration and later withdrawal of aqueous suppressants. Ciliochoroidal effusion was present in all eyes. CONCLUSIONS: Ciliochoroidal effusion is a constant feature of iris retraction syndrome. The hypotony associated with this disorder may result in part from aqueous hyposecretion related to ciliary body detachment. Ultrasound biomicroscopy is a useful aid in the diagnosis and follow-up of eyes with iris retraction syndrome. PMID- 8705739 TI - Corneal topographic changes and induced astigmatism resulting from superior and temporal scleral pocket incisions. AB - BACKGROUND AND OBJECTIVE: To determine the corneal topographic changes that are induced by superior and temporal 5-mm scleral pocket incisions. PATIENTS AND METHODS: The authors retrospectively reviewed the records of 43 patients who underwent phacoemulsification with posterior chamber lens implantation through unsutured 5-mm scleral pocket incisions, 24 superiorly and 19 temporally. Corneal curvature was measured using computerized videokeratography, and the induced astigmatism was calculated with the Holladay-Cravy-Koch formula. RESULTS: At 4 to 6 weeks, postoperatively there was mild flattening along with the meridian of the incision in both groups. The change was greater with the superior incisions, but the differences between the groups were not statistically significant. Surgically induced astigmatism was 0.7 D (+/-0.1 D) in the eyes that received superior incisions, and 0.3 D (+/-0.1 D) in the eyes that received temporal incisions; this difference was statistically significant (P < .03). CONCLUSIONS: At 4 to 6 weeks postoperatively, temporal 5.0-mm scleral pocket incisions provide more stable postoperative topography than do superior scleral pocket incisions. PMID- 8705740 TI - Use of cyanoacrylate tissue adhesive in small-incision cataract surgery. AB - BACKGROUND AND OBJECTIVE: A controlled clinical study was performed on three different groups of patients. PATIENTS AND METHODS: Each patient had a 6.5 mm wide scleral tunnel with phacoemulsification. Sealing of the incision was done either without suture, with an anchor suture, or with tissue adhesive (cyanoacrylate). The authors observed whatever induced astigmatism was present during the 1st and 12th weeks of the postoperative period. RESULTS: In the immediate postoperative period, astigmatism was significantly less (P < .01) in the groups of patients with suture and tissue adhesive than in the unsutured group. The results after 12 weeks were similar for the three groups (P > .05), with no complications observed as a result of the use of cyanoacrylate. CONCLUSION: Cyanoacrylate seems to be a promising innovation in scleral tunnel surgery for cataracts. It is an effective, a quick, and a safe alternative to sutures. PMID- 8705741 TI - Retrobulbar anesthesia with a flexible catheter. AB - BACKGROUND AND OBJECTIVES: The authors describe a method that seeks to improve the administration of local anesthesia for intraocular surgery, avoiding the risk of potential complications associated with retrobulbar and peribulbar techniques and eliminating the inconveniences of the new methods such as subconjunctival and limbal infiltration. PATIENTS AND METHODS: The authors demonstrate the safety and effectiveness of this technique in 50 consecutive cases. This technique consists of a small, blunt dissection of the conjunctiva, Tenon's capsule, and intermuscular septum in the inferior nasal quadrant, followed by the insertion of a flexible catheter that is guided to the retrobulbar space, where 3.0 ml of anesthetic solution is injected. RESULTS: All of the patients (100%) felt that this procedure was comfortable and painless. No complications were observed. Ultrasonography demonstrated the exact catheter position in the retrobulbar space; therefore, the authors could be sure that this anesthetic infiltration was done in the appropriate and safe place. CONCLUSIONS: This is a direct sub-Tenon's technique that is simple, effective, and safe and that avoids introducing a sharp or metallic instrument into the orbit. PMID- 8705742 TI - Fluorescent vesicle angiography with sodium fluorescein and indocyanine green. AB - BACKGROUND AND OBJECTIVE: The authors evaluated the feasibility of merging free dye angiography and the fluorescent vesicle technique to achieve the best characteristics of both. MATERIALS AND METHODS: Fluorescent vesicles encapsulated with either indocyanine green or carboxyfluorescein were mixed with free indocyanine green or free sodium fluorescein, respectively, and imaged with a scanning laser ophthalmoscope in both an in vitro model and primate and rabbit models. RESULTS: In the in vitro model of the sodium fluorescein combination, optimal viewing of vesicleen capsulated dye and free dye was at a ratio of 150:1; for indocyanine green, the ratio was 50:1. In vivo, high-quality fluorescent vesicle angiograms were obtained that demonstrated leakage of free dye from choroidal laser spots. CONCLUSIONS: Free dye and fluorescent vesicles can be combined to obtain an angiogram with all of the advantages of a traditional angiogram, while allowing the operator to assess the changes in retinal or choroidal circulation directly. PMID- 8705743 TI - Comparison of argon laser iridotomy and sequential argon laser and Nd:YAG laser iridotomy in dark irides. AB - BACKGROUND AND OBJECTIVE: A prospective study was performed to compare argon laser iridotomy and sequential argon laser and Nd:YAG laser iridotomy in dark irides. PATIENTS AND METHODS: Twenty-four eyes of 17 patients underwent laser iridotomies; 13 underwent argon laser iridotomies and 11 underwent sequential argon laser and Nd:YAG laser iridotomies. The argon laser settings were standardized at 1.2 W, 50-mum spot size, and 0.1-second duration. The Nd:YAG laser was set at 2.5 mJ and single-pulse shots were used. RESULTS: All of the iridotomies were patent at the end of one treatment session. The mean total energy used for argon laser iridotomy was 8.28 J. For sequential iridotomy, 3.12 J was used for the argon laser stage and 7.5 mJ for the Nd:YAG laser stage. CONCLUSION: The total argon laser energy used can be reduced by 2.65 times by using the sequential laser iridotomy technique. PMID- 8705744 TI - Argon laser peripheral iridoplasty. PMID- 8705745 TI - Experimental nonpenetrating transscleral cyclodiathermy in rabbits. AB - BACKGROUND AND OBJECTIVE: Acute and long-term effects of contact transscleral cyclodiathermy (CTCD) were studied in rabbits. MATERIALS AND METHODS: In the acute phase, three evenly spaced applications were placed at the limbus in each of four quadrants using the Ophthalmic Diathermy TR4000 (MIRA, Inc., Waltham, MA). Each quadrant of 17 eyes of 13 Dutchbelted rabbits was treated with radio frequencies (RFs) of 1.0 to 4.5, for times of 0.5, 0.75, 1.0 to 3.0, and 4.0 to 10.0 seconds. Eyes were then immediately enucleated and fixed in Karnovsky's solution after the rabbits had been sacrificed with 3 ml of pentobarbital sodium. In the longitudinal phase, four groups of six rabbits each were treated with 20 evenly spaced applications at the limbus at one of four settings: 1.5 RFs/4 seconds, 2.0 RFs/3 seconds, 2.5 RFs/2 seconds, or 3.0 RFs/2 seconds. Intraocular pressures were measured on alternate days for 1 week prior to treatment and for 4 weeks after treatment, following which the eyes were enucleated and fixed in Karnovsky's solution. RESULTS: Gross examination of the acute phase eyes revealed blanching of the ciliary processes at RF settings of 2.5 and higher with exposure times of 1.5 seconds or longer. Gross and light microscopic studies showed that levels of destruction correlated positively with RF settings and exposure times. Higher RF levels resulted in scleral coagulation necrosis on light microscopy. In addition, there was coagulation necrosis of the pigmented and nonpigmented ciliary epithelium and stroma. Longitudinal phase study showed a significant decrease in intraocular pressure (IOP) in the four groups of rabbits treated over the 4-week follow-up period (P = .005, multivariate analysis of variance [MANOVA]). Rabbits treated at higher RF levels sustained a greater IOP-lowering effect (P = .025, MANOVA). Gross and light microscopic examination revealed focal atrophy, fusion, and fibrosis of the ciliary processes. CONCLUSION: Nonpenetrating CTCD results in focal destruction of the ciliary body in rabbits. The authors found that significant reduction in IOP was possible over the 4-week follow-up period in the Dutch-belted rabbits treated, with greater IOP and tissue effects at higher RF settings. PMID- 8705746 TI - Experimental evaluation of a hydroxylapatite reservoir tube shunt in rabbits. AB - BACKGROUND AND OBJECTIVES: Anterior chamber tube shunts of various design are of increasing importance in the surgical management of refractory glaucoma. However, the reservoirs that are commonly used today are made of a silicone or an acrylic material that may become enveloped by a fibrous capsule, which in turn may impede aqueous resorption. In this study, the authors investigated the properties of hydroxylapatite (HA) as a reservoir in gonioimplants, using three different types of HA reservoir designs. It has been demonstrated that HA orbital implants become vascularized, and the authors hypothesized that this feature of HA might modify the development of the fibrous capsule, enhancing the efficacy and decreasing the size of the implants. MATERIALS AND METHODS: HA reservoirs with Silastic tube shunts were implanted in the right eyes of six Dutch-belted rabbits. Clinical examinations were performed and intraocular pressure was measured periodically for 16 weeks postoperatively. Prior to euthanasia, fluorescein-labeled latex microspheres were injected into the anterior chamber to demonstrate implant patency. Light microscopy was used to study the tissue response to the implants and the pathways of aqueous flow. RESULTS: In general, the authors found a granulomatous inflammatory response to the HA accompanied by neovascularization into the implant and minimal fibrous encapsulation of the reservoir. CONCLUSIONS: The use of HA as a suprascleral reservoir has several advantages, the major one being a theoretical increase in the functional lifetime of the implant. PMID- 8705747 TI - Monocular diplopia following excimer laser photorefractive keratectomy after radial keratotomy. AB - A 48-year-old man had symptoms of monocular diplopia following photorefractive keratectomy (PRK) for residual myopia after radial keratotomy. The cornea was characterized by a ring of subepithelial haze. A new type of corneal topography analysis that depicts areas of focal optical micro irregularity was used to diagnose the etiology of the optical symptoms. PMID- 8705748 TI - Malignant melanoma of the lacrimal sac. AB - The case of an 80-year-old woman who presented with a 1-year history of a right medial canthal mass and bloody tears and who was found to have a lacrimal sac melanoma is reported. A literature search revealed that only 16 cases of primary lacrimal sac melanoma have been reported. Current therapeutic modalities are discussed. PMID- 8705749 TI - HLA-B27--associated uveitis presenting with diffuse vitritis. AB - A 26-year-old man complained of a sudden decrease in vision in the left eye. He had a past medical history of pauciarticular juvenile rheumatoid arthritis with a positive HLA-B27 haplotype and negative antinuclear antibodies and later development of ankylosing spondylitis. His ocular history was unremarkable. An ophthalmic examination revealed a predominant diffuse vitritis (3+) in the left eye. The uveitis improved after treatment with topical and systemic steroids. Following an extensive clinical and laboratory evaluation, a final diagnosis of HLA-B27-associated uveitis was made. Although uncommon, diffuse vitritis can occur in the setting of this uveitis and may be a predominant symptom. PMID- 8705750 TI - A simple modification of the Hoskins lens for patients with dermatochalasis. AB - This brief report describes the use of transparent tape to modify the Hoskins lens for use in patients with dermatochalasis. The modification of the lens allows it to be used in these patients without the aid of an assistant. PMID- 8705751 TI - A fixation aid for slit-lamp biomicroscopic examination of the optic nerve. AB - The traditional articulated fixation light used on slit lamps is somewhat inconvenient, requiring repositioning for each eye. A simple, inexpensive fixation aid is described, which allows examination of both optic nerves without repositioning. PMID- 8705752 TI - Cracking of acrylic intraocular lenses during capsular bag insertion. PMID- 8705753 TI - Physical activity and the triggering of myocardial infarction: the case for regular exercise. PMID- 8705754 TI - Preservation of left ventricular function in mitral valve surgery. AB - Surgery for degenerative mitral regurgitation has become complex. Preservation of annulo-ventricular continuity through the chordae tendineae is an important determinant of operative survival, postoperative left ventricular function, long term survival, and quality of life. Some cardiologists believe that NYHA I function is never achieved after conventional mitral replacement with chordal transection. Valve repair is the procedure of choice but when valve replacement is inevitable every effort should be made to preserve the posterior leaflet and its chordal attachments. Valve replacement with preservation of the subvalvar apparatus provides a functional outcome similar to that after valve repair but usually leads to life long anti-coagulation. PMID- 8705755 TI - Cardioprotection: definition, classification, and fundamental principles. PMID- 8705756 TI - A British Cardiac Society survey of the potential for the secondary prevention of coronary disease: ASPIRE (Action on Secondary Prevention through Intervention to Reduce Events). AB - OBJECTIVE: To measure the potential for secondary prevention of coronary disease in the United Kingdom. DESIGN: Cross sectional survey of a representative sample of coronary patients from a retrospective review of hospital medical records and patient interview and examination. SETTING: Stratified random sample of 12 specialist cardiac centres and 12 district general hospitals drawn from 34 specialist cardiac centres and 261 district general hospitals in 12 geographic areas in the United Kingdom. SUBJECTS: 2583 patients < or = 70 yr; 25 consecutive males and 25 consecutive females identified retrospectively in each of four diagnostic categories: coronary artery bypass grafting, percutaneous transluminal coronary angioplasty, acute myocardial infarction, and acute myocardial ischaemia without evidence of infarction. MAIN OUTCOME MEASURES: Risk factor recording and management in medical records; the prevalence and control of risk factors at interview six months after the procedure or event. RESULTS: Recording of coronary risk factors in patient's records was incomplete and this varied by risk factor. Smoking habit and blood pressure were most completely recorded, whereas a history of hyperlipidaemia and blood cholesterol concentrations were least complete. Risk factor records were more likely to be complete in cardiac centres than in district hospitals. At interview 10% to 27% of patients were still smoking cigarettes and 75% remained overweight, females more severely so. Up to a quarter of patients remained hypertensive, males more severely so than females. Over three quarters had a total cholesterol > 5.2 mmol/l. In patients on medication for blood pressure, cholesterol or glucose, risk factor profiles were little better than in those who were not. Only about one patient in three was taking a beta blocker after infarction. Up to a fifth of patients who had had acute myocardial ischaemia were not taking aspirin at follow up. CONCLUSIONS: There is considerable potential to reduce the risk of a further major ischaemic event in patients with established coronary disease. This can be achieved by effective lifestyle intervention, the rigorous management of blood pressure and cholesterol, and the appropriate use of prophylactic drugs. PMID- 8705757 TI - Retained surgical swab misinterpreted as epicardial pacing wire on chest x ray. PMID- 8705758 TI - Ammonia response to exercise in patients with congestive heart failure. AB - OBJECTIVE: To assess energy depletion in skeletal muscle in patients with congestive heart failure by measuring blood purine metabolites during exercise and, at the same time, determine the implications of the ammonia response to exercise in these patients. SETTING: Tottori University Hospital, Yonago, Japan. PATIENTS: 49 heart failure patients (New York Heart Association (NYHA) grades I III) and 16 normal subjects. MAIN OUTCOME MEASURES: Blood lactate, ammonia, and hypoxanthine levels were measured during exercise with expired gas analysis. RESULTS: In normal exercising subjects as well as in each heart failure subgroup, the ammonia threshold was significantly higher than both the lactate threshold [control: 21.8 (SD 5.3) v 17.4 (3.3) ml/kg/min; NYHA class I: 18.9 (3.8) v 15.5 (2.6); class II: 14.8 (2.5) v 12.7 (2.4); class III: 13.5 (2.6) v 11.8 (2.5)] and the ventilatory threshold (P < 0.01). The difference between the ammonia and lactate thresholds was noted in all normal subjects and in all heart failure patients. The ammonia threshold, however, was significantly lower in heart failure patients than in normal subjects and it decreased with increasing NYHA class (P < 0.01). Maximum ammonia levels were lower in the heart failure group and decreased further with higher NYHA classifications [control: 198 (52) mg/dl; NYHA class I: 170 (74); class II: 134 (58); class III: 72 (15); P < 0.01]. There were significant correlations between maximum ammonia values and maximum lactate, oxygen consumption, and hypoxanthine levels (r = 0.74, 0.48, and 0.87, respectively; P < 0.001). CONCLUSIONS: The ammonia threshold may reflect the onset of ATP depletion in exercising skeletal muscles, as opposed to the onset of anaerobic respiration. It seems therefore that energy depletion in skeletal muscles during exercise occurs after attaining the anaerobic threshold. Both aerobic and anaerobic capacities of skeletal muscle are reduced in patients with congestive heart failure. PMID- 8705759 TI - Chronic intermittent haemofiltration and haemodialysis in end stage chronic heart failure with oedema refractory to high dose frusemide. AB - OBJECTIVE: To assess the benefits and problems of chronic intermittent treatment with haemofiltration or haemodialysis or both in patients with severe chronic heart failure (New York Heart Association class III or IV) and oedema refractory to pharmacological treatment. DESIGN AND SETTING: A retrospective case-cohort study. A general hospital in The Netherlands. PATIENTS: The results of chronic intermittent treatment with haemofiltration (n = 10) or haemodialysis (n = 2) were analysed in patients with severe chronic heart failure, predominantly due to coronary heart disease, and oedema refractory to a pharmacological regimen including high dose frusemide. INTERVENTION: Patients had an average of 25 (SD 38) treatments. RESULTS: There was improvement of NYHA class IV to III in seven patients. However, this was not reflected in a decrease in hospital admission: only two patients could be managed as outpatients. The median survival after start of the treatment was 24 days (varying from 0 to 393 days). In four patients the treatment was discontinued after discussion with the patient and family. CONCLUSIONS: The use of chronic intermittent haemofiltration and haemodialysis is of limited value in end stage chronic heart failure with oedema, refractory to maximal conventional treatment. PMID- 8705760 TI - Effects of gradual volume loading on left ventricular diastolic function in dogs: implications for the optimisation of cardiac output. AB - BACKGROUND: Volume loading is commonly used to adjust preload and optimise cardiac output. It is difficult to monitor preload at the bedside because filling affects ventricular diastolic function and consequently end diastolic pressure, which is the variable used to monitor preload. OBJECTIVE: To assess the effects of gradual volume loading on the different components of left ventricular diastolic function---filling velocities, relaxation, and chamber compliance---to identify how excessive loading produces diastolic dysfunction. METHODS AND RESULTS: Eight mongrel dogs, anaesthetised and mechanically ventilated with both the chest and the pericardium closed, were studied during basal conditions (B), during gradual volume loading with physiological saline---5 ml/kg (VL5), 10 ml/kg (VL10), and 15 ml/kg (VL15)---and during infusion of isosorbide dinitrate (10 g/kg/min) started after the VL15 load was achieved. Dogs were monitored haemodynamically and by transthoracic Doppler echocardiography to assess peak modal velocities of the E and A waves, E/A ratios, and the deceleration time of the E wave. M mode recordings of aligned mitral and aortic valve motion were also obtained to calculate the isovolumic relaxation time. Effects of volume loading on ventricular diastolic function seemed to occur in two phases. Small and moderate volume loads (VL5 and VL10) promoted early ventricular filling, increasing E wave velocities, improving the mean (SD) E/A ratio from 1.95 (0.3) (B) to 2.0 (0.27) (VL5) and 2.6 (0.3) (VL10) (P < 0.00005), prolonging the E wave deceleration time, and only slightly increasing ventricular diastolic pressures. These changes suggest an improvement in ventricular compliance. Extreme volume loads (VL15) produced an abrupt reduction in early ventricular filling, which was transfered to late in diastole, by decreasing E wave velocity, by increasing A wave velocity, and by decreasing E/A ratio from 2.6 (0.3) (VL10) to 0.8 (0.05) (VL15) (P < 0.00005). The E wave deceleration time was shortened and left ventricular diastolic pressures were much increased, all suggesting a deterioration in chamber compliance. All these restrictive changes were promptly reversed by the perfusion of isosorbide dinitrate. The isovolumic relaxation time steadily increased with volume loading. CONCLUSIONS: Small and moderate volume loads improved ventricular diastolic function by promoting early ventricular filling and increasing ventricular compliance. Extreme volume loads promptly induced a diastolic restrictive pattern, transferring filling to the second part of diastole (increasing dependence on atrial contraction) and reducing ventricular compliance. These changes in ventricular diastolic function were independent of simultaneously measured haemodynamic systolic performance and were promptly reversed by isosorbide dinitrate, which after extreme loading promoted early filling, myocardial relaxation, and improved chamber compliance. PMID- 8705761 TI - Matrix remodelling in dilated cardiomyopathy entails the occurrence of oncofetal fibronectin molecular variants. AB - OBJECTIVES: To investigate whether disturbance of the cellular homoeostasis and integrity of cardiomyocytes in dilated cardiomyopathy (DCM) is accompanied by alterations in cell-matrix relations as indicated by changes in the deposition of fibronectin (FN) isoforms. DESIGN: Tissue from a case series of patients with DCM was investigated by immunohistochemistry with antibodies against FN (all variants, clone IST4), ED-A+ FN (clone IST9), ED-B+ FN (clone BC1), and oncofetal glycosylated FN (clone 5C10). The sites of de novo synthesis of FN were demonstrated by means of non-radioactive RNA in situ hybridisation (ISH) with biotinylated FN cDNA fragments as the probe. SETTING: University hospital. PATIENTS: Samples from 10 patients with clinical criteria and histological diagnosis of DCM and from 3 individuals with normal hearts. INTERVENTIONS: Samples were obtained by right ventricular endomyocardial biopsy. MAIN OUTCOME MEASURE: Distribution of oncofetal FN variants in DCM hearts. RESULTS: Immunostaining of FN (IST4, all variants) showed a coarse interstitial network in normal and diseased myocardium. ED-A+ FN was deposited as fine interstitial spots in normal myocardium and in DCM samples. Immunostaining for oncofetal glycosylated FN and ED-B+ FN was not seen in normal adult myocardium, whereas myocardium from DCM patients showed focal and delicate staining in the interstitium. RNA ISH showed that these deposits resulted from local FN synthesis. CONCLUSION: The results accord with de novo expression of oncofetal FN variants in hearts from patients with DCM. The oncofetal FN variants may serve as disease markers in myocardium affected by DCM. PMID- 8705763 TI - Migration of an implantable cardioverter-defibrillator generator into the small bowel. PMID- 8705762 TI - Cardiac hypertrophy as a result of long-term thyroxine therapy and thyrotoxicosis. AB - OBJECTIVES: To define the effects of long-term thyroxine treatment upon heart rate, blood pressure, left ventricular systolic function, and left ventricular size, as well as indices of autonomic function, and to compare findings with those in patients with thyrotoxicosis before and during treatment. DESIGN: Cross sectional study of patients prescribed thyroxine long term (n = 11), patients with thyrotoxicosis studied at presentation (n = 23), compared with controls (n = 25); longitudinal study of patients with thyrotoxicosis studied at presentation and serially after beginning antithyroid drug treatment (n = 23). METHODS: 24 h ambulatory monitoring of pulse and blood pressure, echocardiography, forearm plethysmography, and autonomic function tests. RESULTS: Long-term thyroxine treatment in doses that reduced serum thyrotrophin to below normal had no effect on blood pressure, heart rate, left ventricular systolic function or stroke volume index, but was associated with an 18.4% increase in left ventricular mass index (mean (SEM) 101.9 (3.09) g/m2 v controls 86.1 (4.61), P < 0.01). Thryoxine treatment, like thyrotoxicosis, had no effect on tests of autonomic function. Untreated thyrotoxicosis resulted in pronounced changes in systolic and diastolic blood pressure and an increase in heart rate during waking and sleep. Patients with thyrotoxicosis at presentation had an increase in left ventricular systolic function (ejection fraction 70.5 (1.66)% v 65.4 (1.79), P < 0.01; fractional shortening 40.4 (1.54)% v 35.6 (1.46), P < 0.01), increased stroke volume index (45.9 (2.4) ml/m2 v 36.6 (1.7), P < 0.001), and an increase in forearm blood flow, and decrease in vascular resistance. They had a similar degree of left ventricular hypertrophy to that associated with thyroxine treatment (99.3 (4.03) g/m2); all changes were corrected within 2 months by antithyroid drugs. CONCLUSIONS: The development of left ventricular hypertrophy in patients receiving thyroxine in the absence of significant changes in heart rate, blood pressure, and left ventricular systolic function is consistent with a direct trophic effect of thyroid hormone on the myocardium. The presence of left ventricular hypertrophy determines that further studies are essential to assess cardiovascular risk in patients taking thyroxine long term. PMID- 8705764 TI - Coronary haemodynamics in left ventricular hypertrophy. AB - BACKGROUND: Left ventricular hypertrophy is associated with an increased risk of cardiovascular morbidity and mortality. Previous studies have shown that patients with left ventricular hypertrophy develop electrocardiographic changes and left ventricular dysfunction during acute hypotension, and suggest that the lower end of autoregulation may be shifted upwards. AIM: To measure coronary blood flow (velocity) and flow reserve during acute hypotension in patients with left ventricular hypertrophy. PATIENTS: Eight patients with atypical chest pain and seven with hypertensive left ventricular hypertrophy; all with angiographically normal epicardial vessels. SETTING: Tertiary referral centre. METHODS: The physiological range of blood pressure was determined by previous ambulatory monitoring. Left ventricular mass was determined by echocardiography. At cardiac catheterisation, left coronary blood flow velocity was measured using a Judkins style Doppler tipped catheter. During acute hypotension with sodium nitroprusside, coronary blood flow velocity was recorded at rest and during maximal hyperaemia induced by intracoronary injection of adenosine. Quantitative coronary angiography was performed manually. RESULTS: For both groups coronary blood flow velocity remained relatively constant over a range of physiological diastolic blood pressures and showed a steep relation with diastolic blood pressure during maximal hyperaemia with intracoronary adenosine. Absolute coronary blood flow (calculated from quantitative angiographic data), standardised for left ventricular mass, showed reduced flow in the hypertensive group at rest and during maximal vasodilatation. CONCLUSION: The results are consistent with an inadequate blood supply to the hypertrophied heart, but no upward shift of the lower end of the autoregulatory range was observed. PMID- 8705765 TI - Relation between symptoms and profiles of coronary artery blood flow velocities in patients with aortic valve stenosis: a study using transoesophageal Doppler echocardiography. AB - OBJECTIVE: To analyse profiles of coronary artery flow velocity at rest in patients with aortic stenosis and to determine whether changes of the coronary artery flow velocities are related to symptoms in patients with aortic stenosis. DESIGN: A prospective study investigating the significance of aortic valve area, pressure gradient across the aortic valve, systolic left ventricular wall stress index, ejection fraction, and left ventricular mass index in the coronary flow velocity profile of aortic stenosis; and comparing flow velocity profiles between symptomatic and asymptomatic patients with aortic stenosis using transoesophageal Doppler echocardiography to obtain coronary artery flow velocities of the left anterior descending coronary artery. SETTING: Tertiary referral cardiac centre. PATIENTS: Fifty eight patients with aortic stenosis and 15 controls with normal coronary arteries. RESULTS: Adequate recordings of the profile of coronary artery flow velocities were obtained in 46 patients (79%). Left ventricular wall stress was the only significant haemodynamic variable for determining peak systolic velocity (r = -0.83, F = 88.5, P < 0.001). The pressure gradient across the aortic valve was the only contributor for explaining peak diastolic velocity (r = 0.56, F = 20.9, P < 0.001). Controls and asymptomatic patients with aortic stenosis (n = 12) did not differ for peak systolic velocity [32.8 (SEM 9.7) v 27.0 (8.7) cm/s, NS] and peak diastolic velocity [58.3 (18.7) v 61.9 (13.5) cm/s, NS]. In contrast, patients with angina (n = 12) or syncope (n = 8) had lower peak systolic velocities and higher peak diastolic velocities than asymptomatic patients (P < 0.01). Peak systolic and diastolic velocities were -7.7 (22.5) cm/s and 81.7 (17.6) cm/s for patients with angina, and -19.5 (22.3) cm/s and 94.0 (20.9) cm/s for patients with syncope. Asymptomatic patients and patients with dyspnoea (n = 14) did not differ. CONCLUSIONS: Increased pressure gradient across the aortic valve and enhanced systolic wall stress result in characteristic changes of the profile of coronary flow velocities in patients with aortic stenosis. Decreased or reversed systolic flow velocities are compensated by enhanced diastolic flow velocities, particularly in patients with angina and syncope. This characteristic pattern of the profile of coronary artery flow velocities in patients with angina or syncope may be useful for differentiating those patients from asymptomatic patients. PMID- 8705766 TI - Pulmonary oedema and pleural effusion in two patients with primary pulmonary hypertension treated with calcium channel blockers. PMID- 8705767 TI - Catheter ablation for successful management of left posterior fascicular tachycardia: an approach guided by recording of fascicular potentials. AB - OBJECTIVE: To assess whether catheter ablation of fascicular tachycardia can be facilitated by the recording of sharp deflections arising from the mid-septum-- inferior apical septum of the left ventricle. PATIENTS AND METHODS: Seven consecutive patients (mean age 29 (range 16-43) years) with ventricular tachycardia originating from the left posterior fascicle underwent electrophysiology study and detailed mapping of endocardial activation. Selection of ablation sites in the last five patients was based on the recording, during left posterior fascicular tachycardia and sinus rhythm, of a discrete potential preceding the earliest ventricular electrogram, which was thought to represent conduction through the posterior fascicle. RESULTS: Patients were treated with low energy direct current or radiofrequency current ablation. The median fluoroscopy and procedure times were 23 (range 6-42) min and 110 (range 50-176) min, respectively. In a follow up period of 4 to 16 months, six patients were asymptomatic and one had minor symptoms. No patient had any change in intraventricular conduction. Similar potentials were also recorded from the left posterobasal septum in three of eight patients who underwent catheter ablation of left free wall accessory pathways. CONCLUSION: Fascicular potentials can be reproducibly recorded in left posterior fascicular tachycardia and may serve as a reliable marker for successful ablation procedures. The relation of these potentials with the substrate of the tachycardia, however, remains obscure. PMID- 8705768 TI - Aortic root complications in Marfan's syndrome: identification of a lower risk group. AB - OBJECTIVES: The purpose of this study was to examine clinical and echocardiographic predictors of outcome in a cohort of patients with Marfan's syndrome. BACKGROUND: Serial echocardiographic measurements of aortic root dimensions are an important clinical method for monitoring patients with Marfan's syndrome. However, there are few data on the prognostic importance of echocardiographic variables for risk stratification and timing of aortic root replacement. METHODS: In 89 consecutive patients with Marfan's syndrome (age range 1-54 years) clinical and serial echocardiographic data (n = 62) were evaluated as potential predictors of outcome (mean (range) follow up 4 (< 1-16) years). Aortic sinus diameter and an aortic ratio normalised for age and body surface area were examined using Kaplan-Meier life table and Cox regression analysis, with the end point defined as death or surgery for ascending aortic dissection and events defined as an end point or surgery for ascending aortic aneurysm. RESULTS: Overall actuarial survival at two and five years was 96% and 92% and event free survival was 85% and 76%, respectively. There were five deaths due to aortic dissection, four patients survived surgery for ascending dissection, and nine underwent root replacement with a composite graft for ascending aneurysm. Those with aortic events were older (35 (12) v 25 (13) years, P = 0.007) and had greater initial aortic root dimensions (47 (14) v 33 (8) mm, P < 0.0001) and ratios (1.6 (0.5) v 1.3 (0.2), P < 0.0001). In the 62 patients with serial echocardiographic follow up, the rate of aortic root dilatation was more rapid in those with events (15 (17) v 0 (3)%/year, P < 0.0001). Utilising a Cox proportional hazards model, the groups with an initial aortic ratio > or = 1.3 or an annual change in aortic ratio > or = 5% had a relative risk of an aortic complication of 2.7 and 4.1, respectively (95% confidence limits 1.5 to 4.8 and 1.8 to 9.3). Only one of 31 patients with an initial aortic ratio of < 1.3 and a rate of change of < 5% had an event (five year event free survival 97%). CONCLUSIONS: A low risk subgroup of patients with Marfan's syndrome can be identified as those with an aortic ratio < 1.3 and an annual change in aortic ratio of < 5%. These findings are helpful in optimising echocardiographic monitoring and risk stratification of patients with Marfan's syndrome. PMID- 8705769 TI - Prognosis of supravalve aortic stenosis in 81 patients in Liverpool (1960-1993). AB - OBJECTIVE: To determine the prognosis of supravalve aortic stenosis into early adult life and the factors affecting this prognosis. DESIGN: 81 patients with supravalve aortic stenosis were followed for a median duration of 8.3 (range 1 to 29) years. PATIENTS: 40 patients (49.4%) had Williams' syndrome, 18 (22.2%) familial supravalve aortic stenosis, 18 (22.2%) sporadic supravalve aortic stenosis, and five (6.2%) other syndromes. Nineteen patients had additional levels of left ventricular outflow tract obstruction. RESULTS: 47 patients (58%) underwent operation; 20% within a year of presentation. Multivariable analysis predicted that 88% of patients would undergo intervention within 30 years of follow up. The chance of intervention was increased by more severe aortic stenosis at presentation and the presence of multilevel obstruction in patients with sporadic supravalve aortic stenosis. Three deaths occurred before operation and 13 within a month of operation. Ten (62.5%) of the postoperative deaths were in patients with multilevel obstruction. Predicted survival 30 years after presentation was 66%. Risk factors for survival were age and severity of aortic stenosis at presentation. Multilevel obstruction did not emerge as a significant risk factor for death because of the high association with the severity of stenosis at presentation. 74% of survivors had mild or insignificant stenosis at follow up. CONCLUSIONS: Long-term survival is related to age and the severity of aortic stenosis at presentation. Most patients will require intervention, and most survivors will have mild stenosis. PMID- 8705770 TI - Surgical preconditioning and completion of total cavopulmonary connection by interventional cardiac catheterisation: a new concept. AB - A new, staged procedure for univentricular repair of "high risk" patients using a combined surgical transcatheter technique is reported. As first step a surgical hemifontan procedure was performed and a multifenestrated patch inserted into the right atrium. For later transcatheter completion of univentricular repair, a subtotal banding just above the cavoatrial junction was performed. Two months later transcatheter completion was performed by balloon dilatation of the banded cavo-atrial junction and additionally either by inserting a covered stent as intracardiac conduit between the superior and inferior vena cava or by inserting a (non-covered) stent into the cavo-atrial junction and occluding the fenestrations in the right atrial patch using Rashkind-PDA occluders. This new technique was successfully applied without mortality in eight patients with a preoperative mean pulmonary pressure of 18 to 23 mm Hg. No complications typical for Fontan-like corrections occurred within the follow up period of 4 to 14 months. PMID- 8705771 TI - Stability of plasma concentrations of N and C terminal atrial natriuretic peptides at room temperature. AB - BACKGROUND: Plasma concentrations of atrial natriuretic peptide (ANP) are increased in patients with ventricular dysfunction and could have a diagnostic role in heart failure. ANP may be unstable after collection, however, limiting any practical diagnostic role. METHODS: Blood samples were obtained from 18 patients with various conditions. Aliquots were either processed optimally or kept as blood or plasma at room temperature for 6-72 h before processing. RESULTS: Concentrations of C-terminal ANP were lower in specimens kept as blood for 24 and 72 h (mean difference from control -43% and -76%, respectively, (P < 0.001) but N-terminal ANP (extracted) seemed to be stable under all conditions studied (-2% at 24 h and -7% at 72 h, not significant). CONCLUSIONS: N-terminal ANP (extracted) is stable and potentially has a role in the diagnosis of heart failure in routine clinical practice. PMID- 8705772 TI - Persistent left sided and absent right sided superior vena cava complicating permanent pacemaker insertion. PMID- 8705773 TI - Shuttle walking test: a new approach for evaluating patients with pacemakers. AB - OBJECTIVE: To validate an incremental field walking test, the shuttle walking test, as a means of assessing pacing modes and to aid programming of rate responsive pacemakers. DESIGN: Three separate groups of patients were recruited. Reproducibility (n = 10) of the shuttle walking test was assessed by performing three consecutive tests. Comparison of the shuttle walking test with a 10 min walk was assessed in 20 patients. In the third group (n = 10) patients with rate responsive pacemakers were programmed to either VVI fixed rate 70 beats/min or VVIR with the optimal rate response to show the discriminative value of the test. SETTING: Pacing clinic in a regional cardiothoracic centre. PATIENTS: 30 patients with chronotropic competence and dual chamber pacemakers with varying functional capacity and 10 patients with rate responsive pacemakers. INTERVENTIONS: Continuous haemodynamic monitoring was obtained using an ambulatory nuclear monitor, the Capintec VEST. Two exercise tests either shuttle walking test or 10 min corridor walk. The shuttle walk is an incremental walking test conducted on a 10 m course where the walking speed is dictated by bleeps on an audio cassette. RESULTS: Reproducibility was demonstrated over three consecutive tests with mean (1 SD) exercise times of 7.6 (1.7) min, 7.7 (1.6), and 7.7 (1.7) min. During the shuttle walk the test patients walked for a mean of 8.3 (1.2) min producing peak relative cardiac outputs of 78 (21) end diastolic volume/min compared with 64.9 (17) end diastolic volume/min for the 10 min walk (P < 0.001); peak heart rates were 118 and 104 beats/min (P < 0.03) respectively. In the third group relative peak cardiac output was significantly greater in VVIR (70 (24) v VVI 52 (15) end diastolic volume/min) (P < 0.009) as were exercise times (VVIR 8.8 (1.3) min v VVI 8.1 (1.3) min) (P < 0.003). CONCLUSIONS: The shuttle walk is an easy test to administer, requiring little equipment. It produces a symptom limited maximal performance and will be a useful aid to pacemaker programming as it is reproducible and able to show differences in exercise capacity between pacing modes. PMID- 8705774 TI - Planning for coronary angioplasty: guidelines for training and continuing competence. British Cardiac Society (BCS) and British Cardiovascular Intervention Society (BCIS) working group on interventional cardiology. AB - The following recommendations are made: 1 Existing centres undertaking angioplasty should increase their activity, and the target figure of 400 PTCA procedures per million of the United Kingdom population should be achieved by the end of 1996-97, or immediately thereafter. 2 Angioplasty centres should be appropriately equipped to undertake PTCA safely and effectively and provide a reliable emergency service. They should have a minimum of two trained PTCA operators jointly undertaking a minimum of 200 procedures per year at that centre, and have regular meetings to share experience. 3 Angioplasty operators should ensure that where the need arises patients undergoing PTCA can receive immediate attention from a trained operator at any time until discharge from hospital. 4 Trained operators should undertake at least 1-2 PTCA procedures per week (> 60 procedures per year) to maintain competence, and those undertaking so few procedures should increase their activity over the next three years to more than 100 a year. 5 Trainers should have performed at least 500 procedures before formally training others and should undertake a minimum of 125 procedures a year to maintain accreditation as a trainer. 6 Surgical cover for PTCA procedures should be mandatory and on site cover remains the strongly preferred option. Where surgical cover is provided off site, this should be at a centre less than 30 minutes away by road. Whether provided on or off-site it should be possible to establish cardiopulmonary bypass within 90 minutes of the decision being made to refer the patient for surgery. 7 All operators and interventional centres should audit their activity and results, review these data locally with colleagues, and provide regular audit returns to the national database run by BCIS. This will allow future recommendations concerning standards to take more account of risk stratification and actual outcomes, and not place such emphasis merely on volumes of activity. 8 These recommendations should be reviewed in three years. PMID- 8705775 TI - Atrial thrombi occurring during sinus rhythm in cardiac amyloidosis: evidence for atrial electromechanical dissociation. PMID- 8705776 TI - Is aspirin safe in heart failure? more data. PMID- 8705777 TI - New directions in anticoagulant and antiplatelet treatment. PMID- 8705778 TI - Lingual haematoma after treatment with alteplase (recombinant tissue plasminogen activator) for acute myocardial infarction. PMID- 8705779 TI - Misdiagnosis of lung cancer in a 2000 consecutive autopsy study in Budapest. AB - OBJECTIVE: To determine the accuracy of lung cancer mortality data based on clinical observations in the absence of autopsy and to identify factors affecting the accuracy of diagnosis. METHODS: Admission, pre-autopsy and post-autopsy diagnoses were recorded for 1000 consecutive autopsies in each of two University departments in Budapest with high autopsy rates for persons dying in hospital. In those 87 cases where one or more diagnosis included primary lung cancer, additional data were collected concerning clinical investigations relevant to the diagnosis and the histological type lung cancer, and on smoking habits. RESULTS: 59% (36/61) of lung cancers seen at autopsy were not detected pre-autopsy, while 50% (25/50) of those diagnosed pre-autopsy were not confirmed at autopsy. Many misdiagnoses arose because patients were too ill to be properly investigated and/or died before investigations could be completed. Accuracy of diagnosis increased with the number of diagnostic techniques applied, but was still far from perfect in the absence of necropsy. Underdiagnosis was commoner in non smokers and overdiagnosis commoner in smokers. CONCLUSIONS: Without necropsy, lung cancer misdiagnosis is common, especially when modern diagnostic procedures cannot be fully employed. Knowledge of smoking habits may affect diagnostic accuracy. PMID- 8705780 TI - Analysis of the hepatocyte growth factor receptor in regeneration and oncogenesis of the liver. AB - The c-met proto-oncogene product is the tyrosine kinase receptor for hepatocyte growth factor. To gain insights into its functions in the liver, we carried out a study of the expression and tyrosine phosphorylation status of the Met protein during hepatic regeneration and in human hepatocellular carcinomas. Following partial hepatectomy of rats, decreased expression of the proreceptor p170MET and reduced levels of tyrosine phosphorylation were seen within 12 h. These changes were still evident 7 days later. By contrast, there was no significant alteration of the Met beta subunit. The analysis of samples from 18 patients with hepatocellular carcinoma revealed that the expression of p160MET proreceptor was increased in non-cancerous areas, while that of the p145 beta MET proreceptor was significantly greater in the tumor tissue than in the non-tumor areas (p < 0.05). These findings suggest that processing of the Met proreceptor is closely associated with regeneration and carcinogenesis of the liver. PMID- 8705781 TI - Electron microscopic investigation on Ito cells and Disse's space in patients with extrahepatic cholestasis. Immunohistochemistry of collagen type IV and fibronectin in hepatic sinusoids. AB - Extrahepatic cholestasis is one of the main factors causing liver fibrosis. Surgical biopsies of six patents were studied: one with normal liver (control patient), and five with different degrees of extrahepatal cholestasis with hepatocellular degenerative and necrotic changes. Monospecific antibodies directed against type IV collagen and fibronectin were localized by light microscopical immunohistochemistry. Type IV collagen was found in all basement membranes: ductal, vascular, neural, Intensive, almost continuous deposits of it were found along the entire length of the sinusoid. Fibronectin, the structural glycoprotein, was codistributed with type IV collagen in portal matrix and in perisinusoidal location. It did not decorate the basement membranes of bile ducts and stained more intensely than type IV collagen at the border between portal stroma and parenchyma. The intensity of the two antibodies increased corresponding with the heaviness of parenchymal deterioration. There was weaker staining behavior of the extracellular matrix regarding collagen IV and fibronectin in the area of heavy bilirubin impregnation. The sinusoid morphology in the regions with fibrosis and increased extracellular matrix formation in periportal areas was reported. Disses's space was distended and occupied by collagen bundles, amorphous matrix, swollen hepatocyte microvilli, and basement membrane-like material. Ito cells transformed into transitional cells or myofibroblast-like cells. Sinusoidal changes and increased collagen IV as well as perisinusoidal fibronectin formation coincided with the aggravation of cholestasis. PMID- 8705782 TI - On the resorcin-fuchsin stainings. III. Demonstration of sulfated glycoproteins and proteoglycans. PMID- 8705783 TI - EGF-R and PCNA expression in ovarian carcinomas--correlation with classic prognostic factors. AB - Expression of the epidermal growth factor-receptor (EGF-R) and the proliferating cell nuclear antigen (PCNA) was immunohistochemically studied in 75 ovarian cancer samples using formalin-fixed, parafin-embedded tissue. Correlations between these factors and conventional histomorphologic factors were investigated. 44 (58.7%) tumors were EGF-R positive (> 10% positive cells). 18 tumors (24%) showed a weak EGF-R-expression (< or = 50% positivity) and 26 tumors (34.7%) had a strong EGF-R-expression. Expression of EGF-R did not correlate with any of the other prognostic factors investigated. The PCNA-proliferative fraction was classified using the median value (< or = 34%/ > 34%) and a categorization in three groups (< 20%/20%-50%/ > 50%). PCNA-expression showed no correlation with FIGO-stage, histologic tumor type, lymph node-status and EGF-R. However, both PCNA-classifications correlated with the size of the residual tumor (PCNA < or = 34%/ > 34%/p = 0.046; PCNA < 20%/20%-50%/ > 50%/p = 0.086) and the histologic grading (p = 0.076; p = 0.02 respectively). Thus, the PCNA-proliferative fraction might be an additional indicator for tumor aggressiveness and disease outcome. PMID- 8705784 TI - Adrenocortical nodules in post-mortem series. Development, functional significance, and differentiation from adenomas. AB - In 498 consecutive autopsy cases, the adrenal glands were light microscopically and morphometrically studied. 265 (53.7%) patients showed single or multiples nodules, 25 (5.0%) adenomas. The diameter of nodules was between 0.3 to 8.0 mm and that of adenomas between 3.2 to 28.0 mm. Clinically, 265 (36.5%) patients revealed arterial hypertension. 283 (63.5%) were normotensive. For the diagnosis of adenomas, the architecture and the cell structure were more important than the size of the tumor. Normotensive patients showed more often (55.%) nodules than hypertensive patients (52.8%). Adenomas were more frequently found in hypertensive patients. There was no correlation between age, sex and nodules, but adenomas were more frequently found in females. Normotensive patients exhibited mainly one nodule, whereas multiple nodules were found in hypertensive patients. In patients with nodules arteriosclerosis with intima hyalinosis was present in 184 (56.3%) cases, in patients without nodules in 81 (47.4%) cases. Our study clearly demonstrates that no correlation exists between the occurrence of adrenocortical nodules and age or hypertension. The etiology of adrenal nodules should be considered together with the arteriopathy. PMID- 8705785 TI - Diffuse form of idiopathic IgA nephropathy (IgAN). A quantitative study. AB - Fifteen renal biopsy specimens from patients with idiopathic diffuse IgAN for whom both light and electron microscopy as well as immunofluorescence microscopy and full clinical data were available were quantitatively examined and compared with fifteen cases of normal controls. Morphometric investigations were performed by means of a computer image analysis system to evaluate whether morphometric analysis may be helpful in morphologic diagnosis of IgAN and to study whether serum creatinine and changes in quantitatively analyzed glomeruli could correlate with the interstitial fibrosis. The study revealed that total glomerular area, total number of glomerular cells per total glomerular area, and total glomerular cells per unit of glomerular area as well as relative interstitial volume were increased in IgAN in comparison with normal glomeruli, most of them significantly. Moreover, there were significant positive correlations between the width of the cortical interstitium and serum creatinine level as well as between the width of the cortical interstitium and glomerular changes. Although these findings suggest that in IgAN both glomerular and interstitial lesions are involved in the renal prognosis, the authors conclude that a morphometric analysis does not lend itself to establish such causal associations. It was noted that morphometric methods supported by image analysis system cannot be used instead of light microscopy, electron microscopy, and immunohistochemistry, but they can be helpful in precise evaluation of both glomerular and interstitial lesions, which are commonly known as histologic risk factors in primary glomerular diseases. PMID- 8705786 TI - A technique for automatic/interactive assessment of the proliferating fraction of neoplastic cells in solid tumors. A methodological study on the Ki-67 immunoreactive cells in human mammary carcinomas, including a comparison with the results of conventional S-phase fraction assessments by means of DNA cytometry. AB - Based on a computerized microscopy technique, a method has been devised which allows the practising pathologist to easily and rapidly assess quantitatively the relative number of actively proliferating neoplastic parenchymal cells in a tumor nodule. Our method has been tested on a series of 20 conventionally formalin fixed and paraffin-embedded female mammary adenocarcinomas, using immunoreactivity with the MIB-1 monoclonal antibody against the cell proliferation antigen Ki-67. The values of the proportion of the MIB-1 immunoreactive cell nuclei were compared with those obtained DNA-cytometrically for the fraction of cells in the S-phase; a good correlation was found, although the MIB-1 values were consistently somewhat higher. A prerequisite for a success of the method was, of course, to achieve standardization of the MIB-1 immunostaining technique. By making simple adjustments of it, it could actually be improved to such an extent that almost the same color calibration and thresholding setup could be used. The measuring technique could be either interactive or automatic. The total number of immunoreactive and non immunoreactive nuclei, as well as the total nuclear area of both cell types were registered in a computerized device. The data were accumulated sequentially for each measure field. To investigate the reproducibility of the immunostaining, two slides of each case were stained on different occasions. Each slide was measured three times; systemically randomly in the x- and y-axis-directions as well as in the subjectively defined histopathologically "most proliferative" area of the tumor. The values obtained were in good agreement with each other and obviously gave some valuable and objective supplementary pieces of information to that of the conventional clinical and histopathologic assessment of the degree of aggressiveness of a malignant neoplasm. PMID- 8705787 TI - Radioimmunological and immunohistochemical study of carcinoembryonic antigen in pleomorphic adenoma and mucoepidermoid carcinoma of the salivary glands. AB - The aim of the present study was the evaluation of CEA-radio-immunoassay and CEA immunohistochemistry in the management of pleomorphic adenoma and mucoepidermoid carcinoma of salivary glands. 23 pleomorphic adenomas, 9 mucoepidermoid carcinomas, and 7 normal salivary glands were examined. CEA-concentration in serum and saliva were assayed before and after surgery. Polyclonal CEA antibody was used for immunohistochemical CEA detection in the tumor tissue and in the normal salivary glands. The mean CEA concentrations were found to be 14.94 ng/ml in the serum and 216.67 ng/ml in the saliva of patients with mucoepidermoid carcinoma. These values were considerably higher compared to healthy controls (188.64 ng/ml in saliva) and in patients with pleomorphic adenoma - 7.65 ng/ml in serum and 189.35 ng/ml in saliva (P < 0.001). A correlation was found between high CEA concentration in the saliva and the intensity of CEA expression in the tumour tissue. An increased synthesis and secretion of CEA was determined by the prevalence of tubular structures, a high proliferative activity in pleomorphic adenoma, and its malignant transformation. PMID- 8705788 TI - Primary non-Hodgkin lymphomas of the CNS-proliferation, oncoproteins and Epstein Barr-virus. AB - Primary cerebral lymphomas (PCL's) are rare tumors which, however, occur with increasing frequency. The present study investigated 55 PCL's of B-cell type, 36 in immunocompetent and 19 in AIDS-patients and 6 cases of intravascular lymphomatosis. In immunocompetent patients, proliferative indices as evaluated by PC10 and MIB1 reflected the histologic grade. Low grade tumors had a mean PCNA and MIB-1 count of 19 and 18.8 (SD 14.7 and 13.2), respectively, and high grade neoplasias showed counts of 56.7 and 47.1 (SD 19 and 17.4), respectively. No correlation of both indices with patient survival was found. 21 cases (58.3%) displayed p53-positivity of varying degree and 19 cases (52.7%) harbored bcl-2 positive neoplastic cells. Immunocompetent cases were always negative for Epstein Barr virus RNA and lmp-1-protein. In AIDS-cases, 13 cases (68.4%) showed up lmp-1 positivity and 15 cases (78.9%) had EBER-RNA. bcl-2 positive cells were detected in 5 cases (26.3%) and all cases were p53-negative. These results are in keeping with a role of EBV in the pathogenesis of primary cerebral lymphomas in AIDS-, but not in immunocompetent patients. None of the cases with intravascular lymphomatosis showed an expression of bcl-2 or p53 oncoproteins or lmp-1 and none had EBER-RNA. PMID- 8705789 TI - DNA cytometry in renal cell carcinoma. AB - The course of patients suffering from renal cell carcinoma depends on various factors. The measurement of the nuclear DNA content has been considered as a prognostic indicator in addition to morphologic and clinical parameters. The meaning of this parameter is under discussion today. The aim of the present study is to investigate the influence of nuclear DNA parameters of renal cell carcinoma on the prognosis. It should be searched for such parameters which give us the possibility to divide the patients into a low-risk-group and a high-risk-group. 110 clear cell renal carcinomas were investigated by means of image analysis and flow cytometry. Stemline ploidy, 5c-exceeding-rate, DNA grade of malignancy, S phase-fraction, proliferating fraction, and nuclear area have been determined. A significant correlation between the size of the tumor, lymph node metastases and distant metastases on the one hand and the computed DNA parameters on the other hand was found. A similar correlation between histologic grading and DNA parameters could be found. There were pieces of information about the clinical follow-up in 87 patients (mean follow-up time 15,5 months). Local recurrence occurred in 12 cases. The computed differences between DNA parameters and local recurrence were not significant. Further clinical follow-up and clinical re examination of all patients will be performed. Then we can decide, if it is possible to make a division into low-risk and high-risk cases based on DNA parameters. PMID- 8705790 TI - Cytomorphologic findings in fine needle aspiration specimens of eosinophilic renal cell carcinoma. AB - The cytologic morphology of an eosinophilic renal cell carcinoma verified by histology is demonstrated. We describe the morphologic criteria for the differential diagnosis of the types of renal cell carcinoma in fine needle aspiration cytology and discuss the role of this procedure in the diagnosis of the entity. PMID- 8705791 TI - Perforation of the small intestine in a patient with disseminated Pneumocystis carinii infection in AIDS. AB - Perforation of the small intestine due to a segmental transmural Pneumocystis carinii infiltrate of the whole circumference was found in a surgical resection specimen as the cause of an acute abdomen in a 48-year-old heterosexual male patient suffering from acquired immunodeficiency syndrome. On autopsy, a disseminated Pneumocystis carinii infection was found involving spleen, thyroid gland and lymph nodes. The origin of this disseminated infection was a recurrent and severe Pneumocystis carinii pneumonia, which was first diagnosed two years before death and was treated with success. The hitherto unknown complication of an extrapulmonary Pneumocystis carinii infection described here extends the spectrum of lethal complications of opportunistic infections in acquired immunodeficiency syndrome. PMID- 8705792 TI - Clear cell meningioma: report of a spinal case. AB - A sacrococcygeal case of clear cell meningioma in a 38-year-old man is reported. Compared to the seven spinal clear cell meningiomas described so far, this case demonstrated the highest recurrency rate. Multiple recurrences were most likely due to the extensive infiltrative growth pattern of the tumor hindering complete microscopic surgical resection. Histologic parameters were not relevant to predict recurrences, since cellular anaplasia was lacking and growth fraction was low in the first operation and all subsequent recurrences. In view of what is now known of clear cell meningiomas, the tumor showed all histologic and immunohistochemical features which are typical for this tumor entity. However, the tumor was first diagnosed by capable pathologists and neuropathologists as fibroma-like mesenchymal tumor, metastatic renal carcinoma, chordoma, chondroma untypical osteosarcoma, and microcystic meningioma. Correct diagnosis was made by electron microscopy which has revealed comparable findings to those of other rare electron microscopical cases reported so far; high content of cytoplasmic glycogen diffusely distributed throughout the cytoplasm or aggregated within vacuoles, intermediate filaments, desmosomes, interdigitation of cell membranes and large amounts of collagen fibers, some of which were of the giant amianthoid type. Since meningiomatous features of clear cell meningiomas are not obvious light microscopically and the tumors may be confused with nonmeningiomatous neoplasms, electron microscopical investigation is highly recommended in each case of suspected clear cell meningioma. PMID- 8705793 TI - Tonsillar metastases: report on two cases and review of literature. AB - Metastases to the tonsils are extremely infrequent. Less than 70 cases have been reported in literature since 1858. The commonest sources of tonsillar metastases are malignant melanomas and carcinomas of the breast and the lungs. We report about two new cases of tonsillar metastases, one of which developed from a malignant melanoma, the other one from a hepatocellular carcinoma. We have not found any reports on tonsillar metastases stemming from hepatocellular carcinomas in literature and, moreover, in our case, this was the clinical presentation of the primary tumor. PMID- 8705794 TI - Intracortical low grade osteosarcoma. A unique case and review of the literature on intracortical osteosarcoma. AB - The thirteenth case of an intracortical osteosarcoma is presented. According to the authors' knowledge, this case appears to be the first report of an intracortical low grade osteosarcoma of the fibrous dysplasia-like variant in the femur. All other cases (except one small cell variant) reported in the literature were histologically diagnosed as high grade osteoblastic or sclerotic variants. The finding of both a low grade and a small cell variant of osteosarcoma supports the concept that intracortical osteosarcoma rather represents a distinct entity defined by location than an early detected conventional medullary osteosarcoma. PMID- 8705795 TI - Hepar lobatum carcinomatosum due to chemotherapy of a metastatic breast carcinoma. AB - Besides the lungs, the liver is the second most common site of hematogenous metastases from carcinomata of the breast. Hepar lobatum carcinomatosum is the rarest form of metastatic liver disease. Reported in this article is a case of a 59-year-old woman with invasive duct carcinoma of the breast with metastasis to the axillar lymph nodes and liver, treated with ablatio mammae and combination of chemotherapy. The etiology of hepar lobatum is caused by multiple pathogenetic factors. Tumor-related multifocal obstruction of portal and hepatic venous vessels and effects of chemotherapy are discussed. PMID- 8705796 TI - Decision making in ruminant orthopedics. AB - Decision making in ruminant orthopedics is determined by many factors, the most of important of which is age, size, and value of the patient, the nature of the injury, the prognosis for effective treatment and satisfactory healing, the intentions of the client, and the experiences of the veterinarian. Ruminant orthopedics currently is expanding to include the treatment of llamas and small ruminants as companion animals in addition to the treatment of valuable livestock. The future promises increasing sophistication in treatments and an ever higher quality of patient care. PMID- 8705797 TI - External skeletal fixation in ruminants. AB - External skeletal fixation (ESF) techniques are increasingly being adapted for treatment of fractures in large animals. This article includes detailed information regarding decision making for ESF, biomechanics of ESF, interaction of the bone and implants, general surgical techniques for application of ESF (including traditional ESF, use of acrylic sidebars for ESF, and transfixation, pinning, and casting), discussions of application of ESF to specific bones, and complications associated with the use of ESF. Also, techniques for the management of open, infected fractures are presented. Future directions of ESF research and application also are presented. PMID- 8705798 TI - Fractures of the humerus. AB - The treatment of humeral fractures in ruminants remains a challenge to the large animal surgeon. Often the recommendation is to euthanize the animal. This article reviews four different treatment techniques: stall confinement, intramedullary pinning, interlocking nails, and plate fixation. Clinical cases are drawn from both the literature and personal experience to demonstrate the potential application of each treatment modality. PMID- 8705799 TI - Fractures of the femur. AB - Fractures of the femur in cattle are second in frequency only to fractures to the metacarpal and metatarsal bones. Common clinical presentations and the diagnosis of femur fractures are described. Treatment options, prognosis, postoperative complications are discussed for conservative therapy and for various forms of external and internal fixation. PMID- 8705800 TI - Treatment of fractures of the tibia and radius-ulna by external coaptation. AB - External coaptation of radial-ulnar and tibial fractures with casts or modified Thomas splint-cast combinations is a useful treatment. The economics of therapy make this method of treatment feasible for commercial animals. Current estimates for the cost of treatment of tibial fractures with Thomas splint-cast combinations are $225.00 for calves and $410.00 for cattle if the metal splints are reused. Casts have similar costs. The availability of external coaptation techniques to all veterinarians and the success of treatment make external coaptation a good method for the treatment of many tibial and radial-ulnar fractures in cattle. PMID- 8705801 TI - Fracture biology, biomechanics, and internal fixation. AB - The success of orthopedic surgery in ruminants is directly related to the surgeon's knowledge and understanding of bone physiology and mechanics. The relationship of the macro and micro structure and function of bone as it relates to fracture physiology and repair is discussed. A basic review of the biomechanical principles of bone, bone fracture, and fracture repair are presented. The clinical and biomechanical principles of internal fixation are described for fracture repair in ruminants. PMID- 8705802 TI - Metacarpal and metatarsal fractures in cattle. AB - Metacarpal and metatarsal fractures are usually amenable to external coaptation using a short or full limb polyurethane resinimpregnated knitted fiberglass fabric cast. The prognosis for long-term pain-free survival is excellent for closed fractures and fair to good for open fractures managed in the manner. Surviving animals generally are not lame and do not demonstrate significant limb deformity or limb shortening and generally become completely productive. Even considering the narrow profit margin involved when treating cattle with serious injuries, this method of fracture management is usually economically and technically feasible. PMID- 8705803 TI - Orthopedic conditions of small ruminants. Llama, sheep, goat, and deer. AB - Diagnosis and treatment of diseases of the foot, infectious arthritis, angular limb deformities, patellar luxation, tendon contracture and injuries, and fractures encountered in sheep, goats, llamas, and deer are reviewed. These species share similar orthopedic problems to cattle, but management conditions, particularly for pet animals, may place special demands on the veterinarian treating these disease conditions. The mild temperament and relatively small body size of these animals make them excellent candidates for treatment of orthopedic problems often not amenable to practical treatment in larger or more fractious animals. PMID- 8705804 TI - Arthroscopy in food animal practice. AB - Arthroscopy can be an effective technique for diagnosis and treatment of various orthopedic disorders in cattle, including septic arthritis and osteochondrosis. Cost of equipment and surgery may be prohibitive for some individuals, yet the techniques of arthroscopic surgery are proven efficacious when applied to cattle. Arthroscopic techniques may also be quite useful in the evaluation and treatment of the same disorders in sheep, goats, and camillids. The technology remains the same and applicability rests with financial constraints and the physical aspects of specific species and joints. We speculate that arthroscopy would be quite useful for exploration and treatment of the larger joints and other selected joints of these species. PMID- 8705805 TI - Amputation and prosthesis. AB - Amputation of a limb is a drastic therapy in cattle and should be considered as a last resort in the treatment of limb injuries. Even though the surgical technique for amputation is simple, daily limb management, prosthesis manufacture, and adaptation of the animal to the prosthesis are difficult and time consuming. Dedication of the owner to the aftercare required for the rest of the animal's life is extremely important. Amputation of a limb with prosthesis application is a method of saving valuable animals for breeding purposes or animals of sentimental value. The author reviews his experience with partial foreleg amputation and prosthesis application in three heifers. PMID- 8705806 TI - Clinical applications of radiography and ancillary imaging. AB - Radiography will remain the most popular imaging modality in the near future for many reasons. It is economical, portable, and familiar to large animal practitioners. Contrast radiography allows evaluation of vascular and tract lesions. Other modalities may offer various advantages. Diagnostic sonography has the advantage of being able to evaluate soft tissue structure damage, fluid pockets, and lucent foreign bodies. Xeroradiography and CR use conventional, high powered x-ray units for the production of images on nontraditional ionized film plate-cassette systems. Both systems provide superior bone-soft tissue contrast, by virtue of edge enhancement, than does conventional radiography. CT and MR imaging provide cross-sectional imaging of bones and adjacent structures, avoiding structures that may be superimposed on the area of interest with conventional radiography. The major limitation is the demand for a custom engineered couch able to withstand the weight of an adult bovine. Scintigraphy provides important functional information for the clinical evaluation of orthopedic disease. It may be the most useful imaging tool to localize the cause of lameness. PMID- 8705807 TI - Surgical management of digit disorders in cattle. AB - Digital diseases in cattle are common. Cattle affected by digital disorders do not always respond to medical therapy and often need surgery. A thorough examination of the digits and radiographic evaluation help the clinician to choose the appropriate treatment and surgical approach. Age, sex, weight, type of production, and environment of the cattle also should be considered in choice of therapy. PMID- 8705808 TI - Fractures and luxations of the pelvis and proximal femur. AB - Although lameness caused by fractures and luxations of the pelvis and proximal femur are rare, they are serious and often end an animal's productive life unless properly diagnosed and promptly treated. Diagnosis of these injuries is difficult without special radiographic equipment; however, careful physical examination will often lead to a tentative diagnosis. Coxofemoral luxations and slipped capital epiphysis are two of these life-threatening injuries. This article discusses their diagnosis and possible options for treatment, which can lead to a useful, productive life. The diagnosis of other pelvic injuries is discussed, although these injuries are often not as amenable to treatment. PMID- 8705809 TI - Stifle injuries in cattle. AB - Lameness of the hindlimbs originating from the stifle joint may be difficult to diagnose and treat. This article describes anatomic considerations and the diagnostic procedures necessary to accurately evaluate the bovine stifle joint. The emphasis is toward obtaining a diagnosis based on physical examination and simple diagnostic procedures in field situation. Specific treatments and/or management alternatives for cattle with various ligament injuries (cranial cruciate rupture, collateral ligament injuries), or with luxated patella, upward fixation of the patella, septic arthritis, and subchondral bone cysts are described. Current knowledge of the prognosis after various treatments is presented. PMID- 8705810 TI - Diagnosis and management of tendon disorders in cattle. AB - Diagnosis and treatment of congenital and acquired tendon disorders in cattle are discussed. A brief discussion of tendon structure, anatomy, biomechanics, and healing is included. Congenital abnormalities presented include hyperextension deformities, flexural deformities, arthrogryposis, and spastic paresis. Acquired abnormalities discussed include tendon displacement, tendinitis, tendon disruption, and tenosynovitis. Antibiotic selection for musculoskeletal infections is briefly discussed. PMID- 8705811 TI - [Training in cognitive functions in neurologic rehabilitation of craniocerebral trauma]. AB - This study evaluates a new cognitive rehabilitation therapy for patients after severe head injury. In addition to the standard neurological rehabilitation therapy, one group was trained by the Wiener Determinationsgerat (WDT), a second group was treated by the new program REHACOM, while a third group received only conventional neurological rehabilitation therapy. The three groups each consisted of 12 patients; two groups received 20 sessions of training, each lasting 40 minutes. At the beginning as well as after the therapy a psychological test battery was applied, consisting of HAWIE, TULUC, AACHENER APHASIETEST, and BENTON Test. They were also tested by a specific neuropsychological battery regarding hemispheric specialization. REHACOM showed significantly higher values on the HAWIE as well as on BENTON-Test than the other two groups. REHACOM also improved in right-hemispheric dimensions while WDT group did not improve in attention. Right-hemispheric training was more effective than attentional stimulation. PMID- 8705812 TI - [Stimulus and reaction patterns in serial choice reactions]. AB - In serial choice-reaction-tasks reaction times decrease faster if the sequence of stimuli is structured rather than of being random. Nevertheless, the issue whether this structure-specific improvement of performance is due to the structure of the stimulus- or the response-sequences is controversially discussed. After a review of the corresponding literature, an experiment will be reported which was designed to contribute to this issue. Contrary to the present experiments, which exclusively analysed effects of statistical redundancy, relational patterns are introduced into the sequences of stimuli as well as into the sequences of responses. These relational patterns were varied independently of each other. The data reveal strong effects of the relational patterns in the sequence of responses, whereas the relational patterns of the stimulus sequences were not systematically effective. Furthermore, the data suggest that the relational structure of the response sequences especially favours the formation of "motor-chunks". In the discussion it is emphasized that effects of relational patterns on serial choice reactions are not yet adequately regarded, neither in experimental research nor in the theoretical accounts. PMID- 8705813 TI - [Modification of performance by activation in the elderly]. AB - The effects of activation on performance are rarely investigated in older people. An experiment on the effects of psychic activation on free recall of word lists is reported. Participants were healthy women (N = 71) of two age groups (60-69 versus 70-79 years). Psychic activation was induced by intermittent white noise of three intensities (55, 80, 90 dBA). The effects were assessed by self-report inventories concerning mood and performance, by vegetative variables, and by memory tests. The memory tests consisted of free recall tasks under instructions of power and speed. Under the power condition memory performance decreased with activation in both age groups. Free recall performance under speed conditions resulted in inverted U-shaped relations between activation and performance only for the younger sample; the older group was not affected. To explain the results possible mediators are discussed. PMID- 8705814 TI - Extending the scope of gingival crevicular fluid elastase research. PMID- 8705815 TI - The relationship between elastase and lactoferrin in healthy, gingivitis and periodontitis sites. AB - OBJECTIVES: To compare the relative amounts of elastase (primary polymorphonuclear leucocyte granule constituent) and lactoferrin (secondary PMN granule constituent) in the gingival crevicular fluid (GCF) of healthy, gingivitis and periodontitis sites. DESIGN: This cross-sectional study looked at the two GCF constituents in three categories of disease status within the same subject. MATERIALS AND METHODS: Patients with chronic adult periodontitis were screened and those exhibiting all three types of sites ie periodontally healthy, gingivitis and periodontitis sites were recruited (n=10) and had GCF collected from the three sites. Lactoferrin and elastase were measured in eluates of GCF by enzyme-linked immunosorbent assay. RESULTS: The absolute amount of lactoferrin measured in ng per 30 s samples was significantly lower in healthy and gingivitis sites as compared to periodontitis sites; however this difference failed to reach significance when the concentration of lactoferrin in GCF was used as the analytical unit. No significant differences were found for elastase levels at any sites when expressed as either absolute amounts or concentrations. Secondary granule release, as evidenced by lactoferrin levels, occurs during cell migration and the process is independent of primary granule release, which is thought to correlate with PMN activation. The relationship between granule constituents in the samples showed significant differences, the highest lactoferrin/elastase ratio being at periodontitis sites (P<0.001). CONCLUSIONS: These findings imply a change in the relative amounts of elastase and lactoferrin released at different disease level sites, wth an almost 10-fold increase in the proportion of lactoferrin to elastase in periodontitis sites over healthy and gingivitis sites. This variation in the release by PMNs of primary and secondary granule constituents may indicate alterations in PMN function in different disease environments. PMID- 8705816 TI - Elastase in gingival crevicular fluid from smokers and non-smokers with chronic inflammatory periodontal disease. AB - OBJECTIVE: To compare elastase concentrations in gingival crevicular fluid (GCF) from individual sites of smokers and non-smokers. MATERIALS AND METHODS: Twelve pairs of smokers and non-smokers with untreated, moderate to advanced chronic inflammatory periodontal disease were matched for gender, age, ethnicity and the clinical and radiographic extent of disease. Durapore filter strip samples were collected over 30 s from two mesiopalatal sites on upper left posterior teeth. Samples were analysed for: 1) polymorphonuclear neutrophil leucocyte (PMNL) cell counts; 2) PMNL elastase-alpha 1-antitrypsin complex in the GCF supernatant by ELISA; and 3) functional elastase, free or bound to alpha 2-macroglobulin, estimated from activity against N-tert-butoxycarbonyl-alanyl-prolyl-norvalyl-p chlorothiobenzyl ester in supernatant and lysates of GCF PMNLs. RESULTS: There were no differences in disease parameters between groups except that bleeding on probing was less extensive in smokers (P<0.001). Cell counts and elastase content of crevicular PMNLs showed no differences between groups. Lower concentrations of elastase were found in GCF supernatants from smokers than non-smokers. This difference was observed for functional elastase (mean [s.d.] = 30.21 [17.60] against 73.77 [75.26] ng microliter(-1), p<0.05) and elastase complexed with alpha 1-antitrypsin (8.97 [6.54] ng microliter(-1) against 25.71 [22.07] ng microliter(-1), p<0.001). CONCLUSIONS: Smokers have lower elastase concentrations in GCF than non-smokers. Further investigation is required to elucidate the underlying cause and its relationship with periodontal disease. PMID- 8705817 TI - Comparative virulence of periodontopathogens in a mouse abscess model. AB - OBJECTIVE(S): This report compares the virulence of selected strains of P. gingivalis, A. actinomycetemcomitans, C. rectus, F. nucleatum and T. denticola in a murine model as a measure of pathogenic potential of these oral microorganisms. The characteristics of the tissue destruction associated with these monoinfections were then related to a potential model for bacterial synergism in progressing periodontitis. DESIGN AND METHODS: All bacterial strains were grown to mid-logarithmic to early stationary growth phase, harvested and used at various doses to challenge BALB/c normal and BALB/c dexamethasone (DEX) treated mice to mimic a neutrophil dysfunction. The characteristics of tissue destruction, and overt tissue destructive capacity of these species were examined as a function of challenge dose and time. OUTCOME MEASURES: The mice were examined for an interval of approximately 15 days post-challenge and the presence/absence of lesions, localized or generalized nature of the lesion (including size in mm2), and lethality of the infection were assessed. RESULTS: Comparison of the virulence of the various P. gingivalis strains related to lethality and lesion size associated with destruction of the connective tissue, indicated a virulence capacity of P. gingivalis strains 53977>W50 = T22>3079>33277>381. C. rectus elicited localized necrotic lesions which were limited to the epithelial layers of the skin. The size of the lesions also indicated a graded difference in virulence, such that C. rectus strains 234>576>>33238. A. actinomycetemcomitans caused the formation of classic localized abscesses with a PMN infiltrate and inflammatory exudates. Although each of the A. actinomycetemcomitans strains exhibited a similar virulence pattern in this murine model, A. actinomycetemcomitans serotype b representative strains were potentially more pathogenic with a virulence capacity of 3113D-N = 3975A>JP2 > or = Y4>29523>33384. Both C. rectus and A. actinomycetemcomitans strains showed clear evidence that recent clinical isolates were more virulent than laboratory strains. Challenge with F. nucleatum resulted in tissue destructive responses which were different from those observed with the other strains used in this study. A rapid onset of dose-dependent lesion development, related to the formation of either closed abscesses or open lesions, was observed with F. nucleatum. Tissue involvement was also greater at lower F. nucleatum doses when compared to the other bacteria. F. nucleatum challenge of DEX-treated mice resulted in a shift to open lesions. T. denticola appeared to be more tissue invasive than the other species examined in this study. Challenge of mice with T. denticola resulted in involvement of multiple tissues, including epithelial and connective tissues, as well as appearing to invade muscle layers and deeper tissues. In addition to invading deeper tissues, the resulting lesions took considerably longer to resolve. In the DEX-treated mice (neutrophil depleted), P. gingivalis, C. rectus, and A. actinomycetemcomitans were significantly more virulent. In contrast, while DEX treatment altered the characteristics of lesions caused by F. nucleatum, the extent of lesions produced by F. nucleatum and T. denticola was not substantially enhanced. CONCLUSIONS: The results obtained from this study suggest that different microorganisms have the ability to provide individual pathologies which may act in an additive/synergistic fashion contributing to the tissue destruction noted in periodontitis. PMID- 8705818 TI - Longitudinal dynamics of infection and serum antibody in A. actinomycetemcomitans periodontitis. AB - OBJECTS: This report describes one of the first prospective studies delineating the relationship between infection, host antibody responses and disease exacerbations and remissions in a distinct subset of periodontitis patients infected with A. actinomycetemcomitans. DESIGN: The design of this longitudinal study included visits for each patient approximately every 2 months for up to 3 years. SUBJECTS AND METHODS: Subjects (n=51) included 16 adult periodontitis (AP) and 11 early-onset periodontitis (EOP) patients with elevated serum IgG antibody to A. actinomycetemcomitans and infection with this microorganism, 12 AP patients with normal levels of anti-Aa antibody, and 12 normal subjects. MEASUREMENT OUTCOMES: Clinical parameters included a gingival index, plaque index, bleeding on probing, pocket depth, and attachment level. Disease activity was defined as loss of attachment during the monitoring intervals. Serum IgG, IgM and IgA antibody to A. actinomycetemcomitans Y4 (serotype b) was quantitated using an ELISA. Subgingival plaque samples were examined for A. actinomycetemcomitans using colony immunoblotting. Human serum IgG antibody specificities to outer membrane antigens (OMA) of A. actinomycetemcomitans Y4 were determined using Western immunoblotting. RESULTS: A. actinomycetemcomitans-infected AP patients had a higher frequency of teeth infected when compared to the EOP patients. However, the EOP patients exhibited a trend for higher levels of A. actinomycetemcomitans in those teeth that were infected. Active disease patients demonstrated a significantly greater frequency of infected sites, as well as significant elevations in the proportions of A. actinomycetemcomitans. Both EOP and AP groups showed significantly elevated IgG, IgM and IgA antibody to A. actinomycetemcomitans when compared to a periodontally normal group. The level of IgG antibody was significantly elevated in A. actinomycetemcomitans-positive patients with active disease, while IgA antibody was decreased in a number of the active group patients. Plaque samples derived from active sites showed a clear and significant increase in A. actinomycetemcomitans that occurred from 2-6 months prior to the identification of disease activity. Approximately 70% of the active disease patients showed an increase in IgG antibody level by 2-4 months prior to disease activity. Studies of the antigen reactivity patterns of serum IgG indicated that antibody to antigens of 65, 58, 48, 29 and 24 kDa were more frequent in patients who showed active disease, while those patients with the greatest frequency of active disease appeared to show a general decrease in the recognition of the A. actinomycetemcomitans OMA. CONCLUSIONS: It appears that A. actinomycetemcomitans infection relates to a particular type of disease with accompanying antibody responses that reflect periods of active disease. The dynamics of A. actinomycetemcomitans infection and the level and specificity of systemic antibody responses to this pathogen support an important contribution of the immune response to managing this infection. PMID- 8705819 TI - Cytokine profiles of Porphyromonas gingivalis-reactive T lymphocyte line and clones derived from P. gingivalis-infected subjects. AB - Porphyromonas gingivalis is generally recognized as a major periodontopathogen such that a study of T cell responses to this organism may help to elucidate immune regulation in periodontal disease. OBJECTIVE: The aim of this study was to examine interleukin (IL)-4, interferon (IFN)-gamma and IL-10 production by P. gingivalis-responsive T cell lines and clones derived from the peripheral blood of two P. gingivalis-infected subjects with different disease expression and from the gingival tissues of one of the P. gingivalis-infected subjects. MATERIALS AND METHODS: FACS analysis was used to determine the percentage of T cells staining positive for cytoplasmic IL-4, IFN-gamma and IL-10 and reverse transcriptase polymerase chain reaction (RT-PCR) was performed to determine the presence of mRNA for IL-4 and IFN-gamma in the T cell lines and clones. RESULTS: FACS analysis showed that virtually all the T cell lines and clones contained IL-4- and IFN-gamma-producing T cells. The RT-PCR results generally supported this trend. However, a higher percentage of cells in the clones derived from one subject produced IL-4 while a lower percentage produced IFN-gamma compared with the clones derived from the other subject. FACS analysis also demonstrated that the lines and clones derived from the two subjects showed differences in IL-10 production. CONCLUSION: This study has demonstrated that there may be differences in IL-4 and IL-10 production by the P. gingivalis responsive lines and clones derived from P. gingivalis-infected subjects with different disease expression. Any relationship to disease however, remains to be determined. PMID- 8705820 TI - Oral disease in the elderly in long-term hospital care. AB - OBJECTIVE: To obtain baseline information on the prevalence of oral problems and disease in institutionalized elderly in a Scottish hospital. DESIGN: A cross sectional clinical investigation with complementary microbiological studies as appropriate. SUBJECTS AND METHODS: A cohort of 147 elderly in five long-term care wards; collection of demographic data; clinical examination to determine: i) the dental and denture status and associated lesions such as Candida-associated denture stomatitis and angular cheilitis, ii) oral mucosal disease; swabs as appropriate for microbiology. MAIN OUTCOME MEASURES: Dental status, root caries prevalence; denture status and hygiene, and associated disease; oral mucosal health. RESULTS: Median period of institutionalization of 147 patients was 15 months and 65% were aged 80 years or more; the majority were significantly mentally impaired; 52% wore complete dentures, 9% were partially dentate, 19% possessed partial or incomplete dentures; 20% were neither dentate nor had dentures. Of 80 patients who verbalised their complaints, 35% complained of dry mouth and 45% had denture-related problems, principally discomfort. The commonest oral finding was a coated tongue (56%); angular cheilitis was present in 25%, and Candida-associated denture stomatitis in 19%; none had oral ulcers. CONCLUSIONS: There was considerable unmet dental need with significant oral disease and poor levels of oral and denture hygiene in this target group; both dental and medical professionals should act in concert to deliver curative as well as preventative dental care for the elderly living in institutions to improve their quality of life. PMID- 8705821 TI - The histopathology of oral mucosal lesions associated with amalgam or porcelain fused-to-metal restorations. AB - OBJECTIVES: To analyse the interface stomatitis patterns of oral lichenoid lesions in contact with amalgam and to compare these with the histologic changes in oral lesions clinically associated with porcelain-fused-to-metal (PFM) restorations. To relate these features to the presence of tissue-bound mercury (Hg). DESIGN: A retrospective analysis of tissue biopsies, with clinical data collected via a complementary questionnaire. SUBJECTS AND METHODS: 479 biopsies diagnosed in 1987 as 'lichenoid reactions'. From these, we retrieved all with amalgam contact and without candida or medication. From 1990-91, all mucosal lesions stated to be associated with PFM restorations were then retrieved for comparative analysis. The biopsies were examined with routine histologic and autometallographic methods. RESULTS: 77 amalgam-associated lesions were found and could be subdivided into five pre-defined interface stomatitis types. We found 22 lesions associated with PFM and 20 showed histopathologic features similar to those associated with amalgam. Hg accumulations were detected in the majority of amalgam-associated but only in part of the PFM-associated lesions. CONCLUSIONS: Amalgam-associated lichenoid lesions present a wide spectrum of histopathologic patterns, corresponding to similar patterns in dermatopathology but with no evidence of association with specific disease. PFM-associated lesions tend to display similar lichenoid features, suggestive of common pathogenetic mechanisms. Hg accumulations may play a role to maintain the chronicity of such lichenoid lesions. PMID- 8705823 TI - Two cases of tongue cancer treated with intra- and peri-tumoral injection of recombinant interleukin-2 alone: immunohistochemistochemical considerations to clinical tumor response. AB - SUBJECTS AND METHODS: Two cases with stage II tongue cancer who exhibited different responses to intra- and peri-tumoral administration of rIL-2 alone are presented. Special consideration is given to the relationship between tumor responses to rIL-2 and clinicopathological and immunohistopathological findings. RESULTS: The patient who responded completely to treatment showed an exophytic tumor growth pattern, low-grade cancer invasion, and predominant infiltration of CD8+ lymphocytes over CD4+ lymphocytes in cancer cell nests. The non-responder showed endophytic tumor growth, high-grade cancer invasion, and uniform distribution of both CD4+ and CD8+ lymphocytes in cancer cell nests. CONCLUSIONS: Distribution of adequate amounts of T lymphocytes subsets may be necessary in order for good tumor response to biotherapy with rIL-2; other clinical and histopathological variables predicting the effect for cancer chemotherapy remain to be identified. PMID- 8705822 TI - Changes in the expression of integrins and basement membrane proteins in benign mucous membrane pemphigoid. AB - OBJECTIVE: To determine the location of the subepithelial split in benign mucous membrane pemphigoid (BMMP) and its relationship to the anchoring filaments and their receptors. MATERIALS AND METHODS: Frozen sections of lesional and perilesional oral mucosa from 10 cases of BMMP were stained, using an immunofluorescence method, for the beta-1, beta-4, alpha-3 and alpha-6 integrin subunits and for their ligands, laminin I and laminin V (kalinin). In all cases the diagnosis was confirmed by the demonstration of linear staining for IgG at the basement membrane zone. Six specimens of normal mucosa were stained for comparison. RESULTS: Staining for integrins, laminin and kalinin in perilesional mucosa was similar to normals, although one case showed loss of alpha-6 and beta 4 . In lesional mucosa, laminin and kalinin showed strong linear staining localised to the floor of the bullae. The alpha-6 and beta-4 subunits were expressed only on the roof of the bullae but staining was weak and patchy with areas of loss. In some sections alpha-6 showed a punctate intracellular distribution similar to IgG. The distribution of alpha-3 and beta-1 was similar to that seen in normals. CONCLUSIONS: In all cases kalinin was found on the connective tissue side of the lesions and alpha-6 beta-4 localised to the epithelial side. This shows that the split occurs at a location which separates anchoring filaments from the hemidesmosomes. Loss of the alpha-6 beta-4 integrin in the lesions and the similar intracellular staining of alpha-6 and IgG, suggest that disruption of hemidesmosomes may be a key event in the immunopathogenesis of the lesions and that the alpha-6 integrin subunit is a potential antigen in oral mucosal BMMP. PMID- 8705824 TI - Review article: The role of oncogenes, tumour suppressor genes and growth factors in oral squamous cell carcinoma: a case of apoptosis versus proliferation. AB - Mutation, deactivation and disregulated expression of oncogenes and tumour suppressor genes may be involved in the pathogenesis of oral squamous cell carcinoma (SCC). Deactivation of the p53 tumour-suppressor gene allows cell proliferation and blocks apoptosis of malignant oral keratinocytes. Mutation in the ras oncogene results in persistent mitogenic signalling. Upregulatioed c-Myc expression, in the presence of growth factors, provides an additional proliferative signal. Loss of retinoblastoma tumour-suppressor gene (Rb) function may contribute to oral keratinocyte hyperproliferation and recent evidence suggests that simultaneous deactivation of both p53 and Rb is required for tumourigenesis. Enhanced Bcl-2 and reduced Fas expression inhibit tumour cell apoptosis and may convey resistance to cytotoxic drugs and T cell-mediated cytotoxicity, respectively. Exogenous mutagens such as tobacco, alcohol and viral oncogenes may cause altered expression of oncogenes and tumour-suppressor genes in some cases of oral SCC. The impact of these mechanisms on future therapies for oral SCC is highlighted. PMID- 8705825 TI - Cell biology of the periodontium in health and disease. PMID- 8705826 TI - Expression of lymphocyte function-associated antigen 3 in oral lichen planus. AB - OBJECTIVE: The expression pattern of lymphocyte function-associated antigen 3 (LFA-3) in the buccal mucosa of oral lichen planus (OLP) patients was compared to that of healthy controls to investigate the possible role of LFA-3 in cell interactions within OLP lesions. MATERIALS AND METHODS: Samples of buccal mucosa from 17 clinically healthy individuals and 17 OLP lesions were analysed. Expression of LFA-3, CD2, CD3 and CD14 was visualized by an immunoperoxidase technique and assessed microscopically. RESULTS: In healthy buccal mucosa LFA-3 was expressed on keratinocytes, Langerhans cells within the epithelium and on endothelial cells in the lamina propria. In OLP patients a similar pattern of LFA 3 staining was observed. In addition, cytoplasmic LFA-3 without accompanying surface staining, was seen on a subpopulation of macrophage-like cells. Substantial amounts of LFA-3 also appeared to be associated with non-cellular components of the extracellular matrix within the inflammatory infiltrate. CONCLUSIONS: We have obtained evidence for a previously undescribed localization of LFA-3 within macrophages, and have observed that expression of LFA-3 is apparently elevated within OLP lesions. LFA-3 may play an important role in the pathogenesis of OLP. PMID- 8705827 TI - Cross-infection compliance of UK dental staff and students. AB - OBJECTIVES: The objectives of the study were to determine the compliance of a group of dental health care workers and students in aspects of cross-infection control. DESIGN: The dentists and dental students working in a dental clinic were observed by a 'hidden' ceiling-mounted video camera. Procedures were recorded onto videotape and the actions of the observed clinicians and students assessed by two suitably qualified assessors. SETTING: An emergency Oral Medicine clinic in a dental hospital. SUBJECTS: Seventy nine dentists and 35 dental undergraduates. METHODS: The action of staff and students were recorded on videotape and assessed retrospectively. MAIN OUTCOME MEASURES: Degree of compliance with recognised infection control policies. RESULTS: Compliance with recommended guidelines for control of cross-infection was poor. In only 56% of health care worker-patient contacts were protective gloves changed between patients. Facemasks and protective eyewear were only worn in 38% and 29% of such contacts respectively. CONCLUSIONS: Compliance with cross-infection control measures are poor in dentistry--even when clinicians are provided with appropriate facilities. PMID- 8705828 TI - Correlation between gingival crevicular fluid dipeptidyl peptidase II and IV activity and periodontal attachment loss. A 2-year longitudinal study in chronic periodontitis patients. AB - OBJECTIVE: The aim of this study is to determine whether gingival crevicular fluid (GCF) dipeptidyl peptidase (DPP) II or IV levels, total activity (TA) and concentration (EC), predict progressive attachment loss (AL). SUBJECTS AND METHODS: Seventy five patients with moderate periodontitis were recruited. GCF was first collected from 16 molar and premolar mesiobuccal sites and then probing attachment level (PAL) and probing pocket depth (PPD) were measured with an electronic probe. Finally, gingival index, gingival bleeding and plaque indices were scored. Patients were given basic periodontal treatment prior to baseline after which the above procedures were repeated. Patients were seen 3 monthly for 2 years and the procedures were repeated. Carefully localised radiographs were taken of the test teeth annually. RESULTS: One hundred and twenty AL sites, 88 rapid AL (RAL) and 32 gradual AL (GrAL), in 48 patients were detected. DPP II and IV levels (TA and EC) at RAL sites were significantly higher (P < or = 0.0001) than paired control sites at the attachment loss time (ALT) and prediction time (PT). Mean levels over the study period of both proteases (TA and EC) at GrAL sites were significantly higher (P < or = 0.0001) than other paired control sites. The GCF levels of DPP IV were always slightly higher than those of DPP II. Critical values (CV) of 5 microU per 30 s (TA) and 25 microU microL(-1) (EC) for both proteases showed high sensitivity and specificity values for TA and EC and these were the same at both ALT and PT. The positive predictive values were slightly higher for DPP II. Mean site DPP II and IV levels (TA and EC) in intra patient comparisons were significantly higher (P < or = 0.0001) at RAL and GrAL sites than non-attachment loss (NAL) sites in AL patients and mean patient levels were significantly higher (P < or = 0.0001) in AL patients than NAL patients in inter-patient comparisons. CONCLUSIONS: These results indicate that both GCF DPP II and IV may be predictors of periodontal attachment loss. PMID- 8705829 TI - Diabetic periodontitis: possible lipid-induced defect in tissue repair through alteration of macrophage phenotype and function. AB - BACKGROUND: Diabetes mellitus is a major health problem in the United States affecting approximately 13 million people. The five 'classic' complications which have historically been associated with the condition are microangiopathy, neuropathy, nephropathy, microvascular disease, and delayed wound healing. Recently, periodontal disease (PD) has been declared the 'sixth' major complication of diabetes as diabetics demonstrate an increased incidence and severity of PD. The cellular and molecular basis for diabetic PD is unknown. HYPOTHESIS: Recent evidence suggests that PD and delayed dermal wound healing may be manifestations of the same general systemic deficit in diabetes involving impairment of the cellular and molecular signal of wounding via alterations in macrophage phenotype. Diabetes-induced hyperlipidemia may interfere with the normal cellular and molecular signal of wounding by alteration of macrophage function and subsequent dysregulation of cytokines at the wound site. RESULTS: Preliminary data in both animal models and humans suggests that hyperglycemia, in combination with elevations of serum low density lipoproteins and triglycerides, leads to formation of advanced glycation end products (AGEs) which may alter macrophage phenotype. This may be responsible for dysregulation of macrophage cytokine production and increased inflammatory tissue destruction and alveolar bone loss. IMPLICATIONS: Future investigations will consider diabetic PD in the context of a generalized systemic wound healing deficit that manifests as PD in the face of constant pathologic wounding of the gingiva (bacterial plaque) or delayed dermal wound healing in instances of periodic traumatic wounding to other parts of the body. These types of studies will provide information concerning defective tissue repair in diabetics that will have clinical relevance for the understanding of PD and delayed dermal healing as well as applications of appropriate and specific therapies. PMID- 8705831 TI - Cell Biology of the Periodontium in Health and Disease. Proceedings of the Symposium at University College, Cork, September 1995. PMID- 8705830 TI - The soft connective tissues of the gingiva and periodontal ligament: are they unique? AB - The connective tissues of the gingiva and periodontal ligament share a common embryonic development from cells of the cranial neural crest. This review paper describes the relationship of these tissues in tooth germ initiation, development and eruption. PMID- 8705832 TI - Turnover in periodontal connective tissues: dynamic homeostasis of cells, collagen and ground substances. AB - The connective tissues of the periodontium are composed of two soft tissues and two hard tissues--each of which has unique features. This review considers the constituents of normal, healthy periodontal connective tissues together with an appraisal of the changes in the connective tissue matrices of the periodontium which occur during the development of periodontitis. Recent developments in this field have paved the way for new and exciting vistas in periodontal diagnosis and regeneration which, ultimately, are two important goals in periodontal therapy. PMID- 8705833 TI - Periodontal inflammation and attachment loss: a critical problem for biological studies. AB - BACKGROUND: In biological studies of periodontitis, there has been long-standing confusion between the ubiquitous phenomena of inflammation and the essential criterion of attachment loss. This is partly attributable to inadequate methods of clinical measurement, but seems also to be a consequence of an unproven and usually unstated assumption that the same biological processes underlie both inflammation and attachment loss. Developments in unidimensional clinical probing methods have helped in studies of periodontal treatment. However, such methods are intrinsically unsuitable in studies of periodontal diseases, and may have given them a false sense of security. the confusion has been compounded by inappropriate use of statistical techniques in an attempt to solve problems which do not arise from mathematical models but are intrinsic to measurement methods. OBJECTIVE: This paper is a clinician's attempt to state the current difficulties and suggest some ways forward, including the development of three-dimensional measurement, non-invasive probing, and several objectives for biochemical, microbiological and immunological research. PMID- 8705834 TI - Cytokines and cell signalling in the periodontium. AB - A large number of potential regulatory mechanisms have been described which may be involved in the control of cell function in the periodontium. In this review, soluble effector molecules which may regulate normal cell turnover and which may control the maintenance of the periodontal space are considered. There is evidence for the involvement of growth factors including EGF, PDGF, FGFs, IGF I & II and TGF-beta in these processes. The role of bone morphogenetic proteins (BMPs) in periodontal turnover is of considerable interest as they appear to be able to regulate all stages of this process from specifying cell commitment to regulating differentiated cell function. Empirical evidence suggests the importance of mechanical stimulation in controlling the maintenance of the periodontal ligament space. The wide range of effects of mechanical stimulation are briefly reviewed and the central role of prostaglandins is considered. Recent evidence suggests the involvement of nitric oxide (NO) in the regulation of mineralised tissue function, and the potential role of NO in maintenance of the ligament space is considered. Further studies are required which address the interactions between all of these mechanisms in order to determine the key factors which may control periodontal cell function. For the future an understanding of these interactions has the potential to lead to important clinical developments in periodontal and orthodontic therapy. PMID- 8705835 TI - Cytokine networks in destructive periodontal disease. AB - BACKGROUND: Cytokines are important regulatory proteins, produced by activated cells, which act by binding high affinity cell surface receptors. They are involved in almost all aspects of cell biology and form interacting networks, with cascades of sequential cell activation. They often show overlapping activities (redundancy) or the same cytokine may have a variety of different effects (pleiotropy). In excess, certain cytokines are damaging and proinflammatory. Tumour necrosis factor alpha (TNF alpha) and interleukin-I (IL I) are markedly proinflammatory, inducing bone resorption, collagenase and prostaglandin E2 production. OBJECTIVE: This paper focuses on the role of TNF alpha and IL-I in the cytokine networks of destructive chronic periodontitis; specifically their regulation by T cell cytokines, receptor antagonists and inhibitory soluble forms of the IL-I and TNF receptors. CONCLUSION: A hypothesis is proposed that destructive periodontal disease may be due to disregulation of these inhibitors, rather than an overproduction of IL-I and TNF alpha per se. PMID- 8705836 TI - Origins and functions of cells essential for periodontal repair: the role of fibroblasts in tissue homeostasis. AB - OBJECTIVE: A review is undertaken of rodent model systems and cell culture studies that address the role of periodontal fibroblasts in tissue homeostasis in both normal function and after wound healing. RATIONALE: Fibroblasts are the predominant cells of the periodontal ligament (PL) and of healthy gingiva and have important roles in the development, function and regeneration of the tooth support apparatus. REVIEW: In normally functioning periodontal tissues cell turnover involves generation of new cells by proliferation which in turn is balanced by apoptopic cell death. Consequently PL fibroblasts comprise a renewal cell system in steady-state. PL cell progenitors can generate multiple types of more differentiated, specialized cells including large numbers of fibroblastic cells and more limited numbers of osteogenic or cementogenic cells. However PL fibroblasts constitutively block osteogenesis and thereby maintain the PL width. Proliferating progenitor cell populations of the PL are enriched in locations adjacent to blood vessels and in contiguous endosteal spaces from where they migrate to the body of the PL. Large increases of cell formation and cell differentiation occur after wounding but surprisingly, the cells that repopulate the PL adjacent to the root surface are largely post-mitotic. As PL cell populations comprise multiple lineages, it is likely that after wounding, the separate phenotypes repopulating the wound site will be selected by environmental factors. Further, the specific repopulating lineages will strongly influence the form and function of the nascent tissue. To illustrate the specificity of fibroblast functions, examples of migratory and contractile fibroblast phenotypes are provided which exhibit constitutively different levels of gelsolin and alpha smooth muscle actin respectively, cytoskeletal proteins which are markers for these cell types. CONCLUSION: Fibroblasts contribute to PL homeostasis by their abilities to remodel tissues, to repopulate wounds, to influence the metabolism of other cell types and to create a new fibrous attachment. PMID- 8705837 TI - Modulation of host matrix metalloproteinases by bacterial virulence factors relevant in human periodontal diseases. AB - OBJECTIVE: Bacterial pathogens involved in periodontal diseases exert a part of their destructive effect by triggering and inducing host cells to elevate their secretion of matrix metalloproteinases (MMPs). Pathogen-secreted phospholipase (PLC) is one bacterial product that may trigger this host response. The roles of exogenous PLC leading to the release, secretion and expression of MMPs by peripheral blood neutrophils (PMNs), cultured epithelial cells of human gingiva and porcine periodontal ligament were investigated. Also the activities of PLC in the diseased and healthy gingival sulcular fluid (crevicular fluid, GCF) and molecular forms of gelatinases present in dental plaque were investigated. MATERIALS AND METHODS: GCF, salivary and dental plaque samples were analyzed for PLC and proteinase activities. The abilities of PLC to induce PMNs and oral epithelial cells to release and express their MMPs were examined by specific functional, immunological and molecular biology means. RESULTS: PMN-derived MMPs were found to predominate in periodontitis GCF and plaque, and PLC activities were higher in GCF of adult periodontitis patients than in healthy controls. Purified bacterial PLC (1 mU ml(-1)) efficiently induced PMN degranulation. PLC also induced MMP expression in the cultured epithelial cells. The strongest response was seen in MMP-9 and less in MMP-2. The induction was dose-dependent in the range of 0.0-1.0 U ml(-1) PI-PLC, and quiescent cultures were more responsive than proliferating ones. PLC induction of MMPs was polar, with increased levels of MMP-9 in the apical region and increased MMP-2 levels secreted in the basal direction. Northern analysis showed a strong increase in mRNA levels of MMP-9 and a smaller increase for MMP-2 and MMP-1. In the second part of the study we investigated the molecular forms of the released MMPs during periodontitis. In bacterial plaque of periodontitis patients the MMP-9 were found to be converted into lower molecular weight forms. Isolated proteinase from Porphyromonas gingivalis (ATCC 33277) was able to convert human proMMPs to their active forms. CONCLUSION: Bacterial PLC may induce degranulation of PMN MMPs and increase MMP expression in oral epithelial cells. The released proteases can be converted into active form by the proteases of plaque bacteria. Thereby, the pathogenic oral bacteria may indirectly participate in the destruction of periodontal tissues. PMID- 8705838 TI - Vindication is sweet, but not sufficient. PMID- 8705839 TI - Plasma HIV viremia and viral load in lymph nodes. PMID- 8705840 TI - A closer look at malaria in India. PMID- 8705841 TI - The greatest threat of all. PMID- 8705842 TI - Vitamin D receptors and hyperparathyroidism. PMID- 8705843 TI - The Baltimore affair: let's not forget what went wrong. PMID- 8705844 TI - Work on your theories! PMID- 8705845 TI - Thalassemia--a global public health problem. PMID- 8705846 TI - New clues to Alzheimer's disease: unraveling the roles of amyloid and tau. PMID- 8705847 TI - Seeing how we hear. PMID- 8705848 TI - An ace up the sleeve of the cholera bacterium. PMID- 8705849 TI - Chlamydia in coronary plaques--hidden culprit or harmless hobo? PMID- 8705850 TI - Genetic vaccination: the advantages of going naked. PMID- 8705851 TI - Ventricular remodeling: hype or hope? PMID- 8705852 TI - Will there be noise in their ears? PMID- 8705853 TI - Seeking the cause of Kaposi's sarcoma. PMID- 8705854 TI - Secreted amyloid beta-protein similar to that in the senile plaques of Alzheimer's disease is increased in vivo by the presenilin 1 and 2 and APP mutations linked to familial Alzheimer's disease. AB - To determine whether the presenilin 1 (PS1), presenilin 2 (PS2) and amyloid beta protein precursor (APP) mutations linked to familial Alzheimer's disease (FAD) increase the extracellular concentration of amyloid beta-protein (A beta) ending at A beta 42(43) in vivo, we performed a blinded comparison of plasma A beta levels in carriers of these mutations and controls. A beta 1-42(43) was elevated in plasma from subjects with FAD-linked PS1 (P < 0.0001), PS2N1411 (P = 0.009), APPK670N,M671L (P < 0.0001), and APPV7171 (one subject) mutations. A beta ending at A beta 42(43) was also significantly elevated in fibroblast media from subjects with PS1 (P < 0.0001) or PS2 (P = 0.03) mutations. These findings indicate that the FAD-linked mutations may all cause Alzhelmer's disease by increasing the extracellular concentration of A beta 42(43), thereby fostering cerebral deposition of this highly amyloidogenic peptide. PMID- 8705855 TI - Glycosylation of microtubule-associated protein tau: an abnormal posttranslational modification in Alzheimer's disease. AB - Alzheimer's disease (AD) is characterized by the presence of numerous neurons with neurofibrillary tangles of paired helical filaments (PHFs). The microtubule associated protein tau in abnormally hyperphosphorylated form is the major protein subunit of the PHF. We now show that PHF tangles isolated from AD brains are glycosylated, whereas no glycan is detected in normal tau. Deglycosylation of PHF tangles by endoglycosidase F/N-glycosidase F converts them into bundles of straight filaments 2.5 +/- 0.5 nm in diameter, similar to those generated by the interaction of normal tau and abnormally hyperphosphorylated tau (AD P-tau). Deglycosylation plus dephosphorylation, but not deglycosylation alone, of AD P tau and tau from PHF tangles restores their microtubule polymerization activity. Dephosphorylation of deglycosylated PHF tangles results in increased tau release. Thus, although the abnormal phosphorylation might promote aggregation of tau and inhibition of the assembly of microtubules, glycosylation appears to be responsible for the maintenance of the PHF structure. PMID- 8705856 TI - Colocalization of retrovirus and target cells on specific fibronectin fragments increases genetic transduction of mammalian cells. AB - Hematopoietic cells are important targets for genetic modification with retroviral vectors. Attempts at human gene therapy of stem cells have achieved limited success partly because of low gene transfer efficiency. Chymotryptic fragments of the extracellular matrix molecule fibronectin used during infection have been shown to increase transduction of human hematopoietic progenitor cells. Here, we demonstrate that this enhanced gene transfer into mammalian target cells is due to direct binding of retroviral particles to sequences within the fibronectin molecule. Transduction of mammalian cells, including murine long-term repopulating hematopoietic cells, is greatly enhanced when cells are adherent to chimeric fragments containing these retroviral binding sequences. In addition, colocalization of retrovirus and target cells on fibronectin peptides allows targeted transduction of specific cell types by exploiting unique ligand/receptor interactions. PMID- 8705857 TI - Microencapsulated genetically engineered live E. coli DH5 cells administered orally to maintain normal plasma urea level in uremic rats. AB - Safety concerns about introducing genetically engineered cells into the body have prevented their use in medical treatments. To solve this problem, we prepared polymeric membrane artificial cells (semipermeable microcapsules) containing genetically engineered live cells from the bacteria Escherichia coli DH5. When given orally, the cells remain at all times in the microcapsules and are finally excreted in the stool. During their passage through the intestine, small molecules like urea diffuse rapidly into the microcapsules and are acted on by the genetically engineered cells. This lowers the high plasma urea level to normal in uremic rats with induced kidney failure, and has exciting implications for the use of this and many other types of genetically engineered cells in a number of medical applications. PMID- 8705858 TI - Vaccination against tuberculosis by DNA injection. AB - There are 3 million deaths per annum worldwide due to tuberculosis, and AIDS is compounding the problem. A better vaccine than the live mycobacterium currently in use, bacillus Calmette-Guerin (BCG), is needed. When mice were injected with plasmid DNA encoding a single mycobacterial antigen (65-kDa heat shock protein, hsp65) they made specific cellular and humoral responses to the protein and became immune to subsequent challenge with Mycobacterium tuberculosis. Protection was equivalent to that obtained by vaccinating with live BCG, whereas immunizing with the protein was ineffective. Protection was also obtained with DNA encoding another mycobacterial antigen (36-kDa proline-rich antigen). These results suggest that DNA vaccination might yield improved vaccines to replace BCG. PMID- 8705859 TI - Immunogenicity and protective efficacy of a tuberculosis DNA vaccine. AB - Tuberculosis is the most widespread and lethal infectious disease affecting humans. Immunization of mice with plasmid DNA constructs encoding one of the secreted components of Mycobacterium tuberculosis, antigen 85 (Ag85), induced substantial humoral and cell-mediated immune responses and conferred significant protection against challenge with live M. tuberculosis and M. bovis bacille Calmette-Guerin (BCG). These results indicate that immunization with DNA encoding a mycobacterial antigen provides an efficient and simple method for generating protective immunity and that this technique may be useful for defining the protective antigens of M. tuberculosis, leading to the development of a more effective vaccine. PMID- 8705860 TI - Suppressive vaccination with DNA encoding a variable region gene of the T-cell receptor prevents autoimmune encephalomyelitis and activates Th2 immunity. AB - A variable region gene of the T-cell receptor, V beta 8.2, is rearranged, and its product is expressed on pathogenic T cells that induce experimental autoimmune encephalomyelitis (EAE) in H-2u mice after immunization with myelin basic protein (MBP). Vaccination of these mice with naked DNA encoding V beta 8.2 protected mice from EAE. Analysis of T cells reacting to the pathogenic portion of the MBP molecule indicated that in the vaccinated mice there was a reduction in the Th1 cytokines interleukin-2 (IL-2) and interferon-gama. In parallel, there was an elevation in the production of IL-4, a Th2 cytokine associated with suppression of disease. A novel feature of DNA immunization for autoimmune disease, reversal of the autoimmune response from Th1 to Th2, may make this approach attractive for treatment of Th1-mediated diseases like multiple sclerosis, juvenile diabetes and rheumatoid arthritis. PMID- 8705861 TI - Large clonal expansions of CD8+ T cells in acute infectious mononucleosis. AB - Primary infection with Epstein-Barr virus often results in the clinical syndrome of acute infectious mononucleosis (glandular fever). This illness is characterized by a striking lymphocytosis, the nature of which has been controversial. We show that large monoclonal or oligoclonal populations of CD8+ T cells account for a significant proportion of the lymphocytosis and provide molecular evidence that these populations have been driven by antigen. The results suggest that the selective and massive expansion of a few dominant clones of CD8+ T cells is an important feature of the primary response to this virus. PMID- 8705862 TI - Alternatively spliced mdm2 transcripts with loss of p53 binding domain sequences: transforming ability and frequent detection in human cancer. AB - The mdm2 oncogene encodes a 90-kilodalton nuclear phosphoprotein that binds and inactivates the p53 tumor suppressor protein. Here we report the observation of five alternatively spliced mdm2 gene transcripts in a range of human cancers and their absence in normal tissues. Transfection of NIH 3T3 cells with each of these forms gave foci of morphologically transformed cells. A higher frequency of splice variants lacking p53 binding domain sequences was found in late-stage and high-grade ovarian and bladder carcinomas. Four of the splice variants show loss of p53 binding, consistent with partial deletion of sequences encoding the p53 binding domain, but retain carboxyterminal zinc-finger domains. These observations suggest a reassessment of the transforming mechanisms of mdm2 and its relation to p53. PMID- 8705863 TI - The seroepidemiology of human herpesvirus 8 (Kaposi's sarcoma-associated herpesvirus): distribution of infection in KS risk groups and evidence for sexual transmission. AB - Striking differences in Kaposi's sarcoma (KS) risk for AIDS patients who acquire HIV via homosexual activity and those whose HIV infections derive from blood product exposure suggest the presence of a sexually transmitted agent other than HIV in the development of KS. Using an immunofluorescence assay, we examined serum samples from 913 patients for the presence of antibody specific for infection by human herpesvirus 8 (HHV8), an agent whose genome is regularly found in KS tissue. The distribution of HHV8 seropositivity conforms to that expected for a sexually transmitted pathogen and tracks closely with the risk for KS development. Our data support the inference that this virus is the etiologic cofactor predicted by the epidemiology of KS. PMID- 8705864 TI - KSHV antibodies among Americans, Italians and Ugandans with and without Kaposi's sarcoma. AB - A major controversy regarding Kaposi's sarcoma-associated herpesvirus (KSHV or HHV8) is whether or not it is a ubiquitous infection of humans. Immunoassays based on KSHV- and Epstein-Barr virus (EBV)-coinfected cell lines show that most US AIDS-KS patients have specific antibodies to KSHV-related antigens. We have developed a sensitive indirect immunofluorescence assay (IFA) based on an EBV negative, KSHV-infected cell line, BCP-1. When we used this IFA assay, KSHV related antibodies were found in 71-88% of serum samples from US, Italian and Ugandan AIDS-KS patients, as well as all serum samples examined from HIV seronegative KS patients. Although none of the US blood donors examined were KSHV seropositive by IFA, intermediate and high seroprevalence rates were found in Italian and Ugandan control populations. Antibody kinetics showed that more than half of the AIDS-KS patients who were examined IgG-seroconverted before KS development, and antibody levels did not decline after seroconversion. For these patients, seropositivity rates increased linearly with time, suggesting that the rate of infection was constant and that the risk of developing KS once infected with KSHV is not highly dependent on the duration of infection. These data strongly suggest that KSHV is not ubiquitous in most populations and that the virus may be under strict immunologic control in healthy KSHV-infected persons. PMID- 8705865 TI - Enhancement of vowel coding for cochlear implants by addition of noise. AB - Profoundly deaf people, who gain no benefit from conventional hearing aids, can receive speech cues by direct electrical stimulation of the cochlear nerve. This is achieved by an electronic device, a cochlear implant, which is surgically inserted into the ear. Here we show physiological results from the isolated sciatic nerve of the toad Xenopus laevis, used to predict the response of the human cochlear nerve to vowels coded by a cochlear implant. These results suggest that standard analogue cochlear implants do not evoke the patterns of neural excitation that are normally associated with acoustic stimulation. Adding noise to the stimulus, however, enhanced distinguishing features of the vowel encoded by the fine time structure of neural discharges. On the basis of these results, and those concerning stochastic resonance, we advocate a cochlear implant coding strategy in which noise is deliberately added to cochlear implant signals. PMID- 8705866 TI - Applying very high resolution microfocus X-ray CT and 3-D reconstruction to the human auditory apparatus. AB - Conventional high-resolution X-ray computed tomography (XCT) is an important medical technique because it provides sectional images (tomograms) of internal structures without destroying the specimen. However, it is difficult to observe and to analyze fine structures less than a few cubic millimeters in size because of its low spatial resolution of 0.4 mm. Overcoming this problem would not only enable visualization of human anatomical structures in living subjects by means of computer images but would make it possible to obtain the equivalent of microscopic images by XCT without making microscopic sections of biopsy material, which would allow the examination of the entire body and detection of focal lesions at an early stage. Bonse et al. and Kinney et al. studied absorption contrast microtomography by using synchrotron radiation and achieved 8-microns spatial resolution in human cancellous bone. Recently, Momose et al. reported examining the soft tissue of cancerous rabbit liver by a modification of the phase-contrast technique using synchrotron radiation with a spatial resolution of 30 microns (ref. 4). However, the equipment for synchrotron radiation requires a great deal of space and is very expensive. Aoki et al., on a different tack, reported microtomography of frog embryos by using a conventional laboratory microfocus X-ray source with a spot size of about 2 microns (ref. 5). As no human tomographic studies by superresolution microfocus XCT (MFXCT) using a normal open type X-ray source have been reported, we tried using MFXCT with a maximum experimental spatial resolution of 2.5 microns, especially designed for industrial use, on the auditory ossicles of a human fetus, the smallest and lightest bones in the skeletal system. No XCT studies of fetal auditory ossicles have been reported to date. The fine tomograms with three-dimensional reconstructions obtained showed the existence of an apparently previously undescribed joint between the tympanic ring and the anterior process of the malleus. We hope the early development of this MFXCT for clinical use will make a great contribution to medicine. PMID- 8705867 TI - Biomedical three-dimensional holographic microimaging at visible, ultraviolet and X-ray wavelengths. AB - A new imaging technology is under advanced development that has several key advantages over conventional forms of microimaging performed with standard light microscopes, confocal light microscopes, and electron microscopes. The image created by this microscope possesses several unique features: It is intrinsically three-dimensional; it can be formed with very high contrast, a characteristic of the phase-sensitive nature of the recording; the information contained in the image is obtained in a single exposure of the specimen, a feature that eliminates the accumulation of damage to living systems that can occur with techniques utilizing multiple exposures; the method of image construction is fundamentally free from aberration (distortion), thereby obviating the need to employ complex procedures for correction; the exact focal plane of any optical section is digitally determined through computation and is not based on any mechanical adjustments; and the principles of operation, including the computational processes and modalities of image presentation, are uniform over the full range of spectral coverage spanning from the visible (approximately 500 nm) to the X ray (approximately 0.3 nm) regions. The application of this new technology is expected to open new cost-effective avenues to the understanding, prevention and treatment of broad areas of human disease. PMID- 8705868 TI - Semiconductor microlasers: a new approach to cell-structure analysis. PMID- 8705869 TI - The progression of myopia in school age children: data from the Columbia Medical Plan. AB - The purpose of this study was to characterize the clinical course of myopia in a selected pediatric population. The computerized records of a health maintenance organization provided longitudinal data on 501 children aged 5 to 15 years. We examined the influence of age at diagnosis, gender, race, and initial refraction on progression of myopia. Mean follow-up was 34 months, with 75% of children having follow-up longer than 16 months. The mean rate of myopic progression was greater for children whose myopia was diagnosed at a younger age (5-7 years, 0.56 diopters/year versus 11-15 years, -0.28 diopters/year; p < 0.0001). Children with more than one diopter of myopia at first diagnosis progressed faster than children with less than or equal to one diopter of myopia (mean rate -0.48 diopter/year versus -0.41 diopter/year; p = 0.05). Cumulative event rate curves suggest a gender effect in the prepuberty years of 8 to 10, with myopia progressing faster in girls than boys (p = 0.003). Progression of myopia did not differ between white and non-white children. More rapidly progression of myopia is associated with younger age at initial diagnosis and greater severity of initial myopic refraction. PMID- 8705870 TI - Prevalence and causes of low vision and blindness in the Extreme North Province of Cameroon, West Africa. AB - A survey to determine the prevalence and causes of blindness and visual impairment in the Extreme North Province of Cameroon was conducted in the Spring of 1992. A total of 10,647 people age 6 years and older was selected from a multi stage, clustered sample stratified by ecological zone. The subjects were examined by ophthalmologist-led teams for visual acuity and ocular diseases. Approximately 1.2% of the sample was bilaterally blind by the World Health Organization classification (Category 3) of vision less than the ability to count fingers at 3 meters. Similarly to results found in other developing countries, senile cataract was the most common diagnosis encountered and the most frequent principal cause of low vision and blindness. PMID- 8705871 TI - Review of cataract intervention in Maharashtra State. AB - Blindness due to cataracts presents an enormous problem, not only in terms of human suffering, but also in terms of economic loss and social burden. In the absence of proven measures of primary prevention, secondary prevention in the form of surgical intervention is, currently, the remedy to the problem of blindness due to cataracts. There are various therapeutic approaches in cataract extraction. However, the conventional ICCE/SPECT and the newer ECCE/IOL are the popular approaches presently adopted on a mass scale. Maharashtra is consistently achieving the quantitative targets for cataract surgery given by the Government of India. However, the qualitative aspects of cataract surgery, the desired outcome of which is sight restoration in those blinded by cataracts, has hitherto not been monitored and assessed. The Kolhapur Circle was selected for this purpose. With the improvement of Ophthalmic Surgeons and PMOAS, the sample of cataract patients operated on in the year 1992-93 was followed. Two blocks from each of three districts in the circle were selected at random. The respondents were patients from these six blocks who had cataract surgery in the year 1992-93. 50% of the operations took place in the mobile camps while 20% took place in government fixed facilities and 30% in the private sector. Micro-analysis of the quantitative performance of cataract surgery in the Kolhapur Circle revealed that the circle is continuously delivering the intended benefits, i.e. relieving human suffering due to blindness from cataracts. The overall success rate of cataract surgery was 86% (Postoperative VA > 6/60). It seems that the investment made in cataract intervention is highly productive. PMID- 8705872 TI - Retinoblastoma referral patterns in Saudi Arabia. AB - The purpose of this study was to review the patterns of referral and presentation of patients with retinoblastoma in Saudi Arabia from 1983-1994. Retinoblastoma represents a particularly high volume at the King Khaled Eye Specialist Hospital and the Hospital's Retinoblastoma Registry provided relevant data. Records of 257 registered patients were analyzed. Of these, 112 were bilateral whereas 145 were unilateral, resulting in a total of 369 eyes. Referral diagnosis accuracy was high, 75% of referring physicians ranking retinoblastoma at the top of their differential diagnosis. Delay in referral was found to average 9.4 weeks. However, 49.4% of eyes were staged at Reese-Ellsworth Stage V at presentation. Significantly, the number of patients presenting with extraocular disease fell from 22.8% (1983-1988) to 12.3% (1989-1994), highlighting increasing awareness of retinoblastoma and availability of services in Saudi Arabia. PMID- 8705873 TI - Visual sensitivity and age-related eye diseases. The Beaver Dam Eye Study. AB - OBJECTIVE: To evaluate the relationships between visual sensitivity and age related ocular disorders. DESIGN: Prevalence survey of age-related eye diseases. SETTING: Beaver Dam, Wisconsin. PARTICIPANTS: A total community sample of persons 43-84 at the time of the census (1987-1988) was invited to participate (n = 5.926). MAIN OUTCOME MEASURE: Visual sensitivity was measured by a perimeter. Sensitivities were evaluated with regard to age, sex and age-related eye diseases. RESULTS: Sensitivity was inversely associated with age and was lower in women in each age stratum. Central cataract, cataract surgery, and late age related maculopathy were significantly related to decreased sensitivity in multivariate analyses. CONCLUSIONS: Visual sensitivity, an indication of the ability of the eye to detect a light stimulus, was diminished in those with central cataract, cataract surgery, and late age-related maculopathy. This may be related to diminished vision in environmental conditions of low illumination. Since visual sensitivity is relative easy to determine, this test may have use as a surrogate to assess functional vision loss. Also, although cataract surgery often leads to improved high contrast acuity, there may be a loss in low contrast settings. PMID- 8705874 TI - Three novel and one C31133T (Arg-338-->Stop) mutations of antihemophilic factor IX gene detected in Taiwan. AB - BACKGROUND: Hemophilia-B is caused by mutation of the coagulation factor IX (F.IX) gene. The gene is about 34 kilobases in length and contains 8 exons ranging from 25 to 548 base pairs. There is no common mutation pattern of this gene in any population as there is in thalassemia. The hope is to localize the mutations in patients for use in making rapid familial surveys and for prenatal diagnoses. METHODS: Diagnoses for a male patient with a newly-discovered mutation in Taiwan and three others with different novel mutations of F.IX gene were made from the results of prothrombin time (PT), activated partial thromboplastin time (APTT), assay of coagulation factors activity and factor IX antigen (F.IX:Ag) and also by assay of its inhibitor. The polymerase-chain-reaction (PCR)-based direct sequencing method was applied to evaluate all of the 8 exons and their flanking regions of F.IX gene for these patients. RESULTS: The four male patients are all severe hemophilia-B. In the three patients with novel mutation of F.IX gene, two have a very low F.IX:Ag of less than 1% and one has 73%, but no one has any inhibitor. One of the four mutations is A to C transversion at nucleotide (nt) 10458 resulting in tyrosine (69) to serine change in epidermal growth factor (EGF) type B domain. Although the F.IX:Ag is present, the F.IX activity (F.IX:C) is markedly deficit owing to the change of beta-hydroxylase recognition site. Another two have mutations at the splicing junctions. They are G to C transversion at nt 118, i.e. the first base of IVS-1 splicing donor, and G to T transversion at nt 17668, the last base of IVS-4 splicing recipient. These mutations may cause splicing failure and subsequently result in deficiency of both F.IX activity and antigen. The other mutation that has never previously presented in this country is a C to T transition (arginine-338 to stop) at nt 31133 that results in early termination of translation. CONCLUSIONS: The G to C mutation at nt 118 creates an Alu I restriction site and the C to T mutation at nt 31133 abolishes a Taq I site. Therefore, familial survey and rapid prenatal diagnosis for these patients by PCR-based direct sequencing method or by the restriction of enzyme cutting method pointing at the known mutation sites are certainly possible to achieve in our patients. PMID- 8705875 TI - Allogeneic bone marrow transplantation for fourteen patients with severe aplastic anemia. AB - BACKGROUND: Bone marrow transplantation (BMT) is the best curative approach for younger patients with severe aplastic anemia (SAA). Major obstacles to success of allogeneic BMT include graft-versus-host disease (GVHD), graft rejection and treatment related toxicities. Experience with 14 SAA patients who received BMT is reported here. METHODS: From December 1986 to May 1995, 14 patients with SAA were treated with BMT; 13 were allogeneic, and 1 was syngeneic. There were nine males and five females whose average age was 24.7 years (range 15-36 years). The median pretransplant disease duration was 93 days (range 7-610 days). Five patients were nontransfused before BMT. The pretransplant conditioning regimen consisted of 200 mg/kg cyclophosphamide (CY) intravenously, divided over four consecutive days, followed by 300 cGy total-body irradiation (TBI) on the day before BMT. Two untransfused, one transfused patient and one syngeneic transplant received CY only as preconditioning. For GVHD prophylaxis, the 13 patients were given a combination of cyclosporine and a short course of methotrexate. RESULTS: Of the 14 patients, 11 were still alive 10 to 90 months later, with functional engraftment; the median survival of 39 months. There were three deaths including one with primary graft failure with intracranial hemorrhage, and two with delayed graft rejection and sepsis. The patient who received syngeneic BMT developed late graft failure six months post-transplant, but was successfully treated with a second BMT. Acute GVHD occurred among 5 of the 13 engrafted patients, only one of whom was Grade III clinically. Chronic GVHD was observed in 2 out of 10 evaluable patients. CONCLUSIONS: The combination of CY and TBI is an effective, well tolerated conditioning regimen for BMT in patients with SAA. The acute GVHD rate was low in our patients receiving cyclosporine. BMT is the treatment-of-choice for patients under the age of 40 with SAA, for those with human leucocyte antigen (HLA)-identical siblings or an identical twin and particularly for those patients who have not received transfusion. PMID- 8705876 TI - Characteristics of endoscopic retrograde cholangiopancreatogram in patients with primary biliary cirrhosis. AB - BACKGROUND: The findings of cholangiopancreatograms in patients with primary biliary cirrhosis vary markedly in literature. We tried to evaluate the changes of endoscopic retrograde cholangiopancreatograms in Chinese patients with primary biliary cirrhosis. METHODS: Twenty-nine patients with primary biliary cirrhosis underwent endoscopic retrograde cholangiopancreatography over the past 8 years. Three patients with a negative antimitochondrial antibody test were excluded. Well opacified cholangiograms and pancreatograms were obtained in 24 patients. Meanwhile, 16 subjects with normal cholangiopancreatogram served as controls. The characteristics and prevalence of abnormal cholangiopancreatograms in the patients and the correlation of radiography with clinical severity of the disease were evaluated. RESULTS: The maximum diameters of the common bile duct (9.7 +/- 4.0 vs. 7.6 +/- 0.9 mm, NS), right (5.0 +/- 1.6 vs. 4.4 +/- 1.2 mm, NS) and left (5.1 +/- 1.2 vs. 4.9 +/- 1.4 mm, NS) intrahepatic ducts did not show significant difference between the patients with primary biliary cirrhosis and the controls. Abnormal intrahepatic cholangiograms were obtained in 12 (50%) patients including eight with diminished arborization and focal stenosis, three with crowding and tortuous branches and one with focal stenosis alone. A hepatic hilum notch on the common hepatic duct was found in eight (33.3%) patients. The abnormalities of intrahepatic ducts did not correlate with age, sex, Pugh's scores, various liver function tests or histologically cirrhotic change. One (4.2%) patient had an abnormal pancreatogram. CONCLUSIONS: Abnormal intrahepatic cholangiograms are present in half of patients with primary biliary cirrhosis, but are not related to clinical severity. PMID- 8705877 TI - Relationship between C14 urea breath test values and solid-phase gastric emptying time in non-ulcer dyspepsia patients. AB - BACKGROUND: Non-ulcer dyspepsia (NUD) is a poorly understood syndrome often found with endoscopic evidence of gastritis; Helicobacter pylori (Hp) is a common and important cause of gastritis. In the recent literature, gastric hypomotility is thought to be a cause of NUD. Thus, this investigation studied the relationship between Hp and delayed gastric emptying in NUD patients. METHODS: Using a radionuclide-labelled solid meal to calculate gastric emptying time (GET) of 78 NUD patients. The carbon-14 urea breath test (C14 UBT) was used to quantitate Hp infection. RESULTS: The prevalence of Hp infection in patients with NUD reached 59%. There was a strong association of Hp infection with advanced age (p = 0.0091). There was no significant difference between solid-phase GET and C14 UBT values among three different age groups (young, middle, old) of NUD patients. There was no difference among sex, age, body weight and solid-phase GET between Hp-positive and Hp-negative NUD patients. However the solid-phase GET was significantly prolonged in patients with NUD, compared with the controls. CONCLUSIONS: Solid-phase GET is not correlated with the C14 UBT values, and Hp gastric colonization does not account for dyspeptic syndrome in NUD patients. PMID- 8705878 TI - Dynamic infusion cavernosometry and cavernosography (DICC) in the evaluation of vasculogenic impotence. AB - BACKGROUND: This study was conducted to find more accurate physiological assessment of the penile vascular status in impotent patients. METHODS: Twenty two males, aged 25-76 years (mean 54.0 years) with possible underlying vascular causes of impotence, had dynamic infusion cavernosometry and cavernosography (DICC) examination to ascertain the arterial and/or venous insufficiency in the penis. Prostaglandin E1 20 micrograms was injected intracavernously in this 4 phase study. RESULTS: No side effect or complication was noted during or after the examination in all 22 patients. During the phase 1 test, the mean resting intracavernous pressure (ICP) was 6.8 +/- 3.6 mmHg and the mean plateau ICP after PGE1 injection was 47.2 +/- 23.9 mmHg. The mean ratio of plateau ICP to mean arterial pressure was 50.2 +/- 23.4%. In the phase 2, the mean ICP determined 30 seconds after cessation of saline infusion without and with perineal compression was 65.3 +/- 26.8 mmHg and 129.1 +/- 26.2 mmHg, respectively. The mean ICP decay from 150 mmHg was 84.7 +/- 26.8 mmHg. The mean systolic pressure of each cavernous artery, 87.6 +/- 18.9 mmHg and 93.6 +/- 17.6 mmHg in left and right side, respectively, was found during phase 3 evaluation. The mean ratio of systolic pressure of each cavernous artery to brachial artery (cavernous-brachial index, CBI) was 70.1 +/- 12.7% and 75.1 +/- 11.5% in left and right side, respectively. Six patients with CBI greater than 75% had normal Doppler duplex sonography (diameter of cavernous artery greater than 0.9 mm and peak flow velocity greater than 25 cm/sec). Cavernosography in phase 4 evaluation showed venous leakage in 20 of 22 patients (90.9%) with ICP decay greater than 60 mmHg. Two patients with ICP decay less than 45 mmHg were found to have no venous leakage. CONCLUSIONS: The cavernous-brachial index is usually less than 75% in penile arterial insufficiency. The patients with ICP decay from 150 mmHg greater than 60 mmHg would have penile venous leakage demonstrated in the cavernosography. The results of cavernosometry are highly correlated with cavernosography. Our results suggest that DICC may provide a good examination for more physiological assessment and accurate diagnosis of vasculogenic impotence. PMID- 8705879 TI - Treatment of open tibial fracture with Ender nail. AB - BACKGROUND: This retrospective study aims to evaluate the effect of Ender nail in management of open type I and II unstable tibial fractures. METHODS: From 1984 to 1993, 30 cases of open unstable tibial shaft fracture (with fibular fracture) were collected in the retrospective study. All were fixed with Ender nails. The followup period was 99 (24-135) months. RESULTS: The average of blood loss was 140 cc, operation time 45 minutes, hospital day 10 days, and union time 20.5 weeks. Student t-test revealed statistically significant difference between open type I cases and type II cases in some parameters described above. CONCLUSIONS: Ender nail is effective in the treatment of open type I and II unstable tibial shaft fractures, but it should be used carefully, especially in cases with more comminution. PMID- 8705880 TI - Leiomyosarcoma of the rectum: a series of twelve cases. AB - BACKGROUND: Leiomyosarcoma of the rectum is a rare disease without well documented report on its management and prognosis in Taiwan. To evaluate the effectiveness and prognosis of different treatments for rectal leiomyosarcoma, we reviewed a series of 12 patients with rectal leiomyosarcoma treated at our hospital. METHODS: This study is a retrospective review of 12 rectal leiomyosarcomas treated at VGH-Taipei from 1976 to 1995, with emphasis on symptoms, therapeutic procedures and prognosis. RESULTS: The majority of tumors occurred in men (n = 10) and the average age at presentation was 54.4 years. Bloody stool and perianal pain were the two most common symptoms. Nine tumors were found in the lower rectum, within 4 cm above the dentate line. Five patients received abdominoperineal resection, four received wide local excision, and three had diversion colostomy. The effect of adjuvant therapy, either chemo- or radiotherapy, was poor. The overall one-year and five-year survival rates were 83% and 46%, respectively. Liver was the principal metastatic site. The local recurrence rate was much higher in patients receiving wide local excision than in those receiving abdominoperineal resection (100% vs. 20%). CONCLUSIONS: Abdominoperineal resection is an appropriate choice of treatment for patients with operable rectal leiomyosarcoma due to its lower local recurrence rate. Wide local excision could be considered when the lesion is less than 2 cm in diameter or when patients are not suitable for radical surgery. The effect of adjuvant therapy, either radiotherapy or chemotherapy, is poor for rectal leiomyosarcoma. PMID- 8705881 TI - Creutzfeldt-Jakob disease: clinical analysis of eight cases. AB - BACKGROUND: Creutzfeldt-Jakob disease (CJD) or spongiform encephalopathy in humans, is a rare, rapidly progressive and fatal disorder of the central nervous system. Since clinical diagnosis is challenging, this retrospective investigation was performed. METHODS: Eight cases who had been diagnosed as CJD from 1987 to 1995 were reviewed thoroughly, with a resulting clinical diagnosis of CJD. Two cases were proved by brain biopsy. RESULTS: Four males and four females were included; their ages were 53 to 81 years with a mean of 66 years. The mean illness duration of the six patients who died was six and a half months. Clinical findings were variable and included dementia, aphasia, ataxic gait, pyramidal, extrapyramidal, myoclonus and visual problems. CONCLUSIONS: For diagnosis of CJD, clinical suspicion is very important. Electroencephalogram (EEG) remains the most helpful laboratory diagnostic tool; serial recordings are necessary if the initial EEG finding is nonspecific. Single photon emission computed tomography (SPECT) may be helpful for premorten diagnosis of CJD, but sensitivity and specificity need further investigation. Brain biopsy seemed to be unnecessary if the clinical, EEG and SPECT findings are typical. PMID- 8705882 TI - Cytomegalovirus colitis in an immunocompetent old woman successfully treated with ganciclovir: a case report. AB - Cytomegalovirus (CMV) diseases occur almost exclusively in the immunocompromised hosts. Persons most commonly affected are patients with acquired immunodeficiency syndrome (AIDS). On rare occasions, however, CMV diseases can be seen in apparently immunocompetent persons. A CMV colitis in a 68-year old immunocompetent woman presenting with watery diarrhea, malaise, and body weight loss of about 5 kg over a three week period is reported. Colonoscopy and mucosal biopsy revealed CMV colitis involving the sigmoid colon. Treatment for two weeks with ganciclovir (10 gm/kg/day I.V. in 2 divided doses) resulted in resolution of colitis and clinical symptoms without any noticeable side-effects. There was no relapse of infection by six months of follow-up. The possibility of CMV colitis should be considered in any elderly person with watery diarrhea, general debilitation, marked body weight loss and with negative stool cultures. CMV colitis may be more frequent than is usually believed. It has a favorable response to ganciclovir without further relapse after the successful treatment in patients with normal immune function. PMID- 8705883 TI - Watershed infarction of spinal cord after the embolization of bronchial artery: a case report. AB - A case of watershed infarction in the spinal cord is reported. The patient underwent bronchial artery embolization for control of massive hemoptysis. The bronchial arteriogram was carefully examined and focused on blood supply to the spinal cord prior to embolization. Acute paraparesis followed the embolization procedure even though there was no visible spinal supply on the arteriogram. Magnetic resonance imaging showed a hyperintensity lesion over the watershed region which is located at the central portion of the upper thoracic cord. This case is reported to emphasize the significant role which angiographically invisible small vessels can play in the blood supply to the spinal cord. The vascular system of the spinal cord and the prevention of spinal cord ischemia secondary to embolization are further discussed here. PMID- 8705884 TI - Pheochromocytoma of the urinary bladder: a case report. AB - A 61-year-old female was admitted who had had symptoms of hematuria and a tumor mass in the urinary bladder for six months. Hypertension and headache after micturition had been noted for one year. Plasma norepinephrine before and after voiding were elevated. Increased I-131 metaiodobenzylguanidine (I-131 MIBG) uptake in the urinary bladder was also found. The plasma norepinephrine level and blood pressure returned to normal after partial cystectomy. Diagnosis of pheochromocytoma was confirmed by histology. PMID- 8705885 TI - Alcaligenes xylosoxidans neonatal meningitis: a case report. AB - Neonatal meningitis caused by Alcaligenes xylosoxidans is associated with a high mortality rate. The causative microorganism is resistant to most antimicrobials. Generally, once the organism has been isolated from the cerebrospinal fluid of the infected neonate, initial therapy with a third-generation cephalosporin and trimethoprim-sulfamethoxazole is recommended before determining its susceptibility to antimicrobials. There present this is the first of neonatal meningitis with transient diabetes insipidus cause by A. xylosoxidans. The patient was treated with dDAVP for seven days and a combination of imipenem plus trimethoprim-sulfamethoxazole for 28 days. The patient was discharged in a stable condition and the end of that time, but with sequelae of hydrocephalus and hearing impairment. PMID- 8705886 TI - Human papillomavirus in cyclophosphamide and diverticulum-associated squamous cell carcinoma of urinary bladder: a case report. AB - The association of cyclophosphamide therapy with the development of urothelial malignancy has been documented. Bladder diverticula associated with squamous cell carcinoma were also identified. In addition, incorporation of human papillomavirus (HPV) deoxyribonucleic acid (DNA) has been previously shown in urothelial carcinoma by in situ hybridization technique. Here a case is reported of bladder squamous cell carcinoma occurring four years after intravenous pulse therapy with cyclophosphamide in a patient with bladder diverticula where HPV was identified in the lesion. To present knowledge, this is the first case report of bladder squamous cell carcinoma with simultaneous occurrence of three risk factors for urothelial malignancy: cyclophosphamide therapy, diverticulum, and HPV infection. PMID- 8705887 TI - Cytomegalovirus retinitis after heart transplant: a case report. AB - A man aged 51 years was found to have cytomegalovirus (CMV) retinitis in his right eye one year after heart transplant surgery. Ophthalmoscopy revealed a patch yellowish-white, granular, necrotic lesions admixed with flame-shaped intraretinal hemorrhage over the temporal lower retina. Anti-CMV IgM antibody was positive. The visual acuity improved from 20/200 to 20/20 within one month by CMV hyperimmune globulin therapy (cytotect, Biotest, FRG, 2 ml/kg every other day for two weeks). The retinal necrosis healed with permanent scar tissue formation. No other complications were noted after a fifteen month follow up. In this study, the clinical findings and fundus picture were presented, along with previous literature reviewed. PMID- 8705888 TI - Teaching human anatomy to medical students. PMID- 8705889 TI - The most restrictive alternative. PMID- 8705890 TI - Trends and patterns in health insurance coverage, 1991-1994. PMID- 8705891 TI - Evolutionary morphology at Brown: what's it all about? PMID- 8705893 TI - Navel encounters. PMID- 8705892 TI - Anatomy and more. PMID- 8705894 TI - TennCare BHOs. Partners program startup mirrors TennCare; mental health patients lumped into two plans. PMID- 8705895 TI - Where did all the specialties go? Managed care trends steer tomorrow's MDs into primary care slots. PMID- 8705896 TI - Loss prevention case of the month. Difficult diagnosis. PMID- 8705897 TI - Cardiac tamponade as the initial presentation of bronchioloalveolar cell carcinoma. AB - Bronchioloalveolar cell carcinoma (BCC) is a relatively uncommon form of bronchogenic cancer with an insidious onset and a variety of radiographic appearances. Although the World Health Organization currently classifies it as a subtype of adenocarcinoma, many investigators view BCC as a distinct tumor with characteristic clinical features. There are still uncertainties regarding its origin, as well as the significance of associated clinical findings such as cardiac tamponade. PMID- 8705898 TI - Repair of penetrating cardiac injury using skin staples. AB - Cardiac stapling is a highly effective technique in the management of hemorrhage from penetrating cardiac injuries. It may allow the salvage of patients with multiple cardiac lacerations who would not otherwise survive following standard suture techniques for repair. Cardiac stapling is probably not indicated in complex injury cases such as those from gunshot, and the trauma surgeon must use judgment in applying the staple technique, though its use for cardiography in the ED and the OR will minimize the risk of contamination of personnel from a needle stick from the repair portion of the surgical procedure. Staplers are readily available, easy to use and safe to surgical personnel, and they provide rapid and effective hemostasis. PMID- 8705899 TI - A diabetic man with left shoulder pain. PMID- 8705900 TI - Postexposure immunoprophylaxis and chemoprophylaxis for communicable diseases. PMID- 8705901 TI - Triple-marker screening for Down syndrome. PMID- 8705902 TI - Triple-marker screening for Down syndrome. PMID- 8705903 TI - Health care the tie that binds. PMID- 8705904 TI - Blood recycling eliminates need for blood. PMID- 8705905 TI - DNR orders in stroke. PMID- 8705906 TI - The NCIC-Manitoba Breast Tumor Bank: a resource for applied cancer research. AB - The NCIC-Manitoba Breast Tumor Bank is one of several tumour banks that have been established through the Molecular Epidemiology Program of the National Cancer Institute of Canada (NCIC). The NCIC-Manitoba Breast Tumor Bank is an example of one model developed to facilitate research designed to translate the findings of basic science into information useful in the clinical arena. The tumour bank's mandate is to provide a national resource that consists of a preassembled dataset of matched samples of paraffin-embedded and frozen tumour tissue with corresponding pathological and clinical data. In the first 3 years the tumour bank has accrued data and samples from over 1800 cases of breast cancer and has provided support for 20 research projects across Canada and the United States. PMID- 8705907 TI - Prevalence of factor V Leiden in a Canadian blood donor population. AB - OBJECTIVE: To determine the prevalence of factor V Leiden in a Canadian blood donor population. DESIGN: Cross-sectional laboratory study. SETTING: Hamilton Centre of the Canadian Red Cross Society. PARTICIPANTS: Volunteer donors who attended Hamilton Centre blood donor clinics over a 4-day period in August 1994; blood samples from 356 people were evaluable. OUTCOME MEASURES: Presence of factor V Leiden. RESULTS: Factor V Leiden was detected in 19 of the 356 people, for a prevalence rate of 5.3% (95% confidence interval 3.0% to 7.6%). All 19 people were shown to be heterozygous for the mutation. CONCLUSION: Factor V Leiden is common in the Canadian population. Its prevalence is similar to that reported in other Western countries. These data are relevant in the clinical management of patients at risk for venous thrombosis and those with recurrent thrombotic disorders. PMID- 8705908 TI - Elevated serum digoxin levels in a patient taking digoxin and Siberian ginseng. AB - A 74-year-old man taking a constant dose of digoxin for many years was found to have an elevated serum digoxin level with no signs of toxic effects. Common causes of elevated serum digoxin were ruled out, and the patient's digoxin level remained high after digoxin therapy was stopped. The patient then revealed that he was taking Siberian ginseng, a popular herbal remedy. The patient stopped taking ginseng, and the serum digoxin level soon returned to an acceptable level. The digoxin therapy was resumed. The patient resumed taking ginseng several months later, and the serum digoxin level again rose. Digoxin therapy was maintained at a constant daily dose, the ginseng was stopped once more, and the serum digoxin levels again returned to within the therapeutic range. It is unclear whether some component of the ginseng was converted to digoxin in vivo, interfered with digoxin elimination or caused a false serum assay result. The author cautions physicians to be alert to the potential for herbal remedies to interact with prescribed medications and to affect biochemical analyses. PMID- 8705909 TI - Factor V Leiden: who should be tested? AB - Resistance to activated protein C resulting from the genetic point mutation known as factor V Leiden is the most frequently found genetic risk factor associated with familial predisposition to venous thrombosis. Factor V Leiden is also frequent among people with nonfamilial venous thrombosis and appears to have a relatively high prevalence rate in the general population. The author comments on the findings of the first Canadian prevalence study of factor V Leiden, reported in this issue by Dr. David H. Lee and associates (see pages 285 to 289). She notes that although certain hereditary and clinical variables are known to modulate the risk of venous thrombosis in people with factor V Leiden, explanations for the relatively high prevalence of this mutation and the wide spectrum of risk associated with it are still speculative. Management guidelines for affected patients are quickly evolving but are still limited by a lack of clinical data. It is clear that further research into factor V Leiden will have considerable importance for the understanding and management of thrombotic risk. PMID- 8705910 TI - Funding medical and health-related research in the public interest. AB - Public funding for medical and health-related research in Canada is declining. At the same time, the pharmaceutical industry is directing increasing amounts of money to publicly funded agencies such as universities and the Medical Research Council of Canada. However, the kinds of research most valuable to commercial firms may not be those most valuable to the Canadian public. There is a danger that research priorities and activities in public institutions may become skewed as a result of increased drug-industry funding. Mechanisms need to be found to ensure an appropriate balance between the research that is most valuable to the public interest and to the long-term advancement of knowledge, and the research that is likely to lead to marketable products. One such mechanism is the direction of a proportion of the money from drug companies to a "no-strings attached" fund specifically to support types of research that are in the public interest but not likely to lead to marketable products. PMID- 8705911 TI - Supplementary statement on hepatitis A prevention. Laboratory Centre for Disease Control. PMID- 8705912 TI - After 4 years' work, revised Code of Ethics goes to General Council for decision. AB - The CMA's Committee on Ethics will present a revised Code of Ethics for consideration by General Council during the annual meeting in Sydney, NS, later this month. This article outlines the reasons for updating the current (1990) version of the code and explains some of the significant changes and omissions. If approved by General Council, the revised code will take effect immediately. PMID- 8705914 TI - After slow start, project to channel drug company funds to universities builds steam. AB - Drug companies appear to have been listening when researchers began complaining about their lack of participation in the MRC/PMAC Health Program, for the 30 month-old project appears to have taken on new life. It is designed to increase collaboration between university and pharmaceutical industry researchers by directing more of the industry's growing investment in Canada through the MRC's peer-review process. By mid-May, program commitments stood at $60 million. PMID- 8705913 TI - CMA's Clinical Q&A discussion group offers corridor consultations on the Internet. AB - A CMA-sponsored Internet discussion group, Clinical Q&A, has become a popular feature of CMA Online. About 150 physicians from around the world have participated in discussions and question-answer interaction, usually about diagnostic or patient-management dilemmas. Based on the success of the venture, CMA Online plans to expand the Internet forums it offers to CMA members. PMID- 8705915 TI - Healing all the king's men: medical treatment at Fortress Louisbourg, 1744. AB - When the CMA General Council meets in Cape Breton, NS, Aug. 18-21, physicians will have an opportunity to visit the largest reconstructed colonial town in North America, which is only 40 km from the meeting site in Sydney. This article looks at the medical treatment available at the historic Fortress of Louisbourg in 1744, when it was France's Atlantic coast bastion. PMID- 8705916 TI - Women's health research assumes higher profile at two Toronto teaching hospitals. AB - Two University of Toronto teaching hospitals have established chairs in women's health. Dr. Donna Stewart, who was appointed to the Lillian Love Chair in Women's Health at the Toronto Hospital, says the initiatives highlight the growing importance of women's health issues. Dr. Anna Day, physician-in-chief at Women's College Hospital, which established chairs in breast cancer and women's health research, hopes that a way will be found to coordinate women's health programs to avoid duplication of research efforts. PMID- 8705917 TI - HIV/AIDS in the context of culture: selection of ethnocultural communities for study in Canada. AB - This article reports on the methodology used to select six ethnocultural communities invited to participate in subsequent phases of the project on HIV/AIDS in the context of culture in Canada. Selection was based on quantitative data on demography, qualitative assessment of ethnocultural cohesion; and quantitative data and qualitative data of exposure to risk for sexually transmitted disease. A principle of partnership insured that the final selection was completed by interaction between the investigators and the National Advisory Committee representing ethnocultural communities in Canada. The six communities asked to participate in Phase II of the study were: in Montreal, the Latin American and the Arabic-speaking communities; in Toronto, the English-speaking Caribbean communities and communities from the Horn of Africa; in Vancouver, the Chinese and the South Asian communities. The results are significant for the future both of research on ethnicity in Canada and of control of HIV and AIDS. PMID- 8705918 TI - Participatory aspects in the qualitative research design of phase II of the ethnocultural communities facing AIDS study. AB - This paper describes the steps taken in generating and implementing a qualitative research design for Phase II of the Ethnocultural Communities Facing AIDS Study. Theoretically framed by the macro-level, sociocultural model of health behaviour developed by Kleinman, the methodological procedures are an adaptation of Scrimshaw's Rapid Assessment Procedures (RAP) and a participatory approach involving stakeholders from each ethnocultural community. Qualitative data-on behaviours conducive to HIV transmission in six ethnocultural communities in Canada-were elicited using a combination of key communicator interviews, focus groups, and participant observation techniques. Data were analyzed using systematic content analysis techniques. Inter-rater reliability checks and procedures of triangulation demonstrated the validity of evidence generated. A commitment to research partnership with community persons, and an accountability loop that provided assurances of how the data would be scientifically represented, were critical elements in the process of design construction. PMID- 8705920 TI - HIV/AIDS in the context of culture. PMID- 8705919 TI - Many voices--sociocultural results of the ethnocultural communities facing AIDS study in Canada. AB - This paper presents the results of Phase II of the Ethnocultural Communities Facing AIDS Study, the sociocultural investigation of factors contributing to risk behaviour associated with HIV/AIDS in six ethnocultural communities in Canada in three urban sites. In Vancouver, the South Asian and Chinese communities were studied, the Horn of Africa and English-speaking Caribbean communities in Toronto and the Latin American and Arabic-speaking communities in Montreal. Results demonstrated that there are common elements across these ethnocultural communities that increase the risk for HIV transmission. HIV/AIDS awareness and prevention in ethnocultural communities must address sociocultural differences, particularly sex role differences between men and women in terms of power within relationships to negotiate for safer sexual practices. PMID- 8705921 TI - Understanding use of condoms among Canadian ethnocultural communities: methods and main findings of the survey. AB - The aim of this study was to understand the intention to use a condom for each instance of sexual intercourse with a new partner in three of Canada's non dominant ethnocultural communities: Latin American (N = 346), English-speaking Caribbean (N = 358), and South Asian (N = 355). All respondents were recruited from multiple ethnocultural venues using predetermined sampling frames and quotas for each community. Anonymous questionnaires assessing culturally specific theoretical constructs were completed. This paper presents the methodology and the main findings. The high quality of the results of this study demonstrate the advantage of establishing strong partnerships with members of communities being studied. PMID- 8705922 TI - Canadian ethnocultural communities facing AIDS: overview and summary of survey results from phase III. AB - This paper describes the survey results reporting demographic profiles, behaviours, opinions, beliefs, attitudes, and intentions related to condom use for three Canadian ethnocultural communities (Latin American, English-speaking Caribbean and South Asian) participating in the Ethnocultural Communities Facing AIDS Study. Specific recommendations are presented for HIV-prevention programming based on the research results. PMID- 8705924 TI - Five conversations: reflections of stakeholders on the impact of the ethnocultural communities facing AIDS study. AB - This paper reports on issues identified in conversations held between one of the Ethnocultural Communities Facing AIDS Study (ECFA) investigators and five stakeholders associated with this experiment in collaborative and participatory research. The stakeholders reflect on their experiences in partnering with university-based researchers, resistance in ethnocultural communities to being researched, and the next steps implicated by the research in question. The problem of HIV/AIDS in ethnocultural communities in Canada is, relative to the mainstream, also viewed as a symbol of disadvantage in these communities. The stakeholders suggest that to address these deeper concerns, there must be vital, dynamic, and enduring communication between researchers, community representatives, and government at all levels, in the process of identifying solutions and implementing them in the present. PMID- 8705923 TI - Overview of the Canadian study on the determinants of ethnoculturally specific behaviours related to HIV/AIDS. AB - Canada's population is composed of heterogenous ethnocultural communities. There is a need for information and educational initiatives on HIV and AIDS directed specifically at these communities. For such interventions to be effective we must determine the existing personal and sociocultural factors related to HIV transmission. There has been little such research in Canada. In this supplement we report on various aspects of a study conducted between May 1992 and December 1994 to determine the factors related to HIV transmission in several ethnocultural communities. This paper describes some innovative aspects of the project: the conceptual framework, the community participatory model, the use of a multi-method research design, and the ongoing communication strategy. The combination of these elements makes the study unique. The value of the study lies not only in the information obtained but also in the model it provides for future research in other settings. PMID- 8705925 TI - Combining the energy of CNSs. PMID- 8705926 TI - Everyday functioning in Alzheimer's disease: contribution of neuropsychological testing. AB - COGNITIVE AND BEHAVIORAL problems associated with Alzheimer's disease are variable and complex; consequently, each patient should be managed by an advanced practice nurse (APN) so that everyday functioning is maximized and excess disabilities are prevented. Neuropsychological testing is sensitive to neuropathological conditions and holds promise for assisting APNs in the care of the Alzheimer's patient by pinpointing areas of cognitive and behavioral impairment. In this article, an overview is given of the essentials of a neuropsychological assessment for the Alzheimer's patient as performed by the neuropsychologist. Use of assessment results by the APN for designing interventions that compensate for losses but which preserve remaining abilities is discussed. PMID- 8705927 TI - Coming out: an overlooked concept. AB - WITH CURRENT TRENDS in human immunodeficiency virus (HIV) early intervention moving more toward primary care, more and more advanced practice nurses (APNs) are providing primary care to people with HIV. Because almost half of new cases of acquired immune deficiency syndrome continue to have homosexual or bisexual contact as a risk factor, the APN must be prepared to address the processes that are specific to the psychosocial development of homosexual persons. The most universal of these is coming out. Coming out is a stress-inducing process that can be magnified when combined with HIV infection. Moreover, with growing recognition of the presence and needs of gay and lesbian clients in the health care system, this challenge of helping people cope with the transitions of coming out extends beyond HIV care, from adolescence through old age. PMID- 8705928 TI - A review of the research section. PMID- 8705929 TI - Research utilization: once-a-day temperatures for afebrile patients. AB - CNSs AND OTHER Master's-prepared nurses are usually expected to lead research utilization (RU) efforts in organizations, but they often feel unprepared to do so. In this article, the need for clinical RU is discussed and ideas for implementation, using an adaptation of the Conduct and Utilization of Research in Nursing model, are described. The RU process is applied to research about once-a day temperatures in afebrile patients. To assist nurses who are implementing research-based practice, we list studies that were used for the practice change, summarize the research base on the topic, and offer an example of clinical use of practice guidelines. PMID- 8705930 TI - Recruitment of women into research studies: a nursing perspective. AB - RESEARCH TRIALS HAVE historically underrepresented women, especially women of ethnic minority and lower socioeconomic groups. Although these populations of women are disproportionately afflicted with human immunodeficiency virus (HIV), HIV studies have followed this pattern of underrepresentation. Nurses working in HIV research have developed a women-centered recruitment strategy that has been successful in enrolling women in clinical trials and epidemiological studies. Reasons for women's lack of inclusion and components of a strategy to remedy the situation are discussed. PMID- 8705931 TI - You really oughta wanna. PMID- 8705932 TI - Risk taking by health care workers. AB - HEALTH CARE WORKERS (HCWs) are at risk for acquiring infections such as human immunodeficiency virus, hepatitis B virus, hepatitis C, and tuberculosis from blood and body fluids. The Centers for Disease Control established guidelines known as universal precautions (UPs) to prevent transmission of blood-borne pathogens in hospital settings, but UPs are not universally followed. CNSs are in a unique position to enhance the practice of UPs and decrease the HCW's risk taking behavior. CNSs need to identify risk-taking behaviors and help HCWs make the right decision in risky situations. Reasons cited for noncompliance with UPs were habit, forgetfulness, influence of the nurse manager, and perceptions that barrier precautions hinder the ability to perform procedures successfully. Suggestions for improving compliance with UPs were better enforcement of guidelines, a policy mandating compliance, easy and available access to supplies, replacement of hazardous devices with safer ones, and discussion with HCWs on decision making in risky situations. CNSs possess the credibility in the clinical arena to effect change through clinical expertise, role modeling, and collaboration with HCWs. PMID- 8705933 TI - The legacy of Helena Willis Render (1896-1970). PMID- 8705934 TI - Is CNS flexibility in a cost-conscious world a liability? PMID- 8705935 TI - CNS role: an experience in retitling and role clarification. AB - A LITERATURE REVIEW and survey were undertaken to identify the appropriate titling for individuals in clinical nursing and other advanced practice roles, to establish components in each role, and to determine the educational level necessary for each role. Based upon review and survey results, a CNS task force at the University of Iowa Hospitals and Clinics proposed a change in the titles of nurses in advanced practice at their institution to reflect the American Nurses' Association's definition of advanced practice nursing and recommended graduate level education for these advanced positions. Five specific titles and primary role components were identified: advanced registered nurse practitioners. CNS, education nurse specialist, research nurse specialist, and informatics nurse specialist. Retitling aided in clearly identifying major responsibilities of the position, clarifying role expectations, and creating evaluation tools. PMID- 8705936 TI - CNS role evolution. AB - THE CNS ROLE has been actualized in a variety of ways. Flexibility-inherent in the role-and the revolution in health care consciousness tend to place the CNS at risk for criticism regarding value to the organization. At Vanderbilt University Medical Center, a CNS task force evaluated the current reality of CNS practice and recommended role changes to include the financial analysis of patient care. After incorporating a financial perspective into our present practice, we have embarked on an interesting journey of post-Master's degree study, that of the tertiary care nurse practitioner. This practice option could elevated the clinical and financial aspects of providing cost-effective health care to a more autonomous role form; however, the transition has been challenging. Since 1990, the American Nurses Association has recommended that nursing school curricula change to meet the needs of the health care environment and provide increased career flexibility through creating one advanced degree incorporating both CNS and NP functions. Swiftly moving past differences and toward similarities will bridge the gap for advanced practice nurses in the future. PMID- 8705937 TI - Medical family therapy: Therapeutic and financial benefits of family-centered care in the hospital. PMID- 8705938 TI - The psychiatric consultation-liaison nurse and medical family therapy. PMID- 8705939 TI - Facilitate change--improve care. PMID- 8705940 TI - Nurses' assessment and documentation of peripheral edema. AB - THE PURPOSE OF this study was to determine methods used by nurses to assess and document peripheral edema. The sample of 211 nurses at a large urban academic health center completed a survey regarding peripheral edema. Results indicated that nurses used a variety of methods to assess edema and documented their assessments on a number of different forms. Further, many stated that they were not proficient in assessment of edema and expressed the need for clear parameters to guide their practice. PMID- 8705941 TI - Unit discharge planning model. AB - THE UNIT DISCHARGE coordination model was implemented in January 1993 at the Children's Seashore House in Philadelphia, Pennsylvania, a 77-bed inpatient pediatric specialized care and rehabilitation institution. The model, which identifies staff nurses as primary discharge coordinators, has shown remarkable outcomes, evidenced by decreased lengths of stay, improved staff development, and better coordination of discharge resources. CNSs and nursing administrators will find this model cost effective, economically sound, and quality focused to enhance professional development at the staff level. PMID- 8705942 TI - When is the case manager role best filled by an advanced practice nurse? PMID- 8705943 TI - Nursing case management: relationships as a strategy to improve care. AB - MULTIPLE TRANSFERS, MULTIPLE caregivers, and an unpredictable hospital course may result in ineffective communication among patients, families, and the healthcare team, and a fragmented plan of care for complex patients. To address these concerns, CNSs in a tertiary hospital developed a nursing case management model for patients in a medical-surgical-neurological intensive care unit. Long-term relationships between nursing case managers (NCMs), patients, and families grounded the model. The NCM crossed traditional unit boundaries with the patient, improving communication among patient, family, and the healthcare team. Evaluation of the NCM's experience suggested four types of interventions: (a) telling the story, (b) advancing the plan of care, (c) maintaining values and beliefs, and (d) assisting with options and decisions. Case studies illustrate these interventions and demonstrate cost savings. PMID- 8705944 TI - Expanding collaborative CNS efforts for research utilization. PMID- 8705945 TI - The dependency of CNS practice. PMID- 8705946 TI - The psychogeriatric nurse in home health care: use of research to develop the role. AB - IN THIS ARTICLE, how one home health agency developed services for a high-risk group of clients based on a research study conducted on-site is described. Part of that study compared the cost of home care for cognitively impaired and cognitively intact persons over the age of 65. Although care for older adults with cognitive impairment was less expensive in terms of actual dollars, hospital and agency readmissions were significantly greater for those who were cognitively impaired, which led to greater total costs. Study results document that cognitively impaired older adults have special home care needs. Results were used to develop a special psychogeriatric program within the home health agency where data were collected. A CNS is now the key feature of this program; the CNS role in addressing the special needs of cognitively impaired home health clients is outlined. PMID- 8705947 TI - Chaos and self-organization as a consequence of spiritual disequilibrium. PMID- 8705948 TI - Nursing specialization in New South Wales, Australia. AB - SPECIALIZATION IN NURSING is a phenomenon that occurs worldwide. In New South Wales, Australia, the positions of clinical nurse specialist (CNS) and clinical nurse consultant (CNC) were formally established only in the last decade. How these positions developed and the responses to the roles are considered in this article. In 1990/1991, we surveyed 568 specialist nurses (CNSs and CNCs) working in the Sydney metropolitan area to identify their perceptions of what they ought to be doing and what they were actually doing in the performance of their roles. The results revealed discrepancies in perceptions of the potential role and the actual role. Comments indicated the challenging nature of the roles and thematic analysis of the comments identified problem areas related to role performance, such as workload and role conflict. Further research should be undertaken to explore the origins of role expectations and current role satisfaction. PMID- 8705949 TI - Development of clinical researchers. PMID- 8705950 TI - Improving self-efficacy in nursing research. AB - THE PURPOSE OF this project was to increase research activities of practicing nurses at one large healthcare facility. The members of the nursing research committee at this institution planned a participatory learning experience with the goal of increasing the nurses' self-efficacy toward conducting research. Increased self-efficacy was the impetus for stimulating increased research activities by the nurses. Measurable outcomes of this project included a significant increase in the participants' self-efficacy and implementation of a number of research projects. The Delphi technique proved useful in identifying nursing research priorities as well as in stimulating nurses to initiate and conduct clinical research. Project implementation and evaluation are described to help practicing nurses and their employers in pursuit of the same goal. PMID- 8705951 TI - No more educators? PMID- 8705952 TI - Nursing orientation: a continuous quality improvement story. AB - THIS ARTICLE ADDRESSES an orientation process utilizing a collaborative, multidisciplinary approach and principles of continuous quality improvement (CQI). This process successfully transitioned 63 graduate nurses into three different practice specialties, decreased the vacancy rate, and eliminated the need for agency nurses at Georgia Baptist Medical Center in Atlanta, Georgia. CNSs, acting as educators, clinical experts, and consultants, played a key role in developing, implementing, and evaluating this comprehensive program. PMID- 8705953 TI - Collaboration: more than a buzz word of the '90s. PMID- 8705954 TI - Establishing a collaborative practice in a comprehensive breast clinic. AB - BREAST CANCER IN women is a major health concern due to the high prevalence of subtle changes in breast tissue that are difficult to diagnose. Consequently, women require information about benign and malignant breast changes. Nurse practitioners (NPs) can play an important role in helping women with breast concerns by incorporating clinical expertise with teaching and counseling skills. Establishing clinics in which information, screening, and diagnosis of breast cancer are readily available will help decrease women's anxiety and enhance their ability to act as self-advocates within the healthcare system. A comprehensive breast clinic was developed, incorporating NP and physicians in a collaborative model of care. Defining roles within the model was an ongoing process as each provider developed areas of expertise and interest. The NP role emerged as an integral part of the practice, increasing the efficiency, availability, and effectiveness of clinic services. PMID- 8705955 TI - ECG changes in pediatric patients on tricyclic antidepressants, desipramine, and imipramine. AB - OBJECTIVE: To determine if there is an altered pattern of cardiac electrical activity in children treated with tricyclic antidepressants, desipramine, or imipramine, which may predispose these patients to sudden death. METHODS: All patients in a child psychiatry practice from 1989 to 1993 in Calgary, Alberta, treated with desipramine or imipramine with both pre- and post-treatment electrocardiograms (ECGs) were included in the study (n = 21; ages 8 to 17 years). Thirty-six blinded post-treatment ECGs were analysed for interval measurement and compared to the pretreatment ECGs. RESULTS: Drug dosages ranged from 25 mg to 125 mg per day and treatment duration ranged from 1 to 49 months. Seven of 21 patients were concurrently receiving an antipsychotic medication (pericyazine). The maximal increase in PR, and QRS, and QT interval changes were 40 msec and 70 msec, respectively, with most patients demonstrating no significant changes in the ECG intervals. The QT interval was corrected for heart rate (QTc). No significant arrhythmias or tachycardias were observed. ECG interval changes were not related to drug dosage, age, treatment duration or plasma levels. CONCLUSIONS: No consistent pattern of ECG interval changes including the QTc interval was observed in children on desipramine and imipramine. PMID- 8705956 TI - Key features of maltreatment of the infirm elderly in home settings. AB - OBJECTIVE: To identify contributory factors of elder abuse by caregivers in home settings. METHOD: Using a reliable instrument, visiting nurses rated observations symptomatic of abuse and neglect found in their current caseload of elderly patients. Their observations were also classified as related or unrelated to wilfull maltreatment. RESULTS: Regression analysis produced the following significant findings: 1. signs of poor physical care were found to be predictors of physical abuse; 2. signs of psychosocial distress and exploitation were identified as predictors of emotional abuse; 3. defensiveness and irritability shown by caregivers and strained family relationships, in general, were also associated with abuse. CONCLUSION: It appears that emotional abuse is more prevalent than, and not a necessary precursor of, physical abuse; however, reliable signs of impending or actual abuse of the elderly can be found in a home during visitation. PMID- 8705957 TI - [Psychometric properties of the French version of the Modified Mini-Mental State (3MS) in elderly patients evaluated in geriatric psychiatry]. AB - OBJECTIVE: This research evaluates the psychometric properties of the French version of the Modified Mini-Mental State (3MS), an instrument for the screening of dementia, in a sample of patients seen in geriatric psychiatry. METHODS: Subjects (N = 94, 63 to 93 years of age) were outpatients of a service in geriatric psychiatry. In addition to the 3MS, a battery of tests evaluating the cognitive functioning and depressive symptomatology was administered. RESULTS: Test-retest reliability coefficient was 0.87 for a subsample of 35 subjects. An exploratory factor analysis revealed 4 factors accounting for 63.2% of the variance. Cut-off scores of 79 to 80 (with years of education > or = 8) and 71 to 72 (with years of education < or = 7) presented the best combination sensitivity/specificity, respectively 0.80 and 0.96. CONCLUSIONS: This study documents the psychometric properties of the French version of the 3MS for use in patients seen in geriatric psychiatry, in suggesting cut-off scores which take into account the level of education. This study underlines the influence of sociodemographic variables on test performance. PMID- 8705958 TI - Re: Support groups and psychiatry. PMID- 8705959 TI - Re: Support groups and psychiatry. PMID- 8705960 TI - Re: Endocrine responses to fenfluramine challenge in chronic fatigue syndrome. PMID- 8705961 TI - Does electroconvulsive therapy exacerbate the elevation of creatinine phosphokinase caused by neuroleptics. PMID- 8705962 TI - Loxapine: an enigma. PMID- 8705963 TI - Wilson's disease and mental disturbances. PMID- 8705964 TI - Depression in the medically ill. PMID- 8705965 TI - The prevalence of major depressive disorder and low self-esteem in medical inpatients. AB - OBJECTIVE: To determine the prevalence of major depressive disorder in acutely ill medical inpatients, and the relationship of this to low self-esteem. METHOD: A total of 186 patients were interviewed 6 or 7 days following admission to detect the presence of DSM-IV major depressive disorder (MDD). Patients were assessed using a new brief psychiatric interview, the Silverstone Concise Assessment for Depression (SCAD), which has previously been validated for use in the physically ill. The cognitive function of the patients was measured, using the Mini-Mental State Examination (MMSE), with patients scoring less than 22 on the MMSE being excluded from the study. The patients' self-esteem was also assessed, using the Rosenberg self-esteem rating scale. The severity and type of the patients' medical illness, and the recognition of psychiatric illness by both nurses and physicians were also noted. RESULTS: The results showed that 18 patients (9.7%) were depressed. The depressed patients were significantly younger than the nondepressed patients (mean age 46.3 +/- 3.9 years versus 57.1 +/- 1.5 years, respectively) and were significantly more likely to be female (61% versus 44%, respectively). The depressed patients had a significantly lower self-esteem than the nondepressed patients, whose self-esteem was no different from the general population. However, the depressed patients were not more severely ill than the nondepressed patients. The results also demonstrated that both nurses and physicians were poor at recognizing the presence of major depression, with nurses recognizing 33% of cases compared to 22% for medical staff. CONCLUSIONS: The results from this study demonstrate that while there is an increase in the incidence of depression in medically ill patients, this is not as great as has been previously reported, and is not related to severity of illness. The results from this study, therefore, are in keeping with other recent findings which show that the prevalence rates for MDD in medical patients is between 5% to 10% rather than the previously accepted range of 20% to 40%. PMID- 8705966 TI - Antisocial personality disorder: a biopsychosocial model. AB - OBJECTIVE: To propose an etiological model of antisocial personality disorder that is grounded in empirical data. METHOD: Recent research findings are reviewed that clarify our understanding of the etiology and course of antisocial pathology. RESULTS: Neither biological nor psychological factors fully account for the development of this disorder. Epidemiological studies show that there are strong cross-cultural differences in its prevalence, pointing to the importance of social factors in its etiology. Outcome research shows that antisocial personality only partially remits with time, and that most patients continue to be dysfunctional in later life. No treatment modality has been shown to be effective. CONCLUSIONS: All these empirical findings can be accounted for by a biopsychosocial model of antisocial personality disorder. PMID- 8705967 TI - Impulsivity, defensive functioning, and borderline personality disorder. AB - OBJECTIVE: To replicate previous research suggesting that impulsivity highly predicts current DIB(R) score and social functioning, with the additional inclusion of other measures, including defensive functioning. METHOD: Correlational analyses between impulsivity and other measures, and regression analyses with DIB(R) and SAS-SR as outcome measures, and impulsivity and other measures as predictor variables, were performed on data derived from n = 57 initially borderline personality disorder (BPD) subjects recruited as part of a 7 year follow-up study of BPD. RESULTS: Data showed strong correlations between the elements of impulsivity described previously and defensive functioning. The initial study results were repeated, and only a minor contribution from defensive functioning additionally contributed to the regression models. No other variables entered the model, unless anger was dropped from the variables entered into the analysis. CONCLUSIONS: The results may contribute to a better definition of the term "impulsivity" as related to BPD, and may lead to further, improved research into the cause, treatment, and prognosis of BPD. PMID- 8705968 TI - The assessment of competence to make a treatment decision: an empirical approach. AB - OBJECTIVE: To compare physicians' judgements of competency in routine clinical practice with the findings obtained from a structured clinical interview. METHODS: Ninety-six patients referred for electroconvulsive therapy were administered the Competency Interview Schedule (CIS) prior to their first treatment. Cluster analysis was employed to categorize patients to 1 of 5 cluster centres represented by case studies previously judged competent or incompetent by lawyers and health professionals. RESULTS: A match-mismatch table revealed 88% (N = 66) of the 75 patients found competent by the attending physician and 90.5% (N = 19) of the 21 patients found incompetent by the attending physician were classified in agreement with the CIS. The 9 misclassified patients found competent by the attending physician but classified incompetent by the CIS had consented to treatment. The 2 misclassified patients found incompetent by the attending physician but classified competent by the CIS had refused treatment. Examination of individual item scores from the CIS indicated that, in some cases, a different standard of competency was applied in routine clinical practice depending upon the patient's treatment decision. CONCLUSIONS: The CIS is presented as a useful guide for clinicians with an interest in competency evaluations but caution is advised in using the instrument to make formal evaluations of competency owing to the imprecise definition of competency in various jurisdictions. PMID- 8705969 TI - Intensive use of mental health care. AB - OBJECTIVE: To describe the profile of the intensive use of mental health services over a 4-year period in a population of 1.1 million people. METHODS: Data obtained from computerized hospital separation records and physician reimbursement claims were combined to form patient-based histories of mental health care utilization. Users of mental health services in a 24-month period were hierarchically classified as having a psychotic disorder (ICD-9-CM 295-299) or a nonpsychotic disorder (ICD-9-CM 300-301, 306-309, 311). Intensive use was defined as 12 or more contact months or a minimum of 2 episodes of therapy in the 24-month period. The cohort of intensive users were followed over the subsequent 24-month interval to describe the persistence of intensive use. RESULTS: In the initial observation periods, intensive users constituted 27.4% of individuals in treatment for psychotic disorder and 4.4% of persons in treatment for nonpsychotic disorder. These 2 groups, which represent 7.4% of all users of mental health care, were responsible for 53% of physician services, 72.7% of contacts with psychiatrists, and 64.4% of acute psychiatric bed days in the initial period. In the follow-up period, intensive use status was replicated by 44.6% of the cohort. CONCLUSIONS: The diagnostic and therapeutic characteristics of intensive users of mental health services are heterogeneous. There is substantial persistence of intensive mental health service use over time. PMID- 8705970 TI - An ethicist's commentary on the veterinarian receiving complaints from swine barn workers concerning respiratory problems. PMID- 8705971 TI - The Canadian rural practitioner market. PMID- 8705972 TI - Trends in bovine abortions submitted to the Ontario Ministry of Agriculture, Food and Rural Affairs, 1993-1995. PMID- 8705973 TI - Serum-free thyroxine concentrations, measured by chemiluminescence assay before and after thyrotropin administration in healthy dogs, hypothyroid dogs, and euthyroid dogs with dermathopathies. AB - The purpose of this study was to determine the usefulness of free thyroxine (FT4) measured by chemiluminescence in evaluating thyroid function in dogs. Total thyroxine (TT4) concentration measured by radioimmunoassay (RIA) and FT4 measured by chemiluminescence were evaluated in 30 healthy dogs, 60 euthyroid dogs with concurrent dermatopathies, and 30 hypothyroid dogs before and after intravenous stimulation with 1 or 2 IU of thyrotropin (TSH). Median basal TT4 and median TT4 concentrations at 4 h post-TSH administration were not significantly different (P < 0.0001) between healthy dogs and euthyroid dogs with dermatopathies, but were significantly higher than those in hypothyroid dogs. In healthy dogs, the median TT4 concentrations at 4 and 6 h post-TSH administration were not significantly different. Median basal FT4 and median FT4 concentrations at 4 h post-TSH administration in healthy dogs were significantly lower (P < 0.0001) than those in euthyroid dogs with dermatopathies, but significantly higher than the same parameters in hypothyroid dogs. There was a significant difference between the median FT4 concentrations at 4 h post-TSH administration and median basal FT4 concentrations for healthy dogs and euthyroid dogs with dermatopathies, but not for hypothyroid dogs. Lastly, in healthy dogs, median FT4 concentrations at 4 and 6 h post-TSH administration were not significantly different. Free thyroxine measured by chemiluminescence was highly correlated (P < 0.0001; Spearman r = 0.91) with FT4 measured by the reference method for free hormone analysis, namely, equilibrium dialysis, when sera from 56 dogs were used. PMID- 8705974 TI - Comparison of herniorrhaphy versus clamping of umbilical hernias in horses: a retrospective study of 93 cases (1982-1994). AB - Many uncomplicated umbilical hernias have been managed successfully in foals by the application of a hernia clamp. Isolated reports of complications following clamp application have led some authors to suggest that it is an unsuitable method of treatment. Little information has been published comparing the complication rates associated with the use of hernia clamps and herniorrhaphy in the treatment of umbilical hernias. The purpose of this retrospective study was to report the characteristics of clinical cases of umbilical hernia and to compare the complication rates following these 2 treatment approaches. Information was collected from records or from owners to identify the occurrence of complications and owner satisfaction following treatment. Of 93 cases, 10 complicated and 18 uncomplicated hernias were treated by herniorrhaphy, 40 uncomplicated hernias were treated by clamping, 1 originally uncomplicated hernia was treated by both techniques, and 24 cases were untreated. Nineteen percent of uncomplicated hernias treated by herniorrhaphy, and 19% of those clamped developed minor complications. This study demonstrates that although minor complications may be associated with either technique, they generally do not result in significant morbidity. PMID- 8705975 TI - A field trial to evaluate a Mycoplasma hyopneumoniae vaccine: effects on lung lesions and growth rates in swine. AB - A killed Mycoplasma hyopneumoniae vaccine was evaluated in a single swine herd in which the farrowing barn and weaner rooms were on one Mycoplasma-free farm, while the growing and finishing barn was on a separate farm on which Mycoplasma was present. The study was carried out in a cohort of pigs born in a 12-week period. Pigs born in 6 of the 12 wk were vaccinated and the rest were left as controls. The vaccine was administered twice at approximately 3 and 6 wk of age. Carcass characteristics, lung lesions, and growth rates were recorded on 893, 390, and 220 pigs, respectively. The vaccine reduced the prevalence of pneumonic lesions in slaughter hogs from 69% to 36% (P < 0.001). It also appeared to reduce the prevalence of pleuritis from 20% to 13%, but the difference was only statistically significant at P = 0.07. The vaccine had no effects on carcass characteristics except that carcasses of vaccinated pigs were, on average, 1 kg heavier than those of nonvaccinated pigs, and a smaller percentage of vaccinated pigs were shipped "light" (carcass weight < 70 kg). Two methods were used to estimate the effect of the vaccine on growth rates (as measured by days to 80 kg carcass weight) resulting in estimates of 11 and 2 d reduction attributable to vaccination, respectively. The latter estimate was probably an underestimate for reasons discussed in the paper. PMID- 8705976 TI - Generalized encephalitozoonosis in a Jersey wooly rabbit. PMID- 8705977 TI - Acute canine polyradiculoneuritis. PMID- 8705978 TI - Treatment of mandibular and dental fractures in a young dog. PMID- 8705979 TI - Critical thinking: how the characteristic coronoid lesion evolves. PMID- 8705980 TI - "Lien on me"--veterinary liens for accounts receivable. PMID- 8705981 TI - [Cancer of the larynx]. AB - Cancer of the larynx and total laryngectomies have devastating consequences on the patient and his/her family. During a specialized field placement in family nursing care, the author used the Calgary Family Assessment Model to better understand the relationship between the disease, the surgical procedure and the resulting family dynamics. She found that nursing interventions not only helped the family regain a sense of balance but facilitated the patient's adjustment process. PMID- 8705982 TI - Family systems nursing education. PMID- 8705983 TI - Tuning in new parents. PMID- 8705984 TI - Candida: the hidden deterrent to breastfeeding. AB - The '80s and '90s have been characterized by an increase in the number of breastfeeding mothers. The added health benefits of breastfeeding for both mother and child, the father's influence, maternal age, family practices and education all may contribute to a mother's decision to breastfeed. Although the incidence of mothers choosing to breastfeed their infants has increased, both research and practice indicate that a significant number of mothers stop breastfeeding long before their target date. Among the many reasons given for the early cessation of breastfeeding is the complaint of persistently sore and red nipples. Such a complaint should raise a red flag to the hospital or community nurse. The pain and redness could be due to Candida albicans. PMID- 8705985 TI - Embracing the family. AB - Family-focused care has long been valued by the profession of nursing, but until recently it has not received the recognition it deserves in academic and health care settings. Now, with the downsizing of institutions, higher in-patient acuity with resultant family stress, and the move to providing care and treatment in the patients' homes or communities, nurses are increasingly required to be knowledgeable and skilled in communicating and problem solving with families. At the same time, nurses are also being asked to provide expedient, qualitative care and treatment that is research theory-based, cost-effective and produces positive outcomes. These expectations come at a turning point in Canada's health care history when there are shrinking resources in health care institutions, and a concomitant demand for improved consumer satisfaction. PMID- 8705986 TI - Windows on the family. AB - Events in the last few decades have resulted in major changes in the structure of families and in the roles and relationships of family members. As nurses, we deal with the stress-related problems resulting from these changes, and as family roles change, so too do our nursing actions. But exactly what these changes are and how they have affected the family vary according to the perspective of the viewer. PMID- 8705987 TI - [The life of the couple and feelings of parental competence]. AB - With the birth of their first child a couple must learn to adapt to their new role as parents. During this learning process, their degree of comfort with their new identities as mother and father is linked to how they develop as competent parents. According to the ecological approach to human development, several variables influence parental competency. The marital relationship and the family dynamics are of special interest to the nurse who often has to evaluate the family system and intervene during the transition period to parenthood. This descriptive-correlative study is in direct response to provincial priorities aimed at promoting parenthood and providing better support to parents. Surveys were mailed to 133 couples with a one-year-old child to determine the nature of the link between the marital relationship, the family dynamics and the feeling of parental competency. The results confirm the importance of the nurses involved in evaluating these variables so that effective interventions can be developed to support and assist the couple--thus reinforcing their feelings of parental competency. PMID- 8705988 TI - New moms and self medication. AB - "Is there any reason why healthy women should not have access to simple pain medication when they require it?" This is the question pharmacists at the Salvation Army Grace Hospital in Ottawa were asked in addressing the needs of new mothers. And, while post-partum self-medication programs have existed for over a decade in Canada, the Grace has developed and implemented a comprehensive, patient-focused, multidisciplinary system with an innovative and user-friendly approach that benefits patients and staff. PMID- 8705989 TI - Taryn's story. AB - Hospitalization of a young child with a chronic illness is a nightmare for the child and family. And the nightmare can get worse for children with multiple health care problems managed by several sub-specialty teams. The child feels anxious; parents don't know who to turn to for information and support. Collaborative nursing and family interventions can help relieve some of these problems. PMID- 8705990 TI - Balancing family, work and school. PMID- 8705991 TI - Children and consent to treatment. PMID- 8705992 TI - Is privatization the answer? AB - Concern is being expressed by some that Canada's publicly funded health care system can no longer be maintained and that a privately funded system will help ease the pressure on our public system. Indeed, the issue of privatizing Canada's health care system promises to be one of this fall's hottest debates. PMID- 8705993 TI - Nuclear signaling induced by ionizing radiation involves colocalization of the activated p56/p53lyn tyrosine kinase with p34cdc2. AB - The Src-like protein-tyrosine kinase p56/p53lyn associates with cell membranes and transduces signals from activated cell surface receptors. In the present work, cell fractionation and confocal microscopy studies demonstrate expression of Lyn in the nucleus. We also demonstrate that exposure of intact cells to ionizing radiation is associated with selective activation of nuclear Lyn. Similar findings have been obtained following irradiation of purified nuclei. Immunoprecipitation studies of nuclear lysates demonstrate radiation-induced binding of Lyn to p34cdc2. Nuclear colocalization of Lyn with Cdc2 has been confirmed by confocal microscopy. Other studies with glutathione S-transferase Lyn fusion proteins demonstrate that the binding of Lyn to nuclear Cdc2 is associated with inhibition of Cdc2 activity. These findings suggest that the association of activated Lyn with Cdc2 in the nucleus may contribute to regulation of a DNA damage-dependent premitotic checkpoint. PMID- 8705994 TI - Somatic and germline mutations of the BRCA2 gene in sporadic ovarian cancer. AB - The breast and ovarian cancer susceptibility gene BRCA2 has recently been isolated. A role for BRCA2 in sporadic breast and ovarian cancer has been suggested by loss of heterozygosity (LOH) studies which show frequent LOH in the BRCA2 region at chromosome 13q12. In addition, the observation of nonrandom loss of the wild-type chromosome in a breast/ovarian cancer family which shows linkage to BRCA2 suggests it may act as a tumor suppressor gene. To determine the extent of somatic alteration involving BRCA2 in sporadic ovarian cancer, 50 tumors were analyzed for mutations throughout the entire BRCA2 coding region. Mutations predicted to result in truncation of the BRCA2 protein were detected in four tumors. Analysis of germline DNA revealed two of these alterations to be of somatic origin. In addition, all four tumors exhibited loss of the second BRCA2 allele as predicted by Knudson's hypothesis for a tumor suppressor gene. These results suggest that, as is the case with BRCA1, somatic mutations of BRCA2 are infrequent in sporadic ovarian cancer, despite the relatively high frequency of LOH detected around the BRCA2 locus. PMID- 8705995 TI - Hypersensitizing effect of pluronic L61 on cytotoxic activity, transport, and subcellular distribution of doxorubicin in multiple drug-resistant cells. AB - The present study demonstrated that poly(oxypropylene) and poly(oxyethylene) block copolymer pluronic L61 (L61)-hypersensitized multidrug-resistant CHRC5 Chinese hamster ovary cells and MCF-7/ADR human breast carcinoma cells to the cytotoxic action of doxorubicin (Dox). CHRC5 and MCF-7/ADR cells manifested 290- and 700-fold increases, respectively, in their sensitivity to Dox/L61 formulation compared with free Dox. Their sensitive counterparts Aux-B1 and MCF-7 displayed only marginal or no increase at all in their response to Dox/L61. The study of the drug transport performed by flow cytometry showed that L61 enhanced the drug uptake and reduced the P-glycoprotein-mediated drug efflux. Visualization of Dox subcellular distribution in CHRC5 cells by fluorescent microscopy revealed that Dox was sequestered in cytoplasmic vesicles, whereas incubation of the cells with Dox/L61 altered the drug compartmentalization by releasing the drug from these vesicles and shifting it to the nucleus. These findings suggested that the hypersensitive response of multidrug-resistant cells to the action of Dox/L61 was caused by an increase in the drug accumulation and changes in its subcellular distribution. PMID- 8705996 TI - High frequency of p16 (CDKN2/MTS-1/INK4A) inactivation in head and neck squamous cell carcinoma. AB - The tumor suppressor gene p16 (CDKN2/MTS-1/INK4A) can be inactivated by multiple genetic mechanisms. We analyzed 29 invasive primary head and neck squamous cell carcinomas (HNSCC) for p16 inactivation with immunohistochemistry utilizing a new monoclonal antibody (mAb), DCS-50. p16 staining of the primary lesions was correlated with genetic analysis including: (a) detailed microsatellite analysis of markers at the p16 locus to detect homozygous deletion; (b) sequence analysis of p16; and (c) Southern blot analysis to determine the methylation status of the 5' CpG island of p16. Twenty-four of 29 (83%) head and neck squamous cell carcinoma tumors displayed an absence of p16 nuclear staining using immunohistochemistry. Of these 24 tumors, we found that 16 (67%) harbored homozygous deletions, 5 (21%) were methylated, 1 displayed a rearrangement at the p16 locus, and 1 displayed a frameshift mutation in exon 1. These data suggest that: (a) inactivation of the p16 tumor suppressor gene is a frequent event in squamous cell carcinomas of the head and neck; (b) p16 is inactivated by several distinct and exclusive events including homozygous deletion, point mutation, and promoter methylation; and (c) immunohistochemical analysis for expression of the p16 gene product is an accurate and relatively simple method for evaluating p16 gene inactivation. PMID- 8705997 TI - Two differentially expressed genes in normal human prostate tissue and in carcinoma. AB - Alterations in transcriptional control may contribute directly to carcinogenesis. Using the differential display technique in prostate cancer cells compared to normal prostate epithelial cells, we identified a down-regulated gene and an up regulated gene in cancer cells. The down-regulated gene encodes human epithelial tropomyosin (TMe1), a member of the family of actin filament-binding proteins. The up-regulated gene encodes cytochrome c oxidase subunit VIc (COSVIc), a protein of the respiration chain in the mitochondrial inner membrane. The differential display pattern was confirmed by Northern hybridization in both prostate tissue and cell lines. In situ hybridization of malignant prostate epithelial tissue using a digoxigenin-labeled antisense riboprobe detected strong staining for mRNA of COSVIc, as opposed to very weak staining in normal prostate epithelium. The expression pattern of COSVIc may be a useful marker for studying the alteration of energy metabolism in cancer cells and for the diagnosis of prostate cancer. PMID- 8705998 TI - In vitro induction of benzo(a)pyrene diol epoxide-DNA adducts in peripheral lymphocytes as a susceptibility marker for human lung cancer. AB - Given the same exposure, DNA adduct profiles can be considered as a phenotypic marker for carcinogen metabolism and DNA repair, which may reflect individual susceptibility to chemical carcinogenesis. Based on this notion, we have established a straightforward assay that measures induced DNA adducts in peripheral lymphocytes exposed in vitro to a model carcinogen, benzo(a)pyrene diol epoxide (BPDE) by 32P-postlabeling. To test the hypothesis that the levels of induced DNA adducts are a predictor for cancer risk, we conducted a pilot study of 21 lung cancer patients and 41 healthy frequency-matched controls. We found that the peripheral lymphocytes of cancer patients tended to accumulate higher levels of BPDE-DNA adducts than controls did (mean +/- SE of relative adduct labeling x 10(7) value; 59.6 +/- 12.0 versus 39.4 +/- 6.1 for cases and controls, respectively; P = 0.09). Using the tertile relative adduct labeling value of controls (10 adducts/10(7) nucleotides) as the cutoff point, 18 of 21 cases and 23 of 41 controls distributed above this level (odds ratio, 4.7; 95% confidence interval, 1.2-18.5). In logistic regression analysis, the level of induced adduct was an independent risk factor (odds ratio, 6.4; 95% confidence interval, 1.3-29.4) after adjustment for the potential confounding factors, i.e., age, sex, ethnicity, and smoking. Stratified analyses showed that greater differences in DNA adduct levels induced by BPDE between cases and controls were observed in individuals younger than 65 years and in nonsmokers. Despite the small sample size, the significant association between the level of BPDE-induced DNA adducts and risk for lung cancer suggests that this assay is a promising method for further investigations. PMID- 8705999 TI - Frequent coordinated overexpression of the MRP/GS-X pump and gamma glutamylcysteine synthetase genes in human colorectal cancers. AB - We have recently shown that multidrug resistance-associated protein (MRP) and gamma-glutamylcysteine synthetase (gamma-GCS) heavy subunit genes are coordinately overexpressed in cisplatin-resistant human leukemia cells (T. Ishikawa et al. J. Biol. Chem., 271: 14981-14988, 1996). Using the RNase protection assay, we examined expression levels of these genes in colon tumor and nontumorous biopsy specimens from 32 cancer patients who had not been treated with chemotherapy. Increased mRNA levels (P < 0.001) of MRP and gamma-GCS genes were observed in 16 (50%) and 20 (62%) tumor samples, respectively. More importantly, all of the 16 (100%) MRP-overexpressing tumor specimens also exhibited higher levels of gamma-GCS mRNA than those in the matched nontumorous specimens. The correlation coefficient between MRP and gamma-GCS mRNA levels was r = 0.78 for all of the tumor samples studied. These results strongly suggest that MRP and gamma-GCS genes are coordinately up-regulated during colorectal carcinogenesis. PMID- 8706000 TI - Transforming growth factor beta 1 suppresses genomic instability independent of a G1 arrest, p53, and Rb. AB - Alterations in expression of or responsiveness to transforming growth factor beta (TGF-beta) are frequently found in human and animal epithelial cancers and are though to be important for loss of growth control in the neoplastic cell. We show here that keratinocyte cell lines from mice with a targeted deletion of the TGF beta 1 gene have significantly increased frequencies of gene amplification in response to the drug N-phosphonoacetyl-L-aspartate (PALA) compared to TGF-beta 1 expressing control keratinocyte cell lines. In contrast to the control lines, the PALA-mediated G1 arrest did not occur in the TGF-beta 1 null keratinocytes despite the presence of wild-type p53 in both genotypes. Exogenous TGF-beta 1 suppresses gene amplification in the null keratinocytes at concentrations that do not cause a G1 growth arrest and in human tumor cell lines that are insensitive to TGF-beta 1-mediated growth inhibition. The pathway of TGF-beta 1 suppression is independent of the p53 and Rb genes, but requires an intact TGF-beta type II receptor. These studies reveal a novel TGF-beta-mediated pathway regulating genomic stability and suggest that defects in TGF-beta signaling may have profound effects on tumor progression independent of cell proliferation. PMID- 8706001 TI - Pregnancy-dependent growth of mammary tumors is associated with overexpression of insulin-like growth factor II. AB - We demonstrate that although IGF-II gene expression is approximately 3-fold higher in 9,10-dimethyl-1,2-benzanthracine (DMBA)-induced rat mammary tumors (MTs) than in nonneoplastic breast tissue, IGF-II mRNA abundance in DMBA-induced MTs is approximately 130-fold higher in pregnant as compared to nonpregnant hosts. This correlated with accelerated tumor growth in pregnant hosts. Immunohistochemical studies of DMBA-induced MTs with an anti-IGF-II antibody showed an intense staining of tumor cells for IGF-II, whereas a very low staining signal was observed for normal epithelial cells in the lobules. A similar immunostaining pattern was observed in three of three human ductal cancers and adjacent normal breast tissue obtained during pregnancy. DMBA-induced MTs expressed high levels of type I receptor for IGFs as determined by Northern blots. In vitro studies confirmed that IGF-II is a mitogen for neoplastic epithelial cells derived from DMBA-induced MTs. These results demonstrate that hormonal changes associated with pregnancy accelerate breast cancer cell proliferation in the DMBA-induced MT model and suggest that this acceleration is mediated by up-regulation of IGF-II expression within neoplasms. PMID- 8706002 TI - Methylation of the estrogen receptor CpG island in lung tumors is related to the specific type of carcinogen exposure. AB - Promoter methylation has recently been shown to be an alternative to mutation in inactivating tumor suppressor genes in human neoplasia. Although specific carcinogen exposures have been associated with characteristic mutation patterns in genes, the factors that lead to promoter hypermethylation remain unknown. One gene target for inactivation through promoter methylation is the estrogen receptor (ER). The purpose of this investigation was to determine the methylation status of this gene in lung tumors from smokers and those who never smoked and in rodents exposed to specific environmental carcinogens. Promoter methylation at the ER locus was detected in 4 of 11 tumors from never-smokers (36.4%) and 7 of 35 tumors from smokers (20%, P < 0.001). Lung tumors induced by the tobacco derived carcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1- butanone also had a low incidence (16.7%) of ER methylation. In marked contrast, spontaneous and plutonium-induced tumors had a very high (81.8%) incidence of ER methylation. X ray-induced tumors had an intermediate frequency of ER methylation (38.1%). The presence of ER methylation was associated with absent ER expression in rodent lung cancer cell lines. These results show for the first time that gene-specific promoter methylation can be modulated differentially depending on carcinogen exposure. PMID- 8706003 TI - Separate pathways for p53 induction by ionizing radiation and N-(phosphonoacetyl) L-aspartate. AB - The tumor suppressor gene product, p53, appears to be a significant participant in signaling pathways that mediate cellular responses to cytotoxic stresses. In particular, p53 appears to be a critical determinant of whether the cell lives or dies and how it progresses through the cell cycle after the cytotoxic exposure. Many of the molecular details for these signaling pathways remain to be elucidated, and whether all cytotoxic signals utilize the same pathway to increase p53 expression is not clear. Here, we demonstrate the existence of cell types in which the induction of p53 and associated G1 arrest by the antimetabolite, N-(phosphonoacetyl)-L-aspartate (PALA), is defective, whereas p53 induction and G1 arrest induced by ionizing radiation are intact. These observations demonstrate the existence of genetic defects that can alter p53 induction and associated cellular outcomes after some, but not all, cytotoxic insults and suggest distinct pathways of p53 induction by PALA and ionizing radiation. PMID- 8706004 TI - High prevalence of the 999del5 mutation in icelandic breast and ovarian cancer patients. AB - Studies on Icelandic breast cancer families have shown that most of them segregate a 999del5 BRCA2 mutation. Here, we report the frequency of the 999del5 BRCA2 mutation in an Icelandic control population and four different groups of cancer patients diagnosed with (a) breast cancer; (b) ovarian cancer; (c) prostate cancer (patients younger than 65 years); and (d) other cancer types. The proportions of individuals carrying the mutation were 0.4% in the control population and in the patient groups 8.5%, 7.9%, 2.7%, and 1.0%, respectively. Our results indicate that BRCA2 confers a very high risk of breast cancer and is responsible for a substantial fraction of breast and ovarian cancer in Iceland, but only a small proportion of other cancers. PMID- 8706005 TI - Anti-epidermal growth factor receptor monoclonal antibody 225 up-regulates p27KIP1 and induces G1 arrest in prostatic cancer cell line DU145. AB - Autocrine production of transforming growth factor alpha and overexpression of the epidermal growth factor receptor (EGFR) may contribute to androgen independent prostatic cancer growth at both primary and metastatic sites. Previously, we showed that human EGFR-blocking monoclonal antibody mAb225 inhibited the growth of DU145 human prostatic cancer cells. Here we explore the hypothesis that mAb225 may act by interfering with cell cycle traversal in these cells. Treatment with mAb225 induced G1 arrest, which was accompanied by a marked decrease in CDK2-, cyclin A-, and cyclin E-associated histone H1 kinase activities, and a sustained increase in cell cycle inhibitor p27KIP1. The increased p27KIP1 levels were attributable to elevation of both transcription and translation. CDK2 associated with p27KIP1 was increased in mAb225-treated DU145 cells. The retinoblastoma-related protein p130 remained hypophosphorylated in these retinoblastoma-negative cells. These studies demonstrate that the antiproliferative effect of EGFR blockade in DU145 cells may be mediated by up regulation of p27KIP1 at both the mRNA and protein levels. PMID- 8706006 TI - Vitamin B6 and cancer: synthesis and occurrence of adenosine-N6-diethylthioether N-pyridoximine-5'-phosphate, a circulating human tumor marker. AB - In the course of studies aimed at deciphering the metabolic transformations of [3,4-14C] and [3H]C6-pyridoxine hydrochloride by tumor-bearing rats and tumor cells in culture, biosynthesis of a novel labeled product was observed. Its production began with the onset of tumor growth and increased as cell proliferation increased. Chemical, enzymatic, precursor labeling, and analytical tests on the isolated product indicated this product as adenosine-N6 diethylthioether-N-pyridoximine-5'-phosphate (compound 1). In confirmation, the chemical synthesis and characterization of compound 1 are presented in this study. In addition, blood samples from 28 normal subjects, 28 cancer patients with different malignancies, and 39 patients with a variety of other than cancer ailments were screened for compound 1 on a blind basis using reverse phase ion paired high-performance liquid chromatography. The results show that the level of the vitamin B6 conjugate in the circulation of control subjects, cancer patients in remission, and patients with other diseases was only minimal. Cancer patients with active disease had 3-4-fold higher levels (P < 0.00001). Our results also confirm previous findings regarding the structure of compound 1 and show its potential value as a circulating human tumor marker that could be successfully used for cancer detection. PMID- 8706007 TI - Busulfan-glutathione conjugation catalyzed by human liver cytosolic glutathione S transferases. AB - We have examined the catalytic activity of glutathione S-transferases (GST) in the conjugation of busulfan with glutathione (GSH) in human liver cytosol, purified human liver GST, and cDNA-expressed GST-alpha 1-1. Human liver microsomes and cytosol were incubated with 40 microM busulfan and 1 mM GSH. Cytosol catalyzed the formation of the GSH-busulfan tetrahydrothiophenium ion (THT+) in a concentration-dependent manner, whereas microsomes lacked activity. The total and spontaneous rates of THT+ formation increased with pH (pH range, 6.50-7.75), with the maximum difference at pH 7.4. Due to the limited aqueous solubility of busulfan, a K(m) for busulfan was not determined. The intrinsic clearance (Vmax/K(m)) of busulfan conjugation was 0.167 microliter/min/mg with 50 1200 microM busulfan and 1 mM GSH. GSH Vmax and K(m) for busulfan conjugation were 30.6 pmol/min/mg and 312 microM, respectively. Ethacrynic acid (0.03-15 microM) inhibited cytosolic busulfan-conjugating activity with 40 microM busulfan and 1 mM GSH. Enzyme-mediated THT+ formation was decreased 97% by 15 microM ethacrynic acid with no effect on the spontaneous reaction. In incubations with affinity-purified liver GST and GST-alpha 1-1, the intrinsic clearance for busulfan conjugation was 0.87 and 2.92 microliters/min/mg, respectively. Busulfan is a GST substrate with a high K(m) relative to concentrations achieved clinically (1-8 microM). PMID- 8706008 TI - Chaotic oscillations in cultured cells: rat prostate cancer. AB - Normal prostate epithelial cells exhibit uniformity of structure, function, and DNA content. This uniformity is dramatically perturbed in cancer with the development of variance associated with tumor cell heterogeneity. The development of this kind of diversity is paralleled in models of chaotic oscillators that produce multiple pseudosteady states. We have tested prostatic cancer cells in culture for the presence of chaos by comparing the micromotion of two related rat prostate cancer cell lines that exhibited large differences in motility and metastatic potential. In these extremes of cancer cell types, our data suggest that the three criteria which characterize a chaotic oscillation are fulfilled by their cellular micromotions: (a) absence of defined regularity in the time series as evidenced using Fourier analysis and visual inspection; (b) determinism as evidenced by attractor reconstruction; and (c) sensitive dependence on initial conditions as evidenced by a positive Lyapunov exponent. Cellular motion was studied by using an electronic cell impedance sensor which records, in real time, the fluctuations in the resistive and capacitive properties of cells cultured on recording electrodes. Our data and a preliminary screen of other cell types support a model of established cell lines in culture as chaotic oscillators. PMID- 8706009 TI - Identification and properties of a major plasma metabolite of irinotecan (CPT-11) isolated from the plasma of patients. AB - Irinotecan [7-ethyl-10-[4-(1-piperidino)-1-piperidino]carbonyloxycamptothecin (CPT-11)] is a promising water-soluble analogue of camptothecin [S. Sawada et al., Chem. & Pharm. Bull. (Tokyo), 39: 1446-1454, 1991]. We have reported previously the presence of an important polar metabolite, in addition to 7-ethyl 10-hydroxycamptothecin (SN-38) beta-glucuronide, in samples of plasma taken from patients undergoing treatment with CPT-11 (L.P. Rivory and J. Robert, Cancer Chemother. Pharmacol. 36: 176-179, 1995; L. P. Rivory and J. Robert, J. Cromatogr., 661: 133-141, 1994). Plasma samples (0.5 ml) containing comparatively large amounts of this metabolite were extracted by solid-phase columns and subjected to high-performance liquid chromatography and mass spectrometry in parallel to fluorometric detection. The metabolite yielded [M + 1] ions with a m/z of 619, representing the addition of 32 atomic mass units to CPT-11. Purified fractions were subjected to proton nuclear magnetic resonance, and the structure determined, 7-ethyl-10-[4-N-(5-aminopentanoic acid)-1 piperidino]carbonyloxycampothecin (APC), was further validated following synthesis. Like CPT-11, APC was found to be only a weak inhibitor of the cell growth of KB cells in culture (IC50, 2.1 versus 5.5 micrograms/ml for CPT-11 and 0.01 microgram/ml for SN-38, the active metabolite of CPT-11) and was a poor inducer of topoisomerase I DNA-cleavable complexes (100-fold less potent than SN 38). In contrast to CPT-11, APC was not hydrolyzed to SN-38 by human liver microsomes or purified human liver carboxylesterase. Furthermore, APC did not inhibit the hydrolysis of CPT-11 in these preparations. Interestingly, APC was only a weak inhibitor of acetylcholinesterase in comparison to CPT-11 and neostigmine. It appears likely, therefore, that APC does not contribute directly to the activity and toxicity profile of CPT-11 in vivo. PMID- 8706010 TI - Dose-dependent differences in the mutational profiles of (-)-(1R,2S,3S,4R)-3,4 dihydroxy-1, 2-epoxy-1,2,3,4-tetrahydrobenzo[c]phenanthrene and its less carcinogenic enantiomer. AB - Chinese hamster V-79 cells were treated with high cytotoxic or low noncytotoxic concentrations of the highly carcinogenic and mutagenic (-)-(1R,2S,3S,4R)-3,4 dihydroxy-1, 2-epoxy-1,2,3,4-tetrahydrobenzo[c]phenanthrene [(-)-B[c]PhDE; fjord region diol epoxide] or its biologically less active (+)-(1S,2R,3R,4S) enantiomer [(+)-B[c]PhDE]. The benzylic 4-hydroxyl group and the epoxide oxygen are trans in both enantiomers. Independent 8-azaguanine-resistant clones were isolated. The coding region of the hypoxanthine (guanine) phosphoribosyltransferase gene was amplified by reverse transcription-PCR and sequenced. For (-)-B[c]PhDE, mutation frequencies were 10- or 356-fold above background for the low (0.01-0.1 microM; 97% cell survival) or high (1.0-1.25 microM; 26% cell survival) doses, respectively. For the high dose group, 20 of 64 base substitutions occurred at GC base pairs (31%) and 44 at AT base pairs (69%). For the low-dose group, 6 of 55 base substitutions were at GC base pairs (11%), and 49 were at AT base pairs (89%). For the less active (+)-B[c]PhDE, mutation frequencies were 17- or 372 fold above background for the low (0.12-0.5 microM; 95% cell survival) or high (2.0-3.0 microM; 31% cell survival) doses, respectively. In contrast to the results with the (-)-B[c]PhDE, both the high- and the low-dose groups for (+) B[c]PhDE gave a 50:50 distribution of base substitution at GC versus AT base pairs. Our data indicate that: (a) transversions were the predominant base substitutions observed for both the (+)- and (-)-enantiomers of B[c]PhDE; (b) (-) B[c]PhDE showed high selectivity for causing AT --> TA transversions, whereas considerably less selectivity was observed for (+)-B[c]PhDE; (c) (-)-B[c]PhDE had a different hot spot profile for base substitutions than did (+)-B[c]PhDE, but some common hot spots were observed for both compounds; and (d) decreasing the dose of (-)-B[c]PhDE increased the proportion of mutations at AT base pairs and decreased those at GC base pairs, but this was not observed for (+)-B[c]PhDE. PMID- 8706011 TI - Phase II enzyme expression in rat liver in response to the antiestrogen tamoxifen. AB - The genotoxicity and carcinogenicity of tamoxifen have been attributed to metabolic activation of tamoxifen to an electrophile. Phase II enzymes are known to be involved in the metabolism of the drug and possibly in the formation or elimination of the active metabolite. To determine the effects of tamoxifen on phase II enzyme expression, the drug was administered to F344 rats, and hepatic glutathione S-transferase (GST), UDP-glucuronosyltransferase (UGT), and sulfotransferase (ST) expression was evaluated. Some of the tamoxifen-induced effects, including dramatic suppression of selected GST enzymes and activity, were observed at a dose in rats that is directly equivalent, on a mg/kg b.w. basis, to the doses used for breast cancer treatment. Most of the observed responses are not consistent with the previously described phenobarbital-like properties of tamoxifen and could be the result of the partial agonist activity of tamoxifen at the estrogen receptor. Northern blot analysis was performed with isozyme-specific oligonucleotide probes for rat GST, ST, and UGT. In addition, GST subunit protein levels were assayed by high-performance liquid chromatography. In females, tamoxifen treatment resulted in a 60% suppression of GST Ya1 mRNA and protein levels and a 40% suppression of GST Ya2 levels. In males, tamoxifen treatment suppressed GST Ya1 expression approximately 60%, and GST Ya2 expression was suppressed at low doses but induced above control at high doses. Male GST Yc1 was induced approximately 80% over control. The expression of all other major forms of rat hepatic GST subunit protein, including GST Yb1, Yb2, Yb3, Yp, and Yl, was unaffected by tamoxifen treatment. GST conjugation activity toward delta 5-androstene-3,17-dione, a GST Ya1- and Ya2-specific substrate, was suppressed approximately 40% in both sexes, consistent with our protein and mRNA data. Total GST activity, as measured by the rate of chlorodinitrobenzene conjugation, was not changed. Tamoxifen also produced a dose-dependent increase in UGT2B1 mRNA, a phenobarbital-inducible enzyme; mRNA levels reached 210 and 420% of control in females and males, respectively. In addition, mRNA levels for ST2A2, a female-specific ST gene, were suppressed 50% in females and induced 120% over control in males. mRNA expression for all other forms of rat liver UGT and ST isozymes that were tested was not significantly affected by tamoxifen treatment. Overall, these results demonstrate that tamoxifen has significant effects on hepatic phase II enzyme expression that may have implications for the carcinogenicity and/or therapeutic activity of the drug. PMID- 8706012 TI - A new process of cancer prevention mediated through inhibition of tumor necrosis factor alpha expression. AB - Mechanisms of cancer prevention were studied using structurally different cancer preventive agents, sarcophytol A, canventol, (-)-epigallo-catechin gallate, and tamoxifen, based on our evidence that tumor necrosis factor alpha (TNF-alpha) acts as an endogenous tumor promoter relevant to human carcinogenesis. Pretreatment with the four preventive agents commonly inhibited TNF-alpha mRNA expression and TNF-alpha release in BALB/3T3 cells induced by a tumor promoter, okadaic acid, whereas the expression of early response genes (c-jun, junB, c-fos, and fosB) was enhanced. These results strongly suggest that inhibition of TNF alpha mRNA expression and its release is a new process of cancer prevention. PMID- 8706013 TI - Two dominant host resistance genes to pre-B lymphoma in wild-derived inbred mouse strain MSM/Ms. AB - To explore possible host genes suppressing spontaneous B-lymphomagenesis in the mouse, expression of ecotropic murine leukemia virus (E-MuLV) and lymphoma development were observed in crosses between the pre-B lymphoma-prone SL/Kh and low-lymphoma strains of mice. E-MuLV expression was intensely inhibited in F1 hybrids with the strains either with the Fv-1b allele (BALB/C, C57BL/10, and A/J) or with the Fv-1nr allele (NZB). In these F1 mice, no lymphoma developed by 18 months of age. On the other hand, F1 hybrids with the strains with the Fv-1n allele [C3H/He, CBA/N, SJL, DBA/2, and MSM/Ms (hereafter referred to as MSM)], high or intermediate levels of E-MuLV expression were observed. Lymphoma incidence in these F1 hybrids, however, was low. This observation suggests the presence of non-Fv-1 dominant resistance genes in these strains. In an attempt to characterize such host genes, we analyzed crosses between SL/Kh mice and a wild mouse-derived inbred strain, MSM/Ms. The latter was susceptible to N-tropic virus expression, but (SL/Kh x MSM)F1 hybrids, did not develop and lymphomas. Of 60 SL/Kh x (SL/Kh x MSM)F1 hybrids, 14 B-lineage lymphomas, including 13 pre-B and 1 follicular center cell lymphoma, developed by 18 months of age. This was compatible with the hypothesis of two independently segregating dominant genes of MSM suppressing lymphomagenesis. By scanning all chromosomes for linkage of lymphoma susceptibility with polymorphic microsatellite loci, one significant linkage disequilibrium was found in the proximal segment of chromosome 17, containing D17MIT44 (map position 15.0) to D17MIT150 (position 33.3), and another linkage disequilibrium, in the midproximal segment of chromosome 18, containing D18MIT90 (map position 28.0) and D18MIT140 (37.0). All 13 pre-B lymphoma-bearing backcross mice were homozygous for SL/Kh-derived alleles at these loci. We named the gene on chromosome 17 Msmr1 (for MSM resistance 1) and that on chromosome 18 Msmr 2 (for MSM resistance 2). PMID- 8706014 TI - Increased resistance to N,N',N"-triethylenethiophosphoramide (thiotepa) in cells expressing the Escherichia coli formamidopyrimidine-DNA glycosylase. AB - Thiotepa (N,N',N'-triethylenethiophosphoramide) is an alkylating agent used in cancer chemotherapy. A reaction pathway by which thiotepa alkylates purified DNA involves hydrolysis to aziridine, which forms N7-aminoethyl guanine and aminoethyl adenine. These lesions are repaired in Escherichia coli by the formamidopyrimidine-DNA glycosylase (Fpg) protein. To determine whether such lesions are formed by thiotepa in mammalian cells, we have overexpressed the E. coli Fpg protein in Chinese hamster ovary cells. The transfected cells were more resistant to the lethal and mutagenic effects of thiotepa than the parental cells. The number of replication-blocking lesions formed by thiotepa, measured by quantitative PCR analysis, was lower in the transfected cells. These results show that expression of the Fpg protein increases the cell resistance to thiotepa and suggest that this compound produces ring-opened guanines, which are involved in its cytotoxic action. PMID- 8706015 TI - Prognostic significance of germ line polymorphisms of the CYP1A1 and glutathione S-transferase genes in patients with non-small cell lung cancer. AB - CYP1A1 is responsible for the metabolic activation of benzo(a)pyrene in cigarette smoke, and high susceptibility to smoking-related lung cancer has been associated with the MspI polymorphism of the CYP1A1 gene. Individuals with a susceptible CYP1A1 genotype have been found to be at remarkably high risk when the genotype is combined with a deficient Mu-class glutathione S-transferase (GSTM1) genotype. In this study, we investigated the relationship between germ line polymorphisms of these genes and clinical characteristics or survival rates in 232 patients with non-small cell lung cancer (NSCLC). Statistical analysis revealed a significant association (P < 0.05) of the MspI polymorphism of the CYP1A1 gene with histological type, performance status (general conditions of patients), and the extent of the primary tumor (T-factor). On the other hand, the GSTM1 polymorphism was significantly associated with performance status, the extent of regional lymph node metastasis (N-factor), and the extent of distant metastasis (M-factor). NSCLC patients with at least one susceptible allele of the MspI polymorphism of the CYP1A1 gene [heterozygous genotype B or a rare homozygous genotype C; n = 131; median survival time (MST) = 24.2 months] were associated with a shortened survival compared with those with nonsusceptible homozygous alleles (genotype A; n = 101; MST = 65.2 months; P = 0.005 by log-rank test). Smokers with susceptible genotypes (n = 104; MST = 18.2 months) were markedly associated with a shortened survival compared with those with genotype A (n = 76; MST = 69.2 months; P = 0.024); such an association was not found among nonsmokers by genotypes. Genotype-dependent survival was also observed in patients at an advanced stage of disease (P = 0.010), but not in those at an early stage of disease (P = 0.382). Patients with the susceptible CYP1A1 genotype had remarkably shortened survivals when the genotype was combined with a deficient genotype GSTM1(-) (P = 0.017; degree of freedom = 3). Multivariate analysis by the Cox proportional hazards model also revealed that the CYP1A1 polymorphism was an independent prognostic factor in patients at a nonresectable advanced stage of NSCLC (P = 0.005; hazard ratio = 1.98; 95% confidence interval, 1.24-3.17). PMID- 8706016 TI - Augmentation of antitumor immunity with bacterial superantigen, staphylococcal enterotoxin B-bound tumor cells. AB - We have examined the effectiveness of a bacterial superantigen, staphylococcal enterotoxin B (SEB)-coupled tumor cells, to induce antitumor activity. SEB was chemically conjugated to tumor cells using a heterobifunctional cross-linking agent acting through NH2 and SH groups. V beta 8+ T cells were activated and increased in number after the culture with SEB-bound Meth A cells. The cultured T cells exhibited an antitumor activity in the Winn assay. Interleukin 2 (IL-2) receptor alpha + (IL-2R+) V beta 8+ T cells but not IL-2R+ V beta 6+ T cells increased in number in mice injected with SEB-bound Meth A cells. However, the percentages of V beta 8+ and V beta 6+ T cells did not change by this immunization. The antitumor effector cells were V beta 7- 8- CD4+ T cells. In vivo immunization with SEB-bound cells induced a strong antitumor activity, i.e., tumor-free mice/total mice = 14 of 15 (93%) for Meth A and 7 of 15 (47%) for hepatoma MH134. The induced antitumor activity was both dose dependent and tumor specific. Treatment with SEB-bound cells prolonged the survival days of Meth A bearing mice by 62%. These results suggest that SEB-bound tumor cells may be a powerful method for induction of in vivo antitumor activity. PMID- 8706017 TI - Brain-derived neurotrophic factor protects neuroblastoma cells from vinblastine toxicity. AB - Brain-derived neurotrophic factor (BDNF) and its receptors are necessary for the survival and development of many neuronal cells. Because BDNF and TrkB are expressed in many poor-prognosis neuroblastoma (NB) tumors, we evaluated the role of BDNF in affecting sensitivity to chemotherapeutic agents. We investigated the effects of activation of the BDNF-TrkB signal transduction pathway in two NB cell lines, 15N and SY5Y. 15N cells lack the high-affinity receptor p145TrkB and express BDNF; 15N cells were used along with 15N-TrkB cells, a subline transfected with a TrkB expression vector. In cytotoxicity assays, 15N-TrkB cells were consistently 1.4-2 fold more resistant to vinblastine than 15N cells. Drug accumulation assays showed a 50% reduction in[3H]vinblastine accumulation in 15N TrkB cells compared with control 15N cells. Addition of 30 ng/ml BDNF resulted in a reduction to 46% of control in 15N cells and a reduction to 28% of control in 15N-TrkB cells. SY5Y cells were chosen as a second model because they lack both endogenous BDNF and TrkB expression. p145TrkB expression is induced by 1 nM retinoic acid. Vinblastine accumulation was not significantly affected by 1 nM retinoic acid in SY5Y cells. Addition of 30 ng/ml BDNF decreased [3H]vinblastine accumulation to 58% of control in SY5Y cells and decreased [3H]vinblastine accumulation to 62% of control in TrkB-expressing SY5Y cells. Although an increase in BDNF expression in seen in multidrug-resistant sublines of SY5Y and BE(2)-C NB cells, the protective effect of BDNF in vinblastine toxicity may be unrelated to mdr-1, because the activity of other agents transported by P glycoprotein was not affected. There was no increase in mdr-1 expression in 1 nM RA SY5Y cells and 15N-TrkB cells, as assessed by Northern blot analysis. In addition to the effects of BDNF on vinblastine cytotoxicity and accumulation, there was an inhibition in the ability of vinblastine to depolymerize tubulin in BDNF-treated cells. Thus, BDNF and TrkB may partially rescue NB cells from vinblastine toxicity and thereby may contribute to a more chemoresistant phenotype. PMID- 8706018 TI - Doxorubicin encapsulated in sterically stabilized liposomes is superior to free drug or drug-containing conventional liposomes at suppressing growth and metastases of human lung tumor xenografts. AB - Liposomes containing polyethylene glycol-derivatized phospholipids are able to evade the reticuloendothelial system and thereby remain in circulation for prolonged periods. We report here that doxorubicin encapsulated in these sterically stabilized liposomes (S-DOX) suppresses the growth of established human lung tumor xenografts in severe combined immunodeficient (SCID) mice and inhibits the spontaneous metastases of these tumors. The enhanced therapeutic efficacy of S-DOX compared to free doxorubicin was demonstrated in two independent human/mouse models. In the first model, S-DOX inhibited the growth of a human non-small cell lung tumor xenograft established orthotopically in the lungs of SCID mice. Treatment of these mice with S-DOX, but not with free drug, suppressed the growth of the tumor in the lung, prevented metastasis from the lung, and enhanced survival percentage. In another model, the human lung tumor is engrafted into gonadal fat pad of SCID mice. Human tumor xenografts grow floridly in this site of engraftment, and the tumor spreads from this primary site into the peritoneal cavity and subsequently reaches the liver and lung. In this model, free drug suppressed the growth of the primary tumor but had no effect upon the subsequent spread of the tumor into the peritoneal cavity, liver, and lung. In contrast, treatment of the tumor-bearing mice with S-DOX (but not with doxorubicin in conventional liposomes) suppressed the tumor spread to the peritoneal cavity, completely arrested metastasis to the liver and lung, and suppressed the growth of the primary tumor xenograft. This report provides the first evidence that antitumor drugs delivered by sterically stabilized liposomes can arrest the metastasis of human tumor xenografts. PMID- 8706019 TI - Up-regulation of vascular endothelial growth factor production by iron chelators. AB - Agents that modulate cellular iron availability have been studied for their antitumor activity. Based on encouraging in vitro studies, the iron chelator deferoxamine (DFO) has been used in clinical studies to treat cancer patients. The observation that DFO induced macular edema in several cancer patients led to the present investigation of vascular endothelial growth factor (VEGF) as a possible mediator of the encountered side effects. Both normal and malignant cell lines were incubated with DFO and a variety of other iron chelators. DFO, at concentrations achievable in humans, induced a 3-5-fold increase in VEGF mRNA expression in all cell lines studied. This increased VEGF mRNA expression was dose and time dependent. A panel of structurally different iron chelators induced an even more potent increase in VEGF mRNA expression. The DFO-induced increase in VEGF mRNA expression translated into 6- and 4-fold increases in VEGF protein secretion in conditioned media of retinal pigment epithelial and C6 glioblastoma cells, respectively. These findings suggest that VEGF may act as a mediator of the side effects induced by iron chelation therapy. In addition, because VEGF is an important regulator of angiogenesis, iron chelators should be given with caution to cancer patients. PMID- 8706020 TI - Involvement of beta-glucuronidase in intestinal microflora in the intestinal toxicity of the antitumor camptothecin derivative irinotecan hydrochloride (CPT 11) in rats. AB - Irinotecan hydrochloride (CPT-11), an antitumor camptothecin derivative, causes severe forms of diarrhea clinically. We characterized CPT-11-induced diarrhea histologically and enzymologically and assessed the relationships between intestinal toxicity and the activity of the enzymes that play a key role in the major metabolic pathway of CPT-11 in rats. CPT-11 (60 mg/kg i.v. for 4 days) induced intestinal toxicity characterized by severe chronic diarrhea, loss of body weight, and anorexia. Histological damage was most severe in the cecum. The segmental difference in the degree of the damage showed good correlation with the beta-glucuronidase activity in the contents of the lumen in each case, but not with the intestinal tissue carboxylesterase activity, which converts CPT-11 to its active form (7-ethyl-10-hydroxycamptothecin). Inhibition of the beta glucuronidase activity in the intestinal microflora by antibiotics (1 mg penicillin and 2 mg streptomycin per ml of drinking water) markedly ameliorated the diarrhea and reduced cecal damage. Analysis of CPT-11 and its metabolites in the feces indicated that antibiotics completely inhibited the deconjugation of the glucuronic conjugate of 7-ethyl-10-hydroxycamptothecin by beta-glucuronidase. It is suggested that CPT-11-induced diarrhea would be attributable to the damage to the cecum, and that the inhibition of the beta-glucuronidase activity in the intestinal microflora is a major protective effect of antibiotics. PMID- 8706021 TI - Combination suicide and cytokine gene therapy for hepatic metastases of colon carcinoma: sustained antitumor immunity prolongs animal survival. AB - The effectiveness of combination therapy using a suicide gene and cytokine genes for the treatment of metastatic colon carcinoma in the mouse liver was investigated. Pre-established hepatic tumors treated with a recombinant adenoviral vector containing the herpes simplex virus thymidine kinase gene(tk) exhibited substantial regression, although all treated animals suffered from subsequent relapses. Although cotreatment with a mouse interleukin 2 (mIL-2) containing adenoviral vector induced an effective antitumor immune response, the immunity waned with time, and the treated animals eventually succumbed to hepatic tumor relapse or distant metastases. In this study, mouse granulocyte macrophage colony-stimulating factor (mGM-CSF) gene was tested for its ability to further enhance and prolong the antitumoral cellular immunity. A fraction of the animals treated with tk + mIL-2 + mGM-CSF developed long-term antitumor immunity and survived for more than 4 months without recurrence. This long-term antitumor immunity could be enhanced further by subsequent "vaccination" with mIL-2 expressing parental tumor cells. The results indicate that local expression of GM CSF in the hepatic tumors and prolonged mIL-2 expression are necessary to generate persistent antitumor immunity that is essential for the prevention of tumor recurrence and long-term animal survival. PMID- 8706022 TI - Human dendritic cells genetically engineered to express high levels of the human epithelial tumor antigen mucin (MUC-1). AB - We have achieved stable high-level expression of a human tumor antigen, epithelial cell mucin (MUC-1), on human dendritic cells (DCs) by retroviral transduction of CD34+ progenitor cells and their subsequent cytokine-induced differentiation into DCs. The process of retroviral transduction did not alter the growth or differentiation of DCs from CD34+ progenitor cells. Immunofluorescence and electron microscopy studies revealed that the expression of mucin was limited to the body of the DCs and was excluded from the cytoplasmic veils of the DCs. Furthermore, the expression of mucin on DCs was similar, if not identical, to the nonpolarized expression of mucin found on carcinoma cells. In functional studies, the MUC-1(+)-transduced DCs were potent stimulators of allogeneic CD4+ T cells and, in fact, were superior to MUC-1- DCs. Thus, MUC-1+ DCs are expected to be a valuable tool in the immunotherapeutic treatment of patients with tumors that express MUC-1. PMID- 8706023 TI - Physiologically based kinetic model of effector cell biodistribution in mammals: implications for adoptive immunotherapy. AB - The goal of the present investigation was to develop a physiologically based kinetic model to describe the biodistribution of immunologically active effector cells in normal and neoplastic tissues of mammals based on the current understanding of lymphocyte trafficking pathways and signals. The model was used to extrapolate biodistribution among different animal species and to identify differences among different effector populations and between intra-arterial and systemic injections. Most importantly, the model was used to discern critical parameters for improving the delivery of effector cells. In the model, the mammalian body was divided into 12 organ compartments, interconnected in anatomic fashion. Each compartment was characterized by blood flow rate, organ volume and lymphatic flow rate, and other physiological and immunological parameters. The resulting set of 45 differential equations was solved numerically. The model was used to simulate the following biodistribution data: (a) nonactivated T lymphocytes in rats; (b) interleukin 2-activated tumor-infiltrating lymphocytes in humans; (c) nonactivated natural killer (NK) cells in rats; and (d) interleukin 2-activated adherent NK cells in mice. Comparisons between simulations and data demonstrated the feasibility of the model and the scaling scheme. The similarities as well as differences in biodistribution of different lymphocyte populations were revealed as results of their trafficking properties. The importance of lymphocyte infiltration from surrounding normal tissues into tumor tissue was found to depend on lymphocyte migration rate, tumor size, and host organ. The study confirmed that treatment with effector cells has not been as impressive as originally promised, due, in part, to the biodistribution problems. The model simulations demonstrated that low effector concentrations in the systemic circulation greatly limited their delivery to tumor. This was due to high retention in normal tissues, especially in the lung. Reducing normal tissue retention through decreasing attachment rate or adhesion site density in the lung by 50% could increase the tumor uptake by approximately 40% for tumor infiltrating lymphocytes and by approximately 60% for adherent NK cells. Our analysis suggested the following strategies to improve effector cell delivery to tumor: (a) bypassing the initial lung entrapment with administration to the arterial supply of tumor; (b) reducing normal tissue retention using effector cells with high deformability or blocking lymphocyte adhesion to normal vessels; and (c) enhancing tumor-specific capture and arrest by modifying the tumor microenvironment. PMID- 8706024 TI - Protective antitumor immunity induced by immunization with completely allogeneic tumor cells. AB - We have shown previously that immunization of B6 mice (H-2b) with tumor cells of B6 origin transformed by the human adenovirus type 5 early region 1 (Ad5E1) induces an H-2Db-restricted CTL response against an E1B-encoded CTL epitope. We now report that immunization of B6 mice with Ad5E1-transformed tumor cells of BALB/c origin (H-2d), apart from inducing a B6 anti-BALB/c allo-response, also induces a strong CTL response against the E1B-encoded H-2Db-presented CTL epitope. BALB/c Ad5E1-transformed tumor cells are not recognized by E1B-specific CTLs, indicating that nontumor cells have processed the E1B-encoded CTL antigen and have presented the E1B peptide to E1B-specific CTLs. These data also show that the B6 anti-BALB/c allo-response does not overwhelm the anti-E1B response induced by the allogeneic tumor cell vaccination. Moreover, B6 mice immunized with allogeneic BALB/c Ad5E1 cells are, in contrast to mice vaccinated with untransformed BALB/c cells, protected against a subsequent challenge with B6 Ad5E1-expressing tumor cells. These data show that immunization with completely allogeneic tumor cells can lead to protective syngeneic antitumor immunity, indicating that completely allogeneic tumor cell vaccines can be used for the induction of antitumor immunity. PMID- 8706025 TI - L-lysine effectively blocks renal uptake of 125I- or 99mTc-labeled anti-Tac disulfide-stabilized Fv fragment. AB - In this study, we investigated the ability of L-lysine to block renal uptake of 125I- or 99mTc- labeled Fv fragments. Anti-Tac disulfide-stabilized Fv fragment (dsFv) was derived from a murine monoclonal antibody that recognizes the alpha subunit of the interleukin-2 receptor (IL-2R alpha). The 125I- or 99mTc-labeled dsFv was injected i.v. into non-tumor-bearing nude mice or into nude mice bearing SP2/Tac (IL-2R alpha positive) and SP2/0 (IL-2R alpha negative) tumor. We then evaluated the pharmacokinetics of L-[3H]lysine and the effect of L-lysine dose, timing of administration, and route of delivery on catabolism and biodistribution of i.v. dsFv. Peak renal uptake of i.v. or i.p. injected L-[3H]lysine occurred within 5 and 15 min, respectively. The kidney uptake of L-lysine exhibited a dose response effect. When L-lysine was coinfused or injected shortly before dsFv, renal uptake of dsFv was blocked to < 5% of the control, but longer intervals were less effective. Aminosyn II and Travasol 10% (parenteral amino acid solutions) also blocked renal uptake of radiolabeled dsFv. Administration of L lysine did not alter the blood kinetics and slightly increased the tumor uptake of dsFv, but it did prevent catabolism in the kidney and resulted in lower amounts of catabolites in the serum and urine. In conclusion, we have shown that a blocking dose of lysine, injected with or immediately before the injection of radiolabeled dsFv, is most effective in blocking the renal uptake of dsFv. This is consistent with the rapid uptake of L-[3H]lysine by the kidney and is further substantiated by the relative ineffectiveness of lysine injected immediately after the radiolabeled dsFv injection. PMID- 8706026 TI - Selective inhibition of telomerase activity during terminal differentiation of immortal cell lines. AB - Telomerase, the enzyme that maintains the ends of linear eukaryotic chromosomes, is active in human germ cells and in a majority of tumor tissues and immortalized cell lines. In contrast, most mature somatic cells and tissues contain low or undetectable telomerase activity, implying a stringent negative regulatory control mechanism. We report here that telomerase activity is dramatically inhibited during the terminal differentiation of HL-60 human promyelocytic leukemia cells to monocytic and granulocytic lineages. A loss of telomerase activity was seen in response to three different inducers of differentiation, was independent of differentiation-induced apoptosis, and occurred in the presence of unaltered expression of the RNA component of telomerase. Reduction in telomerase activity was also observed during the differentiation of murine F9 teratocarcinoma and C2C12 myoblast cells. In contrast, induced differentiation of murine p19 embryonal carcinoma and Neuro 2a neuroblastoma cells did not result in a loss of telomerase activity. These results are therefore consistent with the absence of telomerase activity in human somatic cells and the presence of telomerase activity in many somatic murine cells and tissues. PMID- 8706027 TI - Mapping of chromosomal imbalances in pancreatic carcinoma by comparative genomic hybridization. AB - To identify recurrent chromosomal imbalances in pancreatic adenocarcinoma, 27 tumors were analyzed by using comparative genomic hybridization. In 23 cases chromosomal imbalances were found. Gains of chromosomal material were much more frequent than losses. The most common overrepresentations were observed on chromosomes 16p (eight cases), 20q (seven cases), 22q (six cases), and 17q (five cases) and under-representations on a subregion of chromosome 9p (eight cases). Distinct high-level amplifications were found on 1p32-p34, 6q24, 7q22, 12p13, and 22q. These data provide evidence for a number of new cytogenetically defined recurrent aberrations which are characteristic of pancreatic carcinoma. The overrepresented or underrepresented chromosomal regions represent candidate regions for potential oncogenes and tumor suppressor genes, respectively, possibly involved in pancreatic tumorigenesis. PMID- 8706028 TI - Chromosome 7q allelic losses in pancreatic carcinoma. AB - During our DNA fingerprinting studies of paired normal and pancreatic cancer tissues using arbitrarily primed PCR, we noticed a band showing an apparent homozygous deletion in a pancreatic cancer cell line and a decreased intensity in a number of primary cancers. That band was assigned to chromosome 7. Such information led us to analyze chromosome 7 loss of heterozygosity (LOH) in a panel of 12 cryostat-enriched primary pancreatic cancers and 2 pancreatic cancer cell lines, despite the reportedly low frequency of chromosome 7 LOH in xenograft enriched pancreatic cancers. Seventeen PCR-amplified CA-microsatellite polymorphic sites were analyzed. One of the two cell lines and eight common-type cancers (including all five poorly differentiated and three of five moderately differentiated cancers) showed chromosome 7q LOH, whereas the two uncommon types of ductal cancer (one adenosquamous and one mucinous noncystic) scored negative. Our data suggest that chromosome 7q LOH is a frequent event (80%) in cryostat enrichable common pancreatic ductal carcinomas, that is, those primarily of high cellularity. The chromosome 7q smallest common deleted region described by our cases was between 7q31.1 and 7q32. PMID- 8706029 TI - An uncertain role for p53 gene alterations in human prostate cancers. AB - Inactivation of the p53 gene has been implicated in prostate cancer progression. To determine the role of p53 inactivation in the progression of clinical prostatic carcinomas, we assessed 67 tumors derived from patients with clinically localized disease for chromosome 17p and p53 gene allelic loss, p53 gene mutations using single-strand conformational polymorphism and direct sequencing, and p53 protein expression using immunohistochemical staining. Of 55 informative tumors, 10 demonstrated loss of 17p or the p53 gene; however, only a single tumor had a mutation in its remaining p53 allele. Significant p53 overexpression was observed in 2 of 38 tumors, and 9 others had faint staining of a few nuclei ( < 1%). p53 overexpression occurred in no informative tumor with allelic loss or mutation. In a 1-7-year follow-up, positive immunohistochemical staining did not confer an increased risk of recurrence (risk of recurrence, 0.86, P = 0.78), whereas allelic loss of chromosome 17p appeared to be highly correlated with recurrence (risk of recurrence, 3.7, P = 0.003). In an unrelated group of 42 patients with metastatic prostate cancer, p53 overexpression was found in 26 tumors (62%), and 15(36%) had high grade staining. Neither the presence nor the degree of expression correlated with time to progression or time to death. This series suggests that p53 gene inactivation is rare in primary prostatic tumors, not essential to the development of prostate cancer metastases, and of limited use as a prognostic marker in patients with primary or metastatic disease. Another gene or genes on chromosome 17p may be involved in prostate cancer progression. PMID- 8706030 TI - Quantitative estimation of epidermal growth factor receptor and c-erbB-2 in human breast cancer. AB - Epidermal growth factor receptor (EGFR) expression by human breast cancer has been shown to predict poor patient outcome, as has amplification of the c-erbB-2 proto-oncogene. We have developed a quantitative immunohistochemical method for measuring protein levels of both receptors and have applied this to a series of 123 breast primaries. We find EGFR expression is substantially lower than normal in nearly all breast cancers (97%). Quantification of p185erbB-2 indicates overexpression in 91% of the tumors. Two separate tumor populations are apparent with levels of c-erbB-2 expression ranging from 0.33 to 19 and 45 to 480 times normal, respectively. Within the lower population, p185erbB-2 expression is inversely related to EGFR expression (rank correlation, P < 0.0005). Using fluorescent in situ hybridization we show that tumors in the latter population have c-erbB-2 amplification and that amplification is restricted to this group. Our findings indicate that significant overexpression of p185erbB-2 occurs in the absence of amplification; these lower levels of expression may have functional significance. Fifty-three patients underwent in vivo bromodeoxyuridine labeling, allowing flow cytometric analysis of tumor cell cycle kinetics. EGFR expression correlates directly to the labeling index (P = 0.011) and indirectly to potential doubling time (P = 0.010), but not to the duration of the S-phase (P = 0.502). Conversely, p185erbB-2 expression does not relate to indices of proliferation. Our results have important implications for the use of both receptor types as therapeutic targets. PMID- 8706031 TI - Increased expression of the insulin-like growth factor I (IGF-I) receptor gene in hepatocellular carcinoma cell lines: implications of IGF-I receptor gene activation by hepatitis B virus X gene product. AB - Hepatitis B virus infection is associated with acute and chronic liver disease and the development of hepatocellular carcinoma (hcc). Several lines of evidence have suggested that hepatitis B virus X protein (HBx), which is a transcriptional trans-activator, plays a role in the process of liver carcinogenesis. We have investigated the expression of insulin-like growth factor I (IGF-I) receptor in human hepatocellular carcinoma cell lines using SNU368 cells containing HBx and SNU387 cells, which lack HBx gene transcript (J-G. Park et al., Int. J. Cancer, 62: 276-282, 1995), in an attempt to understand its possible relationship to the HBx-induced hcc. The binding of 125I-labeled IGF-I to the SNU368 cells was 5-fold higher than that of SNU387 cells. The Scatchard analysis of the binding data revealed a single class binding site for IGF-I with Kd of 7.6 and 8.8 nM and maximum binding capacities of 169 and 33 fmol/10(5) cells, respectively. Therefore, the difference observed in 125I-labeled IGF-I binding between SNU368 and SNU387 cells was due to an increase in the number of IGF-I binding sites with no change in affinity for the IGF-I receptor. An enhanced level of IGF-I receptors in SNU368 cells was also observed by fluorescence-activated cell sorting analysis using a monoclonal antibody against human IGF-I receptor, alpha IR3. The level of IGF-I receptor RNA and the basal IGF-I receptor gene promoter activity in SNU368 cells were 5 and 10 times higher than those observed in SNU387 cells, respectively. To substantiate further that HBx could transactivate the expression of the endogenous IGF-I receptor gene, Hep G2 cells were transiently transfected with a HBx expression vector. The transfection of Hep G2 cells with an HBx expression vector resulted in increased levels of IGF-I receptor RNA, promoter activity, and 125I-labeled IGF-I binding by 2.6-, 2.8-, and 2-fold, respectively. As a result of higher levels of IGF-I receptor, the mitogenic effect of IGFs (IGF-I and IGF-II) on SNU368 cells was 6 times higher than that of SNU387 cells. These results suggest that HBx may play a role in the process of hcc by activating IGF-I receptor gene expression. PMID- 8706032 TI - Identification of novel regions of deletion in familial Wilms' tumor by comparative genomic hybridization. AB - Wilms' tumor, an embryonic renal neoplasm diagnosed primarily in young children, can occur in either a noninheritable (sporadic) or a familial form, with the latter presenting earlier and more often at bilateral sites. Although familial Wilms' tumor is thought to develop through inherited and acquired mutational inactivation of the two alleles of predisposing tumor suppressor genes, only a small percentage of cases can be accounted for by mutations affecting the WT1 gene or linkage to the Beckwith-Weidemann syndrome of the BWS region on the short arm of chromosome 11. To find chromosomal regions that might contain genes important in the development of this disease, we used comparative genomic hybridization to analyze tumor specimens from familial cases for chromosomal regions that were consistently lost. Although inherited lesions of tumor suppressors are most often inactivating point mutations, accompanying somatic lesions in the malignant clones are often chromosomal deletions; therefore, consensus regions of loss in familial tumors are likely to harbor genes linked to familial predisposition. There were extensive genomic aberrations among the eight familial cases studied, with an average of 6.5 changes/tumor (range, 0-22). The most consistent findings with likely biological relevance were deletions of chromosomes 4 (consensus, 4q21-qter), 9 (consensus, 9p21-pter), 20p, and 3 (consensus, 3q12-q21). These regions have not been previously implicated in Wilms' tumor and may harbor novel genes that could aid attempts to understand the familial predisposition as well as the development and progression of these tumors. PMID- 8706033 TI - Spontaneous intestinal carcinomas and skin neoplasms in Msh2-deficient mice. AB - Hereditary nonpolyposis colorectal cancer is associated with defects in DNA mismatch repair. Here, we characterize tumor susceptibility of the recently described Msh2-deficient mouse model. Within the first year of observation, all homozygous mice succumbed to disease, with lymphomas observed in at least 80% of the cases. The majority (70%) of animals 6 months or older developed intestinal neoplasms associated with APC inactivation. Microsatellite instability was more common in carcinomas than in adenomas, but uncommon in normal tissues. Some animals (7%) developed a variety of skin neoplasms analogous to the Muir-Torre syndrome. Msh2-/- mice implicate a direct role for mismatch repair in several neoplasms with striking phenotypic similarities to humans. PMID- 8706034 TI - Preliminary report: the short-term effects of direct p53 DNA injection in primary hepatocellular carcinomas. AB - A pilot study, to assess the therapeutic potential of percutaneous injection of wild-type p53 (wt-p53) in five patients with primary hepatocellular carcinoma is reported. Three of the five patients showed objective tumor response with reduction of the tumor volume on computed tomographic (CT) scan measurements as well as a significant fall of serum alphafetoprotein. Much further work will be needed to elucidate the mechanism of action. PMID- 8706035 TI - Use of gene marking in bone marrow transplantation. AB - We have used gene marking to investigate the mechanism of relapse and biology of reconstitution following bone marrow transplantation (BMT). The rationale for our initial protocols was to learn if residual malignant cells in autologous marrow contribute to subsequent relapse. Marked malignant cells were found at the time of relapse in 6/8 patients relapsing after autologous BMT for AML or neuroblastoma showing the infused marrow contributed to disease recurrence. Modifications of this marker approach with two distinguishable vectors are now being used to compare the efficacy of purging techniques. We were also able to evaluate gene transfer to normal progenitors and demonstrated that the marker gene was expressed for up to 36 months. Gene marking is also being used to trace the fate of EBV-specific CTLs that we are administering to recipients of allogeneic BMT and has provided evidence of persistence of adoptively transferred CTL for up to 10 months. PMID- 8706036 TI - Assessment of major carcinogenic tobacco-specific N-nitrosamines in Thai cigarettes. AB - This report presents new findings on the content of cancer-causing tobacco specific N-nitrosamines (TSNA) in mainstream smoke of nine brands of commercially produced Thai cigarettes, representing about 85% of market share in Thailand. Also tested were two major and popular brands of U.S. cigarettes sold in Thailand, representing about 10% of market share. The cigarettes included filter and nonfilter cigarettes with high, moderate, and low tar and nicotine yields. The observed range for N-nitrosonornicotine (NNN) was from 28 to 730 ng/cigarette and for 4-(methylnitrosamino)-1-(3-pyridyl-1-butanone (NNK) from 16 to 369 ng/cigarette. The relatively highest TSNA values were obtained in filter and nonfilter cigarettes with high tar (22.3-28.1 mg/cigarette) and high nicotine (1.78-2.42 mg/cigarette) deliveries. The results demonstrated that there is a correlation between TSNA and tar and nicotine deliveries in mainstream smoke. The TSNA deliveries, along with the tar and nicotine levels in mainstream smoke depended on the tobacco composition. According to these results, the tar levels alone, while significant, are not a sufficient measure for the biological activity and the carcinogenic potential of cigarettes in Thailand. Consumption of tobacco products nearly quadrupled over the last three decades (1966-1995) in Thailand. Lung cancer is the leading malignancy for men and a common malignancy for women in Thailand. This report provides information that may prove helpful in evaluating the TSNA-carcinogens burden on smokers. Our goal is to offer the scientific basis for voluntary and/or government-regulated reduction of the smoke yields of TSNA in tobacco products in Thailand and in other countries. PMID- 8706037 TI - Risk factors for breast cancer mortality among the National Breast Screening Study of Canada participants. AB - We studied the relationship between risk factor information and breast cancer mortality by means of a case control study, nested within the population of the National Breast Screening Study of Canada (NBSS). The characteristics of women aged 40-59 years, identified at the initial screen, who subsequently died of breast cancer up to 7 years from the initial screen, were compared with those of controls drawn from the same population. Among the factors evaluated in this study, number of live births and presence of symptoms in the breast revealed on direct questioning were found to be significantly related to breast cancer mortality. The results suggest a decrease in risk of dying of breast cancer associated with one or more live births (OR: 0.79, 95% CI: 0.68, 0.93), and an increase in risk of dying of breast cancer associated with symptoms in the breast revealed on direct questioning at the initial screen (OR: 2.01, 95% CI: 1.20, 3.37). PMID- 8706038 TI - Activity of the DNA repair protein O6-methylguanine-DNA methyltransferase in human tumor and corresponding normal tissue. AB - The DNA repair protein O6-methylguanine-DNA methyltransferase (MGMT) is a main determinant of resistance of cells towards the cytostatic effect of N nitrosoureas. Here we report the activity of MGMT in different types of human tumors. Colorectal tumors showed the highest mean of MGMT activity (481 +/- 258 fmol/mg protein) followed by ovarian tumors (437 +/- 291 fmol/mg), breast (306 +/ 204 fmol/mg), testicular (299 +/- 179 fmol/mg), and brain tumors (55 +/- 44 fmol/mg). Only in breast and brain tumors were MGMT-deficient samples found, with a frequency of 3 and 5%, respectively. No significant difference in mean MGMT activity was observed between breast tumors of grading I, II, and III. For brain tumors there was a tendency of MGMT to decline with increasing grading. Breast and colorectal carcinomas were compared with the corresponding normal tissue obtained from the same patient. The data showed for 11 out of 12 pairs of breast tissue and for 13 out of 15 pairs of colorectal tissue that MGMT activity in the tumor was equal to or, more frequently, higher than the activity found in the corresponding normal tissue. Thus, it appears that in these groups of tumors, increase of MGMT activity during tumor formation and progression is the rule rather than the exception. PMID- 8706039 TI - Inhibition of hepatocellular carcinoma development and erythrocyte polyamine levels in ODS rats fed on 3'-methyl-4-dimethylaminoazobenzene by hemicalcium ascorbate, 2-O-octadecylascorbic acid, and ascorbyl palmitate. AB - We examined the modifying effect of hemicalcium ascorbate (Ca-Asc), and its lipophilic derivatives, 2-O-octadecylascorbic acid (CV-3611) and ascorbyl palmitate (AscP), on hepatocarcinogenesis by 3'-methyl-4-dimethylaminoazobenzene (3'-Me-DAB) in ODS rats (a mutant unable to synthesize ascorbic acid). Male 14 week-old ODS rats were given a modified AIN-A diet or the diet containing 0.06% 3'-Me-DAB, and drinking water containing 0.1% ascorbic acid. Rats were divided into the following eight groups: Group 1, no treatment (basal diet alone); Group 2, Ca-Asc; Group 3, CV-3611; Group 4, AscP;Group 5, 3'-Me-DAB; Group 6, 3'-Me-DAB + Ca-Asc; Group 7, 3'-Me-DAB + CV-3611; and Group 8, 3'-Me-DAB + AscP. Ca-Asc (2 g/kg), CV-3611 (0.2 g/kg), and AscP (0.6 g/kg) was administered once every day by gavage. 3'-Me-DAB was given in the basal diet. After 17 weeks, animals were killed by exsanguination, and the liver was weighed and processed for histological examination. Treatment by CV-3611 exerted a marked inhibitory effect on the development of 3'-Me-DAB-induced hepatocellular carcinomas (HCC) as measured by multiplicity. Although less effective than CV-3611, Ca-Asc and AscP also showed inhibitory effect. We have also studied the correlation of erythrocyte (RBC) polyamine levels and HCC development. RBC polyamine levels were inhibited by Ca-Asc and its derivatives, indicating it may be a marker of hepatocarcinogenesis. PMID- 8706040 TI - Effect of gonadotropin-releasing hormone analogs and their conjugates on gonadotropin-releasing hormone receptor--positive human cancer cell lines. AB - The purpose of the present investigation was to develop new gonadotropin releasing hormone (GnRH) antagonists and to increase their stability and antitumor effect by conjugation with carrier macromolecules. Antitumor effect was evaluated using clonogenic assay, cell counting for antiproliferation, and sulforhodamine B method. The presence of GnRH-binding sites in human cancer cell lines (MCF-7, MDA-MB-231, Ishikawa, LNCaP) was proved. The direct growth inhibition of tumor cell lines is achieved with relatively high analog concentrations (10(-10)- 10(-5) M). We have developed new GnRH analogs of human and chicken origin. MI-1544 (Ac-D-Trp1,3,D-Cpa2,D-Lys6,D-Ala10)GnRH and the chicken GnRH antagonist MI-1892 (Ac-D-Trp1,3, D-Cpa2, Lys5, [beta-Asp(DEA)]6, Gln8, D-Ala10)-GnRH have stronger direct antitumor properties than the agonists. The antagonists inhibited proliferation of GnRH receptor-positive human cancer cell lines by 28 to 38%. GnRH peptide analogs were coupled with macromolecules through biodegradable groups, to enhance their antitumor effects. The antagonists reduced survival of MCF-7 and MDA-MB-231 cells by 38 to 48% and 20 to 41%, respectively. They showed less activity against human endometrial and prostate cancer cells (10-20%). The copolymer (P) as polyanionic carrier molecule reached only 15 to 20% survival reduction in all cell lines. However, the copolymer GnRH antagonist conjugates P-X-1892 and P-X-1544 killed 95 to 98% of cells at doses corresponding to the GnRH analog concentration. These compounds having antitumor activity could be tried for the treatment of prostate, breast, and endometrium cancer. PMID- 8706041 TI - In vivo studies of the new gonadotropin-releasing hormone antagonist--copolymer conjugates having antitumor activity. AB - The aim of the study was to test in vivo the gonadotropin-releasing hormone (GnRH) antagonists and their conjugates showing antitumor activities in vitro. The in vivo experiments with the human GnRH antagonist MI-1544 (Ac-D-Trp1,3,D Cpa2,D-Lys6,D-Ala10)-GnRH, the chicken GnRH antagonist MI-1892 (Ac-D-Trp1,3,D Cpa2,Lys5,/beta-Asp(DEA)/6,Gln8,D-Al a10)-GnRH, and their copolymer conjugates were carried out on MCF-7 and MDA-MB-231 human breast tumors xenografted in immunosuppressed CBA/Ca HRIJ-T6 female mice and on MXT mouse mammary tumors in BDF1 mice. The P-X-1544 and P-X-1892 conjugates were prepared by coupling the GnRH antagonists to macromolecule copolymer through biodegradable spacers. MI 1544 and its conjugate had strong, whereas MI-1892 and its conjugate had slight, castration effect in rats. All of them showed selective antitumor activity. The conjugates, given daily, inhibited both types of xenografts by 42 to 49%. Their activity was stronger in MXT mammary tumor (72 to 61%). The in vivo effect of GnRH antagonists was largely increased by coupling them to nonbiodegradable macromolecule carriers of polyanionic character. P-X-1544 and P-X-1892 GnRH antagonist-macromolecule conjugates might become important therapeutic agents for the treatment of breast cancer. PMID- 8706042 TI - Effects of alprazolam on the development of Moloney sarcoma virus-induced tumors in stressed mice. AB - Experiments were conducted to evaluate the effects of alprazolam (1 mg/kg i.p.) on the development of autochthonous tumors induced by the Moloney sarcoma virus (MSV) in BALB/c female mice subjected to stress. Enhancement of MSV-induced tumor incidence and growth was observed, together with a delay in the usual prompt regression of the tumors, when programmed white sound anxiety-stress was administered immediately following MSV i.m. inoculation. However, a depressant effect on tumor size and incidence was observed when stress was administered before virus inoculation. Treatment with alprazolam was found to reverse partially the adverse effects of postinoculation stress, and also to inhibit the beneficial effects of the preinoculation administration of stress on tumor development. Pretreatment with Ro 15-1788 (10 mg/kg s.c.), a central benzodiazepine antagonist, resulted in a suppression on both effects of alprazolam in stressed mice. PMID- 8706043 TI - Retrospective epidemiologic analysis of patients diagnosed with intracranial meningioma from 1977 to 1990 at the Jackson Memorial Hospital, Sylvester Comprehensive Cancer Center: the Jackson Memorial Hospital Tumor Registry experience. AB - A descriptive analysis was performed of the Tumor Registry data for intracranial meningioma by the Jackson Memorial Hospital, University of Miami School of Medicine Tumor Registry. A total of 108 cases of intracranial meningioma was collected and reviewed. Overall survival for 2, 5, and 10 years was 82, 72, and 60%, respectively. There was no difference in survival for males and females at 5 years, nor any difference in survival for race or ethnicity. There was a trend for improved survival for the young age group (18-55 years). PMID- 8706044 TI - Genetic instability as a basis for transformation of human lymphocytes infected with human retrovirus. AB - T cell lines (Coculture-14, Coculture-5) derived from human T-cell leukemia virus type I (HTLV-I)-seronegative persons acquired interleukin-2 (IL-2)-dependent continuous growth capacity (immortalized) following in vitro HTLV-I infection. They showed structural abnormalities of chromosomes carrying proviral DNA as seen by in situ hybridization. Following ultraviolet (UV) irradiation, Coculture-5 cells achieved IL-2-independent autonomous growth (transformed) resulting in the establishment of UV-1 and UV-5 lines. They showed additional abnormalities of the same chromosomes. Cocultivation of Coculture-5 cells with IUdR-treated UV-1 cells also resulted in autonomous growth of Coculture-5 cells, giving rise to three cell lines. By ABC immunostaining with specific antibodies, expression of proteins coded for growth regulatory genes, including Ki-67, Topo II, Pol alpha, c-MYC, p53, Rb, bcl-3, bcl-2, and BM-1, was found to be variably altered in transformed cells compared with immortalized cells. These results demonstrated chromosomal instability, altered gene product expression of HTLV-I-infected human lymphocytes, and their susceptibility to transformation without exposure to an initiating carcinogen. PMID- 8706045 TI - The oncofetal protein p65: a new member of the steroid/thyroid receptor superfamily. AB - The 65-kDa oncofetal protein (p65), a potential tumor marker discovered and characterized in our laboratory, is highly conserved in different species. Its amino acid composition, peptide map, and N-terminal and internal peptide sequences are very similar if not identical in humans and rodents. We have now identified the p65 gene as a novel member of the superfamily of genes that encode nuclear receptors for various hydrophobic ligands such as steroids, vitamin D, retinoic acid, and thyroid hormones. these receptors are composed of several domains important in hormone binding, DNA binding, dimerization, and transcription activation. The human p65 cDNA was partially cloned, revealing at its C-terminal end regulatory elements typical of this superfamily of genes. The DNA-binding domain coincides with the cysteine-rich region encompassing the two conserved zinc fingers. In addition, the domain homologous to the receptor dimerization site was found close to the C-terminal end. The p65 protein is highly homologous to estrogen receptor in its DNA-binding domain but not in other regions of the sequence, indicating that p65 is a new receptor with an as yet unknown ligand. In addition, we have identified in the cloned p65 cDNA fragment sequences encoding two peptides, obtained by CNBr cleavage, whose amino acid sequences were previously established. PMID- 8706046 TI - Melanoma and vitiligo: immunology's Grecian urn. PMID- 8706047 TI - Protective effect of an acidic glycoprotein obtained from culture of Chlorella vulgaris against myelosuppression by 5-fluorouracil. AB - An acidic glycoprotein prepared from a culture of Chlorella vulgaris (CVS) was examined for its protective effect on 5-fluorouracil(5FU)-induced myelosuppression and indigenous infection in mice. Subcutaneous administration of CVS greatly reduced the mortality of non-tumor-bearing mice given a high dose of 5FU, and could increase the LD50 value of 5FU for these mice. After 5FU treatment, indigenous infection developed probably as a result of the impairment of the host defense system. CVS reduced the incidence of indigenous infections and this effect was attributable to the acceleration of recovery from 5FU-induced myelosuppression. Early recovery of hematopoietic stem cells, or cells responding to interleukin-3 or granulocyte/macrophage-colony-stimulating factor, was especially observed in the bone marrow of CVS-treated mice on days 4-9 after the injection of 5FU. When tumor-bearing mice were given CVS during treatment with 5FU, CVS prolonged the survival of mice without affecting the antitumor activity of 5FU. In addition, CVS was itself shown to exert an antitumor effect. These results suggested that CVS may be beneficial for the alleviation of side-effects in cancer chemotherapy without affecting the antitumor activity of the chemotherapeutic agent. PMID- 8706048 TI - Induction of gene expression for nitric oxide synthase by immunomodulating drugs in the RAW264.7 murine macrophage cell line. AB - We have elucidated the direct effects of PSK (a protein-bound polysaccharide) and OK-432 (a streptococcal preparation), both immunomodulating drugs, on the gene expression for an inducible nitric oxide synthase and on the production of nitric oxide (NO) in the RAW264.7 murine macrophage cell line. As determined by northern blot analysis, both immunomodulating drugs were potent inducers of gene expression for inducible NO synthase when cells were costimulated with interferon gamma (IFN gamma). Expression of mRNA for the enzyme occurred in a dose-dependent manner after 3 h, when 10-50 micrograms/ml PSK or 0.001-1 KE/ml OK-432 was used. Furthermore, NO was also produced in response to these drugs, as detected by the Griess reagent reaction. The enhancement of NO synthesis was thought to be mediated, in part, through tumor necrosis factor alpha (TNF alpha) induction by these agents, since a neutralizing antibody to TNF alpha significantly suppressed NO production in RAW264.7 cells stimulated with PSK or OK432 in combination with IFN gamma. We speculate that NO production may play a role in tumoricidal and microbicidal activities of PSK or OK-432 in vivo. PMID- 8706049 TI - Bacillus Calmette-Guerin plus interleukin-2 and/or granulocyte/macrophage-colony stimulating factor enhances immunocompetent cell production of interferon-gamma, which inhibits B16F10 melanoma cell growth in vitro. AB - Although immunotherapy with bacillus Calmette Guerin (BCG) is an established adjuvant treatment for malignant melanoma, the mechanism of its role in this process is unclear. To investigate the possible contribution of tumor-inhibitory cytokines induced by BCG, B16F10 melanoma cell growth in culture was assessed in response to purified cytokines and conditioned media of BCG-stimulated splenocytes. Interferon-gamma (IFN gamma) was the most potent single agent (IC50 approximately 50 pg/ml). Tumor necrosis factor alpha was substantially weaker (IC50 > 10 ng/ml) but provided synergy with IFN gamma. None of the other cytokines such as interleukin-2 (IL-2), IL-4, IL-6, IL-10, IL-12, or granulocyte/macrophage-colony-stimulating factor had direct antitumor activity against B16F10 melanoma cells. However, when IL-2 and/or GM-CSF were combined with BCG either by exogenous addition or through endogenous production by novel cytokine-secreting recombinant BCG (rBCG), a substantial increase in INF gamma production by splenocytes was observed. Antitumor activity of this conditioned medium directly correlated with IFN gamma concentration and was completely blocked by neutralizing antibody to IFN gamma. These results suggest that BCG may exert part of its antitumor action on melanoma through the induction of IFN gamma, which can be greatly enhanced through the concomitant addition of IL-2 and/or GM-CSF. Furthermore, by utilizing rBCG that secrete these cytokines, it may be possible to potentiate the antitumor effect of BCG directly at the site of BCG inoculation. PMID- 8706050 TI - Suppressive effects of Lactobacillus casei cells, a bacterial immunostimulant, on the incidence of spontaneous thymic lymphoma in AKR mice. AB - The mean survival age of female AKR/J mice was significantly prolonged, the enlargement of thymus was markedly suppressed, and the proliferation of ecotropic and recombinant murine leukemia viruses (MuLV) was markedly inhibited when 8-week old female AKR/J mice were injected intraperitoneally (i.p.) with heat-killed Lactobacillus casei cells twice weekly for 8 weeks. In contrast, such actions of heat-killed L. casei cells were not seen in 20-week-old female AKR/J mice. The leukemogenic activity of the cell-free extract of thymus from adult female AKR/J mice in newborn female AKR/J mice was drastically reduced by i.p. treatment with heat-killed L. casei cells. The difference in adjuvant effectiveness of heat killed L. casei cells on 8- and 20-week-old animals may be dependent on the difference in the enhancing activity of the cell-mediated immune systems between the groups induced by heat-killed L. casei cells, and, as a result, on the difference in the degree of proliferation of ecotropic and recombinant MuLV in thymus, which consequently causes thymic lymphoma. PMID- 8706051 TI - 99mTc-CD19 monoclonal antibody is not useful for imaging of B cell non-Hodgkin's lymphoma. AB - In this study we investigated the applicability of 99mTc-labeled CD19 monoclonal antibody (mAb) for tumor imaging in patients with B cell non-Hodgkin's lymphoma. A 1-mg sample of murine CD19 mAb was labeled with approximately 550 MBq [99mTc]pertechnetate. The labeled mAb was administered i.v. to seven patients, four without and three with pretreatment with 10 mg unlabeled CD19 mAb. The number of circulating B cells was decreased by 44 +/- 5% 1 h after injection of the radiolabeled mAb. Peripheral B cells were coated with CD19, resulting in partial modulation of CD19, most pronounced in the three pretreated patients. Whole-body images were obtained with a gamma camera and compared with results obtained by conventional imaging techniques. Initially, blood-pool activity dominated, whereas 24 h after injection the radioactivity was mainly located in the spleen, kidneys and liver. In two patients, a lesion in the spleen appeared as an unlabeled spot. In one patient, a lesion in the femur, which was detected by computed tomography (CT) and gallium-67 scans, was also seen on the CD19 scan from 1 h after administration of the radioimmunoconjugate onwards. Good imaging of bone marrow infiltration was observed in one of three patients. Lymph node involvement was not observed in any of the patients in whom affected lymph nodes were detected by CT or gallium-67 scan. In conclusion, in the present study radioimmunodetection with 99mTc-labeled CD19 mAb was found to be inferior to CT and gallium-67 scanning in the diagnosis of patients with B cell non-Hodgkin's lymphoma. PMID- 8706052 TI - Anti-tyrosinase antibodies in malignant melanoma. AB - Anti-tyrosinase antibodies were measured by enzyme-linked immunosorbent assay in sera of patients with malignant melanoma with either metastatic disease or no evidence of disease, in patients with melanoma and associated hypopigmentation (MAH), in patients with vitiligo and in healthy volunteers. The mean relative absorbance (Arel) was calculated by dividing the absorbance of each sample by the mean value for the control group. Using this method, the Arel of the control group was 1.000(SE 0.083). Arel of patients with metastatic disease (1.516; SE 0.225) was significantly higher (P = 0.03) than the value for the controls, but insignificantly higher than that for patients with no evidence of disease (1.216; SE 0.148). Patients with no evidence of disease, in whom the primary lesion originated in the lower limb, had a significantly higher (P = 0.01) Arel than the healthy volunteers. Patients with metastatic disease showed higher Arel if their primary lesions were confined to the area of the head and neck or to the lower limb. Patients with vitiligo had higher Arel values for their anti-tyrosinase antibody than any of the other groups. However, those with melanoma and MAH (vitiligo-like) had the same Arel of anti-tyrosinase antibodies as the controls or the patients with metastatic melanoma. This observation reflected the possible absorption of anti-tyrosinase antibodies to melanoma antigens, and pointed to the participation of anti-tyrosinase antibodies in the destruction of normal melanocytes in patients with melanoma, as part of the immune reaction towards this disease. PMID- 8706053 TI - Pre-immunotherapy serum CA27.29 (MUC-1) mucin level and CD69+ lymphocytes correlate with effects of Theratope sialyl-Tn-KLH cancer vaccine in active specific immunotherapy. AB - Patients with metastatic breast, colorectal or ovarian cancers received active specific immunotherapy (ASI) with Theratope sialyl-Tn-KLH (keyhole limpet hemocyanin) cancer vaccine emulsified in Detox adjuvant. The median log2 anti-STn IgG titer generated by ASI, estimated by enzyme-linked immunosorbent assay with solid-phase ovine submaxillary mucin, was 5.322 (range = 0 - 9.322). Following ASI, 51 patients who generated titers higher than the median value for anti-STn+ mucin IgG survived longer than 46 patients who generated lower titers below the median. 38 of the patients were phenotyped for CD69 prior to ASI. The patients with lower numbers of CD69+ peripheral blood lymphocytes prior to immunotherapy (pre-ASI) also had low serum CA27.29 cancer antigen (MUC-1) levels, and had longer times to disease progression and improved survival following ASI. Elevated pre-ASI serum CA27.29 tumor antigen levels were associated with higher numbers of CD69+ PBL, with decreased anti-STn antibody production and decreased survival following ASI. The data are compatible with the hypothesis that elevated serum MUC-1 mucin is specifically immunosuppressive. PMID- 8706055 TI - ["Significance" of coronary stenosis]. PMID- 8706054 TI - Effect of intraperitoneal administration of granulocyte/macrophage-colony stimulating factor in rats on omental milky-spot composition and tumoricidal activity in vivo and in vitro. AB - Milky spots in the greater omentum are small accumulations of leucocytes that consist mainly of macrophages and have recently shown to be a selective dissemination site of intraperitoneal (i.p.) inoculated tumour cells. However, milky-spot macrophages show tumoricidal activity and may, therefore, be an excellent source of effector cells suited for local immunotherapy. In the present study we first examined whether granulocyte/macrophage-colony-stimulating factor (GM-CSF) treatment of isolated milky-spot macrophages affects the cytotoxicity against syngeneic colon carcinoma cells (CC531) in vitro. Secondly, we studied the influence of intraperitoneal GM-CSF administration on the number and antitumour activity of milky-spot and peritoneal macrophages. All studies were performed in Wag/Rij rats in which a syngeneic colon carcinoma cell line (CC531) is available. The results of the in vitro study showed that GM-CSF treatment of the omental macrophages led to an increased cytotoxicity against the tumour cell line. Intraperitoneal administration of 1000 U GM-CSF daily for 7 consecutive days demonstrated both an enhanced antitumour activity of the milky-spot macrophages and an increase in the milky-spot macrophage population. An increase in the proliferative capacity, according to bromodeoxyuridine incorporation, was shown in the milky-spot macrophages. Taking into account both the enhanced macrophage number and their enhanced activity upon i.p. GM-CSF treatment, the milky-spot macrophages may provide a rationale for local intraperitoneal immunotherapy in the prevention of intra-abdominal tumour growth. PMID- 8706056 TI - [Left ventricular remodeling after myocardial infarction]. PMID- 8706057 TI - Right sided pulsus alternans: a neglected phenomenon. PMID- 8706058 TI - [The utility of electrocardiography in the age of exercise echocardiography]. PMID- 8706059 TI - Effects of lung water on respiratory mechanics. PMID- 8706060 TI - The place of beta-blockers in cardiovascular medicine. PMID- 8706061 TI - [Thrombophilic state inpatients suffering from myocardial infarction with or without carotid atherosclerotic lesions]. AB - The aim of the present study was to evaluate some metabolic, coagulation and fibrinolytic parameters in 35 patients (24 males and 11 females, mean age 57 +/- 4 years) suffering from myocardial infarction more than 6 months before with or without carotid atherosclerotic lesions. After evaluation by B-mode duplex scanning system of extracranial carotid arteries, the patients were subdivided into two groups: Group 1 (n = 16, with carotid plaques or intima-media thickening) and Group 2 (n = 19, without carotid plaques or intima-media thickening). Eighteen age- and sex-matched subjects were recruited as controls (Group 3). Groups 1 and 2 displayed significantly higher levels of total cholesterol and apolipoprotein B and significantly lower levels of HDL cholesterol and apolipoprotein A1 than Group 3, while serum triglyceride and lipoprotein (a)-Lp (a) levels were significantly higher in Group 1 as compared to the control group. Moreover, Group 1 and 2 displayed significantly higher levels of factor VII, fibrinogen, F1+2, thrombin-antithrombin complex and plasminogen activator inhibitor (PAI) post venous occlusion and significantly lower levels of tissue plasminogen activator (t-PA) post venous occlusion than Group 3. Significantly higher levels of t-PA and PAI pre venous occlusion and significantly lower levels of antithrombin III, C-protein and S-protein were observed in Group 1 as compared to controls. In patients with highest Lp(a) level, the lowest t-PA level post venous occlusion and the highest PAI level post venous occlusion were observed. Our data show an activation of coagulation and a deficient fibrinolysis in survivors of myocardial infarction, particularly in those with associated carotid atherosclerotic lesions. We speculate that this thrombophilic state may play a key role in the pathogenesis of atherosclerotic vascular disease and thromboembolic complications. PMID- 8706062 TI - Early postoperative myocardial ischemia after coronary artery bypass grafting. Detrimental effects of nitroglycerine infusion. AB - This study was aimed at assessing the incidence and mechanisms of myocardial ischemia early after coronary artery bypass grafting and the effects of treatment with nitroglycerine. The electrocardiogram of 35 patients (29 males and 6 females, mean age 61 +/- 8 years) with stable angina and multivessel coronary disease, was monitored continuously for 24 hours after uncomplicated coronary artery bypass grafting. Patients were randomized to receive nitroglycerin infusion or placebo. Fourteen of the 35 patients (40%) had 24 transient ischemic episodes (mean duration 11.8 +/- 3.5 min; range 6-20 min with ST segment elevation in 6, ST segment depression in 7 and both ST segment elevation and depression in 1. Seventy-five per cent of the ischemic episodes occurred within the first 6 postoperative hours. The mean ejection fraction prior to surgery and the mean number of stenosed vessels and of the implanted grafts were similar in patients with and without postoperative ischemia (57 +/- 5 vs 57 +/- 6%, p = 0.86; 2.7 +/- 0.5 vs 2.8 +/- 0.4, p = 0.52 and 3.0 +/- 0.9 vs 3.2 +/- 0.7, p = 0.51, respectively) as well as total bypass time and cross-clamp time (123 +/- 38 vs 124 +/- 18 min, p = 0.89 and 67 +/- 20 vs 70 +/- 14 min, p = 0.68, respectively). The values of heart rate and systolic blood pressure at the onset of the ischemic episodes were similar to those recorded 15 min before (103 +/- 16 vs 106 +/- 18 b/min, p = 0.36 and 119 +/- 12 vs 121 +/- 14 mmHg, p = 0.48). Ischemic episodes were recorded in 9 of the 16 patients (56%) randomized to receive nitroglycerine and in 5 only of the 19 patients (26%) randomized to receive placebo (p = 0.05). Thus, transient ischemic episodes occurring early after coronary artery bypass grafting are not preceded by an increase in myocardial oxygen consumption; they appear to be due, therefore, to a primary reduction in coronary blood flow. Treatment with nitroglycerine is associated with a higher prevalence of ischemic episodes, thus suggesting that myocardial ischemia is unlikely to be caused by spasm of large epicardial vessels or grafts. Myocardial ischemia may be caused, instead, by extracorporeal circulation-induced alterations enhanced by the hypotensive effects of nitroglycerine. PMID- 8706063 TI - Coronary surgery without cardiopulmonary bypass. AB - The need to obviate the risks associated with cardiopulmonary bypass (CPB) in coronary surgery has led to an interest in coronary artery bypass grafting without CPB. From November 1994 to May 1995, 58 patients (49 males and 9 females, mean age 61.8 +/- 9.3 years, range 40-74) were selected for coronary artery bypass grafting without CPB. Three patients had left main stenosis and 6 had left ventricular dysfunction (ejection fraction < 40%). Stable angina was present in 42 patients (27 with low threshold angina) and unstable angina in 16. In 44 patients a routine median sternotomy and in 14 cases a small anterior thoracotomy were performed: in the latter the proximal harvesting of the left internal mammary artery was video-assisted by thoracoscopy. The left internal mammary artery was used in 53 cases; the saphenous vein was used in 36 cases; the radial artery was used in 4 cases; the inferior epigastric artery was used in 2 cases and the right gastroepiploic artery in 1 case. We recorded 1 death (1.7%) and 1 case of postoperative low cardiac output syndrome requiring counterpulsation (1.7%). Perioperative myocardial infarction occurred in 3 cases (5.8%). We did not record noncardiac complications (cerebrovascular, renal failure, prolonged ventilatory support over 24 hours or sternal wound complications). Supraventricular and ventricular arrhythmias were never detected. Mean intensive care unit and hospital stay were 1.1 +/- 0.5 and 5.1 +/- 1.7 days, respectively. In conclusion, according to our experience, "beating heart" coronary surgery is a new promising technique that can be considered alternative in most cases to percutaneous transluminal coronary angio and complementary to conventional coronary surgery. PMID- 8706065 TI - [Rehabilitation of the patient with coronary bypass]. PMID- 8706064 TI - [Circulatory support with Hemopump in cardiogenic shock secondary to papillary muscle rupture]. AB - A 71-year-old woman submitted to multiple coronary artery bypass grafts suddenly developed in the third postoperatory day cardiogenic shock. Transesophageal echocardiography examination and color Doppler showed prolapse of the anterior mitral valve leaflet and detached anterolateral papillary muscle in the left atrial cavity with severe mitral valve regurgitation and increased left ventricular wall kynesis. Maximal inotropic and vasodilator support was not effective and a mechanical circulatory assistance was deemed necessary awaiting for mitral valve replacement not performed on emergency for unavailability of operatory rooms. Hemopump pump-cannula assembly was introduced through a femoral graft and the cannula was advanced in the aorta and positioned in the left ventricle across the aortic valve. Pump rate was set at the maximal speed and as an immediate result, mean arterial pressure increased and mean pulmonary pressure decreased. Global cardiac output during 190 min of assistance was 3.48 l/min at a mean arterial pressure of 81 mmHg. The Hemopump provided 3 l/min of flow with an effective left ventricle unloading. The patient subsequently underwent mitral valve replacement and her postoperative outcome was uneventful and free from complications. PMID- 8706066 TI - [Functional evaluation of prosthetic heart valves with fluoroscopy]. PMID- 8706067 TI - [What does health care mean?]. AB - The important problem of the contemporary health care is the financial and organizational coping with the continuing transformation of the society as a whole. There is no doubt that from the long term point of view, the questions concerning the ethos of health care are essential, are worth asking, and should become a matter of interest for health professionals and the public. PMID- 8706068 TI - [Oncogenes and the malignancy process]. AB - An important group of genes for the development of neoplastic diseases are, in addition to tumour suppressor genes, protooncogenes. The latter are highly preserved genes present in a similar sequence in the cell genomes of different species (yeasts - man). They encode components of biochemical signalling pathways by which external mitotic signals stimulate cell proliferation and products which inhibit cell differentiation. The result of activation of protooncogenes into oncogenes (mutations, chromosomal rearrangements, amplifications, viral insertions, insertion mutagenesis) is in particular hyperstimulation of cells resulting in uncontrolled proliferation. Mutations are of the dominant type, elimination of one allele leads to the transformation of a protooncogene into an oncogene. Oncogenes are classified with regard to the transmission level of the mitogenic signal on which they act. Originally they were detected in the genome of oncogenic viruses. However, they do not form their constant and specific constituent, the virus acts as a vector which transmits cellular protooncogenes (or oncogenes) during the reproductive cycle from one cell to another. The activity of various types of oncogenes is the necessary prerequisite for the genesis and development of various neoplastic diseases. Detection of oncogene alterations provides in some instances important diagnostic, prognostic and therapeutic findings. PMID- 8706069 TI - [Treatment of chronic kidney failure in diabetics with peritoneal dialysis]. AB - The number of dialyzed diabetics in this country is increasing (14.5% of all dialyzed patients in 1993). Haemodialyzation treatment of diabetics has, however, some pitfalls: it is difficult to create a permanent vascular approach, the risk of intraocular haemorrhage during heparinization etc. Therefore some authors prefer peritoneal dialysis in the case of renal failure in diabetics. The technique of insertion of an intraperitoneal catheter and treatment proper are in principle the same as in non-diabetic subjects; the method does not require a vascular approach and heparin is not administered. Other advantages of peritoneal dialysis are the stability of the milieu interieur, the preserved residual renal function and the possibility of intraperitoneal insulin administration which imitates the physiological release from the pancreas. Peritoneal dialysis has, however, also some disadvantages: the main risk is peritonitis. It can hasten atherosclerosis, malnutrition is relatively frequent etc. The main value of peritoneal dialysis is extension of therapeutic possibilities for diabetics with renal failure. PMID- 8706070 TI - [Glycemia increases during treatment of hypertension. 13 years' experience in the treatment of middle-aged men, randomized for treatment with beta blockers and diuretics]. AB - BACKGROUND: Hypertension can no longer be evaluated only with regard to the increase of blood pressure; its clinical picture must comprise also metabolic changes with originate from insulin resistance. METHODS AND RESULTS: The authors submit some incentives from a 13-year long therapeutic follow-up of hypertension in a group of 54 middle-aged men suffering from essential hypertension. From a control group of 38 men they differed as regards blood pressure (174.2 +/- 13.2/112.9 +/- 7.0 mm Hg vs. 126.4 +/- 13.9/77.6 +/- 7.4 mm Hg, p < 0.001), body build (BMI = 28.0 +/- 3.4 kg/m2 vs 23.9 +/- 2.5 kg/m2, p < 0.001), waist/hip ratio 0.98 +/- 0.04 vs 0.94 +/- 0.04, p < 0.05, as well as uric acid serum levels (413.7 +/- 111.0 mumol vs. 362.6 +/- 65.9 mumol/l, p < 0.05) and HDL cholesterol (1.36 +/- 0.30 mmol/l vs. 1.51 +/- 0.48 mmol/l, p < 0.05). However, the fasting blood sugar levels did not differ (5.2 +/- 0.7 mmol/l vs 5.1 +/- 0.6 mmol/l). Although the index of insulin resistance was elevated (6.6 +/- 2.1 vs. 2.84 +/- 1.6, p < 0.01). The blood sugar level rose steadily and reached a level of 18.7% +/- 14.8% (p < 0.01). The increments of the blood sugar level correlated with changes in body weight (1.2% +/- 6.1% r = 0.535, p < 0.001), TAG (4.0% +/- 5.6% r = 0.332, p < 0.05) and the fasting baseline blood sugar levels (r = 0.551, p < 0.001). The mentioned variables contributed to the prediction of increments of the blood sugar level in multiple regression (determined coefficient R2 = 0.51). No relationship with the type of treatment was revealed. 11% of the patients developed during the investigation period symptoms of type 2 diabetes. The development of diabetes could be predicted on the basis of three baseline variables- the BMI, the index of insulin resistance and the basal blood sugar level (logistic regression, r = 0.790. CONCLUSIONS: Hypertension is a constituent of the metabolic syndrome, although the rise of the blood sugar does not directly correlate with manifestation of type 2 diabetes, it is a warning that insulin resistance can influence the effectiveness of provisions of the therapeutic regimen, and last not least, also the development of type 2 diabetes. PMID- 8706072 TI - [The effect of fish oil on metabolic parameters in patients with type 2 diabetes mellitus associated with dyslipidemia]. AB - BACKGROUND: The authors discuss the effect of administration of fish oils rich in n-3 fatty acids in diabetic patients type 2 and draw attention to the possible deterioration of glucose homeostasis. The objective of the investigation was to assess changes of the lipid and carbohydrate metabolism in type 2 diabetics with associated dyslipidaemia after enrichment of the diet with n-3 fatty acids. METHODS AND RESULTS: To 17 patients with type 2 diabetes and dyslipidaemia fish oil containing 5.5 g n-3 fatty acids per day was administered. After six weeks a decline of triglycerides was recorded (-47%, P < 0.01), free fatty acids (-27%, P < 0.01) and a rise of HDL2-cholesterol (25%, P < 0.05). The concentration of apo B, apo A-1, LDL- and HDL cholesterol did not change significantly. There were no significant changes of the blood sugar level, glycosylated haemoglobin and fructosamine. The insulin and C-peptide concentration on fasting (and after glucagon stimulation) did not change significantly. With regard to the HDL2 cholesterol and 18:0 fatty acid concentration in serum the group can be divided into responders (with a decline of glycosylated haemoglobin) and non-responders. The two groups have a reverse trend of blood sugar levels and insulinaemia and differ as to the metabolism of 18:1 n-7 acid. CONCLUSIONS: Enrichment of the diet with n-3 fatty acids in diabetics with dyslipidaemia has a favourable effect on the plasma lipid spectrum without causing deterioration of parameters of diabetes compensation. Among the group of patients some can be found where fish oil administration improves also glucose homeostasis. PMID- 8706071 TI - [Long-term treatment of diabetes with transplantation of a pancreatic segment]. AB - BACKGROUND: Successful transplantation of the pancreas is at present the only way how to ensure on a long-term basis an almost physiological regulation of the carbohydrate metabolism in type 1 diabetics. So far it is, however, indicated mainly in patients with already advanced microangiopathy where at the same time also renal transplantation is planned and long-term experience is so far limited. The objective of the submitted paper is to report on the development of metabolic compensation and its impact on the development of microangiopathic changes in type 1 diabetics where the complete function of both grafts persisted more han five years. METHODS AND RESULTS: From a group of 34 combined transplantations of a pancreatic segment with an obliterated duct and a kidney, implemented in 1983 1988 in the Institute of Clinical and Experimental Medicine, a group of nine type 1 diabetics was followed up where the independence on exogenous insulin and haemodialyzation treatment persisted for or still persists for 5-8 years. After annual intervals the blood sugar level was examined, the intravenous glucose to tolerance test, free insulin levels, glycosylated haemoglobin, an ophthalmological and neurological examination was made, incl. the peripheral and autonomous system, and by means of a standard questionnaire the quality of life before and after transplantation was assessed. In all examined subjects normal blood sugar levels were recorded. The fasting insulin levels in transplant recipients were higher than in healthy subjects (22 vs. 10.2 microU/ml, p < 0.01) while in the course of the blood sugar curve corresponding levels were recorded. Glycosylated haemoglobin remained after 5 years quite or almost normal (4.2 7.2%). The coefficient of glucose assimilation after 5 years varied in the range from 0.7 to 1.9% min. Hypoglycaemic states were not recorded. In none of the recipients in the course of the investigation deterioration of the ophthalmological finding was observed and in three patients improvement was recorded. Symptoms of somatic polyneuropathy improved in all patients but signs of vegetative neuropathy remained unchanged. In all recipients psychic, physical and social rehabilitation as well as the general quality of life improved markedly. CONCLUSIONS: Although the group of investigated patients is so far small, the authors provided evidence that combined transplantation of the pancreas and kidney can influence in a very favourable way the quality of life and development of microangiopathic complications. As the success rate of transplantations of the pancreas in increasing and the risk of surgical complications is declining due to improving surgical techniques, the authors conclude that combined transplantation of the pancreas and kidney is at present the optimal therapeutic procedure in type 1 diabetics with chronic renal insufficiency and that indication for transplantation of the pancreas could be moved to earlier stages of diabetes when it would be possible to influence the development of diabetic microangiopathy more favourably. PMID- 8706073 TI - [Permanent cardiac pacing. Present possibilities and perspectives]. AB - Cardiac pacing is reckoned to be one of the most successful branches of medicine in the second half of the 20th century. Indications for single chamber ventricular pacing are getting gradually restricted. This early pacing mode, which was used in 1958 for the first time, has been recently replaced from modern modes: for atrial, dual chamber and rate adaptive ones. Not only these most recent methods prevent from bradycardia but they also meet demands of modern cardiac pacing, i.e. to restore the synchronization of the atriums and the ventricles and to restore the frequency response. So-called physiological pacing improves both the well-being and the survival of patients and it has also extended pacing indications to the non-arrhythmic ones: first and foremost to the cardiomyopathies. It is dual chamber pacing, the artificial biosensors, various kinds of counters and monitoring functions that enhance the energy consumption. Both low-threshold pacing leads and an automatic adjustment of the pulse energy are helpful in extending the longevity of tiny lithium-powered pacemakers of these days. Current dual chamber rate responsive devices represent an important first step towards a fully automatic smart pacemaker of the future. Such an intelligent pulse generator with several sensors will be operating in the DDDR mode checking automatically the pacing threshold and switching its pacing mode according to pathophysiological requirements. PMID- 8706074 TI - [Renal involvement in monoclonal gammopathies]. AB - Renal affections are a frequent and clinically important complication of monoclonal gammapathies (MG), in particular of multiple myeloma (MM). The main pathogenetic factor in its development is most probably, despite the participation of other nephrotoxic factors, the action of Bence-Jones protein on the renal parenchyma. Mechanisms of renal affection differ - paraprotein (pp) interferes with tubular functions, precipitates in the cells of the tubular epithelium or in the tubular lumen, is deposited in renal tissue in the form of deposits. The main clinical manifestations include proteinuria, impaired tubular functions and reduced filtration capacity of the kidneys. In 5-10% patients renal failure develops, formerly associated with the unequivocally adverse prognosis of the disease. The change of the therapeutic approach which combines haematological and symptomatic treatment, incl. methods of extracorporeal treatment of the blood, however leads in half the patients to improvement or complete restitution of renal functions. The development of the disease is therefore to a considerable extent determined by the activity of the basic disease and its response to chemotherapy. PMID- 8706075 TI - [Epidemiologic autopsy in Alzheimer's disease]. AB - BACKGROUND: The prevalence of Alzheimer's disease (AD) in the population older than than 65 yr. has been estimated in industrialized countries at approximately 10% (8.1-12.5%). AD in the Czech republic in the year 1992 in the official statistics (Institute of Health Information and Statistics, UZIS) has been diagnosed in 22 cases of deceased and 175 sick patients. METHODS AND RESULTS: The crude rate of AD diagnosed by the Khachaturyans method among 2,197 autopsies of people aged 65-99 yr. performed at Thomayer University Hospital from 1988-1992 by use of principle of "epidemiologic" autopsy was estimated in this age group at 0.7%/100,000 thousand men/year and 1.4%/100 thousand women/year. Demographically standardized death rate in AD is 285.25/100 thousand in men and 604.24/100 thousand in women. The prevalence of AD in the whole group was 7.46%. The frequency of AD steeply rises with age with the exception of the tenth decade when the increase stops. CONCLUSIONS: AD is therefore one of most frequent diseases in this country too. AD in this republic is probably not being clinically and autoptically adequately diagnosed or its diagnoses do not become part of official statistics. PMID- 8706076 TI - [Evaluation of the adequacy of hemodialysis from the aspect of urea kinetics models]. AB - BACKGROUND: One of the widely used indicators of adequacy of haemodialysis (HD) is assessment of the Kt/V of urea (K-dialysis clearance of urea, t-time of haemodialysis, V-distribution volume of urea). Work accomplished in recent years suggests that when the HD is adequate, the value of Kt/V should be 1.5. The objective of the present work was to assess to what extent the so-called adequate value of Kt/V can be influenced by the dietary protein intake and the frequency of HD sessions. METHOD AND RESULTS: In 15 subjects on regular haemodialysis treatment the kinetics of urea in serum and in the dialyzation fluid was investigated. The protein intake and residual renal function was also recorded. The mean age of the examined subjects was 49 years (28-65 years). For HD treatment capillary dialyzers FOCUS 120H and ALPHA 160H, cuprophan and haemophan membranes with an area of 1.2-1.6 sq.m were used. The period of HD was 4-5 hours. The mean Kt/V value was 1.31 (+/- 0.20) at a normal level of protein catabolism (NPCR): 1.48 (+/- 0.17)g/kg/day. The mean value of the predialysis serum concentration of urea was 28.8 (+/-52)mmol/l. A formula was derived which makes it possible to calculate the adequate value of Kt/V which respects the intensity of the protein metabolism (NPCR which in a stabilized state is identical with the dietary protein intake), body weight (BW), intensity of urea excretion due to residual renal function (U) duration of the dialysis cycle (T is the sum of the dialysis and interdialysis time expressed in days) and the required value of the predialysis serum concentration of urea (CPRE). The relationship between these variables can be expressed by the following formula: [formula: see text] The calculated adequate value of Kt/V in the examined group varied within a wide range from 1.15 to 1.91 (mean value 1.47 +/- 0.25). The assembled results support the idea that an adequate value of Kt/V cannot be expressed by a single figure and that this value must be evaluated on an individual basis with regard to all measurable factors which influence protein metabolism. PMID- 8706077 TI - [Transmission of the electrocardiogram via telephone. Analysis of 10 years' experience]. AB - BACKGROUND: The problem of impaired cardiac rhythm is in recent years in the foreground of the interest of cardiologists. This was influenced not only by the conclusions of the CAST study which changed the therapeutic pattern used for some years but above all the diagnostic pitfalls of impaired rhythm as regards detection and identification. The objective of the present study was to provide evidence of the advantages and usefulness of the method of telephone transmission of the electrocardiogram within the complex of other diagnostic procedures. METHODS AND RESULTS: The group was formed by 251 patients, incl. 186 men and 65 women, average age 51.7 years (range 19-85 years). The transmission of the ECG by phone was implemented by the system TESLAR (VUIS-CVUT) which comprises recording units and a terminal with a graphic outlet. A total of 3,727 electrocardiographic tracings were assembled. During their analysis the arithmetic mean was used as the characteristic of the position of the group or groups on the scale, the error of the mean expressing the ratio of the standard deviation to the number of cases. In nominal variables most frequently percentage expression was used, frequently with a confidence interval obtained from tables. Evidence of the usefulness of the method is the high rate of detected arrhythmias within emergency calls (88.5%), as compared with normal calls (10.6%) where the yield is close to that of Holter monitoring, however, with the advantage of time ad libitum and thus also a higher yield. CONCLUSIONS: The presentation of long-term experience with electrocardiograms by telephone confirms unequivocally the assumption that the method extends existing diagnostic possibilities in short term, symptomatic, but sporadic arrhythmias, that it makes possible dispensarization of risk patients as well as patients with permanent pacemakers and that it improves the tactics of antiarrhythmic treatment. PMID- 8706078 TI - [Donor selection for allogenic bone marrow transplantation from 1991 to 1995. (The Prague Center for Bone Marrow Transplantation)]. AB - BACKGROUND: The prerequisite of successful transplantation of haematopoietic stem cells is to find a suitable, i.e. HLA matched bone marrow donor. In the submitted paper the authors give an account of the strategy of selection of bone marrow donors, as practised by the Prague transplantation group. METHODS AND RESULTS: In 1991 to 1995 (end of first quarter) the authors sought suitable bone marrow donors for 421 patients where an allogenic bone marrow transplantation was indicated. During that period 95 transplantations were performed (22.5% of all indicated cases), incl. 82 from siblings (86.3%) of the implemented allogenic bone marrow transplantations). A HLA matched donor from the wider family circle was selected for 15 donors, transplantation were performed in 6 patients (6.4% of the implemented allogenic bone marrow transplantations). An unrelated bone marrow donor was sought for 41 patients as no donor was found in the wider family. Transplantation was implemented in 7 of these patients (7.3% of the implemented transplantations. CONCLUSIONS: With the increasing experience of our transplantation centre, no doubt, the ratio of other than sibling transplantations of bone marrow will increase. At present it accounts for 13.7%. The task of our HLA group must be selection of donors matched as well as possible with the recipient according to all available tests of tissue compatibility - from the wider family circle as well as from the register of unrelated donors. PMID- 8706079 TI - [Paclitaxel (Taxol)]. AB - The paclitaxel (TAXOL); Bristol-Myers Squibb Company) represents first agent from novel class of antineoplastic drugs--taxanes to enter routine clinical practice. Paclitaxel interferes with microtubular polymerization by promoting abnormal assembly of microtubules and inhibiting their subsequent disassembly. Pharmacokinetics of paclitaxel has been intensively studied. There are indications for nonlinear pharmacokinetics when paclitaxel is administered as a short infusion and at higher doses. Neurotoxicity, mucositis, and leukopenia correlate with some pharmacokinetic parameters. The clinical development of paclitaxel was initially hampered by hypersensitivity reactions. Current dosage regiments with premedication reduced the incidence of these events to 3%. The major dose-limiting adverse effect of paclitaxel is neutropenia. Significant activities were reported especially in patients with advanced ovarian, breast, non-small cell lung cancer (NSCLC), head and neck cancer and in other types of tumours. Long-term follow-up will also allow the effects of the drug on patient survival to be determined. At present combination of Taxol (paclitaxel) with cisplatin clearly improves the duration of progression-free survival and of overall survival compared with cyclophosphamide and cisplatin in women ovarian cancer. Recently was TAXOL (paclitaxel) registered in Czech republic for treatment of patients with advanced metastatic ovarian carcinoma and in patients with metastatic breast cancer after failure of the standard therapy. PMID- 8706080 TI - [Medical research projects of the Internal Grant Agency in 1995 and 1996]. PMID- 8706081 TI - [Homocysteine, a less well-known risk factor in cardiac and vascular diseases]. AB - Hyperhomocyst(e)mia (Hcy) negatively influences vascular endothelium and coagulation factors. Association of Hcy with premature arteriosclerosis (rather than atherosclerosis), stroke, myocardial infarction and peripheral arterial and venous disease was proved in clinical and epidemiological studies, even as the association with conventional risk factors like age, male sex, smoking, hypertension and hypercholesterolemia. Vitamin substitution of folates, vitamin B6 and B12 decreases Hcy blood levels, however definite evidence is still lacking, whether it results in lower incidence and mortality from cardiovascular diseases. Therefore clinical and epidemiological studies are necessary. Before the grant-application we proved in a pilot study significantly higher Hcy levels in 97 patients with manifest ischaemic heart disease than in 37 controls. PMID- 8706082 TI - [Hyperhomocysteinemia]. AB - Similarly as in other inborn metabolic diseases the cause of hyperhomocysteinaemia are interactions between genetically conditioned changes most frequently due to reduced cystathionine-beta synthase activities and negative factors of the external environment. Negative environmental factors include above all a high dietary animal protein consumption which is the main methionine donor and a low intake of protein of plant origin. Another negative factor is a low intake of foods of plant origin. Fruits and vegetables are among others important sources of folic acid and pyridoxine. Substitution therapy with vitamin preparations is essential in homozygotes and in high risk heterozygotes of cystathionine beta-synthase. This treatment is also necessary during the periconception period in hyperhomocysteinaemic fertile women to reduce the risk of neurotubal defects in their future children. So far investigations are lacking which would provide evidence of a reduced risk of ischaemic heart disease and other cardiovascular diseases in isolated treatment of mildly elevated levels of plasma homocysteine. To elucidate the part played by hyperhomocysteinaemia in hastening of the atherogenetic process further studies are essential, focused on the interaction of elevated homocysteine plasma levels, dyslipoproteinaemias, hyperfibrinogenaemia and other metabolic indicators in this process. PMID- 8706083 TI - [Incidence of sexual abuse]. AB - BACKGROUND: Much attention is paid lately at all levels to sexual child abuse. Its incidence in the population is, however, not known. The objective of the present investigation was to assess the incidence of sexual abuse in a defined group of young people. METHODS AND RESULTS: The authors examined, using an anonymously completed semistructured questionnaire, 311 5th year medical students from Charles University, mean age cca 23 years. They found that 66% of the women and 20% of the men some time during their life were subjected to involuntary sexual interaction. Only one third of the delicts, however, had occurred before the age of 12, the majority after the age of 15 years. The offender was most frequently an unknown man who exhibited before the victim or fingered her. Brutal of serious interactions (rape or attempted rape) were recorded only in 6%. The authors did not record a single case of incest. The majority of examined subjects experienced this event as unpleasant, however, usually did not tell it to anybody. Police organs were informed only in four cases. CONCLUSIONS: The presented negative experience can contribute to the development of some psychiatric disorders and psychosomatic diseases, although in the examined group a higher rate of psychic and sexual complaints was not recorded in probands with a negative sexual experience. PMID- 8706084 TI - [Fibronectin, interstitial collagens and type IV collagen in dermatitis herpetiformis]. AB - BACKGROUND: In the pathogenesis of dermatitis herpetiformis the participation of some proteins of the extracellular matrix has not been elucidated so far. They are found more amply in the papillary derma where they are manifested most markedly as the mentioned type of dermatosis. The objective of the present work was to assess the participation of components of the extracellular matrix in the pathogenesis of diseases and the possibility of their detection to make a more accurate diagnosis. METHODS AND RESULTS: In a group of 11 patients (mean age 39 +/- 17 years) with dermatitis herpetiformis the diagnosis was established on the basis of the clinical and histological finding. Fibronectin and collagens type I, III, IV, V were assessed in the skin using rabbit antisera (INPHARM Co. Moscow). An elevated level of collagen type III and V was found in the affected papillae and on the floor of the blisters in the papillary derma, collagen type IV on the vault of the blister and fibronectin deposits in the affected papillae which had a netlike structure. CONCLUSIONS: Based on the above findings assessment of fibronectin, collagens type III, IV and V in the affected skin can be considered an important supplementary examination in the histological diagnosis of dermatitis herpetiformis. PMID- 8706085 TI - [The CT image of atrophy of the head of the caudate nucleus in Huntington's chorea]. AB - BACKGROUND: The aim of the study was to measure, using computed tomography (CT), the head of caudate nucleus in Huntington's disease (HD) and in controls in order to verify, whether differences in results have a diagnostic value. METHODS AND RESULTS: We measured using CT (orbitomeatal line, 5 mm slices), the head of caudate nucleus are a (HCNA), bifrontal distance (BFD: maximum distance between frontal horns of lateral ventricles), bicaudate distance (BCD: minimum distance between heads of caudate nuclei) and calculated an index (BCD/BFD) in 12 HD patients (mean age 44.6 yrs (SD: 8.8), mean duration of HD 3.6 yrs (2.5), and in 18 age matched healthy controls. We found in HD patients mean HCNA 0.40 cm2 (SD: 0.12) (min. 0.14 cm2, max. 0.55 cm2), in controls 1.20 (0.1) (1.06, 1.45). Mean BFD in HD patients was 3.99 cm (0.37), (3.31, 4.48) and in controls 3.55 (0.45) (2.84, 4.76). Mean BCD in HD patients was 2.74 cm (0.44) (2.06, 3.44) and in controls 1.55 (0.50) (0.79, 2.70) and mean BCD/BFD index in HD patients 0.68 (0.06) (0.59,0.77), and in controls 0.43 (0.10) (0.25, 0.59). The group statistics revealed the significant difference between HD patients and controls: HCNA (p < 0.0001), BFD (p < 0.01), BCD (p < 0.0001) and BCD/BFD index (p < 0.0001). When dealing with individual values, only HCNA shows no overlapping of values between HD patients and controls. CONCLUSIONS: CT linear measurement of HCNA is a simple and specific tool for the diagnosis of HD. PMID- 8706086 TI - [Diet and diabetes mellitus]. AB - Diet is the basic treatment of every diabetic patient. The first diabetic diet was described in detail by John Rollo in 1797. During the second half of the 19th century and first half of the 20th century great changes as regards the composition of the diabetic diet occurred. The findings on the association of atherosclerosis and fat intake led to a reduction of fat in the diabetic diet. A great deal of research in the seventies and eighties of this century was devoted to exchange of foods in the diabetic diet with regard to the carbohydrate content or the rise of the blood sugar after their intake. The possibility to include sucrose and simple sugars in the diabetic diet was also repeatedly tested. Findings on the blood sugar level after ingestion of different meals, but also lipid levels and epidemiological results led to recommendations of scientific associations worldwide, incl. the Czech Diabetological Society. PMID- 8706087 TI - [History of Fodermayer's Pavilion]. PMID- 8706088 TI - [Systemic enzyme therapy. Problems in absorption of enzyme macromolecules]. PMID- 8706089 TI - [Study on geographical distribution of leptospirosis in China]. AB - Since 1955, Leptospirosis has been recognized as a notifiable infectious disease in China. According to the data gathered from 26 provinces of China (except for Taiwan) in 1955-1993, the average morbidity and mortality were 7.08/10(5) and 1.02% respectively. Distribution of the disease involved 26 provinces in China (not including Taiwan). The trend of the disease has been steady for recent years but outbreaks occasionally occurred, especially in several southern provinces of China. Potential risk factors may exist in these areas. Distribution of endemic areas of this disease is located between 25 degrees-35 degrees latitude and 100 degrees-120 degrees longitude i, e. those provinces along with drainage areas of the Yangtze River and the Huaihe River. PMID- 8706090 TI - [A study on the level of antibody against measles through maternal-fetal transfer and the immuno-response to measles vaccine among 4 to 7 month olds]. AB - A total number of 143 infants at the age of 4 to 7 months from Yantai city Shandong was selected for the study of antibody level against measles by maternal fetal transfer and immuno-response to measles vaccine in October of 1993. The results showed that the negative rates of maternal-fetal antibody among infants of 4, 5, 6 and 7 month olds were 75.00%, 81.25%, 94.87% and 90.1%, respectively. The positive rates and geometric mean titers (GMTs) for immuno-response to measles vaccine were 92.86%, 84.38%, 97.44%, 100.00% and 55.17, 42.41, 69.95, 71.46, respectively. There were significant lower immune response to measles vaccine in infants who had high titer ( > 1:2) than those who had low titer ( 0.13), and therefore the interaction of the former was not affected by the effects of the latter as yet. PMID- 8706100 TI - [Relationship between Helicobacter pylori infection other factors and diffuse-and intestinal type stomach carcinomas]. AB - To investigate the role of Helicobacter Pylori (HP) infection and other factors in the development of stomach carcinoma, a matched case-control study was conducted in Dalian, China. A total number of 95 paired cases and controls were involved in the study. The cases were divided into two histological types: diffuse- and intestinal-type. IgG antibody of HP in the subjects was tested with ELISA method. The results revealed that the prevalences of HP infection in cases and in controls were 77.5% and 50.0%, respectively, and that the matched OR was 5.0 (2.08-12.01). The stratified analysis showed that the risk effect of HP in intestinal-type was more significant (OR = 5.75, 1.99-16.63) than it in defuse type (OR = 3.5, 0.73-16.89). Cancer history within family and blood group A were associated with diffuse-type while the environmental factors such as low economic status, irregular eating and fast-eating habits were mainly associated with intestinal-type. We conclude from our findings that HP infection may play a role in the development of stomach cancer, especially for intestinal-type gastric cancer. The two Lauren's histological types of stomach cancer might be different in terms of pathogenesis and characteristics. PMID- 8706102 TI - [Five methods for the estimation of confidence intervals on survival rate]. AB - Descriptive analysis on medical follow-up data often involves estimation of survival rates, such as Kaplan-Meier product limit estimator or life table survival rate for small or large scale data sets, respectively. Five methods for estimating the confidence intervals (CI) of survival rates were described. These methods for estimation of intervals were the classic (based on Greenwood's formula), correctness, arcsine transformation, log (-log) transformation and logit transformation. Two examples were described in detailed for the calculation of 95% confidence intervals. PMID- 8706101 TI - [Application of the polymerase chain reaction to the epidemiological study of amoebiasis]. AB - The polymerase chain reaction (PCR) has been used in studying the pathogenicity of Entamoeba histolytica in a rural community in Shandong province. Formalin fixed stool samples were used for extraction of DNA. The PCR amplifications were performed using two sets of primer that discriminate between pathogenic and nonpathogenic E. histolytica. All of the 32 cases of cyst carrier diagnosed microscopically were identified as positive. Three groups were defined through PCR analysis: 6 cases (18.8%) were positive for pathogenic but negative for nonpathogenic, 25 cases (78.1%) were negative for pathogenic but positive for nonpathogenic, and only one case (3.1%) was positive for both pathogenic and nonpathogenic. The cases found to be pathogenic by PCR were also closely correlated with their clinical manifestations (diarrhea) (OR = 31.5, P < 0.005). All of the control cases showed negative reaction to PCR. PCR has also been used to study the epidemiology of amoebiasis in a small population group with 40 persons in the same province. Both positive rates by PCR on pathogenic and nonpathogenic were 10%. The results showed that this new technique can be applied in the field for the epidemiological studies of pathogenic and nonpathogenic E. histolytica. PMID- 8706103 TI - [Epidemiology and molecular epidemiology of type I diabetes mellitus]. PMID- 8706104 TI - Common heredity of familial hemiplegic migraine and basilar artery migraine? PMID- 8706105 TI - International comparisons in migraine epidemiology. PMID- 8706106 TI - Power spectrum analysis of heart rate fluctuation in cluster headache. PMID- 8706107 TI - Guidelines for controlled trials of drugs in cluster headache. PMID- 8706108 TI - Cervicogenic headache: a personal view. AB - Cervicogenic headache describes pain referred along the upper three cervical roots and is a common source of headache in the over fifties. A review of clinical features, anatomy, and investigations using neural blockade shows several clinical pictures. Diseases of the cranio-vertebral junction, e.g. syringomyelia, and tonsilar descent, can produce mechanical or hydrodynamic head and neck pain. Pre-existent migraine and tension headache may be precipitated or aggravated by cervical spondylosis, but the anatomical mechanism is unclear. Headaches which arise from and are primarily caused by disorders in the cervical spine are exemplified by spondylosis and acute whiplash injury, but are usually transient. Evidence for a cervicogenic mechanism of chronic whiplash headache is, however, lacking. PMID- 8706109 TI - Influence of a selective 5HT1-receptor agonist GR43175 on platelet responsiveness. AB - The possible interaction of sumatriptan, a selective 5HT1-receptor agonist, with platelet responsiveness has been investigated. Stimulation of platelet rich plasma with sumatriptan (1-100 microM) did not induce shape change, aggregation or modification of intraplatelet cytosolic calcium levels. Total inhibition of aggregation induced by 20 microM 5HT was observed in platelets preincubated for 20 min with 100 microM sumatriptan. In the same model, platelet stimulation with 4 microM adenosine 5'-diphosphate (ADP), concentration known to induce an irreversible single-phase curve, determined a decrease of aggregatory response. Concentrations from 1 microM to 50 microM of sumatriptan did not influence the aggregatory response induced by 5HT and ADP. These effects appear not to be determined by modifications of platelet calcium homeostasis. The possibility to modulate platelet responsiveness by sumatriptan offers a further approach for evaluating the probable link between platelet behaviour and pathophysiology of migraine. PMID- 8706110 TI - Is familial hemiplegic migraine a hereditary form of basilar migraine? AB - We studied aura symptoms in 83 patients from 6 unrelated families suffering from familial hemiplegic migraine. Fifty-five of the patients reported symptoms that allowed us to categorize them as basilar migraine (BM) patients, in accordance with the International Headache Society (IHS) criteria. In a control group of 33 patients suffering from migraine with aura and 33 patients suffering from migraine without aura, 9 patients complained of vertigo, and only one patient of diplopia during one of her attacks. None of these control patients fulfilled the IHS criteria for BM. We suggest that familial hemiplegic migraine and BM may share certain pathophysiologic mechanisms, which may consist of a (genetically determined) disturbance of basilar artery blood flow. PMID- 8706111 TI - Migraine, chronic tension-type headache, and cluster headache in an Ethiopian rural community. AB - Fifteen-thousand-five-hundred adults (> or = 20 years) in a rural district in Ethiopia with a population of 250,000 were studied for chronic headache. Door-to door survey was performed by trained lay health workers using a questionnaire with a high degree or reliability and validity. The 1-year prevalence of migraine headache was 3.0% (4.2% for females and 1.7% for males) with the peak age specific rate in the fourth decade. Migraine headache was about three times more common in females than in males at any decade. Two-thirds of migraine sufferers had rather frequent attacks. The most frequent trigger factors were emotional stress (90%), changes of weather (78%), physical exhaustion (75%), and smell (70%). Migraine with aura was rare. Family occurrence of migraine in first-degree relatives was 30%. The 1-year prevalence of chronic tension-type headache was 1.7%, while cluster headache was found to be extremely rare. Compared to similarly performed surveys, the prevalence of migraine among rural Ethiopians was less than among Nigerian Africans. PMID- 8706112 TI - Cerebrospinal fluid analyses in migraine patients and controls. AB - To investigate the role of central neurotransmitters in the pathogenesis of migraine, we measured cerebrospinal fluid (CSF) levels of certain amino acids (glycine, taurine, glutamine) and metabolites of biogenic amines (5 hydroxyindoleacetic acid and homovanillic acid) in 38 migraine patients and compared them with the levels from 10 headache-free controls. The levels of taurine, glycine and glutamine were significantly higher in the migraine patients (p < 0.0001 for taurine and glycine; p < 0.0009 for glutamine); there were no significant differences among the three migraine subgroups (infrequent migraine, frequent migraine and transformed migraine). In seven patients subsequently treated with divalproex sodium, CSF taurine levels decreased significantly from pretreatment baseline values. These data support the concept that migraine is at least in part a disorder of central neurotransmission. PMID- 8706113 TI - Increased cerebral blood flow velocity induced by cold pressor test in migraine: a possible basis for pathogenesis? AB - Noradrenergic nuclei of the locus coeruleus are believed to be involved in migraine pathogenesis. We recently demonstrated a typical intracerebral vasoconstriction after prolonged (5 min) exposure to cold pressor test (CPT), likely related to a central noradrenergic mechanism modulated at the locus coeruleus level and eliminated by pretreatment with clonidine. In the present study, we used transcranial Doppler ultrasonography to monitor blood flow velocity (BFV) changes to CPT in the middle cerebral artery (MCA) of migraine with (MA, n = 12) and without (MO, n = 15) aura subjects. CPT induced a significant increase in BFV and a concomitant decrease in the pulsatility index (PI), a pattern which is the opposite of the results obtained with controls. The results were comparable when controls were pretreated with clonidine. The MO patients produced an intermediate pattern between the MA and control subjects. A possible altered modulatory effect of opioids and/or serotonin on noradrenergic nuclei of the brainstem is the possible cause of the observed inverse response in migraine, suggesting intracerebral vasomotor instability in these patients. PMID- 8706114 TI - Effects of increasing the intracranial blood volume in cluster headache patients and controls. AB - Eleven patients with episodic cluster headache in period, five patients out of period and six controls were studied concerning the effects of an increase of the intracranial blood volume by tilting. Common carotid artery (CCA) blood flow was similar in all three groups at baseline and during tilting. CCA diameters were similar at baseline and increased during tilting in all three groups, indicating that tilting caused an increase in the extra- and intracranial blood volume. Unilateral pain or sympathetic dysfunction did not appear during tilting in the patients out of period or in the controls. In four of eight studied patients with cluster headache in period, unilateral miosis and ptosis appeared during tilting. Two of these four patients developed intense unilateral pain, while the other two did not report any pain. Four other patients developed slight unilateral pain but no sympathetic dysfunction during tilting. PMID- 8706115 TI - Sympathetic-parasympathetic activation during spontaneous attacks of cluster headache: evaluation by spectral analysis of heart-rate fluctuations. AB - Twenty-four hour ECG Holter and blood-pressure monitorings were performed in eight patients suffering from cluster headache. Spectral analysis of heart-rate fluctuation was used to assess the autonomic balance under basal conditions, after head-up tilt, and during a spontaneous attack. Normal autonomic balance was found at rest and during sympathetic activation obtained with head-up tilt in the interparoxysmal period. Before the onset of headache, an increase in the low frequency (LF) component of the power spectrum was apparent in all patients. This sign of sympathetic activation was followed by an increase in the high-frequency (HF) component that developed about 2000 beats after the onset of headache and rapidly overcame the LF component until the end of pain. Significant differences were found when comparing the spectral parameters [total spectral values (TP), power of the LF and HF components and LF/HF ratio] obtained before, during and after headache. During the attack, blood pressure increased and heart rate decreased in all subjects. There appears to be a primary activation of both sympathetic and parasympathetic functions in cluster headache attacks. The sympathetic component seems to be involved mostly in the development of the attack, whereas the parasympathetic activation seems to occur, following the onset of the attack, independently of the pain. PMID- 8706116 TI - Cross-correlation between stress, pain, physical activity, and temporalis muscle EMG in tension-type headache. AB - Thirty-six tension-type headache subjects and 36 non-headache matched controls recorded their temporalis muscle electromyographic (EMG) activity and their pain intensity, stress and physical activity levels in a daily diary. Measurements were performed every 30 min for 6 days (EMG 3 days only). A time-lagged cross correlational analysis between pain, stress, physical activity, and EMG shows that the highest correlation coefficient values occurred between pain and stress at the same (r = 0.33) and at the two preceding 0.5 h time points (r = 0.21 and r = 0.26) in the headache group. Virtually no correlation was found between pain, stress, or physical activity with EMG for either group. These data show that temporalis muscle activity levels were not related to the rise and fall of the subjects' pain or stress levels. Conversely, elevated stress appeared to be highly related to pain; it occurs as both an antecedent and simultaneous event with elevated headache pain. PMID- 8706117 TI - Response measures in the acute treatment of migraine. AB - Pain ratings from 268 migraine patients have been used to compare the visual analogue scale (VAS) and a four-point verbal rating scale. All patients completed pain ratings on both scales at the beginning of a migraine attack and 4 h after starting treatment with sumatriptan or placebo. The VAS scores showed large variability within each category on the verbal rating scale. A common way of analysing pain in migraine studies is to classify patient response as a success or failure based on the score on the four-point verbal rating scale. In this study, the statistical power of analysing response either as "success or failure" or by means of the VAS score has been investigated by stochastic simulation. The simulations showed that the two response measures resulted in approximately equal power. PMID- 8706118 TI - Low-dose ibuprofen in self-medication of mild to moderate headache: a comparison with acetylsalicylic acid and placebo. AB - A double-blind, threefold crossover, double-dummy trial was performed, investigating the efficacy of 200 mg ibuprofen compared with 500 mg acetylsalicylic acid and placebo in patients who usually treated their headaches with over-the-counter drugs. Ninety-five patients suffering from mild to moderate migraine or episodic tension-type headache were included. Seventy-seven patients entered the intention-to-treat analysis and 65 completed all three treatments. For the main response criterion, a minimum 50% decrease of headache intensity on a visual analogue scale at 1 h after treatment, ibuprofen was significantly superior to acetylsalicylic acid and placebo. This was true for migraine attacks and tension-type headache episodes. Towards the end of the observation period (150 min), the differences between ibuprofen and acetylsalicylic acid were no longer significant. In conclusion, ibuprofen was at least equivalent to acetylsalicylic acid and superior to placebo. PMID- 8706119 TI - Visual dysfunction in migraine with aura. PMID- 8706120 TI - Size control: the regulation of cell numbers in animal development. PMID- 8706121 TI - DNA helicases: new breeds of translocating motors and molecular pumps. PMID- 8706122 TI - GASPing for life in stationary phase. PMID- 8706123 TI - The alpha-galactosyl epitope: a sugar coating that makes viruses and cells unpalatable. PMID- 8706124 TI - Acid sphingomyelinase-deficient human lymphoblasts and mice are defective in radiation-induced apoptosis. AB - Stress is believed to activate sphingomyelinase to generate ceramide, which serves as a second messenger in initiating the apoptotic response. Conclusive evidence for this paradigm, however, is lacking. In the present study, we used a genetic approach to address this issue directly. We show that lymphoblasts from Niemann-Pick patients, which have an inherited deficiency of acid sphingomyelinase activity, fail to respond to ionizing radiation with ceramide generation and apoptosis. These abnormalities are reversible up on restoration of acid sphingomyelinase activity by retroviral transfer of human acid sphingomyelinase cDNA. Acid sphingomyelinase knockout mice also expressed defects in radiation-induced ceramide generation and apoptosis in vivo. Comparison with p53 knockout mice revealed that acid sphingomyelinase-mediated apoptosis and p53 mediated apoptosis are likely distinct and independent. These genetic models provide definitive evidence for the involvement of acid sphingomyelinase in one form of stress-induced apoptosis. PMID- 8706125 TI - An alternatively spliced C. elegans ced-4 RNA encodes a novel cell death inhibitor. AB - The C. elegans gene ced-4 is essential for programmed cell death. We report that ced-4 encodes two transcripts and that whereas the major transcript can cause programmed cell death, the minor transcript can act oppositely and prevent programmed cell death, thus defining a novel class of cell death inhibitors. That ced-4 has both cell-killing and cell-protective functions is consistent with previous genetic studies. Our results suggest that the dual protective and killer functions of the C. elegans bcl-2-like gene ced-9 are mediated by inhibition of the killer and protective ced-4 functions, respectively. We propose that a balance between opposing ced-4 functions influences the decision of a cell to live or to die by programmed cell death and that both ced-9 and ced-4 protective functions are required to prevent programmed cell death. PMID- 8706126 TI - Myelination in the absence of galactocerebroside and sulfatide: normal structure with abnormal function and regional instability. AB - The vertebrate nervous system is characterized by ensheathment of axons with myelin, a multilamellar membrane greatly enriched in the galactolipid galactocerebroside (GalC) and its sulfated derivative sulfatide. We have generated mice lacking the enzyme UDP-galactose:ceramide galactosyltransferase (CGT), which is required for GalC synthesis. CGT-deficient mice do not synthesize GalC or sulfatide but surprisingly form myelin containing glucocerebroside, a lipid not previously identified in myelin. Microscopic and morphometric analyses revealed myelin of normal ultrastructural appearance, except for slightly thinner sheaths in the ventral region of the spinal cord. Nevertheless, these mice exhibit severe generalized tremoring and mild ataxia, and electrophysiological analysis showed conduction deficits consistent with reduced insulative capacity of the myelin sheath. Moreover, with age, CGT-deficient mice develop progressive hindlimb paralysis and extensive vacuolation of the ventral region of the spinal cord. These results indicate that GalC and sulfatide play important roles in myelin function and stability. PMID- 8706127 TI - The Drosophila smoothened gene encodes a seven-pass membrane protein, a putative receptor for the hedgehog signal. AB - Smoothened (smo) is a segment polarity gene required for correct patterning of every segment in Drosophila. The earliest defect in smo mutant embryos is loss of expression of the Hedgehog-responsive gene wingless between 1 and 2 hr after gastrulation. Since smo mutant embryos cannot respond to exogenous Hedgehog (Hh) but can respond to exogenous Wingless, the smo product functions in Hh signaling. Smo acts downstream of or in parallel to Patched, an antagonist of the Hh signal. The smo gene encodes an integral membrane protein with characteristics of G protein-coupled receptors and shows homology to the Drosophila Frizzled protein. Based on its predicted physical characteristics and on its position in the Hh signaling pathway, we suggest that smo encodes a receptor for the Hh signal. PMID- 8706128 TI - Alpha L beta 2 integrin/LFA-1 binding to ICAM-1 induced by cytohesin-1, a cytoplasmic regulatory molecule. AB - The avidity of integrin adhesion receptors for extracellular ligands is subject to dynamic regulation by intracellular programs that have yet to be elucidated. We describe here a protein, cytohesin-1, which specifically interacts with the intracellular portion of the integrin beta 2 chain (CD18). The molecule shows homology to the yeast SEC7 gene product and bears a pleckstrin homology (PH) domain. Overexpression of either the full-length cytohesin-1 or the SEC7 domain induces beta 2 integrin-dependent binding of Jurkat cells to ICAM-1, whereas expression of the isolated cytohesin-1 PH domain inhibits T cell receptor stimulated adhesion. Similar inhibition is not exhibited by PH domains taken from other proteins, showing that the interaction is specific and that individual PH domains are capable of discriminating between alternative targets. PMID- 8706129 TI - The MAP kinase pathway controls differentiation from double-negative to double positive thymocyte. AB - T cell development is regulated at two major control points where maturation, proliferation, and antigen receptor gene rearrangement are coordinated. Progression through these developmental control points is dependent upon the expression of different forms of the T cell receptor. Here we show that the MAP kinase cascade is a regulator of the differentiation of immature thymocytes from double-negative to double-positive cell, most probably acting as a transducer of pre-T cell receptor signaling. Furthermore, this study demonstrates the use of retrovirus-mediated gene transfer in fetal thymic organ culture in the analysis of thymic development in mutant mice, an alternative to transgenesis by oocyte injection. PMID- 8706130 TI - Coordinated dual cleavages induced by the proteasome regulator PA28 lead to dominant MHC ligands. AB - The eukaryotic 20S proteasome is known to associate with the IFN gamma-inducible regulator PA28. We analyzed the kinetics of product generation by 20S proteasomes with and without PA28. In the absence of PA28, the 20S proteasome rapidly generates peptides that have been cleaved only once, while internal fragments accumulate only slowly. In the presence of PA28, products generated by two flanking cleavages appear immediately as main products while the generation of single-cleavage products is strongly reduced. Kinetic data support a PA28 induced, coordinated double-cleavage mechanism. In particular, degradation of peptides derived from mouse cytomegalovirus pp89 and JAK1 kinase in the presence of PA28 leads to strongly enhanced production of the respective major histocompatibility complex ligands and potential precursors. These results show that PA28 profoundly alters the cleavage mechanism of the proteasome and appears to optimize the generation of dominant T-cell epitopes. PMID- 8706131 TI - SKP1 connects cell cycle regulators to the ubiquitin proteolysis machinery through a novel motif, the F-box. AB - We have identified the yeast and human homologs of the SKP1 gene as a suppressor of cdc4 mutants and as a cyclin F-binding protein. Skp1p indirectly binds cyclin A/Cdk2 through Skp2p, and directly binds Skp2p, cyclin F, and Cdc4p through a novel structural motif called the F-box. SKP1 is required for ubiquitin-mediated proteolysis of Cin2p, Clb5p, and the Cdk inhibitor Sic1p, and provides a link between these molecules and the proteolysis machinery. A large number of proteins contain the F-box motif and are thereby implicated in the ubiquitin pathway. Different skp1 mutants arrest cells in either G1 or G2, suggesting a connection between regulation of proteolysis in different stages of the cycle. PMID- 8706132 TI - Budding yeast SKP1 encodes an evolutionarily conserved kinetochore protein required for cell cycle progression. AB - The budding yeast SKP1 gene, identified as a dosage suppressor of a known kinetochore protein mutant, encodes an intrinsic 22.3 kDa subunit of CBF3, a multiprotein complex that binds centromere DNA in vitro. Temperature-sensitive mutations in SKP1 define two distinct phenotypic classes. skp1-4 mutants arrest predominantly as large budded cells with a G2 DNA content and short mitotic spindle, consistent with a role in kinetochore function. skp1-3 mutants, however, arrest predominantly as multiply budded cells with a G1 DNA content, suggesting an additional role during the G1/S phase. Identification of Skp1p homologs from C. elegans, A. thaliana, and H. sapiens indicates that SKP1 is evolutionarily highly conserved. Skp1p therefore represents an intrinsic kinetochore protein conserved throughout eukaryotic evolution and may be directly involved in linking kinetochore function with the cell cycle-regulatory machinery. PMID- 8706133 TI - Pathway leading to correctly folded beta-tubulin. AB - We describe the complete beta-tubulin folding pathway. Folding intermediates produced via ATP-dependent interaction with cytosolic chaperonin undergo a sequence of interactions with four proteins (cofactors A, D, E, and C). The postchaperonin steps in the reaction cascade do not depend on ATP or GTP hydrolysis, although GTP plays a structural role in tubulin folding. Cofactors A and D function by capturing and stabilizing beta-tubulin in a quasi-native conformation. Cofactor E binds to the cofactor D-beta-tubulin complex; interaction with cofactor C then causes the release of beta-tubulin polypeptides that are committed to the native state. Sequence analysis identifies yeast homologs of cofactors D (cin1) and E (pac2), characterized by mutations that affect microtubule function. PMID- 8706134 TI - A defect in nurturing in mice lacking the immediate early gene fosB. AB - Although expression of the Fos family of transcription factors is induced by environmental stimuli that trigger adaptive neuronal response, evidence that Fos family members mediate these responses is lacking. To address this issue, mice were generated with an inactivating mutation in the fosB gene. fosB mutant mice are profoundly deficient in their ability to nurture young animals but are normal with respect to other cognitive and sensory functions. The nurturing defect is likely due to the absence of FosB in the preoptic area, a region of the hypothalamus that is critical for nurturing. These observations suggest that a transcription factor controls a complex behavior by regulating a specific neuronal circuit and indicate that nurturing in mammals has a genetic component. PMID- 8706135 TI - Three-dimensional structure of a DNA repair enzyme, 3-methyladenine DNA glycosylase II, from Escherichia coli. AB - The three-dimensional structure of Escherichia coli 3-methyladenine DNA glycosylase II, which removes numerous alkylated bases from DNA, was solved at 2.3 A resolution. The enzyme consists of three domains: one alpha + beta fold domain with a similarity to one-half of the eukaryotic TATA box-binding protein, and two all alpha-helical domains similar to those of Escherichia coli endonuclease III with combined N-glycosylase/abasic lyase activity. Mutagenesis and model-building studies suggest that the active site is located in a cleft between the two helical domains and that the enzyme flips the target base out of the DNA duplex into the active-site cleft. The structure of the active site implies broad substrate specificity and simple N-glycosylase activity. PMID- 8706136 TI - Structural basis for the excision repair of alkylation-damaged DNA. AB - Base-excision DNA repair proteins that target alkylation damage act on a variety of seemingly dissimilar adducts, yet fail to recognize other closely related lesions. The 1.8 A crystal structure of the monofunctional DNA glycosylase AlkA (E. coli 3-methyladenine-DNA glycosylase II) reveals a large hydrophobic cleft unusually rich in aromatic residues. An Asp residue projecting into this cleft is essential for catalysis, and it governs binding specificity for mechanism-based inhibitors. We propose that AlkA recognizes electron-deficient methylated bases through pi-donor/acceptor interactions involving the electron-rich aromatic cleft. Remarkably, AlkA is similar in fold and active site location to the bifunctional glycosylase/lyase endonuclease III, suggesting the two may employ fundamentally related mechanisms for base excision. PMID- 8706137 TI - ATP binding to the sigma 54-dependent activator XylR triggers a protein multimerization cycle catalyzed by UAS DNA. AB - The events that take place at the prokaryotic enhancer of the Pu promoter of Pseudomonas putida prior to the engagement of the sigma 54-RNA polymerase (sigma 54-RNAP) have been studied in vitro. ATP hydrolysis by XylR, the cognate regulator of the system, is preceded by the multimerization of XylR at the enhancer, which is itself triggered by the sole allosteric effect of ATP binding to the protein. Since ADP is unable to support multimerization, ATP hydrolysis might be followed by a return to the nonmultimerized state. This notion is supported further by the properties of mutant proteins that seem to be frozen, in either the nonmultimerized or the multimerized state, respectively. These results support a cyclic mechanism of ATP-dependent association/dissociation of XylR at the promoter UAS that precedes any involvement of the polymerase in transcription initiation. PMID- 8706138 TI - Bioactive saponins and glycosides. II. Senegae Radix. (2): Chemical structures, hypoglycemic activity, and ethanol absorption-inhibitory effect of E senegasaponin c, Z-senegasaponin c, and Z-senegins II, III, and IV. AB - Following the characterization of E-senegasaponins a and b and Z-senegasaponins a and b, new bioactive saponins named E-senegasaponin c and Z-senegasaponin c were isolated from Senegae Radix, the root of Polygala senega L. var. latifolia TORREY et GRAY., together with Z-senegins II, III, and IV. The chemical structures of E and Z-senegasaponins c and Z-senegins II, III, and IV were elucidated on the basis of chemical and physicochemical evidence, and the geometrical isomeric structures of the 4"-methoxycinnamoyl and 3",4"-dimethoxycinnamoyl groups in these saponins were found to show tautomer-like behavior under irradiation with fluorescent lamps. E and Z-Senegasaponins c and E and Z-senegins II, III, and IV were found to exhibit hypoglycemic activity in the oral D-glucose tolerance test. (E) and (Z)-Senegins II also showed an inhibitory effect on alcohol absorption in rats. PMID- 8706139 TI - Synthesis of human C-type natriuretic peptide 22 using chlorotrityl resin and tetrafluoroboric acid deprotection. AB - Human C-type natriuretic peptide 22 (hCNP22), the third member of the natriuretic peptide family, was efficiently synthesized by Fmoc-based peptide chain construction on a 2-chlorotrityl (Clt) resin, followed by deprotection using tetrafluoroboric acid (HBF4). The use of Clt resin was effective in suppressing racemization at the C-terminal cysteine residue caused by the base treatment for Fmoc-cleavage. The disulfide bond of hCNP22 was constructed using the silyl chloride-sulfoxide method to avoid oxidation at the Met residue. Using amino acid and dipeptide-resin derivatives, the effects of bases, protecting groups and resin supports on the racemization at the C-terminal Cys residue were examined in detail. PMID- 8706140 TI - Antirheumatic agents. I. Novel methotrexate derivatives bearing an indoline moiety. AB - Various novel methotrexate (MTX) derivatives bearing an indoline moiety were synthesized and tested for biological activities using human peripheral blood mononuclear cell (hPBMC) and human synovial cells (hSC) derived from patients with rheumatoid arthritis (RA). Compounds having potent activity in vitro were further evaluated using an adjuvant arthritis model in vivo. N-[1-(2,4-Diamino-6 pteridinylmethyl)indoline-5-carbonyl]-L-glutam ic acid 2f showed more potent activities than MTX in vitro and in vivo, and N-[1-(2,4-diamino-6 pteridinylmethyl)-indoline-5-carbonyl]-L-2-ami noadipic acid 2d exhibited fairly good activities in vitro and considerable activity in vivo. Compound 2d was, as expected, not sensitive to folyl-polyglutamate synthetase (FPGS) and did not undergo polyglutamation, a process which may be responsible for a side-effect during MTX therapy. PMID- 8706141 TI - Anticoagulant peptides; synthesis, stability and antithrombin activity of hirudin C-terminal-related peptides and their disulfated analog. AB - We designed a unique anticoagulant decapeptide, which possesses two O-sulfated tyrosine residues, based on the structure of hirudin's C-terminal functional domain. We first prepared a series of octa-, nona- and decapeptides with no sulfation, Suc-Phe-Glu-Pro-Ile-Pro-Glu-Tyr-Tyr-X-OH [X = bond, Leu or Leu-Gln], by a solution phase method and measured their thrombin times (TT) using human thrombin and rabbit plasma. The shortest octapeptide (3a) showed full antithrombin activity comparable to that of the lead compound hirudin (54-65), and the longest decapeptide (3c) prolonged TT most potently with an IC50 value of 5.8 microM. We consequently converted 3c to a disulfated decapeptide (NF-22) with SO3.pyridine complex and compared its antithrombin activity with that of known hirudin-related peptides: hirugen, MDL28050 and hirulog-1. NF-22 showed potent antithrombin activity with an IC50 value of 0.3 microM, being more potent than hirugen and MDL28050 (IC50 values of 4.0 microM and 1.1 microM, respectively). NF 22 was as potent as hirulog-1. NF-22 showed no change in activity in aqueous solution for 10 d at 60 degrees C, and remained about 90% unchanged in rat plasma on incubation for 24h at 37 degrees C, whereas the corresponding unsulfated peptide (3c) was completely digested under the same condition. NF-22 appears to be one of the most potent and stable peptide anticoagulants among the hirudin analogs. PMID- 8706143 TI - Studies on quinolone antibacterials. V. Synthesis and antibacterial activity of chiral 5-amino-7-(4-substituted-3-amino-1-pyrrolidinyl)-6- fluoro-1,4-dihydro-8 methyl-4-oxoquinoline-3-carboxylic acids and derivatives. AB - We previously demonstrated that 5-amino-7-(3-amino-1-pyrrolidinyl) -1-cyclopropyl 1,4-dihydro-8-methyl-4-oxoquinoline-3-carboxylic acid (7) has strong in vitro antibacterial activity even against quinolone-resistant bacteria. We examined optimization of the 3-aminopyrrolidine moiety of 7 by introduction of C-alkyl (Me, Et, Pr, di-Me, cyclopropyl) and N-alkyl groups (Me, di-Me). C-Alkylation at the 4-position of the 3-aminopyrrolidine moiety enhanced in vitro and in vivo antibacterial activity. (S)-5-Amino-7-(7-amino-5-azaspiro[2.4]hept-5-yl)-1 cyclopropyl-pyr rolidinyl) -1-cyclopropyl-6-fluoro-1,4-dihydro-8-methyl-4 oxoquinoline-3-carboxylic acid (15b) showed strong antibacterial activity (in vitro antibacterial activity including quinolone-resistant bacteria is 4 times more potent than that of ciprofloxacin (CPFX) (1); in vivo antibacterial activity is 1.5 to 20 times more potent than that of CPFX (1)) and reduced quinolone toxicity (free from both phototoxicity at a dosage of 30 mg/kg in guinea pigs (i.v.) and convulsion when coadministered with 4-biphenylacetic acid at a dosage of 20 micrograms in rats (i.c.v.)). Their selectivity between DNA topoisomerase II (derived from eukaryotic cells) and DNA gyrase (derived from bacterial cells) was about 3000-fold. PMID- 8706142 TI - Analysis of in vitro iontophoretic skin permeation of sodium benzoate by transport numbers of drug and additives. AB - Effect of species and concentrations of pharmaceutical additives on the iontophoretic transport of benzoate anion through excised hairless rat skin was investigated using a 2-chamber iontophoretic diffusion cell equipped with platinum electrodes at 0 mA for 4h (control) followed by a constant direct current of 0.5 mA for another 4h. One cell facing the stratum corneum was filled with sodium benzoate solution, and the other cell facing the dermis with lithium chloride (LiCl), potassium chloride (KCl) or tetraethylammonium bromide (TEA-Br) solution. Iontophoretic delivery rate of benzoate anion that permeated through skin increased with an increase in the sodium benzoate concentration while maintaining a constant KCl concentration. In contrast, a flux of benzoate anion decreased with an increase in KCl concentration and a constant concentration of sodium benzoate. When KCl was replaced by LiCl or TEA-Br, the flux of benzoate anion was almost the same. These phenomena were evaluated by a concept of transport numbers: theoretical values of benzoate anion flux were very close to the observed data. Potential difference between the skin during the permeation study was also measured between two salt bridges which were connected via calomel electrodes to a potentiometer. It gradually decreased to a certain level in each case, but increased again in some cases. This gradual decrease and increase in the potential difference, in spite of a constant current, were theoretically explained by a gradual increase of ion concentration in the skin membrane and depletion of the cation in the receiver cell, respectively. Analysis of ionic mobility and concentration of penetrants gave a great deal of information on iontophoretic drug permeation through skin. PMID- 8706144 TI - Antitumor-promoting activities of various synthetic 1-O-acyl-3-O-(6'-O-acyl-beta D-galactopyranosyl)-sn-glycerols related to natural product from freshwater cyanobacterium Anabaena flos-aquae f. flos-aquae. AB - 1-O-Acyl-3-O-(6'-O-acyl-beta-D-galactopyranosyl)-sn-glycerol, which was isolated from a nitrogen-fixing fresh water cyanobacterium, Anabaena flos-aquae f. flos aquae, was synthesized by utilizing lipase-catalyzed acylation. The antitumor promoting activities of these galactolipids were evaluated using a short-term in vitro assay of Epstein-Barr virus activation in Raji cells induced by 12-O tetradecanoyl- phorbol 13-acetate (TPA). The glyceroglycolipids which have a palmitoleoyl residue at the 1-O-position exhibited more potent activities than the others in this assay. PMID- 8706145 TI - Synthesis and antitumor activity of phenyl carbocyclic oxetanocin and related compounds. AB - Reaction of trans-2,3-bis[(tert-butyldimethylsilyl)oxymethyl] -1-cyclobutanone (4a) with phenylmagnesium bromide or lithiated aromatic compounds gave two adducts, the (1R, 2R, 3R) isomers (5a--c) and the (1R,2S,3S) isomers (6a--c). The major products (5a, c) were treated with tetrabutylammonium fluoride to give the (1R,2R,3R)-1-aryl-2,3-bis(hydroxymethyl)-1-cyclobutanols (1a, c). The 3-(oxazol-2 yl)-phenyl adduct 5b was converted to the benzamide congener 1b in 6 steps. On catalytic reduction of 1a with Raney Ni the stereochemistry at C-1' was mostly retained, but in the case of 10% Pd-C catalyst, steric inversion occurred. Compounds 1a-c displayed no cytotoxicity towards human nasopharyngeal carcinoma KB cell line. PMID- 8706147 TI - Isolation of pyropheophorbide a from the leaves of Atalantia monophylla (ROXB.) CORR. (Rutaceae) as a possible antiviral active principle against herpes simplex virus type 2. AB - Antiviral activity-guided isolation studies on the leaves of Atalantia monophylla (ROXB.) CORR. (Rutaceae) led to the identification of pyropheophorbide a (1), a simple chlorin derivative, from the chloroform extract (fr. B) as a possible antiviral active principle against herpes simplex virus type 2 (HSV-2). Pyropheophorbide a methyl ester (2) was also isolated from the hexane extract (fr. A). PMID- 8706146 TI - Biological activity of analogues of YM022. Novel (3-amino substituted phenyl)urea derivatives of 1,4-benzodiazepin-2-one as gastrin/cholecystokinin-B receptor antagonists. AB - A series of (3-substituted phenyl)urea analogues of the potent gastrin/cholecystokinin (CCK)-B receptor antagonist YM022 has been prepared. Structure-activity relationship studies of this series suggested that a number of analogues retained good in vitro potency for gastrin/CCK-B receptor. In particular, the (3-amino substituted phenyl)urea derivatives (10-12) were more potent inhibitors of pentagastrin-induced gastric acid secretion in rats than YM022 on intraduodenal (i.d.) administration. PMID- 8706148 TI - [Cloning and sequencing of the gene PE40, coding for a portion of Pseudomonas exotoxin A]. AB - Gene PE40 coding for a mutant form of Pseudomonas Exotoxin A was devoid of its native cell binding domain, and was amplified from the total DNA of P. aeruginosa PA103 by PCR. After being sequenced, it was cloned into expressional vector pBV220 to get a recombinant plasmid, which will be useful in the research of molecular targeted protein. PMID- 8706149 TI - [Changes of collagen distribution in extrapulmonary arteries of hypoxia-induced pulmonary hypertensive rat]. AB - Changes of collagen distribution in extrapulmonary arteries induced by hypoxia and the effects of ligustrazine (ligus) and 764-3 were investigated. Histological study showed that hypoxia could induce an increase of the medial and adventitial thickness of the pulmonary arterial trunk. An increase of type I and type III collagen fibers in pulmonary arterial trunk, particularly the increase of type III collagen fibers in adventitia of the pulmonary trunk were revealed by picrosirius-polarization methods. Ligus partially prevented the increase of type I and III collagen fibers of pulmonary arterial trunk. Electron microscopy showed that both the medial smooth muscle and adventitial fibroblasts were hypertrophic and hyperplastic, and the surrounding collagen fibers were increased. PMID- 8706150 TI - [Study on the effect of endothelin and nitric oxide on DNA synthesis of pulmonary artery smooth muscle cells and modulation of hypoxia]. AB - Increasing vasoconstriction and vascular remodeling of pulmonary vasculature are main characteristics of hypoxic pulmonary hypertension (HPH). Basal pulmonary circulatory tone is maintained by a dynamic balance of vasoconstrictors and vasodilators acting on the pulmonary vascular smooth muscle cells. Endothelin-1 (ET-1) and nitric oxide (NO) are the most important endothelium-derived vasoconstrictor (EDCF) and vasodilator (EDRF). In this report, we studied the effect of ET-1 and NO on hypoxic pulmonary vascular remodeling. We observed that ET-1 dose dependently increased DNA synthesis of pulmonary artery smooth muscle cells (PASMC), SNP, a NO generating drug, inhibited DNA synthesis of PASMC and its inhibitory effect was mediated by cGMP. Hypoxia increased the proliferative response of PASMS to ET-1 by stimulating autocrine of PASMC and decreased the inhibitory effect of SNP by reducing the activity of intra-cellular soluble guanylate cyclase. Based on the result, it is suggested that the balance of ET-1 and NO and regulation of hypoxia play important roles in hypoxic pulmonary vascular remodeling. PMID- 8706151 TI - [The variable number of tandem repeats in the renin gene of rats and possible significance in hypertension]. AB - The variable number of tandem repeats (VNTR) in the first intron of renin gene for the spontaneously hypertensive rat (SHR), its controls Wistar-Kyoto (WKY), renal hypertensive rat, and Sprague-Dawley rat (SD) were compared by polymerase chain reaction (PCR) method. An analysis of VNTR from WKY, Wistar and SD showed that there are two different renin gene alleles and three genetypes 2.0kb/2.0kb, 2.0kb/1.8kb, 1.8kb/1.8kb. The genetype from renal hypertensive rats is same as those seen in the normal controls. However, compared with the WKY, Wistar and SD genes, a "deletion" of approximately 1.0kb was found in the first intron of the SHR renin gene. Our results strongly suggest that the cause and mechanism of elevated blood pressure is complex, and the molecular basis of the genetic-prone hypertension is existed. PMID- 8706152 TI - [High efficiency expression of IL-4 using modified cDNA expression vector in mammalian cells]. AB - In order to express eukaryotic gene in eukaryotic cells, a cDNA library was constructed and screened with synthesized probe of IL-4 cDNA. The obtained cDNA fragment 739 bp in length was inserted into pi H3M plasmid and expressed (8 x 10(3) IU/ml) in COS cells. A pdB-HhIL-4 expression plasmid was constructed using pdKCR and pdBPV-1 plasmid and transfected into mouse C127 cells by calcium phosphate coprecipitation. Transfected cells exhibited altered cell morphology, could be continuously cultured in DMEM essential medium and secreted IL-4 lymphokine into 1% FCS condition culture medium. High biological activity (2 x 10(4)/ml) from the culture supernatant was obtained by FACS analysis of Fc epsilon receptor expression on B cell surface. PMID- 8706153 TI - [Mechanism of medical collagen for soft tissue augmentation]. AB - This paper reports the fate of medical collagen after injected into rats. It is highly purified collagen. The persistence of it in rats is more than one year. It was found that fibroblasts and capillaries grew into the injections. Picrosirius polarization demonstrated that the ration of type I and type III collagen changed. Fine green fibers of type III collagen were predominant at the early stage. Twelve months after implantation, most collagen fibers appeared as thick brilliant red fibers of type I collagen. PMID- 8706154 TI - [Fine stereo structure for natural organic molecules, a preliminary study. II. Melting point influenced by structure factors]. AB - Crystal structures of two compounds from Tripterygium wilfordii Hook f. have been determined by X-ray diffraction method. Structure factors influencing melting point of solid state have been analysed. Crystal class (or space group), recrystallization solvent, force between molecules and fine changes of molecular structures will all cause melting point changes of crystal substance. PMID- 8706155 TI - [Structural changes of extra pulmonary artery and pulmonary arterioles of rats during chronic hypoxia and their significance]. AB - The structural changes of extra pulmonary artery (PA) and pulmonary arterioles and their effects on the pulmonary arterial pressure (PAP) under chronically hypobaric hypoxia (5000 m) were studied. The results showed that (1) The endothelial edema of extra PA and pulmonary arterioles gradually reduced and the endothelial proliferation took place along with the prolongation of hypoxic exposure followed by the recovery of endothelium-dependent relaxation evoked by Ach from depression. This result suggested that the release of E-DRFs from endothelium increased with the structural changes of endothelium in the course of hypoxic exposure, which may be a reason why PAP did not increase any more after 21 to 40 of days exposure. However, the strengthened contraction of PA, the proliferation of cells on the vascular wall, and the muscularity of pulmonary arterioles being related to the hypoxic endothelium may produce a negative effect on decrease of PAP, resulting in persistently higher PAP. (2) The structural changes of pulmonary arteriole endothelium were more severe than those of extra PA endothelium, indicating that pulmonary arterioles might be more sensitive to the stimulation of hypoxia. PMID- 8706156 TI - [Protection from procarbazine induced testicular toxicity by gonadal steroid hormones]. AB - To confirm the claims that the procarbazines (PCZ) effect on stem spermatogonia can be overcome by gonadal steroid hormones,mature,LBNF1 male rats were implanted with silastic capsules containing testosterone (T 200 mm) + estradiol (E 50 mm). The silastic capsules of control and TE groups were removed 24 hours after the PCZ injection and the rats were sacrificed 9 weeks later. The functional status of the testis and the hormonal parameters at the time of the injection of PCZ were measured and correlated with the degree of protection ,as measured by the recovery of sperm counts. The results indicated that testosterone and estradiol does protect spermatogenesis from PCZ intoxification. To determine whether the protection is specific for stem spermatogonia, we also measured body weight loss 3-4 days, lymphocyte counts 1 day, and spermatocyte count 9 days after PCZ treatment. There were no differences in the dose response curves of these three assays between the hormone and sham treated rats. The lack of protection indicates that the hormonal protection of stem spermatogonia was not due to the alterations in either systemic or testicular pharmacokinetics of spermatocyte. We conclude that A3 spermatogonia and preleptotene spermatocyte are sensitive to PCZ and there is no evidence of hormonal protection for the differentiated spermatogonia and preleptotene spermatocyte. PMID- 8706157 TI - [Construction and radioimmunodetection of nude mouse xenograft of human choriocarcinoma]. AB - Fresh tissue of human choriocarcinoma was injected subcutaneously into the BALB/C nude mouse. The xenografts had grown up with 16-68 passages. The hCG level of the ascites or intratumoral blood was 400-20,000 mIU/ml. Radioimmunodetection was performed using 131I labelled murine anti-hCG McAb. The labelled antibody could be specifically accumulated in the tumor sites and the tumor imaging was acquired. The ideal imaging time was 72-96 hours after the injection of antibody. PMID- 8706158 TI - [A primary observation on the effect of cell fusion on metastatic potential of tumor cells]. AB - Mouse spleen cells transfected with pSV2 neo by CaPO4 precipitation were fused with highly metastatic cell clone (PLA801-D95) from human large cell lung cancer cell line. Hybrid cell clone PMS-2 was obtained after G418(400/ml)selection. After injection of 7 x 10(6) PMS-2 cells into nude mice, there was a tumor nodule developed, but the metastatic foci could not be found while 3 x 10(6) PLA801-D95 cells would metastasize to lung and lymph nodes after they were injected into nude mice. It might indicate that sometime mouse spleen cells could not suppress tumor formation but the metastatic potential could be suppressed by the fusion of mouse spleen cells with the lung cancer cells. The results of growth curve, serum independence and incorporation rates of H-thymidine all showed that the growth rates of parental cells were higher than those of PMS-2. Our data suggest that suppression of tumorigenicity and metastatic potential could be controlled by different kinds of genes, and the cloning of metastasis suppressor gene by subtractive hybridization is ongoing in our laboratory. PMID- 8706160 TI - [Establishment of a new culture system for fetal human tracheobronchus]. AB - Human tracheobronchial tissues were cultured in CMRL 1066 medium containing insulin, hydrocortisone, bovine pituitary extract, and trace elements, with or without fetal bovine serum, in a rocked atmosphere chamber. The morphology of the epithelium remained normal for two months after in organ culture, and then, squamous metaplasia appeared in the epithelium and maintained for more than one year. There were continuous epithelial outgrowths from the human tracheobronchial tissues during the whole duration of organ culture, and the epithelial outgrowth can be subcultured in MCDB 151 medium supplemented with growth factors and grew as a tightly packed monolayer of polygonal cells with the presence of keratin and microvilli. The advantages of organ culture and epithelial cell culture were combined in this new culture system. The new culture model has already been used for the investigation of in vitro transformation of human tracheobronchial epithelial cells by chemical carcinogens in our laboratory. PMID- 8706159 TI - [Detection of p53 mutation using PCR-SSCP silver staining method]. AB - A simple and sensitive technique to detect point mutation of the p53 gene using silver staining method for single-strand conformation polymorphism analysis of polymerase chain reaction (PCR-SSCP) is reported. In this study, the aberrations of the p53 gene (exon 6-8) in clinical specimens of primary cervical carcinoma were examined. Of 27 tumor tissues 2 samples showed mutations of p53 exon 7. Absence of abnormal bands in the p53 exon 6-8 in all ten normal cervical tissues. It appears that inactivation of p53 gene by point mutation is infrequent in clinical samples of human cervical carcinomas. PMID- 8706161 TI - [Effect of tripchlorolide (T4) of Tripterygium wilfordii Hook on the production of endothelin-1 by endothelial cells of human umbilical vein]. AB - Tripchlorolide (T4), a single active ingredient of Tripterygium wilfordii Hook was shown to possess both anti-inflammatory and immunosuppressive actions in experimental animals. The paper reports the effect of T4 on the production of endothelin-1 (ET-1) by endothelial cells of human umbilical vein. The cells were incubated with different doses of T4 (2.5, 5, 10 and 20 ng/ml) for 24 hours. By radioimmunoassay the results showed that T4 at concentration of 5 ng/ml significantly inhibited the production of ET-1 by endothelial cell in a dose dependent trend. The significance of T4 action was discussed. PMID- 8706162 TI - [A preliminary observation of EcochG and cochlear morphology after cervical sympathectomy in guinea pigs]. AB - In order to examine the effect of sympathetic nerve on the inner ear, we observed the changes of EcochG and ultrastructure of the hair cells before and after cervical sympathectomy in 13 guinea pigs. We also compared the results with those of the contralateral side (as a normal control group). We found that there were morphologic changes in the stereocilia of the hair cells in the surgical side. No changes were found in the thresholds, amplitudes and latencies of EcochG. It seems that the effect of the morphologic changes are too small to be detected in the electrophysiologic experiments. PMID- 8706163 TI - [Relation between interleukin-1 and periodontal diseases]. PMID- 8706164 TI - [Analysis of HLA-DRB1 alleles in patients with IDDM]. AB - HLA-DR association with IDDM in local population in Beijing was studied by PCR/SSP typing. The frequency of HLA-DR9 was significantly higher in diabetic patients (30.3% [45/148] vs 17.92% [38/212], chi 2 = 6.97, P < 8.3 x 10(-3)). DR3 was higher in diabetic patients in this study (7.0% [10/148] vs 2.36% [5/212], chi 2 = 3.19, P > 0.05) and DR2 was lower in patients with IDDM (7.4% [11/148] vs 19.8% [42/212], chi 2 = 9.67, P < 1.9 x 10(-3)). These results suggest that DR9 and DR3 both were positively associated with IDDM, but DR2 was negatively associated with IDDM. PMID- 8706166 TI - [Transsphenoidal microsurgical removal of GH-secreting pituitary adenoma in: a report of 200 cases]. AB - From July 1979 to April 1992, 212 cases of GH-secreting pituitary adenoma were treated transsphenoidally by microsurgical technique, of whom two hundred cases monitored with their GH levels at preoperative and postoperative periods. The majority of cases totally 132 cases were of macroadenoma, and 68 cases were of microadenoma. 138 cases were operated on after April 1986. The cure and remission rate averaged to 72.5%, rising from 44.4% (before 1986) to 80.5% for microadenoma and from 31.4% to 69.1% for macroadenoma due to improvement of technique and accumulation of experience. One patient died of angiocardiopathy suddenly during operation. Mortality rate was 0.5%. The follow-up observation of postoperative GH levels in 52 cases revealed that the GH levels within 2 weeks after operation were the same as in 3-6 months. It is thought that the early GH levels may represent the surgical results. The surgical technique is described and factors influencing the surgical results, prevention and treatment of operative complications are discussed in this paper. PMID- 8706165 TI - [The cloning and expression of human secreted interleukin 2 receptor alpha chain gene in eukaryotic cells]. AB - By deleting the sequences encoding the cytoplasmic domain, transmembrane domain and the 5' untranslated region, human interleukin-2 receptor alpha cDNA fragment was subcloned into an eukaryotic expression vector pRc/CMV. The recombinant plasmid was then transfected into CHO cells by either electroporation or calcium phosphate precipitation. 11 G418-resistent clones were randomly selected for detection of mRNA expression. 5 clones out of the 11 were proved to be positively hybridyzed with IL-2R alpha cDNA probe. All of the 5 positive cell lines also expressed sIL-2R alpha in their supernatants as detected by ELISA. The establishment of rhsIL-2R alpha cell lines is of importance in the detection and purification of IL-2 based on the ability of affinity binding between IL-2 and its recombinant receptor. PMID- 8706167 TI - [Prediction of retinoblastoma: linkage analysis of families with hereditary retinoblastoma by using polymorphic sites within the Rb gene]. AB - Retinoblastoma (Rb) is one of the most common intraocular tumors in childhood. 40% of the cases are hereditary in nature. 10% are due to the transmission of a germline mutation from an affected parent, and 30% are due to new germ cell mutation. In the hereditary form, the mutation is inherited via a germ cell; therefore, the mutant gene is present in all somatic cells. The segregations of four polymorphic sites within the Rb gene were analyzed in six families with hereditary retinoblastoma. Two families were informed with probe p123M1.8, three were informed with probe p68RS2.0 and one was informed with PCR amplification of XbaI site; all six families were informed with PCR amplification of intragenic VNTR site RB1.20. Our results showed that linkage analysis of families with hereditary retinoblastoma by using intrageneic polymorphic sites could be applied to detect carriers and for prenatal diagnosis in families with hereditary retinoblastoma. PMID- 8706168 TI - [Effect of deoxynupharidine on immune function in vitro]. AB - Deoxynupharidine (DON) is an alkaloid isolated from rhizome Nuphar pumilum which has been extensively used in the treatment of rheumatoid arthritis and back and leg pains as a folk remedy in China. Data presented in this paper indicate that the DON has potent immunosuppressive activities. Mitogens or allogen induced lymphoproliferative responses of murine splenocytes or human tonsillar mononuclear cells (hTMNC) were markedly reduced when DON was added in the cultures. The trypan blue exclusion test showed that this inhibition was not exerted through a toxic effect. IL-2 activity in the supernatants was evaluated for its ability to support IL-2 independant cell line (CTLL-2) proliferation and the results showed that DON had no marked effect on IL-2 level, but inhibited the capacity of murine peritoneal macrophages to produce IL-1 and TNF, which play very important roles in the process of inflammation and immune response. This immunosuppressive actions of DON may partly account for some of its potential in the treatment of chronic inflammatory diseases, where immunological mechanisms are known to play a major pathogenic role. PMID- 8706169 TI - [Anti-promoting and anti-mutagenic actions of G9315]. AB - G9315, a complex extracted from Glycyrrhizae inflata Bat (III), consists of 6 flavonoids with significant antioxidant effects. At 1 mg dose it inhibited the mouse ear edema induced by croton oil, and showed strong anti-promoting effects on two-stage carcinogenesis in mouse skin induced by DMBA plus croton oil. The TPA enhanced 32Pi-incorporation into phospholipid fraction in HeLa cells was inhibited, and the micronuclei in mouse bone marrow cells induced by cytoxan was also depressed. PMID- 8706170 TI - [Effect of RII on the biological properties of human nasopharyngeal carcinoma cell lines]. AB - N(4-hydroxycarbophenyl) retinamide (RII) is a new synthetic analog of retinoids with low toxicity. Its effect on the biological properties associated with tumor malignant behavior of human nasopharyngeal carcinoma cell lines (HNCs), including CNE-2Z parent cell lines and their variants L2, H2, L4 was studied. RII (10(-5) mol/L) caused detectable morphologic differentiation of HNCs. It inhibited growth of HNCs in vitro and decreased ability of these cells to penetrate matrigel coated filters by using a reconstituted basement membrane invasion assay. These results suggested that RII might be a kind of useful anticancer drug. PMID- 8706171 TI - [Detection of trisomy 3 in angioimmunoblastic lymphadenopathy with dysproteinemia by fluorescence in situ hybridization]. AB - Using fluorescence in situ hybridization, we have detected trisomy 3 and the frequency of cells with +3 in 36 patients with AILD. In 36 cases 28 (77.8%) were found having trisomy 3, which was far exceeded that observed in metaphase cytogenetic study (+3 in 15 cases, 42%). Meanwhile, the frequency of cells with +3 was only 4.0%, which was much lower than that of 11.2% in metaphase cytogenetics. The study indicate that the trisomy 3 is a characteristic chromosome aberration of AILD, the cells with +3 constitute only a small part of the specimen, the fluorescence in situ hybridization is more sensitive than metaphase cytogenetics in detection of numerical chromosome aberrations. PMID- 8706172 TI - [Characterization of platelet activation induced by CD9 McAbs]. AB - The platelet activation induced by two anti-human platelet P24/CD9 McAbs was investigated. The results showed that the pathways of platelet aggregation induced by the two McAbs (HI117 and SJ9A4) were different and HI117 and SJ9A4 induced the phosphorylation of platelet proteins (40,000 and 20,000) when platelets were activated. However, HI117 did not cause a rise in intracellular Ca2+ concentration in activated platelets in comparison with SJ9A4 and the epitope recognized by HI117 and SJ9A4 was different and this might explain why the two CD9 McAbs played different roles in platelet activation in addition, McAbs HI117 and SJ9A4 could not promote associations of other proteins (eg: GP II b/III a) with P24/CD9 on activated human platelets. All these results indicate that the mechanisms of platelet activation induced by HI117 or SJ9A4 are different from each other. It suggests that CD9 antigen plays an importent and complex role in platelet activation. PMID- 8706173 TI - [Posterior chamber intraocular lens implantation in the absence of posterior capsular support]. AB - The authors retrospectively studied the clinical records of 15 eyes without posterior capsular support receiving transscleral ciliary sulcus fixation of posterior chamber intraocular lenses. Postoperative followup period range from 2 weeks to 8 months, an average of 2.2 months. Postoperative visual acuity improved in all patients, 20% of which had 0.6 or better and 40%-between 0.2 and 0.5. Lens rotation was the primary complication and accounted for poor acuity in some cases. The results indicate that ciliary sulcus fixation in the absence of posterior capsular support appears to be safe and effective and may represent a significant advance in anterior segment surgery. PMID- 8706174 TI - [A double-blind parallel group study of beclomethasone dipropoinate nasal spray of two different sources in the treatment of allergic rhinitis]. AB - In order to compare the clinical efficacy and safety of beclomethasone dipropionate nasal spray (BDNS) manufactured in Chongqing Glaxo Limited (Group A) and Glaxo UK (Group B), a randomised double-blind parallel group study was performed. A total of 204 patients with seasonal or perennial allergic rhinitis were recruited into the study for a period of 2 weeks. Overall efficacy (excellent/good) was 88% for Group A (excellent 55%) and 90% for Group B (excellent 51%) respectively. Side effects were similar for both groups where dry nose was the most common complaint (4.9% for both groups). However, there was no statistical difference (P > 0.05) between the two groups either in efficacy or side effects. Pollen count was monitored throughout the study period. Pollen season was divided into three periods, i.e. the beginning, the peak and the end. There was no statistical difference (P > 0.05) between the two groups in any period. PMID- 8706175 TI - [A comparative study of fat emulsions clearance test using long chain triglycerides and medium chain triglycerides in volunteers]. AB - Intravenous fat emulsions clearance test using long chain triglycerides (LCT) and medium chain triglycerides (50% MCT/LCT) were studied in ten healthy chinese volunteers. In the morning (8:00 AM) the lipid was infused continuously for 5 hours at rate of 0.17g.kg-1/h. There was an interval of two weeks between LCT and MCT/LCT. The peak concentration of serum triglyceride was smaller in MCT group. The area under concentration curve had significant difference between LCT and MCT groups (6108 +/- 461 vs 4495 +/- 362, P < 0.05). The serum free fatty acid was 628 +/- 42 in LCT and 1049 +/- 1 in MCT respectively in 5 hour (P < 0.001). The concentrations of very low-density lipoprotein (VLDL) in serum were increased in volunteers receiving LCT and MCT, but those of LDL and HDL were decreased. The blood sugar and cholesterol had no significant difference after lipids infusions. The study demonstrated that medium chain triglycerides was faster to be cleared than long chain triglycerides. PMID- 8706176 TI - [Preparation of 125I-F(ab')2-DNR-liposomes and radioimmunoassay of their specific conjugating ability]. AB - The F(ab')2-DNR-liposomes were prepared using F(ab')2 obtained from digested monoclonal antibody against human white cells HI30 (CD45). Their affinity constant for peripheral blood mononuclear cells was determined by radioimmunoassay. Ka = 2.0 x 10(9). A good specific conjugation ability was demonstrated. In preparing the immunoliposomes, it was found that F(ab')2 coupled to the smaller liposomes with higher conjugation ratio. PMID- 8706177 TI - [Study on antifertility mechanisms of tripchlorolide (T4)]. AB - Hyaluronidase activities of testes and epididymides in rats were obviously inhibited by T4 po at dosage of 0.1 mg.kg-1/d for 7 weeks. T4 at dosage of 0.1 mg.kg-1/d for 3 weeks decreased GSH content in epididymides in rats. However, in vitro T4 at concentration of 10 micrograms/ml was found to exert no effect on the content of malondialdehyde in testis or H2O2 in testis and epididymis microsomes in rats. PMID- 8706178 TI - [Photodynamic effect of HPD on DNA functions]. AB - After treatment of the DNA by HPD and light the number of transformants of the M13mp8 DNA obviously decreased during the period of transforming the treated DNA into E coli. Meanwhile some of the mutation sequences appeared in the DNA which recovered from the DNA/HPD/Light transformant. It showed that the wrong replication happened because of the mutated template. Also the in vitro run-off transcription of IL-2R alpha gene was affected by HPD and light. It means that the DNA functions are influenced by the photodynamic effects. PMID- 8706179 TI - [Proliferation of PBMC in patients with Sjogren's syndrome via CD2 pathway]. AB - Primary Sjogren's syndrome is an autoimmune disease whose etiology is unknown. Recent studies show that there is T cell abnormalities in addition to B cell hyperreactivity. In order to better understand the immunoregulatory abnormalities of primary Sjogren's syndrome, cellular immunology has become the main focus of recent studies. As such, the function of CD2 and CD3 pathways constitutes an integral part of these studies. The proliferation of PBMC, non-adhesive cells (mainly T cells) and adhesive cells (mainly B cells) has been investigated in patients with primary Sjogren's syndrome and normal controls; the results show that the proliferation of PMBC and non-adhesive cells in patients is much lower than that in normal controls (P < 0.05), whereas there is no difference in that of adhesive cells between these two groups (P > 0.05). It is also found that the non-adhesive cells' abnormality can not be adjusted by adding adhesive cells of normal controls. In addition, it seems that there is a relationship between the proliferation of PBMC via CD2 pathway in patients and the positivity of anti-SSA and anti-SSB antibodies. However, the underlying mechanism behind the pathogenesis of primary Sjogren's syndrome has yet to be fully understood. This study warrants further research into gaining a better concept of the CD2 pathway at molecular levels. PMID- 8706180 TI - [Primary carcinoma of the middle and distal third of the common bile duct, diagnosis, treatment and results]. AB - The authors present a series of 21 patients with primary carcinoma of the middle and distal third of the extrahepatic biliary tract observed between 1981 and 1994. Indications and limits of the adopted diagnostic protocol and the selection criteria of the patients, identified by evaluating the preoperative resecability, are discussed. The indications for therapy considered the location of the neoplasia and the definitive staging obtained with the intraoperative ultrasonography and the histopathological examination. Seventy six percent of the operations were curative and the mean survival was 40 months range 13-120 mth for patients with tumor of the middle third and of 33 months range 10-133 mth for patients with tumor of the distal third. Survival results in patients who underwent palliative surgery seem to be similar to those obtained with non surgical palliative methods. PMID- 8706181 TI - [Results of endoscopic biliary drainage in primary tumors of the common bile duct (cholangiocarcinoma)]. AB - Endoscopic drainage is an effective therapy for palliation of patients with bile duct carcinoma. From November 1987 to November 1994 112 patients with cholangiocarcinoma were observed. The success rate of the procedure was 94.5%. The biliary drainage was successful in 99% of patients (in 4% of patients the complete biliary drainage was obtained with the aid of a transhepatically inserted guide-wire with the "rendez-vous" procedure). In 15 patients an intraluminal radiation therapy with Iridium-192 was associated. The morbility was 13% and the 30-days mortality 11%. A surgical operation was performed in 26% of patients with radical intent. The mean survival time in patients treated only endoscopically was 208 days for patients with lesions of the distal and middle third of the bile duct and 281 days for patients with hilar cholangiocarcinoma. In patients with intraluminal radiation therapy the mean survival time was 357 days, with a significative difference with the only endoscopically treated group (357 vs 238 days, p < 0.05). We conclude that endoscopic drainage is a safe and effective management for patients with cholangiocarcinoma and it does not preclude definitive surgical treatment in appropriate candidate. PMID- 8706182 TI - Klatskin tumour: radical and palliative treatment. AB - Primitive neoplasms of proximal extrahepatic bile ducts keep their peculiar morphological and spreading patterns, as firstly described by J. Klatskin. Diagnostic and therapeutic approach were modified in the last year, on the basis of technological progress in imaging and more aggressive surgical attitude. Authors reviewed their clinical experience from 1970 to 1995 concerning proximal extrahepatic bile ducts tumors management, mainly evaluating the evolution of diagnostic work-up and the role of resection. Preoperative work-up is now trimed to non invasive techniques, in order to evaluate the intra and extra biliary diffusion; PTC-PTBD performed preoperatively give a clear biliary map, and could be the first step of a palliative definitive treatment in case of non operable patients. Radical resection remains the gold standard of therapy, with the best long-term results. Palliation must be obtained by the easiest comfortable method for the patient (i.e. self-blocking percutaneously inserted endoprosthesis). PMID- 8706183 TI - [Laparoscopic cholecystectomy for gallbladder stones]. AB - The authors describe the technique for the treatment of gallbladder stones using a laparoscopic approach and discuss the diagnostic and operative flow chart stressing complications and ways to avoid them. A total of 2517 non-selected patients underwent surgery since october 1990 up to september 1995. 252 were affected by acute cholecystitis (10%); 172 underwent emergency laparoscopic cholecystectomy. ERCP was performed in 278 patients (11.04%): 177 underwent endoscopic sphincterotomy and laparoscopic cholecystectomy, 21 underwent laparoscopic cholecystectomy before sphincterotomy, 8 laparoscopic cholecystectomy and ESWL. Laparoscopic cholecystectomy was converted into laparotomy in 37 patients (1.4%); surgery was abandoned in 3 patients following to onset of intense bradycardia. Major complications were observed in 0.63%; bile duct injury occurred in four patients (0.15%). One patient died following a massive intraoperative myocardial infarction. Average operative time was 21 minutes. Only 22.8% of patients required mild analgesia on the first day after surgery. The average hospital postoperative stay was 2.6 days. Return to work took place in 98% of non complicated patients within one week of being discharged from hospital. PMID- 8706184 TI - [Cholelithiasis in the patient with cirrhosis of the liver: surgical aspects]. AB - Open cholecystectomy in cyrrotic patients with good liver functions has operative mortality similar to normal subject (0.5-1%), while in patients with severe hepatic cyrrosis mortality varies between 7 and 83%. In this study we have evaluated the post operative results in cyrrotic patients undergone to open cholecistectomy in order to evaluate indications, controindications and risk factors related to surgery. In the last 7 years 34 patients with liver cyrrosis have been operated for biliary calculi, one of them had laparoscopic cholecystectomy. Morbidity was 29.4% (10 cases) mortality 8.8% (3 cases). Jaundice was the main indication for emergency (66.6%) in the two cases it was related to uncompensated liver functions. Endoscopic Retrograde Cholangio Pancreatography (ERCP) can demonstrate today the causes of jaundice avoiding unecessary operations. Post operative evaluations of our series confirm that operative risk is strictly related to epatic disease and to an appropriate surgical option. Moreover patients with compensated liver cyrrosis, (Child A), do not represent anymore a controindication to laparoscopic cholecystectomy that has less septic post operative complications when compared to open surgery. PMID- 8706185 TI - [Congenital dilatation of the common bile duct in an adult]. AB - The Authors present a case of congenital dilatation of the common bile duct in adult life, selecting this lesion from the cystic dilatations of biliary tract group. The type I of biliary tract cystic dilatations is, in fact, considered a clinical-pathological situation apart, that, because of his frequence and his peculiar characteristics especially present in the adult, puts particular problems regarding his arrangement an treatment. The most recent aspects of etiophatogenetic and therapeutic problems of this lesion are treated. The increased risk of neoplastic degeneration in the adult, the recognition of an anomalous pancreaticobiliary duct junction as etiophatogenetic fondamental cause and the necessity of a radical exeretic treatment in in opposition to the old therapheutic orientations are especi-5ally considered. PMID- 8706186 TI - [Peritoneojugular shunt in the treatment of idiopathic ascites in the hemodialyzed patient: a clinical case]. AB - The case of an uremic patient receiving dialytic treatment, who also had an hydiopatic ascites, probably due to peritoneal hyper-waterproofing, is described. Peritoneo-venous shunt was positioned with good result. Generally, similar clinical situations such a therapy is as valid as other commonly used methods. PMID- 8706187 TI - [Surgical treatment of refractory ascites with peritoneovenous shunt]. AB - Leveen and associates described a peritoneo-venous shunt which proved to be useful in patients with intractable ascites. Medical therapy, paracentesis and peritoneovenous shunt have been compared, but there is uncertainty about their relative merits. Therefore, the leveen device has be placed in last years in an increasing number of patients: it has not been shown by prospective trials to prolong survival significantly, although it may shorten hospitalization and improve the quality of life. A number of early and late complications were described but they do not influence the general results: origin and features of such complications are discussed as well as their prevention and treatment and personal cases are presented. Selection of patients and careful surgical procedure seems to be mandatory for better results. PMID- 8706189 TI - Surgical therapy for hilar bile duct cancer. AB - Surgical treatments for hilar bile duct cancerare discussed in relation to the necessity for and most appropriate method of preoperative biliary drainage, desirable operative procedures for achieving long-term survival and the usefulness of portal venous branch embolization before extended hemihepatectomy. We believe that extended right hemihepatectomy can be performed safely after the volume of the remaining liver has been increased by portal embolization and the serum total bilirubin level has dropped to below 2.0 mg/dl. This procedure is a suitable surgical choice for achieving long-term survival in patients with hilar bile duct cancer. PMID- 8706188 TI - [Complications of elective hepatic resections. Analysis of risk factors]. AB - In the last decades the hepatic surgery has been more and more employed thanks to improvement of the surgical technique and of the post-operative assistance which have brought the peroperative mortality of principal specialistic centres to less of 5%. The main post-operative complications which trouble the hepatic surgery, forming in same cases the cause of the death, are: hepatic insufficiency, haemorrhage, subphrenic abscess and the appearance of biliary fistulas. These complications are often connected and linked to the devitalization of a part of the residual parenchyma. We have made a retrospective study on a series of 214 hepatic resections, executed in election, to estimate the main pre and intra operative risk factors. The operative mortality has been zero whereas the post operative one is occurred in 4.2% of the cases with a morbidity of 27.5%. In our experience the meaning full factors to prefigure an operative risk are resulted: the associated pathologies like diabetes, cardiopathies, ipertension and bronchopathies; the length of the operation; the entity of the peroperative haematic loss and of the consequent transfusional therapy and eventually quality the residual parenchyma. PMID- 8706190 TI - Our links with the past: the value of knowing our history. PMID- 8706191 TI - Ondansetron is a better prophylactic antiemetic than droperidol for tonsillectomy in children. AB - Both intravenous ondansetron (OND) and droperidol (DROP) have been observed to reduce vomiting after tonsillectomy in children. This randomized, double-blind investigation compared the effect of OND and DROP on vomiting after outpatient tonsillectomy in 276 healthy children age 2-12 yr. All subjects received a standardized anaesthetic, which consisted of induction with either propofol or halothane/N2O, vecuronium 0.1 mg x kg(-1) on an as needed basis, maintenance with halothane/N2O, midazolam and codeine, and reversal of neuromuscular blockade with neostigmine and atropine on an as needed basis. Subjects were given either OND 150 micrograms x kg(-1) or DROP 50 micrograms x kg(-1)iv after induction of anaesthesia. Rescue antiemetics in the hospital were administered to patients who vomited X 2 and X 4, respectively. Postoperative pain was treated with morphine, codeine and/or acetaminophen. For 24 hr following surgery, emesis was recorded by nursing staff while subjects were in the hospital, and by parents following discharge from hospital. The two groups were similar with respect to demographic data, induction technique and anaesthesia time. The frequency of in-hospital emesis was 16% in the OND-patients and 30% in the DROP-group, P <0.05. The OND subjects required fewer rescue antiemetics, 5% vs 13%, P <0.05. The overall incidence of emesis was 45% in the OND-group and 57% in the DROP-group, P <0.05. In conclusion, ondansetron was a superior prophylactic antiemetic for tonsillectomy in children when compared to droperidol. PMID- 8706192 TI - Prevention of postoperative nausea and vomiting with granisetron: a randomized, double-blind comparison with droperidol. AB - The effects of granisetron for preventing postoperative nausea and vomiting were investigated in a randomized, double-blind comparison with droperidol and placebo in 100 patients undergoing general anaesthesia for major gynaecological surgery. The patients received a single dose of either granisetron (40 micrograms x kg( 1), n = 25), droperidol (1.25 mg, n = 25; 2.5 mg n = 25) or placebo (saline, n = 25)iv over two to five minutes immediately before induction of anaesthesia. The antiemetic effects of these drugs were evaluated during the first three and the next 21 hr after recovery from anaesthesia. During 0-3 hr after anaesthesia, the frequency of nausea and vomiting was 60%, 12%, 16% and 12% after administration of placebo, granisetron, droperiodol 1.25 mg or droperidol 2.5 mg, respectively. The corresponding frequencies during 3-24 hr after anaesthesia were 44%, 8%, 36% and 12%. The efficacy of granisetron in preventing postoperative nausea and vomiting was almost equal to that of droperidol 2.5 mg. The awakening time in the patients who had received droperidol 2.5 was prolonged by approximately three minutes compared with the placebo group (P <0.05), and postoperative drowsiness/sedation was observed in these patients. In conclusion, preoperative prophylactic administration of granisetron is superior to that of droperidol in the prevention of postoperative nausea and vomiting after anaesthesia. PMID- 8706193 TI - [Tonsillectomies and the reinforced laryngeal mask]. AB - One hundred children were randomly allocated to receive general anaesthesia for tonsillectomy either through a performed tracheal tube or a reinforced laryngeal mask. The insertion of both devices was easy but occasionally, with the laryngeal mask, the airway became obstructed during the Boyle-Davis gag insertion. There was more coughing after intubation (ET 26%, RLM 0%, P< 0,001) and extubation (ET 34%, RLM 0%, P< 0,001), than after insertion and withdrawal of the RLM. Oxygen desaturation (ET 14%, RLM 2%, P< 0,002) and internal contamination with blood seepage (ET 30%, RLM 4%, P< 0,001) occurred less frequently than with the RLM. PMID- 8706194 TI - Sublingual triazolam versus peroral diazepam as a premedication for general anaesthesia. AB - Sublingual triazolam 0.2 mg (T) was compared with peroral diazepam 10 mg (D) as a premedicant in a randomised, double-blind study. Eighty-one ASA I-III patients aged 18-70 yr, scheduled for elective surgery and general anaesthesia were studied. The patients were premedicated about one hour preoperatively. The T group subjects (n = 41) received triazolam sl after a placebo po and the D-group subjects (n = 40) diazepam po before a sl placebo. Anxiety and sedation were evaluated before premedication, every 15 min after that until the patient was removed to the operating room, just before the induction of anaesthesia and both 30 and 60 min after operation. Anxiety and sedation were evaluated by the patient using a visual analogue scale (VAS) and by the anaesthetist with a scale of 0-3 for anxiety and 0-4 for sedation. The patients' experience with regards to their premedication and visit to the operating unit were investigated after the operation. In both groups sedation and anxiolysis became different at 30-45 min after premedication, but at the time just before the induction of anaesthesia there was sedation and anxiolysis only in the T-group. There was no difference between the groups at any time. The T-group patients were more satisfied with their premedication and visit to the operating unit. The study drugs did not cause any cardiorespiratory or other side effects. We conclude that triazolam 0.2 mg sl is at least as effective a premedication as diazepam 10 mg po, that is suitable for patients that cannot swallow, and that the patients were more satisfied with it than with diazepam. PMID- 8706195 TI - Attenuation of the catecholamine response to tracheal intubation with oral clonidine in children. AB - We conducted a prospective, randomized, double-blind , controlled clinical trial to examine (1) whether plasma catecholamine (CA) concentrations increased in response to tracheal intubation in children, and (2) the effects of clonidine on the CA responses. Sixty children (ASA physical status I) aged 7-13 yr were allocated to one of three groups (n = 20 for each group): diazepam 0.4 x kg(-1) (active control), clonidine 2 micrograms x kg(-1), or clonidine 4 micrograms x kg(-1) po. These agents were administered 105 min before induction of anaesthesia followed by oral atropine 0.03 mg x kg(-1) given 60 min before anaesthesia which was induced with thiamylal 5 mg x kg(-1) and tracheal intubation was facilitated with vecuronium 0.2 mg x kg(-1). Laryngoscopy, lasting 30 sec, was attempted two minutes after administration of the induction agents. Serial values for blood pressure, heart rate, and venous plasma CA concentrations were compared among the three groups and with the respective preinduction measurements. Children receiving diazepam or clonidine 2 micrograms x kg(-1) showed remarkable increases in systolic and diastolic blood pressures, heart rate, and plasma CA concentrations in response to tracheal intubation (P < 0.05). The increases were similar for the two regimens. These haemodynamic and CA changes were smaller in children receiving clonidine 4 micrograms x kg(-1) (P < 0.005). The haemodynamic responses were positively correlated with the CA responses. These findings indicate that tracheal intubation following rapid sequence induction of anaesthesia in children provokes a reflex increase in sympathetic activity characterized by increased plasma CA concentrations, and that attenuation of the cardiovascular changes with a high oral dose of clonidine may be due to suppression of the increase in sympathetic activity evoked by the intubation. PMID- 8706196 TI - Prophylaxis against the systemic hypotension induced by propofol during rapid sequence intubation. AB - The objective of this study was to determine the effectiveness of two prophylactic approaches against the anticipated hypotension induced by propofol during rapid-sequence intubation. Thirty-six male or female nonpremedicated ASA class I-II patients aged 21-60 yr undergoing elective outpatient surgery were included in the study. Patients were randomly allocated to receive pre-induction ephedrine sulphate (70 micrograms x kg(-1)iv), pre-induction volume loading (12 ml x kg(-1) Ringer's lactate) or no treatment. Rapid-sequence intubation with cricoid pressure was then performed with propofol (2.5 mg. x kg(-1)) and succinylcholine (1.5 mg x kg(-1). The lungs were subsequently ventilated with 0.25-0.5% isoflurane in a 2:1 N2O/O2 mixture. Vecuronium was given once neuromuscular function had recovered from the succinylcholine. Heart rate and systemic arterial blood pressure were measured non-invasively before induction, after propofol administration and every minute for ten minutes after intubation. Pre-induction volume loading prevented the hypotension observed before surgical stimulation in control and ephedrine groups. Moreover, pre-induction volume loading was not associated with increases in heart rate after intubation as was ephedrine administration. The intubating conditions were excellent to satisfactory in most patients and the overall incidence of adverse events during intubation was mainly due to pain during injection of propofol. The present study showed that preoperative volume loading is more efficacious than pre-induction administration of ephedrine sulphate in maintaining haemodynamic stability during rapid-sequence induction with propofol and succinylcholine. In addition, propofol in combination with succinylcholine provides excellent conditions for rapid sequence intubation. PMID- 8706197 TI - Dose-response relationships for edrophonium antagonism of mivacurium-induced neuromuscular block during N2O-enflurane-alfentanil anaesthesia. AB - The purpose of this study was to determine the dose-response relationships for edrophonium antagonism of mivacurium-induced neuromuscular block. Seventy-five ASA I or II adults were given mivarcurium 0.15 mg x kg(-1) followed by an infusion (7 micrograms x kg(-1) x min(-1) during alfentanil-propofol-N2O enflurane anaesthesia. Train-of-four stimulation (TOF) was applied to the ulnar nerve every 20 sec and the response of the adductor policis was recorded (Relaxograph NMT-100, Datex, Helsinki, Finland). Mivacurium infusion was modified at five-minute intervals in order to keep the height of the first twitch in TOF(T1) at 5% of its control value. At the end of surgery, edrophonium (0.0, 0.125, 0.25, 0.5, or 1.0 mg x kg(-1) combined glycopyrrolate (0.0, 0.0012, 0.0025, 0.005, or 0.01 mg x kg(-1) were administered by random allocation. Edrophonium doses of 0.25, 0.5 and 1.0 mg x kg(-1) were different from placebo with regard to time to attain a TOF ratio (fourth twitch in TOF/T1) = 0.7 (13.8 +/- 4.5, 11.1 +/- 3.5, 11.4 +/- 3.0 vs 19.7 +/- 4.7 min P< 0.05). Doses of 0.5 and 1.0 mg x kg(-1) permitted faster recovery time of T1 from 10 to 95% (T10-95) than did placebo (7.5 +/- 3.8, 8.9 +/- 3.5 vs 14.5 +/- 5.0 min P<0.05). Edrophonium 0.5 mg x kg(-1) was different from placebo with regard to recovery time of T1 from 25 to 75% (T25-75) (3.3 +/- 2.0 vs 6.7 +/- 2.0 min P<0.05). Only edrophonium 0.5 mg x kg(-1) provided faster recovery than placebo with regard to all three indices. It is concluded that edrophonium 0.5 + glycopyrrolate 0.005 mg x kg(-1) allow the fastest recovery from a mivacurium-induced block during enflurane-N2O anaesthesia. PMID- 8706198 TI - Clinical comparison of sevoflurane and isoflurane when administered with nitrous oxide for surgical procedures of intermediate duration. AB - The purpose of this study was to compare the haemodynamic effects and emergence times of anaesthesia with sevoflurane with those of isoflurane when the agents were administered with nitrous oxide to adult patients (ASA I and II) undergoing surgery of at least an hour in duration. Fifty patients were randomly assigned to receive either 0.65 minimum alveolar concentration (MAC) (1.3%) sevoflurane or 0.65 MAC (0.8%) isoflurane together with 60% nitrous oxide following induction with thiopentone, fentanyl, and succinylcholine. Systemic blood pressure and heart rate trends were similar for both groups for the duration of anaesthesia. However, differences in systolic blood pressure measurements were noted at one minute after incision (99 +/- 3 mmHg, mean +/- SE, in the sevoflurane group compared with 109 +/- 4 mmHg for isoflurane), and at emergence (125 +/- 3 mmHg for sevoflurane, 134 +/- 3 mmHg for isoflurane), and in diastolic blood pressure measurements at five minutes after intubation (64 +/- 2 mmHg for sevoflurane, 73 +/- 3 mmHg for isoflurane). Recovery of response to command was more rapid after discontinuation of sevoflurane-nitrous oxide (9.9 +/- 1.1 min) than after isoflurane-nitrous oxide (13.9 +/- 1.3 min). Despite earlier emergence, patients who had received sevoflurane did not request postoperative analgesia sooner. We conclude that the purported advantages of sevoflurane, namely haemodynamic stability and rapid emergence, can be expected even when the agent is administered at 0.65 MAC (1.3%) in nitrous oxide to a typical adult surgical population undergoing procedures of intermediate duration (2.3 +/- 0.2 hr). PMID- 8706200 TI - The anaesthetic management of the child with Eisenmenger's syndrome. AB - There is little clinical data in the literature on the anaesthetic management of paediatric patients with Eisenmenger's syndrome undergoing non-cardiac surgery. This paper reviews our experiences with either such patients who underwent a total of 11 surgical procedures. Of the eight children, six had Down's syndrome and an atrio-ventricular septal defect, one had a ventricular septal defect and one an atrial septal defect. Nine of the eleven operations consisted of minor dental, plastic or ENT procedures, while one patient underwent two laparotomies. Premedication (trimeprazine/ meperidine combination or midazolam) was administered on three occasions. Induction of anaesthesia was achieved by either inhalation of halothane (2), or intravenously with thiopentone (6), ketamine (2) or propofol (1). Muscle relaxation and mechanical ventilation were employed only for both intra-abdominal procedures, otherwise patients were allowed to breathe spontaneously with, or without, manual assistance. Halothane (8), isoflurane (2) and enflurane (1) were all used for maintenance of anaesthesia. Non-invasive monitoring was applied intraoperatively for minor procedures, and arterial and central venous catheters inserted for the laparotomies. Postoperative analgesia for both these cases was provided by an epidural infusion of bupivacaine 0.125% and fentanyl 5 micrograms x ml(-1). A single im bolus of morphine was required following a dental clearance, otherwise pain relief for the rest of the cases was achieved by local anaesthetic infiltration and NSAIDS. With the exception of a single episode of bradycardia, induction, maintenance and recovery from anaesthesia were well tolerated in all cases. In conclusion, our experience suggests that despite theoretical risks, children with Eisenmenger's syndrome appear to tolerate a variety of anaesthetic techniques. PMID- 8706201 TI - Non-invasive continuous haemodynamic and PETCO2 monitoring during peroperative cardiac arrest. AB - We describe a cardiac arrest which occurred during general anaesthesia in the prone position for surgical correction of lumbar kyphosis in a patient with Marfan's syndrome. Peroperative monitoring was routine with ECG, non-invasive arterial pressure, oximetry, PETCO2 and central venous pressure, plus aortic blood flow and and systolic time intervals via an oesophageal echo-Doppler device. Forty-five minutes after the start of surgery, a sudden decrease in aortic blood flow followed by a decrease in PETCO2 suggested acute cardiac failure despite continuation of the ECG signal. Initial CPR in the prone position produced a slight increase in PETCO2. When the patient was turned to the supine position and the legs elevated, chest compression was more efficient and spontaneous circulation was rapidly restored. Circulatory arrest could be explained by incompletely treated hypovolaemia, or by myocardial depression (decrease in aortic blood flow and lengthened pre-ejection period) combined with excessive hypotension in a patient with Marfan's syndrome, thus compromising coronary blood flow producing ST segment depression. Continuous non-invasive aortic blood flow and PETCO2 monitoring proved valuable in the early detection and treatment of circulatory arrest and in the evaluation of the efficiency of peroperative CPR. PMID- 8706202 TI - Patient data management systems in anaesthesia: an emerging technology. AB - The purpose of this review is to define the expectations of an on-line automatic patient data management system (PDMS) into anaesthesia work-stations in and around the operating room suite. These expectations are based on review of available information in the medical literature, and trials of several systems that are available commercially, three of them in a more detailed fashion (i.e. Informatics, Datex and North American Drager). The ideal PDMS should: -- communicate with and capture the information from different monitors, anaesthesia machines and electronic gadgets (e.g., infusion pumps) used in the operating room (OR), while presenting selected relevant values and trends on a screen. -- inform the anaesthetist of deviations from preselected limits of physiological and technical values. In the future, the system will hopefully be upgraded to include an algorithm-based decision support system. -- communicate with the hospital mainframe computer, and automatically transfer demographic data, laboratory and imaging results, and records obtained during preoperative consultations. -- at the end of each anaesthetic procedure, create an anaesthetic record with relevant data automatically collected by the system, as well as that which was entered manually by the physician during the procedure. A copy of this anaesthesia file must be kept on a computerized archive system. None of the systems so far evaluated fulfilled all our expectations. We have therefore adopted approach for the gradual introduction of such a system into our OR environment over the next two to five years, during which expected improvements may be incorporated to upgrade the system. PMID- 8706199 TI - Side effects of intrathecal and epidural opioids. AB - The purpose of this article is to review the literature on the side effects of intrathecal and epidural opioids. English-language articles were identified through a MEDLINE search and through review of the bibliographies of identified articles. With the increasing utilization of intrathecal and epidural opioids in humans during the 1980s, a wide variety of clinically relevant side effects have been reported. The four classic side effects are pruritus, nausea and vomiting, urinary retention, and respiratory depression. Numerous other side effects have also been described. Most side effects are dose-dependent and may be more common if the opioid is administered intrathecally. Side effects are less common in patients chronically exposed to either intrathecal, epidural, or systemic opioids. Some side effects are mediated via interaction with specific opioid receptors while others are not. It is concluded that the introduction of intrathecal and epidural opioids marks one of the most important breakthroughs in pain management in the last two decades. However, a wide variety of clinically relevant non-nociceptive side effects may occur. All physicians utilizing intrathecal and epidural opioids must be aware of these side effects, for while most are minor, others are potentially lethal. PMID- 8706203 TI - Comparison of two administration techniques of inhaled nitric oxide on nitrogen dioxide production. AB - The purpose of this study was to verify whether, compared with the introduction of the NO-N2 mixture at the air inlet of the ventilator (classical method), a direct injection of NO-N2 into the inspiratory line of the ventilator circuit with a new injection device (new method), would reduce NO2 formation by reducing contact time between O2 and NO. The effect of two FIO2(0.21 and 0.90) and NO concentrations on NO2 production was determined. In the classical method, NO and O2 were mixed with an air/oxygen blender before the gas mixture entered the ventilator. In the new method, NO was injected directly into the respiratory line with the injection system. Nitric oxide and nitrogen dioxide gases were measured using a chemiluminescence analyzer. For a FI02 of 0.90 and 90 ppm of NO2, the amount of NO2 produced was decreased from 8.9 +/- 0.8 ppm (mean +/- SD) with the classical injection system to 4.4 +/- 0.2 ppm with the new injection system (P = 0.0039, Mann-Whitney test), and NO2 production was decreased from 4.5 +/-0.2 ppm to 2.1 +/- 0.4 ppm (P = 0.02) at 60 ppm of NO. However, at a FIO2, no difference was found in the amount of NO2 produced. We conclude that, compared with the classical method of NO administration, the new NO injection system reduces considerably the concentration of inhaled NO2 when a high FIO2 and a high concentration of NO are used. PMID- 8706204 TI - Choice of anaesthetic regimen influences haemodynamic response to cemented arthroplasty. AB - Haemodynamic changes during bilateral cemented arthroplasty (BCA) were compared in dogs anaesthetized with isoflurane/N2O (ISOF) or diazepam/fentanyl (100 microg x kg(-1))N2O(FENT). Eight animals were anaesthetized with each regimen. After establishing monitoring and recording baseline values, BCA was performed. Haemodynamic measurements included aortic blood pressure (ABP), pulmonary artery pressure (PAP), right and left atrial pressures, and cardiac output. These were recorded at 30, 60, 180 and 300 sec after BCA. Lungs were removed and examined postmortem using quantitative morphometry. Groups demonstrated similar increases in PAP (ISOF 15 +/- 2 to 32 +/- 7, FENT 19 +/- 4 to 38 +/- 13; P> 0.05 between groups, P< 0.05 vs baseline). The proportion of lung vasculature occluded by fat was not different between groups (ISOF 9.63 +/- 3.38%, FENT 8.85 +/- 2.20%). Stroke volume decreased similarly in both groups (P> 0,05 between groups, P< 0.05 vs baseline). However, ABP decreased within one minute of BCA in ISOF (111 +/- 17 to 55 +/- mmHg, P< 0.05 and two of eight dogs died. All FENT dogs survived and hypotension (118 +/- 20 to 102 +/- 24 mmHg) was transient and less severe (P< 0.05 vs ISOF). Increased heart rate (HR) was noted in FENT following BCA (73 +/- 8 to 108 +/- 25 beats x min(-1); P< 0.05). Baseline HR was higher in ISOF (P< 0.05) and no increase in HR was noted. Systemic vascular resistance decreased in ISOF (P< 0.05), but not FENT (P> 0.05 vs baseline, P< 0.05 vs ISOF). To assess the role of slower baseline HR in FENT (73 +/-8) versus ISOF (131 +/- 5), six FENT dogs were paced (130 beats x min(-1)) with epicardial leads and an AV sequential pulse generator to simulate the ISOF group's baseline HR. Haemodynamic stability was maintained in this group in spite of a more rapid baseline HR. The choice of anaesthetic regimen strongly influenced acute haemodynamic changes in response to BCA. PMID- 8706206 TI - Thiopentone inhibits beta-adrenergic responses in myocardial tissue. AB - The purpose of this study was to determine the importance of inhibition of beta adrenergic function in thiopentone-induced myocardial depression. Using an isolated, electrically stimulated rat left atria model, contractile dose-response curves to thiopentone (200 microM, 400 microM, 600 microM, 800 microM) were shifted to the right in preparations treated with 10(-3)M dibutyryl cyclic adenosine monophosphate (cAMP) compared with atria stimulated with 10(-6) M dibutyryl cyclic isoprenaline, demonstrating that inhibition of beta-adrenergic mechanisms by thiopentone is physiologically important. Depression by thiopentone was similar in atria treated with 10(-5) M forskolin compared with preparations stimulated with 10(-6) M isoprenaline, indicating that thiopentone does not block beta-adrenergic receptors. It is concluded that thiopentone depresses myocardial function by several mechanisms, one of which involves inhibition of the adenyl cyclase cascade. The adenyl cyclase enzyme is a likely site where thiopentone inhibits the system; however, other components of the cascade may also be involved. PMID- 8706207 TI - Fifty years in anaesthesia. PMID- 8706205 TI - Augmented sensitivity to benzodiazepine in septic shock rats. AB - The purpose of this study was to assess the pharmacological characteristics of the benzodiazepine binding site in the brain of septic animals. We induced endotoxin shock in rats using a caecum ligation and puncture model. Following examination of the physiological state of the rats 24 hr after the caecum ligation and puncture, brain tissue samples were prepared for biochemical assay of amino acids and for the [3H]-diazepam radioligand binding assay. Amino acids assays indicated that the concentration of aromatic amino acids was higher in the CLP group (P< 0.05), the branched chain amino acid concentration was lower in the CLP group (P< 0.05) and the sulfur-containing amino acid concentration was elevated in the CLP group (P< 0.05) than in both the control and the sham operated groups. [3H]-diazepam radioligand binding assays demonstrated that the number of receptors in the septic rats was increased in the forebrain (CLP rats; 2.37 +/- 0.04 pmol x mg(-1) protein, control rats; 1.45 +/- 0.02 pmol x mg(-1) protein, sham-operated rats; 1.49 +/- 0.03 pmol x mg(-1) protein), cerebellum (CLP rats; 1.55 +/- 0.05 pmol x mg(-1) protein, control rats; 1.05 +/- 0.02 pmol x mg(-1) protein, sham-operated rats: 1.09 +/- 0.02 pmol x mg(-1) protein) and brain stem (CLP rats; 1.21 +/- 0.04 pmol x mg(-1) protein, control rats; 0.61 +/- 0.02 pmol x mg(-1) protein, sham-operated rats; 0.63 +/- 0.02 pmol x mg(-1) protein) compared with the control and sham-operated rats (P< 0.05). In conclusion, it was considered that the increased number of benzodiazepine receptors may be one cause of the neuronal alteration observed in septic shock animals. PMID- 8706208 TI - Anaesthesia simulators. PMID- 8706209 TI - Anaesthesia simulators. PMID- 8706210 TI - Respiratory depression associated with patient-controlled analgesia. PMID- 8706211 TI - Filters and breathing circuit contamination. PMID- 8706212 TI - Management of autonomic dysfunction in severe tetanus: the use of fentanyl. PMID- 8706213 TI - Technology assessment: a Canadian perspective. PMID- 8706214 TI - Recovery room problems or problems in the PACU. PMID- 8706215 TI - Impaired haemostasis and regional anaesthesia. PMID- 8706216 TI - The diabetic patient: a challenge or just routine? PMID- 8706217 TI - Intravenous anaesthesia: new drugs, new concepts, and clinical applications. PMID- 8706218 TI - Pain management in children. PMID- 8706219 TI - The patient with a pacemaker or related device. AB - Patients with implanted pacemakers and ICDs can be safely managed for surgery and anaesthesia. Anaesthetic management of such patients should be planned first according to the patient's underlying medical status with particular emphasis on ventricular function and electrolyte balance. The anaesthetist must understand the various modes of pacemakers and ICDs available in the patient population. These devices are safe and well shielded form most electromagnetic interference in the operating room. Some precautions are nevertheless necessary. A magnet should not be placed routinely over a programmable pacemaker or ICD in the operating room, especially in the presence of electrocautery. Rate-responsive pacemakers should have rate adaptive modes disabled before surgery whenever possible. The mechanism of rate response should be known, so that inappropriate changes in heart rate can be avoided in the perioperative period if the rate responsive mode cannot for some reason be disabled. Antitachycardia pacemakers, should have the antitachycardia function disabled preoperatively. Methods for the provision of alternate emergency pacing should be available when dealing with patients at risk of bradyarrhythmias or pacemaker failure in the operating room. The anaesthetist should have a safe, practical plan of action that suites his/her experience and capabilities. ICDs should have automatic cardioverter defibrillator functions disabled for surgery and external modes of cardioversion/defibrillation should be available. PMID- 8706220 TI - Anaesthesia for major vascular surgery. PMID- 8706221 TI - New concepts in mechanical ventilation for ARDS. PMID- 8706222 TI - Monitoring tissue perfusion. PMID- 8706223 TI - Trends in neuroanaesthesia. PMID- 8706224 TI - Quality in anaesthesia: an integrated and constructive model. PMID- 8706225 TI - Anaesthesia and the transplanted patient. PMID- 8706226 TI - The patient with heart disease. PMID- 8706227 TI - HIV and AIDS: where is the epidemic going? AB - Routine surveillance of HIV (human immunodeficiency virus) infection and AIDS has been established over the past decade in many countries around the world. HIV estimates derived from empirical data are essential to the assessment of the HIV situation in different parts of the world and trends are used in tracking the development of regional epidemics, thereby keeping intervention activities focused on realities. As of the end of 1995, and following an extensive country by-country review of HIV/AIDS data, a cumulative total of 6 million AIDS cases were estimated to have occurred in adults and children worldwide and currently 20.1 million adults are estimated to be alive and infected with HIV or have AIDS. Of the total prevalent HIV infections, the majority remain concentrated in eastern, central and southern Africa, but the epidemic is evolving with spread of infection from urban to rural areas, as well as to West and South Africa, India and South-east Asia, and to a lesser extent--with proportional shifts to heterosexual infections--in North America, western Europe and Latin America. While the longer-term dimensions of the HIV epidemic at global level cannot be forecast with confidence, WHO currently projects a cumulative total of close to 40 million HIV infections in men, women and children by the year 2000. By that time, the male:female ratio of new infections will be close to 1:1. Recent trends indicate that HIV prevalence levels may be stabilizing or even decreasing among pregnant women in southern Zaire and parts of Uganda, among military recruits aged 21 in Thailand, and in some populations of northern Europe and the USA. While these changes may take place as part of the intrinsic dynamic of the epidemic, there is some evidence that declines in HIV prevalence are related to declines in HIV incidence which are, at least partly, due to prevention efforts. The challenge of surveillance and evaluation methods is now to identify the ingredients of success which may reveal a glimmer of hope. PMID- 8706228 TI - Treatment patterns for childhood diarrhoea: evidence from demographic and health surveys. AB - Treatment patterns for childhood diarrhoea among providers in public and private settings have been examined using data from 28 surveys in the Demographic and Health Surveys programme. In the majority of surveys, at least 50% of the children with diarrhoea who sought care from a health provider (public or private) received treatment that included oral rehydration salts (ORS). Private providers are a significant source of care for children with diarrhoea, but they are less likely to use ORS and more likely to prescribe unneeded drugs than providers in public settings. In countries where data are available, bloody diarrhoea appears to be undertreated. The results indicate that national public health programmes must continue to improve their strategies to ensure correct treatment of childhood diarrhoea by all health providers. PMID- 8706229 TI - Algorithms for verbal autopsies: a validation study in Kenyan children. AB - The verbal autopsy (VA) questionnaire is a widely used method for collecting information on cause-specific mortality where the medical certification of deaths in childhood is incomplete. This paper discusses review by physicians and expert algorithms as approaches to ascribing cause of deaths from the VA questionnaire and proposes an alternative, data-derived approach. In this validation study, the relatives of 295 children who had died in hospital were interviewed using a VA questionnaire. The children were assigned causes of death using data-derived algorithms obtained under logistic regression and using expert algorithms. For most causes of death, the data-derived algorithms and expert algorithms yielded similar levels of diagnostic accuracy. However, a data-derived algorithm for malaria gave a sensitivity of 71% (95% Cl: 58-84%), which was significantly higher than the sensitivity of 47% obtained under an expert algorithm. The need for exploring this and other ways in which the VA technique can be improved are discussed. The implications of less-than-perfect sensitivity and specificity are explored using numerical examples. Misclassification bias should be taken into consideration when planning and evaluating epidemiological studies. PMID- 8706230 TI - Prevalence of anti-Rift-Valley-fever IgM antibody in abattoir workers in the Nile delta during the 1993 outbreak in Egypt. AB - In the early summer of 1993, an outbreak of Rift Valley fever (RVF) was reported among both humans and animals in Aswan Governorate, Upper Egypt. To determine whether RVF infection had spread to the Nile delta region of the country, we carried out a cross-sectional survey of 1181 occupationally exposed abattoir workers (97% male; age 10-72 years) in 15 governorates of Egypt in November 1993. The overall prevalence of anti-RVF virus IgM antibody was 2% (range: 0% (7 governorates) to 10%). The highest prevalences were in Ismailia (10%) and Sharqiya (8%) Governorates. None of the seropositive subjects reported having experienced an episode of fever in the 2 months prior to the study. The prevalence of antibody was significantly higher (P < 0.05) among workers employed in high-risk jobs such as cutting animals' throats (relative risk (RR = 2.24)) and handling animal parts (RR = 2.37). The findings suggest that abattoir workers represent a useful sentinel population for surveillance of RVF. PMID- 8706231 TI - [Cercarial dermatitis in Europe: a new public health problem?]. AB - Cercarial dermatitis is a parasitic impasse that has worldwide distribution. The condition manifests itself as a highly pruriginous skin rash and is due to penetration of the dermis by larval stages (furcocercariae) of avian trematodes. Many species may be responsible for this disease. In Europe the genus Trichobilharzia is widely represented, in particular by the species T. ocellata; the definitive host is the duck (Anas platyrhinchos); the intermediate hosts are snails of the genus Lymnea (L. ovata or L. stagnalis). In France, cases of cercarial dermatitis were reported in June and July 1994 to the health authorities of three departements in the Pays de la Loire Region (western France). The epidemiological situation, common to the three maintained ponds that were concerned, is as follows: high level of eutrophication of the sites, colonization of the ponds by L. ovata, and settlement by many duck colonies. The simultaneous occurrence of these three phenomena, combined with long hours of sunshine in the summer, is responsible for most of the foci of cercarial dermatitis recently described in Europe. Control of this condition is difficult, requiring strict maintenance of bodies of water and if necessary the use of molluscicides such as niclosamide. The use of praziquantel in baits for treating the definitive hosts appears to interrupt the natural cycle of the avian Schistosomatidae. In the light of the observations reported here and the analysis of recent publications, cercarial dermatitis may be regarded as an emerging disease. Its public health impact needs to be evaluated at the global level. PMID- 8706232 TI - Perinatal mortality in rural Malawi. AB - Reported are the results of a study to assess the prevalence and risk factors for perinatal death among pregnant women in Malawi over the period 1987-90. There were 264 perinatal deaths among the 3866 women with singleton pregnancies (perinatal mortality rate, 68.3 per 1000 births). Among the risk factors for perinatal mortality were the following: reactive syphilis serology, nulliparity, a late fetal or neonatal death in the most recent previous birth, maternal height < 150 cm, home delivery, and low socioeconomic status. Although unexplained perinatal deaths will continue to occur, perinatal mortality can be reduced if its causes and risk factors in a community are given priority in antenatal and intrapartum care programmes. The following interventions could potentially reduce the perinatal mortality in the study population: screening and treating women with reactive syphilis serology; and management from early labour, by competent personnel in a health facility, of nulliparous women and multiparous women who are short or have a history of a perinatal death. PMID- 8706234 TI - A predictable comeback: the second pandemic of infections caused by Neisseria meningitidis serogroup A subgroup III in Africa, 1995. AB - Between 14 January and 4 April 1995 we isolated and characterized 44 meningococcal strains in Cameroon, Chad, Niger, and Burkina Faso; among these was the strain A:4:P1.9/clone III-1, which was involved in the second meningitis pandemic. This isolate was found in the clonal form in Niger and strains of the ET-37 complex were also found in the other three study countries, but apparently did not cause epidemics. One strain (Y:2a:P1.2,5 (ET-37 complex)) was isolated in January 1995 and another (A:4:P1.9) in March 1995 in Garoua (Cameroon). Eight strains were isolated in Moundou (Chad) between January and April 1995: the A:4:P1.9/clone III-1 (1 strain); members of the ET-37 complex (Y:2a:P1.2,5 (4 strains), Y:NT:P1.2,5 (1 strain), and Y:2a:-(1 strain)); and serogroup X (1 strain). In Niger, 31 strains were isolated between February and April 1995 from different regions. All were A:4:P1.9/clone III-1; between November 1994 and April 1995 there were 23814 cases of meningitis reported of which 2227 resulted in death. Three strains were isolated in Burkina Faso in April 1995: two were Y:2a:P1.2,5 (ET-37 complex) and one was A:4:P1.9/clone III-1. Thus in 1995 the epidemic and invasive strain (A:4:P1.9/clone III-1) responsible for the second pandemic was present in the four countries (Cameroon, Chad, Niger and Burkino Faso) that make up the area frequently affected by such epidemics and where cases are generally reported during the dry season. PMID- 8706233 TI - Breast-feeding counselling in a diarrhoeal disease hospital. AB - Lactation counsellors were trained to advise mothers of partially breast-fed infants who were admitted to hospital because of diarrhoea, so that they could start exclusive breast-feeding during their hospital stay. Infants (n = 250) up to 12 weeks of age were randomized to intervention and control groups. Mothers in the intervention group were individually advised by the counsellors while mothers in the control group received only routine group health education. During follow up at home by the counsellors a week later, only the mothers in the intervention group were counselled. All the mothers were evaluated for infant feeding practices at home two weeks after discharge. Among the 125 mother-infant pairs in each group, 60% of mothers in the intervention group were breast-feeding exclusively at discharge compared with only 6% in the control group (P < 0.001); two weeks later, these rates rose to 75% and 8% in the intervention and control groups, respectively (P < 0.001). However, 49% of mothers in the control group reverted back to bottle-feeding compared with 12% in the intervention group (P < 0.001). Thus, individual counselling had a positive impact on mothers to start exclusive breast-feeding during hospitalization and to continue the practice at home. Maternal and child health facilities should include lactation counselling as an integral part of their programme to improve infant feeding practices. PMID- 8706235 TI - Impact of oral rehydration and selected public health interventions on reduction of mortality from childhood diarrhoeal diseases in Mexico. AB - Reported are the results of an analysis of mortality trends from diarrhoeal diseases among under-5-year-olds in Mexico between 1978 and 1993 in relation to the impact of education, basic sanitation, and selected medical care practices. The study period was divided into three stages; the first pre-dated the widespread application of oral rehydration therapy (ORT); the second, covered the implementation of a nationwide programme promoting ORT; and the third included additional measures, such as immunization and improvements in basic sanitation. Mortality rates decreased progressively, at an average of 1.8% per year in the first stage, 6.4% in the second, and 17.8% in the third. The importance of literacy campaigns for women and the promotion of ORT was confirmed. Both of these measures reduced mortality; however, a greater reduction resulted from a massive immunization campaign against measles and improvements in sanitation (expansion of the drainage and piped water systems, improved water chlorination procedure, and effective prohibition of the use of sanitary sewage for vegetable irrigation). PMID- 8706237 TI - Data on birth weight in developing countries: can surveys help? AB - The main source of data on birth weight in developing countries is statistics from health facilities, although most developing countries do not produce annual estimates of the incidence of low birth weight from these data. Such estimates would be subject to selection bias as the data are usually limited to babies born within health facilities, and therefore are representative of a subgroup that is markedly different from the overall population of neonates. Since 1990 the Demographic and Health Surveys programme has included questions on recalled birth weight and relative size at birth in 15 national surveys. In this article, we show that these cross-sectional surveys can provide a useful data source for making national estimates of mean birth weight and the incidence of low birth weight. The extent of misclassification of birth weight is, however, too large to use the data on relative size at birth as an indicator of low birth weight at the individual level. PMID- 8706236 TI - Risk factors for childhood pneumonia among the urban poor in Fortaleza, Brazil: a case--control study. AB - Reported are the results of a case-control study carried out between July 1989 and June 1990 in Fortaleza city, Ceara State, Brazil, to determine the factors that place young children living in urban slum conditions at increased risk of contracting pneumonia. Cases were 650 under-2-year-olds with a radiological diagnosis of pneumonia who were recruited at the main paediatric hospital in the city over a full calendar year. Age-matched controls were recruited from the neighbourhood where the cases lived. Cases and controls were compared with respect to a variety of sociodemographic, environmental, reproductive, nutritional, and morbidity factors, and a risk factor questionnaire was administered to the mother of each child or to the child's normal guardian. Cases and controls were also weighed and measured. Malnutrition was the most important risk factor for childhood pneumonia in the study population, with weight-for-age, height-for-age, and weight-for-height also being important risk factors. In view of the high prevalence of stunting in the study population, there is an urgent need to reduce the level of malnutrition as a priority. Attendance at a day care centre was also associated with a high odds ratio. In view of the growing numbers of children attending day care centres in both developing and developed countries, it is essential that ways be identified to improve the design and management of such centres in order to minimize the risk of pneumonia. Increased risks of childhood pneumonia were also associated with low birth weight, non breast-feeding, crowding, high parity, and incomplete vaccination status, but not with socioeconomic status or environmental variables. Finally, children who had suffered from previous episodes of wheezing or been hospitalized for pneumonia had a greater than threefold increased risk of contracting the disease. PMID- 8706238 TI - Sonographic screening for urinary tract abnormalities in patients with Schistosoma haematobium infection: pitfalls in examining pregnant women. AB - In areas where Schistosoma haematobium is endemic, urinary schistosomiasis and pregnancy are frequently concomitant; however, both these conditions may produce similar urinary tract changes in ultrasound scans and hence their differential diagnosis may be difficult. In patients with urinary schistosomiasis, focal and/or diffuse urinary bladder wall changes are frequently detected ultrasonically. Dilatation of one or both ureters and progressive hydronephrosis may be observed in more severe cases. Satisfactory ultrasound examination of the urinary bladders of pregnant women is generally not feasible because mechanical compression by the fetus or transitory lower urinary tract infection hampers adequate filling of the bladder. Pregnancy itself is frequently associated with dilatation of one or both ureters and/or hydronephrosis; this is due to hormonal factors, infection, or compression of one or both ureters by the enlarged uterus and growing fetus. Hence, when sonography of the urinary bladder is not feasible such pregnancy-associated changes are virtually indistinguishable from those caused by S. haematobium, and may be incorrectly attributed to the latter. Pregnant women, therefore, should be excluded from ultrasonic surveys of urinary schistosomiasis. In contrast, ultrasound scans of adolescents and of women with positive parasitological findings and/or pathological alterations in the urinary tract should include examination of the uterus in order to assess whether the woman is pregnant; thereby, misinterpretation of sonographic findings can be avoided. Pregnant women with significant hydronephrosis must be closely followed up by an obstetrician since this condition may indicate a complication of the pregnancy; in some cases only a postpartum examination will permit definitive diagnosis. PMID- 8706239 TI - Why have mortality rates for severe malnutrition remained so high? AB - A review of the literature that has appeared over the past five decades indicates that the median case fatality from severe malnutrition has remained unchanged over this period and is typically 20-30%, with the highest levels (50-60%) being among those with oedematous malnutrition. A likely cause of this continuing high mortality is faulty case-management. A survey of treatment centres worldwide (n = 79) showed that for acutely ill children, inappropriate diets that are high in protein, energy and sodium and low in micronutrients are commonplace. Practices that could have fatal consequences, such as prescribing diuretics for oedema, were found to be widespread. Evidence of outmoded and conflicting teaching manuals also emerged. Since low mortality levels from malnutrition can be achieved using appropriate treatment regimens, updated treatment guidelines, which are practical and prescriptive rather than descriptive, need to be implemented as part of a comprehensive training programme. PMID- 8706241 TI - Reversibility of promoter induced hepatic focal lesion growth in mice. AB - The effect of cessation of phenobarbital and dieldren treatment on hepatic focal lesion growth in male B6C3F1 mice was investigated. Following induction of lesions by diethylnitrosamine, mice were placed on control NIH-07 diet (control diet) or NIH-07 diet containing either dieldrin (10.0 mg/kg diet) or phenobarbital (500 mg/kg diet). Mice were sacrificed after 30 and 60 days of dietary treatment. Two additional groups of mice were fed either the dieldren- or phenobarbital-containing diet for 30 days followed by feeding of NIH-07-only diet for an additional 30 days. The effect of treatment and removal of dieldrin or phenobarbital on lesion growth was examined by measuring both the number of focal lesions per liver and the relative volume of focal lesions. In addition, the rate of cell proliferation and programmed cell death in focal lesion growth was investigated by examining DNA synthesis and apoptosis in the focal lesions. Dietary dieldrin or phenobarbital increased the number of focal lesions and the focal lesion volume. In both dieldrin- and phenobarbital-treated mice, an increased number of eosinophilic lesions were seen. The focal lesion volume was increased in both eosinophilic and basophilic lesions. Dieldrin and phenobarbital treatment also increased the DNA synthetic labeling index in both eosinophilic and basophilic lesions. Removal of dieldrin or phenobarbital from the diet after 30 days of promoter treatment decreased the total number and volume of hepatic focal lesions. The labeling index of the focal lesions was also decreased in these mice. At the terminal sacrifice, the percentage of apoptotic cells in focal lesions was higher in mice fed dieldrin- or phenobarbital-containing diets for the entire 60 days than in mice returned to control diet for the last 30 days. Eosinophilic lesions were more dependent on the presence of a promoting stimulus than the basophilic lesions. These data indicate that induction and maintenance of the growth of some preneoplastic lesions in the mouse may be dependent upon continuous tumor promoter treatment. PMID- 8706240 TI - The human papillomavirus E6/E7 genes induce discordant changes in the expression of cell growth regulatory proteins. AB - The E6/E7 oncoproteins of human papillomavirus (HPV) types 16 and 18 are responsible for the efficient immortalization of human genital keratinocytes and we have recently reported that such immortalized cells display alterations in the expression of cyclin A, cyclin B, and cdc-2. To determine whether these alterations were the consequence of E6/E7 protein expression or whether they resulted from the process of cellular immortalization, we multiply-infected primary genital keratinocytes with a retrovirus expressing the HPV-18 E6/E7 genes and examined the cells for acute, pre-immortalization changes in several critical cell growth regulatory proteins including cyclin A, cyclin B, cdc-2, p53 and c myc. In addition, we simultaneously evaluated the expression of the E6/E7, bcl-2 and involucrin genes to determine whether there were accompanying alterations in the expression of viral genes or in cellular genes related to cell apoptosis and the state of keratinocyte differentiation. The cell cycle regulating proteins (cyclin A, cyclin B, cdc-2 and p53) change significantly within days after retroviral infection. Cyclin B and cdc-2 increase over 4-fold by three passages and remain relatively constant thereafter through passage 21, whereas the levels of p53 protein decrease 25% by passage three. Increases in the expression of cyclin A, cyclin B and cdc-2, and decreases in p53 are therefore among the earliest observable changes in cell regulatory proteins following E6/E7 gene expression and may be important contributors to the development of cell immortalization. The expressions of viral E6/E7 genes, c-myc, bcl-2 and involucrin exhibit progressive changes with increased passage numbers until passage 21, presumably reflecting the selective outgrowth of immortalized cells. PMID- 8706242 TI - Effects of dietary beta-carotene and selenium on initiation and promotion of pancreatic carcinogenesis in azaserine-treated rats. AB - In the present study the effects of 0.1 or 1.0 g beta-carotene/kg diet (L beta C or H beta C) and 1.0 mg or 2.5 mg selenium/kg diet (LSel or HSel), as well as combinations of the respective low and high concentrations of beta-carotene and selenium (LMix or HMix) on the initiation/early promotion phase or on the late promotion phase of pancreatic carcinogenesis in azaserine-treated rats, were investigated using cell proliferation and volumetric data of atypical acinar cell foci (AACF) as parameters. The present results indicate chemopreventive effects of dietary selenium, dietary beta-carotene and of their combination on the development of acinar pancreatic lesions induced in rats by azaserine. The inhibitory effect was most pronounced when beta-carotene and/or selenium were added to the diets during the late promotion phase of the carcinogenic process, although inhibition was also observed with these compounds when they were added to the diets during the first 5 weeks of the study only (initiation/early promotion phase). Neither in the initiation/early promotion phase nor in the late promotion phase was a dose-related trend observed. The multiplicities of AACF with a diameter over 1.0 mm and of carcinomas in situ (CIS), as well as the incidence of CIS were not significantly different among the groups. However, in the late promotion experiment a dose-related decline in multiplicity could be observed in the selenium supplemented groups and in the groups receiving combinations of beta-carotene and selenium. Cell proliferation in azaserine induced AACF, as estimated by the bromodeoxyuridine (BrdU) labeling index, was significantly higher in H beta C, HSel, LMix and HMix groups (initiation/early promotion phase) as well as in H beta C, LSel, HSel, LMix and HMix groups (late promotion phase) than in high fat controls. From the present results it can be concluded that: (i) beta-carotene and selenium have inhibitory effects on pancreatic carcinogenesis induced in rats by azaserine; (ii) the most clear effects were observed when selenium was given as such, or in combination with beta-carotene during the late promotion phase; and (iii) beta-carotene and selenium stimulate cell proliferation in AACF. PMID- 8706243 TI - Drug-resistant breast cancer cells frequently retain expression of a functional wild-type p53 protein. AB - Abnormalities in the p53 tumor suppressor gene have been shown to affect cellular processes related to cell cycle control and gene amplification. In this study we compare the status and function of wild-type p53 in MCF-7 breast cancer cells with sublines selected for resistance to chemotherapeutic agents having different mechanisms of action. Sublines that were resistant to melphalan, pyrazafurin, mitoxantrone, etoposide and PALA all retained expression of wild-type p53. Methotrexate-resistant MCF-7 cells were unusual heterozygotes that expressed a wild-type and dominant, in-frame p53 deletion mutant and the doxorubicin resistant cells expressed only mutant p53. Analysis of the G1 checkpoint after treatment with ionizing radiation revealed that the pyrazafurin-, melphalan- and mitoxantrone-resistant cells arrested strongly in G1. The etoposide- and PALA resistant cells had an intermediate G1 arrest phenotype and the methotrexate- and doxorubicin-resistant cells had a minimal G1 arrest phenotype. mRNA and protein analyses of downstream effector genes, including P21CIP1/Waf1, mdm2, Gadd 45 and the retinoblastoma protein, did not entirely differentiate sublines having a strong versus intermediate G1 arrest phenotype. Neither the p53 status nor the strength of the G1 arrest could be correlated with cell survival after ionizing radiation. When drug-sensitive MCF-7 cells were treated with the same chemotherapeutic agents, p53 and p21CIP1/Waf1 levels increased between 2- and 14 fold. Together these data suggest that other cellular factors likely play a role in overcoming the inhibitory effects of ionizing radiation on p53 in drug resistant breast cancer cells. PMID- 8706244 TI - Protection by green tea, black tea, and indole-3-carbinol against 2-amino-3 methylimidazo[4,5-f]quinoline-induced DNA adducts and colonic aberrant crypts in the F344 rat. AB - Male F344 rats were exposed for 8 weeks to extracts of green tea (2% w/v) or black tea (1% w/v), or to 0.1% dietary indole-3-carbinol (I3C). In weeks 3 and 4 of the study, rats were given 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) every other day by oral gavage (50 mg/kg body wt) in order to induce aberrant crypt foci (ACF) in the colon. Compared with controls given IQ alone, all three inhibitors reduced the number of total aberrant crypts per colon, and green tea and I3C inhibited significantly the mean number of ACF (P < 0.05). Rats pre treated with green tea, black tea, or I3C and given a single p.o. injection of 50 mg IQ/kg body wt 24-48 h before sacrifice had reduced levels of IQ-DNA adducts in the liver, and excreted lower amounts of IQ and other promutagens in the urine and feces. Inhibitors also reduced the excretion of IQ-sulfamate in the urine, but increased the relative amounts of IQ-5-O-sulfate and IQ-5-O-glucuronide. Western blotting together with assays for 7-ethoxyresorufin O-deethylase and methoxyresorufin O-demethylase established that I3C preferentially induced cytochrome P4501A1 over 1A2, consistent with the altered profile of urinary metabolites. However, both teas caused slight induction of cytochrome P4501A2 versus 1A1, which would be predicted to enhance the activation of IQ. Thus, green tea and black tea are likely to protect against IQ-DNA adducts and ACF by mechanisms other than induction of cytochromes P450, such as inhibition of enzymes which activate IQ or the scavenging of reactive intermediates. PMID- 8706245 TI - Differential activity of aspirin, ketoprofen and sulindac as cancer chemopreventive agents in the mouse urinary bladder. AB - In vivo studies were conducted to compare the activity of three non-steroidal anti-inflammatory drugs as inhibitors of urinary bladder carcinogenesis induced in B6D2F1 (BDF) mice by N-butyl-N-(4-hydroxybutyl)nitrosamine (OH-BBN). Mice received continuous dietary exposure to non-toxic doses of aspirin, sulindac or ketoprofen beginning 1 week prior to the first of eight weekly doses of 7.5 mg OH BBN; studies were terminated at 24 weeks after the first carcinogen dose. Both dose levels of sulindac (200 and 400 mg/kg diet) and both dose levels of ketoprofen (40 and 80 mg/kg diet) reduced the incidence of transitional cell carcinoma of the urinary bladder by >70% from that seen in dietary controls. The high dose of sulindac conferred the greatest protection against bladder cancer induction. In contrast, when administered at 400 and 800 mg/kg diet aspirin was inactive as a chemopreventive agent in the OH-BBN/BDF bladder cancer model. The significant potency of sulindac and ketoprofen as inhibitors of urinary bladder carcinogenesis, when considered with their history of safe human use, suggests that these agents merit further study as drugs for cancer chemoprevention in this target tissue. PMID- 8706246 TI - Effect of oltipraz, alpha-tocopherol, beta-carotene and phenethylisothiocyanate on rat oesophageal, gastric, colonic and hepatic glutathione, glutathione S transferase and peroxidase. AB - Four anticarcinogens (oltipraz, alpha-tocopherol, beta-carotene and phenethylisothiocyanate [PEITC]) were studied with respect to their effects on oesophageal, gastric, colonic and hepatic (i) glutathione (GSH) content, (ii) glutathione S-transferase (GST) enzyme activity, (iii) GST isoenzyme levels, and (iv) glutathione peroxidase (GPx) enzyme activity in male Wistar rats. GST enzyme activity was significantly increased in oesophagus (1.9X) and colon (1.2X) by PEITC and in liver (1.4X) by oltipraz. GST Alpha was doubled in the liver by oltipraz, alpha-tocopherol and PEITC. GST Mu levels were increased by beta carotene and PEITC in stomach and liver, by oltipraz in liver and by alpha tocopherol in stomach. PEITC induced colonic GST Pi levels (1.3X). GSH content was induced in liver by oltipraz (1.4X) and alpha-tocopherol (1.2X) and in colon by PEITC (1.6X). Each of the anticarcinogens tested increased GPx activity at one or more sites: Se-dependent and total GPx activities were induced in 31.3% and 37.5% of all possibilities, respectively. Major induction in total GPx was found in stomach by alpha-tocopherol (1.8X). In conclusion our data demonstrate that dietary administration of oltipraz, PEITC, alpha-tocopherol and beta-carotene, may exert chemopreventive effects in the digestive tract of the rat by enhancing GST, GPx, and, to a lesser extent, GSH levels. PMID- 8706247 TI - Transferrin C3 offers protection against smoking-associated lung cancer? AB - In previous investigations increased body iron stores and transferrin (TF) variants have been found to be associated with adverse health effects, including cancer. In this investigation transferrin C (TF C) subtypes were studied in lung cancer patients and controls from the Stockholm area in central Sweden. There was a significant difference between patients and controls with respect to the distribution of TF C alleles and genotypes, which was mainly due to a low frequency of the TF C3 allele among the patients (P = 3 x 10(-6). However, in adenocarcinoma the frequency of TF C3 types was almost identical to that among the controls, whereas in the smoking-related (squamous and small cell) tumor types the TF C3 frequency was remarkably low (OR = 0.03, 95% CI = 0.00-0.22). Thus individuals with the TF C3 variant appear to enjoy an almost complete protection against smoking-related lung cancer. The frequency of individuals carrying the protective TF C3 variant is approximately 17% in central Sweden and 25% in Finland, which has the highest TF C3 frequency found so far. The mechanism behind the observed association, which appears to be independent of iron binding and body iron stores, remains to be elucidated. PMID- 8706248 TI - Prepubertal genistein exposure suppresses mammary cancer and enhances gland differentiation in rats. AB - Genistein, a component of soy, was administered to prepubertal female Sprague Dawley CD rats and investigated for chemoprevention against mammary cancer. Genistein, at 500 microgram/g body wt or an equivalent volume of the vehicle, dimethylsulfoxide (DMSO), was injected (s.c.) on days 16, 18 and 20 post-partum. At day 50 post-partum all animals were exposed to 80 microgram dimethylbenz[a]anthracene (DMBA) per g body wt. Animals treated prepubertally with genistein as compared to DMSO had reduced incidence and significantly fewer adenocarcinomas per animal. Mammary whole mount analysis showed that prepubertal genistein treatment resulted in mammary glands of 50-day-old rats developing fewer terminal end buds and more lobules II. Cell proliferation studies with bromodeoxyuridine (BrdU) showed that terminal end buds from mammary glands of 50 day-old females treated prepubertally with genistein had significantly fewer cells in S-phase of the cell cycle. Serum genistein concentrations in 21- and 50 day-old females following prepubertal genistein treatment were 4.2 +/- 0.6 micromol and 102 +/- 30 nM, respectively. Animals treated prepubertally with genistein as compared to vehicle spent more time in the estrus phase of the estrus cycle, although all animals did cycle. In 50-day-old females, circulating estradiol-17 beta and progesterone concentrations were not significantly altered by the prepubertal genistein treatment. Oocyte/follicle counts and numbers of atretic follicles and corpora lutea were not significantly different between the genistein- and vehicle-treated animals. We conclude that genistein treatment during the prepubertal period can suppress the development of chemically-induced mammary cancer without significant toxicity to the endocrine/reproductive system. PMID- 8706249 TI - Modulatory influence of chlorophyllin on the mouse skin papillomagenesis and xenobiotic detoxication system. AB - The present study evaluates the modulatory potential of chlorophyllin (CHL) on the murine skin papillomagenesis pattern and its influence on the levels of biotransformation system enzymes. Topical application of CHL (100 mg/kg body weight/day) during peri-, post- or peri- and post-initiational stages of 7,12 dimethylbenz[a]anthracene (DMBA)-induced papillomagenesis, significantly (P < 0.01) reduced the (i) tumor burden to 3.68, 3.56 and 3.33 (positive control value: 5.89); (ii) cumulative number of papillomas to 59, 57 and 60 (positive control value: 112); and (iii) incidence of mice bearing papillomas to 88%, 88% and 90%, respectively (positive control value 100%). CHL treatment alone or during peri-, post-, or peri- and post-initiational stages significantly elevated the glutathione S-transferase (GST) and -SH levels in the liver and skin tissue of the murine system. The potential of CHL in modulating the process of carcinogenesis is suggested by the altered levels of biotransformation system enzymes. The implications of the biochemical changes and inhibition of tumor incidence by CHL are discussed. PMID- 8706250 TI - DNA adduct formation by allyl glycidyl ether. AB - Glycidyl ethers are reactive epoxides used as components of a variety of epoxy materials. These compounds are known to cause allergic reactions, but since they are generally also genotoxic it would be of interest to evaluate the risk for induction of such effects. Reaction products of allyl glycidyl ether with nucleic acid components were therefore studied. Adduct standards of expected major products in DNA were prepared and assigned to N-7-guanine, N-1- and N-3-adenine and N-3-cytosine. The adducts were characterized by UV spectroscopy, and the adduct to N-1-adenine also by mass spectrometry and nuclear magnetic resonance spectroscopy. In analogy with the formation of corresponding reaction products of other simple epoxides the N-1-adenine adduct rearranged in a base catalysed reaction to N6 and the N-3-cytosine adduct deaminated to form the corresponding N 3-uracil adduct. For allyl glycidyl ether these further reactions of the N-1 adenine and N-3-cytosine adducts were, however, slower than has been observed for corresponding products of other epoxides, but faster than for methylated and ethylated products. In double-stranded salmon testis DNA treated in vitro with allyl glycidyl ether, the major product was found at N-7-guanine, followed by those at N-1-adenine, N-3-adenine and N-3-cytosine (including N-3-uracil). A minor amount of an N6-adenine adduct was also detected, but only after 48 h of reaction. In single-stranded DNA the yield of the N-1-adenine adduct was increased to about the level of the N-7-guanine adduct. The level of the N-3 cytosine adduct was also considerably higher in single-stranded DNA and was the third largest adduct. The reactivity of N-3-adenine was decreased in single stranded DNA and since other adducts increased the relative yield of this adduct was very low. The N-7-guanine and N-3-adenine adducts were lost from DNA as a consequence of depurination with half-lives in double-stranded DNA at 37 degrees C and pH 7.4 of 38 and 20 h, respectively. The rates of losses (due to depurination or rearrangement) of initially formed adducts in DNA increased in the order N-1-adenine < N-7-guanine approximately N-3-cytosine < N-3-adenine and were faster in single- than in double-stranded DNA. Taking only the rate of formation and chemical stability into consideration, the adducts with N-1-adenine and N-7-guanine seem to be the most promising candidates for monitoring allyl glycidyl ether exposures in vivo. PMID- 8706251 TI - Carcinogenicity of cyproterone acetate in the mouse. AB - The synthetic progestin cyproterone acetate (CPA) has been shown to be a hepatocarcinogen in the rat, but little is known of its effects in mice. A 52 week CPA study in the mouse strain C57Bl/10J has been reported not to produce liver tumours, although CPA induced significant liver enlargement and induction of the mixed function oxidase CYP3A. The present study is a further investigation of the effects of CPA in mice of the C57Bl/10J strain dosed for 104 weeks. A group of 40 mice/sex were fed 800 p.p.m. CPA in the diet for 104 weeks with a control group of eight/sex. Mortality was high in females after 40 weeks due to hormonal effects in the uterus; no female and only four CPA-dosed males survived to 104 weeks. Liver cell hypertrophy with increased fat and glycogen and single cell or small multifocal areas of hepatocellular necrosis were universal. Proliferating cell nuclear antigen demonstrated an increase in proliferating cells within tumours and within the non-tumour bearing liver of CPA-dosed mice compared with normal livers of control mice. Hepatocellular tumours developed in 44% of males and 22% of females dosed with CPA, compared with none in the controls (the strain has a low, <10%, incidence of spontaneous liver tumours compared with other mouse strains). In addition, over 85% of both sexes dosed with CPA developed adenomatous polyps of the pyloric antrum and pancreatic islet cell hyperplasia, shown by immunostaining to be chiefly of insulin-secreting cells. Adrenocortical atrophy was also observed with other widespread effects in the endocrine system. The results suggest that the liver tumours, as in the rat, are likely to be related to effects on liver growth and mitogenesis. It is suggested that the tumours of the stomach and the pancreatic islet cell hyperplasia are manifestations of the effects of CPA in the endocrine system. PMID- 8706252 TI - Expression of cytochrome P450s and glutathione S-transferases in human esophagus with squamous-cell carcinomas. AB - In order to clarify the expression of cytochrome P450 and glutathione S transferase in human esophagus, 41 samples of human esophagus with squamous-cell carcinoma were investigated by immunoblot analysis and enzyme assays. Cytochrome P450 1A2/1 was clearly expressed in microsomes, and the amount in samples with tumorous tissue was significantly greater than that in samples without tumourous tissues or in liver; cytochrome P450 2B6 and 3A4/3 were expressed polymorphically. Aryl hydrocarbon hydroxylase activity was detected in microsomes and was greater in samples from smokers than non-smokers. Patients who both smoked and drank alcohol, however, had activity similar to that of patients without these habits. Glutathione S-transferase M1 and A1/2 protein existed polymorphically in cytosol, and glutathione S-transferase P1-1 was detected in all samples. The frequency of expression of the glutathione S-transferase A1/2 protein was greater in patients with M1 protein than in those without; no difference in the expression was seen for glutathione S-transferase P1-1. Neither smoking nor drinking influenced the expression or activity of glutathione S transferase. Our data support the idea that some carcinogens can be directly activated or inactivated in human esophageal epithelium. PMID- 8706253 TI - In vivo formation of mutagens by intraperitoneal administration of polycyclic aromatic hydrocarbons in animals during exposure to nitrogen dioxide. AB - Consumption of fossil fuels has increased indoor and outdoor concentrations of polycyclic aromatic hydrocarbons (PAHs) and nitrogen dioxide (NO2). To study the combined effect of PAH administration and NO2 exposure on mutagenicity of urine from animals we injected 400 mg/kg body wt i.p. one of five kinds of PAH (pyrene, fluoranthene, fluorene, anthracene and chrysene) into ICR mice, Wistar rats, Syrian golden hamsters or Hartley guinea pigs after exposure to 20 p.p.m. NO2 gas for 24 h and then exposed the animals to NO2 gas for an additional 24 h. During the latter 24 h we collected the urine and assayed its mutagenicity with the Ames Salmonella strains after treatment with beta-glucuronidase and arylsulfatase and extraction with dichloromethane. The urine from mice treated with both PAH and NO2 showed high mutagenicity for Salmonella typhimurium strains TA98 and TA100, whereas the urine from mice treated with PAH and air showed almost no mutagenic activity. The mutagenicity was decreased in nitroreductase- and acetyltransferase deficient strains TA98NR and TA98/1,8-DNP6 respectively. Treatment with a mixture of 20% of each of the five kinds of PAH and NO2 augmented the urinary mutagenicity of mice 1.5-fold. The urine from hamsters treated with pyrene or fluoranthene and NO2 was also highly mutagenic, but that from rats or guinea pigs was not very mutagenic. The mutagenicity was also decreased in strains TA98NR and TA98/1,8-DNP6. These results suggest that the urine contains nitro compounds and that the nitration of PAHs occurs in the body of animals under exposure to NO2 gas. Actually, the nitrated metabolites of pyrene, 1-nitro-6/8-hydroxypyrene and 1-nitro-3-hydroxypyrene, were detected in the urine from mice treated with pyrene under exposure to NO2 gas. To elucidate the mechanism of in vivo nitration, NO2 (20 p.p.m.) was bubbled through 50 mM Tris-HCl buffer (pH 7.4) or dichloromethane solution containing pyrene or 1-hydroxypyrene (10 microg/ml). Pyrene was not nitrated by NO2 in either aqueous or organic solutions. However, 1-hydroxypyrene was changed to nitrohydroxypyrenes by NO2 in the Tris-HCl buffer, but not in the organic solution. Ascorbic acid, alpha-tocopherol, glutathione oleic acid and hemoglobin were found to inhibit the nitration of 1-hydroxypyrene in aqueous solution. The urinary mutagenicity of mice treated with both pyrene and NO2 was also decreased by oral administration of ascorbic acid and alpha-tocopherol. These results suggest that 1-hydroxypyrene is nitrated by an ionic reaction in the animal body after hydroxylation of pyrene in the liver. PMID- 8706254 TI - Glutathione S-transferase A1-1-catalysed conjugation of bay and fjord region diol epoxides or polycyclic aromatic hydrocarbons with glutathione. AB - Mammalian metabolism of polycyclic aromatic hydrocarbons results in the formation of vicinal diol epoxides (existing as enantiomeric pairs of two diastereomers) considered as important ultimate carcinogens if the oxirane ring is located in a bay or fjord region of the parent hydrocarbon. In the present study, individual stereoisomers of the bay region diol epoxides of chrysene, dibenz[a,h]-anthracene and benzo[a]pyrene, as well as of the fjord region diol epoxides of benzo[c]phenanthrene, benzo[c]chrysene and benzo[g]chrysene, have been incubated with glutathione (GSH) in the presence or absence of human glutathione S transferase isoenzyme GST A1-1, a class Alpha enzyme. The formation of GSH conjugates was determined and quantified by HPLC. The results demonstrate that the GST A1-1 isoenzyme catalyzes the formation of GSH conjugates of all diol epoxides tested, although a marked variation in catalytic efficiency (>20-fold) was observed. With both bay and fjord region anti-diol epoxides a significant preference for conjugation of the enantiomer with the R configuration at the benzylic position of the oxirane ring was noted. Among the syn diastereomers of the fjord region diol epoxides a similar substrate enantioselectivity was noted, i.e. the enantiomer with the corresponding R configuration was again preferentially conjugated. In contrast, for the bay region syn-diol epoxides this substrate selectivity was reversed, resulting in a preference for the enantiomer with the S configuration. The chemically more reactive syn diastereomers were in general better substrates for GST A1-1 than the corresponding anti diastereomers. However, a comparison between different diol epoxide diastereomers revealed no obvious correlation between chemical reactivity of the compounds and catalytic efficiencies. Furthermore, no significant correlation between diol epoxide lipophilicity and catalytic efficiency was observed. It is suggested that stereochemical factors, including the size and the geometry of the aromatic ring system and the preferred conformation of the diol epoxide, are involved as the major determinant for the rate of catalysis by GST A1-1. PMID- 8706255 TI - Formation of DNA adducts by the co-mutagen norharman with aromatic amines. AB - Norharman, widely distributed in our environment, is alone not mutagenic to Salmonella typhimurium TA98 and TA100 either with or without S9 mix, but becomes mutagenic to S.typhimurium TA98 with S9 mix when non-mutagenic aromatic amines like aniline or o- or m-toluidine are added. Thus norharman has been called a 'co mutagen'. In the present study we examined whether or not DNA adducts are formed in DNA of S.typhimurium TA98 by treatment with norharman and aromatic amines using 32P-post-labeling analysis under modified adduct intensification conditions. When a sample of norharman (8 mg) and aniline (4 mg) was incubated with 4 ml of overnight culture of S.typhimurium TA98 in the presence of 20 ml S9 mix for 6 h at 37 degrees C, three adduct spots were detected at a total relative adduct labeling (RAL) of 10.8 +/- 2.27/10(8) nucleotides. Under the same conditions, a mixture of norharman (8 mg) and o-toluidine (4 mg) yielded three adduct spots at a RAL of 3.74 +/- 1.71/10(8) nucleotides. With a combination of norharman and m-toluidine, a single adduct spot was seen at a RAL of 0.04 +/- 0.01/10(8) nucleotides. In contrast, norharman with p-toluidine did not produce adduct spots. Furthermore, neither norharman nor the aromatic amines themselves gave any evidence of adducts. Thus DNA adduct formation by norharman with aromatic amines correlates with the co-mutagenic action of norharman in S.typhimurium TA98. PMID- 8706256 TI - Bilioenterostomy enhances biliary carcinogenesis in hamsters. AB - The aim of this study was to examine whether the type of bilioenterostomy enhances biliary carcinogenesis in the hamster model. Syrian hamsters were divided into the following groups; simple laparotomy (control group), cholecystoduodenostomy with dissection of the extrahepatic bile duct on the distal end of the common duct (CDDB group) and cholecystoileostomy with dissection of the extrahepatic bile duct on the distal end of the common duct (CIDB group). Following these procedures, all hamsters received N-nitrosobis(2 oxopropyl)amine. The diameter of the extrahepatic bile duct and plasma levels of cholecystokinin (CCK) were measured and the number of neoplastic lesions was counted microscopically. Proliferative effect of the procedures on the biliary epithelium was examined by proliferative cell nuclear antigen. In the CDDB group the extrahepatic bile duct was significantly dilated and carcinogenesis of the gall-bladder and extrahepatic bile ducts was enhanced. In the CIDB group the CCK bioactivity was stimulated and intrahepatic biliary duct, but not gall bladder and extrahepatic bile duct, carcinogenesis was promoted more than that observed in the CDDB group. Proliferation of the biliary duct epithelium was enhanced in both the CDDB and CIDB groups. Cholecystoduodenostomy enhanced intra- and extrahepatic bile duct carcinoma, whereas cholecystoileostomy promoted only intrahepatic bile duct carcinoma. Some factors in the intestinal juice seem to play a role in the promotion of biliary tract carcinoma. PMID- 8706257 TI - Chromium(VI) treatment of normal human lung cells results in guanine-specific DNA polymerase arrest, DNA-DNA cross-links and S-phase blockade of cell cycle. AB - Previous studies have shown that in vitro treatment of a synthetic double stranded DNA template with chromium(III), or chromium(VI) in the presence of ascorbate, resulted in guanine-specific DNA polymerase arrests that correlated strongly with DNA-DNA cross-linking. In vivo chromium(VI) undergoes a more complicated intracellular cascade of reductive metabolism than is achievable in an in vitro model. Moreover, in living cells, DNA is highly packaged in the form of chromatin which may alter the accessibility of DNA to chromium. A repetitive primer-extension assay was employed to determine whether chromium forms polymerase-arresting lesions in vivo. Normal human lung fibroblasts treated with chromium(VI) exhibited adduct levels of 0.13-0.92 mmol Cr/mol DNA-nucleotides in the total genome (0.26-1.84 Cr adducts/Kbp DNA) and DNA interstrand cross-links. Genomic DNA was isolated and alphoid sequences (1-5% of the genome) were used as a substrate for repetitive primer extension using Taq polymerase. The results showed a dose-dependent, guanine-specific, replication termination, even at low doses resulting in greater than 90% survival. The same treatment resulted in dose dependent suppression of thymidine incorporation into DNA immediately after treatment. Thymidine incorporation increased during the first 6 h after the 2-h exposure, probably related to the repair of the single strand breaks, but then returned to high suppression levels at 24 h. The chromate treatments inhibited cell growth by specific blocking of the progression of cells through S-phase of the cell cycle. The results confirmed our studies in cell-free systems and taken together they strongly indicate that guanine-guanine DNA interstrand cross-links induced by chromate in living cells is the lesion responsible for blocking DNA replication processivity. PMID- 8706258 TI - Mouse lung tumors exhibit specific Ki-ras mutations following transplacental exposure to 3-methylcholanthrene. AB - A pharmacogenetic mouse model was utilized to determine the role of Cyp1a1 expression on the formation of Ki-ras mutations in lung tumors following transplacental exposure to polycyclic aromatic hydrocarbons (PAHs). A backcross between Ah responsive male B6D2F1 mice and nonresponsive female DBA mice resulted in a litter in which both responsive and nonresponsive fetuses resided in the same nonresponsive maternal environment. Pregnant mothers received a single i.p. injection of either 10 or 30 mg/kg of 3-methylcholanthrene (MC) or olive oil vehicle on day 17 of gestation. At the higher dose of MC, the responsive offspring of both sexes had significantly (P < 0.05) higher incidences of lung tumors than their nonresponsive littermates. The male responsive mice also exhibited a significantly increased liver tumor incidence over the nonresponsive mice at the P < 0.05 level. Administration of 10 mg/kg of MC caused a very low incidence of lung tumors and did not result in the appearance of macroscopically visible liver tumors. Exons 1 and 2 of the Ki-ras gene were amplified from paraffin-embedded tissue samples. The PCR products were screened by allele specific oligonucleotide hybridization (ASO). Thirteen of 16 lung tumors (81%) screened exhibited point mutations in the 12th or 13th codon, including seven tumors that contained GGT-->GTT (GLY12-->VAL12) transversions, four which exhibited GGT-->TGT (GLY12-->CYS12) transversions, and two which contained GGC- >CGC (GLY13-->ARG13) transversions. None of the tumors had mutations at codon 61. The results obtained by ASO were confirmed by cloning and sequence analysis of the PCR products from four of these tumors. Within the subset of 16 tumors examined in this study, the same types of mutations in the Ki-ras gene were generally present in both responsive and nonresponsive mice, although G-->C transversions were found in two tumors from a single responsive female mouse. Interestingly, while both males and females exhibited the GGT-->GTT mutations at codon 12, the GGT-->TGT transversion was only found in male mice. These results are consistent with a key role for Cyp1a1 in modulating individual susceptibility to cancer formation through the formation of reactive intermediates that bind to DNA and result in activating mutations in key regulatory molecules. PMID- 8706259 TI - Inhibition of gap junctional intercellular communication and delocalization of the cell adhesion molecule E-cadherin by tumor promoters. AB - The effect of 12-O-tetradecanoylphorbol-13-acetate (TPA) and benzoyl peroxide (BoP) on gap junctional intercellular communication (GJIC) and the amount and localization of E-cadherin was studied in initiated mouse epidermal cells (3PC) and in carcinoma cells (CA3/7) originating from the same cell type. In addition, the localization and phosphorylation of connexin43 was studied in both cell lines and in primary keratinocytes. GJIC inhibition by TPA and BoP was stronger in primary keratinocytes compared with both cell lines. BoP strongly decreased the amount of E-cadherin protein and the level occurring in the membranes in both cell lines, whereas TPA caused a translocation of E-cadherin from the membrane towards the cytosol, without decreasing the total amount of E-cadherin present. The effect of both tumor promoters on connexin43 phosphorylation and localization was agent as well as cell dependent. These results show for the first time that tumor promoters can decrease the quantity and membrane localization of E-cadherin in different cell types. PMID- 8706261 TI - Re: Pereira, M.A., Barnes, L.H., Rassman, V.L., Kelloff, G.V. and Steele, V.E. (1994). Use of azoxymethane-induced foci of aberrant crypts in rat colon to identify potential cancer chemopreventive agents. Carcinogenesis, 15, 1049-1054. PMID- 8706260 TI - Persistent chemopreventive effect of S-adenosyl-L-methionine on the development of liver putative preneoplastic lesions induced by thiobenzamide in diethylnitrosamine-initiated rats. AB - S-Adenosyl-L-methionine (SAM) is a strong chemopreventive agent of rat liver carcinogenesis. Examination was made to determine whether inhibition by SAM of the development of preneoplastic liver lesions persists to SAM withdrawal in diethylnitrosamine-initiated F344 rats promoted with thiobenzamide (TB). The rats were subjected, 2 weeks after initiation, to 5 weeks feeding with a 0.1% TB diet followed by a TB-free diet for 6 weeks and then a second TB treatment for 3 weeks. SAM (384 micromol/kg/day) was injected i.m. during the first TB cycle (treatment A) or for 6 weeks after the first TB cycle (treatment B). Many gamma glutamyltranspeptidase (GGT)-positive lesions developed in initiated rats after the first TB cycle. They decreased in number after TB withdrawal, while partial recovery of lesion number and a great increase in volume occurred after the second TB cycle. Liver ornithine decarboxylase (ODC) activity and c-myc and c-Ha ras mRNAs increased during the TB cycles and returned to normal liver values after TB withdrawal. Number and size of GGT-positive lesions, DNA synthesis of GGT-positive cells, liver ODC activity and c-myc and c-Ha-ras mRNA levels decreased as a consequence of SAM treatment A. The recovery of these parameters, induced by a second TB cycle in rats not treated with SAM, was prevented by SAM treatment B. These results suggest that SAM causes a persistent decrease in growth capacity of preneoplastic liver lesions in rats subjected to a diethylnitrosamine/TB protocol. PMID- 8706262 TI - [The regulation of deep peroneal nerve inputs on depressor response evoked by caudal ventrolateral medulla excitation in rabbits]. AB - Experiments were performed on 33 white male rabbits anaesthetized with chloralose and urethane under artificial ventilation. The results were as follows: (1) depressor response could be evoked by electric stimulation of caudal ventrolateral medulla (cVLM). The inputs of deep peroneal nerve (DPN) could inhibit partly the depressor response evoked by electric stimulation of cVLM (P < 0.001). The regulation effects of DPN disappeared in 25-30 min. (2) after iv naloxone (1.5 mg/kg) or microinjection of naloxone into rostral ventrolateral medulla (rVLM) (2 micrograms/1 microliter NS), the inhibitory effect of DPN inputs was abolished partly (P < 0.01, P < 0.001). These results suggest that: (1) the depressor response elicited by cVLM excitation in rabbits can be regulated by inputs of DPN; (2) opiate peptide in rVLM may take part in the inhibitory effect of DPN inputs on depressor response excited by cVLM. PMID- 8706263 TI - [Angiotensin II increasing the inhibitory effects of electroacupuncture on the nociceptive discharge of neurons in the spinal dorsal horn]. AB - Experiments were performed on 53 rats. The nociceptive discharges of spinal dorsal horn neurons were extracellularly recorded by glass microelectrodes and tested for the effects of angiotensin II (A II) on the nociceptive discharges of spinal dorsal horn neurons and AII on the inhibitory effects of electroacupuncture (EA). The results showed as follows: (a) The nociceptive discharges of spinal dorsal horn neurons might be inhibited (25/35) or facilitated (10) with A II (50-500 ng/20 ul) applied to the surface of dorsal spinal cord (L2-3); (b) The facilitory effects (5/5) were observed with A II (2 ug/20 ul) applied to the surface of dorsal spinal cord; (c) The inhibitory effects of EA (20 Hz, 2-4 V, 5 min) could be increased with A II (250 mg/20 ul, 8/9). Therefore, A II may modulate the nociceptive response of spinal dorsal horn neurons and influence the effects of EA, and different effects can be produced when the different dosage of A II is applied to the surface of dorsal horn. PMID- 8706264 TI - [Influences of capsaicin and electroacupuncture at "point" on acetylcholine (ACh) release in the spinal dorsal horn of rats]. AB - The dynamic changes of ACh activity in the spinal dorsal horn of rats were recorded by ACh ion selective microelectrodes. The influence of applying capsaicin to the spinal surface, administering anti-SP surum to the interior of dorsal horn, cutting off dorsal roots and electroacupuncturing at "points" on the ACh activity were studied. The results showed that electroacupuncture at "points" and capsaicin could facilitate ACh release of the spinal dorsal horn, it was fully inhibited by cutting off dorsal roots or obviously inhibited by administering anti-SP surum. The above results indicated that ACh might participate in the primary afferent of painsensation and acupuncture sensation as a neurotransmitter and SP might adjust the release of ACh in spinal dorsal horn. PMID- 8706265 TI - [Influence of thoracic spinal subarachnoid microinjection of noradrenaline on the effect of electroacupuncture]. AB - Influence of thoracic spinal subarachnoid microinjection of noradrenaline (NA, 40 mg/20 microliters) on the effect of electroacupuncture (EA) at Neiguan (PC6) was observed in 40 rabbits anesthetized with urethan-chloralose. Results showed that: 1) EA of Neiguan (PC6) could significantly accelerate the recovery of ST-segment and T-wave of electrocardiogram induced by acute myocardial ischemia (AMI); 2). Subarachnoid microinjection of NA also could promote the recovery of ST, T and blood pressure of post-AMI; and 3) Subarachnoid microinjection of NA could strengthen the effect of EA of Neiguan (PC6) to promote the recovery of ST and T of Post-AMI. It suggests that alpha-receptors of intra-thoracic spinal cord participate in the action of EA of Neiguan (PC6) ameliorating AMI, and the upper thoracic segement is one of the links for connecting Neiguan and the heart. PMID- 8706266 TI - [Application of acupuncture anesthesia combining with drugs during neolarynx reconstruction]. AB - 50 Cases of neolarynx reconstruction under acupuncture anesthesia combining with drugs were performed from 1992 to 1995. We used the ear points "Lung", "Shenmen", "Sympathetic", body points "Hegu", "Zhigou", "Futu". The patients were given pethidine, rotundine, metoclopramide intramuscularly 15-25 min before operation. The excellent rate was 98%. PMID- 8706267 TI - [Effect of electroacupuncture at neiguan point on transmembrane potential of the ventricular cells of rabbits with acute myocardial ischemia in situ]. AB - Acute myocardial ischemia of rabbit was caused by Pituitrin (2.5 u/kg) intravenously. The effect of electroacupuncture (EA) at Neiguan point on transmembrane potential was observed with a suspension microelectrode technique in situ and under the condition of natural respiration; the blood pressure and electrocardiogram were recorded simultaneously. The results showed that during acute myocardial ischemia, the resting potential (RP), action potentail amplitude (APA) and maximum velocity of depolarization (Vmax) were all significantly decreased, but action potential duration (APD) was markedly prolonged and after depolarization potential appeared at the same time. All of the above mentioned changes of potentials could cause arrhythmia, while EA at Neiguan could weaken the changes of the potentials markedly, prevent the ventricular cells from changing to slow reaction cells and reduce the incidence of after depolarization potential. That may be a reason why EA at Neiguan can prevent and cure arrhythmia complicated by acute myocardial ischemia. PMID- 8706268 TI - [P-NMR analysis of the hepatic cell energetic metabolism in tumor-bearing mice by moxibustion treatment]. AB - The new technique, NMR (nuclear magnetic resonance) spectrum analysis was used in this research to measure the state of energetic metabolism of hepatic cells at the organic level. The experimental results demonstrated that moxibustion on Guanyuan (CV4) point could remarkably increase the ATP molecules in the hepatic cells, significantly raise the thermodynamic reserve and phosphate potential of the hepatic cells, so as to enhance the functional activities of liver. It is discussed here of the mechanism of anti-tumor by moxibustion treatment and of the theory of warming the kidney and enhancing the Yang to tonify to energetic Qi by moxibustion on Guanyuan point. PMID- 8706269 TI - [A study on factors of acupuncture methods affecting effects through acupuncture treatments for experimental gastric ulcer in rats]. AB - Many factors, such as the selection of acupoints, the interval between acupunctures, the acupuncture time, the manipulation and the specification of acupuncture needles, are capable of affecting curative effects. There were some studies on these factors ago. However each of these studies only related to a single factor. These studies could not reflect the clinical truth, and could not provide accurate overall data for medical staff. In order to solve the above problems, we carried out an animal experiment designed by L9 (3(4)) orthogonal design. The experiment which consisted of 9 acupuncture groups and 3 control ones was carried out in 108 rats, which were suffered from experimental gastric ulcer. In the experiment, the body weight, ulcer area, plasma SOD and LPO of each rat were observed. The results showed that in the acupuncture treatments for experimental gastric ulcer in rats, 10 minutes' electric acupuncture treatment using thinner needles and the method that each rat was treated every other day were the best methods. The reason why the above results came into being was elucidated in this paper. PMID- 8706270 TI - [The effect of moxibustion on gastric mucosa in rats and its relation to copper, zinc contents in serum]. AB - 18 rats were randomly deivided into 3 groups: control grup, the group merely with experimental gastric ulcer and the group with experimental gastric ulcer after 3 week moxibustion. The results showed: (1) moxibustion could significantly reduce the ulcer area of the rats (P < 0.05); (2) compared with control group, the experimental gastric ulcer model rats had an increase in copper content and Cu/Zu ratio, the differences were statistically significant (P < 0.05, P < 0.01); (3) in contrast to the rats merely with experimental gastric ulcer, moxibustion pretreated rats had a significant increase in Zinc content in serum (P < 0.01), but its copper content and Cu/Zn ratio in serum were significantly decreased (P < 0.05, P < 0.01). The results confirmed that moxibustion pretreatment at "Shen Jue" could protect the gastric mucosa from strass-induced ulcer, having protective effect on gastric mucosa. The results also suggested that there was a disbalance in trace elements metabolism, but moxibustion could improve it. This was most likely to be one of the mechanisms of moxibustion protection on gastric mucosa. PMID- 8706272 TI - [The effect of electro-acupuncture at auricular & body acupoints on the curve changes of the pressure-volume of urinary bladder and the electric activity of pelvic nerves of rat]. AB - The electric discharges of the post-pelvic ganglionic vesical nerve fibers were recorded to investigate the effect of acupuncture on the function of urinary bladder. It has been demonstrated that the changes of the pressure-volume of urinary bladder are controlled not only by the wall of the urinary bladder its self, but also by the regulation of parasympathetic nerve; the electro acupuncture can play an role of regulation in the functional state of urinary bladder through nerves. PMID- 8706271 TI - [Effect of electroacupuncture at "shenshu" point on renal blood flow in rabbits]. AB - The purpose of this study is to observe the effect of electroacupuncture at "Shenshu" point (U.B.-23) on the renal blood flow (RBF) under the conditions of normal, glycerol-induced renal ischemia and renal neurotomy. The RBF, which is measured by hydrogen gas clearance method, was chosen as index. The results are as follows: 1. RBF is decreased by electroacupuncturing "Shenshu" point under both conditions of normal and glycerol-induced renal ischemia. 2. After renal neurotomy, RBF is increased by electroacupuncturing "Shenshu" point. These facts suggest that the effects of electroacupuncture at "Shenshu" point relate to the renal nerve and body fluid. PMID- 8706273 TI - [Electroacupuncture promotes enlargement of adrenals and enhances level of blood corticosterone in ovariectomized rats]. AB - The present article reports EA with special acupoints may promote the enlargement of bilateral adrenals and enhance the level of blood corticosterone in ovariectomized (OV) rats. The mean of adrenal weight (M +/- SE, mg) as following, OV + EA group 71 +/- 2.1, OV group 57.4 +/- 4.7, EA group 57.6 +/- 1.1 and control group (CT group) 54 +/- 2.8 respectively. For observation of a self control, the detection of blood corticosterone contents was performed in the same animals before and after ovariectomy. The results showed that before ovariectomy the contents of blood corticosterone in the rats were 3.27 +/- 0.7 micrograms/100ml before EA and 3.63 +/- 0.9 micrograms/100ml after EA. No significant difference of blood corticosterone level was found before and after EA. After the animals ovariectomized the level of blood corticosterone increased from 4.38 +/- 0.62 micrograms/100ml to 6.02 +/- 0.77 micrograms/100ml following EA treatment (P < 0.01). No similar effect of EA treatment with control acupoints in OV rats was seen. These results suggest that EA may regulate the abnormal function of endocrine and this effect of EA possesses relative specificity of acupoint. PMID- 8706274 TI - [Effect of HC-3 on electroacupuncture-induced immunoregulation]. AB - Our previous study suggested that peripheral sympathetic nerve played an important role in the electroacupuncture (EA)-induced immunomodulation. The aim of this study is to explore the role of peripheral parasympathetic nerve in it. All mice are preimmunized with SRBC (sheep red blood cell) 4 days before experiment. The animals in different groups were injected i.p. with Hemicholine-3 (HC-3, an ACh synthesis blocker) or physiological saline 3 hr before EA. The main results are as follows: 1. In saline group, the LTT (lymphocyte transformation test), IL-2 (interleukin-2) of spleen and lymphocyte number of thymus are markedly increased, and the other immune parameters have no change after EA. 2. In HC-3 control group, the LTT, IL-2 are decreased significantly and the other immune parameters have not shown influence. 3. In the HC-3 with EA group, the magnitude of LTT, IL-2 did not significantly change as compared with HC-3 alone, it is especially noteworthy that the other unchanged parameters by HC-3 such as spleen weight, spleen index, thymus weight, thymus index, number of spleen cells, spleen LZM(lysozyme) and thymus LZM are clearly decreased after EA. The above results suggest that peripheral parasympathetic nerve may play some promoting effect in the EA-induced immunomodulation. Its effect may be mediated by ACh released from parasympathetic nerve endings. PMID- 8706275 TI - [Influence of changing Ca++ concentration in neiguan (PC 6) on the effect of acupuncture treating experimental arrhythmia of rabbits]. AB - We have made two kinds of experimental arrhythmia of rabbits by injecting aconitine and stimulating hypothalamus. Acupuncture could improve arrhythmia, after Ca++ in Neiguan (PC6) was chelated with EDTA solution the effect of acupuncturing Neiguan (PC6) was abolished. It indicates that Ca++ may be the key factor of acupuncture effect and one of the important material bases of the functional activity of meridians and collaterals. PMID- 8706276 TI - [Studies on regulatory effects of acupuncture on mucosal secretory IgA in patients with allergic asthma]. AB - We studied mucosal SIgA immunity of patients with allergic asthma treated by acupuncture. Our results showed that concentrations of SIgA and total IgA in saliva (P < 0.01, < 0.02) and in nasal secretions (P < 0.02, < 0.02), and levels of IgE in sera (P < 0.001) from patients with allergic asthma were significantly decreased after treatment with acupuncture. The results suggested that attacks of delayed and immediate allergic asthma could be effectively inhibited by acupuncture. The facts that there were no significant changes in IgA in supernatants of peripheral blood lymphocytes stimulated with PWM of patients with allergic asthma after treated by acupuncture were associated with regulation of negative feedback of T-lymphocyte expressing IgA-R and it released bound factor of IgA in production of polymeric IgA. PMID- 8706277 TI - [Effect of electrical stimulation of red nucleus on acupuncture analgesia]. AB - The function of red nucleus (RN) was important in motor control. This work was to study whether the RN influenced the effect of EA and the somatosensory afferent system. C-responses of spinal dorsal horn neurons (SDHN) were recorded as nociceptive responses. Electrical stimulation of RN could intensify the inhibitory effect of EA and inhibit nociptive response of SDHN. Naloxone (NX) could completely block that inhibition of RN. The result inferred that the RN can modulate both of somatomotion and somatosense. The endogenous opiate system is involved in the somatosensory modulation of RN. PMID- 8706278 TI - [A study of relation between rheumatoid arthritis (RA) and blood stasis--the effect of acupuncture promoting blood circulation to remove blood stasis]. AB - In this article, the clinical 31 cases patients with Rheumatoid artritis (RA) were observed and animal experiments were carried out on RA model rats and the treatment of acupuncture with combination of the different points was adopted. It was found that most of the 31 cases of RA patients had the signs of bloods stasis and abnormal hemorheology, statistical analysis indicated that there were significant differents between RA patients and healthy person. Red and swelling in joints of four limbs, ear and tail, clubbed toes, abnormal hemorheology, high in blood platelet aggregation and so on were found in model rats of RA, statistically, sinificant differents also exist between model rats of RA and healthy rats. The above indesexes in patients and model rats of RA were significantly improved after acupuncture treatment, which suggested that (1) there be a close relation between RA and blood stasis. (2) acupuncture play a role of promoting blood circulation to remove blood statis. (3) examination of hemorheology may be one of standards that development of RA and curative effect. PMID- 8706279 TI - [Confocal laser scan microscope system and its applications on studying acupuncture and meridian]. AB - In this paper, the principle of a confocal laser scan microscope system and its advantages are introduced. Its applications in biology and research of acupuncture and meridian are also introduced. Compared with conventional microscope, the confocal laser scan microscope system has many advantages such as high resolution fluorescent image, 3-dimension reconstruction, images quantitative analysis of concentration of ion within a cell, cell-cell communication and so on. It will offer many mathods for researching acupuncture and meridian. PMID- 8706280 TI - Assessment of cure after transsphenoidal surgery for Cushing's disease. AB - The best long-term results of transsphenoidal surgery appear to be in those patients who have unmeasurable serum cortisol levels in the immediate post operative period. This appears to be due to isolated ACTH deficiency in the remaining normal pituitary gland. In our experience, however, long-term clinical remission may be possible with measurable serum cortisol levels post-operatively. The cortisol response to ACTH or CRF appears to provide no greater discrimination than basal measurements and is possibly less helpful than dexamethasone suppression. The prolonged clinical remission that we have seen in some patients with early measurable but low cortisol levels, together with the possible operative morbidity and partial success of repeat surgery, suggests careful assessment before a recommendation is made of an immediate second operation in these patients. The possibility of cyclical secretion of cortisol must be considered both pre and post-operatively. Finally, as experience with transsphenoidal surgery is still relatively short, all these patients need continuing endocrine follow-up in centres with adequate experience of the condition. For the individual patient we recommend that decisions regarding further therapy are based on 0800-0900 h serum cortisol levels, an index of cortisol production (serum cortisol day profiles or 24 hour urinary free cortisol), and the response to low dose dexamethasone suppression in the early post-operative period. PMID- 8706281 TI - Evidence for the Bauman variant in Kallmann's syndrome. AB - BACKGROUND AND OBJECTIVE: Idiopathic hypogonadotrophic hypogonadism (IHH) is a condition of gonadotrophin releasing hormone (GnRH) deficiency. IHH associated with anosmia is Kallmann's syndrome. A variant has been described by Bauman where a patient with Kallmann's syndrome apparently regained normal hypothalamo pituitary function 2 years after the initial diagnosis. GnRH secretory activity can be assessed by measuring LH pulsatility. Our objective was to define the pattern of LH pulsatility in men with IHH and Kallmann's syndrome compared with those of normal controls, and to determine whether there is evidence for a Bauman variant of Kallmann's syndrome. DESIGN: Patients with IHH and Kallmann's syndrome were recruited from the endocrine clinic. Long-term hormone replacement therapy was discontinued. LH pulsatility was determined. PATIENTS: Three men with IHH, 3 men with classical Kallmann's syndrome and 5 normal male volunteers. MEASUREMENTS: Baseline serum FSH, LH and testosterone. Intensive blood sampling every 10 minutes for serum LH from 1000 to 1600 h during the day and 2200 to 0400 h during the night to measure LH pulsatility. RESULTS: The volunteer group showed normal LH pulsatility. In the patient group, LH secretion was apulsatile in one, showed significantly diminished amplitude in four, and there was normal pulsatility in one patient which remained normal 5 months later. CONCLUSION: Three patients with idiopathic hypogonadotrophic hypogonadism and 2 with Kallmann's syndrome had variable degrees of GnRH deficiency. One patient with Kallmann's syndrome had apparently normal GnRH activity, which remained normal 5 months later. This patient appears to have the Bauman variant of Kallmann's syndrome. PMID- 8706282 TI - A case of silent 21-hydroxylase deficiency with persistent adrenal insufficiency after removal of an adrenal incidentaloma. AB - A case of an adrenal incidentaloma in a 57-year-old man with silent 21 hydroxylase deficiency is reported. Abdominal computed tomography revealed a right adrenal tumour of 6cm in diameter. There was no evidence of adrenal hormone excess. However, after surgical removal of the adrenal tumour, the patient developed acute adrenal insufficiency. Adrenocortical function has remained low ever since surgery. Pathological examination of the tumour revealed a cortical adenoma. On the basis of increased plasma renin activity and serum 17 alpha hydroxyprogesterone concentration and decreased 21-hydroxylase activity of the adenoma tissue, the patient was diagnosed as having systemic 21-hydroxylase deficiency. There are two possible mechanisms for the persistent adrenal insufficiency; first the residual left adrenal gland may have been originally hypo-functioning for some reason and, second, the left gland may have lost the ability to regenerate following prolonged suppression by the adenoma. In cases of adrenal incidentalomas with 21-hydroxylase deficiency, the indications for surgical removal should be carefully considered. PMID- 8706283 TI - Growth hormone deficiency and hypogonadism in a patient with multiple sclerosis. AB - We describe the case of a 30-year-old female patient with a 7-year history of multiple sclerosis, who presented with an 18-month history of secondary amenorrhoea and vague symptoms which included poor sleep and impaired concentration. Endocrine investigations revealed hypogonadotrophic hypogonadism and GH deficiency, a probable consequence of a hypothalamic plaque. This is the first report of hypogonadotrophic hypogonadism and GH deficiency occurring in conjunction with multiple sclerosis. As such, it should raise suspicion of endocrine dysfunction occurring in a condition with such a vast spectrum of disability as multiple sclerosis. PMID- 8706284 TI - Pituitary apoplexy after leuprolide administration for carcinoma of the prostate. AB - The syndrome of pituitary apoplexy has been reported to occur after the administration of several different medications. We report a case in which pituitary apoplexy developed shortly after the administration of leuprolide in a patient with prostate cancer. Leuprolide is a potent gonadotrophin releasing hormone (GnRH) analogue used to suppress leuteotrophic hormone (LH) and testosterone levels in patients with metastatic prostate cancer. LH and testosterone levels actually rise in the first week after its administration before becoming suppressed. We suspect that this acute stimulating effect of leuprolide is linked to the acute onset of pituitary apoplexy in a patient with a possible gonadotrophoma. PMID- 8706285 TI - Clinical and genetic changes in a case of a Cushing's carcinoma. PMID- 8706286 TI - Percutaneous ethanol injection for thyroid cysts: a word of caution. PMID- 8706287 TI - Screening for Down's syndrome. PMID- 8706288 TI - Second trimester screening for Down's syndrome using maternal serum dimeric inhibin A. AB - BACKGROUND AND OBJECTIVE: Prenatal maternal serum screening for Down's syndrome has become an important and established part of modern antenatal care. Previously it has been reported that non-specific immunoreactive inhibin may be useful in this context. Using a novel assay we have evaluated dimeric inhibin A as a possible second trimester marker of Down's syndrome. METHODS: From 1992-1993 records, stored sera from women with Down's affected pregnancies and chromosomally normal control pregnancies were identified and retrieved for analysis. These sera had been prospectively collected at 15, 16 and 17 weeks gestation. SUBJECTS: Records revealed 21 women who had had a Down's syndrome pregnancy and who also had serum available for analysis. Sera from 150 chromosomally normal controls, matched for gestation and duration of storage, were also retrieved. MEASUREMENTS: Dimeric inhibin A was measured using a recently developed two-site enzyme-linked immunoassay. This employs a capture anti inhibin beta A-subunit monoclonal antibody, covalently bound to a microtitre plate and a second anti inhibin alpha-subunit antibody conjugated to alkaline phosphatase, allowing detection. RESULTS: The mean (95% CI) maternal serum dimeric inhibin A in the samples from control pregnancies was 237 (201.5-273.4) ng/l, 266.9 (235.4-298.5) ng/l and 207.2 (178.5-235.9) ng/l at 15, 16 and 17 weeks gestation respectively. Expressing the results from the Down's samples as multiples of the normal median (MoM), the median (95% CI) MoM was 2.6 (2.25 3.57), significantly higher than the controls (P < 0.0001, Mann-Whitney U-test). In the sample set tested, for a given false positive rate of 5.3% inhibin A alone afforded a detection rate of 62%, detecting cases previously undetected by routine screening. CONCLUSIONS: Dimeric inhibin A appears to be a promising new marker for the prenatal detection of Down's syndrome. Further prospective evaluation and assessment with other established markers would now be merited. PMID- 8706289 TI - Continuous subcutaneous infusion of low dose growth hormone decreases serum sex hormone binding globulin and testosterone concentrations in moderately obese middle-aged men. AB - OBJECTIVE: Sex-hormone binding globulin (SHBG) is a liver derived protein whose concentration has been shown to be affected by a number of factors. The aim of the present study was to investigate the possible effect of increased basal GH concentrations on serum concentrations of SHBG, testosterone and thyroid hormones. DESIGNS: Recombinant human growth hormone (rhGH) was given as a continuous subcutaneous infusion in a low dose (0.02 U/kg/day) over a period of 14 days in an open study. PATIENTS: Eight middle-aged (42-59 years) overweight (body mass index 26.1-33.8 kg/m2) but otherwise healthy men were studied. MEASUREMENTS: Blood samples were obtained after an over-night fast before and after 2, 7 and 14 days of treatment. Serum was separated and stored at -20 degrees C until assay. RESULTS: Serum GH concentrations increased to a steady level of 2-4 mU/l. Serum SHBG concentration decreased between 2 and 7 days of treatment and serum testosterone concentration changed in parallel. There was no change in the ratio between serum SHBG and serum testosterone. Serum T4 and free T4 concentrations decreased, and that of T3 increased as a result of continuous GH infusion. Simple correlations between changes in SHBG concentrations and other hormonal changes showed positive significant correlations between changes of SHBG and of testosterone. CONCLUSIONS: Low dose continuously infused GH resulted in a parallel decrease in serum SHBG and testosterone concentrations. Thyroid hormone concentrations were affected in a similar way to that previously demonstrated following daily injections of GH. PMID- 8706291 TI - The effect of oophorectomy and hormone replacement on neurohypophyseal hormone secretion in women. AB - OBJECTIVE: There is evidence to suggest that the release of neurohypophyseal hormones may be influenced by the circulating concentrations of gonadal steroids. We therefore monitored this relationship in women undergoing prophylactic bilateral oophorectomy at the time of hysterectomy with subsequent hormone replacement therapy and compared it with that in women undergoing hysterectomy with conservation of ovaries. DESIGN: Patients were randomly allocated to receive either transdermal oestradiol patches, 0.05 mg/day, or subdermal implants containing either 50 mg oestradiol or 50 mg oestradiol with 100 mg testosterone. Blood samples for determination of plasma hormone concentrations, electrolytes and osmolality were obtained immediately before and after surgery and then at two monthly intervals for 8 months and finally at 12 months. MEASUREMENTS: Free oestradiol, vasopressin and oxytocin were measured by radioimmunoassay. RESULTS: Vasopressin concentrations were found to fall after surgery in oophorectomized women, but not in those with ovaries. There were no changes in fluid balance to account for the reduced plasma vasopressin concentrations. During treatment oestradiol appeared to enhance and testosterone to suppress vasopressin release. Oophorectomy had no significant effect on plasma oxytocin concentrations, but in the groups receiving oestradiol implants concentrations fell significantly at 8 and 12 months compared with the value at 6 days. CONCLUSION: The observed changes in plasma vasopressin concentrations were consistent with the observations in experimental animals and provide evidence that vasopressin release in the human is influenced by gonadal steroids. PMID- 8706290 TI - Oropharyngeal regulation of water balance in polydipsic schizophrenics. AB - OBJECTIVE: Disordered water balance causes substantial morbidity in a subset of schizophrenics and is the consequence of unexplained defects in the regulation of fluid intake and antidiuretic function. We aimed to determine whether oropharyngeal regulation of water balance is altered in these patients. DESIGN: A 2-hour infusion of 3% saline, followed 35 minute later by an oral water load (10 ml/kg over 3 minutes). PATIENTS: Age and sex-matched polydipsic schizophrenics with (n = 5) and without (n = 8) hyponatraemia; non-polydipsic schizophrenics (n = 6); and normal controls (n = 13). MEASUREMENTS: Plasma osmolality, sodium, AVP, and reported desire for water (expressed in cups), determined prior to, and for 30 minutes following, the water load. RESULTS: Plasma osmolality and sodium were consistently lower in the hyponatraemics (P < 0.01). Significant changes did not occur until 20 and 25 minutes, respectively, after oral water loading, and were similar across the four groups. AVP levels were consistently lower in the two polydipsic groups (P < 0.001), fell within 5 minutes after drinking and then levelled off. Neither the acute fall nor the overall pattern of the responses differed across groups. Subjective desire for water also decreased within 5 minutes of drinking, and also to a similar extent, in the four groups. Subsequent levels remained suppressed in the non-polydipsic groups, but rebounded toward baseline in the two polydipsic groups. Thus the overall patterns differed (P < 0.05). At the end of the study, ad lib intake correlated significantly with reported desire for water (r = 0.51, P < 0.002). CONCLUSIONS: The oropharyngeal regulation of water intake is disrupted in polydipsic schizophrenics with and without hyponatraemia. In contrast, the oropharyngeal regulation of AVP secretion appears preserved in the hyponatraemic subset. Previously observed elevations in plasma AVP in this subset are thus unlikely to be related to defects in oropharyngeal regulation. PMID- 8706292 TI - A novel mutation in the coding region for neurophysin-II is associated with autosomal dominant neurohypophyseal diabetes insipidus. AB - OBJECTIVE: Autosomal dominant neurohypophyseal diabetes insipidus (ADNDI) is a rare cause of diabetes insipidus, in which AVP serum levels are insufficient. AVP is synthesized along with neurophysin-II (NPII) as an AVP-NPII precursor polypeptide in the hypothalamus. After proteolytic cleavage during axonal transport, AVP and NPII are reassembled and stored loosely bound to each other in the posterior pituitary until both are released into the circulation. In this study, we investigated the genetic basis of ADNDI in a German kindred with 10 affected members spanning three generations. DESIGN: Genomic DNA was isolated from peripheral blood leucocytes. The entire coding region of the AVP-NPII gene of one of the affected persons was amplified by polymerase chain reaction (PCR) and subjected to nucleotide sequence analysis. Sequencing results were confirmed by restriction enzyme analysis of PCR products. PATIENTS: Six affected and two unaffected members of a family with ADNDI and 54 unrelated healthy control subjects were studied. RESULTS: The index patient was found by direct sequencing to be heterozygous for a G to T transversion at nucleotide position 1884 (exon 2) of the AVP-NPII gene. This mutation introduced a new recognition site for the restriction enzyme Ava II, which was used to test for the presence of the mutation in other family members and in control subjects. The mutation was detected in all family members with ADNDI, but was not found in unaffected family members or in control subjects. The mutation encodes a valine in place of the normal glycine at amino acid 65 of NPII, which is known to be highly conserved during evolution. CONCLUSIONS: In this family, the autosomal dominant neurohypophyseal diabetes insipidus phenotype cosegregates with a point mutation in a region of the AVP-neurophysin-II gene which codes for the carboxy-terminal domain of neurophysin-II. Although the altered amino acid is not directly involved in AVP binding, the mutation might lead to conformational changes that impair the dimerization of neurophysin-II molecules. This could in turn affect the AVP binding affinity of neurophysin-II or might interfere with the transport of the AVP-neurophysin-II precursor in the AVP-producing cells of the hypothalamus. PMID- 8706293 TI - The relation between thyroid function and nutritional status in HIV-infected patients. AB - OBJECTIVE: The effects of human immunodeficiency virus (HIV) infection on thyroid function have been reported in only a few studies with discrepant results. The aim of this study was to assess the relation between nutritional status and thyroid function in HIV infected patients. DESIGN: Prospective, cross-sectional study. SETTING: A 500-bed teaching and referral hospital serving a population of 450,000. PATIENTS: Seventy-five consecutive HIV infected patients between 21 and 40 years of age (mean 31.8 +/- 0.9 years). MEASUREMENTS: Nutritional status was evaluated using the body mass index (BMI), triceps skinfold thickness (TSF), mid arm muscle circumference (MAMC), and serum albumin concentration (SA). Hormone assays for serum T4, free thyroxine index (FTI), T3, reverse triiodothyronine (rT3), thyroxine-binding globulin (TBG), TSH and simultaneous CD4 lymphocyte counts were determined in all patients. RESULTS: Clinical stage was significantly related to nutritional status (P = 0.0001 for BMI, P = 0.0002 MAMC). The more poorly nourished groups had low mean serum T3 and rT3 levels, particularly for muscular (P = 0.0001 for T3 and P = 0.0076 for rT3) and visceral (P = 0.00001 for T3 and P = 0.0021 for rT3) protein compartments. Multivariate analysis showed that two factors, SA and MAMC, correlated significantly and independently with serum T3 and rT3. CONCLUSIONS: A close relation exists between serum thyroid hormone levels and nutritional status in HIV infected patients. These patients are probably euthyroid and the abnormal findings in the thyroid function tests are thus a reflection of the severity of illness. PMID- 8706294 TI - beta-cell function and glucose and lipid oxidation in Graves' disease. AB - OBJECTIVE: Abnormal glucose metabolism with impaired glucose tolerance has been documented in patients with thyrotoxicosis but the pathogenesis is not fully understood. Therefore, the aim of the present study was to study the beta-cell function and the meal induced oxidative glucose and lipid metabolism in patients with thyrotoxicosis. DESIGN: After an overnight fast the impact of hyperthyroidism on standard mixed meal induced glucose oxidation, lipid oxidation and beta-cell function was studied. PATIENTS: Nine untreated patients with Graves' disease were compared to 9 age and weight matched healthy controls. MEASUREMENTS: Glucose and lipid oxidation were studied by indirect calorimetry before and after the meal. The insulin secretion rate was calculated by the 'combined model' approach, after which the insulin secretion rates and the ambient glucose levels were cross-correlated. The slope of these regression lines was used as a measure of beta-cell sensitivity to glucose and denotes the insulin secretory capacity. beta-Cell function was further evaluated by measurement of proinsulin and its conversion intermediates. Glucoregulatory hormones were also measured. The findings were correlated to the thyroid hormone levels. RESULTS: Fasting blood glucose and post-prandial glucose response were increased in patients (P < 0.01). The hyperthyroid patients displayed a 'dual' beta-cell defect: (a) inability to increase the insulin response appropriately to hyperglycaemia and (b) increased proinsulin levels both in the fasting state and in response to a meal. Indirect calorimetry showed increased lipid oxidation in the fasting state and at the end of the meal (P < 0.01). No difference in glucose oxidation was demonstrated in the fasting state but the post-prandial glucose oxidation was enhanced in the patients (P < 0.01). The adrenaline response was normal, whereas the noradrenaline response was impaired or absent in the patients. The thyroid hormone levels were significantly correlated to fasting levels of blood glucose, insulin, free fatty acids and lipid oxidation, but not to fasting C-peptide, glucose oxidation or catecholamines. CONCLUSIONS: Untreated Graves' disease was associated with glucose intolerance due to quantitative as well as qualitative beta-cell defects. The lipid oxidation was increased in the fasting state and at the end of the meal; after the meal the increase in glucose oxidation was more pronounced in the patients. Thyroid hormones thus increased the oxidation but not by an increase in catecholamines. Indeed, the post-prandial sympathetic response was blunted. PMID- 8706295 TI - Hexarelin, a synthetic growth hormone releasing peptide, stimulates prolactin secretion in acromegalic but not in hyperprolactinaemic patients. AB - OBJECTIVE: In man, new synthetic peptides such as hexarelin have been shown to have a potent and dose dependent GH releasing activity. Furthermore, a significant PRL releasing activity has also been demonstrated, but this has been investigated in less detail. We have therefore evaluated the effect of hexarelin on PRL and GH secretion in patients with active acromegaly or pathological hyperprolactinaemia. DESIGN: Hexarelin (2 micrograms/kg i.v.), a modified derivative of GHRP-6 of the following structure: His-2-Me-D-Trp-Ala-Trp-D-Phe-Lys NH2, or placebo, was administered in random order on two separate occasions. PATIENTS: Eight patients with active acromegaly (ACRO, 6 F and 2 M, mean age 61.7 years, range 56-73), 6 with macroadenomas and 2 without radiological signs of tumour, and 6 female patients with pathological hyperprolactinaemia (HPRL, mean age 31.2 years, range 18-47) 5 with microadenomas and 1 with empty sella, were studied. Fourteen normal subjects (NS, 8 F and 6 M, 27.1 years, 24-30) were studied as controls. MEASUREMENTS: GH and PRL levels were evaluated every 15 minutes for 2 hours after hexarelin or placebo. Both hormones were measured using commercial IRMA kits. Basal IGF-I was measured in all subjects using an RIA following acid-ethanol extraction. RESULTS: Hexarelin induced a significant increase in PRL levels in NS (median, range, delta peak HEX vs placebo: 150 (-14 402) vs 10 (-34-24) mU/l; delta AUC HEX vs placebo: 7710 (2,100-3 2540) vs 30 ( 2,566-2,040) mU min/l, P < 0.01) and in ACRO (190 (-10-496) vs 6 (-100-34) mU/l; 10,170 (-5,310-51,436) vs -82 (-6,030-1,410) mU min/l, P < 0.02), but not in HPRL (10 (-180-80) vs 50 (-100-240) mU/l; -600 (-16,996-10,140) vs -1,950 (-8,540 14,160) mU min/l). Hexarelin also induced a lower increase of GH in HPRL (60 (30 82) vs 1.8 (-0.2-2.2) mU/l; 2,853 (1,477.6-4,372.6 vs 91.6 (-160.6-174) mU min/l, P < 0.05) than in NS (90.8 (50.6-181) vs 0.8 (-1.2-6.8) mU/l; 6642 (2,004 13,252.6 vs 42 (456-900) mU min/l, P < 0.01) or in ACRO (117.2 (21.2-420.6 vs 3.8 (-2.2-18) mU/l; 6645 (1,554-22,138.6 vs 334.6 (-324-1,065) mU min/l, P < 0.02). CONCLUSIONS: Our data show that the PRL releasing effect of hexarelin is preserved in acromegaly but lost in pathological hyperprolactinaemia. In contrast with acromegaly, the GH releasing effect of hexarelin is also blunted in hyperprolactinaemic patients. These data demonstrate that patients with pathological hyperprolactinaemia are partially refractory to the activity of hexarelin. PMID- 8706296 TI - The effect of beta-endorphin on basal and insulin-hypoglycaemia stimulated levels of hypothalamic-pituitary-adrenal axis hormones in normal human subjects. AB - OBJECTIVE: It has been demonstrated that beta-endorphin reduces CRH production and hypoglycaemia-induced ACTH secretion in the rat. We aimed to determine whether supraphysiological levels of beta-endorphin inhibit the ACTH and CRH response to insulin-induced hypoglycaemia in human subjects. DESIGN: Plasma glucose, prolactin, cortisol, ACTH, CRH and AVP were measured at intervals over a 3-hour period. Intravenous beta-endorphin 5 mg/50 ml or an equal volume of normal saline was infused between 30 and 90 minutes, with soluble insulin 0.15 units/kg administered i.v. at 60 minutes in a cross-over design. SUBJECTS: Six healthy male volunteers aged 20-35 years. MEASUREMENTS: Prolactin was measured by a fluoroimmunometric assay, ACTH, CRH and AVP by radioimmunoassay, and cortisol was measured by enzyme-linked immunosorbent assay. Haemodynamic measurements were recorded prior to each blood sample. Results are expressed as mean +/- standard error of the mean. RESULTS: beta-Endorphin resulted in a significant decrease in baseline cortisol (P < 0.05) but not ACTH. Plasma glucose (P < 0.001) and CRH (P < 0.05) and PRL (P < 0.05) increased significantly during beta-endorphin compared to normal saline. After insulin administration, glucose reached a similar nadir during beta-endorphin and normal saline (2.1 +/- 0.1 and 1.9 +/- 0.15 mmol/l, respectively) but the fall in plasma glucose was delayed during beta-endorphin (P < 0.01 by ANOVA). This resulted in a significantly altered time-course for the ACTH and cortisol responses (P < 0.05 for each), but no difference overall in the magnitude of the response. In contrast, neither the timing nor the magnitude of the CRH and AVP responses were affected. Prolactin also reached a similar peak value after the administration of insulin, while the haemodynamic responses to hypoglycaemia were not significantly altered during beta-endorphin. CONCLUSIONS: While beta-endorphin has been shown to be inhibitory to basal ACTH and cortisol secretion in humans, we note a significant increase in plasma CRH in response to beta-endorphin, which may be arising from a peripheral source. Intravenous beta endorphin increases plasma glucose and delays the onset of hypoglycaemia following insulin but does not result in significant inhibition of the ACTH and cortisol response. This may reflect the poor penetration of beta-endorphin into the central nervous system, although a hypothalamic effect of beta-endorphin is implied by the increased PRL. The significantly delayed time course in ACTH and cortisol secretion noted during beta-endorphin is not explained by a later response of either CRH or AVP. Although peripheral levels of these hormones may be a relatively insensitive measure of hypothalamic function, an additional factor may influence ACTH release during hypoglycaemia. PMID- 8706297 TI - Association of HLA-DQA1*0501 with Graves' disease in English Caucasian men and women. AB - OBJECTIVE: A recent report has demonstrated a stronger association between the HLA-DQA1*0501 allele and Graves' disease in Caucasian men than in women. Our aim was to confirm this association in a larger series of male Caucasian patients. DESIGN: Polymerase chain reaction/sequence specific oligonucleotide probing (PCR/SSOP). PATIENTS: Fifty men and 70 women with Graves' disease were studied as well as a control group consisting of 57 healthy, unrelated men and women. METHODS: Genomic DNA was derived from venous blood samples. Appropriately primed DNA was amplified by PCR and the products were subjected to SSOP. The presence of the allele was demonstrated by enhanced chemiluminescence. RESULTS: A significant association between HLA-DQA1*0501 and Graves' disease was demonstrated among both the men and women as well as in the combined disease cohort, with HLA-DQA1*0501 conferring a greater relative risk than HLA-DR3 in all three groups. This association persisted when the results from the DR3-negative Graves' patients were analysed in isolation. HLA-DQA1*0501 was heterogeneously distributed between the sexes with significantly more female Graves' patients carrying this allele. CONCLUSIONS: There is a significant association between HLA-DQA1*0501 and Graves' disease which may be unrelated to the inheritance of this allele on an extended haplotype with HLA-DR3. In contrast to a recent report, HLA-DQA1*0501 was significantly more prevalent among women with Graves' disease than men. PMID- 8706298 TI - Low and conventional dose transdermal oestradiol are equally effective at preventing bone loss in spine and femur at all post-menopausal ages. AB - OBJECTIVES: We wished to appraise the effectiveness of hormone replacement therapy (HRT) in early (< 67 years) and late (> 67 years) post-menopausal women referred to a metabolic outpatient clinic for assessment of their bone status. Because older women often experience side-effects with conventional HRT, a low dose preparation (Estraderm 25) was also compared with conventional HRT (Estraderm 50). DESIGN AND SETTING: Since all patients were symptomatic, the investigation was open and not placebo controlled. Patients were offered HRT and told about the two dosages. If they wished to use HRT, allocation of dosage was made randomly unless there were reasons to use a specific dose. PATIENTS: One hundred and ninety-six women were studied over 1 or 2 years with 80 reaching 3 years of treatment. Patients were divided into those under 67 years and those over 67 years at the start of treatment. Each group was further divided into those taking Estraderm 25 and those taking Estraderm 50 with norethisterone if appropriate. MEASUREMENTS: Bone mineral density (BMD) was measured (DXA, Hologic) at the lumbar spine and femoral neck at 0 year (196 patients), at 1 year (169 patients), at 2 years (139 patients) and at 3 years (80 patients). Patients losing bone were expressed as those whose 3 year BMD was lower than initial or as those whose BMD at 3 years had fallen by more than twice the coefficient of variation for that site (non-responders). RESULTS: In lumbar spine, BMD increased maximally in the first year in all groups and the gain was maintained after 3 years. The change was similar whether patients were divided by age or dosage. For those on Estraderm 25, mean change after 3 years was 8.1 +/- 6.8% and on Estraderm 50, 9.0 +/- 8.3% (combined 8.7 +/- 7.8%). Only 3.9% of patients were non-responders at the lumbar spine after 3 years. At femoral neck, changes were significant at 3 years only in the Estraderm 25 > 67 years and Estraderm 50 < 67 years groups and averaged 2.3 +/- 5.4% for all patients. At the femoral neck, 10.4% of patients were non-responders after 3 years. Percentage change of BMD over 3 years at lumbar spine correlated with that at the femoral neck (r = 0.56). Percentage change of BMD at lumbar spine over 3 years correlated with menopausal age (r = 0.295). No relation was found between dosage of Estraderm/kg body weight and response of BMD at either site. CONCLUSIONS: Transdermal oestrogen is effective at preventing bone loss in the spine at all post-menopausal ages and is capable of doing this in low dosage. Prevention of bone loss at the femoral neck is less certain and the average change in BMD over 3 years was significantly lower (P < 0.001) than in the lumbar spine. Use of Estraderm 50 is not associated with a greater response of bone mass and there was no evidence of an increasing BMD response as oestradiol dosage/kg body weight increased. PMID- 8706299 TI - Polymorphism of glycogen synthetase gene in polycystic ovary syndrome. AB - OBJECTIVE: Polycystic ovary syndrome is a heterogeneous disorder associated with a moderate degree of insulin resistance and a higher risk of developing NIDDM. The exact mechanism of insulin resistance is unclear. This study examines the frequency of an Xbal polymorphism of the glycogen synthetase gene (A2 allele) as a marker of insulin resistance and seeks to relate the presence of the A2 allele to indices of insulin sensitivity in women with polycystic ovary syndrome (PCOS). METHODS: Insulin sensitivity was assessed by fasting insulin measurements, as well as following oral glucose tolerance test. An i.v. insulin tolerance test was performed to measure the rate of endogenous blood glucose disposal following an i.v. bolus of insulin. Restriction fragment length polymorphism was performed with Xbal digestion of PCR amplified product to detect the presence of A1 and A2 allele. PATIENTS: Seventy-one obese (BMI > 25.1) and 19 non-obese (BMI < 25) women with PCOS, and 62 controls (33 obese and 29 non-obese) participated in the study. RESULTS: Obese PCOS had significantly higher fasting insulin (P = 0.002) compared to obese controls. There was no difference between non-obese PCOS and controls. Twenty per cent of obese PCOS had impaired glucose tolerance. The A1A2 genotype was detected in 16 of the 150 (10.7%) subjects examined. Of these, 11/88 (12.5%) were PCOS and 5/62 (8%) were controls. The A2A2 genotype was not present in any of the subjects. The A1A2 genotype was not detected in any of the subjects with impaired glucose tolerance. There was no significant difference in the incidence of the A1A2 genotype between PCOS and controls or between the individual groups. There was no association between the presence of the A1A2 genotype and indices of insulin sensitivity. CONCLUSION: The Xbal polymorphism (A2 allele) of the glycogen synthetase gene was not over represented in the PCOS subject and did not relate to the indices of insulin sensitivity or glucose intolerance. PMID- 8706300 TI - The differential regulation of the circulating levels of the insulin-like growth factors and their binding proteins (IGFBP) 1, 2 and 3 after elective abdominal surgery. AB - OBJECTIVES: Patients undergoing abdominal surgery often suffer from morbidity associated with increased protein catabolism. Therapeutic recombinant human insulin-like growth factor (rhIGF)-I has been proposed as a means of reversing this process. As IGFBPs modulate the bioavailability of the IGFs, we have studied the changes in the circulating levels of these peptides during surgery. DESIGN: Patients undergoing elective intestinal surgery were recruited prospectively. Blood samples were taken before, during and after surgery. Standard anaesthetic techniques were used. METHODS: Twelve adults (aged 30-70 years; 9 female, 3 male) undergoing surgery were studied. Serum was taken before premedication (preop), end of surgery (end surg), 2 h, 6 h post surgery, on days 1-4, 7, 10 and 14, and on recovery at 6 weeks. MEASUREMENTS: Serum IGF-I, IGF-II, IGFBP-1, IGFBP-2, IGFBP-3, insulin and C-peptide were measured by radioimmunoassay. IGFBP profiles were also assessed by Western ligand blot (WLB). Samples taken preop and at 2 days were separated by fast-phase liquid chromatography (FPLC) using a Superose 12 column under neutral conditions (pH 7.4), and the fractions were analysed subsequently by WLB and immunoblot using a specific IGFBP-3 antiserum. RESULTS: IGF-I fell rapidly during surgery from 170 +/- 21 (preop) to 133 +/- 14 micrograms/l (end surg) (P < 0.05). The magnitude of this fall could not be explained by haemodilution. IGF-I levels then fell further to a nadir of 103 +/- 10 micrograms/l at day 4 (P < 0.05). IGF-II fell from 580 +/- 46 (preop) to 397 +/- 38 micrograms/l (day 2). Both IGF-I and IGF-II recovered to preop levels at 6 weeks (205 +/- 14 micrograms/l and 623 +/- 30 micrograms/l respectively). IGFBP-3 levels fell similarly from 4.46 +/- 0.45 to 3.2 +/- 0.3 mg/l (end surg) and to a nadir of 2.66 +/- 0.19 mg/l at day 2. There was a close correlation between IGFBP 3 levels and the sum of IGF-I and IGF-II levels before surgery (r = 0.9, P < 0.01) and this was maintained throughout the post-operative period (mean correlation coefficient of 0.86 +/- 0.02, P < 0.05). On days 2 and 3 there was a small but significant increase in the ratio between serum IGF-I and IGFBP-3 levels compared with the preop ratio (P < 0.05 and < 0.005, respectively). WLB demonstrated almost complete absence of IGFBP-3 by day 2. This discrepancy between RIA and WLB analysis of IGFBP-3 suggested the presence of IGFBP-3 protease activity between days 1 and 4. This was confirmed by WLB and immunoblot analyses of samples taken 2 days after surgery. The decrease in IGFBP-3 on WLB was shown to be associated with an increase in the proteolytically cleaved fragments of IGFBP-3. These fragments following FPLC were detected in the high molecular weight fractions, suggesting that the fragments were still able to form the high molecular weight IGFBP-3/ALS complex which is thought to form only when IGF is bound by IGFBP-3. IGFBP-1 levels rose during surgery (mean duration of surgery was 125 minute) from 18 +/- 3 (preop) to 51 +/- 12 micrograms/l (end surg) (P < 0.05). This rise in IGFBP-1 paralleled increases in insulin from 7.3 +/- 1.0 to 20.8 +/- 7.5 mU/l and glucose from 4.6 +/- 0.3 to 8.7 +/- 1.2 mmol/l. IGFBP-1 levels then fell to basal values by 6 hours. IGFBP-2, in contrast, fell slightly during surgery from 636 +/- 14 to 599 +/- 96 mg/l and then returned to basal levels by 6 hours. CONCLUSION: After major surgery there are complex and diverse changes in the IGFs and IGFBPs. The effect of these changes on IGF bioavailability may significantly affect the therapeutic potential of IGF-I in this setting. PMID- 8706301 TI - Ultrasound measurement of bone. PMID- 8706302 TI - Investigation of osteoporosis. PMID- 8706303 TI - Risk factors for secondary hyperparathyroidism in a nursing home population. AB - OBJECTIVE: Secondary hyperparathyroidism may cause bone loss and structural deterioration of bone and may thus be a cause of fracture in the elderly. Vitamin D deficiency, renal impairment and medications are potential causes of hyperparathyroidism and may also directly predispose to fracture. We present the first findings of an ongoing study of hip fracture, vitamin D deficiency and hyperparathyroidism in a large Australian nursing home. DESIGN: Descriptive prevalence study. PATIENTS: Two hundred and fifty-one nursing home residents were eligible for inclusion. Informed consent and successful venepuncture were obtained for 99. Residents were of median age 83 years with interquartile range (IR) 77-89 years. MEASUREMENTS: 25-Hydroxyvitamin D (25OHD), intact parathyroid hormone (PTH), creatinine and biochemistry, demographic data and current medications. RESULTS: Fifty-two per cent of 99 subjects had 25OHD below the reference range of 28-165 nmol/l and 96.5% were below the reference range mean. Those with low 25OHD had lower plasma calcium corrected for albumin than those with normal 25OHD (medians 2.34 vs 2.41 mmol/l, 95% confidence interval for the difference between medians (CI) -0.10 to -0.04 mmol/l, P = 0.0001) and higher PTH (medians 5.8 vs 3.9 pmol/l, CI 0.10-2.6 pmol/l, P = 0.0360). Twenty-eight per cent of 97 residents had PTH above the upper reference range limit of 6.5 pmol/l. Residents receiving frusemide had higher PTH than other residents (medians 6.95 vs 3.45 pmol/l, CI 1.9-4.2 pmol/l, P < 0.0001). In linear modelling, the most important predictor of the natural logarithm of PTH was daily frusemide dose, adjusted R2 (Ra2) = 31.8%, F = 39.3, P < 0.001. Creatinine and the reciprocal of 25OHD were other significant predictors with the final Ra2 = 39.4%, F = 17.7, P < 0.001. CONCLUSIONS: Vitamin D deficiency is a common risk factor for secondary hyperparathyroidism in nursing home residents despite a climate in which vitamin D nutrition is thought to be ample. However, the daily frusemide dose is a more important predictor of PTH in this population. PMID- 8706304 TI - Bone alkaline phosphatase and collagen markers as early predictors of height velocity response to growth-promoting treatments in short normal children. AB - OBJECTIVE: A number of long-term research studies are in progress to evaluate the effects of treatment with GH on growth and final height in children with short stature but no demonstrable abnormality of GH secretion. Such treatment is invasive, expensive and carries some risk to the child. An early indication of growth response would allow restriction of treatment to those children most likely to benefit, but anthropometric measurements are relatively subjective, insensitive and imprecise. The aim of this study was to evaluate bone alkaline phosphatase, procollagen Type I C-terminal propeptide, procollagen Type III N terminal propeptide and the cross-linked carboxy-terminal telopeptide of Type I collagen as early biochemical predictors of height velocity response to growth promoting treatments in short normal children. DESIGN: A prospective intervention study, partially placebo controlled on a double blind basis. PATIENTS: Fifty healthy children with familial short stature or constitutional delay in growth and puberty (8 girls, 42 boys, ages 5.5-16.5 years and all either prepubertal (45) or in very early puberty (5 boys) at the start of treatment) were treated with placebo (6), GH alone (32), GH plus oxandrolone (8) or GH plus testosterone (4). MEASUREMENTS: Bone alkaline phosphatase and the collagen markers were measured at the start of treatment and 3 months later. Height velocity was calculated at the start of treatment and again after one year. RESULTS: Pre treatment biochemical marker concentrations did not predict height velocity response after one year. Increments in all markers after 3 months were significantly correlated with height velocity increments after one year of treatment, the highest correlations being observed for bone alkaline phosphatase (r = 0.67, P < 0.0001) and procollagen Type III N-terminal propeptide (r = 0.57, P < 0.0001). Highly significant correlations (P < 0.0001) were also observed between bone alkaline phosphatase and procollagen Type I C-terminal propeptide (r = 0.55) and between procollagen Type III N-terminal propeptide and the cross linked carboxy-terminal telopeptide of Type I collagen (r = 0.62). Multiple linear regression with stepwise selection of variables identified bone alkaline phosphatase and procollagen Type III N-terminal propeptide as the only two independent variables that contributed significantly to the prediction of height velocity response after one year (analysis of variance, P < 0.0001). Together they predicted 59% of the variability in height velocity response after a year. CONCLUSIONS: The best early predictors of height velocity response were bone alkaline phosphatase (a protein found in hypertrophic chondrocytes in the epiphyseal growth plate, in calcifying matrix vesicles and in mature osteoblasts) and procollagen Type III N-terminal propeptide, a marker of interstitial fibril biosynthesis in soft tissues. Using these markers, GH treatment could be targeted to those children most likely to benefit in the medium term. PMID- 8706305 TI - Comparison of methods to estimate body fat in growth hormone deficient adults. AB - OBJECTIVE: All of the presently used methods for in-vivo determination of body composition have inherent methodological errors and depend on various assumptions. We have therefore compared several different methods used to measure body fat in adult GH deficiency during GH treatment. DESIGN: Comparison of body composition data from a two-phase trial with an initial placebo-controlled, double-blind 6-month period, followed by open treatment with GH until all patients had received GH for 12 months. PATIENTS: Twenty-five patients with known GH deficiency entered the study. Baseline examinations were complete in 23 patients, and 22 patients (16 males, 6 females) completed all examinations after treatment. MEASUREMENTS: Body fat calculated from total body potassium (TBK) by whole-body 40K counting, total body water (TBW) by tritium dilution, total body nitrogen (TBN) by neutron activation, and bioelectric impedance (BIA) measurements were compared to body fat determinations by dual-energy X-ray absorptiometry (DEXA) in two-compartment and multicompartment body composition models. RESULTS: At baseline, DEXA fat mass agreed well at group level with measurements based on TBW or TBK alone, in a four-compartment model based on TBK and TBW, and a multicompartment model based on bone mineral (by DEXA), TBN and TBW. Body fat by BIA agreed less well. After 12 months of GH treatment, body fat decreased by all methods used. This decrease was smaller by DEXA than by the other methods. The four-compartment model based on TBK and TBW, and TBW alone, showed the best agreement with changes in DEXA fat. CONCLUSION: All methods showed a decrease of body fat with GH treatment, but variation between methods was considerable. PMID- 8706306 TI - The development, reliability and validity of a disease specific quality of life model for adults with growth hormone deficiency. AB - OBJECTIVE: Adults with GH deficiency frequently complain of lack of energy, fatigue, social isolation and problems with sexual relationships resulting in a low perceived quality of life. Previous studies of quality of life (QOL) in GH deficient adults have involved small numbers of patients and used measures not specifically designed for this patient population. We have devised a health related QOL model specifically designed for use in adults with GH deficiency and to assess the impact of future GH replacement therapy. DESIGN: Six measurements were chosen for inclusion in the model. Two were adapted for use after clinical interviews with 12 adult GH deficient patients: the Impact and the Life Fulfilment scales. The others were the Nottingham Health Profile, the Hospital Anxiety and Depression Scale, the Self-Esteem Scale and the Mental Fatigue Questionnaire. The reliability of the 6 measures was assessed by 2 methods: test re-test correlation and internal consistency (Cronbach's alpha). The validity of the Impact and Life Fulfilment scales was assessed by correlation with the other 4 scales. PATIENTS: Questionnaires were completed by 32 adults with hypothalamic pituitary disorders and GH deficiency (11 male, mean age 35.1 years), with a stimulated maximum serum GH response less than 10 mU/l (mean 2.96). Two had previously received GH injections in childhood. The questionnaires were also completed by 32 age and sex matched control subjects. RESULTS: The 6 scales had test re-test correlations of 0.70-0.92 indicating reliability over time. The Impact and Life Fulfilment Scales and the Mental Fatigue Questionnaire had Cronbach's alpha scores of greater than 0.6 indicating their potential for use in clinical trials. The Impact and Life Fulfilment scales correlated significantly with many physical and psychological domains from the other 4 scales indicating these were valid in the assessment of health related QOL in GH deficient adults. Compared to the controls the patients with GH deficiency were significantly psychosocially disadvantaged in terms of depression, self-esteem, mental fatigue and life fulfilment. CONCLUSION: The results of the reliability and validity studies indicate that this health related quality of life model for use with adults with GH deficiency is a potentially valid and reliable tool that could be used to assess the effect of GH treatment. PMID- 8706307 TI - Characterization of inhibin immunoreactivity in post-menopausal women with ovarian tumours. AB - BACKGROUND AND OBJECTIVE: We have previously reported elevated serum immunoreactive inhibin (INH) levels in patients with ovarian malignancies, particularly granulosa cell and mucinous tumours. The present study was designed to compare INH measurements using a heterologous radioimmunoassay with cross reactivity for inhibin alpha-subunit derived peptides with measurements obtained using a new ELISA specific for dimeric inhibin-A. It was hypothesized that granulosa cell tumours may secrete significant quantities of inhibin-A whereas mucinous tumours were unlikely to do so because of the lack of a relation between INH and FSH measurements in the latter group. DESIGN: Serum samples obtained from women found to have ovarian cancer were assayed using the heterologous radioimmunoassay (the Monash assay) and using an ELISA specific for dimeric inhibin (the Groome assay) and the results were compared. PATIENTS: Samples for assay were available from 69 normal post-menopausal control women, 12 patients with mucinous tumours of the ovary, 26 with serous tumours, 7 with granulosa cell tumours and 8 with various other ovarian tumours. Patients were post-menopausal or had been subjected to bilateral oophorectomy at the time these samples were collected. MEASUREMENTS: The Monash and Groome assays were carried out as described previously. The upper limit of normal for post-menopausal women in the Monash assay was 122 U/l and for the Groome assay was calculated to be 32 ng/l. RESULTS: Among the 69 normal subjects, 4 were found to have elevated inhibin levels using the Monash RIA (133-190 U/l) and 4 were found to have elevated levels in the Groome ELISA (45.5-55.3 ng/l). Among 12 patients with mucinous tumours, 10 (83%) had elevated inhibin levels using the Monash assay but only 3 (25%) had elevated levels with the Groome assay (P < 0.005). Among 26 with serous tumours, 15 (58%) had elevated levels in the Monash assay but only 1 (4%) in the Groome assay (P < 0.001). Among 7 samples from patients with granulosa cell tumours, 100% were elevated in the Monash assay and 71% in the Groome assay (NS, non-significant). In a miscellaneous group of tumours all 8 had elevated levels in the Monash assay and 2 in the Groome assay (P < 0.001). CONCLUSIONS: It was concluded that in normal postmenopausal subjects, INH is generally undetectable or present at low levels, consistent with the loss of ovarian function. The majority of granulosa cell tumours appear to secrete significant amounts of dimeric inhibin-A, whereas mucinous tumours secrete predominantly other forms of INH, presumably related to the alpha-subunit. Serous tumours may also secrete inhibin-related peptides but not dimeric inhibin-A. The nature of the inhibin related peptides produced by epithelial ovarian cancers remains to be characterized. PMID- 8706308 TI - The maternal hypothalamic-pituitary-adrenal axis in the third trimester of human pregnancy. AB - OBJECTIVE: The third trimester of pregnancy is characterized by a mildly hyperactive hypothalamic-pituitary-adrenal (HPA) axis, possibly driven by elevated circulating levels of corticotrophin releasing hormone (CRH) of placental origin. In-vitro studies have demonstrated that glucocorticoids and oestrogen stimulate while progesterone inhibits the expression of CRH mRNA and/or protein, suggesting that several potential interactions between the placenta and the HPA axis may exist. DESIGN AND PATIENTS: To investigate the detailed pattern of circulating immunoreactive (ir) CRH, ACTH, cortisol, oestradiol and progesterone during the third trimester of pregnancy, plasma samples were drawn serially every 30 minutes from 22 healthy pregnant women (age 32.0 +/- 1.1 years, mean +/- SE) between the 34th and 36th week of gestation. Ten women had plasma samples drawn between 0800 h and 2000 h (daytime group), and 12 between 2000 h and 0800 h (night-time group). The hormone concentrations obtained were analysed for pulsatility by the Detect program, for detection of circadian rhythmicity by comparison between the first and second 6-hour periods within each group by Student's t-test, and for time-dependent correlations by cross-correlation analysis. RESULTS: All five hormones were secreted in a pulsatile fashion. There was no apparent circadian rhythm of CRH or oestradiol secretion, whereas there was a clear circadian rhythm in plasma ACTH, cortisol and progesterone secretion, with the latter in reverse phase (P < 0.05). No significant correlations were observed between CRH and ACTH, whereas, as expected, ACTH and cortisol concentrations were strongly correlated with each other over time (r = 0.32 and 0.70 at lag time 30 minutes for the daytime and night-time groups, respectively), with ACTH leading cortisol. A weak positive correlation was observed between CRH and cortisol concentrations for the night-time group at lag time 0 minute, suggesting that the latter may have a positive effect on the former in vivo. CONCLUSIONS: These data suggest that placental CRH, although pulsatile, drives quantitatively the maternal HPA axis in the third trimester of pregnancy in a non circadian, non-pulsatile fashion. The maternal HPA axis is probably driven in a circadian and pulsatile fashion by another major ACTH secretagogue, most likely AVP of parvocellular paraventricular nucleus origin. PMID- 8706309 TI - Determination of renin, angiotensin converting enzyme and angiotensin II levels in human placenta, chorion and amnion from women with pregnancy induced hypertension. AB - OBJECTIVE: Pregnancy induced hypertension has been shown to be associated with a normal or low activity of the maternal circulating renin-angiotensin system (RAS) but little is known of the local RAS in placenta and fetal membranes. The present study attempts to determine, at full term of human preeclamptic pregnancies, the activity of the chorioplacental renin-angiotensin system. PATIENTS AND MEASUREMENTS: We analysed the concentrations of active renin, prorenin, angiotensin converting enzyme (ACE) and angiotensin II in homogenates of human placenta and fetal membranes from preeclamptic patients at full term pregnancy. The values of renin, ACE and angiotensin II found in the patients with moderate preeclampsia (gestosis index 0-1) (n = 10) were compared with those of normal pregnant women (n = 8). RESULTS: Our experiments showed that in preeclamptic pregnancies, the chorion membrane contained the highest concentrations of active renin (2905 +/- 152 pg/g, mean +/- SD), prorenin (21,315 +/- 2849 pg/g) and ACE (1258 +/- 302 U/g) whereas the placenta had more angiotensin II than the chorion and amnion (741 +/- 45 vs 456 +/- 40 and 428 +/- 64 pg/g, respectively). In the placenta, as in the fetal membranes, no significant difference was found in the levels of active renin, ACE or angiotensin II between hypertensive patients and normal subjects but a slightly lower level of chorionic prorenin (P < 0.05) was observed in pregnancy induced hypertension. CONCLUSION: These findings indicate that in moderate preeclampsia (gestosis index 0-1), the activity of the renin angiotensin system in term human placenta and fetal membranes remains essentially normal. PMID- 8706310 TI - Steroids in human intrauterine fluids of early pregnancy. AB - OBJECTIVE: Little is known of the hormone environment of the developing early human embryo. We have therefore measured selected steroids in the intrauterine fluids of early pregnancy. DESIGN: Measurement of progesterone, 17 alpha hydroxyprogesterone, oestradiol-17 beta, testosterone, androstenedione, cortisol and dehydroepiandrosterone sulphate in matched samples of coelomic fluid, amniotic fluid and maternal serum collected before pregnancy termination from 12 women between 8 and 12 weeks gestation. RESULTS: Mean concentrations of progesterone, oestradiol-17 beta and 17 alpha-hydroxyprogesterone in coelomic fluid were respectively 20, 6 and 2 times greater than in maternal serum and 8, 13 and 2.6 times those in amniotic fluid. Concentrations of testosterone and androstenedione were highest in maternal serum and lowest in amniotic fluid. Cortisol and dehydroepiandrosterone sulphate were found in intrauterine fluids only at the limit of detection but in normal concentrations in maternal serum. CONCLUSIONS: Coelomic fluid contains relatively high concentrations of progesterone, oestradiol-17 beta and 17 alpha-hydroxyprogesterone which may be synthesized locally. Amniotic fluid contains lower concentrations of steroids (other than progesterone) than are found in coelomic fluid or maternal serum. Free diffusion of steroids across the amnion appears limited. This may constitute a mechanism to protect the embryo from unwanted exposure to biologically active steroids. PMID- 8706311 TI - Thyroid peroxidase: evidence for disease gene exclusion in Pendred's syndrome. AB - OBJECTIVE: Pendred's syndrome is an association between congenital neurosensory deafness and goitre with abnormal discharge of iodide following perchlorate challenge, indicating a defect of iodide organification. Although Pendred's syndrome may cause up to 7.5% of all cases of congenital deafness, the molecular basis of the association between the hearing loss and the thyroid organification defect remains unknown. We chose to investigate the role of the thyroid peroxidase (TPO) gene as the genetic defect in Pendred's syndrome. DESIGN: A highly informative variable number tandem repeat (VNTR), located 1.5 kb downstream of exon 10 of the TPO gene, was used to search for genetic linkage in multiple sibships affected by Pendred's syndrome. PATIENTS: Seven kindreds were recruited from the UK, each with at least two affected members. We have also examined a large inbred Israeli family with two affected offspring and five unaffected children. MEASUREMENTS: Individuals were assigned affected status based on the characteristic clinical features of Pendred's syndrome, namely the presence of congenital sensorineural hearing loss and the appearance in early life of a goitre. Additionally, at least one affected member from each sibship had a characteristic positive perchlorate discharge test (Morgans & Trotter, 1958). PCR amplification of genomic DNA at the TPO VNTR allowed assignment of genotypes to each individual and the calculation of a two-point LOD score. RESULTS: In six of the nine sibships analysed we found obligatory recombination between TPO and Pendred's syndrome. Non-complementation observed in affected parents with an affected offspring excluded TPO in an affected sibship with genotype sharing and supports a hypothesis of genetic homogeneity for Pendred's syndrome. In two sibships, mutation of the TPO gene as the cause of Pendred's syndrome could not be excluded. CONCLUSIONS: These data suggest that defects at the thyroid peroxidase locus on chromosome 2 are not the major cause of Pendred's syndrome. PMID- 8706312 TI - Urinary excretion of pyridinoline and deoxypyridinoline measured by immunoassay in hypothyroidism. AB - OBJECTIVE: We measured pyridinium cross-links, markers of bone resorption, by an enzyme-linked immunosorbent assay (ELISA) in hypothyroid patients to see whether bone resorption was reduced in hypothyroidism and whether it increased with T4 treatment. DESIGN AND PATIENTS: Eight hypothyroid patients, whose initial TSH levels were 268.1 +/- 87.7 mU/l (mean +/- SE), were treated with T4 (100 micrograms/day). Urinary excretion of pyridinium cross-links was assayed before and after T4 treatment. MEASUREMENTS: Pyrilinks and Pyrilinks-D kits were used. The Pyrilinks assay measures free forms of pyridinoline and deoxypyridinoline together (PYD), while the Pyrilinks-D assay measures deoxypyridinoline (DPD) alone. The Pyrilinks reference ranges for normal subjects are 8-24nmol/mmol creatinine in males and 10-28nmol/mmol creatinine in normal premenopausal females. The DPD reference ranges obtained from normal men and women aged 40-50 years were 3.20 +/- 0.75 (mean +/- SD) nmol/mmol creatinine and 4.55 +/- 1.22 nmol/mmol creatinine, respectively. RESULTS: The sensitivity of the assay was enhanced by simply using less diluted urine samples. Concentrations of both compounds of the urinary pyridinium cross-links were low in untreated hypothyroid patients and increased gradually as thyroid hormone status improved from hypothyroidism to euthyroidism. One month after treatment when the TSH levels in the patients were still as high as 74.4 +/- 44.5 mU/l, urinary PYD excretion has increased to 2.6 times the pretreatment level. When the TSH levels of the patients decreased below 10 mU/l, both PYD and DPD increased significantly to 3.8 and 3.3 times pretreatment values, respectively. CONCLUSIONS: Although hyperthyroidism or excess treatment with thyroid hormone has been known to induce bone resorption, this is the first report that urinary excretion of pyridinium cross-links is reduced in hypothyroidism and is normalized by physiological thyroid hormone replacement. PMID- 8706313 TI - Glucose turnover, fuel oxidation and forearm substrate exchange in patients with thyrotoxicosis before and after medical treatment. AB - OBJECTIVE: Accelerated metabolism is a hallmark of thyrotoxicosis, but the underlying biochemical mechanisms are incompletely understood and the majority of studies have investigated normal subjects rendered only modestly hyperthyroid for a brief period of time. We have therefore studied a group of thyrotoxic patients using several different techniques. DESIGN: Twelve patients with newly diagnosed diffuse (10 patients) or nodular (2 patients) toxic goitre (10 women, 2 men; age 42.8 +/- 3.2 years; BMI 21.6 +/- 0.7 kg/m2) before ('pretreatment') and after ('treated') 11.2 +/- 1.0 weeks treatment with methimazole and compared these patients to a control group ('control') of 11 subjects (9 women, 2 men; age 40.5 +/- 3.9 years; BMI 22.5 +/- 1.0 kg/m2). All were studied for 3 hours in the basal state, using indirect calorimetry, isotope dilution for the measurement of glucose turnover and the forearm technique for assessment of muscle metabolism. RESULTS: Prior to treatment patients with thyrotoxicosis were characterized by increased (P < 0.05) levels of T3 (3.75 +/- 0.23 nmol/l (pretreatment), 1.89 +/- 0.08 (treated) and 1.75 +/- 0.11 (control)), resting energy expenditure (130.5 +/ 3.5 (pretreatment), 107.7 +/- 2.7 (treated) and 106.3 +/- 3.1 (control), % of predicted), protein oxidation (0.67 +/- 0.03 (pretreatment), 0.54 +/- 0.06 (treated) and 0.46 +/- 0.05 (control), mg/kg/min), lipid oxidation (1.34 +/- 0.08 (pretreatment), 1.00 +/- 0.06 (treated) and 1.02 +/- 0.04 (control), mg/kg/min), endogenous glucose production (2.51 +/- 0.13 (pretreatment), 1.86 +/- 0.12 (treated) and 1.85 +/- 0.12 (control), mg/kg/min), non-oxidative glucose turnover (1.28 +/- 0.16 (pretreatment), 0.75 +/- 0.18 (treated) and 0.71 +/- 0.11 (control), mg/kg/min) and a 50% increase in total forearm blood flow. Glucose oxidation (1.23 +/- 0.09 (pretreatment), 1.13 +/- 0.10 (treated) and 1.21 +/- 0.11 (control) mg/kg/min), exchange of substrates in the muscles of the forearm and circulating levels of insulin, C-peptide, growth hormone or glucagon were not influenced by hyperthyroidism. Propranolol (20 mg thrice daily) given to 7 of the patients for 2 days did not affect circulating levels of thyroid hormones, energy expenditure or glucose turnover rates. CONCLUSIONS: These results suggest that all major fuel sources contribute to the hypermetabolism of thyrotoxicosis and that augmented non-oxidative glucose metabolism may further aggravate the condition. All abnormalities diminish with medical treatment of the disease. PMID- 8706314 TI - The impact of liver transplantation on endocrine status in men. AB - OBJECTIVE: There are few longitudinal data on the endocrine changes which occur after liver transplantation. We have therefore studied the impact of orthotopic liver transplantation (oLTX) on the hypothalamic-pituitary-gonadal hormone axis and sex steroid metabolism in men. PATIENTS AND STUDY DESIGN: Ten male patients with end-stage liver failure due to alcohol induced cirrhosis (n = 2), virus induced cirrhosis (n = 5), primary biliary cirrhosis (n = 1) and idiopathic cirrhosis (n = 2) were included in a prospective study analysing the impact of oLTX on endocrine status. They were studied before and after oLTX with a mean follow-up of 11.6 months (range 4-23) following transplantation. MEASUREMENTS: Serum levels of LH, FSH, testosterone (TE), free TE, PRL, cortisol, oestradiol (E2) and sex hormone binding globulin (SHBG) were analysed with commercially available radioimmunoassays in all individuals before and after oLTX. Gonadotrophin releasing hormone stimulation tests were done in 5 patients before and after oLTX. Additionally, a complete urological assessment with a detailed questionnaire on sexual function was obtained from all individuals. RESULTS: Prior to oLTX, endocrine status was invariably abnormal, the most prominent finding being a pathological decrease of TE in 90% and of free TE in all cases. After successful oLTX, all individuals had physiological levels of TE and of free TE which increased twofold (P < 0.01) and tenfold (P < 0.0001), respectively. Additionally, serum gonadotrophin (LH/FSH) levels increased in the majority of patients, while E2 decreased following oLTX. Endocrine changes extended beyond the hypothalamic-pituitary-gonadal hormone axis, as shown by a decrease in PRL (P < 0.02) and SHBG (P < 0.01) after transplantation. GnRH tests revealed normal stimulation of LH and FSH before and after oLTX in all cases. Libido, potency and frequency of sexual intercourse improved significantly after oLTX in the majority of patients. CONCLUSIONS: These findings demonstrate the ability of the hypothalamic-pituitary-gonadal hormone axis and sex steroid metabolism to resume physiological function following orthotopic liver transplantation in men. Correspondingly, sexual function returns to normal in the majority of patients, despite significant alterations prior to orthotopic liver transplantation. PMID- 8706315 TI - Expression of the transferrin receptor in human anterior pituitary adenomas is confined to gonadotrophinomas. AB - OBJECTIVE: The human pituitary gland is affected selectively by conditions associated with iron deposition, but the mechanisms for this are unknown. In this study we have determined whether the transferrin receptor, which mediates iron uptake by cells, could be detected immunocytochemically in human pituitary adenomas in vitro. PATIENTS: Data were derived from 35 patients undergoing transsphenoidal surgery and included 13 with clinically non-functioning adenomas, 15 with acromegaly, 4 prolactinomas, 2 patients with Cushing's disease and one patient with Nelson's syndrome. MEASUREMENTS: Transferrin receptor immunopositivity was determined for each adenoma in dispersed cell culture using a specific monoclonal antibody. RESULTS: Eight of 13 clinically functionless adenomas showed immunopositive transferrin receptor expression, whilst adenomas from 15 patients with acromegaly, 4 prolactinomas, 2 Cushing's syndrome and one patient with Nelson's syndrome were negative. The eight transferrin receptor positive tumours were gonadotrophinomas and accounted for eight of the nine tumours which secreted and immunostained for FSH; all eight also secreted and immunostained for LH. CONCLUSIONS: These findings may reflect a special requirement for iron by gonadotrophin secreting cells in comparison to other pituitary cell types and this could underlie the reasons why in the normal pituitary these cells are especially susceptible to malfunction in iron overload syndromes such as genetic haemochromatosis and beta-thalassaemia. PMID- 8706316 TI - A comparison between short ACTH and insulin stress tests for assessing hypothalamo-pituitary-adrenal function. AB - OBJECTIVE: Insulin-induced hypoglycaemia is the standard method for assessment of the hypothalamo-pituitary-adrenal (HPA) axis of patients with pituitary or hypothalamic disease. It has been claimed that a normal cortisol response to the 30-minute ACTH stimulation test (AST) obviates the need to perform the insulin stress test (IST) in these patients. The objective of our study was to compare both tests in a group of consecutive patients with pituitary disease. SUBJECTS AND METHODS: Thirty patients with pituitary disease were evaluated by standard IST (0.1 U of soluble insulin/kg body weight, i.v.) after fasting from midnight and AST (250 micrograms synacthen, i.v.). In the IST, a plasma glucose of < 2.2 mmol/l was taken as the hypoglycaemic threshold and blood was collected at 0, 30, 60, 90 and 120 minutes. In the AST blood was collected at 0 and 30 minutes. Serum cortisol was measured by standard radioimmunoassay and glucose by the glucose oxidase method. Cortisol responses to the stimuli were compared at cut-off levels of 550, 500, 450 and 400 nmol/l. RESULTS: At 550 nmol/l, out of 30 patients, 17 showed an abnormal IST of whom 9 had normal responses to AST (53%). At 500 nmol/l, 12 patients had an abnormal IST of whom 6 had normal AST (50%). At 450 nmol/l, of 9 patients with an abnormal IST, 5 had a normal AST (56%). At 400 nmol/l, 5 patients had an abnormal IST all of whom (100%) showed a normal AST. CONCLUSION: There is a clear discrepancy between the results of the two tests at different cortisol cut-off levels. The ACTH stimulation test is not reliable for assessing the HPA axis in patients with pituitary disease and the insulin stress test remains the standard method. PMID- 8706317 TI - Steroid 5 alpha-reductase 2 deficiency: virilization in early infancy may be due to partial function of mutant enzyme. AB - Male pseudohermaphroditism due to steroid 5 alpha-reductase deficiency is the consequence of mutations in the gene encoding the type 2 isoenzyme. Most (60%) affected subjects have homozygous mutations, and the remainder are compound heterozygotes or presumed compound heterozygotes. We report an Italian subject with phenotypic and endocrine features of 5 alpha-reductase 2 deficiency who is homozygous for a substitution mutation (H231R). Although close consanguinity is not present, genealogical data demonstrated that the parents are distantly related, and both parents and the maternal grandmother are heterozygous carriers of the mutation. The fact that this particular mutation results in the formation of an enzyme with considerable residual activity may explain in part the significant degree of virilization that took place in this subject in early infancy. This same mutation (H231R) is present in heterozygous form in two other families, an African-American family and an American family of northern European descent. PMID- 8706318 TI - ACTH independent Cushing's syndrome occurring in siblings. AB - Familial Cushing's syndrome due to ACTH independent bilateral macronodular adrenocortical hyperplasia occurring in siblings is reported. The proband was a 69-year-old woman who presented with a typical Cushingoid appearance. The serum cortisol level was elevated, with a loss of diurnal rhythm, and the plasma ACTH level was undetectable. Dynamic testing showed no suppression of urinary 17-OHCS by high dose dexamethasone and no stimulation by metyrapone. An abdominal CT scan showed bilateral adrenal enlargement. The patient died of a subarachnoid haemorrhage, and autopsy revealed a massively thickened adrenal cortex composed of nodules up to 3.5 cm in diameter. A pituitary adenoma was not found. We learned that the patient's elder brother was also diagnosed at 59 years of age with Cushing's syndrome due to bilateral macronodular adrenocortical hyperplasia. His plasma cortisol levels were not suppressed by high dose dexamethasone and the plasma ACTH level was undetectable. Screening of the available family members by administering 1 mg dexamethasone at midnight and performing abdominal CT scan revealed impaired suppressibility of serum cortisol associated with enlarged bilateral adrenal glands in a 64-year-old sister and a 54-year-old brother. The 64-year-old sister was considered as a possible 'affected' case in the early stages of development, because the basal level of ACTH was not suppressed and hyperplasia of the bilateral adrenal glands as revealed by CT scan was less evident. PMID- 8706319 TI - Short stature, GH deficiency and social status. PMID- 8706320 TI - Effects of gonadal androgens on adrenal androgen levels. PMID- 8706321 TI - Immunological control in primary biliary cirrhosis: the cytokine story. PMID- 8706322 TI - Helicobacter pylori infections in IgA deficiency: lack of role for the secretory immune system. AB - During the last years considerable attention has been focused on the possibility of the development of an oral vaccine against Helicobacter pylori. However, Helicobacter infection is known to be life-long, despite a vigorous immune response, and the hypothesis that an increased local production of secretory IgA in the gastric mucosa, due to vaccination, should protect against the colonization of the bacterium may therefore be questioned. In this study, when comparing the seropositivity and titre against H. pylori in IgA-deficient patients and age-related normal blood donors, it appears that lack of secretory IgA does not seem to have any major influence on the prevalence of the infection, nor is it reflected in titres of specific IgG antibodies. These results many argue against a pivotal role for IgA in the defence against Helicobacter, and raise questions about current strategies for the development of an oral vaccine against H. pylori and may point to a need for alternative therapeutic strategies. PMID- 8706323 TI - Chronic mucocutaneous candidiasis. I. Altered antigen-stimulated IL-2, IL-4, IL-6 and interferon-gamma (IFN-gamma) production. AB - Patients with chronic mucocutaneous candidiasis (CMC) present with persistent infections with the opportunistic yeast Candida. Impaired cell-mediated responses to Candida have been documented in CMC patients, but the defect remains poorly understood. The importance of Th1 cytokines in resistance and Th2 in susceptibility to Candida infections has recently been demonstrated in murine models. In our studies we evaluated production of IL-2 and IFN-gamma (markers of Th1 type responses) as well as IL-4 and IL-6 (Th2 type markers) following stimulation with two kinds of Candida antigens (CAgs), polysaccharide antigens, tetanus toxoid and pokeweed mitogen. Our results demonstrate that CMC patients have impaired cytokine production upon in vitro stimulation with CAgs resulting in low or absent IL-2, increased IL-6 and either absent or increased IFN-gamma production. Cytokine production following stimulation by other antigens was unaltered. The overall cytokine-producing capacity assessed through mitogen stimulation was also intact. Addition of IFN-alpha or IFN-gamma to culture in an attempt to modify cytokine production did not have significant effects. Levels of soluble IL-6 receptors were not increased and could not account for increased IL 6 production. Our studies support the hypothesis that Candida antigens trigger a predominantly Th2 instead of a Th1 cytokine response in patients with CMC. PMID- 8706324 TI - Chronic mucocutaneous candidiasis. II. Class and subclass of specific antibody responses in vivo and in vitro. AB - Patients with chronic mucocutaneous candidiasis (CMC) succumb to persistent infections with the opportunistic yeast Candida. Impaired cell-mediated responses to Candida have been repeatedly reported while antibody responses were mostly found to be normal. The underlying defect remains poorly understood. It has recently been shown that CMC patients are also susceptible to infections with encapsulated bacteria, and may have associated IgG2 and IgG4 deficiency. Our previous studies demonstrated altered cytokine production in CMC patients. As cytokines can influence production and isotype of specific antibody, in 10 patients with CMC we measured the levels and isotype distribution of serum antibodies to Candida antigens (CAg), pneumococcal polysaccharide (PPS) and tetanus toxoid (TT) antigens. Peripheral blood lymphocytes were also stimulated in culture and the antibodies made in vitro were measured. Our data demonstrated that in vivo, CMC patients had very high levels of IgG and IgA CAg-specific antibodies. CAg-specific and PPS-specific IgG1 was markedly higher than in controls. Children but not adults with CMC had significantly lower levels of IgG2 specific antibody to CAg and PPS compared with age-matched controls. Patients had significantly higher levels of IgG3-specific antibody to all three antigens tested. These findings were in accordance with increased total IgG and IgG3 levels seen in CMC patients. In vitro, CMC patients, particularly children, did not respond as frequently to antigen stimulation as did their healthy controls. The level of specific antibody produced was also lower to all antigens tested, as was the amount of total immunoglobulins following antigenic and particularly mitogenic stimulation. Addition of interferon-alpha (IFN-alpha) or IFN-gamma to cultures had variable, sometimes marked, effects. Our results demonstrate that CMC patients manifest subtle alterations in specific antibody responses to CAg, PPS and TT, which are most pronounced in children. This may relate to altered cytokine production also seen in these patients. PMID- 8706325 TI - CD8+ lymphocyte phenotype and cytokine production in long-term non-progressor and in progressor patients with HIV-1 infection. AB - In most HIV-1-infected patients, clinical and immunological progression develops within a few years. Few infected people, termed long-term non-progressors (LTNP), remain healthy and immunologically stable for a long time. The factors governing the maintenance of this condition are not well known, but it is conceivable that CD8+ lymphocytes, cells that play a central role in controlling in vitro HIV replication, may have a part in vivo in this process. The aim of this study was to characterize the phenotypic profile and the cytokine production of CD8+ cells in a group of LTNP patients who had stable CD4+ cell counts (> 500/mm3) for at least 7 years. Their CD8+ absolute numbers were similar to a control group composed of HIV-1+ patients who have a progressive decline of their CD4+ cell counts. However, our multiparameter immunofluorescence studies show that a clinical and immunologically stable condition is associated with the presence of a CD28+, CD95 strongly positive CD8+ population, while disease progression is marked by the CD28-CD95+CD8+ subset. Purified CD8+ cells from LTNP retain their ability to produce IL-2, interferon-gamma (IFN-gamma) and, to a lesser degree, to produce IL-10 and IL-4. In contrast, CD8+ cells from progressors are unable to secrete IL-2 and IL-10. Although CD8+ cytokine profile does not fit with the proposed T helper (Th)1/Th2 switch in progressive HIV infection, LTNP CD8+ T cells maintain their capacity to produce IL-2 and IL-10 (Th0-like), a pattern very similar to that observed in normal HIV healthy controls. We suggest that CD8+ cells expressing CD28, CD95 and having a Th0-like profile may be considered to be associated with long-term survival. PMID- 8706326 TI - Reversal of the inhibitory effects of HIV-gp120 on CD4+ T cells by stimulation through the CD28 pathway. AB - The effects of exposure to HIV-gp120 on proliferation and cytokine production by T cell lines were investigated. T cell lines were generated by stimulation of peripheral blood mononuclear cells from several healthy donors with cross-linked anti-CD3 antibodies and IL-2. These T cell lines exhibited the characteristics of Th1 cells, producing IL-2 and interferon-gamma (IFN-gamma), but not IL-4, on stimulation with anti-CD3 antibodies. In the presence of gp120, stimulation with anti-CD3 antibodies was inhibited in terms of both proliferative responses and the secretion of IL-2 and IFN-gamma. Similar effects were observed when a T cell line was stimulated in the presence of a synthetic peptide representing the CD4 binding region of gp120. Neither gp120 nor the CD4-binding region peptide had any effect on IL-4 production by the T cell lines. Stimulation through the CD28 pathway partially restored both the proliferative effect and cytokine production by T cell lines in response to anti-CD3 antibodies in the presence of gp120. Anti CD28 antibodies also partially restored cytokine production when purified CD4+ T cells from a T cell line were stimulated with anti-CD3 antibodies in the presence of gp120. Anti-gp120 antibodies partially or completely reversed the inhibitory effects of gp120 on T cell proliferation. These results indicate that stimulation through the CD28 pathway may restore defective CD4+ T cell responses in HIV infected individuals. PMID- 8706327 TI - Yersinia-hsp60-reactive T cells are efficiently stimulated by peptides of 12 and 13 amino acid residues in a MHC class II (I-Ab)-restricted manner. AB - Heat shock proteins (hsp) are immunodominant antigens in microbial infections. Previous work from this laboratory demonstrated that Yersinia-hsp60 (Y-hsp60) reactive CD4+ alpha beta T cells play an important role for resolution of Y. enterocolitica infections in mice. In the present study we identified two epitopes of Y-hsp60 recognized by CD4+ Th1 cell clones. The epitopes comprise 12 (214-225) and 13 (74-86) amino acid (aa) residues of Y-hsp60, and are the first described for MHC class II (I-Ab) molecules. Both epitopes are also recognized by T cells isolated from mesenteric lymph nodes from mice orogastrically infected with yersiniae. Stimulation of T cells with peptides of 12 and 13 aa residues of Y-hsp60 caused highly efficient proliferation compared with longer peptides, full length recombinant Y-hsp60, or heat-killed Yersinia (HKY). Incubation of antigen presenting cells with chloroquine blocked both peptide and HKY-triggered T cell proliferation, whereas cytochalasin B only blocked HKY-induced proliferation and to a lesser extent peptide-induced proliferation. The identified epitopes reside in a region of Y-hsp60 that is conserved between Enterobacteriaceae but highly variable when compared with murine or human hsp60. Although both epitopes are identical to the related sequence of hsp60 (GroEL) of Escherichia coli, only weak T cell responses were observed upon stimulation with GroEL of E. coli, suggesting that other factors, e.g. flanking amino acid residues, might be important for antigen processing and T cell stimulation in a class II-restricted manner. Furthermore, these observations might be of significance for the rational design of subunit vaccines. PMID- 8706328 TI - Pneumococcal antibody responses in elite swimmers. AB - The ability of elite swimmers to mount an antibody response to the pneumococcal vaccine, Pneumovax 23, was assessed at the end of an intensive 12-week training programme. Antibody titres to six pneumococcal polysaccharide types were measured in 20 elite swimmers (10 male, 10 female) aged 17-23 years and 19 sedentary age- and sex-matched students (eight male, 11 female) aged 18-23 years. Blood samples were tested 14 days apart to assess the magnitude of the antibody response and changes in serum immunoglobulin isotypes and IgG subclasses. There were no significant differences in any of the pneumococcal antibody responses to the Pneumovax between swimmers and controls, and no gender effect, either before or after vaccination. The clinically adequate response to the vaccine was greatest for the pneumococcal serotype 4, which was 97% for the total study population. There were no significant correlations between the magnitude of any of the pneumococcal antibody responses and (i) changes in the scores for the swimmers' international performance; (ii) infection rates in either swimmers or controls; (iii) any psychological variables, assessed by the Profile of Mood States (POMS) questionnaire for either swimmers or controls. Swimmers had significantly lower concentrations of serum IgG2 (P = 0.04) and IgG3 (P = 0.002) before pneumococcal vaccination. The swimmers had an increase in all immunoglobulin isotypes and IgG subclasses post-vaccination, suggesting a polyclonal response to the vaccine that was not observed in control subjects. The magnitude of the subclass responses after vaccination was significantly greater in swimmers compared with controls for IgG1 (P = 0.04), IgG3 (P = 0.04) and IgG4 (P = 0.01). The data indicated that elite swimmers undertaking an intensive training programme were capable of mounting an antibody response to pneumococcal antigens equivalent to that of age- and sex-matched sedentary control subjects, despite the swimmers having lower prevaccination levels of serum immunoglobulins. PMID- 8706329 TI - Immunoregulation in onchocerciasis: predominance of Th1-type responsiveness to low molecular weight antigens of Onchocerca volvulus in exposed individuals without microfilaridermia and clinical disease. AB - Chronic and generalized onchocerciasis is associated with suppression of the parasite-specific cellular responsiveness, while exposed individuals without parasitological and clinical evidence of infection (endemic normals) display prominent cellular reactivity to Onchocerca volvulus antigens (OvAg). In order to identify those parasite antigens which may account for this differential cellular responsiveness, total adult worm-derived OvAg were fractionated by means of preparative SDS-PAGE and blot-elution into 22 antigen fractions of continuously decreasing molecular weight. Peripheral blood mononuclear cells (PBMC) from microfilariae (mf)-positive onchocerciasis patients (n = 18) proliferated weakly in response to all OvAg fractions. In contrast, in vitro reactivity of PBMC from endemic normals (n = 9) was depressed in response to OvAg of mol. wt 200-30 kD only, while antigens of mol. wt < 30 kD induced vigorous proliferation in these individuals compared with the microfilaridermic patients (P < 0.05). Highest proliferative reactivity of cells from endemic normals was observed in response to OvAg of mol. wt 15-11 kD. Furthermore, these low mol. wt antigen fractions induced substantial production of IL-2 and interferon-gamma (IFN-gamma) in PBMC from endemic normals, but not in those from onchocerciasis patients. Cells from individuals of both groups secreted similar amounts of IL-5 in response to all OvAg fractions, with highest production again being induced by low mol. wt OvAg. In contrast, PBMC from onchocerciasis patients clearly produced more IL-10 than did cells from endemic normals. This augmented IL-10 production by PBMC from mf positive individuals was not only observed after stimulation with OvAg fractions, but was measured in unstimulated control cultures as well. IFN-gamma-specific mRNA in antigen-stimulated PBMC from endemic normals appeared to be more prominent than in cells from onchocerciasis patients. However, mRNA transcripts of IL-10 and IL-13 were clearly present in patients, but were absent or inconsistently observed in endemic normals. Our results suggest that vigorous Th1 type cellular responsiveness encountered in endemic normals is restricted to low mol. wt antigens of O. volvulus, while such reactivity will not be present in mf positive individuals. Furthermore, spontaneous production of high levels of IL-10 in onchocerciasis patients is likely to suppress Th1-type immunity, and thus may favour manifestation of chronic onchocerciasis. These traits of cellular immunity may contribute to the differential outcome of O. volvulus infection, the manifestation of clinical disease, and may also regulate the build up of acquired immunity in humans. PMID- 8706330 TI - Cytokine mRNA expression in the liver of patients with primary biliary cirrhosis (PBC) and chronic hepatitis B (CHB). AB - The expression of cytokine mRNA species was determined in liver biopsies from six normal subjects, 18 patients with PBC and 14 patients with hepatitis B e antigen (HBeAg)-positive CHB using a reverse transcriptase-polymerase chain reaction (RT PCR) technique. cDNA, obtained by reverse transcription using oligo d(T) primers, was amplified by PCR using primers specific for the coding region of seven different cytokines (IL-1, IL-2, IL-4, IL-5, IL-6, interferon-gamma (IFN-gamma), tumour necrosis factor-alpha (TNF-alpha)). The abundance of some cytokines (IL-2, IL-4, IL-5 and IFN-gamma) was also estimated by semiquantitative RT-PCR, using as standards dilutions of synthetic cytokine mRNA transcripts, that could be distinguished electrophoretically from respective native cytokine mRNAs. Hepatic inflammation was assessed by a semiquantitative histologic score and by amplification of mRNA for T cell receptor (TCR)-alpha. mRNAs for IL-1 and IL-6 were detected in only one control liver. In CHB, mRNAs for IL-1, IL-2, IL-4, IL-5 and IFN-gamma were detected in 43%, 60%, 80%, 20%, and 54% of biopsies, respectively. mRNA for IFN-gamma and IL-4, but not IL-1, tended to be associated with severe inflammation. In five biopsies semiquantitative analyses revealed increased levels of mRNA for TCR-alpha and, when transcripts were detectable, high levels of mRNA for IFN-gamma and IL-4. In PBC, mRNA for IFN-gamma was detected in 60% of biopsies, but no mRNAs for IL-1, IL-2, IL-4, IL-5, or IL-6, or for TNF-alpha, were detected. Semiquantitative analyses revealed that absolute levels of mRNA for IFN-gamma tended to correlate with the severity of hepatic inflammation. The results suggest that: (i) there may be fundamental differences in the roles that cytokines play in the hepatic inflammatory processes of PBC and CHB; and (ii) while hepatic IFN-gamma mRNA expression is not specific for PBC, IFN-gamma may play a prominent role in the immunopathogenesis of PBC. PMID- 8706331 TI - Circulating levels of IL-11 and leukaemia inhibitory factor (LIF) do not significantly participate in the production of acute-phase proteins by the liver. AB - To investigate the contribution of IL-11 and LIF to acute-phase protein (APP) production, we first analysed the effects of IL-11 and LIF on production of C reactive protein (CRP), fibrinogen, and haptoglobin by human primary hepatocytes. We also measured the serum levels of IL-11, LIF, and CRP in serum from patients with inflammatory rheumatic diseases to assess the role of these cytokines in the APP response in vivo. We included patients with conditions associated with a high APP response such as rheumatoid arthritis (RA) or spondylarthropathy (SpA), and others usually associated with a weak APP response such as systemic lupus erythematosus (SLE), in order to investigate whether these cytokines could account for the differences in APP responses. Our results showed that IL-11 and LIF induced only minimal stimulation on production of APP by human primary hepatocytes compared with IL-6, known as the major inducer. Serum levels of CRP were elevated in RA and SpA, and significantly higher than in SLE patients. Despite the presence of a high APP response in some of our patients and despite the fact that we used sensitive assays to measure IL-11 and LIF, serum levels of both cytokines were not detected in any of the tested sera. In conclusion, our results show that circulating levels of IL-11 or LIF do not contribute significantly to the production of APP in vivo, and that they do not account for the difference in APP response between SLE and other inflammatory rheumatic diseases. PMID- 8706332 TI - Reduction in serum levels of antimitochondrial (M2) antibodies following immunoglobulin therapy in severe combined immunodeficient (SCID) mice reconstituted with lymphocytes from patients with primary biliary cirrhosis (PBC). AB - The effect of gammaglobulin treatment on autoantibody production was investigated in SCID mice reconstituted with human peripheral blood mononuclear cells (PBMC) obtained from patients with PBC. All reconstituted mice displayed the presence of human antimitochondrial antibodies (alpha M2Ab) of both IgG and IgM types before treatment with human immunoglobulin. Two weeks after i.p. injection of 20 x 10(6) PBMC into SCID mice, i.p. treatment with various preparations of human immunoglobulin was initiated. In control animals treated with saline, serum levels of human alpha M2Ab of the IgG type increased with time, peaking around 4 weeks after reconstitution. In contrast, human IgG autoantibodies rapidly decreased in all animals treated with human IgG. Treatment with a human IgM preparation had no effect on serum levels of alpha M2Ab of the IgG type. The results may suggest that the pronounced reduction of specific IgG autoantibodies was due to an increased catabolism of human IgG, including the autoantibodies, in the gammaglobulin-treated mice. Although the production of human alpha M2Ab in reconstituted mice could be easily shown, PBC-specific liver lesions or bile duct destruction were not observed, irrespective of treatment protocol. PMID- 8706333 TI - IL-4 enhances IEC-6 intestinal epithelial cell proliferation yet has no effect on IL-6 secretion. AB - Intestinal epithelial cells (IEC) form an important line of defence at the intestinal mucosa by providing a barrier to lumenal contents and also by their ability to secrete various inflammatory cytokines. Recently, several T cell derived cytokines have been shown to regulate specific IEC functions. In this study, the effect of IL-4 on IEC proliferation and secretion of the inflammatory cytokine IL-6 was investigated using the non-transformed rat IEC-6 intestinal epithelial cell line. Recombinant rat (rr)IL-4 was found to enhance IEC-6 cell proliferation over 4 days of culture, and this enhancement was dose-dependent. Further studies using specific antibodies confirmed that IL-4 induced the effect and that the effect was not mediated by autocrine-produced transforming growth factor-alpha. However, IL-4 did not induce IL-6 secretion by the IEC-6 cells, nor did it alter IL-1 beta-induced IL-6 secretion. These results indicate that T cells may be capable of regulating IEC proliferation via the secretion of IL-4 without altering the capacity of the IEC to function in the inflammatory response by secreting IL-6. PMID- 8706334 TI - Crohn's disease (CD) patients suffering from peripheral arthritis or ankylosing spondylitis reveal restricted T cell receptor V beta regions in different temporal phases of disease. AB - Little is known about the mechanisms triggering and controlling both the development and perpetuation of extraintestinal complications in CD. The aim of the present study was to test the hypothesis that the T cell immune response in CD patients with joint complications may be altered when compared with patients without extraintestinal manifestations. We used a semiquantitative polymerase chain reaction assay to analyse the T cell antigen receptor repertoire in peripheral blood T cells from eight CD patients suffering from peripheral arthritis and ankylosing spondylitis, 12 CD patients without extraintestinal manifestations, and from seven non-CD patients with ankylosing spondylitis showing typical changes on joint radiographs. Being concerned that different patterns may be seen in different phases of the inflammatory disease process, we have also taken care to analyse sequential samples at various time points of the disease. Expression of all 22 V beta genes was found in each healthy control and in each CD patient without extraintestinal manifestations and showed no major variation over time. Southern hybridization analysis of amplified products revealed a highly restricted V beta repertoire in all CD patients suffering from peripheral arthritis and ankylosing spondylitis. In contrast, non-CD patients with ankylosing spondylitis without signs or symptoms of gastrointestinal problems demonstrated the presence of the entire V beta repertoire. Our longitudinal studies confirmed variable V beta usage over time, as certain transcripts were found only in distinct temporal phases of disease. Our data are not directly suggestive of a common superantigen model of CD, but instead emphasize a specific decrease in signals throughout the T cell receptor V beta repertoire in CD patients suffering from joint complications. PMID- 8706335 TI - Clinical features of anti-chromo antibodies associated with anti-centromere antibodies. AB - Anti-chromo antibodies (AChA) are autoantibodies accompanying anti-centromere antibodies (ACA). We determined the frequency and clinical significance of AChA in autoimmune rheumatic diseases. Serum samples from 252 patients with rheumatic diseases were examined by immuno-blotting with HeLa nuclear extract and with recombinant N-terminus of 25-kD chromo protein (p25). AChA were detected in 28 (36%) of 77 sera with ACA. AChA were found only in ACA-positive sera. Twenty-two (79%) of 28 recognized a recombinant N-terminal portion of p25, including the chromo domain which is conserved among species. AChA were related to leucopenia, thrombocytopenia, elevated erythrocyte sedimentation rate, and existence of Sjogren's syndrome (SS). In ACA-positive patients, AChA might be a serologic indicator of systemic sclerosis (SSc), having features of systemic lupus erythematosus and/or SS or diseases other than SSc. PMID- 8706336 TI - Characterization of altered calcium signalling in T lymphocytes from patients with rheumatoid arthritis (RA). AB - Abnormal function of peripheral blood T lymphocytes is characteristic of RA; diminished proliferation and secretion of cytokines following in vitro mitogen stimulation are observed. We have investigated the calcium flux initiating T cell activation in rheumatoid peripheral blood mononuclear cells (PBMC) to determine whether abnormalities in signalling are also present. We have found that both phytohaemagglutinin (PHA-P)- and anti-CD3-stimulated calcium fluxes were much reduced in the patients' PBMC compared with controls, with a mean six-fold difference (P < 0.01) in rate of Ca2+ flux with PHA-P stimulation. When purified T cells were examined with PHA and CD3 stimulation, a reduction in the peak and plateau [Ca2+]i was observed in RA T cells, but the rate of rise of [Ca2+]i was only reduced in those cells stimulated with PHA. These results suggest that alterations in the initiating signal may underlie the functional T cell abnormalities associated with RA, and that there may be an additional extrinsic influence from non-T cells in the PBMC population. PMID- 8706337 TI - Autoantibodies to mitochondrial 2-oxo-acid dehydrogenase complexes in localized scleroderma. AB - Sera from patients with localized scleroderma frequently produce cytoplasmic staining by indirect immunofluorescence, although the antigen remains to be determined. We studied the prevalence, antigen specificity and associated clinical characteristics of anti-cytoplasmic antibodies in localized scleroderma. Serum samples from 60 patients with localized scleroderma were examined by indirect immunofluorescence analysis and immunoblotting. By immunofluorescence analysis on HEp-2 cell substrate, seven of 60 (12%) patients were shown to be positive for anti-cytoplasmic antibodies. Among these, six patients with generalized morphea had anti-mitochondrial antibodies as shown by immunoblotting: they showed reactivity with the E2 component of pyruvate dehydrogenase complex (PDC), with protein X, and with the E2 component of alpha-oxo-glutarate dehydrogenase complex, while two of them showed reactivity with PDC-E1 alpha. One of these patients who was positive for anti-PDC-E1 alpha antibody showed laboratory abnormalities, suggesting the presence of primary biliary cirrhosis. The age of disease onset was significantly higher in these six patients than in those without anti-mitochondrial antibodies. Furthermore, five of them were classified into generalized morphea with multiple plaque lesions but without linear lesions (multiple plaque type). These observations suggest that major antigens for anti-cytoplasmic antibodies in patients with localized scleroderma are mitochondrial enzymes, 2-oxo-acid dehydrogenase complexes. Patients with anti mitochondrial antibodies may comprise a unique subset of localized scleroderma designated multiple plaque type of generalized morphea of older onset. PMID- 8706338 TI - Structural polymorphisms of complement receptor 1 (CR1) in systemic lupus erythematosus (SLE) patients and normal controls of three ethnic groups. AB - CR1 exhibits a molecular weight polymorphism and variability in the number of C3b binding sites. Because this may affect immune complex clearance, we used erythrocytes to investigate the CR1 size polymorphism in SLE patients from three ethnic groups. The CR1-C allele was found more frequently in African-Americans, but the frequency did not differ between controls (10%, n = 63) and SLE patients (9%, n = 79). A 160-kD band similar to CR1-C was noted in a number of patients and was shown to be a proteolytic cleavage fragment. The study shows that the smallest form of CR1, i.e. CR1-C, is not a genetic risk factor for SLE and that the frequencies of the CR1 structural alleles do not differ from race-matched healthy controls. PMID- 8706339 TI - An analysis of apoptosis in lymphoid organs and lupus disease in murine systemic lupus erythematosus (SLE). AB - Apoptosis is a programmed cell death process that helps to regulate both T cell and B cell development. In this study, we have investigated the levels of apoptotic death in cells of the thymuses and spleens (white matter) of autoimmune MRL-lpr/lpr mice with progressive lymphadenopathy and SLE disease activity; we also examined the renal pathology in these animals. Fas is a cell surface receptor, which when activated initiates the sequence of events that lead to apoptosis. In MRL-lpr/lpr mice Fas is defective, so the competency for apoptosis may be reduced. In young animals of advancing age the thymuses enlarged until in 5-month-old females the average weight was three times that at 1 month, and spleen and kidney weights also increased in size disproportionately. At light microscope level apoptotic cells in tissue sections were counted using both routine eosin and haematoxylin staining (to identify them by their morphology) and in situ end-labelling of cells with DNA strand breaks; their presence was further confirmed by electron microscopy. As the mice aged, the numbers of apoptotic cells in thymic cortex, thymic medulla and spleen white pulp areas reduced significantly (P < 0.01-0.001), whereas in BALB/c normal controls they increased significantly (P < 0.05). These changes were coincident with the development of severe lupus, whose activity was assessed by measuring serum anti ssDNA and anti-dsDNA antibody titres and urinary protein (albumin) level which were elevated significantly by 5 months of age (P < 0.001 for both ssDNA and dsDNA and P < 0.01 for urine albumin) compared with their younger counterparts. Thus, lymphoid organ enlargement, decrease in apoptotic indices, elevated serum anti-ssDNA and anti-dsDNA antibody levels, and impaired renal function coincided with the onset and severity of lupus disease in lpr mice. It seems likely that there is a causal relationship between defective deletion of autoreactive lymphoid cells, imperfect Fas-mediated apoptosis and development of murine SLE. PMID- 8706340 TI - Diverse antigen specificity of erythrocyte-reactive monoclonal autoantibodies from NZB mice. AB - The specificities of a panel of erythrocyte-reactive MoAbs derived from NZB mice with autoimmune haemolytic anaemia (AIHA) were determined by immunoprecipitation and immunoblotting. Of the eight antibodies, two (IgG1 MoAb 105-2H and IgG2a MoAb 34-3C) immunoprecipitated a 105-kD component identified as the erythrocyte anion channel band 3. A similar band was also immunoprecipitated by the IgG2b MoAb 34 2B when used at relatively high concentrations, but none of the remaining hybridoma antibodies precipitated any labelled erythrocyte components. In immunoblotting experiments only 34-2B reacted with band 3, indicating that the epitope recognized by this MoAb is robust and differs from the determinant(s) recognized by 105-2H and 34-3C. The remaining MoAbs to react by immunoblotting were the IgM antibodies IE10 and 4C8, both of which bound to a doublet corresponding to band 4.1 from the internal erythrocyte membrane skeleton. Of the three MoAbs which gave negative results in immunoprecipitation and immunoblotting, the IgM antibodies 103-7E and 106-10E reacted poorly with intact erythrocytes by flow cytometry, but the IgG1 antibody 31-9D bound well. ELISAs demonstrated that all four IgM MoAbs are polyreactive, since they bound to histones from a panel of nuclear antigens, and additionally 103-7E reacted with phosphatidyl choline. It is concluded that band 3 is an important autoantigen in NZB AIHA. However, since 3/5 haemolytic MoAbs failed to participate this antigen, either these antibodies represent minor components of the total autoantibody response, or responses to diverse possibly non-protein surface antigens also contribute to the pathogenesis of the disease. PMID- 8706341 TI - Elastase, but not proteinase 3 (PR3), induces proteinuria associated with loss of glomerular basement membrane heparan sulphate after in vivo renal perfusion in rats. AB - Elastase, but not PR3, induces proteinuria associated with loss of glomerular basement membrane (GBM) heparan sulphate after in vivo renal perfusion in rats. PR3 and elastase are cationic neutral serine proteinases present in the azurophilic granules of polymorphonuclear leucocytes. Release of these proteolytic enzymes along the glomerular capillary wall may induce glomerular injury. Here, we investigated the effects of PR3 and elastase on the induction of proteinuria and glomerular injury after renal perfusion of these enzymes in Brown Norway rats. Perfusion of active elastase induced a dose-dependent proteinuria 24h after perfusion, while inactivated elastase did not. Perfusion of comparable amounts of active PR3 did not induce proteinuria. Light and electron microscopy showed no morphological abnormalities in any experimental group. However, immunohistology revealed that proteinuria occurring after perfusion of active elastase was associated with a strong reduction in intraglomerular expression of the heparan sulphate side chain and, to a lesser extent, of the protein core of heparan sulphate proteoglycans (HSPG). In vitro, both elastase and PR3 digested HSPG. However, PR3 bound to a lesser extent to HSPG than elastase. We conclude that elastase, but not PR3, induces proteinuria after in vivo renal perfusion. This differential effect probably relates to different binding to the GBM of those enzymes due to differences in their isoelectric points. Degradation of heparan sulfate proteoglycans, leading to the disappearance of their side chains that contribute to the polyanionic structure of the GBM, appears to be involved in the induction of proteinuria after perfusion of elastase. PMID- 8706342 TI - Immunization of non-obese diabetic (NOD) mice with glutamic acid decarboxylase derived peptide 524-543 reduces cyclophosphamide-accelerated diabetes. AB - NOD mice constitute a model for studying the prevention of human autoimmune type 1 diabetes. Glutamic acid decarboxylase (GAD) could be a key antigen involved in this disease, and GAD65 peptide 524-543 has been implicated in early T cell response in young NOD mice. We performed two i.p. injections of GAD peptide 524 543 (100 micrograms at each injection), together with Freund's incomplete adjuvant (FIA), into female NOD mice at 30 and 45 days old. Diabetes was accelerated 2 weeks later by a single injection of cyclophosphamide (CY), which acts against suppressive mechanisms. Treatment with GAD 524-543 peptide delayed the onset of diabetes and reduced its incidence (28% versus 60%; P < 0.001) compared with control mice injected with FIA alone, or GAD peptide 534-553, or an irrelevant peptide. In the same group, the severity of lymphocytic inflammation of pancreatic islets was reduced (P < 0.03). Up to 3 months after peptide injections, a strong splenocytic proliferative response occurred in immunized NOD mice against the immunizing peptide alone (but not against a panel of seven other GAD65-derived peptides). After peptide challenge of splenocytes in vitro, protection against CY-accelerated diabetes was associated with higher peptide specific production of T helper type 2 (Th2)-associated interleukins 4 and 10, whereas Th1-associated interferon-gamma and IL-2 were proportionally less represented. During contransfer, T splenocytes from GAD 524-543-immunized mice were able to reduce the capacity of T cells from diabetic donors to transfer the disease adoptively (P < 0.01), demonstrating the generation of cellular mechanisms that actively suppress the disease. It is concluded that immunization of NOD mice with GAD65 peptide 524-543 can counteract CY-accelerated diabetes, possibly through active cellular suppression linked to a shift of Th1/Th2 balance toward the production of Th2 cytokines such as IL-4 and IL-10. This study provides additional support for the notion that GAD, and more precisely its epitope 524-543, could be one of the key targets for the pathogenesis of type 1 diabetes in NOD mice, as well as for the efficacy of disease-specific peptide therapy in type 1 diabetes. PMID- 8706343 TI - Synergistic effect of deoxyspergualin (DSP) and cyclosporin A (CsA) in the prevention of spontaneous autoimmune diabetes in BB rats. AB - Dose-dependent side effects are frequently observed with immunosuppressive drugs of potential relevance for the immunotherapy of insulin-dependent diabetes mellitus (IDDM), such as CsA and DSP. If CsA and DSP acted synergistically in vivo, their combined use would allow using each compound at lower doses than those required when each drug is given in monotherapy. Consequently, dose dependent side effects could be reduced and the therapeutic activity maintained or even enforced. Toward this end we studied the effects of combined treatment with CsA and DSP on the course of IDDM in the diabetes-prone (DP)-BB rat. The results show that two 'low' doses of CsA (2 mg/kg) and DSP (1 mg/kg) that are clinically ineffective in suppressing IDDM development in BB rats when administered alone under a prolonged prophylactic regimen (30-105 days old), may successfully prevent, but not cure, the disease when given contemporaneously under the same experimental conditions. The combined treatment was well tolerated, and no side effects were noticed. These data suggest that the combined use of CsA and DSP may deserve consideration for its possible application in the prevention/treatment of human IDDM and other autoimmune diseases. PMID- 8706344 TI - Predominance of a type 2 intratumoural immune response in fresh tumour infiltrating lymphocytes from human gliomas. AB - Increasing evidence suggests the existence of polarized human T cell responses described as Th1-type (promoting cell-mediated immunity) and Th2-type (promoting humoral immunity), characterized by a dominant production of either interferon gamma (IFN-gamma) or IL-4, respectively. Little is known about the intratumoural activation of infiltrating lymphocytes (TIL) in human gliomas. Therefore, we assessed fresh TIL at cellular and molecular levels to find out if they were activated and polarized into a type 1 or 2 immune response. Flow cytometry analysis of TIL revealed that the major subset was made of T lymphocytes. Double labelling with alpha-CD3 and adhesion/ activation markers revealed T cell subsets expressing CD49a, CD49b, CD54, and CD15, some of which were almost absent in autologous T peripheral blood lymphocytes (T-PBL). Furthermore, the proportions of T-TIL expressing CD56, CD65, or CD25 were several-fold higher than in T-PBL. Intratumoural functional activation of TIL was tested by semiquantitative assessment in relative units (RU) of lymphokine gene activation with mRNA reverse transcriptase-polymerase chain reaction (RT-PCR). All TIL populations except one significantly expressed IL-4 1 to 2 logs of RU above healthy PBL baseline. Similarly, all patients expressed granulocyte-macrophage colony-stimulating factor (GM-CSF) in a range comparable to IL-4. However, most TIL populations did not express IFN-gamma, IL-2, and tumour necrosis factor-beta (TNF-beta) at higher levels than healthy normal PBL. The increase proportion of T cells expressing activation markers and the consistent detection of significant IL-4 and GM-CSF lymphokine gene activation in TIL populations suggested a predominant type 2 intratumoural immune response that does not promote cell-mediated tumouricidal activity and may contribute to the inefficiency of the antiglioma immune response. PMID- 8706345 TI - IL-12 stimulation but not B7 expression increases melanoma killing by patient cytotoxic T lymphocytes (CTL). AB - Recent studies have demonstrated that rodent tumour cells engineered to secrete a variety of cytokines, or to express foreign antigens, MHC molecules or co stimulatory molecules, are rejected by syngeneic animals. These observations have led to the initiation of a number of clinical trials using genetically modified tumour cells, to attempt to stimulate a patient anti-tumour immune response. In this study, a protocol has been developed to test in vitro the specific cytotoxic anti-tumour response generated from melanoma patient lymphocytes. The results showed that IL-12 in combination with IL-2 enhanced the autologous anti-melanoma CTL response, whereas B7.1 antigen expression on tumour cells did not increase anti-melanoma CTL generation. This method could be used to design more appropriate genetically modified tumour vaccines. PMID- 8706346 TI - Tracking of the V4-34 (VH4-21) gene in human tonsil reveals clonal isotype switch events and a highly variable degree of somatic hypermutation. AB - The V4-34 (VH4-21) gene has been found to encode certain IgM autoantibodies, and is mandatory for pathological IgM anti-erythrocyte antibodies of I/i specificity. The gene is also commonly used by normal IgM-positive B lymphocytes, but its involvement in B cells which have undergone class switching to IgG or IgA is less clear. In order to track V4-34 gene usage and class switching events during a normal immune response, we have probed RNA in a limited area of human tonsil. Results indicate that the V4-34 gene undergoes class switching to IgG or IgA, with the progeny either remaining unmutated or containing large numbers of somatic mutations. Mutational patterns indicate possible 'hot spots', and some mutations appear deleterious. At the level of individual B cells, we have tracked a clonal isotype switch event from IgM to IgA, with each retaining close to germ line configuration. In addition, we have followed a clonal switch from a mutated IgM to IgG, with no further accumulation of somatic mutations. These data indicate that the V4-34 gene is involved in a maturing immune response, and that the routes to production of IgG or IgA antibodies are various. PMID- 8706347 TI - Activation of epitope-specific memory cytotoxic T lymphocyte responses by synthetic peptides. AB - Cytotoxic T lymphocytes (CTL) recognize antigens as short peptides selected for presentation by their ability to bind to MHC class I molecules. Polyclonal Epstein-Barr virus (EBV)-specific memory CTL responses, reactivated from blood lymphocytes of HLA-A11-positive individuals by stimulation with the autologous EBV-transformed lymphoblastoid cell line (LCL), are often dominated by reactivites directed to the peptide epitope IVTDFSVIK (IVT), corresponding to amino acids 416-424 of EBV nuclear antigen-4 (EBNA4). We now report the selective activation of IVT-specific CTL by stimulation of lymphocytes with the corresponding synthetic peptide. A more than 10-fold increase in frequency of CTL clones with this specificity (from 8% to 96%) was obtained when the peptide was presented by HLA-A11-transfected T2 cells (T2/A11). Titration of synthetic peptide in cytotoxic assay demonstrated that clones activated under these conditions are as efficient as clones activated by conventional LCL stimulations. Induction of memory CTL responses required low surface density of MHC: peptide complexes, since reactivation was achieved by stimulation with T2/A11 cells pulsed with concentrations of peptide that are suboptimal for induction of target cell lysis. This protocol of activation revealed the presence of IVT-specific CTL precursors in a donor that failed to mount an IVT-specific response upon stimulation with the autologous B95.8 virus-transformed LCL. The results suggest that stimulation with synthetic peptide epitopes can be efficiently used for induction of memory CTL responses, and may be particularly helpful for the selective expansion of subdominant CTL specificities. PMID- 8706348 TI - Histaminergic regulation of interferon-gamma (IFN-gamma) production by human natural killer (NK) cells. AB - Monocytes, recovered from human peripheral blood by counter-current centrifugal elutriation, effectively inhibit the production of IFN-gamma by CD3-/56+ NK cells in response to IL-2. This study aimed at defining the nature of the inhibitory signal, particularly the importance of monocyte-derived reactive metabolites of oxygen. It was found that monocytes recovered from patients with chronic granulomatous disease (CGD), a condition characterized by deficient NADPH-oxidase activity of phagocytes, did not inhibit IFN-gamma production by NK cells. Further, catalase, a scavenger of hydrogen peroxide, completely reversed the inhibitory signal whereas scavengers of the superoxide anion, hypohalous acids, the hydroxyl radical, or nitric oxide synthesis inhibitors such as L-NMMA were ineffective. Inhibition of IFN-gamma production was operating on a pretranslational level, as indicated by the inability of enriched NK cells to accumulate IFN-gamma mRNA in the presence of elutriated monocytes. Hydrogen peroxide, at micromolar concentrations, reconstituted the inhibition of IFN-gamma production when added to enriched NK cells. Histamine, a biogenic amine which inhibits the generation of reactive oxygen metabolites in monocytes, abrogated the inhibition of IFN-gamma production in NK cells; by this mechanism, histamine strongly synergized with IL-2 to induce IFN-gamma in mixtures of NK cells and monocytes. The synergizing effect of histamine was specifically mediated by H2 type histamine receptors. We conclude that: (i) the induction of IFN-gamma mRNA in NK cells is effectively down-regulated by products of the oxidative metabolism of monocytes; and (ii) histamine effectively enhances IFN-gamma production by preventing monocyte-induced oxidative damage to NK cells. PMID- 8706349 TI - Lysis of pulmonary fibroblasts by lymphokine (IL-2)-activated killer cells--a mechanism affecting the human lung microenvironment? AB - In this study we investigated whether IL-2-activated killer cells may bind and exert lytic activity against non-transformed lung fibroblasts. We demonstrated that human lymphokine-activated killer (LAK) cells generated in vitro following incubation with recombinant IL-2 of either peripheral blood mononuclear cells (PB LAK) or lymphocytes obtained from bronchoalveolar lavage (BAL-LAK), but not resting cells, can lyse normal lung fibroblasts obtained from transbronchial lung biopsies in a 4-h 51Cr release assay. Both autologous and allogeneic fibroblasts were consistently lysed by LAK cells, thus suggesting that the phenomenon we observed is not MHC-restricted. Since fibroblasts can bind IL-2 through specific receptors, we evaluated whether long-term culture with rIL-2 could modulate the susceptibility to lysis of target cells. Our data showed that autologous fibroblasts were more resistant to lysis than allogeneic fibroblasts when they were cultured with rIL-2. Since LAK cells have been demonstrated to release a series of different immunomodulatory cytokines, we evaluated the effect of short term incubation of fibroblasts with different factors, including IL-1, IL-2, IL 3, IL-4, IL-6, tumour necrosis factor-alpha (TNF-alpha), and interferon-gamma (IFN-gamma), on the binding and the lysis mediated by LAK cells. These cytokines were not directly cytotoxic on fibroblasts. Only IFN-gamma was found to have a significant protective effect against the lysis. Our data support the concept that a self-directed cytotoxicity against pulmonary fibroblasts is generated during lymphocyte activation with rIL-2. PMID- 8706351 TI - Gaining a competitive edge: how to present yourself at your personal best. PMID- 8706350 TI - Effects of ultraviolet B irradiation on cell-cell interaction; implication of morphological changes and actin filaments in irradiated cells. AB - We studied the effects of ultraviolet B (UV-B) irradiation on cell-cell interactions using mouse lymphoma RMA cells and T cell hybridoma HTB-176.10. RMA cells act as stimulators by presenting H-2Kb surface antigens to HTB-176.10 cells, inducing IL-2 production in HTB-176.10 cells. Irradiating RMA cells with 1000J/m2 UV-B suppressed cell cluster formation between RMA and HTB-176.10 cells and reduced the level of IL-2 production in HTB-176.10 cells, although H-2Kb surface antigens of RMA cells were still expressed. Electron microscopic observations of irradiated RMA cells revealed that UV-B irradiation damaged cell structures, resulting in the disappearance of microvilli on the cell surface, destruction of mitochondria, vacuolation of cytoplasm and swelling of the perinuclear cisterna space. We found that these alterations were accompanied by polymerization of filamentous actin quantified by flow cytometry after NBD phallacidin staining. Our results suggest that a target of UV-B-induced alterations is actin filaments, which support the cell morphology as the cytoskeleton, and that modification of filamentous actin inhibits interaction between RMA and HTB-176.10 cells. This underlying mechanism may account for the impaired interaction between antigen-presenting cells and T cells after transfusion with UV-B-irradiated allogeneic blood components. PMID- 8706352 TI - Gaining a competitive edge: how to present yourself at your personal best. PMID- 8706353 TI - C4 phenotypes in IgA nephropathy: disease progression associated with C4A deficiency but not with C4 isotype concentrations. AB - IgA nephropathy (IgAN) is a common glomerular disease and is thought to have an immunological origin which may involve complement-mediated pathogenic mechanisms. We performed C4 phenotyping and C4 isotype quantification in 93 IgAN patients in Southern Sweden. Phenotype frequencies did not deviate from those of healthy controls. However, three patients had homozygous C4A deficiency and these all belonged to a group of fifteen patients with end-stage renal failure (p < 0.0035). Progression to end-stage renal failure did not appear to be faster than in other IgAN patients. Both C4A and C4B concentrations were raised in the IgAN patients, but the C4A/C4B concentration ratios did not deviate from those of healthy controls. This indicated that heterozygosity for C4A or C4B deficiency or other reasons for the relatively low concentrations of the protein were not associated with disease susceptibility. There was no correlation between low C4A/C4B ratio and poor prognosis. In conclusion, the findings suggested that homozygous C4A deficiency predisposes to development of end-stage renal failure. The question if this is due to complement dysfunction or to linked genetic factors remains to be elucidated. PMID- 8706354 TI - Favorable prognostic significance of raised serum C3 concentration in patients with idiopathic focal glomerulosclerosis. AB - This study examined the relationship between the clinical and pathologic variables and renal outcome in patients with idiopathic focal glomerulosclerosis (FGS). The study population included 106 patients with biopsy proven idiopathic FGS followed for a mean of 31 months. Forty-nine patients (46%) maintained stable renal function, 21 patients (20%) had a progressive deterioration of renal function and an additional 36 patients (34%) progressed to end-stage kidney disease. By multivariate analysis the following baseline variables (i.e. variables measured at the time of renal biopsy) were associated with the loss of renal function: 1. An elevated serum creatinine; 2. an elevated systemic blood pressure; 3. extensive interstitial fibrosis and tubular atrophy by kidney biopsy; 4. a low-normal serum C3 level. At baseline serum C3 levels were abnormally high in 31 patients (29%). Baseline C3 levels were inversely correlated with baseline serum creatinine levels (p < 0.0001), the severity of histologic changes (p < 0.0001), and better preservation of renal function during follow-up (p < 0.0001). Serum C3 levels did not correlate with proteinuria or with other clinical variables tested. The presence of C1q in the mesangium did not correlate with renal outcome. In conclusion, serum C3 levels measured at the time of biopsy correlate with renal outcome in patients with idiopathic FGS. The basis for this correlation is unknown but it cannot be fully explained by relationships between serum C3 levels and other clinical variables. PMID- 8706355 TI - The clinico-pathological characteristics and outcome in hemolytic-uremic syndrome of adults. AB - Twenty-eight adults who developed hemolytic-uremic syndrome (HUS) underwent a renal biopsy during the period from 1973 to 1993, and the specimens were analyzed both clinically and pathologically. The observed etiologic antecedents of HUS were infectious disease in 7 cases, medication, mainly due to cytotoxic drugs in 7 cases, post-partum in 7 cases, progressive systematic scleroderma in 1 case and unknown in 6 cases. Histologically, 17 cases were glomerular type, 3 cases were arterial type, 6 cases were mixed type and 2 cases demonstrated cortical necrosis. The preceding conditions and the histological findings appeared not to be related to each other. In the acute phase of the disease, of 11 patients who fell into severe acute renal failure (ARF) with an increase in the serum creatinine level of more than 5 mg/dl, 8 patients needed dialysis therapy while 4 patients did not recover from the disease (3 died and 1 was introduced to maintenance dialysis therapy). The incidence of thrombus in the glomerular capillaries and fibrinoid necrosis of the arterioles, the index of the expansion of glomerular subendothelial lucent zone (Glom-SE-ex), the index of glomerular sclerosing and the index of the intimal thickening of the arterioles were higher in the severe ARF than in the mild ARF. After analyzing the prognostic factors on renal survival, severe ARF, the presence of fibrinoid necrosis, severe glomerular endothelial damage and severe intimal thickening of the arterioles appeared to have a significantly adverse effect. Moreover, high Glom-SE-ex and the presence of fibrinoid necrosis also independently influenced renal survival. We thus conclude that the etiology of HUS in adults demonstrates a greater variation than that in children. Therefore, it is very difficult to predict the outcome of this disease based on the clinical findings alone, and thus a renal histological examination is considered to help in determining the long-term outcome. PMID- 8706356 TI - Effects of dietary sucrose and fibers on blood pressure in hypertensive rats. AB - In spontaneously hypertensive rats (SHR), two separate studies examined effects on systolic blood pressure (SBP) and other cardiovascular parameters of different concentrations of sucrose compared to starches, soluble fibers (guar, psyllium), and insoluble fibers (cellulose, wheat bran). In the initial study, four diets were tested. The first diet was relatively high in sucrose calories (50%) and low in protein calories (17%)--"high sucrose"; the second diet was relatively low in sucrose (11%) and high in protein (56%) calories--"low sucrose". The third and fourth diets resembled the first and second diets respectively, but cornstarch replaced sucrose--"high and low starch". Initial SBP in each group averaged approximately 168 mmHg. After 2 weeks of ingesting the special diets, SBP of the high sucrose group rose rapidly and significantly, eventually rising above 200 mmHg by the termination of examination. The other 3 groups maintained the original SBP until after the 3rd week when the low sucrose group developed a rapid and significant SBP elevation approaching 200 mmHg. SBP of high starch and low starch remained below 181 mmHg for the two months of study. Platelets obtained at the termination of the study from the sucrose groups compared to the starch groups showed increased aggregatory responses to collagen and ADP. Further, significant elevations of triglycerides and cholesterol in the high sucrose group were found. The former parameter was also significantly elevated in the low sucrose group. In the second study, adding guar and psyllium to high sucrose diets delayed sugar-induced hypertension, while cellulose and wheat bran virtually showed no effects. Serum insulin concentrations correlated positively with SBP, serum triglycerides, and glucose--not cholesterol. Accordingly, sucrose compared to starch ingestion in SHR can adversely influence SBP and various other cardiovascular risk factors. These effects can be delayed by the presence of soluble fibers, but not insoluble fibers, in the diets. PMID- 8706357 TI - Taste acuity in patients with chronic renal failure. AB - Taste acuity for the four primary tastes has previously been shown to be impaired in patients with chronic renal failure (CRF) and maintenance dialysis treatment, although data on CAPD (continuous ambulatory peritoneal dialysis) patients are insufficient. In a test group of 57 CRF patients and 57 healthy controls, matched for age, sex and body mass index (BMI, kg/m2) taste acuity for the four primary tastes was determined. Fourteen patients were on CAPD treatment and 12 patients on hemodialysis (HD). No patients or controls were diabetic, on antibiotic treatment or had a malignant disease. Taste tests were standardized and performed on dialysis-free days for HD-patients. Taste acuity for bitter and salt was significantly lower for preuremic patients compared to their controls. In CAPD patients taste detection of bitter was impaired and in HD-patients detection of salty taste was impaired. In conclusion taste acuity is impaired in uremic patients and dialysis patients including CAPD. The mechanism for taste alterations remain to be explained. PMID- 8706358 TI - Safety of pamidronate in patients with renal failure and hypercalcemia. AB - Pamidronate (APD) is a drug widely used for the treatment of hypercalcemia of malignancy. Renal impairment has been associated with the use of other bisphosphonates in humans, and nephrotoxicity has been described after APD administration in animals. We retrospectively evaluated the safety and efficacy of APD administration in 31 patients with underlying renal insufficiency who received 33 courses of APD in doses of 60-90 mg. Hypercalcemia resolved or improved in 91% of the patients and only 1 case had severe hypocalcemia. A transient deterioration in renal function was observed in 8 courses but this was unrelated to APD administration. No systemic ill effects were observed. APD appears to be a safe drug in patients with underlying renal failure. PMID- 8706359 TI - Clinical experience with a new recombinant hepatitis-B vaccine in previous non responders with chronic renal insufficiency. AB - Patients with renal insufficiency have an increased risk of hepatitis B infection and a high probability to develop a chronic course of this disease. After hepatitis B vaccination they are known to show a low rate of seroconversion. In the present study we assessed the efficacy of a new recombinant pre-S1 and pre-S2 containing hepatitis B vaccine in 17 non-responders (anti-HBs titer 0) and 4 low responders (anti-HBs titer < or = 5 IU/ml) with chronic renal insufficiency (16 on chronic hemodialysis therapy and 5 without hemodialysis treatment). Seroconversion rate was 65% after the third and 71% after the fourth vaccination. Only minor side effects were seen. These results encourage to use the new vaccine in a larger number of patients with renal insufficiency. PMID- 8706360 TI - Improved urea clearance raises the BUN in continuous peritoneal dialysis. AB - Cross-sectional studies in steady state dialysed chronic end-stage renal failure patients show urea clearance (Kt/V) and total urea excretion (protein catabolic rate) correlate positively. However, urea clearance is total urea excretion divided by BUN. Thus urea clearance and BUN relate reciprocally, and so their mathematical product (total urea excretion) is independent of clearance. As such clearance cannot also be a positive correlate of total excretion as demanded by the cross-sectional studies. Furthermore the clearance formula dictates that the positive urea clearance and total urea excretion correlation found in the cross sectional studies can only occur if the increased urea clearance fails to reciprocally lower the BUN. Thus the relations of urea clearance, urea excretion, and BUN requires further definition. To so define we examine dialysis urea excretion, dialysis urea clearance, BUN, and serum albumin in 13 stabilized chronic uremics with minimal native renal function who are treated by continuous ambulatory peritoneal dialysis (CAPD). Urea clearance and BUN correlate positively (r = 0.62, p < 0.05) and both also correlate positively with dialytic urea excretion and (urea clearance r = 0.912, p < 0.001, BUN r = 0.88, p < 0.001). In addition dialytic urea excretion and serum albumin indexed to body size correlate positively (p < 0.05). Thus in the steady state urea clearance associates with both an increase in BUN and urea output. However the law of conservation of mass makes urea output is a function of protein intake. Thus increased clearance cannot directly increase such output, and so increased clearance must first increase intake but in doing so it increases the retention of the byproducts of enhanced intake, BUN and other protein metabolites, so leading to a paradox, the more removed, the more remains. These observations taken together suggest that in chronic uremia treated by continuous dialysis, elevation of the BUN may be a marker for an adequate restoration of protein metabolism if inadequate dialysis is excluded. PMID- 8706361 TI - The effect of ultra-low sodium dialysate in CAPD. A kinetic and clinical analysis. AB - In peritoneal dialysis, sodium (Na) passes through the peritoneal membrane to a lesser extent than H2O and thus hypotonic fluid is removed indicating that Na removal is inefficient. Therefore we developed an ultra-low Na dialysate (ULNaD) [Na 98 mmol/liter, osmolality 340 mOsm/kg, glucose (G), 2.48%] to facilitate trans-peritoneal Na removal. The aim of the study was to examine (a) the fluid and Na kinetics of this solution, and (b) the clinical effect in the overhydrated patient. Regarding the former, in six cases, changes in the effluent Na levels during a 240-minute dwell of ULNaD were measured at 60-minute intervals. In eighteen cases, the transperitoneal net-Na removal (Na removal) and drained ultrafiltration (UF) volume after a 240-minute dwell of ULNaD were measured and compared to two types of standard Na dialysate (SNaD) [Na 132 mmol/liter, 1.36% G (348 mOsm/kg) and 2.27% G (396 mOsm/kg)]. The effluent Na levels increased gradually until the end of the dwelling. Drained UF volume of ULNaD (224 +/- 25 ml) was more than 1.36% G SNaD (77 +/- 33 ml, p < 0.01), but it was less than 2.27% G SNaD (341 +/- 32 ml, p < 0.01). However, a significant increase in Na removal by ULNaD (78.1 +/- 5.6 mmol) was found compared with both SNaD solutions (12.6 +/- 6.4 mmol in 1.36% G solution, p < 0.001; 27.8 +/- 5.1 mmol in 2.27% G solution, p < 0.001, respectively). Regarding the latter study aim, in six overhydrated patients with a Na excess, SNaD was replaced with ULNaD once a day for 7 consecutive days. At the end of the study as compared to the pre-values, a significant reduction in body weight (-2.5 +/- 1.0%, p < 0.05) and mean arterial pressure (-12.3 +/- 3.5%, p < 0.001) was observed. An increased drainage of daily UF volume (+86.5 +/- 31.1%, p < 0.05) and a markedly increased daily Na removal (+131.1 +/- 24.2%, p < 0.01) were also observed. The results of this study suggest that ULNaD actually facilitates trans-peritoneal Na removal mainly by diffusion force, and that ULNaD could benefit overhydrated patients with a Na excess. PMID- 8706362 TI - Impact of peritoneal absorption of glucose on appetite, protein catabolism and survival in CAPD patients. AB - Dietary protein and calorie intake, protein catabolism and peritoneal kinetics were measured in 97 CAPD patients to establish the effect of peritoneal glucose absorption on appetite and survival. There was a large variability in the number of calories obtained from the dialysate, mean 5.89 cal/kg (median 5.43 cal/kg), with a skewed distribution, due to the increased requirement for hypertonic solutions by patients with more rapid glucose absorption and poor ultrafiltration. On average calories derived from peritoneal absorption accounted for 19% of the total energy intake which in itself was well below that recommended. Patients with > 6 cal/kg, obtained from the dialysate (top 20th percentile, n = 19) were compared with those with < 6 cal/kg, but no significant differences in oral protein or calorie intake, protein catabolism or total calorie intake were found. Age, body mass index (BMI) and KT/V were also similar in both groups. Patients were followed-up prospectively for a minimum of 24 months and a comparison made of actuarial survival. Patients with high peritoneal calorie intake tended to survive longer but this was not significantly different (p = 0.25). This study suggests that calories derived from the peritoneum in CAPD patients do not suppress appetite, provide a useful and significant proportion of the total energy intake, that does not cause excessive obesity or have a negative effect on patient survival. PMID- 8706363 TI - Assessment of health status in peritoneal dialysis patients: a potential outcome measure. AB - OBJECTIVES: To determine the feasibility and practicality of measuring general health status (GHS) in an outpatient peritoneal dialysis population. To determine whether GSH correlated intuitively with biochemical, socio-demographic and co morbidity measurements. DESIGN: The Medical Outcomes Study 20-item short form was administered on a voluntary basis in the outpatient setting. Demographic and current biochemical data were extracted from the medical record. The effects of the socio-demographic, biochemical and physiologic variables on the six subscales of GHS generated by the questionnaire were estimated using multivariate linear regression analysis resulting in the development of six separate models. SETTING: Peritoneal dialysis program of a University Hospital. PATIENTS: Sixty stable patients on home peritoneal dialysis completed the GHS questionnaire during regularly scheduled outpatient visits. Ages ranged from 13 to 81 years. The study group included 14 diabetics (23%). RESULTS: Administering the questionnaire caused no logistical difficulties in the outpatient setting. Regression models for predicting GHS were both significant and intuitively correct. The presence of co-morbidities such as diabetes mellitus (p = 0.002; Social Subscale) and peripheral vascular disease (p = 0.016: General Health Subscale) had the most significant negative impact on GHS. An increased length of time on dialysis was associated with a higher GHS (p = 0.002; Physical Subscale). CONCLUSION: General Health Status questionnaires can be readily administered to peritoneal dialysis patients in the outpatient setting. They have face validity as a measurement of wellness and functioning. The longitudinal use of such instruments in conjunction with clinical and laboratory findings may identify both medical and non-medical factors impacting on our peritoneal dialysis population. PMID- 8706364 TI - Obstructive uropathy due to interstitial cystitis in a patient with systemic lupus erythematosus. AB - A patient with documented SLE is described who presented with dysuria, gastrointestinal (GI) symptoms and renal insufficiency associated with the unusual occurrence of bilateral hydro-ureteronephrosis due to ureterovesical junction stricture (obstructive uropathy) on radiographic findings. Pathologic investigations disclosed chronic interstitial cystitis (IC) with evidence of focal immune complex deposition in blood vessel walls of the bladder. The GI symptoms and dysuria regressed with initial therapy of SLE by steroid but persistent obstructive uropathy (OU) and renal insufficiency required bilateral nephrostomy. Then, following continuous medical treatment with steroid plus intravenous pulse injection of cyclophosphamide, obstructive uropathy was relieved even with removing nephrostomy tube and renal function remained stable. PMID- 8706365 TI - Rapidly progressive crescentic glomerulonephritis and Legionnaires' disease. PMID- 8706366 TI - Pharmacologic treatment of congestive heart failure. AB - Congestive heart failure is a clinical syndrome producing symptomatic deterioration, functional impairment, and shortened life span. The syndrome is complex in that it includes both peripheral and cardiac effects which contribute to the progression of heart failure. In the periphery, elevations in the sympathetic nervous system and renin-angiotensin system increase afterload and contribute to further salt and water retention. The central cardiac abnormalities include remodeling of the heart and downregulation of beta receptors. Traditional heart failure therapy has included treatment of fluid retention with diuretics, although their effect on mortality has never been addressed. The most proven therapy in heart failure is treatment with vasodilators, particularly angiotensin converting enzyme (ACE) inhibitors. Improved survival with ACE-inhibitor therapy has been demonstrated in patients with severe heart failure (CONSENSUS), mild to moderate heart failure (SOLVD), and in comparison with vasodilator therapy with hydralazine isosorbide dinitrate (VHeFT II). Improved survival has also been noted in postmyocardial infarction when the ejection fraction is decreased (SAVE). The ACE inhibitors have now become standard therapy for heart failure regardless of severity. Additive vasodilator therapy with calcium-channel antagonists is under investigation. Inotropic therapy is controversial at present because of disappointing mortality results. The clinical mainstay digitalis remains without convincing mortality reduction data. Other inotropic agents, particularly phosphodiesterase inhibitors, have shown uniformly negative survival results. However, the new mixed action agents vesnarinone and pimobenden have shown favorable data, with vesnarinone demonstrating a mortality reduction effect. Beta-blocker therapy in heart failure has also found renewed interest, particularly with the new agents carvedolol and bucindolol which also have vasodilating properties. PMID- 8706367 TI - Newer Doppler measures of left ventricular diastolic function. AB - Left ventricular (LV) diastolic dysfunction is an important cause of heart failure, and recent advances in the application of Doppler techniques allow a semiquantitative assessment of LV diastolic performance. This review discusses the use of Doppler echocardiography in the comprehensive assessment of LV diastolic function and performance in terms of the normal mitral and pulmonary venous flow profiles, their physiologic basis, and alterations in diseased states. There is also a discussion on the newer aspects of mitral flows such as relative durations of mitral A and pulmonary vein AR waves, E- and A- wave propagation inside the LV with their hemodynamic correlates, and derivation of ventricular dP/dt and Tau from the mitral regurgitation velocity profile. Analysis of these flow profiles and the other Doppler measures alluded to above allow one to make a fairly precise hemodynamic assessment of a patient in terms of left atrial pressure, LV relaxation and stiffness and the profile of LV diastolic pressure in terms of pre- 'a' wave and 'a' wave pressures and ventricular end-diastolic pressure. PMID- 8706368 TI - Evidence for a relation between inspired gas mixture and the left ventricular contrast achieved with Albunex in a canine model. AB - BACKGROUND: In a previous experiment, a marked reduction in the right- and left sided contrast effect of Albunex was noted in an intubated animal spontaneously breathing isoflurane in 100% oxygen. The theory suggests that the time course of echogenicity of microbubbles in liquid is dependent on the pressure and the gradients of dissolved gases. The present set of experiments tested whether the loss of contrast occurs at commonly used therapeutic concentrations of inspired oxygen. HYPOTHESIS: This research tested the hypothesis that the left ventricular (LV) contrast effect achieved with intravenous injection of the ultrasound contrast agent Albunex is related to the inspired oxygen content. METHODS: Intubated dogs were maintained in a spontaneously respiring anesthetic state on isoflurane and mixtures of oxygen (12-50%) in nitrogen. FIO2 was held steady for 15 min prior to injection of 0.08 ml/kg of Albunex. The contrast effects were recorded from a transthoracic short-axis view. Left and right ventricular brightness curves were generated from digitized sequences of end-diastolic frames. The minimum and maximum brightness and area under the time-brightness curves were determined. RESULTS: The LV maximum brightness and area under the curve showed significant negative correlations (p = < 0.004) with the FIO2, while the minimum brightness showed a significant positive correlation (p = < 0.002). No significant correlations were found for the right ventricular brightness parameters. CONCLUSIONS: These findings show an important relationship between the FIO2 and loss of the contrast effect of Albunex. This loss occurs at oxygen concentrations in the therapeutic range, but could be overcome by increasing the dose of Albunex. The mechanism is likely related to an outward nitrogen gradient causing a loss of echogenicity. The clinical implication is that patients on supplemental oxygen may require higher doses of Albunex to achieve optimal opacification. PMID- 8706369 TI - Effects of heart rate on flow velocity of the left atrial appendage in patients with nonvalvular atrial fibrillation. AB - BACKGROUND AND HYPOTHESIS: Flow velocity of the left atrial appendage (LAA) is thought to be important in thrombus formation in association with blood stasis and the development of spontaneous echo contrast. The effects of heart rate on peak flow velocity of LAA have not been studied in patients with nonvalvular atrial fibrillation. METHODS: Using transesophageal Doppler echocardiography, peak flow velocity of the LAA was measured at the junction between the left atrium and the LAA during left ventricular (LV) systole and diastole in 21 patients with nonvalvular atrial fibrillation. In six cases, the average peak flow velocity of the LAA for 10 consecutive beats with moderately long R-R intervals (LI beats) was compared with those of 3-5 consecutive beats with extremely short R-R intervals (SI beats). RESULTS: Average peak flow velocity of the LAA during LV diastole was significantly higher than that during LV systole (26.5 +/- 15.7 vs. 19.3 +/- 10.4 cm/s, p < 0.01). In SI beats, average peak flow velocity of the LAA was significantly lower than that in LI beats (17.1 +/- 12.1 vs. 21.2 +/- 12.9 cm/s, p < 0.01). CONCLUSION: An increased heart rate reduced the peak flow velocity of the LAA in patients with nonvalvular atrial fibrillation, which would promote blood stasis in the LAA. PMID- 8706370 TI - Association of conventional and exertional coronary heart disease risk factors in 5,000 apparently healthy men. AB - BACKGROUND AND HYPOTHESIS: The aim of this study was to define the prevalence of previously undetected coronary heart disease among asymptomatic males, aged 30-65 years, by means of resting and exercise electrocardiography in conjunction with an analysis of conventional and exertional coronary risk factors. METHODS: Between January 1985 and December 1989 we examined 5,000 clinically asymptomatic subjects. A detailed case history was obtained for each individual, followed by a complete physical examination, comprehensive blood (including lipid) profile, lung function tests, chest x-ray, a resting 12-lead electrocardiogram (ECG), and a maximal treadmill exercise ECG. Whenever possible, on-line computerized respiratory analysis (Beckman Metabolic Measurement Cart) was carried out during the exercise tests. Conventional and exertional coronary heart disease risk factors were also recorded. RESULTS: A total of 162 persons (3.2%) showed abnormal S-T segment responses during the exercise or recovery period. Of these, 92 subjects underwent further investigations: coronary angiography (79), 201thallium scanning (13), 201thallium scanning followed by coronary angiography (7). Of the 86 patients who proceeded to coronary angiography, 19 (22%) had either normal coronary artery anatomy or only insignificant disease. Among the 67 (78%) of patients with significant angiographically demonstrable disease, 26 received coronary artery bypass grafting, 7 underwent coronary angioplasty, and the remainder continued on medical management. CONCLUSIONS: These results are discussed in relation to a variety of conventional and exertional coronary risk factors. PMID- 8706371 TI - F-amplitude, left atrial appendage velocity, and thromboembolic risk in nonrheumatic atrial fibrillation. Stroke Prevention in Atrial Fibrillation Investigators. AB - BACKGROUND: Reduced left atrial appendage velocity (LAAV) has been identified as a marker for thromboembolism in patients with atrial fibrillation. HYPOTHESIS: It was postulated that electrocardiographic (ECG) F-wave amplitude would correlate with LAAV, and inversely with the risk of thromboembolism in patients with atrial fibrillation. METHODS: In all, 53 patients with nonrheumatic (NRAF) and 7 patients with rheumatic (RAF) atrial fibrillation underwent assessment of maximum LAAV, which was correlated to the maximum ECG F-wave voltage from lead V1 (F(max)). In 450 NRAF patients on neither aspirin nor warfarin, the relationship between F(max) and thromboembolic risk was assessed over an average follow-up of 1.3 years. RESULTS: F(max) did not correlate with LAAV (r = 0.2, p = 0.07). Patients with intermittent atrial fibrillation (n = 123) had smaller F(max) amplitude than patients with constant atrial fibrillation (n = 327) (mean 0.73 vs. 0.88 mV-1, p = 0.001). F(max) amplitude was not related to a history of hypertension, systolic blood pressure, duration of NRAF, abnormal transthoracic echocardiographic left ventricular (LV) systolic function or left atrial (LA) diameter. There was a strong trend for increased LV mass being related to smaller F(max) amplitude after adjusting for body surface area (p = 0.06). F(max) amplitude was not correlated with risk of embolic events, including only those events presumed by a panel of case-blinded neurologists to be cardioembolic. CONCLUSIONS: F(max) amplitude in NRAF is smaller in patients with intermittent versus constant AF. It does not correlate with LAAV, LA size, increased LV mass, or systolic dysfunction, hypertension, or risk of embolism. Therefore, F(max) amplitude may not be used as a surrogate for LAAV, or as a measure of thromboembolic risk in NRAF. PMID- 8706372 TI - Holmium laser-assisted coronary angioplasty in acute ischemic syndromes. AB - BACKGROUND AND HYPOTHESIS: Initial studies have shown holmium laser to be effective in ablation of coronary atheroma, and small studies that it may be helpful in ablation of thrombotic stenoses. Therefore, holmium laser-assisted coronary angioplasty was studied in 85 consecutive patients with acute ischemia syndromes. METHODS: Indications for therapy were acute myocardial infarction (MI) in 7 patients (8%), post-MI ischemic in 32 patients (38%), and crescendo angina pectoris in 46 patients (54%). Coronary morphology characteristics by multivessel angioplasty prognosis group criteria were Type A in 9 (10%), Type B1 in 15 (18%), Type B2 in 44 (52%), Type C in 17 patients (20%). RESULTS: Angiographic evidence of thrombus was seen in 37 (44%) of patients. The laser successfully crossed the total length of the coronary narrowing in 76 patients (89%). Procedure/clinical success was 92% for the total study population, 100% for patients with acute MI, 94% for post-MI ischemia patients, and 89% for patients with crescendo angina. Lesions with and without thrombus had identical procedure success rates. Major complication rate was 3.5%, (deaths 0%, Q-wave MI 0%, and emergent bypass surgery 3.5%). Six-month angiographic restenosis rate (> 50% stenosis) was 45%. CONCLUSION: Holmium laser-assisted balloon angioplasty appears promising in the treatment of acute ischemic syndromes and thrombotic coronary lesions. PMID- 8706373 TI - Prognosis of patients with frequent premature ventricular complexes and nonsustained ventricular tachycardia after coronary artery bypass graft surgery. AB - BACKGROUND: Previous studies in small groups of predominantly nongeriatric patients showed that complex ventricular arrhythmias occurring after coronary artery graft (CABG) surgery are of no prognostic significance. The purpose of this study was to compare the prognosis of patients with and without advanced grade ventricular arrhythmias (AGVA) after CABG in a large group of patients. [In this paper, AGVA is used as an abridged definition of frequent premature ventricular complexes (PVCs) and nonsustained ventricular tachycardia (NSVT) which represent advanced grade ventricular arrhythmias.] METHODS: Twenty-four hour ambulatory electrocardiographic (ECG) monitoring was performed 3 days after CABG in 185 consecutive patients with 185 closely matched control patients without AGVA. Of 185 patients with AGVA, 77 had frequent PVCs, 45 had NSVT, and 63 patients had both. The average age of both groups was 65 +/ 9.7 years. Patients were followed for 34 +/ 10 months, and in 30 patients ambulatory monitoring was repeated at the end of the follow-up. RESULTS: Fifteen AGVA and nine control patients died. In each group seven deaths were noncardiac. Six nonsudden and two sudden cardiac deaths (SCD) occurred in the AGVA group at 2-36 months after CABG and two nonsudden cardiac deaths in the control group at 3 and 35 months after CABG (p = 0.053). Both SCDs occurred 33 months after CABG after new events known to predispose to SCD. In 18 of 30 patients AGVA was no longer present when ambulatory ECG monitoring was repeated 36 +/ 11 months after CABG. CONCLUSION: AGVA after CABG was not a marker of an early sudden cardiac death. In 60% of patients not treated with antiarrhythmic drugs, AGVA was no longer present late after operation. PMID- 8706374 TI - Ionic mechanisms of ischemia-related ventricular arrhythmias. AB - The aim of this review is the utmost simplification of the cellular electrophysiologic background of ischemia-related arrhythmias. In the acute and subacute phase of myocardial infarction, arrhythmias can be caused by an abnormal impulse generation, abnormal automaticity or triggered activity caused by early or delayed afterdepolarizations (EAD and DAD), or by abnormalities of impulse conduction (i.e., reentry). This paper addresses therapeutic intervention aimed at preventing the depolarization of "pathologic" slow fibers, counteracting the inward calcium (Ca) influx that takes place through the L-type channels (Ca antagonists), or hyperpolarizing the diastolic membrane action potential, increasing potassium (K) efflux (K-channel openers) in arrhythmias generated by an abnormal automaticity (ectopic tachycardias or accelerated idioventricular rhythms). If the cause enhanced impulse generation is related to triggered activity, and since both EAD and DAD are dependent on calcium currents that can appear during a delayed repolarization, the therapeutic options are to shorten the repolarization phase through K-channel openers or Ca antagonists, or to suppress the inward currents directly responsible for the afterdepolarization with Ca blockers. Magnesium seems to represent a reasonable choice, as it is able to shorten the action potential duration and to function as a Ca antagonist. Abnormalities of impulse conduction (re-entry) account for the remainder of arrhythmias that occur in the acute and subacute phase of ischemia and for most dysrhythmias that develop during the chronic phase. Reentrant circuits due to ischemia are usually Na channel-dependent. Drug choice will depend on the length of the excitable gap: in case of a short gap (ventricular fibrillation, polymorphic ventricular tachycardia, etc.), the refractory period has been identified as the most vulnerable parameter, and therefore a correct therapeutic approach will be based on drugs able to prolong the effective refractory period (K-channel blockers, such as class III antiarrhythmic drugs); on the other hand, for those arrhythmias characterized by a long excitable gap (most of the monomorphic ventricular tachycardias), the most appropriate therapeutic intervention consists of depressing ventricular excit-ability and conduction by use of sodium-channel blockers such as mexiletine and lidocaine. Compared with other class I antiarrhythmic agents, these drugs minimally affect refractoriness and exhibit a use-dependent effect and a voltage dependent action (i.e., more pronounced on the ischemic tissue because of its partial depolarization). PMID- 8706375 TI - Regional cardiac compression due to a large anaerobic bacterial empyema. AB - Compression of the heart outside the pericardial sac is a rare cause of hemodynamic compromise and cardiac tamponade. We report an atypical case of regional cardiac compression caused by a large loculated anaerobic bacterial empyema. PMID- 8706376 TI - Malposition of transvenous pacing lead in the left ventricle. AB - Transvenous pacemaker lead malposition in the left ventricle occurs rarely and requires a high index of suspicion for proper diagnosis. The case of a woman with unintentional lead placement in the left ventricle is presented. She had two episodes of transient neurologic deficits, possible secondary to embolic events, and was started on oral anticoagulants. Chest x-ray and electrocardiogram (ECG) suggested pacemaker lead malposition and transesophageal echocardiography revealed sinus venosus atrial septal defect. The lead was shown to cross the atrial septum and the mitral valve to the left ventricle. The malpositioned lead was successfully removed from the left ventricle at the time of surgical repair of the atrial septal defect. The potential value of 12-lead ECG, chest x-ray, posteroanterior and lateral views, and echocardiography in the diagnosis of pacemaker lead malposition are discussed and recommendations to avoid this complication at the time of pacemaker implant are outlined. PMID- 8706377 TI - Etienne-Jules Marey: 19th century cardiovascular physiologist and inventor of cinematography. PMID- 8706378 TI - Essentialism, word use, and concepts. AB - The essentialist approach to word meaning has been used to undermine the fundamental assumptions of the cognitive psychology of concepts. Essentialism assumes that a word refers to a natural kind category in virtue of category members possessing essential properties. In support of this thesis, Kripke and Putnam deploy various intuitions concerning word use under circumstances in which discoveries about natural kinds are made. Although some studies employing counterfactual discoveries and related transformations appear to vindicate essentialism, we argue that the intuitions have not been investigated exhaustively. In particular, we argue that discoveries concerning the essential properties of whole categories (rather than simply of particular category members) are critical to the essentialist intuitions. The studies reported here examine such discovery contexts, and demonstrate that words and concepts are not used in accordance with essentialism. The results are, however, consistent with "representational change" views of concepts, which are broadly Fregean in their motivation. We conclude that since essentialism is not vindicated by ordinary word use, it fails to undermine the cognitive psychology of concepts. PMID- 8706379 TI - Analysis of perceptual confusions between nine sets of consonant-vowel sounds in normal and dyslexic adults. AB - It is widely accepted that most developmental dyslexics perform poorly on tasks which assess phonological awareness. One reason for this association might be that the early or "input" phonological representations of speech sounds are distorted or noisy in some way. We have attempted to test this hypothesis directly. In Experiment 1, we measured the confusions that adult dyslexics and controls made when they listened to nine randomly presented consonant-vowel (CV) segments [sequence: see text] under four conditions of increasing white noise masking. Subjects could replay stimuli and were under no obligation to respond quickly. Responses were selected with a computer mouse from a set of nine letter strings, corresponding to the auditory stimuli, presented on a VDU. While the overall pattern of confusions made by dyslexics and controls was very similar for this stimulus set, dyslexics confused [sequence: see text] significantly more than did controls. In Experiment 2, subjects heard each stimulus once only and were forced to respond as quickly as possible. Under these timed conditions, the pattern of confusions made by dyslexics and controls was the same as before, but dyslexics took longer to respond than controls. The slower responses of dyslexics in Experiment 2 could have arisen because: (a) they were slower at processing the auditory stimuli than controls, (b) they had worse visual pattern memory for letter strings than controls, (c) they were slower than controls at using the computer mouse. In Experiments 3, 4 and 5 subjects carried out control tasks which eliminated each of these possibilities and confirmed that the results from the auditory tasks genuinely reflected subjects' speech perception. We propose that the fine structure of dyslexics' input phonological representations should be further explored with this confusion paradigm by using other speech sounds containing VCs, CCVs and VCCs. PMID- 8706380 TI - Drinking and driving don't mix: inductive generalization in infancy. AB - The traditional view of inductive generalization in infancy is that it rests on perceptual similarity; infants are said to form perceptually based categories, such as dogs and cats, and then to associate various properties with them. Superordinate-level inductions, such as generalizations about animals as a domain, have been considered to be more abstract and assumed to be a later achievement. Three experiments were conducted to investigate these issues, using 14-month-olds as subjects. We modeled various properties or actions appropriate to animals or to vehicles and then assessed whether infants were willing to generalize their imitations of these actions to different exemplars from the same and different domains. Contrary to the traditional view, we found that infants this age have generalized the properties of drinking and sleeping throughout the animal domain, and the properties of "being keyed" and "giving a ride" throughout the vehicle domain. These generalizations are constrained solely by the boundaries of the domains themselves and are not influenced by the perceptual similarity of exemplars within the domains. PMID- 8706381 TI - Where to look first for suggestibility in young children. AB - Preschoolers' suggestibility following exposure to biased information has often been interpreted as indicating that memory traces have been genuinely altered. However, young children may not recognize that the purpose and relevance of questions in experiments on suggestibility is to determine whether they can ignore misinformation in remembering the original details of stories. Instead, children may be prompted to regard the original story details as trivial by experimenters who are perceived as having portrayed these details as unimportant or irrelevant in that they themselves did not bother to get these right. Under such conditions, children may interpret the biased information to mean that a biased alternative was an acceptable, or even a preferred, test choice when compared to the original details. We report the results of an investigation with 3- to 5-year-olds in which children heard a story followed the next day by either biased, unbiased, or no information. The children were able to identify the original story details 6 days later when the questions were phrased in an explicit manner that referred to the time of the information to be recalled. PMID- 8706382 TI - Selected topics in dermatology for the primary care physician. PMID- 8706383 TI - New concepts in atopic dermatitis. PMID- 8706384 TI - Anticonvulsant hypersensitivity syndrome. PMID- 8706386 TI - The myriad faces of psoriatic arthritis. PMID- 8706385 TI - Diagnosis and management of dermatomyositis. PMID- 8706387 TI - Evaluation of cutaneous lymphomas. PMID- 8706388 TI - Porphyria cutanea tarda: a clinical review. PMID- 8706389 TI - Nonmelanoma skin cancer. PMID- 8706390 TI - Clinical management of herpes zoster in the elderly patient. PMID- 8706391 TI - Allergic contact cheilitis. AB - Allergic contact cheilitis describes a not uncommon inflammatory reaction of the lips. We reviewed its allergic causes, and suspect that the literature does not adequately document the frequency and etiologic factors involved. PMID- 8706392 TI - Baseline transepidermal water loss in patients with acute and healed irritant contact dermatitis. AB - To examine the skin barrier function of patients with acute and healed irritant contact dermatitis (n = 80) baseline transepidermal water loss (TEWL) was quantitatively measured using an evaporimeter. Healthy subjects served as controls (n = 40). Test areas were the forearm and the thigh. A significant increase in TEWL was observed in the patients with acute and with healed irritant contact dermatitis (ICD) as compared to healthy volunteers (P < or = 0.01). TEWL values in both test areas were comparable and markedly correlated (P < or = 0.01) with each other in every group. Thus, it is possible that basal TEWL depends more on the intrinsic skin barrier function of the subjects rather than the 2 anatomical regions examined. TEWL at the forearm with acute ICD was significantly higher (P < or = 0.01) than that of the group with healed ICD, but not for TEWL at the thigh suggesting that ICD may aggravate the barrier function of the adjacent uninvolved skin. It is assumed, that increased basal TEWL in patients with ICD may reflect a constitutional deviation of epidermal barrier function. This event seems to be comparable with the well-known symptom of atopic individuals. Using a detailed atopic scoring system in such a study may clarify the question of whether a proportion of patients with hand ICD may indeed be atopic individuals. PMID- 8706393 TI - Evaluation of patch test results by use of the reaction index. An analysis of data recorded by the Information Network of Departments of Dermatology (IVDK). AB - Results of patch tests performed in 15,553 patients by 18 dermatological departments (members of the German Contact Dermatitis Research Group) and recorded by the Information Network of Departments of Dermatology (IVDK) were analysed by comparing recently defined reaction indices (RIs). The RIs studied were calculated from the numbers of allergic (a), questionable (q), and irritant (i) reactions (RI = (a-q-i/(a + q + i)), which were obtained using 13 European standard allergens. RIs were calculated for all patients and for separate subgroups defined by age, sex, history of atopic dermatitis, time of allergen exposure (1 versus 2 days), and time of patch test reading (2 or 3 days after allergen application). Higher RIs were consistently obtained when patch tests were applied for 1 day, as compared to 2 days. Readings at 3 days after allergen application resulted in higher RIs than readings after 2 days. In contrast, sex, age, and history of atopic dermatitis of patients were not found to have a consistent influence on the RIs. We suggest that reading after 3 days should be obligatory, and that allergen exposure for 1 day instead of 2 days might make patch test evaluation easier. These suggestions need to be substantiated by data on clinical relevance. PMID- 8706394 TI - Adverse contact reactions to sculptured acrylic nails: 4 case reports and a literature review. AB - 4 cases with differing presentations of contact allergy to acrylates in sculptured acrylic nails are presented. These reactions include nail fold, fingertip and hand dermatitis, face and neck dermatitis, dystrophic nail changes and paraesthesia. We discuss acrylic nails and review the previously published reactions to acrylates in acrylic nails. PMID- 8706395 TI - The development and course of test reactions to gold sodium thiosulfate. AB - In our department, gold sodium thiosulfate has become the 2nd most common allergen in routinely patch tested dermatitis patients, with a rate around 10%. Test reactions to this compound often appear late, sometimes so late that active sensitization may be suspected. This study was performed to study the time course of the allergic reaction to gold sodium thiosulfate and to elucidate whether late test reactions mean active sensitization. 10 patients with contact allergy to gold sodium thiosulfate (0.5% pet.) were retested epicutaneously (e.c.) and intracutaneously (i.c.) with dilution series. The clinical course was followed for 2 months with initially short intervals, later more extended. During the entire study, 26 positive e.c. reactions were diagnosed. Within the 1st week, 17 (65%) were recorded. 12 reactions (46% of 26) were noted at the ordinary reading, 3 days after test application. After 10 days, another 9 reactions (35%) appeared. The patients with the latter reactions also had positive test reactions within the 1st week. After 2 months, 9 reactions remained. Out of 30 i.c. tests applied, 25 became positive within 1 week. 19 (76%) of these reactions changed in morphology from thin infiltrates to deep nodules. Another 4 nodules appeared in patients with previous negative i.c. tests. All 23 nodules remained after 2 months. E.c. and i.c. test reactions to gold sodium thiosulfate are long-lasting. Positive patch test reactions emerging after 10 days do not automatically imply active sensitization. To diagnose contact allergy to gold sodium thiosulfate, the ordinary reading at day 3 is insufficient; even reading at 1 week is insufficient and must be supplemented by a reading at 3 weeks. All the i.c. test reactions, however, appeared within 1 week and, in several, a dermal nodule was formed. PMID- 8706396 TI - A clinical and patch test study of contact dermatitis from traditional Chinese medicinal materials. AB - Putative allergens in, clinical manifestations of, and patch tests with traditional Chinese medicinal materials (CMM) causing contact dermatitis (CD) were studied. 14 patients with CMM CD and 351 controls were patch tested with the standard series of allergens of Beijing Medical University and the suspected drug, as is. It was found that most CMM CD was caused by the CMM frequently used topically for analgesic and anti-inflammatory effect. All patients patch tested with the suspected drug gave positive results. Some patients also reacted to fragrance mix, colophony, rubber mix and ammoniated mercury. The positivity rates of fragrance mix and colophony in CMM CD patients were significantly higher than in controls (55.6% versus 16.5% for fragrance mix, chi 2 = 11.86, p < 0.01; and 55.6% versus 5.1% for colophony, chi 2 = 29.35, p < 0.01), while not those of rubber mix and ammoniated mercury. Results indicated that the topical analgesic and anti-inflammatory CMM, especially those containing fragrance, may cause contact sensitization. Clinical findings in CMM CD included cutaneous irritation, allergic contact dermatitis, immediate contact reactions and systemic contact dermatitis. CMM CD can be diagnosed by patch testing the putative drug, as is. PMID- 8706398 TI - Atopic dermatitis, nickel sensitivity and xerosis as risk factors for hand eczema in women. AB - Knowledge of the relationship between a history of atopic dermatitis and nickel sensitivity as risk factors for hand eczema is limited. Our objective was to study the relative importance of atopic manifestations and nickel sensitivity as risk factors for hand eczema in women. From women referred for patch testing with a dental series, 100 consecutive nickel-positive and 95 nickel-negative patients were studied. A history of atopic manifestations and a history of 5 symptoms of hand eczema, i.e., dry erythemas, maculopapules, vesicles, erosions and hyperkeratoses were recorded. A history of atopic dermatitis was found to increase the risk of the 5 symptoms of hand eczema and sick leave due to hand eczema highly significantly. Nickel sensitivity increased only the risk of vesicles and erosions. Xerosis increased the risk of dry erythemas and vesicles. We concluded that a history of atopic dermatitis was more important than nickel sensitivity as a risk factor for hand eczema in women. PMID- 8706397 TI - Rosin (colophony) and zinc oxide in adhesive bandages. An appropriate combination for rosin-sensitive patients? AB - Adhesives and bandages containing zinc oxide (ZnO) and rosin are often used for treatment of ulcers. The aim of this work was to study a possible inhibitory effect of ZnO on the elicitation of allergic contact reactions to rosin and to determine whether such an effect might be due to the formation of zinc resinates. Patch testing in rosin-sensitive patients was performed with mixes of ZnO and rosin. The eliciting capacities of one mix in which zinc resinates might be formed and another mix in which the formation of zinc resinates was prevented, were compared to the eliciting capacity of rosin alone. If a reduction of the allergic response was observed for the mix in which zinc resinates might form and if no reduction was observed for the mix in which formation of zinc resinates was prevented, this would support the hypothesis that the reduction in eliciting capacity was due to formation of zinc resinates. However, we could not see any difference in eliciting capacity when comparing the 2 mixes with a test preparation of rosin. Commercial adhesives (bandages) containing ZnO and rosin were patch tested in 7 rosin-sensitive patients. The concentration of abietic acid in the adhesives was determined with HPLC. Abietic acid was detected in all but one of those declared to contain rosin. The patients reacted to the adhesives in which abietic acid was detected. For rosin-sensitive persons, the addition of ZnO to rosin-containing adhesives cannot be regarded as an appropriate measure to inhibit the elicitation of allergic reactions. PMID- 8706399 TI - Sensitization of mice to paraphenylenediamine and structurally-related compounds: adjuvant effects of vitamin A supplementation. AB - Allergic contact dermatitis from moderate and weak contact sensitizers is generally studied with guinea pigs, since they are readily sensitized to contact allergens. Mice, by contrast, are poor responders to weak contact allergens. However, the variety of in vitro murine systems as well as murine specific reagents make mice the preferable species. With the use of vitamin A supplementation, 2 protocols were developed which sensitized CBA/J female mice to paraphenylenediamine. Mice were sensitized by 5 daily topical applications to shaven dorsal skin. Alternately, mice were sensitized by 2 intraperitoneal injections of antigen pulsed spleen cells. Sensitization to paraphenylenediamine was determined by ear swelling following topical application. Vitamin A supplementation was found to be essential for optimum response. Lymph node and spleen cells from sensitized mice were capable of proliferating to paraphenylenediamine in vitro. With the use of vitamin A supplementation and intraperitoneal injection, CBA/J mice were also sensitized to a number of compounds structurally related to paraphenylenediamine, including the ortho- and meta-derivatives of paraphenylenediamine, as well as hydroquinone and resorcinol. These new protocols, combined with vitamin A supplementation, expand the use of mice to study moderate sensitizers with minimal animal utilization. PMID- 8706401 TI - Contact allergy to cocamidopropyl betaine. AB - Cocamidopropyl betaine is an amphoteric surfactant used increasingly in cosmetic products. We describe 20 cases of cosmetic allergy to cocamidopropyl betaine; all were caused by shampoo or shower gel. 8 patients were hairdressers, who had occupational allergic contact dermatitis from shampoos. We recommend patch testing cocamidopropyl betaine 1% aq. routinely in hairdressers with dermatitis of the hands, and in all patients suspected of suffering from cosmetic allergy. PMID- 8706400 TI - Occupational hand dermatitis in a tertiary referral dermatology clinic in Taipei. AB - Occupational skin disease is one of the most common occupational diseases. The hand is the most frequent site of involvement in occupational skin disease. We interviewed and examined patients seen in the Contact Dermatitis Clinic of the National Taiwan University Medical Center, a tertiary referral center in Taipei City. For patients suspected of having allergic skin diseases, patch testing was carried out using the European standard series and suspected allergens. Occupational hand dermatitis (OHD) was diagnosed according to medical history, work exposure, physical examination, and patch test findings. 36% of patients seen were diagnosed as having OHD. Electronics, hairdressing, medical, chemical, and construction were the most important industries causing OHD. In the 164 patients with OHD, 58.5% had irritant contact dermatitis (ICD) and 41.5% allergic contact dermatitis (ACD). Dorsal fingers, nail folds, and dorsal hands were most frequently involved in patients with ACD; dorsal fingers, volar fingers and fingertips were most frequently involved in those with ICD. Using logistic regression analysis, we were able to identify the most important clinical presentations that predicted the types of OHD, ACD versus ICD. Patients with atopic history and palm involvement were more likely to have ICD, and those with nail fold involvement more likely to have ACD. In patients with ACD, the most important allergens were dichromate, nickel, cobalt, fragrance mix, epoxy resin, thiuram mix, and p-phenylenediamine. In this study, we identified the important industries and causal agents for OHD. Future preventive measures focused on these industries and agents to reduce OHD will be warranted. PMID- 8706403 TI - Allergic contact dermatitis from iroko and pine wood dust. PMID- 8706402 TI - Oral lichenoid lesions caused by allergy to mercury in amalgam fillings. AB - Oral lichenoid lesions (OLL) or lichen-planus-like lesions are often idiopathic. Our aim was to determine whether OLL can be caused by allergy to mercury in amalgam fillings, and whether resolution of OLL occurs after replacement of amalgam with other dental fillings. Patients with only OLL (except for 1 case with cutaneous lichen planus) referred for patch testing during 1985-1994 to the Contact and Occupational Dermatitis Clinic of the Skin & Cancer Foundation, Darlinghurst, were reviewed. Patch tests were performed with 1% mercury, 1% ammoniated mercury, 0.1% thimerosal, 0.1% mercuric chloride, 0.05% phenylmercuric nitrate and an amalgam disc, using Finn Chambers occluded for 2 days, 19 patients (17 women and 2 men; age range: 28-72 years) had OLL in close contact with amalgam fillings and showed positive patch test reactions to mercury compounds, 16 out of 19 patients had their amalgam fillings replaced. In 13 patients, the OLL healed. 1 patient had marked improvement. 1 patient had no improvement and developed multiple oral squamous cell carcinoma. In conclusion, OLL can be caused by allergy to mercury in amalgam fillings. Replacement of amalgam with other dental fillings usually results in resolution of OLL and is recommended for cases with positive patch test reactions to mercury compounds. PMID- 8706404 TI - Probable active sensitization to tixocortol pivalate. PMID- 8706405 TI - Contact sensitivity to food additives can cause oral and perioral symptoms. PMID- 8706406 TI - Generalized contact dermatitis from acetarsone. PMID- 8706407 TI - Generalized dermatitis due to mitomycin C patch test. PMID- 8706408 TI - Airborne erythema-multiforme-like eruption due to pyrethrum. PMID- 8706409 TI - HLA DR, DQA, DQB and DP antigens in patients allergic to nickel. PMID- 8706410 TI - Allergic contact dermatitis from butoxyethyl nicotinic acid and Centella asiatica extract. PMID- 8706411 TI - Contact urticaria from beer. PMID- 8706412 TI - Granulomatous dermatitis after implantation of a titanium-containing pacemaker. PMID- 8706413 TI - Persistent allergic contact dermatitis at the site of primary sensitization with squaric acid dibutyl ester. PMID- 8706414 TI - Occupational allergic contact dermatitis from vitamin A acetate. PMID- 8706415 TI - Allergic contact dermatitis due to thymol. PMID- 8706416 TI - Allergic contact dermatitis from a beeswax nipple-protective. PMID- 8706417 TI - Allergic contact dermatitis from tacalcitol. PMID- 8706418 TI - Allergic contact dermatitis and systemic contact dermatitis from cinchocaine. PMID- 8706419 TI - Adding p-aminoazobenzene may increase the sensitivity of the European standard series in detecting contact allergy to dyes, but carries the risk of active sensitization. PMID- 8706420 TI - Occupational allergic contact urticaria caused by cockroach (Blaberus giganteus). PMID- 8706421 TI - Exfoliative dermatitis caused by carbamazepine in a patient with isolated IgA deficiency. PMID- 8706422 TI - Patch testing in an unusual case of toxic epidermal necrolysis. PMID- 8706423 TI - Erythema-multiforme-like eruption following allergic contact dermatitis from sesquiterpene lactones in herbal medicine. PMID- 8706424 TI - Contact dermatitis due to 3-amino-methyl-3,5,5-trimethylcyclohexylamine (isophoronediamine; IPD) PMID- 8706425 TI - Maternal serum screening for birth defects: results of a Connecticut regional program. AB - Second trimester maternal serum screening provides a method to identify pregnancies at high risk for fetal Down's syndrome, trisomy 18, open neural tube defects, and a variety of other chromosomal and nonchromosomal fetal anomalies. Results are presented for a regional program to identify high-risk pregnancies using alpha feto-protein (AFP), human chorionic gonadotropin (hCG), and unconjugated estriol (uE3) analyses (triple marker testing). A total of 27,140 women received screening. Using a midtrimester Down's syndrome risk of 1:270 to define the high-risk group, 5.26% of women of all ages were screen-positive for Down's syndrome resulting in the eventual detection of approximately 72% of the affected fetuses. The detection rate for patients under 35 at estimated date of delivery was 61% and for women 35, or older, the detection rate was 100%. A separate protocol to screen for trisomy 18 identified 0.2% of pregnancies, with 38% of the trisomy 18 cases present in this group. Over 3% of women screen positive for Down's syndrome or trisomy 18 had a serious fetal chromosome anomaly. In addition, 2.89% of women had an elevated AFP (greater or equal to 2.0 multiples of median). This component of the screening resulted in the identification of 86% of the neural tube defects, 75% of the ventral wall defects, and also some of the other various fetal anomalies present in the screened population. Since both laboratory and clinical data are combined to generate patient-specific risks, there is a need for quality control elements that go beyond that normally required for a clinical laboratory alone. We stress the need for comprehensive follow-up programs to evaluate screening programs and maintain high quality. PMID- 8706426 TI - BCG against tuberculosis: 1996. PMID- 8706427 TI - Long-term complication following subtotal pancreatectomy for nesidioblastosis: a case report. AB - Nesidioblastosis is the most common cause of neonatal hypoglycemia. Although medical therapy has been attempted, it is generally accepted that these infants should undergo a subtotal to near total pancreatectomy with splenic preservation. Complications from this procedure have been few, most commonly those associated with decreased insulin production (diabetes). We describe a case of a young, white male who presented with complaints of hematemesis and melena 18 years following subtotal pancreatectomy for nesidioblastosis. PMID- 8706428 TI - Loop diuretics: comparison of torsemide, furosemide, and bumetanide. PMID- 8706429 TI - Tips and tricks: the hard way--ABCs of quadriplegia care. AB - The following are things I have learned, routines my nurses have brought to me, or that we have developed together during 11-years existence with amyotropic lateral sclerosis (ALS) eight of which have been burdened by ALS induced quadriplegia and ventilator dependence. Items are listed, not in order of importance, which may vary from individual to individual, but alphabetically. Some are unique. Some commonplace. All are useful and many are of surprising value. Deviations from common understanding or practice are marked with an asterisk, and practices that can afford significant economies are double asterisked. This is written in an attempt to share with others in the profession lessons I have learned from personal experience--the hard way. PMID- 8706430 TI - The new medicine. PMID- 8706431 TI - "Do not think, but try; be patient, be accurate". PMID- 8706432 TI - "Philosophers will be busy". PMID- 8706433 TI - In response to "deconstructing the Hippocratic oath". PMID- 8706434 TI - Connecticut Supreme Court limits liability of mental health care givers. PMID- 8706435 TI - Cyclic progestin administration brings about luteinization during continuous antiprogestin treatment. AB - The antiprogestin RU 486 has been proposed for use as a contraceptive because it disturbs folliculogenesis and inhibits ovulation when given continuously. In order to achieve regular and predictable withdrawal bleeding and to counteract the continuous influence of estrogen on the endometrium, a sequential, cyclic progestin regimen has been added. The drawback has been the frequent rise in the concentration of serum P, suggesting ovulation. The present pilot study with three subjects was carried out to test whether simultaneous administration of RU 486 and progestin during the second half of the cycle would eliminate the rise in serum P concentrations. Based on elevated serum P values, five of the seven cycles were apparently ovulatory. RU 486 alone, when given throughout the cycle at a dose of 8 mg per day without added progestin, resulted in anovulatory cycles, but the lengths of these cycles were prolonged. We conclude that both simultaneous and sequential cyclic co-administration of progestin with RU 486 abolishes the antiovulatory action of RU 486. This decreases the possibility of using antiprogestins in non-estrogenic contraceptive regimens (unless contraceptive effects of RU 486 exist at the endometrial level). PMID- 8706436 TI - Performance of the TCu380A and Cu-Fix IUDs in an international randomized trail. AB - To compare the clinical performance of the TCu380A, the most widely used copper IUD in the world today, and a new frameless device, the Cu-Fix, we report results from a randomized international two-year clinical trial involving 874 parous women followed for an average of fourteen months. Event rates at the end of two years (per 100 women) for pregnancy were 0.0 for the TCu380A and 1.5 for the Cu Fix. Termination due to expulsion was significantly less for TCu380A as compared with Cu-Fix users (2.0 and 11.4 per hundred women, respectively); other medical reasons for termination (bleeding/pain, pelvic inflammatory disease (PID), and other) did not differ significantly. PID incidence rates at two years were 1.0 per hundred women for the TCu380A and 1.6 for the Cu-Fix (equivalent to 6.0 and 8.3 per 1,000 woman-years, respectively). The net cumulative continuation rates at two years per hundred women were 78.8 for TCu380A, and 71.9 for the Cu-Fix. Both IUDs provide highly effective protection against pregnancy, but the TCu380A has a lower expulsion rate. The low rate of PID indicates that it is an in frequent occurrence in appropriately selected candidates. PMID- 8706437 TI - Spermicidal activity of Magainins: in vitro and in vivo studies. AB - Spermicidal efficacy of two synthetic antimicrobial peptides, viz. Magainin-A (Mag-A) and Magainin-G (Mag-G), have been studied under in vitro condition using rat and human spermatozoa. The data showed that sperm motility was inhibited with various concentrations of Magainins at different intervals ranging from 20 to 480 seconds as compared to controls. The minimum concentration of Mag-A required to immobilize spermatozoa (40-60 x 10(6)/ml of saline) within 20 seconds was found to be 100 micrograms in rat and 200 micrograms in human. The action of Mags on sperm motility was observed to be dose-dependent. The activity of Mag-A was significantly (P < 0.05) higher than Mag-G (P < 0.05). The in vivo studies in rats indicated that Mag-A when applied intravaginally, on the day of estrus, at the dose level of 200 micrograms, for five consecutive estrous cycles, induced no abnormalities either in the length of estrous cycle or in morphology of vaginal epithelial cells. When Mag-A was applied intravaginally before mating on the proestrusestrus day, complete arrest of sperm motility with 200 micrograms concentration was observed and none of these treated females conceived after the mating. In view of these facts, it is suggested that Mag-A, having anti bacterial, anti-viral, anti-fungal and spermicidal activities, could be a potent vaginal contraceptive. PMID- 8706438 TI - Utilization patterns of oral contraceptives in UK general practice. AB - The aim of this article was to analyze and compare recent oral contraceptive utilization patterns in UK general practice. The database comprised at least 12 months of medical records during 1990-1991 from 693-705 women aged 14-45 years registered with 398 practices. The database was searched for OC utilization patterns with emphasis on differences between combined OCs containing levonorgestrel (LNG), desogestrel (DSG), or gestodene (GSD). Women using OCs with LNG were older, more likely to be long-term users, and less likely to be starters or switchers than women using OCs with DSG or GSD. Use patterns of triphasic LNG did not differ markedly from that of monophasic LNG-containing OCs. Use of Schering PC4 a postcoital preparation peaked among women < 20 years, whereas progestogen-only pills were prescribed mainly to women > 30 years. The different utilization patterns between women using LNG-containing OCs and those using OCs containing DSG or GSD should be taken into account when making comparisons of the incidences of side effects. PMID- 8706439 TI - Who gives advice about postpartum contraception? AB - Women were interviewed to determine what advice they received about postpartum contraception and what they thought of it. Only 4% of women discussed postpartum contraception antenatally. Up to 84% discussed the issue with a midwife on the postnatal ward but discussion was often felt to be brief, limited and frequently held as the mother was leaving the hospital. Obstetricians appeared to have little interest in the subject and only 50% of mothers left the hospital with supplies of a contraceptive. Almost all women discussed contraception with their general practitioner at the postnatal check but a significant number felt that the choice of method was limited to condoms or pills. The postnatal check is traditionally held at six weeks--two to three weeks after the recommended time for starting contraceptive precautions. Women with short inter-pregnancy intervals were younger, less likely to be married and more likely to default from postnatal follow-up. Pregnant women should be offered the opportunity during the antenatal period to discuss postpartum contraception with someone who has a special interest in the subject. The postnatal ward is not an appropriate setting for discussion about future contraception. PMID- 8706440 TI - Latex rubber condoms: predicting and extending shelf life. AB - Condoms from five manufacturers were subjected to controlled exposures of heat, humidity, and air and to different natural environments in five countries. Under aerobic conditions (condoms in permeable packages or unpackaged), stress properties declined. The relationship between rate of decline as a function of temperature was quadratic. Under oxygen-restricted conditions (foil-wrapped packages) at average storage temperatures of 30 degrees C and lower, strain properties declined with little or no significant change in stress properties. The effect is to cause condoms to become stiffer; high-breakage rates in use have been correlated with product stiffening. A new rationale for accelerated-aging tests to predict condom shelf stability is suggested, including a test to control the trend of condoms to stiffen. Silicone lubricant, impermeable packaging, and inclusion of antioxidants in the condom formulation can prevent or minimize aerobic breakdown of latex condoms. Specifying low-modulus condoms can prevent excessive stiffening. PMID- 8706441 TI - Zinc acetate and lyophilized aloe barbadensis as vaginal contraceptive. AB - Twenty samples of fresh ejaculate, donated by healthy volunteers ranging in age from 20-30 years, were obtained from the Center for Fertility & Cryobiology, University of Missouri, Columbia, Missouri. Average semen volume was 2.49 ml; average sperm motility was 71.32%; and average sperm density was 113.71 x 10(6) /ml. Testing for spermicidal effectiveness of a 1% concentration of zinc acetate, zinc sulfate, zinc chloride, and zinc gluconate proved that only zinc acetate was spermicidal. It appears this is due to the acetate in zinc acetate which may decrease oxygen utilization by sperm. Zinc acetate in vitro was antiviral while lyophilized aloe barbadensis was not. Lyophilized aloe barbadensis at concentrations of 7.5% and 10% proved to be spermicidal due to the multiple micro elements (boron, barium, calcium, chromium, copper, iron, potassium, magnesium, manganese, phosphorus, and zinc) which were toxic to the tail causing instant immobilization. The two compounds did not irritate or cause ulceration of rabbit vaginal epithelium. These results suggest the possibility of using zinc acetate and lyophilized aloe barbadensis as a new, effective and safe vaginal contraceptive. PMID- 8706442 TI - The use of misoprostol for termination of early pregnancy. AB - Misoprostol is a prostaglandin analogue which has uterotonic properties. Administered vaginally, it is an effective agent for induction of second trimester abortions. This study was undertaken to determine if the vaginal administration of misoprostol is effective for inducing first trimester abortions. Fifty-eight women with pregnancies less than 10 weeks gestation who desired pregnancy termination received varying dosages of vaginal misoprostol, either alone or in combination with laminaria or tamoxifen. The overall success rate for a complete abortion was 61%. The use of laminaria or tamoxifen did not affect success rates. Abortions occurred within 24 hours of administration of misoprostol. Side effects were minimal. There were no significant differences in any of the following between those who had a successful abortion and those who did not: gravidity, parity, prior elective abortion, age, gestational age of the pregnancy, and level of human chorionic gonadotropin. Although not as successful as other combination regimens, misoprostol alone is readily available, easy to administer, and without major side effects. The use of this agent could eliminate the need for about two-thirds of surgical abortions in the first 10 weeks of gestation. PMID- 8706443 TI - Evaluation of the effectiveness of vaginal misoprostol to induce first trimester abortion. AB - Two doses, 200 and 400 micrograms, of misoprostol, administered vaginally every 12 hours, up to four times, were tested in 101 and 133 healthy women, respectively, for interruption of pregnancies with 35 through 77 days of amenorrhea. The proportion of women who aborted increased with longer duration of treatment and was significantly higher with 400 than with 200 micrograms (66 versus 46 percent at 48 hours). Significance was maintained after controlling by age, body weight, parity, previous abortion and gestational age. Abortions were classified as incomplete or complete, according to the presence or not of embryonic tissue in the uterine cavity, diagnosed by vaginal sonography. Vacuum aspiration was carried out in all cases not classified as complete abortion 48 hours after the initiation of treatment, or earlier in case of persistent bleeding or woman's request. The possibility of increasing effectiveness by using higher dose, shorter intervals or longer duration of treatment is discussed. PMID- 8706444 TI - Antifertility effects of (+)-S-2-amino-6-iodoacetamidohexanoic acid (2-AIHA) in female rats. AB - (+)-S-2-amino-6-iodoacetamidohexanoic acid (AIHA), an irreversible inhibitor of the ornithindecarboxylase and extrahepatic arginase enzymatic activities with antineoplasic properties, was evaluated for antifertility activity in pregnant rats by oral administration at different periods of gestation. Our results showed that doses of 10 and 20 mg/kg of AIHA orally administered produced a contraceptive effect when it was administered from days 2 to 5, and 8 to 12 of gestation, respectively. The gestation time was slightly shortened when AIHA was applied from day 15 until labor. No sign of external malformations in fetuses was observed. On the other hand, AIHA did not affect the total length of oestrous cycle at the same dosage level used to interrupt pregnancy. In ovariectomized immature rats, neither changes in uterine weight, premature vaginal opening, or cornified cells were found. However, AIHA enhanced the estradiol-induced increase in uterine weights when both were concomitantly administered. PMID- 8706445 TI - Post hoc analyses in sepsis trials: a formula for disappointment? PMID- 8706446 TI - Judicial intervention in medical decision-making: a failure of the medical system? PMID- 8706447 TI - Assessment of the safety and efficacy of the monoclonal anti-tumor necrosis factor antibody-fragment, MAK 195F, in patients with sepsis and septic shock: a multicenter, randomized, placebo-controlled, dose-ranging study. AB - OBJECTIVE: To investigate the safety, biological effects, and efficacy of the anti-tumor necrosis factor (TNF) antibody fragment, MAK 195F, in a phase II trial in patient with severe sepsis. DESIGN: Prospective, randomized, open label, placebo-controlled, dose-ranging, multicenter, multinational clinical trial. SETTING: Sixteen academic medical centers' intensive care units in six European countries. PATIENTS: One hundred twenty-two patients with severe sepsis or septic shock who received standard supportive care and antimicrobial therapy. INTERVENTIONS: Patients received one of three different doses of the anti-TNF antibody (0.1 mg/kg, 0.3 mg/kg, or 1.0 mg/kg) or placebo; the antibody or placebo was given in nine doses at 8-hr intervals over 3 days. MEASUREMENTS AND MAIN RESULTS: There were no significant differences in mortality rates among the groups receiving various doses of the anti-TNF antibody or placebo, but patients with baseline serum interleukin (IL)-6 concentrations of > 1000 pg/mL appeared to benefit from MAK 195F in a dose-dependent fashion. Increased circulating IL-6 concentrations, but not TNF concentrations, were found to be important prognostic indicators for mortality for the patients in the placebo and the two lower dosage groups but not in the high dosage group (1 mg/kg). IL-6 concentrations decreased during the first 24 hrs of treatment in all three anti-TNF groups but not in the placebo group. MAK 195F was well tolerated by all patients. Human antimurine antibodies developed in 40% of the patients receiving the antibody. CONCLUSIONS: There was no increase in survival from sepsis for the patients receiving anti-TNF treatment in the overall study population. Retrospective stratification of patients by IL-6 concentrations suggests beneficial effects of the drug for patients with baseline circulating IL-6 concentrations of > 1000 pg/mL. This hypothesis requires validation in a larger, blinded, prospective study. PMID- 8706448 TI - PRISM III: an updated Pediatric Risk of Mortality score. AB - OBJECTIVES: The relationship between physiologic status and mortality risk should be reevaluated as new treatment protocols, therapeutic interventions, and monitoring strategies are introduced and as patient populations change. We developed and validated a third-generation pediatric physiology-based score for mortality risk, Pediatric Risk of Mortality III (PRISM III). DESIGN: Prospective cohort. SETTING: There were 32 pediatric intensive care units (ICUs): 16 pediatric ICUs were randomly chosen and 16 volunteered. PATIENTS: Consecutive admissions at each site were included until at least 11 deaths per site occurred. MEASUREMENTS AND MAIN RESULTS: Physiologic data included the most abnormal values from the first 12 and the second 12 hrs of ICU stay. Outcomes and descriptive data were also collected. Physiologic variables where normal values change with age were stratified by age (neonate, infant, child, adolescent). The database was randomly split into development (90%) and validation (10%) sets. Variables and their ranges were chosen by computing the risk of death (odds ratios) relative to the midrange of survivors for each physiologic variable. Univariate and multivariate statistical procedures, including multiple logistic regression analysis, were used to develop the PRISM III score and mortality risk predictors. Data were collected on 11,165 admissions (543 deaths). The PRISM III score has 17 physiologic variables subdivided into 26 ranges. The variables most predictive of mortality were minimum systolic blood pressure, abnormal pupillary reflexes, and stupor/coma. Other risk factors, including two acute and two chronic diagnoses, and four additional risk factors, were used in the final predictors. The PRISM III score and the additional risk factors were applied to the first 12 hrs of stay (PRISM III-12) and the first 24 hours of stay (PRISM III-24). The Hosmer Lemeshow chi-square goodness-of-fit evaluations demonstrated absence of significant calibration errors (p values: PRISM III-12 development = .2496; PRISM III-24 development = .1374; PRISM III-12 validation = .4168; PRISM III-24 validation = .5504). The area under the receiver operating curve and Flora's z statistic indicated excellent discrimination and accuracy (area under the receiver operating curve - PRISM III-12 development 947 +/- 0.007; PRISM III-24 development 0.958 +/- 0.006; PRISM III-12 validation 0.941 +/- 0.021; PRISM III 24 validation 0.944 +/- 0.021; Flora's z-statistic - PRISM III-12 validation = .7479; PRISM III-24 validation = .9225), although generally, the PRISM III-24 performed better than the PRISM III-12 models. Excellent goodness-of-fit was also found for patient groups stratified by age (significance levels: PRISM III-12 = .1622; PRISM III-24 = .4137), and by diagnosis (significance levels: PRISM III-12 = .5992; PRISM III-24 = .7939). CONCLUSIONS: PRISM III resulted in several improvements over the original PRISM. Reassessment of physiologic variables and their ranges, better age adjustment for selected variables, and additional risk factors resulted in a mortality risk model that is more accurate and discriminates better. The large number of diverse ICUs in the database indicates PRISM III is more likely to be representative of United States units. PMID- 8706449 TI - Does selective decontamination of the digestive tract reduce mortality for severely ill patients? AB - OBJECTIVE: To investigate the relationship between baseline risk of death and reduced mortality after selective decontamination of the digestive tract in intensive care unit patients. DESIGN: Analysis of data from a meta-analysis of 23 randomized, controlled trials. PATIENTS: A total of 4,142 adult intensive care unit patients from the 23 trials. MEASUREMENTS AND MAIN RESULTS: Mortality for patients receiving selective decontamination of the digestive tract treatment was analyzed as a function of baseline risk of death at study entry, using weighted least squares regression across all 23 trials. In testing whether the slope of the regression is different than 1.0, the observed t value is 3.32 (p < .004), suggesting that the efficacy of selective decontamination of the digestive tract in reducing mortality is significantly better in populations at high mortality risk at study entry. CONCLUSIONS: Mortality reduction from selective decontamination of the digestive tract appears related to the mortality risk of patients at the time of study entry. Future trials should consider using baseline risk assessment as part of trial design and outcome analysis. PMID- 8706450 TI - Physiology and pharmacokinetics of a novel hemoglobin-based oxygen carrier in humans. AB - OBJECTIVE: To evaluate the physiology and pharmacokinetics of a novel hemoglobin based oxygen carrier of bovine origin. DESIGN: Randomized, single-blind, placebo controlled, dose-escalation study. SETTING: The Upjohn Research Clinics (Kalamazoo, MI). SUBJECTS: Normal healthy adult men between the ages of 18 and 45 yrs. There were 18 subjects who received active treatment and 23 controls. INTERVENTIONS: All subjects had phlebotomy of 15% of blood volume (performed in <15 mins) followed by isovolemic hemodilution (3:1, Ringer's lactate to the volume of whole blood removed) over a 90-min period, and either active drug (polymerized bovine hemoglobin) or a control infusion of lactated Ringer's solution (each infusion given over a total of 4.3 hrs). The subjects randomized to active treatment received a loading dose and a continuous infusion of polymerized bovine hemoglobin for a total dose of 16.5, 24.1, 30.2, 38.0, or 45.0 g. All subjects had an indwelling radial artery catheter (for blood pressure and arterial blood gas measurements), determination of cardiac function (by impedance plethysmography), serial pulmonary function tests (spirometry and diffusion capacity), and metabolic cart measurements. MEASUREMENTS AND MAIN RESULTS: Pharmacokinetics of the plasma bovine hemoglobin demonstrated that the elimination of the hemoglobin-based oxygen carrier was a linear, first-order process and that there was no renal excretion. Peak plasma concentrations were between 1 to 2 g/dL and plasma half-life approached 20 hrs at the highest doses given. Diffusion capacity of oxygen was increased up to 20% above baseline in the 38.0 and 45.0 g groups in comparison with controls (approximately 14% below baseline) between 2 and 24 hrs after the infusion (p < .01). Other pulmonary function tests and arterial blood gas measurements were unremarkable. Arterial oxygen content and oxygen delivery tended to be greater in active groups than in controls. CONCLUSIONS: The plasma concentrations of bovine hemoglobin were directly proportional to the doses administered. An increase in diffusion capacity paralleled the plasma bovine hemoglobin concentrations. Dosing of the hemoglobin-based oxygen carrier of bovine origin to a target plasma hemoglobin concentration can be achieved using pharmacokinetic principles with measurable effects on oxygen physiology. PMID- 8706451 TI - High plasma tumor necrosis factor (TNF)-alpha concentrations and a sepsis-like syndrome in patients undergoing hyperthermic isolated limb perfusion with recombinant TNF-alpha, interferon-gamma, and melphalan. AB - OBJECTIVES: To describe the postoperative course of patients who underwent hyperthermic isolated limb perfusion with recombinant tumor necrosis factor (TNF) alpha and melphalan after pretreatment with recombinant interferon-gamma as treatment for recurrent melanoma, primary nonresectable soft-tissue tumors, planocellular carcinoma, or metastatic carcinoma. To measure systemic TNF-alpha concentrations and relate these values with indices of disease severity. SETTING: A 12-bed surgical intensive care unit (ICU) in a university referral hospital. DESIGN: Prospective, descriptive study. PATIENTS: Consecutive patients (n=25) treated with hyperthermic isolated limb perfusion. INTERVENTIONS: Blood samples were taken at regular intervals to determine TNF-alpha concentrations during and after hyperthermic isolated limb perfusion with recombinant TNF-alpha. Hemodynamic variables were obtained with a Swan-Ganz pulmonary artery catheter. MEASUREMENTS AND MAIN RESULTS: All patients developed features of sepsis syndrome and required intensive care treatment. Most patients recovered quickly, with a median ICU stay of 2 days (range 1 to 25). Maximum systemic TNF-alpha concentrations ranged from 2284 to 83,000 ng/L (median 25,409) and returned to baseline values within 8 hrs. Despite these high concentrations of TNF-alpha, no patient died in the ICU, although the patient with the highest TNF-alpha concentration developed multiple organ failure and required continuous venovenous hemofiltration for 16 days. Linear regression analysis showed positive correlations between maximum TNF-alpha concentrations and systemic vascular resistance (p < .01), cardiac index (p < .02), Lung Injury Score (p < .02), prothrombin time (p < .02), and activated partial thromboplastin time (p < .05). CONCLUSIONS: Hyperthermic isolated limb perfusion with recombinant TNF-alpha leads to high systemic concentrations of TNF-alpha, probably due to leakage of recombinant TNF-alpha from the perfusion circuit, mainly through collateral blood flow. A sepsis-like syndrome is seen in all patients. Despite high concentrations of systemic TNF-alpha, this sepsis syndrome is short-lived and recovery is rapid and complete in most patients. PMID- 8706452 TI - Closed-loop control of airway occlusion pressure at 0.1 second (P0.1) applied to pressure-support ventilation: algorithm and application in intubated patients. AB - OBJECTIVE: Airway occlusion pressure at 0.1 sec (P0.1) is an index of respiratory center output. During pressure-support ventilation, P0.1 correlates with the mechanical output of the inspiratory muscles and has an inverse relationship with the amount of pressure-support ventilation. Based on these observations, we designed a closed-loop control which, by automatically adjusting pressure-support ventilation, stabilizes P0.1, and hence patient inspiratory activity, at a desired target. The purpose of the study was to demonstrate the feasibility of the method, rather than its efficacy or even its influence on patient outcome. DESIGN: Prospective, randomized trial. SETTING: A general intensive care unit of a university hospital in Italy. PATIENTS: Eight stable patients intubated and ventilated with pressure-support ventilation for acute respiratory failure. INTERVENTIONS: Patients were transiently connected to a computer-controlled ventilator on which the algorithm for closed-loop control was implemented. The closed-loop control was based on breath by breath measurement of P0.1, and on comparison with a target set by the user. When actual P0.1 proved to be higher than the target value, the P0.1 controller automatically increased pressure support ventilation, and decreased it when P0.1 proved to be lower than the target value. For safety, a volume controller was also implemented. Four P0.1 targets (1.5, 2.5, 3.5, and 4.5 cm H2O) were applied at random for 15 mins each. MEASUREMENTS AND MAIN RESULTS: The closed-loop algorithm was able to control P0.1, with a difference from the set targets of 0.59 +/- 0.27 (SD) cm H2O. CONCLUSIONS: The study shows that P0.1 can be automatically controlled by pressure-support ventilation adjustments with a computer. Inspiratory activity can thus be stabilized at a level prescribed by the physician. PMID- 8706453 TI - Volume-controlled versus biphasic positive airway pressure ventilation in leukopenic patients with severe respiratory failure. AB - OBJECTIVE: To study comparatively the effects of volume-controlled vs. biphasic positive airway pressure mechanical ventilation on respiratory mechanics and oxygenation in leukopenic patients with severe respiratory failure. DESIGN: Prospective, comparative study. SETTING: Medical intensive care unit of a university hospital. PATIENTS: Leukopenic (<1000 leukocytes/microliter) patients (n=20) after cytoreductive chemotherapy requiring mechanical ventilation for severe respiratory failure (Murray score of > 2.5). INTERVENTION: Patients were assigned in a consecutive, alternating manner to receive either volume-controlled or biphasic positive airway pressure mechanical ventilation, starting within 12 to 24 hrs after endotracheal intubation. MEASUREMENTS AND MAIN RESULTS: Tidal volume, inspiratory flow, peak inspiratory and positive end-expiratory pressures, FIO2, and arterial blood gas analyses were recorded hourly for a study period of 48 hrs. Biphasic positive airway pressure ventilation was associated with a significant reduction in peak inspiratory pressure (mean differences at 24, 36, and 48 hrs: 4.4, 3.4, and 4.2 cm H2O; p = .024, .019, and .013, respectively) and positive end-expiratory pressures (mean differences at 24, 36, and 48 hrs: 1.6, 1.4, and 1.5 cm H20; p = .023, .024, and .023, respectively) at significantly lower FIO2 (mean differences at 12, 24, 36, and 48 hrs; p = .007, .015, .016, and .011, respectively). PaO2/FIO2 ratios and CO2 removal were similar under ventilatory conditions. CONCLUSIONS: Biphasic positive airway pressure ventilation offers the advantage of significantly reduced peak inspiratory and positive end-expiratory pressures at a lower FIO2 and with at least similar oxygenation and CO2 removal as achieved by volume-controlled mechanical ventilation. Our results are in line with previous reports on nonleukopenic patients and suggest that the positive effects of pressure-limited mechanical ventilation are independent of circulating white blood cells. Further studies are mandatory to demonstrate clinical benefit in this critically ill patient population. PMID- 8706454 TI - Relationship between arterial carbon dioxide and end-tidal carbon dioxide in mechanically ventilated adults with severe head trauma. AB - OBJECTIVE: To examine the agreement and association of a noninvasive method of measuring CO2 (using end-tidal PCO2) with PaCO2 in mechanically ventilated adults with severe head trauma. DESIGN: A prospective, quasi-experimental, repeated measures study was used to compare end-tidal PCO2 and PaCO2 at two time points: before and after a standardized endotracheal suctioning procedure. INTERVENTIONS: Controlled intervention of endotracheal suctioning. SETTING: The study was conducted at two intensive care units designated as Level 1 trauma centers. PATIENTS: A consecutive sample of 35 severe head-injured patients with a Glasgow Coma Scale score of < or = 8. MEASUREMENTS AND MAIN RESULTS: End-tidal PCO2 and PaC02 values were simultaneously obtained and compared. End-tidal PCO2 was measured, using a sidestream sensor placed in line of the ventilator circuit's deadspace. Arterial gases were drawn from an indwelling arterial catheter. No relationship was found between arterial and end-tidal measures (range r2 = .09 to r2 = .11). Using the Bland-Altman technique, agreement decreased as the amount of positive end-expiratory pressure increased. When a subset of patients (mechanically ventilated, with positive end-expiratory pressures of < 5 cm H2O, paralyzed, and sedated) were examined (n = 12), the correlation between the CO2 measures improved (r2 = .77). CONCLUSIONS: This study indicated that end-tidal PCO2 monitoring correlated well with PaCO2 in patients without respiratory complications or without spontaneous breathing, resulting in rebreathing of gases. However, its clinical validity is questionable in patients who have the greatest need for end-tidal PCO2 monitoring (i.e., patients who have respiratory distress or who are breathing spontaneously and overriding the ventilator. PMID- 8706455 TI - End-tidal carbon dioxide during cardiopulmonary resuscitation in humans presenting mostly with asystole: a predictor of outcome. AB - OBJECTIVE: To determine whether continuous semiquantitative assessment of end tidal CO2 could provide a highly sensitive predictor of return of spontaneous circulation during cardiopulmonary resuscitation (CPR). DESIGN: Prospective, clinical study. SETTING: Prehospital CPR. PATIENTS: One hundred twenty patients, during nontraumatic cardiac arrest. INTERVENTIONS: End-tidal CO2 values were measured continuously after tracheal intubation, and were categorized as the initial value, and as minimal and maximal values during the first 20 mins. MEASUREMENTS AND MAIN RESULTS: Presenting rhythm was asystole in 22 of the first 24 patients. Return of spontaneous circulation occurred in eight patients. Initial, minimal, and maximal end-tidal CO2 values were significantly (p < .01) higher in these patients than in the patients without return of spontaneous circulation. Cutoff values providing a 100% sensitivity and the highest specificity in predicting return of spontaneous circulation were found to be 10 torr for initial and maximal end-tidal CO2 values, and 2 torr for the minimal end tidal CO2 value. The number of patients required to reject (with a risk error of <.05) the hypothesis of an actual sensitivity of < or = 90% for an observed sensitivity of 100% was found to be 95. In the second part of the study, this hypothesis was prospectively tested for initial and maximal end-tidal CO2 values in the subsequent 96 patients. Presenting cardiac rhythm was asystole in 87 patients. Return of spontaneous circulation was obtained in 30 patients. The cutoff value of 10 torr for maximal end-tidal CO2 during the first 20 mins after tracheal intubation provided an observed sensitivity of 100% in predicting return of spontaneous circulation with a specificity of 67%. This result allows rejection of the hypothesis of an actual sensitivity of < or = 90% (p = .042). By contrast, the observed sensitivity of initial end-tidal CO2 was only 87%. CONCLUSIONS: End-tidal CO2 represents a valuable tool for monitoring patients presenting with asystole during prehospital CPR. Fluctuations in end-tidal CO2 during CPR and the utility of end-tidal CO2 in detecting return of spontaneous circulation justify its continuous measurement. In addition, a high sensitivity (>90%) in predicting return of spontaneous circulation is prospectively demonstrated using the maximal end-tidal CO2 during the first 20 mins after tracheal intubation, with a cutoff value of 10 torr. Such a prognostic indicator could be used for a more rational approach to prolonged CPR. PMID- 8706456 TI - Impact of the resin blood culture medium on the treatment of critically ill patients. AB - OBJECTIVE: To assess the relevance, both clinical and bacteriologic, of the use of resin-containing blood culture media in blood cultures taken from critically ill patients receiving antibiotics. DESIGN: A prospective, open clinical trial. SETTING: The mixed medical surgical intensive care unit (ICU) of a 550-bed urban hospital. PATIENTS: All ICU patients admitted during a 3-month period (n = 49) with suspected sepsis requiring blood cultures as part of their laboratory investigations. INTERVENTIONS: The use of an aerobic resin-containing blood culture medium, in addition to the regular aerobic and anaerobic media for all blood cultures taken. MEASUREMENTS AND MAIN RESULTS: Each blood culture result was classified as to its clinical significance. Changes in patient management were recorded. Culture sets in which the resin-containing bottle provided the information central to the change in patient management were identified. Bacteriologically, the results from the resin-containing medium were compared with the results from the aerobic and anaerobic media. Of 266 blood culture sets, 103 (39%) were positive, growing 278 bacterial and fungal isolates. Clinically, the resin-containing medium alone provided relevant data leading to changes in patient management on three occasions. On two of these occasions, cultures from the regular media provided the same data within 72 hrs. Bacteriologically, 77 (29%) aerobic bottles, 55 (21%) anaerobic bottles, and 89 (33%) resin-containing bottles were positive (statistical comparison of percentages: aerobic vs. resin containing bottles, nonsignificant; aerobic vs anaerobic bottles, p < .046; anaerobic vs. resin-containing bottles, p < .0027). A similar proportion of pathogens was isolated from the resin-containing bottles only (9%) and aerobic bottles only (6%). A higher proportion of contaminants was isolated from the resin-containing bottles only than the aerobic bottles only in the various sets (17% vs. 7%, p < .046). The resin-containing bottle showed a trend toward increased detection of Staphylococcus aureus and Pseudomonas aeruginosa bacteremia. CONCLUSIONS: The resin-containing medium offers little clinical benefit to the majority of ICU patients. Bacteriologically, it seems to have a similar overall sensitivity as the regular aerobic medium (with the possible exception of a higher sensitivity for the isolation of S. aureus and P. aeruginosa), but a lower specificity. The wide-spread use of the resin-containing bottle cannot be recommended. PMID- 8706457 TI - Blood glucose and neurologic outcome with global brain ischemia. AB - OBJECTIVE: To investigate the relationship between neurologic outcome and blood glucose concentrations in survivors of cardiopulmonary arrest. DESIGN: Retrospective case series chart review. SETTING: Adult multidisciplinary intensive care unit (ICU) of a tertiary referral medical center. SUBJECTS: Consecutive patients over a 12-month period surviving cardiopulmonary resuscitation (CPR). INTERVENTIONS: Variables that were examined that could affect the relationship between the circulating glucose concentration and neurologic outcome included: location of arrest (inhospital/out-of-hospital), age, history of diabetes mellitus, duration of arrest, CPR duration, initial cardiac rhythm, and drugs administered during arrest. Cerebral recovery was evaluated by a 5-point outcome scale (Glasgow Pittsburgh Brain Stem Score) on ICU admission, and 24 and 48 hrs after ICU admission. MEASUREMENTS AND MAIN RESULTS: Observations were made on 85 patients, of whom 67% had inpatient CPR and 33% received out-of-hospital CPR. The duration of arrest of 66 (78%) patients was <5 mins. Mean CPR duration was 13.7 mins. Twenty-one percent of patients had diabetes. The mean blood glucose concentration post-CPR (n = 80) was 272 mg/dL (15.1 mmol/L). A statistically significant association was shown between high glucose concentration post-CPR and severe cerebral outcome among a small subset of patients with CPR lasting >5 min. CONCLUSIONS: The present study does not support an association between the concentration of glucose post-CPR and neurologic outcome. The previously reported casual relationship between hyperglycemia and neurologic prognosis may be an epiphenomenon of the severity of global cerebral ischemia in humans. PMID- 8706458 TI - Effect of human hemoglobin on systemic and regional hemodynamics in a porcine model of endotoxemic shock. AB - OBJECTIVE: Excessive release of nitric oxide has been implicated as being an important factor contributing to systemic arterial hypotension in septic shock. Hemoglobin is an effective nitric oxide scavenger. The purpose of this study was to test the hypothesis that treatment with cross-linked human hemoglobin can ameliorate systemic arterial hypotension and improve organ perfusion in a porcine model of normodynamic endotoxemic shock. DESIGN: Prospective, randomized, controlled trial. SETTING: Laboratory at a university medical center. SUBJECTS: Fourteen, male, random-bred swine. INTERVENTIONS: All animals were challenged with Escherichia coli lipopolysaccharide (400 microg/kg) infused from t = 0 to 90 mins. Pigs in group 1 (n = 7) were infused with cross-linked human hemoglobin (150 mg/kg) at t = 30 mins. Pigs in group 2 (n = 7) were infused at t = 30 mins with 150 mg/kg of dextran (average molecular weight 70,000 daltons) as a 5% (weight per volume) solution. MEASUREMENTS AND MAIN RESULTS: After infusion of endotoxin, mean arterial pressure decreased significantly (p < .05) but baseline cardiac index was maintained in both groups. In hemoglobin-treated pigs (group 1), mean arterial pressure was higher than in controls (group 2) from t = 60 to 120 mins (p < .05). There were no significant differences between the two groups in systemic vascular resistance index, renal blood flow, mesenteric blood flow, systemic oxygen delivery, or systemic oxygen extraction. Ileal mucosal blood flow was lower (p < .07) in group 1 than in group 2. Mean pulmonary arterial pressure increased relative to baseline in both groups, but was significantly greater in group 1 as compared with group 2. Compared with controls, infusion of hemoglobin significantly exacerbated endotoxin-induced arterial hypoxemia (p < .05). CONCLUSIONS: Treatment with hemoglobin improved mean arterial pressure in endotoxemic swine without significantly impairing blood flow to the renal or mesenteric vascular beds. Infusion of hemoglobin, however, significantly exacerbated endotoxin-induced pulmonary hypertension and arterial hypoxemia. Additional pharmacologic strategies may be necessary to ameliorate the potential adverse pulmonary effects of administering hemoglobin solutions to patients with sepsis. PMID- 8706459 TI - Decreased left ventricular contractility during porcine endotoxemia is not prevented by ibuprofen. AB - OBJECTIVE: We investigated whether ibuprofen could prevent early decrease in left ventricular contractility that occurs during porcine endotoxemia. DESIGN: Prospective, randomized, controlled animal study. SETTING: University research laboratory. SUBJECTS: Adolescent crossbred pigs (n = 28). INTERVENTIONS: Anesthetized pigs were instrumented to measure hemodynamics and left ventricular pressures (using a Millar catheter) and volumes (using a conductance catheter). Pigs were then treated in four groups, according to pretreatment using ibuprofen (15 mg/kg) or saline and subsequent treatment using endotoxin (0111:B4, 50 microg/kg) or saline. MEASUREMENTS AND MAIN RESULTS: Measurements of hemodynamics and left ventricular pressures and volumes were repeated after pretreatment with ibuprofen (or saline in controls), and at hourly intervals for 4 hrs after the start of endotoxin or control saline infusions. Left ventricular contractility was primarily assessed using the slope of the end-systolic pressure-volume relationship. Data were analyzed, using a repeated-measures analysis of variance. The slope of the end-systolic pressure-volume relationship was decreased at 4 hrs by 41 +/- 9% in the saline/endotoxin group (p < .05) and by 36 +/- 14% in the ibuprofen/endotoxin group (p < .05), so that ibuprofen pretreatment had no significant effect on the decrease in left ventricular contractility. Mean arterial pressure decreased in the saline/endotoxin group by 23 +/- 12% at 1 hr (p < .05) and by 35 +/- 12% (p < .05) at 4 hrs. Ibuprofen significantly reduced the decrease in mean arterial pressure (2 +/- 6% increased at 1 hr, and 17 +/- 12% decreased at 4 hrs, both p<.05 compared with saline/endotoxin). Cardiac output increased by 25% (p < .05) in the first hour, but then decreased to be slightly (NS) below baseline at 4 hrs in both endotoxin groups. Mean pulmonary arterial pressure was increased in the saline/endotoxin group by 154 +/- 52% (p < .05) at 30 mins and by 118 +/- 40% (p < .05) at 4 hrs. Ibuprofen prevented the very acute increase in pulmonary arterial pressure (increased by 11 +/- 33% at 30 mins, p < .05 compared with saline/endotoxin) and significantly reduced the pulmonary hypertension at 4 hrs (increased by 70 +/- 25%, p < .05 compared with both baseline and saline/endotoxin). CONCLUSIONS: We conclude that products of the cyclooxygenase pathway do not play a major role in the early decrease in left ventricular contractility after endotoxin. However, ibuprofen may have a role in reducing the other cardiovascular effects of sepsis. PMID- 8706460 TI - Granulocyte colony-stimulating factor improves survival rate and reduces concentrations of bacteria, endotoxin, tumor necrosis factor, and endothelin-1 in fulminant intra-abdominal sepsis in rats. AB - OBJECTIVE: To study the therapeutic effect of granulocyte colony-stimulating factor (G-CSF) on the mortality rate and host defense pattern in fulminant intra abdominal sepsis. DESIGN: Prospective, randomized, controlled trial. SETTING: Research laboratory in a university hospital. SUBJECTS: Adult male Wistar rats. INTERVENTIONS: Fulminant polymicrobial intra-abdominal sepsis was induced by a 4 mm cecal perforation. Survival experiments were performed with two different doses of G-CSF (20 and 100 microg/kg/24 hrs), and therapy was started 7 days or 1 day before, or 4 hrs after sepsis induction (n = 24). To examine alterations in host response pattern, G-CSF (20 microg/kg/24 hrs) was given at sepsis induction, and rats were killed 4, 8, 12 and 24 hrs later (n = 8-16 per time period). Histologic examination of lung, liver, spleen, and kidney was performed, and blood concentrations of bacteria, endotoxin, tumor necrosis factor (TNF), endothelin-1, packed cell volume, and lactate were determined. MEASUREMENTS AND MAIN RESULTS: G-CSF (20 microg/kg/24 hrs), given 4 hrs after sepsis induction, reduced the mortality rate from 96% to 42%. Increasing the dose (100 micrograms/kg/24 hrs), or giving G-CSF as prophylaxis (starting 7 days or 1 day before sepsis), gave no further protection. G-CSF attenuated the sepsis-induced enhancement of circulating bacteria, endotoxin, TNF, and endothelin-1, resulting in improved fluid balance and reduced lactate concentration. No histopathologic alterations were observed after G-CSF treatment. CONCLUSIONS: G-CSF improves host defense and survival rate in experimentally induced fulminant intra-abdominal sepsis. Clearance of bacteria and endotoxin is improved, concentrations of TNF and endothelin-1 are suppressed, and microvascular flow is improved. G-CSF does not induce neutrophil-mediated tissue damage. PMID- 8706461 TI - Correction of blood pH attenuates changes in hemodynamics and organ blood flow during permissive hypercapnia. AB - OBJECTIVES: To determine whether changes in cardiac output, regional blood flow, and intracranial pressure during permissive hypercapnia are blood pH-dependent and can be attenuated by correction of intravascular acidemia. DESIGN: Prospective, controlled study. SETTING: Research laboratory. SUBJECTS: Female Marino ewes. INTERVENTIONS: Animals were instrumented with a pulmonary artery catheter, femoral arterial and venous catheters, a catheter in the third cerebral ventricle, and ultrasonic flow probes on the left carotid, superior mesenteric, and left renal arteries 1 wk before experimentation. At initiation of the protocol, ewes underwent endotracheal intubation and mechanical ventilation under general anesthesia. Minute ventilation was reduced to induce hypercapnia with a target PaCO2 of 80 torr (10.7 kPa). In the pH-uncorrected group (n = 6), arterial blood pH was allowed to decreased without treatment. In the pH-corrected group (n = 5), 14.4 mEq/kg of sodium bicarbonate was given intravenously as a bolus to correct arterial blood pH toward a target arterial pH of 7.40 (dose calculated by the Henderson-Hasselbalch equation). MEASUREMENTS AND MAIN RESULTS: Arterial blood pH, PCO2, cardiac output, intracranial pressure, and carotid, superior mesenteric, and renal artery blood flow rates were measured at normocapnic baseline and at every hour during hypercapnia for 6 hrs. In the pH-uncorrected group, arterial blood pH decreased from 7.41 +/- 0.03 at normocapnia to 7.14 +/- 0.01 (p < .01 vs. normocapnia) as blood PCO2 increased to 81.2 +/- 1.8 torr (10.8 +/- 0.2 kPa). In the pH-corrected group, arterial blood pH was 7.42 +/- 0.02 at normocapnia and was maintained at 7.37 +/- 0.01 while PaCO2 was increased to 80.3 +/- 0.9 torr (10.7 +/- 0.1 kPa). Significant increases in cardiac output occurred with the initiation of hypercapnia for both groups (pH-uncorrected group: 4.3 +/- 0.6 L/min at normocapnia vs. 6.8 +/- 1.0 L/min at 1 hr [p < .05]; pH-corrected group: 4.1 +/- 0.4 at normocapnia vs. 5.7 +/- 0.4 L/min at 1 hr [p < .05]). However, this increase was sustained only in the uncorrected group. Changes in carotid and mesenteric artery blood flow rates, as a percent of baseline values, showed sustained significant increases in the pH-uncorrected groups (p < .05) and only transient (carotid at 1 hr) or no (superior mesenteric) significant change in the pH-corrected groups. Conversely, significant increases in renal artery blood flow were seen only in the pH-uncorrected group during the last 2 hrs of the experiment (p < .05). Organ blood flow, as a percent of cardiac output, did not change significantly in either group. Intracranial pressure increased significantly in the pH-uncorrected group (9.0 +/- 1.5 mm Hg at normocapnia vs. 26.8 +/- 5.1 at 1 hr, p < .05), and remained increased, while showing no significant change in the pH-corrected group (8.5 +/- 1.6 mm Hg at normocapnia to 7.7 +/- 4.2 at 1 hr). CONCLUSIONS: Acute hypercapnia, induced within 1 hr, is associated with significant increases in cardiac output, organ blood flow, and intracranial pressure. These changes can be significantly attenuated by correction of blood pH with the administration of sodium bicarbonate, without adverse effects on hemodynamics. PMID- 8706463 TI - Nitric oxide inhalation decreases pulmonary platelet and neutrophil sequestration during extracorporeal circulation in the pig. AB - OBJECTIVE: The inhibiting effect of nitric oxide on the aggregation and adhesion of neutrophils and platelets has been well documented in vitro. In vivo evidence, however, is more scant. In this study, we studied the effects of inhaled nitric oxide on pulmonary cellular sequestration in our sham hemodialysis model. Accumulation of neutrophils and platelets in the lungs has been shown to be an early event in this model. DESIGN: Prospective, randomized, controlled study. SETTING: Animal laboratory at a university medical center. SUBJECTS: Twenty-six anesthetized, mechanically ventilated pigs. INTERVENTIONS: 111Indium-oxine was used to selectively label neutrophils or platelets and the activity over the lungs was followed dynamically with a gamma camera. Sham hemodialysis, using a cuprophan hollow-fiber dialyzer, was instituted via catheters in the femoral vessels. The animals were divided into two main groups: a) the nitric oxide recipient group (n = 12, with platelets labeled in seven animals and neutrophils labeled in five animals); and b) the control group (n = 14, with platelets labeled in seven animals and neutrophils labeled in seven animals). The animals in the former group were given 50 parts per million of nitric oxide in the inspiratory gas from the beginning of dialysis and for 30 mins onward. MEASUREMENTS AND MAIN RESULTS: Inhalation of nitric oxide attenuated the increase in activity over the lungs in both the neutrophil and platelet groups during sham hemodialysis. In addition, an inhibiting effect on the increase in pulmonary pressure was noted. CONCLUSION: Apart from the effects of nitric oxide on central hemodynamics in this model, the scintigraphic findings indicate an in vivo effect of nitric oxide on the accumulation of platelets and neutrophils in the lungs, probably due to inhibition of the adhesion and/or aggregation of these cells. PMID- 8706462 TI - Nitric oxide synthase inhibition versus norepinephrine for the treatment of hyperdynamic sepsis in sheep. AB - OBJECTIVES: To investigate the effects of Nomega-mono-methyl-L-arginine (L-NMMA), an inhibitor of nitric oxide synthesis, on hemodynamics, oxygen transport, and regional blood flow in an ovine model of hyperdynamic sepsis and to compare these effects with the responses to norepinephrine. DESIGN: Prospective, nonrandomized, controlled experimental study with repeated measures. SETTING: Investigational intensive care unit at a university medical center. SUBJECTS: Twenty-five female, healthy, adult sheep of the Merino breed, divided into three groups: nine control sheep; eight sheep treated with L-NMMA; and eight sheep treated with norepinephrine. INTERVENTIONS: All sheep were chronically instrumented. After a 5 day recovery period, a continuous infusion of live Pseudomonas aeruginosa (2.5 x 10(6) colony-forming units/min) was started and maintained for the remainder of the experiment. After 24 hrs of sepsis, eight sheep received L-NMMA (7 mg/kg/hr), eight sheep received norepinephrine, and nine sheep received the vehicle alone (0.9% saline). The norepinephrine dosage was continuously and individually adjusted to achieve the same increase in blood pressure as was observed in a matched sheep of the L-NMMA group. MEASUREMENTS AND MAIN RESULTS: After 24 hrs of sepsis, all sheep developed a hyperdynamic circulatory state with increased cardiac indices and reduced arterial pressures, and systemic vascular resistances. L-NMMA reversed the hyperdynamic circulation, causing an increase in arterial pressure by peripheral vasoconstriction. Norepinephrine led to an increase in blood pressure by augmenting cardiac indices, leaving the systemic vascular resistance unaffected. The norepinephrine dose needed to keep the blood pressure high had to be continuously increased, reflecting the reduced vascular responsiveness to catecholamines during sepsis. Renal blood flow remained unaffected by all treatment forms. Norepinephrine and L-NMMA led to a dramatic increase in urine production. Blocking the nitric oxide synthase with L-NMMA did not interfere with the host's pulmonary ability to clear bacteria, nor did treatment with norepinephrine. CONCLUSIONS: Blocking nitric oxide synthase had a marked vasoconstrictive effect. Both norepinephrine and L-NMMA increased arterial pressure without reducing renal blood flow, leading to an improved renal function. PMID- 8706464 TI - Effect of sepsis and 3,5,3'-triiodothyronine replacement on myocardial integrity during oxidant challenge. AB - OBJECTIVE: To determine whether sepsis, with or without thyroid hormonal augmentation, induces myocardial tolerance to an oxidant challenge. DESIGN: A prospective, randomized, controlled animal trial. SETTING: University research laboratory. SUBJECTS: Twenty male Sprague-Dawley rats. INTERVENTIONS: After anesthesia, animals underwent cecal ligation and puncture, with or without 3,5,3' triiodothyronine replacement (3 ng/hr), or sham surgery. Twenty-four hours later, the heart was rapidly excised for retrograde Langendorff perfusion. Oxyradical challenge consisted of the addition of 200 microM of hydrogen peroxide to the perfusate for 60 mins. MEASUREMENTS AND MAIN RESULTS: Myocardial contractility and relaxation were continuously recorded. Perfusate glutathione and lactate dehydrogenase concentrations were determined enzymatically at 30-min intervals for 90 mins. Oxyradical perfusion alone significantly increased glutathione efflux and decreased myocardial contractility when compared with control animals. Prior cecal ligation and puncture decreased oxidant-mediated glutathione efflux and maintained myocardial contractility. 3,5,3'-triiodothyronine supplementation appeared to increase late cardiac contractility and cellular integrity during oxidant challenge. However, this increase was not statistically significant. CONCLUSIONS: Antecedent septic challenge appears to induce tolerance to further myocardial oxyradical exposure and improves myocardial functional and biochemical integrity. Thyroid hormonal supplementation may provide a modest additional benefit in septic animals. PMID- 8706465 TI - Evaluation of the effects of autotransfusion of unprocessed blood on hemodynamics and oxygen transport in anesthetized pigs. AB - OBJECTIVE: To examine the hemodynamic effects and the oxygen transport pattern of autotransfusion of unprocessed blood on hemodynamics and oxygen transportation. DESIGN: Prospective, observational study. SETTING: Research laboratory of a university medical center. SUBJECTS: Six healthy, domestic pigs (20 - 33 kg). INTERVENTIONS: A left thoracotomy was performed and a 5-mm incision was created in the descending aorta, resulting in a controlled hemorrhage of 30 mL/kg (approximately 40% of blood volume) into the thoracic cavity over a 45-min time period. During that period, mean arterial pressure (MAP) was maintained slightly > 50 mm Hg, using intravenous lactated Ringers' solution. The blood sample was collected from the open thorax with compresses soaked in citric acid solution and then extracted by manual squeezing, filtered through several layers of gauzes, and stored in glass bottles. Repeat measurements were performed after hemorrhage, after retransfusion of the harvested blood, and thereafter every 15 mins up to 60 mins. The animals were supported for 2 more hrs and were observed for the following 48 hrs. MEASUREMENTS AND MAIN RESULTS: All animals survived and were in good condition 48 hrs after the experimental hemorrhage. The circulatory and oxygen transport response at the end of hemorrhage and concomitant maintenance of blood pressure at > 50 mm Hg resulted in: significant reductions of cardiac index, MAP, and oxygen transport (DO2) (46%, 50%, and 64% reductions, respectively, p < .01, in an increase of heart rate (HR) (+21%, not significant), pulmonary vascular resistance index (+112%, p < .05), and oxygen extraction (+105%, p < .01), as well as in a nonsignificant decrease of systemic vascular resistance index (-8%). After autotransfusion, the basic hemodynamic variables, MAP and HR were corrected, remaining near baseline (not significant) afterward. Cardiac index and DO2 increased after autotransfusion, but remained below the baseline until the end of the study protocol (p < .05). A significant increase was noticed for pulmonary arterial pressure and pulmonary vascular resistance index immediately after autotransfusion (p < .01). These values were corrected in part after 15 to 30 mins, but remained higher throughout the observaton period compared with baseline (29.5% and 89.8%, respectively, p < .05). The recently introduced relationship between cardiac index and oxygen extraction has been proposed to avoid problems of mathematical coupling between oxygen consumption and DO2 measurements. This relationship followed a similar course in all experiments throughout each phase. A shift downward and to the right represented the endpoint of the hemorrhagic phase. After autotransfusion, a shift toward baseline was noticed. Prothrombin time and partial thromboplastin time remained unchanged after autotransfusion. Free hemoglobin concentrations increased immediately after autotransfusion (+33%, p < .05), but returned to baseline values 48 hrs later. Histologic examination showed no changes in the examined organs. CONCLUSIONS: Reinfusion of large amounts of unprocessed blood (up to 40% of blood volume), collected with compresses from a noncontaminated surgical field is a cheap method, which may be of potential benefit in trauma patients, when more sophisticated autotransfusion devices are lacking. In the present study, this method resulted in transient but significant hemodynamic changes in the pulmonary circulation. Impairment of oxygen transport was noticed after the end of hemorrhage, but this impairment cannot be attributed to the autotransfusion technique alone, but also to factors such as hemorrhagic shock, surgical trauma, etc. PMID- 8706466 TI - Inspired oxygen concentration alters the phospholipids and protein content in the bronchoalveolar lavage-accessible space. AB - OBJECTIVE: To examine the effect of FIO2 on the contents of total protein, total phospholipids, phosphatidylcholine, and phosphatidylglycerol in the bronchoalveolar lavage-accessible space in male and female rats in vivo. DESIGN: Prospective, controlled trial. SETTING: Research laboratory. SUBJECTS: Adult male and female Sprague-Dawley rats. INTERVENTIONS: After animals were anesthetized with an intraperitoneal injection of pentobarbitol (50 mg/kg), a 24-gauge catheter was placed in the femoral artery. Determinations of arterial pH and PaO2 and PaCO2 were performed before tracheostomy, and all animals were then ventilated for 3 mins with an FIO2 of 0.21, followed by a reduction bronchoalveolar lavage. The animals were randomly divided equally by gender and given either an FIO2 of 0.21, 0.50, or 1.00. All subjects were ventilated in the same manner. Sampling bronchoalveolar lavage was performed 80 and 160 mins after institution of the variable FIO2. Bronchoalveolar lavage samples were analyzed for protein and phospholipid content. Arterial blood was obtained for determination of arterial pH and the PaO2 and PaCO2 immediately and 40 mins after each sampling bronchoalveolar lavage. MEASUREMENTS AND MAIN RESULTS: At the times of bronchoalveolar sampling lavage, the PaCO2 increased and the PaO2 decreased, as did the pH. In the 40-min samples obtained between sampling lavages, the arterial pH and PaCO2 and PaO2 returned to pretracheostomy values (animals ventilated with an FIO2 of 0.21) and/or higher pO2 values (animals ventilated with an FIO2 of 0.5 or 1.0). No differences were found between genders in amounts of total protein and phospholipid content in reduction and zero time bronchoalveolar lavage fluid. Males and females ventilated with an FIO2 of 0.21 differed in the amounts of total protein, total phospholipids, phosphatidylcholine, and phosphatidylglycerol found in sampling bronchoalveolar lavage at 80 and 160 mins. Amounts of total protein and total phospholipids also demonstrated gender differences with the administration of an FIO2 of 1.0, but no differences with the administration of and FIO2 of 0.5. In terms of the phospholipids, males had greater amounts in the sampling bronchoalveolar lavage at 80 mins, and females at 160 mins. Administration of an FIO2 of 0.5 or 1.0 resulted in decreased amounts of total phospholipids in both males and females when compared with and FIO2 of 0.21. In males, administration of both FIO2 of 0.5 and 1.0 resulted in decreased amounts of phosphtidylcholine found in the bronchoalveolar lavage-accessible space; in females, amounts of phosphatidylcholine were only decreased when and FIO2 of 1.0 was administered. In males, administration of and FIO2 of 1.0 also resulted in decreased amounts of phosphatidylglycerol. The decreased amount of phosphatidylglycerol occurred in females given an FIO2 of 0.5. Amounts of total protein in males and females given an FIO2 of 0.5 and in females given an FIO2 of 1.0 were found to be increased. CONCLUSIONS: Our findings support the hypothesis that hyperoxia alters surfactant composition. Further investigation is warranted to determine the mechanisms affecting secretion of phosphatidylcholine and phosphatidylglycerol into the bronchoalveolar space and to explore the gender difference in secretion. PMID- 8706467 TI - Albuterol delivery by metered-dose inhaler in a mechanically ventilated pediatric lung model. AB - OBJECTIVES: To assess albuterol delivery by metered-dose inhaler in a mechanically ventilated pediatric lung model and to determine the influence of the following variables on albuterol delivery: endotracheal tube diameter; type of spacer; humidification; and pulmonary mechanics. DESIGN: Prospective, in vitro, laboratory study. SETTING: Research laboratory. INTERVENTIONS: A model, consisting of a volume-cycled ventilator, pediatric breathing circuit, 4.0- or 6.0-mm endotracheal tube, and lung simulator, was assembled. Ventilator settings were: tidal volume 250 mL; FIO2 0.5; inspiration/expiration ratio 1:3; respiratory rate 25 breaths/min; positive end-expiratory pressure 3 cm H2O; temperature 35 degrees C; and a decelerating flow pattern, using dry and humidified air. Lung simulator compliance and resistance values were consistent with those values reported for healthy childeren (20 mL/cm H2O and 40 cm H20/L/sec) and children with pulmonary disease (10 mL/cm H2O and 60 cm H2O/L/sec). Pulmonary mechanics were verified with a pulmonary function diagnostic system. Ten metered-dose inhaler canisters were used to administer 2000 micrograms of albuterol, using either a collapsible or a rigid spacer. A circuit filter placed immediately proximal to the test lung collected drug exiting the endotracheal tube. The filter was rinsed with water and albuterol concentrations were determined by high-performance liquid chromatography. Each variable was tested in triplicate. MEASUREMENTS AND MAIN RESULTS: Albuterol delivery was significantly (p < or = .05) greater for the 6.0-mm endotracheal tube, rigid spacer, dry air, and pulmonary disease mechanics by multifactor analysis of variance. Drug delivery in humidified air with pulmonary disease mechanics using the rigid chamber was 2.5 =/- 0.27% and 6.3 =/- 0.99% for the 4.0 and 6.0-mm endotracheal tubes, respectively. CONCLUSIONS: These in vitro results suggest that pulmonary disease mechanics and a 6.0-mm endotracheal tube improve albuterol delivery. Future clinical investigations in intubated pediatric patients with pulmonary disease are needed to address the clinical significance of these results. PMID- 8706468 TI - Risk factors for nosocomial infection in critically ill children: a prospective cohort study. AB - OBJECTIVE: To identify factors in pediatric intensive care unit (ICU) patients that are associated with an increased risk of nosocomial infections. DESIGN: A prospective, 1-yr cohort study. SETTING: A 16-bed pediatric ICU in a multidisciplinary, regional referral center. SUBJECTS: All patients admitted to the pediatric ICU. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: The primary outcome variable was the development of nosocomial infection. Out of 945 consecutive admissions, 75 patients developed 96 nosocomial infections. The most frequent infection sites were the lower respiratory tract (35%), the bloodstream (21%), and the urinary tract (21%). The most common organisms isolated were Gram negative bacteria (53%, Gram-positive bacteria (27%), and fungi (9%). Variables significantly associated with the development of nosocomial infections included age, weight, Pediatric Risk of Mortality (PRISM) score, device utilization ratio, antimicrobial therapy, histamine-2 (H2) receptor blocker use, immune status, parenteral nutrition, and length of stay. When combined in a multivariate logistic regression model, the significant variables were operative status, PRISM score, device utilization ratio, antimicrobial therapy, parenteral nutrition, and length of stay before the onset of infection. The area under the receiver operating characteristic curve was 0.868. At a probability of 0.15, the sensitivity was 66.67%, and the specificity was 87.82%. CONCLUSIONS: Patients at risk for developing nosocomial infection can be identified using a multivariate logistic regression model with a high degree of sensitivity and specificity. These data indicate that institutional nosocomial rates need to be adjusted for risk factors. This model could help target patients at high risk for developing nosocomial infections for preventive strategies. PMID- 8706469 TI - Accuracy and reliability of disposable pressure transducers coupled with modern pressure monitors. AB - OBJECTIVE: To determine the bedside accuracy of direct patient pressure monitoring when used with new and clinically used disposable blood pressure (BP) transducers. DESIGN: Prospective study. SETTING: Laboratory bench and critical care units in an adult and children's hospital. SUBJECTS: Seventy-five bedside patient monitors (25 Marquette Electronics, 25 Spacelab Medical, and 25 Hewlett Packard), and 100 disposable transducers (50 from Utah Medical Products and 50 from Abbott Critical Care Systems [25 new, 25 clinically used of each manufacturer]) were tested. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: A +/- 2% accuracy requirement for bedside monitors and the +/- 3% American National Standards Institute accuracy standard for disposable BP transducers were used. To test the accuracy of the bedside monitors, a certified transducer simulator was used to apply 100 mm Hg to each bedside monitor. To test the accuracy of the disposable BP transducers, a very accurate (+/- 0.05%) pneumatic dead weight tester was used to apply pressures to the transducer. A digital power supply and a 6 1/2 digit voltmeter were used. The average output of the bedside monitors when 100 mm Hg was applied was 99.90 +/- 0.83 mm Hg, with the worst cases being 98 and 103 mm Hg. For all 100 disposable pressure transducers, the average output was 100.03 +/- 0.55 mm Hg, with the worst cases being 98.53 and 101.36 when 100 mm Hg was applied. There was no important difference in the accuracy of the transducers obtained from the two vendors nor whether the transducers had been used clinically. CONCLUSIONS: All disposable BP transducers tested were much more accurate than the American National Standards Institute standard for accuracy. Even the worst case transducers were twice as accurate as required by the American National Standards Institute standard. Only one bedside monitor was outside the +/- 2% accuracy range (103 mm Hg). Based on these findings, this author recommends that fixed calibration disposable transducers and fixed calibration bedside pressure monitoring systems be used. The clinical risks of air embolism and infection from the calibrating mercury manometer and the complexity of the calibration task are the overriding factors for making these recommendations. PMID- 8706470 TI - Can healthcare providers obtain judicial intervention against surrogates who demand "medically inappropriate" life support for incompetent patients? AB - OBJECTIVE: This article analyzes, from a legal perspective, a recent phenomenon involving a clash between the values of attending medical personnel and the instructions of surrogate decision-makers acting on behalf of incompetent patients. Some hospitals have gone to court to challenge decisions by surrogates to continue life support for permanently unconscious or other gravely debilitated patients. Their claim has been that continuation of life support would be medically inappropriate and that the surrogates' decisions ought to be overridden. These petitions have thus far been rejected. The objective here is to explain those decisions and to predict the outcome of future, similar litigation. DATA SOURCES: The primary data are the judicial decisions and legislation accumulated since the Quinlan case in 1976, regarding the medical handling of dying medical patients. CONCLUSIONS: Judicial rejection of healthcare providers' claims in the decided cases is explainable under traditional guardianship principles. The explanation lies in surrogates' authority to make decisions in the best interests of incompetent patients, and in judicial reluctance to brand life preservation of nonsuffering patients as abusive or contrary to patient interests. At the same time, the author anticipates a change in judicial posture, as courts acknowledge the widespread antipathy of people toward being indefinitely preserved in a noncognitive status. Because the judicial approach to the handling of dying persons often seeks to replicate what the patient would have wanted, there is room to consider consensus preferences where the particular patients has never indicated any deviation from those preferences. Courts will eventually override surrogate decisions that do not conform to widely shared preferences for avoiding the indignity of permanent unconsciousness or other gravely debilitated states. PMID- 8706471 TI - The American Board of Internal Medicine Recertification Program - nearing liftoff. PMID- 8706472 TI - Critical care in 1996: Doing too much? Doing too little? Keeping the patient in focus during a time of smoke and fire. The presidential address from the 25th Educational and Scientific Symposium of the Society of Critical Care Medicine. PMID- 8706473 TI - Potentially lethal misplacement of femoral central venous catheters. PMID- 8706474 TI - Head injury management in the United States. PMID- 8706475 TI - Age as a component of severity of illness scoring. PMID- 8706476 TI - Steroid replacement in sepsis: an unexplored side of a multifaceted drug class. PMID- 8706477 TI - Ventilator-associated pneumonia. PMID- 8706478 TI - Surfactant replacement in pediatric respiratory failure: promising therapy for the future? PMID- 8706479 TI - The expanding spectrum of critical illness polyneuropathy. PMID- 8706480 TI - Results of the Multidisciplinary Critical Care Knowledge Assessment Program, 1996. PMID- 8706481 TI - Nuclear factor-kappa B is activated in alveolar macrophages from patients with acute respiratory distress syndrome. AB - OBJECTIVE: The expression of proinflammatory cytokines is rapidly increased in experimental models of the acute respiratory distress syndrome (ARDS), in patients at risk for ARDS, and in patients with established ARDS. Because multiple cytokines are present in bronchoalveolar lavage fluid, a common, proximal activation mechanism may operate in these settings. The proinflammatory cytokines whose expression is increased in the lungs of patients with ARDS have binding sequences in their enhancer/promoter regions for transcriptional regulatory proteins, such as nuclear factor-kappa B (NF-kappa B), nuclear factor IL6 (NF-IL6), cyclic adenosine monophosphate responsive element binding protein, serum protein-1, and activating protein-1. To test the hypothesis that activation of one or more of these nuclear transcriptional regulatory factors might provide a common mechanism for the simultaneous expression of multiple cytokine genes in the setting of ARDS, we measured activation of these factors in alveolar macrophages from patients with ARDS and from controls. DESIGN: Prospective, clinical study. SETTING: Medical and surgical intensive care units at a university hospital and a county hospital. PATIENTS: Twelve patients, six with established ARDS and six control patients without lung injury. INTERVENTIONS: Patients with ARDS and controls underwent fiberoptic bronchoscopy and bronchoalveolar lavage. Alveolar macrophages were isolated from lavage fluid and the nuclear proteins were extracted. Activation of transcriptional factors NF kappa B, NF-IL6, cyclic adenosine monophosphate responsive element binding protein, activating protein-1, and serum protein-1 was determined using an electrophoretic mobility shift assay, followed by densitometry of the autoradiographed gels. MEASUREMENTS AND MAIN RESULTS: There were no significant differences in gender, age, tobacco smoking, Acute Physiology and Chronic Health Evaluation II score, quantity of lavage fluid, or number of alveolar macrophages in lavage specimens in the patient groups. Acute Lung Injury score and the Pao2/Fio2 ratio differed significantly between controls and ARDS patients: 0.46 +/- 0.17 vs. 2.74 +/- 0.14 (p < .0001) and 310 +/- 45 torr (41.3 +/- 6.0 kPa) vs. 150 +/- 11 torr (21.3 +/- 1.5 kPa) (p < .006), respectively. The mean Fio2 of the control patients was not significantly different from the mean Fio2 of ARDS patients: 0.47 +/- 0.11 vs. 0.55 +/- 0.6 (p = .53). Patients with ARDS had significantly (p < .02) increased activation of NF-kappa B in alveolar macrophages compared with patients without the syndrome. There was no evidence of increased activation of the transcriptional factors activating protein-1, serum protein-1, NF-IL6, or cyclic adenosine monophosphate responsive element binding protein in alveolar macrophages from ARDS vs. control patients. CONCLUSIONS: These experiments demonstrated increased in vivo activation of the nuclear transcriptional regulatory factor NF-kappa B (but not NF-IL6, cyclic adenosine monophosphate responsive element binding protein, activating protein-1, or serum protein-1) in alveolar macrophages from patients with ARDS. Because binding sequences for NF-kappa B are present in the enhancer/promoter sequences of multiple proinflammatory cytokines, activation of NF-kappa B may contribute to the increased expression of multiple cytokines in the lung in the setting of established ARDS. PMID- 8706482 TI - Perioperative endotoxemia and bacterial translocation during major abdominal surgery: evidence for the protective effect of endogenous prostacyclin? AB - OBJECTIVE: To investigate the potential role of endogenous prostacyclin (PGI2) released after mesenteric traction during major abdominal surgery on perioperative endotoxemia and bacterial translocation. DESIGN: Prospective, randomized, double-blind clinical study. SETTING: Operating room and surgical intensive care unit in a university hospital. PATIENTS: Fifty consecutive patients scheduled for major abdominal surgery (pancreas resection, abdominal aortic surgery). INTERVENTIONS: Fifteen minutes before skin incision, either 400 mg of ibuprofen or a placebo equivalent were administered intravenously. Immediately after peritoneal incision, eventration and action of the small bowel was intentionally performed in a uniform fashion. MEASUREMENTS AND MAIN RESULTS: Baseline values were obtained before induction of anesthesia. Additional measurements, along with assessments of hemodynamics and gas exchange, were performed before incision of the peritoneum and at 5, 30, and 45 mins and 3, 6, and 24 hrs after mesenteric traction. Arterial plasma concentrations of 6-keto prostaglandin F1 alpha and thromboxane B2 (stable metabolites of PGI2 and thromboxane A2) were determined by radioimmunoassay. Endotoxin was measured by limulus amebocyte lysate test. Mesenteric lymph nodes were sampled in 31 patients (ibuprofen n = 14, placebo n = 17) and sent for culture under sterile conditions. Transient hypotension and a marked increase of plasma 6-keto-prostaglandin F1 alpha concentrations occurred up to 6 hrs after mesenteric traction in untreated patients with median peak concentrations (2243 vs. 72 ng/L [p < .0001, placebo vs. ibuprofen], observed 5 mins after mesenteric traction). Endotoxemia occurred in both study groups. However, after mesenteric traction, plasma endotoxin concentrations were significantly higher in the ibuprofen group. Median peak concentrations (0.12 vs. 0.27 EU/mL [p < .001, placebo vs. ibuprofen]) were observed 3 hrs after mesenteric traction. Gram-negative bacteria in mesenteric lymph nodes were detected exclusively in the ibuprofen group (n = 5, p < .01). CONCLUSIONS: In ibuprofen-pretreated patients, significantly higher endotoxin concentrations as well as bacterial translocation to mesenteric lymph nodes occurred, despite the absence of a transient decrease in mean arterial pressure that had been associated with PGI2 release. Therefore, we hypothesized that during major abdominal surgery, endogenous PGI2 released in response to mesenteric traction may play a crucial role in maintaining splanchnic microcirculation and thus preserving gut mucosal barrier function. PMID- 8706483 TI - N-acetyl-L-cysteine depresses cardiac performance in patients with septic shock. AB - OBJECTIVE: To investigate the effects of adjunctive therapy with parenteral N acetyl-L-cysteine in patients with newly diagnosed septic shock. DESIGN: Prospective, randomized, double-blind, placebo-controlled study. SETTING: Multidisciplinary intensive care unit at a university teaching hospital. PATIENTS: Twenty patients (N-acetyl-L-cysteine group [n = 10], placebo group [n = 10]), 15 male and five female, of mean age 64 +/- 15 (SD) yrs and Acute Physiology and Chronic health Evaluation (APACHE) II score 33 +/- 6, with septic shock within 24 hrs of diagnosis. INTERVENTIONS: After a 2-hr stabilization period (time-zero minus 2 hrs to time-zero), patients received either N-acetyl-L cysteine in 5% dextrose (150 mg/kg in 100 mL over 15 mins, followed by 50 mg/kg in 250 mL over 4 hrs, and then 100 mg/kg/24 hrs in 500 mL for 44 hrs; N-acetyl-L cysteine group) or the equivalent volume of 5% dextrose (placebo group). MEASUREMENTS AND MAIN RESULTS: Hemodynamic and oxygen transport indices were measured at time-zero minus 2 hrs and time-zero, and at multiple time points thereafter until completion of the trial infusion (time-zero plus 48 hrs). A daily Organ Failure Score was recorded for 14 days. Treatment group demographics and hemodynamic variables did not differ significantly between the two groups at time-zero. Mean (SD), pooled mean arterial pressure (MAP), and cardiac index were 75 +/- 15 mm Hg and 3.9 +/- 1.2 L/min/m2, respectively. Over the next 48 hrs, in the N-acetyl-L-cysteine group, there was a progressive decrease, relative to both time-zero and the placebo group, in MAP, cardiac index, and left ventricular stroke work index (p < .01, repeated-measures analysis of variance). Percentage reductions in these values relative to the placebo group at 48 hrs were 23%, 18%, and 43%, respectively Oxygen transport indices, arterial blood gas analyses, Pao2/Fio2 ratio, and shunt did not differ over time between the groups. There was no difference in either daily Organ Failure Score over time (p > .01, repeated measures analysis of variance) or hospital mortality rate (90% N-acetyl-L cysteine group, 50% placebo group) (p > .1, logistic regression) between the two groups. CONCLUSION: Adjunctive therapy with N-acetyl-L-cysteine in newly diagnosed septic shock was associated with a depression in cardiovascular performance, as indicated by progressive reductions in cardiac index, left ventricular stroke work index, and MAP. PMID- 8706484 TI - Administration of amphotericin B in lipid emulsion decreases nephrotoxicity: results of a prospective, randomized, controlled study in critically ill patients. AB - OBJECTIVES: To evaluate the differences in efficacy and in clinical and biochemical tolerance to amphotericin B administered in a lipid emulsion compared with amphotericin B administered in 5% dextrose in water in the treatment of Candida albicans infection in intensive care unit (ICU) patients. DESIGN: Prospective, controlled, randomized study, conducted during a 2.5-yr period, comparing the two treatment protocols. SETTING: General ICU of a university affiliated municipal hospital. PATIENTS: Sixty consecutive critically ill patients with confirmed or suspected Candida infection. INTERVENTIONS: Patients received amphotericin B (1 mg/kg/24 hrs), administered randomly in 5% dextrose in water (group A), or in lipid emulsion (20% intralipid) (group B). MEASUREMENTS AND MAIN RESULTS: Clinical tolerance (fever, chills, hemodynamics), hepatorenal tolerance, and biological tolerance (serum electrolytes and coagulation profile) were evaluated. Patients receiving amphotericin B in lipid emulsion experienced a lower frequency rate of drug-associated fever (61.4% vs. 5.8%, p < .003) rigors (54% vs. 8.5%, p < .004), hypotension (17% vs. 0%), and nephrotoxicity (increase of serum creatinine concentration 66.7% vs. 20%, p < .0002). Significant (264,500 +/- 71,460 to 163,570 +/- 34,450 mm3, p < .01) thrombocytopenia, not associated with active bleeding, occurred in patients receiving amphotericin B lipid in emulsion but not in patients receiving the drug in dextrose. CONCLUSIONS: Treatment with amphotericin B in a lipid emulsion when given to critically ill patients with Candida sepsis seems to be safer and as effective as the conventional mode of administration. PMID- 8706485 TI - Calf's lung surfactant extract in acute hypoxemic respiratory failure in children. AB - OBJECTIVE: Open-label trial of the safety and short-term efficacy of calf's lung surfactant in pediatric respiratory failure. DESIGN: Multi-institutional, uncontrolled, observational trial. SETTING: Six pediatric intensive care units of tertiary medical centers. PATIENTS: Twenty-nine children with acute hypoxemic respiratory failure, characterized by diffuse, bilateral, pulmonary infiltrates, need for ventilator support, and an oxygenation index of > or = 7. INTERVENTIONS: Up to four doses of intratracheal surfactant (80 mL/m2). MEASUREMENTS AND MAIN RESULTS: Ventilator parameters, arterial blood gases, and derived oxygenation and ventilation indices were recorded before, and at intervals after, surfactant administration. Complications and outcome measures were also noted. There was immediate improvement in oxygenation and moderation of ventilator support associated with surfactant administration in 24 of 29 patients. A modest but statistically insignificant effect was seen with subsequent doses. The only complications occurred in three patients who developed airleaks, two of which were coincident with surfactant administration. The overall mortality rate was 14%, which compares favorably with other published series. CONCLUSIONS: Administration of calf's lung surfactant appears to be safe and is associated with rapid improvement in oxygenation and moderation of ventilator support in children with acute hypoxemic respiratory failure. These results set the stage for a randomized, controlled study. PMID- 8706486 TI - Polysomnography after adenotonsillectomy in mild pediatric obstructive sleep apnea. AB - OBJECTIVES: a) To determine the need for intensive monitoring on the first operative night of surgery in children undergoing adenotonsillectomy for mild obstructive sleep apnea; b) to examine the effect of narcotics on postoperative obstructive sleep apnea. DESIGN: Randomized, prospective study. SETTING: University hospital. PATIENTS: Children, ranging in age between 1 and 18 yrs, presented to the Pediatric Otolaryngology Clinic for adenotonsillectomy for mild obstructive sleep apnea defined as from one to 15 obstructive apnea events per hour on preoperative polysomnogram. INTERVENTIONS: Patients were assigned to receive either a narcotic- or a halothane-based anesthetic for adenotonsillectomy. A postoperative polysomnogram was performed in the pediatric intensive care unit on the first operative night. MEASUREMENTS AND MAIN RESULTS: Eighteen patients were recruited, 15 of whom met inclusion criteria: nine patients received a halothane-based anesthetic and six patients received a fentanyl-based anesthetic. When the data were analyzed by pooling both groups, the differences between pre- and postoperative sleep studies demonstrated a reduction in the number of obstructive events and less severe oxygen desaturations on the operative night. Total sleep time between the two sleep studies decreased from 371 +/- 13 to 304 +/- 14 mins. The number of obstructive apnea events/hr decreased as well. The lowest oxygen saturation measured during rapid eye movement sleep was 78 +/- 5% preoperatively and 92 +/- 1% postoperatively. CONCLUSIONS: Our data suggest that children without underlying medical conditions, neuromotor diseases, or carniofacial abnormalities, 1 to 18 yrs of age, who suffer from mild obstructive sleep apnea, have improvements documented by polysomnography on the night of surgery following adenotonsillectomy and do not necessarily need to be monitored intensively. These findings were not significantly affected by the choice of intraoperative anesthetic. PMID- 8706487 TI - Critical illness polyneuropathy: clinical findings and outcomes of a frequent cause of neuromuscular weaning failure. AB - OBJECTIVE: To describe clinical and electrophysiologic features and outcomes of critically ill patients with neuromuscular causes of failure to wean from mechanical ventilator support. DESIGN: A prospective, consecutive, case series. SETTING: Neurological, neurosurgical, and medical intensive care units in a university hospital. PATIENTS: Seven patients during a 3-yr period with failure to wean from mechanical ventilation not explained by pulmonary complications. INTERVENTIONS: Muscle and nerve biopsy in three patients. MEASUREMENTS AND MAIN RESULTS: Detailed electrodiagnostic studies were done in all patients 3 to 6 wks (median 4.5) after the onset of the acute illness and were repeated 3 months to 3.5 yrs later in those patients who survived. Primary illnesses included various intracranial and medical conditions. All patients had moderate-to-severe limb weakness with marked muscle atrophy. Tendon reflexes were decreased in three patients, exaggerated in two patients with intracranial lesions, and absent in two patients. Electromyography demonstrated severe acute denervation, with striking involvement of proximal muscles. Muscle and nerve biopsies showed severe neurogenic atrophy and axonal degeneration without inflammation. There was no evidence of primary myopathy. Two patients died of complications of sepsis. Of the survivors, three patients had no further weakness at the time of reexamination, except for peroneal nerve palsy in one patient. Two patients, still in the recovery period, showed markedly improved conditions but still showed slight weakness of the proximal muscles. By electrophysiology, signs of chronic neurogenic damage were demonstrable in all survivors at follow-up. CONCLUSIONS: Critical illness polyneuropathy is a frequent cause of neuromuscular weaning failure in critically ill patients, regardless of the type of primary illness. Involvement of proximal (including facial and paraspinal) muscles is striking. Tendon reflexes are often preserved. Patients with central nervous system injury may likewise develop critical illness polyneuropathy. In these latter patients, tendon reflexes may even be exaggerated. Recovery from critical illness polyneuropathy is usually rapid and clinically complete, although incomplete on electrodiagnostic study. Residual peripheral nerve lesion, generally of the peroneal nerve, is the most frequent feature of incomplete recovery. The need for careful electrophysiologic testing is emphasized to clarify the nature and extent of neuromuscular disturbances in critically ill patients. Failure to recognize the development of neuropathy in these patients may lead to erroneous conclusions about the ability to wean them from the ventilator. PMID- 8706488 TI - Cerebral oxygenation monitoring by near-infrared spectroscopy is not clinically useful in patients with severe closed-head injury: a comparison with jugular venous bulb oximetry. AB - OBJECTIVE: To compare continuous jugular venous bulb oximetry and cerebral near infrared spectroscopy in patients with severe closed head injury. DESIGN: A prospective observational study. SETTING: Intensive care unit of a major teaching hospital. PATIENTS: Adults (n = 10) with severe closed-head injury (Glasgow Coma Scale score of < or = 8). INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Jugular venous bulb oximetry, cerebral near-infrared spectroscopy, and cerebral perfusion pressure were measured continuously. A total of 3,691 paired measurements of near-infrared spectroscopy and jugular venous bulb oximetry were analyzed. Poor correlation (r2 = .04) between paired measurements and wide limits of agreement (-13% to +21%) were demonstrated. The mean difference between measurements was +/- 4% and the standard deviation of the mean difference was +/- 8.69%. The data were subsequently grouped according to three clinically significant subgroups of jugular venous bulb oxygen saturation reflecting low ( < 55%), normal (55% to 75%) and high ( > 75%) saturation values. Poor correlation and wide limits of agreement between the two methods of measurement were observed in all groups. Values recorded by near-infrared spectroscopy did not significantly change between the groups, and 14 clinically significant episodes of jugular venous bulb desaturation were not detected by near-infrared spectroscopy. CONCLUSIONS: Tissue oxygen saturation determined by near-infrared spectroscopy does not reflect significant changes in cerebral oxygenation detected by the global measurement of jugular venous bulb oximetry. This finding may be explained by inadequate signal detection and inaccuracies in the algorithm used to filter out extracranial components. Until these technical difficulties are addressed, near-infrared spectroscopy, as measured by the machine assessed in this study, cannot be routinely recommended for assessment of cerebral oxygenation in patients with acute head injury. PMID- 8706489 TI - Gastric intramucosal pH and multiple organ injury: impact of ischemia-reperfusion and xanthine oxidase. AB - OBJECTIVES: To determine if gastric intramucosal pH is affected by hepatoneteric ischemia-reperfusion. We additionally proposed to determine if changes in gastric mucosal hydrogen ion concentration are associated with liver and lung injury following hepatoenteric ischemia-reperfusion. Finally, we hypothesized that gastric intramucosal pH is influenced by xanthine oxidase, an oxidant-generating enzyme released after hepatoenteric ischemia-reperfusion. DESIGN: Randomized, controlled, animal study. SETTING: University-based animal research facility. SUBJECTS: Thirty-six New Zealand white male rabbits (2 to 3 kg). INTERVENTIONS: Anesthetized rabbits were randomly assigned to one of four groups (n = 9 per group): a) sham-operated group; b) sham-operated group pretreated with sodium tungstate (xanthine oxidase inactivator); c) aorta occlusion group; and d) aorta occlusion group pretreated with sodium tungstate. Descending thoracic aorta occlusion was maintained for 40 mins with a 4-Fr Fogarty embolectomy catheter, followed by 2 hrs of reperfusion. MEASUREMENTS AND MAIN RESULTS: Gastric tonometry was performed after completion of the surgical preparation (30-min equilibration) and at 30, 60, 90, and 120 mins of reperfusion. Plasma alanine aminotransferase activity was determined at 120 mins of reperfusion to assess hepatic injury. Bronchoalveolar lavage of the right lung was performed after 120 mins of reperfusion, and the protein content was determined as a measure of pulmonary alveolar-capillary membrane compromise. Descending thoracic aorta occlusion resulted in a significant decrease in gastric intramucosal pH as compared with sham-operated rabbits (p < .001). The change in gastric mucosal hydrogen ion concentration was significantly associated with plasma alanine aminotransferase activity (r2 = .48, p < .01) and bronchoalveolar protein content (r2 = .51, p < .01). Xanthine oxidase inactivation significantly improved gastric intramucosal pH after aortic occlusion and reperfusion (p < .001), with a concomitant attenuation of the release of plasma alanine aminotransferase (p < .05) and accumulation of bronchoalveolar protein (p < .05) during reperfusion. CONCLUSIONS: Gastric intramucosal pH was significantly decreased after hepatoenteric ischemia-reperfusion. Furthermore, an increase in gastric intramucosal hydrogen ion concentration was associated with a concomitant increase in tissue injury, a presumed harbinger of multiple organ failure. Gastric intramucosal pH values improved during reperfusion after xanthine oxidase inactivation, concomitant with attenuation of hepatic and pulmonary injury. Gastric tonometry is an important clinical tool that can provide critical insight into the pathogenesis of multiple organ injury after hepatoenteric ischemia reperfusion. Gastric tonometry may aid in the rapid assessment of pharmacologic interventions designed to attenuate multiple organ injury in similar clinical settings (e.g., trauma, shock, major vascular surgery). PMID- 8706490 TI - Endotoxic shock alters distribution of blood flow within the intestinal wall. AB - OBJECTIVE: To investigate whether a redistribution of blood flow from the mucosa to the muscular layer of the intestinal wall contributes to the observed increased arterial-mucosal Pco2 gradient and the decreased mucosal tonometric pH during endotoxic shock. DESIGN: A prospective, controlled, animal study. SETTING: Animal laboratory in a university medical center. SUBJECTS: Ten domestic pigs. INTERVENTIONS: Pigs were anesthetized with ketamine and pentobarbital, mechanically ventilated, hemodynamically monitored, and then challenged with Escherichia coli endotoxin (10 micrograms/ kg i.v.). MEASUREMENTS AND MAIN RESULTS: Cardiac output, mesenteric artery blood flow, and systemic, pulmonary, and portal pressures were measured. Intestinal mucosa tonometric Pco2 and pH were determined with saline-filled balloon tonometers. The tissue blood flow to the mucosa and the muscular layer were independently measured with colored microspheres, using the arterial reference sample method. Thus, total intestinal blood flow was evaluated with respect to its transmural (mucosa vs. muscularis) and geographical (proximal jejunum, mid-small intestine, and terminal ileum) distribution. Endotoxin administration with fluid resuscitation induced a distributive shock with a decrease in intestinal mucosa tonometric pH. Under endotoxemic conditions, the mucosal flow increased in each geographical area, with the increase being larger proximally in the jejunum than distally in the ileum. The mucosal tonometric pH was found to correlate inversely with mucosal blood flow. The increase in blood flow to the mucosa was balanced by a decrease in blood flow to the muscularis, with total mesenteric flow remaining unchanged. CONCLUSIONS: Mucosal hypoperfusion does not account for the acidotic mucosal tonometric pH in endotoxic shock. The results suggest either a primary cytotoxic effect or an enhanced counter-current-mediated hypoxic insult in the apical villus. The decrease in blood flow to the muscularis may contribute to loss of intestinal wall peristaltic activity and structural wall integrity. PMID- 8706491 TI - Hormonal and hemodynamic changes in a validated animal model of brain death. AB - OBJECTIVE: To examine the hormonal and hemodynamic changes in a validated animal model of brain death. DESIGN: Prospective, controlled study. SETTING: Experimental research laboratory. SUBJECTS: Adult male mongrel dogs (n = 10). INTERVENTIONS: Brain death was induced by inflation of a subdural balloon in ten mongrel dogs weighing 23 to 30 kg and validated neuropathologically. The hearts were instrumented with micromanometers and ultrasonic flow probes to measure cardiovascular changes. No inotropic or vasoactive support was given. Hemodynamic stability was maintained with intravenous fluids. Blood samples and hemodynamic readings were collected before and after the induction of brain death. MEASUREMENTS AND MAIN RESULTS: A Cushing reflex, followed by a hyperdynamic response and diabetes insipidus, occurred in every animal following brain death. Mean arterial pressure, heart rate, contractility, and cardiac output increased to > 350 mm Hg, 230 beats/min, 4200 mm Hg/sec, and 2.8 L/min, respectively, at the peak of this phenomenon before returning to baseline. A plasma catecholamine surge was observed in every animal 15 mins after brain death, while the circulating concentrations of the pituitary gland hormones vasopressin and adrenocorticotrophic hormone decreased significantly after 15 and 45 mins of brain death, respectively, and continued to decrease throughout the experiments. Circulating triiodothyronine, thyroxine, and glucagon concentrations decreased significantly (p < .01) from 0.58 +/- 0.05 ng/mL, 2.20 +/- 0.15 micrograms/dL, and 49.7 +/- 9.1 pg/mL, respectively, to 0.34 +/- 0.03 ng/mL, 1.14 +/- 1.14 micrograms/dL, and 6.9 +/- 1.4 pg/mL, respectively, 420 mins after brain death. The hematocrit increased significantly 15 mins after brain death and then gradually decreased throughout the duration of the experiments. CONCLUSIONS: In a validated animal model of brain death, significant decreases in the circulating concentrations of stress hormones, as well as hemodynamic instability, occurred after brain death. Measurements of plasma adrenocorticotrophic hormone and vasopressin values may be useful as diagnostic predictors of brain death. Furthermore, the observed changes may contribute to organ dysfunction after brain death and may necessitate hormonal as well as inotropic and vasoactive support to maintain donor organ function in the clinical setting. PMID- 8706492 TI - Effects of graded doses of vasopressin on median fibrillation frequency in a porcine model of cardiopulmonary resuscitation: results of a prospective, randomized, controlled trial. AB - OBJECTIVE: To assess the effects of graded doses of vasopressin vs. saline on median fibrillation frequency and defibrillation success in a porcine model of cardiopulmonary resuscitation. DESIGN: Prospective, randomized, controlled trial. SETTING: Animal laboratory in a university medical center. SUBJECTS: Twenty-eight domestic pigs (body weight between 26 and 31 kg), aged 12 to 14 wks. INTERVENTIONS AND MAIN RESULTS: After 4 mins of ventricular fibrillation and 3 mins of closed-chest cardiopulmonary resuscitation, the animals were allocated to receive either 0.2 U/kg of vasopressin (n = 7), 0.4 U/kg of vasopressin (n = 7), 0.8 U/kg of vasopressin (n = 7), or 10 mL of saline (n = 7, control group). Using radiolabeled microspheres, myocardial blood flow rates during cardiopulmonary resuscitation-before drug administration and 90 secs and 5 mins after drug administration-were as follows in the four groups (mean +/- SEM): 18.8 +/- 0.9, 17.2 +/- 1.1, and 14.6 +/- 1.4 mL/min/100 g in the control group; 17.8 +/- 2.2, 49.6 +/- 6.3 (p < .01 vs. control group), and 29.4 +/- 3.1 mL/min/100 g (p < .05 vs. control group) in the group receiving 0.2 U/kg of vasopressin; 17.1 +/- 1.0, 52.4 +/- 7.5 (p < .01 vs. control group), and 52.2 +/- 5.8 mL/min/100 g (p < .001 vs. control group) in the group receiving 0.4 U/kg of vasopressin; and 18.1 +/- 1.6, 94.9 +/- 9.2 (p < .001 vs. control group), and 57.2 +/- 6.3 mL/min/100 g (p < .001 vs. control group) in the group receiving 0.8 U/kg of vasopressin. Using spectral analysis, median frequencies of ventricular fibrillation-before drug administration and 90 secs and 5 mins after drug administration-were as follows in the four groups: 9.6 +/- 0.4, 8.5 +/- 0.8, and 7.2 +/- 1.0 Hz in the control group; 9.7 +/- 0.5, 12.9 +/- 0.8 (p < .01 vs. control group), and 12.7 +/- 0.8 Hz (p < .001 vs. control group) in the group receiving 0.2 U/kg of vasopressin; 10.3 +/- 0.2, 12.7 +/- 0.9 (p < .01 vs. control group), and 12.8 +/- 0.7 Hz (p < .001 vs. control group) in the group receiving 0.4 U/kg of vasopressin; and 10.0 +/- 0.9, 14.1 +/- 0.9 (p < .001 vs. control group), and 12.5 +/- 0.9 Hz (p < .001 vs. control group) in the group receiving 0.8 U/kg of vasopressin at the same points in time. Median frequency before the first defibrillation attempt was 12.3 +/- 0.4 Hz in the resuscitated animals (n = 19) and 8.2 +/- 1.2 Hz in the nonresuscitated animals (n = 9) (p < .001). CONCLUSIONS: This study contributes to the characterization of the effect of increasing global myocardial blood flow on median fibrillation frequency after administration of graded doses of vasopressin in a porcine model of ventricular fibrillation. Interventions such as vasopressor treatment that increase fibrillation frequency improve the chance of successful defibrillation. PMID- 8706493 TI - Effects of E-selectin and P-selectin blockade on neutrophil sequestration in tissues and neutrophil oxidative burst in burned rats. AB - OBJECTIVE: Neutrophil deposition in tissues (leukosequestration) after shock may produce local tissue injury from proteases and high-energy oxygen species released from sequestered neutrophils. The initial step in the binding of neutrophils to capillary endothelium is the interaction of adhesion molecule (selectin) receptors between neutrophils and endothelial cells. We quantified leukosequestration in the tissues of burned rats using two methods of analysis: a) measurement of lung myeloperoxidase; and b) measurement of radiolabeled neutrophils and erythrocytes deposited in multiple tissues. We then determined the ability of a selectin receptor blocking agent to affect neutrophil deposition in tissues after burn injury. DESIGN: Prospective, controlled, laboratory study. SETTING: University research laboratory. SUBJECTS: Male Wistar rats (200 to 300 g). INTERVENTIONS: After tracheostomy and venous cannulation, rats received 17% total body surface area full-thickness contact burns and were resuscitated with saline (20 mL i.p.). Experimental animals received 2 mg/kg body weight i.v. administration of a P- and E-selectin blocking monoclonal antibody, CY-1747, immediately after burn. Lung tissue neutrophils were estimated by measuring myeloperoxidase in lung tissue. Neutrophil retention in lung, liver, spleen, gut, skin, muscle, kidney, and brain tissues was determined by removing (preburn) and differentially radiolabeling neutrophils (111In) and erythrocytes (51Cr), reinfusing cells 4.5 hrs after burn, and measuring tissue radioactivity 30 mins later. Edema was estimated by measuring extravasated 125 I-labeled albumin in the various tissues. Peripheral blood neutrophils were analyzed for intracellular hydrogen peroxide content, utilizing a fluorescent dye that reacts with hydrogen peroxide, coupled with analysis of cell fluorescence by flow cytometry. MEASUREMENTS AND MAIN RESULTS: Myeloperoxidase concentration was increased in lungs 5 hrs after burn (p < .05), indicating neutrophil deposition. Radioisotope studies demonstrated significant (p < .05) leukosequestration into the lung, gut, kidney, skin, and brain tissues at 5 hrs after burn. Flow cytometry showed increased intracellular hydrogen peroxide content in peripheral blood neutrophils 5 hrs after burn. Tissue edema, manifested by radiolabeled albumin retention, was not seen in any tissues. Postburn neutrophil deposition in lungs and liver was blocked (p < .05) by administration of CY-1747 after burn, but maximal neutrophil hydrogen peroxide content was unaffected. CONCLUSION: Burn injury in rats results in accumulation of neutrophils in multiple tissues. Neutrophil deposition in the lungs and liver is blocked by administration of the E/P-selectin blocking antibody, CY-1747. Since sequestration of metabolically active neutrophils may induce tissue injury, therapies that block postburn leukosequestration may improve clinical outcomes by limiting remote tissue injury. PMID- 8706494 TI - Alterations of myocardial and vascular adrenergic receptor-mediated responses in Escherichia coli-induced septic shock in the rat. AB - OBJECTIVES: To investigate responsiveness to exogenous catecholamines in rat bacteremic shock by studying both myocardial and vascular functional parameters; to determine in the same study the relationship of these parameters with other relevant biological parameters of the adrenergic pathway, such as myocardial beta adrenergic receptors and cyclic adenosine monophosphate (cAMP); and to indirectly approach the roles of tumor necrosis factor-alpha (TNF-alpha) and nitric oxide. DESIGN: Experimental, comparative study. SETTING: Laboratory in a university hospital. SUBJECTS: Male Sprague-Dawley rats, weighing 270 to 320 g. INTERVENTIONS: Intravenous injection of live Escherichia coli DH5 alpha (2 x 10(10) organisms/kg) or saline (0.6 mL) and comparison of the two groups. MEASUREMENTS AND MAIN RESULTS: Mean arterial pressure and heart rate (HR) were recorded, and circulating TNF-alpha concentrations were measured, during the first 3 hrs after E. coli administration. Myocardial and vascular functional parameters were obtained, respectively, from Langendorff-perfused hearts and isolated aortic rings. Adrenergic biochemical parameters (catecholamines, density and affinity of beta-receptors, and isoproterenol-stimulated myocardial cAMP) were determined 3 hrs after E. coli injection. Mean arterial pressure decreased within 5 to 60 mins after bacteria injection and returned to basal levels in the last 2 hrs; HR was unchanged. Serum TNF-alpha concentrations peaked at 120 mins (7333 +/- 672 pg/mL) and were still increased at 3 hrs. Plasma concentrations of epinephrine and norepinephrine were significantly (p < .05) increased. Baseline values for differential left ventricular pressure and coronary flow were significantly (p < .0001, p < .001, respectively) reduced; HR remained unchanged. Isoproterenol induced a similar increase in differential left ventricular pressure and in HR. There was no decrease in the functional myocardial response to adrenergic stimulation. beta-adrenergic receptors were similar in density and in affinity in the two groups. Isoproterenol-stimulated myocardial cAMP was significantly (p < .01) reduced compared with the control group. In aortic rings, bacteria administration significantly (p < .01) shifted the dose-response curve to norepinephrine to the right, both in the presence and absence of endothelium. NG-monomethyl-L-arginine significantly increased the contractions to attain the control level: p < .001 in presence of endothelium; p < .05 in absence of endothelium. CONCLUSIONS: In ex vivo experiments, 3 hrs after E. coli injection, vascular responsiveness was sharply decreased. This impaired response was improved by inhibition of nitric oxide. The heart, nevertheless, was still able to modulate its inotropic and chronotropic response to isoproterenol, even though an impaired beta-adrenergic-receptor stimulation of cAMP was already present. PMID- 8706495 TI - Positive end-expiratory pressure-induced hemodynamic changes are reflected in the arterial pressure waveform. AB - OBJECTIVE: To examine whether the hemodynamic changes due to mechanical ventilation with positive end-expiratory pressure (PEEP) can be assessed by the respiratory-induced variations in the arterial pressure waveform during normovolemia and experimental acute ventricular failure. DESIGN: Prospective, controlled experimental study. SETTING: Institutional experimental laboratory. SUBJECTS: Adult mongrel dogs. INTERVENTIONS: Experimental acute ventricular failure was induced by the infusion of pentobarbital (a cardiodepressant) and methoxamine (a vasoconstrictor), combined with volume loading. Both the control and acute ventricular failure groups were subjected to ventilation with incremental levels of PEEP up to 20 cm H2O. MEASUREMENTS AND MAIN RESULTS: Cardiac function was evaluated by cardiac output and left and right ventricular change in pressure over time (dP/dt) measurements. Arterial pressure waveform analysis was performed by measuring the systolic pressure variation, which is the difference between the maximal and minimal systolic blood pressure values during one mechanical breath. The components of the systolic pressure variation, namely, dUp and dDown, which are the increase and decrease in the systolic pressure during the mechanical breath relative to the systolic pressure during apnea, were also measured at each PEEP level. PEEP caused significant reduction of cardiac output in normovolemic dogs, and was associated with significant increases in systolic pressure variation and dDown. Acute ventricular failure decreased the variations in the systolic pressure and caused the dDown component to disappear. The application of PEEP did not affect cardiac output in dogs with acute ventricular failure, nor did it change systolic pressure variation and the dDown. CONCLUSIONS: Analysis of arterial pressure waveforms during mechanical ventilation reflected the decrease in cardiac output in dogs with normal cardiac function subjected to incremental PEEP. In dogs with acute ventricular failure in which PEEP did not affect cardiac output, the systolic pressure variation was similarly unaffected by PEEP. In the absence of cardiac output measurement during mechanical ventilation with PEEP, the analysis of the respiratory variations in the arterial pressure waveform may be useful in assessing changes in cardiac output. PMID- 8706496 TI - Effects of nitric oxide on hyperinflation-induced pulmonary hypertension in the isolated-perfused lung. AB - OBJECTIVE: To determine if nitric oxide decreases pulmonary vascular resistance in hyperinflation-induced pulmonary hypertension. DESIGN: Isolated-perfused lamb lung model. SETTING: Experimental animal laboratory in a university setting. SUBJECTS: Ten isolated-perfused lamb lungs harvested from subjects with a mean age of 29 days. INTERVENTIONS: After induction of anesthesia, endotracheal intubation, and mechanical ventilation, lungs were perfused via an extracorporeal circuit. Ventilatory pressures were set to provide tidal volumes of 10 mL/kg and ventilatory rates were adjusted to maintain a Paco2 of 40 +/- 5 torr (3.5 +/- 0.7 kPa). The perfusion system consisted of a blood reservoir, a membrane oxygenator, and a nonocclusive roller pump. Blood flow was increased progressively to 50 mL/kg/min, maintaining a pulmonary arterial pressure of < 25 mm Hg and a left atrial pressure between 2 and 5 mm Hg. End-expiratory lung volume was measured using a nitrogen washout method. Baseline data were collected after a 1-hr stabilization period. Lung volume was increased to achieve 25% (moderate hyperinflation) and 50% (severe hyperinflation) increments in pulmonary vascular resistance. Nitric oxide (80 parts per million) was administered to the preparation after each increment in lung volume. MEASUREMENTS AND MAIN RESULTS: Mean pulmonary arterial pressure, mean left atrial pressure, pulmonary vascular resistance, and static lung compliance were measured at baseline and after moderate and severe hyperinflation, both before and after nitric oxide administration. Significant decreases in pulmonary vascular resistance were found when the preparation was ventilated with nitric oxide at baseline (43% decrease) and during hyperinflation induced pulmonary hypertension at both moderate (31% decrease) and severe (23% decrease) levels of hyperinflation. CONCLUSIONS: Inhaled nitric oxide significantly reduces pulmonary vascular resistance, even when pulmonary hypertension is induced by airway hyperinflation and supraphysiologic lung volumes. These data suggest that the use of nitric oxide following lung transplantation may allow for effective management of pulmonary hypertension in patients who receive allografts from undersized donors. Further clinical experience will be crucial in precisely defining the range of donor recipient size mismatch that can be adequately managed and the time course over which nitric oxide can be administered safely and effectively to these patients. PMID- 8706497 TI - Predicting outcome in children with severe acute respiratory failure treated with high-frequency ventilation. AB - OBJECTIVES: a) To demonstrate the effect of high-frequency ventilation on gas exchange in children with severe acute respiratory failure unresponsive to conventional ventilation; b) to identify patients at high risk of death early after institution of high-frequency ventilation. SETTING: Tertiary care pediatric intensive care unit in a university hospital. DESIGN: A cross-sectional, observational study with factorial design. PATIENTS: Thirty-one patients with severe acute respiratory failure defined as a Pao2/F1o2 of < 150 torr (< 20 kPa) with a positive end-expiratory pressure of > or = 8 cm H2O and/or Paco2 of > 60 torr (> 8 kPa) with an arterial pH < 7.25. INTERVENTIONS: Patients received either high-frequency oscillation or jet ventilation if respiratory failure was unresponsive to conventional ventilation and if the underlying disease process was deemed reversible. MEASUREMENTS AND MAIN RESULTS: Thirty-one children were managed with high-frequency ventilation, 11 children with jet and 20 children with oscillator. Arterial blood gases and level of ventilatory support were recorded before and at 6, 24, 48, 72, and 96 hrs after institution of high frequency ventilation. There was an improvement in an arterial pH, Paco2, Pao2, and Pao2/FID2 6 hrs after institution of high-frequency ventilation (p < .01). This improvement, along with decreased need for oxygen, was sustained through the subsequent course. Twenty-three (74%) of 31 children treated with high-frequency ventilation survived. Survivors showed an increase in an arterial pH, Pao2, Pao2/FIO2, and a decrease in Paco2 within 6 hrs, whereas nonsurvivors did not. Oxygenation index was the best predictor of outcome. A combination of an initial oxygenation index of > 20 and failure to decrease the oxygenation index by > 20% by 6 hrs after initiation of high-frequency ventilation predicted death with 88% (7/8) sensitivity and 83% (19/23) specificity, with an odds ratio of 33 (p = .0036, 95% confidence interval 3-365). CONCLUSIONS: In patients with potentially reversible underlying diseases resulting in severe acute respiratory failure that is unresponsive to conventional ventilation, high-frequency ventilation improves gas exchange in a rapid and sustained fashion. The magnitude of impaired oxygenation and its improvement after high-frequency ventilation can predict outcome within 6 hrs. PMID- 8706498 TI - Comparative assessment of pediatric intensive care in Moscow, the Russian Federation: a prospective, multicenter study. AB - OBJECTIVE: Comparative assessment of pediatric intensive care. DESIGN: Prospective multicenter study. SETTING: Four pediatric intensive care units in Moscow, the Russian Federation. PATIENTS: Consecutive unselected admissions (n = 583), < or = 14 yrs of age, in a 6-month period. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Effectiveness was defined as the ratio of observed to predicted mortality, based on prediction by Pediatric Risk of Mortality (PRISM) severity of illness scoring. Efficiency (on the day of admission only) depended on either a mortality risk of > 1% or the administration of intensive care unit-dependent therapies. In all four hospitals, observed mortality rates were higher than expected, with a range of standardized mortality ratios between 1.10 and 1.83 (mean 1.32). The excess mortality was found in the low- and medium risk strata (risk of mortality of < 1% to 15%). Admission efficiency ratings did not fluctuate greatly between institutions (mean 60.4%, range 55.7 to 65.9). CONCLUSIONS: We provided a quantitative description and assessment of pediatric intensive care in Moscow. Moderate efficiency may reflect a low threshold for ICU admission due to poor nurse/patient ratios on the wards. Effectiveness in the low and medium-risk strata is below standard, as compared with a Western reference population. Excess mortality was concentrated in the low- and medium-risk strata, and can only partially be explained by the inclusion of co-morbidity. Future analysis should focus on specific treatment protocols, protocol adherence, and the determination of infectious and therapeutic complications. PMID- 8706499 TI - Sepsis and the systemic inflammatory response syndrome: neuromuscular manifestations. AB - OBJECTIVE: To describe the various conditions of peripheral nerve, neuromuscular junction, and muscle associated with the systemic inflammatory response syndrome (SIRS). DATA SOURCES: Publications in the scientific literature and personal observations during the last 15 yrs. DATA EXTRACTION: Computer search of the literature and review of patient records relating to polyneuropathy, neuromuscular transmission defects, and myopathies associated with sepsis, the septic syndrome, and SIRS. SYNTHESIS: SIRS is a new concept in which infection and trauma induce a systemic inflammatory response affecting the microcirculation to organs throughout the body. The nervous system is commonly affected in the forms of septic encephalopathy and critical illness polyneuropathy. Neuromuscular blocking agents and corticosteroids may have additional toxic effects on the neuromuscular system that are manifest as transient neuromuscular blockade, an axonal motor neuropathy, or a thick filament myopathy. Clinical examination in the critical care unit is often unreliable and electrophysiologic studies, at times accompanied by magnetic resonance imaging of the spinal cord, measurement of the circulating creatine phosphokinase concentration, and muscle biopsy, are often necessary to establish the diagnosis. Variants of critical illness polyneuropathy may occur outside the critical care unit. The precise mechanism of these neuromuscular conditions is not known, and further basic research is needed. CONCLUSIONS: A variety of neuromuscular conditions complicates SIRS. The identification of these conditions is important in patient management and in rendering a prognosis. PMID- 8706500 TI - A personal experience with SIRS and MODS. PMID- 8706501 TI - Unilateral lung hyperinflation and auto-positive end-expiratory pressure due to a ruptured right hemidiaphragm. PMID- 8706502 TI - Nitric oxide inhibition in the treatment of septic shock. PMID- 8706503 TI - Near-infrared spectroscopy and cerebral hemodynamics. PMID- 8706504 TI - Near-infrared spectroscopy and cerebral hemodynamics. PMID- 8706505 TI - Percutaneous revascularization in peripheral arterial disease. PMID- 8706506 TI - Gender differences in children's normative beliefs about aggression: how do I hurt thee? Let me count the ways. AB - This research was designed to assess whether children view relationally manipulative behaviors (i.e., those behaviors labeled relational aggression in past research) as "aggressive." 2 studies were conducted to evaluate whether children view relationally manipulative behaviors as associated with 2 defining features of aggression, anger (Study 1) and intent to harm (Study 2). Results of Study 1 (n = 459, 9-12-year-olds) indicated that relational aggression was the most frequently cited angry behavior for girls' interactions whereas physical aggression was the most frequently cited angry behavior for boys' interactions. Results of Study 2 (n = 60, 9-11-year-olds) demonstrated that relational aggression and verbal insults were the most frequently cited harmful behaviors for girls whereas physical aggression and verbal insults were the most frequently cited harmful behaviors for boys. These studies provide the first evidence that children view relationally manipulative acts (relational aggression) as angry, harmful behaviors (i.e., as "aggressive"). PMID- 8706507 TI - Aggressive boys' hostile perceptual and response biases: the role of attention and impulsivity. AB - The present study addressed whether (1) aggressive boys show hostile biases or general deficits in social perception, (2) aggressive boys' social perceptual difficulties also characterize isolate and isolate-aggressive children, (3) aggressive, isolate, and isolate-aggressive boys' social perceptual difficulties are attributable to inattention and impulsivity, and (4) aggressive and nonaggressive boys differ in the links between social perception and proposed behavioral responses. Aggressive boys demonstrated hostile biases, but not general deficits, in intention-cue detection relative to average-status boys. Isolate-aggressive boys resembled aggressive boys in social perception, whereas isolate boys showed mild deficits relative to average-status boys. Although isolates' general deficits were predominantly accounted for by inattention and impulsivity, aggressives' and isolate-aggressives' hostile biases remained after these problems were statistically controlled. The aggressive groups proposed aggressive responses much more frequently than the nonaggressive groups following intentions perceived as nonhostile. Measures corresponding to several stages of Dodge's social information processing model discriminated the aggressive from nonaggressive groups, thus providing support for this model. PMID- 8706508 TI - Behavior segmentation by boys as a function of aggressiveness and prior information. AB - Previous research documents a hostile attribution bias by aggressive boys when considering a social situation with ambiguous cues as to the intention of the participants. The present research evaluated whether this bias extends to the manner in which boys process incoming information. Aggressiveness of boys in grades 3-6 was assessed with peer behavior nominations. In a separate session, each child viewed a videotape of 2 boys playing tag on a playground and segmented the actions using a standard behavior segmentation procedure. A critical event occurred when one of the boys fell down after being tagged, slowly got up, then resumed the game. Prior to viewing the videotape each child received one of three information conditions: benign (the boys were friends), hostile (the boys did not like one another), or neutral (no specific information about the relationship between the boys). Aggressiveness predicted change in segmentation after the critical event only in the neutral prior information condition. Thus aggressiveness, found in other research to be associated with a hostile attribution bias, also was found to be related to differences in the organization of incoming information. PMID- 8706509 TI - A comparison of aggressive-rejected and nonaggressive-rejected children's interpretations of self-directed and other-directed rejection. AB - The hypothesis that aggressive-rejected children are unaware of their social status because they are self-protective when processing negative peer feedback was tested in 3 studies. In Study 1, fourth-grade girls and boys were asked to name peers they liked or disliked, as well as peers they thought liked or disliked them. Comparisons of aggressive-rejected, nonaggressive-rejected, and average status groups revealed that aggressive-rejected children were more unrealistic in their assessments of their social status than were nonaggressive rejected children. In Study 2, rejected and average boys identified in Study 1 were asked to name who they thought liked or disliked other children from their classroom. Comparisons of perceived and actual nominations for peers revealed that aggressive-rejected children were able to assess the social status of others as well as did nonaggressive-rejected and average status children. Because the difficulties aggressive-rejected children demonstrated in Study 1 did not generalize to judging the status of others in Study 2, the self-protective hypothesis was supported. Study 3 provided a parallel test of this hypothesis under more controlled conditions. Subjects from Study 2 viewed other children receiving rejection feedback from peers in videotaped interactions and received similar feedback themselves from experimental confederates. While all subjects rated self-directed feedback somewhat more positively than other-directed feedback, aggressive-rejected subjects had the largest self-favoring discrepancy between their judgments of self- and other-directed feedback. These findings also suggest that aggressive-rejected children may make self-protective "errors" when judging other children's negative feelings about them. Ethnicity differences in evaluating peer feedback emerged in Studies 1 and 3, raising questions about the impact of minority status on children's evaluations of rejection feedback. PMID- 8706510 TI - The Pictorial Scale of Perceived Competence and Social Acceptance: does it work with low-income urban children? AB - The present study represents a twofold investigation wherein (a) the construct validity of the Pictorial Scale of Perceived Competence and Social Acceptance for Young Children (PSPCSA) and (b) the developmental appropriateness of the PSPCSA were empirically examined for urban Head Start children. Data were collected from a sample of 476 African-American children (48-64 months) representing children enrolled in a large metropolitan Head Start program. In the first study, neither replication of the test author's statistical procedures nor a series of exploratory factor analyses produced psychologically meaningful constructs. A second study tested the assumption that the PSPCSA administration format is developmentally appropriate for preschool children. Results from this study did not support the assumption that the PSPCSA is developmentally appropriate for preschool children. Implications for establishing valid assessment methods for urban Head Start children were discussed. PMID- 8706511 TI - A multiperspective comparison of peer sociometric status groups in childhood and adolescence. AB - This study explores the sociometric status group differences in psychosocial adjustment and academic performance in various domains using multiple sources of information (teacher-, peer-, self-ratings, achievement data) and 2 age groups (elementary and secondary school students) in a different educational and cultural context. Gender differences in the profiles of the sociometric groups were also examined. The sample consisted of 1,041 elementary school (mean age = 11.4 years) and 862 secondary school (mean age = 14.3 years) students in public schools in Greece. Findings extended previous descriptions of rejected, neglected, and controversial groups based on the perceptions of all raters. Gender and age differences were found in the profiles of rejected and controversial groups, which were markedly distinguished from the other groups based on all data sets. Neglected children at both age levels were differentiated to a weaker degree. PMID- 8706512 TI - Friendship quality as a predictor of young children's early school adjustment. AB - The purpose of this study was to develop a method for assessing young children's perceptions of classroom friendships and to determine whether these perceptions were associated with their adjustment during the transition to grade school. Subscales tapping 5 friendship processes (e.g., validation, aid, disclosure of negative affect, exclusivity, conflict) were extracted from a 24-item friendship interview that was individually administered to 82 kindergarten children (M age = 5.61) who possessed a reciprocated and stable "best" friend in their classroom. Children's reports of the investigated friendship processes were found to be reliable, and processes such as perceived validation and conflict predicted children's satisfaction with their friendships, and the stability of these relationships. Perceived conflict in friendships was associated with multiple forms of school maladjustment for boys, including higher levels of school loneliness and avoidance and lower levels of school liking and engagement. For both boys and girls, validation and aid forecasted gains in perceived support from classmates, and aid also predicted improvements in children's school attitudes. Perceived exclusivity in friendships was associated with lower levels of achievement. These findings, and others reported in this article, are consistent with the hypothesis that the relational features of children's classroom friendships yield psychological benefits or costs (e.g., provisions) that, in turn, affect their development and adjustment. PMID- 8706513 TI - Cognitive and family-support mediators of preschool effectiveness: a confirmatory analysis. AB - Investigated in this study were the mediators of the effects of preschool intervention on children's school achievement in sixth grade. A confirmatory structural model developed in a previous study of third graders was tested with 360 low-income, mostly black children who were available at the 3-year follow-up. The model incorporated cognitive readiness at kindergarten entry and parent involvement in school (rated by teachers and parents) as primary mediators of preschool effectiveness. In sixth grade (age 12), preschool participation at ages 3 or 4 was significantly associated with higher reading achievement, higher math achievement, and with lower incidence of grade retention. Cognitive readiness and parent involvement in school significantly mediated the estimated effects of preschool participation on school achievement and grade retention 7 years postprogram. Teacher ratings of school adjustment, school mobility, and grade retention also contributed to the transmission of effects. This integrated model fit the data better than several alternative models, including those based on the cognitive-advantage and family-support hypotheses. PMID- 8706514 TI - Perceptions and attitudes of black students toward school, achievement, and other educational variables. AB - To provide data on the perceptions and attitudes of gifted Black early adolescents toward school, achievement, and other educational variables, the responses of gifted Black students (n = 48) were compared to Black students in a regular education (n = 50) and potentially gifted program (n = 50). Results indicate that gifted Black students held significantly different attitudes and perceptions than both potentially gifted students and regular education students relative to principles of the achievement ideology, attitudes toward school, attitudes toward academically successful students, and perceptions of parental achievement orientation. Of the three groups, gifted Black students were the most achievement oriented, and perceived strong parental achievement orientations; regular education students were the least optimistic and the most affected by negative peer relations and psychological issues. PMID- 8706515 TI - Adolescents' conceptions of teachers' authority and their relations to rule violations in school. AB - Conceptions of teachers' authority and reported misconduct regarding 20 moral, conventional, personal, contextually conventional, and prudential issues were assessed in 120 fifth, seventh, ninth, and eleventh graders (mean ages = 10.66, 12.88, 15.04, and 17.25 years, respectively). Adolescents viewed moral, conventional, and prudential issues as legitimately subject to teachers' authority and personal issues as under personal jurisdiction, but they were equivocal about contextually conventional issues. Fifth graders judged all acts as more legitimately subject to teachers' authority, all rule violations as more negative, and personal and prudential issues as personal more than did older students. Conventional misconduct was more frequent and moral misconduct was less frequent than other rule violations, but both were greater among boys than girls. Adolescents' negative rule evaluations, fewer rules, greater dislike for school, poorer grades, and living in single- or step-parent families predicted teacher- and self-reported misconduct. Relations to previous research on conceptions of adult authority, school misconduct, and autonomy development are discussed. PMID- 8706516 TI - Anxiety and the processing of emotionally threatening stimuli: distinctive patterns of selective attention among high- and low-test-anxious children. AB - There is substantial evidence that clinically referred and nonreferred high anxious adults selectively shift attention toward threatening stimuli. In contrast, low-anxious adults shift attention away from threatening stimuli. Recent evidence suggests that clinically referred anxious children also selectively attend to threatening information. The present study tested for the presence of such a bias in a nonreferred sample of high-anxious children and also included the first adequate test for an attentional bias away from threat among low-anxious children. 20 high- and 20 low-test-anxious children, 11-14 years of age, completed a task in which visual attention was indexed by latency to detect probes following emotionally threatening and neutral words. Results supported the predicted attentional bias toward threat cues among high-test-anxious children. Unexpectedly, the predicted attentional bias away from threat cues was found only among low-test-anxious boys. Low-test-anxious girls attended equally to threatening and neutral words. In sum, selective attention mechanisms influence children's processing of threatening information and may play a role in the regulation and dysregulation of childhood anxiety. PMID- 8706517 TI - Maternal attitudes and knowledge of child-rearing: associations with family and child outcomes. AB - A prospective, longitudinal study of a low birthweight, preterm cohort examined the effects of maternal knowledge of child development and concepts of child rearing on the quality of home environment and on child cognitive and behavioral outcomes. Measures of maternal knowledge at 12 months were found to be significantly associated with the quality of the home environment, the number of child behavior problems, and to a small but significant extent child Stanford Binet IQ at 36 months. Maternal characteristics were associated with both maternal knowledge and maternal behavior. Child characteristics, including birthweight, were not associated with maternal knowledge or concepts of development for most of the cohort. Subgroup analyses by race/ethnicity revealed a similar pattern of results. However, a measure of neonatal health status was shown to be significantly associated with cognitive outcome in the African American subgroup at 24 and 36 months. PMID- 8706518 TI - Multifaceted impact of self-efficacy beliefs on academic functioning. AB - This research analyzed the network of psychosocial influences through which efficacy beliefs affect academic achievement. Parents' sense of academic efficacy and aspirations for their children were linked to their children's scholastic achievement through their perceived academic capabilities and aspirations. Children's beliefs in their efficacy to regulate their own learning and academic attainments, in turn, contributed to scholastic achievement both independently and by promoting high academic aspirations and prosocial behavior and reducing vulnerability to feelings of futility and depression. Children's perceived social efficacy and efficacy to manage peer pressure for detrimental conduct also contributed to academic attainments but through partially different paths of affective and self-regulatory influence. The impact of perceived social efficacy was mediated through academic aspirations and a low level of depression. Perceived self-regulatory efficacy was related to academic achievement both directly and through adherence to moral self-sanctions for detrimental conduct and problem behavior that can subvert academic pursuits. Familial socioeconomic status was linked to children's academic achievement only indirectly through its effects on parental aspirations and children's prosocialness. The full set of self-efficacy, aspirational, and psychosocial factors accounted for a sizable share of the variance in academic achievement. PMID- 8706519 TI - Social class effects on northeastern Brazilian children's conceptions of areas of personal choice and social regulation. AB - 2 studies examined middle- and lower-class Brazilian children's concepts of personal choice and social regulation. In Study 1, interviews of 40 middle- and lower-class children (9 and 15 years old) revealed that children across classes distinguished moral from conventional issues on the bases of rule contingency and act generalizability criteria. Lower-class children, however, were less likely to view conventions as rule contingent and more likely to generalize conventional acts. In Study 2, interviews of 240 middle- and lower-class children (ages 8, 12, 16 years) found that across classes, children distinguished prudential issues from matters they treated as personal. Prudential issues were seen as subject to parental authority. Middle-class children were more likely to treat personal issues as matters of choice. With age, lower-class children increasingly tended to treat personal items as matters of choice, and by adolescence there were no class differences. Findings show that Brazilian children maintain a heterogeneous orientation to rules and authority which includes a domain of personal choice. Class differences indicate that hierarchical social structures affect children's sense of autonomy. However, developmental effects indicate that a domain of personal choice emerges among children across social classes. PMID- 8706521 TI - Context and development in family problem solving with preadolescent children. AB - This study analyzed correlates of social problem solving in 63 2-parent families with a preadolescent child during structured discussions in the home. The longitudinal design assessed families when the preadolescent was about 9.5 years old and again 2 years later. Links between problem solving and features of the immediate ecological context were examined. There was a substantial decline in the quality of family problem solving during preadolescence associated with changes in the participation and affective behavior of fathers and preadolescents. Although problem solving was not less effective on autonomy topics, it was markedly worse when parents, rather than the preadolescent, selected the topic for discussion. Preadolescent topic selection triggered a pattern of productive participation and affective behavior in both parents and preadolescents. The results clarify the nature of ambivalent parent-child relations just prior to puberty by specifying changes in family problem solving which correspond with preadolescent autonomy striving. PMID- 8706520 TI - The effects of work intensity on adolescent mental health, achievement, and behavioral adjustment: new evidence from a prospective study. AB - This article examines the effects of work intensity on adolescent mental health, academic achievement, and behavioral adjustment. Questionnaire data were collected yearly from an initial panel of 1,000 randomly selected ninth graders (14-15 years old). Consistent with other studies, students who worked at higher intensity engaged in more alcohol use. The methodological strengths of this research (a representative panel studied prospectively over a 4-year period with minimal attrition and an analysis incorporating key control and lagged variables) provide strong evidence that adolescent work fosters alcohol use. The contention that work of high intensity has deleterious effects on mental health, academic achievement, and 2 other indicators of behavioral adjustment did not withstand our stringent tests. However, high school seniors who worked at moderate intensity (1-20 hours per week) had higher grades than both nonworkers and students who worked more hours per week. PMID- 8706522 TI - Parent-child relationships, family problems-solving behavior, and sibling relationship quality: the moderating role of sibling temperaments. AB - In this study we sought to determine whether sibling temperaments moderated the associations of parent-child relationship quality and family problem-solving behavior with sibling relationship quality. Observational assessments of mother child and father-child relationship quality, family problem-solving behavior, and sibling relationship quality were obtained from the families of 49 pairs of brothers and 46 pairs of sisters. Mean ages were 10-2 for older siblings and 7-6 for younger siblings. Parent-reported child temperament assessments were also obtained. The links among mother-older child relationship quality, father-older child relationship quality, and sibling relationship quality were moderated by the older sibling's temperament. The younger and older siblings' temperaments moderated the associations between the quality of the father-younger sibling and sibling relationships. The association between family problem-solving behavior and sibling relationship quality was not moderated by sibling temperaments. PMID- 8706523 TI - The development of hierarchical representation of two-dimensional space. AB - Adults represent the location of a point in a 2-dimensional space using 2 independent dimensions. They encode location along these dimensions both at a fine-grained level and categorically. In reporting location, they combine and weight the fine-grained and categorical information. In Experiment 1, we found that children as young as 5 years use the same 2 independent dimensions in fine grained spatial coding of location in a circle as are used by adults-radius and angle. However, categorical coding and hierarchical combination are seen only for radius, at both 5 and 7 years. The adult pattern, where angle as well as radius is coded hierarchically, emerges by 9 years. Experiment 2 shows that there is nothing intrinsically difficult about the categorical coding of angular information; when angle is the only dimension to be encoded, younger children use hierarchical coding. Changes in 2-dimensional hierarchical coding may be due to cognitive load factors and to changes in ability to assign frames of reference. PMID- 8706524 TI - The dynamics of preschoolers' categorization choices. AB - The present research explored the effects of stimulus and task factors on preschoolers' (Experiments 1 and 3) and adults' (Experiment 2) tendency to categorize according to taxonomic relations, when those relations conflict with appearances. In Experiment 1, we examined the effects of and interactions among (a) available information, operationalized by using more- or less-informative stimulus types (objects vs. line drawings) and by the presence or absence of labeling, and (b) task constraints, operationalized by comparing sorting questions with inductive inferences questions. When provided with information that constrained the categorization decision, either through the availability of labels or a combination of enhanced physical informativeness of objects and an inference question, preschoolers reliably based their categorization decisions on taxonomic relations between physically dissimilar items. In Experiment 2, stimulus type (objects vs. line drawings) was shown to have a similar effect on adults. In Experiment 3, we examined the effects of stimulus type on preschoolers' inductive inferences and accuracy of naming. The effects in the two tasks were closely related, suggesting that the amount of available information affects different responses in similar ways. These data demonstrate the interactive effects of available information and task constraints on categorization decisions. PMID- 8706525 TI - Young children's understanding of thought bubbles and of thoughts. AB - In a series of 4 studies, we explored preschoolers' understanding of thought bubbles. Very few 3-year-olds or 4-year-olds we tested knew what a thought-bubble depiction was without instruction. But, if simply told that the thought bubble "shows what someone is thinking," the vast majority of 3-year-olds and 4-year olds easily understood the devices as depicting thoughts generally and individual thought contents specifically. In total, these children used thought-bubble depictions to ascertain the contents of characters' thoughts in a variety of situations; appropriately distinguished such depictions from mere associated actions or objects; described thought bubbles in the language of mental states; judged that persons' thoughts in these depictions were subjective in the sense of person-specific (and hence 2 people can have different thoughts about the same state of affairs); and judged that thought-bubble thoughts (a) were representational in the sense of depicting or showing some other state of affairs, (b) were mental and thus showed intangible, private, internal thoughts unlike real pictures or photographs, and (c) can be false, that is, can depict a person's misrepresentation of some state of affairs. We discuss the implications of these findings for young children's understanding of thoughts and thought bubbles, for their learning and comprehension of pictorial conventions, and for the use of thought bubbles to assess children's early understanding of mind. PMID- 8706526 TI - Continuities in emotion understanding from three to six years. AB - As part of a longitudinal study, 47 children who were tested on their understanding of basic emotions when they were 3 years old were tested again at 6 on their understanding of conflicting emotions. Significant stability in individual differences was found over this 3-year period. Antecedents to emotion understanding at 3 continued to be significantly related to children's understanding at 6, including participation in discourse about causality, positive interaction with older siblings, and language ability. Girls outperformed boys, and there was a suggestion that the girls' understanding of emotions was more closely associated than the boys' with the quality of their sibling relationships. Children's concurrent reports of negative experiences at home and at school were related to their sensitivity to the experience of ambivalent emotions. The results are discussed with respect to the social origins and implications of this core aspect of children's social-cognitive development. PMID- 8706527 TI - There's more to the picture than meets the eye: young children's difficulty understanding biased interpretation. AB - 5 experiments investigated children's understanding that expectations based on prior experience may influence a person's interpretation of ambiguous visual information. In Experiment 1, 4- and 5-year-olds were asked to infer a puppet's interpretation of a small, ambiguous portion of a line drawing after the puppet had been led to have an erroneous expectation about the drawing's identity. Children of both ages failed to ascribe to the puppet an interpretation consistent with the puppet's expectation. Instead, children attributed complete knowledge of the drawing to the puppet. In Experiment 2, the task was modified to reduce memory demands, but 4- and 5-year-olds continued to overlook the puppet's prior expectations when asked to infer the puppet's interpretation of an ambiguous scene. 6-year-olds responded correctly. In Experiment 3, 4- and 5-year olds correctly reported that an observer who saw a restricted view would not know what was in the drawing, but children did not realize that the observer's interpretation might be mistaken. Experiments 4 and 5 explored the possibility that children's errors reflect difficulty inhibiting their own knowledge when responding. The results are taken as evidence that understanding of interpretation begins at approximately age 6 years. PMID- 8706528 TI - Gender-schema development and children's constructive story memory: evidence for a developmental model. AB - The objectives of the present research were to (a) provide a developmental model based on script research for describing how changes in memory for gender-related information are related to changes in gender-role stereotypes, (b) examine developmental differences in the effect of stereotype manipulations on the construction of new memories, and (c) examine the relation between stereotyped activity preferences and memory for gender-related information. 4-, 6-, and 8 year-olds listened to a story in which characters performed behaviors typical and atypical of gender-role stereotypes. An introduction preceded the story in which story characters' activities and preferences were described as either consistent or inconsistent with gender-role stereotypes. Dependent variables were the percentage of typical and atypical story items correctly recognized and the percentage of false alarms made for new items. Gender-role knowledge, stereotyped preferences, and gender-role flexibility were assessed. Results for false alarms, but not hit rates, supported the hypotheses of the model: (a) 6-year-olds falsely recognized typical distractors more than atypical distractors (this effect was nonsignificant for 4- and 8-year-olds), (b) false alarms for atypical distractors decreased between ages 4 and 6, and (c) false alarms for typical distractors decreased between ages 6 and 8. Contrary to expectation, stereotype manipulation effects did not interact with age, but were influenced by gender. Stereotyped preferences were strongly related to memory for gender-related information for both males and females. Results are discussed in terms of developmental and individual differences in gender-schema strength and composition. PMID- 8706530 TI - Specifying the relation between novel and known: input affects the acquisition of novel color terms. AB - 4 studies investigate how differential input affects preschoolers' abilities to learn novel color words. 3-, 4-, and 5-year-old children saw objects in novel shapes and colors and heard a novel color label for the object. Labels were presented through ostensive definition (e.g., "See, it's mauve"), corrective linguistic contrast (e.g., "See, it's not purple; it's mauve"), or an inclusion statement (e.g., "See, it's mauve; it's a kind of purple"). 4- and 5-year-old children interpreted the novel word as a shape term when ostensive information was provided but as a color term when additional information, either contrastive or inclusive, specified a relation between the novel term and a known label for that color. Furthermore, children who consistently interpreted the novel word as a color word tended to treat the novel and known labels as mutually exclusive color terms if they heard contrastive information, whereas they tended to treat the words as hierarchically related if they heard inclusion information. 3-year olds generally did not make use of either type of information in determining the semantic domain of the novel word or the relation between terms. PMID- 8706529 TI - Why talk about mental states? The significance of children's conversations with friends, siblings, and mothers. AB - Natural language data from 38 47-month-olds recorded at home in unstructured observations were analyzed and comparisons made of characteristics of mental state term use in child-friend, child-sibling, and child-mother dyads. Significantly more references to mental states were made by the children in conversations with siblings and friends than with mothers. Frequent use of mental state terms by both partners was related to cooperative interaction in both child friend and child-sibling dyads and several associations were found with measures of language fluency, gender, and maternal education, although these varied across the 2 dyads. Children's use of mental state terms in conversations with siblings and friends was correlated with their performance on two false belief measures. Results highlight the importance of extending investigations into the social implications of the development of children's "theories of mind." PMID- 8706531 TI - The association between language and symbolic play at two years: evidence from deaf toddlers. AB - The association between expressive language and symbolic play was investigated in 3 groups of 2-year-olds: deaf children with hearing parents (dH), deaf children with deaf parents (dD), and hearing children with hearing parents (hH). (Each group included 6 girls and 4 boys.) 3 language-level groups were defined. The highest group was well into the vocabulary "explosion" and frequently produced multiword/sign utterances; a middle group was beginning the period of vocabulary expansion and occasionally produced utterances of more than 1 word/sign; a third group produced single word/sign utterances only, and had a limited vocabulary. Hearing status was associated with duration of symbolic play (deaf > hearing). Higher language levels were associated with more canonically sequenced and preplanned play, even when language delays were due to exogenous factors. PMID- 8706532 TI - Dampening of adrenocortical responses during infancy: normative changes and individual differences. AB - Changes in cortisol and behavioral responses were examined longitudinally in 83 infants (39 girls, 44 boys) tested at their well-baby exams with inoculations at 2, 4, 6, and 15 months (72 infants completed all testing). Another sample of 2-, 4-, and 6-month-olds (n = 18 per age) received mock exams without inoculations to determine early developmental changes to the exam procedures. Behavioral distress was coded every 30 sec during the exam, a 5-min inoculation period, and a 20-min recovery period. Salivary cortisol was obtained upon arrival at the clinic and 25 min after the beginning of the inoculation period. To the exam-inoculation procedures, cortisol responses were high at 2 months, decreased significantly between 2 and 4 months, remained comparable between 4 and 6 months, and then declined again between 6 and 15 months. Between 6 and 15 months there was also a decrease in pretest cortisol. By 15 months, significant increases in cortisol from pre- to posttest were no longer observed for most infants. To the exam-only procedures, cortisol responses decreased between 2 and 4 months, and by 4 months most infants failed to show pre- to posttest increases in cortisol. Behavioral distress decreased between 2 and 6 months, but increased again at 15 months. While crying and cortisol were modestly correlated during the 2- to 6-month exam inoculation procedures, at 15 months no significant correlations were obtained. Behavioral and hormonal reactions thus followed different ontogenetic paths and may provide different information about infant functioning. There was some evidence that the emergence of the circadian rhythm in cortisol might be related to the early decrease in cortisol response. PMID- 8706533 TI - The development of perceptual completion abilities: infants' perception of stationary, partially occluded objects. AB - We explored infants' ability to perceive stationary, partially occluded objects as connected units (Experiments 1 and 2) with specific appearances (Experiment 3). In each experiment, the infants saw 2 test events involving what appeared to adults to be a tall rectangular object whose middle portion was hidden behind a narrow screen. During the test events, the screen alternately uncovered and covered the object. In Experiments 1 and 2, removal of the screen revealed either a single, connected rectangle (complete object event) or an interrupted rectangle with a gap where the screen had been (broken object event). In Experiment 3, removal of the screen revealed either a rectangle (rectangle event) or a cross shaped object (cross-shape event). The pattern of infants' looking times at these events suggest that they perceive the unity of the partially occluded object by 6.5 months of age but did not perceive the form of the hidden part of the object until 8 months. The results of baseline control conditions support this interpretation. PMID- 8706534 TI - Infant affective reactions to the resumption of maternal interaction after the still-face. AB - To investigate infants' reactions to the Face-to-Face Still-Face Paradigm and in particular the reunion episode, 50 6-month-olds' affective, behavioral, and physiologic reactions were recorded and analyzed. Infants reacted to the still face with negative affect, a drop in vagal tone, and an increase in heart rate. By contrast, they reacted to the reunion episode with a mixed pattern of positive and negative affect. There was a carryover of negative affect from the still face, an increase in fussiness and crying, and a rebound of positive affect. During this episode, the infants' heart rate and vagal tone returned to initial levels. The data indicate that infant affective displays are specifically related to different interactive events, but that their physiologic reactions do not show the same level of specificity. The findings also highlight the complexity of the affective and reparatory processes that take place in mother-infant interactions. PMID- 8706535 TI - Expressions of the attachment relationship outside of the strange situation. AB - 41 preterm infants and 38 full-term infants and their mothers were observed at home at 8 and 12 months of age and in the Strange Situation at 18 months in order to compare expressions of attachment relationships in these 2 settings. There was 84% concordance in the distinctions between secure and nonsecure classifications of the mother-infant relationship made at home at 12 months and in the Strange Situation. Classifications of avoidant relationships also displayed high concordance, but only 6 of the 15 dyads classified at home as ambivalent were classified in the same way in the Strange Situation. Mothers in secure relationships as assessed in the Strange Situation were rated as more sensitive at both 8 and 12 months than mothers in either avoidant or ambivalent relationships, whereas the sensitivity of mothers in these two nonsecure relationships did not differ significantly. Infants in secure relationships in the Strange Situation were characterized by more effective secure base behavior and more affective sharing and enjoyment of physical contact, and they were less fussy or difficult during the 12-month home observations. PMID- 8706536 TI - Emotion regulation in two-year-olds: strategies and emotional expression in four contexts. AB - A descriptive study of the modulation of negative emotion in the toddler period was conducted by examining expressions of negative emotion and the strategies used to reduce or change these expressions. 6 strategies were identified and evaluated in terms of their frequency of use in different situations, relations with emotional expressiveness, and cross-situational consistency. 37 2-year-olds were seen in 2 laboratory contexts (delay and separation) each with 2 variants. Emotion regulation strategies and emotional expressiveness were coded from videotapes of children's behavior in these 4 situations. Findings suggest that active engagement was most commonly used and most negatively associated with child distress. Use of strategies varied by context, and there was more cross situational consistency in use of strategies that were more negatively or positively associated with distress within a given context than in use of particular strategies without consideration of their within-context significance. PMID- 8706537 TI - Getting angry at peers: associations with linking of the provocateur. AB - We examined how preschoolers coped with anger in interactions with well liked and not well liked peers. The free-play interactions of preschool-aged children (M age = 66.27 months) were observed for 6 months. The frequency, causes, and intensity of children's anger, as well as their anger-related reactions, were compared for incidents provoked by peers who were "really liked" to those provoked by peers who were liked only " a little bit." Although there were no differences in the intensity of anger provocations by well liked and not well liked provocateurs, children's responses to provocations by well liked peers were more controlled than was the case for anger provoked by peers who were not well liked. In general, boys were more responsive to how they felt about the provocateur than were girls. These findings suggest that anger episodes with well liked children were less stressful than those with peers who were not well liked. PMID- 8706538 TI - Display rules for anger, sadness, and pain: it depends on who is watching. AB - This study examined factors that may influence children's decisions to control or express their emotions including type of emotion (anger, sadness, physical pain), type of audience (mother, father, peer, alone), age, and sex. Children's reported use of display rules, reasons for their decisions, and reported method of expression were examined. Subjects were 32 boys and 32 girls in each of the first (M = 7.25 years old), third (M = 9.33 years old), and fifth grades (M = 11.75 years old). Regardless of the type of emotion experienced, children reported controlling their expression of emotion significantly more in the presence of peers than when they were with either their mother or father or when they were alone. Younger children reported expressing sadness and anger significantly more often than did older children, and girls were more likely than boys to report expressing sadness and pain. Children's primary reason for controlling their emotional expressions was the expectation of a negative interpersonal interaction following disclosure. PMID- 8706539 TI - The relations of children's dispositional prosocial behavior to emotionality, regulation, and social functioning. AB - The purpose of this study was to examine the relations of a measure of children's dispositional prosocial behavior (i.e., peer nominations) to individual differences in children's negative emotionality, regulation, and social functioning. Children with prosocial reputations tended to be high in constructive social skills (i.e., socially appropriate behavior and constructive coping) and attentional regulation, and low in negative emotionality. The relations of children's negative emotionality to prosocial reputation were moderated by level of dispositional attentional regulation. In addition, the relations of prosocial reputation to constructive social skills and parent reported negative emotionality (for girls) increased with age. Vagal tone, a marker of physiological regulation, was negatively related to girls' prosocial reputation. PMID- 8706540 TI - Social information-processing mechanisms in reactive and proactive aggression. AB - Theories of aggressive behavior and ethological observations in animals and children suggest the existence of distinct forms of reactive (hostile) and proactive (instrumental) aggression. Toward the validation of this distinction, groups of reactive aggressive, proactive aggressive, and nonaggressive children were identified (n = 624 9-12-year-olds). Social information-processing patterns were assessed in these groups by presenting hypothetical vignettes to subjects. 3 hypotheses were tested: (1) only the reactive-aggressive children would demonstrate hostile biases in their attributions of peers' intentions in provocation situations (because such biases are known to lead to reactive anger); (2) only proactive-aggressive children would evaluate aggression and its consequences in relatively positive ways (because proactive aggression is motivated by its expected external outcomes); and (3) proactive-aggressive children would select instrumental social goals rather than relational goals more often than nonaggressive children. All 3 hypotheses were at least partially supported. PMID- 8706541 TI - Total knee arthroplasty after proximal tibial osteotomy. AB - The cases of ten total knee arthroplasty after a proximal tibial osteotomy were compared with those of 50 primary total knee arthroplasty for osteoarthritis. The groups were matched according to age of patients, length of follow-up and type of prosthesis used. On the basis of HSS, an excellent or good result was obtained in 90% of patients who had had arthroplasty after osteotomy and in 94% of patients after a primary arthroplasty. Though some intraoperative difficulty may be encountered in a secondary prosthesis, osteotomy does not compromise a subsequent arthroplasty. PMID- 8706542 TI - Acetabular rim pathology secondary to congenital hip dysplasia in the adult. A radiographic study. AB - Major rim pathology is a precursor of osteoarthritis of the hip secondary to residual acetabular dysplasia. The symptoms are acute pain in the groin and impaired function; the anatomopathologic lesions consist in avulsions of the labrum from the bony rim, and separated bone fragments or "Os acetabuli", as well. A detailed radiographic, comparative inquiry to assess the preoperative morphology of 178 dysplastic hips which underwent a multiplanar periacetabular osteotomy was undertaken. The study showed that the 37 hips with a labral avulsion had a less pronounced anterior and lateral insufficiency of the acetabulum and a less pronounced lateral subluxation than dysplastic hips without these lesions. No specific radiologic features could be found in the 23 hips with bony fragments of the acetabular rim. The instability of the joint exerts abnormal stress on the acetabular rim which tends to tear it. This condition leads to rapid arthrotic degeneration of the hip; early diagnosis increases our ability to preserve the integrity of the joint through reliable reconstructive surgery. PMID- 8706543 TI - Changes in growth of the ilium after sopraacetabular osteotomy. Long-term evaluation. AB - It is commonly known that in acetabuloplasty according to the Pemberton-Zanoli method the insertion of the graft produces an immediate increase in the height of the iliac wing which in successive months could become hypergrowth caused by the hyperactivity of the underlying Y cartilage. A total of 40 cases of insufficiency of the acetabular roof treated by this method were examined based on measurements on follow-up radiograms and an evaluation of the spino-malleolar distance until puberty. The clinical evaluation showed that in 82.5% of cases there is dysmetria that is less than or equal to 0.5 cm. An evaluation of radiograms at the end of growth of the ilium [correction of ileum] did not reveal significant variations between the hemipelvis submitted to surgery and the contralateral one either in cases treated by acetabuloplasty alone or in those in which intertrochanteric osteotomy was associated. Acetabuloplasty according to the Pemberton-Zanoli method is thus effective in the treatment of acetabular dysplasia at a young age, obtaining long-lasting effects and ones that do not cause dysmetria in the adult. PMID- 8706544 TI - Recurrent posterior instability of the shoulder: surgical treatment. AB - A total of 11 cases of posterior instability of the shoulder surgically treated by simple capsuloplasty or associated with reinforcement of the posterior wall by bone graft or muscle transposition were reviewed. Accurate selection and preoperative evaluation of the patient when choosing surgery is very important. For surgery to succeed prolonged postoperative immobilization and an intense rehabilitative program to be initiated prior to surgery are necessary. Generalized ligamentous hyperlaxity constitutes a negative factor in prognosis, so that it is important to associate capsuloplasty with posterior bone block, in an attempt to avoid the risk of recurrence as further surgery would have an unfavorable outcome. PMID- 8706545 TI - Revision of the femoral prosthetic component according to the Wagner technique. AB - The authors report the preliminary results they obtained using the Wagner technique for revision of the prosthetic stem by transfemoral access. Between January 1992 and June 1993 a total of 20 patients were treated by reimplantation of the prosthetic stem according to the Wagner technique. At minimum 12-month follow-up, based on the Harris Score System, results were excellent, good and fair in 16 cases. Results were not satisfactory in 4 cases after sinking of the prosthesis exceeding 2 cm or due to mid-distal pain with loading. PMID- 8706546 TI - Lesions of the sciatic nerve in fractures of the acetabulum. AB - Between 1982 and 1992 a total of 211 fractures of the acetabulum were operated. There was lesion of the sciatic nerve in 21 cases, the pathogenesis and evolution of which were studied. Clinical follow-up was obtained at least 2 years after occurrence of neurological deficit and supported in 13 cases by electromyography. Based on an analysis of the literature and of these series it is observed how spontaneous recovery of paralysis is not rare, even if it requires a long period of time. The presence of various levels of lesion, which are not easily diagnosed, and which are not easily attacked surgically, calls for a prudent attitude with regard to neurolysis. PMID- 8706547 TI - Evaluation of several immunological parameters in patients with aseptic loosening of hip arthroplasty. AB - The authors evaluated several immunological parameters in patients with loosened hip arthroplasty in order to determine the state of sensitization with regard to the metallic constituents of the prosthesis. The results obtained by in vivo epicutaneous testing did not reveal a correlation between loosening and sensitization, as the patch test was positive in only 1 out of 16 cases examined. An evaluation of the lymphocyte subpopulation on peripheral blood demonstrated that patients with prostheses in Cr, Co, Mo, Ni alloy present significant lymphopenia, with a reduction in subpopulations CD4 and CD8 and a decrease in in vitro cytotoxic activity. The meaning of these modifications could be interpreted to be either a toxic effect products released from the implant, or as the recruitment of lymphocytes in the site of loosening, due to lymphocyte sequestration phenomena consequent to a cell-mediated hypersensitivity reaction. PMID- 8706548 TI - The effects of orthopaedic cements on osteoblastic cells cultured in vitro. AB - The authors have examined the response of osteoblastic cells in contact in vitro with different acrylic cements used in orthopaedic surgery. The solid and liquid components of the cements were mixed in sterile environments according to the instructions provided by the manufacturers, and placed in contact with the cells 1/2, 1, 4, 6 and 24 hours after triggering of polymerization. The experimental model, that tends to define the kinetics of the release of toxic substances from acrylic cements, allowed to verify 24 hours after polymerization that the three cements had lost their toxic effects, which instead had been present to varying degrees for the first 1-6 hours. Cytomorphological observations did not reveal cellular damage, which instead was very clear when functional tests were used. PMID- 8706549 TI - Additional methods of diagnosis in the histopathology of the musculoskeletal system. AB - In recent years because of their diagnostic importance techniques of immunohistochemistry have been increasingly employed; The use of specific cellular antibodies has allowed for the recognition of tumors of mesenchymal, epithelial, muscular, endothelial, and neuroendocrine origin, and, within these classes, for the revelation of different cellular populations. These techniques together with histochemical and ultramicroscopic studies help define not only the genesis but also the stage of differentiation and the cell function, which are important factors in the prognosis of tumors. These have proven to be particularly useful for the differential diagnosis of highly undifferentiated forms in which cellular morphology differs from that of the tissue of origin. PMID- 8706550 TI - The state of the art of non-invasive diagnostics for bone densitometry. Methods of photonic absorption. AB - The authors examine densitometric diagnostics by photonic absorption for the in vivo determination of bone mass. Based on an analysis of the literature a comparison is made for values for accuracy, repeatability, and dosimetry offered by techniques such as single-energy photon absorptiometry (SPA), dual photon absorptiometry (DPA), dual-energy X-ray absorptiometry (DXA), or quantitative computer tomography (QCT). Also discussed are economic aspects and daily management, recent developments and future possibilities. None of the densitometric diagnostics examined is capable of entirely satisfying the different needs of measurement, and nearly all of the methods of photonic absorption reported here have specific indications. The coexistence of different systems will increase the priority of the definition of standardized protocols of calibration, so as to obtain densitometric data that may be compared. PMID- 8706551 TI - Cyst of the medial meniscus: observation of two cases. AB - The authors report two cases of cyst of the medial meniscus. In both cases imaging diagnostics, CT scan, or MRI allowed us to reveal the characteristic horizontal meniscal lesion that would be the primary cause of cystic degeneration of the meniscus. The two cases were treated by selective resection of the meniscal lesion and arthroscopic decompression of the cyst. PMID- 8706552 TI - Caso quiz. Tibia vara caused by focal fibrocartilaginous dysplasia. PMID- 8706553 TI - [Congenital pseudarthrosis of the tibia treated with permanent electromagnetic fields]. PMID- 8706554 TI - [Homage to Prof. M. Paltrinieri]. PMID- 8706556 TI - [Clinical and pathological analysis of patients with hepatic metastases from colorectal cancer]. AB - 74 patients with colorectal cancer were divided into group A (34) with liver metastasis and group B (40) without by retrospective analysis. Based on the "Japanese General Rules for Clinical and Pathological Studies on Cancer of Colon, Rectum and Anus", the indicators of clinicopathological S, N, P, V, L, and inf were significantly different between the 2 groups (P < 0.01). But tumor size showed no significant difference between the 2 groups (P > 0.05). In the group A, cases of poorly differentiated carcinoma and medullary type carcinoma were more than those in the group B (P < 0.01). PMID- 8706555 TI - [Clinicopathological features and surgical treatment of early gastric cancer]. AB - We analyzed retrospectively the clinicopathological data obtained from 126 patients with early gastric cancer (EGC) to elucidate the relationship between pathological patterns and lymphatic metastasis and the criteria of selecting appropriate surgery. Lymph node metastasis was 3.1% (2/65) in mucosal and 16.4% (10/61) in submucosal tumors respectively (P < 0.05). 26 patients with tumors smaller than 10 mm had no lymphatic metastasis, and 100 patients with tumors larger than 10mm in size had 8% (8/100) n1 positive, 4% (4/100) n2 positive and 7% (7/100) lymphatic duct invasion (P < 0.05). The 5-year survival rate in patients with lymphatic metastasis (79%, 15/19) was significantly lower than that in patients without lymphatic metastasis (97.2%, 104/107) (P < 0.01). It seemed that the most important prognostic factor is whether the presence of lymphatic metastasis or not. Lymphadenectomy is expected to improve the survival rate in EGC. D2 procedure is generally recommended as a standard operation for EGC, especially for the submucosal or larger lesion. Anyhow, D1 operation is sufficient for mucosal or smaller cancer, and D3 is suitable only for EGC patients with multifoci, superficially spreading type and group 3 lymph node metastasis. PMID- 8706557 TI - [Total pelvic exenteration for local advanced rectal carcinoma: analysis of 20 patients]. AB - 20 patients of total pelvic extenteration (TPE) were performed from 1989 to 1995 for local advanced rectal carcinoma. The incidence of operative complications was 65% with a mortality of 5%. 19 patients were followed up for 3-42 months, 7 patients died with an average life-span of 14.1 months. It seemed to us that the results of conservative therapy was usually disappointing for local advanced rectal carcinoma and and TPE is the better management of choice for relieving patient's symptoms and improving their survival rates. PMID- 8706558 TI - [A modified esophagogastrostomy: report of 528 patients]. AB - The nearby tissues were used to cover the esophagogastric anastomotic stoma in 528 patients with esophageal and cardiac cancer undergoing resection. After interrupted suturing of the whole layer of esophagus and stomach, the paracervicalis was anastomosed with the capsule of the thyroid gland to strengthen the anterior wall of the anastomotic stoma. In case the anastomosis is done alove the arch of aorta, a flap of pedicled plewa is produced to cover the upper frigonum. The lower ligamentum pulmonale was used to coner the anastomosis when it is performed subaoticolly. No anastomotic leakage and stenosis were found in 528 patients in the follow-up of 2-5 years. PMID- 8706559 TI - [Anaesthesia for esophageal reconstruction with abdominal viscera in non thoracotomy]. AB - Radical resection of carcinoma of the hyopharynx and cervical esophagus usually involves pharyngolaryngo-esophagectomy and esophaged reconstruction with stomach or colonic replacement. From 1983 to 1994, 139 patients underwent the operations in our hospital. Although no thoracotomy, the extensive involvement of the neck, thorax and abdomen in the operation made the anesthesia difficult. There were 66 patients whose blood pressure dropped more than 3 kPa in the course of trans mediastinum extraction of esophagus and gastric pull-up. There were injury of posterior wall of trachea in 2 patients and spleen rupture in 3. However, no anesthetic complications occurred during the operation. PMID- 8706561 TI - [Clinical studies on changes in sexual hormones and estrogen receptor in patients with gynecomastia]. AB - Levels of serum testosterone (T), estradiol (E2), and estrogen receptor (ER) in the tissue were detected by radioimmunoassay and immunohistochemical method (ABC technique) in 46 patients with gynecomastia. The results showed that rise of E2 and E2/T is a whole-body etiologic factor of hypertrophy of male breast, and the positive rate of ER is an important local etiologic factor of gynecomastia. E2 and ER have aosynergism effect on onset and development of gynecomastia and cause the ductal cell hyperplasia of male breast. It should be alerted whether it can cause male breast cancer. PMID- 8706560 TI - [Computed sonography observation on the effects of cimetidine on the liver blood flow in cirrhotic patients with portal hypertension]. AB - The effects of cimetidine on the hepatic and portal vein blood flow were investigated in 20 cirrhotic patients with portal hypertension by computed sonography. Cimetidine given intravenously at a dose of 0.4g significantly increased the hepatic vein blood flow (60.2% +/- 63.6%, P < 0.01), at the same time its calculated cross section area increased by 40.2% +/- 81.3% (P < 0.01). The portal vein blood flow increased by 53.2% +/- 52.3% (P < 0.01). While its cross section area was 16.8% +/- 17.3% (P < 0.01). These results showed that the histamine H2-receptor antagonist cimetidine may dilate liver vascular bed and reduce portal blood flow resistance in cirrhotic patients with portal hypertension. PMID- 8706562 TI - [A study of relation of cardiac function and deformity degree in children with pectus excavatum]. AB - 40 children with pectus excavatum were divided into subgroups according to the FI (> 0.2 or < 0.2) and underwent M-ME examination for their cardiac function. 28 of 40 patients had their cardiac function and thoracic index re-examined after the operation. The results showed the cardiac function of children with pectus excavatum was damaged as FI of the patients was less than 0.2 and their cardiac function was getting well as their thoracic index improved. PMID- 8706563 TI - [Hemodynamic changes in portal system after transjugular intrahepatic portosystemic shunts (TIPS)]. AB - In this study portal venous flow (PVF), splenic venous flow (SPVF), hepatic artery volume (HAV), and portal pressure were measured before and after TIPS in 11 patients with portal hypertension. The results were compared with those of normal controls. In the normal controls PVF averaged 947.2 +/- 133.4ml/min, SPVF 239.6 +/- 116.3ml/min, and HAV 241.6 +/- 78.8ml/min. In the TIPS group before and after TIPS, PVF was 883.2 +/- 233.4ml/min vs. 958.7 +/- 185.2ml/min; SPVF was 448.9 +/- 111.6ml/min vs. 333.1 +/- 101.5ml/min; HVP was 225.3 +/- 122.7ml/min vs. 249.3 +/- 103.8ml/min. Portal pressure dropped from 3.94kPa +/- 0.46 to 2.52 +/- 0.60kPa after TIPS. We conclude that in patients after TIPS portal pressure drops blood flow in the truck of the portal vein increases, flow of hepatic tissue decreases, and blood flow in splenic vein decreases. Blood flow in hepatic aftery was not significantly changed. PMID- 8706564 TI - [Study on skin storage at -20 degrees C]. AB - Small pieces of fresh guinea pig or cadaveric skin, 0.3mm in thickness, were soaked in cryoprotective solution for 30 min, then were put into plastic bags and kept in -20 degrees C refrigerator. Before and after 14, 30, 45, 60, 75 days storage, the skin were sent for succinae dehydrogenase assay (modified Hershey's method) and oxygen consumption determination (microelectrolyte method). The average viabilities measured by succinated dehydrogenase and oxygen consumption were nearly 50% after 60 days storage and 30% after 75 days storage in both guinea pig and cadaveric skin. 37 times of autologous skin stored at -20 degrees C for 1-26 days were grafted on the granulation wounds with good results. Skin can be preserved viably at -20 degrees C for 45 days but not longer than 60 days. This method may be helpful for hospital which has no -80 degrees C deep freezer or liquid nitrogen container. PMID- 8706565 TI - [A comparative study of effects on bone healing and remodelling between embracing fixator and bone]. AB - The authors made their own design of sawteeth-arm embracing internal fixator (SEIF) with NiTi shape memory alloy and used for fractured femoral shaft of dogs in comparison with 316L stainless steel plate-screw system. A transverse osteotomy on both sides of femoral diaphyses was performed in 15 adult dogs. One side was plated with bone plate and the other with SEIF. Five animals in each group were killed at 4, 8, 12 weeks after operation and observed under roentgen rays examination, light microscopy and transmission electron microscopy. The results showed that the fracture healing and the course of callus remodelling were similar in these two groups, but the cortical bone remodelling underneath the fixater near osteotomized area was significantly different. Eight-week post implantation, osteocytes entered into degenerative phase were observed in cortical bone underneath SEIF, and 12-week after operation, the numbers of degenerative osteocytes increased remarkably. While in the case of bone plate, 4 week post-implantation, osteocytes entered into degenerative phase, and 8-week afterwards autolysis occurred in most of the degenerative osteocyles. In other words, the changes of osteocytes phase in bone plate side appeared earlier and more vigorous than that in the SEIF side. The authors are of the opinion that the elastic modulus of NiTi shape memory alloy is lower, the stress-shielding effect in the bone underneath SEIF is less, the axial compression stress of fracture line is kept larger and the contact of SEIF with the bone is not so close that is beneficial to the recovery of blood supply and bone remodelling. PMID- 8706566 TI - [Experimental steroid osteonecrosis in rabbits and pathologic findings]. AB - Osteonecrosis of femoral head was experimentally induced in adult rabbits by employing a combined protocol of hypersensitive vasculitis and administration of high dose corticosteroids. By using India ink introartery infusion microangiography and pathologic examination, we found that the damage of terminal arterioles and interruption of blood supply of terminal arteriole was causative pathogenesis. In the early phase, the haemopoietic necrosis and enlargement of fat cell in the bone marrow occurred. The perfused arterioles decreased, especially in the subchondrol bone. In the later phase, the osteocytes showed necrotic fibrosis in bone marrow. The perfused arterioles increased gradually and repairtive changes began. PMID- 8706567 TI - [Clinical and pathologic analysis of 33 cases of brachial plexus neurinoma]. AB - Clinical and pathological findings in 33 cases of brachial plexus neurinoma are reported. In 30 cases (90.91%) the diagnosis was proved to be correct by postoperative histological sections, and in other 3 cases (9.09%) the diagnosis couldn't be made preoperatively. The 33 cases were subjected to surgical resections and 18 (54.55%) of them still felt numbness and pain within two weeks after surgery. Sixteen cases were followed up for three months and 14 (87.5%) of them felt numbness by percussion over the incision. All the 33 cases had no brachial plexus traumatic complication postoperatively. PMID- 8706568 TI - [Modified distal osteotomy of shortening the first metatarsal for treatment of hallux valgus]. AB - The results of distal osteotomy of shortening the first metatarsal with internal fixation of shape, memory metal compression staple for hallux valgus were evaluated in 26 feet of 17 women with a mean age of 41.0 years (30-65 yrs). The average time for return to light work was 18.2 days, and to full work and normal walk 40.3 days after the operation. 16 patients (25 feet) had complete relief from pain. Only in one foot the pain was transferred under the 2nd metatarsal head. The appearance of the feet was apparently improved and the width of the forefeet was decreased in a mean 0.92cm after the operation. All of the patients were satisfied with the appearance of their feet. Radiographic analysis of feet showed that all the osteotomies united, the average angle of hallux valgus was improved from 30.1 degrees to 16.0 degrees and the mean intermetatarsal angle from 11.2 degrees to 6.5 degrees. This new technique allows no external fixation of plaster splintage and the distal fragment during healing of the osteotomy was stable. The time of bone healing was shortened and patients were allowed to bear weight at early time. PMID- 8706569 TI - [Intra-aortic balloon pumping in 48 patients with heart failure]. AB - 48 patients with heart failure were supported by intra-aortic balloon pumping (IABP) from 1985 to 1993. The disease included rheumatic (39 patients), congenital (7) and coronary hearts (2). The average time of supporting with IABP was 27 hours, 23 patients (48%) survied inclading 19 patients with postoperative lower cardiac output syndrome. Hemodynamics was improved. IABP increased cardiac output syndrome, and hemodynamics was improved. IABP increased cardiac output (50%), cardiac index (60%), stroke volume (50%), stroke volume index (60%) and decreased left atrial pressure or pulmonary wedge pressure. The mortality was 25 patients (52%). The death causes were as follows: arrhythmia (4), renal failure (6), lung failure (2), DIC (1), and the other (12). The main complication of IABP was lower limb ischemia particularly in surgical cut-down method to establish IABP. The analysis indicated that the temporary pacing maker with IABP could prevent some arrhythmia such as frequent premature ventricular beats. Lower limb ischemia and renal failure should receive much attention because they are easy to be confused with shock symptom when IABP. PMID- 8706570 TI - [Surgical treatment of obstructive jaundice caused by hepatocellular carcinoma]. AB - Between Jan. 1991 and Dec. 1993, 7 patients with obstructive jaundice caused by hepatocellular carcinoma underwent transcholedochal cancerous embolectomy plus T tube drainage of the biliary tract, in 5 patients with hepatectomy. Detailed discussion was made an the operative procedure and the operative results. The authors indicated that intensive surgical operation is effective in relieving symptoms and prolonging life-span. PMID- 8706571 TI - [Microwave tissue coagulation for bladder carcinoma]. AB - 47 patients with bladder carcinoma were treated by microwave tissue coagulation (MTC) through urethra. The total effective rate was 100%. Among 43 patients followed-up, 17 had recurrence of the carcinoma 6-20 months after operation and received another MTC therapy successfully. This kind of treatment is indicated for tiny, multiple, scattered and reccurred tumor. It has advantages of safety, easy operation, fast effect, mild suffering. It is an ideal treatment for the patients who are intolerable or unwilling to accept major operation. PMID- 8706572 TI - [Vena iliaca externa thrombosis after open operation of the bladder and prostate]. AB - Since 1986, we have performed total cystectomy or partial cystectomy with ureteroneocystostomy in 64 cases and prostatectomy in 182 cases. 5 cases developed vena iliaca external thrombosis (VIET) or thrombophlebitis postoperatively. Three of them recovered after treatment, and 2 died of pulmonary embolism 7 and 16 days after operations. VIET is a very serious complication. Once develops pulmonary embolism, the patient may die soon. We suggested that patients be encouraged to increase limbs activities, be given medicines of antilipemic and antiatherogenic preoperatively. And during the surgical operation, patients' blood pressure should be kept steady, and hypopietic shock should be avoided as long as possible. The vena iliaca external should never be constricted when a deep hook is used for exposure or gauze is used to stop bleeding. After operation, the patient should be encouraged to exercise on bed or around bed as early as possible, so as to increase venous flow. The veins of lower limbs should be avoided by using infusion. Styptic should not be applied abusively. Infusion of dextranum-40 and orally taking of persantinum may also be used to prevent this kind of complication. PMID- 8706573 TI - [Trigeminal neuralgia caused by microvascular compression of perforation type]. AB - Of 512 patients of trigeminal neuralgia who had undergone microvascular decompression through a retromastoid craniotomy, 4 were caused by microvascular compression of perforation type. In 3 patients, the perineurium of trigeminal nerve was cut longitudinally, the nerve-tract was separated, the perforating blood vessel was pushed to the distal end, and the vessel was made to leave the nerve sensitive area and then decompression materials were put between the separated nerve-tract. The patients were pain-free and the facial sensation was preserved after operation, and no recurrence occurred in 1 to 3 years after operation. The authors realized that there is a sensitive area near the trigeminal nerve sensory root entry zone. When the blood vessel compresses the sensitive area, trigeminal neuralgia is induced, whereas the vessel decompresses the area, the pain is free. So it is a good evidence for the microvascular compression theory and it is more important than the non-perforation type. PMID- 8706574 TI - [Application of laparoscopic technique in splenic surgery]. AB - Five patients of splenic operations had been performed by laparoscopic technique. Patients' ages ranged from 19 to 47, with an average of 31. Four were males and one was female. The procedures included splenectomy 1 patient, deroofing of splenic cyst (1), and hemostasis of splenic injuries (3). The results indicated that all the procedures were successful, and the average length of postoperative stay was 6.5 days. There were no complications. The advantages of laparoscopic splenic surgery include minimal incision, less postoperative pain and rapid recovery. It should be stressed, however, that laparoscopic splenic procedures should be performed cautiously for selected indications only and by surgical endoscopists well trained. PMID- 8706575 TI - [Mediastinoscopy: value in diagnosis of middle mediastinal masses]. AB - Between 1991 and 1994 standard cervical mediastinoscopy (SCM) was performed in 22 patients with enlarged middle mediastinal nodes of unknown causes. The pathologic diagnoses were achieved in all patients. Malignancies were found in 45.5% of the patients (10/22) and benign lesions in 54.5% (12/22). An erroneous diagnosis and hence inappropriate treatment were avoided. In our experience correct diagnosis can be made by mediastinoscopy in cases of middle mediastinal nodes enlargement, which is usually impossible by routine examination. PMID- 8706576 TI - [Peripheral anterior synechiae overlying the haptics of posterior chamber lenses]. AB - To study the changes of the anterior chamber angle after intraocular lens implantation, 111 patients (130 eyes) with posterior chamber intraocular lens implantation at different post-operative periods were examined by gonioscopy. Peripheral anterior synechia overlying the position of the lens haptic (lens haptic PAS) was observed in up to 50.8% of eyes implanted with haptics vaulted anteriorly by 10%. The lens haptic PAS is easy to be identified, it is a broad based forward displacement of the peripheral iris adhesive to the anterior wall of the anterior chamber angle. The size of the PAS was of half to one o'clock position and occasionally it was of one and a half to two o'clock position. The differences between the rates of occurrence of lens haptic PAS were not significant at different post-operative periods (P > 0.05). 55.9% of the sulcus fixated and 42.3% of the capsular fixated eyes had lens haptic PAS (P > 0.05). There were more lens haptic PAS in eyes with horizontally oriented lens haptics (62.7%) than with lens haptics at vertical position (41.4%, P < 0.05). Regular gonioscopy for posterior chamber lens implantation was recommended. PMID- 8706577 TI - [A preliminary report of small incision cataract surgery by phacoemulsification]. AB - The results of 34 cases with cataract extracted by phacoemulsification were preliminarily reported. Postoperatively, they achieved corrected visual acuities > 1.0 in 24 eyes (70.59%) in one week and in 25 eyes (73.53%) after 3 months. The post-operative astigmatism showed a tendency of change from with-the-rule shifting to against-the-rule. The rate of the corneal endothelial cell loss was 13.28%. No severe intra- and post-operative complications were found. It is concluded that the small incision, slight post-operative reaction and fast visual rehabilitation inherent in phacoemulsification procedures are more beneficial to patients than the conventional extracapsular cataract extraction. PMID- 8706578 TI - [Sutureless incision for intraocular lens implantation]. AB - The technique of incisional construction, the degree of automatic closure and the results of postoperative induced corneal astigmatism were reported in a series of 50 eyes with 5.5 mm sutureless incision for 6.0 mm PMMA intraocular lens (IOL) implantation. The patients ages ranged from 8 to 72 years. In this study, I-stage implantation was performed on 13 eyes and II-stage implantation, on 37 eyes. In this group, 19 aphakic eyes had had the histories of penetrating injuries and 1 to 3 times of operations. 15 aphakic eyes had had histories of senile cataract extractions, 14 eyes were congenital cataracts and another 2 eyes were other kinds. The surgical maneuvers included an external scleral frown incision 5 mm in chord length, a scleral-tunnel dissection and a corneal internal valve incision into the corneal limbus for 1 mm and keeping horizontally 6.0 mm in width. All the implantations were smooth, incisions closed watertightly without leakage or shallow anterior chamber. The means value of induced corneal astigmatism was 0.16(0.71)D with a range of +1.00 to -1.50 D. The complications were less than that of sutured operations. The results suggest that sutureless incision be safe, reliable, self-closed tightly, less induced astigmatism and could be a better kind of incision for standard IOL implantation. PMID- 8706579 TI - [Diagnosis and management of tumors with orbito-cranial access]. AB - To identify the characteristics and extent of orbitocranial tumor for the selection of operative methods, 64 patients with the tumor were studied with clinical materials, primarily CT and magnetic resonance imaging (MRI) appearances. The lesions were found in the orbit, cranium and orbito-cranium boundary parts and their diagnoses in regard to localization and identification could be made. Based on the preoperative diagnosis of an orbito-cranial tumor, its location, character, secondary changes and operation(s) in the past, a surgical approach was designed. The resection of a tumor could be completed in once or in several times of operation. 64 operations were performed on 53 patients and other therapies were given to the other 11 patients. The operative methods included orbitotomy via anterior route, 21 times via lateral route and 30 times via transcranial route. The operative results suggest that the transcranial route be complicated and have more complications; preoperatively, the operative indication be strictly selected and intraoperatively, ophthalmologists closely cooperate with neurological surgeons. PMID- 8706580 TI - [Comparison of simple methods to screen predisposing eye of primary angle-closure glaucoma]. AB - 6,584 population over 40 years old in Canton universities and factories were screened for the predisposing eyes of primary angle-closure glaucoma (PACG). In the screening, three methods for the measurement of anterior chamber depth were used and compared and the critical limits of two simple ones of them [peripheral anterior chamber depth (PACD) and oblique illumination test with a flashlight] were evaluated. It was found that the incidence of PACG is as high as 10.0% in eyes with axial anterior chamber depth (AACD) < or = 2.0 mm, it is 6.7% in eyes with peripheral anterior chamber depth (PACD) < or = 1/4 corneal thickness (CT) and is 6.9% in eyes with iris light band ratio (ILBR) < or = 1/4. Also, we found that the incidence of PACG is as high as 77.8% in eyes with AACD < or = 1.6 mm and 100.0% in, AACD < or = 1.4 mm. The authors propose that PACD < or = 1/4 CT and ILBR < or = 1/4 can be the critical values of the two methods for the primary step of PACG screening. PMID- 8706581 TI - [Low dosage of homoharringtonine for prevention of cicatrization after glaucoma filtering surgery]. AB - To prevent the cicatrization of filtering blebs after glaucoma filtering surgery, subconjunctival injections of low dosage homoharringtonine were administered to 22 glaucomatous eyes (20 patients) postoperatively. The percentage of functional filtering blebs was 90.9% and the mean postoperative IOP was 2.04 +/- 0.68 kPa (1 kPa = 7.5 mmHg). In comparison with the control group, the differences were very significant (P < or = 0.01). The results suggest that the toxicity of low dosage of homoharringtonine be very low, it inhibit fibroblast proliferation and play important roles in the formation of the functional filtering blebs and elevation of the success rate of filtration operation. PMID- 8706582 TI - [Combined argon and neodymium: yttrium aluminum garnet laser peripheral iridectomy: a new approach in clinical practice]. AB - Laser peripheral iridectomy was performed on 200 eyes (151 cases) with primary acute or chronic angle-closure glaucoma and partly with mixed glaucoma. Following argon laser creating an iridectomy in 2/3-3/4 iris thickness, neodymium: yttrium aluminum garnet (Nd: YAG) laser shot was required to complete a patent iridectomy. This kind of procedure is simple and tends to be more preferable to Asian people with heavily pigmented and thick irides for which a perforation is difficult to be performed. The successful rate of performing a patent iris perforation was 100.0% and 95.0% of them were completed by once of shot. The intraoperative and postoperative complications of this combined procedure appeared to be significantly less than those of either one of them. 161 eyes were followed for more than 2 years. The objective criteria demonstrate that the long term therapeutic effects of this combined technique are very good. It is recommended to use the procedure extensively in the out-patient clinics. PMID- 8706583 TI - [The deterioration of refractive accommodative esotropia]. AB - 137 patients with refractive accommodative esotropia who were followed for at least 5 years were investigated. The results showed that in 23 of the 137 patients (16.8%) occurred the deterioration of esotropia that means esotropia can no longer be controlled by wearing a pair of complete corrective spectacles. The deterioration develops most likely in patients with delay of anti-accommodative therapy or with malfunction of binocular vision. The age of onset, refractive status and the visual acuity difference between bilateral eyes are not etiologic factors in the process of deterioration. We consider that early diagnosis of accommodative esotropia, timely prescription of optical correction and maintenance of normal binocular vision play important roles in preventing the deterioration. PMID- 8706584 TI - [Examination and analysis on keratometry and axis of myopia]. AB - In this study, we applied SYSTAT software package to deal with the data, measured the keratometric values and axes of 276 patients with myopia (523 eyes), analyzed on the data of keratometry and astigmatism, correlations between the myopic degree and keratometric value and between the former and length of axis, and pathogenesis of myopia. The results revealed that the vertical corneal refractive power was stronger than the horizontal value, the vertical corneal refractive power in high and superhigh myopia was greater than that in mild and medium myopia, the axial lengths are significantly different among mild, medium, high and superhigh myopia (P < 0.01), and the myopic degree is correlated with the keratometric value and axial length. We can conclude that keratometry and axis are two main factors for the formation of myopia. PMID- 8706585 TI - [Stability of refraction in post-operative two years of radial keratotomy]. AB - In order to study the stability of the refraction after radial keratotomy, we took 123 cases (245) eyes who all had continuous follow-up records in two postoperative years to observe their dynamic changes. The mean preoperative spherical equivalent refraction was -6.74 D, and the mean keratometry was 44.32 D. The mean spherical equivalent refraction which was reduced after the surgery at one week, one month, three months, six months, one year and two years was 6.77 D, 5.74 D, 5.14 D, 5.09 D, 5.14 D and 5.27 D, respectively. The decrease of the spherical equivalent refraction in our cases showed that the maximum range of myopic drop was at one week after surgery. But the myopia obviously rose again in the postoperative three months. Statistical analyses showed that the differences between one week and one month and between one week and three months after surgery were significant (P < 0.01). There were little changes in the course of postoperative six months to two years, being not significant statistically (P > 0.05). PMID- 8706586 TI - [Distribution of fibronectin in experimental traumatic proliferative vitreoretinopathy]. AB - A rat model of proliferative vitreoretinopathy (PVR) was induced by a penetrating injury at the posterior segment of an eyeball. Immunohistochemical methods (PAP) was used for observation of the distribution of fibronectin (Fn) in the model. The chief results of this study were as follows: 8 hours after the penetrating injury, a large amount of Fn was accumulated at the wound and the vitreous cavity. On the third and 7th day after trauma, fibroblasts proliferated at the wound where Fn was located and grew into the vitreous cavity. In 14-28 days after the injury, being closely related to the proliferative fibrous tissue, Fn was distributed among the fibroblasts and the newly synthesized collagen fibers. Macrophages are the main inflammatory cell of traumatic PVR and there was rich in Fn at the surfaces of the macrophages. The results indicate that Fn is the major component in the matrix of traumatic PVR and it possibly plays an important role in the development of PVR. PMID- 8706587 TI - [An experimental study on HSV-1 corneal latency by in situ nucleic acid hybridization]. AB - An experimental study was performed on animal models for evaluation of the possibility of herpes simplex virus-1 (HSV-1) corneal latency by in situ nucleic acid hybridization. 20 normal New Zealand white (NZW) rabbits were used, and 3 x 10(6) PFU/ml of McKrae strain HSV-1 was inoculated bilaterally into the corneal stroma in 14 rabbit eyes. Of the 28 eyes, 22 developed typical herpes simplex keratitis (HSK). On the postoperative 60th day, 4 corneas with latent infection were transplanted into one unilateral eyes of each 4 non-infected NZW rabbits respectively and removed 2 weeks postoperatively. The corneal buttons were individually detected for the presence of HSV-1 antigen and nucleic acid sequences by using clonal IgG HSV-1 antibody and biotinylated HSV-1 DNA probe respectively. The results showed that the HSV-1 DNA sequences retained only within the corneal stromal layer with negative HSV-1 antigen staining. These results strongly suggest that the cornea be capable of harboring latent HSV-1. PMID- 8706588 TI - [An experimental study of corneal allograft rejection with cytotoxin IL-2-PE40]. AB - We used a new immunosuppressive agents interleukin (IL)-2-Pseudomonas exotoxin 40 (PE40) to treat corneal allograft rejection, using the Wistar rat model of heterograft-heterotopic graft rejection. 36 rats were divided randomly into the treatment and the control groups. On 2, 5, 7, 10 and 14 days after the transplantation, the transplanted corneas were removed for pathological examinations. The peripheral blood was collected for the analysis of T cell subgroups and T lymphocyte colony forming unit (T-CFU) assay. These results indicate that IL-2-PE40 can delay the development of corneal graft rejection and significantly reduce the percentage of T-helper cells. On the other hand, IL-2 PE40 can weaken the gathering capacity of the peripheral T-cells. PMID- 8706589 TI - Resistance and cross-resistance in rabbits to adults of three species of African ticks (Acari: Ixodidae). AB - Resistance to Rhipicephalus appendiculatus, Amblyomma variegatum and Amblyomma hebraeum was investigated in the laboratory by infesting rabbits with adults of each of the three species followed by homospecific or heterospecific secondary infestations. Significantly lower female engorged weights and egg mass weights were taken as evidence of protective immunity. Following a single infestation with adults, rabbits developed homospecific protective immunity (resistance) to only R. appendiculatus and A. hebraeum; primary infestation with A. variegatum did not protect against secondary infestation with the same species. There was no cross-resistance (heterospecific protective immunity) between the species except for one-way protection between R. appendiculatus and A. variegatum; primary infestation with R. appendiculatus protected against secondary infestation with A. variegatum, but not vice versa. The results from ELISA did not indicate any correlation between serum antibodies to soluble antigens from salivary gland extracts and protective immunity. Post-infection sera from rabbits infested with each of the three species reacted strongly to their respective salivary gland extracts. Despite the high reactivity of A. variegatum serum with salivary gland antigens from all three species, A. variegatum-infested rabbits did not show any homospecific or heterospecific-immunity; on the other hand, although R. appendiculatus serum did not react positively to A. variegatum antigens, infestation with R. appendiculatus protected against a subsequent A. variegatum infestation. PMID- 8706590 TI - Management of chronic pain. Part I. AB - Chronic pain is associated with substantial psychosocial and economic stress, coupled with functional loss and various levels of vocational dysfunction. The role of a pain center is to focus on chronic pain in a multidisciplinary, comprehensive manner, providing the patient with the most effective opportunity to manage his or her chronic disease syndrome. This article focuses on methods to manage many types of chronic pain and describes a broad range of pharmacologic and nonpharmacologic interventions and options available to the patient. Part I of this two-part monograph describes pharmacotherapeutic interventions and regional nerve blocks. Part II focuses on psychologic assessment and treatment and physical therapy. A multimodal management strategy offers patients the greatest improvement potential for specific chronic pain syndromes. Cognitive and behavioral therapies and physical therapies are described. This combination of therapies may provide patients with the skills and knowledge needed to increase their sense of control over pain. The integration of appropriate pharmacotherapeutic regimens, neural blockades, physical therapy, and psychologic techniques maximizes a patient's effectiveness in dealing with chronic pain. Three case studies are presented in Part II. PMID- 8706591 TI - The 4S study. Implications for prescribing. AB - This article discusses various aspects of cholesterol-lowering therapy using the HMG-CoA reductase inhibitor simvastatin in the light of the large Scandinavian Simvastatin Survival Study (4S). In 4S, patients with proven coronary heart disease (CHD) and plasma total cholesterol > 5.5 mmol/L (212 mg/dl) despite dietary measures received statin therapy or placebo for > or = 5 years. A significant mortality reduction was accomplished in those receiving the statin. Moreover, a significant decrease of nonfatal myocardial infarction and requirement for coronary bypass surgery or angioplasty was demonstrated, which will contribute to the cost-effectiveness of this well tolerated therapy. Plaque stabilisation and improvement of endothelial function are thought to be mediators of this therapeutic success. Responsible drug prescription in the post-4S era may result in the recognition and treatment of more patients with CHD. This is likely to be more beneficial than exhaustive efforts to completely achieve the goals of the most strict guidelines in the individual patient. In patients who carry the highest absolute risk for a recurrent event, aggressive drug therapy may be most justified. Reluctance to initiate lipid lowering drug therapy in patients with proven CHD should now be disputed. PMID- 8706593 TI - Comparative clinical pharmacology and therapeutic use of bisphosphonates in metabolic bone diseases. AB - The bisphosphonates are a class of synthetic compounds used in the treatment of various metabolic bone diseases, including hypercalcaemia of malignancy, Paget's disease, postmenopausal osteoporosis and corticosteroid-induced bone loss. Although there have been numerous studies comparing first, second and third generation bisphosphonates with placebo, there has been a paucity of comparative research investigating the effectiveness of these substances with other pharmacological agents. Still, the available evidence indicates that the bisphosphonates are well tolerated and effective therapeutic agents for various metabolic bone diseases. It seems certain that within the next 5 years, this class of drugs will emerge as one of the foremost options for treating Paget's disease and osteoporosis. PMID- 8706592 TI - Macrolide antibiotics in paediatric infectious diseases. AB - Erythromycin and other macrolides have enjoyed a renaissance in the 1970s, 1980s and 1990s secondary to the discovery of "new' pathogens such as Chlamydia, Legionella, Campylobacter and Mycoplasma spp. Erythromycin is an important therapeutic agent in the paediatric age group for several reasons: (a) it exhibits proven efficacy for a wide range of infections (upper and lower respiratory tract infections, skin/skin structure infections, prophylaxis of endocarditis/acute rheumatic fever/ophthalmia neonatorum and pre-colonic surgery, campylobacteriosis, chlamydial and ureaplasmal infections, diphtheria, whooping cough, streptococcal pharyngitis) and gastrointestinal (GI) dysmotility states; (b) intravenous formulations are widely available; and (c) it is available in a number of formulations as a generic product, which is likely to result in significant cost savings. Nevertheless, erythromycin and similar earlier macrolides are characterised by a number of drawbacks including a narrow spectrum of antimicrobial activity, unfavourable pharmacokinetic properties and poor GI tolerability. Newer macrolides such as clarithromycin and azithromycin are useful in serving the needs of paediatric patients who are erythromycin-intolerant or who have infections caused by organisms that are intrinsically erythromycin resistant, or for which a high percentage of strains are resistant (e.g. Haemophilus influenzae, Helicobacter pylori, Mycobacterium avium complex). In addition, these newer macrolides may be considered as alternatives to oral amoxicillin-clavulanic acid, second or third generation cephalosporins, or erythromycin plus sulphonamide in this patient population. Selection between specific macrolides and between macrolides and other antibiotics in the paediatric population is likely to depend, at least for the immediate future, on separate comparisons of product availability, cost, effectiveness and tolerability profiles. PMID- 8706594 TI - Causes and problems of nonresponse or poor response to drugs. AB - The lack of (or poor) response to drugs has very important medical, social and economic consequences today. An unsatisfactory response to treatment implies not only a worsening or prolongation of the pathological state, but also prolonged hospitalisation, longer withdrawal from social and active life, and waste of community resources. Poor response to drugs has many and varied causes, among which 4 major categories can be identified: pharmacokinetic, cellular, genetic and medical. After a brief description of the terminology, the phenomena of tachyphylaxis, tolerance and drug resistance are analysed and critically described on the basis of the available evidence. Whenever possible, alternative or operative behaviours aimed at reducing the incidence of the above phenomena are commented on. On the question of medical causes, irrational prescriptions and poor compliance with the prescribed regimen stand as two major issues whose prevalence is worrying. In order to limit the waste of professional responsibility and economic resources, efforts aimed at reducing the present insufficiencies are needed at organisational and educational levels. PMID- 8706595 TI - Current drug treatment guidelines for epithelial ovarian cancer. AB - The management of ovarian cancer is developing rapidly with the improvements in specialist multidisciplinary care. Most patients present with advanced disease and require careful surgical debulking followed by platinum-based chemotherapy. The recent introduction of paclitaxel appears to have improved the prognosis of advanced ovarian cancer by an increment that is similar in size to that seen after the introduction of cisplatin in the 1970s. Further clinical trials are required to define the optimum combination and dose of the platinum and taxane analogues, and to establish the role of the many new agents currently undergoing clinical testing. Useful chemotherapy for second-line treatment in platinum refractory patients is now available, which, combined with more aggressive surgical management, is leading to modest improvements in survival. Improvements in supportive care have enabled increasingly intensive chemotherapy to be given safely. Bone marrow support and inhibitors of specific organ toxicities are likely to be incorporated into treatment protocols over the next decade. The impact of these treatments on patients' quality of life and the economic consequences of a more active approach to management will require careful evaluation. PMID- 8706597 TI - Secnidazole. A review of its antimicrobial activity, pharmacokinetic properties and therapeutic use in the management of protozoal infections and bacterial vaginosis. AB - Secnidazole is structurally related to the commonly used 5-nitroimidazoles metronidazole and tinidazole. These drugs share a common spectrum of activity against anaerobic micro-organisms and they appear particularly effective in the treatment of amoebiasis, giardiasis, trichomoniasis and bacterial vaginosis. Secnidazole is rapidly and completely absorbed after oral administration and has a longer terminal elimination half-life (approximately 17 to 29 hours) than commonly used drugs in this class. in patients with intestinal amoebiasis or giardiasis, clinical or parasistological cure rates of 80 to 100% are achieved after treatment with a single dose of secnidazole 2g (30 mg/kg in children), similar to the response rates achieved with multiple dosage regimens of metronidazole or tinidazole. Patients with hepatic amoebiasis appears to respond well to 5- to 7-day therapy with secnidazole, but the efficacy of this drug regimen requires further evaluation in larger numbers of patients. After administration of a single dose of secnidazole, parasitological eradication was achieved in approximately 92 to 100% of patients with urogenital trichomoniasis. Patients with bacteria vaginosis respond at least as well to a single dose of secnidazole as to single-dose tinidazole, or single- or 7-day treatment with metronidazole; clinical improvement and/or microbiological evidence of cure was attained in approximately 59 to 96% of patients. In the clinical trials reviewed, secnidazole was well tolerated; most adverse events were gastrointestinal in nature and did not require treatment intervention or withdrawal from therapy. In summary, available evidence suggests that secnidazole is as efficacious as other 5-nitroimidazole drugs in the treatment of protozoal infections and bacterial vaginosis. The convenience and ease of administration associated with single-dose therapy, combined with a good tolerability profile, make secnidazole a suitable option to other single-dose treatments and an attractive alternative to multiple dosage regimens with other drugs in this class. PMID- 8706596 TI - Itraconazole. A reappraisal of its pharmacological properties and therapeutic use in the management of superficial fungal infections. AB - Itraconazole is an orally administered triazole antifungal agent. Its spectrum of activity includes dermatophyte, dimorphic and dematiaceous fungi, yeasts, and some moulds. In clinical trials, mycological cure was attained in approximately 70 to 80, > or = 70 and > or = 80% of patients with, respectively, fingernail and toenail onychomycosis (200 mg/day for 3 months), dermatophytosis (100 mg/day for 2 to 4 weeks) and vaginal candidiasis (400 mg/day for 1 day or 200 mg/day for 3 days). Approximately 20 to 30% of patients with onychomycosis may relapse after completion of therapy; relapse rate data are limited for the other indications. Recently developed intermittent regimens of itraconazole (400 mg/day for 1 week per month for 3 to 4 months) appear to have similar efficacy to standard regimens in the treatment of onychomycosis. Shorter, higher dosage itraconazole treatment regimens (200 or 400 mg/day for 1 week) are also beneficial in dermatomycoses. Discrepancies and limitations of study design hamper conclusions about efficacy relative to other antifungal drugs. Newer intermittent and short course higher dosage itraconazole regimens have also not been evaluated in comparative studies. Available studies show that the efficacy of itraconazole appears to be greater than that of griseofulvin, but similar to or lower than that of terbinafine in patients with dermatophyte onychomycosis or cutaneous fungal infections. Moreover, the efficacy of itraconazole may be similar to or lower than that of fluconazole in the treatment of cutaneous mycoses. Comparative data from patients with acute vaginal candidiasis suggest that itraconazole is at least as effective as intravaginal clotrimazole and oral fluconazole, and superior to intravaginal econazole. These results require confirmation. Prescription-event monitoring data indicate that itraconazole is generally well tolerated. Gastrointestinal disturbances, dizziness and headache occur most commonly; liver toxicity has been rarely described. Its usefulness in some clinical situations may be limited because of its ability to interact with various therapeutic agents. In conclusion, itraconazole along with other established agents should be considered a first-line treatment for patients with extensive or recalcitrant cutaneous fungal infections, mixed dermatophyte and Candida onychomycosis or vaginal candidiasis. It is currently considered a second-line drug for dermatophyte onychomycosis; the use of newer intermittent itraconazole treatment regimens may, however, extend its role in the management of this condition. Although itraconazole offers greater benefit than conventional therapies (griseofulvin and ketoconazole) in terms of efficacy and tolerability, wider clinical experience is required to determine its merits relative to the newer agents, terbinafine and fluconazole. PMID- 8706600 TI - Heart disease in Africa: what is known? PMID- 8706598 TI - Lornoxicam. A review of its pharmacology and therapeutic potential in the management of painful and inflammatory conditions. AB - Lornoxicam (chlortenoxicam), a new nonsteroidal anti-inflammatory drug (NSAID) of the oxicam class with analgesic, anti-inflammatory and antipyretic properties, is available in oral and parenteral formulations. It is distinguished from established oxicams by a relatively short elimination half-life (3 to 5 hours), which may be advantageous from a tolerability standpoint. Data from preliminary clinical trials suggest that lornoxicam is as effective as the opioid analgesics morphine, pethidine (meperidine) and tramadol in relieving postoperative pain following gynaecological or orthopaedic surgery, and as effective as other NSAIDs after oral surgery. Lornoxicam was also as effective as other NSAIDs in relieving symptoms of osteoarthritis, rheumatoid arthritis, ankylosing spondylitis, acute sciatica and low back pain. Lornoxicam has a tolerability profile characteristic of an NSAID, with gastrointestinal disturbances being the most common adverse events. Limited clinical experience to date suggests that, as with a number of other NSAIDs, lornoxicam may provide a better-tolerated alternative or adjuvant to opioid analgesics for the management of moderate to severe pain. It has also demonstrated potential as an alternative to other NSAIDs for the management of arthritis and other painful and inflammatory conditions. These preliminary findings require confirmation in further comparative and long term studies. PMID- 8706601 TI - Prevalence of heart disease in school children in rural Kenya using colour-flow echocardiography. AB - In a survey using colour-flow echocardiography and a portable generator, of one thousand one hundred fifteen children, three had clinical and echocardiographic evidence of rheumatic heart disease giving a prevalence rate of 27/1000. Sixty nine (6.2%) of the children examined had trivial mitral regurgitation (TMR). Of these, three had associated trivial aortic regurgitation. Isolated trivial aortic regurgitation was not seen. Four children had isolated mild-moderate regurgitation of the pulmonary valve. Congenital heart disease (CHD) was found in two children-one with secundum atrial septal defect and one with a ventricular septal defect and pulmonary stenosis, giving a prevalence of 1.8/1000. One child had a bicuspid aortic valve and two persistent left superior vena cava. It is feasible to carry out an echocardiographic survey using a portable generator in schools where electricity is not available. The prevalence of rheumatic heart disease (RHD) is higher than previously found in Kenya. The prevalence of TMR is surprisingly high. This was associated with familial clustering reminiscent of RHD. The importance of these findings must await the results of further investigation. PMID- 8706602 TI - Predictors of intention to be sexually active among Tanzanian school children. AB - This paper presents the results from a study of human immunodeficiency virus/AIDS risk behaviour conducted among primary school children in the Kilimanjaro and Arusha regions of northern Tanzania. The study was guided by the theory of reasoned action, and the decision to have or abstain from sexual intercourse was studied. All sixth and seventh grade pupils at eighteen schools were invited to participate. A total of 2,026 pupils (mean age 14.0 years) participated (participation rate of 85%), representing a wide variety of ethnic, socio economic and urban-rural groups. Sixty-three percent of the boys and 24% of the girls reported having had their sexual debut. Attitudes, subjective norms and self-efficacy were all predictors of intention to have sexual intercourse within the next three months, but prior behaviour emerged as the strongest predictor of intention. Primary schools appear to be an important arena for HIV/AIDS education in Tanzania and it is recommended that such education be directed toward young adolescents. PMID- 8706603 TI - Ultrasound determination of foetal gender: accuracy and social implications. AB - Seventy six pregnant women were examined by ultrasound (US) for prenatal determination of foetal gender. Results were compared with those of outcome at delivery. The range of gestational age (GA) at scan was 22-40 weeks (mean 35 +/- 4.0 weeks). Forty seven of the babies at birth were boys, while twenty nine were girls. Incorrect prediction foetal gender occurred in four out of seventy-six (5.3%) comprising two males and two females giving an overall accuracy of seventy two out of seventy six (94.7%). Accuracy of gender assignment was forty five out of forty seven (95.7%) for boys and twenty seven out of twenty nine (93.1%) for girls. It is recommended that discretion should be exercised in disclosing the gender of an unborn baby, especially if it does not correspond to the preferred sex of the mother. Determination of foetal gender should not be performed on demand or as a matter of routine in developing countries because of limited resources and shortage of skilled manpower. PMID- 8706604 TI - Clinical aspects and imaging patterns with ultrasonography and endoscopy in hepatosplenic schistosomiasis in Uganda. AB - In patients with hepato-splenic schistosomiasis, a close relationship between the grade of periportal fibrosis and portal vein diameter, spleen size and presence of oesophageal varices and their grade was found. PMID- 8706605 TI - Natural history of liver cirrhosis in 116 Nigerians. AB - One hundred and sixteen cirrhotic patients were prospectively studied over a ten year period. Hepatitis B surface antigen was positive in 70% of tested patients. The cirrhotic liver was mainly macronodular and primary hepatocellullar carcinoma was associated with 63% of the patients. Half of the patients were critically ill with high incidences of ascites, jaundice and encephalopathy. Cirrhotic patients had significantly lower body temperature onycholysis and hyperpigmented palmo plantar macular areas. The mean survival time was three years from onset of the initial symptoms to death. Patients with concomitant liver cancer were usually dead within six months after onset of the illness. Gender did not substantially affect the course of the disease. The major causes of death were tumour development (63%), gastrointestinal bleeding (40%), haemoperitoneum (28%) and hepatic failure (25%). PMID- 8706606 TI - Serum zinc, selenium and copper concentration in children with allergic colitis. AB - Children with allergic colitis have low levels of zinc and selenium which is partly a reflection of their low albumin levels and/or increased utilization of zinc and selenium as antioxidants as a result of the inflammatory process. PMID- 8706607 TI - Comparison of diagnostic methods for detection of Helicobacter pylori. AB - Of two hundred Ethiopian patients with dyspepsia, multiple biopsies were taken from the antrum of the stomach. Helicobacter pylori was cultured from 85% of duodenal ulcer and in 75% of chronic antral gastritis patients. The overall Helicobacter pylori positivity was 70%. The sensitivity, specificity, positive and negative predictive values of the tests as compared to culture were as follows, respectively: direct urease test 100%/87%/95%/100%, direct gram stain 60%/98%/99%/51%, histological gram stain 66%/97%/98%/56%, Giemsa stain 100%/97%/99%/100% and Gimenez stain 100%/87%/95%/100%. It is concluded that gram staining of direct tissue smear or histology is an insensitive method in the diagnosis of Helicobacter pylori. All the other tests, are shown to be valid. Urease test is an excellent test for provision of presumptive diagnosis of Helicobacter pylori while awaiting confirmation either by culture of histology. PMID- 8706608 TI - Bronchoscopic extraction of aspirated foreign bodies in children in Harare. AB - Fifty eight children underwent rigid bronchoscopy under general anaesthesia for suspected aspirated foreign bodies. There were 36 boys and 22 girls with a median age of thirty three months. Physical examination showed decreased breath sounds and wheezing over the affected site in forty seven cases. Chest x-rays were diagnostic in eleven cases showing definite radiopaque shadow and suggestive of aspirated foreign body in thirty five (60%). Three patients had negative results of bronchoscopy (5.2%). Extraction of foreign bodies was performed by forceps under direct vision at bronchoscopy in fifty five patients. Two children had an intraoperative hypoxic arrest. Post endoscopic complications included mild to moderate croup in fifteen patients (26%), fever in four and pneumothorax in one. Rigid bronchoscopy of aspirated foreign bodies in children is a safe, effective and often life saving procedure. Morbidity is low and although mortality was 1.8% in this series zero mortality should be achievable. PMID- 8706609 TI - Aetiological considerations in the febrile unconscious child in the rainforest and arid regions of Nigeria. AB - An analysis of hospital admissions in two areas of Nigeria indicates that the burden of coma/convulsions with fever and malaria is higher in the rainforest region whereas that of bacterial meningitis (BM) and focal extracranial infection (FEI), mainly acute respiratory infections, is higher in the arid region. The burden of malaria has increased and chloroquine-resistant malaria has become a problem in clinical practice. There is the need to revise the current policy of initiation of treatment with chloroquine in severe malaria being practised in some centres; quinine would be a suitable alternative based on current trends. Co existing infections, especially the association of other infections with BM, are an important feature in both wet and arid regions of Nigeria and point to the need for "routine" diagnostic spinal taps in order to minimise the chances of a missed diagnosis of BM. PMID- 8706599 TI - Sevoflurane. A review of its pharmacodynamic and pharmacokinetic properties and its clinical use in general anaesthesia. AB - Sevoflurane is an ether inhalation general anaesthetic agent with lower solubility in blood than isoflurane or halothane but not desflurane. The low solubility and the absence of pungency facilitate rapid mask induction; the low blood solubility also expedites "wash-out' and therefore recovery from anaesthesia. Sevoflurane produces dose-dependent CNS, cardiovascular and respiratory depressant effects that generally parallel those of isoflurane. Sevoflurane is degraded by carbon dioxide absorbents to nephrontoxic (in rats) haloalkenes, although renal toxicity has not been observed in humans. Compared with other inhalation anaesthetics, negligible quantities of carbon monoxide are generated from degradation of sevoflurane by carbon dioxide absorbents. Sevoflurane has negligible airway irritant effects, which facilitates a "smooth' induction, even in comparison with halothane in paediatric patients, and makes sevoflurane especially amenable to rapid induction of anaesthesia in adults and children. Emergence, orientation an postoperative cognitive and psychomotor function recovery of paediatric outpatients is singnificantly more rapid from sevoflurane than from halothane anaesthesia. In adult inpatients and outpatients, emergence and orientation are significantly faster after sevoflurane than after isoflurane but not desflurane anaesthesia. Other recovery parameters (e.g. times to sitting, ambulation) occur at similar times after either sevoflurane or desflurane anaesthesia. Recovery of psychomotor function occurs at generally similar times after sevoflurane, isoflurane or desflurane. Compared with propofol, sevoflurane facilitates more predictable extubation times and significantly better postoperative modified Aldrete scores in outpatients, although cognitive and psychomotor recovery occurs at similar times for both agents. As a supplement to opioid anaesthesia during coronary bypass graft surgery or in those at risk for myocardial ischaemia, sevoflurane is comparable to isoflurane. Limited data suggest that it is also as useful as isoflurane for the maintenance of anaesthesia during neurosurgical or obstetric procedures. Sevoflurane is well tolerated by adult and paediatric patients during induction of anaesthesia, with a low incidence of mild airway complications (breath holding, coughing, excitement and laryngospasm). During rapid induction, it is particularly better tolerated than isoflurane or halothane. Sevoflurane has a lower potential for hepatic injury than halothane. Unlike methoxyflurane, sevoflurane undergoes minimal intrarenal defluorination, which may account for the lack of fluoride ion-induced nephrotoxicity in humans, despite elevated plasma fluoride levels after its use. In summary, sevoflurane provides for a rapid and smooth induction of, and recovery from, anaesthesia. These features combined with its favourable cardiovascular profile should make sevoflurane the agent of choice for inhalation induction in adult and paediatric anaesthesia. Although further clinical evaluation will define the role of this agent relative to that of propofol and desflurane, sevoflurane should also prove to be a valuable alternative anaesthetic agent for adults in both outpatient and inpatient surgery. PMID- 8706610 TI - Acute appendicitis in Ethiopia. AB - Over a five year period, appendectomy comprised 17.32% of emergency abdominal operations at Gondar College of Medical Hospital, Ethiopia. The mortality rate was 4.5% and the annual incidence was uniform (nineteen to twenty two cases per year) during the study period. PMID- 8706611 TI - Pattern of acute head injuries in Kuwait. AB - Of two hundred and six consecutive head injuries, 28% had fallen from heights, and 58% had been involved in road traffic accidents, mainly pedestrians (50%). Head injury preventive measures in Kuwait should be directed to road traffic accidents and falls from heights. PMID- 8706612 TI - Levels of thyroxine, TSH, thyroid volume and mental performance among Orang Asli in selected settlements in Malaysia. AB - A significant difference in the levels of thyroxine (T4), thyroid stimulating hormone (TSH) and thyroid volume among settlements at various selected Orang Asli locations is reported. The levels improved according to the level of socio economic development. No significant difference was found in mental performance by location. PMID- 8706613 TI - Prevalence of goitre in Sekota district, Ethiopia. AB - In a community based cross sectional survey in Sekota District, the overall prevalence of gross goitre was found to be 22.8% and visible goitre prevalence of 5.8%. Altitude and age correlated with goitre prevalence. Females were more affected than males. The peak prevalence was observed in highlands; 90.1% in females beyond the age of 18 years and 49% in males between the age of thirteen to eighteen years. PMID- 8706614 TI - Update on the concepts and probable role of peptidergic nerves in dentine sensitivity and pain mechanisms. AB - Both, myelinated (A-) and unmyelinated (C-) and efferent sympathetic nerves are found in the dental pulp. The myelinated nerves are sensory and respond to stimulation of dentine with external stimuli. The A-fibres may be responsible for the sharp piercing pain sensations induced by stimulation of dentine in human subjects. C-fibres respond only when the stimuli reach the pulp proper. Activation of C-fibres may be responsible for the dull, delayed pain which radiates to the rest of the face. Immunohistochemical studies have demonstrated the existence of intradental peptidergic nerves. These are the substance P (SP), calcitonin gene-related peptide (CGRP) and vasoactive intestinal polypeptide (VIP) nerves. It is suggested that peptidergic intradental nerves may play a role in increased dentine sensitivity during inflammation and dentine hypersensitivity. PMID- 8706615 TI - Trend of birthweights of African newborns: an overview. AB - Birthweight remains a good index for defining health care delivery as well as the status of the mother prenatally and during pregnancy. Birthweight is also an important assessor of neonatal viability. It varies from each community, region or nation with a 17% incidence of low birthweight in west Africa; 13.5% in east Africa; 6.8% in western Europe and 6.1% in east Asia. PMID- 8706616 TI - Ossification of the penis: report of two cases. AB - Two patients with histologically proven penile ossification are reported from Africa, for the first time, in this communication. The aetiological causes in these patients are unclear and the pathogenesis of the condition is discussed. PMID- 8706617 TI - Simulation of insanity: case report. AB - This is a case report of an accused who appeared before a High Court in Accra, Ghana and who has had no previous history of psychiatric illness and no family history. He was sent to the Accra Psychiatric Hospital "for observation and report." A diagnosis of feigned madness or stimulation of insanity was made purely on clinical grounds. PMID- 8706618 TI - Emerging and re-emerging global microbial threats. PMID- 8706619 TI - [Importance of determining clozapine plasma level in follow-up of schizophrenic patients]. AB - Measures of plasma clozapine were carried out for 15 subjects, for up to 35 months. A positive quadratic relation (r = 0.528, n = 111, p < 0.01) has been established by using different samples at different times from each patient. From 500 mg/day this relation point to forecast an important growth of clozapine plasma concentrations. The analysis of individual data shows a large variability for the individuals and a relative stability in plasma levels for a subject having one posology. The repetition of the dosages attests the compliance. A case of epilepsy, related to an important clozapine level, is presented. So, in practice, even if neither non linear pharmacokinetics, nor genetic polymorphism have been demonstrated, over a posology of 500 mg/day, it seems interesting to proceed a clozapine dosage before increasing the posology and so reduce the comitial seizure risk. PMID- 8706620 TI - [Interobserver variation of the Global Assessment Scale. Study of 59 patients at admission to a general adult psychiatric unit]. AB - In France, an analysis of the evaluation of psychiatric activity was done by a work group representing thirteen hospitals spread over the entire country. The group just recently published their final report in the form of an experimentation protocol for the description of psychiatric activity. The proposed methodology leads to some modifications of the existing data collection based on individual patient information sheets "fiche par patient", more particularly concerning new items such as the Global Assessment Scale (GAS or Axis V of DSM III-R): our first impression was that this new type of evaluation seemed to us somewhat subjective. We therefore decided to analyze the reliability of the score established by psychiatrists not familiar with its use. In the beginning the GAS score was evaluated in clinical cases (patients seen at their admission by two or three psychiatrists and scored afterwards independently), then in theoretical cases (34 cases types taken from the Health-Sickness Rating Scale of Luborsky) with the objective to improve the consensus between clinicians. RESULTS: The clinical case studies made in an adult psychiatry pavilion permitted evaluating 59 patients at admission, of which 25 by three judges and respectively 40, 28 and 41 by pairs. The global accuracy of the score obtained with the clinical cases seems satisfactory for a discipline such as psychiatry (Kappa of Cohen's coefficient = 0.51, p < 0.001 for 25 patients by three judges--Kappa = 0.45, p < 0.001 for the 109 pairs of evaluations). The more detailed study of the classes obtained a satisfactory consensus for the scores below 40 and above 60 (K = 0.46 to 0.59) but not entirely satisfactory for the scores in the mid range of the scale between 40 and 60 (K = 0.22 to 0.29) which represents 39% of the patients in the study; this is improved by grouping into larger classes this part of the scale and it is therefore advisable to use it that way. For the 34 theoretical cases taken from the Luborsky HSRS evaluated independently by three judges the results obtained are clearly not as good (global Kappa = 0.29, p < 0.01). That is perhaps due to the fact that these "case types" don't use the descriptive patient models generally used in France. Keeping in mind that the mid range of the scale should be interpreted prudently, the GAS scale can nevertheless be credited with a certain global objectivity in the case of neutral study. Would it be the same if the GAS study could influence the allocation of resources? Taking into account all the clinical practices in mental health, the question of how all the data of this type evaluation could be recorded and updated should also examined. PMID- 8706621 TI - [Comorbidity in a sample of hospitalized depressed patients]. AB - In a retrospective study of 133 in-patients with a major depressive disorder, we first described the associated psychiatric (axis I and axis II) or somatic diseases (axis III). Then, we described the patients and therapeutical practices related to comorbidity. The results of this study showed that comorbidity concerned 89% of the patients. The mean number of diagnosis was 2.5 per patient (sd = 0.88). 33% of subjects had an associated diagnosis on axis I; 57% of subjects had an associated diagnosis on axis II; 47% of subjects had an associated diagnosis on axis III. Number of drugs prescribed was positively correlated to age (p < 0.05) and hospitalization duration (p < 0.01). Comorbidity was significantly more frequent in men than in women (p < 0.05). There was a positive correlation between comorbidity and the number of psychotropic drugs prescribed (p < 0.05). Compared to other subjects, subjects with an associated personality disorder were younger (p < 0.01), received more psychotropic drugs (p < 0.05) and less somatotropic drugs (p < 0.02). Compared to other subjects, subjects with an associated somatic disorder were older (p < 0.01), received more drugs (p < 0.01), in particular somatic drugs (p < 0.001). Compared to other subjects, subjects with an associated suicide attempt were younger (p < 0.001). However, these results should be considered with care because the numerous methodological limitations of this study, in particular the lack of standardized interviews and the lack of power. PMID- 8706622 TI - [Depression and calcinosis of the basal ganglia: apropos of a case]. AB - Idiopathic basal ganglia calcifications is a heterogeneous entity characterized by the association of bilateral and symmetric calcifications of the basal ganglia and the dentae nuclei in the cerebellum, polymorphous neuropsychiatric symptomatology. It is rare, of unknown etiology and often hereditary. We report the case of a 48 years old man, who presented with melancholic depression which has been steadily worsening for more than a year and which had led to a serious suicide attempt. Clinical observation revealed apragmatism, affective dullness and impoverished spontaneous expression; the theme of incapacity predominated but guilt feelings were absent. Neurologic examination was normal except for extra pyramidal signs. CT-scan revealed the presence of bilateral and extensive calcifications of caudate nuclei, lenticular nuclei and thalamus, which indicated a diagnosis of idiopathic basal ganglia calcification characterized by depression. Moreover, MRI revealed an hypersignal of the white periventricular substance. Phospho-calcic results and the parathormone dosage were normal. Psychometric tests showed that although intellectual capacity seemed to be intact, it was not utilised in practice. Unresponsive to antidepressants and sismotherapy, the patient is still the same one and a half year later, and needs to remain in an institution. Subsequent tests indicate that intellectual deterioration has begun. This case poses the problem of the relationship between depressive syndrome and basal ganglia lesions. We look at studies bearing on this subject and on other clinically similar syndromes (athymhormia, loss of psychic self-activation), in which the interruption or cortico-striato-pallido-thalamo cortical circuits (particularly the limbic loop), is a physiopathologic mechanism currently invoked. PMID- 8706623 TI - [Pharmacologic treatment of bulimia]. AB - The rationale for pharmacological treatment of bulimia nervosa is summarized and a review of controlled therapeutic trials shows contradictory results. A number of antidepressant agents (tricyclics: imipramine, desipramine, amitriptyline; IMAO: phenelzine, isocarboxazide; trazodone; fluoxetine) appear more effective than placebo in double-blind controlled trials of 6 to 16 weeks. In similar studies, other antidepressants (mianserine, fluvoxamine) are ineffective. Improvement reported is often incomplete and the low percentage of patients totally abstinent at the end of treatment appears of poor pronostic value for long-term outcome. Methodological limitations of existing studies are discussed, and some psychopathological factors to consider in the assessment of therapeutic response are proposed. PMID- 8706624 TI - [Phototherapy in psychiatry: clinical update and review of indications]. AB - Phototherapy introduced in 1984 by Rosenthal as a treatment for SAD (Seasonal Affective Disorder) is the first therapeutic answer to season-related psychopathology. Findings in chronobiology have largely contributed to pathophysiological theories of disorders in the internal circadian system. Actual researches on the etiology of SAD covers fields as retinal deficiency (i.e. disorder of photoreceptors), phase disturbance of the internal circadian rhythms given by internal oscillators and neuroendocrinologically drived disorders, supposing that melatonin is the main mediator of human circadian systems in the CNS. Disorders of the neurotransmitters are an other explored cue. Recent longitudinal studies show a prevalence of seasonal depressive symptoms in general population up to 10%. In populations treated for depression the prevalence of SAD is up to 20%. The SAD sex-ratio (women/men) of 3/1 is found repeatedly. Above 55 years SAD get rare. Effectiveness of phototherapy is showed in nearly all controlled studies. Bright light for patients with mild SAD appears to be most effective as is also the authors clinical impression through the practice of phototherapy in Geneva since 1991. A true placebo for bright light is still to be found according to enable evaluation of potentially important impact that unspecific therapeutic factors may trigger in phototherapy. Actually possible new indications for phototherapy are being explored: bright light for non seasonal depression has been tested with features with SAD; effectiveness in bulimia has been suggested and recently sleep disorders in psychogeriatric patients have been improved. Non seasonal circadian disorders such as jet lag might be sensitive to light. PMID- 8706625 TI - [Calcium antagonists and lithium in preventive treatment of manic-depressive disorder]. AB - Recent advances in knowledge about calcium's role as an intracellular regulator allow to broaden understanding of possible pathophysiologic mechanisms in affective disorders. An hypothesis emerge that bipolar illness arise from disorders in calcium-regulated functions. Given this hypothesis, some authors propose to describe the common profiles of action of the different mood stabilizers: all the neural mechanisms that are hypothesized to explain various psychopharmacological treatments of bipolar illness involve functions that are critically controlled by calcium. Moreover, in every instance, a known action of lithium on calcium-dependent processes could account for lithium's prophylactic results. Similarities exist between the action of lithium and calcium antagonists like verapamil and nimodipine. From these considerations the hypothesis emerge that calcium antagonists could be an alternative pharmacological agent in the maintenance treatment of bipolar illness. Calcium antagonists have been found useful in this indication by a number of investigators. Given the safety and relative lack of side effects of calcium channel blocking agents, their potential efficacy in mood disorders, it is concluded that calcium antagonists may be an alternative choice in prophylactic treatment for bipolar illness, especially in patients who cannot be treated with lithium or carbamazepine. There is evidence for using verapamil at 240 to 320 mg a day, in 2 to 4 times. Some studies suggest that the association of lithium with calcium antagonist resulted more effective than lithium alone or calcium antagonist alone in the reduction of episodes of affective disorders. However, concomitant administration of a calcium channel antagonist and lithium present adverse interactions (lithium toxicity, cardiovascular accidents), probably because of synergistic toxic effects. Therefore, authors advise care in monitoring patients receiving the combination of these medications. PMID- 8706626 TI - [Thyroid function in depressed patients]. AB - This preliminary report compares the FT3, FT4, TSH basal levels and FT4/FT3 ration of depressed patients (DSM III-R criteria) with those of a healthy control group. Authors have also studied thyroid parameters in function of some clinical depression data (polarity, intensity and endogenous character) and other factors as age and sex. 81 depressed patients (31 men, 50 women), with mean age of 44.85 years were studied. 44 patients suffered of an endogenous depression and 37 of a non endogenous depression (Newcastle criteria). 60 patients had an unipolar depression while 21 patients had a bipolar depression. The control group was constituted of 36 healthy subjects (20 men, 16 women), with mean age of 40.94 years. There is no significant difference between the two groups for sex and age, besides the different size of the two groups. FT3 mean of depressed patients was 4.39 pmol/l. There was a significant difference between unipolar group FT3 mean (4.51 +/- 1.01 pmol/l) and bipolar FT3 mean (4.03 +/- 0.91 pmol/l; t = 2.02, p < 0.05). Depression intensity was correlated negatively to FT3 mean (r = -0.23; t = 2.10, p < 0.005). FT4 mean in the depressed group was 12.62 +/- 4.14 pmol/l. The only significative result for FT4 was its correlation to TSH levels (r = -0.36; t = 3.43, p < 0.001). TSH mean in depressed patients was 1.43 +/- 0.85 microIU/ml. When we have compared these results with those of control group we concluded that FT3 and TSH levels were significantly lower in the depressed group (FT3: t = 4.83, p < 0.0001; TSH; t = 2.44 p < 0.02) and that FT4 was slightly but not significantly increased in the depressed group. FT3 decrease and the slight FT4 increase in depression may be the consequence of a metabolic deviation of FT4 transformation into FT3. Its link with intensity and polarity of depression suggests that it can be considered as a biological marker of this disease. PMID- 8706627 TI - [Comparative study of substance dependence comorbidity in bipolar, schizophrenic and schizoaffective disorders]. AB - Epidemiological studies such as the Epidemiological Catchment Area survey have shown that bipolar or schizophrenic patients are especially prone to display a comorbid substance use disorder. These studies have demonstrated that this comorbidity condition constitutes a major mental health problem owing to its high frequency. The aim of the present study was to assess the prevalence of a comorbid substance use disorder in a sample of psychotic patients, and to compare the pattern of street drug use in schizophrenic, schizoaffective and bipolar patients. Comorbidity of illicit substance use disorders was assessed with the Composite International Diagnostic Interview in 92 consecutive patients fulfilling the DSM III-R criteria for bipolar disorder (BP, n = 40), schizophrenia (S, n = 38) and schizoaffective disorder (SA, n = 14). The lifetime prevalence for any substance use was 25% in the total sample, and did not differ significantly between the three groups, although a higher prevalence was found in SA (BP: 20%, S: 23.7%, SA: 42.9%, NS). The current prevalence (previous six months) was 14.1%, in the total sample (BP: 17.5%, S: 7.9%, SA: 21.4%, NS). In the three diagnostic groups, the most commonly used drug was cannabis, followed by opiates and cocaine. These results do not confirm that schizophrenics might preferentially display abuse or dependence on psychostimulants, and highlight the possible role for the drug choice of the availability of the various illicit drugs in the geographical environment of the subject. Nearly half patients (47.8%) have a lifetime history of abuse or dependence on at least two different drugs. Age at onset of substance use disorder was earlier than or concomitant to that of schizophrenic and/or mood symptoms in most patients. This chronological pattern was the same in the three diagnostic groups. Clinical variables (age at onset, age at first hospitalization, number of hospitalizations) and sociodemographic variables (age, sex, educational level, marital and occupational status) did not significantly differ between patients with a lifetime history of drug abuse or dependence and those without. Patients presenting with a current abuse or dependence were younger than those without. These results confirm in a French sample of schizophrenic and/or mood disordered patients the high frequency of the comorbidity with substance use disorders. PMID- 8706628 TI - The nature and function of putative gonadotropin surge-attenuating/inhibiting factor (GnSAF/IF). PMID- 8706629 TI - Regulation of ovarian follicular development in primates: facts and hypotheses. PMID- 8706630 TI - Peptide signaling in human placenta and membranes: autocrine, paracrine, and endocrine mechanisms. PMID- 8706631 TI - Antidepressants and hypothalamic-pituitary-adrenocortical regulation. PMID- 8706632 TI - Enhancement of the macrophage oxidative burst by immunoglobulin coating of respirable fibers: fiber-specific differences between asbestos and man-made fibers. AB - The ability of long amosite asbestos fibers (LFA), vitreous fibers (MMVF 21 and CODE 100/475), and ceramic fibers (silicon carbide and RCF 1) to stimulate superoxide production in isolated rat alveolar macrophages is examined. The cells were exposed to both naked fibers (uncoated) and fibers coated with rat immunoglobulin (IgG), a normal component of lung lining fluid. The affinity for IgG of the various fibers was assessed by quantifying the binding of 125I-labeled IgG onto the fiber surface. Naked fibers stimulated a modest release of superoxide anion from alveolar macrophages, which was not obviously dose dependent. When IgG was adsorbed onto fibers of MMVF 21 and RCF 1, there was a dramatic increase in superoxide release, which correlated well with their high affinity for IgG.IgG-adsorbed code 100/475 and silicon carbide whiskers (SiCW) stimulated only modest superoxide release, and the fibers showed a correspondingly poor affinity for the opsonin. Conversely, the adsorbed fibers of LFA, generated a dramatic increase in superoxide release from the macrophages, despite a relatively poor adsorption of IgG. This study demonstrates the potential for components of the lung lining fluid to modify the response of alveolar macrophages to respirable natural and man-made fibers. It also draws attention to fiber-specific differences in adsorptive capacity and subsequent biological activity between these fiber types in vitro and, by implication, in vivo. PMID- 8706633 TI - Elevated levels of IL-6, INF-gamma, and TNF-alpha in mice in response to cotton dust are modulated by anti-TNF-alpha antiserum. AB - Acute pulmonary neutrophilic inflammation triggered by cotton dust exposure is one of the features of organic dust syndrome. Studies with a mouse model have reproduced the inflammation and have shown the presence of tumor necrosis factor alpha (TNF-alpha) in the bronchoalveolar lavage (BAL) fluid of mice following a 3 h exposure to respirable cotton dust particles. A cover glass technique for cytospin samples of BAL cells resulted in a 42-fold increase in cell count, with 76% neutrophils, 13% lymphocytes, and 10% macrophages, after cotton dust exposure. Immunohistochemical staining of lung specimens with anti-TNF-alpha antiserum revealed TNF in the cells surrounding pulmonary airways and vessels. Cotton dust exposure resulted in elevated TNF-alpha, IL-6, and INF-gamma in BAL fluid, INF-gamma and IL-6 in serum. Administration of anti-TNF-alpha antiserum prior to the organic dust exposure resulted in a marked attenuation of the pulmonary inflammatory cell response, as well as decreased IL-6 and TNF-alpha levels in BAL fluid and decreased IL-6 and INF-gamma in serum. These results indicate TNF modulation of the dust-induced toxic alveolitis and cytokine production. PMID- 8706634 TI - Down-regulation of canine airway mast cell function following exposure to ozone in vivo. AB - The influence of ozone on the function of airway cells involved in allergic responses is of particular interest due to the increasing prevalence of this environmental oxidant pollutant. In the present studies, the peripheral airways of Ascaris-sensitive dogs were expose to ozone (1 ppm, 5 min) or air (control) and the exposed segments were lavaged 30 min later. The kinetics and magnitude of release of PGD2 and histamine from canine peripheral airway mast cells (PAMC) was determined following in vitro challenge with Ascaris antigen or calcium ionophore approximately 4 h from the time of the in vivo exposures. Histamine content was significantly lower in PAMC retrieved from ozone- versus air-exposed segments (3.0 +/- 0.1 vs. 5.2 +/- 0.5 pg/cell). Absolute release of histamine at 20 min was decreased by 47 and 43% in ozone-exposed cells stimulated with antigen or ionophore, respectively. Maximal synthesis and release of PGD2 in response to antigen (345 +/- 22 pg/10(3) PAMC) or ionophore (1055 +/- 104 pg/10(3) PAMC) was inhibited by 32 and 55%, respectively, in cells from ozone-exposed segments. Inhibition of prostanoid synthesis was not observed in alveolar macrophages in the lavage samples, nor could decreased PGD2 be attributed to enhanced catabolism. These data indicate a differential down-regulatory influence of ozone on subsequent release of granular mediators and newly synthesized PGD2 from PAMC following brief in vivo exposure that lasts for several hours. PMID- 8706635 TI - Antioxidant depletion, lipid peroxidation, and impairment of calcium transport induced by air-blast overpressure in rat lungs. AB - Exposure to blast overpressure, or the sudden rise in atmospheric pressure after explosive detonation, results in damage mainly of the gas-filled organs. In addition to the physical damage, in the lung, injury may proceed via a hemorrhage dependent mechanism initiating oxidative stress and accumulation of lipid peroxidation products. Massive rupture of capillaries and red blood cells, release of hemoglobin, its oxidation to met-hemoglobin and degradation sets the stage for heme-catalyzed oxidations. The authors hypothesized that lipid hydroperoxides interact with met-hemoglobin in the lungs of exposed animals to produce ferryl-hemoglobin, an extremely potent oxidant that induces oxidative damage by depleting antioxidants and initiating peroxidation reactions. Oxidation induced disturbance of Ca2+ homeostasis facilitates further amplification of the damage. To test this hypothesis, groups of anesthetized rats (6 rats/group) were exposed to blast at 3 peak pressures: low (61.2 kPa), medium (95.2 kPa), high (136 kPa). One group served as an unexposed control. Immediately after exposure, the rats were euthanized and the lungs were analyzed for biochemical parameters. Blast overpressure caused: (1) depletion of total and water-soluble pulmonary antioxidant reserves and individual antioxidants (ascorbate, vitamin E, GSH), (2) accumulation of lipid peroxidation products (conjugated dienes, TBARS), and (3) inhibition of ATP-dependent Ca2+ transport. The magnitude of these changes in the lungs was proportional to the peak blast overpressure. Inhibition of Ca2+ transport strongly correlated with both depletion of antioxidants and enhancement of lipid peroxidation. In model experiments, met-hemoglobin/H2O2 produced damage to Ca2+ transport in the lungs from control animals similar to that observed in the lungs from blast overpressure-exposed animals. Ascorbate, which is known to reduce ferryl-hemoglobin, protected against met-hemoglobin/H2O2-induced damage of Ca2+ transport. If ferryl-hemoglobin is the major reactive oxygen species released by hemorrhage, then its specific reductants (e.g., nitric oxide) along with other antioxidants may be beneficial protectants against pulmonary barotrauma. PMID- 8706636 TI - Effect of high glucose on fetal lung maturation at different times in gestation. AB - Hyperglycemia has been implicated as a cause of delayed fetal lung maturation in the diabetic pregnancy. It has been previously demonstrated that high glucose levels inhibit biochemical and morphological maturation of fetal rat lung in vitro. To explore whether there is a critical period for maximal effects of high glucose, fetal rat lung explants from day 18-22 (term) were examined after culture for 44 h in media containing 10 or 100 mM glucose. Rate of choline incorporation into PC and DSPC was significantly decreased in explants derived from day 20-22 fetuses after culture with high glucose. Paradoxically, in explants derived from day 18-19 fetuses choline incorporation rate was increased. Significant decreases in total PC and DSPC were seen in the high glucose group on days 20-21; however, earlier in gestation no differences in PC and DSPC were noted, despite the higher rate of choline incorporation into PC and DSPC, suggesting an increased turnover rate. High glucose levels also resulted in significantly decreased incorporation of labeled glycerol and palmitate into PC. Morphologic analysis of right upper lobe explants revealed significant decreases in the number of type II pneumocytes and lamellar bodies per alveolar lining cell in high glucose treated explants derived from days 19-20, but not before or after that time in gestation. Prior to day 20 there were few luminal lamellar bodies in potential airspaces. By day 20, the high glucose group had significantly fewer luminal lamellar bodies, although by day 21 the difference was no longer significant. These results suggest that high glucose may have a greater inhibitory effect late in gestation and that there may be critical periods in lung development when differences in substrate availability and utilization have differing effects. PMID- 8706637 TI - Impaired surfactant synthesis in fetal type II lung cells from gsd/gsd rats. AB - The importance of intracellular glycogen for surfactant synthesis was investigated in fetal type II lung cells isolated from rats with a glycogen storage disorder, designated gsd/gsd. Compared to cells from a control Wistar strain, cultured gsd/gsd pneumocytes were glycogenrich and contained fewer and smaller lamellar inclusions. Freshly isolated cells from day 19-21 fetuses of control rats demonstrated the expected gestational rise and fall of cellular glycogen seen in intact fetal lungs. At day 20, when tissue glycogen peaks, cellular glycogen content was 48 and 70 nmol glucose/micrograms DNA in isolated type II cells of control and gsd/gsd lungs, respectively. In control cells, while active glycogen phosphorylase changed from 35 to 65% of total during 24 h of culture, glycogen fell 85%. In contrast, gsd/gsd cell phosphorylase was not activated, phosphorylase kinase activity was nondetectable, and glycogen per cell remained unchanged. [3H]Choline incorporation into total PC and disaturated PC (DSPC) was 50 and 62% lower, respectively, in gsd/gsd type II cells compared to controls in the absence of exogenous substrate. Cellular content of the surfactant-associated protein SP-A was similar in control and gsd/gsd cells at day 20, and increased 3- to 4-fold during a subsequent 24-h interval of tissue culture. These results suggest that PC synthesis is dramatically impaired in type II cells in which glycogen cannot be mobilized, but SP-A is synthesized at normal rates. This work characterizes the isolated gsd/gsd fetal type II cell and supports its use in future studies to determine the importance and relative utilization of specific nonglycogen substrates. PMID- 8706638 TI - Modulation of the anchorage-independent phenotype of human lung fibroblasts obtained from fibrotic tissue following culture with retinoid and corticosteroid. AB - Fibroblast heterogeneity has been documented in fibrotic tissue from lung and skin. Differences have been demonstrated in proliferative rates in fibroblasts derived from fibrotic lung tissue as compared to normal. Fibroblast lines derived from adult fibrotic lung tissue and neonatal normal lung tissue exhibit colony growth in soft agarose culture, whereas fibroblast cell lines from normal adult lung tissue do not. The characteristic of anchorage-independent growth is consistent with the aggressive nature of the disease and with developmental lung growth. In this study, fibrotic lung fibroblasts were exposed to growth and differentiating factors to determine whether the anchorage-independent phenotype can be modulated. The results indicate that treatment of fibrotic lung fibroblasts with retinoic acid, known to modify matrix gene expression and induce differentiation, inhibits the cells ability to form colonies under soft agarose growth. Treatment with all-trans-retinoic acid yielded the greatest effect inhibiting both IPF and neonatal lung fibroblast anchorage-independent growth approximately 90% at 10(-6) M. Treatment of IPF fibroblasts with all-trans retinoic acid also inhibited corticosteroid-induced colony growth. Modulation of the "fibrotic" fibroblast phenotype through retinoid therapy may prove beneficial as a potential therapeutic strategy. PMID- 8706639 TI - Effect of interleukin-2 and interleukin-4 on lymphocytes from peribronchial lymphatic tissue. AB - Lymphocyte activation in spleen and peribronchial lymphatic tissue (PBLT) following stimulation with T-cell mitogens and lymphokines was investigated in the hamster. Optimal mitogen-induced cell proliferation was achieved after culturing for 6 days in vitro. Interleukin-2 (IL-2) and interleukin-4 (IL-4) did not induce DNA synthesis in resting T-cells from either spleen or PBLT.IL-2 or IL 4 in combination with concanavalin A promoted splenic T-cell proliferation. In contrast, in PBLT, IL-2-but not IL-4-enhanced cell proliferation (p < .001). The findings indicate that PBLT represents an independent compartment of the immune system. Furthermore, in the hamster PBLT cells consist predominantly of IL-2 responsive cells, i.e., are of the Th1 type. Immunological pathogenesis of lung injury can therefore be studied by functional analysis of PBLT lymphocytes. PMID- 8706640 TI - Lymphocyte subsets in the endometrium of genitally normal mares and mares susceptible to endometritis. AB - The density and distribution of MHC Class II positive cells and subpopulations of lymphocytes were studied in the endometrium of genitally normal mares and mares susceptible to endometritis. In genitally normal mares, more MHC Class II positive cells were present in the epithelium and stratum compactum during oestrus than dioestrus. Significantly more CD4+ and CD8+ lymphocytes were present in the stratum compactum than in the stratum spongiosum. CD4+ lymphocytes were present in greater numbers than CD8+ lymphocytes in the stratum compactum but approximately equal numbers were present in the stratum spongiosum and in lymphoid aggregates. Occasional CD4+ and CD8+ cells were seen in the luminal and glandular epithelium. Infrequently, B cells were present in the endometrium and were not observed in the epithelium. Numbers of T and B cells did not appear to be affected by cycle stage. In mares with endometritis, the densities of CD4+, CD8+ and B cells were significantly increased. Large aggregates of lymphoid cells which contained approximately twice as many CD4+ cells as CD8+ cells were present in the endometrium of these mares and all 3 subclasses of lymphocyte were seen occasionally in luminal and glandular epithelium. PMID- 8706641 TI - Retinal detachment in horses. AB - Ten horses with partial or total retinal detachment were investigated using light and electron microscopy (TEM, SEM). Several lesions were observed and compared with normal retinal morphology. Three weeks after the initial retinal detachment, hypertrophy of the pigment epithelium with transformation of the apical microvilli could be observed. The lesions were accompanied by progressive degeneration and atrophy of the sensory retina, starting at the photoreceptor outer segments. Hypertrophy of Muller cells was a concomitant finding. Retinal detachment represents a sequel either to inflammatory processes or trauma. PMID- 8706642 TI - Effect of a warm-up on energy supply during high intensity exercise in horses. AB - The VO2(max) in racehorses is approximately double that of elite human athletes and the rate of increase in VO2 at the onset of high intensity exercise is much greater than in man. The kinetics of gas exchange are affected by a warm-up prior to exercise in humans, there being a greater aerobic contribution to high intensity exercise after warm-up. Our hypothesis was that a warm-up would increase aerobic energy delivery in racehorses during high intensity exercise. Thirteen fit Standardbred racehorses ran to fatigue at 115% of VO2(max) on a treadmill at 10% slope. Prior to acceleration, horses were exercised either for 5 min at 50% VO2(max) followed by 5 min walk, or walked for 2 min. Samples of expired gas were collected every 10 s during the run for determination of VO2 and VCO2 and measurement of maximal accumulated oxygen deficit (MAOD). Blood lactate concentration was measured 5 min post exercise. We found that with a warm-up, horses had faster kinetics of gas exchange and a greater proportion of their total energy requirement was supplied by aerobic sources. The aerobic contribution to total energy requirement with and without warm-up was, respectively, 79.3 +/- 1.0% and 72.4 +/- 1.7% (P < 0.01). There was also a higher MAOD (P < 0.01) in horses that had not been given a warm-up (mean +/- s.e.m. 34.7 +/- 2.6 and 47.3 +/- 2.6 mLO2eq/kg bwt with and without a warm-up respectively). However, there were no significant differences in total run time or estimated total energy expenditure between the 2 protocols. We concluded that during high intensity exercise to fatigue lasting 1 to 2 min, more than 70% of energy supply is from aerobic energy sources and that this contribution is even greater when the horses have received a warm-up. PMID- 8706643 TI - Regulatory significance of procaine residues in plasma and urine samples: preliminary communication. AB - Plasma and urinary concentrations of procaine and the duration of response to procaine after its administration as a local anaesthetic to horses were studied. Following injection of a clinical dose of procaine HCl (80 mg), the concentration of procaine in plasma was less than the lower limit of quantitation and unsuitable for threshold determination. Therefore, the urinary concentration of procaine was determined after injection of a dose of 5 mg procaine HCl, the highest no-effect dose (HNED) of this agent. Free unconjugated procaine in equine urine reached a peak concentration of 23.7 ng/mL, while total (unconjugated plus conjugated) procaine peaked at 37.9 ng/mL (mean urine pH of 8.5). Because a basic drug may concentrate substantially in acidic urine, a threshold concentration of 25 ng/mL of unconjugated procaine is a reasonable and conservative threshold for procaine at this time. Horses were administered abaxial sesamoid blocks containing 2% procaine HCl (40, 80, 160 and 320 mg) and 2% procaine HCl (40 and 320 mg) with epinephrine (1:100,000) in local anaesthetic experiments. There was a significant local anaesthetic (LA) effect for all doses of procaine HCl with the duration of effect ranging from 30 min (40 mg) to 60 min (320 mg). The addition of epinephrine significantly increased the duration of local anaesthesia to 180 min for a 40 mg dose and 420 min for a 320 mg dose. Because epinephrine may extend the duration of local anaesthesia beyond a reasonable period of confinement for horses before the starting time of a race, the increased LA effect following the addition of epinephrine to procaine has regulatory significance. PMID- 8706644 TI - Influence of shoeing on ground reaction forces and tendon strains in the forelimbs of ponies. AB - Strains in the superficial digital flexor tendon (SDFT), deep digital flexor tendon (DDFT), accessory ligament of the deep digital flexor muscle (inferior check ligament [ICL]) and the interosseus medius muscle (suspensory ligament [SL]) in the right forelimb of 5 ponies were measured using mercury-in-silastic strain gauges a few hours after implantation. Tendon strains were recorded at the walk with normal flat shoes, egg-bar shoes, a 7 degrees increased hoof angle accomplished by application of a heel-wedge and a 7 degrees decreased hoof angle using a toe-wedge, consecutively. Ground reaction forces were recorded with all 4 shoe types preoperatively and with flat shoes post operatively. The strain patterns of the SDFT, DDFT and SL showed a rapid increase at the beginning of the stance phase, followed by a plateau with a small incline or decline and a rapid decrease at the end of the stance phase. The SDFT had its maximal strain in the first half of the stance phase in all ponies. The DDFT and SL reached their maximal strain in the first half of the stance phase in 2 ponies and in the second half of the stance phase in the other 3 ponies. The ICL was strained maximally in the second half of the stance phase in all ponies. Averaged over all 5 ponies, the maximal strains in the SDFT, DDFT, ICL and SL with normal flat shoes were 2.4, 1.3, 5.4 and 3.7%, respectively. If an egg-bar was applied the mean peak strain in the DDFT was 0.13% lower and strain in the SL was 0.22% higher. With a heel-wedge, strain decreased in the DDFT and ICL (0.19% and 0.4%, respectively) and increased by 0.24% in the SL. A toe-wedge increased strain in the ICL by 0.8%. All changes mentioned were statistically significant (P < 0.1). The changes in tendon strain as a result of different types of shoeing correlated with changes in calculated torque's of the ground reaction force acting on the coffin joint. PMID- 8706645 TI - Tendon strain in the forelimbs as a function of gait and ground characteristics and in vitro limb loading in ponies. AB - Strains in the tendons of the m. flexor digitalis superficialis (superficial digital flexor, SDFT) and m. flexor digitalis profundus (deep digital flexor, DDFT) tendons, the accessory ligament of the deep digital flexor muscle (inferior check ligament, ICL) and the m. interosseus medius (suspensory ligament, SL) of 5 ponies were recorded at the walk and trot using mercury-in-silastic strain gauges (MISS), on a hard surface (brick pavement) and on sand. The horses were shod with normal, flat shoes. On pavement, strain in the SDFT, DDFT and SL increased significantly from the walk (2.19%, 1.15% and 3.36%, respectively) to the trot (4.15%, 1.70% and 5.78%, respectively), but that in the ICL did not change significantly (5.36% at the walk, 4.88% at the trot). Strains in the ICL and SL were higher on pavement than on sand (P < 0.1) and strains in the SDFT and DDFT were not significantly different. Tendon strain in the SDFT and SL, but not in the ICL and DDFT, increased (P < 0.1) in a pony at the walk on pavement with a rider. Post mortem loading of the same instrumented limbs revealed that the metacarpophalangeal joint could be further extended when the elbow joint was extended. The in vitro tendon strain was different from that in vivo, implying that results from in vitro limb loading tests have only limited value for assessing tendon functioning in vivo. PMID- 8706646 TI - Surgical treatment for colic in the foal (67 cases): 1980-1992. AB - Sixty-seven foals age < 150 days underwent a ventral celiotomy for colic. Of the 67 foals, 51 foals (82%) recovered from anaesthesia and 42 (63%) were subsequently released from the hospital. Three (6%) of the 51 foals were subjected to a repeat celiotomy. Long term follow-up was available on 36 foals. Twenty-nine (57%) of the 51 foals recovered from anaesthesia, were alive at least 2 years following surgery. Adhesions were identified in 8 (17%) of the foals which recovered from general anaesthesia but were subsequently subjected to euthanasia due to recurrent colic. Strangulating lesions were associated with a lower survival rate. Nineteen per cent of foals with strangulating intestinal lesions survived > 2 years following surgery, compared to 69% of foals with nonstrangulating lesions. The age of foals on admission had a significant effect on survival. Only 10% of foals less than 14 days of age survived, compared to 45.8% of foals between age 15 and 150 days. PMID- 8706647 TI - Clinical and pathological studies in horses with hepatic disease. AB - In horses with hepatic necrosis, lipidosis, neoplasia and cirrhosis, progression of the disease was studied by serial measurements of total serum bile acid concentrations and of plasma glutamate dehydrogenase (GD) and gamma glutamyl transferase (gamma GT) and by liver biopsy. Plasma ammonia concentrations were significantly elevated compared to clinically normal horses, but such changes were not always accompanied by a decline in plasma urea concentration. A fall in plasma glucose concentration carried a guarded prognosis. These were all invaluable aids in early diagnosis and throughout the disease course. The study suggests that other factors, such as hypokalaemia, alkalosis, short chain volatile fatty acids, false and true neurotransmitters, may be important in the pathogenesis of hepatic coma in the horse. PMID- 8706648 TI - Effects of equine influenza and tetanus vaccination on pulmonary function in normal and chronic obstructive pulmonary disease affected horses. PMID- 8706649 TI - Zinc phosphide poisoning in a horse. PMID- 8706650 TI - Sudden unexpected death in a horse due to a cerebral oligodendroglioma. PMID- 8706651 TI - Nonfatal subcapsular splenic haematoma in a horse. PMID- 8706652 TI - Sherlock Holmes and chemical poisons. PMID- 8706653 TI - Computed tomography (CT): a dip into the future. PMID- 8706654 TI - Radiographic and arthroscopic findings associated with subchondral lucency of the distal radial carpal bone in 71 horses. AB - Case records and radiographs of 71 horses with subchondral lucency, without radiographic evidence of fracture, located on the distal radial carpal bone were examined retrospectively. All horses had lameness and/or joint effusion referable to the carpus. Distal radial carpal bone subchondral lucency was found as a solitary lesion or as a lesion concurrent and symmetric to a contralateral distal radial carpal bone chip fracture. The lesion appeared radiographically as a lucency or shadow on the distal dorsal margin of the radial carpal bone and was most evident on the flexed lateromedial and dorsolateral-palmaromedial oblique projections. Sixty-four joints in 55 horses underwent arthroscopic surgery. Surgical findings included osteochondral fragmentation in 44 joints, cartilage fragmentation with subchondral bone softening in 17 joints and cartilage fraying in 3 joints. Corresponding third carpal bone lesions were observed in 18 joints and moderate to severe synovitis was present in 24 joints. Carpal bone subchondral radiolucency without a fracture fragment observed on radiographs indicated cartilage and bone damage. In some cases, small chip fractures (1-2 mm) were present within the cartilage debris examined at surgery, yet were not radiographically visible. Follow-up information was obtained on 50 of the operated horses and 14 nonoperated horses. Forty (80%) of the surgically treated horses returned to racing, with 34 (68%) of these horses (20 of 26 with subchondral lucency as a solitary lesion) racing at a level of competition equal to or better than the pre-injury level. Of the 14 horses which were not operated, 6 (42%) returned to racing of which only 2 (14%) raced at the same pre-injury level of competition. Radiographic evidence of radial or intermediate carpal bone subchondral lucency is an indication of cartilage and bone damage which is best treated with arthroscopic surgery. PMID- 8706655 TI - Computed tomographic evaluation of head diseases in the horse: 15 cases. AB - The rapid development of software and technology now allows a large amount of diagnostic information to be obtained from a computed tomographic examination. This imaging technique can also be usefully applied to the horse, given appropriate premises and a custom built table for accurate positioning. Computed tomography of the skull has considerable advantages over other techniques, as structures are viewed without superimposition. Fifteen cases are used to demonstrate how through high image quality (precise detail, reduction in artefacts) and objective measurement of density, various pathological changes can be analysed and exact diagnoses achieved. In particular, short measurement and examination times allow preoperative imaging under general anaesthesia giving significant information to assist subsequent surgery. PMID- 8706656 TI - A TGACG motif mediates growth-hormone factor-1/pituitary-transcriptional activator-1-dependent cAMP regulation of the rainbow trout growth-hormone promoter. AB - The mechanisms involved in the regulation of the rainbow trout growth hormone (tGH) gene promoter by the pituitary-specific transcription factor GHF1 (growth hormone factor 1), also called Pit1 (pituitary transcriptional activator 1), and cAMP have been investigated in mammalian and fish cells. The -340 to +24 5' flanking Fegion of the tGH gene focused to the luciferase gene was activated in rat pituitary GC cells and in HeLa cells cotransfected with an effector plasmid encoding rat GHFI. GC cell nuclear extracts produced four GHFI-specific footprints (sites Fl to F4) on the tGH promoter, each containing multiple W4NCAT (W, A or T) or closely related motifs. Mutational analysis performed in GC cells indicated that the proximal Fl site alone can direct transcription, but that the region encompassing the F2 and F3 sites is necessary for optimal activation and contains a TGACG motif (cAMP-response element, CRE) conferring cAMP responsiveness. The role of the TGACG motif in mediating cAMP regulation of the tGH promoter was confirmed in primary cultures of trout pituitary cells. Cotransfection studies in carp EPC cells using an effector plasmid encoding trout GHF1 demonstrated the GHF1 dependence of cAMP stimulation. Gel shift and southwestern experiments revealed nuclear proteins of 43 kDa and 30 kDa in GC and fish cells, respectively, that bind specifically to the tGH CRE, suggesting the involvement of CRE-binding-protein/activating-transcription-factor-l-related peptides in cAMP response. Incidentally, and in contrast with previous reports, we found the rat GH promoter, that lacks TGACG motifs, unresponsive to cAMP. Thus, the CAMP stimulation of the tGH gene is more similar to its human counterpart. that is also GHF1 dependent and mediated by TGACG motifs in the promoter. It is suggested that control of GH gene expression has evolved modularly, through various assortments of the same regulatory units, rather than molecularly, through innovative units. PMID- 8706657 TI - Formation of a novel enzymatic metabolite of leukotriene A4 in tissues of Xenopus laevis. AB - Leukotriene-A4, hydrolase catalyzes the final step in the biosynthesis of the potent proinflammatory mediator leukotriene B4. Previously, leukotriene-A4 hydrolase has been characterized from human, mouse and rat sources, i.e. only from mammalian species. In the present investigation, expression of leukotriene A4, hydrolase was studied in organs of Xenopus laevis. Enzyme activity was found in all nine organs tested with the highest levels in the intestine and the reproductive organs, i.e. oocytes and testes, previously unrecognized rich sources of the enzyme. No immunoreactive leukotriene-A4 hydrolase was detected in Western blots of 10000Xg supernatants of X. laevis organ homogenates, using a polyclonal antiserum raised against human leukotriene-A4 hydrolase. Likewise, Northern blot analysis of liver total RNA did not detect Xenopus leukotriene-A4 hydrolase mRNA using a human CDNA probe. These results indicate significant structural differences between the human and toad enzymes. Incubations of 10000Xg supernatants of organ homogenates with leukotriene A4 revealed the formation of a novel metabolite, denoted compound X. Conversion of leukotriene A4 into compound X was due to an enzymatic activity as judged by its protein dependence, heat sensitivity, and resistance to ultrafiltration, and this activity appeared to be linked, directly or indirectly,, to leukotriene A4 hydrolase. From data obtained by ultraviolet spectrophotometry, gas chromatography coupled to mass spectrometry, ultraviolet-induced isomerization, and comparison with a synthetic standard, compound X was assigned the structure 5S,12R-dihydroxy-6,10-trans-8,14 cis-eicosatetraenoic acid. Finally, compound X was found to exhibit contractile activity in guinea-pig lung parenchyma, apparently elicited via a leukotriene B receptor. PMID- 8706658 TI - Molecular cloning, expression and characterization of mouse leukotriene C4 synthase. AB - Leukotriene C4 synthase (EC 2.5.1.37) catalyzes the conjugation of reduced glutathione (GSH) with leukotriene A4 to form the intracellular parent of the proinflammatory cysteinyl leukotrienes. Human leukotriene C4 synthase shares substantial amino acid identity in its consensus N-terminal two-thirds with 5 lipoxygenase-activating protein and has a region (residues 37-58) that exhibits 46% amino acid identity with a domain of this protein (residues 41 -62) to which an inhibitor binds. We have now cloned mouse leukotriene C4 synthase CDNA using the polymerase chain reaction to screen a mouse pcDNA3 expression library with oligonucleotide primers based on the translated human leukotriene C4 synthase cDNA sequence. Mouse leukotriene C4 synthase cDNA is 667 bp in length, including the poly(A)-rich tail, and shows 87% similarity with the human cDNA within the open reading frame. The deduced 150-amino-acid sequence of mouse leukotriene C4 synthase (differs from the human enzyme by only 18 amino acids, of which 9 reside at the C terminus. The potential N-glycosylation site, two protein kinase C phosphorylation sites, the two cysteine residues, and the putative inhibitor binding domain (substitutions Thr4l-->Ser and Tyr50-->Phe) were conserved in mouse leukotriene C4 synthase. Northern blot analysis indicated that the leukotriene C4 synthase RNA transcript is widely distributed. The Km values for leukotriene A4 methyl ester, leukotriene A4 free acid and GSH were 7.6 microM, 3.6 microM and 1.6 mM, respectively, for purified human recombinant enzyme, and 10.3 microM, 2.5 microM and 1.9 microM, respectively, for purified recombinant mouse enzyme; the corresponding Vmax values were 2.5, 1.3 and 2.7 micromol x min( 1) x mg(-1) protein, respectively, for human enzyme, and 2.3, 1.2 and 2.2 micromol x min(-1) x mg(-1) protein, respectively, for mouse enzyme. The 5 lipoxygenase-activating-protein inhibitor, MK-886, was active against both human and mouse recombinant leukotriene C4 synthase with IC50 values of 3.1 microM and 2.7 microM respectively. These findings confirm that the leukotriene C4 synthases belong to a gene family that includes the 5-lypoxygenase-activating protein and suggest that the C-terminal domain of leukotriene C4 synthase may not be critical for its conjugation function. PMID- 8706659 TI - Calcium-activated opsin phosphatase activity in retinal rod outer segments. AB - We describe the presence in bovine retinal rod outer segments of a phosphatase which dephosphorylates phosphoopsin with an efficiency similar to that of PP2A, and which is stimulated by submicromolar levels of Ca2+ (half-maximal activation, 0.4-0.5 microM). This enzyme is designated CA2+ -activated opsin phosphatase (CAOP). CAOP has a molecular mass of 70-75 kDa as determined by gel filtration on Superose 12 and exhibits reversible Ca2+ -dependent oligomerization. An unidentified protein of approximately 25 kDa is necessary for full activity of CAOP and for cooperative binding of Ca2+ (h > 2). CAOP does not require Mg2+ and is inhibited by okadaic acid (median inhibitory concentration > 25 microM), which suggests that it is related to the PP1/2A/2b class of protein phosphatases. Like PP2B, CAOP is inhibited by trifluoperazine (median inhibitory concentration 40 microM), but calmodulin has no effect on CAOP activity, and CAOP is inhibited by mastoparan at much higher concentrations than PP2b. This combination of properties suggests that CAOP is not identical to any characterized protein phosphatase. Since the cytoplasmic concentration of Ca2+ -sensitive opsin phosphatase activity suggests that light-dependent Ca2+ levels may control rhodopsin dephosphorylation. PMID- 8706660 TI - The mitochondrial permeability transition pore is modulated by oxidative agents through both pyridine nucleotides and glutathione at two separate sites. AB - We studied the modulation of the permeability transition pore (MTP), a cyclosporin-A-sensitive channel, in deenergized mitochondria. Rat liver mitochondria were incubated in a potassium gluconate medium and treated with uncoupler, valinomycin, oligomycin and A23187. Under these conditions the membrane and Donnan potentials are collapsed, and no ion gradients are maintained, allowing the study of the dependence of MTP opening on the Ca2+ concentration under a variety of oxidative conditions without the complexities arising from changes of the membrane potential and matrix pH, and from secondary ion redistribution. Cross-linking of mitochondrial dithiols with arsenite or phenylarsine oxide, or treatment with tert-butylhydroperoxide leading to complete oxidation of glutathione, increased the sensitivity of MTP opening to Ca2+. This effect could be fully prevented by prior treatment of mitochondria with monobromobimane and restored by reduction with dithiothreitol. The effect of tert butylhydroperoxide was not additive with that of AsO, and it was completely blocked by modification of reduced glutathione with 1-chloro 2,4-dinitrobenzene through glutathione-S-transferase, indicating that oxidized glutathione affects the pore through the AsO-reactive and PhAsO-reactive dithiol. Oxidation of mitochondrial pyridine nucleotides by a variety of treatments also increased the sensitivity of MTP opening to Ca2+ under conditions where glutathione was maintained in the reduced state. This effect could be fully prevented when reduced pyridine nucleotides levels were reestablished with 2-hydroxybutyrate but not by treatment with monobromobimane or dithiothreitol. The effects of dithiol cross-linking or oxidation, and of pyridine nucleotide oxidation on the MTP were additive. The contribution of each of these two oxidative events, when they were induced at the same time, could still be selectively blocked by monobromobimane and dithiothreitol. The effects of dithiol cross-linking or oxidation, and of pyridine nucleotide oxidation on the MTP were additive. The contribution of each of these two oxidative events,when they were induced at the same time, could still be selectively blocked by monobromobimane and dithiothreitol, or by 2 hydroxybutyrate, respectively. Complete oxidation of pyridine nucleotides did not affect the reactivity of the dithiol towards monobromobimane, indicating that it remained in the reduced state. After transient opening of the MTP, sensitivity to pyridine nucleotide oxidation was lost while sensitivity to dithiol cross-linking was retained. These data indicate that the dithiol belongs to an MTP regulatory component which is larger than the MTP cutoff of about 1500 Da, or is non diffusible. Taken together, these findings indicate that the MTP is influenced by oxidation-reduction events at two separate sites that can be distinguished experimentally, and that these sites are not connected by common oxidation reduction intermediates other than glutathione. PMID- 8706661 TI - Molecular cloning and characterization of a transmembrane surface antigen in human cells. AB - The mouse mAb 6C6, raised against a plasma-membrane preparation from human breast cancer cells, reacts with an antigen that appears to be overexpressed in human breast cancers and other human tumors. Here we describe the cDNA cloning and characterization of the antigen recognized by the 6C6 mAb. The isolated cDNA clone encodes a protein of 246 amino acids, with a predicted molecular mass of 27 991 Da. The protein contains three amino-terminal hydrophobic regions, which could represent transmembrane domains, and a hydrophilic carboxy-terminal region, which we show to be extracellular. The identity of the protein encoded by the cloned cDNA as the 6C6 antigen was confirmed by in vitro translation and immunoprecipitation experiments, and by transfection into cell lines that do not react with the 6C6 mAb, which resulted in the expression of a 28-kDa surface protein that was recognized by the antibody. The 6C6 antigen appears to be a type II transmembrane protein, with multiple membrane-spanning domains and a long extracellular non-glycosylated carboxy-terminal domain, to which the 6C6 epitope has been mapped. The overall structure of the protein and weak amino acid similarities with a family of multiple-transmembrane-spanning-domain proteins that includes some antigens (such as L6, CD63/ME491 and CO-029) that are overexpressed in tumor cells, suggest that the 6C6 antigen may belong to this family of proteins. PMID- 8706662 TI - Cell-free conversion of a ubiquitous nuclear protein into a killer-cell-specific form that binds to the NF-P enhancer element of the mouse perforin gene. AB - Two nuclear factors, designated NF-PI and NF-P2, have been shown to bind to an enhancer 9-base motif (5'-ACAGGAAGT-3', NF-P motif) present within the 5' flanking region of the mouse perforin gene. Our previous studies have shown that, although NF-P1 and NF-P2 differ in cell-type distribution and molecular mass, with NF-P2 being killer-cell-specific and smaller, the two factors appear to share common DNA-binding subunit(s). We have postulated that the biochemical event involved in the induction of NF-P2 could be the dissociation of a non-DNA binding subunit from NF-P1, rendering the newly formed NF-P2 transcriptionally active. By using a cell-free system in the present study, we have demonstrated that a variety of chemical agents capable of denaturing or dissociating protein complexes, including guanidinium/HCl, detergents (SDS plus Nonidet P-40) and high salt solutions, could convert NF-P1 into NF-P2. Unlike in intact cells, where induction of NF-P2 is restricted to killer lymphocytes, this conversion occurred in nuclear extracts derived from both cytotoxic lymphocytes and non-cytotoxic cells. Although the mechanism that restricts the induction of NF-P2 to killer- lymphocytes in vivo remains unresolved, these results support the hypothetical 'dissociation' model for the generation of NF-P2. The results also imply that the absence of perforin expression in non-cytotoxic cells may be due to the suppression of the induction of the killer-cell-specific trans-acting factor NF P2. PMID- 8706663 TI - Acceptor substrate specificity of a cloned GD3 synthase that catalyzes the biosynthesis of both GD3 and GD1c/GT1a/GQ1b. AB - To address the role of alpha2,8-sialyltransferase (GD3 synthase) in the biosynthesis of gangliosides, we examined the substrate specificity of the enzyme. In the ganglioside synthesis pathway, it has been generally accepted that sialyltransferase II (SAT II) catalyzes the production of GD3 from GM3, and sialyltransferase V (SAT V) catalyzes the production of GD1c/GT1a/GQ1b from GM1h/GD1a/GT1b. However, acceptor specificity of the clones GD3 synthase that was isolated from human melanoma cells [Nara, K., Watanabe, Y., Maruyama, K., Kasahara, K., Nagai. Y. & Sanai, Y. (1994) Proc. Natl Acad. Sci. USA 91, 7952 7956] has revealed that this enzyme utilized the gangliosides containing the terminal Sia(alpha2-3)Gas structure of the carbohydrate moiety, which includes GM3, GM1b, GD1a and GT1B as exogenous substrates. Kinetic data also showed that the enzyme was able to utilize both GM3 and GM1b/GD1a/GT1b as acceptor substrates. These data indicate that the enzyme catalyzes the formation of not only GD3 but also GD1c, GT1a, and GQ1B in vitro. Furthermore, by transfection of the cloned human alpha2,8-sialyltransferase cDNA, transient and stable expression of GT1a and GQ1b wa also observed in COS-7 cells and Swiss 3T3 cells that originally lacked SAT II and SAT V activities. These observations indicate that the enzyme has both SAT II and SAT V activities in vivo. PMID- 8706664 TI - A recombinant insect-specific alpha-toxin of Buthus occitanus tunetanus scorpion confers protection against homologous mammal toxins. AB - We have constructed a cDNA library from venom glands of the scorpion Buthus occitanus tunetanus and cloned a DNA sequence that encodes an alpha-toxin. This clone was efficiently expressed in Escherichia coli as a fusion protein with two Ig-binding (Z) domains of protein A from Staphylococcus aureus. After CNBr treatment of the fusion protein and HPLC purification, we obtained approximately 1 mg recombinant apha-toxin/l bacterial culture. The toxin, called Bot XIV, displays no toxicity towards mammals but is active towards insects as shown by its paralytic activity against Blatella germanica cockroach and by electrophysiological studies on Periplaneta americana cockroaches. The Bot XIV protein fused to two Z domains is highly immunogenic in mice and induces production of antisera that specifically recognize and neutralize highly toxic components that had been injected into mice. This fusion protein could be very useful for development of potent protective antisera against scorpion venoms. PMID- 8706665 TI - Evidence that anaerobic oxidation of toluene in the denitrifying bacterium Thauera aromatica is initiated by formation of benzylsuccinate from toluene and fumarate. AB - Toluene is degraded anoxically to CO2 by the denitrifying bacterium Thauera aromatica. Toluene first becomes oxidized to benzoyl-CoA by O2-independent reactions. Benzoyl-CoA is then reduced to non-aromatic products by benzoyl-CoA reductase. We set out to study the reactions employed for the initial activation of toluene and its oxidation to the level of benzoate. Evidence is provided for a novel way of toluene degradation based on experiments with cell-free extracts and with whole toluene-grown cells: Cell-free extracts oxidized [14C]toluene to [14C]benzoyl-CoA via several radioactive intermediates. This reaction was strictly dependent on the presence of fumarate, coenzyme A and nitrate as electron acceptor; acetyl-CoA and ATP were not necessary for the reaction. The first product formed in vitro was benzylsuccinate; (2H8)toluene was converted to (2H7)benzylsuccinate. Formation of benzylsuccinate from toluene was independent of coenzyme A and nitrate, but it required the presence of fumarate. Other tricarboxylic acid cycle intermediates were converted to fumarate in cell extracts and therefore could partially substitute for fumarate. [14C]Benzylsuccinate was oxidized further to [14C]benzoyl-CoA and [14C]benzoate in cell extracts if coenzyme A and nitrate were present. No benzyl alcohol and benzaldehyde and no phenylpropionate could be detected as intermediates. In isotope trapping experiments with cell suspensions, two intermediates from [14C]toluene were detected, benzoate and benzylsuccinate. This corroborates the sequence of reactions deduced from in vitro experiments. A hypothetical degradation pathway for the anaerobic oxidation of toluene to benzoyl-CoA via an initial addition of fumarate to the methyl group of toluene and following beta oxidation of the benzylsuccinate formed is suggested. PMID- 8706666 TI - Dephosphorylation of the mannose-6-phosphate recognition marker is localized in later compartments of the endocytic route. Identification of purple acid phosphatase (uteroferrin) as the candidate phosphatase. AB - The mannose-6-phosphate (Man6P) recognition marker in lysosomal proteins is known to be dephosphorylated after the delivery of lysosomal proteins to the endosome/lysosome compartment. The rate of Man6P recognition marker inactivation depends on the cell type and lysosomal protein. In the present study we show that in BHK 21 cells, which rapidly dephosphorylate lysosomal proteins, the recognition marker is stable in the endosomal compartment, to which lysosomal enzymes such as arylsulfatase A are delivered during endocytosis at 20 degrees C. Dephosphorylation depends on the transfer of internalized lysosomal enzymes from the 20 degrees C compartment to later compartments, most likely lysosomes. This transfer is sensitive to NH4C1 and nocodazole. In vitro experiments identified purple acid phosphatase (uteroferrin) as a candidate for the lysosomal phosphatase catalyzing in vivo the dephosphorylation of Man6P recognition marker. PMID- 8706667 TI - The Aspergillus nidulans penicillin-biosynthesis gene aat (penDE) is controlled by a CCAAT-containing DNA element. AB - Analysis of the promoter of the penicillin biosynthesis aat (penDE) gene of Aspergillus nidulans using band-shift assays led to the identification of a CCAAT containing DNA element which was specifically bound by a protein (complex). The identified DNA element was localised about 250 bp upstream of the transcriptional start sites of aat. Substitution of the CCAAT core sequence by GATCC led to a fourfold reduction of expression of an aat-lacZ gene fusion. The identified binding site thus was functional in vivo and positively influenced at expression. Partial purification of the CCAAT binding protein and cross-competition experiments provided evidence that the binding protein is identical to the identified putative penicillin-regulatory protein PENR1, binding to the CCAAT element in the bidirectional intergenic promoter region between acvA (pcbAb) and ipnA (pcbC). Hence, PENR1 seems to be involved in the regulation of all three penicillin-biosynthesis genes. Cross-competition experiments demonstrated that the promoter region of the corresponding aat (penDE) gene of Penicillium chrysogenum was capable to dilute the shift of the A. nidulans probe with PENR1, suggesting the presence of a similar regulatory mechanism in this fungus. Taken together with previous data, CCAAT-containing DNA elements thus seem to represent major cis-acting sites in the promoters of beta-lactam-biosynthesis genes. PMID- 8706668 TI - The copper-topaquinone-phenylhydrazine-adduct geometry in Escherichia coli amine oxidase derivatized with phenylhydrazines substituted with 19F-NMR relaxation measurements. AB - The copper quinoprotein amine oxidase from Escherichia coli was derivatized with phenylhydrazine, substituted with a F3C group at the ortho, meta, or para position. The derivatization of the topaquinone cofactor was verified by ultraviolet/visible spectroscopy. The reduction (with dithionite) of Cu(II) to Cu(I), which was required to obtain reference samples, was verified by EPR spectroscopy. 19F-NMR spectroscopy was carried out on the derivatized enzyme forms, and the spectra showed the line-broadening effect due to the paramagnetic Cu(II). The distance between the Cu and the mean of the three F positions in the F3C groups was calculated by means of the Solomon-Bloembergen equation for the distance-dependent contribution of CU(II) to the transversal-relaxation time of the F resonance. Assuming that the F3C-phenylhydrazines in the enzyme are always aligned towards the Cu in the same way, four configurations can be envisaged that should be taken into account to determine the topology of the two cofactors. Based on these configurations, two spatial positions were found where the calculated distances triangulated, each of these positions having a symmetry related counterpart above or below the topaquinone-phenylhydrazine plane. If it is assumed that the geometric positions of the phenylhydrazine and topaquinone moieties in the adduct remain the same in the derivatized enzymes, a number of minimum distances between the Cu and certain atoms in the topaquinone moiety of the adduct can be calculated (1.52 +/- 0.06 nm from the C2-O, 1.30 +/- 0.04 nm from the C4-O, and 1.26 +/- 0.04 nm from the C5-N). However, one of the configurations yields very similar distances between the Cu and the C2-O and C4 O. Therefore, no conclusions can be made with regard to which OH group is closest to the Cu. By application of the same approach to the 19F-NMR data obtained for porcine-plasma marine oxidase [Williams, T J. & Falk, M.C.(1986) J. Biol. Chem. 261, 15949- 15954] we observed substantial differences between the topologies of the cofactors in the two enzymes. Possible reasons for this are discussed. PMID- 8706670 TI - Evidence for Cu(I) clusters and Zn(II) clusters in neuronal growth-inhibitory factor isolated from bovine brain. AB - Neuronal growth-inhibitory factor (GIF), a central-nervous-system-specific metallothionein-like protein, has been isolated by means of an improved isolation procedure from bovine brain. The native protein contains 4-5 Cu+ and 2-2.5 Zn2+, which results in an overall stoichiometry of 6-7 mol metal ions/mol protein. Native Cu, ZN-GIF and the Zn2+ -substituted and Cd2+-substituted metalloforms have been characterized by means of electronic-absorption, CD, magnetic-circular dichroism (MCD) and low-temperature (77 K) Cu(I)-luminescence spectroscopy. Analysis of the metal-induced-charge-transfer transitions below 300 nm in the electronic-absorption and CD spectra of Cu, ZN-GIF revealed spectral features characteristic of metal-thiolate coordination. The presence of formally spin forbidden 3d --> 4s Cu(I)-cluster-centered transitions, above 300 nm in the corresponding CD and MCD spectra indicate the existence of a Cu(I) cluster. The 77-K luminescence spectrum of Cu, ZN-GIF revealed two emissive bands at approximately 420 nm and 570 nm, which were reported also for CU4 clusters in mammalian Cu8-metallothionein. By analogy with Cu8-metallothionein, we propose the presence of a Cu4 cluster with similar electronic structure in native GIF. However, the determined Cys/Cu+ ratio of approximately 2:1 in Cu, Zn-GIF is higher than the ratio found in mammalian Cu(I)-metallothionein forms (approximately 1.6:1 ), which implies that the coordination geometry of CU+ binding sites is different in the CU4 Cluster. The spectroscopic characterization of Zn2+-substituted and Cd2+-substituted GIF (6-7 metal ions/protein) showed CD and MCD features at positions identical to those reported for the well characterized mammalian Zn7-metallothionein and Cd7-metallothionein. Therefore, it is inferred that the cluster organization in GIF with divalent metal ions is comparable to that found in mammalian metallothioneins. The effect of metal ions on the protein structure with regard to the biological function of GIF is discussed. PMID- 8706669 TI - Cloning and expression of cystolic phospholipase A2 (cPLA2) and a naturally occurring variant. Phosphorylation of Ser505 of recombinant cPLA2 by p42 mitogen activated protein kinase results in an increase in specific activity. AB - Full-length cytosolic phospholipase A2 (cPLA2) was cloned from U937 cells and polymorphonuclear leukocytes (PMNLs) while a naturally occurring variant of cPLA2, which lacks residues Val473-Ala749 but has a C-terminal extension of ILMNLSEYMLWMSKVKRFM (DcPLA2) was cloned from PMNLs and mononuclear leukocytes. We were unable to clone DcPLA2 from U937 cells. When cPLA2 and DcPLA2 were expressed in insect cells, both proteins were detected in cell lysates by SDS/PAGE as single bands of apparent molecular masses 100 kDa and 57 kDa, respectively. Full length cPLA2 was phosphorylated stoichiometrically by p42 mitogen-activated protein (MAP) kinase in vitro at a similar rate to other physiological substrates of this protein kinase and the major site of phosphorylation was identified by amino acid sequencing as Ser505. [32P]Ser(P)505 in cPLA2 was only dephosphorylated at a slow rate by mammalian tissue homogenates. Protein phosphatases 2A, 2B and 2C all contributed significantly to the overall dephosphorylation of cPLA2. The phosphorylation of cPLA2 by p42 MAP kinase correlated with an approximately 1.5-fold increase in specific enzyme activity which was reversed by dephosphorylation. PMID- 8706671 TI - The solution structure of the synthetic circular peptide CGVSRQGKPYC. NMR studies of the folding of a synthetic model for the DNA-binding loop of the ssDNA-binding protein encoded by gene V of phage M13. AB - The cyclic disulfide peptide CGVSRQGKPYC was prepared to obtain a constrained analogue of residues 17-27 of the DNA-binding loop of the gene-V-encoded sDNA binding protein of filamentous bacteriophage M13. Amino acid sequences very similar to that of the beta-loop have been found in various phage-encoded ssDNA binding proteins, and it has been proposed that such a loop may occur as a common motif in this class of proteins. The conformation, in aqueous solution, of the synthetic gene-V-protein binding-loop analogue has been investigated by means of two-dimensional-1H-NMR techniques. Subsequent structure calculations show that the molecule forms a beta-loop that includes a turn formed by three residues. This structure, very unusually for a cyclic disulfide peptide, is highly similar to that of the analogous part of the binding loop of the native protein. Comparison with experiments on other cyclic disulfide peptides indicates that the formation, of the beta-sheet (beta-hairpin) secondary structure is essentially governed by the amino acid composition of the 11-residue sequence. The disulfide bridge in the 11-residue sequence is essential for conformational stability, as indicated by the finding that the open peptide analogue that encompasses residues Ser17-Ser27 does not adopt a detectable secondary structure in water. The bridge replaces the role of the loop formed by residues 49-58 in the protein, which act as a scaffold to hold the N-terminal and C-terminal ends of the DNA-binding loop together. PMID- 8706672 TI - Participation of two Ser-Ser-Phe-Tyr repeats in interleukin-6 (IL-6)-binding sites of the human IL-6 receptor. AB - The alpha-subunit of interleukin-6 (IL-6) receptor is a member of the hematopoietin receptor family. The alignment of its amino acid sequence with those of other members of this family (human somatotropin receptor/murine IL-3 receptor beta and human IL-2 receptor beta) has suggested that amino acids included in two SSFY repeats found in each of its hematopoietin receptor domains, contribute to the binding of the ligand. The involvement of these amino acids in IL-6 binding and signal transduction was studied by site-directed mutagenesis and molecular modelling. We present a computer-derived three-dimensional model of the IL-6/IL-6 receptor complex based on the structure of the human somatotropin/human somatotropin receptor complex. This model allowed the location of distinct regions important for IL-6 and gp130 binding. We show that some of the residues included in the SSFY repeats located in our IL-6 receptor model in the loops between beta-strands E and F of domain-I and B' and C', of domain-II, participate in the formation of a major IL-6-binding site. These residues are necessary for IL-6 and gp130 binding and for signal transduction. Using our IL-6 receptor mutants we mapped the epitopes of our anti-(IL-6 receptor) neutralising monoclonal antibodies to these residues. Our results demonstrate that a generic hematopoietin receptor family structural module can be used for the study of both alpha and beta receptor subunits belonging to this family. PMID- 8706673 TI - Dissimilatory sulfite reductase revisited. The desulfoviridin molecule does contain 20 iron ions, extensively demetallated sirohaem, and an S = 9/2 iron sulfur cluster. AB - Assimilatory sulfite reductase contains a sirohaem that is very weakly coupled to a [4Fe-4S] cubane, i.e. five iron atoms in total. Dissimilatory sulfite reductase is a complex system with 20 Fe atoms/alpha 2 beta 2 gamma 2 hexamer. A recent revision of the purification procedure for the Desulfovibrio vulgaris dissimilatory enzyme has afforded a preparation of only 10 Fe atoms hexamer, this has led to the convulsion that the topology of prosthetic groups parallels that of the assimilatory system [Wolfe, B. M., Lui, S. M. & Cowan, J. A. (1994) Eur. J. Biochem. 223, 79-89]. The new purification procedure has been reproduced but the claimed molecular properties are not reproducible. The highly purified, active desulfoviridin contains 20, not 10, Fe atoms/molecule: the sirohaem is extensively dematallated, not metallated; and the S = 9/2 iron-sulfur cluster is present, not absent. PMID- 8706674 TI - Proteases from Trypanosoma brucei brucei. Purification, characterisation and interactions with host regulatory molecules. AB - African trypanosomes contain proteases that may be released into the bloodstream of their infected hosts. This paper describes a novel, combined isolation of a cysteine proteinase (called trypanopain-Tb) and a serine oligopeptidase (which we call oligopeptidase-Tb) from Trypanosoma brucei brucei, as well as a comparison of the activities of these two enzymes against several host regulatory molecules. The enzymes differed in various respects. Firstly, purified trypanopain-Tb was shown to readily cleave proteins such as gelatin maximally at acidic pH. In contrast, oligopeptidase-Tb, which is optimally active at alkaline pH, did not hydrolyse proteins larger than 4 kDa. However, it readily hydrolysed various polypeptides, including neurotensin and atrial natriuretic factor. The interaction of the two enzymes with mammalian protease inhibitors also differed. Cystatins and alpha2-macroglobulin effectively inhibited trypanopain-Tb, with the Ki values for cystatin C and low-molecular-mass kininogen (approximately 10(-11) M) predicting, that trypanopain-Tb is likely to be effectively controlled by these inhibitors if released into the host bloodstream. In contrast, oligopeptidase-Tb was not inhibited by serpins or (a2-macroglobulin, suggesting that it may remain active if released into the host bloodstream. In support of these in vitro results, the blood of trypanosome-infected rats displayed no trypanopain-Tb-like activity, but exhibited high oligopeptidase-Tb-like activity. Thus, while trypanopain-Tb seems likely to be confined to an intracellular role within the parasite, oligopeptidase-Tb has the potential to remain active in the host bloodstream and so contribute directly to pathogenesis. PMID- 8706675 TI - Electropermeabilization of intact maize cells induces an oxidative stress. AB - By applying electric field pulses through cell suspensions, cell membranes can be permeabilized transiently, giving free access to the cytosol. Electropulsation is now routinely used in cell biology when introducing various molecules such as proteins and nucleic acids into the cell. But the molecular and cellular bases of cell electropermeabilization are still unclear. In the present study, we observed that electropermeabilization of intact black Mexican sweet (BMS) maize cells induces a generation of oxygen species (oxidative jump). Using the chemiluminescent probe lucigenin, we have shown that the electro-induced chemiluminescent response depends on the level of the stress factor as shown by its dependence on the electric parameters (electric field intensity, duration, and number of pulses). While the electroinduced cell permeabilization has a short life, the oxidative jump that is triggered by this electropermeabilization is a much longer-lived response. The electroinduced loss in viability is linearly correlated to permeabilization. However, there is no correlation between the oxidative jump and the loss in viability. The modulation of oxygen species electroinduction by antioxidant products (dimethylsulfoxide, sodium L-ascorbate, and glutathione) does not lead to an increase in cell viability. Such results are different to those observed with mammalian cells and indicate that even if the same phenomenon is observed with mammalian cells and indicate that even if the same phenomenon is observed when pulsing mammalian or intact plant cells, the associated metabolic response is not the same. PMID- 8706676 TI - Structure of the gene encoding the sea urchin blastula protease 10 (BP10), a member of the astacin family of Zn2+-metalloproteases. AB - Blastula protease 10 (BP10), a metalloprotease of the astacin family, is secreted at the blastula stage by the sea urchin embryo. The BP10 gene shows a precise temporal and spatial regulation during embryogenesis. It has been cloned from a sea urchin lambda genomic library and the transcription unit has been entirely sequenced. It spans 6kb and contains seven exons (2.8 kb) and six introns (3.2 kb). Sequence comparison and phylogeny analysis show that BP10 belongs to a sub family of molecular proteins which all play a role during development. In the two cases where the exon/intron organization of the gene is known (BP10 and tolloid), the modular structure of the protein is not reflected at the gene level, which indicates that this sub-family probably did not evolve by exon shuffling. PMID- 8706677 TI - Transcription of four satellite DNA subfamilies in Diprion pini (Hymenoptera, Symphyta, Diprionidae). AB - Four satellite DNA subfamilies Ps, Pv, Pv65 and Ec, resulting from the evolution of a common ancestral motif, were isolated and characterized in the genomic DNA of Diprion pini, a phytophagous of Pinus sylvestris. Consensus sequences were 148 312 bp long. Sequence analyses revealed that these satellite subfamilies have evolved from a 45-bp ancestral motif. The amounts of each satellite in the genome (0 - 10%) and the accessibility of the DNA to restriction enzymes were sex dependent. The migration of each monomer in polyacrylamide gels and the electrophoretic migration of d(AT) n > or = 3 residues showed that all four satellite subfamilies are curved. Their transcription was analyzed using reverse transcription and the polymerase chain reaction. Three satellite DNA subfamilies were transcribed on both strands, and in both sexes. However, the female satellite DNAs seem to be more actively transcribed than those of males, indicating that transcription is not constitutive. The lack of any significant open reading frame in satellite monomers indicates that the RNA may function as structural or catalytic RNA rather than encoding protein. PMID- 8706678 TI - Substitution of fifty four homologue (Ffh) in Escherichia coli with the mammalian 54-kDa protein of signal-recognition particle. AB - The fifty four homologue (Ffh) of Escherichia coli promotes the translocation of a subset of periplasmic, membrane and secreted proteins across the cytoplasmic membrane. The ffh gene product is essential for cell viability and efficient protein export. Here we show that the mammalian homologue signal-recognition particle (SRP) 54 kDa is not able to suppress the translocation defect in an Ffh conditional mutant Wam 113 [Phillips, G.J. & Silhavy, T.J. (1992) Nature 359, 744 746]. The expression of SRP 54kDa, which is increased when Ffh is suppressed in the Wam 113 strain, causes a pleiotropic defect characterised by cell elongation, and increased accumulation of precursor proteins. The accumulation of precursors of outer membrane protein A (Omp A) and maltose-binding protein (MBP), See-B dependent pre-proteins, was less than the Ffh-dependent proteins ribose-binding protein (RBP) and beta-lactamase. Sec B expression was suppressed by Ffh expression. The recombinant SRP 54 kDa, which forms a ribonucleoprotein complex in E coli, was shown to bind to precursor proteins, but is unable to interact with the filamentous temperature-sensitive Y (Fts Y) membrane receptor of the translocation machinery. PMID- 8706679 TI - Purification and structural characterization of bovine cathelicidins, precursors of antimicrobial peptides. AB - Cathelicidins are a novel family of antimicrobial peptide precursors from mammalian myeloid cells. They are characterized by a conserved N-terminal region while the C-terminal antimicrobial domain can vary considerably in both primary sequence and length. Four cathelicidins, proBac5, proBac7, prododecapeptide and proBMAP-28, have been concurrently purified from bovine neutrophils, using simple and rapid methodologies. The correlation of ES-MS data from the purified proteins with their cDNA-deduced sequences has revealed several common features of their primary sequence, such as the presence of N-terminal 5-oxoproline (pyroglutamate) residues and two disulfide bridges in a 1-2, 3-4 arrangement. The N-terminal domains of the cathelicidins present one or two Asp-Pro bonds, which are particularly acid-labile in proBac5 and proBac7, but stable in prododecapeptide. This suggests that the spatial organization around these bonds may vary in different cathelicidins, and favour hydrolysis in some cases. An unexpected feature of the prododecapeptide is that it exists as dimers formed by three possible combinations of its two isoforms. The isolation of a truncated, monomeric form of this protein, lacking the cysteine-containing antimicrobial dodecapeptide, indicates that dimerization occurs via disulfide bridge formation at the level of the C-terminal domain and that the dodecapeptide is likely released as a dimer from its precursor. Sequence-based secondary structure predictions and CD results indicate for cathelicidins a 30-50% content of extended conformation and <20% content of alpha-helical conformation, with the alpha-helical segment placed near the N-terminus. Finally, similarity searching and topology-based structure prediction underline a significant sequential and structural similarity between the conserved N-terminal domain of cathelicidins and cystatin-like domains, placing this family within the cystatin superfamily. When assayed against cathepsin L, unlike the potent cystatin inhibitors, three of the four cathelicidins show only a poor inhibitory activity (Ki = 0.6-3 microM). PMID- 8706680 TI - Characterization of the interaction between annexin I and profilin. AB - Annexin I belongs to a family of calcium-dependent phospholipid-binding and membrane-binding proteins. Although many of the biochemical properties and the three-dimensional structure of this protein are known, its true physiological roles have yet to be thoroughly defined. Its putative functions include participation in the regulation of actin microfilaments dynamics, proposed after the discovery of an interaction with actin. In accordance with this hypothesis, we found that annexin I can also interact with profilin. We used different methods, overlay and surface plasmon resonance (BIAcore), to measure the parameters of the association equilibrium, i.e. k(on), k(off) and k(d). The affinity of annexin I for profilin was between 10(7) M and 10(8) M. High concentrations of KCl did not prevent the interaction, although a slight decrease in affinity was observed. Calcium, a modulator of annexin I functions interfered only marginally with the association, in a manner comparable to magnesium. Proteins or compounds known to interact with annexin I or profilin were found to inhibit the annexin-I--profilin interaction when added in the reaction medium. Recombinant profilin exhibited a slightly lower affinity than natural platelet protein when measured with BIAcore. Due to the submembrane localisation of annexin I and the regulatory activity of profilin on the cytoskeleton, an interaction between annexin I and profilin may therefore be implicated in the regulation of some cellular functions, particularly those governing membrane cytoskeleton dynamic organization. PMID- 8706681 TI - Experimental procedure for a hydrogen peroxide assay based on the peroxidase oxidase reaction. AB - A low-cost assay method that is able to measure H2O2 concentrations as low as the nano-molar range is described. The assay solution contains NADH, horseradish peroxidase, and superoxide dismutase at PH 7.5. After the addition of the sample, the decrease in NADH concentration measured by spectrophotometry is proportional to the H2O2 concentration. Because of superoxide dismutation, a high amplification factor defined as moles NADH oxidised per mole H2O2 added is obtained, which allows the sensitivity limit of the method to be greatly improved. We have established the conditions under which the amplification factor can be stabilised at a high level: the best compromise is to increase both the horseradish peroxidase and superoxide dismutase concentrations. Finally, we have also shown that coupled to specific oxidases, our assay method is suitable for measuring very low concentrations of biochemicals that can be oxidized by oxygen with H2O2 production. PMID- 8706682 TI - The role of a beta barrel loop 4 extension in modulating the physical and functional properties of long-chain 2-hydroxy-acid oxidase isozymes. AB - Peroxisomal long-chain 2-hydroxy-acid oxidase, an FMN-dependent enzyme, catalyzes the oxidation of a variety of L-2-hydroxy acids into keto acids at the expense of oxygen. We recently reported the cloning and sequencing of its CDNA and the existence of a weakly expressed isozyme [Belmouden, A., Le, K. H. D., Lederer, F. & Garchon, H. J. (1993) Eur. J. Biochem. 214, 17-251. This isozyme, beta 2 differs from the major one in having a three-residue insertion, -VRK-, in loop 4 of the beta 8 alpha 8 barrel. In the crystal structures of homologous flavocytochrome beta 2, and glycolate oxidase, the corresponding region of loop 4 is disordered. We now report on the constitutive high-level expression of isozymes beta 1, and beta 2 in Escherichia coli under control of the lambda pL promoter, and on the influence of the E. coli genetic background and the growth medium on the expression level. We describe the properties of isozyme beta 2 and compare them with those of pure isoform beta 1. The visible spectra of the purified enzymes differ in the position of the near-ultraviolet band of the prosthetic group. pH titration studies indicate that the FMN ionizes at N3 at a lower pH than free flavin and that there is a small pKa difference between the isozymes. To our knowledge, the only other known case of a lowered pKa for the protein-bound flavin is that of glycolate oxidase. In the CD spectra of the FMN region, a marked difference between isozymes in the 270-300-nm region appears to be related to the pKa difference for the N3-H bond. Kinetic parameters for a number of substrates and inhibitors are indistinguishable within the limits of experimental error, with the exception of values for kcat for mandelate (the most active substrate), Km for hydroxyhippurate (a new substrate), Ki for cinnamate and oxalate, and Kd for sulfite. The differences are no larger than twofold. The foregoing comparison between isozymes beta 1 and beta 2 shows that the naturally engineered insertion in loop 4 exerts some influence on the flavin spectral properties and the active-site reactivity. Since the corresponding loop 4 regions in the three-dimensional structures of flavocytochrome 2 and glycolate oxidase are 1.5-2.0 nm removed from the flavin, it would appear either that loop 4 has a very different conformation in hydroxy-acid oxidase, or that it may interact with the active site due to mobility. PMID- 8706683 TI - Regulation of prothymosin alpha during the cell cycle. AB - A number of studies have indicated that the small nuclear acidic protein prothymosin alpha is associated with cellular-proliferation events. For example, c-myc causes immediate transcriptional activation of prothymosin alpha, and prothymosin alpha antisense oligonucleotides inhibit myeloma cell division. To investigate the regulation of prothymosin alpha, we examined its mRNA and protein levels during the cell cycle of mononuclear cells and fibroblastic cells. We isolated immunoreactive material from cellular extracts and immunolocalized the protein to the nucleus during the cell cycle. We reported here that the material present in the cells is prothymosin alpha rather than the amino-terminal peptide thymosin alpha 1. [3H]Thymidine-incorporation studies associate maximum accumulation of mRNA and protein with the S/G2 phase of the cell cycle. This induction of prothymosin alpha mRNA seems to resemble cyclin B expression and is more pronounced in fibroblasts. Moreover, transient-transfection experiments indicate that transcription factor E2F is a strong positive regulator of the prothymosin alpha gene. Our results are consistent with the hypothesis that prothymosin alpha is involved in proliferation checkpoints of the cell cycle. PMID- 8706684 TI - Sarco(endo)plasmic reticulum Ca2+ pump and metabolic enzyme expression in rabbit fast-type and slow-type denervated skeletal muscles. A time course study. AB - Recent reports by d'Albis et al. have shown that denervation of 8-day-old rabbit fast-twitch muscle (gastrocnemius) leads to the transformation of the muscle towards a slow phenotype but the changes towards slow-type myosin isoforms and contractile properties of the muscle were temporally uncoordinated. We analyzed the time course of the effects of denervation of the gastrocnemius on the expression of the sarcoplasmic reticulum calcium pump isoforms (SERCA) and on the metabolic state of the muscle. Northern-blot analysis showed a rapid loss of the fast Ca2+ pump isoform (SERCA 1) mRNA from the denervated gastrocnemius which became of the oxidative type. The changes observed were complete as early as 35 days post-natal, i.e at the time when changes in contractile properties were previously observed. Denervation of the slow-twitch soleus led to a 50% decrease in the level of the slow Ca2+ pump isoform (SERCA 2) mRNA and was without effect on the metabolic state of the muscle. These findings extend previous results suggesting that in rabbit, continuous innervation is required for differentiation of fast-twitch muscles but is not an absolute requirement for differentiation of the slow-twitch muscle. PMID- 8706685 TI - The complete cDNA sequence of laminin alpha 4 and its relationship to the other human laminin alpha chains. AB - We previously localised the gene (LAMA4) encoding a novel laminin alpha 4 chain to chromosome 6q21. In this study, we describe the complete coding sequence and compare the protein with the other three known human laminin alpha chains. Although closely linked to LAMA2, the LAMA4 product most closely resembles laminin alpha 3, a constituent of laminin 5. Like laminin alpha 3A, the alpha 4 chain is a truncated version of the alpha 1 and alpha 2 chains, with a much reduced short arm. While the alpha 4 molecule is most similar to alpha 3, it shares some features of the C-terminal domains G4 and G5 in common with alpha 2. Unlike the LAMA3 gene, LAMA4 appears to encode only a single transcript, as determined by 5' rapid amplification of cDNA ends. The cDNA sequence encodes 1816 amino acids, which include a 24-residue signal peptide. The gene is expressed in skin, placenta, heart, lung, skeletal muscle, and pancreas. We have also shown that the mRNA can be readily reverse transcribed and amplified from cultured dermal fibroblasts. PMID- 8706686 TI - Molecular characterization of a saline-soluble lectin from a parasitic fungus. Extensive sequence similarities between fungal lectins. AB - It has been proposed that the interactions between several parasitic and pathogenic fungi and their hosts are mediated by soluble lectins present in the fungus. We have cloned and analyzed a gene encoding such a lectin (AOL) from the nematophagous fungus Arthrobotrys oligospora (deuteromycete). The deduced primary structure of the AOL gene displayed an extensive similarity (identity 46.3%) to that of a gene encoding a lectin (ABL) recently isolated from the mushroom Agaricus bisporus (basidiomycete), but not to any other fungal, microbial, plant or animal lectins. The similarities between AOL and ABL were further demonstrated by the observation that an antibody specific for AOL cross-reacted with ABL. Together with data showing that AOL has a binding specificity that is similar to that of ABL [Rosen, S., Bergstrom, J., Karlsson, K.-A. & Tunlid, A. (1996) Eur. J. Biochem. 238, 830-837], these results indicate that AOL and ABL are members of a novel family of saline soluble lectins present in fungi. Southern blots indicated that there is only one AOL gene in the genome encoding a subunit (monomer) of the lectin. The primary structure of AOL did not show the presence of a typical N-terminal signal sequence. Comparison of the deduced primary structure with the molecular mass of AOL as determined by electrospray mass spectrometry (16153 Da), indicated that AOL has an acetylated N-terminal but no other post-translational modifications, and that a minor isoform is formed by deamidation. Circular dichroism (CD) spectroscopy suggested that the secondary structure of AOL contains 34% beta-sheets, 21% alpha-helix, and 45% turns and coils. PMID- 8706687 TI - A multispecific saline-soluble lectin from the parasitic fungus Arthrobotrys oligospora. Similarities in the binding specificities compared with a lectin from the mushroom agaricus bisporus. AB - Several fungi can express high levels of saline-soluble and low-molecular-mass lectins that bind to glycoproteins such as fetuin and different mucins but not bind to any monosaccharides. In this paper, we report the binding specificities of such a lectin (designated AOL) isolated from the nematophagous fungus Arthrobotrys oligospora. The results show that AOL is a multispecific lectin that interacts with the following ligands: (a) Several sulfated glycoconjugates including sulfatide, dextran sulfate, and fucoidan. The specificity of this binding was indicated by experiments showing that none of the tested neutral- and sialic-acid-containing glycolipids, chondroitin sulfates B and C, heparin, and polyvinyl sulfate bound to AOL; (b) Phosphatidic acid and phospatidylglycerol, two out of several tested phospholipids. (c) N-linked and O-linked sugar chains bound to intact fetuin. The involvement of such sugar structures was demonstrated by analyzing the binding of AOL to chemically deglycosylated (trifluoromethanesulfonic acid) fetuin. Treating fetuin with O-glycosidase and N glycosidase indicated that AOL bound to Gal beta GaLNAc alpha-Ser/Thr and to some N-linked complex sugars, respectively. Further assays demonstrated that AOL could interact with several other glycoproteins containing O-linked and/or N-linked sugar chains. The observations that AOL did not bind to free N-linked sugars isolated from fetuin, or to fetuin treated with trypsin or pronase, or to any of the tested neoglycoproteins and glycolipids with neutral- or sialic acid containing sugars, indicated that the sugar chains need to be bound to an intact peptide backbone to interact with AOL. We have recently shown that the deduced primary structure of AOL has a high similarity to the sequence of a saline soluble lectin isolated from the mushroom Agaricus bisporus (ABL) (Rosen, S., Kata, M., Persson, Y., Lipniunas, P. H., Wikstrom, M., van den Hondel, C. A. M. J. J., van den Brink, J. M., Rask, L., Heden L.-O. and Tunlid, A., see companion paper). It is well known that ABL binds to Gal beta 3GaLNAc alpha-Ser/Thr, and in this paper we demonstrate that ABL binds to sulfatide, phosphatidic acid, phospatidylglycerol, and possibly also to the same N-linked complex sugars as AOL. The above data indicate that AOL and ABL are members of a novel family of fungal lectins sharing similar primary structure and binding properties. PMID- 8706688 TI - A selenium-containing phospholipid-hydroperoxide glutathione peroxidase in Schistosoma mansoni. AB - The 100000Xg supernatant parasite platyhelminth Schistosoma mansoni exhibits a glutathione peroxidase activity with the substrate phosphatidylcholine hydroperoxide. Purification yielded a protein of 20 kDa molecular mass both on gel filtration column chromatography and SDS/PAGE, thus suggesting that S. mansoni expresses a protein similar to the mammalian selenoenzynic phospholipid hydroperoxide glutathione peroxidase. Kinetic analysis and substrate specificity corroborated this assumption, the second-order rate constants for the oxidation of the ground-state enzyme (k+1) being higher with phosphatidylcholine hydroperoxide than with other peroxide substrates, such as cumene liydroperoxide or H2O2, and quantitatively similar to those of mammalian phospholipid hydroperoxide glutathione peroxidase. Partial sequencing of the protein and selenium measurement by neutron activation analysis established that the purified peroxidase corresponded to the product of the S. mansoni gene previously reported and supposed to encode a selenium-containing glutathione peroxidase [Roche, C., Williams, D. L., Khalife, J., LePresle, T., Capron, A. & Pierce, R. J. (1994) Cloning and characterization of gene encoding Schistosoma mansoni glutathione peroxidase, Gene 138, 149 - 152]. S. mansoni thus contains a scienoperoxidase sharing molecular mass, catalytic efficiency and substrate specificity with phospholipid-hydroperoxide glutathione peroxidase, dismantling the concept that those enzymes are unique to vertebrate organisms. PMID- 8706689 TI - Peroxisomal and mitochondrial carnitine acetyltransferases of the n-alkane assimilating yeast Candida Tropicalis. Analysis of gene structure and translation products. AB - A genomic DNA clone encoding carnitine acetyltransferases (EC 2.3.1.7), localized in two subcellular organelles, peroxisomes and mitochondria of an n-alkane assimilating yeast Candida tropicalis, was isolated from the yeast lambda EMBL library using a carnitine acetyltransferase CDNA probe. Nucleotide sequence analysis disclosed that the open reading frame was 1881 bp, corresponding to 627 amino acids with a molecular mass of 70760 Da. Comparison of the predicted amino acid sequence of the C. tropicalis enzyme with that of Saccharomyces cerevisiae mitochondrial matrix carnitine acetyltransferase revealed 46.3% identity. It was noticeable that the C. tropicalis enzymes had amino acid sequences similar to both proposed mitochondrial and peroxisomal targeting signals. When the C. tropicalis gene was expressed in S. cerevisiae using its own 5'-upstream region, a 12-fold increase in activity was observed. Western blot analysis revealed the presence of two major proteins whose sizes corresponded to the peroxisomal and mitochondrial proteins detected in C. tropicalis. This suggested that peroxisomal and mitochondrial carnitine acetyltransferases were encoded by one gene, as suggested for the S. cerevisiae enzyme. Furthermore, we have separated and purified these enzymes from peroxisomes and mitochondria of C. tropicalis, and analyzed the amino-terminal amino acid sequences of each. The amino-terminal sequence of the mitochondrial enzyme suggested that a signal sequence had been cleaved during translocation into mitochondria. Concerning the peroxisomal enzyme, the evidence obtained indicated that in vivo the translation was initiated at the second methionine of the open reading frame. PMID- 8706690 TI - Human N-acetylglucosaminyltransferase III gene is transcribed from multiple promoters. AB - We have isolated cDNA clones for the human N-acetylglucosaminyltransferase III (GlcNAc-transferase III) gene. Two of them, H15 and H20, contain 5' non-coding regions that are totally different from each other except for 8 bp adjacent to the putative initiation codon. Analysis of one of the genomic cosmid clones containing the GlcNAc-transferase III coding region, Hug3, revealed the 5' non coding regions of H15 and H20 contain two and one exons, respectively, in addition to the exon containing the coding region (exon 1). These have arisen as the result of alternative splicing. The transcription-initiation sites were determined by primer-extension analysis and 5'-rapid amplification of cDNA ends (RACe). Both H15-specific and H20-specific primers gave cDNAs longer than those expected from the lengths of H15 and H20, and a primer complementary to the region around the intron/exon junction near the putative initiation codon also gave distinct signals. Promoter activities of the 5'-flanking regions of H15, H20 and exon 1 were measured in a human hepatoblastoma cell line, HuH-6 cells by luciferase assays. The 5'-flanking region of exon 1 was the most active, whilst that of H15 was several times less active, and that of H20 was inactive. Our study suggests that multiple promoters of the GlcNAc-transferase III gene contribute to the complex regulation of this gene. PMID- 8706691 TI - Characterization of xanthine dehydrogenase from the anaerobic bacterium Veillonella atypica and identification of a molybdopterin-cytosine-dinucleotide containing molybdenum cofactor. AB - The molybdenum-containing iron-sulfur flavoprotein xanthine dehydrogenase from the anaerobic bacterium Veillonella atypica has been purified approximately 800 fold with a yield of approximately 40% and a specific activity of approximately 70 micromol ferricyanide reduced x min(-1) x mg protein(-1) with xanthine as electron donor, which corresponds to approximately 30 micromol xanthine oxidized x min(-1) x mg protein(-1) with methylene blue as electron acceptor. The 129-kDa enzyme was a non-covalent heterotrimer with large (82.4 kDa), medium (28.5 kDa) and small (18.4 kDa) subunits. The N-termini of the small and medium polypeptides of V. atypica xanthine dehydrogenase and the corresponding domains of eukaryotic xanthine dehydrogenases were similar, whereas the N-terminus of the large polypeptide was unrelated to eukaryotic xanthine dehydrogenases. The enzyme contained 0.86 atoms Mo, 1.75 atoms Fe, 1.61 atoms acid-labile sulfur and 0.68 molecules FAD/molecule, which corresponds to a 1:2.0:1.9:0.8 molar ratio. Acid hydrolysis revealed 0.95 mol CMP and 0.80 mol AMP/mol xanthine dehydrogenase. After treatment of the enzyme with iodoacetamide, di(carboxamidomethyl)molybdopterin cytosine dinucleotide was identified, which indicates that molybdopterin cytosine dinucleotide is the organic portion of the V. atypica xanthine dehydrogenase molybdenum cofactor. The enzyme and its molybdenum cofactor occurred in a 1:1 molar ratio. Xanthine dehydrogenases from eukaryotic sources are characterized by a domain structure and the presence of duplicate copies of two types of [2Fe-2S) clusters. In contrast, the xanthine dehydrogenase from V. atypica had a heterotrimeric subunit structure and a single [2Fe-2S] cluster. In addition, the enzyme indicates the presence of a molybdopterin dinucleotide as a constituent of a xanthine dehydrogenase molybdenum cofactor. PMID- 8706692 TI - A human peroxisome-proliferator-activated receptor-gamma is activated by inducers of adipogenesis, including thiazolidinedione drugs. AB - We have cloned a human cognate of the mouse peroxisome-proliferator-activated receptor-gamma (hPPAR gamma) from a human placenta cDNA library. Sequence analysis reveals a high degree of similarity with the mouse receptor and, like other PPAR, hPPAR gamma forms heterodimers with the retinoid X receptor alpha (RXR alpha) and binds in vitro to DNA elements containing direct repeats of the sequence TGACCT. In common with mouse PPAR gamma, hPPAR gamma is expressed strongly in adipose tissue, but significant levels also are detectable in placenta, lung and ovary. In vitro trans-activation data suggest hPPAR gamma is only poorly activated by xenobiotic peroxisome proliferators, although certain fatty acids and eicosanoids are potent activators of this receptor. Both mouse and human PPAR gamma are capable of being activated by thiazolidinedione drugs, although the two receptors appear to differ in their sensitivity to these compounds. Taken together, these data suggest a high degree of structural and functional similarity between mouse and human PPAR gamma, and provide evidence for variation in human receptor structure which may result in differential sensitivity to activators. PMID- 8706693 TI - Inhibition by gangliosides of Bacillus cereus phospholipase C activity against monolayers, micelles and bilayer vesicles. AB - The effect of complex glycosphingolipids (gangliosides) on the activity of phospholipase C from Bacillus cereus was studied using lipid monolayers, mixed micelles and small unilamellar vesicles containing phosphatidylcholine as substrate. In all artificial membrane systems assayed, gangliosides exhibit qualitatively similar inhibitory properties. Gangliosides decrease the enzyme activity irrespective of the aggregation structure in which the substrate is offered to B. cereus phospholipase C, and they do not affect the adsorption process of the enzyme. The modulatory effect of gangliosides occurs at the level of the interface, affecting both the maximum rate of catalysis of the enzyme already adsorbed and the availability of the substrate in a suitable organization for enzyme catalysis to take place. PMID- 8706694 TI - Expression of tyrosine-sulfated secretory proteins in Xenopus laevis oocytes. Differential export of constitutive and regulated proteins. AB - Xenopus laevis oocytes were used to study the tyrosine sulfation and secretion of exogenous proteins. Secretogranin II (SgII), a tyrosine-sulfated protein found in secretory granules of a wide variety of endocrine cells and neurons, became tyrosine-sulfated by the oocytes when expressed by injection of poly(A)-rich RNA isolated from the neuroendocrine cell line PC12. The same result was observed when SgII was expressed from cloned SgII cRNA, showing that its tyrosine sulfation did not require the coexpression of exogenous tyrosylprotein sulfotransferase (TPST) but occurred by means of the endogenous oocyte TPST. Sulfophilin, an artificial protein consisting of 12 repeats of a heptapeptide tyrosine-sulfation site, was highly sulfated upon injection of its RNA, indicating the presence of TPST levels sufficient for stoichiometric sulfation of appropriate reporter proteins. Comparison of the secretion of [35S]sulfate labelled SgII with that of sulfophilin and an exogenous heparan sulfate proteoglycan (HSPG), two proteins delivered to the cell surface by the constitutive pathway of secretion, revealed striking differences. The majority of sulfophilin and the HSPG was found in the medium, whereas that of SgII was found intracellularly. Prolactin, another secretory granule protein, showed the same secretion behaviour as SgII. These results show that oocytes express TPST and that these cells secrete constitutive and regulated secretory proteins in a differential manner. PMID- 8706695 TI - Dietary modulation of the mRNA stability of trypsin isozymes and the two forms of secretory trypsin inhibitor in the rat pancreas. AB - The stability of the mRNAs encoding pancreatic trypsin isozymes, namely the cationic form and the two anionic forms I and II, as well as that of the secretory trypsin inhibitors I and II, were studied in rats fed on either a high protein diet, or a protein-free diet compared with a standard diet for a 10-day period. Either immediately or 3 h and 6 h after injecting the transcription inhibitor, actinomycin D, the mRNA levels were quantified by performing dot-blot hybridization with specific oligonucleotide probes. Under high-protein dietary conditions, the stability of the mRNAs coding for anionic trypsin II and cationic trypsin showed no change, whereas that of anionic trypsin I and the two forms of secretory trypsin inhibitor were affected. The mRNA half-life of anionic trypsin I and trypsin inhibitor II increased, in sharp contrast with that of trypsin inhibitor I, which decreased. When rats were fed on a protein-free diet, the stabilities of both anionic trypsin forms and trypsin inhibitor I increased, whereas that of trypsin inhibitor II decreased and that of cationic trypsin remained unchanged. The present results show the existence of differences in the mechanisms whereby gene expression of trypsin isozymes and secretory trypsin inhibitors is regulated, although they are synthesized in parallel in the pancreatic acinar cell and stored in zymogen granules before being secreted into the intestinal lumen. PMID- 8706696 TI - N,N'-Bisformyl dityrosine is an in vivo precursor of the yeast ascospore wall. AB - The amino acid, dityrosine, is a major component of the spore wall surface of the yeast Saccharomyces cerevisiae, where it is part of a highly cross-linked macromolecular network of yet unknown chemical structure, consisting mostly of glucosamine, dityrosine and few other amino acids. Biosynthesis of the dityrosine moiety of this network consists of several steps, including the chemical modification of free L-tyrosine and the subsequent oxidative cross-linking of the modified tyrosine residues (catalyzed by a cytochrome P-450), leading to soluble dityrosine-containing spore wall precursors. We isolated, purified and characterized the dityrosine-containing precursor that appears late in spore wall synthesis and that is thought to be directly incorporated into the maturing spore wall. Chemical and spectroscopic analyses showed that this precursor is N,N' bisformyl dityrosine. In addition, we identified a tyrosine-containing spore wall precursor as N-formyl tyrosine. The elucidation of the chemical structure of soluble spore wall precursors is crucial for the characterization of the function of the enzymes involved in maturation of the spore surface, e.g. by in vitro systems. A dityrosine-containing fragment, which was solubilized from mature spore walls by partial hydrolysis, was identified as N-formyl dityrosine. Mature spore walls contain significant amounts of N-formyl dityrosine and N,N'-bisformyl dityrosine. This supports the assumption that the dityrosine-containing macromolecular network on the spore surface has an unusual, non-peptidic structure. PMID- 8706697 TI - Cloning and sequencing of human gp330, a Ca(2+)-binding receptor with potential intracellular signaling properties. AB - We present here the complete primary structure of human gp330, the human variant of the principal kidney autoantigen causing Heymann membranous glomerulonephritis in rats. The deduced 4655 amino acid residues give a calculated molecular mass of 519636 Da for the mature protein and consists of a probable 25-amino-acid N terminal signal peptide sequence, an extracellular region of 4398 amino acids, a single transmembrane-spanning domain of 23 amino acids, and an intracellular C terminal region of 209 amino acid residues. Three types of cysteine-rich repeats characteristic of the low density lipoprotein receptor (LDLR) superfamily are present in human gp330. In the extracellular region, there are a total of 36 LDLR ligand-binding repeats, comprising four distinct domains, 16 growth factor repeats separated by eight YWTD spacer regions, and one epidermal growth factor like repeat. No consensus cleavage sequence for the processing endoprotease furin is detected in human gp330. The intracellular tail contains not only two copies of the F(X)NPXY coated-pit mediated internalization signal characteristic of LDLR superfamily members, but also intriguing and potentially functional motifs including several Src-homology 3 recognition motifs, one Src-homology 2 recognition motif for the p85 regulatory subunit of phosphatidylinositol 3 kinase, and additional sites for protein kinase C, casein kinase II and cAMP /cGMP-dependent protein kinase. There is approximately 77% amino acid identity between human and rat gp330 with minor differences between the extracellular and intracellular regions. Recently gp330 has been implicated in Ca2+ regulation in the parathyroid, the placenta, and the renal tubule, but its overall physiological and pathological role still remains uncertain. PMID- 8706698 TI - A site-directed mutagenesis study of Saccharomyces cerevisiae pyrophosphatase. Functional conservation of the active site of soluble inorganic pyrophosphatases. AB - We report the expression and initial characterization of 19 active-site variants of Saccharomyces cerevisiae inorganic pyrophosphatase (PPase), including measurements of thermostability, oligomeric structure and specific activity at pH 7.2. 13 of the 19 conservative substitutions resulted in at least a fivefold decrease in activity, indicating that these residues are important for yeast PPase catalysis. The E58D, D117E, D120E and D152E variants had no activity under the conditions tested, suggesting that Glu58, Asp117, Asp120 and Asp152 may have crucial roles in catalysis. The effects of the mutations on catalytic activity were very similar to those observed with the corresponding variants of Escherichia coli PPase, proving conclusively that the active site and mechanism of soluble PPases are conserved. The D71E variant was more thermostable and the K56R, R78K, D115E and K154R variants were more thermolabile than the wild-type enzyme, whereas subunit:subunit interactions were somewhat weakened by the K56R, R78K, Y89F and K154R substitutions. These results suggest that Lys56, Asp71, Arg78, Tyr89, Asp115 and Lys154 are structurally important for yeast PPase. PMID- 8706699 TI - Characterization of the human small-ribosomal-subunit proteins by N-terminal and internal sequencing, and mass spectrometry. AB - Reverse-phase HPLC was used to fractionate 40S ribosomal proteins from human placenta. Application of a C4 reverse-phase column allowed us to obtain 27 well resolved peaks. The protein composition of each chromatographic fraction was established by two-dimensional polyacrylamide gel electrophoresis and N-terminal sequencing. N-terminally blocked proteins were cleaved with endoproteinase Lys-C, and suitable peptides were sequenced. All sequences were compared with those of ribosomal proteins available from data bases. This allowed us to identify all proteins from the 40S human ribosomal subunit in the HPLC elution profile. By matrix-assisted laser-desorption ionization mass spectrometry the masses of the 40S proteins were determined and checked for the presence of post-translational modifications. For several proteins differences to the deduced sequences and the calculated masses were found to be due to post-translational modifications. PMID- 8706700 TI - Purification and characterisation of an alpha-glucan phosphorylase from the thermophilic bacterium Thermus thermophilus. AB - An alpha-glucan phosphorylase has been purified 4500-fold from the thermophilic bacteria Thermus thermophilus. In contrast to other bacterial phosphorylases the thermophilic enzyme seems neither to be inducible by maltose nor repressed by glucose. T. thermophilus phosphorylase shares major properties with known mesophilic phosphorylases such as pyridoxal 5'-phosphate content (1 M pyridoxal P/M subunit), subunit molecular mass (about 90 kDa) and inhibitor constants. The optimum temperature of T. thermophilus phosphorylase was observed at 70 degrees C in the pH range 5.5-6.5. While at 25 degrees C the subunit composition of the thermophilic enzyme is an octameric form, the preferential form at the optimum temperature of 70 degrees C seems to be a dimer. Most remarkably, in the direction of synthesis and degradation the limiting size of the oligosaccharide substrate is shorter by one glucose residue than the minimum size of substrate degraded by other alpha-glucan phosphorylases. Maltotetraose and glycogen are degraded with rates similar to that observed with maltoheptaose (Vmax = 18 U/mg). Correspondingly, maltotriose functions as primer in the synthesis direction. Differences in fluorescence and absorption spectra of the cofactor and the failure of arsenate acting as a substrate indicate that the active site structure of T. thermophilus phosphorylase differs from that of known alpha-glucan phosphorylases. PMID- 8706701 TI - Interaction of the lantibiotic nisin with membranes revealed by fluorescence quenching of an introduced tryptophan. AB - Nisin is a lantibiotic produced by strains of Lactococcus lactis subsp. lactis. The target for nisin action is the cytoplasmic membrane of gram-positive bacteria. To aid understanding of its mode of action, the interaction of nisin with vesicles of differing phospholipid composition were investigated by fluorescence techniques, using a variant of nisin in which the isoleucine at position 30 was replaced by a tryptophan residue. Activity of the site-directed variant containing tryptophan was established to be similar to that of the wild type peptide. Fluorescence experiments showed a blue shift of the emission wavelength maximum in the presence of lipid vesicles, indicating that the tryptophan residue enters a more hydrophobic environment. Quenching experiments with aqueous and membrane-restricted quenchers (iodide and spin-labelled lipids, respectively) both confirmed a non-aqueous environment for the Trp30 residue, and implied that the residue resides between 0.36 nm and 0.52 nm from the centre of the membrane, depending on the lipid identity. The results clearly demonstrate that nisin interacts strongly with the hydrophobic phase of lipid vesicles. This interaction is stronger in the presence of negatively charged lipids suggesting their importance in the functional interaction of nisin with membranes. PMID- 8706702 TI - The osmotic properties and free energy of formation of the actomyosin rigor complexes from rabbit muscle. AB - We have studied the osmotic properties of the calcium-regulated actomyosin complexes from skeletal muscle at the protein osmotic pressure of 18 kPa and a different actin-to-myosin molar ratios. Essentially, protein solutions were equilibrated against a solution of poly(ethylene glycol) 40,000 of known macromolecular osmotic pressure. At the end of the equilibration the water and the protein masses of the protein solutions were determined gravimetrically and the protein molar concentration was calculated. In this reconstructed system we have found following, at the actin-to-molar ratio of 2.6 (the most likely stoichiometry of these two proteins in the dense region of the A band) the average distance between the myosin filaments is 34.2 nm, this equals the interfilament distance in the intact fibre of muscle in rigor, at the sarcomere length of 3.38 micrograms. The formation of the F-actin-myosin and of the tropomyosin-F-actin-myosin rigor complexes involves the largest free energy changes, -5.38 kJ/mol myosin and -5.67 kJ/mol myosin, respectively. The formation of the troponin-tropomyosin-F-actin-myosin(Ca) rigor complex from myosin and troponin-tropomyosin-F-actin(Ca) occurs with the free energy change of -3.43 kJ/mol myosin. Of these -3.43 kJ, -1.81 kJ are provided by the endergonic conversion of troponin-tropomyosin-F-actin(EGTA) into troponin-tropomyosin-F actin (Ca). The transition of myosin and of troponin-tropomyosin-F-actin(EGTA) into the -F-actin-myosin(Ca) rigor complex is accompanied by a 5.8% increase of volume. The increase of volume is due to a large influx of water, which is essentially protein-hydration water. PMID- 8706703 TI - Different effects of insulin and platelet-derived growth factor on phosphatidylinositol 3-kinase at the subcellular level in 3T3-L1 adipocytes. A possible explanation for their specific effects on glucose transport. AB - Insulin stimulates glucose uptake by induction of the translocation of vesicles that contain the glucose transporter Glut 4 to the plasma membrane. Phosphatidylinositol 3-kinase (PtdIns 3-kinase), which is thought to be involved in intracellular trafficking, could play a critical role in insulin-induced glucose transport. In 3T3-L1 adipocytes, insulin and platelet-derived-growth factor (PDGF) stimulated glucose uptake by 5.8-fold and 2.4-fold, respectively, but PDGF had no significant effect on Glut 4 translocation. Nevertheless, both hormones activated PtdIns 3-kinase activity in total cell extracts. However, insulin and PDGF had different effects on the stimulation of PtdIns 3-kinase activity in several subcellular fractions, and the movements of insulin-receptor substrate (IRS) 1 and the p85 subunit of PtdIns 3-kinase between subcellular compartments. PDGF stimulated PtdIns 3-kinase activity almost exclusively in the plasma membrane, and induced translocation of the p85 subunit from the cytosol to the plasma membrane, where the PDGF receptor was phosphorylated on tyrosine residues. In contrast, insulin stimulated PtdIns 3-kinase activity in the plasma membrane, in low-density microsomes (LDM) and in cytosol. Furthermore, insulin induced the translocation of p85 from the cytosol to LDM and the translocation of IRS 1 from LDM to the cytosol. These data indicate that insulin and PDGF have different effects on the activation of PtdIns 3-kinase and on the movement of IRS 1 and PtdIns 3-kinase between subcellular compartments. We would like to suggest that a crucial event in the stimulation of glucose uptake by insulin could be that insulin, but not PDGF, induces activation of PtdIns 3-kinase in the cytosol and in LDM, the compartment enriched in Glut-4-containing vesicles. PMID- 8706704 TI - Cloning, sequencing and sites of expression of genes for the hydroxyarginine containing adhesive-plaque protein of the mussel Mytilus galloprovincialis. AB - A segment of Mytilus galloprovincialis foot protein 3 (Mgfp-3) cDNA was amplified by means of reverse-transcription (RT)/PCR with degenerate primers. The 5' and 3' regions of the cloned segment were amplified by means of rapid amplification of cDNA ends. The 5'-region clones had almost identical nucleotide sequences, but two sequences were found among 3'-region clones. The Mgfp-3 coding region was amplified between a 5' untranslated sequence and one of two 3' untranslated sequences. Two cDNA clones which encoded variants Mgfp-3A and Mgfp-3B, were isolated. These two clones encoded proteins with 70 and 77 amino acid residues, of which the first 24 residues are predicted to be signal peptides. The existence of additional variants was suggested by the sequences of other clones. Thus, it was suggested that Mgfp-3 genes constitute a gene family. RT/PCR of RNA from developing larvae indicates that Mgfp-3 genes are transcribed after settlement. RT/PCR of RNA from major organs and in situ hybridization of the foot indicate that Mgfp-3 genes are transcribed in a limited part of the foot, i.e., the phenol gland and a distal part of the accessory gland. PMID- 8706705 TI - Stimulation of ADP-ribosyl cyclase activity of the cell surface antigen CD38 by zinc ions resulting from inhibition of its NAD+ glycohydrolase activity. AB - The lymphocyte cell surface antigen, CD38, which has an amino acid sequence similar to Aplysia ADP-ribosyl cyclase, catalyzes not only the hydrolysis of NAD+ and 1-(5-phospho-beta-D-ribosyl) adenosine 5'-phosphate cyclic anhydride (cyclic ADP-ribose) but also the formation of cyclic ADP-ribose from NAD+. To characterize the bifunctional enzyme properties, we produced the recombinant CD38 fused with a maltose-binding protein (MBP-CD38). Zinc ions stimulated the ADP ribosyl cyclase activity of MBP-CD38, but inversely inhibited its NAD+ glycohydrolase activity which was approximately 100-fold dominant to the cyclase activity in the absence of Zn2+. Such dual effects of Zn2+ were also observed in the native membrane-bound CD38 of HL-60 cells which had been caused to differentiate by retinoic acid. Zinc ions inhibited the NAD+ glycohydrolase reaction catalyzed by MBP-CD38 in an uncompetitive manner, whereas they enhanced the ADP-ribosyl cyclase reaction without affecting the Km value for NAD+. There was an increase in the fluorescence intensity of a hydrophobic fluorescent probe, 8-anilino-1-naphthalenesulfonate, in the presence of MBP-CD38. The fluorescence increase was further enhanced by the addition of Zn2+ with a shift in the maximum emission wavelength from 484 nm to 470 nm, suggesting that Zn2+ caused conformational changes of MBP-CD38. These results indicate that Zn2+ directly interacts with CD38 to stimulate its ADP-ribosyl cyclase with inhibition of its NAD+ glycohydrolase, probably due to prevention of the access of water molecule to an intermediate of the enzymesubstrate complex. PMID- 8706706 TI - Characterization of a ribosomal inhibitory polypeptide of protein phosphatase-1 from rat liver. AB - About 4% of the spontaneous phosphorylase phosphatase activity in a rat liver extract was associated with the ribosomal fraction and stemmed from both protein phosphatase-1 (PP-1) and protein phosphatase-2A (PP-2A). However, after repeated washing, only PP-1 remained bound to the ribosomes. The activity of ribosome associated PP-1 (PP-1R) was partially latent and could be increased 2-3-fold by incubation with trypsin and an additional 50% by incubation with low concentrations of exogenous type-1 catalytic subunit. In contrast, incubation of the ribosomal fraction with MgATP resulted in a 50% drop in the activity of PP 1R. We have purified from a ribosomal extract a basic polypeptide (pI > or = 10.5) of 23 kDa that potently inhibited PP-1. This ribosomal inhibitor of PP-1, termed RIPP-1, was at least 30-times less efficient in inhibiting other major Ser/Thr protein phosphatases (PP-2A, PP-2B and PP-2C). RIPP-1 was identified as a non-competitive inhibitor of PP-1 with a substrate-dependent potency. The lowest Ki (approximately 20 nM) was obtained with phosphorylase and myelin basic protein as substrates. Besides instantaneously inhibiting the type-1 catalytic subunit, RIPP-1 also converted the catalytic subunit in a time-dependent manner (t 1/2 = 45 min at 25 degrees C) into a less active conformation. Unlike the inhibition, this slow inactivation was not reversed by the removal of RIPP-1. We propose that RIPP-1 accounts, at least in part, for the latency of PP-1R. PMID- 8706707 TI - Crystal structure of flavodoxin from Desulfovibrio desulfuricans ATCC 27774 in two oxidation states. AB - The crystal structures of the flavodoxin from Desulfovibrio desulfuricans ATCC 27774 have been determined and refined for both oxidized and semi-reduced forms to final crystallographic R-factors of 17.9% (0.8-0.205-nm resolution) and 19.4% (0.8-0.215-nm resolution) respectively. Native flavodoxin crystals were grown from ammonium sulfate with cell constants a = b = 9.59 nm, c=3.37nm (oxidized crystals) and they belong to space group P3(2)21. Semireduced crystals showed some changes in cell dimensions: a = b = 9.51 nm, c=3.35 nm. The three dimensional structures are similar to other known flavodoxins and deviations are found essentially in the isoalloxazine ring environment. Conformational changes are observed between both redox states and a flip of the Gly61-Met62 peptide bond occurs upon one-electron reduction of the FMN group. These changes influence the redox potential of the oxidized/semiquinone couple. Modulation of the redox potentials is known to be related to the association constant of the FMN group to the protein. The flavodoxin from D. desulfuricans now studied has a large span between E2 (oxidized --> semiquinone) and E1 (semiquinone --> hydroquinone) redox potentials, both these values being substantially more positive within known flavodoxins. A comparison of their FMN environment was made in both oxidation states in order to correlate functional and structural differences. PMID- 8706708 TI - Conservation of aconitase residues revealed by multiple sequence analysis. Implications for structure/function relationships. AB - Aconitases have recently regained much attention, because one member of this family, iron regulatory protein-1 (IRP-1), has been found to play a dual role as a cytoplasmic aconitase and a regulatory RNA-binding protein. This finding has highlighted a novel role for Fe-S clusters as post-translational regulatory switches. We have aligned 28 members of the Fe-S isomerase family, identified highly conserved amino acid residues, and integrated this information with data on the crystallographic structure of mammalian mitochondrial aconitase. We propose structural and/or functional roles for the previously unrecognized conserved residues. Our findings illustrate the value of detailed protein sequence analysis when high-resolution crystallographic data are already available. PMID- 8706709 TI - Catalytic activity of complex I in cell lines that possess replacement mutations in the ND genes in Leber's hereditary optic neuropathy. AB - Short-chain ubiquinone analogues act as electron acceptors and as inhibitors in the lymphoblast mitochondria of ND1/3460 mutants, which indicates structural changes in the ubiquinone-binding domain of Complex I in this mutant. The ND4/11778 mutant and two secondary ND5 mutants studied are associated with reductions of at least 50, 35 and 30% in the catalytic rate constant, respectively. However, the efficiency of oxidative phosphorylation is unaffected in all these ND mutants. The rate of respiration is only slightly limited by Complex I in lymphoblast mitochondria. Consequently, there is a 30-35% reduction in the electron flow through Complex I compared with that through Complex II, and an increased lactate/pyruvate ratio, in the ND1 and ND4 mutants, but these factors were unaffected in the secondary ND5 mutants. Energy metabolism is thus less severely affected in the secondary mutants than in the primary mutants, which supports the division into these two categories. An increased ubiquinone-10 content in the mitochondrial membrane of all the mutants, and enhanced succinate dehydrogenase and citrate synthase activities in the ND4 mutant, are proposed to be compensatory changes. The efficiency of these changes and the level of kinetic limitation of respiration by Complex I in each tissue are proposed to determine the clinical development of the disease. PMID- 8706710 TI - Nucleoside-diphosphate kinase from Streptomyces coelicolor. AB - Nucleoside-diphosphate (NDP) kinase was purified from crude extracts of Streptomyces coelicolor to over 90% homogeneity in a single step using an adenosine 3',5'-cyclic monophosphate (cAMP) binding column. The specific activity of protein in the fraction eluted with cAMP (400 U/mg) was about 3600-fold higher than that in the crude extract. This enzyme was autophosphorylated in the presence of [y-32P]ATP. The high-energy phosphoenzyme intermediate was stable in alkali and highly labile in acid; this suggests the presence of an N-phosphate amino acid (most probably a histidine residue). A tetrameric form of the 15-kDa protein was suggested by its apparent molecular mass (66 kDa) on a gel filtration column. The measured Michaelis constant (Km) for ATP was 85 microM. The IC50 for cAMP of 6 mM suggested weak competitive inhibition. However, no evidence that cAMP acts as an allosteric effector was obtained. The ndk gene from S. coelicolor was isolated and sequenced. The deduced amino acid sequence was very similar to other NDP kinases. However, unique characteristics were also noted, including a truncated C-terminus that makes it one of the smallest NDP kinases reported in the literature. PMID- 8706711 TI - A deletion in the first cysteine-rich repeat of the low-density-lipoprotein receptor leads to the formation of multiple misfolded isomers. AB - The ligand-binding domain of the low-density-lipoprotein (LDL) receptor comprises seven cysteine-rich repeats, each approximately 40 amino acids long. The deletion of two amino acids (Asp26 and Gly27) from the first of these repeats (LB1), leads to a defective LDL receptor, and the clinical syndrome of familial hypercholesterolemia [Leitersdorf, E., Hobbs, H. H., Fourie, A. M., Jacobs, M., van der Westhuyzen, D.R. & Coetzee, G.A. (1988) Proc. Natl Acad. Sci. USA 85, 7912-7916]. Receptors which reach the cell surface fail to bind IgG-C7, a conformation-specific monoclonal antibody directed to LB1. To determine the effects of the two-amino-acid deletion on the folding of the LB1 of the LDL receptor, we have expressed LB1 and the mutant repeat, des-Asp26, Gly27-LB1, as recombinant (rLB1 and des-Asp26, Gly27-rLB1) peptides, and have determined their ability to fold in vitro. Unlike rLB1, which folded into a single isomer that was recognized by IgG-C7 and had three disulfide bonds, des-Asp26, Gly27-rLB1 folded into an equilibrium mixture of four isomers. Each of these isomers contained three disulfide bonds, but none were recognized by IgG-C7. We suggest that LDL receptors in the endoplasmic reticulum (ER) of the cell also fold into an equilibrium mixture of distinct receptor molecules, each with an abnormally folded isomer of des-Asp26, Gly27-LB1, and that the retarded transport of receptors to the cell surface arises because only a subset of the isomers reaches the cell surface. PMID- 8706712 TI - X-ray structure of yeast inorganic pyrophosphatase complexed with manganese and phosphate. AB - The three-dimensional structure of the manganese-phosphate complex of inorganic pyrophosphatase from Saccharomyces cerevisiae has been refined to an R factor of 19.0% at 2.4-A resolution. X-ray data were collected from a single crystal using an imaging plate scanner and synchrotron radiation. There is one dimeric molecule in the asymmetric unit. The upper estimate of the root-mean-square coordinate error is 0.4 A using either the delta A plot or the superposition of the two crystallographically independent subunits. The good agreement between the coordinates of the two subunits, which were not subjected to non-crystallographic symmetry restraints, provides independent validation of the structure analysis. The active site in each subunit contains four manganese ions and two phosphates. The manganese ions are coordinated by the side chains of aspartate and glutamate residues. The phosphate groups, which were identified on the basis of their local stereochemistry, interact either directly or via water molecules with manganese ions and lysine, arginine, and tyrosine side chains. The phosphates are bridged by two of the manganese ions. The outer phosphate is exposed to solvent. The inner phosphate is surrounded by all four manganese ions. The ion-binding sites are related to the order of binding previously established from kinetic studies. A hypothesis for the transition state of the catalytic reaction is put forward. PMID- 8706713 TI - Kinetics of phosphorolysis of 3-(beta-D-ribofuranosyl)adenine and 3-(beta-D ribofuranosyl)hypoxanthine, non-conventional substrates of purine-nucleoside phosphorylase. AB - The properties of two non-conventional substrates of the calf-spleen and Escherichia coli purine nucleoside phosphorylases (PNP), 3-(beta-D ribofuranosyl)adenine (RibfAde) and 3-(beta-D-ribofuranosyl)hypoxanthine (RibfHyp), are described. In contrast to Ado, RibfAde is a substrate for the mammalian enzyme. With the calf enzyme, the pseudo-first-order rate constants (Vmax/K(m)) for phosphorolysis of RibfAde and RibfHyp are 3% and 13%, respectively, that for phosphorolysis of Ino, while for E. coli PNP the corresponding values are 22% and 30%, respectively. The Michaelis constants (K(m)) for RibfAde were 800 microM (calf PNP) and 150 microM (E. coli PNP). For RibfHyp, the corresponding K(m) values were 220 microM and 260 microM. Two well characterized inhibitors of calf spleen PNP [9-(2-fluoro-3,4 dihydroxybutyl)guanine] and E. coli PNP (formycin A) were found to inhibit phosphorolysis of RibfAde and RibfHyp with the same inhibition constants as for Ino. Moreover, the inhibition was competitive, which indicates that phosphorolysis of 3-beta-nucleosides occurs at the same active site as for the natural substrate Ino. In particular, the substrate properties of both 3-beta nucleosides are consistent with their binding to the enzyme in the conformation anti to the imidazole ring about the glycosidic bond, which is superimposable on the structure of natural 9-beta-nucleosides in the conformation anti to the pyrimidine ring. The results are examined in relation to present concepts regarding the binding of substrates and inhibitors at the active site(s) of these enzymes. PMID- 8706714 TI - The cooperation between two silencers creates an enhancer element that controls both the lens-preferred and the differentiation stage-specific expression of the rat beta B2-crystallin gene. AB - The rat beta B2-crystallin gene is active only during a specific stage of the differentiation of rat lens fibre cells directed by basic fibroblast growth factor. The regulatory elements that determine the transient activity of this gene are located in the -750/-123 region and in the first intron. Singly, these elements act as silencers, together they constitute an enhancer that is active only during the specific differentiation stage. An additional silencer is found between -123 and -77. The proximal promoter region contains a Pax-6 binding site at -65/-51. In vitro, binding to this site could be detected but, according to in vivo footprinting experiments, this site is not occupied in the endogenous gene. Furthermore, co-expression of Pax-6 did not enhance promoter activity. Finally, mutation or deletion of this site did not affect promoter activity: the region 37/+10 sufficed for basal promoter activity. The cooperation between the -750/ 123 region and the first intron of the beta B2-crystallin gene not only determines the differentiation stage-specific activity of the gene, but also contributes to the highly increased expression in lens cells compared with non lens cells. PMID- 8706715 TI - On the mechanism of selenium tolerance in selenium-accumulating plants. Purification and characterization of a specific selenocysteine methyltransferase from cultured cells of Astragalus bisculatus. AB - Selected members of the genus Astragalus (Fabaceae) are known for their ability to accumulate high levels of selenium, mainly in the form of Se-methyl selenocysteine. With the aid of cell cultures we have investigated the molecular basis for selenium tolerance of these plants. It is shown that cultured cells from a selenium-accumulating Astragalus species synthesize Se-methyl selenocysteine in contrast to those of a non-accumulating species and do not unspecifically incorporate selenium into proteins. The purification and biochemical characterization of a selenocysteine methyltransferase from cultured Astragalus bisculatus cells is described, which does not accept cysteine as a substrate. We propose that this enzyme plays a crucial role in conferring selenium tolerance. PMID- 8706716 TI - Xylose-DNA: comparison of the thermodynamic stability of oligo(2' deoxyxylonucleotide) and oligo(2'-deoxyribonucleotide) duplexes. AB - Measurements of differential scanning calorimetry, ultraviolet absorption and circular dichroism have been performed on two synthetic oligo(2' deoxyxylonucleotides): (A) d[xA)3-(xT)3-(xA)3-(xT)3-T] and (B) d[(xA-xT)6-T], and on the oligo(2'-deoxyribonucleotide) (C) d[(A)3-(T)3-(A)3-(T)3]. Oligonucleotides having 2'-deoxyxylose instead of 2'-deoxyribose exhibit unusual thermodynamic, optical and structural features. At identical concentrations the transition temperatures of the oligo(2'-deoxyxylooligomers) are higher than those of the oligo(2'-deoxyribooligomers) indicating higher stability. The calorimetric transition enthalpy of (C) is 270 +/- 15 kJ . mol-1, the corresponding van't Hoff value is 280 +/- 15 kJ . (mol of cooperative unit)-1. The ratio of delta HvH/delta Hcal = 1.04 suggests all-or-none behaviour for the transition of the 2' deoxyribose oligonucleotide. The analogous parameters of (A) are: delta Hcal = 310 +/- 30 kJ . mol-1, delta HvH = 220 +/- 30 kJ.(mol of cooperative unit)-1. The ratio of 0.71 indicates multistate melting for this compound. The sequence dependence of the thermodynamic quantities becomes apparent when the parameters of the alternating oligo(2'-deoxyxylonucleotide) d[(xA-xT)6-T] are compared to those of d[(xA)3-(xT)3-(xA)3-(xT)3-T). The values are delta Hcal = 330 +/- 30 kJ.mol-1; delta HvH = 180 +/- 15 kJ.(mol of cooperative unit)-1 delta HvH/ delta Hcal = 0.55. The transition enthalpy of the alternating oligo(2' deoxyxylonucleotide) (B) is the highest but the cooperativity of transition is the lowest of the oligonucleotides studied. The circular dichroic spectra of the two oligo(2'-deoxyxylonucleotides) show unusual features in that d[(xA)3-(xT)3 (XA)3-(xT)3-T] exhibits a spectrum that is suggestive of a left-handed double helix, while the spectrum of the alternating oligo(2'-deoxyxylonucleotide) (B) resembles neither that of (C) nor that of (A). PMID- 8706717 TI - Cloning, sequencing and functional overexpression of the Streptococcus equisimilis H46A gapC gene encoding a glyceraldehyde-3-phosphate dehydrogenase that also functions as a plasmin(ogen)-binding protein. Purification and biochemical characterization of the protein. AB - We previously identified DNA sequences involved in the function of the complex promoter of the streptokinase gene from Streptococcus equisimilis H46A, a human serogroup C strain known to express this gene at a high level. As a prerequisite to understanding possible mechanisms that control the balance between the plasminogen activating and plasmin(ogen) binding capacities of H46A, we describe here its gapC gene encoding glyceraldehyde-3-phosphate dehydrogenase (GraP-DH, EC 1.2.1.12), a glycolytic enzyme apparently transported to the cell surface where it functions as a plasmin(ogen).binding protein. The gapC gene was cloned and sequenced and found to code for a 336-amino-acid polypeptide (approximately 35.9 kDa) exhibiting 94.9% sequence identity to the Plr protein from Streptococcus pyogenes shown by others to be capable of plasmin binding [Lottenberg, R., Broder, C. C., Boyle, M. D., Kain, S. J., Schroeder, B. L. & Curtiss, R. III (1992) J. Bacteriol. 174, 5204-5210]. To study the properties of the GapC protein, its gene was inducibly overexpressed in Escherichia coli from QIAexpress expression plasmids to yield the authentic GapC or (His)6GapC carrying a hexahistidyl N-terminus to permit affinity purification. Both proteins were functionally active, exhibiting specific GraP-DH activities of about 80 kat/mol (approximately 130 U/mg) after purification. Their binding parameters [association (ka) and dissociation (kd) rate constants, and equlibrium dissociation constants (Kd = kd/ka)] for the interaction with human Gluplasminogen and plasmin were determined by real-time biospecific interaction analysis using the Pharmacia BIAcore instrument. For comparative purposes, the commercial GraP-DH from Bacillus stearothermophilus (BstGraP-DH), a nonpathogenic organism, was included in these experiments. The Kd values for binding of plasminogen to GapC, (His)6GapC and BstGraP-DH were 220 nM, 260 nM and 520 nM, respectively, as compared to 25 nM, 17 nM and 98 nM, respectively, for the binding to plasmin. These data show that both the zymogen and active enzyme possess low-affinity binding sites for the gapC gene product and that the hexahistidyl terminus does not affect its function. Prior limited treatment with plasmin enhanced the subsequent plasminogen binding capacity of all three GraP DHs, presumably by the exposure of new C-terminal lysine residues for binding to the zymogen. PMID- 8706719 TI - X-ray absorption spectroscopy study of native and phenylphosphorodiamidate inhibited Bacillus pasteurii urease. AB - X-ray absorption spectroscopy (XAS) has been applied to urease from Bacillus pasteurii, a highly ureolytic soil bacterium, with the aim of elucidating the structural details of the nickel-containing active site. The results indicate the presence of octahedrally coordinated Ni2+, in a sphere of six N/O donors at an average distance of 0.203 nm. An average of two histidine residues are bound to nickel. The experimental evidence suggests direct binding of the urease inhibitor phenylphosphorodiamidate to Ni2+. These spectroscopic results are in agreement with previous findings on both plant and microbial ureases, but differ in some respect from the results obtained by X-ray crystallography analysis of Klebsiella aerogenes urease. PMID- 8706718 TI - Follitropin action on the transferrin gene in Sertoli cells is mediated by cAMP responsive-element-binding-protein and antagonized by chicken ovalbumin-upstream promoter-transcription factor. AB - The transcription of the transferrin (Tf) gene is induced by follitropin via cAMP in rat Sertoli cells. We previously demonstrated that the cAMP-responsive-element binding protein (CREB) interacts on the proximal region II (PRII) of the human Tf promoter (Suire et al., 1995). The PRII region is identified as essential for cAMP inducibility of the Tf promoter and contains a CCAAT box. This unexpected result led us to study the relation that exists between CREB and the PRII site. In the liver, CCAAT/enhancer-binding (C/EBP) proteins act at the PRII site. Although these factors are absent in Sertoli cells, their overexpression in Sertoli cells disturbs basal and induced transcription. C/EBP alpha and delta were able to stimulate the basal transcription driven by the -100 to +39 region, placed upstream of the chloramphenicol acetyltransferase (CAT) gene. However, only C/EBP alpha allowed the cAMP-inducible expression. The Ka of CREB bZIP (254 327), a deleted form of CREB, for the CRE site (3.92 x 10(8)M-1) and for the PRII site (1.38 x 10(8)M-1) were determined using the surface plasmon resonance (SPR) method. The Ka values were similar, although the derived kinetics were different: higher ka and kd of CREB for the PRII site were found compared with the CRE site. Since we observed important dissociation kinetics, we hypothesized that the binding of CREB to the PRII site is stabilized by CREB-binding protein (CBP) or by chicken-ovalbumin-upstream-promoter transcription factor (COUP-TF) binding to PRI site near to PRII. However, we observed that the overexpression of CBP in Sertoli cells did not potentiate the basal and cAMP-stimulated activity of CREB of the -100 to +39Tf-CAT construct. In basal and cAMP-stimulated conditions, COUP TF appeared to repress the transcription driven by the -100 to +39 region in a specific manner. These results demonstrate a direct action of CREB on hTf promoter, which is antagonized by COUP-TF and may explain the transcriptional regulation of Tf by follitropin, via cAMP. PMID- 8706720 TI - Primary structure of omega-hordothionin, a member of a novel family of thionins from barley endosperm, and its inhibition of protein synthesis in eukaryotic and prokaryotic cell-free systems. AB - A new sulfur-rich basic polypeptide, so called omega-hordothionin, has been isolated from barley endosperm by extractions with NaCl and ammonium bicarbonate followed by reverse-phase high performance liquid chromatography. Purified omega hordothionin was found to be homogeneous by SDS/polyacrylamide gel electrophoresis, N-terminal amino-acid sequencing and electrospray-ionization mass spectrometric analysis. The complete primary structure of omega-hordothionin was determined by automatic degradation of the intact molecule and peptides obtained by proteolytic cleavage. Omega-hordothionin consists of a single polypeptide chain of 48 amino acids with a molecular mass of 5508 Da deduced from its amino acid sequence, which fully coincides with the 5508.2 Da determined by electrospray-ionization mass spectrometry. The isolated polypeptide showed a characteristic composition with a high content of basic amino acids (five arginine residues, two lysine residues and six histidine residues) and eight cysteine residues, and has strong sequence identity (66%) with the sorghum SI alpha 1 alpha-amylase inhibitor. Omega-hordothionin, like gamma-hordothionin, exhibited translation inhibitory activity on both eukaryotic cell-free systems from mammalian (rat liver and rabbit reticulocyte lysates) and prokaryotic cell free systems (Escherichia coli). However, in contrast to gamma-hordothionin, omega-hordothionin did not inhibit plant systems such as Triticum aestivum, Cucumis sativus, Vicia sativa and Hordeum vulgare. Gamma-hordothionin also inhibited the alpha-amylase activity from human saliva, while omega-hordothionin and the other different genetic variants of thionins, alpha-hordothionin and beta hordothionin, failed to show any inhibitory effect. PMID- 8706721 TI - Branched synthetic constructs that mimic the physico-chemical properties of apolipoprotein AI in reconstituted high-density lipoproteins. AB - Amphipathic helical repeats are considered as the structural units of numerous apolipoproteins and have been described as being responsible for the interaction of apolipoproteins with phospholipids in high-density lipoproteins (HDL). Furthermore, apolipoproteins, and especially apolipoprotein AI (apoAI), are involved in various biological functions of these circulating particles in plasma. Studies with synthetic peptides corresponding to domains of the apoAI sequence have however shown that short 39-residue fragments do not interact strongly enough with phospholipids to generate particles that correctly mimic the physico-chemical properties of HDL reconstituted with native apoAI [Vanloo, B., Demoor, L., Boutillon, C., Lins, L., Baert, J., Fruchart, J. C., Tartar, A. & Rosseneu, M. (1995) Association of synthetic peptide fragments of human apolipoprotein A-I with phospholipids, J. Lipid Res. 36, 1686-1696.]. Here we show that synthetic branched multimeric peptides, often used as carriers for the design of synthetic vaccines (multiple-antigen peptides), can be used to mimic the physiochemical properties of apoAI in HDL. This type of molecule is obtained by using a small core matrix of Lys residues bearing radially branched synthetic peptides as dendritic arms. We compared the lipid-binding capacities and the structural properties of a linear peptide corresponding to residues 145-183 of apoAI [apoAI-(145-183)-peptide] with those of two multimeric peptides consisting respectively of three [trimeric apoAI-(145-183)] and four copies [tetrameric apoAI-(145-183)] of the selected sequence, branched on a covalent core matrix. This paper provides evidence for the increased abilities of the multimeric peptides to associate with phospholipids compared with the short linear peptides. Moreover, the trimeric apoAI-(145-183) peptide was most efficient in mimicking the physico-chemical and structural properties of native apoAI in reconstituted HDL. As tools adequate to unravel the structure/function relationship of separate apolipoprotein domains are still missing, these multimeric peptides might constitute an alternative approach to linear peptides which are poor mimetics and to protein mutants which are difficult to produce and only provide information about the total sequence. PMID- 8706722 TI - Transcriptional regulation of the lutropin/human choriogonadotropin receptor and three enzymes of steroidogenesis by growth factors in cultured pig Leydig cells. AB - Recent data have shown that Leydig-cell-specific functions, and therefore steroidogenic capacity, can be regulated by lutropin/human choriogonadotropin collectively termed gonadotropin and by several growth factors that are produced by and act within the testis. However, the molecular mechanisms by which these factors regulate Leydig cells are not understood. In the present study, we have investigated the effects of basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), insulin-like growth factor I (IGF-I) and transforming growth factor beta (TGF-beta) on mRNA for the gonadotropin receptor and three steroidogenic enzymes: cytochrome P-450scc, cytochrome P-450 17 alpha hydroxylase/C17-20 lyase (17 alpha-hydroxylase), and 3 beta-hydroxysteroid dehydrogenase. IGF-1, which can enhance testosterone production, increased gonadotropin-receptor density after an increase in receptor mRNA levels, and it increased the level of mRNA for cytochrome P-450scc and 17 alpha-hydrolyase. Micromolar concentrations of insulin had similar effects to those of IGF-I. Moreover, the three factors that decreased testosterone production (EGF, bFGF and TGF beta 1) decreased gonadotropin receptor density, receptor mRNA levels and the mRNA levels for 17 alpha-hydroxylase. The potential effects of these growth factors on the transcription on the gonadotropin genes for the receptor and these three steroidogenic enzymes were measured by means of nuclear run-on assays. We demonstrated that the long-term inhibitory (EGF, bFGF, TGF beta 1) or stimulatory (IGF-I) effects of these growth factors are primarily due to a variation in the transcription rates of genes for the gonadotropin receptor, cytochrome P-450scc and 17 alpha-hydroxylase. Moreover, since previous studies have shown than some of these growth factors are expressed within the testis, they may play a physiological role in the regulation of differentiated testicular functions. PMID- 8706723 TI - Origin of the two carbonyl oxygens of bacteriochlorophyll a. Demonstration of two different pathways for the formation of ring E in Rhodobacter sphaeroides and Roseobacter denitrificans, and a common hydratase mechanism for 3-acetyl group formation. AB - A respiring culture of Rhodobacter sphaeroides, grown in the dark under defined aerobic conditions, produced cells capable of immediately commencing adaptation to photosynthetic growth on exposure to light and further reduction of oxygen tension. Adaptation was complete after 12 h and the bacteriochlorophyll a content increased 10-20-fold. This adaptation was performed in the presence of either H2(18)O or 18O2. The extracted bacteriochlorophyll a was examined by mass spectrometry to determine the origin of both the 3-acetyl adn 13(1)-oxo oxygen atoms: both were derived from water. The derivation of the 13(1)-oxo group from water in R. sphaeroides indicates that the formation of isocyclic ring E from the 13-propionic acid methylester side chain of Mg(2+)-protoporphyrin IX monomethylester is an anaerobic process involving a hydratase. This is very different to the situation in higher plants and green algae where the formation of isocyclic ring E is an aerobic process in which the 13(1)-oxo group is derived from molecular oxygen via an oxygenase. In contrast to adapting R. sphaeroides cells, the 13(1)-oxo group of bacteriochlorophyll a in growing cells of the obligate aerobic chemotrophic bacterium Roseobacter denitrificans, was labelled by 18O2 and is, therefore, derived from molecular oxygen like in higher plants and green algae; however, the 3-acetyl group was not labelled by 18O2. Thus, while the 13(1)-oxo group has different origins in R. sphaeroides and R. denitrificans, the 3-acetyl group arises in both bacteria by enzymic hydration of the vinyl group of a chlorophyll a derivative. PMID- 8706724 TI - Si-face stereospecificity at C5 of coenzyme F420 for F420-dependent glucose-6 phosphate dehydrogenase from Mycobacterium smegmatis and F420-dependent alcohol dehydrogenase from Methanoculleus thermophilicus. AB - Coenzyme F420 is a 5-deazaflavin. Upon reduction, 1,5-dihydro-coenzyme F420 is formed with a prochiral center at C5. In this study we report that the F420 dependent glucose-6-phosphate dehydrogenase from Mycobacterium smegmatis and the F420-dependent alcohol dehydrogenase from Methanoculleus thermophilicus are Si face stereospecific with respect to C5 of the 5-deazaflavin. These results were obtained by following the stereochemical course of the reversible incorporation of 3H into F420 from tritium-labeled substrates. Our findings bring to eight the number of coenzyme-F420-dependent enzymes shown to be Si-face stereospecific. No F420-dependent enzyme with Re-face stereospecificity is known. This is noteworthy since coenzyme F420 is functionally similar to pyridine nucleotides for which both Si-face and Re-face specific enzymes have been found. PMID- 8706725 TI - Analysis of phosphorylation and mutation of tyrosine residues of calmodulin on its activation of the erythrocyte Ca(2+)-transporting ATPase. AB - The role played by the phosphorylation sites of calmodulin on its ability to activate the human erythrocyte Ca(2+)-transporting ATPase (Ca(2+)-ATPase) was evaluated. Phosphorylation of mammalian calmodulin on serine/threonine residues by casein kinase II decreased its affinity for Ca(2+)-ATPase by twofold. In contrast, tyrosine phosphorylation of mammalian calmodulin by the insulin receptor kinase did not significantly alter calmodulin-stimulated Ca(2+)-ATPase activity. Two variant calmodulins, each containing only one tyrosine residue (the second Tyr is replaced by Phe) were also examined: [F138]calmodulin, a mutant containing tyrosine at position 99, and wheat germ calmodulin which has tyrosine at position 139. The concentrations of [F138]calmodulin and wheat germ calmodulin required for half-maximal activation of Ca(2+)-ATPase were tenfold and fourfold higher, respectively, than mammalian calmodulin. Phosphorylation at Tyr99 of [F138]calmodulin shifted its affinity for Ca(2+)-ATPase towards that of mammalian calmodulin. However, phosphorylation at Tyr139 of wheat germ calmodulin had essentially no effect on its interaction with Ca(2+)-ATPase. Thus, all of the observed effects of both phosphorylation and substitution of residues of calmodulin are on its affinity for Ca(2+)-ATPase, not on Vmax. The effects are dependent on the site of phosphate incorporation. Replacement of tyrosine with phenylalanine has a larger effect than phosphorylation of tyrosine, suggesting that the observed functional alterations reflect a secondary conformational change in the C-terminal half of calmodulin, the region that is important in its activation of Ca(2+)-ATPase. PMID- 8706726 TI - Regulation of membrane-type matrix metalloproteinase-1 expression by growth factors and phorbol 12-myristate 13-acetate. AB - Overexpression of membrane-type matrix metalloproteinase (MT-MMP-1) results in the activation of both endogenous and exogenous 72-kDa gelatinase. To understand the effects of MT-MMP-1 on 72-kDa gelatinase activation, we analyzed its expression in human fibroblasts and HT-1080 fibrosarcoma cells. Both cell types expressed the MT-MMP-1 mRNA constitutively at a considerable level and treatment of cells with PMA enhanced the expression about 2-3-fold. Concanavalin A treatment increased MT-MMP-1 mRNA levels in fibroblasts about 4-fold. Induction of MT-MMP-1 by phorbol 12-myristate 13-acetate (PMA) required protein synthesis as shown by cycloheximide inhibition. The induction was also inhibited by dexamethasone. Analysis of MT-MMP-1 mRNA stability using actinomycin D indicated that the half-life was rather long and not affected by PMA, suggesting transcriptional regulation. Only HT-1080 cells had significant 72-kDa gelatinase processing activity after treatment with PMA or concanavalin A, while fibroblasts were virtually negative. Immunoblotting analysis of fibroblast lysates indicated that MT-MMP-1 was present mainly in a 60-kDa form. PMA and concanavalin A caused 2-4-fold increases in its protein levels, while in HT-1080 cells PMA, concanavalin A, or overexpression of MT-MMP-1 did not significantly enhance the level of the 60-kDa protein. Instead, an immunoreactive, proteolytically processed 43-kDa form was observed, and its appearance correlated to 72-kDa gelatinase processing activity. Thus 72-kDa gelatinase activation, while enhanced by MT-MMP-1 expression, needs additional co-operating factors. PMID- 8706727 TI - Dipeptidyl-peptidase IV-beta, a novel form of cell-surface-expressed protein with dipeptidyl-peptidase IV activity. AB - The T-cell activation antigen CD26, is a type II membrane glycoprotein with intrinsic dipeptidyl-peptidase IV (DPP IV) activity, characterized by its capacity to cleave off N-terminal dipeptides containing proline as the penultimate residue. Independent of its catalytic activity, CD26 has also been characterized as adenosine deaminase binding protein. By using CD26 negative human C8166 cells, here we describe the existence of another cell-surface protein which manifests CD26-like DPP IV activity. For convenience, this protein will be referred to as DPP IV-beta. Consistent with the cell-surface expression of DPP IV beta, intact C8166 cells manifested a high level of DPP IV, whereas, they manifested poor activity against substrates of DPP II known to have an intracellular localization. A partially purified preparation of CD26 from human MOLT4 cells, and the DPP IV-beta expressed on intact cells were found to possess similar catalytic activity and pH optimum. In addition, cell-surface CD26 and DPP IV-beta on intact MOLT4 and C8166 cells, respectively, resisted digestion by proteolytic enzymes such as trypsin and proteinase K. However, adenosine deaminase activity was not detectable on the surface of C8166 cells in contrast to CD26 positive MOLT4 cells. In accord with this, 125I-labeled adenosine deaminase which binds CD26 was found not to bind DPP IV-beta. Gel-filtration experiments using 0.5% Triton X-100 extracts from C8166 and MOLT4 cells, revealed that the apparent molecular mass of DPP IV-beta is 82 kDa, whereas that of CD26 is 110 kDa as expected. Taken together, our results suggest that DPP IV-beta is a CD26-like protein which could be characterized by distinct properties. PMID- 8706728 TI - Frustulins: domain conservation in a protein family associated with diatom cell walls. AB - The outstanding feature of a diatom is the species-specific design and ornamentation of the silica-based cell wall, termed frustulum. A new frustulum is shaped in a specialized organelle (silica deposition vesicle) and secreted. Proteins in the lumen of this organelle may control the biomineralization process and are likely to remain associated with the mature cell wall. Therefore a study of the structures of proteins associated with the diatom cell wall was initiated. The complete primary structures of three cell wall proteins (denoted as frustulins) have been determined. In addition, partial amino acid sequences from two more cell wall components were obtained. From these data, a highly conserved domain has been identified as a common building block of diatom cell wall proteins that is repeated several times per polypeptide chain together with polyproline/hydroxyproline or polyglycine spacers. All frustulins characterized so far, are synthesized as preproteins with a novel type of N-terminal presequence. PMID- 8706729 TI - Limited proteolysis and amino acid replacements in the effector region of Thermus thermophilus elongation factor Tu. AB - The effector region of the elongation factor Tu (EF-Tu) from Thermus thermophilus was modified by limited proteolysis or via site-directed mutagenesis. The biochemical properties of the obtained EF-Tu variants were investigated with respect to partial reactions of the functional cycle of EF-Tu. EF-Tu that was cleaved at the Arg59-Gly60 peptide bond [EF-Tu-(1-59)/EF-Tu-(60-405)] bound GDP, EF-Ts and aminoacyl-tRNA, had normal intrinsic GTPase activity and was active in poly(U)-dependent poly(Phe) synthesis. However, the GTPase activity of EF-Tu-(1 59)/EF-Tu-(60-405) was not stimulated by T. thermophilus 70S ribosomes, and its GTP-dissociation rate was increased compared with that of intact EF-Tu. EF-Tu cleaved at the Lys52-Ala53 peptide bond has properties similar to EF-Tu-(1-59)/EF Tu-(60-405). By means of site-directed mutagenesis, Glu55 was replaced by Leu, Glu56 by Ala and Arg59 by Thr in T. thermophilus EF-Tu. These amino acid substitutions did not substantially affect either the affinity of EF-Tu. GTP for aminoacyl-tRNA or the interactions with GDP, GTP or EF-Ts. Similarly the intrinsic GTPase activity is not influenced. Replacement of Glu56 by Ala led to strong reduction in the ribosome-induced GTPase activity. This effect is specific since replacement of the neighbouring Glu55 by Leu did not affect the ribosome induced GTPase activity. The results demonstrate that the structure of the effector region of EF-Tu in the vicinity of Arg59 is important for the control of the GTPase activity by ribosomes. PMID- 8706730 TI - Clavin, a type-1 ribosome-inactivating protein from Aspergillus clavatus IFO 8605. cDNA isolation, heterologous expression, biochemical and biological characterization of the recombinant protein. AB - We describe the cloning and expression of a new cDNA from the filamentous fungus Aspergillus clavatus IFO 8605. This cDNA contains an open reading frame (ORF) that predicts a putative ribonuclease precursor with high similarity to the alpha sarcin family of ribosome-inactivating proteins (RIPs). The cDNA encoding the mature protein was expressed in Escherichia coli, and the recombinant protein, a 17-kDa polypeptide designated clavin was purified and characterized. Clavin shows typical type-1 RIP properties: specific cleavage of ribosomal and synthetic RNA and inhibition of protein synthesis in cell-free and cellular systems. When selectively targeted to a tumour cell antigen by coupling to a monoclonal antibody (mAb) clavin was able to inhibit protein synthesis at nanomolar concentration. Pharmacokinetics analysis in mice indicated an elimination half life (t1/2 beta) of 7.4 h with no particular accumulation in major organs. Liver toxicity was very limited and transient while no alteration of kidney function was observed. Clavin induced a late and very low antibody response in mice. The in vitro and in vivo biological characteristics of clavin, together with its availability in large amounts, suggest the usefulness of this toxin in the production of toxic chemical conjugates. PMID- 8706731 TI - Bean ribonuclease-like pathogenesis-related protein genes (Ypr10) display complex patterns of developmental, dark-induced and exogenous-stimulus-dependent expression. AB - The intracellular pathogenesis-related (PR) proteins of common bean (Phaseolus vulgaris L.) are encoded by a highly polymorphic family of at least 20 genes. One member, the Ypr10*c gene, has been isolated and characterised. The deduced amino acid sequence of the encoded protein, PR-10, exhibits similarities to tree-pollen allergens, to food allergens from celery and apple and to ginseng ribonuclease peptide sequences. We show by RNA blot analysis that the Ypr10 gene family, including Ypr10*c, is strongly expressed in bean roots. In leaves Ypr10 transcript levels are low in young and mature stages but are elevated during senescence and in diseased states. Dark treatment of leaves causes strong induction of Ypr10 transcripts, which is reversible by light, and diurnal rhythms of transcript accumulation during the night are observed. Ypr10 genes are responsive to external stimuli related to pathogen-defence such as glutathione or salicylic acid. Transcriptional activity of a Ypr10*c promoter-beta-glucuronidase fusion gene in transgenic tobacco was observed in roots, in developing xylem and phloem of stems, and in the blade of senescent leaves, with highest levels at the onset of senescence. The most striking characteristic of developmental expression was the specific localisation of beta-glucuronidase activity in the transmitting tract of styles in flowers at anthesis. Feeding of various pathogen-related and stress-related stimuli to young tobacco leaves led to accumulation of GUS activity in leaf blades. We identify considerable spatio-temporal similarities between reported expression patterns of Ypr10 genes and ribonuclease genes, which, together with the significant sequence similarity to the ginseng ribonuclease, support the hypothesis of a ribonuclease function for PR-10 proteins and allow the prediction of possible biological roles. PMID- 8706732 TI - Temperature-dependent conformational change of bacteriorhodopsin as studied by solid-state 13C NMR. AB - Cross-polarization and dipolar-decoupled magic-angle spinning 13C-NMR spectra of [3-13C]Ala-labelled bacteriorhodopsin were obtained for hydrated purple membrane in the temperatures range 23 degrees C to -110 degrees C. Well-resolved 13C-NMR signals were observed either at ambient temperature or at -20 degrees C but were broadened considerably at lower temperature below -40 degrees C. This situation was interpreted in terms of the presence of exchange processes with a rate constant of 10(2) s-1 at ambient temperature among several conformations slightly different from each other. We found that such an exchange process was strongly influenced by the manner of organization of the lipid bilayers depending upon the presence or absence of cations responsible for electric shielding of negative charge at the polar head groups. The manner of organization of the lipid bilayers was conveniently characterized by a characteristic temperature at which the methyl peaks of fatty acyl groups of lipids in the purple membrane were suppressed due to interference of motional frequency with the decoupling frequency (10-100 kHz) for preparations containing 10 mM NaCl or CaCl2. No such spectral change in the absence of these cations was noted even if a preparation was cooled to -110 degrees C. The secondary structures of [3-13C]Ala-labelled bacteriorhodopsin was not always identical at temperatures between ambient and low temperatures, since the 13C chemical shifts and relative peak intensities for purple membrane preparations containing these salts changed with temperature in the range -110 degrees C to 23 degrees C. In particular, we found that some residues involving Ala residues at the alpha II-helix and loop region were converted at temperatures below -60 degrees C to a conformation involving alpha 1 helix. In other words, some portion of the alpha-helical conformation of bacteriorhodopsin proposed from results obtained by cryo-electron microscopy, at very low temperatures, is not always retained at ambient temperature. PMID- 8706733 TI - Rat pristanoyl-CoA oxidase. cDNA cloning and recognition of its C-terminal (SQL) by the peroxisomal-targeting signal 1 receptor. AB - The composite pristanoyl-CoA oxidase cDNA sequence, derived from two overlapping clones from a rat liver cDNA library and a 5'-RACE (rapid amplification of cDNA ends) PCR fragment, consisted of 2600 bases and contained an open reading frame of 2100 bases, encoding a protein of 700 amino acids with a calculated molecular mass of 78445 Da. This value is somewhat larger than the reported molecular mass of 70 kDa as determined earlier by SDS-gel electrophoresis. The amino acid identity with rat palmitoyl-CoA oxidase was rather low (28%) and barely higher than that with the yeast acyl-CoA oxidases (20%), suggesting that the palmitoyl CoA oxidase/pristanoyl-CoA oxidase duplication occurred early in evolution. The carboxy-terminal tripeptide of pristanoyl-CoA oxidase was SQL. In vitro studies with the bacterially expressed human peroxisomal-targeting signal-1 import receptor indicated that SQL functions as a peroxisome-targeting signal. Northern analysis of tissues from control and clofibrate treated rats demonstrated that the pristanoyl-CoA oxidase gene is transcribed in liver and extrahepatic tissues and that transcription is not enhanced by treatment of rats with peroxisome proliferators. No mRNA could be detected by northern analysis of human tissues, suggesting that the human pristanoyl-CoA oxidase gene, if present, is only poorly or not transcribed. PMID- 8706734 TI - Identification and characterization of a rice cysteine endopeptidase that digests glutelin. AB - Little or no endopeptidase activity was detected in extracts from storage organs of dark-grown rice seeds until day 6 of post-imbibition, and the activity expressed per seed increased notably after day 9, reached a maximum on day 18, then decreased. Two major endopeptidases, REP-1 and REP-2, were present in the 40 75% saturated ammonium sulfate fraction from day-9 germinated seeds, and could be separated by hydrophobic column chromatography. REP-1 was further purified to a single polypeptide of 36 kDa. REP-1 digested in vitro both the acidic and basic subunits of rice glutelin, the major seed storage protein of rice. Determination of the N-terminal amino acid sequence and experiments with protease inhibitors indicated that REP-1 is a cysteine endopeptidase. The nucleotide sequence of a full-length REP-1 cDNA was determined by a combination of screening of cDNA libraries from rice seeds and the 5' rapid amplification of cDNA ends technique. PMID- 8706735 TI - Identification of protein-phosphatase-1-binding domains on the glycogen and myofibrillar targetting subunits. AB - The specificity of the catalytic subunit of protein phosphatase-1 (PP1c) is modified by regulatory subunits that target it to particular subcellular locations. Here, we identify PP1c-binding domains on GL and GM, the subunits that target PP1c to hepatic and muscle glycogen, respectively, and on M110, the subunit that targets PP1c to smooth muscle myosin. GM-(G63-T93) interacted with PP1c and prevented GL from suppressing the dephosphorylation of glycogen phosphorylase, but it did not dissociate GL from PP1c or affect other characteristic properties of the PP1GL complex. These results indicate that GL contains two PP1c-binding sites, the region which suppresses the dephosphorylation of glycogen phosphorylase being distinct from that which enhances the dephosphorylation of glycogen synthase. At higher concentrations, GM (G63-N75) had the same effect as GM-(G63-T93), but not if Ser67 was phosphorylated by cyclic-AMP-dependent protein kinase. Thus, phosphorylation of Ser67 dissociates GM from PP1c because phosphate is inserted into the PP1c binding domain of GM. M110-(M1-E309) and M110-(M1-F38), but not M110-(D39-E309), mimicked the M110 subunit in stimulating dephosphorylation of the smooth muscle myosin P-light chain and heavy meromyosin in vitro. However, in contrast to the M110 subunit and M110-(M1-E309), neither M110-(M1-F38) nor M110-(D39-E309) suppressed the PP1c-catalysed dephosphorylation of glycogen phosphorylase. These observations suggest that the region which stimulates the dephosphorylation of myosin is situated within the N-terminal 38 residues of the M110 subunit, while the region which suppresses the dephosphorylation of glycogen phosphorylase requires the presence of at least part of the region 39-309 which contains seven ankyrin repeats. M110-(M1-F38) displaced GL from PP1c, while GM-(G63-T93) displaced M110 from PP1c in vitro. These observations indicate that the region(s) of PP1c that interact with GM/GL and M110 overlap, explaining why different forms of PP1c contain just a single targetting subunit. PMID- 8706736 TI - Regions of the 110-kDa regulatory subunit M110 required for regulation of myosin light-chain-phosphatase activity in smooth muscle. AB - To characterize the in situ interactions between the subunits (regulatory 110 kDa, M110; 21-kDa, M21 and catalytic, 37-kDa, PP1c) of smooth muscle myosin phosphatase (SMPP-1M), we determined, in Triton-X-100-permeabilized rabbit portal vein contracted with microcystin-LR, the ability of the following fragments of M110 to regulate relaxation induced by exogenous PP1c: (a) M110 purified from pig bladder; (b) the 72.5-kDa N-terminal fragment expressed from rat kidney cDNA [glutathione-S-transferase-M110-(11-612)-peptide]; (c) a 58-kDa fragment, the N terminal degradation product of M110 (M58); (d) two fragments expressed from rat aorta cDNA [M110-(1-309)-peptide and M110-(39-309)-peptide]; a synthetic fragment of M110 [M110-(1-38)-peptide]. The M110/M21 complex accelerated approximately 1.6 fold the rate of dephosphorylation of the myosin P-light chain and also relaxation induced by PP1c. The glutathione-S-transferase-M110-(11-612)-peptide and the M58 fragments, as well as the M110-(1-309)-peptide and, at higher concentration, M110-(1-38)-peptide, had similar effects that did not require the M21 subunit. Arachidonic acid, known to dissociate PP1c from the native holoenzyme and inhibit SMPP-1M activity, inhibited the regulatory action of the M110/M21 complex on PP1c activity and, to a lesser extent that of the glutathione S-transferase-M110-(11-612)-peptide, but not that of the M58 fragment or of the shorter peptides. We conclude that, consistent with in vitro studies [8], the N terminal sequence (1-309) of the M110 subunit is also sufficient to enhance the activity of PP1c for myosin in muscle. However, its C-terminal half (downstream from the M58 fragment) is required for inhibition by arachidonic acid. In contrast to the effect of the M110 subunit and its fragments, a peptide, corresponding to part of the PP1c-binding site of the regulatory glycogen-binding subunit from skeletal muscle GM [GM-(63-93)-peptide], specifically slowed the relaxation, induced by flash photolysis of diazo-2, of Triton X-100-permeabilized femoral artery strips, and inhibited the holoenzyme-induced relaxation in the portal vein, suggesting that the GM subunit can compete with the regulatory effect of M110 on PP1c in smooth muscle. PMID- 8706737 TI - Domain structure, stability and interactions in streptokinase. AB - The structural organization of streptokinase was established through detailed study of its denaturation by differential scanning calorimetry. Streptokinase exhibited a complex endotherm whose shape was sensitive to changing pH. In all cases the endotherms were easily described by four two-state transitions indicating unambiguously the presence of four independently folded domains in the molecule. Two of them were slightly destabilized by lowering pH from 7.0 to 3.8 while the other two were stabilized in this pH range. Two proteolytic fragments of streptokinase were examined, a 37-kDa fragment beginning at Ile1 with a cleavage following Phe62, and a 17-kDa fragment beginning at Lys 147. At pH 8.5, three two-state transitions were observed in the former and two in the latter indicating this many domains in each and suggesting that the fragments are formed by a step-wise removal of individual domains from the parent molecule. Comparison of the melting of these fragments with that of streptokinase allowed the first two transitions in the parent protein to be assigned to the melting of two NH2 terminal domains and the two higher-temperature transitions to the melting of the two COOH-terminal domains. The latter two domains strongly interact with each other since the absence of the most stable extreme COOH-terminal domain in both fragments resulted in a strong destabilization of its neighbor whose melting occurred with a midpoint near room temperature. The two NH2-terminal domains seem to be more independent. One of them melts similarly in the parent protein and both fragments while the other, formed by the 1-146 region, is less stable in the 37-kDa fragment. This destabilization is most probably due to the cleavage after Phe62 which, based on the sequence similarity of streptokinase with serine proteases, may be part of a surface-oriented loop. PMID- 8706738 TI - Recombinant soluble beta-1,4-galactosyltransferases expressed in Saccharomyces cerevisiae. Purification, characterization and comparison with human enzyme. AB - beta-1,4-Galactosyltransferase (Gal-T, EC 2.4.1.38) transfers galactose (Gal) from UDP-Gal to N-acetyl-D-glucosamine or a derivative GlcNAc-R. Soluble Gal-T, purified from human breast milk, was shown to be very heterogeneous by isoelectric focusing (IEF). In order to produce sufficient homogeneous enzyme for three-dimensional analysis, the human enzyme (hGal-T) has been expressed in Saccharomyces cerevisiae, production scaled up to 187 U recombinant Gal-T (rGal T) and purified. The purification protocol was based on chromatography on concanavalin-A-Sepharose followed by affinity chromatographies on GlcNAc Sepharose and alpha-lactalbumin-Sepharose. Analysis by SDS/PAGE revealed hyperglycosylation at the single N-glycosylation site, preventing recognition by antibodies. Analysis by IEF revealed considerable heterogeneity of rGal-T. The N glycan could be removed by treatment with endoglycosidase H (endo H). The N deglycosylated form of rGal-T retained full activity and showed only three isoforms by IEF analysis. Then we abolished the single N-glycosylation consensus sequence by site-directed mutagenesis changing Asn69-->Asp. The soluble mutated enzyme (N-deglycosylated rGal-T) was expressed in S. cerevisiae and its production scaled up to 60 U.N-deglycosylated rGal-T was purified to electrophoretic homogeneity. When analyzed by IEF, N-deglycosylated rGal-T was resolved in two bands. The O-glycans could be removed by jack bean alpha mannosidase treatment and the completely deglycosylated Gal-T appeared homogeneous by IEF. The kinetic parameters of N-deglycosylated rGal-T were shown not to differ to any significant extent from those of the hGal-T. No significant changes in CD spectra were observed between hGal-T and N-deglycosylated rGal-T. Light-scattering analysis revealed dimerization of both enzymes. These data indicate that N-deglycosylated rGal-T was correctly folded, homogeneous and thus suitable for crystallization experiments. PMID- 8706739 TI - Interaction of amino acid residues at positions 8-15 of secretin with the N terminal domain of the secretin receptor. AB - The ability of secretin, PACAP-(1-27)-peptide, and ten hybrid peptides to recognize and activate the rat secretin and vasoactive intestinal polypeptide (PACAP type II VIP1) receptors was tested on recombinant Chinese hamster ovary (CHO) cell lines. PACAP had a 2500-fold lower affinity than secretin for the secretin receptor, and secretin had a 300-fold lower affinity than PACAP for the VIP1 receptor. Amino acids 8, 13, and 15 of the PACAP molecule contributed significantly to the low affinity of PACAP for the secretin receptor. The amino acids at positions 5, 9, 10, 15, 16, and unidentified amino acid(s) between positions 17-20 made limited contributions to the low affinity of secretin for the VIP1 receptor. To identify the receptor region that interacts with these amino acids, we constructed chimeric receptors, which consist either of the N terminal extracellular part of the secretin receptor and the core of the VIP1 receptor (N-Sn/VIP1r) or the N-terminal extracellular part of the VIP1 receptor and the core of the secretin receptor (N-VIP1/Snr), and tested the ability of the hybrid ligands to activate the adenylate cyclase of CHO cells expressing these chimeric receptors. The N-Sn/VIP1 receptors had a higher affinity for secretin than for PACAP. The hybrid peptide 6 that consists of the PACAP-(1-8)-Sn-(9-15) PACAP-(16-27)-peptide sequence had a 30-fold to 200-fold higher potency than either parent peptide for the chimeric receptor, which suggests that while the N- and/or C-terminal part of the peptide interact with the transmembrane domain of the receptor, the discriminator region 9-15 recognizes the extracellular N terminal domain of the receptor. This was confirmed by the observation that, out of all the peptides tested, hybrid 6 had the weakest potency for activation of the N-VIP1/Sn chimeric receptors. PMID- 8706740 TI - The hemolysin B protein, expressed in Saccharomyces cerevisiae, accumulates in binding-protein (BiP)-containing structures. AB - The hemolysin B (Hly B) protein of Escherichia coli, a member of the ABC transporter family, was expressed in Saccharomyces cerevisiae and tested for its ability to complement a defect in the a-factor transporter Ste6. We found that HlyB was not able to restore mating ability to a STE6 deletion strain. The HlyB protein did not co-fractionate with Ste6 on sucrose gradients, indicating that improper localization of the HlyB protein could contribute to the lack of complementation. Immunofluorescence experiments suggest that HlyB is localized to structures derived from the endoplasmic reticulum (ER). The HlyB-expressing cells revealed a perinuclear staining typical of ER-localized proteins and intensely staining ring-like structures (HlyB-bodies). Double-label immunofluorescence experiments show that the HlyB structures also contain the ER binding protein (BiP), the product of the KAR2 gene. The HlyB protein, however, did not co fractionate with another ER marker protein, the NADPH cytochrome c reductase. The HlyB bodies could be derivatives of a novel compartment of the early secretory pathway which contains BiP but not other resident ER proteins. In this case, HlyB could serve as a tool for the biochemical characterization of this compartment. PMID- 8706741 TI - Characterization of the interaction of the monomeric GTP-binding protein Rab3a with geranylgeranyl transferase II. AB - The monomeric GTP-binding protein Rab3a controls exocytosis in neuroendocrine and neuronal cells. Like other members of the Rab family, Rab3a is posttranslationally modified by the addition of hydrophobic geranylgeranyl groups to its C-terminus. The geranylgeranylation reaction is catalysed by the heterotrimeric geranylgeranyl transferase II. We describe the cDNA cloning of the beta-subunit of human geranylgeranyl transferase II by means of the yeast two hybrid system. The human enzyme, which is 49% and 96% similar to yeast and rat isoforms, respectively, can complement the beta-subunit deficiency in the yeast strain ANY119. Furthermore, by means of the two-hybrid system and in vitro geranylgeranylation reactions with purified recombinant rat geranylgeranyl transferase II, we have characterized Rab3a domains implicated in the interaction with geranylgeranyl transferase II. We find that the N-terminus, the effector loop, the hypervariable region of the C-terminus, and the geranylgeranyl-acceptor cysteines have roles in this interaction. The GDP-bound form of Rab3a is the preferred substrate of geranylgeranyl transferase II. PMID- 8706742 TI - In vitro activation of the NADPH oxidase by fluoride. Possible involvement of a factor activating GTP hydrolysis on Rac (Rac-GAP). AB - The possible mechanism of activation of the NADPH oxidase by fluoride was investigated in the cell-free system. It is shown that the stimulatory effect of fluoride is inhibited by guanosine 5'-O-(2-thiodiphosphate) (GDP[S]) and potentiated by GTP. The effect of fluoride is not additive with GTP[S]. Fluoride activation requires the presence of Mg2+ in millimolar concentration but is independent of Al3+. The activating effect of fluoride is preserved in solubilized membrane extract after removal of the majority of heterotrimeric GTP binding proteins by immunoadsorption. Fluoride has no direct action either on the nucleotide exchange of GTP hydrolysis of the isolated Rac protein. In contrast, fluoride effectively inhibits Rac-GTPase activity enhanced by a membrane component. In this way, fluoride could prolong the prevalence of Rac in the GTP bound state and, as a consequence, activate NADPH oxidase. The possibility of the involvement of a membrane-bound Rac GTPase-activating protein activity in the physiological regulation of the enzyme is raised. PMID- 8706743 TI - Intrinsic conformational preferences of the Hoechst dye family and their influence of DNA binding. AB - Quantum mechanical calculations have been used to investigate the molecular conformation of the Hoechst family of DNA-binding dyestuffs. Compounds in which the phenolic substituent adopts either a meta or para position were studied. Two different environments have been considered, which are the gas phase and aqueous solution; the conformation in aqueous solution has been modeled through a self consistent reaction field strategy. The results clearly indicate that Hoechst dyes do not adopt a planar conformation and that the degree of planarity is controlled by the external environment. A comparison with experimental data reveals that the conformation of Hoechst dyes in the gas phase is similar to that observed in DNA complexes by X-ray crystallography. In aqueous solution, the conformation deviates from planarity more than in the gas phase, since non-bonded interactions with the solvent offset the loss of conjugative interactions. The role of the drug conformation in the binding mechanism with DNA is discussed. PMID- 8706744 TI - A 30-kDa host protein binds to two very-late baculovirus promoters. AB - A 30-kDa host factor (polyhedrin-promoter-binding protein; PPBP) specifically binds to sequences critical for transcription from the baculovirus polyhedrin (p29) gene initiator promoter [Burma, S., Mukherjee, B., Jain, A., Habib, S. & Hasnain, S. E. (1994) J. Biol. Chem. 269, 2750-2757; Mukherjee, B., Burma, S. & Hasnain, S. E. (1995) J. Biol. Chem. 270, 4405-4411]. A host factor also binds, in gel shift assays, to the very-late p10 gene promoter through DNA sequence motifs similar to the PPBP.p29 interaction. The p10 host factor complex was specifically competed out with oligonucleotides containing p29 cognate sequence motifs AATAAA and TAAGTATT, but this did not occur when these motifs were replaced with random sequences. From ultraviolet cross-linking analysis, the molecular mass of this host factor was estimated to be approximately 30 kDa. Experiments were performed to investigate if this host factor displayed any differences in affinity and turnover with respect to the p29 and p10 untranslated leader sequences known to be important for temporal fine tuning and the late burst of transcription. Half-life determination of the p10-binding protein revealed similar binding affinities for the initiator elements of both the promoters, but higher affinity for the p10 5'-untranslated region (approximately 30 min versus approximately 10 min). The involvement of a similar host factor binding to both the p10 and p29 promoters indicates the possibility of a similar mode of transcription initiation from these two very-late promoters. PMID- 8706745 TI - Dimethylsulfide:acceptor oxidoreductase from Rhodobacter sulfidophilus. The purified enzyme contains b-type haem and a pterin molybdenum cofactor. AB - Dimethylsulfide:receptor oxidoreductase was purified from the purple non-sulfur phototrophic bacterium Rhodobacter sulfidophilus. The native form of the enzyme had a molecular mass of 152 kDa and was composed of three distinct subunits of 94, 38 and 32 kDa. Dimethylsulfide:acceptor oxidoreductase did not oxidise other thioethers which were tested. The enzyme was able to reduce a variety of N-oxides using reduced methylviologen as electron donor but it reduced dimethylsulfoxide at a very low rate. The resting form of dimethylsulfide:acceptor oxidoreductase exhibited a spectrum which was characteristic of a reduced cytochrome with absorbance maxima at 562 nm, 533 nm and 428 nm. Pyridine haemochrome analysis established that the cytochrome contained a b-type haem and a content of 0.65 mol protohaem/mol enzyme was determined. After oxidation of the haem with ferricyanide, the absorbance spectrum of the reduced cytochrome was restored by reduction with dimethylsulfide. Metal analysis revealed that dimethylsulfide:acceptor oxidoreductase contained 0.5 mol Mo and 3.5 mol Fe/mol enzyme. Heat treatment of the enzyme released material with fluorescence excitation and emission spectra which were characteristic of form B of the pterin component of the pterin molybdenum cofactor. From this analysis it is concluded that dimethylsulfide:acceptor oxidoreductase is a molybdenum oxotransferase which may also contain a iron-sulfur cluster. It is suggested that the haem and pterin molybdenum cofactor are associated with the 94-kDa subunit. PMID- 8706746 TI - An essential lysine in the substrate-binding site of ornithine carbamoyltransferase. AB - Treatment of ornithine carbamoyltransferase from dolphin Stenella with pyridoxal phosphate, followed by reduction with NaBH4 resulted in complete loss of enzyme activity. The phosphate alone or the substrate analogue 2-aminovaleric acid moderately decreased the extent of inactivation, while carbamoyl phosphate plus 2 aminovaleric acid provided complete protection from inactivation. The partially inactivated enzyme showed K(m) values for substrates equivalent to those of native enzyme and lowered Kcat values. Two lysyl residues were substantially modified in the absence of ligands but only one of them was responsible for the inactivation of catalytic activity. Modification of a single subunit was sufficient to completely abolish the catalytic activity of the trimeric enzyme. The lysine involved has been identified as lysine 56 on the known primary structure of homologous human liver enzyme. PMID- 8706747 TI - Probing electron transfer in flavocytochrome P-450 BM3 and its component domains. AB - Rapid events in the processes of electron transfer and substrate binding to cytochrome P-450 BM3 from Bacillus megaterium and its constituent haem-containing and flavin-containing domains have been investigated using stopped-flow spectrophotometry. The formation of a blue semiquinone flavin form occurs during the NADPH-dependent reduction of the flavin domain and a species with a similar absorption maximum is also seen during reduction of the holoenzyme by NADPH. EPR spectroscopy confirms the formation of the flavin semiquinone. The formation of this semiquinone is transient during fatty acid monooxygenation by the holoenzyme, but in the presence of excess NADPH the species reforms once fatty acid is exhausted. Electron transfers through the reductase domain are too rapid to limit the fatty acid monooxygenation reaction. The substrate-binding-induced haem iron spin-state shift also occurs much faster than the Kcat at 25 degrees C. The rate of first electron transfer to the haem domain is also rapid; but it is of the order of 5-10-times larger than the Kcat for the enzyme (dependent on the fatty acid used). Given that two successive electron transfers to haem iron are required for the oxygenation reaction, these rates are likely to exert some control over the rate of fatty acid oxygenation reactions. The presence of large amounts of NADPH also results in decreased rates of electron transfer from flavin to haem iron. In the difference spectrum of the active fatty acid hydroxylase, features indicative of a high-spin iron haem accumulate. These are in accordance with the presence of large amounts of an Fe(3+)-product bound enzyme during turnover and indicate that product release may also contribute to rate limitation. Taken together, these data suggest that the catalytic rate is not determined by the accumulation of a single intermediate in the reaction scheme, but rather that it is controlled in a series of steps. PMID- 8706748 TI - Protein engineering studies of dichloromethane dehalogenase/glutathione S transferase from Methylophilus sp. strain DM11. Ser12 but not Tyr6 is required for enzyme activity. AB - The structural gene for dichloromethane dehalogenase/glutathione S-transferase (GST, EC 2.5.1.18) from Methylophilus sp. strain DM11 was subcloned into a multicopy plasmid under the control of the T7 polymerase promoter, allowing expression in Escherichia coli and easy purification of the enzyme in good yield. Several point mutations leading to amino acid changes at residues Tyr6, His8 and Ser12 of the protein were introduced in this gene. Mutations at Tyr6, the N terminal tyrosine known to be essential for enzymatic activity in glutathione S transferases of the alpha, mu, and pi classes, had little effect on the activity of dichloromethane dehalogenase. The same applied for mutations at residue His8, which from multiple alignments of GST sequences may also correspond to the conserved N-terminal tyrosine residue of GST enzymes. The higher turnover rate of the wild-type enzyme with dibromomethane compared with dichloromethane was lost in mutants with amino acid replacements at residue His8, but retained in mutant proteins at Tyr6. Mutations at Ser12 led to mutants with drastically reduced enzymatic activity, pinpointing this residue as an essential determinant of catalytic efficiency. PMID- 8706750 TI - L-aspartate oxidase from Escherichia coli. II. Interaction with C4 dicarboxylic acids and identification of a novel L-aspartate: fumarate oxidoreductase activity. AB - L-Aspartate oxidase is a monomeric flavoprotein that catalyzes the first step in the de novo biosynthetic pathway for pyridine nucleotide formation under both aerobic and anaerobic conditions. In spite of the physiological importance of this biosynthesis in particular in facultative aerobic organisms, such as Escherichia coli, little is known about the electron acceptor of reduced L aspartate oxidase in the absence of oxygen. In this report, evidence is presented which suggests that in vitro fumarate can play such a role. L-Aspartate oxidase binds succinate and fumarate with Kd values of 0.24 mM and 0.22 mM, respectively. A competitive behaviour was observed for these two dicarboxylic acids towards iminoaspartate and sulfite ions. Photoreduction experiments suggest that fumarate and succinate bind at or close to the active site of the molecule. A new fumarate reductase activity of L-aspartate oxidase is reported using benzylviologen or L aspartate as reductants and fumarate as oxidant. Steady-state kinetics for the oxidase and the fumarate reductase activity of L-aspartate oxidase were obtained using either fumarate or oxygen as electron acceptor and L-aspartate as electron donor. Finally, succinate was identified as the product of the L aspartate:fumarate oxidoreductase activity using radiolabeled fumarate under anaerobic conditions. The results suggest that fumarate can be a valuable alternative to oxygen as a substrate for L-aspartate oxidase. PMID- 8706749 TI - L-aspartate oxidase from Escherichia coli. I. Characterization of coenzyme binding and product inhibition. AB - This paper reports the biochemical characterization of the flavoprotein L aspartate oxidase from Escherichia coli. Modification of a previously published procedure allowed overexpression of the holoenzyme in an unproteolysed form. L Aspartate oxidase is a monomer of 60 kDa containing 1 mol of noncovalently bound FAD/mol protein. A polarographic and two spectrophotometric coupled assays have been set up to monitor the enzymatic activity continuously. L-Aspartate oxidase was subjected to product inhibition since iminoaspartate, which results from the oxidation of L-aspartate, binds to the enzyme with a dissociation constant (Kd) equal to 1.4 microM. The enzyme binds FAD by a simple second-order process with Kd 0.67 microM. Site-directed mutagenesis of the residues E43, G44, S45, F47 and Y48 located in the putative binding site of the isoallossazinic portion of FAD reduces the affinity for the coenzyme. PMID- 8706751 TI - The O-specific polysaccharide chain of Campylobacter fetus serotype B lipopolysaccharide is a D-rhamnan terminated with 3-O-methyl-D-rhamnose (D acofriose). AB - An O-specific polysaccharide was liberated from Campylobacter fetus subsp. fetus serotype B lipopolysaccharide by mild acid hydrolysis followed by gel chromatography. This polysaccharide was found to contain D-rhamnose and 3-O methyl-D-rhamnose (D-Rha3Me, D-acofriose) in a ratio of approximately 24:1, as well as lipopolysaccharide core constituents. The structure of the polysaccharide was studied by 1H-NMR and 13C-NMR spectroscopy, which included two-dimensional COSY, rotating-frame NOE spectroscopy (ROESY), and computer-assisted analysis of the 13C-NMR spectrum. Methylation analysis using [2H3]methyl iodide and Smith degradation followed by GLC/MS of the derived acetylated oligosaccharide-alditols was used to determine the location of D-acofriose. The O-specific polysaccharide is linear, consists on average of 12 disaccharide repeating units, and is terminated by a residue of D-acofriose. The following structure of the D-rhamnan chain was established: [sequence: see text] PMID- 8706752 TI - In vivo phosphorylation of the purine/cytosine permease from the plasma membrane of the yeast Saccharomyces cerevisiae. AB - The purine/cytosine permease, encoded by the FCY2 gene, is a carrier located in the plasma membrane of the yeast Saccharomyces cerevisiae. Polyclonal antibodies were raised against two peptides that corresponded to the sub-N-terminal and C terminal sequences of the putative protein deduced from the FCY2 gene. Immunoprecipitation experiments performed with protein extracts labelled in vivo with 35S showed that purine/cytosine permease is specifically detected as a broad and diffuse band. The apparent molecular mass of this protein was 45-50 kDa. By means of in vivo pulse/chase 35S-labelling experiments, we observed a slight increase in the apparent molecular mass of purine/cytosine permease during the chase. This shift in electrophoretic mobility of the protein suggested a post translational modification. This molecular mass increase was eliminated by alkaline phosphatase treatment of the immunoprecipitate, which strongly suggested phosphorylation of the carrier. This proposal was confirmed by in vivo [32P]P(i) labelling and immunoprecipitation of purine/cytosine permease with purified anti (sub-N-terminal peptide) IgG or anti-(C-terminal peptide) IgG. Phosphoamino acid analysis indicated that phosphorylation occurred on seryl residues of purine/cytosine permease. By means of thermosensitive secretory-pathway-mutant strains, we demonstrated that purine/cytosine permease phosphorylation occurred either between the Golgi apparatus and the plasma membrane or in the plasma membrane itself. PMID- 8706753 TI - Activation of the uncoupling protein by fatty acids is modulated by mutations in the C-terminal region of the protein. AB - The transport properties of the uncoupling protein (UCP) from brown adipose tissue have been studied in mutants where Cys304 has been replaced by either Gly, Ala, Ser, Thr, Ile or Trp. This position is only two residues away from the C terminus of the protein, a region that faces the cytosolic side of the mitochondrial inner membrane. Mutant proteins have been expressed in Saccharomyces cerevisiae and their activity determined in situ by comparing yeast growth rates in the presence and absence of 2-bromopalmitate. Their bioenergetic properties have been studied in isolated mitochondria by determining the effects of fatty acids and nucleotides on the proton permeability and NADH oxidation rate. It is revealed that substitution of Cys304 by non-charged residues alters the response of UCP to fatty acids. The most effective substitution is Cys for Gly since it greatly enhances the sensitivity to palmitate, decreasing threefold the concentration required for half-maximal stimulation of respiration. The opposite extreme is the substitution by Ala which increases twofold the half maximal concentration. We conclude that the C-terminal region participates in the fatty acid regulation of UCP activity. The observed correlation between yeast growth rates in the presence of bromopalmitate and the calculated activation constants for respiration in isolated mitochondria validates growth analysis as a method to screen the in situ activity of UCP mutants. PMID- 8706754 TI - Transcriptional induction of rat liver apolipoprotein A-I gene expression by glucocorticoids requires the glucocorticoid receptor and a labile cell-specific protein. AB - Treatment with glucocorticoids increases the concentration of plasma high-density lipoprotein (HDL), which is inversely correlated to the development of atherosclerosis. Previously, we demonstrated that repeated administration of glucocorticoids increases apolipoprotein (apo) A-I gene expression and decreases apoA-II gene expression in rat liver. In the present study, the mechanism of glucocorticoid action on hepatic apoA-I and apoA-II expression was studied. A single injection of rats with dexamethasone increased hepatic apoA-I mRNA levels within 6 h and further increases were observed after 12 h and 24 h. In contrast, liver apoA-II mRNA levels gradually decreased after dexamethasone treatment to less than 25% control levels after 24 h. In rat primary hepatocytes and McARH8994 hepatoma cells, addition of dexamethasone increased apoA-I mRNA levels in a time dependent and dose-dependent manner, whereas apoA-II mRNA levels were unchanged. Simultaneous addition of the glucocorticoid antagonist RU486 prevented the increase in apoA-I mRNA levels after dexamethasone treatment, which suggests that the effects of dexamethasone are mediated through the glucocorticoid receptor. Inhibition of transcription by actinomycin D and nuclear-run-on experiments in McARH8994 cells and primary hepatocytes showed that dexamethasone induced apoA-I, but not apoA-II, gene transcription. Transient-transfection assays in McARH8994 cells with a chloramphenicol acetyl transferase vector driven by the rat-apoA-I gene promoter demonstrated that the proximal apoA-I promoter could be induced by dexamethasone, and this effect could be abolished by simultaneous treatment with RU486. However, in COS-1 cells, apoA-I promoter transcription was not induced by dexamethasone or cotransfected glucocorticoid receptor. In addition, the induction of apoA-I gene transcription by dexamethasone was blocked by the protein-synthesis inhibitor cycloheximide, which suggests the presence of a labile protein involved in apoA-I gene activation by dexamethasone. In conclusion, our results demonstrate that dexamethasone regulates rat apoA-I, but not apoA-II, gene expression through direct action on the hepatocyte. The induction of apoA-I gene transcription by dexamethasone requires the glucocorticoid receptor and a labile cell-specific protein. PMID- 8706755 TI - Isolation and expression of a cDNA clone encoding human kynureninase. AB - Kynureninase (L-kynurenine hydrolase), a pyridoxal-5'-phosphate-(pyridoxal-P) dependent enzyme, catalyses the cleavage of L-kynurenine and L-3 hydroxykynurenine into anthranilic and 3-hydroxyanthranilic acids, respectively. In this report, we describe the isolation of a cDNA clone encoding human kynureninase. Degenerate oligonucleotides designed from the amino acid sequences of peptides from rat liver kynureninase, were used as primers for reverse transcription PCR of rat kidney RNA. The resulting rat cDNA product was then used to screen a human hepatoma cell line (Hep G2) cDNA library. Analysis of a positive cDNA clone showed the presence of an insert of 1651 nucleotides containing an open reading frame coding for a protein of 456 amino acids (theoretical molecular mass = 52357 Da). The predicted amino acid sequence of human kynureninase displayed high similarity to that reported for the rat enzyme and to a Saccharomyces cerevisiae gene product putatively ascribed to kynureninase. Profile analysis of kynureninase primary structure indicated the presence of a pyridoxal-P-binding site consensus sequence assigned to class-V aminotransferases, with Lys276 being the residue binding the cofactor. RNA blot analysis of human tissues, including brain, showed the presence of an approximately 2.0-kb mRNA species in all tissues tested. A second mRNA species (approximately 2.6 kb) was also detected in some tissues. After transfection of HEK-293 cells with the cDNA coding for kynureninase, the K(m) values of L kynurenine and DL-3-hydroxykynurenine for the recombinant enzyme were 671 +/- 37 microM and 13.2 +/- 2.0 microM, respectively. PMID- 8706756 TI - 4-Hydroxybenzoate hydroxylase from Pseudomonas sp. CBS3. Purification, characterization, gene cloning, sequence analysis and assignment of structural features determining the coenzyme specificity. AB - 4-Hydroxybenzoate hydroxylase from Pseudomonas sp. CBS3 was purified by five consecutive steps to apparent homogeneity. The enrichment was 50-fold with a yield of about 20%. The enzyme is a homodimeric flavoprotein monooxygenase with each 44-kDa polypeptide chain containing one FAD molecule as a rather weakly bound prosthetic group. In contrast to other 4-hydroxybenzoate hydroxylases of known primary structure, the enzyme preferred NADH over NADPH as electron donor. The pH optimum for catalysis was pH 8.0 with a maximum turnover rate around 45 degrees C. Chloride ions were inhibitory, and competitive with respect to NADH. 4 Hydroxybenzoate hydroxylase from Pseudomonas sp. CBS3 has a narrow substrate specificity. In addition to the transformation of 4-hydroxybenzoate to 3,4 dihydroxybenzoate, the enzyme converted 2-fluoro-4-hydroxybenzoate, 2-chloro-4 hydroxybenzoate, and 2,4-dihydroxybenzoate. With all aromatic substrates, no uncoupling of hydroxylation was observed. The gene encoding 4-hydroxybenzoate hydroxylase from Pseudomonas sp. CBS3 was cloned in Escherichia coli. Nucleotide sequence analysis revealed an open reading frame of 1182 bp that corresponded to a protein of 394 amino acid residues. Upstream of the pobA gene, a sequence resembling an E. coli promoter was identified, which led to constitutive expression of the cloned gene in E. coli TG1. The deduced amino acid sequence of Pseudomonas sp. CBS3 4-hydroxybenzoate hydroxylase revealed 53% identity with that of the pobA enzyme from Pseudomonas fluorescens for which a three dimensional structure is known. The active-site residues and the fingerprint sequences associated with FAD binding are strictly conserved. This and the conservation of secondary structures implies that the enzymes share a similar three-dimensional fold. Based on an isolated region of sequence divergence and site-directed mutagenesis data of 4-hydroxybenzoate hydroxylase from P. fluorescens, it is proposed that helix H2 is involved in determining the coenzyme specificity. PMID- 8706757 TI - An immobilised peptide array identifies antibodies to a discontinuous epitope in the extracellular domain of the bovine growth hormone receptor. AB - Using an array of overlapping decapeptides representing the extracellular domain of the bovine (b) growth-hormone receptor (GHR) we have mapped the continuous, dominant epitopes defined by five rabbit and one guinea pig polyclonal antisera to recombinant bovine growth-hormone-binding protein (rbGHBP). We report that six major epitopes are identified by these antisera and that these largely occur in areas of non-ordered secondary structure, although there is some contribution from the extensive beta-sheet structure of GHBP. Similar to our previously described studies for growth hormone (GH), we have again found slight differences between animals in the exact location of these epitopes. Using peptide-affinity chromatography we have isolated a population of antibodies reactive with epitope 1 (the N-terminal epitope:GHBP residues 21-38). Analysis of these antibodies by further peptide affinity chromatography and competitive radioimmunoassay experiments indicated cross-reactivity of epitope-1-specific antibodies with epitope 4 (in the interdomain hinge region of the GHBP:residues 111-126). We suggest that, although separate in the primary structure of the molecule, the tertiary fold exhibited by GHBP may bring into close proximity areas of sequence representing epitope 1 and epitope 4 such that they represent a conformational epitope. Under these conditions our experiments indicate that peptides 1 and 4 may represent partial functional epitopes for this antibody population and consequently demonstrate that this approach may be useful in describing discontinuous epitopes. PMID- 8706758 TI - Enzymatic properties and inhibition by single-stranded autonomously replicating sequences of adenylosuccinate synthase from Saccharomyces cerevisiae. AB - Adenylosuccinate synthase (ASS) from Saccharomyces cerevisiae has been shown to bind specifically to the T-rich side of the autonomously replicating sequence (ARS) core consensus sequence [Zeidler, R., Hobert, O., Johannes, L., Faulhammer, H. & Krauss, G. (1993) J. Biol. Chem. 268, 20191-20197]. We have cloned and sequenced the gene for ASS and have studied in detail the enzymatic properties and DNA-binding activity of ASS. The deduced amino acid sequence of the yeast ASS is highly similar to the same enzymes from other sources from which it is however distinguished by its more basic nature. We show that the enzymatic activity of ASS is inhibited in a highly specific manner by the binding of a 44-base DNA oligonucleotide carrying the ARS core consensus sequence. Other nucleic acids, rNTP and dNTP are not able to mimic the specific inhibitory effect. Single-base substitutions in the ARS core sequence lead to a tenfold reduction in inhibition. The inhibition data corroborate the earlier report on the DNA-binding specificity of this enzyme. The homologous enzymes from Escherichia coli and Dictyostelium discoideum do not show specific binding to single-stranded ARS sequences and their enzymatic activity is not influenced by the presence of a 44-base DNA oligonucleotide carrying the ARS core consensus sequence. Treatment of ASS with alkaline phosphatase leads to a loss of DNA binding and to a loss of the inhibition by DNA of the enzymatic activity which suggests that the DNA-binding activity but not the enzymatic activity may be regulated by the phosphorylation status of the protein. PMID- 8706759 TI - Brain capillary endothelial cells express two forms of erythropoietin receptor mRNA. AB - To study the existence of the erythropoietin receptor (Epo-R) mRNA in brain capillary endothelial cells, the reverse transcription (RT) PCR was performed using total RNAs from rat brain capillary endothelial cells (RBECs) and MBEC4, which is one of the established mouse brain capillary endothelial cell lines. Southern analysis of the RT-PCR products indicated that both RBECs and MBEC4 expressed an authentic form of Epo-R mRNA as a minor form and an intron-5 inserted form of Epo-R mRNA, thus a soluble form of Epo-R mRNA, as a major form. Furthermore, the effect of recombinant human erythropoietin (rHuEpo) on the DNA synthesis in RBECs was analyzed. rHuEpo showed a dose-dependent mitogenic action on RBECs as a competence factor. Radioiodinated rHuEpo was bound specifically to RBECs with time, cell number and dose dependencies. Binding studies with 125I rHuEpo showed that RBECs had a single class of receptors with low-affinity (Kd = 860 pM) and that the number of sites/cell (10300) was abundant. These results suggest that brain capillary endothelial cells express not only an authentic form of Epo-R but also a soluble form of Epo-R and that erythropoietin acts directly on brain capillary endothelial cells as a competence factor. PMID- 8706760 TI - Asparaginyl-tRNA synthetase from Thermus thermophilus HB8. Sequence of the gene and crystallization of the enzyme expressed in Escherichia coli. AB - The gene for the asparaginyl-tRNA synthetase, a class IIb enzyme, from the extreme thermophile Thermus thermophilus HB8 has been cloned and sequenced. Sequence analysis revealed an open reading frame that codes for a protein of 438 amino acid residues (50875 Da). Codon usage in the asparaginyl-tRNA synthetase gene (asnS) is similar to the characteristic usage in the genes for proteins from bacteria of the genus Thermus, and the G+C content in the third position of the codons is as high as 94%. The amino acid sequence of asparaginyl-tRNA synthetase from T. thermophilus shows high similarity with other bacterial asparaginyl-tRNA synthetase sequences (30-55% identity). By expression of the T. thermophilus asnS gene in Escherichia coli, the thermostable enzyme was overproduced and purified to homogeneity by heat treatment and two chromatography steps. The protein obtained is remarkably thermostable and retains 50% of its initial tRNA aminoacylation activity after 1 h of incubation at 90 degrees C or 21 h at 85 degrees C. Crystals of the enzyme were obtained from polyethylene glycol 6000 solutions by vapour diffusion techniques. The crystals diffract X-rays beyond 2.8 A. PMID- 8706761 TI - Cyclosporin A inhibits the degradation of signal sequences after processing of presecretory proteins by signal peptidase. AB - Targeting of presecretory proteins to, and insertion into, the microsomal membrane are mediated by signal sequences. These signal sequences are removed from presecretory proteins by signal peptidase. We demonstrate that the signal sequence of preprolactin, after translocation into microsomes and cleavage by signal peptidase, is converted to an intermediate form. This intermediate was found outside the microsomes, where it was degraded in the presence of cytosol. Degradation of the signal sequence of another presecretory protein, preprocecropinA, occurred even in the absence of cytosol. The immunosuppressant cyclosporin A inhibited trimming of the preprolactin signal sequence and degradation of the preprocecropinA signal sequence. We observed by cross-linking studies that cleaved signal sequences are bound to two microsomal proteins prior to degradation. PMID- 8706762 TI - Triiodothyronine regulates beta 3-adrenoceptor expression in 3T3-F442A differentiating adipocytes. AB - The effect of thyroid hormones on the beta 3-adrenoceptor expression was studied in the preadipose 3T3-F442A cell line. As assessed by molecular and pharmacological analyses, triiodothyronine addition to differentiating 3T3-F442A cells caused a 2.3-fold increase in beta 3-adrenoceptor mRNA levels, which was correlated with a parallel induction of beta 3-adrenoceptor number and of beta 3 adrenoceptor coupling to the adenylate cyclase system. Nuclear transcription experiments showed that triiodothyronine did not significantly alter the transcription rate of the beta 3-adrenoceptor gene. By contrast, the hormone increased by 36% the half-life of beta 3-adrenoceptor mRNA. Triiodothyronine exhibited a discrete effect on beta 3-adrenoceptor expression when added to mature 3T3-F442A adipocytes. This study indicates that thyroid hormones exert a differentiation-dependent and post-transcriptional regulation of beta 3 adrenoceptor expression in adipocytes. PMID- 8706763 TI - Characterization and isolation of enzymes that hydrolyze short-chain acyl-CoA in rat-liver mitochondria. AB - In this study we investigated the presence of short-chain acyl-CoA hydrolases in rat liver mitochondria. One acetyl-CoA-hydrolyzing enzyme with a molecular mass of about 48 kDa was purified to apparent homogeneity as judged by SDS/PAGE. Immunoprecipitation experiments with antibodies raised to the purified protein showed that this enzyme corresponds to a minor portion of the total mitochondrial acetyl-CoA hydrolase activity, most (about 90%) of the total activity being due to an enzyme which was labile and required Triton X-100 for its stability. Neither of these acetyl-CoA-hydrolyzing enzymes appeared to be induced by treatment of rats with di(2-ethylhexyl)phthalate, a peroxisome proliferator which mediates induction of several cytosolic and mitochondrial long-chain acyl-CoA thioesterases. In addition, an enzyme that hydrolyzed acetoacetyl-CoA was partially purified; it was induced about 3.5-fold by di(2-ethylhexyl)phthalate treatment. In conclusion, these results demonstrate that rat liver mitochondria contain several enzymes capable of hydrolyzing short-chain acyl-CoA, indicating that regulation of the metabolism of short-chain acyl-CoAs and formation of ketone bodies, could be complex. PMID- 8706764 TI - Structural analysis of the O-antigenic polysaccharide from the enteropathogenic Escherichia coli O125. AB - The structure of the polysaccharide part of the lipopolysaccharide obtained from the enteropathogenic Escherichia coli O125 has been investigated. Methylation analysis, 1H-NMR spectroscopy and 13C-NMR spectroscopy revealed that the polysaccharide is composed of repeating hexasaccharide units. Smith degradation of the native O-polysaccharide resulted in a polysaccharide with four sugar residues in the repeating unit. Information on the sequence of the native O polysaccharide and the Smith-degraded product was obtained by two-dimensional techniques, namely heteronuclear-multiple-bond-connectivity and NOESY experiments. The structure of the repeating unit of the O-polysaccharide of E. coli strain O125, which has two adjacent branch-point residues, is [sequence: see text]. PMID- 8706765 TI - Drug prescription in Italy. AB - Drug use in Italy in the period 1983-1991 has been analysed. The twenty most used substances accounted for about 20% of the sold packages. The effect of marketing of new drugs is worth noting: new and more expensive drugs are preferred even when effective, safe and less expensive alternatives are available. Prescribers have to face a highly dynamic pharmaceutical market in which 30% of substances (among the 300 most sold) changes every five years. Other than a more rational formulary, independent information and continuing education have to be considered as tools to improve a more rational prescribing. PMID- 8706766 TI - ACE-inhibitor-induced cough, an adverse drug reaction unrecognised for several years: studies in prescription-event monitoring. AB - OBJECTIVE. This study examines cough recorded in Prescription-Event Monitoring (PEM) of four ACE-inhibitors. Particular attention was paid to the study of enalapril because the drug was monitored before the causal relationship between cough and ACE-inhibitors had been widely accepted. RESULTS. Several factors which had obscured the causal relationship in the individual cases were found to be also an obstacle in PEM. For example, cough was a common and non-serious event and was under-reported in the PEM study of enalapril and the rate was not strikingly different from that recorded for other drugs. Cough induced by ACE inhibitors has several characteristics which reduce the chance of a recognisable "signal'. The original questionnaires returned from doctors in the PEM study of enalapril have been reexamined. The observation that the rate of cough diminished after enalapril had been stopped rather than increased after starting, provided the best evidence of causality, because this was not affected by many biases such as the publicity that had occurred prior to doctors participating in PEM completed later reports. PMID- 8706767 TI - Cough due to ACE inhibitors: a case-control study using automated general practice data. AB - OBJECTIVES: To determine the risk of coughing as an adverse reaction to ACE inhibitors under everyday circumstances in a large population, and to study whether this adverse effect was duration or dose dependent. DESIGN: A population based case-control study. SETTING: Ten general practices of 14 Dutch general practitioners (GP), in which all consultations, morbidity and medical interventions, including drugs prescribed, were registered over the 18 month period from 1st September, 1992 to 1st March, 1994. SUBJECTS: 1458 patients with incident coughing and up to four controls per case were obtained (total 4182 controls), matched for GP. All cases and controls were 20 years or older and had no record of respiratory infection, influenza, tuberculosis, asthma, chronic bronchitis, emphysema, congestive heart failure, sinusitis, laryngitis, haemoptysis or respiratory neoplasms during the study period. RESULTS: Cases were 2.1-times more likely than controls to have been exposed to ACE inhibitors (95% CI 1.5-3.1), but after adjustment the odds ratio was 1.4 (95% CI 0.9-2.1). The crude odds ratio for captopril was 1.3 (95% CI 0.7-2.5), for enalapril 2.6 (95% CI 1.6-4.2) and for lisinopril 2.0 (95% CI 0.5-9.3). The adjusted odds ratio for captopril was 0.9 (95% CI 0.4-1.7), for enalapril 1.7 (95% CI 1.03-2.8) and for lisinopril 1.7 (95% CI 0.4-7.9). For patients who had been on ACE inhibitor treatment for no longer than 2 months the odds ratio was 4.8 (95% CI 1.7-13.3). The odds ratio declined to 2.0 (95% CI 1.1-3.8) for those who had taken an ACE inhibitor for 2-6 months, and to 1.6 (95% CI 0.9-2.7) for those on ACE-inhibitors for more than 6 months. CONCLUSION: The risk of coughing was increased twofold among ACE inhibitor users, but the odds ratios were no longer significant after controlling for several confounding factors. The risk of developing cough due to ACE-inhibitors declines with the duration of treatment, possibly due to depletion of susceptible persons. PMID- 8706768 TI - How informed general practitioners manage mild hypertension: a survey of readers of drug bulletins in 7 countries. International Society of Drug Bulletins (ISDB). AB - OBJECTIVE: To determine whether general practitioners (GP) who are readers of independent drug bulletins can be used as an international epidemiological observatory of the criteria adopted by "well informed" doctors in various countries in the management of mild hypertension. DESIGN: Questionnaire study of GPs' diagnostic criteria for mild hypertension, routine investigation and management of patients with this diagnosis. PARTICIPANTS: 206 GPs readers of independent drug bulletins in 7 countries, comprising 95 known systematic readers of a local bulletin and 111 randomly selected regular subscribers. MAIN OUTCOME MEASURES: Response rate to the questionnaire. Diagnostic criteria, routine investigations, and treatment used for patients with mild hypertension. RESULTS: The study required two months for planning and implementation. Four countries out of eleven had a response rate < or = 50% and were excluded; the frequency of responses from other countries was 69%. The average diastolic blood pressure (DBP) considered diagnostic of mild hypertension range from 94 mm Hg (lower threshold) to 106 (upper threshold). A minority (17%) of GPs routinely request the minimum recommended laboratory tests to assess patients. GPs routinely advise non-drug measures before starting a drug. Most would not start drug treatment in patients without other risk factors and a DBP below 100 mmHg. The top first choice drugs were diuretics and beta-adrenoceptor blockers. Half of the doctors were able to quote some published guide to the management of mild hypertension, and 18% cited a relevant trial. Attitudes in diagnosing and treating mild hypertension differed widely between GPs and countries. CONCLUSIONS: GP readers of drug bulletins can be used quickly and inexpensively to assess the extent to which recommended diagnostic and therapeutic practices are accepted by "well informed" doctors. The results suggest that attitudes in managing mild hypertension vary widely among GPs and countries and differ remarkably from the recommendations of published guidelines. PMID- 8706769 TI - Haemodynamic interactions of a new calcium sensitizing drug levosimendan and captopril. AB - OBJECTIVE: Levosimendan in a new inodilator drug that sensitizes troponin C in heart muscle cells to calcium, thus improving contractility. In previous studies, a single 2 mg intravenous dose of levosimendan increased cardiac output (CO) in healthy volunteers by about 40% and decreased pulmonary capillary wedge pressure in heart failure patients by 40-50%. The aim of the present, double-blind study was to evaluate the safety of concomitant use of levosimendan and an ACE inhibiting drug. METHODS: The haemodynamic effects of levosimendan, given with or without captopril, were evaluated by using 2-dimensional echocardiography, repeated blood pressure measurements and by ambulatory ECG recordings. Twenty four male patients with stable NYHA II-III heart failure (EF < 40%) after a previous myocardial infarct were given, in randomised order, a single i.v. infusion of levosimendan or placebo. The infusions were repeated after 2 weeks treatment with upto 50 mg b.i.d. of captopril. Twelve patients received levosimendan 1 mg and twelve received 2 mg. RESULTS: Mean CO was increased from 6.0 to 6.8 1 . min-1 in patients receiving 1 mg levosimendan compared to placebo, but only from 6.3 to 6.5 1 . min-1 in patients receiving 2 mg. The increase in CO was statistically significant when all levosimendan patients were compared to placebo. Heart rate did not change after either dose. Mean stroke volume increased significantly after 1 mg but not after 2 mg of levosimendan. The addition of captopril did not change the effects of levosimendan. No additional decrease in systolic or diastolic blood pressure was observed when levosimendan and captopril were given concomitantly. CONCLUSION: It seems that concomitant treatment with captopril does not change the haemodynamic effects of levosimendan. No adverse haemodynamic interactions were seen. PMID- 8706770 TI - An assessment of the systemic effects of single and repeated doses of inhaled fluticasone propionate and inhaled budesonide in healthy volunteers. AB - The systemic effects of single and multiple doses of inhaled fluticasone propionate (FP) and budesonide were examined in 24 healthy male volunteers (age range 18-29 years). The study was of an open, placebo-controlled, randomized, three-way crossover design. On each study day, multiple blood samples were taken over a 20 h period after drug administration (after a single dose and after the last of seven doses) and area under the curve (AUC(0-20)) for plasma cortisol and white blood cell (WBC) counts was calculated. RESULTS. The present study shows that multiple dosing with FP 1.0 mg b.i.d. for 3.5 days (seven doses) resulted in a marked cortisol suppression from placebo which, at 55%, was more than double that seen with a single dose (25% suppression). Multiple dosing with budesonide 0.8 mg b.i.d. resulted in a 34% suppression in plasma cortisol compared with a suppression of 26% with a single dose. The increase in systemic activity of FP after multiple dosing is confirmed by both the number of subjects with 0800 hours plasma cortisol values below normal limits and by the changes in WBC and differential counts. CONCLUSION. The results of the present study confirm previous findings with regard to the more marked systemic effect of FP following multiple dosing as compared with a single dose. This increase in systemic effect from single dosing to multiple dosing is significantly greater for FP than for budesonide. PMID- 8706771 TI - Improved pharmacodynamics of L-asparaginase-loaded in human red blood cells. AB - To evaluate the modification of pharmacodynamic parameters induced by the administration of L-asparaginase loaded into red blood cells, 13 patients received a single dose of L-asparaginase internalised into the carrier. The enzyme was loaded using a reversible lysis-resealing process. The dose per patient ranged from 30 to 200 i.u. kg-1. Considerable heterogeneity occurred between patients. the level of L-asparaginase circulating after 24 h represented 47% of the total injected dose as compared to 74.8% for red blood cells (RBCs). However, the half-life of the enzyme remaining in the circulation was very similar to that of the RBC carrier, i.e. 29 days and 27 days, respectively, compared with 8-24 h for the free enzyme. Sustained elimination of plasma L asparagine occurred, the duration of which was dependent on the injected dose. A single injection of 30.i.u.kg-1 was sufficient to eliminate plasma L-asparagine over 10 days. With 150-200 IU.kg-1 the elimination period was extended to 50 days. These data show that the use of RBCs as carriers of L-asparaginase greatly improves the pharmacodynamic parameters of the drug. PMID- 8706772 TI - A pharmacokinetic and pharmacodynamic evaluation of buffered sublingual captopril in patients with congestive heart failure. AB - OBJECTIVE: The pharmacokinetics and pharmacodynamics of buffered sublingual captopril were assessed in patients with congestive heart failure (CHF). METHODS: The study was carried out in a randomised single-blind cross-over fashion (n = 6, 4 males and 2 females) and involved two study days, at least 7 days apart. Baseline measurements were carried out for plasma renin activity (PRA), blood pressure (B.P.) and heart rate (H.R.). Captopril (12.5 mg) was administered sublingually with dibasic potassium phosphate which maintained salivary pH at 7, or perorally with 100 ml of water. Further B.P., H.R. measurements and venous blood samples were taken over a 3 hour period post-drug administration. Blood samples were analysed for captopril and PRA levels. RESULTS: tmax after buffered sublingual administration of captopril, which ranged from 40-60 min (median = 40 min), was significantly shorter than after peroral administration (range 60-120 min, median = 90 min). Cmax was slightly greater after buffered sublingual than after peroral administration with mean values of 108.2 vs. 94.0 ng.ml-1. AUC values were similar after both routes of administration. Systolic and diastolic B.P. vs. time profiles for each administration method were significantly different i.e. sublingual administration produced an earlier reduction in B.P., however, HR did not differ significantly between the two routes. CONCLUSION: The data indicate that this novel administration method of captopril leads to an increased rate, but an unchanged extent of captopril absorption, suggesting a modest therapeutic advantage with the use of buffered sublingual captopril if a rapid reduction in blood pressure is required. PMID- 8706773 TI - Pharmacokinetics of dexamethasone in premature neonates. AB - OBJECTIVE: Dexamethasone is frequently used in premature neonates with bronchopulmonary dysplasia, however little is known about its disposition in this population. METHODS: We evaluated the pharmacokinetics of dexamethasone in 9 premature neonates with a mean gestational age of 27.3 weeks and a postnatal age of 21.8 days. RESULTS: There was a strong relationship between clearance (4.96 ml.min-1.kg-1) and gestational age ( r = 0.884). Pharmacokinetic parameters were grouped based on a gestational age of less than 27 weeks (Group I) and greater than 27 weeks (Group II). Mean clearance in group I and group II was 1.69 and 7,57 ml.min-1.kg-1, respectively. Mean distribution volume in group I and II was 1.26 and 2.19 l.kg-1, respectively. No significant relationships were noted between the disposition of dexamethasone and ventilator requirements or adverse effects. CONCLUSION: The pharmacokinetics of dexamethasone in premature neonates was related to gestational age. PMID- 8706774 TI - Therapeutic monitoring of nalbuphine: transplacental transfer and estimated pharmacokinetics in the neonate. AB - Nalbuphine, a mixed agonist-antagonist opiate, is commonly used as a systemic analgesic during labour. Recent reports of perinatal adverse effects prompted us to carry out therapeutic nalbuphine monitoring in obstetric analgesia. Because data on fetomaternal transfer are scarce and the pharmacokinetics of this drug in the neonate are largely unknown, we report data obtained from 28 parturients treated with nalbuphine either intramuscularly and/or intravenously during labour. Plasma nalbuphine levels were measured by high-performance liquid chromatography (HPLC) with electrochemical detection. At delivery, 30-150 min after maternal administration, nalbuphine concentrations ranged from 5.0 to 79.2 ng.ml-1 in mother plasma samples and from 3.0 to 46.6 ng.ml-1 in umbilical cord plasma samples. Nalbuphine concentrations were highly correlated to dose. The fetomaternal ratio was high: 0.74 and not correlated to the administered dose of nalbuphine. An estimated plasma half-life of 4.1 h was calculated from two determinations in the neonate based on the assumption of a monoexponential decay of nalbuphine concentrations. Apart from a flattening of the fetal heart rate tracing in 54% of the cases, only one neonate had a low Apgar score at birth. The apparent prolonged half-life of nalbuphine in the neonate indicates the usefulness of an intramuscular injection of naloxone to prevent recurrence of cardiorespiratory depression due to nalbuphine administration to the mother. PMID- 8706775 TI - Omeprazole and lansoprazole are not inducers of cytochrome P4501A2 under conventional therapeutic conditions. AB - OBJECTIVES: Claims that substituted benzimidazole molecules induce cytochromes P4501A2 are still controversial. This study was undertaken to evaluate their inducing potency under conventional therapeutic conditions. METHODS: Twelve healthy non-smoking young volunteers were given 20 mg omeprazole or 30 mg lansoprazole daily, in random order, for 2 weeks, separated by a 3 week wash-out period. We evaluated the CYP1A2 activity by the ratio of the molar urinary concentrations (CUM ratio) of the three end products of the paraxanthine demethylation of caffeine over the molar concentration of a paraxanthine 8 hydroxylation product. RESULTS: This urinary metabolite ratio has previously been shown to be correlated with caffeine clearance. There was slight but non significant enhancement of the CUM ratio after 2 weeks of treatment with omeprazole (3.62 (1.58) on Day 15 vs 3.09 (1.43) on Day 1), and after lansoprazole (4.26 (2.3) vs 3.65 (2.36)). Similarly, one week of treatment did not significantly alter the CUM ratio after omeprazole or lansoprazole (3.11 (1.58) and 3.28 (1.59), respectively on Day 8). CONCLUSION: The results show that both omeprazole and lansoprazole in the daily recommended therapeutic doses of 20 mg and 30 mg, respectively, have no influence on the metabolism of caffeine, and therefore no influence on cytochrome CYP1A2 activity. PMID- 8706776 TI - Pharmacokinetics of quinine in patients with chronic renal failure. AB - METHODS. We investigated the pharmacokinetics of quinine (Qn) following administration of a single oral dose of 600 mg Qn sulphate in six male Thai patients with a moderate degree of chronic renal failure (CRF), and six male Thai subjects with normal renal function. RESULTS. The drug was well tolerated in both groups of subjects; no major adverse reactions were observed. A marked alteration in the pharmacokinetics of Qn was found in patients with CRF compared to healthy subjects; there were six significant changes in the pharmacokinetic parameters. Absorption was delayed, but increased in CRF (tmax 4.5 vs 1.6 h, Cmax 6.17 vs 3.45 micrograms.ml-1). Total clearance was significantly reduced (0.94 vs 2.84 ml.min-1.kg-1, whereas Vz/f remained unchanged (1.82 vs 2.78 l. kg-1). This resulted in the increased values of AUC and prolongation of the t1/2z and MRT in the patients (AUC 181.5 vs 61.8 micrograms.min-1.ml-1, t1/2z 26 vs 9.7 h, MRT 36.4 vs 11.3 h). Median concentrations of plasma unbound fraction of Qn collected at 4 h after drug administration in patients and healthy subjects were 7.3 vs 9.8%, respectively. PMID- 8706778 TI - The anti-ulcer drug sucralfate does not affect gastric nicotine levels. AB - OBJECTIVE: It has been claimed that sucralfate can overcome the negative effects of nicotine in patients with peptic ulcer disease, although the possible mechanism being unknown. This study was performed in order to test whether sucralfate was capable of binding intragastric nicotine, thus making it impossible for the substance to exert effect. METHOD: Nicotine was administered via transdermal patches or as capsules yielding gastric concentrations of 40-2980 ng.ml-1. Gastric juice aspirates (n = 9) were incubated with sucralfate, which was then separated by centrifugation, and the nicotine concentration was compared in incubated and non-incubated samples. RESULTS: A median decrease of 13% (range 0-27%) in nicotine concentration was seen after incubation with sucralfate (P = 0.01). CONCLUSION: The binding of nicotine to the precipitating agent sucralfate is not sufficient effectively to remove nicotine from the gastric juice. PMID- 8706777 TI - Impact of quinidine on plasma and cerebrospinal fluid concentrations of codeine and morphine after codeine intake. AB - OBJECTIVE: The analgesic effect of codeine depends on its O-demethylation to morphine via sparteine oxygenase (CYP2D6) in the liver and presumably also via this enzyme in the CNS. We studied the ability of quinidine, which is a potent inhibitor of CYP2D6, to penetrate the blood brain barrier and its possible impact on codeine O-demethylation in CNS. METHODS: The study comprised 16 extensive and one poor metaboliser of sparteine, who underwent spinal anaesthesia for urinary tract surgery or examination. Eight patients were given an oral dose of 125 mg codeine and 9 patients (including the poor metaboliser) were given 200 mg quinidine 2 h before the same dose of codeine. Plasma and spinal fluid samples were collected 2 h after codeine intake. RESULTS: Free concentrations of quinidine were 11-times lower in cerebrospinal fluid than in plasma, and ranged from 9-15 nmol.l-1. Morphine concentrations were significantly lower in patients pre-treated with quinidine, both in plasma (median 1.45 nmol.l-1, range 0.74-1.95 nmol.l-1 vs 9.86 nmol.l-1, range 4.59-28.4 nmol.l-1) and in cerebrospinal fluid (0.23, 0.16-0.61 nmol.l-1 vs 3.63, 0.6-8.09 nmol.l-1). The morphine/codeine concentration ratio in plasma (3.07 x 10 (-3), 1.68-3.68 x 10 (-3) vs 19.87 x 10 (-3), 9.87-66.22 x 10 (-3) and in cerebrospinal fluid (0.83 d 10 (-3), 0.58-1.45 x 10 (-3) vs 7.19 x 10 (-3), 2.03-17.7 x 10 (-3) was also lower. The morphine/codeine concentration ratios were significantly lower in cerebrospinal fluid both without and with quinidine, but the difference between the plasma and spinal fluid ratio was significantly smaller with quinidine than without (p = 0.0002). CONCLUSION: Quinidine penetrates the blood brain barrier poorly, but quinidine pre-treatment leads to pronounced lowering of the cerebrospinal fluid concentration of morphine after codeine intake. However, the O-demethylation of codeine in CNS may not be totally blocked by quinidine. PMID- 8706779 TI - Prostaglandin E1 increases binding of 123I-low-density lipoprotein to the human liver in vivo. AB - Previous in vitro radioligand binding data have shown that prostaglandin E1 (PGE1) increases the number and the binding affinity of low-density lipoprotein (LDL) receptors of the human liver. Experimental data in normo- and hypercholesterolaemic rabbits have confirmed these findings, showing a significant increase in LDL-binding to the liver in vivo after prolonged PGE1 therapy. METHODS. This study aimed to confirm the experimental and animal data in human in vivo. 123I-LDL binding to the liver was quantified in vivo in patients suffering from peripheral vascular disease, seven of them with heterozygous familial hypercholesterolaemia (HC) and five with normal total plasma cholesterol, after PGE1 administration (5 ng.kg-1.min-1; 6 h daily for 5 days/week for 5 weeks). LDL uptake by the liver was quantified by single photon emission computer tomography (SPECT). RESULTS. The amount of LDL trapped by the liver in normocholesterolaemics (45.6%) was significantly higher than in hypercholesterolaemics (22.0%). PGE1 induced an increase in liver LDL binding, which was more pronounced in HC (+38.2%) than in normocholesterolaemic patients (+8.11%). PMID- 8706780 TI - Acute tubular necrosis following high-dose cyclosporine A therapy. PMID- 8706781 TI - Molecular markers of senescence in fibroblast-like cultures. AB - The loss of replicative capacity in vitro of normal human diploid fibroblasts is a model for studying molecular changes that accompany both regulated growth control and cellular senescence. We describe the molecular phenotype of senescent fibroblasts in terms of markers that are altered with proliferative decline. We describe these markers by analyzing pathways and associated mechanisms related to the responsiveness of proliferatively competent and senescent cells to growth signals including changes in the extracellular environment, growth factors, growth factor receptors, secondary messengers, cell-cycle progression, transcription factors, and the fidelity of DNA synthesis. There is an abundance of molecular markers for senescence in culture at every level of information transfer. Although it seems clear that some alterations in gene expression with senescence are the result of specific changes in upstream events, more global dysregulation of coordinated growth control point to as yet undefined mechanisms. PMID- 8706782 TI - Effect of age on the fatty acid composition of phospholipids in human lymphocytes. AB - We have examined the fatty acid composition of phospholipids of unstimulated and PHA-stimulated T cells from young and old donors. Our results demonstrate that aging is accompanied by decreases in the saturated fatty acids, myristic acid, and palmitic acid, and a concomitant increase in the unsaturated arachidonic acid. Following activation with PHA for 24 h, age-associated differences in fatty acids could no longer be detected. In contrast to the lymphocyte, aging did not affect the fatty acid composition of either serum or neutrophil phospholipids. Exposure of lymphocytes from old donors to myristic acid complexed medium increased the levels of myristate in the phospholipids to levels similar to that seen in lymphocytes from young donors. We conclude from these studies that aging is accompanied by an alteration in the fatty acid profiles of phospholipids, and that incubation in myristic acid complexed medium modulates these profiles. These alterations are unique to lymphocytes and may contribute to the age-related declines in lymphocyte function. PMID- 8706783 TI - A prospective view on phosphatases and replicative senescence. AB - Addition of phosphate to proteins by kinases, or its removal by phosphatases, is probably the control mechanism most often used by cells to maintain homeostasis. This mechanism presents the advantage of being fast, versatile, and easily reversible. It is used by all organisms from bacteria to man. Although more is known about the kinases, recent studies are beginning to shed light on the role of phosphatases, the enzymes that are responsible for terminating the effects of phosphorylation. These enzymes are perfect candidates for controlling all the crucial check points during cell cycle traverse, and as such, will be found to be responsible for many important decision in the life of a cell, including entry into replicative senescence. PMID- 8706784 TI - Impaired expression of hematopoietic growth factors: a candidate mechanism for the hematopoietic defect of aging. AB - Aged humans and aged experimental animals exhibit a diminished ability to upregulate hematopoiesis, but the mechanism responsible for this is unknown. The purpose of the studies was to test the hypothesis that this disregulation might be attributable to altered expression of hematopoietic growth factors. We studied the in vitro ability of cells from aged humans or mice to release bioactive colony stimulating activity (CSA) and to accumulate mRNA for defined growth factors. Human light density leukocytes from the aged released less CSA than cells from the young. GM-CSF accounted for 72-100% of the CSA released by young humans, as judged by antibody inhibition studies. In contrast, GM-CSF accounted for 0-42% of CSA from aged humans. The observation of diminished expression of GM CSF by cells from aged humans was further accompanied by a reduced accumulation of mRNA for GM-CSF. In other in vitro studies, splenic cells from aged mice released less CSA than cells from young mice, and in vivo experiments showed that splenic cells from mice challenged with E. coli accumulated less mRNA for M-CSF than young mice. These data demonstrate a similar defect in the expression of hematopoietic growth factors in aged man and mice. We propose that this diminished production of bioactive and identifiable hematopoietic growth factors may be mechanistically important in explaining the disordered hematopoiesis of aging. PMID- 8706785 TI - Expression of senescence-induced protein WS3-10 in vivo and in vitro. AB - In our efforts to characterize cellular senescence we have shown that the mRNA encoding WS3-10 protein is overexpressed in senescent human diploid fibroblasts (HDF) when compared with their younger counterparts, and that forced expression of the WS3-10 cDNA in young HDF results in suppression of calcium-dependent membrane currents, presumably due in part to the presence of a calcium binding domain within the WS3-10 protein. We have now expressed this protein in E. coli and have obtained affinity purified antibodies. Western blot analysis utilizing these antibodies showed that WS3-10 protein is also overexpressed in senescent HDF when compared to young HDF, and in normal fetal lung HDF when compared to SV40-transformed fetal lung HDF. HeLa cells do not express WS3-10 protein. In addition, we looked for WS3-10-related species in a variety of rat tissues. Analysis of WS3-10 immunologically related proteins in rat tissue extracts revealed two WS3-10 homologs, sized 22 kDa and 20 kDa. The latter presumably result from proteolytic removal of the C-terminal end of the 22 kDa polypeptide. The ratio between these polypeptides varies in a tissue-specific manner. Two proteins immunologically related to WS3-10 with sizes of 39 kDa and 91 kDa were present in rat spleen and skeletal muscle, respectively. PMID- 8706786 TI - Identification of gene sequences overexpressed in senescent and Werner syndrome human fibroblasts. AB - The phenotype of replicative senescence is a dominant trait in human diploid fibroblasts (HDF). Therefore, we have sought to identify overexpressed and/or newly expressed causal genes by constructing and screening a subtracted cDNA library derived from polyA+RNA of prematurely senescent Werner syndrome (WS) HDF. We have identified 15 cDNA clones that are overexpressed in senescent and WS HDF. Among them are six known sequences coding for: acid sphingomyelinase, fibronectin, SPARC, nm23-metastasis suppressor protein, and two translation factors, eIF-2 beta and EF-1 alpha. Among the 10 unknown clones are: S1-5, which encodes a secreted protein containing EGF-like domains and paradoxically stimulates DNA synthesis of young HDF in an autocrine and paracrine manner, S1-3, which encodes a protein containing "zinc finger" domains, suggesting nucleic acid binding properties; S1-15, which shows sequence similarities to human alpha 2 chimerin; and S2-6, which represents a new member of the LIM family of proteins. The other five clones do not have any significant homology to known sequences. Steady-state mRNA levels of all gene sequences thus far studied are elevated in both WS and senescent normal HDF when compared to young HDF, which suggests that senescent and WS HDF enter a final common pathway where multiple gene overexpression may generate diverse antiproliferative mechanisms and pathogenic sequelae. PMID- 8706787 TI - Altered expression of plasminogen activator and plasminogen activator inhibitor during cellular senescence. AB - Fibroblast senescence is associated with a loss of proliferative potential and an alteration in extracellular gene expression. Because the expression of extracellular gene products are frequently growth state dependent, we undertook a comparative study of the regulation of the components of the plasminogen activation system in young and senescent cells under controlled conditions of growth. Young and senescent cells were compared in quiescent and activated growth conditions for the secretion of tissue-type plasminogen activator (t-PA), urokinase-type plasminogen activator (u-PA), plasminogen activator inhibitor-1 (PAI-1) and plasminogen activator inhibitor-2 (PAI-2). Whereas young cells showed decreased levels of PAI-1 in the secreted and extracellular matrix pools upon serum deprivation, senescent cells showed a more constitutive pattern of gene expression, with no noticeable decrease of the levels in a low concentration of serum. RNA analysis revealed that senescent lung and skin cells, independent of the growth state, constitutively express levels of u-PA and PAI-1 comparable to the expression levels in young mitotically growing cells. These expression levels are down-regulated in quiescent young cells. In contrast, both t-PA and PAI-2 were markedly overexpressed in senescent skin lung cells under all growth conditions. Total plasminogen activator activity in conditioned medium was 50 fold higher in senescent-cell medium compared to young when cultured in 0.5% fetal calf serum (FCS) for five days, with the majority of the activity co migrating on zymograms with u-PA. Increases in PAI-1 was also observed in senescent human umbilical vein endothelial cells. In summary, cells of various types display alterations in plasminogen activator activity during replicative senescence. The inappropriate over-expression of plasminogen activator activity in vivo may be expected to lead to a progressive disruption of extracellular matrix maintenance. Thus, our observations suggest that cellular replicative senescence is associated with an altered expression of several genes regulating tissue maintenance which, in turn, could lead to degenerative changes in tissue in age-related disease(s). PMID- 8706788 TI - Epidermal growth factor and platelet-derived growth factor regulate the activity of the insulin-like growth factor I gene promoter. AB - The expression of insulin-like growth factor I (IGF-I) is regulated by hormones, oncogenes, and other growth factors, and is markedly decreased or even absent in senescent human diploid fibroblasts. In previous articles, we have reported that the SV40 large T antigen increases the production of IGF-I and that the expression of the IGF-I gene is negatively regulated by an E2F binding site in the IGF-I promoter. We have now investigated the activity of the IGF-I promoter, in response to stimulation of cells by either PDGF or EGF. Both growth factors stimulate the activity of the IGF-I promoter, indicating that they regulate the levels of expression of IGF-I. When the E2F binding sequence in the IGF-I promoter is mutated, the IGF-I promoter is constitutively active and no longer responds to the action of growth factors. PMID- 8706789 TI - In memory of Samuel Goldstein 1938-1994. Special issue. PMID- 8706790 TI - Differential effects of transforming growth factor-beta 1 on the expression of matrix metalloproteinases and tissue inhibitors of metalloproteinases in young and old human fibroblasts. AB - The balance between the activities of matrix metalloproteinases (MMPs) and the tissue inhibitors of metalloproteinases (TIMPs) is an important control point in tissue remodeling. Previous studies have demonstrated elevated expression of the MMPs collagenase and stromelysin-1 by aged human diploid fibroblasts compared to early-passage cultures. We show here that aging cells display an altered response to transforming growth factor-beta 1 (TGF beta 1) that selectively affects MMP mRNA expression. In both young and old cells, phorbol myristoyl-13 acetate (PMA) induced the expression of transcripts of collagenase, stromelysin-1, gelatinase B, TIMP-1, and TIMP-3. In young cells, TGF beta 1 reciprocally modulated PMA induced MMP and TIMP gene expression leading to reduced levels of transcripts for the MMPs and augmented accumulation of TIMP-1 and TIMP-3 mRNAs. However, repressing effects of TGF beta 1 on collagenase, stromelysin-1, and gelatinase-B RNA expression were not apparent in old cells, though induction of the TIMP genes was unimpaired. By electrophoretic mobility shift analysis the nuclear transcription factors AP1 and serum response factor (SRF) showed reduced levels of DNA binding activities in old fibroblasts compared to young cells. A probe for the TGF beta-inhibitory element (TIE) gave equivalent levels of complexes with nuclear extracts from both types of cells, though of different mobilities. We conclude that the effects of TGF beta 1 on MMP and TIMP gene expression involve different cellular intermediaries, and suggest that altered composition or modification of TIE binding factors in aging cells may underlie the failure of TGF beta 1-mediated transcription repression. This mechanism may contribute to elevated constitutive expression of MMPs in old cells and to the connective tissue deterioration that accompanies the aging process. PMID- 8706791 TI - Insights on immunological aging derived from the T lymphocyte cellular senescence model. AB - A human T cell culture system has been developed to study cellular senescence in a manner analogous to that used for fibroblasts. Using this system, several markers of replicative senescence have been identified. Evidence is provided to show that new insights into the immune deficiency of aging as well as HIV infection have emerged from the in vitro model. The culture model is now being exploited to dissect the basic mechanisms underlying the senescence program in T cells. PMID- 8706793 TI - Models of initiation of replicative senescence by loss of telomeric DNA. AB - Situated at the ends of all eukaryotic chromosomes are telomeres, genetic elements that are essential for genomic stability. It has recently been established that telomere length shortens during replicative aging of normal human somatic cells. Although the cause of replicative senescence of somatic cells is still debated, we believe that telomere shortening plays a causal role in this process. In support of this hypothesis, mutant strains of yeast and ciliates that are incapable of maintaining telomere length during cell division eventually acquire a senescent-like phenotype wherein the cells become sickly, stop growing and die. Also, replicative capacity of cultured human skin fibroblast strains shows a strong positive correlation with telomere length. Several theories explaining how telomere shortening could lead to the induction of replicative senescence are now presented. We favor a model in which replicative senescence is caused by the shortening of telomeres below a length that is critical for the maintenance of proper telomere structure and function. PMID- 8706792 TI - Proliferative cell senescence, transformation, and the recombination potential of the genome. AB - Fibroblast cell populations maintained in vitro display a different probability of escaping proliferative senescence and to transform. Observations at the cytogenetic level suggest that the long-term doubling potential of these cells is directly related to the potential for continuous chromosome rearrangements. PMID- 8706794 TI - Prohibitin and the senescent phenotype. AB - Prohibitin is an evolutionarily conserved gene that has antiproliferative activity, is ubiquitously expressed, and appears to be essential for cell survival. The gene codes for a 30 kD, post-synthetically modified protein located primarily in the mitochondria. It functionally inhibits cell cycle traverse and DNA synthesis, but its mechanism of action is presently unknown. Prohibitin is proposed to be a member of a new class of tumor suppressor genes whose inhibitory activity plays a role in the dominant senescent phenotype. Its involvement in senescence has been postulated from results obtained from such diverse systems as yeast and human diploid fibroblasts. Additional data show that prohibitin is involved in one of the limited number of pathways that results in the loss of the senescent phenotype and leads to cellular immortalization. Its involvement, however, occurs downstream in the pathway and is postulated to be part of the lost tumor suppressor activities associated with tumorigenicity. PMID- 8706795 TI - Degeneration of human oncogenes and mitochondrial genes occurs in cells that exhibit age-related pathology. AB - The development of a new class of assays to determine in vivo mutation frequencies has provided new perspectives on the timing, location, and distribution of somatic mutagenesis in mitochondrial genes and in oncogenes of the aging human body. This descriptive information has led to the inference of new models for age-related pathophysiology and oncogenesis. Mutations of mitochondrial genes rise rapidly with age to frequencies a thousand-fold higher than those of nuclear genes. Genotypic selection analysis has revealed that mitochondrial mutations accumulate predominantly in nonmitotic cells whose age dependent loss is associated with pathology. Random mitochondrial mutation is most likely to inactivate Complex I, deficiency of which induces mitochondrial superoxide formation and cell death. Genotypic selection of oncogenic mutations at the BCL2 and p53 loci has revealed that the cell specificity of oncogenic mutations in persons without cancer correlates well with sites of tumor origin, indicating that cells bearing such mutations are the likely precursors of future tumors. Quantitative variation in human BCL2 mutation frequency is extensive, and BCL2 mutation frequency rises with age, concordant with increased risk for lymphoma. The clonality and persistence of BCL2 mutations suggests two specific testable mechanisms of lymphomagenesis. BCL2 mutation frequency rises in persons exposed to cigarette smoke, and more p53 mutations occur in skin exposed to sunlight than in unexposed skin. Thus, in addition to their likely relevance to future cancer risk, the dose-response relationship between exposure and oncogenic mutations indicates promise for their future use as in vivo biodosimeters of human exposure to carcinogens. PMID- 8706796 TI - Screening for genetic predisposition to mutagens in cancer patients. AB - Both genetic and environmental factors are known to play an important role in the development of cancer. To determine whether, among individuals who develop cancers, some may have been more susceptible to the mutagenic effects of environmental agents, skin biopsies were taken from 79 cancer patients with different common types of cancers (e.g., lung, breast, bladder, colon, cervix, ovary, brain, vocal cord, uterus, skin, testis, stomach, basal cell carcinoma, leukemia, etc.). Fibroblast cultures have been established from skin explants from nearly all of the patients. The sensitivity of some of these cells as well as a number of other fibroblast strains established from "clinically normal" individuals to a battery of mutagenic agents (e.g., ethylmethane sulfonate, methylmethane sulfonate, ethidium bromide, actinomycin D, mitomycin C, bleomycin, camptothecin), which induce different kinds of DNA damage was examined. For the control group of fibroblasts, a normal range of toxicity for all of the above agents have been established. In contrast to other mutagens for which sensitivity of all of the control cell strains lay within a narrow range, large and interesting differences in sensitivity were observed for ethidium bromide. The fibroblast strains established from fetal tissue were found to be highly resistant to ethidium bromide, whereas fibroblasts from two clinically normal persons exhibited greatly enhanced sensitivity to this agent. The genetic or biochemical basis of increased sensitivity or resistance to ethidium bromide remains to be determined. The sensitivity of cells from 28 cancer patients to a number of the mutagenic agents was also examined. Most of these strains exhibited normal range of sensitivity to the mutagens; however, a few showed small but noticeable differences in sensitivity to specific agents. The fibroblast strains from cancer patients provide a useful resource to examine the genetic and metabolic factors that may be important determinants in cancer susceptibility. PMID- 8706797 TI - Replicative senescence: considerations relating to the stability of heterochromatin domains. AB - Replicative senescence of human diploid fibroblasts (HDF) cultured in vitro is characterized by a progressive and irreversible loss of responsiveness to mitogenic stimulation by serum. While some constraints have been placed on the nature of HDF senescence, its underlying molecular mechanism(s) remain obscure. Here, the possibility is considered that defects in cell cycle-coupled reassembly of repressive chromatin domains may contribute to HDF senescence. Features of this model are discussed in relation to established models of HDF senescence based on telomere shortening and loss of DNA methylation. PMID- 8706798 TI - Escape from senescence in human keratinocyte cultures. PMID- 8706799 TI - From telomere loss to p53 induction and activation of a DNA-damage pathway at senescence: the telomere loss/DNA damage model of cell aging. AB - In the cold winter of 1966 Aleksay Olovnikov, a theoretical biologist at the Academy of Sciences in Moscow, was waiting in the subway station where he was hit by the idea that the ends of linear chromosomes can't be replicated fully during each round of replication. In a theoretical paper (Olovnikov, 1971) he proposed that in somatic cells the ends of the chromosomes are not fully replicated during DNA synthesis, resulting in the shortening of linear DNA molecules with each cell division, and that this may be the cause of cell cycle arrest in senescent cells. Almost two decades after this proposal, Calvin Harley and co-workers found that telomeres, the physical ends of human chromosomes, shorten as a function of age in human cells in vitro and in vivo. The telomere hypothesis proposes that critically short telomeres may act as a mitotic clock to signal the cell cycle arrest at senescence (Harley, 1991). Here, we extend the telomere hypothesis and propose a model that incorporates recent advances in tumor suppressors and cell cycle control with several areas of cell aging. We propose that telomere shortening per se is not the direct signal for cell cycle arrest. It is the consequence of telomere loss, which may lead to generation of ds or ss DNA breaks. These breaks activate a p53 dependent or independent DNA-damage pathway that leads to the induction of a family of inhibitors of cyclin dependent kinases (including p21 and p16) and the eventual G1 block of senescence. In agreement with this hypothesis, we demonstrate that the level of p53 protein increases in near senescent cultures of MDFs. This increase may be responsible for induction of p21 (Noda, 1993) and IGF-Bp3 (Goldstein, 1991). PMID- 8706800 TI - SV40-mediated immortalization of human fibroblasts. AB - We have identified a multistep mechanism by which the DNA virus SV40 overcomes cellular senescence. Expression of SV40 T antigen is required for both transient extension of life span and unlimited life span or immortalization. These effects are mediated through inactivation of function of growth suppressors pRB and p53 via complex formation with T antigen. However, immortalization additionally requires inactivation of a novel growth suppressor gene, which has recently been identified to be on the distal portion of the long arm of chromosome 6, designated SEN6. We propose that SEN6 is responsible for cellular senescence in fibroblasts and other cells. PMID- 8706801 TI - Differential CDK-inhibitor gene expression in aging human diploid fibroblasts. AB - Cellular aging is accompanied by a reduction in proliferative activity and changes in gene expression. To further elucidate the mRNA phenotype of aging fibroblasts we have monitored the expression of an array of genes implicated in regulating cell-cycle progression. Fourteen genes, including 3 cyclin-dependent kinase (CDK) inhibitors (p16INK4, p21SDI/CIP/WAF and p27KIP), 5 cyclins, 4 CDKs, Cdi-1, and PCNA were tested in four primary fibroblast strains. Relative mRNA expression levels were assessed using a rapid and sensitive Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) assay called the "Primer-dropping" method. p16INK4, a specific inhibitor of the cyclin D-associated kinases CDK4 and CDK6, was, in addition to p21 and cyclin D1, overexpressed in higher passage cells, while the abundance of the D-type kinase mRNAs remained relatively constant. Levels of cyclin H, a component of the CDK-activating kinase (CAK) were markedly reduced in all strains examined, suggesting that the activity of target cyclin/CDK complexes may not be activated in aging cells. These results corroborate and extend previous observations demonstrating elevated expression of specific cell cycle genes in higher passage cells and suggest that overexpression of the CDK-inhibitors p16INK4 and p21SDI/CIP/WAF, but not p27KIP, may contribute to lower proliferative activity of senescing primary fibroblasts. PMID- 8706802 TI - Studies demonstrating the complexity of regulation and action of the growth inhibitory gene SDI1. AB - The identification of the DNA synthesis inhibitory gene SDI1 by investigators studying cell senescence, tumor suppression, cell cycle control and differentiation suggest a key regulatory role for this gene. To better understand the growth regulatory activity of this gene we proceeded with the experiments described here. The data demonstrate that SDI1 is an important downstream effector of p53, but here we report that it can also cause inhibition of DNA synthesis in several immortal human cell lines, independent of p53 or Rb status. Levels of SDI1 mRNA expression in immortal cells are consistently much lower than that of normal cells, indicating that immortalization is not compatible with high expression of SDI1. These results highlight the complex nature of regulation of this gene and its mode of action. PMID- 8706803 TI - Synthesis, modifications, and turnover of proteins during aging. AB - Slowing down of bulk protein synthesis is one of the most commonly observed biochemical changes during aging. The implications and consequences of slower rates of protein synthesis are manifold, including a decrease in the availability of enzymes for the maintenance, repair, and normal metabolic functioning of the cell, an inefficient removal of inactive, abnormal, and damaged macromolecules in the cell, the inefficiency of the intracellular and intercellular signalling pathways, and a decrease in the production and secretion of hormones, antibodies, neurotransmitters, and the components of the extracellular matrix. Age-related changes in the activity, specificity, and stability of a large number of proteins have been reported. However, the molecular mechanisms responsible for such alterations are still poorly understood. Studies on various components of the protein synthetic machinery have revealed a decline in the efficiency and accuracy of ribosomes, an increase in the levels of rRNA and tRNA, and a decrease in the amounts and activities of elongation factors. Because posttranslational modifications of proteins determine their activity and stability, alterations in the extent and level of various modifications such as phosphorylation, methylation, ADP-ribosylation, oxidation, glycation, and conformational changes during aging are being studied. Changes in the regulation of protein synthesis, posttranslational modifications, and protein turnover are crucial determinants of age-related decline in the maintenance, repair, and survival of the organism. PMID- 8706804 TI - Genetic modulation of the senescent phenotype of Homo sapiens. AB - The biology of aging is reviewed from the perspective of a medical geneticist. This was the perspective of the late Sam Goldstein, and this article is, therefore, dedicated to his memory. Aging can be defined as the set of phenotypes that escape the force of natural selection. These phenotypes can be modulated by mutation or polymorphism at numerous genetic loci. Given the remarkable genetic and environmental heterogeneity that characterizes our species, it is understandable that there should be considerable variation in patterns of aging. A genetic approach involving the mapping and positional cloning of major loci could provide basic understanding of the mechanisms underlying such variability. Prototypic examples being investigated by the author and his colleagues are the Werner syndrome and dementias of the Alzheimer type. The biochemical genetic analysis of these and other disorders could lead to a new style of medicine based upon preventive approaches tailored to the needs of individuals. Such interventions should ideally involve pediatricians. PMID- 8706805 TI - The role of DNA damage in cellular aging: is it time for a reassessment? AB - There is now evidence that the immediate cause of the loss of proliferative capacity in senescent cells is mediated by a specific inhibitor. If this tentative interpretation is correct, the next hurdle will be to determine mechanism(s) that regulate this putative senescence cell inhibitor that would, in effect, be the determinant of proliferative life span. One previously proposed hypothesis predicts that the decline of replicative activity is analogous to a checkpoint response to accumulated chromosomal damage (Rosenberger et al., 1991). Advances in our basic understanding of the nature of DNA damage, DNA repair mechanisms, and the response of eukaryotic cells to accumulated DNA damage provide a solid rationale for a reassessment of the causal role of the accumulation of chromosomal damage in cell senescence in vitro. PMID- 8706806 TI - Genetics of aging: current animal models. AB - Studies are summarized for three organisms-Caenorhabditis elegans, Mus musculus, and Drosophila melanogaster-utilizing three distinct approaches to the identification of longevity-determining genes: the analysis of mutations that affect life span, the use of transgenic animals to assess the effects of specific gene expression on longevity, and selective breeding to identify naturally occurring allelic variations between strains that have differential effects on life span. Correlative studies of age-dependent changes in physiology, or in cellular and molecular constituents, generally cannot discern cause from effect. In contrast, analyses of genetic influences on longevity can permit underlying mechanisms to be reliably inferred; because genotype remains essentially constant throughout life, longevity comparisons of animals differing only in genetic constitution must reflect the effects of genes on long-term survival. Understanding the genetic regulation of life span may thus lead to methods of intervention in age-associated deterioration and disease. PMID- 8706807 TI - Coming of age in culture. AB - Replicative senescence is a fundamental feature of most, if not all, somatic higher eukaryotic cells. The phenomenon has been studied for more than three decades, during which time the genetics and cell biology of senescent cells were characterized. In recent years, progress has been made on understanding the molecular basis for replicative senescence. At present, we now have a good, albeit still incomplete, understanding of some of the immediate causes for the growth arrest of senescent cells. The challenges for the future will be to understand the molecular bases for the prime causes of senescent phenotype, including the growth arrest and the altered differentiation. PMID- 8706808 TI - The evolutionary tangle of aging, sex, and reproduction and an experimental approach to its molecular dissection. AB - Exchange of genetic materials by two individual members of the same species is considered to be the origin of primitive sex. During evolution, this primitive form of molecular sex has been transformed into a complex biological function involving specialized sexual structures and multiple hormonal interactions. Development and maintenance of these reproductive structures are also dependent on hormones and hormone receptors. Furthermore, reproductive specialization in higher forms of life has led to customized species-specific rates of aging and life-span potentials that are commensurate with the reproductive needs of the particular type of organism. Because of this reproductive imposition on aging of the organism, temporal regulation of the hormone response is a significant component of the genetics of aging. We have observed a marked age-dependent alteration in the hepatic expression of the rat androgen receptor (rAR) gene. Among the large number of transcription factors that control the rAR gene, at least three appear to participate in its age-dependent regulation. Two of these are positively acting and yet/to be characterized transcription factors, while the third is a negative regulator the nuclear factor kappa B (NF-kappa B). NF kappa B is the major trans-regulator for genes involved in the immune response, inflammation, and oxidative stress. Involvement of NF-kB in the modulation of both oxidative stress and sex function provides the first example of a common molecular link between sex and aging. PMID- 8706809 TI - The age-dependent binding of CBP/tk, a CCAAT binding protein, is deregulated in transformed and immortalized mammalian cells but absent in premature aging cells. AB - CBP/tk, CCAAT Binding Protein for thymidine kinase, has been shown to bind to the distal and proximal CCAAT elements in human TK gene at G1/S boundary in normal human IMR-90 cells after serum stimulation (Pang and Chen, 1993). We now show that the serum-induced binding activity of CBP/tk was inversely related to the population doubling level (PDL) of the normal IMR-90 cells. However, little or almost no CBP/tk binding activity was observed in cells derived from patients with premature aging syndromes (e.g., Werner, Hutchinson-Gilford, and Cockayne syndrome). In contrast, CBP/tk binding activity in SV-40 virus-transformed human cells and in HeLa cells was overexpressed at levels 5- to 15-fold higher than that in normal cells and appeared to be deregulated. The half-life of CBP/tk binding activity in SV-40 transformed cells was at least 10 times longer than that in normal IMR-90 cells, suggesting that posttranslational control may contribute to the deregulation. CBP/tk binding activity detected in other mammalian cells such as murine NIH3T3, an immortal cell line, did not reveal any cell cycle dependence either. Further characterization of CBP/tk binding complex indicates that the binding complex may contain NF-YA and NF-YB and that the binding activity was sensitive to oxidizing reagents. Taken together, our data showed that the age- and cell cycle-dependent nature of CBP/tk is a function of cell types and that CBP/tk binding activity may be subjected to posttranslational and redox regulation. PMID- 8706810 TI - The regulation of trehalose metabolism in insects. AB - Trehalose is a non-reducing disaccharide comprising two glucose molecules. It is present in high concentration as the main haemolymph (blood) sugar in insects. The synthesis of trehalose in the fat body (an organ analogous in function to a combination of liver and adipose tissue in vertebrates) is stimulated by neuropeptides (hypertrehalosaemic hormones), released from the corpora cardiaca, a neurohaemal organ associated with the brain. The peptides cause a decrease in the content of fructose 2,6-biphosphate in fat body cells. Fructose 2,6 biphosphate, acting synergistically with AMP, is a potent activator of the glycolytic enzyme 6-phosphofructokinase-1 and a strong inhibitor of the gluconeogenic enzyme fructose 1,6-biphosphatase. This indicates that fructose 2,6 biphosphate is a key metabolic signal in the regulation of trehalose synthesis in insects. Trehalose is hydrolysed by trehalase (E.C. 3.2.1.28). The activity of this enzyme is regulated in flight muscle, but the mechanism by which this is achieved is unknown. Trehalase from locust flight muscle is a glycoprotein bound to membranes of the microsomal fraction. The enzyme can be activated by detergents in vitro and by short flight intervals in vivo, which indicates that changes in the membrane environment modulate trehalase activity under physiological conditions. PMID- 8706811 TI - The changing epidemiology of fungal infections: are the lipid-forms of amphotericin B an advance? AB - The incidence of invasive fungal infections is increasing and new fungal species are emerging as important pathogens. In cancer patients, the main risk factor for the development of systemic fungal disease is severe, prolonged neutropenia. Other factors, such as mucosal damage, presence of a central venous line, immunosuppressive therapy and treatment with broad spectrum antibiotics are contributory. Candida spp. are the fungi most commonly isolated in neutropenic patients. There has been a dramatic increase in non-C. albicans species, such as C. glabrata and C. krusei, largely as a result of extensive prophylactic and therapeutic use of fluconazole, to which these species are largely resistant. In neutropenic patients with candidaemia, amphotericin B is the drug of choice although the conventional formulation may be poorly tolerated. Lipid-based forms of amphotericin B, such as Abelcet, are better tolerated and can be given at a much higher dose and should therefore be considered in patients who fail on or are intolerant to the conventional agent. Aspergillosis is the second most frequent fungal infection in neutropenic patients. Primary invasive aspergillosis usually presents on chest X-ray with lung lesions and the brain is a frequent site of secondary infection. Fluconazole is inactive against Aspergillus spp. and amphotericin B is the standard treatment. Again, lipid-based forms are better tolerated than the conventional formulation in this setting, and have been shown to achieve response rates of 60% or more in a number of trials. Other potentially life-threatening fungal infections in which lipid-based amphotericin B may play an important therapeutic role in the future include cryptococcosis (increasingly problematic in AIDS patients), trichosporonosis, fusariosis and mucormycosis. Further randomized studies should be performed in a range of fungal infections to compare Abelcet with conventional amphotericin B and other lipid-based antifungal agents. PMID- 8706812 TI - Amphotericin B Lipid Complex (Abelcet) in the treatment of invasive mycoses: the North American experience. AB - Abelcet, or Amphotericin B lipid Complex, is unique formulation, comprising an equimolar mixture of amphotericin B complexed with two lipids. In preclinical studies, Abelcet was clearly demonstrated to be less toxic than amphotericin B desoxycholate and to be effective in models where amphotericin B was ineffective at its maximum tolerated dose. Pharmacokinetic studies in animals also showed that the concentration of Abelcet in blood is similar or reduced compared to levels seen with conventional amphotericin B, with accumulation in the liver, lungs and spleen. Phase I clinical trials determined the optimum tolerated dose of Abelcet to be 5 mg/kg d-1. Data are now available for 228 cases (including 51 paediatric cases) of invasive fungal infection treated with Abelcet in an open label emergency-release protocol. All patients had to have failed on previous amphotericin B or other conventional antifungals, or to have unacceptable toxicity on amphotericin B, or underlying renal disease, or nephrotoxicity due to other drugs. Abelcet was administered at a dose of 5 mg/kg d-1 for 4 wk. Approximately one-third of patients had candidiasis, one-third aspergillosis and one-third other infections, including fusariosis. Of 183 cases evaluable for response, 126 (69%) had a clinical response (cure or improvement) which was mycologically confirmed in 55% (61/110 tested). Results in paediatric cases were similar to or better than those seen in the group as a whole. When comparisons were made between cases with different types of infection, underlying disease/immunosuppressive disorder, and degree of neutropenia, the response rates were very consistent from group to group. Treatment with Abelcet was well tolerated and mean serum creatinine levels actually declined during therapy, particularly in patients with pre-existing renal dysfunction. PMID- 8706813 TI - Intensive chemotherapy and bone marrow transplantation: the challenge of fungal infections. Introduction. PMID- 8706814 TI - Meeting the challenge of systemic fungal infections in cancer: nursing implications. AB - Several factors have contributed to the increasing incidence of fungal infections over the last 2 decades, including the emergence of Acquired Immune Deficiency Syndrome, increased use of myelotoxic chemotherapy and organ transplantations, prolonged use of broad spectrum antibiotics and aggressive intensive care procedures. The two most common opportunistic fungal infections seen today are caused by Candida spp. and Aspergillus spp. Systemic fungal infections in immunocompromised patients have an extremely high mortality and require aggressive therapy. Skill in identifying early clinical features is therefore crucial and the multifaceted role of the nurse is of major importance in the management of at-risk patients. Nurses are the key resource in the prevention, early detection and treatment of fungal infection. All neutropenic patients should be thoroughly assessed at least twice a day, with special attention given to the most frequent sites of infection-the oral mucosa, lungs, skin, venepuncture sites and perineal area. Although fever is the hallmark of infection, it may be absent in the neutropenic patient who is unable to mount an adequate inflammatory response. It may also be masked by the use of certain drugs, such as steroids or analgesics. When systemic fungal infection is suspected, seriously ill patients require immediate antifungal therapy. Amphotericin B is currently the only agent with a sufficiently broad spectrum of activity to cover all the most common pathogens. Although conventional amphotericin B is effective, however, the required doses often carry significant toxicity, particularly nephrotoxicity. The new, lipid-based forms of amphotericin B, such as Abelcet, are indicated for and have been shown to be effective in patients with severe systemic and/or deep mycoses in whom conventional amphotericin B has proven ineffective or is contraindicated because of renal impairment, and in patients who have failed to respond to other antifungal agents. PMID- 8706815 TI - Palmitoylcarnitine, a surface-active metabolite. AB - Palmitoylcarnitine is a well-known intermediate in mitochondrial fatty acid oxidation. Less known are its properties as a surfactant, with a capacity to solubilize biological membranes similar to that of many synthetic detergents used in the biochemical laboratory. Some of the physico-chemical properties of palmitoylcarnitine may help to explain the need for coenzyme A-carnitine-coenzyme A acyl exchange during mitochondrial fatty acid import. The amphiphilic character of palmitoylcarnitine may also explain its proposed involvement in the pathogenesis of myocardial ischemia. PMID- 8706816 TI - A possible role for CYP27 as a major renal mitochondrial 25-hydroxyvitamin D3 1 alpha-hydroxylase. AB - A mitochondrial cytochrome P450 fraction catalyzing 1 alpha- and 27-hydroxylation but not 24-hydroxylation of 25-hydroxyvitamin D3 was purified from pig kidney. The ratio between the 1 alpha- and 27-hydroxylase activities was the same in all purification steps including a side fraction. Attempts to separate the 1 alpha- and 27-hydroxylase activities were unsuccessful. A monoclonal antibody directed against purified pig liver CYP27 recognized a protein of the same apparent M(r) and immunoprecipitated both the 1 alpha- and 27-hydroxylase activities towards 25 hydroxyvitamin D3 in the purified kidney enzyme fraction as well as in a solubilized, crude cytochrome P450 extract considered to represent the major part of the 25-hydroxyvitamin D3 hydroxylases in kidney mitochondria. Taken together, the results from the purification and the experiments with CYP27 antibody, substrate inhibition, and recombinant expressed human liver CYP27 strongly indicate that CYP27 is able to catalyze 1 alpha-hydroxylation but not 24 hydroxylation of 25-hydroxyvitamin D3 in kidney. In conclusion, the results provide evidence for a role for CYP27 as a major renal mitochondrial 25 hydroxyvitamin D3 1 alpha-hydroxylase. PMID- 8706817 TI - Site-directed mutagenesis of the formate dehydrogenase active centre: role of the His332-Gln313 pair in enzyme catalysis. AB - Gln313 and His332 residues in the active centre of NAD(+)-dependent formate dehydrogenase (EC 1.2.1.2, FDH) from the bacterium Pseudomonas sp. 101 are conserved in all FDHs and are equivalent to the glutamate-histidine pair in active sites of D-specific 2-hydroxyacid dehydrogenases. Two mutants of formate dehydrogenase from Pseudomonas sp. 101, Gln313Glu and His332Phe, have been obtained and characterised. The Gln313Glu mutation shifts the pK of the group controlling formate binding from less than 5.5 in wild-type enzyme to 7.6 thus indicating that Gln313 is essential for the broad pH affinity profile towards substrate. His332Phe mutation leads to a complete loss of enzyme activity. The His332Phe mutant is still able to bind coenzyme but not substrate or analogues. The role of histidine in the active centre of FDH is discussed. The protonation state of His332 is not critical for catalysis but vital for substrate binding. A partial positive charge on the histidine imidazole, required for substrate binding, is provided via tight H-bond to the Gln313 carboxamide. PMID- 8706818 TI - Characterization of recombinant human HBP/CAP37/azurocidin, a pleiotropic mediator of inflammation-enhancing LPS-induced cytokine release from monocytes. AB - Neutrophil-derived heparin-binding protein (HBP) is a strong chemoattractant for monocytes. We report here for the first time the expression of recombinant HBP. A baculovirus containing the human HBP cDNA mediated in insect cells the secretion of a 7-residue N-terminally extended HBP form (pro-HBP). Deletion of the pro peptide-encoding cDNA sequence resulted in correctly processed HBP at the N terminus. Electrospray mass spectrum analysis of recombinant HBP yielded a molecular weight of 27.237 +/- 3 amu. Consistent with this mass is a HBP form of 225 amino acids (mature part +3 amino acid C-terminal extension). The biological activity of recombinant HBP was confirmed by its chemotactic action towards monocytes. Furthermore, we have shown that recombinant HBP stimulates in a dose dependent manner the lipopolysaccharide (LPS)-induced cytokine release from human monocytes. PMID- 8706819 TI - Identification of an Arabidopsis thaliana cDNA encoding a HSP70-related protein belonging to the HSP110/SSE1 subfamily. AB - Heat-shock protein 70 (HSP70)-related proteins are classified in two main subfamilies: the DnaK subfamily and the HSP110/SSE1 subfamily. We have characterized the first plant member of the HSP110/SSE1 subfamily, HSP91. At least two, tightly linked genes encoding HSP91 are present per haploid Arabidopsis genome. HSP91 is constitutively expressed in non-stressed Arabidopsis plants and is transiently induced by heat shock. PMID- 8706820 TI - Uncleaved env gp160 of human immunodeficiency virus type 1 is degraded within the Golgi apparatus but not lysosomes in COS-1 cells. AB - The fate of newly synthesized human immunodeficiency virus type 1 env gp160 was examined in COS-1 cells. The results of morphological chase experiments involving cycloheximide demonstrated that gp160 was retained in the Golgi apparatus for longer than the half-life of the molecule. The degradation of gp160 was insensitive to both bafilomycin A1 and leupeptin (< 0.2 mM), which block lysosomal proteolysis. However, degradation was effectively suppressed by leupeptin at higher concentrations, maximally at 1.7 mM. Furthermore, undegraded gp160 was accumulated in the Golgi apparatus, but was not detected in lysosomes. These results indicate that in COS-1 cells gp160 is not degraded in lysosomes, but rather that degradation takes place in the Golgi apparatus. PMID- 8706821 TI - The putatively protective Onchocerca volvulus neuronal protein E1 is a member of the death domain protein family. AB - Here we show that E1, an ankyrin-related, potentially protective, neuronal protein of the human filarial nematode Onchocerca volvulus contains a death domain (DD), most similar to that of human Mort1/FADD (39% identity). In addition, sequence comparison of E1 to its homologue from Litomosoides sigmodontis and to Caenorhabditis elegans ankyrin defines two further putative functional domains. One represents the end of the spectrin-binding domain of ankyrins, the other an unique domain, most highly conserved between these nematodes, containing a calpain sequence motif. Thus, E1 may be involved in apoptosis, raising the possibility that protection against this parasitic helminth may be induced by apoptotic processes. PMID- 8706822 TI - Human erythrocyte metabolism is modulated by the O2-linked transition of hemoglobin. AB - The metabolic behaviour of human erythrocytes has been investigated with particular regard to the effect of their oxygenation state. Experiments performed at high phosphate concentration (80 mM) within the pH range 7.0-7.8 on erythrocytes at high (HOS) and low (LOS) oxygen saturation showed that at any pH value: (1) glucose consumption was independent of the oxygenation state; (2) pentose phosphate pathway (PPP) flux was about 2 times higher in the HOS than in the LOS state. At low phosphate concentration (1.0 mM) the PPP flux doubled in HOS as well as in LOS erythrocytes, whereas the decrease in glucose consumption was more marked in the HOS state. Metabolism of LOS erythrocytes approached that of HOS erythrocytes under the following conditions: (1) erythrocytes having band 3 modified by 4,4'-diisothiocyanatostilbene-2,2'-disulphonic acid; (2) CO saturated erythrocytes. These data support the hypothesis of a modulation of the relative rates of PPP and glycolysis achieved through competition between deoxy hemoglobin (deoxy-Hb) and glycolytic enzymes for the cytoplasmic domain of band 3. PMID- 8706823 TI - Differential expression of cGMP-inhibited cyclic nucleotide phosphodiesterases in human hepatoma cell lines. AB - PDE3 or cGMP-inhibited cyclic nucleotide phosphodiesterase (cGI PDE) activity was detected in homogenates of HepG2, Hep3B and HuH7, but not SK-Hep-1, human hepatoma cells. In HepG2 and Hep3B cells PDE3 activity was found predominantly in particulate fractions; in HuH7, in both particulate and supernatant fractions. cDNAs encoding two human PDE3s (an 'adipocyte' type, HcGIP1, and a 'cardiovascular' type, HcGIP2) have been cloned. HcGIP1 cDNA hybridized strongly with poly(A)+ RNA species from HepG2 and Hep3B. Both HcGIP1 and HcGIP2 mRNAs were expressed in Hep3B and HuH7 cells. The nucleotide sequence of an approximately 300-bp cDNA fragment, isolated after RT-PCR cloning from HepG2 RNA, was identical to a sequence within the conserved domain of HcGIP1 cDNA, consistent with the presence of HcGIP1 mRNA in HepG2 cells. PMID- 8706824 TI - Transmembrane topology of Escherichia coli H(+)-ATPase (ATP synthase) subunit a. AB - Escherichia coli H(+)-ATPase subunit a is a hydrophobic F0 subunit. To investigate the topology of the subunit in the membrane, we prepared site specific polyclonal antibodies against amino-terminal (Ser-3 to Leu-16), middle loop (Lys-167 to Gln-181), and carboxyl-terminal (Thr-259 to His-271) peptide segments. Enzyme-linked immunosorbent assay revealed that these antibodies specifically reacted with subunit a of inside-out membrane vesicles, but not with that of right-side-out spheroplasts. Full reactivity appeared when spheroplasts were disrupted with Triton X-100 (0.5%) or by sonication. These results suggest that at least parts of the three peptide segments of subunit a face the cytoplasm. Based on these observations, we propose a novel transmembrane topology of subunit a. PMID- 8706825 TI - Cloning and expression of human mitochondrial deoxyguanosine kinase cDNA. AB - Mammalian mitochondrial deoxyguanosine kinase (dGK) is responsible for phosphorylation of purine deoxyribonucleosides in the mitochondrial matrix. Using a RT-PCR-generated probe, based on amino acid sequence information from proteolytic fragments of purified bovine dGK, we have cloned a cDNA from a human brain cDNA library that encodes a 30 kDa protein. The deduced amino acid sequence of this protein included the sequence of all six peptides isolated and sequenced from purified dGK. Expression and purification of recombinant protein from induced Escherichia coli extracts revealed that it catalyses efficient phosphorylation of dGuo, arabinosyl guanine, dAdo, 2-chloro-2'-deoxyadenosine and dIno similar to purified dGK. Northern blot analysis demonstrated one dominant positive mRNA of 1.35 kb and it was found in several tissues at similar levels. The coding sequence of dGK showed 46% identity to the coding sequence of cytosolic deoxycytidine kinase, and conserved sequence motifs among the known deoxynucleoside kinase were identified. PMID- 8706826 TI - Identification of DNA binding-site preferences for nuclear factor I-A. AB - Nuclear factor I (NFI) proteins constitute a large family of DNA binding proteins. These proteins promote the initiation of adenovirus replication and regulate the transcription of viral and cellular genes. The binding sites for NFI have been reported in a wide variety of promoters, and they exhibit flexibility in their sequences. To clarify the DNA binding site of NFI-A, one of the NFI proteins, we performed a polymerase chain reaction-mediated random site selection, and determined the optimal sequence as 5'-TTGGCANNNN(G/T)CCA(G/A)-3'. PMID- 8706827 TI - Selective uptake and degradation of c-Fos and v-Fos by rat liver lysosomes. AB - The transcription factor c-Fos is a short-lived protein and calpains and ubiquitin-dependent systems have been proposed to be involved in its degradation. In this report, we consider a lysosomal degradation pathway for c-Fos. Using a cell-free assay, we have found that freshly isolated lysosomes can take up and degrade c-Fos with high efficiency. v-Fos, the oncogenic counterpart of c-Fos, can also be taken up by lysosomes, yet the amount of incorporated protein is much lower. c-Fos uptake is independent of its phosphorylation state but it appears to be regulated by dimerization with differentially phosphorylated forms of c-Jun, while v-Fos escapes this regulation. Moreover, we show that c-Fos is immunologically detected in lysosomes isolated from the liver of rats treated with the protease inhibitor leupeptin. Altogether, these results suggest that lysosomes can also participate in the selective degradation of c-Fos in rat liver. PMID- 8706828 TI - The construction and characterization of an effective transpositional system based on IS30. AB - We constructed an in vivo system to detect transpositional rearrangements induced by the insertion sequence IS30. The transposase protein expressed from the transposase producer plasmids catalyzed rearrangements on different target sequences presented in trans. High yields, up to 83%, of transpositional frequencies were observed. The frequency of rearrangements correlated with the amount of transposase protein produced and the attractivity of the target sequences. Alteration in the frequency of transposition was observed in the recA- E. coli strains JM109 and TG2. Remarkable structural and functional analogy was found with site-specific recombination systems. PMID- 8706829 TI - Structural and functional characterization of cytochrome c3 from D. desulfuricans ATCC 27774 by 1H-NMR. AB - Cooperativity between redox and protonation centres is known to be crucial for the function of complex proteins, but it is often difficult to describe in terms of thermodynamic parameters. Cytochrome c3 is a good model for these studies since, while retaining the overall complexity of larger systems, it is suitable for detailed crystallographic and spectroscopic studies. Assignment of the haem substituent NMR resonances, together with NMR redox titrations of cytochrome c3 from D. desulfuricans ATCC 27774, was used to correlate relative redox potentials to specific haems in the structure: haem II approximately equal to haem I < haem IV < haem III. This order is different from that determined for the homologous proteins studied and in disagreement with that previously reported for this cytochrome (Morais, J., Palma, N., Frazao, C., Caldeira, J., LeGall, J., Moura, I., Moura, J.J.G. and Carrondo, M.A. (1995) Biochemistry 34, 12830-12841). PMID- 8706830 TI - A model for DNA polymerase translocation: worm-like movement of DNA within the binding cleft. AB - On the basis of recent results, we propose a model for DNA polymerase translocation along DNA. Human immunodeficiency virus reverse transcriptase is taken as an example. According to the model, movement of the enzyme is the result of transition of the enzyme-bound DNA from the A- to B-form which is accompanied by lengthening of DNA within the binding channel. The driving force of this transition is the increase in water accessibility to the DNA-binding cleft after dNTP binding. dNTP hydrolysis proceeding during the following chemical step supplies the energy for the reverse B-->A transition of DNA. Translocation is considered to be an integral part of the stage of conformational change preceding catalysis and can be described as a worm-like movement of DNA within the DNA binding cleft. PMID- 8706831 TI - The weaver mutation changes the ion selectivity of the affected inwardly rectifying potassium channel GIRK2. AB - The weaver mutation in mice has recently been identified as a single base-pair mutation in the Girk2 gene, which encodes a G-protein-activated inwardly rectifying potassium channel, GIRK2. The mutation results in a Gly to Ser substitution at residue 156, in the putative pore-forming region of the potassium channel. In the present study, we used Xenopus oocytes to express mutant GIRK2, and to characterize the effects of the mutation on the channel. The mutation results in a loss of the normal high selectivity for K+ over Na+, with little effect on other channel properties such as activation by the mu opioid receptor. The resulting increase in basal Na+ permeability causes a marked depolarization of oocytes expressing the mutant GIRK2 protein. This result was observed even when the mutant GIRK2 was coexpressed with GIRK1, a situation more analogous to that seen in vivo. Thus, the increased Na+ permeability and resulting depolarization may contribute to the pathology of cerebellar granule cells and substantia nigra dopaminergic neurons observed in the weaver mice. PMID- 8706832 TI - GTPase properties of the interferon-induced human guanylate-binding protein 2. AB - Guanylate-binding proteins (GBPs) were originally described as proteins that are strongly induced by interferons and are capable of binding to agarose-immobilized guanine nucleotides. hGBP1, the first of two members of this protein family in humans, was recently shown to represent a novel type of GTPase that hydrolyzes GTP predominantly to GMP. We now report that purified recombinant hGBP2 also hydrolyzes GTP very efficiently, although GDP rather than GMP was the major reaction product. The biochemical parameters of this reaction were as follows: Km = 313 microM, turnover number = 22 min-1. Both hGBP1 and hGBP2 failed to hydrolyze GDP, however, GDP was an effective inhibitor of the hGBP2- but not the hGBP1-catalyzed GTP hydrolysis reaction. Thus, hGBP1 and hGBP2 have similar biochemical properties, but show pronounced differences in product specificity. PMID- 8706833 TI - Role of the N- and C-termini of porin in import into the outer membrane of Neurospora mitochondria. AB - The signals for targeting and assembly of porin, a protein of the mitochondrial outer membrane, have not been clearly defined. Targeting information has been hypothesized to be contained in the N-terminus, which may form an amphipathic alpha-helix, and in the C-terminal portion of the protein. Here, the role of the extreme N- and C-termini of porin from Neurospora crassa in its import into the mitochondrial outer membrane was investigated. Deletion mutants were constructed which lacked the N-terminal 12 or 20 residues or the C-terminal 15 residues. The porins truncated at their N-termini were imported in a receptor-dependent manner into the outer membrane of isolated mitochondria. When integrated into the outer membrane, these preproteins displayed an increased sensitivity to protease as compared to wild-type porin. In contrast, mutant porin truncated at its C terminus did not acquire protease resistance upon incubation with mitochondria. Thus, unlike most other mitochondrial preproteins, porin appears to contain important targeting and/or assembly information at its C-terminus, rather than at the N-terminus. PMID- 8706834 TI - An electrophysiological study of calcium entry during normal human T-lymphocyte activation. AB - Our aim was to observe whether normal human T-cells respond to mitogenic stimulation with large whole-cell inward currents (composed of identifiable single-channel contributions) when [Ca2+]i is not markedly lowered but instead kept normal or moderately low, as has been reported in human leukaemic Jurkat T cell line and T-cell clones [Kuno et al. (1986) Nature 323, 269-73; Kuno and Gardner (1987) Nature 326, 301-304; Gardner (1990) Annu. Rev. Immunol. 8, 231 252]. Whole-cell patch recordings showed no such currents in cells otherwise normally responding to depolarisation with the macroscopic IK described in T lymphocytes and thus deemed viable, in agreement with the notion that Ca2+ influx in normal T-cells enterily depends on depletion of internal stores [Putney (1986) Cell Calcium 7, 1-12; Putney (1990) Cell Calcium 11, 611-624]. PMID- 8706835 TI - Characterization of a new peptide from Tityus serrulatus scorpion venom which is a ligand of the apamin-binding site. AB - A new ligand (Ts kappa) of the apamin binding site on rat brain synaptosomes (K0.5 = 300 pM) was purified and characterized from the venom of Tityus serrulatus. It is a polypeptide toxin of 35 amino acid residues, with three disulfide bridges. Its cDNA was amplified from a venom gland cDNA library and the nucleotide sequence determined. A model of Ts kappa was constructed by amino acid replacement using charybdotoxin structure as determined by 1H nuclear magnetic resonance as starting model. PMID- 8706836 TI - Characterization of an Arabidopsis thaliana cDNA encoding an S-adenosylmethionine sensitive threonine synthase. Threonine synthase from higher plants. AB - An Arabidopsis thaliana cDNA encoding an S-adenosylmethionine-sensitive threonine synthase (EC 4.2.99.2) has been isolated by functional complementation of an Escherichia coli mutant devoid of threonine synthase activity. Threonine synthase from A. thaliana was shown to be synthesized with a transit peptide. The recombinant protein is activated by S-adenosylmethionine in the same range as the plant threonine synthase and evidence is presented for an involvement of the N terminal part of the mature enzyme in the sensitivity to S-adenosylmethionine. PMID- 8706837 TI - Prevention of necrosis and activation of apoptosis in oxidatively injured human myeloid leukemia U937 cells. AB - A 3 h exposure to 1 mM H2O2 followed by 6 h post-challenge growth in peroxide free medium induces necrosis in U937 cells. Addition of the poly(ADP ribose)polymerase inhibitor 3-aminobenzamide during recovery prevents necrosis and triggers apoptosis, as shown by the appearance of apoptotic bodies, extensive blebbing and formation of multimeric DNA fragments as well as 50 kb double stranded DNA fragments. Thus, the same initial damage can be a triggering event for both apoptotic and necrotic cell death. Furthermore, necrosis does not appear to be a passive response to overwhelming damage. PMID- 8706839 TI - The size differences among mammalian introns are due to the accumulation of small deletions. AB - In order to investigate the molecular mechanisms that alter intron size, we conducted an extensive interspecies comparison of homologous introns among three mammalian groups: human, artiodactyls, and rodents. The size differences of introns were statistically significant among all three groups (longest intron was for human and shortest for rodents), and appear to be due to the accumulation of small deletions, according to the separate count of insertion and deletion frequencies. The distribution of intron size differences also has a shape similar to that for the distribution of insertion/deletion sizes found in pseudogenes. It is suggested that introns are selectively neutral to small-scale changes of the genome size, which inherently contain the bias of favoring short deletions against short insertions. PMID- 8706838 TI - Antimicrobial activity of a 13 amino acid tryptophan-rich peptide derived from a putative porcine precursor protein of a novel family of antibacterial peptides. AB - It has long been speculated that porcine cathelin is an N-terminal fragment of a longer precursor protein which possesses antimicrobial activity. In an attempt to find such a precursor, a cDNA clone was recently isolated and sequenced by screening a cDNA library from porcine bone marrow. In order to identify the functional activity of the putative protein encoded by an open reading frame, we have synthesized various lengths of peptides that correspond to the C-terminal region of the protein and examined them for their antimicrobial activities. We found that a 13 amino acid tryptophan-rich region with the sequence of VRRFPWWWPFLRR had strong antimicrobial activity with a wide spectrum. It showed potency against Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumonia, Staphylococcus epidermidis, Proteus mirabilis, and Streptococcus group D as well as Aspergillus fumigatus. The action of this peptide is bactericidal rather than bacteriostatic and this activity is completely inhibited by 2 mM MgCl2. Our results indicate that the previously identified putative precursor encoded by the isolated cDNA indeed possesses a potent antimicrobial activity and that this 13 amino acid synthetic peptide is considered to be a potentially effective drug against various infectious agents. PMID- 8706840 TI - The emergence of major cellular processes in evolution. AB - The phylogenetic distribution of divergently related protein families into the three domains of life (archaea, bacteria and eukaryotes) can signify the presence or absence of entire cellular processes in these domains and their ancestors. We can thus study the emergence of the major transitions during cellular evolution, and resolve some of the controversies surrounding the evolutionary status of archaea and the origins of the eukaryotic cell. In view of the ongoing projects that sequence the complete genomes of several Archaea, this work forms a testable prediction when the genome sequences become available. Using the presence of the protein families as taxonomic traits, and linking them to biochemical pathways, we are able to reason about the presence of the corresponding cellular processes in the last universal ancestor of contemporary cells. The analysis shows that metabolism was already a complex network of reactions which included amino acid, nucleotide, fatty acid, sugar and coenzyme metabolism. In addition, genetic processes such as translation are conserved and close to the original form. However, other processes such as DNA replication and repair or transcription are exceptional and seem to be associated with the structural changes that drove eukaryotes and bacteria away from their common ancestor. There are two major hypotheses in the present work: first, that archaea are probably closer to the last universal ancestor than any other extant life form, and second, that the major cellular processes were in place before the major splitting. The last universal ancestor had metabolism and translation very similar to the contemporary ones, while having an operonic genome organization and archaean-like transcription. Evidently, all cells today contain remnants of the primordial genome of the last universal ancestor. PMID- 8706841 TI - A mechanism for beta-amyloid overproduction in Alzheimer's disease: precursor independent generation of beta-amyloid via antisense RNA-primed mRNA synthesis. AB - The overproduction of beta-amyloid (A beta) appears to be a primary cause of Alzheimer's disease (AD). A beta can be generated by proteolytic cleavage of precursor protein (beta APP) at beta- and gamma-secretase sites in both disease and normal cells. There is, however, no evidence that proteolytic processing of beta APP in sporadic AD-affected tissues differs qualitatively or quantitatively from that occurring in normal cells, and additional pathways for the enhanced production of A beta in sporadic AD which constitutes the majority of all AD cases should be considered. The major factor limiting the production of A beta in normal cells is cleavage at the alpha-secretase site within the A beta sequence. But, whereas the intact beta APP is a substrate for cleavage at the alpha secretase site, the immediate precursor of A beta, 12-kDa C-terminal beta APP fragment, is not susceptible to the alpha-secretase cleavage but it can be cleaved by gamma-secretase thus generating A beta. Moreover, the gamma-secretase cleavage is not the rate-limiting step in the production of A beta. Therefore, the increase in production of the 12-kDa C-terminal beta APP fragment may be an efficient way to overproduce A beta. A mechanism for the generation of the 12-kDa fragment independently of beta APP is proposed. It postulates an additional step of amplification of mRNA, namely the antisense RNA-mediated generation of a truncated mRNA encoding 12-kDa C-terminal fragment. Initiation of translation at the first AUG in the truncated mRNA results in a polypeptide that is cleaved by gamma-secretase generating A beta. The proposed model makes several verifiable predictions and suggests new directions of experimentation that may lead to a better understanding of the mechanisms involved in AD. PMID- 8706842 TI - Structure-activity relationships in the peptide antibiotic nisin: antibacterial activity of fragments of nisin. AB - The post-translationally modified peptide antibiotic nisin has been cleaved by a number of proteases and the fragments produced purified, characterised chemically, and assayed for activity in inhibiting the growth of Lactococcus lactis MG1614 and Micrococcus luteus NCDO8166. These results provide information on the importance of different parts of the nisin molecule for its growth inhibition activity. Removal of the C-terminal five residues leads to approximately a 10-fold decrease in potency, while removal of a further nine residues, encompassing two of the lanthionine rings, leads to a 100-fold decrease. There are some differences between analogous fragments of nisin and subtilin, suggesting possible subtle differences in mode of action. Cleavage within, or removal of, lanthionine ring C essentially abolishes the activity of nisin. The fragment nisin1-12 is inactive itself, and specifically antagonises the growth-inhibitory action of nisin. These results are discussed in terms of current models for the mechanism of action of nisin. PMID- 8706843 TI - DNA initiates polymorphic structural transitions in lecithin. AB - The inverted micellar phase, obtained by treating lecithin and Ca(2+)-DNA complex with chloroform, was used as an intermediate step in the preparation of DNA Ca(2+)-lecithin complex. DSC analysis demonstrated the involvement of a large fraction of lipid in the interaction with DNA. Freeze-fracture electron microscopy revealed (i) rod-like structures on the hydrophobic fracture surface of membranes and (ii) regular bundles of fibrils with a repeat distance of about 6 nm, which were located free in solution. Similar regular bundles of fibrils were also revealed by staining the samples with uranyl acetate. According to the suggested model, the observed structures are hexagonally packed inverted lipid tubes, with DNA located in their central cores. The possible biological relevance of the capability of Ca(2+)-DNA to initiate polymorphic phase transitions of lecithin is discussed. PMID- 8706844 TI - Topological organization of subunits VII and VIII in the ubiquinol-cytochrome c oxidoreductase of Saccharomyces cerevisiae. AB - To determine the topology of subunit VIII of the yeast ubiquinol-cytochrome c oxidoreductase in the mitochondrial inner membrane, an epitope has been introduced in the N-terminal half of this protein. Previous topology studies had shown that at least the C-terminus faces the intermembrane space [Hemrika and Berden (1990) Eur. J. Biochem. 192, 761-765]. Based on sensitivity of the protein to proteinase K digestion we now suggest that the N-terminus of subunit VIII is similarly oriented, implying that this subunit does not span the membrane. Despite this, however, subunit VIII cannot be extracted from the membrane even after treatment with 0.1 M Na2CO3 at pH 11.5, showing that the protein is integrally embedded in the membrane. A similar behaviour was displayed by another low molecular weight protein of the complex, subunit VII, which faces the matrix side. A model for the topology of these subunits in the membrane is discussed with respect to the structure of the complex and their involvement in quinone binding. PMID- 8706845 TI - N-(2-ferrocene-ethyl)maleimide: a new electroactive sulphydryl-specific reagent for cysteine-containing peptides and proteins. AB - We report the synthesis and application of a specific electroactive label, N-(2 ferrocene-ethyl)maleimide, which provides new redox properties to organic compounds and proteins possessing sulphydryl groups. Its reaction conditions with the cysteine-containing peptide, glutathione, and a terminal monooxygenase enzyme, cytochrome P450cam are presented. The labelled peptide and enzyme acquired reversible electrochemical properties due to the attached ferrocene moiety. PMID- 8706846 TI - Enzyme from the medicinal leech (Hirudo medicinalis) that specifically splits endo-epsilon(-gamma-Glu)-Lys isopeptide bonds: cDNA cloning and protein primary structure. AB - Earlier we detected a novel enzymatic activity in salivary gland secretion of the medicinal leech, splitting isopeptide bonds between the glutamine gamma carboxamide and lysine epsilon-amino group. This activity is due to destabilase. We described its partial amino acid sequence and sequences to two closely related cDNAs, but none of them perfectly matched the protein isolated. Here we report the isolation and sequence peculiarities of the third cDNA of the family as well as the complete sequence of the destabilase protein. The inferred mature protein product of this cDNA matches the independently determined destabilase protein sequence. It contains 115 amino acid residues including 14 highly conserved Cys residues and is formed from a precursor containing specific leader peptide. PMID- 8706847 TI - Thimerosal modulates the agonist-specific cytosolic Ca2+ oscillatory patterns in single pancreatic acinar cells of mouse. AB - Modulation of the agonist-specific cytosolic Ca2+ oscillatory pattern by thimerosal has been investigated in single pancreatic acinar cells using patch clamp perforated whole-cell recording to measure the calcium-dependent chloride current (I(C1)(Ca2+)). 1 microM thimerosal, which fails to evoke Ca2+ oscillation alone, clearly changed the pattern of Ca2+ oscillation from pulsatile spikes (evoked by low concentrations of activators) to sinusoidal or transient oscillations. The mimetic action of thimerosal was independent of extracellular Ca2+, was blocked by extracellular application of dithiothreitol or 10 mM caffeine, as well as by internal perfusion with heparin; but was unaffected by ruthenium red. We conclude that thimerosal modulates the agonist-specific cytosolic Ca2+ oscillatory patterns mediated by sensitizing the InsP3-induced Ca2+ release. PMID- 8706848 TI - Nuclear DNA strand breaks during ethanol-induced oxidative stress in rat brain. AB - Free radical-mediated oxidative damage has been implicated in the pathophysiological mechanisms of apoptosis. In this study we report that statistically significant strand breaks were induced primarily in the hippocampus and cerebellum during chronic, and not acute, ethanol treatment. Damage to DNA observed in hippocampus and cerebellum was also correlated with significant modification in the activities of mitochondrial respiratory complexes I and IV and with a significant increase in lipid peroxidation products. This finding lends support to the fact that hippocampus and cerebellum are brain areas particularly vulnerable to redox changes induced by alcohol intoxication, suggesting lower threshold levels of ethanol tolerance. PMID- 8706849 TI - Determination of relative abundance of splicing variants of Oreochromis glutamate receptors by quantitative reverse-transcriptase PCR. AB - In this study, the relative abundance of splicing variants of Oreochromis non NMDA subtype glutamate receptors was studied by quantitative reverse transcriptase PCR (RT-PCR). The relative expression level between the flip and flop transcripts of fGluR2 alpha determined by quantitative RT-PCR is apparently much higher than that estimated by sequence analysis of the cloned RT-PCR products. Control studies were performed to demonstrate the accuracy of the application of quantitative RT-PCR analysis in studying the relative abundance between the flip and flop transcripts of glutamate receptors. PMID- 8706850 TI - In situ observation of streptavidin-biotin binding on an immunoassay well surface using an atomic force microscope. AB - Polystyrene microtitre wells are commonly used as supports for the enzyme-linked immunosorbent assay (ELISA) method of biomolecular detection, which is employed in the routine diagnosis of a variety of medical conditions. We have used an atomic force microscope (AFM) to directly monitor specific molecular interactions between individual streptavidin and biotin molecules on such wells. This was achieved by functionalising an AFM probe with biotin and monitoring the adhesive forces between the probe and a streptavidin coated immunoassay well. The results demonstrate that the AFM may be employed as an analytical tool to study the interactions between biomolecules involved in immunoassay systems. PMID- 8706851 TI - Up-regulation of cyclooxygenase-2 mRNA in the rat spinal cord following peripheral inflammation. AB - Prostaglandins (PG) have been described as mediators in spinal nociceptive processing after peripheral inflammation. Enzymes essential for PG biosynthesis, cyclooxygenase isozymes COX-1 and COX-2, have not yet been investigated in the spinal cord. In two studies on rats with adjuvant-induced peripheral inflammation levels of mRNA expression of both COX isoforms were analyzed in the lumbar section of the spinal cord using reverse transcription-polymerase chain reaction (RT-PCR) technique. We could show that mRNA of both COX isoforms is expressed constitutively in the spinal cord with COX-2 as the predominant isoform. Six hours after induction of peripheral inflammation, levels of COX-2 mRNA expression were raised significantly in respect to untreated control rats and returned to baseline within 3 days after induction of inflammation. COX-2 might therefore be regarded as the COX isozyme responsible for spinal PG release in nociceptive processing under a peripheral inflammatory stimulus. PMID- 8706852 TI - Growth inhibition by Abl requires an interplay of its SH2 and tyrosine kinase domains. AB - Overexpression of c-Abl tyrosine kinase can be growth inhibitory in certain fibroblast cell lines. Using a series of conditional chimeras between Abl and Src, we have now further dissected the Abl protein to determine which domains are required for this function. We found that growth inhibition, unlike transformation by oncogenic forms of Abl, is dependent on the presence of the cognate SH2 and tyrosine kinase domains. Since growth inhibition correlates with low tyrosine kinase activity, it may involve highly specific interactions of target proteins with both domains without the processivity of phosphorylation associated with oncogenic Abl. PMID- 8706853 TI - An Arabidopsis thaliana cDNA encoding PS II-X, a 4.1 kDa component of photosystem II: a bipartite presequence mediates SecA/delta pH-independent targeting into thylakoids. AB - Higher plant photosystem II preparations contain a 4.1 kDa polypeptide (subunit X) associated with the oxygen-evolving core complex. We describe the isolation of a cDNA encoding PS II-X from Arabidopsis thaliana, in which the C-terminal region is highly homologous to partially sequenced PS II-X from wheat and spinach. The mature protein of 42 residues is preceded by a 74-residue, bipartite presequence similar to those involved in the targeting of nuclear-encoded thylakoid lumen proteins, although hydrophobicity analysis indicates the presence of a single transmembrane span in the mature protein. Moreover, import of pre-PS II-X into the thylakoid membrane of isolated chloroplasts is unaffected by inhibitors of either the Sec- or delta pH-dependent thylakoidal protein translocases, suggesting a spontaneous insertion mechanism. PS II-X appears to be encoded as a mature protein by the plastid genome in the chlorophyll a+c- containing alga, Odontella sinensis. We thus propose that the thylakoid transfer signal of Arabidopsis pre-PS II-X represents a recent acquisition, in phylogenetic terms, compared with signals of Sec-dependent lumenal proteins. PMID- 8706855 TI - Gluconeogenesis from ascorbic acid: ascorbate recycling in isolated murine hepatocytes. AB - Ascorbic acid synthesis and breakdown were investigated in isolated hepatocytes prepared from fasted mice. Stimulation of gluconeogenesis by alanine or xylitol led to ascorbate synthesis. On the other hand, ascorbate or dehydroascorbate addition resulted in concentration-dependent glucose production and elevation of the pentose phosphate pathway intermediate xylulose 5-phosphate. Stimulation of ascorbate oxidation and/or the inhibition of dehydroascorbate reduction increased glucose formation. Inhibition of the pentose phosphate pathway decreased glucose production from dehydroascorbate with increased accumulation of xylulose 5 phosphate. These results suggest that ascorbate can be recycled by a novel way involving intermediates of the pentose phosphate pathway, gluconeogenesis and hexuronic acid pathway. PMID- 8706854 TI - 4-O-phosphoryl-L-threonine, a substrate of the pdxC(serC) gene product involved in vitamin B6 biosynthesis. AB - The Escherichia coli pdxC(serC) gene codes for a transaminase (EC 2.6.1.52). The gene is involved in both pyridoxine (vitamin B6) and serine biosynthesis and was overexpressed as a MalE/PdxC(SerC) fusion protein. The fusion protein was purified by affinity chromatography on an amylose resin and hydrolyzed in the presence of protease factor Xa. Both the fusion protein and the PdxC(SerC) protein were characterized (K(M) value, turnover number, optimum pH). Both enzymes used 4-O-phosphoryl-L-threonine rather than 4-hydroxy-L-threonine as a substrate indicating that the phosphorylated rather than the non-phosphorylated amino acid is involved in pyridoxine biosynthesis. Pyridoxal phosphate was shown to be the cofactor for both enzymes and therefore seems to be involved in its own biosynthesis. PMID- 8706856 TI - Compromised inhibition of human lung lavage cell elastases. AB - Increased elastinolytic activity has been correlated with the degree of lung damage occurring in a variety of lung diseases including cystic fibrosis; serine proteinase inhibitors are currently on trial for the treatment of some lung disorders. However, human lung lavage cells also secrete metallo-dependent elastases. Here we show, for the first time, that whilst these are readily inhibited by EDTA, inhibition of serine elastases using serpins (serine proteinase inhibitors) is not always possible. This may reflect inactivation of serpins by uninhibited metalloproteinases and oxidants in a low protein milieu. Thus, the therapeutic inhibition of excessive elastinolytic activity may require a combination of inhibitors to work efficiently. PMID- 8706857 TI - Regulation of transcription in mammalian cells by yeast Leu3p and externally supplied inducer. AB - The Leu3 protein of yeast is a dual-function regulator, stimulating transcription when the inducer alpha-isopropylmalate (alpha-IPM) is present and suppressing transcription when the inducer is absent. Here we show that Leu3p retains both its positive and negative regulatory properties when expressed in mammalian cells or when added to a mammalian nuclear extract. Alpha-IPM stimulates reporter gene expression 15-20-fold, both in vivo and in vitro. The concentration of alpha-IPM required for half-maximal stimulation in vitro is 2.5 x 10(-4) M. No yeast specific factors other than Leu3p itself are required for up- or down-regulation. Since alpha-IPM is not metabolized in mammalian cells, the Leu3p-alpha-IPM system might be useful in gene therapy and other studies as a highly specific, externally controlled on/off switch of gene expression. PMID- 8706859 TI - Alcoholytic deblocking of N-terminally acetylated peptides and proteins for sequence analysis. AB - N-terminal acetylation of polypeptides is a common feature preventing direct Edman degradation. We describe a method for the removal of the acetyl group, with only a low extent of internal peptide bond cleavage, also in large proteins, by treatment at room temperature with trifluoroacetic acid and methanol. The alcohol is essential for selective deacetylation, and it is proposed that the deblocking mechanism consists of an acid-catalyzed nucleophilic substitution involving methanol. The extent of deacetylation is limited, but the initial yield in the sequence analysis can be up to 10%. Deblocking of samples spotted or blotted onto sequencer filters is equally possible as the use of isolated samples from column separations. Deblocking on sequencer filters is also possible directly after negative results on initial sequencer attempts with samples proving to be blocked. PMID- 8706858 TI - Identification of the functional region on the superantigen Yersinia pseudotuberculosis-derived mitogen responsible for induction of lymphocyte proliferation by using synthetic peptides. AB - Yersinia pseudotuberculosis-derived mitogen (YPM) is the unique Gram-negative bacillary superantigen known. In order to identify the regions on the YPM molecule involved in its superantigenic activity, seven overlapping peptides of the entire YPM molecule were synthesized and tested to evaluate their effects on the YPM-induced proliferation of human peripheral blood lymphocytes. A peptide corresponding to the N-terminal amino acid sequence (1-23) was found to inhibit YPM-induced lymphocyte proliferation in a concentration-dependent manner. The N terminal peptide was found to show no inhibition of the proliferation induced by the other superantigen (staphylococcal enterotoxin B) or the other T-cell mitogen pertussis toxin, indicating that the inhibition is specific to YPM-induced proliferation. Thus, we have identified the N-terminal region (1-23) of the YPM as one of the functional regions responsible for its superantigenic activity. PMID- 8706860 TI - The inhibitory effect of nitrite, a stable product of nitric oxide (NO) formation, on arginase. AB - Macrophages contain arginase and an inducible NO synthase, demonstrated by using L-arginine, the common substrate, for production of both nitric oxide and urea. Arginase was inhibited by nitrite, the stable end product of NO. This inhibition was non-competitive, and could not be explained by the reaction of nitrite with arginine, or by the irreversible covalent modification of arginase, or by the removal of Mn2+, a cofactor of arginase. PMID- 8706861 TI - Identification and characterisation of a homologue of p64 in rat tissues. AB - Previous work has suggested that the gene encoding p64, a component of a bovine kidney intracellular chloride channel, may be a member of a gene family. We have raised a polyclonal antibody to an E. coli fusion protein which has sequence similarity to p64. Immunoblotting detected a protein in rat brain, kidney, liver and lung. In rat brain, the protein was enriched in cerebellar microsomal membranes. Western blot analyses of denaturing and blue native polyacrylamide gels indicated that the protein is a single non-disulphide-linked polypeptide chain with an apparent M(r) of 43 kDa that contributes to a native protein complex with an apparent M(r) of 130 kDa. PMID- 8706863 TI - Activation of G protein-coupled inward rectifier K+ channels in brain neurons requires association of G protein beta gamma subunits with cell membrane. AB - In cultured noradrenergic neurons from the rat locus coeruleus, application of recombinant G protein beta 1 gamma 2 subunits (30 nM) to the cytoplasmic side induced single channel activity similar to the somatostatin-induced single channel activity of G protein-coupled inward rectifier potassium channels (Kir (G)). In contrast, recombinant GTP gamma S-activated, myristoylated alpha i2 (100 nM) did not activate this brain Kir (G). Application of beta 1 gamma 2 C68S (30 nM or 150 nM), in which the cysteine residue fourth from the carboxyl terminus of gamma 2 was replaced by serine, failed to activate the brain Kir(G). This mutant lacks prenylation which is required for the association of beta gamma subunit with the cell membrane. Thus, our results suggest that the association of beta gamma subunit with the cell membrane is a prerequisite for activating Kir(G) channels. PMID- 8706862 TI - A covalently bound catalytic intermediate in Escherichia coli asparaginase: crystal structure of a Thr-89-Val mutant. AB - Escherichia coli asparaginase II catalyzes the hydrolysis of L-asparagine to L aspartate via a threonine-bound acyl-enzyme intermediate. A nearly inactive mutant in which one of the active site threonines, Thr-89, was replaced by valine was constructed, expressed, and crystallized. Its structure, solved at 2.2 A resolution, shows high overall similarity to the wild-type enzyme, but an aspartyl moiety is covalently bound to Thr-12, resembling a reaction intermediate. Kinetic analysis confirms the deacylation deficiency, which is also explained on a structural basis. The previously identified oxyanion hole is described in more detail. PMID- 8706864 TI - Convenient fluorometric assay for matrix metalloproteinase activity and its application in biological media. AB - Matrix metalloproteinases (MMPs) are involved in physiological tissue remodeling and pathological conditions like tumour metastasis and joint destruction. Until now, no convenient and sensitive MMP-activity assay in crude media like synovial fluid has been available. Therefore, the highly soluble fluorogenic substrate TNO211 (Dabcyl-Gaba-Pro-Gln-Gly-Leu-Glu(EDANS)-Ala-Lys-NH2), containing the MMP cleavable Gly-Leu bond and EDANS/Dabcyl as fluorophore/quencer combination, was synthesized and characterized as an MMP specific substrate. We show that the fluorogenic assay using TNO211 is sensitive and can detect MMP activity in culture medium from endothelial cells and untreated synovial fluid (SF) from RA and OA patients, and control subjects. MMP activity in SF significantly increased in the order C < OA < RA, thus the frequent use of OA samples as control in studies on RA is debatable. PMID- 8706865 TI - Probing the higher order structure of RNA with peroxonitrous acid. AB - Potassium peroxonitrite (ONOOK) and [Fe(EDTA)]2- were used to analyze the influence of chemically entirely different hydroxyl radical sources on tRNA cleavage profiles. [Fe(EDTA)]2- gives rise to hydroxyl radicals via a Fenton-like reaction during the oxidation of chelated Fe2+, while ONOOK generates hydroxyl radicals via its conjugate acid (ONOOH) when adding a stable alkaline solution of ONOOK in samples buffered at neutral pH. [Fe(EDTA)]2- is known to induce oxidative strand scission at sugar moieties thought to be solvent accessible, while those residues located in the 'inside' of structured RNAs are protected. Although ONOOH is neutral and significantly smaller than the metal complex, both reagents generate the same protection pattern on tRNAs, suggesting that access of the commonly formed hydroxyl radical, rather than access of its source, is the determining factor when probing the higher order structure of RNA. Strong difference in reactivity is only seen at the modified 2-thiouridine S34 of tRNA(Lys3) which shows hyperreactivity towards ONOOK treatment. This particular reaction may require interaction between the peroxonitrite anion and the thiocarbonyl group of the base, since hyperreactivity is not observed when probing the dethiolated tRNA(Lys3). PMID- 8706866 TI - Fructose 2,6-bisphosphate induces irreversible transitions in cell-free extracts of rat liver. AB - The effect of fructose 2,6-bisphosphate on the dynamics of the 6-phosphofructo-1 kinase/fructose-1,6-bisphosphatase cycle is investigated in a cell-free extract of rat liver under steady-state conditions. Bistability emerges on the basis of the reciprocal allosteric modulation of 6-phosphofructo-1-kinase and fructose-1,6 bisphosphatase. Under conditions of bistability fructose 2,6-bisphosphate may cause transitions between alternative steady states. However, in contrast to what is frequently observed in bistable systems, within a broad range of experimental conditions these transitions proceed irreversibly from states with high ATP to states characterized by low ATP. PMID- 8706867 TI - CPP32 inhibition prevents Fas-induced ceramide generation and apoptosis in human cells. AB - Intracellular activation of sphingomyelinase, leading to ceramide generation, and ICE-like proteases have been implicated in TNF and Fas-induced apoptosis, but the links between these intracellular apoptotic mediators remain undefined. We show here that a specific peptide inhibitor of the ICE-like protease CPP32/Yama (DEVD CHO) blocks anti-Fas-induced apoptosis in Jurkat and U937 cells, while having no effect on TNF-induced apoptosis in U937 cells. This peptide also prevents ceramide accumulation induced by Fas engagement. Jurkat and U937 cells, as well as their mtDNA-depleted derived lines (rho degree cells), were sensitive to ceramide toxicity, which was not prevented by ICE-like protease inhibitors. These results, taken together, suggest that ICE-like protease activation is a prerequisite for ceramide generation and subsequent apoptosis, at least in the case of Fas-induced cell death. PMID- 8706868 TI - Molecular cloning and expression of the human melanocyte stimulating hormone receptor cDNA (FEBS 11553). PMID- 8706869 TI - Macromolecular crowding and the mandatory condensation of DNA in bacteria. AB - Cellular DNA in bacteria is localized into nucleoids enclosed by cytoplasm. The forces which cause condensation of the DNA into nucleoids are poorly understood. We suggest that direct and indirect macromolecular crowding forces from the surrounding cytoplasm are critical factors for nucleoid condensation, and that within a bacterial cell these crowding forces are always present at such high levels that the DNA is maintained in a condensed state. The DNA affected includes not only the preexisting genomic DNA but also DNA that is newly introduced by viral infection, replication or other means. PMID- 8706870 TI - Linguistic analysis of protein folding. AB - Folding of nascent chains resembles the decoding of spoken language in that information is emitted as a unidirectional, one-dimensional string of elements, with higher structures and long-distance interactions emerging with time. Applying a "pseudolinguistic' analysis of structure to a set of all 36 possible six-stranded antiparallel beta-sandwich topologies reveals new order principles and reduces the complexity of this family significantly. The simple connectivity diagrams ("linguistic trees') proposed here allow predictions of the speed and cooperativity of beta-sheet folding and help understanding the cotranslational folding from the N-terminus. PMID- 8706872 TI - Induction of glyoxylate cycle enzymes in rat liver upon food starvation. AB - The key enzymes of the glyoxylate cycle, isocitrate lyase and malate synthase, have been detected in liver of food-starved rats. Activities became measurable 3 days and peaked 5 days after the beginning of starvation. Both enzymes were found in the peroxisomal cell fraction after organelle fractionation by isopycnic centrifugation. Isocitrate lyase was purified 112-fold by ammonium sulfate precipitation, and chromotography on DEAE-cellulose and Toyopearl HW-65. The specific activity of the purified enzyme was 9.0 units per mg protein. The K(m)(isocitrate) was 68 microM and the pH optimum was at pH 7.4. Malate synthase was enriched 4-fold by ammonium sulfate precipitation. The enzyme had a K(m)(acetyl-CoA) of 0.2 microM, a K(m)(glyoxylate) of 3 mM and a pH optimum of 7.6. PMID- 8706871 TI - Heteromeric channel formation and Ca(2+)-free media reduce the toxic effect of the weaver Kir 3.2 allele. AB - Weaver mice have a severe hypoplasia of the cerebellum with an almost complete loss of the midline granule cells. Recent genetic studies of weaver mice have identified a mutation resulting in an amino acid substitution (G156S) in the pore of the inwardly rectifying potassium channel subunit Kir 3.2. When expressed in Xenopus oocytes the weaver mutation alters channel selectivity from a potassium selective to a nonspecific cation-selective pore. In this study we confirm by cell-attached patch-clamp recording that the mutation produces a non-selective cation channel. We also demonstrate that the cell death induced by weaver expression may be prevented by elimination of calcium from the extracellular solution as well as by coexpression with the wild-type Kir 3.2 allele, or other members of the Kir 3.0 subfamily. These results suggest that the weaver defect in Kir 3.2 may cause cerebellar cell death by cell swelling and calcium overload. Cells which express the weaver subunit, but which normally survive, may do so because of heteromeric subunit assembly with wild-type subunits of the Kir 3.0 subfamily. PMID- 8706873 TI - Fungal ferritins: the ferritin from mycelia of Absidia spinosa is a bacterioferritin. AB - Two distinct ferritin like iron containing proteins have been identified and isolated from the fungus Absidia spinosa; one from the spores and another from the mycelia. The mycelial protein has been purified and consists of two subunits of approx. 20 kDa. The N-terminal sequences of both subunits have been determined. The holoprotein as isolated contains approx. 750 iron atoms/molecule and exhibits a heme-like UV-Vis spectrum. Based on the heme spectrum and the high degree of sequence homology found, it has been established that the mycelial protein is a bacterioferritin. This is the first example demonstrating the presence of a bacterioferritin in a eukaryotic organism. PMID- 8706874 TI - Recombinant human proteinase 3, the Wegener's autoantigen, expressed in HMC-1 cells is enzymatically active and recognized by c-ANCA. AB - We developed a stable expression system for conformationally intact recombinant human PR3 (rPR3) using the human mast cell line HMC-1. Like in U937 cells, the rPR3 is processed from a 34 kDa precursor to the 29 kDa mature form, primarily as the result of oligosaccharide trimming. The rPR3 binds [3H]DFP and hydrolyzes the substrate N-methoxysuccinyl-Ala-Ala-Pro-Val-pNA. The enzymatic activity is inhibited by greater than 95% by alpha 1-PI. The rPR3 and the enzymatically inactive mutant rPR3-S176A are both packaged in granules. Thus, proteolytic autoprocessing is not required for PR3's targeting to granules. This rPR3 is the first to be recognized by most c-ANCA from WG patients and all anti-PR3 ANCA that were detected by standard anti-PR3 specific ELISA. This expression system for rPR3 represents a versatile tool for the analysis of its intracellular processing, structure-function relationships and interaction with autoantibodies. PMID- 8706875 TI - Identification of N-acetylglucosamine binding residues in Griffonia simplicifolia lectin II. AB - Primary structure and crystallographic data of several legume lectins were used to predict the involvement in carbohydrate binding of six amino acid residues (Asp88, Glu108, Tyr134, Asn136, Leu226 and Gln227) in Griffonia simplicifolia lectin II (GS-II). The functional involvement of these residues was evaluated by assessing GlcNAc binding of modified forms of GS-II in which these residues were eliminated in truncated peptides or systematically substituted with other amino acids by site-specific mutations. Mutations at Asp88, Tyr134 or Asn136 eliminated GlcNAc binding activity by GS-II, while those at Glu108, Leu226 or Gln227 did not alter the activity. The former three amino acids were functionally essential for carbohydrate binding by GS-II presumably through hydrogen bonding to and hydrophobic interactions with GlcNAc. Although an Asp or Gly substitution for Tyr134 eliminated GlcNAc affinity, substitution with Phe did not appreciably affect binding. Despite the fact that mutations to Leu226 and Gln227 did not alter carbohydrate binding, a truncated form of GS-II lacking these residues no longer exhibited carbohydrate binding affinity. PMID- 8706876 TI - Both action potentials and variation potentials induce proteinase inhibitor gene expression in tomato. AB - Tomato plants (Lycopersicon esculentum) accumulate proteinase inhibitor 2 (pin2) mRNA in response to insect attack, crushing and flaming in leaves distant from those treated. Most earlier work suggests that the systemic wound signals are chemical; here we try to determine whether electrical or physical (hydraulic) signals can also evoke pin expression. We used a mild flame to evoke a systemic hydraulic signal and its local electrical aftermath, the variation potential (VP), and we used an electric stimulus to evoke a systemic electrical signal, the action potential (AP). We determined the kinetic parameters of both the VP and AP. Flame-wounded plants essentially always exhibited major electrical responses throughout the plant and a several-fold increase in pin2 mRNA within 1 h. Electrically stimulated plants that generated and transmitted a signal (AP) into the analyzed leaf exhibited similarly large, rapid increases in pin2 mRNA levels. Plants which generated no signal, or signals of just a few microvolts, had unchanged levels of pin2 mRNA. Since the AP and VP both arrived in the receiving leaf before accumulation of pin2 mRNA began, we conclude that, in addition to the previously shown chemical signals, both hydraulically induced VPs and electrically induced APs are capable of evoking pin2 gene expression. PMID- 8706877 TI - Do voltage-gated Kv1.1 and inward rectifier Kir2.1 potassium channels form heteromultimers? AB - Possible heteromultimer formation between Kv- and Kir-type K+ channels was investigated, in connection with the known functional diversity of K+ channels in vivo. Voltage-clamp experiments were performed on Xenopus oocytes, either injected with concatenated Kir2.1-Kv1.1 mRNA, or co-injected with Kv1.1 and Kir2.1 mRNA. K+ currents could be approximated by the algebraic sum of the 2 K+ current types alone. The tandem construct did not show functional expression, although it could be detected by Western blotting. We conclude that Kv1.1 and Kir2.1 alpha-subunit proteins fail to assemble and do not contribute functional diversity to K+ channels. PMID- 8706878 TI - Depletion of intracellular calcium stores activates an outward potassium current in mast and RBL-1 cells that is correlated with CRAC channel activation. AB - Highly Ca2+ selective Ca2+ channels activated by store depletion have been recently described in several cell types and have been termed CRAC channels (for calcium release-activated calcium). The present study shows that following store depletion in mast and RBL-1 cells, monovalent outward currents could be recorded if the internal solution contained K+ but not Cs+. The activation of the outward K+ current correlated with the activation of ICRAC, in both time and amplitude, suggesting that the K+ current might be carried by CRAC channels. The amplitude of the outward current was increased if external Ca2+ was reduced or replaced by external Ba2+. The outward K+ conductance might have a physiological role in maintaining the driving force for Ca2+ entry during the activation of CRAC channels. PMID- 8706879 TI - Calmodulin inhibits calcium influx current in vascular endothelium. AB - Utilizing the whole-cell configuration of the patch-clamp technique the effect of calmodulin (CaM) on thapsigargin-induced Ca2+ current has been studied. Addition of several concentrations of CaM to the patch pipette induced concentration dependent inhibition of thapsigargin-induced Ca2+ current in bovine aortic endothelial cells. The effect of CaM was Ca2+ dependent and was not observed when the intracellular Ca2+ was buffered to 1 nM with EGTA. CaM produced two major effects on the thapsigargin-induced Ca2+ current. First CaM slow down activation of the current by thapsigargin from a control value of 16 +/- 5 to 31 +/- 6 s with 1 microM CaM in the pipette solution. The second effect of CaM was to reduce the current amplitude in a concentration-dependent manner. The inhibition of Ca2+ current was observed at the peak of the current and at the sustained current level. The reduction of current at the sustained level was observed 15-20 s after onset of the thapsigargin response. The half inhibitory concentration determined from these experiments was 0.1 microM. These results indicate that CaM can modulate thapsigargin-induced Ca2+ current in this endothelium, suggesting a possible role for CaM in the regulation of store-operated Ca2+ influx. PMID- 8706880 TI - A mutation which disrupts the hydrophobic core of the signal peptide of bilirubin UDP-glucuronosyltransferase, an endoplasmic reticulum membrane protein, causes Crigler-Najjar type II. AB - Crigler-Najjar (CN) disease is caused by a deficiency of the hepatic enzyme, bilirubin UDP-glucuronosyltransferase (B-UGT). We have found two CN type II patients, who were homozygous for a leucine to arginine transition at position 15 of B-UGT1. This mutation is expected to disrupt the hydrophobic core of the signal peptide of B-UGT1. Wild type and mutant B-UGT cDNAs were transfected in COS cells. Mutant and wild type mRNA were formed in equal amounts. The mutant protein was expressed with 0.5% efficiency, as compared to wild type. Mutant and wild type mRNAs were translated in vitro. Wild type transferase is processed by microsomes, no processing of the mutant protein was observed. PMID- 8706881 TI - Sequential activation of three distinct ICE-like activities in Fas-ligated Jurkat cells. AB - ICE family proteases have been implicated as important effectors of the apoptotic pathway, perhaps acting hierarchically in a protease cascade. Using cleavage of endogenous protease substrates as probes, three distinct tiers of ICE-like activity were observed after Fas ligation in Jurkat cells. The earliest cleavage detected (30 min) was of fodrin, and produced a 150 kDa fragment. The second phase of cleavage (50 min) involved PARP, U1-70kDa and DNA-PKcs, all substrates of the CPP32-like proteases. Lamin B cleavage was observed during the third cleavage phase (90 min). Distinct inhibition profiles obtained using a panel of peptide-based inhibitors of ICE-like proteases clearly distinguished the three different cleavage phases. These studies provide evidence for a sequence of ICE like proteolytic activity during apoptosis. The early fodrin cleavage, producing a 150 kDa fragment, identifies an ICE-like activity proximal to CPP32 in Fas induced Jurkat cell apoptosis. PMID- 8706882 TI - Two myogenic regulatory factor transcripts exhibit muscle-specific responses to disuse and passive stretch in adult rats. AB - Levels of myogenic regulatory factor (MRF) transcripts are altered in a muscle specific manner in response to hind limb immobilisation of adult male rats, for a 2 day period, in either a lengthened or shortened position which result in passive stretch or disuse atrophy respectively. Myogenin transcript levels were dramatically elevated in the stretched plantaris but not soleus, whereas the MRF4 transcript was significantly elevated in soleus but not plantaris. Levels of myogenin mRNA were unaffected by disuse in either muscle and MRF4 was markedly lower in plantaris in response to disuse. PMID- 8706883 TI - Cross-linking of SsoII restriction endonuclease to cognate and non-cognate DNAs. AB - Specific and non-specific interactions of SsoII restriction endonuclease (R.SsoII) were probed by the method of covalent attachment to modified DNA containing an active monosubstituted pyrophosphate internucleotide bond instead of a phosphodiester one. R.SsoII with six N-terminal His residues was shown to be cross-linked to duplexes with this type of modification, either containing or not the recognition sequence. Competition experiments with covalent attachment of R.SsoII to activated DNAs demonstrated the similar affinity of the enzyme to cognate and non-cognate DNAs in the absence of cofactor, Mg2+ ions. PMID- 8706884 TI - The level of pancreatic PLA2 receptor is closely associated with the proliferative state of rat uterine stromal cells. AB - Rat uterine stromal cells (U(III)) express pancreatic type PLA2 (PLA2-I) receptor and internalize the enzyme bound to receptors. Here, we investigate the proliferating effect and alterations in binding of PLA2-I. There is a dramatic decline in PLA2-I binding in U(III) cells as they progress from a non-confluent proliferating state (40,000 sites/cell) to a confluent state (1300 sites/cell). Intracellular concentration of PLA2-I changed with the alteration in binding, suggesting that regulation in the PLA2 binding capacity may have important implications in growth control mechanisms. PMID- 8706885 TI - Fusion of reconstituted influenza virus envelopes with liposomes mediated by streptavidin/biotin interactions. AB - Reconstituted influenza virus envelopes (virosomes) containing the viral hemagglutinin (HA) represent an efficient fusogenic cellular delivery system. By interaction of HA with its natural receptors, sialylated lipids (gangliosides) or proteins, virosomes bind to cells and, following endocytic uptake, deliver their contents to the cytosol through fusion from within acidic endosomes. Here, we show that binding to sialic acid is not necessary for fusion. In the presence of streptavidin, virosomes containing a biotinylated lipid fused with liposomes lacking sialic acid if these liposomes also had a biotinylated lipid in their membranes. Moreover, fusion characteristics corresponded well with fusion of virosomes with ganglioside-containing liposomes. PMID- 8706886 TI - Sudden depletion of carbon source blocks translation, but not transcription, in the yeast Saccharomyces cerevisiae. AB - Expression of invertase in the yeast Saccharomyces cerevisiae is greatly delayed when derepression occurs in a medium that lacks a usable carbon source. The delay is not a consequence of defects in the transcription of the SUC2 gene but is due to the impossibility of translating the normal levels of mRNA generated under derepressing conditions. The inhibition of translation in the absence of glucose has to be considered when reporter genes such as E. coli lacZ are used to measure transcription in conditions of carbon source starvation. PMID- 8706887 TI - Restoration of phosphorylation capacity to the dormant half of the alpha-subunits of Na+, K(+)-ATPase. AB - Purified kidney Na+, K(+)-ATPase whose alpha-subunit is cleaved by chymotrypsin at Leu266-Ala267, loses ATPase activity but forms the phosphoenzyme intermediate (EP) from ATP. When EP formation was correlated with extent of alpha-cleavage in the course of proteolysis, total EP increased with time before it declined. The magnitude of this rise indicated doubling of the number of phosphorylation sites after cleavage. Together with previous findings, these data establish that half of the alpha-subunits of oligomeric membrane-bound enzyme are dormant and that interaction of the N-terminal domain of alpha-subunit with its phosphorylation domain causes this half-site reactivity. Evidently, disruption of this interaction by proteolysis abolishes overall activity while it opens access to phosphorylation sites of all alpha-subunits. PMID- 8706888 TI - Surface display on staphylococci: a comparative study. AB - Two different host-vector expression systems, designed for cell surface display of heterologous receptors on Staphylococcus xylosus and Staphylococcus carnosus, respectively, were compared for the surface display of four variants of a 101 amino acid region derived from the G glycoprotein of human respiratory syncytial virus (RSV). Surface localization of the different chimeric receptors was evaluated by a colorimetric assay and by fluorescence-activated cell sorting. It was concluded that the S. carnosus system was better both in the ability to translocate inefficiently secreted peptides and in the number of exposed hybrid receptors. The potential use of the described staphylococci as live bacterial vaccine vehicles or alternatives to filamentous phages for surface display of protein libraries is discussed. PMID- 8706889 TI - Synergistic induction of endothelial tissue factor by tumor necrosis factor and vascular endothelial growth factor: functional analysis of the tumor necrosis factor receptors. AB - Tissue factor expression on the surface of endothelial cells can be induced by tumor necrosis factor (TNF) and vascular endothelial growth factor (VEGF) in a synergistic manner. We have investigated the role of the two different TNF receptors for this synergy. Firstly, stimulation of the 60 kDa TNF receptor (TNFR60) by a mutant of TNF specific for TNFR60 induced responses comparable to wild-type TNF. In contrast, stimulation of TNFR80 by a TNFR80-specific TNF mutein did not result in enhancement of tissue factor expression even in the presence of a suboptimal TNFR60 triggering. Secondly, we tested neutralizing TNF receptor antibodies for inhibition of tissue factor synthesis induced by VEGF and TNF. A TNFR60-specific antibody inhibited tissue factor production over a broad range of TNF concentrations, indicating an essential role of TNFR60 in the TNF/VEGF synergy. In contrast, blocking of TNF binding to TNFR80 strongly inhibited TNF induced tissue factor expression at low, but less pronounced at high, TNF concentrations. In conclusion, these data are in agreement with a model in which TNFR80 participates in the synergy between VEGF and low concentrations of soluble TNF by passing the ligand to the signalling TNFR60. PMID- 8706890 TI - The active sites of cellulases are involved in chiral recognition: a comparison of cellobiohydrolase 1 and endoglucanase 1. AB - The cellulases cellobiohydrolase 1 (CBH 1) and endoglucanase 1 (EG 1) from the fungus Trichoderma reesei are closely related with 40% sequence identity and very similar in structure. In CBH 1 the active site is enclosed by long loops and some antiparallel beta-strands forming a 40 A long tunnel, whereas in EG 1 part of those loops are missing so that the enzyme has a more common active site groove. Both enzymes were immobilized on silica and these materials were used as chiral stationary phases for chromatographic separation of the enantiomers of two chiral drugs, propranolol and alprenolol. The CBH 1 phase showed much better resolution than did the EG 1 phase, suggesting that the tunnel structure of the protein may play an important role in the chiral separation. The chiral compounds were found to be competitive inhibitors of both enzymes when p-nitrophenyl lactoside (pNPL) was used as substrate. (S)-enantiomers showed stronger inhibitory effects and also longer retention time on the stationary phases than the (R)-enantiomers. The consistency between kinetic data and retention on the stationary phases clearly shows that the enzymatically active sites of CBH 1 and EG 1 are involved in chiral recognition. PMID- 8706891 TI - Cloning of a novel ubiquitin-conjugating enzyme (E2) gene from the ciliate Paramecium tetraurelia. AB - We isolated a 1.7 kb gene (UbcP1) for a ubiquitin-conjugating enzyme from a P. tetraurelia cDNA library and sequenced it. Its deduced polypeptide sequence consists of 425 amino acid residues (48 kDa). The UbcP1 protein contains novel N- and C-terminal extensions in addition to a UBC domain, and within the UBC domain it shares low identity with sequences of other known E2s. A constructed phylogenetic tree suggests that the UbcP1 protein may represent a member of a distinct subfamily of E2s. Southern blot analysis showed that the N-terminal extension of the UbcP1 is conserved in P. multimicronucleatum. PMID- 8706892 TI - MoaA of Arthrobacter nicotinovorans pAO1 involved in Mo-pterin cofactor synthesis is an Fe-S protein. AB - MoaA, involved in an early step in the biosynthesis of the molybdopterin cofactor (MoCo), has not yet been characterized biochemically and the reaction it catalyzes is unknown. We overexpressed MoaA from pAO1 of Arthrobacter nicotinovorans in Escherichia coli as a N-terminal fusion with either glutathione S-transferase or a 6-histidine tag. The pAO1 encoded MoaA as well as the fusion proteins functionally complement E. coli moaA mutants. Here we show that purified MoaA contains approximately 4 microM Fe and approximately 3 microM acid-labile S/microM protein. EPR spectroscopy revealed a predominant signal at g(av) = 2.01, indicative of a [3Fe-xS] cluster. PMID- 8706893 TI - Co-expression of mRNA for type I and type II interleukin-1 receptors and the IL-1 receptor accessory protein correlates to IL-1 responsiveness. AB - Three cell surface molecules participate in Interleukin-1 (IL-1) binding and signal generation, the two distinct types of receptors (type I IL-1R and type II IL-1R) and the IL-1 receptor accessory protein (IL-1RAcP). Low surface expression hampers the detection of all three components on a protein level in most cell types, thus the highly sensitive RT-PCR was used to analyse the mRNA expression in a panel of 18 murine cell types of different hemopoietic lineages and fibroblasts. The transcription of both types of IL-1 receptors was detected in all cell lines tested. In most cell lines the IL-1RAcP was co-expressed with the IL-1 receptors, and only these lines responded to IL-1. However, in three cell lines no mRNA for the IL-1RAcP could be detected, and these cells did not respond to IL-1. These results suggest that the expression of the IL-1RAcP correlates with IL-1 responsiveness and they point to a pivotal role for the IL-1RAcP in IL 1 signal generation. PMID- 8706894 TI - High-molecular-weight kininogen binds two molecules of cysteine proteinases with different rate constants. AB - Fluorescence titrations showed that high-molecular-weight kininogen binds two molecules of papain, cruzipain and cathepsin S with high affinity. The 2:1 binding stoichiometry was confirmed by stopped-flow kinetic measurements of papain binding, which also revealed that the two sites bind the enzyme with different association rate constants (kass,1 = 23.0 x 10(6) M-1 s-1 and kass,2 = 3.4 x 10(6) M-1 s-1). As for low-molecular-weight kininogen, comparison of these kinetic constants with previous data for intact low- and high-molecular-weight kininogen and the separated domains indicated that the faster-binding site is also the tighter-binding site and is that of domain 3, whereas the slower binding, lower-affinity site is on domain 2. The results further demonstrate that there is no appreciable steric interference between the two domains or by the kininogen light chain in the binding of proteinases. Similarly, the binding of kininogen via its light chain to a surface, as indicated by the binding to the model surface, heparin, did not affect the inhibitory properties of kininogen. The M(r) of high-molecular-weight kininogen was determined to be 83,500 by sedimentation equilibrium measurements, in agreement with the value calculated from amino acid sequence and carbohydrate analysis. PMID- 8706895 TI - Involvement of a single-stranded DNA binding protein, ssCRE-BP/Pur alpha, in morphine dependence. AB - We have purified a nuclear protein from mouse cerebella that binds to single stranded oligo-DNA of cAMP response element and is modulated by morphine treatment. Isolation of the cDNA clone showed that the nuclear protein (ssCRE-BP) was identical to Pur alpha, a DNA binding protein for single-stranded purine-rich sequences that was originally isolated as a replication factor. ssCRE-BP/Pur alpha and mRNA were abundant in the brain. The levels of ssCRE-BP/Pur alpha and the transcript were not changed by chronic morphine treatment, however, the levels of an activator of ssCRE-BP/Pur alpha, which is necessary for the DNA binding, may be modulated by the treatment. PMID- 8706896 TI - Effect of sequence contexts on misincorporation of nucleotides opposite 2 hydroxyadenine. AB - Twelve oligonucleotides containing 2-hydroxyadenine (2-OH-Ade) with different neighboring bases were used as templates in DNA polymerase reactions,and the effects of the sequence contexts were investigated. DNA polymerases alpha and beta inserted dTMP and dCMP opposite 2-OH-Ade in most of the oligonucleotides tested. The Klenow fragment of DNA polymerase I primarily incorporated dTMP and dGMP. Effects of the 5'-flanking base of 2-OH-Ade was found when the 3'-flanking base of 2-OH-Ade was A or C. Incorporation of dAMP occurred when the oxidized base was located in a 5' -TA*A- 3' (A* represents 2-OH-Ade) sequence. These results suggest that the formation of 2-OH-Ade in DNA may induce all the mutations involving A (A-->G transition, and A-->T and A-->C transversions) in cells. PMID- 8706897 TI - Cyclic GMP potentiates phenylephrine but not cyclic ADP-ribose-evoked calcium release from rat lacrimal acinar cells. AB - In the present study, we describe a role for cyclic GMP (cGMP) in the signalling pathway that leads from alpha-adrenergic receptor activation to intracellular Ca2+ mobilization in rat lacrimal acinar cells. The alpha-adrenergic agonist, phenylephrine, stimulates intracellular Ca2+ release which is blocked by inhibitors of guanylate cyclase and cGMP-dependent protein kinase Ia. The membrane-permeable cGMP analogues, dibutyryl-cGMP and 8-bromo-cGMP, potentiate ( approximately 5-fold) the Ca2+ response to submaximal phenylephrine stimulation. In contrast, the same cGMP analogues have no effect on cyclic ADP-ribose-evoked Ca2+ release from permeabilized lacrimal acinar cells. Collectively, these findings suggest that cGMP, via cGMP-dependent protein kinase I alpha , is required for intracellular Ca2+ release following alpha-adrenergic receptor activation in lacrimal acinar cells. PMID- 8706898 TI - In vivo activation of recombinant cAPK catalytic subunit active site mutants by coexpression of the wild-type enzyme, evidence for intermolecular cotranslational phosphorylation. AB - The catalytic subunit of cAMP dependent protein kinase (cAPK) carries two stable autophosphorylated residues. One of them, Thr197, resides in the so-called protein kinase activation segment, and needs to be phosphorylated for full activity and protein kinase inhibitor binding of the enzyme. While wild-type recombinant mammalian C-subunit, expressed in E. coli, can fully autoactivate itself by phosphorylation at Thr197, many active site mutants lack this autophosphorylation activity, so that the primary effects of the mutations become obscured. Two active site mutants of bovine C-subunit, defective in protein kinase inhibitor peptide binding, were activated by wild-type enzyme in vivo, but could not be activated in vitro, demonstrating intermolecular and presumably cotranslational autophosphorylation. The results may delineate strategies for the expression and mutagenesis of other protein kinases with requirements for activation segment phosphorylation. PMID- 8706899 TI - Transport of the glutathione conjugate of ethacrynic acid by the human multidrug resistance protein MRP. AB - The multidrug resistance protein MRP has been shown to mediate the transport of glutathione S-conjugates across membranes. In this study we demonstrate that the glutathione S-conjugate of the diuretic drug ethacrynic acid, which is an efficient inhibitor of glutathione S-transferases, is a high-affinity substrate and inhibitor of the glutathione S-conjugate pump associated with MRP. This implies that ethacrynic acid may modulate drug resistance of tumor cells not only by inhibiting glutathione S-transferase activity, but also by inhibiting the export of drug conjugates from the cell by MRP. PMID- 8706900 TI - Down-regulation of the protein kinase A pathway by activators of protein kinase C and intracellular Ca2+ in fibroblast cells. AB - Many genes are regulated by the intracellular calcium, protein kinase C (PKC) and protein kinase A (PKA) pathways and it has been shown that these pathways synergize in some cell types, whereas they antagonize in others. Here we show that the calcium and PKC pathways suppress the effects mediated by the PKA pathway in a fibroblast cell line. The suppressing effect of elevated intracellular Ca2+ levels, but not of the PKC pathway, can be abrogated by the addition of cyclosporin A (CsA), indicating that the effect of Ca2+ is mediated by phosphatase-2B (PP-2B/calcineurin). Suppression by the PKC pathway is not mediated by the proto-oncogenes c-fos, c-jun and junB, as the co-transfection of these genes does not block the effects of the PKA stimulator 8-Br-cAMP. In addition, cotransfection with the catalytic subunit of PKA shows that the inhibitory effect of PKC occurs upstream of PKA activation. PMID- 8706901 TI - Action of bovine serum albumin on cytochrome c oxidase activity and proton pumping: a role for fatty acids in enzyme function? AB - Bovine serum albumin (BSA) at micromolar concentrations causes a red shift of the Soret band of bovine cytochrome c oxidase with a slow biphasic time course. It also inhibits the turnover of detergent-isolated enzyme in a similarly slow manner; the progress of this inhibition is halted by palmitate and other fatty acids. The inhibitory bovine serum albumin effect may involve fatty acid depletion from the enzyme. Respiration by cytochrome c oxidase vesicles (proteoliposomes) in the presence of ionophores (uncontrolled) shows only a small inhibition by BSA but preincubation of such vesicles with BSA induces a loss of proton pumping activity. After incubation of BSA-depleted proteoliposomes in the presence of reductant with combinations of fatty acids, pumping activity can be fully restored, suggesting a supportive or even essential role of endogenous fatty acids in H+ translocation by this membranous enzyme. PMID- 8706902 TI - Importance of the structure of the RGD-containing loop in the disintegrins echistatin and eristostatin for recognition of alpha IIb beta 3 and alpha v beta 3 integrins. AB - Echistatin and eristostatin are structurally homologous distintegrins which exhibit significant functional differences in interaction with various integrins. We hypothesized that this may reflect differences in the sequences of their RGD loops: 20CKRARGDDMDDYC32 AND 23CRVARGDWNDDYC35, respectively. Mapping of eristostatin peptides obtained by proteolytic digestion suggested that it has the same alignment of S-S bridges as echistatin. Synthetic echistatin D27W resembled eristostatin since it had increased platelet aggregation inhibitory activity, increased potency to block fibrinogen binding to alpha IIb beta 3, and decreased potency to block vitronectin binding to alpha v beta 3 as compared to wild-type echistatin. Since eristostatin and echistatin have a similar pattern of disulfide bridges, we constructed molecular models of eristostatin based on echistatin NMR coordinates. The RGD loops of eristostatin and echistatin D27W were wider than echistatin's due to the placement of tryptophan (rather than aspartic acid) immediately after the RGD sequence. We propose a hypothesis that the width and shape of the RGD loop are important ligand structural features that affect fitting of ligand to the binding pocket of alpha IIb beta 3 and alpha v beta 3. PMID- 8706903 TI - A fungal metabolite mediates degradation of non-phenolic lignin structures and synthetic lignin by laccase. AB - Lignin peroxidase is generally considered to be a primary catalyst for oxidative depolymerization of lignin by white-rot fungi. However, some white-rot fungi lack lignin peroxidase. Instead, many produce laccase, even though the redox potentials of known laccases are too low to directly oxidize the non-phenolic components of lignin. Pycnoporus cinnabarinus is one example of a laccase producing fungus that degrades lignin very efficiently. To overcome the redox potential barrier, P. cinnabarinus produces a metabolite, 3-hydroxyanthranilate that can mediate the oxidation of how non-phenolic substrates by laccase. This is the first description of how laccase might function in a biological system for the complete depolymerization of lignin. PMID- 8706905 TI - Interaction between the left-handed Z-DNA and polyamine-2. The crystal structure of the d(CG)3 and spermidine complex. AB - This paper deals with the crystal structure of d(CG)3-spermidine complex. The DNA fragment, d(CG)3, was crystallized with N-(2-amino-propyl)-1,4-diamino-butane, PA(34), spermidine. The results of its X-ray crystallographic analysis showed many intermolecular contacts between d(CG)3 and spermidine, but the binding mode of spermidine to the d(CG)3 molecule is different from that of the d(CG)3 and N (2-amino-ethyl)-1,4-diamino-butane [PA(24)] complex: a spermidine molecule bound to the d(CG)3 and its symmetrically related neighboring d(CG)3 molecules through the water molecules with hydrogen bonds, while one PA(24) molecule connected directly to one d(CG)3 molecule, but not to its neighboring d(CG)3 molecule. In the crystal, the d(CG)3 molecule was the left-handed Z-form, and three magnesium cations and a sodium cation were observed around the d(CG)3 moiety with different binding modes from the case of the d(CG)3-PA(24) complex. PMID- 8706904 TI - Identification of a novel endogenous memory facilitating cyclic dipeptide cyclo prolylglycine in rat brain. AB - Using high-performance liquid chromatography, gas-chromatography and chromato mass spectrometry methods a novel endogenous cyclic dipeptide cyclo-prolylglycine was identified in rat brain. Its content according to gas chromatography is 2.8 +/- 0.3 nmol/g wet brain. Synthetic cyclo-prolylglycine has demonstrated antiamnesic activity in the passive avoidance test in rats at a dose of 0.1 mg/kg i.p. Cyclic dipeptide cyclo-prolylglycine seems to be a memory facilitating substance and its presence in rat brain suggests the existence of a new mechanism of memory regulation. PMID- 8706906 TI - The three-dimensional structure of capsule-specific CMP: 2-keto-3-deoxy-manno octonic acid synthetase from Escherichia coli. AB - CMP-Kdo synthetases from Gram-negative bacteria activate Kdo for incorporation into lipo- and capsule-polysaccharides. Here we report the crystal structure of the capsule-specific synthetase from E. coli at 2.3 A resolution. The enzyme is a dimer of 2 x 245 amino acid residues assuming C2 symmetry. It contains a central predominantly parallel beta-sheet with surrounding helices. The chain fold is novel; it is remotely related to a double Rossmann fold. A large pocket at the carboxyl terminal ends of the central. beta-strands most likely accommodates the catalytic center. A putative phosphate binding site at the loop between the first beta-strand and the following helix is indicated by a bound iridium hexachloride anion. PMID- 8706907 TI - Structural requirements for alpha-mating factor activity. AB - The sexual hormone of S. cerevisiae, alpha-mating factor (alpha-MF, WHWLQLKPGQPMY) has structural homology with mammalian luteinizing hormone releasing hormone (LHRH, pEHWSYGLRPG-NH2) and has been shown to exhibit LHRH activity [Loumaye et al. (1982) Science 218, 1323-1325]. We have tested whether LHRH has alpha-MF activity in yeast and found that it does not. We therefore synthesized a series of hybrid peptides of alpha-MF and LHRH to study the structural features which determine alpha-MF and LHRH activities. A hybrid peptide consisting of the LHRH sequence with the C-terminal tetrapeptide (QPMY) of alpha-MF did not exhibit alpha-MF activity. Thus, the lack of alpha-MF activity of LHRH is not due solely to the absence of the C-terminal residues. Substitution of Lys7 in alpha-MF with Arg, as is found in LHRH, did not affect the alpha-MF activity, nor did an additional substitution of Trp1 with pGlu. However, the C-terminal four amino acids of alpha-MF were necessary for alpha-MF activity. Our results indicate that insertion of a Ser residue in position 4 as found in LHRH abolishes alpha-MF activity. These results suggest that, in addition to an intact C-terminus, correct spacing of the N-terminal His2 and the C-terminus is required for alpha-MF activity. The hybrid peptides all exhibited less LHRH activity than either LHRH or alpha-MF. These structure-function studies indicate that the structural homology between these two reproductive hormones may not reflect an evolutionary relationship between them. PMID- 8706908 TI - A structural tree for alpha-helical proteins containing alpha-alpha-corners and its application to protein classification. AB - A structural tree for alpha-helical proteins and domains including alpha-alpha corners has been constructed. The alpha-alpha-corner is taken as a root structure of the tree. The larger protein structures are obtained by stepwise addition of alpha-helices to the root alpha-alpha-corner taking into account a restricted set of rules inferred from known principles of protein structure. The protein structures that can be obtained in this way are grouped into one structural class and those found in branches of the tree into subclasses. PMID- 8706909 TI - F protein induced fusion of Sendai viral envelopes with mouse teratocarcinoma cells through Le(x)-Le(x) interaction. AB - The efficiency of membrane fusion between reconstituted Sendai viral envelopes containing only the fusion protein (F-virosomes) and the plasma membrane of mouse teratocarcinoma cells (F9) in culture was assessed using an assay based on the relief of self-quenching of a lipid probe incorporated in the F-virosomes. The potential of F-virosomes was also evaluated for a targeted cytosolic delivery of lysozyme to F9 cells. [125I]Lysozyme entrapped into F-virosomes was taken to examine its fusion-mediated transfer to the F9 cells. Target specificity of the F virosomes was confirmed by the interaction between the terminal Le(x) moiety (Gal beta 1-->4(Fuc alpha 1-->3)GlcNAc) of F protein and the Le(x) determinant on the membrane of F9 cells. Incubation of the loaded F-virosomes with cells led to fusion-mediated delivery, as inferred from the ability of cells to internalize lysozyme in the presence of azide (a potent inhibitor of endocytosis). These results suggest that carbohydrate-carbohydrate interaction is strong enough for target cell recognition followed by phospholipid bilayer melding induced by fusion glycoprotein of Sendai virus. PMID- 8706910 TI - Effect of analogues of diaminopimelic acid on the meso-diaminopimelate-adding enzyme from Escherichia coli. AB - Several analogues of diaminopimelic acid (A2pm) were tested as substrates or inhibitors of the meso-diaminopimelate-adding enzyme from Escherichia coli. They included lanthionine derivatives, a phosphonic analogue, heterocyclic compounds, 3-fluoro-A2pm, 4-methylene-A2pm and N-hydroxy-A2pm. The best substrates were, in decreasing order of specific enzyme activity, (2S,3R,6S)-3-fluoro-A2pm, meso lanthionine sulfoxide and N-hydroxy-A2pm (mixture of stereoisomers). In those cases where all the stereoisomers were available, the specificity could be described as meso > > DD approximately to LL. N-Hydroxy-A2pm (mixture of stereoisomers) strongly inhibited the addition of radioactive meso-A2pm to UDP-N acetylmuramoyl-dipeptide. PMID- 8706911 TI - Levels of endogenous cytokinins, indole-3-acetic acid and abscisic acid during the cell cycle of synchronized tobacco BY-2 cells. AB - Correlation between cell cycle progression and endogenous levels of plant hormones was studied in synchronized tobacco BY-2 cell suspension cultures. Sixteen different cytokinins, indole-3-acetic acid (IAA) and abscisic acid (ABA) were extracted using solid-phase anion exchange chromatography in combination with immunoaffinity purification, and quantified by mass spectrometry. No significant correlation could be identified for IAA and ABA. In contrast, there were sharp peaks in the levels of specific cytokinins (zeatin- and dihydrozeatin type) at the end of the S phase and during mitosis. The levels of other cytokinins analyzed, including zeatins N- and O-glucosides, remained low, suggesting that the increased amounts of their corresponding non-glucosylated form resulted from de novo synthesis. These findings suggest that zeatin- and dihydrozeatin-type cytokinins might play a specific regulatory role in the progression of the plant cell cycle. One hypothesis to explain cytokinin action is based on a specific interaction with kinases that regulate cell cycle progression, as has been recently shown for the cytokinin analogue olomoucine. PMID- 8706913 TI - Heat shock protein hsp70 overexpression confers resistance against nitric oxide. AB - Heat stress is known to render rat islet cells resistant against the toxic effects of nitric oxide, reactive oxygen intermediates and the islet cell toxin streptozotocin. We report here for the first time that protection against nitric oxide is mediated by the major heat shock protein, hsp70, even in the absence of heat stress. The human hsp70 gene was stably transfected into the rat insulinoma cell line RINm5F. Constitutive expression of hsp70 caused protection from NO induced cell lysis which was of the same extent as seen after heat stressing cells. Our results identify hsp70 as a defence molecule against nitric oxide. PMID- 8706912 TI - Neopterin activates transcription factor nuclear factor-kappa B in vascular smooth muscle cells. AB - We have previously shown that the pteridine compound neopterin stimulates inducible nitric oxide synthase (iNOS) gene expression in vascular smooth muscle cells in vitro. The mechanisms whereby neopterin exhibits these effects remained unclear. The present study demonstrates that neopterin induces the translocation of the transcription factor nuclear factor-kappa B (NF-kappa B) to the nucleus. Pretreatment of cells with the antioxidant pyrrolidine dithiocarbamate completely suppressed the effects of neopterin on NF-kappa B activation, iNOS gene expression, and nitric oxide release. From these data we conclude that neopterin activates the translocation of NF-kappa B subunits to the nucleus by modulating the intracellular redox state. This is one possible explanation for the impact of neopterin on iNOS gene expression. PMID- 8706914 TI - Up-regulation of L- and non-L, non-N-type Ca2+ channels by basal and stimulated protein kinase C activation in insulin-secreting RINm5F cells. AB - We studied the effect of protein kinase C (PKC) inhibition and activation on voltage-dependent Ca2+ channels in rat insulinoma RINm5F cells. PKC down regulation by chronic (24 h) treatment with the PKC activator phorbol 12 myristate 13-acetate (PMA) reduced by about 60% the Ba2+ currents through L- and non-L, non-N-type high-voltage-activated Ca2+ channels, indicating that PKC tonically up-regulates the two main Ca2+ channel subtypes of RINm5F cells under basal conditions. Consistently, PKC activation by acute PMA application caused only a modest increase (average 23%) of Ba2+ currents in a minority of cells (24%). L- and non-L, non-N-type channels were differentially up-regulated by either basal or stimulated PKC activation. Acute up-regulation was predominant on L-type channels and caused an I/V shift of the Ba2+ currents in the hyperpolarizing direction. Non-L, non-N-type channels were less affected by acute PMA application, possibly reflecting a more effective tonic PKC up-regulatory action. Unexpectedly, the increase of Ba2+ currents during acute PMA application was followed by a progressive current decrease, which was also observed in isolation in another 24% of the cells and could be ascribed to PKC-induced ATP depletion, rather than to a direct effect of PKC on Ca2+ channels. We also provide evidence that PKC-mediated phosphorylation is not involved in the G protein-mediated noradrenergic modulation of Ca2+ channels in RINm5F cells. PMID- 8706915 TI - Presence of epsilon-adenosine tetraphosphate in chromaffin granules after transport of epsilon-ATP. AB - Adenosine 5'-tetraphosphate (Ap4) is a natural constituent of chromaffin granules with concentration values of 2.2 +/- 0.1 nmol/mg of protein and a ratio 245 +/- 40 times lower with respect to ATP (n = 4). The granular transport of epsilon-ATP resulted in a time- and concentration-dependent production of epsilon-adenosine tetraphosphate (epsilon-Ap4) at the intragranular level. The epsilon-Ap4 formation followed a hyperbolic saturation kinetic at low epsilon-ATP concentrations with K(m) value of 0.4 microM epsilon-ATP intragranular (1.15 pmol/mg of granular protein). Intragranular concentrations of epsilon-ATP higher than 500 pmol/mg of protein (approximately to 175 microM intragranular) resulted in a non-saturable production of epsilon-Ap4. PMID- 8706916 TI - Dependence of critical micelle concentration of a zwitterionic detergent on ionic strength: implications in receptor solubilization. AB - The zwitterionic detergent, 3-[(3-cholamidopropyl)-dimethylammonio]-1 propanesulfonate (CHAPS), is mild, non-denaturing, and extensively used for solubilizing membrane proteins and receptors. We report here that the critical micelle concentration (CMC) of CHAPS is dependent on the concentration of NaCl in the solution. Thus, the CMC of CHAPS decreases from 6.41 mM in absence of any salt to 4.10 mM in presence of 1.5 M NaCl. The logarithm of the CMC values appear to have a linear relationship with the salt concentration. Such changes in CMC with ionic strength have important implications in solubilization of membrane bound neuronal receptors. This is shown by optimal solubilization of the serotonin receptor type 1A (5-HT1A) from bovine brain hippocampal crude (native) membrane by CHAPS at premicellar concentration under high salt conditions. PMID- 8706917 TI - NMR structures of a mitochondrial transit peptide from the green alga Chlamydomonas reinhardtii. AB - The 26-amino-acid pre-sequence of the ATP synthase beta subunit that directs the protein from the cytosol to mitochondria in the unicellular green alga Chlamydomonas reinhardtii has been synthesised and analysed using NMR spectroscopy/circular dichroism and compared to a chloroplast transit peptide from the same organism. The results demonstrate that the peptide, though mainly unstructured in water, undergoes a strong conformational change in a 36% water/64% 2,2,2-trifluoroethanol mixture. In this solvent condition, an alpha helix was characterised by NMR from residue 2 to 26. Structure calculations under NMR restraints lead to a population of models of which 60% are kinked at position 9-10. Structural analysis indicates two hydrophobic sectors on the models with a discontinuity at the 9-10 kink level. The structures suggest a different interaction mode with the mitochondrial membrane compared to the chloroplast transit peptide. PMID- 8706918 TI - Localisation of an ATP-binding site on adenylyl cyclase type I after chemical and enzymatic fragmentation. AB - Photolabeling of partially purified bovine brain adenylyl cyclase (AC I) with [gamma 32P]8-N3-ATP led to incorporation of 32P into the 115 kDa catalyst. Further treatment with N-chlorosuccinimide, which cleaves proteins at tryptophan residues, yielded a 14 kDa 32P-labeled fragment. The latter was immunoprecipitated by antibody BBC1, recognizing the extreme C-terminus of AC I, but not by antibody BBC2, recognizing a more remote epitope. Further fragmentation of photolabeled AC I by the proteases Glu-C and Asp-N yielded 32P labeled peptides corresponding to 2.9 kDa and 5.6 kDa fragments, which were not recognized by any of these antibodies. This narrows the ATP binding site down to a 25 amino acid sequence containing a general motif G(X0-7)KG(X0-4)L/M(X5-7)S/T present in all eukaryotic adenylyl cyclases so far cloned, but also in a variety of bacterial adenylyl cyclases (Peterkofsky et al. (1993) Progr. Nucleic Acids Res. Mol. Biol. 44, 31-65). PMID- 8706919 TI - Nitric oxide donor induces HSP70 accumulation in the heart and in cultured cells. AB - As our group has shown, the NO-synthase inhibitor L-NNA decreased 2-3 times heat shock-induced synthesis of the heat shock protein HSP70 (FEBS Lett. 370 (1995) 159-162). It was suggested that NO is involved in such induction. In the present study, it was found that (1) injection of the NO donor dinitrosyl iron complex (DNIC) into rats results in accumulation of HSP70 in the heart; (2) heat shock is accompanied by increased generation of NO (EPR assay) and HSP70 accumulation in cultured cells; (3) DNIC induces HSP70 accumulation in cultured cells not exposed to heat shock. PMID- 8706920 TI - Conformational change in DNA induced by cationic bilayer membranes. AB - The effect of synthetic cationic lipids on the structure of DNA was studied. The fluorescence enhancement of ethidium bromide on intercalation into DNA was suppressed by the addition of bilayer-forming lipids, but not by micellar ones. Results on the fluorescence depolarization index suggest that ethidium bromide is not released from DNA by lipids intercalated into DNA. CD spectra of the DNA lipid complexes revealed that the structure of DNA was changed only by bilayer forming lipids at temperatures lower than their Tc values. Thus, the conformation of DNA is forced to change by cationic lipids forming the rigid bilayer membrane so that ethidium bromide fluorescence might be reduced, and the conformation can be controlled by selection of the appropriate lipid and temperature. PMID- 8706921 TI - Activation of platelet-derived growth factor (PDGF) receptor dephosphorylation in intact Swiss 3T3 cells by elevators of intracellular Ca2+ and cAMP. AB - We investigated the modulation of platelet-derived growth factor (PDGF) receptor dephosphorylation in Swiss 3T3 cells using a novel assay permitting monitoring of receptor dephosphorylation in intact cells. PDGF treatment of the cells reduced the receptor dephosphorylation rate to 41%, the elevators of intracellular Ca2+, A23187 and thapsigargin increasing it to 227 and 138%, respectively. The cAMP elevators forskolin and isobutylmethylxanthine also accelerated PDGF receptor dephosphorylation. The involvement of Ca(2+)- and cAMP-dependent protein kinases in the regulation of PDGF receptor dephosphorylation is suggested. PMID- 8706922 TI - The role of the T-loop of the signal transducing protein PII from Escherichia coli. AB - The 3D structure of PII, the central protein that controls the level of transcription and the enzymatic activity of glutamine synthetase in enteric bacteria revealed that residues 37-55 form the "T' loop, part of which protrudes from the core of the protein. Within this loop are the only two tyrosine residues that occur in the polypeptide, and one of them, Tyr-51, has been shown by chemical modification studies to be the site of uridylylation. Since tyrosine at position 46 is conserved in all known PII proteins, oligonucleotide directed mutagenesis was used to investigate the role of the two residues. Changing Tyr-51 to phenylalanine or serine abolished uridylylation. Altering tyrosine at position 46 to phenylalanine affected the rate of uridylylation of the protein. This latter mutation does not alter the structure of PII but the reduction in the uridylylation efficiency suggests a role for this residue in recognition and binding of the sensor enzyme uridylyl transferase. PMID- 8706923 TI - The HMG box of SRY is a calmodulin binding domain. AB - The HMG box domain of the testis determining factor, SRY, includes a basic amphiphilic sequence common to calmodulin (CaM) binding proteins. By affinity chromatography, native gel electrophoresis and fluorescence spectroscopy, we show the calcium-dependent binding of SRY to CaM. Binding occurs via the HMG box and an SRY peptide of residues 57-80 binds CaM like the intact domain. SRY/CaM complex formation is specifically inhibited by the SRY DNA binding site sequence, AACAAT, but not a mutated sequence. Fluorescence spectra of the SRY/CaM complex indicate 1:1 stoichiometry and that binding is accompanied by a conformational change in SRY. The A domain of HMG1 also binds CaM and we propose that CaM binding is a property of the wider HMG box family, including SOX and TCF/LEF proteins. These results suggest that CaM may regulate the DNA binding activity of HMG box transcription factors. PMID- 8706924 TI - Unassembled subunits of the photosynthetic oxygen-evolving complex present in the thylakoid lumen are long-lived and assembly-competent. AB - Physiologically healthy pea chloroplasts contain unassembled, soluble subunits of the oxygen-evolving complex in the thylakoid lumen. We report that the lifetimes of two of these subunits, both on and off the membrane, exceed 8 h in vitro. We also demonstrate that each of the subunits present in the thylakoid lumen is competent for assembly into the membrane-bound complex. These data are consistent with the postulate that the soluble lumen-resident subunits play a role in photosystem II homeostasis. We also demonstrate that the reconstitution of the 33 kDa subunit is inhibited by extremely low concentrations of Triton X-100, suggesting that hydrophobic interactions are involved in the binding of this subunit to the photosystem II reaction center. PMID- 8706925 TI - Differences in DNA-binding efficiency of Sp1 to aldolase and pyruvate kinase promoter correlate with altered redox states in resting and proliferating rat thymocytes. AB - Thymocytes induce their glycolytic enzymes as they undergo transition from the resting to the proliferating state. Corresponding increases in mRNA levels point to a transcriptional regulation. Electrophoretic mobility shift assays revealed that the DNA-binding efficiency of Sp1 is increased when nuclear extracts from proliferating compared to resting rat thymocytes were used. Here we demonstrate that hydrogen peroxide, added to nuclear extract from proliferating cells, decreases the Sp1 DNA-binding activity, whereas in nuclear extracts from resting cells dithioerythritol fully restores DNA-binding efficiency. Moreover we show that in contrast to resting thymocytes, production of reactive peroxide anions upon priming with phorbol 12-myristate 13-acetate is nearly abolished in the proliferating cells. From these results we propose that reactive oxygen intermediates affect the interaction of the Sp1 transcription factor with its consensus sequence and subsequently regulate glycolytic gene expression. PMID- 8706926 TI - Regulation of myosin heavy chain expression in adult rat hindlimb muscles during short-term paralysis: comparison of denervation and tetrodotoxin-induced neural inactivation. AB - The extent to which myosin profiles within adult fast and slow muscles are altered by short-term paralysis remains equivocal. We used an array of specific antibodies to identify adult and developmental MHC isoforms within EDL and soleus muscle fibers, and show a marked multiple expression of MHCs with a general shift towards slower and more energy efficient MHC profiles after 2 weeks of denervation or TTX nerve conduction block. Paralysis also induced marked expression of an embryonic MHC within most EDL cell types, and a subtle, paralysis-sensitive, expression of alpha-cardiac MHC within specific EDL and soleus extrafusal fibers. Comparison of treatment groups also permitted assessment of the relative influence of neural activity versus trophic factors on these isoforms, and confirmed activity as a major, but not sole, regulator of MHC expression. PMID- 8706927 TI - Immunohistochemical localisation of the serotonin 5-HT2B receptor in mouse gut, cardiovascular system, and brain. AB - We recently reported the cloning of a new member of the serotonin 5-HT2 family, the 5-HT2B receptor. We now report the production and characterisation of a specific antiserum directed against the C-terminal portion of the mouse 5-HT2B receptor. After affinity purification, this polyclonal antibody recognises specifically the mouse 5-HT2B receptor. Immunohistochemical analysis of cryosections from various adult mouse tissues reveals a major 5-HT2B receptor expression in stomach, intestine and pulmonary smooth muscles as well as in myocardium. Furthermore, the antiserum recognises specific areas of the mouse brain, including cerebellar Purkinje cells and their projection areas. PMID- 8706928 TI - A novel murine cathelin-like protein expressed in bone marrow. AB - A novel cDNA encoding a putative secreted protein was isolated from murine bone marrow. The encoded protein named MCLP (murine cathelin-like protein) was found to be highly homologous to the pig cathelin, and to four neutrophil antimicrobial polypeptides: CAP 18, indolicidin, Bac 5 and FALL-39. Secondary structure prediction studies identified a highly cationic region in the C-terminal part of prepro-MCLP with a tendency to adopt an amphipathic alpha-helical conformation, as observed in many antimicrobial peptides. However, no antibacterial activity was observed with the synthetic peptide corresponding to this region of MCLP. PMID- 8706929 TI - Immunopurification and characterization of a collagenase/gelatinase domain issued from basement membrane fibronectin. AB - The proteolytic potential of cellular fibronectin fragments issued from a basement membrane hydrolysate was investigated. Three different gelatinase activities (47, 43 and 37 kDa), located by gelatin zymography, were isolated using successively heparin-agarose, gelatin-agarose and immunopurification with polyclonal antibodies directed against bovine plasma fibronectin. These fragments were also characterized using a monoclonal antibody directed against the extra domain EDA of cellular fibronectin as a probe. A collagenase activity, reliably indicated by the gelatin zymography pattern, was also found using MCA-Pro-Leu-Gly Leu-DPA-Ala-Arg-NH2, the intramolecularly quenched fluorogenic substrate of collagenases. From these results, cellular fibronectin was found to be able to exhibit a proteolytic function after limited proteolysis. This MMP-like function could be associated with tissue remodeling in both normal and pathological states, such as metastasis, angiogenesis and tissue repair. PMID- 8706930 TI - Dexamethasone enhanced by insulin, but not by thyroid hormones stimulates apolipoprotein B mRNA editing in cultured rat hepatocytes depending on the developmental stage. AB - The increase of hepatic apolipoprotein B (apoB) mRNA editing during rat development was not affected by hypothyroidism. Furthermore, the addition of 3,3',5'-triiodothyronine (T3) to cultured hepatocytes taken from fetal, neonatal and adult rats had no effect on apoB mRNA editing. In contrast, dexamethasone markedly stimulated apoB mRNA editing in hepatocytes taken from neonates. This effect was enhanced by the addition of insulin. For the first time our data provide evidence that glucocorticoids together with insulin are important for the regulation of apoB mRNA editing during postnatal development, whereas thyroid hormones are not critical for this process. PMID- 8706931 TI - Primary structure of a novel ABC transporter with a chromosomal localization on the band encoding the multidrug resistance-associated protein. AB - Complementary DNA clones encoding a novel protein, ABC-C, with the typical structural features of the ABC transporter family were identified in a human medullary thyroid carcinoma cell line. The transporter consists of 1704 amino acid residues with two homologous repeats, each harboring six putative transmembrane helices and an ATP-binding cassette motif. The mRNA is expressed highest in normal lung, but also in varying amounts in other tissues and in C cell carcinoma. The ABC-C gene is mapped on chromosome 16p13.3, in close physical proximity to another ABC transporter, the multidrug resistance-associated protein. This related protein is assumed to confer resistance to chemotherapeutic drugs in small cell lung carcinoma. The genomic clustering of both transporters, typical also for other members of the ABC family, supports the notion that ABC-C may be involved in development of resistance to xenobiotics. PMID- 8706932 TI - Interaction studies between the p21Cip1/Waf1 cyclin-dependent kinase inhibitor and proliferating cell nuclear antigen (PCNA) by surface plasmon resonance. AB - The cyclin-dependent kinase (CDK) inhibitor p21Cip1 consists of two domains that interact with CDKs and proliferating cell nuclear antigen (PCNA), respectively. We have investigated the interaction between p21Cip1 and PCNA using surface plasmon resonance (SPR) technology and compared the results with those obtained from other sources such as the yeast two-hybrid system. Whilst other methods are only semi-quantitative, the SPR technique allowed us to determine the kinetic parameters of the interaction. The apparent equilibrium constant KD calculated for these kinetic parameters was 3.2 x 10(-7) M. We further demonstrate the use of SPR to study the interaction between mutant proteins and to determine their actual KD. The interaction between p21Cip1/PCNA is shown to be dependent upon the trimeric conformation of PCNA since a point mutant that abolishes PCNA-PCNA interaction also abolishes PCNA's interaction with p21Cip1. Finally, we demonstrate that SPR can be used to characterise the interaction of p21Cip1 and PCNA in the presence of short competitive peptides. PMID- 8706933 TI - A model phage display subtraction method with potential for analysis of differential gene expression. AB - In order to establish a subtractive procedure that makes it possible to enrich selectively phage displayed antibodies directed against proteins constituting a difference between two populations of cells, a competitive selection strategy utilising two solid phases was developed and tested. Antibodies recognising a defined difference between two otherwise identical protein mixtures were isolated and their specificity confirmed. To test further the efficacy of selection inhibition during the competitive selections, selections towards a total cell extract were performed with and without competition from the same extract. An analysis of the resulting phage antibodies confirmed the subtractive nature of the system described. PMID- 8706934 TI - Impaired glutathione biosynthesis in the ischemic-reperfused rabbit myocardium. AB - Non-protein thiols (NP-SH) and the activities of the glutathione status regulating enzymes gamma-glutamylcysteine synthetase (G-GCS), gamma-glutamyl transpeptidase (G-GT) and glutathione reductase (GR) were assessed in perfused rabbit hearts subjected to severe (60 min) or mild (7 min) total ischemia and 30 min reperfusion. Severe ischemia significantly decreased NP-SH, which were further depressed on reperfusion together with a significant decline in G-GCS activity; G-GT and GR activities were unchanged. Specific analytes were unaffected by mild ischemia-reperfusion. Thus, impaired enzymatic biosynthesis of GSH is operative in the reperfused rabbit myocardium after 60 min ischemia. This phenomenon may favour myocardial GSH depression and oxidative reperfusion injury after severe ischemia. PMID- 8706935 TI - Basic helix-loop-helix protein MyoD displays modest DNA binding specificity. AB - The expression of MyoD can activate muscle specific genes and myogenic differentiation in many cell types. The hypothesis that the DNA binding specificity of MyoD is responsible for its biological specificity was tested. Homodimers of MyoD bind to E-box containing DNA with high affinity, but do not form stable and well defined complexes with heterologous DNA sequences. The physiologically active heterodimer of MyoD and E12 binds an oligonucleotide containing an E-box sequence with an affinity only two orders of magnitude higher than a completely unrelated DNA sequence, stressing the importance of cooperative interactions with other proteins of the transcriptional machinery for specific gene activation. PMID- 8706936 TI - Stimulation of platelet glycoprotein IIb-IIIa (alpha IIb beta 3-integrin) functional activity by a monoclonal antibody to the N-terminal region of glycoprotein IIIa. AB - Platelet glycoprotein (GP) IIb-IIIa complex (alpha IIb beta 3-integrin) changes its conformation upon platelet activation that results in binding of RGD containing ligands and expression of ligand-induced binding site (LIBS) neoepitopes. Anti-GIIb-IIIa monoclonal antibody (monAB) CRC54 bound to < or = 10% of GPIIb-IIIa on resting platelets but binding was enhanced by the occupation of GPIIb-IIIa with RGDS peptide and by platelet activation indicating that CRC54 is directed against LIBS epitope. The epitope was located within the first 100 N terminal residues of GPIIIa and differed from other LIBS epitopes. CRC54 as well as its Fab fragments were able to induce platelet aggregation. CRC54 also stimulated interaction of GPIIb-IIIa with its ligands (fibrinogen and fibronectin) and conformation-dependent antibodies. The results indicated that changes of GPIIb-IIIa conformation, binding of ligands and platelet aggregation could be stimulated via interaction of anti-LIBS antibody with the N-terminal part of GPIIIa. PMID- 8706937 TI - Differential post-transcriptional control of ornithine decarboxylase and spermidine-spermine N1-acetyltransferase by polyamines. AB - Ornithine decarboxylase (ODC) and spermidine/ spermine N1-acetyltransferase (SSAT) are short-lived polyamine enzymes with rate-limiting roles in controlling polyamine biosynthesis and catabolism, respectively. We have found that treatment of MALME-3M human melanoma cells for 6 h with 10 micrograms/ml cycloheximide (CHX) increases ODC and SSAT mRNA 6-9-fold. When cells containing CHX-induced SSAT mRNA were washed and post-incubated for an additional 6 h in drug free media, enzyme activity increased only 2-fold above that in untreated cells despite the > 6-fold increase in accumulated mRNA. Inclusion of 10 microM spermine or spermidine in the post-incubation medium increased SSAT activity approximately 7-fold without further elevating SSAT mRNA levels. This indicates post-transcriptional regulation which, due to the similarity between polyamine mediated increases in SSAT activity and available mRNA, probably occurs at the level of mRNA translation. In contrast to the SSAT response, polyamines markedly reduced ODC activity (but not mRNA) to one sixth that in cells not exposed to polyamines. The findings illustrate how via post-transcriptional mechanisms, shifts in intracellular polyamine pools can simultaneously and differentially regulate polyamine biosynthesis and catabolism. It is hypothesized that these post-transcriptional responses enable cells to rapidly and sensitively control intracellular spermidine and spermine pools. PMID- 8706938 TI - Crystallization and preliminary X-ray analysis of human brain hexokinase. AB - Human brain hexokinase type I, expressed in Escherichia coli, has been crystallized from polyethylene glycol 8000 in the presence of inorganic phosphate. The crystals are hexagonal needles of diameter 0.25 mm, diffracting to a resolution of 3.5 A on a rotating-anode/area-detector system. The crystals belong to the space group P3(1)21/P3(2)21 with cell dimensions a = b = 171.5 A and c = 99.4 A. The specific volume of the crystal is 4.2 A3/Da, suggesting an asymmetric unit with a single 100 kDa molecule and a solvent content of 71% by volume or two molecules of hexokinase with a solvent content of 41%. The complex of hexokinase with glucose crystallizes under similar conditions, giving crystals of the same morphology. PMID- 8706939 TI - Cell type specific regulation of COUP-TF II promoter activity. AB - COUP-TF family orphan receptors regulate activity of ligand-activated nuclear hormone receptors or function independently in the regulation of gene expression. COUP-TF II has a complex expression pattern suggesting that different mechanisms are involved in the regulation of its expression. We isolated the 5' regulatory region of the mouse COUP-TF II gene and demonstrated that the basal promoter is localized in a -200 bp region 5' from the transcription start site. All-trans retinoic acid and dibutyryl cyclic AMP have cell type specific effects on COUP-TF II promoter activity. The effect of cyclic AMP is mediated by the cyclic AMP response element that is localized 74 nucleotides upstream from the major transcriptional start. In vitro promoter analyses also demonstrated that the effect of all-trans RA is not directly mediated by the binding of RARs or RXRs to the promoter sequence. PMID- 8706940 TI - Expansive benign lesions: treatment by laser. AB - Submucous fibroids, polyps and dysfunctional bleeding are responsible for meno metrorrhagia in women over 40 years of age. The authors describe the technique of hysteroscopic myomectomy and endometrial ablation using the Nd-YAG laser and the advantages of the preoperative use of GnRH agonist therapy. They report their results in a series of 746 patients treated for meno-metrorrhagia (366 cases of myomectomy and 380 cases of endometrial ablation). PMID- 8706941 TI - Etiology of and therapeutic approach to synechia uteri. AB - The universal incidence of synechia uteri (Asherman's syndrome) is steadily increasing. The main offender in the etiology of this disorder is trauma to a pregnant uterus, especially after curettage in puerperium or after missed abortion, whereas the role of infection in traumatic intrauterine adhesions should be viewed with caution. Genital tuberculosis is one of the main etiological factors of this condition. The syndrome is expressed by infertility (43%) and menstrual disorders (62%). Pregnancy, when achieved, may be complicated by premature labor, placenta previa and placenta accreta. The diagnosis is made by hystersalpingography, and mainly by hysteroscopy. Preferred treatment is lysis of adhesions by hysteroscopy, followed by immediate insertion of an intrauterine device, combined with a course of estrogens. The success of treatment regarding term deliveries and rate of abortions depends on the severity of the adhesions. We carried out studies regarding regeneration of the endometrium, and intended to induce intrauterine adhesions as a method of treatment in cases of severe uterine bleeding and as a method of contraception. PMID- 8706942 TI - Prenatal diagnosis by chorionic villus sampling. AB - Chorionic villus sampling (CVS) retains its great advantage over mid-trimester amniocentesis by producing early results. Moreover, rapid analytical techniques reduce significantly the waiting time between sampling and diagnosis, while recombinant DNA technology and human gene mapping progress amplify enormously the spectrum of the indications. The recent inclusion in the prenatal diagnosis package of screening tests based on DNA analysis for the major genetic diseases (i.e. cystic fibrosis, fragile-X mental retardation syndrome) may efficiently contribute to prevent the genetic disease. The role of CVS in twin pregnancy has been investigated and compared to amniocentesis. Although these techniques are equally safe, CVS should be considered the approach of choice for a number of technical advantage and in relation to selective fetal reduction in discordant twins. Recent reports have substantially contributed on the hypothetical relationship between limb reduction defects (LRDs) and chorion biopsy. The analysis of LRDs among more than 130,000 CVS reported to WHO CVS-Registry has been unable to find out any relationship between sampling and fetal malformations, including LRDs. In conclusion, first trimester CVS should be considered the gold standard procedure for prenatal diagnosis of genetic diseases. PMID- 8706943 TI - The combined use of prostaglandin and antiprogestin in human fertility control. AB - This paper is a short review of the combined use of RU486 and prostaglandins in human fertility control. It principally insists on the several prostaglandins which are currently used. PMID- 8706944 TI - Obstetric patients treated in intensive care units and maternal mortality. Regional Teams for the Survey. AB - OBJECTIVE: To ascertain the frequency of serious diseases in pregnant women. STUDY DESIGN: A population based survey was performed in France. The cases were all the women admitted for treatment in intensive care unit (ICU). The severity of the cases was measured with the simplified acute physiology score (SAPS) the lethality and the rate of still birth. RESULTS: 435 obstetric patients were included. The estimated frequency of severe diseases was 310 S.D.36 per 100,000 live births. The most frequent diagnose that motived admission in ICU was hypertensive diseases. The lethality rates differed greatly between specific disorders. The lethality rate was lower when scheduled maternity was located in a teaching hospital. CONCLUSION: Regarding these results it appears that the majority of obstetric patients with severe diseases are referred to suitable care, but a small proportion of women who had to change their type of care registered a significant higher lethality. PMID- 8706945 TI - Care of the patient with severe pregnancy induced hypertension. AB - Pregnancy induced hypertension remains the largest cause of maternal death in the UK. This is often related to inadequate or incorrect care. Recent randomised studies have enabled evidenced based clinical practice to be directed to the management of this condition. This allows a logical stepwise management structure to be developed in each hospital to make sure all women at risk get the best care available. This starts with comprehensive antenatal care with close collaboration between all professionals involved, early referral to an outpatient daycare unit if hypertension develops or is suspected, early use of antihypertensive therapy to control blood pressure once the diastolic is persistently above 100 mmHg, the use of magnesium sulphate to control convulsions, delivery on the best day in the best way and careful fluid management after delivery. Postnatal care should also contain risk assessment to allow directed counseling to the women concerning subsequent pregnancies. PMID- 8706946 TI - Serious haemorrhage during delivery. PMID- 8706948 TI - Training in the management of critical problems: teacher's view. AB - Teaching in critical obstetric problems should have special interest during residency and thereafter. Obstetric emergencies are relatively rare but may occur at any time. The obstetrician at that moment enters a special area requiring a multidisciplinary approach. From experiences in recent years and study of the literature the following recommendations can be summarized; (1) the need to understand (patho)physiologic changes in pregnancy, (2) cultivation of an anticipative attitude towards conditions with elevated risks, (3) adequate knowledge of diagnostic procedures, (4) the discipline to make a differential diagnosis, (5) experience with monitoring of (fetal and) maternal condition, (6) availability of management protocols for emergencies such as shock, eclampsia, uterine rupture, amniotic fluid embolism, thrombo-embolism, sepsis and diabetic ketoacidosis, (7) awareness of pitfalls with inspection of lesions and assessment of blood loss, (8) awareness that caesarean section without prior stabilization can be a life threatening procedure, (9) practice in life-saving measures such as uterine compression, packing, ligation of vessels, postpartum hysterectomy, (10) teaching of postoperative care, (11) insight into the cascade of events finally leading to multi-organ failure. Obstetric emergencies require a disciplined approach, in which teamwork is the cornerstone. PMID- 8706947 TI - Place of emergency arterial embolisation in obstetric haemorrhage about 16 personal cases. AB - Delivery bleedings which persist despite of an efficient medical treatment must be offered embolization without delay. Embolization is efficient in nearly 100% of cases; sometimes it has to be repeated. Embolization will it correct coagulopathy in few hours. It is a conservative treatment. It is nowadays undoubtedly the treatment of choice. Territorial organization for the hospital without interventional radiology is needed to organize the transfer of the bleeding patients without delay. Thus, mortality and morbidity as well as hysterectomy will not exist anymore. PMID- 8706949 TI - Training in the management of critical obstetric problems: trainee's view. AB - This paper evaluates the content of the training and the educational methods used in teaching how to handle critical obstetric problems. Structural training, description of aims and objectives and educational counselling encourage the acquisition, assessment and application of knowledge and skills. Educational counselling combined with continuous assessment can compensate for structural changes in the health service and improve and retain good clinical practise. PMID- 8706950 TI - The brain of the asphyxiated fetus--basic research. AB - The question whether asphyxia may be harmful to the brain of the fetus and newborn is largely related to the gestational age. Experimental studies on chronically prepared fetal sheep suggest that the resistance to asphyxia associated with maturity is not only related to structural changes in the fetal brain during development. Also, maturational changes of the sympathetic nervous system seem to be involved, which provide the neurohumoral basis for an effective circulatory and metabolic centralization to ensure intact survival of asphyxia (Jensen and Berger, 1991). This review will summarize the peculiarities of the fetal circulation, the redistribution of organ blood flow during hypoxaemia and asphyxia and its changes during development. Finally, it will focus on direct effects of asphyxia on the fetal brain. Future work in this field will have to concentrate on the important question how the fetal brain can be protected against the adverse effects of asphyxia. PMID- 8706951 TI - Paracrine mechanism of ovarian regulation. PMID- 8706952 TI - Predictive medicine. PMID- 8706953 TI - Maternal serum markers for fetal trisomy 21 screening. AB - A population at increased risk for fetal trisomy 21 can be defined by means of maternal serum markers. Various markers have been used since 1984, and the following have proved most valuable: hCG, free beta hCG, AFP, and estriol. Two prenatal screening periods should be distinguished: first trimester (8-14 weeks) and second trimester (14-18 weeks). Only the latter has been prospectively evaluated. In a prospective study, we assayed hCG in second trimester serum. A risk factor combining maternal age and hCG was defined and amniocentesis was offered to patients at increased risk for fetal trisomy 21. Out of 51,048 patients under 38 years of age, 135 had a trisomy 21-affected fetus. In 36,697 patients under 35, we observed 70 cases of trisomy 21, of which 41 (59%) were in the group at risk. Karyotyping was performed in 7.1% of these patients. In 11,351 patients aged 35-37 years, there were 65 cases of trisomy 21, of which 52 (80%) were in the group at risk. Karyotyping was performed in 26.8% of these patients. In our experience, parallel assaying of maternal serum AFP only detects a further 1% of trisomy 21 pregnancies for the same number of amniocenteses. These results confirm the findings of all previous prospective studies: maternal hCG screening is the most effective method of detecting trisomy 21 in the general population. PMID- 8706954 TI - Advantages and disadvantages of estrogen treatment by transdermal or oral administration. PMID- 8706955 TI - Mechanisms whereby oestrogens influence arterial health. AB - Coronary heart disease (CHD) is uncommon in premenopausal women compared with men of similar age, but its incidence increases after the menopause to reach that of men. There is now good population-based evidence that hormone replacement therapy (HRT) in postmenopausal women reduces the incidence of CHD, perhaps by up to 50%. Oestrogens have a beneficial effect on arterial health in many different ways. HRT may both reduce the risk of atheroma formation and improve arterial function. Depending on the formulation, HRT can lower LDL and triglycerides, and increase HDL. Oestrogen may also produce qualitative as well as quantitative improvements in lipoproteins. It can improve insulin resistance and hence carbohydrate metabolism, and may enhance fibrinolysis rather than coagulation. Thus these effects of HRT on risk factor for CHD will reduce the risk of atheroma development and progression. Oestrogen has direct effects on blood vessels and improves vascular function through various mechanisms including endothelium dependent and calcium-dependent processes. HRT should therefore now be considered for use in postmenopausal women with established CHD risk. PMID- 8706956 TI - Spontaneous evolution of intraepithelial lesions according to the grade and type of the implicated human papillomavirus (HPV). AB - Because of the fact that any meaningful classification should bear a close relationship to the biological behavior of the lesions, the usefulness of all new classifications of cervical precancer lesions can only be established by well controlled prospective follow-up studies. However, several methodological and conceptual problems are encountered in the natural history studies conducted during the past several decades. While reviewing the available prospective follow up studies on cervical intraepithelial neoplasia (CIN), Ostor (1993) found 3529 cases of CIN 1, of which 57% showed regression, persistence was found in 32%, progression to CIN III in 11%, and progression to invasive cancer in 1% of cases. The corresponding figures for CIN II were 43%, 35%, 22%, and 5%, respectively. The recognition of the association between human papillomavirus (HPV) and CIN has further complicated the assessment of the natural history of cervical precancer lesions. Results from the early prospective follow-up studies are remarkably consistent, however. Progression from HPV-NCIN (i.e., koilocytosis without CIN) to CIN I or greater was reported for 18 (8%) of 232 women followed by Syrjanen et al. for an average of 25 months, for 26 (8%) of 314 women followed by de Brux et al. (1981) for 15 to 18 months, and for 113 (13%) of 846 women followed for up to six years by Mitchell et al. During a 42-month follow-up period, 10% progression rate was found in 1269 women with HPV-CIN I, and in 17% of 762 women with HPV-CIN II by de Brux et al. (1983). The spontaneous regression rates were 53% and 39% in these cohorts, respectively. This is fully consonant with our experience from an almost 14-year follow-up of 530 women in Kuopio, where the spontaneous regression rate seems to increase in parallel with the extent of the follow-up time, currently being 66.7% for HPV-NCIN and 55.7% for HPV-CIN I. The figures for progression are 6.3% and 14.2%, respectively. It is obvious that the probability of a cervical precancer lesion to progress into an invasive disease increases with the severity of the atypia. Another distinct prognostic factor is HPV type, HPV 16 lesions possessing a significantly higher risk for progression than infections by other HPV types. The follow-up data also indicate, however, that even the high grade lesions may spontaneously regress, which should have important implications in therapy. The continuous problem still remains; these natural history observations only apply to a large series of women but are of little help in predicting the disease outcome in individual women. PMID- 8706957 TI - [In favor of annual screening for cancer of the cervix uteri]. PMID- 8706958 TI - Cytologic screening for cervix cancer: each year or each 3 years? AB - Screening is the systematic proposal of a medical procedure to a defined population at a defined interval. The efficacy and effectiveness of screening can only be demonstrated by a decreased mortality (not lethality) in case-control studies or randomised prospective studies. For cervix cancer, the only test with a decreased mortality proven by case-control studies (since 1979 only) is the cervix cytologic screening or 'Pap' smear. Systematic proposal is proven to be optimal every three years, no further gain being demonstrated for annual or biennial screening. PMID- 8706959 TI - Virologic screening. AB - Detection of high risk human papillomaviruses (HPV) has a high potential value for secondary prevention of cervical cancer. HPV testing can augment the sensitivity of conventional cytologic screening and can be used for triaging patients with a history of mild cytological abnormalities or low grade cervical intraepithelial neoplasia (CIN). Before screening for high risk HPVs can be generally recommended, refined commercially available HPV tests have to be established and their value has to shown in large epidemiologic studies. PMID- 8706960 TI - The colposcopic screening. AB - The objective of this paper is a valuation of colposcopy as a screening tool. As database, 392 patients with histologically confirmed intraepithelial neoplasia were used. Colposcopic and cytologic findings were compared with the final histologic diagnosis. The following results were obtained: (1)The colposcopic findings correlated with the histologic diagnosis to a significantly higher degree than the cytologic findings. Depending upon the rate of dysplasia, the colposcopic findings predicted the diagnosis in 84-97%. (2) The false-negative rate of cytology in condylomatous lesions and mild dysplasia was high (39 and 26%, respectively), in particular in comparison with the false-negative rate of colposcopy of 5% for both lesions. Thus, a negative smear does not exclude consistently a dysplasia of the cervix. (3) The false-negative rate of cytology for the high grade lesions (CIN II and CIN III) was 13 and 1% respectively and, thus, lower than in the low grade lesions. There were, however, considerable discrepancies in comparison with the histologic rating of the lesion. In CIN III cytology correlated with histology in only 61%, colposcopy, however, 85% (P < 0.001). Our results demonstrate that colposcopy is an excellent tool for detecting HPV caused lesions especially subclinical lesions and CIN I. Colposcopy is also a corrective for the false-negative cyto-smear rate (about 20-40%). Thus, colposcopy may be used as an effective quality assurance method and an excellent screening method in that colposcopy is superior in grading dysplastic lesions of the cervix. The application of the European terminology was advantageous. PMID- 8706961 TI - Preimplantation genetic diagnosis of cystic fibrosis (delta F508). AB - Cystic fibrosis is a common autosomal recessive condition caused by mutations in the cystic fibrosis transmembrane regulator gene. The major mutation is a three base pair deletion (delta F508). If both partners carry this deletion, the chance of having an affected child is 1 in 4. In vitro fertilization (IVF) with preimplantation genetic diagnosis allows the selection of the unaffected embryos only to be returned to the uterus. Preimplantation genetic diagnosis was attempted in 14 couples in which both partners carry the delta F508 deletion. A total of 22 cycles resulted in 170 normally fertilized embryos of which, 145 embryos were successfully biopsied and in 18 cycles, one or two unaffected embryos were transferred. A total of five clinical pregnancies established and at birth all five singletons have been confirmed as homozygous for the normal allele. From our experience, cleavage stage biopsy after in vitro fertilization provides sufficient embryos diagnosed as unaffected for transfer in this autosomal recessive disease. Also, pregnancy rates after the preimplantation diagnosis are similar to those with infertile couples. Prospects for applying preimplantation genetic diagnosis to autosomal dominant conditions, where incidences of having affected embryos would be higher, therefore, appear good. PMID- 8706962 TI - Indications for induction of labour. AB - Induction of labour is an important prophylactic procedure in the obstetric practice. Two major types of indications developed: induction because of advanced relevant obstetric complication, and the so called elective induction. PMID- 8706963 TI - Pelvic masses and endoscopic surgery: diagnosis. AB - Most adnexal masses are benign but a careful preoperative evaluation is necessary to exclude malignancy. Pelvic examination is considered to be of limited importance Ultrasonography is of prime importance and also tumor marker's assays. The last step of evaluation to exclude malignancy is the laparoscopic inspection of the tumor. If any signs of malignancy are found, laparotomy is performed immediately. The majority of patients with a benign adnexal cyst can be treated by operative endoscopy. PMID- 8706964 TI - Ovarian cysts. AB - The Author intends to compare the data available in literature on the topic of 'laparoscopic surgery of ovarian cysts' to his case material. From 1985 to 1994, the author carried out 920 laparoscopic operations for the removal of ovarian cysts. Of these, 13 were converted to laparotomies, mostly because of peri adnexal adhesions. There were 22 recurrences (endometriosic and mucinous multilocular cysts), and five severe complications (two purulent inflammations, one intra-operative haemorrhage and one post-operative one, one post-operative acute abdomen sine causa). In one case, an unrecognised endometrioid carcinoma was inadequately treated with laparoscopy. The author considers laparoscopy as the elective choice for surgical treatment of ovarian cysts. Suspect malignancy is not a contra-indication to laparoscopic surgery, since the removal of the operative specimen and its subsequent histological examination can be effected-in the large majority of cases-with the same results both laparoscopically and laparotomically. There is a problem with undiagnosed carcinomas in fertile females, but it is equally present in laparotomy. PMID- 8706965 TI - Uterine fibroids: place and modalities of laparoscopic treatment. AB - Only fibroids which give rise to symptoms resistant to properly conducted medical treatment and/or complicated myomas require surgical treatment. The possibility offered by the new surgical approaches enables myomectomies to be now carried out via laparoscopy. We report our technique of laparoscopic myomectomy performed since 1989. Monopolar coagulation is used for the uterine incision, after myomectomy, myometrium and serosa are sutured. Myomas are removed through the suprapubic puncture site after fragmentation of large myomas, through a colpotomy or using the morcellator. Our results enable us to say that, although it is a difficult technique, laparoscopic myomectomy is a safe and reliable technique even for large intramural myomas. PMID- 8706966 TI - The onset of labor. PMID- 8706967 TI - Helping the news media cover family planning. AB - To make informed choices about family planning, women and men need accurate information in the media as well as in the clinic. The media are looking for news important to the millions of people they reach each day. Thus helping the news media cover family planning fully and accurately merits the efforts of every family planning program. PMID- 8706968 TI - [Environmental pollution and infectious diseases. I. Isolation of a new serovar of the Salmonella genus, S.V 13.22:r: from a Periplaneta americana cockroach]. AB - The AA have isolated a new serovar of Genus Salmonella S.V 13.22:r:-, from a cockroach Periplaneta Americana, captured near the municipal slaughterhouse of Messina and near the Faculty of Veterinary Science. This fifth original report fift in the vast subject of the environmental pollution, where the cockroaches, as widely documented, unwind a primary importance part as spreader of Salmonella, too. PMID- 8706969 TI - Detection of chlamydia trachomatis antigen using the direct immunofluorescence technique: a four year survey in Northwestern Greece. AB - The prevalence of Chlamydia trachomatis antigen in patients visiting the University Hospital of Ioannina, North-western Greece during 1991-1993 is surveyed. Six hundred and twenty two (622) samples (scrapings from the cervix, uretra and conjunctiva) were examined for the detenction of C. trachomatis antigen using the direct immunofluorescence technique (DIF). From the 662 samples examined by DIF, 579 were found negative (93%) and 43 positive (7%). PMID- 8706970 TI - [Characteristics of a new Salmonella serovar: S. V 13, 22:r-, from S. bongori species]. AB - The A reports the characteristics of a new serovar of Salmonella: S.V 13, 22:r:-, isolated from a cockroach Periplaneta americana, all set against the background of ample technic notes. PMID- 8706971 TI - [Etiology and rational therapy of acute otitis media in adults]. AB - Acute otitis media (AOM) is an infection frequently found in children, but tends to be less frequent with age and its frequency in adults is only about 1%. The etiology of AOM in children is prevalently bacterial; numerous studies have shown the most common etiological agents. But the etiology in adults has not been well studied. The authors examined 40 cases of AOM in adults, the pathologic material was obtained by needle aspiration; in 32 samples there was bacterial growth. In the majority of the cases (94%) the bacteria isolated were the same as those found in children: S. pneumoniae, H. influenzae and B. catarrhalis; much rarer were S. pyogenes and S. aureus. On the potential beta-lactamase producing strains, this activity was measured. From our findings we believe that there is the necessity to have a rational antibiotic therapy (due to the difficulty in obtaining pathologic material) with active drugs for the probable etiological agents of AOM. PMID- 8706972 TI - [In vitro activity of several antibiotics on pathogenic micro-organisms isolated from the lower respiratory tract]. AB - Lower respiratory tract infections are clinically important, because of their high incidence, as well as the significant morbidity they cause. Hence the great importance to diagnose the etiology of these infections and treat the patient with selective therapy. In this study 367 bacteria isolated from lower respiratory tract were identified (253 Gram negative and 114 Gram positive) and tested towards augmentin, ceftriaxone, lomefloxacin, ciprofloxacin, ofloxacin and erythromycin. The in vitro activity of such antibiotics against the clinical isolates was compared. PMID- 8706973 TI - [Early diagnosis of HCMV infections in patients undergoing transplantation of the kidney]. AB - HCMV infection is a major cause of morbidity and mortality following kidney transplantation. Clinical diagnosis is difficult, and rapid and sensitive diagnostic methods are needed since antiviral therapy is available. One hundred forty-five consecutive kidney-transplanted patients were studied during a period of three months after transplantation. For laboratory diagnosis of HCMV infection, we looked for the presence of pp-65 antigen in polymorphonuclear leukocytes, HCMV-DNA and IgM. Demonstration of HCMV pp-65 antigen by immunofluorescence and HCMV DNA by PCR in leukocytes were efficient methods for early diagnosis of infection. PMID- 8706974 TI - Viral evasion of cytokine-mediated host defense mechanisms. PMID- 8706975 TI - [Protein p53 inhibits the activity of the enhancer of the immediate-early genes of murine cytomegalovirus]. AB - The protein encoded by the tumor suppressor gene p53 can complex and functionally interact with cytomegalovirus proteins produced during the immediate-early phase of infection. The functions of these complex are unclear but there is some evidence to suggest that binding of p53 to these viral proteins may inactivate p53 functions. Recent reports have shown that p53 is involved in regulation of transcription. In this study we have considered the possibility that p53 may regulate transcription of cytomegalovirus immediate early genes which play a crucial role for virus replication. Here we report that experiments in which NIH 3T3 cells were cotransfected with a p53 expression plasmid together with a reporter gene linked to the mouse cytomegalovirus immediate-early enhancer/promoter revealed that wild type p53 could efficiently reduce the transcriptional activity of this viral regulatory sequence. By contrast expression of a mutated p53 correlated with a much smaller reduction of transcription. Deletion mutants analysis of the enhancer revealed that repression of transcription by p53 requires a minimal promoter containing an SP1 consensus sequence and a TATA box. PMID- 8706976 TI - [Serum stimulates the transcriptional activity of the enhancer of the immediate early genes of the murine cytomegalovirus through p21 ras]. AB - The analysis of the MCMV IE enhancer revealed the presence of many putative binding sites for the transcription factors AP-1 and NFkB. Previous studies suggested that such factors represent a final target for the metabolic cascade triggered by serum and growth factors. On these basis we wanted to verify if serum stimulates the transcriptional activity of the MCMV IE enhancer through p21ras and AP-1 and NFkB according to the actual model of transduction of the mitogenic signal. Our data demonstrate that serum stimulates the MCMV IE enhancer through a pathway in which the p21ras is involded, as demonstrated by using the dominant inhibitory mutant ras(Asn 17). Moreover deletion mutant analysis of the enhancer showed that the serum responsive region lies between nucleotides -1280 and -285 and contains a high concentration of putative AP-1 and NFkB binding sites. PMID- 8706978 TI - [Epidemiologic study of infectious diarrhea in infants]. AB - The authors considered the epidemiological and microbiological aspects of the infectious diarrhea caused by bacteria, protozoan and viruses observed at the Infantile Hospital, Alessandria, and at the Malpighi Diagnostic Center, Chivasso, Turin, over a period of 14 months. Furthermore, a greater interest has been given to the role of Clostridium difficile diarrhea, since hospitalized patients are often predisposed to the development of this disease due to the antimicrobial prophylaxis or therapy. PMID- 8706977 TI - [The role of cytokines in viral infections]. AB - The immune response to viral infections is a combination of many events: viral antigen presentation to the immune system, proliferation of specific lymphocytes, cytokine intervention. This review describes the virus-cell relationship contributing to the latency state in viral infections (especially in AIDS) and cytokine network, its importance in the pathogenesis of viral diseases and its therapeutic potential. PMID- 8706979 TI - [Prevalence of urinary infections caused by GBS in diabetic patients]. AB - On a randomized sample of 305 diabetic patients and of 479 controls was studied for an epidemiological research about the colonization of urinary tract by Group B-Streptococci (GBS). The detection rate was 16.44% in the diabetics, and 8.77% in the controls. and it was higher in diabetic women than diabetic men (R.P. 1.96, p < 0.05). The age, the model of diabetes treatment, and the metabolic control of diabetic people didn't be significatively correlated with GBS presence. Most of GBS that have been identified in diabetic patients were belonged to the sierotype I, whereas in the controls were belonged to the sierotype III. PMID- 8706980 TI - [Stresses under conditions of a normal mode of life, during hypokinesia (simulating effects of weightlessness) and in space flights]. PMID- 8706981 TI - [Reflex constriction of resistance vessels in people with varying pathogenetic mechanisms of angina of effort]. PMID- 8706982 TI - [Dynamics of health in education students and teachers based on mathematical analysis of cardiac rhythm, anthropometric and psychophysiological parameters]. PMID- 8706983 TI - [Energy metabolism and autonomic function in students under study and examination stress]. PMID- 8706984 TI - [Effect of individual-adaptive psychophysiological correction on operator working capacity]. PMID- 8706985 TI - [Modern concepts on the mechanisms of aging]. PMID- 8706986 TI - [Sleep as an indicator of functional deviations in the body (based on materials of the XII Congress of the European Society of Sleep Research. May 22-27, 1994, Florence, Italy)]. PMID- 8706987 TI - [Effect of transcranial polarization on the state of supraspinal mechanisms of regulation of muscle tonus in patients with Parkinson's disease]. PMID- 8706988 TI - [Change in energy reserves during mental fatigue]. PMID- 8706989 TI - [Effect of the antistressor effect of acupuncture on the functional state of aqualung divers]. PMID- 8706990 TI - [A stochastic interpretation of Gustav Guillaume's psychotaxonomy (G. Guillaume "Principles of theoretical linguistics". Translated from the French. Edited by L. M. Skrelinaia. Moscow: Progress, 1992)]. PMID- 8706991 TI - [Cardiorespiratory system adaptation after surgical correction of intracardiac hemodynamics]. PMID- 8706992 TI - [Immunophysiologic mechanisms of adaptation to extreme conditions]. PMID- 8706993 TI - [Role of peptidergic mechanisms in thermoregulation]. PMID- 8706994 TI - [Stress-protective role of the opioidergic system under conditions of military operations]. PMID- 8706996 TI - [Adaptive functions of the emotions]. PMID- 8706995 TI - [Positron-emission tomographic study of human brain processing of various characteristics of visually presented words. I. Processing of visual properties of stimuli (color)]. PMID- 8706997 TI - [Analysis of the spatial organization of brain potentials during performance of a motor task]. PMID- 8706998 TI - [Role of cholinergic mechanisms in the functioning of specific structures of the visual analyzer]. PMID- 8707000 TI - [Effect of verbal reinforcement on visual-spatial function]. PMID- 8706999 TI - [Disorders in the perception of spatially oriented visual stimuli and Parkinsonism: role of the dopaminergic system]. PMID- 8707001 TI - [Sex differences in speech perception: effect of perceptive restoration of missing phonemes]. PMID- 8707002 TI - [Effect of postural muscle vibration on equilibrium maintenance in the frontal plane at various levels of stability]. PMID- 8707003 TI - [Lung expansibility and its assessment on the basis of the simplest models of the lung]. PMID- 8707004 TI - Mean levels and distributions of some cardiovascular risk factors in Italy in the 1970's and the 1980's. The Italian RIFLE Pooling Project. Risk factors and life expectancy. PMID- 8707005 TI - [Analysis of risk factors in a group of patients referred for heart transplantation for refractory heart failure: multivariate analysis]. AB - BACKGROUND: In the absence of evident clinical controindication heart transplantation can be performed in patients with critical degree of cardiac failure. The choice of adequate candidates for transplantation is frequently based on clinical severness but also influenced by factors as body size, blood group and so on. METHODS: To identify the presence of prognostic factors for patients with high risk of death, clinical, hemodynamic and therapeutic features of 70 patients (mean age 44.8 +/- 13.8) with critical cardiac failure (64 patients in IV class N.Y.H.A.) have been reviewed. Most of them were affected by dilated or ischemic cardiomyopathy. RESULTS: All patients were admitted to Cardiac Intensive Care Unit, 13 patients received transplantation, 24 were discharged and 33 died. Univariate analysis evidenced, that in the group of patients who died, sepsis (p = 0.0003), renal (p = 0.04) and liver function impairment (P = 0.007) occurred significantly more frequent. The differentiated use of catecholamines (P = 0.0007) and of vasodilators (P = 0.007), the need of mechanical ventilation (P = 0.0001) and the need of hemodialysis (P = 0.02) resulted significantly higher in the group of dead patients where mean arterial pressure (p = 0.01) and mean systemic resistances (P = 0.04) resulted significantly lower. With multivariate analysis were identified following independent risk factors of death: male sex, infections, mechanical ventilation, differentiated use of vasodilators, mean arterial pressure and mean pressure in pulmonary artery. CONCLUSIONS: As predictive risk factors of death in patients with critical cardiac failure and awaiting transplantation resulted: sex, the presence of infection, mechanical ventilation and differentiated use of vasodilators whereas hemodynamic parameters did not result of important predictive value. PMID- 8707006 TI - [Cardiac and vascular morphology in patients with "white-coat hypertension": echocardiographic and ultrasonographic study]. AB - BACKGROUND: The introduction of ambulatory blood pressure (ABPM) in the clinical practice has defined a new subgroup of hypertensive patients (pts) called "white coat" hypertensive pts. It has been reported that white coat hypertensive pts have less cardiac involvement than established hypertensive pts. This study was designed to examine the extent of cardiac and vascular involvement in pts with white coat hypertension and established hypertension. METHODS: Fifty-four previously never treated pts with mild or moderate hypertension underwent a 24h ABPM and an echocardiographic and vascular ultrasonographic study in order to assess left ventricular anatomical parameters and the intima-media thickness of common carotid arteries. Left ventricular dimensions and mass were obtained according to Penn convention. The intima-media thickness of the far wall of both common carotid arteries was measured at 5, 10 and 20 mm caudally to the flow divider; the average values were considered. RESULTS: Twenty-eight pts (age 43 +/ 8 years) had an average daytime ABPM below 140/90 mm Hg ("white coat") and 26 pts (age 46 +/- 10) had a consistently elevated diastolic blood pressure. Both groups had similar office blood pressure (152 +/- 9/101 +/- 7 vs 155 +/- 10/103 +/- 6 mm Hg) body surface mass (1.79 +/- 0.23 vs 1.86 +/- 0.21 mq), sex (18M/20F vs 18M/8F), duration of hypertension, glycolipidic parameters and smoking habit. The daytime ABPM was (by definition) significantly higher in established hypertensive pts than in white coat hypertensive pts (145 +/- 11/97 +/- 4 vs 129 +/- 7/83 +/- 5 mm Hg, p < 0.001). Left ventricular mass index and intima-media thickness resulted significantly greater in established (119 +/- 1.7 g/m2; 0.70 +/- 0.11 mm) than in white coat hypertensive pts (99 +/- 16 g/m2; 0.60 +/- 0.1 mm, p < 0.002). The prevalence of left ventricular hypertrophy and cardiac remodeling was more frequent in established hypertensive pts (53%) compared to white coat hypertensive pts (7.7%). CONCLUSION: The results of our study confirm that structural changes of left ventricle in white coat hypertensive pts are more limited than in established hypertensive pts and for the first time show that in white coat hypertensive pts the involvement of the conductance vessels is significantly lower than in established hypertensive pts. PMID- 8707007 TI - [Multiplane transesophageal echocardiography and adenosine in the study of coronary blood reserve]. AB - BACKGROUND: Coronary blood reserve is the capacity of coronary vessels to vasodilate and thereby to increase the blood flow, when the heart needs more energy. However, when a coronary stenosis occurs, the capacity to vasodilate is reduced or completely diminished. It is then necessary to use all the tools useful in evaluating the functional conditions of the coronary vessels. Above all, the intracoronary Doppler technique is used to measure the velocity of blood flow. Our purpose was to evaluate a non-invasive tool "Multiplane Transesophageal Echocardiography" in the study of velocity of the anterior descendent artery before and after adenosine infusion. METHODS: At first, we studied 28 patients (pts), which we divided in two groups: Group A, 18 pts 59.38 +/- 8.23 mean age, 15 M. and 3 F., with anterior descending disease; Group B 10 pts, 59.20 +/- 8.48 mean age, 7 M. and 3 F, without significant stenosis (> 75%). Echocardiography examinations were performed with a 5 MHz multiplane probe, connected to a 1000 Hewlett Parkard echocardiography. Before the test, Diazepam 1 mg i.v. and Lidocaine spray were administered to the patients. We introduced the transesophageal probe and after choosing the best position of the aortic short axis view, we studied the anterior descending artery and measured the maximum and mean diastolic and sistolic velocities (V.MAX D., V.MN.D., V.MAX S., V.MN.S.). RESULTS: Transesophageal echocardiography allowed us to study the anterior descending artery in 95% of pts. There were no side effects, except for one pt affected by severe bradicardia. In Group B there was an increase of the diastolic and sistolic velocity after adenosine infusion, resulting twice greater they the rest values. The adenosine/rest velocities ratios were statistically significant (V.MAX D. p < 0.02) (V.MN.D. p < 0.03). CONCLUSION: Our results demonstrated an higher capacity of the Multiplane Transesophageal Echocardiography in studiing coronary blood reserve. We used adenosine, as a vasodilator drug, because of its short half-life and because it can be replatedly infused. The flow velocity values increased up to more than twice the rest values only in normal subjects. PMID- 8707008 TI - [Combined valve repair for mitral-aortic insufficiency]. AB - Reconstructive surgery is considered the best treatment of mitral insufficiency. Aortic valvuloplasty has been used only rarely in the past for the treatment of aortic insufficiency. Between October 1994 and November 1994, two patients admitted at our institution with the diagnosis of mitral and aortic insufficiency have been successfully submitted to combined valvuloplasty. Although long term results of aortic reconstruction are unavailable, short-term and middle-term results are satisfactory and warrant conservative surgery in selected cases. Aortic repair is an attractive option especially in those patients with combined aortic insufficiency and mitral disease suitable for valvuloplasty. PMID- 8707009 TI - [Surgical or percutaneous revascularization in multi-vessel coronary disease]. PMID- 8707010 TI - [Myocardial infarction and nitric oxide]. PMID- 8707011 TI - [Circulatory assistance in cardiac decompensation: when and how]. PMID- 8707012 TI - [Pharmacologic tests in cardiac decompensation]. PMID- 8707013 TI - [Genetics and molecular biology of cardiac hypertrophy]. PMID- 8707014 TI - [Intravascular echography: a new approach to the pathophysiologic assessment of atherosclerotic lesions]. PMID- 8707015 TI - [Management of the patient with ventricular extrasystole]. PMID- 8707016 TI - [Morphofunctional features of left ventricular hypertrophy]. PMID- 8707017 TI - [Therapy of myocarditis]. PMID- 8707018 TI - [Nuclear cardiology: techniques of metabolic evaluation]. PMID- 8707019 TI - [Inflammation and atherosclerosis]. PMID- 8707020 TI - [The use of transesophageal echocardiography in cardioversion for atrial fibrillation]. PMID- 8707021 TI - [Physician-patient relations in cardiology]. PMID- 8707022 TI - [Propafenone and sotalol: long-term efficacy and tolerability in the prevention of paroxysmal atrial fibrillation. A placebo-controlled double-blind study]. AB - BACKGROUND: Atrial fibrillation is a relatively frequent atrial arrhythmias activated with increased morbidity and mortality. METHODS: To assess the propafenone and sotalol efficacy in the prevention of paroxysmal atrial fibrillation (FA) we enrolled, in a double blind placebo controlled study over 1 year, 300 patients (168 males); mean age 52.3 +/- 17.2 years, randomized to receive orally, three times daily, either propafenone (mean daily dose of 13 +/- 1.5 mg/Kg; Group A: 102 patients) or sotalol (mean daily dose of 3 +/- 0.4 mg/Kg; Group B: 106 patients) or placebo (Group C: 92 patients). All subjects experienced in previous 12 months at least 4 FA episodes. During follow-up we considered atrial tachyarrhythmia (TAA) onset: FA recurrences and/or the onset of atrial flutter (FIA). Three patients (3%) of Group A and 5 (5%) of Group B interrupted therapy for side effects; 5 patients (5.5%) of Group C with supraventricular tachycardia interrupted the double blind therapy; 11 were lost to follow-up. RESULTS: Of the remaining 276 patients, TAA were observed in 43 (44.8%) of 96 patients in Group A, 28 (29.5%) of 95 patients in Group B and 62 (72.9%) of 85 patients in Group C. TAA were significantly less in A and B groups than in Group C (p < 0.005); a significant TAA reduction was also observed in patients treated with sotalol compared with those treated with propafenone (p < 0.05). TAA were: FA-118 (88.7%) and FIA-15 (11.3%). The arrhythmia free time was significantly shorter in Group C. CONCLUSIONS: Sotalol seems to be more effective than propafenone and therefore represents a valid alternative for FA prevention. PMID- 8707023 TI - [Radiofrequency perforation in pulmonary valve atresia and intact ventricular septum]. AB - METHODS: Four neonates with Pulmonary Atresia and intact Ventricular Septum underwent Radiofrequency Valvotomy in our institution from October 1994 to June 1995. All patients had "membranous" atresia with confluent and normal size pulmonary branches; one of them had abnormal tricuspid tensor apparatus and severe right ventricular outflow tract hypertrophy. The right ventricle was considered of sufficient size to support biventricular circulation in all cases. In three patients the pulmonary arteries were supplied by the arterial duct maintained open with intravenous infusion of Prostaglandins. The other patient had previously undergone a modified left Blalock Taussig shunt in another institution. The cardiac catheterization was performed by the femoral venous and arterial routes, under general anesthesia. After delineating the atretic valve with angiography, a 5 French right Judkins catheter was placed in the right ventricular outflow tract just beneath the membrane. A 2 french radiofrequency catheter (Cerablate PA 120 Osypka) was then passed through the right Judkins; with a mean energy of 5-10 watts over 3-5 seconds the valve was perforated. After a predilation with a 2,5 mm balloon catheter (New Probe USCI or Cobra SCIMED), the valve was dilated with balloon catheters 20 to 30% larger than the anulus (Cristal Balloon BALT). RESULTS: In all cases the procedure was successful without any complication; a significant decrease of right ventricular pressure and an excellent antegrade flow across the valve were achieved. Two patients were weaned from prostaglandins by sixth and eighth day after the procedure. The patient with previous Bialock Taussig shunt showed a complete recovery of the right ventricle, but four months later underwent surgery for shunt closure and enlargement of the left pulmonary branch. These three patients are well at latest follow-up (5-10 months), with normal biventricular circulation. The other patient remained duct-dependent and was operated on of outflow patch and shunt nine days after the procedure, with good result; he unfortunately died the tenth postoperative day for complications of hemopericardium related to epicardial wires removal. CONCLUSIONS: In our opinion radiofrequency valvotomy is the first choice procedure in cases suitable for biventricular repair. In selected patients it may represent a definitive treatment. PMID- 8707024 TI - [Factors limiting adaptation to exercise after the Fontan operation: role of altered pulmonary function]. AB - BACKGROUNDS: The abnormal adaptation to exercise after Fontan operation, is primarily due to a suboptimal increase in cardiac output. We postulated that abnormal pulmonary function could cooperate in reducing effort tolerance of these patients (pts). METHODS: We have evaluated 10 pts, mean age 15 +/- 8 yrs, with a mean postoperative follow-up of 6 +/- 2 yrs. Seven pts were asymptomatic while 3 had mildly decreased effort tolerance. These pts were evaluated with echocardiogram, cycle ergometer stress test, basal and effort spirometry and perfusion lungs scanning. RESULTS: Basal ejection fraction was normal in 9 pts and mildly depressed in 1 pt. Basal cardiac index was 1.8 + 2.2 l/min/m2 and increased to 4.8 + 5 l/min/m2 during exercise. Effort tolerance was 73% +/- 21% of predicted values, with maximal O2 consumption of 60% +/- 13% of predicted. Basal spirometry showed a mild restrictive pattern in 7 pts with a decrease in total pulmonary capacity and in forced vital capacity (mean values respectively 79% +/- 14% and 75% +/- 18% of predicted), otherwise respiratory reserve resulted normal in all pts. Carbon monoxide diffusion was impaired in all pts with a mean value of 49% +/- 10% of predicted. O2 saturation near normal in basal condition decrease during exercise (mean value 96% +/- 2% and 91% +/- 7%). The VE/VO2 and Vd/Vt ratio were both abnormally increased (respectively 111% +/- 28% and 186% +/ 39% of predicted). Lung scanning was abnormal in 6 pts with defects of perfusion localized prevalently in left lung (5/6) especially in upper lobe (4/6). CONCLUSIONS: After Fontan operation exercise tolerance is reduced as consequence of 1) a suboptimal increase in cardiac output and 2) abnormal gas diffusion probably due to a damage of alveolo-capillary membrane and/or to a ventilation/perfusion mismatch as showed by abnormalities in lungs perfusion and by increase Vd/Vt ratio. PMID- 8707025 TI - [Efficacy and tolerability of felodipine and nifedipine in stable angina refractory to beta-blocker therapy]. AB - BACKGROUND: Medical therapy of stable angina contemplates beta-blockers as a first line. The combination of dihydropyridines with beta-blocking drugs enhances the effectiveness of both single therapies. Nifedipine, in the usual formulation (AR), is burdened by an unsatisfactory tolerability, and this is the main reason to study new dihydropyridines. AIM: To compare efficacy and tolerability of felodipine ER 10 mg o.d. with that of nifedipine AR 20 mg b.d. in patients with stable angina pectoris refractory to beta-blocker therapy. PATIENTS AND METHODS: Of 15 initial patients, 14 were entirely evaluable for the study, the design of which was double blind, double dummy, random cross over and placebo controlled. All patients showed reproducible threshold of ischemia at exercise testing. In constancy of beta-blocker therapy, they were given placebos for 2 weeks, then one of the active drugs with a placebo of the other one for 4 weeks, followed by the cross-over period of 4 weeks. Efficacy and tolerability of treatments were evaluated by clinical observation and rest and exercise radionuclide angiography. At the end of each individual study, it was decided blindly if and which of the 2 drugs seemed preferable, considering symptoms, undesired collateral effects and the results of exercise procedures. RESULTS: The efficacy on angina of the 2 active treatments was not different. More patients suffered undesired side effects on nifedipine than on felodipine. Left ventricular ejection fraction (LVEF) at rest was 65.3 +/- 4.3% (s.e.) on placebo, 64.6 +/- 2.6% on felodipine and 67.5 +/- 2.5% on nifedipine (p n.s.). A significant reduction in resting LV function (that is, a decrease of LVEF > or = 5%) was observed in 2 patients on felodipine and 3 on nifedipine, but in both groups other 3 patients showed improvement in LVEF. During exercise, LVEF decreased 6.1 +/- 2.0% on placebo and 3.3 +/- 3.2% on nifedipine, while it increased 1.0 +/- 2.6% on felodipine (p < 0.01 vs. placebo). At the end of the study, felodipine was blindly judged superior to nifedipine in 10 patients, nifedipine was superior in 1 case, in the other 3 there was no clear difference (p < 0.02). CONCLUSIONS: In 14 patients with stable angina refractory to beta-blockers, the addition of felodipine or nifedipine has similar antiischemic effects. However, felodipine showed better results in LVEF response to exercise and less side effects, and this leaded to a more frequent blind choice of felodipine versus nifedipine to add to beta-blocker therapy. PMID- 8707026 TI - [Role of a "bystander" concealed accessory pathway in the response to verapamil and flecainide treatment in a patient with nodal reentry tachycardia]. AB - The possibility of multiple accessory atrioventricular pathways as that of multiple atrioventricular "nodal" pathways represents a relatively usual finding in patients suffering from supraventricular reciprocating tachycardias; the occurrence of both double atrioventricular nodal and concealed accessory pathways in the same patient, on the contrary, is far less common, and is usually an unsuspected condition prior to an electrophysiologic endocavitary study. The authors describe a patient suffering from paroxysmal tachycardias of both types, which have been clearly demonstrated by a transesophageal electrophysiological study. The report shows the different effects of two drugs, verapamil and flecainide, on these reciprocating circuits. Flecainide showed an immediate effectiveness in this condition, while verapamil, although very effective for the "nodal" reciprocating tachycardia, might worsen the accessory reentrant one, and cause a "jump" from the first to the second form. PMID- 8707027 TI - Acute myocardial infarction and residual impairment of coronary perfusion: unusual complications of mitral endocarditis in a paediatric patient. AB - In children, acute myocardial infarction is a rare complication of infective endocarditis. Since it is usually fatal, few data on the patients who have survived are available. We report the case of a child with acute myocardial infarction and residual impairment of the coronary reserve, complicating a mitral endocarditis. PMID- 8707028 TI - [Heart surgery, cardiopulmonary bypass and inflammatory response. I. Changes in hemostasis and complement]. AB - The use of cardiopulmonary bypass for surgical cardiac procedures is characterized by a whole-body inflammatory reaction due to the contact of blood through nonendothelialized surfaces; this stimulates the organism to recognize the cardiopulmonary bypass system as "nonself" and to activate specific (immune) and nonspecific (inflammatory) responses. These responses are then related with postoperative damage to many body systems, like pulmonary, renal or brain dysfunction, excessive bleeding and postoperative sepsis. In this paper, present knowledge on untoward responses of the patient to the use of cardiopulmonary bypass in cardiac surgery is reviewed and discussed, particularly focusing on the perturbation of the hemostasis and of the complement activation system. PMID- 8707030 TI - [Not only DRG (without new "Quality" the recently introduced hospital financing system risk failure)]. PMID- 8707029 TI - [Echocardiography with pharmacological stress: dobutamine or dipyridamole?]. PMID- 8707031 TI - [Can the numerical and statistical strength of a mega trial with an intervention protocol wrong from the physiopathologic viewpoint lead to definitive conclusions?]. PMID- 8707032 TI - [Mitral valve insufficiency in asymptomatic patients: when to think about reparative surgical intervention, when to decide about replacement]. PMID- 8707033 TI - [Thyroid gland dysfunctions and disorders in pregnancy and puerperium. Prevention, diagnosis and therapy]. PMID- 8707034 TI - [History of the European hospital to 1800]. PMID- 8707035 TI - [Psychological coping with abortion by the woman]. PMID- 8707036 TI - [Legal concerns. Healing treatment, healing trial and human experiment]. PMID- 8707037 TI - [Diagnostic and clinical value of Doppler ultrasound in obstetrics]. AB - The diagnostic value (efficiency) of Doppler ultrasound in obstetrics is assessed by studying the association or correlation between Doppler measurement parameters and pregnancy outcome parameters, without influencing the clinical management. The clinical value (efficacy) is however assessed by studying the ability to improve the outcome of pregnancy if Doppler ultrasound is used in obstetrical management for the purpose of possible intervention. The accepted method for assessment of the diagnostic value is the observational study, and the best method for the assessment of the clinical value is the randomised controlled management study (clinical trial). Observational studies concerning the diagnostic value have been widely reported in the literature. According to these studies significant relationships between abnormal Doppler findings and the following obstetrical problems were found: intrauterine growth retardation, preterm delivery, acidosis determined by fetal blood sampling, abnormal intrapartum fetal heart rate, neonatal depression and acidosis, and admission to neonatal intensive care unit. However from these studies it cannot be concluded that pregnancy outcome will be improved by using this information clinically. This problem can only be properly addressed by appropriate clinical management studies. The majority of the published randomised controlled trials showed beneficial effect from Doppler ultrasound application in high-risk pregnancy management. These effects included a significant decrease in the duration of antenatal and neonatal hospital stay, the rate of labour induction, and the rate of emergency Caesarean section. At the same time there was no difference in the gestational age at delivery or in the total Caesarean section rate. None of these trials alone was large enough to demonstrate a statistically significant reduction in perinatal mortality. However cumulative meta-analysis allows pooling to evaluate the results from separate but similar studies. The results from such an analysis showed that in high-risk pregnancies managed with Doppler there is a significant decrease of 50 per cent in perinatal mortality and in stillbirths of anatomically normal fetuses. It is important to note that there was no increase in neonatal or maternal morbidity associated with the use of Doppler ultrasound, and that the reduction in perinatal mortality was not the result of a delay in timing fetal death. PMID- 8707038 TI - [Normal values for relative Doppler indices. A/B ratio, resistance index and pulsatility index of the uterine artery and umbilical artery in normal pregnancy. A longitudinal study]. AB - 37 women (of a total of 50) with an uneventful obstetric history, normal course of a singleton pregnancy and spontaneous delivery at term (> 37 weeks, birthweight > 10th centile) were studied at two to four weeks intervals in a longitudinal design using cw-Doppler velocimetry (Doptek 9013,4MHz, SPTA 30 mWcm2, high pass filter 200 Hz, Chichester, U.K.) to establish normal reference values for Doppler flow velocity waveform indices (DI) A/B ratio, RI, PI obtained from uterine and umbilical arteries from 16 to 40 weeks of gestation. Readings from the placental (p) and non-placental (np) uterine and umbilical arteries formed the database from which percentiles (5th, 50th, 95th) were calculated for weeks 16, 20, 24, 28, 32, 36 and 40 of gestation. DI consistently decreased from week 16 to term. For the uterine arteries this decrease was significant from weeks 16 to 24 with a further but insignificant decrease from 24 weeks to term. Up to 24 weeks DI values obtained from the placental side were significantly lower than those from the non-placental side. DI values from the umbilical arteries decreased continuously from 16 weeks to term with a significantly negative correlation DI values to gestational age. PMID- 8707039 TI - [Subpartal diagnosis of umbilical cord encirclement using color-coded Doppler ultrasonography and correlation with cardiotocographic changes during labor]. AB - Umbilical cord complications are the most common cause of pathologic fetal heart tones during delivery. The inauguration of colour-coded Doppler ultrasound in obstetrics has made the definite diagnosis of umbilical cord encirclement during delivery possible. The prospective study introduced here examines the question of how exactly an encirclement can be seen by Doppler during delivery, its influence on cardiotocographic results, delivery mode, and fetal outcome. 107 patients in labour with cervical dilatation were examined in a prospective study using colour coded Doppler ultrasound to determine cases of umbilical cord encirclement. In 50 cases, encirclement could be visualised, 48 of which were confirmed post partum. Encirclement could be ruled out in 57 other cases. A sensitivity of 96% and specificity of 100% resulted. No significant differences could be found with regard to mode of delivery and fetal outcome. However, the umbilical cord in cases of encirclement was significantly longer than when no encirclement occurred. Assessment of fetal heart tones demonstrated a significantly higher rate of variable decelerations in the patient group with umbilical cord encirclement compared to that without. In conclusion, our results show that the early diagnosis of umbilical cord encirclement during delivery allows appropriate assessment of fetal heart tone changes,justifying temporising management under continuous monitoring with possible micro-blood analysis. PMID- 8707040 TI - [Intensive monitoring of the fetus sub partu--value and false interpretation]. AB - The past 30 years have seen dramatic advances concerning safety of the unborn child unsurpassed during 2400 years before, i.e. since the time of Hippocrates. Part of these advances is fetal monitoring during labor. It has recently been recognized that severe brain damage due to intrapartum fetal hypoxia is more rare than previously assumed. Legal consequences are to be expected. Expert opinion ought to investigate and judge the cause of possible intrapartum fetal damage more thoroughly and critically than it was customary. It is unfortunate that some authors in the framework of this development and for other reasons as well have come to the erroneous conclusion that the intensive intrapartum fetal monitoring is of less significance than previously believed. Such opinions are slanted and scientifically untenable. Intensive intrapartum fetal monitoring fulfills also the important task of reducing the early acidosis related to morbidity which has to be taken seriously since it may affect nearly all important organ systems of the fetus and the newborn. Intrapartum fetal monitoring has to (correction for o) be considered of great value regard to modern medical knowledge. PMID- 8707041 TI - [Clinical results of pulse oximetry sub partu for measuring fetal arterial oxygen saturation]. AB - By means of reflexion pulse oximetry the arterial oxygen saturation (SpO2) of 100 fetuses intrapartum was measured in a high-risk group and compared with clinical parameters. There is a connection between the duration of SpO2 measurements below 30%, the delivery mode and reduced pH-values in the umbilical artery measured post partum. SpO2 values below 30% also occur temporarily with uneventful deliveries but they occur with increased frequency within the group of operatively delivered children with a duration of more than 20 minutes. On the other hand the group with SpO2 values ranging between 40 and 60% showed less operative deliveries and a better condition of the newborn (Apgar, pH-metry). If prognostically unfavourable additional criteria with variable decelerations increase, a tendency towards decreasing fetal oxygen saturation values is recognisable. In the case of suspicious heart rate patterns or patterns difficult to assess the continuous registration of saturation values is both medically and forensically of inestimable value. There were no increases in fetal morbidity or signs of amnion infection syndrome. By means of a new sensor, a longer registration time with a reliable signal quality is achieved, so that assessment of the oxygen saturation in relation to labour activity becomes possible. PMID- 8707042 TI - [Magnetocardiography for assessment of fetal heart actions]. AB - Magnetocardiography provides a new, alternative method for fetal surveillance. This method allows the registration of changes in the magnetic field produced by the fetal cardiac system. In comparison to fetal ECG, the fetal magnetocardiogram (fMCG) is not affected by intermediate tissue, in particular by the insulating effect of the vernix caseosa. Furthermore fMCG's with a good signal noise ratio can be obtained in the second trimester. A multichannel biomagnetometer allows the non-invasive measurement of the magnetic signals produced by the fetus. Examples of dta and evaluation techniques are given. Signal processing software allows the identification of all fetal beats in a data set, permitting the calculation of an averaged beat and the evaluation of signal morphology and cardiac time intervals. Furthermore it is possible to produce a beat to beat time series with high temporal resolution, making the evaluation of heart rate variability with measures from the time, frequency, and phase domain accessible. Particularly the latter are of interest in investigating changes in temporal complexity of the fetal cardiac system. Magnetocardiography thus holds the promise of improved antenatal surveillance as of the second trimester. PMID- 8707043 TI - [Gerontologic aspects of postmenopausal measurement of bone density and prevention of osteoporosis]. AB - I. Among 454 postmenopausal women to undergo bone mineral density (BMD) examination 7 out of 10 were aged 50 to 70 years. 1. 50 of patients older than 70 years had BMD examination because of being clinically suspicious of osteoporosis. 2. In this group of age 4 out of 10 women had BMD results of the lumbar spine below 0,8 gcm2. 3. Half of all women aged up to 50 years had a positive family history of osteoporosis. 4. Patients older than 60 years often underestimated their own risk of osteoporosis. 5. 50 of women up to the age of 50 and only one sixth of women older than 70 said they had very good to good knowledge of osteoporosis. 6. Resentments against estrogen substitution are rapidly growing with every decade of age. 7. Only one third of the women selected by BMD indication were ready to accept longterm estrogen substitution to prevent osteoporosis. II. 4 out of 10 women had entered menopause maximally 8 years ago.1. Out of this group 6 of 10 patients were substituted with estrogen. 2. When menstruation had sisted more than 15 years ago the BMD examination indication, clinically suspicious of osteoporosis, rose immensely and made up for 50 of patients. 3. One third to one half of patients who had reached menopause more than 22 years ago showed pathological BMD results, which means they were at risk of fractures. 4. In this group only 1 in 10 women had a positive history of osteoporosis. III. Obese postmenopausal women (more than 25 of overweight according to Broca) had pathologic BMD results only half as often as postmenopausal women with normal weight. Summing it up these facts may be the basis of special counselling on the prevention of osteoporosis by estrogen substitution. PMID- 8707044 TI - [Climacteric arthralgia and hand arthrosis]. AB - This study aimed at a possible classification of climacteric arthroses of the hand. 132 females suffering from arthralgias of the wrist and finger joints who had consulted our outpatient department originally because of climacteric complaints, were examined while asking them to complete a questionnaire with clinical criteria for classifying arthroses of the hand. The requisite number of criteria required for classification as ?arthrosis of the hand? were attained in all three groups of females (premenopausal and postmenopausal women without operation, as well as hysterectomised and/or ovarectomised women?, although the ex-ray showed no signs of degenerative arthritis. The subjective complaints voiced by the patients are opposed to the x-ray finding, so that it is not permissible to classify climacteric arthralgias of the wrist and finger joints as ?arthritis?. PMID- 8707045 TI - [Hip diseases in pregnancy]. AB - Transient osteoporosis and avascular femoral head necrosis represent rare but typical complications in the third trimenon of pregnancy. On the basis of five cases we report on the diagnosis, course and treatment of these diseases which are rarely mentioned in German literature. PMID- 8707046 TI - [Doppler sonography of the ductus venosus in high risk pregnancies at 23 and 26 weeks]. AB - Ductus venosus blood flow was assessed in 3 high-risk pregnancies, which were terminated at 23 to 26 weeks' gestation because of HELLP-syndrome. The fetuses were extremely growth-retarded and showed diastolic reverse flow in the umbilical artery. In all cases the ductus venosus flow pattern was abnormal and finally showed loss of blood flow during atrial contraction. This finding suggests that the fetal central venous pressure is pathologically increased and may be interpreted as a sign of threatening cardiovascular decompensation. PMID- 8707047 TI - Fluorescent Proteins and Applications. Symposium proceedings. Palo Alto, California, USA, 6-7 March 1995. PMID- 8707048 TI - Application of novel vectors for GFP-tagging of proteins to study microtubule associated proteins. AB - We describe the construction of pBact-NGFP and pBact-CGFP, two expression vectors that incorporate green fluorescent protein (GFP) as a fluorescent tag at the N- or C terminus of the produced protein. When transfected into recipient cells, GFP tagged proteins can be visualised in the living cells using standard fluorescence microscopy techniques. Using these expression vectors, we have produced GFP tagged versions of the neuronal microtubule-associated proteins (MAP), MAP2c and Tau34, in a number of different cell types. Both GFP-MAP2c and GFP-Tau34 were fluorescent and retained their ability to bind to microtubules. The pBact-NGFP and pBact-CGFP expression vectors represent a fast and convenient way to produce fluorescently tagged polypeptides of selected sequences encoding whole proteins or fragments for the analysis of function and dynamic events in living cells. PMID- 8707049 TI - Targeting aequorin and green fluorescent protein to intracellular organelles. AB - Two proteins of Aequorea victoria were molecularly engineered and produced in mammalian cells, in order to serve as specific reporters of subcellular microenvironments. Aequorin (AEQ), a Ca(2+)-sensitive photoprotein, was successfully targeted to three intracellular locations: cytosol, nucleus and mitochondria. The recombinant apoprotein, reconstituted into active AEQ by the addition of the prosthetic group to the culture medium, allows the direct measurement of [Ca2+] within those compartments, thus directly addressing questions of large biological interest. The same approach was utilized for the green fluorescent protein (GFP) for specific labelling, in vivo, of the various subcellular structures. GFP was targeted to mitochondria: the recombinant protein, strongly fluorescent in a highly reducing environment, provides a powerful tool for visualizing these organelles in living cells, and may represent the prototype of a new family of intracellularly targeted fluorescent probes. PMID- 8707050 TI - Fluorescence resonance energy transfer between blue-emitting and red-shifted excitation derivatives of the green fluorescent protein. AB - We report fluorescent resonance energy transfer (FRET) between two linked variants of the green fluorescent protein (GFP). The C terminus of a red-shifted variant of GFP (RSGFP4) is fused to a flexible polypeptide linker containing a Factor X a protease cleavage site. The C terminus of this linker is in turn fused to the N terminus of a blue variant of GFP (BFP5). The gene product has spectral properties that suggest energy transfer is occurring from BFP5 to RSGFP4. Upon incubation with Factor X(a), the protein is cleaved, and the two fluorescent proteins dissociate. This is accompanied by a marked decrease in energy transfer. The RSGFP4::BFP5 fusion protein demonstrates the feasibility of using FRET between two GFP derivatives as a tool to monitor protein-protein interactions; in addition, this construct may find applications as an intracellular screen for protease inhibitors. PMID- 8707051 TI - Dual color microscopic imagery of cells expressing the green fluorescent protein and a red-shifted variant. AB - The green fluorescent protein (GFP) from the jellyfish, Aequorea victoria, has become a versatile reporter for monitoring gene expression and protein localization in a variety of cells and organisms. GFP emits bright green light (lambda max = 510 nm) when excited with ultraviolet (UV) or blue light (lambda max = 395 nm, minor peak at 470 nm). The chromophore in GFP is intrinsic to the primary structure of the protein, and fluorescence from GFP does not require additional gene products, substrates or other factors. GFP fluorescence is stable, species-independent and can be monitored noninvasively using the techniques of fluorescence microscopy and flow cytometry [Chalfie et al., Science 263 (1994) 802-805; Stearns, Curr. Biol. 5 (1995) 262-264]. The protein appears to undergo an autocatalytic reaction to create the fluorophore [Heim et al., Proc. Natl. Acad. Sci. USA 91 (1994) 12501-12504] in a process involving cyclization of a Tyr66 aa residue. Recently [Delagrave et al., Bio/Technology 13 (1995) 151-154], a combinatorial mutagenic strategy was targeted at aa 64 through 69, which spans the chromophore of A. victoria GFP, yielding a number of different mutants with red-shifted fluorescence excitation spectra. One of these, RSGFP4, retains the characteristic green emission spectra (lambda max = 505 nm), but has a single excitation peak (lambda max = 490 nm). The fluorescence properties of RSGFP4 are similar to those of another naturally occurring GFP from the sea pansy, Renilla reniformis [Ward and Cormier, Photobiochem. Photobiol. 27 (1978) 389-396]. In the present study, we demonstrate by fluorescence microscopy that selective excitation of A. victoria GFP and RSGFP4 allows for spectral separation of each fluorescent signal, and provides the means to image these signals independently in a mixed population of bacteria or mammalian cells. PMID- 8707052 TI - Intravital imaging of green fluorescent protein using two-photon laser-scanning microscopy. AB - Imaging a fluorophore in a living tissue presents several unique problems. The fluorescence from the labeled cell(s) may be weak, the labeled cells may be buried deep within tissue and the presence of a fluorophore may render the cells photo-sensitive. Two-photon laser-scanning microscopy (TPLSM) offers several advantages in meeting these challenges. We show that TPLSM provides greater sensitivity, better resolution and less photo-bleaching, as compared to confocal laser-scanning microscopy. The dramatically reduced photo-bleaching makes it possible to image cells continuously for long periods of time. Therefore, TPLSM allows a safer and higher-resolution means of imaging living cells labeled with a variety of fluorophores, including green fluorescent protein. PMID- 8707053 TI - FACS-optimized mutants of the green fluorescent protein (GFP). AB - We have constructed a library in Escherichia coli of mutant gfp genes (encoding green fluorescent protein, GFP) expressed from a tightly regulated inducible promoter. We introduced random amino acid (aa) substitutions in the twenty aa flanking the chromophore Ser-Tyr-Gly sequence at aa 65-67. We then used fluorescence-activated cell sorting (FACS) to select variants of GFP that fluoresce between 20-and 35-fold more intensely than wild type (wt), when excited at 488 nm. Sequence analysis reveals three classes of aa substitutions in GFP. All three classes of mutant proteins have highly shifted excitation maxima. In addition, when produced in E. coli, the folding of the mutant proteins is more efficient than folding of wt GFP. These two properties contribute to a greatly increased (100-fold) fluorescence intensity, making the mutants useful for a number of applications. PMID- 8707054 TI - Deletion mapping of the Aequorea victoria green fluorescent protein. AB - Aequorea victoria green fluorescent protein (GFP) is a promising fluorescent marker which is active in a diverse array of prokaryotic and eukaryotic organisms. A key feature underlying the versatility of GFP is its capacity to undergo heterocyclic chromophore formation by cyclization of a tripeptide present in its primary sequence and thereby acquiring fluorescent activity in a variety of intracellular environments. In order to define further the primary structure requirements for chromophore formation and fluorescence in GFP, a series of N- and C-terminal GFP deletion variant expression vectors were created using the polymerase chain reaction. Scanning spectrofluorometric analyses of crude soluble protein extracts derived from eleven GFP expression constructs revealed that amino acid (aa) residues 2-232, of a total of 238 aa in the native protein, were required for the characteristic emission and absorption spectra of native GFP. Heterocyclic chromophore formation was assayed by comparing the absorption spectrum of GFP deletion variants over the 300-500-nm range to the absorption spectra of full-length GFP and GFP deletion variants missing the chromophore substrate domain from the primary sequence. GFP deletion variants lacking fluorescent activity showed no evidence of heterocyclic ring structure formation when the soluble extracts of their bacterial expression hosts were studied at pH 7.9. These observations suggest that the primary structure requirements for the fluorescent activity of GFP are relatively extensive and are compatible with the view that much of the primary structure serves an autocatalytic function. PMID- 8707055 TI - Applications for green fluorescent protein (GFP) in the study of host-pathogen interactions. AB - The green fluorescent protein (GFP) from Aequorea victoria is a novel fluorescent marker that has potential use in the study of bacterial pathogenicity. To explore some of the potential applications of GFP to the study of host-parasite interactions, we constructed two GFP expression vectors suitable for different facultative intracellular bacterial pathogens. The first expression vector was tested in the enteric pathogens, Salmonella typhimurium and Yersinia pseudotuberculosis, and the second vector tested in Mycobacterium marinum (Mm). Both expression vectors were found to be stable and to direct high levels of GFP synthesis. Standard epifluorescence microscopy was used to detect all three bacterial pathogenic species during the early and late stages of infection of live mammalian cells. Mm expressing gfp was also visualized in infected animal tissues. gfp expression did not adversely affect bacterial survival, nor did it compromise entry into mammalian cells or their survival within macrophages. In addition, all three gfp-expressing bacterial pathogens could be detected and sorted in a flow cytometer, either alone or in association with epithelial cells or macrophages. Therefore, GFP not only provides a convenient tool to image pathogenic bacteria, but allows the quantitative measurement of bacterial association with mammalian cells. PMID- 8707056 TI - Chemistries and colors of bioluminescent reactions: a review. AB - Many different organisms, ranging from bacteria and fungi to fireflies and fish, are endowed with the ability to emit light, but the bioluminescent systems are not evolutionarily conserved: genes coding for the luciferase proteins (Lase) are not homologous, and the luciferins are also different, falling into many unrelated chemical classes. Biochemically, all known Lase are oxygenases that utilize molecular oxygen to oxidize a substrate (a luciferin; literally the "light-bearing' molecule), with formation of a product molecule in an electronically excited state. The color of the light may differ, even though the same luciferin/Lase system underlies the reaction. Filters or differences in Lase structure are responsible in some cases; in others a secondary emitter associated with a second protein is involved. In the coelenterates a green fluorescent protein, whose chromophore is derived from the primary amino-acid sequence, results in a red shift of the emission. In the bacteria accessory proteins causing either blue- or red-shifts have been isolated from different species; the chromophores are noncovalently bound. Although radiationless energy transfer has been implicated in the excitation of such accessory emitters, this may not be so in all cases. PMID- 8707057 TI - A transposon for green fluorescent protein transcriptional fusions: application for bacterial transport experiments. AB - The movement of bacteria through groundwater is a poorly understood process. Factors such as soil porosity and mineralogy, heterogeneity of soil particle size, and response of the bacteria to their environment contribute to the pattern of bacterial flow. The identification of transported bacteria is often a limiting factor in both laboratory and field transport experiments. Two bacterial strains were modified for use in bacterial transport experiments: a strain of Escherichia coli harboring the pGFP plasmid and a strain of Pseudomonas putida modified with a Tn5 derivative, Tn5GFP1. The Tn5GFP1 transposon incorporates the gene (gfp) encoding green fluorescent protein (GFP) and can be used to mutagenize Gram bacteria. Fluorescent colonies were suspended in phosphate-buffered saline (PBS) at a concentration of approx. 10(9) bacteria/ml. A 10-cm glass column packed with quartz sand (diameter range 177-250 microns) was equilibrated with PBS prior to the forced flow introduction of the bacteria. Collected fractions were analyzed and the bacteria quantitated using a fluorescence spectrometer. Results demonstrate that the bacteria can be accurately tracked using their fluorescence, and that the intensity of the signal can be used to determine a C/Co ratio for the transported bacteria. The data show a rapid breakthrough of the bacteria followed by a characteristic curve pattern. A lower limit of detection of 10(5) cells was estimated based on these experiments. The Tn5GFP1 transposon should become a valuable tool for labeling bacteria. PMID- 8707058 TI - Bacterial plasmid conjugation on semi-solid surfaces monitored with the green fluorescent protein (GFP) from Aequorea victoria as a marker. AB - Horizontal transfer of the TOL plasmid was examined in Pseudomonas putida (Pp) KT2442 micro-colonies on semi-solid agar surfaces. Horizontal gene transfer is usually studied in large populations where all information is based on average estimates of the transfer events in the entire population. We have used the green fluorescent protein (GFP) from the jellyfish Aequorea victoria as a plasmid marker, in combination with single-cell observations. This provided hitherto unknown details on the distribution of cells active in conjugation. In the present study, donor cells containing the gfp gene expressed from the bacteriophage T7 phi 10 promoter on the TOL plasmid, and recipient cells expressing the corresponding phage RNA polymerase allowed us to monitor the occurrence of ex-conjugants as green fluorescent cells upon illumination with blue light (470-490 nm). Further, the recipients were labeled with the luxAB genes to distinguish micro-colonies of donor cells from recipient cells. We conclude that conjugal plasmid transfer in Pp KT2442 cells on semi-solid surfaces occurs mainly during a short period of time after the initial contact of donors and recipients, indicating that spread of the TOL plasmid is limited in static, but viable cultures. PMID- 8707059 TI - Expression of the green fluorescent protein-encoding gene from a tobacco mosaic virus-based vector. AB - A cDNA fragment encompassing the Aequorea victoria green fluorescent protein encoding gene (gfp) was introduced into a genomic cDNA clone of tobacco mosaic virus (TMV). Infectious RNA transcripts produced in vitro were used to inoculate tobacco (Nicotiana benthamiana) leaves. After 1-2 days, bright green fluorescent areas could be visualized upon illumination with a long-wave ultraviolet (UV) light source. The virus was capable of infecting and expressing gfp in plant tissues both locally (in the inoculated leaf) and systemically (throughout the entire plant). Continued observation of inoculated plants indicated that systemic infection resulted from a combination of two forms of virus movement: a slow, cell-to-cell spread with concomitant virus replication and gfp expression, and a rapid, vascular-mediated form of transport without gfp expression. PMID- 8707060 TI - Plant virus movement protein dynamics probed with a GFP-protein fusion. AB - A genetic fusion between the gene encoding green fluorescent protein (GFP) from the jellyfish Aequorea victoria, with that of the Ob-tobamovirus movement protein (MP) resulted in the expression of a fluorescent fusion protein (MP::GFP) that was fully biologically active in mediating the cell-to-cell spread of the Ob virus. The MP::GFP fusion was used to follow in planta the subcellular trafficking of MP. GFP-tagged MP was transiently expressed and found to be associated with several subcellular compartments and structures including trans wall structures, presumably plasmodesmata, and filament structures. The MP::GFP fusion can be used to monitor MP association with host proteins and structures, and for the isolation of interacting host components. PMID- 8707061 TI - Green fluorescent protein and its derivatives as versatile markers for gene expression in living Drosophila melanogaster, plant and mammalian cells. AB - We have investigated the utility of the green fluorescent protein (GFP) as a marker for gene expression in living adult Drosophila melanogaster (Dm) and cultured plant and mammalian cells. Using Dm, we generated transgenic flies bearing a glass-responsive gfp fusion gene to test the utility of GFP as a spatial reporter. In the adult living fly, GFP is clearly visible in the ocelli and the eye. We have optimized the use of filters for distinguishing the GFP signal from abundant autofluorescence in living Dm. In addition, we have used GFP to identify photoreceptor cells in pupal eye cultures that have been fixed and stained according to standard histological procedures. GFP was also detected in individual living plant cells following transient transfection of soybean suspension cultures, demonstrating that GFP is an effective transformation marker in plant cells. Similarly, transient transfection of mammalian cells with a modified form of GFP, S65T, allowed detection of single living cells expressing the reporter. This modified form of GFP gave a robust signal that was resistant to photobleaching. We then used a CellScan system exhaustive photon reassignment (EPR) deconvolution algorithm to generate high-resolution three-dimensional images of GFP fluorescence in the living cell. PMID- 8707062 TI - Green fluorescent protein marks skeletal muscle in murine cell lines and zebrafish. AB - The green fluorescent protein (GFP) acts as a vital dye upon the absorption of blue light. When the gfp gene is expressed in bacteria, flies or nematodes, green fluorescence can be directly observed in the living organism. We inserted the cDNA encoding this 238-amino-acid (aa) jellyfish protein into an expression vector containing the rat myosin light-chain enhancer (MLC-GFP) to evaluate its ability to serve as a muscle-specific marker. Transiently, as well as stably, transfected C2C12 cell lines produced high levels of GFP distributed homogeneously throughout the cytoplasm and was not toxic through several cell passages. Expression of MLC-GFP was strictly muscle-specific, since Cos 7 fibroblasts transfected with MLC-GFP did not fluoresce. When GFP and beta Gal markers were compared, the GFP signal was visible in the cytoplasm of the living cell, whereas visualization of beta Gal required fixation and resulted in deformation of the cells. When the MLC-GFP construct was injected into zebrafish embryos, muscle-specific gfp expression was apparent within 24 h of development. gfp expression was never observed in non-muscle tissues using the MLC-GFP construct. Transgenic fish continued to express high levels of gfp in skeletal muscle at 1.5 months, demonstrating that GFP is an effective marker of muscle cells in vivo. PMID- 8707063 TI - Requirements for green fluorescent protein detection in transgenic zebrafish embryos. AB - We have generated transgenic (Tg) lines of zebrafish in which the green fluorescent protein (GFP)-encoding gfp cDNA is driven by the Xenopus laevis ef1 alpha enhancer/promoter; Tg embryos from most of these lines show detectable fluorescence throughout their body. We have investigated the copy number of the Tg genes in fluorescent and non-fluorescent lines, in order to determine how this affects the production of detectable levels of GFP in the zebrafish embryo. Additionally, we have injected purified recombinant GFP into embryos to determine the intracellular GFP concentration required for detection, both when all of the cells in the embryo contain GFP and when only a few do. PMID- 8707065 TI - Doctor Net: Physicians can create their own free home page on the Web. PMID- 8707064 TI - Paget's disease of bone: why not use less expensive skull films? PMID- 8707066 TI - A 'revolutionary class' of scapegoats. PMID- 8707067 TI - Diffuse female hair thinning. PMID- 8707068 TI - Kidney protection: how to prevent or delay chronic renal failure. AB - In patients at risk for chronic renal failure, early clinical interventions have been shown to prevent or delay progression of the disease. A low-protein diet is recommended when the serum creatinine is between 1.5 and 2.5 mg/dl. Benefit from protein restriction can only be achieved if energy intake is greater than 35 kcal/kg/d. To preserve skeletal muscle, correct metabolic acidosis with oral sodium bicarbonate supplementation. For hypertension control and renal protection, start ACE inhibitors when serum creatinine is less than 2 mg/dl. Correct anemia with erythropoietin to improve or maintain quality of life when the hematocrit falls below 30%. For the diabetic, ACE inhibitors and glycemic control have been shown to show the rate of renal failure. PMID- 8707069 TI - Osteoarthritis: what to look for, and when to treat it. Interview by Peter Pompei. AB - In patients with suspected osteoarthritis, start with a thorough history aimed at finding the mechanical component of the condition. Ask about past and current activities that may be loading or stressing a painful joint. Conditions to consider in the differential diagnosis are those affecting soft tissues-muscles, tendons, ligaments, and bursae. Injecting the tender are with lidocaine will help identify a separate or coexisting soft tissue disorder. Before considering drug therapy, attempt to correct any mechanical factors that may be contributing to the problem. Modify the patient's exercise routine, if necessary, and begin patients on a walking aid if a weight-bearing joint is affected. PMID- 8707070 TI - Type II diabetes: a treatment approach for the older patient. AB - Noninsulin-dependent (type II) diabetes mellitus occurs most commonly with advancing age. The disease is accompanied by significant risk for morbidity of many organ systems and by risk for premature death. Morbidity and mortality are linked to persistent elevation of blood glucose concentration of even moderate degree. Through diet, weight reduction, exercise, and drug therapies, it is possible to bring blood glucose concentrations to close to normal levels and to reduce the likelihood of development or progression of diabetic complications. PMID- 8707071 TI - Geriatrics photo quiz. Dupuytren's contracture. PMID- 8707072 TI - Ultrasound for gastroenterologists. PMID- 8707073 TI - Natural history of reflux oesophagitis: a 10 year follow up of its effect on patient symptomatology and quality of life. AB - BACKGROUND: Although oesophagitis is the most common diagnosis made at upper gastrointestinal endoscopy, data on the longterm outcome of affected patients are sparse. AIMS: This study assessed the level of reflux symptoms, quality of life, drug consumption, and complications in patients at least 10 years after diagnosis of oesophagitis at one centre. PATIENTS: One hundred and fifty two patients with typical reflux symptoms and a first time diagnosis by endoscopy of grade I-III oesophagitis between 1981 and 1984, were followed up using a postal questionnaire and telephone interview. RESULTS: Eighteen of 152 patients had died, 33 failed to respond, and 101 replied (mean follow up 11 years, range 121-160 months). Over 70% of patients still had heartburn at least daily (32%) or weekly (19%) or required daily acid suppression treatment (20%). Two patients (2%) had developed oesophageal strictures and one had Barrett's oesophagus. Two of eight quality of life scores (physical function and social function) measured by the Short Form-36 were significantly lower than Northern Ireland population scores. CONCLUSION: Nearly three quarters of patients previously diagnosed as having oesophagitis still had significant morbidity related to gastro-oesophageal reflux disease more than 10 years after diagnosis. Some quality of life scores were significantly lower than those of the general population. PMID- 8707074 TI - Laparoscopic Nissen fundoplication--200 consecutive cases. AB - PATIENTS: Laparoscopic Nissen fundoplication was undertaken in 200 patients between 1991 and 1994. METHODS: Pre-operative assessment included symptom score, endoscopy, manometry, and 24 hour pH monitoring of the oesophagus. Patients were evaluated at three and 12 months after surgery with symptom scoring and 96 patients also underwent 24 hour pH studies at three to six months postoperatively. RESULTS: In the first 100 patients median duration of operation was 155 minutes (range: 70-330), conversion rate to laparotomy was 7%, median hospital stay was three days (range: 2-57), and total morbidity was 16%. This compared with a median operation time of 120 minutes (60-240) (p = 0.0003, 95% CI 10, 40), a conversion rate of 2% (p = 0.2), a hospital stay of three days (1-18) (p = 0.0016, 95% CI 0, 1), and total morbidity of 7% (p = 0.15) in the second 100 patients. Median total symptom scores fell from 5/9 to 0/9 after fundoplication (< 0.0001) while median 24 hour oesophageal acid exposure in 96 patients was reduced from 10% to 1% (p < 0.001). CONCLUSIONS: Laparoscopic Nissen fundoplication is a safe and effective procedure for gastro-oesophageal reflux disease. With experience, the duration of operation falls and the hospital stay is shorter. Shortterm symptomatic and pH results are consistently improved by surgery. PMID- 8707075 TI - Effect of acute stress on oesophageal motility in patients with gastro oesophageal reflux disease. AB - BACKGROUND AND AIM: Sixty four per cent of people with heartburn believe that it is exacerbated by stress. An alteration in oesophageal motility is one possible mechanism for this apparent change with stress. This study aimed to assess the effect of acute stressors on oesophageal motility in patients with gastro oesophageal reflux disease (GORD). METHODS: Sixty patients were studied. Twenty had oesophagitis, 20 had increased oesophageal acid exposure on pH monitoring but no endoscopic oesophagitis, and 20 had neither oesophagitis nor abnormal oesophageal acid exposure. Oesophageal motility was studied in these patients during psychological stress (Stroop test) and physical stress (cold pressor test). RESULTS: Blood pressure (BP) and heart rate increased in response to both stressors (mean systolic BP increased by > 10 mm Hg, diastolic BP by > 4 mm Hg and heart rate by > 3 beats per minute (p < 0.00001). The amplitude, duration, and velocity of propagation of oesophageal peristaltic contractions were not altered by the stressors. The percentage of simultaneous waves increased in patients with oesophagitis during the cold pressor test (median increase in these patients was 6% (p < 0.05)). This effect was not noted in the patients without oesophagitis. CONCLUSION: Acute stressors did not induce significant changes in oesophageal motility in patients with GORD but no oesophagitis. For these patients, dysmotility is not likely to be a cause of oesophageal symptoms which are exacerbated by stress. There was, however, a significant increase in simultaneous waves during cold pressor stress in patients with oesophagitis. PMID- 8707076 TI - Induction of gastric epithelial apoptosis by Helicobacter pylori. AB - BACKGROUND: Helicobacter pylori may promote gastric carcinogenesis through increasing gastric epithelial cell proliferation. How H pylori does so is unknown. Programmed, non-necrotic, cell death (apoptosis) occurs throughout the gut and is linked to proliferation. It was hypothesised that H pylori may induce hyper-proliferation through increasing apoptosis. AIM: To measure the effect of H pylori infection on gastric epithelial apoptosis in situ. PATIENTS: Patients with duodenal ulcers treated to eradicate H pylori and patients with H pylori negative non-ulcer dyspepsia. METHODS: Retrospective quantification of apoptotic epithelial cells in situ from formalin fixed biopsy specimens, counted after staining by terminal uridine deoxynucleotidyl nick end-labelling. RESULTS: In the uninfected stomach, apoptotic cells were rare and situated in the most superficial portion of gastric glands (mean 2.9% of epithelial cells). In H pylori infection, they were more numerous and were located throughout the depth of gastric glands, comprising 16.8% of epithelial cells, falling to 3.1% after H pylori eradication, p = 0.017. Apoptotic cell number did not correlate with the degree of histological gastritis. CONCLUSIONS: These results suggest that H pylori induces epithelial apoptosis in vivo. Increased apoptosis may be the stimulus for a compensatory hyperproliferative and potentially preneoplastic response in chronic H pylori infection. PMID- 8707077 TI - Gastric mucosal superoxide dismutases in Helicobacter pylori infection. AB - BACKGROUND: The mucosal pathology of Helicobacter pylori infection may in part be due to excessive production of reactive oxygen metabolites (ROMs) by phagocytes. The influence of H pylori infection on mucosal superoxide dismutases, some major scavenger enzymes of ROM was investigated. In humans superoxidase dismutase is present in at least two forms-that is, mitochondrial manganese (Mn)-superoxide dismutase and cytoplasmic copper-zinc (CuZn)-superoxide dismutase. METHODS: The amount and activity of both superoxide dismutases were measured, respectively by enzyme linked immunosorbent assay (ELISA) and spectrophotometrical enzyme activity assay, in gastric biopsy homogenates of patients with normal mucosa (n = 39) and in patients with H pylori related gastritis (n = 71). Infection and gastritis were confirmed by a combination of culture, serology, and histology. RESULTS: The amount (p < 0.001) and activity (p < or = 0.05) of Mn-superoxide dismutase were increased by about twofold to three-fold, whereas the amount and activity of CuZn-superoxide dismutase showed a slight decrease in gastric mucosa of patients with H pylori gastritis, in both antrum and corpus, compared with normal mucosa of patients without H pylori infection. Mn-superoxide dismutase concentrations in biopsy specimens of histologically normal corpus from patients with an inflamed antrum were significantly higher (p < 0.01) than that of patients with a histologically normal antrum. CONCLUSION: H pylori infection has a differential effect on mitochondrial and cytoplasmic superoxide dismutase in the gastric mucosa, reflected by a pronounced increase in the cytokine inducible Mn-superoxide dismutase and a marginal decrease in the constitutive CuZn superoxide dismutase. PMID- 8707078 TI - Expression of CD44 and its variants on gastric epithelial cells of patients with Helicobacter pylori colonisation. AB - BACKGROUND: Studies have suggested that expression of the adhesion molecule CD44 may be of prognostic importance in gastric cancer. In addition, there is strong evidence that Helicobacter pylori has a role in gastric cancer. AIMS: To determine the expression of CD44 and its variants (v6, v9) and HLA class II molecules on human gastric epithelial cell and intraepithelial lymphocytes in patients with and without H pylori infection. PATIENTS: Eighteen patients (seven men and 11 women) attending for endoscopic evaluation because of upper gastrointestinal symptoms were included. An additional 10 patients (five men and five women) were analysed for CD44 variant expression). METHODS: Biopsy specimens were taken from the gastric antrum during endoscopy. Gastric epithelial cells and intraepithelial lymphocytes were examined by two colour flow cytometry and compared in patients with and without H pylori infection. RESULTS: Expression of CD44 and its variants (CD44 v9) was increased in epithelial cells but not in intraepithelial lymphocytes. Both epithelial cells and intraepithelial lymphocytes expressed higher levels of HLA class II molecules (DR and DP), possibly as a result of local cytokine production. Furthermore, results showed upregulation of CD44 on a gastric epithelial cell line (AGS) by cytokines and peripheral blood mononuclear cell supernatant. CONCLUSIONS: These data suggest that H pylori, either directly or through a local inflammatory response, is responsible for increased expression of CD44 and its variant CD44 v9. These data are of potential importance in relation to increased expression of CD44 and CD44 v9 on gastric carcinoma. PMID- 8707079 TI - Gastric metaplasia and duodenal ulcer disease in children infected by Helicobacter pylori. AB - BACKGROUND: Helicobacter pylori infection of the gastric mucosa is vital in the pathogenesis of duodenal ulcer disease. H pylori will only colonise gastric epithelium and its association with duodenal disease is therefore not easily explained. AIMS: To determine if gastric metaplasia in the duodenum increases the risk of duodenal ulcer disease in children infected with H pylori. PATIENTS: All children undergoing upper endoscopy over a 20 month period in a children's hospital in Ireland. METHODS: Two biopsy specimens were obtained from the antral mucosa and two from the first part of the duodenum. One antral biopsy specimen was used in a rapid urease test (Clo Test). Biopsy sections were stained with haematoxylin and eosin and also with cresyl violet for identification of H pylori. Periodic acid Schiff (PAS) stain was performed to identify areas of gastric metaplasia. RESULTS: Gastric and duodenal biopsy specimens were obtained from 148 patients (M:F 1:2:1). Twenty five children (17%) had H pylori positive gastritis. Thirty four children (23%) had gastric metaplasia in the duodenum. Nine per cent of children under the age of 8 years had gastric metaplasia compared with 38% in those 12 years of age or over (p < 0.005). Seven children had duodenal ulcer disease. Gastric metaplasia was present in six of seven (86%) children with duodenal ulcer disease compared with 28 of 141 (20%) without ulceration (p < 0.001). While both H pylori and gastric metaplasia were each significant risk factors for duodenal ulcer disease, the combined presence of both factors was associated with a pronounced increase in duodenal ulcer disease. Duodenal ulcer disease occurred in over 50% of children with both H pylori infection and gastric metaplasia. In contrast duodenal disease did not occur in children (0 of 100) when both were absent. CONCLUSION: The presence of gastric metaplasia in the duodenum is the major risk factor for duodenal ulcer disease in patients colonised by H pylori. PMID- 8707081 TI - Relation of number of positive lymph nodes to the prognosis of patients with primary gastric adenocarcinoma. AB - BACKGROUND: No nodal grouping category of gastric cancer has been universally accepted for the grading of the effectiveness of therapeutic regimens. AIMS: To establish an appropriate nodal grouping as a forecaster of distant disease and test its validity as a determinant in survival. PATIENTS: Five hundred and ten patients who underwent curative resections for gastric cancer were studied. METHODS: Retrospectively analyse the prognostic significance of the number of metastatic lymph nodes. RESULTS: A total of 17 176 lymph nodes with an average of 34 per specimen were removed, of which 2811 (16%) showed metastases. Among the 510 patients, 287 (56%) had lymph node metastases, with an average of 9.8 per metastatic case. The survival of all patients was related to their nodal status, an abrupt decrease in survival was seen between 0 and 1 and 4 compared with 5 or more modes while little difference in survival existed among 1, 2, 3, and 4, and among 5, 6, 7, and 8 positive nodes. Multivariate analysis showed that the number of positive nodes (1-4, 5-8 versus > or = 9; relative risk 2.2) and depth of cancer invasion (three levels; relative risk 1.9) were independently correlated with survival. The current nodal stage was not a prognostic factor. CONCLUSIONS: Gastric cancer patients with 0, 1 to 4, 5 to 8, and > 9 positive nodes may represent four appropriate prognostic groups and should be adopted for classification of nodal stage in gastric cancer. PMID- 8707080 TI - Effect of allopurinol, sulphasalazine, and vitamin C on aspirin induced gastroduodenal injury in human volunteers. AB - BACKGROUND: The mechanisms of aspirin induced gastroduodenal injury are not fully understood. Aspirin induces the release of reactive oxygen metabolites in animal models, which may contribute to mucosal injury. AIMS: To investigate the effects of aspirin administered with placebo or antioxidants on gastric mucosal reactive oxygen metabolite release and gastroduodenal injury in human volunteers. SUBJECTS: Fourteen healthy volunteers participated in the study (seven male; mean age 27 years, range 20-40). METHODS: In a double blind, randomised, crossover study, volunteers received aspirin 900 mg twice daily and either placebo, allopurinol 100 mg twice daily, sulphasalazine 1 g twice daily or vitamin C 1 g twice daily for three days. Injury was assessed endoscopically and by quantifying mucosal reactive oxygen metabolite release by measuring chemiluminescence before and after each treatment. The effect on prostanoids was determined by measuring ex vivo antral prostaglandin E2 (PGE2) synthesis and serum thromboxane B2 (TXB2). RESULTS: No drug reduced any parameter of gastric injury but vitamin C reduced duodenal injury assessed by Lanza score (p < 0.005). Chemiluminescence increased after aspirin both with placebo (p < 0.05) and vitamin C (p < 0.05). Post treatment chemiluminescence was lower in subjects taking allopurinol (p < 0.05) or sulphasalazine (p < 0.005) than in those taking placebo with aspirin. CONCLUSIONS: In this study, aspirin induced gastric injury was associated with reactive oxygen metabolite release. This was reduced by sulphasalazine and allopurinol, although macroscopic injury was not affected. Vitamin C, however, was shown to have a previously unrecognised protective effect against aspirin induced duodenal injury. PMID- 8707082 TI - Malignancy and survival in dermatitis herpetiformis: a comparison with coeliac disease. AB - BACKGROUND: Dermatitis herpetiformis is a lifelong, gluten sensitive skin disease. Patients with dermatitis herpetiformis, similar to patients with coeliac disease not adhering to a gluten free diet, seem to have increased risk for lymphoma. AIMS: This study looked at the occurrence of malignancy and survival of patients with dermatitis herpetiformis and compared the results with those seen in patients with coeliac disease or in the general population. PATIENTS: A total of 305 adult patients with dermatitis herpetiformis diagnosed at the University Hospital of Tampere in 1970-1992 were studied. Most patients started a gluten free diet and at the end of the study 93% of the patients were adhering to the diet. A control group comprised 383 adult patients with coeliac disease, 81% of them adhered to a gluten free diet, 6% had a normal diet, and in 13% the diet history remained unknown. METHODS: The occurrence of malignant diseases and survival of the patients were assessed up to the end of 1993. Standardised incidence ratios (SIR) with 95% confidence intervals were used for the malignant diseases. The survival of the patients was compared with that of the general population. RESULTS: Thirteen (4.3%) patients with dermatitis herpetiformis developed 14 malignant disorders during the follow up (SIR 1.25; 95% confidence intervals 0.68 to 2.09). A non-Hodgkin's lymphoma occurred in four patients with dermatitis herpetiformis, significantly more than expected (SIR 10.3; 2.8-26.3). Thirteen (4.3%) patients with dermatitis herpetiformis died during the follow up but there was no increased general mortality. In coeliac disease, 13 (3.4%) patients developed malignancy (SIR 1.16; 0.62 to 1.97), 31 (8.1%) patients died but the survival rate did not differ from that in the general population. CONCLUSIONS: The incidence of non-Hodgkin's lymphoma was significantly increased in patients with dermatitis herpetiformis. The results also confirm that the patients with dermatitis herpetiformis treated mainly with a gluten free diet have no increased general mortality. PMID- 8707083 TI - Comparative effects of dietary nucleoside-nucleotide mixture and its components on endotoxin induced bacterial translocation and small intestinal injury in protein deficient mice. AB - BACKGROUND: Nucleoside-nucleotide mixture has been shown to improve gut morphology and reduce the incidence of bacterial translocation in protein deficient mice. AIMS: To compare the reparative effect of nucleoside-nucleotide mixture and their individual components on maintenance of gut integrity and bacterial translocation based on their differential metabolism and utilisation. METHODS: ICR (CD-1) mice were randomised into eight groups of 10 animals each and fed 20% casein diet (control), protein free diet, or protein free diet supplemented with 3 M cytidine, uridine, thymidine, inosine, guanosine monophosphate, or nucleoside-nucleotide mixture for four weeks. On the fourth week, each mouse was injected lipopolysaccharide intraperitoneally (50 micrograms/500 microliters) and the incidence of bacterial translocation, caecal bacterial populations, and the ileal histology, noted 48 hours later. RESULTS: The death rate in the control group was 40% compared with 10% in the nucleoside nucleotide mixture and 20% each in the individual components groups, respectively. Bacterial translocation to the mesenteric lymph node did occur in 100% of the surviving mice fed the control diet in comparison with 44% (nucleoside-nucleotide), 50% (cytidine), 75% (thymidine), 75% (uridine), 63% (inosine), and 63% (guanosine monophosphate). Histologically, the damage to the gut was more distinct in the protein free diet group. Villous height, crypt depth, and wall thickness in the nucleoside-nucleotide mixture group mean (SEM) (5.01 (0.34); 0.87 (0.14); 0.33 (0.10)), were respectively, higher compared with the protein free diet (3.34 (0.34); 0.61 (0.03); 0.18 (0.04)) group. In the cytidine group, crypt depth (0.86) (0.08)), and wall thickness (0.30 (0.002)) were higher. The same measurements in the components groups tended to be higher than the protein free diet group. Caecal bacterial populations were, however, similar in all groups. CONCLUSIONS: These results suggest that dietary nucleosides and nucleotides are essential nutrients for intestinal repair; nucleotides or cytidine provide a better response. PMID- 8707084 TI - HLA class II gene frequencies in Crohn's disease: a population based analysis in Germany. AB - BACKGROUND: Ulcerative colitis is the only known inflammatory bowel disease associated with particular HLA alleles. Whereas the association with the HLA DRB1*15 allele has been described in several independent studies for ulcerative colitis, no contribution of HLA alleles to susceptibility in Crohn's disease has yet been shown. AIM: This study was designed to study the strength of association of HLA class II alleles as risk markers for Crohn's disease in a homogenous population in Germany. PATIENTS: A total of 4251 randomly selected control subjects, and 162 unrelated subjects with Crohn's disease were studied. Subjects were studied for their HLA-DRB1, HLA-DQA1, and HLA-DQB1 alleles. METHOD: HLA DNA typing was performed after locus specific amplification with the polymerase chain reaction and reverse dot blot hybridisation. RESULTS: The HLA-DRB1*07 was the only HLA class II allele found to be significantly associated with Crohn's disease (relative risk (RR) = 1.9, 95% CI: 1.66 to 2.14; p = 0.0001). This association remained significant after correction for the number of DRB1 alleles compared. In patients with disease onset before 35 years the RR for the disease in HLA-DRB1*07 positive subjects was found to be higher (RR = 3.1, 95% CI: 2.44 to 3.76). The HLA-DRB1*03 was significantly decreased in frequency in Crohn's disease (RR = 0.5, 95% CI: 0.39 to 0.61; p = 0.0028). CONCLUSION: The HLA-DRB1*07 allele provides risk for the disease especially in patients with younger ages of onset. These data also provide indirect evidence for an immunogenetically based heterogeneity of the disease. PMID- 8707085 TI - Supplementary enteral nutrition maintains remission in paediatric Crohn's disease. AB - BACKGROUND: Liquid diets given enterally combined with "bowel rest' are efficacious in the treatment of active Crohn's disease, but rapid recrudescence of gastrointestinal symptoms after resumption of a normal diet is common. AIMS: This study examined whether continuation of enteral nutrition as a nocturnal supplement to an ad libitum daytime intake of a normal diet increased the length of remission of Crohn's disease in children. PATIENTS AND METHODS: Children and adolescents with active Crohn's disease treated successfully with exclusive enteral nutrition were classified retrospectively according to whether they continued supplementary enteral nutrition or not. Time to relapse and linear growth were compared between the two cohorts. RESULTS: Between January 1986 and December 1992, 65 patients aged 7-17 years (mean (SD) 13.6 (2.1) years) (36 males, 29 females) with Crohn's disease in exacerbation were treated for > or = four weeks by bowel rest and nasogastric tube feeding of an oligopeptide or amino acid based formula. At first follow up visit, remission (fall in Paediatric Crohn's Disease Activity Index, PCDAI to < or = 20) was achieved in 47 of 65 (72%) patients. Subsequently, 20 of these 47 (43%) relapsed by six months and 28 of 47 (60%) by 12 months. Patients who continued nasogastric supplementary feeding (n = 28) after resumption of an otherwise normal diet remained well longer than those who discontinued nocturnal supplements completely (n = 19) (p < 0.02). Furthermore, continued use of nasogastric supplements before completion of puberty was associated with improved linear growth. CONCLUSION: After successful treatment of active Crohn's disease by exclusive enteral nutrition, supplementary enteral nutrition without restriction of normal diet is associated with prolongation of remission and improved linear growth in children and adolescents. PMID- 8707086 TI - Leucocyte typing, cytokine expression, and epithelial turnover in the ileal pouch in patients with ulcerative colitis and familial adenomatous polyposis. AB - BACKGROUND/AIMS: Conventional histopathology, leucocyte typing, cytokine mRNA expression, and crypt cell turnover were compared in ileal pouch biopsy specimens from patients with ulcerative colitis (UC) and familial adenomatous polyposis (FAP). METHODS: Biopsy specimens were taken from 17 patients with UC and seven with FAP at a median interval of 19 months (range 2-120) after ileostomy closure. All contained both epithelium and lamina propria. Cryostat sections were stained for lymphocyte subtypes (CD3, CD4, CD8), macrophages (CD68), common leucocyte antigen (CD45), and Ki-67, using a three stage immunoperoxidase reaction. Cytokine mRNA expression for interleukins 2 and 6, tumour necrosis factor alpha, and interferon gamma was studied using an in situ hybridisation technique. RESULTS: Lymphocyte subtype and macrophage populations in epithelium and lamina propria were similar in UC and FAP. The labelling index (Ki-67) was significantly increased in biopsy specimens from patients with UC (UC median = 43.3 (interquartile range (IQR) 38.9-48.2) v FAP 34.9 (29.9-35.2), p < 0.05). There was little or no epithelial mRNA expression for any cytokine in any of the specimens. Lamina propria mRNA expression for interleukin 2 was significantly increased in UC (UC median (IQR) 10.7 (5.4-14.2) cells per unit area v FAP 2.8 (1.5-6.6) p < 0.05) but not for tumour necrosis factor alpha, interleukin 6, and interferon gamma. CONCLUSIONS: While static morphological assessment (leucocyte type, conventional histopathological examination) was similar, tests of cell function (mRNA expression and labelling index) were different in ileal pouches in patients with UC compared with FAP. The study also showed that mRNA expression occurred almost entirely in the lamina propria. PMID- 8707087 TI - MUC2 is the prominent colonic mucin expressed in ulcerative colitis. AB - BACKGROUND: It has been shown that MUC2 is the prominent mucin synthesised in healthy colon. AIM: To identify the predominant mucins in ulcerative colitis (UC) and to study their biosynthesis. METHODS AND RESULTS: Mucin was purified from UC resection specimens. This mucin on sodium dodecylsulphate polyacrylamide gel electrophoresis (SDS-PAGE) presented as one, high molecular weight, periodic acid/Schiff's reagent (PAS) stainable band. Amino acid composition showed a close resemblance to that of MUC2. Immunoprecipitation with a specific anti-MUC2 antiserum confirmed that this mucin was MUC2. In addition, on the mRNA level MUC2 was also the most prominent mucin expressed in UC. Polyclonal antiserum was elicited, mainly recognising mucin peptide epitopes of UC and normal colonic mucin. Biosynthetic studies with [35S]amino acids showed that the MUC2-precursor in UC displayed a molecular mass on SDS-PAGE of approximately 600 kDa. This precursor was converted into a mature MUC2 with anomalous mobility on SDS-PAGE of 550 kDa and was secreted. Only this 550 kDa band could be labelled with [35S]sulphate and stained by PAS. CONCLUSIONS: This study shows that in parallel with the mucin expression in healthy controls, MUC2 is the major mucin expressed in UC. Qualitatively, MUC2 biosynthesis seems unchanged in UC. PMID- 8707088 TI - Soluble Fc gamma receptor III (CD 16) and eicosanoid concentrations in gut lavage fluid from patients with inflammatory bowel disease: reflection of mucosal inflammation. AB - BACKGROUND: Activated neutrophils cause tissue injury in inflammatory bowel disease (IBD). Upon activation, they shed soluble Fc gamma IIIb receptors (sFc gamma RIIIb). The subsequent inflammatory response is modulated by several mediators, including neutrophil derived leukotriene B4 (LTB4), thromboxane B2 (TXB2), and prostaglandin E2 (PGE2). The aim of this study was to determine the value of gut lavage sFc gamma RIII and eicosanoid measurements for the assessment of mucosal inflammation in IBD. METHODS: A total of 18 patients with active IBD, 10 ulcerative colitis (UC), and eight Crohn's disease (CD), and 12 control patients underwent whole gut lavage. Disease activity, endoscopic appearance, and histopathology were graded. Samples were processed for the determination of sFc gamma RIIIb, LTB4, PGE2, and TXB2. RESULTS: Soluble Fc gamma RIIIb concentrations were increased in both IBD groups. Significant correlations were seen between sFc gamma RIIIb and LTB4 values with histology scores. Mean eicosanoid lavage fluid concentrations in control patients were 14.1 pg/ml for LTB4, 5.6 pg/ml for PGE2, and 397 pg/ml for TXB2. Concentrations of all eicosanoids in IBD patients were significantly increased: LTB4 in UC: mean 73.2 pg/ml, in CD: 96.4 pg/ml (both p < 0.01 v controls). PGE2 in UC: 20.2 pg/ml, in CD: 43.4 pg/ml (p < 0.01). TXB2 in UC: 719.3 pg/ml, in CD: 180.6 pg/ml (both p < 0.05). CONCLUSIONS: Whole gut lavage fluid analysis is an effective method to study mucosal eicosanoid production. Soluble Fc gamma RIIIb concentrations in gut lavage fluid closely correlate with histological signs of mucosal inflammation and with lavage LTB4 concentration. These data suggest that lavage Fc gamma RIIIb assessment may be used as a simple assay to estimate mucosal neutrophil infiltration in IBD. PMID- 8707089 TI - Butyrate enema therapy stimulates mucosal repair in experimental colitis in the rat. AB - BACKGROUND: The short chain fatty acid (SCFA) butyrate provides energy for colonocytes, stimulates colonic fluid and electrolyte absorption and is recognised as an effective treatment for multiple types of colitis. AIM: To examine the impact of butyrate enema therapy on the clinical course, severity of inflammation, and SCFA stimulated Na+ absorption in a chronic experimental colitis. METHODS: Distal colitis was induced in rats with a trinitrobenzenesulphonic acid (TNBS) enema. Five days after induction, rats were divided into groups to receive: no treatment, saline enemas, or 100 mM Na butyrate enemas daily. On day 24, colonic damage score and tissue myeloperoxidase (MPO) activity were evaluated. Colon was mounted in Ussing chambers and Na+ transport and electrical activities were measured during a basal period and after stimulation with 25 mM butyrate. RESULTS: In the untreated and the saline enema treated TNBS groups, diarrhoea and extensive colonic damage were seen, associated with increased tissue MPO activities and absent butyrate stimulated Na+ absorption. In contrast, in the butyrate enema treated TNBS group, diarrhoea ceased, colonic damage score improved, and tissue MPO activity as well as butyrate stimulated Na+ absorption recovered to control values. CONCLUSION: Butyrate enema therapy stimulated colonic repair, as evidenced by clinical recovery, decreased inflammation, and restoration of SCFA stimulated electrolyte absorption. PMID- 8707090 TI - Outcome of restorative proctocolectomy with ileal reservoir for ulcerative colitis: comparison of distal colitis with more proximal disease. AB - BACKGROUND: An increasing number of patients with severe or refractory ulcerative colitis involving only the rectum and sigmoid colon are being offered restorative proctocolectomy with ileal reservoir but very few data are available concerning the outcome for these patients. AIM: This study was designed to compare the outcome of ileal pouch procedures for distal ulcerative colitis with procedures performed for more extensive disease. PATIENTS: A consecutive series of 177 patients undergoing restorative proctocolectomy for ulcerative colitis between January 1984 and December 1994. METHODS: Data were collected prospectively in a dedicated ileal pouch database and included demographic details, indication for surgery, surgical procedures performed, early (< 30 days) and late morbidity, functional outcome, and histopathology. RESULTS: There was no mortality in the series. The incidence and range of early morbidity (< 30 days) and the functional outcome (daytime stool frequency, nocturnal frequency, and the incidence of incontinence) were similar for all groups. Log rank analysis of Kaplan-Meier estimates showed no significant difference between groups in the likelihood of developing pouchitis (p > 0.2). CONCLUSIONS: Patients undergoing restorative proctocolectomy for distal colitis experience a similar outcome to patients with more extensive disease. These data refute the hypothesis that pouchitis is more common in patients with total colitis. PMID- 8707091 TI - Sulindac induced regression of colorectal adenomas in familial adenomatous polyposis: evaluation of predictive factors. AB - BACKGROUND: Sulindac, a non-steroidal anti-inflammatory drug, causes regression of colorectal adenomas in patients with familial adenomatous polyposis (FAP) but the response is variable. Specific clinical factors predictive of sulindac induced regression have not been studied. METHODS: 22 patients with FAP were given sulindac 150 mg orally twice a day. Polyp number and size were determined before treatment and at three months. The relation of nine clinical factors to polyp regression (per cent of baseline polyp number after treatment) was evaluated by univariate and multivariate analysis. RESULTS: After three months of sulindac, polyp number had decreased to 45 per cent of baseline and polyp size to 50 per cent of baseline (p < 0.001 and p < 0.01, respectively). Univariate analysis showed greater polyp regression in older patients (p = 0.004), those with previous colectomy and ileorectal anastomosis (p = 0.001), and patients without identifiable mutation of the APC gene responsible for FAP (p = 0.05). With multivariate regression analysis, response to sulindac treatment was associated with previous subtotal colectomy. CONCLUSIONS: Sulindac treatment seems effective in producing regression of colorectal adenomas of FAP patients with previous subtotal colectomy regardless of baseline polyp number and size. Changed sulindac metabolism, reduced area of the target mucosa, or changed epithelial characteristics after ileorectal anastomosis may explain these findings. PMID- 8707092 TI - Effects of longterm epidermal growth factor treatment on the normal rat colon. AB - BACKGROUND: Epidermal growth factor (EGF) exerts trophic effects on the mucosa of damaged and defunctioned colon, but the effects on the normal large bowel wall are not known. AIMS: To investigate the effect of systemic EGF treatment on growth and morphology of normal rat colon. METHODS: Rats were treated with subcutaneous biosynthetic EGF injections of 150 micrograms/kg/day for 28 days. The weight of the histological colonic wall layers and the luminal surface area were measured using quantitative morphometric analysis (stereology). The colon was subdivided into proximal and distal parts. RESULTS: EGF treatment increased the total colon wet weight by 23% compared with controls (p < 0.005). The weight increase occurred in the mucosal (33%) and the submucosal layers of the bowel wall (36%) and there was a 69% increase of the total luminal surface area (p = 0.001). In the proximal part of colon of EGF rats there was a 68% increase in mucosal weight (p < 0.005) accompanied by a 79% increase in the mucosal surface area compared with controls (p < 0.005), whereas submucosal and muscularis propria weights were identical. In distal colon, the mucosal weight increased 28% in the EGF group (p < 0.005), the mucosal surface area increased by 72% after treatment (p < 0.01). Furthermore there was a 34% increase in the weight of submucosa (p < 0.001) in the distal colon among EGF rats. CONCLUSIONS: Treatment of rats with EGF has a stimulating role on the mucosa and luminal surface area of the entire functioning colon and a trophic effect on the submucosa of the distal colon. PMID- 8707093 TI - Pathology of the rectal wall in solitary rectal ulcer syndrome and complete rectal prolapse. AB - BACKGROUND: The aetiology and pathology of rectal prolapse and solitary rectal ulcer are poorly understood. AIMS: To examine the full thickness rectal wall in these two conditions. METHODS: The pathological abnormalities in the surgically resected rectal wall were studied from nine patients with solitary rectal ulcer syndrome, 11 complete rectal prolapse, and nine cancer controls. Routine haematoxylin and eosin and Van Gieson staining for collagen were performed. RESULTS: The rectal wall from solitary rectal ulcer syndrome specimens was thickened compared with complete rectal prolapse and controls. The major difference was in the muscularis propria (2.2 v 1.1 v 1.2 mm, medians, p < 0.005) and particularly the inner circular muscular layer, and to a lesser extent the submucosal and outer longitudinal muscular layers. Some solitary rectal ulcer syndrome specimens showed unique features such as decussation of the two muscular layers (four of nine), nodular induration of inner circular layer (four of nine) and grouping of outer longitudinal layer into bundles (three of nine); these were not seen in complete rectal prolapse or control specimens. CONCLUSIONS: These features, which resemble the features of high pressure sphincter tissue, may be of aetiological importance, and suggest a different pathogenesis for these two disorders. Excess collagen was seen in both disorders, was more severe in solitary rectal ulcer syndrome specimens, and probably reflects a response to repeated trauma. PMID- 8707094 TI - Faecal phytic acid and its relation to other putative markers of risk for colorectal cancer. AB - AIMS: Phytic acid, a major constituent of cereals, pulses, and seeds has been advocated as an important antioxidant component of dietary fibre that affords possible protection against colorectal cancer. This is supported by experimental studies showing it has antineoplastic activity in animal models of both colon and breast cancer. To date the concentration of faecal phytic acid in human clinical groups has not been evaluated. Therefore the faecal phytic acid content of adenoma patients drawn from a placebo controlled calcium intervention trial was evaluated. METHODS: Phytic acid was measured in faecal extracts by an improved ion-pair high performance liquid chromatography method. RESULTS: Phytic acid was detected in the range 0.68-4.00 mumol/g wet faeces and 55-2038 mumol/day. Linear regression analyses showed no association between stool phytic acid and lipid content. Strong correlations were seen, however, between phytic acid and iron content, both on a concentration (r = 0.52; p = 0.00004) and daily excretion (r = 0.76; p = 5.5 x 10(-12) basis. Phytic acid was also strongly correlated with the daily excretion of calcium (r = 0.59; p = 1.36 x 10(-6) and magnesium (r = 0.42; p = 0.001). Cell proliferation in the sigmoid colon, an intermediate biomarker of colorectal cancer was not significantly associated with faecal phytic acid, minerals or lipid content in this compromised clinical group. CONCLUSIONS: This improved method, developed for the determination of phytic acid in faeces should allow further studies on the role of phytic acid in the aetiology of colorectal cancer to be conducted on a population or case control basis. PMID- 8707095 TI - Decrease in sensitisation rate and intestinal anaphylactic response after nitric oxide synthase inhibition in a food hypersensitivity model. AB - BACKGROUND: Although nitric oxide (NO) has been found to have a role in gut inflammation and to modulate immunoglobulin production, little is known about its part in food hypersensitivities. AIM: This study aimed to evaluate the role of NO through the inhibition of constitutive and inducible NO synthase (cNOS and iNOS respectively) on the sensitisation process (antibody titres) and on intestinal anaphylactic responses (colonic hypersecretion upon antigen challenge). ANIMALS AND METHODS: Guinea pigs sensitised to cow's milk proteins were treated either during the sensitisation period or before antigen challenge by N-nitro-L-arginine methyl ester (L-NAME) (inhibiting both cNOS and iNOS) or amino-guanidine (selective iNOS inhibitor). RESULTS: Chronic treatment by L-NAME or aminoguanidine reduced antibody titres and the secretory response to antigen challenge. In contrast, only L-NAME administered before challenge was able to antagonise the hypersecretion induced by the challenge. CONCLUSIONS: NO generated by iNOS has a role in the sensitisation process: iNOS inhibition results in lower rates of antibodies leading to a reduced secretory response upon challenge. In contrast, blockade of colonic hypersecretion by L-NAME but not by aminoguanidine suggests that NO via cNOS is a key mediator in intestinal anaphylactic reactions. PMID- 8707096 TI - Low dose alpha interferon therapy can be effective in chronic active hepatitis C. Results of a multicentre, randomised trial. AB - BACKGROUND: There is some controversy concerning the efficacy of low dose alpha interferon therapy in chronic hepatitis C. AIMS: To evaluate the effectiveness of treatment with low doses of alpha interferon in chronic hepatitis C. PATIENTS: One hundred and forty one patients with anti-HCV positive chronic active hepatitis C from six hospitals were enrolled in the study. METHODS: Patients were randomised to treatment with 5 MU (group A) or 1.5 MU (group B) injections. The dose was reduced in responders from group A or increased in non-responders from group B to maintain treatment with the minimal effective dose. Patients were treated for 48 weeks and followed up for 24 additional weeks with no treatment. Normalisation of alanine aminotransferase (ALT) was used to evaluate response. RESULTS: A sustained response was seen in eight patients from group A (12%) and in 15 (21%) from group B. This difference was not statistically significant. Increasing the dose of interferon led to sustained response in only five of 58 patients (9%) from group B who did not respond to 1.5 MU injections. In contrast, 15 of 21 patients (71%) in whom ALT remained normal with 1.5 MU injections developed a sustained response. By multivariate analysis sustained response seemed associated with young age and was more frequent in patients with genotype 3 HCV infection. Sustained response was preceded by a rapid normalisation of ALT and was inversely related to the amount of alpha interferon necessary to maintain ALT at low values during treatment. CONCLUSIONS: Some patients with chronic hepatitis C are very sensitive to alpha interferon and can be successfully treated with low doses. Treatment with higher doses may be effective in a minority of patients who do not respond to low doses. PMID- 8707097 TI - Natural history and prognostic factors in 305 Swedish patients with primary sclerosing cholangitis. AB - BACKGROUND/AIMS--The course of primary sclerosing cholangitis (PSC) is highly variable and unpredictable. This study describes the natural history and outcome of PSC. These data were used to construct a prognostic model for patients with PSC. METHODS--A total of 305 Swedish patients with PSC were studied. The median follow up time was 63 (1-194) months and all patients could be traced for follow up. Some 79 patients died or had a liver transplant. The prognostic significance of clinical, biochemical, and histological findings at the time of diagnosis were evaluated using multivariate analysis. RESULTS--The estimated median survival from time of diagnosis to death or liver transplantation was 12 years. Cholangiocarcinoma was found in 24 (8%) of the patients and 134 (44%) of the patients were asymptomatic at the time of diagnosis. The estimated survival rate was significantly higher in the asymptomatic group (p < 0.001). However, 29 (22%) of the asymptomatic patients became symptomatic during the study period. It was found that age, serum bilirubin concentration, and histological stage at the time of diagnosis were independent predictors of a bad prognosis. These variables were used to construct a prognostic model. CONCLUSIONS--This prognostic model developed from a large homogeneous population of PSC patients should be of value for the timing of transplantation and patient counselling in PSC. PMID- 8707098 TI - Granulomatous hepatitis caused by Bacillus Calmette-Guerin (BCG) infection after BCG bladder instillation. AB - BACKGROUND: Bladder instillations with Bacillus Calmette-Guerin (BCG) are commonly used as immunotherapy for bladder carcinoma. Sometimes patients experience serious systemic side effects, such as sepsis or pneumonitis. Granulomatous hepatitis is a rare serious side effect, which has been considered a hypersensitivity reaction to BCG. PATIENT: The first case of granulomatous hepatitis after BCG bladder instillation in which mycobacteria were identified by staining techniques and mycobacterial DNA was detected in liver tissue using the polymerase chain reaction is reported. CONCLUSION: The granulomatous hepatitis was caused by BCG infection of the liver after haematogenous dissemination of BCG, rather than hypersensitivity. PMID- 8707099 TI - Colitis complicating chronic granulomatous disease. A clinicopathological case report. AB - BACKGROUND: This report concerns a female patient now aged 24 years, diagnosed at the age of 7 years as suffering from chronic granulomatous disease. At age 20 she developed diarrhoea accompanied by rectal bleeding. Endoscopy showed extensive colitis. She failed to respond to medical treatment and underwent total colectomy two years later. AIMS: To discuss the histological changes in the colon in chronic granulomatous disease. RESULTS: There was extensive mucosal inflammation throughout colon and rectum resembling ulcerative colitis. In addition characteristic large pigmented macrophages were distributed in the basal mucosa and superficial submucosa. Similar cells accompanied by granulomata were present in mesenteric lymph nodes. CONCLUSIONS: Colitis is an unusual clinical manifestation of chronic granulomatous disease but the presence and characteristic distribution of such pigmented macrophages in colonic biopsy in children and young adults may suggest the diagnosis. PMID- 8707101 TI - Lymphocytic gastritis and coeliac disease. PMID- 8707100 TI - Delta 4-3-oxosteroid 5 beta-reductase deficiency: failure of ursodeoxycholic acid treatment and response to chenodeoxycholic acid plus cholic acid. AB - BACKGROUND: In some infants with liver disease, 3-oxo-delta 4 bile acids are the major bile acids in urine, a phenomenon attributed to reduced activity of the delta 4-3-oxosteroid 5 beta-reductase required for synthesis of chenodeoxycholic acid and cholic acid. These patients form a heterogeneous group. Many have a known cause of hepatic dysfunction and plasma concentrations of chenodeoxycholic acid and cholic acid that are actually greater than those of the 3-oxo-delta 4 bile acids. It is unlikely that these patients have a primary genetic deficiency of the 5 beta-reductase enzyme. AIMS: To document the bile acid profile, clinical phenotype, and response to treatment of an infant with cholestasis, increased plasma concentrations of 3-oxo-delta 4 bile acids, low plasma concentrations of chenodeoxycholic acid and cholic acid, and no other identifiable cause of liver disease. PATIENTS: This infant was compared with normal infants and infants with cholestasis of known cause. METHODS: Analysis of bile acids by liquid secondary ionisation mass spectrometry and gas chromatography-mass spectrometry. RESULTS: The plasma bile acid profile of the patient was unique. She had chronic cholestatic liver disease associated with malabsorption of vitamins D and E and a normal gamma-glutamyltranspeptidase when the transaminases were increased. The liver disease failed to improve with ursodeoxycholic acid but responded to a combination of chenodeoxycholic acid and cholic acid. CONCLUSION: Treatment of primary 5 beta-reductase deficiency requires the use of bile acids that inhibit cholesterol 7 alpha-hydroxylase. PMID- 8707102 TI - Concentrations of epidermal growth factor in human saliva and gastric juice. PMID- 8707103 TI - Helicobacter pylori infection density and gastric inflammation in duodenal ulcer and non-ulcer subjects. PMID- 8707104 TI - Gut moves towards the 3rd millennium. PMID- 8707105 TI - Why do cholestatic patients itch? PMID- 8707106 TI - Non-steroidal anti-inflammatory drugs and the chemoprevention of colorectal and oesophageal cancers. PMID- 8707107 TI - Relation between oesophageal acid exposure and healing of oesophagitis with omeprazole in patients with severe reflux oesophagitis. AB - BACKGROUND/AIMS: Reducing oesophageal acid exposure by suppressing acid secretion with omeprazole is highly effective in healing reflux oesophagitis. Some patients with severe oesophagitis, fail to heal and whether this results from inadequate acid suppression or other factors is unclear. The aim of this study, was to investigate the relation between oesophageal acid exposure and healing in patients with severe reflux oesophagitis treated with omeprazole. METHODS: Sixty one patients with grade 3 or 4 ulcerative oesophagitis were treated for eight weeks with omeprazole 20 mg every morning. Those patients unhealed at eight weeks were treated with 40 mg every morning for a further eight weeks. Endoscopy and 24 hour oesophageal pH monitoring were performed before treatment and at the end of each treatment phase while receiving treatment. RESULTS: Thirty per cent of patients failed to heal with the 20 mg dose. Unhealed patients had greater total 24 hour oesophageal acid exposure before treatment, and while receiving treatment also had greater acid exposure and a smaller reduction in acid exposure than did patients who healed. Forty seven per cent of the unhealed patients also failed to heal with the 40 mg dose. These patients had similar levels of acid exposure before treatment to those who healed, but had greater acid exposure while receiving treatment, particularly at night when supine. CONCLUSIONS: Patients with severe ulcerative oesophagitis who are refractory to omeprazole have greater oesophageal acid exposure while receiving treatment than responding patients. This is due to a reduced responsiveness to acid suppression, and is likely to be an important factor underlying the failure of the oesophagitis to heal. PMID- 8707108 TI - High amplitude contractions in the middle third of the oesophagus: a manometric marker of chronic alcoholism? AB - BACKGROUND: Oesophageal motor abnormalities have been reported in alcoholism. AIM: To investigate the effects of chronic alcoholism and its withdrawal on oesophageal disease. PATIENTS: 23 chronic alcoholic patients (20 men and three women; mean age 43, range 23 to 54). METHODS: Endoscopy, manometry, and 24 hour pH monitoring 7-10 days and six months after ethanol withdrawal. Tests for autonomic and peripheral neuropathy were also performed. Motility and pH tracings were compared with those of age and sex matched control groups: healthy volunteers, nutcracker oesophagus, and gastro-oesophageal reflux disease. RESULTS: 14 (61%) alcoholic patients had reflux symptoms, and endoscopy with biopsy showed oesophageal inflammation in 10 patients. One patient had an asymptomatic squamous cell carcinoma. Oesophageal motility studies in the alcoholic patients showed that peristaltic amplitude in the middle third was > 150 mm Hg (95th percentile (P95) of healthy controls) in 13 (57%), the ratio lower/ middle amplitude was < 0.9 in 15 (65%) (> 0.9 in all control groups), and the lower oesophageal sphincter was hypertensive (> 23.4 mm Hg, P95 of healthy controls) in 13 (57%). All three abnormalities were present in five (22%). Abnormal reflux (per cent reflux time > 2.9, P95 of healthy controls) was shown in 12 (52%) alcoholic patients, and was unrelated to peristaltic dysfunction. Subclinical neuropathy in 10 patients did not effect oesophageal abnormalities. Oesophageal motility abnormalities persisted at six months in six patients with ongoing alcoholism, whereas they reverted towards normal in 13 who remained abstinent; reflux, however, was unaffected. CONCLUSIONS: Oesophageal peristaltic dysfunction and reflux are frequent in alcoholism. High amplitude contractions in the middle third of the oesophagus seem to be a marker of excessive alcohol consumption, and tend to improve with abstinence. PMID- 8707109 TI - Eradication of Helicobacter pylori in patients with duodenal ulcer lowers basal and peak acid outputs to gastrin releasing peptide and pentagastrin. AB - BACKGROUND: Patients with duodenal ulcer (DU) have high basal (BAO) and peak (PAO) acid outputs. The effect of Helicobacter pylori eradication on these variables is unclear. AIM: To discover if gastric acid hypersecretion in patients with DU is caused by H pylori. PATIENTS AND METHODS: BAO, gastrin releasing peptide (GRP), and pentagastrin stimulated PAO in 10 H pylori negative controls, and in 10 H pylori positive patients with DU was measured before and six months after H pylori eradication. H pylori status was determined by histology, culture, and by the 13C-urea breath test. After collecting a 30 minute basal aspirate, GRP 40 pmol/kg/h was infused for 45 minutes, and after a 30 minute washout, pentagastrin 6 micrograms/kg was injected intramuscularly. RESULTS: Basal and stimulated acid output (PAOGRP and PAOPg) were significantly higher in H pylori positive DU than in H pylori negative controls. Six months after H pylori eradication, basal and stimulated acid outputs were all significantly lower than before H pylori eradication. CONCLUSIONS: This study has shown that BAO, PAOGRP, and PAOPg are higher in H pylori positive DU than in H pylori negative controls. All decreased significantly six months after H pylori eradication, to fall within the range of controls. These results are compatible with a hypothesis that acid hypersecretion in duodenal ulcer disease is caused by H pylori infection. PMID- 8707110 TI - Oxyntic endocrine cells of hypergastrinaemic patients. Differential response to antrectomy or octreotide. AB - BACKGROUND/AIMS: To evaluate the response of endocrine cells of the gastric oxyntic mucosa in hypergastrinaemic patients to either antrectomy or treatment with the somatostatin analogue octreotide. PATIENTS: (a) Two patients with enterochromaffin-like (ECL) cell carcinoid and chronic atrophic gastritis, treated with antrectomy; (b) four patients with Zollinger-Ellison syndrome, treated with octreotide. METHODS: Oxyntic endocrine cells were examined by ultrastructural morphometry on full thickness biopsy specimens taken: (a) before and four months after antrectomy, (b) before and after three months' treatment with octreotide 200 micrograms daily. RESULTS: Both treatments induced prompt, significant reduction of gastrinaemia and a significant decrease of the volume density of the whole endocrine cell mass and of the cross sectional area of all nucleated endocrine cell profiles (antrectomy: -38%, p < 0.04 and -31%, p < 0.04, respectively; octreotide: -59%, < 0.007 and -26%, < 0.04, respectively). Assessment of the relative proportion of individual endocrine cell types showed a different response to antrectomy or octreotide. After antrectomy, in fact, only the volume fraction of ECL cells was significantly reduced, from 56.5% to 22.5% ( 60%, p < 0.04). After octreotide treatment, in contrast, the proportion of all endocrine cell types remained remarkably constant, showing that all cell types took part in the observed overall decrease. CONCLUSIONS: Postantrectomy reduction of oxyntic endocrine cells mostly reflects the withdrawal of the specific trophic stimulus of hypergastrinaemia on ECL cells. In contrast, the inhibitory response to octreotide seems to be exerted on virtually all types of oxyntic endocrine cells, probably reflecting a universal occurrence of somatostatin receptors. PMID- 8707111 TI - Prevalence of metronidazole resistant Helicobacter pylori strains among Chinese peptic ulcer disease patients and normal controls in Hong Kong. AB - BACKGROUND: A study was conducted to evaluate the prevalence of metronidazole resistant Helicobacter pylori strains among the Chinese in Hong Kong. The efficacy of the triple therapy that contains metronidazole as one of the anti microbial agents in eradication of the metronidazole susceptible and the metronidazole resistant strains was also assessed. METHODS: Culture for H pylori was attempted from antral biopsy specimens of 70 peptic ulcer and 51 control subjects. Successfully cultured H pylori strains were tested for metronidazole susceptibility. Twenty six peptic ulcer disease subjects who had received a course of triple therapy were also reassessed four to six weeks later for successful eradication of H pylori infection. RESULTS AND CONCLUSIONS: H pylori was successfully cultured from antral biopsy specimens in 69 of 80 (86%) of the infected subjects. The overall metronidazole resistance rate was 53.5% (37 of 69). There was a significantly higher metronidazole resistance rate among H pylori isolates from the asymptomatic controls (20 of 25) than the peptic ulcer disease subjects (17 of 44) (p = 0.0007). Twenty three of 32 (73%) women and 14 of 37 (38%) men harboured the metronidazole resistant strains. There was no sex or age difference as far as the prevalence of metronidazole resistant strains were concerned within each study group. Pre-treatment metronidazole susceptible H pylori were significantly more likely to respond to the triple therapy used than those with the metronidazole resistant ones (14 of 15 v five of 10) (p = 0.021). PMID- 8707112 TI - Influence of cell interactions in a novel model of postnatal mucosal regeneration. AB - BACKGROUND AND AIMS: Conventional models of postnatal mucosal regeneration are cumbersome and limited: a novel model is described here. In addition, the influence of cell interactions on mucosal regeneration is examined within the model. METHODS: Postnatal rat small intestinal mucosa was digested by enzymes to yield heterotypic cell aggregates (CA). CA colony forming ability, growth, and limited cytodifferentiation were assessed in vitro. CA were transplanted subcutaneously and retrieved for histological examination at staggered intervals to assess neomucosal morphogenesis and cytodifferentiation in vivo. Cell interactions in CA were disrupted by enzymes, thus producing cell suspensions (CS). Regeneration by CA and CS were compared. RESULTS: CA produced proliferative colonies in vitro and showed a temporal sequence of neomucosal morphogenesis and differentiation in vivo. CA colonies were more numerous within 24 hours of primary culture and had greater cellularity by 96 hours than CS colonies. Alkaline phosphatase was expressed only by 258 of 696 CA colonies (37%). CA subcutaneous grafts (48 of 56 (87%)) regenerated small intestinal neomucosa while CS were unsuccessful. CONCLUSION: These methods provide a model of mucosal regeneration which includes constituent processes of colony formation, growth, neomucosal morphogenesis, and cytodifferentiation. Preservation of cell interactions within CA seems advantageous to regeneration within the model. PMID- 8707113 TI - Acute epithelial injury in the rat small intestine in vivo is associated with expanded expression of transforming growth factor alpha and beta. AB - BACKGROUND: Previous studies have shown the importance of transforming growth factors alpha and beta (TGF alpha and TGF beta) in modulating epithelial cell restitution after injury in vitro. AIM: To investigate the role of the growth factors TGF alpha and TGF beta after acute epithelial injury in vivo. METHODS: An in vivo model of phytohaemagglutinin (PHA) induced acute epithelial injury in rat small intestine was used. Epithelial cell turnover was assessed by autoradiography and liquid scintillation counting of thymidine uptake. Expression of TGF alpha and TGF beta was assessed by immunohistochemistry. RESULTS: An expansion of the proliferative compartment and increased turnover of intestinal epithelial cells was seen in rats with PHA induced intestinal epithelial injury. Expression of TGF alpha and TGF beta peptides was shown in both the epithelial cell and lamina propria compartment. Different patterns of TGF alpha and TGF beta expression were seen, however, within the epithelium of rats with acute intestinal injury compared with untreated controls, while the expression of these peptides within the lamina propria was not changed. CONCLUSIONS: These findings suggest that acute intestinal epithelial cell injury in vivo is associated with compensatory changes in expression of TGF alpha and TGF beta in the epithelial cell compartment, while the lamina propria does not seem to be significantly affected. PMID- 8707115 TI - Collagen alteration in an animal model of colonic diverticulosis. AB - BACKGROUND: Changes in the structure and integrity of the colon dependent on collagen content and crosslinkage occur with age. AIMS: This study using an animal model examines colonic collagen content and crosslinkage over the lifetime of rats on fibre deficient and higher fibre diets. METHODS: Two groups of 20 rats were fed either a fibre deficient diet (1.7 g NSP (non-starch polysaccharide)/100 g) or a higher fibre diet (13.3 g NSP/100 g) for 18 months. Diverticula were identified by postmortem examination. Caecal and colonic contents were weighed and assayed for short chain fatty acids. Collagen solubility in weak acid was measured to give an indication of the nature and amount of crosslinks in the collagen of the bowel wall. RESULTS: The incidence of colonic diverticula was greater (42.1% fibre deficient rats; 0% higher fibre rats). Colonic collagen solubility index in fibre deficient rats was significantly lower than higher fibre diet fed rats (p < 0.001 in all four sections of the large bowel). Rats with diverticula had the lowest solubility index (p < 0.001 in all four sections of the large bowel). Higher fibre diet rats had increased caecal and colonic contents, caecal and colonic tissue wet weights, and greater caecal short chain fatty acids. Fibre deficient diet fed rats had more pathological abnormalities. CONCLUSIONS: This animal model permits a study of the relation between collagen crosslinkage and the development of colonic diverticulosis. A higher fibre diet protects against collagen crosslinking and this is related to a decreased incidence of diverticula. PMID- 8707114 TI - Small intestinal motor abnormalities in patients with functional dyspepsia demonstrated by ambulatory manometry. AB - AIMS/METHODS: In 30 patients with functional dyspepsia and in 20 healthy volunteers, ambulatory duodenojejunal manometry was performed to examine the interdigestive and postprandial small intestinal motility patterns in relation to symptoms. RESULTS: In the fasting state, the number of migrating motor complex cycles mean (SEM) was significantly lower in patients, especially in patients with dysmotility-like dyspepsia, than in control subjects (3.8 (0.4), 2.6 (0.5), and 5.3 (0.7) cycles, respectively; p < 0.05), due to a longer duration of phase II. Non-propagated and retrogradely propagated phase III activity was more prevalent in patients than in control subjects (48% v 15%; p = 0.020). During phase II and after dinner no differences were found in contraction incidence, mean amplitude or motility index. However, 1 1/2 hours after completing breakfast the motility index was higher in patients at all three recording levels (p < 0.05). Burst activity was more prevalent in patients than in control subjects (22% v 6% of the subjects; p = 0.003). In 41% of the patients the symptom index was > 75%. CONCLUSIONS: These results suggest that small intestinal motor abnormalities, especially during fasting, participate in the pathogenesis of symptoms in patients with functional dyspepsia. Ambulatory manometry of the small intestine is a valuable tool to demonstrate these abnormalities in outpatients pursuing their daily activities. PMID- 8707116 TI - Sulindac increases the expression of APC mRNA in malignant colonic epithelial cells: an in vitro study. AB - BACKGROUND: Sulindac is a non-steroidal anti-inflammatory drug which induces regression of colonic polyps in patients with familial adenomatous polyposis. Animal and in vitro studies have shown that both the sulphide metabolite of sulindac, which is able to inhibit cyclo-oxygenase, and the sulphone metabolite, which lacks this ability, are able to inhibit the growth of colonic carcinoma cells. The exact mechanism by which these effects occurs is not known. AIMS: To examine the effect of sulindac sulphide and sulindac sulphone on the expression of APC messenger RNA (mRNA), and on the proliferation of colonic carcinoma cells in vitro. METHODS: The colonic carcinoma cell line LIM 1215 was treated with sulindac sulphide and sulindac sulphone (10 microM or 100 microM) for 24 hours. Total RNA was extracted and APC mRNA was quantitated using competitive reverse transcription polymerase chain reaction. Measurements of cell number, cell proliferation, and prostaglandin E2 concentrations were also made. RESULTS: A significant increase in APC mRNA was observed after treatment with 10 microM of both sulindac sulphide and sulindac sulphone (control: 37.2 (19.7); 10 microM sulindac sulphide: 129 (112.8); 10 microM sulindac sulphone: 207.7 (102.9) pg/(g total RNA) (p < 0.05). Prostaglandin E2 concentrations were significantly reduced after treatment with sulindac sulphide, but not after sulindac sulphone. Both agents produced a dose dependent reduction in cell numbers and cell proliferation, which was more noticeable after treatment with sulindac sulphide. CONCLUSIONS: Both sulindac sulphide and sulindac sulphone inhibit the growth of carcinoma cells in vitro and cause an increase in APC mRNA. The effect of these agents on colonic carcinogenesis is not mediated entirely by means of an inhibition of prostaglandin biosynthesis. PMID- 8707117 TI - Smoking, hypertension, and colonic anastomotic healing; a combined clinical and histopathological study. AB - BACKGROUND: Large bowel anastomotic breakdown occurs as a result of perianastomotic ischaemia. Preservation of the macroscopic arterial supply to the perianastomotic tissues is vital, but little is known about the influence of microvascular disease on anastomotic healing. AIMS: To study the associations between risk factors for macrovascular disease, the presence of colonic microvascular disease, and the incidence of anastomotic dehiscence. PATIENTS: 147 consecutive colonic surgery patients. METHODS: The prevalence of smoking, hypertension, diabetes, and ischaemic heart disease were established retrospectively from patient notes. These risk factors were correlated with histopathological assessment of resection margin vasculature and clinical follow up. RESULTS: Smoking and hypertension were significantly associated with an increased incidence of anastomotic dehiscence and microvascular disease. Microvascular disease was positively correlated with an increased incidence of anastomotic dehiscence. CONCLUSIONS: Microvascular disease predisposes to anastomotic breakdown. This effect may in part be due to vasospasm in the diseased vessels, which are hypersensitive to serotonin, a vasoactive amine known to be present in increased quantities in the serum of smokers, hypertensives, and after surgery. Treatment with serotonin antagonists in the perioperative period may be beneficial to anastomotic healing, helping to maintain microvascular flow. PMID- 8707118 TI - Bile acid induced colonic irritation stimulates intracolonic nitric oxide release in humans. AB - AIM: To measure the intracolonic release of nitric oxide end products (nitrates plus nitrites) and eicosanoids in response to intraluminal irritation with deoxycholic acid (DCA). PATIENTS: Seven patients with irritable bowel syndrome. METHODS: The left colon was perfused with a solution with or without 3 mM deoxycholic acid. Aspirates were assayed for eicosanoids by specific radioimmuno assay, and for nitrates plus nitrites by the Griess reaction. To confirm that stimulated colonic mucosa can produce nitric oxide (NO), ancillary studies were performed in vitro using samples of normal mucosa obtained from five surgically resected colons. Samples were incubated for 30 minutes in Kreb's solution, 3 mM DCA or DCA with 1 mM L-nitro-arginine-methyl-ester (L-NAME) to inhibit the NO synthase. Finally, NO synthase activity was measured in five samples of human colonic mucosa. RESULTS: Intracolonic release of nitrates plus nitrites was basally undetectable in six of seven patients. Bile acid considerably increased the release of prostaglandin E2 and nitrates plus nitrites (p < 0.01). By contrast, no increase in thromboxane and leukotriene was seen. In vitro mucosal incubation with DCA increased the production of NO synthase products, which was blocked by L-NAME. Activity of Ca+2 independent NO synthase was detectable in four of five samples of human colonic mucosa. CONCLUSION: The human colonic mucosa responds to bile acid induced irritation by a surge in NO generation via NO synthase. PMID- 8707119 TI - Pathogenesis of aphthoid ulcers in Crohn's disease: correlative findings by magnifying colonoscopy, electron microscopy, and immunohistochemistry. AB - BACKGROUND: The mechanism of ulceration in Crohn's disease remains unknown. AIMS: To clarify the role of the follicle associated epithelium (FAE) of colonic lymphoid nodules in the formation of ulcers in Crohn's disease. METHODS: After identification of colonic lymphoid nodules and aphthoid lesions by magnifying colonoscopy, 76 biopsy specimens were obtained from 10 patients with Crohn's disease and three patients with colonic lymphoid hyperplasia. This study correlated magnifying colonoscopic, electron microscopic, and immunohistochemical findings of biopsy specimens. RESULTS: In Crohn's disease, scanning electron microscopy of lymphoid nodules surrounded by a red halo without visible erosions by magnifying colonoscopy, showed surface erosions 150-200 microns in size. These lymphoid nodules with red halos had small erosions either light microscopically or electron microscopically in 18 of 21 specimens (86%). Correlation of scanning and transmission electron microscopy showed residues of FAE including M cells at the edges of the erosions. In immunohistochemical studies, HLA-DR antigen was limited in M cells of FAE in the patients with lymphoid hyperplasia without inflammatory bowel disease. In Crohn's disease patients in remission, however, HLA-DR antigen was strongly expressed over the entire FAE of lymphoid nodules with a red halo endoscopically, while the expression was weak and irregular in the mucosa surrounding the lymphoid nodules. HLA-DR was strongly expressed in the entire inflamed colonic mucosa in the active stage. CONCLUSION: The red halo appearance surrounding lymphoid follicles seems to precede visible aphthoid ulcers and suggests that ulcerations in Crohn's disease originate from FAE, possibly related to its physiological role as a portal of entry for potentially pathogenic agents. PMID- 8707120 TI - Thrombotic vascular risk factors in inflammatory bowel disease. AB - BACKGROUND: Thrombosis may be an important effector mechanism in the pathogenesis of Crohn's disease. METHODS: This study therefore investigated the prevalence of independent thrombotic risk factors (factor VII coagulant activity, lipoprotein (a), fibrinogen, plasma triglycerides, and smoking) in patients with Crohn's disease, ulcerative colitis, and normal controls. RESULTS: In Crohn's disease (n = 75), the mean plasma VII:C, lipoprotein (a) and fibrinogen concentrations were significantly greater than in the normal population (n = 85). In ulcerative colitis (n = 35), only the mean factor VII:C concentration was significantly higher than normal. Ninety three per cent of patients with Crohn's disease and 86% of those with ulcerative colitis had at least one risk factor for thrombotic vascular disease, compared with 61% of the normal population (p < 0.001). CONCLUSIONS: In many young patients with inflammatory bowel disease, plasma concentrations of these prothrombotic factors were in excess of the limits that are regarded as posing an increased risk for the development of occlusive vascular disease. PMID- 8707121 TI - Clinical patterns of familial inflammatory bowel disease. AB - BACKGROUND: Although many recent studies have shown the increased risk of inflammatory bowel disease in relatives of patients with Crohn's disease and ulcerative colitis, clinical patterns of disease within families remain relatively poorly documented. AIMS: In this study, clinical characteristics (disease type, extent, age on onset, need for surgery, and presence of extraintestinal manifestations) have been compared in affected subjects in multiply-affected families, with inflammatory bowel disease. METHODS: 54 families in whom one parent and at least one child were affected (a total of 77 parent child pairs) and 155 families in whom at least two siblings were affected (a total of 190 affected sibling pairs) were involved. RESULTS: In affected parent child pairs, parent and child were concordant for disease type in 58 of 77 pairs (75.3%), for extent in 63.6%, extraintestinal manifestations in 70.1%, and smoking history in 85%. The median age of onset in parents was significantly higher than offspring (p < 0.0001). In 40 pairs, 60.6%, the parent was at least 10 years older than child. Siblings were concordant for disease type in 81.6% of the affected sibling pairs, extent in 76.0%, extraintestinal manifestations in 83.8%, and smoking history in 81.3%. In contrast with the parent-child pairs, 68.1% (111 sibling pairs) siblings were diagnosed within 10 years of each other. The median age of onset was 24.0 years. CONCLUSIONS: This study has shown consistent clinical patterns in many families with inflammatory bowel disease. The differences in age of onset between parents and children are not readily explained by a simple cohort effect or ascertainment bias, and may reflect effects of genetic factors, producing anticipation between generations. PMID- 8707122 TI - Acute ethanol administration induces oxidative changes in rat pancreatic tissue. AB - BACKGROUND: There is mounting clinical evidence that ethanol toxicity to the pancreas is linked with glutathione depletion from oxidative stress but there is not experimental proof that this occurs. AIMS AND METHODS: The effect of acute ethanol ingestion (4 g/kg) on the pancreatic content of reduced (GSH) and oxidised (GSSG) glutathione, malondialdehyde (MDA), and carbonyl proteins were therefore studied in the rat. RESULTS: Ethanol caused a significant reduction in GSH (p < 0.02) and an increase in GSSG (p < 0.005), MDA (p < 0.05), and carbonyl proteins (p < 0.05) in the rat pancreas. The GSH/GSSG ratios were significantly decreased after ethanol, especially in rats pretreated with diethylmaleate (DEM), a GSH blocker. Administration of ethanol after DEM further increased the rate of lipid and protein oxidation. Pretreatment with cyanamide (an inhibitor of aldehyde dehydrogenase) but not with 4-methylpyrazole (an alcohol dehydrogenase inhibitor) caused higher production of GSSG and MDA. CONCLUSIONS: These findings indicate that acute ethanol reduces the pancreatic content of GSH, which seems to be protective against ethanol toxicity, since its depletion is accompanied by increased oxidative damage to cell structures. The further increase of lipid peroxidation and GSSG production in the presence of cyanamide suggests that acetaldehyde might be responsible for the oxidative changes that occur in pancreatic cells after ethanol administration. PMID- 8707123 TI - Lipopolysaccharide induced apoptosis of rat pancreatic acinar cells. AB - BACKGROUND: Bacterial lipopolysaccharide (LPS) has been proposed to participate in the pathogenesis of pancreatic inflammatory disease. AIMS: This study investigated the role of endotoxaemia in the pathogenesis of pancreatic acinar cell injury. METHODS: Sixty eight male Spraque-Dawley rats were used in the study. Escherichia coli LPS (5 mg/kg) was injected into the peritoneal cavity of the rats. The concentration of pancreatic phospholipase A2 (PLA2) in plasma was measured and pancreatic tissue examined by histology, in situ detection of free DNA 3'-ends, and electrophoretic DNA analysis. RESULTS: The concentration of pancreatic PLA2 increased in plasma and the catalytic activity of PLA2 increased in pancreatic tissue after an LPS injection. Apoptosis in pancreatic acinar cells and fragmentation of DNA typical of apoptosis in pancreatic tissue was seen 24 hours after an LPS injection. Pancreatic acinar atrophy was seen 72 hours after the LPS injection. CONCLUSIONS: These data show that LPS causes release of pancreatic PLA2 into blood plasma, activation of PLA2 in pancreatic tissue, and apoptosis of acinar cells. PMID- 8707125 TI - Role of paraoesophageal collaterals and perforating veins on outcome of endoscopic sclerotherapy for oesophageal varices: an endosonographic study. AB - BACKGROUND: Endoscopic sclerotherapy (EST) is an established method for controlling and preventing bleeding from oesophageal varices. However, oesophageal varices sclerose easily and require less sessions of EST in some patients while few fail to respond. This study therefore looked at changes in the vascular anatomy of the lower oesophagus and upper stomach that accompany successful sclerotherapy of oesophageal varices. METHODS: Endoscopic ultrasonography was performed in 50 patients with cirrhotic portal hypertension before (control, 20 patients) and after successful obliteration of varices with endoscopic sclerotherapy in a group of responders (EST-R, 20 patients) and in a group of non-responders (EST-NR, 10 patients). RESULTS: The median number and size of submucosal veins at the gastrooesophageal junction and in the lower oesophagus were significantly less in the EST-R group compared with control and EST-NR groups (p values between < 0.00001 and < 0.000001). Concomitantly, the number and size of paraoesophageal collaterals were significantly less in the EST R group compared with the other two groups (p values between 0.02 and 0.00007). Perforating veins were identified in 14 (70%) patients in the control group, nine (90%) in the EST-NR group and in none in the EST-R group (p < 0.001 for both controls v EST-R and EST-R v EST-NR, and p = NS, control v EST-NR). CONCLUSION: Oesophageal variceal sclerosis is associated with significant reduction in the number and size of paraoesophageal collaterals and disappearance of perforating veins in the lower oesophagus. PMID- 8707124 TI - Hyperlipaemia intensifies the course of acute oedematous and acute necrotising pancreatitis in the rat. AB - BACKGROUND: Serum triglyceride concentrations higher than 10 to 20 mmol/l are probably a risk factor for developing acute pancreatitis in humans. AIMS: To therefore analyse the influence of hyperlipaemia on the course of acute oedematous and acute necrotising pancreatitis in rats. SUBJECTS: Male Wistar rats were used in all experiments. METHODS: Six different groups of animals were used: two groups without pancreatitis (controls), two with acute oedematous pancreatitis, and two with acute necrotising pancreatitis. One group from each pair was treated with Triton WR 1339, which induces endogenous hyperlipaemia. Blood samples were taken from all subjects to measure triglyceride, cholesterol, amylase, and lipase. Pancreatic tissue samples were taken and the degree of pancreatic damage was judged microscopically. RESULTS: In the control groups no significant changes occurred, either in serum enzyme activities or in histology. The hyperlipaemic subgroup of animals with acute oedematous pancreatitis developed significantly higher (p < 0.001) serum amylase activities and a greater degree of histological damage (p < 0.01) than the animals of the non hyperlipaemic acute oedematous pancreatitis group. In the animals with necrotising pancreatitis, serum lipase activity and the histological degree of pancreatic damage were significantly higher in the hyperlipaemic animals than in the non-hyperlipaemic animals. CONCLUSION: This study shows that hyperlipaemia intensifies the course of acute oedematous and acute necrotising pancreatitis in rats. PMID- 8707126 TI - A duodenal mucosal abnormality in the reduction of Fe(III) in patients with genetic haemochromatosis. AB - BACKGROUND: Previous in vitro studies have shown that the uptake of Fe(III) by freshly isolated duodenal mucosal biopsy specimens is increased in patients with genetic haemochromatosis. Moreover, in the mouse it has recently been found that reduction of Fe(III) to Fe(II) is a prerequisite for iron uptake by the proximal intestine. AIMS/METHODS: This study used the in vitro technique to investigate the rates of reduction and uptake of 59Fe(III) by duodenal mucosal biopsy specimens obtained at endoscopy from treated and untreated patients with genetic haemochromatosis. RESULTS: The rate of reduction of iron in the medium was proportional to the incubation time and was not caused by the release of reducing factors from the tissue fragments. Ferrozine, a specific Fe(II) chelator and ferricyanide, a non-permeable oxidising agent, inhibited uptake of 59Fe showing that reduction of Fe(III) precedes uptake. The rates (all values given as pmol/mg/min) of reduction (152 (49) v 92 (23)) and uptake (8.3 (4.0) v 3.6 (1.3), mean (SD)), were significantly increased in biopsy specimens from the untreated group (n = 6) compared with those from 10 control subjects (p < 0.04). Furthermore, the reduction and uptake rates were still increased in five patients in whom iron stores were normal after venesection treatment. CONCLUSIONS: These results show that there is a persistent abnormality in the reduction and uptake of iron by the intestine in genetic haemochromatosis. PMID- 8707127 TI - Sclerotherapy versus sclerotherapy and propranolol in the prevention of rebleeding from oesophageal varices: a randomised study. AB - BACKGROUND: This trial was carried out to assess the value of propranolol in the prevention of recurrent variceal bleeding when combined with longterm endoscopic sclerotherapy. PATIENTS AND METHODS: Two hundred patients (161 male, 39 female, age range 20-68 years) with portal hypertension resulting mainly from schistosomal periportal fibrosis or posthepatitic cirrhosis presenting with their first episode of haematemesis or melena, or both were included. This was confirmed endoscopically to result from ruptured oesophageal varices. After initial control of bleeding, patients were randomised into two groups: group 1 treated with endoscopic sclerotherapy alone and group 2 treated with sclerotherapy plus propranolol. They were followed up for two years. RESULTS: Group (2) had a lower rebleeding rate (14.3% v 38.6% in group 1), lower variceal recurrence after obliteration (17% v 34% in group 1), longer period between variceal obliteration and recurrence (36 weeks v 21 weeks in group 1); but no change in mortality (12% in both groups). CONCLUSIONS: Patients treated with sclerotherapy should be given propranolol for longterm management. PMID- 8707128 TI - Roles of gall bladder emptying and intestinal transit in the pathogenesis of octreotide induced gall bladder stones. AB - BACKGROUND: Octreotide treatment of acromegalic patients increases the % deoxycholic acid conjugates and the cholesterol saturation of gall bladder bile, and induces gall stone formation. AIMS: To study the roles of gall bladder emptying and intestinal transit in these phenomena. METHODS AND PATIENTS: Gall bladder emptying and mouth to caecum transit was measured in (a) control subjects and acromegalic patients given saline or 50 micrograms of octreotide, and (b) acromegalic patients taking long term octreotide. In the second group, large bowel transit was also measured. RESULTS: A single dose of octreotide inhibited meal stimulated gall bladder emptying, the ejection fraction falling from mean (SEM) 66.0 (2.3)% to 7.0 (5.3)% in controls (p < 0.001); from 72.5 (2.1) to 16.6 (5.1)% in untreated acromegalic patients (p < 0.001), and to 30.4 (9.5)% in acromegalic patients taking long term octreotide (p < 0.001 v untreated acromegalic group). Octreotide prolonged mouth to caecum transit time, from 112 (15) min to 237 (13) min in controls (p < 0.001), from 170 (13) min to 282 (11) min in untreated acromegalic patients (p < 0.001), and to 247 (10) min in acromegalic patients taking long term octreotide (p < 0.001 v untreated acromegalic patients). The mean large bowel transit in octreotide untreated compared with treated acromegalic patients remained unchanged (40 (6) h v 47 (6) h). CONCLUSIONS: Prolongation of intestinal transit and impaired gall bladder emptying may contribute to lithogenic changes in bile composition and gall stone formation in patients receiving long term octreotide. PMID- 8707129 TI - Obstructive jaundice causes reduced expression of polymorphonuclear leucocyte adhesion molecules and a depressed response to bacterial wall products in vitro. AB - BACKGROUND: Obstructive jaundice is associated with an increased incidence of infection and endotoxaemia, which may result from impaired host immunity. Neutrophil adhesion to vascular endothelium is a key part of the inflammatory response. AIMS: To investigate neutrophil adhesion molecule expression and activation in obstructive jaundice. PATIENTS: Nine adult patients with obstructive jaundice and 11 control subjects. METHODS: The expression of the neutrophil adhesion receptors L-selectin, CD11a, CD11b, CD11c, and CD15 was determined using flow cytometry. CD11b expression in response to stimulation with fMLP and endotoxin was measured. RESULTS: The basal expression of L-selectin, CD11a, and CD15 was significantly decreased in jaundiced patients (p < 0.05) and the expression of CD11b in response to stimulation with fMLP and endotoxin was significantly impaired in the jaundiced group. Endotoxin stimulation without plasma did not reverse the impaired response showing that it is not caused by endotoxin inactivation by plasma proteins. CONCLUSIONS: Neutrophils from patients with obstructive jaundice show decreased adhesion receptor expression and an impaired response to stimulation with bacterial products. This cellular dysfunction may be responsible for the high incidence of septic complications in these patients. PMID- 8707130 TI - Interchange between collagenous and lymphocytic colitis in severe disease with autoimmune associations requiring colectomy: a case report. AB - BACKGROUND: Collagenous colitis and lymphocytic colitis present with a similar clinical picture. Whether these conditions are separate entities or whether they represent different pathological stages of the same condition is an unresolved issue. PATIENT: This is a case of collagenous colitis following a fulminant course in which a colectomy was necessary. In the operative specimen the thickened collagen plate, which had been present only two weeks preoperatively had been lost and the pathology was of a lymphocytic colitis. Six months postoperatively this patient developed a CREST syndrome and primary biliary cirrhosis. CONCLUSIONS: This case shows the lability of the collagen plate and the common ground between collagenous and lymphocytic colitis, and presents evidence that these two conditions are different manifestations of the same disease. It also describes for the first time an association between collagenous colitis and CREST syndrome and primary biliary cirrhosis. PMID- 8707131 TI - Association of Helicobacter pylori infection, lymphoid follicles, and lymphocytic gastritis: a risk factor for the development of primary gastric lymphoma? PMID- 8707132 TI - Colonocyte metabolism. PMID- 8707133 TI - Helicobacter pylori eradication and serum pepsinogens. PMID- 8707135 TI - Every nurse needs a nurse: reconceptualizing models and methods of care. PMID- 8707134 TI - Novel germline APC gene mutation in a large familial adenomatous polyposis kindred displaying variable phenotypes. PMID- 8707136 TI - Parish nursing: ministry of healing. PMID- 8707137 TI - Starting a nurse-managed center for older adults: the needs assessment process. AB - Nursing centers can be a vital part of the plan to meet the health needs of the coming "age wave." Nurses, who provide both the care for chronic illness and the means for preventing the onset of chronic disability, are in key positions to influence the evolving health care system to create new systems of community based care. The time for NMCs is, perhaps, now or never. Conducting a thorough needs assessment is the first important step. PMID- 8707138 TI - Long-term ventilator-dependent care in nursing homes. AB - For some persons, being dependent on a ventilator is the only alternative to death. Most residents residing in New York state's approved ventilator-dependent beds have needed the support of a ventilator for more than a year. The quality of their lives since admission to specialized programs has been significantly different than it had been in hospitals. These residents are now actively participating in the life of the facility. Many who had not spoken during the entire time they were hospitalized are now speaking with the aid of special tracheostomy tubes. Many have been assisted out of bed for their first shower or tub bath since hospitalization. Additionally, each resident participates in activities of his or her choice. If able, residents, at their request, are taken out of the facility to attend family functions. Outcomes of care in these facilities have generally been positive: some residents have been weaned from ventilator use; others have been discharged home. The overwhelming positive aspects of these programs are evinced by individuals living in an environment that promotes the best quality of life and quality of care possible when living at home is no longer possible. PMID- 8707139 TI - Learning is a two-way street: reciprocity and rewards. PMID- 8707140 TI - Special socks for special people: falls in special care units. PMID- 8707141 TI - Tuberculosis and the elderly living in long-term care facilities. PMID- 8707142 TI - The cost-effectiveness of home health: a case presentation. PMID- 8707143 TI - Hospice care and its relationship to home care services: a case study. PMID- 8707144 TI - When food doesn't taste good anymore. PMID- 8707145 TI - Caffeine, nicotine, and drugs. PMID- 8707146 TI - Hospice concepts in the care of end-stage dementia. PMID- 8707147 TI - Society and elder care. PMID- 8707148 TI - Campaigning for breast cancer awareness. PMID- 8707149 TI - Florence Nightingale, where are you? PMID- 8707150 TI - Salicylism in the elderly: "a little aspirin never hurt anybody"! PMID- 8707151 TI - Medications and the visually impaired elderly. PMID- 8707152 TI - Assessing problem feeding behaviors in mid-stage Alzheimer's disease. PMID- 8707153 TI - Recognizing and helping older persons with vision impairments. PMID- 8707154 TI - Sleep problems and self-care in very old rural women: nursing implications. AB - Sleep problems are prevalent among very old rural women. In a effort to manage them, they may resort to the indiscriminate use of OTC medications and alcohol. As part of the comprehensive nursing care needed to address these issues, rural nurses should carefully assess their clients sleep patterns and self-care activities and encourage the use of nonpharmacologic interventions to improve sleep. A knowledge of rural values is essential to this process. PMID- 8707155 TI - When lithium does not help: the use of anticonvulsants and calcium channel blockers in the treatment of bipolar disorder in the older person. AB - Although anticonvulsant agents and calcium channel blockers do not have any clear advantages over lithium, they do offer patients who cannot (or will not) take lithium another treatment option. It is not yet clear from the literature who will respond best to which drug or combination of drugs. The nurse should be supportive to the patients and family, in what may be a drawn out process, to find the best treatment. Optimism is justified because a lack of response to one drug is not indicative of nonresponse to other drugs. It is important to actively treat bipolar disorder because each episode of mania increases the risk of progression of the illness, with increasingly severe episodes occurring closer together. Bipolar disorder has high social costs (legal, financial, and relationship problems) that make improvements in treatment important for the patient and society. Anticonvulsant agents and calcium channel blockers may also be useful in treating depression. The number of people whose depressive symptoms respond is far less (25% to 30%) than the number who respond to the anti-manic effects, but this is an option when antidepressants and electroconvulsive therapy are not effective. PMID- 8707156 TI - Research brief: degree of personalization as a cue in the assessment of adjustment to congregate housing by rural elders. PMID- 8707157 TI - A controlled evaluation of a lifts and transfer educational program for nurses. PMID- 8707158 TI - Well elder clinics: changing a student's views on aging. PMID- 8707159 TI - Diet for Diabetes--new exchange lists. PMID- 8707160 TI - Helping older adults become nicotine free. PMID- 8707161 TI - Symposium on low molecular weight heparins (LMW heparins) PMID- 8707162 TI - Low-molecular-weight heparins and new strategies for the treatment of patients with established venous thrombosis. AB - Unfractionated heparin is the commonest treatment for established venous thromboembolism. While this treatment undoubtedly reduces mortality and morbidity there are problems associated with its use. It does not always prevent thrombus propagation or embolisation, the low bioavailability results in a frequent failure to achieve therapeutic heparin levels in vivo and the variable sensitivity of the partial thromboplastin time to the heparin effect may result in inappropriate heparin dosage. The low-molecular-weight heparins have high predictable bioavailability and can be administered as weight-calculated fixed dose regimens for the treatment of established venous thromboembolism. While statistically significant clinical results are awaited, there is increasing evidence for the superior benefit-risk ratios for these agents compared to unfractionated heparin. In routine practice, the frequent failure to achieve a therapeutic intensity of anticoagulation is currently the main reason for adopting low-molecular-weight heparins for first-line treatment of venous thromboembolism. Cost analysis studies based on total health care costs may support the use of these drugs, because savings from the abolishment of laboratory monitoring, improved clinical outcome and shorter inpatient stay may prove treatment with low-molecular weight heparin to be more cost-effective than treatment with unfractionated heparin. PMID- 8707163 TI - Prevention of venous thromboembolic risk in non-surgical patients. AB - The risk of deep-vein thrombosis in hospitalized medical patients varies considerably among diseases and should be systematically assessed as a function of the presence of different risk factors. The need for drug prophylaxis of thromboembolic risk in patients at moderate or high risk of thrombosis is now recommended. In such cases, only standard or low-molecular-weight heparins have been evaluated in controlled trials. The efficacy of heparin prophylaxis has been demonstrated in hospitalized cardiac patients following myocardial infarction, in neurological patients with paralysis of one or both lower limbs, and in intensive care patients or in those with a serious medical condition, particularly if they have a history of deep-vein thrombosis. The choice between the different heparins, their dosages and treatment durations, and the possible indication for prophylaxis in patients at low thrombotic risk should be evaluated in controlled trials. PMID- 8707164 TI - Pharmacological activities of heparin chains: should our past knowledge be revised? AB - Unfractionated heparin is a heterogeneous mixture of sulphated polysaccharides. The anticoagulant activity of unfractionated heparin is mainly accounted for by its fractions with a sequence with binding affinity for antithrombin III. The interaction of heparin with antithrombin III markedly enhances the ability of the latter to inhibit factor Xa and thrombin. Only about one third of the unfractionated heparin molecules contain the high-affinity material and its distribution along the heparin molecules appears to be random-hence the development of low-molecular-weight heparins. Most of the saccharide chains of low-molecular-weight heparins are composed of less than 18 saccharides and have reduced ability to inhibit thrombin relative to their ability to inhibit factor Xa. The potential dissociation of anti-factor IIa activity and the anti-Xa activity provided the pharmacological background for the clinical development of low-molecular-weight heparins. During the last few years, evidence has accumulated on the importance of anti-factor IIa for the expression of the antithrombotic activity of low-molecular-weight heparins. This paper reviews the evidence and revises our knowledge about the pharmacological activity of the heparin chains. PMID- 8707166 TI - The role of low-molecular-weight heparins in the prevention of venous thrombosis in surgery with special reference to enoxaparin. AB - Prophylaxis with low-dose heparin has contributed significantly to the reduction in thromboembolic complications in surgery. Without prophylaxis, the rate of deep vein thrombosis is about 30% in patients undergoing general surgery, rising up to 70% in orthopedic and trauma surgery. According to Collins et al. [N Engl J Med 1988; 318: 1162-1172] and Clagett and Reisch [Ann Surg 1988; 208: 227-240] heparin prevented at least 60% of deep-vein thromboses. Meanwhile, various heparin fragments have become available for clinical use and the question arises whether these low-molecular-weight heparins are equal or even superior to unfractionated heparin in preventing thromboembolic complications. Because of the pharmacological heterogeneity of low-molecular-weight heparins and variations in administered doses, this question can only be partially answered by meta analysis. However, in summary it can be said that despite the difficulties of and concerns about general assessments, single daily injections of low-molecular weight heparin are at least as efficacious as multiple daily doses of unfractionated heparin. In particular, enoxaparin 40 mg once a day and enoxaparin 30 mg twice a day seem to be superior to unfractionated heparin in high-risk patients, and adverse drug reactions occur less frequently when this prophylaxis is used. PMID- 8707165 TI - Low-molecular-weight heparins: an overview of their pharmacodynamics, pharmacokinetics and metabolism in humans. AB - Low-molecular-weight heparins (LMWHs) comprise a group in the class of antithrombotic medications, a class headed by unfractionated heparin (UFH). The LMWHs, with mean molecular weights of 4.0-6.0 kD, differ in their individual manufacturing processes, the distribution of their fragment molecular weights, their in vitro potency (anti-Xa, antithrombin and anticoagulant activities) and, consequently, in their biodynamic patterns, recommended dose regimen, and efficacy/safety ratio. Their drug disposition profiles have been evaluated using two significant markers of their pharmacodynamic activity, namely anti-Xa and anti-IIa activities. Since they are mainly administered subcutaneously, then compared to UFH, they are almost completely absorbed (F > or = 90%) and, in contrast to UFH, those for which data are available in the literature exhibit linear pharmacokinetics with proportionality between anti-Xa (and anti-IIa in some cases) plasma concentration and dose, and stationary distribution volume and clearance processes when the dosage is increased. Their distribution volume is close to the blood volume, they are partially metabolized by desulphatation and depolymerization, but urinary excretion of anti-Xa activity for enoxaparin, dalteparin and nadroparin, all given at doses for prevention of venous thrombosis, is between 5 and 10% of the injected dose. However, these LMWHs differ in the extent of their non-renal clearance, resulting in different apparent elimination half-life values and relative apparent bioavailability. When considering certain at-risk situations, using doses for preventing thromboembolism, the LMWHs do not significantly cross the placenta of pregnant women and their excretion profiles are only slightly altered in severe (endogenous creatinine clearance less than 15 ml/min) renal disease patients when given at doses recommended for prevention of venous thromboembolism. Because of the differences among LMWHs, the clinical profile of a given LMWH cannot be extrapolated to another one or generalized to the whole LMWH family. PMID- 8707167 TI - The venous thrombotic risk in non-surgical patients: epidemiological data and efficacy/safety profile of a low-molecular-weight heparin (enoxaparin). The Prime Study Group. AB - In a multicentre, randomized, double-blind controlled trial comparing the low molecular-weight heparin enoxaparin (40 mg) with a standard unfractionated heparin (Ca-heparin, 3 x 5,000 U) in deep-vein thrombosis prophylaxis in a high risk group of 959 hospitalized medical patients, enoxaparin was at least as efficacious as standard heparin, with fewer adverse events. PMID- 8707168 TI - Are low-molecular-weight heparins useful for the prophylaxis and treatment of arterial thrombi? AB - The pharmacologic specificity of low-molecular-weight heparins (LMWHs) has enabled multiple attractive developments in the prophylaxis and treatment of arterial thrombosis. Their high antithrombotic potency associated with a potentially lower induced bleeding risk, the lack of platelet interaction, the prevention of myointimal hyperplasia, and the lower incidence of heparin-induced thrombocytopenia, are major advantages. New studies in cardiology and vascular surgery demonstrate a high efficacy for LMWHs associated with a low risk. PMID- 8707169 TI - [Screening for scoliosis in western Galilee schools]. AB - Scoliosis screening is still under debate. It is generally agreed that it is not cost-effective, but some still favor it as integral to preventive medicine. Among 2940 school children, aged 9 - 13, from several Moslem-Arab villages in the western Galilee, 55 (1.9%) were found to have some deformity on physical examination and were referred for spinal X-ray. Of these, 24 were X-rayed and in 20 of them there was a scoliosis of more than 7 degrees. The prevalence of in family marriage in those with deformities was 31%, not significantly different from the 35% found in their communities. Using the worst-case analysis, the Moslem-Arab population of western Galilee cannot be regarded as a population at risk for scoliosis. Considering the poor cooperation of the parents with further investigation, we conclude that scoliosis screening should not be performed in this community. PMID- 8707170 TI - [Magnuson-Stack operation for chronic anterior shoulder instability]. AB - Between the years 1975-1992 we operated on 30 cases of recurrent anterior dislocation of the shoulder by the Magnuson-Stack procedure, using the subscapularis tendon. 23 patients (25 shoulders) have since been followed for an average of 5 years. We developed a comprehensive shoulder rating scale to evaluate functional status of involved shoulders. By this rating system the results were excellent in 73.9% of the cases, good in 17.4% and fair in 8.7%. The operation is technically simple, rapid, had no complications and is at least as effective as other operations. PMID- 8707171 TI - [Epidural analgesia for postoperative pain in infants and children]. AB - Until recently it was thought that children do not suffer as much from postoperative pain as adults. Coupled with the fear of administering systemic opiates to young children, this meant that babies were often left "to cry it out." Lately it has been acknowledged that children and even babies not only feel pain, but similar to adults, suffer from the physiologic consequences of the untreated stress response. It has also been shown that preventing this response improves the postoperative period in babies after undergoing cardiac surgery. Regional analgesia is commonly used to provide postoperative pain control in adults and children. Following minor lower abdominal surgery, pain relief is often provided by caudal analgesia or specific nerve blocks. Epidural and spinal anesthesia are also gaining popularity for young children and even babies. We describe 3 cases of continuous epidural analgesia in babies, and review other methods for postoperative pain relief. These cases are representative of the many which we have treated over the past 2 years. We use epidural analgesia in children who undergo major thoracic, abdominal, pelvic and major lower limb surgery, and are expected to suffer significant postoperative pain for more than 24 hours. We have not encountered any major complications or inadvertent dural punctures. We judiciously monitor these children, especially their respiratory function, as long as they receive epidural narcotics. PMID- 8707172 TI - [Pattern of visits to a primary care clinic by recent Russian immigrants]. AB - The pattern that characterized 2790 visits to a primary care clinic by recent immigrants from the former Soviet Union during their first 2 years in Israel was studied. The control group comprised 2689 visits of veteran Israeli patients to the same primary care clinic during the same period. 2/3 of the immigrants visited the clinic in the first year of the study and 1/3 in the second year. Half the veteran Israeli patients visited the clinic in the first year and the other half in the second. The mean number of clinic consultations for the immigrants was 5.2 per patient in the first year as opposed to 3.2 in the second. In the control group there was an average of 3.8 visits in the first year, and 4.1 in the second. The commonest reasons for consultations by immigrants were circulatory diseases, and for the controls, respiratory diseases. In both years there was a significant statistical difference between the number of visits by immigrants which were associated with elevated blood pressure, angina pectoris and psychological problems, compared to veteran Israelis. Thus, most of consultations by immigrants took place in the first year for circulatory diseases. There followed a gradual decrease in consultations, indicating successful management of their medical problems and perhaps adaptation to the behavioral pattern of veteran Israelis. PMID- 8707173 TI - [Transrectal ultrasound-controlled needle aspiration of a prostatic cyst]. AB - Prostatic cysts are common and are usually acquired and asymptomatic. Congenital prostatic cysts, which may originate from the wolffian system or from remnants of the Muellerian duct, are rare and may cause obstructive symptoms in young adults. Due to the availability of transrectal ultrasonography an increased number of cases of prostatic cysts are being diagnosed. We report a 36-year-old man with a congenital prostatic cyst which caused increasing pain during ejaculation and decreased the force of the urinary stream. It was diagnosed by ultrasonography and treated successfully by ultrasound-controlled, transrectal needle aspiration. There was immediate, complete disappearance of symptoms and no complications. PMID- 8707174 TI - [Watermelon stomach: a cause of chronic blood loss in the elderly]. AB - Watermelon stomach, or gastric vascular ectasia, is a relatively new, distinct clinical entity. This rare vascular abnormality of the stomach causes severe chronic blood loss and iron-deficiency anemia, particularly in the elderly. Endoscopically it is characterized by longitudinal antral folds which contain visible vessels radiating from the pylorus, and resembling the dark stripes on the surface of some watermelons. This endoscopic picture may easily be misinterpreted as antral hemorrhagic gastritis. The specific histologic features which establish the diagnosis on endoscopic biopsy are dilation and thrombosis of mucosal capillaries and fibromuscular hyperplasia of the lamina propria. Associated diseases, including achlorhydria, pernicious anemia and chronic liver disease have been reported. We recently encountered 2 cases of watermelon stomach associated with pernicious anemia and hypothyroidism. This entity should be considered in the differential diagnosis of iron-deficiency anemia. PMID- 8707175 TI - [Gastric phytobezoar]. AB - A case of a large gastric phytobezoar (5 x 8 cm) is presented. It was diagnosed at operation in the stomach of a post vagotomy-pyloroplasty patient, and all attempts at removal (crushing, lavage and diathermy) failed. Complete removal of the phytobezoar was accomplished in 3 endoscopy sessions using a very large snare introduced through an overtube. The patient was told to refrain from cellulose- and fiber-rich food (such as persimmons and oranges) and was given prokinetic drugs, including cisapride, domperidone and metoclopramide. PMID- 8707176 TI - [Roles of the government in tobacco control]. PMID- 8707177 TI - [Interaction between natural killer (NK) cells and the major histocompatibility system]. PMID- 8707178 TI - [Therapeutic potential of ozone--possible side-effects]. PMID- 8707179 TI - [Diseases of the gallbladder and biliary tract in patients with AIDS]. PMID- 8707180 TI - [Special ocular movements in the neonate and Bell's phenomenon]. PMID- 8707181 TI - [Behind closed doors--the hidden epidemic]. PMID- 8707182 TI - [Atherosclerotic disease of the aorta: a source of thromboembolism]. PMID- 8707183 TI - [Pregnancy-associated osteoporosis]. PMID- 8707184 TI - [Overzealous evaluation of the thyroid nodule]. PMID- 8707185 TI - [Localized scleroderma in children]. PMID- 8707186 TI - [Physiologic importance of dehydroepiandrosterone]. PMID- 8707187 TI - [Findings of periodic thoracic X-ray screening of Israel Defense Forces career personnel]. AB - Findings of routine, periodic, medical examinations of approximately 5000 Israel Defense Forces (IDF) career personnel, included a screening postero-anterior chest radiograph were reviewed. Previous studies have indicated low efficacy of routine chest X-rays in various populations. We evaluated this screening test among 4593 career military personnel. Their chest X-rays were reviewed retrospectively and in 186 (4%) there were abnormal findings. The rate of pathological findings was positively correlated with age. Additional information was found in the X-rays of 25 subjects (0.54%), but the original interpretations of 51 (1.1%) were found to be false-positive. In view of the very low yield of findings in routine chest X-rays of IDF career military personnel, we recommend X ray screening only in high-risk populations and subjects over the age of 50. PMID- 8707188 TI - [Packing and abbreviated laparotomy in the injured--a life-saving procedure]. AB - Packing of abdominal viscera and abbreviated laparotomy are gaining favor among surgeons for cases with multiple injuries. During a 2-year period, 10 patients required abbreviated exploratory laparotomy for abdominal trauma. The 6 records which were complete form the basis of this study. 3 patients were hypotensive at the scene of injury and in the emergency department. Major vascular injury was encountered in 4. The only procedures attempted at the initial operation were packing bleeding solid viscera and controlling bleeding and fecal spillage. Hypothermia, acidosis and need for massive blood transfusion were indications for terminating exploration without attempting definitive surgery. PMID- 8707189 TI - [Quantitative analysis of spermatogenesis in fine-needle aspirates of the testicle]. AB - It was shown recently that testicular fine-needle aspiration (TFNA) can replace biopsy as a diagnostic tool in male infertility. The aim of this study was to determine whether a method of quantitative analysis of spermatogenesis can be developed based on the cytological elements in the aspirates. TFNA aspirates from markedly oligospermic and azoospermic patients were studied. All spermatogenic cells (spermatogonia, spermatocytes, spermatids and spermatozoa, as well as Sertoli cells) in the aspirates were identified and counted. It was found that: 500 consecutive cells were needed to reach conclusions about the spermatogenic process; the number of Sertoli cells in aspirates was constant in each individual's smears; each spermatogenic cell type could therefore be related to the number of Sertoli cells; the cell type index (CTI: the ratio between the number of cells of each given cell type and the number of Sertoli cells in the count of 500 consecutive cells) indicated the relative appearance of each cell type in the spermatogenic process. Determination of a spermatogenic process index (SPI), reflecting the ratio of the CTI for a given spermatogenic stage to its previous one, may be used in a flow sheet of the spermatogenic process. Assessing these indices for all the spermatogenic cell types in testicular aspirates provides a method for quantitative evaluation of the process of spermatogenesis. It enables expression of testicular pathology quantitatively and it defines various testicular fertility disorders. It can also be used as a guide for surveillance of therapy. PMID- 8707190 TI - [Direct percutaneous transluminal coronary angioplasty]. AB - Early reperfusion in acute myocardial infarction (AMI) effectively reduces mortality. Direct. percutaneous. transluminal coronary angioplasty (PTCA) is a recognized alternative to intravenous thrombolytic therapy in early AMI. Of 289 PTCAs performed during a year, 29 (10%) were direct PTCAs. Indications for direct PTCA in 21 patients (72%) were large infarcts, and in 8 (28%) cardiogenic shock. The left anterior descending artery was involved in 15 patients (52%), the right coronary artery in 10 (34%), the left circumflex in 3 (10%), and the left main artery in 1 (3%). Satisfactory opening of the occlusion was achieved in 26 (90%); there was partial success in 2 (7%) and failure in 1 (3%). 4 patients required stent deployment for treatment of dissection. There was no major complication. A patient in whom success was only partial, and the one in whom PTCA failed, were sent for operation. These data do not differ from our success rate in elective PTCA. It is concluded that direct PTCA is a safe procedure with a high rate of successful reperfusion in AMI. PMID- 8707192 TI - [Uncommon presentation of perforated colonic diverticuli]. AB - Diverticulosis of the colon is common in the sigmoid, but much less so in the cecum, ascending or transverse colon. We report 2 patients who presented with clinical findings of perforated diverticulitis of the transverse and left colon. A 42- and a 46-year-old man underwent extended right and left hemicolectomies, respectively. The first patient ran an excellent postoperative course. In the second, in whom a double-barrelled colostomy was formed, the proximal colostomy had to be redone because of stomal retraction, but both stomata eventually were closed. On follow-up both patients are symptom-free. The diagnosis of diverticulitis with complications should be considered in patients who present with intra-abdominal masses. Preoperatively, the condition often cannot be differentiated from carcinoma. PMID- 8707191 TI - [Progression of hepatitis C in a woman with primary immunologic deficiency]. PMID- 8707193 TI - [New infant formula based on soy milk and corn flour]. AB - In recent years the use of infant feeding formulas based on soy bean has been increasing. We report a trial of a new, soy bean-based formula to which starch was added. A control group was given a cow milk-based formula widely used in this country ("Materna" with cornflour, Ma'abaroth Products Ltd.). Healthy infants, between 3-6 months of age and of normal length and weight were enrolled at random at Infant Welfare Centers in Jerusalem. 25 received the soy formula, and 21 the cow milk-based formula. During the 2-month trial the infants did not receive any other foods. They were examined by the senior author every 2 weeks and weight, length and head circumference were measured at their Infant Welfare Clinic visits. The mothers filled questionnaires concerning the amount of formula ingested, the nature and number of stools, and complaints, such as crying. Both groups gained weight at the normal rate during the trial. In each group, the mothers were satisfied with the product and wanted to continue with it after the trial. We conclude that a soy-based formula containing starch is comparable to a cow milk formula containing star. PMID- 8707194 TI - [Training accreditation and audit in laparoscopic surgery]. PMID- 8707195 TI - [Fungi and autoimmune conditions]. PMID- 8707196 TI - ["Is he who refuses to enlist in the army a patient?" Psychiatrists' dilemmas during war and peace in the absence of consensus]. PMID- 8707197 TI - [Preventive medicine--hypercholesterolemia and the family physician's role]. PMID- 8707198 TI - [Indications for hysterectomy--a time for re-evaluation]. PMID- 8707199 TI - [Treatment when blood pressure rises or falls in hypertension--new considerations for optimal prevention of cardiovascular damage]. PMID- 8707200 TI - [Japanese encephalitis--implications for travelers to the Far East]. PMID- 8707201 TI - [Are the concepts of dementia according to DSM-4 and ICD-10 similar? Some theoretical aspects]. PMID- 8707202 TI - [Analytic review in the era of the scientific information explosion]. PMID- 8707203 TI - [Capsaicin for arthritic pain]. PMID- 8707204 TI - [Misplaced music, or against "Foggy Furious". Concerning the article "He who refuses to enlist in the army: is he a patient? Psychiatrists' dilemmas during war and peace in the absence of consensus"]. PMID- 8707205 TI - [Ethical commitment during mass casualties]. PMID- 8707206 TI - [Scoop-and-run versus stabilize-and-transfer]. PMID- 8707207 TI - [Transthoracic endoscopic sympathectomy--anesthetic aspects]. PMID- 8707208 TI - [Multiple "incidentalomas"]. PMID- 8707209 TI - [Genetic counseling for families at high cancer risk]. AB - About 5-10% of the most common cancers, such as breast, colon and melanoma, result from mutations in inherited predisposition genes. Recently some of these genes have been mapped or even cloned. These advances in cancer genetics have made more precise genetic counseling possible for cancer patients and their families. In our clinic for specific genetic counseling 180 families with a history of cancer were seen during a 10-month period. In counseling sessions, the family history was confirmed and interpreted, personal risk was estimated and the availability of molecular genetic testing was presented. Blood samples for DNA testing were drawn from those with certain criteria who wished to be tested. Instructions for early detection were also given, depending on the personal risk of cancer as compared to that of the general population. PMID- 8707210 TI - [Conservative approach to blunt hepatic and splenic trauma in childhood- experiences in the Pediatric Surgery Department, Soroka Hospital in the years 1980-1994]. AB - 42 children with blunt hepatic and splenic trauma, seen from 1980-1994, were treated conservatively, but 15 with splenic and 2 with hepatic trauma underwent surgery (7 had splenic preservation procedures: partial splenectomy or splenorrhaphy). Boys predominated, the mean age was 8.6 years and the main cause of injury was road accidents. In most cases hemodynamic instability within the first 24 hours was the most significant factor in determining the appropriate approach. Those who were hemodynamically stable after this period could be safely managed without operation. No complications were attributed to either operative or conservative treatment. In 5 cases there were mild respiratory or urinary tract infections unrelated to the initial trauma; there were no deaths. The overall results support the prevailing conservative approach to the treatment of blunt splenic and hepatic trauma in the hemodynamically stable child. CAT is our preferred diagnostic imaging modality in the acute phase, and ultrasonography in the follow-up period. PMID- 8707211 TI - [Ambulatory liver biopsy]. AB - During 1992-1994 we performed 77 needle liver biopsies on ambulatory patients. 59 patients were observed for 6 hours and then released. 15 who lived more than 40 km away and 3 in whom complications were feared were hospitalized. All were released within 24 hours and there were no complications. We conclude that ambulatory needle biopsy of the liver is safe and cost-effective is selected patients. PMID- 8707212 TI - [Prolonged fever in painless subacute thyroiditis--diagnosis by gallium scan]. AB - Patients with subacute, granulomatous thyroiditis usually present with a typical clinical picture of fever, anterior neck pain and an exquisitely tender thyroid. Abnormal thyroid function tests and suppressed radio-iodine uptake strongly support the diagnosis. Occasionally the disease may simulate systemic or malignant disease when it presents atypically, without signs or symptoms directly related to the thyroid, We report a 43-year-old man who presented with fever of unknown origin. A diagnosis of silent thyroiditis was made from evidence of biochemical thyrotoxicosis, intense gallium-67 citrate thyroidal localization, and an excellent response to aspirin. PMID- 8707213 TI - [Laparoscopic cholecystectomy in acute cholecystitis]. AB - We performed 417 laparoscopic cholecystectomies, including 58 for acute cholecystitis, between September 1991 and April 1995,. All operations were successful, with no mortality or complications. In about 10%, the laparoscopic approach failed and we converted to open cholecystectomy. Average post-operative hospitalization was 24 hours. We also performed primary open cholecystectomies in 55 patients with acute cholecystitis, because of limitations of operating room and staff availability for unscheduled laparoscopic surgery. In these patients, hospital stay was longer and rate of complications higher. In our opinion laparoscopic cholecystectomy is safe and the preferred approach in acute cholecystitis. PMID- 8707214 TI - [Optic nerve sheath enlargement and reversal of optic nerve head in pseudotumor cerebri]. AB - Using standard cerebral computerized tomography (CT), we diagnosed pseudotumor cerebri (PTC) and correlated the CT findings with CSF pressure and severity of visual impairment. 13 patients with a clinical diagnosis of PTC were compared with 20 age-matched controls with headache, but without papilledema or other neurologic signs. Cerebral CT consisted of axial sections of the posterior fossa, including the orbits. In all subjects the diameter of the optic nerve sheath, reversal of the optic nerve head, presence of empty sella, and size of the ventricles, cisterns and sulci were evaluated. There were no differences in basal cisterns and ventricles between those with PTC and control subjects. Empty sella was found in 6 of 13 PTC patients, compared with 1 of the 20 controls. Optic nerve sheath diameter in controls ranged from 3.5-5.0 mm (average 4.2 +/- 0.54 mm) but from 4.5-9.0 mm (average 6.8 +/- 1.54 mm) in those with PTC. Reversal of the optic nerve head was seen in 4 cases of PTC but in none of the controls. In PTC patients with opening CSF pressure greater than 270 mm water, the diameter of the optic nerve was wider than 7.5 mm. Thus, in most cases of PTC, bilateral enlarged optic nerves can be measured by standard cerebral CT and intracranial space-occupying lesions can be excluded as well. Moreover, reversal of optic nerve head, and empty sella can frequently be seen on CT in those with PTC. PMID- 8707215 TI - [Nevoid basal-cell carcinoma syndrome in a patient with cleft lip and palate]. AB - In a 48-year-old man with the nevoid basal-cell carcinoma, Gorlin-Goltz syndrome was diagnosed. There were multiple, facial, basal-cell carcinomas, multiple, odontogenic keratocysts of the mandible, rib malformations, and a wide nasal bridge, common characteristics of the syndrome. Cleft lip and palate, seen in this patient, has been described in only 5% of those with this syndrome. PMID- 8707216 TI - [Whiplash injury--a valid diagnosis]. PMID- 8707217 TI - [New treatments in HIV]. PMID- 8707218 TI - [Electro-convulsive therapy--review of law and literature]. PMID- 8707219 TI - [Transport of critically ill patients]. PMID- 8707220 TI - [Significance of skin surface pH]. PMID- 8707221 TI - [Dopamine and the kidney--renal sparing effect: fact or fiction?]. PMID- 8707223 TI - [Sphincter of Oddi dysfunction]. PMID- 8707222 TI - [The disabled in the community--towards equal opportunity]. PMID- 8707224 TI - [Use of topical anesthesia--EMLA cream]. PMID- 8707225 TI - [Pre-hospital multicasualty management]. PMID- 8707226 TI - [Nonoperative treatment of blunt splenic injuries in adults]. PMID- 8707227 TI - [Anti-sense in medicine and central nervous system disorders]. PMID- 8707228 TI - Karyotype polymorphism in the slender loris (Loris tardigradus). AB - Three wild-caught Sri Lankan and two captive-bred slender lorises (Loris tardigradus) in Adelaide Zoo, South Australia, were karyotyped from leucocyte cultures. The founder male and one of the founder females were of the grey subspecies L. tardigradus grandis, of which L. tardigradus nordicus is probably a synonym. The other founder female was a small red-brown L. tardigradus tardigradus. All three wild-caught individuals were karyotypically distinct from each other and their offspring showed the expected heterozygosity. The L. tardigradus tardigradus karyotype was no more distinct from the grey subspecies karyotypes than they were from each other. On present evidence, Indian and Sri Lanka lorises appear to be chromosomally distinct. PMID- 8707229 TI - Periodicity of calling by the greater bushbaby (Galago garnettii) on Pemba Island, Zanzibar. PMID- 8707230 TI - Feeding on a marsh-living herbaceous plant by black howler monkeys (Alouatta caraya) in central Brazil. PMID- 8707231 TI - Field observation of twinning in the dusky titi monkey, Callicebus cupreus. PMID- 8707232 TI - The anterior dentition of adapids and anthropoid origins. AB - A recent discussion of the anterior teeth of adapids led to the conclusion that similarities to anthropoids are convergent. It is shown here that alternative functional and phylogenetic interpretations can be supported. These alternative interpretations are based on a different perspective on the anterior dentitions of extant and fossil anthropoids, which cannot be characterized by a single pattern of morphology and occlusion. More importantly, when this interspecific variation is placed within a phylogenetic framework it reveals persistent evolutionary trends that can be extrapolated back to infer a modest range of probable conditions in an Eocene anthropoid ancestor. The morphology and occlusion of adapid anterior teeth fall within this extrapolated range. This is compatible with an adapid ancestry for anthropoids. PMID- 8707233 TI - Foraging and ranging patterns in white-bellied spider monkeys. AB - Ranging and foraging patterns observed during a field study of the behavioral ecology of Ateles belzebuth belzebuth are analysed in the context of temporal and spatial variation in the distribution of plant resources at the study site. The study group exhibited two basic patterns of foraging behaviour and habitat exploitation, which appeared to correlate with the observed variation in resource availability. During the dry season, when the relative abundance of fruit decreased, the spider monkeys ranged over a smaller area, repeatedly visiting a few intensively used fruiting and sleeping trees. As fruit abundance increased during the course of the wet season, the study group expanded its range, and both the daily path length and the number of fruiting trees visited increased daily. These patterns are analysed and evaluated in relation to foraging strategies. PMID- 8707234 TI - [Effect of exogenous toxins on fertility]. AB - The influence of certain harmful noxae on male fertility was investigated statistically in a large group of andrological patients. A significant correlation was found to exist between alcohol consumption and potency disorders. Patients admitting to a high level of drinking also indicated greater stress at work, and an elevated use of drugs, while more tended to use the sauna. In the case of nicotine a significant influence on sperm morphology and sperm count was shown. PMID- 8707235 TI - [Environmental factors on male fertility]. AB - The fact that the incidence of infertile marriages is on the increase throughout the world has stimulated a greater interest in identifying possible effects of the environment on the fertility of both man and woman. Over the last 50 years, the average number of spermatozoa in the ejaculate has decreased by about a half. The present review takes a look at recognized noxae and their effect on fertility, with the aim of providing clinically active physicians with up-to-date information about the major noxious substances, thus enabling them to be avoided. PMID- 8707236 TI - [Allogeneic stem cell transplantation. A valuable alternative after myeloablative chemotherapy]. PMID- 8707237 TI - [Disorders of lipid metabolism--diagnosis and therapy in general practice. 4: Therapy of hypercholesterolemia--drug treatment]. PMID- 8707238 TI - [Dynamics of the hypnotic effect of 0.125 mg and 0.250 mg brotizolam. The lower dosage is adequate for treatment of sleep disorders]. AB - BACKGROUND: Although observations have shown that half a tablet (0.125 mg) of brotizolam develops an adequate sedative effect in patients who cannot get to sleep, confirmatory pharmacodynamic studies are lacking. METHOD: In a comparative double-blind, randomized, placebo-controlled study, the hypnotic effects of 0.125 mg and 0.250 mg brotizolam, a short-acting benzodiazepine-like hetrazepine, were investigated. The concept underlying the study was that both doses are equally rapid in action onset, but differ in duration of effect. The study included 49 healthy women aged between 23 and 44 years. The nocturnal tests lasted until 2.30 am, and a concluding examination took place after breakfast. The test subjects were asked to estimate their state of well-being on visual analog scales at half hour intervals, while concentration was tested hourly using the digit symbol substitution test (DSST) tried and tested in numerous studies on benzodiazepines. RESULTS: Both doses proved to be equally effective after 0.5, 1.5 and 2.5 hours. While the hypnotic effect of the lower dose no longer differed from that of placebo after 3.5 hours, the larger dose still had a greater effect than both placebo and the lower dose after 4.5 hours. A hangover effect in the morning was not observed with either dose. The pharmacodynamic results indicate that the effect of the substance is readily controllable, and that half a tablet of brotizolam is an effective treatment of difficulty in getting to sleep initially or after waking prematurely. CONCLUSION: The results of the present study demonstrate for the first time that a half-tablet of brotizolam is an effective treatment of difficulty in getting to sleep initially or after premature wakening. PMID- 8707239 TI - [Decrease in sperm count does not yet signify infertility. Interview by Elisabeth B. Moosmann, M.A..]. PMID- 8707240 TI - [Basedow disease--thyrostatic drugs combined with thyroxine?]. PMID- 8707241 TI - A typical courier day in 1940's. PMID- 8707242 TI - Student advising at a distance. PMID- 8707243 TI - Letter from a former courier. PMID- 8707244 TI - Mary Breckinridge Healthcare Services 1995 version. PMID- 8707245 TI - Hepatitis B virus infection of tupaia hepatocytes in vitro and in vivo. AB - For the systematic analysis of various clinical and molecular aspects of hepatitis B virus (HBV) infection, an experimental small animal system of HBV infection would be a great advance. The susceptibility to HBV infection, therefore, of hepatocytes from the tree shrew species tupaia belangeri was studied in vitro and in vivo. Primary hepatocytes isolated from livers of tupaias can be reproducibly infected with HBV. In vitro infection results in viral DNA and RNA synthesis in hepatocytes and secretion hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) into culture medium. Tupaias can also be infected with HBV in vivo, resulting in viral DNA replication and gene expression in tupaia livers. Similar to acute, self-limited hepatitis B in humans HBsAg is rapidly cleared from serum, followed by seroconversion to anti-HBe and anti-HBs. These data clearly tht HBV is infectious to tupaia hepatocytes in vitro and transiently in vivo. Tupaias, therefore, may become a useful model for the experimental analysis of various molecular and clinical aspects of HBV infection, including the significance of HBV quasispecies, the steps involved in hepatocarcinogenesis as well as the evaluation of various antiviral strategies. PMID- 8707246 TI - Prolonged fecal excretion of hepatitis A virus in adult patients with hepatitis A as determined by polymerase chain reaction. AB - In hepatitis A virus (HAV) infection, fecal excretion of the virus has been reported to cease shortly after symptoms occur. Although there have been several reports on detection of HAV in feces using polymerase chain reaction (PCR), the duration of fecal HAV shedding in human adult hepatitis A has not been well described. In the present study, we applied the reverse-transcription (RT)-PCR system to the detection of fecal HAV RNA in 10 patients with sporadic hepatitis A. The viral genomic RNA was detected in the stools from five patients after the onset of clinical symptoms. All stool samples collected within 10 days of onset of illness were HAV-RNA-positive, and the duration of positivity lasted from a few days to as long as 3 months. In four patients, HAV RNA was detected in the stool even after the serum alanine transaminase (ALT) levels had peaked, and in one patient well after ALT levels fell to normal. These results show that fecal shedding of HAV can last for months after resolution of symptoms and such patients could be a source of further spreading of the virus in the community. PMID- 8707247 TI - The metabolic organization of the adult human liver: a comparative study of normal, fibrotic, and cirrhotic liver tissue. AB - Little is known about the alterations of metabolic organization of the human liver tissue in chronic liver diseases. We therefore compared the distribution of the following zonal metabolic markers in 10 samples of normal liver tissue, 10 samples of fibrotic tissue, and 22 samples of cirrhotic tissue: (a) the enzymatic activities of glucose-6-phosphatase (G6P), lactate dehydrogenase (LDH), succinate dehydrogenase (SDH), nicotinamide-adenine-dinucleotide-phosphate [NAPH] dehydrogenase (ND), beta-hydroxybutyrate dehydrogenase (HBDH), and glutamate dehydrogenase (GDH); (b) the protein glutamine synthetase (GLS); and (c) albumin messenger RNA (mRNA). The normal human hepatic lobule was characterized by the periportal predominance of G6P and SDH enzymatic activities and albumin mRNAs, the perivenous predominance of ND and GDH, the restriction of GLS to a small perivenous compartment, and the predominanc of beta-HBDH at the contact of both portal tracts and centrilobular veins. In fibrosis, the overall metabolic organization of the normal liver tissue was retained. The expression of periportal markers predominated around enlarged portal tracts and that of perivenous markers around residual centrilobular veins. GLS was constantly detected at the contact of centrilobular veins. In cirrhotic nodules, no zonation was observed for most enzymatic activities or for albumin. Only G6P usually predominated at the periphery of the nodules. GLS was constantly undetectable. No difference accordingly to the etiology of the underlying disease was observed. In conclusion, the normal human hepatic lobule presents a marked metabolic zonation, preserved in fibrotic lesions, but lost in cirrhotic nodules. The alterations of the metabolic organization observed in cirrhosis might contribute to the pathogenesis of some of the metabolic disorders associated with advanced liver disease. PMID- 8707248 TI - Noninvasive assessment of hepatic triglyceride content in humans with 13C nuclear magnetic resonance spectroscopy. AB - Hepatic lipid content was assessed noninvasively in 15 patients with hepatic steatosis by 13C nuclear magnetic resonance (NMR) spectroscopy, and compared in a double-blind fashion with histological grading and morphometric quantitation of fat in liver biopsies taken within 2 weeks of the study. The lipid content in the liver biopsies was expressed as the volume fraction of total parenchyma occupied by fat. Hepatic triglyceride content was determined by comparing the 13C NMR signal intensity in vivo with the signal intensity obtained from a lipid phantom of known concentrations. There was an approximately 30-fold increase in the 13C NMR signals of the saturated carbons (methyl/methylene [CH2]n) region of hepatic triglycerides from patients with grade 4 steatosis compared with those with grade 0, yielding a good dynamic range for measuring hepatic triglyceride content. The correlation coefficient between the morphometric and 13C NMR techniques was 0.89 (P < .01). These studies demonstrate that 13C NMR spectroscopy can be used to noninvasively assess hepatic triglyceride content in humans. This method may be clinically useful for diagnosis and follow-up of patients with hepatic steatosis. PMID- 8707249 TI - Increased levels of soluble complement receptor 1 in serum patients with liver diseases. AB - Complement receptor type 1 (CR1) is an integral membrane protein of many hematopoietic cells and is found in a soluble form in plasma. Preliminary data have indicated that soluble complement receptor 1 (sCR1) levels in serum were increased in patients with cirrhosis. In this study, sCR1 was measured in patients with various liver diseases with a newly established enzyme-linked immunosorbent assay (ELISA). sCR1 level was elevated in chronic active hepatitis C (24 patients, 62.6 +/- 31 ng/ML; 31 normal controls, 31.4 +/- 7.8 ng/mL, P < .001), and in cirrhosis (35 patients, 143.7 +/- 61 ng/mL, P < .001). The levels increased transiently in 3 patients who had amanita phalloides intoxication. In 25 patients with advanced cirrhosis (pretransplantation screening), there were significant inverse correlations between sCR1 and both the prothrombin index (rs = -.60, P < .002) and the aminopyrine breath test (rs = -.51, P < .01). Following liver transplantation, the levels dropped from 166 +/- 70 to 49 +/- 24 ng/mL (P < .0001), and serial measurements in the posttransplantation period showed a correlation with liver dysfunction, regardless of etiology. Since CR1 is not produced by hepatocytes, the most likely explanation for the increased level of sCR1 is reduced is reduced catabolism. Thus, sCR1 may be added to the short list of large glycoproteins that accumulate in liver disease. PMID- 8707250 TI - Serum concentrations of 7alpha-hydroxy-4-cholesten-3-one reflect bile acid synthesis in humans. AB - Serum concentrations of 7alpha-hydroxy-4-cholesten-3-one (alpha-HC) have recently been shown to reflect the activity of cholesterol 7 alpha-hydroxylase in humans. To evaluate the relationship between alpha-HC in serum and bile acid synthesis, serum concentrations of alpha-HC, and rates of bile acid synthesis, as measured by the isotope dilution technique using gas chromatography-mass spectrometry, were determined simultaneously. Regression analysis revealed a positive linear correlation of serum alpha-Hc with synthesis of cholic acid (CA) (r = .59, P = .02), chenodeoxycholic acid (CDCA) (r = .75, P = .001), and total synthesis of both primary bile acids (r = .83, P < .001) in patients with gallstones and normal liver function (n = 15). alpha-HC was also correlated to input rates of deoxycholic acid (DCA) (r = .53, P <.05). Addition of patients with chronic cholestatic liver disease (n = 5) improved the correlation between serum alpha-HC and synthesis of CA (r = .75, P < .001), CDCA (r = .77, P < .001), and both primary bile acid combined (r = .87), P < .001). Our data are in agreement with the concept that synthesis of bile acids is regulated by cholesterol 7 alpha hydroxylase activity and that alpha-HC in serum may serve as a convenient marker for the semiquantitative assessment of bile acid synthesis in humans. PMID- 8707251 TI - High insulin-like growth factor binding protein 1 levels in cirrhosis: link with insulin resistance. AB - Hyperinsulinemic euglycemic clamps were performed on six patients with compensated alcoholic cirrhosis and on six normal comparison subjects. As in previous studies, glucose uptake in the cirrhotic patients was only 21% of the comparison value. The cirrhotic patients had high growth hormone (GH) and low insulin-like growth factor-I (IGF-I) levels, with low insulin-like growth factor binding protein (IGFBP)-3 levels, but surprisingly high IGFBP-I levels (26.8 +/- 8.4 microgH vs. 3.2 +/- 0.2 microm/L, P < .001). The log IGFBP-1 level was inversely correlated with the log insulin sensitivity (r = -.95). The clamps were repeated with a somatostatin infusion to suppress GH secretion. IGFBP-1 increased in both groups, especially in the cirrhotic subjects. Insulin sensitivity increased in the normal subjects but was unchanged in the cirrhotic patients. Following GH treatment (0.13 U/kg/d for 5 days), the clamps were repeated. GH, IGF-1, and IGFBP-3 levels were now similar in the two groups; IGFBP-1 levels decreased in the cirrhotic patients but remained fivefold higher than the comparison value (10.6 +/- 3.7 vs. 2.1 +/- 0.4, P < .05). Glucose uptake in the cirrhotic patients remained only 29% of the comparison value, but the change in their insulin sensitivity was inversely correlated with the change in their IGFB 1 levels (r = -.84). These results suggests an important role for IGFBP-1 in modulating insulin sensitivity in cirrhosis. PMID- 8707252 TI - S-adenosyl-L-methionine prevents disruption of canalicular function and pericanalicular cytoskeleton integrity caused by cyclosporin A in isolated rat hepatocyte couplets. AB - Isolated rat hepatocyte couplets were used to study the effect of S-adenosyl-L methionine (SAMe) treatment on disruption of canalicular function caused by cyclosporin A (CyA). Canalicular function was assessed by counting the percentage of couplets that were able to accumulate the fluorescent cholephile choly-lysyl fluorescein (CLF) into the canalicular vacuole between the two cells, i.e., canalicular vacuole accumulation (CVA). Cotreatment with 1 mmol/L SAMe prevented the inhibition of canalicular vacuole accumulation caused by CyA (75 nmol/L and 100 nmol/L), whereas treatment with it after CyA was unsuccessful. SAMe prevented the dose dependent reduction caused by CyA (5 nmol/L-1 mumol/L) both on CVA and on retention of CLF preaccumulated within the canaliculus, the effect on retention being complete. No difference in intracellular content of reduced glutathione (GSH) between the control and any dose level of the immunosuppressor, with or without SAMe treatment was observed, suggesting that changes in intracellular reduced GSH levels are not involved in the effects of SAMe. F-actin was stained with fluorescein-isothiocyanate phalloidin and fluorescence measurements were performed by confocal microscopy. The ratio of the percanalicular area fluorescence/total couplet fluorescence, indicative of F actin distribution, significantly decreased with CyA. However, cotreatment of CyA with SAMe protected the integrity of the pericanalicular cytoskeleton, suggesting that this beneficial effect on canalicular function may maintain canalicular contractions and/or preserve tight junction function. Results are discussed in relation to possible involvement of the transmethylation pathway, modifications in membrane fluidity, effects on bile acid transport, and of inhibition of uptake of CyA. They suggest that SAMe could be a good candidate for protecting against CyA-induced membrane dysfunction. PMID- 8707254 TI - Role in nitric oxide in Kupffer cell-mediated hepatoma cell cytotoxicity in vitro and ex vivo. AB - The metabolic change in tumor cells (AH70, a rat hepatoma cell line) cocultured with isolated rat Kupffer cells were visualized and analyzed by a laser scanning confocal imaging system. When AH70 cells were cocultured with Kupffer cells, fluorescence intensity of rhodamine 123 (Rh123) decreased, indicating the reduction of mitochondrial function. The reduction in Rh123 was eliminated by NG monomethyl-L-arginine (L-NMMA), an analogue of L-arginine, suggesting the involvement of nitric oxide (NO). Two hour after the cells were cocultured, membrane compromised AH70 cells which were observed as propidium from 2.8% to 25%. This increase was also attenuated by L-NMMA, suggesting that Kupffer cell mediated injury of tumor cells largely depends on NO. The concentrations of NO-2 + NO-3 in the culture medium markedly increased after coculture of AH70 cells with Kupffer cells. Moreover, NO synthase (NOS) activity in Kupffer cells significantly increased after coculture. These in vitro results suggest that NO mediates Kupffer cell-induced tumor cell damage characterized by reduced mitochondrial function and diminished barrier function. In the ex vivo study of the perfused liver to which AH70 cells were injected via the catheter inserted into the portal vein, some AH70 cells were arrested in the upper stream of sinusoid and the fluorescence intensity of Rh123 in adherent AH70 cells decreased in a time-dependent manner within 2 hours. The number of PI-positive AH70 cells also increased 2 hours after the injection of AH70 cells. These changes were inhibited by either administration of N omega-L-nitroarginine-methylester (L NAME) to perfusate or pretreatment of the rat liver with GdCl3, which is known to deplete Kupffer cell function. Thus, the present study suggests that NO from Kupffer cells induces mitochondrial dysfunction in tumor cells followed by membrane barrier dysfunction in the liver sinusoid. PMID- 8707253 TI - Expression and immune response to hepatitis C virus core DNA-based vaccine constructs. AB - Hepatitis C virus (HCV) is a major worldwide cause of acute and chronic hepatitis, cirrhosis, and hepatocellular carcinoma. The development of vaccines against HCV have been complicated by the high variability of the envelope region, and it is likely that the cellular immune responses to viral structural proteins may be important for eradicating persistent viral infection. Recently, it was reported that the injection into muscle cells of plasmids encoding viral genes resulted in the generation of strong cellular immune responses. We constructed vectors that express the highly conserved HCV core gene. In this regard, the pHCV 2-2 construct contained the entire HCV core region and pHCV 4-2 contained both the 5' noncoding region and the core gene. Cellular expression of HCV core protein was assessed following transfection into human and murine cell lines, and higher intracellular levels of the 21-kd core protein were observed with pHCV 2 2. These HCV core DNA constructs were used to immunize BALB/c mice and produced low-level anti-HCV core humoral immune responses. To assess cytotoxic T lymphocyte (CTL) activity generated in vivo, a cloned syngeneic SP2/O myeloma cell line constitutively expressing HCV core protein was established and inoculated into BALB/c mice to produce growth of plasmacytomas. Strong CTL activity was generated because the tumor size and weight in pHCV 2-2-immunized mice were remarkably reduced compared with mice injected with mock DNA. Spontaneous CTL activity was also exhibited by splenocytes in an in vitro cytotoxicity assay. These investigations demonstrate that plasmid constructs expressing HCV core protein generate strong CTL activity, as assessed both in vivo and in vitro, and are promising candidates as antiviral agents. PMID- 8707255 TI - Effects of polyamine imbalance on the induction of stress genes in hepatocarcinoma cells exposed to heat shock. AB - The expression of hsp70-the inducible member of the corresponding heat shock gene family-of the oxidative stress marker gene heme oxygenase (HOx), and of the immediate early response genes c-fos and c-jun has been studied in FAO hepatocarcinoma cells depleted of polyamines and exposed to heat shock. Depletion of polyamines was obtained in short-term experiments (24-48 hours) by the use of alpha difluoromethylornithine (DFMO), a classical inhibitor of ornithine decarboxylase (ODC), or of the combination of the newly available inhibitors of ODC and S-adenosylmethionine decarboxylase, i.e., (2R,5R)-hept-6-yne-2,5-diamine (MAP) and 5'{[(Z)-4-aminobut-2-enyl]methylanino}-5-deoxyadeno-si ne (AbeAdo). Under our experimental conditions polyamine imbalance was realized without appreciable growth-related genes. Decreases of putrescine and spermidine 48 hours after DFMO prevented the induction of hsp70 messenger RNA (mRNA), whereas depletion spermidine and spermine obtained with MAP/AbeAdo decreased intensity and duration of post-heat shock accumulation of hsp70 mRNA. Inductions of HOx, c jun and c-fos were also inhibited. Because MAP/AbeAdo caused also an intracelluar accumulation of putrescine, we tested the effect of exogenous putrescine, which was found to stabilize the mRNAs for hsp70 and c-jun. Hsp70 and HOx are thought to play a protective role, and the proteins of c-jun and c-fos constitute the transcription factor activator protein-1, which is involved in the transcription of many defensive products. Therefore, the integrity of polyamine pool seems to be a necessary permissive condition for an effective response of the cells to adverse environmental changes. PMID- 8707256 TI - Glycohydrolases as markers of hepatic ischemia-reperfusion injury and recovery. AB - Advances in liver surgery and transplantation have lead to a steady increase in the number of these interventions. Prompt quantitative assessment of hepatic of hepatic function and a patient's subsequent morbidity and mortality following surgery remain difficult despite the currently utilized historic markers of hepatic parenchymal injury (e.g., aspartate transaminase [AST], lactate dehydrogenase [LDH] gamma-glutamyl transpeptidase [GGT]). Increases in serum glycohydrolase activities appear to provide sensitive and quantitative markers of hepatic ischemia/reperfusion injury. In 10 male swine (25 to 35 kg body weight) following 30, 45, and 90 minutes of acute hepatic ischemia, the systemic release of eight different glycohydrolases and lipid peroxides into serum were determined and compared with pre- and postischemic serum levels of LDH, GGT, and AST. The rapid release of glycohydrolases into serum was directly proportional to the length of the ischemic period from 30 to 90 minutes; e.g., beta-glucosidase, mean 1.9-fold increase at 30 minutes; 8.3-fold at 45 minutes; and 22.8-fold at 90 minutes; P < .002) and the activities peaked within the first 3 hours postischemia. In constrast, AST, LDH, and GGT were released slowly and peaked 20 to 30 hours after hepatic blood flow was restored. In swine with fatal outcomes (90 minutes of ischemia), all enzyme levels increased continuously during the final hours of life. However, in swine that survived hepatic ischemia/reperfusion injury (45 minutes of ischemia) the glycohydrolases, but not AST, LDH, and GGT, declined after 2 to 3 hours' postischemia and the serum lipid peroxide levels followed the same pattern. Serum beta-galactosidase and beta-glucosidase levels are sensitive markers that rise as quickly as traditional enzyme markers (AST, LDH, GGT) following hepatic ischemic injury; moreover, the glycohydrolases have the added value of serving as predictors of survival. PMID- 8707257 TI - L-arginine minimizes reperfusion injury in a low-flow, reflow model of liver perfusion. AB - A low-flow, reflow model of liver perfusion was used in the rat to investigate the effects of L-arginine on reperfusion injury in the absence of blood elements. In contrast to in vivo studies, L-arginine cannot minimize hypoxia by improving the microcirculation under these special conditions, but rather can only increase oxygen delivery upon reflow. During reflow, lactate dehydrogenase (LDH) release reached a new steady-state value of 35 +/- 3 U/g/h in livers perfused in the absence of L-arginine. L-Arginine (0.1 mmol/L) significantly reduced LDH release during reflow to 14 +/- 1 U/g/h; higher concentrations (1.0-3.0 mmol/L) were less effective and the arginine analogue Nomega-nitro L-arginine methyl ester (L-NAME, 0.3 mmol/L) reversed the protective effect completely. Infusion of the biologically inactive isomer D-arginine (0.1 mmol/L had no effect on the observed reperfusion injury. Malondialdehyde (MDA) release during reflow averaged 92 +/- 10 nmol/g/h and was decreased significantly to 47 +/- 13 nmol/g/h with L-arginine (0.1 mmol/L). Oxygen uptake during reflow was not significantly altered by L arginine, although the time required to reach steady state values upon reflow was reduced significantly (about twofold), indicating improved microcirculation. Trypan blue distribution time, which is used to index the hepatic microcirculation, was decreased significantly from 330 +/- 17 to 227 +/- 31 seconds by L-arginine, an effect also blocked by L-NAME. Additionally, L arginine, significantly increased both the rate of entry and exit of fluorescein dextran, a dye confined to the vascular space, by approximately 50%, also reflecting improved microcirculation. Collectively , these findings indicate that L-arginine protects against hypoxia/reoxygenation injury in a blood-free perfusion model specifically during the reoxygenation period. It is likely that L arginine rapidly removes substrates for free radical generation by improving the microcirculation. PMID- 8707258 TI - Liver cell uptake and degradation of soluble immunoglobulin G immune complexes in vivo and in vitro in rats. AB - Immune complexes were formed between dinitrophenylated human serum albumin (DNP HSA) and polyclonal rabbit immunoglobulin G (IgG) anti-DNP antibodies at antibody excess. The antigen was labelled with isotope (125I-tyramine-cellobiose) or fluorochrome, (6-[fluorescein-5-(and-6)-carboxamido] hexanoic-acid, succinimidyl ester). The radiolabelled antigen, native or antibody complexed, was given intravenously to rats. Radioactivity was measured in various organs at 1 hour following injection. The liver was the main site for removal of the antigen as well as of the immune complexes. Within the liver, immune complexes were mainly associated with nonparenchymal liver cells, the total recovery from Kupffer cells being about 10 times greater than from the liver endothelial cells. The uncomplexed radiolabelled antigen was readily degraded by both cells types. After IgG complexing, the degradation decreased, both in Kupffer cells and in liver endothelial cells. In vitro experiments with isolated liver cells, showed that IgG complexing increased antigen uptake to about the same extent in Kupffer cells and in liver endothelial cells. The degradation of both antigen and immune complexes was less efficient in vitro than in vivo. Immune complex uptake in vitro was shown also by confocal fluorescence microscopy in Kupffer cells and in liver endothelial cells. Also in vitro, only minor uptake was found in the hepatocytes. We conclude that both liver endothelial cells and Kupffer cells are involved in the hepatic handling of soluble IgG immune complexes, but we found no evidence for substantial uptake by hepatocytes. PMID- 8707259 TI - Tissue inhibitor of metalloproteinase-1 messenger RNA expression is enhanced relative to interstitial collagenase messenger RNA in experimental liver injury and fibrosis. AB - Liver fibrosis results from a relative imbalance between synthesis and degradation of matrix proteins. We have previously described release of the protein collagenase inhibitor, tissue inhibitor of metalloproteinase-1 (TIMP-1), by culture-activated human hepatic stellate cells (HSCs). In this study, we have investigated the relative expression of TIMP-1 and interstitial collagenase in culture-activated rat HSCs and rat models of liver injury and fibrosis. The complementary DNA (cDNA) for rat TIMP-1 was obtained by homology polymerase chain reaction (PCR) and sequenced. By Northern analysis using this probe, TIMP-1 messenger RNA (mRNA) expression was up-regulated with HSC activation by culture on plastic as defined by cellular expression of procollagen-1. Interstitial collagenase mRNA was expressed in early 1. Interstitial collagenase mRNA was expressed in early culture (<4 days) but became undetectable in more activated cells (7-21 days). By activity assay of serum-free cell-conditioned media, TIMP-1 was found to be released in increasingly concentrations with duration of culture on plastic. Expression of TIMP-1 interstitial collagenase, and procollagen-1 mRNAs were studied in rat models of biliary and parenchymal injury (bile duct ligation and CC14 administration) by ribonuclease protein assay. TIMP-1 mRNA expression was increased at 6, 24 hours, and 3 days after bile duct ligation and was also shown to rise in acute CC14 liver injury and remain elevated as the liver became fibrotic. TIMP-1 expression preceded procollagen-1 expression in both models. In contrasts, interstitial collagenase mRNA levels remained similar to control values throughout both models of liver injury. Total cellular RNA from hepatocytes, HSCs, and kupffer cells freshly isolated from livers after acute CC14 injury was subjected to Northern analysis. TIMP-1 transcripts were observed in nonparenchymal cells only. We suggest that increased expression of TIMP-1 relative to interstitial collagenase by HSCs may promote progression of liver fibrosis in these rat models by preventing degradation of secreted collagens. PMID- 8707260 TI - Alanine protects liver from injury caused by F-galactosamine and CCl4. AB - The liver is the main organ involved in amino acid metabolism, and it utilizes glucogenic amino acids as substrates for glucose or adenosine triphosphate (ATP), but this process is impaired in clinical and experimental liver diseases. In this study, we administered high doses of amino acids in rats or cultured hepatocytes with experimental models of liver injury to examine whether such supplementation could attenuate liver damage. We found that the addition of alanine reduced enzyme leakage from primary cultured rat hepatocytes treated with D-galactosamine (D-gal), while other amino acids did not. A significant decrease of lactate dehydrogenase (LDH) leakage was observed when cells were cultured with >6 from mmol/L alanine. Alanine also reduced enzyme leakage from normal hepatocytes that were not treated with D-gal. In D-gal-treated rats, constant infusion of a high dose of alanine significantly reduced the plasma transaminase and total bilirubin levels when compared with infusion of an amino acid mixture. Bolus administration of alanine significantly prevented the elevation of plasma transaminase levels and histological liver damage in CCl4-treated rats, while fructose-1,6 bisphosphate (FDP) had little effect. Alanine might promote the restoration of damaged liver in hepatotoxicant-treated rats, because significant effect was found after the elevation in plasma transaminase levels. Alanine also prevented the decrease of cellular ATP caused by D-gal and appeared to promote ATP production in primary cultured rat hepatocytes. These results indicate that alanine reduces experimental liver damage by a direct effect on hepatocytes. PMID- 8707261 TI - Choline transport in rat liver basolateral plasma membrane vesicles. AB - Previous studies have demonstrated several pathways for the sinusoidal uptake of endogenous and exogenous organic cations, including two distinct organic cation:H+ exchanges and a separate carrier-mediated system driven by the inside negative membrane potential. In this study, the driving force(s) for the uptake of the endogenous quaternary amine, choline, were determined in rat liver basolateral plasma membrane (blLPM) vesicles. Choline uptake into an osmotically sensitive space was not stimulated by an outwardly directed H+ or inwardly directed Na+ gradient. Instead, an inside-negative K+ diffusion potential stimulated choline uptake, suggesting the presence of a conductive pathway for choline uptake in bILPM vesicles. Conductive choline uptake was confirmed by inducing variable changes in the transmembrane potential with anions of different membrane permeability. Choline uptake in blLPM vesicles exhibitied 1) temperature dependency; 2) trans-stimulation; and 3) saturability, with an approximate Michaelis constant (Km) of 0.34 mmol/L and maximum velocity (Vmax) of 0.45 nmol/mg protein/15 s. Choline uptake in blLPM vesicles was cis-inhibited by the structurally similar derivative, hemicholinium-3 and acetylcholine, but not by substrates for other organic cation transport processes identified in blLPM vesicles, including thiamine, tetraethylammonium (TEA), tri-n-butyl methylammonium (TBuMA), and N'-methylnicotinamide (NMN). These findings demonstrate an electrogenic pathway on the sinusoidal membrane for the uptake of this essential nutrient and support the existence of multiple pathways for the sinusoidal uptake of endogenous and exogenous organic cations. PMID- 8707262 TI - Expressions of basic fibroblast growth factor and its receptors and their relationship to proliferation of human hepatocellular carcinoma cell lines. AB - On six human hepatocellular carcinoma (HCC) cell lines (KIM-1, KYN-1, KYN-2, KYN 3, HAK-1A, and HAK- 1B), we examined expressions and functions of the proteins and messenger RNAs (mRNAs) of basic fibroblast growth factor (bFGF) and its receptor, i.e., fibroblast growth factor receptor-1 (FGFR-1), as well as mRNA expressions of FGFR-2 approximately 4. All six cell lines expressed the proteins and mRNAs of bFGF and FGFR-1, and at least one of FGFR-2 approximately 4 mRNAs. Two of the six cell lines (KYN-1 and KYN-3) presented significant release of bFGF in culture supernatant, while the release in the remaining four cell lines was quite small. Addition of anti-bFGF neutralizing antibody (1, 10, or 20 microg/mL) to culture medium resulted in marked suppression of cell proliferation in all cell lines except HAK-1A. On the other hand, addition of exogenous bFGF (0.1, 1, or 5 ng/mL) to culture medium stimulated cell proliferation except in KIM-1 and KYN-2. When KIM-1 was transplanted to nude mice and anti-bFGF antibody was injected subcutaneously to a space surrounding the developed tumor, tumor proliferation was significantly suppressed in nude mice that received anti-bFGF antibody than in control mice, but there were no histological differences between the groups, including blood space formation in the stroma. In conclusion, hepatocellular carcinoma (HCC) cells may possess a proliferation mechanism regulated by an autocrine mechanism, a paracrine mechanism, or both, which are mediated by bFGF/FGFR. PMID- 8707263 TI - Reconstitution of bile acid transport in the rat hepatoma McArdle RH-7777 cell line. AB - The liver recovers bile acids from the portal circulation primarily via an active process that is dependent on sodium ions. Hepatocytes lose the ability to transport bile acids in culture, and, in liver-derived permanent cell lines, this ability is severely reduced or absent. To study the importance of bile acids in regulating liver-specific functions (e.g., cellular bile acid and cholesterol metabolism), we have re-established active bile acid transport in cultured cells. The complementary DNA (cDNA) encoding the rat sodium/taurocholate cotransporting polypeptide (ntcp) was placed under the control of a cytomegalovirus promoter and transfected into the rat hepatoma cell line, McArdle RH-7777. Transfected cells were screened for the ability to take up [3H]-taurocholate. Clones that displayed the ability to take up taurocholate were expanded (designated McNtcp) and further characterized. The apparent Michaelis constant (Km) for taurocholate uptake was similar among the different clones. The observed maximum velocity (Vmax), however, differed and was positively correlated with the abundance of recombinant ntcp messenger RNA (mRNA). The highest level of taurocholate uptake activity observed in McNtcp cells was comparable with that of freshly isolated hepatocytes. Efflux of accumulated taurocholate from McNtcp cells proceeded in a manner similar to primary hepatocytes, indicating that McArdle RH-7777 cells have retained the ability to secrete bile acids. Moreover, taurocholate uptake in McNtcp cells was inhibited by other bile acid species. Based on the observed kinetic parameters, the reconstituted McArdle RH-7777 cells mimic the ability of primary hepatocytes to transport bile acids. PMID- 8707264 TI - Improved sustained response following treatment of chronic hepatitis C by gradual reduction in the interferon dose. AB - Interferon (IFN) treatment of chronic hepatitis C virus (HCV) is associated with a high rate of relapse. IFN is thought to exert its effect against HCV via direct viral inhibition and immune stimulation. We have hypothesized that relapse following termination of therapy results from the sudden withdrawal of this immune modulatory effect and that gradual reduction in the IFN dose may decrease the incidence of relapse. One hundred six patients with chronic HCV were enrolled into this 24-month controlled, randomized prospective trial. All were treated with 5 mU of interferon-alpha-2b three times a week for 6 months. Patients who achieved biochemical response were randomized to either stop or taper IFN gradually at monthly intervals as follows; 3 mu, 2 mU, 1 mU, and 0.5 mU (all three times a week). 0.5 mU twice weekly and then once weekly. Liver histology was assessed by Knodell index and HCV RNA was measured by a quantitative polymerase chain reaction (PCR) assay. Of the 92 patients who completed the initial 6 months of IFN treatment, 47 (51%) achieved biochemical response. Twenty one of these patients were randomized to stop IFN treatment and 25 to taper (1 drop-out). At randomization patients were well matched with respect to age, sex, race, serum alanine transaminase (ALT), and liver histology. Biochemical relapse was observed in 19 of 21 (91%) patients who stopped IFN treatment compared with only 60% who tapered IFN (P= .04). Virological relapse occurred in 90% of patients who stopped and only 48% of persons who tapered IFN therapy. At completion of the 24-month study patients who achieved long-term sustained biochemical response had a significantly lower mean Knodell score (3.5 vs. 6.5) and a significantly greater number were HCV RNA negative in serum (85% vs. 18%) compared with relapsers. We conclude that gradual reduction in IFN dose is associated with a significant higher rate of sustained response and clearance of HCV RNA from serum compared with abruptly stopping treatment. This in turn is associated with a significant improvement in hepatic histology supporting the premise that response to IFN therapy can prevent progression to cirrhosis. PMID- 8707265 TI - Hepatotoxicity of the herbal medicine germander: metabolic activation of its furano diterpenoids by cytochrome P450 3A Depletes cytoskeleton-associated protein thiols and forms plasma membrane blebs in rat hepatocytes. AB - Several herbal remedies have produced hepatitis in humans. The medicinal plant, germander, was recalled after its use as an adjuvant to slimming diets resulted in an epidemic of hepatitis in France. We studied the hepatotoxicity of germander in isolated rat hepatocytes. A crude fraction containing the diverse furano diterpenoids of germander, or the purified main constituents of this fraction, teucrin A and teuchamaedryn A, were hepatotoxic (correction for hepatototoxic), but not fractions containing more polar or lipophilic constituents. [3H]Teucrin A covalently bound to hepatocyte proteins. The furano diterpenoid fraction decreased cell glutathione and cytoskeleton-associated protein thiols, and led to formation of plasma membrane blebs and cell demise. Pretreatment of male rats with troleandomycin, an inhibitor of cytochrome P450 3A (CYP3A), slowed the depletion of glutathione and decreased toxicity, whereas dexamethasone, an inducer of CYP3A, had opposite effects. Female rat hepatocytes, which poorly express CYP3A, exhibited little toxicity, unless the animals were treated with dexamethasone. Feeding male rats with a sulfur amino acid-deficient diet decreased cell glutathione and enhanced toxicity, whereas supplementation of the standard diet with cystine had opposite effects. We conclude that the furano diterpenoids of germander are activated by CYP3A into electrophilic metabolites that deplete glutathione and cytoskeleton-associated protein thiols and form plasma membrane blebs. We suggest that studies in isolated hepatocytes be included in the preclinical assessment of herbal remedies. PMID- 8707266 TI - Role of Kupffer cells and the spleen in modulation of endotoxin-induced liver injury after partial hepatectomy. AB - The hypothesis that both activated Kupffer cells and the spleen may be responsible for endotoxin-induced liver injury following partial hepatectomy was investigated. Male rats were divided into a sham group receiving laparotomy alone and three groups receiving a two-thirds hepatectomy; one group was given normal saline (NS) solution as a vehicle control, one group received intravenous gadolinium chloride (GC group) (7 mg/kg body weight) for 2 days before intravenous injection of endotoxin to inhibit Kupffer cell phagocytosis, and the third group simultaneously underwent splenectomy and partial hepatectomy (SH group). As endotoxin, lipopolysaccharide (LPS) (1 mg/kg body weight) was administered intravenously 2 days after surgery. In the GC and SH groups, phagocytic activity was reduced to approximately 40% of that in the sham group. The highest plasma tumor necrosis factor alpha (TNF-alpha) level (8,544 +/- 1,223 pg/mL) was observed in the NS group at 1 hour after LPS administration, and the level was significantly reduced by GdCl3 or splenectomy (P < 0.05). Inhibition of Kupffer cell function and splenectomy attenuated functional and structural liver damage associated with the decreased hepatic infiltration of polymorphonuclear leukocytes (PMNs) and reduced priming of circulating PMNs in the early stage of endotoxemia following partial hepatectomy. Consequently, the 24-hour survival rate of the SH and GC groups was significantly improved to 50% and 80%, respectively (P < .05), while that of the NS group was 12.5%. These findings indicate that the modification of inflammatory mediator generation by splenectomy or inhibition of Kupffer cell function may be beneficial for the prevention of endotoxin-induced liver injury after partial hepatectomy. PMID- 8707267 TI - The polymeric immunoglobulin receptor is the major calmodulin-binding protein in an endosome fraction from rat liver enriched in recycling receptors. AB - Rat liver endosomes contain one major high-affinity calmodulin-binding protein (CaMBP) that now has been identified as the polymeric immunoglobulin receptor (pIgR). In isolated endosomes pIgR was enriched in the receptor-recycling compartment (RRC); lesser enrichment was found in 'early' endosome (CURL) and much less in 'late' endosome fractions (multivesicular bodies, MVB). The distribution of the major CaMBP, shown by Western blotting or by overlay with I125-calmodulin in the isolated fractions, was consistent with rapid accumulation of I125-immunoglobulin A (IgA) in RRC and CURL after intravenous injection into rats. The receptor was also found in sinusoidal plasma membranes but not in cell fractions containing apical (bile canalicular) or lateral plasma membrane domains of the hepatocyte. The interaction of pIgR with calmodulin was shown by direct binding assays and by affinity chromatography. Thus, calmodulin is the first cytoplasmic protein shown to interact with the pIgR. We postulate that calmodulin regulates pIgA trafficking in rat liver. In addition, the receptor recycling fraction emerges as an endosomal subcompartment involved in pIgA transport via pIgR. PMID- 8707268 TI - Endothelin induced contractility of stellate cells from normal and cirrhotic rat liver: implications for regulation of portal pressure and resistance. AB - Hepatic stellate cells are similar to tissue pericytes and have been shown to be contractile. In this study, we examined the effects of known mediators of stellate cell contraction on portal pressure in rat livers after carbon tetrachloride induced injury (including cirrhosis) and investigated the contractility of stellate cells as a function of liver injury. Sarafotoxin S6C, an endothelin B (ETB) receptor agonist, had minor effects on portal pressure when perfused into normal livers at concentrations known to elicit stellate cell contraction (2 nmol/L). In contrast, both endothelin-1 (2 nmol/L) and angiotensin II (8.6 nmol/L) caused a rapid and pronounced rise in portal pressure. The effects of sarafotoxin S6C (a potent stellate cell contractile agonist) on portal pressure were greater in cirrhotic than normal liver, whereas those of angiotensin II were unchanged after liver injury. Endothelin-1 and sarafotoxin S6C induced contractility of stellate cells increased in proportion to the degree of liver injury. Contractility was greatest for stellate cells isolated from cirrhotic livers, a population of cells that displayed the most activated phenotype, as measured by immunoblot of smooth muscle alpha actin. Autoradiography of cirrhotic livers after perfusion with 125I-endothelin-1 revealed binding to cells in both sinusoidal spaces and collagenous fibrotic bands, consistent with known locations of stellate cells. Finally, the mixed endothelin-A (ETA) and ETB receptor antagonist, bosentan, reduced portal pressure in portal hypertensive animals, consistent with its inhibitory effect on stellate cell contraction. We conclude that stellate cell contractility increases with progressive liver injury and is proportional to the degree of stellate cell activation, becoming most prominent in the cirrhotic liver. Endothelin-stimulated contraction of stellate cells in cirrhotic liver may contribute to increased intrahepatic resistance and portal pressure. PMID- 8707269 TI - Altered lipid metabolism in apolipoprotein E-deficient mice does not affect cholesterol balance across the liver. AB - Adaptation of cholesterol and bile acid synthesis and of biliary cholesterol secretion represent key metabolic responses to maintain cholesterol homeostasis and have been suggested to be influenced by apolipoprotein E (apoE) phenotype in humans. We have investigated hepatic metabolism and secretion of cholesterol into bile in homozygous apoE-deficient (apoE -/-) mice fed normal lab chow. Plasma cholesterol levels were 10 times higher in apoE (-/-) mice than in controls (+/+); triacylglycerol levels were only minimally affected. Hepatic cholesterol (+56%) and triacylglycerol (+232%) contents were significantly increased in apoE (-/-) mice, whereas those of cholesteryl ester and of phospholipids were similar in both groups. Lipid accumulated predominantly in periportal areas of apoE (-/-) livers. Hepatic 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG CoA reductase) messenger RNA (mRNA) level and activity were reduced by 45% and 50%, respectively, in apoE (-/-) mice. In contrast, plasma lathosterol/cholesterol ratios, indicative for whole-body cholesterol synthesis, were fourfold increased in these mice. Acyl-coenzyme A:cholesterol acyltransferase (ACAT) activity was similar in livers of both groups. Despite the marked changes in hepatic cholesterol metabolism, neither hepatic bile acid synthesis, bile acid pool size and composition, nor hepatic cholesterol 7alpha-hydroxylase and sterol 27 hydroxylase mRNA levels differed between apoE (-/-) and (+/+) mice. In addition, biliary cholesterol secretion was unaffected in the knock-out mice. Our results show that lack of apoE leads to marked changes in hepatic cholesterol metabolism without altering cholesterol balance across the liver. The data are compatible with increased peripheral cholesterol biosynthesis in apoE-deficient mice. PMID- 8707270 TI - Experimental Mallory body formation is accompanied by modulation of the expression of multidrug-resistance genes and their products. AB - Mallory bodies (MBs) are characteristic morphological features of alcoholic hepatitis and are also found in other chronic liver disorders and hepatocellular neoplasms. MBs can be produced in mouse liver by chronic administration of the porphyrinogenic drugs griseofulvin (GF) and 3,5-diethoxy-carbonyl-1,4 dihydrocollidine (DDC). The mechanisms causing the formation of MBs are poorly understood, and the significance of MB formation during the course of liver disease remains unclear. We investigated the relationship between the mechanisms underlying the formation of MBs and the regulation of multidrug resistance (mdr) genes and their products, the P-glycoproteins (Pgp). Immunofluorescence microscopy using the monoclonal antibody C219 revealed an increase of Pgp expression in almost all hepatocytes after 3 to 8 days of feeding mice DDC- and GF-containing diets. However, after approximately 4 weeks of DDC and approximately 8 weeks of GF feeding, when the first small MBs appeared and loosening and diminution of keratin intermediate filament (KIF) cytoskeleton occurred in some hepatocytes, a decrease or loss of Pgp staining in affected hepatocytes was observed. After feeding mice DDC for 6 weeks and GF for 12 weeks, many hepatocytes contained MBs and displayed a disruption of the immunohistochemically demonstrable KIF meshwork. Double immunofluorescence microscopy with the keratin polyclonal antibody and the mab C219 at this time point revealed a complete loss of Pgp staining in affected cells, although remaining hepatocytes with unaltered KIF meshwork showed a strong reaction with the C219 antibody. Northern blot analyses revealed a significant increase of mdr2 mRNA and, to a lesser extent, of mdr1a mRNA in the livers of DDC- and GF-fed animals. PMID- 8707271 TI - Alterations in glutathione homeostasis in mutant Eisai hyperbilirubinemic rats. AB - Eisai hyperbilirubinemic rats (EHBR) are mutant Sprague-Dawley rats that exhibit impaired biliary organic anion and reduced glutathione (GSH) secretion. In addition, liver GSH levels are twice that of age-matched controls. The mechanisms for the defect in biliary GSH secretion and the increase in cell GSH are not fully understood. We previously showed that canalicular membrane-enriched vesicles isolated from EHBR livers exhibited normal GSH transport. In the present study, we examined the steady-state rat canalicular reduced glutathione transporter (RcGshT) messenger RNA (mRNA) and protein levels, as well as the mechanisms for the increase in cell GSH. Both Northern and Western blot analyses of EHBR livers showed nearly identical RcGshT mRNA and polypeptide levels, respectively, as compared with controls. Treatment with phenobarbital, which increased steady-state RcGshT mRNA by five- to sixfold, RcGshT polypeptide, and biliary GSH secretion by onefold in controls, had a smaller effect on steady state RcGshT-mRNA level in EHBR (by 1.5-fold) and did not increase RcGshT polypeptide or biliary GSH secretion. In examining possible mechanisms for increased liver GSH, both cysteine level and gamma-glutamylcysteine synthetase (GCS) activity were significantly higher than controls, while the activity of GSH synthetase was unchanged. Northern and Western blot analyses also showed increased steady-state GCS heavy subunit (GCS-HS) mRNA and polypeptide levels, respectively. In addition to liver, GSH levels in kidney, duodenal, jejunal, and ileal mucosa of EHBR were 200% to 300% of age-matched control rats. GCS activity was also increased in kidney cytosol of EHBR. Thus, the defect in biliary GSH secretion in EHBR most likely is either at the posttranslational level of RcGshT or in the inhibition exerted by retained endogenous organic anions. In addition, there is a widespread up-regulation of GSH synthesis capacity in the tissues of EHBR. PMID- 8707273 TI - Redundant publication: a form of reader abuse. PMID- 8707272 TI - Second International Ringberg Conference: "Cell Biology and Molecular Basis of Liver Transport". PMID- 8707274 TI - Changes in serum fibrogenesis markers during interferon therapy for chronic hepatitis type C. AB - We serially measured the levels of serum N-terminal peptide of type III procollagen (PIIINP), the 7S domain of type IV collagen (IV-7S), and hyaluronate (HA) before (0 month), at the end (6 months), and 24 weeks after the end of interferon therapy (12 months) in patients with chronic hepatitis type C to estimate the effects of interferon alfa (IFN-alpha) on serum levels of hepatic fibrogenesis markers. One hundred twenty-one patients with chronic hepatitis type C received intramuscular injection of 6 million U of natural IFN-alpha for 24 weeks. Patients were divided into three groups: sustained complete response (CR S), complete response with rebound (CR-R), and nonresponse (NR). Serum PIIINP, IV 7S, and HA levels were significantly decreased and reached normal levels at 12 months in CR-S; only IV-7S levels were significantly decreased at 12 months in CR R, whereas these levels remained abnormally high in the NR. These results suggest that IFN-alpha therapy could lower the levels of serum hepatic fibrogenesis markers along with improvements in hepatic inflammation. PMID- 8707275 TI - Developing a World view toward acute liver failure. PMID- 8707276 TI - Redundant publication in hepatology. PMID- 8707277 TI - Magnetic resonance imaging pallidal hypersignal in cirrhotic subjects. PMID- 8707278 TI - High rate of mutations in the hepatitis B core gene during the immune clearance phase of chronic hepatitis B virus infection. AB - Cross-sectional studies reported that hepatitis B core gene mutations are associated with active liver disease and responsiveness to interferon therapy. In view of the heterogeneity in published sequences, it is not possible to tell whether the differences in sequences observed were true mutations that developed during the course of infection. We conducted a longitudinal study to determine the rate of hepatitis B core gene mutations and the timing of these mutations in relation to hepatitis B virus replication, activity of liver disease, hepatitis B virus replication, activity of liver disease, hepatitis B e antigen (HBeAg) seroconversion, and interferon therapy. Serial sera from 55 patients with chronic hepatitis B infection were analyzed by direct sequencing of the hepatitis B precore/core gene to identify the nucleotide and amino acid changes that emerged during during follow-up. Patients who remained HBeAg positive and had normal amino transferase levels (Group I) maintained higher serum hepatitis B virus (HBV) DNA levels but significantly lower rates of both nucleotide and amino acid changes during follow-up compared with patients who remained HBeAg positive but had elevated aminotransferase levels (Groups II) and patients who cleard HBeAg (Group III). The rates of nucleotide and amino acid changes in Groups I, II, III patients were: 0.4 +/- 0.1, 1.9 +/- 0.3, and 2.4 +/- 0.4/nucleotide position/year; and 0.04 +/- 0.02, 0.21 +/- 0.05, and 0.38 +/- 0.07/codon/year, respectively. Most of the amino acid changes in Groups II and III patients occured during or shortly after flares in amino transferase levels, before HBeAg seroconversion. Similar rates of nucleotide and amino acid changes were found in interferon treated versus untreated patients, and in responders versus nonresponders. There was no difference in the location or nature of the hepatitis B core gene mutations between patients with and without HBeAg seroconversion, interferon treated versus untreated patients, and interferon responders versus nonresponders. In summary, changes in the hepatitis B core gene sequence was rarely detected in patients who were still in the immune tolerant phase but very high rates of changes were found during the immune clearance of chronic hepatitis B infection. Interferon therapy did not induce a higher rate or specific pattern of mutations in the hepatitis B core gene. Response to interferon therapy in HBeAg positive patients was unrelated to the number or location of hepatitis B core gene mutations. PMID- 8707279 TI - Elevated aflatoxin exposure and increased risk of hepatocellular carcinoma. AB - To elucidate the importance of aflatoxin in the etiology of hepatocellular carcinoma (HCC), a community-based cohort study combined with molecular dosimetry of aflatoxin exposure was performed in the Penghu Islets where the HCC mortality rate is highest in Taiwan. A total of 6,487 residents aged 30 to 65 years were recruited in the two-stage screening survey and underwent regular follow-up examination. Among 33 newly diagnosed HCC cases, 31 (94%) were chronic hepatitis B surface antigen (HBsAg) carriers and 3 (9%) were positive for antibodies against hepatitis C virus (HCV). Among 20 HCC patients and 86 matched healthy controls whose serum samples were tested for aflatoxin B1 (AFB1)-albumin adducts by competitive enzyme-linked immunosorbant assay (ELISA), 13 (65%) HCC patients and 32 (37%) matched controls were seropositive, showing a statistically significant multivariate-adjusted odds ratio of 5.5 with a 95% confidence interval of 1.2 to 24.5. The results imply the elevated risk of HCC among Penghu residents may be attributable to their heavy exposure to aflatoxins and high HBsAg carrier rate. PMID- 8707280 TI - The ancestral hemochromatosis haplotype is associated with a severe phenotype expression in Italian patients. AB - We evaluate the relation between genotype and phenotype in 47 Italian male patients with homozygous genetic hemochromatosis (GH). Phenotype evaluation was based on the ratio of amount of iron removed (IR) by phlebotomy and age (IR/age). Patients were divided in two classes of phenotype expression: class I included 26 patients with less severe iron overload (IR/age <0.33) and class II included 21 patients with a more marked one (IR/age >0.33). Genetic variability was assessed by haplotype analysis combining alleles at HLA-B, D6S265, HLA-A, and D6S105 loci. A common ancestral haplotype carrying D6S265-1, HLA-A3, and D6S105-8 alleles was present in 13 of 52 (25%) chromosomes in class I and in 24 of 42 (57%) chromosomes in class II (P = .0027). Homozygotes and heterozygotes for the ancestral haplotype had higher iron indices than patients carrying two haplotypes other than the ancestral one. Seven of the eight patients homozygous for the ancestral haplotype were in class II, heterozygotes were equally distributed between the two classes, whereas 14 of 18 carriers of other haplotype combinations were in class I. Our results suggests that the gene defect linked to the ancestral haplotype is the result of a single, severe mutation. The high variability of phenotype expression in heterozygotes for the ancestral haplotype could be accounted for the contribution of the mutation carried by the second haplotype. Combination of different mutations could be responsible for the variable degrees of iron overload found in patients with GH. PMID- 8707281 TI - Systemic hemodynamics and serum nitrate levels in patients undergoing endoscopic variceal ligation. AB - The effects of endoscopic variceal ligation (EVL) on systemic hemodynamics are unknown. This study was conducted to determine whether the obliteration of portal systemic collaterals by EVL affects systemic hemodynamics and serum nitrate concentrations in patients with compensated cirrhosis. We measured systemic and hepatic hemodynamics, azygos vein blood flow (AzBF), and serum nitrate concentrations before and immediately following EVL. A prompt decrease in left ventricular end-diastolic volume (LVEDV), stroke volume, cardiac index (CI), and an increase in systemic vascular resistance index (SVRI) were observed following variceal ligation. The reduction in LVEDV was associated with a decline in CI with a rise in SVRI. There was also a prompt reduction in serum nitrate concentration following variceal ligation. AzBF also significantly decreased following variceal ligation. These findings indicate that EVL decreased cardiac output via a reduction in cardiac preload (central venous return). Blood flow through portal-systemic collaterals has an important role in the enhanced cardiac preload of cirrhotic patients. The sudden decrease in serum nitrate concentrations suggests that endogenous nitric oxide may be involved in the regulation of systemic hemodynamics in patients with compensated cirrhosis. PMID- 8707282 TI - Albumin but not fibrinogen synthesis correlates with galactose elimination capacity in patients with cirrhosis of the liver. AB - Albumin and fibrinogen synthesis rates were measured in 15 subjects with different clinical stages of postviral cirrhosis and compared with galactose elimination capacity and aminopyrin breath test. Forty-three mg per kg body weight [2H5ring]phenylalanine with an isotopic enrichment of 10 atom% were intravenously injected. [2H5ring]phenylalanine enrichments in the plasma-free phenylalanine and the albumin and fibrinogen isolates were measured by gas chromatography-mass spectrometry. Fractional synthesis rates of albumin were normal in Child A cirrhosis (7.6 +/- 2.2%d), but were lower in both Child B (3.5 +/- 0.8%d) and C (4.5 +/- 2.8%d). Absolute rates of albumin synthesis were (103 +/- 30 mg/kg/d) in the child A group and substantially lower in the Child B (50 +/- 3 mg/kg/d) and C (36 +/- 20 mg/kg/d) group. The average fractional synthesis rate of fibrinogen was 16.7 +/- 7.5%d and the absolute synthesis rate 11.6 +/- 6.4 mg/kg/d. The values of the galactose elimination capacity and the aminopyrin breath test were below the normal range in all patients, gradually decreasing with an increase in the severity of the clinical stage of cirrhosis. Albumin synthesis rates significantly correlated with the Child scores, the galactose elimination capacity, and the aminopyrin breath test, whereas fibrinogen synthesis rates showed no such correlations. PMID- 8707283 TI - Immunoregulatory cytokines in chronic hepatitis C virus infection: pre- and posttreatment with interferon alfa. AB - T lymphocytes and immunoregulatory cytokines may be important in the host response to hepatitis C virus (HCV) infection. T-helper type 1 (Th1) cytokines (interleukin [IL]-2, interferon gamma [IFN-gamma]) are required for host antiviral immune responses, including cytotoxic T-cell generation and natural killer cell activation, while T-helper type 2 (Th2) cytokines (IL-4,IL-10) can inhibit the development of these effector mechanisms. In this study, the serum levels of Th1 and Th2 cytokines in patients (n = 23) infected with HCV were measured and compared with biochemical (alanine transaminase [ALT]) and viral (HCV RNA) indicators of infection. Serial cytokine levels were measured in a subset of 11 patients at 1 and 12 weeks during and at 1 week after interferon alfa (IFN-alpha) therapy (n = 33 samples). Levels of circulating IL-2, IL-4, IL 10, and IFN-gamma were significantly elevated in HCV patients versus normal controls (128 vs. 25 pg/mL, 3,045 vs. 29 pg/mL, 2,949 vs. 18 pg/mL, and 307 vs. 24 pg/mL respectively; P < .01). Treatment with IFN-alpha decreased the levels of IL-4 (321 +/- 224 pg/mL), and IL-10 (1,011 +/- 344 pg/mL), which paralleled a decrease in HCV RNA (114 +/- 27 vs. 25 +/- 20 Eq/ml X 10(5), pre- vs. post-IFN alpha [12 weeks];P <.05). These findings indicate that an activated T-cell response, as manifest by increased circulating immunoregulatory cytokines, is present in patients with HCV liver disease. Furthermore, treatment with HCV liver disease. Furthermore, treatment with IFN-alpha diminishes the Th2 cytokine response. Thus, modulation of T-cell function and cytokine production may be one mechanism whereby IFN-alpha therapy results in reduced viral burden. PMID- 8707284 TI - Liver hepatocyte growth factor does not always correlate with hepatocellular proliferation in human liver lesions: its specific receptor c-met does. AB - Increased levels of expression of hepatocyte growth factor (HGF) and its specific receptor c-met have been shown in the liver of several benign and malignant pathologies, both in experimental models and humans. We investigated by immunohistochemistry the presence of both HGF and c-met protoocogene product (c met pp) in 20 hepatocellular carcinomas (HCCs), 5 focal nodular hyperplasias (FNHs), 4 cases of fulminant hepatitis (FH), and 1 case of regenerated liver. The c-met protooncogene product was expressed in all cases with marked overexpression in the HCCs and in ductular metaplasia. HGF was detected in the Ito cells of all cases and in neoplastic hepatocytes of 9 of 20 HCCs (45%). The proliferative index of each lesion was evaluated by means of the polyclonal antibody anti cyclin A. When the level of expression of HGF and c-met protooncogene product with the percentage of cyclin A+ nuclei were compared, the closest relationship was between c-met protooncogene product and cyclin A+ nuclei were compared, the closest relationship was between c-met protooncogene product and cyclin A. In 11 of 20 HCCs (55%), there wa no correlation between HGF positivity and cyclin A. This seems to suggest that, independently of the levels of native liver HGF, c met protooncogene product is the most active modulator of liver cell proliferation. PMID- 8707285 TI - Variable gene expression within human tyrosinemia type 1 liver may reflect region specific dysplasia. AB - Patients with hereditary tyrosinemia type 1 have a deficiency of fumarylacetoacetate hydrolase (FAH) and develop progressive hepatocellular dysfunction with a high risk of malignant transformation. Serum alpha-fetoprotein levels are frequently elevated in these patients; therefore, this commonly used marker of tumorigenesis is inadequate. To date, no literature exists describing the hepatic gene alterations in patients with this disease. We analyzed the expression of a panel of proliferation associated and liver-specific genes in the liver of a 33 month-old girl at the time of orthotopic liver transplantation. This study provides information that may be useful in developing markers for malignancy and understanding the pathogenesis of this disease. Gene expression patterns of two regenerating nodules and total liver from the patient with FAH deficiency were compared with control donor liver. Liver-specific and growth induced genes with altered expression in the tyrosinemic liver included several functional classes: structural proteins (actin, thrombospondin), transcription factors (c-fos, egr-1, C/EBPalpha), liver-specific enzymes (glucose-6-phosphatase [G6Phase], and secreted factors (insulin-like growth factor binding protein 1 [GFBP-1]. Isolated macronodules demonstrated varied patterns of expression, suggesting that they do not form a homogeneous cellular environment. In the tyrosinemic liver, IGFBP-1 messenger RNA expression was high and G6Phase messenger RNA was not detectable. Although G6Phase and IGFBP-1 are coexpressed in regenerating liver, immunohistochemistry in the tyrosinemic liver demonstrated a mutually exclusive distribution for the two proteins in a tissue section with features of dysplasia. We propose that cells in these areas may have an aberrant transcription factor and growth factor "milieu" that leads to altered gene and protein expression. These molecular alterations are reflected in dysplastic histologic changes and may ultimately predispose to the development of malignancy. PMID- 8707286 TI - Enhanced expression of the protein kinase substrate annexin in human hepatocellular carcinoma. AB - Annexin (AX) constitutes a new family of Ca2+-dependent membrane-binding proteins; 13 of them have been described. Among these, annexin-1 (AX-I) has displayed many biological functions in vitro. Its actual role in vivo, however, remains unknown. We already reported that AX-I was expressed in proliferating (regenerating) hepatocytes at both protein and messenger RNA (mRNA) levels. The role of AX-I in human hepatocellular carcinoma (HCC) remains obscure. In this study, the amounts of AX-I at protein and mRNA levels, as well as its localization, have been determined in the normal human liver, chronic hepatitis liver, and nontumorous and tumorous portions of HCC. AX-I was rarely found in normal and chronic liver tissues, whereas it is overexpressed at both the transcriptional and translational levels in tumorous and nontumorous regions of HCC. In addition, more AX-I was expressed in the tumorous portion than the nontumorous portion of HCC. AX-I was present in the hepatocytes and HCC cells, localized mainly in the cytoplasm. AX-I was expressed in poorly differentiated cancer cells. Furthermore, AX-I was tyrosine-phosphorylated in HCC. We also found that some of the AX-I- positive hepatocytes in the nontumorous tissues were derived from a particular subset of parenchymal cells (stem or oval cells). These results indicate that AX-I plays an important role in the malignant transformation process leading to HCC and that it is closely related to the histological grade of HCC. HCC would offer a novel tool with which to study the function of AX-I in malignant transformation. PMID- 8707287 TI - Cellular distribution of transcripts for tissue inhibitor of metalloproteinases 1 and 2 in human hepatocellular carcinomas. AB - The cellular distribution of tissue inhibitor of metalloproteinases (TIMP)-1, and TIMP-2 was studied by using in situ hybridization in surgically removed human hepatocellular carcinomas (HCCs) and cholangiocellular carcinomas (CCCs). The purpose of this study was to characterize the protein involvement of TIMPs in the development of HCCs and CCCs. All HCCs and CCCs expressed TIMPs. The distribution of transcripts for TIMPs in the tumors was mostly homogeneous. Expression of TIMP in cancer cells was more intense than that in the surrounding noncancerous liver (either, cirrhosis, chronic hepatitis, or normal), and expression of TIMP-1 was stronger than that of TIMP-2. Expression of TIMPs varied among HCC nodules, but there was no obvious association between the expression level of TIMPs and differentiation stages or invasiveness of the HCCs. Transcripts for TIMPs were clearly demonstrated in the metastatic HCC nodules in the lung. Expression of TIMP-1 CCC was strong, and small nodules of CCC were recognized in the liver. Immunohistochemical study for TIMP-1 revealed a consistent staining of the TIMP protein with the transcripts. In the peritumoral histologically normal liver, which was not infected with either hepatitis B or C virus, expression of TIMP-1 was found in various cell types, but that of TIMP-2 was weak. Expression of TIMP 1 in hepatocytes revealed clear zonal distribution. These results suggest that TIMPs may act on modulating the matrix/tumor interaction and may play an important role in growth and invasion of HCCs and CCCs. Expression of TIMP-1 can be a marker of HCC metastasis to the lung, and also that of the extent of CCC invasion. Furthermore, the consistent expression of TIMPs in many cell types of the noncancerous liver suggests some unknown functional role that must be clarified. PMID- 8707288 TI - Donor age and outcome of liver transplantation. AB - To evaluate the effect of donor age on graft and patient outcome after liver transplantation an analysis of a large-scale cohort study was performed at three tertiary referral liver transplant centers. Between April 1990 and June 1994, 772 adults underwent an initial single-organ liver transplantation. The age of the donors averaged 35 years;193 (25%) were 50 or above, the age used to define "older" donors. Groups were compared for demographic, clinical, and biochemical features. Outcome was measured using results of biochemical tests after transplantation and by graft and patient survival. Compared with younger donors, older donors were more commonly women (59% vs. 33%:P < or = .001) and died of central nervous system causes (79% vs. 28%) as opposed to trauma (13% vs. 63%:P < or = .001). The recipients of the two groups of donor livers did not differ in important respects. However, intraoperatively, livers from older donors were more likely to be assessed as either "poor" or "fair" as opposed to "good" (17% vs. 4%:P < or = .001) by the harvesting surgeon and to have initial "poor" or "fair" bile production (29% vs. 18%:P < or = .001). During the first week postoperatively, the serum aminotransferase and bilirubin levels and prothrombin times were higher in recipients of older than those of younger donor livers. During follow-up, graft survival was less for recipients of older donor livers at 3 months (81% vs.91%:P = .0001) and at 1 (76% vs. 85%:P = .007) and 2 years (71% vs. 80%:P = .005); patient survival showed similar though less marked differences. This association of donor age and poorer graft survival persisted after adjusting for many variables using bivariate and multivariate analyses. Importantly, however, the association with poor graft survival was largely among recipients of older donor livers, the quality of which was assessed as fair or poor by the harvesting surgeon; recipients of older donor livers assessed as good had a retransplant-free survival similar to that of younger donor livers (87% vs. 91% at 3 months). Thus, use of older donor livers, the quality of which are judged to be good by the harvesting surgeon, is not associated with a decrease in patient or graft survival after liver transplantation. differences. This association of donor age and poorer graft survival persisted after adjusting for many variables using bivariate and multivariate analyses. Importantly, however, the association with poorer graft survival was largely among recipients of older donor livers, the quality of which was assessed as fair or poor by the harvesting surgeon; recipients of older donor livers, the quality of which was assessed as fair or poor by the harvesting surgeon; recipients of older donor livers assessed as good had a retransplant-free survival similar to that of younger donor livers (87% vs. 91% at 3 months). Thus, use of older donor livers, the quality of which are judged to be good by the harvesting surgeon, is not associated with a decrease in patient or graft survival after liver transplantation. PMID- 8707289 TI - Use of a designer triple expression hybrid clone for three different lipoyl domain for the detection of antimitochondrial autoantibodies. AB - The detection of antimitochondrial autoantibodies (AMAs) is critical in the diagnosis of primary biliary cirrhosis (PBC). However, conventional laboratory assays to detect AMA are dependent on the time-consuming method of immunofluorescence microscopy, a method often plagued by problems of nonspecificity. AMAs react against mitochondrial autoantigens including the E2 components of the pyruvate dehydrogenase complex (PDC-E2), the branched-chain 2 oxo-acid dehydrogenase complex (BCOADC-E2), and the 2-oxo-glutarate dehydrogenase complex (OGDC-E2). Interestingly, the immunodominant epitopes of PDC-E2, BCOADC E2, and OGDC-E2 are all conformational lipoate binding sites, but antibodies against them do not cross-react. Although 80% to 90% of sera from patients with PBC react to PDC-E2, approximately 10% patients with PBC react only to BCOADC-E2 and/or OGDC-E2. We have taken advantage of our epitope-mapping studies of the E2 components of PDC, BCOADC, and OGDC, and constructed a "designer" hybrid clone, designated as pML-MIT3, that coexpresses the immunodominant epitopes within the three distinct lipoyl domains. We examined a total of 321 sera, including 186 sera from patients with PBC, to test the immunoreactivity of pMIT3. Of 186 sera from patients with PBC, 152 sera (81.7%) reacted with recombinant fusion protein of PDC-E2, whereas 171 sera (91.9%) showed positive reactivities when probed by immunoblotting against the recombinant fusion protein expressed from the pML-MIT3 clone. Of 34 PBC sera that did not react with recombinant PDC-E2, 18 sera contained BCOADC-E2-specific AMA and 1 serum possessed only OGDC-E2-specific AMA. We also developed an enzyme-linked immunosorbent assay (ELISA), using affinity purified recombinant fusion protein of pML-MIT3 clone as the antigen source, to quantify specific AMAs in patients with PBC. None of the 135 control sera from patients with primary sclerosing cholangitis (PSC), chronic autoimmune hepatitis (CAH), systemic lupus erythematosus (SLE), or healthy volunteers showed significant reactivity against pML-MIT3 recombinant fusion protein in the ELISA assay. Our results indicate that an ELISA using recombinant, cloned autoantigen of pML-MIT3 is a powerful and very specific method for the detection of AMA. PMID- 8707290 TI - Detection of amplified genomic sequences in human small-cell lung carcinoma cells by arbitrarily primed-PCR genomic fingerprinting. AB - The arbitrarily primed-PCR (AP-PCR) genomic fingerprinting method was applied to evaluate its effectiveness in detecting and characterizing amplified DNA fragments in two small-cell lung carcinoma (SCLC) cell lines, NCI-H69 and NCI H82. Of the 2428 DNA fragments detected by AP-PCR using 62 arbitrary primers, 2 (0.08%) DNA fragments were amplified in NCI-H69 and 6 (0.25%) DNA fragments were amplified in NCI-H82. Based on these results, we estimate the total size of the amplified genomic regions in these cell lines to be 3000 megabase pairs (Mb) x 0.0008 = 2.4 Mb in NCI-H69 and 3000 Mb x 0.0025 = 7.5 Mb in NCI-H82. The 2 amplified fragments in NCI-H69 were mapped to chromosome 2, and all 6 amplified fragments in NCI-H82 were mapped to chromosome 8. This strongly suggests that restricted chromosomal regions are specifically amplified in these SCLC cell lines. Since the N-myc gene at 2p24 is amplified in NCI-H69 and the c-myc gene at 8q24 is amplified in NCI-H82, it is possible that these DNA fragments are co amplified with N-myc or c-myc in these cell lines. However, since the 2 amplified fragments in NCI-H69 were not amplified in 42 other human cancer cell lines including 11 cell lines carrying amplified N-myc genes, it is also possible that there are amplified regions on chromosome 2 other than the N-myc locus at 2p24 in NCI-H69. In contrast, all 6 amplified fragments in NCI-H82 were amplified in several other human cancer cell lines carrying amplified c-myc genes. This result further indicates that these fragments were derived from an amplification unit that includes the c-myc gene. Our results show the ability of the AP-PCR method to analyze the fraction of the genome with amplification in human cancer cells. PMID- 8707291 TI - Genetic basis of Creutzfeldt-Jakob disease in the United Kingdom: a systematic analysis of predisposing mutations and allelic variation in the PRNP gene. AB - Creutzfeldt-Jakob disease (CJD) is a transmissible neurodegenerative disorder characterized by the accumulation of aggregates of a cellular protein, PrP, in the brain. In both human and animals, genetic alterations to the gene encoding PrP (PRNP in human) modulate susceptibility to CJD. The recent epidemic of bovine spongi-form encephalopathy in the UK has raised the possibility of transmission from animal produce to humans. To provide a baseline against which to assess possible risk factors, we have determined the frequencies of predisposing mutations and allelic variants in PRNP and their relative contributions to disease. Systematic PRNP genotype analysis was performed on suspected CJD cases referred to the National Surveillance Unit in the UK over the period 1990-1993. Inspection of 120 candidate cases revealed 67 patients with definite and probable CJD, based on clinical and neuropathological criteria. No PRNP mutations were detected in any of the remaining 53 patients assessed as "non-CJD". A disease associated mutation in the PRNP gene was identified in nine (13.4%) definite and probable cases of CJD, a reliable estimate of the incidence of PRNP-related inherited CJD based on a prospective epidemiological series. Within the group of sporadic CJD patients (lacking PRNP mutations), we confirmed that the genotype distribution with respect to the common methionine/valine (Met/Val) polymorphism at codon 129 within PRNP was significantly different from the normal Caucasian population. The incidence of Met homozygosity at this site was more than doubled and correlated with increased susceptibility to the development of sporadic CJD. Unlike other recent studies, Val homozygosity was also confirmed to be a significant risk factor in sporadic CJD, with the relative risks for the three genotypes Met/Met: Val/Val:Met/Val being 11:4:1. PMID- 8707292 TI - Characterization of double minute chromosomes' DNA content in a human high grade astrocytoma cell line by using comparative genomic hybridization and fluorescence in situ hybridization. AB - The presence of double minute chromosomes (dmin) in cancer cells is known to be correlated with gene amplifications. In human high grade astrocytomas or glioblastomas, about 50% of cytogenetically characterized cases display dmin. G5 is a cell line which has been established from a human glioblastoma containing multiple dmin. In order to identify the DNA content of these dmin, three techniques were successively used: conventional cytogenetic analysis, comparative genomic hybridization (CGH). and fluorescent in situ hybridization (FISH). The karyotype of G5 cells showed numerical chromosome changes (hypertriploidy), several marker chromosomes, and multiple dmin. CGH experiments detected two strong DNA amplification areas located in 9p21-22 and 9p24, as well as an underrepresentation of chromosomes 6, 10, 11, 13, 14, and 18q. By using FISH with a chromosome 9-specific painting probe to metaphase chromosomes of the G5 cell line, dmin were shown to contain DNA sequences originating from chromosome 9. This study demonstrates the usefulness of a combination of classical karyotyping, CGH, and FISH to identify the chromosomal origin of amplified DNA sequences in dmin. PMID- 8707293 TI - Mutations in the VHL tumor suppressor gene and associated lesions in families with von Hippel-Lindau disease from central Europe. AB - von Hippel-Lindau (VHL) disease is a dominantly inherited familial cancer syndrome predisposing to retinal, cerebellar and spinal hemangioblastoma, renal cell carcinoma (RCC), pheochromocytoma and pancreatic tumors. Clinically two types of the disease can be distinguished: VHL type 1 (without pheochromocytoma) and VHL type 2 (with pheochromocytoma). We report VHL germline mutations and trends in phenotypic variation in families from central Europe. We identified 28 mutations in 53/65 (81.5%) families with 18 (64%) mutations being unique to this population. Whereas types and distribution of mutations as well as a strong correlation of missense mutations with the VHL 2 phenotype were similar to those identified in other populations, these families have provided new insights into the molecular basis for variability in the VHL 2 phenotype. Seven different missense mutations in exons 1 and 3 varied in their biological consequences from a minimal VHL 2 phenotype with pheochromocytoma only to a full VHL 2 phenotype with RCC and pancreatic lesion. These findings contribute to a better understanding of the fundamental mechanisms of VHL disease and its phenotypic variability. Further, we have provided rapid VHL screening for the families in central Europe, which has resulted in improved diagnosis and clinical management. PMID- 8707294 TI - beta-Glucuronidase P408S, P415L mutations: evidence that both mutations combine to produce an MPS VII allele in certain Mexican patients. AB - Mucopolysaccharidosis type VII (MPS VII, Sly syndrome) is an autosomal recessively inherited lysosomal storage disease caused by a deficiency in beta glucuronidase. We identified and studied a novel allele containing two C-to-T transitions resulting in P408S and P415L alterations, which is present in homozygous state in one Mexican patient and in heterozygous state in another. None of the previous reports describing mutations in the MPS VII gene include Mexican patients. Expression of either of the mutations individually showed only modest effects on the properties of the enzyme. However, expression of the doubly mutant allele resulted in markedly reduced activity and rapid degradation in an early biosynthetic compartment. PMID- 8707295 TI - Polymorphisms of apolipoproteins A-IV and E in a Turkish population living in Germany. AB - Human apolipoproteins (apo) E and apo A-IV are polymorphic with significantly different allele frequencies among different ethnic groups. Whereas the variation at the apo E gene locus affects plasma cholesterol levels in all populations studied so far and is associated with longevity in Caucasians, the influence of the common apo A-IV polymorphism on plasma lipoproteins has not been unanimously accepted. We have therefore determined the common apo E and apo A-IV polymorphisms by isoelectric focusing, calculated the respective allele frequencies and studied their effects on plasma lipoproteins in a random sample of 240 nonrelated Turkish subjects (141 males, 99 females) living in Germany and originating from central and eastern Anatolia. When compared with the German population and other Caucasians in Europe a prominence of the apo epsilon 3 allele frequency (0.885) was accompanied by a decrease in the frequencies of both the apo epsilon 2 allele (0.048) and the apo epsilon 4 allele (0.067). Thus, the Turkish population studied here clustered with populations mainly from southern Europe and Japan, which have low epsilon 2 and epsilon 4 allele frequencies. Also, the frequency of the A-IV-1 allele was higher (0.967) and that of the A-IV 2 allele lower (0.033) in the Turkish subjects studied than in other populations. At an average level of total cholesterol of 194.5 +/- 45 mg/dl, no significant influence of the A-IV alleles on plasma lipoproteins was seen. However, apo E and apo B differed significantly between apo E phenotypes, with high levels of apo E and low levels of cholesterol and apo B in carriers of the epsilon 2 allele, and vice versa for the epsilon 4 allele. The average cholesterol excess for the epsilon 2 allele was -7.95 mg/dl, for the epsilon 3 allele, -1.34, and for the epsilon 4 allele, + 14.15 mg/dl. Thus, despite the unusual frequency distribution of the apo E alleles, their effects on plasma lipoproteins are within the range reported for other populations in Europe. PMID- 8707296 TI - A jumping Robertsonian translocation: a molecular and cytogenetic study. AB - We report a patient with mosaicism for two different Robertsonian translocations, both involving chromosome 21. She carries an unbalanced cell line with an i(21q) and a balanced cell line with a rob(21q22q). She is phenotypically normal but has two children who inherited the i(21q) and have Down syndrome. We demonstrate that both abnormal chromosomes are dicentric and that the proband's 21/21 rearrangement is an isochromosome formed from a maternally derived chromosome 21. We propose a model in which the i(21q) is the progenitor rearrangement in the proband, which subsequently participated in a nonreciprocal rearrangement characteristic of a jumping translocation. In addition, we review other cases of constitutional mosaicism involving jumping translocations. PMID- 8707297 TI - A LINE element is present at the site of a 300-kb deletion starting in intron 10 of the PAX6 gene in a case of familial aniridia. AB - We have identified a 300-kb germline deletion in 11p13 in a family with aniridia but no Wilms' tumor. Cloning and sequencing of the breakpoint revealed that the deletion starts in intron 10 of the PAX6 gene and removes the C-terminal part of the proline-serine-threonine rich domain, leaving both DNA-binding domains intact. The PAX6 gene is joined head-to-head to a LINE-1 (L1) element. The L1 is truncated at the 3' end, removing part of ORF2. Sequencing of the L1 element shows that it does not encode a functional transposase and is therefore probably not an active element. These data suggest that the L1 element is normally present at the site of the distal deletion endpoint in 11p13. No extensive sequence homologies are detected at the deletion junction points; however, the PAX6 gene as well as the L1 element have runs of T nucleotides at this position, indicating that the deletion occurred by nonhomologous recombination. Several consensus recognition sequences for topoisomerase I flank the deletion site in both sequences, suggesting an involvement of this enzyme during the deletion recombination process. PMID- 8707298 TI - Haplotype identity between individuals who share a CFTR mutation allele "identical by descent": demonstration of the usefulness of the haplotype-sharing concept for gene mapping in real populations. AB - Cystic fibrosis (CF) patients with the A455E mutation, in both the French Canadian and the Dutch population, share a common haplotype over distances of up to 25 cM. French Canadian patients with the 621+1G-->T mutation share a common haplotype of more than 14 cM. In contrast, haplotypes containing the delta F508 mutation show haplotype identity over a much shorter genomic distance within and between populations, probably because of the multiple introduction of this most common mutation. Haplotype analysis for specific mutations in CF or in other recessive diseases can be used as a model for studying the occurrence of genetic drift conditional on gene frequencies. Moreover, from our results, it can be inferred that analysis of shared haplotypes is a suitable method for genetic mapping in general. PMID- 8707299 TI - Heteroduplex and protein truncation analysis of the BRCA1 185delAG mutation. AB - We describe a heteroduplex analysis for the detection of the 185delAG mutation in the BRCA1 gene. The protein truncation test (PTT) has previously been used to identify many of the mutations in BRCA1 that result in premature termination of the protein. However, we were not able to detect the 185delAG mutation by PTT and suggest that heteroduplex analysis may complement PTT for analysis of BRCA1 mutations. This simple technique may be useful for studies on the prevalence and the penetrance of the 185delAG mutation. PMID- 8707300 TI - Identification of a common 6-pyruvoyl-tetrahydropterin synthase mutation at codon 87 in Chinese phenylketonuria caused by tetrahydrobiopterin synthesis deficiency. AB - Deficiency in 6-pyruvoyl-tetrahydropterin synthase (PTPS) activity is the major cause of tetrahydrobiopterin (BH4)-deficient phenylketonuria. Two single base alterations of PTPS cDNA, a C-to-T transition at nucleotide 259 and a novel A-to G transition at nucleotide 155 (according to cDNA sequence), were identified in two Chinese PTPS-deficient siblings by the reverse transcription-polymerase chain reaction (RT-PCR). The C-to-T transition at nucleotide 259 results in an amino acid change from proline to serine at codon 87 (Pro87Ser), and the A-to-G transition at nucleotide 155 causes an amino acid change from asparagine to serine at codon 52 (Asn52Ser) of PTPS. The C259T missense mutation can be identified by analysis of the BbvI restriction fragments of the PCR-amplified PTPS cDNA product, and was found to account for 42% (11/26) of 26 Chinese PTPS mutant alleles studied. However, none of 100 normal alleles screened were found to have this change. This result indicates that the C259T transition may be a common mutation in Chinese PTPS-deficient patients. PMID- 8707301 TI - Anesthesiologic problems in Williams syndrome: the CACNL2A locus is not involved. AB - We present the case of a patient affected with Williams syndrome (WS), who developed a suspected malignant hyperthermia (MH) reaction to general anesthesia. The proximity to the WS region of the gene encoding the L-type voltage-gated calcium channel alpha 2/delta-subunit (CACNL2A) on 7q11.23-q21.1, previously shown to be closely linked to some forms of MH susceptibility, prompted us to investigate whether this gene is deleted in WS. Linkage studies and fluorescence in situ hybridization analysis demonstrated that the CACNL2A locus is localized outside the WS deleted region. PMID- 8707302 TI - Yeast artificial chromosome mapping of the cystinosis locus on chromosome 17p by fluorescence in situ hybridization. AB - The gene locus for cystinosis has been mapped between markers D17S1583 and D17S1584 on the short arm of chromosome 17. Using markers encompassing the cystinosis region, we assigned different yeast artificial chromosome (YAC) clones previously identified by sequence tagged site (STS) screening to 17p13.3. Three of the clones hybridized to the target 17p gene region; one of these was chimeric, hybridizing both to chromosomes 3p and 5q; two of the YACs did not contain sequences of 17p13.3. Our physical mapping has identified candidate YACs as a first step towards a positional cloning approach. PMID- 8707303 TI - Fragile site and interstitial telomere repeat sequences at the fusion point of a de novo (Y;13) translocation. AB - We describe a novel fragile site in a rearranged chromosome, associated with the presence of telomeric repeat sequences at the fusion point of a translocation between chromosomes 13 and Y. The case reported in this study shows a de novo (Y;13) translocation, which appears to represent fusion of an apparently intact chromosome Y with a chromosome 13 that has lost only part of its short arm. Ten percent of the cells show a normal karyotype without the (Y;13) translocation. Molecular cytogenetic studies of the derived Y;13 chromosome revealed three hybridization sites of the telomeric probes-one at each end and one at the breakpoint junction. A fragile site is also observed in the intrachromosomic telomeric region. This coincidence suggests that the telomere repeat sequences (TTAGGG)n, when present at an interstitial chromosomal location, can promote the formation of a novel fragile site. PMID- 8707304 TI - Transcript analysis of CFTR frameshift mutations in lymphocytes using the reverse transcription-polymerase chain reaction technique and the protein truncation test. AB - mRNA transcripts of the cystic fibrosis transmembrane conductance regulator (CFTR) gene were analyzed from lymphocytes of two cystic fibrosis compound heterozygotes (394delTT/3195del6 and 1215delG/ 2423delG), of five related carriers heterozygous for one of these mutations, and of five normal individuals. After reverse transcription of total RNA and amplification by the polymerase chain reaction, fragments were investigated by sequencing and by the protein truncation test (PTT). The three frameshift mutants were correctly detected by PTT, as they introduced a premature termination codon resulting in shortened protein products. The PTT approach thus provides a simple and reliable alternative method for detecting frameshift, nonsense, or splice site mutations, and for ascertaining their putative effect on the reading frame of the mRNA. In addition, we have identified 6 alternatively spliced forms of CFTR mRNA, two of which (transcripts lacking 4 + 5 or 17B) have not been described previously. PMID- 8707305 TI - Identification of a variable number tandem repeat region in the human T cell receptor alpha-delta (TCRAD) locus. AB - A number of different polymorphisms have been observed in coding as well as in non-coding regions of T cell receptor (TCR) genes. We report the identification and characterization of a highly polymorphic locus in the 3' noncoding region of the human T cell receptor alpha/delta (TCRAD) on chromosome 14. In 202 unrelated individuals, ten different alleles were distinguished by polymerase chain reaction (PCR) and a heterozygosity rate of 64% was calculated. Sequence analysis revealed that this polymorphic region consists of 10 bp imperfect repeat units and represents a variable number tandem repeat region (VNTR). Stable Mendelian inheritance of this novel polymorphic marker was proven in four families. The localization of this VNTR polymorphism in the TCRAD locus should make it a useful system for linkage analysis in immunological disorders with a known role of TCRAD. PMID- 8707306 TI - CFTR haplotypic variability for normal and mutant genes in cystic fibrosis families from southern France. AB - In order to contribute to a better understanding of the dispersion of cystic fibrosis (CF) mutations in the South of France, seven diallelic and three multiallelic markers [three upstream of the cystic fibrosis transmembrane conductance regulator (CFTR) gene (XV-2c, KM 19 and J44) and seven intragenic polymorphism (IVS6A, IVS8CA, M470V, T854T, IVS17BTA, IVS17BCA and TUB18)] were analyzed for 143 delta F508 chromosomes, 100 CF chromosomes carrying 85 non-delta F508 and 15 unknown mutations, and 198 normal CFTR alleles. The study provides haplotypic data for 39 different CF mutations, which should be useful in diagnosis by haplotypic analysis and detection of the associated mutations. A major haplotype [2-1-2-7-16-2-1-(30/31)-13-1] was found in normal chromosomes, which should be the most ancient in the Caucasoid population. The most frequent haplotypes in normal chromosomes were associated with 16 different non-delta F508 mutations, suggesting that there was no preferential haplotype on which these mutations arose. Several mutations were each associated with more than one haplotype, as the result of slippage at one or two of the three microsatellites (delta F508, G542X, N1303K, G85E, E585X, K710X and 2184delA) or recombination (1717-1G-->A, R334W, L206W, R1162X and Y122X). Haplotypes for the most common CFTR mutations (delta F508, G542X, N1303K) revealed that a large number of alleles were generated by slippage at the microsatellite loci, suggesting that they are the most ancient CF mutations. Other mutations were associated with haplotypes that were different either at several diallelic sites (R334W) or at both diallelic and microsatellite markers (R1162X and R1158X), which is more suggestive of recurrence. Twenty recombinations were detected among the CF mutant alleles analyzed, 75% of them occurring in the second half of the CFTR gene. The higher mutational heterogeneity and the haplotypic variability reported in this small population from the Mediterranean area are consistent with an earlier appearance of CFTR mutations in southern Europe than in central and northern Europe, and an earlier origin and expansion of this population. PMID- 8707307 TI - Prevalence of alpha-thalassemias in northern Thailand. AB - The population of northern Thailand has one of the highest frequencies of alpha thalassemia in the world. However, the available distributional data are controversial. In addition to deletional types of alpha-thalassemia Hb, type Constant Spring should also be taken into consideration in alpha-thalassemia population studies, because it causes clinical alpha-thalassemia in the homozygous state or when present with both alpha-globin genes deleted in trans. We have examined a sample of 215 healthy subjects from four rural districts of Chiang Mai province. Out of these, 77 exhibited anomalies of the alpha-globin genes (alpha alpha/-alpha 3.7 in 36; -alpha 3.7/-alpha 3.7 in 3; -SEA in 30; alpha alpha/alpha CS alpha in 5; alpha alpha alpha anti 3.7 in 3). Therefore, no fewer than 2% of the children in northern Thailand are expected to be born with HbH disease or thalassemic hydrops fetalis. The considerable public health problem of hemoglobinopaties and the increasing acceptance of family planning necessitates facilities for the pre- and postnatal diagnosis of these disorders at the DNA level. PMID- 8707308 TI - A new polymorphism of arylsulfatase A within the coding region. AB - A 10-year-old boy with juvenile metachromatic leukodystrophy (MLD) presented with the 459 + 1G-->A arylsulfatase A (ASA) mutation on one allele. To detect his complete genotype, the other ASA allele was sequenced and a T-to-C transition at nucleotide 376 in exon 2 was identified. This missense mutation results in a substitution of leucine 76 by proline. Of 20 MLD unrelated controls, 18 carried the L/P76 mutation either in the homozygous (n = 6) or heterozygous (n = 12) state. The presence or absence of L/P76 did not influence leukocyte ASA activity or urinary sulfatide excretion. Apparently, the substitution of leucine 76 by proline is a common ASA polymorphism, neither being related to MLD nor creating ASA pseudodeficiency. However, because of its frequency and location, L/P76 may be of particular importance in genetic studies requiring the differentiation of the ASA alleles within a kindred. Further studies are directed to the as yet unresolved genotype of the index case with juvenile MLD. PMID- 8707309 TI - Molecular reconstruction and homology modelling of the catalytic domain of the common ancestor of the haemostatic vitamin-K-dependent serine proteinases. AB - The vitamin-K-dependent serine proteinases of coagulation have evolved by a process of gene duplication and divergence, acquiring along the way a considerable degree of functional diversity that has equipped them for their different roles in haemostasis. The cDNA sequences encoding the catalytic domains of the early mammalian ancestors of five vitamin-K-dependent factors (factors VII, IX and X, protein C and prothrombin) were reconstructed by employing cDNA sequence data from a range of extant mammals and by using established phylogenies. The cDNA sequence of the putative common ancestor of these early mammalian proteins was then reconstructed from the five sequences by using a deduced phylogeny that was different in a number of respects from those previously proposed. Factor IX is proposed to have branched off early on, followed by protein C and prothrombin and finally factors VII and X. Significant differences in mutation rates were observed between proteins within a species; factor IX exhibited a lower mutation rate than the other proteins, consistent with its early emergence. Differences in mutation rates were also observed between species for a given protein and these exhibited an inverse correlation with generation time. A biophysically plausible structure for the ancestral vitamin-K-dependent factor protein was constructed by comparative methods. Studies of the functional architecture of this model provide new insights into the evolution of protein-binding specificity in this family of proteins. PMID- 8707310 TI - Autosomal dominant spastic paraplegia with anticipation maps to a 4-cM interval on chromosome 2p21-p24 in a large German family. AB - Autosomal dominant familial spastic paraplegias (AD-FSP) are a group of genetically heterogeneous diseases characterised by a progressive spasticity of the lower limbs. Three loci have already been identified by genetic linkage studies on chromosomes 2p, 14q and 15q. Here we present linkage data from a large German family displaying AD-FSP with anticipation which confirms the existence of the FSP2 locus on chromosome 2p. The recombination events observed in our family define the critical region for the FSP2 gene to be within a 4-cM interval, flanked by markers D2S400 and D2S367. Moreover, clinical data from our family show evidence of anticipation, a phenomenon caused by trinucleotide expansion in several other neurodegenerative diseases. PMID- 8707311 TI - Low frequency of deletion alleles in patients with steroid 21-hydroxylase deficiency in a Mexican population. AB - Steroid 21-hydroxylase deficiency is caused by mutations in the CYP21 gene. Approximately 95% of mutant alleles are generated by recombination events between the active gene CYP21 and its highly homologous pseudogene, CYP21P. Deletion alleles are generated by unequal crossing over, while point mutations are the result of gene conversion events. Deletions account for 20-25% of the 21 hydroxylase deficiency alleles in most populations studied. We have looked for deletions among 53 unrelated Mexican patients with steroid 21-hydroxylase deficiency and found that deletions represent less than 1% of the disease alleles. These findings suggest that nearly all mutant alleles in our patient population contain point mutations and that the low representation of deletion alleles among clinically diagnosed patients may be due to missing detection of salt wasters, mainly males, who may die during the neonatal period. PMID- 8707312 TI - Recessive Schwartz-Jampel syndrome (SJS): confirmation of linkage to chromosome 1p, evidence of genetic homogeneity and reduction of the SJS locus to a 3-cM interval. AB - Schwartz-Jampel syndrome (SJS), or chondrodystrophic myotonia, is a rare autosomal recessive disorder characterized by generalized myotonia resulting in a particular, recognizable facies and osteoarticular abnormalities. Some of us have recently shown genetic linkage of SJS to a locus on 1p34-p36.1 in five families. Here, we show by homozygosity mapping and segregation analysis that eight new families are most likely linked to the SJS locus on chromosome 1, confirming the localization of SJS to chromosome 1p and suggesting genetic homogeneity. Recombination events reduced the SJS locus from a genetic interval of 8 to 3 cM, which should facilitate the identification of the SJS gene. Low clinical variability was observed between the studied families, except for osteoarticular abnormalities. Since the severity and the location of osteoarticular abnormalities varied from one individual to another, even in the same families, other factors than the SJS gene itself, genetic or epigenetic, might contribute to the phenotype. PMID- 8707313 TI - Secretory phospholipase A2 does not appear to be associated with phenotypic variation in familial adenomatous polyposis. AB - Recent studies in mice have provided strong evidence for a modifier gene that is capable of effecting the expression of the mouse equivalent of familial adenomatous polyposis (FAP). A candidate gene has been proposed, namely secretory phospholipase A2 (sPLA2). Increased tumor number in mice was correlated with low levels of sPLA2 expression and the presence of truncating mutations within the sPLA2 gene. In an attempt to determine whether any genetic alterations in the sPLA2 gene were associated with the expression of FAP in man, we investigated the genetic structure of sPLA2 in 97 polyposis coli patients presenting with various disease phenotypes, and its expression in 8 FAP patients displaying markedly different disease characteristics. In the current study no inactivating mutations in the sPLA2 gene were identified, suggesting that human sPLA2 is not associated with phenotypic variation in FAP. PMID- 8707314 TI - Human papillomaviruses and cervical screening. PMID- 8707315 TI - Balanitis and balanoposthitis: a review. AB - OBJECTIVES: To give an overview of the literature on balanitis, with a special emphasis on infective causes. METHOD: A data search was performed using the OVID CD plus Medline 1967-1995, using balanitis and balanoposthitis as textword search strategy. Specific subjects such as anaerobic infection, Zoon's balanitis were sought separately and subgroups combined. Original articles and abstracts were referenced to illustrate each condition. These were mainly English language articles, but included appropriate non-English language papers. CONCLUSIONS: Balanitis is a common condition among genitourinary medicine clinic attendees, the cause often remaining undiagnosed. Many cases are caused by infection, with candida being the most frequently diagnosed. However, gardnerella and anaerobic infections are common, and there are a wide variety of other rarer infective causes. In addition irritant balanitis is probably a contributing factor in many cases. Balanitis which persists and in which the cause remains unclear warrants biopsy. PMID- 8707317 TI - The haemolytic uraemic syndrome in patients with AIDS. AB - OBJECTIVE: Thrombotic microangiopathies have been increasingly recognised in HIV infection. The contribution of haemolytic uraemic syndrome (HUS) has not received as much emphasis as other members of the thrombotic microangiopathies. We describe the clinical features and prognosis of HUS in a group of patients with AIDS. SETTING: St Bartholomew's and The Middlesex Hospitals, London. PATIENTS: Five HIV seropositive individuals with clinical and histological features consistent with HUS. INTERVENTIONS: Blood transfusion, fresh frozen plasma, haemodialysis, renal biopsy, autopsy. CONCLUSIONS: HUS occurs in advanced HIV infection. Hypertension was a prominent clinical feature in HUS in this patient group. Measures to limit renovascular damage were unsuccessful and haemodialysis was usually needed to support renal function. The prognosis is poor, no patient achieved clinical remission and all died. PMID- 8707316 TI - Sexually transmitted diseases in South Africa. AB - AIM: To review the epidemiology of and data collection for sexually transmitted diseases (STDs) in South Africa. METHODS: Literature published since 1980 on STDs in South Africa were complied and evaluated. Historical reports and salient unpublished literature were also used in the literature review. Studies were critically reviewed in the light of sample populations and study methods, and a baseline picture of the patterns of STD burden was developed. RESULTS: The STD burden in apparently asymptomatic study populations is significant. Ulcerative infections, primarily caused by syphilis and chancroid, are present in 5-15% of asymptomatic clinic attenders; prevalence rates of gonorrhoea average 8%, with up to 13% of gonococcal isolates resistant to penicillin antibiotics. In addition, on average, chlamydia and vaginal infections are detected in 16% and 20-49%, respectively, of antenatal and family planning clinic attenders. HIV seroprevalence rates have reached 7.6% in antenatal clinic attenders. Most South African STD data are derived from ad hoc surveys which have traditionally focused only on several major infections and particular urban centres. Almost all STD studies have been facility-based, with many studies based at STD clinics, thus reporting only relative frequencies and not population-based prevalences of STDs. With the possible exception of HIV, systematic surveillance data for STDs are conspicuously lacking. CONCLUSION: The disease burden of classic sexually transmitted infections has historically been heavy, and continues to be a serious public health problem in South Africa. Morbidity from both ulcerative and non ulcerative infections, particularly in women, is significant. The body of STD data, although mostly sound, remains incomplete, and with the rapid emergence of HIV in South Africa, surveillance of STDs and focused STD policies will be critical. PMID- 8707319 TI - The seed of an ulcer. PMID- 8707318 TI - Active syphilis in HIV infection: a multicentre retrospective survey. The German AIDS Study Group (GASG). AB - OBJECTIVE: To study syphilis in HIV infection focusing on immunocompromised patients with an atypical or aggressive clinical course of syphilis, inappropriate serological reactions or an unreliable response to therapy. STUDY DESIGN: A multicentre retrospective chart review using a standardised questionnaire for all patients with active syphilis. SETTINGS: Thirteen dermatological and medical centres throughout Germany, all members of the German AIDS Study Group (GASG). PATIENTS: Clinical data of 11,368 HIV infected patients have been analysed for cases of active syphilis requiring treatment. Asymptotic patients with reactive serological parameters indicating latent syphilis without a need for treatment were excluded. RESULTS: Active syphilis was reported in 151 of 11,368 HIV infected patients (1.33%, range per centre 0.3%-5.1%). Most of the 151 syphilis patients were male (93%) and belonged to the homosexual or bisexual exposure category for HIV infection (79%); another 6% were iv drug users. Among the 151 syphilis patients primary syphilis was diagnosed in 17.2%, maculopapular secondary syphilis in 29.1%, ulcerating secondary syphilis in 7.3%, neurosyphilis in 16.6% and latent seropositive syphilis without clinical symptoms but serological abnormalities indicating active syphilis in 25.2%. A history of prior treatments for syphilis was reported in 50%. At the time of syphilis diagnosis 26.5% of the patients were in CDC stage II, 33.8% in stage III and 24.5% in stage IV of HIV disease (CDC classification 1987). CD4 cell count was lowest in those with ulcerating secondary syphilis (mean 307, SD 140/microliters) and neurosyphilis (351, SD 235/ microliters). The highest CD4 count was found in patients with early primary and early secondary syphilis (444, SD 163/microliters and 470, SD 355/microliters). Inappropriate serological response to syphilis infection was found in 81 of 151 patients (54%). Remarkable findings were false negative VDRL titres (11 patients with non primary syphilis), false negative TPHA (1) or 19S-IgM-FTA-ABS-tests (16), and strongly reactive VDRL (> or = 512, 8) or TPHA titres (> or = 10 240, 47). Treatment failures were reported in at least 6 of 151 cases (4%). CONCLUSIONS: Atypical clinical and serological courses of syphilis were observed in HIV infected patients. Ulcerating secondary syphilis with general symptoms ("malignant syphilis") was 60 times more frequent than in historic syphilis series. Neurosyphilis was found in one sixth of those with active syphilis. Therefore lumbar puncture should be considered a routine in coinfections with HIV and syphilis. Treatment efficacy should be monitored carefully. PMID- 8707320 TI - Clinical algorithms for the screening of Chlamydia trachomatis in Turkish women. AB - OBJECTIVE: To test the diagnostic validity of clinical algorithms for the detection of Chlamydia trachomatis in an urban population of married women in Turkey. DESIGN: Cross-sectional population-based survey. SUBJECTS: A systematic sample of 867 women who reported the use of contraceptive methods. MAIN OUTCOME MEASURES: Sensitivity, specificity and positive predictive value of clinical algorithms for the diagnosis of C trachomatis. RESULTS: C trachomatis was diagnosed in 4.89% of the women. The WHO algorithm for use in settings where no vaginal examination could be performed had a sensitivity of 9% and a specificity of 96%. The corresponding figures for the WHO algorithm incorporating the findings of a speculum examination were 47% and 56% respectively. Algorithms incorporating symptoms or signs other than those suggested by the WHO did not yield satisfactory standards of validity. CONCLUSIONS: The findings of this study do not support the widespread introduction of the use of clinical decision models for screening of women for chlamydia infection in primary health care settings such as family planning or antenatal clinics. The large number of false positive results with the use of the clinical algorithms tested in this study would cause unnecessary costs to the health system and unnecessary interventions to the women treated. PMID- 8707321 TI - Occult miliary tuberculosis in advanced HIV disease. PMID- 8707322 TI - A case control study of HIV seroconversion in gay men, 1988-1993: what are the current risk factors? AB - OBJECTIVE: To investigate current risk factors for HIV seroconversion among homo/bisexual men. DESIGN: Matched case control study in a large STD clinic in central London. Data on risk factors were obtained by case note review. METHODS: Fifty-six cases who had at least one negative HIV test followed by a positive test between June 1988 and July 1993, and two homo/bisexual controls (having two or more negative HIV tests) matched to each case on age, total number of HIV tests and test interval period were identified. Univariate and multivariate odds ratio were calculated for acute STD, ano-genital intercourse, condom use and HIV status of sexual partners. RESULTS: Adjusted odds ratios (95% confidence intervals) for HIV seroconversion were 4.1 (1.3-13.3) for having an acute interval STD and 4.6 (1.4-15.4) for having a known HIV infected sexual partner. Compared with men who always used condoms, odds ratios for men who sometimes or never used condoms were 7.9 (2.2-28.9) and 16.2 (3.0-86.0) respectively. Unprotected ano-genital intercourse was commonly reported by both cases and controls, and reported condom use was no greater with a known HIV infected partner than with a partner of unknown HIV status. CONCLUSION: HIV seroconversion among homo/bisexual men attending STD clinics is strongly related to having an acute STD, a known HIV infected partner and not using condoms. Although consistent use of condoms is highly protective, knowing that a partner is HIV infected does not ensure condom use between serodiscordant men. More effective, well-evaluated interventions are needed to reduce sexual risk-taking in this population. PMID- 8707323 TI - Who goes to sexually transmitted disease clinics? Results from a national population survey. AB - OBJECTIVES: To examine the pattern of attendance at sexually transmitted disease (STD) clinics in Britain. To compare the demographic characteristics, behaviours and attitudes of STD clinic attenders with those of non-attenders, and to assess the extent to which STD clinics are used by those with high-risk sexual lifestyles. DESIGN: Random sample general population survey of sexual attitudes and lifestyle. SUBJECTS: 18,876 randomly selected men and women resident in Britain aged 16-59 years. MAIN OUTCOME MEASURES: Demographic characteristics, pattern of homosexual partnerships, heterosexual partnerships, payment for sex, abortion, drug injection in the last five years, and attitudes to sexual behaviours amongst 512 respondents who had attended STD clinic in the last five years compared with those who had not. RESULTS: 8.3% of men and 5.6% of women had attended a clinic in their lifetime and 3.4% and 2.6% respectively in the last five years. Attendance rates varied substantially with area of residence. 11% of Inner London residents had attended in the last five years. In multivariate analysis, STD clinic attendance for men was most strongly associated with increased numbers of heterosexual partners, (OR = 6.01 (4.44-8.15) and homosexual partnerships (OR = 9.59 (5.83-15.8)) and more weakly associated with payment for sex, non-manual social class, age 25-44, unmarried status and smoking. Clinic attendance for women was most strongly associated with numbers of heterosexual partners (OR = 3.74 (2.76-5.08) and injecting drug use (OR = 4.39 (1.73-11.1)). A weaker independent association was found with a history of abortion, anal sex, non-manual social class, non-married status and age 16-24. From the total population, 1 in 6 men and 1 in 7 women in the top 5% of the distribution for numbers of heterosexual partners and 1 in 5 men paying for sex and 1 in 4 of those with a homosexual partner had attended a clinic in the last five years. The probability of attendance increased with multiple risk behaviours. Of women 64.2% and of men 69.7% attending clinics reported major risk markers for STD transmission. CONCLUSIONS: STD clinics in Britain are used by a wide demographic spectrum of the population. The behaviours, but not the attitudes, of attenders differed markedly from those of non-attenders. Clinics are relatively efficient in attracting only those with high-risk lifestyles, but, at a population level, the minority of those reporting risk-markers for STD transmission attend clinics. These findings suggest that STD clinics are an important focus for sexual health promotion, but that community programmes are also important for reaching non attenders. PMID- 8707324 TI - Epidemiology of genital chlamydial infections in patients with chlamydial conjunctivitis; a retrospective study. AB - OBJECTIVE: To determine how often chlamydial conjunctivitis is accompanied by a genital chlamydial infection and if there is a correlation between the dominant hand and the eye first infected. METHODS: We retrospectively studied the records of 65 patients with chlamydial conjunctivitis who were referred to the Outpatient Department of Sexually Transmitted Diseases (STD) of the University Hospital Rotterdam by ophthalmologists of the Eye Hospital Rotterdam. The patients have recently been asked by letter if they were left- or right-handed. RESULTS: Twenty of the 37 men (54%) had a positive chlamydial urethral culture. Seventy per cent of these men had no genital symptoms. Eight of the 37 men (22%) had a non specific urethritis (NSU). Twenty of the 27 women examined (74%) had a positive chlamydial cervical culture. Sixty per cent of these women had no genital symptoms. Eight women with a genital chlamydial infection also had another genital infection. Five women without a genital chlamydial infection had another genital infection. Two women had no genital infection at all. A correlation between the eye infected and left- or right-handedness of the patient could not be found. CONCLUSIONS: A considerable percentage of the patients with a chlamydial conjunctivitis had a concomitant genital chlamydial infection. The majority of them had no genital symptoms. Since patients with chlamydial conjunctivitis and/or their partners possibly have a concomitant genital chlamydial infection, we recommend referral of both patients and sexual partners to an STD clinic for routine examination and systemic treatment when indicated. PMID- 8707325 TI - A comparison of referral patterns and characteristics of patients with first episode symptomatic genital HSV-1 and HSV-2 infections in Sheffield. AB - OBJECTIVE: To ascertain factors associated with HSV-1 and HSV-2 isolates in patients attending a genitourinary medicine clinic with symptomatic first episode genital herpes (FEGH). DESIGN: Retrospective study. SUBJECTS: A total of 606 females and 333 males presenting with culture positive FEGH between 1990-94. SETTING: Department of Genitourinary Medicine, Royal Hallamshire Hospital, Sheffield, UK. METHODS: Group comparison of referral patterns, demographic data, prior and concurrent episodes of STD, recent partner change. RESULTS: HSV-1 infected patients of either sex were more likely to be general practitioner (GP) referred, to be white, and less likely to have had preceding STD episodes. Recent sexual partner change had occurred significantly more often in HSV-2 infected females but there was no similar difference between HSV-1 and HSV-2 infected males. CONCLUSION: The relative HSV-1:HSV-2 isolate ratio in FEGH is influenced by the referral patterns. HSV-1 isolates predominate in patients presenting to GPs who refer the patients to GUM clinics for accurate diagnosis, counselling, follow up and screening for other STDs. PMID- 8707326 TI - Hepatitis B vaccination schedules in genitourinary medicine clinics. AB - OBJECTIVES: To compare two vaccination schedules in delivering hepatitis B vaccine to at-risk genitourinary medicine clinic attenders. SETTING: Genitourinary medicine clinic of St Thomas' Hospital, London, UK. METHODS: Two vaccination protocols were compared. Between January 1991 and December 1992, individuals had doses of recombinant hepatitis B vaccine at 0, 1 and 6 months (standard). From January until October 1993 doses of vaccine were administered at 0, 1 and 2 months (accelerated), following which timing of a booster dose was made on the basis of hepatitis B surface antibody (anti-HBs) assessment. Case notes were reviewed with regard to compliance rates and anti-HBs levels. RESULTS: Two hundred and fourteen individuals were included (standard 104, accelerated 110). Of the standard group 80.8% and 61.5% attended for the 2nd and 3rd doses respectively compared with 80.0% and 75.5% of the accelerated group (attendance for the 3rd dose chi 2 = 4.19, p < 0.05). For both of these groups compliance was significantly better in those who requested vaccination rather than being offered it opportunistically (chi 2 = 4.86, p < 0.05). Seroconversion rates were not significantly different between the two groups (87.5% versus 83.1%). A significantly higher proportion of the standard group, however, achieved anti-HBs levels greater than 100 i.u./l. CONCLUSIONS: Completion of hepatitis B vaccination was improved by changing to a 0, 1 and 2 month protocol. Levels of anti-HBs achieved in the accelerated group, however, were lower. If it is confirmed that maintaining anti-HBs levels is not important in retaining protection against hepatitis B then the accelerated schedule has clear advantage. If not, the advantages may be nullified by the need, in some, for an early booster. PMID- 8707327 TI - Hepatitis C virus infection in a male homosexual cohort: risk factor analysis. AB - BACKGROUND: Hepatitis C virus (HCV) infection is a major cause of morbidity throughout the world. Parenteral exposure to infected blood accounts for the majority of cases. Sexual transmission is suggested by the higher prevalence of infection in sex workers and homosexual men. Sexual practices which contribute to HCV infection need to be identified. METHODS: The social and medical history, and HCV serostatus of 1058 homosexual men in the Pittsburgh arm of the Multicenter AIDS Cohort Study were analysed. Multivariate analysis was used to determine risk factors for HCV seropositivity. RESULTS: 31 men were HCV seropositive by enzyme immunoassay and recombinant immunoblot assay (2.9%). They were more likely to be HIV seropositive (39%) than the HCV seronegative men (19%). Needle sharing and illegal drug use were the most important risk factors for HCV seropositivity. Statistically significant sexual factors (p < 0.05) included a history of syphilis, rectal gonorrhea, anal insertive intercourse with ejaculation, and douche or enema use before anal receptive intercourse. The number of sexual partners was not a significant risk factor. CONCLUSIONS: HCV infection is associated with specific sexually transmitted diseases (STDs) and sexual practices in the male homosexual population. The evidence of high risk behavior should be incorporated into ongoing educational efforts to decrease the incidence of STDs. PMID- 8707328 TI - Emergency hormonal contraception usage in genitourinary medicine clinic attenders. AB - OBJECTIVE: To assess the indications for usage of emergency hormonal contraception amongst a population of London genitourinary medicine clinic attenders. METHODS: In a prospective study, 150 consecutive women receiving emergency hormonal contraception (EHC) were enrolled. The attending doctor completed a questionnaire of patient details and prescribed EHC with prophylactic prochlorperazine. Follow-up was arranged three weeks later, at which time outcomes and side-effects of therapy were recorded. For those women who did not reattended as planned case notes were reviewed at three months. RESULTS: Of 150 women surveyed, 100 (66%) reported contraceptive method failure, 48 (32%) had used no contraception at the time of last sexual intercourse and two requested EHC after sexual assault. Ninety three (62%) reported condom failure, 7 (5%) oral contraceptive pill failure. Seventy five (50%) had used EHC before (range 1-10 times). Seventy one (47%) women reattended within three months. Five (3.3%) of the 150 women were pregnant; none of these cases had experienced nausea or vomiting whilst taking EHC. Side-effects were reported by 22 (31%) of the 71 patients who reattended. Nine (6%) women had been followed-up in the family planning advisory clinic. Of the 71 women who reattended, 39 (55%) reported that their preferred future method of contraception would be condoms. Of the 150 women 19 (13%) underwent tests for sexually transmissible infections within one month of presentation. CONCLUSIONS: EHC usage in this population was associated with a failure rate of at least 3.3% and an overall side effect rate of 31%. Despite requests for emergency contraception because of condom failure many elected to continue using condoms as their preferred method of contraception. The majority of women (53%) did not return for follow-up or family planning advice, and so we believe that future contraceptive plans must be addressed at the time EHC is prescribed. PMID- 8707329 TI - Why do men with urethritis in Cameroon prefer to seek care in the informal health sector? AB - OBJECTIVE: To study the impact of cost of treatment for sexually transmitted diseases (STDs) on the preference of men with urethritis to seek care in the informal sector. SUBJECTS AND METHODS: A random sample of young men from the general population of Yaounde and Douala, Cameroon, and of employees of a Yaounde factory underwent a structured interview on the occurrence of STD-related complaints and health seeking behaviour. Men who consulted in the formal sector were compared with men consulting in the informal sector, with regards to cost of medication and level of education. RESULTS: Sixteen percent of men in the general population and 20% of factory employees reported a history of urethritis in the 12 months preceding the interview. Of the men in the general population 53% had sought care in the formal sector; among the factory employees this proportion was 56%. Men who did not consult in the formal sector spent significantly less on drugs than men who did consult in the formal sector: median cost $14.4 versus $24.0 (p = 0.02) for drugs purchased in pharmacies; median cost $8 versus $32 for drugs purchased in small stores or from acquaintances. Preference for formal health services was associated with higher educational attainment. CONCLUSION: In large towns in Cameroon the utilisation of formal health services for STD related complaints is low and the high cost of treatment in the formal sector may play an important role in the choice of care option. PMID- 8707330 TI - An interview based approach to seeking user views in genitourinary medicine. AB - OBJECTIVES: To assess user and potential user views on the appropriateness, nature and quality of genitourinary medicine (GUM) provision in Bristol, UK and to develop a model for ongoing user consultation by GUM providers and purchasers. DESIGN: This qualitative study was based on semi-structured interviews with service users, potential users, community informants and NHS professionals. PARTICIPANTS: 76 current, past or potential users, 10 community informants and 11 NHS professionals were interviewed. African-Caribbean women and men, homeless men and women, lesbians and gay men, men and women living with HIV and women working in the sex industry were recruited to maximise the diversity of the sample. RESULTS: The interviews demonstrated that participants commented positively on many aspects of the service available. The research also identified a number of areas where the service could be improved. Many users emphasised their initial difficulty in finding out about the department and the need for greater publicity and outreach. Users reported coming to the clinic with high levels of anxiety and negative preconceptions about the GUM service. Specific issues were identified for different groups of users. There was a strongly expressed need from a number of women and African-Caribbean men for completely single sex clinics. CONCLUSIONS: The research identified a number of issues of importance to service users that had not been identified in the department's own questionnaire surveys. The results support the premise that qualitative interviews can be successfully employed to access a diverse sample of users, and can offer insights significantly beyond those available from structured patient questionnaires. PMID- 8707331 TI - Does outreach work help in the control of sexually transmitted disease? PMID- 8707332 TI - The glittering vagina--a seasonal phenomenon? PMID- 8707333 TI - Molecular mechanism of T-cell control of Chlamydia in mice: role of nitric oxide in vivo. AB - T-cell-mediated immunity is crucial for the control of Chlamydia in mice. Recent evidence from studies in an in vitro model of the mucosal epithelium, the polarized epithelial-lymphocyte co-culture (PELC) system, indicated that protective murine T cells mediated intracellular inhibition of the Chlamydia trachomatis agent of mouse pneumonitis (MoPn) at least partly by activating the interferon-gamma (IFN-gamma)-inducible nitric oxide synthase (iNOS) pathway. To investigate whether nitric oxide played a role in controlling chlamydial infection in vivo, the protective capacity of a chlamydial-specific T-cell clone (clone 2.14-0) was analysed in mice in the presence of a specific inhibitor of iNOS. The results revealed that the ability of this clone to clear Chlamydia in vivo is in part mediated by induction of nitric oxide (NO) production. The L arginine analogue and iNOS inhibitor, NG-monomethyl-L-arginine monoacetate (MLA), increased the chlamydial burden in infected mice and inhibited the ability of clone 2.14-0 to clear genital MoPn infection in vivo. The results are consistent with the working hypothesis that the IFN-gamma-inducible iNOS pathway is involved in the control of Chlamydia by T lymphocytes in mice. PMID- 8707334 TI - Expression of beta 2-microglobulin-free HLA class I alpha-chains on activated T cells requires internalization of HLA class I heterodimers. AB - HLA class I molecules on activated T cells are expressed as heterodimers associated with beta 2-microglobulin (beta 2-m) and also beta 2-m-free HLA class I alpha-chains. Mechanisms leading to the expression of the activation associated beta 2-m-free HLA class I alpha-chains are poorly defined, however. Upon enzymatical removal of HLA class I alpha-chains on activated T cells, re expression is observed within minutes upon reculture, reaching half-maximal levels within 1 hr. This process is independent of de novo protein synthesis and of export of newly synthesized proteins. Inhibition of the formation of coated pits by potassium depletion of cells abrogated the re-expression of HLA class I alpha-chains, suggesting that recycling events of HLA class I heterodimers via endosomal compartments are required for the generation of monoclonal antibody LA45-reactive alpha-chains. Furthermore, the rate of alpha-chain generation seems to be governed by the amount of cell surface-expressed HLA class I heterodimers. Taken together these findings suggest that beta 2-m-free HLA class I alpha-chains are generated during the process of class I heterodimer recycling. PMID- 8707335 TI - CD31 (PECAM-1) is a differentiation antigen lost during human CD4 T-cell maturation into Th1 or Th2 effector cells. AB - The CD31 antigen (PECAM-1) has been reported to be a stable marker for a human CD4 T-cell subpopulation unable to produce interleukin-4 (IL-4). We show here that CD31 expression is not stable inasmuch as CD4 T-cell lines and clones derived from cell-sorted neonatal CD31+ cells lose CD31 upon repetitive cycles of stimulation and IL-2 expansion. Moreover, various cytokines (IL-1 alpha, IL-4, IL 6, transforming growth factor-beta) fail to reinduce CD31 on CD31- clones. Whereas all CD31+ CD4 T cells rapidly express high levels of the CD45RO antigen and down-regulate the L-selectin antigen after priming, CD31 disappears more slowly because only part of the cells lose CD31 expression upon each cycle of stimulation. Loss of CD31 reflects a functional maturation of CD45RO+ cells since, in a system which favours the development of Th2 effectors, IL-4 is produced by CD31- but not CD31+ effector T cells, whereas interferon-gamma is produced by both types of cells. However, CD31 is not a Th1 marker since it is not expressed on several Th1 antigen-specific clones. We conclude that CD31 is a maturation marker expressed on the great majority of naive CD45RO- CD4 T cells and on a subset of CD45RO+ CD4 T cells that are at an intermediate stage of maturation. PMID- 8707336 TI - Modulation of WC1, a lineage-specific cell surface molecule of gamma/delta T cells augments cellular proliferation. AB - WC1, also known as T19, is the only unique gamma/delta T-cell differentiation antigen described to date other than the gamma/delta T-cell receptor. We present evidence that modulation of WC1 results in augmented proliferation of gamma/delta T cells. Immobilized IL-A29, a monoclonal antibody (mAb) specific for WC1, augmented proliferation of gamma/delta T cells in the autologous mixed leucocyte reaction (AMLR) as well as proliferation induced by either anti-CD3 or anti-CD5 mAb. In contrast, anti-CD5 mAb did not increase proliferation in the AMLR even though both CD5 and WC1 are members of the scavenger receptor cysteine-rich family of proteins and are expressed by bovine peripheral blood gamma/delta T cells. IL-A29 did not induce proliferation when assessed alone or in the presence of either phorbol myristate acetate (PMA) or interleukin-2. IL-A29 also did not induce detectable calcium mobilization when evaluated in the presence of monocytes, PMA, or following cross-linking of IL-A29 with anti-immunoglobulin antibody. We conclude that WC1 is a gamma/delta T-cell lineage-specific cell surface differentiation antigen which is involved in activation of gamma/delta T cells using an as yet unidentified pathway. PMID- 8707338 TI - Repopulation of blood lymphocyte sub-populations in rheumatoid arthritis patients treated with the depleting humanized monoclonal antibody, CAMPATH-1H. AB - Patients with severe rheumatoid arthritis who had failed treatment with conventional therapies were treated with a course of five or 10 daily intravenous infusions of CAMPATH-1H, a humanized antibody against the CD52 antigen, resulting in profound depletion of peripheral blood mononuclear cells. During the subsequent 18 months, lymphocytes were analysed for sub-populations by fluorescence-activated cell sorter (FACS) and for proliferation in response to polyclonal T-cell stimulation with anti-CD3 or staphylococcal enterotoxin B (SEB). Treatment resulted in almost complete depletion of lymphocytes from the blood followed by gradual repopulation. CD16+ natural killer (NK) cells and CD14+ monocytes returned to pretreatment levels within 1-2 months. CD19+ B cells returned to within 50% of pre-treatment levels by day 66 and to within normal range by day 150, whereas CD8+ T cells recovered to 50% of pretreatment levels by day 66, but did not show any further increase during the rest of the study period. The most profound effects were on the CD4+ T lymphocyte sub-population, as the mean CD4+ count did not increase above 20% of pre-treatment level at any time during the study period (550 days), at all the doses tested. The T cells which initially repopulated the blood 1-2 months after treatment, nearly all expressed the activation markers human leucocyte antigen (HLA)-DR and CD45RO, although the percentage of T cells expressing these molecules gradually declined to normal levels over time. Proliferative responses to polyclonal T-cell stimulation (anti-CD3 and SEB) were also significantly reduced in the first few months after treatment, but recovered to pre-treatment levels by day 250. The relationship between these observations and the clinical response is discussed. PMID- 8707337 TI - Impact of donor MHC class I or class II antigen deficiency on first- and second set rejection of mouse heart or liver allografts. AB - The influence of donor major histocompatibility complex (MHC) class I- or class II-deficiency on the initiation of first- and second-set rejection of mouse heart and liver allografts was examined. C3H (H-2k) mice received heterotopic cardiac or orthotopic liver grafts from unmodified B10 (H-2b), B6 (H-2b), b2m (H-2b; class I deficient) or AB0 (H-2b; class II deficient) donors. Organ survival was also investigated in C3H recipients that had been presensitized by a normal B10 skin graft 2-3 weeks before heart or liver transplantation. The absence of cell surface MHC class I or class II resulted in significant prolongation of primary cardiac allograft survival. Three of seven (43%) MHC class I-deficient, and two of five (40%) class II-deficient heart grafts were accepted indefinitely (survival time > 100 days). Thus both MHC class I and class II molecules appear to be important for the elicitation of first-set rejection in the heart allograft model. All liver allografts survived > 100 days in normal recipients. In C3H recipients that had been presensitized by a B10 skin graft, however, both heart and liver grafts from AB0 (class II deficient) donors underwent accelerated rejection (median survival time [MST] 3 and 4 days, respectively). In contrast, liver grafts from class I-deficient mice (b2m) were still accepted indefinitely by B10 skin-presensitized C3H recipients, whereas class I-deficient hearts survived significantly longer than those from class II-deficient or normal donors. These data demonstrate that the expression of donor MHC class I, and not class II is crucial in initiating second-set organ allograft rejection. In vitro monitoring revealed that at the time of organ transplant, both splenocytes and serum of the skin-presensitized animals displayed high cytotoxicity against AB0 (class II-deficient) but not against b2m (class I-deficient) targets. PMID- 8707339 TI - Late-term fetal thymectomy does not prevent the development of gut-homing T cells after birth. AB - Tissue-specific circulation of T cells is a critical element in the integration of systemic immune responses. Current models of T-cell migration suggest that homing specificities of T cells for tissues such as gut and skin are generated outside the thymus as a result of activation of virgin T cells by antigen in lymph nodes. We have used the sheep fetus (which is immunologically virgin and contains no memory or effector T-cell subsets) to examine the migration of 51Cr labelled T cells in vivo. We report that gut-homing T cells are not present in the fetus and that gut-homing T cells from postnatal lambs home normally to fetal gut. Fetal thymectomy performed immediately prior to birth failed to prevent the development of gut-homing T cells in postnatal life. Gut-homing specificities on T cells are thus acquired extrathymically. PMID- 8707340 TI - Chemical carcinogens and antigens induce immune suppression via Langerhans' cell depletion. AB - The ability of the chemical carcinogen dimethylbenz(a)anthracene (DMBA) to deplete Langerhans' cells (LC) from murine skin is crucial to the development of antigen-specific suppression. This depletion is a consequence of the LC recognizing the DMBA as antigenic and migrating to the draining lymph nodes to attempt to elicit T-cell activation. This depletion also occurred following exposure to high doses of the contact sensitizers 2,4-dinitrofluorobenzene (DNFB), 2,4,6-trinitrochlorobenzene (TNCB) and fluorescein isothiocyanate (FITC). However, LC depletion was not significant at lower doses, even though these doses were sufficient to induce strong contact sensitivity responses. Application of the contact sensitizer, DNFB, through skin depleted of LC (by pretreatment with either the carcinogen DMBA or the antigen TNCB) failed to induce contact sensitivity. This immune non-responsiveness was antigen specific, and could be transferred by spleen cells to naive mice, which were unable to respond to DNFB. Mouse skin treated with doses of TNCB, that did not cause LC depletion but still induced a normal contact hypersensitivity, retained its ability to initiate a normal immune response to DNFB. Together these findings demonstrate that carcinogens share some properties with antigens as they both cause LC depletion and interact with the immune system. Furthermore, it is this LC depletion, rather than carcinogen treatment, that is a critical factor which leaves the skin immunologically compromised and favours the induction of antigen-specific suppression. PMID- 8707341 TI - Catecholamines are synthesized by mouse lymphocytes and regulate function of these cells by induction of apoptosis. AB - The immune and the nervous systems are anatomically closely related and interact with each other by molecules common to both systems, such as cytokines and neurotransmitters. The purpose of this study was to investigate the participation of catecholamines in the neuroimmunological network. The ability of immune cells to produce catecholamines was examined by a highly sensitive capillary electrophoresis assay, which permits detection of easily oxidized catecholamines in the zeptomole (10(-21)) range. In addition, the effects of catecholamines on in vitro proliferation, differentiation and apoptosis of lymphocytes were assessed. Mouse spleen cells and macrophages contained on average 7 x 10(-17) and 2 x 10(-17) mole dopamine per cell, respectively. In the former cell population also norepinephrine was found. Several mouse B- and T-cell hybridomas were also shown to contain endogenously produced dopamine in levels ranging from 7 x 10( 20) to 2 x 10(-18) mole dopamine per cell. In addition, one of the T-cell hybridomas proved to synthesize norepinephrine. The dopamine production of lymphocytes was blocked by the tyrosine hydroxylase inhibitor alpha-methyl-p tyrosine, whereas incubation with the precursor L-DOPA increased the dopamine content. Incubation with L-DOPA, dopamine and norepinephrine dose-dependently suppressed mitogen induced proliferation and differentiation of mouse lymphocytes. Even short-time pretreatment of lymphocytes with L-DOPA and dopamine strongly suppressed lymphocyte proliferation and cytokine production. Incubation of lymphoid cells with L-DOPA, dopamine and norepinephrine dose-dependently induced apoptosis which, at least partly, explains the suppressive effects of catecholamines on lymphocyte function. Our results demonstrate that catecholamines: (i) are actively produced by lymphocytes and (ii) have the capacity to act as auto- and/or paracrine regulators of lymphocyte activity through induction of apoptosis. PMID- 8707342 TI - Antigen presentation in the murine oral epithelium. AB - We have previously reported that the buccal mucosa can support delayed type hypersensitivity (DTH) reactions to contact sensitizers. In the present study, we show that cells isolated from the buccal epithelium are able to present soluble exogenous antigens to specific T cells. Single cell suspensions obtained by enzymatic dispersion of buccal epithelial sheets could present the native protein antigen hen-egg lysozyme (HEL) to the I-Ak-restricted CD4+ T-cell hybridoma specific for a.a 46-61 on HEL. T-cell activation resulted in interleukin-2 (IL-2) production which could be inhibited by anti-major histocompatibility complex (MHC) class-II antibodies of pertinent specificity. Immunohistochemical staining of whole buccal epithelial sheets revealed that all MHC II positive cells had a dendritic morphology and expressed ATPase activity, indicating that these cells represent a major antigen-presenting cell (APC) population in this tissue. Furthermore, single cell suspensions isolated from buccal epithelium (BEC) after local in vivo administration of either a native soluble protein, a synthetic dodecapeptide, or a contact sensitizer were able to activate antigen-specific T cells ex vivo. Kinetic analyses indicated that maximal APC activity in the oral epithelium occurred within 1 hr after local antigen administration, and had essentially vanished after 24 hr. Conversely, APC activity was undetectable in draining cervico-mandibular lymph node cell suspensions recovered 1 hr after local antigen injection but became manifest after 3-24 hr. These observations suggest that dendritic cells can acquire antigens in the buccal epithelium and migrate to draining lymph nodes where they present processed antigen to MHC class II-restricted T cells. This APC population may thus be a critical element in the initiation of Th1-driven DTH responses in the oral mucosa. PMID- 8707343 TI - Expression and function of membrane regulators of complement on rat astrocytes in culture. AB - Human astrocytes express CD59, decay accelerating factor and membrane cofactor protein to restrict the damaging effect of complement (C) activation on their cell surface. 5I2 antigen (5I2 Ag) is the functional analogue of the latter two proteins in rats. We here demonstrate the surface expression on rat astrocytes of CD59 and 5I2 Ag and use sodium dodecyl sulphate-polyacrylamide gel electrophoresis and Western blotting to confirm their identity and to quantify expression. Rat CD59 (MW 20,000) was expressed at 720 x 10(3) molecules per cell and 5I2 Ag (MW 58,000 and 64,000) at 625 x 10(3) molecules per cell. Reverse transcription-polymerase chain reaction using specific oligonucleotide primers demonstrated expression of mRNA for each protein. Twenty-four-hour stimulation with inflammatory cytokines (interferon-gamma, tumour necrosis factor-alpha, interleukins-1 beta, -2 and -6) or phorbol myristate acetate had no significant effect on the level of expression of either protein as determined by Western blotting. Lysis caused by classical pathway activation of C in human or rat serum was enhanced by blocking the function of CD59 and 5I2 Ag on rat astrocytes with monoclonal antibodies. PMID- 8707344 TI - Novel polymer-grafted starch microparticles for mucosal delivery of vaccines. AB - Recent studies have demonstrated that systemic and mucosal administration of soluble antigens in biodegradable microparticles can potentiate antigen-specific humoral and cellular immune responses. However, current microparticle formulations are not adequate for all vaccine antigens, necessitating the further development of microparticle carrier systems. In this study, we developed a novel microparticle fabrication technique in which human serum albumin (HSA) was entrapped in starch microparticles grafted with 3-(triethoxysilyl)-propyl terminated polydimethylsiloxane (TS-PDMS), a biocompatible silicone polymer. The immunogenicity of HSA was preserved during the microparticle fabrication process. Following intraperitoneal immunization of mice, TS-PDMS-grafted microparticles (MP) dramatically enhanced serum IgG responses compared with ungrafted MP and soluble HSA alone (P < 0.001). When delivered orally, both TS-PDMS-grafted and ungrafted microparticles elicited HSA-specific IgA responses in gut secretions, in contrast to orally administered soluble antigen. Indeed, TS-PDMS-grafted microparticles stimulated significantly stronger serum IgG (P < 0.005) and IgA (P < 0.001) responses compared with those elicited by ungrafted microparticles. These findings indicate that TS-PDMS-grafted starch microparticles have potential as systemic and mucosal vaccine delivery vehicles. PMID- 8707345 TI - Ability of spleen, peritoneal cavity, and lymph node B cells to reconstitute serum immunoglobulin in SCID mice. AB - The impact of intrinsic B lymphocyte heterogeneity and of microenvironmental influences on serum immunoglobulin production by B cells was examined by intravenous (i.v.) and intraperitoneal (i.p.) transfer of BALB/c and BALB.xid (X chromosome-linked immunedefective; XID) lymph node (LN), splenic (SP) and peritoneal cavity (PerC) cells into severe-combined immune-defective (SCID) mice. The results indicate that each B-cell source restores all immunoglobulin classes within 5 weeks of transfer, the rates for each isotype, however, differ between the B-cell sources. Serum IgM levels were restored most rapidly by PerC cell transfer, followed by SP and LN cell transfer. In addition, normal immunoglobulin levels were reached in the absence of complete lymphoid reconstitution. Serum immunoglobulin phenotypes characteristic of the donor strain, e.g. reduced IgM and IgG3 production by XID B cells, were maintained after transfer into the SCID recipient. Microenvironmental influences were indicated by reduced immunoglobulin production after i.p. transfer and after i.v. transfer into irradiated SCID recipients. The data show that both B-cell type and microenvironment play significant roles in generating the heterogeneous pool of B cells required for humoral immunity. PMID- 8707346 TI - Fetal lambs are depleted of IgM+ cells following a single injection of an anti IgM antibody early in gestation. AB - B-cell depleted fetal sheep were created following a single injection of an anti IgM monoclonal antibody early in gestation. Six sheep fetuses were given a single intraperitoneal injection of a monoclonal antibody directed against IgM at 63 days of gestation (gestation in sheep = 150 days). The fetuses were killed at 138 142 days of gestation and lymphoid tissues were collected for subsequent light microscopy and immunohistochemical examination. The ileal and jejunal Peyer's patch (PP) follicles in four of the six injected fetuses were markedly reduced in size. Cells in the rudimentary follicles of the ileal PP of these animals showed no reactivity for IgM and most were negative for CD45. The dome regions contained many T cells, which were predominantly CD8+ cells and included gamma delta T cells. The interfollicular areas of the PP of the markedly affected fetuses contained large populations of T cells. The spleen and lymph nodes were also markedly depleted of IgM+ cells and these tissues contained only a small, scattered population of weakly IgM+ cells. Follicular accumulations of IgM+ cells were absent. Large populations of T cells were present in the white pulp of the spleen and cortex of the lymph nodes. The liver did not contain IgM+ cells and the medulla of the thymus was depleted of IgM+ cells. The results of this study suggest that a surface IgM+ B-cell population is present in the sheep fetus at 63 days of gestation, which is essential for the colonization of the ileal PP and subsequent B-cell development. PMID- 8707347 TI - The Epstein-Barr virus-binding site on CD21 is involved in CD23 binding and interleukin-4-induced IgE and IgG4 production by human B cells. AB - Human CD21 has previously been described as a receptor for the C3d,g and iC3b proteins of complement, as a receptor for the gp350/220 envelope glycoprotein of the Epstein-Barr virus (EBV) and also as a receptor for inerferon-alpha (IFN alpha). Structurally, CD21 consists of 15 to 16 short consensus repeats (SCR) of 60 to 75 amino acids followed by a transmembrane domain and an intracytoplasmic region. We reported that CD23, a low-affinity receptor for IgE (Fc epsilon R2), is a new functional ligand for CD21. We recently found that the sites of interaction of CD23 on CD21 are on SCR 5 to 8 and 1-2. The first site is a lectin sugar type of interaction and the second site is a protein-protein interaction. We report here that amongst the other ligands for CD21 (EBV, C3d,g and IFN alpha), only EBV is able to inhibit the binding of CD23 to CD21. Furthermore, even a peptide from gp350/220 of EBV known to bind to CD21 is able to decrease CD23 binding to CD21. Since CD23/CD21 pairing is important in the control of IgE production, we tested the effect of the EBV-derived peptide on immunoglobulin production from peripheral blood mononuclear cells and purified tonsillar B cells. Interestingly, the EBV-peptide inhibited IgE and IgG4 production induced by interleukin-4, in a dose-dependent manner. The same results were obtained using either peripheral blood mononuclear cells or purified tonsillar B cells. Another CD21 ligand, C3, did not affect binding of CD23 to CD21 nor the production of IgE and IgG4. This study indicates that blocking CD23 binding to CD21 SCR 2 on human B cells selectively modulates immunoglobulin production. PMID- 8707348 TI - Role of interleukin-2 and interferon-gamma in inducing production of IgG subclasses in lymphocytes of human newborns. AB - Unlike lymphocytes from adults, lymphocytes from cord blood of neonates cannot synthesize immunoglobulin G (IgG) in response to pokeweed mitogen (PWM). By using this mitogen in concert with interferon-gamma (IFN-gamma), interleukin-2 (IL-2), or interleukin-6 (IL-6), we studied the induction of IgG subclass molecules in lymphocytes of human neonates. IFN-gamma induced a limited, but substantial, enhancement of IgG2 production by neonatal lymphocytes. IL-2 dose dependently increased the production of each neonatal IgG subclass, whereas IL-6 did not. However, in adult lymphocytes, and under specific conditions, IL-6 or IL-2 each increased the production of all four IgG subclasses. Early in the culture IFN gamma synergized with IL-2 during the latter or whole culture period to enhance cord blood IgG2 levels. This finding contrasted with the adult IgG2 synthesis synergistically up-regulated by IFN-gamma and IL-6. IL-2 caused a graded increase in immunoglobulin production in neonatal lymphocytes with IgG3 being the highest and IgG2 the lowest, thus corresponding to the differential increase of serum levels of IgG3/IgG1 and IgG4/IgG2 early in childhood. Results suggest that IL-2, but not IL-6, is critical to the development of human IgG subclass production. PMID- 8707349 TI - Production of granulocyte-macrophage colony-stimulating factor by T cells is regulated by B7 and IL-1 beta. AB - Granulocyte-macrophage colony-stimulating factor (GM-CSF) has proliferation- and differentiation-inducing effects on immature myeloid cells in the bone marrow, and it can modulate the function of several types of mature myeloid cells. We have stimulated purified human T cells with immobilized anti-CD3 or mitogenic anti-CD2 (a combination of monoclonal antibodies 9-1 and 9.6) which could induce GM-CSF production. The cytokines interleukin-1 beta (IL-1 beta) and IL-2 strongly enhanced GM-CSF production, while IL-4, IL-6, GM-CSF, interferon-gamma (IFN gamma) and tumour necrosis factor (TNF) had no effect. Activation of protein kinase C by phorbol myristate acetate or triggering of CD28 on T cells by monoclonal antibody 9.3 provided accessory signals for enhanced GM-CSF production in activated T cells. Most important, the addition of mouse cells transfected with human B7-1 (CD80), a natural ligand for CD28, provided a potent accessory signal for GM-CSF production by activated T cells, which could not be blocked by cyclosporin A. The effect of IL-1 beta was in fact indirect, and resulted from enhanced IL-2 production, while the effect of B7 resulted from both IL-2 dependent and IL-2-independent pathways. We conclude that antigen-presenting cells (APC) can up-regulate GM-CSF production through IL-1 beta and through CD28 triggering by B7 molecules. As GM-CSF itself up-regulates B7 expression and IL-1 beta production by APC, a bidirectional regulatory feedback pathway between APC and T cells seems to modulate GM-CSF production. PMID- 8707350 TI - HIV-1 envelope glycoprotein gp120 down-regulates CD4 expression in primary human macrophages through induction of endogenous tumour necrosis factor-alpha. AB - Among immunological abnormalities present in human immunodeficiency virus type 1 (HIV-1)-infected individuals are dysregulation of cytokine production and CD4 down-regulation in both T-helper cells and monocytes/macrophages. The HIV-1 envelope glycoprotein 120 (gp120) has the ability to induce different cytokines in peripheral blood mononuclear cells and in monocytes/macrophages in vitro which in some instances have been reported to down-regulate macrophage CD4 expression. This study provides evidence that HIV-1 recombinant gp120 (rgp120) down-regulates both surface and total CD4 expression in primary tissue culture-differentiated macrophages (TCDM) at the level of transcription. The CD4 down-regulation observed in TCDM occurred between 6 and 12 hr after rgp120 treatment preceded by a peak of endogenous tumour necrosis factor-alpha (TNF-alpha) observed at 3-6 hr post-treatment. We demonstrate that the TCDM CD4 down-regulation observed after rgp120 treatment was inhibited by the use of an anti-huTNF-alpha monoclonal antibody (mAb), but not by mAb directed against other cytokines induced by rgp120, such as interleukin-1 beta (IL-1 beta) and interferon-alpha (IFN-alpha). The present findings roughly parallel those observed both in the sera of patients and in the monocytes/macrophages isolated from HIV-positive individuals, suggesting that gp120 by stimulating endogenous TNF-alpha production could be a good candidate for the CD4 down-regulation observed in the monocytes/macrophages of HIV-1-infected individuals. In contrast to CD4 down-regulation in HIV-infected lymphocytes, which results from a direct effect of viral genes on CD4 expression, soluble factors such as cytokines induced during HIV infection might explain the monocyte/macrophage CD4 dysregulation observed in acquired immune deficiency syndrome. PMID- 8707351 TI - Heat-shock protein expression on the membrane of T cells undergoing apoptosis. AB - Heat-shock proteins (hsp) represent a highly conserved family of proteins, normally localized in the cytoplasm and nucleus, whose expression is induced in situations involving cell stress. This paper reports the unusual translocation of hsp to the cell membrane of T cells undergoing apoptosis. We observed that glucocorticosteroid-induced thymocyte death is associated to the surface expression of hsp 60 and hsp 70 in a discrete fraction of apoptotic cells. hsp surface expression is closely related to a thymic subset of immature CD3low/- T cells. The expression of surface hsp 60 appears early after treatment with dexamethasone (3 hr) whereas the membrane expression of hsp 70 follows different kinetics and peaks later. Morphological analysis of the hsp+ apoptotic cells suggest that this subset represents late-stage apoptotic cells at their minimal volume before fragmentation into apoptotic bodies. Membrane expression of hsp is also associated with apoptosis in peripheral blood mononuclear cells from AIDS patients cultured in vitro. Altogether, we show that a discrete fraction of cells undergoing apoptosis expresses membrane hsp 60 and hsp 70, supporting the hypothesis that apoptosis causes a radical alteration in the expression of cell surface molecules. Surface hsp expressed during apoptosis may constitute a novel immune-context able to generate packages of self- and exogenous antigens, originating from degradation of altered cells. PMID- 8707352 TI - The role of tumour necrosis factor-alpha in combination with interferon-gamma or interleukin-1 in the induction of immunosuppressive macrophages because of Mycobacterium avium complex infection. AB - The role of some cytokines including tumour necrosis factor-alpha (TNF-alpha), interleukin-1 alpha (IL-1 alpha), interferon-gamma (IFN-gamma), transforming growth factor-beta (TGF-beta) and interleukin-6 (IL-6) in the generation of immunosuppressive macrophages (M phi s) in host spleen cells of Mycobacterium avium complex (MAC)-infected mice was studied. M phi populations with potent suppressor activity against concanavalin A (Con A)-induced mitogenesis of splenocytes (SPCs) were elicited not only in euthymic but also in athymic nude mice during MAC infection. The suppressor M phi s are, therefore, inducible not only through a T-cell-dependent mechanism but also through T-cell-independent mechanism. However, MAC-induced M phi s of athymic mice displayed about four times lower suppressor activity than those of euthymic mice, indicating that mature T cells are important for M phi activation to the highly immunosuppressive state. Anti-TNF, anti-IFN-gamma, and anti-TGF-beta antibodies (Abs) but not anti IL-6 Ab inhibited in vivo generation of MAC-induced immunosuppressive M phi s, and the neutralizing efficacy was in the order of anti-IFN-gamma Ab > anti-TNF Ab > anti-TGF-beta Ab. The effects of TNF-alpha, IL-1 alpha, IL-6, and IFN-gamma alone or combinations of them upon the acquisition of the suppressor activity by cultured splenic M phi s were studied. When normal splenic M phi s were treated with each cytokine for 3 days, TNF-alpha, IFN-gamma, and IL-1 alpha alone caused a slight elevation of their suppressive activity. Treatment of the normal M phi s with the combination of either TNF-alpha+IL-1 alpha or TNF-alpha+IFN-gamma yielded a marked increase in the suppressor activity, followed by IL-1 alpha+IFN gamma. These findings indicate the important roles of TNF-alpha, IFN-gamma, and IL-1 alpha in the generation of MAC-induced suppressor M phi s. PMID- 8707353 TI - Interferon-gamma differentially regulates antigen-processing functions in distinct endocytic compartments of macrophages with constitutive expression of class II major histocompatibility complex molecules. AB - RAW264.7 cells were transfected to express constitutively the murine class II major histocompatibility complex (MHC-II) molecule, I-Ak. The resulting RAW.Ak cells presented HEL(46-61) peptide to 3A9 T hybridoma cells, but they were unable to process and present HEL protein in their resting state. However, IFN-gamma stimulation induced the ability of RAW.Ak to process and present HEL protein, with little effect on their ability to present HEL(46-61) peptide. Antigen catabolism showed little change with IFN-gamma stimulation, suggesting that the production of peptides was not the regulated step in the processing pathway. Furthermore, HEL(46-61) peptide delivered directly into lysosomes by acid resistant liposomes was also presented only upon IFN-gamma stimulation, while the presentation of peptides delivered into endosomes by acid-sensitive liposomes showed a lesser dependence on IFN-gamma stimulation. Thus, IFN-gamma regulated the ability of peptides delivered into certain lysosomal compartments to meet with MHC-II molecules and form peptide-MHC complexes, or to transport subsequently to the plasma membrane. Two other antigens, ribonuclease A and haemoglobin, were processed by RAW.Ak cells without IFN-gamma stimulation, suggesting that these antigens could be processed by different mechanisms, perhaps in earlier endocytic compartments. Thus, different antigens may be processed in distinct endocytic compartments, and an IFN-gamma-regulated mechanism controls the rescue of peptides from lysosomal compartments for presentation at the plasma membrane. PMID- 8707354 TI - Thrombin stimulates production of interleukin-8 in human umbilical vein endothelial cells. AB - Interleukin-8 (IL-8) is regarded as an important mediator of inflammation because of its potent and specific chemotactic activity on neutrophils. In the present investigation, human umbilical vein endothelial cells (HUVEC) stimulated with thrombin were found to produce IL-8, in a dose- and time-dependent manner. After stimulation with 10 U/ml thrombin for 24 hr, the level of IL-8 in the conditioned medium was 14 ng/ml, or enough to elicit PMN chemotaxis in vitro. Northern blot analysis revealed that thrombin as well as IL-1 beta elevates the level of IL-8 mRNA preceding the formation of IL-8 protein. A synthetic peptide SFLLRN [human thrombin receptor-activating peptide (TRAP)] was found to mimic the action of thrombin. Preincubation with anti-thrombin compounds such as hirudin and antithrombin-III-heparin almost completely suppressed the action of thrombin without affecting the actions of other stimuli including IL-1 beta, phorbol 12 myristate 13-acetate (PMA) and TRAP. Diisopropylfluorophosphate-treated thrombin did not stimulate IL-8 production. Calphostin-C, a protein kinase C (PKC) inhibitor, attenuated the production of IL-8 by thrombin, TRAP and PMA, but left the action of IL-1 beta unchanged. These results strongly suggest that catalytic activation of thrombin receptor by thrombin results in PKC-dependent IL-8 production accompanied by an increase in IL-8 mRNA level. PMID- 8707356 TI - Mechanisms of pertussis toxin-induced myelomonocytic cell adhesion: role of Mac 1(CD11b/CD18) and urokinase receptor (CD87). AB - Stimulation of monoblastic U937 cells with transforming growth factor beta 1 and 1,25-(OH)2 vitamin D3 (TGF-beta 1/D3) upregulates urokinase receptor (uPAR) and confers urokinase-dependent adhesiveness to the cells for serum- or vitronectin coated surfaces. Recent studies show that uPAR itself is a high-affinity adhesion receptor for vitronectin and that urokinase (uPA) is an activator of this adhesive function. In the course of exploring possible G-protein involvement in this adhesion it was observed that TGF-beta 1/D3-primed U937 cells became adhesive to vitronectin in an uPAR-dependent manner when exposed to pertussis toxin (PTX). The adherent response is concentration- and time-dependent, and was not due to the ADP-ribosyltransferase activity of the toxin because the purified B-subunit of PTX was equally effective. Although promoting adhesion to serum- or vitronectin-coated surfaces, PTX blocked spontaneous cell adhesion to fibrinogen, an endogenous ligand for the Mac-1 receptor (CD11b/CD18). Flow cytometry study showed that expression of the alpha-subunit of Mac-1 (CD11b) on primed cells was increased by nearly threefold. Monoclonal antibody to CD11b abolished the PTX induced cell adhesion and the binding of the primed cells to PTX-coated plates. Activation of Mac-1 receptor by its endogenous ligand fibrinogen induced cell adherent response similar to PTX. PTX, but not uPA, triggered a rapid rise in [Ca2+]i in primed U937 cells, and PTX-induced adhesion was significantly attenuated by 1,2-bis-(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid/acetoxy methyl ester (BAPTA/AM), a selective membrane-permeant [Ca2+]i chelator. PTX induced cell adhesion was also prevented by antibodies to uPAR and by conditioned medium containing soluble uPAR. Together these data indicate that PTX B-subunit may bind to Mac-1 integrin, which leads to a rapid rise in [Ca2+]i and subsequent activation of uPAR for adherence to vitronectin, suggesting a functional link between Mac-1 and activation of uPAR important to cellular trafficking and host defence in response to Bordetella pertussis infection. PMID- 8707357 TI - Expression of major histocompatibility complex class II antigens on normal porcine intestinal endothelium. AB - A novel monoclonal antibody (MIL 11) specific for an antigen expressed on porcine endothelial cells is described. The antigen recognized by MIL 11 is most strongly expressed in the intestine but is also expressed on the capillary endothelium of a wide range of tissues. Using two- and three-colour immunofluorescence microscopy we demonstrated the extensive coexpression of MIL 11 and major histocompatibility complex (MHC) class II antigens on normal porcine capillary endothelium in the intestine, trachea, thymus and small veins, while endothelium of large vessels and the heart were negative for MHC class II. In contrast to humans and rodents, available reagents do not detect MHC class II on the intestinal epithelium of pigs. However, porcine intestinal endothelium expressed both DR and DQ antigens. A population of strongly class II-positive cells was also detected immediately adjacent to the endothelium in the lamina propria. Three-colour immunofluorescence microscopy highlighted the close association between endothelium and intestinal CD4+ T cells. Lamina propria T cells were mainly MHC class II positive, whereas those in the epithelial compartment were MHC class II negative. PMID- 8707355 TI - Interleukin-12 induces cytotoxic NK1+ alpha beta T cells in the lungs of euthymic and athymic mice. AB - We recently reported that interleukin-12 (IL-12) stimulated hepatic NK1.1 Ag+ alpha beta T cells with intermediate T-cell receptor (TCR; NK1+ TCRint cells) and enhanced their NK1 expression (NK1high TCRint), and that these cells acquire strong major histocompatibility complex (MHC) unrestricted cytotoxicity in C57BL/6 mice, both +/+ and nu/nu. In the present study, we find that although murine lung normally has few NK1+ TCRint cells, NK1high TCRint cells are induced in+/+ and nu/nu mice after systemic administration of IL-12; these cells exhibit strong MHC unrestricted cytotoxicity against NK-sensitive and -resistant targets. A small number of NK1high TCRint cells was also found in peripheral blood after increased amounts of IL-12 were administered. Cytotoxicity tests in vitro revealed that the cytotoxic activity of the lung mononuclear cells (MNC) of C57BL/6 mice induced by IL-12 was abrogated by the depletion of either NK1+ or CD3+ cells, but not of CD8+ cells, as reportedly was the case of hepatic MNC, suggesting that NK1high TCRint cells are an antimetastatic population not only in the liver but also in the lung of mice. IL-12 injection into mice markedly elevates serum interferon-gamma (IFN-gamma) levels. However, although IL-12 induced cytotoxicity of NK1high TCRint cells was significantly reduced by anti IFN-gamma antibody injection (which decreased serum IFN-gamma to an undetectable level), the appearance of NK1high TCRint cells in the lung and liver was not so affected. These results suggest that IFN-gamma is an important mediator of the cytotoxicity of NK1high TCRint cells but is not an essential factor for induction of these cells. We also added data showing that IL-12 has a broad antimetastatic effect against various liver and lung metastatic tumours intravenously injected into several strains of mice, including NK-deficient bg/bg mice. It can be considered that, in addition to NK cells, CD8+ cytotoxic T cells and gamma delta T cells, NK1+ TCRint cells can be categorized as one of the cytotoxic effector populations. These novel type cells distinct from regular T cells may play an important role in monitoring intra- and perivascular areas. PMID- 8707358 TI - Trend of increasing levels of minimum inhibitory concentration of ciprofloxacin to Salmonella typhi. AB - The minimum inhibitory concentration (MIC) of ciprofloxacin has been determined in 193 strains of S. typhi isolated during January and July each year from March 1991 to July 1995. While all strains remained within established range of susceptibility, a clear trend of increase of MIC values with time was observed. During the first four study periods, all strains had MIC values below 0.031 microgram/ml; during the fifth and sixth, 3 strains had a higher value namely 0.125 microgram/ml. During the last four, 14 strains showed MIC values of 0.25 microgram/ml. Thus an increase in MIC of ciprofloxacin to S. typhi has been observed over the years. PMID- 8707359 TI - Isolation of Mycobacterium tuberculosis from cerebrospinal fluid by the centrifugation & filtration methods. AB - Cerebrospinal fluid (CSF) samples were collected in 2 bottles each, from 112 children, examined clinically for tuberculous meningitis (TBM) One was processed by the centrifugation method and the other by the filtration method for the isolation of M. tuberculosis. Of these specimens, 11 and 13 yielded M. tuberculosis by the centrifugation method and the filtration method, respectively. In 7 specimens M. tuberculosis was isolated by both the methods; in 4, only by the centrifugation method, and in 6, only by the filtration method. Using both the methods, 17 (15.2%) of 112 specimens were culture positive for M. tuberculosis. The improvement in the rate of isolation, thus obtained, assumes importance as the confirmation of the diagnosis of TBM in all the clinically suspected cases is always desired. Moreover, the filtration method is simple and inexpensive and it can be carried out even in remote hospitals and the membranes, after filtration, can be transported to central mycobacteriology laboratory for culture of tubercle bacilli. PMID- 8707360 TI - Microbiological quality & incidence of organisms of public health importance in food & water in Ludhiana. AB - Bacteriological analysis of 713 samples of various types of foods and related articles and potable water samples from different places in Ludhiana, Punjab was carried out. The highest counts ranging from 2.5 x 10(6)-7.5 x 10(8) organisms/g were observed in raw vegetables and fruits, followed by 3 x 10(6)-9.8 x 10(7)/ml, 8.3 x 10(4)-8.9 x 10(7)/g and 1 x 10(3)-6.7 x 10(7)/g in fruit juice, milk and its products, and salty/non milk snacks respectively. Fresh chapati, dal, rice, cooked vegetables and karhi etc., showed no microbial contamination. However, samples of these articles from road side cafes gave counts up to 1 x 10(7) organism/g. The most probable number of coliforms and Escherichia coli/100 ml of water ranged from < 1 to > 1100. Although 1332 isolates of 16 types of organisms of public health significance were obtained those of proven enteropathogenicity were enterotoxigenic Esch. coli (55), Esch. coli O157 (3), enteropathogenic Esch. coli (1), enterotoxigenic Klebsiella (23), Streptococcus faecalis (152), Bacillus cereus (133), Staphylococcus aureus (125), Aeromonas spp (47), Salmonella spp (10), Shigella spp (4) and Yersinia enterocolitica (2). Poor quality of potable water and widespread occurrence of enteropathogens in food consumed by the common man in Ludhiana was evident. PMID- 8707361 TI - Comparison of regimens of amphotericin B deoxycholate in kala-azar. AB - A total of 288 parasitologically proved patients of kala-azar were randomly allocated to three treatment groups. Patients in groups A, B and C received amphotericin B (AMB) in a dose of 1 mg/kg body weight (bw)/day, 0.75 mg/kg bw/day and 0.5 mg/kg bw/day for 20 days respectively. Apparent cure (afebrile at the end of therapy) occurred in all patients and parasitological cure in 96 (100%), 92 (96%) and 84 (88%) patients respectively in groups A, B and C. Ultimate cure (no relapse in six months of follow up) occurred in 95 (99%), 87 (91%) and 79 (82%) patients in groups A, B and C respectively. The difference between the ultimate cure in the three groups was significant (P < 0.05). The incidence of adverse events (rise in serum creatinine and fall in serum potassium, loss of appetite and shivering, rigor and fever during infusion indicative of renal, GIT and infusion related toxicities respectively) was similar in the three groups. This study showed that amphotericin B should be given at a dosage of 1 mg/kg bw/day for 20 days for Indian kala-azar patients to minimise relapses and prevent development of drug unresponsiveness. PMID- 8707362 TI - A comparison of five staining methods for detection of Cryptosporidium oocysts in faecal specimens from the field. AB - Five staining methods for identification of Cryptosporidium oocysts in faecal smears were evaluated using samples obtained from asymptomatic south Indian villagers of all ages. The safranine-methylene blue technique was used as the gold standard and compared with 2 methods each using auramine and mepacrine, with potassium permanganate and carbol fuchsin as counterstains. All five methods identified all samples containing cryptosporidium oocysts, however false positive identifications were obtained by the standard auramine-phenol-potassium permanganate method. The auramine-carbol fuchsin and the mepacrine-carbol fuchsin techniques are the simplest and most rapid of the five methods compared. The mepacrine-carbol fuchsin technique is recommended for screening large numbers of stools, since it does not stain yeasts, is cheaper and less toxic than auramine. PMID- 8707363 TI - Molecular abnormalities of bcr gene in some Indian patients with chronic myeloid leukaemia. AB - The DNAs from 7 unselected Philadelphia chromosome-positive (Ph') chronic myeloid leukaemia (CML) patients were analysed for the distribution of breakpoints in the breakpoint cluster region (bcr) gene by Southern blot. Breakpoints were detected between the regions of exons '2' and '4' of the bcr gene in 5 out of 7 patients. In two patients, breakpoints occurred around exon 4 of the bcr gene. This study presents some information on the nature of breakpoint distribution in the bcr gene of Indian CML patients. PMID- 8707364 TI - Remission with carbimazole therapy & assessment of T4 suppression test as an index of relapse in patients with Graves' disease in India. AB - This study determined the relapse rate following the use of antithyroid drugs (ATD) in patients with Graves' disease and assessed T4 suppression test as a follow up index for predicting relapse after carbimazole treatment in 21 patients who had taken 9-12 months of ATD treatment continuously with good compliance. T4 suppression test was done before stopping ATD treatment. During one year of follow up after stopping ATD therapy, 12 (57%) patients relapsed and 9 (43%) remained in remission. Six of the 12 relapses occurred in the first 3 months of stopping ATD therapy. The response following the use of carbimazole therapy was comparable to that reported from iodine sufficient western countries and may be because of the salt iodination programme in our country. T4 suppression test was normal in 14 (66%) and abnormal in 7 (34%) patients. All the patients with abnormal T4 suppression test relapsed after stopping ATD. The overall accuracy of the T4 suppression test (76%) also favourably compared with reported values of other useful but less readily available markers such as thyrotropin releasing hormone (TRH) stimulation test and thyroid receptor antibodies. Thus, in our experience antithyroid drugs were able to induce long term remission in 43 per cent patients with Graves' disease and abnormal T4 suppression test can be used as a reliable parameter for predicting relapse. PMID- 8707365 TI - In vitro anti human immunodeficiency virus activity of mangrove plants. AB - Mangrove plant extracts were screened in vitro against human immunodeficiency virus (HIV) on MT-4 cells. HIV infected MT-4 cells were cultured for five days in the presence of the extract, at various concentrations. Anti-HIV activity was evaluated by tetrazolium-based colorimetric assay. Seven extracts were found effective. Five of the active extracts completely inhibited the virus adsorption to the cells. Most of the active plants are from the family Rhizophoraceae. PMID- 8707366 TI - Some pharmacological properties of Synclisia scabrida II. AB - The effect of the aqueous extract of S. scabrida on behaviour, and as an analgesic and antiulcer agent were studied. The extracts did not produce significant central nervous system action, or analgesia but had significant antiulcer activity against aspirin induced ulcer. The extract showed anticholinergic and antihistaminergic properties. PMID- 8707367 TI - Absence of wild poliovirus circulation among healthy children in a rural area with high oral poliovirus vaccination coverage. AB - Poliovirus circulation in a rural community was studied by a stool sample survey. No acute paralytic poliomyelitis case had been reported from the study area during the previous 5 months. Immunization coverage in age groups 7 to 12 months and 12 to 60 months was 95.8 and 94 per cent, respectively. Of the 257 children from whom stool samples were collected (about 6% of the child population), 161 (62.6%) were positive for virus isolation. Poliovirus was isolated from 60 (23.3%) children. All three poliovirus types were detected (41 type 1, 16 type 2 and 3 type 3). Intratypic differentiation tests classified these isolates as vaccine-like. Among the children excreting poliovirus, the proportion of those who did not receive polio vaccine within 30 days prior to the sample collection was 46.3, and 68.7 per cent for poliovirus type 1 and 2, respectively. It was concluded that these poliovirus excreting children were infected by the vaccine strains circulating in the environment. The survey showed that wild poliovirus was not detectable within five months after the last case of acute poliomyelitis. Displacement of the wild virus from the environment and circulation of vaccine virus was achieved by high vaccination coverage in this area. PMID- 8707368 TI - A comparison of sputum examination for acid fast bacilli by modified Schaeffer & Fulton stain, Ziehl-Neelsen stain & cold stain. AB - A comparative study of Ziehl-Neelsen stain, cold stain and modified Schaeffer and Fulton stain was carried out to evaluate the efficiency of modified Schaeffer and Fulton method in sputum examination for acid fast bacilli (AFB). Of 187 sputum samples studied, 67 (35.82%) were reported positive by Ziehl-Neelsen stain and cold stain method while 66 (35.29%) were reported positive by modified Schaeffer and Fulton method. In comparison with Ziehl-Neelsen staining, 98.58 per cent positivity was reported by modified Schaeffer and Fulton method. Modified Schaeffer and Fulton method is found to be simple, reliable, less expensive and as efficient as Ziehl-Neelsen stain and cold stain for demonstration of acid fast bacilli in sputum. PMID- 8707369 TI - Characterisation of invasive Haemophilus influenzae isolated in Nagpur, central India. AB - A total of 14 invasive H. influenzae were isolated from 12 patients by culturing 621 samples of body fluids over a span of one year. Twelve non-replicate isolates were analyzed for their drug susceptibility and serotype distribution. Eight (66.7%) isolates were multiply drug resistant with resistance to ampicillin, chloramphenicol and cotrimoxazole in different combinations. All isolates were susceptible to cefotaxime and erythromycin. Serotype b accounted for 10 (83.3%) isolates while one each were type a and non-typeable respectively. A continuous surveillance of drug susceptibility and serotype distribution is important to determine changes in the behaviour of the organism over time. PMID- 8707370 TI - Current status of Anopheles stephensi response to various insecticides in some areas of the Thar desert. AB - Investigations on the current response of A. stephensi. to six insecticides viz. DDT, dieldrin, malathion, fenitrothion, propoxur and permethrin, were carried out in 3 districts i.e. Barmer, Jodhpur and Pali, of the Thar desert. The species was found resistant to DDT and dieldrin, partially resistant to malathion and susceptible to fenitrothion, propoxur and permethrin. Dieldrin and malathion resistance has been detected for the first time in the Thar desert. Lethal concentrations (LC50 & LC95) of DDT and dieldrin and lethal exposure times (LT50 & LT95) of malathion, fenitrothion, propoxur and permethrin have been determined. In some areas, the differences in LC50 and LT50 values of tested insecticides, except fenitrothion, were found statistically significant. Chi-square and regression tests have revealed the homogeniety and linear trend respectively in the response of A. stephensi to insecticides. The findings of the study indicate that organochlorine compounds can be used alternately in the spray operations. PMID- 8707372 TI - Effect of calcium channel blockers on withdrawal syndrome of lorazepam in rats. AB - Effects of calcium channel blockers were investigated on withdrawal signs in lorazepam dependent rats. Physical dependence was produced by giving lorazepam admixed with the food in the following dose schedule: 10 x 4, 20 x 4, 40 x 4, 80 x 4 and 120 x 7 (mg/kg daily x days). Parameters such as body weight, food intake, spontaneous locomotor activity (SLA), body temperature, reaction time to pain, foot shock-aggression (FSA) and audiogenic seizures were observed during the period of administration of lorazepam and after its withdrawal. Calcium channel blockers viz. verapamil, nifedipine and nimodipine in different doses were administered orally twice daily in separate groups during the withdrawal period. The withdrawal signs observed in control group (without calcium channel blockers) were hyperkinesia, hyperthermia, hyper-aggression and audiogenic seizures. The administration of verapamil (5-20 mg/kg), nifedipine (1.75-7 mg/kg) and nimodipine (5-20 mg/kg) during the withdrawal period of lorazepam showed dose dependent significant blockade of all the withdrawal signs. Audiogenic seizures were completely blocked by 20 mg/kg dose of verapamil and nimodipine while nifedipine was partially effective. It may be concluded that calcium channel blockers exert protective effects on benzodiazepine withdrawal syndrome. PMID- 8707371 TI - Nutritional status of school age children of salt-workers in Rajasthan. AB - Nutritional status of 1044 school age children (6-18 yr) of salt workers in various salt manufacturing sites of Rajasthan, was assessed using anthropometry. Heights and weights of girls were observed to be better than in boys in the menarcheal age groups. Children of the present study showed reduced growth in comparison to National Centre for Health Statistics (NCHS), World Health Organisation (WHO), well-to-do Indian children and Rajasthan reference values of the same age groups. The prevalence of stunted with severe wasting was 4.5 per cent in the children of salt workers. Higher prevalence of severe wasting was observed in girls than the boys (P < 0.01). PMID- 8707373 TI - Response of mouse sarcoma-180 to cis-platin in combination with radiation & hyperthermia. AB - The in vivo response of a transplantable mouse tumour, sarcoma-180, to cis-platin (cDDP, 1.0, 2.5 or 5.0 mg/kg b.wt), local hyperthermia (HT, 42 degrees C, 30' or 60' and 43 degrees C, 30') and irradiation (RT, 10 Gy) was assessed on the basis of tumour cure (complete response, CR), volume doubling time (VDT) and regrowth delay (RD) as well as animal survival up to 120 days. Each agent was given as a single modality or in different combination regimens. A single injection of cDDP produced a dose dependent increase in all the parameters. Of all the single treatments, HT at 43 degrees C, 30' gave the maximum tumour cure. Combination of any of the two treatments resulted in a better response than all the single treatments. The chemosensitizing effect of heat was more pronounced than its radiosensitizing effect. Combination of all the three modalities, cDDP (2.5 mg/kg) + RT + HT (43 degrees C, 30') resulted in 100 per cent CR, without any local recurrence. This treatment also resulted in a significant increase in 120 day tumour free survival compared to all single modality treatments and bimodality treatments, except cDDP + 43 degrees C, 30'. This study indicates a potential advantage of the trimodality approach over single and bimodality treatments in the local control of solid tumours. PMID- 8707374 TI - Evaluation of an ELISA for the diagnosis of brucellosis. AB - Brucela serology using ELISA and standard agglutination test was performed on 23 patients with prolonged fever where the test was requested, on 26 randomly chosen patients with prolonged fever where it was not requested and on 17 controls. ELISA was positive in 39.1, 26.9 and 0 per cent respectively in these groups. Brucellosis may often be unsuspected because of its varied clinical manifestations and may be a more important cause of fever than previously considered. Our data reaffirm that ELISA is superior to the standard agglutination test for the diagnosis of brucellosis. PMID- 8707376 TI - Possible involvement of phospholipase D and protein kinase C in vascular growth induced by elevated glucose concentration. AB - Hyperglycemia is believed to be a major cause of diabetic vascular complications. To elucidate the effect of hyperglycemia on vascular response, we studied hyperproliferation, hypertrophy, and the natriuretic peptide response of vascular smooth muscle cells under high-glucose conditions. We observed that cells cultured in high glucose (22.2 mmol/L) showed hyper-proliferation and hypertrophy and that natriuretic peptide receptor responses were suppressed compared with cells cultured in normal glucose (5.6 mmol/L). We also examined phospholipase D and protein kinase C activities and found that in high-glucose conditions such activities are higher than in cells cultured in normal glucose. The activation of phospholipase D was not prevented by coincubation with 1 mumol/L protein kinase C(19-36), a specific protein kinase C inhibitor, but the activation of protein kinase C was. Protein kinase C(19-36) also markedly attenuated vascular hyperproliferation and hypertrophy as well as glucose-induced suppression of natriuretic peptide receptor response. These results show that hyperglycemia may be linked to vascular hyperproliferation, hypertrophy, and a suppressed natriuretic peptide receptor response, which are caused by increased phospholipase D and protein kinase C activities. PMID- 8707375 TI - Effects of angiotensin II infusion and inhibition of nitric oxide synthase on the rat aorta. AB - In previous studies, we showed that in vivo infusion of angiotensin II (Ang II) to adult rats induced vascular changes in gene expression, and this effect did not depend solely on blood pressure elevation. To determine whether nitric oxide can influence the effects of Ang II on the vessel wall, we administered to rats Ang II separately or in combination with the arginine analogue N omega-nitro-L arginine methyl ester, which inhibits nitric oxide synthase chronically when given in vivo. We measured changes in aortic medial thickness, the association of macrophages with the endothelial surface of the aorta, the presence of proliferating cell nuclear antigen in the intima and adventitia as an index of aortic cell cycle changes, and the expression of immunodetectable fibronectin as an index of changes in the extra-cellular matrix. After 18 days of nitric oxide inhibition, the major changes were increased medial thickness and a 3.5-fold increase in the number of adherent macrophages. Rats treated with two different doses of Ang II for 3 days had a fivefold and threefold increase in the number of proliferating cells from the intimal and adventitial regions, respectively. Combined treatment resulted in increased medial thickness, intimal and adventitial cell proliferation, and macrophage adherence. An increased and altered pattern of fibronectin distribution was found in all treatment groups. Losartan administration prevented the effects of Ang II but not of nitric oxide inhibition, whereas administration of L-arginine, prevented both intimal macrophage adherence and increased adventitial proliferation in rats given combined treatment. The data suggest that nitric oxide selectively influences macrophage association with the arterial wall, whereas Ang II and nitric oxide may have opposing effects on arterial cell proliferation. PMID- 8707377 TI - Thromboxane/prostaglandin endoperoxide-induced hypertrophy of rat vascular smooth muscle cells is signaled by protein kinase C-dependent increases in transforming growth factor-beta. AB - In the present study, we examined the effect of the thromboxane/prostaglandin endoperoxide analogue U46619 on proliferation and hypertrophy in cultured rat vascular smooth muscle cells and the roles of protein kinase C and transforming growth factor-beta (TGF-beta) in the mediation of the hypertrophic response to U46619. Since an increase in basic fibroblast growth factor (bFGF) was previously shown to mediate the hypertrophic response to U46619, we also assessed the relationship between bFGF and TGF-beta in the expression of U46619 actions. U46619 increased [35S]methionine incorporation into protein and protein content of vascular smooth muscle cells but had no effect on cell number. A role for TGF beta was supported by the following observations: (1) exogenous human TGF-beta 1 increased protein synthesis; (2) antibody to TGF-beta blocked both TGF-beta- and U46619-induced increases in protein content; (3) U46619 increased active and total TGF-beta bioactivities; and (4) the actions of U46619 on protein content and TGF-beta bioactivity were blocked by the thromboxane/prostaglandin endoperoxide receptor antagonist SQ 29,548. Previous observations had demonstrated a role for bFGF in the expression of U46619 actions on protein synthesis. Results of the present study suggest that TGF-beta and bFGF interact in mediating the protein synthetic response to U46619. First, the concentration of exogenous TGF-beta (10 pmol/L) alone required to produce a protein synthetic response equivalent to that induced by U46619 was much higher than the concentration of endogenous active TGF-beta that accumulated in the media in response to U46619 (0.7 pmol/L). Second, bFGF (20 ng/mL) increased total TGF-beta bioactivity and stimulated protein synthesis. The hyper-trophic response to bFGF was blocked by anti-TGF-beta. The ability of U46619 and bFGF to increase protein synthesis and protein content in vascular smooth muscle cells was associated with TGF-beta-induced suppression of proliferation, as evidenced by the ability of antibody to TGF-beta to enhance U46619- and bFGF-induced increases in [3H]thymidine incorporation into DNA. Results of the present study also supported a role for protein kinase C in the expression of U46619 and bFGF actions. U46619 increased protein kinase C activity in the particulate fraction of vascular smooth muscle cells. Moreover, the protein kinase C inhibitors GF109203X and staurosporine blocked U46619- and bFGF-induced increases in protein synthesis as well as active and total TGF-beta bioactivities. By contrast, the protein kinase C inhibitors did not prevent the increases in protein synthesis induced by exogenous TGF-beta. The results demonstrate that thromboxane/prostaglandin endoperoxide signals increased TGF-beta bioactivity via protein kinase C. Increases in both bFGF and TGF-beta are required for an optimal hypertrophic response to U46619. The hypertrophic response to TGF-beta occurs through a protein kinase C-independent pathway. PMID- 8707378 TI - Effect of long-term treatment with propionyl-L-carnitine on smooth muscle cell polyploidy in spontaneously hypertensive rats. AB - Experimental studies suggest that DNA content is increased in the smooth muscle cells of the arteries of hypertensive animals. It is unclear whether an increase in DNA content occurring in the smooth muscle cells of hypertensive rats represents a pressure-dependent effect. To evaluate the antihypertensive effect of long-term treatment with propionyl-L-carnitine and the possible morphological changes in thoracic smooth muscle cells correlated with this effect, we studied 4 month-old spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY) randomly divided into five groups. One group of SHR was treated with propionyl-L carnitine for 12 months; the other four groups of SHR and WKY received no treatment and were controls. We used static and flow cytometry to evaluate the polyploid cell content in thoracic aorta smooth muscle cells. Systolic pressure in untreated SHR progressively increased during the experiment. Treatment did not significantly influence pressure values in SHR. In WKY, blood pressure was significantly lower than that in treated and untreated age-matched SHR (2P < .02). The number of polyploid smooth muscle cells was significantly lower in the propionyl-L-carnitine-treated SHR than in the untreated rats (2P < .04) and similar to values for WKY. The reduction of polyploid cells in treated SHR was paralleled by a significant decrease of the aortic total DNA content, whereas no modifications occurred in smooth muscle cell mass. Long-term treatment with propionyl-L-carnitine may interfere with cellular mechanisms regulating the secondary responses involved in DNA synthesis. PMID- 8707379 TI - Effect of pregnancy on mechanisms of relaxation in human omental microvessels. AB - We assessed mechanisms of acetylcholine- and bradykinin-induced relaxations in human omental resistance vessels. Ring segments (approximately 200 microns normalized ID) were dissected from omental biopsies obtained from women at laparotomy (nonpregnant) or at cesarean delivery (pregnant) and were studied under isometric conditions in a Mulvany-Halpern myograph. All arginine vasopressin-preconstricted vessels relaxed in a strictly endothelium-dependent manner to acetylcholine and bradykinin; maximal relaxations were not decreased by either NG-nitro-L-arginine or indomethacin. By contrast, bradykinin failed to relax vessels that had been preconstricted with potassium gluconate. In the combined presence of NG-nitro-L-arginine and indomethacin, addition of charybdotoxin, a selective antagonist of some calcium-sensitive potassium channels, did not inhibit maximal bradykinin-induced relaxation. By contrast, addition of 10 mmol/L tetraethylammonium chloride abolished relaxation in vessels from nonpregnant women but not in vessels from gravidas. We conclude that bradykinin relaxes these human resistance arteries in an endothelium-dependent but predominantly nitric oxide- and prostanoid-independent manner; relaxation likely depends on the action of an endothelium-derived hyperpolarizing vasodilator. Furthermore, in striking contrast to mechanistic insights from animal studies, human pregnancy appears to augment a mechanism of endothelium dependent relaxation in these vessels that is insensitive to the inhibitors noted above. Whether a similar novel vasodilator mechanism in vivo contributes to the physiological vasodilation that characterizes human gestation or whether failure of such a mechanism might lead to preeclampsia remains the subject of future study. PMID- 8707380 TI - Comparison of effect of endothelin antagonism and angiotensin-converting enzyme inhibition on blood pressure and vascular structure in spontaneously hypertensive rats treated with N omega-nitro-L-arginine methyl ester. Correlation with topography of vascular endothelin-1 gene expression. AB - Inhibition of nitric oxide synthase by L-arginine analogues such as N omega-nitro L-arginine methyl ester (L-NAME) in spontaneously hypertensive rats (SHR) is associated with malignant hypertension and enhanced expression of the endothelin 1 gene in some blood vessels. In this study, SHR treated chronically with L-NAME (SHR-L-NAME) were given the angiotensin I-converting enzyme inhibitor cilazapril or the endothelin-A/endothelin-B receptor antagonist bosentan for 3 weeks. Systolic pressure was lowered slightly by cilazapril (213 +/- 2 versus 229 +/- 2 mm Hg in untreated SHR-L-NAME, P < .01) but was not significantly lowered by bosentan (223 +/- 2 mm Hg). Hypertrophy of aorta and small arteries (coronary, renal, mesenteric, and femoral) was decreased by cilazapril treatment and unaffected by bosentan. Expression of the endothelin-1 gene was evaluated in SHR L-NAME by in situ hybridization histochemistry, which showed that endothelin-1 expression was enhanced in the endothelium of aorta but not in small mesenteric arteries in these rats. The absence of enhancement of endothelin-1 gene expression in small arteries may account for the absence of increased severity of hypertrophy of small vessels in SHR-L-NAME and may be a mechanism whereby L-NAME inhibits cardiovascular growth. These results suggest that in the absence of enhanced small-artery endothelin-1 expression, endothelin antagonism does not lower blood pressure. The blood pressure-lowering effect of angiotensin I converting enzyme inhibition suggests a role for the renin-angiotensin system in the malignant form of hypertension that develops in SHR treated with L-NAME. PMID- 8707381 TI - Characterization of the renal phenotype of transgenic rats expressing the human endothelin-2 gene. AB - We have previously established a transgenic rat model termed TGR(hET-2)37 overexpressing the human endothelin-2 (ET-2) gene with high renal transgene expression. This renal overexpression is of pathophysiological interest because a long-term activated paracrine renal endothelin system has been implicated in chronic renal failure due to progressive glomerular injury. Therefore, our aim in the present study was to analyze renal transgene expression in detail and address the question of whether transgene expression causes phenotypic and functional changes in the kidney. We used reverse transcription-polymerase chain reaction and in situ hybridization techniques for transgene expression analysis. Tissue ET 2 concentrations were measured with a specific radioimmunoassay. For histological evaluation of renal tissue, all samples were subjected to hematoxylin-eosin and periodic acid-Schiff staining. Renal tissue ET-2 concentrations were significantly increased in TGR(hET-2)37 rats. Using in situ hybridization, we found that the human ET-2 gene was almost exclusively expressed within the glomeruli. The glomerular transgene expression resulted in a significantly increased glomerular injury score and likewise in a significantly increased protein excretion, whereas glomerular filtration rate was not altered. Blood pressure was similar in TGR(hET-2)37 rats and age-matched controls, suggesting that the local changes in the kidney were correlated with paracrine endothelin actions. In conclusion, our study revealed that the major renal expression site of the human ET-2 transgene in TGR(hET-2)37 rats was within the glomeruli and caused the development of glomerulo-sclerosis with significantly increased protein excretion that is independent of blood pressure. We suggest that TGR(hET 2)37 rats are a new monogenetic animal model for study of the paracrine renal endothelin system and its involvement in renal pathophysiology. PMID- 8707382 TI - Role of nitric oxide, adenosine, and ATP-sensitive potassium channels in insulin induced vasodilation. AB - The resistance of various tissues to the vasodilator and metabolic effects of insulin may be an important risk factor in the genesis of hypertension observed in several pathological states. Because of this, it is important to understand the mechanisms by which insulin causes vasodilation. Because insulin is known to raise metabolism, one mechanism by which insulin causes vasodilation could be through metabolic vasodilation. Recently, however, it has been suggested that the insulin-induced vasodilation is mediated by the release of endothelium-derived nitric oxide. Using a model of muscle microcirculation (hamster cremaster), we examined the interactions between insulin, nitric oxide, and tissue metabolism to understand the potential mechanisms by which insulin causes vasodilation. Topical application of insulin (200 microU/mL) to the cremaster resulted in significant increases in arteriolar diameter. Second-order arteriolar diameter increased from 69.6 +/- 6 to 79.8 +/- 5 microns and fourth-order arteriolar diameter from 11.3 +/- 1 to 15.1 +/- 2 microns (n = 8). During nitric oxide synthase inhibition, topical application of insulin caused significant vasodilation in both second- and fourth-order arterioles. In contrast, both adenosine receptor antagonism and blockade of ATP-sensitive potassium channels prevented insulin-induced increases in arteriolar diameter. Our findings suggest a role for increased tissue metabolism, particularly the metabolite adenosine, in mediating insulin-induced vasodilation. PMID- 8707385 TI - Attenuation of hypertension by insulin-sensitizing agents. PMID- 8707384 TI - Elevated lymphocyte cytosolic calcium in a subgroup of essential hypertensive subjects. AB - Abnormalities of intracellular calcium homeostasis and sodium-proton exchange have been implicated in the pathophysiology of essential hypertension. To further define the nature of cytosolic calcium abnormalities and whether they relate to increased sodium-proton exchange in hypertension, we have studied peripheral lymphocytes from normotensive and hypertensive subjects. Lymphocyte cytosolic calcium was significantly increased (P < .01) in hypertensive compared with normotensive subjects while consuming a high salt diet. Using maximum likelihood analysis, we found that cytosolic calcium levels in our study population were not normally distributed and observed three modes (P < .02). The means of the first mode and the two upper modes were separated (+/-2 SD) at a cytosolic calcium level of 120 nmol/L. We conducted further analysis in the subgroups with cytosolic calcium levels > 120 nmol/L or < 120 nmol/L. The majority of the normotensive subjects (86%) and half of the hypertensive subjects (52%) had levels < 120 nmol/L. Clinical characteristics of the two subgroups did not differ. Subjects with levels < 120 nmol/L had a rise in cytosolic calcium when changed to a low salt diet; those with levels > 120 nmol/L did not show a change in cytosolic calcium but their blood pressure fell significantly with salt restriction. Hypertensive subjects also had increased sodium-proton exchange activity compared with normotensive subjects when both groups were studied in a high salt balance. A positive correlation between sodium-proton exchange and cytosolic calcium was observed in subjects with levels < 120 nmol/L. There was insufficient power to draw conclusions on this relationship in subjects with levels > 120 nmol/L. Thus, many hypertensive subjects have increased cytosolic calcium, but this abnormality is not associated with sodium-proton exchange activity in all individuals. The salt-induced change in cytosolic calcium in subjects with levels < 120 nmol/L and its link to sodium-proton exchange suggest regulation by factors involved in salt-volume homeostasis. Individuals with cytosolic calcium > 120 nmol/L, most of whom were hypertensive, may have abnormalities in this regulation, contributing to hypertension. PMID- 8707383 TI - Effects of insulin on calcium metabolism and platelet aggregation. AB - The influence of insulin on platelets in vitro has not been exhaustively investigated. To clarify whether insulin affects Ca2+ metabolism in platelets directly or through alteration of other systems regulating intracellular Ca2+ homeostasis, we examined the effect of insulin both alone and in combination with prostaglandin E1 on platelet aggregation and Ca2+ metabolism. Incubation of rat platelets with insulin reduced thrombin-induced Ca2+ influx but did not change thrombin-evoked release of Ca2+ from internal stores or the size of internal Ca2+ stores. The interactive effects of insulin with prostaglandin E1 were only additive, and insulin did not augment the effects of prostaglandin E1 on platelet Ca2+ metabolism. In contrast, insulin did not inhibit thrombin-induced platelet aggregation but did augment inhibition of platelet aggregation by prostaglandin E1. Our results suggest that insulin inhibits platelet function by both prostaglandin E1-dependent and -independent mechanisms. PMID- 8707386 TI - Angiotensin II increases norepinephrine turnover in the anteroventral third ventricle of spontaneously hypertensive rats. AB - We evaluated the effect of angiotensin II (Ang II) administered by intracerebroventricular injection on norepinephrine turnover in the anteroventral third ventricle in adult spontaneously hypertensive rats (SHR, n = 35) and age matched Wistar-Kyoto rats (WKY, n = 38). Ang II (100 ng) or saline (vehicle control) was administered into the cerebral ventricle 30 minutes after injection of alpha-methyl-p-tyrosine (250 mg/kg IP). Norepinephrine turnover was assessed by evaluation of the norepinephrine concentration before and 1 hour after such administration. The pressor response to Ang II administration was significantly greater in SHR than in WKY (+43 +/- 3 versus +23 +/- 2 mm Hg, P < .01). Baseline norepinephrine turnover (response to saline) was reduced in the ventral median preoptic nucleus of SHR. Ang II significantly increased norepinephrine turnover in the organum vasculosum lamina terminalis and ventral median preoptic nucleus of SHR (organum vasculosum lamina terminalis: 40 +/- 5% by Ang II versus 18 +/- 6% by saline, P < .05; ventral median preoptic nucleus: 32 +/- 3% by Ang II versus 21 +/- 2% by saline, P < .05) but not of WKY (37 +/- 5% versus 29 +/- 5%, P = NS, and 30 +/- 2% versus 32 +/- 3%, P = NS, respectively). Thus, norepinephrine turnover in the anteroventral third ventricle region induced by intracerebroventricular administration of Ang II was increased in SHR. This effect may contribute to the enhanced pressor response to central Ang II seen in this model. PMID- 8707387 TI - Trial of stress reduction for hypertension in older African Americans. II. Sex and risk subgroup analysis. AB - Our objective was to test the short-term efficacy and feasibility of two stress reduction approaches for the treatment of hypertension in older African Americans, focusing on subgroup analysis by sex and by high and low risk on six measures of hypertension risk: psychosocial stress, obesity, alcohol use, physical inactivity, dietary sodium-potassium ratio, and a composite measure. The study involved a follow-up subgroup analysis of a 3-month randomized, controlled, single-blind trial conducted in a primary care, inner-city health center. Subjects were 127 African American men and women, aged 55 to 85 years, with diastolic pressure of 90 to 104 mm Hg and systolic pressure less than or equal to 179 mm Hg. Of these, 16 did not complete follow-up blood pressure measurements. Mental and physical stress-reduction approaches-the Transcendental Meditation technique and progressive muscle relaxation, respectively-were compared with a life-style modification education control and with each other. Both systolic and diastolic pressures changed from baseline to follow-up for both sexes and for high and low risk level (defined by median split) on the six measures of hypertension risk. Compared with education control subjects, women practicing the Transcendental Meditation technique showed adjusted declines in systolic (10.4 mm Hg, P < .01) and diastolic (5.9 mm Hg, P < .01) pressures. Men in this treatment group also declined in both systolic (12.7 mm Hg, P < .01) and diastolic (8.1 mm Hg, P < .001) pressures compared with control subjects. Women practicing muscle relaxation did not show a significant decrease compared with control subjects, and men declined significantly in diastolic pressure only (6.2 mm Hg, P < .01). For the measure of psychosocial stress, both the high and low risk subgroups using the Transcendental Meditation technique declined in systolic (high risk, P = .0003; low, P = .06) and diastolic (high risk, P = .001; low, P = .008) pressures compared with control subjects, whereas for muscle relaxation, blood pressure dropped significantly only in the high risk subgroup and only for systolic pressure (P = .03) compared with control subjects. For each of the other five risk measures, Transcendental Meditation subjects in both the high and low risk groups declined significantly in systolic and diastolic pressures compared with control subjects. Effects of stress reduction on blood pressure were found to generalize to both sexes and diverse risk factor subgroups and were significantly greater in the Transcendental Meditation treatment group. These effects (along with high compliance) even in individuals with multiple risk factors for hypertension clearly warrant longer-term investigation in this and other populations. PMID- 8707388 TI - Noninvasive assessment of baroreflex control in borderline hypertension. Comparison with the phenylephrine method. AB - In this study, we examined the sensitivity of two recently developed noninvasive baroreflex measurement techniques to assess baroreflex control in hypertension. We assessed baroreflex sensitivity noninvasively from covariations of systolic pressure and RR interval using spectral analysis and sequence detection. The noninvasive estimates of baroreflex control were compared with estimates derived from phenylephrine-induced increases in systolic pressure and RR interval in normotensive subjects (n = 27) and borderline hypertensive subjects (n = 15). Baroreflex sensitivity was significantly reduced in the borderline hypertensive group relative to the normotensive group when assessed with the use of either the noninvasive or invasive methods to index baroreflex control. In addition, estimates obtained from the noninvasive methods were significantly correlated with baroreflex sensitivity assessed with the phenylephrine method (spectral: r = .48, P < .001; sequence: r = .50, P < .001). These findings suggest that spectral analysis and the sequence method provide viable alternatives to the pharmacological approach for estimation of baroreflex sensitivity in hypertension. PMID- 8707389 TI - Dietary sodium intake modulates pituitary proopiomelanocortin mRNA abundance. AB - The pituitary prohormone proopiomelanocortin gives rise to melanocortins of alpha, beta, and gamma primary structure in addition to corticotropin. Melanocortins have a variety of actions in mammals, and each is natriuretic. In particular, gamma-melanocyte-stimulating hormone has been shown to mediate reflex natriuresis after acute unilateral nephrectomy. We examined whether this peptide could play a role in longer term adjustments in sodium balance by measuring plasma gamma-melanocyte-stimulating hormone and corticotropin concentrations, as well as pituitary proopiomelanocortin mRNA abundance, in Sprague-Dawley rats ingesting either a low (0.07% NaCl) or high (7.5% NaCl) sodium diet. One week after the high sodium diet, plasma gamma-melanocyte-stimulating hormone concentration was double the value seen in rats on the low sodium diet (158 +/- 5 [SE] versus 76 +/- 9 fmol/mL, P < .001), a change that was accompanied by a fivefold increase in plasma atrial natriuretic peptide concentration but no change in plasma corticotropin. Whole pituitary proopiomelanocortin mRNA abundance, measured with a probe to exon 3 of the rat proopiomelanocortin gene, was significantly increased after 1 week of the high sodium diet compared with the low sodium diet and increased further at 2 and 3 weeks. This increase occurred primarily in the neurointermediate lobe as demonstrated by in situ hybridization; the content of gamma-melanocyte-stimulating hormone immunoreactivity was also increased in this lobe, but not the anterior lobe, after 1 week of the high sodium diet. These results demonstrate that high dietary sodium intake increases neurointermediate lobe proopiomelanocortin mRNA abundance compared with a very low sodium diet and also suggest that proopiomelanocortin is preferentially processed into gamma-melanocyte-stimulating hormone rather than corticotropin. These observations consequently raise the possibility of a role for this peptide hormone system in the adjustments to a high salt diet. PMID- 8707390 TI - Variability in salt sensitivity classifications in black male versus female adolescents. AB - Salt sensitivity (changes in blood pressure in response to alterations in salt intake) may be a risk factor for hypertension. In the present study, we examined the prevalence of salt sensitivity based on two different classifications in healthy black male and female adolescents (aged 13 to 16 years). A total of 135 black adolescents participated in a 50 mmol/24 h low sodium diet for 5 days and a 150 mmol/24 h NaCl supplement for 10 days. Dietary compliance was defined as sodium excretion less than or equal to 50 mmol/24 h for the low sodium diet and greater than or equal to 165 mmol/24h for the high NaCl supplement. Salt sensitivity was defined by two classifications: (1) as a decrease in mean blood pressure greater than or equal to 5 mm Hg from baseline to the low sodium diet, and (2) as an increase in mean blood pressure greater than or equal to 5 mm Hg from the low sodium diet to the high NaCl supplement. With classification 1, 14% of boys were identified as salt sensitive compared with 22% of girls. With classification 2, however, 31% of boys were identified as salt sensitive compared with 18% of girls. Analyses based on changes in systolic pressure demonstrated similar findings across sex, although overall classifications based on systolic pressure yielded a greater percentage of salt-sensitive subjects. These sex differences in classification patterns were not due to differences in other important variables, such as changes in sodium excretion, potassium excretion, or Quetelet index. These results suggest that the prevalence of salt sensitivity differs by sex depending on the type of protocol used for the classification of salt sensitivity in a black pediatric population. PMID- 8707391 TI - Hypertension in adult American Indians. The Strong Heart Study. AB - Hypertension is a primary risk factor for cardiovascular disease in the United States. Although cardiovascular disease is the leading cause of death among American Indians, the prevalence of hypertension, its awareness and control, and its association with other cardiovascular disease risk factors and physiological variables have not been well studied in this population. The Strong Heart Study is a longitudinal study of cardiovascular disease and its risk factors in American Indians. Participants (2703 women and 1846 men) were members of 13 tribes in central Arizona, southwestern Oklahoma, and regions of South and North Dakota. At least 1500 individuals between 45 and 74 years of age participated from each center in a baseline clinical examination conducted between July 1989 and January 1992. The examination consisted of a personal interview and physical examination that included an oral glucose tolerance test and three consecutive blood pressure measurements. This study reports data from the baseline examination on the prevalence of hypertension and correlates of blood pressure. Results indicated that despite the high frequency of diabetes and obesity, prevalence rates of hypertension in Arizona and Oklahoma were similar to those in the US population in the Third National Health and Nutrition Examination Survey (NHANES III), and rates among South/North Dakota participants were significantly lower (P < .0001). Blood pressure was higher in individuals with diabetes (P < .0001) and was significantly correlated with age (P < .0001) and albuminuria (P < .0001) but only weakly related to obesity. There was no independent relation between blood pressure and insulin. Blood pressure seems to be less affected by obesity and hyperinsulinemia in American Indians compared with other populations. Nevertheless, hypertension should be aggressively treated and controlled in American Indians because it is a known precursor to morbidity and mortality associated with diabetes and cardiovascular disease. PMID- 8707392 TI - Angiotensin II increases left ventricular mass without affecting myosin isoform mRNAs. AB - We studied the effect of chronic (7 days) angiotensin II (Ang II) infusion in nonpressor and pressor doses on cardiovascular mass and expression of alpha- and beta-myosin heavy chain genes in the left ventricle in normotensive Wistar rats. An increased left ventricular mass was observed in rats receiving non-pressor and pressor doses of Ang II, but only high doses increased arterial pressure. Normalization of arterial pressure during Ang II infusion by losartan, a specific Ang II receptor antagonist, or hydralazine had different effects on left ventricular mass. Losartan prevented the increased left ventricular mass, and hydralazine did not affect left ventricular mass. Northern blot analysis showed that the switch in left ventricular myosin isoform mRNA from the adult to the fetal pattern occurred only in rats given the pressor Ang II dose. Both losartan and hydralazine, in parallel with the normalization of arterial pressure, prevented this myosin isoform switch. Thus, these data suggest that the Ang II induced increase in left ventricular mass was not dependent on pressure overload, but the switch in myosin isoform mRNA from the adult to the fetal pattern was dependent on pressure overload. PMID- 8707393 TI - Advanced hypertensive heart disease in spontaneously hypertensive rats. Lisinopril-mediated regression of myocardial fibrosis. AB - Left ventricular hypertrophy (LVH) in spontaneously hypertensive rats (SHR) is accompanied by a structural remodeling of the myocardium that includes myocyte hypertrophy and interstitial and perivascular fibrosis of intramyocardial coronary arteries. The structural abnormalities related to fibrous tissue accumulation lead to increased myocardial diastolic stiffness and ultimately impaired systolic function of the left ventricle. It has been shown in 14-week old SHR with early hypertensive heart disease that myocardial fibrosis could be reversed and myocardial diastolic stiffness normalized by 12-week treatment with the angiotensin-converting enzyme inhibitor lisinopril. Whether such functional defects of the myocardium, based on adverse structural changes, are also reversible in advanced hypertensive heart disease has been questioned. Therefore, we treated 78-week-old male SHR that had chronic hypertension and advanced LVH with severe myocardial fibrosis and age- and sex-matched normotensive Wistar Kyoto rats (WKY) with 20 mg/kg per day oral lisinopril for 8 months. Compared with untreated SHR or WKY, we found the following: (1) Systolic arterial pressure was normalized (P < .025) and LVH completely reversed (P < .025) in SHR, with no significant reduction in systolic arterial pressure or left ventricular mass in WKY; (2) morphometrically determined myocardial fibrosis in SHR was significantly reversed (P < .025) and associated with improved diastolic stiffness (P < .05), which was measured in the isolated heart by calculation of the stiffness constant of the myocardium; no significant changes occurred in WKY; (3) reversal of myocardial fibrosis was accompanied by an increase (P < .025) in myocardial matrix metalloproteinase 1 activity determined by degradation of [14C]collagen with myocardial tissue extracts after trypsin activation of myocardial promatrix metalloproteinase 1; matrix metalloproteinase 1 activity remained unchanged in WKY treated with lisinopril; and (4) systolic dysfunction, measured by a significantly (P < .025) diminished slope of the systolic stress-strain relation under isovolumic conditions of the left ventricle, was found in 110-week-old SHR, and it could be prevented by lisinopril treatment. Thus, long-term angiotensin converting enzyme inhibition with lisinopril normalized arterial pressure and LVH, reversed myocardial fibrosis, and improved abnormal myocardial diastolic stiffness in advanced hypertensive heart disease in SHR. In addition, systolic dysfunction of the left ventricle could be prevented. The fibrolytic response to lisinopril was at least partly due to enhanced collagen degradation by activation of tissue matrix metalloproteinase 1. PMID- 8707394 TI - Influence of obesity on left ventricular midwall mechanics in arterial hypertension. AB - The evaluation of the effect of obesity on left ventricular systolic performance may differ in relation to the method used to measure left ventricular function and to the type of study population. Whether obesity worsens left ventricular midwall mechanics in arterial hypertension has never been investigated. Accordingly, we assessed echocardiographic left ventricular midwall shortening circumferential end-systolic stress relations in 156 normotensive and normal weight (reference) adults, 94 normotensive and overweight (1985 National Institutes of Health partition values) to obese (body mass index > 30 kg/m2) adults, 263 hypertensive and normal-weight adults, and 224 hypertensive and overweight-to-obese adults. There was an inverse relation of midwall shortening to circumferential end-systolic stress in all groups (all P < .005). Left ventricular performance as a ratio of observed to predicted midwall shortening fell below the fifth percentile in 4 of 94 (4%) of overweight-to-obese normotensive individuals. Eighty-eight of 487 hypertensive subjects (18.1%) exhibited depressed midwall shortening as a percentage of the value predicted from wall stress, with no difference between normal-weight (50 of 263 [19%]) and overweight (38 of 224 [17%]) subjects. Sixty-one normotensive and 131 hypertensive subjects were frankly obese. After adjustment for sex and age, midwall shortening, as either absolute values or a percentage of predicted, was not statistically different among obese, overweight, and normal-weight subjects in both normotensive and hypertensive groups. For each quartile of observed-to predicted midwall shortening ratio, obese subjects had greater left ventricular end-diastolic volume than normal-weight subjects among both normotensive and, more evidently, hypertensive subjects. A predicted midwall shortening was generated from both wall stress and left ventricular volume with the use of multiple regression analysis. High body mass index, mean blood pressure, aging, and male sex independently predicted low afterload and left ventricular volume independent midwall left ventricular performance (multiple R = .31, P < .0001). Thus, (1) midwall left ventricular systolic performance in asymptomatic overweight or frankly obese individuals is comparable to that in normal-weight individuals in both the presence and absence of arterial hypertension; (2) however, maintenance of normal life ventricular performance in obese individuals is associated with the use of Starling reserve; and (3) this compensatory mechanism is especially evident when arterial hypertension and obesity coexist. PMID- 8707395 TI - Association between persistent pressure overload and ventricular arrhythmias in essential hypertension. AB - Hypertension is a risk factor for sudden cardiac death, and some data indicate that frequent and complex ventricular arrhythmias may be additional risk markers in hypertensive individuals. We investigated the relation between ventricular arrhythmias and the persistence of increased blood pressure levels over 24 hours in subjects with essential hypertension. We studied 126 never-treated subjects with essential hypertension (83 men) who underwent 24-hour electrocardiographic monitoring, 24-hour ambulatory blood pressure monitoring, and echocardiography. Premature ventricular beats were detected in 71% of the subjects. Compared with subjects in Lown class 0-1, subjects with frequent or complex ventricular arrhythmias (Lown class > or = 2) were older (54 versus 45 years) and had a longer duration of hypertension (5.4 versus 2.8 years), a greater left ventricular mass (147 versus 127 g.m-2), and a blunted nocturnal reduction in ambulatory blood pressure (7%/12% versus 12%/16%). The number of premature ventricular beats over 24 hours was associated with age (r = .25), left ventricular mass (r = .24), and pulse pressure (r = .18) and inversely associated with the present reduction in blood pressure from day to night (r = -.29 for systolic and -.25 for diastolic pressures). In a multiple logistic regression analysis, frequent or complex ventricular arrhythmias (Lown class > or = 2) were predicted by an age > or = 60 years (odds ratio, 10.4 95% confidence interval, 2.4-44.8), left ventricular hypertrophy at echocardiography (odds ratio, 4.2; 95% confidence interval, 1.5-11.6), and a < 10% reduction in blood pressure from day to night ("nondipping" pattern; odds ratio, 2.9;95% confidence interval, 1.2 7.0). We conclude that in addition to the strong effect of age and left ventricular hypertrophy at echocardiography, the persistence of high blood pressure levels over the 24 hours ("nondipping" pattern) is an independent predictor of the frequency and complexity of ventricular arrhythmias in never treated subjects with essential hypertension. PMID- 8707396 TI - Regulation of human renin mRNA expression and protein release in transgenic mice. AB - The renin-angiotensin system plays a major role in the regulation of blood pressure and electrolyte homeostasis in mammals. In this study, we subjected transgenic mice containing a human renin genomic construct to a variety of pharmacological and physiological manipulations to test whether expression of the human renin gene and release of active human renin in appropriately regulated in this model. These manipulations were designed to test major regulators of renin release, including angiotensin II, the macula densa, renal perfusion pressure, and beta-adrenergic receptors. We used human plasma renin concentration and human renal renin mRNA levels to document the response of the transgene to these stimuli. Human plasma renin concentration increased in response to both angiotensin-converting enzyme inhibition with captopril and isoproterenol and decreased after a high salt diet. A low salt or sodium-deficient diet did not stimulate renin release. Human renin mRNA levels in kidney increased after captopril but were unchanged in the other experimental groups. We also measured the levels of human renin mRNA in double transgenic mice containing the same human renin gene in addition to the human angiotensinogen gene. These mice are chronically hypertensive and have increased circulating levels of angiotensin II. Human renin mRNA levels in the kidney were paradoxically elevated compared with their single transgenic normotensive counterparts. These transgenic mice provide a model for examination of human renin regulation and may help elucidate the molecular mechanisms that regulate the gene in response to physiological cues. PMID- 8707397 TI - Neural nitric oxide synthase in the renal medulla and blood pressure regulation. AB - We studied the effect of selective inhibition of the neural isoform of nitric oxide synthase in the rat renal medulla in conscious Sprague-Dawley rats. Continuous renal medullar interstitial infusion of an antisense oligonucleotide complementary to the initiation region of the mRNA for neural nitric oxide synthase increased blood pressure 14 +/- 1 mm Hg in rats maintained on a high sodium intake. Medullary interstitial infusion of saline vehicle or a scrambled oligonucleotide probe failed to alter blood pressure in separate groups of high salt control rats. Renal medullary interstitial infusion of the antisense oligonucleotide significantly decreased the level of neural nitric oxide synthase in the renal medulla by 53 +/- 8% and decreased total renal medullary nitric oxide synthase activity by 28 +/- 8%. No alterations were detected in the levels of inducible nitric oxide synthase or beta-actin in the antisense oligonucleotide infused rats. To confirm the antisense oligonucleotide data, we administered a mechanistically different inhibitor of neural nitric oxide synthase, 7 nitroindazole, to an additional group of rats maintained on a high salt diet. Direct renal medullary interstitial infusion of this selective enzyme inhibitor significantly increased mean arterial pressure (15 +/- 6 mm Hg) and decreased total renal medullary nitric oxide synthase activity by 37 +/- 12% in rats on a high sodium diet. The present experiments demonstrate a role for the neural isoform of nitric oxide synthase in the long-term control of blood pressure in the presence of a high salt diet. PMID- 8707398 TI - Cyclosporin A impairs the nocturnal blood pressure fall in renal transplant recipients. AB - In renal transplant recipients, hypertension and a diminished nocturnal blood pressure fall are frequently found. To investigate whether this diminished nocturnal blood pressure fall is related to the use of cyclosporin A or to other factors, such as the use of glucocorticoids, we measured 24-hour ambulatory blood pressure in 18 renal transplant recipients both before and 16 weeks after conversion from cyclosporin A to azathioprine. Renal blood flow and glomerular filtration rate were estimated from 131I-hippurate and 125I-iothalamate clearances, respectively, and plasma concentrations of renin, atrial natriuretic peptide, norepinephrine, prostaglandin E2, and thromboxane B2 were determined. During cyclosporin A treatment, mean 24-hour blood pressure was 117 +/- 3 mm Hg, and the nocturnal fall in blood pressure was 4 +/- 9 mm Hg. A nondipping diurnal blood pressure pattern was present in 13 patients. After conversion to azathioprine, mean 24-hour blood pressure decreased to 109 +/- 3 mm Hg (P < .001), the nocturnal fall increased to 9 +/- 6 mm Hg, and the number of patients with a nondipping diurnal blood pressure pattern decreased to 9. The nocturnal fall in heart rate (17 +/- 10 beats per minute) during cyclosporin A did not change after conversion. Body weight and plasma concentrations of norepinephrine and renin did not change. Plasma concentrations of prostaglandin E2 and thromboxane B2 decreased after conversion, as did plasma atrial natriuretic peptide. Renal blood flow and glomerular filtration rate increased after conversion. In conclusion, cyclosporin A appears to be involved in the disturbance of the circadian blood pressure rhythm in renal transplant recipients. Although the precise mechanism is unclear. the elevated plasma atrial natriuretic peptide and slightly suppressed plasma renin concentrations suggest that intravascular volume expansion may contribute to the observed hemodynamic alterations. PMID- 8707399 TI - Inhibition of aldosterone production by adrenomedullin, a hypotensive peptide, in the rat. AB - Recently, we conducted in vitro studies and reported that adrenomedullin, a novel hypotensive peptide, inhibits aldosterone secretion by dispersed rat adrenal zona glomerulosa cells. To assess the physiological role of this inhibitory effect, we investigated the effect of adrenomedullin on aldosterone production in vivo. Male Sprague-Dawley rats were fed a normal sodium diet before the experiments. To begin the experimental procedure, we stimulated aldosterone production with a sodium-deficient diet or bilateral nephrectomy. After 3 days of sodium depletion or immediately after nephrectomy, we injected synthetic human adrenomedullin (2.5 nmol/kg SC) and repeated the injection three times at 6-hour intervals. Two hours after the last injection, the rats were decapitated and adrenal capsular tissue was collected. Adrenomedullin had no effect on plasma and adrenal aldosterone concentrations in the rats fed a normal sodium diet. Rats fed a sodium-deficient diet had significantly increased aldosterone concentrations in both plasma (4770.1 +/- 364.3 pmol/L) and adrenal gland (57.34 +/- 3.27 pmol per adrenal). Subsequently, injection of adrenomedullin significantly inhibited increases in concentrations (plasma, 2648.9 +/- 313.2 pmol/L; adrenal, 44.28 +/- 4.94 pmol per adrenal). In nephrectomized rats, increased aldosterone concentrations in plasma and adrenal gland were also significantly inhibited by adrenomedullin. In the second part of the study, plasma renin concentration, adrenal renin activity, plasma corticosterone concentration, serum potassium concentration, and plasma immunoreactive adrenomedullin concentration were examined for adrenomedullin effects. The first four were unaffected, and the last, plasma immunoreactive adrenomedullin, was elevated 15% to 30%. These in vivo results, together with our in vitro data, suggest that adrenomedullin may indeed play a physiological role in the control of blood pressure and electrolyte balance. PMID- 8707400 TI - An E-box motif conveys inhibitory activity on the atrial natriuretic peptide gene. AB - Atrial natriuretic peptide (ANP) is a potent diuretic, natriuretic, and vasorelaxant hormone that is expressed early in ventricular hypertrophy. Expression of human ANP is controlled by a series of regulatory elements located in the 5' flanking sequence of its gene. We generated a series of 5' deletion mutations extending from -2600 to -1150 relative to the transcription start site and linked them to a chloramphenicol acetyltransferase reporter gene. Using transient transfection analysis, we have identified a negative regulatory element between -1206 and -1152 relative to the start site. Each of a series of 5' deletion mutants, when introduced into fibroblast cultures, expressed the reporter function at a level that was significantly less (< 20%) than that seen with the -1152 reporter construct, whereas comparably transfected atrial cardiocytes demonstrated no change in reporter activity, implying that the repressor function is specific to cell type. The critical region (from -1206 to 1152) associates with a soluble protein present in cardiac fibroblast extracts in a sequence-specific fashion. Deoxyribonuclease I footprint analysis demonstrated the presence of several protected regions, including one that overlies an E-box motif (CAACTG), an element that in other systems has been implicated in promoting differentiation in the myocyte lineage. Site-directed mutagenesis of the E-box motif suppressed both the protein-binding and inhibitory activities of the 54-bp fragment. In summary, we have found a region in the 5' flanking sequence of the human ANP gene that represses transcriptional activity in nonmyocardial cells. This element may play an important role in the restriction of ANP gene expression to cardiac myocytes. PMID- 8707401 TI - Overexpression and mutations of p53 in metastatic malignant melanomas. AB - Alterations of the p53 tumor suppressor gene are the most frequent genetic abnormalities in human malignancies, but the role of p53 in the etiology of malignant melanomas is unclear. Fifty unselected malignant melanomas were analyzed for p53 overexpression by immunohistochemistry using 3 monoclonal antibodies (MAbs). Fifteen tumors (29.4%) showed positive staining with at least 2 different antibodies. In the first 20 consecutive tumors exons 5-9 and adjacent splice sites of the p53 gene were analyzed by genomic sequencing. There were 4 mutations in 20 metastatic melanomas. Three of 4 mutations were C:G-->T:A transitions. A search of our database of p53 mutations revealed that out of 8 p53 mutations reported by others, 4 are C:G-->T:A transitions at dipyrimidine sites, and one is a tandem CC-->TT mutation. This mutational pattern is comparable with the pattern of p53 mutations in squamous cell and basal cell carcinomas of the skin and is related to exposure to ultraviolet B (UV-B) wavelength radiation. Taken together with a predominance of UV-induced mutations in the CDKN2/ p16 gene demonstrated in melanoma cell lines, our data support a role of sunlight exposure in the etiology of malignant melanoma. The low frequency of p53 mutants in melanomas compared with other types of skin cancers suggests that although mutations in this gene are likely to be involved in the development of some malignant melanomas, they do not play as large a role as in squamous and basal cell carcinomas of the skin. PMID- 8707402 TI - Incidence of intracranial meningiomas in Nagasaki atomic-bomb survivors. AB - Among the Nagasaki atomic-bomb survivors registered at the Scientific Data Center for Atomic-Bomb Disaster, Nagasaki University School of Medicine, 45 cases of surgically treated intracranial meningioma were collected from 6 hospitals with departments of neurosurgery in or near Nagasaki City during the period from 1973 to 1992. All 45 patients were over 40 years of age at the time of diagnosis. Subsequently, the 45 cases were statistically analyzed in relationship to the estimated distance from the hypocenter by age, gender, intracranial location, histology and latent period. The analysis showed a high correlation between incidence of meningiomas and distance from the hypocenter. The incidence among Nagasaki atomic-bomb survivors over 40 years of age, especially in those proximally exposed, appears to be increasing, in inverse proportion to the exposure distance, since 1981, 36 years after the explosion of the atomic bomb. PMID- 8707403 TI - Epidemiology of childhood leukemia in Hokkaido, Japan. AB - The population-based epidemiological indices (crude incidence, survival rate, mortality, etc.) of childhood leukemia (0-14 years of age) from 1969 to 1993 in Hokkaido Prefecture, Japan, were calculated, using data obtained from the Registry of Childhood Malignancies in Hokkaido Prefecture. A total of 1,084 cases of leukemia were diagnosed in the 1969-93 period. The annual incidence of all types of leukemia from 1984 to 1993 was about 4 per 100,000 children aged 0-14 years, with the incidence of ANLL decreasing slightly and that of ALL increasing. The ratio of ALL/ANLL could similarly be seen to be increasing in all age groups. Out of a cohort of 100,000 live births, about 65 children developed leukemia by 14 years of age, and in this longitudinal observation the ratio of ALL/ANLL was increasing. The incidence of ALL and ANLL and the ratio of ALL/ANLL in Japanese children are approaching those of Caucasians. Approximately 80% of the ALL cases were of the LI type (FAB classification), and about 65% of these could be immunologically classified as "common" ALL. The 5-year survival rate of T- and B cell ALL cases was 50% or less, while that of "common" ALL cases was about 80%. PMID- 8707404 TI - Cancer incidence and mortality in women receiving estrogen and estrogen-progestin replacement therapy--long-term follow-up of a Swedish cohort. AB - We analyzed cancer incidence and mortality in a cohort of 22,597 Swedish women who were prescribed replacement hormones. After 13 years of follow-up in national registries, 2,330 incident cancer cases and 848 cancer deaths were observed. Overall, our results were reassuring since incidence rate ratios (SIRs) for 16 cancer sites and mortality ratios (SMRs) for all 10 examined sites were at, or lower than, unity. However, we found that exposure to an estrogen-progestin combined brand was associated with an increasing relative risk of breast cancer with follow-up time, the SIR reaching 1.4 (95% CI 1.1-1.8) after 10 years of follow-up. The relative risk of endometrial cancer was substantially increased, with the highest SIR of 5.0 (95% CI 1.6-5.9) in women prescribed estrogens alone, whereas those given an estrogen-progestin combination showed no elevation in risk. The risk estimates for liver and biliary tract cancers and for colon cancer were reduced by about 40%, notably in women prescribed the estradiol-progestin compound. Further detailed analyses revealed no evidence of adverse or protective effects on the risk of ovarian, uterine cervical, vulvar/vaginal, rectal, pancreatic, renal, lung, thyroid and other endocrine cancers, brain tumors, malignant melanoma or other skin cancers. Hormone replacement therapy was not associated with an increase in mortality for any cancer site, at this time of follow-up. For breast and endometrial cancers, SMRs were below baseline but tended to increase with follow-up time. We conclude that hormone replacement increases the endometrial-cancer risk after unopposed estrogens and the breast cancer risk-notably after estrogen-progestin combined therapy-and tentatively suggest that it exerts a protective effect against colon and liver cancer risks. PMID- 8707405 TI - Mechanisms of immune suppression in patients with head and neck cancer: influence on the immune infiltrate of the cancer. AB - Freshly excised human head and neck cancers (219 primary cancers; 64 metastatic lymph node cancers) were analyzed for the immune inhibitory mediators released from the cancer tissues and the immune infiltrate within the tumor. Significant levels of the immune inhibitory mediators transforming growth factor-beta (TGF beta), prostaglandin E2 (PGE2) and interleukin-10 (IL-10) were released from these cancers. Also released was granulocyte-macrophage colony-stimulating factor (GM-CSF), whose secretion was associated with an intratumoral presence of CD34+ cells. We have previously shown that CD34+ cells within human head and neck cancers are immune inhibitory granulocyte-macrophage progenitor cells. The presence of TGF-beta, PGE2 and IL-10 was associated with a reduced content of CD8+ T-cells within the cancers. The CD4+ cell content appeared to be less affected by these immune inhibitory mediators. Instead, parameters indicative of CD4+ cell function (p55 IL-2 receptor expression, release of IL-2 and IFN-gamma) were diminished in cancers that released higher levels of TGF-beta, IL-10 and GM CSF and had a higher CD34+ cell content. Furthermore, metastatic cancers released higher levels of the soluble immune inhibitory mediators and lower levels of IFN gamma and IL-2 than did primary cancers, although CD34+ cells were similarly present in both primary and metastatic cancers. Our results show that human head and neck cancers have a multiplicity of non-mutually exclusive mechanisms of immune suppression that are most prominently associated with reduced CD8+ cell influx and reduced influx and altered function of intratumoral CD4+ cells. PMID- 8707406 TI - An EWS/ERG fusion with a truncated N-terminal domain of EWS in a Ewing's tumor. AB - As a result of chromosome translocations, the EWS gene is fused to a variety of transcription factors in human solid tumors. Up to now, gene fusions of EWS with 6 different partners have been described. In all fusions presently reported the entire N-terminal domain of EWS (NTD-EWS) composed of 265 amino acids encoded by the first 7 exons of EWS was always included in the chimeric proteins, suggesting that the integrity of this domain was mandatory for the oncogenic property of the fusion proteins. We report the molecular characterization of a Ewing tumor demonstrating a reciprocal t(21;22)(q22;q12) translocation. No EWS/ERG fusion transcript could be detected with previously reported RT-PCR primers. However, Southern-blot experiments demonstrated that the EWS gene was disrupted within a 2 kb PstI genomic fragment including exon 7. PCR amplification and sequence of the translocation junction fragments indicated that the breakpoint was localized within exon 7 of EWS. The resulting fusion gene encoded a chimeric protein in which a truncated NTD-EWS was linked, in frame, to the ETS DNA-binding domain of ERG. This observation indicates that, to avoid false negative results, RT-PCR based diagnosis of tumors with EWS fusion transcripts should now include the search for such rare variants. It also suggests that the amino-terminal portion of the NTD-EWS, but not its carboxy terminal part, might be fundamental for the oncogenicity of the chimeric proteins. PMID- 8707407 TI - Cancer consequences of the Chernobyl accident in Europe outside the former USSR: a review. AB - The accident which occurred during the night of April 25-26, 1986 in reactor 4 of the Chernobyl nuclear power plant in the Ukraine released considerable amounts of radioactive substances into the environment. Outside the former USSR, the highest levels of contamination were recorded in Bulgaria, Austria, Greece and Romania, followed by other countries of Central, Southeast and Northern Europe. Studies of the health consequences of the accident have been carried out in these countries, as well as in other countries in Europe. This report presents the results of a critical review of cancer studies of the exposed population in Europe, carried out on the occasion of the 10th anniversary of the Chernobyl accident. Overall, three is no evidence to date of a major public health impact of the Chernobyl accident in the field of cancer in countries of Europe outside the former USSR. PMID- 8707408 TI - Vimentin expression is not associated with poor prognosis in breast cancer. AB - The clinical significance of vimentin intermediate filament (VIF) expression was studied in relation to other established prognostic parameters in primary breast cancer. Archival tumour samples embedded in paraffin were examined by immuno histochemistry with monoclonal antibodies (MAbs) to VIF, p53 protein and cell proliferation marker MIB-I. The vimentin staining pattern was heterogeneous, but in vimentin-positive areas > 80% of the tumour cells were positive. There was no association between vimentin expression and tumour size or the number of axillary lymph nodes involved. Vimentin expression was significantly associated with high grade tumours, absence of hormone receptors, increased p53 expression and high tumour proliferation fraction as estimated by MIB-I count. Despite these associations with several recognised features of tumour aggressiveness, vimentin expression was not associated with increase in risk of relapse or death from breast cancer. PMID- 8707409 TI - Steroid-hormone receptors in cell lines and tumor biopsies of human lung cancer. AB - Female gender is a significant independent favorable prognostic factor in lung cancer. To study the possible role of sex hormones in lung cancer, the expression of sex-steroid receptors and the glucocorticoid receptor was investigated in 29 lung-cancer cell lines stemming from small-cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC) by means of immunocytochemistry, ligand-binding assays and RNA expression via polymerase chain reaction. In at least 2 methods of investigation, NSCLC cell lines showed a low expression of estrogen receptor in 6, progesterone receptor in 13 and androgen receptor in 12 out of 17 cases examined; sex-steroid-receptor expression was virtually absent in SCLC cell lines. The glucocorticoid receptor was expressed in all 29 cell lines studied. Additionally, 52 tumor samples from primary lung cancer were investigated for their receptor expression by means of immunohistochemistry. Among patients with primary lung-cancer sex-steroid-receptor expression in tumor biopsies was detected most frequently in female patients (in 69% of 16 cases, vs. 42% of 36 tumors from men) and in patients with adenocarcinoma. Further research will focus on these subgroups. Immunohistology is a feasible method of studying steroid receptor expression in lung cancer. PMID- 8707410 TI - Natural resistance against tumors grafted into the brain in association with histocompatibility-class-I-antigen expression. AB - The role of MHC-class-I-antigen expression in intracerebral anti-tumor natural resistance was examined using MHC-positive Lym+ and MHC-negative Lym- lymphoma cell lines. Lym+ was sensitive to MHC-class-I-restricted CTL-mediated lysis, while lym- was resistant. Both lines were susceptible to NK-cell-mediated lysis. There was no difference in in vitro growth rate of in vivo intraperitoneal tumorigenicity between them. Inoculation of Lym+ cells into the brain caused upregulation of the intracellular MHC mRNA to the same level as after treatment with interferon-gamma, resulting in an increase in cell-surface MHC expression. Although inoculated Lym- cells also underwent an increase in cytosolic MHC mRNA, the cell-surface MHC expression remained negative. Immunoprecipitation revealed that the terminal glycosylation did not occur normally in Lym-. An in vivo intracerebral tumorigenicity assay, using 2 groups of untreated and NK-cell depleted syngeneic mice, showed that Lym+ was less tumorigenic than Lym-. In T cell-depleted mice, however, no difference was detected between them. In addition, when Lym+ and Lym- cells were inoculated into the brain of allogeneic or syngeneic preimmunized mice (immunized with tumor cells), Lym+ was rejected, while Lym- was accepted. When allogeneic mice had received treatment for T-cell depletion before intracerebral inoculation, no rejection was observed in Lym+. On the other hand, Lym- cells, when injected i.p. into NK-depleted mice, had greater killing activity than Lym+ cells, while in T-cell-depleted mice Lym- was less tumorigenic than Lym+. These results suggest that MHC-positive tumor cells grafted into the brain may be rejected by CTL in an MHC-dependent manner, whereas MHC-negative tumor cells can escape from T-cell-mediated immunosurveillance and grow progressively in the brain, due to absence of intracerebral natural resistance mediated by NK cells. PMID- 8707411 TI - Direct relationship between radiobiological hypoxia in tumors and monoclonal antibody detection of EF5 cellular adducts. AB - While the potential importance of hypoxia in limiting the sensitivity of tumor cells to ionizing radiation has long been appreciated, methods for accurately quantifying the number of radiation-resistant hypoxic cells within tumors have been lacking. We have used the pentafluorinated derivative [2-(2-nitro-1H imidazol-1-yl)-N-(2,2,3,3,3-pentafluoropropyl)-acet amide] of etanidazole (EF5), which binds selectively to hypoxic cells. The adducts formed between EF5 and cellular proteins in the hypoxic cells were detected using the specific monoclonal antibody (MAb), ELK3-51 conjugated to the flurochrome Cy3, and the number of hypoxic cells was quantified by flow cytometry. To verify the validity of this technique for the detection of hypoxic cells, mice bearing KHT sarcomas were treated with various agents to alter tumor oxygenation and hence vary the fraction of radiobiologically hypoxic tumor cells. The percentage of EF5 binding cells was then compared directly with the clonogenic survival of the tumor cells following radiation treatment under the various pretreatment conditions. The results showed that allowing the mice to breathe carbogen (5% CO2/95% O2) prior to irradiation reduced clonogenic cell survival approx. 6-fold and led to an absence of cells binding high levels of EF5. In contrast, pretreating the tumor bearing animals with either hydralazine, which decreased tumor blood flow, or phenylhydrazine hydrochloride, which made the mice anemic, increased tumor cell survival following irradiation 2- to 4-fold, indicative of an increase in the fraction of hypoxic tumor cells. EF5 measurements made under identical conditions illustrated a shift in the cells in the tumor to high EF5 binding. Our results demonstrate that flow cytometric measurement by fluorescent MAb binding to EF5 adducts may relate directly to radiobiological hypoxia in KHT tumors measured by conventional methods. PMID- 8707412 TI - A metastatic neuroblastoma model in SCID mice. AB - Neuroblastoma (NB), a neural crest derived tumor in children, shows a characteristic pattern of dissemination that includes adrenal glands, local lymph nodes, bone, liver, skin, and bone marrow. We have reconstructed a similar metastatic pattern in SCID mice following tail vein injection of human NB cells. HTLA230, an NB cell line isolated from a patient with advanced disease, and its NGF receptor (trkA) expressing derivative (18-10) cells, consistently disseminated to the liver, the adrenal gland, and the bone marrow, but not the lungs. Metastases in the different organs showed a characteristic hemorrhagic histopathology, and tumors in the bone marrow presented as syncytia-like cell aggregates, typically seen in patients. Cell lines reestablished from 18-10 derived liver and bone marrow metastases maintained their cellular morphology, growth behavior, N-myc overexpression, trkA expression, and functionally responded to NGF treatment, leading to growth arrest and neurite outgrowth. Hence circulating human NB cells in SCID mice show a similar organ-specific metastatic potential as seen in patients, independent of trkA expression. PMID- 8707413 TI - In vitro evaluation of a p53-expressing adenovirus as an anti-cancer drug. AB - Deficiency in p53-mediated cell death is common in human cancer, contributing to both tumorigenesis and chemoresistance. In an attempt to restore p53, we evaluated in vitro infectivity and cytotoxicity of a wild type (w.t.) p53 expressing adenovirus (Ad-p53) toward a panel of human cancer cell lines (n = 19). At a multiplicity of infection of 30, both Ad-p53 and adenovirus expressing beta-galactosidase (Ad-LacZ) infected greater than 99% of cells derived from brain, lung, breast, ovarian, colon, and prostate cancer, but failed to infect leukemia or lymphoma cells. Ad-p53, but not Ad-LacZ, infection of cancer cells was followed by nuclear accumulation of the CDK inhibitor p21WAFI/CIPI, cell cycle arrest and loss of viability. Ad-p53 induced apoptotic death in cancer cells that express mutant p53, including multi-drug resistant cells, but fewer deaths were observed in some w.t. p53 expressing cells. Ad-p53-infected SKBr3 breast cancer cells were more sensitive to cytotoxicity of the DNA damaging drugs mitomycin C or Adriamycin, but not the M-phase specific drug vincristine. Our results suggest that Ad-p53 is capable of infecting and killing cancer cells of diverse tissue origins (including multi-drug resistant cancer cells), that p21WAFI/CIPI may be a useful marker of p53 infectivity and that there may be synergy between Ad-p53 and either mitomycin C or Adriamycin induced cell death in tumors with p53 mutations. PMID- 8707414 TI - Stimulatory effects of pleural fluids from mesothelioma patients on CD44 expression, hyaluronan production and cell proliferation in primary cultures of normal mesothelial and transformed cells. AB - Cell lines established from human malignant mesotheliomas, but not from normal mesothelial cells, have been shown to possess hyaluronan receptors, and to secrete factors that stimulate hyaluronan production by fibroblasts and normal mesothelial cells. In the present study we investigated the generality of this observation, namely the presence of hyaluronan receptors and factors which induce stimulation of hyaluronan synthesis in primary mesothelioma and mesothelial cell cultures. Functionally active hyaluronan-binding sites on the surface of malignant mesothelioma cells in primary cultures, established from pleural effusions of 3 different patients, were demonstrated using 3H-hyaluronan. Primary cultures of normal mesothelial cells from non-mesothelioma effusions did not exhibit any binding ability. Pleural fluids from mesothelioma patients both stimulated hyaluronan synthesis and promoted proliferation of normal mesothelial cells to a larger extent than non-mesothelioma fluids. The hyaluronan-stimulatory activity was only slightly neutralized by antibodies against PDGF-BB or TGF-beta; antibodies against bFGF had no effect. Although the concentration of hyaluronan was much higher in pleural fluids from mesothelioma than from non-mesothelioma patients, its molecular weight was almost the same. The hyaluronan-binding capacity of early-passage mesothelioma cells derived from pleural effusions can be an additional marker, in combination with other diagnostic tools, to distinguish between mesothelioma and mesothelial cells. PMID- 8707415 TI - Alterations of the thymic selection process in transgenic mice expressing SV40 large T antigen. AB - We generated SV40 T antigen transgenic mice (lines SVT125, SVT127, and SVT248) which developed unique thymic carcinomas originating from thymic cortical epithelial cells. In these mice we observed alterations in the thymic selection process not reported before in SV40 T antigen transgenic mice. Along with tumor cell growth, thymocytes increased in number and the proportion of CD4 or CD8 single positive cells rose to 10 times the normal level. Expression of SV40 T antigen was detectable by Northern analysis in thymic stromal cells but not in thymocytes. Thymic stromal cell lines, derived from the thymic tumor, produced high levels of cytokines which caused morphological transformation and growth stimulation in hematopoietic stem cells, including fetal liver cells and bone marrow cells. These observations suggest that the unusual multiplication of thymocytes and the alterations in thymic selection are the result of the activity of thymic stromal cells transformed by SV40 T antigen. The cell lines derived from the tumor can thus be used to study cytokines involved in thymic differentiation of T cells. PMID- 8707416 TI - Polyoma tumor development in neonatally polyoma-virus-infected CD4-/- and CD8-/- single knockout and CD4-/-8-/- double knockout mice. AB - CD4-/- or CD8-/- single knockout as well as CD4-/-8-/- double knockout mice were infected with polyoma virus as newborns or 1 week after birth. The animals were followed for tumor development and virus persistence. Double knockout mice developed tumors at a higher incidence (29%) than either the CD8-/- or CD4-/- single knockout mice (11% and 2%, respectively). Persistence of polyoma virus was examined by PCR in one third of all animals included in the study. Seven of the 17 CD4-/-8-/- double knockout mice gave positive evidence of virus persistence up to 6 months p.i. where virus DNA was present in most organs. Corresponding tests in single knockout mice gave positive results of persistent viral DNA in 2 of the 19 CD8-/-and 2 of the 7 CD4-/-mice. In the single knockout mice polyoma DNA could only be detected in a more limited variety of organs compared to the double knockouts. PMID- 8707417 TI - Different ligand responsiveness of human retinoic-acid-receptor beta-gene transcription in tumorigenic and non-tumorigenic cervical-carcinoma-derived cell lines is mediated through a large retinoic-acid-response domain. AB - The retinoic-acid-receptor beta gene (RAR-beta) encodes a suspected tumor suppressor for several types of human carcinomas. RAR-beta transcription is induced by retinoic acid (RA) through retinoid receptors which bind as heterodimers of a RA-activated RA receptor (RAR) and a retinoid X receptor to the RA-responsive element in the RAR-beta promoter region (beta RARE). RA inducibility of RAR-beta gene expression is often lost or reduced in human carcinoma cells. As previously shown, the RAR-beta gene is highly RA-inducible in nontumorigenic HeLa x fibroblast hybrid cells, but neither in HeLa cervical carcinoma cells nor in a tumorigenic hybrid segregant line. We report here that severe reduction of RA-induced RAR-beta mRNA levels is a general feature of tumorigenic HeLa x fibroblast segregants. To study the molecular basis of differential RA inducibility, we have performed transient transfection assays in HeLa and nontumorigenic 444 hybrid cells using reporter constructs with different 5' and internal deletions of the RAR-beta transcription-control region. Remarkably, maximal RA inducibility in 444 cells required the integrity of the complete RAR-beta upstream region. In HeLa cells, all reporter constructs showed only low RA inducibility levels. The differential RA inducibility in 444 and HeLa cells could be conferred by the RAR-beta upstream region, but not by subfragments of it, on a heterologous RA-responsive promoter. The data indicate that maximal RA inducibility of RAR-beta gene transcription in nontumorigenic 444 cells depends on the cooperation of the beta RARE with additional upstream elements. All elements together constitute a large RA response domain as the higher-order transcription control unit. The communication between the upstream elements and the beta RARE seems to be disturbed in HeLa cells. Similar defects may be responsible for the loss of RA responsiveness of RAR-beta gene expression in other human tumors. PMID- 8707418 TI - Monoclonal antibodies against recombinant-MAGE-1 protein identify a cross reacting 72-kDa antigen which is co-expressed with MAGE-1 protein in melanoma cells. AB - The MAGE-1 gene codes for tumor-associated peptides recognized by cytolytic T lymphocytes in association with MHC-class-1 molecules such as HLA-A1 and HLA Cw16. In the course of a study aiming at the immunohistochemical detection of the MAGE-1 gene product in tumor samples, 2 mouse monoclonal antibodies (MAbs) directed against a full-length recombinant MAGE-1 fusion protein were found to react strongly not only with the 46-kDa MAGE-1 protein, but also with a 72-kDa product in immunoblots of lysates obtained from several MAGE-1-mRNA-positive melanoma cell lines. Pre-incubation of the antibodies with the recombinant MAGE-1 fusion protein abolished their reactivity both with MAGE-1 protein and with the 72-kDa product, thus confirming the occurrence of antigenic determinant(s) shared by the 2 proteins. The 72-kDa protein is not an alternative product of MAGE-1, since it was still detected in lysates of a MAGE-1 loss variant derived from a MAGE-1-positive melanoma cell line. Moreover, the 72-kDa protein does not appear to be a product of the other members of the MAGE gene family known to be expressed in tumors (such as MAGE-2, -3, -4 and -12). Interestingly, expression of the 72-kDa protein was found to be correlated with that of MAGE-1 protein. Thus, in 30 tumor cell lines analyzed by immunoblotting and RT-PCR, the 72-kDa protein was never detected in MAGE-1-mRNA-negative cell lines, while it was co expressed with MAGE-1 protein in 12 out of 15 cell lines expressing MAGE-1. Furthermore, the 72-kDa protein was detected in lysates of human testis, the only normal tissue known to express MAGE-1. Finally, treatment of MAGE-1-mRNA-negative cell lines with 5-Aza-2'-deoxycytidine, a hypomethylating agent known to induce MAGE-1 expression, resulted in the expression of the 72-kDa protein. Taken collectively, these findings suggest that expression of the gene encoding the 72 kDa protein identified in this study through antigenic determinant(s) shared with MAGE-1 protein is regulated in a way similar to that of MAGE-1. PMID- 8707419 TI - Overexpression of urokinase receptor in breast cancer cells results in increased tumor invasion, growth and metastasis. AB - We have examined the role of urokinase receptor (uPAR) in tumor invasion and metastasis by developing a homologous model of uPAR overexpression in a rat breast cancer cell line (Mat B III) using gene transfer technique. Control (pRc CMV) and experimental plasmid (pRc-uPAR-S) were transfected into Mat B III cells by using Lipofectin reagent. Levels of uPAR production were accessed by Northern blotting, immunofluorescence, receptor binding and ELISA. At least 3 experimental clones (pRc-uPAR-S), expressing 3- to 5-fold higher levels of uPAR than control (pRc-CMV), were selected for further analysis. Experimental cells overexpressing uPAR showed a 4-to 5-fold higher invasive capacity compared with control cells in a Boyden chamber invasion assay. Both control and experimental cells (1 x 10(6) cells) were injected into the mammary fat pad of syngeneic female Fischer rats. Animals were sacrificed at timed intervals and evaluated for the development of tumor growth and metastasis. Animals receiving cells overexpressing uPAR had significantly larger tumor volume and weight throughout our study. Furthermore, due to increased uPAR expression, experimental animals developed large metastatic lesions in liver, spleen and lymph nodes. Our results therefore demonstrate the role of uPAR in tumor progression, due to its ability to localize uPA within the tumor cell milieu. PMID- 8707420 TI - Susceptibility of human melanoma cells to oxidative stress including UVA radiation. AB - Ultraviolet radiation, and in particular UVA (320-400 nm), induces significant oxidative stress to human skin. Ferritin and glutathione have been shown to be among the more important molecules within human skin cells providing protection against this damage, the presence of lower levels of these anti-oxidants giving rise to increased cellular sensitivity to stress. We compared endogenous levels of ferritin and glutathione in human melanoma cells with normal human skin fibroblasts and keratinocytes, also the response of melanoma cells to oxidative stress with fibroblasts and keratinocytes. Ferritin levels were heterogenous in the untreated melanoma cell lines tested and remained the same following oxidative stress (UVA radiation) or hemin treatment. Epidermal keratinocytes were unaffected, as were the melanoma cell lines, but skin fibroblasts showed dose dependent ferritin depletion. Similar results were seen for glutathione alterations resulting from UVA radiation: melanoma cell lines and epidermal skin keratinocytes remained unchanged following UVA radiation, while skin fibroblasts showed dose-dependent depletion. Our results show that human melanoma cells have low ferritin and glutathione levels, yet are resistant to oxidative stress. PMID- 8707421 TI - Potentiation of the reversal activity of SDZ PSC833 on multi-drug resistance by an anti-P-glycoprotein monoclonal antibody MRK-16. AB - SDZ PSC833 (PSC833), an analogue of cyclosporines, is one of the most potent modulators of multi-drug resistance (MDR). We previously reported that MRK-16, an anti-P-glycoprotein MAb, enhanced MDR reversal activity of cyclosporin A (CsA) through inhibition of P-glycoprotein-mediated CsA transport. We have examined here whether MRK-16 can enhance MDR reversal activity of PSC833. We found that MRK-16 potentiated the MDR reversal activity of PSC833, and of CsA, in MDR sublines of human myelocytic leukemia K562 and human ovarian cancer A2780 cells. Like MRK-16 combined with CsA, MRK-16 enhanced the effect of a sub-optimum dose of PSC833 on vincristine accumulation in MDR cells. However, MRK-16 could not increase cellular accumulation of PSC833 in MDR tumor cells, yet it could increase cellular accumulation of CsA. P-glycoprotein could not transport PSC833 but could transport CsA. Our results indicate that MRK-16 potentiates the MDR reversal activity of both PSC833 and CsA, yet also suggest that the molecular mechanism of the potentiation differs between the two substances. PMID- 8707422 TI - Oncogene expression is modulated by recombinant human interferon-beta in human breast-cancer cells. AB - The effect of recombinant human interferon-beta on growth and oncoprotein expression was investigated in several human breast-cancer cell lines with different characteristics. All cell lines tested were sensitive to the antiproliferative action of the drug, regardless of their estrogen sensitivity. The maximal inhibition of cell proliferation was seen after 6 days of treatment. In estrogen-sensitive CG-5 and ZR-75-1 cells, but not in MDA-MB-453 estrogen insensitive cells, a reduction in c-myc and c-erbB2 oncoproteins occurred after 48-72 hr and became more pronounced after 120-168 hr of treatment, suggesting that this down-regulation is not direct but is mediated by undefined molecular mechanisms. The time-course of the IFN-mediated decrease in oncoproteins seems to indicate that this event is not strictly related to the IFN-regulation of cell proliferation. The expression of c-erbB2 and c-myc was also analyzed, after recombinant human interferon-beta treatment, at the mRNA level in CG-5 cells. Surprisingly, no statistically significant variation of c-erbB2 or of c-myc mRNA was found either before or after 120-168 hr. Thus, we surmise that the observed reduction of oncoproteins may be due to post-transcriptional mechanisms. PMID- 8707423 TI - Clonality and stability of the p53 gene in human astrocytic tumor cells: quantitative analysis of p53 gene mutations by yeast functional assay. AB - Mutation of the p53 gene is found in about one third of astrocytic brain tumors, and expansion of tumor cell clones containing mutant p53 has been implicated in astrocytic tumor progression. However, admixture of normal cells in astrocytic tumor specimens limits the power of traditional studies of tumor cell clonality. To address this problem we have employed a yeast p53 functional assay that scores the content of mutant p53 alleles in tumors and cell lines quantitatively. We have analyzed 17 cases where matching tumor material and derived cell lines were available. The yeast assay gave > 20% red (i.e., mutant p53-containing) yeast colonies in 7 out of 17 cases. One case had no mutations in the primary tumor but gave 76% red colonies in a recurrence, clearly demonstrating tumor overgrowth by a mutant clone. During early passages of cultured tumor cells, mutant p53 content increased rapidly with passage due to outgrowth of mutant clones from a heterogeneous starting population. In addition, de novo p53 mutations appeared during culture in 2 cases. This indicates that there is stronger selective pressure for mutation during the establishment of cell lines in vitro than during tumor growth in vivo. Our results demonstrate the utility of the p53 functional assay for studies of clonality and support the hypothesis of clonal progression of brain tumors in vivo. PMID- 8707424 TI - Expression of the endogenous retrovirus ERV3 (HERV-R) during induced monocytic differentiation in the U-937 cell line. AB - ERV3 (HERV-R) is a complete human endogenous retrovirus located on the long arm of chromosome 7. LTR-env-gene-spliced mRNA of 9 and 3.5 Kb is widely expressed in human tissues and cells, but gag-pol mRNA has not been found. Further, the env gp70 gene contains an open reading frame throughout its length and its expression has recently been detected as a full-length protein. The highest expression of ERV3 detected so far is in placenta and the lowest in cytotrophoblasts and choriocarcinoma cell lines. In this report we have studied ERV3 mRNA and protein expression in the human monoblastic cell line U-937 during differentiation into monocytes/macrophages. Differentiation of U-937 cells was induced by 1,25a dihydroxyvitamin D3 (vitD3), retinoic acid (RA), gamma interferon (IFN-gamma) and phorbol-myristate-acetate (PMA-TPA). The expression of ERV3 env mRNA was found to be differentiation-associated, with high expression detected in the late stages of monocytic development. Using TPA, the expression of ERV3 env was detected as 9 and 3.5-kb transcripts by Northern blotting, as mRNA by in situ hybridization and as a cytoplasmic 65-kDa protein by immunofluorescence and Western blots. Low levels of basal expression were found, with up-regulation of both message and protein at 24 to 48 hr after addition of TPA. Induction with vitD3, IFN-gamma and RA produced higher levels of mRNA at earlier time points. It is concluded that the U-937 cell line represents an excellent model system for further studies to study the relationship between ERV3 expression and cellular differentiation. PMID- 8707425 TI - MAGE BAGE and GAGE genes expression in fresh epithelial ovarian carcinomas. PMID- 8707426 TI - The ratio of free to total prostate-specific antigen in serum is correlated to the prostate volume. PMID- 8707427 TI - Sudden cardiac death in hypertrophic cardiomyopathy: risk evaluation. AB - Mortality due to hypertrophic cardiomyopathy is only around one percent per annum in unselected patients [1]. Therefore, the incidence of sudden cardiac death is still lower. However, identification of this small subgroup is important because prophylactic intervention may reduce mortality [2-5]. Unfortunately, we still do not have an accurate method of identifying this high risk subgroup. PMID- 8707428 TI - Rotational atherectomy of calcified ostial saphenous vein graft lesion with long term follow-up: a case report. AB - Rotational atherectomy, though widely used for calcified and difficult lesions, has not been used in vein graft lesions. We performed rotational atherectomy on a 75-year-old male with a calcified ostial lesion in an 11-year-old vein graft. No complications were encountered and good result was obtained. Angiogram at 10 months revealed no restenosis. PMID- 8707429 TI - Concurrent aortic and mitral balloon valvuloplasty by the retrograde non transseptal technique. AB - Balloon valvuloplasty for combined aortic and mitral stenosis is now being increasingly practised. Transseptal catheterisation forms an integral part of this procedure. We describe a case where the retrograde non-transseptal approach was used for dilating both valves in a single intervention. PMID- 8707430 TI - Beneficial cumulative role of both nitroglycerin and dobutamine on right ventricular systolic function in congestive heart failure patients awaiting heart transplantation. AB - End-stage idiopathic dilated cardiomyopathy or ischemic heart disease usually present with very low cardiac output and severe ventricular dysfunction which may require pharmacological support before heart transplantation. Right ventricular ejection fraction might be an important factor of functional capacity and survival in congestive heart failure. In order to test the immediate response of right ventricular hemodynamic parameters to nitroglycerin and dobutamine usually used to treat severe left ventricular dysfunction, we studied 17 congestive heart failure patients (15 men, two women; mean age 55 +/- 13 years) with end-stage idiopathic dilated cardiomyopathy (n = 10) or end-stage ischemic heart disease (n = 7), left ventricular ejection fraction < 35% (mean 22 +/- 8%), and sinus rhythm. A well validated thermodilution technique using a dedicated catheter with a fast catheter-computer response, permitting instantaneous measurements of right ventricular ejection fraction, was used. Right ventricular hemodynamic parameters were recorded at baseline, after an intravenous bolus injection of 3 mg nitroglycerin and after an intravenous infusion of dobutamine administered after nitroglycerin until normalization of cardiac index or a maximal dose of 15 micrograms/kg/min. Pulmonary artery mean pressure significantly decreased after nitroglycerin (43 +/- 9 mmHg at baseline vs. 31 +/- 10 mmHg after nitroglycerin, P < 0.0001) and did not subsequently change after dobutamine (32 +/- 10 mmHg after dobutamine, ns). Cardiac index was not affected by nitroglycerin (1.7 +/- 0. l/min/m2 at baseline vs. 2.0 +/- 0.3 l/min/m2 after nitroglycerin, ns), but dramatically increased after dobutamine (3.0 +/- 1.0 l/min/m2 after dobutamine, P < 0.0001). Concomitantly to the changes of these two parameters, right ventricular ejection fraction progressively increased (14 +/- 8% at baseline vs. 20 +/- 10% after nitroglycerin (P < 0.0006) vs. 28 +/- 13% after dobutamine (P < 0.0001)). Progressive increase of right ventricular ejection fraction after administration of nitroglycerin followed by administration of dobutamine suggests the beneficial cumulative role of both medications on right ventricular systolic function in severe congestive heart failure. PMID- 8707431 TI - Obesity and left ventricular diastolic dysfunction. AB - To assess the influence of obesity on left ventricular function, 20 obese women (mean body mass index (BMI) 33.8 +/- 3.1 kg/m2 and mean age 31.1 +/- 2.4 years) without evidence of heart disease were evaluated by echocardiography. Obese subjects had greater left ventricular mass index (103 +/- 22 g/m2, 76 +/- 18 g/m2; P < 0.0001) and augmented fractional shortening (39 +/- 2.6%, 36 +/- 0.1%; P < 0.0001) than normals. Isovolumic relaxation time was prolonged in the obese group (92 +/- 11 ms) as compared with the control group (76 +/- 11 ms; P < 0.0001). The ratio of peak early and atrial filling velocities was significantly lower (1.2 +/- 0.4, 1.9 +/- 0.6; P < 0.0001) and atrial contribution was higher (39 +/- 9, 25 +/- 5; P < 0.0001) in obese subjects than in normals. Shortened deceleration time was measured in obese subjects (142 +/- 30, 179 +/- 20 ms; P < 0.0001). In conclusion, obesity causes relaxation and early filling abnormalities and diastolic filling is compensated by augmented atrial contribution. Diastolic dysfunction is an early indicator of cardiac involvement in obesity. PMID- 8707432 TI - QT prolongation and polymorphous ventricular tachycardia associated with trasodone-amiodarone combination. AB - Trasodone, a second generation antidepressant, has been shown to equal standard tricyclics in clinical efficacy while causing significantly fewer cardiac side effects. Although clinical trials have demonstrated a relatively safe arrhythmogenic profile, trasodone has been implicated in the pathogenesis of ventricular arrhythmias. This report describes a patient in whom the administration of trasodone in addition to previously well-tolerated long-term amiodarone therapy was associated with a marked prolongation of the QT interval and polymorphous ventricular tachycardia. Caution in the concurrent use of trasodone and other medications known to cause QT prolongation and polymorphous ventricular tachycardia is recommended. PMID- 8707433 TI - Chronobiological aspects of pulmonary thromboembolism. AB - A growing body of evidence has recently accumulated suggesting that several unfavorable cardiovascular events, e.g., myocardial angina and infarction, sudden cardiac death, and stroke, present a specific temporal pattern in their occurrence. This has also been shown to be true for fatal pulmonary thromboembolism, with a high frequence peak in winter and in the morning hours. The authors report the circannual and circadian aspects of pulmonary thromboembolism, analysing both the temporal patterns of the underlying favouring or precipitating factors, and the possible therapeutic implications. PMID- 8707434 TI - Prevalence of hypertension and its trends in Chinese populations. AB - The third nation-wide survey of blood pressure level and hypertension was carried out in China in 1991. In total, 950,356 residents aged > or = 15 years were examined. Sampling population were composed of half urban and half rural from 27 provinces or autonomous regions and three municipalities. The results showed that: (1) the age-adjusted prevalence rate of hypertension (systolic blood pressure > or = 140 mmHg or diastolic blood pressure > or = 90 mmHg) was 11.26% (male 12.15%, female 10.32%) and the definite hypertension rate (systolic blood pressure > or = 160 mmHg or diastolic blood pressure > or = 95 mmHg, or on medication) was 5.29% (male 5.38%, female 5.21%); rate of borderline hypertension was evaluated from 2.82% to 6.15% during the 10-year period from 1979/1980 to 1991; (2) the prevalence rate was progressively increased with age, especially after age 35; prevalence rates were generally higher in men than women before about age 44, however by age 60, women had a higher prevalence of hypertension; (3) a general trend of decrease in prevalence from north-eastern to south-western China was confirmed; (4) there were significantly lower rates in rural than in urban areas. Results of preliminary analysis for prevalence of hypertension among different occupations, nationalities, and educational levels were presented. PMID- 8707435 TI - Magnetic resonance imaging of vascular changes in Takayasu arteritis. AB - We studied the thoracic aorta in 42 female patients with Takayasu arteritis by magnetic resonance imaging (MRI) with a 1.5-T superconducting instrument and compared the findings with those in 10 healthy female controls. Images in the transaxial and oblique sagittal planes were acquired by cardiac gating using a spin-echo pulse sequence. Images in the oblique plane were also obtained with cine MRI. Vascular changes were found in 41 (98%) of the patients. Dilatation of the ascending aorta was observed in 20 patients (48%), while stenotic changes were more characteristic in the descending aorta (16 patients, 38%). Moderate signal-intensity masses on the aortic wall were encountered in 21 patients (50%), suggesting mural thrombi. Turbulent blood flow was observed around all masses by cine MRI. HLA analysis revealed that patients carrying HLA-B52 typing showed these masses more frequently than those without this typing (P < 0.05). Cine MRI demonstrated aortic regurgitation in 20 patients (48%), all of whom showed dilated ascending aorta; furthermore, 14 (70%) of them were confirmed as carrying HLA-B52. Thus, MRI was a useful non-invasive tool not only for the evaluation of the morphological changes in the aorta and its branches, and of the hemodynamic disturbance in Takayasu arteritis, but also for follow-up of the morbid condition. PMID- 8707436 TI - Balloon mitral commissurotomy in patients with previous annuloplasty. Our experience. AB - Balloon mitral commissurotomy is an alternative to surgical commissurotomy in the treatment of mitral stenosis and different studies have shown its usefulness for restenosis following surgical commissurotomy. We describe our experience in balloon mitral commissurotomy in five patients with previous commissurotomy and annuloplasty. Among 360 balloon commissurotomies performed in our hospital up to December 1993, 29 procedures were performed on patients with restenosis after surgical commissurotomy, five of whom also had an annuloplasty (flexible ring in four and rigid in one). The balloon commissurotomy was performed without complications using the Inoue single balloon technique. Mitral valve area increased from 0.9 +/- 0.1 cm2 to 1.0 +/- 0.1 cm2 by pressure half-time, and from 1.0 +/- 0.2 cm2 to 1.3 +/- 0.1 cm2 by Gorlin formula. Two patients in functional class III underwent mitral valve replacement, 32 and 11 months later; the other three patients were in class II 38, 10 and 7 months later. The presence of a mitral ring should not constitute a contraindication to balloon commissurotomy. However, the haemodynamic results are not favourable in our patients, probably due to the practical absence of commissural fusion and the ring's restrictive effect on valvular stretching. PMID- 8707438 TI - Atrial septal aneurysm. Morphological characteristics in a large population: pathological associations. A French multicenter study on 259 patients investigated by transoesophageal echocardiography. AB - A strong association between interatrial septal aneurysm (IASA) and stroke has recently led many authors to consider IASA as a potential cardiac source of embolism. We studied the morphological characteristics and main associations of IASA in a large cooperative study based on transoesophageal echocardiographic examinations; 259 IASA were studied in 134 men and 125 women with a mean age of 59 +/- 15 years. Fifty-five percent of IASA were found to overlap the commonly described fossa ovalis region. IASA protruded into the right atrium in 90% of the cases. They appeared thin in 81% of the patients and highly mobile in 79%. Fifty eight percent of patients had a history of systemic embolic events, while an atrial septal shunt was detected in 61% of the patients. In patients with an embolic event, only the mobility of IASA was significantly higher than in those with no embolic event. In nine cases a pulmonary embolism was associated with arterial embolism. Furthermore, we reported three cases of paradoxical embolism. However, the true demonstration of a thrombus within the IASA was quite rare. CONCLUSION: IASA is probably an important risk factor for stroke. In patients with IASA and a history of embolic events, IASA may enhance migration of a thrombus constituted in situ or transiting through it. Marked mobility of IASA may also increase the risk of peripheral embolus. PMID- 8707437 TI - Natural evolution of antimyosin scan and cardiac function in patients with acute myocarditis. AB - Indium-111 monoclonal antimyosin antibody imaging is a convenient method to diagnose acute myocarditis. The present study examined the natural evolution of a positive antimyosin scan in 10 patients with acute myocarditis, in relation to left ventricular ejection fraction. All patients underwent myosin-specific monoclonal antibody imaging and radionuclide ventriculography; in all patients endomyocardial biopsy was performed. Repeat assessment of left ventricular function and antimyosin uptake was performed 1, 2 and 6 months later. A heart-to lung ratio was used to quantify antimyosin uptake. Heart-to-lung ratio in patients with myocarditis was 2.12 +/- 0.31, significantly higher than that observed in eight controls (1.35 +/- 0.08, P < 0.001). During follow-up 2.2 repeat scans per patient were performed. Heart-to-lung ratio normalized (< 1.6) within 1 month in four patients, within 2 months in one patient and within 6 months in two patients; three patients continued to have a positive antimyosin uptake at 6 months, suggesting ongoing myocardial necrosis. At the end of follow up, ejection fraction improved by > 5% in four patients; normalization or persistence of tracer uptake could not predict an improvement of ejection fraction. It is concluded that antimyosin scintigraphy is useful for diagnosing myocarditis in its subacute or chronic stage during which unknown or immune mechanisms may be in operation. An improvement in left ventricular function cannot be predicted on the basis of persistence or not of tracer uptake. PMID- 8707439 TI - Balloon mitral valvotomy: an autopsy study. AB - The autopsy findings in 20 patients who died following percutaneous balloon mitral valvotomy are reported. The procedure was attempted in 508 patients. In 17 of the 20 patients, balloon mitral valvotomy was attempted as a salvage procedure. Ten patients died in the immediate post valvotomy period (within 24 h), seven died within a week and three between 1 to 3 months. In 13 patients, the balloon valvotomy successfully opened out one or both commissures. Interatrial septostomy defects created by the transeptal catheter could be identified in 19 of 20 heart specimens. The size of the defects ranged from small (5 mm) openings to large (15 mm) defects especially with double balloon. Marked mitral valvar thickening with nodular calcification was observed in 16 cases. Significant complications leading to mortality included cardiac tamponade (five cases) due to left ventricle apical perforation (3/5 cases). Mitral valve damage in the form of leaflet tears, chordal rupture and long splits in five cases resulted in significant mitral regurgitation. The leaflet tears resulted in detachment of part of the leaflet from the annulus. A late complication noted was infective endocarditis. Associated pulmonary tuberculosis, chronic obstructive pulmonary disease, respiratory infections and multivalvar diseases also contributed to mortality. PMID- 8707441 TI - A reversible dilated cardiomyopathy due to thyrotoxicosis. AB - Thyrotoxicosis gives rise to a high output cardiac failure. Rarely, it can cause a dilated cardiomyopathy with severe impairment of myocardial function which improves significantly following treatment. PMID- 8707440 TI - Immunoglobulin E, mast cell-specific tryptase and the complement system in sudden death from coronary artery thrombosis. AB - We tested the hypothesis of anaphylaxis as a pathogenic factor in acute myocardial infarction. Mast cells were counted in the myocardium, coronary arteries and airways. Total serum IgE, mast cell tryptase, complements C3, C4, and factor B, were measured in 29 cases of sudden death from coronary artery thrombosis and in 27 controls. We found increased numbers of mast cells in the coronary arteries following coronary death: 46 +/- 21 (SD) compared with 22 +/- 10 (SD) in the control group (P < 0.002). In the myocardium and airways there were no differences between the groups. The concentrations of tryptase and IgE in serum did not differ between the groups although 20% of the coronary deaths had elevated values (> 200 kU/l) compared with 8% in the control group. Of the complement factors, C3 was higher in the coronary deaths (P < 0.05) than in the controls. The results give no evidence of anaphylactic reaction in the pathogenesis of acute myocardial infarction. PMID- 8707442 TI - Abnormalities of autonomic nervous system in hyperadrenergic orthostatic hypotension: description of a case. AB - Hyperadrenergic orthostatic hypotension was diagnosed in a 27-year-old man because of recurrent episodes of hypotension associated with high plasma noradrenaline levels. In this patient, laboratory tests were performed to evaluate autonomic nervous system function. Decreased response to Valsalva maneuver and carotid sinus massage indicated decreased baroreflex and vagal responsiveness, respectively. Cardiovascular response to the handgrip was reduced in comparison to controls. Passive leg raise showed normal reduction of plasma norepinephrine, indicating normal responsiveness of cardiopulmonary receptors. 'Non-dipper' profile in the ambulatory blood pressure monitoring provided further evidence for an impaired autonomic control of cardiovascular function in this patient. This report suggests the presence of autonomic dysfunction in hyperadrenergic orthostatic hypotension. PMID- 8707443 TI - Left ventricular aneurysm as very slow complication of myocardial contusion in an athlete. PMID- 8707444 TI - The specific type IV phosphodiesterase inhibitor rolipram combined with adenosine reduces tumor necrosis factor-alpha-primed neutrophil oxidative activity. AB - Monocytes and macrophages produce tumor necrosis factor-alpha (TNF alpha) in response to microbial products including endotoxin. TNF alpha is a potent primer of neutrophil (PMN) oxidative activity. Certain xanthine phosphodiesterase (PDE) inhibitors such as pentoxifylline have been shown to inhibit stimulated oxidative activity in PMN. In the present study, the non-xanthine PDE type IV inhibitor rolipram (4-[3'-cyclopentyloxy-4'-methoxyphenyl]-2-pyrrolidone) alone and in combination with adenosine is examined as a potential modulator of TNF alpha primed PMN oxidative activity. Attainable in vivo concentrations of rolipram and physiological concentrations of adenosine alone and together synergistically decreased rhTNF alpha-primed suspended PMN oxidative activity stimulated by the chemoattractant f-met-leu-phe. The rolipram effect was reversible by washing, and rolipram had a comparable effect if added before or after priming, indicating that its effect was on the primed response rather than on priming per se. In addition, rolipram especially when combined with adenosine, decreased rhTNF alpha stimulated PMN adherence to a fibrinogen-coated surface, and the oxidative burst of rhTNF alpha-stimulated adherent PMN. The specific adenosine A2a receptor agonists CGS 21680 and WRC-0474 had comparable activity to adenosine in these experiments. Adenosine (or CGS 21680) combined with rolipram synergistically increased f-met-leu-phe-stimulated PMN cAMP content. The effects of both adenosine and rolipram with adenosine could be only partly counteracted by treatment of the PMN with the protein kinase A inhibitor KT 5720, indicating that protein phosphorylation is only partially involved. Rolipram activity was about 1000 x (by molar concentration) greater than pentoxifylline in comparable assays. Thus, rolipram, especially when combined with adenosine, has potent modulating effects on PMN activation and may be useful in decreasing inflammatory tissue damage in patients with sepsis. PMID- 8707445 TI - Cytokine-modulating activity of tepoxalin, a new potential antirheumatic. AB - Tepoxalin is a new dual cyclooxygenase/5-lipoxygenase anti-inflammatory compound currently under clinical investigation. It has been shown to possess anti inflammatory activity in a variety of animal models and more recently to inhibit IL-2 induced signal transduction. The current study was conducted to evaluate the cytokine modulating activity of tepoxalin and the role of iron in these effects. In human peripheral blood mononuclear cells (PBMC) stimulated with OKT3/PMA, tepoxalin inhibited lymphocyte proliferation with an IC50 of 6 microM. Additionally, it inhibited the production of LTB4 (IC50 = 0.5 microM) and the cytokines IL-2, IL-6 and TNF alpha (IC50 = 10-12 microM). Cytotoxicity was not demonstrated at these concentrations. Add-back experiments with either cytokines (IL-2 or IL-6), LTB4 or conditioned media failed to restore the proliferative response in the presence of tepoxalin. However, the concurrent addition of iron (in the form of ferrous or ferric chloride and other iron salts) reversed the inhibition of proliferation caused by tepoxalin. Tepoxalin also inhibits the activation of NF kappa B, a transcription factor which acts on several cytokine genes. Tepoxalin's effect on NF kappa B is also reversed by the addition of iron salts. These data suggest that the action of tepoxalin to inhibit proliferation in PBMC may be at least in part due to its ability to reduce the amount of available iron resulting in decreased activation of NF kappa B and subsequent inhibition of cytokine production. PMID- 8707446 TI - Effect of serotonin on the chemiluminescence response of rat peripheral blood leucocytes. AB - Serotonin (5-HT) has been shown to exert various immunomodulatory effects. In this study, the effects of 5-HT, 5-hydroxy-DL-tryptophan (5-HTP) and DL-p chlorophenylalanine (PCPA) on the chemiluminescence (CL) responses of rat peripheral blood leucocytes (PBL) activated by phorbol myristate acetate (PMA), opsonized zymosan or latex beads were assessed. The CL responses were measured following in vitro treatment with 0.01-100 microM 5-HT, and either 1 h after the last i.p. administration of 5-HT (0.05, 0.5, 1, 2.5, 5 or 10 mg/kg for 4 days), 5 HTP (25 or 100 mg/kg for 4 days) or PCPA (200 mg/kg for 4 days, n = 5), or 48 h after a single 200 mg/kg PCPA injection. A concentration-dependent decrease in CL responses was noted following in vitro 5-HT treatment. In vivo treatment of rats with 5-HT produced a reverse bell-curve inhibiting effects on the CL response with a maximal inhibition in rats receiving 1 mg/kg/day 5-HT and a weaker response of PMA-activated PBL. In vivo treatment with high-dose 5-HTP increased CL response of opsonized zymosan-activated PBL, while low-dose 5-HTP decreased CL response of opsonized zymosan and latex beads-activated PBL. No effect was observed in PMA-activated PBL from rats treated with 5-HTP. By contrast, in vivo treatment with PCPA increased CL responses induced by PMA or latex beads, whereas CL responses using opsonized zymosan were not significantly affected. These results suggest that 5-HT modulates the CL response of rat leucocytes to particulate stimuli. PMID- 8707447 TI - Immunotherapy with natural interleukins and/or thymosin alpha 1 potently augments T-lymphocyte responses of hydrocortisone-treated aged mice. AB - Cytokines and thymic hormones are thought to play critical roles in the regulation of T-lymphocyte development and function. In an effort to determine the effectiveness of such agents in an immunotherapeutic strategy, we employed aged mice in a hydrocortisone treatment model to generate an immunodeficient state and to study its reconstitution. Mice were given five daily injections of a natural cytokine mixture (NCM), recombinant interleukins (rIL-1, rIL-2) or their combination, thymosin alpha 1 or fraction 5 (T alpha 1, TF5), or the combinations of NCM plus T alpha 1 and of NCM plus TF5. Spleen and thymus weights were obtained and the cellular responses to stimulation in vitro with NCM, IL-1, IL-2 and mitogens (PHA and Con A) were assayed. Both NCM and T alpha 1 in vivo treatment augmented thymocyte and splenocyte in vitro responses to both interleukins and mitogens. Neither treatments with equivalent doses of rIL-1, rIL 2 nor their combination, nor TF5 achieved similar results. Of all the treatments, only NCM plus T alpha 1 augmented spleen weight; none augmented thymus weight. Surface marker analyses of T-lymphocytes and subsets indicate that treatment of mice with NCM plus T alpha 1 increased spleen T-cell numbers of both CD4 and CD8 positive cells significantly. These data indicate that NCM and T alpha 1 alone and in combination may be therapeutically useful to restore T-lymphocyte number or function in secondary immunodeficiency. PMID- 8707448 TI - Preclinical evaluation of the ribosome-inactivating proteins PAP-1, PAP-S and RTA in mice. AB - In a preclinical mouse model the plant ribosome-inactivating proteins (RIPs) pokeweed antiviral proteins PAP-1, and PAP-S and ricin A-chain (RTA) induced a pathological elevation of serum concentrations of glutamate pyruvate transaminase (GPT) and blood urea nitrogen (BUN) and had a significant immunosuppressive effect on B- and T-lymphocytes. The present analysis and comparison of the biodistribution and systemic/organ toxicity associated with RIP injection suggest a possible in vivo mechanism of action of PAP-1 and PAP-S and identify several limitations in the clinical use of these two toxins and RTA. When administered intravenously, PAP-1 and PAP-S consistently accumulated in kidneys and induced histologically documented damage to kidney and liver, with a LD50 of 3.3 mg/kg and 1.6 mg/kg for PAP-1 and PAP-S, respectively. In mice injected with PAP-S after chlorpromazine (CPZ) administration, GPT levels returned to normal between 24 and 72 h after toxin injection, while the BUN levels remained elevated. Mortality of the animals was delayed but all mice eventually succumbed. All the three toxins inhibited the expansion of anti-sheep red blood cells (SRBC) antibody-forming cells and the production of anti-SRBC antibody levels, although PAP-S showed the most potent activity. Despite the immunosuppressive activity, all toxins were highly immunogenic. PMID- 8707449 TI - Tumor necrosis factor production by human T-cells stimulated with bacterial superantigens. AB - Tumor necrosis factor (TNF) production from T-cells stimulated with superantigenic exotoxins, staphylococcal enterotoxin B and streptococcal pyrogenic exotoxin A was investigated in the presence of cells bearing distinct isotypes of HLA class II molecules. The main T-cell subset for TNF production was investigated in parallel. Similarly high levels of TNF production were induced upon stimulation with the toxins in the presence of DR+ or DQ+ cells, but only marginal levels of TNF production were induced in the presence of DP+ cells. Although both CD4+ T-cells and CD8+ T-cells produced TNF-alpha and TNF-beta in response to toxin stimulation in the presence of HLA class II+ cells, the former T-cell subset was the major source of producers of TNF-alpha and TNF-beta. PMID- 8707450 TI - The pro-oxidative riminophenazine B669 neutralizes the inhibitory effects of Mycobacterium tuberculosis on phagocyte antimicrobial activity. AB - The effects of clofazimine, a riminophenazine antimicrobial agent, and its analogue B669 on phagocyte functions have been investigated. Clofazimine, at concentrations attainable in vivo, and B669, in particular, increased the intracellular killing ability of phagocytes following appropriate cell stimulation. Similarly, nitro blue tetrazolium reduction, hydrogen peroxide production, lysosyme release and hexose monophosphate shunt activity were all increased by treating phagocytes with the riminophenazines. It has previously been shown that a 25 kDa glycolipoprotein derived from Mycobacterium tuberculosis inhibits phagocyte functions associated with phagocyte antimicrobial activity. The present study confirms these observations. A further aspect of the study examined the ability of riminophenazines to reverse the inhibition of phagocyte functions by the 25 kDa mycobacterial fraction. Whilst both riminophenazines were capable of partially but significantly reversing the inhibition due to the mycobacterial fraction, the restorative capacity of B669 was greater than that of clofazimine. PMID- 8707451 TI - The health care system, its doctors, patients, and illnesses. PMID- 8707452 TI - Self-reported depressive symptoms following treatment with corticosteroids and sedative-hypnotics. AB - OBJECTIVE: To evaluate associations between exposure to corticosteroids or sedative-hypnotic medications and incident self-reported depressive symptoms in medical inpatients. METHOD: The study utilized a prospective cohort design, focusing on acute depressive symptoms developing soon after medication exposure. The incidence of self-reported depressive symptoms was evaluated using a modified version of the Center for Epidemiological Studies Depression Rating Scale (CES D). The incidence of depressive symptoms in subjects newly exposed to corticosteroids and sedative-hypnotics was compared to that of a nonexposed comparison cohort. RESULTS: The incidence of self-reported depressive symptoms was elevated in subjects newly exposed to corticosteroids (Risk Ratio = 3.10), although the association did not attain statistical significance (p = .07). The risk ratio for sedative-hypnotic exposure was 4.18, a statistically significant finding (p = .02). As expected, incident self-reported depressive symptoms were also associated with several psychosocial variables. However, the data did not suggest that the observed associations between drug exposures and depressive symptoms were due to confounding by psychosocial or illness-related variables. PMID- 8707453 TI - Nonsteroidal anti-inflammatory drugs and severe psychiatric side effects. AB - OBJECTIVE: Nonsteroidal anti-inflammatory drugs (NSAIDS) are used extensively in the treatment of pain. This study explored the possibility that psychiatric side effects may be both more frequent and more severe than thought previously. METHOD: Four psychiatric outpatients, three with affective disorders and one with schizophrenia, were treated with NSAIDS for a complaint of pain. The NSAIDs were withdrawn, then restarted for three patients, and then withdrawn again one or more times. The patients were evaluated while on and off NSAIDs. RESULTS: All four patients developed moderate to severe depression and one became severely paranoid while on NSAIDs initially. When the NSAID was withdrawn there was remission of the depressive symptoms and in one case the accompanying paranoia. The depressive symptoms were reproduced when the NSAID was restarted in five instances (involving only 3 of the patients) and remitted when the NSAID was discontinued. One of these three patients also became paranoid in two instances. The paranoia remitted when the NSAID was discontinued. CONCLUSIONS: These findings suggest that NSAIDs can induce or exacerbate reproducible symptoms (depression, paranoia) in patients with either affective disorder or schizophrenia. These adverse effects may be more severe and frequent than thought previously. NSAID-treated patients should be studied for NSAID-induced psychiatric side effects. PMID- 8707454 TI - A comparison of the psychosocial aspects of AIDS and cancer-related bereavement. AB - OBJECTIVE: This study compares the psychological symptoms and bereavement distress of individuals bereaved by AIDS with a group bereaved by a cancer death, and addresses the question of whether an AIDS death is associated with a higher rate of adverse psychosocial factors that may increase risk of psychological morbidity in the bereaved individuals. METHOD: AIDS (n = 28) and cancer (n = 30) bereaved individuals (all within 3 months of the bereavement) completed measures of psychological morbidity and measures addressing a range of other adverse factors, e.g., number of losses, levels of social support and stigma. RESULTS: The cancer and AIDS bereaved were essentially similar on all psychological symptom measures. The AIDS group reported lower levels of social support in response to the bereavement than cancer bereaved individuals; a greater number of bereavements, were more likely to conceal the cause of death from significant others including their own family and perceived, in some instances, a greater level of rejection from others. The AIDS group reported higher levels of social support from friends than from family. CONCLUSIONS: At three months following bereavement, AIDS and cancer bereaved were similar in levels of distress. While this may change with the progress of grief over time, it suggests essentially similar early bereavement responses. Those bereaved by AIDS reported a range of other adverse factors such as a greater number of losses, lower social support, stigma, and less open disclosure of the cause of death. PMID- 8707455 TI - Mania due to general medical conditions: frequency, treatment, and cost. AB - OBJECTIVE: Mania due to general medicine conditions may occur in patients in a variety of settings. METHODS: We reviewed the charts of patients admitted to an adult psychiatric service over a nine-year period (Jan. 1985 to Dec. 1993). Patients were diagnosed with Organic Affective Syndrome (ICD-9 code 293.83) in 241 episodes (N = 227 patients). There were forty-seven manic or mixed episodes in forty patients (0.72% of all admissions). RESULTS: When DSM-IV criteria for Mood Disorder due to a General Medical Condition manic or mixed type (MDGMC) was applied, we found twenty-five patients with twenty-seven episodes (N = 30 treatment trials). Irritable mood predominated in twenty-seven (90%) of the thirty trials. CONCLUSIONS: Treatment included anticonvulsants in 63 percent, neuroleptics 63 percent, and lithium 40 percent. Favorable responses to anticonvulsants were seen; however combination therapy was used more frequently. Further research in this area is needed. PMID- 8707456 TI - Group psychological therapy for cancer patients: a point of view, and discussion of the hierarchy of options. AB - OBJECTIVE: We present here a point of view about the logical role of group psychological therapies in the care of cancer patients, and propose a comprehensive, four-stage program that could be implemented at any large cancer treatment center. METHOD: Our proposals rest on a review of the literature, and on our own experience with various kinds of psychological help for cancer patients, some of which is described. RESULTS: It is clear that group psychological therapies improve the quality of life of many participating cancer patients, and there is preliminary evidence that it may prolong life in some cases. We describe five main kinds of adjuvant psychological therapy, arranging them on a hierarchy of increasingly active participation by the recipient, and noting the status of evidence for their efficacy. They are: providing information, emotional support, behavioral training in coping skills, psychotherapy (of various kinds), and more speculatively, spiritual/existential therapy. CONCLUSIONS: We propose that it is time to consider psychological therapy as an adjuvant in cancer management, analogous to adjuvant chemotherapy. The main indication for advocating group psychological therapy to patients should, we argue, be an expert assessment that the patient is likely to benefit, and should not (as is usual at present) be restricted to either 1) those patients with overt psychopathology, or 2) those few who specifically request it. PMID- 8707457 TI - A pilot study of a short cognitive-behavioral group treatment for female recurrent suicide attempters. AB - OBJECTIVE: To test the feasibility and effectiveness of a high-frequency short term group cognitive-behavioral therapy (CBT) for recurrent suicide attempters. METHOD: CBT consisting of eight weekly and two "booster" sessions was given to nine female out-patients who had attempted suicide at least twice, seven of whom had a personality disorder. RESULTS: Four patients dropped out during the treatment; either because they were "chronic repeaters" (i.e., had a history of more than 8 suicide attempts), or because the last suicide attempt was too long ago. Although the patients were generally positive about the content of the therapy, no effects on psychiatric symptomatology or repetition of suicidal behavior could be demonstrated. CONCLUSIONS: Based on this experience, and on the two controlled studies of CBT of recurrent suicide attempters published in the literature, it is tentatively concluded that it is difficult to organize a short term high-frequency group treatment for recurrent suicide attempters with personality disorders, and that CBT may delay repetition of suicidal behavior, but that "major repeaters" (i.e., who made 4 or more suicide attempts) with a borderline personality disorder tend to stay major repeaters. PMID- 8707458 TI - Splitting on a pediatric consult liaison service. AB - OBJECTIVE: Although the concepts of splitting and projective identification have been useful in explaining certain group phenomena on adult psychiatric and medical wards, their application to pediatric settings has not been addressed in the literature. The authors demonstrate that early identification, staff conferencing, and family/staff conferencing can diffuse these dynamics in an academic pediatric setting. METHOD: The existing literature on splitting and projective identification is reviewed. Case vignettes are then used to illustrate the manifestations of splitting and projective identification in a pediatric setting and to demonstrate intervention strategies modified for children and their families from the adult literature. RESULTS: Splitting and projective identification can be interrupted in pediatric settings with early identification, staff conferencing, and family/staff conferencing. The cooperation of pediatric clinicians is critical in the implementation of these intervention strategies. CONCLUSIONS: The development of liaison support groups for pediatric residents and interdisciplinary treatment teams will enlist their cooperation in identifying splitting early, and in employing staff conferencing and family/staff conferencing to diffuse this group dynamic which, if left unchecked, can disrupt professional relationships and compromise the treatment of pediatric patients. PMID- 8707459 TI - Low-molecular-mass endothelial cell-stimulating angiogenic factor in relation to capillary growth induced in rat skeletal muscle by low-frequency electrical stimulation. AB - The involvement of endothelial cell-stimulating angiogenic factor (ESAF) in capillary growth was studied in adult rat skeletal muscles (tibialis anterior, TA, and extensor digitorum longus, EDL) in which capillary growth was induced by chronic unilateral electrical stimulation (10 Hz, 8 h/day for 7 days), and in sham-operated and unoperated control muscles. ESAF was assayed by its ability to activate latent collagenase in units per hour per milligram protein. Anatomical capillary density (CD, number of capillaries per mm2) and capillary per fibre ratio (C/F) were estimated in frozen sections from the same muscles after staining for endothelial alkaline phosphatase. In control muscles, ESAF levels were inversely related to capillary supply, being highest (1.82 +/- 0.25 units) in the glycolytic cortex of TA (CD 273 +/- 18/mm2, C/F 1.26 +/- 0.07), lowest (1.04 +/- 0.02 units) in its oxidative highly capillarized core (CD 862 +/- 60/mm2, C/F 2.05 +/- 0.04), and intermediate in EDL (ESAF 1.59 +/- 0.37 units, CD 527 +/- 26/mm2, C/F 1.44 +/- 0.06). Neither capillary supply nor ESAF levels were affected by sham operation. However, chronic electrical stimulation increased capillary supply significantly in EDL (CD 61% greater than in controls, C/F 45% greater) and ESAF levels were elevated 3-fold to 4.77 +/- 0.74 units. In TA muscles, stimulation increased capillary supply specifically in the glycolytic cortex (C/F 2.51 +/- 0.09, p < 0.0001 vs. control) and ESAF levels were increased significantly in this region to 3.19 +/- 0.55 units (p < 0.05, vs. control). C/F ratio and ESAF in the oxidative core of TA (2.31 +/- 0.05 and 1.48 +/- 0.23 units, respectively) were not significantly different from control values. Thus, chronic electrical stimulation, which is known to increase both shear stress and wall tension in capillaries and induce angiogenesis, also increased ESAF activity. PMID- 8707461 TI - Laser Doppler perfusion imaging--a new technique for measuring breast skin blood flow. AB - Future detection of breast cancer will probably be based on tissue function such as altered vascularity and blood flow. Assessment of tumour blood flow and its pattern may be significant for prognostic prediction, and early evaluation of treatment response. An ideal method of assessment of perfusion would therefore measure a wide area of the breast without the necessity for surface contact. A possible method for achieving this ideal is the recent development of a laser Doppler imager (LDI) that creates an image of tissue perfusion. The method is based on the recording of Doppler shift caused by movements of red blood cells in the backscattered light of a laser beam that successively scans a certain tissue area. This method reflects the vascularity of overlaying skin. We have applied this technique to 101 patients, 47 normal, 25 benign, and 29 carcinoma. Our findings were correlated with breast imaging and tissue diagnosis. The mean blood flow in 'normal' patients was 299 +/- 155 and 311 +/- 157 flux (+/- SD) for right and left breast, in patients with benign breast change it was 482 +/- 209 flux and in patients with carcinoma it was 711 +/- 280 flux. Patients with benign breast change have higher skin blood flow than symptomatic normal patients but the highest levels have been recorded in patients with breast cancer. We believe that LDI may have an important role in improving our understanding of the physiology of the normal breast and may provide a practical method of monitoring breast skin blood flow changes in clinical situations. PMID- 8707460 TI - Attenuated microvascular perfusion and reactivity in cardiac transplant recipients treated with cyclosporine. AB - Orthotopic heart transplant recipients (TX pts) treated with cyclosporine develop postoperative hypertension and their functional capacity remains less than normal. Altered responsiveness to adrenegic stimuli and impaired release of endothelial-derived relaxing factor are proposed mechanisms of cyclosporine induced raised peripheral vascular resistance. We compared responses to vasoconstrictor tests that stimulate sympathetic neural outflow (Valasalva maneuver and cold pressor test) and a vasodilator test that is dependent on the presence of a functionally intact endothelium (postocclusive hyperemia) in 16 TX pts with age-matched healthy controls, applying laser Doppler perfusion measurements (LDPM). Mean time since transplantation was 4.5 years (1-10 years). All TX pts received the triple regimen of prednisone, azathioprine and cyclosporine. Fourteen were considered hypertensive. Basal LDPM at rest expressed in arbitrary flux units (AU), was significantly lower in the TX pts (15.9 AU) than the controls (21.5 AU; p < 0.01). The maximal flux changes in the vasoconstrictor and vasodilator responses were comparable. However, the TX pts recovered faster from these responses and flux values at mid-to-late phase were lower following peak hyperemia and higher at any point following a cold pressor test than in the controls. Furthermore, a correlation was found between flux levels 30 s after either stimulus (r = 0.56; p < 0.0009) and time to reach prestimulus baseline after either test (r = 0.55; p < 0.002). With indirect evidence of comparable microvascular architecture, our findings suggest endothelial dysfunction in TX pts with intact functional capacity of the sympathetic nervous system. PMID- 8707462 TI - Quantitative reflection spectrophotometry: spatial and temporal variation of Hb oxygenation in human skin. AB - Applying a fast scanning reflection spectrophotometer and multicomponent spectra analysis, oxygen saturation (SHb) and relative concentration (CHb) of hemoglobin in the skin were determined at eight skin sites in 11 healthy persons. SHb was significantly higher at the tip of the index finger and big toe (90 +/- 3.9 and 92 +/- 4.2%, respectively) compared with the forehead, volar forearm, back of hand, abdomen, calf and forefoot where mean values varied between 52 and 67% (p < 0.001). CHb also was higher at acral sites (big toe: 2.04 +/- 0.14 arbitrary units (AU); index finger: 2.13 +/- 0.19 AU) than at the other locations (p < 0.0001) where it was between 0.56 +/- 0.12 AU (abdomen) and 0.95 +/- 0.28 AU (forefoot). In the course of time, rhythmical oscillations of both parameters at a frequency of 3-5/min were seen in 68% of the measurements, predominantly at the six proximal sites. Heating the measuring site to 44 degrees C caused a biphasic increase of CHb and SHb which was significant at the proximal sites (p < 0.0001). SHb values came into the range of arterial blood. Temporal and spatial variation of both parameters decreased. Reflection spectrophotometry gives the possibility to directly assess dermal hemoglobin saturation, its physiological variability and reactions to provocation stimuli. Concentration and saturation of hemoglobin in dermal vessels appear definitely different at acral compared with proximal sites. PMID- 8707463 TI - Postural vasoconstrictor response in human heart failure. AB - In order to study whether posturally induced vasoconstriction is impaired in subjects with heart failure, laser Doppler fluximetry was used to measure blood flow in the cutaneous microvascular bed of the foot at rest and during passive lowering of the extremity below heart level, in subjects with idiopathic dilated cardiomyopathy and in healthy controls. Two sites were studied: the toe pulp where arteriovenous anastomoses are numerous and the dorsum of the foot where such anastomoses are absent. Despite demonstrating a marked reduction in cutaneous blood flow at rest at each site [dorsum 3.0 AU (1.8-4.5) [median (range)] in heart failure patients vs 4.5 AU (1.8-31.6) in controls; toe 8.7 AU (3.1-33.5) in heart failure vs. 44.7 AU (5.2-280.0) in controls, p < 0.01], the results suggest that in non-oedematous subjects with severe left ventricular dysfunction there is no major disturbance of the postural vasoconstrictor response, either at a site rich in highly innervated anastomoses [43.6% (14.5 89.4) vs. 43.7% (15.6-91.1)] or in a site with few such anastomoses [79.7% (39.6 92.3) vs 69.6% (10.1-94.9)]. PMID- 8707464 TI - [Health professionals and tobacco: progress in Spain]. PMID- 8707465 TI - [Differences in hospital utilization between the autonomous communities of Madrid and Catalonia]. AB - Regional differences in hospital utilization have been long recognized. The objective of this study is to find out differences in hospital utilization and structure between Madrid and Catalonia. We calculate the crude and adjusted discharge and bed days rates and average length of stay from 1987 through 1989 for all diagnoses, and for 30 selected diagnoses by province of residence in 1987. We obtained the located and allocated beds, and the distribution of discharges by payers and of beds by providers for both regions, Catalonia shows consistently higher discharge rates (40% of public beds and 3.70 used beds) and Madrid has longer hospital stays (70% of public beds and 3.98 used beds). This work allows to formulate two questions about how health services operate: 1) organizational aspects could affect medical behavior. 2) hospital bed days could depend more on discharge rates than on length of stay. PMID- 8707466 TI - [Evolution in consumption of and attitude towards tobacco towards physicians at the regional hospital "Reina Sofia," Cordoba]. AB - OBJECTIVES: to study the evolution of the smoking prevalence and the personal attitudes about smoking among doctors of a university hospital. MATERIAL & METHODS: cross-sectional study. A survey. Study population was all physicians working in "Reina Sofia" University Hospital of Cordoba (N=738). By systematic sampling we select a random sample of 250 especialist physicians. We designed a self-administered questionnaire with items measuring the prevalence of smoking, the brand they used, the degree of smoking dependence, and their opinion about the new anti-smoking law. We also study the measures for smoking cessation that doctors considered most useful. We compared our data with other similar study carried out in 1987. RESULTS: the prevalence of tobacco use was 42% (CI:95%: 35.5 48.9), below the figure we obtained in 1987 (56.3%; p < 0.001). Doctors between 36-45 years had the highest rates of tobacco consumption (p < 0.01). 86.9% of smokers thought that smoking was unhealthy and among these 83.3% accepted the risk derived of smoking. 86.7% of smokers used to smoke inside the hospital. 86.8% of doctors thought that the new public places restrictive smoking law must be implemented. Doctors smoke more often (68.5%) when they are on call and 26.4% of them recognize to smoke when they inform the patient's family. CONCLUSIONS: we highlight a reduction of the prevalence of smoking among this kind of physicians. We still consider important the high number of physicians which smoke inside the hospital, although this figure have significantly decreased. PMID- 8707467 TI - [Demand and HIV testing prescription in primary care. Registry of the network of sentinel physicians from Castilla y Leon in 1991, 1992, and 1993]. AB - OBJECTIVE: the objective of this descriptive study is to improve the knowledge about the use and demand of HIV test in general population in Primary Health Care, as well as to detect lacks in general medicine related to HIV/AIDS. METHOD: from 1991 to 1993, the Red de Medicos Centinelas de Castilla y Leon, a voluntary sample of general practitioners and pediatrics has collected HIV test data from a covered population representative of the general population. RESULTS: one of the most important findings is the ascending frequency in tests done: On 65,107 and 125 per 100,000 inhabitants in 1991, 1992 and 1993 respectively. Intravenous drug users are the more important group risk in the register, 24%, followed by pregnant women, 17%, and the multiple heterosexual contacts, 9%. Among all the tests, 5.2% were positive. CONCLUSIONS: the comparison with other countries shows differences in frequency, cause and results, whose most probable explanation could be related to the different Health Care System and practice in general medicine and the patterns of risk infection in different populations. PMID- 8707468 TI - [Proposal of incentives for a fee-for-service medical practice]. PMID- 8707469 TI - [Evolution of suicide mortality in the Valencia Country 1976-1990]. AB - OBJECTIVE: suicide mortality rates have increased over last years in European countries. The aim of this study is to describe the mortality time trends for suicide in the Pais Valenciano between 1976-90. METHODS: from vital statistics data trends for suicide (E950-959 9th ICD) rates and adjusted-rates were analysed. A Poisson regression model was fit to analyse trends. RESULTS: suicide rates have increased for both sexes during the period up to 13.8 per 100,000 in males and 4.6 per 100,000 in females, being male rates three times females rates. The most important changes have been observed in the strong increase of male rates for age 15-24 who increased three times their rates. In women the higher increase belongs to age older than 65 years. CONCLUSION: the mortality increase in young male and in older remarks the need of interventions in order to prevent this important Public Health problem. PMID- 8707470 TI - Distributed gamma band responses in the brain studied in cortex, reticular formation, hippocampus and cerebellum. AB - The transient evoked responses of auditory cortex, reticular formation, hippocampus and cerebellar cortex to auditory stimulation have been analysed in the gamma frequency band on cats with chronically implanted electrodes. We found gamma band transient responses consisting of wave packets with 3-4 oscillations in all of the studied brain structures in the first 100 ms of the poststimulus period. The responses were strongly time-locked to the stimulation time point. The observation that the gamma band responses exist simultaneously in various brain structures supports the tentative proposal of our group on the "diffuse gamma response system" of the brain, which seems to be an important, universal operator in brain function. Furthermore, it shows that in search for generalized approaches to brain phenomena it is important to analyse the simultaneous behavior of different brain structures. PMID- 8707471 TI - Learning disabilities and learned helplessness: a heuristic approach. AB - This study investigated whether students with learning disabilities exhibited learned helpless behavior at a greater rate than their normal achieving peers when confronted with reading failure. Forty-five third grade students from a suburban elementary schools were participants in the study. Thirty of the subjects were classified as having a learning disability (LD) and the remaining 15 subjects were from regular education (RE) classrooms. Fifteen of the students with LD were placed in the treatment group and the remaining fifteen were placed in the control group. All the regular education students were placed in the treatment group. After randomly assigning the students with LD into either a treatment (stressed) group or a control (nonstressed) group, the stressed students were administered a reading instrument in order to measure how they dealt with failure. A one-way ANCOVA was conducted to determine whether significant differences existed between the groups based on their posttest scores. The results indicate that stressed students with LD have a significantly more difficult time recovering from stress than their regular education peers. PMID- 8707472 TI - Rapidly progressive form of Creutzfeldt-Jakob disease without dementia: a case report. AB - We describe a case of a patient with Creutzfeldt-Jakob disease (CJD) characterized by a rapid clinical course lasting one and a half months, by: presence of focal dystonic movements at onset, absence of mental deterioration in the period preceding the impairment of consciousness, ataxia, myoclonus and periodic EEG abnormalities. The autopsy confirmed subacute spongiform encephalopathy, but no evident neuronal loss was observed. An acute clinical course of CJD may explain this latter histological finding which, in turn, probably provides an explanation for the absence of intellectual impairment. PMID- 8707473 TI - P300 component of event-related potentials in passive tasks. AB - Auditory P300 components were recorded in two kinds of passive tasks (a sequence task and a novel task) and were compared to that in a standard oddball task. The P300, the amplitude of which was largest at Pz, was elicited in the two passive tasks. The P300 amplitude in the two passive tasks was not significantly smaller than that in the oddball task. No significant correlation was found between either the P300 latency or the P300 amplitude in the oddball task and those in the two passive tasks. These results suggested that the psychological significance of the P300 component in the passive tasks is different from that in the oddball task; however, the P300 in passive tasks is a useful tool in studying information processing in severely mentally ill subjects. PMID- 8707474 TI - Slowed reaction time performance on a divided attention task in elderly with environmental chemical odor intolerance. AB - Previous research has suggested an association between the subjective report of illness from environmental chemical odors and poorer cognitive task performance in persons with industrial levels of xenobiotic exposures. The present study investigated baseline morning performance on a computerized divided attention task in active retired adults without occupational exposures or clinical disorders who nonetheless rated themselves currently high versus low in episodic illness from the odor of certain environmental chemicals. The chemically intolerant group showed slower reaction times in registering both centrally and peripherally placed stimuli, but no difference in making target tracking errors. Measures of negative affect did not account for these findings. Taken together with evidence for heightened neurobehavioral sensitization in this population, the data suggest disturbances in allocation of attention and related cognitive functions. PMID- 8707475 TI - Infusion of glial maturation factor-beta reduces behavioral deficits after caudate nucleus injury in rats. AB - Adult rats with bilateral thermal lesions of the caudate nuclei (CN) show severe learning and memory deficits. The present study was designed to test the effects of an astroglial stimulating growth factor in this behavioral model. Immediately after receiving lesions of the CN, experimental subjects received an injection of one of three doses of glial maturation factor-beta (GMF-beta) directly in the lesion site. All subjects were then tested for twenty days on an active avoidance spatial alternation task. The behavioral recovery of the three groups of experimental animals was compared to that of animals having received the same brain damage and administration of a control substance (lesion controls), and to that of animals receiving a sham operation and no treatment (shams). The beneficial effects of administration were evident in the group of experimental animals receiving the lowest dose of GMF-beta. The performance of animals in this group was indistinguishable from that of the shams, and was significantly better than that of the lesion controls. The results suggest a behavioral role of GMF beta which, in an in vitro system, is known to be a growth regulator of astroglial cells. PMID- 8707476 TI - Response to tacrine in patients with dementia of the Alzheimer's type: cerebral perfusion change is related to change in mental status. AB - We evaluated change in cerebral hypoperfusion by single photon emission computed tomography (SPECT) in 12 patients with a clinical diagnosis of dementia Alzheimer's type (DAT) before and after treatment with Tacrine. Based on clinical evaluation of the SPECT scans before and after treatment, two groups of patients were identified. One (N = 5) experienced a relative decrease in cerebral perfusion and a decrease in mini-mental status examination (MMSE) scores across the treatment interval and was labeled "non-responders". A second group consisting of 7 patients was judged as showing an increase in cerebral perfusion at follow-up compared to baseline and an increase in MMSE scores. These were termed "responders". The group difference in MMSE score change was significant (Fisher's exact test: p < 0.008). Sensitivity (0.85) and specificity (1.0) and Receiver Operating Characteristic values (discriminability or d' = 3.12 and bias or beta = 4.6) indicated that direction of change in cerebral perfusion was a reasonable indicator of direction of change in MMSE scores. These results suggest that SPECT determined changes in cerebral perfusion may be useful in identification of DAT patients who are more likely to respond to centrally active agents such as tacrine. PMID- 8707478 TI - Resolution of partial cataplexy in multiple sclerosis by treatment with weak electromagnetic fields. AB - Cataplexy, an ancillary symptom of narcolepsy, involves the sudden loss of muscle tone without altered consciousness usually brought on by sudden excitement or emotional influence and extreme exertions (Guilleminault et al., 1974; Parks et al., 1974; Guilleminault, 1976; Aldrich, 1992; 1993; Scrima, 1981; Baker, 1985). Attacks of generalized cataplexy produce complete atonic, areflexic partial or complete paralysis of striated muscles commonly involving the leg muscles resulting in collapse of the knees and falling while milder forms often termed partial cataplexy may manifest by sagging of the face, eyelid, or jaw, dysarthria, blurred vision, drooping of the head, weakness of an arm or leg, buckling at the knees, or simply a momentary sensation of weakness that is imperceptible to observers (Guilleminault, 1976; Aldrich, 1993). The duration of cataplexy is usually a few seconds, although severe episodes can last several minutes and rarely several hours or days in the case of "status cataplecticus" (Parkes et al., 1974; Guilleminault, 1976; Billiard & Cadilhac, 1985; Aldrich, 1992; 1993). This report concerns a 51 year old man with chronic progressive multiple sclerosis who exhibited daily episodes of partial cataplexy which resolved within 3 weeks after he received treatment with picotesla electromagnetic fields. PMID- 8707477 TI - Event-related potentials during an auditory discrimination with prepulse inhibition in patients with schizophrenia, obsessive-compulsive disorder and healthy subjects. AB - Prepulse inhibition (PPI) is a measure of the influence of a stimulus (S1) on the response elicited by a second stimulus (S2) occurring shortly afterwards. Most S1/S2 measures of gating have used behavioural startle and the P50 event-related potential (ERP) amplitudes to detect PPI in a simple paired stimulus paradigm. We report on two behavioural (reaction time, RT, and the electromyographically recorded response of the musculus orbicularis oculi, EMG) and 5 ERP measures of PPI where S2 was the target in an auditory two-tone discrimination. Subjects were 21 healthy controls (CON), 11 obsessive-compulsive (OCD) and 9 schizophrenic patients (SCH). The prepulse 100 ms before S2 induced more omission errors and longer RTs compared to 500ms S1-S2 interval in all subjects. PPI was also evident in EMG, P50, N1, P3 but not P2 or N2 amplitudes of CON subjects. SCH patients showed attenuation of PPI on the same measures. OCD patients were characterized only by their slow RT and a marginal attenuation of PPI of the EMG response. A correlational analysis implied separate relationships of ERP indices of PPI to the cognitive and psychomotor consequences of the prepulse on behavioural and discrimination responses. However, SCH patients showed a general rather than a specific impairment of these indices. PMID- 8707479 TI - Effect of weak electromagnetic fields on the amplitude of the pattern reversal VEP response in Parkinson's disease. AB - Visual evoked potential (VEP) studies are widely used for the diagnosis of multiple sclerosis (MS) and are also useful in monitoring the effects of various therapeutic modalities in the disease. Prolongation of the VEP latencies has been demonstrated in patients with MS and in other neurodegenerative disorders including Parkinson's disease (PD), a disorder characterized by deficient cerebral dopamine (DA) functions. Pharmacological and biochemical studies have demonstrated a positive correlation between the amplitude of the VEP response and cerebral DA levels. Since brief, extracerebral applications of picotesla (pT) range flux intensity electromagnetic fields (EMFs) of low frequency have been shown to produce rapid improvement in motor and cognitive symptoms in PD, it is expected that application these EMFs would lead also to an increase in the amplitude of VEP response. This report documents three randomly selected PD patients who, following two successive brief extracerebral applications of pT range EMFs, showed an almost 3-fold increase of the mean pretreatment amplitude of the pattern reversal VEP in response to monocular stimulation. One patient underwent also a placebo EMF treatment which did not result in a significant change in the posttreatment amplitude. The study demonstrates that in Parkinsonian patients extracerebral application of these EMFs rapidly increases in amplitude of the VEP response and, by inference, cerebral DA levels presumably by increasing DA release. PMID- 8707480 TI - Treatment with weak electromagnetic fields improves fatigue associated with multiple sclerosis. AB - It is estimated that 75-90% of patients with multiple sclerosis (MS) experience fatigue at some point during the course of the disease and that in about half of these patients, subjective fatigue is a primary complaint. In the majority of patients fatigue is present throughout the course of the day being most prominent in the mid to late afternoon. Sleepiness is not prominent, but patients report that rest may attenuate fatigability. The pathophysiology of the fatigue of MS remains unknown. Delayed impulse conduction in demyelinated zones may render transmission in the brainstem reticular formation less effective. In addition, the observation that rest may restore energy and that administration of pemoline and amantadine, which increase the synthesis and release of monoamines, often improve the fatigue of MS suggest that depletion of neurotransmitter stores in damaged neurons may contribute significantly to the development of fatigue in these patients. The present report concerns three MS patients who experienced over several years continuous and debilitating fatigue throughout the course of the day. Fatigue was exacerbated by increased physical activity and was not improved by rest. After receiving a course of treatments with picotesla flux electromagnetic fields (EMFs), which were applied extracranially, all patients experienced improvement in fatigue. Remarkably, patients noted that several months after initiation of treatment with EMFs they were able to recover, after a short period of rest, from fatigue which followed increased physical activity. These observations suggest that replenishment of monoamine stores in neurons damaged by demyelination in the brainstem reticular formation by periodic applications of picotesla flux intensity EMFs may lead to more effective impulse conduction and thus to improvement in fatigue including rapid recovery of fatigue after rest. PMID- 8707481 TI - Effects of picotesla flux electromagnetic fields on dopaminergic transmission in Tourette's syndrome. AB - Tourette's syndrome (TS), a chronic familial neuropsychiatric disorder of unknown etiology, is characterized clinically by the presence of motor and vocal tics that wax and wane in severity over the time and by the occurrence of a variety of neurobehavioral disorders. It is believed that the tics of TS result from increased dopamine (DA) activity caused by postsynaptic DA receptor supersensitivity. The synthesis and release of DA is regulated presynaptically by a specific class of DA D2 receptors, termed autoreceptors activation of which causes inhibition of DA synthesis and release. In experimental animals and humans administration of small doses of apomorphine, a DA D2 autoreceptor agonist, produces yawning. Recurrent episodes of yawning followed by increased motor tic activity was observed in two patients with TS during exposure to brief, extracranial applications of picotesla flux electromagnetic fields (EMFs). On the basis of these observations it is suggested that recurrent episodes of yawning in response to application of EMFs was induced by activation of presynaptic DA D2 autoreceptors while further exposure to these EMFs caused excessive stimulation of postsynaptic DA D2 receptors resulting in exacerbation of the tics. Thus, the dual effects of picotesla flux EMFs on the DA D2 autoreceptor and the postsynaptic receptor resemble the biphasic pharmacological and behavioral properties of apomorphine, a DA agonist which activates the autoreceptors in low doses while in higher doses causes stimulation of the postsynaptic receptors producing exacerbation of symptoms of TS. These findings demonstrate that picotesla flux EMFs applied extracerebrally may influence nigrostriatal DA transmission at pre- and postsynaptic DA D2 receptor sites. PMID- 8707482 TI - Involvement of delta-opioid receptors in immunosuppression. AB - The present experiments demonstrate that: (1) activation of delta-opioid receptors by a highly selective agonist DSLET leads to immunosuppression, whereas the blockade of these receptors by enkephalin analog ICI 174864 produces immunostimulation; (2) the immunomodulating effect of delta-opioid receptors is mediated by central mechanisms involving the hypothalamus-hypophysis complex; (3) the postsynaptic 5-HT (serotonin) receptors of 5-HT-2 type are involved in the immunoinhibitory action of DSLET. On the other hand, the immunostimulating effect of ICI 174864 is realized with the participation of D-2 postsynaptic DA (dopamine) receptors. It has also been shown that thymus is implicated in the immunostimulating influence of ICI 174864. The data obtained are explained as a result of the interaction of the neuromediator and opioid peptidergic systems in neuroimmunomodulation. PMID- 8707484 TI - Psychogenic stress induces chromosomal and DNA damage. AB - In this investigation, rats subjected to swim stress showed within 24 hours significant increases in both the level of chromosome aberrations and Sister Chromatid Exchanges (SCEs) in bone marrow cells. The generality of cytogenetic damage by behavioral stressors was demonstrated by exposing rats to both cold-and warm-water forced swims, to white noise, and to continuous or intermittent inescapable foot shock stress (IFS). The induction of chromosome aberrations and SCEs, to differing degrees, by stressors that differ both quantitatively and qualitatively, demonstrates that this is a general phenomenon of stress. The use of an additional measure, unscheduled DNA Synthesis (UDS) showed that stress induced genotoxic damage can occur in a second cell type and on a molecular as well as chromosomal level. These results indicate that there may be a cellular genetic basis for some of the effects of stress. PMID- 8707483 TI - Massage therapy is associated with enhancement of the immune system's cytotoxic capacity. AB - Twenty-nine gay men (20 HIV+, 9 HIV-) received daily massages for one month. A subset of 11 of the HIV+ subjects served as a within subject control group (one month with and without massages). Major immune findings for the effects of the month of massage included a significant increase in Natural Killer Cell number, Natural Killer Cell Cytotoxicity, soluble CD8, and the cytotoxic subset of CD8 cells. There were no changes in HIV disease progression markers (CD4, CD4/CD8 ratio, Beta-2 microglobulin, neopterin). Major neuroendocrine findings, measured via 24 hour urines included a significant decrease in cortisol, and nonsignificant trends toward decrease of catecholamines. There were also significant decreases in anxiety and increases in relaxation which were significantly correlated with increases in NK cell number. Thus, there appears to be an increase in cytotoxic capacity associated with massage. Implications for HIV+ men as those with other illnesses, particularly cancer, are discussed. PMID- 8707485 TI - The use of conditioning to probe for CNS pathways that regulate fever and NK cell activity. AB - Immune and central nervous system (CNS) interactions are complicated because afferent signals from the immune system to the CNS in response to antigens or infections may elicit an immediate efferent response to the immune system. This communication loop is required for the homeostatic regulation of the immune system. Conditioning can be used as a tool to take the communication loop apart. In conditioned animals, the conditioned stimulus can be employed later to trigger the site of the association memory located within CNS, and set off the efferent pathway. Conditioning therefore allows one to isolate and identify the potential circuits in the brain that becomes conditioned. We have conditioned a pathway in the brain which can be used to modulate core body temperature (Tc) and natural killer (NK) cell activity. The Tc and NK cell activity are used as readouts to detect the expression of the conditioned response which is taking place in the brain. Since various cytokines (IFN, IL-1 etc) that are produced by antigenic stimulation invariably raise fever, it appears that the immune system could signal the CNS with nonspecific cytokines that activate the hypothalamic pituitary pathway to modulate core body temperature. These observations infer that the thermoregulatory pathway in the brain becomes conditioned and points to a common pathway of communication in which interferon-beta, prostaglandin E2, CRH and ACTH appear to play a role in modulating both Tc and NK cell activity. PMID- 8707486 TI - Brainstem auditory evoked potentials in respiratory insufficiency following encephalitis. AB - Brainstem auditory evoked potentials (BAEPs) were recorded in 14 artificially ventilated patients (12 males, 2 females; mean age 33.3 +/- 16.3 years, range 18 67) with respiratory insufficiency resulting from severe inflammatory encephalopathies. The results were compared with those of 17 healthy volunteers (13 males, 4 females; mean age 27.4 +/- 5.3 years, range 21-45). BAEPs in the study patients showed prolonged interpeak latencies (I-III, I-V, III-V, IV-V) and delayed absolute latencies of waves I, II, III, and V at least on one side. Because the auditory pathways are in the near vicinity of the respiratory control centers in the brainstem, the electrophysiologic abnormalities of wave III and the IV/V complex may be a reflection of the disturbed central control of ventilation. PMID- 8707487 TI - Transcranial Doppler assessment of cerebral flow velocity at rest and during voluntary movements in young and elderly healthy subjects. AB - Fifty-five healthy volunteers (thirty young and twenty-five elderly), all of them right-handed, were investigated by Transcranial Doppler monitoring mean flow velocity on the right and left middle cerebral artery at rest, during a motor task performed by the hand and in the following rest condition. In all cases the motor task induced an increase in bilateral mean flow velocity, but significantly greater on the opposite side middle cerebral artery in the task. The percentage increase in mean flow velocity compared to the rest condition observed during the motor task performed by the opposite side hand to the artery investigated was significantly greater in the young than the elderly. Moreover, in the young, this increase showed a slight prevalence during the left middle cerebral artery assessment relating to right hand preference. Furthermore, elderly subjects achieved the greatest mean flow velocity percentage increase compared to rest condition later than young subjects and returned to basal values more slowly. PMID- 8707488 TI - Underlying factor structure of perinatal events. AB - The underlying factor structure of perinatal events was examined for 847 children (272 normals, 117 developmentally disabled, 37 speech disordered, 221 learning disordered, 91 emotionally disturbed, and 109 Head Start children). Pertinent perinatal information from the Maternal Perinatal Scale was subjected to a principle-components factor analysis. Ten factors accounting for some 58% of the total variance were isolated. In general, these factors appeared to be consistent with clinical evidence. The results were discussed in terms of their implications for the utility of perinatal information. PMID- 8707489 TI - Auditory affect perception in a dichotic listening paradigm as a function of verbal fluency classification. AB - This study evaluated cerebral asymmetry for affect perception as a function of fluency classification. After being assigned to a fluency category using scores on the FAS test (Borkowski, Benton, & Spreen, 1967), forty-five right-handed subjects with normal auditory acuity listened to the Bryden and MacCrae (1989) Dichotic Emotional Words Tape. Subjects higher in fluency exhibited significantly greater right and left ear advantages than subjects lower in fluency. Conversely, REA scores for words were significantly greater than REA scores for affect, while LEA scores for affect were significantly greater than LEA scores for words. PMID- 8707490 TI - Differential visuospatial dysfunction following striatal, frontal white matter, or posterior thalamic infarction. AB - The present study investigated visuospatial functioning in 35 right-hand dominant patients with striatal, frontal white matter, or posterior thalamic infarction, in the right cerebral hemisphere. On tasks of visual detection and discrimination, the groups performed within normal limits and comparable to one another. On tasks of visual synthesis and spatial analysis, each group was significantly impaired, with greatest impairment following infarction of deep grey structures. Correlational analyses indicated that in the striatal and frontal white matter groups, visual synthesis and spatial analysis task performance were strongly correlated with performance on neuropsychological tests of executive control. This relationship was not observed in the posterior thalamic group. The results support the hypothesis of differential patterns of visuospatial performance depending on the anatomical locus of subcortical lesions. The findings are explained in terms of differing subcortical circuitry and suggest that multiple substrates mediate the sequelae of subcortical lesions. PMID- 8707491 TI - Emotion-specific processing deficit in focal brain-damaged patients. AB - Focal brain-damaged patients (left hemisphere damage, right hemisphere damage) and hospitalized general medical patients were asked to sort test photographs into target expressions of four facial emotions, happy, sad, fear and anger. In a second task, patients were asked to match neutral photographs with these target emotion expressions in a forced-choice format. Patients were also asked to rate their mood state on a two-dimensional affect grid. Right hemisphere-damaged patients were significantly inaccurate in matching the test-target expressions of facial emotions in comparison to left hemisphere-damaged or general medical patients. Analysis of error scores indicated a bias toward negative emotions by left hemisphere-damaged patients. Congruent to their mood state, left hemisphere damaged patients also attributed "sadness" on neutral state of expression significantly more often than in right hemisphere-damaged patients. PMID- 8707492 TI - Concurrent and predictive validity of an early childhood screening test. AB - The purpose of this study was to validate an early childhood screening instrument concurrently with other established psychoeducational tests and predictively with a standardized measure of achievement. A battery of tests including a screener and five other established psychoeducational measures was administered to 64 children during their kindergarten year. An achievement test was administered to 33 of these children during their third grade year. The screening test scores were found to be significantly related to the results of other psychoeducational measures both concurrently and predictively. The instrument permits teachers to gather behavior samples in minutes. PMID- 8707493 TI - Introduction: the quality of the medical evidence: is it good enough? PMID- 8707494 TI - Generations of misplaced confidence. A personal view. AB - A humorous look at one family's experience with the milestones of medical achievement illustrates the need for continued research on the efficacy of treatment. Skepticism about the benefits of new treatments and careful examination of even standard procedures should be encouraged within the medical profession. PMID- 8707495 TI - Assessing the quality of randomized controlled trials. Current issues and future directions. AB - Assessing the quality of randomized controlled trials is a relatively new and important development. Three approaches have been developed: component, checklist, and scale assessment. Component approaches evaluate selected aspects of trials, such as masking. Checklists and scales involve lists of items thought to be integral to study quality. Scales, unlike the other methods, provide a summary numeric score of quality, which can be formally incorporated into a systematic review. Most scales to date have not been developed with sufficient rigor, however. Empirical evidence indicates that differences in scale development can lead to important differences in quality assessment. Several methods for including quality scores in systematic reviews have been proposed, but since little empirical evidence supports any given method, results must be interpreted cautiously. Future efforts may be best focused on gathering more empirical evidence to identify trial characteristics directly related to bias in the estimates of intervention effects and on improving the way in which trials are reported. PMID- 8707496 TI - Influences on the quality of published drug studies. AB - To practice evidence-based medicine, physicians need data on the clinical effectiveness, toxicity, convenience, and cost of new drugs compared with available alternatives. We give examples of published drug studies that are defective, sometimes because pharmaceutical industry funding has affected their content and quality. We make recommendations on how to avoid these defects. PMID- 8707497 TI - Beware of surrogate outcome measures. AB - Surrogate outcome measures may speed up clinical research if they can be measured earlier in a study than the primary outcome of interest. We review the justification for their use and conclude that reliance on them may be harmful. Results obtained with surrogates should therefore be regarded as preliminary. Large, definitive trials with clinically relevant outcomes should always be performed before new interventions are accepted. PMID- 8707498 TI - Competing or complementary? Ethical considerations and the quality of randomized trials. AB - The ethical principles of beneficence (doing the best for one's patients and clients), nonmaleficence (doing no harm), autonomy, justice, and equity, are positively associated with the quality of a trial rather than being opposed to it. Ethical considerations involve such important questions as appropriate design and documentation, participation rates, withdrawals or losses to follow-up, careful monitoring of safety and efficacy, timely completion, generalizability, and publication and dissemination of findings. When conflicts between ethical considerations and the trial's scientific quality do occur, the ethical considerations must be paramount. PMID- 8707499 TI - Bias in the analysis and reporting of randomized controlled trials. AB - The most reliable information on any type of medical intervention is provided by the results of randomized clinical trials (RCTs). In response to increasing pressure to make effective use of limited resources, increasing numbers of health professionals rely on the medical literature, in particular reports of RCTs. However, RCTs may be influenced by a number of factors that introduce bias during the conduct, analysis, and reporting of the trial. Trials may be described as random, when in fact only quasi-random means of treatment allocation have been used; patients may be selectively removed from the analysis; and the report may restrict presentation to or give undue emphasis to only the analyses that yield positive results. The implications of such bias are discussed with particular reference to the effect that they may have on reviews and meta-analyses. PMID- 8707500 TI - Collecting the evidence systematically. Ensuring that it is complete and up-to date. AB - A complete and comprehensive search of the relevant evidence is an essential step in the preparation of a scientific systematic review. The Cochrane Pregnancy and Childbirth Database consists of some 600 systematic reviews, based on almost 6,000 randomized or quasi-randomized trials of care during pregnancy and childbirth. As ascertainment from electronic search of the National Library of Medicine MEDLINE database was not complete, the mainstay of our search strategy was a systematic hand search of some 60 journals, beginning with volumes published in 1950. Additional references were obtained from the list of references in primary research reports and conference reports. A major effort was made to identify unpublished trials and to obtain unpublished data from published trials. PMID- 8707501 TI - Using the evidence. A need for quantity, not quality? AB - The use of research evidence depends on more than its quality. If one considers other factors that influence the use of evidence, the quantity of accessible evidence and initiatives ensuring a wider understanding of it must be equally important. In consequence, improving the quality of research evidence alone will not necessarily lead to its greater use. PMID- 8707502 TI - The 'utility' of the visual analog scale in medical decision making and technology assessment. Is it an alternative to the time trade-off? AB - Methods often used for the valuation of health states are the time trade-off (TTO) and the visual analog scale (VAS). The VAS is easier than the TTO and can be self-administered; however it usually leads to lower scores. In the literature a power transformation of group mean VAS scores to TTO scores has been proposed. We were able to replicate this finding of a power function. We found coefficients that were very similar to those from the literature, for 183 cancer patients. The relationship existed independently of disease state and health status. PMID- 8707503 TI - Transcutaneous electrical nerve stimulation (TENS): a technology assessment. AB - The scientific evidence for clinical effectiveness of transcutaneous electrical nerve stimulation (TENS) for treatment of acute, chronic, and labor and delivery pain is assessed in this paper, and it is concluded that there is little evidence for other than a limited use of TENS. The utilization of TENS in Canadian hospitals and payments for TENS services are addressed. Some practicalities regarding the use and assessment of health technologies are discussed. PMID- 8707504 TI - Hospital- and patient-related characteristics determining length of hospital stay for hip and knee replacements. AB - Total hip arthroplasty (THA) and total knee arthroplasty (TKA) are among the most prestigious health care technologies. Their popularity has grown rapidly, and an increasing proportion of health care resources is allocated to them. We studied patient- and hospital-related factors that cause variation in a major determinant of hospital costs, the length of hospital stay (LOS) for THA and TKA. We gathered data on 10,288 hip and 5,173 knee patients with primary or secondary arthrosis from the Finnish Arthroplasty Register, which we linked with the Finnish Hospital Discharge Register. Patient- and hospital-related variations in LOS were explained using regression models. Of the patient-related factors, complications caused the greatest prolongation of hospital stay, but patient's age, gender, and charge category also influenced LOS. Hospital-related factors were major causes of LOS variation. In the hospitals the average case-mix-adjusted LOS ranged from less than a week to 3 weeks. The number of arthroplasties performed in hospital was inversely related to LOS. The within-hospital LOS figures for THA and TKA were strikingly similar and persistent. PMID- 8707505 TI - Economic evaluation of HIV testing among intravenous drug users. An analytic framework and its application to Italy. AB - We performed an economic evaluation of HIV testing among intravenous drug users (IVDUs) in Italy using the analytical framework of cost-effectiveness analysis. A semi-Markov model was developed to calculate costs and life expectancy of a cohort of IVDUs with and without an annual HIV testing program. We also investigated the incremental cost-effectiveness of a hypothetical early treatment to prolong the life expectancy of HIV-infected asymptomatic subjects by 1 year. The testing program is cost saving in the low prevalence scenario (0.05), and costs L 14,000,000 (US $8,400) and L 55,800,000 (US $33,500) per year of life saved in areas of medium (0.3) and high (0.6) prevalence, respectively. The incremental cost-effectiveness of the hypothetical early treatment may compare favorably with other health care interventions. The program may be considered a cost-effective procedure in low and medium prevalence areas. Where prevalence is high, more evidence about the magnitude of the behavior change is needed. In these areas, the availability of an effective early treatment may become the economic rationale for implementing such a program. PMID- 8707506 TI - The impact of the technological care environment on the nursing role. AB - Proliferation of acute health care technology creates problems and benefits for nurses and patients. In this paper the impact of technology on the nursing work role is reviewed through the international literature. The thrust of the nursing literature has, not surprisingly, matured over time as the use of technology has become well established in the acute care environment, and three themes can be identified. The implications for acute care nurse specialists, including their educational needs, are set in context of the Australian health care system, with particular reference to the cardiac care environments. PMID- 8707507 TI - Awareness of providers' use of new medical technology among private health care plans in the United States. AB - In a national survey of medical directors at 231 U.S. private health care plans that cover over two thirds of the privately insured population, we studied whether medical directors are aware when a new technology, such as laser therapy, is being used in procedures for which claims are submitted, the factors alerting them to such use, and the factors prompting them to make a specific coverage decision for the technology. We also examined possible associations between health plans' characteristics (HMO versus indemnity, size, profit status, and time in operation) and their medical directors' awareness of the use of technologies, factors alerting medical directors to their use, and factors prompting specific coverage decisions. The majority of plans were generally not aware that laser technology was being used when it was billed under a general billing code, raising the possibility that less effective or less safe technologies could be introduced rapidly into the treatment of insured populations. Nonprofit and older plans were less likely to be aware that lasers were used in some procedures than for-profit and younger plans. PMID- 8707508 TI - Abdominal and vaginal radical hysterectomy among U.S. women aged 65 years and older. AB - Unlike most European and Asian countries, radical vaginal hysterectomy (RVH) is not performed often in the United States, especially among older women. To examine the changes in RVH over the years, trends in hospital stay, hospital charges, and patient survival, we studied women aged 65 years and older undergoing RVH and compared them with patients receiving radical abdominal hysterectomy (RAH). During the study period there were a total of 288 RVH surgeries compared with 4,835 RAH surgeries. There were no significant changes in the number or proportion of RVH patients over 8 years (p = .50, trend test). On the average, RVH patients were significantly older and had shorter hospital stays. Among patients without cancer, there were no significant differences in the age, race, or survival of patients having either RVH or RAH. Similar results were obtained for patients with cancer. PMID- 8707509 TI - Evaluation of the regular practice of breast cancer screening in a health area. AB - Breast cancer screening has proved to be efficacious only in clinical trials or structured programs. However, little is known of its effectiveness as a regular practice. This study is an assessment of breast cancer screening in regular practice in a Spanish area of 350,000 inhabitants through analysis of existing sources of information. It is evident that this screening is insufficient, inefficient, very variable, nonspecific, and probably ineffective, although there is insufficient information to prove it. The immediate implementation of a well structured and accredited screening program is essential to overcome the current limitations. PMID- 8707510 TI - Priority classification of patients according to a modified 'Norwegian model'. AB - A priority classification was evaluated according to a modified 'Norwegian model.' Many diseases do not belong to any specific priority category based only on the diagnosis. The classification also depends on the condition's type, site, and phase as well as the patient's age and overall condition. Savings cannot be achieved by the model used because 89% of the patients belonged to the priority categories I-III, the care of which can be classified as necessary. PMID- 8707511 TI - Impact of health reorganization on nurses in New Zealand. AB - New Zealand, like many other Western countries, has undergone widesweeping restructuring in its public sector over the past few years. Embracing a market economy, successive governments--both liberal and conservative--have encouraged competition and deregulation. Below, how the New Zealand nurses reorganized to meet this new challenge. PMID- 8707512 TI - The future of the clinical nurse specialist in the USA. AB - In light of the climate of healthcare reform, the future of the traditional clinical nurse specialist is tenuous while that of the nurse practitioner is bright. The merger of these two traditional roles into a unified advanced practice nurse (APN) role is critical now for the profession of nursing, for patients and for the entire healthcare arena. The following describes how the APN will be a leader in providing health care both in the USA and internationally and a primary mover toward WHO's goal of "health for all". PMID- 8707513 TI - The impact of multiskilling on quality of care. AB - Hospital finances in the USA are in a crisis. While profits earned from the healthcare industry appear to be at an all-time high, individual hospitals are going bankrupt. Hospitals are not generating sufficient income to pay their expenses given how they have traditionally operated. Hence, we are witnessing the merger and consolidation of competitors, resulting in significantly fewer hospital beds in certain geographical areas. In addition to the strategy of closing hospital beds, severe budget restrictions are impacting upon all remaining services within the hospitals. PMID- 8707514 TI - Odyssey of a project to improve psychogeriatric nursing care. AB - A grant to prepare a tool to evaluate quality of care in Israeli psychogeriatric units developed into a longitudinal project to improve care. Below, how it began as an "idea" with defined boundaries and grew into an open-ended dynamic "ideology." PMID- 8707515 TI - What restructuring means for nurses in Denmark. AB - Irrespective of structural and other changes, today the healthcare sectors in all countries share many common problems, particularly in regard to: whether to allow private financing via insurance or public financing via taxation; higher consumer demand and expectations; greater opportunities for medical science interventions; and prioritizing treatment and patient categories. Below, how the Danish healthcare sector is facing these challenges and what it means for nurses. PMID- 8707516 TI - Preliminary report from the Irish Intellectual Disability Database. AB - The Department of Health established a computerised national mental handicap database in the Republic of Ireland in 1995 for the purposes of providing accurate information for planning services for people with mental handicap. Initial estimates indicate that 27,193 people are either in receipt of or in need of mental handicap services. This paper presents preliminary prevalence data from the database which were analysed by age, sex, degree of handicap and geographical region. Comparisons are drawn with the findings of two previous censuses which are conducted in Ireland in 1974 and 1981 and implications for future service provision are discussed. PMID- 8707517 TI - A memory clinic for the assessment of mild dementia. AB - The aims and experience of work in a specialist memory clinic' are described with reference to the first 200 patients attending the service. 93.5% of attenders were found to suffer from dementia. 67.9% of these fulfilled criteria for probable/possible Alzheimer's disease or other primary degenerative dementias and 8.6% fulfilled criteria for vascular dementia. In the remaining cases the aetiology of the dementia was considered to be multifactorial. The memory clinic proved to be an important resource for demented patients, their carers, and for research projects. PMID- 8707518 TI - Cyclospora species--a cause of diarrhoea among Irish travellers to Asia. AB - We report three cases of travellers' diarrhoea caused by Cyclospora cayetanensis. These demonstrate the importance of cyclosporiasis as a cause of diarrhoea in Irish travellers to Nepal and Pakistan and its effective treatment with co trimoxazole. Laboratory staff must measure the size of oocysts in stool samples to properly identify this protozoan. PMID- 8707519 TI - An analysis of the effects of HIV infection in a cohort of intravenous drug users. AB - Intravenous drug use has been a serious problem for Dublin's deprived inner city areas since the late 1970's. In 1985, a cohort of all known intravenous drug users (IVDUs) was identified in one inner city electoral cohort and has been followed since then. The cohort was identified before the introduction of HIV testing. The prevalence of HIV infection and its consequences have continued to rise. By 1994, 80% of the group were known to have had a HIV test: 53 (65%) are seropositive. Twenty-one (26%) have died, almost all from the seropositive group. Thirty-six (44%) continued to inject heroin during 1994. The implications of these grim statistics are discussed. PMID- 8707520 TI - The effects of hospital admission on drug prescribing in an elderly population. AB - 307 consecutive patients admitted to our unit were studied. The reasons for altering patients' drug regimens were noted. The price difference between the patients' drugs on admission to those on discharge was calculated. While over 40% of drugs regimens on admission were unchanged on discharge some adjustments were made in the remainder. The most common reason for stopping a drug was that the hospital physician could find no indication for its use. A 30.3% reduction in price of discharge drugs compared with drugs on admission was achieved. PMID- 8707521 TI - Medical emergency--Rwanda 1995. A personal experience. PMID- 8707522 TI - Withdrawal of routine psychiatric medications in a general medical ward. PMID- 8707523 TI - Babies born before arrival. PMID- 8707524 TI - Stammering still prevalent in Ireland. PMID- 8707526 TI - Lactose intolerance. PMID- 8707525 TI - Storage and handling of vaccines by family doctors. PMID- 8707527 TI - Medical treatment as an alternative to surgery in the management of benign prostatic hypertrophy. PMID- 8707528 TI - Idiopathic constipation. PMID- 8707529 TI - Bone density profiles and osteoporosis incidence in Irish females. AB - 1725 Irish females aged 15-70+ were included in this analysis to establish the group bone mineral density (BMD) profiles. Each subject was screened for 27 risk factors by questionnaire. Subjects were segregated into normal (N) and non-normal (NN) subgroups on the basis of the questionnaire. BMD of lumbar spine and hip were measured by dual energy X ray absorptiometry (DEXA). A substantial difference in BMD at all sites was observed between subgroups even following adjustment for body mass index and years of exposure to oestrogen. In the subgroup designated as non-normal 42%, 54%, 72% and 69% of those studied had spinal BMD levels > 2SD below mean peak at 55-59, 60-64, 65-69 and 70+ respectively. In comparison, similar age cohorts in the subgroup designated normal were 26%, 46%, 44% and 68% respectively. A marked difference was also observed in the younger age groups (15-34) where the percentage of individuals with spinal BMD levels > 2SD below mean peak was < 1.9% for N and 18.8% for NN. The results of this study suggest that the use of a questionnaire may be a valuable low cost screening tool in the younger groups. PMID- 8707530 TI - European Nosocomial Infection Survey: analysis of Irish data. Irish Intensive Care Nosocomial Pneumonia Survey Group. AB - The European Nosocomial Infection Survey enrolled 9127 patients in 14 countries providing epidemiological data on pneumonia in 325 Intensive Care Units (ICU). Ten ICUs took part in Ireland recruiting 205 patients. Using strict diagnostic criteria, the pneumonia rate for the Irish group was 11.7%; European-wide it was of 11.8%. The ICU mortality Ireland was 14.2%, in the overall survey it was 14.3%. The pneumonia mortality rate in the overall survey was 30%. PMID- 8707531 TI - Suicide levels in Cork City 1987/1991. AB - This study investigates the level of suicide in Cork City during the 5-year period 1987/1991. Both male and female crude rates were higher than the corresponding national levels. The male/female ratio was 2.3:1, which is consistent with established findings. Vulnerability to suicide is most pronounced in 20-39 year old males and females in the 40-59 age category. There was some evidence of an increased risk of suicide among single elderly males; otherwise marital status was not a significant factor. A positive association was noted between unemployment and suicide rate for males. Drowning was used as the method of suicide by approximately half the male and female sub-groups; this was followed by hanging in the case of males and overdosing among females. However all suicide cases under 20 years of age used hanging. PMID- 8707532 TI - Relationships of nurses' critical thinking ability and perceived patient self disclosure to accuracy in assessment of depression. AB - This descriptive study examined the relationships of nurses' critical thinking ability and perceived patient self-disclosure to accuracy in nursing assessment of depression in older medical patients. The sample consisted of 120 female nurse patient dyads. Critical thinking ability was measured by the Watson-Glaser Critical Thinking Appraisal. Accuracy in nursing assessment was measured by deviations between the patient's SDS observed score and the patient's SDS predicted score. The patient's predicted score was based on the relationship between the Depression Status Inventory and Self-Rating Depression Scale. Perceived patient self-disclosure was measured by a 3-point system on patients' disclosure of information on symptoms of depression. A Pearson product-moment correlation and multiple regression analysis were used to analyze the data. The result showed that nurses' education was not significantly related to accuracy in assessment but related to critical thinking ability (r = .26; p = .004). Nurses' critical thinking ability was significantly related to accuracy in nursing assessment depression (r = -.24; p = .008). Perceived patients self-disclosure was not significantly related to accuracy in nursing assessment of depression. PMID- 8707533 TI - Geriatric depression: a survey of nurses' knowledge and assessment practices. AB - A sample of 149 registered nurses were surveyed to determine their knowledge of the symptoms of depression in the elderly and their assessment practices. Respondents were able to identify not only the traditionally associated symptoms but also two other behaviors, irritability and blaming others, which are not usually designated by other health care providers as associated with geriatric depression. These additional observations may reflect nurses' providing extensive physical as well as verbal care over much longer time periods for patients in inpatient and home settings than do other professionals. Recent findings in nursing research support the presence of these other behaviors as symptoms of depression. An additional finding revealed that nurses were not assessing their patients for depression even though they had a knowledge base of the symptoms. PMID- 8707534 TI - Nursing postvention for spousal survivors of suicide. AB - There is a scarcity of information available with respect to postbereavement outcomes for survivors of the suicide of a loved one. Few studies have focused on postvention therapies for the bereaved, particularly the bereaved survivors of suicide. The major aim of this study as to compare the effects of two theoretically derived nursing postventions, Bereavement Group Postvention (BGP) and Social Group Postvention (SGP), among the widowed whose spouses died of suicide. The findings suggest that both groups experienced an overall reduction in depression and distress. Although participants in the SGP generally showed significant improvement in social adjustment, they tended to be less well adjusted with respect to their parental roles at the end of the 8-week postvention sessions. Comparison of the psychoemotional correlates of grief varied such that there were no significant differences between the postvention groups for social isolation, loss of control, somatization, or death anxiety. The BGP participants experienced significantly reduced levels of anger/hostility and guilt; however, feelings of anger/hostility actually increased for those receiving the SGP. There was a significant reduction in feelings of despair, rumination, and depersonalization for both groups. Although social isolation was not significantly reduced for participants in either group, those receiving the BGP tended to experience a reduction in social isolation and those receiving the SGP showed no changes. PMID- 8707535 TI - Menopause and hormone replacement therapy from holistic and medical perspectives. AB - Myths and other issues surrounding menopause are examined on the basis of historical and current literature from medicine, psychiatry, and psychiatric nursing, and on current research. Changes in the psychiatric view of menopause and mental illness reflect a more holistic view of menopause. Some effects of menopause during this normal transitional phase of a woman's life are explored with respect to the developmental, physiological, and cognitive/psychosocial domains. Concepts of menopause as disease or as normal development are discussed as well as issues related to "care or cure" interventions for menopausal women. Evidence supports the need for systematic longitudinal research studies on the use of hormone therapies to provide information on their long-term effects on the menopausal woman. The use of hormone therapies alone or in conjunction with other holistic interventions is discussed. Nurses have a professional responsibility to come to terms with the continuing conflict related to hormone therapies so that they may provide appropriate nursing interventions to celebrate this passage rather than deny it. PMID- 8707536 TI - Essential structure and meaning of recovery from clinical depression for middle adult women: a phenomenological study. AB - The author interviewed 22 women who had been clinically depressed and were in the recovery stage, as evidenced by their self-evaluations and scores on the Beck Depression Inventory. The focus of the interviews was on the phenomenological question concerning the meaning of the women's experiences of recovering from depression. Five phases in the recovery process were identified from analysis of the data: existential alienation/pain, crises of adulthood, first turning point, second turning point, and becoming a gardener. Nurses can use perceptions of clients in this study about what was helpful and not helpful and the tools they used to help themselves in recovering from depression to enhance the recovery process for other depressed women. PMID- 8707537 TI - Knowledge of depression and application to practice: a program evaluation. AB - The purposes of this program were to determine whether participants gained more comprehensive knowledge about depression and if that knowledge was applied in practice following attendance at a 3-day educational workshop. Twelve programs were conducted in three southwestern states. The majority of the 363 participants were Anglo-White female registered nurses whose ages ranged from 23 to 74 years and who worked in general medical-surgical units, nursing homes, or community health settings. This study was a repeated-measures design to evaluate the effects of depression education on depression knowledge and its clinical application. Participants demonstrated increased depression knowledge and practice application following the program. Knowledge of depression remained stable across postprogram measures. Although application of interventions decreased between 6 weeks and 6 months, assessment skills remained stable. Mental health nurses can improve patient outcomes through programs such as the one described by educating nurses in the detection and management of depression. PMID- 8707538 TI - As nurses, we embrace the notion of caring. PMID- 8707539 TI - Facilitative affiliation: nurse-client interactions that enhance healing. AB - Facilitative affiliation is a new concept developed to capture the essence of effective nurse-client interactions, regardless of the client population or the practice setting. This concept has been synthesized within the theoretical framework of Modeling and Role-Modeling. Defining attributes have been identified as (a) presence, (b) assessment of needs based on the client's self-care knowledge and perceived resources, (c) creative individualized interventions, (d) selective normative disregard, (e) mutual trust, (f) nurturance, and (g) advocacy: Facilitative affiliation is defined as any nurse-client interaction in which the nurse assesses the client's needs based on that individual's self-care knowledge and perceived resources and creates individualized interventions based on those identified needs. The relationship is then characterized by availability, nurturance, and advocacy on the part of the nurse and a sense of mutual trust between the client and the nurse. Several client outcomes have been posited including that of enhanced healing. PMID- 8707540 TI - Factors that influence the mother-infant dyad relationships and infant well being. AB - This article presents a theoretical model derived by synthesizing research findings relevant to maternal perceived support, maternal attachment, infant well being, and infant attachment. The model, depicting possible predictors that facilitate positive mother-infant relations as well as outcomes of the dyadic relationship, may be used to plan interventions to promote healthy mother-infant relationships. Implications for nursing practice are described. PMID- 8707541 TI - Transcending breast cancer: reconstructing one's self. AB - This article presents the author's personal journey through and beyond breast cancer. The holistic process of transcendence that results in an ongoing sense of reconstructing and integrating one's Self (mind, body, and spirit) is described. Numerous examples are provided of how listening to and trusting one's inner voice is integral to the process of transcendence and reconstruction. Nursing implications are identified along with theoretical perspectives for understanding the author's experiences. PMID- 8707542 TI - Uncertainty, spiritual well-being, and psychosocial adjustment to chronic illness. AB - This research was undertaken to assess spiritual well-being as an internal coping resource to buffer the effects of uncertainty on psychosocial adjustment among 94 persons with diabetes mellitus. Five instruments were used to collect data; descriptive, correlational, multivariate, and content analysis procedures were used for data analysis. A predicted negative relationship between uncertainty and spiritual well-being was supported, with a stronger relationship between the existential well-being component of spiritual well-being and uncertainty than the religious well-being component. Uncertainty explained 43% of the variance in psychosocial well-being; existential well-being added an additional 10%, suggesting that it serves as a buffer between uncertainty and psychosocial well being. The findings suggest that spiritual well-being may be an important internal resource for persons forced to adjust to uncertainty related to long term health problems such as diabetes mellitus. PMID- 8707543 TI - Medical devices and attachment: holistic healing in the age of invasive technology. AB - The environment is filled with wonderful examples of medical technology that provide emergency life support, improve well-being, and offer the possibility of a longer and more productive life. But these devices are no longer only a part of the external environment. As technology advances, more devices are becoming a part of the internal environment (i.e., our bodies) as well. For some people an implanted mechanical device, such as a cardiac pacemaker or a cardioverter defibrillator, is readily accepted; for others it may be seen as an encroachment. It may be a symbol of loss and debilitation or of independence and resilience. What makes the difference? How can nurses facilitate a healthy adjustment and healing in an era permeated with technology? A discussion of the symbolism, related theory, and nursing implications is provided. PMID- 8707544 TI - Affiliated-individuation in caregivers of adults with dementia. AB - The purpose of this study was to test the effect of a theory-based support group intervention on affiliated-individuation in caregivers of adults with dementia. Modeling and Role-Modeling, a theory and paradigm for nursing, provided the framework for intervention. The sample of 26 caregivers attended biweekly support groups for 12 months. Quantitative data, collected at baseline and at 6 and 12 months, were analyzed using MANOVA to assess within-subject variance. Results of data analysis indicated that levels of affiliated individuation did not decrease in the subjects in spite of their continued caregiving of persons who had Alzheimer's disease. Qualitative data collected 1-2 months after 12 months of interventions supported the quantitative findings. They also described a process that parallels the theoretical definition of affiliated-individuation. PMID- 8707545 TI - Conceptualization of perceived enactment of autonomy in the elderly. AB - The eudaemonistic model of health is congruent with nursing's primary aim of practice and nursing's ethical imperative to support client autonomy. Although autonomy has been linked to holistic health, the definition of autonomy has been imprecise. This is especially problematic when working with the elderly. For clarification, a concept analysis of autonomy was completed. This led to further refinement and development of a new concept, perceived enactment of autonomy. This definition represents the subjective perceptions of the ability to act autonomously. Interventions derived from the nursing theory, Modeling and Role Modeling (Erickson et al., 1983/1988), and supportive of perceived enactment of autonomy and environments conducive to healing within the eudaemonistic health paradigm are proposed. PMID- 8707546 TI - Self-transcendence: a resource for healing at the end of life. AB - An emerging body of theoretical and empirical knowledge links the concepts of self-transcendence and healing in persons facing end-of-life issues related to aging or life-threatening illness. Self-transcendence is postulated to be a resource for healing in that reaching beyond self-boundaries may lead to a sense of well-being derived from an intensified awareness of wholeness and integration among all dimensions of one's being. This paper proposes a link between self transcendence and healing, based on clinical and empirical literature. Suggestions for approaches nurses may use to facilitate self-transcendence are included. PMID- 8707548 TI - Optometrists threaten public safety with bill. PMID- 8707549 TI - How to become the positively perfect physician. PMID- 8707547 TI - PPD skin reactivity and anergy in HIV-infected patients in Hawaii. AB - This study was a prospective screening study for PPD and anergy skin test reactivity in 304 HIV-positive individuals. A PPD positivity rate of 4.1% and an anergy rate of 50.5% were observed. The Hawaii HIV population has a relatively low prevalence of latent TB compared with the high prevalence of TB in the Hawaii population at large. PMID- 8707550 TI - The role of cranial computed tomography in the diagnostic work-up of headache. AB - Approximately 18% of all patients referred for a cranial computed tomography complain of headache only. We reviewed 363 consecutive patients in order to assess the value of this examination in the diagnostic approach. Despite the vast number of normal examinations (88.4%), we advocate the routine use of a cranial computed tomography in every patient with chronic headache. The cost of the examination can significantly be reduced by performing an unenhanced scan only. An additional contrast-enhanced scan should be obtained if a suspicious lesion is seen. Brain magnetic resonance imaging is not indicated except in the preoperative workup of a lesion visualized on computed tomography. PMID- 8707551 TI - Chronic daily headache in children and adolescents: a 2-year prospective study. AB - Chronic daily headache in children and adolescents has not been well described. We analyzed data for 37 children and adolescents who presented with chronic daily headache to our Pediatric Headache Clinic over a 2-year period. These youngsters had five distinct headache patterns; 40% of them had the "comorbid" pattern, 35% were classified with new daily persistent headache, 15% with transformed migraine, 5% with chronic tension-type headache, and 5% could not be classified. There were no significant differences by diagnosis in externalizing and internalizing behaviors, type A behaviors, disability, pain severity, days missed from school, and number of coping skills employed. Children and adolescents with chronic daily headache have distinct clinical patterns, but for the most part, have similar disability. Differences between adult and childhood chronic daily headache are emphasized. PMID- 8707552 TI - The placebo effect in acute headache management: ketorolac, meperidine, and saline in the emergency department. AB - In a prospective, double-blind, randomized study, ketorolac 60 mg, meperidine 50 mg plus promethazine 25 mg, and normal saline given by intramuscular injection were compared as treatment for acute headache crises. Thirty patients (6 men and 24 women) presenting to an urban emergency department with any type of benign headache were randomized into three groups and filled out the McGill Short-Form Pain Questionnaire with a Pain Rating Index and a Visual Analogue Pain scale. They received one of the study medications and repeated the testing after 1 hour. The objective was to test the efficacy of ketorolac in this population. Separate analyses of the McGill Short-Form (Total, Sensory, Affective, and Pain Rating Index scales) and the Visual Analogue Pain scale responses showed that the three treatments produced a significant reduction in pain (P < .0001), but that pain reduction did not differ among the treatments. This profound reduction observed after administration of a placebo prevented accurate evaluation of the effects of ketorolac. The placebo response must be considered in the design of future trials using intramuscular medications in the acute intervention of headache crises. In addition, the use of a standard analgesic is necessary to demonstrate both assay sensitivity and magnitude of response to placebo. PMID- 8707553 TI - Electromyographical ischemic test and intracellular and extracellular magnesium concentration in migraine and tension-type headache patients. AB - Headache has often been described in the hyperexcitability syndrome which recognizes an alteration of calcium and magnesium status in its etiopathogenesis. Moreover, in migraine patients magnesium has been shown to play an important role as a regulator of neuronal excitability and, therefore hypothetically, of headache. The present research involves a neurophysiological evaluation and magnesium status assessment of a group of headache patients. Nineteen patients (15 women and 4 men) with episodic tension-type headache and 30 patients (27 women and 3 men) with migraine without aura were examined. An ischemic test was carried out on the right arm with electromyographic (EMG) recording of motor unit potential activity during the interictal period. The determination of extracellular (serum and saliva) and intracellular (red and mononuclear blood cells) magnesium was also performed. The EMG test was positive in 25 of 30 migraine patients and in 2 of 19 tension-type headache patients. Between the two patient groups, there were no significant variations in the concentration of extracellular and white blood cell magnesium, while the red blood cell concentration of this mineral in the group of migraineurs was significantly reduced with respect to that in the group of tension-type headache patients (P < 0.05). The positive EMG test was significantly associated with a low concentration of red blood cell magnesium (P < 0.0001). These results confirm previous findings by demonstrating different etiopathogenic mechanisms as the basis of migraine and tension-type headache. Migraine seems to be related to an altered magnesium status, which manifests itself by a neuromuscular hyperexcitability and a reduced concentration in red blood cells. PMID- 8707554 TI - The IHS classification criteria for migraine headaches in adolescents need minor modifications. AB - The operational and diagnostic criteria for migraine and all other headache disorders released in 1988 by the International Headache Society are universally considered reliable and exhaustive. These criteria, however, cannot be considered as satisfactory for population-based studies on migraine prevalence, especially if adolescents are the subjects of the study. Using these diagnostic criteria, we conducted an epidemiological study in order to assess the prevalence of migraine headache in a student population aged 11 to 14 years. Our survey made it possible to code IHS 1.1 (migraine without aura) in 2.35%, IHS 1.2 (migraine with aura) in 0.62%, IHS 1.7 (migrainous disorders not fulfilling migraine criteria) in 1.52%, and IHS 13 (headache not classifiable) in 1.38% of the examined pupils. In adolescents, the low prevalence estimates of migraine headache coded IHS 1.1 and the relatively high prevalence estimates of headaches coded IHS 1.7 and IHS 13 have appeared to be a consequence of the rigidity of some operational diagnostic criteria of the recent IHS classification rather than of the geographical, environmental, or socioeconomical peculiarities of the cohort. Therefore, in order to improve the reliability and the exhaustiveness of the IHS classification by increasing its sensitivity, we believe that minor modifications of the diagnostic criteria are necessary. Within these revised criteria, the subitem "moderate or severe intensity" of pain headache should become mandatory, whereas the lower limit of the criterion "duration of pain" should be reduced to 1 hour. PMID- 8707555 TI - The confirmation of a biochemical marker for women's hormonal migraine: the depo estradiol challenge test. AB - PRECIS: Will estrogen withdrawal cause migraines in post-menopausal women? OBJECTIVE: To record the changes in serum estradiol and total estrogen levels after an intramuscular estradiol injection in menopausal subjects and then record any subsequent migraine occurrence. DESIGN: Open selection process, comparative trial. PATIENTS: Twenty-eight postmenopausal women volunteers were given 5 mg depo-estradiol cyprionate as an intramuscular injection. Sixteen (migraine group) had a history of severe, cyclic, menstrually related migraine attacks before becoming menopausal. Twelve (control group) had no history of migraine or headache. All volunteers were on continuous estrogen replacement therapy at the beginning of the study. Progestins were not used in the study. MAIN OUTCOME MEASURES: Serum estradiol and total estrogen levels were measured prior to the depo-estradiol injection and on subsequent days 4, 7, 14, 21, and 28. RESULTS: Total estrogen and estradiol levels varied greatly at every measured interval. Menopausal complaints of vasomotor symptoms were relieved for at least the first 2 weeks of the study. No member of the control group reported a migraine during the month. However, a severe migraine was reported by all 16 women with a history of migraine. The average day of the migraine occurrence was 18.5 +/- 2.8. The serum level of estradiol on the day of the worst migraine was 46.4 +/- 5.6 pg/mL. The significance of these findings was at the 95% confidence level. CONCLUSIONS: This study confirms two factors about menopausal hormonal migraine: (1) it can be precipitated by a drop in serum estrogen levels, and (2) a period of estrogen priming is a necessary prerequisite. This study also identifies that there are two biologically different populations of postmenopausal women: (1) those who developed migraine after a single depo-estradiol injection, and (2) those who did not. By understanding that in addition to the biological predisposition to migraine there exists the biochemical cofactor of falling estrogen levels, we may better understand this phenomenon and develop means to prevent its occurrence. PMID- 8707556 TI - Cervicogenic headache: the influence of mental load on pain level and EMG of shoulder-neck and facial muscles. AB - The relationship between pain and EMG levels was studied in 17 cervicogenic headache patients and 17 group-matched healthy controls. All subjects performed a 1-hour, complex, two-choice, reaction time test. Every 10 minutes before, during, and also for 20 minutes after the test, they reported pain levels (using visual analogue scales) in the forehead, both temples, neck, and shoulders. Electromyographic activity, using superficial electrodes, was also recorded from the frontal, temporal, neck (splenius), and trapezius muscles. Maximal voluntary contractions were performed in all the muscles. Increased pain levels before, during, and after the test were found on the symptomatic side in the temple, shoulder area, and neck in the patient group compared with nonsymptomatic side and controls (neck only compared with controls). Electromyographic amplitudes from the trapezius muscle on the symptomatic side were significantly higher before and during the test, compared with the nonsymptomatic side, but most markedly during the test. Pretest EMG amplitudes from the frontal muscle on the symptomatic side in patients were also significantly higher than those in controls, but the difference vanished during the test. There are indications that the temporal pain, ie, the headache, is a referred pain. These observations may point to a "muscular" involvement in the pathogenesis of cervicogenic headache, either primarily or, which seems more plausible, secondarily. PMID- 8707557 TI - A community-based prevalence study on headache in Malaysia. AB - This community study on headache in Malaysia was based on IHS diagnostic criteria and showed the last-year prevalence of migraine was 9.0%. Migraine with aura accounted for only 10.6% of the migrainous population. The last-year prevalence of tension headache was 26.5% (94.4% episodic, 5.6% chronic) and 28.2% for other types of headache. No case of cluster headache was found. Almost two thirds of the migraine subjects graded their headaches as severe, while almost 60% of the tension headache subjects and almost 70% of the other headache subjects graded their headaches as mild. Overall, there was higher prevalence in females for migraine and tension headache, and in males for the other types of headache. The prevalence of headache was lower among those younger than 15 and older than 65 years of age. No significant differences were found in the prevalence of headache among the different racial groups nor among the urban versus the rural population. All the headache types shared the same triggering factors suggesting that different physiological characteristics are responsible for the type of pain suffered. In the location of this community with its tropical climate, headache was attributed to sun exposure in 51.9% of the migraine subjects, 55.7% of the tension headache subjects, and 36.6% of the group with other headaches. PMID- 8707558 TI - Prevalence of primary headache syndrome in adults in the Qassim region of Saudi Arabia. AB - BACKGROUND AND PURPOSE: Headache occurs worldwide, but documentation on the burden of headache in Saudi communities is quite limited. The aim of this study was to determine the prevalence of primary headache in a rural community in the Qassim region of Saudi Arabia. METHODS: A door-to-door survey of subjects older than 15 years of age was used for completion of a structured questionnaire to determine headache types based on defined criteria. Physical examination was used to exclude secondary causes. RESULTS: Of 5891 inhabitants, 473 suffered from headaches. The crude lifetime prevalence rate was 8% (95% Cl, 7.3% to 8.7%) and with age adjustment was 9.3% (8.6% to 10%). Headache sufferers were predominantly women. The age-specific rate increased progressively, peaking in the sixth decade. Tension-type headache diagnosed in 185 subjects predominated (3.1%), while the prevalence of migraine was 2.6%. CONCLUSION: The findings agree with the reported prevalence of 10% to 12% in other communities in the Kingdom of Saudi Arabia, but lower than the rates in Western countries. The lower rates are probably related to sociocultural factors. PMID- 8707559 TI - Use of high sumatriptan dosages during episodic cluster headache: three clinical cases. AB - The authors describe three patients with episodic cluster headache whose attacks were all treated with subcutaneous sumatriptan. The patients described had a high frequency of attacks (more than two per day); therefore, far higher dosage of the drug was taken than commonly used in cluster headache. The patients did not experience any particular side effects, neither during the treatment period nor on abrupt withdrawal of the drug. Moreover, neither tachyphylaxis nor addiction were observed. The authors point out both the efficacy of sumatriptan, confirmed in all the treated attacks, and its safety even at higher dosages than recommended. PMID- 8707560 TI - Cardiac arrest caused by trigeminal neuralgia. AB - A 67-year-old man with a 12-year history of trigeminal neuralgia experienced multiple fainting episodes preceded by right facial pain. One episode resulted in cardiac arrest with successful resuscitation. Pacemaker insertion prevented further episodes of syncope despite the occurrence of pain. The fainting episodes and cardiac arrest are believed to be unusual manifestations of trigeminal neuralgia. PMID- 8707561 TI - Acupuncture and migraine. PMID- 8707562 TI - A controlled trial of general practitioners' attitudes to patients with schizophrenia. AB - OBJECTIVE: To examine general practitioners' attitudes to patients with schizophrenia. DESIGN: A random sample of primary care physicians were alternately sent a case vignette of a patient with or without schizophrenia, in an otherwise identical clinical abstract, and asked to indicate their level of agreement with fifteen statements based on it. SUBJECTS AND SETTING: A one-in five sample of general practitioners who were identified from the Primary Care Services Register of Lothian Health Board. RESULTS: The median score for each statement was compared by the two-tailed Wilcoxon rank sum test. Doctors responding to the vignette of the patient with schizophrenia were significantly less willing to have the patient on their practice list, more likely to refer them to a specialist and more likely to think that they would be violent; whereas they did not think that they would take up any more time than the other patient. These impressions were no different between those who had or had not received work training in psychiatry. CONCLUSIONS: This controlled trial of primary care physicians' attitudes towards patients with schizophrenia amounts to an empirical demonstration of medical discrimination against the sufferers of this and potentially of other long term psychiatric disorders. Psychiatrists and general practitioners should share care in the management of schizophrenia and try to overcome the prejudices against such patients in an attempt to improve their overall clinical care. PMID- 8707563 TI - Using economics alongside medical audit. A case study of the management of endometriosis. AB - During 1993, the diagnosis and initial management of endometriosis in women was identified as an important topic to be addressed within the Gynaecology Audit Project in Scotland (GAPS). This paper documents the experience of using the medical audit data collected by this exercise to estimate the costs and outcomes associated with alternative treatments for endometriosis associated infertility. Its aim is not to draw firm policy conclusions but rather to demonstrate the economic methods and to highlight difficulties in the application of economics to audit. The average cost of medical management was found to be significantly higher than the average cost of expectant management because of the high costs of the drugs involved. There was little difference in clinical and health outcomes observed between those women managed expectantly and those women managed medically. The results of the economic assessment add cautious support to the growing volume of clinical evidence indicating expectant management as the first line treatment of choice although further research is required to substantiate these results in a larger sample of women presenting with this condition. PMID- 8707565 TI - The Measles/Rubella Immunisation Campaign in Greater Glasgow: a success in collaboration. AB - The recent UK campaign to immunise schoolchildren against measles and rubella involved, in Greater Glasgow, about 143,000 schoolchildren attending 300 primary, 63 secondary and 54 special schools administered by Strathclyde Regional Council and 21 private schools. The campaign involved a large number of individuals and professional groups, both in education and the health service. Despite-or perhaps because of-the magnitude and complexity of the task, and the very short time available, the campaign was planned and brought to a successful conclusion with a quite remarkable degree of enthusiasm and goodwill. As a result it was possible to immunise 88% of eligible schoolchildren in an area which has 46% of its population living in the most deprived postcode sectors (Carstairs categories 6 and 7) and to record all results on computer file as a permanent and readily accessible record. The campaign provided an unusual opportunity to demonstrate how effectively different organisations within the health service can collaborate with one another and with other agencies when there is a clear goal and deadline imposed. PMID- 8707564 TI - Referral pattern and management of patients with malignant brain tumours in south east Scotland. AB - We reviewed hospital case notes of all incident cases of intracerebral tumours in SE Scotland to analyse referral pattern and time intervals between presentation, diagnosis and treatment. We identified 439 new patients with intracerebral tumours in a two year period: 64% single (primary brain tumours 43%: metastasis 21%) and 36% multiple (metastases). Ninety-two per cent of patients were referred by GP's and 88% were self referrals to hospital or were initially identified at hospital follow-up clinics. Only 27% of patients were initially referred to the specialist centre (Western General Hospital) but 57% were CT scanned at the specialist centre and 83% were referred to there at some time. Time from GP referral to CT scan was related to availability of local CT scanning. Only 10% of cases with probable single metastases and 39% with HGG were treated with surgery plus radiation. Cranial irradiation was started within four weeks of CT diagnosis in 79% of those with metastases but in less than 5% of patients with HGG. The patterns of care for patients with brain tumours show great variation. Hospital referral guidelines, better inter-hospital and inter-department communications and more available access to CT scanning and radiotherapy should improve the quality of care and possibly treatment outcome in this group of patients. PMID- 8707566 TI - Strengths and limitations of using the Revised Elderly Person's Disability Scale (REPDS) as a research tool in a Scottish context. PMID- 8707567 TI - Multiple myeloma in north east Scotland: a review of incidence and survival over three decades. AB - STUDY OBJECTIVE: (1) To review the pattern of published world age-standardised registration rates for myeloma (ICD8 and ICD9 203) for the five Scottish regional cancer registries between 1973-77 and 1983-87. (2) To review the patterns of world age-standardised incidence and survival for myeloma in Grampian region over the time period 1960-89. DESIGN: Retrospective analysis of cancer registration data and linked mortality data. SETTING: The five Scottish regional cancer registries (East, North, North East, South East and West Scotland). PATIENTS: Incidence: a total of 405 patients with myeloma resident in Grampian region (153 diagnosed between 1960-69, 252 diagnosed between 1980-89 inclusive). Survival: a total of 420 patients with myeloma treated in Grampian hospitals between 1968 and 1987 inclusive. MEASUREMENTS AND MAIN RESULTS: On average, the world age standardised registration rates for the five regional registries increased from 2.8 to 3.1 cases per 100,000 in males and from 1.8 to 2.4 per 100,000 in females between 1973-77 and 1983-87. No clearcut pattern in the trends for individual registries was evident for males but, in general, an increasing trend in female rates was observed for the registries with the lowest rates initially, whilst those with the highest initial rates increased only slightly or even fell. After age and sex standardisation, the annual incidence of myeloma in NE Scotland has increased by 20 per cent between 1960-69 and 1980-89, from 2.4 to 2.9 cases per 100,000 population per year with a disproportionate increase in older patients. Between the two time periods female rates remained stable or increased over all age groups while male rates fell for ages under 69 years and rose for ages above this, a pattern which was reflected in changes in the male:female ratio. The five year survival rate for all ages (14%) has not improved since 1968-72 similar to overall Scottish figures. In NE Scotland, younger patients appear to fare better, and older patients worse, compared with the overall Scottish experience. CONCLUSIONS: The increase in myeloma incidence may be due to a combination of improved ascertainment, especially in the elderly and a possible true increase in incidence in females, suggestive of increased exposure to an aetiological agent in the past. A formal year birth cohort analysis is required to confirm this finding. The overall prognosis for myeloma remains poor, especially for elderly patients and efforts to elucidate the aetiology must continue. PMID- 8707568 TI - Health check-ups in general practice: a patient perspective. AB - One of the strategies identified by the NHS to achieve improvements in the health status of the Scottish population is to implement health promotion programmes through the general medical practitioner service. Grampian Local Health Council undertook a survey of Grampian residents to identify factors which might encourage greater public participation in such programmes. The results which are analysed and discussed in this paper show that factors identified by the public include: a preference for doctors to carry out health checks; the need for public education on the roles of different health professionals; the need for information on health checks and their benefits to be supplied in advance; a choice of appointment times and the offer of a follow-up appointment to discuss test results. These results are of interest to health professionals as they are likely to contribute to higher levels of patient involvement in health promotion programmes. PMID- 8707569 TI - Palliative care experience and training of Scottish general practitioner trainees. AB - OBJECTIVES: (a) To establish the clinical experience of Scottish general practice trainees in palliative care. (b) To establish the extent of their training in the discipline. (c) To elicit their views on home care. DESIGN: Postal questionnaire survey. SUBJECTS AND SETTING: Forty-seven trainee general practitioners attending the Scottish National Trainees Conference in Aberdeen in February 1995. MAIN OUTCOME MEASURES: Experience of palliative care in hospital and in general practice and training received in each setting. Perception of need for further training. Views on home care. PMID- 8707570 TI - Medical audit activity in primary and secondary care in the west of Scotland. AB - OBJECTIVES: To look at the level of activity and motivation towards audit in primary and secondary care in the West of Scotland. DESIGN: An anonymised postal questionnaire survey. SETTING: Area covered by six Health Boards in West of Scotland. SUBJECTS: 150 GPs and 150 hospital-based clinicians randomly selected from two departmental databases. RESULTS: There appears to be a rule of diminishing returns in operation where half of those involved in audit have completed a project and half again have repeated a project. The current level of activity is significantly higher in secondary than primary care (chi 2 p = 0.05). There is significantly more audit activity in teaching than non-teaching hospitals (chi 2 p = 0.01) and significantly more GPs from larger practices have been involved in audit than those from smaller practices (chi 2 p = 0.05). All of those involved in audit have set standards. Levels of motivation towards audit were significantly higher amongst hospital-based clinicians than general practitioners (chi 2 p = 0.001). CONCLUSIONS: There has been a widespread involvement in audit in both primary and secondary care in the West of Scotland, but activity and enthusiasm or motivation are lower amongst general practitioners. PMID- 8707571 TI - Lothian general practitioners, HIV infection and drug misuse: epidemiology, experience and confidence 1988-1993. AB - BACKGROUND: Lothian has a high prevalence rate of HIV infection associated particularly with injecting drug use in the mid-1980s. During the late 1980s a number of services were introduced to support general practitioners to care for drug users and people with HIV infection. AIM: To ascertain the practice distribution of patients with HIV infection and drug users, and the experience and confidence of Lothian GPs in a number of HIV and drug related tasks, and to see how these changed over a five year period after the introduction of support services. METHOD: Information about practice distribution of patients with HIV infection and drug users and information about GP experience and confidence was obtained from a survey of individual GPs in 1988. Two practice surveys in 1991 and 1993, using a chosen GP for each practice, gave information about practice distribution of people with HIV infection and drug users, while a second survey of individual GPs in 1993, concentrating on GP experience and confidence, acted as a follow up to the 1988 survey. RESULTS: GPs are in touch with an increasing number of people with symptomatic HIV infection. They are increasingly involved and confident in caring for drug users. Offering support and training to GPs appears to have encouraged their involvement. There are a significant number of people with HIV infection in Lothian who are not known to GPs or to hospital services. A combination of practice and individual GP surveys was found to provide the most reliable information. It is planned to repeat the practice survey in 1995. PMID- 8707572 TI - Developing a local protocol for the management of lipid disorders in general practice. AB - Guidelines and protocols are increasingly becoming part of routine clinical practice. A standardised methodology has been proposed for the development of national guidelines based on principles which are known to effect validity. However, less is known about the most appropriate methodology for the development of local protocols. A case study is presented of the development of a local protocol for the management of lipid disorders in primary care in Grampian. It highlights the practicalities of such a process and identifies lessons for others who may wish to develop local protocols. PMID- 8707573 TI - The chief scientist reports.... Prevalence of psychiatric disorder, cognitive impairment and functional disability among homeless people resident in hostels. AB - This cross-sectional survey was performed to estimate the prevalence of psychiatric disorder among homeless residents of hostels in Edinburgh and to relate it to functional impairment. Full diagnostic assessments were completed on 136/198 (69%) of the subjects. Thirty-one per cent of subjects (95% CI 25, 42) had significant functional impairment and this was most strongly associated with the presence of functional psychosis and cognitive impairment. Few of the cognitively impaired subjects were in contact with health or social services. PMID- 8707574 TI - Death by suicide in Grampian 1974-1990', Health Bulletin 1996; 37-44. PMID- 8707575 TI - Trends in elderly patients (65 and over) waiting for long stay care in Edinburgh General Hospitals 1988-1994', Health Bulletin 1995; 373-8. PMID- 8707576 TI - [Increase in syphilis in the Alp, Donau, Moldau region]. PMID- 8707577 TI - [Raynaud phenomenon and scleroderma]. AB - The Raynaud phenomenon occurs in 90-100% of patients with systemic scleroderma. It is caused by dysregulation of the vascular tone between vasoconstrictive and vasodilatory influences. Early damage to the vascular endothelium and the activation of platelets lead to the release of vasoactive substances. Structural and function impairment of the perivascular neural fibers contributes to the vasospasm. New vasodilatory treatment strategies, e.g. infusions with calcitonin gene-related peptides, could be useful in the control of vasospasm in the peripheral extremities as well as internal organs in systemic scleroderma. PMID- 8707578 TI - [Human herpesviruses 6 and 7. Basic principles and possible significance for dermatology]. AB - Primary infection with human herpesviruses 6 and 7 (HHV-6 and HHV-7) during early childhood causes permanent latent infection, usually without any ill effects; only a small percentage of primary infections will lead to exanthem subitum. Like other herpesviruses. HHV-6 and HHV-7 can be reactivated at any time if host defence mechanisms become defective (e.g. in transplant recipients, AIDS, tumour patients). HHV-6 can be reactivated under such conditions and cause a variety of clinical problems, such as exanthems along with interstitial pneumonia or hepatitis for example. In addition, the reactivated virus may influence the course of autoimmune and proliferative diseases such as systemic lupus erythematosus and Hodgkin's disease. While, HHV-7 may be associated with similar disorder, more systematic studies are needed to clarify the clinical implications and the pathogeetic mechanisms of both viruses. PMID- 8707579 TI - [Women in dermatology. German and other European clinics in a comparison]. AB - The situation of women in dermatology was evaluated with the aid of questionnaires in spring 1994. A questionnaire was sent to 33 clinics in Germany and 46 in other European countries, asking about the proportions of male and female physicians in each department. In addition, individual department members were asked in another questionnaire about their reasons for choosing dermatology and about possible sex discrimination. The clinic questionnaires were returned from 22 German and 37 other European clinics and the individual questionnaires by 548 German physicians and 1164 physicians from the other countries (overall 52% women). We found that 41.8% of the dermatologists in German clinics were women, as against 43.4% in the other countries. Women made up about half of the fully qualified specialists, but accounted for only a small percentage of those in higher positions (10.8% in Germany, 27.9% in other countries). Of the 1712 physicians who answered the individual questionnaire, the majority had chosen dermatology because of their interest in the specialty. Slightly more women than men gave other reasons, such as more free time--mostly for family matters--or less physical strain. Clearly more women than men felt they were discriminated against professionally because of their sex. The reasons for this related primarily to the biological and social role of women in society. Overall, women are thus well represented in our specialty, except in leading positions. PMID- 8707581 TI - [High frequency color coded duplex ultrasound of small finger arteries--new diagnostic possibilities? A noninvasive method for visualizing digital circulation]. AB - Small vessels with luminal diameters of 0.2-2.0 mm have not been studied with routine ultrasound diagnostic procedures. Modern color Doppler imaging technology, however, should allow visualization of the blood flow even in such small vessels. The present study was therefore aimed at demonstrating the possibilities of examining small arteries in the fingertips of healthy volunteers with color Doppler imaging. The question was whether or not a standardized visualization was possible. We found that small arteries and their blood flow could be demonstrated in all (n = 10) healthy subjects. Furthermore, we demonstrated pathologic findings in the fingertip arteries of a patient with thrombangitis obliterans as an example of the diagnostic value of these new procedures. PMID- 8707580 TI - [Antipruritic effect of antihistaminic and local anesthetic topical agents after iontophoretic histamine stimulation]. AB - No adequate topical therapy is available for pruritus. As little is known about the local influence of antihistamines and topical anaesthetics on the pruritic effect of histamine, we studied these agents in 12 volunteers. The antipruritic effect of 15-min topical application of dimethindene maleate (Fenistil gel) and different agents (Optiderm, EMLA, Xylocaine-Salbe 5%) on subsequent focal histamine stimulus (20 mC) given by iontophoresis was evaluated. The results were compared with those of pretreatment with the corresponding placebo creams and observations on skin. Wheal and flare areas were evaluated planimetrically. Itch or pain ratings were entered on a scale every minute over a 24-min period. The examination also comprised alloknesis, i.e. elicitation of perifocal itch sensation by usually non-itch-inducing (e.g. mechanical) stimuli. Remarkably, all topically applied substances, regardless of antihistaminic or anaesthetic potential, reduced the area of alloknesis significantly. This is likely to be a result of diminished excitability of the cutaneous mechanoreceptors. Itching was significantly reduced by all active substances, including the placebo cream corresponding to Optiderm, which might be due to the presence of urea. PMID- 8707582 TI - [Dermatologic symptoms of parvovirus B19 infections]. AB - Infections with parvovirus B19 are often characterized by the well-known erythema infectiosum, usually accompanied by fever and arthralgia. Furthermore the virus may cause a lethal hydrops fetalis during the pregnancy or a severe aplastic crisis in patients suffer from chronic hemolytic anemia. We present three patients infected by parvovirus B19. Whose skin lesions included a non-specific exanthem, erythema multiforme, and purpura. The virus infection was confirmed both by serology and PCR identification. In conclusion the diagnosis of acute exanthems should include the search for virus infections. PMID- 8707583 TI - [Tufted hemangioma. Clinicopathologic and immunohistologic analysis of 5 cases of a distinct entity within the spectrum of capillary hemangioma]. AB - Two female and three male patients with acquired "tufted angioma" are presented. The age of these patients ranged from 10 to 62 years. Two lesions were sited in the head and neck region, two in the upper extremities, and one on the trunk. Clinically, the angiomatous lesions appeared as elevated plaques, flat lesions with papular and macular areas, or erythematous plaques with small nodules. In four cases a biopsy was done, and in one case the tumour was excised. Histologically, the neoplasms were characterized by irregularly distributed vascular tufts in the dermis, and, in one case, in the upper subcutis. The vascular tufts were composed of plump endothelial cells and spindle-shaped pericytes surrounded by crescent-shaped vascular spaces. The positive staining for CD 31 and for CD 34 and alpha-smooth muscle actin, and the negative staining of endothelial cells for factor VIII underline both the existence of two cellular components in tufted angioma and the immaturity of endothelial cells. Evidence of regular mitotic figures in two cases and increased proliferative activity in three out of four cases tested, emphasize the neoplastic nature of slowly growing tufted angioma. Benign tufted angioma is a distinct entity in the spectrum of capillary haemangiomas and must be distinguished from other vascular neoplasms. PMID- 8707584 TI - [Cyclosporin A therapy of chronic recurrent polychondritis]. AB - We report on a 71-year-old female patient with relapsing polychondritis in both ears. Monotherapy with low-dose ciclosporin led to a complete remission. PMID- 8707585 TI - [Eruptive vellus hair cysts]. AB - Eruptive vellus hair cysts are rare epidermal cysts with vellus hairs, which appear in children or young adults. A 21-year-old patient is reported who developed asymptomatic comedo-like papules on his face, neck and chest. Histopathologically, epidermoid cysts with a fine stratum granulosum, laminated keratinous material and numerous vellus hairs were found in the mid-dermis. Evaluation of immediate family members revealed that the patient's father was similarly affected. Eruptive vellus hair cysts and steatocystoma multiplex are clinically similar, but have distinctive histopathological features and should be regarded as distinct entities. PMID- 8707587 TI - [Congenital familial plaque-shaped glomus tumors. An unusual variant of multiple regional glomus tumors]. AB - Plaque-like glomus tumours are an unusual and very rare clinical form of multiple regional glomus tumours. We report on a 16-year-old girl with this variety of glomus tumours. The irregular angiomatous plaques localized on her back and right hip were present from birth and enlarged with body growth. In addition, there were some bluish nodules scattered on the right mamma, the abdomen and the left shoulder. The family history showed hereditary influences. The proper diagnosis was based on histological and immunohistochemical findings. In the present case report, the clinical, histopathological and immunohistochemical features and the differential diagnosis of multiple plaque-like glomus tumours are discussed. PMID- 8707589 TI - [Diagnostic procedures in suspected immediate type allergy to penicillins]. PMID- 8707588 TI - [Tar particle inclusions and carcinogenesis]. PMID- 8707586 TI - [Hypothenar hammer syndrome as a rare cause of Raynaud syndrome]. AB - A 25-year-old carpenter with severe localised Raynaud syndrome of the lateral three fingers of his right hand is described. The patient reported digital ischaemia caused by cold or vibration, and especially when both were present simultaneously. He was initially diagnosed with thrombangitis obliterans, but failed to respond to intravenous prostaglandin therapy, pentoxifyllin or aspirin. Diagnostic procedures performed in our hospital, especially angiography, confirmed our suspicion of hypothenar hammer syndrome. Radiological examination revealed severe anatomical and functional changes of the distal ulnar artery and, digital arteries of the affected digits. The differential diagnosis of Raynaud syndrome should include the hypothenar hammer syndrome as a possible cause. Early diagnosis and prevention of hand trauma is the only possible way to stop progression of this disease, which leads to invalidity. PMID- 8707590 TI - [Kaposi sarcoma in Caucasian women. Clinical, chemical laboratory and endocrinologic examination of 8 women with HIV associated or classical Kaposi sarcoma]. PMID- 8707591 TI - [2nd Darmstadt Live Symposium for Surgical Dermatology 16-18 November 1994]. PMID- 8707592 TI - [Metal-induced dermatoses]. PMID- 8707593 TI - [Communications by the German Society of Dermatology. Comments and opinions on important problems in dermatology. Help yourself, then politics will help you]. PMID- 8707594 TI - Managed healthcare and the home healthcare nurse in the 1990s: selected legal implications. PMID- 8707595 TI - The stay or stray phenomena: reflections of home care aide fulfillment. AB - There is a growing need for home care services, which requires an increased dependable work force. This paper describes the development of a Structural Interview Guide used to determine indicators of job satisfaction of home care aides. By exploring with home care aides the factors that lead to job satisfaction, home care agencies can design programs to enhance satisfaction and thus retain proficient workers. PMID- 8707596 TI - Problematic standards: staff rights and nutritional assessments. PMID- 8707597 TI - Panic disorder: the overlooked problem. AB - The anxiety syndrome of Panic Disorder shares symptoms of numerous physical conditions and may aggravate existing physical conditions. Because many clients experiencing this disorder become isolated and homebound, the home care nurse is in a position to see patients with this disorder. This article provides assessment criteria guidelines that help the nurse identify patients with this problem. PMID- 8707598 TI - Is there life after hospital nursing? PMID- 8707599 TI - Increasing self-confidence through self-talk. AB - There are varied opinions on the commonness of self-talk. Some feel self-talk is engaged in more by persons who feel insecure and question their performance (Cauchon, 1994). Others feel self-talk is a process people use continually to describe and interpret the world, accurate or inadequate as the case may be. (Hansen, Rhode, and Wolf-Wilets, 1991; Braiker, 1989). Regardless of how much self-talk the nurse is aware of in interacting with clients and intrapsychically, it is important to assess thoughts for their logic or illogicality. Illogical thoughts should be replaced by positive words of encouragement, which will in turn raise the client's or nurse's self-concept and level of self-confidence. Making self-talk positive is always good nursing practice. PMID- 8707600 TI - Total parenteral nutrition in the older patient. PMID- 8707602 TI - Change and our home care paradigms. PMID- 8707601 TI - Applying advanced directives regulations in home care agencies. AB - As Dombi (1991) indicated, home healthcare agencies see people in their own settings and can be most influential in assisting clients and families to deal with important issues. Furthermore, in receiving care in their homes, clients and their families may be in a more amenable frame of mind to consider such issues. Agencies may find it helpful to identify special resource people who could be accessible to clients and families in enabling them to prepare appropriate documents. In a tape recording that reviews the state of life-sustaining therapy, Wolf (1993) highlighted the importance of seeing the regulations on advance directives as just the beginning of further work and discussion with clients and their families. Home healthcare agencies, in particular, need to do more than just comply with directives related to the Patient Self-Determination Act. Agencies can be in the forefront of identifying needs and concerns of clients and families related to critical life and death decisions and in exploring the ramifications of those decisions. The trend toward developing ethics committees in home healthcare agencies can be a vehicle for further exploration of such issues. PMID- 8707603 TI - Caring for the patient with laryngeal cancer at home. AB - Clients experiencing various stages of laryngeal cancer frequently are cared for at home with the help of home healthcare nursing and other agency personnel. This article provides the clinician with an organized way of assessing and intervening with these clients and their families. Strategies, practical tips, and hints are given to assist the nurse in providing high-quality care. PMID- 8707604 TI - Building trust during the initial home visit. PMID- 8707605 TI - The importance of student home care experiences. PMID- 8707606 TI - A telephone reassurance program for elderly home care clients after discharge. AB - Telephone reassurance programs (TRP) was developed to address the ongoing needs of chronically ill clients after home care discharge. Graduate students from Yale University School of Nursing worked with the Regional Visiting Nurse Agency of Connecticut, Inc., to develop this interesting program that could be replicated by home health agencies. Project objectives, a TRP protocol, program tools, potential benefits, and implications for agencies are presented. PMID- 8707607 TI - Management and evaluation of a care plan. PMID- 8707608 TI - The challenges of assessing skin indicators in people of color. AB - Expert physical assessment skills are critical in the practice of home healthcare nursing. This article describes ways nurses can assess and document skin indicators in people of color that are meaningful to everyone involved in the patient's care. PMID- 8707609 TI - The home care client with Alzheimer's disease: Part I: Assessment tools for activities of daily living. PMID- 8707610 TI - Diabetes nutrition therapy in homecare. AB - A large proportion of a home care nurse's caseload is elderly persons with diabetes, presenting as either a primary or secondary diagnosis. This article provides specific guidelines for nutritional knowledge and assessment of these clients while giving practical guidelines for nutrition counseling and education. PMID- 8707611 TI - Medicare managed care. PMID- 8707612 TI - Home health, AIDS, and refusal to care. AB - Refusing to provide nursing care to patients with human immunodeficiency virus (HIV) and acquired immune deficiency syndrome (AIDS) is an issue with complex ethical, moral and legal implications. As the number of patients with a diagnosis of HIV and AIDS increases in rural and urban areas, nurses and other healthcare providers must prepare to care for this growing population requiring home care. PMID- 8707613 TI - Tips to improve communication. PMID- 8707614 TI - The challenge of fall prevention in home care: a review of the literature. AB - Two thirds of accidents of people older than 65 years are due to falls, and a large proportion of death due to injury in the elderly can be traced to falls. Caring for people in their home environment allows home care nurses to have a pivotal role in the assessment of risk factors for falls and in the implementation of interventions that address primary, secondary, and tertiary activities that can prevent falls from occurring. This article provides an extensive review of the literature regarding falls and outlines successful clinical nursing interventions that can be used to prevent falls. PMID- 8707615 TI - Predictors of caregiver health strain. AB - Home care nurses work with family caregivers daily. Measuring the strain involved in the role of adult children as caregivers for their elderly family members at home is the basis of this study. Results identify the decline in health reported by caregivers and give the home healthcare nurse guidance in developing interventions for the important population. PMID- 8707616 TI - A creative way to use clinical nurse specialists in home care. PMID- 8707617 TI - A comparative study of fatigue complaints among assembly line workers employed in the two electronic factories in Vietnam. AB - A cross-sectional study concerning working conditions and the fatigue complaints of assembly line workers employed in two different electronic factories (A and B) in Vietnam was conducted from August to September 1994. While general working conditions, such as noise, dust, heat and lighting, were worse in Factory B, the prevalence rate of the subjective fatigue after working time was significantly higher among workers in Factory A. Fatigue symptoms in category I (Drowsiness and dullness) were particularly apparent among workers in Factory A. More interestingly, one-third of workers in Factory A complained of stiff shoulders and low back pain. Our field observation results suggest that the ergonomic inappropriateness of the assembly line in Factory A, may be causing a high rate of subjective fatigue among workers. PMID- 8707618 TI - Acute health hazards of firefighters after fighting a department store fire. AB - The purpose of this study was to evaluate the health hazards of firefighters after fighting a department store fire which lasted for 40 hours. Respiratory symptoms of 168 firefighters were collected and the pulmonary functions of 149 firefighters were measured by spirometer two days after fighting the fire and compared to 32 controls. The principal symptoms manifested by firefighters were burning eyes and mucous membrane irritation. Cross-sectional study of the pulmonary functions showed that FEV1.0, FEV1.0/FVC and the flow rates (MMF, FEF25%, and FEF75%) in smoking exposed firefighters were significantly lower than those in smoking controls. However, only FEV1.0 (%) was significantly lower in nonsmoking exposed firefighters than in nonsmoking controls. Both FVC and FEV1.0 showed a declining trend with the duration of fire fighting. This study provides further evidence that firefighters are exposed to irritants during fighting a fire. Exposure to the combustion products could lead to pulmonary function defects, especially in smoking firefighters. PMID- 8707619 TI - Direct quantitative analysis of respirable cristobalite on filter by infrared spectrophotometry. AB - A direct on-filter method in infrared spectrophotometry was investigated for the quantitative analysis of respirable cristobalite. A polypropylene membrane filter was employed for this analysis, because the membrane filter has high transparency and no interference peak in the spectral range to be used (750-250 cm-1). Linear relation between the sample weight and peak height (absorbance) for three specific peaks at 620, 385 and 300 cm-1 of cristobalite were confirmed over the range from 45 to 1,000 micrograms/cm2 for 620 cm-1 peak and 45-2,380 micrograms/cm2 for 385 and 300 cm-1 peaks. The variation of absorbance with the difference of particle size was smaller in filter sample than in potassium bromide pellet sample. As a conclusion, this direct on-filter method can be used for quantitative analysis of cristobalite in airborne dust in working environment. PMID- 8707620 TI - Pollution and its control in asbestos milling processes in India. AB - An intervention study has been conducted in chrysotile asbestos milling processes with an interval of one year. The 'Membrane Filter Method' adopted in the former study for fiber concentration estimation has been followed in the present study, to estimate the environmental samples under the same experimental conditions. Though very high fiber levels compared to the prescribed permissible limit in India, have also been obtained in this study, a general reduction trend in the values of mills common to the both studies are observed. A further suitable control measure has been recommended in the paper to reduce fiber levels in the milling processes. PMID- 8707621 TI - Calcutta metro: is it safe from noise pollution hazards? AB - A modest assessment of noise was made in Calcutta Metro, India's first ever underground tube rail system, to examine if the range of noise levels present could endanger the hearing sensitivity of workers for the Metro. Sound measuring instruments of a sound level meter, an octave band analyzer, and a sound level calibrator were used for measuring the sound pressure levels in platforms of three stations: Esplanade, Kalighat and Tollygunge. The results indicated that the averaged A-weighted SPLs in these stations were in the range of 84-87 dBA. In the coaches of the moving train the Leq values ranged 92-99 dBA and LNP 105-117 dBA, all exceeding the safe limit of day time noise exposure of 55 dBA and 85 dBA of ACGIH. The SPLs at 4,000 Hz in the coaches were also in excess of safe exposure limit of 79 dB. The findings thus posed a potential threat to the workers. PMID- 8707622 TI - Cold-induced vasodilatation response of finger skin blood vessels in older men observed by using a modified local cold tolerance test. AB - The time of onset and magnitude of cold-induced vasodilatation (CIVD) in six older men (62-70 years) were compared with those of seven younger men (20-29 years) by using a modified local cold tolerance test. This test consisted of a 10 min immersion of the left middle finger in cold water at 10 degrees C, and was substituted for a conventional test (30-min immersion in ice water at 0 degrees C). Vasodilatation occurred significantly later in the older group, and the magnitude of their response was significantly lower during the immersion. No vasodilatation occurred in two of the older men. As the finger skin temperature before the test was not significantly different between the two age groups, the difference of CIVD response in the two age groups was thought to reflect the age related changes of peripheral vascular reactivity to a local cold stimulus. Considering the fact that no subjects complained a great deal of cold pain during the immersion, our modified local cold tolerance test seems to be a useful method for detecting the age-related degradation of local cold tolerance and peripheral vascular reactivity in older workers. PMID- 8707623 TI - A reference value for delta-aminolevulinic acid in plasma in the population occupationally unexposed to lead. AB - To obtain a reference value for delta-aminolevulinic acid in plasma (ALA-P), we determined ALA-P levels in 141 subjects (105 males and 36 females) without occupational exposure to lead. The distribution of ALA-P levels in males can be regarded to follow log-normal distribution and the geometric mean was 8.7 micrograms/l with 95% confidence interval of 6.0-12.5 micrograms/l. These data would be useful as a baseline for evaluating the relationship between ALA and its biological effects. PMID- 8707624 TI - [Breathing mode and growth of the facial bones]. PMID- 8707625 TI - [Paroxysmal positional vertigo]. PMID- 8707626 TI - [Effect of breathing mode and nose ventilation on growth of the facial bones]. AB - Correlations between breathing mode and craniofacial morphology were investigated in 47 children at the ages of 6-15 years (average, 9.9 years). Apart from history and clinical examination, nasal endoscopy, rhinomanometry and measurements of cephalometric radiographs were included in the analysis. After separating the patients into a "normal face" group and a "long face" group by measuring the angle between the frontal skull base and the mandibular plane, we analyzed the data in attempting to correlate nasal obstruction with craniofacial development. Findings demonstrated a significant predominance of mouth-breathing compared to nasal breathing in the vertical growth patterns studied. Furthermore, significant differences were found during nasal endoscopy in the growth pattern and were attributed to large adenoids. These findings were confirmed separately in all patients up to the age of 9 years, but differences were clearer. Rhinomanometry and planimetric measurements of the sizes of the adenoids in craniofacial radiographs showed no unambiguous differences between the patient groups. Our present study was not able to establish clear causal correlations between mouth breathing, craniofacial development and adenoid size, suggesting that existing genetically determined craniofacial growth patterns are modulated by exogenic influences. Nonetheless, our results show a correlation between obstructed nasal breathing, large adenoids and vertical growth patterns. PMID- 8707627 TI - [Tragus perichondrium-cartilage island transplant in middle ear surgery. Method and results after 5 years]. AB - Temporalis fascia remains the most widespread material for reconstructing the tympanic membrane in tympanomastoid surgery. If total or partial ossicular replacement prostheses are needed or pathology of the eustachian tube causes ventilation impairment, a more rigid transplant material is required. Between 1989 and 1994 perichondrium-cartilage composite grafts were used in a series of 597 cases of tympanomastoid surgery performed at the University ENT Clinic of Wurzburg. The graft was taken from the tragus and prepared as a cartilage island with perichondrium attached to one side, the perichondrium-cartilage island transplant (PCI). If reconstruction of the ossicular chain was necessary, glass ionomer cement protheses (IONOS) were used. Closure of the tympanic membrane could be achieved in 90% of all cases. In those cases where ossicular chain reconstruction was postponed to a second procedure, closure of the drum was achieved in 96%. The audiological results of the different type III procedures were evaluated. By using the PCI technique a favorable closure of the air-bone gap to 10-25 dB was achieved, even in cases with advanced ear pathology technique and results are presented in detail. PMID- 8707628 TI - [Suprahyoid pharyngotomy for surgical therapy of malignant and benign oral and hypopharyngeal tumors]. AB - Many cases of oropharyngeal and hypopharyngeal neoplasms without diffuse infiltration of the larynx or mandible cannot be treated effectively by a transoral approach. In such cases a lateral and/or median translingual pharyngotomy can permit effective surgical therapy. However, these surgical techniques require greater effort and violate uninvolved tissues, such as the lip, mandible and floor of the mouth. In contrast to this, a suprahyoid pharyngotomy can be a simple and precise approach to the oropharynx and hypopharynx, and provide the shortest distance to the pathological process. The excellent exposure given to the oropharynx and hypopharynx offers a more exacting macroscopic identification of tumor margins and minimizes possible injuries to vital neurovascular structures. The wound created can usually be closed primarily without the need for regional flaps. These factors allow a faster healing of the wound and better rehabilitation, as well as avoiding delays in postoperative radiation therapy. Over the past 6 years a suprahyoid pharyngotomy was performed in eight patients. Five patients underwent resections of an oropharyngeal cancer while three patients required resections of benign neoplasms of the tongue base. In the cases of oropharyngeal cancer, a suprahyoid pharyngotomy was performed in combination with a unilateral or bilateral neck dissection. A tracheotomy was required in six patients. Three patients underwent postoperative radiation therapy. No locoregional recurrences were found in these patients, with a median follow-up of 20.8 months. PMID- 8707629 TI - [Pathologic mechanoelectric transduction of outer hair cells as the cause of recruitment]. AB - It is believed that the sound-induced travelling wave in the mammalian cochlea is enhanced and sharpened by a positive feedback mechanism. This causes the passive linear basilar membrane growth function to become non-linear. The present paper shows that nonlinear basilar membrane vibration is due to the nonlinear growth function of the receptor potential of outer hair cells, which can be described by a 2nd-order Boltzmann function. Since intensity coding in the inner ear depends on an interaction of nonlinear basilar membrane motion and nerve fibers with three different types of synaptic threshold and growth function, the process is directly dependent on an intact mechanoelectrical transduction of outer hair cells. According to the proposed model, a loss in efficiency of outer hair cell mechanoelectrical transduction must lead to both a reduction in gain (i.e., hearing loss) and a linearizing of the response. As a result, once above threshold, the changes of stereociliary displacement, basilar membrane displacement and neural firing rate per unit change of sound intensity must be larger than for the healthy cochlea with its compressive nonlinearity. PMID- 8707630 TI - [Positron emission tomography for primary tumor detection in lymph node metastases with unknown primary tumor]. AB - In a prospective study 16 patients with cervical lymph node metastases from an occult tumor were analyzed by (18F)FDG positron emission tomography (PET). The aim of this study was to locate the primary tumor. In 7 patients a localized increase in activity was seen on PET, with this site of activity histologically proven in 4 patients. No biopsies were taken in 3 patients in areas with a high risk of complications. In a follow-up procedure at 2-22 months after primary radiation therapy in the other 9 patients, no primary tumor has been identified to date. PMID- 8707631 TI - [Diagnosis and therapy of dysphagia after partial horizontal laryngectomy]. AB - Due to an increase in the numbers of function-preserving laryngectomies and the utilization of laser techniques in cancer surgery, more patients after supraglottic partial laryngectomies are experiencing dysphagia of different intensities. We therefore developed a functional therapy program that we use for patients after supraglottic partial laryngectomies following a special diagnostic evaluation including video-laryngoscopy and, if necessary, high-speed cine radiography. This therapeutic treatment was developed from conventional treatment for patients with dysphagia caused by neurological disorders and includes body posture, altered mouth movements, food consistency and swallowing technique. In so doing, exercises are used to achieve closure of the vocal folds, aids for swallowing liquids and general directions. This method for evaluation and treatment is presented in 12 patients as examples. We now report our positive long-term results and demonstrate the possibilities and limits of therapy. Additional procedures for isolated cases are discussed critically with regard to their indications and treatment. PMID- 8707632 TI - [Manifestation of tuberous sclerosis in the ENT area]. AB - Tuberous sclerosis (Bourneville-Pringle's disease) is a rare, largely autosomal dominant neurocutaneous disease. The disease can also result from spontaneous mutations. Although strongly variable in its manifestation, manifestations are typically characterized by involvement of the central nervous system (early childhood seizures), skin (facial angiofibromas) and kidneys (angiomyolipomas). In the case described, a 67-year-old female patient complained exclusively of obstructed nasal breathing that was found to be due to angiofibromas in the nasal vestibule. Oral fibromas were asymptomatic, while fibromas in the facial region resulted in some cosmetic changes. This exclusively ENT manifestation of a patient with tuberous sclerosis has not been described previously. As treatment, the fibromas were ablated by an Nd:YAG laser under local anesthesia. Other therapeutic options are described. Additional clarification of all organ manifestations is advisable in view of numerous possible pathologies present. Genetic consultation is also recommended, particularly for patients with an oligosymptomatic variant. PMID- 8707633 TI - [Intra-orbital space occupying lesion. Differential diagnostic considerations based on a rare case]. AB - The differential diagnosis of an intraorbital tumor is discussed on the basis of a rare case of cyst formation that presented clinically in the lacrimal duct. Rapid progression of exophthalmos and visual loss required an immediate surgical intervention. Using a medial orbitotomy, a cystic tumor was extirpated that had no connection to the paranasal sinuses. Postoperatively, the visual defect and exophthalmos resolved completely. This case demonstrates the problems of differential diagnosis within the orbit that require close interdisciplinary cooperation. The final diagnosis can often be made only after surgical exploration and histopathology of tissue specimens. PMID- 8707634 TI - [Igniting the endotracheal tube in tracheotomy with electrocautery. How it should not be done]. PMID- 8707635 TI - [Sensible utilization of modern hearing aid technique. II: Multiple channel compression]. PMID- 8707636 TI - Hospitalizations for painful episodes: association with school absenteeism and academic performance in children and adolescents with sickle cell anemia. AB - Painful episodes have been identified as one of the most frequent manifestations of sickle cell anemia (hemoglobin HbSS). This retrospective study compared the frequency of hospitalization and the academic performance of two groups of children with HbSS (ages 8 to 18 years) with differing frequencies of pain. A high frequency (HF) group (n = 10) was composed of children who had four or more hospitalizations for pain in the study period; those in the low frequency (LF) group (n = 11) had one or no hospitalizations for pain during the study period. The two groups were matched on age (within 6 months), gender, and ethnicity. Standardized assessments of academic achievement and school records of attendance and class grades were obtained for all participants. The standardized academic achievement for both groups was approximately one standard deviation below the normative mean of the population sample, and class grades were below a C average. School absence was frequent in both groups (LF mean = 16.8 days/year; HF mean = 35.4 days/year), and children in the HF group had significantly more absences than children in the LF group. The lack of difference in academic performance between the two groups suggests that there may be factors other than school absenteeism that affect academic achievement, which require further investigation. PMID- 8707637 TI - Family functioning following pediatric intensive care unit hospitalization. AB - The purpose of this cross-sectional study was to compare two groups of families on measures of current family functioning and to explore factors related to current family functioning. Surveys were mailed to parents from families (n = 27) whose child aged 1 to 5 years had been hospitalized in a pediatric intensive care unit (PICU) and parents from families (n = 25) whose child aged 1 to 5 years had been hospitalized on a general care unit (GCU) in a large Midwestern children's hospital within 3 years of their child's discharge. Time since discharge ranged from 16 to 158 weeks (M = 84.9, SD = 36.0). The Cohesion and Adaptability subscales of the FACES III and the Feetham Family Functioning Survey (FFFS) were used to measure family functioning, and the Pediatric Risk of Mortality Scale (PRISM) was used to measure child acuity during hospitalization. Family functioning scores were not significantly different for PICU and GCU families. PRISM scores were negatively related to family adaptability for mothers and to family cohesion for fathers. Length of hospital stay was negatively related to fathers' adaptability ratings. Length of stay and PRISM scores were significant predictors of fathers' cohesion ratings; PRISM and location (PICU vs. GCU) were significant predictors of mothers' adaptability ratings. The findings suggest that it is not a PICU admission alone that has negative effects on the family; rather, the additive effects of how sick the child is on admission, where the child is hospitalized, and how long the child stays in the hospital may have negative consequences for the family. PMID- 8707638 TI - The sociocultural context of families with a child with myelomeningocele. AB - The purpose of this article is to describe in greater detail the sociocultural context of, and its level of supportiveness for, families with a child with myelomeningocele. Using the Family Management Style framework and a semistructured interview guide, the authors interviewed 30 mothers and fathers in this qualitative study. Families were stratified by age of the child with myelomeningocele. Fifteen categories reflecting components of the sociocultural context were identified. Across the fifteen categories, three major themes emerged: (a) Sharing Information, (b) Interpersonal Interactions, and (c) Supports/Hindrances. Levels of supportiveness across the three themes were described as supportive, moderately supportive, moderately nonsupportive, and nonsupportive. An exemplar case is presented for each. To assist families better, nurses need to assess each family's sociocultural context and its perceived level of supportiveness in the course of managing the child's chronic condition. Families require developmentally appropriate and current information specific to their situations. Interpersonal interactions should be individualized with a goal of empowering the family. Nurses should also determine supports or hindrances that may be modified through the plan of care. PMID- 8707639 TI - Comparison of morning and afternoon feedings in very low birth weight infants. AB - The purpose of this study was to compare sucking and breathing rhythmicity in very-low-birth-weight (VLBW, < 1,500 g) infants during morning feedings and afternoon feedings. Nine infants were fed at 9 a.m., and nine at 3 p.m. Eight morning-fed infants and 8 afternoon-fed infants had a continuous sucking (CS) period at the beginning of the feeding; 9 morning- and 8 afternoon-fed infants had an intermittent sucking (IS) period during which sucking bursts alternated with sucking pauses. Morning-fed infants had a lower respiratory rate during CS (p < .1) and IS (p < .05), a greater decrease in minute ventilation from the pre feed period to the CS period (p < .05), and lower sucking pressure during IS (p < .05) than afternoon-fed infants. Breathing and sucking rhythmicity scores were calculated as an indicator of the coordination of sucking and breathing. Morning fed infants had lower scores than afternoon-fed infants (p < .05). These findings suggest that morning feedings are more difficult than afternoon feedings for VLBW infants and point to the need for further studies to compare morning and afternoon feedings. PMID- 8707640 TI - Preparing staff nurses to teach parents about newborn behavior. AB - The purpose of this project was to teach postpartum nurses about the many facets of newborn behavior and to help these nurses teach parents to recognize and respond to their newborn's cues, states, and behaviors. The Keys to Caregiving: Self-Instructional Video Series was used to accomplish this goal. Eighteen maternal-child nurses from a large metropolitan health sciences center completed the program. The average pre-test score was 80%; the average post-test score was 94%. Based on the pre- and post-test scores, the results demonstrate an improvement in nurses' knowledge about newborn behavior after completing the Keys to Caregiving program. The responses from staff nurses who completed the program were overwhelmingly positive. PMID- 8707641 TI - Identification and referral by nurses of children at risk. AB - This study conducted in a pediatric hospital was initiated to determine whether an educational program focusing on the identification and referral of children at risk for developmental disturbances or variation would increase nurses' ability to identify such children and increase the actual number of referrals. Data analysis comparing pre- and post-intervention referral rates revealed fewer children being referred than should have been. The number of referrals did not differ significantly between pre- and post-education groups. Implications for policy and practice are discussed. PMID- 8707642 TI - Listen to me! PMID- 8707643 TI - The development of an instrument to measure the effects of a parent's chronic illness on parenting tasks. AB - An instrument was developed to assess the impact of chronic illness on the performance of parenting tasks (e.g., lifting, feeding, transporting, etc.). Little research on the affect of illness in general on the performance of parenting tasks is in the literature. Mothers with narcolepsy (n = 20) were used as the sample for this pilot study along with a comparison of mothers without a chronic illness (n = 20). Mothers were matched by the age and sex of the oldest child. The age groups of childhood and adolescence from the Parenting Tasks Index (PTI) were used in this study. Good reliability and validity were found in the use of this instrument. Over 50% of the mothers with narcolepsy indicated many parenting tasks (e.g., assigning the child chores, disciplining, and performing household chores) that posed moderate to severe difficulty for them. Pediatric nurses will find this instrument useful in planning individualized interventions related to identified difficult parenting tasks and in furthering research in this area. PMID- 8707644 TI - The lived experience of childhood loneliness: a phenomenological study. AB - The purpose of this phenomenological study was to describe the experience of childhood loneliness from the perspective of the child. Fourteen children aged 8 to 10 years, from middle-class White families were interviewed regarding their experiences of loneliness. Data were analyzed using Spiegelberg's (1982) method of phenomenological analysis. Thematic analysis involved a search for the experiential structures that made up the experience of loneliness. Nine themes were discovered, each presenting an aspect of the phenomenon. Unhappily Disconnected was intuited as a unity of meaning for the complex and multidimensional experience of childhood loneliness. New insights and understandings of this phenomenon may enable nurses to care more sensitively for children who are lonely. PMID- 8707645 TI - Mixed blessings: an essay. AB - It has been 12 years since Robert and Peggy Stinson chronicled the brief life of their prematurely born son in The Long Dying of Baby Andrew. Their story examined the ethical dilemmas involved in providing neonatal intensive care to marginally viable infants. Although much has changed in neonatal intensive care in 12 years time, issues regarding treatment of marginal infants remain unresolved. Further, neonatal intensive care has provided society a window through which to observe the growing fetus, with potentially profound implication for women's reproductive decision-making. The unintended and unanticipated consequences of neonatal intensive care are examined according to Roger's change model, as described in his book Diffusion of Innovations. PMID- 8707646 TI - Communication and control: parent, child, and health care professional interactions during painful procedures. AB - Through use of a qualitative ethological approach, observations of 17 children who were undergoing 44 painful procedures during cancer diagnosis or treatment were videotaped and analyzed. The children, aged 4 to 18 years, were part of a larger study testing the effectiveness of nonpharmacologic pain management techniques. Analysis of the videotaped observations revealed that several distinct patterns of conversation between caregivers, parents, and children varied greatly among situations. Both child-centered and nonchild-centered communications were demonstrated. During periods of quiet, nonchild-centered behaviors increased. As a child's distress increased, parents actively changed behaviors to redirect verbal support back to the child and to the pain control interventions. Nurses' encouraging parents to be actively involved and physically close during painful treatments may results in less distress and discomfort for the child. In addition, health care professionals need to be aware of the various patterns of child-parent-caregiver interactions and the need to stay focused on the child during painful procedures to enhance the child's ability to cope. PMID- 8707647 TI - Nurses' use of nonpharmacologic techniques with hospitalized children. AB - Pediatric nurses are often present with children who are in discomfort or pain, and are essential to successful management of this pain. Selected nonpharmacologic techniques are effective in reducing children's discomfort and pain, and thus have value in nursing practice. In a descriptive, exploratory study, 54 pediatric nurses responded to a questionnaire that elicited self-report of their use of five nonpharmacologic techniques: distraction, focusing on breathing, focusing on relaxing, imagery, and changing perceptions of painful stimuli. Content analysis indicated that (a) nurses' lack of time and heavy workload impede their use of nonpharmacologic techniques; (b) nurses' most frequent use of nonpharmacologic techniques is with children undergoing painful procedures; and (c) nurses perceive parents as helpful in implementing nonpharmacologic techniques with children. Implications for nursing practice, education, and research are discussed. PMID- 8707648 TI - Out of the hands of babes: anger, violence, and children. PMID- 8707649 TI - Childhood violence: a nursing concern. AB - This article reviews the salient features of juvenile violence as they pertain to the factors contributing to the rise of violence and the impact of violence on public safety and health care. An overview of this pediatric health concern is addressed as well as its implications for pediatric nursing care and practice. PMID- 8707650 TI - Integrative review of effects on children of witnessing domestic violence. AB - The purpose of this study was to conduct an integrative review of research studies concerning the effects on children of witnessing domestic violence. Databases were selected from MEDLINE, CINAHL, Health, and PsychInfo. Inclusion criteria required primary research in refereed journals during the last two decades that focused on children who had witnessed domestic violence (n = 14 studies). A total of 912 children and their parents comprised the subjects of this review. General agreement existed in this literature that children from families with domestic violence tended to have more difficulties than children from nonviolent families. Multidisciplinary findings revealed that the effects of witnessing violence were serious, varied, and generally framed in emotional and behavioral manifestations. Implications of these effects for clinical nursing research are considerable and merit further study. PMID- 8707651 TI - Anger disorders: basic science and practice issues. PMID- 8707652 TI - Measuring the experience, expression, and control of anger. PMID- 8707653 TI - Ideal treatment package for children and adolescents with anger disorders. PMID- 8707654 TI - The search for solutions: veterinarians explore causes of Gulf War syndrome. PMID- 8707655 TI - Right to pet ownership closer for seniors and disabled in federally assisted housing. PMID- 8707656 TI - Proper nutrition a concern for ultimate athlete. Energy expenditure and nutrition top themes in proceedings available from the International Sled Dog VMA. PMID- 8707657 TI - Opinion on possible ban on ruminant by-products in ruminant feeds. PMID- 8707658 TI - More on cost of veterinary education and perceived deficiencies of new graduates. PMID- 8707659 TI - Hiring new graduates--from a new grad's perspective. PMID- 8707660 TI - Urges reconsideration of AVMA's withdrawal from CAST. PMID- 8707661 TI - Comments on disclosure of financial interest. PMID- 8707662 TI - Domestic concern about feral cats. PMID- 8707663 TI - Comments on disclosure of financial interest. PMID- 8707664 TI - Comments on disclosure of financial interest. PMID- 8707665 TI - The best way to prevent foodborne disease and enhance the reputation of the veterinary profession. PMID- 8707666 TI - The role of food animal practitioners in control and eradication of diseases. PMID- 8707667 TI - Animal selection and breeding techniques that create diseased populations and compromise welfare. PMID- 8707668 TI - What is your diagnosis? Peritoneopericardial herniation of the liver in a cat. PMID- 8707669 TI - What is your diagnosis? Intussusception in an ostrich. PMID- 8707670 TI - Care and treatment of service dogs and their owners. PMID- 8707671 TI - Services provided by and veterinary education of small ruminant practitioners. PMID- 8707672 TI - Evaluation of outbreaks of disease attributable to eastern equine encephalitis virus in horses. AB - OBJECTIVES: To evaluate outbreaks of disease attributable to eastern equine encephalitis virus (EEEV) in horses in Michigan, and the associated environmental patterns and weather conditions, so that factors could be identified that may have predisposed horses in specific areas of the state to infections with EEEV. DESIGN: Epidemiologic retrospective records analysis. ANIMALS: Data on EEEV vectors, wild-bird reservoir hosts, and incidental hosts, including horses and human beings, obtained from census reports and medical records compiled between 1942 and 1991. PROCEDURE: Patterns detected during outbreaks of disease attributable to EEEV infections in horses were compared to associated water drainage patterns, distributions of EEEV vectors, wild-bird reservoir and incidental hosts, and weather conditions. RESULTS: Michigan has all of the elements required to sustain EEEV on a state-wide basis. Outbreaks of disease attributable to EEEV in horses have recurred in a similar regional distribution in Michigan. Regions of Michigan that have specific patterns for water drainage, specific mosquito species, and areas with higher than expected amounts of precipitation have been associated with outbreaks of disease attributable to EEEV in horses. CLINICAL IMPLICATIONS: Evaluation of environmental patterns, weather conditions, and vector and reservoir host distributions may be useful to identify areas in Michigan and elsewhere in which horses and human beings are at increased risk for an outbreak of disease attributable to EEEV. PMID- 8707673 TI - Effect of illness not associated with the thyroid gland on serum total and free thyroxine concentrations in cats. AB - OBJECTIVE: To determine circulating concentrations and fate of total and free thyroxine (T4) in cats with various illnesses not associated with the thyroid glands (nonthyroidal illnesses). DESIGN: Prospective study. ANIMALS: 98 cats with nonthyroidal illness and 50 clinically normal cats. PROCEDURE: Serum total T4 concentrations were measured by radioimmunoassay, and serum free T4 concentrations were measured by direct equilibrium dialysis. Free T4 fraction was calculated from these 2 values. RESULTS: Serum total T4 concentrations were significantly (P < 0.001) lower in sick cats (mean +/- SD, 17.18 +/- 8.14 nmol/L), compared with healthy cats (mean +/- SD, 26.00 +/- 7.62 nmol/L). Serum total T4 concentrations were inversely correlated with mortality. Differences in serum free T4 concentrations in sick cats (mean +/- SD, 27.70 +/- 13.53 pmol/L), compared with healthy cats (mean +/- SD, 24.79 +/- 8.33 pmol/L), were not significant. A few sick cats had serum free T4 concentrations greater than the reference range. Calculated free T4 fraction was significantly (P < 0.001) greater in sick cats (mean +/- SD, 0.24 +/- 0.30%), compared with healthy cats (mean +/- SD, 0.10 +/- 0.06%). CLINICAL IMPLICATIONS: Euthyroidism is maintained in sick cats, despite low serum total T4 concentrations. Measurement of serum total T4 concentrations is a valuable prognostic indicator. Serum free T4 concentrations should be used cautiously as a sole diagnostic criterion for confirmation of hyperthyroidism. PMID- 8707674 TI - Relationship between parental cardiac status in Cavalier King Charles spaniels and prevalence and severity of chronic valvular disease in offspring. AB - OBJECTIVE: To study the relationship between parental cardiac status in Cavalier King Charles Spaniels and development of chronic valvular disease (CVD) in offspring. DESIGN: Historical cohort. ANIMALS: 54 female and 53 male Cavalier King Charles Spaniel offspring. PROCEDURE: 7 sires, selected on the basis of their liability to develop CVD, were screened for clinical signs of CVD and assigned to 1 of 3 groups (late, intermediate, and early onset of CVD). The mates of these sires (30 dams) were selected and classified likewise, and 107 offspring produced in 1988 from matings between these parents were screened for clinical signs of CVD at a mean age of 5.3 +/- 0.3 years. RESULTS: 55% of the offspring were free from clinical signs of CVD, whereas 45% had cardiac murmurs of low or moderate intensity. The proportion of offspring with heart murmurs and the intensity of murmurs were significantly greater with increased parental classification. More males than females had developed murmurs, and murmurs of moderate intensity also were more prevalent in males. Results of multiple regression analysis indicated that mean parental classification and sex had significant effects on proportion of offspring with murmurs and their intensity. Additionally, age affected disease prevalence and severity, despite the narrow range in age of offspring examined. CLINICAL IMPLICATIONS: Parental CVD status is an important factor influencing the probability of heart murmurs and their intensity in offspring. The results of this study indicate that CVD development is a polygenic threshold trait and that sex of the offspring influences threshold levels. PMID- 8707676 TI - Subcutaneous accumulation of chyle after thoracic duct ligation in a dog. AB - A 3.5-year-old female German Shepherd Dog developed subcutaneous accumulation of chyle in a hind limb after ligation of the thoracic duct for treatment of chylothorax. Lymphangiography revealed severe lymphangiectasia in the abdomen and left inguinal region. Although the pleural effusion resolved, the subcutaneous accumulation of chyle continued even after medical and surgical treatment, which included transplantation of the greater omentum into the subcutaneous tissues of the affected limb. Alternate lymphaticovenous pathways usually develop in dogs after thoracic duct ligation. These apparently did not develop in this dog, resulting in retrograde flow of chylous fluid into the hind limb. To the authors' knowledge, this is a previously unrecognized complication of thoracic duct ligation for treatment of chylothorax in dogs. PMID- 8707675 TI - Effect of intravenous administration of fluids on packed cell volume, blood pressure, and total protein and blood glucose concentrations in healthy halothane anesthetized dogs. AB - OBJECTIVE: To determine the effects of IV administration of fluids on PCV, serum total protein and blood glucose concentrations, and systolic arterial pressure in healthy anesthetized dogs undergoing elective surgical procedures. DESIGN: Prospective, randomized controlled trial. ANIMALS: 70 clinically normal dogs. PROCEDURE: Dogs received i.v. administration of 0, 5, 10, or 15 mL/kg of body weight/h of a polyionic crystalloid solution or 5% dextrose in water. Blood samples were collected before and after administration of medication, prior to anesthetic induction, after anesthetic induction, at the end of the surgical procedure, and 2 hours after surgery to determine PCV and serum total protein and blood glucose concentrations. Blood pressure was measured before and after anesthetic induction and at the end of the surgery. RESULTS: There were not any significant differences in PCV, total protein concentration, or systolic arterial pressure among treatment groups. Hyperglycemia developed in dogs receiving 5% dextrose in water, but resolved 2 hours after discontinuing administration of fluids. CLINICAL IMPLICATIONS: Intravenous administration of fluids may not be necessary to maintain normal blood pressure in young, healthy dogs undergoing elective surgery. PMID- 8707677 TI - Unilateral nephrectomy in dogs with renal disease: 30 cases (1985-1994). AB - OBJECTIVE: To evaluate indications for and complications, efficacy, and effects on renal function of unilateral nephrectomy in dogs with renal disease, and to evaluate the role that scintigraphy had in the decision to excise a kidney. DESIGN: Retrospective case series. ANIMALS: 30 dogs with renal disease that underwent unilateral nephrectomy. A comparison group of 12 dogs with renal calculi that underwent renal scintigraphy but not nephrectomy was included. RESULTS: Indications for nephrectomy included renal or ureteral calculi (n = 10), renal mass (8), chronic pyelonephritis (5), perirenal mass (3), severe hydronephrosis and hydroureter (3), and renal hypoplasia with ureteral ectopia (1). None of the dogs were azotemic before surgery. Renal scintigraphy apparently influenced the decision to perform nephrectomy, because in 14 of 16 dogs that underwent nephrectomy, the affected kidney contributed < or = 33% of the total glomerular filtration rate, but in 6 of 8 comparison dogs that underwent nephrotomy, the affected kidney contributed > 33% of total glomerular filtration rate. Complications of nephrectomy included oliguria (5) and organ laceration (2). Mean +/- SD final serum creatinine concentration for 16 dogs alive at least 6 months after nephrectomy was 2.2 +/- 1.8 mg/dl. Three dogs had chronic renal failure of undetermined cause at the time of death. Nephrectomy did not completely resolve the underlying disease in 13 dogs. Renal function was evaluated in 6 dogs 2 to 3.5 years after nephrectomy and was impaired in 4. None of the dogs were anemic, azotemic, proteinuric, or hypertensive. Survival time varied depending on the underlying disease. CLINICAL IMPLICATIONS: Multiple factors contributed to the decision to perform nephrectomy. Unilateral nephrectomy resulted in few serious complications and was not detrimental to the remaining kidney, but did not always resolve the underlying disease. PMID- 8707678 TI - Hormonal indicators of pregnancy in llamas and alpacas. AB - OBJECTIVE: To determine concentrations of estrone sulfate in serum, estrone sulfate in urine, relaxin in serum, and progesterone in serum in pregnant llamas and alpacas and to assess the potential of these hormones as pregnancy indicators. DESIGN: Prospective study. ANIMALS: 19 parous pregnant camelids (8 llamas and 11 alpacas). PROCEDURE: Estrone sulfate concentrations (in serum and in urine) and progesterone concentrations (in serum) were determined by enzyme immunoassay. Relaxin concentrations (in serum) were measured by radioimmunoassay. Serum and urine samples were collected daily for the first 30 days after breeding and, thereafter, once weekly until parturition. RESULTS: Estrone sulfate concentrations (in serum and in urine) peaked twice during pregnancy. The first took place 21 days after breeding and the second during the last month of pregnancy. Relaxin concentrations increased at 3 months of gestation to > 20 ng/mL, decreased at 5 months to 5 ng/mL, then increased from 8 months of pregnancy until parturition. Progesterone concentrations were detectable 4 days after breeding and were maintained > 2 ng/mL throughout pregnancy. CLINICAL IMPLICATIONS: The first increase in estrone sulfate concentration over basal values may indicate early interaction between mother and embryo, whereas the second increase may reflect fetal viability. Use of estrone sulfate concentration to diagnose pregnancy in llamas and alpacas is highly dependent on time of sampling. Relaxin concentration in serum is a superior indicator of pregnancy after the second month in the Ilama and alpaca because its existence is suggestive of interaction between mother and fetus, and concentrations are greater than basal values for a long period of pregnancy. Progesterone is not a direct product of the embryo or fetus and only indirectly confirms a diagnosis of pregnancy. PMID- 8707679 TI - Efficacy of a temperature-sensitive modified-live bovine herpesvirus type-1 vaccine against abortion and stillbirth in pregnant heifers. AB - OBJECTIVE: To evaluate the efficacy of a commercially available temperature sensitive modified-live bovine herpesvirus type-1 (BHV-1) vaccine against BHV-1 challenge-induced abortion and stillbirth. DESIGN: Prospective randomized control trial. ANIMALS: 20 cycling, nonpregnant, BHV-1 seronegative heifers of various breeds and weights, 12 to 15 months old. PROCEDURE: Heifers were randomly assigned to a vaccinate (n = 10) or nonvaccinate control (n = 10) group. Seventeen to 26 days after members of the vaccinate group received a second dose of vaccine, all heifers were artificially inseminated. Heifers were challenged intravenously with Cooper strain BHV-1 between days 177 and 187 of gestation. Aborted fetuses and stillborn calves were necropsied, and tissues collected for histologic examination and virus isolation. Heifers, calves, and fetuses were tested for BHV-1 antibody throughout the study. RESULTS: The difference in number of abortions or stillbirths between vaccinated heifers (1/10) and control heifers (10/10) was significant (P < 0.003). Seven of 10 control heifers had a virus neutralization antibody titer to BHV-1 at abortion or stillbirth that declined or remained unchanged from their titer at a previous serologic evaluation (7 to 66 days earlier). CLINICAL IMPLICATIONS: Prebreeding vaccination of replacement heifers with modified-live BHV-1 vaccine provides fetal protection at 6 months of gestation (7 months after vaccination) and appears to be a reasonable precaution to control economic losses associated with BHV-1 infection. Abortions induced by BHV-1 are not necessarily associated with rising or markedly high virus neutralization antibody titers. These titers should be used cautiously when assessing the role of BHV-1 in bovine abortion and stillbirth. PMID- 8707680 TI - Epidemiologic and pathologic characteristics of respiratory tract disease in dairy heifers during the first three months of life. AB - OBJECTIVE: To describe the incidence of respiratory tract disease in dairy calves and to compare antibody titers and microbial isolates from transtracheal wash samples between calves with and without respiratory tract disease (cases and controls, respectively). DESIGN: Prospective observational cohort study, with matched case-control substudy. ANIMALS: 410 dairy heifers; in substudy, 105 cases and 59 controls from the same population. PROCEDURE: Calves were examined weekly by a veterinarian during the first 3 months of life. Blood samples were collected for serologic testing at the first visit for each calf and during acute and convalescent periods for cases. Transtracheal wash samples also were obtained during the acute period from cases and controls. RESULTS: Incidence and case fatality risk for clinician-diagnosed pneumonia were 25.6 and 2.2%, respectively. Mycoplasma sp and Pasteurella multocida together were isolated from 29% of cases and 11% of controls, and Mycoplasma sp alone from 7% of cases and 30% of controls (both P < or = 0.05). From postcolostral to acute-phase serum samples. Mycoplasma dispar titers increased 1.3-fold among cases, compared with 0.7-fold among controls; from acute- to convalescent-phase samples, M dispar titers increased 2.4-fold among cases, compared with 5.6-fold among controls (both P < or = 0.005). CLINICAL IMPLICATIONS: Results of this study suggested a synergistic effect between Mycoplasma sp and P multocida and a possible initiative role of M dispar in the development of respiratory tract disease. Postcolostral total IgG values and antibody titers were not significantly different between cases and controls, implying that other factors have an important role in the development of respiratory tract disease. PMID- 8707681 TI - Morbidity from nonrespiratory diseases and mortality in dairy heifers during the first three months of life. AB - OBJECTIVE: To describe causes of death, mortality, and morbidity from nonrespiratory diseases in dairy calves. DESIGN: Prospective observational cohort study. ANIMALS: Convenience sample of 410 dairy heifers born between January and December 1990 in 18 south-western New York herds. PROCEDURE: Heifers were examined weekly by a veterinary clinician during the first 3 months of life and all disease conditions were recorded. RESULTS: Crude risks for diarrhea, umbilical infection, and umbilical hernia were 28.8, 14.2, and 15.1%, and the median ages at first diagnoses were 2, 1, and 3 weeks, respectively. Mean durations of umbilical infection and umbilical hernia were 3.7 and 6.7 weeks, respectively. Crude mortality was 5.6%. Case-fatality risks were 12.8% for diarrhea during the first week of life, 5.1% for diarrhea after the first week of life, and 0% for umbilical infection and umbilical hernia. Diarrhea was diagnosed by the caretaker of the clinician; umbilical conditions were diagnosed by the clinician. The primary cause of death was diarrhea in 43%, pneumonia in 24%, septicemia in 10%, and other single causes in the rest of the 21 necropsied calves. CLINICAL IMPLICATIONS: The high incidence and somewhat long duration of umbilical infection, the finding that diarrhea was the primary cause of death, and the high case-fatality risk for diarrhea during the first week of life suggested that calf caretakers need training in the prevention and treatment of these conditions. PMID- 8707682 TI - Prediction of calf mortality by use of tests for passive transfer of colostral immunoglobulin. AB - OBJECTIVE: To examine the ability of several commonly used tests for evaluation of passive transfer of immunoglobulin to predict mortality in dairy replacement heifers. DESIGN: Prospective observational study. ANIMALS: 246 dairy replacement heifers between 1 and 8 days of age. PROCEDURE: Using serum samples obtained from each calf, total serum protein concentration and results of zinc sulfate turbidity, sodium sulfite turbidity, radial immunodiffusion, and glutaraldehyde coagulation were determined. Calves were monitored for 100 days, and relative risks for death were calculated. Logistic regression models predicting death also were developed. RESULTS: None of the logistic regression models detected a significant association between test results and mortality. The greatest relative risks of mortality were observed in calves with serum protein concentrations < 4.5 g/dl, serum IgG1 concentrations < 500 mg/dl, and sodium sulfite test scores < 1+. CLINICAL IMPLICATIONS: Calves with lower passive transfer values had increased risk of death; however, failure of passive transfer is not an infallible predictor of mortality. PMID- 8707683 TI - Grounded theory analysis of car drowning and women's health. PMID- 8707684 TI - Graduate nursing concentration in women's health at the University of Illinois at Chicago. AB - The Graduate Nursing Concentration in Women's Health was formalized in the 1980s as a program of study that emphasizes nursing theory, research, and practice related to women's health. We describe the purposes of the program and the assumptions underlying the care of women as well as the curriculum and characteristics desired of graduates. In this program, the goal of advancing women's health is balanced by the desire to create a productive, educational environment that facilitates the personal construction of knowledge by learners. PMID- 8707685 TI - Multiple roles of women and role strains. AB - Women are likely to experience multiple roles, often several at the same time, for which different sets of responsibilities are designated. Expectations that others have of us in any one of our roles may differ from our own, and conflict with our goals as individuals. The multiplicity and overlapping of roles often provoke conflict and stress. Depending on a number of variables, growth of the individual is either impeded or promoted. Vignettes of women experiencing overlapping roles, at various stages of the life cycle, are cited to illustrate the complexities of the issues and coping styles. PMID- 8707686 TI - Problems women patients experience in the medical encounter for chronic pelvic pain: a New Zealand study. AB - Women who have used the New Zealand health services for chronic pelvic pain have reported problems with diagnosis, communication with the doctor, lack of information, and inappropriate treatment. Six focus groups were conducted in which women talked about their experiences of using the health services for chronic pelvic pain. A discourse analysis of this material provided a conceptual basis for understanding the key dimensions structuring women's experience. Themes of particular concern were that (a) doctors negated the woman's own experience of the pain and its meaning to her and this negation affected her ability to take responsibility for her condition; (b) the medical "extraction" of symptoms through technology overshadows the woman's discursive interpretation of, and voicing of, her symptoms; and (c) the concept of "pelvic pain without organic pathology" implies an absence that becomes filled with the specter of neurosis or psychological problems. PMID- 8707687 TI - Problems of communication, diagnosis, and treatment experienced by women using the New Zealand health services for chronic pelvic pain: a quantitative analysis. AB - Women who have used the New Zealand medical services for chronic pelvic pain report problems with diagnosis, communication with the doctor, lack of information, and inappropriate treatment. There is a sense of increasing loss of control as repeated visits to the doctor do not produce an understanding of the pain. I conducted a survey to investigate the extent to which these problems are affecting the total group of women using the health services for pelvic pain in New Zealand and to ascertain if any particular group of women is more likely to be experiencing such problems. The pattern of responses showed that the problematic facets of the medical encounter formed an interactive pattern: Women who had one type of problem tended to have others. Between one fourth and one third of the sample experienced these problems. Problems surrounding communication with the doctor, diagnosis, medication, and surgery are discussed. Women of lower socioeconomic status were found to be more likely to experience these problems. PMID- 8707688 TI - Safe child care and women's empowerment in the developing world. AB - Women's ability to provide for the health and well-being of their children and families is linked in complex ways to the degree of empowerment they experience. Empowerment for women in the developing world is a perplexing venture, hampered by patriarchal barriers and cultural confusion. Our purpose in conducting this research was to discover how disadvantaged Namibian women experienced empowerment when they had access to safe child care. Fifty-two women participated in the study. A qualitative design and ethnographic techniques were used to elicit women's experiences. Five themes, descriptive of women's empowerment, emerged from the data: (a) increasing vitality, (b) freedom from worry, (c) opportunities to increase financial security, (d) strengthened parenting competence, and (e) personal satisfaction and a sense of fulfillment. The findings add a voice from the developing world to the growing understanding of women's diverse journeys toward empowerment. Discussion focuses on the significance of the themes and implications for further research that promotes women's empowerment and community health. PMID- 8707689 TI - Premenstrual syndrome in employed Chinese women in Hong Kong. AB - Premenstrual syndrome (PMS) has been studied in many countries, but few studies have been reported internationally on the experience of Chinese women. Because culture and employment are important factors in the perception of health status, in this study we sought to determine the existence and features of PMS in Chinese clerical women in Hong Kong. We used a cross-sectional, retrospective approach to collect data with a translation of an established questionnaire (the Menstrual Distress Questionnaire). Fatigue was found to be the most prevalent symptom, and the Pain, Water Retention, Behavioral Change, and Negative Affect scales had more than 64% frequency. The main difference between these findings and those of other studies is that negative affect featured most prominently in Western samples, whereas pain featured most highly in this sample of Chinese women. PMID- 8707690 TI - Health effects of experiences of sexual violence for women with abusive partners. AB - We assessed the incidence of sexual violence, physical violence, physical health symptoms, gynecological symptoms, and risk behaviors for contracting an STD or HIV infection in women who had used a shelter for women with abusive partners. In addition, we investigated the relationships between sexual violence and the frequency of physical health symptoms, including specific gynecological symptoms. Results indicated that one fourth of the women interviewed had experienced sexual violence and nearly two thirds of the women had experienced physical violence in the past 6 months. The incidence of physical health symptoms, gynecological symptoms, and risk behaviors for exposure to STDs and HIV infection are presented. The correlations among sexual violence, physical violence, and experiences of physical health symptoms are also reported. This study is particularly valuable because previous research has not documented the relationship between sexual violence and physical health symptoms. PMID- 8707691 TI - Impact of HIV/AIDS on women in the United States: challenges of primary and secondary prevention. AB - Women are the fastest growing group in the United States being infected with HIV. Although the majority of people with AIDS in the United States are men, AIDS is increasing almost four times as fast among women. Yet women remain understudied, underreport, and underdiagnosed. There are insufficient resources and services targeting the primary prevention needs of women, so women keep getting infected. Secondary prevention efforts for women infected with HIV have been stymied, and nearly a decade has been lost that might have been devoted to early detection of HIV infection in women, documentation of the natural history of HIV in women, exploration of their health needs, expansion of gender-specific services, and development of interventions to improve quality and length of life for HIV positive women. In this article, I analyze the impact of HIV/AIDS on women in the United States and pose challenges for primary and secondary prevention. PMID- 8707692 TI - The future of Social Security and its impact on older women. PMID- 8707693 TI - Is Rubin's "taking-in" and "taking-hold" a useful paradigm? AB - Rubin's "taking-in" and "taking-hold" model has been a dominant paradigm for postpartum nursing. To determine whether "taking-in" and "taking-hold" apply to contemporary hospitalized postpartum women, I conducted a repeated measures survey of 95 women with uncomplicated postpartum courses after vaginal births during their hospital stay. "Taking-in" and "taking-hold" were measured with the Martell Postpartum Questionnaire-Revised (MPQ-R), a paper-and-pencil instrument. An expert panel established the content validity of the MPQ-R in this study. Cronbach's alpha for the two subscales ranged from .40 to .60, a result of skewed distribution of items or lack of variability in scores. The participants were neither "taking-in" nor "taking-hold." The distribution of the participants' responses was similar to each others' and indicated that their postpartum experiences included some aspects of both "taking-in" and "taking-hold." Despite the limitations of the MPQ-R and the research methods, the findings suggest that Rubin's theory has limited usefulness for postpartum nursing of contemporary women. PMID- 8707694 TI - Pregnant women at risk: an evaluation of the health status of refugee women in Buffalo, New York. AB - Each year, conflicts escalate the need for women, children, and men to seek refuge outside their home countries, and the United States and Canada provide asylum for many of these refugees. However, few researchers have investigated the health status of specific populations of refugees sojourning or newly arrived in the United States. As nurses at a Buffalo, New York, refugee shelter for Canada bound asylum seekers, we provided health assessments to this group of refugees at their on-site clinic. Pregnant women were of major concern, being potentially at risk for poor pregnancy outcomes. We found that our sample of pregnant refugees had several previously undetected health problems and suggest that further research on refugee populations is needed. PMID- 8707695 TI - Delay in prenatal care as a result of battering in pregnancy: cross-cultural implications. AB - White American, Hispanic, and African American women were surveyed in public health and low-income clinics in Los Angeles, California, and Seattle, Washington, to determine if they delayed seeking prenatal care because of battering during their pregnancies. Nursing staff in the clinics attempted to enroll in the study all pregnant women from these groups who presented themselves for care; no other criteria were used, other than the ability to read either English or Spanish. Results were obtained from 162 White Americans, 208 Hispanics, and 132 African Americans. Although the incidence of abuse was not significantly different among the ethnic groups, battered women sought prenatal care 6.5 weeks later than the nonabused sample, with a similar delay in each ethnic group. Twenty-one percent of the women reported physical harm during the pregnancy, and 13.7% stated that they had delayed care because of injuries. PMID- 8707696 TI - Patterns of postpartum health in mothers of low birth weight infants. AB - Our purpose in conducting this prospective, descriptive study was to examine patterns of maternal health in women who delivered a low birth weight (LBW; < or = 2,500 g) infant until 6 months past the mothers' estimated date of confinement (EDC). Sixty-eight mothers of LBW infants were interviewed at nine data collection points: delivery, original EDC, infant discharge, and every month thereafter for 6 months. Health was assessed using the Health Perceptions Questionnaire and a structured interview (the Health Interview Schedule). Maternal acute care visits occurred throughout the study, with at least 71% of the mothers reporting one acute care visit. Sixty-four percent of the mothers reported having to change their activities because of ill health between the time their infant went home and the 6-month data point. At the 6-month data point, 19% of the mothers still rated their health as fair to very poor. An ongoing assessment of maternal health is important in mothers of LBW infants so that their health can be optimized. PMID- 8707697 TI - The impact of a history of rape and incest on the posthysterectomy experience. AB - I conducted a cross-sectional study to examine the impact of a history of rape and/or incest on the posthysterectomy experience. The sample consisted of 92 women recovering from elective hysterectomies. They were interviewed at 6, 12, 18, and 24 months after their operations. The Derogotis Sexual Functioning Index, the Center for Epidemiologic Studies-Depression Scale, and two hysterectomy related questionnaires were administered. An analysis of variance compared abused with nonabused women on their sexual functioning, depression level, and satisfaction with their hysterectomy decision. Abused women had a significantly (p < .05) higher level of depression in the first year after the hysterectomy but not in the second year after the operation. Results suggest the need for expanded preoperative sexual history assessments and interventions and also to define areas for further research. PMID- 8707698 TI - Determining the adequacy of a health promotion self-care interview guide with healthy, middle-aged, Mexican American women: a pilot study. AB - The Hispanic/Latino population is the second largest and fastest growing minority in the United States, with Mexican Americans comprising 60.4% of the total. Existing research instruments and procedures must be made culturally specific, both theoretically and empirically, for use with this population. Our purpose in the present study was to determine whether one part of the Health Promotion Self Care Interview Guide (HPSCIG), developed to identify self-care actions that promote well-being in healthy, middle-aged Anglo American and African American women (Hartweg, 1991, 1993), was culturally sensitive for use with Mexican American women. This was a necessary step before the full HPSCIG was translated into Spanish for a larger bilingual study. Twenty Mexican American women identified 232 self-care actions they took to promote well-being as they defined it. More than 81% of the self-care actions were performed to meet the universal self-care requisites, 11% were performed to meet the developmental requisites of middle-age, and 2% were performed to meet health deviation self-care requisites. Six percent of the actions could not be classified using the theoretical coding scheme. Inherent in the study were questions about the congruence and relevance of self-care as a concept in this population. The findings support the use of the HPSCIG with healthy, middle-aged Mexican American women. PMID- 8707699 TI - Images and sensations of war: a common theme in the history of military nursing. AB - We analyzed interviews with 22 military nurses who had served in wars or conflicts since World War II, up to and including Operation Desert Storm. Using a method of hermeneutic phenomenology, we found several common themes in the interviews. In this article, we report on the theme Images and Sensations of War. Within that theme, the subcategories sights, sounds, and other sensations (climate and weather, taste, smell, and comfort) were discovered. We categorized the images and sensations as pleasant, unpleasant, and horrible. The findings from this analysis may be useful to professionals who recruit military nurses, provide ongoing military training, help military nurses return to civilian life, and treat posttraumatic stress syndrome. Furthermore, the findings could be used as a basis for future quantitative studies to determine how widespread this theme is in larger groups of military nurses. PMID- 8707700 TI - Augmentation mammoplasty: a complex choice. AB - Augmentation mammoplasty is a surgical procedure with known risks and complications. Despite this, millions of women have opted to undergo the procedure. With the recent publicity surrounding this procedure, and the current concern about long-term side effects, the rationale behind women's choice has been questioned. Often, health care professionals are less than empathetic when a women complains about experiencing complications. This attitude seems to "blame the victim." We examine the complex psychological and sociological factors behind a woman's decision to have her breasts enlarged artificially. It is our hope that the information presented herein will contribute to a better understanding and more compassionate care of women who have breast augmentation surgery. PMID- 8707701 TI - Tony J. Cunha, 1916-1992: a brief biography. PMID- 8707702 TI - Prediction of daily protein accretion rates of pigs from estimates of fat-free lean gain between 20 and 120 kilograms live weight. AB - The objective of this study was to evaluate a method for predicting daily protein accretion rates of various genotypes of pigs reared in different environmental conditions using easily obtained mean daily fat-free growth rates. Data were obtained for seven genotype-environment groups of gilts and nine groups of barrows. Daily empty body protein accretion rates were estimated at 1.0-kg intervals between 20 and 120 kg live weight. The estimates were fitted to a generalized exponential function, PA = A e(B x WT + C/WT + D x WT2), where WT is kilograms of live weight and A, B, C, and D are estimated parameters for each sex. Nonlinear least squares methods were used to estimate the intercept and regression coefficients expressing each parameter estimate (A, B, C, and D) as a linear function of the mean fat-free lean gain for each sex-genotype-environment group. The mean percentage absolute errors were 3.5% for gilts and 6.1% for barrows. The largest errors occurred between 110 and 120 kg live weight. From 20 to 110 kg, mean percentage errors averaged 2.7% for gilts vs 4.8% for barrows. These results offer encouraging evidence that a generalized equation can be used to predict daily protein accretion rates from mean fat-free lean growth data. Further research, with additional genotype-environment populations, is needed to increase accuracy of the generalized growth functions. PMID- 8707703 TI - Effects of dietary energy source and level on performance of newly arrived feedlot calves. AB - Three experiments were conducted with Simmental x Angus crossbred steers to determine the effects of energy source and level on performance of newly arrived feedlot calves. In trial 1, 68 steers (initial BW 215 +/- 9.4 kg) were used in a 2 x 3 factorial experiment to determine the effects of receiving diet and previous creep feed on performance. Diets were composed primarily of either corn silage, corn silage + alfalfa pellets + dry corn, or dry corn + alfalfa pellets. For the 41-d trial, calves fed the corn silage-based diet had greater (P < .05) ADG and feed efficiencies than calves fed the other two diets. In Trial 2, 60 steers (initial BW 212 +/- 4.6 kg) were used in a 28-d completely randomized design experiment to determine the effects of 16% CP receiving diets containing 70, 75, 80 or 85% concentrate on performance. There were no differences (P > .10) in ADG or feed efficiency due to dietary concentrate level. In Trial 3, 77 steers (initial BW 226.3 +/- 3.0 kg) were used in a 2 x 3 factorial experiment to determine the effects of dietary concentrate and protein levels on performance during a 28-d receiving period. The factors were concentrate level (70 vs 85% concentrate) and protein level (12.5%, 16%, or phase-fed at 23%, 17%, 14%, and 12.5% during wk 1, 2, 3, and 4, respectively). During wk 1, calves fed the 85% concentrate diet had greater (P < .01) DMI, ADG, and feed efficiency compared with 70% concentrate. Calves fed the 16% CP and phase-fed protein diets had greater (P < .01) DMI, ADG, and greater (P < .02) feed efficiency than calves fed the 12.5% CP diets. Receiving diets containing at least 16% CP and > 70% concentrates are beneficial to calves during the first week after feedlot arrival. PMID- 8707704 TI - Methionine requirement of channel catfish fed soybean meal-corn-based diets. AB - A soybean meal-corn-based diet was used to determine dietary methionine (Met) required by 14-g channel catfish (Ictalurus punctatus) in a 42-d experiment at 25 degrees C. The basal diet with balanced limiting amino acids relative to the catfish whole-body amino acid profile contained 277 g of CP, 3.6 g of Met, 4.0 g of cystine (Cys), and 10 MJ of DE/kg of DM. DL-Methionine was added to the basal diet from 0 to 12.0 g/kg in 2-g intervals at the expense of L-glutamic acid to produce seven isonitrogenous and isocaloric diets. A reference diet contained 331 g of CP, 8 g of Met, 5 g of Cys, and 10 MJ of DE/kg of DM (included 8% fish meal). Seven graded Met levels resulted in quadratic responses (P < .01) of weight gain, specific growth rate, feed or GE intake, feed or energy efficiency, protein or energy retention, protein efficiency ratio, and apparent net protein or energy utilization. Channel catfish required 9.4 g of Met/kg of DM (34.1 g/kg of CP) with a total 11.3 g/kg of calculated digestible sulfur-containing amino acids based on multiple regression dose-response models or 270 mg of Met/kg of fish per day based on a broken-line response of protein gain to Met intake. At the adequate Met level, catfish with the lowest (P < .05) liver lipids showed feed intake and protein or energy utilization efficiency similar (P > .05) to that of catfish fed the reference diet. Catfish fed all-plant-protein diets require more dietary methionine than previously reported. Catfish fed corn soybean meal diets fortified adequately with methionine result in performance that approaches that of fish fed a fish meal-based diet. PMID- 8707705 TI - Genetic trend and environmental effects in a population of cattle selected for twinning. AB - A selection experiment was established in 1981 to increase twinning rate in cattle. Results reported are through 1993 calf crops. Estimates of genetic parameters for a two-trait twinning and ovulation rate model with genetic groups were as follows: heritabilities of .03 for twinning and .07 for ovulation rates with a genetic correlation of nearly 1.00 and fractional permanent environmental variances of .06 for twinning and .05 for ovulation rate. Corresponding estimates when group effects were ignored were as follows: heritabilities of .08 and .08 and fractional permanent environmental variances of .02 and .04 for twinning and ovulation rates, respectively. Twinning rate (percentage) in the project at the U.S. Meat Animal Research Center has increased in all cows born in the project by year of calving from 3.4% in 1982 to 28.5% in 1993, a phenotypic increase of 25.1%. The estimated genetic change in twinning of cows by year of calving using the groups model has been 15.2%. The increase in average genetic value by year of birth has been 18.2% in twinning and 15.0% in ovulation rate from 1980 through 1991. Solutions for seven selected groups of foundation animals ranged from -6.0 to 33.1% and influenced genetic trend. PMID- 8707706 TI - Estimates of heritabilities and genetic and environmental correlations for left- and right-side uterine capacity and ovulation rate in mice. AB - Heritabilities for and genetic and environmental correlations between uterine capacity, ovulation rate, and body mass (BM) were estimated in mice. Uterine capacity was defined as the number of fetuses (LUC or RUC for left or right side) in one uterine horn for unilaterally ovariectomized females. Ovulation rate (corpora lutea, LCL or RCL for left or right ovary) was measured on the remaining single ovary in these same females. Data on 1,931 mice from four selection populations were used. Left ovulation rate and LUC were measured on 958 animals, and RCL and RUC of another 972 animals were recorded. Genetic and environmental variances and covariances were estimated simultaneously using an animal model with a multiple-trait, derivative-free, restricted maximum-likelihood procedure. Averages for heritability and correlation estimates derived from separate analyses of the selection populations are presented below. Heritability of LUC was higher (.33 +/- .06) than that of RUC (.19 +/- .02). Heritability of LCL and RCL ranged from .17 +/- .03 to .27 +/- .06, and heritability for BM was .65 +/- .05. The genetic correlation between LUC or RUC and LCL or RCL ranged between .43 +/- .29 and .68 +/- .05, and between LUC and RUC was .92 +/- .05. Body mass had a higher genetic correlation with LCL and RCL (.70 +/- .12 and .93 +/- .02) than with LUC and RUC (.37 +/- .05 and .47 +/- .12). Environmental correlations between LCL and LUC and RCL and RUC were .32 +/- .09 and .36 +/- .05, respectively. PMID- 8707707 TI - Bad ethics, good ethics and the genetic engineering of animals in agriculture. AB - Genetic engineers have been remiss in addressing ethical and social issues emerging from this powerful new technology, a technology whose, implications for agriculture are profound. As a consequence of this failure, society has been uneasy about genetic engineering of animals and has had difficulty distinguishing between genuine and spurious ethical issues the technology occasions. Many of the most prominent concerns do not require a serious response. On the other hand, concerns about a variety of possible risks arising from genetic engineering of animals require careful consideration and dialogue with the public. Such concerns are an admixture of ethics and prudence. A purely ethical challenge, however, hitherto not addressed, is represented by problems of animal welfare that arise out of genetically engineering agricultural animals. A principle of "conservation of welfare" is suggested as a plausible moral rule to guide such genetic engineering. PMID- 8707708 TI - Repeated matings with individual ewes by rams differing in sexual performance. AB - Twenty-four sexually mature rams were individually exposed to 10 estrous ewes on three occasions to determine the relationship between levels of sexual performance (ejaculation rate or serving capacity) and the incidence of repeated matings (ejaculations) with individual ewes. Rams that attained six ejaculations in a relatively short period of time (HP rams) mated fewer times with individual ewes than low-performing (LP) rams (P < .01). As a consequence, HP rams tended to mate with a greater number of different ewes than did LP rams (P < .06). In addition, time required to attain six ejaculations was inversely correlated with the number of different ewes mated (P < .03). The HP rams did not differ from LP rams in the number of different ewes investigated and courted. These findings highlight the value of using rams with relatively rapid ejaculation rates when breeding large groups of naturally cycling or synchronized ewes. PMID- 8707709 TI - Animal performance and fleece characteristics of Angora goats maintained on western and southern Texas rangeland. AB - Two hundred castrated male Angora goats (18 mo of age) were divided equally between rangeland sites in the oak savannah of the Edwards Plateau (EP) and the mixed-brush shrublands of the South Texas Plains (STP) for 5 yr to study influence of environment and age on body weight (BW), mohair production, and mohair fiber characteristics. Goats were grazed on rangeland typical of the respective regions and were given typical management that included shearing in February and July or August of each year. Shorn goats were weighed and individual fleeces were measured for grease weight (GFW), clean yield (CMFP), average fiber diameter (AFD), and proportion of medullated fibers, med (MFP) and kemp (KFP). Rainfall and maximum and minimum temperatures were recorded daily at each site. The AFD (P = .01) and its associated SD (P < .001) and kemp levels (P = .03) were all higher at the STP than at the EP site (39.4 vs 37.9 microns, 10.4 vs 11.1 microns, and .80 vs .68%, respectively). Angora goats at EP weighed less (38.1 vs 41.7 kg, P = .03), and sheared less (1.90 vs 2.22 kg, P = .01), finer (36.4 vs 39.3 microns, P < .001), and less kempy (.52 vs .84%, P < .001) mohair at the February shearing than at the August shearing. No seasonal differences in body weight or in mohair production or quality were apparent at STP. The proportions of total variation in grease mohair production (30.6%), CMFP (22.2%), and AFD (15.7%) due to year effects were two to three times greater at STP than at EP. In contrast, year effects were responsible for only a small proportion (2.2 to 4.6%) of the variation in medullated fibers. The effects of rainfall accumulation within specific 6-mo growing periods were positive for BW, GFW, CFW, AFD, SD of AFD and MFP and negative for CMFP and KFP. Only the rainfall effects on SD of AFD and medullated fibers were significant (P < .05). The effects of age were positive for BW (P < .001), CMFP (P = .046), AFD (P < .001), MFP (P = .014), and KFP (P = .084) and negative for GFW (P = .046) and CFW (P = .107). This study documents influence of environment and age of goats on body weight, mohair production, and fiber characteristics. PMID- 8707710 TI - Beta-adrenergic receptors in porcine adipocyte membranes: modification by animal age, depot site, and dietary protein deficiency. AB - Adipocyte lipid metabolism is primarily regulated by insulin and the catecholamines norepinephrine and epinephrine. Stimulation of the beta-adrenergic receptors (beta-AR) by catecholamines causes an increase in the rates of adipocyte lipid degradation and a decrease in the rates of lipid synthesis. These catabolic effects are in opposition to insulin, which causes net anabolic effects. Because most of the postnatal development of adipose tissue mass in pigs results from hypertrophy of adipocytes (rapid in first few weeks of life) caused by increased net synthesis of triacylglycerol, there is interest in the modulation of beta-AR in adipocytes of growing pigs. The beta-AR are characterized by measuring ligand binding to the receptor to ascertain the affinity of the ligand for the receptor and the receptor number. We found the affinity of the receptor did not vary with animal age (10, 28, and 75 d), with adipose tissue depot site, or in adipocytes of protein-deficient pigs. The beta AR in obese pigs tended to have greater affinity than those in crossbred pigs of the same age and weight. The beta-AR number was not different when expressed per milligram of adipocyte membrane protein in pigs of different age, in obesity, in different adipose tissue depots, or during protein deficiency. The number expressed per cell or per unit adipocyte surface area did not differ between depots or during protein deficiency. The number per cell tended to be greater in the larger cells from 75-d-old pigs than in the smaller cells from 10- and 28-d pigs. It was greatest in obese pigs with the largest adipocytes. Under the various experimental conditions (age, obesity, depot, protein deficiency), the membrane fatty acid composition was greatly different, but in most cases there was no modulation of beta-AR affinity. PMID- 8707711 TI - Chronic effects of somatotropin treatment on response of subcutaneous adipose tissue lipolysis to acutely acting factors in vivo and in vitro. AB - The basis of the chronic lipolytic effect of somatotropin on adipose tissue was investigated in sheep. Lipolytic rate was assessed in subcutaneous adipose tissue both in vivo, by microdialysis, and in vitro. Somatotropin treatment resulted in a small increase in basal (unstimulated) lipolysis and also in the maximum lipolytic rate observed in the presence of catecholamines both in vivo and in vitro. There was a small increase in the number of beta-adrenergic receptors but no change in the amount of the two isoforms of the stimulatory GTP-binding protein, Gs. Treatment with somatotropin decreased the response to antilipolytic agents such as the adenosine analog N6-phenylisopropyladenosine and prostaglandin E1. There was, however, no change in the number of adenosine receptors or amounts of the inhibitory GTP-binding proteins (Gi-1 plus Gi-2). Somatotropin also decreased prostaglandin E2 production by subcutaneous adipose tissue in vivo. Somatotropin treatment thus alters lipolytic regulation in sheep and this is characterized by changes in a number of proteins involved in this process. PMID- 8707712 TI - Effects of animal age, marbling score, calpastatin activity, subprimal cut, calcium injection, and degree of doneness on the palatability of steaks from limousin steers. AB - Strip loin (longissimus lumborum), sirloin (gluteus medius) and round (semimembranosus) subprimals from 114 purebred and crossbred Limousin steers were used to identify main effects and interactions of animal age, marbling score, calpastatin activity, subprimal cut, calcium injection (5% wt/wt with a 200 mM CaCl2 solution at 48 h postmortem), and degree of doneness on the palatability of cooked beef steaks. Steaks were aged for 14 d, frozen, thawed, cooked to different internal temperature end points, visually scored for degree of doneness, sheared on a Warner-Bratzler shear machine, and evaluated by a trained taste panel. Raw and cooked steaks from carcasses of higher USDA quality grades had higher fat and lower moisture percentages (P < .05). Higher degrees of doneness resulted in lower moisture percentages (P < .05). Lower shear force values were associated with less variation in shear force. Younger slaughter age and lower calpastatin activity both resulted in greater tenderness (P < .05). Shear force was lowest between "medium rare" and "medium" and increased toward both ends of the degree of doneness scale for round and sirloin steaks; however, shear force increased linearly with degree of doneness in strip loin steaks (P < .05). Subprimal cut had the largest effect on taste panel tenderness ratings, and degree of doneness had the largest effect on taste panel juiciness ratings. The improvement in shear force due to CaCl2 injection was greater for strip loin and sirloin steaks than for round steaks (P < .05 for the interaction). Injection with CaCl2 improved all taste panel attributes. In addition, CaCl2 injection reduced the toughening effects of cooking (P < .05). PMID- 8707713 TI - Crystalline lysine and threonine supplementation of soft red winter wheat or triticale, low-protein diets for growing-finishing swine. AB - Five trials, with five treatments each, involving a total of 240 pigs were conducted to evaluate the effectiveness of supplementation of soft red winter wheat- or triticale-based diets with crystalline lysine and threonine for growing and finishing pigs (25 to 110 kg). Within each trial, wheat- or triticale-based diets were supplemented with soybean meal to meet the requirement of the first- (lysine) or third- (isoleucine/methionine + cystine) limiting amino acid. Diets formulated to the third-limiting amino acid also were supplemented with feed grade L-lysine. HCl and L-threonine to meet the requirements of the first- and second-limiting amino acids. A cornsoybean meal diet was included in each trial as a positive control treatment resulting in the five dietary treatments. Three different crop years of soft red winter wheat and triticale were utilized. Three of the trials, conducted in successive years with each using a different crop year of grain, were done at Marianna, and the other two, conducted in successive years using yr 1 and 2 crops, respectively, were done at Gainesville. In all trials, growing diets (.82% lysine) were fed from 29 (Marianna) or 25 kg (Gainesville) to 55 kg average BW and finishing diets (.64% lysine) to 110 or 100 kg. Over the three crop years, the wheat and triticale averaged 11.3 and 11.0% crude protein, .36 and .38% lysine, and .36 and .37% threonine, respectively. Overall, at either location, growth rate and carcass lean content were not affected by grain source (P > .10). At Marianna, gain:feed was 2 to 3% lower for pigs fed the triticale diets than for pigs fed wheat (P = .15) or corn (P < .10); gain:feed was not affected by grain source at Gainesville (P > .10). At both locations, substantial replacement of soybean meal protein with crystalline lysine and threonine did not affect pig growth or carcass lean content (P > .10), even when the diet contained very little or no soybean meal as occurred for finishing pigs (55 to 110 kg) fed diets containing triticale. PMID- 8707714 TI - In vivo glucocorticoid effects on porcine natural killer cell activity and circulating leukocytes. AB - Porcine natural killer (NK) cell cytotoxicity, plasma cortisol, total white blood cells (WBC), neutrophil:lymphocyte ratio (N:L), and circulating blood leukocytes were examined from pigs injected i.v. with either saline, ACTH, cortisol, or treated with metyrapone. Plasma cortisol increased (P < .05) after ACTH and cortisol treatments and decreased (P < .05) after metyrapone treatment; thus, treatments had the intended effects on in vivo cortisol concentrations. In Exp. 1, pigs were injected with either saline or ACTH at 0600 after the initial blood samples were taken (time 0). The ACTH had no effect (P > .10) on NK cytotoxicity. Pigs injected i.v. with ACTH had fewer lymphocytes and more neutrophils (P < .05) than control pigs. The N:L ratio was greater (P < .05) among ACTH-injected than among control pigs. In Exp. 2, pigs were injected i.v. with either saline or 40 or 400 micrograms of cortisol at 0600 after the initial blood samples were obtained (time 0). Cortisol at 40 micrograms had no effect (P > .10) on NK cytotoxicity. However, a 400-micrograms bolus of cortisol reduced (P < .05) NK cytotoxicity (control = 39.5, cortisol = 28.3% cytotoxicity, SEM = 3.7). Each dose of cortisol reduced (P < .05) circulating blood lymphocyte numbers. In Exp. 3, pigs were fed 1 g of metyrapone or no metyrapone the night before sampling. Blood samples were obtained at 0600, 0700, and 0800. Metyrapone reduced (P < .05) NK cytotoxicity (control = 28.6, metyrapone = 11.8%, SEM = 1.9). Pigs treated with metyrapone had greater (P < .05) numbers of neutrophils than control pigs. Numbers of lymphocytes were greater (P < .05) among control than among treated pigs. Pigs treated with metyrapone had a greater (P < .05) N:L ratio than control pigs. In conclusion, normal physiological concentrations or moderately increased blood cortisol concentrations did not influence NK activity, although leukocyte distributions were changed. We conclude that greatly increased or greatly decreased circulating concentrations of cortisol reduced NK cytotoxicity. PMID- 8707715 TI - Calving intervals in beef cows at 2, 3, and 4 years of age when breeding is not restricted after calving. AB - The objective of this study was to evaluate calving intervals and repeatability of calving intervals of young beef cows when breeding seasons were initiated immediately after calving. Average calving date at 2, 3, and 4 yr of age was recorded, and duration of calving interval between the first and second and second and third parities was compared in 178 crossbred beef cows calving between 1981 and 1988. Cows were placed in paddocks with intact bulls immediately after calving so they had the opportunity to conceive at the estrus associated with the initial estrous cycles after calving. Average Gregorian date of calving for cows was progressively earlier with older age (March 9, March 5, and February 14 at 2, 3, and 4) yr of age, respectively). Repeatability of calving intervals within individual cows was also evaluated. There was an interaction between year and parity on calving interval between 2 and 3 (P < .001) and 3 and 4 (P < .004) yr of age. Repeatability of calving interval was low and negative during 5 of the 6 yr. In young beef cows, year and parity interact to influence calving interval, and this average interval is less than 365 d when initiation of the breeding season is not restricted after calving. PMID- 8707716 TI - N-methyl-D,L-aspartate-induced growth hormone secretion in barrows: possible mechanisms of action. AB - Four experiments were conducted to determine mechanisms by which n-methyl-d,l aspartate (NMA) increases serum concentrations of growth hormone (GH). Blood samples were collected from barrows every 15 min for 2 h (Exp. 1, 2, and 3) or 3 h (Exp. 4) immediately before and immediately after i.v. treatments. In Exp. 1, barrows (n = 4/treatment) received either .9% saline or 1.25, 2.5, or 5 mg of NMA/kg of BW. The change in circulating GH concentrations was greater (P < .05) for barrows receiving 2.5 mg (by 883%) or 5.0 mg of NMA/kg of BW (by 1,095%) than for those injected with saline. In Exp. 2, barrows (n = 4/treatment) received NMA (2.5 mg/kg of BW) or injections of 1.25 mg of the pure d or pure 1 isomers of NMA/kg of BW. Growth hormone concentrations increased by 177% (P < .025) after NMA treatment and by 245% (P < .01) after injection of the pure d isomer of NMA. The pure 1 isomer of NMA had no effect (P > .1) on GH concentrations. In Exp. 3, barrows received NMA (2.5 mg/kg of BW) 10 min after i.m. injection of saline (n = 7) or ketamine hydrochloride ( n = 8; 19.9 mg/kg of BW), an n-methyl-d-aspartate (NMDA) receptor antagonist. The NMA increased (P < .01) GH concentrations by 289% in saline-pretreated barrows but had no effect (P > .1) on barrows pretreated with ketamine hydrochloride. In Exp. 4, barrows (n = 4/treatment) received NMA 3 h after i.v. pretreatment with antisera to GH-releasing factor (GRF; 154 mL) or no pretreatment. Serum GH concentrations increased by 166% (P < .05) after injection of NMA in barrows receiving no pretreatment. The NMA had no effect (P > .1) on GH concentrations in barrows receiving antisera to GRF. Our results support the concept that NMDA stimulates GRF, and hence GH secretion, by activating an NMDA receptor. PMID- 8707717 TI - Effect of prepubertal retinyl palmitate treatment on uterine development and subsequent uterine capacity in swine. AB - The effect of prepubertal retinyl palmitate treatment on uterine development and uterine capacity was determined. Gilts remained intact or were unilaterally hysterectomized-ovariectomized (UHO) on d 28, 84, or 112 of age. Starting at birth, half of the gilts in each UHO group were treated with retinyl palmitate (14,000 IU.kg-1.wk-1) and half received control carrier solution. All intact gilts received control carrier solution. Blood samples (d 28, 56, 84, and 112 of age) were analyzed for retinoid and retinol-binding protein (RPB). At UHO, part of the excised uterine horn was processed for morphometric analysis of the uterine wall, and tissues were incubated in minimal essential medium with [3H]leucine to assess total protein and RBP production. After puberty, gilts were mated and then killed on d 44 to 47 of pregnancy; uterine length, number of corpora lutea and fetuses, and fetal, placental, and empty uterine weights were recorded. Endometrial tissues were incubated in minimal essential medium with [3H]leucine, and nondialyzable radioactivity, acid phosphatase, and RBP production were measured. During the prepubertal period, retinyl palmitate treatment decreased (P < .05) myometrial area (longitudinal and circular), but it had no effect on other components of the uterine wall or production of nondialyzable radioactivity or secretion of RBP by uterine tissue. During pregnancy, prepubertal retinyl palmitate treatment increased (P < .05) uterine length but had no effect on uterine weight, number of fetuses, placental or fetal weights, or endometrial production of nondialyzable radioactivity, acid phosphatase, or RBP. Therefore, prepubertal retinyl palmitate treatment at the dosage administered had no effect on uterine capacity. PMID- 8707718 TI - Embryonal survival to 6 days in mice selected on different criteria for litter size. AB - Embryonal survival was compared in mice resulting from four criteria of selection: LS = selection on number born; IX = selection on an index of ovulation rate and ova success; UT = selection on number born to unilaterally ovariectomized females; and LC = unselected control. Selection occurred for 21 generations with three replicates of the four criteria; thereafter, relaxed selection was practiced. The evaluation was performed using mice of two replicates at Generation 35 and one replicate at Generation 36. Data on a total of 289 female mice were recorded. Females, at an average age of 9 wk, were mated to males of the same line. Six days after mating, each female was killed, ovaries were excised, corpora lutea were counted and equated to number of ova shed, and the numbers of implantation sites in each uterine horn were recorded. Least squares means were .84, .91, .85, and .82 for left embryonal survival (left implantations/left ova) and .91, .90, .86, and .87 for right embryonal survival for LS, IX, UT, and LC, respectively. The right side had greater ovulation rate (P < .001) and number of implantations (P < .001). For embryonal survival, the criterion x side interaction was possibly important (P < .09). Selection for litter size by different criteria increased ovulation rate (P < .003) and embryonal survival (P < .05) to 6 d. However, responses in embryonal survival were not greater after UT selection compared with LS or IX selection. PMID- 8707719 TI - Relationship between circulating progesterone and conception at the first postpartum estrus in young primiparous beef cows. AB - The objective of this study was to determine whether patterns of change in concentration of progesterone in circulation of primiparous beef cows were related to conception rates at the first postpartum estrus. Data were collected over 2 yr from crossbred primiparous 2-yr-old cows (n = 77). Cows were artificially inseminated when estrus was detected between 3 and 20 (yr 1) or 3 and 23 (yr 2) wk after parturition. Cows were exposed to intact bulls for 32 (yr 1) or 21 d (yr 2) after the periods of AI, and estrus detection also occurred during the period when natural mating occurred. After parturition, blood samples were collected twice weekly to determine time of transient (less than 1 wk) and sustained increases (more than 1 wk) in progesterone of more than 1 ng/mL of serum. Mean times from parturition to transient increase in progesterone, first estrus, conception, and sustained increase in progesterone were 91, 100, 108, and 105 d, respectively. Sixty-four percent of the cows conceived as a result of AI at the first estrus that was detected after parturition, 32% conceived at a subsequent estrus, and 4% failed to conceive. In 31.1% of the cows, a transient increase in progesterone was not detected before the first estrus after calving. Conception as a result of AI at the first estrus after calving was less if a transient increase in progesterone did not precede estrus (76 vs 41%; P < .01). These data indicate that the majority of cows have increases in progesterone before the first postpartum estrus. If the transient pre-estrus increase in progesterone occurs in postpartum cows, there seems to be an enhanced conception rate as compared with cows without the increase in progesterone before their first estrus following parturition. PMID- 8707720 TI - Effect of nutrition on embryonal mortality in gilts: association with progesterone. AB - The effect of the timing of nutritional changes during the immediate period after mating on early embryonal survival and of progesterone as a potential mediator of such changes was studied. A total of 82 gilts were initially fed 2.5 kg.gilt-1.d 1 for one estrous cycle before they were inseminated at 16 and 24 h after the onset of estrus (d 0) using fresh, pooled semen. After AI, gilts were randomly allocated to one of the three feeding regimens, normal NRC allowance of 1.5 x maintenance per day from d 1 (Group N1) or d 3 (Group N3) or an allowance of 2 x maintenance from d 1 (Group H1). All gilts were fed on an individual basis. Single blood samples were collected 72 h after first detection of standing estrus. From d 15 onward, all gilts were fed 1.8 kg/d until they were slaughtered on d 28 +/- 3. Total and viable empryonal survival were affected by dietary treatment (P = .044 and .027, respectively), and viable embryonal survival in group N1 was greater than in group H1 (84.7 +/- 4.5 vs 64.5 +/- 7.6%; P < .05). Plasma progesterone was greater in group N1 than in groups N3 and H1 (10.5 +/- 1.0 vs 3.7 +/- .8 and 4.5 +/- .7 ng/mL, respectively; P < .05). The timing of the change in feed allowance after mating is therefore crucial for demonstrating effects of nutrition on embryonal survival in gilts, and progesterone may mediate these effects. PMID- 8707721 TI - Forage systems for beef production from conception to slaughter: III. Finishing systems. AB - Fall-weaned Angus calves grazed or were fed different forages during winter followed by 1) N-fertilized tall fescue (Festuca arundinacea Schreb.) grazed alone, 2) bluegrass (Poa pratensis L.)-white clover (Trifolium repens L.) sequence grazed with tall fescue-red clover (Trifolium pratense L.), or 3) bluegrass-white clover sequence grazed with alfalfa (Medicago sativa L.) orchardgrass (Dactylis glomerata L.). Heifers were supplemented with grain at 1% of BW from April until slaughter in July. One-half of steers were supplemented with grain at 1% of BW from July until slaughter in October. Remaining steers were fed no grain but were finished on corn silage supplemented with .9 kg of soybean meal per steer daily, from October until slaughter in late January. Including alfalfa-orchardgrass in systems during the finishing phase resulted in higher daily and total gains during the grazing period, and carcasses had more marbling and higher USDA quality grades at slaughter compared with carcasses of cattle on systems using fescue-red clover. Correlation of final weight with carcass characteristics was low (r < .5). Performance and carcass characteristics were influenced as much or more by forage consumed during the previous wintering phase as by forage fed during the finishing phase. Wintering cattle on stockpiled fescue-alfalfa or alfalfa-orchardgrass hay generally resulted in higher BW at slaughter and more desirable carcass characteristics than systems using tall fescue alone or in combination with red clover. This was particularly notable in steers that grazed without grain until October and were finished on corn silage plus supplement. Final BW and carcass characteristics in all cattle were improved by full season grazing followed by feeding corn silage, compared with cattle finished with grain on pasture. PMID- 8707722 TI - Evaluation of the potential of supplements to substitute for low-quality, tallgrass-prairie forage. AB - Thirteen ruminally fistulated steers (260 +/- 15 kg) were used in an incomplete Latin square with 13 treatments and four periods to evaluate the potential for different supplements to substitute for intake of low-quality, tallgrass-prairie forage. Steers were given ad libitum access to forage and received either no supplement (control = CTL) or one of four supplements, each fed at three different levels of intake. Supplements included 1) moderate CP (17.5%) concentrate (MCP-CON), 2) high CP (32.7%) concentrate (HCP-CON), 3) long-stem alfalfa hay (LSAH; 17.2% CP), and 4) alfalfa pellets (AP; 16.3% CP). Concentrates were mixtures of sorghum grain and soybean meal. Supplements were fed to supply .05, .10, and .15% BW of CP/d. Forage intake and digestible DMI were increased (P < .01) for supplemented steers compared with CTL (22 and 96%, respectively). Steers receiving increasing MCP-CON exhibited a quadratic (P = .03) forage intake response. Offering MCP-CON higher than .10% BW of CP/d (approximately .59% BW of DM) resulted in a substitution rate of -.56 g of forage/g of supplement. Although effects were not significant for steers receiving LSAH, the decline in forage intake at the high level of supplementation (-.48 g of forage/g of supplement) was similar in magnitude to that for MCP-CON. Forage intake increased linearly (P < .01) across supplementation levels for HCP-CON and AP supplements. Passage rates were faster (P < .01) for supplemented steers. However, passage rates for HCP-CON and AP groups increased linearly (P < .01) with increasing supplement, whereas they declined at the highest supplementation rate for MCP-CON and LSAH (quadratic, P < or = .05). Most fermentation variables displayed positive responses to supplementation per se and to increasing amount of supplements offered. In conclusion, although supplementation effectively enhances the use of low-quality forage, supplement type may affect the likelihood of observing substitution effects. PMID- 8707723 TI - Elucidation of factors associated with the maturity-related decline in degradability of big bluestem cell wall. AB - Big bluestem (Andropogon gerardii) forage samples were collected from three ungrazed, annually burned pastures at 38, 58, and 97 d after burning. Cell wall material received five treatments: chlorite delignification, chlorite delignification plus alkali extraction, NaOH, NaOCH3 in methanol, or NaBH4. Untreated and treated cell walls were analyzed for carbohydrate composition (glucose, xylose, arabinose, galactose, and uronic acids), acetyl bromide lignin, acid detergent lignin, alkali-labile phenolics (p-coumaric and ferulic acids), acetyl groups, and 24- and 72-h in vitro degradabilities of neutral monosaccharides. A number of compositional features, notably concentrations of xylose, core lignin as measured by acetyl bromide lignin, alkali-labile phenolics, and acetyl groups, were well related to the decline in cell wall degradability that occurred with increasing maturity of big bluestem. p-Coumaric acid increased with increasing maturity to a greater extent than did ferulic acid. Acid detergent lignin was not well related to degradability of the cell wall for either the untreated or chemically treated cell walls. Chemical treatments failed to identify any particular cell wall component as being most inhibitory. However, all treatments improved in vitro degradability of the carbohydrate fraction, indicating that components contributing to the undegradability of big bluestem cell wall are sensitive to chemical alteration. Treatments involving alkali were most effective for improving degradability of big bluestem cell walls. PMID- 8707724 TI - Technical note: a simple, rapid assay for alpha-amylase in bovine pancreatic juice. AB - A procedure is described for the determination of alpha-amylase activity in bovine pancreatic juice. Bovine pancreatic juice was incubated with potato amylopectin for 6 min, and then alkaline potassium ferricyanide was added to stop the enzymatic reaction. Tubes were placed in a boiling water bath for 10 min and then cooled, and 40 mL of distilled water was added. Optical density was read at 420 nm, and the amount of end product (reducing sugars) was determined by comparison with maltose standards. Ten dilutions of bovine pancreatic secretion were assayed to determine the intraassay precision of this method, resulting in a CV of 1.05%. A pooled sample was assayed five times over five separate days to determine the interassay precision, resulting in a CV of 7.3%. This alpha-amylase assay provides a repeatable, rapid, and economical determination of alpha-amylase activity from bovine pancreatic juice. PMID- 8707725 TI - Spent cellulose casings as potential feed ingredients for ruminants. AB - Cellulose casings are used to contain and form meat and poultry emulsions during the smoking and cooking process. Casings then are stripped from the cooked product and traditionally disposed of in landfills. Because of the bulk of the spent cellulose casings (SCC), rapid composting technology may be used to reduce bulkiness. The following SCC were evaluated in vitro and in vivo: fibrous ground (FG), fibrous composted (FC), NOJAX ground (NG), and NOJAX composted (NC). In vitro digestibility was determined by incubating SCC with mixed ruminal bacteria for 0, 6, 12, 24, 36, 48, and 72 h. In vivo data were collected using four ruminally cannulated Holstein steers in a 4 x 4 Latin square design. Diets consisted of a 50:50 ratio of alfalfa hay-wheat middlings with 5% cornsteep liquor. Diets contained no SCC (CON) or 25% (DM basis) of the FC, FG, or NC SCC substrate. Casings were high in structural carbohydrate and salt content but low in CP, ether extract, and lignin concentrations. In vitro OM digestibility at 24 h was highest (P < .05) for FC and lowest (P < .05) for NG; FG and NC were intermediate. Composting tended to reduce fiber content and increase digestion. In vivo intakes and digestibilities were not adversely affected by inclusion of SCC in the diet. Thus, SCC have the ability to partially replace more traditional forages, such as alfalfa hay and wheat middlings, in high-fiber diets for growing beef cattle. Limitations in the use of SCC as a partial replacement of traditional feedstuffs will likely be because of high salt concentrations in the casings resulting from product brine chilling. PMID- 8707726 TI - Vitamin E supplementation and stress affect tissue alpha-tocopherol content of beef heifers. AB - The effect of stress on tissue alpha-tocopherol was investigated in 16 crossbred heifers fed a corn/corn silage-based diet. For 28 d, eight heifers (379 +/- 10 kg BW) received a dietary supplement of 1,000 IU of dl-alpha-tocopheryl acetate, whereas the controls (375 +/- 10 kg BW) received no supplemental vitamin E. Tissue samples of plasma, red blood cells, liver, trapezius, and longissimus muscles and subcutaneous fat immediately dorsal to each muscle were taken on d 1 for determination of alpha-tocopherol concentration. On d 2 through 4 each heifer was restricted to 2.61 kg of grass hay and allowed water. On d 5, 6, and 7 no feed or water was given, 100 IU of ACTH and .0024 mg of epinephrine/kg BW were given every 8 h, and biopsies for alpha-tocopherol content were again taken on d 7. The stress reduced (P < .01) mean BW, increased (P < .01) serum cortisol, creatine kinase, and urea. After stress, supplemental vitamin E reduced (P < .13) the increase in creatine kinase relative to that in heifers not supplemented with vitamin E. Stress also increased (P < .04) serum Se in heifers fortified with the vitamin E. Alpha-tocopherol content of plasma, red blood cells, liver, and subcutaneous fat dorsal to the trapezius muscle was increased (P < .01) by supplemental vitamin E. The stress treatment reduced (P < .01) alpha-tocopherol content of plasma in those fed the vitamin E and increased it (P < .05) in the nonsupplemented vitamin E-deficient heifers. Stress also decreased red blood cell (P < .01) and liver (P < .05) alpha-tocopherol content in cattle supplemented with vitamin E. Tissue alpha-tocopherol concentrations were reduced by stress only when a diet adequate in vitamin E was fed. In addition, in most sampled tissues, stress did not affect alpha-tocopherol concentrations. PMID- 8707727 TI - Interactions between Fibrobacter succinogenes, Prevotella ruminicola, and Ruminococcus flavefaciens in the digestion of cellulose from forages. AB - The synergistic and inhibitory interactions observed between Fibrobacter succinogenes A3c, Prevotella ruminicola H2b, and Ruminococcus flavefaciens B34b in the digestion of forage cellulose were studied in detail. Orchardgrass and alfalfa hays, both at two maturity stages, were used as substrates. Sequential inoculation procedures were developed whereby a second inoculation was made after the initial fermentation was killed. Total cellulose digestion from sequential addition of the organisms was then compared to values obtained in simultaneous co culture. When the noncellulolytic P. ruminicola was co-cultured with either of the two cellulolytic species (F. succinogenes or R. flavefaciens) forage cellulose digestion numerically increased over that of the cellulolytic species alone. In contrast, decreases from co-culture values were noted with sequential addition of the organisms. When F. succinogenes and R. flavefaciens were co cultured, cellulose digestion was reduced compared to F. succinogenes alone. However, no such reduction was observed when the organisms were added sequentially. Further experiments indicated that this inhibitory activity is only produced when the organisms are co-cultured and is stable to autoclaving at 121 degrees C for 20 min. Inhibition of this type could be the result of bacterocin production by one of the organisms; however, most bacterocins are destroyed by autoclaving. PMID- 8707728 TI - Characterization of the proteolytic activity of commercial proteases and strained ruminal fluid. AB - The objective of this research was to formulate a mixture of commercial proteases that would mimic the rate and extent of protein degradation obtained using strained ruminal fluid. The proteolytic activity of strained ruminal fluid and several commercial proteases was characterized using 13 L-amino acid p nitroanilides as artificial substrates. A mixture of Streptomyces griseus protease, chymotrypsin, and proteinase K at .042, 2.5, and .5 enzyme units/mL, respectively, was similar to the activity of strained ruminal fluid against the same artificial substrates. However, degradative activities were different in incubations with feed proteins as substrates. The rates of degradation of expeller soybean meal, solvent soybean meal, and casein were .08, .05, and .08/h, respectively, using the enzyme mixture and .03, .15, and .24/h using strained ruminal fluid. A second experiment compared degradative activity of S. griseus protease at .066 enzyme units/mL, ficin at .5 enzyme units/mL, and a mixture of trypsin, carboxypeptidase B, chymotrypsin, and carboxypeptidase A at 116.6, .5, 2.5, and .5 enzyme units/mL, respectively. Protein degradation rates obtained with strained ruminal fluid were two to six times faster than those obtained with the enzyme mixtures. A third experiment compared the degradability of 15 feed proteins with the mixture of trypsin, carboxypeptidase B, chymotrypsin and carboxypeptidase A to that with strained ruminal fluid. Degradation rates obtained using strained ruminal fluid ranged from .007 to .217/h; degradation rates using the enzyme mixture ranged from .010 to .079/h and were lower (P = .004) than with strained ruminal fluid. Overall, the experiments indicated that the commercial enzymes tested did not mimic the protein degradative activity of strained ruminal fluid. PMID- 8707729 TI - Effects of varying levels of duodenal or midjejunal glucose and 2-deoxyglucose infusion on small intestinal disappearance and net portal glucose flux in steers. AB - Six crossbred steers (261 +/- 18 kg BW) fitted with hepatic portal, mesenteric venous and arterial catheters, and duodenal, midjejunal, and ileal cannulas were used in a replicated 3 x 3 Latin square design to determine the effect of varying levels and site of glucose plus 2-deoxyglucose (2DG) infusion on net portal drained visceral flux. Steers were fed chopped alfalfa in six equal portions daily at 1.5% of BW. Glucose (0, 9, or 18 g/h) and 2DG (0, 1, or 2 g/h) were infused continuously through the duodenal or midjejunal cannula (two infusion sites) at total glucose plus 2DG infusion rates of 0, 10, or 20 g/h. Arterial and portal blood samples were taken simultaneously at 20-min intervals from 5 to 9 h of infusion. Portal blood flow was determined by continuous infusion of p aminohippurate and net flux was calculated as venous-arterial concentration (PA) difference times blood flow. Arterial concentration of glucose was not affected (P > .10) by glucose plus 2DG infusion, whereas arterial concentration of 2DG was greater (P < .05) when glucose plus 2DG was infused into the duodenum and increased (linear, P < .10) as amount of glucose plus 2DG infused into both the duodenum and midjejunum increased. Net portal flux and PA difference of glucose and 2DG were greater (P < .05) when glucose plus 2DG was infused into the duodenum. Although 2DG was infused at 10% of the total glucose plus 2DG infusion, it accounted for only 1.7 and .7% of the glucose plus 2DG appearing in portal blood when glucose plus 2DG was infused at 10 and 20 g/h, respectively. We conclude that glucose is more readily absorbed across the proximal-half than the distal-half of the small intestine, and that passive diffusion is a minor route of glucose absorption. PMID- 8707730 TI - Clinical studies in chronic bronchitis: a need for better definition and classification of severity. PMID- 8707731 TI - The comparative efficacy and safety of teicoplanin and vancomycin. AB - Glycopeptide antibiotics, such as teicoplanin and vancomycin, are active against staphylococci (including methicillin resistant strains), streptococci, enterococci and Clostridium spp. Vancomycin and teicoplanin are both widely used in the treatment of infections caused by Gram-positive organisms. Vancomycin can, however, provoke a number of side-effects, and serum concentrations should be monitored during treatment. Teicoplanin has a longer half-life than vancomycin, it can be given as an intravenous bolus or by intramuscular injection, and nephrotoxicity and ototoxicity are relatively uncommon. Treatment with teicoplanin might, therefore, offer advantages over treatment with vancomycin provided that similar clinical efficacy can be shown. At least 11 clinical trials comparing the efficacy and safety of teicoplanin and vancomycin have been carried out worldwide. Meta-analysis of the combined results from these studies indicates that more than three-quarters of the patients in each of the treatment groups had a clinical response to therapy. Meta-analysis of the numbers of adverse events occurring in each treatment group shows significantly fewer reports of adverse events in patients receiving teicoplanin (13.9%) than in those receiving vancomycin (21.9%). Direct comparisons are difficult because of inherent differences between studies, but available data suggest that teicoplanin is as effective as vancomycin and that its superior tolerability together with advantages such as once-daily bolus administration, intramuscular use and lack of requirement for routine serum monitoring, give it considerable potential for use in clinical practice. PMID- 8707732 TI - A Tn1545-like transposon carries the tet(M) gene in tetracycline resistant strains of Bacteroides ureolyticus as well as Ureaplasma urealyticum but not Neisseria gonorrhoeae. AB - The presence of the tet(M) determinant and conjugative transposons related to Tn1545 in urogenital pathogens, Bacteroides ureolyticus (10 strains), Neisseria gonorrhoeae (37 strains) and Ureaplasma urealyticum (81 strains), was studied by PCR analysis and hybridization assay. All tetracycline-resistant strains that hybridized with a probe specific for tet(M) gave the expected fragment when their DNA's were subject to PCR. The tet(M) gene and int-Tn, the gene encoding the protein required for the movement of Tn1545-like transposons, were always found together in tetracycline resistant strains of B. ureolyticus and U. urealyticum. In contrast, int-Tn was not detected in tet(M)-mediated tetracycline-resistant N. gonorrhoeae strains. These data suggest carriage of tet(M) by a Tn1545-like transposon in some tetracycline-resistant B. ureolyticus. PMID- 8707733 TI - Quantitative comparison in vitro of mutational antibiotic resistance of Enterobacter spp. using a spiral plater. AB - The presence of spontaneous mutational antibiotic resistance among 18 bacteremic isolates of Enterobacter spp. to cefotaxime, ceftazidime, gentamicin, amikacin, ciprofloxacin, and trimethoprim-sulfamethoxazole was determined quantitatively in vitro using a spiral plater. Each drug was delivered using the device and the agar plates were inoculated in radial streaks. The degree of resistance was estimated by dividing the antimicrobial concentration required to inhibit 90% of the colonies growing in the area beyond the MIC by the MIC itself. The degree of resistance to third-generation cephalosporins and aztreonam was statistically significantly greater than that to co-trimoxazole, imipenem, and ciprofloxacin (P < 0.01); the latter three antibiotics showed virtually no mutational resistance. An intermediate level of resistance was induced by aminoglycosides, and mutational resistance to piperacillin varied between this and the higher levels observed for the cephalosporins. By providing a simple and efficient means of detecting spontaneous mutational resistance, the spiral plater may prove useful in identifying those antimicrobial agents which induce few or no mutants and therefore may be more likely to be successful in treating infections due to Enterobacter spp. PMID- 8707734 TI - In-vitro susceptibility of isolates of methicillin-resistant Staphylococcus aureus 1988-1993. AB - MICs for 423 strains of methicillin-resistant Staphylococcus aureus (MRSA) isolated in Hong Kong during 1988-1993 were performed for 15 antimicrobial agents: erythromycin, chloramphenicol, tetracycline, minocycline, gentamicin, netilmicin, trimethoprim, rifampicin, fusidic acid, ciprofloxacin, vancomycin, teicoplanin, sparfloxacin, clinafloxacin and RP 59500 (quinupristin/dalfopristin). Susceptibility to antibiotics generally remained stable throughout the study period, with the exception of the quinolones. Resistance to ciprofloxacin (breakpoint 4 mg/L) increased from a low of 9% in 1988 to a high of 82% in 1993. For sparfloxacin the corresponding figures were 9% and 78%, respectively. Six (1%) clinafloxacin-resistant strains were found. MIC50s and MIC90s of clinafloxacin increased from < or = 0.06 mg/L and 0.25 mg/L in 1988 to 1.0 mg/L and 2.0 mg/L, respectively, in 1993. All 423 strains were phage typed (typability 70%) and a diversity of phage types which changed during the observation period, with 13 dominating types, was observed. Ciprofloxacin resistance occurred in 12 of the dominating types, in 46 non-typable strains, and also in 23 strains of different, sporadically occurring types, indicating that the emergence of quinolone resistance was not due to dissemination of a single or few MRSA clones. The usefulness of quinolones in the treatment of MRSA infections is likely to be seriously constrained by the emergence of resistance. MICs for RP 59500 were < or = 2 mg/L for all isolates, suggesting that this agent merits further evaluation as an anti-MRSA agent. All MRSA remained susceptible to vancomycin and teicoplanin throughout the study period. PMID- 8707735 TI - The effect of antibiotics on bacteria under hyperbaric conditions. AB - The sensitivity of selected bacteria to a range of antibiotics was tested under hyperbaric conditions used in saturation diving. The effect of hyperbaric helium and oxygen (heliox) on antibiotic stability and on induction of beta-lactamase was also determined. Increased resistance to penicillin (up to 23%) was shown by Staphylococcus aureus and to gentamicin (up to 46%) and rifampicin (up to 18%) by Escherichia coli and Salmonella typhimurium at 36 and 71 bar pressure. Exposure to 71 bar heliox did not affect antibiotic activity but increased the production of beta-lactamase in inducible S. aureus and Bacillus subtilis and production of beta-galactosidase in inducible E. coli. Increased resistance to antibiotics in saturation diving conditions can be attributed in some cases to the influence of hyperbaric pressure on induction mechanisms in bacteria. The experimental system devised for this work is suitable for more detailed examination of the influence of hyperbaric stress on antibiotic resistance and of its effect on induction mechanisms in general. PMID- 8707736 TI - Antifungal susceptibility testing using the E test: comparison with the broth macrodilution technique. AB - The National Committee for Clinical Laboratory Standards reference broth macrodilution method for antifungal susceptibility testing was compared with the E test by testing 86 clinical isolates of Candida spp. and Cryptococcus neoformans. The MIC agreement rates for the two methods for Candida spp. were 73 89% within +/-1 doubling dilution and 87-100% within +/-2 dilutions. For C. neoformans, agreement within +/-1 dilution was > 70% for all the agents tested except fluconazole for which agreement was 35%. Our data support the further evaluation of the E test as an alternative method for antifungal susceptibility testing. However, E test MICs of fluconazole for C. neoformans, particularly C. neoformans var. gattii should be interpreted with caution, as falsely elevated MICs may occur. PMID- 8707737 TI - Relative growth measurement of Candida species in a single concentration of fluconazole predicts the clinical response to fluconazole in HIV infected patients with oral candidosis. AB - The growth of 113 Candida spp. isolated prospectively from 104 HIV-positive patients treated for thrush was determined using a single concentration of 3 mg/L (10(-5) M) fluconazole relative to growth in drug free medium. Using a receiver operator characteristic curve, a relative growth in fluconazole of > or = 88% best discriminated between isolates from 56 patients who responded to treatment with fluconazole and 48 who failed to respond to 7 days, treatment with at least 100 mg/d drug with a sensitivity of 98% and a specificity of 96%. When the isolates from the eight patients with mixed infections due to both resistant and susceptible yeasts were excluded, the sensitivity and specificity both reached 100%. Relative growth of Candida spp. in a single concentration of fluconazole can therefore be used to accurately predict the clinical response to standard fluconazole treatment of thrush in patients infected with HIV. PMID- 8707738 TI - Comparative activity of piperacillin/tazobactam against 5625 isolates from hospitalised patients. Multicentre Study Group. AB - The comparative activities of piperacillin/tazobactam and other antibiotics against 5625 Gram-negative and Gram-positive bacteria freshly isolated from hospitalized patients was examined. The percentage of Enterobacteriaceae susceptible to piperacillin/tazobactam, ceftazidime, imipenem and ciprofloxacin was very similar and varied from 81% to 84% in the Enterobacteriaceae with inducible type 1 beta-lactamases and from 93 to 97% in the non-inducible organisms. Piperacillin/tazobactam was similar in activity to ceftazidime against Pseudomonas aeruginosa and more active than imipenem or ciprofloxacin. Piperacillin/tazobactam is more active than ceftazidime against Gram-positive bacteria (staphylococci, enterococci) and seems to be a promising antibiotic for nosocomial infections. PMID- 8707739 TI - An 8 year Microbe Base survey of the epidemiology, frequency and antibiotic susceptibility of Pseudomonas aeruginosa hospital isolates in the United Kingdom. AB - Over the last 8 years selected United Kingdom hospitals have contributed antimicrobial susceptibility data to a central data base called Microbe Base. During that period, data on 1,000,067 isolates have been collected, including 29,425 isolates of Pseudomonas aeruginosa. The present study focused on the epidemiology, frequency and antibiotic susceptibility of P. aeruginosa isolates (17,440) from hospitalised patients. In such patients P. aeruginosa was predominant in general surgery (20%), care of the elderly (18%), general medicine (13%), and paediatrics and neonates (10%). Ninety-five per cent of P. aeruginosa were susceptible in vitro to ceftazidime compared with 93% for piperacillin, 92% for gentamicin. 90% for ciprofloxacin, and 89% for imipenem. Excluding susceptibility data reported in 1991, there was no change in the susceptibility pattern of P. aeruginosa throughout the study period. PMID- 8707740 TI - Activity of penciclovir in antiviral assays against herpes simplex virus. AB - The effect of penciclovir and acyclovir on the replication of herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) strains was determined in MRC-5 cells infected with 0.01 pfu/cell and exposed to the drugs for 72 h to allow multiple cycles of replication. Penciclovir was significantly more active than acyclovir against three strains of HSV-1 and three strains of HSV-2 at 1 mg/L (P = 0.009), 3 mg/L (P < 0.001) and 10 mg/L (P = 0.001). Further comparisons between the compounds were made in MRC-5 cells infected with HSV-1 strain SC16 using four different antiviral assays namely, the 24 h virus yield reduction assay, plaque reduction assay, viral antigen inhibition assay, and a viral DNA inhibition assay, to determine the relative merits of each. Penciclovir and acyclovir shared similar activities in the plaque reduction assay (with 50% effective concentrations, EC50, being 0.8 and 0.6 mg/L, respectively) and in the viral antigen inhibition assay (EC50s. 0.6 and 0.7 mg/L, respectively). The EC50 of penciclovir in the 24 h viral DNA inhibition assay was 0.01 mg/L compared with 0.06 mg/L of acyclovir. In the 24 h virus yield reduction assay in which MRC-5 cells were infected with 0.3 pfu/cell, penciclovir was more active than acyclovir with 99% effective concentrations of 0.6 mg/L and 1.1 mg/L, respectively. The activity of penciclovir in the 24 h virus yield reduction and antigen inhibition assays was inversely related to the multiplicity of infection, whereas this had considerably less effect on the inhibition of viral DNA synthesis. These results suggest that famciclovir, which is the oral form of penciclovir, will be at least as effective as acyclovir in treating infections caused by HSV-1 and HSV-2. PMID- 8707741 TI - Retention of antibacterial activity and bacterial colonization of antiseptic bonded central venous catheters. AB - We determined how long antiseptic impregnation with silver sulphadiazine and chlorhexidine (SCC) on polyurethane central venous double- or triple-lumen catheters is retained in vivo. A total of 116 antiseptic catheters were tested for antibacterial activity in an in-vitro bioassay after various periods of iv catheterization. Segments from the subcutaneous (sc) and intravenous (iv) portions of the catheters were cultured. The results of test antiseptic catheters were compared with those from 117 noncoated control (c) catheters. Retention of antibacterial activity followed an exponential curve and lasted for up to 520 h after catheter insertion. Significant differences (P = 0.0001) between SSC and C catheters were noticed with regard to the quantitative level of bacterial colonization (SSC-sc 87 +/- 34 vs C-sc 584 +/- 122; SSC-iv 52 +/- 17 vs C-iv 286 +/- 57; all values are given as mean cfu +/- S.E.M.), and the frequency of bacterial colonization (SSC-sc 20.7% vs C-sc 38.5%, P = 0.0047; SSC-iv 18.1% vs C iv 30.8%, P = 0.0361). There was no significant difference between the incidence of catheter-related bacteraemia in the test (n = 0) and control groups (n = 3) (P = 0.2573). Further prospective studies are required to delineate the role of antiseptic catheters in preventing catheter-related infections. PMID- 8707742 TI - Treatment of experimental endocarditis caused by multidrug resistant Enterococcus faecium with ramoplanin and penicillin. AB - Antibiotic resistant strains of enterococci are being isolated with increasing frequency. Effective treatment of infections caused by Enterococcus faecium resistant to ampicillin, vancomycin and aminoglycosides has not been established. We studied the activity of ramoplanin, a new lipoglycopeptide antibiotic, against two strains of multidrug resistant E. faecium. In time kill studies, ramoplanin was bactericidal against both strains, but not in the presence of 50% serum. The combination of ramoplanin and penicillin was bactericidal even in the presence of serum. In rabbits with experimental endocarditis neither penicillin nor ramoplanin significantly reduced vegetation colony counts when given alone, although ramoplanin significantly reduced spleen and kidney bacterial counts of both strains. The combination of ramoplanin plus penicillin resulted in a significant reduction of vegetation bacterial counts (-3.2 and -3.7 log10 cfu/g for strains VA3 and MMC3, respectively, P < 0.01). All spleen cultures and 9 out of 10 kidney cultures from each strain were sterile following combination therapy. While ramoplanin will not be available for parenteral therapy, further research into the development of other lipoglycopeptide antibiotics is warranted. PMID- 8707743 TI - Distribution of cefdinir, a third generation cephalosporin antibiotic, in serum and pulmonary compartments. AB - The distribution of a new cephalosporin, cefdinir, in serum, epithelial lining fluid (ELF), bronchial mucosa and alveolar macrophages was studied in 17 adults following a single oral dose of 300 or 600 mg of cefdinir; tissue samples being obtained by diagnostic bronchoscopy approximately 4 h after this dose. Mucosal biopsies were taken, alveolar macrophages harvested by lavage, and ELF volume derived from urea concentrations in bronchial lavage fluid and blood. A microbiological assay for cefdinir was performed in serum, bronchial mucosa, ELF and alveolar macrophages. In patients taking 300 mg of cefdinir, the median concentrations of cefdinir were 2.00 mg/L (range 1.40-8.00) in serum, 0.78 mg/L (range 0-1.33) in bronchial mucosa, and 0.29 mg/L (range 0-4.73) in ELF. In patients taking 600 mg, the median concentrations were 4.20 mg/L (range 3.05 6.40) in serum, 1.14 mg/L (range 0-1.92) in bronchial mucosa, and 0.49 mg/L (range 0-0.59) in ELF. Cefdinir did not penetrate macrophages. PMID- 8707744 TI - Outer membrane permeability of non-typable Haemophilus influenzae. AB - The permeability to cephaloridine was studied in five Haemophilus influenzae strains (four non-typable and one type b) using the Zimmermann and Rosselet method. The beta-lactamase activity was due to a plasmid-encoded TEM-1 enzyme. High permeability coefficients were measured in all strains examined. No great differences in permeability coefficients were found, even between strains with marked differences in OMP electrophoretic profiles. PMID- 8707745 TI - Production of metal dependent beta-lactamases by clinical strains of Bacteroides fragilis isolated before 1987. AB - Five imipenem resistant clinical isolates of Bacteroides fragilis isolated before 1987, were examined to determine if they produced metallo-beta-lactamases. The beta-lactamases produced by the clinical isolates all focused as doublet bands at pl 4.8/4.9, characteristic of the B. fragilis CfiA type metallo-beta-lactamase. Each enzyme had a similar substrate profile and were inhibited by EDTA and activated with zinc sulphate. The sequence of the metallo-beta-lactamase gene from B. fragilis ED262 was determined and confirmed to be a CfiA type beta lactamase. Consequently, the five isolates examined probably produce a CfiA type beta-lactamase, suggesting that metallo-beta-lactamases were present before the widespread use of carbapenems. PMID- 8707746 TI - Mutations of the gyrA gene of clinical isolates of Salmonella typhimurium and three other Salmonella species leading to decreased susceptibilities to 4 quinolone drugs. AB - The region of the gyrA gene encoding nucleotides 72 to 557 in ten Salmonella typhimurium clinical isolates received from Vellore, India and six British clinical isolates of a number of Salmonella species has been sequenced. Those exhibiting a decreased susceptibility to 4-quinolone drugs (MICs of ciprofloxacin ranging from 0.128 mg/L to 1.0 mg/L) have been shown to possess a number of different mutations in this region of the gene (Asp-87 to Gly, Asp-87 to Asn and Ser-83 to Phe). The mutations Asp-87 to Gly and Ser-83 to Phe/Asp-87 to Asn have yet to be reported occurring in S. typhimurium. PMID- 8707747 TI - In-vitro activity of 3-azinomethyl-rifamycin (SPA-S-565) against Chlamydia trachomatis. AB - The in-vitro activity of a 3-azinomethyl-rifamycin (SPA-S-565) against Chlamydia trachomatis was compared to that of rifampicin and of erythromycin. The compound showed excellent activity for standard strains as well as isolates from patients with sexually transmitted diseases, being more active than the other drugs tested. SPA-S-565 also showed a low frequency of emergence of resistance. Passage of standard strains at sub-inhibitory concentrations caused an increase in MIC values of rifampicin while those for SPA-S-565 remained unchanged. PMID- 8707748 TI - Comparative in-vitro activity of SCH 27899, a novel everninomicin, and vancomycin. AB - The antibacterial activity of SCH 27899, was compared with vancomycin in vitro against 210 recent clinical isolates representing eight major genera of Gram positive organisms. MIC90 values for everninomicin were consistently better than those of vancomycin for all Gram-positive bacteria tested. Although everninomicin was less bactericidal than vancomycin against staphylococci, it was inhibitory against vancomycin-resistant enterococci. This novel antibiotic may, therefore, provide a clinical alternative to penicillin and vancomycin for therapy of Gram positive infections. PMID- 8707749 TI - In-vitro susceptibility of Streptococcus pneumoniae to the d- and l-isomers of ofloxacin: interpretive criteria and quality control limits. AB - The l-isomer of ofloxacin (levofloxacin) was twice as active as ofloxacin and the d-isomer was inactive against 654 Streptococcus pneumoniae isolates. For disc susceptibility tests of pneumococci, 5 micrograms levofloxacin discs can be used with interpretive criteria of < or = 12 mm for resistant (MIC > or = 8.0 mg/L) and > or = 16 mm for susceptible (MIC < or = 2.0 mg/L). For quality control of levofloxacin tests, S. pneumoniae ATCC 49619 should give zones 20-25 mm in diameter and MICs 0.5-2.0 mg/L. PMID- 8707750 TI - In-vitro antimicrobial susceptibility of the "Streptococcus milleri' group (Streptococcus anginosus, Streptococcus constellatus and Streptococcus intermedius). AB - A collection of 423 clinical "Streptococcus milleri' strains was speciated and tested for susceptibilities to twelve antibiotics. Only 1.4% of the strains were of intermediate susceptibility to penicillin. None of the strains exhibited high level resistance to gentamicin. Strains resistant to erythromycin, roxithromycin and clindamycin were found with a frequency of 2.6%, 2.4% and 2.4% respectively. Doxycycline resistance was found in 5.7% of the strains and occurred most frequently in Streptococcus anginosus. All the strains were susceptible to cefotaxime, vancomycin and teicoplanin. The results of this study do not indicate changing antibiotic resistance in strains of the "S. milleri' group, but local differences in antibiotic susceptibilities may occur. PMID- 8707751 TI - Killing of Streptococcus sanguis in biofilms using a light-activated antimicrobial agent. AB - The aim of this study was to determine whether Streptococcus sanguis, when in a biofilm, could be killed using a light-activated antimicrobial agent. Biofilms were grown on hydroxyapatite, irradiated with up to 12.2 J of light from a gallium aluminium arsenide laser in the presence of aluminium disulphonated phthalocyanine (AlPcS2) and survivors enumerated. No significant decrease in the viable count was found when either the AlPcS2 or the laser light was used alone. There was a light dose related decrease in the viable counts of irradiated AlPcS2 treated biofilms. No viable streptococci were detectable following irradiation with 12.2 J of laser light. PMID- 8707752 TI - Lack of interaction between valaciclovir, the L-valyl ester of acyclovir, and Maalox antacid. AB - Valaciclovir is rapidly and extensively converted to acyclovir. In this study we investigated the potential interaction between oral valaciclovir and Maalox. On each of three occasions 18 healthy volunteers received a single oral dose of 1000 mg valaciclovir, or 30 mL Maalox 65 min after valaciclovir administration, or 30 mL Maalox 30 min before valaciclovir. Acyclovir plasma concentrations and pharmacokinetic parameters were not significantly affected by administration of Maalox before or after valaciclovir. Therefore, there is no need for restriction of valaciclovir dosing in patients receiving antacid medication. PMID- 8707753 TI - Does the use in animals of antimicrobial agents, including glycopeptide antibiotics, influence the efficacy of antimicrobial therapy in humans? PMID- 8707754 TI - Photodegradation of fluoroquinolones using a continuous-flow photochemical reaction unit. PMID- 8707755 TI - Sensitivity of L-2 type beta-lactamases from Stenotrophomonas maltophilia to serine active site beta-lactamase inhibitors. PMID- 8707756 TI - The effect of betaine accumulation by Escherichia coli K-12 on aminoglycoside activity. PMID- 8707757 TI - Therapy-related myeloid leukemia. AB - One of the most serious possible consequences of cancer therapy is the development of a second cancer, especially leukemia. Several distinct subsets of therapy-related leukemia can be distinguished currently. These include classic therapy-related myeloid leukemia, leukemia that follows treatment with agents that inhibit topoisomerase II, acute lymphoblastic leukemia, and leukemias with 21q22 rearrangements or inv(16) or t(15;17). These types of leukemia are discussed in detail in this article. PMID- 8707759 TI - Anemia of cancer. AB - Cancer patients frequently develop anemia, due either to the cancer itself or to the effects of cancer-related therapy. Recent years have brought insights into both the pathogenesis of the anemia of cancer and the extent to which erythropoietin regulation participates in this process. Although transfusion therapy was the mainstay of therapy for symptomatic anemia in the past, clinical trials have demonstrated that recombinant human erythropoietin can alleviate both anemia and transfusion requirements in many cancer patients and may prove to have an important role in the treatment of cancer-related anemia in the future. PMID- 8707758 TI - Bone marrow metastases. AB - This article discusses the clinical significance of bone marrow metastases and the current methods being used to detect tumor cells in marrow. The strategies being investigated for eradicating cancer cells from marrow in patients receiving hematopoietic cell autografts also are reviewed. PMID- 8707760 TI - Hemolytic disorders associated with cancer. AB - Three principal environmental causes of hemolytic anemia in malignancy have been identified: (1) hemolysis mediated by auto-antibodies to red cells; (2) hemolysis due to microangiopathic disorders; and (3) chemotherapy-induced red cell destruction. These three environmental stressors occur rarely in cancer patients, and they form the subject of this review. PMID- 8707762 TI - Transfusion and stem cell support in cancer treatment. AB - Intensification of therapeutic regimens, improved patient survival, and advances in cytokine and cellular therapies have led to increasingly complex requirements for transfusion and stem cell support in cancer treatment. This article focuses on current and evolving issues in red blood cell, platelet, and granulocyte transfusion support, as well as measures to avoid increasingly important complications of transfusion therapy, such as alloimmunization, graft-versus-host disease, cytomegalovirus infection, and immunomodulation. Issues concerning current applications of hematopoietic stem cell transplantation and future prospects also are discussed. PMID- 8707761 TI - Chemotherapy and neutropenia. AB - Myelosuppression is the most common toxicity associated with the administration of dose-intensive cytotoxic chemotherapy. The basic understanding of neutrophil biology and the physiology of chemotherapy-induced neutropenia has advanced tremendously in the past 2 decades. Concordantly, the ability to reduce the morbidity associated with neutropenia has improved. Adjunctive cytokine and progenitor cell support of hematologic recovery after myelosuppressive therapy have proved to be models of translational research and have led to novel therapeutic initiatives for patients with cancer and hematologic malignancies. In this article, fundamental aspects of neutrophil production are discussed, and the clinical development of hematopoietic cytokines active on cells of the leukocyte lineages is presented. PMID- 8707763 TI - The thrombocytopenia of cancer. Prospects for effective cytokine therapy. AB - The previous 10 years have witnessed the development of increasing needs for platelet transfusion in support of aggressive therapies of malignancy. Despite gains in our understanding of platelet preparation, storage, and transfusion, alternative therapies are clearly desirable. During the late 1980s at least six distinct cytokines that display effects on megakaryocyte growth and differentiation-IL-3, KL, GM-CSF, IL-6, IL-11, and LIF- and a synthetic growth factor, PIXY 321, were cloned and characterized. Although none of these cytokines fulfill all of the physiologic roles of thrombopoietin, in its absence several have undergone extensive preclinical and preliminary clinical testing. Of these, IL-11 and PIXY 321 hold promise for clinical amelioration of thrombocytopenia in cancer patients. With the recent cloning of thrombopoietin and its promise in preclinical trials, the role of each of these recombinant proteins in clinical medicine is undergoing careful evaluation. As with erythropoietin and G-CSF before it, given its normal role in the regulation of platelet production, Tpo would appear to provide the greatest physiologic stimulus to platelet production in states of natural and iatrogenic marrow failure. Careful clinical trials of the agent are needed to determine whether the hormone will fulfill this promise. The following decade will most certainly see the resolution of many of the complications of thrombocytopenia and its transfusional support. PMID- 8707764 TI - Coagulation disorders in cancer. AB - Coagulation disorders are common in cancer patients. This article reviews the coagulation laboratory findings in these patients and the thromboembolic and hemorrhagic manifestations of malignancy. Among the many topics addressed are Trousseau's syndrome, disseminated intravascular coagulation, and acquired von Willebrand disease. Pathogenesis of the coagulation disorders and recommendations for treatment of various syndromes are discussed. PMID- 8707765 TI - Thrombotic microangiopathies associated with drugs and bone marrow transplantation. AB - This type of thrombotic microangiopathy more commonly resembles the hemolytic uremic syndrome (HUS) than thrombotic thrombocytopenic purpura (TTP). The syndrome has been associated with the use of cyclosporin, mitomycin C, combinations of other chemotherapeutic and immunosuppressive agents, and total body irradiation. Endothelial cell injury and von Willebrand factor may be involved in pathogenesis of the intravascular platelet aggregation and tissue (especially renal) ischemia and infarction that characterize the entity. The most effective therapy for thrombotic microangiopathy associated with drugs and bone marrow transplantation has not been determined. PMID- 8707766 TI - Hypercoagulability in cancer. AB - The association of cancer with a hypercoagulable state is documented by numerous clinical, biochemical, pathologic, and pharmacologic studies. This association is manifested clinically by an increased incidence of intravascular thrombotic events in cancer patients and by fibrin deposition in and around tumor beds. Thromboembolic disease is a major cause of morbidity and mortality in patients with malignancy. This article discusses the complex pathogenesis of this problem and the associated laboratory and clinical syndromes with recommendations on diagnosis and treatment. PMID- 8707767 TI - Statistical considerations and modeling of clinical utility of tumor markers. AB - The proper assessment of potential tumor markers in the screening, diagnosis, prognosis, and monitoring of neoplastic diseases depends on careful attention to general principles of good design and analysis of scientific studies, as well as to some specific statistical principles to avoid erroneous conclusions. Published reports on tumor markers should be evaluated critically with these principles in mind. It is well known that statistical problems exist in the medical literature despite the peer review process. In addition, attention to these principles in the design of studies of tumor markers can help ensure their accuracy and validity and avoid the common pitfalls. PMID- 8707768 TI - Markers of risk for human malignancies. AB - Markers of risk of human cancer fall into many categories. This article is limited to heritable genetic markers of specific familial cancer syndromes as examples of measurable markers of cancer risk. Dominantly inherited cancer susceptibility genes confer risks of cancer often greater than any known environmental exposure. PMID- 8707769 TI - Tumor markers for breast cancer. Current utilities and future prospects. AB - Tumor-associated markers have potential utility in identification, screening, prognosis, detection, or monitoring breast cancer. Of the available markers, those with the greatest promise in 1994 include the yet-to-be-cloned BrCa1, the p53 tumor suppressor gene, tissue-associated prognostic factors such as HER 2/neu, cathepsin-D, and indicators of angiogenesis, and circulating tumor markers that provide an indication of clinical course, such as CA15-3 and CEA. Unfortunately, the precise clinical utilities of all of these markers remain imprecise. It is especially important that the relative independence of the markers in relation to other available markers be determined so as to avoid the unnecessary cost and expense of redundancy. Moreover, it is important that the clinician be aware of the limitations in both sensitivity and specificity of each marker so as not to over- or underinterpret the predictive value of any test. With these caveats in mind, judicial application of germline, tissue, and soluble tumor markers can improve clinical care of patients at risk for and with breast cancer. PMID- 8707770 TI - Use of tumor markers in lung cancer. AB - Biomarkers relevant to lung cancer can be classified into three broad categories: serum markers, tissue markers, and sputum markers. The predominant contribution of serum biomarkers will be in documenting disseminated disease and response to therapy. The most promising role for tissue biomarkers, at least in non-small cell lung cancer, is for the determination of prognosis in resectable patients. Sputum biomarkers offer considerable promise as early detection tools, which may enhance the sensitivity and specificity of screening when used in conjunction with periodic screening chest radiographs. PMID- 8707771 TI - The use of serologic tumor markers in gastrointestinal malignancies. AB - The tumor markers CEA, CA 19-9, and AFP have a distinct and useful role to play in the management of gastrointestinal malignancies. These are clearly not ideal tumor markers. They lack day-to-day reproducibility and sufficient sensitivity and specificity to be useful as general screening tools. Taken as isolated values, they are lacking in precision. However, used in the appropriate context, with an understanding of their associated biology, these tumor markers help to manage patients and might reduce the economic burden of decision making. As we develop a better understanding of the process of neoplastic transformation and progression, new tumor markers will become available. Markers may, in the future, be used to predict responses to particular therapies or overall prognosis. Some will be molecular probes, such as the K-ras or FAP gene mutations, and others may relate to alterations in regulated processes, such as increased expression of a key enzyme in DNA synthesis, thymidylate synthase. PMID- 8707772 TI - The prostate specific antigen. Its use as a tumor marker for prostate cancer. AB - The prostate specific antigen (PSA) has had an enormous impact on the diagnosis and management of prostate cancer. Use of the PSA has strikingly increased the diagnosis of prostate cancer since its introduction and widespread application in clinical practice. This article examines the assay characteristics of PSA, its utility in early detection of prostate cancer, and its use in facilitating treatment decisions. PMID- 8707773 TI - The use of tumor markers in germ cell malignancies. AB - Serum tumor markers, including alphafetoprotein and human chorionic gonadotropin, are critical to the choice of therapy and monitoring of patients with germ cell tumors. i(12p) is a specific chromosomal marker for germ cell tumors that is diagnostically useful, of possible prognostic importance, and detectable by both conventional and molecular cytogenetic techniques. A variety of other molecular genetic markers are under study, in order to contribute to the understanding of normal and aberrant differentiation and to understand their relevance as tumor markers. PMID- 8707774 TI - Compliance with asthma therapy: a case for shared responsibility. PMID- 8707775 TI - Medication compliance and asthma: overlooking the trees because of the forest. AB - A number of reviews have described medication compliance and asthma. The consensus emerging from the analyses and discussion of the topic is that compliance to treatment regimens is generally poor. The current article describes several trees overlooked in the forest of data generated about compliance and asthma. The paper focuses on neglected issues concerning determinants of compliance, assessment, and changing noncompliant behavior. Based on available data, the conclusion is that it is time for health care personnel and patients to share the blame for the lack of medication compliance in asthma. PMID- 8707776 TI - Increased hypodense eosinophils after activation with PAF-acether and calcium ionophore in asthmatic subjects. AB - Eosinophils play a major role in the pathogenesis of bronchial asthma. In this study, we examined the density characteristics of blood eosinophils from 9 normal healthy individuals and 9 allergic asthmatic patients. Furthermore, the effect of platelet-activating factor, a potent mediator of inflammation, and calcium ionophore, A23187, on the density of normodense eosinophils (density > 1.085 g/ml) has also been examined. Initially, asthmatic patients had 27.0 +/- 1.1% eosinophils of lighter density (density < or = 1.081 g/ml), significantly greater than that in the normal individuals (7.5 +/- 0.5%). After exposure to platelet activating factor (1 microM) or calcium ionophore (A23187, 1 microgram/ml), the normodense eosinophils switched to hypodense in both groups: 16.7 +/- 2.1% and 54.2 +/- 3.7%, respectively, in normal individuals, and 30.6 +/- 5.7% and 77.4 +/ 2.3%, respectively, in asthmatic patients. These data demonstrated that a certain percentage of normodense eosinophils from asthmatics and normal subjects switched to hypodense after activation with platelet-activating factor or calcium ionophore. Furthermore, eosinophils from asthmatics switched to a greater degree than in normal subjects, suggesting that the normodense eosinophils in asthmatics become primed probably by endogenously released mediators. PMID- 8707777 TI - The capacity of children with and without asthma to detect external resistive loads on breathing. AB - Clinical observations and research with adults consistently showed that subjective symptoms of asthma poorly reflect actual airway obstruction. The lack of accurate symptom perception poses a problem for medication and management of asthma. The accuracy of airflow detection was studied in 46 children with and 46 without asthma (aged 7-18 years). They breathed through a facemask and responded to load stimuli of different intensity. Sessions consisted of 10 blocks of 5 min, each with 10 stimuli presented. Experiment 1: Loads of increasing intensity presented to 36 children with and 36 without asthma. Seven asthmatics had a reliable detection threshold (just noticeable difference, jnd) analogous to approximately equal to 64% fall in forced expiratory volume in 1 sec (FEV1). Ten normal controls had a jnd of approximately 39% fall. Experiment 2: Loads randomly presented to 10 children with and 10 without asthma. Four asthmatics had a jnd of approximately equal to 64% fall in FEV1. Six normal controls had a jnd of approximately equal to 39% fall. The results demonstrated that children generally were poorly perceiving load stimuli and that asthmatics were less accurate. PMID- 8707778 TI - Hypopigmented skin lesions associated with atopic dermatitis in asthma. AB - The association of vitiligo and other skin pigmentary disorders has been well shown with autoimmune diseases, i.e., systemic lupus erythematosus, rheumatoid arthritis. Furthermore, in certain diseases such as diabetes mellitus and adrenal insufficiency the incidence of pigmentation disorders is higher than in the general population. A total of 53 male and 46 female patients with a diagnosis of asthma, based on clinical findings and pulmonary function tests (PFT), were thoroughly examined for the presence of hypopigmented skin lesions and compared with 100 nonasthmatic controls with a similar demographic distribution. The patients were questioned regarding the history of steroid treatment in the past and present and subdivided into five groups based on the duration of therapy. Adrenal gland function was tested by 8:00 A.M. serum cortisol concentration in the group of patients with a history of steroid use as well as in some patients without this history. These patients were also tested for absolute lymphocyte and eosinophil count. Hypopigmented lesions were noted in 36.4% of asthmatic patients and in 11% nonasthmatic controls (p-value < 0.01). There were no differences in the location, but the size of lesions per patient was larger in the asthmatic than the control group. Twenty of 55 patients who had a history of steroid treatment in the past had hypopigmented skin lesions as compared to 16 of 42 patients who had no history of steroid treatment. We conclude that there is a higher coincidence of hypopigmented skin lesions, mainly vitiligo-type in asthmatic patients, compared to the general population, which is not related to a history of steroid treatment or incidence of adrenal insufficiency in these patients. PMID- 8707779 TI - Education of parents of infants and very young children with asthma: a developmental evaluation of the Wee Wheezers program. AB - A randomized control trial of the Wee Wheezers asthma education program was conducted with 76 children < 7 years of age, 31% of whom were on a medication regimen consistent with mild, 51% with moderate, and 18% with moderately severe/severe asthma. Treatment children showed improved morbidity at 3-month follow-up relative to the changes in the controls: increased symptom-free days in the preceding 2 weeks (mean change of +2.2 vs. -2.6 in the controls; p = .004) and month (+2.0 vs. -3.8; p < .02), fewer nights of parental sleep interruption in a typical week (+0.7 vs. +1.8; p < or = .05), and a trend toward fewer asthma sick days (-0.2 vs +0.7; p = ns). These improvements were accompanied by significantly better parental asthma management compared with controls (more consistent use of preventive medications, p < or = .01; early symptom intervention, [corrected] p < or = .05) and trends toward more restrictions on smoking in the home (p < .07) and decreased parental confusion about asthma treatment (p < .11). This study provides evidence that a multisession program of asthma education for parents can improve parental asthma management and clinical outcomes in very young children and provides information on the validity and sensitivity of various asthma outcome measures in this age group. PMID- 8707780 TI - Neonatal characteristics as risk factors for preschool asthma. AB - Childhood asthma usually begins early in life. Neonatal characteristics are reportedly predictive of symptom onset. This investigation utilized data from a provincial health organization to evaluate the effect of several birth characteristics on asthma incidence and hospitalization for asthma during age 0 4. Using logistic regression, the odds ratios (OR) for the following variables indicate a significant (p < 0.05) association with physician-diagnosed preschool asthma: male gender (OR = 1.72), birthweight < 1500 g (OR = 2.11), prematurity (OR = 1.34), respiratory distress syndrome (RDS) in the presence (OR = 2.95) or absence (OR = 1.61) of bronchopulmonary dysplasia (BPD), and transient tachypnea of the newborn (TTN; OR = 1.36). Male gender (OR = 1.91), birthweight < 1500 g (OR = 2.56), RDS with and without BPD (OR = 3.35 and 2.50, respectively), TTN (OR = 2.08), and severe birth asphyxia (OR = 1.94) showed an important association with hospitalization due to asthma. Neonatal characteristics are important determinants for the risk of preschool asthma, even after mutual adjustment. PMID- 8707781 TI - A 12-week dose-ranging study of fluticasone propionate powder in the treatment of asthma. AB - Fluticasone propionate (FP) administered via metered-dose inhaler is a potent corticosteroid effective in the treatment of asthma. To evaluate the efficacy and safety of FP powder administered via a breath-activated inhaler (Diskhaler), a multicenter, double-blind, randomized, placebo-controlled, parallel-group study was conducted in adolescent and adult patients (n = 331) with mild-to-moderate asthma previously treated with beta 2-agonist therapy alone. Patients received FP powder 50, 100, or 250 micrograms or placebo twice daily for 12 weeks. FP-treated patients compared with placebo-treated patients had significantly (p < 0.001) greater improvements in morning predose forced expiratory volume in 1 sec (21-22% increase vs. 9%). Improvement in morning peak flow rate were also significantly (p < 0.001) greater with FP than with placebo (8-10% increase vs. 2% increase). There was also a significant overall treatment difference in the frequency of inhaled albuterol use (p < 0.001) and number of nighttime awakenings due to asthma (p = 0.005). There were no statistically significant difference among the FP treatment groups in any outcome measure. Physicians' global assessments also indicated significant (p < 0.001) differences in efficacy, with 67-74% of FP treated patients rated as having "effective" or "very effective" treatment compared with 41% of placebo-treated patients. Significant beneficial effects of FP were observed in lung function and diary card parameters after just 1 week of treatment. Adverse events were similar across treatment groups and primarily related to local irritation. Effect on hypothalamic-pituitary-adrenal axis function was minimal. In summary, all three dosages of inhaled FP powder were well tolerated and improved various asthma-related variables. Improvements in pulmonary function, beyond those achieved with beta 2-agonist therapy alone, were maintained for the duration of the 12-week study. PMID- 8707782 TI - Regulation of class I and class II myosins by heavy chain phosphorylation. PMID- 8707783 TI - Stress and strain in the flight muscles as constraints on the evolution of flying animals. AB - The minimum mechanical power needed for an animal to maintain level flight can be estimated, as can the wingbeat frequency, from measurements of the animal's mass, wing span and wing area, and of the strength of gravity and the air density. Dividing the power by the frequency gives the work done per cycle, and dividing this by the muscle mass gives the specific work, meaning the work done in each contraction by unit mass of muscle. This in turn is the product of the average stress during shortening and the strain, divided by the muscle density. The minimum specific work for level flight is strongly size dependent. To account for even minimum performance in the largest species known to be capable of prolonged, aerobic flight (whooper swan), the specific work of the myofibrils needs to be 57 J kg-1, which could be achieved, for example, by a stress of 240 kN m-2 combined with a strain of 0.25. The upper limits of stress and strain for sustained exercise are not known, but are not likely to be much higher than these figures. Much larger birds, such as the Miocene fossil Argentavis, would require improbably high values of stress and strain for level flight, unless the air density were much higher in Miocene times than at present, and/or the strength of gravity were much less. Birds of small and medium size have more than the minimum amount of muscle required for level flight. This opens a wide range of possibilities for different species to be specialised for different types of activity. The potential diversity for evolution in large species is less than for medium-sized or small ones, and dwindles to zero above a body mass of about 14 kg. There is also a strong positive trend in the aerobic scope, from about 3 in small, long-winged passerines such as swallows, to 47 in the whooper swan. PMID- 8707784 TI - A functional knee brace alters joint torque and power patterns during walking and running. AB - Individuals with anterior cruciate ligament (ACL) injury use greater extensor torques at the hip and ankle and lower extensor torques and joint power at the knee during gait compared to healthy subjects. These adaptations may be mediated by (1) altered neuromuscular strategies due to the injury, (2) training effects produced by rehabilitation protocols, and (3) training effects due to the functional knee brace (FKB) used during rehabilitation. The purpose of the study was to test the hypothesis that a FKB can cause individuals to walk and run with the torque and power patterns observed in rehabilitated ACL-injured individuals. Ten healthy subjects were tested walking and running with and without a FKB. Kinematic and ground reaction data were collected and combined with inverse dynamics to estimate the joint torques and powers. Data were analyzed with a two way repeated measures ANOVA (gait vs knee condition). In walking, the hip and ankle extensor torques were 14.3% (p < 0.038) and 5.1% (p < 0.003) greater with FKB. In running, the hip extensor torque was 17.0% greater with FKB (p < 0.023). Knee torque was not different between conditions. In walking, the work performed at the hip and knee were 11.6% greater (p < 0.013) and 17.7% lower with FKB (p < 0.025), respectively. Results supported the hypothesis and it was concluded that a FKB may be one causative factor in the development of the unique joint torque and power patterns seen in ACL-injured gait. PMID- 8707785 TI - Features of human jaw design which maximize the bite force. AB - A linear programming model, operating in a three-dimensional coordinate system, was written to study the effect on unilateral maximum bite forces (MBFs) of small changes in the design of human jaws. Under some conditions the MBF was larger on a more anterior tooth than a more posterior tooth for a given bite direction. The MBF was increased (i) as the bite force was tilted forward, (ii) as the jaw was opened if its direction was kept perpendicular to the occlusal plane of the lower jaw, (iii) by placing teeth nearer the midline, (iv) by raising the height of the teeth for forward bite forces, and (v) by tilting forward the articular surface of the condyle (equivalent to creating an articular eminence). Increasing the distance between the condyles spread the load more evenly between the two joints. The articular eminence was the only design feature which seemed to be clearly related to increasing the MBF. PMID- 8707786 TI - Tethering affects the mechanics of coronary capillaries. AB - Coronary capillaries are extensively tethered to adjacent myocytes by collagen fibers. The influence of this tethering in the beating heart is studied by structural mechanics as applied to the specific morphology of the capillary myocyte system. The results show considerable effects of the tethering collagen fibers on the capillary deformation, especially during systole and in the deeper myocardial layers. The tethering fibers prevent total systolic collapse, being taut during systole but partially slack during diastole, in agreement with reported observations. At the deeper wall layers, the systolic/diastolic differences in capillary cross-sectional area are predicted to be more pronounced: about 30 and 50% area reduction in arterial and venous ends, respectively, compared with 10 and 20% increase of area in the subepicardial vessels. These predictions comply well with published, experimental data. A parametric investigation shows a variable effect of the capillary-myocyte distance on the dynamics of the capillary area, while the stiffnesses of both the fibers and wall membrane, and the extent of transmural transmission of intramyocardial pressure, have both considerable quantitative effects. These effects are found to be region dependent and vary along the capillary length and from diastole to systole. The results indicate that capillary tethering to the myocardial tissue has significant effect on its mechanics. Tethering should, therefore, be considered in analyzing the dynamics of coronary flow. PMID- 8707788 TI - Normalization of temporal-distance parameters in pediatric gait. AB - Several techniques for the normalization of temporal-distance parameters in pediatric gait are given. The resulting normalized data can be used to compare (or discriminate) individuals or groups of individuals without the effect of variables such as age and height. The normalization with respect to a reference data set or with respect to the data set itself can be accommodated. Three novel techniques for normalization of gait data have been given: offset, decorrelation and detrending. For normalization of stride length with respect to height, all three are superior to the commonly accepted technique of dividing by the height. The offset technique has obscure units and will have a residual correlation. The decorrelation technique also has obscure units but will have zero correlation provided it is being normalized using a linear model (or piecewise linear model) fitted to the data in a least-squares sense. The detrending technique will also result in a zero correlation if piecewise linear or polynomial models, fitted to the data in a least-squares sense, are used. The detrending technique is the most useful of the three techniques proposed as it will also generate the same units as the original data set and can be easily scaled so that its magnitude is also comparable with the original data. Both the decorrelation and detrending techniques can be used simultaneously to normalize data with respect to two or more variables. PMID- 8707787 TI - A three-dimensional, time-dependent analysis of flow through a bileaflet mechanical heart valve: comparison of experimental and numerical results. AB - The flow through a bileaflet mechanical heart valve during the first half of systole was predicted using computational fluid dynamics (CFD). A three dimensional model of the geometry of the ventricle, valve, sinus and aorta was developed. Flow through the valve was assumed to be Newtonian and laminar. The peak systolic Reynolds number was 1500 based on the aortic radius and the mean aortic velocity. Flow visualisation and laser Doppler anemometry (LDA) experiments were performed and the results were compared to the CFD model. Good agreement between the LDA measurements and CFD predictions was found in the jets through the major orifices of the valve. The global flow fields predicted by the CFD showed reasonable agreement with the flow visualisation. A starting vortex was shed from the valve leaflets of the CarboMedics valve and the prototype valve. As systole progressed the two major orifice jets were directed towards the aortic wall and a weaker central jet was seen in both the experimental and CFD models. Large vortices were present on either side of the central orifice jet in the sinus area of both models. The three-dimensional time-dependent CFD model was considered to give a reasonable indication of the dominant flow patterns downstream of the bileaflet heart valve and has the potential to be an extremely useful tool to analyse the different designs of existing and future bileaflet valves. PMID- 8707789 TI - Stability of muscle fibers on the descending limb of the force-length relation. A theoretical consideration. AB - For the past 40 years it has generally been accepted that the descending limb of the force-length (F-L) relation of muscle fibers is unstable; strong sarcomeres are thought to shorten onto the ascending limb of the F-L relation at the expense of weak sarcomeres which are stretched beyond the thin-thick myofilament overlap. This unstable behavior has not been demonstrated by direct observation but has been inferred indirectly, based on the negative slope of the F-L relation. Intuitively, the idea that the sarcomere length within skeletal muscle is unstable at any length within the normal working range would not be advantageous. Therefore, the purpose of this study was to identify analytically the mechanical conditions for which sarcomeres in a muscle fiber are stable. It was found that the mechanical conditions required for sarcomere and fiber stability could be associated with known properties of skeletal muscle fibers. Furthermore, it could be demonstrated that a fiber composed of a large number of sarcomeres can be stable, and still exhibit an apparently unstable (negatively sloped) descending limb of the F-L curve. PMID- 8707790 TI - Theoretical study of dynamics of arterial wall remodeling in response to changes in blood pressure. AB - The dynamics of arterial wall remodeling was studied on the basis of a phenomenological mathematical model. Sustained hypertension was simulated by a step increase in blood pressure. Remodeling rate equations were postulated for the evolution of the geometrical dimensions that characterize the zero stress state of the artery. The driving stimuli are the deviations of the extreme values of the circumferential stretch ratios and the average stress from their values at the normotensive state. Arterial wall was considered to be a thick-walled tube made of nonlinear elastic incompressible material. Results showed that thickness increases montonically with time whereas the opening angle exhibits a biphasic pattern. Geometric characteristics reach asymptotically a new homeostatic steady state, in which the stress and strain distribution is practically identical with the distribution under normotensive conditions. The model predictions are in good agreement with published experimental findings. PMID- 8707791 TI - Correcting trunk muscle geometry obtained from MRI and CT scans of supine postures for use in standing postures. AB - Many studies have reported the geometry of the trunk muscles for use in models to predict spine loads. However, all have reported data collected from subjects positioned in a prone or supine posture which is required by CT or MRI scan techniques. This study compared the abdominal and extensor muscle moment arms obtained with muscles relaxed and activated and in both supine and upright postures. Both internal ultrasound measurements and external anthropometer measurements were collected. The anterior abdominal wall increased its flexor moment arm by approximately 30% when in a standing posture compared to the supine posture. The effect of an upright posture on the moment arm of erector spinae at the level of L3 was much smaller (3% increase in males and 12% increase in females). There was no moment arm change due to activating the muscles when compared to the relaxed condition for either posture. Modelers should be aware of the need to scale muscle moment arms obtained from MRI/CT scans in order to more accurately predict spine loads in upright postures. PMID- 8707792 TI - Strain or deformation rate dependent finite growth in soft tissues. AB - The particular portion of the mechanical loading history of a tissue that serves as stimulus for growth or remodeling of that tissue is presently unknown. A kinematic basis for the implementation of strain or rate of deformation as a growth stimulus is presented. It is shown here that a recently proposed continuum theory for finite volumetric growth in soft tissue may be extended to include strain and rate of deformation as growth stimuli. The basis of the presentation is the recognition of two different time scales in the remodeling process, one on the order of seconds associated with the loading, and one on the order of weeks or months associated with the growth. PMID- 8707793 TI - Methods for investigating the sensitivity of joint resultants to body segment parameter variations. AB - Two methods are presented for determining the sensitivity of joint resultants to variations in body segment parameters (BSPs) when solving inverse dynamics problems: the exact sampling method (ESM), and the approximate variational method (AVM). These methods are illustrated by applying them to a single segment in fixed-axis rotation. The results indicate that (a) the AVM provides first-order (linear) approximations to the total variations in joint resultants obtained when using the ESM, (b) the ESM should be used when BSP variations become large and higher-order terms in these variations can no longer be ignored as negligible in the BSP-dependent expansions for the joint resultants, (c) small percent variations in BSPs can propagate into considerably larger percent variations in the joint resultants obtained when solving inverse dynamics problems, and (d) if BSP variations are sufficiently small to allow nonlinear terms in these variations to be ignored as negligible, the AVM is much simpler and easier to use than the ESM. PMID- 8707794 TI - Stress-strain curve and Young's modulus of a collagen molecule as determined by the X-ray diffraction technique. AB - The purpose of the work described in this paper was to make a stress-strain curve for a collagen molecule and estimate Young's modulus of a molecule along the molecular axis. X-ray diffractometry was performed on bovine Achilles tendon in order to measure strain in the collagen molecule along the molecular axis as a response to a macroscopically applied force. By geometrical calculations and experiments, cross-sectional areas of a molecule and molecules in a tendon collagen fiber were determined. The applied force was translated to the stress and the stress-strain curve of the collagen molecule was constructed, which was found to be almost linear. Young's modulus of the molecule was determined to be slightly smaller than when determined by dynamic mechanical methods. The difference was considered to suggest the existence of a viscoelastic component within the molecule as well as the difference in the mechanical properties of collagen in different tissues. The expected viscoelasticity was speculated to be related to the hydrogen bond network in the collagen molecule. PMID- 8707795 TI - Static in situ calibration of force plates. AB - An in situ calibration protocol for ground-to-foot force measuring platforms is described. The methodology allows verification of the function of the force plate and allows accurate calibration for three force and moment channels. The effect of cross-sensitivity on recorded data is discussed along with the need for improvements in methodology to quantify this property. PMID- 8707796 TI - Instrumented implant for measuring tibiofemoral forces. AB - Experimental measurement of loads occurring in the human knee joint will allow validation of analytical models and provide data for the design of total knee implants. A customized transducer was developed to measure the dynamic tibiofemoral force and center of pressure after total knee arthroplasty. The transducer consists of a standard tibial component to which four uniaxial load cells and an additional tibial tray have been added. The transducer was calibrated using a loading device traceable to the National Institute of Standards and Technology (NIST). The transducer was accurate to within 1% in magnitude, 0.07 mm in medial/lateral location and 0.24 m in anterior/posterior location. PMID- 8707797 TI - Three-dimensional videography of swimming with panning periscopes. AB - The purpose of this paper is to describe a three-dimensional videography method with panning periscopes for reconstructing swimming techniques. Two panning periscope systems, developed in our laboratory, were used for the data collection. A control object (1 x 1.2 x 2)m3 with 72 markers was placed in seven consecutive locations along the (1.5 x 8.4 x 2) m3 calibration space. The position of the control object was recorded while panning the periscope systems, and for each of the seven locations the DLT camera parameters were determined for eight fields equally spaced through the panning motion. Each DLT camera parameter was then expressed as a function of the camera orientation by using a least squares, second-order, polynomial regression equation, so that the DLT camera parameters for any camera orientation could be predicted (Yu et al., J. Biomechanics 26, 741-751, 1993). The three-dimensional coordinates of desired body landmarks at an instant were determined from the digitized two-dimensional coordinates for two cameras and the predicted DLT camera parameters at that instant. The accuracy of the method was evaluated by (a) the resultant errors in computing three-dimensional coordinates of precalibrated static points, and (b) the errors in computing the length of a scale rod pulled through the calibration space. The mean resultant errors ranged from 8.34 to 16.44 mm for the above- and from 9.93 to 16.22 mm for the below-water control volumes. The mean error in computing lengths on the scale rod ranged from 3.32 to 5.83 mm for three 0.5 m lengths, and 9.97 mm for a 1.5 m length. The method produced acceptable results in the reconstruction of three-dimensional motions recorded from a large space above and below the water surface. PMID- 8707798 TI - A new method for theoretical analysis of static indentation test. AB - A new mathematical method was developed to study the indentation problem of an infinite elastic layer overlaid on a rigid foundation. Rigid, flat-ended cylindrical or spherical indenters are pressed onto the upper surface of the elastic layer causing a small deformation mode. Shear stresses between the indenter and the layer are assumed negligible and the layer is assumed to be either bonded or unbonded to the rigid foundation. The problem is equivalent to a mixed boundary-value problem of the theory of elasticity. Instead of using the Fredholm integral equation reported in the literature, the new approach obtained closed-form solutions through an infinite series. Convergence can be achieved using less than 10 terms of the series. PMID- 8707799 TI - A sliding-distance-coupled finite element formulation for polyethylene wear in total hip arthroplasty. AB - A three-dimensional, nonlinear contact finite element (FE) model of total hip replacement, linked to a sliding-distance-coupled wear algorithm, was used to study polyethylene wear rates for three different femoral head sizes. Hip resultant loads from a validated gait analysis model were used in the FE model to determine contact stress distributions on the polyethylene bearing surface, for 16 discrete instants of stance phase. Sliding distances of points on the femoral head surface were obtained from the corresponding flexion/extension kinematics. Wear rates were determined by a custom-written computer program that used a relationship that coupled contact stress, sliding distance, and a pin-on-disk determined wear coefficient. The wear rates computed by this formulation were well within clinically observed ranges for each component size. PMID- 8707800 TI - Segment inertial parameter evaluation in two anthropometric models by application of a dynamic linked segment model. AB - The estimation of segment inertial parameters (SIPs) is an important source of error in inverse dynamic analysis. In most individual cases SIPs are derived from extrapolation of known SIPs of a certain population through regression equations (proportional models). Another well-known method is the use of mathematical approximation of the shape of human body segments combined with estimations of segment densities (geometric models). In the current study five males and five females performed four different lifting movements in the sagittal plane. A full body linked segment model was applied twice to the same data set, once using a proportional and once using a geometric anthropometric model. As a full body linked segment model is an overdetermined system of equations, four equations could be formed to test the summed effect of SIP errors on the inverse dynamic analysis. The overall performance in terms of coefficients of correlation was better for the geometric model as compared to the proportional model. When a back lifting movement was performed, the equations indicated systematic errors in the proportional model. However, when a leg lifting movement was performed, the equations indicated systematic errors in the geometric model. Therefore, analyzing only one kind of movement does not suffice to draw conclusions with respect to the reliability of an anthropometric model. PMID- 8707801 TI - Angioscopy in the operating room. AB - Angioscopy has been used in the superficial femoral artery for many years, as well as in the control of vascular surgery procedures. It is more reliable than angiography, which often misses such features as intimal flaps, residual valves in "in situ" vein bypasses or persisting thrombus after embolectomy. As fluid irrigation allows visualization of the iliac arteries, and as smaller atraumatic fibres allow safe visualization of the tibial arteries, new fields are opened for the use of angioscopy in peripheral vascular disease. This technique has been proved to be useful: (1) as a diagnostic tool, (2) as a control device in new endovascular procedures (percutaneous angioplasty, atherectomy and stents), and (3) as a therapeutic tool (thromboembolectomy, simplification of bypass procedures). In a modern operating room, angioscopy does not replace angiography, but these two imaging techniques are complementary, as they help the surgeon to provide the patients with a less invasive vascular surgery. PMID- 8707802 TI - The surgeon as an endovascular interventionist. Why and how? AB - Catheter-based endovascular techniques have advanced significantly over the past three decades. It is only recently that surgeons have become more interested in these developments, and are now rapidly gaining in training and widening practice opportunities. Endovascular training and credentialing for the surgeon continue to be poorly-defined areas. Several pathways, albeit incomplete, are available for the incorporation of endoluminal/percutaneous skills into the surgical armamentarium. Ultimately, it can only be through established postgraduate residency and fellowship programs that the subspecialty of endovascular surgery can have a more universal applicability. The basic necessary catheter skills have been defined. Imaging requirements, both radiographic and endoscopic/ultrasonographic, are most significant and of great significance for the safe and successful performance of endoluminal vascular techniques. On balance, it is felt strongly that endoluminal catheter techniques are crucially important for the vascular surgeon of today and tomorrow. PMID- 8707803 TI - Principles of angioplasty guidance using ultrasound. AB - Marker bands that cast dense shadow on X-ray are placed at crucial points on a catheter (for example on a peripheral angioplasty catheter on either side of the balloon) so that those parts of the catheter can be guided inside the body by X ray. For catheter guidance by ultrasound, an ultrasound sensor in the mid-balloon region of an angioplasty catheter is integrated to the scanhead of an ultrasound scanner, via a catheter system interface (CSI). When the sensor in the catheter is imaged by the ultrasound scanner, the CSI injects a bright arrow corresponding to its position. This allows accurate catheter guidance using ultrasound. This paper describes the principles of ultrasound guidance of catheters and the initial clinical experience using this new catheter system (Echomark) in carrying out peripheral transluminal angioplasty. PMID- 8707804 TI - Challenges facing European vascular surgeons. PMID- 8707805 TI - Results of transluminal angioplasty of infra-popliteal arteries. AB - Transluminal angioplasty is rarely used to treat arterial diseases at leg level. During the last 5 years we have performed 25 distal artery angioplasties in 24 patients. Average age was 70 years, half of them were diabetics. Twenty patients had critical ischemia and 4 asymptomatic patients had critical stenosis distal to a previous patent infra-genicular bypass. Arteriography showed a single leg artery stenosis in 23 cases and a two distal artery stenosis in one case. No material other than guide wire was used to pass through the stenosis. All the dilation was performed in the operating room. Immediate results were good in 20 patients (21 stenosis). Four early failures occurred: one technical, one for progressive limb gangrene with secondary amputation, one postoperative thrombosis and one residual stenosis over 50%. Six patients had recurrent stenosis at mean 9 months. Five of them were treated with a new balloon angioplasty and the last with femoro-distal bypass. The 2 years cumulative primary and secondary patency rates were respectively 46 and 64%. With only guide wire and balloon catheters it is possible to treat distal artery lesions with rather good results. PMID- 8707806 TI - Endovascular remote endarterectomy in femoropopliteal long segmental occlusive disease. A new surgical technique illustrated and preliminary results using a ring strip cutter device. PMID- 8707807 TI - Stent graft for popliteal aneurysms. Six cases with Cragg Endo-Pro System 1 Mintec. AB - Six popliteal aneurysms were treated by transluminal exclusion with the Cragg Endopro System 1, a Stent-Graft manufactured by MINTEC. In one case, incomplete distal exclusion has led to a recurrent aneurysm at 6 months. In one case thrombosis occurred at 7 months, with successful reopening by fibrinolysis and an additional stent. The 4 other cases remained patent at 9, 8, 4 and 1 month. Despite these encouraging results, the stent-grafting of popliteal aneurysms seems still to be reserved for elderly or poor condition patient. PMID- 8707808 TI - Endovascular repair of iliac artery aneurysm with Endoprosystem I: a multicentric French study. AB - Due to their position deep in the pelvis, the classical surgical treatment of iliac artery aneurysm leads to a high morbidity and mortality rate. The transfemoral percutaneous repair of these aneurysms is now possible thanks to a new device the endoprosystem I from "Mintech". We began in 1994 a study including radiological, cardiac and vascular centers all of them skilled in endovascular procedures. 27 patients entered the study: 1 patient died, 2 attempt failed and 2 presented leakage: the immediate failure rate was then 18.5%. For the late result (min 6, Max 19, mean 12 months) we had 1 thrombosis treated by surgery, 1 restenosis treated by PTA. We did not see any polyester dilatation or reactivation of aneurysm at the scan control at one year. We conclude that the percutaneous treatment of iliac aneurysm is possible and safe but we need long term result to validate the method. PMID- 8707809 TI - The ideal guidance imaging system for endovascular interventions. AB - Endovascular interventions enable a minimally invasive approach to a variety of cardiovascular lesions. Small catheter-based delivery systems have rapidly evolved, diversifying endovascular therapeutic options. Coronary and peripheral vascular occlusive lesions may be amenable to angioplasty dilatation, intravascular stenting and ablation with mechanical atherectomy devices or lasers. New experimental transluminally placed endovascular stented grafts (TPEGs) have demonstrated feasibility for treating aneurysmal arteries, arteriovenous fistulas, and atherosclerotic occlusions. Improvements in imaging systems have provided better resolution of lesions, have improved patient selection and interventional strategy, and are invaluable for precise performance of procedures. The ideal guidance system should provide realtime imaging by a combination of complimentary modalities. These include cinefluoroscopy and contrast angiography, catheter-based intravascular ultrasound and angioscopy. PMID- 8707810 TI - Endovascular recanalization of the aorto-iliac segment: imaging and approaches. AB - Endovascular catheter techniques are most useful and successful in the aorto iliac arteries. As in other territories, wire recanalization, balloon angioplasty, and endoluminal stent placement are most applicable. For the surgeon, imaging considerations are most important since the operating room must be equipped with the proper capabilities for endovascular surgery. Techniques of catheterization and angiographic visualization of the arteries must be learned and understood clearly before these procedures are undertaken. A variety of occlusive lesions involving the aorta and iliac arteries can be treated adequately by means of nonsurgical catheter-based methods which add a significant new dimension to the vascular surgeon. PMID- 8707811 TI - Angioplasty of distal venous bypasses: is it worth the cost? AB - Saphenous vein is the best graft for long term patency infragenicular bypass. During follow-up, stenoses can appear on the graft. Is angioplasty a good solution for the treatment of these lesions? During the follow-up of 612 saphenous bypass with below-knee distal anastomosis, 90 stenoses (over 70%) were discovered. Among them 36 were treated with transluminal angioplasty (34 bypasses). Stenoses were detected at a mean follow-up of 7 months after bypass realization. They were located 17 times on the graft itself and 19 times near the anastomoses and they were never longer than 5 cm. Percutaneous approach was preferred for 11 cases and surgical for the others. Immediate success was obtained in 33 cases (91%). Among the 3 failed cases 2 needed a new bypass. Mean follow-up was 33 months after the initial bypass and 24 months after angioplasty. Graft patency was ensured by a single angioplasty in 18 cases. Assisted primary patency, cumulative patency, limb salvage rate were respectively 65%, 91%, 100% at one year and 53%, 72%, 96% at 2 years. Transluminal angioplasty can be advised for the treatment of short stenosis of infrainguinal vein graft: this technique has a weak risk, little surgical aggressiveness, short hospitalisation. Results are acceptable and not very different with the location of the stenosis. PMID- 8707812 TI - Long-term results with a Palmaz stent in the femoropopliteal arteries. AB - Highly contradictory results are achieved with stents in the superficial femoral and popliteal arteries. In order to compare our experience, we have reviewed a series of patients in whom a Palmaz stent was implanted at the femoropopliteal level. From January, 1991, to December, 1994, 35 patients were treated for claudication (63%) or for critical ischemia (37%), using endoluminal angioplasty and the implantation of a Palmaz stent. The stents were impianted in the superficial femoral artery in 26 cases (74%) and in the popliteal artery in 9 cases (26%). In case of restenosis (1 case), calcified stenosis (2 cases), complications of endoluminal angioplasty (32 cases), 44 stents were implanted in 35 stenoses (71.5%) and 10 short thromboses (28.5%). Follow-up was ensured periodically in all cases (D1, 2 months, 6 months, then every six months) using a clinical examination and a study with Doppler and duplex scanning. All stents were inserted without complications, with an initial success rate of 100%. During the follow-up period (32 +/- 4 months), two patients died (patent stent) and two were lost to follow-up. Acute thrombosis occurred in two cases (6%) and restenosis in five (14%) during the first six months after the operation. Restenosis occurred in all but one cases with a poor distal arterial runoff (p < 0.05). The popliteal artery was not more often affected by restenosis than the superficial femoral artery. Primary patency after one year is 80.4 +/- 7%, and 75.7 +/- 8% after two years. Assisted primary patency after 2 years is 83.3 +/- 7%. The implantation of a Palmaz stent is still a safe method, with low rates of acute thrombosis and a satisfactory long-term patency. Periodic and sonographic follow-up should allow improving the patency of restenosis by more than 50% under treatment. PMID- 8707813 TI - Cervical carotid artery stenosis: which technique, balloon angioplasty or surgery? AB - METHODS: Between April 1991 and November 1995, 38 patients mean age 65 (6 females, 32 males) were treated by cervical puncture for isolated cervical carotid stenosis (33 internal, 1 external, and 6 common carotid). All patients but 5 were symtomatic (19 TIA, 7 amarosis, 2 strokes and 4 VB symptoms). Complex lesions involving the carotid bifurcation and heavy calcifications were treated by conventional surgery. Two different groups of patients were considered. A first group of 19 patients (17 restenosis, 1 primary, 1 FM dysplasia) was treated by simple balloon angioplasty (BA). A second group of 19 patients was treated by primary stenting (16 DF NOVO, 2 radio-induced, and 2 recurrent stenosis). RESULTS: No hematoma required surgery. In the first group one patient died from an intracerebral hemorrhage, one presented a reversible stroke and 3 others a TIA. In the second group there was no complication or silent infarction on the CT SCAN: CONCLUSIONS: Balloon angioplasty appears to be associated with a high neurologic risk. While primary stenting seems much more reliable. A longer follow up to deal with restenosis. Conventional surgery remains the gold standard. PMID- 8707814 TI - Signal sequences specify the targeting route to the endoplasmic reticulum membrane. AB - In the yeast Saccharomyces cerevisiae, only a subset of preproteins that are translocated across the ER membrane require the function of the signal recognition particle (SRP), suggesting that an alternative, SRP-independent pathway must exist (Hann, B.C., and P. Walter. 1991. Cell. 67:131-144). We have established that the two targeting pathways function in parallel. Mutant alleles of SEC62 and SEC63 were isolated that specifically impaired the translocation of SRP-independent preproteins in vivo and in vitro, whereas SRP-dependent preproteins were unaffected. Based on this analysis, preproteins fall into three distinct classes: SRP dependent, SRP independent, and those that can use both pathways. Pathway specificity is conferred by the hydrophobic core of signal sequences. Our studies show a previously unrecognized diversity in ER-directed signal sequences, that carry structural information that serves to identify the route taken. PMID- 8707816 TI - Evidence that phospholipase D mediates ADP ribosylation factor-dependent formation of Golgi coated vesicles. AB - Formation of coatomer-coated vesicles from Golgi-enriched membranes requires the activation of a small GTP-binding protein, ADP ribosylation factor (ARF). ARF is also an efficacious activator of phospholipase D (PLD), an activity that is relatively abundant on Golgi-enriched membranes. It has been proposed that ARF, which is recruited onto membranes from cytosolic pools, acts directly to promote coatomer binding and is in a 3:1 stoichiometry with coatomer on coated vesicles. We present evidence that cytosolic ARF is not necessary for initiating coat assembly on Golgi membranes from cell lines with high constitutive PLD activity. Conditions are also described under which ARF is at most a minor component relative to coatomer in coated vesicles from all cell lines tested, including Chinese hamster ovary cells. Formation of coated vesicles was sensitive to ethanol at concentrations that inhibit the production of phosphatidic acid (PA) by PLD. When PA was produced in Golgi membranes by an exogenous bacterial PLD, rather than with ARF and endogenous PLD, coatomer bound to Golgi membranes. Purified coatomer also bound selectively to artificial lipid vesicles that contained PA and phosphatidylinositol (4,5)-bisphosphate (PIP2). We propose that activation of PLD and the subsequent production of PA are key early events for the formation of coatomer-coated vesicles. PMID- 8707815 TI - Endoplasmic reticulum localization of Sec12p is achieved by two mechanisms: Rer1p dependent retrieval that requires the transmembrane domain and Rer1p-independent retention that involves the cytoplasmic domain. AB - Yeast Sec12p is a type II transmembrane protein in the ER, which is essential for the formation of transport vesicles. From biochemical and morphological lines of evidence, we have proposed that Sec12p is localized to the ER by two mechanisms: static retention in the ER and dynamic retrieval from the early Golgi compartment. We have also shown that Rer1p, a membrane protein in the Golgi, is required for correct localization of Sec12p. In the present study, we have performed a systematic analysis to determine the ER localization signals in Sec12p corresponding to these two mechanisms. Both the transmembrane domain (TMD) and the NH2-terminal cytoplasmic domain of Sec12p show the ability to localize the protein to the ER. The effect of the TMD is potent and sufficient by itself for the ER localization and is strongly dependent on Rer1p. On the other hand, the cytoplasmic domain shows a moderate ER-localization capability which is independent of Rer1p. The rate of mannosyl modification has been measured to distinguish between retention and retrieval. The cytoplasmic domain significantly delays the transport from the ER to the cis-Golgi. In contrast, the TMD shows only a subtle retardation in the transport from the ER to the cis-Golgi but strictly prevents the transport beyond there. From these observations, we conclude that the TMD mainly acts as the retrieval signal and the cytoplasmic domain contains the retention signal. This study not only supports the two mechanisms hypothesis but also provides powerful tools to dissect the two. PMID- 8707817 TI - The human mitochondrial import receptor, hTom20p, prevents a cryptic matrix targeting sequence from gaining access to the protein translocation machinery. AB - Yeast Mas70p and NADH cytochrome b5 reductase are bitopic integral proteins of the mitochondrial outer membrane and are inserted into the lipid-bilayer in an Nin-Ccyto orientation via an NH2-terminal signal-anchor sequence. The signal anchor of both proteins is comprised of a short, positively charged domain followed by the predicted transmembrane segment. The positively charged domain is capable of functioning independently as a matrix-targeting signal in yeast mitochondria in vitro but does not support import into mammalian mitochondria (rat or human). Rather, this domain represents a cryptic signal that can direct import into mammalian mitochondria only if proximal components of the outer membrane import machinery are removed. This can be accomplished either by treating the surface of the intact mitochondria with trypsin or by generating mitoplasts. The import receptor Tom20p (Mas20p/MOM19) is responsible for excluding the cryptic matrix-targeting signal from mammalian mitochondria since replacement of yeast Tom20p with the human receptor confers this property to the yeast organelle while at the same time maintaining import of other proteins. In addition to contributing to positive recognition of precursor proteins, therefore, the results suggest that hTom20p may also have the ability to screen potential matrix-targeting sequences and exclude certain proteins that would otherwise be recognized and imported by distal components of the outer and inner membrane protein-translocation machinery. These findings also indicate, however, that cryptic signals, if they exist within otherwise native precursor proteins, may remain topogenically silent until the precursor successfully clears hTom20p, at which time the activity of the cryptic signal is manifested and can contribute to subsequent translocation and sorting of the polypeptide. PMID- 8707818 TI - Two components of the chloroplast protein import apparatus, IAP86 and IAP75, interact with the transit sequence during the recognition and translocation of precursor proteins at the outer envelope. AB - The interactions of precursor proteins with components of the chloroplast envelope were investigated during the early stages of protein import using a chemical cross-linking strategy. In the absence of energy, two components of the outer envelope import machinery, IAP86 and IAP75, cross-linked to the transit sequence of the precursor to the small subunit of ribulose-1, 5-bisphosphate carboxylase (pS) in a precursor binding assay. In the presence of concentrations of ATP or GTP that support maximal precursor binding to the envelope, cross linking to the transit sequence occurred predominantly with IAP75 and a previously unidentified 21-kD polypeptide of the inner membrane, indicating that the transit sequence had inserted across the outer membrane. Cross-linking of envelope components to sequences in the mature portion of a second precursor, preferredoxin, was detected in the presence of ATP or GTP, suggesting that sequences distant from the transit sequence were brought into the vicinity of the outer membrane under these conditions. IAP75 and a third import component, IAP34, were coimmunoprecipitated with IAP86 antibodies from solubilized envelope membranes, indicating that these three proteins form a stable complex in the outer membrane. On the basis of these observations, we propose that IAP86 and IAP75 act as components of a multisubunit complex to mediate energy-independent recognition of the transit sequence and subsequent nucleoside triphosphate induced insertion of the transit sequence across the outer membrane. PMID- 8707819 TI - Poisson-distributed active fusion complexes underlie the control of the rate and extent of exocytosis by calcium. AB - We have investigated the consequences of having multiple fusion complexes on exocytotic granules, and have identified a new principle for interpreting the calcium dependence of calcium-triggered exocytosis. Strikingly different physiological responses to calcium are expected when active fusion complexes are distributed between granules in a deterministic or probabilistic manner. We have modeled these differences, and compared them with the calcium dependence of sea urchin egg cortical granule exocytosis. From the calcium dependence of cortical granule exocytosis, and from the exposure time and concentration dependence of N ethylmaleimide inhibition, we determined that cortical granules do have spare active fusion complexes that are randomly distributed as a Poisson process among the population of granules. At high calcium concentrations, docking sites have on average nine active fusion complexes. PMID- 8707820 TI - Endocytosis of chimeric influenza virus hemagglutinin proteins that lack a cytoplasmic recognition feature for coated pits. AB - The influenza virus A/Japan/305/57 hemagglutinin (HA) can be converted from a protein that is essentially excluded from coated pits into one that is internalized at approximately the rate of uptake of bulk membrane by replacing the HA transmembrane and cytoplasmic sequences with those of either of two other glycoproteins (Roth et al., 1986. J. Cell Biol. 102:1271-1283). To identify more precisely the foreign amino acid sequences responsible for this change in HA traffic, DNA sequences encoding the transmembrane (TM) or cytoplasmic (CD) domains of either the G glycoprotein of vesicular stomatitis virus (VSV) or the gC glycoprotein of herpes simplex virus were exchanged for those encoding the analogous regions of wild type HA (HA wt). HA-HA-G and HA-HA-gC, chimeras that contain only a foreign CD, resembled HA wt in having a long residence on the cell surface and were internalized very slowly. HA-HA-gC was indistinguishable from HA in our assays, whereas twice as much HA-HA-G was internalized as was HA wt. However, HA-G-HA, containing only a foreign TM, was internalized as efficiently as was HA-G-G, a chimeric protein with transmembrane and cytoplasmic sequences of VSV G protein. Conditions that blocked internalization through coated pits also inhibited endocytosis of the chimeric proteins. Although the external domains of the chimeras were less well folded than that of the wild type HA, denaturation of the wild type HA external domain by treatment with low pH did not increase the interaction of HA with coated pits. However, mutation of four amino acids in the TM of HA allowed the protein to be internalized, indicating that the property that allows HA to escape endocytosis resides in its TM. These results indicate that possession of a cytoplasmic recognition feature is not required for the internalization of all cell surface proteins and suggest that multiple mechanisms for internalization exist that operate at distinctly different rates. PMID- 8707821 TI - Host cell invasion by trypanosomes requires lysosomes and microtubule/kinesin mediated transport. AB - Invasion of mammalian cells by the protozoan parasite Trypanosoma cruzi occurs by an actin-independent mechanism distinct from phagocytosis. Clusters of host lysosomes are observed at the site of parasite attachment, and lysosomal markers are detected in the vacuolar membrane at early stages of the entry process. These observations led to the hypothesis that the trypanosomes recruit host lysosomes to their attachment site, and that lysosomal fusion serves as a source of membrane to form the parasitophorous vacuole. Here we directly demonstrate directional migration of lysosomes to the parasite entry site, using time-lapse video-enhanced microscopy of L6E9 myoblasts exposed to T. cruzi trypomastigotes. BSA-gold-loaded lysosomes moved towards the cell periphery, in the direction of the parasite attachment site, but only when their original position was less than 11-12 microns from the invasion site. Lysosomes more distant from the invasion area exhibited only the short multi-directional saltatory movements previously described for lysosomes, regardless of their proximity to the cell margins. Specific depletion of peripheral lysosomes was obtained by microinjection of NRK cells with antibodies against the cytoplasmic domain of lgp 120, a treatment that aggregated lysosomes in the perinuclear area and inhibited T. cruzi entry. The microtubule-binding drugs nocodazole, colchicine, vinblastine, and taxol also inhibited invasion, in both NRK and L6E9 cells. Furthermore, microinjection of antibodies to the heavy chain of kinesin blocked the acidification-induced, microtubule-dependent redistribution of lysosomes to the host cell periphery, and reduced trypomastigote entry. Our results therefore demonstrate that during T. cruzi invasion of host cells lysosomes are mobilized from the immediately surrounding area, and that availability of lysosomes at the cell periphery and microtubule/kinesin-mediated transport are requirements for parasite entry. PMID- 8707822 TI - Sphingolipids and glycoproteins are differentially trafficked to the Chlamydia trachomatis inclusion. AB - Chlamydia trachomatis is an obligate intracellular pathogen that multiples within the confines of a membrane-bound vacuole called an inclusion. Approximately 40 50% of the sphingomyelin synthesized from exogenously added NBD-ceramide is specifically transported from the Golgi apparatus to the chlamydial inclusion (Hackstadt, T., M.A. Scidmore, and D.D. Rockey. 1995. Proc. Natl. Acad. Sci. USA. 92: 4877-4881). Given this major disruption of a cellular exocytic pathway and the similarities between glycolipid and glycoprotein exocytosis, we wished to determine whether the processing and trafficking of glycoproteins through the Golgi apparatus to the plasma membrane in chlamydia-infected cells was also disrupted. We analyzed the processing of several model glycoproteins including vesicular stomatitis virus G-protein, transferrin receptor, and human histocompatibility leukocyte class I antigen. In infected cells, the posttranslational processing and trafficking of these specific proteins through the Golgi apparatus and subsequent transport to the plasma membrane was not significantly impaired, nor were these glycoproteins found associated with the chlamydial inclusion membrane. Studies of receptor recycling from endocytic vesicles employing fluorescently and HRP-tagged transferrin and anti-transferrin receptor antibody revealed an increased local concentration of transferrin and transferrin receptor around but never within the chlamydial inclusion. However, Scatchard analysis failed to show either an increased intracellular accumulation of transferrin receptor or a decreased number of plasma membrane receptors in infected cells. Furthermore, the rate of exocytosis from the recycling endosomes to the plasma membrane was not altered in chlamydia-infected cells. Thus, although C. trachomatis disrupts the exocytosis of sphingolipids and the Golgi apparatus appears physically distorted, glycosylation and exocytosis of representative secreted and endocytosed proteins are not disrupted. These results suggest the existence of a previously unrecognized sorting of sphingolipids and glycoproteins in C. trachomatis-infected cells. PMID- 8707823 TI - Formation of triads without the dihydropyridine receptor alpha subunits in cell lines from dysgenic skeletal muscle. AB - Muscular dysgenesis (mdg/mdg), a mutation of the skeletal muscle dihydropyridine receptor (DHPR) alpha 1 subunit, has served as a model to study the functions of the DHPR in excitation-contraction coupling and its role in triad formation. We have investigated the question of whether the lack of the DHPR in dysgenic skeletal muscle results in a failure of triad formation, using cell lines (GLT and NLT) derived from dysgenic (mdg/mdg) and normal (+/+) muscle, respectively. The lines were generated by transfection of myoblasts with a plasmid encoding a Large T antigen. Both cell lines express muscle-specific proteins and begin organization of sarcomeres as demonstrated by immunocytochemistry. Similar to primary cultures, dysgenic (GLT) myoblasts show a higher incidence of cell fusion than their normal counterparts (NLT). NLT myotubes develop spontaneous contractile activity, and fluorescent Ca2+ recordings show Ca2+ release in response to depolarization. In contrast, GLTs show neither spontaneous nor depolarization-induced Ca2+ transients, but do release Ca2+ from the sarcoplasmic reticulum (SR) in response to caffeine. Despite normal transverse tubule (T tubule) formation, GLT myotubes lack the alpha 1 subunit of the skeletal muscle DHPR, and the alpha 2 subunit is mistargeted. Nevertheless, the ryanodine receptor (RyR) frequently develops its normal, clustered organization in the absence of both DHPR alpha subunits in the T-tubules. In EM, these RyR clusters correspond to T-tubule/SR junctions with regularly spaced feet. These findings provide conclusive evidence that interactions between the DHPR and RyR are not involved in the formation of triad junctions or in the normal organization of the RyR in the junctional SR. PMID- 8707824 TI - Coordinated regulation of platelet actin filament barbed ends by gelsolin and capping protein. AB - Exposure of cryptic actin filament fast growing ends (barbed ends) initiates actin polymerization in stimulated human and mouse platelets. Gelsolin amplifies platelet actin assembly by severing F-actin and increasing the number of barbed ends. Actin filaments in stimulated platelets from transgenic gelsolin-null mice elongate their actin without severing. F-actin barbed end capping activity persists in human platelet extracts, depleted of gelsolin, and the heterodimeric capping protein (CP) accounts for this residual activity. 35% of the approximately 5 microM CP is associated with the insoluble actin cytoskeleton of the resting platelet. Since resting platelets have an F-actin barbed end concentration of approximately 0.5 microM, sufficient CP is bound to cap these ends. CP is released from OG-permeabilized platelets by treatment with phosphatidylinositol 4,5-bisphosphate or through activation of the thrombin receptor. However, the fraction of CP bound to the actin cytoskeleton of thrombin stimulated mouse and human platelets increases rapidly to approximately 60% within 30 s. In resting platelets from transgenic mice lacking gelsolin, which have 33% more F-actin than gelsolin-positive cells, there is a corresponding increase in the amount of CP associated with the resting cytoskeleton but no change with stimulation. These findings demonstrate an interaction between the two major F-actin barbed end capping proteins of the platelet: gelsolin-dependent severing produces barbed ends that are capped by CP. Phosphatidylinositol 4,5 bisphosphate release of gelsolin and CP from platelet cytoskeleton provides a mechanism for mediating barbed end exposure. After actin assembly, CP reassociates with the new actin cytoskeleton. PMID- 8707825 TI - Smoothelin, a novel cytoskeletal protein specific for smooth muscle cells. AB - The characterization of a novel 59-kD cytoskeletal protein is described. It is exclusively observed in smooth muscle cells by Northern blotting and immunohistochemical analysis and therefore designated "smoothelin." A human smooth muscle cDNA library was screened with the monoclonal antibody R4A, and a full-size cDNA of the protein was selected. The cDNA was sequenced and appeared to contain a 1,113-bp open reading frame. Based on the cDNA sequence, the calculated molecular weight of the polypeptide was 40 kD and it was demonstrated to contain two N-glycosylation sites. Computer assisted analysis at the protein level revealed a 56-amino acid domain with homologies of approximately 40% with a sequence bordering the actin-binding domains of dystrophin, utrophin, beta spectrin and alpha-actinin. In situ hybridization demonstrated that human smoothelin is encoded by a single copy gene which is located on chromosome 22. Immunohistochemistry and Western blotting revealed synthesis of smoothelin in smooth muscle of species evolutionarily as far apart as human and teleost. Northern blotting indicated that sequence as well as size of the mRNA (approximately 1,500 bases) are conserved among vertebrates. Cell fractionation studies and differential centrifugation showed that the protein cannot be extracted with Triton X-100, which indicates that it is a part of the cytoskeleton. Transfection of the human cDNA into smooth muscle cells and COS7 cells produced a protein of 59 kD, which assembled into a filamentous network. However, in rat heart-derived myoblasts association with stress fibers was most prominent. Smoothelin was not detected in primary or long term smooth muscle cell cultures. Also, transcription of smoothelin mRNA was almost instantly halted in smooth muscle tissue explants. We conclude that smoothelin is a new cytoskeletal protein that is only found in contractile smooth muscle cells and does not belong to one of the classes of structural proteins presently known. PMID- 8707826 TI - The BUD4 protein of yeast, required for axial budding, is localized to the mother/BUD neck in a cell cycle-dependent manner. AB - A and alpha cells of the yeast Saccharomyces cerevisiae exhibit an axial budding pattern, whereas a/alpha diploid cells exhibit a bipolar pattern. Mutations in BUD3, BUD4, and AXL1 cause a and alpha cells to exhibit the bipolar pattern, indicating that these genes are necessary to specify the axial budding pattern (Chant, J., and I. Herskowitz. 1991. Cell. 65:1203-1212; Fujita, A., C. Oka, Y. Arikawa, T. Katagi, A. Tonouchi, S. Kuhara, and Y. Misumi. 1994. Nature (Lond.). 372:567-570). We cloned and sequenced BUD4, which codes for a large, novel protein (Bud4p) with a potential GTP-binding motif. Bud4p is expressed and localized to the mother/bud neck in all cell types. Most mitotic cells contain two apparent rings of Bud4 immunoreactive staining, as observed for Bud3p (Chant, J., M. Mischke, E. Mitchell, I. Herskowitz, and J.R. Pringle. 1995. J. Cell Biol. 129: 767-778). Early G1 cells contain a single ring of Bud4p immunoreactive staining, whereas cells at START and in S phase lack these rings. The level of Bud4p is also regulated in a cell cycle-dependent manner. Bud4p is inefficiently localized in bud3 mutants and after a temperature shift of a temperature sensitive mutant, cdc12, defective in the neck filaments. These observations suggest that Bud4p and Bud3p cooperate to recognize a spatial landmark (the neck filaments) during mitosis and support the hypothesis that they subsequently become a landmark for establishing the axial budding pattern in G1. PMID- 8707827 TI - gamma-Tubulin-like Tub4p of Saccharomyces cerevisiae is associated with the spindle pole body substructures that organize microtubules and is required for mitotic spindle formation. AB - Tub4p is a novel tubulin in Saccharomyces cerevisiae that most closely resembles gamma-tubulin. We report in this manuscript that the essential Tub4p is associated with the inner and outer plaques of the yeast microtubule organizing center, the spindle pole body (SPB). These SPB substructures are involved in the attachment of the nuclear and cytoplasmic microtubules, respectively (Byers, B., and L. Goetsch. 1975. J. Bacteriol. 124:511-523). Study of a temperature sensitive tub4-1 allele revealed that TUB4 has essential functions in microtubule organization. Remarkably, SPB duplication and separation are not impaired in tub4 1 cells incubated at the nonpermissive temperature. However, SPBs from such cells contain less or misdirected nuclear microtubules. Further analysis revealed that tub4-1 cells are able to assemble a short bipolar spindle, suggesting that the defect in microtubule organization occurs after spindle formation. A role of Tub4p in microtubule organization is further suggested by an increase in chromosome loss in tub4-1 cells. In addition, cell cycle arrest and survival of tub4-1 cells is dependent on the mitotic checkpoint control gene BUB2 (Hoyt, M.A., L. Totis, B.T. Roberts. 1991. Cell. 66:507-517), one of the cell's monitors of spindle integrity. PMID- 8707828 TI - Analysis of Tub4p, a yeast gamma-tubulin-like protein: implications for microtubule-organizing center function. AB - gamma-Tubulin is a conserved component of microtubule-organizing centers and is thought to be involved in microtubule nucleation. A recently discovered Saccharomyces cerevisiae gene (TUB4) encodes a tubulin that is related to, but divergent from, gamma-tubulins. TUB4 is essential for cell viability, and epitope tagged Tub4 protein (Tub4p) is localized to the spindle pole body (Sobel, S.G., and M. Snyder. 1995.J. Cell Biol. 131:1775-1788). We have characterized the expression of TUB4, the association of Tub4p with the spindle pole body, and its role in microtubule organization. Tub4p is a minor protein in the cell, and expression of TUB4 is regulated in a cell cycle-dependent manner. Wild-type Tub4p is localized to the spindle pole body, and a Tub4p-green fluorescent protein fusion is able to associate with a preexisting spindle pole body, suggesting that there is dynamic exchange between cytoplasmic and spindle pole body forms of Tub4p. Perturbation of Tub4p function, either by conditional mutation or by depletion of the protein, results in spindle as well as spindle pole body defects, but does not eliminate the ability of microtubules to regrow from, or remain attached to, the spindle pole body. The spindle pole bodies in tub4 mutant cells duplicate but do not separate, resulting in a monopolar spindle. EM revealed that one spindle pole body of the duplicated pair appears to be defective for the nucleation of microtubules. These results offer insight into the role of gamma-tubulin in microtubule-organizing center function. PMID- 8707829 TI - Anastral meiotic spindle morphogenesis: role of the non-claret disjunctional kinesin-like protein. AB - We have used time-lapse laser scanning confocal microscopy to directly examine microtubule reorganization during meiotic spindle assembly in living Drosophila oocytes. These studies indicate that the bipolarity of the meiosis I spindle is not the result of a duplication and separation of centrosomal microtubule organizing centers (MTOCs). Instead, microtubules first associate with a tight chromatin mass, and then bundle to form a bipolar spindle that lacks asters. Analysis of mutant oocytes indicates that the Non-Claret Disjunctional (NCD) kinesin-like protein is required for normal spindle assembly kinetics and stabilization of the spindle during metaphase arrest. Immunolocalization analyses demonstrate that NCD is associated with spindle microtubules, and that the centrosomal components gamma-tubulin, CP-190, and CP-60 are not concentrated at the meiotic spindle poles. Based on these observations, we propose that microtubule bundling by the NCD kinesin-like protein promotes assembly of a stable bipolar spindle in the absence of typical MTOCs. PMID- 8707831 TI - Weaver granule neurons are rescued by calcium channel antagonists and antibodies against a neurite outgrowth domain of the B2 chain of laminin. AB - The weaver mutation impairs migration of the cerebellar granular neurons and induces neuronal death during the first two weeks of postnatal life. To elucidate the molecular mechanisms for the impaired neuronal migration, we investigated the rescue mechanisms of the weaver (wv/wv) granule neurons in vitro. We found that Fab2 fragments of antibodies against a neurite outgrowth domain of the B2 chain of laminin enhanced neurite outgrowth and neuronal migration of the weaver granule neurons on a laminin substratum and in the established cable culture system. The rescue of the weaver granule neurons by antibodies against the B2 chain of laminin may result from the neutralizing effect of these antibodies against the elevated B2 chain levels of the weaver brain. The L-type calcium channel blocker, verapamil (1-5 microM), also rescued the weaver granule neurons. High concentrations of MK-801 (10-20 microM), a glutamate receptor antagonist and voltage-gated calcium channel blocker, rescued the weaver granule neurons similar to verapamil, but low concentrations of MK-801 (1 microM) had no rescue effect. Simultaneous patch-clamp studies indicated that the weaver granule neurons did not express functional N-methyl-D-aspartate receptors further indicating that the rescue of the weaver granule neurons by MK-801 resulted from its known inhibition of voltage-gated calcium channels. The present results indicate that antibodies against the B2 chain of laminin, verapamil, and high concentrations of MK-801 protect the weaver granule neurons from the otherwise destructive action of the weaver gene. Thus, both the laminin system and calcium channel function contribute to the migration deficiency of the weaver granule neurons. PMID- 8707830 TI - ARIA/HRG regulates AChR epsilon subunit gene expression at the neuromuscular synapse via activation of phosphatidylinositol 3-kinase and Ras/MAPK pathway. AB - AChR-inducing activity (ARIA)/heregulin, a ligand for erbB receptor tyrosine kinases (RTKs), is likely to be one nerve-supplied signal that induces expression of acetylcholine receptor (AChR) genes at the developing neuromuscular junction. Since some RTKs act through Ras and phosphatidylinositol 3-kinase (PI3K), we investigated the role of these pathways in ARIA signaling. Expression of activated Ras or Raf mimicked ARIA-induction of AChR epsilon subunit genes in muscle cells; whereas dominant negative Ras or Raf blocked the effect of ARIA. ARIA rapidly activated erk1 and erk2 and inhibition of both erks also abolished the effect of ARIA. ARIA stimulated association of PI3K with erbB3, expression of an activated PI3K led to ARIA-independent AChR epsilon subunit expression, and inhibition of PI3K abolished the action of ARIA. Thus, synaptic induction of AChR genes requires activation of both Ras/MAPK and PI3K signal transduction pathways. PMID- 8707832 TI - Cutaneous overexpression of NT-3 increases sensory and sympathetic neuron number and enhances touch dome and hair follicle innervation. AB - Target-derived influences of nerve growth factor on neuronal survival and differentiation are well documented, though effects of other neurotrophins are less clear. To examine the influence of NT-3 neurotrophin overexpression in a target tissue of sensory and sympathetic neurons, transgenic mice were isolated that overexpress NT-3 in the epidermis. Overexpression of NT-3 led to a 42% increase in the number of dorsal root ganglia sensory neurons, a 70% increase in the number of trigeminal sensory neurons, and a 32% increase in sympathetic neurons. Elevated NT-3 also caused enlargement of touch dome mechanoreceptor units, sensory end organs innervated by slowly adapting type 1 (SA1) neurons. The enlarged touch dome units of the transgenics had an increased number of associated Merkel cells, cells at which SA1s terminate. An additional alteration of skin innervation in NT-3 transgenics was an increased density of myelinated circular endings associated with the piloneural complex. The enhancement of innervation to the skin was accompanied by a doubling in the number of sensory neurons expressing trkC. In addition, measures of nerve fibers in cross-sectional profiles of cutaneous saphenous nerves of transgenics showed a 60% increase in myelinated fibers. These results indicate that in vivo overexpression of NT-3 by the epidermis enhances the number of sensory and sympathetic neurons and the development of selected sensory endings of the skin. PMID- 8707833 TI - Activity-induced internalization and rapid degradation of sodium channels in cultured fetal neurons. AB - A regulatory mechanism for neuronal excitability consists in controlling sodium channel density at the plasma membrane. In cultured fetal neurons, activation of sodium channels by neurotoxins, e.g., veratridine and alpha-scorpion toxin (alpha ScTx) that enhance the channel open state probability induced a rapid down regulation of surface channels. Evidence that the initial step of activity induced sodium channel down-regulation is mediated by internalization was provided by using 125I-alpha-ScTx as both a channel probe and activator. After its binding to surface channels, the distribution of 125I-alpha-ScTx into five subcellular compartments was quantitatively analyzed by EM autoradiography. 125I alpha-ScTx was found to accumulate in tubulovesicular endosomes and disappear from the cell surface in a time-dependent manner. This specific distribution was prevented by addition of tetrodotoxin (TTX), a channel blocker. By using a photoreactive derivative to covalently label sodium channels at the surface of cultured neurons, we further demonstrated that they are degraded after veratridine-induced internalization. A time-dependent decrease in the amount of labeled sodium channel alpha subunit was observed after veratridine treatment. After 120 min of incubation, half of the alpha subunits were cleaved. This degradation was prevented totally by TTX addition and was accompanied by the appearance of an increasing amount of a 90-kD major proteolytic fragment that was already detected after 45-60 min of veratridine treatment. Exposure of the photoaffinity-labeled cells to amphotericin B, a sodium ionophore, gave similar results. In this case, degradation was prevented when Na+ ions were substituted by choline ions and not blocked by TTX. After veratridine- or amphotericin B induced internalization of sodium channels, breakdown of the labeled alpha subunit was inhibited by leupeptin, while internalization was almost unaffected. Thus, cultured fetal neurons are capable of adjusting sodium channel density by an activity-dependent endocytotic process that is triggered by Na+ influx. PMID- 8707835 TI - Nuclear Translocation of fibroblast growth factor (FGF) receptors in response to FGF-2. AB - Members of the FGF family of growth factors localize to the nuclei in a variety of different cell types. To determine whether FGF receptors are also present within nuclei and if this localization is regulated by FGFs, nuclei were prepared from quiescent and FGF-2-treated Swiss 3T3 fibroblasts and examined for the presence of FGF receptors by immunoblotting with an antibody produced against the extracellular domain of FGF receptor-1 (FGFR-1). Little or no FGFR-1 is detected in nuclei prepared from quiescent cells. When cells are treated with FGF-2, however, there is a time- and dose-dependent increase in the association of FGFR 1 immunoreactivity with the nucleus. In contrast, treatment with either EGF or 10% serum does not increase the association of FGFR-1 with the nucleus. When cell surface proteins are labeled with biotin, a biotinylated FGFR-1 is detected in the nuclear fraction prepared from FGF-2-treated, but not untreated, cells indicating that the nuclear-associated FGFR-1 immunoreactivity derives from the cell surface. The presence of FGFR-1 in the nuclei of FGF-2-treated cells was confirmed by immunostaining with a panel of different FGFR-1 antibodies, including one directed against the COOH-terminal domain of the protein. Fractionation of nuclei from FGF-2-treated cells indicates that nuclear FGFR-1 is localized to the nuclear matrix, suggesting that the receptor may play a role in regulating gene activity. PMID- 8707834 TI - Biochemical properties and subcellular distribution of the BI and rbA isoforms of alpha 1A subunits of brain calcium channels. AB - Biochemical properties and subcellular distribution of the class A calcium channel alpha 1 subunits (alpha 1A) from rat and rabbit brain were examined using site-directed anti-peptide antibodies specific for rat rbA (anti-CNA3) and for rabbit BI (anti-NBI-1 and anti-NBI-2) isoforms of alpha 1A. In immunoblotting experiments, anti-CNA3 specifically identifies multiple alpha 1A polypeptides with apparent molecular masses of 210, 190, and 160 kD, and anti-NBI-1 and anti NBI-2 specifically recognize 190-kD alpha 1A polypeptides in rat brain membrane. In rabbit brain, anti-NBI-1 or anti-NBI-2 specifically detect alpha 1A polypeptides with apparent molecular masses of 220, 200, and 190 kD, while anti CNA3 specifically recognizes 190-kD alpha 1A polypeptides. These polypeptides evidently represent multiple isoforms of alpha 1A present in both rat and rabbit brain. Anti-CNA3 specifically immunoprecipitates high affinity receptor sites for omega-conotoxin MVIIC (Kd approximately 100 pM), whereas anti-NBI-2 immunoprecipitates two distinct affinity receptor sites for omega-conotoxin MVIIC (Kd approximately 100 pM and approximately 1 microM). Coimmunoprecipitation experiments indicate that alpha 1A subunits recognized by anti-CNA3 and anti-NBI 2 are associated with syntaxin in a stable, SDS-resistant complex and with synaptotagmin. Immunofluorescence studies reveal that calcium channels recognized by anti-NBI-2 are localized predominantly in dendrites and nerve terminals forming synapses on them, while calcium channels recognized by anti-CNA3 are localized more prominently in cell bodies and in nerve terminals. The mossy fiber terminals in hippocampus and the terminals of climbing and parallel fibers in cerebellum are differentially stained by these isoform-specific antibodies. These results indicate that both rbA and BI isoforms of alpha 1A are expressed in rat and rabbit brain and form calcium channels having alpha 1A subunits with distinct molecular mass, pharmacology, and subcellular localization. PMID- 8707836 TI - eat-5 and unc-7 represent a multigene family in Caenorhabditis elegans involved in cell-cell coupling. AB - The Drosophila melanogaster genes Passover and l(1)ogre and the Caenorhabditis elegans gene unc-7 define a gene family whose function is not known. We have isolated and characterized the C. elegans gene eat-5, which is required for synchronized pharyngeal muscle contractions, and find that it is a new member of this family. Simultaneous electrical and video recordings reveal that in eat-5 mutants, action potentials of muscles in the anterior and posterior pharynx are unsynchronized. Injection of carboxyfluorescein into muscles of the posterior pharynx demonstrates that all pharyngeal muscles are dye-coupled in wild-type animals; in eat-5 mutants, however, muscles of the anterior pharynx are no longer dye-coupled to posterior pharyngeal muscles. We show that a gene fusion of eat-5 to the green fluorescent protein is expressed in pharyngeal muscles. unc-7 and eat-5 are two of at least sixteen members of this family in C. elegans as determined by database searches and PCR-based screens. The amino acid sequences of five of these members in C. elegans have been deduced from cDNA sequences. Polypeptides of the family are predicted to have four transmembrane domains with cytoplasmic amino and carboxyl termini. We have constructed fusions of one of these polypeptides with beta-galactosidase and with green fluorescent protein. The fusion proteins appear to be localized in a punctate pattern at or near plasma membranes. We speculate that this gene family is required for the formation of gap junctions. PMID- 8707837 TI - Mechanism for transition from initial to stable cell-cell adhesion: kinetic analysis of E-cadherin-mediated adhesion using a quantitative adhesion assay. AB - A centrifugal force-based adhesion assay has been used to quantitatively examine the kinetics of formation of cell-cell contacts mediated specifically by expression of E-cadherin under the control of a glucocorticoid-inducible promoter in mouse fibroblasts. Analysis of cells expressing maximal or minimal levels of E cadherin showed that the strength of E-cadherin-mediated adhesion developed in a single exponential step over a short time (half-maximal adhesion, 13-17 min). At 37 degrees C, adhesion strength increased rapidly in the first 20 min without an apparent lag phase. After 90 min, adhesion strength reached a plateau. Differences in final strengths of adhesion were commensurate with the level of E cadherin expression. Strengthening of adhesion was temperature dependent. At 19 degrees C, strengthening of adhesion was delayed and subsequently developed with a slower rate compared to adhesion at 37 degrees C. At 4 degrees C, adhesion was completely inhibited. Strengthening of adhesion was absolutely dependent on a functional actin cytoskeleton since adhesion did not develop when cells were treated with cytochalasin D. Together, our current and previous (McNeill et al., 1993.J. Cell Biol. 120:1217-1226) studies indicate that the rate of initial strengthening of E-cadherin-mediated adhesion is neither dependent on the amount of E-cadherin expressed nor on long-range protein diffusion in the membrane to the adhesion site. However, initial strengthening of adhesion is dependent on temperature-sensitive cellular activities that may locally couple clusters of E cadherin to the actin cytoskeleton. PMID- 8707838 TI - Beta4 integrin is required for hemidesmosome formation, cell adhesion and cell survival. AB - The integrin heterodimer alpha 6 beta 4 is expressed in many epithelia and in Schwann cells. In stratified epithelia, alpha 6 beta 4 couple with BPAG1-e and BPAG2 to form hemidesmosomes, attaching externally to laminin and internally to the keratin cytoskeleton. To explore the function of this atypical integrin, and its relation to conventional actin-associated integrins, we targeted the removal of the beta 4 gene in mice. Tissues that express alpha 6 beta 4 are grossly affected. Stratified tissues are devoid of hemidesmosomes, display only a very fragile attachment to the basal lamina, and exhibit signs of degeneration and tissue disorganization. Simple epithelia which express alpha 6 beta 4 are also defective in adherence, even though they do not form hemidesmosomes. In the absence of beta 4, alpha 6 is dramatically downregulated, and other integrins do not appear to compensate for the loss of this heterodimer. These data have important implications for understanding integrin function in cell-substratum adhesion, cell survival and differentiation, and for understanding the role of alpha 6 beta 4 in junctional epidermolysis bullosa, an often lethal human disorder with pathology similar to our mice. PMID- 8707839 TI - Altered rate of fibronectin matrix assembly by deletion of the first type III repeats. AB - The assembly of fibronectin (FN) into a fibrillar matrix is a complex stepwise process that involves binding to integrin receptors as well as interactions between FN molecules. To follow the progression of matrix formation and determine the stages during which specific domains function, we have developed cell lines that lack an endogenous FN matrix but will form fibrils when provided with exogenous FN. Recombinant FNs (recFN) containing deletions of either the RGD cell binding sequence (RGD-) or the first type III repeats (FN delta III1-7) including the III1 FN binding site were generated with the baculovirus insect cell expression system. After addition to cells, recFN matrix assembly was monitored by indirect immunofluorescence and by insolubility in the detergent deoxycholate (DOC). In the absence of any native FN, FN delta III1-7 was assembled into fibrils and was converted into DOC-insoluble matrix. This process could be inhibited by the amino-terminal 70 kD fragment of FN, showing that FN delta III1 7 follows an assembly pathway similar to FN. The progression of FN delta III1-7 assembly differed from native FN in that the recFN became DOC-insoluble more quickly. In contrast, RGD- recFNs were not formed into fibrils except when added in combination with native FN. These results show that the RGD sequence is essential for the initiation step but fibrils can form independently of the III1 7 modules. The altered rate of FN delta III1-7 assembly suggests that one function of the missing repeats might be to modulate an early stage of matrix formation. PMID- 8707840 TI - Molecular and functional characterization of the p62 complex, an assembly of nuclear pore complex glycoproteins. AB - Macromolecular trafficking across the nuclear envelope involves interactions between cytosolic transport factors and nuclear pore complex proteins. The p62 complex, an assembly of 62, 58, 54, and 45-kD O-linked glycoproteins-localized near the central gated channel of the nuclear pore complex, has been directly implicated in nuclear protein import. The cDNA cloning of rat p62 was reported previously. We have now carried out cDNA cloning of rat p58, p54, and p45. We found that p58 contains regions with FG (Phe, Gly) and PA (Pro, Ala) repeats at both its NH2 and COOH termini separated by a predicted alpha-helical coiled-coil region, while p54 has an NH2-terminal FG and PA repeat region and a COOH-terminal predicted coiled-coil region. p45 and p58 appear to be generated by alternative splicing, with p45 containing the NH2-terminal FG repeat region and the coiled coil region of p58. Using immunogold electron microscopy, we found that p58/p45 and p54 are localized on both sides of the nuclear pore complex, like p62. Previous studies have shown that immobilized recombinant p62 can bind the cytosolic nuclear import factor NTF2 and thereby deplete transport activity from cytosol. We have now found that immobilized recombinant p58 and p54 also can deplete nuclear transport activity from cytosol, and that p62, p58, and p54 bind directly to the cytosolic nuclear import factors p97 and NTF2. At least in the case of p58, this involves FG repeat regions. Moreover, p58 can bind to a complex containing transport ligand, the nuclear localization sequence receptor (Srp1 alpha) and p97. These data support a model in which the p62 complex binds to a multicomponent particle consisting of transport ligand and cytosolic factors to achieve accumulation of ligand near the central gated channel of the nuclear pore complex. PMID- 8707842 TI - Neither type of mannose 6-phosphate receptor is sufficient for targeting of lysosomal enzymes along intracellular routes. AB - Mouse embryonic fibroblasts that are deficient in the two mannose 6-phosphate receptors (MPRs) MPR 46 and MPR 300 missort the majority (> or = 85%) of soluble lysosomal proteins into the medium. Human MPR 46 and MPR 300 were expressed in these cells to test whether overexpression of a single type of MPR can restore transport of lysosomal proteins to lysosomes. Only a partial correction of the missorting was observed after overexpression of MPR 46. Even at MPR 46 levels that are five times higher than the wild-type level, more than one third of the newly synthesized lysosomal proteins accumulates in the secretions. Two-fold overexpression of MPR 300 completely corrects the missorting of lysosomal enzymes. However, at least one fourth of the lysosomal enzymes are transported along a secretion-recapture pathway that is sensitive to mannose 6-phosphate in medium. In control fibroblasts that express both types of MPR, the secretion recapture pathway is of minor importance. These results imply that neither overexpression of MPR 46 nor MPR 300 is sufficient for targeting of lysosomal proteins along intracellular routes. PMID- 8707841 TI - The cold sensitivity of a mutant of Saccharomyces cerevisiae lacking a mitochondrial heat shock protein 70 is suppressed by loss of mitochondrial DNA. AB - SSH1, a newly identified member of the heat shock protein (hsp70) multigene family of the budding yeast Saccharomyces cerevisiae, encodes a protein localized to the mitochondrial matrix. Deletion of the SSH1 gene results in extremely slow growth at 23 degrees C or 30 degrees C, but nearly wild-type growth at 37 degrees C. The matrix of the mitochondria contains another hsp70, Ssc1, which is essential for growth and required for translocation of proteins into mitochondria. Unlike SSC1 mutants, an SSH1 mutant showed no detectable defects in import of several proteins from the cytosol to the matrix compared to wild type. Increased expression of Ssc1 partially suppressed the cold-sensitive growth defect of the SSH1 mutant, suggesting that when present in increased amounts, Ssc1 can at least partially carry out the normal functions of Ssh1. Spontaneous suppressors of the cold-sensitive phenotype of an SSH1 null mutant were obtained at a high frequency at 23 degrees C, and were all found to be respiration deficient. 15 of 16 suppressors that were analyzed lacked mitochondrial DNA, while the 16th had reduced amounts. We suggest that Ssh1 is required for normal mitochondrial DNA replication, and that disruption of this process in ssh1 cells results in a defect in mitochondrial function at low temperatures. PMID- 8707843 TI - The glucose transporter (GLUT-4) and vesicle-associated membrane protein-2 (VAMP 2) are segregated from recycling endosomes in insulin-sensitive cells. AB - Insulin stimulates glucose transport in adipocytes by translocation of the glucose transporter (GLUT-4) from an intracellular site to the cell surface. We have characterized different synaptobrevin/vesicle-associated membrane protein (VAMP) homologues in adipocytes and studied their intracellular distribution with respect to GLUT-4. VAMP-1, VAMP-2, and cellubrevin cDNAs were isolated from a 3T3 L1 adipocyte expression library. VAMP-2 and cellubrevin were: (a) the most abundant isoforms in adipocytes, (b) detectable in all insulin responsive tissues, (c) translocated to the cell surface in response to insulin, and (d) found in immunoadsorbed GLUT-4 vesicles. To further define their intracellular distribution, 3T3-L1 adipocytes were incubated with a transferrin/HRP conjugate (Tf/HRP) and endosomes ablated following addition of DAB and H2O2. While this resulted in ablation of > 90% of the transferrin receptor (TfR) and cellubrevin found in intracellular membranes, 60% of GLUT-4 and 90% of VAMP-2 was not ablated. Immuno-EM on intracellular vesicles from adipocytes revealed that VAMP-2 was colocalized with GLUT-4, whereas only partial colocalization was observed between GLUT-4 and cellubrevin. These studies show that two different v-SNAREs, cellubrevin and VAMP-2, are partially segregated in different intracellular compartments in adipocytes, implying that they may define separate classes of secretory vesicles in these cells. We conclude that a proportion of GLUT-4 is found in recycling endosomes in nonstimulated adipocytes together with cellubrevin and the transferrin receptor. In addition, GLUT-4 and VAMP-2 are selectively enriched in a postendocytic compartment. Further study is required to elucidate the function of this latter compartment in insulin-responsive cells. PMID- 8707844 TI - ZP3-dependent activation of sperm cation channels regulates acrosomal secretion during mammalian fertilization. AB - The sperm acrosome reaction is a Ca(2+)-dependent secretory event required for fertilization. Adhesion to the egg's zona pellucida promotes Ca2+ influx through voltage-sensitive channels, thereby initiating secretion. We used potentiometric fluorescent probes to determine the role of sperm membrane potential in regulating Ca2+ entry. ZP3, the glycoprotein agonist of the zona pellucida, depolarizes sperm membranes by activating a pertussis toxin-insensitive mechanism with the characteristics of a poorly selective cation channel. ZP3 also activates a pertussis toxin-sensitive pathway that produces a transient rise in internal pH. The concerted effects of depolarization and alkalinization open voltage sensitive Ca2+ channels. These observations suggest that mammalian sperm utilize membrane potential-dependent signal transduction mechanisms and that a depolarization pathway is an upstream transducing element coupling adhesion to secretion during fertilization. PMID- 8707845 TI - Lipid-modified, cysteinyl-containing peptides of diverse structures are efficiently S-acylated at the plasma membrane of mammalian cells. AB - A variety of cysteine-containing, lipid-modified peptides are found to be S acylated by cultured mammalian cells. The acylation reaction is highly specific for cysteinyl over serinyl residues and for lipid-modified peptides over hydrophilic peptides. The S-acylation process appears by various criteria to be enzymatic and resembles the S-acylation of plasma membrane-associated proteins in various characteristics, including inhibition by tunicamycin. The substrate range of the S-acylation reaction encompasses, but is not limited to, lipopeptides incorporating the motifs myristoylGC- and -CXC(farnesyl)-OCH3, which are reversibly S-acylated in various intracellular proteins. Mass-spectrometric analysis indicates that palmitoyl residues constitute the predominant but not the only type of S-acyl group coupled to a lipopeptide carrying the myristoylGC- motif, with smaller amounts of S-stearoyl and S-oleoyl substituents also detectable. Fluorescence microscopy using NBD-labeled cysteinyl lipopeptides reveals that the products of lipopeptide S-acylation, which cannot diffuse between membranes, are in almost all cases localized preferentially to the plasma membrane. This preferential localization is found even at reduced temperatures where vesicular transport from the Golgi complex to the plasma membrane is suppressed, strongly suggesting that the plasma membrane itself is the preferred site of S-acylation of these species. Uniquely among the lipopeptides studied, species incorporating an unphysiological N-myristoylcysteinyl- motif also show substantial formation of S-acylated products in a second, intracellular compartment identified as the Golgi complex by its labeling with a fluorescent ceramide. Our results suggest that distinct S-acyltransferases exist in the Golgi complex and plasma membrane compartments and that S-acylation of motifs such as myristoylGC- occurs specifically at the plasma membrane, affording efficient targeting of cellular proteins bearing such motifs to this membrane compartment. PMID- 8707846 TI - Ubiquitination of the yeast a-factor receptor. AB - The a-factor receptor (Ste3p) is one of two pheromone receptors in the yeast Saccharomyces cerevisiae that enable the cell-cell communication of mating. In this report, we show that this receptor is subject to two distinct covalent modifications-phosphorylation and ubiquitination. Phosphorylation, evident on the unstimulated receptor, increases upon challenge by the receptor's ligand, a factor. We suggest that this phosphorylation likely functions in the adaptive, negative regulation of receptor activity. Removal of phosphorylation by phosphatase treatment uncovered two phosphatase-resistant modifications identified as ubiquitination using a myc-epitope-tagged ubiquitin construct. Ste3p undergoes rapid, ligand-independent turnover that depends on vacuolar proteases and also on transport of the receptor from surface to vacuole (i.e., endocytosis) (Davis, N.G., J.L.Horecka, and G.F. Sprague, Jr., 1993 J. Cell Biol. 122:53-65). An end4 mutation, isolated for its defect in the endocytic uptake of alpha-factor pheromone (Raths, S., J. Rohrer, F. Crausaz, and H. Riezman. 1993. J. Cell Biol. 120:55-65), blocks constitutive endocytosis of the a-factor receptor, yet fails to block ubiquitination of the receptor. In fact, both phosphorylation and ubiquitination of the surfacebound receptor were found to increase, suggesting that these modifications may occur normally while the receptor is at the cell surface. In a mutant strain constructed to allow for depletion of ubiquitin, the level of receptor ubiquitination was found to be substantially decreased. Correlated with this was an impairment of receptor degradative turnover-receptor half-life that is normally approximately 20 min at 30 degrees C was increased to approximately 2 h under these ubiquitin-depletion conditions. Furthermore, surface residency, normally of short duration in wild type cells (terminated by endocytosis to the vacuole), was found to be prolonged; the majority of the receptor protein remained surface localized fully 2 h after biosynthesis. Thus, the rates of a-factor receptor endocytosis and consequent vacuolar turnover depend on the available level of ubiquitin in the cell. In cells mutant for two E2 activities, i.e., ubc4 delta ubc5 delta cells, the receptor was found to be substantially less ubiquitinated, and in addition, receptor turnover was slowed, suggesting that Ubc4p and Ubc5p may play a role in the recognition of the receptor protein as substrate for the ubiquitin system. In addition to ligand-independent uptake, the a-factor receptor also undergoes a ligand-dependent form of endocytosis (Davis, N.G., J.L. Horecka, and G.F. Sprague, Jr. 1993. J. Cell. Biol. 122:53-65). Concurrent with ligand-dependent uptake, we now show that the receptor undergoes ligand-induced ubiquitination, suggesting that receptor ubiquitination may function in the ligand-dependent endocytosis of the a-factor receptor as well as in its constitutive endocytosis. To account for these findings, we propose a model wherein the covalent attachment of ubiquitin to surface receptor triggers endocytic uptake. PMID- 8707847 TI - Xenopus nonmuscle myosin heavy chain isoforms have different subcellular localizations and enzymatic activities. AB - There are two isoforms of the vertebrate nonmuscle myosin heavy chain, MHC-A and MHC-B, that are encoded by two separate genes. We compared the enzymatic activities as well as the subcellular localizations of these isoforms in Xenopus cells. MHC-A and MHC-B were purified from cells by immunoprecipitation with isoform-specific peptide antibodies followed by elution with their cognate peptides. Using an in vitro motility assay, we found that the velocity of movement of actin filaments by MHC-A was 3.3-fold faster than that by MHC-B. Likewise, the Vmax of the actin-activated Mg(2+)-ATPase activity of MHC-A was 2.6 fold greater than that of MHC-B. Immunofluorescence microscopy demonstrated distinct localizations for MHC-A and MHC-B. In interphase cells, MHC-B was present in the cell cortex and diffusely arranged in the cytoplasm. In highly polarized, rapidly migrating interphase cells, the lamellipodium was dramatically enriched for MHC-B suggesting a possible involvement of MHC-B based contractions in leading edge extension and/or retraction. In contrast, MHC-A was absent from the cell periphery and was arranged in a fibrillar staining pattern in the cytoplasm. The two myosin heavy chain isoforms also had distinct localizations throughout mitosis. During prophase, the MHC-B redistributed to the nuclear membrane, and then resumed its interphase localization by metaphase. MHC-A, while diffuse within the cytoplasm at all stages of mitosis, also localized to the mitotic spindle in two different cultured cell lines as well as in Xenopus blastomeres. During telophase both isoforms colocalized to the contractile ring. The different subcellular localizations of MHC-A and MHC-B, together with the data demonstrating that these myosins have markedly different enzymatic activities, strongly suggests that they have different functions. PMID- 8707849 TI - emo-1, a Caenorhabditis elegans Sec61p gamma homologue, is required for oocyte development and ovulation. AB - emo-1(oz1) is a member of a class of hermaphrodite sterile mutations in Caenorhabditis elegans that produce endomitotic oocytes in the gonad arm. Oocytes in emo-1(oz1) mutants exhibit multiple defects during oogenesis. After meiotic maturation, ovulation fails, trapping oocytes in the gonad arm where they become endomitotic. emo-1 encodes a homologue of the Sec61p gamma subunit, a protein necessary for translocation of secretory and transmembrane proteins into the endoplasmic reticulum of yeast and mammalian cells. A putative emo-1 null mutation, oz151, displays embryonic lethality. The oz1 sterile mutation is a transposable element insertion into the emo-1 3' untranslated region that almost completely eliminates germline mRNA accumulation. Genetic mosaic analysis using the oz1 allele indicates that emo-1(+) expression in germ cells is required for fertility. The J67 monoclonal antibody, which recognizes an oocyte surface antigen (Strome, S. 1986. In Gametogenesis and the Early Embryo. J.G. Gall, editor. Alan R. Liss, Inc., New York. 77-95.), does not stain oz1 oocytes, a finding consistent with defective protein transport in the mutant. We propose that the emo-1 gene product acts in the transport of secreted and transmembrane proteins in C. elegans oocytes, and is necessary for both oogenesis and the coupling of ovulation with meiotic maturation. PMID- 8707848 TI - A role for gelsolin in actuating epidermal growth factor receptor-mediated cell motility. AB - Phospholipase C-gamma (PLC gamma) is required for EGF-induced motility (Chen, P., H. Xie, M.C. Sekar, K.B. Gupta, and A. Wells. J. Cell Biol. 1994. 127:847-857); however, the molecular basis of how PLC gamma modulates the actin filament network underlying cell motility remains undetermined. We propose that one connection to the actin cytoskeleton is direct hydrolysis of PIP2 with subsequent mobilization of membrane-associated actin modifying proteins. We used signaling restricted EGFR mutants expressed in receptor-devoid NR6 fibroblast cells to investigate whether EGFR activation of PLC causes gelsolin mobilization from the cell membrane in vivo and whether this translocation facilitates cell movement. Gelsolin anti-sense oligonucleotide (20 microM) treatment of NR6 cells expressing the motogenic full-length (WT) and truncated c'1000 EGFR decreased endogenous gelsolin by 30-60%; this resulted in preferential reduction of EGF (25 nM) induced cell movement by > 50% with little effect on the basal motility. As 14 h of EGF stimulation of cells did not increase total cell gelsolin content, we determined whether EGF induced redistribution of gelsolin from the membrane fraction. EGF treatment decreased the gelsolin mass associated with the membrane fraction in motogenic WT and c'1000 EGFR NR6 cells but not in cells expressing the fully mitogenic, but nonmotogenic c'973 EGFR. Blocking PLC activity with the pharmacologic agent U73122 (1 microM) diminished both this mobilization of gelsolin and EGF-induced motility, suggesting that gelsolin mobilization is downstream of PLC. Concomitantly observed was reorganization of submembranous actin filaments correlating directly with PLC activation and gelsolin mobilization. In vivo expression of a peptide that is reported to compete in vitro with gelsolin in binding to PIP2 dramatically increased basal cell motility in NR6 cells expressing either motogenic (WT and c'1000) or nonmotogenic (c'973) EGFR; EGF did not further augment cell motility and gelsolin mobilization. Cells expressing this peptide demonstrated actin reorganization similar to that observed in EGF-treated control cells; the peptide-induced changes were unaffected by U73122. These data suggest that much of the EGF-induced motility and cytoskeletal alterations can be reproduced by displacement of select actin modifying proteins from a PIP2-bound state. This provides a signaling mechanism for translating cell surface receptor-mediated biochemical reactions to the cell movement machinery. PMID- 8707850 TI - Envoplakin, a novel precursor of the cornified envelope that has homology to desmoplakin. AB - The cornified envelope is a layer of transglutaminase cross-linked protein that is deposited under the plasma membrane of keratinocytes in the outermost layers of the epidermis. We present the sequence of one of the cornified envelope precursors, a protein with an apparent molecular mass of 210 kD. The 210-kD protein is translated from a 6.5-kb mRNA that is transcribed from a single copy gene. The mRNA was upregulated during suspension-induced terminal differentiation of cultured human keratinocytes. Like other envelope precursors, the 210-kD protein became insoluble in SDS and beta-mercaptoethanol on activation of transglutaminases in cultured keratinocytes. The protein was expressed in keratinizing and nonkeratinizing stratified squamous epithelia, but not in simple epithelia or nonepithelial cells. Immunofluorescence staining showed that in epidermal keratinocytes, both in vivo and in culture, the protein was upregulated during terminal differentiation and partially colocalized with desmosomal proteins. Immunogold EM confirmed the colocalization of the 210-kD protein and desmoplakin at desmosomes and on keratin filaments throughout the differentiated layers of the epidermis. Sequence analysis showed that the 210-kD protein is homologous to the keratin-binding proteins desmoplakin, bullous pemphigoid antigen 1, and plectin. These data suggest that the 210-kD protein may link the cornified envelope to desmosomes and keratin filaments. We propose that the 210 kD protein be named "envoplakin." PMID- 8707851 TI - Pantophysin is a ubiquitously expressed synaptophysin homologue and defines constitutive transport vesicles. AB - Certain properties of the highly specialized synaptic transmitter vesicles are shared by constitutively occurring vesicles. We and others have thus identified a cDNA in various nonneuroendocrine cell types of rat and human that is related to synaptophysin, one of the major synaptic vesicle membrane proteins, which we termed pantophysin. Here we characterize the gene structure, mRNA and protein expression, and intracellular distribution of pantophysin. Its mRNA is detected in murine cell types of nonneuroendocrine as well as of neuroendocrine origin. The intron/exon structure of the murine pantophysin gene is identical to that of synaptophysin except for the last intron that is absent in pantophysin. The encoded polypeptide of calculated mol wt 28,926 shares many sequence features with synaptophysin, most notably the four hydrophobic putative transmembrane domains, although the cytoplasmic end domains are completely different. Using antibodies against the unique carboxy terminus pantophysin can be detected by immunofluorescence microscopy in both exocrine and endocrine cells of human pancreas, and in cultured cells, colocalizing with constitutive secretory and endocytotic vesicle markers in nonneuroendocrine cells and with synaptophysin in cDNA-transfected epithelial cells. By immunoelectron microscopy, the majority of pantophysin reactivity is detected at vesicles with a diameter of < 100 nm that have a smooth surface and an electron-translucent interior. Using cell fractionation in combination with immunoisolation, these vesicles are enriched in a light fraction and shown to contain the cellular vSNARE cellubrevin and the ubiquitous SCAMPs in epithelial cells and synaptophysin in neuroendocrine or cDNA transfected nonneuroendocrine cells and neuroendocrine tissues. Pantophysin is therefore a broadly distributed marker of small cytoplasmic transport vesicles independent of their content. PMID- 8707852 TI - Regulation of tyrosinase gene expression by cAMP in B16 melanoma cells involves two CATGTG motifs surrounding the TATA box: implication of the microphthalmia gene product. AB - In melanocytes and in melanoma cells, upregulation of melanogenesis, by cAMP elevating agents, results from a stimulation of tyrosinase activity that has been ascribed to an increase in tyrosinase protein and messenger amount. However, the mechanism by which cAMP elevating agents increase tyrosinase mRNA remains to be elucidated. In this study, using a luciferase reporter plasmid containing the 2.2 kb fragment 5' of the transcriptional start site of the mouse tyrosinase gene, we showed that cAMP elevating agents lead to a strong stimulation (20-fold) of transcriptional activity of the tyrosinase promoter. Deletions and mutations in the mouse tyrosinase promoter showed that the M-box 70-bp upstream from the TATA box and the E-box located downstream the TATA-box, near to the initiator site, are involved in the regulation of the tyrosinase promoter activity by cAMP. Additionally, we showed that microphthalmia, a b-HLH transcription factor associated with pigmentation disorders in mouse, binds to these regulatory elements and modulates the transcriptional activity of the tyrosinase promoter. Since cAMP stimulates the binding of microphthalmia to the M-box and to the E box; it is tempting to propose that microphthalmia, through its interaction with cis-acting elements surrounding the TATA-box, plays a key role in the regulation of the mouse tyrosinase gene expression by cAMP. PMID- 8707853 TI - Induction of neuronal apoptosis by camptothecin, an inhibitor of DNA topoisomerase-I: evidence for cell cycle-independent toxicity. AB - Camptothecin is an S-phase-specific anticancer agent that inhibits the activity of the enzyme DNA topoisomerase-I (topo-I). Irreversible DNA double-strand breaks are produced during DNA synthesis in the presence of camptothecin, suggesting that this agent should not be toxic to nondividing cells, such as neurons. Unexpectedly, camptothecin induced significant, dose-dependent cell death of postmitotic rat cortical neurons in vitro; astrocytes were more resistant. Aphidicolin, an inhibitor of DNA polymerase alpha, did not prevent camptothecin induced neuronal death, while death was prevented by actinomycin D and 5,6 dichloro-1-beta-D-ribofuranosyl benzimidazole as well as cycloheximide and anisomycin, inhibitors of RNA and protein synthesis, respectively. Camptothecin induced neuronal death was apoptotic, as characterized by chromatin condensation, cytoplasmic shrinking, plasma membrane blebbing, and fragmentation of neurites. DNA fragmentation was also confirmed by the use of the in situ DNA end labeling assay. In addition, aurintricarboxylic acid, an inhibitor of the apoptotic endonuclease, partially protected against camptothecin-induced neuronal death. The toxicity of stereoisomers of a camptothecin analogue was stereospecific, demonstrating that toxicity was a result of inhibition of topo-I. The difference in sensitivity to camptothecin between neurons and astrocytes correlated with their transcriptional activity and level of topo-I protein expression. These data indicate important roles for topo-I in postmitotic neurons and suggest that topo I inhibitors can induce apoptosis independent of DNA synthesis. We suggest a model based on transcriptionally mediated DNA damage, a novel mechanism of action of topo-I poisons. PMID- 8707854 TI - Identification of stromal cell products that interact with pre-B cells. AB - Our understanding of lympho-hematopoietic microenvironments is incomplete, and a new cloning strategy was developed to identify molecules that bind to B lineage lymphocyte precursors. A cell sorting procedure was used for initial enrichment of cDNAs from stromal cell mRNA that contained signal sequences and were therefore likely to encode transmembrane or secreted proteins. A second step involved expression of the library as soluble Ig fusion proteins. Finally, pools representing these proteins were screened for the ability to recognize pre-B cells. This approach resulted in the cloning of biglycan, syndecan 4, collagen type I, clusterin, matrix glycoprotein sc1, osteonectin, and one unknown molecule (designated SIM). The full-length cDNA of SIM revealed that it is a type I transmembrane protein, and its intracellular domain has weak homology with myosin heavy chain and related proteins. Staining of established cell lines and freshly isolated hematopoietic cells with the Ig fusion proteins revealed distinct patterns of reactivity and differential dependence on divalent cations. Biglycan , sc1-, and SIM-Ig fusion proteins selectively increased interleukin 7-dependent proliferation of pre-B cells. Overexpression of the entire SIM protein affected the morphology of 293T cells, while expression of just the extracellular portion was without effect. Thus, a series of stromal cell surface molecules has been identified that interact with blood cell precursors. Three of them promoted the survival and/or proliferation of pre-B cells in culture, and all merit further study in relation to lympho-hematopoiesis. PMID- 8707855 TI - FGF-1-dependent proliferative and migratory responses are impaired in senescent human umbilical vein endothelial cells and correlate with the inability to signal tyrosine phosphorylation of fibroblast growth factor receptor-1 substrates. AB - Senescent cells do not proliferate in response to exogenous growth factors, yet the number and affinity of growth factor receptors on the cell surface appear to be similar to presenescent cell populations. To determine whether a defect in receptor signaling exists, we analyzed human umbilical vein endothelial cells (HUVEC) since HUVEC growth is absolutely dependent upon the presence of FGF. We report that in both presenescent and senescent HUVEC populations, FGF-1 induces the expression of cell cycle-specific genes, suggesting that functional FGF receptor (FGFR) may exist on the surface of these cells. However, the tyrosine phosphorylation of FGFR-1 substrates, Src and cortactin, is impaired in senescent HUVEC, and only the presenescent cell populations exhibit a FGF-1-dependent Src tyrosine kinase activity. Moreover, we demonstrate that senescent HUVEC are unable to migrate in response to FGF-1, and these data correlate with an altered organization of focal adhesion sites. These data suggest that the induction of gene expression is insufficient to promote a proliferative or migratory phenotype in senescent HUVEC and that the attenuation of the FGFR-1 signal transduction pathway may be involved in the inability of senescent HUVEC to proliferate and/or migrate. PMID- 8707858 TI - Regulation of changes in cytosolic Ca2+ and Na+ concentrations in rat submandibular gland acini exposed to carbachol and ATP. AB - The relationship between cytosolic concentrations of Ca2+ (Ca2i) and Na+ (Na+i) were studied in preparations of rat submandibular and pancreatic acini loaded with the Ca(2+)-sensitive dye Fura-2 or the Na(+)-sensitive dye SBFI. Pancreatic acini showed no changes in Na+i during either transient or persistent changes in Ca2+i. Increases in Ca2+i produced by exposure of submandibular gland acini to carbachol, a muscarinic cholinergic agonist, were followed by an increase in Na+i after a delay of 5-10 s. When Ca2+ stores were mobilized without Ca2+ influx Na+i also increased, but in acini loaded with BAPTA, a nonfluorescent Ca2+ chelator, the transient increase in Ca2+ caused by mobilization of stored Ca2+ was virtually abolished, as was the increase in Na+i. In the presence of inomycin, increases in Ca2+i were followed by increases in Na+i. Ca(2+)-dependent increases in Na+i were abolished in Na(+)-free buffer and by the presence of furosemide, a blocker of Na(+)-K(+)-2Cl- cotransport. In other studies, extracellular ATP (ATPo) produced an increase in Ca2+i and Na+i. The steady-state increase in Ca(i)2+ was reduced by increasing extracellular Na+ concentrations (Na+o in dose dependent fashion (IC50 = 16.4 +/- 4.7 mM Na+). Likewise, increasing Na+o reduced ATPo-stimulated 45Ca2+ uptake at steady state (IC50 = 15.8 +/- 9.2 mM Na+). Changing Na+o had no effect on carbachol-stimulated increases in Ca2+i. We conclude that, in rat submandibular gland acini, ATPo promotes an increase in Ca2+i and Na+i via a common influx pathway and that, under physiologic conditions, Na+ significantly limits the ATPo-stimulated increase in Ca2+i. In the presence of carbachol, however, Na+i rises in Ca2+i-dependent fashion in submandibular gland acini via stimulation of Na(+)-K(+)-2Cl- cotransport. PMID- 8707857 TI - Regulated binding of PTP1B-like phosphatase to N-cadherin: control of cadherin mediated adhesion by dephosphorylation of beta-catenin. AB - Cadherins are a family of cell-cell adhesion molecules which play a central role in controlling morphogenetic movements during development. Cadherin function is regulated by its association with the actin containing cytoskeleton, an association mediated by a complex of cytoplasmic proteins, the catenins: alpha, beta, and gamma. Phosphorylated tyrosine residues on beta-catenin are correlated with loss of cadherin function. Consistent with this, we find that only nontyrosine phosphorylated beta-catenin is associated with N-cadherin in E10 chick retina tissue. Moreover, we demonstrate that a PTP1B-like tyrosine phosphatase associates with N-cadherin and may function as a regulatory switch controlling cadherin function by dephosphorylating beta-catenin, thereby maintaining cells in an adhesion-competent state. The PTP1B-like phosphatase is itself tyrosine phosphorylated. Moreover, both direct binding experiments performed with phosphorylated and dephosphorylated molecules, and treatment of cells with tyrosine kinase inhibitors indicate that the interaction of the PTP1B like phosphatase with N-cadherin depends on its tyrosine phosphorylation. Concomitant with the tyrosine kinase inhibitor-induced loss of the PTP1B-like phosphatase from its association with N-cadherin, phosphorylated tyrosine residues are retained on beta-catenin, the association of N-cadherin with the actin containing cytoskeleton is lost and N-cadherin-mediated cell adhesion is prevented. Tyrosine phosphatase inhibitors also result in the accumulation of phosphorylated tyrosine residues on beta-catenin, loss of the association of N cadherin with the actin-containing cytoskeleton, and prevent N-cadherin mediated adhesion, presumably by directly blocking the function of the PTP1B-like phosphatase. We previously showed that the binding of two ligands to the cell surface N-acetylgalactosaminylphosphotransferase (GalNAcPTase), the monoclonal antibody 1B11 and a proteoglycan with a 250-kD core protein, results in the accumulation of phosphorylated tyrosine residues on beta-catenin, uncoupling of N cadherin from its association with the actin containing cytoskeleton, and loss of N-cadherin function. We now report that binding of these ligands to the GalNAcPTase results in the absence of the PTP1B-like phosphatase from its association with N-cadherin as well as the loss of the tyrosine kinase and tyrosine phosphatase activities that otherwise co-precipitate with N-cadherin. Control antibodies and proteoglycans have no such effect. This effect is similar to that observed with tyrosine kinase inhibitors, suggesting that the GalNAcPTase/proteoglycan interaction inhibits a tyrosine kinase, thereby preventing the phosphorylation of the PTP1B-like phosphatase, and its association with N-cadherin. Taken together these data indicate that a PTP1B-like tyrosine phosphatase can regulate N-cadherin function through its ability to dephosphorylate beta-catenin and that the association of the phosphatase with N cadherin is regulated via the interaction of the GalNAcPTase with its proteoglycan ligand. In this manner the GalNAcPTase-proteoglycan interaction may play a major role in morphogenetic cell and tissue interactions during development. PMID- 8707856 TI - Control of adhesion-dependent cell survival by focal adhesion kinase. AB - The interactions of integrins with extracellular matrix proteins can activate focal adhesion kinase (FAK) and suppress apoptosis in normal epithelial and endothelial cells; this subset of apoptosis has been termed "anoikis." Here, we demonstrate that FAK plays a role in the suppression of anoikis. Constitutively activated forms of FAK rescued two established epithelial cell lines from anoikis. Both the major autophosphorylation site (Y397) and a site critical to the kinase activity (K454) of FAK were required for this effect. Activated FAK also transformed MDCK cells, by the criteria of anchorage-independent growth and tumor formation in nude mice. We provide evidence that this transformation resulted primarily from the cells' resistance to anoikis rather than from the activation of growth factor response pathways. These results indicate that FAK can regulate anoikis and that the conferral of anoikis resistance may suffice to transform certain epithelial cells. PMID- 8707859 TI - Changes in mitochondrial activity during avian myoblast differentiation: influence of triiodothyronine or v-erb A expression. AB - Numerous data suggest that mitochondrial activity is involved in the regulation of cell growth and differentiation. Therefore, we have studied the changes in mitochondrial activity in avian myoblast cultures (QM7 line) undergoing differentiation or in BrdU-treated, differentiation-deficient cells. As we have previously shown that triiodothyronine and v-erb A expression stimulate myogenic differentiation, we have also observed their influence upon mitochondrial activity. Comparison of control and BrdU-treated myoblasts indicated that precocious differentiation events were associated with a stimulation of citrate synthase and cytochrome oxidase activities. They also induced a transient decrease in mitochondrial membrane potential assessed by rhodamine 123 uptake. In control myoblasts, a general stimulation of mitochondrial activity was recorded at cell confluence, prior to terminal differentiation. These events did not occur in BrdU-treated myoblasts, thus indicating that they were tightly linked to myoblast commitment. Whereas no significant triiodothyronine influence could be detected upon mitochondrial activity, we observed that v-erb A expression significantly depresses the mitochondrial membrane potential in control myoblasts. This action was not observed in BrdU-treated myoblasts, thus suggesting that it involves an indirect pathway linked to differentiation. Moreover, the oncoprotein abrogated the decrease in E2-PDH subunit level observed at cell confluence. These data underline that changes in mitochondrial activity occurred prior to myoblast terminal differentiation and could be involved in the processes regulating myogenesis. In addition, they provide the first evidence that the v-erb A oncoprotein influences mitochondrial activity. PMID- 8707860 TI - Intracellular location of cysteine transport activity correlates with productive processing of antigen disulfide. AB - Activation of CD4+ T cells requires processing of exogenous protein antigens by antigen-presenting cells (APC). A macrophage hybridoma and B cell lymphoma were comparable in their ability to process hen egg lysozyme (HEL), which involves reduction of its disulfide bonds. The intracellular levels of cysteine and glutathione, major physiological thiols, based on protein content were similar within these cell lines. In addition, the cysteine transport pathway in viable cells was assessed by 35S-cystine uptake. For macrophages, the majority of the radioactivity resided in high density subcellular fractions of Percoll gradients that comigrated with lysosomal beta-galactosidase (beta-gal). Besides the lysosomes, low density fractions cosedimenting with endosomes incorporated the radiolabel in the B cells. Both peaks of radioactivity disappeared when the B cells were incubated with unlabeled carboxymethyl-cysteine (CM-cysteine), a specific competitor of the plasma membrane CG transport system. The distinct gradient profiles of radiolabel uptake in the cells correlated with a difference in their capacity to process the transferrin-lysozyme conjugate (TF-HEL). TF-HEL was significantly more stimulatory than HEL in inducing a HEL-specific T cell response with the B cells as the APC. However, the potencies of TF-HEL and HEL were similar when the macrophages were the APC. Thus, the intracellular location of cysteine transport activity may be cell lineage-dependent, and its presence may, in part, determine whether an organelle is a productive site of processing antigens with disulfide bonds that is necessary for CD4+ cell activation. PMID- 8707861 TI - C-myc expression affects proliferation but not terminal differentiation or survival of explanted erythroid progenitor cells. AB - The expression of c-myc was analyzed in murine and human erythroblasts throughout their differentiation in vitro into reticulocytes. The murine cells were splenic erythroblasts from animals infected with the anemia strain of Friend virus (FVA cells). In FVA cells cultured without EPO, the c-myc mRNA and protein levels decrease sharply within 3 to 4 h, showing that continual EPO stimulation is required to maintain c-myc expression. When cultured with EPO, the c-myc mRNA level of FVA cells is raised within 30 min of exposure. The c-myc mRNA and protein reach maxima at 1 to 3 h, then decline slowly to very low levels by 18 h. In contrast, c-fos and c-jun mRNA levels are not regulated by EPO in FVA cells. The human cells analyzed were colony-forming units-erythroid, CFU-E, derived in vitro by the culture of peripheral blood burst-forming units-erythroid (BFU-E). When grown in EPO and insulin-like growth factor 1 (IGF-1) these cells differentiate into reticulocytes over 6 days rather than the 2 days required for murine cells, but the c-myc mRNA kinetics and response to EPO parallel those of mouse cells at similar stages of differentiation. Both IGF-1 and c-kit ligand (SCF) cause an additive increase in c-myc mRNA in human CFU-E in conjunction with EPO. These additive effects suggest that EPO, IGF-1, and SCF affect c-myc mRNA accumulation by distinct mechanisms. Addition of an antisense oligonucleotide to c-myc in cultures of human CFU-E specifically inhibited cell proliferation but did not affect erythroid cell differentiation or apoptosis. When human cells were grown in high SCF concentrations, an environment which enhances proliferation and retards differentiation, antisense oligonucleotide to c-myc strongly inhibited proliferation, but such inhibition did not induce differentiation. This latter result indicates that differentiation requires signals other than depression of c Myc and resultant depression of proliferation. PMID- 8707862 TI - Regulation of protein abundance in pluripotent cells undergoing commitment to the neural lineage. AB - The P19 cell line is a widely studied model of neural differentiation When pluripotent P19 cells are cultured as aggregates in the presence of retinoic acid for 4 days, the cells commit to the neural fate, but have not yet undergone overt differentiation. Two-dimensional polyacrylamide gel electrophoresis was used to analyze cellular protein expression during this induction. Approximately 500 abundant polypeptides were analyzed. Seventeen polypeptides were upregulated during induction; several of these were significantly regulated 48 h after the addition of retinoic acid. No downregulations were observed. Fifteen of the 17 polypeptides continued to be expressed throughout terminal differentiation. The upregulation of 14 of the 17 polypeptides requires both retinoic acid and aggregation, which alone do not induce neural differentiation. Furthermore, these regulated polypeptides are expressed in neural tissue, suggesting they are associated with neural function in vivo. Embryonic stem cells, a totipotent line, also neurally differentiate in response to retinoic acid and aggregation. Comparison of embryonic stem cells to P19 cells shows that the two systems regulate a similar set of polypeptides and are thus likely to utilize a similar pathway. These studies are a step toward determining the full extent of regulation involved in the commitment of pluripotent cells to the neural fate. PMID- 8707864 TI - Cleavage of the transferrin receptor is influenced by the composition of the O linked carbohydrate at position 104. AB - A soluble form of the human transferrin receptor (TfR) resulting from proteolytic cleavage at Arg 100 has been measured in human blood. In tissue culture cells elimination of the O-linked carbohydrate at Thr 104, four amino acids from the cleavage site, results in enhanced cleavage of the TfR (Rutledge et al., 1994, Blood, 83:580-586). In the present set of studies, the influence of amino acid substitution and the composition of the oligosaccharide at amino acid 104 on the cleavage of the TfR was examined. Site-directed mutagenesis was used to generate six different amino acids at position 104 which varied in size and charge. Measurement of the soluble TfR in the conditioned medium of the transfected cells of each mutant TfR showed that the large and charged side chains inhibited TfR cleavage the most. Otherwise the properties of the mutant TfRs were indistinguishable from the wild-type TfR in that the affinity of transferrin for these receptors, the extent of disulfide bond formation of the TfRs, and the proportion of TfRs at the cell surface were similar to that of the wild-type TfR. Removal of the sialic acid component of the carbohydrate from wild-type TfR by treatment of live cells with neuraminidase enhances TfR cleavage. Expression of wild-type TfR in CHO IdlD cells (a glycosylation defective cell line) also shows enhanced cleavage under conditions that produce truncated or no O-linked carbohydrates. Treatment of IdlD cells with neuraminidase reveals that the sialic acid of the O-linked carbohydrate protects against TfR cleavage, whereas the core sugars Gal-NAc and Gal do not protect as much. These results show that the terminal charged sialic acid residues are important for protection from proteolytic cleavage and suggest that cleavage could be regulated in the cell by removal of all or part of the carbohydrate. PMID- 8707863 TI - Calcium regulated chloride permeabilities in primary cultures of rabbit colonocytes. AB - To determine if calcium-dependent secretagogues directly act on epithelial cells to elicit Cl- secretion, their effects on Cl- transport and intracellular Ca(2+) concentrations ([Ca2+]i) were determined in primary cultures of rabbit distal colonic crypt cells. The Cl- sensitive fluorescent probe, 6-methoxyquinolyl acetoethyl ester, MQAE and the Ca(2+)-sensitive fluorescent probe, fura-2AM were used to assess Cl- transport and [Ca2+]i, respectively. Basal Cl- transport (0.274 +/- 0.09 mM/sec) was inhibited significantly by the Cl- channel blocker diphenylamine-2-carboxylate (DPC, 50 microM, 0.068 +/- 0.02 mM/sec; P < 0.001) and the Na+/K+/ 2Cl- cotransport inhibitor furosemide (1 microM, 0.137 +/- 0.04 mM/sec; P < 0.01). Ion substitution studies using different halides revealed the basal influx to be l- > F- > or = Cl- > Br-. DPC inhibited l- influx by approximately 50%, F- influx by 80%, Cl-influx by 85%, and Br- influx by 90%. Furosemide significantly inhibited influx of Br- (84%) and Cl- (81%) but not of F and l-. The effects of agents known to alter biological response by increasing [Ca2+]i in other epithelial systems were used to stimulate Cl- transport. Cl- influx in mM/second was stimulated by 1 microM histamine (0.58 +/- 0.05), 10 microM neurotensin (2.07 +/- 0.32), 1 microM serotonin (1.63 +/- 0.28), and 0.1 microM of the Ca2+ ionophore A23187 (2.05 +/- 0.40). The Cl- permeability stimulated by neurotensin, serotonin, and A23187 was partially blocked by DPC or furosemide added alone or in combination. Histamine-induced Cl- influx was significantly inhibited by only furosemide. Indomethacin blocked histamine stimulated Cl- permeability but had no effect on the actions of the other agents. These studies, focusing on isolated colonocytes without the contribution of submucosal elements, reveal that (1) histamine stimulates Cl- transport by activating the Na+/K+/2Cl- cotransporter via a cyclooxygenase-dependent pathway; (2) neurotensin, serotonin, and A23187 activate both Cl- channels and the cotransporter, and their actions are cyclooxygenase-independent. PMID- 8707865 TI - Effects of basic fibroblast growth factor on human microvessel endothelial cell migration on collagen I correlates inversely with adhesion and is cell density dependent. AB - Angiogenesis, new vessel growth from existing vessels, is critical to tissue development and healing. Much is known about the molecular and cellular elements of angiogenesis, such as the effects of growth factors and matrix molecules on proliferation and migration. However, it is not clear how these elements are coordinated to produce specific microvascular beds. To address this, the effects of basic fibroblast growth factor (bFGF) on beta 1 integrin-mediated adhesion relative to migration in human microvessel endothelial cells (HMVEC) was examined. Using two assays of migration that differ in the density of cells being examined, bFGF stimulated single cell migration and reduced cell migration from a confluent monolayer on collagen I. Adhesion to collagen I of HMVEC treated at low density (2-4 x 10(4) cells/cm2) with bFGF for 22 h was reduced, while bFGF increased cell adhesion of HMVEC treated at high density (6-8 x 10(4) cells/cm2). Adhesion of both bFGF-treated and untreated HMVEC was mediated by the beta 1 integrin matrix receptor. Basic FGF treatment did not significantly alter surface expression of the beta 1 integrin subunit. Reduction in bFGF-mediated adhesion correlated with delayed cell spreading and altered organization of beta 1 integrin into substrate contacts. Thus, integrin-mediated cell adhesion in microvessel endothelial cells is sensitive to regulation by a growth factor. Furthermore, the nature of the response to this signal depends on another cell regulator, cell density. In addition, modulation of cell adhesion by a growth factor may be a central regulatory feature in controlling endothelial cell migration. PMID- 8707866 TI - Membrane phase transition of intact human platelets: correlation with cold induced activation. AB - Using Fourier transform infrared spectroscopy (FTIR), we have determined the phase transition temperature (Tm) of lipids in intact human platelets and have shown that it occurs between 15 and 18 degrees C, the temperature at which cold activation of platelets has previously been reported (Zucker and Borrelli, 1954, Blood, 28:602-608; White and Krivit, 1967, Blood, 30:625-635). The temperature at which the platelets pass through Tm is highly correlated with initial platelet shape change. However, shape change continues after the cells have passed through the phase transition. Cold-induced activation has previously prevented long-term storage of platelets at 4 degrees C. Antifreeze glycoproteins (AFGPs) isolated from polar fishes previously have been used to prevent ice crystal growth during freezing of tissues as well as leakage of solutes from liposomes as they were chilled through their Tm. We sought to determine if these AFGPs were able to stabilize platelets for long-term storage at 4 degrees C. Incubating platelets with antifreeze glycoproteins during long-term storage and rapid rewarming to 37 degrees C abrogated granule secretion associated with cold activation in a dose dependent manner. This work suggests that AFGPs may be a possible solute for use in long-term low temperature storage of platelets. PMID- 8707867 TI - Adenosine-mediated inhibition of casein production by mouse mammary glands in culture. AB - The present study was carried out to examine whether activation of adenosine receptors by adenosine analogues will affect casein production by mouse mammary epithelial cells. The morphogenesis and functions of epithelial tissue in the mammary gland are influenced by their surrounding adipocytes. Adipocytes are known to release adenosine into the extracellular fluid which can modulate cyclic AMP levels in surrounding cells through binding to their adenosine receptors. To examine a possible paracrine effect of adenosine, the modulation of casein production in mammary explant culture and mammary epithelial cell (MEC) culture by adenosine receptor agonists has been investigated. We have observed that activation of the A1-adenosine receptor subtype in mammary tissue by an adenosine analogue (-)N6-(R-phenyl-isopropyl)-adenosine (PIA) raised cAMP levels. PIA and another adenosine receptor agonist, isobutylmethylxanthine (IBMX), inhibited casein accumulation both in explants and in MEC cultures in the presence of lactogenic hormones, which suggests that PIA or adenosine can act directly on the epithelial cells. This inhibition does not appear to be caused by elevation of cAMP levels or phosphodiesterase activity. The inhibition of intracellular casein accumulation by PIA and IBMX in explant cultures can be reversed via treatment of pertussis toxin which is known to ADP-ribosylate GTP-binding G alpha i-proteins, indicating that a Gi-protein-dependent pathway may be involved in this inhibition. The results also suggest that local accumulation of adenosine in the extracellular fluids of mammary glands is likely to inhibit the lactogenic response of mammary epithelial cells. PMID- 8707868 TI - Extracellular matrix is a source of mitogenically active platelet-derived growth factor. AB - Platelet-derived growth factor (PDGF) is a chemotactic and mitogenic agent for fibroblasts and smooth muscle cells and plays a key role in the development of atherosclerotic lesions. PDGF is produced by a number of normal and transformed cell types and occurs as homo- or heterodimers of A and B polypeptide chains. Using Chinese hamster ovary (CHO) cells transfected with various forms of PDGF, we have previously shown that PDGF A(s) (short splice version) is secreted, PDGF A(l) (long splice version) predominantly extracellular matrix-associated, and PDGF B divided between medium, cells, and matrix. In the present study we have demonstrated the mitogenic activity of matrix-localized PDGF in artificial and more physiologically relevant models by culturing Balb/c-3T3 cells (3T3), human foreskin fibroblasts (HFF), and rabbit aortic smooth muscle cells (SMC) on extracellular matrix (ECM) laid down by PDGF-expressing CHO cells and human umbilical vein endothelial cells (HUVEC). These cells responded to the local growth stimulus of PDGF-containing CHO ECM and HUVEC ECM. We showed that 3T3 cells required proteolytic activity to utilize matrix-localized PDGF, as aprotinin and epsilon-ACA inhibited growth and 3T3 cells were shown to possess plasminogen activator activity. HFF and SMC did not appear to require proteolytic activity (including metalloproteinase and serine protease activity) as a prerequisite for mitogenesis but were able to access immobilized PDGF by contact with the matrix. An understanding of the mechanisms whereby the utilization of stored PDGF is controlled in situations of excessive cellular proliferation will aid in the development of therapy for these conditions. PMID- 8707869 TI - Cleavage of the transferrin receptor by human granulocytes: differential proteolysis of the exosome-bound TFR. AB - In contrast with other mammalian granulocytes, human granulocytes rapidly cleave the transferrin receptor (TFR) from sheep exosomes. Proteolysis of TFR from exosomes is more rapid and more extensive than that from the sheep reticulocyte cell surface itself, although little difference in cleavage is seen when immunoprecipitates or when immobilized, solubilized receptors from the two sources are compared. Circulating exosomes but not the plasma membrane fraction from seven species of immature red cells or erythropoietic cells show the presence of a peptide of approximately 18 kD recognized by an antibody to the cytoplasmic domain of the TFR. This 18 kD peptide is virtually absent from the corresponding cellular plasma membranes including human reticulocyte membranes. Taken together, the data are consistent with the conclusion that the exosomes released to the circulation from maturing red cells are the principal source of the soluble, circulating, truncated TFR. The granulocyte protease appears to be present on the cell surface and not released into the medium after short (30 min) periods of incubation at 37 degrees C. The protease is inhibited by PMSF but only at high (1 mM) concentrations. Using sheep exosome bound-TFR as substrate, human granulocytes exceed other granulocytes in their capacity to cleave TFR, suggesting that this may be a key factor for the prominent amount of circulating, soluble receptor found in human sera during periods of elevated reticulocyte levels. Human exosomes, unlike those from other species, contain little native size TFR. Truncated receptor containing the cytoplasmic domain being predominant in human exosomes. PMID- 8707870 TI - Hypoxia impairs cell fusion and differentiation process in human cytotrophoblast, in vitro. AB - During human pregnancy, the trophoblast develops from differentiation of cytotrophoblast cells into an endocrine active syncytiotrophoblast. In culture, isolated mononuclear cytotrophoblasts aggregate and then fuse to form a syncytium, reproducing the in vivo process. In this study, we examined the effect of low oxygen tension (approximately 9%, hypoxia) compared to standard conditions (approximately 19% oxygen, normoxia) on these cellular events. Under hypoxia, syncytial formation was less frequently observed, cell staining and electron microscopy revealed that cytotrophoblasts remain aggregated, with a positive proliferative cell nuclear antigen (PCNA) immunostaining. Desmoplakin and E cadherin, both known to disappear with cytotrophoblast fusion, showed persistent expression in hypoxic cells after 3 days of culture. In contrast, the expression of actin and ezrin, two cytoskeletal proteins, was unchanged. hCG secretion and hPL expression were both decreased in hypoxic cells, reflecting a reduced syncytial formation. Thus, on day 3, the mean values for hCG secretion were 1,100 +/- 155 and 289 +/- 26 mlU/mL in normoxic and hypoxic conditions, respectively. The reduced cell fusion process as well as hCG secretion and hPL expression under hypoxia were reversed by reoxygenation of the cells. We conclude that under hypoxia, the formation of functional syncytiotrophoblast is impaired due to a defect in the cytotrophoblast fusion process. This may explain the observation of a higher number of cytotrophoblast cells and a reduced syncytial layer in placentas of some pathological pregnancies. PMID- 8707871 TI - Hypoxia signalling in the control of erythropoietin gene expression in rat hepatocytes. AB - This study aimed to investigate the sequence of events involved in the stimulation of erythropoietin (EPO) gene expression by hypoxia in hepatocytes. To this end, primary cultures of rat hepatocytes were kept at either high (40% O2) or low (3% O2) oxygen tensions for 2.5 h. Hypoxia increased EPO mRNA about fifteen-fold, whilst the divalent cation cobalt (50-100 microM) or the iron chelator desferrioxamine (10-200 microM) did not increase EPO mRNA levels. Addition of hydrogen peroxide (100-500 microM) to the culture medium did also not change EPO mRNA levels at high or low oxygen tension. Addition of catalase (50 200 micrograms/ml) to the culture medium resulted in a lower level of hypoxia induced EPO mRNA. Inhibition of protein synthesis by cycloheximide (100 microM) completely abolished the increase of EPO mRNA in response to hypoxia. Hypoxia but not cobalt increased the appearance of the hypoxia-inducible factor 1 (HIF-1), and this increase was blunted by cycloheximide. Taken together, these findings suggest that a classic heme protein and a related oxygen-dependent production of oxygen radicals is less likely to be involved in the regulation of the EPO gene by oxygen in hepatocytes. On the other hand, intact protein synthesis is an absolute requirement for the hypoxia-induced appearance of HIF-1 and for hypoxia stimulated expression of the EPO gene in hepatocytes. PMID- 8707872 TI - Murine interleukin-11 (IL-11) is expressed at high levels in the hippocampus and expression is developmentally regulated in the testis. AB - IL-11, derived from a bone marrow stromal cell line, has pleiotropic effects on both hematopoietic cells and nonhematopoietic cells. However, no previous studies have systematically addressed expression of IL-11 in primary tissues in vivo and the relationship of IL-11 tissue specific gene expression and function of IL-11 is not clear. In the present study, we examined constitutive IL-11 expression in various murine adult tissues in vivo. IL-11 mRNA is expressed in a wide range of normal tissues (including hematopoietic organs) at levels only detected by RT PCR. IL-11 protein was detected in brain and testis by Western blot analysis. The in vivo cellular distribution of IL-11 expression was examined by in situ hybridization. In brain, IL-11 message is distributed in granular layer dentate gyrus and pyramidal cell layers of hippocampus. IL-11 is also expressed in anterior horn cells and lateral column neuronal cells of the spinal cord. In testis, IL-11 mRNA is expressed in round spermatids at stage VI-IX seminiferous tubules. IL-11 expression in testis is restricted to developing spermatogonia and is developmentally regulated, since no expression is seen in mice genetically deficient in germ cells and in mice prior to sexual maturation. These expression data correlate with functional data demonstrating that IL-11 stimulates proliferation in vitro of a hippocampus neuronal progenitor cell line and administration of IL-11 in vivo accelerates recovery of spermatogenesis after cytotoxic therapy. These studies suggest that IL-11 may be an important regulator in neural and testicular function. PMID- 8707873 TI - Turnover of the cystic fibrosis transmembrane conductance regulator (CFTR): slow degradation of wild-type and delta F508 CFTR in surface membrane preparations of immortalized airway epithelial cells. AB - The protein product of the cystic fibrosis (CF) gene, termed the cystic fibrosis transmembrane conductance regulator (CFTR), is known to function as an apical chloride channel at the surface of airway epithelial cells. It has been proposed that CFTR has additional intracellular functions and that there is altered processing of mutant forms. In examining these functions we found a stable form of CFTR with slow turnover in surface membrane preparations from CF and non-CF immortalized airway epithelial cell lines. The methods used to study the turnover of CFTR were pulse/chase experiments utilizing saturation labeling of [35S] Met with chase periods of 5-24 h in the presence of 8 mM Met and cell fractionation techniques. Preparations of morphologically identifiable surface membranes were compared to total cell membrane preparations containing intracellular membranes. Surface membrane CFTR had lower turnover defined by pulse/chase ratios than that of the total cell membrane preparations. Moreover, mutant CFTR was stable in the surface membrane fraction with little degradation even after a 24 h chase, whereas wild-type CFTR had a higher pulse/chase ratio at 24 h. In the presence of 50 microM castanospermine, which is an inhibitor of processing alpha glucosidases, a more rapid turnover of mutant CFTR was found in the total cell membrane preparation, whereas wild-type CFTR had a lower response. The results are compatible with a pool of CFTR in or near the surface membranes which has an altered turnover in CF and a glycosylation-dependent alteration in the processing of mutant CFTR. PMID- 8707874 TI - In vivo effects of vitamin D on the proliferation and differentiation of rat keratinocytes. AB - The hormonal form of vitamin D appears to be a physiological regulator of the epidermogenesis. While its differentiation-promoting effect is well accepted, there are conflicting reports of its action on keratinocyte proliferation. This study evaluates the specific changes induced by vitamin D treatment in the epidermis of rats nutritionally deprived of vitamin D by cell size analysis, acridine orange flowcytometry, and the immunohistochemical detection of proteins related to the different stages of differentiation (epidermal calcium binding protein and suprabasal keratins recognized by KL1 antibody) The total keratinocyte and isolated keratinocyte subpopulations were studied. Vitamin D deficiency was associated in the total population with a lower percentage of actively proliferating cells and with a lack of differentiation markers. Study of the isolated cell populations demonstrated, however, that small cells were actively proliferating, whereas they were mainly in the resting stage in the normal epidermis. Treatment with vitamin D dramatically increased cell proliferation and stimulated the appearance of differentiation markers. Some of the observed effects, such as an increase in proliferation and the appearance of epidermal calcium binding protein, were due to the normalisation of the vitamin D deficiency-induced hypocalcemia, whereas the expression of suprabasal keratins was directly dependent on vitamin D. We conclude that the action of vitamin D on the epidermis is associated with increases in both proliferation and differentiation of keratinocytes. Vitamin D itself and its resulting action on calcium homeostasis appear to contribute to the observed effects. PMID- 8707875 TI - Basic fibroblast growth factor inhibits osteoclast-like cell formation. AB - Basic fibroblast growth factor (bFGF) inhibited osteoclast-like cell formation in co-cultures of mouse bone marrow cells either with the mouse stromal cell line, ST2, or with primary osteoblastic cells. Basic FGF significantly inhibited the osteoclast-like cell formation, induced by 1 alpha,25-dihydroxyvitamin D3[1 alpha, 25(OH)2D3] when the cytokine was added to the culture, at an intermediate stage, suggesting that bFGF inhibits the differentiation of the osteoclast progenitors. With regard to target cells, bFGF directly affected ST2; it increased [3H] thymidine uptake and decreased the number of alkaline phosphatase positive cells. In contrast, bFGF had no inhibitory effect on the colony formation of bone marrow cells induced by macrophage colony stimulating factor in methylcellulose culture. In addition, ST2 cells treated with bFGF produced similar amounts of colony forming activity to those without the cytokine. These findings indicated that the bFGF is not involved in the proliferation of progenitor cells even in the presence of ST2 cells. Furthermore, bFGF inhibited osteoclast-like cell formation induced not only by 1 alpha,25(OH)2D3, but also by prostaglandin E2 and by interleukin-11. These results suggest that bFGF inhibits the common site of osteoclast-like cell formation, as induced by different mechanisms. Our data also indicated that the target cells for bFGF in inhibiting osteoclast formation are not osteoclast progenitors but stromal cells such as ST2 and osteoblastic cells, which support osteoclast development. PMID- 8707876 TI - Flow-induced modulation of the permeability of endothelial cells cultured on microcarrier beads. AB - The maintenance of endothelial barrier function is important in the regulation of fluid and solute balance between the vascular space and the surrounding tissue. Since fluid flow across endothelial cells stimulates a wide variety of endothelial responses, the effect of shear stress on barrier function was investigated. Bovine pulmonary artery endothelial cells were cultured on permeable microcarrier beads, placed in a chromatography column, and perfused. Indicator-dilution techniques were used to estimate the permeability of the cell covered beads to low molecular weight tracers (sodium fluorescein-NaFlsc; cyanocobalamin-B12) as a function of flow rate through the column. Permeability values for both tracers were significantly increased (9.3 +/- 0.6 to 19.3 +/- 1.7 for NaFlsc; 8.2 +/- 0.5 to 20.4 +/- 3.1 for B12; mean+/-SEM, x 10(-5) cm/s, P < .05) when the flow rate was increased from 0.9 ml/min to 3.2 ml/min (corresponding to average shear stresses of 4.7 and 16.8 dynes/cm2). The permeability increase occurred within minutes of the flow increase, and was reversed by decreasing the flow rate to 0.9 ml/min. In the presence of cytochalasin D, the flow-induced permeability increase was not reversible. Neither inhibition of nitric oxide synthase (with NG-monomethyl-L-arginine) nor inhibition of cyclooxygenase (with indomethacin) was capable of blocking the flow induced permeability increase. These results indicate that the rapid modulation of endothelial barrier by flow in vitro is probably not due to prostacyclin or nitric oxide. PMID- 8707877 TI - Detergent solubility defines an alternative itinerary for a subpopulation of PDGF beta receptors. AB - Current models of platelet-derived growth factor (PDGF) beta receptor itinerary are based upon the properties of receptors recovered from nonionic detergent solubilized cellular extracts. Comparing several commonly used cell extraction procedures, we have determined that up to 50% of immunoreactive PDGF beta receptors, reside in a Triton X-100 insoluble pool in a wide distribution of cultured cell lines, including Balb/c-3T3, NIH 3T3, and Swiss fibroblasts, primary murine and human fibroblasts, and primary human glial cells. Many properties of Triton insoluble receptors are distinct from the well-characterized PDGF beta receptors, including 1) delayed arrival of newly synthesized receptors into the Triton insoluble fraction, 2) prolonged half-life in the presence of PDGF, 3) increased abundance with increasing cell density, 4) inaccessibility to modification by extracellular compartment enzymes, 5) cofractionation with cytoskeletal proteins, and 6) a higher basal tyrosine phosphorylation state. PDGF stimulates accumulation of tyrosine phosphorylated PDGF beta receptors in the Triton X-100 insoluble fraction. Cell surface PDGF beta receptors modified by enzymatic desialylation redistribute to the insoluble fraction. These findings distinguish the itinerary of a large subpopulation of PDGF beta receptors from those characterized previously. Receptors in this fraction represent a long-lived tyrosine phosphorylated population that may effect responses for extended periods following ligand activation. PMID- 8707878 TI - Adaptation of chondrocytes to low oxygen tension: relationship between hypoxia and cellular metabolism. AB - In endochondral bone, the growth cartilage is the site of rapid growth. Since the vascular supply to the cartilage is limited, it is widely assumed that cells of the cartilage are hypoxic and that limitations in the oxygen supply regulate the energetic state of the maturing cells. In this report, we evaluate the effects of oxygen tension on chondrocyte energy metabolism, thiol status, and expression of transcription elements, HIF and AP-1. Imposition of an hypoxic environment on cultured chondrocytes caused a proportional increase in glucose utilization and elevated levels of lactate synthesis. Although we observed a statistical increase in the activities of phosphofructokinase, pyruvate kinase, lactate dehydrogenase, and creatine kinase after exposure to lowered oxygen concentrations, the effect was small. The cultured cells exhibited a decreased utilization of glutamine, possibly due to down regulation of mitochondrial function and inhibition of oxidative deamination. With respect to total energy generation, we noted that these cells are quite capable of maintaining the energy charge of the cell at low oxygen tensions. Indeed, no changes in the absolute quantity of adenine nucleotides or the energy charge ratio was observed. Hypoxia caused a decrease in the glutathione content of cultured chondrocytes and a concomitant rise in cell and medium cysteine levels. It is likely that the fall in cell glutathione level is due to decreased synthesis of the tripeptide under reduced oxygen stress and the limited supply of glutamate. The observed rise in cellular and medium cysteine levels probably reflects an increase in the rate of degradation of glutathione and a decrease in synthesis of the peptide. To explore how cells transduce these metabolic effects, gel retardation assays were used to study chondrocyte HIF and AP-1 binding activities. Chondrocyte nuclear preparations bound an HIF-oligonucleotide; however, at low oxygen tensions, no increase in HIF binding was observed. In addition, we found that AP-1 binding activities in chondrocytes exposed to low oxygen tensions was elevated, although the response was lower than that exhibited by fibroblasts exposed to the same range of oxygen concentrations. We compared these results to HIF and AP-1 binding by cells in the growth plate. There was strong HIF and AP-1 binding throughout the plate, but no evidence of selective binding to any one zone. The results of the study lend strong support to the view that chondrocytes are very well adapted to low oxygen tensions; thus, under hypoxic conditions, there is a high level of expression of both HIF and AP-1, and energy conservation appears to be near-maximum. PMID- 8707879 TI - Transforming growth factor-beta, osteogenin, and bone morphogenetic protein-2 inhibit intercellular communication and alter cell proliferation in MC3T3-E1 cells. AB - Intercellular communication by gap junctions has been implicated to function in the control of cell growth and differentiation in osseous tissues-processes which are regulated, in part, by peptide growth factors, including transforming growth factor-beta (TGF-beta) and the bone morphogenetic proteins (BMPs). Using the osteoblastic cell line MC3T3-E1, we tested the hypothesis that the effects of TGF beta and BMPs on cell proliferation may be correlated to changes in intercellular communication. In a series of proliferation assays, MC3T3-E1 cells were cultured in the presence of bone morphogenetic protein-2 (BMP-2) or TGF-beta for up to 48 hr. Proliferation of cells during the linear log phase (days 2 to 4) was assessed by 3H-thymidine (3H-TdR) incorporation. After times ranging from 6 to 48 hr, BMP 2 significantly inhibited uptake of 3H-TdR at doses of 50-800 ng/ml. Similarly, TGF-beta inhibited uptake of 3H-TdR at doses of 2-32 ng/ml. In a separate group of experiments, intercellular communication through gap junctions was demonstrated by cell-cell transfer of the fluorescent tracer, lucifer yellow, after microinjection. One series of experiments showed that the gap junctional intercellular communication (GJIC) of cells, incubated for 48 hr in the presence of the higher dose of osteogenin (OG) (5.0 vs. 0.5 microgram/ml) or higher dose of TGF-beta (2.0 vs. 0.2 ng/ml), was significantly inhibited compared to control. In another series of experiments, time and dose dependent effects of BMP-2 and TGF-beta on GJIC were investigated. In the time course experiments (3, 6, 12, 24, and 48 hr), TGF-beta (2.0 ng/ml) demonstrated a statistically significant effect in inhibiting GJIC as early as 6 hr, while BMP-2 (50 ng/ml) inhibited GJIC after 24 and 48 hr of treatment. The dose-dependent effects of BMP-2 and TGF-beta on cell couplings, determined at 48 hr, showed significant inhibitory effects with BMP-2 at 25 and 50 ng/ml and with TGF-beta at 2 and 4 ng/ml. The cell count results and injection study performed at 12 hr, at a fixed cell density, confirmed that the inhibitory effect was not due to differences in cell density. The 50% effective inhibitory concentrations (EC50) calculated for BMP-2 and TGF beta at 48 hr, showed no dose correlation between proliferation and GJIC, suggesting that these two events are independent occurrences. Additionally, marked morphological change was observed in the cells treated with TGF-beta. The observation may suggest that TGF-beta may have effects upon cytoskeletal elements in osseous tissues. PMID- 8707880 TI - Induction of normal and psoriatic phenotypes in submerged keratinocyte cultures. AB - Lesional psoriatic epidermis displays a number of phenotypic changes that are distinct from the differentiation program found in normal interfollicular epidermis. In psoriatic epidermis, keratinocytes are hyperproliferative and several differentiation-associated molecules are expressed that are absent in normal skin (e.g., cytokeratins (CK) 6, 16, and 17, and the epidermal proteinase inhibitor SKALP/ elafin). In addition, several molecules which are normally restricted to the stratum granulosum are strongly upregulated in the stratum spinosum (e.g., psoriasis-associated fatty acid binding protein (PA-FABP), psoriasin, involucrin, and transglutaminase). The aim of this study was to develop in vitro culture systems which (a) would allow to study the induction of normal and psoriatic differentiation pathways, and (b) would be amenable for screening of antipsoriatic drugs. Here we have investigated several models for induction of differentiation with respect to the expression of markers for the normal and psoriatic phenotype. Cell cycle parameters and expression levels of CK1, CK10, CK16, SKALP/elafin, transglutaminase, involucrin, psoriasin, and PA FABP were assessed in these models using flow cytometry, immunocytochemistry, and Northern blot analysis. We observed that induction of differentiation with fetal calf serum resembled the psoriatic phenotype (sustained hyperproliferation; high levels of CK16, SKALP/elafin, transglutaminase, and involucrin; moderate psoriasin expression), whereas differentiation induced by growth factor depletion in a confluent culture resembled the normal differentiation phenotype (low proliferative rate; high expression levels of CK1 and CK10; moderate expression of involucrin and transglutaminase; low expression levels of SKALP/elafin and CK16; absence of psoriasin). We propose that these models can be used to study expression and pharmacological modulation of selected differentiation genes and the coordinated expression of sets of genes associated with epidermal differentiation programs. PMID- 8707881 TI - Analysis of the native murine bone morphogenetic protein serine threonine kinase type I receptor (ALK-3). AB - The bone morphogenetic proteins, members of the transforming growth factor-beta cytokine family, induce the osteoblast phenotype and promote osteogenesis in the bone marrow stroma. Simultaneously, these cytokines inhibit other mesodermal differentiation pathways, such as adipogenesis and myogenesis. The receptors for the bone morphogenetic proteins belong to a family of transmembrane serine/ threonine kinase TGF beta type I and type II receptor proteins. In man, these include the activin receptor like kinase-3 (ALK-3), a type I receptor protein. We have used a polyclonal antibody to examine the expression of the native murine ALK-3 protein in murine tissues and bone morphogenetic protein-responsive cell lines. On Western blot analyses, we found that the native 85 kDa native ALK-3 protein was expressed in a number of murine tissues; protein and mRNA levels did not necessarily correlate. Two bone morphogenetic protein-responsive cell lines, BMS2 bone marrow stromal cells and C2C12 myoblasts, expressed the ALK-3 protein constitutively. Cell differentiation was accompanied by modest changes in ALK-3 protein levels. Immunoprecipitation of the ALK-3 protein cross linked to [125I] BMP-4 revealed two major receptor complexes of approximately 90 kDa and 170 kDa in size. Biotin surface-labeling experiments revealed that the 85 kDa ALK-3 protein was constitutively associated with a novel 140 kDa surface glycoprotein. Deglycosylation reduced the protein's size to 116 kDa, comparable in size to that of the recently described BMP type II receptor. These findings support the current model that BMP interacts with a pre-existing complex consisting of a type I and type II receptor protein. PMID- 8707882 TI - Low affinity purinergic receptor modulates the response of rat submandibular glands to carbachol and substance P. AB - The effect of extracellular ATP on the intracellular calcium concentration ([Ca2+]i) in rat submandibular glands was tested. The dose-response curve for ATP was biphasic with a first increase in the 1-30 microM concentration range and a further increase at concentrations higher than 100 microM. Among ATP analogs, only benzoyl-ATP stimulated the low affinity component. ATP tau S blocked this response. All the other analogs tested reproduced the high-affinity low capacity response. Magnesium and Coomassie blue selectively blocked the low affinity component. High concentrations of ATP blocked the increase of the intracellular calcium concentration [Ca2+]i in response to 100 microM carbachol. By itself, substance P (100 pM-1 microM) increased the [Ca2+]i. One mM ATP potentiated the response to concentrations of substance P higher than 10 nM. This potentiation was reversed by extracellular magnesium. Carbachol 100 microM and substance P (100 pM-1 microM) increased the release of inositol trisphosphate (IP3) from polyphosphoinositides (polyPI). Activation of the low affinity ATP receptors did not activate the polyPI-specific phospholipase C but inhibited its activation by 100 microM carbachol (-50%) and by 100 nM substance P (-60% at 1 nM substance P and -40% at 100 nM substance P). Substance P induced a strong homologous desensitization: a preincubation with 1 nM substance P nearly completely abolished the response to 1 microM substance P. When the cells were exposed to ATP before the second addition of substance P, the purinergic agonist partially restored the response to the tachykinin without totally reversing the desensitization. It is concluded that two types of purinergic receptors coexist in rat submandibular glands; a high-affinity, low capacity receptor which remains pharmacologically and functionally undefined and a low affinity site, high capacity receptor of the P2z type coupled to a non-selective cation channel. The occupancy of these low affinity sites blocks the increase of the [Ca2+]i in response to a muscarinic agonist and the activation of polyPI-specific phospholipase C by carbachol and substance P. It potentiates the effect of high concentrations of substance P on the [Ca2+]i. PMID- 8707883 TI - Amplitude-dependent modulation of brush border enzymes and proliferation by cyclic strain in human intestinal Caco-2 monolayers. AB - Little is known about the effects of repetitive deformation during peristaltic distension and contraction or repetitive villus shortening on the proliferation and differentiation of the intestinal epithelium. We sought to characterize the effects of repetitive deformation of a physiologically relevant magnitude and frequency on the proliferation and differentiation of human intestinal epithelial Caco-2 cells, a common cell culture model for intestinal epithelial biology. Human intestinal epithelial Caco-2 cells were cultured on collagen-coated membranes deformed by -20 kPa vacuum at 10 cycles/minute, producing an average 10% strain on the adherent cells. Proliferation was assessed by cell counting and 3H-thymidine incorporation. Alkaline phosphatase and dipeptidyl dipeptidase specific activity were measured in cell lysates. Since cells at the membrane periphery experience higher strain than cells in the center, the topography of brush border enzyme histochemical and immunohistochemical staining was analyzed for strain-dependence. Cyclic strain stimulated proliferation compared to static cells. Proliferation was highest in the membrane periphery where strain was maximal. Strain also modulated differentiation independently of its mitogenic effects, selectively stimulating dipeptidyl dipeptidase while inhibiting alkaline phosphatase. Strain-associated enzyme changes were also maximal in areas of greatest strain. The PKC inhibitors staurosporine and calphostin C ablated strain mitogenic effects while intracellular PKC activity was increased by strain. The strain-associated brush border enzyme changes were attenuated but not blocked by PKC inhibition. Thus, strain of a physiologically relevant frequency and magnitude promotes proliferation and modulates the differentiation of a well differentiated human intestinal epithelial cell line in an amplitude-dependent fashion. PKC may be involved in coupling strain to increased proliferation. PMID- 8707884 TI - Enhanced expression of heregulin in c-erb B-2 and c-Ha-ras transformed mouse and human mammary epithelial cells. AB - Heregulin beta 1 was found to stimulate the anchorage-dependent, serum-free growth of nontransformed human MCF-10A mammary epithelial cells. Unlike epidermal growth factor, transforming growth factor alpha, or amphiregulin, heregulin beta 1 was also able to induce the anchorage-independent growth of MCF-10A cells. In contrast, the anchorage-independent, serum-free growth of c-Ha-ras or c-erb B-2 transformed MCF-10A cells was unaffected by heregulin beta 1, whereas heregulin beta 1 was able to stimulate the anchorage-independent growth of these cells. c Ha-ras or c-erb B-2 (c-neu) transformed MCF-10A or mouse NOG-8 mammary epithelial cells express elevated levels of 2.5, 5.0, 6.5, 6.8, and 8.5 kb heregulin mRNA transcripts and/or synthesize cell-associated 25, 29, 50, and 115 kDa isoforms of heregulin. Since the MCF-10A cells and transformants also express c-erb B-3, these data suggest that endogenous heregulin might function as an autocrine growth factor for Ha-ras or erb B-2 transformed mammary epithelial cells. PMID- 8707885 TI - Identification of an osteocalcin gene promoter sequence that binds AP1. AB - Osteoblasts are differentiated cells that produce bone matrix components including the bone-specific protein osteocalcin. The osteocalcin gene promoter has become a model for understanding how genes are regulated, specifically in osteoblasts. One model for cell-specific regulation suggests that osteoblast expressed genes are regulated through common promoter sequences which bind osteoblast-specific transcriptional activators. The phenotype suppression model suggests osteoblast-specific promoters are switched off through the action of the common transcriptional activator AP1. We previously demonstrated that a short sequence element (OSCARE-2) in the osteocalcin promoter was homologous to a repressive element in the collagen type 1 (alpha 1) promoters. In this paper we use electrophoretic mobility shift (EMS) assays to examine DNA-protein interactions in the OSCARE-2 sequence. In EMS assays, OSCARE-2 binds a complex of proteins, including AP1. This supports the role of AP1 sites in contributing to the regulation of the osteocalcin promoter. Exogenous c-JUN protein bound to OSCARE-2 and increasing c-JUN incubated with nuclear extract amounts caused a progressive increase in a higher-molecular-weight complex, consistent with c-JUN involvement in protein-protein as well as DNA-protein interactions. Anti-c-FOS antibody was capable of supershifting OSCARE-2 DNA-protein complexes produced using osteoblast-like cell nuclear extracts. In addition, EMS assays of nuclear proteins from osteoblast-like cells indicated that 1,25 (OH)2D3-inducible proteins are bound to OSCARE-2. Osteocalcin promoter constructs showed that OSCARE-2 contributed to the 1,25 (OH)2D3 response, albeit in a minor way. These data support the role of AP1 protein as a regulator of osteoblast-specific gene expression during osteoblast development. PMID- 8707886 TI - Regulation of P120 mRNA levels during lymphocyte stimulation: evidence that the P120 gene shares properties with early and late genes. AB - P120 is a growth-regulated nucleolar protein, the expression of which is required for G1- to S-phase transition in lymphocytes. P120 appears to be involved in ribosomal biogenesis presumptively through its putative role as a rRNA methyltransferase. To better understand the role of P120 in cell cycle progression, we examined the regulation of the P120 gene in resting lymphocytes and in mitogen-stimulated lymphocytes as they progress from G1-phase toward S phase. P120 mRNA was detected after the immediate early gene c-fos and persisted as the cells approached S-phase. A decrease in P120 mRNA coincided with the expression of histone H3 mRNA. The level of P120 mRNA increased as cells proceeded through G1-phase, and this increase was attributed to a more than threefold increase in the P120 transcription rate and an increase in P120 mRNA stability. The P120 gene is transcribed in resting lymphocytes, although the steady-state level of P120 is small or nonexistent. P120 mRNA accumulates in resting cells in the presence of the protein synthesis inhibitor cycloheximide. Furthermore, the steady-state level of P120 mRNA increases in the presence of cycloheximide after PHA-stimulation; this level does not increase in cells not treated with this protein synthesis inhibitor. The presence of cycloheximide increases both the transcription rate of the P120 gene and the stability of P120 mRNA. These studies indicate that P120 expression is cell cycle regulated in a complex manner and that the P120 gene has properties of both early and late genes. This time ordered regulation for P120 expression may represent a necessary step for the cell cycle associated increase in ribosomal biogenesis that is required for G1-to S-phase transition. PMID- 8707887 TI - Proinflammatory agents, IL-8 and IL-10, upregulate inducible nitric oxide synthase expression and nitric oxide production in avian osteoclast-like cells. AB - Nitric oxide synthase (NOS) isoenzymes generate nitric oxide (NO), a sensitive multifunctional intercellular signal molecule. High NO levels are produced by an inducible NOS (iNOS) in activated macrophages in response to proinflammatory agents, many of which also regulate local bone metabolism. NO is a potent inhibitor of osteoclast bone resorption, whereas inhibitors of NOS promote bone resorption both in vitro and in vivo. The possibility that osteoclasts, like macrophages, express a regulated iNOS and produce NO as a potential autocrine signal following inflammatory stimulation was investigated in well-characterized avian marrow-derived osteoclast-like cells. NO production (reflected by medium nitrite levels) was markedly elevated in these cells by the proinflammatory agents lipopolysaccharide (LPS) and the synergistic action of IL-1 alpha, TNF alpha, and IFN gama. inhibitors of NOS activity (aminoguanidine, L-NAME) or iNOS induction (dexamethasone, TGF beta) reduced LPS-stimulated nitrite production. LPS also increased the NOS-associated diaphorase activity of these cells and their reactivity with anti-iNOS antibodies. RT-PCR cloning, using avian osteoclast-like cell RNA and human iNOS primers, yielded a novel 900 bp cDNA with high sequence homology (76%) to human, rat, and mouse iNOS genes. In probing osteoclast-like cell RNA with the PCR-derived iNOS cDNA, a 4.8 kb mRNA species was detected whose levels were greatly increased by LPS. Induction of iNOS mRNA by LPS, or by proinflammatory cytokines, occurred prior to the rise of medium nitrite in time course studies and was diminished by dexamethasone. Moreover, osteoclast-like cells demonstrated an upregulation of NO production and iNOS mRNA by IL-8 and IL-10, regulatory mechanism's not previously described. It is concluded that osteoclast-like cells express a novel iNOS that is upregulated by inflammatory mediators, leading to NO production. Therefore, NO may serve as both a paracrine and autocrine signal for modulating osteoclast bone resorption. PMID- 8707888 TI - Ecto-alkaline phosphatase considered as levamisole-sensitive phosphohydrolase at physiological pH range during mineralization in cultured fetal calvaria cells. AB - Alkaline phosphatase (ALP) activity expressed on the external surface of cultured fetal rat calvaria cells and its relationship with mineral deposition were investigated under pH physiological conditions. After replacement of culture medium by assay buffer and addition of p-nitrophenyl phosphate (pNPP), the rate of substrate hydrolysis catalyzed by whole cells remained constant for up to seven successive incubations of 10 min and was optimal over the pH range 7.6-8.2. It was decreased by levamisole by a 90% inhibition at 1 mM which was reversible within 10 min, dexamisole having no effect. Values of apparent Km for pNPP were close to 0.1 mM, and inhibition of pNPP hydrolysis by levamisole was uncompetitive (Ki = 45 microM). Phosphatidylinositol-specific phospholipase C (PI PLC) produced the release into the medium of a p-nitrophenyl phosphatase (pNPPase) sensitive to levamisole at pH 7.8. The released activity whose rate was constant up to 75 min represented after 15 min 60% of the value of ecto-pNPPase activity. After 75 min of PI-PLC treatment the ecto-pNPPase activity remained unchanged despite the 30% decrease in Nonidet P-40-extractable ALP activity. High levels of 45Ca incorporation into cell layers used as index of mineral deposition were decreased by levamisole in a stereospecific manner after 4 h, an effect which was reversed within 4 h after inhibitor removal, in accordance with ecto pNPPase activity variations. These results evidenced the levamisole-sensitive activity of a glycosylphosphatidylinositol-anchored pNPPase consistent with ALP acting as an ecto-enzyme whose functioning under physiological conditions was correlated to 45Ca incorporation and permit the prediction of the physiological importance of the enzyme dynamic equilibrium at the cell surface in cultured fetal calvaria cells. PMID- 8707889 TI - Expression of human mitochondrial NADP-dependent isocitrate dehydrogenase during lymphocyte activation. AB - In the process of identifying genes involved in optimization of lymphocyte activation, we have cloned the human mitochondrial NADP-dependent isocitrate dehydrogenase (mNADP-IDH) cDNA. The cDNA and its deduced amino acid (AA) sequence had a high degree of homology with those of the porcine and bovine. The heart and muscle had the highest constitutive expression of the gene. The expression of steady-state mRNA in the resting T and B lymphocytes was low but was induced after mitogen stimulation. The mRNA levels peaked around 48 h and remained elevated at 72 h. At the protein level, the mitochondrial but not cytosolic NADP IDH activity was augmented after the mitogen stimulation. There was no cell cycle dependent fluctuation of mNADP-IDH expression in synchronized Jurkat cells. In T and B cells, rapamycin (RAPA) could repress the mitogen-stimulated mNADP-IDH expression, although most of the early or late phase activation-related genes including a G-protein beta subunit-related gene H12.3 were not affected by the drug. The restricted expression of the gene in certain tissues and the activation related expression in lymphocytes suggest that this gene might be necessary for optimal functions in heart, muscle, and the activated lymphocytes. PMID- 8707890 TI - Identification and characterization of various differentiative growth plate chondrocytes from porcine by countercurrent centrifugal elutriation. AB - Countercurrent centrifugal elutriation was used to separate growth plate chondrocytes from porcine basing on their differences in sizes and densities. Eighteen fractions of cells with different sizes and densities were obtained. The mean cellular volumes increased progressively in each of successive fractions, and that increase was associated with specific phenotypic changes, such as biochemical differences in DNA synthesis, proteoglycan synthesis, and activities of alkaline phosphatase. Three distinct chondrocyte subpopulations with their unique characteristics were identified among the elutriated fractions. The resting chondrocytes were found to be small in size and quiescent. The hypertrophic chondrocytes were found to be large in size and metabolically active both in alkaline phosphatase and in proteoglycan productions. The proliferative chondrocytes exhibited a high DNA synthesis rate, and their sizes were found to be between those of the resting and hypertrophic chondrocytes. PMID- 8707891 TI - Alternative splicing of smooth muscle myosin heavy chains and its functional consequences. AB - The aim of our study was to determine the relation between alternatively spliced myosin heavy chain (MHC) isoforms and the contractility of smooth muscle. The relative amount of MHC with an alternatively spliced insert in the 5' (amino terminal) domain was determined on the protein level using a peptide-directed antibody (a25K/50K) raised against the inserted sequence (QGPSFAY). Smooth muscle MHC isoforms of both bladder and myometrium but not nonmuscle MHC reacted with a25/50K. Using a quantitative Western-blot approach the amount of 5'-inserted MHC in rat bladder was detected to be about eightfold higher than in normal rat myometrium. The amount of heavy chain with insert was found to be decreased by about 50% in the myometrium of pregnant rats. Although bladder contained significantly more 5'-inserted MHC than myometrium, apparent maximal shortening velocities (Vmax) were comparable, being 0.138 +/- 0.012 and 0.114 +/- 0.023 muscle length per second of skinned bladder and normal myometrium fibers, respectively. Phosphorylation of myosin light chain 20 induced by maximal Ca2+/calmodulin activation was the same in bladder and myometrial fibers. These results suggest that the amount of 5'-inserted MHC is not necessarily associated with contractile properties of smooth muscle. PMID- 8707892 TI - Effect of acidic phospholipids on sphingosine kinase. AB - Sphingosine-1-phosphate (SPP) is a unique sphingolipid metabolite involved in cell growth regulation and signal transduction. SPP is formed from sphingosine in cells by the action of sphingosine kinase, an enzyme whose activity can be stimulated by growth factors. Little is known of the mechanisms by which sphingosine kinase is regulated. We found that acidic phospholipids, particularly phosphatidylserine, induced a dose-dependent increase in sphingosine kinase activity due to an increase in the apparent Vmax of the enzyme. Other acidic phospholipids, such as phosphatidylinositol, phosphatidic acid, phosphatidylinositol bisphosphate, and cardiolipin stimulated sphingosine kinase activity to a lesser extent than phosphatidylserine, whereas neutral phospholipids had no effect. Diacylglycerol, a structurally similar molecule which differs from phosphatidic acid in the absence of the phosphate group, failed to induce any changes in sphingosine kinase activity. Our results suggest that the presence of negative charges on the lipid molecules is important for the potentiation of sphingosine kinase activity, but the effect does not directly correlate with the number of negative charges. These results also support the notion that the polar group confers specificity in the stimulation of sphingosine kinase by acidic glycerophospholipids. The presence of a fatty acid chain in position 2 of the glycerol backbone was not critical since lysophosphatidylserine also stimulated sphingosine kinase, although it was somewhat less potent. Dioleoylphosphatidylserine was the most potent species, including a fourfold stimulation, whereas distearoyl phosphatidylserine was completely inactive. Thus, the degree of saturation of the fatty acid chain of the phospholipids may also play a role in the activation of sphingosine kinase. PMID- 8707893 TI - Secreted MUC1 mucins lacking their cytoplasmic part and carrying sialyl-Lewis a and x epitopes from a tumor cell line and sera of colon carcinoma patients can inhibit HL-60 leukocyte adhesion to E-selectin-expressing endothelial cells. AB - A secreted MUC1 mucin from the spent medium of the colon carcinoma cell line COLO 205 carrying sialyl-Lewis a and x epitopes (H-CanAg) was purified by trichloroacetic acid precipitation and Superose 6 gel filtration. The purified H CanAg inhibited adhesion of the leukocyte cell line HL-60 to E-selectin transfected COS-1 cells or interleukin-1 beta (IL-1 beta)-activated human umbilical vein endothelial cells. Sera from two patients with advanced colon carcinoma containing high concentrations of sialyl-Lewis a and x activity inhibited HL-60 cell adhesion to E-selectin-expressing COS-1 cells and IL-1 beta activated endothelial cells. After affinity column absorption of the sialyl-Lewis a activity, the sera also lost most of their sialyl-Lewis x activity and at the same time their adhesion inhibitory effect. A large part of the sialyl-Lewis a/x activity in the two patients was found in fractions containing mucins having a MUC1 apoprotein, as shown by its size, and reactivity with the two anti-MUC1 apoprotein monoclonal antibodies, Ma552 and HMFG-2. The cell-adhesion inhibitory effect of the purified sialyl-Lewis a-carrying MUC1 mucin fraction from the sera of the two patients was stronger than that of smaller sized sialyl-Lewis a carrying mucin-type glycoproteins also found in the patient sera. The MUC1 mucin fraction secreted by the COLO 205 cells and from the two sera were all shown to lack their C-terminal portion, in contrast to the MUC1 mucin from cells. It is hypothesized that sialyl-Lewis a- and/or x-containing mucins, especially MUC1, secreted by tumors can interact with E-selectin on endothelial cells and thus inhibit leukocyte adhesion. PMID- 8707894 TI - LFA-1/ICAM-3 mediates neutrophil homotypic aggregation under fluid shear stress. AB - We found that human neutrophils undergo homotypic aggregation by loading the physiological range of fluid shear stress (12-30 dynes/cm2). Under the fluid shear stress, an increase of intracellular Ca2+ concentration of neutrophils was observed. This increase of intracellular Ca2+ concentration was caused by Ca2+ influx, and the blockage of the flux by NiCl2 suppressed the neutrophil homotypic aggregation. Furthermore, this neutrophil aggregation under fluid shear stress was completely inhibited by pretreatment with antibody against LFA-1 or ICAM-3. These results suggested that NiCl2-sensitive Ca2+ channel played an important role in LFA-1/ICAM-3-mediated neutrophil homotypic aggregation under fluid shear stress. PMID- 8707895 TI - Analysis of human breast cancer nuclear proteins binding to the promoter elements of the c-myc gene. AB - The expression of the c-myc gene is essential for the proliferation of both hormone-dependent and -independent human breast cancer cells. The regulation of c myc gene expression in MCF-7 (hormone-dependent, estrogen-receptor (ER)-positive) and MDA MB 231 (hormone-independent, ER-negative) human breast cancer cells differs, with the c-myc gene of MCF-7 but not MDA MB 231 cells being regulated at the transcriptional level by estrogen. We have shown previously that the DNAase I hypersensitive (DH) sites in the c-myc chromatin of hormone-dependent and independent human breast cancer cells were similar, with the exception of DH site II2. DH site II2, which maps near the P0 promoter, was less sensitive in hormone dependent than in hormone-independent cells. As DH sites generally indicate the presence of sequence-specific DNA-binding proteins, we undertook a study to identify the nuclear proteins isolated from MCF-7 and MDA MB 231 cells that bound to the P0 and P2 promoter regions of the c-myc gene in vitro. The studies presented here provide evidence that Sp1 and/or Sp1-like proteins bind to the P0 and P2 promoter regions of the c-myc gene of MCF-7 and MDA MB 231 cells. Furthermore, evidence is presented for the presence of several previously unidentified sequence-specific DNA-binding proteins binding to these promoters. The DNA-binding activities of these latter proteins differed in the nuclear extracts of the MCF-7 and MDA MB 231 human breast cancer cells. PMID- 8707896 TI - Basic fibroblast growth factor selectively regulates ornithine decarboxylase gene expression in malignant H-ras transformed cells. AB - Cell growth regulation by fibroblast growth factors (FGFs) is highly complex. The present study demonstrates a novel link between alterations in bFGF regulation during malignant conversion and the expression of ornithine decarboxylase, a key rate-limiting and regulatory activity in the biosynthesis of polyamines. H-ras transformed mouse 10T 1/2 cell lines exhibiting increasing malignant potential were investigated for possible bFGF-mediated changes in ornithine decarboxylase gene expression. Selective induction of ornithine decarboxylase gene expression was observed, since, in contrast to nontransformed 10T 1/2 cells and cells capable of only benign tumor formation, H-ras transformed metastatic cells exhibited marked elevations in ornithine decarboxylase message levels. Evidence for regulation of ornithine decarboxylase gene expression by bFGF at both transcription and posttranscription was found. Actinomycin D pretreatment of malignant cells prior to bFGF exposure inhibited the increase in ornithine decarboxylase message. Furthermore, striking differences in the rates of ornithine decarboxylase message decay were observed when cells treated with bFGF were compared to untreated control cells, with the half-life of ornithine decarboxylase mRNA increasing from 2.4 h in untreated cells to 12.5 h in cells exposed to bFGF. Evidence was also obtained for a cycloheximide-sensitive regulator of ornithine decarboxylase gene expression whose effect, in combination with bFGF, resulted in a further augmentation of ornithine decarboxylase gene expression. Furthermore, evidence is presented to suggest a possible role for G protein-coupled events in the bFGF-mediated regulation of ornithine decarboxylase gene expression. The bFGF regulation of ornithine decarboxylase expression in H ras transformed malignant cells appeared to occur independent of protein kinase C mediated events. These results show that bFGF can modulate ornithine decarboxylase gene expression in malignant H-ras transformed cells and further suggests a mechanism of growth factor stimulation of malignant cells wherein early alterations in the regulatory control of ornithine decarboxylase gene expression are critical. PMID- 8707898 TI - Clinical practice as scholarship. PMID- 8707897 TI - Endogenously inhibited protein kinase C in transgenic Drosophila embryonic neuroblasts down regulates the outgrowth of type I and II processes of cultured mature neurons. AB - Embryonic neurons were cultured from transgenic Drosophila melanogaster expressing a highly specific pseudosubstrate inhibitor of protein kinase C (PKC). Flies homozygous for this transgene, which is under the control of the yeast UAS promoter, were crossed to flies homozygous for the yeast heat shock inducible transcription factor GAL 4. Following heat shock, the progeny express the pseudosubstrate inhibitor at high levels. This strategy, which has the advantage of avoiding the non-specific effects of drugs, was used to study the role of PKC in process growth of cultured, differentiating neuroblasts. An external gold particle labeling procedure using a cell surface antigen expressed by mature neurons and processes was used to visualize neuronal processes directly in the scanning electron microscope. We observed that cell cultures expressing a low concentration of the pseudosubstrate inhibitor showed a significant decrease in the number of type I and II processes as compared to control cultures, while the proportions of neuroblasts, ganglion mother cells (GMCs), and mature neurons in the clusters were little affected. PMID- 8707899 TI - Critical thinking in clinical learning: a holistic perspective. AB - A dynamic clinical setting affords multiple opportunities for learning. The article describes undergraduate and graduate teaching/learning strategies that facilitate the development of critical thinking and holistic intervention competencies. Insights regarding outcome evaluation of these competencies and the use of data for curriculum changes are presented. PMID- 8707901 TI - Innovative strategies for promoting clinical scholarship: a holistic approach. AB - The article discusses a holistic approach to clinical scholarship by examining dimensions of scholarship and patterns of knowing. Innovative clinical teaching strategies in relation to these dimensions of scholarship and patterns of knowing are identified. Experiential learning, content mapping, and portable patient problem packs are explored. Knowledge building and creativity in the process of educating students are imperative. Nurse educators are challenged to develop new paradigms for nursing education that create a dynamic learning environment and result in enhanced clinical scholarship. PMID- 8707902 TI - Assessment design issues for evaluating critical thinking in nursing. AB - The assessment of graduating students' critical thinking skills and habits of mind challenges and rewards those who approach the task from a critical thinking perspective. The article identifies and discusses issues in the design of authentic assessments of critical thinking as an educational outcome predictive of competent professional judgment in nursing practice. Authentic assessment implies a multiple methods design that addresses the diverse contexts within which judgments must be made by professional nurses. Most important, it implies a concern for validity and reliability of measurement, selection of appropriate data points, and attention to a number of logistic and practical concerns. PMID- 8707900 TI - Metacognition: state-of-the-art learning theory implications for clinical nursing education. AB - Clinical nursing education represents one of the most challenging aspects of the faculty role because nursing educators are being required to teach crucial aspects of comprehensive clinical practice to students in limited time periods and in increasingly demanding, high-acuity affiliation sites. State-of-the-art research in metacognition provides a stimulating array of instructional strategies that can assist in this process and provide an impetus for further cognitive inquiry in nursing. The article analyzes metacognition, explores its historical roots, delineates its relationship to memory theory, and describes a range of metacognitive strategies that are useful to faculty and students in nursing. PMID- 8707903 TI - Sharing the quest for knowledge through interdisciplinary research. AB - Faced with increasingly complex health care issues and diminishing resources for research, nurse scientists must collaborate with each other and with scientists from other disciplines in the search for innovative and cost-effective health care solutions. The article traces nursing's 20-year struggle to become a credible partner in interdisciplinary research and highlights advantages and challenges characteristic of such collaboration. The authors describe their respective experiences as members of interdisciplinary research teams, one in a private medical practice affiliated with a for-profit hospital and the other in an academic setting. Recommendations that nurture successful collaborations are proposed. PMID- 8707904 TI - The lived experience of nursing students in caring for suffering individuals. AB - Most nursing school curricula address the pathologic nature of illness, the nursing care of clients who are ill, the prevention of illness, and the promotion of health. There seems to be minimal discussion, however, both in nursing curricula and in textbooks, of the alleviation of suffering or of how nurses know that someone is suffering. The article describes a phenomenologic analysis of 12 nursing students' lived experiences of caring for individuals who were suffering. The voices of the students as they chronicled their experiences are exemplars of clinical scholarship as described in the literature. PMID- 8707905 TI - The scholarship of a practice discipline. AB - The article proposes that all nurses, as members of a discipline that is embedded in practice, share in the obligation to take part, in different capacities, in the generation of nursing scholarship. A model of scholarly inquiry that connects knowledge development and professional values to gather wisdom that benefits the health and well-being of society is advanced. Opportunity for all nurses to engage in a dialogue to create ownership of their scholarship is pivotal to its revisioning. PMID- 8707906 TI - Helping say goodbye: merging clinical scholarship with community service. AB - Clinical scholarship is elusive but involves observation, comparison of what is and what might be, applicability and analysis of appropriate theory-based practice, and passion for the process. The article examines characteristics of clinical scholarship in a clinical practice with a community service mission: facilitation of a support group for parents experiencing perinatal loss. The conceptual framework for this practice arena, Swanson-Kauffman's caring concepts and Yalom's curative factors of group participation, is explored. Practical information about initiating and maintaining an environment of clinical scholarship in a support group practice is provided, with discussion of the appropriateness of support group involvement for this population. PMID- 8707907 TI - Utilizing all opportunities in clinical practice. AB - Clinical practice provides an abundance of avenues through which nurses may enhance both clinical judgment and scholarly activities. Creative ways to stimulate opportunities for this knowledge exist. The article describes the approaches utilized by clinical nurse specialists in two teaching hospitals. A cardiovascular nursing unit and a neurologic nursing unit served as settings for description. Advanced practice nurses provided motivation. PMID- 8707908 TI - Application of genetic methodologies to the study of behaviour. PMID- 8707909 TI - Annotation: the impact of puberty. PMID- 8707910 TI - Mental retardation: genetic findings, clinical implications and research agenda. AB - The most important genetic advances in the field of mental retardation include the discovery of the novel genetic mechanism responsible for the Fragile X syndrome, and the imprinting involved in the Prader-Willi and Angelman syndromes, but there have also been advances in our understanding of the pathogenesis of Down syndrome and phenylketonuria. Genetic defects (both single gene Mendelizing disorders and cytogenetic abnormalities) are involved in a substantial proportion of cases of mild as well as severe mental retardation, indicating that the previous equating of severe mental retardation with pathology, and of mild retardation with normal variation, is a misleading over-simplication. Within the group in which no pathological cause can be detected, behaviour genetic studies indicate that genetic influences are important, but that their interplay with environmental factors, which are also important, is at present poorly understood. Research into the joint action of genetic and environmental influences in this group will be an important research area in the future. PMID- 8707911 TI - Adolescent resiliency to family adversity. AB - The factors associated with adolescent resiliency to childhood adversity were examined in a birth cohort of 940 New Zealand adolescents studied to the age of 16 years. Resilient teenagers were defined by: (a) high exposure to family adversity during childhood and (b) an absence of a wide range of externalising problems during adolescence including substance abuse, juvenile offending and school problems. Resilient teenagers were characterised by significantly higher IQ (p < .001), lower novelty seeking (p < .01) and lower affiliations with delinquent peers (p < .005) with these factors acting accumulatively to influence the probability of resilience to externalising problems. PMID- 8707912 TI - Subtypes of incarcerated delinquents constructed via cluster analysis. AB - The MMPI and Jesness Inventory were used to construct groupings of juvenile delinquents homogeneous with respect to personality characteristics. Subjects were 191 incarcerated juvenile delinquents, aged 12 to 19. All participants had been adjudicated on one or more serious offenses. Five subtypes of delinquents were identified. They included an alienated, unsocialized group, an insecure anxious group, a group reporting limited difficulties with peers but alienated from adults, a group with no apparent emotional disturbance, and a group with multiple elevations. Significant differences between clusters with regard to self reported explosiveness, suicidality, gang activities, substance abuse, history of sexual abuse, and delusions and hallucinations were identified. Discriminant function analysis was then used to support the generalizability of the classification system to an independent sample. Eighty per cent of the second sample were accurately classified. The subtypes identified are compared to delinquent classification systems developed previously. PMID- 8707913 TI - Distractibility in hyperactive and conduct-disordered children. AB - Hyperactive children are often said to be inattentive and distractible. However, the results from a number of experimental studies are equivocal. To examine this discrepancy, a Chinese version of the Stroop Test was devised. Four groups of subjects recruited from a community sample of 1479 Chinese boys living in Hong Kong took part in the investigation. These were: (1) a pure hyperactive (HA) group; (2) a mixed hyperactive/conduct-disordered (HA + CD) group; (3) a pure conduct-disordered (CD) group; and (4) a normal (N) control group. The results revealed that HA children were more markedly affected by the introduction of distracting stimuli. This distractibility was probably a function of both stimulus potency and the random order in which stimuli were presented. The specificity of a greater distractibility to HA children indicates its diagnostic value with regard to hyperactivity. The failure to find a similar deficit in HA + CD children raises questions about the clinical identity of this mixed diagnostic group. PMID- 8707914 TI - Test-retest reliability for false-belief tasks. AB - Despite notable variations in children's rate of success on theory of mind tasks and the presumed theoretical implications drawn from a child's success or failure on such tasks, there have been no studies of the test-retest reliability of children's performance on these tasks. Twenty-three children (mean age 49.6 months, SD 8.6) watched three videotaped stories illustrating a false-belief situation: the standard experimenter narrated false-belief task, a minor variant replacing the narration of the story with a dialogue among the characters, and a third version involving a humorous situation. The time elapsed between test and retest was 2-3 weeks and the order of presentation was counterbalanced. Results corroborated previous findings of a developmental trend in the understanding of false-belief questions but, despite a general improvement in children's comprehension of the stories, the test-retest reliability for the false belief questions was poor. Although changes recorded between test-retest sessions frequently occurred in the direction of children answering correctly questions they had previously failed, a subset of children incorrectly answered questions they had initially passed. These findings underscore the need for validation assessments of techniques for studying children's developing theories of mind. PMID- 8707915 TI - Family functioning predictors of adjustment in children with newly diagnosed cancer: a prospective analysis. AB - Theoretically-driven investigations on the potentially modifiable predictors of individual differences among newly-diagnosed pediatric cancer patients may facilitate the identification of children at risk for adjustment problems. Within a risk and resistance conceptual model, family functioning was investigated concurrently and prospectively as a predictor of adjustment in newly-diagnosed pediatric cancer patients at Time 1 (within 1 month after diagnosis), Time 2 (6 months postdiagnosis), and Time 3 (9 months postdiagnosis). The family relationship dimensions of cohesion and expressiveness most consistently predicted the psychological and social adjustment of children with newly diagnosed cancer over a 9-month period after initial diagnosis. These findings are discussed in terms of the treatment implications for enhancing child adjustment to newly-diagnosed cancer and biomedical treatment. PMID- 8707916 TI - What's real in children's fantasy play?: fantasy play across the transition to becoming a sibling. AB - This study examined children's fantasy enactments during the critical life transition of becoming a sibling to determine the degree to which the thematic content of fantasy play reflects children's realistic concerns. Thirty 3-5-year old children played with their best friend at two pre- and three post-birth intervals. Mothers kept diary records of children's concerns, worries, and events they were excited about during 3-day periods surrounding the five play sessions. Raters then evaluated the degree to which the thematic content of the fantasy play was consistent with their concerns as reported by mothers. Overall, little convergence was apparent between the themes of fantasy play and children's perceived concerns. However, a modest relationship was found between acting out positive concerns in play and sibling relationship quality. Although these results lend limited support for the notion that children's concerns are reflected in the content of their spontaneous play, they do suggest that 'playing it out' is beneficial for children. PMID- 8707917 TI - The validity of mixed disorders of conduct and emotions in children and adolescents: a research note. AB - In a study of differential validity, three diagnostic groups based on ICD-10 criteria were compared, namely children with emotional disorders, children with conduct disorders, and children with mixed disorders of conduct and emotions. The patients were taken from a consecutive series of child and adolescent psychiatric service attenders from 1988 to 1992 and represented the total entire subgroups of patients with the respective diagnoses. Based on standardized item sheets that were filled out by the clinicians who were responsible for the respective cases, the groups were compared with regard to age, sex ratio, family background, premorbid behavioral abnormalities, abnormal psychosocial situations, and treatment. It was found that children with mixed disorders of conduct and emotions share many characteristics with pure conduct disordered children. It is concluded that the former group is a subgroup of the latter, and there is little evidence in this study supporting the validity of a separate category of mixed disorders of conduct and emotions. PMID- 8707918 TI - [Chromosomal translocation and molecular characteristics in hematological malignancies]. PMID- 8707919 TI - [Suppressive effect of Gossypol, Gossypolone and serum from Gossypol treated rats on steroidogenesis in cultured porcine granulosa cells]. AB - Gossypol (GP), a male antifertility agent, and Gossypolone (GN), a major metabolite of GP, have been shown to have a suppressive effect on steroidogenesis in bovine luteal cells. We examined the effect of GP, GN and serum obtained from rats given GP (GP-serum) on steroidogenesis in porcine granulosa cells (PGC). To obtain GP-serum, 50-60 day old Copenhagen rats received GP (12.5 mg/kg B.W./day, sc) for 14 days. PGC were incubated with GP, GN and GP-serum at different doses for 24 hours. GP and GN significantly suppressed Progesterone (P4) and Estradiol (E2) release in a dose-dependent manner, respectively. GP-serum also significantly suppressed P4 release in a dose-dependent manner. The magnitude of suppression of P4 obtained by GP-serum could not be explained by the amount of GP remaining in the serum. Therefore, the inhibition of P4 release by GP-serum could not be solely attributed to GP. PMID- 8707920 TI - [Flow cytometric measurement of DCFH-oxidation of neutrophils in peripheral whole blood, isolated leukocytes and cerebrospinal fluid cells]. AB - Using 2',7'-dichlorofluorescein diacetate (DCFH-DA) as an indicator for the intracellular formation of H2O2 and free radicals, we measured DCFH-oxidation (DCF fluorescence) of neutrophils by flow cytometry. The DCF fluorescence intensity of unstimulated neutrophils in whole blood was the same as the autofluorescence of neutrophils. However, that of unstimulated isolated neutrophils was higher than autofluorescence. The DCF fluorescence of isolated neutrophils was higher than that of neutrophils in whole blood with or without stimuli, and was decreased to the intensity of neutrophils in whole blood by the addition of plasma or erythrocytes. The DCF fluorescence of neutrophils in whole blood indicates the oxidative state of neutrophils in vivo. The determination of DCFH-oxidation of neutrophils in isolated leukocytes is also necessary to evaluate precisely the oxidative state of neutrophils. In cerebrospinal fluid (CSF) cells from patients with meningitis, the DCF fluorescence intensity of unstimulated neutrophils was higher than the autofluorescence of CSF neutrophils. The DCF fluorescence of CSF neutrophils stimulated with PMA showed increased intensity over that of unstimulated CSF neutrophils. The DCF fluorescence of CSF neutrophils, both unstimulated and stimulated with PMA, were decreased with the addition of plasma but not with the addition of CSF. PMID- 8707921 TI - [Investigation of gastric cancers detected at a medical check-up center]. AB - Most of the gastric cancers investigated in this study were detected during mass screening at a Medical Check-up Center. The research period was 5 years, from 1990 to 1994. The total number of patients undergoing initial gastric examinations was 300,658. Four point eight percent of these needed detailed examinations, and 77.2% actually underwent detailed examinations. Over the 5 years, the number of gastric cancers detected was 253. The rate of detection of these cancers was almost unchanged every year, with the average rate of change being 0.11%. The rate of detected gastric cancers was investigated according to age and sex. No cancers were found below age 29, and the rate gradually rose over age 30. Over age 50, the rate in males was twice that in females. The rate of early gastric cancers was 66.4% of all reported cancers. As regards location of 253 cases, 16.5% were in the C-area, 45.8% in the M-area, 36.2% in the A-area; 19.2% were in the greater curvature, 33.9% were in the lesser curvature, 17.3% in the anterior wall, and 26.1% in the posterior wall. The sizes of the lesions were as follows: 11.5% were below 1.0 cm, 29.6% were from 1.1 cm to 2.0 cm, 46.6% were from 2.1 cm to 5.0 cm, and 12.3% were over 5.1 cm. It was considered that indirect X-rays were slightly inferior to direct X-rays in detecting early gastric cancer. The X-ray positionings in which cancers were detected were: 75.7% by supine double contrast and 48.6% by compression in 144 cases of early cancer. On the other hand, the rate were 75.6% by supine double contrast, and 51.2% by compression in 82 cases of advanced cancer. A similar tendency was found in one shot X-ray positioning which revealed cancers. The rate of gastric cancer notdetected by X-ray pictures was 5.9% of the 253 cases. The size of the cancers notdetected by X-ray was within 2.0 cm in all cases. 58.7% of the 155 early cancer patients and 48.2% of the 83 advanced cancer patients had examinations the previous year. Therefore, it is clear that some cases of advanced cancer were not detected in the mass gastric screenings. Endoscopically, 7 cases of gastric cancer were diagnosed correctly by means of repeated biopsies which were needed 3 or 4 times over 3 to 18 months owing to pseudonegative findings on the first bioptic examination. Consequently, it is necessary to make naked eye diagnosis by endoscopic examination. Six cases of death from gastric cancer were certified within one year after normal diagnosis during the mass screening. Three cases were Borr. 4, 2 cases Borr. 3, and 1 case was Borr 2. A retrospective investigation of X-ray pictures showed that it would have been difficult to identify the lesions in these cases. The above results show that the accuracy of examinations and diagnosis must be raised in mass gastric screenings, but it is doubtful whether relying on the present methods of screening will lead to a marked improvement. For the purpose of increasing the effectiveness of mass gastric screening, we would emphasize the necessity of the following new tests; serum pepsinogen measurement, reinvestigation of patient's ages, shortening the intervals between examinations in high risk groups, using direct X-rays rather than indirect X-rays, and intermitted endoscopic examinations. PMID- 8707922 TI - [Analysis of the 5'-flanking regions of adult type beta-globin genes in rats]. AB - In order to elucidate the molecular mechanism of the tissue- and stage-specific expression of globin genes, the possible regulatory role of the 5'-flanking region of the rat beta-globin genes was investigated by determining the nucleotide sequence and measuring the CAT transfection activity of a series of deletion mutants. Results obtained in the present study are summarized as follows: 1. Transcription initiation sites of the rat II beta-, III beta, and O beta-globin genes were determined to be 52 base pairs (bp) 5'-upstream of the translation initiation codon (ATG), in each gene by primer extension analysis. 2. The nucleotide sequences of 5'-flanking regions of each gene (II beta: -3819/+51, III beta: -716/+51, 0 beta: -727/+51) were determined. Dot matrix search of the individual sequences revealed the occurrence of about 300 bp of alternating purine and pyrimidine repeats at a region extending from the nucleotide positions -757 to -1049 5' to the transcription initiation site of the II beta-globin gene. 3. CAT transfection assay using constructs with deletions in 5'-flanking regions indicated that a negative regulatory effect occurs in the region between -757 and -1049 of the II beta-globin gene, corresponding to that containing the purine/pyrimidine repeats mentioned above. These results strongly suggest that the presently identified purine/pyrimidine stretch may act as a repressor. 4. Dot matrix analysis between the 5'-flanking nucleotide sequence of the rat II beta globin gene and those of adult type beta-globin genes of other species showed that the repeat sequence existing in the 5'-flanking region of the rat gene is very homologous to that found in a similar region of the mouse beta-major globin gene, which has been reported to contain a possible repressor element. PMID- 8707923 TI - [Endocrine surgery. Series 6: Alteration of RET proto-oncogene and clinical implication in medullary thyroid carcinoma. Genetic screening of family members at risk for inherited medullary thyroid carcinoma]. PMID- 8707924 TI - [Treatment of acute heart failure up-to-date]. PMID- 8707926 TI - Soluble complement receptor type 1 in serum and cerebrospinal fluid of patients with Guillain-Barre syndrome and multiple sclerosis. AB - Activation of complement is critically involved in inflammatory reactions in both Guillain-Barre syndrome (GBS) and multiple sclerosis (MS). Soluble human complement receptor 1 (sCR1) blocks complement activation by both classical and alternative pathways. We studied serum and cerebrospinal fluid (CSF) concentrations of sCR1 in 23 patients with GBS, 27 patients with MS and 30 controls. No significant differences were found between patients and controls. Transient liver affection probably caused high serum sCR1 levels in two patients with GBS. The serum and CSF sCR1 levels were not correlated to the disease activity of GBS and MS, nor to the relapsing-remitting or chronic-progressive forms of MS. In GBS the CSF sCR1 levels correlated with the CSF total protein concentrations (r = 0.9, P < 0.01), suggesting that sCR1 leaks from serum into CSF via a damaged blood-nerve barrier. The serum sCR1 levels in GBS were slightly higher than in MS (P < 0.05). Whether this reflects changes in the release or consumption of sCR1 in these patients is at present unknown. PMID- 8707925 TI - Lateralization of cutaneous inflammatory responses in patients with unilateral paresis after poliomyelitis. AB - Unilateral paresis remaining after poliomyelitis may affect the expression of inflammatory diseases by lateralization of the disease manifestations. The purpose of this study was to assess the impact of the unilateral paresis after poliomyelitis on lateralization of neurogenic inflammation and immune responsiveness. The delayed-type hypersensitivity (DTH) reaction to tuberculin was used as an in vivo measure of antigen-specific T lymphocyte reactivity. Assessment of axon reflex vasodilatation was simultaneously employed to test for neurogenic inflammation. Fourteen of the 16 polio patients displayed a positive DTH reaction to tuberculin. All but two showed weaker DTH reaction on the paretic compared to the contralateral-side (P = 0.001). Magnitude of electrically evoked axon reflexes significantly correlated to asymmetries of DTH responses. We conclude that damage of lower motor neuron leads to ipsilateral down-regulation of T cell-mediated cutaneous inflammation. This lateralization of DTH responses is related to deficiencies in motor and sympathetic innervation of the paretic extremity. PMID- 8707927 TI - beta-Amyloid induces increased release of interleukin-1 beta from lipopolysaccharide-activated human monocytes. AB - Previous reports have demonstrated that IL-1 is elevated in the Alzheimer's disease brain. We propose that beta-amyloid (A beta) in senile plaques triggers microglial interleukin-1(IL-1) release. Since microglia and monocytes have similar lineage and functions, the human monocyte cell line, THP-1, was used to determine whether A beta peptides can stimulate release of IL-1 beta. THP-1 cells were grown in culture with LPS and incubated with various A beta peptides (0.5-10 microM). IL-1 released into the medium was measured using either an IL-1 beta ELISA or an IL-1 bioassay. Treatment of activated THP-1 cells with A beta 25-35, fibrillar A beta 1-40, or A beta 1-42 significantly elevated IL-1 beta release. A beta 25-35 with a scrambled sequence or non-fibrillar A beta 1-40 did not significantly change IL-1 beta release from activated THP-1 cells. The A beta 25 35- and fibrillar A beta 1-40 induced IL-1 beta release was dose-dependent. IL-1 released following treatment with A beta 25-35 and measured using an IL-1 bioassay gave similar results. The present report provides evidence that A beta is capable of elevating release of functional IL-1 beta, a potent pro inflammatory cytokine, from macrophages/microglia and provides support that a chronic local inflammatory response is an ongoing phenomenon within and surrounding senile plaques. PMID- 8707928 TI - Sex differences in susceptibility to viral infection of the central nervous system. AB - We have characterized striking differences in recovery of male and female BALB/c and BALB/c-H-2dm2 (dm2) mice from an experimental neurotropic viral infection of the central nervous system (CNS). Following intranasal inoculation of vesicular stomatitis virus (VSV), assays of tissue homogenates from female mice produced lower viral titers. There was also a significant reduction in the spread of virus from the rostral to caudal end of the brain in female mice. Enhanced recovery by female mice of both strains in response to this viral insult correlates with increased levels of Nitric Oxide Synthase (NOS) types I, II, and III expression, an increased prevalence of reactive astrocytes, earlier and enhanced levels of expression of Major Histocompatibility Complex (MHC) class II molecules on astrocytes, endothelial and microglial cells, and increased T cell infiltration in the female BALB/c mouse. Taken together, these findings document sexual dimorphism in CNS immunity, and may provide an understanding of some of the mechanisms underlying many sex-biased diseases. PMID- 8707929 TI - Comparison of saturable transport and extracellular pathways in the passage of interleukin-1 alpha across the blood-brain barrier. AB - Blood-borne cytokines enter the brain by transport across the blood-brain barrier (BBB) or by leakage through extracellular pathways at sites, such as circumventricular organs (CVOs), without a BBB. We used radioactively labeled albumin (T-Alb) to differentiate the relative contribution of transport and extracellular pathways to the passage of interleukin-1 alpha ([125I]IL-1 alpha) across the BBB. The major mechanism of entry for [125I]IL-1 alpha after intravenous (i.v.) injection was a saturable transport system with extracellular pathways accounting for only a small fraction of entry into brain. CVOs concentrated blood-borne [125I]IL-1 alpha in a saturable manner to a much greater extent than did the cerebral cortex and cerebellum, but accounted for less that 5% of total brain uptake. After intracerebroventricular (i.c.v.) injection, [125I]IL-1 alpha and T-Alb were concentrated in the CVOs, especially the median eminence, although CVOs contained less that 1% of the substances injected. Distribution after i.c.v. injection was largely due to diffusion and leakage through extracellular pathways. We conclude that after i.c.v. injection, leakage across extracellular pathways accounts for the small but concentrated amount of [125I]IL-1 alpha found in CVOs. After i.v. injection, transport across the BBB accounts for the majority of [125I]IL-1 alpha in brain. PMID- 8707930 TI - Recognition of neurokinin 1 receptor (NK1-R): an antibody to a peptide sequence from the third extracellular region binds to brain NK1-R. AB - Substance P (SP) can produce cytokine-like responses by astrocytes and mononuclear cells. In an effort to identify neurokinin-1-receptors (NK1-R), an antibody to NK1-R was generated by using a linear peptide sequence from the deduced third extracellular region (ECR) corresponding to the seven transmembrane rat brain NK1-R. The ECR-3 peptide was coupled to keyhole-limpet hemocyanin and the antisera produced in rabbits was purified by binding to a peptide-affinity matrix. The specificity for the anti-peptide antibody was shown by its reactivity to the ECR-3 peptide by ELISA. The anti-ECR-3 peptide antibody could detect, by Western blot analysis of SDS-PAGE-separated rat brain membranes, a single band with an apparent molecular weight (MW) of 53-54 kDa. An affinity matrix made from the anti-ECR-3 antibody was used to isolate NK1-R from rat brain membranes which exhibited two products on SDS-PAGE with apparent MW of 54 and 44 kDa. The C6 astrocytes were shown to express NK1-R as determined by [125I]Bolten-Hunter SP binding to intact cells with a Kd = 0.32 nM. These C6 cells did not co-express either NK2-R or NK3-R when analyzed at the mRNA level. The anti-ECR-3 peptide antibody could inhibit [125I]Bolten-Hunter SP binding to intact C6 astrocytes and CHO cells expressing NK1-R by greater than 95% when compared to normal rabbit IgG which failed to inhibit radiolabeled SP binding. Thus, an antibody which recognizes surface determinants to the NK1-R could be generated upon immunization with an NK1-R peptide. PMID- 8707931 TI - A novel enzyme-linked immunosorbent assay for determination of synaptophysin as compared with other quantification procedures. AB - A two-sided enzyme-linked immunosorbent assay (ELISA) has been established for reliable, specific and sensitive determination of synaptophysin (SYN), an intrinsic membrane protein of the small synaptic vesicles. This ELISA used a highly specific monoclonal antibody (SY 38) as capture reagent and a specific SYN antiserum in combination with a secondary peroxidase-conjugated antibody for detection. Calibration was carried out with immunoaffinity-purified SYN from porcine cortex. The sensitivity was found to be improved substantially when the ELISA was compared with previously used dot-immunobinding assays. This ELISA allowed rapid and reliable determination of SYN from detergent lysed homogenates, partially and highly purified preparations of rat, porcine and human brain. SYN was determined in highly purified small synaptic vesicles, and it was calculated to be 5.8% of total detergent solubilized protein. PMID- 8707932 TI - Acute electrical stimulation of lateral hypothalamus increases natural killer cell activity in rats. AB - Natural killer cell (NK) activity in WKA and SD rats was found to be significantly higher following electrical stimulation of the lateral hypothalamus (LH) compared to sham operated. There was no such difference between sham operated rats and those receiving electrical stimulation in the frontal cortex as a control. Operations were performed under sodium pentobarbital anesthetic, and NK activity against YAC-1 target cells was measured 20 h later using 51Cr release assay. The LH area stimulated is a potent reward center and that stimulation of this point increased NK activity opens the possibility that pleasure might play a role in cellular immunity. PMID- 8707933 TI - Beta-adrenergic receptor regulation of macrophage-derived tumor necrosis factor alpha production from rats with experimental arthritis. AB - Prostaglandin E2 (PGE2) and beta-adrenergic agonists can suppress lipopolysaccharide-induced tumor necrosis factor-alpha (TNF) production from elicited macrophages. We assessed the responsiveness of rat peritoneal macrophages to PGE2 and the beta-adrenergic agonist isoproterenol during immunologically-mediated arthritis. We assessed macrophage sensitivity to these mediators from resident macrophages and macrophages elicited with either streptococcal cell wall or complete Freund's adjuvant. Peritoneal macrophages were obtained from female Lewis rats that were (1) injected with complete Freund's adjuvant and non-arthritic (CFA); (2) injected with streptococcal cell wall and arthritic (ART); (3) injected with streptococcal cell wall and non reactive (NON) and (4) non-elicited resident macrophages (RES). When challenged with graded concentrations of lipopolysaccharide (0.1 to 10,000 ng/ml), macrophages obtained from each group of rats released TNF in a concentration dependent manner, with macrophages from arthritic rats (ART) producing the greatest amount of TNF (p < 0.001). While PGE2 suppressed lipopolysaccharide (100 ng/ml) stimulated TNF production in a concentration-dependent manner in all groups, the greatest sensitivity to PGE2 was observed with macrophages obtained from rats which received streptococcal cell wall when compared to both complete Freund's adjuvant-elicited and resident macrophages (p < 0.05). The beta adrenergic agonist isoproterenol also inhibited lipopolysaccharide-stimulated TNF production from macrophages in all groups. In addition, the specific beta 2 adrenergic antagonist, ICI 118.551, shifted isoproterenol concentration-effect curves to the right (p < 0.01). Minimal responsiveness to isoproterenol was observed with resident peritoneal macrophages. Maximum isoproterenol-induced inhibition of TNF production was observed with complete Freund's adjuvant elicited macrophages, and significantly less in macrophages of streptococcal cell wall-injected rats. Of particular interest, macrophages obtained from streptococcal cell wall-injected rats, which became arthritic, were significantly less sensitive to isoproterenol than those which did not develop arthritis (p < 0.02). In addition, these changes in sensitivity were not reflected by changes in the sensitivity of both CFA and ART groups to dibutyryl cAMP. The present study demonstrates a shift in the balance between inhibitory mediator responses in rats inoculated with one of two different adjuvants. These investigations support the role of PGE2 and a neurotransmitter as immunomodulating compounds which may effectively maintain an inflammatory lesion such as arthritis. PMID- 8707934 TI - Inaugural meeting of the Brain-Immune Network Group (BING). PMID- 8707935 TI - The use of flash visual evoked potentials in the early diagnosis of suspected optic nerve lesions due to craniofacial trauma. AB - Craniofacial trauma encroaching on the orbital apex and optic canal can result in direct or indirect optic nerve lesions, leading to visual impairment or blindness. Early diagnosis of a visual loss and immediate therapy are generally considered crucial for a successful restoration of vision in indirect trauma. However, in comatose or sedated patients the assessment of optic nerve function by testing pupillary reactivity may be severely compromised or impossible because of tensely swollen eyelids, conjunctival oedema, concussion of the ciliary muscle or pharmacological effects. In the event that clinical ophthalmic examination, computer tomography or nuclear magnetic resonance scanning fail to clarify the state of the optic nerve, visual evoked potentials (VEPs) to flash stimulation appear to provide reliable information on function within the visual pathway. On this basis, treatment with corticosteroids and/or surgical decompression can be rapidly initiated. Our results in a preliminary patient series confirm the value of acutely monitored VEPs as an objective test of optic nerve function in cases of suspected optic nerve injury immediately after admission to the emergency care unit. The imaging techniques usually applied may be complemented by VEPs to show the functional significance of structural abnormalities found in the vicinity of the optic nerve. PMID- 8707936 TI - Complications following arthroscopy of the temporomandibular joint: analysis covering a 10-year period (451 arthroscopies). AB - 451 arthroscopies of the temporomandibular joint (TMJ) have been performed on 373 patients during 10 years of clinical experience. The complication rate was low (1.77%). Most complications were transient, consisting mainly of temporary deficits of the 5th and 7th cranial nerves. No patient required arthrotomy because of complications, but one patient had to undergo angiography and embolization of a traumatic aneurysm of the superficial temporal artery secondary to arthroscopy. PMID- 8707937 TI - Blood supply to the oral and maxillofacial tissues following radiation therapy: a prospective ultrasonographic study. AB - In this prospective clinical study of the early radiation effects on blood supply to the mouth and face, 44 patients (31 men-13 women, mean age 63.4 yrs) with oral tumors underwent surgery and adjuvant postoperative radiotherapy with conventional fractionation (mean dose at the neck approximately equal to 50 Gy). Blood flow parameters 1 cm below the bifurcation of the common carotid artery (mean velocity time averaged, lumen diameter, resistivity index-RI) as well as perivascular reaction, were recorded on Color Doppler Imaging video tapes in a series of five consecutive examinations up to six months postirradiation. There were no statistically significant changes shown between the initial and follow-up examinations for any of the parameters investigated. There was no difference in blood flow between the ipsilateral (operated-irradiated) and contralateral side of the neck. Results did not seem to correlate with known vascular disease risk factors such as sex, arterial pressure, cholesterol levels, smoking and diabetes. After the effect of age was controlled, flow measurements remained statistically stable. Radiation dosage did not appear to influence carotid flow parameters. Perivascular reaction had the highest peak immediately postirradiation but regressed with time. This study suggests that therapeutic radiation of the neck at this dose level may not have important effects on the maxillofacial region blood supply for approximately eight months postoperatively; however, these patients should be closely evaluated for symptoms or signs of carotid artery lesions on a long-term basis. PMID- 8707938 TI - Histological changes in vessels used for microvascular reconstruction in the head and neck. AB - Recipient vessels from the head and neck region were histologically examined in 30 patients who had undergone extensive tumor resection necessitating microvascular tissue reconstruction. Past illnesses considered to be risk factors included hypertension, myocardial disease, lung disease, cirrhosis, diabetes, nephritis, as well as extensive nicotine and alcohol abuse. Blood vessels chosen for microsurgical anastomosis were exclusively examined histologically in this study. Patients undergoing microvascular surgery demonstrated vessel abnormalities in 93%. The frequency of dysplasia was higher in the arteries than in the veins (73% and 26%, respectively). Marked thickening of the blood vessel wall and severe exfoliation of the endothelial cells were observed in most arteries. Fibrodysplasia and exfoliated endothelial cells were more frequently observed in the recipient arteries than the graft arteries. Only slight thickening of the vessel wall and mild fibrodysplasia were seen in the veins. Two graft failures were correlated to technical errors rather than pre-existing vessel lesions. This study revealed that most patients undergoing microsurgery in the head and neck region demonstrate pre-existing damage in vessels, which generally hinders anastomosis. Although the study tried to identify fully the interrelationships between the extent of dysplasia, past medical history, preoperative therapy, risk factors and factors that cause free-graft failure, pre existing changes in the recipient and graft vessels may cause technical difficulties and must be regarded as additional factors contributing to graft failure. PMID- 8707939 TI - Bone grafting in the mixed and permanent dentition in cleft lip and palatepatients: long-term results and the role of the surgeon's experience. AB - This long-term study was a follow-up of 46 patients with complete unilateral cleft lip and palate treated with secondary or late secondary bone grafting. Surgeons with different degrees of experience in this procedure had operated on these patients. Morphology of the clefts prior to the grafting was assessed with regard to width of cleft, stage of eruption of permanent canine, presence or absence of permanent lateral incisor and position of permanent lateral incisor and permanent canine in the dental arch. Outcome of surgery was evaluated with regard to the amount of bleeding, flap dehiscence, alveolar bone height, and space closure in the previous cleft area. Dehiscence rate was 23% (11) and total failure rate 4% (2). Alveolar bony height in 38 patients (81%) was more than 75% of the normal alveolar bone height; in 6 cases (13%) between 50-75% bone height and in 3 subjects (6%) surgery had given less than 50% of the normal bone height. Cleft space was closed by orthodontic means in 49% of patients. Best results were obtained when bone grafting was performed prior to canine eruption. Furthermore, significantly better results regarding alveolar bone height were obtained by the more experienced surgeons. PMID- 8707940 TI - Case report: hypoglossia-hypodactylia syndrome. AB - A case of hypoglossia-hypodactylia syndrome is described. The infant was characterized by micrognathia, accompanied by a defect of the gingiva, and hypoplasia of the tongue. There was defect in the right arm below the elbow. The right foot manifested syndactyly of the great, second and third toes and osseous fusion of the first and second metatarsi and proximal phalanges. In the left foot brachydactyly of the great and fifth toes, syndactyly of the second, third and fourth toes, hypoplasia of the first and second metatarsi and defect in the first phalanx were evident. PMID- 8707941 TI - Twenty-five years experience in the treatment of trigeminal neuralgia. Comparison of three different operative procedures in forty-nine patients. AB - A series of 49 patients with trigeminal neuralgia (TN) were treated by three different surgical procedures: (1) peripheral ablative procedures in 10 patients; (2) percutaneous rhizotomy in 17 patients and (3) intracranial rhizotomy (IR) in 22 patients. On the basis of surgical treatment, the concept that neurovascular compression is a mechanical factor in the aetiology of TN was supported in 14 to 18 patients who underwent posterior fossa exploration. The results support the conclusion that retromastoid craniectomy with IR is the procedure of choice for the majority of patients with TN. PMID- 8707942 TI - Computerized axiographic evaluation of condylar movements in cases with fractures of the condylar process: a follow up over 19 years. AB - The treatment of fractures of the mandibular condylar process(es) can be conservative or surgical. However, in many cases, a definitive judgment on the functional long-term outcome of the chosen therapy can only be given many years after the intervention. As a result, it is clearly useful to be able to review the effects of treatment undertaken at least 15 years previously. Even so, only two such studies have been identified. Therefore, the aim of the present study is to evaluate the current functional capacity of the mandibular condyles of 20 subjects who had had an uni- or bilateral fracture of the condylar process, on average 19 years ago. The individuals, who all had been treated conservatively, were compared with a control group of healthy volunteers matched for gender and age. Besides measuring maximum mouth opening (MMO), computerized axiographies in the sagittal plane were made for each condyle during MMO, maximum protrusion and maximum mediotrusion. In spite of the high degree of inter-individual variability in both groups, the data showed that the amount of condylar movement was in most cases greater in the control group. In general, it appears that the risk of developing functional problems after conservative treatment is highest in the case of a fracture of the condylar process accompanied by condylar luxation, rather than by a condylar dislocation or without it. PMID- 8707943 TI - Experimental analysis of internal rigid fixation osteosynthesis performed with titanium bone screw and plate systems. AB - The authors report the results of an experimental analysis performed on titanium miniplates and screws in order to gain a better understanding of dynamic forces in internal rigid fixation. Ten segments of bovine scapula were prepared. Osteotomies were carried out along the minor axis, following which five were fixed with four hole straight miniplates and the other five with six hole double Y miniplates. Each sample was fastened in a special clamp adapted to a tension test machine and shearing force was applied. Force versus time was recorded and the 50 bone fragments were examined by a pathologist. On the basis of the test results, two simple computer models were developed. No significant difference was evident between the mechanical and computed tests. The most critical sections were located near the hole proximal to the osteotomy and the microscopic findings confirmed this. On the basis of the experimental results, the authors propose a new plate design in which the area subject to most stress, proximal to the bone section, would be of miniplate thickness, the distal aspect being thinner as in a microplate. It is suggested that this design would provide sufficient stability and a high degree of anatomical adjustment of the system. PMID- 8707944 TI - A biomechanical comparison between original and used titanium miniplates. AB - Four dogs were chosen for this study. An experimental fracture was made in the mandibular symphysis region of each dog. Two miniplates and eight screws were applied, and removed three and six months postoperatively. In this study, the surface and mechanical characteristics of titanium miniplates were evaluated. Study materials consisted of four groups. In each of the groups, there were six miniplates. The groups are as follows: Group A: Original plates (not used) as a control (i.e. used as a reference in comparisons) Group B: Original plates bent as in operation but not left in tissue Group C: Plates removed at third month postoperatively Group D: Plates removed at sixth month postoperatively. After evaluation of all groups, there was no difference in the microstructure of the surfaces but some differences in mechanical characteristics were apparent. PMID- 8707945 TI - Extracellular matrix and the kidney. PMID- 8707946 TI - Intracerebral haemorrhage. PMID- 8707947 TI - Significance of endotoxin in lethal synergy between bacteria associated with sudden infant death syndrome: follow up study. AB - AIM: To investigate the role of endotoxin in synergy between bacterial toxins associated with sudden infant death syndrome (SIDS). METHODS: Extracellular toxins of 13 isolates of Staphylococcus from SIDS victims and matched healthy infants were tested for lethal toxicity in chick embryos with and without standard endotoxin (used at 1.00 ng/embryo). Endotoxin and toxins from staphylococci were used at dilutions with negligible lethality. RESULTS: Simultaneous injection of non-lethal levels of endotoxin and toxins from 11 of the 13 staphylococcal isolates tested produced lethal toxicity that was 111 to 613% greater than expected by an additive effect alone. This was highly significant and occurred even in the absence of staphylococcal enterotoxins or toxic shock syndrome toxin-1. CONCLUSION: Endotoxin enhancement of staphylococcal toxin lethality could be partly responsible for the clinical outcome in SIDS. PMID- 8707948 TI - A rapid and highly specific technique to detect hepatitis C RNA in frozen sections of liver. AB - AIMS: To develop a polymerase chain reaction (PCR) based technique that would permit the rapid and highly specific detection of hepatitis C virus (HCV) RNA extracted from frozen liver tissue. METHODS: Samples of liver tissue from 18 patients undergoing orthotopic liver transplantation were studied. Nine patients were HCV positive. Total RNA was extracted from between one and 10 sections, 10 microns thick, from each tissue sample. HCV RNA was amplified by (1) conventional, multistep reverse transcription PCR (RT-PCR) and by (2) combined, single step RT-PCR using coupled oligonucleotide primers based on the sequence of the 5' untranslated region of the viral genome. Positive results were confirmed by dot blot analysis using a digoxigenin labelled oligoprobe (Alx 89). RESULTS: HCV RNA was detected in the nine HCV positive patients by both conventional and combined RT-PCR. HCV RNA was not detected in the HCV negative patients. As little as 500 ng total RNA was needed as the template to yield detectable amounts of amplified cDNA. The digoxigenin labelled oligoprobe hybridised with HCV RNA positive specimens only. CONCLUSIONS: The combined, single step RT-PCR is a rapid and sensitive technique for detecting HCV RNA in frozen liver tissue. PMID- 8707949 TI - Accuracy of serology for the diagnosis of Helicobacter pylori infection--a comparison of eight kits. AB - AIMS: To determine the accuracy of eight commercially available kits for the serological diagnosis of Helicobacter pylori infection, and hence whether a serology service could be introduced to reduce endoscopy workload. METHODS: Eighty four patients newly presenting to their general practitioners with dyspepsia were recruited. Gold standard diagnosis of H pylori infection was obtained both by a histological examination of gastroduodenal biopsy specimens and by the 14C-urea breath test (UBT). The performance of six quantitative and two qualitative enzyme linked immunosorbent assays for H pylori IgG, used according to the manufacturers' instructions, with serum samples obtained during the endoscopy visit, were compared. RESULTS: The study population had a median age of 45 years, and the prevalence of H pylori infection was 35%. With one exception, where the patient had received a course of anti-H pylori treatment between endoscopy and UBT, there was 100% concordance in the results of the two gold standard techniques. Discordant serology results were more common in patients aged > 50 years (42% of the total) than in younger patients (21%), and this was most noticeable in uninfected patients. The sensitivity of the kits was good (90-100%), but specificity was more variable (76-96%), and the rate of equivocal results was unacceptably high in some cases (0-12%). The overall accuracy of the kits ranged from 83 to 98%. Two kits in particular performed well (Pylori-Elisa II, Bio-Whitaker and Premier, Launch; qualitative) with 98% and 100% accuracy, respectively. CONCLUSIONS: In a symptomatic population with a prevalence of H pylori infection of 35%, particularly in patients aged < 50 years, some but not all serology kits may be used as a highly accurate and inexpensive alternative to the gold standard techniques. PMID- 8707950 TI - Histology of chronic gastritis with and without duodenitis in patients with Helicobacter pylori infection. AB - AIM: To compare the histological characteristics of Helicobacter pylori positive chronic gastritis in patients with and without associated duodenitis. METHODS: Gastric mucosal biopsy specimens were obtained from patients undergoing endoscopy for dyspepsia. Severity of gastritis and density of H pylori infection were graded according to the Sydney system. RESULTS: Of the 69 patients studied, 15 had normal histology, 22 had chronic gastritis only (77.3% H pylori positive), 21 had duodenitis (90.5% H pylori positive), and 11 had other diagnoses. In the H pylori positive patients, the median gastritis score was higher in the duodenitis group (6, range 3-9) than in the chronic gastritis only group (5, range 2-8), because of greater neutrophil activity scores in patients with duodenitis (median score 2 v 1). There were no differences in the density of H pylori infection, inflammation, atrophy, or intestinal metaplasia between patients with chronic gastritis only and those with duodenitis. CONCLUSIONS: These results suggest that H pylori positive patients with duodenitis have a more severe form of gastritis than those without associated duodenal inflammation. This is because of increased neutrophil activity, which seems to be independent of the density of H pylori infection. PMID- 8707951 TI - Monocyte activation in patients with non-seminomatous germ cell tumour of the testis before and after tumour eradication. AB - AIMS: To investigate the kinetics of normalisation of monocyte oxidative activity following tumour eradication. METHODS: Whole blood lucigenin enhanced chemiluminescence was studied in patients with non-seminomatous germ cell tumours. Group 1 comprised 14 patients who had been "cured" of their cancer (the term "cured" as used in this report denotes a relapse free period of at least three years). Group 2 comprised 15 patients who were followed from diagnosis to up to two years after the start of treatment. RESULTS: Lucigenin enhanced chemiluminescence of whole blood in the "cured" patients was similar to that of controls and lower than that in patients who had not yet received chemotherapy (group 2). After treatment, chemiluminescence decreased slowly and did not normalise until 18 months after the start of treatment. Tumour necrosis factor alpha (TNF alpha) concentrations were normal in "cured" patients but were raised in those who had not yet received treatment. TNF alpha was normalised 12 months after start of treatment. Alpha-fetoprotein concentrations were raised in most patients but normalised rapidly after tumour eradication. CONCLUSIONS: The activity of blood monocytes, as measured by whole blood lucigenin enhanced chemiluminescence, is increased in cancer. This activity may be a consequence of the presence of tumour cells. Immunocompetent cells remain active for over a year after eradication of the tumour. PMID- 8707952 TI - Chromosome 11q in sporadic colorectal carcinoma: patterns of allele loss and their significance for tumorigenesis. AB - AIMS: To analyse the frequency of loss of heterozygosity (allele loss, LOH) in a large sample of colorectal carcinomas using highly informative markers along chromosome 11q. METHODS: One hundred paired samples of colorectal cancer and normal tissue were genotyped at six microsatellite markers on chromosome 11q (cen D11S1313-D11S901-DRD2/NCAM-D11S29- D11S968-tel). The high levels of heterozygosity at these markers allow allele loss to be determined in about 80% of cases at any one locus. The frequency of replication errors (RERs, microsatellite instability) has also been determined. RESULTS: LOH was found at frequencies of 25% and 29% at the distal D11S968 (11qter) and D11S29 (11q23.3) loci, slightly above the accepted baseline of 0-20%. Allele loss at NCAM, DRD2, D11S901, and D11S1313 was not raised above baseline levels. The probable genetic mechanism of allele loss--chromosomal non-disjunction, mitotic recombination, deletion, or gene conversion--seemed to vary between tumours and no consistent mechanism of mutation was found. Microsatellite instability was found in 23 (23%) tumours. No associations were found between LOH and clinical data (patient sex, age at presentation, tumour site, and Duke's stage). CONCLUSIONS: Although gene(s) on 11q may have a role in the development of a minority of colorectal carcinomas, this study provides evidence against the general importance of allele loss on chromosome 11q in the pathogenesis of colorectal cancer. The results also have implications for the importance of 11q in other cancers: it seems less likely that a single tumour supressor gene at this location promotes the growth of all types of tumour when lost. Rather, one or more genes with tissue specific effects may be involved. PMID- 8707953 TI - Splenic macrophages in preterm infants: a necropsy study. AB - AIMS: To study the frequency and nature of histiocytes in the splenic red pulp of infants who died following complicated immaturity/prematurity. METHODS: Twenty four preterm/immature infants were investigated. Frozen sections of formalin fixed splenic tissue were stained with Oil Red O. Paraffin wax sections from the same tissue were conventionally stained with haematoxylin and eosin. Immunohistochemistry was carried out for a number of macrophage markers. The administration of Intralipid was compared with the presence and extent of tissue macrophages. RESULTS: The spleens of 10 infants showed varying degrees of Oil Red O positivity ranging from mild to strong. In all these cases varying numbers of macrophages were confirmed in the splenic parenchyma in ordinary sections. The immunomarkers indicated that the histiocytes belonged to the macrophage phagocytic system. Of the 10 cases with splenic macrophages all had received Intralipid. Of those not receiving Intralipid none showed splenic macrophages. Seven had received Intralipid but did not have splenic macrophages; they had either only received small amounts of Intralipid or Intralipid was discontinued before death. CONCLUSIONS: Splenic macrophages are common at necropsy in immature/preterm infants. The macrophages are most lucidly demonstrated using Oil Red O staining in frozen sections. There is a strong association between the presence of splenic macrophages and Intralipid administration. PMID- 8707954 TI - Immunohistochemical analysis of bcl-2 expression in transitional cell carcinoma of the bladder. AB - AIMS: To evaluate the expression of bcl-2 in transitional cell carcinoma (TCC) of the bladder; to compare bcl-2 expression with clinicopathological findings, p53 immunoreactivity, proliferating cell nuclear antigen (PCNA) expression, 2c deviation index (2cDI), 5c exceeding rate (5cER), and the mean nuclear area (MNA). METHODS: Cystectomy specimens from 77 patients with untreated, non metastatic TCC of the bladder were studied. Expression of bcl-2, p53 and PCNA was detected immunohistochemically using the following monoclonal antibodies: bcl 2/124, DO-7 and PC10, respectively. Nuclear DNA content was analysed using static cytometry. RESULTS: Bcl-2 was expressed in 19 (24.7%) of 77 TCCs and in 74 (96.1%) of 77 normal samples of transitional epithelium (taken from normal tissue adjacent to the tumour in each case). In all cases, bcl-2 immunoreactivity was more intense in normal transitional epithelium than in TCC. In normal transitional epitehlium and superficial TCC bcl-2 immunoreactivity was observed at the basal layer, and not at the invasive front. Bcl-2 immunoreactivity was invesely correlated with histological grade and p53 immunoreactivity, and was not correlated with the pT category, disease progression, PCNA expression, 2cDI, 5cER, and the MNA. No significant correlation was found between bcl-2 expression and overall survival. CONCLUSIONS: Bcl-2 expression in TCC of the bladder seems to be associated with a less aggressive phenotype and does not play an important role in tumour progression. PMID- 8707955 TI - Thymidine phosphorylase expression in Kaposi sarcoma. AB - AIMS: To examine the immunohistochemical distribution of thymidine phosphorylase (TP) in all clinicopathological subtypes of Kaposi sarcoma. METHODS: Thirty two biopsy specimens of Kaposi sarcoma (29 patients) were studied. Six of these patients represented classic, six endemic, eight HIV associated, seven post immunosuppression/transplant related, and two unclassified variants of Kaposi sarcoma. The average age was 49 years (range 22-83 years) and the male: female ratio 24:5. Four samples of angiosarcoma and one of spindle cell haemangio endothelioma were stained in parallel. All specimens were fixed in formalin, embedded in paraffin wax and processed routinely. Immunohistochemistry was carried out using an antibody directed against CD31 (JC70) and the monoclonal antibody P-GF.44C against TP. RESULTS: All biopsy specimens showed immunoexpression for TP. The spindle cell component stained more strongly than newly formed endothelium lined vessels and normal, resident vessels at a distance from the lesions. CONCLUSIONS: The strong immunoexpression of TP suggests up regulation of TP and a role for TP in angiogensis in Kaposi sarcoma. The mechanism for the up-regulation of TP remains unknown, but viral infections may trigger it. The differential staining of the various cell components of Kaposi sarcoma also suggest that TP either plays a role in the differentiation and maturation of Kaposi sarcoma or is a reflection of such changes. PMID- 8707956 TI - Necropsy study of the association between sudden cardiac death, cardiac isoenzymes and contraction band necrosis. AB - AIMS: To assess whether a quantitative analysis of myocardial contraction bands could aid the postmortem identification of early myocardial infarction, especially if used in conjunction with cardiac isoenzyme activities. METHODS: Sixty four coroner's necropsies were grouped by gross and histological findings into 26 cases of definite non-cardiac death, 12 cases of definite myocardial infarction and 26 cases in which there was occlusive coronary artery atheroma, but no demonstrable evidence of infarction. Using multiple sections of left ventricular myocardium stained with Heidenhain's iron haematoxylin, the number of myocardial cells containing contraction bands per unit area was quantified. The results were analysed statistically using logistic regression, and were then compared and combined with results from the statistical analysis of postmortem cardiac isoenzymes that had recently been undertaken on the same cases. RESULTS: The number of cells containing contraction bands per unit area was higher in cases of definite myocardial infarction compared with those of non-cardiac deaths. In addition, cases of occlusive coronary artery atheroma only could be identified, indicating the presence of early myocardial infarction. The accuracy of this identification could be improved by combining these results with the results from the statistical analysis of postmortem cardiac isoenzymes. CONCLUSION: The quantitative assessment of myocardial contraction band necrosis can provide useful additional information in cases of sudden death, where myocardial infarction is suspected but not identified on routine histological staining. The value of the information obtained is increased when used in conjunction with the postmortem measurement of cardiac isoenzyme activities. PMID- 8707957 TI - The pathology of cornea in Tangier disease (familial high density lipoprotein deficiency). AB - AIMS: To clarify the underlying causes of corneal opacification in Tangier disease. METHODS: Both corneas were removed at death from a 62 year old man with Tangier disease, and were examined by direct and transmission electron microscopy, histochemistry, biochemical analysis by thin-layer and gas-liquid chromatography after extraction, and by differential scanning calorimetry. RESULTS: Membranous inclusions in the stroma were seen on transmission electron microscopy. Direct analysis confirmed enrichment with phospholipids and cholesterol, with acyl patterns and proportions as ester broadly similar to those of normal cornea. Tangier cornea showed major thermotropic phase transitions in the range 28-37 degrees C, peak 30-33 degrees C, extending above profiles of normal clear cornea and without the complexity of those seen with cornea with heavy arcus involvement. CONCLUSIONS: Lipid accumulation underlies corneal opacification in Tangier disease. The excess material is mainly phospholipid and cholesterol esters. As at other sites which are below body core temperature, notably tonsil, accumulation may be enhanced by local impaired mobilisation of material as the phase transitions of the excess lipid present extend above ambient corneal temperatures. PMID- 8707958 TI - Comparison of the HbH inclusion test and a PCR test in routine screening for alpha thalassaemia in Hong Kong. AB - AIM: To compare the haemoglobin (Hb) H inclusion test with a polymerase chain reaction (PCR) test in routine screening for alpha thalassaemia. METHODS: Ninety nine peripheral blood samples from Chinese patients with mean corpuscular volume below 80 fl were screened for alpha thalassaemia using the HbH inclusion test and by PCR utilising primers bridging the common deletion breakpoint of the South East Asian (--SEA/) deletion. RESULTS: The HbH inclusion test was positive in 78 (79%) patients, 73 (93.7%) of whom carried the (--SEA/) deletion on analysis of their DNA by PCR, as did one patient with a negative HbH inclusion test. CONCLUSIONS: These results suggest that in areas with a high prevalence of the (- SEA/) deletion, such as Hong Kong, the HbH inclusion test can be replaced by PCR as the investigation of choice in screening for alpha thalassaemia. PMID- 8707959 TI - Multicentre evaluation of the Thrombotest International Sensitivity Index used with a steel ball coagulometer. AB - AIM: To compare the International Sensitivity Index (ISI) of the Thrombotest reagent used with a steel ball coagulometer (KC) to the ISI of the same reagent used with the manual (tilt tube) technique. METHODS: The study was carried out by eight laboratories using their own KC instrument and method of testing. All laboratories used the same batch of Thrombotest to determine the clotting times of fresh blood samples from 20 local healthy volunteers and 60 patients on long term oral anticoagulant therapy. KC clotting times were plotted against manual clotting times on double logarithmic scales. Orthogonal regression lines were calculated to assess the ISI. RESULTS: In two laboratories the ISI of the KC method was lower than that of the manual method; these differences, however, were 2% or less. In the other laboratories no clinically important differences were observed between ISI values obtained. However, the clotting times determined with the KC methods were shorter than the manual values. CONCLUSIONS: The ISI of Thrombotest determined with the KC methods was very similar to the manual value. Therefore, use of the ISI value supplied by the manufacturer without adjustment is justified. The mean normal prothrombin time, however, must be determined locally. PMID- 8707960 TI - Cellular distribution of androgen receptors in the liver. AB - In order to determine the cellular distribution of androgen receptors (AR) in normal liver and to examine whether phenotypic changes occur in a variety of non neoplastic liver diseases, cryostat sections of explanted livers removed from 52 consecutive patients undergoing orthotopic transplantation were immunostained using an anti-androgen receptor monoclonal antibody. In histologically normal liver, AR was immunolocalised to the nuclei of hepatocytes. The proportion of positive hepatocytes varied from about 50% to greater than 90%. Staining, of variable intensity, was restricted to parenchymal cells with no evidence of zonal heterogeneity with respect to labelling intensity. In tissue from patients with biliary cirrhoses and in some cases of alcoholic cirrhosis, labelling for AR was observed in areas of ductular metaplasia but not in areas of "typical" ductular reaction (ductular proliferation). Otherwise, no consistent abnormalities in immunolabelling were seen in any of the diseased livers. PMID- 8707961 TI - Alpha-fetoprotein production by a hepatoid adenocarcinoma of the uterus. AB - A case of a 62 year old Japanese woman with an endometrial adenocarcinoma producing alpha-fetoprotein (AFP) is described. Microscopically, the tumour was composed of a major medullary portion and a minor tubular adenocarcinoma which had invaded the myometrium, the myometrial lymphatics and blood vessels. Neoplastic cells in the medullary portion were polygonal with glycogen-rich cytoplasm. Vascular permeation by neoplastic cells was prominent. Extensive hepatoma-like features were observed. The tumour cells lacked features suggestive of a diagnosis of embryonal carcinoma or endodermal sinus tumour. The production of AFP by the tumour cells was demonstrated immunohistochemically using the PAP technique. Only two cases of AFP producing endometrial adenocarcinomas have been reported previously. PMID- 8707962 TI - Anti-GM1 antibodies in polyneuropathies of unknown origin. AB - This study was undertaken to determine whether anti-GM1 titres are raised in polyneuropathies of unknown origin and whether determination of these titres is useful for diagnosing these conditions. The study population comprised 20 controls (aged 36-88 years), 12 patients with polyneuropathies of known origin (aged 31-81 years) and 15 patients with polyneuropathies of unknown origin (aged 40-77 years). Antibody levels were measured using a commercial GM1 enzyme linked immunosorbent assay kit (Buehlmann Laboratories). Mean anti-GM1 IgG and IgM antibody titres were not raised in patients with polyneuropathies of unknown origin. Anti-GM1 IgG antibody titres were raised in one and GM1 IgM antibody titres in none of the patients with polyneuropathies of unknown origin. In conclusion, GM1 antibody levels are rarely raised in polyneuropathies of unknown origin and probably play a minor role in the pathogenesis of these conditions. PMID- 8707963 TI - Quantification of HIV DNA in the brain by PCR: differences between fresh frozen and formalin fixed tissue. AB - HIV-1 DNA extracted from frozen and formalin fixed brain tissue can be detected using PCR. This work has been extended by amplifying, using semiquantitative PCR, HIV DNA extracted from frontal lobe tissue of 16 patients with AIDS (eight positive and eight negative for p24 antigen). DNA was amplified using HIV-1 pol gene digoxigenin labelled primers and detected by chemiluminescence and densitometry. Cloned standards were amplified in parallel for quantification. HIV DNA levels detected in frozen tissue showed a correlation with p24 positivity and the severity of the histological diagnosis. This correlation was less clear in the formalin fixed material. PMID- 8707964 TI - Myelofibrosis presenting as chronic cholecystitis. AB - A 61 year old man presented with abdominal pain typical of chronic cholecystitis of one month's duration. Pallor was noted on examination and investigation uncovered myelofibrosis and a small gallstone. Cholecystectomy relieved the pain and pathological examination of the gall bladder showed widespread myeloid metaplasia. This is the first reported case of myelofibrosis presenting as chronic cholecystitis. PMID- 8707965 TI - Review of clinical activity by microbiologists. AB - A data form was devised and used to collect information on clinical cases involving a microbiologist. From the results a relational database management system was created. Of a total of 280 interventions, 137 (49%) were proactive, and in 118 (86%) of these cases the advice given was accepted. The majority of the patients in these cases showed subsequent improvement. Of all the interventions, the given advice was acted upon in 235 (84%), in 22 (8%) it was not and for the remainder this information was not available. This study was a simple method of gaining information on the clinical involvement of the microbiology department of a large city hospital. It provides a reference point from which further research and audit can be based. PMID- 8707966 TI - Granulomatous disease in common variable immunodeficiency: effect on immunoglobulin replacement therapy and response to steroids and splenectomy. AB - A 40 year old white woman with common variable immunodeficiency of four years duration presented with rapidly increasing splenomegaly. Despite high dose, weekly intravenous immunoglobulin, it was impossible to raise the trough serum IgG concentration to within the normal range. While waiting for a diagnostic splenectomy, low dose corticosteroids were started, leading to a decrease in the size of the spleen and an increase in the trough IgG concentration. Both spleen and liver showed non-caseating granulomas. Following splenectomy, the corticosteroids were tailed off and the trough IgG was maintained well into the normal range on a reduced, fortnightly dose of intravenous immunoglobulin and a low dose of oral corticosteroid. PMID- 8707967 TI - Failure to kill Yersinia enterocolitica by plasma diluted to the concentration found in red cell units. AB - The possibility that the use of additive solutions for red cell storage might impair the ability of plasma to kill Yersinia enterocolitica was investigated by studying killing of Y enterocolitica by neat and diluted plasma. The ability of neat citrated plasma to kill complement sensitive organisms was lost at around 26%, the dilution typically found in red cell units. These results should be considered in the light of evidence that killing in plasma is important in the protection of donated blood against growth of Y enterocolitica, and the observation that the increase in frequency of transfusion reactions caused by Y enterocolitica coincided with the widespread introduction of additive solutions. Taken together, these points support the suggestion that the introduction of additive solutions may have precipitated the problem of growth of Y enterocolitica in stored blood. PMID- 8707968 TI - Coronary artery dissection. PMID- 8707969 TI - Neomycin blood agar as a selective medium for vancomycin resistant Enterococcus faecium. PMID- 8707970 TI - Effects of interleukins on the proliferation and survival of chronic lymphocytic leukaemia cells. PMID- 8707971 TI - Effectiveness of a prescale gel on subgingival calculus. AB - A prescale gel product, designed to facilitate the removal of calculus, has recently been introduced and marketed to the dental profession. The purpose of this study was to evaluate the effectiveness of this gel on the removal of subgingival calculus. 10 patients, each with 5 periodontally diseased teeth scheduled for extraction, participated in this in vivo/in vitro study. 4 teeth per patient were randomly assigned using a 2-by-2 block design and treated in vivo with either active or placebo gel, with or without scaling, prior to extraction. To assess possible overexposure to the product, 10 selected teeth from the sample were treated with active gel for an extended exposure time. Standardized scaling was performed on a 4x4 mm treated root area in vitro on groups as assigned. Quantification of residual calculus was determined by one examiner blind to treatment group assignment using SEM photomicrograph montages and the Java image analysis computer system. Repeated measures ANOVA showed no statistically significant treatment effect for gel (p > 0.05) in the scaled and no-scaled groups. The 5th group exposed to the prescale gel for an extended time was evaluated descriptively for root surface morphological changes with no noticeable effect. Based on the results of this investigation, treatment of subgingival calculus with prescale gel offers no advantage for calculus removal over scaling alone. The findings suggest no significant clinical impact of product use. PMID- 8707972 TI - Efficacy of systemically administered acetylsalicylic acid plus scaling on periodontal health and elastase-alpha 1-proteinase inhibitor in gingival crevicular fluid. AB - The purpose of this proof of principle trial was to assess whether conventional periodontal therapy and systemically administrated acetylsalicylic acid (ASA) are functionally synergistic when combined in the treatment of periodontitis. A total of 30 patients with untreated moderate to severe adult periodontitis were enrolled into the study and were given placebo q.i.d. between the baseline and 6 week examination, and acetylsalicylic acid (ASA) 500 mg q.i.d. between the 6-week and 12-week examinations. In addition, they received supra- and subgingival scaling in 1 quadrant after baseline examination and in 2 further randomly selected quadrants after the 6-week examination. The study design resulted in the following 4 therapies: (1) scaling plus ASA 500 mg q.i.d.; (2) scaling plus placebo q.i.d.; (3) ASA 500 mg q.i.d. alone; (4) placebo q.i.d. alone. Two-way analysis of variance showed functional synergism of ASA and scaling, resulting in a therapeutic efficacy approximately equivalent to the sum of each individual therapeutic efficacy (i.e., ASA alone and scaling alone) in reducing gingival inflammation and pocket probing depth over the 6-week observation period (interaction: p > 0.05). Only the effect of ASA was significant in reducing the concentration of elastase-alpha 1-proteinase inhibitor in gingival crevicular fluid (GCF E-alpha 1-PI) (p < 0.001), reduction in GCF E-alpha 1-PI concentrations by ASA may indicate a decreased risk in periodontal disease progression. The results suggest that the combination of therapies and their different mechanisms of action, i.e., reduction of bacterial plaque and inhibition of destructive components of the immune responses, may result in functionally synergistic therapeutic efficacies in patients with untreated adult periodontitis. PMID- 8707973 TI - Effect of localized experimental gingivitis on early supragingival plaque accumulation. AB - Previous investigations have reported increased plaque formation in the presence of gingival inflammation as compared with gingival health. In these investigations, experimental gingivitis has been induced by total abolition of oral hygiene such that increased levels of salivary bacteria could contribute to the increased plaque formation. In the present study, experimental gingivitis was localized to 2 selected teeth whilst maintaining normal oral hygiene measures in the rest of the mouth. The aim of this study was to investigate whether the mass of plaque accumulating in a 3-day period is influenced by the inflammatory status of the adjacent gingival margins. Following scaling, prophylaxis and a period of optimal hygiene to establish gingival health, "plaque-guards" were worn during routine oral hygiene performance to prevent any cleaning on the buccal surfaces of teeth 14 and 15 from days 0 to 3. On day 3, accumulated plaque was removed from a 1.5 mm zone on the buccal surfaces of 14 and 15 adjacent to the gingival margins and weighed immediately. Subjects continued to wear their plaque-guards during oral hygiene until day 14 to induce experimental gingivitis around the experimental teeth. At day 14, all plaque was cleaned from the buccal surfaces of the experimental teeth and the mass of plaque accumulating over the next 3 days weighed on day 17. Comparison of plaque weights showed that the wet-weight of 3 day-old plaque was higher in the presence of experimental gingivitis than in the presence of gingival health (p = 0.02). This observation suggests that the inflammatory status of the marginal gingiva has an important effect on early, supragingival plaque accumulation. PMID- 8707974 TI - The pathogenesis of drug-induced gingival overgrowth. AB - Gingival overgrowth is a well-documented unwanted effect, associated with phenytoin, cyclosporin, and the calcium channel blockers. The pathogenesis of drug-induced gingival overgrowth is uncertain, and there appears to be no unifying hypothesis that links together the 3 commonly implicated drugs. In this review, we consider a multifactorial model which expands on the interaction between drug and/or metabolite, with the gingival fibroblasts. Factors which impact upon this model include age, genetic predisposition, pharmacokinetic variables, plaque-induced inflammatory and immunological changes and activation of growth factors. Of these, genetic factors which give rise to fibroblast heterogeneity, gingival inflammation, and pharmacokinetic variables appear to be significant in the expression of gingival overgrowth. A more thorough understanding of the pathogenesis of this unwanted effect will hopefully elucidate appropriate mechanisms for its control. PMID- 8707975 TI - Systemic retinoid medication and periodontal health in patients with Papillon Lefevre syndrome. AB - Periodontal health in relation to systemic retinoid medication was evaluated retrospectively in patients with Papillon-Lefevre Syndrome (PLS). The material consisted of 18 children/young adults ranging from 8 to 28 years of age, all with a confirmed diagnosis of PLS. 9 participants, comprising a medication group, had been on long-term (range 1.5-9 years) retinoid medication for their cutaneous lesions. The remaining 9 served as controls. Regardless of whether or not retinoid medication was received, every patient experienced an early and devastating periodontitis, with atypical edematous and erythematous gingiva, suppuration from deep gingival pockets and premature loss of teeth. No correlation could be found between the severity of skin involvement and the severity of periodontal involvement. An improvement with age could be seen for the cutaneous lesions but not for the periodontal condition. Systemic medication with retinoids had a favorable therapeutic effect on cutaneous lesions, and no severe complication/side effect could be seen after several years of continuous use. However, from the results of this study it can be concluded that, at least in a situation with poor compliance of daily oral home-care, no positive effect on the periodontal health in patients with PLS could be seen by the retinoid medication. PMID- 8707976 TI - The effect of a single mechanical treatment on the subgingival microflora in patients with HIV-associated gingivitis. AB - The purpose of the present study was to investigate the effect of a single episode of scaling and root planing on clinical periodontal parameters and on the subgingival microflora in human immunodeficiency virus (HIV)-positive and HIV negative subjects. 13 subjects participated and were clinically scored at days 7, 7, 30 and 90. 7 subjects were infected with HIV and presented an HIV associated gingivitis. 6 subjects were HIV-negative with at least 12 teeth affected by conventional gingivitis. No significant differences were seen between both groups regarding the presence of P. gingivalis, P. intermedia and A. actinomycetemcomitans. The mean plaque index (PlI) was 1.79 in the HIV-negative and 1.29 in the HIV-positive group. The mean gingival index (GI) was higher in the HIV-positive group (HIV-positive: 1.55; HIV-negative: 1.47). Whereas, the PlI decreased significantly in the HIV-negative group during the course of the study, no change was observed in the HIV-positive group. GI as well as bleeding upon sampling decreased significantly in both patient groups during the same period. The bacterial counts decreased from day -7 to day 7 and generally remained on a lower level until day 90. Small differences were seen in the microbiological flora of the HIV-positive and the HIV-negative subjects following mechanical treatment. PMID- 8707977 TI - Evaluation of the healing response following placement of Coe-pak and an experimental pack after periodontal flap surgery. AB - 2 periodontal dressings, Coe-pak and RD8 (a product under development), were assessed following periodontal surgery, using a double-blind study of randomised split-mouth design, with each patient acting as their own control. Matched sites were selected for periodontal flap surgery under local analgesia. Sites were randomly allocated to each group. The dressings were applied by a single operator and both patients and clinical assessors were blind as to the dressing employed. The dressings were assessed as to their handling properties, in addition to a 7 day post-operative assessment of the surgical sites and a subjective patient report. Maximum pain levels were relatively low (4 in a scale 0-10); subjects reported more pain and analgesics required with RD8 than with Coe-pak. This was reflected in the results of the patient assessment which indicated that 50% preferred Coe-pak, compared to 25% who preferred the RD8 dressing. The results showed RD8 to be an acceptable periodontal dressing when compared to Coe-pak. There were no statistically significant differences between treated sites in both groups with regard to post-operative plaque scores, bleeding scores or tissue healing. PMID- 8707978 TI - Medical status and complications in relation to periodontal disease experience in insulin-dependent diabetics. AB - The aim of this study was to define a population of diabetics exhibiting an increased risk of developing severe periodontitis by comparing the medical status of 2 groups of diabetics, 1 with no/minor periodontal disease and 1 with severe periodontal disease. The case-control study consisted of 2 parts, a baseline study and a follow-up study. 39 case-control pairs were selected. They were adult, long-duration, insulin-dependent diabetics matched according to sex, age and diabetes duration. One individual in each pair (the CASE) exhibited severe periodontal disease while the other (the CONTROL) exhibited gingivitis or only minor alveolar bone loss. The median age of the cases was 58 years (range 36 to 70 years) and of the controls 59 years (range 37 to 69 years). The median disease duration in cases and controls was 24 years and 25 years, respectively. The median follow-up time was 6 years. The medical variables analysed were weight, insulin dose, systolic and diastolic blood pressure, vibratory threshold, triglycerides, total-cholesterol, HDL-cholesterol, creatinine, HbA1, proteinuria, ECG, retinopathy, stroke, transient ischemic attacks (TIA), angina, myocardial infarct, heart failure, hypertension, intermittent claudication, foot ulcer, death, cause of death, and smoking habit. Biochemical analyses and clinical variables used as a routine in the monitoring of diabetics failed to differentiate between diabetics with severe and minor periodontal disease. In the follow-up study, significantly higher prevalences of proteinuria and cardiovascular complications such as stroke, TIA, angina, myocardial infarct and intermittent claudication were found in the case group. An association between renal disease, cardiovascular complications and severe periodontitis seems to exist. This indicates that a closer cooperation between the diabetologist and the dentist is necessary in monitoring the diabetic patient. PMID- 8707979 TI - Longitudinal changes in antibody avidity to Actinobacillus actinomycetemcomitans in periodontitis. AB - Longitudinal investigations concerning immunological responses and periodontal disease activity support a relationship between serum antibody levels and the micro-organisms associated with dental plaque. To define this host response further, we studied the relationship of antibody avidity to Actinobacillus actinomycetemcomitans (Aa) in 11 adult periodontitis and 6 localized juvenile periodontitis (LJP) patients with Aa infections. Patients were monitored every 3 4 months for immunological and clinical variables including probing pocket depths (PD), bleeding on probing (BOP), plaque index (PLI), and the number of disease active sites (DA). Avidity indices were determined using an ELISA and significant changes from each patients' baseline level were determined. The results showed different response patterns between and within the patient groups. A subset of the subjects experienced significant changes in antibody avidity over time. Between group comparisons yielded no significant differences in the number of positive or negative avidity index changes, although there were more frequent changes in the disease active adult periodontitis group. There were also no significant correlations between clinical parameters and antibody avidity, although there were changes in the clinical parameters between baseline and significant avidity change points, and also between baseline and the determination of active disease. Further studies will be necessary to define fully the role of antibody avidity and its relationship to the pathogenesis of periodontal diseases. PMID- 8707980 TI - Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans and Treponema denticola detection in oral plaque samples using the polymerase chain reaction. AB - Detection of putative pathogens is critical for delineating the etiology and progression of periodontitis. In the present study, we have used a polymerase chain reaction (PCR) assay utilizing primers specific for the lkt A gene of Actinobacillus actinomycetemcomitans, the fimbrial gene of Porphyromonas gingivalis, and tdp A gene of Treponema denticola in order to determine the presence of these pathogens in subgingival plaque samples from periodontitis sites. These gene specific primers were also used to assess the detection of different strains of bacteria in the PCR assays. Primers for P. gingivalis detected P. gingivalis strain 33277, but no product was detected when primers were used with extracts from 4 species of Capnocytophaga, 3 species of Prevotella, 2 species of Porphyromonas other than P. gingivalis, Bacteroides levii, Escherichia coli, 3 strains of A. actinomycetemcomitans and 3 strains of T. denticola. PCR analysis using primers for the lkt A gene of A. actinomycetemcomitans also did not result in a product with any of these bacteria with the exception of a positive result with 3 different strains of A. actinomycetemcomitans. Primers selected from the tdp A gene of T. denticola did not identify any of the bacteria strains tested except T. denticola serovars a, b, and c. Thus, these primers were shown to amplify gene segments that are specific to either P. gingivalis (33277), A. actinomycetemcomitans (33384, 43717 and 43718) or T. denticola (35405, 33521 and 35404). The PCR assay may be used to rapidly detect the presence of periodontal pathogens in the future. PMID- 8707982 TI - Supportive periodontal care. Scientific basis for long-term success. Proceedings of the 7th International Congress of the Italian Society of Periodontology. PMID- 8707981 TI - A 6-month home-usage trial of 0.1% and 0.2% delmopinol mouthwashes (I). Effects on plaque, gingivitis, supragingival calculus and tooth staining. AB - Delmopinol is a morpholinoethanol derivative which, in mouthrinses used in the absence of normal oral hygiene, has been shown effective in the inhibition of plaque and gingivitis. The aim of this study was to determine the adjunctive oral hygiene benefits and safety of delmopinol rinses when used alongside normal toothcleaning. This 6-month home use study was a placebo-controlled, double blind, randomised parallel design evaluating 0.1% and 0.2% delmopinol rinses and structured to conform with the ADA Council of Dental Therapeutics guidelines. A total of 450 dentate male and female subjects were recruited who had no relevant medical or pharmacotherapy histories determined from a full medical examination, including haematological and biochemical tests. Subjects had moderate levels of plaque and gingivitis. At baseline, 3 and 6 months subjects were scored for plaque, gingivitis, tooth stain and supragingival calculus, with plaque sampled for microbiological analysis. Additionally, oral mucosal examinations were performed and subjects questioned for adverse symptoms. Baseline special tests were repeated at the end of the study. After baseline examinations, the subjects received a professional prophylaxis, provided with the allocated mouthwash and instructed to use 10-ml volumes for 60 s 2 x daily and where appropriate after toothbrushing and meals. Demographic features of the 3 groups were similar and losses to trial were small. Adverse signs and symptoms included transitory numbness of the tongue, tooth and tongue staining, taste disturbance and rarely mucosal soreness and erosion. All local side-effects were less commonly reported at 6 compared to 3 months and only 6 subjects were withdrawn because of adverse event. No systemic effects attributable to the agent were observed and no significant shifts in haematological or biochemical parameters occurred. All groups showed considerable improvements in oral hygiene and gingival health with some significant differences in favour of 0.2% delmopinol compared to placebo for gingivitis and more particularly plaque. Staining was also significantly increased in the delmopinol groups but not calculus. In the present study, a considerable Hawthorne effect occurred, which must in part explain why only a modestly significant effect was achieved. PMID- 8707983 TI - Risk assessment of recurrence of disease during supportive periodontal care. Epidemiological considerations. AB - Although it is accepted that the primary cause of periodontitis is bacterial infection of long duration, there are a number of risk factors which may increase the probability of recurrence of periodontal disease during supportive periodontal care. The risk may in such cases be caused by other factors than poor oral hygiene measures per se. Cross-sectional and longitudinal studies show conflicting results concerning age as a risk factor for periodontal disease. The effect of smoking on the periodontal tissues has been discussed for decades and only lately has it been possible to demonstrate that smokers definitely have more periodontal problems than non-smokers. Another important risk factor for periodontitis relates to the insulin dependent and non-insulin dependent forms of diabetes mellitus. Poorly-controlled long-duration diabetics have more periodontitis and tooth loss than well-controlled or non-diabetics. Finally, the issue of compliance deserves attention. The medical literature has suggested that patients with chronic illnesses tend to comply poorly, especially if the disease is not perceived to be particularly threatening, if the therapy is time consuming, or if the symptoms are non-disturbing. Suggestions for improved compliance are called for. PMID- 8707984 TI - Periodontal diagnosis in treated periodontitis. Why, when and how to use clinical parameters. AB - The objective of clinical periodontal diagnosis in maintenance patients is to monitor the risk for periodontal disease progression. Risk for progression should be continuously monitored at the patient, tooth and site level at each recall appointment. At the patient level, the significance of systemic diseases, cigarette smoking, compliance with the recall program, loss of support in relation to the patient's age, full mouth plaque and/or bleeding scores, and prevalence of residual pockets are of key importance. At the tooth and tooth-site levels, residual periodontal support, inflammatory parameters and their persistence, presence of ecological niches with difficult access such as furcations, and presence of iatrogenic factors have to be put into proportion with the patient's overall risk profile. The information gathered by clinical monitoring and continuous multilevel risk assessment facilitates an immediate appreciation of the periodontal health status of an individual and the possible risk for further infection and/or disease progression in the dentition and at a particular tooth or site. PMID- 8707985 TI - Microbiological monitoring. AB - If periodontal disease is due to a limited number of bacterial species, then continuous maximal plaque suppression is not the only possibility for prevention and therapy. Specific elimination or reduction of pathogenic bacteria from plaque becomes a valid alternative. Recent studies indicate that the elimination of certain putative pathogens is particularly difficult. New diagnostic methods should allow the choice of better suited procedures, make chosen procedures more effective (through better timing, dosage, selection of devices or drugs, increase of specificity, etc.) or lead to the elimination of unnecessary work (e.g., the treatment of non-susceptible sites or patients). The benefit of newly proposed tests depends on the possible impact of the obtained information on clinical decisions and on the consequences these decisions have for treatment. Thus, diagnostic methods and therapeutical options have to be evaluated together. PMID- 8707986 TI - Host response assessment in recurring periodontitis. AB - Data derived from periodontitis patients have provided support for a relationship between the distribution of selected members of the periodontopathic microbiota and antibody levels to the intact bacteria in both serum and GCF. These data are consistent with the systemic antibody as a reflection of the host response to an infectious process associated with an episode of disease activity. The purpose of this report is to address the concept that the host antibody responses may help to elucidate the specific etiologic agents and be used to model the risk for future periodontal disease progression in recurring periodontitis. These findings from one study in adult periodontitis patients indicated that elevations in certain antibody specificities are most closely associated with patients exhibiting a risk of disease recurrence. Furthermore, analysis of the frequency of antibody elevations suggested that patients capable of maintaining elevated antibody to these pathogens post-treatment, may be indicative of an individual at less risk. A 2nd investigation was implemented to address questions concerning host-parasite interactions in A. actinomycetemcomitans-associated recurring periodontitis. The results showed distinctive characteristics of local and systemic antibody responses and A. actinomycetemcomitans infection in patients with varying extents of recurrent disease. These longitudinal studies developed evidence for the potential of local and/or systemic antibody responses as indicators of periodontal disease recurrence. PMID- 8707987 TI - Rationale of mechanical plaque control. AB - Patients who have received extensive periodontal treatment also demonstrate a high susceptibility to periodontal disease. Maintenance of periodontal health following therapy includes a lifelong supportive care consisting of daily removal of the microbial plaque by the patient, supplemented by professional care in an individually designed programme. Mechanical supragingival plaque control by self care is of utmost importance. The goal is to create a positive attitude by information and motivation to give the patient knowledge and confidence. The patient should be advised to use appropriate aids and technique. A soft brush, an interspace brush, interdental tooth brushes or tooth picks are recommended in periodontal patients. Professional tooth cleaning involves removal of supragingival plaque from all tooth surfaces using mechanically driven instruments and fluoride prophy paste and, when indicated, removal of calculus and subgingival plaque. Disclosing solution is used to visualize the plaque to the patient and to the clinician in order to reinforce instruction in oral hygiene. Oral hygiene measures alone seem to have limited effect on subgingival microflora in cases of severe disease. In shallow and moderately deep pockets a good plaque control can change the subgingival flora towards a more "healthy" composition. Subgingival plaque removal is performed with hand- and/or ultrasonic instruments. Cracks within the cementum, grooves, fissures, resorption lacunae, furcations may create difficulties in cleaning the root surface. Ultrasonic instrumentation has a beneficial effect in creating a smooth surface without extensive removal of cementum. Besides, the cavitational activity contributes to plaque removal which makes the instrument further suitable during maintenance therapy. The result of the debridement is assessed on the healing response in the tissues. The frequency of maintenance visits must be given on an individual basis according to the needs of every special patient. The visit includes plaque evaluation (disclosion), oral hygiene instruction, probing depth measurements, registration of bleeding on probing, scaling (plaque removal) if indicated, tooth polishing, fluoride application and radiographs if indicated. The goal is to identify and treat signs of recurrence of periodontal disease in order to prevent further loss of attachment. PMID- 8707988 TI - Principles of pharmacologic intervention. AB - Compared to studies with conventional maintenance, relatively few studies have been conducted that have tested pharmacologic intervention as a means for supportive periodontal care. Despite the scarcity of data, several principles have begun to emerge. First, it is clear that removal of subgingival calculus is necessary for the highest level of a long-term effectiveness. Hence, pharmacologic intervention appears best suited as an adjunctive therapy directed toward "problem sites"; sites that fail to respond adequately to conventional maintenance procedures. Intrapocket drug delivery systems appear to offer particular promise since therapy can be directed to selected sites that appear to be failing. Additional effectiveness may also be obtained by use of chlorhexidine mouth rinses for short periods of time during healing to control re-infection. Eradication of reservoirs of infection throughout the mouth also appears to be an important principle related to long-term stabilization. Studies to date suggest that superior clinical response can be obtained by intrapocket delivery systems. Furthermore, up to 2 years of periodontal stabilization can be achieved by these means and longer disease-free maintenance intervals can be established. Additional clinical trials will be necessary to fully optimize and understand this therapeutic approach, but initial studies have reported promising results. PMID- 8707989 TI - Long-term periodontal maintenance in private practice. AB - Adult periodontitis is a chronic disease suffered by the mainstream of the world's population. To date, its treatment consists of providing support, control, and stabilization of the disease rather than a cure. Although treatment modalities rely upon a maintenance program to preserve a controlled environment that has been established, it is painstakingly evident that total patient compliance does not exceed 40% in private practice. In addition, there is little evidence that any society is populated by plaque-free patients. The important variables in private practice are the end point goals of treatment, manpower availability, patient behavior, the size of the treatment facility and the economic pressures of the geographic location. Each of these has to be considered in its entirety to appreciate the difficulties encountered in managing this significant issue in a periodontal practice. PMID- 8707990 TI - Cost effectiveness of periodontal services. AB - This paper presents the combined use of decision trees and cost-effectiveness analysis to determine on which patients new diagnostic tests can be useful, and whether they can change the dentist's decision to provide different services and thus improve patient outcomes in 5 different categories of patients. A decision tree is constructed in which the primary decision branches are "test" and "no test". The treatment and outcome branches in the test branch are affected by the results of the test which is then either negative or positive. In contrast, the treatment and outcome branches in the no-test branch represent pathways for patient therapy when the new (or additional) test is not used. Results show that even with excellent test efficacy parameters (sensitivity 0.87 and specificity 0.84) it is not cost effective to use a diagnostic test in the general population or for adults under age 45 years. However for adults > 45 years, patients referred to a periodontist, and patients with a history of periodontal disease, the test would change treatment decisions and improve patient care outcomes. PMID- 8707991 TI - Psychological intervention in patients with poor compliance. AB - In this article, the authors examine the compliance of periodontal patients from a behavioral psychology perspective. The causes of non-compliance are analyzed along with the main methods of intervention. An experiment was carried out in which various behaviour modification techniques were compared. This study, in particular, examined the effect of periodontist's use of behaviour modification techniques on patient compliance in four treatment maintenance conditions. A functional analysis of compliance and non-compliance behaviour revealed various treatment opportunities. Hence, 4 treatment conditions (which included basic oral hygiene instructions, performance feedback, behavioural self-management, and positive reinforcement) were compared in 3 phases. Data collected over a period of 3 months indicated that behavioural self-management plus positive reinforcement increase patient compliance significantly. Implications for further research are discussed in terms of analyzing the cost of each component procedure for both the patient and the doctor in terms of time, money and effort required. Hence, the present study demonstrates that behavioural strategies can make a significant contribution to the area of patient management. PMID- 8707992 TI - Refractory periodontitis or recurrence of disease. AB - Refractory periodontal disease is characterized by low plaque scores and low responsiveness to periodontal therapy. The patients often have a history of antibiotic therapy and therefore have a high incidence of resistance in the subgingival microflora. The above features are in contrast to adult chronic periodontal disease and recurrence of disease. The subgingival microflora of refractory disease may be either predominantly gram-positive with elevated levels of Streptococcus intermedius or may be gram negative and contain elevated levels of the classical periodontal pathogens. In some cases serum IgG antibodies against Porphyromonas gingivalis are elevated and seem to decrease after therapy. Smoking may be an important factor in refractory periodontal disease. Treatment with directed antibiotic therapy as an adjunct to scaling and root planing retards the progression of disease for a limited time. Many questions are still not answered and suggestions are made for future directions in research. PMID- 8707993 TI - Refractory periodontitis: critical questions in clinical management. AB - Refractory periodontitis appears to be a patient phenomenon with a clinical pattern that may be the result of different influences including genetic factors and exogenous factors that alter the host response to the bacterial challenge. Different patients may have different mechanisms involved in their disease and therefore may show different response patterns following therapy. Although localized non-responsive sites in otherwise responsive patients should not be considered refractory periodontitis, such sites do present substantial therapeutic problems. Initial evidence suggests that local delivery tetracycline provides significant advantages in the management of such sites. Although refractory periodontitis cases appear to be primarily the result of host alterations, the most predictable approach to therapy at present involves systemic antibiotics in conjunction with local therapy. The goal of antibiotic therapy should be to eliminate from detection the target bacteria that were present prior to therapy. Good plaque control, regular professional maintenance therapy, regular microbial monitoring, and control of risk factors are important aspects of the long term maintenance of these cases. PMID- 8707994 TI - The impact of new technologies to diagnose and treat periodontal disease. A look to the future. AB - The last 25 years have brought unprecedented advances to our understanding of periodontal disease. Consider that in 1970 periodontitis was believed to effect most individuals over the age of 35 years, to progress steadily in an individual once initiated until teeth were lost, to be the primary cause of tooth loss in adults, to be caused by the bacterial mass accumulating on the tooth surface and subgingivally, and to involve the host in some fashion or another. In the 25 years since then, impressive research advances in the epidemiology of periodontal disease, the specific bacterial etiology of periodontal disease and the immunoinflammatory mediators of periodontal tissue destruction have greatly altered our view of periodontal disease. Thus, given these research advances in the understanding of periodontitis, what may the future hold for improved diagnosis and treatment of periodontal disease? Impressive research into new ways to diagnose the periodontal diseases is well underway. Investigators are seeking new ways to diagnose an individual's degree of risk for periodontal disease initiation, susceptibility to disease progression, level of disease "activity" and the likely response to treatment and recurrence of active disease. New diagnostic tests should greatly advance our ability to more accurately and specifically diagnose periodontal disease. The future also looks promising for new treatment strategies to slow or arrest periodontal disease progression. The bacterial specificity of periodontal disease etiology revealed since 1970 has logically led to the use of antibiotics in periodontitis treatment. In the late 1980s the concept of locally delivering antibiotics to the periodontal pocket was introduced, and subsequent clinical trials have indicated that it is possible to reduce pocket depth and inflammation with tetracycline locally delivered to the periodontal pocket. Likely, we have barely scratched the surface in studying the efficacy of locally delivery antimicrobial agents to alter the progression of periodontal disease. As new agents are developed and better delivery systems to the periodontal pocket are developed, the future should see a variety of antimicrobial agents available which can slow periodontal disease progression. The future also holds promise for slowing periodontal disease progression by blocking inflammatory pathways important in periodontal tissue destruction. Clinical trials of flubiprofen, naproxen and ketoprofen indicate that it is possible to slow periodontal disease progression with non-steroidal anti inflammatory drugs which inhibit one destructive pathway. In addition, data from animal models indicate that chemically modified tetracycline as an inhibitor of collagenase can slow disease progression in animals. Again, we have likely only just begun to explore the wide range of molecular mediators of tissue destruction which may be targeted for blocking and thereby slow or arrest periodontal disease progression. Last, research into regenerating periodontal structures lost as a result of disease has had a noteworthy record of progress in the past 25 years. Techniques that utilize bone grafts, root treatments, tissue guiding membranes or polypeptide growth factors have ably indicated that it is possible to regenerate new attachment structures in humans. As investigators continue to unravel the mysteries of the embryonic development of the periodontium, the ability to predictably regenerate lost periodontal attachment structures holds great promise for the future. PMID- 8707995 TI - Key issues in the diagnosis and treatment of bipolar disorders. PMID- 8707996 TI - Outcome in the pharmacologic treatment of bipolar disorder. AB - Outcome studies of bipolar disorder, the majority of which were conducted before the use of lithium, divalproex, and carbamazepine, generally found that only 50 to 60% of patients achieved good recovery 6 to 12 months after a manic episode. Over the past decade, a number of new pharmacologic studies have provided further information regarding the acute and long-term outcome of patients with bipolar disorder treated with these medications. In addition, better operational criteria to define outcome have been advanced, allowing for easier extrapolation of the results of clinical trials to clinical practice. We review the outcome of studies of lithium, divalproex, and carbamazepine in the acute treatment of episodes of mania and bipolar depression and in the maintenance treatment of bipolar disorder and the implications of these studies to clinical practice. PMID- 8707997 TI - Predictors of response to mood stabilizers. AB - Although lithium remains the preferred treatment of bipolar disorder, only 60 to 80% of patients with the classic presentation have an adequate response to this drug. When the response rate to lithium is considered across the entire spectrum of bipolar disorders, this rate probably decreases to 50%. Natural history, illness subtype, and comorbidity are all important general predictors of response to treatment. At present, the only predictors that seem to differentially favor divalproex, and possible, carbamazepine over lithium are mixed states and rapid cycling. An overview of clinical presentations that predict general and differential response to mood stabilizers is provided. PMID- 8707998 TI - Bipolar mood disorder: practical strategies for acute and maintenance phase treatment. AB - The chronic, complex, and episodic course of bipolar mood disorder presents a formidable challenge to the clinician making a treatment plan. Although numerous therapeutic agents are reported to be efficacious for one or more aspects of bipolar illness, treatment is seldom completely effective and is never curative. The goal of treatment is to modify the symptomatic expression of the illness with the result that fewer, briefer, and milder episodes occur. In pursuit of this goal, the use of multiple medications, polypharmacy, is the rule rather than the exception. This article offers recommendations for treatment strategies based on the phase and stage of the illness. The strategies are organized into algorithms that emphasize the use of mood-stabilizing medications as initial steps in a systematic, iterative approach to treatment. Practical issues related to the use of mood-stabilizing therapies are discussed. PMID- 8707999 TI - The prevalent clinical spectrum of bipolar disorders: beyond DSM-IV. AB - Based on the author's work and that of collaborators, as well as other contemporaneous research, this article reaffirms the existence of a broad bipolar spectrum between the extremes of psychotic manic-depressive illness and strictly defined unipolar depression. The alternation of mania and melancholia beginning in the juvenile years is one of the most classic descriptions in clinical medicine that has come to us from Greco-Roman times. French alienists in the middle of the nineteenth century and Kraepelin at the turn of that century formalized it into manic-depressive psychosis. In the pre-DSM-III era during the 1960s and 1970s, North American psychiatrists rarely diagnosed the psychotic forms of the disease; now, there is greater recognition that most excited psychoses with a biphasic course, including many with schizo-affective features, belong to the bipolar spectrum. Current data also support Kraepelin's delineation of mixed states, which frequently take on psychotic proportions. However, full syndromal intertwining of depressive and manic states into dysphoric or mixed mania--as emphasized in DSM-IV--is relatively uncommon; depressive symptoms in the midst of mania are more representative of mixed states. DSM-IV also does not formally recognize hypomanic symptomatology that intrudes into major depressive episodes and gives rise to agitated depressive and/or anxious, dysphoric, restless depressions with flight of ideas. Many of these mixed depressive states arise within the setting of an attenuated bipolar spectrum characterized by major depressive episodes and soft signs of bipolarity. DSM-IV conventions are most explicit for the bipolar II subtype with major depressive and clear-cut spontaneous hypomanic episodes; temperamental cyclothymia and hyperthymia receive insufficient recognition as potential factors that could lead to switching from depression to bipolar I disorder and, in vulnerable subjects, to predominantly depressive cycling. In the main, rapid-cycling and mixed states are distinct. Nonetheless, there exist ultrarapid-cycling forms where morose, labile moods with irritable, mixed features constitute patients' habitual self and, for that reason, are often mistaken for "borderline" personality disorder. Clearly, more formal research needs to be conducted in this temperamental interface between more classic bipolar and unipolar disorders. The clinical stakes, however, are such that a narrow concept of bipolar disorder would deprive many patients with lifelong temperamental dysregulation and depressive episodes of the benefits of mood-regulating agents. PMID- 8708000 TI - Role of newer medications for bipolar disorder. AB - The quest for additional effective therapies for bipolar study commenced in the 1960s, even before regulatory approval of lithium. This was driven by the recognition that patient response to lithium varied from excellent to poor; thus, significant numbers of patients faced its frequent adverse effects and risks without sustaining any benefit. These needed advances have been slowed by the unusual difficulties of studying bipolar disorder in controlled, rigorously designed protocols. These difficulties are reviewed both from the perspective of their effects on research and on clinical practice. In addition, the results of two blinded, placebo-controlled studies of divalproex in the treatment of acute mania are compared with results from earlier open and comparator studies. The comparisons indicate that open-study results with divalproex and other forms of valproic acid have yielded results quite similar to those observed under rigorous study conditions. Whereas substantial evidence exists regarding the efficacy of divalproex and carbamazepine and their roles in treating patients with bipolar disorder, studies evaluating other putative mood-stabilizing agents and antidepressants in bipolar disorder are few in number and inconsistent in results. With few exceptions, studies of these latter drugs do not provide clear guidelines for treatment. PMID- 8708001 TI - Psychotherapy in combination with drug treatment for bipolar disorder. AB - There is a new movement toward delivering manualized, empirically validated psychotherapies in combination with pharmacologic maintenance in the outpatient treatment of bipolar disorder. This article describes the specific goals of psychotherapy with bipolar patients: modifying social risk factors, enhancing medication adherence, increasing the patient's and family's willingness to accept the reality of the disorder, and reducing suicidal risk. It also reviews the nature and underlying assumptions for two new psychotherapies: a family psychoeducational treatment and an individual treatment. Problems in delivering new treatments in the era of managed care, and some solutions to these problems, are also discussed. PMID- 8708002 TI - D1 dopamine receptor-mediated induction of zif268 and c-fos in the dopamine depleted striatum: differential regulation and independence from NMDA receptors. AB - Excitatory amino acid afferents from cerebral cortex and dopamine afferents from the substantia nigra synapse on common projection neurons in the striatum. Activation of D1 dopamine receptors increases immediate early gene expression in the striatum and conductance through the N-methyl-d-aspartate (NMDA) receptor. To examine the contribution of NMDA receptor activation to dopamine receptor mediated responses, we determined the effects of intrastriatal administration of NMDA antagonists on immediate early gene expression in the striatum and rotational behavior induced by stimulation of the D1 receptor in rats with unilateral dopamine depletions. Systemic administration of SKF 38393 increased c fos and zif268 mRNAs in the striatum and induced contralateral rotation. Intrastriatal infusion of the competitive NMDA receptor antagonist (+/-)-3-(2 carboxypiperazin-4-yl)-propyl -1-phosphonic acid caused a dose-dependent attenuation of SKF 38393-induced rotation and partially decreased c-fos mRNA expression. However, D1-mediated increases in zif268 mRNA were not affected, except by the highest concentration of antagonist used (10 mM). Another competitive antagonist, 2-amino-5-phosphonovaleric acid, had similar effects. Like the competitive antagonists, intrastriatal infusion of the non-competitive NMDA antagonist MK-801 partially decreased c-fos, but not zif268, mRNA in the area around the microdialysis probe. However, unlike competitive antagonists, local infusion of 1 mM MK-801 potentiated D1-mediated increases in c-fos and zif268 mRNAs in lateral striatum. These data suggest that 1) some D1 dopamine receptor-mediated effects on striatal function are independent of ongoing NMDA receptor activation, whereas other effects are at least partially mediated by NMDA receptor activity in the striatum, and 2) competitive and non-competitive antagonists of the NMDA receptor differently affect D1-mediated immediate early gene expression in the striatum. PMID- 8708003 TI - Developmental and plasticity-related differential expression of two SNAP-25 isoforms in the rat brain. AB - In this article we study the relationship between the expression pattern of two recently identified isoforms of the 25-kD synaptosomal-associated protein (SNAP 25a and SNAP-25b) and the morphological changes inherent to neuronal plasticity during development and kainic acid treatment. SNAP-25 has been involved in vescicle fusion in the nerve terminal, and most likely participates in different membrane fusion-related processes, such as those involved in neurotransmitter release and axonal growth. In the adult brain, SNAP-25b expression exceeded SNAP 25a in distribution and intensity, being present in most brain structures . Moderate or high levels of SNAP-25a hybridization signal were found in neurons of the olfactory bulb, the layer Va of the frontal and parietal cortices, the piriform cortex, the subiculum and the hippocampal CA4 field, the substantia nigra/pars compacta, and the pineal gland, partially overlapping SNAP-25b mRNA distribution. In restricted regions of cerebral cortex, thalamus, mammillary bodies, substantia nigra, and pineal glands the two isoforms were distributed in reciprocal fashion. During development SNAP-25a mRNA was the predominant isoform, whereas SNAP-25b expression increased postnatally. The early expression of SNAP 25a in the embryo and the decrease after P21 is suggestive of a potential involvement of this isoform in axonal growth and/or synaptogenesis. This conclusion is indirectly supported by the observation that SNAP-25a mRNA, but not SNAP-25b mRNA, was upregulated in the granule cells of the adult dentate gyrus 48 hours after kainate-induced neurotoxic damage of the hippocampal CA3-CA4 regions. Increase of SNAP-25 immunoreactivity was observed as early as 4 days after kainate injection within the mossy fiber terminals of the CA3 region, and in the newly formed mossy fiber aberrant terminals of the supragranular layer. These data suggest an isoform-specific role of SNAP-25 in neural plasticity. PMID- 8708004 TI - Innervation of VIP-immunoreactive neurons by the ventroposteromedial thalamic nucleus in the barrel cortex of the rat. AB - We investigated the synaptic terminals of fibers originating in the ventroposteromedial thalamic nucleus (VPM) and projecting to the main input layers (IV/III) of the rat posteromedial barrel subfield. It was our aim to determine whether or not the subpopulation of vasoactive intestinal polypeptide (VIP)-immunoreactive neurons in these layers are directly innervated by the sensory thalamus. Anterograde tracing with Phaseolus vulgaris leucoagglutinin (PHA-L) and immunohistochemistry for VIP were combined for correlated light and electron microscopic examination. Columns of cortical tissue were well defined by barrel-like patches of PHA-L-labeled fibers and boutons in layers IV and III. Within these columns VIP-immunoreactive perikarya were located mainly in supragranular layers. Marked perikarya were also seen in infragranular layers, but their immunoreactivity was often weaker. Granular layer IV, which is the main terminal field for thalamic fibers, contained fewer VIP neurons than supragranular layers. In the light microscope, however, PHA-L-labeled fibers appeared to contact the somata or proximal dendrites of 60-86% of the layer IV VIP neurons . By contrast, only 18-35% of the VIP neurons in the supragranular layers, which receive a moderately dense projection from the VPM, appeared to be contacted. PHA-L-labeled boutons were seen close to 13-25% of infragranular VIP positive cells. Electron microscopy showed that thalamic fibers formed at most four asymmetric synapses on a single layer IV, VIP-positive neuron. Although the proportion of VIP-positive neurons with labeled synapses was lower in supragranular layers, most of them shared multiple asymmetric synapses with labeled thalamic fibers. Up to six labeled synapses were seen on individual VIP neurons in layer III. We conclude that subpopulations of VIP-immunoreactive neurons, located in layers IV, III, and II are directly innervated by the VPM. These neurons may be involved in the initial stages of cortical processing of sensory information from the large, mystacial vibrissae. Since VIP is known to be colocalized with the inhibitory transmitter GABA, it is likely that VIP neurons participate in the shaping of the receptive fields in the barrel cortex. PMID- 8708005 TI - Structure and function of gustatory neurons in the nucleus of the solitary tract: II. Relationships between neuronal morphology and physiology. AB - This study employed intracellular recording and labeling techniques to examine potential relationships between the physiology and morphology of brainstem gustatory neurons. When we considered the neuronal response to the four "prototypic" tastants, we were able to demonstrate a positive correlation between breadth of responsiveness and the number of dendritic branch points. An analysis of the response to eight tastants also revealed an association between dendritic spine density and the breadth of responsiveness, with more narrowly tuned neurons exhibiting more spines. Interestingly, a neuron's "best response" was a relatively poor predictor of neuronal morphology. When we focused on those neurons that responded to only one tastant, however, a number of potentially important relationships became apparent. We found that the cells that only responded to quinine were smaller than the neurons that only responded to NaCl, HCl, or sucrose. The HCl-only neurons, however, were more widespread in the rostrocaudal dimension that the neurons that only responded to NaCl. A number of additional structure-function relationships were identified when we examined the neuronal response to selected tastants. We found that neurons that responded to sucrose but not quinine, as well as neurons that responded to quinine but not sucrose, were more widespread in the mediolateral dimension than neurons that responded to both sucrose and quinine. We also discovered that the neurons that responded to NaCl, but not to NH4Cl or KCl, were larger than neurons that responded to all three salts. We believe that these results support the hypothesis that there are relationships between the structure and function of gustatory neurons in the nucleus of the solitary tract, with the data highlighting the importance of three themes: 1) the relationship between dendritic specializations and tuning, 2) the relationship between dendritic arbor orientation and response properties, and 3) the potential importance of stimulus specific neurons. PMID- 8708006 TI - Common and distinct fusion proteins in axonal growth and transmitter release. AB - We have used the proteolytic properties of botulinum and tetanus neurotoxins (BoNT, TeNT) to cleave three proteins of the membrane fusion machinery, SNAP-25, VAMP/synaptobrevin, and syntaxin, in developing and differentiated rat central neurons in vitro. Then, we have studied the capacity of neurons to extend neurites, make synapses, and release neurotransmitters. All the toxins showed the expected specificity with the exception that BoNT/C cleaved SNAP-25 in addition to syntaxin and induced rapid neuronal death. In developing neurons, cleavage of SNAP-25 with BoNT/A inhibited axonal growth and prevented synapse formation. In contrast, cleavage of VAMP with TeNT or BoNT/B had no effects on neurite extension and synaptogenesis. All the toxins tested inhibited transmitter release in differentiated neurons, and cleavage of VAMP resulted in the strongest inhibition. These data indicate that SNAP-25 is involved in vesicle fusion for membrane expansion and transmitter release, whereas VAMP is selectively involved in transmitter release. In addition, our results support the hypothesis that synaptic activity is not essential for synapse formation in vitro. PMID- 8708007 TI - Morphology of neuroglia in the antennal lobes and mushroom bodies of the brain of the honeybee. AB - We investigated the distribution and anatomical organization of glial cells in the antennal lobes and mushroom bodies of the honeybee. Reconstructions from serial sections, prepared according to the ethyl gallate method, revealed the entire morphology of glial cells in neuropiles, tracts, and the soma rind. The distribution of the glial cell bodies in the neuropiles was derived from the staining of cell nuclei with a fluorescent dye. There are glial cells of different shape in the soma rind which are wrapped around the neuronal cell bodies of the antennal lobes and the Kenyon cells of the mushroom bodies. Glial cells surround neuropilar areas such as the external and lateral sides of the glomeruli of the antennal lobes. Whereas we could not detect glia in the glomerular neuropile, glial cells with long processes are located in the core of the antennal lobe. Extensions of these glial cells also invade tracts containing the olfactory projection neurons. A layer of glial cells separates the mushroom body neuropile from the surrounding protocerebral neuropile. The neuropile of the mushroom bodies is clearly compartmented by glial cells. There is a high density of astrocyte-like glia in a column of the pedunculus which can be followed to the ventral part of the alpha-lobe. A network of mushroom body intrinsic glial cells separates the alpha-lobe from the beta-lobe and the pedunculus. This anatomical description of glial cell types in olfactory information processing pathways of an insect brain provides a framework for further physiological studies of neuroglia in dissociated cell culture. PMID- 8708008 TI - Genesis and fate of the perireticular thalamic nucleus during early development. AB - A striking feature of the internal capsule during early development is that it is full of small neurones. Later, this group of neurones, called the perireticular thalamic nucleus, appears to have reduced in size, and only a few scattered cells are seen. In an effort to understand better the developmental history of the perireticular nucleus this study examines: i) the period of cell generation in the nucleus, ii) the magnitude of cell loss in the nucleus, and iii) the subsequent fate of cells in the nucleus during development. The perireticular cells are generated very early in development, being among the first generated in the thalamus (rats: E13-14; cats: E21-30). In rats, the first perireticular cells are generated at about the same developmental stage as the first subplate cells, which are among the first generated cells of the cortex: in cats, the first perireticular cells are generated well before those in the subplate (E24-30). In rats, the number of perireticular cells during developmental peaks at P5 (approximately 30,000) and then declines sharply (approximately 98%) by P15 (approximately 750), when adult-like patterns are seen. This dramatic loss of perireticular cells is due to both cell death and a migration of cells into the adjacent globus pallidus. The majority of the perireticular cells which migrate into the globus pallidus, however, are likely to die also. The presence of pyknotic profiles (indicators of dying cells) in the rat perireticular nucleus points to cell death as a contributor to the reduction in cell number during development. In this study, a period of relatively high pyknotic profile incidence (number of pyknotic cells per 1,000 "living" cells) is recorded in the perireticular nucleus over a 5 day period, from P2 to P7 (13.5-15.5). Similar values and patterns are recorded in the reticular nucleus and globus pallidus, except that in these structures, a period of relatively high pyknotic profile incidence (15-20) occurs over a shorter period (3 days; P2-5). Previous studies have suggested that some perireticular cells migrate into and settle within the adjacent globus pallidus. This study, with the use of long-term survivals after tracer injections in rats, shows that none (or very few) of these perireticular cells which migrate into the globus pallidus survive into more mature postnatal stages. Tracer (biotinylated dextran) was injected into the sensory nuclei of the dorsal thalamus at early stages (P7) and the rats were allowed to survive for either a day thereafter (to P8) or until well after the period of cell death was complete (to P16 or P21). In the short-term survivals (to P8), there are many dextran-labelled cells seen in the globus pallidus and in the perireticular nucleus. In the long-term survivals (to P16 or P21), by contrast, there are no dextran-labelled cells apparent in the globus pallidus or in the perireticular nucleus. It is likely that these cells in the globus pallidus, as with those in the perireticular nucleus, undergo cell death during development. PMID- 8708009 TI - Localization of enkephalin immunoreactivity in the spinal cord of the long-tailed ray Himantura fai. AB - Enkephalin-like immunoreactivity (ENK-LI) was found throughout the spinal cord of the long-tailed ray Himantura fai. The densest ENK-LI was in the superficial portion of lamina A of the dorsal horn. Lamina B and the deeper parts of lamina A contained radially oriented, labelled fibres. Laminae C, D, and E contained many longitudinally oriented fascicles which were surrounded by a reticulum of transversely oriented, labelled fibres, some of which projected into the ventral and lateral funiculi. Labelled fibres were found in the dorsal commissure and around the central canal, but the later did not cross the midline. One-third of all enkephalinergic cells were found throughout laminae A and B, while two-thirds were located in the medial half of C, D, and E. Occasionally a labelled cell was located in the lateral funiculus. The ventral horn (laminae F and G) contained many enkephalinergic fibres but no labelled nuclei. A few dorsal column axons contained ENK-LI. In the lateral funiculus there were two groups of labelled axons, a superficial, dorsolateral group, and a deeper group, occupying a crescent-shaped region. The ventral funiculus also contained many labelled axons. The central projection of the dorsal root passed through the substantia gelatinosa and divided into rostrally and caudally projecting fascicles within lamina C. The root, and these fascicles, both lacked ENK-LI. In contrast, the fascicles in laminae D and E did contain enkephalinergic fibres. The origin of the various fibre systems and the role of enkephalin in the regulation of sensory processing and motor output are discussed. PMID- 8708010 TI - Human fetal hippocampal development: I. Cytoarchitecture, myeloarchitecture, and neuronal morphologic features. AB - To characterize better the process of anatomic development of the human hippocampus, we studied the cytoarchitecture, myeloarchitecture, and neuronal morphology in human fetal and postnatal hippocampi. Twenty cases were studied in which the ages ranged from 9 weeks gestation through 62 years. Fixed, paraffin embedded, hippocampal sections were stained with cresyl violet for Nissl substance and immunolabeled for myelin basic protein. The hippocampal region at 9 weeks contains 4 layers: a ventricular zone, an intermediate zone, a homogeneous appearing hippocampal plate comprised of bipolar-shaped neurons, and a wide marginal zone. At 15-19 weeks, individual subfields can be distinguished. A distal-to-proximal gradient of cytoarchitectural and neuronal morphologic maturity is seen, with the subiculum appearing more developed than the ammonic subfields and the dentate gyrus appearing least mature. Within each subfield, an "inside-out" gradient of maturity is also evident. By 32-34 weeks gestational age, neurons in CA2 and CA3 have undergone rapid enlargement and morphologic maturation, surpassing CA1, which still contains some immature neurons. The dentate gyrus is the latest area to develop, only assuming a mature cytoarchitecture after 34 weeks. The essential cytoarchitectural appearance of the hippocampal subfields is stable after birth, although there is progressive neuronal enlargement and a decrease in neuronal density throughout childhood into adulthood. Myelination is first evident near term, with strong myelin basic protein immunoreactivity present in the angular bundle, alveus, and fimbria and relatively scant immunoreactivity in the nascent perforant pathway. Myelination in the hippocampus increases in childhood until adolescence, after which the pattern remains unchanged. These studies delineate normal neuroanatomic development and can be used to understand better the mechanisms underlying human neurodevelopmental and neurodegenerative disorders of the hippocampal formation. PMID- 8708011 TI - Human fetal hippocampal development: II. The neuronal cytoskeleton. AB - In this study of the developing human hippocampus, we monitor the timing of onset and the sequential patterns of expression of 11 developmentally regulated proteins that are important components of the neuronal cytoskeleton. Immunohistochemistry using well-characterized antibodies was conducted with fixed paraffin-embedded sections from hippocampi at various stages of fetal and postnatal development. At 9 weeks gestational age, immunoreactivity was evident for the microtubule-associated proteins (MAPs), MAP2 and MAP5, low molecular weight (Mr) neurofilament (NF) protein (NF-L), poorly phosphorylated mid-Mr NF protein (NF-M/P-), vimentin, and alpha-and beta-tubulins within the somatodendritic domain of neurons of the hippocampal plate. Weak immunoreactivity for moderately phosphorylated, high Mr NF protein (NF-H/P + + +), tau, and nestin was observed. Highly phosphorylated mid-Mr NF protein (NF-M/P + + +) and alpha internexin were first detected at 15 weeks and highly phosphorylated, high Mr NF protein (NF-H/P+3) at 20 weeks. At 15 weeks, MAP2, MAP5, and tubulins were expressed in an "inside-out" gradient and in a gradient between hippocampal subfields with subiculum > ammonic subfields > dentate gyrus. These gradients paralleled the maturational gradients seen in cytoarchitectural and neuronal morphologic studies. The adult pattern of neuronal cytoskeletal protein expression in the hippocampus was attained by the second postnatal year for all proteins. Our findings demonstrate an elaborate orchestration of cytoskeletal protein expression within the hippocampus that is qualitatively similar to what is seen in other brain regions and in nonhuman species but which also has some important differences in timing and pattern. The differences in the developmentally regulated expression of neuronal cytoskeletal proteins in separate regions of the central nervous system (CNS) suggest that there are region-specific differences in composition and function of the neuronal cytoskeleton. These observations have implications for understanding the role of the neuronal cytoskeleton in the developing, mature, and diseased CNS. PMID- 8708012 TI - Identification, distribution, and developmental changes of a melatonin binding site in the song control system of the zebra finch. AB - In many avian species, singing is a circadian or seasonal behavior that appears to be widely dependent on gonadal steroid hormones. To explore the possibility of a further hormone-dependent vocal control mechanism driven by the action of melatonin, we examined the binding of iodinated melatonin (IMEL) in the vocal control network of adult and juvenile (22- and 40-day-old) zebra finches. IMEL binding areas of the zebra finch brain were localized and characterized by using quantitative in vitro autoradiography. In the vocal control system, dense IMEL binding sites were restricted to the nucleus hyperstriatalis ventrale, pars caudalis (HVC). The binding of IMEL to the HVC and to visual areas, e.g., the ectostriatum and the optic tectum, was saturable and showed a single class of high-affinity binding sites with binding affinities (Kds) in the range of 5-20 pM. Competition experiments with various indols and IMEL showed that the IMEL binding site in the zebra finch brain has properties similar to the high-affinity melatonin receptor described in the chicken, in the house sparrow, and in the mammalian brain and retina. Similar to the zebra finch HVC, the HVC of other songbirds, e.g., male canaries and male house sparrows, has the most intense IMEL binding of all areas of the vocal control network. The IMEL binding in the forebrain vocal control areas of the zebra finch, but not that in the visual processing areas, was sexually dimorphic in correlation with the sexually dimorphic neuroanatomy of the forebrain vocal control areas. In the HVC, there is a developmental increase in the maximal number of binding sites for IMEL and in the protein content, so that the adult phenotype of dense IMEL binding develops between day 40 and day 80. The distribution and developmental pattern of IMEL binding in the song system suggests that melatonin has a role in the motor control of singing. Melatonin binding sites in HVC could link HVC-based song control to circadian and circannual changes in the photoperiod independent of gonadal steroids. PMID- 8708013 TI - NMDA receptors and plasticity in adult primate somatosensory cortex. AB - Topographic maps in adult primate somatosensory cortex are capable of dramatic reorganizations after peripheral nerve injuries. In the present experiments, we have deprived a circumscribed portion of the hand map in somatosensory cortex of our adult squirrel monkeys by transecting the median nerve to one hand, and evaluated the hypothesis that N-methyl-d-aspartate (NMDA) glutamatergic receptors are necessary for the reorganization that follows within four weeks. In one monkey, we confirm previous results demonstrating that the deprived cortex has regained responsiveness in its expanse four weeks after median nerve transection. However, in three monkeys in which NMDA receptors were concurrently blocked, most of the deprived cortex remained unresponsive. Thus, much of the cortical "recovery" that typically follows peripheral nerve injury in adult monkeys is apparently dependent on NMDA receptors and may well be due to Hebbian-like changes in synaptic strength. Perhaps the elimination of the normally dominant inputs to "median nerve cortex" permits the gradual strengthening of correlations between the activity of the formally impotent presynaptic and deprived postsynaptic elements. These enhanced correlations may also have been made possible by reductions in intracortical inhibition as a necessary but not sufficient condition. PMID- 8708014 TI - Significance of the anti-Ro (SS-A) antibody in evaluation of patients with cutaneous manifestations of a connective tissue disease. AB - The anti-Ro(SS-A) antibody is arguably the most important antibody determination except for antinuclear antibodies in evaluation of patients suspected of having lupus erythematosus. During the past 25 years, studies have established the importance of this antibody in the evaluation of patients with atypical lupus erythematosus, who have a photosensitive dermatitis as the presenting sign. The purpose of this review is to demonstrate the utility and the necessity of this antibody determination in the evaluation of all patients with cutaneous manifestations who are suspected of having a connective tissue disease. PMID- 8708015 TI - Eosinophilic vasculitis in connective tissue disease. AB - BACKGROUND: Neutrophilic and lymphocytic vascular inflammation is common in vasculitis associated with connective tissue disease (CTD). We recently identified eight patients with CTD and eosinophilic vasculitis. OBJECTIVE: The purpose of this study was to characterize a variant form of vasculitis in CTD with eosinophilic infiltration. METHODS: Of 98 CTD patients with cutaneous necrotizing vasculitis, eight were found with predominantly eosinophilic vascular infiltration. Nine CTD patients with cutaneous neutrophilic vasculitis were identified for comparison. Clinical and laboratory findings were reviewed and compared. Indirect immunofluorescence for eosinophil granule major basic protein (MBP), neutrophil elastase, and mast cell tryptase was performed on lesional tissue. MBP levels and eosinophil survival enhancing activity were assayed in sera from three patients. RESULTS: The patients with eosinophilic vasculitis had depressed serum complement levels and peripheral blood eosinophilia; MBP levels were elevated in serum and eosinophil survival was prolonged. Immunofluorescence of tissue showed marked angiocentric eosinophil MBP staining with peripheral neutrophil elastase staining; mast cell tryptase staining was notably absent. The patients with neutrophilic vasculitis were variably hypocomplementemic and did not have peripheral blood eosinophilia. Immunofluorescence showed marked angiocentric neutrophil elastase staining with scattered eosinophil MBP staining; mast cell tryptase staining showed normal mast cell numbers. CONCLUSION: Patients with eosinophilic vasculitis, CTD, and hypocomplementemia show vessel wall destruction in association with vessel wall deposition of cytotoxic eosinophil granule MBP, which suggests that eosinophils mediate vascular damage in this disease process. In addition, perivascular mast cells appear diminished, thereby suggesting that mast cell degranulation occurs. PMID- 8708016 TI - The pattern of topical corticosteroid prescribing in the United States, 1989 1991. AB - BACKGROUND: Topical corticosteroids are widely used in the treatment of skin diseases. These preparations vary greatly in potency. They are available in both brand name and generic formulations, as well as in combination products. OBJECTIVE: Our purpose was to determine the pattern of topical corticosteroids prescribing in the United States and the relation of patient and prescriber attributes to the type of corticosteroid preparation prescribed. METHODS: Data from the 1989 to 1991 National Ambulatory Medical Care Survey were used to estimate the number of visits with a topical corticosteroid preparation prescribed and to identify prescribers with specific characteristics. RESULTS: In the United States, topical corticosteroids are prescribed or recommended at an average of 14 million visits per year to office-based health practitioners. Forty percent of these visits were to dermatologists. Dermatologists were 3.9 times more likely to prescribe very high potency steroids than were other physicians. Physicians other than dermatologists were 8.4 times more likely than dermatologists to prescribe combination agents containing moderate- or high potency topical corticosteroids and an antiinfective agent. CONCLUSION: The pattern of topical corticosteroid prescribing is substantially different for dermatologists and other physicians. These differences may reflect differences in severity or complexity of the disease or differences in prescribing habits. The importance of these differences to the outcome of treated patients is not established. PMID- 8708017 TI - Apoptosis is the mode of keratinocyte death in cutaneous graft-versus-host disease. AB - BACKGROUND: Satellite cell necrosis is a histopathologic hallmark of cutaneous graft-versus-host disease (GVHD) after allogeneic bone marrow transplantation. Although this interaction of lymphocytes with keratinocytes has features of immune cytolytic destruction, the details of the process are not known. OBJECTIVE: Our purpose was to investigate whether apoptosis is involved in the process of GVHD. METHODS: We used TdT-mediated dUTP-biotin nick end labeling (TUNEL) technique with biotinylated nucleotides. Formalin-fixed, paraffin embedded sections from skin biopsy specimens of patients with GVHD and normal skin were studied. RESULTS: Labeling of scattered individual keratinocytes in the epidermis and adnexal epithelium undergoing cytolytic degeneration and a proportion of the adjacent lymphocytes was noted in all specimens of GVHD. CONCLUSION: The positive staining of keratinocytes by the TUNEL method indicates that cell destruction in cutaneous GVHD involves apoptosis. PMID- 8708018 TI - The prevalence of hidradenitis suppurativa and its potential precursor lesions. AB - BACKGROUND: The morbidity of hidradenitis suppurativa can be considerable, but little is known about its epidemiology. OBJECTIVE: Our purpose was to describe the 1-year and point prevalences of hidradenitis suppurativa and its potential precursor lesions. METHODS: We obtained the histories and examined an unselected sample (599 persons) of the general population (1-year prevalence), and we performed physical examinations for a consecutive sample of 507 persons undergoing screening for sexually transmitted diseases (point prevalence). RESULTS: The point prevalence was 4.1% (95% confidence interval [CI] = 3.0-6.0) on the basis of objective findings. The 1-year prevalence of hidradenitis was 1.0% (CI = 0.4-2.2) on the basis of subject recollection only. The patients in the sample on which point prevalence is based were younger than those in the unselected sample of the general population (p < 0.001). Hidradenitis was significantly more common in women (p = 0.037), which may result from a female preponderance of genitofemoral lesions (odds ratio [OR] = 5.4; CI = 1.5 - 19.3). No sex difference was found in the prevalence of axillary lesions. CONCLUSION: Hidradenitis suppurativa is significantly more common than hitherto estimated. The female preponderance of patients is confirmed, except for patients with axillary lesions. Additional longitudinal studies are necessary to assess the importance of potential precursor lesions such as noninflamed nodules or comedones. PMID- 8708019 TI - Hepatitis C virus infection in patients with urticaria. AB - BACKGROUND: Hepatitis C virus (HCV) infection induces variable skin manifestations. OBJECTIVE: Our purpose was to determine whether there is an association between HCV infection and urticaria. METHODS: Antibody to HCV (anti HCV) and HCV genotypes were determined in patients with urticaria and in a control population. RESULTS: Anti-HCV was detected in 19 (24%) of 79 patients with urticaria, and HCV RNA was detected in 17 (22%). Genotypes of HCV were II/1b in 12 (71%), III/2a in 4 (24%), and IV/2b in 1 (6%). The 17 patients with HCV RNA were older (53 +/- 14 vs 41 +/- 14 years, p < 0.01), and their eruption lasted longer (35% vs 6%, p < 0.05) and left pigmentation more frequently (53% vs 3%, p < 0.001). They had higher levels of alanine aminotransferase (67 +/- 34 vs 25 +/- 17 U/L, p < 0.001), aspartate aminotransferase (51 +/- 23 vs 21 +/- 8 U/L, p < 0.001), zinc turbidity test (12.8 +/- 3.1 vs 9.3 +/- 3.7 Kunkel units, p < 0.001), and IgG (1919 +/- 320 vs 1622 +/- 349 mg/100 ml, p < 0.01) than the patients without HCV RNA. CONCLUSION: HCV could be a significant cause of urticaria. Chronic urticaria associated with HCV infection has peculiar clinical, serologic, and biochemical characteristics that could make it a distinct clinical entity with an indication for interferon therapy. PMID- 8708020 TI - Long-term results of topical immunotherapy in children with alopecia totalis or alopecia universalis. AB - BACKGROUND: Topical immunotherapy has been used in the treatment of children with alopecia areata with encouraging results. OBJECTIVE: Our purpose was to determine the long-term results in 33 children with severe alopecia areata treated with topical immunotherapy. METHODS: From 1983 to 1989 we treated 33 children with topical immunotherapy with squaric acid dibutylester. RESULTS: Complete hair regrowth was observed in 10 children (30.3%). During the follow-up period (mean, 5.9 years; range, 4 to 12 years), 7 of these 10 patients had severe relapses that were not responsive to further treatment. Only three clinically benefited from topical immunotherapy. Two maintained complete hair regrowth after treatment was stopped. CONCLUSION: Our results indicate that only a small proportion of children with severe alopecia areata will obtain a persistent benefit from topical immunotherapy. PMID- 8708021 TI - Improvement of monitoring of melanocytic skin lesions with the use of a computerized acquisition and surveillance unit with a skin surface microscopic television camera. AB - BACKGROUND: Photographic documentation of melanocytic skin lesions is important. Storage and retrieval of slides, however, take much time and space. OBJECTIVE: Our purpose was to develop and clinically test a computerized acquisition and surveillance (CAS) unit with a television camera for monitoring including measurements of lesional areas. METHODS: A CAS unit connected with a skin surface microscopic television camera was used for monitoring of melanocytic nevi (MN). The lesional area and the skin surface microscopic appearance (SMA) were analyzed after 10 to 21 months in 54 of 1355 MN. RESULTS: In 19 MN (35.2%), changes were found. In eight cases, changes in size of more or less than 15% were detected; in five cases only the SMA changed. In six cases both characteristics changed. CONCLUSION: In approximately 25% of MN, changes were only detectable in the SMA but not with area measurements. This favors the use of systems such as CAS because only they allow a time-saving comparison of actual and previous images. PMID- 8708022 TI - Treatment of psoriasis with polyethylene sheet bath PUVA. AB - BACKGROUND: Bath PUVA has been shown to be an effective alternative treatment for psoriasis with fewer systemic side effects than oral methoxsalen (8-MOP). The cost of 8-MOP and the need for a bath unit have prevented wider use of this treatment. OBJECTIVE: We investigated the safety and efficacy of sheet bath PUVA by restricting the volume of the psoralen/bath water solution to 10 L with the aid of a polyethylene sheet. METHODS: Fifty-eight patients with chronic plaque type psoriasis were treated with bath PUVA in a concentration of 0.5 mg of 8-MOP per liter of water. RESULTS: The group required a median of 17 baths (95% confidence interval [CI], 14-20) for clearance. Total UVA dose for the entire group was 26 J/cm2(95% CI, 18-47). CONCLUSION: Sheet bath PUVA is safe, efficient, and easy. This regimen can significantly reduce the amount of 8-MOP required, thereby resulting in a favorable cost/benefit ratio. PMID- 8708023 TI - Dye-enhanced diode laser photothermal ablation of skin. AB - BACKGROUND: Near-infrared diode lasers are promising new light sources. However, their applications for rapid and precise removal of tissue is limited because their radiation is poorly absorbed by most tissue. OBJECTIVE: Our purpose was to determine the interaction of diode laser radiation with tissue by topically applying absorbing dye to enhance laser-tissue interaction and to examine the ability of this approach to create superficial and localized thermal effects in skin. METHODS: Immediate and long-term responses of guinea pig skin to low-power (3 to 5 W X 1 sec) diode laser (805 nm) irradiation in conjunction with indocyanine green as a dye were studied. Tissue response was characterized on the basis of gross and histopathologic analysis of biopsy specimens taken at different times. RESULTS: The observed depths of thermal damage with dye were greater than those produced by diode laser used alone at higher power settings (10 W X 1 sec). CONCLUSION: Interaction of diode laser radiation enhanced by topical application of dye causes localized superficial thermal effects in skin. This may offer a new modality in the treatment of superficial lesions. PMID- 8708024 TI - Intermittent fluconazole dosing in patients with onychomycosis: results of a pilot study. AB - BACKGROUND: Therapeutic limitations of griseofulvin in treating onychomycosis have led to a search for alternative antimycotic agents. An optimal dosing regimen for fluconazole has yet to be defined. OBJECTIVE: Our purpose was to evaluate the intermittent use of fluconazole (either once-weekly or alternate-day dosing) without concurrent nail avulsion in patients with moderate to severe onychomycosis. METHODS: Eleven patients with mycologically confirmed onychomycosis of the toenails or fingernails (43 infected nails) were treated with intermittent fluconazole until clinical cure was obtained. Eight patients received fluconazole 300 mg once weekly, one patient received 200 mg once weekly, and two patients received 100 or 200 mg of fluconazole every other day. Eight patients also used an adjunctive topical antimycotic preparation. RESULTS: All six patients with toenails involved (32 infected nails) were clinically cured after a mean treatment duration of 6 months, and all five patients with fingernails involved (11 infected nails) were cured after 3.7 months. There were no significant clinical or laboratory adverse events. CONCLUSION: Intermittent fluconazole, taken once weekly or on alternate days, is a well-tolerated and efficacious method to treat onychomycosis. PMID- 8708025 TI - Transverse scalp sections: a proposed method for laboratory processing. AB - BACKGROUND: Transverse sections of human scalp biopsy specimens can provide both qualitative and quantitative information about follicular histopathology not readily available in vertically sectioned specimens. OBJECTIVE: We propose a method for making the processing and interpretation of such specimens easier. METHODS: All scalp biopsy specimens obtained during the past 18 months in our institution for the evaluation of alopecia were processed for transverse sections by means of a technique employing trisection or quadrisection (rather than the standard bisection), and maintaining all sections in the same anatomic orientation (deep to superficial) in all tissue pieces on microscopic slides. RESULTS: More than 120 transversely sectioned specimens from more than 75 patients have been processed with this technique. The typical number of slides cut per specimen decreased from between 12 and 20 to between 1 and 4, with no loss of clinical information. CONCLUSION: This technique allows transverse sections for evaluation of alopecia to be processed in a more cost-effective manner and compares favorably with previously published techniques in providing diagnostic information. PMID- 8708026 TI - Premalignant keratinocytic neoplasms. AB - Premalignant keratinocytic keratoses are common, especially in pale-complected persons in whom they appear most often as an actinic keratosis. Although the actinic keratosis has a very low malignant potential, arsenic, tar, thermal, scar, reactional, and radiation keratoses may be more clinically aggressive. This article discusses these premalignant keratinocytic neoplasms. PMID- 8708027 TI - Surgical pearl: suturing--control and conservation. PMID- 8708028 TI - Hypoallergenicity and the dermatologist's perception. PMID- 8708029 TI - International Foundation for Dermatology: the first 8 years. PMID- 8708030 TI - Comparison of the irritancy of hand dishwashing liquids with modified patch testing methods. PMID- 8708031 TI - Elusive migratory subcutaneous dirofilariasis. PMID- 8708032 TI - Atypical fibroxanthoma in a renal graft recipient. PMID- 8708033 TI - Delayed onset of skin fibrosis after the ingestion of eosinophilia-myalgia syndrome-associated L-tryptophan. PMID- 8708034 TI - Recalcitrant giant molluscum contagiosum in a patient with AIDS. PMID- 8708035 TI - A multicenter trial of calcipotriene ointment and halobetasol ointment compared with either agent alone for the treatment of psoriasis. PMID- 8708036 TI - Interferon alfa-induced interstitial pneumonitis in a patient with cutaneous T cell lymphoma. PMID- 8708037 TI - Cimetidine therapy for multiple viral warts in children. PMID- 8708038 TI - Oral calcitriol (1,25-dihydroxyvitamin D3) does not augment UVB phototherapy for plaque psoriasis. PMID- 8708039 TI - Hydroxyurea-induced nail pigmentation. PMID- 8708040 TI - Association of glaucoma with cutis marmorata telangiectatica congenita: a localized anatomic malformation. PMID- 8708041 TI - A centimeter here, a centimeter there. PMID- 8708042 TI - A centimeter there, a centimeter there: does it matter? PMID- 8708043 TI - Pancreatic panniculitis. PMID- 8708044 TI - Occlusive treatment in psoriasis: how does it work? PMID- 8708045 TI - Thrombocytopenia caused by fluconazole. PMID- 8708047 TI - CT-guided intraarticular corticosteroid injection into the sacroiliac joints in patients with spondyloarthropathy: indication and follow-up with contrast enhanced MRI. AB - PURPOSE: Our goal was to prospectively study the therapeutic efficacy of CT guided intraarticular corticosteroid instillation of inflamed sacroiliac joints (SIJs) in patients with spondyloarthropathies and to evaluate the role of MRI as a procedure for establishing the indication and for therapeutic follow-up. METHOD: A total of 103 CT-guided corticosteroid injections of the SIJs were performed in 66 patients with inflammatory back pain (IBP): 37 bilateral, 29 unilateral. All patients did not respond to an immediately preceding 4 week nonsteroidal antiinflammatory therapy. Forty milligrams of a crystalline long acting corticoid was instilled in each joint. All 66 patients underwent continuous clinical follow-up at 10 to 12 week intervals after corticosteroid injection to a maximum of 18 months. The degree of subjective complaints before and after the intervention was recorded using a analogue scale (0 = no pain, 10 = unbearable pain). Dynamic contrast-enhanced (Gd-DTPA, 0.1 mmol/kg body wt) MRI with quantitative determination of contrast enhancement was performed in all patients before the therapeutic intervention and in 38 patients at 8 +/- 4 months. RESULTS: Sixty-one of the 66 study patients (92.5%) showed a statistically significant abatement of subjective complaints from 8.8 +/- 1.3 to 3.3 +/- 2.3 (p < 0.01) at 1.7 +/- 1.1 weeks after intervention, and this improvement lasted for 10 +/- 5 months. The percentage contrast enhancement at dynamic MRI likewise showed a significant reduction from 100.3 +/- 48% before to 44.3 +/- 36.1% after intraarticular cortisone administration (p < 0.01). CONCLUSION: CT-guided intraarticular corticosteroid instillation in the SIJs may be regarded as an effective therapy for florid sacroiliitis. The severity of inflammation and the response to therapy can be determined quantitatively by dynamic MRI. PMID- 8708048 TI - Kinematic MRI of the knee using a specially designed positioning device. AB - To evaluate the benefits of a specially designed positioning device, kinematic MR studies of the patellofemoral joint and the cruciate ligaments were performed. Handling of the device and positioning of the patient's leg were easy and took only a few minutes. The positioning device was helpful for patients to guide their joint motion and to obtain sufficient image quality for kinematic MR studies of both cruciate ligaments and the patellar tracking. PMID- 8708049 TI - Preoperative CT localization and marking of osteoid osteoma: description of a new technique. AB - We describe a new technique for the preoperative localization of an osteoid osteoma nidus. Four lesions (mean patient age 26 years) were localized with CT. A needle was advanced into the cortex, methylene blue was injected, and a self retaining localization wire was then left in place. A final CT image confirmed localization. The site of marking was identified intraoperatively by following the guide wire to the cortical mark, permitting nidus excision with minimal resection of bone. PMID- 8708046 TI - Madelung deformity in skeletally immature patients: morphologic assessment using radiography, CT, and MRI. AB - PURPOSE: The purpose of this study was to define the pathoanatomy of the distal radius and surrounding soft tissues, identify the factors that may contribute to diminished forearm rotation, and relate these findings to alterations in wrist motion in skeletally immature patients with the Madelung deformity. METHOD: Four skeletally immature female patients with bilateral Madelung deformities (eight wrists) underwent evaluation of each wrist with radiography, CT, and MRI to assess the morphology of the deformity. Two patients (four wrists) had isolated idiopathic Madelung deformities, and two patients (four wrists) had Madelung deformities secondary to dyschondrosteosis. RESULTS: Radiographically, all wrists demonstrated dorsal bowing of the radius, marked ulnar tilting of the radius and radial tilting of the ulna, volar tilting of the distal articular surface of the radius, and triangulation of the epiphysis. On CT, patients with the idiopathic deformity demonstrated dorsal ulnar subluxation and relative supination of the carpus with respect to the distal radius. Patients with dyschondrosteosis demonstrated no ulnar subluxation and relative pronation of the carpus. All wrists demonstrated a fixed pronated deformity of the distal radius. On MRI, a physeal bar that bridged the distal metaphysis of the radius to the epiphysis was identified in all eight wrists, located on the volar aspect of the radius at the lunate facet. An anomalous volar ligament, a volar radiotriquetral ligament, and the short radiolunate ligament were hypertrophied in seven wrists. CONCLUSION: Based on its location, it is likely that a physeal bar impedes the normal development of the distal radius ulnarly. Hypertrophy of the short radiolunate ligament may be an important contributing factor to carpal pyramidalization owing to tethering on the volar pole of the lunate. Diminished forearm rotation is likely related to carpal malalignment, a fixed pronated deformity of the distal radius, and dorsal bowing of the radius. PMID- 8708050 TI - Imaging of "iliac horns" in nail-patella syndrome. AB - Hereditary osteo-onychodysplasia, also known as the nail-patella syndrome (NPS), consists of a clinical tetrad of nail dysplasia, hypoplastic or absent patella, radial head dislocation, and iliac horns. Bilateral posterior iliac horns are observed only in NPS and present in >80% of patients. We describe the appearance, location, and structure of iliac horns by CT, MRI, and 3-dimensional CT reconstruction. PMID- 8708051 TI - Cerebral blood volume maps with dynamic contrast-enhanced T1-weighted FLASH imaging: normal values and preliminary clinical results. AB - PURPOSE: In this article we investigate the application of a method that uses the relaxation effect of bolus-like injected Gd-DTPA to quantify regional cerebral blood volume (rCBV). The aim of the study was to determine if the method provides correct rCBV values in healthy subjects as well as to obtain additional diagnostic information for patients with a glioma or stroke. METHOD: Twenty healthy subjects, 12 patients with brain infarctions, and 18 patients with gliomas were examined. A series of 64 sequential images of one slice was recorded during bolus transit with a FLASH sequence. The measured signal intensity-time curves were converted pixel-wise to concentration-time curves from which the rCBV images were calculated applying the indicator dilution method. RESULTS: An average value for gray and white matter of 4.4 +/- 1.6 vol% was obtained for the group of healthy subjects. The grading of the tumors could be classified according to the differences of their corresponding rCBV values. Fifty percent of the infarct patients had to be excluded from the analysis in the acute phase due to mispositioning of the slice and data degradation by gross motion artifact. Different rCBV values were found for areas that develop later into gliotic scars or cystic necrosis. CONCLUSION: The proposed method is easy to apply in clinical routine MR investigations and provides valuable information for noninvasive, preoperative assessment of tumor grading. It can also provide additional criteria for estimating the histological outcome and with it the degree of ischemia in stroke patients. PMID- 8708053 TI - Chordomas of the skull base: dynamic MRI. AB - OBJECTIVE: To investigate the role of dynamic MRI in the diagnosis of chordomas at the skull base. MATERIALS AND METHODS: Six patients with surgically proven chordomas of the skull base were examined with spin-echo technique (repetition time/echo time 200 ms/15 ms). Serial imaging was performed every 30 s for 240 s after a rapid injection of gadopenetate dimeglumine (0.1 mmol/kg). The contrast material enhancement ratio (CER) was analyzed. The CER was obtained for each region of interest as follows (Sn - So) x 100/Smax - So, where Sn indicates signal intensity at each dynamic phase; So, signal intensity on the image obtained before administration of contrast material; and Smax, maximum signal intensity of all dynamic imaging. RESULTS: All chordomas showed very slow but steady increase of contrast enhancement with no peak within 240 s. The CERs of six cases at 30 s are 13 to 40 (mean value +/- standard deviation: 22.8 +/- 10.2). CONCLUSION: Dynamic MRI may be helpful in differentiating between chordomas and other tumors of the skull base. PMID- 8708052 TI - Magnetization transfer imaging of diffuse axonal injury following experimental brain injury in the pig: characterization by magnetization transfer ratio with histopathologic correlation. AB - PURPOSE: Our goal was to evaluate the use of the magnetization transfer ratio (MTR) in the detection of diffuse axonal injury (DAI) resulting from traumatic brain injury in a swine model. METHOD: DAI was created by applying a nonimpact, coronal plane, rotational acceleration to the heads of miniature swine (n = 4). GE imaging was performed with and without off-resonance MT saturation. Histologic correlation of axonal injury with MRI was performed 7 days postinjury. Thirty-one subcortical white matter regions and 10 deep white matter regions were selected for the direct comparison of histologic data and MTR measurements. RESULTS: Nineteen of 41 examined locations exhibited histologic evidence of axonal injury. The mean MTR in regions with axonal damage was significantly less than in regions without axonal damage. These changes were observed both in regions demonstrating high signal intensity on T2-weighted images (T2WI) (p <0.0001, n = 6) and in regions with no signal intensity change on T2WI (p < 0.05, n = 13). CONCLUSION: These results suggest that the measurement of MTR may have the potential for evaluation axonal damage in DAI following traumatic brain injury even when conventional T2WI does not demonstrate the lesion. PMID- 8708054 TI - MR findings in Tay-Sachs disease. AB - OBJECTIVE: MR and CT findings in two patients of Tay-Sachs disease are reported. MATERIALS AND METHODS: In two patients of Tay-Sachs disease, MR and CT studies were conducted in the early phase (11 and 10 months of age, respectively) and one was followed-up with MR in the late phase at the age of 5 years and 8 months. Six MR studies on this disease (including 3 other cases documented in the literature) were reviewed and correlated with the pathological phases of this disease. RESULTS: MRI findings drastically changed during the phases of disease process in one case. In the early phase, T2-weighted MR images demonstrated hyperintensity in both basal ganglia and thalamus. MR images obtained in the late phase demonstrated markedly advanced brain atrophy and diffuse white matter lesions, which were hyperintense on the T2-weighted images. T1-weighted images exhibited hyperintensity in the basal ganglia, thalamus, and along the cortical layer of the cerebrum, which appeared to by hypointense on T2-weighted images. CONCLUSION: MR was apparently superior to CT in clearly delineating lesions of this disease. These abnormal signal intensities in the late phase may reflect the pathological findings, marked accumulation of GM2 ganglioside in the basal ganglia and thalamus and along the cerebral gray matter. PMID- 8708055 TI - Intracranial chondroma arising from the cranial vault: CT and MR appearance. AB - A case of intracranial chondroma arising from the right parietal bone in a 37 year-old woman is presented. CT and MRI findings as well as the differential diagnosis of the lesion are discussed. PMID- 8708056 TI - Venous air emboli identified on head and neck CT scans. AB - OBJECTIVE: Small venous air emboli probably occur frequently. Our purpose was to describe the locations of small venous air emboli detected on CT scans of the head and neck and their clinical presentations. MATERIALS AND METHODS: The head CT scans of 17 patients and neck CT scans of 10 patients with suspected venous air emboli were reviewed and the locations of the gas collections were recorded. The charts of these patients were reviewed to identify possible sources of these gas collections and any symptoms they may have produced. RESULTS: The most likely source of these gas collections was venous air emboli. The neck CT scans demonstrated gas in the inferior internal jugular vein, subclavian vein, and small anterior neck veins. The head CT scans demonstrated gas in the cavernous sinus, the frontal and temporal scalp, the infratemporal fossa, the carotid canal, the straight sinus, the superior ophthalmic vein, the superior sagittal sinus, extracranially in the region of the foramen magnum, and in a canal in the skull base for an emissary vein or the inferior petrosal sinus. Most of the intravenous lines were placed within 6 h of the scans demonstrating gas. The gas collections did not produce symptoms. CONCLUSION: Asymptomatic venous air emboli can be identified in several locations in the head and neck. The time between manipulation of intravenous lines and the scan, the position of the patient, and the anatomy of the patient probably all affect the likelihood of identifying venous air emboli on CT scans. PMID- 8708057 TI - Nasopharyngeal carcinoma with intracranial spread: CT and MR characteristics. AB - PURPOSE: Nasopharyngeal carcinoma (NPC) frequently spreads intracranially. We compare CT and MRI in identifying intracranial spread and reexamine the route of infiltration. METHOD: One hundred fourteen consecutive patients with proven NPC were evaluated prospectively with T1-, T2-weighted, contrast-enhanced MRI and CT. RESULTS: MRI showed 35 (31%) patients with middle cranial fossa involvement. Twenty-nine (25%) patients had cavernous sinus infiltration, while six (5%) showed only dural thickening. The most common route of spread is through the foramen ovale (FO) (12/35 patients, 34%), followed by skull base destruction (6/35 patients, 17%), foramen lacerum (FL) (6/35 patients, 17%), sphenoid sinus (6/35 patients, 17%), and combined FO and FL (5/35 patients, 14%). Using MRI as a standard, CT demonstrated the following involvement: cavernous sinus in 26 of 29 (90%) patients, FO in 9 of 12 patients, skull base in 6 of 6 patients, FO and FL in 3 of 5 patients, FL in 6 of 6 patients, sphenoid sinus in 6 of 6 patients and dura in 0 of 18 patients. CONCLUSION: It is believed that NPC most commonly spreads intracranially via the FL or by direct erosion. Perineural spread through the FO is an important route, which explains why with CT evidence of cavernous sinus involvement there may be no skull base erosion. These findings are best seen on MRI. PMID- 8708058 TI - CT appearance of microcystic meningioma. AB - Microcystic meningioma is an unusual variant, which has recently been proposed for inclusion in the WHO Classification of Central Nervous System Tumors. Its unique structure produces findings that may be confusing to radiologist and pathologist alike. A case is reported and the English language literature reviewed. PMID- 8708059 TI - MRI of dumbbell-shaped spinal tumors. AB - PURPOSE: The purpose of our study is to review the clinical features and MR findings in patients with dumbbell-shaped spinal tumors that affect the intervertebral foramen. METHOD: Two radiologists retrospectively evaluated a total of 19 tumors from the period 1989-1994 that exhibited dumbbell configuration. These included 11 benign tumors, including 9 neurogenic ones, and 8 malignant tumors. RESULTS: The time between onset of symptoms and admission was shorter in patients with malignant tumors (mean 3.9 months) than in patients with benign tumors (mean 41.1 months). There was a statistically significant difference (p < 0.05) in the short/long axis ratios (minimum tumor diameter divided by maximum tumor diameter) in the vertebral canal components between benign and malignant tumors. All malignant tumors in this study were extradural and paraspinal type (p < 0.05). Most benign tumors had regular margins and enlarged intervertebral foramina, while most malignant ones had irregular margins (p < 0.05). Many neurogenic tumors had regular margins and enlarged intervertebral foramina, while most malignant ones had irregular margins (p < 0.05> Many neurogenic tumors had cystic lesions (77.8%) and a string-of-beads structure (44.4%). Those tumors that surrounded the thecal sac in the vertebral canal and whose adjacent vertebral bodies had abnormal signal intensities were infiltrating and malignant. CONCLUSION: There are many kinds of dumbbell-shaped spinal tumors other than neurogenic ones. MRI is useful in diagnosing dumbbell shaped spinal tumors, especially those of neurogenic origin, and aids in distinguishing benign from malignant tumors. PMID- 8708060 TI - MRI of unfused lumbar segments after spondylodesis. AB - PURPOSE: Our goal was to find a causal relation between disk degeneration in nonfused lumbar segments, back pain, and the angle of scoliotic curves 21-30 years after dorsal spondylodesis. METHOD: Thirty-two patients with scoliosis had been treated by dorsal spondylodesis 21-30 years prior to this study (mean 25.2 +/- 2.4 years). With use of MRI, signs of disk degeneration in the nonfused lumbar segments (n = 131, mean 4.1 +/- 1.4) were evaluated. Back pain evaluation was performed using the Oswestry disability score. On plain X-ray films, the angle of scoliotic curves was measured (Cobb technique). Percentage and absolute number of degenerated disks were compared to the fusion level (number of unfused caudal segments), the Oswestry score, and the Cobb angle. RESULTS: Disk degeneration was present in 20 of 32 patients (62.5%) and in 37 of 131 unfused lumbar segments (28.2%). Of eight patients (24.2%), seven had moderate back pain and one severe back pain. The mean lumbar scoliotic curve was 44.4 +/- 24.9 degrees. No statistically significant correlation was found between patients with disk degeneration and fusion level, amount of back pain, or lumbar scoliotic angle (p < 0.05). CONCLUSION: A lack of correlation between disk degeneration in unfused lumbar segments and other parameters like fusion level, back pain, and scoliotic angle can be assessed in the long-term follow-up of multisegment fusion of scoliotic curves. PMID- 8708061 TI - A simple noninvasive stereotactic device for routine MR head examinations. AB - A simple device to define reproducible reference points in MR examinations was designed. The device can be worn like a pair of eyeglasses and has reference structure visible in MR images. Combined with a specially designed algorithm, the imaging plane can be aligned to a predefined coordinate system in one step, using a single slice through the device. The device can be produced at low cost and is suitable for routine examinations as well as surgical planning. PMID- 8708062 TI - Unifocal primary amyloidoma of the spine causing compression of the cervical spinal cord: MR findings. PMID- 8708063 TI - Tree-in-bud pattern: frequency and significance on thin section CT. AB - PURPOSE: Our goal was to describe those diseases of the airways that manifest the tree-in-bud (TIB) pattern on CT scan and to establish a differential diagnosis for this CT scan finding. METHOD: We prospectively collected cases with the TIB pattern on CT and reviewed the scans of patients with histories pertaining to small airway disease. CT scans were performed at 1 to 3 mm collimation. RESULTS: Twenty-six of 27 cases with the TIB pattern had associated bronchiectasis or proximal airway wall thickening. One case with normal proximal airways had an acute aspiration. In addition, we reviewed 141 scans of patients with emphysema, respiratory bronchiolitis (RB), bronchiolitis obliterans (BO), bronchiolitis obliterans organizing pneumonia (BOOP), extrinsic allergic alveolitis (EAA), bronchiectasis, bronchitis, and pneumonia. Of the CT scans with bronchiectasis, 25.6% had TIB, and 17.6% of CT scans with acute infectious bronchitis or pneumonia had this pattern. None of the patients with emphysema, BO, BOOP, EAA, or RB had this pattern. CONCLUSION: The TIB pattern on CT scan is mostly associated with pulmonary infections that commonly involve the large airways. This pattern was present in 17.6% of cases with acute bronchitis or pneumonia and 25.6% of cases with bronchiectasis. PMID- 8708064 TI - High resolution CT findings in community-acquired pneumonia. AB - PURPOSE: Our goal was to clarify the high resolution CT (HRCT) findings of community-acquired pneumonia based on pathologic findings and to make a differential diagnosis between bacterial and atypical pneumonias. METHOD: This study evaluated 32 cases with community-acquired pneumonia, including 18 cases with bacterial pneumonia and 14 cases with atypical pneumonia [mycoplasma pneumonia (n = 12), chlamydia pneumonia (n = 1), and influenza viral pneumonia (n = 1)]. HRCT images in these cases were space consolidation, ground-glass attenuation, thickening of the bronchovascular bundle, and distribution of abnormal attenuation. RESULTS: Bacterial pneumonia frequently showed air space consolidation with segmental distribution (72.2%) that tended to locate at the middle and outer zones of the lung. Atypical pneumonia frequently showed centrilobular shadow (64.3%), acinar shadow (71.4%), air space consolidation and ground-glass attenuation with lobular distribution (57.1 and 85.7%, respectively), and tendency of the lesions to distribute at the inner layer of the lung in addition to the middle and outer layers (85.7%). CONCLUSION: Characteristic HRCT findings of both bacterial and atypical pneumonia were demonstrated. These HRCT features seemed to reflect pathologic findings and the manner of lesional progression. This information may support the appropriate antibiotic therapy in medical practice. PMID- 8708065 TI - Weil disease: importance of imaging findings for early diagnosis. AB - Weil disease is a rare and severe outcome of infection with leptospires. We report the radiographic appearance of this disease in a patient with hepatic, renal, and pulmonary involvement. Imaging findings added significantly to an early correct diagnosis and, thus, to immediate and successful therapy in this critically ill patient. PMID- 8708066 TI - MR visualization of aortic valve vegetations. AB - PURPOSE: The purpose of this report is to describe the MR findings of aortic vegetations in two patients. METHOD: MR imaging using SE and fast GE (Fastcard) sequences was performed in the axial and coronal planes. RESULTS: The vegetations were not visible with SE MRI, but were clearly visible with fast GE imaging (Fastcard). The vegetations appeared as areas of low signal at valve leaflets in contrast to the bright flowing blood. CONCLUSION: MR evaluation in patients with infective endocarditis is useful in clarifying echocardiographic findings and establishing the diagnosis in previously undiagnosed patients. PMID- 8708067 TI - Differential density of normal renal parenchyma on nonenhanced CT: a new observation. AB - PURPOSE: Nonenhanced CT scans through the kidneys commonly show a difference in radiopacity between the inner peripelvic portion of the renal parenchyma and the more peripheral regions of the renal parenchyma. This normal observation has not been described. METHOD: We reviewed 50 nonenhanced CT scans. Renal parenchyma was evaluated by both visual inspection and density measurements of inner and outer aspects of the kidneys in 38 and visual inspection only in 12. A narrower window was used to better show density differences (100 W, 35 L). We recorded the indication for the CT, history, and recent blood chemistries. RESULTS: The inner parenchyma was of greater radiopacity than the outer parenchyma in 35 of 50 cases by visual inspection and in all densitometry cases (38 of 38). There was no correlation with clinical history or blood chemistry. CONCLUSION: A difference in density between the inner and outer portions of the renal parenchyma is very common and will often be seen if one looks for it. We have not shown an etiology for this difference, and indeed, it may be physiological. One must be aware of this finding to avoid mistaking it for pathology. PMID- 8708068 TI - CT urography in the evaluation of urinary tract disease. AB - OBJECTIVE: The purpose of this study was to evaluate the efficacy of using limited CT through the kidneys following excretory urography in selected situations. MATERIALS AND METHODS: Within 2 h of excretory urography 568 patients had add-on CT without using additional contrast medium. In 487 patients with hematuria the results of the combined studies were compared with blinded prospective interpretations of each. In addition 81 patients had add-on CT for specific urographic questions. RESULTS: Of the 345 renal parenchymal masses seen on CT in both groups, only 185 were detected prospectively on excretory urography. For the 81 patients who had CT because of abnormalities seen on excretory urography, including 54 with pseudotumors, findings were clarified and the correct diagnosis made. In 27 of 30 patients with lesions in the collecting system the lesions were detected only on excretory urography. CONCLUSION: Excretory urography followed by limited CT increases the diagnostic accuracy for detecting, characterizing, or ruling out urinary tract disease by combining the high sensitivity of traditional excretory urography for detecting lesions in the collecting system and ureter with the high accuracy of CT for parenchymal and perinephric lesions without an additional patient visit or additional contrast medium. Because it leaves so few unanswered questions, the combination of standard excretory urography and limited CT is an excellent screening study in selected situations. It can be performed quickly, at controlled cost, is highly accurate, and is convenient for the patient, the referring physician, and the examining department. PMID- 8708069 TI - MR cholangiography in biliary obstruction using half-Fourier acquisition. AB - PURPOSE: Our goal was to evaluate biliary obstruction using a T2-weighted, turbo, SE MR sequence with half-Fourier acquisition (HASTE). METHOD: A prospective evaluation of 21 consecutive patients with clinical evidence of obstructive jaundice was carried out comparing HASTE MR cholangiography (MRC) to endoscopic retrograde cholangiopancreatography and percutaneous transhepatic cholangiography. A control group of five normal volunteers was also evaluated. The study group was imaged with a 1.5 T MR scanner using a body coil. The HASTE sequence was applied in axial, coronal, and oblique sagittal planes. Ultrafast acquisition scanning times allowed the use of a single breath hold. Bile duct dilatation, level of obstruction, and cause of obstruction were assessed on both imaging modalities by two radiologists blinded to the clinical diagnosis and to each other's results. RESULTS: All studies were interpretable with anatomy well seen in 82% of the cases. MRCs of a normal control group were correctly interpreted. The presence of biliary dilatation was accurately depicted by HASTE MRC in 100% of patients with complete interobserver agreement. The level of obstruction was depicted correctly in 87% of patients with 93% interobserver agreement. The right main duct was seen by MRC in 80% of obstructed systems. The left main duct was seen in all obstructed patients. The gallbladder was identified in 88% of patients by MRC. Common bile duct stricture and stones could be differentiated as a cause of obstruction in all cases. CONCLUSION: MRC using the HASTE imaging sequence can safely and accurately depict the presence and level of biliary obstruction. The fast acquisition time of 13 s/scan makes the technique suitable for uncooperative and ill patients. HASTE MRC should be considered an alternative procedure to direct cholangiography in selected patients. PMID- 8708070 TI - Contrast-enhanced spiral CT of the liver: effect of injection time on time to peak hepatic enhancement. AB - PURPOSE: On contrast-enhanced hepatic CT, maximum tumor detection of liver metastases from colorectal cancer is achieved at peak hepatic enhancement. We investigated the relationship between injection time of contrast medium and time to peak hepatic enhancement (TPHE) after the end of injection. METHOD: One hundred nineteen patients without a cardiovascular disorder were enrolled in this study. Before the spiral CT was performed, a small amount of contrast medium was injected and a single level dynamic CT was performed to evaluate aortic enhancement and to determine the scan start time of the spiral examination. Patients were divided into three groups; contrast medium was injected over 30 s in 40 patients (Group A), 45 s in 39 patients (Group B), and 60 s in 40 patients (Group C). The TPHE after the end of injection was measured, and the difference among the groups was compared using one-way analysis of variance. A p value of <0.05 was considered a statistically significant difference. RESULTS: The TPHEs of the groups were 25.3 +/- 4.6 s (Group A), 27.0 +/- 5.8 s (Group B), and 24.4 +/- 4.6 s (Group C) and were similar in value. No statistically significant difference was observed (p = 0.067). CONCLUSION: Hepatic enhancement reaches its peak at approximately 25 s after the end of contrast medium injection irrespective of injection time. PMID- 8708071 TI - Intraperitoneal drop metastases from hepatocellular carcinoma: CT and angiographic findings. AB - Our goal was to describe the radiological manifestations of intraperitoneal drop metastases (IPDMs) from hepatocellular carcinoma (HCC). METHODS: Seven patients with clinically (n = 4) or pathologically (n = 3) proven IPDMs from HCC were retrospectively analyzed. Medical records, CT scans (n = 7), and angiographic findings (n = 5) were reviewed. RESULTS: On CT scans, all seven patients had single or multiple nodular masses in the liver. In six patients, the hepatic masses showed an exophytic growth. Single (n = 4) or multiple (n = 3) discrete omental masses, 3-10 cm in diameter, were also demonstrated in all patients. Intratumoral necrosis in the omental mass and engorgement of adjacent omental vessels were demonstrated in five and three patients, respectively. On angiography, the omental masses showed hypervascularity supplied by omental branches (n = 4) and superior mesenteric (n = 2) and inferior mesenteric (n = 1) artery and also showed prominent draining veins. CONCLUSION: The radiological manifestations of IPDM from HCC are single or multiple discrete hypervascular masses in the omentum. These are quite different from those of usually carcinomatosis peritonei. PMID- 8708072 TI - Abdominal macronodular tuberculomas: MR findings. AB - PURPOSE: Our goal was to determine the appearance of abdominal macronodular tuberculomas on MRI. METHOD: MR findings of two patients with abdominal macronodular tuberculoma, one of the liver and the other in the kidney, were reviewed. RESULTS: The two tuberculomas, histologically confirmed to have no calcification, hemorrhaging, or fibrosis, commonly showed low signal intensity on both T1- and T2-weighted images. CONCLUSION: We report two cases of a macronodular tuberculoma in the liver and kidney, which is visualized as a region of hypointensity on T2-weighted MR images. This finding appears to be important in reaching a definitive diagnosis of an abdominal tuberculoma. PMID- 8708073 TI - A segmental hyperintensity area in the liver shown by delayed MR images with Mn DPDP. AB - This is a report of a case with metastatic liver tumors in which a segmental hyperintense area was demonstrated by delayed liver MRI with manganese (II) N,N' dipyridoxylethylenediamine-N,N'-diacetate-5,5'-bis(phosphate) (Mn-DPDP). No signal abnormality except for tumors was distinct either before or 30 min after administration of Mn-DPDP. The region with hyperintensity appeared 24 h after administration. PMID- 8708074 TI - Progressive jaundice due to lymphangiosis carcinomatosa of the liver: CT appearance. AB - In a case of progressive jaundice, contrast-enhanced CT demonstrated unevenly distributed areas of periportal low attenuation throughout the liver that were demonstrated to be dilated lymphatics with multiple carcinomatous thrombi at postmortem study. This CT appearance may be characteristic of lymphangiosis of the liver. PMID- 8708075 TI - Early enhancement of tumor thrombus in the portal vein on two-phase helical CT. AB - We report a case of hepatocellular carcinoma with a portal vein thrombus that showed early arterial enhancement of dual-phase helical CT, which permitted a specific diagnosis of tumor portal venous thrombus. PMID- 8708076 TI - Spatial normalization of 3D brain images using deformable models. AB - PURPOSE: The spatial normalization and registration of tomographic images from different subjects is a major problem in several medical imaging areas, including functional image analysis, morphometrics, and computer-aided neurosurgery. The focus of this article is the development of a computerized methodology for the spatial normalization of 3D images. METHOD: We propose a technique that is based on geometric deformable models. In particular, we first describe a deformable surface algorithm that finds a mathematical representation of the outer cortical surface. Based on this representation, a procedure for obtaining a map between corresponding regions of the outer cortex in two different images is established. This map is subsequently used to derive a 3D elastic warping transformation, which brings two images into register. RESULTS: The performance of our algorithm is demonstrated on several datasets. In particular, we first test our deformable surface algorithm on MR images. We then register MR images to atlas images. In our third experiment, we apply a procedure for matching distinct cortical features identified through the curvature map of the outer cortex. Finally, we apply our technique to images from elderly individuals with substantial ventricular enlargement, and we show a good registration in the ventricular area and the surrounding brain structures. CONCLUSION: We present a highly automated methodology for spatial normalization of images, using deformable models. Applications of our methodology include stereotactic normalization of functional and structural images, morphological analysis of the brain, and computer-aided neurosurgery. PMID- 8708078 TI - Influence of spatially heterogeneous background activity on "hot object" quantitation in brain emission computed tomography. AB - PURPOSE: Our goal was to evaluate the influence of spatially heterogeneous background activity on "hot object" quantitation in brain emission CT. METHOD: We studied the effects of spatially heterogeneous background activity on hot object quantitative recovery in simulations of both spheres and realistic brain distributions (utilizing human MRI data). RESULTS: Significant underestimation of object activity concentration was seen for both cortical and subcortical hot objects, with increasing underestimation for increasing hot object/surrounding gray matter contrast. Significant "spill-in" of counts from surrounding activity was present. CONCLUSION: Hot objects are significantly influenced by both "spill out" and "spill-in." Qualitative and quantitative analyses of such objects must explicitly consider both spill-out and spill-in; this implies a correction scheme that goes beyond simple division of the observed value by a conventional recovery coefficient. PMID- 8708077 TI - Effect of geometrical distortion correction in MR on image registration accuracy. AB - In this article we investigate the effect of geometrical distortion correction in MR images on the accuracy of the registration of X-ray CT and MR head images for both a fiducial marker (extrinsic point) method and a surface-matching technique. We use CT and T2-weighted MR image volumes acquired from seven patients who underwent craniotomies in a stereotactic neurosurgical clinical trial. Each patient had four external markers attached to transcutaneous posts screwed into the outer table of the skull. The MR images are corrected for static field inhomogeneity by using an image rectification technique and corrected for scale distortion (gradient magnitude uncertainty) by using an attached stereotactic frame as an object of known shape and size. We define target registration error (TRE) as the distance between corresponding marker positions after registration and transformation. The accuracy of the fiducial marker method is determined by using each combination of three markers to estimate the transformation and the remaining marker to calculate registration error. Surface-based registration is accomplished by fitting MR contours corresponding to the CSF-dura interface to CT contours derived from the inner surface of the skull. The mean point-based TRE using three noncollinear fiducials improved 34%-from 1.15 to 0.76 mm-after correcting for both static field inhomogeneity and scale distortion. The mean surface-based TRE improved 46%-from 2.20 to 1.19 mm. Correction of geometrical distortion in MR images can significantly improve the accuracy of point-based and surface-based registration of CT and MR head images. Distortion correction can be important in clinical situations such as stereotactic and functional neurosurgery where 1 to 2 mm accuracy is required. PMID- 8708079 TI - Correction method for end-tidal xenon concentration in CBF measurements with xenon-enhanced CT. AB - PURPOSE: The goal of this work is to show how variations in respiratory rate and tidal volume affect calculated cerebral blood flow (CBF) values on xenon-enhanced CT. In xenon-enhanced CT examination, the patient often takes shallow and rapid breaths. Thus, it is less likely that end-tidal xenon concentration reflects arterial xenon concentration, and appropriate correction measures should be taken for the end-distal respiratory data to obtain reliable CBF values. METHOD: Preliminary breathing tests were performed using a lung phantom to determine the influence of respiratory volume and rate on end-tidal xenon concentration. Two xenon-enhanced CT studies were conducted of a healthy person with completely different respiratory manners between two studies. One was deep and slow respiration. The other was shallow and rapid respiration. RESULTS: The lung phantom results prove that deep and slow respiration is essential for the end tidal method. The results of xenon-enhanced CT studies of the same person show that the direct use of end-tidal data for shallow and rapid respiration leads to CBF values much lower than the actual values. CONCLUSION: Differences in respiratory rate and tidal volume during xenon inhalation can significantly affect calculated CBF values on xenon-enhanced CT. With use of the correction methods described herein, these effects can be minimized. We have derived the end tidal correction method on the assumption that a person's CBF values should be kept unchanged regardless of different respiratory manners. PMID- 8708080 TI - Aunt Minnie's corner. Nonrotation of the bowel. PMID- 8708081 TI - Purification of the bovine xanthine oxidoreductase from milk fat globule membranes and cloning of complementary deoxyribonucleic acid. AB - The amino acid sequence of the bovine xanthine oxidoreductase was determined by cloning and sequencing cDNA clones encoding the enzyme. Partial amino acid sequence corresponding to 54% of the total sequence were also determined from purified bovine milk xanthine oxidoreductase, showing identity with the translated cDNA sequence. The cDNA of 4719 nucleotides included a 5' untranslated region of 96 nucleotides, an open reading frame encoding a xanthine oxidoreductase of 1332 amino acid residues, and a 3' untranslated region of 624 nucleotides including two polyadenylation signals and a poly (A) tail of 74 nucleotides. The identity between the amino acid sequence of the bovine xanthine oxidoreductase and xanthine oxidoreductase from mammalian species was 86 to 90%. PMID- 8708082 TI - Viability of Lactobacillus acidophilus and Lactobacillus casei in fermented milk products during refrigerated storage. AB - The viability was investigated of five strains of Lactobacillus acidophilus and one strain of Lactobacillus casei that were added as adjuncts to yogurt and cultured buttermilk during 28 d of refrigerated storage at 5 to 7 degrees C. A modification of LBS (Lactobacillus selection) agar was used for the enumeration of L. acidophilus and L. casei. The medium allowed the colony formation of the adjunct bacteria while preventing colony formation of the traditional yogurt or buttermilk starter cultures. At each sampling period, colonies from the selective agar medium were isolated for confirmation of identity to confirm that only L. acidophilus and L. casei were enumerated, that their characteristics did not change during storage, or both. The strains of L. acidophilus varied in both cultured products. In buttermilk, L. acidophilus MUH-41, O-16, and L-1 exhibited no significant loss in viability, but strains 43121 and La-5 did. No significant loss in viability of L. acidophilus MUH-41 and L-1 occurred in yogurt prepared using culture CM2; however, strains 43121, O-16, and La-5 lost viability. In the yogurt prepared using culture YC-4, L. acidophilus 43121 exhibited no significant loss in viability, but MUH-41, O-16, L-1, and La-5 did. There was no loss in viability of L. casei GG during storage of any of the cultured products. PMID- 8708083 TI - Estrogen administered at final milk removal accelerates involution of bovine mammary gland. AB - To evaluate whether estrogen hastened involution of mammary tissue, Holstein cows were injected with 4 ml of ethanol excipient (n = 21) or 15 mg of estradiol-17 beta (n = 23) on each of the 4 d that preceded final milk removal. Dates of final milk removal (d 0) were designated as 60 d prior to expected dates of calving. Milk volumes were recorded, and samples were collected prior to the first and fourth injections. During the dry period, each mammary quarter within the cow was sampled once to collect secretions on dates that corresponded to d 0, 3, 11, and 25 or 1, 7, 18, and 30 of the dry period. Milk synthesis and secretion declined abruptly because of treatment. The decreased concentrations of alpha-lactalbumin, lactose, citrate, and potassium in secretions of controls, as well as the increased somatic cells, protein, lactoferrin, and sodium, occurred earlier in secretions from treated cows. These shifts of approximately 6 d, relative to days dry, suggested that exogenous estradiol increased the involution rate of mammary tissue. PMID- 8708084 TI - Effects of fenprostalene and estradiol-17 beta benzoate on parturition and retained placenta in dairy cows and heifers. AB - It was hypothesized that a high dose of estrogen in conjunction with a long acting PGF2 alpha analog would synchronize parturition within a narrow time frame and reduce the incidence of retained placenta. On d 276 of gestation, 14 animals (9 cows and 5 heifers) per group received a placebo (group A), 1 mg of fenprostalene (group B), 50 mg of estradiol-17 beta benzoate (group C), or both (group D). Treatment with estradiol-17 beta benzoate increased serum concentrations of estradiol-17 beta from 228 pg/ml at treatment to 642 and 683 pg/ml at 24 h posttreatment for groups C and D, respectively. Concentration of estradiol-17 beta in group A increased gradually to 526 pg/ml at 24 h prepartum. Progesterone concentrations were reduced by fenprostalene but not by estradiol-17 beta benzoate. Estradiol-17 beta benzoate did not reduce incidence of retained placenta in animals treated with fenprostalene (group B vs. group D) but tended to reduce incidence in uninduced animals (group A vs. group C). Thus, short-term elevation of estradiol-17 beta to normal prepartum concentrations did not regress the corpus luteum, induce parturition, or reduce incidence of retained placenta. However, elevation of estradiol-17 beta for longer periods might enhance placental separation. Conversely, fenprostalene induced calving approximately 2 d after treatment. In this study, 90% of animals treated with fenprostalene calved within a 20-h period, but with a high incidence of retained placenta. PMID- 8708085 TI - The association between cumulative milk yield, days open, and days to first breeding in New York Holstein cows. AB - The associations among milk yield, days open, and days to first breeding were studied in 15,320 Holstein cows, calving between June 1990 and November 1993, in 26 New York herds. Survival analysis, incorporating the Cox proportional hazards model, was used to quantify the effect of milk yield and other covariables on days open and days to first breeding. Sensitivity analysis was also performed. Sixty-day cumulative individual milk yield, calving season, parity, and herd were included in all models. Several diseases and their lactational risk also were studied as potential confounders: retained placenta (5%), milk fever (1%), ketosis (3%), metritis (6%), abomasal disorder (5%), mastitis (5%), and cystic ovary (3%). Cows yielding milk in the highest quintile had a slightly lower conception rate than did cows yielding milk in the lowest quintile. Conception rate varied with parity and season. Retained placenta, metritis, and cystic ovary lowered the conception rate. High yielding cows were more likely to be inseminated than were low yielding cows. Older cows and those with metritis, mastitis, and cystic ovary had lower rates on insemination. These results indicate that conception and insemination might be influenced by factors related to management (e.g., culling) and to the cow (e.g., disease history) but that increased milk yield plays a very minor role. PMID- 8708086 TI - Modeling the dynamics of radioiodine in dairy cows. AB - A metabolically based model of radioiodine transfer in cows was applied to previously collected data. The model required some modification and reparameterization before it could describe the measured data satisfactorily. In particular, significant fecal excretion of radioiodine needed to be considered. The model was used to predict the effect of dietary intake of stable iodine on the transfer of radioiodine to milk. Increased dietary intake asymptotically increased the proportion of radioiodine excreted via milk. PMID- 8708087 TI - Ruminal degradation and dose response of dairy cows to dietary L-carnitine. AB - In Experiment 1, in vitro degradation of L-carnitine was measured in ruminal fluid obtained either from a cow that was fed a 75% concentrate diet or from a cow fed a control (50% forage and 3% fat) diet. Carnitine degradation was greater in ruminal fluid from the cow fed the control diet than in ruminal fluid from the cow fed the 75% concentrate diet and was more rapid in ruminal fluid obtained after 2 wk of adaptation to dietary carnitine supplementation (7 g/d). In Experiment 2, 20 multiparous Holstein cows were used in a replicated 5 x 5 Latin square design to determine the effects of increasing the amount of dietary L carnitine (0, 0.875, 1.75, 3.5, or 7.0 g/d) that was fed to lactating dairy cows. All cows received the same diet, which contained 3% added fat. Carnitine concentration in milk and plasma increased linearly with carnitine supplementation. The DMI, milk yield, and milk composition were unaffected by carnitine. Apparent total tract digestibilities of NDF and fatty acids decreased quadratically as the amount of supplemented carnitine increased; generally, means were lowest when cows were fed 1.75 g/d of carnitine. The concentration of total VFA and molar percentages of individual VFA in ruminal fluid were unaffected by the amount of carnitine fed. Concentrations of glucose, NEFA, and urea N in plasma were unaffected by the amount of dietary carnitine; however, plasma cholesterol concentration decreased linearly as carnitine increased. Supplementation of < or = 7.0 g/d of dietary carnitine did not benefit DMI, milk yield, milk composition, or digestive measurements in this experiment. PMID- 8708088 TI - Interaction of dietary fat and acid detergent fiber diets of lactating dairy cows. AB - Twenty multiparous and 4 primiparous cows averaging 32 DIM were used in an experiment with a 2 x 2 factorial design for 12 wk to determine whether the response to supplemental choice white grease in the diet was augmented with higher fiber. Treatments were 1) 21% ADF with no added fat; 2) 21% ADF plus 6% added fat; 3) 28% ADF with no added fat; and 4) 28% ADF plus 6% added fat. Addition of fat decreased milk fat percentage and tended to decrease milk protein percentage. An interaction of fat by fiber by week for milk fat percentage suggested that ruminal fermentation was disrupted as the trial progressed when fat was included in the lower fiber diet. This disruption did not occur with the higher fiber diets. Yields of milk and FCM were not affected by treatment. There was no fat by fiber interaction for milk yield, which suggested that the yield response to fat was unaffected by fiber concentration. There were no treatment interactions for results of ruminal fermentation. Fiber increased ruminal acetate and the ratio of acetate to propionate but lowered molar percentages of propionate. Supplemental fat decreased acetate and increased propionate. The ratio of acetate to propionate was significantly decreased when fat was fed. PMID- 8708089 TI - Ruminal availabilities of protein and starch: effects on growth and ruminal and plasma metabolites of dairy calves. AB - Newborn Holstein calves (n = 75) were blocked by date of birth and sex and assigned randomly to one of eight isonitrogenous starters that contained protein and starch sources of different ruminal availabilities. Soybean meal or soybeans roasted to an exit temperature of 146 degrees C, raw or conglomerated corn, and urea at 1% of DM or no urea were used in a 2 x 2 x 2 factorial arrangement. The conglomeration process consisted of grinding the grain, adding water, pelleting the mixture, and roasting, which increased the degree of starch gelatinization fivefold. Starters were fed for ad libitum intake from 0.5 to 8 wk. Urea supplementation of conglomerated corn starters depressed performance, but the depression was greater when conglomerated corn was used with soybean meal than when it was used with roasted soybeans. Ruminal NH3 and plasma urea increased with increased RDP in starters, but the response varied according to corn type and soy protein source. Urea supplementation depressed plasma Lys, doubled plasma Cit with soybean meal and conglomerated corn starters, but depressed plasma Cit with roasted soybeans and conglomerated corn starters. Conglomerated corn depressed plasma Val and Gly, and roasted soybeans increased plasma Phe. Performance was similar when calves consumed starters containing ruminally synchronous or asynchronous CP and starch sources. PMID- 8708090 TI - Consideration of percentage of milk shipped for calculation of total lactation yields from various morning and evening plans of milk sampling. AB - Milk yield recorded on DHI test day was compared with data on milk shipped from Texas and Minnesota herds for an innovative DHI test plan referred to as alternate a.m.-p.m. without a timer. Controls were yields for test day and for milk shipped from official DHI herds in Texas, Illinois, Minnesota, and several northeastern US states. Herd milk yield for a test day as a percentage of milk shipped was considered to be an indicator of the accuracy of the DHI recording plans. Mean percentage of milk shipped was 103 for all plans and regions. When herd test days with missing values were excluded, the percentage of herd test days within 96 to 110% of milk shipped were 77 for Texas and 82 for Minnesota innovative plans and 82 for Texas, 82 for Minnesota, 79 for Illinois, and 81 for northeastern official plans. Analysis indicated that the percentage of milk shipped was consistent across herd sizes, data source, and milk yield. Eight hypothetical testing plans were examined with or without adjustment of lactation yields for percentage of milk shipped. Estimates of variance components of lactation milk yields were computed and compared using a multitrait animal model. Adjustment of records for percentage of milk shipped would decrease mean milk yields by 3%, could result in better estimates of actual milk produced, but would have little effect on accuracy of genetic evaluations. PMID- 8708091 TI - Procedures used by North Carolina dairies for vitamins A and D fortification of milk. AB - New research findings have documented fortification errors in fortified milk products all across the US milk industry. Also, the consumption of overfortified bovine milk has led the public to question whether vitamin fortification is safe. Therefore, North Carolina dairies were surveyed to determine vitamin fortification procedures used and to determine differences among these procedures. Of the parameters surveyed, the general conditions under which vitamin preparations were stored, the method used to add vitamin preparations to milk, and the point during processing at which vitamin preparations were added to milk were different among dairies. Forty-six percent of the dairies stored vitamin preparations under refrigerated conditions, and 54% stored vitamin preparations at ambient temperatures. The addition of vitamin preparations to bovine milk was accomplished by metered injection at 64% of the dairies and batch addition techniques at 36%. Vitamin preparations were added before fat content standardization and separation by 23% of the dairy processors; 77% added the vitamin preparations after this point. When dairies were asked at what point they added their vitamin preparations to the milk, nine different answers were given. Although other sources of error could also contribute to inconsistent concentrations of vitamin fortification, differences in fortification procedures may have a large impact upon the problem. The diversity of vitamin fortification practices used in North Carolina may be an indication of nationwide trends. PMID- 8708092 TI - Effects of freezing on the viability of nine pathogens from quarters with subclinical mastitis. AB - Milk samples from 45 quarters containing mastitis pathogens were collected from lactating cows to determine the viability of those pathogens after freezing. An initial bacteria count was conducted, and samples were divided into 2-ml portions and frozen. Weekly bacteria counts were conducted for 6 wk. Viability after freezing was determined on five isolates of nine bacterial species: Staphylococcus aureus, Staphylococcus hyicus, Staphylococcus chromogenes, Staphylococcus xylosus, Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus uberis, Corynebacterium bovis, and Escherichia coli. Bacteria counts were converted to logarithm base 10, and analysis of variance was conducted to determine alterations in viability over the 6-wk period. Freezing of quarter milk samples for 6 wk did not affect viability of any of these pathogens. PMID- 8708093 TI - Polymorphisms in the bovine beta-casein 5' flanking region. PMID- 8708094 TI - Purification of beta-lactoglobulin from whey protein concentrate by pepsin treatment. AB - beta-Lactoglobulin was purified from whey protein concentrate by a combination of pepsin treatment and membrane filtration. Porcine pepsin was added to whey protein (1:200, wt/wt), and the mixture was then incubated at pH 2.0 and 37 degrees C for 60 min. The protein fraction was collected by ammonium sulfate precipitation, and the precipitate was either dialyzed against water using a dialysis membrane (20-kDa pore size) or filtered using an UF membrane (30-kDa pore size). The beta-LG did not differ from standard beta-LG as measured by chromatography, SDS-PAGE, native PAGE, differential scanning calorimetry, or UV spectrum. Based on the results, a simplified procedure was developed, consisting of pepsin treatment and UF, to purify beta-LG directly from whey. PMID- 8708095 TI - Purification and characterization of a dipeptidase from Pseudomonas fluorescens ATCC 948. AB - A dipeptidase from a cell extract of Pseudomonas fluorescens ATCC 948 was purified to homogeneity by ion-exchange chromatography, hydroxyapatite chromatography, gel filtration, and FPLC Phenyl Superose chromatography. The enzyme was located in the cytoplasmic fraction. The dipeptidase appeared to be a monomer with a molecular mass of about 46 kDa, and activity was optimal at pH 8.0 and 50 degrees C. Although activity decreased markedly at temperatures > 50 degrees C, about 50% of maximal activity was retained at 10 degrees C. The activity was inhibited by EDTA, but could be reactivated with Co2+ and partially reactivated with Ca2+. Reducing agents, such as dithiothreitol and 8 hydroxyquinoline, also strongly inhibited the enzyme. The dipeptidase hydrolyzed only dipeptides and showed a particular aptitude for substrates containing a hydrophobic AA at the N-terminus. Dipeptides containing a proline residue were not cleaved. Kinetic studies indicated that the dipeptidase hydrolyzed Leu-Leu with a Michaelis-Menten constant of 0.43 mM and a turnover number of 1812/s. PMID- 8708096 TI - Glucose and xylitol tolerance tests for ketotic and healthy dairy cows. AB - Intravenous administration of 500 ml of 50% glucose solution to 10 nonketotic dairy cows increased the blood glucose and insulin concentrations 7-fold immediately following administration. Blood glucose and insulin concentrations of ketotic cows were about 6- and 3-fold higher, respectively, immediately following glucose administration. Administration of 1000 ml of 25% xylitol (xylo-pentane 1,2,3,4,5-pentol) in nonketotic cows increased blood glucose and insulin concentrations 2- and 9-fold, respectively. Ketotic cows treated with xylitol exhibited blood insulin concentration 12-fold higher following administration. This insulin increase might be explained by a decrease in insulin degradation because of the diffusion of xylitol, which is not dependent on insulin in peripheral tissues. For ketotic cows given xylitol, serum concentrations of free fatty acid decreased, and triglyceride concentrations and aspartic acid aminotransferase activity increased, but values were unchanged by xylitol administration to nonketotic cows. Thus, for ketotic cows, the responses of the blood glucose and insulin concentrations to xylitol administration were better than those responses to glucose administration. Improvements in clinical signs, i.e., disappearance of urinary ketone bodies and recovery to normal feed consumption, also suggested the usefulness of xylitol administration for the treatment of ketosis. PMID- 8708097 TI - Field trials of an oral calcium propionate paste as an aid to prevent milk fever in periparturient dairy cows. AB - Trials were conducted to test the efficacy of a calcium propionate paste as an aid to prevent milk fever and to improve the health of dairy cows. Each calcium propionate treatment tube supplied 37 g of calcium. In trials involving Holstein herds and a Jersey herd, two (trials 1 and 2) or three (trial 3) calcium propionate tubes were given at calving and again at 12 h after calving. For the Jersey herd, calcium propionate treatment (two tubes) reduced the incidence of milk fever from 50% in control cows to 29% in treated cows. Plasma obtained 24 h after calving from treated cows had higher calcium, lower NEFA, and lower beta hydroxybutyrate concentrations than did plasma from control cows. No other benefits of calcium propionate treatment were significant for health or for productivity of the cows. Calcium propionate treatment had no significant effects on blood calcium, NEFA, or beta-hydroxybutyrate in the Holstein herds studied. However, calcium propionate did reduce the number of cows with subclinical hypocalcemia (< or = 7.5 mg/dl of plasma calcium) at 24 h after calving in both trials involving Holstein cows. Calcium propionate treatment was beneficial in reducing subclinical hypocalcemia in all trials and reduced the incidence of milk fever in a herd having a problem with milk fever. PMID- 8708098 TI - Methodology of comparing three milking clusters in a large dairy herd. AB - Data for milking parameters were obtained from three different milking clusters using a modified Latin square design to take into account the real milking time regimen of the farm and the time requirements to sort cows. In this modification, cows entered the milking parlor in a random sequence rather than by group. In addition, the clusters were assigned to milking sites. Parameters were compared for 554 cows from one dairy herd that was divided into two smaller herds of 410 and 144 cows. A modified Latin square design was used to compare clusters. The results revealed that 92.1 and 95.5% of cows in Experiments 1 and 2, respectively, were treated by all three clusters; 7.9 and 4.5% of the cows, respectively, were treated by only two clusters; and no cow was treated by only one cluster. The milking parameters for both experiments were in good agreement. Results demonstrated that the modified Latin square design was equivalent to the original Latin square design under the constraint that the number of cows included in one experiment should be > 100 and that the number of milkings should not be < 200 per cluster. PMID- 8708099 TI - Measurement of liner slips, milking time, and milk yield. AB - Liner slip or rapid air leakage past the mouthpiece of the milking machine liner is related to high rates of new cases of mastitis. A real time technique was developed to monitor the air flow into the milking machine cluster during liner slips as well as to monitor milking time and milk yield using a commercial type pipeline milking system. The air flow into the cluster was measured by recording the pressure differences across an orifice plate placed in the air bypass of an air-milk separator using a differential pressure transducer. Milk yield was recorded by counting the number of milk releases from an electronic milk meter. The release solenoids of the milk meter were linked to a computer. The start and end of milking were manually recorded by switching a two-pole switch connected to a digital input card on the computer, which was programmed to record air flow, milk yield, and milking time. Milk yield, milking time, and air flows during liner slips were recorded simultaneously at each milking unit in an 11-unit herringbone parlor. The system was tested with an experiment with a 4 x 4 Latin square design using four treatments (clusters) and four treatment groups (22 cows per group). PMID- 8708100 TI - Economic benefits of reducing somatic cell count under the milk quality program of Ontario. AB - An empirical stochastic model for monthly bulk milk SCC was used to determine the impact of bulk milk SSC on the probability and cost of penalties under the milk quality program of Ontario, Canada. Under this program, milk price is reduced by $1 (Canadian)/hl each time bulk milk SCC exceeds 500,000 cells/ml three times in a 4-mo period. The model included herd average bulk milk SCC, effects of season, five autoregressive terms for bulk milk SCC in previous months, and a random variable with a standard deviation that depended on herd average SCC. Probabilities of receiving a penalty were 20, 13, 7, 4, 1.5, and 0.4%/mo for herd average bulk milk SCC of 450,000, 400,000, 350,000, 300,000, 250,000, and 200,000 cells/ml, respectively. Associated average penalty costs were $0.81, 0.47, 0.17, 0.04, 0.002, and 0.000/hl, respectively. Marginal return functions for reducing herd average linear somatic cell score were derived also. Marginal values were minimal for herd average linear scores < 3. For the population, the marginal value of reducing average linear score by one unit from a mean of 3 was $19.60/yr per cow. Results indicated that efforts to reduce bulk milk SCC resulted in substantial extra milk revenues. A bulk milk SCC target of 250,000 cells/ml was advocated. PMID- 8708101 TI - Altering conception of dairy cattle by gonadotropin-releasing hormone preceding luteolysis induced by prostaglandin F2 alpha. AB - The objective of these experiments was to determine the effect on fertility of GnRH when used in conjunction with one or two injections of PGF2alpha. In Experiment 1, GnRH was administered 7 d before the second of two injections of PGF2alpha (14 d apart). The control group received two injections of PGF2alpha without GnRH. Conception was reduced from 63.5% for 74 controls to 48.7% for the 79 heifers and cows that had been treated with GnRH, but estrus detection and pregnancy rates were similar. In Experiment 2, 85 heifers and cows received GnRH at a random stage of the estrous cycle, followed in 7 d by PGF2alpha. Thirty to 32 h after PGF2alpha, a second dose of GnRH was given to induce ovulation of the preovulatory follicle, followed by one fixed-time insemination 18 to 19 h later (treatment designated as GnRH, PGF2alpha, and GnRH). Controls (n = 85) were given PGF2alpha and inseminated at estrus. Although conception rate was not different, one fixed-time insemination after the GnRH, PGF2alpha, and GnRH treatment tended (35.3%) to reduce fertility compared with effects of the control (47.1%). It is unclear how an injection of GnRH during the intervening week between two injections of PGF2alpha reduced fertility in Experiment 1. However, in Experiment 2, when GnRH was given 7 d before one injection of PGF2alpha and when ovulation was induced with a second GnRH injection, one fixed-time insemination seemed to produce acceptable fertility in dairy cows but probably less than that when inseminations were based on detected estrus. PMID- 8708102 TI - Effects of Saccharomyces cerevisiae and Aspergillus oryzae cultures on ruminal fermentation in dairy cows. AB - Four lactating Holstein cows, fitted with ruminal and duodenal cannulas, were used in a 4 x 4 Latin square design to examine the effects of supplemental yeast (Saccharomyces cerevisiae) and fungal (Aspergillus oryzae) cultures on ruminal fermentation, microbial populations, and nutrient supply to the small intestine. Cows were fed a basal diet comprising 32.5% corn silage, 17.5% alfalfa hay, 35.3% corn grain, 12.7% soybean meal, and 2% vitamin and mineral mixture on a DM basis. Treatments were arranged in a 2 x 2 factorial as follows: 1) basal diet, 2) basal diet plus 57 g/d of yeast culture, 3) basal diet plus 3 g/d of fungal culture, and 4) basal diet plus 57 g/d of yeast culture and 3 g/d of fungal culture. Ruminal pH, ammonia N concentration, and total VFA concentration were similar among treatments. Molar percentages of ruminal isoacids were lower for cows fed a mixture of yeast and fungal culture than for cows fed yeast or fungal culture alone. Yeast culture increased ruminal OM and CP digestion and decreased OM and N flow to the duodenum. Fiber digestion in the rumen was similar among treatments. Fungal culture stimulated proteolytic and cellulolytic bacterial counts. Proteolytic bacterial counts were also stimulated by yeast culture. Results from this experiment demonstrated that yeast and fungal cultures could influence ruminal fermentation and microbial populations. PMID- 8708103 TI - Enhanced intake and production of cows offered ensiled alfalfa with higher neutral detergent fiber digestibility. AB - The objective of this study was to determine the effect of forage fiber digestibility as part of a total mixed diet on intake and production of cows with intakes that were likely limited by rumen fill, using treatments that were not confounded by fiber source, concentrate, or ratio of forage to concentrate. Two alfalfa silages with similar NDF concentrations (40%) but different NDF digestibilities (40 vs. 45% after 24 h of in vitro fermentation) were harvested, mixed with concentrate to achieve diets containing 35% NDF, and offered to 12 multiparous cows (13 DIM) in a two-period balanced crossover design. Samples taken during silage feeding revealed that diets differed in both NDF digestibility (3 percentage units) and NDF content (1.8 percentage units), making interpretation of results difficult. Nonetheless, milk production (36.3 vs. 38.2 kg/d) and DMI (19.4 vs. 20.4 kg/d) were significantly higher with the higher NDF digestibility diet. Apparent in vivo DM and NDF digestibilities, total rumen VFA after feeding, and molar percentage of propionate were also higher with this diet. Equal NDF intake between diets suggested that higher DMI was due to the lower NDF content in the more digestible NDF diet. Higher NDF digestibility might also increase DMI; further experimentation is necessary with larger unconfounded differences in fiber digestibility at the time of feeding. PMID- 8708104 TI - Fatty acid digestibility and lactation performance by dairy cows fed fats varying in degree of saturation. AB - Holstein cows fed fats varying in degree of saturation were used to evaluate lactation performance (35 multiparous and 15 primiparous cows) and fatty acid digestibility (5 cows). Data from wk 2 and 3 of lactation were used for covariable adjustment of data from wk 4 through 19. Diets were a basal diet (control) with no added fat and four diets with 5% added fat from tallow, tallow plus partially hydrogenated tallow in proportions of 2:1 or 1:2, or partially hydrogenated tallow; iodine values were 45, 35, 26, and 16 for the diets with added fat, respectively. Digestibilities of OM, NDF, and N were not affected by fat supplementation (mean of four fat treatments vs. control) or by degree of fat saturation. Fatty acid digestibility was lower for cows fed fat than for those fed the control diet and decreased linearly with increased fat saturation. In both trials, DMI increased linearly as fat saturation increased. In the digestion trial, cows fed fat tended to have lower DMI than those fed the control diet, primarily because of the unsaturated fat. In the production trial, DMI was similar for cows fed the control diet (22.3 kg/d) or the diet with added fat from tallow (22.1 kg/d) and tended to be higher for cows fed the diet containing partially hydrogenated tallow (23.9 kg/d). Milk production was higher for cows fed fat than for cows fed the control diet, but 4% FCM was unaffected. Milk fat and protein percentages were not affected by fat supplementation but increased linearly with increased fat saturation. Mean body condition score of cows increased as fat saturation increased. The lower digestibility of the diet with added fat from partially hydrogenated tallow was offset by higher DMI and percentages of milk fat and protein as fat became more saturated. PMID- 8708105 TI - Occurrence of conjugated cis-9,trans-11-octadecadienoic acid in bovine milk: effects of feed and dietary regimen. AB - Twenty-eight lactating cows were used to study the effects of different diets and different dietary regimens on the variation of cis-9,trans-11-octadecadienoic acid in milk. Cows were divided into three groups and assigned to dietary treatments differing in the ratio of forage to concentrate. The control group was fed a diet with an average ratio of forage to concentrate of 50:50. The two trial groups were fed a trial diet either in restricted or nonrestricted amounts. Variation of cis-9,trans-11-octadecadienoic acid in milk was substantial (2.5 to 17.7 mg/g of fat). Cows fed restricted amounts of the trial diet, which contained a lower forage to concentrate ratio than did the control diet, exhibited, on average, the highest concentration (11.28 mg/g of fat). For cows fed for ad libitum consumption, the trial diet did not differ significantly from the control diet (6.6 vs. 5.0 mg/g of fat). The concentration of cis-9,trans-11 octadecadienoic acid and trans-11-C18:1 was positively correlated. Therefore, conjugated linoleic acid in bovine milk, if considered to be beneficial, could be increased through a suitable dietary regimen. PMID- 8708106 TI - Response of early lactation cows to ruminally undegradable protein in the diet. AB - The objective of this study was to determine the response of dairy cows in early lactation to diets based on the recommendations of the Agricultural Research Council or the NRC for sustaining milk yield. Diets were formulated to satisfy the nutrient requirements of Holstein cows weighing 600 kg and yielding 35 kg of 3.5% fat milk/d according to either Agricultural Research Council recommendations or NRC recommendations for RUP. A third diet was a 1:1 (wt/wt) mixture of the Agricultural Research Council and the NRC diets. The same forage was fed in all diets at a forage to concentrate ratio of 40:60, and the RUP supply was altered by substituting fish and corn gluten meal for canola meal. The calculated effective degradabilities of CP for the TMR were 70.1, 66.1, and 62.1% for the Agricultural Research Council diet, the mixture of Agricultural Research Council and NRC diets, and the NRC diet, respectively. Milk composition was similar for the three dietary treatments. Multiparous cows showed a linear yield response (30.4, 31.6, and 33.7 kg/d) to increasing inclusion of the NRC concentrate in the diet. No response to additional RUP was observed for first lactation heifers. Agricultural Research Council recommendations for dietary RUP underestimated the requirements for multiparous cows in early lactation yielding > 25 kg of milk/d. PMID- 8708107 TI - Apparent digestibility of minerals by lactating cows from a total mixed ration supplemented with poultry litter. AB - Apparent digestibility of minerals by lactating cows from a multiple-component, conventional Israeli TMR (control) and from a TMR containing 10% poultry litter (DM basis) was examined. Ten cows in early lactation were divided into two groups of similar performance (39 kg of milk/d) and fed for ad libitum intake the two TMR for 28 d. Poultry litter contributed, as a percentage of total requirements, 44.4% of Ca, 41.0% of P, 32.0% of S, 22.8% of Mg, and 25.9% of K. Apparent digestibility of the macroelements was higher for cows fed the TMR supplemented with poultry litter than for those fed the control TMR. Apparent digestibility of the control TMR was 22.3% for Ca, 29.4% for Mg, 39.1% for P, and 62.2% for S; the apparent digestibility of the treatment TMR was 30.1% for Ca, 45.6% for Mg, 45.3% for P, and 65.6% for S. Poultry litter contributed the entire requirements of Zn, Mn, and Co; 56% of Cu; and 32% of Se. The apparent digestibility of the control TMR was 15.6% for Cu, 39.6% for Zn, 8.51% for Mn, 42.8% for Co, and 41.6% for Se, and the apparent digestibility of the treatment TMR was 27.9% for Cu, 54.0% for Zn, 17.8% for Mn, 37.0% for Co, and 63.9% for Se. Poultry litter is a good source of macro- and microelements for lactating cows and, at 10% of the TMR, could ensure against mineral deficiencies. PMID- 8708108 TI - Effect of supplemental lactoferrin with ferrous iron on iron status of newborn calves. AB - Data for 18 male and 18 female calves, born from primiparous and multiparous cows, were collected to determine the effect of supplemental lactoferrin and FeSO4 for 5 d after parturition on the Fe status of calves. Dietary treatments were 1) untreated, 2) 40 mg of Fe/d as FeSO4, and 3) 40 mg of Fe as FeSO4 plus 5 g of lactoferrin/d. Blood hematocrit and hemoglobin of calves born from primiparous cows at d 1 of age were lower than those of calves born from multiparous cows, but not different from those of their dams. Blood hematocrit and hemoglobin of male calves at d 1 of age were lower than those of female calves. Plasma Fe of primiparous cows at parturition was lower than that of multiparous cows, but plasma Fe of calves was not affected by parity of dam and sex of calf. Blood hematocrit and hemoglobin of untreated calves decreased from 1 to 10 d of age. Blood hematocrit and hemoglobin of calves treated with Fe of Fe plus lactoferrin increased from 2 to 10 d of age. Blood hematocrit and hemoglobin of calves treated with Fe plus lactoferrin were higher than those of calves treated with Fe at d 6 of age. Plasma Fe of calves treated with Fe or Fe plus lactoferrin increased temporarily at d 2 of age. Plasma Fe of calves treated with Fe plus lactoferrin at d 2 of age was lower than that of calves treated with Fe, but, at d 6 and 10 of age, plasma Fe of calves treated with Fe plus lactoferrin were higher. PMID- 8708109 TI - Effect of roasted soybeans and corn on performance and ruminal and blood metabolites of dairy calves. AB - Newborn Holstein calves (n = 132) were used to evaluate three protein sources (soybean meal and soybeans roasted to an exit temperature of 138 or 146 degrees C) that contained 30, 45, or 52% RUP, respectively. Each was used with either raw corn or corn roasted to an exit temperature of 135 degrees C. Roasting of corn increased starch gelatinization from 34.8 to 118.6 mg of maltose equivalents/g of sample. Six pelleted isonitrogenous calf starters (18% CP) were fed for ad libitum consumption from 0.5 to 8 wk of age. Performance of calves fed soybeans roasted at 146 degrees C plus raw corn was superior to that of calves fed soybean meal plus raw corn but was similar to the performance of calves fed soybean meal plus roasted corn. Roasted corn tended to improve calf performance when used with soybean meal and, to a lesser degree, when used with soybeans roasted at 138 degrees C but depressed performance when used with soybeans roasted at 146 degrees C. Ruminal concentrations of NH3, butyrate, and total VFA and plasma concentrations of urea were higher for calves fed soybean meal. Ruminal pH and lactate, plasma NEFA, and blood BHBA concentrations were unaffected by starter. Performance was superior when calves consumed protein and corn sources with similar degrees of ruminal availability. PMID- 8708110 TI - Analytical criteria for predicting apparent digestibility of soybean protein in preruminant calves. AB - A series of experiments on the use of soybean as a protein source in milk replacers for veal calves was undertaken to determine the relationships between the physicochemical and antinutritional properties and apparent digestibilities of nine soybean products. Soybean provided between 58 and 71% of dietary CP, and skim milk or whey powder provided the remainder. Soybean products were analyzed for CP, native protein, aggregated protein, carbohydrate-linked protein, peptides, antitryptic activity, and immunoreactivity of lectin, glycinin, alpha conglycinin, and beta-conglycinin. The apparent digestibility of N calculated for soybean protein varied between 59 and 84%. Simple linear correlations were significant between apparent digestibility of soybean N and concentrations of native protein, antitryptic activity, glycinin, alpha-conglycinin, and beta conglycinin. However, only variation in antitryptic activity, alpha-conglycinin, and beta-conglycinin contributed significantly to an explanation for the variation in apparent digestibility of soybean N in a multiple linear equation. Under our experimental conditions, beta-conglycinin was the best predictor of digestibility of soybean N. Antitryptic activity became the best predictor when soybean products had no detectable beta-conglycinin. PMID- 8708111 TI - Effect of Lactobacillus acidophilus supplementation of milk replacer on preweaning performance of calves. AB - Forty Holstein-Friesian calves were used to evaluate the effect on young calves of daily dietary supplementation with Lactobacillus acidophilus. Calves were randomly assigned at 2 d of age to one of two treatments; 1) milk replacer with no additives or 2) milk replacer supplemented with 1 ml (5 x 10(7)) of viable L. acidophilus bacteria at each of two feedings per day. Milk replacer was reconstituted to 12% DM and fed at 10% of BW/d for the duration of the 6-wk trial. A commercial starter pellet was offered for ad libitum intake from 7 d of age. Treatment had no effect on actual BW at any stage or on total BW gain; however, average daily gain during wk 2 was affected by L. acidophilus supplementation. Calves receiving L. acidophilus maintained initial BW, and the control calves lost BW until 2 wk of age, at an average rate of 112 g/d. Starter intake, total DMI, feed efficiency, and occurrence of diarrhea were unaffected by treatment. Therefore, L. acidophilus supplementation for calves fed milk replacer may be beneficial during the first 2 wk of life. PMID- 8708112 TI - Estimates of genetic trend for yield traits of the registered Jersey population. AB - Data for American Jersey cattle from 1960 to 1990 were used to study genetic progress achieved by four paths of selection (sires of bulls, dams of bulls, sires of cows, and dams of cows). Possible methods of obtaining optimal genetic gain for the breed were suggested by the analysis. The generation interval by year of birth for parents of AI bulls was greater from 1975 to 1984 than for most recent 5 yr (1983 to 1987) because previously the use of old proven bulls was greater. Sires of bulls tended to have a longer average generation interval (9.3 yr) than the more nearly optimal interval reported in the literature for AI populations (7.0 yr). Selection differentials for the last 5 yr were larger than the average over all years for milk yield, fat yield, protein yield, and fat percentage. More intense selection for sires of bulls gave a larger increase in PTA over time than did other paths of selection. When all paths of selection were combined, estimated annual genetic change for the Jersey breed was greater from 1983 to 1987 than from 1960 to 1987 for milk yield, fat yield, protein yield, and fat percentage. Potential genetic response from the sires and dams that were available to produce sons was greater than the response from parents actually used. Therefore, better sires and dams were available for genetic enhancement of their progeny than those actually used to produce young sires and replacement females. PMID- 8708113 TI - Genetic analysis of growth patterns of black and white dairy heifers. AB - Monthly BW to 600 d of age along with BW at first calving of 743 heifers were analyzed to estimate the genetic relationship between BW at first calving and other growth variables. The Von Bertalanffy function and a two-phase logistic function were used to fit growth patterns of individual heifers. The two-phase logistic function fitted the data more adequately than did the Von Bertalanffy function. Calves from first parity dams were born at a lower degree of maturity than calves from third parity dams. Calves with a high proportion of Holstein genes had a higher BW both at first calving and at maturity. Heritabilities of the variables of the Von Bertalanffy and the two-phase logistic function ranged from 0.02 to 0.31 and 0.15 to 0.26, respectively. Estimated mature BW had a highly negative correlation with rate of maturation (-0.56 to -0.82); BW at first calving had a high genetic correlation (0.74 to 0.90) with estimated mature BW and a negative correlation with maturation rate (-0.18 to -0.40). Results of this study indicated that BW at first calving can be used as an indirect trait when mature BW is included in the breeding goal. PMID- 8708114 TI - Estimates of inbreeding and relationship among registered Holstein females in the United States. AB - The estimate of inbreeding for registered Holstein females born during 1990 was 5.1 +/- 0.4% for a random sample of 600 two-line pedigrees. Previously published estimates were 4.7% for 1970, 3.8% for 1976, and 4.3% for 1982 from similar samples of pedigrees. These estimates were calculated using a base of approximately 1884, or the initiation of herd-book registration in the US. The estimate of relationship between randomly paired females born during 1990 was 10.2 +/- 0.8%; relationships were 5.2% for 1970, 7.3% for 1976, and 9.8% for 1982. Average relationship within the breed increased significantly without a corresponding increase in inbreeding. Inbreeding was also estimated for alternative base years with 10-yr increments to determine the effect of base year on estimates of inbreeding. Using 1950 as the base year, inbreeding was estimated at 0.5% for 1970, 0.3% for 1976, 1.6% for 1982, and 2.1% for 1990. Estimates using 1920 as the base year were 2.0% for 1970, 1.9% for 1976, 2.3% for 1982, and 3.2% for 1990. More recent base years gave lower estimates of inbreeding, as expected, but showed an exaggerated percentage of increase in inbreeding from 1970 to 1990. However, the annual amount of increase of inbreeding from 1976 to 1990 was not significantly affected by time of base year and ranged from 0.08 to 0.12/yr. The five most influential bulls of the breed were Pawnee Farm Arlinda Chief 1427381, Round Oak Rag Apple Elevation 1491007, S-W-D Valiant 1650414, Osborndale Ivanhoe 1189870, and Sir Pietertje Ormsby Mercedes 44931; estimated direct relationships to the breed were 0.123, 0.122, 0.096, 0.083, and 0.074, respectively. PMID- 8708115 TI - Dairy operation management practices and herd milk production. AB - A national US survey collected data on herd milk production and management of Holstein herds. Step-wise selection identified management practices that were related to herd milk production using only operations that calculated herd milk production as well as using data from all operations. Results were similar. Milk production was highest in the West. Operations with < 100 dairy cows had lower production than did larger operations. Operations with > 25% registered cattle had higher production than operations with no registered cattle. Dairy operations that reported a mean BW > 545 kg at first calving had higher mean milk production than operations with a mean BW < or = 545 kg at first calving. Operations that reported mean age at first calving < 27 mo had higher mean milk production than operations with a mean age > or = 27 mo at first calving. In addition, use of the following management practices was associated with higher rolling herd average milk production: calves born in individual areas in buildings, calves hand-fed first colostrum, starter grain fed to preweaned calves, ionophores fed to heifers from birth to first calving, DHIA record-keeping system used, computerized records, and no new cattle introduced in the previous 12 mo. PMID- 8708116 TI - A conservative method for the treatment of undermining resorption of a mandibular primary molar. AB - The authors describe a simple, conservative procedure to guide the first permanent molar into its normal position in the dental arch. The second primary molar is recontoured and covered with a stainless steel crown, thus providing a free path of eruption for the first permanent molar. PMID- 8708117 TI - That acquired masseteric look. AB - There is a certain "look" that has become the ideal of attractiveness in our society. This appearance may be due to masseteric hypertrophy, which can have an extraordinary effect on facial form. A case is presented and discussed. The practitioner should be alert to investigate further when treating patients with this look. Thorough medical and dental histories are essential in establishing this diagnosis. PMID- 8708118 TI - Prevalence and treatment costs of infant caries in Northern California. AB - To determine the prevalence of infant caries in low-socioeconomic-status children and to analyze the costs of their treatment, we retrospectively evaluated 357 children, ages eight months to seven years, who were treated during 1992 at a university-associated medical center in Northern California. Infant caries was diagnosed by several different standards, and prevalence varied according to the diagnostic criteria employed: 27 percent by the presence of any labiolingual lesion on the maxillary incisors; 32 percent by the presence of at least two carious maxillary incisors; 27 percent by the presence of at least three carious maxillary incisors; and 36 percent by a dmft > or = 5. Prevalence was higher among boys than girls (37 percent versus 27 percent), and highest in the group ages 3-4 years (43 percent). The cost of dental treatment increased with deft and ranged from $408 for deft 2-5 to $1725 for deft 16-20. Many patients failed to comply with recommended treatment for reasons of cost. Our results showed that (1) the prevalence of infant caries varies depending upon the clinical criteria used for diagnosis; (2) the cost of rehabilitating primary dentition increases in proportion to the number of teeth involved; and (3) low-income patients avoid treatment of infant caries for a variety of reasons related to the cost involved. PMID- 8708119 TI - A prospective study of the feeding and brushing practices of WIC mothers: six- and twelve-month data and ethnicity and familial variables. AB - This study, part of a project that provided a baby cup to WIC children in Washington State, reports the feeding and brushing behaviors at six and twelve months and the influence of cultural and familial variables. Results show there are large differences in feeding and brushing practices among ethnic groups, i.e., Asian mothers reported the greatest reliance on the bottle at twelve months. Marital status was related to feeding practices, i.e., single women were more likely to put the baby to bed with a bottle at six and twelve months and were more likely to use a cup at twelve months. The presence of additional children at home was related to infrequent brushing and lower rates of cup use at twelve months. Awareness of ethnic and familial variables such as marital status and number of children may be useful in formulating specific recommendations when counseling mothers and other caretakers. PMID- 8708120 TI - Changes in urinary excretion of catecholamines and their metabolites in pediatric dental patients. AB - To explain the changes in urinary excretion of catecholamines and their metabolites in pediatric dental patients, comparative measurements of three catecholamines (dopamine, noradrenaline and adrenaline), and five metabolites (3, 4-dihydroxyphenylacetic acid, normetanephrine, metanephrine, homovanillic acid, and vanillylmandelic acid) were made before and after dental treatment. The investigations were carried out in seven healthy young children between three and five years of age. All these children had previously displayed negative behavior in dental treatment. There were statistically significant increases (P < 0.05) in urinary noradrenaline, adrenaline, and normetanephrine levels during dental treatment. These results suggest that these three substances could be a useful tool for determining the occurrence of dental stress in children. PMID- 8708121 TI - Retention of a maxillary primary canine: fifty years above and beyond the call of duty. AB - A case is reported of a retained maxillary primary canine that functioned effectively over fifty years. At the time of exfoliation the tooth was still of normal length and was lost not because of ankylosis and root resorption, but because of periodontal disease. Excellent histological preservation of the specimen at the time of exfoliation, together with a very complete clinical history, permitted the development of a detailed description of a tooth and its dying pulp while under function over a long period of time. PMID- 8708122 TI - Oral appliance attached to a bubble helmet for prevention of self-inflicted injury. AB - A six-year-old girl was referred to the Dental Clinic of Monroe Developmental Center by the New York State School for the Blind. She had a history of self abusive behavior, particularly in biting her lower lip, and had previously had all primary teeth removed to prevent further damage. The school psychologist was concerned about new injuries because her permanent first molars were erupting and cheek lacerations were already present in those areas. An oral appliance attached to a bubble helmet was designed and fabricated. The patient tolerated it well. Her behavior improved with a change in psychotropic medication, and the lacerations of oral and peri-oral areas healed. At this time, more permanent teeth are present in the oral cavity. PMID- 8708124 TI - Dental status and fluoride consumption among five-year-old schoolchildren in Strasbourg, France. AB - An epidemiological survey of the caries prevalence in 1743 five-year-old children was conducted in Strasbourg in 1991 and compared with a similar study performed in 1973 (Jung et al, 1975). In 1991, 44.2 percent of the children were caries free and 3 percent of the sample had experienced extractions of carious primary molars. The average values of the dfs and dft indices were 2.93 and 6.10, respectively, with a caries reduction of 35 percent when compared to the 1973 data. The majority of children (87.96 percent) used fluoridated dentifrices and 49.5 percent of them benefited from at least another combined fluoride supplementation. The latter showed lower caries indices than the unsupplemented ones. The average plaque index and gingival index values were 0.62 and 0.1, respectively: 46.4 percent of the children had a plaque index of 0. PMID- 8708123 TI - Managing a peripheral ossifying fibroma. AB - The Peripheral Ossifying Fibroma is an inflammatory lesion which most often appears in twenty-five to thirty-four-year-old females. It averages 1.0 cm at its greatest dimension. This case reports a seven-year-eight-month-old female who presented with a peripheral ossifying fibroma lesion which measured 2.7 cm by 1.5 cm by 1.0 cm. A review of peripheral ossifying fibroma, and the management and postsurgical sequelae of this child are discussed. PMID- 8708125 TI - Aid to families with dependent children: who receives more than $22 billion and why? AB - A general outline of the Aid to Families with Dependent Children program is provided. The $22 billion program provides financial support to 14 million persons (including more than 9 million children). The changing character of the family structure is considered in terms of efforts to control AFDC spending. Additional programs to assist children (Social Security, Supplemental Security Insurance and Food Stamps) are reviewed. PMID- 8708126 TI - Pediatric AIDS epidemic reflects the first half million AIDS cases in the U.S. AB - Almost seven thousand pediatric AIDS cases with a death rate of 55 percent have been reported. The continuing increase and spread of this epidemic to different child populations and areas in the country is reviewed. Because the progress of this epidemic, pediatric practitioners require an awareness of the evolving epidemiologic patterns require an awareness of the evolving epidemiologic patterns in both the pediatric and adult populations. PMID- 8708127 TI - A randomized double-blind trial of chloral hydrate/hydroxyzine versus midazolam/acetaminophen in the sedation of pediatric dental outpatients. AB - The purpose of this study was to evaluate in a randomized, double-blinded fashion the sedative and behavioral responses of children undergoing pediatric dental procedures as outpatients and who had received a combination of either oral chloral hydrate 50 mg/kg, not to exceed 1 gm, with hydroxyzine 25 mg (Group 1); or oral midazolam 0.5 mg/kg with acetaminophen elixir 10 mg/kg (Group 2). The children were rated according to sleep, movement, crying and overall behavior at preselected intervals that included presedation, introduction to the operating suite, injection, and overall behavior. There were no statistical differences between the two groups in regards to age, weight, length of procedure, or sex. Group 1 was more asleep in the later time intervals (P < 0.05). Their were no statistically significant differences between the groups in relation to the overall evaluation for the procedure. In conclusion, midazolam/acetaminophen is as effective as chloral hydrate/hydroxyzine in the sedation of young children undergoing outpatient dental procedures. PMID- 8708128 TI - Effects of Actinomyces amphiphile on the fluidity of endothelial cells: a spin label study. AB - Actinomyces amphiphile (AcA) is an amphipathic molecule produced by Actinomyces viscosus that exhibits several biological activities. The effect of AcA on the fluidity and permeability of the plasma membrane in human umbilical vein endothelial cells was analyzed by a spin label method with 5- and 16-stearic acid nitroxide labels (SAL). These labels help to visualize the fluidity at the shallow (5-SAL) and deep (16-SAL) portions of the lipid bilayer. Cells were incubated with and without AcA (control) at 37 degrees C for 6 hours, and membrane fluidity was periodically measured. Another spin label, 4-(N, N-dimethyl N-hexadecyl) ammonium-2, 2, 6, 6-tetramethyl-piperidine-1-oxyliodine (CAT-16), was also used to assess the physical state of the cell surface. The order parameter of 5-SAL was significantly lower in the cells incubated with AcA than in control cells after the six-hour incubation. The motion parameter of 16-SAL was significantly lower in AcA-treated cells than in controls after 4 and 6 hours of incubation. These findings indicated that the AcA increased the fluidity. There were no significant differences between the AcA-treated and control cells incubated for only 2 hours. In addition, there were no differences in CAT-16 measurements between AcA-treated and control cells. The release of endoplasmic lactate dehydrogenase (LDH) into the medium tended to increase in the AcA-treated vs. the control cells. LDH release increased in both a dose- and time-dependent manner, indicating that AcA increased the permeability of plasma membranes. These findings suggest that AcA alters the biophysical properties of the plasma membranes of endothelial cells, affecting membrane function. PMID- 8708129 TI - The final pH of bacteria comprising the predominant flora on sound and carious human root and enamel surfaces. AB - Acidogenesis at low pH appears to be an important bacterial cariogenic trait. However, most information in this regard pertains to only a few of the acidogenic dental plaque bacteria. Therefore, the 'final' pH in sugar broth was determined for a wide variety of oral bacteria. Their source was: (1) carious material from advanced root lesions (ARL), (2) plaque from sound root surfaces of root-caries free subjects (SRS), (3) plaque from "white spot" coronal lesions and sound coronal surfaces of caries-active subjects, and (4) plaque from sound coronal surfaces of caries-free subjects. Strains from groups 1 and 2 (ARL, 389 strains; SRS, 358 strains) were previously identified (van Houte et al., 1994) to the genus/species level and belonged to the predominant cultivable flora (PCF). Strains from groups 3 and 4 also belonged to the PCF but were not identified. All strains were placed in one of 4 final pH categories: < 4.2, 4.2-4.4, 4.4-4.6, and > or = 4.6. The main findings were: (1) ARL samples contained many strains with a final pH < 4.2 (mean percentage of 25.7). They included all strains of Lactobacillus and mutans streptococci (MS), most Bifidobacterium strains and non mutans streptococci (non-MS), and about 20% of the Actinomyces strains. By contrast, SRS samples contained far fewer strains with a final pH < 4.2 (mean percentage of 8.4) which were nearly all non-MS. (2) Organisms with a final pH < 4.4 constituted mean percentages of 41.5 and 32.1 for the ARL and SRS samples, respectively. (3) The final pH distribution of strains in samples from coronal surfaces showed a tendency relative to caries activity (group 3 vs. group 4) similar to that for groups 1 and 2. Our findings further support the concept that increased cariogenic conditions are associated with increased proportions of organisms capable of acidogenesis at a low pH and that this shift involves organisms other than the MS and lactobacilli. PMID- 8708130 TI - Enamel defects in the deciduous dentition as a potential predictor of defects in the permanent dentition of 8- and 9-year-old children in fluoridated Cheshire, England. AB - It is claimed that dental fluorosis in both deciduous and permanent teeth is increasing in fluoridated and non-fluoridated communities. What is unclear is whether fluoride-induced enamel opacities in the deciduous dentition are associated with the subsequent appearance of enamel defects in the permanent dentition. The aim of this study was to establish whether a relationship existed between the presence of diffuse enamel defects on the deciduous molars and permanent incisors of schoolchildren who were lifetime residents in an optimally fluoridated community in Cheshire, England. The dentitions of eight-and nine-year old children were examined by two examiners, each unaware of the findings of the other. There was a significant increased risk of diffuse enamel defects in the permanent incisors for those children who presented with diffuse defects on their first deciduous molars (Relative Risk, 1.45; 95% confidence interval, 1.05 to 2.0) or second deciduous molars (Relative Risk, 1.86; 95% confidence interval, 1.36 to 2.54). In light of these findings, it is worth considering the potential of the presence of enamel defects in deciduous molars in children aged 1 to 3 years as a predictor of the future appearance of similar lesions in their permanent incisors. PMID- 8708131 TI - Constant composition dissolution kinetics studies of human dentin. AB - The constant composition (CC) method has been used to study the dissolution kinetics of whole powdered human dentin as a function of calcium phosphate concentration at relative undersaturations with respect to hydroxyapatite (sigma HAP), ranging from +0.8 to -2.8, ionic strength from 0.05 to 0.30 mol/L-1 in sodium chloride or potassium nitrate, pH 4.00 to 5.50, and molar calcium/phosphate ratio in the reaction solutions from 0.05 to 11.1. The results suggest that human dentin behaves as a mixture of at least two calcium phosphate phases, HAP-like and octacalcium phosphate-like, OCP-like. Significant dissolution took place in solutions that were even supersaturated with respect to HAP, and the rates exhibited a striking insensitivity to relative undersaturation, while influenced by ionic strength, pH, and molar calcium/phosphate ratio in the reaction solutions. Although the dissolution was retarded in the presence of magnesium ion, the reaction rate showed the same insensitivity to undersaturation with respect to calcium phosphate. PMID- 8708132 TI - Quantitative analysis of the dentin adhesive interface by Auger spectroscopy. AB - The ultimate success of a dentin adhesive bond is dependent in large part on specific conditions at the interface between the tooth and the adhesive. Most current dentin adhesive systems use some sort of pre-treatment to demineralize the first few microns of the dentin surface, leaving a meshwork of collagen into which the adhesive resin can penetrate, infiltrate, and polymerize. The general hypothesis tested in this experiment was that the penetration and distribution of adhesive resin into the demineralized zone are a function of the conditioner used as a pre-treatment for the adhesive application. Four commercially available adhesive systems were modified to incorporate hydroxyethylthiomethacrylate (HETMA), a sulfur-substituted, traceable analogue of 2-hydroxyethylmethacrylate (HEMA), thereby allowing for a qualitative measurement of the amount and distribution of monomer in the treated dentin substrate by energy-dispersive x ray spectroscopy (EDS) and a quantitative measurement by Auger electron spectroscopy (AES). The dentin pre-treatments investigated were: (1) 10% citric acid/3% ferric chloride, (2) 10% maleic acid, (3) 2.5% nitric acid, and (4) an alcoholic solution of HEMA with a phosphorus acid ester. These pre-treatments were applied to freshly extracted teeth that had been sectioned to expose the dentin and ground to simulate the smeared layer. After the appropriate pre treatment was applied, a 10% (v/v) solution of HETMA in acetone was applied to the surface, followed by the corresponding adhesive resin, which was then polymerized. The samples were then processed for observation by scanning transmission electron microscopy (STEM), AES, and STEM/EDS analysis. The results indicated significant differences in the ability of HETMA to penetrate the dentin surface conditioned by the four pretreatments investigated here. This study also demonstrated that AES and STEM/EDS could be used in a correlative fashion to determine the distribution of HETMA within or adjacent to the treated dentin surface. PMID- 8708133 TI - Resin permeation into acid-conditioned, moist, and dry dentin: a paradigm using water-free adhesive primers. AB - Preservation of the morphological integrity of demineralized dentin collagen in its hydrated state may account for the success observed in wet-bonding procedures. This study investigated the micromorphological differences between moist- and dry-bonding techniques with the use of: (a) Aelitebond, an alcohol based, water-free, single-component dentin adhesive primer system; and (b) a water-free, acetone-based experimental primer similar to the acetone-based, water containing All-Bond 2, a two-component primer system. In the wet groups, acid conditioned dentin surfaces were blotted so that they remained visibly moist prior to bonding. In the dry groups, dentin surfaces were air-dried for 30 sec. Following the bonding procedures, dentin discs in each group were laminated together by means of a chemical-cure resin and processed for scanning electron microscope (SEM) and transmission electron microscope (TEM) examination. Conditioning with 10% H3PO4 for 20 sec. produced complete demineralization of the outer dentin. In the wet groups, banded collagen and interfibrillar spaces could be observed at the surface of the acid-conditioned dentin. Complete wetting of the loosely arranged collagen fibrils by the resin resulted in the formation of a hybrid layer. In the dry groups, only a very thin hybrid layer was observed on the dentin surface, along the walls of the tubules, and along the course of their lateral branches. The absence of banded collagen and interfibrillar spaces within these areas suggested the existence of a collapsed dentin matrix along various liquid-vapor boundaries that restricted resin permeation into the subsurface intertubular matrix, producing an incompletely infiltrated "hybridoid region". PMID- 8708135 TI - Three new rules of life: some additional modest advice for those embarking on a career in dental research. PMID- 8708134 TI - The effect of demineralized bone matrix on the healing of intramembranous bone grafts in rabbit skull defects. AB - A clinical dilemma exists regarding the type of bone that should be used to replace diseased or traumatized osseous tissue. Oral, plastic, and orthopedic surgeons normally implant viable mineralized endochondral (EC) autografts or demineralized EC allografts. A few clinicians have recognized the disadvantages of using EC bone in craniofacial surgery and advocated the replacement of intramembranous (IM) bone with healthy IM bone. However, controversy and uncertainty surround our understanding of these matrices to induce bone formation. Recent studies have advocated the use of other materials with osteoinductive properties, such as demineralized bone matrix (DBM). The proposed delivery system used in this study included IM bone grafts, DBM, and fixation of the IM bone graft. The purpose of this work was to gain further insights into the mechanism of healing of IM bone, in both the presence and the absence of DBM, and to compare the healing of IM bone grafts with that of DBM alone. Critical-sized (10 x 5 mm), full-thickness bony defects in rabbit parietal bone, devoid of periosteum, were filled with IM bone graft (mandible) alone, demineralized cortical bone matrix (DBM) alone, or combined DBM-IM bone graft, or were left unfilled. Histologic changes were examined 14 days later. The IM bone graft healed through IM ossification with no intermediate cartilage stage. DBM and composite DBM-IM healed through an EC ossification with an intermediate cartilage stage. It is hypothesized that the role of the IM graft is to induce neovascularization into the defect site, and that the undifferentiated mesenchymal cells in the perivascular region of the new blood vessels are induced by the bone morphogenetic protein(s) in the DBM into bone-forming cells. PMID- 8708136 TI - The early days of electron microscopy in dental research. PMID- 8708137 TI - Abnormal enamel development in a cystic fibrosis transgenic mouse model. AB - Cystic fibrosis (CF) is a hereditary condition that affects cAMP-regulated chloride channels in epithelial tissues due to a defect in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. Recently, a transgenic CF mouse model was developed at UNC that exhibits no CFTR expression. Interestingly, the CF mouse demonstrates abnormal incisor enamel. Therefore, the purpose of this investigation was to characterize the enamel in this CF mouse model. Incisors from CF and normal mice were evaluated by light microscopy (LM), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). The enamel proteins were examined by amino acid analysis, SDS-PAGE, and Western blot. Gross examination showed that 100% of CF mice had soft, chalky white incisor enamel, while the enamel of normal mice was hard and yellow-brown. LM indicated that the ameloblasts in the CF mice underwent premature degeneration shortly after completion of the secretory phase. The CF mouse enamel appeared to be of relatively normal thickness and showed a prism structure similar to that of normal mouse enamel. However, the CF mouse enamel crystallites appeared to have a rough granular surface compared with normal enamel. SDS-PAGE indicated that mature CF enamel retained low-molecular-weight material (approximately 20 kDa), whereas normal mature enamel did not. This low-molecular-weight material cross reacted with anti-amelogenin antibodies in Western blot analysis. This investigation shows that abnormal CFTR expression in the mouse results in developmental abnormalities in the incisor enamel. Although further investigation is required to determine the mechanism leading to abnormal enamel formation, the CF mouse provides a potentially useful animal model for investigating aberrant enamel development. PMID- 8708138 TI - Highly virulent strains of herpes simplex virus fail to kill mice following infection via gingival route. AB - Virulence of herpes simplex virus (HSV) in mice has been demonstrated to be dependent on the site of infection. In this experiment, pathogenesis of HSV was studied in 2 different routes of infection in a mouse model system. When BALB/c mice were infected with 5 x 10(3) plaque-forming units (PFU) of virulent HSV type 1 Miyama GC+ strain (HSV-1-GC+) intraperitoneally, all mice were dead in 6 to 9 days. HSV-1-GC+ was recovered from organs such as the cerebrum, cerebellum, brainstem, and spleen 2 to 5 days after infection, but not from other organs such as trigeminal ganglia. However, if mice were infected in the maxillary gingiva with 1.0 x 10(7) PFU of HSV-1-GC+, all mice survived. HSV-1-GC+ was recovered from the trigeminal ganglia and brainstem 2 to 5 days after infection, but not from other organs tested. When mice were infected in maxillary gingiva with HSV-1 GC+, followed by the intraperitoneal injection of 6 mg of cyclophosphamide 72 hrs after virus infection, all mice were dead within days. Immunofluorescent and hematoxylin-eosin staining of gingival tissue sections revealed that when mice were infected in maxillary gingiva with HSV-1-GC+, 3 times as many gamma delta T cells and 5 times as many polymorphonuclear cells can be detected in sections of maxillary gingiva when compared with non-infected mice. These data show that the gingiva of mice is considerably more resistant to infection with HSV, compared with the peritoneal cavity, and suggest the possible presence of an oral defense mechanism which might be different from that in the peritoneal cavity. PMID- 8708139 TI - The flow rate of unstimulated human labial gland saliva. AB - Although the minor salivary glands contribute an important fraction of the resting flow of saliva, which may be particularly relevant in xerostomia, there is currently no precise method of measuring the flow from individual glands or recording the number of active glands in a given area. These experiments were devised to test a photographic method of assaying labial gland saliva and to see whether subjects of different ages had differing flow rates and/or numbers of glands. Unstimulated saliva secretion from labial salivary glands was measured in healthy non-medicated subjects aged 20 to 55 years by a photographic method. The number of glands in a unit area, the individual gland flow rates, and the flow rates per unit area were assessed. There was a high correlation between flow rate per unit area as measured by this method and measurements from weighing filter paper before and after absorption of the saliva droplets when the latter method could be used. The mean flow rate for 22 subjects was 2.35 +/- 1.65 microL/min over the whole area photographed. Subjects with higher flow rates had both more active glands and a higher rate of secretion from individual glands. There was no correlation between either number of active glands in a unit area or of flow rate and age in the age range represented by these subjects. PMID- 8708140 TI - Comparative study of the chemotactic responses of periodontal ligament cells and gingival fibroblasts to polypeptide growth factors. AB - Selective recruitment of periodontal ligament cells to a previously exposed root surface is believed to enhance periodontal regeneration. It has been hypothesized that competition from gingival fibroblasts may reduce the potential of periodontal regeneration. We compared the migratory responses of PDL cells and gingival fibroblasts to a variety of biologicals. Parallel experiments designed to examine the directed migration responses of both periodontal ligament cells (PDL cells) and gingival fibroblasts (GF) isolated from the same donors were conducted using Platelet Derived Growth Factor (PDGF), Insulin Like Growth Factor I, -II (IGF-I, -II), Epidermal Growth Factor (EGF), Transforming Growth Factor beta (TGF-beta), and the chemotactic factor derived from the conditioned culture media of PDL cells (termed PDL-CTX) as attractants. Both PDL cells and GF exhibited dose-dependent migratory responses when challenged with PDGF, IGF-I, IGF-II, EGF, and TGF-beta. However, when these cells were challenged with PDL CTX, only PDL cells migrated in a specific dose-dependent manner, while GF were refractive to PDL-CTX stimulation. Additionally, concentrated conditioned culture media from cultures of gingival fibroblasts did not stimulate PDL cell migratory responses. In other experiments, antibody directed against PDGF, FGF, TGF-beta, IGF-I, IGF-II, NGF, and EGF did not inhibit the PDL-CTX-elicited response in PDL cells. Previous studies have suggested that success of periodontal therapy depends on the specific attachment, migration, and proliferation of selected periodontal ligament cells. The data presented in this manuscript suggest that both PDL cells and gingival fibroblasts respond to a multitude of growth factors. PDL-CTX was found to be PDL-cell-specific for directed migration. Thus, we conclude that any biological therapeutic regime for periodontal regeneration should include PDL-cell-specific agents. PMID- 8708141 TI - Periodontal ligament cells are chemotactic to fibroblast collagenase. AB - Periodontal ligament (PDL) cell motility and the passage of PDL cells along a root surface are important components of tissue remodeling during periodontal regeneration. Proteolytic enzymes, including fibroblast collagenase, have been demonstrated to play an important role in tissue remodeling. Previous studies have shown that PDL cells chemotactically respond to a variety of matrix and growth factors. We therefore studied the effects of type I collagen fragments and fibroblast collagenase on PDL cell migration, since PDL cells have been shown to adhere preferentially to partially demineralized root surfaces with exposed type I collagen. Gingival epithelial cells were used as a control cell population. We report that PDL cells but not gingival epithelial cells preferentially migrate in a dose-dependent manner to both fibroblast collagenase and to type I collagen degradation products. Epithelial cell migration to fibroblast collagenase and type I collagen fragments was observed. Antibody to type I collagen inhibited the type I collagen fragment-mediated migration. Collagenase pre-treatment of PDL cells enhanced PDL cell migration to type I collagen fragments. In other assays, enzyme inhibitors were shown to decrease the collagenase-mediated PDL cell motility. Epithelial cells were shown to migrate preferentially to 92-kDa type IV collagenase and type IV collagen degradation products. Antibody to type IV collagen inhibited type IV collagen-induced epithelial cell migration. Taken together, these data suggest a role for collagenase in the fine control of PDL cell migration in tissue remodeling during periodontal regeneration. PMID- 8708142 TI - Characteristics of general dentists participating in home study courses. AB - With an increasing number of states requiring continuing dental education (CDE) for license renewal, dentists are opting for different ways of earning these credit hours beyond the traditional didactic presentation. One alternative is a home study correspondence course. Little is known about the type of dentists who participate in these home study courses. This study examines characteristics of 507 general dentists in Ohio who were required to submit information concerning their CDE courses and credit hours during the biennium, 1992-93. Twenty-five percent of the general dentists took at least one home study course, and approximately five percent of those dentists participating in any home study earned all their credit hours this way. A logistic regression model indicated that home study users are less likely to be ADA members and less likely to leave the state for any CDE. This study suggests that the current methods of home study courses may provide continuing education opportunities for dentists who otherwise might not be reached by more traditional ways of CDE. PMID- 8708144 TI - Values of selected qualitative methods for research, education, and policy. PMID- 8708145 TI - Combining qualitative and quantitative research approaches in understanding pain. PMID- 8708143 TI - Potential effects of opportunity cost on dental school patients. AB - Predoctoral clinic fees are set below those of private practices to attract patients. Treatment in a predoctoral clinic requires more and longer appointments and patients spend more time waiting to be seen than in a private practice. This time has a value that economists call opportunity cost, the value of the next best alternative (the hourly wage for an employed patient). A model comparing total cost (fee plus opportunity cost) of treatment in dental schools and private practices for insured and uninsured patients is presented. As patient income increases, the total cost of some treatment plans becomes higher in dental schools than in private practices. To insured patients, however, the out-of pocket cost of a treatment plan depends on copayments rather than fees. This makes predoctoral clinics less competitive. Considering opportunity cost further increases dental schools' lack of competitiveness. For insured patients, more treatment plans will have a lower total cost in private practice. Clinic directors must realize that if opportunity costs are not reduced, the market may dictate that fees be lowered to remain competitive with private practitioners. They are in competition for patients based on total cost, and efficiencies that reduce opportunity cost may increase revenue and attract patients. PMID- 8708146 TI - Understanding and utilizing qualitative research. PMID- 8708147 TI - Update on Medicare graduate medical education law, regulations, and legislative proposals. PMID- 8708148 TI - In vitro interleukin-5 (IL-5) production by peripheral blood mononuclear cells from patients with drug hypersensitivity. AB - It has been shown that CD4+ lymphocytes and eosinophils pathologically infiltrate the dermis in skin eruptions due to hypersensitivity to the administration of certain drugs associated with peripheral eosinophilia. However, the mechanisms involved in drug related skin eruptions with eosinophilia are largely unknown. There are several methods of diagnosing drug hypersensitivity, but no single in vitro method is available for the detection of the sensitizing drug. In order to elicit the mechanism of drug related skin eruption with eosinophilia and to establish effective methods for diagnosing drug hypersensitivity, we studied in vitro IL-5 production by peripheral mononuclear cells (PBMC) from patients with drug related skin eruptions (n = 8). Significant increase in IL-5 from exposure to drug alone (n = 3) and drug with PHA (n = 3) were noted, whereas PBMC from normal subjects (n = 7) exhibited no such stimulation. Six of the eight cases showed a significant decrease in the number of peripheral blood eosinophils when the skin eruptions healed compared with those at the onset. Histopathological features revealed mild to moderate infiltration of eosinophils. Immunohistopathologically, infiltration of CD4+ and CD8+ lymphocytes and CD4+ cells were predominant within the dermis of drug related skin eruption sites in all cases. This study suggested that the IL-5 production of sensitized mononuclear cells might be a critical mediator in drug hypersensitivity with eosinophilia and an important diagnostic marker. PMID- 8708149 TI - Serum levels of eosinophil cationic protein reflect the state of in vitro degranulation of blood hypodense eosinophils in atopic dermatitis. AB - In patients with atopic dermatitis (AD), serum levels of eosinophil cationic protein (ECP) have been shown to be a good reflector of disease severity. To elucidate what serum levels of ECP actually reflect, ECP levels in serum and plasma and cytological aspects of blood eosinophils were examined in AD patients (n = 27) and compared to healthy subjects (n = 12). Significantly elevated levels of serum ECP were noted in AD patients, while plasma ECP were uniformly recorded at nadir levels in both AD patients and normal subjects. In addition to blood eosinophilia, AD patients had significantly increased numbers of hypodense eosinophils (HEo) with morphological characteristics consistent with an activated state. Serum ECP levels strongly correlated with HEo numbers rather than with total eosinophil counts. These results indicate that elevated levels of serum ECP may be a consequence of in vitro degranulation of "activated" HEo, not of ECP supplementation from lesional skin. In addition, the dynamic correlations of eosinophil-associated parameters (total eosinophil counts, HEo numbers, and serum ECP levels) with AD severity suggest that inflammatory events in lesional skin may be involved in causing not only eosinophilopoiesis in bone marrow, but also development of HEo in the periphery, whose degree in turn may be mirrored in the levels of serum ECP in vitro. PMID- 8708150 TI - Distribution of lectin UEA-I in trichilemmal carcinoma, squamous cell carcinoma, and other epithelial tumors of the skin. AB - Trichilemmal carcinoma (TLC) is a cutaneous appendage tumor. In some cases, the histological distinction between TLC and squamous cell carcinoma (SCC) is difficult with hematoxylin & eosin staining. In the present study, in order to establish a simple and reliable method for distinguishing between TLC and SCC, we examined the lectin Ulex europaeus agglutinin-I (UEA-I)-binding patterns of TLC (4 cases), SCC (10 cases), malignant proliferating trichilemmal cyst (MPTC) (3 cases), Bowen's disease (4 cases), Bowen's carcinoma (4 cases), actinic keratosis (4 cases), basal cell carcinoma (8 cases), and eccrine porocarcinoma (1 case). In normal skin samples, UEA-I was strongly positive in the outer root sheath cells of the hair follicle and weakly positive in the granular layers of the epidermis. UEA-I stained the tumor cells of TLC clearly and some MPTC and Bowen's disease cells weakly. This result indicates that histochemical staining with UEA-I is a simple and useful method for distinguishing between TLC and SCC of the skin. PMID- 8708151 TI - Comparison of the effects of intralesional interferon alfa-2a, 2b and the combination of 2a and 2b in the treatment of basal cell carcinoma. AB - Interferons (IFN), especially alfa-2a and 2b, are potent inhibitors of normal and malignant cell growth and effective choices in the treatment of basal cell carcinoma (BCC). The aim of the study was to evaluate the effectiveness of the IFN alfa-2a and 2b, and whether this effect may be increased by their combination. A total of 45 patients with histopathologically confirmed BCC were divided into three groups, equally. Groups 1,2 and 3 were treated with intralesional IFN alfa-2a, 2b, and the combination of 2a and 2b, respectively. Lesions were injected three times weekly in Groups 1 and 2 (total ten injections). IFN alfa-2a and 2b were injected alternately in Group 3 (5 IFN alfa 2a and 5 IFN alfa-2b). Eight weeks after completion of therapy, cytologic specimens were taken, and all cases were evaluated clinically and cytologically. Ten (66.6%) patients had a complete response (CR) and 5 (33.3%) had a partial response (PR) in Group 1. Patients in Group 2 showed 10 (66.6%) CR, 4 (26.6%) PR, and 1 (6.6%) non response. In Group 3, 11 (73.3%) patients had CR and 4 (26.6%) had PR. Follow-up periods ranged from 6-24 months, and no recurrences have been observed. These results suggest that IFN alfa-2a, 2b, and the combination of 2a and 2b appear to be effective choices for the treatment of BCC; the effectiveness is not increased by their combination. PMID- 8708152 TI - Paraneoplastic pemphigus: report of a case. AB - A 56-year-old male with chronic lymphocytic leukemia developed extensive erosive mucocutaneous lesions with histologic acantholysis. Immunopathologic studies showed IgG deposition at the intercellular space, C3 deposition at both the intercellular space and the dermo-epidermal junction, and reactivity of the serum to rat urinary bladder epithelium. Autoantibodies in the serum to human epidermal proteins of 210 kD and 190 kD were shown by Western blotting and to proteins of 250 kD, 210 kD, and 190 kD by immunoprecipitation. All these data suggest the diagnosis of paraneoplastic pemphigus. Repeated plasmapheresis resulted in re epithelialization of the mucocutaneous lesions and reduction in antibody titer from 1:1280 to 1:20. Although this mucocutaneous disease was established as a new autoimmune bullous disease by Anhalt et al. (1990), cases have rarely been reported from Japan. The present patient demonstrates the major characteristics of paraneoplastic pemphigus. PMID- 8708153 TI - Subcutaneous granuloma annulare with Hodgkin's disease. AB - We report an 85-year-old woman with subcutaneous granuloma annulare. Three nodules developed on her scalp and sacral regions and gradually increased with tenderness. Histologically, a biopsy specimen of the nodule was interpreted as subcutaneous granuloma annulare because palisading granuloma was present in the dermis and subcutaneous fatty tissue. Laboratory tests, including RA, showed no remarkable changes except for hyperglycemia and glycosuria. These nodules spontaneously regressed without medication. One year later, she noted swelling of some lymph nodules in the left axillary region. These were diagnosed as Hodgkin's disease (mixed cellularity type). She was treated with chemotherapy, but died after two months. Subcutaneous granuloma annulare which occurs suddenly in the elderly may be a dermadrome of malignant blood disease. PMID- 8708154 TI - Extramammary Paget's disease with distant skin metastasis. AB - We report a 68-year-old Japanese man with invasive extramammary Paget's disease (EPD) which started in the left inguinal area and enlarged to include the scrotal skin, penis, and pubic area, resulting in lymph edema of the entire left leg. Histopathological examination of one nodule revealed an intraepidermal and intradermal arrangement of solid tumor cells with characteristic pale cytoplasm. The tumor was found to have disseminated to regional and systemic lymph nodes, lungs, bones, and the left adrenal gland. In addition, the patient had a metastatic cutaneous nodule on the scalp. Histology of the skin metastasis showed that the proliferation of Paget cells was confined to the dermis. This type of metastatic spread of EPD is very rare; only two other apparent cases in Japan have been reported. PMID- 8708155 TI - Inflammatory linear verrucose epidermal nevus coexisting with lichen amyloidosus. AB - The clinical and histopathological findings in a case of inflammatory linear verrucose epidermal nevus coexisting with lichen amyloidosus are presented. A 33 year-old woman had had linear verrucose lesions which were histopathologically compatible with ILVEN, and with which no amyloid deposits were associated, on her left lower leg for 30 years. She had noticed keratotic papules around the verrucose lesions on the left leg for the last 3 years. Histopathology of these papules revealed amyloid deposits in the upper dermis under the hyperkeratotic and acanthotic epidermis. Topical application of DMSO liniment with dexamethasone successfully treated the ILVEN. This remission of both lesions and itching also alleviated the keratotic papules on the left leg. PMID- 8708156 TI - Two cases of pachydermodactyly. AB - We report two cases of pachydermodactyly. Case 1 was a 16-year-old girl who complained of asymptomatic, bulbous, firm swellings which developed insidiously on both sides of the proximal interphalangeal (PIP) joint of her right middle finger. Case 2 was a 14-year-old boy with similar lesions on the sides of the PIP joints of the index and middle fingers of both hands. They both had histories of mild, repetitive mechanical trauma of the fingers. Radiologic findings showed soft tissue swellings without any bony or articular abnormalities. Histopathologic findings from the bulbous swellings revealed marked hyperkeratosis, slight epidermal hyperplasia, and a markedly thickened dermis with a deposition of mucinous material among the collagen fibers. Ultrastructural examinations of both cases showed decreased diameters of collagen fibrils. The lesions temporarily improved with intralesional injection of triamcinolone acetonide. Pachydermodactyly is more commonly found in boys and the affected fingers are more numerous in boys. Both of the present cases had the habit of rubbing and gripping their fingers unconsciously. Mechanical trauma of the fingers around puberty may play an important role in pachydermodactyly. PMID- 8708157 TI - Hypopigmented mycosis fungoides in a white female. AB - A rare case of a young Caucasian female with hypopigmented mycosis fungoides is described. We reviewed and discussed the literature. PMID- 8708158 TI - Multiple milia localized to the vulva. AB - Milia are most commonly observed on the cheeks and eyelids. We studied a case of multiple milia localized to the vulva in a 64-year-old female. A review of English and Japanese literature for the last 20 years failed to uncover any reports of milia limited to this area. We describe this case in detail and provide a short review of the literature on cysts of the genitalia. PMID- 8708159 TI - Depigmented macula as an initial manifestation of extramammary Paget's disease. PMID- 8708161 TI - Varicella in an 81-year-old woman. PMID- 8708160 TI - Recurrent annular erythema in Sjogren's/lupus erythematosus overlap syndrome: an additional case from Korea. PMID- 8708162 TI - Pulmonary oxygen consumption. PMID- 8708163 TI - Relationship between pulmonary oxygen consumption, lung inflammation, and calculated venous admixture in patients with acute lung injury. AB - OBJECTIVE: To determine in patients with acute lung injury whether increased pulmonary oxygen consumption (VO2pulm), computed as the difference between oxygen consumption measured by indirect calorimetry (VO2meas) and calculated by the reverse Fick method (VO2Fick), would: (1) correlate with the degree of lung inflammation assessed by bronchoalveolar lavage (BAL); (2) lead to an overestimation of calculated venous admixture (Qva/Qt). DESIGN: Prospective study. SETTING: University hospital, medical intensive care unit. INTERVENTION: None. MEASUREMENTS AND RESULTS: In nine mechanically ventilated patients with acute lung injury (Apache II 12 +/- 5, lung injury score 2 +/- 0.6, mean +/- SD), whole-body VO2 (VO2wb) was determined simultaneously by indirect calorimetry and the reverse Fick technique, after which BAL was immediately performed. VO2meas was significantly higher than VO2Fick (128 +/- 24 and 102 +/- 18 ml/min per m2, respectively, p < 0.001). Median VO2pulm was 25.3 ml/min per m2 (range 1.98 51.5), thus representing 19 +/- 11% of VO2wb. Total BAL cellularity was increased in all patients (median 47, range 24-200 x 10(4)/ml), as was the total polymorphonuclear (PMN) count (median 78 range 5-93 x 10(4)/ml). Macrophage counts were in the normal range. There were raised BAL levels of interleukin-6 (IL-6) (median 945, range 23-1800 ng/ml) and elastase (median 391, range 5-949 ng/ml). Median protein levels were 270 micrograms/ml (range 50-505). There was no correlation between VO2pulm and BAL cellularity, PMNs, elastase, IL-6, or protein. Qva/Qt was 31.7 +/- 8%. Qva/Qt, corrected for the presence of VO2pulm, (Qva/Qtcorr), was 30.3 +/- 8% (P < 0.01 vs Qva/Qt), a 4.2% overestimation due to VO2plum. There was no correlation between Qva/Qt or Qva/Qtcorr and VO2pulm. CONCLUSIONS: In mechanically ventilated patients with acute lung injury, VO2pulm was increased and led to a 19% underestimation of VO2wb determined by the reverse Fick method, as well as to a 4.2% overestimation of calculated Qva/Qt. Lung inflammatory activity was increased, as assessed by BAL cellularity, IL-6 and elastase levels. However, there was no correlation between VO2pulm and the intensity of pulmonary inflammation. PMID- 8708164 TI - Intercurrent complications in chronic alcoholic men admitted to the intensive care unit following trauma. AB - OBJECTIVE: A chronic alcoholic group following trauma was investigated to determine whether their ICU stay was longer than that of a non-alcoholic group and whether their intercurrent complication rate was increased. DESIGN: Prospective study. SETTING: An intensive care unit. PATIENTS: A total of 102 polytraumatized patients were transferred to the ICU after admission to the emergency room and after surgical treatment. Of these patients 69 were chronic alcoholics and 33 were allocated to the non-alcoholic group. The chronic alcoholic group. met the DSM-III-R and ICD-10 criteria for alcohol dependence or chronic alcohol abuse/harmful use. The daily ethanol intake in these patients was > or = 60 g. Diagnostic indicators included an alcoholism-related questionnaire (CAGE), conventional laboratory markers and carbohydrate-deficient transferrin. MEASUREMENT AND RESULTS: Major intercurrent complications such as alcohol withdrawal syndrome (AWS), pneumonia, cardiac complications and bleeding disorders were documented and defined according to internationally accepted criteria. Patients did not differ significantly between groups regarding age, TRISS and APACHE score on admission. The rate of major intercurrent complications was 196% in the chronic alcoholic vs 70% in the non-alcoholic group (p = 0.0001). Because of the increased intercurrent complication rate, the ICU stay was significantly prolonged in the chronic-alcoholic group by a median period of 9 days. CONCLUSIONS: Chronic alcoholics are reported to have an increased risk of morbidity and mortality. However, to our knowledge, nothing is known about the morbidity and mortality of chronic alcoholics in intensive care units following trauma. Since chronic alcoholics in the ICU develop more major complications with a significantly prolonged ICU stay following trauma than non-alcoholics, it seems reasonable to intensify research to identify chronic alcoholics and to prevent alcohol-related complications. PMID- 8708165 TI - Increased oxygen consumption after cardiac surgery is associated with the inflammatory response to endotoxemia. AB - OBJECTIVE: The aim of this study was to determine whether the increase in post operative oxygen consumption (delta VO2) in cardiac surgery patients is related to endotoxemia and subsequent cytokine release and whether delta VO2 can be used as a parameter of post-perfusion syndrome. DESIGN: Prospective study. SETTING: Operating room and intensive care unit of a university hospital. PATIENTS: Twenty one consecutive male patients undergoing elective coronary artery bypass surgery without major organ dysfunction and not receiving corticosteroids. MEASUREMENTS AND RESULTS: Plasma levels of endotoxin, tumor necrosis factor (TNF) and interleukin-6 (IL-6) were measured before, during and for 18 h after cardiac surgery. Oxygen consumption, haemodynamics, the use of IV fluids and dopamine, body temperature and the time of extubation were also measured. Measurements from patients with high delta VO2 (> or = median value of the entire group) were compared with measurements from patients with low delta VO2 (< median). Patients with high delta VO2 had higher levels of circulating endotoxin (P = 0.004), TNF (P = 0.04) and IL-6 (P = 0.009) received more IV fluids and dopamine while in the ICU, and were extubated later than patients with low delta VO2. Several hours after delta VO2 the patient's body temperature rose. Forward stepwise regression analysis showed that circulating endotoxin and TNF explained 50% of the variability of delta VO2. CONCLUSIONS: This study demonstrates that patients with high post operative oxygen consumption after elective cardiac surgery have higher circulating levels of endotoxin, TNF and IL-6 and also have more symptoms of post perfusion syndrome. Early detection of high VO2 might be used as a clinical signal to improve circulation in order to meet the high oxygen demand of inflammation. In addition, continuous measurement of VO2 provides us with a clinical parameter of inflammation in interventional studies aiming at a reduction of endotoxemia or circulating cytokines. PMID- 8708166 TI - Motility agents for the placement of weighted and unweighted feeding tubes in critically ill patients. AB - OBJECTIVE: To determine if successful attempts at feeding tube placement into the small bowel could be increased with the use of a weighted end or by pre-treatment with a drug to increase gastric motility. DESIGN: A prospective randomized control study, double blinded for a drug study drug, in a population of critically ill patients. SETTING: A 635-bed acute care hospital in Philadelphia, Pennsylvania. PATIENTS: Eighty-three patients in the critical care setting randomized into four groups receiving either parenteral normal saline (NS) 100 cc, erythromycin (EMY) 200 mg, or metoclopramide (MET), 10 mg, 30 min prior to attempted tube placement with either a weighted (WEI) (57 patients) or unweighted tube (UNW) (26 patients). RESULTS: When analyzed for number of attempts prior to successful tube placement into the stomach there was a significant difference between the unweighted and weighted groups: 2.08 +/- 1.03 attempts vs 1.51 +/- 0.94, P < or = 0.015. Duodenal migration at 24 h was demonstrated in three patients in the NS/UNW group and in two patients in the NS/WEI group as compared to no patients in either the EMY/WEI or the MET/WEI groups (p < or = 0.025, Fisher's exact test). CONCLUSIONS: These data demonstrate that the use of weighted feeding tubes decreases the number of attempts required to achieve gastric intubation, but that motility agents given prior to tube insertion do not augment advancement of the feeding tube beyond the stomach and may in fact hinder placement into the duodenum. PMID- 8708167 TI - The effects of pentoxifylline on circulating adhesion molecules in critically ill patients with acute renal failure treated by continuous veno-venous hemofiltration. AB - OBJECTIVE: Circulating adhesion molecules appear to be excellent markers of endothelial activation in critically ill patients. Pentoxifylline (PTX) may limit sequelae of inflammation and subsequent endothelial activation by various mechanisms. The influence of PTX on the plasma levels of soluble adhesion molecules in critically ill patients undergoing continuous veno-venous hemofiltration (CVVH) was studied. DESIGN: Prospective, randomized, blinded study. SETTING: Clinical investigation in the surgical intensive care unit of a university hospital. PATIENTS AND PARTICIPANTS: Fourteen consecutive patients suffering from acute renal failure (ARF) with postoperative complications who received continuous pentoxifylline (CVVH-PTX) i.v. were compared with 14 patients with ARF who did not receive PTX (CVVH control group). INTERVENTIONS: Pump-driven CVVH was carried out with a blood flow ranging from 120 to 150 ml/min. All patients received fentanyl and midazolam continuously and were on mechanical ventilation. PTX (300 mg) was given as a loading dose, followed by continuous infusion of 1.2 mg/kg per h for the next 5 days. MEASUREMENTS AND RESULTS: From arterial blood samples, plasma levels of soluble adhesion molecules (endothelial leukocyte adhesion molecules [sELAM-1], and intercellular adhesion molecule-1 [sICAM-1], vascular cell adhesion molecule-1 [sVCAM-1], and P-selectin granule membrane protein [sGMP-140] were measured using enzyme-linked immuno-sorbent assays (ELISA). Measurements were carried out before the start of CVVH to establish baseline values and continued during the next 5 days. MAIN RESULTS: Eleven of the CVVH-PTX patients and 8 of the CVVH control patients survived during the investigation period. In the CVVH-PTX patients 2.4 +/- 0.3 g/day of PTX was given. At baseline, plasma levels of sELAM-1, sICAM-1, and sVCAM-1 were markedly higher than normal in both groups. In the CVVH control patients, all measured soluble adhesion molecules increased further during the study period (sELAM-1 from 90 +/- 22 to 134 +/- 30 ng/ml; sICAM-1 from 958 +/- 173 to 1460 +/- 209 ng/ml; sVCAM-1 from 1100 +/- 188 to 1804 ng/ml; sGM-140 from 499 +/- 102 to 688 +/- 121 ng/ml) (p < 0.05), whereas in the PTX-treated CVVH patients, plasma levels of all soluble adhesion molecules remained almost unchanged. The PaO2/FIO2 increased in the PTX-treated patients (from 209 +/- 67 to 282 +/- 58 mmHg) and remained almost unchanged in the CVVH control patients. CONCLUSION: Leukocyte/endothelial interactions play an important role in the inflammatory process. Circulating adhesion molecules may serve as markers of the extent of inflammation. Continuous i.v. administration of PTX was successful in blunting the increase of soluble adhesion molecules in critically ill patients undergoing CVVH. Whether these effects result from improved circulation at the microcirculatory level or from (direct or indirect) beneficial effects on endothelial cells warrants further controlled studies. PMID- 8708168 TI - Semi-continuous versus injectate cardiac output measurement in intensive care patients after cardiac surgery. AB - OBJECTIVE: Commercially available semi-continuous cardiac output (SCCO) monitoring systems are based on the pulsed warm thermodilution technique. There is evidence that SCCO fails to correlate with standard intermittent bolus cardiac output (ICO) in clinical situations with thermal instability in the pulmonary artery. Furthermore, ventilation may potentially influence thermodilution measurements by enhanced respiratory variations in pulmonary artery blood temperature and by cyclic changes in venous return. Therefore, we evaluated the correlation, accuracy and precision of SCCO versus ICO measurements before and after extubation. DESIGN: Prospective cohort study. SETTING: Intensive care unit (ICU) of a university hospital. PATIENTS AND PARTICIPANTS: 22 cardiac surgical ICU patients. INTERVENTIONS: None. MEASUREMENTS AND RESULTS: SCCO and ICO data were obtained at nine postoperative time points while the patients were on controlled mechanical ventilation. Further sets of measurements were taken during the weaning phase 20 min before extubation, and 5 min, 20 min and 1 h after extubation. SCCO and ICO measurements yielded 286 data pairs with a range of 1.8 9.9 l/min for SCCO and 1.9-9.8 l/min for ICO. The correlation between SCCO and ICO was highly significant (r = 0.92; p < 0.01), accompanied by a bias of -0.052 l/min and a precision of 0.56 l/min. Correlation, accuracy and precision were not influenced by the mode of respiration. CONCLUSIONS: Our results demonstrate excellent correlation, accuracy and precision between SCCO and ICO measurements in postoperative cardiac surgical ICU patients. We conclude that SCCO monitoring offers a reliable clinical method of cardiac output monitoring in ICU patients following cardiac surgery. PMID- 8708170 TI - Bladder epithelial oxygen tension--a new means of monitoring regional perfusion? Preliminary study in a model of exsanguination/fluid repletion. AB - OBJECTIVE: To assess whether monitoring of bladder epithelial oxygen tension (BEOT) would provide an indication of regional (renal) organ perfusion in an exsanguination/fluid repletion animal model. DESIGN: Prospective non-randomized laboratory study. SETTING: Research laboratory. INTERVENTIONS: Eight anaesthetised, spontaneously breathing Sprague-Dawley male rats weighing approximately 200 g were instrumented. They received 1-ml aliquots of fluid until no further haemodynamic improvement was seen, followed by removal of 1-ml aliquots of blood until renal blood flow fell by 50%. The animal was then resuscitated with repeated 1 to 2-ml aliquots of fluid until no further improvement was achieved and, finally, progressively exsanguinated to cardiovascular collapse. MEASUREMENTS AND RESULTS: A continuous Clark-type oxygen electrode lying in contact with the inside wall of the bladder measured changes in BEOT during these exsanguination and fluid repletion manoeuvres. Changes in BEOT closely mirrored both systemic (blood pressure and aortic blood flow) and regional (renal blood flow) haemodynamic changes. A direct correlation existed between percentage change in BEOT and base deficit, and an indirect correlation was seen with arterial oxygen tension. CONCLUSIONS: Measurement of BEOT may be a useful and relatively non-invasive means of monitoring regional organ perfusion. Further studies are warranted. PMID- 8708169 TI - Effect of nasogastric suction and ranitidine on the calculated gastric intramucosal pH. AB - OBJECTIVE: To study the effect of nasogastric suction and ranitidine on the determination of gastric intramucosal pH (pHi). DESIGN: Prospective study. SETTING: Clinical research unit at a university hospital intensive care department. SUBJECTS: 12 healthy volunteers. INTERVENTIONS: After a 2-h measurement control period a tonometer was connected to nasogastric suction for 2 h, and thereafter ranitidine was given intravenously and gastric pHi measured. MEASUREMENTS AND RESULTS: During each 2-h measurement period gastric PCO2, gastric pHi, and pH gap were determined every 30 min. Luminal pH was measured after insertion of tonometer and at the end of each study period. Neither nasogastric suction nor ranitidine had an effect on the coefficient of variation for either gastric PCO2 or pHi. Compared to control and nasogastric suction periods, after ranitidine mean gastric pHi was higher (control 7.22 +/- 0.08; nasogastric suction 7.23 +/- 0.07; after ranitidine 7.31 +/- 0.06, p < 0.001) mean gastric PCO2 lower (control 6.4 +/- 1.3; nasogastric suction 6.5 +/- 1.3; after ranitidine 5.3 +/- 0.9, p < 0.001) and pH gap lower (control 0.18 +/- 0.08; nasogastric suction 0.17 +/- 0.05; after ranitidine 0.09 +/- 0.06, p < 0.01). Luminal pH increased after ranitidine in each subject. CONCLUSIONS: H2 blockers have no effect on the reproducibility of gastric pHi measurements, but the use of H2 blockers modifies the normal values for gastric pHi in healthy subjects. PMID- 8708171 TI - Oxygenation remains unaffected by increased inspiration-to-expiration ratio but impairs hemodynamics in surfactant-depleted piglets. AB - OBJECTIVES: Prolongation of inspiratory time is used to reduce lung injury in mechanical ventilation. The aim of this study was to isolate the effects of inspiratory time on airway pressure, gas exchange, and hemodynamics, while ventilatory frequency, tidal volume, and mean airway pressure were kept constant. DESIGN: Randomized experimental trial. SETTING: Experimental laboratory of a University Department of Anesthesiology and Intensive Care. ANIMALS: Twelve anesthetised piglets. INTERVENTIONS: After lavage the reference setting was pressure-controlled ventilation with a decelerating flow; I:E was 1:1, and PEEP was set to 75% of the inflection point pressure level. The I:E ratios of 1.5:1, 2.3:1, and 4:1 were applied randomly. Under open lung conditions, mean airway pressure was kept constant by reduction of external PEEP. MEASUREMENTS AND RESULTS: Gas exchange, airway pressures, hemodynamics, functional residual capacity (SF6 tracer), and intrathoracic fluid volumes (double indicator dilution) were measured. Compared to the I:E of 1:1, PaCO2 was 8% lower, with I:E 2.3:1 and 4:1 (p < or = 0.01) while PaO2 remained unchanged. The decrease in inspiratory airway pressure with increased inspiratory time was due to the response of the pressure-regulated volume-controlled mode to an increased I:E ratio. Stroke index and right ventricular ejection fraction were depressed at higher I:E ratios (SI by 18% at 2.3:1, 20% at 4:1; RVEF by 10% at 2.3:1, 13% at 4:1; p < or = 0.05). CONCLUSION: Under open lung conditions with an increased I:E ratio, oxygenation remained unaffected while hemodynamics were impaired. PMID- 8708172 TI - A stable model of respiratory distress by small injections of oleic acid in pigs. AB - OBJECTIVE: Development of a stable model of respiratory distress in pigs with oleic acid, fulfilling clinical criteria of the adult respiratory distress syndrome (ARDS). DESIGN: Eight pigs (9.1 +/- 0.7 kg) were anesthetized with pentobarbital, paralyzed with tubocurarine and mechanically ventilated with an FIO2 of 0.6, an I:E ratio of 2:3 and a PEEP of 0.2 kPa. Oleic acid (dissolved 1:1 in 96% alcohol) was administered in a series of multiple injections of 0.1 ml until PaO2 was lower than 8 kPa. MEASUREMENTS AND RESULTS: Careful titration of the oleic acid injections on guidance of the PaO2 established a reproducible respiratory distress (PaO2 = 7.3 +/- 0.8 kPa), in which gas exchange and hemodynamic variables were stable for at least 4 h. The number of oleic acid injections (22 +/- 11, mean and SD) varied between the animals. CONCLUSIONS: With the use of multiple injections of oleic acid, a stable model of early respiratory distress in pigs can be achieved, in spite of individual differences in sensitivity. Such a stable model allows for a diversity of studies on early respiratory distress. PMID- 8708173 TI - Effect of N-nitroso-N-methylurethane on gas exchange, lung compliance, and surfactant function of rabbits. AB - OBJECTIVES: To define the effect of N-nitroso-N-methyl-urethane (NNNMU) on pulmonary gas exchange, compliance and the biochemical and functional properties of the lung surfactant system. DESIGN: Four days after inducing lung injury, gas exchange and pulmonary compliance were studied and a bronchoalveolar lavage was taken. SETTING: Experimental laboratory of a university department of medicine, division of pulmonary and critical care medicine. ANIMALS: Ten rabbits after they had received an injection of NNNMU and five control animals. INTERVENTIONS: Controlled mechanical ventilation and bronchoalveolar lavage. MEASUREMENTS AND RESULTS: Measurements of gas exchange (using the multiple inert gas elimination technique), hemodynamics and pulmonary compliance were performed during ventilatory and hemodynamic steady state. A bronchoalveolar lavage (BAL) was taken after sacrificing the animal. BAL samples were processed for cell count and biochemical and functional surfactant analysis. Animals injected with NNNMU developed mild, but significant reduction in PaO2, while maintaining eucapnia during spontaneous air breathing. V/Q distributions and arterial blood gases were similar in all animals when ventilated mechanically with a fixed tidal volume. Compliance of the lung and phospholipid levels in lavage of NNNMU animals was significantly lower than in control animals (CON). Function of surfactant recovered from animals receiving NNNMU was decreased significantly where compared to CON. Thus, NNNMU resulted in a lowered lavage surfactant phospholipid content, impaired surfactant function, decreased compliance and hypoxemia during spontaneous ventilation. However, gas exchange was similar to that of control animals during mechanical ventilation. CONCLUSION: We conclude that NNNMU-induced gas exchange abnormalities present after 4 days are mild and are reversed by fixed volume mechanical ventilation despite marked alteration in surfactant function and lung compliance. These observations further define properties of a lung injury model that is of value in the study of surfactant replacement. PMID- 8708174 TI - The use of inhaled aerosolized prostacyclin (IAP) in the treatment of pulmonary hypertension secondary to pulmonary embolism. AB - OBJECTIVE: To describe the use of inhaled aerosolized prostacyclin (IAP) in a patient with life-threatening pulmonary hypertension secondary to pulmonary embolism and to discuss the possible use of inhaled prostacyclin in the management of pulmonary embolism. DESIGN: Case report. SETTING: Intensive care unit of a university teaching hospital. PATIENTS: One patient with severe pulmonary hypertension secondary to acute-on-chronic pulmonary embolism. INTERVENTIONS: Conventional medical management of massive pulmonary embolism and inhaled aerosolized prostacyclin (IAP). MEASUREMENTS AND RESULTS: Description of clinical course, haemodynamic data and gas exchange data. CONCLUSIONS: We describe a patient with massive pulmonary embolism for whom the addition of IAP to his therapy appeared to result in a transient improvement in pulmonary haemodynamics and gas exchange. PMID- 8708175 TI - Gallbladder abnormalities in medical ICU patients: an ultrasonographic study. AB - Patients in the intensive care unit (ICU) have many risks factors for gallbladder stasis or acute acalculous cholecystitis (ACC), including fasting, total parenteral nutrition, sedation, mechanical ventilation, infection and shock. We have performed a prospective study to estimate the prevalence of ultrasonographic gallbladder abnormalities in 30 consecutive medical ICU patients during the first 2 days of their stay in the ICU. Two patients had previously undergone cholecystectomy and were excluded from the study. Seventeen (61%) of the remaining 28 patients presented with gallbladder abnormalities. Considering three major criteria of ACC, 14 patients (50%) presented with either sludge (25%), wall thickening (22%) or hydrops (11%). However, none of the patients needed a surgical procedure during the study because of gallbladder disease. We conclude that an important proportion of ICU patients presented with gallbladder abnormalities shown by ultrasonography and that this may have implications for establishing a diagnosis of ACC using ultrasonographic criteria. PMID- 8708176 TI - Human recombinant TNF alpha affects rat diaphragm muscle in vitro. AB - OBJECTIVES: To study the effect of human recombinant tumour necrosis factor alpha (TNF alpha) on contracture development and changes in the ionic composition of the external bathing solution of electrically stimulated rat hemidiaphragm preparations in vitro. DESIGN: A triphasic controlled in vitro study using isolated tissue preparations treated in one of four ways. Phase 1 was a control period for all groups; in phase 2 preparations were exposed to TNF, TNF in calcium-free solution, TNF in magnesium-substituted calcium-free solution or no treatment (control); in phase 3 halothane was added to all preparations. The effect of anti-TNF antibody on contracture responses to TNF was studied in separate experiments. SETTING: Laboratories of the Academic Unit of Anaesthesia, University of Leeds, UK. MEASUREMENTS AND RESULTS: Changes in baseline tension and concentrations of potassium and sodium ions in the external bathing medium were measured for each group in each phase. In addition, changes in calcium ion concentration in the external bathing medium were measured for the control and TNF (in calcium-containing solution) groups in each phase. TNF, either in standard Krebs' solutions (p = 0.039) or calcium-free Krebs' solution (p = 0.014), produced an increased incidence of contracture development compared to control, whereas TNF in magnesium-substituted calcium-free Krebs' solution did not. In all three TNF groups there were significant increases in sodium and potassium ion concentrations compared to control, but the changes were significantly fewer in the magnesium-substituted calcium-free group than the two other TNF groups. There was a significant fall in calcium ion concentration in the TNF group compared to control. Anti-TNF antibody reduced the contracture response to halothane in the presence of TNF (p < 0.05). CONCLUSIONS: The results support the hypothesis that TNF acts to increase release of calcium from intracellular stores that are subject to modulation by influx of extracellular calcium or membrane depolarisation. PMID- 8708178 TI - Counting the changes in childbirth: trends and gaps in national statistics. PMID- 8708177 TI - Effects of different triggering systems and external PEEP on trigger capability of the ventilator. AB - OBJECTIVE: The triggering capability of both the pressure and flow triggering systems of the Servo 300 ventilator (Siemens-Elema, Sweden) was compared at various levels of positive end-expiratory pressure (PEEP), airway resistance (R(aw)), inspiratory effort and air leak, using a mechanical lung model. DESIGN: The ventilator was connected to a two bellows-in-series-type lung model with various mechanical properties. Lung compliance and chest wall compliance were 0.03 and 0.121/cmH2O, respectively. R(aw) was 5, 20 and 50 cmH2O/l/s. Respiratory rate was 15 breaths/min. To compare the triggering capability of both systems, the sensitivity of pressure and flow triggered pressure support ventilation (PSV) was adjusted to be equal by observing the triggering time at 0 cmH2O PEEP and 16 cmH2O of pressure support (PS) with no air leak. No auto-PEEP was developed. In the measurement of trigger delay, the PS level ranged from 16 to 22 cmH2O to attain a set tidal volume (V(T)) of 470 ml at a R(aw) of 5, 20 and 50 cmH2O/l/s. The PEEP level was then changed from 0, 5 and 10 cmH2O at a PS level of 17 cmH2O and R(aw) of 5 and 20 cmH2O/l/s, and the trigger delay was determined. The effect of various levels of air leak and inspiratory effort on triggering capability was also evaluated. Inspiratory effort during triggering delay was estimated by measurements of pressure differentials of airway pressure (Paw) and driving pressure in the diaphragm bellows (Pdriv) in both systems. MEASUREMENTS AND RESULTS: There were no significant differences in trigger delay between the two triggering systems at the various PEEP and R(aw) levels. At the matched sensitivity level, air leak decreased trigger delay in both systems, and additional PEEP caused auto-cycling. A low inspiratory drive increased trigger delay in the pressure sensing system, while trigger delay was not affected in the flow sensing system. The Paw and Pdriv differentials were lower in flow triggering than in pressure triggering. CONCLUSIONS: With respect to triggering delay, the triggering capabilities of the pressure and flow sensing systems were comparable with and without PEEP and/or high airway resistance at the same sensitivity level, unless low inspiratory drive and air leak were present. In terms of pressure differentials, the flow triggering system may require less inspiratory effort to trigger the ventilator than that of the pressure triggering system with a comparable triggering time. However, this difference may be extremely small. PMID- 8708179 TI - Neonatal nursing staff time involved with medication-related activities. AB - A work sampling study involving 1920 observations derived from 48 2-hour observation periods is described. Approximately one-tenth of neonatal nurse time was spent on medication-related activities. just over one-third of this time involved medication preparation. It is suggested that the use of a pharmacy medication preparation service may be more cost-effective. PMID- 8708180 TI - Drug administration errors: a study into the factors underlying the occurrence and reporting of drug errors in a district general hospital. AB - Drug administration is one of the highest risk areas of nursing practice and a matter of considerable concern for both managers and practitioners. The aim of this study, carried out in a district general hospital, was to identify any common themes that underlie the occurrence and reporting of drug administration errors. Data were collected from a variety of sources: drug error reports, questionnaires to nurses and nurse managers, and interviews with nurses who had been involved in drug errors. Several areas of particular concern emerged, including: the nurses' confusion regarding the definition of drug errors and the appropriate actions to take when they occurred; their fear of disciplinary action; their loss of clinical confidence; the variation in managerial response; and a possible lack of nurses' mathematical skills. The results from the study demonstrated that it was likely that many drug errors were not reported, for a variety of reasons. It is recommended that all of these issues are addressed as a matter of urgency, for the sake of both patients and nurse practitioners. PMID- 8708181 TI - Assessment and management of children's pain in community hospitals. AB - Registered nurses (n = 72) working in 10 paediatric units in community hospitals in north-eastern Ontario, Canada, participated in a descriptive study investigating how nurses assess and manage pain in children. A four-part questionnaire was used to collect the self-reported data. Twenty-five (36%) of the respondents defined pain as an individual and personal experience and another 25 (36%) respondents defined pain as a more or less localized sensation or discomfort resulting from the stimulation of specialized nerve endings. In response to three different clinical situations, the subjects' mean pain ratings were: 5.72 for an infant; 7.34 for a 3-year-old; and 7.29 for a 12-year-old child. The criterion 'nurses' judgment' was cited as being used frequently in both the assessment and decision making process; however, there was indication that some of the current knowledge in the assessment and management of pain in children was not known or being used. PMID- 8708182 TI - Alternative therapies as control behaviours used by cancer patients. AB - Alternative health care practices are a way in which cancer patients can be in control of their illness situation. This assumption is investigated through control theory. While the sociocultural schemas of North American patients endorse control of illness, the stress of the cancer experience dismisses the advantage of control behaviours. Analysis also suggests that patients find ways to escape from the freedom of being in control of their health situation. This post-analysis of a recent study uncovers major ambiguities concerning the control behaviours of cancer patients. PMID- 8708183 TI - The professional paradigm of qualified psychiatric nurses. AB - The main purpose of this research was to determine the professional paradigm of the qualified psychiatric nurse and the factors influencing the formation of this paradigm. The research was both explorative and descriptive, and both qualitative and quantitative methods were used. The research approach was deductive and based on the theory of paradigm of Tornebohm. The test persons comprised three groups: Swedish-speaking Finns, Swedes and Finns. In each group there were eight students in the final stages of their psychiatric nursing training. A total of 40 questionnaires were distributed to each group. The research revealed four different characteristic types of qualified psychiatric nurses: caring science oriented, partly caring science oriented, general humanist and finally the personality- and experience-oriented. The results also indicate that there is a discrepancy between will and ability within caring. This can partly be interpreted as an expression of the discrepancy between philosophical and ideological impressions and real acts but it may partly indicate a lack of information. Many informants had difficulty naming a theoretical frame of reference for their work and stating aspects of psychiatric caring that would be important to know but on which no information so far exists. Many informants expressed the need for more research and development but did not indicate the subjects. PMID- 8708184 TI - Assessing pain in elderly patients who cannot respond verbally. AB - Many patients cannot respond verbally and there is noted to be an increase in disorientation and confusional states in advanced age. The problem of pain assessment for this group is therefore not only a large problem but potentially a growing one. A pilot study was carried out to validate a chart for the assessment and control of pain using observed pain behaviours. The researchers studied elderly patients with a variety of medical problems on three wards. The study was done by observing pain behaviours, carrying out pain intervention and re observing later to verify the effectiveness or otherwise of the intervention. If this had been successful in moderating the behaviours it could reasonably be inferred that a method of monitoring and comforting verbally unresponsive patients had been found. PMID- 8708185 TI - HIV and homophobia: nurses as advocates. AB - Nurses are frequently encouraged in colleges of nursing and professional journals to consider themselves as ideally placed to act as advocates for vulnerable patients. This paper sets out to challenge this assumption in the light of the stigma and discrimination experienced by people affected by HIV and AIDS. Sexuality and in particular homophobia--the fear and aversion of gay and lesbian behaviour--need to be addressed by the nursing profession. The paper considers concerns about and evidence of discrimination in the light of nurses' claim to be best placed to adopt the role of advocate in representing any patient's health care needs. It is argued that such an assumption is problematical in the face of homophobia and fears around contagion. PMID- 8708186 TI - HIV and AIDS care: consumers' views on needs and services. AB - Increasing numbers of people suffer from HIV infection, in many sectors of society. However, the extent to which health care services meet their needs is open to question. This study aimed to gather consumer views and assess the extent to which care was adequate, through interviews with 85 clients. Their responses were then compared with those of service providers (published in this journal 2 years ago). Overall there was a general expression of appreciation and satisfaction, although in many cases financial and social deprivation was so extreme that it appeared that established services could not compensate. However, the need for better housing and more confidentiality of records was emphasized. The need for greater security from the consequences of prejudice was felt strongly and many distressing incidents were recounted. The major contrast between consumers' and service providers' views focused on whether separate and specialist provision was required. Providers were aware that care would need to be integrated with other generalist services in the future in order to cope with the scale of need, while consumers were keen to retain specialist clinics and wards to protect their confidentiality and sustain the level of expertise they appreciated. Within this sample, however, minority groups wished to have their particular needs recognized, in particular women with children and heterosexual males resented being grouped with the majority group of consumers with HIV/AIDS. PMID- 8708187 TI - Gender-power relations and safer sex negotiation. AB - This paper examines the gender-power relations in heterosexual relationships and discusses the implications for women in attempting to negotiate safer sex. Whilst acknowledging the need for examining both male and female perceptions of sexuality, the paucity of research on men's heterosexual experiences has restricted this paper to the consideration of the woman's standpoint. Reviewing the literature, the complexities of sexual relationships are explored in terms of the power imbalance which arises from the ideologies of femininity and masculinity. The implications for safer sex promotion by practice nurses working in primary health care teams are then discussed within the context of gender divisions of labour and gender role conflict. PMID- 8708188 TI - The health belief model and use of accident and emergency services by the general public. AB - There has been much debate about the use made by the general public of accident and emergency services. A strong element of professional disapproval has been present, as shown by phrases such as 'inappropriate attender'. This paper examines the reasons why people attend accident and emergency and the factors that delay or accelerate attendance, utilizing a framework espoused in the medical sociology literature, i.e. the Health Belief Model. This predicts that individuals carry out a treatment cost-benefit analysis when making decisions about seeking medical assistance. A sample of 200 adult, ambulatory accident and emergency patients was interviewed whilst waiting to see the casualty officer for this study. The data demonstrated that much of the medical, sociological literature concerning patient consultation with doctors is applicable to the accident and emergency situation, in particular the Health Belief Model. A range of factors was shown to make statistically significant differences to the delay times involved in deciding to attend accident and emergency and the time it took to then subsequently attend and register as a patient. These factors also fit the cost-benefit analysis which the Health Benefit Model predicts takes place. Accident and emergency attendance therefore needs to be seen as a logical decision-making process that requires hospitals to provide appropriate services, rather than merely labelling the patients as inappropriate. PMID- 8708189 TI - The multifaceted role of the nurse in the day hospital. AB - This paper presents the initial findings from an action research study designed to explore and improve rehabilitation practice in a geriatric day hospital, The role of nurses in day hospitals and their contribution to rehabilitation form the focus of the discussion, which emanates from the analysis of semi-structured interviews with 13 members of the multi-disciplinary team, and observation of team meetings and patient reviews. Factors which influence the therapeutic activity of nurses are identified and tentative interpretations offered. The findings indicate that the nurse's role in day hospitals is multifaceted and varied and the conclusion is drawn that, far from being marginalized in rehabilitative and therapeutic terms, the nurse's central coordinating role should be recognized as the lynchpin of day hospital activity. PMID- 8708190 TI - The experiences of patients and their partners 1 month after a heart attack. AB - The purpose of this study was to explore any patterns that may be evident in the experiences of 20 patients and their partners 1 month after a first heart attack. An interpretive research approach was used to illuminate the illness experience of patients and its impact on their partners. Semi-structured interviews were conducted with the participants, and qualitative analysis of the data revealed six major categories. These were: expectations about advice and information; feelings about the future; reactions of the partner; playing down the significance of the heart attack; wanting to get back to normal; and the effect on the couple's relationship. These findings are discussed in relation to the theoretical literature and other empirical research. The results of this study may provide pointers to the implications for practice of nurses, particularly on ways to improve support for patients and their partners during early convalescence. PMID- 8708191 TI - Towards an individualized client's care: implication for education. The transcultural approach. AB - The nursing profession is faced with the challenging role of providing individualized client care within the context of the whole person. Individualized care cannot be achieved without considering the factors associated with the personal being, such as culture, beliefs and tradition. As nurses interact with clients from various cultural backgrounds, the need to consider and be aware of their own values is essential to enable them to provide unbiased care. Nurse education plays a major role in developing individuals who are able to examine their own views with regards to cultural biases. The importance of acknowledging other cultures as opposed to the nurses' own is essential in order to ensure that the knowledge learnt will enhance the qualities of caring, competence and professionalism. This is necessary because cultural context is viewed according to how it is perceived by the clients and the nurses themselves. PMID- 8708192 TI - Approaches to community needs assessment: a literature review. AB - In the light of the growing awareness of professionals in the community of the need to undertake health needs assessments of the population, this literature review sets out to explore, delineate and critically analyse the various approaches to community needs assessment, to facilitate a greater understanding of their strengths and weaknesses. The review commences by highlighting its complex nature, and attempting to define what is meant by 'needs assessment' from the differing perspectives of three dominant approaches, namely sociology, epidemiology and health economics. It continues by putting forward an argument for the use of the community health profile, being a multi-focal approach to needs assessment, combining quantitative with qualitative data, and proceeds with a discussion of strengths and weaknesses related to its compilation, in particular factors relating to reliability and validity of data sources. The consumer perspective is also reviewed, as are issues surrounding the ethics of data collection and problems concerning aggregation of the numerous data sources into meaningful policy. Throughout the review, issues are discussed with reference to the current political context in the United Kingdom. Equally important is the community nurse perspective, which is integrated into the arguments where appropriate. PMID- 8708193 TI - Family training: supporting mothers and fathers in the transition to parenthood. AB - This study discusses the views of mothers and fathers on family training and on how family training supports them in the transition to parenthood. The data were collected in 1993, using a questionnaire submitted about 9 weeks after the birth of a child. The same questions were asked of the 189 mothers and the 127 fathers who replied. According to the results both mothers and fathers consider it essential that their views and expectations are given due attention in the planning of family training. Specific issues that were thought to require priority attention in family training were the actual process of childbirth, themes related to parenthood, pain alleviation during labour, and abnormal childbirth. Less importance was attached to the themes of sex, contraception and certain aspects of health education. Both mothers and fathers agreed that they had had ample opportunity to talk with their group leaders and that there had been enough lectures. By contrast, it was widely felt that there was not sufficient opportunity to talk with other group members. It is suggested that efforts be continued to further develop family training schemes so that they offer not only more support to families but also better opportunities for open discussion. PMID- 8708194 TI - Health care rationing: nursing perspectives. AB - Ideas currently postulated around the way health care should be delivered and costs controlled, often referred to as health care rationing, are increasingly coming to dominate the agenda of health care in the 1990s. British nursing has yet to take a noticeably visible role in this debate, despite the fact that it poses a serious dilemma for a profession whose cultural ethos has been shaped by the concepts of universal access and comprehensiveness of care and is wedded to the idea of holism. In the USA debate amongst nurses is further advanced and whilst this discourse may be of limited value to British nursing, owing to a differing historical and cultural attitude to health care, recent changes to the organizational values in the NHS are leading to similar issues arising already faced by American nurses. This paper considers the broad parameters of the debate on health care rationing and examines how these parameters have been reflected within relevant North American and British nursing literature, pointing both to similar and differentiating factors between the two countries. PMID- 8708195 TI - A review of cervical cancer and cervical screening: implications for nursing practice. AB - In this paper we review epidemiological and other research evidence on cervical cancer and cervical screening. We outline recent research evidence implicating sexually transmitted human papillomavirus as one of the causes of cervical cancer, but stress the uncertainty surrounding the causes and natural history of the disease. We go on to discuss evidence on risk factors associated with increased incidence of and mortality from cervical cancer, including age, sexual behaviour, smoking, socioeconomic status and prolonged use of oral contraceptives. Cervical screening has reduced mortality in some countries, and we outline the necessary features of a successful, effective screening programme before going on to describe why screening failed in Britain, at least until the late 1980s. Current screening policy is designed to remedy this, and we discuss its implications, and those of previous research, for nursing practice. PMID- 8708196 TI - A descriptive and exploratory pilot study into school re-entrance for adolescents who have received treatment for cancer. AB - The purpose of this exploratory and descriptive pilot study was to explore the process of school re-entrance for adolescents receiving outpatient treatment for cancers, using grounded theory. All adolescents who had received a minimum of 3 weeks' hospital-based schooling, but had since returned to their own schools, and were booked for an outpatient appointment on a paediatric oncology unit during a specified 2-week period, took part in an audio-tape-recorded, semi-structured interview. The sample consisted of five such adolescents. Data were analysed using the constant comparison method of grounded theory. Certain recognizable trends were noted leading to the development of a core variable to which other categories and their properties related. Central categories identified were 'disruption', 'adaptation' and ,normality', with this latter concept considered to be the core variable. Findings highlighted problem areas which are discussed and issues requiring further detailed investigation. This led to an undertaking of theoretical sampling into hospital-based schooling, as well as suggesting theoretical sampling for future studies. PMID- 8708197 TI - Short or long consultations? Primary care nurses' considerations in making appointments with general practitioners. AB - In Sweden primary care nurses (PCNs) at the health centres (HCs) manage the appointment systems and decide the appointed length of general practice consultation time. This study aims at elucidating experienced PCNs' considerations in booking consultations of different lengths. The grounds for giving long or short appointments were searched for. Qualitative methods were used and 10 PCNs at eight HCs in Umea Health District in northern Sweden were interviewed. The PCNs take factors from two areas into consideration; one area, often immediately apparent, concerns immediate and straightforward issues such as the patient's reported problem, the patient her/himself, the doctor her/himself and the current situation at the HC and in the catchment area. The other area for consideration usually develops through experience over the years and concerns time consumption in a longer perspective. The overall quality of the consultations is then in focus and the length of a separate consultation is of minor interest. The PCNs do not applicate a straightforward relation between length of time and quality of consultations. Experienced PCNs seem to promote accessibility of primary care and their efforts in allocating time in a practical way probably increase the quality of general practice consultation. PMID- 8708198 TI - The community psychiatric nurse in primary care: an economic analysis. AB - Community psychiatric nurses (CPNs) in the United Kingdom are increasingly working in primary health care settings with less serious mental health problems. This paper describes an economic evaluation of their work using a randomized controlled trial in which 231 patients were assigned to continuing general practitioner care or one of two conditions of CPN intervention. This is only the third systematic economic analysis of community mental health nursing in the UK and the first carried out by mental health nurses. Various costs to patients, their families and the health care system were determined. Results showed that patients receiving CPN intervention experienced less absence from work and that this resulted in a net benefit. However, the cost per quality adjusted life year for intervening with this group of patients was probably several times more than for intervening with the seriously mentally ill. Therefore, if one considers both the clinical and economic results of the study, taken together with the recent results of the review of mental health nursing, there seems little justification for CPNs continuing to work in this area. PMID- 8708199 TI - Parallels and contradictions in the theory and practice of action research and nursing. AB - This paper draws on the authors' experiences of nursing in general and of action research projects. Parallels will be drawn between, on the one hand, the relationship between the practice and theory of nursing and, on the other band, the relationship between the practice and theory of action research. Difficulties common to nursing and action research will be identified (e.g. between the implementation of the theory of nursing and of action research). Over- simplification, non-specificity of issues and gaps in theory are frequent but inevitable problems that hinder theoretical applications. Similarly, comparisons will be made between the problems associated with the translation of nursing practice into theory and the difficulties in transposing the experience of research into theory. The complexities of practice, whether nursing or research, and the evolutionary nature of experience inescapably give rise to dilemmas when analysing theory and practice. The concepts of theory and practice when conceived as a dichotomy present a useful overview of issues. We argue that, through a discussion of the core themes of action research, including 'knowledge in action' and 'self and group reflection', consideration should also be given to a symbiotic and complementary analysis of theory and practice. We observe that many contradictions exist within the theory and practice of action research and of nursing, and that these need to be addressed to promote theoretical and practical advances. Action research offers theoretical and practical insights that contribute to the current debate on the reciprocal relationship between theory and practice. PMID- 8708200 TI - An analysis of altruism: a concept of caring. AB - In clinical settings, nurses have observed a conflict within women who are expected to focus on caring for themselves, while their value lies in attending to the needs of others at the sacrifice of self. In this concept analysis of altruism, the term is defined and the critical attributes are identified. Model, borderline, related and contrary cases illustrate what the concept is and what it is not. The meaning of altruism is compared to self-neglect and co-dependence, which are two similar concepts. The antecedents and consequences of altruism are identified to further refine the critical attributes. Implications for research are discussed. This is a first step towards developing nursing theory that will give direction for providing more effective care to women within their value system. PMID- 8708201 TI - The phenomenological movement: implications for nursing research. AB - This paper presents a comparison of the phenomenologies of Edmund Husserl and Martin Heidegger with the aim of highlighting some of the critical distinctions between these two 'schools' of phenomenology concerning the methodological implications of each approach for nursing research. Specifically, the paper examines: the implications of epistemology versus ontology; issues relating to validity; the involvement of the researcher, and aspects relating to interpretation. PMID- 8708203 TI - 'Spirituality as integrative energy': a philosophical analysis as requisite precursor to holistic nursing practice. AB - Spirituality is a universal human phenomenon, yet conceptual confusion, ambiguity and scientific scepticism have prevented adequate investigation into its potential healing effects. A recent resurgence of interest in non-medical sources of healing and holistic medical practices is causing increased speculation regarding the nature of spirituality. The lack of conceptual clarity, and absence of a precise theoretical definition, prevent recognition of spiritual distress and, hence, appropriate nursing intervention. In this paper, current usage and definitions of spirituality are discussed and a philosophical definition of 'spirituality as integrative energy' is proposed. The metaphysical nature of this phenomenon makes it the proper concern of philosophy; consequently, a philosophical analysis of the aforestated proposition was completed using the five predicables of logic theory: species, genus, differentia, logical property and logical accident. Implications of this definition for providing holistic nursing care are also presented. PMID- 8708202 TI - An explanation of the differences between expert and novice performance in the administration of an intramuscular injection of an analgesic agent to a patient in pain. AB - It has been stated that nurse education in the United Kingdom (UK) currently draws upon Schon's concept of the reflective practitioner, and Benner's concept of the expert. Studies have explored the claim that experienced expert practitioners are unable to articulate all they know, and that their theoretical knowledge and clinical skills are of a qualitatively different type from that of novice practitioners. This paper aims to explain the differences between expert and novice performance using the example of administering an intramuscular injection of an analgesic agent to a patient in pain. Theoretical concepts concerning the nature of expertise are used to evaluate novice/expert performance in nursing. Criticisms of Benner's theory are developed in relationship to her methodology and the interpretation and assessment of nursing practice. PMID- 8708204 TI - Health-promoting nursing practice: the demise of the nursing process? AB - Health promotion is gaining widespread recognition throughout the world as the most efficacious practice in achieving health for all. In Canada, the philosophy of health promotion is driving both federal and provincial health initiatives. Such a philosophy is derived from a human science paradigm and is in direct opposition to the natural science paradigm from which the biomedical approach to health care emerged. There now exists a tension between these contrasting paradigms as health care shifts to embrace a health-promotion perspective. The nursing process is based in the natural science paradigm and on a biomedical approach to health care. In order for nurses to embrace health promotion fully, they must move away from the philosophy of the natural sciences and adopt a human science perspective. Such a shift requires a radical transformation in nursing practice as nurses move away from the 'top-down' approach of the nursing process and adopt a 'bottom-up' approach to health-promoting nursing practice. The purpose of this paper is to compare and contrast the nursing process with the principles of health promotion, and to challenge our use of the nursing process in current nursing practice. In particular, a framework for health-promoting nursing practice will be provided. PMID- 8708205 TI - Terms of address: implications for nursing. AB - Nurses are in a position of power over their clients. Power is reflected by status and clients, like all other people, have status relative to their social situation. Terms of address are one of the ways that status and solidarity are demonstrated. They are selected on the basis of the perceived status of the nurse and client, and the solidarity of the relationship. There is a tension between solidarity and status that can cause nurses to select inappropriate terms of address or referring pronouns. The nurse may choose terms of address to increase solidarity in the relationship, for example, to show empathy with the client's apparent vulnerable condition. By increasing solidarity the client's status may seem to be diminished. Clients may have to submit to many invasive, painful, embarrassing or humiliating experiences at the nurse's request. Many of these situations are beyond ordinary acceptable behaviour outside the clinical context. Distress to clients is minimized if they retain as much control as possible and one way keeping a certain amount of control is by maintaining their status. In making a selection of terms of address and pronouns, the nurse must consider the delicate nuances of the options available so that the status of the client is not compromised. This paper discusses the options available in selecting terms of address and referring pronouns, and the sociological implications of those choices. PMID- 8708206 TI - The role of nursing journals in the advancement of professional nursing. AB - From modest developments of 'popular' nursing journals many years ago, there has been a prolific growth in the numbers of nursing journals published each year. This growth is particularly noteworthy in the increase in the numbers of specialist interest and scholarly nursing journals. The different types of journals have also tended to influence each other's contents, as well as the contents of nursing textbooks. Over the years, nurses themselves have emerged as the main contributors to the journals. Increasingly the focus of all the journals has been on nursing practice--and the factors associated with professional practice and the professional education and development of nurses. Whilst it is conceded that it is difficult to demonstrate that nursing journals have made a direct impact on care, some indirect impact can be claimed. This claim is based, among other things, on the contribution of nursing journals to the development of nursing as a profession, their contribution to the individual development of professional nurses, and their contribution to the documentation of the body of nursing knowledge. PMID- 8708207 TI - Disparagement of the nursing process: the new dogma? AB - The nursing process has achieved predominance, if not pre-eminence, in nursing. Concurrent with critical scholarly evaluation of the philosophical foundations of nursing, vigorous critiques of the nursing process have emerged. The critiques are wide-ranging and varied, and question the philosophical commitments, stage of development, components, focus and consequences of using the nursing process. However, the arguments are marred by dualistic dichotomous thinking and a lack of definitional clarity. The critiques fail to discriminate between questions arising from the administrative implementation of the nursing process and questions regarding its theoretical value. Clarification of the definition, description and purpose of the nursing process is essential to rigorous critique and to further development. Given the pervasive use of the nursing process and the need for process theory in nursing, it seems most productive to develop the nursing process more fully and in congruence with contemporary philosophies of nursing. PMID- 8708208 TI - The palliative care quiz for nursing (PCQN): the development of an instrument to measure nurses' knowledge of palliative care. AB - This paper describes the process and outcomes of a study to develop a tool to measure nurses' knowledge of palliative care. The development of the palliative care quiz for nursing (PCQN) entailed the convening of an advisory committee, a wide process of consultation, the development of a conceptual framework, determination of format, generating of items and pre-testing, piloting and further testing of the quiz. The PCQN is a 20-item true, false and 'I don't know' test of knowledge that is easily administered and has utility for assessing knowledge, stimulating discussion and identifying misconceptions about palliative care nursing. PMID- 8708209 TI - A critical analysis of nursing: meeting the spiritual needs of patients. AB - Nursing, over the past 2 decades, has been concerned with the development of theories which will inform practice and help to establish nursing as an emerging discipline. The view that the individual is a biopsychosocial being has occupied a prominent position in the existing catalogue of nursing theories. The literature reveals that, although some existing theories do acknowledge that humans do have the advantage of a spiritual dimension, very little significant work appears to have been done in this area. This paper explores some of the difficulties in arriving at a definition of spirituality and goes on to examine the extent to which nursing theories incorporate the concept of spirituality. Additionally, nurse education does not appear to prepare practitioners adequately to meet the spiritual needs of patients. The paper concludes with a discussion of the potential for meeting the spiritual needs of patients in the context of the changes occurring in the British National Health Service. PMID- 8708210 TI - Clinical nursing research priorities in Australian critical care: a pilot study. AB - The aims of this research project were to identify areas for research in Australian critical care nursing with potential for improvement in patient care, and to advise nursing research policy and priorities with relevance to areas of patients' needs. Research participants represented a national sample of clinical nurse specialists (CNSs) and clinical nurse consultants (CNCs). The Delphi method was used to obtain the most reliable consensus of the specialist nurses, and over 238 research priorities were identified initially. In the final phase of the research nine high priorities were identified. The findings of this study suggest directions for clinical nursing research in critical care. PMID- 8708211 TI - Making sense of action research through the use of a typology. AB - Commenting on earlier work in the Journal of Advanced Nursing on action research, this paper takes up the theme of definition and approach and presents an original action research typology. Three key criteria of action research--'reeducation', 'problem focus', and 'improvement and involvement'--are related to four broad types of action research: 'experimental', 'organizational', 'professionalizing' and 'empowering'. The typology is of general relevance not only to action researchers but to any project concerned to resolve problems and bring about improvements in practice, including evaluation research. Following discussion of the typology, it is used to analyse three different action research projects, including one conducted by the authors in collaboration with a project team. It is shown that each of these projects, which are all concerned in some way to improve standards of nursing care in hospitals, contains elements of different types which may at times be in conflict. The ability to recognize this and to be explicit about the different types may assist action researchers in selecting an appropriate approach and in anticipating problems which may arise during the course of a project's life. PMID- 8708212 TI - British National Health Service trust chief executives on nurse education: corporate instrumentalism and doubts on quasi-market structure. AB - The funding of nurse education through Working Paper 10 arrangements is susceptible to analysis as a 'quasi-market' in which regional purchasers (in England) commission education and training services on behalf of National Health Service (NHS) trusts. In this context, some regions have developed ways of managing the demand side of the market in such a way as to enable NHS trust involvement in purchasing decisions, sometimes through consortium configurations. This paper reports the findings of an empirical study of the views of NHS trust chief executives on nurse education. The investigation was conducted to provide the information base for a consortium development reported earlier in the Journal of Advanced Nursing. The consensus views of chief executives revealed an ideological stance, referred to as corporate instrumentalism. This was shown to be derived from the position of trusts as newly corporate organizations undergoing considerable change in a volatile and competitive environment. Education was seen as potentially capable of contributing significantly to both health service quality and organizational change. Chief executives described an ideal of collaboration in which college providers are highly responsive to their needs, but doubted that the current quasi-market arrangements can in fact deliver this. A comparison of chief executive and senior executive nurse views revealed some significant differences of emphasis, and a mechanism is provisionally suggested by which educational innovation could be suppressed. Finally the education quasi-market was analysed in terms of both its implications for the professional status of nurses and its potential to facilitate NHS reform. PMID- 8708214 TI - Situating theory in practice: student views of theory-practice in Project 2000 nursing programmes. AB - In this paper the views of student nurses taking Project 2000 at a Scottish College of Nursing are reported. A random sample of 19 students were interviewed. The interviews focused on the relationship between the college course and the students' experiences on placement. The majority had difficulty in relating the college course to their practice, except when it came to very specific parts of the programme, for example basic medical procedures. Only a minority could integrate the broader aspects of the course to their practice, largely because there was such a long period of time between studying a topic in college and the related placement. It is tempting to see the results as supporting the situated cognition hypothesis that theory loses meaning out of context, and it is evident, even from this limited study, that the success of Project 2000, with its emphasis on the 'knowledgeable doer', could depend, to a great extent, on the careful phasing of college coursework and placement to enable students to integrate theory and practice more effectively. PMID- 8708213 TI - Virginia Henderson and her timeless writings. AB - This paper provides a reflection on the written works of Miss Virginia Avenel Henderson. Miss Henderson is a nurse, a scientist, an artist and a quintessential human being--all traits which informed her written output. Nursing practice, research and education were all subjects of her extensive chronicle. The four volume Nursing Studies Index is her contribution to nursing research. The Index was sandwiched between two revisions of Principles and Practice of Nursing (5th and 6th eds), the placement of which caused the Index to focus on practice and the Principles (6th ed.) to be based on research. The sixth edition of Principles, written with Gladys Nite and 17 contributors, is considered the most important single professional document written in the twentieth century. The book synthesizes nursing practice, education, theory and research in an age when many nurses are challenged by the seeming incongruity in these essential professional functions. PMID- 8708215 TI - 'Transparency': the missing link in nurse education reform in the United Kingdom. AB - Recent changes to nurse education within the United Kingdom (UK) have proved to be a source of great confusion in many respects, much of which is based on a manifest lack of appreciation of the nature and basis of the changes which are in fact driven by the requirements of European integration. This paper argues that the lack of clarity and publicity, 'transparency', in the process of transposing the requirements of European policy relating to nurse education into UK domestic policy can be blamed for aspects of the lingering confusion within the profession. PMID- 8708216 TI - Role conception and role discrepancy: a comparison between hospital-based and degree nursing students in Hong Kong. AB - This paper reports on a cross-sectional survey that aimed to compare the role conception types and role discrepancy between 140 certificate and 81 degree nursing students in Hong Kong. Non-probability convenience sampling was used and the participants selected from the hospital-based training were first, third and fourth study block students. Second-, third- and fourth-year students from the degree programme were selected for comparison. The role conception scale developed by Pieta was modified and adopted to measure the role conception and discrepancy. ANOVA test was performed to examine differences across and within the two groups. The results showed that the degree students had a significantly higher ideal but lower actual professional role conception than the certificate students. An interaction effect between the groups (certificate--degree) and grades (stages of education) was detected in the ideal professional role, indicating a marked drop of the score for the fourth-year degree students. For the discrepancy role conception score, the degree students had a significantly higher discrepancy score in all the three role conception types compared with their certificate counterparts. In summary, the impact of the educational process on the development of role conception was demonstrated. Compared to the certificate students, the ward practice seemed to exert a far greater impact on degree students' role conception. PMID- 8708217 TI - Conflict, enactment, empowerment: conditions of independent therapeutic nursing intervention. AB - The purpose of this qualitative study was to describe the use of independent therapeutic nursing interventions by registered nurses in a variety of health care delivery agencies. The process used to integrate nursing interventions into the daily routines of agencies that did not formally require their use, the conditions in which they were implemented, and the consequences of their use, were examined. Data collection methods include observations and interviews of 36 participant nurses, employed in five health care agencies, for 1 year. The organizational culture of each environment was observed and its impact on the enactment of independent therapeutic nursing interventions was evaluated. Data analysis, done by using the constant comparative method, generated a proposal of a substantive theory that integrates conditions and consequences of therapeutic nursing intervention. Intra-role conflict, the product of incompatible expectations of the professional conception of nursing and task oriented/medically controlled work environments, served as the motivating force behind strategies of enactment utilized to overcome barriers to implement therapeutic nursing interventions. As a consequence of using therapeutic nursing interventions, participants perceived empowerment in relation to clients, peers and other health care professionals. PMID- 8708218 TI - Postmodernism, health and illness. AB - This paper examines the value of drawing on ideas from poststructuralism and postmodernism in an attempt to understand the relationship between health, culture and society. Medical and professional discourses have come under increasing criticism for being uncaring, stigmatizing and disempowering. This paper supports a postmodernist approach which allows analysis of the fabrication of 'health', 'illness' and 'patient' subjectivity and the effect of the inscription on the body. It is suggested that health care professionals need to be more reflexive about their own knowledge claims and to resist the discursive practices which disempower and reduce choice. PMID- 8708219 TI - Second nursing symposium: 'Routes from Research to Reality' held at University College of St Martin, Lancaster, England, 7 April 1995. PMID- 8708220 TI - Is the professional code still the cornerstone of clinical nursing practice? AB - Although codes of practice for those concerned with the health care of others have always been inherent in the structure of societies, they have been institutionalized within the nursing discipline since the end of the last century. Up until the early 1970s they promulgated subservience to the medical discipline. As a result of the processes of emancipation and professionalization, the philosophy of the nurse has come to contain concepts of autonomy, accountability and patient-advocacy, based on a personal and individualized care system. Research in recent years has shown that nurses are making morally sound and ethically acceptable choices based on their own decision-making abilities, whilst having little or no active knowledge of the existing professional codes. Based on the literature, the author discusses ethical codes in relation to their perception by nurses in the clinical situation. The influence of the code in the areas of moral decision making, administration and management, and education are likewise discussed and the conclusion is reached that codes remain the cornerstone of nursing practice. PMID- 8708221 TI - The use of different research methodologies to evaluate the effectiveness of programmes to improve the care of patients in postoperative pain. AB - Much research has supported the conclusion of the report from The Royal College of Surgeons and College of Anaesthetists (1990), London, England, that the relief of postoperative pain is in many cases unsatisfactory and calls for more research on the effectiveness of educational programmes. A prime aim of nursing research is to influence and to improve practice. In relation to postoperative pain Sofaer's (1985) study was an example of the use of a quasi-experimental approach to improve postoperative pain relief through nurse education. Sofaer suggests that although positive effects were demonstrated they may not have been sustained in the long term. Further, Sofaer suggests that action research may have produced a more sustainable change. This paper critically discusses the use of these two research methods in relation to this problem. PMID- 8708222 TI - Living with ulcerative colitis: experiences of adolescents and young adults. AB - The problems associated with ulcerative colitis and its treatment have effects on adolescents and young adults dissimilar from as well as more profound than those on older individuals. Adolescents are confronted with problems such as biological, psychological and social changes as well as role changes related to peers and family. This inductive study aimed to describe the adolescents' experiences of living with ulcerative colitis. A total of 28 subjects were asked about their experiences both at the present time and at the time their first symptoms appeared. Verbatim transcribed thematized interviews were analysed according to a method influenced by the constant comparative method for grounded theory. Eight categories were grounded in the data, forming a model which describes the process from onset of disease to present time. The main variable identified was: reduced living space, a strategy to manage the new situation. Dependent on the reactions received from significant others, the outcome for the adolescents hovered between feelings of self-confidence and lack of self confidence. If the adolescents experienced support, the living space was expanded again. The results might be of great value when caring for and assisting young persons with a chronic disease in general, and in particular when taking care of adolescents with a recently diagnosed inflammatory bowel disease. PMID- 8708223 TI - An examination of nurses' practices when performing aseptic technique for wound dressings. AB - The purpose of this study was to establish if nurses' actions when carrying out 'aseptic technique' using the 'gloves technique' are simple and based on up-to date knowledge and do not incur unnecessary wastage. A sample of convenience was used involving 21 trained nurses. Observation and formal interviews were used to collect quantitative and qualitative data. Results showed that not all nurses in the sample applied a 'simple aseptic technique'. The rationale for the practice of aseptic technique was not always research based, though other aspects of wound management were derived from research findings. The study highlighted other areas of aseptic technique which require investigations. PMID- 8708224 TI - The effect of body positioning upon maximal oxygenation of patients with unilateral lung pathology. AB - A quasi-experimental, repeated-measures cross-over design study on the effect of body position on oxygenation (SaO2) blood pressure, respiration and pulse in patients with unilateral lung pathology was conducted. Previous research strongly suggests that positioning with the healthy (unaffected) lung in the dependent lateral (down) position is related to improved oxygenation, but knowledge about whether this effect is maintained over time is lacking. The purpose of this investigation was to determine: (1) Is positioning with the unaffected lung in the dependent lateral position related to increased arterial blood saturation levels and decreased blood pressure, pulse and respiration? (2) What is the relationship between the dependent variables--oxygenation saturation levels, blood pressure, pulse and respiration--and the independent variables--body position and time in the position? Thirty-nine patients with unilateral lung pathology were positioned on their sides with the unaffected lung down, on their sides with the affected lung down, and also in semi-Fowler's position. Arterial (SaO2) blood saturation and vital signs were measured at baseline 0, 15 and 30 minutes. There was no statistically significant relationship between oxygenation level or systolic blood pressure. Diastolic blood pressure, respiration and pulse did vary significantly with position. PMID- 8708225 TI - A comparison of the ability of foam swabs and toothbrushes to remove dental plaque: implications for nursing practice. AB - This study aimed to assess the impact which the use of a toothbrush and the use of foam swabs had on the removal of dental plaque over a 6-day period. Three experiments were completed and duplicated using the author's mouth and one of these experiments was completed a third time on a volunteer's mouth. A plaque scoring system which quantified the amount of plaque on teeth in areas adjacent to periodontal tissue, and therefore capable of initiating inflammation, was used after plaque had been disclosed. Plaque at the gum/tooth margin (gingival crevice plaque) and plaque between teeth (approximal plaque) was measured. At the end of each 6-day period, which commenced with all tooth surfaces clean, the ability of the toothbrush to remove plaque was consistently better than that of swabs, and usually achieved complete visible plaque removal from all sites. In contrast, plaque remained in all sites which had been cleaned using foam swabs (after using a 'swabbing' or 'scrubbing' technique). However, it was possible to remove plaque from a number of gingival crevice sites with a swab when a varied 'any technique' was used on the visible disclosed plaque. The plaque in all approximal sites still remained after this technique. An experiment to measure the effect of using foam swabs on plaque which had been allowed to accumulate over a 6 day period produced similar results. The results from this study suggest that the success of a toothbrush in removing plaque is affected by user technique (total visible plaque removal was not achievable), and that foam swabs are not able to remove plaque from some 'sheltered' areas of teeth (total visible plaque removal was not achievable). The implications of these findings to nursing practice are discussed. PMID- 8708226 TI - Social support and its impact on mothers' experiences of childbirth. AB - The purpose of this study was to look at the networks of social support available for mothers during pregnancy and childbirth. Another concern was to describe mothers' experiences of labour and the connections of social support with those experiences. The data were collected through the use of two questionnaires submitted to 200 mothers recruited from Tampere University Hospital. Both instruments--Norbeck's Social Support Questionnaire and a scale designed by the author--were based on Kahn's (1979) theory of social support. The mothers reported an average of seven support persons each. Most typically, these were the mother's spouse or partner and close friends and relatives. The network of social support is a major source of emotional support, aid and affirmation for the mothers. During labour the main source of emotional support for mothers was the midwife. For the vast majority (85%) of the mothers, childbirth was a positive experience. A significant association was found between the emotional support provided by midwives and mothers' positive experiences of childbirth. PMID- 8708227 TI - Social support: some pragmatic implications for health care professionals. AB - The role of social support in promoting recovery from chronic illness has been the focus of a debate within the nursing and social science research communities. This paper reviews the literature on this important issue and discusses the implications for patient management. In providing holistic patient care, health care professionals need to reflect on the impact of this research for their clinical practice. PMID- 8708228 TI - Employed Mexican women as mothers and partners: valued, empowered and overloaded. AB - This study was designed to explore the daily lived experiences of a group of employed, low-income Mexican women in their maternal and spousal roles. The participants were 41 auxiliary nurses recruited from two large urban hospitals in Mexico. Data were collected through the Women's Roles Interview Protocol (WRIP), which solicited the participants' perceptions of the satisfaction and stresses they experienced in their roles as mothers and spouses, and their descriptions of the coping strategies and the resources they used to deal with stressful life experiences related to these roles. Data analysis consisted of a qualitative thematic analysis of the narrative responses to open-ended questions in the WRIP. Satisfying aspects of the maternal and spousal roles, as identified by the participants, included giving to and receiving from their children and being valued and supported by their partners. Spousal approval of their work was also satisfying. These employed mothers, however, experienced many stressful aspects of functioning in multiple roles, including lack of resources, being absent from their children, self-doubt about their maternal role functioning, role overload and spousal absences. The women coped by juggling priorities and utilizing family resources. From the data analysis, the investigators developed a conceptual framework for understanding these women's experiences with parenting and marriage. The centrality of the family, a sense of value and empowerment as women in maternal and spousal roles, and the reality of role overload are discussed within the Mexican culture context of machismo, its female counterpart hembrismo, and family life. Implications for women's health are framed within a context of family and work. PMID- 8708229 TI - Teaching family nursing: strategies and experiences. AB - The traditional hospital-based approach to Australian nurse education curricula was primarily based on the medical model and directed towards the preparation of nurses who were able to give care to individual clients. The major focus was on the needs of the individual. A notable absence in curricula was any consideration of the role or importance of families to individual and family health. This was despite the continuing involvement that nurses have in their practice with the families of their clients. This paper describes the experiences of introducing a family nursing subject in an undergraduate, preregistration nursing programme which focuses on the family as a unit of care. Educational strategies, clinical experiences, and evaluation of the unit of study are discussed. PMID- 8708230 TI - A qualitative evaluation of the post accident notification system to health visitors. AB - Childhood accidental injuries are a major public health problem, whether measured in terms of morbidity or mortality, and accident prevention is subsequently a priority issue. The Accident and Emergency Notification System to health visitors provides them with vital information and statistics with which to plan accident prevention work, both on an individual basis or in group work. This small study aimed to look at this system from the health visitor's perspective and explore the action taken. The grounded theory approach was used to uncover the processes used by the health visitors. Data were collected using unstructured interviews and by writing notes. These were transcribed on the same day as recorded. Nine categories were identified and an analysis of the processes involved when receiving the notification slip is presented. The categories included 'knowing the family', 'age and development', 'pressure of work' and 'gut feeling/extra sensory perception'. This study is a small contribution to our understanding of the processes involved in everyday health visiting practice. PMID- 8708231 TI - The merger of United Kingdom colleges of nursing with university departments of nursing: prospects, problems and promises. PMID- 8708232 TI - Functional health status and instrumental activities of daily living performance in noninstitutionalized elderly people. AB - The purpose of this descriptive correlational study was to examine the relationship between functional health status and the performance of instrumental activities of daily living (IADL) in a group of noninstitutionalized elderly people. A convenience sample of 47 subjects who were 70 years of age or older participated by completing three questionnaires: the Health Status Questionnaire, the Multidimensional Functional Assessment IADL Scale and a demographic questionnaire. Lawton's (1985) theoretical model of person-environment interaction provided the conceptual framework for the study. Three functional status variables reached statistical significance (0.001) with IADL performance: social functioning (r = -0.6045); health perception (r = -0.6984); and physical functioning (r = -0.6635) suggesting that these are important indicators of overall health status, and that a decline in each could contribute to increased IADL dependency. The IADLs of housekeeping, shopping, and transportation correlated significantly (0.001) with functional health status, implying that, as health declines in elderly people, they may require more assistance with IADLs, in order to remain more independent in the community. The results have implications for nursing administrators and nursing educators with regard to developing health promotion strategies related to independence of elderly people, community health and social policy. PMID- 8708233 TI - Undernutrition and elderly hospital patients: a review. AB - Numerous surveys have revealed that many hospital patients are undernourished. Elderly patients are particularly vulnerable to the adverse consequences of undernourishment. Slow acquisition and application of available knowledge has been blamed as a principle reason for lack of progress in tackling the problem of 'hospital malnutrition'. This paper provides a resume of the knowledge accumulated from research with the purpose of promoting active and informed nursing involvement in the detection and management of undernutrition in hospital wards. PMID- 8708234 TI - Pain and elderly patients: a survey of nurses' knowledge and experiences. AB - Two hundred and eight qualified hospital nurses each completed a short questionnaire about pain and pain control in patients aged 70 years and over. Their awareness of the prevalence of chronic pain and its negative consequences was good, as was their knowledge of methods of assessing pain. Many, however, harboured misconceptions about the pharmacological treatment of pain and exaggerated the risk of opioid-induced respiratory depression. A wide range of complementary methods of pain control had been used by many of the nurses, and considerable interest was expressed in developing these skills further. Nurses from acute surgical areas tended to have a better knowledge of issues related to pain in elderly people than those from care of the elderly wards. PMID- 8708235 TI - Relationship between elderly carers and the older adult with learning disabilities: an overview of the literature. AB - This paper presents an overview of the literature concerning the changing relationship of the elderly carer and the older adult with learning difficulties. It identifies the need to further explore the relationship between informal carers as they age and the older person with learning difficulties. The paper suggests that life changes and choices by older carers will add strain to the relationships between them and those for whom they care. These episodes will be exacerbated by the ageing of the person with learning difficulties. The lack of understanding of this changing relationship makes the adequate provision of professional care unlikely. There is a need to address how health and social services can work with both the carers and the clients to manage the process of ageing and maintain the support needed by those at risk--older adults with learning difficulties and the people who care for them. PMID- 8708236 TI - Adjustment and social behaviour in older adults with chronic obstructive pulmonary disease: the family's perspective. AB - This study employed secondary data analysis to explore family perceptions of adjustment and social behaviour in older adults (n = 51) with chronic obstructive pulmonary disease (COPD) and their relationship to published norms and patient self-report. According to the Katz Adjustment Scale for Relatives, these COPD patients had significantly higher levels of belligerence, negativism, helplessness, withdrawal, psychopathology, nervousness and confusion than reports from relatives of older adults from the general population. No differences were found in performance, expectation or dissatisfaction with socially expected activities, or performance of free-time activities. However, family members of COPD patients were significantly more dissatisfied with their relative's free time activities. Although family perceptions of socially expected activities corresponded to patient descriptions of general and physical functioning (Sickness Impact Profile), patient perceptions of psychosocial functioning were independent of the family's. The results supported the tenet that older adults with COPD have difficulties with adjustment that may adversely affect social relationships, but were not consistent with the belief that the performance of socially expected or free-time activities is more impaired than in others of this age group. The data also suggested there may be some perceptual discrepancy between family and patient views of social behaviour. PMID- 8708237 TI - A presentation of a conceptual framework and its use in the definition of nursing development within a number of nursing development units. AB - This paper provides a conceptual framework upon which action plans can be built to enable registered nurses to reflect upon their work and practice activities. This conceptual framework has been used by the author in his work with various nursing development units within Yorkshire, England, in his role as their academic nurse advisor. The framework has been useful in helping nurses not only to reflect upon current work and practice activities but also as a guideline for future development. The paper provides examples of such nursing development activities and includes a personal view of how nursing development units can act as catalysts for change. PMID- 8708238 TI - Nursing development units: an opportunity for evaluation. AB - The nursing development unit movement has recently generated a lot of financial and professional interest. This is demonstrated by the large financial investment by the Department of Health, England, and the growing number of papers in the nursing literature. The benefits of these units, however, are not as well documented. This paper seeks to contribute to the debate by: reviewing some of this literature and putting the movement in a historical perspective; discussing the purposes of the units; critically examining the characteristics of such units; evaluating their effectiveness; and suggesting a way forward for the future. PMID- 8708239 TI - The experience of reassurance: patients' perspectives. AB - This study sets out to examine reassurance from the perspective of patients from a medical and a surgical setting in a local general hospital unit. The rationale for studying reassurance is discussed and it is indicated that the phenomenon of reassurance as a caring construct is under-researched. The aims of the study are listed and phenomenology is used as the research method to answer the research question posed. The participants were asked to describe their experiences of being reassured by the nurses. Unstructured interviews were the main method of data collection and the sample consisted of eight patients of mixed gender. Data were analysed using Colaizzi's procedural steps and these are described. The process of research is described and ethical considerations discussed. The results are described using raw data from the subjects. The emerging themes include perception of the environment, 'being with', receiving information and knowledge of facts, interpersonal skills, 'being there', communication skills, trusting relationship, being cared for, assertion of optimism, and humanistic traits of the nurses. In the conclusion, the limitations of the study are briefly mentioned. The relevance of the findings to nursing practice are addressed and tentative recommendations for further research are made. PMID- 8708240 TI - Training hospice nurses to elicit patient concerns. AB - Patient assessment underpins every aspect of nursing care. However, there is much evidence to suggest that many nurses lack the skills necessary to communicate effectively with their patients, and so assess their individual problems and concerns. Communication studies to date have been descriptive, or have concentrated on acquisition of skills without addressing the impact this has on patient care. This paper reviews a study of 44 hospice nurses who were taught assessment skills. It discusses the impact of training not only on their skill level, but also on their ability to elicit their patients' concerns. It concludes that simple skills training is insufficient to change clinical behavior, and discusses other factors which should be addressed in future training programmes. PMID- 8708241 TI - Cultural perspectives of grief: an expanded concept analysis. AB - The concept of human grief has been of interest to scholars and practitioners for many years. However, there are many aspects of grief about which little is known. One of these is the role that cultural heritage plays in influencing the individual, intrapersonal experience of grief. Through the use of six focus groups, each consisting of persons from a specific cultural background, the investigator explored the concept of grief. The data were thematically analysed and the findings then compared to the findings from a previous concept analysis of grief drawn from professional literature. The findings indicated that individuals from a variety of cultural backgrounds whose knowledge of grief was derived primarily from their personal experiences defined grief in much the same way as had the authors of the professional literature. Most importantly, the findings also clearly indicated that although cultural differences are perceived to exist in mourning rituals, traditions and behavioural expressions of grief, there are no particular differences in the individual, intrapersonal experience of grief that can be attributed to cultural heritage or ethnicity alone. Implications for future research and considerations for practising nurses as they pertain to the highly individual and pervasive nature of grief are presented. PMID- 8708242 TI - Why I want to be a psychiatric nurse: constructing an identity through contrasts with general nursing. AB - The justification of psychiatric specialization among undergraduate nursing students is examined. The data highlight how students are able to produce a positive view of their initial entry into nursing and then later create a positive and distinctive specialist occupational identity through negative comparisons with general nursing. Their experience in both psychiatric and general settings is drawn upon to add credibility to their claims. Contrasts based on the work schedules, patient focus, professional autonomy, opportunity for professional development, and required skills are considered. PMID- 8708243 TI - 'Coffee & condoms': the implementation of a sexual health programme in acute psychiatry in an inner city area. AB - A sexual health programme was set up specifically to target mentally ill individuals, providing a distinct group of interventions designed to assist this specialist cohort in acquiring knowledge, skills, and attitudes that will directly contribute to their mental health. The programme consisted of gender specific and mixed groups, a drop-in service, condom provision and a full-time programme coordinator. Areas covered were safer-sex, knowledge of HIV/AIDS and other sexually transmitted diseases, assertiveness and practical skills of condom use. Mental health nurses possessed the skills to offer sexual health advice to mentally ill individuals. Mentally ill individuals were enthusiastic to attend groups, and were able to participate in the programme. PMID- 8708244 TI - An analysis of the concept of empowerment. AB - This paper is an analysis of the concept of empowerment and its use in nursing practice, education, research and health promotion. The paper adopts an eclectic approach to concept analysis, incorporating the methods advocated by Walker & Avant and Rodgers. The concept is analysed and a theoretical definition given. Defining attributes, related concepts, antecedents and consequences of empowerment are proposed and a model case presented. The analysis demonstrates that empowerment is: a helping process; a partnership valuing self and others; mutual decision making; and freedom to make choices and accept responsibility. Implications for practice conclude the paper. PMID- 8708245 TI - How nurses in a stroke rehabilitation unit attempt to meet the psychological needs of patients who become depressed following a stroke. AB - Fourteen qualified nurses working in a stroke rehabilitation unit were interviewed in order to ascertain their understanding of post-stroke depression, what they do to help a patient who becomes depressed and what they feel unable to do. They were also asked what would improve the situation. The nurses were able to describe the characteristics of depression following a stroke and recognize its effect on the subject's rehabilitation. They were able to identify patients who were becoming depressed and tried to help them to the best of their abilities. They felt constrained, however; mainly by lack of time, limited skills and lack of appropriate training. They themselves wanted to be able to meet the needs of depressed patients and thought that this could be achieved through better staff education and access to other health professionals experienced in providing psychological care. It is recommended that nurses have access to expert personnel both as a source of referral for patients and as a resource for support and guidance to staff. In addition, an education programme combining theoretical and practical aspects of psychological care would be useful. PMID- 8708247 TI - An evaluation of a Canadian peer-driven injury prevention programme for high-risk adolescents. AB - The mortality and morbidity resulting from serious trauma in adolescence, particularly head and spinal cord injury, constitutes a health problem of major proportions. Although many community-based prevention programmes have been reported in this last decade, few of these describe an evaluation component. In this study, a school-based prevention programme was developed by a peer group and presented by them to high-risk adolescents. The study aimed to test the efficacy of this intervention compared to the delivery of a prevention presentation to a similar group by a health care professional and compared to a control group. Measures of health locus of control, self-efficacy and behavioural intent were supplemented by open-ended items related to risk-taking behaviour change. At post test and at 4-month follow-up, there was little evidence in the quantitative measures to support the effectiveness of the intervention for reducing injury risk factors. More encouraging findings were seen in the qualitative data. Explanations for why the intervention did not result in the expected outcomes are offered. PMID- 8708246 TI - Australian registered nurse medical device education: a comparison of simple vs. complex devices. AB - As the primary users of medical devices in direct patient care, registered nurses must be competent both with simple and complex devices because any device improperly used can have serious consequences. Results of a cross-sectional survey of registered nurses (n = 238) working at a large South Australian tertiary care centre on units with high usage both of indwelling urethral (Foley) catheters and intravenous infusion pumps revealed that participants were significantly more likely to have learned to use the indwelling urethral (Foley) catheter in nursing school than they were to have learned how to use an intravenous infusion pump. Participants differed significantly on most (13 of 17) items about what they learned, with nurses being more likely to learn various factors about the indwelling urethral catheter than the infusion pump. Registered nurses were more likely to feel stressed when using an intravenous infusion pump than when using an indwelling urethral catheter, but were more likely to have used an indwelling urethral catheter that caused a patient harm than an intravenous infusion pump that caused a patient harm. The most frequently identified reasons for incidents of patient harm resulting from use of an intravenous infusion pump were user error and inadequate device education. The most frequently identified reasons for incidents of patient harm resulting from use of an indwelling urethral catheter were the patient's condition, particularly confusion, user error, equipment malfunction, and inadequate device education. PMID- 8708248 TI - The use of qualitative methods in the study of parents' experiences of care on a children's surgical ward. AB - User satisfaction with the care of hospitalized children is examined in this paper. It is argued that parents are proxy consumers but that their status is conditional on the way in which they exercise choices. Parents' involvement in the care of their children means that the term 'user' is more appropriate than 'consumer'. Methodological problems in the assessment of user satisfaction are discussed. A study of the experiences of parents of 24 children discharged from a surgical ward of a children's hospital is described and the implications for the assessment of user satisfaction reviewed. The study demonstrates that qualitative methods have value in the assessment of user satisfaction with care. A particular strength of qualitative methods of inquiry is that limitations of users' choice in care can be assessed. PMID- 8708249 TI - Understanding the needs of relatives of patients within a special hospital for mentally disordered offenders: a basis for improved services. AB - This paper outlines one component of a major research study, which will form the basis for a new service for relatives of schizophrenia sufferers within a special hospital. The aim of this initial component has been twofold: firstly to identify and understand the needs of relatives in terms of their present knowledge and understanding of schizophrenia; and secondly to establish what stress the relatives are presently experiencing in maintaining contact with patients and whether this stress may have an impact on the management of schizophrenia at the hospital. A purposive sample of 17 relatives were interviewed using two semi structured interview protocols. All interviews were audio-taped and later transcribed for analysis. The data have provided an illuminating insight into the thoughts and feelings of relatives and indicate many ways in which a service for relatives could be tailored. It is apparent that many relatives are currently not being supported, involved or allowed to contribute to the care of their family member at Ashworth Hospital. This present situation is stressful for relatives and is likely to have a detrimental effect on the course of schizophrenia in patients at Ashworth Hospital. PMID- 8708250 TI - The factors influencing organ donation: a review of the research. AB - Transplantation of organs has become the treatment of choice for many conditions, yet there remains a worldwide shortage of donor organs. This paper reviews the factors which influence organ procurement. It concentrates on the public's and the professional's attitudes and examines the potential role research could play in overcoming these difficulties and thus increasing donor referrals. PMID- 8708251 TI - Problem-based learning in an undergraduate nursing programme: a case study. AB - Over the past 15 years, the 'theory-practice gap' has been a recurrent theme in the nurse education literature. Numerous explanations are put forward for its existence, along with many suggestions as to how the apparent divide can be bridged. A key component of nursing practice is problem-solving but not all nurses are competent at finding suitable solutions to the problems they face in practice settings. Therefore ways of developing learner proficiency in problem solving is crucial and should occupy a substantial part of teacher activity. One recent educational method for promoting problem-solving skills is problem-based learning [PBL] using the hypothetico-deductive technique. This approach encourages students to work through problem situations, generating hypotheses and testing these against the relevant literature and personal experience. The process itself is seen as the essential element in developing problem-solving skills, so that when the students are qualified they can apply the same methods to patient care. This paper explores the use of the PBL approach with a group of 11 fourth-year undergraduate students. The method employed was a case study design using observation as the main data collection technique. Subsequent analysis focused on: (a) practice; (b) teaching method; (c) knowledge attainment; and (d) the role of the teacher. PMID- 8708252 TI - Exploring the process of data acquisition: methodological challenges encountered and strategies employed. AB - Data acquisition marks the first phase of the problem-solving process. Little research exists, however, on data acquisition amongst nurses despite its centrality in the problem-solving process. In part, this is a reflection of the inherent challenges and difficulties posed by this area of research. This paper discusses methodological issues raised during a recent study exploring the process of data acquisition amongst student nurses. The research forms part of a larger comparative study of the outcomes of preregistration nurse education programmes, funded by the English National Board for Nursing, Midwifery and Health Visiting. PMID- 8708253 TI - A study of nurses' attitudes towards research: a factor analytic approach. AB - Despite considerable top-down pressure on nurses to undertake research, the available evidence suggests that relatively few nursing studies are submitted for publication. Given the need to increase the degree to which research informs practice, this shortfall in output must of necessity constitute a cause for concern. Therefore, it seems timely that a full-scale investigation into the possible reasons for non-submission is conducted. This study, therefore, in an attempt to meet this aim, used factor analysis on the scores from an attitude-to research scale completed by 230 nurses. The results suggested that five coherent factors underpinned the sample's general attitudes to research. These were labelled 'nurses' subjective barriers to research', 'organizational/structural barriers to research', 'doctors' reactions to nursing research', 'health care professionals' reactions to research' and 'impact of nursing research'. The variables clustering on two of these factors suggested a predictive relationship with two relevant outcome behaviours relating to research. When this was tested, the predictions were supported, indicating that these factors could be used as a quick and simple screening tool to highlight individuals or groups of nurses who might benefit maximally from specific attitude change programmes. In this way, it might be possible to increase nursing research activities and output. PMID- 8708254 TI - The similarities and differences between nurses with different career choice profiles: findings of an interview survey. AB - This paper explores differences and similarities between participants who had selected nursing as an occupational choice from childhood and those for whom nursing was not a childhood occupational choice. The implications of the findings are discussed. PMID- 8708255 TI - Gender differences in career progress in nursing: towards a non-essentialist structural theory. AB - This paper briefly reviews the literature on gender differences in career progress in nursing. It argues that current explanations for these differences are essentialist in nature. Aspects of the literature on the labour market and neo-Weberian closure theories are critically reviewed and a synthesis of these two domains is proposed. The paper argues that this synthesis can provide a non essentialist explanatory framework for gender differences in career progress and provides a foundation for ongoing empirical work. The exercise points to the importance of geographical mobility and its use as an exclusionary criterion in career progress. In the context of the nursing labour market this process may serve to maintain structural gender bias in professional nursing. PMID- 8708256 TI - The changing role and work of British nurse tutors: a study within two demonstration Project 2000 districts. AB - This study was designed to describe the main changes in the role and work of nurse tutors brought about by Project 2000 initiatives, with a view to exploring what the work of nurse tutors would be in the future. It was centred upon two demonstration Project 2000 districts, chosen to provide contrasts. Data were collected over a 15-month period from a range of stake-holders, but with the priority on nurse tutors themselves, using semi-structured individual and group interviews. The results showed that, in future, the role and work of nurse tutors would adapt to reflect the integration into a higher educational environment. The main changes expected include: alternative approaches to teaching and learning, for example student-centred teaching and open modes of study; greater subject specialization; more teaching across 'schools'; larger class involvement; more effective educational organization and management; closer liaison with the service provider units; and more effective time management. The study found that nurse tutors would be required to diversify their work through new initiatives, to undertake consultancy and research and to compete more for resources- developments which were not universally welcomed. PMID- 8708257 TI - Clinical evaluation--difficulties experienced by sessional clinical teachers of nursing: a qualitative study. AB - Evaluation of nursing students in the clinical field requires the clinical teacher to make judgements regarding student progress in a number of areas. In this study concepts of role theory, oppressed group behaviour and the ethics of caring emerged and were used as conceptual frameworks to interpret the data relating to the evaluation of undergraduate students. The number of experienced faculty available for clinical teaching and evaluation has become inadequate and a large number of casual or sessional clinical teachers are employed to teach students in the clinical field. Despite the well documented problems associated with clinical teaching and the use of inexperienced clinical teachers, sessional clinical teachers are nevertheless expected to evaluate student success in meeting the clinical requirements of the nursing course, often resulting in disparate decisions for students. A phenomenological study was carried out using unstructured interviews and written clinical scenarios, to explore the evaluation process from the perspective of the sessional clinical teachers. Research findings indicate that although the sessional clinical teachers were skilled at identifying student problems, they were reluctant to make difficult evaluation decisions, due to low self-esteem, role conflict and their ethic of caring. It seems that gender socialization, patriarchal dominance and apprenticeship training had effected their confidence in their own decision making. The implications of such findings are of concern for the ongoing credibility and integrity of nursing courses, as clinical teachers have an influence on the nursing profession through the preparation of its practitioners. PMID- 8708258 TI - Psychiatric nursing and Project 2000. PMID- 8708259 TI - Critique of collaboration. PMID- 8708260 TI - The role of computer-based resources in health promotion and disease prevention: implications for college health. AB - The development of the Internet-based World Wide Web communication platform has opened a new avenue for extending the program effectiveness of college health centers. California College Health 2000, a San Diego State University-based HIV prevention project, is collaborating with the university's student health center to develop a computer-based resource to provide hypertext links to specific health information helpful to health promotion planners and consumers. The project's virtual resource centers are organized around health topics and seek to encourage the dissemination of health information to the broadest possible constituency of college health professionals and are designed to provide for continuous feedback and to evolve over time. Implications for the future of college health, based on the creation and pilot use of the resource centers, are discussed, and recommendations are made regarding collaboration, leadership, and continuing innovations in the area of networked health information. PMID- 8708261 TI - Inconsistencies in the literature on collegiate adult children of alcoholics: factors to consider for African Americans. AB - In a review of more than 50 literature entries on Adult Children of Alcoholics, the author found a number of inconsistencies. She emphasizes that clinicians and those who develop alcohol prevention programs should stop believing that certain characteristics are typical of all adult children of alcoholics. She makes specific recommendations for factors to be considered in programs for African American collegiate children of alcoholics. PMID- 8708262 TI - The Storey of college health hygiene. Thomas A. Storey, MD (1875-1943) and the promotion of hygiene. AB - The word hygiene originally defined a comprehensive plan for preserving individual and community health in all its dimensions. In the latter half of the 19th century, Dr Edward Hitchcock Jr established a system of hygiene at Amherst College that became the model for campus hygiene programs. Dr Thomas A. Storey of Stanford University, an advocate of teaching hygiene to college students, wrote articles and textbooks and was active on national committees that promoted college hygiene programs. Storey was one of the founders of the American Student Health Association, later renamed the American College Health Association, which in its early years chose promotion of hygiene as a major objective. As the 20th century progressed, the profession of health education emerged. With its emergence came the realization that health education, once viewed as primarily a matter of delivering health information to students, required an appreciation of human behavior as well. By 1950, the word hygiene had given way to health in most contexts in recognition of this new paradigm. PMID- 8708263 TI - Communication tools and college health. PMID- 8708264 TI - Pemoline therapy in college students with attention deficit hyperactivity disorder: a retrospective study. AB - Pemoline, a dopamine agonist, is effective in children with attention deficit hyperactivity disorder (ADHD), but its efficacy in adults is unknown. The authors studied the efficacy and safety of pemoline, using retrospective chart review of treated students with ADHD over a 2-year period. Forty students met diagnostic and treatment criteria; pemoline was associated with much improved or very much improved Clinical Global Impression symptoms scores in 70% of the students during a treatment period of 14 or more days. Severity of illness scores dropped from 4.11 to 3.01 between baseline and subsequent evaluation. Nine evaluable patients had adverse events, most commonly headaches, insomnia, and decreased appetite. Five additional students, who failed to meet the treatment-duration criterion, terminated because of severe initial insomnia. The authors concluded that pemoline is effective and safe in students with ADHD and has a lower abuse potential than methylphenidate and dextroamphetamine, the other two widely used, structurally dissimilar compounds, but controlled studies may be necessary before any final conclusions are reached. PMID- 8708265 TI - Medication for attention deficit hyperactivity disorder: commentary and response. PMID- 8708266 TI - Hazard surveillance and injury control: a university-based pedestrian-safety project. AB - Hazards to pedestrians at 15 campus crosswalk sites were identified, using a data collection protocol the authors developed so that two-person teams of trained observers could record the volume of vehicular and pedestrian traffic at the sites. Findings from the study revealed the nature of certain pedestrian hazards on campus, with specific intersections found to be more dangerous for pedestrians than others. The data collected were analyzed to create practical and effective hazard-control and safety programs for student groups. PMID- 8708267 TI - Consult-a-nurse: an innovative plan that did not succeed. PMID- 8708268 TI - The Internet: an essential tool for college health networking. AB - With the advent of the Internet, college health professionals have available to them a new setting in which to network. The Internet provides opportunities for immediate communication with a large and diverse community of colleagues. Through both the Student Health Service (SHS) Discussion Group and numerous health related databases, the Internet serves as a powerful research tool for program development and improvement. Answers to queries about practice options, solutions for local problems, and support of innovative policies can be received almost instantaneously from college health professionals from across the United States and Canada. In this article, the authors summarize the history and current use of the SHS Discussion Group, provide a brief overview of Internet basics, and identify more advanced uses of the Internet by college health professionals. PMID- 8708269 TI - Join the electronic revolution now! PMID- 8708270 TI - Facial pain. PMID- 8708271 TI - Dental licensure revisited. PMID- 8708272 TI - Dental licensure. PMID- 8708273 TI - Denture adhesives. PMID- 8708274 TI - Pediatric restoration. PMID- 8708275 TI - A conservative approach to placing posterior composite resin restorations. AB - Composite resins have changed the nature of dental restorations. However, many dentists have not altered their cavity preparation process, relying largely on the same procedure used for amalgam restorations. The author proposes a more conservative procedure, retaining more sound tooth structure, for placing posterior composite resin restorations. PMID- 8708276 TI - Screening for periodontal disease: radiographs vs. PSR. AB - This study compares the evaluation of periodontal health using bone levels measured on periapical and bitewing radiographs with clinical parameters, such as periodontal screening and recording, gingival indexes, probing depths and attachment levels. The authors found that bitewing radiographs taken within six months of the clinical examination correlated better with the clinical data than older radiographs. However, they concluded that, overall, radiographs are not highly reflective of periodontal status. PSR scores, by contrast, showed significant associations with probing depths and attachment levels. The results support use of PSR as a screening tool for periodontal diseases. PMID- 8708277 TI - Are head, neck and facial injuries markers of domestic violence? AB - Few data exist regarding the relationship between the location of injuries and the presence of domestic violence. This study of 127 people at an inner-city hospital emergency department found that most patients had head, neck and facial injuries. Although only 23 percent of patients with such injuries were victims of domestic violence, 94.4 percent of victims of domestic violence had head, neck and facial injuries. The study results indicate that head, neck and facial injuries could be markers of domestic violence. PMID- 8708278 TI - Assessing the cutting efficiency of dental diamond burs. AB - This study measured the cutting efficiencies of 15 types of roundend tapered conventional and disposable diamond burs. The results showed that disposable diamond burs had cutting efficiencies that were comparable to those of conventional (multiuse) diamond burs; their use should reduce the risk of clinical cross-infection. PMID- 8708279 TI - Identifying acute pulpalgia as a factor in TMD pain. AB - An acute pulpalgia and temporomandibular disorders can produce many of the same symptoms. To illustrate identification of an acute pulpalgia as a component in TMD, the authors review the evaluation of 11 patients at a clinic that specializes in TMD treatment. During the evaluation, thermal testing and periodontal ligament anesthesia were used to identify the offending tooth. After receiving endodontic treatment or having the tooth extracted, patients reported either complete or partial relief of TMD symptoms. The authors provide questions that may help practitioners identify a tooth with an acute pulpalgia as a contributing factor to TMD symptoms and suggest a technique to confirm this diagnosis. PMID- 8708280 TI - Enamel erosion resulting from hydrochloric acid tablets. AB - This article documents a case of severe enamel erosion caused by chewing hydrochloric acid tablets over a five-year period. Dentists must be aware that highly erosive acids are obtainable without a prescription, in this case from a health food store. They must also be alert for signs of dental erosion and identify the source of the acid. PMID- 8708282 TI - Dental radiographs and dental caries: a challenge. PMID- 8708281 TI - Infection control in the dental office: has anything changed? AB - Using data collected in two studies, the authors compare infection control practices used in dental offices in 1988 and 1992. During that time, dentists increased their use of barrier protection but still asked and learned very little about their patients. This lack of communication hinders dentists' ability to deliver the best care possible. The authors recommend that dentists improve communication with their patients to obtain better information about their health status. PMID- 8708283 TI - The legal implications of clinical practice guidelines. AB - In the past several years, considerable attention has been focused on clinical practice guidelines. They are developed to foster improved clinical care and to increase clinical efficiency. The legal implications of developing and following guidelines are not yet clear. The author argues that properly formulated guidelines could reduce the burdens of preventing and resolving malpractice claims. PMID- 8708284 TI - Notes on a "silent disease'. PMID- 8708285 TI - Hold-harmless agreements. What every dentist needs to know. PMID- 8708286 TI - Dentistry and sport. Meeting the needs of our patients. PMID- 8708287 TI - Delirium and dementia; are they distinct? PMID- 8708288 TI - Spouse caregivers' attitudes toward obtaining a diagnosis of a dementing illness. AB - OBJECTIVES: To examine attitudes of spouse caregivers about the process of obtaining a diagnosis of a dementing illness, including perceived benefits and obstacles to obtaining a diagnosis and suggestions for improving the process. DESIGN: A mail survey of spouse caregivers of dementia patients followed by the Michigan Alzheimer's Disease Research Center. PARTICIPANTS: The 233 spouse caregivers of dementia patients who returned a completed questionnaire. RESULTS: More than three-fourths of the sample rated the following benefits of obtaining a diagnosis as very or extremely important: ruling out other causes of memory or behavior problems, allowing family members to get information about dementia, may qualify my spouse for drug treatment, and will allow me to plan for the future. Primary obstacles included the time required to obtain a diagnosis, the cost of procedures, lack of access to physicians trained to diagnose dementing illnesses, and not receiving a referral from a primary care physician (41.6%, 41.0%, 40.8%, and 24.1% of the sample, respectively, agreed or strongly agreed that the factor posed a barrier to obtaining a diagnosis). Content analysis of responses to two open-ended questions identified problematic or stressful aspects of obtaining a diagnosis and suggestions for making the process easier for the patient and family. CONCLUSION: Results suggest the need to provide an orientation to the diagnosis and assessment process for patients and family members, develop a training session for physicians to better prepare them to address the emotional needs of patients and families and to disclose the diagnosis in an informative and compassionate manner, and provide outreach education to primary care physicians to increase knowledge and awareness of dementing illnesses and to increase referrals to community-based services and health professionals able to address the concerns of family members. PMID- 8708289 TI - Falls in community-dwelling older persons. PMID- 8708290 TI - Effects of age, Helicobacter pylori, and NSAID usage on the upper gastrointestinal tract. PMID- 8708291 TI - Elevated cerebrospinal fluid tau levels: implications for the early diagnosis of Alzheimer's disease. PMID- 8708292 TI - Will aging of the population make Japan less productive? PMID- 8708293 TI - Hospice services. PMID- 8708294 TI - The use of performance-based assessment instruments in cognitively impaired older patients. PMID- 8708295 TI - Urinary incontinence responsive to desmopressin: a case report. PMID- 8708296 TI - Prolonged colonic pseudo-obstruction (Ogilvie syndrome) in an older woman resolved with conservative treatment. PMID- 8708297 TI - Apolipoprotein-E genotyping of diabetic dementia patients: is diabetes rare in Alzheimer's disease? AB - OBJECTIVES: To determine whether diabetes is rare in Alzheimer disease (AD) relative to other types of dementia and whether diabetics with dementia have a low frequency of the Apolipoprotein-E E4 genotype. DESIGN: Observational survey study. SETTING: An Irvine, California, outpatient dementia assessment center. PARTICIPANTS: A total of 123 patients with AD, 51 with vascular dementia, 57 with "mixed" vascular dementia and AD, and 34 with "other" dementias (non-vascular non AD). MEASUREMENTS: Demographic data; histories and evidence of diabetes, hypertension, heart disease, stroke; and Apolipoprotein-E genotype for 95 cases distributed across the groups. RESULTS: There were 15 diabetics in the sample (5.7%), all of whom had extensive vascular disease. Diabetes was rare in AD patients (0.8%) relative to vascular dementia (11.8%), mixed vascular/AD dementia (8.8%), and "other" dementia patients (8.8%). In addition, the E4 allele of apolipoprotein-E, associated with high risk for AD, was frequent in the AD group (71.4%), but in the diabetic group it was only as frequent as in the general population (38.5%). In the diabetics with E4, 60% (3/5) had mixed dementia. CONCLUSIONS: Diabetics with dementia rarely have AD except as a component of mixed dementia. Apo-E genotyping showed only average E4 allele frequency in diabetics compared with the high E4 frequency found in AD patients. However, mixed dementia in diabetics may be associated with the E4 allele, suggesting that close control of diabetes may be particularly important for those with E4 since they may be more likely than others to develop both diseases. PMID- 8708298 TI - Smoking and cognitive impairment: a population-based study. AB - OBJECTIVE: To test the hypothesis that there is an inverse relationship between smoking and cognitive impairment by examining the prevalence and incidence of such impairment in a representative cohort of older urban residents. DESIGN: Survey follow-up with three interviews over a 4-year period. SETTING: City of Cleveland, Ohio. PARTICIPANTS: Six hundred forty-seven survivors of a population based, multi-stage probability sample of urban residents, aged 74 years and older in 1984. MEASUREMENTS: Past and present cigarette smoking was measured in 1984 by direct questioning. Cognitive impairment was measured in 1984, 1987, and 1988 by the 10-item Short Portable Mental Status Questionnaire. Mortality was determined at each interview date, and death certificates were obtained. Data were analyzed by univariate analysis, stratified analysis, and multivariate logistic regression. RESULTS: There were 99 cases of cognitive impairment at baseline among 628 individuals for whom complete data were available. Crude data suggest that the prevalence of cognitive impairment was lower among smokers than among non-smokers, but logistic regression, adjusted for age, income, and gender, showed this difference to be non-significant, OR 0.73; 95% CI (0.42, 1.29). Fifty one new cases of cognitive impairment occurred during the 4-year observation period. Neither the crude data nor logistic regression showed any significant relationship between smoking and the incidence of cognitive impairment, OR 1.03; 95% CI (0.54, 1.99). Four-year mortality did not differ significantly between these older smokers and non-smokers, but smokers died at a relatively younger age and were more likely to die of diseases associated with smoking. PMID- 8708300 TI - Geriatric targeting criteria as predictors of survival and health care utilization. AB - OBJECTIVE: To assess the utility of geriatric targeting criteria in predicting survival and health care utilization in a cohort of hospitalized older veterans. DESIGN: A prospective cohort study assessing geriatric targeting criteria, e.g., polypharmacy, falls, or confusion, with respect to adverse outcomes at 12 months. SETTING: A Tertiary Care VA Medical Center. PATIENTS: 507 acutely hospitalized male veterans aged 65 years or more. MAIN OUTCOME MEASURES: Survival status, nursing home placement, and total hospital days during 12 months following hospital admission. RESULTS: Patients who had a higher number of targeting criteria at admission showed a significantly increasing trend toward death (P < or = .001), nursing home placement (P < or = .01), and longer hospital stays (P < or = .01) at 12 months. In univariate analyses, weight loss (relative hazard 3.8, 95% CI 2.4, 5.9), appetite loss (relative hazard 3.3, 95% CI 1.9, 5.8), depression (relative hazard 2.5, 95% CI 1.4, 4.5), falls (relative hazard 2.2, 95% CI 1.2, 4.1), confusion (relative hazard 2.2, 95% CI 1.2, 4.0), and socioeconomic problems (relative hazard 1.6, 95% CI 1.0, 2.5) predicted death. Polypharmacy (OR 3.4, 95% CI 1.3, 8.8), confusion (OR 4.4, 95% CI 1.5, 13.0), and prolonged bedrest (OR 7.6, 95% CI 1.5, 39.3) predicted nursing home placement. Confusion (Beta 12.0, 95% CI 2.9, 21.3), falls (Beta 14.2, 95% CI 4.2, 24.3), and prolonged bedrest (Beta 22.4, 95% CI 3.9, 41.0) predicted total hospital days. In multivariate analyses, weight loss, depression, and socioeconomic problems predicted death; confusion and polypharmacy predicted nursing home placements; and falls predicted total hospital days. CONCLUSION: This prospective cohort study of hospitalized older veterans demonstrated geriatric targeting criteria as predictors of adverse hospital outcomes. Our findings suggest screening acutely hospitalized patients using chart abstracted geriatric targeting criteria is useful in identifying patients at risk for adverse outcomes of hospitalization. PMID- 8708299 TI - Report of an outbreak: nursing home architecture and influenza-A attack rates. AB - OBJECTIVE: To determine factors that might account for a significantly lower attack rate in a newly constructed nursing building during an epidemic of type A influenza. SETTING: A four-building, long-term care facility for veterans and their spouses, with an average daily census of 690. DESIGN: Prospective surveillance with retrospective analysis. PARTICIPANTS: Symptomatic residents submitting to viral culture. MEASUREMENTS: Number of respiratory illnesses and influenza cultures in consenting symptomatic residents. Building characteristics. RESULTS: An influenza A (H3N2) outbreak was culture-confirmed in 68 nursing home residents. Influenza A was isolated in 3/184 (2%) residents in Building A, 31/196 (16%) in Building B, 18/194 (9%) in Building C, and 16/116 (14%) in Building D. Denominators are average daily census during the outbreak. Building A had significantly fewer culture-confirmed cases than the other buildings (P < .001). Fewer residents in Building A, 47% compared with 61% in Buildings B, C, and D, were participants in a formal study of influenza. Eight of 15 respiratory illnesses identified during the outbreak that were not cultured occurred in Building A. These factors could not account for the difference in attack rates. Building A has a unique ventilation system, more square feet of public space per resident, and does not contain office space that serves the entire four-building facility. CONCLUSION: Our retrospective observation suggests that architectural design may influence the attack rate of influenza A in nursing homes. PMID- 8708301 TI - Variability in mammography use among older women. AB - OBJECTIVE: To determine rates of and explore factors associated with mammography use among older women. DESIGN: Retrospective review of part B (physician) bills submitted to Medicare during 1990. SETTING: Health Care Financing Administration (HCFA) data, including sociodemographic information and part B physician bills for all services delivered to Medicare-eligible women in 1990. PATIENTS/PARTICIPANTS: Women age 65 or older as of January 1, 1990, residing in one of 10 states with part B coverage through December 31, 1990. MEASUREMENTS AND MAIN RESULTS: The outcome was receipt of a mammogram (yes/no). We explored factors associated with mammography use within three age groups: 65 to 74, 75 to 84, and 85+. The factors considered were race, state, median income of ZIP Code of residence (from the 1990 US Census, and used to divide the population into quintiles within each state), and number of primary care visits (0, 1, 2, and 3+). Overall, 15% of women had a mammogram: 20% of women age 65 to 74, 12% of women age 75 to 84, and 4% of women age 85 and older. Mammography use was lowest in Oklahoma and highest in Washington. However, in each state the older the age category, the less the mammography use (e.g., 9% vs 5% vs 2% in Oklahoma and 25% vs 16% vs 5% in Washington for women 65-74, 75-84, and 85+, respectively). Mammography use was lower for black than for white women age 65 to 74 (14% vs 21%, P < .001) and 75 to 84 (9% vs 12%, P < .001). Women in each of these two age groups had lower mammography use if they resided in the lowest income quintile and highest if they resided in the highest income quintile (17% vs 23% 65-74, and 10% vs 13% 75-84, P values < .001). Among the oldest women (those 85+), mammography use was low (4%) and varied minimally by race and income (P = .907 and .003, respectively). In all age groups, mammography use was lowest among women who did not have a primary care visit, was greater among women who had at least one visit, and continued to rise with increasing numbers of visits (all P values < .001). For example, among women age 75 to 84, mammography use increased from 5% to 10%, 14%, and 17% for those with 0, 1, 2, and 3+ visits. CONCLUSIONS: We found that mammography use was less for women who were older, of black race, who did not visit a primary care provider, and who lived in areas with lower median income and certain geographic locations (states). Similar factors influenced mammography use in women age 65 to 74, where there is greater consensus as to who should receive a mammogram, and women age 75 to 84, where there is neither consensus nor data. Surprisingly, neither race nor income had much influence on mammography use among women age 85 or older. PMID- 8708302 TI - Bacteremic urinary tract infection in older people. AB - OBJECTIVES: To describe patients with bacteremic urinary tract infections, compare characteristics of young and older patients, and suggest appropriate empiric antibiotic therapy for this clinical condition. DESIGN: Case series, with an observation period of 3 years. SETTING: A large southeastern community hospital. PARTICIPANTS: One hundred eighty patients more than 18 years old (101 older than age 65) with urine and blood cultures simultaneously positive for bacterial organisms. MEASUREMENTS: Chart review-determined demographic characteristics, clinical data, bacterial organism and antibiotic susceptibility, number of medical diagnoses, use of a urinary catheter, admission source, and mortality. RESULTS: The 180 patients experienced bacteremias caused by 183 bacterial organisms. Sixty-one patients were aged 65 to 79 years, and 40 were 80 years of age or older; 63.9% of patients were female; 62.2% were black. Gram negative organisms accounted for 80.3% of bacterial isolates, with Escherichia coli accounting for 54.1% of cases. Gram-positive organisms accounted for 19.7% of isolates, including Staphylococcus aureus (13.1%) and Enterococcus (5.5%). Older patients had a distribution of Gram-positive and Gram-negative organisms similar to that of younger patients. Men and patients with chronic urinary catheters had higher proportions of Gram-positive organisms and non-E. coli Gram negative rods. Organisms showed excellent susceptibilities to commonly used antibiotics. In-hospital mortality was 16.1%, with higher mortality in patients with chronic urinary catheters, in patients who were admitted from nursing homes, and when a Gram-positive organism was identified. Advanced age was not associated with higher mortality. CONCLUSIONS: Concomitant illness and especially the use of urethral catheters, but not advanced age itself, are associated with a higher mortality from bacteremic urinary tract infection. Single agent empiric antimicrobial therapy such as ceftriaxone may be appropriate in older patients with presumed urosepsis, except in catheterized patients or those with other risk factors for Gram-positive or resistant Gram-negative infections. PMID- 8708303 TI - How older patients' treatment preferences are influenced by disclosures about therapeutic uncertainty: surgery versus expectant management for localized prostate cancer. AB - OBJECTIVES: There is an ongoing debate about the proper management of localized prostate cancer in older men. We assessed whether older patients differ in their willingness to accept surgery versus expectant management for a hypothetical early stage, low grade, clinically localized prostate carcinoma, and whether patients' current levels or past history of urologic dysfunction (difficulties initiating urination, failing to empty the bladder, urinary dribbling, getting up at night to urinate, and frequency of sexual activity) influence their decisions. We assessed patients' willingness to choose surgery over expectant management by varying the expected survival benefit in years (ESBs-em) of surgery over expectant management. DESIGN: Structured interviews with a consecutive series of male patients. SETTING: A university-based Department of Veterans Affairs Medical Center. PATIENTS: One hundred forty-eight patients seen consecutively in General Medicine Clinic at the Department of Veterans Affairs Medical Center in Portland, Oregon, were enrolled in the study. Mean age of the patients was 66.3 years (SD = 10.3, range = 30-85); mean level of formal education was 12.6 years (SD = 2.7, range = 6-22). MEASUREMENTS: Patients were asked whether they would accept surgery or expectant management in one of 11 treatment comparisons. We varied the ESBs-em in 1-year increments from 0 years to 10 years. As described to patients, surgery carried a mortality risk at the time of treatment of 1 to 2%. Once any patient indicated a willingness to accept surgery at any of the treatment comparisons or if any patient reported preferring expectant management across all treatment comparisons, the elicitation procedure was stopped. All patients were asked to complete a urological and sexual functioning questionnaire to determine the presence of coexisting urological dysfunction and level of sexual activity both at present and in the past. RESULTS: Of the 148 patients enrolled in the study, 43.2% (64/148) preferred surgery with a zero expected life benefit over expectant management (ESBs-em = 0) and a 1 to 2% chance of dying within 1 month of surgery; 24.3% (36/148) rejected surgery as the expected life benefit of surgery was increased (0 < ESBs-em < or = 10 years); 26.4% (39/148) preferred expectant management even when there was a 10-year expected life benefit of surgery; 4.7% (7/148) preferred that their physician make the decision for them; and 1.4% (2/148) of patients reported that they preferred radiation therapy, an option that was not offered to them explicitly. Our results suggest that older patients are more likely to report a preference for expectant management (OR = 1.07). Further, our results suggest that patients who report current urinary dribbling (OR = 9.03) are much more likely to prefer expectant management but that this preference decreases with the amount of time they have had this problem. Similarly, we find that patients who have difficulty with starting urination are much more likely to prefer surgery (OR = 0.13), and this preference is also mediated by the number of years they have experienced this problem. Treatment choice was not associated with formal education, present health status, or the other urological symptoms we assessed. CONCLUSION: Our study in an older male veteran population showed preferences for a variety of options in prostate cancer. Although the majority of men preferred surgery, a significant number preferred expectant management. Our results show that preferences reflect patients' experiences with physical problems associated with disease and that these experiences need to be explored and considered by patients and their providers when making treatment decisions. PMID- 8708304 TI - The relationship between ethnicity and advance directives in a frail older population. AB - OBJECTIVE: To assess the relationship between ethnicity and Health Care wishes, including Advance Directives, in a group of frail older persons in PACE (Program For All Inclusive Care Of The Elderly). DESIGN: Retrospective chart review of 1193 participants in the PACE program. SETTING: Program of All Inclusive Care Of The Elderly (PACE), a comprehensive managed care demonstration program serving frail older participants at 10 sites across the nation. PARTICIPANTS: A total of 1193 older adults, all of whom met state criteria for nursing home level of care. There were 385 non-Hispanic whites, 364 blacks, 156 Hispanics, and 288 Asians. MEASUREMENTS: Presence or absence of advance directives, type of health care wishes selected including living will, durable power of attorney, and health care proxy. RESULTS: Frail older white, black, Hispanic and Asian Americans differ significantly in their health care wishes and how they choose to express them. Blacks were significantly more likely to select aggressive interventions and less likely than non-Hispanic whites and Hispanics to utilize a written instrument for expressing health care wishes. Whites were significantly more likely to utilize written documents for advance directives, whereas Asians were more likely to select less aggressive interventions but were unlikely to use written advance directives. CONCLUSIONS: In this population, we found significant ethnic variations in choice of health care wishes. Although health care wishes are an individual decision, an awareness of cross cultural patterns can assist practitioners in addressing the concerns of their patients, as well as assisting Health Care Policy Development. PMID- 8708305 TI - Drug-drug interactions related to hospital admissions in older adults: a prospective study of 1000 patients. AB - OBJECTIVE: To investigate the frequency, nature, and side effects of drug-drug interactions (DDI) in a group of geriatric inpatients. DESIGN: Study of drugs administered at home in the 2 weeks preceding hospitalization. SETTING: Evaluation of patients admitted to a geriatric unit from the emergency unit. PATIENTS: One thousand inpatients more than 70 years of age (83.0 +/- 7.1 years), admitted consecutively to a hospital for acute illness. MEASUREMENTS: All possible two by two combinations of drugs administered at home were considered to determine whether these associations could lead to a DDI and whether side effects resulted from these interactions. RESULTS: Five hundred thirty-eight patients were exposed to 1087 DDI. The most frequently involved drugs were cardiovascular and psychotropic medications. There were 189 side effects observed in 130 patients. The most frequent side effects were neuropsychological impairment, arterial hypotension, and acute renal failure. The number of side effects did not differ between the 66 contraindicated drug associations and the 1021 associations that only required precautionary use. CONCLUSIONS: DDI frequently lead to side effects in older adults. Classifications of DDI must be adapted to increased DDI sensitivity in this population. Nevertheless the frequency of side effects might be reduced by limiting the prescription of the most frequent and dangerous DDI. PMID- 8708306 TI - Free and protein-bound cobalamin absorption in healthy middle-aged and older subjects. AB - OBJECTIVE: To study free- and protein-bound cobalamin absorption and the correlation with atrophic gastritis in healthy middle-aged and older subjects. DESIGN: A cross-sectional study. PARTICIPANTS: Fifty-two healthy subjects, aged 26 to 87 years, apparently free from conditions known to influence the cobalamin status. Middle-aged subjects were defined as those younger than 65 years of age (median age 57 years) and older subjects as those 65 years and older (median age 75 years). MEASUREMENTS: Protein-bound cobalamin absorption was assessed by 48 hour urinary excretion method following oral administration of scrambled egg yolk, labeled in vivo with 57 Co-cobalamin by injecting a hen with 57 Co cyanocobalamin. The percentage of 57 Co-cobalamin bound to protein was 65%. Free cobalamin absorption was assessed by 48-hour urinary excretion method following oral administration of crystalline 57 Co-cyanocobalamin. Plasma cobalamin, folate and fasting plasma gastrin, and pepsinogen A and C concentrations were determined. RESULTS: The median urinary excretion of egg yolk 57 Co-cobalamin in middle-aged subjects was 12.3% (25th and 75th percentiles 10.5%-14.5%) compared with 11.7% (25th and 75th percentiles 9.8%-13.6%) in older subjects (P = .283). The median urinary excretion after administration of free 57 Co-cobalamin in middle-aged subjects was 25.7% (25th and 75th percentiles 20.6%-30.7%) compared with 27.9% (25th and 75th percentiles 21.4%-34.5%) in older subjects (P = .694). Neither egg yolk nor free 57 Co-cobalamin excretion correlated with age. A ratio of pepsinogen A to pepsinogen C less than 1.6, indicating atrophic gastritis, was found in 13 subjects. Within the atrophic gastritis group, 11 subjects had a pepsinogen A concentration greater than or equal to 17 micrograms/L, indicating mild to moderate atrophic gastritis, and two subjects had a pepsinogen A concentration less than 17 micrograms/L, indicating severe atrophic gastritis or gastric atrophy. All subjects had normal fasting plasma gastrin concentrations. Free and egg yolk 57 Co-cobalamin excretions were not reduced in the atrophic gastritis group when compared with the non-atrophic gastritis group. Median plasma cobalamin concentration was not significantly lower in older subjects (P = .205). Nonetheless, plasma cobalamin concentration correlated negatively with age (r = -.36; P = .008). CONCLUSIONS: We demonstrated no significant difference in either free or protein-bound cobalamin absorption between healthy middle-aged and older adults. In addition, no alteration in cobalamin absorption was found in subjects identified as having mild to moderate atrophic gastritis. Therefore, based on our results, the high prevalence of low cobalamin levels in older people cannot be explained by either the aging process or mild to moderate atrophic gastritis. PMID- 8708307 TI - Life values, resuscitation preferences, and the applicability of living wills in an older population. AB - OBJECTIVES: To determine whether life values are related to resuscitation preferences and living will completion in an older population and to assess beliefs about the applicability of living wills. DESIGN: Individual structured interviews. SETTING: An independent retirement community. PARTICIPANTS: One hundred thirty-two subjects older than 63 years of age. MEASUREMENTS: Resuscitation preferences were elicited in five hypothetical scenarios. Subjects with living wills were asked whether their living will would play a role in the scenarios. Subjects rated the importance of 13 life value statements. RESULTS: The percentage of subjects desiring CPR in each scenario was as follows: current condition (66%); acute illness (33%); terminal disease (8%); functional impairment (8%); and dementia (7%). The percentage of those with a living will who thought their living wills would play a role in the scenarios was as follows: acute illness (84%); terminal disease (93%); functional impairment with intact cognition (66%); and dementia (91%). Factor analysis of the life value statements revealed five meaningful factors: quality of life; capacity/autonomy; family relations; physical comfort; and treatment philosophy. Multiple correlations were found between four of five life value factors and hypothetical resuscitation preferences or the presence of a living will. CONCLUSION: Subjects misinterpreted the applicability of living wills in nonterminal illness scenarios. A relationship between life values and resuscitation preferences was noted, which emphasizes the importance of eliciting and including life values when discussing advance directives. PMID- 8708309 TI - Characteristics of urinary incontinence in homebound older adults. AB - OBJECTIVE: To describe the characteristics of urinary incontinence and related factors in incontinent homebound older adults. DESIGN: A descriptive study of 90 cognitively intact incontinent homebound older persons referred to a clinical trial to examine the effectiveness of behavioral therapies in the treatment of urinary incontinence (UI) in homebound patients. SETTING AND PARTICIPANTS: Incontinent patients more than 60 years of age who met the Health Care Financing Administration's definition of homebound were referred to a clinical trial by home care nurses from a Medicare-approved home care agency in a large metropolitan country in Pennsylvania. MEASURES: Structured continence and medical history, basic and instrumental activities of daily living, Folstein MMSE, Geriatric Depression Scale, mobility-toileting skills, bladder diaries, and physical examination. RESULTS: Four hundred eighty-four persons were referred to the clinical trial, and 90 cognitively intact persons were found eligible to participate in this study (80 women and 10 men). Subjects had a mean age of 75.8 years, reported a mean of 8.4 medical problems, and most, 80%, had functional limitations in ambulation. Subjects recorded a mean of 3.8 urinary accidents/day 1.4 large and 2.4 small accidents/ day-in baseline bladder diaries. The majority, 73.3%, had more than 10 accidents per week, and most patients reported mixed urge, stress (57.1%), or pure urge (37.7%) UI. Half (54.4%) reported that UI further restricted their activities, and 52.2% reported that this problem was extremely disturbing. However, 90.5% believed that UI could be treated. CONCLUSIONS: Urinary incontinence tends to be severe among cognitively intact homebound older adults in both frequency and volume of accidents. Although subjects were homebound with many health and functional disabilities, they perceived UI as a very disturbing problem that further restricted their activities. Participants in this study were optimistic about the potential benefits of treatment. PMID- 8708308 TI - Nutritional risk in inner-city-dwelling older black Americans. AB - OBJECTIVE: To define the degree of nutritional risk in older inner-city black Americans and to identify important underlying factors associated with high nutritional risk. DESIGN: Cross-sectional descriptive study. SETTING AND PARTICIPANTS: A population-based sample of 400 noninstitutionalized persons older than 69 years of age in north St. Louis (NSL), Missouri, and a community-based sample of 115 residents aged 50 years and older living in public housing in East St. Louis (ESL), Illinois. Both study areas have high levels of poverty. MEASUREMENTS: Nutritional risk was measured using the Nutrition Screening Initiative Checklist. Demographic information, economic status, self-rated health, Geriatric Depression Scale score, and body mass index were assessed using established standardized instruments. MAIN RESULTS: Forty-eight percent of NSL and 66% of ESL subjects scored high on the Checklist. Compared with a mostly white (96%) comparison group from New England, both samples demonstrated particularly high prevalence for limited intake of fruits, vegetables, and milk; tooth and mouth problems; lack of money for food; eating alone; polypharmacy; and inability to shop, cook or feed on their own. High levels of depressive symptoms, fair or poor self-rated health, perceived inadequacy of income, and low income levels were associated with high risk, but even those subjects with no or few such predisposing factors were still high on the Checklist score compared with the New England sample. CONCLUSIONS: If confirmed, these results indicate that inner-city-dwelling older black Americans are at high nutritional risk. Attempts to reduce their nutritional risk should focus on improving nutritional content of their diet, oral health, polypharmacy, depressive symptoms, and poor general health; offering group meals; and providing assistance with shopping and cooking. PMID- 8708310 TI - Utility of clysis for hydration in nursing home residents. AB - OBJECTIVES: To determine the utility of hypodermoclysis in the management of dehydration in nursing home residents and to review the recent literature on the use of clysis in older adults. DESIGN: A retrospective chart review. SETTING: Two continuing care retirement community nursing home units in Cincinnati. PARTICIPANTS: Thirty-six nursing home residents with a mean age of 85 years. MEASUREMENTS: A standard chart auditing form was used to collect demographic and clinical data from the patients' nursing home charts. Clinical outcomes were assessed at 1 week following the cessation of clysis. RESULTS: The study subjects were functionally impaired, with 86% having significant cognitive impairment and 81% totally dependent with transfer. The most common indication for clysis was in association with an infection. Clysis was frequently initiated upon nursing observation of inadequate oral intake before obtaining laboratory results and discontinued upon resumption of adequate oral intake. The use of clysis was associated with the return to clinical/functional baseline in 71% of the cases. In the subset of subjects with pre- and post-clysis measurements of serum sodium and BUN/creatinine ratios, improvement in laboratory values was not significant. The use of clysis was not associated with significant complications, but in nine cases minor local skin reactions were documented. CONCLUSIONS: Hypodermoclysis is a relatively safe and effective procedure in a nursing home. The use of clysis in the nursing home is an alternative to intravenous hydration. The use of clysis for short-term hydration has the potential to reduce cost and transfers to the hospital. PMID- 8708311 TI - Hand sensibility of healthy older people. AB - OBJECTIVE: To develop normative data for four hand sensibility modalities in older subjects. DESIGN: Cross-sectional. SETTING AND PARTICIPANTS: Three hundred and sixty community-dwelling subjects of both sexes, aged 60 to 94, randomly selected from the electoral list of the city of Sherbrooke, Quebec, Canada. MEASUREMENTS: Touch/pressure threshold (Semmes-Weinstein monofilaments), static and moving two-point discrimination (Mackinnon-Dellon Disk-Criminator), tactile recognition (Modified Pick-up test), and thumb kinesthesia. RESULTS: A reduction with age was found in the performance of the study subjects, with the exception of the kinesthesia test. The values obtained in this study are clearly lower than the norms proposed for adults, underlining the importance of using reference values developed for the target clientele. CONCLUSION: The norms will help clinicians to differentiate better between normal and pathological changes in sensibility with age. PMID- 8708312 TI - D-E-N-T-A-L: a rapid self-administered screening instrument to promote referrals for further evaluation in older adults. PMID- 8708313 TI - Multiple surgery indications in an older woman undergoing stroke rehabilitation: the rationale of complex decision-making in geriatric medicine. PMID- 8708314 TI - Making treatment decisions for incapacitated older adults without advance directives. AGS Ethics Committee. American Geriatrics Society. PMID- 8708315 TI - Longevity assurance genes: how do they influence aging and life span? PMID- 8708316 TI - Alzheimer's disease and non-insulin-dependent diabetes mellitus: common features do not make common bedfellows. PMID- 8708318 TI - Oral health for the older patient. PMID- 8708317 TI - Challenges facing physicians and healthcare institutions caring for patients with mental incapacity. PMID- 8708319 TI - Mental health services research on the aging: new opportunities and challenges for clinical researchers. PMID- 8708320 TI - What can you do about your patient's dry skin? AB - Water, absorbed through bathing, will hydrate skin but will rapidly be lost to evaporation unless a topical occlusive agent is applied to prevent moisture loss through the skin. The frequency of bathing (at least 5 times or more weekly) is more important than the mode of bathing (tub vs. shower) in effectively treating dry skin. In addition to the elements critical to prevention of dry skin through bathing practices, environmental humidity and fluid intake must be therapeutic. PMID- 8708321 TI - Hope, chronic illness and the elderly. PMID- 8708322 TI - Communication difficulties of nursing home residents. How can staff help? AB - Recent studies have found a high prevalence of communication disorders in elderly residents of nursing homes. This study was undertaken to investigate staff attitudes to the communication problems and communicative needs of residents in their care. A questionnaire was administered to 32 staff members from one nursing home. Responses indicate little evidence of the negative staff attitudes previously reported in the literature. Staff wanted more time to spend communicating with residents and wanted to learn more about the residents' communication problems. Implications of the questionnaire responses for staff training are presented. PMID- 8708323 TI - Managing ambiguous loss in dementia and terminal illness. AB - Grieving ambiguous loss is difficult because individuals must let go while finding ways to remain connected. In cases of ambiguous loss, nurses can play an important role in helping bereaved families through empathetic "presencing" and objective coaching. Coaching roles include helping family members grieve openly by sharing perceptions, and problem solve together to manage individual, caregiving and broader family issues. Nurses provide hope that a disease can be managed by making the unfamiliar familiar. PMID- 8708324 TI - Guideline for prevention and control of antibiotic-resistant microorganisms in california long-term care facilities. Centers for Disease Control. PMID- 8708325 TI - "Isolation" in the nursing home setting. PMID- 8708326 TI - Rotavirus-associated diarrhoea in children in a refugee camp in Jordan. AB - Studies on the rotavirus-associated acute diarrhoeal illness in Jordanian children are non-existent. The present case-control study was conducted to investigate the prevalence of rotavirus diarrhoea among children aged less than 5 years, attending the United Nations Refugee World Aid Clinic in northern Jordan. The potential environmental and behavioural risk factors contributing to the infection were also studied. Using the ELISA technique rotavirus antigens were detected in the stool samples of 35% of the 220 cases of gastroenteritis and in 3% of the control group. The control subjects were matched for age and sex with the cases. The overall prevalence was significantly higher (62%) in children aged less than 24 months [OR = 2.4, 95% CI (1.1-5.1)] than those in the older age groups. Severe cases of diarrhoea were rare. Diarrhoea due to rotavirus was more prevalent during the summer months (June-August). Risk factors for acute diarrhoea in these children are related to the infant feeding practices of using unboiled tap water to prepare the formula milk, and the low educational level of the mothers. PMID- 8708327 TI - Molecular epidemiology of nosocomial rotavirus infection. AB - The molecular epidemiology of nosocomial rotavirus infection in children admitted to the paediatric ward of a large hospital in southern India was studied by transmission electron microscopy and RNA electrophoresis on stool samples. Six hundred and twenty-six samples were collected and rotaviruses were detected in 35 of them. A predominant electropherotype was found in all the samples, and there was no seasonal variation in the incidence of infection. The predominant electropherotype pattern in this study was different from the electropherotype patterns of strains isolated elsewhere in the country and also from the pattern of the strains isolated earlier from children with acute diarrhoea at the same hospital. PMID- 8708328 TI - Protection against mortality due to Vibrio cholerae infection in infant rabbits caused by immunization of mothers with cholera protective antigen. AB - Vaccination of female rabbits with cholera protective antigen (CPA) protected their F1 progeny from lethal challenge with Vibrio cholerae. Protection was determined by the choleragenic score and survival rates. Serum and milk IgG, IgM, IgA titres to CPA, cholera toxin, and LPS were determined. At 8 and 20 weeks post immunization, mothers' milk, sera, and infants' sera showed elevated CPA-specific IgG and IgA, and infants were protected. Mothers' serum and milk antibody remained elevated for 36 weeks. At 26 weeks, mothers were re-bred, but their progeny were swapped and cross-fed. Infants born to the placebo-vaccinated mothers and nursed by CPA-immune nannies were partially protected from challenge. Infants born to CPA-immune mothers and cross-fed by the placebo-vaccinated nannies were less protected. CPA stimulated both transplacental and milk antibody, but passive immunity was primarily milk-derived. A 36-week booster vaccine stimulated an anamnestic serological response that did not provide protection equivalent to the original vaccine. CPA provided partial protective immunity to the milk-fed infant rabbits that suggests that CPA may be important in the development of a cholera vaccine. PMID- 8708329 TI - Colonization of streptomycin-treated mice by Aeromonas species. AB - Streptomycin-treated adult mice were investigated as a possible model for studying the enteropathogenicity of Aeromonas species. C57BL mice pre-treated with streptomycin (5.0 g/L drinking water, 48 hours) received a single intragastric dose (10(10) bacteria /10.5 mL) of one of six well-characterized, toxin-producing, human diarrhoeal isolates of A. veronii biovar sobria (n = 3) or A. hydrophila (n = 3). Their faeces were examined for Aeromonas for 10 days post challenge. All strains colonized the antibiotic-treated mice. Colonization did not occur in mice which did not receive streptomycin. Strains of A. hydrophila were recovered in greater numbers than strains of A. veronii biovar sobria, and colonized ( > or = 10(3) cfu/g of faeces) a greater proportion of mice at day 10. Strains of the latter species, however, were more adherent in cell line assays used as models of intestinal adhesion. A. hydrophila strains localized in the large intestine and appeared not to be cell associated. This study, therefore, points to species-related differences in intestinal colonization mechanisms. The streptomycin-treated adult mouse model may prove useful for further investigation of some of these mechanisms. Diarrhoeal symptoms were, however, not produced in this model. PMID- 8708331 TI - Survival of Shigella flexneri in artificial aquatic environment: effects of different physicochemical stress factors. AB - Survival of bacteria in aquatic environment is influenced by the prevailing physicochemical characteristics of water. A study was carried out to assess the effects of temperature, pH, and salinity of water on the survival of Shigella flexneri in laboratory microcosms. Survival of S. flexneri was assessed by viable counts of Shigella on MacConkey agar plates. At pH 7.0 and temperature of 4 degrees C, S. flexneri survived longer in 0.5% salinity than in salt-free distilled water, while at pH 7.0 and at higher temperatures (25 degrees C and 37 degrees C), the bacteria survived longer in distilled water than in 0.5% salinity. It was also observed that 2.0% NaCl solution was detrimental to bacterial survival at pH 8.0 and temperature of 37 degrees C. Survival times of the viable cells increased with reduction of temperatures and salinity. These findings demonstrated that survival of S. flexneri in the aquatic environment is greatly influenced by physicochemical factors. PMID- 8708330 TI - Knowledge, attitude, practice, and prescribing pattern of oral rehydration therapy among private practitioners in Nigeria. AB - To determine the knowledge, attitude, and practice of oral rehydration therapy (ORT) among private medical practitioners in Enugu, Nigeria, 91 doctors were interviewed using a structured questionnaire. All the doctors had heard of ORT and believed in its efficacy. The commonest source of information on ORT was the medical school (44%). Fifty percent would recommend salt-sugar solution (SSS) rather than oral rehydration solution (ORS). The main reason is its cost effectiveness and easy availability. Only 55% of the respondents knew how to prepare SSS correctly. The percentage of doctors who prescribe smooth muscle relaxant (spasmolytic use rate) was 41%, and the commonest reason for its use was to reduce bowel movement. The influence of year of medical graduation on spasmolytic use was found to be statistically significant (p < 0.05). Antibiotics were commonly used, although most (76%) doctors believed that viral infections were a common cause of childhood diarrhoea. All the respondents would recommend continued breastfeeding during diarrhoeal episodes. The study revealed a high rate of inappropriate drug use and a deficiency in the knowledge and practice of ORT. PMID- 8708332 TI - Anti-Plesiomonas shigelloides agglutinating and complement-fixing antibody titres in normal individuals and diarrhoeal patients in Edo State, Nigeria. AB - One hundred patients with diarrhoea and 50 asymptomatic individuals attending various hospitals in Edo State, Nigeria, were screened for serum complement fixing and agglutinating antibodies to Plesiomonas shigelloides using the complement-fixation and agglutination tests. Seventy (70%) of the 100 patients and 20 (40%) of the 50 asymptomatic individuals had detectable complement-fixing antibodies at titres ranging from 1:32 to 1:128 and 1:8 to 1:32 respectively. Results suggest that cases of diarrhoea in this environment may be due to P. shigelloides, but the demonstration of antibodies in asymptomatic individuals show that they also have serum antibodies against P. shigelloides. The exclusive use of antibody responses in the diagnosis of P. shigelloides infections should, therefore, be interpreted with caution. PMID- 8708333 TI - Isospora belli infection in a patient with acute lymphoblastic leukaemia in India. PMID- 8708334 TI - Faecal leucocytes in cholera due to Vibrio cholerae O139 Bengal. PMID- 8708335 TI - Bibliography on diarrhoeal diseases. PMID- 8708336 TI - Promotion of breast feeding in the community: impact of health education programme in rural communities in Nigeria. AB - Breast feeding has been recognized as a child survival strategy, while breast feeding programmes have been increasingly implemented in many communities. This study assesses the effectiveness of a breast feeding education programme launched through the primary health care programme in the rural communities of Nigeria. Late trimester pregnant women were enrolled into the study and given a questionnaire on knowledge, attitudes, and practices (KAP) about breast feeding. Women in the study group (n = 126) received breast feeding counselling before and after delivery, while those in control group (n = 130) did not receive any counselling. Both groups were monitored after delivery and followed with the KAP questionnaire. The results of the study showed marked improvements in the intervention group for colostrum feeding (p = 0.0000). Moreover, 31.6% of the mothers in the intervention group practised timely initiation of breast feeding compared to 5.6% of the controls, and the prevalence of exclusive breast feeding at 4 months was 39.8% in the intervention group compared to 13.9% for the controls. Multivariate analysis showed that the intervention was a powerful and the only significant predictor of the increase in breast feeding behaviours (p = 0.0000), and that an early initiation of breast feeding is a strong predictor of exclusive breast feeding at 4 months of age. It is concluded that breast feeding promotion in rural communities is feasible and can lead to behavioural changes. PMID- 8708337 TI - Thinking, talking, acting. PMID- 8708338 TI - Talking heads and sleeping citizens: health policy making in a democracy. PMID- 8708339 TI - Lawmakers' views on the failure of health reform: a survey of members of Congress and staff. PMID- 8708340 TI - Medical malpractice and no-fault systems. PMID- 8708341 TI - Publicity-stunt participation and sound bites polemics: the health care debate 1993-94. AB - The outcome of Bill Clinton's health care initiative presents policy analysts and democratic theorists with a puzzle: How could vigorous discussion of health reform, introduced into the context of an emerging public consensus about its urgency, produce widespread incomprehension of the issue and foster public reluctance to embrace any specific proposal for change? Starting from the perspective of political theories of deliberative democracy, I approach the puzzle of failed health reform by pointing to the paradox of decision making in an adversary democracy. First, I argue that the policy initiative stalled because the debate was set up to depoliticize the question of health reform and thereby disengage citizens from the problem. Second, I defend a conception of "power sensitive" deliberation, which I argue is a check against two typical mechanisms of depoliticization: factionalism and the "democractic wish." Third, I suggest four criteria for power-sensitive deliberation, which I then use to analyze the recent health care debates. Finally, I explore the incentives and obstacles to creating opportunities for this kind of deliberation. PMID- 8708342 TI - Harry and Louise go to Washington: political advertising and health care reform. AB - Political advertising by interest groups trying to influence public policy has proliferated recently. Formerly the preserve of election campaigns, advertising has spread to policy arenas, such as abortion, trade, and health care. This article examines group lobbying for and against President Clinton's health care reform plan. Using a study of advertisements, a content analysis of news coverage, interviews with half a dozen leading figures in the debate during the spring of 1995, and an analysis of three national public opinion surveys designed to gauge the public response to health care ads, we investigate the media campaign on health care. Ads directed against the Clinton plan played a crucial role in the public's attaching negative connotations to some of its key elements. Grassroots campaigns can work either by mobilizing public opinion or by persuading political leaders that grassroots opposition exists to a particular program. PMID- 8708343 TI - Allocating health care: cost-utility analysis, informed democratic decision making, or the veil of ignorance? AB - Assuming that rationing health care is unavoidable, and that it requires moral reasoning, how should we allocate limited health care resources? This question is difficult because our pluralistic, liberal society has no consensus on a conception of distributive justice. In this article I focus on an alternative: Who shall decide how to ration health care, and how shall this be done to respect autonomy, pluralism, liberalism, and fairness? I explore three processes for making rationing decisions: cost-utility analysis, informed democratic decision making, and applications of the veil of ignorance. I evaluate these processes as examples of procedural justice, assuming that there is no outcome considered the most just. I use consent as a criterion to judge competing processes so that rationing decisions are, to some extent, self-imposed. I also examine the processes' feasibility in our current health care system. Cost-utility analysis does not meet criteria for actual or presumed consent, even if costs and health related utility could be measured perfectly. Existing structures of government cannot creditably assimilate the information required for sound rationing decisions, and grassroots efforts are not representative. Applications of the veil of ignorance are more useful for identifying principles relevant to health care rationing than for making concrete rationing decisions. I outline a process of decision making, specifically for health care, that relies on substantive, selected representation, respects pluralism, liberalism, and deliberative democracy, and could be implemented at the community or organizational level. PMID- 8708344 TI - Americans' political participation in the 1993-94 national health care reform debate. AB - The health politics and policy communities are still struggling with the question of "what went wrong" in the 1993-94 health care reform effort. Here I identify which Americans were politically active and inactive during the health care reform debate to explore the role political participation may have had in determining the outcome of the debate. Using data from a national and California random-sample telephone surveys, and controlling for other demographic attributes, I found that those who engaged in political activity specifically related to health care reform were disproportionately more likely to be self identified conservatives, less likely to favor an employer mandate plan, more likely to be fifty to sixty-four years old, more likely to be men, and more likely to have greater interest in and knowledge of the health care issue. Even in California, where a single-payer proposal was on the November ballot, self identified liberals were no more likely to engage in political activity on health care reform than were moderates or conservatives. I consider implications for the reform outcome given that liberals, the elderly, and those favoring the employer mandate proposal were all disproportionately "silent" during this debate, and finally I discuss the potential for mobilization during future debates. PMID- 8708345 TI - An exploratory study of the relationship between alternative therapies, functional status, and symptom severity among people with multiple sclerosis. AB - Roy's Adaptation Model guided an exploratory study of the association of alternative therapies with functional status and symptoms in people with multiple sclerosis (MS). Data were collected from 16 volunteers by a telephone administered interview schedule and a mailed questionnaire. All 16 respondents reported seeking traditional medical treatment and at least 1 alternative therapy (mode = 3, range = 1-9). The correlation between the number of alternative therapies and functional status was -.42; the greater the number of therapies used by the respondents, the lower their functional status. Qualitative data analysis revealed that functional status was enhanced by beneficial effects of specific alternative therapies, pacing oneself, changing the perception of self, and focusing on spiritual aspects of self. Self-reported MS symptoms were less severe following use of alternative therapies (p < .0005). Study of the differential effects of single and combined alternative therapies on functional status and symptom severity for large samples of people with MS is recommended. PMID- 8708346 TI - Modeling and role-modeling theory: a case study of holistic care. AB - To provide holistic care, nurses must attempt to view the world through the clients' eyes. The Theory of Modeling and Role-Modeling (Erickson, Tomlin, & Swain, 1983) enables nurses to care for and nurture each client with an awareness of and respect for the individual's uniqueness. This type of care giving exemplifies theory-based clinical practice that focuses on the clients' needs. Care is offered that recognizes that clients have the knowledge and ability to understand what has made them sick, as well as what will make them well. The authors explore the experiences of a young woman with diabetes mellitus and demonstrate how nursing care based on the Theory of Modeling and Role-Modeling enabled the client to recognize and develop strengths and empowered her to begin the healing process. PMID- 8708347 TI - Struggles with symptoms: women's narratives of managing HIV illness. AB - The purpose of this research report is to derive a holistic view of the symptom experiences of women living with HIV/AIDS. A racially diverse sample of 38 HIV infected women participated in in-depth narrative interviews describing their perceptions, evaluations, and responses related to HIV symptoms. Findings suggest a pattern to their struggles with symptoms: They experienced an accumulation of overwhelming symptoms, grew intensely fearful of these symptoms and what it meant to have HIV, and acted to protect themselves from allowing the HIV to "take hold". Many women perceived health care contact and cooperation with prescribed regimens as avenues for HIV to "take over" their lives, so they avoided these actions. Exemplar narratives illustrate analytic findings. PMID- 8708348 TI - The advancement of holistic nursing practice. PMID- 8708349 TI - Mental health nurses' spiritual perspectives. AB - Spiritual care has reemerged as a critical concern in nursing care. It is assumed that to provide spiritual care, nurses have a personal spiritual perspective. This study investigates the spiritual perspectives of a convenience sample of 50 mental health nurses employed in a public facility. The nurses' spiritual perspectives were measured using Reed's (1986a) Spiritual Perspective Scale (SPS). Due to the homogeneous nature of the sample and a clustering of SPS scores, the Wilcoxin Signed-Ranks Test was used to evaluate the data. Although no variables were identified as significant contributors to an individual nurse's spiritual perspective, an overall high SPS was found among the mental health nurses surveyed. The mental health nurses' SPS score mean (5.334, SD = .56) was notably higher than found i previous studies using the SPS (Reed, 1986a, 1991). PMID- 8708350 TI - Menstrual synchronization: a qualitative study. AB - Previous scientific studies documenting the objective existence of a coming together of the onset of menstrual cycles between or among women do not discuss the subjective meaning of the experience of menstrual synchronization from the perspective of women studied. This article discusses a phenomenologic study, based on 13 taped interviews and 2 written follow-up interviews, with women aged 25 to 46. Data were analyzed using a qualitative, naturalistic method along with a computer program. The analysis focused on the lived experience of menstrual synchronization. Four themes with respective subthemes emerged, producing descriptions of menstrual synchronization. The descriptions discovered can assist nurses to understand the holistic aspects of this everyday experience of women and to design effective strategies and techniques to help women gain knowledge about their cycle functions, promote healthy attitudes toward menstruation as a process, and acknowledge and honor this natural, healthy aspect of their menstrual cycle. PMID- 8708351 TI - Understaffing: a risk factor for infection in the era of downsizing? PMID- 8708352 TI - The role of understaffing in central venous catheter-associated bloodstream infections. AB - OBJECTIVE: To determine risk factors for central venous catheter-associated bloodstream infections (CVC-BSI) during a protracted outbreak. DESIGN: Case control and cohort studies of surgical intensive care unit (SICU) patients. SETTING: A university-affiliated Veterans Affairs medical center. PATIENTS: Case control study: all patients who developed a CVC-BSI during the outbreak period (January 1992 through September 1993) and randomly selected controls. Cohort study: all SICU patients during the study period (January 1991 through September 1993). MEASUREMENTS: CVC-BSI or site infection rates, SICU patient clinical data, and average monthly SICU patient-to-nurse ratio. RESULTS: When analyzed by hospital location and site, only CVC-BSI in the SICU had increased significantly in the outbreak period compared to the previous year (January 1991 through December 1991: pre-outbreak period). In SICU patients, CVC-BSI were associated with receipt of total parenteral nutrition [TPN]; odds ratio, 16; 95% confidence interval, 4 to 73). When we controlled for TPN use, CVC-BSI were associated with increasing severity of illness and days on assisted ventilation. SICU patients in the outbreak period had shorter SICU and hospital stays, were younger, and had similar mortality rates, but received more TPN compared with patients in the pre outbreak period. Furthermore, the patient-to-nurse ratio significantly increased in the outbreak compared with the pre-outbreak period. When we controlled for TPN use, assisted ventilation, and the period of hospitalization, the patient-to nurse ratio was an independent risk factor for CVC-BSI in SICU patients. CONCLUSIONS: Nursing staff reductions below a critical level, during a period of increased TPN use, may have contributed to the increase in CVC-BSI in the SICU by making adequate catheter care difficult. During healthcare reforms and hospital downsizing, the effect of staffing reductions on patient outcome (i.e., nosocomial infection) needs to be critically assessed. PMID- 8708353 TI - Tuberculosis in the workplace: OSHA's compliance experience. AB - OBJECTIVE: Inspections of 272 facilities were performed between May 1992 and October 1994 to determine compliance with applicable Occupational Safety and Health Administration (OSHA) requirements for prevention of tuberculosis (TB) transmission. DESIGN: Retrospective record review of two data sources: (1) OSHA's Computerized Integrated Management Information System and (2) an inspector completed questionnaire on inspection results. SETTING/PARTICIPANTS: Inspections of five types of facilities: healthcare institutions, correctional facilities, homeless shelters, long-term-care facilities for the elderly, and others, including drug treatment centers that the Centers for Disease Control and Prevention (CDC) identified as having a higher than expected rate of TB. METHODS: The OSHA Compliance Memorandum, based on the 1990 CDC Guidelines, which outlined elements of a TB prevention program, was used in performing 272 inspections of facilities between May 1992 and October 1994. Elements of compliance were recorded and reviewed from the IMIS database and inspectors' questionnaires. RESULTS: Regulated facilities were not fully compliant with OSHA guidance. Generally, healthcare facilities performed better than other facilities. Most facilities (79%) were compliant with administrative elements of a comprehensive TB control program, such as early identification of known or suspected infectious TB patients and skin testing of workers. Only 29% of inspected facilities were found to have acceptable respiratory protection programs for the prevention of occupational TB. CONCLUSION: Facilities have not been fully compliant with the OSHA memorandum describing protection of workers from TB. Facility compliance was better with some traditionally recognized TB infection control elements, but was weaker in the area of respiratory protection programs. This may reflect a lack of familiarity with the latter type of hazard protection. PMID- 8708354 TI - Staphylococcus aureus bacteremia: factors predicting hospital mortality. AB - Among 89 episodes of Staphylococcus aureus bacteremia, factors identified as independent predictors of hospital mortality were Acute Physiology and Chronic Health Evaluation III score > 60 (odds ratio [OR], 3.2; 95% confidence interval [CI95], 1.7 to 5.9) and Lifestyle score > 1 (OR, 2.1; CI95, 1.2 to 3.6). Future studies of S aureus bacteremia should take into consideration acute severity of illness (as well as treatment and source of infection) when evaluating outcome. PMID- 8708355 TI - Knowledge of the transmission of tuberculosis and infection control measures for tuberculosis among healthcare workers. AB - Two hundred seventy-five of 325 (85%) healthcare workers (HCWs) completed a questionnaire testing their knowledge of transmission of tuberculosis (TB) and infection control measures for TB. Of the 75 HCWs with no patient contact, 49 (65%) felt that masks should be worn, and 40 (53%) would use gowns. In contrast, 175 (88%) of 200 HCWs having contact with patients thought masks should be worn, and 70 (35%) would use gowns (P = .0001 and .0085, respectively). Only 87% of HCWs with patient contact felt that respiratory precautions should be instituted for TB patients. The results of our survey showed that HCWs should be reeducated about TB. PMID- 8708356 TI - An outbreak of cutaneous aspergillosis in a tertiary-care hospital. AB - A cluster of four cases of surgical and burn wound aspergillosis occurred in a 900-bed, adult, tertiary-care hospital. The source was traced to the outside packages of dressing supplies, which had become contaminated during construction in the central Inventory Control area. This resulted in patients with large exposed surface areas being inoculated directly with Aspergillus spores. PMID- 8708357 TI - Blood and body fluid exposures among surgeons: a survey of attitudes and perceptions five years following Universal Precautions. AB - A mail survey of surgeons at our medical center 5 years following adoption of Universal Precautions revealed that 29% estimated having > or = 1 potentially serious blood or body fluid exposures (BBEs) per month. Failure to report BBEs (usually needlesticks) to the employee health department was common, and the majority of surgeons in practice for > or = 10 years never reported such exposures. The most commonly cited reason for not reporting BBEs was perceived low risk of acquiring bloodborne infections. PMID- 8708358 TI - Unrecognized HIV-1 infection in inner-city hospital emergency department patients. AB - To determine the prevalence of unrecognized human immunodeficiency virus (HIV)-1 infections in patients presenting to an inner-city hospital emergency department, medical records were reviewed from 1,945 patients diagnosed with diseases not related to HIV or acquired immunodeficiency syndrome. The overall seroprevalence was 2.1% (40): 1.8% (11) in nontrauma versus 3.0% (29) in trauma patients. The highest prevalence was found in black, male, uninsured patients. PMID- 8708359 TI - A cluster of Legionnaires' disease in a community hospital--a clue to a larger epidemic. AB - Four patients with atypical pneumonias admitted within a 1-week period to a community hospital were suspected and confirmed to have Legionnaires' disease. The patients had a common travel history. This cluster of cases uncovered a larger epidemic associated with cruise travel. PMID- 8708360 TI - Comparative killing kinetics of methicillin-resistant Staphylococcus aureus by bacitracin or mupirocin. AB - The in vitro activities of bacitracin and mupirocin were compared for seven different strains of methicillin-resistant Staphylococcus aureus. Six of seven strains showed bacitracin minimum inhibitory concentrations (MICs) of 0.5 to 1.0 units/mL, and all seven had mupirocin MICs of 0.5 to 2 micrograms/mL. Time-kill studies revealed 2.6- to 4.5-log reduction in 24 hours with strains susceptible to bacitracin (4 units/mL) and 0 to 2.2 reduction with mupirocin (16 micrograms/mL). Bacitracin should be considered further for in vivo studies because of enhanced bacteriocidal effect and lower cost. PMID- 8708361 TI - Clostridium difficile contamination of blood pressure cuffs: a call for a closer look at gloving practices in the era of universal precautions. AB - We report an outbreak of Clostridium difficile-associated diarrhea at our medical center following adoption of Universal Precautions. Environmental cultures revealed unexpected contamination of blood pressure cuffs at a rate similar to that for bedside commodes (10% and 11.5%, respectively). An observational survey revealed that healthcare workers in the patient care areas not infrequently failed to remove their potentially stool-contaminated gloves prior to touching clean surfaces, which might have contributed to contamination of blood pressure cuffs. PMID- 8708362 TI - High-level disinfection or "sterilization" of endoscopes? AB - Controversy has surrounded the use of liquid chemical germicides to reprocess medical instruments that are damaged by heat sterilization. A review of the literature was performed to assess and compare the efficacy of disinfection and sterilization processes. The results of this review demonstrate that high-level disinfection of thoroughly cleaned endoscopes is not associated with a higher infection rate than is "sterilization". While there may be a theoretical distinction between the highest level of disinfection and sterilization, thorough cleaning eliminates clinical differences between the two. High-level disinfection is quick, effective, inexpensive, and recommended whenever heat sterilization is not feasible. A low-temperature sterilization process should be considered only if it is comparable in cost to disinfection or if it offers demonstrated advantages without damaging the instrument. Neither disinfection nor sterilization is likely to be effective if the instrument is not cleaned thoroughly after use. PMID- 8708363 TI - Educational needs and opportunities for the hospital epidemiologist. AB - The role of the hospital epidemiologist has changed substantially over the last 30 years as medical care has become more complex. The hospital epidemiologist needs training in methods for surveillance, prevention, and control of nosocomial infections. The hospital epidemiologist also must know how to apply these methods to other areas, including the epidemiology of noninfectious adverse outcomes of medical care. Training in hospital epidemiology should be a defined part of every infectious disease fellowship training program. Ancillary and additional training is available from several sources. PMID- 8708364 TI - The role of colonization in the pathogenesis of nosocomial infections. AB - The close relationship between colonization and the development of nosocomial infections has been demonstrated. Patient-related factors, such as underlying illness for all major sites of infection and advanced age, and pathogen-related factors, such as the ability of bacteria to adhere to epithelial cells, play the major roles in the pathogenesis of colonization. However, exact mechanisms of colonization have not been elucidated, and modulation of bacterial adherence as a method of infection prevention remains experimental. Current methods of infection prevention, therefore, focus either on preventing growth of colonizing microorganisms or on preventing patient-pathogen contact. Topical antibiotics have been used as a method of colonization prevention. However, their effectiveness may be limited by increases in antibiotic resistance; moreover, the effects of patient outcome are controversial. Maintenance of the physiologic mucosal environment using nonantimicrobial agents seems a promising approach, but only a few studies demonstrating efficacy have been published. Prevention of colonization still must rely heavily on basic infection control measures to prevent contact between patient and pathogen. PMID- 8708365 TI - Abnormal phagocytic function in children under one year of age. AB - The action of white blood cells (WBC) against Proteus mirabilis was assayed in 22 children following meningitis or septicaemia and compared with that in control groups of: (1) adult volunteers, (2) children undergoing minor surgical procedures for non-infective conditions, and (3) children being investigated for suspected renal disease. WBC bacterial killing tests were normal in all 55 adult volunteers. Abnormal results were obtained in 11 of 22 (50%) children with meningitis or septicaemia, 10 of 21 (47.6%) children undergoing surgery and six of 13 (46%) children with suspected renal disease, thereby indicating that there were no significant differences in phagocytic function among these three groups. Of children aged 1 year or less, 75% had abnormal phagocytic function. The significance of this finding is discussed. PMID- 8708366 TI - Comparison of the polymerase chain reaction and serology for the diagnosis of HTLV-I infection. AB - Human T-lymphotropic virus type I (HTLV-I) is associated with tropical spastic paraparesis (TSP) and adult T-cell leukaemia/ lymphoma (ATL). HTLV-I seroconversion may not take place for many years after infection and some patients develop relatively low concentrations of antibodies that are difficult to detect by means of conventional assays. Some serologic tests also yield high rates of false-positive results. We therefore decided to determine whether the polymerase chain reaction (PCR) might be more sensitive than serology for the diagnosis of HTLV-I infection. Samples of serum and of peripheral blood mononuclear cells were obtained from 50 patients with spastic myelopathy and nine with T-cell neoplasia. The serum samples of serum were tested for the presence of HTLV-I antibodies by means of an enzyme-linked immunosorbent assay (ELISA). Positive results were confirmed by Western blot (WB) assay. DNA was extracted from the peripheral blood mononuclear cells and a PCR performed by use of two primer pairs from the env and pol regions of the proviral genome. Seven of the 50 patients with myelopathy and none of those with T-cell neoplasia had detectable antibodies to HTLV-I and all seven were PCR positive. One patient with an indeterminate WB result and one who was HTLV-I antibody-negative, both with myelopathy, were also PCR-positive. In this study, PCR was found to be more sensitive than serology for the diagnosis of HTLV-I infection. PCR should therefore be considered for selected HTLV-I antibody-negative patients with unexplained spastic myelopathy or T-cell neoplasia. PMID- 8708367 TI - Polymerase chain reaction for rapid diagnosis of respiratory adenovirus infection. AB - Endemic (type 1, 2, 5 and 6) and epidemic (type 3, 4 and 7) respiratory adenovirus infections are associated with upper respiratory tract symptoms, pharyngoconjunctival fever, and pneumonia. Improved methods of diagnosis are needed, particularly in immunocompromized patients. We examined 93 throat swabs or nasopharyngeal aspirates from patients with acute respiratory disease using virus isolation and an adenovirus-specific polymerase chain reaction (PCR) based on consensus primers H1 and H2 derived from the hexon region DNA sequences of serotypes 2 and 5. Specimens which yielded viruses other than adenovirus in cell culture (n = 23) or which were negative for infectious viruses (n = 25) were negative in the PCR. The sensitivity of DNA amplification was 76% (34/45) in comparison with virus culture, being markedly lower with subgenus B (types 3 and 7) strains than with subgenus C (type 1, 2, 5 and 6) isolates (8/16 (50%)) vs. 26/28 (93%). P = 0.004) despite the use of a low annealing temperature to maximize detection of adenoviruses belonging to subgenera other than C. Of the 11 samples falsely negative in a single-round PCR but yielding adenovirus type 1 (n = 1), type 2 (n = 1). type 3 (n = 7), type 7 (n = 1), or untyped isolates (n = 1) in cell culture, nine (82%) gave positive results after nested DNA amplification. Possible approaches to further improving the performance of adenovirus PCR with respiratory specimens are discussed. PMID- 8708368 TI - Adverse reactions to Japanese encephalitis vaccine in travellers. AB - Vaccination against Japanese Encephalitis (JE) has been carried out extensively in many Asian countries for the past 20 years. The vaccine was generally considered to be effective and of low reactogenity. However, since 1989 an unusual number of systemic reactions characterized mainly by generalized urticaria and/or angioedema following JE vaccination were reported from Australia, Canada and Denmark, 860 travellers were recruited during a period of 16 months for a prospective study with the aim to investigate the type and incidence of side effects following JE vaccination (JEV) in German travellers. 826 received a primary immunization (2 injections at days 0 and 7-14) and 34 received a single booster injection. A detailed standardized questionnaire was distributed to all vaccinees after the first injection. A total of 509 questionnaires could be evaluated, which represents a return rate of 59.2%. 46% of the vaccinees reported about no adverse events at all. 54% reported about one or more adverse effects. Local reactions at the injection site were observed by 209 vaccinees, while 65 reported about systemic side effects like headache, fever, dizziness and generalized rash. There was no significant difference following first or second injection of the primary immunization or the booster injection, respectively, regarding incidence, severity or type of side effects. 2.2% of the vaccinees reporting reactions sought medical advice and 1.8% were judged unfit for work for an average of 2.2 days. The amount of systemic reactions might indicate a potential hazard of serious anaphylactic reactions. Unlike hepatitis A. Japanese encephalitis is an extremely rare disease in travellers. Therefore, the risk of acquiring the disease when travelling to affected areas without prior immunization should be considered against the risk of developing serious side effects after vaccination. We conclude that JEV should remain restricted to travellers with an increased risk of acquiring JE. PMID- 8708369 TI - Risks and benefits of aerosolized pentamidine and cotrimoxazole in primary prophylaxis of Pneumocystis carinii pneumonia in HIV-1-infected patients: a two year Italian multicentric randomized controlled trial. The Italian PCP Study Group. AB - We randomized 220 HIV-1-infected subjects to receive aerosolized pentamidine (300 mg/4 weeks) or orally trimethoprim-sulfamethoxazole (320-1600 mg/day) for primary prophylaxis of Pneumocystis carinii pneumonia (PCP), and evaluated PCP and toxoplasmic encephalitis (TE) occurrence and survival. Patients developing toxicity switched to the other regimen. Analysis was on intention-to-treat. At 1 year of study, we observed in the pentamidine group a non-significant excess of PCP (4 vs. 1) and TE (7 vs. 3), and a significant increased death rate (15 vs. 2). After 2 years, no significant differences were observed: adjusted RR estimates for pentamidine vs. cotrimoxazole were 1.20 (95% CI, 0.33-4.37) for PCP (6 cases vs. 5), 1.23 (95% CI, 0.46-3.29) for TE (10 vs. 8) and 1.52 (95% CI, 0.83-2.79) for death (30 vs. 18). Crossovers were more frequent in the cotrimoxazole group (41 vs. 4, P < 0.001). Aerosolized pentamidine and cotrimoxazole were equally effective in preventing PCP, and no major differences were observed in TE occurrence and survival after 2 years follow-up. PMID- 8708370 TI - Visceral leishmaniasis in HIV infected patients: treatment with high dose liposomal amphotericin B (AmBisome). AB - Visceral leishmaniasis (VL) in patients coinfected with human immunodeficiency virus (HIV) is often atypical, and characteristically relapses after treatment. We treated 10 HIV infected patients (9 men) with parasitologically confirmed VL with liposomal amphotericin B ("AmBisome': L-AMB) at a dose of 4 mg/kg/day on days 1, 2, 3, 4, 5, 10, 17, 24, 31, and 38. Patients were hospitalized for the first 5 days, and were monitored during, and 1 week and 1, 3 and 6 months after, L-AMB therapy. There were no serious adverse events, and L-AMB was well tolerated. 9/10 patients completed therapy, one patient defaulted at day 24. Clinical improvement was seen in all nine patients and the bone marrow aspirate was cleared of visible/culturable parasites in 8/9 patients. During follow-up, one patient defaulted. The seven remaining patients relapsed at 2, 3, 3, 5, 5, 6 and 7 months. Re-treatment with a variety of antileishmanial drugs was unsatisfactory. The time from first diagnosis of VL to death in six patients was 5-40 months (mean 18.8 months). Only one patient remained alive 26 months after treatment. L-AMB is safe and provides a good initial clinical response. Intermittent dosing enables a short period of hospitalization. However, relapse is probably inevitable. PMID- 8708371 TI - Serotypes and antibiotic susceptibilities of Streptococcus pneumoniae in Nairobi, Kenya. AB - Two hundred and sixteen clinically significant isolates of Streptococcus pneumoniae from 138 adult patients attending clinics in Nairobi, Kenya over a 2 year period were characterized by antibiotic sensitivity testing and serotyping. Overall antibiotic resistance rates were: penicillin, 25%; tetracycline, 34%; erythromycin, 0%; chloramphenicol, 0.4%. Minimum inhibitory concentrations (MIC's) of penicillin ranged from < 0.008-0.5 microgram/ml. Determination of penicillin resistance (MIC > or = 0.1 microgram/ml) by oxacillin 1 microgram disc diffusion zone diameter < or = 20 mm was 100% sensitive, 92% specific. Relative resistance to oxacillin (MIC range 0.25-1.0 microgram/ml) accounted for penicillin sensitive isolates determined falsely to be penicillin resistant by oxacillin disc testing. Penicillin resistance was more frequent in sputum isolates at 35% than in blood isolates at 18% (P = 0.013). Serotypes 6, 10, 14, 16, 19 and 23 were associated with penicillin resistance. This study provides information of value for planning management strategies for pneumococcal disease from an area where there are few existing data. PMID- 8708372 TI - Septic arthritis caused by an unusual type of Haemophilus influenzae. AB - A healthy 3.5-month-old infant developed septic arthritis with Haemophilus influenzae type f isolated from the knee aspirate and blood. The patient had no obvious risk factors and immunological evaluation revealed no abnormalities. To our knowledge, there are only two reported childhood cases of septic arthritis caused by non-type b H. influenzae, but this is the first one to be reported in a child without an underlying disorder. The importance of serotyping H. influenzae isolates in the H. influenzae type b vaccine era, and the need to look for predisposing factors in paediatric patients with invasive disease caused by non type b H. influenzae are discussed. PMID- 8708373 TI - Schizophyllum commune: an unusual isolate from a patient with allergic fungal sinusitis. AB - We describe the first case of allergic fungal sinusitis (AFS) associated with Schizophyllum commune. Histological diagnosis was made on the mucinous material from the sinus which contained eosinophils, fungal hyphae and Charcot-Leyden crystals. The fungal isolate was identified as S. commune on the basis of its colonial morphology and minute peg-like outgrowths from vegetative hyphae and clamp connections. The optimal treatment of AFS is not known but may involve surgical debridement followed by systemic corticosteroids, gradually reduced, with the patient being maintained on inhaled corticosteroids. Oral itraconazole could be added to the regimen of those patients in whom frequent recurrences occur after debridement or when there is histological evidence of severe pressure erosion. PMID- 8708374 TI - Acute transverse myelitis associated with Chlamydia psittaci infection. AB - We report a case of transverse myelitis presenting with fever and rapidly deteriorating neurological signs associated with Chlamydia psittaci infection. It resolved with no long-standing neurological consequences. Only one previous report has described transverse myelitis in association with psittacosis, and this resulted in permanent neurological sequelae. PMID- 8708375 TI - Multidrug-resistant tuberculosis in an HIV-positive man in the United Kingdom. AB - Multidrug-resistant Mycobacterium tuberculosis infection (MDR-TB) in those who are HIV positive has until now been largely a North American phenomenon. We report a fatal case in London. PMID- 8708376 TI - Quinolone resistance in Oligella urethralis-associated chronic ambulatory peritoneal dialysis peritonitis. AB - Oligella urethralis is an organism which is normally isolated as a commensal from the genitourinary tract. We describe the first two reported cases of CAPD associated peritonitis caused by this organism. Both isolates were found to be resistant to ciprofloxacin, while relatively sensitive to a wide range of antimicrobial drugs. These findings indicate that this organism may be an opportunistic pathogen for CAPD patients, and that extensive ciprofloxacin usage provides a selection pressure for emergence of resistance. PMID- 8708377 TI - Tuberculous pericarditis in a child. AB - A 14-year-old boy with tuberculous pericarditis and tamponade is described. Confirmation was by culture of pericardial aspirate. Though he did not have a cough, acid-fast bacilli were detected following induced sputum. Chest X-ray did not show evidence of pulmonary tuberculosis, but enlarged mediastinal nodes were detected by computerized tomography. He made an excellent response to anti tuberculous chemotherapy and corticosteroids. PMID- 8708378 TI - Escherichia coli infection of an abdominal aortic aneurysm. PMID- 8708379 TI - Lactobacillus rhamnosus infection in a child following bone marrow transplant. AB - We report a case where Lactobacillus rhamnosus was isolated from pericardial effusion and blood in a child following a bone marrow transplant for aplastic anaemia. A resume of cases in which this organism has been implicated as a pathogen is also presented. PMID- 8708380 TI - Molecular epidemiology of human rotaviruses: reassortment in vivo as a mechanism for strain diversity? PMID- 8708381 TI - Sub-acute intestinal obstruction by Strongyloides stercoralis. PMID- 8708382 TI - Post Salmonella irritable bowel syndrome--5 year review. PMID- 8708383 TI - Cholera in mainland Europe probably associated with the consumption of imported coconut milk. PMID- 8708384 TI - Abuse of Brolene eye drops with putative corneal infection. PMID- 8708385 TI - Mixed cerebral pathology in AIDS patients. PMID- 8708386 TI - Diagnosis of chronic strongyloidiasis. PMID- 8708387 TI - Neisserial disease. PMID- 8708388 TI - Clinical presentation, diagnosis and therapy of progressive multifocal leukoencephalopathy. PMID- 8708390 TI - Analysis of small cystic lesions of the pancreas. AB - There have been few reports on (1) the nature and pathogenesis of small cystic lesions of the pancreas, (2) their incidence, age distribution, and location, and (3) their significance as potential precursors of intraductal papillary tumors, mucinous cystic tumors, and duct cell carcinomas. MATERIALS: Epithelial growth of small cystic lesions in 300 consecutive autopsy cases and in seven cases of small duct cell carcinoma from among 2300 elderly autopsy cases, was evaluated by histopathological analysis. One hundred eighty-six cystic lesions were found in 73 of 300 autopsy cases (24.3%). The incidence of cystic lesions increased with age. Cystic lesions were equally distributed throughout the pancreas. Epithelial atypia was histologically classified into five groups: normal epithelium; papillary hyperplasia without atypia; atypical hyperplasia; carcinoma in situ; and invasive carcinoma. The incidence of each group was 47.5, 32.8, 16.4, 3.4, and 0%, respectively. Epithelia of atypical hyperplasia or carcinoma in situ were more prevalent in small cystic lesions (less than 4 mm in diameter) than in larger lesions (chi-square test, p < 0.05). Epithelia of dilated ductular branches adjacent to cystic lesions showed a similar degree of atypia as the epithelia of the cystic lesions themselves (p < 0.01). Epithelial atypia of the main pancreatic duct was mild in all of the cases but two, and was not related to that of the cystic lesion. Among the seven cases of small duct cell carcinoma, two cases had small cancerous cystic lesions, 4.1 and 5.3 mm in diameter, within the tumor. Small cystic lesions appear to have the potential to progress to malignancy but definitive evidence has not been demonstrated. Additional studies, including molecular biological examinations, are necessary to fully understand the biology of these lesions. PMID- 8708391 TI - Fine structure of the cilia in the pancreatic duct of WBN/Kob rat. AB - We observed the cilia in pancreatic ducts (intraductal cilia) with scanning and transmission electron microscopy (SEM and TEM), using male WBN/Kob rats (W/K), which are the spontaneously developed chronic pancreatitis models with stasis of pancreatic juice, and male Wistar rats as control. By SEM observations, the lengths of cilia in interlobular ducts of 18-mo-old W/K were demonstrated to elongate markedly. By TEM observations in the controls, cross-sections of the intraductal cilia were demonstrated to present various numbers of microtubules (those with seven, eight, or nine microtubules accounted for 83.3% of all). There was no significant difference between W/K and controls in the number of microtubules in the cross-sections of intraductal cilia: the intraductal cilium core was provided with nine microtubules, which were different from the number of microtubules encountered within the cilium core of other ciliated cell (i.e., bronchial epithelium, and so forth), and their number in the cross-section of intraductal cilium decreased at a distal portion. Though some of their cross sections revealed deformities of ciliary membranes and disarrangements of microtubular complex, there was no significant difference in their incidence between both rats. These findings suggest that the intraductal cilia have different functions from the ciliated cells' cilia, and W/K has the elongated intraductal cilia without internal structural change. PMID- 8708389 TI - Neural regulation of the endocrine pancreas. PMID- 8708392 TI - Detection of point mutations in K-ras gene at codon 12 in bile from percutaneous transhepatic choledochal drainage tubes for diagnosis of biliary strictures. AB - Detection of K-ras mutations at codon 12 constitutes one modality for diagnosis of pancreatic tumors. We attempted to detect K-ras mutations in DNA from bile collected through percutaneous transhepatic choledochal drainage (PTCD) tubes as a diagnostic approach to biliary strictures. Since bile salts induce cell damage, we first investigated the degeneration of cells according to bile exposure time using cell lines. High-mol-wt DNA could be extracted from cells exposed to bile for 6 h, but not from those exposed for 12 h. However, DNA exposed to bile for up 12 h could be amplified by the polymerase chain reaction (PCR) method. Therefore, K-ras mutations in fresh bile specimens collected from 15 patients through PTCD tubes were examined using PCR with restriction enzyme digestion. K-ras mutations were found in five out of five (100%) pancreatic cancers, all of which were negative according to cytodiagnosis of the same bile. On the other hand, K-ras mutations were not detected in bile from biliary tract cancers or metastatic neoplasms, except for one bile duct carcinoma and one metastatic case. Thus, although K-ras mutation alone is not an absolute marker for cancer, detection of K-ras mutations in fresh bile from PTCD tubes is a useful adjunct for diagnosis of pancreatic carcinomas in cases of biliary tract strictures. PMID- 8708393 TI - Elevated serum levels of immunoreactive anionic trypsin (but not cationic trypsin) signals pancreatic disease. AB - Pancreatic juice from most studied species contains two major forms of trypsin, one with anionic electrophoretic mobility and one with cationic mobility. They are referred to as anionic and cationic trypsin(ogen). The purpose of this study was to measure immunoreactive anionic trypsin (irAT) and immunoreactive cationic trypsin (irCT) in sera from patients with pancreatic cancer (n = 39) and chronic pancreatitis (n = 32) using two specific ELISA methods. Sera from 72 healthy persons were used as controls. Patients with pancreatic cancer showed significantly elevated serum levels of irAT median level 39 vs 20.5 micrograms/L in the control group (p < 0.001). No differences in irCT levels were found. The ratio between irAT and irCT in serum was significantly increased (p < 0.001). Patients with chronic pancreatitis showed a wide range of both irAT and irCT levels, but no significant differences compared to the control group. The ratio between irAT and irCT was, however, significantly increased also in this group of patients. The results suggest a nonparallel secretion of anionic and cationic trypsinogen in pancreatic disease. This is a pattern that has been observed in experimental forms of chronic "hyperCCKemia." PMID- 8708394 TI - Altered expression of extracellular matrix molecules and their receptors in chronic pancreatitis and pancreatic adenocarcinoma in comparison with normal pancreas. AB - The aim of this study was to elucidate the expression and distribution patterns of both integrins and extracellular matrix (ECM) molecules in chronic pancreatitis (CP) and pancreatic adenocarcinoma (PC) compared with normal pancreas (NP). Expression of nine alpha-subunits (alpha 2-alpha 6, alpha V, alpha L, alpha M, and alpha X), four beta-subunits (beta 1, beta 3-beta 5), and four ECM molecules (type IV collagen, laminin, fibronectin, and vitronectin) was investigated immunohistochemically. In CP, all integrins except alpha V showed nearly the same staining patterns compared with NP. Some acinar cells in CP expressed alpha V. Whereas alpha 2, alpha 3, and alpha 6 expression was stronger and diffuse, no alpha 5 expression was seen in PC. Basement membrane (BM) showed continuous staining in CP, whereas it showed discontinuous/absent staining in PC with antitype IV collagen, laminin, and vitronectin antibodies. Some carcinoma cells showed reverse correlation between alpha 2, alpha 3, and alpha 6 expression and type IV collagen and laminin expression. Fibronectin showed diffuse stromal expression in CP and PC. Some acinar cells or duct cells in CP carcinoma cells in PC showed intracellular VN expression. These results suggest that these integrins and ECM molecules are involved in inflammatory and malignant processes in pancreas. PMID- 8708395 TI - Chronic pancreatitis and extrapancreatic cancer: a retrospective study among 181 patients with chronic pancreatitis. AB - The relationship between chronic pancreatitis (CP) and extrapancreatic cancer has been debated in the recent years. In prospective studies, it has been found that pancreatic cancer develops in 0-5% of patients with chronic pancreatitis. Many papers describe an increased relative risk for developing extrapancreatic cancer in patients suffering from chronic pancreatitis. In this study including 181 patients with CP, we found 14 patients with extrapancreatic cancer (three of these had two different types of cancer). No patient had pancreatic cancer. It was found that the respiratory airways and upper gastrointestinal tract were the dominating locations (five and four cases, respectively), but also genital and hemolymphopoietic cancers were represented (four and two cases, respectively). Two patients had metastatic cancer with unknown primary tumor. The patients with cancer tended to be older than those without cancer. The patients with CP had a 2.43 times greater risk of developing cancer than the general Danish population (age and sex standardized comparison). The relatively large number of cancers in the upper gastrointestinal tract and respiratory airways suggest that tobacco and alcohol may be responsible, as these organs have the highest exposure to these compounds, which are well known carcinogens. PMID- 8708397 TI - A novel adhesion factor produced by hamster pancreatic cancer cell line is effective on normal and carcinoma cell lines of different species. AB - An adhesion factor, produced by the hamster pancreatic cancer cell line PC-1.0, was tested for its efficiency in promoting the in vitro adhesion of normal and tumor cells (pancreas, lung, kidney, colon, breast, skin, prostate, neuroblast, melanocyte) derived from human, monkey, bovine, hamster, and rat sources. Using a modification of the dimethylthyazol diphenyl tetrazolium (MTT) assay, the factor was found to induce adhesion in all cell lines in a dose-dependent manner. Although the effect was variously expressed, there was a statistically significant difference between the MTT absorbance of cells incubated in the presence or absence of the factor. Conditioned medium of each cell line tested showed significantly less adhesion effect than that produced by PC-1.0 cells. Because our previous study indicated that the adhesion factor produced by PC-1.0 cells differed from known growth factors and adhesion molecules including fibronectin, vibronectin, laminin, and collagen, it appears that PC-1.0 cells produce a novel adhesion factor that enhances adherence of normal and malignant cells of different species. PMID- 8708396 TI - Combined use of nuclear morphometry and DNA ploidy as prognostic indicators in nonresectable adenocarcinoma of the pancreas. AB - The introduction of combined modality treatment has only marginally affected the prognosis in nonresectable pancreatic carcinoma. Evaluation of easily accessible prognostic variables could be of clinical importance when selecting patients for proper therapy. DNA ploidy and morphometric variables were chosen as prognostic markers and assessed on cytologic material obtained by fine-needle aspiration biopsy (FNAB) from 128 patients with pancreatic carcinoma. The nuclear DNA content was measured by image cytometry. Patients were categorized as short-term (< or = 6 mo) and long-term survivors (> 6 mo). Ninety-eight of 116 nonresectable patients were possible to evaluate. There were significant differences between short- and long-term survivors with regard to DNA ploidy (p < 0.01) and the morphonuclear variable anisokaryosis (p < 0.001). In patients with either DNA aneuploid tumors or anisokaryosis > or = 0.6, the survival time was 6 mo or less in 85 and 93%, respectively. When both these criteria were fulfilled, only 5% survived for more than 6 mo. Thus, DNA ploidy and morphometry, separately or in combination, may provide prognostic information in nonresectable pancreatic carcinoma. PMID- 8708398 TI - Structural and functional changes of rat exocrine pancreas exposed to nicotine. AB - Nicotine, an addictive agent in cigarette smoking, has been implicated in the etiology of pancreatitis and pancreatic carcinoma. Very little experimental data are, however, available regarding the effects of nicotine on the structural and functional changes in the exocrine pancreas. Two groups of rats divided into control and nicotine exposed were used. The animals in the nicotine group were maintained for 28 d with ad libitum water containing 0.77 mM of nicotine. At term, the animals were sacrificed, and pancreas was examined for pathological and functional changes. Nicotine induced cytoplasmic vacuolation and cellular edema in the exocrine pancreas. Plasma levels of glucose and insulin and CCK-8 stimulated amylase release in isolated acini were significantly decreased by nicotine, whereas the total cellular amylase content was significantly increased. Analysis of competitive ligand binding data on membranes from isolated acini showed that the Bmax and Kd values for CCK receptors were not significantly changed by nicotine, p > 0.05. These data indicate that a postreceptor mechanism is involved in the inhibition in stimulus-secretion coupling of enzyme secretion by nicotine. The increase in total cellular amylase content and decreased enzyme secretion by nicotine may be implicated in the induction of pancreatic pathology. PMID- 8708400 TI - Von Hippel-Lindau disease complicated by acute pancreatitis and Evan's syndrome. AB - Von Hippel-Lindau syndrome (VHL) is an autosomal dominant disorder characterized by renal cysts, retinal angiomas, central nervous system hemangioblastomas, and pancreatic cysts. Evan's syndrome is a hematologic disorder characterized by autoimmune thrombocytopenia and autoimmune hemolytic anemia. We present the first case of acute pancreatitis and Evan's syndrome that developed in a patient with von Hippel-Lindau syndrome. PMID- 8708399 TI - CT-guided aspiration of suspected pancreatic infection: bacteriology and clinical outcome. AB - We have performed CT-guided percutaneous needle aspiration in 104 patients with severe pancreatitis strongly suspected of harboring pancreatic infection on the basis of systemic toxicity and CT findings (Balthazar CT grade D or E). Of these 104 patients, 51 (49%) were documented with pancreatic infection. Gram stain was positive in 54 of 58 infected aspirates, and culture was positive in all 58. Klebsiella, Escherichia coli, and Staphylococcus aureus were the most frequent organisms. Eighty-six percent of infected processes contained only one organism. Overall, pancreatic infection was documented by GPA within the first 2 wk in approx one-half of patients. There were no complications. The overall rate of infection decreased from 60 (1980-1987) to 34% (1988-1995) (p = 0.011). This change was caused by a reduction in the rate of infected necrosis from 67 to 32% (p = 0.015). The overall mortality rate remained at 20%. The mortality of sterile pancreatitis was not different from infected pancreatitis (p = 0.14). We conclude that GPA is a safe, accurate method of diagnosis of pancreatic infection. The rate of pancreatic infection appears to be decreasing. The overall mortality of severe pancreatitis among patients suspected of harboring pancreatic infection has remained unchanged because of the high mortality associated with both infected necrosis and severe sterile necrosis. PMID- 8708401 TI - Inflammatory pseudotumor of the pancreas. AB - We describe a rare example of inflammatory pseudotumor of the pancreas in a 42-yr old woman, which developed following chemotherapy for lymphoma of the uterine cervix. The patient had developed fatigue, weight loss, abdominal pain, and anemia; abdominal CT scan showed a large mass in the pancreas. Examination of the resected specimen revealed a fleshy, well-circumscribed, 7-cm mass. Histologically, there was a hypocellular to moderately hypercellular, bland spindle-cell proliferation admixed with a prominent infiltrate of lymphocytes, histiocytes, and plasma cells. The spindle cells were vimentin positive but negative for muscle markers; electron microscopy revealed only fibroblastic cells. DNA analysis revealed a diploid population with low S-phase fraction. The patient was well at 6-mo follow-up. It is important for the pathologist to be aware of the existence of this entity in unusual locations such as the pancreas so as to avoid a mistaken diagnosis of malignancy. PMID- 8708402 TI - [Usefulness of endotoxin-specific limulus test for the measurement of endotoxin in cerebrospinal fluid in diagnosis of bacterial meningitis]. AB - Using a new endotoxin-specific chromogneic limulus assay (Endoscopy test), endotoxin concentrations were measured in 93 specimens of cerebrospinal fluid (CSF) from 66 pediatric patients. Eighteen patients were diagnosed as having menigitios. Of these, 6 cases (group A) with gram-negative meningitis proven by culture had high CSF endotoxin concentrations of 115.3, (82-133) (median, range) pg/ml. Ten cases (group B) with gram-positive or aseptic meningitis had endotoxin concentrations of 2.15 (0.1-3.6) pg ml. Other 2 cases with bacterial meningitis (group C), in whom no pathogen was detected, had CSF endotoxin concentrations of more than 100 pg/ml. Four cases with encephalitis (group D) and 45 cases with non meningitis or non- encephalitis (group E), had CSF endotoxin concentrations of less than 5 pg/ml. Despite a negative culture after antibiotic treatment in group A patients, endotoxin was cleared slowly from the CSF. A clearing of endotoxin from CSF was followed by alleviation of fever with a more gradual decline in CRP values. In 2 cases of group C, the negative bacterial culture appeared to be attributable to the previous treatment with antibiotics. However, these patients had high CSF endotoxin levels, indicating gram negative bacterial meningitis. In 17 CSF specimens from 5 patients of group A, in whom Haemophilus influenzae was detected on admission, an additional a latex agglutination test for the detection of H. influenzae polysaccharide antigen was performed. Only 3 specimens from 3 patients with CSF endotoxin concentrations of more than 80 pg/ml had a positive agglutination test. These results suggest that quantitation of endotoxin concentrations is useful for the diagnosis of gram-negative meningitis. And also, the clearance of endotoxin from CSF during treatment appears to be useful in determining the timing of when antibiotic should be stopped. PMID- 8708403 TI - [Effects of antiseptics against methicillin-resistant Staphylococcus aureus]. AB - Various antiseptics, commonly used in hospital, were tested against methicillin resistant Staphylococcus aureus (MRSA) at high concentrations (approximately 10(9) CFU/ml) and in short time exposures. The antiseptic solutions of 0.05% alkyldiaminoethylglycine hydrochloride, 0.05% benzalkonium chloride, 0.2% povidone-iodine gargle and 0.03% dominophen bromide produced 1,000 to 1/100,000 reduction after a 30 sec exposure. 7.5 % povidone-iodine scrub and 0.2% benzalkonium chloride in 83% ethanol were most effective, reducing MRSA under detection limits. (less than 10 CFU/ml) 0.05% chlorhexidine gluconate was less effective, producing no evident reduction even after 1 min of exposure. Though 0.2% povidone solution reduced MRSA under detection limits at 37, 24 hrs incubation, colonies of several strains were detected after 48 hrs extended incubation. This suggests the existence of strains resistant to the antiseptic solution of povidone-iodine. We recommend that the disinfection for these strains should be repeated short time disinfection, because repeated short time exposure (15 sec 2 times) of antiseptics was more effective than one long time exposure (30 sec once). PMID- 8708404 TI - [Epidemic of influenza in Kyushu-Okinawa District (April 1994-March 1995)]. AB - The prevalence of influenza in Kyushu-Okinawa District in April 1994- March 1995 was studied as the prevalence of influenza virus, to determine the sero-type of influenza viruses isolated in Kyushu- Okinawa District prefectures and cities. As a result, three sero-types of influenza viruses, i.e. type A/H1N1, type H3N2 and type B, were isolated in Kyushu-Okinawa District in this season, but most of the isolates were type A/H3N2 and type B. Weekly changes of reported influenza patients and period of virus isolation at local public health institutes revealed that influenza epidemics of the earlier part in this season was caused by type A/H3N2 and the latter part due to type B. Type A/H3N2 spread all over Kyushu Okinawa District in a shorter period (about 2 weeks) through the westside of Kyushu and down south, and type B stayed about one month in northern Kyushu and took about 7 weeks to spread all over Kyushu-Okinawa District. Based on these results, the spread of influenza virus in Kyushu-Okinawa District was visualized on the isopleth maps. PMID- 8708405 TI - [Pathogenic bacteria isolated from the sputum of the patients with pulmonary emphysema]. AB - Isolated pathogenic bacteria from sputum of the patients with pulmonary emphysema who were admitted in our hospital from 1984 to 1994 were examined to elucidate the relationship between isolated bacteria from sputum and pulmonary functions including vital capacity (VC), forced expiratory volume (FEV1.0), PaO2 and PaCO2. VC of the patients from whom MSSA (methicillin-sensitive Staphylococcus aureus) or Enterococcus faecalis (E. faecalis) were isolated was significantly lower than that of the patients from whom Streptococcus pneumoniae (S. pneumoniae), Branhamella catarrhalis (B. catarrhalis) or Haemophilus influenza (H. influenza) were isolated. FEV1.0 had a similar tendency as VC in terms of isolated organisms from the patients with emphysema. Similarly, PO2 of the patients from whom MSSA or E. cloacae were isolated was significantly lower than that of the patients from whom S. pneumoniae, B. catarrhalis or H. influenzae were isolated, and PCO2 of the patients from whom S. pneumoniae, B. catarrhalis or H. influenza were isolated. There was also impaired respiratory function in the patients from whom MSSA, Escherichia coli (E. coli), Pseudomonas aeruginosa (P. aeruginosa), Xanthomonas maltophilia (X. maltophilia) or Enterobacter cloacae (E. cloacae) were isolated, compared with those in the patients from whom S. pneumonia, B. catarrhalis or H. influenzae were isolated. These results suggest that isolated pathogenic bacteria are shifted from S. pneumoniae, B. catarrhalis or H. influenza to MSSA, E. coli, P. aeruginosa, X. maltophilia or E. cloacae in the course of impairment of respiratory function in pulmonary emphysema. The treatment and prophylaxis for acute exacerbation in pulmonary emphysema should be based on these results. PMID- 8708406 TI - [A clinical study of gastrointestinal flora in patients with, or without MRSA colonization in the upper-respiratory tract after introduction of preventive measures of hospital infection]. AB - In early 1980's methicillin-resistant Staphylococcus aureus (MRSA) was reported as a major pathogenic organism of geriatric hospital infection in Japan. At the same time in the A geriatric hospital MRSA infection was prevalent. To decrease nosocomial infections some active preventive measures against hospital infection were taken since Oct. 1991. After introduction of preventive measures of hospital infection in geriatric ward (190 beds) nosocomial bacteremia and pneumonia were markedly decreased in comparison to the episode number before introduction of prevention. However several patients with MRSA colonization were observed every month. The aim of this clinical study was to clear how frequent MRSA was isolated from the gastric juice and stool. Any MRSA was not observed in 63 cultured stool, but just one MRSA was isolated in patients with MRSA colonization. On the other hand gram-negative organisms, which were E. coli, P. aeruginosa, P. mirabilis etc., were frequently observed in cultured stool. In conclusion, we considered frequency of MRSA colonization in gastrointestinal space was not so high but rather very low. PMID- 8708408 TI - [Clinical utility of DNA fingerprinting by arbitrarily-primed polymerase chain reaction (AP-PCR) in nosocomial infection caused by methicillin-resistant Staphylococcus aureus]. AB - Recently, nosocomial outbreaks of MRSA have become an serious social problem in Japan. To examine the routes of transmission of MRSA, the establishment of an accurate MRSA typing system is essential. Previously, we reported that the DNA fingerprinting by AP-PCR might be a useful method to differentiate MRSA strains. In this study, we tried to investigate the clinical usefulness of DNA fingerprinting by AP-PCR using clinically-isolated MRSA strains. Twenty-four MRSA strains (12 with coagulase type IV, and 12 with coagulase type II) isolated from patients in our department were used. Other typing methods (the sensitivity of antibiotics, pulsed-field gel electrophoresis, and plasmid analysis) were also performed. As a result, the typing pattern by AP-PCR correlated well with other typing methods. MRSA strains with coagulase type IV showed almost the same pattern, suggesting that these strains were nosocomially transmitted. On the other hand, MRSA strains with coagulase type II showed various patterns, suggesting these strains were not nosocomially transmitted. In conclusion, the typing by AP-PCR seemed to be a useful tool evaluate a nosocomial MRSA transmission. PMID- 8708407 TI - [Study on purple pigmentation in five cases with purple urine bag syndrome]. AB - We reported five patients with purple urine bag syndrome (PUBS). Four patients had indicanuria, and, in three of them, purple pigmentation was reproduced by inoculating urinary isolates, in the broth with indoxyl sulfate. Klebsiella pneumoniae, Pseudomonas aeruginosa and Enterococcus avium were considered to produce the purple pigment in three patients. However, attempts to reproduce the purple pigment failed in two patients, and one of them did not have indicanuria. These results suggest that indicanuria is not necessarily required for the development of PUBS. PMID- 8708409 TI - Predominance of codon 215 mutation in reverse transcriptase-coding region of 3' azido-3'-deoxythymidine (AZT)-resistant HIV-1 isolates after long-term AZT therapy. AB - Among human immunodeficiency type 1 viruses (HIV-1) isolated during long-term 3' azido-3'-deoxythymidine (AZT) therapy, 4 condoms (Asp67, Lys70, Thr215, and Lys219) in the HIV-1-reverse transcriptase (RT)-coding region have been considered to be related to the AZT resistance of HIV-1. Therefore we determined these mutation patterns in HIV-1 isolates from patients undergoing long-term AZT treatment. In 41 clones of HIV-1 from 7 patients, the Thr215 mutation was the most predominant (97.6%), and more frequent than Asp67 (48.8%), Lys70 (31.7%) and Lys219 (9.8%) mutations. All 22 clones from isolates cultured in the presence of 1 microM AZT Thr215 mutation; such a high frequency was not found for the other 3 codon mutations. In a clinical follow-up study, Thr215 mutation appeared in the late stage of AZT treatment parallel with the emergence of AZT insusceptibility. It is worth noting that this Thr215 mutation to Tyr or Phe is a 2-nucleotide mutation in contrast to the 1-nucleotide mutations seen in the other 3 codons. Isolates with the single amino acid change of Thr215 of the RT-coding region obtained after long-term AZT treatment grew in the presence of 1 microM AZT. This single amino acid mutation in the Thr215 codon is the most important factor in AZT resistance. PMID- 8708410 TI - [Some characteristics of verotoxin-producing Escherichia coli strains isolated from sporadic diarrhea in Akita Prefecture]. AB - Six sporadic cases of VTEC infection have been confirmed in Akita Prefecture from June. 1991 to Nov. 1994. Six VTEC strains isolated in these cases were examined for their serotype, Vero toxin type, existence of eae gene and 60 MDA plasmid, and CVD probe reactivity. Of the 6 isolates, 5 were O157:H7. Two isolates of which possessed TV-1 and VT-2 genes and the rest of 3 possessed VT-1 and VT-2vh genes, VT-2 gene, respectively. One isolate was O26:NM possessing VT-1 gene. A primer pair, designated as EA-1 and EA-2, was designed for detecting eae gene in VTEC. Examination of the 6 VTEC isolates by PCR using EA-1 and EA-2 primers indicated that all of the isolates were eae positive. This was further confirmed by dot blot hybridization using a eae probe. All of the 6 isolates also harbored approximately 60 MDa plasmid, which hybridized with the CVD419 probe in southern blot analysis. These results supported the hypothesis that eae gene and the 60 MDa plasmid might play a role in the mechanism for VTEC to cause diseases including diarrhea, hemorrhagic colitis and hemolytic uremic syndrome in humans. Diversity observed in plasmid profile and VT type of the 6 isolates suggested that they were originated from independent sources, which could not be identified. PMID- 8708411 TI - [Serovars and drug susceptibility of Salmonella isolated from patients with sporadic diarrhea in Yamanashi Prefecture during the last decade (1985-1994)]. AB - A total of 1242 strains of Salmonella isolated from patients with sporadic diarrhea during the period from April 1985 to December 1994 in Yamanashi Prefecture, were tested for their serovars and drug susceptibility. The results can be summerized as follows: 1) The isolates were serologically classified into 54 different serovars. The predominant serovars were S. Enteritidis (39.6%), S. Typhimurium (16.6%), S. Oranienburg (4.8%), S. Hadar (4.6%), S. Litchfield (4.4%). 2) Serovars of S. Stanleyville, S. Hartford, S. Rissen, S. Chincol, S. Gaminara, S. Poona and S. Hvittingfoss were isolated from human sources for the first time in Yamanashi Prefecture. 3) Yearly frequency of isolates was 19.2% in 1989, 15.5% in 1994, 11.8% in 1993. 4) Monthly frequency of isolates was 21.2% in August, 17.1% in September, 12.0% in October, 11.2% in July. 5) The predominant age of isolates were 2 years of age (99 strains: 8%), 1 year of age (92 strains: 7.5%), 4 years of age (68 strains: 5.5%), 3 years of age (64 strains: 5.2%) and under 1 year of age (62 strains: 5.0%). 6) The rate of isolates from males was higher than females (Male: 58.0%, Female: 42,0%). 7) The frequency of resistant isolates was 62.5% in the period. The most predominant resistance pattern was SM single resistance because of the increase of S. Enteritidis. 8) The number of resistant strains of S. Enteritidis was 424 out of 492 strains (86.2%). PMID- 8708412 TI - [A case of Pasteurella multocida subsp. multocida septicemia due to cat bites in liver cirrhosis patient]. AB - A 60-year-old male who had been suffering from liver cirrhosis was admitted to our hospital with high grade fever accompanied by right chest pain. Chest X-rays revealed a moderate amount of pleural fluid suggesting pleuritis. Multocida was isolated from the blood culture as well as the pleural fluid. Antibiotic therapy was initiated according to the drug susceptibility of the isolates. Ten days treatment was effective on the cessation of both septicemia and the clinical symptoms. Since the patient had been bitten several times by his own pet cats, their mouth swabs were taken for pathogenic investigations. Serotypes of the cats' isolates coincided with that of the patient's which consequently indicated the route of infection. P. multocida is a Gram negative coccobacillary organism that resides as normal flora in the oral cavity of animals, including dogs and cats. It has been originally known to be a causative agent for hemorrhagic septicemia in domestic animals. However, recently reports of P. multocida infections in man has been increasing due to the enlargement of pet populations. Although outbreaks of septicemia is rare, it occurs most often in immunologically compromised hosts, including patients with liver cirrhosis as in this case. Therefore, it is important to initiate an urgent antibiotic therapy in such cases. Overall, it is of utmost importance to instruct immunosuppressed patients to avoid excessive exposure to animals including pets. PMID- 8708413 TI - [A case report of invasive pulmonary aspergillosis which could not be saved in spite of making an early diagnosis]. AB - A seventy-one year old male with pemphigus vulgaris and treated with steroid therapy for long periods of 7 months was suffered from invasive pulmonary aspergillosis. He complained of a productive cough and the chest X-p indicated a mild infiltration shadow in his both lungfields. He was treated by intravenous antibiotics and no clinical improvement was observed. Two days after the onset of the clinical respiratory symptoms, he was transferred to the division of respiratory diseases and the diagnosis of aspergillosis was confirmed by serological and histopathological studies on the same day. Intravenous amphotericin B and oral itraconazole administrations were started immediately after the diagnosis was made. However, the progression of the disease was so rapid and his immune condition was too weak to respond to the treatment. The overall clinical course of the case was extremely short only 5 days. PMID- 8708414 TI - [A case of toxoplasmosis with dermatomyositis]. AB - We report a case of dermatomyositis (DM) in a 15-year-old female with toxoplasmosis after ingestion of raw bovine liver. Facial erythema and cervical lymphadenopathy preceded myalgia and muscle weakness of the extremities. The diagnostic criteria of DM was fulfilled because of symmetrical and proximal dominant muscle weakness, elevation of myogenic enzyme (CPK, GOT, LDH, myoglobin, aldorase), myogenic pattern of electromyogram, skeletal muscle biopsy showing interstitial myositis with mild destruction of muscle fiber, and facial erythema. Immunological findings showed IgG anti-toxoplasma antibody to be 1340 IU/ml and IgM to be 7.0 (Cut off index 0.7), suggesting acute toxoplasmosis. Treatment with prednisolone for DM and acetylspiramycin for toxoplasmosis was successful. Toxoplasmosis should be considered as a possibility in patients with myositis. PMID- 8708415 TI - [Src family of protein-tyrosine kinases and their regulator tyrosine kinase, Csk]. PMID- 8708416 TI - [Affinity electrophoresis--its application for studies on heterogeneity and functions of biomolecules]. PMID- 8708417 TI - [The analysis of maternal inheritance of mitochondrial DNA in mammal]. PMID- 8708418 TI - [Recent developments in studies on transcription factors--role of TFIID in transcriptional activation]. PMID- 8708419 TI - [8-Oxoguanine repair system and its role in mitochondria]. PMID- 8708420 TI - [Biochemical aspects of the telomere]. PMID- 8708421 TI - [Gas6: a ligand for receptor tyrosine kinase containing gamma-carboxyglutamic acids]. PMID- 8708422 TI - [Simple method for detection of sense and antisense transcripts]. PMID- 8708423 TI - Vascular surgery in the European Union: current state, developments and prospects for the future. PMID- 8708424 TI - Vascular risk factors for atherosclerotic plaque progression in carotid artery. AB - Two-dimensional ultrasound is a widely used technique for the clinical assessment of carotid atherosclerosis. We measured volumes of atherosclerotic plaques in carotid arteries in 70 patients with a newly developed and tested three dimensional ultrasound method over a period of twelve months to determine the relation between progression or regression of plaque volume and vascular risk factors. In multiple regression analysis of age, size, smoking, total cholesterol, HDL-cholesterol, HDL/total cholesterol ratio, LDL-cholesterol, triglycerides, HbA1, diastolic and systolic blood pressure, the strongest predictor of plaque progression (p < 0.01) was diastolic blood pressure, diabetes came next (p < 0.05). The optimal adjustment of all risk factors showed a significant influence on plaque progression (r: 0.76). PMID- 8708425 TI - Relation of haemostatic, fibrinolytic, and rheological variables to the angiographic extent of peripheral arterial occlusive disease. AB - We investigated the relationships between the angiographic severity of peripheral arterial occlusive disease (PAOD) and haemostasis, fibrinolytic, and rheological variables in 219 patients with symptomatic peripheral arterial occlusive disease (PAOD). White cell count, fibrinogen, cross-linked fibrin degradation products (FDP), von Willebrand factor, and plasminogen activator inhibitor levels were all elevated in comparison with age-matched population controls (all p < 0.0001, Mann Whitney U test), while fibrinogen (Spearman r = 0.30), von Willebrand factor (r = 0.40), and log (FDP) (r = 0.56), (all p < 0.0001) showed a strong correlation with the angiographic extent of PAOD. Multivariate analysis indicated that log (FDP) was a strong independent predictor of the angiographic severity of PAOD (p < 0.0001), in addition to increasing age (p < 0.0001), presence of tissue sepsis (p < 0.02), prior vascular surgery (p = 0.007), and other vascular pathology (p = 0.007). These results confirm that increase in fibrinogen, von Willebrand factor, plasminogen activator inhibitor and fibrin turnover, are strongly associated with the presence of symptomatic peripheral arterial disease, and suggest that there may be a causal link between fibrin turnover, as determined by FDP levels, and the extent of peripheral arterial occlusive disease. PMID- 8708426 TI - Critical limb ischaemia: a case against Consensus II. AB - Critical limb ischaemia (CLI) is a serious clinical condition that often immediately precedes limb loss. The Consensus Documents of 1989 and 1991 attempted to define CLI and give direction to its investigation and management. Whilst the need for such a consensus was clear and should be supported we believe the definition of CLI as documented in the Consensus Documents I and II recommendation number I is wrong. We present evidence from 140 patients with severe limb ischaemia taken from the PARTNER Group studies to support our request for an amendment to the ankle pressure recommendation from < or = 50 mmHg to >50 mmHg and big toe pressure from < or = 30 mmHg to >30 mmHg for the purpose of conducting clinical trials and to include Doppler index and tcPO2 as additional parameters. We also believe that the current document may be actually excluding the only group of patients likely to benefit from drug treatment or other interventions and that the above amendment should be prioritized. PMID- 8708427 TI - Lipoprotein(a) and general risk factors in patients with angiographically assessed peripheral arterial disease. AB - High lipoprotein(a) [Lp(a)] has been observed in patients with ischemic heart disease and cerebrovascular disease. Lp(a) is actually thought to be an independent risk factor for coronary disease. We therefore carried out a case control study, evaluating plasma Lp(a) in 61 patients with angiographically documented peripheral arterial disease (PAD) and in 61 age- and sex-matched patients with no cardiovascular disease. General risk factors for vascular disease were also taken into account. Lp(a) was significantly higher in patients than controls (257.0 +/- 34.8 vs 146.5 +/- 23.5 mg/l p < 0.05), as were cigarette smoking, diabetes, cholesterolemia, fibrinogenemia and the waist-to-hip circumference ratio. Stepwise logistic regression analysis showed that, in addition to cigarette smoking, diabetes, cholesterol and fibrinogen, Lp(a) is a significant independent risk indicator for PAD. This result suggests that high plasma Lp(a) is associated with enhanced risk of PAD and must therefore be evaluated alongside traditional risk factors. PMID- 8708428 TI - "Kissing-balloon" technique for abdominal aorta angioplasty. Initial results and long term outcome. AB - We evaluated results after angioplasty of the distal abdominal aorta using "kissing-balloon" technique. MATERIALS AND METHODS. From 1981 through 1993, 14 patients (12 women and 2 men, average age 52.4 years), underwent balloon angioplasty of the distal segment of the abdominal aorta. In 8 patients stenosis involved one or both iliac arteries as well. In all cases double balloon technique was used. Follow-up was based on clinical, angiographical and duplex scanning examinations. RESULTS. The initial success rate was 85% (12/14). In one case there was residual stenosis of 20% (redilated successfully 1 month later), and in one case there was acute thrombosis of the right iliac artery (successfully relieved surgically). Long-term follow-up (6-143 months) showed good patency of the dilated segment in all patients (including patients that initially failed). At follow-up, all patients were asymptomatic. CONCLUSION. Dilatation of the distal abdominal aorta using "kissing-balloon" technique is a safe and efficient modality of treatment, with acceptable initial and excellent long-term results. PMID- 8708429 TI - Clinical aspects and treatment of inflammatory abdominal aortic aneurysms. AB - An increasing number of inflammatory abdominal aortic aneurysms are reported today, although it is not clear whether these are nosologically independent lesions or a particular evolution of atherosclerotic aneurysms with enhanced phlogistic processes and fibrous reaction. Several pathogenetic theories have been proposed (microfissuration, autoimmunity, lymphatic stasis); clinical symptoms are characterised by the frequent involvement of contiguous retroperineal structures (in particular the urinary excretory tract and duodenum) which may be dislocated or compressed by the neoformation. CT or NMR appear to be the instrumental tests which give the most reliable diagnosis regarding the suspected inflammatory nature of the aneurysm. A correct pre-operative diagnosis is particularly important given that the morphological peculiarities (fibrosis and tenacious synechiae) add considerable pitfalls and difficulties to the operation. In this respect, the use of special technical devices, such as minimum dissection, permit a marked reduction of perioperative complications. PMID- 8708430 TI - Inhibition of intimal hyperplasia and occlusion in Dacron graft with heparin and low molecular weight heparin. AB - OBJECTIVE: Intimal hyperplasia (IH) is a significant cause of late prosthetic graft occlusion. The influence of heparin and low molecular weight heparin (LMWH) on IH was studied in a sheep model. EXPERIMENTAL DESIGN: A gelatin sealed Dacron (Gelsoft) interposition vascular graft was placed end to end in sheep carotid artery. Standard heparin (SH) was administered in a dose of 7,500 units twice daily subcutaneously (n = 6). LMWH (enoxaparin) was administered in two dosages, a "low dose" of 2 mg/kg (n = 6) and a "high dose" of 5 mg/kg (n = 6) subcutaneously daily. A control group received a graft but no heparin (n = 6). At one month the animals were sacrificed and the grafts sectioned. An intimal measurement was obtained under light microscopy using an eyepiece graticule from a longitudinally sectioned graft. RESULTS: Controls occluded five grafts of six and recorded a mean intimal thickness of 2,510 micrometres, SH sheep occluded two with a mean thickness of 83 (p < 0.05), low dose enoxaparin sheep had no occlusions with a mean thickness of 73 (p < 0.001) and high dose enoxaparin sheep occluded one with a mean thickness of 225 (p < 0.01). The reduced occlusion rates were significant for the LMWH groups (p = 0.007 for low dose, p = 0.04 for high dose) but not for SH. There were no significant differences between treatment groups with respect to IH. CONCLUSIONS: In this sheep animal study, each of SH and two LMWH dose regimens resulted in a reduction in Dacron vascular graft occlusion rate and the development of IH, with the greatest effect seen in the low dose LMWH group. PMID- 8708431 TI - The value of quantitative bacteriological investigations in the monitoring of treatment of ischaemic ulcerations of lower legs. AB - The quantitative and qualitative bacteriological investigations of 63 patients were done on ischaemic ulcerations before reconstructive vascular surgery and at 7 day intervals after the operation. Among the isolated bacteria the most common were Gram positive (62.9%), especially Staphylococcus aureus. Amputations due to non-healing ulcers were performed on 8 patients, who had ankle brachial index (ABI) lower than 0.47. In 55 patients with ABI higher than 0.47 (with the exception of one case) free skin grafts were applied to reduce the time of the ulcers healing. Primary healing of ulcers covered with free-skin grafts was achieved in 44 out of 55 patients (80%). In 11 patients, were free-skin grafts had failed, ulcerations were healed following the repetition of the free-skin grafts. The healing results of skin grafts statistically were significantly better in the group where the number of bacteria in 1 cm2 of ulceration was lower than 50.000. The severity of infections in ulcers makes the healing process of skin grafts impossible. Quantitative bacteriology additionally helps in objective evaluation of granulating tissue and facilitates choice of proper skin grafting time. This study has shown the usefulness of quantitative bacteriology for the determination of the severity of infections in ulcers. PMID- 8708432 TI - Oxidative stress is present in atherosclerotic peripheral arterial disease and further increased by diabetes mellitus. AB - OBJECTIVE: Atherosclerosis, which is associated with increased oxidative stress, may be widespread and more advanced in both complicated and uncomplicated diabetics than in healthy, age-matched non diabetic controls. The development of microangiopathy and the early development of microangiopathy and the early development of macroangiopathy which occurs in diabetes have unclear aetiology. We have previously shown an increase in the products of free radical activity in conventional atherosclerosis and this study has considered whether there us a further increase in this oxidative stress in diabetics. EXPERIMENTAL DESIGN: We measured oxidation by free radicals as lipid peroxides (malondialdehyde) and plasma thiols in 19 insulin dependent diabetic patients with retinopathy or microproteinuria but no clinical evidence of large vessel disease, 19 non diabetic patients with obstructive peripheral arterial disease confirmed by angiography and 19 asymptomatic matched healthy control subjects. PATIENTS: The patients were recruited from either the local diabetic or vascular surgery clinics. RESULTS: There were significant increases in oxidative stress in diabetic patients compared to control subjects with increased malondialdehyde 9.2 mumol/l (7.1-11.2) versus 7.65 mumol/l (5.4-9.6, p < 0.01 Mann Whitney test) and reduced plasma thiols 458 mumol/l (294-595) versus 506 mumol/l (433-589, p < 0.05). Those non-diabetic patients with obstructive peripheral arterial disease also had significantly increased malondialdehyde levels at 8.4 mumol/l (6.5-10.1, p < 0.05) and reduced plasma thiols 435 mumol/l (361-462, p < 0.05) compared to controls. The difference between patients with conventional atherosclerotic peripheral arterial disease and those with additional diabetes was significant for malondialdehyde (p < 0.05). CONCLUSIONS: These data confirm the oxidative stress that exists in atherosclerotic peripheral arterial disease and suggest that diabetic patients with microangiopathy have an increase in this oxidative stress which may contribute to their early development of atherosclerosis. PMID- 8708434 TI - Tilting enhances visualization of lower extremity veins with duplex colour flow imaging. AB - The present paper presents the results of a comparative analysis of the number veins and of the ease of visualisation of below the knee as displayed by colour Duplex scanning in both legs of 10 normal individuals in the horizontal and 40 degrees foot down tilted position. Foot-down tilting improves the ease of visualisation of lower leg veins by 38% and the number of visible branches by 28%. Therefore, this technique should preferably be employed in the diagnosis of deep vein thrombosis in order to improve diagnostic accuracy. PMID- 8708433 TI - Hydrotherapy of patients with intermittent claudication: a novel approach to improve systolic ankle pressure and reduce symptoms. AB - OBJECTIVE: To study the effects of alternating cold and hot water therapy on walking ability and systolic blood pressure in claudicants. EXPERIMENTAL DESIGN: A prospective case study with repeated measurements before and 1, 4 and 12 months after treatment. The systolic blood pressure levels were measured with an occlusion cuff for brachial and ankle and with a strain gauge for the first toe. STUDY POPULATION: Twenty consecutively included patients, 11 women and 9 men; mean age 73.9 yrs, with intermittent claudication according to clinical examination and ankle-arm systolic blood pressure (AAI) below 0.90. INTERVENTION: Alternate hot and cold hydrotherapy of the legs were given at ten 25-minute treatments during a three-week period. The outcome measures were maximal walking ability (MW), walking ability before pain (PW) and systolic blood pressures of toe, ankle, arm and AAI. RESULTS: Fourteen patients (70%) reported reduced pain after treatment and their PW increased from 134 +/- 29 m to 415 +/- 119 m 12 months later (p < 0.05) and the MW in the total group increased form 348 +/- 75 m to 523 +/- 103 m. Systolic blood pressure increased in right ankle and toe one month after treatment in the total group. Among those who reported improved walking ability one year after treatment, systolic blood pressure in both right and left ankles and toes increased; e.g. right toe increasing from 72 +/- 7 to 86 +/- 2 (p < 0.001). Improvements of systolic blood pressure in left and right leg and changes of walking ability were correlated, in the order of 0.60 to 0.81, p < 0.05. CONCLUSIONS: Showering the legs of claudicants improved walking ability and blood pressures which sustained up to 1-year later. This therapy might be an additional alternative to conservative treatment of intermittent claudication. PMID- 8708435 TI - Outpatient surgery of varices of the lower limbs: experience of 2,568 cases at four universities. AB - Outpatient surgery of varices of the lower limbs is currently considered a viable alternative to traditional surgery with hospitalization. This paper reports the experience of 4 university groups (Padua, Modena, Verona, Milan), where outpatient treatment has been used since 1987. From June 1987 to June 1992, 2,568 lower limb varices were treated in this way. Different techniques of anaesthesia were used (local infiltration, combined local and general, general, subarachnoid). In all cases, crossectomy was combined with short or long saphenous stripping. There were no intra- or perioperative deaths, and only limited morbidity. Postoperative hospitalization was required in only 2 cases: for hemorrhaging of the inguinal wound in one case, and headache 2 days after spinal anaesthesia in the other. In 2 separate samples of 100 patients, 88 and 89 indicated satisfaction with the surgical treatment. In conclusion, outpatient surgery of varices can be based on the same techniques as in-patients treatment. The risks of surgery and anaesthesia in specialised centres are very limited, with scope for a variety of anaesthetic techniques according to facilities available. Patients satisfaction is high. PMID- 8708436 TI - Clinical-anamnestic and instrumental data in outpatients suffering from venous disease. AB - The authors studied 200 consecutive outpatients with symptoms due to venous disease of the lower limbs. All the patients underwent anamnestic, clinical and instrumental evaluation (c.w. Doppler velocimetry and strain-gauge plethysmography). The results of this study showed that some anamnestic and clinical data are significantly related to venous disease detected by instrumental techniques. The superficial and deep vein hypertension were, instead, correlated to oral contraceptive use. PMID- 8708437 TI - Transaxillary first rib resection for thoracic outlet syndrome: follow-up of 9 cases. AB - Nine cases of Thoracic Outlet Syndrome (TOS) were treated by transaxillary 1st rib resection over a three year period. Mean follow-up was 26 +/- 4 months. Patients were recalled every 3 months post-operatively for surveillance. Parameters for evaluation included physical examination, return of patient to (full-range or partial range) pre-illness activity, nerve conduction velocities, somato-sensory evoked potentials, Duplex imaging and venography or arteriography if the latter two were indicated. Neurogenic involvement with or without vascular compression was present in 7 cases (78%) and satisfactory results of surgery were recorded in 6 patients (67%). The clinical settings and detailed results are reviewed and variable factors that might alter the outcome are analyzed in this series. Neither duration of symptoms, sex differences nor associated bony anomalies had any effect on operative results. The single factor that seemingly had a negative impact on the outcome after surgery was the patient's need to return to strenuous and repetitive arm and/or hand movements. PMID- 8708439 TI - Raeder's syndrome. Report of two cases. AB - Raeder's syndrome constitutes facial pain and ipsilateral stenosis of the palpebral fissure, miosis and enophthalmos. It is divided into two groups. Group I includes cases with parasellar cranial nerve involvement and group II without parasellar cranial nerve involvement. It is often difficult to distinguish group II of Raeder's syndrome from Horner's syndrome. The latter is painless and is accompanied by anidrosis of the ipsilateral half of the face. In fact Raeder's syndrome may be caused by any lesion affecting the post-ganglionic oculosympathetic fibers distal to the bifurcation of the common carotid artery. Serious lesions of the internal carotid artery (ICA) may be responsible for this syndrome. This paper deals with two cases of group II of Raeder's syndrome. In the first case, a 60-year-old man, the angiogram revealed an aneurysm of the extracranial part of the ICA just below its entrance into the calvarium. In the second case, a 42-year-old man, a dissective aneurysm of the ICA was found. Surgery was not attempted in either of our patients for different reasons. Their symptoms ameliorated quickly with medical treatment. PMID- 8708438 TI - Reevaluation of duplex criteria for diagnosis of high-grade carotid artery stenosis: when do we need angiography and when can we operate without angiography? PMID- 8708440 TI - [Concept and definition of asthma]. PMID- 8708441 TI - [Epidemiology of asthma (occurrence and death)]. PMID- 8708442 TI - [Mechanism of airway inflammation in patients with asthma]. PMID- 8708443 TI - [Inflammatory mediators in asthma]. PMID- 8708444 TI - [Role of cytokines in pathophysiology of asthma]. PMID- 8708445 TI - [Airway inflammation in asthma]. PMID- 8708446 TI - [Diagnosis of bronchial asthma]. PMID- 8708447 TI - [Guidelines for the treatment of asthma]. PMID- 8708448 TI - [Steroid inhalation therapy of asthma]. PMID- 8708449 TI - [Therapy of asthma using bronchodilator agents--special reference to adrenergic beta 2-receptor agonists]. PMID- 8708450 TI - [Anti-allergic agent therapy of asthma]. PMID- 8708451 TI - [Management of asthma based on their peak expiratory flow rate monitoring]. PMID- 8708452 TI - [Aspirin-induced asthma]. PMID- 8708453 TI - [Occupational asthma]. PMID- 8708454 TI - [Cough variant asthma]. PMID- 8708455 TI - [Adult onset of asthma and their therapy (discussion)]. PMID- 8708456 TI - [Autopsy case of angiotropic lymphoma complicated with interstitial pneumonia during VEPA therapy and monocytosis in the early stage]. PMID- 8708457 TI - [Case of Fisher syndrome with pemphigus treated by immuno-adsorption plasmapheresis]. PMID- 8708458 TI - [A case of various cardiac disorders due to interferon-alpha]. PMID- 8708459 TI - [A case of intractable myasthenia gravis treated with cyclosporine and high-dose intravenous immunoglobulin]. PMID- 8708460 TI - [A case of eosinophilic myocarditis presenting with edematous thickening of endocardium]. PMID- 8708461 TI - [Insulin-resistance syndrome]. PMID- 8708462 TI - [Penicillin-resistant Streptococcus pneumoniae]. PMID- 8708463 TI - [Study on diabetes mellitus--recent progress]. PMID- 8708464 TI - [Progress on clinical study and prediction of IDDM]. PMID- 8708465 TI - [Progress on clinical and etiological study of NIDDM]. PMID- 8708466 TI - [Therapy of diabetes mellitus associated with hypertension]. PMID- 8708467 TI - [Therapy of diabetes mellitus associated with hyperlipidemia]. PMID- 8708468 TI - [Therapy of diabetes mellitus associated with liver disease]. PMID- 8708469 TI - [Diabetes mellitus with endocrine diseases--the physiopathology , diagnosis and therapy]. PMID- 8708470 TI - [Diabetes mellitus during pregnancy]. PMID- 8708471 TI - [Progress on diagnosis and therapy of diabetic retinopathy]. PMID- 8708472 TI - [Progress on diagnosis and therapy of diabetic nephropathy]. PMID- 8708473 TI - [Progress on diagnosis and therapy of diabetic neuropathies]. PMID- 8708474 TI - [Progress on diagnosis and therapy of diabetic gangrene]. PMID- 8708475 TI - [Progress on diagnosis and therapy of diabetic angiopathies]. PMID- 8708476 TI - [Pathology of diabetes mellitus]. PMID- 8708477 TI - [Mitochondria gene mutation and diabetes]. PMID- 8708478 TI - [Obesity and diabetes mellitus]. PMID- 8708479 TI - [Progress on clinical study of diabetes mellitus (discussion)]. PMID- 8708480 TI - [A case of Werner's syndrome associated with heart failure due aortic valve stenosis]. PMID- 8708481 TI - [A case of Evans's syndrome with antiphospholipid antibody syndrome]. PMID- 8708482 TI - [A case of systemic lupus erythematosus with pleural effusion, ascites and pericardial effusion]. PMID- 8708483 TI - [A case of Mounier-Kuhn syndrome]. PMID- 8708484 TI - [A case of acute progressive glomerulonephritis with positive P-ANCA and C-ANCA]. PMID- 8708485 TI - [Epstein-Barr virus and hematologic diseases]. PMID- 8708486 TI - [Neuropathy and anti-ganglioside antibodies]. PMID- 8708487 TI - Analyses of data quality in registries concerning diabetes mellitus--a comparison between a population based hospital discharge and an insulin prescription registry. AB - To evaluate the data quality in the Danish National Registry of Patients (DNRP) and the Prescription Registry in the country of Northern Jutland (487,000 inhabitants) concerning insulin dependent diabetes mellitus (IDDM) and insulin treated diabetes mellitus, a comparison between data in the two registries was made. From the Regional Hospital Registry in the County of Northern Jutland, containing discharge diagnoses from all admissions to hospitals in the county, we identified all patients with the IDDM diagnosis between 1987 and 1993. From the Regional Prescription Registry all insulin prescriptions taken up at pharmacies in the county in 1993 were identified. All persons were identified by their individual identification number (CPR-number), and a record linkage between the two data sources was made. The predictive value of an IDDM-registration in the DNRP was 96% and the corresponding completeness 91%. In the Prescription Registry the completeness was 96%. Both registries seem to be valuable study bases for epidemiological research in diabetes mellitus. PMID- 8708488 TI - Order determination in autoregressive modeling of diastolic heart sounds. AB - Previous studies demonstrated that spectral analysis of diastolic heart sounds may provide valuable information for the detection of coronary artery disease. Although parametric modeling methods were successfully used to achieve this goal, and showed considerable performance, the accuracy and precision of the analysis is strongly dependent on the model order selected. In order to investigate the effects of model order selection on the analysis, diastolic heart sound recorded from both normal and diseased patients were analyzed using the AR modeling, which is computationally the most efficient parametric spectral analysis method. The model order were determined by using four different model order selection criteria. The results showed that the four criteria yielded different order for the same data set. On the other hand, different criteria showed different performance in different measurement conditions. Effect of arbitrary order selection was also discussed. As a result, an optimal AR model order that may be used for every case was determined. PMID- 8708489 TI - "Compu-Void II": the computerized voiding diary. AB - OBJECTIVE: We have previously described an electronic voiding diary, "Compu-Void" (Copyright, 1990) developed to automate recording of bladder symptoms (Rabin et al., 1993). Our objectives in this, the second phase of this study, were to examine a group of subject and control patients' preference and compliance with regard to the "Compu-Void" (CV) compared to the standard written voiding diary (WD), to compare the two methods with respect to the amount and type of information obtained and to determine whether or not the order of use of each recording method influenced results in the subject group. METHODS: Thirty-six women between the ages of 20 and 84 with bladder symptomatology were compared to a group 36 age-matched women. RESULTS: In 100% of subjects and 95% of control patients, CV entries exceeded the number made with the WD in voiding events and in subjects, in incontinent episodes recorded (P < 0.0005 and P < 0.005, respectively). Over 98% of subjects and over 80% of control patients preferred CV over the WD (p < 0.0005). The order of use of each recording method in subjects made no significant difference with regard to the volume of information obtained (p < 0.407), number of urinary leakage events recorded (p < 0.494), and fluid intake patterns (p < 0.410). Patient impressions of, and compliance with each method were not affected by order of use. The only difference regarding order of use was that most subjects who used the CV first also found the WD to be tedious (61% vs 14%). CONCLUSIONS: Our results suggest increased volume of data and of patient compliance in reporting bladder symptoms and events using CV, and that order of use is not an important factor in determining patient impressions of the two methods. The majority of subject and control patients preferred CV over traditional methods. An updated version of the software and hardware is also included. PMID- 8708490 TI - An ECG storage and retrieval system embedded in client server HIS utilizing object-oriented DB. AB - In the University of Tokyo Hospital, the improved client server HIS has been applied to clinical practice and physicians can order prescription, laboratory examination, ECG examination and radiographic examination, etc. directly by themselves and read results of these examinations, except medical signal waves, schema and image, on UNIX workstations. Recently, we designed and developed an ECG storage and retrieval system embedded in the client server HIS utilizing object-oriented database to take the first step in dealing with digitized signal, schema and image data and show waves, graphics, and images directly to physicians by the client server HIS. The system was developed based on object-oriented analysis and design, and implemented with object-oriented database management system (OODMS) and C++ programming language. In this paper, we describe the ECG data model, functions of the storage and retrieval system, features of user interface and the result of its implementation in the HIS. PMID- 8708491 TI - A prototype computer decision support system for the management of asthma. AB - Asthma is a chronic disease estimated to affect 6-7% of the total UK population. In addition, a number of studies have shown that asthma has become commoner since the 1970s, especially in children. The diagnosis of asthma can be difficult and its management requires the involvement of patients in a long-term treatment plan, something which general practitioners may be unable to achieve easily in the average 10-min consultation. As a consequence, asthma is underdiagnosed and undertreated. Deaths from the disease are often avoidable with timely and sufficient use of the available medication. In order to support this, the British Thoracic Society (BTS) has published guidelines for asthma management based upon a stepwise approach, in which a patient is categorized as being on one of five steps according to the severity of his or her asthma. The guidelines give "rules of thumb" for deciding when the patient should move up or down the steps. The most recent version of the guidelines also included special rules for children. Within a recent European Community project on Advanced Informatics in Medicine (AIM), we developed a prototype decision support system for asthma management targeted at the primary care setting and based on the British Thoracic Society guidelines. This paper reports this development, and describes the further work needed on the prototype. Plans for evaluation of the knowledge bases and for future full application production are also described. PMID- 8708492 TI - Identification of patients treated with strong analgesics: an assessment of two Danish information systems with respect to epidemiological research. AB - Pain is a significant health problem, and there is considerable need for clinical and epidemiological research in this topic. A prerequisite for doing research on patients treated with strong analgesics is that it is possible to identify the patients. We assessed two Danish population-based information systems, in which patients treated with strong analgesics are registered by using the patients' personal registration numbers as identifier. The two systems, which we compared, were (1) a surveillance system administered by the National Board of Health, and (2) the drug prescription register in the Danish National Health Service. During August 1994, 3787 patients were registered in the surveillance system and 3812 in the National Health Service in North Jutland County. Ninety-five persons were registered only in the surveillance system, and 120 only in the National Health Service register. A capture-recapture analysis showed a coverage of 96.9% for the surveillance system and 97.5% for the National Health Service. We thus conclude that the two systems form a valuable study base of patients treated with strong analgesics in epidemiological research. PMID- 8708493 TI - Helicobacter pylori infection in Thai children with recurrent abdominal pain. AB - Sixty-five children with recurrent abdominal pain underwent gastrointestional endoscopy which showed Hp gastritis without duodenal ulcer in 16.9 per cent of cases. The prevalences of infection in recurrent abdominal pain and asymptomatic children were not different. Either urease test or histological method was appropriate for diagnosis of this infection. Triple therapy including bismuth subcitrate, amoxicillin and metronidazole improved abdominal pain symptom in 72.7 per cent without any side effect. PMID- 8708494 TI - Detection of HIV-1 proviral DNA by polymerase chain reaction: a preliminary study in Bangkok. AB - Nested polymerase chain reaction (nested PCR) was used to separately amplify part of gag, pol, and env genes of human immunodeficiency virus type 1 (HIV-1) to evaluate that primer specific to either gag (SK380/390&SK38/39), pol (JA17/18&JA19/20), or env (JA9/10&JA11/12) genes is suitable for HIV-1 PCR based diagnosis in Thailand. The positive PCR results in 70 HIV-1 infected adults are 100, 97, 89 per cent and in 75 HIV-1 infected infants are 100, 94, 74 per cent by gag, pol, env primer, respectively. The specificity of all three primer sets is 100 per cent. The unamplified samples by pol and env primers were identified as HIV-1 subtype E by PELISA method. False negative in HIV-1 PCR based diagnosis caused by high genetic variation of HIV-1 can be overcome by using several primer sets as shown in this study. PMID- 8708495 TI - Effect of paper spacer (spader) in conjunction with procaterol metered dose inhaler in the treatment of acute asthma. AB - Thirty acute asthmatic patients (mean age 35.6 +/- 13.5 years, mean FEV1 0.97 +/- 0.95 L) in the emergency room were evaluated for airway response to procaterol administered by metered-dose inhaler (MDI) compared to MDI with auxiliary device (a paper spacer) in a randomised cross over study. Bronchodilator response to procaterol delivered by MDI, with paper spacer was significantly greater than that by MDI without spacer (mean maximal increment of FEV1, 0.67 +/- 0.46 vs 0.59 +/- 0.46 L, p = 0.016, mean maximal increment of PEFR, 112 +/- 62.3 vs 86.7 +/- 61.3 L/min, p = 0.003) but there was no difference in dyspnoea score, wheezing score and heart rate. This collapsible and portable auxiliary device (paper spacer) can increase therapeutic efficacy of bronchodilator response in asthmatic patients. PMID- 8708496 TI - Comparison of oral rehydration solutions made with rice syrup solids or glucose in the treatment of acute diarrhea in infants. AB - An oral electrolyte solution made with rice syrup solids was found to reduce fecal output more effectively than a comparable glucose-based solution in the early hours of treatment of infantile diarrhea. Administration of the rice syrup solids solution was also more effective in promoting absorption of fluid, sodium, and potassium. In some infants, the rapid, effective rehydration offered by this solution may prevent the need for hospitalization. PMID- 8708497 TI - Homograft myringoplasty-controlled button technique: a preliminary report. AB - The successful outcome of homograft myringoplasty in small and medium perforation of the tympanic membrane was only 44.12 per cent. A better result was obtained in the small perforations (59.10%) compared with the medium ones (16.77%). We conclude that the procedure should be appropriate for repairing the incomplete healing of the tympanic membrane following standard tympanoplasty and removal of the tympanostomy tube. Furthermore this procedure is simple, safe, cost-effective and can be done in the out-patient department. PMID- 8708499 TI - Cardiovascular and respiratory changes in laparoscopic cholecystectomy. AB - Although laparoscopic cholecystectomy has several advantages over the traditional open cholecystectomy, it may however cause some adverse cardiovascular and respiratory effects associated with intraperitoneal insufflation of CO2. It is, therefore, recommended that appropriate hemodynamic and respiratory monitorings should be used routinely in order to prevent deleterious complications, especially in high risk patients. PMID- 8708498 TI - Methacholine inhalation challenge in patients with chronic cough induced by angiotensin-converting enzyme inhibitors. AB - Angiotensin-converting enzyme inhibitors (ACEI) cause cough in some patients, but the mechanism of this side effect is not clear. Five patients (group I) who had developed chronic cough induced by ACEI were evaluated to determine the bronchial hyperreactivity (BHR) by challenge with methacholine inhalation using a reservoir method at Respiratory Unit, Chulalongkorn Hospital University. Five patients (group II) who did not experience coughing associated with ACEI were also challenged as controls. The results revealed that two patients (40%) in group I demonstrated BHR with the mean PC20 at 15 mg/ml of methacholine solution. On the other hand, none of the patients in group II disclosed BHR. We concluded that coughing during ACEI therapy may be due to an increased inflammatory state in the airway in some susceptible subjects. PMID- 8708500 TI - Surgical anatomy of lateral nasal wall in northeast Thai cadavers. AB - A morphometric osteological study of the lateral nasal wall of Thai cadavers proved essential for the endoscopic sinus surgery on Thai patients in order to increase effectiveness and decrease the number of intraoperative complications. The middle turbinate should be preserved during the operation because it is a useful landmark for finding the natural maxillary sinus ostium and for avoiding nasolacrimal duct injury. PMID- 8708502 TI - Rett syndrome: a case report. AB - Rett syndrome (RS) is a progressive neurodegenerative disorder characterized by almost exclusive occurrence in females, autistic behavior, dementia, gait ataxia, loss of purposeful use of the hands with stereotypic hand movement. Initially RS was considered to be very rare; however, recent reports suggest that the prevalence is considerably higher and occurrence is world-wide. There are as yet no pathognomonic laboratory markers and its pathogenesis and etiology are unknown. We report a 4-year-old girl who fulfilled the clinical criteria of classic Rett syndrome. PMID- 8708501 TI - Chemoprevention of head and neck cancer. AB - Tobacco-related upper aerodigestive tract cancer is a worldwide health problem. Clinical advances in surgery, radiation, and chemotherapy have shown only marginal improvement on the survival rate of these malignancies in the past two decades. The concept of multistage carcinogenesis and field cancerization, the high incidence of SPTs, and the promising results of chemopreventive agents especially 13 cRA to effectively reverse oral premalignancy and prevent SPTs indicate a new avenue to improve the outcome of these newplastic diseases. The study of second primary tumor prevention is too early to show any survival benefit in the treatment arm and toxicity of high dose 13 cRA is significant. The current prospective large scale phase III SPT prevention trial of low-dose 3 cRA may establish a new standard approach for the control of head and neck neoplasms. Identification of potentially useful panels of biomarkers as intermediate endpoints will undoubtedly reduce time and cost currently required to conduct chemoprevention trials. PMID- 8708503 TI - Primary ovarian carcinoid: a mixture of insular, trabecular and mucinous components. AB - A case of primary ovarian carcinoid in a 53-year-old woman is described. The tumor which replaced the left ovary showed a mixture of insular, trabecular and mucinous cell types in the same mass. The nature of the neoplastic cells has been verified by light and electron microscopic studies. The latter investigation demonstrated bundles of tonofilament and apical villi of the trabecular cells in addition to the morphologic difference of neurosecretory granules among these various cells. PMID- 8708504 TI - An open label trial of pergolide in Thai patients with Parkinson's disease. AB - Fifteen Thai patients with Parkinson's disease (7 females, 8 males) were enrolled in an open label trial of pergolide (a new dopamine agonist) to evaluate its safety and efficacy. Inpatients and outpatients from Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand from 1992 to 1994 were included in the study with a total duration of 18 weeks. Both de novo patients and patients who were being treated with levodopa without dopamine agonist and were obtaining a less than optimal response at both visit 1 and visit 2 were all enrolled in this study. At entry into the study, 3 patients had Hoehn and Yahr stage I, 7 patients at stage II, 3 patients at stage III, and 2 patients at stage IV. Pergolide dosage was gradually built up until an optimal dosage was achieved. The average dose of pergolide during the study was 0.94 mg/day (range 0.075 to 8 mg/day). All patients completed the study and no patients dropped out. Two patients (13.33 per cent) experienced nausea (on 0.4 mg/day and 0.075 mg/day), two patients (13.33 per cent) experienced sleepiness (0.50 mg/day and 0.075 mg/day) and one patient (6.67 per cent) unsteadiness on walking (0.50 mg/day). There was one patient who required pergolide up to 8 mg/day which is higher than the recommended dosage (5 mg/day) but this patient experienced no adverse effects and his disabled dyskinesic was abolished. Our study demonstrated the good toleration and efficacy of pergolide treatment for Thai patients with Parkinson's disease. This new dopamine agonist stimulates both D1 and D2 receptors in comparison to other dopamine agonists (bromocriptine and lisuride) which stimulate only D2 receptors. PMID- 8708505 TI - Combined chemotherapy trials with regimens containing ofloxacin and rifampicin for multibacillary leprosy patients. AB - The results of combined chemotherapy trials with regimens containing ofloxacin and rifampicin for the treatment of 60 multibacillary leprosy cases from January 1989 to September 1995 was reported. Clinical improvement was achieved by all regimens from the end of the first month. Most patients continued to improve for 3 years. Bacterial indices were gradually reduced during the treatment. Patients on regimens containing rifampicin were clear of M. leprae at the end of the 5th year. The complications and side-effects of ofloxacin and rifampicin were trivial and both drugs were well tolerated. Ofloxacin added to the current WHO recommended M.B.-MDT regimen may shorten the duration of treatment. Ofloxacin may be an alternative in suspected/proven rifampicin resistant cases or rifampicin contraindicated. PMID- 8708506 TI - Effect of salbutamol on oxygen saturation in COPD. AB - The effect of nebulized salbutamol on oxygen saturation in COPD was evaluated in this study. Twenty patients aged 56-75 years with stable COPD were recruited to the study. All patients received 5 mg of salbutamol nebulizer driven by oxygen at 8 L/min. The SaO2 was then measured continuously and readings were taken before and every minute after salbutamol inhalation by face mask. Following treatment, mean SaO2 was significantly lower than preinhalation values. The desaturation occurred from 3 minutes and reached the maximum desaturation at 15 minutes postinhalation. At 30 minutes, SaO2 had returned to baseline values. The findings suggest that salbutamol nebulization induced prolonged desaturation in COPD. PMID- 8708507 TI - Surfactant treatment in the neonate with severe respiratory distress syndrome. AB - Eighteen preterm infants severely ill with respiratory distress syndrome who required assisted ventilaton were given modified natural surfactant (Survanta) endotracheally. They had a mean +/- SEM gestational age of 31.2 +/- 0.4 weeks (range 28-34) and a mean +/- SEM birthweight of 1562 +/- 71 g (range 1160-2010). Average time of initial surfactant administration was 15 +/- 1.7 hour (range 5 24). No significant side effects were found during surfactant instillation. Post surfactant, the air entry was improved, oxygenation and arterial/alveolar gradients increased, and the levels of inspired oxygen could be reduced. Some of the radiological abnormalities were resolved. In 13 infants, patent ductus arteriosus became clinically evident, seven of whom received Indomethacin. There were 4 cases of pulmonary air leak, 5 cases of pulmonary hemorrhage and 8 cases of bronchopulmonary dysplasia. Four infants expired, two were due to severe asphyxia/shock and two died of unrelated causes. Among the 14 survivors who came for follow-up, two cases of retinopathy of prematurity had gradually regressed, one had cerebral palsy and delayed development. Surfactant rescue therapy is a supplemental beneficial treatment for severe respiratory distress syndrome while newborn intensive care concept is necessary for efficient diagnosis and treatment of RDS. PMID- 8708508 TI - Photodynamic therapy in the treatment of head and neck cancers: a two-year experience. AB - This report describes the effectiveness of photodynamic therapy (PDT) in treating patients with widespread superficial premalignant and malignant lesions, primary and second primary, nonresponsive residual or recurrent cancers in head and neck. Twelve patients whose disease was at T1 stage responded completely. Severe dysplasia and field cancerous lesions involving a large area of oral mucosa (3 cases) also yielded excellent results. Average disease-free period was 13.6 months. The longest survival period was 2 years. All patients as mentioned are still alive without any relapse or recurrence. Combined PDT with other treatment was required to control T2-3 carcinomas. PDT adjunct, however, permitted the prescription of conventional treatment, i.e. radiotherapy or surgery, in a less morbid manner. No unacceptable treatment complications and skin photosensitivity were observed in this study. This indicated the potential role of PDT in the management of long-standing problems in head and neck cancers. PMID- 8708509 TI - Blood glucose concentration in pediatric outpatient surgery. AB - Blood glucose concentration was measured in 84 pediatric patients who were scheduled for outpatient surgery at Chulalongkorn Hospital. They were allocated into 3 groups according to their ages, group 1:less than 1 year of age, group 2:1 to 5 years of age and group 3:over 5 years. The fasting times were approximately 8-12 hours. All patients received standard general anesthesia under mask. No glucose solution was given during operation. Preoperative mean blood glucose were 91.09 +/- 17.34, 89.55 +/- 18.69 and 82.14 +/- 16.14 mg/dl in group 1, 2 and 3 while the postoperative mean glucose values were 129.07 +/- 37.90, 115.62 +/- 29.63 and 111.53 +/- 23.07 mg/dl respectively. The difference between pre- and post-operative values were statistically significant difference (P < 0.01). None of the children had hypoglycemia even when fasting longer than expected. Increased postoperative glucose values may be due to stress response from surgery and anesthesia. We would suggest that the parents give the fluid to their children according to our instructions in order to prevent dehydration and hypoglycemia especially in small infants. PMID- 8708510 TI - Paclitaxel in patients with platinum-resistant ovarian cancer: a selected review of literature and clinical experience. AB - Single-agent paclitaxel infused over 24 hours produces response rates of 10 per cent to 20 per cent and 48 per cent respectively, for doses of 135 mg/m2 and 250 mg/m2. This suggests a dose response relationship. However, only a randomized trial comparing the 135 mg/m2 and 250 mg/m2 doses can confirm this result. Unfortunately, the median survival is comparable despite the difference in response rates. This may be secondary to a low CR noted at all doses. The apparent lack of benefit in terms of increased survival for the high dose group, with its attendant increase in incidence of toxic effects and cost (owing to both the paclitaxel and the G-CSF), suggest that the role of higher doses remains to be proven. It may be most useful in alleviating the severe cancer induced symptoms of some patients with advanced platinum resistant ovarian cancer(19). Despite the extensive international experience with paclitaxel in the ovarian cancer clinic its role still needs to be better defined both for salvage therapy and in combination with platinum as a front-line treatment. PMID- 8708511 TI - CD4 + T counts with a course of antituberculous therapy in healthy and HIV infected patients. AB - A prospective study of CD4 + T lymphocytes with a course of treatment on 20 HIV infected patients with active tuberculosis receiving antituberculous drugs was compared with 30 non HIV-infected patients with active tuberculosis. The CD4 + T cell counts among non HIV-infected patients were 510 +/- 409 and increased to 634 +/- 382 and 867 +/- 248 at the third and sixth month of therapy. Among HIV infected patients, the CD4 + T cell counts were 64 +/- 42 and decreased to 40 +/- 23 and 35 +/- 16 in the third and sixth month of therapy (p < 0.05). PMID- 8708512 TI - Spontaneous abortion rate. AB - OBJECTIVE: To study the incidence of spontaneous abortion and relationship between incidence and maternal age profession, education level and income. STUDY DESIGN: Prospective cohort study. SETTING: Department of Obstetrics and Gynecology, Faculty of Medicine, Ramathibodi Hospital, Mahidol University, Bangkok 10400, Thailand. SUBJECTS: 999 patients registered at antenatal care unit, Department of Obstetrics and Gynecology from July 1, 1992. MAIN OUTCOME: Number of spontaneous abortion measures. RESULTS: From 999 patients, 69 were excluded from the study (8 had induced abortion, 61 could not be contacted). Most of patients aged 20-34 (85.5%) with the mean of 27 years. Spontaneous abortion occurred in 64 patients, incidence of 6.9 per cent. The analysis of data showed no relationship between the incidence of spontaneous abortion and age, profession, education level and income. CONCLUSION: Compared with previous studies, the incidence of spontaneous abortion is this study was quite low. With regard to the relationship of socioeconomic, further study with larger population should be performed to draw suitable conclusion. PMID- 8708513 TI - Nutrition knowledge of adolescents in a family setting in Geelong, Australia. AB - A family study of nutrition knowledge was carried out in 213 families in the city of Geelong, Victoria, Australia. On average, mothers had the highest number of correct answers [mean 8.0 (S.D. 1.4)] followed by fathers [mean 7.8 (S.D. 1.5)], adolescent girls [mean 6.4 (S.D. 1.8)] and adolescent boys [mean 6.2 (S.D. 1.5)]. A highly significant difference, in the score obtained, was found between parents and adolescents (p < 0.001). On average, the percentage of 'do not know' answers was twice as high among adolescents (26%) as among parents (11%). Negative correlations which ranged from -0.10 for girls to -0.23 for fathers were found between the knowledge score obtained by the subjects and socioeconomic status. Parents' knowledge score was independent of their educational level (p > 0.05). Mothers with TAFE (Technical And Further Education) qualifications obtained the highest score (8.7). PMID- 8708514 TI - Pheochromocytoma in pregnancy: a case report. AB - A 25-year-old woman, primigravida, developed the clinical picture of sporadic hypertension (blood pressure 120-230/70-130 mmHg), palpitations, flushing and sweating attacks at 34 weeks' gestation. A 24-hour urine collection revealed catechol levels consistent with a diagnosis of pheochromocytoma. The ultrasound and MRI showed a well-defined, rounded soft-tissue mass at the left suprarenal region. The hypertension was controlled with alpha-blockade. Antepartum fetal testing was normal. The patient underwent cesarean section at 38+1 weeks' gestation and delivered a 2,930-grams male with Apgar scores of 9 and 10 at one and five minutes. Left adrenalectomy was performed two weeks later. The pathologic diagnosis was pheochromocytoma. PMID- 8708515 TI - Burkholder meningitis. AB - We report two cases of Burkholder meningitis. Case 1 presented with subacute lymphocytic meningitis, low sugar profile in CSF and positive culture for Burkholder. Case 2 presented with purulent meningitis that emerged during treatment of cryptococcal meningitis which, to our knowledge, has not been reported. Thus, Burkholder should be considered as one of the etiologic agents in this clinical setting. PMID- 8708516 TI - Resolution of dilated cardiomyopathy after radio-frequency catheter ablation of atrial tachycardia. AB - The dilated cardiomyopathies are characterized by an increase in left ventricular internal dimensions without an appropriate increase in ventricular wall thickness. Sustained tachyarrhythmia, either supraventricular or ventricular, may be associated with this condition and may improve after termination of the tachycardia. We report a case of secondary dilated cardiomyopathy caused by incessant atrial tachycardia who underwent successful radiofrequency catheter ablation and left ventricle returned to normal size and function in six months post ablation. PMID- 8708517 TI - The effectiveness of hormone in relieving menopausal symptoms. AB - To assess the effectiveness of hormone replacement in relieving menopausal symptoms, 75 natural menopausal women suffering from these symptoms were recruited for this double-blind, placebo controlled study. These women were randomized into 2 groups. In the first eight weeks, the first group received a sequential regimen of Estradiol valerate 2 mg/day for 21 days and Norgestrel 0.5 mg/day from day 12 to 21, with seven days of drug-free interval, for two cycles. In the last eight weeks, placebo was given in the same fashion. In the second group, placebo was given in the first eight weeks, then hormone in the last eight weeks. The menopausal scores were recorded by the patients at weeks 0, 4, 8, 12 and 16. Sixty women completed the 16 week study, of which there were 30 women in each group. The mean menopausal score by eight weeks decreased significantly in the first group compared to the pretreatment value (The scores at week 0 and 8 = 20.03 and 12.63 respectively, p < 0.05). However, there was no significant change of the score in the second group. (The scores at week 0 and 8 = 21.20 and 19.43 respectively, p > 0.05). After switching from hormone to placebo in the first group, there was no significant change in mean menopausal score (The scores at week 8 and 16 = 12.63 and 13.73 respectively, p > 0.05). Nevertheless, when switching from placebo to hormone in the second group, the score decreased significantly. (The scores at week 8 and 16 = 19.43 and 13.20 respectively, p < 0.05). In conclusion, this study demonstrated the beneficial effects of hormone replacement in relieving menopausal symptoms. PMID- 8708518 TI - Dementia in Thai patients with Parkinson's disease: analysis of 132 patients. AB - A cross-sectional design study was done at Siriraj Hospital from 1990 to 1994 to determine the prevalence of dementia in Thai patients with Parkinson's disease. There were 81 males and 51 females with a male to female sex ratio of 1.59:1. One hundred and thirty two patients with Parkinson's disease were enrolled in this study. The mean duration of illness was 3.5 (S.D. 3.53) years. All patients were screened for depression by using the Thai Geriatric Depression Scale and patients with Alzheimer's disease or stroke were excluded from the study. The Thai Mental State Examination (TMSE) is a standard test used in this study to identify dementia in Thai patients with Parkinson's disease. Thirty four patients (25.76 per cent) scored TMSE below 23 points (cut-off point for dementia) and were considered as having dementia. Twenty four patients (18.18 per cent) scored below 20 points and were considered as having severe dementia, 12 patients (9.09 per cent) scored between 20-22 points (classified as mild to moderate dementia). Thirty seven patients (28.03 per cent) were in the borderline group as they had TMSE scores between 23-25 points. Fifty nine patients (44.70 per cent) were determined as nondemented as their TMSE scores were over 25 points. The items of cognitive function tests in TMSE which were severely impaired in the demented group were recall, attention and calculation whereas registration was relatively unaffected. Orientation and language were moderately impaired in the demented group. Dementia in Thai patients with Parkinson's disease is a significant problem for long term care of patients because of its high prevalence (one-fourth of all patients) and nearly one-fifth had severe dementia. Prompt recognition and proper management of dementia are required for family members and medical personnel to handle patients with Parkinson's disease. PMID- 8708519 TI - Effect of isoniazid prophylaxis on incidence of active tuberculosis among Thai HIV-infected individuals. AB - A prospective comparative study was conducted to determine the effect of isoniazid prophylaxis on the incidence of active tuberculosis among Thai HIV infected patients for 1 year. Among those 36 HIV-infected patients without prophylaxis, the incidence of active tuberculosis was 2.7 per cent while in 10 HIV-infected patients with isoniazid prophylaxis, there was no incidence of active tuberculosis during the first year. PMID- 8708520 TI - Hyperthermia conjunction with radiation therapy in breast cancer with chest wall recurrence: a preliminary report. AB - 1. Hyperthermia is a very effective and valuable tool which produces a high degree of response in chest wall recurrent breast cancer, especially in cases, where curative doses of radiation could not be achieved, due to previous radiation treatment. 2. Hyperthermia once a week is sufficient to produce the enhancement of radiation neither increasing the burden to the patient nor increasing the workload to the department. 3. The effective radiation dose which causes no severe complication is 50 Gy in a small tumor and 60 Gy in a large tumor. 4. In small satellite nodules, whole chest wall irradiation should be considered to prevent recurrence in the adjacent area. 5. An air cooling system is effective and suitable for an ulcerative lesion. Further randomized study for long term local control and survival should be explored. PMID- 8708521 TI - Retinal detachment with macular hole. AB - Retinal detachment with macular hole was treated by different methods. Intravitreal SF6 injection was helpful to seal the macular hole and reattach the retina. In cases of retinal detachment with macular hole and peripheral retinal tear, the retinal reattachment could be achieved by scleral buckling procedure. When vitreous traction adjacent to the macular hole or proliferative vitreoretinopathy was present, vitrectomy combined with internal temponade by SF6 gas injection was indicated. Failure of the operation was caused by inadequate removal of the vitreous traction and post-operative vitreoretinopathy. In a successful operation, visual acuity of 6/60 or better was found in eight of thirty-one patients. (25.8%) PMID- 8708522 TI - Mercury content in the gallstones and bile of Thai people (Chiang Mai and Bangkok) and Japanese. AB - Mercury contents in the gallstones and bile of patients living in Thailand (Chiang Mai and Bangkok) and Japan were studied by neutron activation analysis. The correlation between contents in the gallstones and bile, and the amount of mercury in the diet and environment was investigated by comparing the contents in patients living in three different areas. The content in the bile of the Japanese patients was significantly greater than that of the Thai patients. The distribution of mercury contents in the pigment and rare stones of patients living in Bangkok indicated a bipolarity. However, the distribution of the contents in cholesterol, pigment, and rare stones of Japanese patients did not indicate a bipolarity. Bipolarity would indicate the amount of mercury in dietary foods, and the pollution level of mercury. The mercury contents in the gallstones and bile would reflect the amount of mercury in the diet and environment. PMID- 8708523 TI - Pharmacokinetics of oral lidocaine and nifedipine in patients with liver cirrhosis. AB - The pharmacokinetics of oral lidocaine and nifedipine and hemodynamic effects of nifedipine were studied in 10 cirrhotic patients and 10 healthy volunteers. In a randomized two-way crossover design, each subject received 50 mg of lidocaine solution and 10 mg capsule of nifedipine with one-week washout period. After oral lidocaine, cirrhotic patients has a longer time to peak concentration (Tmax) and elimination half-life (t1/2), and a higher area under the curve (AUC). There were no significant differences in peak plasma concentration (Cmax) and elimination rate constant (K(el)) in the two groups. After oral nifedipine, cirrhotic patients had a longer elimination t1/2, lower K(el) and higher AUC. At peak concentration in cirrhotic patients, there was more decrease in the systolic blood pressure and less increase in heart rate. Although large interindividual variability existed in this study, pharmacokinetic parameters were considerably altered in cirrhotic patients. PMID- 8708524 TI - Percutaneous coil embolization and balloon-expandable stenting; new treatments in intractable cardiac failure after Fontan procedure: a case report. AB - Successful percutaneous coil embolization of aortopulmonary collaterals and balloon-expandable stent implantation of left pulmonary artery stenosis in a child with hypoplastic right heart syndrome were reported, Presently, interventional cardiac catheterization is a very useful treatment strategy in congenital heart diseases unamenable to or inaccessible for surgery. PMID- 8708525 TI - Serum transcobalamin II level in glucose-6-phosphate dehydrogenase deficient subjects with typhoid fever. AB - Transcobalamin II (TCII) is the vitamin B12 binding protein which is responsible for delivery of this vitamin to the tissues. High values for serum TCII have been reported in many clinical conditions. This paper describes the elevated serum TCII levels in three G-6-PD deficient patients with typhoid fever. They had severe hemolysis with hemoglobinuria associated with slight liver dysfunctions but without obvious increased serum creatinine and BUN concentrations. A remarkable increase in serum TCII level was observed during active hemolysis and decreased to the normal level within 2-3 days after hemolysis ceased. The mechanism of increased serum TCII during hemolysis is probably due to hemoglobinuria secondary to excessive hemolysis. As Hb is known to be efficiently reabsorbed by the proximal tubule cells and can competitively inhibit the tubular uptake of TCII-B12. It is possible that excess Hb interferes with TCII uptake and degradation at renal tubular cells. Therefore, the circulating TCII survival is prolonged resulting in the elevated TCII level. Furthermore, lysosomal degradation of newly synthesized TCII is a normal process that regulates the TCII secretion. Therefore, a reduced lysosome-mediated uptake of TCII-B12 by renal tubular cell may stimulate the TCII secretion as has been shown experimentally in vitro. PMID- 8708526 TI - Danazol for thrombocytopenia in pregnancy with underlying systemic lupus erythematosus. AB - A case of full blown systemic lupus erythematosus (SLE) was treated with steroid. The patient was in remission with low dose of prednisolone for a year. When she became pregnant, there was no relapse of SLE activities in any organ. However, she developed gum bleeding and petechiae due to thrombocytopenia in the second half of the pregnancy. With a normal amount of megakaryocyte in the bone marrow, it was supposed to be due to SLE which no longer responded to even a full dose of steroid. Therefore, danazol 600 mg/day was given orally and she made a complete recovery within one week. She delivered vaginally a normal female newborn without fetal thrombocytopenia or bleeding. PMID- 8708527 TI - Salmonella lung abscess and bacteraemia in an AIDS patient. AB - This report describes a 28-year-old, HIV-infected man presenting with subacute onset of pyrexia, cough, dyspnoea and pleuritic pain. Chest radiograph showed bilateral multiple cavitary lesions. The diagnosis of salmonellosis was secured by isolation of salmonella typhimurium in blood, as well as in sputum. Therapy with sequential ceftriaxone/ciprofloxacin led to satisfactory improvement symptomatically and radiologically. The present report serves to heighten the awareness of AIDS-associated salmonella bacteremia and lung abscesses. PMID- 8708528 TI - Bacillary angiomatosis and mycobacterium infection coexisting in a cutaneous lesion in a patient with AIDS. AB - Bacillary angiomatosis is a recently recognized bacterial infectious disease. It mainly affects patients with acquired immunodeficiency syndrome. The presence of coexistent infections of more than one pathologic process in skin lesions in patients with AIDS has been demonstrated. We report a patient with AIDS in whom both bacillary angiomatosis and mycobacterium infection were documented within the same cutaneous lesion. PMID- 8708529 TI - The neonatal period: a critical interval in male primate development. AB - It appears from the evidence documented in this Commentary that the neonatal period is another critical stage in the process of sexual, behavioural and immune system development and maturation in primates. Interference with normal brain pituitary-gonadal function during this period (e.g. with a GnRH analogue in monkeys) appears to impact adversely on subsequent reproductive, immunological and behavioural function. These data further emphasize the importance of fully understanding the regulatory mechanisms that govern neonatal gonadal function in the primate, if we are to eliminate, control or minimize the potential risk resulting from its disruption in humans. Given the recent evidence that the reproductive potential of the human male has declined rather dramatically over the last 50 years, and that clinical conditions associated with abnormal testicular function are on the rise, continued investigation in this area would appear to be imperative. PMID- 8708530 TI - Gaseous transmitters as new agents in neuroendocrine regulation. PMID- 8708531 TI - Expression of IGF-I and IGF-binding protein genes in cirrhotic liver. AB - The liver plays a central role in the IGF-I axis producing the majority of circulating hormone and some of its binding proteins (IGFBPs). Cirrhosis of the liver is characterised by changes in IGF-I and IGFBPs associated with liver fibrosis and regeneration. We have studied steady state levels of mRNA for the genes in the IGF-I axis in normal and cirrhotic human liver, localised the most highly expressed gene, IGFBP-1, and measured circulating IGFBP-3 by radioimmunoassay (RIA), IGFBP-2 and IGFBP-3 by Western ligand blot (WLB), and protease activity for IGFBP-3 in cirrhotic patients. Messenger RNA for IGF-I, IGFBP-1, IGFBP-2, and IGFBP-3 was detectable by Northern blotting in normal and cirrhotic liver although there was considerable variation in expression. IGFBP-2 and IGFBP-3 tended to be more highly expressed in cirrhotic liver and IGFBP-1 was more highly expressed in normal liver, although there were no significant differences. In normal liver, in situ hybridisation localised IGFBP-1 to hepatocytes. In cirrhotic liver the regenerating nodules showed expression of IGFBP-1 while there was none in fibrotic tissue. Circulating IGFBP-3 levels were low as measured by RIA and WLB but protease activity was only found in one patient. IGFBP-2 levels, assessed by WLB, were similar to the normal serum pool. Our data show that key mRNAs involved in the IGF-I axis continue to be expressed in cirrhotic liver despite end stage liver disease. The low levels of IGFBP-3 do not appear to be due to reduced gene transcription or increased protease activity. PMID- 8708532 TI - Localization and distribution of angiotensin II in the rat epididymis. AB - The localization and distribution of angiotensin II (Ang II) in the rat epididymis was studied using immunohistochemical and RIA techniques. The immunohistochemical results showed that Ang II-like immunoreactivity progressively increased along the length of the rat epididymis (cauda > corpus >> caput) and was predominately localized in the basal region of the epididymal epithelium. Occasionally, immunostaining of lighter intensity was also found in the apical region. The concentration of Ang II in cultured rat cauda epididymal epithelial cells was further measured by RIA. In addition to that found in cultured epithelial cells, Ang II activity was also detected in the culture medium, suggesting a secretory role of the epithelium. These findings suggest that Ang II could be derived locally from epididymal epithelium and that it could play a role in local regulation of epithelial transport and, possibly, in the maintenance of sperm function as well, by exerting its paracrine and/or autocrine effect in various regions of the epididymis. PMID- 8708533 TI - The effects of octreotide on GH receptor and IGF-I expression in the GH-deficient rat. AB - Somatostatin has been suggested to influence the somatotrophic axis outside the central nervous system, in reducing GH-induced IGF-I mRNA and IGF-I generation. This study aimed to determine whether such effects were mediated via the GH receptor (GHR). GH-deficient dwarf rats aged 45-47 days (n = 8 per group) received twice daily subcutaneous injections of octreotide (1 mg/kg) (group O), saline (group S), octreotide (1 mg/kg) plus bovine GH (0.25 mg/kg) (group OG), or bovine GH (0.25 mg/kg) plus saline (group G) for 10 days. Octreotide-treated animals had less weight gain compared with saline-treated animals, but not when GH cotreated (group OG vs G). Octreotide had an overall effect on decreasing length gain (P < 0.01). Serum IGF-I (ng/ml) was reduced by octreotide (group O 171 +/- 11, group S 239 +/- 20, P < 0.01; group OG 283 +/- 30, group G 362 +/- 10, P < 0.001), as was serum insulin (P < 0.001). A significant decrease in hepatic and muscle IGF-I mRNA expression was found as expected, yet this was not associated with decreased hepatic GHR expression. Rather, an increase in hepatic 125I-bovine GH specific binding was observed (P < 0.001) and, in GH-cotreated animals (OG), hepatic GHR and GH binding protein (GHBP) mRNA expression were also increased by octreotide by approximately 40%. In muscle, octreotide was associated with an approximately 30% decrease in GHBP mRNA and no effect on GHR mRNA. This study suggests that the suppressive effects of octreotide on IGF-I metabolism, at least in liver, are not mediated via down-regulation of GHR expression, but more likely by direct effects on IGF-I expression. PMID- 8708534 TI - The role of pregnenolone-metabolizing enzymes in the regulation of oestradiol biosynthesis during development of the first wave dominant follicle in the cow. AB - During the luteal phase in the cow, a first-wave dominant follicle grows to reach ovulatory size, but then ceases to grow, becomes no longer dominant and enters a phase of slow regression. During this growth transition, the concentration of oestradiol has been shown to decrease in follicular fluid. The objective of this study was to determine if follicular fluid oestradiol concentrations are regulated by the activity of three major steroidogenic enzymes, namely P450 aromatase (P450-arom), 3 beta-hydroxysteroid dehydrogenase/delta 5-delta 4 isomerase (3 beta-HSD) and 17 alpha-hydroxylase C-17,20 lyase cytochrome P450 enzyme (P450-17 alpha) measured in granulosa and theca cells isolated from individual first-wave dominant follicles. Follicle growth and state of dominance was assessed by ultrasonography and follicles were classified as growing-dominant (GD, n = 6), non-growing-dominant (NGD, n = 8) or non-growing-non-dominant (NGD, n = 6). Mean follicular fluid concentrations of oestradiol were higher in GD than in NGD or NGND follicles (511 +/- 98 versus 136 +/- 16 and 20 +/- 11 nmol/l respectively). Oestradiol was not correlated with P450-arom in any of the three groups. In GD follicles, oestradiol was positively correlated with pregnenolone concentration but neither was correlated with granulosa or theca 3 beta-HSD activity or with theca P450-17 alpha activity. In NGD follicles, oestradiol was negatively correlated with theca 3 beta-HSD activity and pregnenolone was negatively correlated with granulosa 3 beta-HSD activity. In NGND follicles, oestradiol was positively correlated, and pregnenolone was negatively correlated with theca 3 beta-HSD and P450-17 alpha activities. These studies demonstrated that pregnenolone supply is the principal regulating factor of oestradiol output during follicle dominance and during the loss of dominance but that the levels of P450-17 alpha and 3 beta-HSD activity become rate-limiting when the follicle is no longer dominant. PMID- 8708535 TI - Immunohistochemical localization of the glucocorticoid receptor in human fetal membranes and decidua at term and preterm delivery. AB - The human fetal membranes and decidua may be important in the onset and/or progression of human labor by providing prostaglandins for this process. Glucocorticoids have been implicated in the regulation of prostaglandin production by these tissues but to date there is no direct evidence for glucocorticoid receptors (GRs) being present in human intrauterine tissues. The purpose of the present study was to determine, using immunohistochemistry, whether the human fetal membranes and decidua contained GRs; to determine the localization of receptors to the cytoplasm or nuclei, and to examine the content and distribution of the GRs in tissues obtained during pregnancy following preterm labor (< 37 weeks) and at term prior to and following term labor. Term tissues were obtained prior to labor by elective Caesarean section (n = 9) or following vaginal delivery (n = 9). Tissues from 14 patients who delivered preterm but with no clinical evidence of infection were also examined. Cryostat sections were thaw-mounted onto microscope slides. The immunoreactive GRs were visualized with an Elite Vectastain ABC Kit using a polyclonal antibody prepared against a synthetic peptide corresponding to amino acids 346-367 of the human GR. At term, nuclear GRs were found in amnion epithelial cells, mesenchyme and the chorion laeve. GRs were present, but were less defined, in the decidua. A similar distribution was found in the preterm tissues. However, nuclear staining in the amnion epithelial cells, mesenchymal cells, chorion and decidua was more pronounced in tissues obtained following preterm labor. This study provides direct evidence for the presence of GRs in human fetal membranes and decidua, and suggests the possible importance of multiple cell types in the action of glucocorticoids in these tissues. PMID- 8708536 TI - Expression of granulocyte-colony stimulating factor and its receptor is regulated during the development of the human placenta. AB - The development of the placenta is dependent upon the regulated proliferation, invasion and differentiation of trophoblast. Expression of cytokines at the feto maternal interface suggests that these molecules may participate in placentation. The expression of granulocyte-colony stimulating factor (G-CSF) and G-CSF receptor (G-CSFR) during the development of the human placenta was studied by immunohistochemistry using an anti-G-CSF monoclonal antibody (mAb) and two novel anti-G-CSFR mAbs. G-CSF was present in the stroma of fetal chorionic villi and maternal decidual tissues throughout pregnancy. G-CSFR was detected at high levels in fetal first and third, but not second trimester placental tissues. Staining for G-CSFR was undetectable in maternal decidual tissue from all gestational stages. In first trimester tissues, staining for placental G-CSFR was strongest in differentiated syncytiotrophoblast and invasive, extravillous cytotrophoblast, and weak staining was evident in undifferentiated cytotrophoblast. Immunohistochemical data suggesting temporal regulation of G CSFR were corroborated by Western blotting and amplification by reverse transcription and PCR of G-CSFR mRNA. These data suggested that expression of G CSFR in the human placenta is regulated both temporally and spatially, and that placental G-CSF is involved in paracrine regulation, and indicate a role for G CSF and G-CSFR in trophoblast growth or function during placentation. PMID- 8708537 TI - Induction and involvement of endogenous IGF-I in pancreas regeneration after partial pancreatectomy in the dog. AB - To elucidate whether and how IGF-I is involved in the regeneration of the pancreas after partial pancreatectomy, IGF-I mRNA expression, IGF-I protein synthesis, ornithine decarboxylase (ODC) activity and DNA replication in the remnant pancreas were determined in the dog. After pancreatectomy, IGF-I mRNA expression was remarkably enhanced in the remnant pancreas, showing the maximal value at post-operative day (POD) 1. Subsequently, IGF-I synthesis in the tissue was significantly stimulated at POD 2, and its maximal concentration was observed at POD 3. Following IGF-I synthesis, ODC activity was induced and its maximal activity was also obtained at POD 3. Finally, DNA replication was induced in the remnant pancreas, and its maximal level was observed at POD 5. These responses in the remnant pancreatic tissue to partial pancreatectomy were greatly enhanced as the resection rate was increased up to 95%. Positive correlations were observed between IGF-I concentrations in the remnant pancreas and the activities of ODC and DNA synthesis in the tissue after 95% pancreatectomy. These results suggest that the gene expression of IGF-I is rapidly induced in the remnant pancreas after partial pancreatectomy, and subsequently synthesized endogenous IGF-I peptides may stimulate ODC and other cell growth-related activities in the tissue in paracrine and/or autocrine manners eventually to induce DNA replication and tissue regeneration. PMID- 8708538 TI - Parathyroid hormone induces superoxide anion burst in the osteoclast: evidence for the direct instantaneous activation of the osteoclast by the hormone. AB - We have shown that superoxide anion (O2-) production by the osteoclast can be used as an index of the osteoclast activity since the agents that inhibit and stimulate the osteoclast also diminish and stimulate O2- production respectively. Therefore, we have investigated the mechanism of parathyroid hormone (PTH) mediated stimulation of osteoclast function in terms of its effect on O2- generation. The determination of O2- generation was carried out by employing cytochrome c immobilised on a surface-modified gold electrode. The basal level of free radical production by the osteoblast-like cells (ROS 17/2.8) was 10(4)-fold lower than by osteoclasts cultured on bone. PTH had no acute effect on free radical production by the osteoblasts. The exposure of the osteoclasts cultured on bone to PTH led to a dramatic and immediate stimulation of O2- generation which was unaffected by the presence of ROS 17/2.8 cells. The osteoclasts co cultured with ROS 17/2.8 cells and exposed to PTH for 3 h were also found to produce greater stimulation of O2- than the osteoclasts exposed to PTH alone. A competitive leukotriene D4 antagonist REV 5901, which also inhibits 5 lipoxygenase, did not block O2- generation by osteoclasts cultured alone or in the presence of osteoblasts. Therefore, we conclude that PTH directly stimulates osteoclasts to produce O2-; this may be the main mode of activation of the osteoclasts, although an osteoblast-mediated effect of the hormone cannot be ruled out. PMID- 8708539 TI - The expression of transforming growth factor-beta by cultured chick growth plate chondrocytes: differential regulation by 1,25-dihydroxyvitamin D3. AB - 1,25-Dihydroxyvitamin D3 (1,25(OH)2D3) and transforming growth factor-beta (TGF beta) are both important regulators of chondrocyte growth and differentiation. We report here that 1,25(OH)2D3 differentially regulates the expression of the genes for TGF-beta 1 to -beta 3 and the secretion of the corresponding proteins in cultured chick chondrocytes. Confluent growth plate chondrocytes were serum deprived and cultured in varying concentrations of 1,25(OH)2D3. Cells were assayed for TGF-beta mRNA and conditioned medium was assayed for TGF-beta activity and isoform composition. Active TGF-beta was only detected in 10(-8) M 1,25(OH)2D3-treated cultures (8.37 ng active TGF-beta/mg protein). There was a significant decrease in total (latent-active) TGF-beta activity in conditioned medium of 10(-12) M (23.4%; P < 0.05) and 10(-10) M (20.7%; P < 0.05) 1,25(OH)2D3 treated cultures but 10(-8) M 1,25(OH)2D3 significantly increased (30.9%; P < 0.01) TGF-beta activity. The amounts of TGF-beta 1, -beta 2 and -beta 3 isoforms produced were similar in control, 10(-10) or 10(-12) M 1,25(OH)2D3-treated cultures but the conditioned medium of 10(-8) M 1,25(OH)2D3-treated cultures contained significantly higher amounts of all three isoforms. Quantification of TGF-beta mRNA demonstrated differential control of TGF-beta gene expression with TGF-beta 1 and -beta 3 mRNA levels reduced by all concentrations of 1,25(OH)2D3 examined (10(-8), 10(-10) and 10(-12) M) whilst TGF-beta 2 mRNA concentrations were elevated. Our results indicated that 1,25(OH)2D3 regulates chick growth plate chondrocyte TGF-beta secretion and mRNA expression in a concentration dependent and isoform-specific manner. This interaction may be important in the regulation of chondrocyte metabolism and endochondral bone growth. PMID- 8708540 TI - Pituitary adenylate cyclase-activating peptide stimulates cyclic AMP accumulation in UMR 106 osteoblast-like cells. AB - Pituitary adenylate cyclase-activating peptide (PACAP) and vasoactive intestinal peptide (VIP) share 68% homology and function as neurotransmitters or neuroendocrine factors. Although VIP immunoreactivity has been detected in bone cells, the presence of PACAP or PACAP receptors in bone has not been determined. In this study, we investigated the role of PACAP and VIP in regulating cAMP accumulation in the UMR 106 osteoblast-like tumor cell line. PACAP 27 (10(-9) to 3 x 10(-7) M), PACAP 38 (10(-9) to 3 x 10(-7) M) and VIP (10(-8) to 10(-6) M) stimulated cAMP accumulation up to eightfold. PACAP 27 was slightly more potent than PACAP 38, and both were tenfold more potent than VIP. Both PACAP- and VIP stimulated cAMP accumulation were potentiated by 4 beta-phorbol 12-myristate 13 acetate, an activator of protein kinase C. Two PACAP antagonists, PACAP 6-27 (3 x 10(-6) M) and PACAP 6-38 (3 x 10(-6) M), blocked PACAP- and VIP-stimulated cAMP accumulation. Two VIP antagonists ([Lys1,Pro2,5,Arg3,4,Tyr6]-VIP, and [4 Cl-D Phe6,Leu17]-VIP) did not reduce the PACAP- or VIP-stimulated cAMP accumulation. Pretreatment with PACAP 27, PACAP 38 or VIP equally blocked PACAP- and VIP stimulated cAMP accumulation. These results suggest that PACAP is a more potent stimulator of cAMP accumulation than VIP in UMR 106 cells. PACAP and VIP may share a role in the paracrine or neuroendocrine regulation of bone metabolism. PMID- 8708541 TI - Insulin-like growth factor-binding protein-6 produced by human PC-3 prostate cancer cells: isolation, characterization and its biological action. AB - The PC-3 human prostatic carcinoma cell line has been extensively used as a model for studies on the regulation of prostate tumor cell proliferation. Because of the importance of IGF-binding proteins (IGFBPs) in the control of IGF activities that regulate cell proliferation in normal and malignant cell types, we undertook studies to characterize the IGFBPs produced by PC-3 prostate tumor cells in culture. We previously found, using an IGF-I affinity column for purification and a polyethylene glycol (PEG) precipitation assay for IGFBP detection, that PC-3 cells in culture produced a single predominant IGFBP, IGFBP-4, which inhibits IGF activities. We now present evidence that PC-3 cells also produce IGFBP-6 in abundant quantity; in the previous study this was apparently not detected in the IGF-I-bound fraction with the PEG precipitation and Western ligand blot assays. In the current study, IGF-II affinity purification of IGFBPs produced by PC-3 cells, followed by C8 HPLC reverse-phase chromatography using a shallow acetonitrile gradient, produced two major protein peaks. N-terminal amino acid sequence of peak 1 was identical to that of IGFBP-6 while that of peak 2 was identical to that of IGFBP-4. Characterization of purified IGFBP-6 from PC-3 cells revealed properties which are distinct from other IGFBPs. PEG did not precipitate the complex of 125I-IGF-II/IGFBP-6 while it precipitated the complexes between 125I-IGF-II and other IGFBPs. Indeed, IGFBP-6 decreased the amount of 125I-IGF-II tracer in the PEG precipitate in a dose-dependent manner. Also, the binding of IGFBP-6 with 125I-IGF-II was poor in Western ligand blots compared with other IGFBPs. In studies on IGFBP-6 actions, IGFBP-6 completely inhibited IGF-II-induced [3H]thymidine incorporation in MC3T3-E1 mouse osteoblast cells while it had only minimal inhibitory effects on IGF-I-induced [3H]thymidine incorporation. This differential effect is associated with the fact that IGFBP-6 has greater affinity for IGF-II than IGF-I. The results of this study indicated that (1) Western ligand blotting is not sensitive for identification of IGFBP-6, (2) the unique behavior of IGFBP-6 in the PEG assay system necessitates the use of charcoal adsorption procedure for IGFBP-6 activity detection and (3) PC-3 cells should provide a useful model system for studying regulation of IGFBP-6 expression and the role of IGFBP-6 in modulating IGF actions. PMID- 8708542 TI - Anti-idiotypic antibody as an oestrogen mimetic in vivo: stimulation of creatine kinase specific activity in rat animal models. AB - Previous studies indicated that the anti-idiotypic antibody (clone 1D5) significantly increased the specific activity of creatine kinase (CK) activity in the rat uterus, and in vitro in skeletal cells capable of responding to oestradiol (E2), suggesting that the antibody has oestrogenic-like activity. Moreover, the F(ab')2 dimer of clone 1D5 acted like an antagonist and completely inhibited the increase in CK specific activity by either E2 or clone 1D5 in these skeletal cells. In the present study, we examined the in vivo effects of clone 1D5 and its proteolytic fragment, the F(ab')2 dimer, E2 and dihydrotestosterone (DHT) on CK specific activity in the epiphyseal cartilage, diaphyseal bone, uterus, prostate, thymus and pituitary of immature or gonadectomized female and male rat animal models. In the intact immature animals, clone 1D5 caused an increase in CK in all organs of the female except in the pituitary. In the diaphyseal bone and prostate of male rats there was no stimulation by 1D5. The CK response in the uterus, epiphysis, and diaphysis of immature female rats was dose dependent and was blocked by either the anti-oestrogen tamoxifen or the F(ab')2 dimer of clone 1D5. E2, DHT, as well as clone 1D5, stimulated CK specific activity in both the diaphysis and epiphysis of ovariectomized female and castrated male rats, whereas sex specificity in the CK response was observed also in the uterus and the prostate of gonadectomized animals. Collectively, these results suggest that, as in cell culture, an intact antibody is necessary for the observed stimulation of CK specific activity and the F(ab')2 dimer can act as an antagonist. Furthermore, the observed biological effects of clone 1D5 which are absolutely parallel to E2, imply that the anti-idiotypic antibody is able to penetrate the cell and reach the nuclear oestrogen receptor and transduces a signal to the nucleus, by as yet uncharacterized mechanisms. PMID- 8708543 TI - Gender differences in the associations between cortisol and insulin in healthy subjects. AB - To investigate the role of cortisol in the etiology of insulin resistance in men and women, we examined 218 healthy non-hospitalized elderly, selected from the Rotterdam Study. Free cortisol was assessed by the ratio of fasting serum cortisol over corticosteroid-binding globulin (CBG), and insulin resistance was estimated by the fasting insulin level. CBG was higher in women and decreased with age. In both men and women, the early morning free cortisol level showed no association with age or waist/hip ratio. In men, an inverse association between cortisol and body mass index was observed. In women, higher cortisol levels were associated with increased insulin levels; an increase of 9.7 mU/l insulin per unit cortisol/CBG (S.E. 3.9, P = 0.01). The association did not change after adjustment for age, body mass index or waist/hip ratio. The results of this study in elderly subjects suggest that in women cortisol may be implicated in the age associated insulin resistance. PMID- 8708544 TI - Demonstration of in vivo metabolic effects of 3,5-di-iodothyronine. AB - The objective of the present study was to test in vivo the metabolic effects of 3,5-di-iodothyronine (3,5-T2) in unanesthetized and unrestrained male Sprague Dawley rats. Amino acid and lipid metabolisms were investigated by breath tests using as tracers the 13C-carboxyl-labeled molecules of leucine, alpha ketoisocaproic acid (KIC) and octanoic acid, in four different groups of rats: hypothyroid animals (receiving propylthiouracil (PTU) and iopanoic acid), hypothyroid animals treated with either a daily i.p. injection of 3,5-T2 (25 micrograms/100 g body weight), or tri-iodothyronine (T3) (1 microgram/100 g body weight), and control euthyroid animals receiving equivalent volumes of the vehicle solutions. Energy expenditure was measured by continuous monitoring of O2 consumption and CO2 production in these different groups. Daily energy expenditure was decreased by 30% in PTU-treated rats. The chronic treatments with 3,5-T2 and T3 restored daily energy expenditure to the control level. 13CO2 recovered in breath following the i.v. injection of octanoic acid-[1-13C] was decreased in hypothyroid animals compared with control animals (P < 0.05) and restored to control values by T3 and 3,5-T2 treatments. The 13CO2 recovered in breath after i.v. injection of leucine-[1-13C] was increased in PTU-treated compared with control animals (P < 0.05). Chronic treatment with either 3,5-T2 or T3 restored 13CO2 to control values. Excretion of 13CO2 recovered in breath following the i.v. injection of KIC-[1-13C] was increased in PTU-treated compared with control animals. Chronic treatments with either 3,5-T2 or T3 did not restore KIC decarboxylation. These results suggest that 3,5-T2 exerts metabolic effects on energy expenditure, on both lipid beta-oxidation and leucine metabolism in hypothyroid rats. We conclude that 3,5-T2 is a metabolically active iodothyronine. PMID- 8708545 TI - 1,25-dihydroxyvitamin D3 increases the sensitivity of human renal carcinoma cells to tumor necrosis factor alpha but not to interferon alpha or lymphokine activated killer cells. AB - Renal cell carcinoma is a chemotherapy-resistant tumor which is relatively responsive to immunotherapy. Immunotherapeutic regimes employ interferons or interleukin 2 with or without lymphokine-activated killer cells. Secondary cytokines, induced by interleukin 2 or interferon, may have an important impact on their anti-neoplastic activity. Notable among them is tumor necrosis factor (TNF alpha). We assessed the effect of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) on the susceptibility of the human renal cell carcinoma cell line SK-RC-29 to the cytotoxic and cytostatic actions of TNF alpha, interferon alpha and lymphokine activated killer cells. Using uptake of the vital dye neutral red as an indicator of viable cell number, we found that addition of 1,25(OH)2D3 (100 nM) to TNF alpha (30 ng/ml)-treated cultures resulted in a 2.6 +/- 0.2-fold (mean +/- S.E.) increase in the cytotoxic effect of the cytokine. The potentiating effect of 1,25(OH)2D3 was dose-dependent, and significant at concentrations equal to or higher than 10 nM. Another dihydroxylated vitamin D metabolite, 24,25(OH)2D3, had no effect on TNF alpha action. The cytotoxic effect of TNF alpha increased whereas the potentiation by 1,25(OH)2D3 decreased with cell density in culture. 1,25(OH)2D3, in contrast to its potentiating effect on TNF alpha action, did not modulate the cytostatic effect of interferon alpha or the susceptibility of SK-RC 29 to killing by lymphokine-activated killer cells. The findings reported here may explain some of the in vivo anti-tumor activity of 1,25(OH)2D3 and provide a rationale for the employment of active vitamin D analogs during immune anti cancer therapy. PMID- 8708546 TI - Pup contact induces the expression of long form prolactin receptor mRNA in the brain of female rats: effects of ovariectomy and hypophysectomy on receptor gene expression. AB - Prolactin receptor (PRL-R) mRNA expression levels in the female rat brain (cerebrum) during pup contact stimulation were determined by the reverse transcription-PCR method. The high expression levels of long form PRL-R mRNA found in the brain of lactating rats were markedly reduced by removal of pups, and long form PRL-R mRNA levels were recovered by resumption of pup contact. Interestingly, pup contact stimuli of nulliparous virgin rats also markedly induced long form but not short form PRL-R mRNA expression in the brain in 1.3 days, together with the expression of maternal behaviour. In ovariectomized (OVX) or hypophysectomized (HYPOX) virgin rats, or in OVX plus HYPOX virgin rats, however, brain long form PRL-R mRNA was not significantly induced by pup contact stimuli for as long as 7 days, while maternal behaviour was fully expressed in these rats after 7 days of pup contact. The in situ hybridization experiments revealed that the long form PRL-R mRNA induced in virgin rats in contact with pups or in lactating rats was localized in the epithelial cells of the choroid plexus. No significant increase in mRNA was detected in other regions of the brain, such as the hypothalamus or cortex, in these maternal female rats. These results suggest that pup contact induces the expression of long form PRL-R mRNA in the choroid plexus of the brain in the presence of female sex steroid and pituitary hormones for the rapid expression of maternal behaviour. Our studies also suggested that maternal behaviour can be expressed in OVX or HYPOX rats after exposure to pups for 7 days without any significant increase in brain PRL-R mRNA expression. PMID- 8708547 TI - Some effects of a low sodium intake on the expression of P450 aldosterone synthase in the hamster adrenal cortex: immunoblotting, immunofluorescent and immuno-gold electron microscopic studies. AB - In the current work we studied the effects of a low sodium intake on P450 aldosterone synthase (P450aldo) in the adrenal cortex of male hamsters by Western blotting analysis. We also investigated the zonal distribution of P450aldo with a specific antibody using immunofluorescence and immuno-gold electron microscopy. Western blotting analysis revealed a progressive induction of P450aldo in the adrenals of hamsters kept on a low sodium diet, with two-, four- and eightfold increases after 2, 4 and 21 days on the diet. Immunofluorescence microscopy showed that P450aldo was confined to the zona glomerulosa (ZG) cells. Electron microscopy showed P450aldo to be located in the mitochondria of ZG cells. When hamsters were maintained on a low sodium intake for 2, 11 and 21 days, P450aldo was still found only in the ZG; the ZG appeared either unchanged or sometimes slightly enlarged. Moreover, at days 11 and 21, the intensity of the immunofluorescent signal was much stronger in the ZG of hamsters on the low sodium intake than in controls. Hence, immunocytochemistry using the colloidal gold technique showed P450aldo to be more abundant in the mitochondria of the experimental animals than in controls. To conclude, P450aldo is present only in the ZG of hamster adrenals and sodium restriction appears to induce its expression by stimulating production within individual ZG cells rather than by stimulating a proliferation of the ZG cells. PMID- 8708548 TI - IGF-I potentiates interleukin-2 production in human peripheral T cells. AB - IGF-I stimulates the proliferation and differentiation of many cell types. In the case of T cells, IGF-I has been described to potentiate mitogen-induced DNA synthesis. We have addressed the working mechanism of IGF-I on T cell proliferation by measuring the effects of IGF-I on various stages of T cell activation. We found that IGF-I augmented the phytohaemagglutinin- and anti-CD3 induced interleukin-2 (IL2) production of human peripheral T cells before they enter the S phase of the cell cycle. Furthermore, the addition of IGF-I did not influence DNA synthesis of IL2-dependent growing T cells. PMID- 8708549 TI - Regulation of cartilage glycosaminoglycan synthesis in the rainbow trout, Oncorhynchus mykiss, by 3,3',5-tri-iodo-L-thyronine and IGF-I. AB - The actions of 3,3',5-tri-iodo-L-thyronine (T3) and recombinant human IGF-I (rhIGF-I) as well as their interaction on cartilage growth in rainbow trout (Oncorhynchus mykiss) were examined. Uptake of 3H-methyl thymidine and 35S sulfate by isolated branchial cartilage was measured as a marker for chondrocyte proliferation and sulfated glycosaminoglycan synthesis respectively. When T3 (1.0 microgram/g) was injected intraperitoneally, plasma T3 levels reached a transient peak after 1 day and decreased rapidly thereafter. Sulfate and thymidine uptake were not affected by T3 at 1 and 3 days post-injection, but at 6 days post injection both were significantly higher in T3-injected fish than those in controls. The stimulatory effects of a T3 injection on sulfate and thymidine uptake were dose-dependent over the range of 0.01, 0.1 and 1.0 micrograms/g. In vitro exposure of cartilage to T3 (0.075, 0.75, 7.5, 75 and 750 nM) for 6 days resulted in dose-dependent stimulation of sulfate uptake, with a maximum response at 7.5 nM and higher. T3 exposure (7.5 nM) for 2 or 3 days also increased sulfate uptake, but only slightly. Thymidine uptake was not clearly affected by T3. In vitro addition of rhIGF-I (0.075, 0.75 and 7.5 nM) increased sulfate uptake, but not thymidine uptake, dose-dependently. Compared with T3, rhIGF-I induced a greater maximum level of sulfate uptake: at 7.5 nM rhIGF-I increased the uptake 17-fold whereas T3 increased the uptake fourfold. When T3 (0.075, 0.75 or 7.5 nM) and rhIGF-I (0.1 or 1.0 nM) were added together, stimulative actions of T3 on sulfate uptake were largely additive to those of rhIGF-I. The results indicate that T3 as well as IGF-I are important modulators of sulfated glycosaminoglycan synthesis in rainbow trout cartilage. PMID- 8708550 TI - The focal adhesion kinase. PMID- 8708551 TI - Pioglitazone hydrochloride stimulates insulin secretion in HIT-T 15 cells by inducing Ca2+ influx. AB - Pioglitazone hydrochloride (AD-4833), one of the thiazolidinedione analogs, is a new anti-diabetic agent which improves peripheral insulin resistance in diabetic patients. We determined the direct effect of AD-4833 on insulin secretion in HIT T 15 cells. The effects of AD-4833 (10(-7) M to 10(-5) M) on insulin secretion were examined in 3 and 7 mM glucose-containing F-12 K media. The addition of 10( 5) M AD-4833 significantly increased insulin secretion in both media, but its stimulatory effect was more potent in the medium containing 7 mM glucose. The addition of 10(-5) M AD-4833 caused an immediate, significant increase in cytosolic free Ca2+ concentration ([Ca2+]i). Nifedipine at all concentrations from 10 to 1000 nM significantly attenuated insulin secretion by 10(-5) M AD 4833. In addition, 10(-5) M AD-4833 failed to stimulate insulin secretion in the CA(2+)-free Kreb's-Ringer bicarbonate buffer. These data indicated that AD-4833 stimulates in vitro insulin secretion in HIT-T 15 cells, perhaps by inducing Ca2+ influx. PMID- 8708552 TI - Beta-cell markers and autoantigen expression by a human insulinoma cell line: similarities to native beta cells. AB - In the present study we have evaluated the expression of different beta-cell markers, islet molecules and auto-antigens relevant in diabetes autoimmunity by a human insulinoma cell line (CM) in order to define its similarities with native beta cells and to discover whether it could be considered as a model for studies on immunological aspects of Type 1 diabetes. First, the positivity of the CM cell line for known markers of neuroendocrine derivation was determined by means of immunocytochemical analysis using different anti-islet monoclonal antibodies including A2B5 and 3G5 reacting with islet gangliosides, and HISL19 binding to an islet glycoprotein. Secondly, the expression and characteristics of glutamic acid decarboxylase (GAD) and of GM2-1 ganglioside, both known to be islet autoantigens in diabetes autoimmunity and expressed by human native beta cells, were investigated in the CM cell line. The pattern of ganglioside expression in comparison to that of native beta cells was also evaluated. Thirdly, the binding of diabetic sera to CM cells reacting with islet cytoplasmic antigens (ICA) was studied by immunohistochemistry. The results of this study showed that beta cell markers identified by anti-islet monoclonal antibodies A2B5, 3G5 and HISL-19 are expressed by CM cells; similarly, islet molecules such as GAD and GM2-1 ganglioside are present and possess similar characteristics to those found in native beta cells; the pattern of expression of other gangliosides by CM cells is also identical to human pancreatic islets; beta cell autoantigen(s) reacting with antibodies present in islet cell antibodies (ICA) positive diabetic sera identified by ICA binding are also detectable in this insulinoma cell line. We conclude that CM cells show close similarities to native beta cells with respect to the expression of neuro-endocrine markers, relevant beta cell autoantigens in Type 1 diabetes (GAD, GM2-1, ICA antigen), and other gangliosides. Therefore, this insulinoma cell line may be considered as an ideal model for studies aimed at investigating autoimmune phenomena occurring in Type 1 diabetes. PMID- 8708553 TI - Influence of insulin-like growth factor-binding protein-2 on plasma clearance and transfer of insulin-like growth factors-I and -II from plasma into mammary derived lymph and milk of goats. AB - Plasma clearance of insulin-like growth factors-I and -II (IGF-I and -II) and insulin-like growth factor-binding protein-2 (IGFBP-2) from lactating goats (n = 4) was determined following a single intravenous injection of the corresponding 125I-labelled human protein. Transfer of these proteins out of the vascular space was monitored by their subsequent appearance in mammary-derived lymph and milk. Clearance of 125I-IGFBP-2 from circulation was 0.37 +/- 0.06 ml/min/kg, which is markedly greater than that of 125I-IGF-I or -II (0.11 +/- 0.01 and 0.12 +/- 0.01 ml/min/kg respectively). This was also reflected in longer elimination half-lives for IGF-I (353 +/- 6 min) and -II (254 +/- 8 min) compared with IGFBP-2 (110 +/- 9 min). Three hours after injection of the 125I-labelled protein, the plasma:lymph ratio of trichloroacetic acid-precipitable radioactivity was 1.54 +/ 0.04, 3.3 +/- 0.6 and 4.1 +/- 0.4 for IGFBP-2, IGF-I and -II respectively. The form of 125I-IGFBP-2 in lymph was not different from that of plasma. Elevation of plasma concentrations of IGFBP-2 by its intravenous infusion significantly decreased plasma half-life of both IGF-I and -II (251 +/- 8 and 198 +/- 7 min respectively). Although the amount and rate of transfer of IGF into mammary derived lymph was decreased slightly by IGFBP-2, concentrations eventually obtained were not different from control. However, secretion of IGFs into milk was significantly reduced by IGFBP-2, particularly in the case of IGF-I. These results are consistent with the ability of all three compounds to cross the vascular endothelium intact and of IGFBP-2 to decrease the uptake of IGF by mammary epithelium and subsequent secretion into milk. IGFBP-2 may well have acted to target plasma IGF towards non-mammary tissues, thus explaining the more rapid plasma clearance of IGFs in the presence of elevated IGFBP-2. PMID- 8708554 TI - GH inhibition of lipogenesis and stimulation of lipolysis in sheep adipose tissue: involvement of protein serine phosphorylation and dephosphorylation and phospholipase C. AB - The intracellular signalling systems involved in the chronic insulin antagonistic, anti-lipogenic effects and also the lipolytic effect of GH have been investigated in sheep adipose tissue in an in vitro tissue culture system. During culture, chronic exposure to GH decreased the rate of lipogenesis and prevented the increase in lipogenesis induced by insulin. GH also increased glycerol release into the culture medium. GH had no acute, insulin-like effect on lipogenesis in sheep adipose tissue. Pretreatment with phorbol ester to down regulate isoforms of protein kinase C or addition of the protein serine kinase inhibitor staurosporine decreased the anti-lipogenic effect of GH while the protein serine kinase inhibitor H7 eliminated it completely. Pretreatment with phorbol ester or addition of H7 also decreased the insulin-antagonistic effect of GH on lipogenesis. Addition of the protein serine phosphatase inhibitor okadaic acid or the phosphatidyl choline phospholipase C inhibitor D609 both diminished the anti-lipogenic and insulin-antagonistic effects of GH. Chronic exposure of adipose tissue to GH had no effect on the total activity of acetyl CoA carboxylase or its activation status but it did diminish the increase in activation status induced by insulin. H7 and okadaic acid also diminished the increase in activation status of acetyl CoA carboxylase induced by insulin but did not alter the effect of GH on this variable. Okadaic acid decreased total acetyl CoA carboxylase activity. Pretreatment with phorbol ester or the addition of H7, staurosporine or okadaic acid increased glycerol release into the culture medium to the same extent as GH itself; the effects of GH and these various agents were not additive. These studies suggest that the anti-lipogenic, insulin antagonistic effects of GH involve both protein serine kinases and phosphatases, possibly including one or more isoforms of protein kinase C, and a phosphatidyl choline-specific phospholipase C. Comparison with studies by others on the GH enhancement of preadipocyte differentiation and prolactin stimulation of lipogenesis in mammary tissue suggests involvement of protein kinase C at an early stage in all three systems. In contrast, effects of okadaic acid vary with the system, suggesting the involvement of protein serine phosphatase activity in a late stage of the action of GH. The effects of GH on lipogenesis and lipolysis do not occur via identical mechanisms. PMID- 8708555 TI - Periovulatory endocrinology in high ovulating Meishan sows. AB - The present study was designed to investigate the hormone profiles (oestradiol, LH, FSH, inhibin, progesterone) in high ovulating Meishan sows (MS; n = 9) and in contemporary Large-White hybrid control sows (LW; n = 9) during the follicular phase, the periovulatory period and the early luteal phase. Ovulation rate was higher in MS than LW animals (23.7 and 16.6 respectively; P < 0.001) and overall was correlated with the area of the oestradiol peak (P < 0.05) and inhibin concentrations (P < 0.05). Both the duration of and the area of the oestradiol peak were greater in MS than LW (P < 0.01; P < 0.02), as were inhibin concentrations both before and after the LH surge (P < 0.05). Neither basal nor peak concentrations of LH or FSH differed between the breeds (P > 0.05), although FSH concentrations were correlated with the area under the oestradiol peak (P < 0.05). Finally, the time-interval from the onset of the LH surge until the rise in plasma progesterone was shorter in MS than LW (54.5 and 74.3 h respectively; P < 0.01). In conclusion, these results show for the first time that the higher ovulation rate in MS is associated with enhanced oestradiol and inhibin secretion with no significant difference in LH or FSH concentrations. The more rapid luteinization response to the LH surge by MS in terms of plasma progesterone concentrations may be important in ensuring the high level of embryo survival in this breed. PMID- 8708556 TI - Plasma clearance and tissue distribution of labelled chicken and human IGF-I and IGF-II in the chicken. AB - The metabolic clearance of chicken IGF-I (cIGF-I), cIGF-II, human IGF-I (hIGF-I), and hIGF-II was examined in the chicken using 125I-labelled growth factors. Superose-12 chromatography of plasma collected at 7.5 min post-infusion revealed peaks of radioactivity corresponding to 150 and 43 kDa and unbound tracer. Statistical analysis of trichloroacetic acid (TCA)-precipitable radioactivity in sequential plasma samples as well as following chromatography of the same samples revealed that clearance of the radiolabelled peptides followed an apparent triphasic pattern. The close similarity of the individual chromatographically defined pools in their clearance rate compared with the three components described by TCA precipitation strongly suggested their identity. Both free 125I labelled cIGF-II (3.11 min) and hIGF-II (3.01 min) were cleared at a greater rate than their IGF-I counterparts. Unbound hIGF-I was cleared at a greater rate than cIGF-I (4.45 vs 5.66 min respectively). A similar pattern for clearance was evident in the radio-labelled growth factors associated with the 43 kDa component, although at a longer half-life. There was no difference in the apparent clearance of the radiolabelled growth factors associated with the 150 kDa component between IGF-I or -II or between species. Analysis of the chromatographic profiles of radioactive IGF-I peptides complexed to serum proteins versus those bound to labelled IGF-II peptides revealed the presence of a large molecular mass binding protein in vivo. Ligand blotting of chicken serum determined that a binding protein with a mass of 70 kDa was detectable with 125I IGF-II probes only, and was not present in pig serum. In addition, tissue uptake of 125I-cIGF-I and -II was evaluated. Similar patterns of tissue distribution and uptake were observed for 125I-cIGF-I and -II, except that cIGF-II uptake by the liver exceeded that of 125I-cIGF-I at 15 min post-infusion. The rank order of tissue distribution was as follows: kidney > testis > heart > liver > pancreas > small intestine > cartilage > bursa > gizzard > leg muscle > breast muscle > brain. We conclude from these studies that the clearance of IGFs from the compartments identified in blood and the potential target tissues is dependent on their interactions with IGF-binding proteins and receptors. PMID- 8708558 TI - Molecular forms of GnRH in three model fishes: rockfish, medaka and zebrafish. AB - Three species of fish have become important in the study of reproduction and development. Rockfish are a model for developmental studies of live-bearing perch like fish, whereas medaka and zebrafish are models for developmental and genetic studies. The forms of GnRH are identified in the brains of each of these fish and in the pituitary of the rockfish to investigate the role of GnRH in reproduction. Here, we report that grass rockfish (Sebastes rastrelliger) have three forms of GnRH in brain extracts as determined by HPLC elution position and RIA. These forms are identified as sea bream GnRH, chicken GnRH-II and salmon GnRH. In contrast, only two forms of GnRH were detected in brain extracts of medaka (Oryzias latipes) and zebrafish (Brachydanio rerio): salmon GnRH and chicken GnRH II. Rockfish is distinct from medaka and zebrafish in that the most abundant form of GnRH in the rockfish pituitary is sea bream GnRH, whereas this form is absent in the other two fishes. The identification of sea bream GnRH in the rockfish brain and pituitary extracts indicates that the phylogenetic emergence of sea bream GnRH is earlier than the order Perciformes. PMID- 8708557 TI - Cytochrome P450 17 alpha-hydroxylase gene expression in differentiating rat trophoblast cells. AB - Trophoblast giant cells of the rat placenta express cytochrome P450 17 alpha hydroxylase (P450c17) and synthesize androgens. The purpose of this study was to investigate androgen production and expression of P450c17 in the Rcho-1 trophoblast cell line. These cells are capable of differentiating along the trophoblast giant cell lineage. Androstenedione production increased approximately 70-fold as Rcho-1 trophoblast cells progressed from the proliferation to the differentiation state. P450c17 enzyme activity and mRNA also showed significant increases associated with trophoblast giant cell differentiation. To study the transcriptional regulation of the P450c17 gene, the activities of a series of P450c17 promoter-luciferase reporter constructs were evaluated following transient transfection into Rcho-1 trophoblast cells. A DNA region located-98 bp upstream of the P450c17 gene transcriptional start site was the shortest promoter DNA construct consistently possessing activity in Rcho-1 trophoblast cells. Activities of longer constructs (-156 to -1560 bp) in this population of cells were significantly greater than the -98 bp promoter-reporter construct. The -476 bp P450c17 construct showed maximal promoter activity in transiently transfected Rcho-1 trophoblast cells and was developmentally activated in stably transfected Rcho-1 trophoblast cells. Activation of the cyclic AMP/protein kinase A pathway did not significantly affect P450c17 promoter activity in Rcho-1 trophoblast cells, in contrast to its effects in mouse MA-10 Leydig cells. In summary, Rcho-1 trophoblast cells are capable of endocrine differentiation and are a useful in vitro system for studying the regulation of trophoblast androgen production and P450c17 gene expression. PMID- 8708559 TI - Trophoblastic giant cells of the mouse placenta contain calbindin-D9k but not the vitamin D receptor. AB - The effects of vitamin D deficiency on the ontogeny of calcium-binding proteins (CaBPs) and the vitamin D receptor (VDR) in the placenta and yolk sac of the mouse were examined. Maternal vitamin D status did not affect the time of appearance of CaBP-D9k (9 kDa) in the yolk sac endoderm or trophoblastic giant cells (TGCs) of the placenta. VDRs were undetectable in TGCs and yolk sac endoderm, but were present in the intraplacental yolk sac. Since yolk sac endoderm and TGCs contain CaBP but not VDR, it is unlikely that CaBP synthesis and/or activity in these cells is controlled by vitamin D. The TGCs, therefore, may be involved in vitamin D-independent transplacental transfer of calcium. PMID- 8708560 TI - Nutritional regulation of gene expression of insulin-like growth factor-binding proteins and the acid-labile subunit in various tissues of rats. AB - The plasma concentration and liver mRNA content of IGF-I are regulated by the quantity and quality of dietary proteins. To determine whether the synthesis of IGF-binding proteins (BPs) is also affected by protein nutrition, we assessed plasma concentration, tissue mRNA content and liver transcription rate of each BP after rats were fed either a 12% casein or a protein-free diet for 1 week. Protein deprivation reduced the plasma concentration of IGFBP-3 and IGFBP-4 and increased that of IGFBP-1 and IGFBP-2. The mRNA content in tissues and liver transcription rates of IGFBP-3 and IGFBP-4 did not change in response to protein deprivation although their plasma concentrations decreased. The increased plasma IGFBP-1 and IGFBP-2 concentrations were explained by the increased mRNA content and transcription rate of their genes in the liver. Although IGFBP-1 mRNA was increased by protein deprivation not only in liver but also in kidney, IGFBP-2 mRNA was increased only in liver and did not increase in any other tissue examined. In addition, the liver mRNA content of the acid-labile subunit, which can form a ternary complex with IGFs and IGFBP-3, was not affected by protein deprivation. These results show that tissue-specific synthesis of each BP is regulated in a distinct way in response to protein deprivation. PMID- 8708561 TI - Expression of the steroidogenic acute regulatory protein mRNA in adrenal tumors and cultured adrenal cells. AB - The steroidogenic acute regulatory protein (StAR) has recently been shown to be a factor necessary for cholesterol transport into adrenal and gonadal mitochondria, which is the regulated, rate-limiting step in steroidogenesis. We show here that StAR mRNA is highly expressed in normal adult adrenals (n = 9), adrenocortical adenomas (n = 16), adrenal hyperplasias (n = 6), adrenocortical carcinomas (n = 6) and adrenals adjacent to tumor tissues (n = 9). There was a good correlation between the expression of StAR and the cholesterol side-chain cleavage enzyme/20,22-desmolase (P450 scc) mRNAs both in normal (r = 0.93; P < 0.01) and in tumor (r = 0.97; P < 0.001) tissues. No StAR mRNA was detected in Northern blots of liver, kidney, breast, parathyroid or phaeochromocytoma RNAs. In cultured adrenocortical cells, adrenocorticortropin (ACTH), (Bu)2cAMP, and cholera toxin increased StAR and P450 scc mRNA accumulation 6- to 18-fold, dose- and time-dependently. StAR (and P450 scc) mRNA increased relatively slowly in response to ACTH treatment, with the maximal increment at 24 h, while the mRNA of the early response gene c-fos peaked within 2 h. The protein kinase inhibitor H-7 inhibited basal and ACTH-induced StAR mRNA expression. Our results show that StAR mRNA is expressed at high levels in normal human adrenals and adrenocortical neoplasms. It is up-regulated in parallel with P450 scc by ACTH in adult adrenocortical cells, which suggests that ACTH is at least one of the key regulators of adrenal StAR expression. PMID- 8708562 TI - The ovine pancreatic protein which binds insulin-like growth factor binding protein-3 is procarboxypeptidase A. AB - Insulin-like growth factor binding protein-3 (IGFBP-3) is known to modulate the actions of insulin-like growth factors (IGF)-I and -II at the level of the cell. Proposed mechanisms include association of IGFBP-3 with cell surface proteoglycan, with cell surface binding proteins, proteolysis and/or internalization of IGFBP-3. In previous studies we have characterized a protein of 40 kDa in extracts of ovine pancreas and muscle which binds IGFBP-3 on ligand blot analyses. This paper reports the identity of the pancreatic species as procarboxypeptidase A (peptidyl-L-amino acid hydrolase, E.C. 3.4.17.1; proCPA). Identity was established by amino terminal sequence analysis, binding studies with pure bovine carboxypeptidase A (CPA) and observations that the binding activity was present in pancreatic secretions consistent with the role of proCPA as a secretory zymogen. The binding activity was inhibited by unlabelled IGFBP-3 at high doses (10 micrograms/ml) and reduced but not abolished by preincubation of 125I-IGFBP-3 with excess IGF-I. Digestion of 125I-IGFBP-3 with mature CPA produced a 26 kDa product. Modification of IGFBP-3 by CPA or binding to proCPA may provide a mechanism for modulation of IGFBP activity and hence IGF action. PMID- 8708563 TI - In vivo manipulation (depletion versus activation) of testicular macrophages: central and local effects. AB - Testicular macrophages are a relevant cell type for the regulation of Leydig cell steroidogenesis. The availability of liposome technology allows in vivo manipulation of macrophages in order to analyze their role in the regulation of the hypothalamic-pituitary-testicular axis. In this study, adult (70 days of age) and prepubertal (22 days of age) rats were injected intratesticularly with liposomes containing either dichloromethylene diphosphonate (C12MDP) to deplete testicular macrophages or muramyl tripeptide (MTP-PE) to activate them. Control rats were injected with the corresponding volumes of 0.9% NaCl. Animals were killed 10 days after treatment. Adult rats injected bilaterally or unilaterally with C12MDP liposomes showed increased serum LH and testosterone concentrations, as well as increased testosterone concentrations in the testicular interstitial fluid. In unilaterally injected rats, testosterone concentrations in the interstitial fluid were higher in the macrophage-containing testes than in the contralateral, macrophage-depleted testes. Adult rats treated bilaterally with MTP-PE liposomes showed increased numbers of testicular macrophages, whereas the number of Leydig cells was unchanged. Serum LH concentrations were decreased, but no changes were found in testosterone concentrations. Prepubertal rats treated bilaterally with C12MDP liposomes showed decreased numbers of Leydig cells. However, serum LH and testosterone concentrations were increased. Otherwise, prepubertal rats treated bilaterally with MTP-PE liposomes showed increased numbers of macrophages and Leydig cells, as well as increased serum testosterone concentrations. These data suggest that testicular macrophage-derived factors act at two different levels in the pituitary-testicular axis: first, at a central level by inhibiting LH secretion, and secondly, at a local level by stimulating Leydig cell steroidogenesis. PMID- 8708564 TI - IGF-I and IGF-binding protein-3 in plasma of GH-deficient rats. AB - The majority of IGF-I circulates in a large (150 kDa) ternary complex with IGF binding protein-3 (IGFBP-3) and a non-IGF-binding acid-labile subunit. The secretion of ternary complex into the circulation from liver has been considered to be GH-dependent; however, recent data indicate that GH does not directly regulate hepatic IGFBP-3 synthesis. To examine the role of insulin in regulating plasma IGFBP-3 levels, postpubertal male GH-deficient (dw/dw) rats were treated every 8 h with injections (s.c.) of 0.9% saline, 20 micrograms insulin/day, 200 micrograms hIGF-I/day, or 20 micrograms insulin/day plus 200 micrograms hIGF I/day, for 10 days with the animals being killed 2-3 h after the final injection. Hypoglycaemia was not observed in any of the treatment groups. hIGF-I treatment increased longitudinal growth and weight gain (P < 0.05), while insulin treatment had no effect. Plasma IGF-I levels were increased in groups treated with hIGF-I (P < 0.05), while insulin treatment resulted in a reduction (P < 0.05): saline = 267.1 +/- 15.6 (ng/ml +/- S.E.M.), insulin = 219.3 +/- 17.5, hIGF-I = 391.7 +/- 17.6, insulin plus hIGF-I = 357.5 +/- 31.8. Hepatic IGF-I mRNA expression was increased in insulin-treated dw/dw rats in comparison with hIGF-I-treated animals (P < 0.05) but not in comparison with saline control or the combined treatment groups. Plasma levels of intact IGFBP-3, measured by ligand blot analysis, were increased in all treatment groups compared with saline (P < 0.05): saline = 100.0 +/- 9.4% (% of saline +/- S.E.M.), insulin = 149.9 +/- 17.5%, hIGF-I = 191.4 +/- 17.3%, insulin plus hIGF-I = 205.4 +/- 15.3%. The levels of the 28/32 kDa IGFBPs and IGFBP-4 in plasma were increased by hIGF-I treatment (P < 0.05) but not by insulin treatment. Hepatic specific 125I-bovine GH binding was not significantly different in any of the treatment groups. This study provides the first evidence in nondiabetic animals that insulin regulates hepatic IGF-I mRNA expression, plasma IGF-I and plasma IGFBP-3 levels in the GH-deficient state without changes in hepatic GH receptors. The divergent response of plasma IGF-I and IGFBP-3 levels to insulin treatment in the present study may indicate an effect of insulin on the clearance of IGF-I from the circulation. PMID- 8708565 TI - Superior potency of infused IGF-I analogues which bind poorly to IGF-binding proteins is maintained when administered by injection. AB - The relative potency of IGF-I and the analogue LR3IGF-I to either promote growth or reverse catabolism in rats when administered by injection rather than by continuous infusion has been examined. LR3IGF-I has very low affinity for the IGF binding proteins in the rat and hence is cleared from the circulation more quickly than is IGF-I. Experiments were performed in normal growing rats (150 g body weight) and in rats made catabolic by dexamethasone infusion (20 micrograms/day). IGFs or vehicle were delivered subcutaneously for 7 days either by continuous infusion via osmotic pumps or by injection once or twice daily at 320 and 400 micrograms/day in normal and catabolic rats respectively. As expected, continuous infusion of IGFs showed greater efficacy than either of the injection modes especially in its anti-catabolic actions. When infused continuously LR3IGF-I was generally 1.5- to 2-fold more potent than IGF-I for changes in body weight gain, visceral organ weights and feed use efficiency. Notably, LR3IGF-I remained more potent than IGF-I in several of these effects even when the peptides were given by once-daily injection. In addition, N tau methylhistidine excretion by dexamethasone-treated rats was reduced to a threefold greater extent by injected LR3IGF-I than by injected IGF-I. Notwithstanding these effects, LR3IGF-I was barely equipotent with IGF-I for reversal of carcass muscle loss in dexamethasone-treated rats. Despite its more rapid clearance from the circulation, injected LR3IGF-I retains superior potency to injected IGF-I for several actions, albeit the potency is much reduced compared with continuous infusion. Thus our data indicate that use of IGF analogues which have low affinity for binding proteins may have advantages in potency and/or tissue specificity where IGFs are necessarily administered by injection. PMID- 8708566 TI - Recombinant FSH-induced follicle development in immature rats treated with an LHRH antagonist:a direct effect of RU486 on follicular atresia. AB - To investigate whether the progesterone antagonist RU486 has a direct effect on ovarian function, it was administered to immature female rats rendered hypogonadotrophic by administration of an LHRH antagonist and in which follicle development was stimulated by recombinant human FSH (recFSH). In the first experiments the effects of LHRH antagonist and recFSH on follicle growth were evaluated. Female rats of 22 days of age were injected with an LHRH antagonist (Org 30276; 500 micrograms/100 g body weight) every other day. This treatment resulted in a tenfold decrease in serum LH concentrations and a twofold decrease in serum FSH concentrations at day 30 and caused a reduction in the number and size of antral follicles. Treatment with recFSH (Org 32489) twice daily from day 26 for 4 days in a total dose ranging from 5 to 20 IU/animal increased the number and size of antral follicles in a dose-related manner and resulted after 20 IU recFSH in a tenfold increase in the concentration of inhibin in serum and ovaries at day 30. Once it was established that LHRH antagonist treatment in immature rats could be used to study the effects of gonadotrophins or steroids on follicle function, this animal model was used to study the effects of RU486 on the ovary. RU486 was administered (twice daily for 4 days, 1 mg/injection) to LHRH antagonist-treated rats in which follicular growth and differentiation were stimulated by 10 IU recFSH or by 10 IU recFSH plus 0.5 IU human chorionic gonadotrophin (hCG). RU486 had no effect on circulating levels of LH and FSH, but stimulated follicular atresia both in rats treated with recFSH alone and in rats treated with recFSH and hCG. Inhibin concentrations both in serum and ovaries were significantly increased after hCG treatment. RU486, however, did not increase inhibin in the rats treated with recFSH and in those treated with recFSH and hCG. In summary, the present study has demonstrated that (1) immature rats treated with an LHRH antagonist can be used to study the effects of gonadotrophins and steroids on follicular function and (2) RU486 has a direct stimulatory effect on follicular atresia. PMID- 8708567 TI - Monoclonal antibodies specific for the binding site on the LH receptor alter progesterone production in cultured granulosa cells. AB - Monoclonal antibodies (mAbs) specific for the LH receptor (LHR) were generated through a modified auto-anti-idiotypic approach in which human chorionic gonadotropin (hCG) was used as the immunogen followed by cyclophosphamide to induce anti-idiotypic antibodies. The purpose of this study was to investigate the effectiveness of these antibodies to alter progesterone production in porcine granulosa cells in vitro. Anti-LHR mAbs were incubated with granulosa cells in the presence or absence of a stimulatory dose of hCG. Progesterone output by treated cells was measured using a RIA procedure. Most of the mAb could inhibit stimulated progesterone production by cultured granulosa cells. It was speculated that two possible mechanisms may cause the inhibition effect observed. Several of the antibodies appeared to block hCG binding thus removing the stimulatory effects of hCG. However, the most potent inhibiting mAbs for progesterone production had little or no effect on hCG binding, suggesting that some other mechanism was responsible for the observed inhibition. In addition, several of the antibodies were found to have a stimulatory effect on progesterone production by granulosa cells even in the absence of a stimulating dose of hCG. It is proposed that these antibodies were able to mimic hCG. PMID- 8708568 TI - A study of the luteolytic mechanism of the antiprogesterone RU486 during the late luteal phase in pseudopregnant rats. AB - The purpose of this study was to examine the possible mechanism through which RU486 induces luteolysis during the late-luteal phase in pseudopregnant (PSP) rats. PSP rats received a subcutaneous injection of RU486 in sesame oil (5 mg/kg body weight) or sesame oil alone once a day between day 9 and day 11 of pseudopregnancy. Serial blood samples were collected on days 5, 9, 10, 11 and 12 and assayed for progesterone content. To examine the possible action of RU486 through a uterine and/or a pituitary (prolactin-dependent) mechanism, PSP rats and chronic hysterectomized PSP rats which had been hysterectomized before PSP induction received a subcutaneous injection of RU486 in sesame oil (5 mg/kg body weight), sesame oil alone, prolactin in 50% polyvinylpyrrolidone (15 IU/day), or RU486 and prolactin once a day between day 9 and day 11 of pseudopregnancy. Serial blood samples were collected on days 5, 9, 10 and 11 and assayed for progesterone content. Blood samples were also collected at 0400 h on day 12 and used for prolactin and progesterone determinations. To examine the direct effect of RU486 on corpus luteum and/or pituitary, hysterectomized rats underwent hypophysectomy and pituitary autotransplantation on dioestrus 1 and received a subcutaneous injection of RU486 in sesame oil or sesame oil alone for 3 days between day 21 and day 23 after surgery. Serial blood samples were collected on days 10, 21, 22, 23 and 24 and assayed for progesterone and prolactin contents. In ordinary PSP rats, serum progesterone levels were significantly (P < 0.01) lower in the RU486-treated group than in the control group (9 +/- 1 vs 53 +/- 7 ng/ml; mean +/- S.E.M.) on day 11. Serum prolactin levels at 0400 h on day 12 of pseudopregnancy were significantly (P < 0.05) lower in the RU486-treated group than in the control group (16 +/- 4 vs 154 +/- 44 ng/ml; mean +/- S.E.M.). The concomitant prolactin treatment reversed the luteolytic effects of RU486 on day 11 of pseudopregnancy. In hysterectomized PSP rats, RU486 also suppressed serum prolactin levels, and the concomitant prolactin treatment again reversed the luteolytic effects of RU486. In hysterectomized rats which were hypophysectomized and pituitary autotransplanted, RU486 treatment did not induce any significant changes in serum progesterone and prolactin levels. These results indicated that RU486 induced luteolysis during the late-luteal phase in PSP rats by suppressing prolactin secretion via a hypothalamic mechanism. PMID- 8708570 TI - A tribute to Charles Richard Taylor. Charles P. Lyman professor of biology at Harvard University. 8 September 1939 to 10 September 1995. PMID- 8708569 TI - Inhibitory effects of fumagillin and its analogue TNP-470 on the function, morphology and angiogenesis of an oestrogen-induced prolactinoma in Fischer 344 rats. AB - The process of angiogenesis occurs in many physiological states, but it is also essential for the growth of solid tumours and metastasis formation. An abnormal arterial vascularization has been shown in prolactin-secreting pituitary adenomas induced by prolonged treatment with oestrogens in Fischer 344 (F344) rats. It is thought that anti-angiogenic agents might be useful in therapy for these tumours. Fumagillin and its analogue TNP-470 are known to inhibit endothelial cell proliferation selectively, but their effect on lactotroph cell secretory function and prolactinoma formation has not yet been described. The aim of the present study was to examine the effects of fumagillin and TNP-470 on prolactin secretion, and morphological and vascular changes within the anterior pituitary in long-term oestrogen-treated male F344 rats in vivo and in vitro. As expected, 7 weeks after s.c. implantation of Silastic tubes containing 10 mg diethyl stilboestrol (DES), a very high rise in serum prolactin levels was found. Both angiogenesis inhibitors injected s.c. at doses of 10 mg/kg body weight for 24 days attenuated the stimulatory effect of DES on prolactin production and release. They also diminished prolactin cell density and inhibited cell proliferation expressed as the number of anterior pituitary cells labelled with bromodeoxyuridine (BrdU), but the effect of TNP-470 was minor compared with fumagillin. Both angioinhibitors suppressed neo-vascularization within the anterior pituitary with similar potency but, on the other hand, they did not affect DES-induced increases in prolactin secretion from cultured rat pituitary cells and cell proliferation in vitro. In conclusion, our results provide strong evidence for the anti-tumour and anti-prolactin activity of angiogenesis inhibitors in the experimentally oestrogen-induced pituitary adenoma; this might be mediated indirectly through the inhibition of angiogenesis. PMID- 8708571 TI - Design of the oxygen and substrate pathways. I. Model and strategy to test symmorphosis in a network structure. AB - This first paper in a series develops a model of structure-function relationships for the oxygen and substrate pathways of oxidative metabolism in working muscle. This will be used in the subsequent experimental papers in asking how biological structures are designed if they serve more than one function and whether one function can be served by more than one structural pathway. We have used the concept of symmorphosis to address this question; in its original form, it postulates that no more structure is built and maintained at each step in a pathway than is required to meet functional demands. The concept of symmorphosis was developed to deal with the problem of modelling the pathway for oxygen from the environment to mitochondria, essentially a single series of interconnected transfer steps. In the present context, the application of this concept is more complex. Both oxygen and substrates are transported directly from the blood to the mitochondria in what appear to be shared steps. The flows along this direct pathway are adjusted during muscular work. However, substrates have an additional option. They can be stored intracellularly as lipid droplets or glycogen, and thus their supply to mitochondria can occur in two steps separated in time: from capillaries to stores during rest, and from stores to mitochondria during work. The integrated pathways have a network structure and the functional flows are partitioned to different branches of the network, and we must ask whether the partitioning of fluxes is related to design constraints. The principle of symmorphosis predicts that the best use is made of the available options and that the design of each step is matched to the specific functional demand in view of a balance to be achieved over the entire network. This will be tested in subsequent papers by determining maximal flows for oxygen, carbohydrates and lipids through each of the transport steps and their respective structural capacities, comparing dogs and goats, animals of the same size whose maximal oxidative capacities differ by more than twofold. Finally, we will ask whether the principle of symmorphosis can be extended to apply to network systems. PMID- 8708572 TI - Design of the oxygen and substrate pathways. II. Defining the upper limits of carbohydrate and fat oxidation. AB - This paper quantifies maximal flows of carbohydrates and lipids through the pathways supplying the mitochondria. Maximal flow rates are the main functional parameter used in testing the principle of symmorphosis, which states that structural capacities are quantitatively matched to functional demand. Only under rate-limiting conditions will all of the structural capacity be used. Dogs and goats were compared to obtain large differences in absolute rates. We exercised the animals for long enough to reach steady-state O2 and CO2 exchange rates at intensities eliciting 40%, 60% and 85% of the maximal rate of oxygen consumption (MO2max). We then calculated rates of fat and carbohydrate oxidation from the ratio of CO2 produced to O2 consumed (the respiratory exchange ratio). The dog's Mo2max was more than twice that of the goat (6517 versus 3026 mumol O2 kg-1 min 1). We found the same pattern of fuel selection as a function of exercise intensity in both species, and it appears to be general to mammals. Maximal rates of fat oxidation were reached at 40% exercise intensity, where 77% of the energy was supplied by fat. As exercise intensity increased, all additional energy was supplied by carbohydrates. We conclude that the partitioning of fuel supply to the fat and carbohydrate pathways follows the same pattern in both dogs and goats. PMID- 8708573 TI - Design of the oxygen and substrate pathways. III. Partitioning energy provision from carbohydrates. AB - This paper quantifies maximal fluxes through the pathway supplying carbohydrates to the mitochondria of muscle cells. Continuous infusions of D-[3-(3)H]glucose together with indirect calorimetry were used to investigate the partitioning of the supply of carbohydrates through the two branches of the pathway: from circulating glucose and from glycogen stores within the muscle cells to the mitochondria. The relative contribution of circulating glucose to total carbohydrate oxidation was small, accounting for only 13% and 23% of the carbohydrate oxidized at exercise intensities approaching MO2max in dogs and goats, respectively. Unexpectedly, maximal rates of circulating glucose oxidation were nearly the same in the two species (when expressed in absolute terms; dog:goat ratio = 1.2), despite the 2.2-fold difference in aerobic capacity. We conclude that the glycogen stores in the muscle cells are the major source of substrates at maximal rates of oxidation, supplying 60-70% of the total energy. Furthermore, it is this branch of the carbohydrate pathway that is adapted to the large difference in aerobic capacity between dogs and goats. PMID- 8708574 TI - Design of the oxygen and substrate pathways. IV. Partitioning energy provision from fatty acids. AB - This paper quantifies the fluxes of fatty acids through the pathways supplying muscle mitochondria with oxidative fuel in exercising dogs and goats. We used continuous infusions of 1-[14C]palmitate and indirect calorimetry to measure fatty acid supply from two sources: the circulation and the triglyceride stores within the muscle cells. Our goal was to determine maximal flux through these two branches of the lipid pathway as key functional parameters for testing the principle of symmorphosis, i.e. that structural capacity is quantitatively matched to functional demand in the oxidative substrate pathways. It is under these rate-limiting conditions that we predict that all of the structural capacity will be used. Maximal rates of fatty acid oxidation were reached at low exercise intensities of 40% Mo2max. Fatty acids from the circulation supplied only a small fraction (15-25%) of the total fat oxidized under these conditions. Although dogs were able to oxidize circulatory fatty acids faster than goats, maximal rates were not in proportion to the 2.2-fold difference in aerobic capacity between the two species. Dogs compensated for their relatively lower use of circulatory fatty acids by oxidizing more triglycerides from lipid droplets in their muscle cells. We conclude that fatty acids from intramuscular triglyceride stores are a major source of fuel during maximal rates of lipid oxidation. Furthermore, it is this branch of the fatty acid pathway that is adapted to the large difference in aerobic capacity between dogs and goats. PMID- 8708575 TI - Design of the oxygen and substrate pathways. V. Structural basis of vascular substrate supply to muscle cells. AB - This paper quantifies the structural capacity of the transport steps for oxygen, glucose and fatty acids from the blood in capillaries to the cytosol of muscle cells and compares it with maximal rates of oxygen and substrate transport measured in the same animals and reported in the preceding papers of this series. Dogs have relatively more muscle per unit body mass than goats (37 versus 26%), but the maximal rate of oxidation per gram of muscle is still larger in the dog by a factor of 1.55. The maximal rates of substrate supply from the circulation are similar in both species. We predict that these differences in physiological parameters should be matched by proportional differences in structural capacity. We find that capillary volume and surface area are matched to maximal oxygen demand. The rate of vascular substrate supply is proportional neither to the capillary surface area nor to the length of intercellular junctions. The sarcolemmal surface area per gram of muscle is the same in both species. Using the physiological data presented in the companion papers of this series, we have calculated the maximal flux densities of circulatory glucose and fatty acids across the capillary wall and the sarcolemma. We find, for both substrates, that the flux densities across the sarcolemma reach a maximum at nearly the same level and at low exercise intensities in both species. In contrast, the flux densities across the capillary surface and the endothelial junctions are higher in goats than in dogs. We conclude that the capillaries are designed for O2 supply up to maximal rates of oxidation but not for the supply of the substrates (glucose and fatty acids) at the rates required at high exercise intensities. These are limited by the transport capacities of the sarcolemma. PMID- 8708576 TI - Design of the oxygen and substrate pathways. VI. structural basis of intracellular substrate supply to mitochondria in muscle cells. AB - This paper quantifies the structures involved in the transport and oxidation of carbohydrates and fatty acids within the muscle cell. The structural capacity is measured on whole-body random samples of the musculature of dogs and pygmy goats and compared with maximal rates of oxygen consumption and substrate oxidation. Comparing dogs and goats of the same body size provided a 1.55-fold difference in the maximal rate of oxidation when related to muscle mass. As in previous studies, we found that the volume of mitochondria was approximately proportional to aerobic capacity. The maximal glucose flux from intracellular stores to mitochondria is 1.6 times greater in the dog than in the goat; we find that the amount of glycogen stored in the muscle cells is 4.2 times as great in the dog, but part of the intracellular glycogen pool is used for anaerobic rather than for oxidative metabolism. The maximal fatty acid flux from intracellular stores to mitochondria is 1.5 times larger in the dog, and the amount of lipid stored is 2.3 times as great in the dog. Every lipid droplet is in direct contact with the outer membrane of a mitochondrion and the contact surface area is 3.6 times greater in the dog than in the goat. Additional measurements are needed to investigate the role of structural limitation at this step. The amount of substrates stored intracellularly in the muscle cells of the dog is about twice as much as would match the differences in the maximal rates of utilization. This allows the endurance-specialized dogs to run for longer periods at higher rates of oxidation. PMID- 8708577 TI - Design of the oxygen and substrate pathways. VII. Different structural limits for oxygen and substrate supply to muscle mitochondria. AB - This paper integrates the results of a series of studies on the supply of O2 and substrates for oxidative muscle metabolism and draws conclusions on the role of structural design in partitioning and limiting substrate supply. The studies compared dogs and goats exercising at different intensities and combined physiological, biochemical and morphometric investigations. In both species, the rate of fatty acid oxidation reached an upper limit at low exercise intensities, and only glucose consumption was increased at higher exercise intensities. The supply of both glucose and fatty acids from the capillaries reached maximal rates at low exercise intensities; this limitation is related to the design of the sarcolemma as calculations suggest that the endothelium introduces only a small resistance to substrate flux. From these findings, it appears that the capillaries are designed to satisfy O2 supply up to maximal O2 demand. The increase in substrate supply to the mitochondria at higher exercise intensities is achieved by drawing on intracellular stores of glycogen and lipids. The size of these stores is larger in dogs than in goats, providing the athletic species with twice the fuel reserves. These findings are interpreted on the basis of a network model with fluxes partitioned between direct and indirect pathways and with some structures shared by more than one function. Whereas O2 is supplied through a direct pathway, the supply of both substrates is split temporally to allow, during exercise, immediate fuel supply to the mitochondria from intracellular stores; these are replaced from the vasculature, during periods of rest, to a size commensurate with high rates of combustion. Considering this complexity, we conclude that the results are compatible with the principle of symmorphosis applied to a network structure and that the adjustment of design to functional demand involves different structures for O2 and for substrates. PMID- 8708578 TI - Optomotor control of course and altitude in Drosophila melanogaster is correlated with distinct activities of at least three pairs of flight steering muscles. AB - Flight control in the fruitfly Drosophila melanogaster is achieved by minute sets of muscles on either side of the thorax. Control responses of wings and muscles were elicited during fixed flight by moving a striped pattern in front of the eyes. For example, pattern motion from the lower right to the upper left signals to the test fly a rotatory course deviation to the right and simultaneously a translatory altitude displacement downwards. The counteracting response to the displacement of the retinal image is an increase in thrust and lift on the right, accomplished mainly by increasing the wingbeat amplitude (WBA) on that side. A comparison of such responses with the simultaneously recorded action potentials in the prominent basalar muscles M.b1 and M.b2 and axillary muscles M.I1 and M.III1 on either side suggests that three of these muscles act on the WBA more or less independently and contribute to the optomotor control of course and altitude. During flight, M.b1 is almost continuously active with a frequency equal to or slightly below 1 spike per wingbeat cycle. The spikes occur within a narrow phase interval of this cycle, normally at the beginning of the transition from upstroke to downstroke. However, the visual stimulus described above causes a substantial phase lead in M.b1 on the right; the spikes occur shortly before the end of the upstroke. Such phase shifts are accompanied by comparatively smooth 'tonic' responses of the WBA. The activities of M.b2 and M.I1 are normally very low. However, the stimulus described above activates M.b2 on the right in a phase interval approximately two-thirds into the upstroke and M.I1 on the left in a phase interval at the beginning of the downstroke. The spikes tend to occur in bursts. These bursts are correlated with WBA-increasing 'hitches' (rapid changes in amplitude) on the right and WBA-decreasing hitches on the left. As fast 'phasic' responses, the burst-induced hitches are likely to account for the course-controlling 'body saccades' observed during free flight. For unknown reasons, M.I1 is activated by pattern motion but cannot conceivably assist the other muscles in altitude control. Unlike its homologues in larger flies (Musca domestica, Calliphora erythrocephala), M.III1 does not participate in optomotor flight control. Its activation seems to support the termination of flight and wing retraction at rest. The essential properties of the three pairs of muscles M.b1, M.b2 and M.I1 resemble those found in larger flies; the muscles are controlled by motion detectors with muscle-specific 'preferred directions' in the hexagonal array of retinal elements. Optomotor control of the three pairs of muscles in Drosophila melanogaster could explain most, but not all, of the WBA responses recorded so far. PMID- 8708579 TI - The initiation of pre-ecdysis and ecdysis behaviors in larval Manduca sexta: the roles of the brain, terminal ganglion and eclosion hormone. AB - Each larval molt of Manduca sexta culminates in the sequential performance of pre ecdysis (cuticle loosening) and ecdysis (cuticle shedding) behaviors. Both behaviors are thought to be triggered by the release of a peptide, eclosion hormone (EH), from brain neurons whose axons extend the length of the nervous system. EH bioactivity appears in the hemolymph at the onset of pre-ecdysis behavior, and EH injection can trigger pre-ecdysis and ecdysis behaviors prematurely. The present study examined the effects of removing or disconnecting portions of the central nervous system prior to the time of EH release on the initiation of pre-ecdysis and ecdysis behaviors at the final larval molt. We found that the initiation of pre-ecdysis abdominal compressions at the appropriate time required the terminal abdominal ganglion (AT) but not the brain; the initiation of pre-ecdysis proleg retractions at the appropriate time required neither the AT nor the brain; the initiation of ecdysis at the appropriate time usually required the brain but did not require the AT; and premature pre-ecdysis (but not ecdysis) could be elicited in isolated abdomens by injection of EH. Finally, pre-ecdysis behavior performed by brainless larvae was not associated with the normal elevation of EH bioactivity in the hemolymph or the normal loss of EH immunoreactivity from peripheral neurohemal release sites. PMID- 8708580 TI - Cardiorespiratory synchrony in turtles. AB - Many reptiles, particularly diving species, display characteristic cardiovascular changes associated with lung ventilation (cardiorespiratory synchrony). Previous studies on freshwater turtles show that heart rate and pulmonary blood flow rate (Qpul) increase two- to fourfold during ventilation compared with breath-holding, and some studies report concomitant decreases in systemic blood flow rate (Qsys). The primary aim of this study was to provide a detailed description of cardiorespiratory synchrony in free-diving and fully recovered turtles (Trachemys scripta). During breath-holds lasting longer than 5 min, Qpul averaged 15 ml min 1 kg-1 and increased more than threefold to a maximum value of 50 ml min-1 kg-1 during ventilation. Qsys also increased during ventilation compared with during breath-holds lasting longer than 5 min (from 44 to 73 ml min-1 kg-1 during ventilation). Neither Qpul nor Qsys was affected by the number of breaths in the ventilatory periods. Changes in Qpul and Qsys were accomplished entirely through a significant increase in heart rate during ventilation, while total stroke volume (systemic+pulmonary) remained constant. Irrespective of the ventilatory state, Qsys exceeded Qpul by 20-30 ml min-1 kg-1. Nevertheless, because Qpul increased relatively more than Qsys during ventilation, Qpul/Qsys increased from 0.29 during apnoea to 0.80 during lung ventilation. This study confirms cardiorespiratory synchrony in the turtle Trachemys scripta but, in contrast to earlier studies, a net right-to-left cardiac shunt prevailed regardless of ventilatory state. PMID- 8708581 TI - Do Australian desert frogs co-accumulate counteracting solutes with urea during aestivation? AB - Australian desert frogs of the genera Neobatrachus, Cyclorana and Heleioporus experience significant dehydration, and iono- and osmoconcentration, during aestivation in the laboratory and accumulate substantial amounts of urea (100-200 mmol)(l-1). We expected a priori that aestivating frogs probably would not need to accumulate balancing osmolytes but would accumulate trimethylamine oxide (TMAO) or betaine as counteracting solutes to urea. These aestivating frogs did not co-accumulate a substantial quantity of any particular balancing osmolyte or counteracting solute, such as a methylamine [TMAO, trimethylamine amine (TMA), betaine, sarcosine, glycerophosphorylcholine (GPC)] or polyol (inositol, mannitol, sorbitol) in plasma or muscle relative to urea accumulation. However, for aestivating frogs, the total concentration of all measured methylamines and polyols (TMAO + TMA + betaine + sarcosine + GPC + inositol) in muscle was approximately 35-45 mmol kg-1, and so it is possible that all of these solutes have a combined counteracting osmolyte role in aestivating frogs at a ratio to urea of approximately 1:2.5, as has been described for elasmobranch fishes. Alternatively, the absence of substantial co-accumulation with urea of any particular solute suggests that aestivating frogs might not require any major extracellular or intracellular counteracting solutes (TMAO, betaine, GPC). The enzyme systems of these aestivating frogs may be insensitive to the perturbing effects of urea, or the perturbing effects of accumulated urea may be a mechanism for metabolic depression, during aestivation. PMID- 8708582 TI - The modulus of elasticity of equine hoof wall: implications for the mechanical function of the hoof. AB - During normal weight-bearing and locomotion, the equine hoof wall deforms in a consistent pattern; the proximal dorsal wall rotates caudo-ventrally about the distal dorsal border and there is latero-medial flaring posteriorly. The aim of this study is to examine whether there are regional differences in the modulus of elasticity of hoof wall material and whether such differences correlate with the pattern of deformation which occurs in vivo. The modulus of elasticity of equine hoof wall was determined in tension and compression for samples from six forefeet. Samples were tested at the mid-point of the inner and outer halves of the wall thickness at two positions along the proximo-distal axis of the dorsal wall, and from the mid-point of its thickness at the lateral and medial quarters. Test samples were oriented both parallel and perpendicular to the tubules that characterise the microstructure of the wall. The colour of each sample was noted, and the moisture content measured. The range in the mean modulus of elasticity for all samples and tests was 460-1049 MPa, the dorsal outer wall having the highest values, the dorsal inner wall the lowest, and the quarters having intermediate values. The mean value obtained for the quarters was similar to the average of the values for the dorsal inner and outer walls. At all sites, the modulus of elasticity was marginally higher in compression than in tension, possibly owing to microstructural defects. The difference in stiffness between the outer wall and the inner wall was inversely related to moisture content. The difference in stiffness between the dorsal outer and inner walls demonstrates that the equine hoof wall has a comparatively rigid external capsule with a lining of lower stiffness. This arrangement presumably provides some stress protection to the internally adjacent living tissues. The similarity in stiffness between the samples from the quarters and the mean of the two dorsal wall sites suggests that the wall at the quarters has a similar change in stiffness across its thickness as the dorsal wall. However, the reduced thickness of the wall at the quarters compared with the dorsal wall means that, functionally, the quarters are more flexible than the dorsal wall. This will facilitate the flaring of the lateral and medial walls which occurs during weight-bearing. Anisotropy was evident only in tensile tests of the dorsal wall samples. Contrary to popular assertions that white hooves are mechanically inferior, horn pigmentation had no detectable effect on stiffness. PMID- 8708583 TI - Effect of engineering Hsp70 copy number on Hsp70 expression and tolerance of ecologically relevant heat shock in larvae and pupae of Drosophila melanogaster. AB - To determine how the accumulation of the major Drosophila melanogaster heat-shock protein, Hsp70, affects inducible thermotolerance in larvae and pupae, we have compared two sister strains generated by site-specific homologus recombination. One strain carried 12 extra copies of the Hsp70 gene at a single insertion site (extra-copy strain) and the other carried remnants of the transgene construct but lacked the extra copies of Hsp70 (excision strain). Hsp70 levels in whole-body lysates of larvae and pupae were measured by ELISA with an Hsp70-specific antibody. In both extra-copy and excision strains, Hsp70 was undetectable prior to heat shock. Hsp70 concentrations were higher in the extra-copy strain than in the excision strain at most time points during and after heat shock. Pretreatment (i.e. exposure to 36 degrees C before heat shock) significantly improved thermotolerance, and this improvement was greater and more rapid in larvae and pupae of the extra-copy strain than in those of the excision strain. The experimental conditions resemble thermal regimes actually experienced by Drosophila in the field. Thus, these findings represent the best evidence to date that the amount of a heat-shock protein affects the fitness of a complex animal in the wild. PMID- 8708585 TI - Hepatitis C virus transmission in hospitals. PMID- 8708584 TI - Synergistic action of neuropeptide Y and adrenaline in the eel atrium. AB - To investigate the influence of neuropeptide Y (NPY) on heart function in a relatively simple model system, the effects of eel NPY (eNPY) on the eel atrium were examined. Eel NPY enhanced the contractile force of the isolated atrium in a dose-dependent manner, without altering the rate of contraction. Although adrenaline also exerts a positive inotropic effect, the effect of eNPY was not blocked by the beta 1-adrenoceptor antagonist betaxolol, indicating that eNPY does not act via adrenaline release. When eNPY and adrenaline were applied simultaneously, their effects were additive at lower concentrations but not at higher concentrations. The plateau reached at high concentrations suggests that these two regulators act through a common signal transduction process. One candidate for this is an elevation of the concentration of intracellular free Ca2+ ([Ca2+]i), since treatment with eNPY or adrenaline enhanced [Ca2+]i, as assessed by fluorescence of Calcium Green-1. The increase in [Ca2+]i after eNPY and adrenaline treatment is presumably due to Ca2+ influx from the external medium, since the effect was greatly reduced in Ca(2+)-free Ringer's solution and after treatment with verapamil, a Ca2+ channel blocker. Although both eNPY and adrenaline enhanced the atrial contractile force, the time courses were different, with the effect of eNPY being gradual and long-lasting, and that of adrenaline being immediate and transient. It is plausible, therefore, that eNPY and catecholamine(s) act synergistically to cause a long-lasting enhancement of contraction force if the two regulators arrive at the atrium simultaneously. The present study also demonstrates that the eel heart contains eNPY. PMID- 8708586 TI - Changes in cardiovascular risk factors by combined pharmacological and nonpharmacological strategies: the main results of the CELL Study. AB - OBJECTIVE: The objectives of the Cost Effectiveness of Lipid Lowering (CELL) study were twofold: (i) to evaluate the effect on overall cardiovascular risk of two types of health care advice ("usual' and "intensive') given in primary care, with or without pharmacological medication, with the target being to attain a moderate decrease in cholesterol; (ii) to evaluate the ritual of daily medication on compliance with the health care advice. DESIGN: A prospective, double-blind, randomized, controlled trial of 18 months' duration. SETTING: The study was carried out in 32 health centres (out of a total of approximately 850) in Sweden. SUBJECTS: In all, 681 subjects, aged 30-59 years, were randomized. They had at least two cardiovascular risk factors in addition to moderate primary hyperlipidaemia (total cholesterol of at least 6.50 mmol L-1 on three occasions measured by Reflotron triglycerides less than 4.0 mmol L-1 and an LDL:HDL cholesterol ratio of more than 4.0). Most (87%) of the subjects were males; 626 subjects (92%) completed the 18-month follow-up. INTERVENTION: Half the subjects were randomized to 'intensive advice' given in group sessions led by doctors and nurses in primary care. The other half received 'usual advice'. In each of the two advice groups, one-third received an active lipid-lowering drug (pravastatin), one-third placebo, and one-third no drug at all. The tablets were titrated to achieve a 15% reduction in cholesterol. MAIN OUTCOME MEASURES: Changes in the overall Framingham risk score, and the development of adverse events in each group. RESULTS: The change in Framingham risk score was significantly reduced only in subjects taking lipid-lowering medication (together with intensive advice -0.13; 95% CI-0.20, -0.06, and together with usual advice 0.16; 95% CI -0.23, -0.09). The other subjects receiving intensive advice tended to fare better than those on usual advice. Lifestyle was not influenced significantly over the study period. The ritual of daily medication did not affect the outcome. CONCLUSION: As expected, lipid-lowering medication reduced serum cholesterol as well as overall cardiovascular risk in subjects with several risk factors for cardiovascular disease. There was no additive effect of intensive advice to these subjects. However, there was a meagre but significant effect of intensive advice in subjects not receiving active lipid-lowering drugs. One explanation for this difference may be that those on active lipid-lowering medication who had substantial drops in cholesterol might have felt less inclined to change their lifestyle compared with those on other treatment regimens who had less successful drops in cholesterol. There was no benefit from the ritual of taking daily medication. PMID- 8708587 TI - The cost effectiveness of lipid lowering in Swedish primary health care. The CELL Study Group. AB - OBJECTIVE: To evaluate the cost-effectiveness of two types of advice (usual and intensive) to lower cardiovascular risk, with or without pharmacological medication aimed at lowering cholesterol levels. DESIGN: Prospective, randomized, controlled clinical study of 18 months' duration. SETTING: Thirty-two primary health care centres in Sweden. SUBJECTS: A total of 384 males, aged 30-59 years, with at least one cardiovascular risk factor in addition to moderate primary hyperlipidaemia; of these, 355 completed the 18-month follow-up. INTERVENTIONS: Intensive advice consisted of group sessions led by a health care professional; the usual level of advice was given at follow-up visits. The pharmacological intervention consisted of pravastatin. The goal was to achieve a 15% reduction in cholesterol. MAIN OUTCOME MEASURES: Cost per life-year gained based on the change in serum cholesterol and the net intervention cost of the four treatment options. RESULTS: The usual level of advice and intensive advice in combination with pharmacological treatment achieved no incremental effects and were not considered in the cost-effectiveness analysis. The cost per life-year gained of pharmacological treatment compared with intensive advice decreased. The cost per life-year gained of pharmacological treatment compared with no treatment was about $61,000, if no adverse consequences on noncardiovascular mortality were assumed. CONCLUSIONS: According to the results of the CELL trial, intensive advice is not a cost-effective strategy compared with lipid-lowering drug treatment. However, it is also doubtful whether drug treatment as primary prevention is cost-effective compared with no treatment in the studied patient population. PMID- 8708588 TI - Polyarteritis nodosa: ischaemic intestinal pain successfully treated with nitroglycerin. AB - Inflammation of medium and small sized arteries caused by polyarteritis nodosa (PAN) sometimes causes acute and life threatening ischaemic intestinal complications. We report a case of PAN presenting with severe abdominal pain successfully treated with nitroglycerin. PMID- 8708589 TI - Brain abscesses: the lung connection. AB - Pulmonary arteriovenous fistulas are uncommon abnormalities of capillary development which cause right to left shunting and, if not treated, may lead to severe neurological complications, including meningitis and brain abscess. Pulmonary arteriovenous fistulas are commonly a result of hereditary haemorrhagic telangiectasia (Rendu-Osler-Weber disease) and both conditions may be readily diagnosed by careful history taking and physical examination. Two cases of brain abscess associated with hereditary haemorrhagic telangiectasia, which remained unrecognized for many years, are reported. These cases emphasize the importance of early diagnosis and treatment of pulmonary arteriovenous fistula in preventing central nervous system infections. PMID- 8708590 TI - Idiopathic CD4+ lymphocytopenia and systemic vasculitis. AB - The syndrome defined as "idiopathic CD4 lymphocytopenia' (ICL) is a rare disease of unknown aetiology, often associated with severe depression of immune defences and the occurrence of opportunistic infections. A case is reported wherein a severe immunodeficiency syndrome with persistent idiopathic CD4+ lymphopenia developed in a woman suffering from systemic microscopic polyarteritis; no signs of HIV 1/2 or HTLV I/II infection were evident. The patient died of widespread opportunistic infections. The association of ICL with vasculitis has never been reported until now. A link between the two diseases cannot be ruled out. PMID- 8708591 TI - Systemic lupus erythematosus as a cause of the pulmonary renal syndrome. PMID- 8708592 TI - 'Calcific degeneration of the aortic valve in old age: is the development of flow obstruction predictable?'. PMID- 8708593 TI - Lipoproteins, anticardiolipin autoantibodies and artherosclerosis. PMID- 8708594 TI - Screening for diabetic retinopathy. PMID- 8708595 TI - Should metformin be used to improve blood lipid pattern in nondiabetic patients with coronary heart disease? PMID- 8708596 TI - Serum ferritin and selective iron prophylaxis in pregnancy? PMID- 8708597 TI - Prevention of cardiac sudden death by N-3 fatty acids: a review of the evidence. AB - The essential n-6 and n-3 polyunsaturated fatty acids can prevent ischaemia induced ventricular fibrillation in rats, dogs and marmosets. In isolated neonatal rat cardiac myocytes, these have been shown to prevent tachyarrhythmias caused by elevated calcium concentrations, toxic levels of ouabain, a beta adrenergic agent (isoproterenol), lysophosphatidylcholine and acylcarnitine. The antiarrhythmic effect is caused by a reduction in electrical excitability caused by partitioning of the free polyunsaturated fatty acids into the phospholipid cell membranes of the cardiac myocytes, which modulates membrane ion channels. Two clinical trials suggest they could prevent sudden cardiac death in humans. PMID- 8708598 TI - Atherosclerosis and expression of the neuro-immune-endocrine system. PMID- 8708599 TI - Making doctors change. PMID- 8708600 TI - Attribute conjunctions and the part configuration advantage in object category learning. AB - Five experiments demonstrated that in object category learning people are particularly sensitive to conjunctions of part shapes and relative locations. Participants learned categories defined by a part's shape and color (part-color conjunctions) or by a part's shape and its location relative to another part (part-location conjunctions). The statistical properties of the categories were identical across these conditions, as were the salience of color and relative location. Participants were better at classifying objects defined by part location conjunctions than objects defined by part-color conjunctions. Subsequent experiments revealed that this effect was not due to the specific color manipulation or the role of location per se. These results suggest that the shape bias in object categorization is at least partly due to sensitivity to part location conjunctions and suggest a new processing constraint on category learning. PMID- 8708601 TI - Representational momentum, centripetal force, and curvilinear impetus. AB - In 3 experiments, observers witnessed a target moving along a circular orbit and indicated the location at which the target vanished. The judged vanishing point was displaced forward in the direction of implied momentum and inward in the direction of implied centripetal force. In general, increases in either the angular velocity of the target or the radius length of the orbit increased the magnitude of forward displacement. If both angular velocity and radius length were varied, then increases in either angular velocity or radius length also increased the magnitude of inward displacement. The displacement patterns were consistent with hypotheses that analogues of momentum and centripetal force were incorporated into the representational system. A framework is proposed that accounts for (a) the forward and inward displacements and (b) naive-physics data on the spiral tube problem previously interpreted as suggesting a belief in a naive curvilinear-impetus principle. PMID- 8708602 TI - Summation of activation: evidence from multiple primes that converge and diverge within semantic memory. AB - Six experiments addressed the combinatorial influence of multiple related primes in naming, lexical decision and relatedness judgment performance. Primes either converged on a single semantic representation (e.g., LION-STRIPES-TIGER) or diverged onto distinct semantic representations (e.g., KIDNEY-PIANO-ORGAN). The facilitatory influence of 2 related primes was well predicted by the sum of the influences from the single-related-prime conditions (a) for both convergent and divergent primes, (b) in lexical-decision and naming, (c) across varying prime target stimulus onset asynchronies, and (d) under target-degradation conditions that increased the priming effects. The relatedness-judgment task yielded an additive pattern of priming for convergent prime conditions; however, an underadditive pattern of priming was found for divergent prime conditions. Discussion focuses on the role of attentional systems that modulate the type of information used to perform a given task. PMID- 8708603 TI - Processing across the language boundary: a cross-modal priming study of Spanish English bilinguals. AB - A cross-modal naming paradigm was used to assess within- and between-language sentential priming in Spanish-English bilinguals. The paradigm used single language auditory texts with visual target words under normal, visually degraded, speeded, and delayed naming conditions. Cross-language priming was always observed when the target language was predictable (in the blocked condition), even under speeded conditions. When the target language was unpredictable (in the mixed condition), cross-language priming was observed only when response was delayed (delayed naming) and under a subset of conditions when word recognition was delayed (visual degradation). Results are compatible with the idea that cross language priming in a sentence context is more likely to involve the use of expectations, strategic processes, or both that allow bilinguals to tune themselves to external conditions. There are enough exceptions to the general rule, however, to warrant a reconsideration of the lexical-postlexical dichotomy. Implication for modular versus interactive models of lexical access are discussed. PMID- 8708604 TI - Pronoun resolution without pronouns: some consequences of memory-based text processing. AB - A memory-based processing approach to discourse comprehension emphasizes the rapid deployment of information in memory to facilitate understanding of the text that is currently being read. S. B. Greene, R. J. Gerrig, G. McKoon, and R. Ratcliff (1994) demonstrated that when a text described the reunion of 2 characters who had previously discussed a 3rd character, the accessibility of the 3rd character increased, and the use of an unheralded pronoun (R. J. Gerrig, 1986) to refer to that character was felicitous. In experiments in this article, the authors demonstrate that concepts related to the unheralded pronoun also increase in accessibility and that those concepts form associations in memory with concepts present in the discourse at the time the pronoun is used. The authors also show that the increase in accessibility for the referent of the pronoun, as well as the appropriate long-term memory associations, occurs even in the absence of the pronoun. PMID- 8708605 TI - Level-of-processing effects in word-completion priming: a neuropsychological study. AB - Recent reviews (A.S. Brown & D.B. Mitchell, 1994; B. Challis & D.R. Brodbeck, 1992) concluded that level-of-processing (LOP) manipulations affect priming in perceptual tasks, contrary to earlier suggestions that such tasks are insensitive to LOP. In 3 experiments with amnesic patients and control subjects, the authors examined the effect of LOP manipulations on priming in word-stem and word fragment completion and on recognition memory. Amnesic patients exhibited reduced or near-zero LOP effects in word-completion priming compared with control subjects. LOP affected recognition memory for both amnesic patients and control subjects, confirming that the LOP manipulation affected explicit memory. When the effect of explicit retrieval on control performance was reduced by using a low level encoding task, priming was the same for amnesic patients and control subjects. The authors suggest that LOP effects in word-completion priming tasks reflect the influence of explicit retrieval, which can be used usefully by control subjects but much less so by amnesic patients. PMID- 8708606 TI - Explicit and implicit learning of event sequences: evidence from event-related brain potentials. AB - Event-related brain potentials (ERPs) were recorded during a serial reaction time (RT) task, where single deviant items seldom (Experiment 1) or frequently (Experiment 2) replaced 1 item of a repeatedly presented 10-item standard sequence. Acquisition of sequence knowledge was reflected in faster RTs for standard as compared with deviant items and in an enhanced negativity (N2 component) of the ERP for deviant items. Effects were larger for participants showing explicit knowledge in their verbal reports and in a recognition test. The lateralized readiness potential indicated that correct responses were activated with shorter latencies after training. For deviant items, participants with explicit knowledge showed an initial activation of the incorrect but expected response. These findings suggest that the acquisition of explicit and implicit knowledge is reflected in different electrophysiological correlates and that sequence learning may involve the anticipatory preparation of responses. PMID- 8708607 TI - Alignment-based nonmonotonicities in similarity. AB - According to the assumption of monotonicity in similarity judgments, adding a shared feature in common to 2 items should never decrease their similarity. Violations of monotonicity are not predicted by feature- or dimension-based models but can be accommodated by alignment-based models in which the parts of one compared display are placed in correspondence with the parts of the other display. In 2 experiments, evidence for nonmonotonicities is obtained that is generally consistent with the alignment-based model SIAM (similarity as interactive activation and mapping; R.L. Goldstone, 1994). The calculation of similarity in this model involves an interactive activation process whereby correspondences between the parts of compared displays mutually and concurrently influence each other. As SIAM predicts, the occurrence of nonmonotonicities depends on perceptual similarity of features and the duration of presented comparison. PMID- 8708608 TI - Medical student teaching and family physicians' use of time. PMID- 8708609 TI - Medical student teaching and family physicians' use of time. PMID- 8708610 TI - Medical student teaching and family physicians' use of time. PMID- 8708611 TI - Predicting sinusitis. PMID- 8708612 TI - Endocervical cells on Pap smears. PMID- 8708613 TI - Calcium supplementation during pregnancy. PMID- 8708615 TI - Iron deficiency anemia after negative GI evaluation. PMID- 8708614 TI - Effect of beta-blockade on chronic heart failure. PMID- 8708616 TI - Using serology to detect H pylori infection. PMID- 8708617 TI - Membrane stripping to promote labor. PMID- 8708618 TI - Effect of reduced dietary sodium on blood pressure. PMID- 8708619 TI - The value of computers in the clinic. PMID- 8708620 TI - Cost-effectiveness of the nicotine patch. PMID- 8708621 TI - Diagnosis and treatment of tinea versicolor. AB - Tinea versicolor (pityriasis versicolor) is a common superficial fungal infection of the stratum corneum. Caused by the fungus Malassezia furfur, this chronically recurring disease is most prevalent in the tropics but is also common in temperate climates. Treatments are available and cure rates are high, although recurrences are common. Traditional topical agents such as selenium sulfide are effective, but recurrence following treatment with these agents is likely and often rapid. Currently, therapeutic interest is focused on synthetic "-azole" antifungal drugs, which interfere with the sterol metabolism of the infectious agent. Ketoconazole, an imidazole, has been used for years both orally and topically with great success, although it has not been approved by the Food and Drug Administration for the indication of tinea versicolor. Newer derivatives, such as fluconazole and itraconazole, have recently been introduced. Side effects associated with these triazoles tend to be minor and low in incidence. Except for ketoconazole, oral antifungals carry a low risk of hepatotoxicity. PMID- 8708622 TI - Managed care contracting for physicians. PMID- 8708623 TI - Answering clinical questions. AB - BACKGROUND: Physicians often have unanswered clinical questions. The purpose of this study was to determine how often the answers to these questions can be found in the medical literature. METHODS: We collected unanswered clinical questions from family physicians at the end of clinical half-days. The authors and medical librarians then used textbooks and MEDLINE to find answers to each question. We returned to the physicians one to five selected references for each question. Each physician rated these sources on how well they answered the questions and how they might influence the physician's practice. RESULTS: One hundred three questions were gathered. Physicians asked an average of 0.5 questions per half day. We searched for answers to 86 questions, and the physicians returned ratings for 84. Forty-five (54%) of these questions were fully or nearly fully answered by the materials returned to the physicians. Of the questions for which answers were found, MEDLINE searches accounted for 71%; textbooks, 20%; and a combination, 9%. MEDLINE searches took an average of 27 minutes, whereas textbook searches averaged 6 minutes. CONCLUSIONS: The medical literature can provide answers to a majority of clinical questions; however, finding these answers is time-consuming and expensive. Physicians need more efficient ways to answer their clinical questions. PMID- 8708624 TI - Benign fever following vaginal delivery. AB - BACKGROUND: In patients with fever following vaginal delivery, physicians must differentiate benign self-resolving fevers from fevers with more serious causes, especially endometritis. To help differentiate these clinical entities, we explored the characteristics and risk factors for benign "single-day" postpartum fever. METHODS: We conducted a retrospective cohort study of 2137 vaginal deliveries. Patients were randomly selected from the 25,687 vaginal deliveries that took place between 1979 and 1992 at The University of Iowa Hospitals and Clinics. The data were analyzed using odds ratios and multiple logistic regression. RESULTS: Benign fevers occurred in 3.3% of patients, while endometritis was diagnosed in 1.6%. After controlling for confounding variables, two clinical factors were independently associated with single-day fever: primiparity (odds ratio [OR], 3.4; 95% confidence interval [CI], 2.0 to 5.7) and use of a uterine pressure catheter (OR, 2.4; 95% CI, 1.5 to 3.7). These factors were not associated with endometritis. The first postpartum temperature elevation ( > or = 38.0 degrees C) occurred earlier in patients with single-day fever than in patients with endometritis (4.0 +/- 4.6 hours postpartum vs 30.2 +/- 27.0 hours postpartum, P < .001). The maximum temperature elevation was lower, on average, in patients with single-day fever than in patients with endometritis (38.2 degrees +/- 0.2 degrees C vs 38.9 degrees +/- 0.6 degrees C, P < .001). CONCLUSIONS: Single-day fever was more likely to occur in primiparous women and in women who were monitored with a uterine pressure catheter. Most women with benign single-day fevers had early low-grade fevers, whereas women with endometritis had higher fevers that occurred later in the postpartum period. PMID- 8708625 TI - Substance abuse among patients with chronic back pain. AB - BACKGROUND: Little is known about substance use and substance use disorders among primary care patients with chronic back pain. This study compared groups of patients with and without chronic back pain for the prevalence of substance use and substance use disorders. It also assessed the temporal relationship between the onset of chronic back pain and that of substance use disorders. METHODS: Adult patients aged 18 to 59 years who made three or more visits for back pain to a family practice clinic were eligible for the pain group. The comparison group consisted of a random sample of patients of the same ages who made appointments with the same clinic. A validated diagnostic interview about substance use disorders and other issues related to substance use was administered. RESULTS: Ninety-two percent of the patients in the chronic pain group reported severe pain, high disability, and severe to moderate limitation of activity. Two thirds (67%) of this group reported having continuous pain, and 21% experienced at least one episode of pain daily. Forty-four percent said their pain continously interfered with their activities, and 31% reported daily disruption of activity. There was little difference, however, in the adjusted rates of lifetime and current substance use disorders between the chronic pain and comparison groups. Lifetime prevalence rates were 54% for the pain group and 52% for the comparison group; current prevalence rates were 23% for both the pain and comparison groups. Substance abuse preceded the onset of pain by as much as 20 years for 77% of patients with chronic pain who had current substance use disorders and 63% of those who had lifetime substance use disorders. CONCLUSIONS: Chronic back pain did not connote special risk for current substance use disorders. PMID- 8708626 TI - Factors related to planned and unplanned pregnancies. AB - BACKGROUND: Given the efficacy of most contraceptive options, it is of concern that most pregnancies in the United States are unplanned. Besides reducing the woman's and family's preparedness for parenting, unplanned pregnancies are at higher risk for inadequate prenatal care, perinatal morbidity, and significant postnatal problems. Little is known about the factors responsible for the high rates of unplanned pregnancy. METHODS: One hundred ten pregnant women were surveyed to examine factors relating contraception to unplanned pregnancy. RESULTS: Sixty-five percent of pregnancies were unplanned. There was a statistically significant association between having unplanned pregnancies and being single or divorced. Women who had planned their pregnancies tended to be more satisfied with contraceptives. In sexual encounters, women with unplanned pregnancies were more likely to use no contraception or to practice "withdrawal" or use condoms rather than hormonal contraception; to be influenced by their partner regarding birth control use; and to forget to use contraception. CONCLUSIONS: All women of childbearing age who are sexually active can benefit from planning pregnancies. Counseling that accesses a woman's expectations regarding birth control, followed by a careful explanation of the side effects of a contraception choice, may reduce the rate of unplanned pregnancy. Counselling the male partner or sexually active men in contraceptive options may be equally important. Understanding factors that result in satisfaction with contraception may reduce unplanned pregnancies. PMID- 8708627 TI - Family medicine research in the community setting: what can we learn from successful researchers? AB - BACKGROUND: There is little information describing family physician researchers who work outside academic medical centers. This report describes the motivating factors and resources used by community residency faculty and nonfaculty family physicians who perform research. METHODS: We sent a questionnaire to community residency faculty and nonfaculty family physicians who published at least one paper in the family medicine literature from 1992 through 1994. The survey focused on previous research experience, training, and collaboration with university colleagues, and included an open-ended question about motivations and obstacles to research. RESULTS: The majority (60%) of community faculty and nonfaculty family physicians surveyed reported previous research experience on the undergraduate, medical school, or residency level. Research training received during residency was evaluated as poor. Sixty-nine percent of the respondents reported being successful at acquiring research funding, and 60% reported receiving funding from foundations. Reported keys to success included mentoring, a supportive infrastructure, and an inherent enjoyment in doing research. These factors did not differ between community residency faculty and nonfaculty physicians. CONCLUSIONS: For community-based family physicians, success at conducting and publishing research is enhanced by the availability of mentoring, support from local or national foundations, and previous research experience. Respondents identified research training during residency as one area that needs improvement. PMID- 8708628 TI - False-negative endoscopic biopsy of colonic adenocarcinoma in a young man. AB - A 26-year-old man presented with intermittent bright red blood per rectum. His physical examination was unremarkable; however, because microcytic anemia was noted, the patient underwent colonoscopy. A large ascending colonic lesion was noted, a biopsy of which was negative for cancer. Nevertheless, the patient underwent a radical hemicolectomy. Adenocarcinoma of the cecum was found. This case report is an example of a false-negative endoscopic-directed biopsy. A review of recorded video-endoscopy assisted the family physician in appropriate management. PMID- 8708629 TI - Atypical glandular cells of undetermined significance and adenocarcinoma in situ: summoning colposcopic expertise? AB - In comparison with cervical squamous neoplasia, glandular cell neoplasia is uncommon. The evaluation of a patient with atypical glandular cells of undetermined significance is challenging because subtle colposcopic signs are frequently inaccessible to view and cytologic interpretations are extremely challenging for many cytopathologists. PMID- 8708630 TI - Home. PMID- 8708631 TI - Developmental regulation of the urea-cycle enzyme arginase in the direct developing frog Eleutherodactylus coqui. AB - Direct developing organisms obviate the larval intermediary from their ontogeny, hatching as miniature adults. To investigate this phenomenon, we have examined the developmental expression of arginase in the direct developing frog Eleutherodactylus coqui. An enzyme in the ornithine-urea cycle, the activation of liver arginase is necessary for the switch from ammonotelism to ureotelism which occurs when many frogs metamorphose and assume a terrestrial existence. Arginase enzyme activity is detectable at low levels in late prehatching stages of E. coqui, and increases at hatching, at which point the protein becomes detectable on Western blots. The activity increases gradually during posthatching development, reaching maximal levels at approximately the same time as yolk resorption is completed. Thyroid hormone is responsible for upregulating arginase activity during metamorphosis in Rana, but the role of thyroid hormone in direct developing frogs is unknown. A high dose (250 nM) of the thyroid hormone analogue 3,3'5-triiodo-L-thyronine (T3) caused precocious induction of arginase protein and activity, showing that even in a direct developing frog, some level of responsiveness to the metamorphic trigger, thyroid hormone, has been retained. PMID- 8708632 TI - Comparison of the body temperature rhythms of diurnal and nocturnal rodents. AB - The daily rhythm of body temperature of six rodent species, three of them nocturnal (Rattus norvegicus, Mesocricetus auratus, and Meriones unguiculatus) and three diurnal (Octodon degus, Spermophilus tridecemlineatus, and S. richardsoni), was studied by telemetry under a 14L:10D light-dark cycle. Significant interspecies differences were found in the mean level (range: 36.0 37.4 degrees C), amplitude (range: 2.1 degrees-4.3 degrees C), robustness (range: 115-185 Qp), and acrophase (range: 8.5-16.0 hr) of the temperature rhythm. In addition, a positive correlation (r = 0.79) was found between body mass and robustness of the rhythm across species. However, the only consistent difference between the diurnal and nocturnal species was the acrophase, which occurred during the day in the diurnal species (range: 8.5-11.5 hr) and during the night in the nocturnal species (range: 13.5-16.0 hr). PMID- 8708633 TI - Separating the effects of alertness from the effects of encoding in a pitch imagery task. AB - The facilitatory effects of auditory imagery have often been attributed to priming. However, the facilitatory effects of a prime on subsequent processing can be separated into effects related to specific perceptual processes, such as encoding, and effects related to more general processes, such as alertness. Whereas research on auditory imagery has focused on code-activated facilitation, the role of alertness in image-induced facilitation has been largely presumed rather than empirically investigated. Therefore, a study was designed to isolate the facilitatory effects of alertness from those of encoding. The results show that the nonspecific alerting effects of auditory imaging were quite extensive--a nearly 78-ms decrease in reaction time--compared with the effects of encoding. PMID- 8708634 TI - Asymmetrical perception of changing intensity in short tonal stimuli: duration of stimulus. AB - A number of reports have suggested that changing intensity in short tonal stimuli is asymmetrically perceived. In particular, steady stimuli may be heard as growing louder; stimuli must decrease in intensity to be heard as steady in loudness. The influence of stimulus duration on this perceptual asymmetry was examined. Three participants heard diotic tonal stimuli of eight durations between 0.8 s and 2.5 s. Each stimulus increased, decreased, or remained steady in intensity; initial intensity was 40 dB SPL (sound pressure level relative to 0.0002 dynes/cm2), and carrier frequency was 1 kHz. Participants made forced binary responses of "growing louder" or "growing softer" to each stimulus. For each duration, that value of intensity change eliciting equal numbers of both responses was determined. The results indicated a pronounced perceptual asymmetry for 0.8-s stimuli, which diminished for longer stimuli; changing intensity in 2.5 s stimuli was perceived symmetrically. Additionally, sensitivity to changing intensity improved as stimulus duration increased, suggesting that responses may be based in part on the difference in intensity between the beginning and end of the stimulus. Possible ramifications of the asymmetry reside in (a) the percussive nature of many natural sounds and (b) selective responding to approaching sound sources. PMID- 8708635 TI - Visual and auditory information as determinants of primacy effects. AB - Recent research has demonstrated that auditory and visual information have similar mnemonic properties. The relative influence of verbal, auditory, and visual stimulus materials on the primacy effect in free recall was studied. Significant primacy effects were obtained when either verbal or auditory-input stimuli were presented. However, primacy effects were suppressed when pictorial materials were used, principally because of elevation of recall for midlist and later elements. Instructions to use visual mental imagery at encoding suppressed primacy effects for verbal materials, but not for auditory materials, indicating that although auditory information processing is probably similar to phonological processing within working memory, auditory processing may share cognitive resources with visuospatial processing. Results are shown to be predictable and interpretable within the item-specific/relational information distinction of Hunt and Einstein (e.g., 1981), and within the working-memory theory of Baddeley (e.g., 1986). PMID- 8708636 TI - Serial pattern learning in senescent rats: a comparison of acquisition performance with young and hippocampal-lesioned rats. AB - The acquisition of a serial pattern was assessed in young (3-month), senescent (24-month), and hippocampal-lesioned rats. All animals were trained for 25 days on a four-element, nonmonotonic pattern consisting of 14, 0, 3, and 7 food pellets, respectively. Young rats were capable of distinguishing among the elements of the series, with the exception of the 3- and 7-pellet elements, as indexed by running times. Conversely, the senescent rats were largely incapable of patterned responding, and the tracking performance of the hippocampal-lesioned rats was intermediate between those of the young and old rats. Results are discussed in terms of the possible changes that accompany aging in rodents, including age-related changes in the hippocampal system. PMID- 8708637 TI - Sex and sociocultural correlates of urinary incontinence in Israeli preschool children. AB - Urinary incontinence in children aged 2 to 6 years whose mothers varied in ethnic descent, in level of education, and in immigration status was studied in Israel. Mothers of 302 children were interviewed in well-baby clinics regarding primary and secondary urinary incontinence of their children. A significant age trend was found for attaining primary urinary continence. However, various interactions between mothers' ethnic descent, mothers' level of education, and mothers' immigration status were associated with different rates of primary and secondary urinary incontinence in their children. Moreover, sex differences in rates of primary and secondary urinary incontinence were also associated with mothers' ethnic descent and level of education. It is suggested that different levels of maternal emotional availability to the child in the various subgroups can account for most of the findings. PMID- 8708638 TI - Evaluating neurological outcome measures: the bare essentials. PMID- 8708640 TI - Subarachnoid haemorrhage. PMID- 8708639 TI - Multimodal monitoring in neurointensive care. PMID- 8708641 TI - Incidence, management, and outcome of post-traumatic syringomyelia. In memory of Mr Bernard Williams. AB - OBJECTIVE: To determine the incidence of clinically diagnosable post-traumatic syringomyelia (PTS). METHODS: A population of 815 consecutive patients with traumatic spinal cord injuries was studied between January 1990 and December 1992. RESULTS: Reviews of all records, full clinical evaluation, and thorough neurological examination of all patients disclosed 28 patients in whom PTS was confirmed radiologically (3.43%). The incidence of the presenting symptoms, including bladder dysfunction, is described. The level and density of cord lesion was correlated with incidence and it was found that posttraumatic syringomyelia was twice as common in patients with complete injuries than in patients with incomplete injuries. The highest incidence was found in patients with complete dorsal and complete dorsolumbar injuries. The interval between injury and diagnosis ranged from six months to 34 years (mean 8.6 years). This interval was shortest in patients with complete dorsal and incomplete cervical and dorsolumbar cord injuries. CONCLUSIONS: Reduction of the size of the syrinx seen on postoperative MRI correlated well with a satisfactory clinical outcome in 85% of patients. PMID- 8708642 TI - Avoiding false positive diagnoses of motor neuron disease: lessons from the Scottish Motor Neuron Disease Register. AB - OBJECTIVES: To describe the frequency and characteristics of those patients initially registered with the Scottish Motor Neuron Disease Register (SMNDR) but who subsequently had a diagnosis other than MND made (false positives), to analyse the features which led to a revised diagnosis, and to draw conclusions which might improve routine neurological practice. METHODS: The Scottish Motor Neuron Disease Register is a community based, prospective disease register to identify and follow up all incident cases of motor neuron disease in Scotland. Fifty three patients out of a total of 552 registered are presented, who, after initial registration, were later excluded because they failed to satisfy the register's diagnostic criteria. RESULTS: Seven of these patients were labelled as "MND plus" syndromes and may represent a distinct subset of MND. The remaining 46 patients had an alternative diagnosis made (false positive group), accounting for 8% of the total. In half of these cases, potentially beneficial therapies are available. The predominant reasons which lead to a diagnostic revision were: failure of symptom progression, development of atypical clinical features for MND, and investigation results. CONCLUSIONS: Patients with MND should undergo thorough and relevant investigations at presentation with the emphasis on neuroradiological imaging and neurophysiology; all patients should be followed up by an experienced neurologist, particularly those in whom symptoms and signs are restricted to either the bulbar or spinal muscles; failure of symptom progression or development of atypical features should lead to an early reassessment; finally, patients should be informed of the diagnosis only when it is secure. PMID- 8708643 TI - Historical note on the therapeutic use of botulinum toxin in neurological disorders. PMID- 8708644 TI - Observer variation in the interpretation of intra-arterial angiograms and the risk of inappropriate decisions about carotid endarterectomy. AB - OBJECTIVE: To determine how often observer variation in the interpretation of intra-arterial angiograms might alter the decision whether or not to refer patients for carotid surgery. METHODS: A prospective study was carried out in a consecutive series of 99 patients with transient ischaemic attacks and minor strokes. Interpretable angiographic films were available for 179 carotid artery bifurcations. Stenosis of the internal carotid artery was measured using mm scales, independently by three different radiologists (A, B, and C), using the European Carotid Surgery Trial method. RESULTS: An analysis of the grouped data showed good to moderate agreement by kappa statistics for radiologists A v B, B v C, and A v C of 0.68, 0.60, and 0.70 respectively. The mean absolute difference in the estimate of stenosis by each of the different radiologists (interobserver variation) was 9.5% and for each radiologist on two separate occasions (intraobserver variation) 8.4%. The degree of observer error was smallest among severely stenosed arteries. Although the absolute differences were small, "clinically important" differences which could change the treatment recommended from surgery to no surgery (or vice versa) occurred between radiologists A and B, B and C, and A and C in: seven (3.9%), six (3.4%), and 11 (6.1%) vessels respectively. CONCLUSIONS: Because observer variation affects all of the imaging methods (Doppler, duplex, contrast arteriography, and MR angiography) used to select patients with transient ischaemic attack and stroke, these findings are likely to be widely relevant. Any centre assessing patients with cerebrovascular disease will need to implement strict quality control measures in the interpretation of angiograms (and other vascular imaging procedures) to minimise observer error and thereby reduce the number of inappropriate decisions made to refer for carotid artery surgery or not. PMID- 8708646 TI - Baron Constantin von Economo and encephalitis lethargica. PMID- 8708645 TI - Comparison of positron emission tomography, cognition, and brain volume in Alzheimer's disease with and without severe abnormalities of white matter. AB - OBJECTIVES: To examine cerebral metabolism, cognitive performance, and brain volumes in healthy controls and two groups of patients with probable Alzheimer's disease, one group with severe abnormalities of white matter (DAT+) and the other group with none, or minimal abnormalities (DAT-). METHODS: Neuropsychological tests, CT, MRI, quantitative MRI, and PET studies were carried out to allow comparison between the DAT+ and DAT- groups and the healthy controls. RESULTS: Compared with the healthy controls, both demented groups had significantly reduced global and regional cerebral metabolism, significant brain atrophy, and significantly lower scores on neuropsychological testing. The DAT- patient group showed a pattern of parietal-temporal cerebral metabolic reductions and neuropsychological performance deficits typical of Alzheimer's disease. In addition, metabolism in the association neocortex (AD ratio) and measures of neuropsychological task performance were significantly correlated in the DAT- patient group. Comparison of DAT+ with DAT- patients showed a significantly higher ratio of parietal to whole brain glucose utilisation for the DAT+ group. Moreover, when comparing group z score differences from the healthy controls, the DAT+ group had, on average, smaller differences from controls in the frontal, parietal, and temporal regions than did the DAT- group. Discriminant analysis using metabolic ratios of the frontal, parietal, and temporal regions showed cerebral metabolic patterns to be significantly different among the DAT+, the DAT , and the healthy controls. These differences were due primarily to relatively higher frontal, parietal, and temporal metabolic ratios in the DAT+ group which resulted in discriminant scores for the DAT+ group between the healthy controls and the DAT- group. Group mean scores on tests of neuropsychological performance were not significantly different between the DAT- and DAT+ patients. By contrast with the DAT- group, however, no significant correlations between the AD ratio and any neuropsychological task were seen in the DAT+ group. Multiple regression analysis showed significant between group differences in the relation between the AD ratio and neuropsychological scores on three tasks. The slopes of the relations between the AD ratio and memory scores (memory and freedom from distractability deviation quotient of the Wechsler adult intelligence scale (WMDQ)) also were significantly different for the two groups. CONCLUSIONS: Although multiple causes for abnormalities of white matter exist in patients with Alzheimer's disease, these data suggest that the presence of severe abnormalities of white matter indicate a second pathological process in the DAT+ patients. The DAT- patients showed the parietal-temporal metabolic deficits and correlations between association neocortical metabolism and neuropsychological task performance typical of patients with Alzheimer's disease. By contrast, the DAT+ group had a pattern of cerebral metabolism significantly different from healthy controls and DAT+ patients, as well as no significant correlations between metabolism in the association neocortex and neuropsychological performance. These differences probably reflect the superimposed pathology of the abnormalities of white matter which may exert their affect through disruption of long corticocortical pathways. PMID- 8708647 TI - Lasting reduction of severe spasticity after ending chronic treatment with intrathecal baclofen. AB - OBJECTIVE: To investigate whether the dose of intrathecal baclofen necessary for a sufficient reduction of muscle tone and spasms changes during treatment of severe spasticity. METHODS: A group of 27 patients received intrathecal baclofen for 61 (SD 18) months. RESULTS: Spasticity remained absent or strongly reduced after stopping the intrathecal baclofen infusion in seven patients. The dose of baclofen could be reduced to 40% of that dose which was originally necessary in 10 patients. The dose remained the same or increased slightly in 10 patients. Possible reasons for the continuing reduction of spasticity after terminating long term intrathecal baclofen infusion in some patients could be: lasting morphological changes in spinal cord neurons by second messenger controlled modulation of gene expression, a toxic effect of baclofen on spinal neurons, muscular atrophy, inflammation due to the catheter, or progression of multiple sclerosis. CONCLUSIONS: A higher initial daily dose of intrathecal baclofen might lead to a faster, lasting suppression of spasticity and the development of spastic symptoms might even be prevented by pre-emptive treatment with baclofen in patients with newly acquired lesions of the spinal cord. PMID- 8708648 TI - Herpes encephalitis is a disease of middle aged and elderly people: polymerase chain reaction for detection of herpes simplex virus in the CSF of 516 patients with encephalitis. The Study Group. AB - OBJECTIVE: To assess the diagnostic potential of the polymerase chain reaction (PCR) in herpes simplex virus (HSV) encephalitis. METHODS: Samples of CSF from 516 patients with encephalitis were studied for HSV-DNA by PCR. RESULTS: Samples taken one to 29 days from the onset of symptoms from 38 patients (7.4%) were positive, 32 (6.2%) for HSV-1 and six (1.2%) for HSV-2. At follow up, eight of 28 patients studied were still HSV-PCR positive. A diagnostic serum:CSF antibody ratio to HSV but not to other viruses was detected in 25 of the 38 HSV-PCR positive patients thus supporting the initial PCR findings. Patients positive by HSV-PCR were concentrated in the age group > or = 40 years, and especially in patients aged 60-64 years, of whom nine of 24 (37.5%) were positive. The HSV-PCR was negative in all other patients with encephalitis of known or unknown aetiology. This group included 34 patients with a diagnostic serum:CSF antibody ratio to other viruses. A dual infection, HSV and another microbe, was considered possible in seven patients. CONCLUSIONS: The HSV-PCR is a rapid and useful diagnostic method during the early phase of encephalitis. It may be useful in monitoring the efficacy of treatment and allowing the recognition of new features in the appearance of herpes encephalitis. The HSV-PCR test and antibody determinations from serum and CSF are complementary methods, which should both be applied in pursuit of clinical laboratory diagnosis of these conditions. PMID- 8708649 TI - Saccade disturbances after bilateral lentiform nucleus lesions in humans. AB - OBJECTIVE: To determine the roles of the putamen and pallidum in ocular motor control. METHODS: Eye movements were recorded electro-oculographically in nine patients with bilateral focal lesions affecting the lentiform nucleus, and in 12 age matched control subjects. Reflexive visually guided saccades (gap task), antisaccades, memorised sequences of saccades, memory guided saccades (with visual input only, and with both visual and vestibular inputs), and predictive saccades (with and without gap) were studied. RESULTS: Latency and accuracy of visually guided saccades were normal. The percentage of errors in the antisaccade task and latency of correct antisaccades did not differ significantly from the results of controls. The percentage of errors in saccade sequences was significantly increased. Accuracy of the two types of memory guided saccades was impaired bilaterally. The percentage of predictive saccades was significantly decreased when a gap existed, but unchanged without a gap, compared with controls. Therefore, saccades made immediately in response to an external target (reflexive visually guided saccades and antisaccades) were performed without difficulty, whereas those requiring an internal representation of such a target (such as memory guided saccades, predictive saccades, and saccade sequences) were performed with significant disturbances. CONCLUSIONS: The lentiform nucleus influences the cortical areas involved in the control of saccades when the experimental paradigm requires the use of an internal representation of the target for correct planning and execution of the ensuing saccade. PMID- 8708650 TI - Treatment of chronic inflammatory demyelinating polyneuropathy with cyclosporin A. AB - Chronic inflammatory demyelinating polyneuropathy (CIDP) is an immune mediated polyneuropathy for which there are effective therapies. However, not all patients improve with these treatments. Eight patients with CIDP, two with IgG monoclonal gammopathies, were treated with cyclosporin-A (3 to 5 mg/kg/day). In three, this treatment was successful. It was unsuccessful in four patients who were resistant to other treatments and in one who had initially received cyclosporin-A. There were no serious side effects. In selected patients with CIDP, cyclosporin-A is a potentially useful treatment. PMID- 8708651 TI - Chronic inflammatory demyelinating polyneuropathy associated with carcinoma. AB - The association of chronic inflammatory demyelinating polyneuropathy (CIDP) and carcinoma has rarely been reported and its relevance is debated. Thirty three consecutive patients with probable or definite CIDP (idiopathic or associated with M protein) were investigated. Three patients with definite CIDP had a concomitant carcinoma. One had an IgM paraprotein. Steroids and intravenous immunoglobulins were effective. PMID- 8708652 TI - Respiratory electrophysiological studies in Guillain-Barre syndrome. AB - Respiratory failure is a common and potentially life threatening complication in patients with Guillain-Barre syndrome. The incidence of phrenic nerve involvement and the predictive value of phrenic nerve conduction and diaphragmatic needle EMG were studied in 40 patients with Guillain-Barre syndrome within the first three days of admission to hospital. The negative peak onset latency of the diaphragmatic compound muscle action potential (CMAP), and its amplitude, duration, and area were abnormal in 83%. The need for ventilation was correlated with diaphragmatic CMAP amplitude (P = 0.005), and area (P = 0.001), but not with latency or duration. Abnormalities in diaphragmatic needle EMG were found in 45%, mainly a decreased number of motor unit potentials. The abnormalities correlated with the need for ventilation (P = 0.013). Of the 40% who required ventilation, all had either abnormal phrenic conduction, abnormal diaphragmatic needle EMG, or both. Eighty one per cent of the ventilated patients had abnormal forced vital capacity on the day of the electrophysiological examination. The results indicate that phrenic nerve conduction studies and diaphragmatic EMG are useful in detecting respiratory involvement in patients with Guillain-Barre syndrome and in identifying those at risk of respiratory failure. PMID- 8708653 TI - Autosomal dominant optic atrophy with asymptomatic peripheral neuropathy. AB - The association between hereditary motor and sensory neuropathy (HMSN) and optic atrophy has been termed HMSN type VI. The autosomal dominant inheritance of this syndrome is reported. Three generations were affected with optic atrophy, which differed in some respects from classic dominant optic atrophy, and an asymptomatic, mainly sensory, neuropathy. PMID- 8708655 TI - Intracranial intraparenchymal schwannomas: a series of eight cases. AB - Intraparenchymal schwannomas of the CNS are extremely rare. Between 1979 and 1993 400 cases of intracranial schwannomas were operated on and among them were eight patients with intraparenchymal schwannomas comprising 2% of intracranial benign nerve sheath tumours. Four of them were located in the cerebral hemispheres and two each in the brain stem and in the cerebellum. In two cases, there was associated neurofibromatosis (NF-1 and NF-2, one case each). The age ranged from 6 months to 45 years with a male/female ratio of 3:1 and, surprisingly, six of them were in the left cerebral or cerebellar hemisphere. PMID- 8708654 TI - Aspirin at any dose above 30 mg offers only modest protection after cerebral ischaemia. AB - There is continuing debate about the relative efficacy of low (< 100 mg per day), medium (300 to 325 mg per day), and high (> 900 mg per day) doses of aspirin in patients after a transient ischaemic attack or non-disabling stroke. The purpose of this study was to resolve the issue. Thus a minimeta-analysis was performed on data from 10 randomised trials of aspirin only v control treatment in 6171 patients after a transient ischaemic attack or nondisabling stroke. The data on the trials were listed in an appendix of the report on the second cycle of the Antiplatelet Trialists' Collaboration. There was virtually no difference in relative risk reduction for low, medium, and high doses of aspirin (13%, 9%, and 14% respectively). This equivalence corresponds with the results of the UK-TIA trial in a direct comparison of 300 and 1200 mg. The Dutch TIA trial showed no difference in efficacy of 30 and 283 mg. It is concluded that aspirin at any dose above 30 mg daily prevents 13% (95% confidence interval 4-21) of vascular events and that there is a need for more efficacious drugs. PMID- 8708656 TI - Sex prevalence of focal dystonias. AB - The sex prevalence of idiopathic focal dystonia is reported from a data base review of all patients seen at the National Hospital of Neurology, Queen Square and King's College, London up to 1993. There was a higher prevalence of females to males in all categories of focal dystonia involving the craniocervical region. The female to male ratio for cranial dystonia was 1.92:1 (P < 0.01) and 1.6:1 (P < 0.001) for spasmodic torticollis. On the other hand, twice as many men than women had writer's cramp (M:F = 2.0:1, P < 0.01). At present, there is no clear explanation to account for this differences in the sex prevalence of different types of focal dystonia. PMID- 8708657 TI - Huntington's disease: a molecular genetic and CT comparison. AB - Huntington's disease is a progressive neurodegenerative disease in which the molecular abnormality has recently been described. Before the availability of this molecular marker diagnosis depended on clinical findings, supported in some instances by neuroimaging using CT or MRI. The imaging modalities may show atrophy of the heads of caudate nuclei in affected people. An attempt was made to validate these imaging findings using the molecular test as "gold standard." Retrospective analysis of cranial CT in 16 patients with Huntington's disease and 16 age and sex matched controls was performed. There was a highly significant difference in caudate head size (P < 0.00001) between patients with Huntington's disease and control subjects. However, the sensitivity of diagnosis based on radiological examination alone was only 87.5% in this study. Thus the sensitivity of CT is insufficient to justify its routine use in the investigation of suspected Huntington's disease, unless genetic tests are negative and other diagnoses need to be excluded. PMID- 8708658 TI - A Brazilian cohort of patients with Tourette's syndrome. AB - The clinical features of 32 patients (24 males) with Tourette's syndrome in Brazil were studied. The mean age at onset was 7.1 years, tics being the first symptom in 71% and hyperactivity in 29%. Blinking, grimacing, and shoulder elevation were the most common motor tics and sniffing, throat clearing, and grunting noises, the most frequent vocal tics. Coprolalia was present in 28%, echolalia in 16%, palilalia in 9%, and copropraxia in 25% of patients. Attention deficit and hyperactivity disorder was diagnosed in 63%, and obsessive compulsive behaviour in 44% of patients. In 84% of patients there was a family history of tics whereas attention deficit and hyperactivity disorder and obsessive compulsive behaviour were respectively present in relatives of 19% and 53% of the patients studied. These data suggest that Tourette's syndrome in Brazil is not clinically different from other countries, supporting the notion that genetic factors play the most important part in its aetiology. PMID- 8708659 TI - Neuroleptic induced parkinsonism: MRI findings in relation to clinical course after withdrawal of neuroleptic drugs. AB - Parkinsonism is a common complication of neuroleptic drug use; however, the pathophysiology of the persistence of parkinsonian symptoms after withdrawal of neuroleptic drugs is poorly understood. Twenty patients with neuroleptic induced parkinsonism were studied by high field MRI. Persistence of symptoms was associated with different findings depending on the age of the patients--namely, putaminal hypointensity in young patients and striatal hyperintensities in old patients. High field MRI may be useful in identifying patients at higher risk for neuroleptic induced parkinsonism. PMID- 8708660 TI - Optokinetic stimulation influences the disturbed perception of body orientation in spatial neglect. AB - The effect of optokinetic stimulation on the disturbed perception of body orientation in three patients with right brain damage and spatial neglect was examined. The patients were asked to direct a laser point to the position which they felt lay exactly "straight ahead" of their bodies' orientation. Without stimulation they localised the body's sagittal midplane markedly to the right of the objective orientation. The patients' horizontal displacement of the sagittal midplane was reduced by a movement of the surround to the left and worsened by a movement to the right. The findings are consistent with those found in patients with spatial neglect using vestibular and neck proprioceptive stimulation. They show that visual input, together with vestibular and neck proprioceptive input, is used for computing a central representation of egocentric space. In spatial neglect this coordinate transformation works with a systematic error and deviation of the spatial reference frame to the ipsilesional side. The positive effect of optokinetic stimulation in patients with spatial neglect is interpreted with a "correction" of the neural coordinate transformation process by producing asymmetric input at the sensory organs of the contributing channels. PMID- 8708661 TI - Cobalt-55 positron emission tomography in traumatic brain injury: a pilot study. AB - Traumatic brain injury is usually assessed with the Glasgow coma scale (GCS), CT, or MRI. After such injury, the injured brain tissue is characterised by calcium mediated neuronal damage and inflammation. Positron emission tomography with the isotope cobalt-55 (Co-PET) as a calcium tracer enables imaging of affected tissue in traumatic brain injury. The aim was to determine whether additional information can be gained by Co-PET in the diagnosis of moderate traumatic brain injury and to assess any prognostic value of Co-PET. Five patients with recent moderately severe traumatic brain injury were studied. CT was performed on the day of admission, EEG within one week, and MRI and Co-PET within four weeks of injury. Clinical assessment included neurological examination, GCS, neuropsychological testing, and Glasgow outcome scale (GOS) after one year. Co PET showed focal uptake that extended beyond the morphological abnormalities shown by MRI and CT, in brain regions that were actually diagnosed with EEG. Thus Co-PET is potentially useful for diagnostic localisation of both structural and functional abnormalities in moderate traumatic brain injury. PMID- 8708662 TI - Thymus serpyllum (wild thyme). PMID- 8708663 TI - Progressive ataxia, focal seizures, and malabsorption syndrome in a 41 year old woman. PMID- 8708664 TI - Neurofibrillary tangles in the brain of a 16 month old infant. PMID- 8708665 TI - Dysarthria-clumsy hand syndrome due to infarction of the cerebral peduncle. PMID- 8708666 TI - Transient alien hand syndrome: is this a seizure or a transient ischaemic attack? PMID- 8708667 TI - Brain water intoxication and CT hypolucencies in ovarian hyperstimulation syndrome complicating gametes intrafallopian transfer (GIFT) and tubal pregnancy. PMID- 8708668 TI - Thyrotoxic Hashimoto's encephalopathy. PMID- 8708669 TI - Cerebral salt wasting syndrome. PMID- 8708670 TI - A patient with cerebral autosomal dominant arteriopathy with subcortical infarcts and leucoencephalopathy (CADASIL) confirmed by sural nerve biopsy. PMID- 8708671 TI - Paraneoplastic necrotising myelopathy in a case of AIDS with lymphoma. PMID- 8708672 TI - Total tau protein immunoreactivity in lumbar cerebrospinal fluid of patients with Alzheimer's disease. PMID- 8708673 TI - Bilateral infarction of the rostral pontine tegmentum as a cause of isolated bilateral supranuclear sixth nerve palsy related to hypertension. PMID- 8708674 TI - Hemiballism in Parkinson's disease. PMID- 8708675 TI - Prospective evaluation of a prognostic index for intrinsic supratentorial tumours. PMID- 8708677 TI - Joseph Hyrtl (1811-94). PMID- 8708676 TI - The basis for behavioural disturbances in dementia. PMID- 8708679 TI - Small medullary infarction presenting as painful trigeminal sensory neuropathy. PMID- 8708678 TI - Neuroepidemiology of amyotrophic lateral sclerosis: clues to aetiology and pathogenesis. PMID- 8708680 TI - Creutzfeldt-Jakob disease in Austria. AB - Between 1969 and 30 September 1995, 79 Austrian patients had Creutzfeldt-Jakob disease (CJD) diagnosed neuropathologically by necropsy or biopsy. The annual incidence has significantly increased in recent years (average 0.18 per million in 1969-85, and 0.67 per million in 1986-94; estimate for 1995: 1.5 per million). Also, the percentage of patients with CJD over 70 years at death increased significantly until 1989 but is since in decline. There is no regional clustering, familial occurrence, or recognised iatrogenic risk. One patient had a 10 year history of intramuscular injection of purified bovine RNA preparation (Regeneresen) from various organs including the brian. The ages at death are symmetrically distributed around the median of 64 years. The median duration of disease is four months. Most patients (76%) died within six months of onset. Retrospectively, 86% of patients fulfilled clinical criteria of probable or possible CJD. Neuropathology showed the classic triad of spongiform change, astrogliosis, and neuronal loss in most cases. Two cases did not show unequivocal tissue alterations, but anti-PrP immunocytochemistry detected PrP deposits also in these cases. It is concluded that the recent rise in incidence of CJD in Austria most likely reflects increased awareness and diagnosis of CJD rather than a real increase. As bovine spongiform encephalopathy (BSE) has not been reported in Austria, the data do not support a link between a rise in incidence of sporadic CJD and BSE. PMID- 8708681 TI - Incidence of intracranial tumours in the Lothian region of Scotland, 1989-90. AB - OBJECTIVE: To determine the incidence of primary and secondary intracranial tumours in the Lothian region of south east Scotland. METHODS: A population based study was performed. Patients from Lothian with incident intracranial tumours diagnosed in 1989 and 1990 (by CT or histology) were identified retrospectively using multiple sources. Differences in incidence by tumour type, age, sex, and socioeconomic status were examined. RESULTS: Four hundred and forty two patients with incident intracranial tumours were identified (228 primary tumours and 214 secondary tumours). The crude yearly incidences of primary and secondary tumours were 15.3 and 14.3 per 100,000 respectively. The commonest primary tumours were neuroepithelial tumours (53.5%), meningeal tumours (19.5%), and sellar tumours (16.5%). About 50% of patients with secondary tumours had an underlying lung cancer. The incidence of primary and secondary tumours increased markedly with age. Meningeal tumours were more common in women, and neuroepithelial tumours were more common in those who lived in more affluent areas. CONCLUSIONS: The incidence rates of primary and secondary intracranial tumours in Lothian were more than twice those previously reported in the United Kingdom. Intracranial tumours are a significant cause of morbidity and mortality in the United Kingdom, and further research into their aetiology and treatment is urgently required. PMID- 8708682 TI - Startle provoked epileptic seizures:features in 19 patients. AB - OBJECTIVES: To define the clinical characteristics of a group of patients with startle provoked epileptic seizures (SPES). METHODS: Nineteen patients were identified during the course of a larger study of clinical seizure patterns. A witnessed seizure account was obtained in all patients; interictal EEG in 18, video-EEG-telemetry in eight, CT in 18, and high resolution MRI in eight. RESULTS: The onset of SPES was in childhood or adolescence in 14 of 19 patients. It was preceded by exclusively spontaneous seizures in nine patients and SPES had been replaced by exclusively spontaneous seizures in two patients. Sudden noise was the main triggering stimulus and somatosensory and visual stimuli were also effective in some patients. The clinical seizure pattern involved asymmetric tonic posturing in 16 of 19 patients. Focal neurological signs were present in nine patients, mental retardation in six, and 10 were clinically normal. Ictal scalp EEG showed a clear seizure discharge in only one patient with a tonic seizure pattern; over the lateral frontal electrodes contralateral to the posturing limbs. Brain CT showed a porencephalic cyst in three patients, focal frontal atrophy in one, and generalised atrophy in one. Brain MRI was undertaken in five normal subjects and three neurologically impaired patients, six with normal CT. It showed a porencephalic cyst in one patient. In six patients, there were dysplastic lesions. They affected the lateral premotor cortex in three patients and the perisylvian cortex in three patients, one with bilateral perisylvian abnormality. CONCLUSIONS: SPES are more frequent than is generally appreciated. They may be transient and occur relatively commonly without fixed deficit, by contrast with previous reports. The imaging abnormalities identified in those without diffuse cerebral damage suggest that SPES are often due to occult congenital lesions and that the lateral premotor and perisylvian cortices are important in this phenomenon. PMID- 8708683 TI - Brain atrophy in frontotemporal dementia. AB - OBJECTIVES: To evaluate the pattern of regional brain atrophy in patients with frontotemporal dementia by comparing it with that in patients with Alzheimer's disease and normal controls. METHODS: Fourteen patients with frontotemporal dementia, 13 with moderate, and 33 with mild Alzheimer's disease, and 31 controls were studied. Atrophy was evaluated with linear measures in the anterior brain, medial temporal lobe, and hippocampal formation regions using MRI. RESULTS: Patients with frontotemporal dementia had greater atrophy in the anterior brain regions than patients with Alzheimer's disease or controls. Atrophy of the hippocampal formation, which best discriminates Alzheimer's disease from controls, was present also in patients with frontotemporal dementia. By contrast, atrophy of the medial temporal lobe, which is also present in Alzheimer's disease, was absent in frontotemporal dementia. CONCLUSION: A pattern of atrophy in the frontal lobes and hippocampal formation with sparing of the medial temporal lobe might be distinctive of frontotemporal dementia. Hippocampal involvement might not be specific for Alzheimer's disease and specific patterns of atrophy might be distinctive of some forms of degenerative dementia. PMID- 8708684 TI - Dynamics of cerebral blood flow and metabolism in patients with cranioplasty as evaluated by 133Xe CT and 31P magnetic resonance spectroscopy. AB - OBJECTIVE: Prolonged improvement in neurological and mental disorders has been seen after only cranioplasty in patients initially treated with external decompression for high intracranial pressure. The objective was to evaluate, using 133Xe CT and 31P magnetic resonance spectroscopy (MRS), how restoring the bone itself can influence cerebral blood flow and cerebral energy metabolism after high intracranial pressure is attenuated. METHODS: Seven patients (45-65 years old) who had undergone external decompression to prevent uncontrollable intracranial hypertension after acute subarachnoid haemorrhage were evaluated. Cerebral blood flow and metabolic changes were evaluated before and after cranioplasty. RESULTS: The ratio of phosphocreatine to inorganic phosphate (PCr/Pi), which is a sensitive index of cerebral energy depletion, was calculated and beta-ATP was measured. The cerebral blood flow value in the thalamus was normalised, from 44 (SD 9) to 56 (SD 8) ml/100 g/min (P < 0.01) and the value in the hemisphere increased from 26 (SD 3) to 29 (SD 4) ml/100 g/min on the side with the bone defect. The PCr/Pi ratio improved greatly from 2.53 (SD 0.45) to 3.01 (SD 0.24) (P < 0.01). On the normal side, the values of cerebral blood flow and PCr/Pi increased significantly (P < 0.01) after cranioplasty, possibly due to transneural suppression. The pH of brain tissue was unchanged bilaterally after cranioplasty. CONCLUSION: Cranioplasty should be carried out as soon as oedema has disappeared, because a bone defect itself may decrease cerebral blood flow and disturb energy metabolism. PMID- 8708685 TI - Botulinum toxin in writer's cramp: objective response evaluation in 31 patients. AB - OBJECTIVES: To quantify the treatment effect of local botulinum toxin injections in writer's cramp a newly developed rating scale of writing performance and a computer assisted analysis of writing speed were used in 31 patients undergoing botulinum toxin therapy. METHODS: Baseline data of the writer's cramp rating scale (WCRS, see appendix) and computer based writing speed analysis were compared with those obtained at the time of subjective best response as recorded during follow up visits. RESULTS: The mean dose injected per session was 133.2 units Dysport divided between two forearm muscles. Of all 124 injection sessions during mean follow up of one year 76% produced a good improvement. The most common side effect was weakness (72% of the follow up visits). The WCRS scores as assessed by a blinded videotape review by four independent raters showed good reliability between raters and a significant improvement after treatment (P < 0.001). The speed of pen movements showed a significant (P < 0.05) increase after treatment at subjective best effect recordings and a significant correlation with WCRS subscores, documenting the validity of the scale. CONCLUSION: The present study is the first to show significant effects of botulinum toxin treatment in patients with writer's cramp on the basis of a quantifiable scale for writing performance which correlates significantly with writing speed measurements. The WCRS as employed in this study might therefore prove a useful rating instrument in other studies assessing severity and treatment response in patients with writer's cramp. PMID- 8708686 TI - Defective imitation of gestures in patients with damage in the left or right hemispheres. AB - OBJECTIVES: Defective imitation of meaningless gestures has repeatedly been demonstrated in patients with apraxia and has been interpreted as being due to a deficit of motor execution. There is, however, controversy as to whether some impairment of imitation also occurs in patients with right brain damage. The aim was to compare defective imitation in patients with left and right brain damage and to explore whether there are qualitative differences between them. METHODS: Imitation was examined in 80 patients with left brain damage (LBD) and aphasia, 40 patients with right brain damage (RBD), and 60 controls for three types of gestures:hand positions, finger configurations, and combined gestures which required a defined hand position as well as a defined configuration of the fingers. RESULTS: Regardless of whether imitation of hand positions and finger configurations were tested each on their own or together, they showed differential susceptibility to RBD and LBD. Whereas imitation of finger configurations was about equally impaired in RBD and LBD, defective imitation of hand positions occurred almost exclusively in patients with LBD, and whereas controls as well as patients with RBD committed less errors with hand positions than with finger configurations, the reverse was the case in patients with LBD. CONCLUSIONS: The pattern of results goes against a deficit of motor execution as being the cause of defective imitation in patients with LBD, as it is difficult to see why such a deficit should affect proximal movements necessary for reaching hand positions more than differential finger movements. An alternative explanation would be that in patients with LBD errors are due to defective mediation by knowledge about the human body whereas in patients with RBD they stem from faulty visuospatial analysis of the demonstrated gesture. PMID- 8708687 TI - Nerve conduction studies in adrenomyeloneuropathy. AB - OBJECTIVE: Adrenomyeloneuropathy (AMN) is an X linked metabolic disorder presenting with progressive spastic paraparesis in the third to fifth decade of life. Although peripheral neuropathy is also present in most patients, prominent pyramidal signs may make its clinical recognition difficult. The objective was to characterise the peripheral neuropathy in patients with AMN by nerve conduction studies. METHODS: Nerve conduction studies were performed in 99 men known to have AMN and in 38 heterozygous women, all of whom had neurological disabilities. RESULTS: Of the 13 variables obtained, at least one was abnormal in 82% of patients. The abnormalities were more common in men than in women (87% v 67%); in legs than in arms (77% v 38%); in motor than in sensory conduction (80% v 39%); and in latency (distal and F wave) and velocity compared with amplitude (80% v 29%). Twenty six patients had at least one nerve variable value in the demyelinating range. Four variables (sural velocity, peroneal amplitude, peroneal velocity, and peroneal F wave) were correlated with the expanded disability status scale; five variables (peroneal velocity, tibial H reflex, median distal latency, median conduction velocity, and median F wave latency) were correlated with serum very long chain fatty acids (VLCFAs); and two variables (sural amplitude and peroneal distal latency) were more likely to be abnormal in patients with normal adrenal function than in patients with Addison's disease. CONCLUSIONS: Nerve conduction studies in patients with AMN are often abnormal and suggest a mixture of axonal loss and multifocal demyelination. Their correlation with disability status and serum VLCFAs suggests that measures from nerve conduction studies may be useful in evaluating future treatments. PMID- 8708689 TI - A case report of paroxysmal dystonic choreoathetosis due to hypoglycaemia induced by an insulinoma. AB - Hypoglycaemia due to an insulinoma can mimic acute disorders of cognition, consciousness, epilepsy, transient ischaemia, or psychosis, and chronic disorders of dementia and neuropathy. Misdiagnosis and delay in treatment are common and prolonged hypoglycaemia can lead to permanent neurological deficit or fatal coma. A 27 year old woman with hypoglycaemia induced by an insulinoma presented with features typical of paroxysmal non-kinesiogenic dystonic choreoathetosis. Striatal dysfunction as a consequence of hypoglycaemia has rarely been described. Insulinoma is a readily treatable condition that should be considered in the differential diagnosis of a paroxysmal movement disorder. PMID- 8708688 TI - Memory without context: amnesia with confabulations after infarction of the right capsular genu. AB - OBJECTIVE: To explore the mechanism of an amnesia marked by confabulations and lack of insight in a patient with an infarct of the right inferior capsular genu. The confabulations could mostly be traced back to earlier events, indicating that the memory disorder ensued from an inability to store the temporal and spatial context of information acquisition rather than a failure to store new information. METHODS: To test the patient's ability to store the context of information acquisition, two experiments were composed in which she was asked to decide when or where she had learned the words from two word lists presented at different points in time or in different rooms. To test her ability to store new information, two continuous recognition tests with novel non-words and nonsense designs were used. Recognition of these stimuli was assumed to be independent of the context of acquisition because the patient could not have an a priori sense of familiarity with them. RESULTS: The patient performed at chance in the experiments probing knowledge of the context of information acquisition, although she recognised the presented words almost as well as the controls. By contrast, her performance was normal in the recognition tests with non-words and nonsense designs. CONCLUSION: These findings indicate that the patient's amnesia was based on an inability to store the context of information acquisition rather than the information itself. Based on an analysis of her lesion, which disconnected the thalamus from the orbitofrontal cortex and the amygdala, and considering the similarities between her disorder, Wernicke-Korsakoff syndrome, and the amnesia after orbitofrontal lesions, it is proposed that contextual amnesia results from interruption of the loop connecting the amygdala, the dorsomedial nucleus, and the orbitofrontal cortex. PMID- 8708690 TI - Familial bulbospinal neuronopathy with optic atrophy: a distinct entity. AB - A 61 year old woman and her 58 year old brother presented with the clinical picture of late onset progressive bulbar and spinal muscular atrophy with family history of involvement in successive generations. The sister also had optic neuropathy and the brother developed diabetes mellitus and sex hormone abnormalities. Neurophysiological and histopathological studies showed a pattern of motor and sensory neuronopathy. There was no abnormal expansion of CAG repeats in the androgen receptor gene. This family seems to have a previously unrecognised entity with the bulbospinal neuronopathy phenotype. PMID- 8708691 TI - Disorders of gait. PMID- 8708692 TI - A special form of cerebral lacunae: expanding lacunae. AB - The case of a 42 year old man with headache, blurred vision, and diplopia allowed the description of a particular form of cerebral lacunae-that is, expanding lacunae. Brain MRI showed hydrocephalus and multiple lesions in the thalamomesencephalic region. The radiological features of these lesions were similar to the histological brain coronal section of a case reported in 1983 in which expanding lacunae were related to a dilatation of the perivascular spaces and a focal segmental necrotising angiitis. The role of the lymphatic drainage of the brain is discussed to explain the dilatation of the perivascular spaces. The hypothesis of a hydrodynamic factor being responsible for the expanding character of the lacunae was suggested by the location of the lesions and the influence of various clinical events on the symptomatology. PMID- 8708693 TI - Obstructive hydrocephalus due to benign cysts of the thalamus: report of two patients. PMID- 8708694 TI - Antineutrophil cytoplasmic antibodies and the optic-spinal form of multiple sclerosis in Japan. PMID- 8708696 TI - Isolated dysarthria caused by a right paravermal infarction. PMID- 8708695 TI - Cerebral venous sinus thrombosis associated with factor V gene mutation. PMID- 8708697 TI - Bilateral internal carotid artery agenesis. PMID- 8708698 TI - Possible distinct pathogenesis in low responder myasthenia gravis: association of soluble interleukin-2 receptor with acetylcholine receptor antibody titre or abnormal thymus. PMID- 8708699 TI - Decreased magnetisation transfer ratio due to demyelination: a case of central pontine myelinolysis. PMID- 8708700 TI - Raised plasma polyamine concentrations in patients with severe head injury. PMID- 8708701 TI - Hatred of the hemiparetic limbs (misoplegia) in a 10 year old child. PMID- 8708703 TI - Increased serum neopterin concentrations in a patient with Creutzfeldt-Jakob disease. PMID- 8708702 TI - Valproate induced encephalopathy treated with carnitine in an adult. PMID- 8708704 TI - Thalamic stimulation for severe action tremor after lesion of the superior cerebellar peduncle. PMID- 8708705 TI - Structural and functional changes in patients with motor neuron disease. PMID- 8708706 TI - Should we dream the impossible dream? The meaning of long-term survival in metastatic breast cancer. PMID- 8708707 TI - Autologous stem-cell transplantation and purging. PMID- 8708708 TI - Long-term follow-up of patients with complete remission following combination chemotherapy for metastatic breast cancer. AB - PURPOSE: To determine the long-term clinical course of patients with metastatic breast cancer (MBC) who achieved a complete remission with doxorubicin-alkylating agent-containing combination chemotherapy programs. PATIENTS AND METHODS: To assess the long-term prognosis of MBC, we reviewed our experience with 1,581 patients treated on consecutive doxorubicin and alkylating agent-containing front line treatment protocols between 1973 and 1982. Treatment was administered for a maximum duration of 2 years. Characteristics of long-term survivors were evaluated, and hazard rates for progression were calculated. RESULTS: From this group, 263 (16.6%) achieved complete responses (CR) and 49 (3.1%) remained in CR for more than 5 years. After a median duration of 191 months, 26 patients remain in first CR, four patients died in CR at times ranging from 118 to 234 months, 18 patients died of breast cancer, and one is alive with metastatic disease. Compared with the overall CR and total patient populations, the long-term CR group had more premenopausal patients, a younger median age, a lower tumor burden, and better performance status. The hazard function shows a substantial drop in risk of progression after approximately 3 years from initiation of therapy. Ten long-term CR patients developed second primary cancers: breast (3), ovary (2), pancreas (1), endometrium (1), colon (1), head and neck (1), and lung (1). CONCLUSION: Most patients with MBC treated with systemic therapies have only temporary responses to treatment, but some patients continue in CR following initial treatment. These data show that a small percentage of patients achieve long-term remissions with standard chemotherapy regimens. Remission consolidation strategies are needed. PMID- 8708710 TI - Age and serum lactate dehydrogenase level are independent prognostic factors in human immunodeficiency virus-related non-Hodgkin's lymphomas: a single-institute study of 96 patients. AB - PURPOSE: The role of classical pragnostic factors (ie, age, performance status [PS], stage, extranodal involvement, and serum lactate dehydrogenase [LDH] level) included in the International Index for diffuse large-cell non-Hodgkin's lymphoma (NHL) of the general population is presently unknown in the setting of human immunodeficiency virus (HIV). To assess the prognostic value of these factors in HIV-related NHL, we reviewed the cohort of patients with HIV-related NHL diagnosed and treated with combination chemotherapy (CT) at our institution. PATIENTS AND METHODS: Ninety-six patients with systemic HIV-related NHL diagnosed and treated with combination CT regimens between September 1987 and December 1993 at the Centro di Riferimento Oncologico, Aviano, Italy, were studied. All clinical and laboratory data were evaluated by univariate and multivariate analyses, using overall survival as the end point. RESULTS: Complete remission (CR) occurred in 48% of patients; the overall median survival and disease-free survival times were 7 and 13 months, respectively. Among the classical and HIV related prognostic factors, the following had a statistically significant influence on survival: PS > or = 2, elevated LDH level, age greater than 40 years, a CD4 cell count less than 100/microL, active opportunistic infections at diagnosis of NHL, and B symptoms. Multivariate analyses showed that only age, serum LDH level, and CD4 cell count were independent predictors of shortened survival. The increased hazard for patients greater than 40 years of age was 1.6 (95% confidence interval [CI], 1.2 to 2.3), for patients with increased LDH it was 1.8 (95% CI, 1.01 to 3.1), and for patients with a CD4 cell count less than 100/microL it was 1.7 (95% CI, 1.01 to 2.9). CONCLUSIONS: Our study shows that in addition to HIV-related prognostic factors, ie, CD4 cell count less than 100/microL, classical prognostic factors such as age and serum LDH level are independent prognostic factors and should be included in the design of future clinical trials of HIV-related NHL. PMID- 8708709 TI - In vivo purging with high-dose cytarabine followed by high-dose chemoradiotherapy and reinfusion of unpurged bone marrow for adult acute myelogenous leukemia in first complete remission. AB - PURPOSE: To evaluate in a prospective study the efficacy of autologous bone marrow transplantation (BMT) in adult patients with acute myelogenous leukemia (AML) in first remission, using a single course of high-dose Cytarabine (HD Ara C) consolidation therapy as in vivo purging. PATIENTS AND METHODS: Sixty consecutive adult patients with AML in first complete remission (CR) were treated with HD Ara-C consolidation therapy as a method of in vivo purging before marrow collection. High-dose therapy consisted of fractionated total-body irradiation (FTBI) 12 Gy, intravenous etoposide 60 mg/kg, and cyclophosphamide 75 mg/kg, followed by reinfusion of cryopreserved marrow. RESULTS: Sixty patients underwent consolidation treatment with HD Ara-C with the intent to treat with autologous BMT. Sixteen patients were unable to proceed to autologous BMT (10 patients relapsed, one died of sepsis, one developed cerebellar toxicity, two had inadequate blood counts, and two refused). Forty-four patients underwent autologous BMT and have a median follow-up time of 37 months (range, 14.7 to 68.7) for patients who are alive with no relapse. The cumulative probability of disease-free survival (DFS) at 24 months in the intent-to-treat group is 49% (95% confidence interval [CI], 37% to 62%) and in those who actually underwent autologous BMT is 61% (95% CI, 46% to 74%). The probability of relapse was 44% (95% CI, 31% to 58%) and 33% (95% CI, 20% to 49%) for the intent-to-treat and autologous BMT patients, respectively. CONCLUSION: This approach offers a relatively high DFS rate to adult patients with AML in first CR. The results of this study are similar to those achieved with allogeneic BMT. PMID- 8708711 TI - Highly purified CD34-positive cells reconstitute hematopoiesis. AB - PURPOSE: The objective of this study was to characterize CD34+ cell grafts, obtained using a novel technique, from children undergoing autologous bone marrow transplantation (BMT) for cancer therapy. In particular, we wanted to determine if the CD34+ marrow cell grafts generated hematopoietic reconstitution, since a positive result would motivate further development and use of this methodology. PATIENTS AND METHODS: This pilot feasibility clinical trial involved 13 patients < or = 25 years of age with advanced solid tumors, including seven children with neuroblastoma. Harvested bone marrow underwent immunomagnetic CD34+ selection. RESULTS: In three of 13 enrolled patients, low purities of the CD34+ preparations disqualified the use of the CD34+ marrow grafts. Ten patients received myeloablative chemotherapy with etoposide, carboplatin, and cyclophosphamide, then were transplanted with CD34+ marrow grafts. In the 10 patients transplanted with CD34(+)-selected cells, the CD34+ cell purity (nucleated RBCs excluded) in the cell graft preparation was 91% total cell recovery from the starting light density cells 2.2%, CD34+ cell recovery 38%, colony-forming unit-granulocyte macrophage (CFU-GM) recovery 23%, and estimated tumor-cell depletion 2.6 logs (medians). The CD34+ marrow grafts administered to these patients contained a median of 2.3 x 10(6) nucleated cells, 1.4 x 10(6) CD34+ cells, and 1.3 x 10(4) CFU-GM per kilogram patient weight. Most patients experienced only the toxicities previously observed with this myeloblative chemotherapy regimen, although two unusual toxicities were observed. All 10 patients transplanted with CD34+ cell grafts engrafted. CONCLUSION: The CD34+ purified grafts were enriched in stem/progenitor cells, with five of these 10 preparations containing > or = 94% CD34+ cells. Engraftment with CD34(+)-purified cell grafts as pure as 99% confirms that autologous CD34+ cells, alone, are sufficient to provide hematopoietic rescue for myeloablated patients. The best purification results were obtained on small marrow harvests from patients with neuroblastoma. The engraftment of highly purified CD34+ cells obtained by this technology and the antitumor effect of the transplant, by which two of 10 poor prognosis patients remain clinically free of tumor, have stimulated further clinical trials. PMID- 8708712 TI - Phase I study of subcutaneously administered interleukin-2 in combination with interferon alfa-2a in patients with advanced cancer. AB - PURPOSE: Although high-dose interleukin-2 (IL-2) can produce durable remissions in a subset of responding patients with renal cell carcinoma (RCC), this occurs in the setting of significant toxicity. The purpose of this study is to define the maximum-tolerated dosage (MTD) of IL-2 and interferon alfa-2a (IFN alpha-2a) that can be administered chronically on an outpatient basis. PATIENTS AND METHODS: Fifty-three patients with advanced cancer of variable histology with good prognostic features were treated in six cohorts. Patients in cohorts one through five received IL-2 (1.5 or 3.0 x 10(6) million units (mU)/m2) Monday through Friday and IFN alpha-2a (1.5 or 3 x 10(6) mU/m2) daily for a 4-week cycle. In cohort six, IFN alpha-2a was given three times a week. Immunologic monitoring, including serum levels of soluble IL-2 receptor (sIL-2R) and neopterin, flow cytometry, and natural killer cell (NK) activity, were measured. Patients were evaluated for toxicity, response, and survival. RESULTS: Almost all patients developed grade I/II toxicities commonly associated with cytokine therapy. Symptoms were most severe with the first treatment of each week. Dose limiting toxicities included grade III fatigue, hypotension, and creatinine elevations. The MTD was 1.5 mU/m2 daily x 5 given subcutaneously repeated weekly for IL-2 and 1.5 mU/m2 daily subcutaneously (dose level 3) for IFN. Six of 25 assessable patients with RCC (24%) achieved a partial response (PR), including four of eight patients who were previously untreated. There were no objective responses in patients with other tumors, including 12 melanoma patients. CONCLUSION: IL-2 and IFN alpha-2a can be given with tolerable toxicities on an outpatient basis and shows significant activity in patients with metastatic RCC. PMID- 8708713 TI - Double-blind, randomized comparison of the antiemetic efficacy of intravenous dolasetron mesylate and intravenous ondansetron in the prevention of acute cisplatin-induced emesis in patients with cancer. Dolasetron Comparative Chemotherapy-induced Emesis Prevention Group. AB - PURPOSE: To assess the comparative antiemetic efficacy of single-dose intravenous (IV) dolasetron mesylate and ondansetron in preventing cisplatin-induced nausea and vomiting. PATIENTS AND METHODS: Cancer patients (n = 609) receiving first course cisplatin chemotherapy were randomized to one of three treatments: 1.8 or 2.4 mg/kg dolasetron mesylate salt (equivalent to 1.3 and 1.8 mg/kg dolasetron base, respectively) or 32 mg ondansetron. Each treatment was infused over 15 minutes, 30 minutes before cisplatin administration. Patients were stratified to cisplatin doses of > or = 70 and less than 91 mg/m2 (n = 368) or > or = 91 mg/m2 (n = 241), administered over < or = 3 hours. Protocol-defined efficacy criteria included complete response (zero emetic episodes and no rescue medication), major response (1 to 2 emetic episodes and no rescue medication), and patients' report of nausea severity and satisfaction recorded on a 100-mm visual analog scale (VAS). RESULTS: The three treatments met protocol-specified criteria for equivalence. Complete response rates for dolasetron mesylate 1.8 mg/kg, 2.4 mg/kg, and ondansetron, respectively, were 49.2%, 45.6%, and 50.4% for patients in the lower cisplatin stratum (mean, 74.7 mg/m2) and 36.8%, 31.3%, and 31.8% in the higher cisplatin stratum (mean, 100.6 mg/m2). No significant differences were observed in the extent of nausea with either dolasetron dose compared with ondansetron. Less nausea was noted with 1.8 mg/kg dolasetron compared with the 2.4 mg/kg dose (P = .044) All three antiemetic treatments were well tolerated. Asymptomatic electrocardiogram changes were recorded with both dolasetron and ondansetron. CONCLUSION: A single IV dose of dolasetron mesylate (1.8 or 2.4 mg/kg) has comparable safety and efficacy to a single 32-mg IV dose of ondansetron in patients receiving cisplatin chemotherapy. PMID- 8708714 TI - Double-blind, randomized study of primary hormonal treatment of stage D2 prostate carcinoma: flutamide versus diethylstilbestrol. AB - PURPOSE: Patients with stage D2 prostate carcinoma are often treated initially with hormones to decrease endogenous testosterone. Therapy with diethylstilbestrol (DES), leuprolide, or bilateral orchiectomy has been reported to be equivalent. DES is the cheapest preparation, but has the potential for serious cardiovascular or thromboembolic complications. Flutamide is a novel antiandrogen with fewer side effects. PATIENTS AND METHODS: The Eastern Cooperative Oncology Group (ECOG) conducted a double-blind, randomized study to compare the efficacy of flutamide (250 mg three times daily) to DES (1 mg three times daily) as the primary hormonal therapy for patients with stage D2 prostate cancer. Patients were stratified by performance status, disease sites, and history of cardiovascular disease at randomization. RESULTS: Forty-eight patients received DES and 44 flutamide. Patient characteristics were evenly distributed between the two treatments. The overall response rate was similar (DES, 62%; flutamide, 50%). Grade III or worse cardiovascular or thromboembolic toxicity developed in 33.3% of patients on DES and in 17.6% on flutamide (P = .051). Other toxicities were similar between the two treatment arms. However, DES produced significantly longer time to treatment failure (26.4 v 9.7 months, P = .016) and longer survival than flutamide (43.2 v 28.5 months, P = .040). CONCLUSION: As the primary hormonal therapy for stage D2 prostate cancer, DES caused more serious cardiovascular or thromboembolic complications than flutamide. Despite this, flutamide was not as active an initial agent as DES. However, the effectiveness of flutamide in conjunction with other agents compared with DES remains undetermined, and the optimal initial hormone therapy of stage D2 prostate cancer requires further studies. PMID- 8708715 TI - Outcomes of external-beam radiation therapy for prostate cancer: a study of Medicare beneficiaries in three surveillance, epidemiology, and end results areas. AB - PURPOSE: This study was designed to obtain representative estimates of the quality of life and probabilities of possible adverse effects among Medicare-age patients treated with external-beam radiation therapy for prostate cancer. METHODS: Patients treated for local or regional prostate cancer with high-energy external-beam radiation between 1989 and 1991 were sampled from a claims data base of the Surveillance, Epidemiology, and End Results (SEER) program from three regions. Patients were surveyed primarily by mail, with telephone follow-up evaluation of non-respondents. There were 621 respondents (83% response rate). The results were compared with data from a previously published national survey of Medicare-age men who had undergone radical prostatectomy. RESULTS: Although they were older at the time of treatment, radiation patients were less likely than surgical patients to wear pads for wetness (7% v 32%) and had a lower rate of impotence (23% v 56% for men < 70 years), while they were more likely to report problems with bowel dysfunction (10% v 4%). Both groups reported generally positive feelings about their treatments. Radiation and surgical patients reported similar rates of additional subsequent treatment (24% v 26% at 3 years after primary treatment). However, radiation patients were less likely to say they were cancer-free, and they reported more worry about cancer than did surgical patients. CONCLUSION: The health-related quality of life of radiation and surgical patients, on average, is similar, but the pattern of experience with adverse consequences of treatment differs by treatment. PMID- 8708716 TI - Modulation of high-dose infusional fluorouracil by low-dose methotrexate in patients with advanced or metastatic colorectal cancer: final results of a randomized European Organization for Research and Treatment of Cancer Study. AB - PURPOSE: Methotrexate (MTX) has been described to modulate the activity of fluorouracil (5-FU) in patients with metastatic colorectal cancer. The European Organization for Research and Treatment of Cancer (EORTC) Gastrointestinal Tract Cancer Cooperative Group (GITCCG) conducted a phase III trial to investigate the efficacy and tolarability of the addition of low-dose MTX (40 mg/m2) to high-dose infusional 5-FU (60 mg/kg over 48 hours) given weekly for 4 weeks and thereafter every 2 (for 4 weeks) and 3 weeks. PATIENTS AND METHODS: Three hundred ten patients were randomized between 1987 and 1992. Eligible patients had measurable advanced or metastatic colorectal cancer and had not been pretreated with antifolates or fluorodinated pyrimidines. All 297 eligible patients were evaluated for survival; toxicity was assessed in 292 patients who received at least one course of treatment. Patients with bidimensionally measurable disease (n = 230) were also evaluated for response according to standard criteria. RESULTS: The addition of low-dose MTX to high-dose infusional 5-FU led to a doubling of the response rate from 10% to 21% (P = .025). The median survival time also increased from 9.3 to 12.5 months, but this difference was not statistically significant (P = .12). High-dose infusional 5-FU with or without low-dose MTX was well tolerated, with grade 3 to 4 toxicity in greater than 10% of patients only occurring for stomatitis with the combination treatment. Performance status was the sole prognostic factor for survival in a multivariate analysis. CONCLUSION: Low-dose MTX effectively modulated high-dose infusional 5 FU in a large, randomized trial in which less than 5% of patients received leucovorin. PMID- 8708717 TI - Weekly high-dose leucovorin versus low-dose leucovorin combined with fluorouracil in advanced colorectal cancer: results of a randomized multicenter trial. Study Group for Palliative Treatment of Metastatic Colorectal Cancer Study Protocol 1. AB - PURPOSE: To determine the most effective dose of leucovorin (folinic acid [FA]) within a weekly bolus fluorouracil (FU) schedule, we conducted a randomized multicenter trial to compare therapeutic effects and toxicity of high-dose FA versus low-dose FA combined with FU at equal doses in both treatment groups. PATIENTS AND METHODS: Patients with measurable inoperable or metastatic colorectal cancer were randomized to receive weekly FU 500 mg/m2 by intravenous (IV) bolus combined with high-dose FA 500 mg/m2 (group A) or low-dose FA 20 mg/m2 (group B) by 2-hour infusion. RESULTS: Of 291 assessable patients (group A, n = 148; group B, n = 143), we observed, in group A, complete response (CR)/partial response (PR) in 32 (21.6%), no change (NC) in 64 (43.2%), and progressive disease (PD) in 52 (35.1%); and in group B, CR/PR in 25 (17.5%), NC in 63 (44.1%), and PD in 55 (38.5%). The median response duration was 24.8 weeks in group A and 23.1 weeks in group B. Median progression-free intervals were 29.3 weeks (group A) and 30 weeks (group B). The median survival time was 55.1 weeks in group A and 54.1 weeks in group B. Overall toxicity was moderate. Mild nausea and vomiting, and stomatitis were common side effects in both groups. The incidence of World Health Organization (WHO) grade III/IV diarrhea was significantly higher in group A (40 v 23 patients). Severe side effects were observed only in a minority of patients in both arms. WHO grade IV diarrhea was observed in seven patients: four in group A and three in group B. The rate of toxicity-related adjustments of dose and schedule was comparable in both groups. CONCLUSION: High-dose FA/FU is not superior to low-dose FA/FU within a weekly treatment schedule. Response rates and survival were comparable in both treatment arms. Treatment-related toxicity was higher in group A (high-dose FA). Therefore, low-dose FA combined with weekly FU may be considered the preferred treatment for metastatic colorectal cancer. PMID- 8708718 TI - Randomized trial assessing the addition of interferon alpha-2a to fluorouracil and leucovorin in advanced colorectal cancer. Colorectal Cancer Working Party of the United Kingdom Medical Research Council. AB - PURPOSE: To determine the effects of interferon alpha-2a (IFN alpha) on the efficacy and toxicity of fluorouracil (FUra) and leucovorin (LV) in patients with advanced colorectal cancer. PATIENTS AND METHODS: Two hundred sixty chemotherapy naive patients were randomized to FUra/LV alone or FUra/LV plus IFN alpha. All patients received: LV 200 mg/m2 intravenous (IV) infusion over 2 hours, then FUra 400 mg/m2 i.v. bolus plus 400 mg/m2 i.v. infusion over 22 hours, all repeated on day 2. Treatment was every 2 weeks for up to 12 cycles. Patients randomized to IFN alpha received 6 x 10(6) IU subcutaneously every 48 hours throughout. Objective response (OR) and toxicity were assessed conventionally; in addition, palliative benefit and adverse effects were assessed using quality-of-life (QoL) questionnaires. RESULTS: There were no differences in OR rate, progression-free survival, or overall survival. OR rates in assessable patients were as follows: FUra/LV alone (n = 104), complete or partial response (OR) = 27%, no change (NC) = 34%; FUra/LV/IFN alpha (n = 101), OR = 28%, NC = 30%. Median survival was 10 months in both arms. Dose-limiting FUra toxicities were not significantly increased by co-administration of IFN alpha, and the delivered FUra dose intensity was not significantly reduced. However, QoL was adversely affected: patients on IFN alpha were less likely to report improvement in pretreatment physical and psychologic symptoms, and more likely to report new or worsening symptoms. CONCLUSION: IFN alpha, at a dose that impaired QoL, did not improve the efficacy of FUra/LV. The power of this trial is sufficient to exclude with 95% confidence a benefit of 15% in OR or 10 weeks in median survival. Accordingly, we cannot recommend the use of IFN alpha as a clinical modulator of FUra/LV in the treatment of advanced colorectal cancer. PMID- 8708719 TI - Lymph node retrieval in a randomized trial on western-type versus Japanese-type surgery in gastric cancer. AB - PURPOSE: In the tumor-node-metastasis (TNM) staging system, no recommendations are provided on what lymph node retrieval technique is to be used to determine lymph node status, which leads to variability in nodal status assessment and TNM staging. PATIENT AND METHODS: Lymph node retrieval was quantitated using data from 237 curatively resected gastric cancer patients, from a prospective, randomized trial that compared the Western resection with limited (D1) and the Japanese resection with extended lymphadenectomy (D2), and compared data from the literature. Moreover, the efficacy of different lymph node retrieval techniques was determined. RESULTS: The mean yield of lymph nodes was 15 in D1 and 30 in D2, which is similar to results from German investigators, but substantially lower than results from Japanese investigators (60 in D2). Use of a fat-clearance technique significantly increased (P = .01) nodal yields compared with conventional retrieval. Significantly higher yields (P < .001) were obtained by a Japanese surgeon using conventional retrieval directly postoperatively. Experience of surgicopathologic teams with processing resection specimens did not influence nodal yields. Further analysis showed that reference values for nodal yields per anatomically defined station as reported in the literature were contradicted by our results and indicated the ambiguity of such standards. CONCLUSION: Despite some anatomical variability in the distribution of lymph nodes, advice on the number of nodes to examine per N level, feasible in all patients, should be incorporated into the TNM classification to standardize nodal status assessment. Based on our findings, we advocate retrieval of nodes immediately postoperatively by the surgeon. PMID- 8708720 TI - Clinical utility of external immunoscintigraphy with the IMMU-4 technetium-99m Fab' antibody fragment in patients undergoing surgery for carcinoma of the colon and rectum: results of a pivotal, phase III trial. The Immunomedics Study Group. AB - PURPOSE: To assess the performance and the potential clinical impact of a new antibody imaging agent, CEA-Scan (Immunomedics Inc, Morris Plains, NJ), in 210 presurgical patients with advanced recurrent or metastatic colorectal carcinomas. METHODS: CEA-Scan, an anti-carcinoembryonic antigen (CEA) Fab antibody fragment labeled with technetium-99m-pertechnetate (99mTc), was injected intravenously (IV), and external scintigraphy was performed 2 to 5 and 18 to 24 hours later. Imaging with conventional diagnostic modalities (CDM) was also performed, and findings were confirmed by surgery and histology. RESULTS: The sensitivity of CEA Scan was superior to that of CDM in the extrahepatic abdomen (55% v 32%; P = .007) and pelvis (69% v 48%; P = .005), and CEA-Scan findings complemented those of CDM in the liver. Among 122 patients with known disease, the positive predictive value was significantly higher when both modalities were positive (98%) compared with each alone (68% to 70%), potentially obviating the need for histologic confirmation when both tests are positive. Imaging accuracy also was significantly improved by adding CEA-Scan to CDM. In 88 patients with occult cancer, imaging accuracy was enhanced significantly by CEA-Scan combined with CDM (61% v 33%). Potential clinical benefit from CEA-Scan was demonstrated in 89 of 210 patients. Only two patients developed human antimouse antibodies (HAMA) to CEA-Scan after a single injection, and none of 19 assessable patients after two injections. CONCLUSION: CEA-Scan affords high-quality, same-day imaging, uses an inexpensive and readily available radio-nuclide, adds clinically significant information in assessing extent and location of disease in colorectal cancer patients, and only rarely induces a HAMA response. PMID- 8708721 TI - Phase II study of paclitaxel therapy for unresectable biliary tree carcinomas. AB - PURPOSE: To evaluate the efficacy of paclitaxel administered to patients with unresectable adenocarcinomas of the gallbladder and biliary tree over 3 hours every 21 days. PATIENTS AND METHODS: Fifteen patients with unresectable and/or metastatic carcinoma of the gallbladder and bile ducts received intravenous paclitaxel over 3 hours after premedication with dexamethasone, diphenhydramine, and cimetidine. Treatment was repeated every 21 days, and one complete course of therapy was comprised of two such 21-day treatment cycles. The initial dose of paclitaxel was 170 mg/m2, and this was elevated to 200 mg/m2 due to tolerance within the initial patient cohort. RESULTS: All patients were assessable for both toxicity and response: 11 with bile duct cancer and four with gall-bladder carcinoma. Forty-three cycles of therapy were delivered during the trial (median, two), and one patient remains on treatment. No complete or partial responses were noted, although two patients achieved minor responses that lasted 2 and 2+ months, respectively. There were no deaths on this study, and all but one of the patients is still alive. The therapy was well tolerated, and hematologic and mucosal toxic effects were moderate and readily reversible, although significant neuromuscular adverse effects were noted. CONCLUSION: These findings indicate that paclitaxel, administered on this schedule, is tolerable, but is unlikely to have activity in metastatic carcinomas of the biliary tree. It is unclear whether a different regimen of paclitaxel, or another taxane, may have activity in these neoplasms. PMID- 8708722 TI - Phase II trial of intravenous flourouracil and subcutaneous interferon alfa-2b for biliary tract cancer. AB - PURPOSE: To assess the efficacy of systemic intravenous-fluorouracil (5-FU) and subcutaneous recombinant human interferon alfa-2b (rIFN alpha-2b) in patients with measurable cancer of the biliary tree. PATIENTS AND METHODS: Thirty-five patients (25 with cholangiocarcinoma and 10 with gallbladder carcinoma) were registered onto this phase II protocol between 1992 and 1995. Patients received a continuous infusion of 750 mg/m2/d of 5-FU on days 1 through 5 through a centrally placed venous catheter and a subcutaneous injection of 5 MU/m2 of rIFN alpha-2b on days 1, 3, and 5. Treatment cycles were repeated every 14 days; one course of therapy included four treatment cycles. Disease status was assessed every 8 weeks. Dosages were lowered for grade III mucositis. Fourteen patients had prior treatment and, before initiating this therapy, 17 patients required decompression of the biliary tree. RESULTS: Eleven of 32 (34%) assessable patients had a partial response. The median time to disease progression was 9.5 months, and the median survival time 12 months. Grade III to IV toxic effects were granulocytopenia (14%), mucositis (20%), diarrhea (9%), and dermatitis (11%). Grade III to IV asthenia and fatigue were observed in 6% of patients. CONCLUSION: Drug tolerance was better among previously untreated patients. To achieve a complete response, additional chemotherapy or radiotherapy should be considered when liver resection or transplantation is not feasible. However, if these results can be reproduced by other investigators, the regimen should be studied for adjuvant treatment of gallbladder carcinoma incidentally identified in patients undergoing cholecystectomy. PMID- 8708723 TI - Phase II study of paclitaxel in patients with recurrent malignant glioma. AB - PURPOSE: To assess the efficacy and toxicity of paclitaxel administered as a 3 hour infusion to patients with recurrent malignant glioma. PATIENTS AND METHODS: Adult patients with recurrent malignant glioma following radiation therapy, who had received no more than one prior chemotherapy regimen and who had a Karnofsky performance status (KPS) > or = 60, were treated with a 3-hour infusion of paclitaxel every 3 weeks. The initial dose was 210 mg/m2; dose escalation to 240 mg/m2 was allowed. Tumor response was assessed at 6-week intervals using radiographic and clinical criteria. Treatment was continued until documented tumor progression or a total of 12 paclitaxel infusions. RESULTS: Of 41 eligible patients, all were assessable for treatment toxicity and 40 (98%) were assessable for response. The response rate (disease stabilization or better) was 35%. Twenty nine patients (71%) underwent dose escalation to 240 mg/m2 without the use of growth factors. Toxicities included alopecia (98%), nausea (22%), arthralgias (32%), CNS toxicity (24%), peripheral neuropathy (15%), cardiac toxicity (7%), and myelosuppression (10% grade 3 or 4 hematologic toxicity). No patient developed febrile neutropenia. There was one allergic reaction (2%). CONCLUSION: Paclitaxel is well tolerated at this dose schedule in patients with recurrent malignant glioma, and affords a modest response rate. Because minimal myelotoxicity was encountered in our patients, a dose-escalating phase I/II study of paclitaxel is planned to determine the maximal-tolerated dose (MTD). PMID- 8708724 TI - Chemotherapy followed by accelerated fractionated radiation for larynx preservation in patients with advanced laryngeal cancer. AB - PURPOSE: Larynx preservation in advanced, resectable laryngeal cancer may be achieved using induction chemotherapy (CT) followed in responding patients by definitive radiation (RT). To address potential accelerated repopulation of clonogenic tumor cells during the prolonged total treatment time, we studied the feasibility of accelerated fractionated RT after CT. METHODS: Patients with advanced laryngeal cancer received two cycles of cisplatin 100 mg/m2 and fluorouracil (5-Fu) 1,000 mg/m2/d for 5 days. Responding patients received a third cycle after which those who had complete response or tumor down-staging to T1 proceeded with accelerated RT: 70.4 Gy delivered over 5.5 weeks. Patients who achieved a lesser response to CT underwent total laryngectomy and postoperative RT. RESULTS: Thirty-three patients were accrued. Three died during the course of CT and two declined definitive treatment after CT. Twenty-one patients had a major response to CT, 20 of whom received accelerated RT. Median weight loss during RT was 11%. Late severe morbidity was observed in five patients (25%). All four patients who underwent salvage laryngectomy after accelerated RT experienced major postoperative complications. The locoregional failure rate was 25%. The larynx was preserved in 48% of the total study population and in 80% of the patients irradiated according to the study protocol. CONCLUSION: Accelerated RT after CT as delivered in this study may increase both acute and long-term morbidity rates compared with studies using standard RT after CT. It did not seem to improve local/regional tumor control or survival despite stringent patient selection criteria. PMID- 8708725 TI - Cisplatin-fluorouracil exclusive chemotherapy for T1-T3N0 glottic squamous cell carcinoma complete clinical responders: five-year results. AB - PURPOSE: To evaluate cisplatin-fluorouracil exclusive chemotherapy (EC) for T1 T3N0 glottic squamous cell carcinoma complete clinical responders (CCR) after cisplatin-fluorouracil induction chemotherapy (IC). PATIENTS AND METHODS: A retrospective analysis was performed of 58 patients with T1-T3N0 glottic squamous cell carcinoma CCR after IC consecutively managed at our department between 1985 and 1992. Twenty-one CCR were managed with EC. Thirty-seven CCR were managed with IC and a conventional laryngeal-preservation modality. Analyses of survival, local control, nodal recurrence, distant metastasis, and metachronous second primary tumor were performed using the Kaplan-Meier actuarial life-table method. In CCR managed with EC, the independent factors of age, tumor classification, exact tumor location, true vocal cord motion, arytenoid cartilage motion, total dosage of drugs delivered, and number of courses received were tested for potential correlation with survival, local recurrence, nodal recurence, and distant metastasis. RESULTS: The 5-year survival, local control, nodal recurrence, distant metastasis, and metachronous second primary tumor rates in CCR managed with EC were 95.2%, 70.7%, 0%, 0%, and 14.3%, respectively. The 5 year rates of survival, local control, nodal recurrence, distant metastasis, and metachronous second primary tumor in CCR managed with IC and a conventional laryngeal-preservation modality were 86.1%, 97%, 2.7%, 6%, and 14.5%, respectively. Local recurrence was statistically more likely in CCR managed with EC (P = .002). Local recurrence in CCR managed with EC was always salvaged with partial laryngectomy or radiation therapy, which resulted in an overall 100% local control and laryngeal-preservation rate within this group. In CCR managed with EC, none of the variables analyzed was statistically related to survival, local recurrence, nodal recurrence, or distant metastasis. CONCLUSION: The present retrospective studies demonstrated that within T1-T3N0 glottic squamous cell carcinoma CCR, there is clearly a significant subset of patients with chemocurable tumors who achieved both perfect preservation of structure supporting voice and long-term survival after EC. Careful monthly follow-up evaluation allowed for timely successful salvage of local recurrence after EC without the need for total laryngectomy. Such management did not appear to increase the risk for subsequent nodal failure, subsequent distant metastasis, or reduced survival. PMID- 8708726 TI - Randomized trial comparing induction chemotherapy versus induction chemotherapy followed by maintenance chemotherapy in small-cell lung cancer. European Lung Cancer Working Party. AB - PURPOSE AND METHODS: The European Lung Cancer Working Party (ELCWP) performed a randomized trial with the primary end point to determine if maintenance chemotherapy with 12 courses of etoposide (120 mg/m2 on days 1 and 3) and vindesine (3 mg/m2 on day 3) could improve progression-free survival in small cell lung cancer (SCLC) patients who responded to six courses of induction chemotherapy with ifosfamide, etoposide, and an anthracycline (doxorubicin or epirubicin). RESULTS: Among 235 eligible patients initially registered, 91 were randomized to receive maintenance therapy, including seven patients who were no longer responding. Among 84 randomized responders, progression-free survival was significantly improved (P = .003) by maintenance therapy, with median durations (maintenance v follow-up) of 25 versus 12 weeks after the second randomization, but survival was not significantly increased (P = .10), with median durations of 48 and 38 weeks. However, in a multi-variate analysis that took into account disease extent, maintenance therapy, Karnofsky performance status (PS), and absolute dose-intensity (ADI) of anthracycline given during induction, limited disease (LD) and maintenance were found to be independent positive predictors of survival. CONCLUSION: We conclude that maintenance chemotherapy in responding patients is beneficial in SCLC. PMID- 8708727 TI - Phase II study of topotecan in patients with extensive-stage small-cell carcinoma of the lung: an Eastern Cooperative Oncology Group Trial. AB - PURPOSE: To determine the response rate and survival of chemotherapy-naive patients with extensive-stage small-cell lung cancer (SCLC) treated with topotecan, and to determine the relationship of topotecan pharmacokinetics with response and toxicity. PATIENTS AND METHODS: Forty-eight patients with previously untreated, extensive-stage SCLC received 2.0 mg/m2 of topotecan daily for 5 days. The first 13 patients were treated without colony-stimulating factor (CSF) support; the next 35 patients received 5 micrograms/kg of granulocyte-colony stimulating factor (G-CSF) for 10 to 14 days starting on day 6. Cycles were repeated every 3 weeks for a maximum of four cycles. Patients who had a partial response to topotecan after four cycles, stable disease after two cycles, or progressive disease at any time received salvage chemotherapy with cisplatin and etoposide. Topotecan pharmacokinetics were measured using a four-point sampling scheme. RESULTS: Of 48 patients, none had a complete response and 19 had a partial response, for an objective response rate of 39% (95% confidence interval [CI], 25.2% to 53.0%). The median response duration was 4.8 months (95% CI, 3.0 to 7.3). After a median follow-up duration of 18.2 months, the overall median survival time was 10.0 months (95% CI, 8.2 to 12.7); the 1-year survival rate was 39% (95% CI, 25.2% to 53.0%). Eight of 34 patients (24%) who received salvage chemotherapy responded. Four of 17 patients who did not respond to first-line therapy with topotecan responded to cisplatin and etoposide. The most common toxicity was hematologic. Ninety-two percent of patients treated without G-CSF developed grade 3 or 4 neutropenia, compared with 29% who received G-CSF. However, the incidence of neutropenic fevers was similar between the two groups (8% and 11%, respectively), and one patient in each group died of neutropenic fevers. There were no differences in objective tumor response, duration of response, time to treatment failure, or survival between the 13 patients who entered the study before G-CSF administration was mandated and the 35 patients who entered after and received G-CSF. There was poor correlation between the WBC count and absolute neutrophil counts (ANCs) and both the area under the curve (AUC) and maximum concentration++ (Cmx) of total topotecan in plasma. There was no correlation between the tumor response and either AUC or Cmx of total topotecan. CONCLUSION: The activity of topotecan in extensive-stage SCLC noted in this study warrants further investigation of this agent in phase III clinical trials. PMID- 8708729 TI - Radiation in management of patients with dermatofibrosarcoma protuberans. AB - PURPOSE: The preferred treatment of dermatofibrosarcoma protuberans (DFSP) is wide resection, namely, margins > or = 3 cm beyond the evident disease and histologically negative margins. We assess the success achieved by radiation combined with surgery for positive/close margins or by radiation alone for those tumors that are not resectable for technical/medical reasons. The literature on this point is virtually nonexistent. MATERIALS AND METHODS: The outcome of treatment of 18 patients with DFSP by radiation alone (n = 3) and radiation and surgery (n = 15) at the Massachusetts General Hospital was assessed. All of the lesions at the time of the treatment by radiation alone or combined with surgery were less than 10 cm. This was the maximum dimension. The actual tumor volume was much less than indicated by this maximum dimension, as the tumors were usually relatively flat. RESULTS: The 10-year actuarial local control rate was determined to be 88%. Local control was realized in the three patients treated by radiation alone, with follow-up periods of > or = 9 years. Among 15 patients treated by radiation and surgery, there have been three local failures; the 10-year actuarial local control rate was 84%. The three local failures occurred in 12 patients whose surgical margins were positive. One of these three local failures developed in the group of two patients whose lesions were scored as grade II. CONCLUSION: Radiation in well-tolerated dose schedules is an effective option in the management of patients with DFSP. This appears to be true for radiation alone or postoperatively for margin-positive disease (primary or recurrent). PMID- 8708728 TI - Randomized phase III trial of liposomal daunorubicin versus doxorubicin, bleomycin, and vincristine in AIDS-related Kaposi's sarcoma. AB - PURPOSE: To compare the safety and efficacy of liposomal daunorubicin (DaunoXome; NeXstar Pharmaceuticals, Inc, Boulder, CO) with a reference regimen of doxorubicin, bleomycin, and vincristine (ABV) in advanced AIDS-related Kaposi's sarcoma (KS). PATIENTS AND METHODS: In a prospective randomized phase III trial, 232 patients were randomized to receive DaunoXome 40 mg/m2 or a combination regimen of doxorubicin 10 mg/m2, bleomycin 15 U, and vincristine 1 mg, administered intravenously every 2 weeks. Treatment was continued until complete response (CR), disease progression, or unacceptable toxicity. RESULTS: Of 232 patients randomized, 227 were treated: 116 with DaunoXome and 111 with ABV. The overall response rate (CR or partial response [PR]) was 25% (three CRs and 26 PRs) for DaunoXome and 28% (one CR and 30 PRs) for ABV. The difference in response rates was not statistically significant. The median survival time was 369 days for DaunoXome patients and 342 days for ABV patients (P = .19). The median time to treatment failure was 115 days for DaunoXome and 99 days for ABV (P = .13). ABV patients experienced significantly more alopecia and neuropathy (P < .0001). DaunoXome patients experienced more grade 4 neutropenia (P = .021). Cardiac function remained stable, with no instances of congestive heart failure on either treatment arm. CONCLUSION: In this large phase III trial, the efficacy of DaunoXome was comparable to that of ABV. Response rates, time to treatment failure, and overall survival were similar on both treatment arms. DaunoXome is a safe and effective primary therapy for advanced AIDS-related KS. PMID- 8708730 TI - Comparative cytotoxicity of dexamethasone and prednisolone in childhood acute lymphoblastic leukemia. AB - PURPOSE: The relative cytotoxicity of prednisolone and dexamethasone in acute lymphoblastic leukemia (ALL) is controversial. We therefore compared the direct antileukemic activities of these compounds in stroma-supported cultures of leukemic lymphoblasts. MATERIALS AND METHODS: Bone marrow samples from children with B-lineage ALL were cultured on allogeneic bone marrow-derived stromal layers and exposed to various concentrations of glucocorticoids. After 4 days of culture, the number of viable leukemic cells was counted by flow cytometry and compared with that in parallel cultures without drugs. RESULTS: In 28 B-lineage ALL samples tested, the concentration producing 50% cytotoxicity (LC50) of prednisolone ranged from 2.0 to 7,978 nmol/L (median, 43.5 nmol/L), and that of dexamethasone from 0.6 to 327 nmol/L (median, 7.5 nmol/L). Despite the wide variability of responses among samples, there was an excellent correlation between LC50 values obtained with the two drugs (linear r = .99, P < .0001; Spearman rank-order r = .77, P < .0001). The median ratio of dexamethasone to prednisolone LC50 and LC90 values was 1:5.5 (range, 1:1.0 to 1:24.4 for LC50; 1:1.1 to 1:25.5 for LC90). Studies with ALL cell lines demonstrated that both drugs were cytotoxic through induction of apoptosis. Stromal layers did not absorb or inactivate measurable amounts of corticosteroids, which indicates that the assay system did not bias results toward increased drug resistance. CONCLUSION: In a bone marrow-derived microenvironment, dexamethasone is five to six times more cytotoxic (on a molar basis) than prednisolone, in agreement with the antiinflammatory activities of these drugs. This finding may serve to guide the selection of dexamethasone dosage in the treatment of ALL. PMID- 8708731 TI - Bronchioloalveolar carcinoma. AB - PURPOSE: To review bronchioloalveolar carcinoma (BAC). MATERIALS AND METHODS: English language articles were identified through a Melvyl Medline Search (1966 to 1995) and through the bibliographies of selected articles. RESULTS: An increase in BAC appears to be responsible for the observed rise in the incidence of adenocarcinoma of the lung. Patients with BAC tend to be younger at diagnosis, are more likely to be female, and less likely to be cigarette smokers when compared with other patients with non-small-cell lung cancer (NSCLC). The etiology of this disease is unclear, but multiple environmental insults have been implicated. There are three subtypes of BAC and the symptoms and prognosis of the disease depend on the subtype and extent of disease, but are generally similar to other histologic types of NSCLC. The radiographic differential diagnosis is broad and includes both benign and malignant diseases. The treatment approach to patients with BAC is similar to those with other types of NSCLC. CONCLUSION: BAC appears to be increasing in incidence, especially in young, nonsmoking females. Three subtypes of the disease exist and are responsible for the variable clinical presentations. Further epidemiologic investigation is needed to elucidate the etiology and pathogenesis of this unique disease. PMID- 8708732 TI - American Society of Clinical Oncology: a society in transition. PMID- 8708733 TI - Tumor size and prognosis in aggressively treated osteosarcoma. PMID- 8708734 TI - Microvascular count and prognosis in colorectal cancer. PMID- 8708735 TI - Cytoreduction and prognosis in childhood acute lymphoblastic leukemia. PMID- 8708736 TI - Relationship of histologic grade, c-erbB-2 expression, and inflammatory infiltrate to prognosis in carcinoma of the breast. PMID- 8708737 TI - Antiestrogens and the reduction in circulating cholesterol. PMID- 8708738 TI - Extended lymph node dissection in gastric cancer induces substantial stage migration and increases stage-specific survival without improvement of overall survival. PMID- 8708739 TI - Long-term response data for 255 patients with metastatic renal cell carcinoma treated with high-dose recombinant interleukin-2 therapy. PMID- 8708740 TI - Effect of beta-lactamase inhibitors on normal immune capabilities and their interactions with staphylococcal pathogenicity. AB - The effects of the beta-lactamase inhibitors, clavulanic acid, sulbactam and tazobactam on normal immune responses were investigated. These agents did not interfere with either humoral or cell-mediated immune responses as measured by the hemolytic plaque assay and delayed type hypersensitivity reaction assay respectively. In addition, human polymorphonuclear leukocyte phagocytic activity was not altered by these agents. When these agents were tested for their effect on Staphylococcus aureas adherence to buccal epithelial cells we found that all inhibitors suppressed staphylococcal adherence at therapeutic serum concentrations. Among the inhibitors investigated, sulbactam was found to significantly inhibit the hemolysin production of S. aureus. These data suggest that beta-lactamase inhibitors do not exhibit immunomodulating activity, but they interfere with some of the virulence factors of S. aureus. These findings suggest the advantage of preparations containing these inhibitors. PMID- 8708741 TI - RP 59500, a streptogramin derivative, is effective in murine listeriosis. AB - RP 59500 (Synercid) a streptogramin derivative, is a mixture of two water-soluble substances, RP 57669 (quinupristin), and RP 54476 (dalfopristin). It was tested in vitro and in vivo against Listeria strains. All strains were sensitive in vitro. The MICs of 60 strains of Listeria monocytogenes, L. seeligeri, L. ivanovii, and L. innocua were between 0.156 and 0.625 mg/l. Strains of L. grayi were inhibited by 1.25 mg/l of RP 59500. In contrast to its bactericidal effect against other gram-positive bacteria, RP 59500 was bacteriostatic against L. monocytogenes in all concentrations tested (up to 16 x MIC). However, it exerted a pronounced postantibiotic effect. RP 59500 was ineffective against intracellular L. monocytogenes multiplying in L929 mouse fibroblast cells. When it was included in the supernatant of the cells in nontoxic concentrations of below 12.5mg/l alone or in combination with gentamicin (50mg/l) it was not able to inhibit the growth of the listeriae in the cells. However, when tested in peritoneal exudate cells, it was bacteriostatic against L. monocytogenes. In spite of its relatively poor effects on listeriae in vitro, RP 59500 was as active as erythromycin in a mouse model of listeriosis. When mice iv infected with L. monocytogenes were treated ip with 2mg/animal every 12 hours with either erythromycin or RP 59500, both substances prevented growth of the bacteria in the organs, but were unable to eradicate the listeriae. We conclude that RP 59500 is a candidate substance for the treatment of human listeriosis which might be used when therapy with ampicillin is not feasible. PMID- 8708742 TI - Ticarcillin/clavulanic acid: determination of minimal inhibitory concentrations against bacterial strains isolated from patients in intensive care units. Comparison with other agents. AB - A total of 303 bacterial strains isolated from bronchoaspirates of Intensive Care Unit (ICU) patients, collected through June and December 1993, were tested for susceptibility to ticarcillin/clavulanic acid, imipenem, amikacin, ceftazidime, ciprofloxacin and piperacillin. The minimal inhibitory concentration (MIC) for each antibiotic was determined according to the NCCLS, by means of serial dilution on microplates. The isolates, 80.8% of which were beta-lactamase producing strains, belonged to Pseudomonas aeruginosa (79 strains), Pseudomonas fluorescens (8 strains), Xanthomonas maltophila (25 strains), Escherichia coli (16 strains), Klebsiella-Enterobacter-Serratia (KES) (62 strains), Proteus spp. (15 strains), Acinetobacter spp. (22 strains), Moraxella spp. (15 strains), Bacteroides catarrhalis (8 strains), Haemophilus spp. (11 strains), Staphylococcus aureus (32 strains), Enterococcus faecalis (10 strains). The highest rate of susceptibility to ticarcillin/clavulanic acid (100%) was detected among E. faecalis (MIC 2-16 micrograms/ml), B. catarrhalis (MIC 1-4 micrograms/ml) and Haemophilus spp. (MIC 1-4 micrograms/ml). Among the non fermenting microorganisms ticarcillin/-clavulanic acid showed good activity toward P. aeruginosa and P. fluorescens (86% and 75% respectively). It was also very active against X. maltophilia with a susceptibility of 96%. Susceptibility to the other antibiotics tested was within the range of 16% and 28%. PMID- 8708743 TI - Piperacillin/tazobactam in the treatment of hospitalized patients with urinary tract infections: an open non-comparative and multicentered trial. AB - The aim of this multicentered, prospective and open study was to determine the clinical and bacteriological efficacy and safety of piperacillin/tazobactam (4g/500 mg IV tid) in the treatment of 79 adult patients with complicated urinary tract infections (UTI) requiring hospitalization. Forty-seven women and 32 men (mean age 54.2 years, and range 21-91) from 4 Argentinean and 6 Mexican hospitals were enrolled. Sixty-one clinically and bacteriologically evaluable patients were treated for a mean of 9.1 days (range 5-15). A favorable clinical response was seen in 83.6% and 80% at early and late assessment, respectively. Bacteriological eradication was achieved in 85.3% and 80% at early and late estimation, respectively. Escherichia coli was isolated in 33 cases, Klebsiella pneumoniae in 8, Enterococcus spp. in 7, Proteus mirabilis in 6, Pseudomonas aeruginosa in 3, Enterobacter spp. and Morganella morganii in 2. While 21% of all the clinical isolates were resistant to piperacillin, none of them was initially resistant to piperacillin/tazobactam. However, one female patient with a persistent UTI caused by E. coli developed resistance to piperacillin/tazobactam during treatment. A 64 year-old man with frontal meningioma developed purulent meningitis due to Enterobacter cloacae after neurosurgery. He was initially treated with ciprofloxacin, rifampin and amikacin and because of persistence of fever, he was moved to piperacillin/tazobactam. After 5 days of therapy, he developed coma secondary to intracranial hemorrhage and died. By then, the platelet count was normal (220,000/microliters), but the prothrombin time (19.5 seconds) and the partial thromboplastin time (63 seconds) were significantly prolonged. Our data suggest that piperacillin/tazobactam is a reliable therapy for complicated, non complicated, community or hospital-acquired UTI. PMID- 8708744 TI - An open, multicenter clinical trial of piperacillin/tazobactam in the treatment of pediatric patients with intra-abdominal infections. AB - A total of 60 children with secondary peritonitis were enrolled in an open, non comparative multicenter study designed to evaluate the safety, tolerance and efficacy of parenteral piperacillin/tazobactam (80/10 mg/kg every 8 hours) in young children. The most common diagnosis was perforated appendicitis (90%) and the three most common pathogens, obtained from the peritoneal cavity, were Escherichia coli (52 isolates), Pseudomonas aeruginosa (16 isolates) and Bacteroides sp. (19 isolates). Patients were examined daily during therapy, 4-14 days and 4-6 weeks post-therapy. Of the 60 patients, 43 were evaluable. The majority of patients had polymicrobial infections (36 patients). All the aerobic isolates were susceptible to piperacillin/tazobactam while 19 were resistant to piperacillin alone. Four of 43 clinically evaluable patients were considered a clinical failure and 3 of 40 bacteriologically evaluable patients were considered to have an unfavorable microbiological response. There were 2 clinically adverse events considered related to the study drug and several possibly related, mild and transitory, abnormalities in eosinophil counts and liver function tests. Based on the safety and efficacy results from this study, the advantages of using a single agent for the treatment of mixed infections of the peritoneal cavity and its potential activity against resistant organisms, we believe that further comparative clinical trials in children with intra-abdominal infections are warranted. PMID- 8708745 TI - Rifabutin as salvage therapy for cases of chronic multidrug-resistant pulmonary tuberculosis in Taiwan. AB - This study was aimed at assessing the efficacy and tolerability of rifabutin for the re-treatment of cases of chronic, multidrug-resistant pulmonary tuberculosis. The study design was self-controlled, single center. Rifabutin was administered as part of an individual-tailored multidrug regimen. In-patients suffering from pulmonary tuberculosis, infected with Mycobacterium tuberculosis bacilli resistant to isoniazid, rifampicin and other drugs with progressive disease unresponsive to prior courses with standard anti-tuberculosis medications were treated. Overall, 43 patients were enrolled and treated with rifabutin at 300 or 450mg/day according to body weight in conjunction with available anti-tuberculous drugs for a mean time of 353 days (range 42-678). Of these, 36 met all eligibility criteria (i.e. positive baseline culture of sputum with bacilli resistant to rifampicin at least) and were retained for the analysis of efficacy. Seventeen patients (47%) achieved a sustained conversion to a negative culture of sputum in a mean time of 47.7 days with a range of 14-120 days. Treatment prevented deterioration in most patients and resulted in clinical and radiological cure or marked improvement in more than half of cases. No correlation was found between treatment outcome and use of medication concomitant to rifabutin or susceptibility of bacilli to the drugs used. Four deaths occurred due to disease progression, in no case being related to study drugs. Ten patients reported a total of 18 adverse events that led to treatment discontinuation in 5 cases. Rifabutin should be considered for inclusion in regimens for cases of pulmonary multidrug-resistant tuberculosis which fail to respond to previous therapy. PMID- 8708746 TI - Comparative study of the cytostatic effects of Oliphen and Ukrain. AB - We evaluated the cytostatic effect of the antihypoxic drug Oliphen (Polyoxiarilene) in comparison with the well known cytostatic and immunomodulating agent Ukrain (a semisynthetic compound derived from Chelidonium majus L. alkaloids). We used tumor cell lines M-HeLa and Hep-2/0-6-5, normal cell cultures from human embryo kidney and lung fibroblasts. The cytodestructive effect of Ukrain on both tumor cell lines was more significant at all tested concentrations. Oliphen was cytotoxic only against Hep-2 but not the HeLa cell line. The proliferative activity of both normal cell cultures decreased in the presence of Oliphen and Ukrain concentrations of 0.1 and 0.015% and was stable at 0.0015 and 0.00015%. PMID- 8708747 TI - Effect of cis-diamminedichloroplatinum (II) on rat thymocyte membrane potential. AB - The effect of cis-diamminedichloroplatinum (II) (cisDDP) on rat thymocytes has been investigated. Changes of anisotropy constant and depolarization of the cell membrane have been determined. The effect of cisDDP on the anisotropy constant has been observed on treated animal thymocytes but not on treated isolated cell membranes. CisDDP inhibits KCl-induced depolarization in isolated rat thymocytes. This inhibiting effect is still present at a concentration as low as 5 microM and it is dose-dependent with a 9.6 microM ED50. CisDDP also boosts the inhibition of the polarization caused by 10 microM tetrodotoxin (TTX) and 10 microM doxorubicin (DOXO). The replacement of Na+ by choline and the inactivation of Ca2+ by EGTA, in the incubation medium, reduces the inhibiting effect of cisDDP on the KCl induced depolarization. These results suggest that cisDDP induced alterations could be due to the interference of cisDDP with the activity of the ionic channels pertaining to Na+ and Ca2+. This interference is stronger for Na+ channels. PMID- 8708748 TI - Treatment of advanced urothelial carcinoma with M-VECA (methotrexate, vinblastine, epirubicin and carboplatin). AB - Transitional cell carcinoma of the urinary tract is actually considered very sensitive to cisplatin-containing regimens. Nevertheless, the generally impaired renal function and poor performance status of these patients are responsible for the severe toxicity usually occurring when cisplatin, either alone or in combination with other agents, is administered to these patients. The aim of this study was to verify the possibility of substituting carboplatin for cisplatin, and epirubicin for doxorubicin in the M-VAC regimen in order to reduce toxicity and improve patient tolerance. Thirty-five patients with advanced urothelial tract carcinoma were treated with a chemotherapeutic regimen composed of methotrexate (30 mg/m2 iv on days 1, 15, 22), vinblastine (3 mg/m2 iv on days 2, 15, 22), epirubicin (35 mg/m2 iv on day 2) and carboplatin (250 mg/m2 iv on day 2) every 4 weeks (M-VECA). All patients had bidimensionally measurable disease. Of the 32 evaluable patients, 5 (16%) obtained a complete response and 10 (31%) a partial response (response rate: 47% C.I. = 30%-64%). Grade III-IV leuko thrombocytopenia was observed in 25%, and mucositis in 19% of cases. Nevertheless, recovery was prompt and opportune dosage reductions avoided severe toxicity in subsequent cycles in most patients. In conclusion, M-VECA is a safe and effective regimen for the treatment of patients with metastatic urothelial tumors. PMID- 8708749 TI - Combined chemotherapy in pleurectomized malignant pleural mesothelioma patients. AB - A phase II clinical trial of 20 cancer patients who presented with malignant pleural mesothelioma (MPM) between November 1991 and April 1993 was conducted. Of the histologically proven cases, 16 (80%) were epitheloid and 4 (20%) were mixed type MPM. Patients were treated with mitomycin C, cisplatin, and alpha interferon after pleurectomy. Our schedule consisted of 10 mg/m2 mitomycin C i.v. infusion, 50 mg/m2 cisplatin i.v. infusion, 10 mil Ur-alpha interferon i.m. and 10 mil Ur alpha interferon i.v. infusion on the first day of treatment. Patients were given a mean of 4.5 chemotherapy cycles (range: 2-6). None of the patients showed complete or partial response. Stable disease was observed in 15 patients, while 5 patients had progressive courses. The overall median survival time after chemotherapy was 12 months (range: 3-31 months). Median survival after chemotherapy was 15 months (range: 4-31 months) in the stable disease group (n:15, 75%), and 5 months (range: 3-13 months) in progressive cases (n:5, 25%). The overall survival rates were 55% [95% Confidence Interval (CI):43%-88.8%] at one year and 15% (95% CI:5%-39.1%) at 2 years. Five patients had grade 3 alopecia, three had grade 2 vomiting and nausea, two had grade 2 leukopenia, one had grade 2 cardiotoxicity and another had discoloration on his fingernails. In our multimodal therapy protocol, we found no difference in survival and relapse rates between our combined modal therapy and other single modal therapies in the literature. PMID- 8708750 TI - Report of Arcanobacterium haemolyticum meningitis in a cancer patient. PMID- 8708752 TI - Anaerobic bacteria in postsurgical infections: isolation rate and antimicrobial susceptibility. AB - Three hundred clinical samples, obtained from post-surgical infections, were evaluated. 37% of samples were positive for anaerobes, 99 samples (33%) for aerobes and anaerobes in mixed culture, and 13 (4%) only for anaerobes. One hundred forty-nine anaerobic strains were isolated: Bacteroides and Clostridia occurred most frequently (34% and 23% respectively). Antimicrobial susceptibility of the isolates was tested by means of a commercial broth microdilution method. In addition, the standardized agar dilution method was performed to evaluate the susceptibility to 8 antibiotics of the 51 Bacteroides strains. PMID- 8708751 TI - First report of Prototheca wickerhami algaemia in an adult leukemic patient. PMID- 8708753 TI - In vitro activity of metronidazole alone and in combination with clotrimazole against clinical isolates of Trichomonas vaginalis. AB - Trichomonas vaginalis vaginitis is generally treated with oral metronidazole. Widespread use of this drug has led, however, to selection of resistant strains. Topical therapy seems appropriate whenever systemic high dosage schedule cannot be adopted in order to overcome resistance. This study was designed to assess the activity of metronidazole alone and in combination with clotrimazole, an imidazole derivative for topical use, against T. vaginalis. Tests were performed employing the antitrichomonas activity of a fixed ratio of metronidazole with clotrimazole (5:1) which has been recently suggested for topical therapy and the checkerboard technique. All tests were carried out under aerobic conditions to maximize T. vaginalis resistance traits. Minimum inhibitory concentrations (MICs) of metronidazole for the 12 strains studied were in the range reported in the literature (0.5-32 micrograms/ml). The interaction of metronidazole with clotrimazole as assessed by the checkerboard technique gave an indifferent outcome with all the strains assayed (FIC = 1-2). The fixed concentration of drugs, however, produced synergism (FIC = 0.5) in 5 of 12 isolates. Spontaneous resistant strains were not selected from T. vaginalis exposed to sub-lethal levels of the drugs or by culturing a large inoculum in the presence of 1, 2, 4 and 8 times the MICs of metronidazole alone or in combination with clotrimazole. These results confirm and extend previous reports highlighting the good in vitro efficacy of the association of metronidazole plus clotrimazole against T. vaginalis. PMID- 8708754 TI - 1996 SNM annual meeting: medical problem solving. PMID- 8708755 TI - In vivo quantification of human pulmonary beta-adrenoceptor density using PET: comparison with in vitro radioligand binding. AB - A new method has recently been developed to quantify pulmonary beta-adrenergic receptors in vivo using PET. This study used in vitro radioligand binding assay (RLBA) as the gold standard to validate in vivo PET measurements. METHODS: Five male patients with lung cancer aged 57 yr (range 42-67 yr) were studied. PET scanning was performed the day before thoracotomy to determine regional pulmonary beta-receptor density. RLBA was carried out on cell membranes prepared from specimens of lung tissue obtained at the thoracotomy to measure beta-receptor density in vitro. In both cases, the hydrophilic nonselective beta-antagonist radioligand (S)-CGP-12177 was used. For PET studies, this was labeled with 11C and for RLBA with 3H. RESULTS: In the PET study, beta-receptor density (Bmax) was 9.43 +/- 1.32 pmole g-1 tissue. In the RLBA study, Bmax was 99.0 +/- 15.5 fmole mg-1 protein, equivalent to 9.90 +/- 1.55 pmole mg-1 tissue. These values are in good agreement with previously reported in vitro measurements on human lung membranes using 125I-iodocyanopindolol. A correlation was found between beta adrenergic density obtained using PET and beta-adrenergic density obtained using RLBA (r = 0.92; p < 0.05). CONCLUSION: The results support the use of PET as a new method for imaging and the way for studies of physiological and pharmacological regulation of beta-adrenergic receptors through noninvasive serial measurements. PMID- 8708756 TI - Overall accuracy of technetium-99m-MAG3 clearance measurements obtained with a gamma camera heart curve. AB - The aim of the present study was to evaluate the accuracy of 99mTc-MAG3 clearance measurements using a precordial gamma camera curve calibrated by a single plasma sample. METHODS: Technetium-99m-MAG3 was administered to ten young normal volunteers. A 60-min gamma camera acquisition was performed. Five different segments of the gamma camera curve were determined: 3 min to 20 min, 3 min to 30 min, 3 min to 40 min, 3 min to 50 min and 3 min to 60 min. A biexponential function was fitted on each of these five different segments, which were thereafter calibrated using eight different blood samples. These blood samples were successively used for calibration at 5, 10, 15, 20, 30, 40, 50 and 60 min. The single injection, multiple plasma sample method was used as reference. RESULTS: Camera clearances varied markedly based on the length of the precordial curve and on the time of the calibration sample. Different regression equations were obtained for each duration of the camera curve, and for each blood sample timing. Correlation coefficients were > 0.95 in most cases recording period of at least 50 min, however, was necessary to obtain a s.e.e. better than those obtained using a single blood sample method without gamma camera curve. CONCLUSION: The 99mTc-MAG3 clearance determination using a gamma camera heart curve calibrated with a single blood sample does not necessarily improve the accuracy of the one blood sample method. PMID- 8708757 TI - Enterogastric bile reflux during technetium-99m-sestamibi cardiac imaging. AB - Enterogastric bile reflux (EGBR), a risk factor for both gastritis and esophagitis, is a potentially treatable noncoronary cause for chest pain. METHODS: To investigate the frequency of EGBR during different 99mTc-sestamibi cardiac imaging, 1405 consecutive 99mTc-sestamibi SPECT myocardial perfusion studies were reviewed. RESULTS: One hundred sixteen of the 1405 patient studies (8.3%) showed EGBR with roughly equal numbers of patients having marked (43 patients), moderate (38 patients) or minimal (35 patients) intensity of abnormal gastric activity. Two examinations showed gastroesophageal reflux of activity. EGBR was less frequent with treadmill stress testing (5.5% patients) than with pharmacologic stress testing using either dipyridamole (11% of patients) or dobutamine (9.2% of patients) (p > 0.005). EGBR also was more frequent in patients over 40 yr of age. Finally, the prevalence of upper gastrointestinal symptoms and the frequency of established upper gastrointestinal diagnoses correlated strongly with the presence and intensity of EGBR. CONCLUSION: Clarification of the full clinical significance of EGBR during 99mTc-sestamibi cardiac imaging is a topic for future research. Nonetheless, the imaging finding of EGBR may, in fact, identify a potentially treatable noncoronary cause for chest pain. PMID- 8708758 TI - Reverse redistribution of technetium-99m-sestamibi following direct PTCA in acute myocardial infarction. AB - A pattern of reverse redistribution (RR) has not been documented in myocardial 99mTc-sestamibi imaging. The purpose of the study was to clarify the time-related changes in myocardial distribution of 99mTc-sestamibi in patients with acute myocardial infarction. METHODS: Myocardial SPECT with 99mTc-sestamibi was performed in 27 patients with acute myocardial infarction within 1 wk after the onset. Twenty-three patients received direct percutaneous transluminal coronary angioplasty (PTCA) and 4 patients did not. Myocardial images were obtained 1 hr (early) and 3 hr (delayed) after the injection of 99mTc-sestamibi. Regional myocardial uptake of 99mTc-sestamibi was scored from 4 (normal) to 0 (no activity), and the RR pattern was defined as a decrease of more than 1 in the regional score at the 3-hr delayed images. Regional myocardial uptake and clearance of 99mTc-sestamibi was also assessed quantitatively. Coronary arteriography and left ventriculography were performed 1 mo later. RESULTS: Out of 22 patients with successful PTCA, RR of 99mTc-sestamibi was observed in 15 patients (68%). Persistent defects (PD) were seen in 12 patients (7 patients with successful PTCA, 1 patient with unsuccessful PTCA, and 4 patients who did not receive angioplasty). In patients with RR, regional uptake of 99mTc-sestamibi in the area of myocardial infarction decreased from 54% +/- 10% in the early images to 43% +/- 8% in the delayed images (p < 0.01). Technetium-99m-sestamibi clearance from the myocardium was faster in the infarct area than in the normal area (26% +/- 7% versus 9% +/- 6%, p < 0.01). Coronary arteriography performed 1 mo later revealed that the patency of the infarct related artery was 100% (15/15) in patients with RR and 50% (6/12) in those with PD (p < 0.01). The extent and severity of a wall motion abnormality were less in patients with RR than in those with PD (extent: 24 +/- 10 versus 36 +/- 9 chord, p < 0.01; severity: -2.7 +/- 0.4 versus -3.4 +/- 0.6 s.d./chord, p < 0.01). CONCLUSION: The RR of 99mTc sestamibi was observed in 68% of patients after successful direct PTCA and was associated with the accelerated clearance of 99mTc-sestamibi from the myocardium. The presence of RR in 99mTc-sestamibi imaging indicates the patency of the infarct-related artery and predicts the preserved left ventricular function. PMID- 8708759 TI - Regional myocardial perfusion assessed with generator-produced copper-62-PTSM and PET. AB - We have previously demonstrated that myocardial perfusion can be estimated accurately in experimental animals with the generator-produced positron-emitting tracer, 62Cu-pyruvaldehyde bis (N4-methylthio- semicarbazone)(62Cu-PTSM) and PET. This study evaluated the feasibility of quantifying regional myocardial blood flow using 62Cu-PTSM and PET in human subjects. METHODS: Regional perfusion was estimated using a previously described and validated two-compartment model from dynamic PET scans obtained after an intravenous bolus of 62Cu-PTSM in 10 healthy volunteers and in 6 patients with coronary artery disease at rest: and in 9 of the volunteers and 4 of the patients after administration of dipyridamole intravenously. Flow estimates were compared with those obtained using H2(15)O. RESULTS: Contrast was high between myocardium and blood or lung with 62Cu-PTSM, resulting in high-quality myocardial images. Liver uptake was also high. At flows of up to 1.5 ml/g/min, flow estimated with 62Cu-PTSM correlated closely with estimates obtained with H2(15)O (y = 0.71x .21, n = 169 regional comparisons, r = 0.66, p < 0.55), but this relationship was not maintained at higher flows. CONCLUSION: The results demonstrate that quantification of myocardial perfusion with 62Cu-PTSM is feasible in human subjects but cannot be used to estimate hyperemic flows due most likely to the strong binding of the tracer to human serum albumin. Copper-62-PTSM congeners with less avidity for human albumin may prove more suitable for evaluation of hyperemic flows. PMID- 8708760 TI - Technetium-99m-sestamibi SPECT to detect restenosis after successful percutaneous coronary angioplasty. AB - This study evaluated the accuracy of 99mTc SPECT in predicting restenosis after primary successful PTCA. METHODS: Thirty-seven patients with equivocal symptom limited exercise stress testing were evaluated. All patients underwent separate day exercise-rest 99mTc-sestamibi SPECT. The perfusion studies were evaluated using three different methods of analysis: visual inspection, semiquantitative and quantitative polar map analysis. The perfusion studies were interpreted in absence of a pre-PTCA scan. All patients underwent a control coronary angiography within 1 mo. RESULTS: Sensitivity and specificity of 99mTc-sestamibi SPECT in predicting restenosis were 87.5-78%, 50-65% and 75-74% for visual inspection, semiquantitative and quantitative polar map analysis, respectively. Sensitivity and specificity related to the vascular territories were: LAD territory 93-73% (qualitative analysis), 53-60% (semiquantitative analysis), 80-67% (quantitative analysis); LCX territory 83-100% (qualitative analysis); and 33-100% (semiquantitative analysis), 67-100% (quantitative analysis); and RCA territory 67-80% (qualitative analysis), 67-60% (semiquantitative analysis), 67-80% (quantitative analysis). CONCLUSION: These data suggest that 99mTc-sestamibi SPECT is a useful noninvasive tool in the follow-up evaluation of patients who have undergone angiographically successful coronary angioplasty even in the absence of a pre-PTCA perfusion study. PMID- 8708761 TI - Simultaneous perfusion tomography and radionuclide angiography during dobutamine stress. AB - The purpose of this investigation was to evaluate the changes in left ventricular function and volumes concurrently with tomographic myocardial perfusion during dobutamine infusion. METHODS: Ninety-two patients underwent first-pass radionuclide angiography using a multicrystal gamma camera and myocardial tomography after high-dose (40 micrograms/kg/min) dobutamine infusion and 99mTc sestamibi administration. RESULTS: Dobutamine increased systolic blood pressure (p < 0.0001), heart rate (p < 0.00017), left ventricular ejection fraction (p = 0.0001), cardiac output (p = 0.0001) and stroke volume (p = 0.042). The end diastolic (p = 0.009) and end-systolic volumes (p = 0.0007) significantly decreased. Of 38 patients with cardiac catheterization, 28 had significant coronary artery disease and 10 had normal coronaries. The sensitivity and specificity for coronary artery disease detection by myocardial perfusion tomography were 78% and 90%, respectively. By radionuclide angiography, only 9 of 27 coronary artery disease patients experienced deterioration of wall motion during dobutamine (sensitivity 33%). CONCLUSION: Changes in myocardial perfusion are more sensitive than changes in left ventricular function for detecting coronary artery disease during dobutamine stress. PMID- 8708762 TI - Pleural effusion and ventilation/perfusion scan interpretation for acute pulmonary embolus. AB - This study was conducted to determine if pleural effusion size affects ventilation/perfusion (V/Q) scan interpretation algorithms for acute pulmonary embolus (PE). METHODS: Retrospective analysis identified 163 consecutive patients undergoing angiography for PE with radiographic evidence for pleural effusion. V/Q scanning was performed in 94 (58%) of cases and reported using original Prospective Investigation of Pulmonary Embolism Diagnosis (PIOPED) criteria. Effusions were classified as small, large and/or bilateral. Radiographic and scintigraphic results were compared with regard to size and location of abnormalities. RESULTS: Of the 163 patients, 57 (35%) had angiographically-proven PE, 77 (47%) had at least one large pleural effusion and 86 (53%) had a small effusion; 33 (43%) with large effusions and 24 (28%) with small effusions had emboli at angiography. Thirty-six of 119 patients (30%) with clear chest radiographs (a control group) had PE. Thus, large effusions were associated with a higher incidence of PE than those with small effusions or clear lungs (p < 0.05). Of those with V/Q scanning, 26 of 94 (28%) had a solitary large effusion, with 12 (46%) positive for emboli. V/Q-matched abnormalities limited to effusion size were found in 16 with a solitary large effusion and 10 with a solitary small effusion. In both groups, 50% were angiographically positive for emboli. Twenty three (66%) of 35 with bilateral effusions had corresponding V/Q-matched defects at one (n = 11) or both (n = 12) lung bases, and 9 (39%) were positive for emboli. In total, 45% with a V/Q-matched defect of equivalent size to the effusion were angiographically positive for PE. CONCLUSION: Pulmonary emboli are associated with pleural effusions of all sizes. Matched V/Q defects corresponding to radiographically-evident pleural effusions are of intermediate probability for PE. Thus, revision of the traditional lung scan interpretive criteria based upon pleural effusion size is not warranted. PMID- 8708763 TI - Small perfusion defects in suspected pulmonary embolism. AB - The purpose of this investigation was to assess the diagnostic value of 1 to 3 versus > 3 small subsegmental defects on perfusion lung scans of patients with suspected acute pulmonary embolism (PE). METHODS: Data from the Prospective Investigation of Pulmonary Embolism Diagnosis (PIOPED) were evaluated from patients with suspected acute PE. Angiograms, follow-up data and outcome classifications were used to determine PE status. The perfusion scan of included patients showed only small subsegmental defects ( < 25% of a segment) in the presence of a regionally normal chest radiograph. Findings on the ventilation scan were irrelevant. RESULTS: The positive predictive value for PE of perfusion lung scans with 1-3 small subsegmental defects was 1% to 3%, depending on the group analyzed. The positive predictive value for the PE of perfusion lung scans with > 3 small subsegmental defects was 11% to 17% depending on the group analyzed. CONCLUSION: Perfusion lung scans with 1-3 small subsegmental defects satisfy the criterion for a very low probability ( < 10% positive predictive value) for PE and perfusion lung scans with > 3 small subsegmental defects satisfy the criteria for a low probability ( < 20% positive predictive value) for PE. PMID- 8708765 TI - Hepatobiliary kinetics of technetium-99m-IDA analogs: quantification by linear systems theory. AB - A compartmental model describing the extraction and disposition of 99mTc acetanilidoiminodiacetic acid (IDA) compounds by the liver has been applied to 5 adult patients admitted for cholecystitis investigations and 29 jaundiced infants the majority of whom were clinically differentiable into neonatal hepatitic and biliary atretic groups. METHODS: In each case kinetic rate constants were calculated to describe hepatocyte extraction of 99mTc-IDA structural analogs from blood pool (k21) and subsequent elimination (k3) of this compound into biliary tract. Also modeled was the reverse-binding constant (k12) describing the return of such radiotracer to the systemic circulation and the blood fraction (f) which accounted for the composite vasculature forming a matrix in the liver. RESULTS: It was shown that these indices could be used to determine accurate compartmental mean residence times (MRT(c)s) for each patient by correlation with values obtained by deconvolutional analysis and independent measurement of leading edge parenchymal transit times. For the adult patients the following indices, typical of good hepatocyte function, were derived: k21 = 0.933 +/- 0.488 min-1, k12 = 0.0277 +/- 0.0340 min-1, k3 = 0.1610 +/- 0.0531 min-1, f = 0.3519 +/- 0.3048 and MRTc = 11.19 +/- 3.13 min. Analysis of the pediatric group revealed no significant differences in their respective MRT(c)s. However, significant differences in the extraction (p < 0.01) and excretion (p < 0.001) coefficients were prominent. CONCLUSION: This method can be applied to provide accurate and meaningful intercompartmental rate parameters and MRT(c)s for adults, nonobstructed and obstructed infants. PMID- 8708764 TI - Diagnostic value of the gallium-67 pulmonary leak index in pulmonary edema. AB - We studied the value of a noninvasive, bedside, dual-radionuclide method (67Ga circulating transferrin and 99mTc-red blood cells) to measure pulmonary microvascular permeability in efforts to discriminate between pulmonary edema due to adult respiratory distress syndrome (ARDS) and hydrostatic pulmonary edema (HPE). METHODS: Patients had respiratory insufficiency and bilateral alveolar pulmonary edema on chest radiographs. All patients, except one, were mechanically ventilated. Patients, were divided into groups according to various sets of etiologic, hemodynamic and ventilatory factors. Group 1 (n = 8) had risk factors for HPE only. Group 2 (n = 5) had risk factors for both ARDS and HPE, such as a pulmonary capillary wedge pressure (PCWP) above 18 torr. Group 3 (n = 13) had risk factors for ARDS only and a PCWP below 18 torr. Patients were also classified on the basis of a lung injury score, using radiographic and ventilatory variables. Group 4 (n = 12) had a score below 2.5 and Group 5 (n = 14) above 2.5, arbitrarily defined as ARDS. Any radioactivity measurements over the lungs and in blood within 72 hr after admission were used to calculate the 1 hr pulmonary leak index as a measure of microvascular permeability (upper limit of normal 14.1 x 10(-3).min-1). RESULTS: The PLI ( x 10(-3).min-1) was median 10.2 (range 4.4-16.2) in Group 1, 26.8 (14.2-31.9) in Group 2 and 32.3 (23.0 52.4) in Group 3 (p < 0.001). It was 13.3 (4.4-39.9) in Group 4 and 31.1 (14.2 52.4) in Group 5 (p < 0.01). Using the various definitions, the sensitivity of a supranormal pulmonary leak index for ARDS was 100% and the specificity varied between 46% and 75%. In receiver operating characteristic curves, the pulmonary leak index performed best when ARDS and HPE were defined on the basis of risk factors only, and performed better than hemodynamic and equally well as ventilatory variables in discriminating between edema types, if definitions of the latter were mainly based on hemodynamic and ventilatory variables, respectively. CONCLUSION: The 67Ga pulmonary leak index is a useful tool to differentiate ARDS from HPE. PMID- 8708766 TI - Biokinetic behavior of technetium-99m-DMSA in children. AB - After intravenous administration of 99mTc-DMSA, biokinetic data were collected from studies on 24 children aged from 5 wk to 14.8 yr (15 normal and 9 with renal pathology). METHODS: Patients were imaged with a gamma camera up to 30 hr postinjection and the absolute activities in the kidneys, liver, spleen, bladder, knees and whole body were estimated using an attenuation-corrected conjugate counting technique. Renal uptake and elimination rates and urinary excretion of radioactivity were also measured. RESULTS: In children with normal renal function, maximal kidney uptake was 42.4% +/- 5.4% and was taken up with a half time of 1.0 +/- 0.2 hr. Renal excretion amounted to 18.0% +/- 4.4% at 24 hr and was lowest in children aged less than 1 yr. In children with abnormal renal function, apart from the expected reduction in renal uptake there was evidence of wider variations in uptake rate and increased urinary excretion. Mean uptakes in liver and spleen were approximately 5% and 2%, respectively, in all patients and uptake in knees, assumed to reside in the metaphyseal growth complexes, was 1.4%. CONCLUSION: In children with normal renal function, there was little evidence of age-dependent biokinetic factors other than reduced urinary excretion and lower uptake in knees in children aged less than 1 yr. The results therefore suggest that a single biokinetic model may suffice for radiation dosimetry purposes in normal children irrespective of age. PMID- 8708767 TI - Radiation dosimetry of technetium-99m-DMSA in children. AB - Radiation dosimetry was performed on 24 children (aged 5 wk to 14.8 yr) who were undergoing routine diagnostic investigation of renal impairment with 99mTc-DMSA. METHODS: Organ doses were calculated using MIRDOSE 3 with biokinetic data obtained in previously described studies, and effective doses and effective dose equivalents were estimated. Interpolation by inverse weight between pediatric anthropomorphic phantoms was compared with age-matching to discrete phantoms. Administered activities were scaled by body surface area from the adult activity of 100 MBq and the resulting radiation doses in normal children were compared with those that would have resulted from a schedule based on body weight. RESULTS: The effective doses estimated by interpolation differed by up to 46% from those based on discrete phantoms and showed less variation. In children with normal bilateral renal function, the mean effective dose per administered activity was 0.91 +/- 0.08 mSv or 0.98 +/- 0.29 mSv by the two methods, respectively. Renal pathology reduced the effective dose, on average, by 15% of the value for normal patients. CONCLUSION: Over the pediatric age range, the uniformity of effective dose values was improved by scaling the administered activity according to body surface area rather than to body weight. PMID- 8708768 TI - Iodine-123-metaiodobenzylguanidine myocardial imaging in patients with right ventricular pressure overload. AB - Iodine-123-metaiodobenzylguanidine ([123I]MIBG) has been used to evaluate the cardiac sympathetic nervous system, particularly that of the left heart. To clarify whether the right ventricular (RV) sympathetic neuronal function could be evaluated by [123I]MIBG myocardial imaging, we applied the technique in patients with pulmonary hypertension that was associated with either chronic pulmonary diseases or pulmonary vascular diseases. METHODS: All patients underwent right heart catheterization, and right heart hemodynamics were determined during a clinically stable state. SPECT was performed in the resting state 15 min (early imaging) and 4 hr (delayed imaging) postadministration of [123I]MIBG. Seven regions of interest (ROI) were selected on the delayed short-axis images on the RV free wall, left ventricular (LV) free wall and interventricular septum (IVS). We calculated the IVS-to-LV uptake ratio from the scintillation counts of the ROI. Thallium-201 myocardial imaging was also performed within 1 wk after [123I]MIBG imaging. RESULTS: Images obtained with these techniques were analyzed for the RV-to-LV uptake ratio. The IVS-to-LV ratio on [123I]MIBG correlated negatively and significantly with the mean pulmonary arterial pressure (PAm). The RV-to-LV uptake ratio on 201Tl images correlated significantly with PAm. CONCLUSION: Our results suggest that the uptake ratio of [123I]MIBG in the IVS is a useful index for evaluating the severity of pulmonary hypertension, and that chronic RV pressure overload contributes to disturbances of the cardiac sympathetic nervous system. PMID- 8708769 TI - Renal technetium-99m-DMSA SPECT in normal volunteers. AB - Dimercaptosuccinic acid (DMSA) renal scintigraphy today is considered a sensitive and useful technique for the detection of cortical abnormalities. Recent studies have suggested that lesion detection can be improved by SPECT imaging. This study investigated normal kidneys using different SPECT modalities. METHODS: Ten young, healthy volunteers with normal clinical history and normal renal ultrasound underwent planar and SPECT DMSA imaging 2 to 4 hr after intravenous injection of 99mTc-DMSA (185 MBq). Analysis of SPECT data was focused on the homogeneity of cortical uptake (comparison of upper and lower pole activity) as well as on the presence or absence of focal cortical defects. RESULTS: No abnormality could be observed on the planar images. SPECT revealed, in seven kidneys (five left and two right), the presence of a hypoactive upper pole. This was visually observed on the coronal slices with up to 35% difference between upper and lower pole. Moreover, three focal cortical defects were visualized on the coronal slices as well as on three-dimensional surface shade displays. CONCLUSION: These normal patterns should be recognized when evaluating a patient with possible renal involvement. PMID- 8708770 TI - Technetium-99m-DMSA renal SPECT in diagnosing and monitoring pediatric acute pyelonephritis. AB - This study compares the sensitivity of 99mTc-dimercaptosuccinic acid (DMSA) renal SPECT with planar scintigraphy, concluding the importance of 99mTc-DMSA renal SPECT for the early diagnosis of acute pyelonephritis (APN) in patients under 3 yr of age. METHODS: Twenty-seven children under 3 yr of age, with clinical and/or laboratory suspicion of APN, were investigated. All 99mTc-DMSA renal SPECT and planar images and voiding cystoureterogram (VCUG) were obtained within 3 days of hospitalization. RESULTS: In the first examination, renal cortical defects were detected in 23 patients (42 kidneys) with SPECT and in 9 patients (11 kidneys) with planar scintigraphy. One year after treatment, constant renal cortical lesions were observed in 11 patients (14 kidneys) with SPECT and 4 patients (4 kidneys) with planar scintigraphy. The high grades of vesicoureteral reflux (VUR) (grade >or= 3) correlate better with APN diagnosed by SPECT (34 kidneys) than by planar scintigraphy (8 kidneys). Multiple renal cortical defects (number of lesions >or= 4) were only seen in patients under 1.5 yr old and none of those with a negative 99mTc-DMSA renal SPECT had a positive 99mTc-DMSA renal planar scintigraphy at any time. There is a significant difference (p < 0.05) between the diagnostic ability of these two methods of examination. CONCLUSION: Our results suggest that 99mTc-DMSA renal SPECT should be used, where possible, instead of planar DMSA in routine examination of children with clinical suspicion of APN, especially for those under 3 yr of age. PMID- 8708771 TI - SPECT bone scintigraphy of anterior cruciate ligament injury. AB - This retrospective analysis of SPECT bone scans of the knee was undertaken to define typical bone scan appearances and to assess the sensitivity of this method. We looked at 14 patients, mostly with chronic knee pain, who had anterior cruciate ligament (ACL) tears detected by MRI. METHOD: Of the 14 patients, 10 were referred for bone scanning following injury and 4 complained of chronic knee pain without injury. Planar scans were performed 4 hr after the injection of 750 MBq of 99mTc-MDP. Tomographic images were obtained by a 64 x 20-sec acquisition over 360 degrees using a high-resolution collimator. MRI imaging included axial and sagittal, T1 weighted and coronal fast field echo (FFE) sequences. Ten patients also had arthroscopy performed. RESULTS: MRI scans showed 6 lone ACL tears and 8 combined with other ligamentous injuries. SPECT scans showed abnormalities in 10 patients in the region of ACL insertions but only 4 planar studies were abnormal. SPECT identified focal activity at the upper (n = 8) or lower (n = 2) insertion of the ACL. Six of 10 arthroscopies confirmed ACL tears, 2 complete and 4 partial. Overall, agreement was found with MRI in 10 of 14 cases and in 8 of 10 with arthroscopy. Abnormalities were identified in 10 of 11 regions of other ligament or bone injury identified by MRI. CONCLUSION: SPECT bone scanning of the knee is superior to planar imaging in detecting ACL injury and is a sensitive technique. Focal activity may be seen at either end of ACL attachment but more commonly at the upper femoral insertion. Knee SPECT may be a valuable examination in suspected ACL injury, particularly if MRI is not available, is equivocal or where clinical signs are absent. PMID- 8708772 TI - Pediatric gastric emptying: value of right lateral and upright positioning. AB - Gastroesophageal reflux and gastric emptying are usually assessed simultaneously with a 1-hr procedure. After ingestion of radiolabeled formula sequential images are gathered when the infant is in the supine position. This position is adequate for gastroesophageal reflux assessment, but delays gastric emptying. METHODS: We studied 48 children, 1 wk to 2 yr of age, who presented with vomiting or failure to thrive. They received 99mTc-sulphur colloid in formula. After completing 1 hr supine imaging we obtained additional abdominal views after changing the position of the infant to right lateral for 30 min, and upright for another 30 min. RESULTS: The percent of gastric emptying at 60 min in the supine position was 35% +/- 19%. After 90 min, in the right lateral decubitus, the percent gastric emptying was 60% +/- 25%. At 120 min, after an upright period, the gastric emptying was 73% +/- 20%. In the supine position 19 of 48 patients showed significant emptying (defined as > 40% emptying). This increased to 41 of 48 normal studies considering the right lateral position and to 45 of 48 normal studies considering the infant upright position. CONCLUSION: Many patients with delayed gastric emptying show significant emptying just by changing position. We routinely complement gastric emptying studies with delayed views in the right lateral and upright position. PMID- 8708773 TI - Pseudo pulmonary embolism in complex congenital heart disease. AB - Two children (aged 12 and 14 yr) with a history of complex congenital heart disease presented with symptoms and signs suggestive of pulmonary embolism. Initial ventilation-perfusion lung scans showed normal, approximately equal ventilation to both lungs. Global reduction of perfusion to the right lung was observed in one and was observed in the left lung in the other patient. It was not possible to exclude a large, central embolus in either case. Due to complex cardiac anatomy, however, which included bilateral cavopulmonary anastomoses in the first patient and SVC-right pulmonary artery and right atrial-pulmonary artery anastomoses in the second, repeat scans were performed within a short period in each case with different injection sites, including the contralateral arm and a leg injection, respectively. In both patients, these follow-up scans showed a different perfusion agent distribution to each lung when compared to the initial scans. These results demonstrated that there was no evidence of pulmonary embolism in either case. PMID- 8708774 TI - Iodine-123-MIBG imaging in pheochromocytoma with cardiomyopathy and pulmonary edema. AB - We encountered a patient with pheochromocytoma associated with a catecholamine induced cardiomyopathy that developed recurrently bilateral and unilateral pulmonary edema. The diagnosis of pheochromocytoma was made by elevated plasma catecholamine levels and the intense tumor [123I]MIBG uptake and was confirmed at the time of surgery. The patient showed reduced myocardial [123I]MIBG uptake with left ventricular dysfunction, and endomyocardial biopsy findings were consistent with the diagnosis of catecholamine-induced cardiomyopathy. After tumor resection, plasma levels of catecholamine were normalized, and pulmonary edema never recurred, although cardiac dysfunction did not show an improvement on echocardiography. Myocardial and lung [123I]MIBG uptake increased when compared to uptake levels on preoperative scans, but myocardial uptake was still below normal. These findings indicated that over-secreted catecholamines influenced both the heart and lungs. Pheochromocytoma can induce cardiac and lung injuries, and [123I]MIBG scanning may contribute not only to tumor characterization but also to assessing and monitoring the influence of catecholamines on the heart and lungs. PMID- 8708775 TI - Focal liver hyperplasia in Alagille syndrome: assessment with hepatoreceptor and hepatobiliary imaging. AB - A child with Alagille syndrome, characterized by intrahepatic bile duct paucity, developed severe liver cirrhosis and was referred for liver transplantation. In the pre-transplantation evaluation, scintigraphic scans were performed using 99mTc-galactosyl serum albumin (99mTc-GSA) as a hepatoreceptor binding agent and 99mTc-pyridoxyl-5-methyl-tryptophan (99mTc-PMT) as a hepatobiliary agent. These studies demonstrated severe hepatobiliary dysfunction with an area of increased focal uptake in the liver. Histological examination at surgery confirmed that this focal lesion was an area of compensatory hyperplasia in advanced biliary cirrhosis. We present the usefulness of these tracers for detecting the focal hyperplasia of the liver. PMID- 8708776 TI - Radionuclide captopril renography in postpartum renal artery aneurysms. AB - We report the case of a young woman with three left renal artery aneurysms, diagnosed immediately postpartum, associated with a de novo high blood pressure. To assess anatomical and functional characteristics of renal artery aneurysms, renal angiogram, MRI, intravenous pyelography, ultrasonography and radionuclide renography were performed. Two patent saccular renal artery aneurysms were demonstrated in the left kidney by renal angiogram. A larger, thrombosed aneurysm was also depicted on the left side on ultrasonography, MRI and renal angiogram. The larger aneurysm was responsible for renovascular disease of the middle third of the kidney, as demonstrated by captopril and baseline radionuclide renographic studies. It also impeded drainage of the lower pyelocalyceal group, without obstructing it, as shown by concomitant furosemide (Lasix) evaluations. PMID- 8708777 TI - Bone scintigraphy in hungry bone syndrome following parathyroidectomy. AB - A 59-yr-old man with chronic renal failure was admitted for evaluation of generalized skeletal pain and frontal bone mass, which was lytic on radiography. Bone scintigraphy demonstrated several foci of moderately increased uptake, without involvement of the skull mass. Radiographs of these lesions were compatible with brown tumors. Serum parathormone level was elevated and CT demonstrated a lower right cervical mass, consistent with parathyroid tumor. Following the removal of the mass and decrease in parathormone levels, the patient suffered from a prolonged period of hypocalcemia and his bone pain worsened. Repeat bone scintigraphy showed an increase in the number and intensity of the areas of focal uptake, consistent with hungry bone syndrome. This flare-up phenomenon is due to an increase in bone metabolism and is an uncommon finding following parathyroidectomy for primary hyperparathyroidism. PMID- 8708778 TI - Blood-pool imaging using technetium-99m-labeled liposomes. AB - This study evaluated two 99mTc-liposome formulations as potential blood-pool agents in comparison with standard 99mTc-red cells and 99mTc-human serum albumin (HSA). METHODS: Liposomes with no surface modification or coated with polyethylene glycol (PEG) were labeled with 99mTc using the lipophilic chelator, HMPAO. Autologous red cells were labeled with 99mTc using in vitro or in vivo techniques. Technetium-99m-HSA was supplied commercially. Rabbits were injected intravenously with 99mTc-liposomes, 99mTc-red cells or 99mTc-HSA. Static images were acquired and blood samples collected. RESULTS: Technetium-99m-liposome images showed prominent blood-pool activity compared to lung and liver activities, which were similar to those acquired for 99mTc-red cells, but better than 99mTc-HSA. Heart-to-lung ratios were not significantly different between 99mTc-liposome formulations or for either formulation compared to 99mTc-red cells. The ratios were higher, however, than for 99mTc-HSA. Heart-to-liver ratios were higher for PEG 99mTc-liposomes than they were for neutral 99mTc-liposomes and 99mTc-HSA, but were not significantly different than 99mTc-red cells. Bladder activities for both 99mTc-liposome formulations were 3-6 times lower than for the other agents. PEG 99mTc-liposomes remained in circulation 1.6 times longer than any of the other agents. CONCLUSIONS: Technetium-99m-liposomes, independent of surface modification, had excellent circulation persistence and in vivo stability when compared to 99mTc-red cells and 99mTc-HSA. PEG 99mTc-liposomes performed better than neutral 99mTc-liposomes due to lower liver background activity. Advantages of PEG 99mTc-liposomes compared to 99mTc-red cells include: (a) only one venipuncture, (b) little exposure to patient's blood, (c) excellent in vitro and in vivo stability and (d) lack of drug interference. PMID- 8708779 TI - Biochemical modification of streptavidin and avidin: in vitro and in vivo analysis. AB - The high affinity streptavidin (or avidin)/biotin system is being investigated for imaging and radiotherapy procedures. Streptavidin (SA) and avidin exhibit markedly different pharmacokinetics, with avidin clearing from the blood much faster than SA. To optimize blood clearance kinetics, SA and avidin were biochemically modified and analyzed in vitro and in vivo. METHODS: Galactose moieties were covalently attached to promote binding by hepatocyte galactose receptors and hasten SA clearance. To prolong avidin clearance, avidin was deglycosylated and/or neutralized by acetylation of its lysine amino acids. In vitro, the modified proteins were analyzed by isoelectric focusing, SDS polyacrylamide electrophoresis and a biotin binding saturation assay. The modified and native proteins were radiolabeled with 131I and injected into rabbits for pharmacokinetic, redistribution and imaging analysis. RESULTS: For SA, the resulting increase in blood clearance and liver accumulation was correlated to the amount of galactose bound to SA. For avidin, each type of modification increased its circulation time, with the slowest clearance resulting from a combination of deglycosylation and neutralization. CONCLUSION: Biochemical modification of SA and avidin resulted in altered pharmacokinetics compared to the native proteins. Modified SA or avidin, when cross-linked with a lesion specific targeting agent, may be applicable for rapid two-step in vivo imaging techniques. PMID- 8708780 TI - Prevention of radiolysis of monoclonal antibody during labeling. AB - Monoclonal antibody may undergo loss of immunoreactivity due to radiation damage when labeled with large amounts of 131I or 90Y for therapy. Our aim was to develop a method to protect an antibody during the labeling procedure. METHODS: As a model we used T101, a murine monoclonal antibody directed against CD5 antigen. Iodine-125-T101 (100 micrograms, 1 ml) was mixed with 90Y-DTPA (0.64 MBq to 165.9 MBq) for 24 hr in order to deliver doses of 5 Gy to 1280 Gy to the solution. In separate experiments, 125I-T101 solutions were irradiated with 60Co external beam delivering radiation doses of 40 Gy to 1280 Gy. The effect of radiation on T101 immunoreactivity was tested by using the CCRF-CEM cell line, and the bound T101 radioactivity was determined. In a final experiment, we directly labeled a DTPA conjugated T101 using 561 MBq of 90Y under conditions delivering approximately 640 Gy to the solution. Previously used radioprotectants including human serum albumin, cysteamine and glycerol were evaluated. We focused on ascorbic acid because it is an FDA approved drug that does not interfere with the radiolabeling process. RESULTS: The immunoreactivity of 125I-T101 was approximately 83%, but at 640 Gy the immunoreactivity decrease to 7%. In contrast, in the presence of radioprotectants this decrease could be abrogated. External irradiation also showed a dose dependent decrease in immunoreactivity to as low as 0.3% at 1280 Gy. Adding ascorbic acid (5.5 mg/ml) to the solutions prior to the irradiation largely abrogated this decrease. The immunoreactivity of T101 labeled with 90Y without protectant showed 46% immunoreactivity whereas, in presence of ascorbic acid (11 mg/ml) full retention of immunoreactivity was observed. CONCLUSION: Various radioprotectants can successfully prevent the loss of immunoreactivity or breakdown as a result of radiolysis. Ascorbic acid is an effective radioprotectant that can be used to prevent loss of antibody immunoreactivity during the labeling process. PMID- 8708781 TI - Inhibition of renal uptake of indium-111-DTPA-octreotide in vivo. AB - Indium-111-DTPA-octreotide has been successfully used for imaging of somatostatin receptor-positive lesions. However, significant renal uptake of 111In-DTPA octreotide exists, reducing the scintigraphic sensitivity for detection of small tumors in the perirenal region and the possibilities for radiotherapy. The aim of the present study was to determine whether renal uptake of 111In-DTPA-octreotide could be reduced in vivo in rats. METHODS: Male Wistar rats (200-250 g) were placed in metabolic cages and injected with 111In-DTPA-octreotide (0.2 MBq and 0.5 microgram octreotide), in the presence or absence of re-uptake blockers. At time t = 20 hr after injection, rats were sacrificed and organs were isolated and counted for radioactivity. RESULTS: Adding NH4Cl or NaHCO3 to the food, resulting in the production of more acid or alkaline urine respectively, resulted in less radioactivity in the kidneys 20 hr after injection compared to controls. Lysine in a single dose of 400 mg/kg resulted in an inhibition of kidney uptake of 40%. When lysine was injected 30 min before 111In-DTPA-octreotide, the inhibition was 25%. Arginine had less effect on tubular uptake of 111In-DTPA-octreotide than lysine (20% inhibition). Sodium maleate inhibited kidney uptake of 111In-DTPA octreotide most successfully. Acetazolamide (100 mg/kg), succinylacetone (100 mg/kg), cystine dimethylester (340 mg/kg) and increase in urinary flow did not influence 111In-DTPA-octreotide retention in the kidneys. CONCLUSION: It appeared possible to reduce re-uptake of 111In-DTPA-octreotide in the rat kidney in vivo. The most pronounced effects were seen after administration of sodium maleate or lysine but, because of the described toxic effects of maleate, we will study further only the effects of lysine in a clinical setting. PMID- 8708782 TI - Detecting infection and inflammation with technetium-99m-labeled Stealth liposomes. AB - The performance of 99mTc Stealth liposomes was investigated in various rat models. METHODS: Preformed polyethyleneglycol-containing liposomes with encapsulated reduced glutathione, were radiolabeled using the lipophilic 99mTc HMPAO. The labeled liposomes were intravenously administered to rats with focal S. aureus or E. coli infection, or turpentine-induced inflammation. For comparison, Tc-99m-nanocolloid- and 99mTc-labeled nonspecific IgG were tested. In rats with Pneumocystis carinii pneumonia (PCP), Tc-99m-liposomes were directly compared to In-111 labeled nonspecific IgG. RESULTS: Technetium-99m-liposomes accumulated in the infectious and inflammatory muscle foci over 24 hr (0.59% injected dose per gram tissue (%ID/g) for S. aureus; 1.18 %ID/g for turpentine). Abscess-to-muscle ratios increased to values as high as 24.0, 41.7 and 44.5 for the respective models at 24 hr postinjection. Technetium-99m-liposomes visualized the foci as early as 1 hr postinjection. Technetium-99m-IgG visualized S. aureus infection, but abscess-to-muscle ratios and abscess uptake at the later time points were significantly lower. Technetium-99m-nanocolloid failed to visualize any of the muscle foci. In PCP however, 99mTc-liposomes did not show preferential localization in the infection. The control agent 111In-IgG showed a significant, two-fold increase in lung uptake. CONCLUSION: Technetium-99m-Stealth liposomes preferentially accumulated in abscesses, leading to very high target-to-nontarget ratios. This property appears to be related to a process based on uptake of long circulating particles. In a specific type of infection, i.c. PCP, 99mTc-liposomes did not accumulate in diseased lung tissue, thus mimicking the in vivo behavior of labeled leukocytes. PMID- 8708783 TI - Comparison between thallium-201 and technetium-99m-tetrofosmin uptake with sustained low flow and profound systolic dysfunction. AB - Technetium-99m-tetrofosmin uptake was compared to that of 201Tl in the setting of low flow and systolic dysfunction. METHODS: In nine open-chested dogs, a severe left anterior descending (LAD) coronary artery stenosis resulted in a 54.3% mean flow reduction and decreased left ventricular thickening from 21% +/- 1% to -3 +/ 2%. After 30 min, 37 MBq (1 mCi) of 201Tl and microspheres were injected and initial and 2-hr redistribution images acquired. Two hours later, 370 MBq (10 mCi) of 99mTc-tetrofosmin and microspheres were injected and an image was obtained. LAD: left circumflex (LCX) count ratios for both tracers and flows were calculated by well counting postmortem, and 201Tl and 99mTc-tetrofosmin defect magnitudes were determined by quantitative image analysis. RESULTS: LAD:LCx flow ratios were similar during 201Tl and 99mTc-tetrofosmin injections (0.48 +/- 0.04 versus 0.49 +/- 0.05, p = n.s.). Final 201Tl activity (0.66 +/- 0.04) was significantly higher than 99mTc-tetrofosmin (0.55 +/- 0.05; p < 0.05). LAD/LCx 99mTc-tetrofosmin image defect count ratio was similar to 201Tl defect count ratio on the initial rest 201Tl scan (0.57 +/- 0.03 versus 0.56 +/- 0.02, p = ns), but significantly less than 201Tl defect count ratio at 2 hr (0.57 +/- 0.03 versus 0.65 +/- 0.02, p < 0.05). CONCLUSION: In a low-flow model with profound systolic dysfunction, myocardial 99mTc-tetrofosmin uptake ( > 50%) reflective of viability was observed in the asynergic zone perfused by the stenotic LAD. PMID- 8708785 TI - Evaluation of human transferrin radiolabeled with N-succinimidyl 4-[fluorine 18](fluoromethyl) benzoate. AB - Iron metabolism plays a key role in cell proliferation and survival in rapidly growing cancer cells. Uptake is mediated by the carrier protein transferrin. The increased need for iron has been used as a method to target tumors and there is well-documented evidence that certain tumors can be imaged with tracers such as 67Ga, that mimic transferrin-mediated iron uptake. To obtain a tracer that would be better able to quantitate transferrin kinetics and indirectly evaluate iron metabolism, we have labeled human transferrin with the positron emitter, 18F, with a one-step high-specific activity method developed in our laboratory. METHODS: We measured the binding affinities of [18F]diferric (holo-) and iron free (apo-) transferrin on two human cell lines. We also compared cellular uptake of [18F]holo-transferrin and [67Ga]citrate in various conditions, and washout of label incorporated into cells. RESULTS: The binding affinity of [18F]holo transferrin was found to be the same as that reported for [125I]holo-transferrin. In our hands there was no significant difference in binding affinity between diferric holo-transferrin and iron-free apo-transferrin. [18F]holo-transferrin uptake rapidly reaches a steady-state equilibrium between the intracellular and extracellular environment, while gallium accumulation linearly increases with time. [18F]holo-transferrin is rapidly recycled out of the cell with similar kinetics to those reported for [125I]holo-transferrin. CONCLUSION: [18F]holo transferrin displays the properties of native transferrin and appears suitable for quantitative evaluation of transferrin kinetics in vivo. PMID- 8708784 TI - Evaluation of myocardial viability with iodine-123-BMIPP in a canine model. AB - The tracer 123I-BMIPP was examined for its ability to reflect myocardial lipid metabolism. Studies in mice indicate that myocardial BMIPP uptake correlates with ATP content. Details, however, of myocardial accumulation in the ischemic period with either infarct or ischemia are not well documented. METHODS: Sixteen adult mongrel dogs were investigated. The occluded left anterior descending artery (LAD) alone was reperfused to make the ischemic area, and the first diagonal branch of the LAD was kept occluded to make the infarct area. Regional wall motion was evaluated by echocardiography in the short-axial view from the epicardium. Tissue blood flow was calculated using nonradioactive colored microspheres. Changes in blood glucose levels, lipid levels and lactate extraction were examined in blood collected from the aorta and great cardiac vein (GCV). The ATP concentration and BMIPP count were determined by high-performance liquid chromatography and gamma-counter, respectively. RESULTS: Two hours after reperfusion, blood flow decreased to 20% +/- 5% in the infarct area and 64% +/- 9% in the ischemic area (p < 0.05), despite comparable wall-motion reduction (32% +/- 5% and 42% +/- 12% in the infarct and ischemic areas, respectively). BMIPP content and ATP concentration showed parallel reduction: 40% +/- 7% and 75% +/- 4% (p < 0.05) of BMIPP and 32% +/- 9% and 69% +/- 7% (p < 0.05) of ATP in the infarct and ischemic areas, respectively. The nonesterified fatty acid extraction, defined as flow x ([artery] - [GCV]), decreased to 87% +/- 5.6% during occlusion and 75% +/- 20.1% 2 hr after reperfusion, as compared with the control value. CONCLUSION: BMIPP uptake correlated well with lipid metabolism and tissue ATP levels and may prove useful in differentiating myocardial infarction from ischemia in the acute phase of ischemic episodes. PMID- 8708786 TI - Oxido-reductive state: the major determinant for cellular retention of technetium 99m-HMPAO. AB - Several clinical observations have suggested that HMPAO cerebral uptake might be related not only to regional cerebral perfusion but also to the nature of the lesion. Our aim was to investigate at the cellular level the nature of the process(es) involved in HMPAO accumulation in vitro. METHODS: Time-course incorporation of HM-PAO was studied in a fast-growing human premonocytic line, U937, in a human astrocytic-derived cell line, U373 and a human hybridized endothelial cell line, EaHy926. Minimal differences of HMPAO retention between these cell lines were observed and plateau of %U(HMPAO) (cpm cells/cpm standard of injected) were achieved within 2 hr. Because HMPAO cell retention was related to the intracellular content in glutathione, experiments studying effects of redox were conducted by preexposing U937 cells to D, L dithiothreitol or 2 Mercaptoethanol. RESULTS: Overnight incubation with NAC or BSO did not significantly modified the kinetic of 99mTc-HMPAO incorporation while overnight incubation with NAC resulted in a 2-fold increase in intracellular glutathione content and overnight incubation with BSO nearly abolished the intracellular glutathione content. At the opposite, presence of these reducing agents in the medium during the experiments completely abolished 99mTc-HMPAO retention. CONCLUSION: Our data thus provide in vitro evidence to support that overall intracellular retention of HMPAO is more dependent upon the redox activity of the tissue than the intracellular glutathione content. SPECT-HMPAO may accurately reflect regional cerebral blood flow in a normal state but possibly not in all pathological situations in which cell metabolism disturbances are characterized by alterations in the redox status. PMID- 8708787 TI - Collimator design for improved spatial resolution in SPECT and planar scintigraphy. AB - This paper evaluates the design of a new planar-concave collimator with nonuniform response that better matches the body shape than conventional collimators. METHODS: The collimator properties are evaluated and assessed by means of both a stimulation program and an experimental test using a prototype planar-concave collimator. RESULTS: The results, for points located 150 mm from the axis of rotation, demonstrate that the ratio of radial and tangential spatial resolution in SPECT with the new collimator decreased by 40%, as compared to SPECT with a standard collimator. In planar scintigraphy, the spatial resolution improved correspondingly from 10.9 mm (FWHM) to 7.8 mm in lateral areas of the body. CONCLUSION: The new collimator reduces nonisotropic blurring in SPECT in addition to improving spatial resolution in both planar scintigraphy and in SPECT. PMID- 8708788 TI - Comparative methods for quantifying thyroid volume using planar imaging and SPECT. AB - SPECT enables improved accuracy over planar imaging in the determination of the volume since it is derived from three-dimensional data rather than from a two dimensional projection with a certain geometric assumption regarding the thyroid configuration. METHODS: By using the phantoms of known volume, it was possible to estimate the accuracy of three different methods of determining thyroid volume from planar imaging used in clinical routine: the standard method used at Malmo General Hospital; a modified version of this standard method; and the method used in Lund Hospital in combination with different ways of defining the regions of interest (ROIs), and to assess the accuracy of the adaptive threshold of gray level histogram method based on SPECT imaging which determines a threshold that maximizes the separability of two classes (object and surround). RESULTS: The correlation coefficient (r) and the regression equation between the true (x) and the calculated volume (v) were as follows: r = 0.99 and y = 0.98x + 3.6 using SPECT and the gray level histogram method for edge detection combined with attenuation and scatter corrections, while r = .97 and y = 0.67x + 3.2 using the standard method based on planar scintigraphy. The standard method as used in routine was found to produce large errors (24.8%). The error on the volume estimate was reduced to approximately 7% for volumes in the range 16-to-75 ml using SPECT. CONCLUSION: Compared with conventional scintigraphy, thyroid phantom volumes were most accurately determined with SPECT when attenuation and scatter corrections are performed, which allows accurate radiation dosimetry in humans without the need for assumptions on organ size or concentration. PMID- 8708789 TI - "Father of emission tomography" receives 1996 Cassen Prize. PMID- 8708790 TI - Cultural cohesion versus collision. A model for facilitating organizational mergers. AB - Healthcare delivery systems are experiencing sweeping changes; acquisition and merger activities abound. These activities have the potential to greatly impact employees and their quality of work-life. The authors describe how one nursing organization in a large metropolitan teaching hospital successfully merged two separate nursing departments by dealing with the essence of these organizations' cultures. A conceptual model was developed that will facilitate future organizational field research in the areas of corporate culture and change process. PMID- 8708791 TI - The intrinsically motivated nurse. Help and hindrance from evaluation feedback sessions. AB - Self-determination theory describes the working conditions necessary for the experience of self-motivation. Performance evaluation sessions represent extraordinary opportunities to enhance motivation, yet often are approached with anxiety. An innate desire for feedback meets with a learned response of fear, by both the evaluator and evaluatee. Psychological fusion theory explains the source of this conflict and what nurse executives can do to use these sessions for motivational impact while ameliorating the attendant stress. PMID- 8708792 TI - Partners in distance learning. Project outreach. AB - Distance learning refers to any educational experience in which the instructor (teacher) is separated from the student (learner) by geographic distance. Partnerships are being established between institutions of higher education and healthcare organizations to achieve a mutual goal, that of educating employees to work in a rapidly changing workplace environment. The authors describe one such partnership and the common issues confronted by both the academic and service institutions in implementing an outreach education program. The authors propose that these partnerships can be effective in implementing distance learning programs that meet the ongoing educational needs of nurses living and working in rural and underserved environments. PMID- 8708793 TI - Cost analysis of intermediate care versus intensive care for the neurosurgery patient. PMID- 8708794 TI - Employment discrimination experiences of registered nurses. AB - Successful recruitment, retention, motivation, and satisfaction of nurses should be of primary concern to all employer organizations and supervisors. Employment discrimination is an onerous stress that negatively interferes with success. The authors discuss the legal concepts of employment discrimination, report the discrimination experiences of the nurses in their research project, and make recommendations for confronting and preventing employment discrimination. PMID- 8708795 TI - Emerging knowledge. AB - The healthcare delivery system is undergoing great upheaval as individual providers respond to market pressures for aggressive cost reduction. Roles, relationships, processes, partnerships, and financing arrangements are on the table in the great race for competitive survival. The IDN department addresses issues inherent in the evolution of free-standing acute care centers and private medical staffs into integrated delivery networks. It globally explores alternative managed care and direct contracting at-risk strategies. PMID- 8708796 TI - Community development theory. Planning a community nursing center. AB - The authors describe the use of community development theory to assess the need for a community-based, nurse-managed primary care clinic. A community development model provided the framework for citizen participation in identifying collective health needs of public housing residents. The model facilitated the following: 1) planning for delivery of culturally appropriate primary care services that respond to health needs perceived by community residents; 2) ensuring acceptability and use of services; and 3) empowering residents to take responsibility for their own health. This article focuses on the assessment phase of the model and meeting the perceived needs of community residents. PMID- 8708797 TI - Improving healthcare quality. Application of the Baldrige process. AB - The Malcolm Baldrige National Quality Award was established in 1987. Although primarily an award for business and industry, pilot criteria for healthcare settings have been developed. These criteria can be used by healthcare organizations to conduct internal evaluations of their institutions. Benefits of such an evaluation include improvement of an organization's effectiveness and improved healthcare quality for stakeholders. PMID- 8708798 TI - Reflecting professional values in the philosophy of nursing. AB - If nurses are to remain central in restructuring efforts, their professional values must be at the core. Nurse administrators play a key role in promulgating values. The authors examined philosophy of nursing documents to identify professional values and found them to be lacking. They discuss their findings and their implications for nurse executives. PMID- 8708799 TI - JONA's Directory of Consultants to Patient Care Administration. PMID- 8708800 TI - Nurse administrator employment in group practices. AB - The decline in managerial positions in hospitals has resulted in nurses' increased interest in management opportunities in other healthcare system sectors. This study reports the responses of 963 medical group practices to a survey of educational preferences and support mechanisms for the preparation of administrators for this type of setting. The findings raise issues for nurse managers seeking nonhospital employment and for nursing schools that have defined their educational mission as encompassing the preparation of nurse managers. PMID- 8708801 TI - Nursing in Brazil. AB - The goal of International Perspectives is to share the views of nurse leaders, from countries other than the United States, on global healthcare issues. The focus is on identifying healthcare management problems and their implications, research priorities, policy development, and ethical-moral dilemmas faced by nurse administrators. You are encouraged to send the names and addresses of international nurse leaders who might be interested in participating, as well as your ideas, for format and content of the department. PMID- 8708802 TI - Defining terminology for improved breastfeeding research. PMID- 8708803 TI - Perimenopausal and postmenopausal hormone replacement therapy. Part 2. Hormonal regimens and complementary and alternative therapies. AB - As midwives increasingly provide care to women throughout the lifecycle, they need to become familiar with a variety of treatments for perimenopausal and postmenopausal problems and preventive measures for the potentially disabling conditions that can accompany aging. This article reviews various regimens of hormone replacement therapy. It discusses types of estrogen and progestin, appropriate dosages, routes of administration, and duration of therapy. It offers guidelines for initiation, contraindications, and required follow-up. It addresses the management of side effects and problems. The article briefly reviews a variety of complementary and alternative therapies including various self-help measures, such as nutrition and exercise, and selected herbal treatments. Part 1 of this article, which appeared in the January/February 1996 issue of JNM, reviewed the literature on recommended therapeutic and preventive indications for hormone replacement therapy, providing guidelines for the provision of informed consent. PMID- 8708804 TI - Use of the Cytobrush for Papanicolaou smear screens in pregnant women. AB - A prospective, randomized clinical trial was conducted to study the safety and efficacy of the Cytobrush for Papanicolaou smear screens in 252 pregnant women. Two groups-Ayre spatula plus Cytobrush and Ayre spatula plus cotton swab-were compared. There was a higher incidence of spotting with each of the three devices as compared with studies of nonpregnant women. The highest incidence of spotting at time of exam occurred with the Cytobrush; however, this spotting was not clinically significant. Eleven first-trimester women experienced a pregnancy loss; five of these losses were due to a hydatidiform mole or missed abortion that occurred before the exam. Six women (2%) aborted spontaneously compared with a 10% incidence reported in the literature. It was concluded that all three devices are safe for use in pregnancy. In addition, the most adequate screen for retrieval of squamous epithelial cells was found to be the Ayre spatula, while the Cytobrush proved most effective for retrieval of endocervical epithelial cells. PMID- 8708805 TI - Current assessment of the use of intrauterine devices. AB - The need for effective, long-term contraception remains a significant issue. Intrauterine devices (IUDs) have evolved to the point where currently available devices are comparable, in safety and efficacy, to oral contraceptives. Due to concerns regarding previous devices, available IUDs remain largely underused by midwives and all other providers of reproductive health care. Recent medical data refute many of the medical-legal misconceptions. Currently available IUDs are safe and effective and represent a suitable contraceptive alternative for the appropriate patient. PMID- 8708806 TI - The home study program on home birth. PMID- 8708807 TI - The home study program on home birth. PMID- 8708808 TI - The home study program on home birth. PMID- 8708809 TI - Perimenopausal and postmenopausal hormone replacement therapy: Part 1. An update of the literature on benefits and risks. PMID- 8708810 TI - Findings of the American College of Nurse-Midwives Annual Membership Surveys, 1993 and 1994. AB - The findings of the 1993 and 1994 American College of Nurse-Midwives Membership Surveys are presented and compared with a 1991 survey. The approximately 3,600 respondents had a mean age of 43 years, and most members were female and Caucasian. Two-thirds of the certified nurse-midwives (CNMs) who responded had earned master's degrees. Approximately one-half of the respondents were employed by physicians or hospitals, and 71% reported that their primary employment involved clinical care with attendance at births. More than 90% of CNM respondents who reported site of birth attended births in hospitals. There has been a demonstrated increase in racial/ ethnic diversity of the membership since the 1991 survey. PMID- 8708811 TI - Development of an academic nurse-midwifery service program. A partnership model between medicine and midwifery. AB - Academic nurse-midwifery services have shown themselves to be strong collaborators with medical education. The development and functioning of an academic nurse-midwifery service program built on a partnership model between medicine and midwifery are highlighted. Organizational relationships, philosophical approach, and practice dimensions including responsibilities for clinical practice, medical student, and obstetrical resident education are explored. As the obstetrical patient pool diminishes in academic service settings, this model may prove useful. PMID- 8708812 TI - Practice styles. A comparison of obstetricians and nurse-midwives. AB - Iain Chalmers and the National Perinatal Epidemiology Unit, Oxford, England, developed a comprehensive listing of perinatal care procedures shown to reduce the frequency of adverse outcomes during pregnancy and childbirth. This list was used as a framework for a pilot study conducted in 1992 that reviewed similarities and differences in opinion and practice style between certified nurse-midwives (CNMs) and obstetrician/gynecologists. Twenty CNMs and 57 obstetrician/gynecologists who were active clinical practitioners in San Diego commented on 24 items drawn from Chalmers' work. The CNMs were more likely to favor the availability of social and psychological support variables and to use them in their practice. The groups were more alike than different in their views concerning preventive interventions during the prenatal period. CNMs were more likely to support the availability of alternatives to maternal positions for labor and birth, exhalatory breathing, and delayed pushing and less likely to support the availability of electronic fetal monitoring, epidural anesthesia, episiotomy, and active management of the third stage. A small sample size and limited response rate restricted interpretation and generalizability of these data. Nevertheless the data offer support for other studies with similar findings. They also suggest that health system administrators should inform women and families about differences in practice styles before families select from among the various insurance options that may, in the end, restrict the choice of provider or birth setting. PMID- 8708813 TI - The underuse of women's health care services in hungary. A preliminary study. AB - In order to collect baseline data for a 2-year Women's Health Program in Hungary, a survey was conducted of 369 women residing in one community setting. The purpose of this descriptive exploratory study was to gather preliminary data that would help identify perceptions of inhibitors to utilization of women's health services. An additional purpose was to identify perceived health needs in the study setting and to make recommendations for changes in the existing services. The survey results indicated that women were satisfied overall with their health care. Equivocal responses suggested, however, that patient-provider interactions and lack of understanding regarding preventive health care may contribute to the underuse of services. The authors suggest that public feedback that includes women's opinions and perceived needs is essential in health program planning and evaluation. In order to accomplish this, standardized Hungary-specific research instruments must be developed. PMID- 8708814 TI - Examining separated symphysis pubis. AB - Separated symphysis pubis (SSP) is a rare condition that results in a separation of the symphysis pubis bone in late pregnancy or during delivery and occurs in otherwise healthy pregnancies as a result of hormonal and/or biomechanical factors. Several researchers have examined the issue in the Scandinavian countries, where it appears to be more prevalent possibly due to a genetic link. Symptoms range from mild discomfort to total debilitation. Differential diagnosis includes exclusion of more serious medical conditions. The nurse midwife's role is prompt diagnosis, medical consultation, support, and education. PMID- 8708815 TI - Rural midwifery and electronic communications. AB - Electronic communications can introduce a whole new world of friends, knowledge, and opportunity to the rural midwife. It can decrease isolation and improve writing skills. There are opportunities for midwives with computer skills to take part in research and participate on electronic bulletin boards designed specifically for midwives. It is the author's opinion that more midwives, especially rural midwives, should avail themselves of this knowledge. PMID- 8708816 TI - Decrease of serum ascorbic acid concentrations in patients with diabetic macroangiopathy. AB - The relationship between serum ascorbic acid (AA) and diabetic macroangiopathy was studied. Fifty-six patients with noninsulin-dependent diabetes mellitus were examined, together with 20 healthy controls matched for age against the diabetes patients. Aortic pulse wave velocity (PWV) was taken as an index of the severity of atherosclerosis. The level of serum AA in diabetic patients was significantly lower than that of the controls. Among the diabetic groups, those with elevated PWV levels by age demonstrated a significant drop in AA. No significant differences were seen in the level of serum dehydroascorbic acid (DHAA) between patients and controls, nor were there any significant differences among patient groups. The concentration of serum AA was inversely related to the risk factors of atherosclerosis, such as body mass index, Apo B/ Apo A-I ratio, thiobarbituric acid-reactive substances (TBARS), and microalbumin in urine. It was inferred from these findings that the depletion of serum AA was apparent in diabetics with advanced atherosclerosis. PMID- 8708818 TI - Comparison of young women's salt intake between Japan and England: consideration of different measurement methods in national nutrition surveys. AB - Japanese salt intake is calculated through the Food Composition Table in the National Nutrition Survey, whereas English salt intake is through urinary sodium excretion in 24 h. This study is a comparison of salt intake between Japan and England. As different measuring methods were used in both National Nutrition Surveys, we recalculated using the same measuring method. Seven Japanese young women's sodium intakes were calculated through the Food Composition Table, and the amounts of their urinary-excreted sodium were measured. These quantities were compared, and a coefficient (87.7%) was obtained. The Japanese young women's salt intake was 9.74 g/day from the Japanese National Nutrition Survey, and the coefficient was multiplied by 8.51 g/day. While comparing these experimental results with English data in 1992, the English salt intake converted from sodium intake of 3,040 mg/day was 7.72 g, 0.79g/day lower than the Japanese salt intake. PMID- 8708817 TI - Age-related changes of alpha-tocopherol transfer protein expression in rat liver. AB - Developmental changes in expression of alpha-tocopherol transfer protein (alpha TTP) after birth were investigated using rats with respect to plasma changes of tocopherols. The expression of alpha-TTP in the neonatal rat liver, which was very low immediately after birth, increased steadily during the two weeks of life before weanling and reached the adult level at four weeks. During the suckling period, the plasma ratio of alpha-tocopherol to gamma-tocopherol linearly increased, because plasma alpha-tocopherol which was low immediately after birth, increased rapidly during the period, while gamma-tocopherol remained unchanged. The increase in the ratio seemed to correlate with the developmental expression of alpha-TTP in the liver during this period. The ratio also reached the adult level after four weeks. The expression of alpha-TTP was investigated using primary cultured rat hepatocytes. The expression of alpha-TTP was found to be extremely low after 20h of culture. The decrease in alpha-TTP expression was exacerbated by adding epidermal growth factor to the culture medium and was inhibited by adding dexamethasone. These observations suggest that expression of alpha-TTP may be affected by the state of hepatic differentiation. PMID- 8708819 TI - Pattern of fecal endogenous nitrogen excretion in rats fed leguminous diets. AB - The main objective of this investigation was to establish the pattern in relation to time of the rat fecal endogenous nitrogen excretion by continuous feeding of balanced diets containing common peas, cowpeas or common beans as the protein sources (10% protein), labeled with 1.000 atoms % of 15N-excess. Nitrogen of endogenous origin was measured by the isotope dilution method in a 6-day experiment. Fecal excretion of endogenous nitrogen of rats fed the leguminous diets was roughly twice that of rats fed the non-protein diet (88 mg, 42 mg), and the excretion of total fecal nitrogen did not differ among leguminous diets. From the third to the sixth day of the experiment, the endogenous nitrogen excretion, either as a percentage of quantity (mg), attained a statistically non different value (p > 0.05). A common pattern of excretion of fecal endogenous nitrogen as a function of time was expressed by a strong negative (r < -0.95) power regression (y = A.x-B) for the common pea, the cowpea or the common bean diets. Conversely, the excretion of dietary nitrogen did not show a common pattern as a function of time for all experimental diets. PMID- 8708820 TI - High phosphatidylcholine hydroperoxide level in plasma of guinea pigs with low and excess supplementation of ascorbic acid. AB - Graded amounts (0, 50, 500 and 5,000 mg/liter) of ascorbic acid (AsA) were given in drinking water to guinea pigs for 21 days to prepare AsA-deficient, low-AsA, moderate-AsA and excess-AsA animals, and the plasma phospholipid hydroperoxide level and lipid concentration were quantitatively determined to investigate the antioxidant effect of AsA in vivo. Phosphatidylcholine hydroperoxide (PCOOH) was a predominant phospholipid hydroperoxide present in the plasma, and the PCOOH concentration was significantly higher in AsA-deficient, low-AsA and excess-AsA animals (80.4 nM, 54.8 nM and 42.2 nM, respectively) as compared with that in moderate-AsA animals (27.2 nM). Hyperlipidemic plasma characterized as high cholesterol and high triacylglycerol concentrations was confirmed in AsA deficient animals. Molar ratios of plasma AsA and alpha--tocopherol against 10(4) moles of phospholipids were significantly lower in AsA-deficient and low-AsA animals (0.6-2.1 and 5.5-8.5, respectively) than in moderate-AsA and excess-AsA animals (14.2-18.0 and 11.2-11.9, respectively). In plasma, a high correlation coefficient (r = 0.979) was observed between PCOOH and AsA for which there was optimum AsA level to keep the low PCOOH and such correlation was stronger than that (r = 0.558) observed with alpha-tocopherol. The results indicated that AsA has an important function to control the phospholipid hydroperoxide level in plasma and that moderate supplementation of AsA is required to reveal its optimal antioxidant effect in vivo. The present study also showed that AsA-deficiency especially invites an increase in plasma PCOOH together with a hyperlipidemic state which are risk factors in developing atherogenesis. PMID- 8708821 TI - Pharmacological effects of phosphatidylserine enzymatically synthesized from soybean lecithin on brain functions in rodents. AB - Soybean transphosphatidylated phosphatidylserine (SB-tPS) was prepared from soybean phosphatidylcholine by transphosphatidylation using phospholipase D, and the fatty acids composition and pharmacological properties were compared with those of bovine brain cortex-derived phosphatidylserine (BC-PS) which was reported to improve cognitive disorders of senile dementia patients by oral administration (300 mg/day). The molecular species of SB-tPS are rich in linoleic and palmitic acids whereas those of BC-PS are stearic and oleic acids. Despite the differences in fatty acid composition, SB-tPS displayed significant activities on the increase in brain glucose concentrations in mice (79 mg/kg, i.v.) and the restoration of scopolamine-induced amnesia in rats (60 mg/kg, i.p.) as did BC-PS. These results suggest the possibility that SB-tPS may prevent and/or improve senile dementia by oral administration. PMID- 8708822 TI - Ovariectomy decreases osteogenetic activity in rat bone. AB - The purpose of this study was to investigate the effect of ovariectomy on osteoinductive activity in the bone in rat. Homograft implantation of decalcified humeral diaphysis from ovariectomized or sham-operated rats was performed and harvested after several time periods. A significant decrease in bone induction was found in terms of soft X-ray photography, alkaline phosphatase activity, mineral content and expression of osteocalcin (BGP; bone gla-protein) in the implants from the ovariectomized group in comparison to those from the sham operated animals. This result suggested that the level of osteoinductive activity, probably due to bone morphogenetic protein, decreased in ovariectomized animals. PMID- 8708824 TI - Effective use of the research consultant. AB - A principal investigator (PI) often includes a consultant in a proposal for a research project either at the request of the funding agency or because a consultant may add expertise to the research team. The most effective use of the consultant may vary according to the needs and gaps in areas of expertise of the team. Lippitt and Lippitt (1978) developed a model for guiding consultation that has been adapted in this article to guide not only the consultant but also the researcher in how to effectively use a consultant. The model is adaptable whether the principal investigator is new to designing and implementing research projects and needs a consultant expert in the research process or the PI has much research experience and needs a clinically expert consultant. The six phases of this model from contact and entry through contract completion is a step-by-step guide that is adaptable to the needs of the PI and research team. PMID- 8708823 TI - The effects of isomaltulose-based oligomers feeding and calcium deficiency on mineral retention in rats. AB - We examined the effects of isomaltulose-based oligomers (IBOs) on the mineral content of the whole blood, kidney, liver and tibia in calcium deficient and calcium sufficient rats. Twenty-eight Wistar rats were divided equally into 4 groups and fed with the following diets ad libitum for 4 weeks: (1) calcium sufficient diet (Ca+, IBOs-), (2) calcium-sufficient-IBOs supply diet (Ca+, IBOs+), (3) calcium-deficient diet (Ca-, IBOs-), (4) calcium-deficient-IBOs supply diet (Ca-, IBOs+). There were no significant differences in final body weights among the groups. Food consumption in the calcium-deficient groups was higher than that in the calcium-sufficient groups. The tibia weight was significantly decreased, and the calcium, magnesium and phosphorous contents significantly decreased, and iron content was significantly increased in the tibia of calcium-deficient rats. On the other hand, in IBOs feeding rats, tibia weight, and calcium, magnesium and phosphorous contents were significantly increased, and iron content was significantly decreased. These findings suggest that IBOs feeding improves mineral retention especially in a state of calcium deficiency. PMID- 8708825 TI - Bibliography: regulatory issues. PMID- 8708826 TI - The New Health Professionals. PMID- 8708827 TI - From RN to BS: one program's experience with the curriculum revolution. AB - An upper-division nursing program's framework for socialization of the registered nurse to the professional nursing role is presented. Critical thinking is linked to the transformation process, wherein the learner moves from proficiency in technical knowledge to experience that is analyzed and organized by critical thought. The impact on teaching-learning approaches, experience with the Watson Glaser Critical Thinking Appraisal, and institution of a portfolio system designed to track students' perceptions of changes in critical thinking are discussed. PMID- 8708828 TI - Columbia-Presbyterian nursing alumni: a case study in American social history. AB - Using survey data obtained from 2,601 undergraduate nursing alumni, this descriptive study demonstrates the ways nursing's development has reflected America's social history. PMID- 8708829 TI - Regaining self-esteem: strategies and interventions for the infertile woman. AB - Infertility can cause women to lose their sense of power and control, thus diminishing their self-esteem. Terminology, technology, and testing may contribute to this sense of failure. Nurses can intervene by recognizing this negative consequence of infertility and using strategies within the nursing model. PMID- 8708830 TI - Understanding the Apgar score. AB - Apgar scores are determined for every neonate born in a U.S. hospital. Despite the frequency with which the scores are calculated, they are not always accurate. In addition, some individuals attempt to use the scores to substantiate certain claims, such as birth asphyxia. This article discusses some of the common misunderstandings and limitations of the Apgar score and suggests measures for improvement. PMID- 8708831 TI - Management of the obstetric patient with tuberculosis. AB - Tuberculosis in obstetric patients presents a challenging nursing problem to facilities caring for populations at high risk: the economically disadvantaged, minorities, the foreign-born, and patients with human immunodeficiency virus. Coordination of public health, outpatient, and inpatient services is essential for obtaining the desired outcome of a healthy family unit with a minimum of interruption of the attachment process. This article presents issues related to nursing care and medical management of tuberculosis in the perinatal period. PMID- 8708832 TI - Physical abuse, smoking, and substance use during pregnancy: prevalence, interrelationships, and effects on birth weight. AB - OBJECTIVE: To establish the singular and combined occurrence of physical abuse, smoking, and substance use (i.e., alcohol and illicit drugs) during pregnancy and its effect on birth weight. DESIGN: Prospective cohort analysis. SETTING: Urban public prenatal clinics. PARTICIPANTS: 414 African American, 412 Hispanic, and 377 white pregnant women. MAIN OUTCOME MEASURE(S): Occurrence of physical abuse was 16%; smoking, 29.5%; and alcohol/illicit drug use, 11.9%. Significant relationships existed between physical abuse and smoking for African American and white women. For African American women, 33.7% of women who were not abused smoked, versus 49.5% of women who were abused (chi 2 = 8.21; df = 1; p < 0.005). Alcohol/illicit drug use was 20.8% for nonabused women compared with 42.1% for abused women (chi 2 = 18.18; df = 1; p < 0.001). For white women, 46.6% of women who were not abused smoked, versus 59.6% of those who were abused (chi 2 = 5.22; df = 1; p < 0.005). As a triad, physical abuse, smoking, and alcohol/ illicit drug use were significantly related to birth weight (F[3, 1040] = 30.19, p < 0.001). CONCLUSIONS: Physical abuse during pregnancy is common, readily detected with a five-question screen, and associated with significantly higher use of tobacco, alcohol, and illicit drugs. Clinical protocols that integrate assessment and intervention for physical abuse, smoking, and substance use are essential for preventing further abuse and improving smoking and substance cessation rates. PMID- 8708833 TI - The effects of childhood sexual abuse on minority adolescent mothers. AB - OBJECTIVE: To examine the incidence and effects of childhood sexual abuse experiences on the lives of adolescent mothers who were Mexican American and African American. Specifically, the personality characteristics of the adolescent mothers who experienced sexual abuse as assessed using the Millon Adolescent Personality Inventory were compared with those of minority adolescent mothers who were not abused sexually. DESIGN: An exploratory descriptive design. SETTING: Four public agencies caring for adolescent mothers in two urban centers in Texas. PARTICIPANTS: Mexican American and African American adolescent mothers (N = 124), ages 13-20 years. MAIN OUTCOME MEASURES: The Millon Personality Inventory, the Life Event Scale-Adolescents, the Childhood Sexual Experience Survey, and a demographic data sheet. RESULTS: Forty-four percent (n = 54) of the respondents reported childhood sexual abuse experience. Data from 111 subjects (two groups, abuse and nonabuse) were analyzed by MANCOVA techniques. Significant differences were found between the two groups when the effect of the Life Event Scale Adolescents was controlled (Multivariate F[12, 97] = 3.055, p = 0.001). Univariate F tests (df1, 108) showed significant differences in self-concept (F = 18.434, p < 0.000), personal esteem (F = 8.819, p = 0.004), body comfort (F = 10.442, p = 0.002), sexual acceptance (F = 9.918, p = 0.002), peer security (F = 10.412, p = 0.002), family rapport (F = 9.801, p = 0.002), academic confidence (F = 5.764, p = 0.018), social conformity (F = 4.552, p = 0.035), scholastic achievement (F = 19.991, p < 0.000), and attendance consistency (F = 10.009, p = 0.002). The mothers in the abuse group had consistently poorer scores than those in the nonabuse group. CONCLUSIONS: These findings suggest that childhood sexual abuse may be related to vulnerability for adolescent motherhood. PMID- 8708834 TI - Family centered intrapartum care: revisiting an old concept. AB - The historic development of family centered care to the present time is traced, and future development is proposed. A family systems approach can help nurses focus on the family as a unit and on individual members. This requires nurses to develop skills in working with multiple family members, provide care and advocacy for varying and sometimes conflicting family needs, and promote family clarification of their own values and priorities. Such an approach can help intrapartum families add to their experience of coping with stress in a way that builds family cohesion and mutual support. PMID- 8708835 TI - Parturition and fetal adaptation. AB - The fetus as "patient" during labor and birth has become an increasingly important concept during the past 20 years. However, what is understood about fetal status during labor and how the fetus prepares for its approaching separation from its mother? Current information indicates that not only is the term fetus well prepared for the adaptation to extrauterine life, but this transition is facilitated by normal labor. PMID- 8708837 TI - American Association of Oral and Maxillofacial Surgeons 78th annual meeting and scientific sessions. Miami Beach, Florida, September 18-22, 1996. Abstracts. PMID- 8708836 TI - Epidurals redefined in analgesia and anesthesia: a distinction with a difference. AB - Advances in our understanding of pain have created a new and meaningful distinction between epidural analgesia and anesthesia. The principles underlying this important difference are reviewed from a childbirth perspective and promote a reconsideration of the role of pain relief in labor versus the need for anesthesia at delivery. An epidural analgesic technique is presented with fresh implications for several old intrapartum practices. The analgesia/ anesthesia distinction in epidural therapy offers new opportunities and flexibility for safety and fulfillment in childbirth. PMID- 8708838 TI - Vinorelbine tartrate: a promising new chemotherapeutic agent. AB - Vinorelbine tartrate (Navelbine, Burroughs Wellcome Company, Research Park, NC) is a semisynthetic analog of vinblastine that has been approved by the Food and Drug Administration for use in the treatment of advanced, unresectable non-small cell lung cancer. It has a favorable safety profile and can safely and easily be administered in the outpatient setting on a weekly basis. The major toxicity is severe, granulocytopenia (absolute neutrophil count < 500 cells/mm3), that occurs in approximately 40% of treated patients. This granulocytopenia is rapidly reversible, and does not increase in severity with subsequent cycles. Despite the high incidence of granulocytopenia, only about 8% of patients require hospitalization for the treatment of febrile neutropenia. This article also reviews information concerning the incidence and management of the other major side effects including constipation, paresthesias, decreased deep tendon reflexes, myalgia, and injection site reactions. Guidelines for the safe administration of this agent along with suggestions for patient education are also presented. The potential significance of the impact that this new agent may have in the treatment of advanced, unresectable non-small cell lung cancer and advanced breast cancer is stressed. PMID- 8708839 TI - Comparison of two percutaneous intravenous "midline" catheters in cystic fibrosis. AB - Administration of intravenous antibiotics in cystic fibrosis has been facilitated by the use of midline catheters; percutaneous lines inserted through a peripheral vein and advanced into a large but noncentral vein. In a randomized study, a 23 gauge silastic catheter (Vygon EC, Cirencester, United Kingdom) was compared with the Hydrocath (Viggo-Spectromed, Swindon, United Kingdom), a 22-gauge hydrophillic coated polyurethane catheter inserted using the Seldinger technique. Fifty eight courses of intravenous antibiotics were given, 28 through the Hydrocath (median age 11 years, range 1.5-17.5 years) and 30 through the silastic catheter, (median age 11 years, range 0.5-17.5). Mean line survival was equal. The Hydrocath took longer to insert and was associated with more pain on insertion. However, administration of antibiotics was easier through the Hydrocath and overall satisfaction was higher in those who had the Hydrocath. Both catheters performed well, but administration of antibiotics was easier through the Hydrocath. PMID- 8708840 TI - Peripherally inserted central catheter insertion problems associated with topical anesthesia. AB - Application of the topical anesthetic cream to the antecubital fossa 60 minutes before insertion of the peripherally inserted central catheters resulted in significant vasoconstriction and/or vasospasm in 24 patients. Catheter insertion was difficult in all 24 patients. Advancement of the peripherally inserted central catheter was impossible in six patients. Of the 51 patients that did not have the topical anesthetic pre-catheter insertion, there were no difficulties in placing the lines, nor were there any vasoconstriction and/or vasospasm. This group included 66 adults and 9 children. PMID- 8708841 TI - Intravenous nursing services: strategies for success. AB - The intent of this article is to provide intravenous nurses with options for marketing and promoting their IV teams in institutions to enhance viability of the team concept and promote quality nursing care for the consumer. The article supplies options for a business plan to present to administration to promote the team concept both in the hospital and in alternate site settings. PMID- 8708842 TI - Managing change while maintaining quality in home infusion therapy. AB - With the multiple external forces dictating rule and regulatory, third party payer/reimbursement, and accreditation changes for the home infusion industry, identifying methods for managing the changes while maintaining quality in patient and organization outcomes has become essential. The challenge for the home infusion manager, nurse, pharmacist, and other healthcare professionals is to effectively anticipate, measure, monitor, and manage changes for the operations and clinical aspects of service. PMID- 8708843 TI - Hyperemesis gravidarum: implications for home care and infusion therapies. AB - Nausea and vomiting afflicts up to 80% of pregnancies, with the more severe form, hyperemesis, complicating about 1%. Hyperemesis in pregnancy causes fluid volume deficit, starvational ketoacidosis, and at times, metabolic alkalosis with hypokalemia. Significant weight loss may occur and reflect fluid as well as lean tissue loss. The cause is unknown, and although prognosis is generally good with aggressive fluid replacement and nutritional support, there is potential for decreased birth weight. Management strategies for hyperemesis gravidarum, including hydration, enteral and parenteral nutrition, antiemetic therapies, monitoring needs, and psychosocial concerns are discussed with applications made to the home care setting. PMID- 8708844 TI - The history of intravenous therapy. AB - Modern IV therapy is less than a century old. Yet, it was known that medications could be injected into a vein as early as the 1600s. Because of a lack of scientific methods, original attempts to deliver IV fluids and drugs met with little success. Two world wars brought in the era of modern IV therapy. However, the greatest advance in drugs, equipment, and procedures has occurred in the past 25 years. PMID- 8708845 TI - Family caregivers' perspectives on hospice nursing care. AB - In hospice nursing, dying persons and their families are the unit of care and the focus of nursing interventions which are provided within the context of interdisciplinary care. The key component of hospice nursing is the interaction between the nurse, the terminally ill person, and the family. This naturalistic study was designed to describe the family caregiver's relationships between the hospice patient, the nurse, and themselves. The intent was to determine if family caregivers would develop empathic relationships with the hospice nurses. Nine bereaved family caregivers participated in the study. All were bereaved at least six months prior to being interviewed. The major finding of the study was that family caregivers perceived the hospice nurses as part of the family. The hospice nurses were characterized as respectful, kind, caring, clinical experts whose presence and interventions helped meet the needs of the family experiencing death. PMID- 8708846 TI - Assessing palliative care educational needs of physicians and nurses: results of a survey. Greater Victoria Hospital Society Palliative Care Committee. AB - The Greater Victoria Hospital Society (GVHS) Palliative Care Committee surveyed medical and nursing staff from four hospitals and The Victoria Hospice Society in February, 1993. The purpose of the survey was to identify physicians' and nurses' perceived educational needs related to death and dying. Programs that focus on the dying process; patient pain, symptom, and comfort control; and patient and family support were identified as necessary to meet the educational needs of physicians and nurses in providing quality palliative care. Physicians and nurses identified communication skills as being paramount. Communications concerning ethical issues were highlighted as the most difficult to cope with. PMID- 8708847 TI - Evaluating a palliative care program: methodology and limitations. AB - The article describes an evaluation of a palliative care service in a regional and tertiary care facility. The service components are described. The four outcomes chosen for evaluation were: (a) symptom relief; (b) satisfaction with care for patients/families; (c) utilization of community resources; (d) good nursing morale and low staff stress. Quality of life was measured using a symptom distress scale; satisfaction using an adapted Kristjanson FAMCARE scale; community resources with opinion and satisfaction surveys; and staff morale and stress with the Maslach Bumout Inventory and Latack's Coping Questionnaire. Results showed that overall symptom distress was reduced. Patients/families were generally satisfied, with some areas needing attention. Physicians were generally satisfied and believed patients/families benefited from the psychosocial support, respite, and education/information. Nurses felt they had the time, energy, resources and support to give quality care. PMID- 8708848 TI - When judges err. PMID- 8708849 TI - Palliative care providers' perspectives on service and education needs. AB - To obtain the information necessary for coordinated regional program development, we examined (a) the multidisciplinary viewpoint of palliative care service provision and (b) the continuing education needs reported by non-physician service providers. Of 146 surveys distributed to care providers from multiple settings, 135 were returned. Respondents cited these problems: fragmented services, poor pain and symptom control, lack of education for providers, lack of public awareness, problems with the continuity and coordination of care, lack of respite, and lack of hospice beds. Stress management for caregivers, pain management, communication skills, and symptom assessment were rated as priorities in continuing education. Lectures, small group discussions, practicum, and regular medical centre rounds were the preferred learning formats, while costs and staff shortages were cited as educational barriers. PMID- 8708850 TI - A questionnaire to medical students on palliative care and euthanasia. PMID- 8708852 TI - Successful management of intractable nausea with ondansetron: a case study. PMID- 8708851 TI - Assessment of the perception and worries [corrected] in terminally ill patients in Spain. PMID- 8708853 TI - Radiosurgical pituitary ablation for cancer pain. PMID- 8708854 TI - Prognosis in advanced pulmonary disease. PMID- 8708855 TI - Limits to multidisciplinary education. PMID- 8708856 TI - Dignity in dying: a preliminary study of patients in the last three days of life. AB - The final three days of life of 50 consecutive patients on a busy integrated palliative care service is described, with regard to final cause of death, symptom control, drug prescription, retention of personal function, and other measures possibly relevant to dignity in dying. Good symptom relief was maintained without rapid or high escalation of doses of morphine or sedatives. Personal function was maintained in at least a moderate degree in the majority of patients. This study also illustrates some of the difficulties in describing and evaluating the concept of "dying with dignity". PMID- 8708857 TI - Historical perspectives of cyclic vomiting. PMID- 8708858 TI - Cyclic vomiting syndrome. Introduction. PMID- 8708859 TI - The cyclic vomiting syndrome described. AB - Cyclic vomiting syndrome (CVS) consists of discrete episodes of nausea and vomiting lasting hours or days, separated by symptom-free intervals of similar or varying lengths. The disorder is not caused by underlying organic disease. The episodes tend to be similar in time of onset, duration, and symptomatology specific for each patient and are self-limited. The majority of patients can identify experiences or conditions that may precipitate episodes, the commonest being heightened emotional states and infections. The onset of CVS most often occurs during preschool or early school years, although it may begin at any age, from infancy to mature adulthood. CVS episodes can recur for months or decades, and their subsidence cannot be predicted. PMID- 8708860 TI - Differential diagnosis of cyclic vomiting syndrome. AB - Vomiting is a nonspecific symptom and cyclic vomiting is a symptom complex that may result from a disorder of any major organ system. Children with cyclic vomiting syndrome (CVS) need careful review and investigation at their earliest presentations to exclude potentially lethal abnormalities such as intestinal volvulus, metabolic disorders, and neurologic space-occupying lesions. The range of abnormalities that may present with features consistent with CVS includes gastrointestinal obstructive, inflammatory and motility abnormalities, pancreatic disease, metabolic disease (particularly the amino acid-opathies, organic acidurias, fatty acid oxidation defects, and acute intermittent porphyria), renal disease, epilepsy, migraine, and psychiatric disorders. Careful history taking will usually provide clues to these uncommon problems, but all children should undergo baseline assessment of gastrointestinal morphology and screening tests for renal and metabolic disease. PMID- 8708861 TI - Metabolic etiologies of cyclic or recurrent vomiting. AB - Treatable metabolic diseases are infrequently considered in the differential diagnosis of a child with cyclic or recurrent vomiting. Helpful clues in a complete medical history can heighten a clinician's suspicion about a metabolic etiology, as can the presence of certain symptom complexes or biochemical features. A rational approach to investigation is discussed using routine laboratory tests as a first-line screen. More definitive testing is indicated when screening results are abnormal or when the patient's history is particularly suggestive. Until test results become available, patients in whom a metabolic etiology cannot be ruled out should be treated as if they have an inborn error, in parallel with other conventional therapies, to avoid complications that are often preventable. PMID- 8708862 TI - The central control of vomiting. PMID- 8708863 TI - Serotonergic mediation of vomiting. AB - In the latter part of the 20th century, significant advances have been made in the understanding of the emetic reflex. As a result, there have been major improvements in the treatment of vomiting, particularly that associated with chemo- and radiotherapeutic treatments for neoplastic disease. The 5-HT3 receptor antagonists (ondansetron and granisetron) have been demonstrated to be of benefit in treating the profound emesis observed during cancer treatment. This observation, together with results from pharmacologic and physiologic investigations in both animals and humans, have identified 5-hydroxy-tryptamine (5-HT or serotonin) to be of fundamental importance in the pathogenesis of emesis. 5-HT appears to be released by radiation and chemotherapeutic agents from enterochromaffin cells within the wall of the intestine, and possibly from neurons within the brainstem. Stimulation of 5-HT3 receptors, located centrally in the dorsal medulla of the brainstem and peripherally on vagal afferent terminals in the gastrointestinal tract, appears to play a pivotal role in eliciting emesis. The interaction of 5-HT with non-5-HT3 receptors, particularly 5-HT1A and 5-HT4 receptors, may be important in the emetic reflex. The development of agents that interact with these receptors may offer alternative approaches to the treatment of nausea and vomiting. PMID- 8708864 TI - The role of the paraventricular system in the pathogenesis of cyclic vomiting. PMID- 8708865 TI - Psychophysiologic treatment of cyclic vomiting. AB - Cyclic vomiting (CV) presents a crisis not only to the identified patient but also to the entire family, who often feel like helpless victims of the effects of the vomiting cycles. This often creates despair, depression, and anger about the vomiting continuing indefinitely. If the child can be given effective medication and the family can be helped with the emotional burden surrounding the illness, the frequency and severity of CV tends to diminish. This article explores ways of lessening the psychologic and social effects of the cyclical vomiting through a multidisciplinary treatment plan for the family that involves the pediatrician, school, psychotherapist, and family as effective therapeutic agents. The child needs to be helped to express feelings which he or she cannot put into words. The article describes dramatic play involving symbolization of the illness, creation of metaphors in stories and drawings for the dilemmas surrounding CV and family therapy in which unnamed stresses are given consideration. Through this treatment, health and hope are promoted within each family member. PMID- 8708866 TI - Symptomatology and pathogenesis of migraine. AB - Migraine is a paroxysmal disorder of which headache is the central symptom. The headache is generally so intense that it is associated with other symptoms, such as nausea, vomiting, photo- and phonophobia. In so-called classic migraine or migraine with aura, the headache is preceded by transient focal neurological symptoms, generally referred to as aura symptoms. There is clinical experimental evidence for the involvement of at least three mechanisms in the pathogenesis of the migraine headache. These mechanisms are extracranial arterial vasodilation, extracranial neurogenic inflammation, and decreased inhibition of central pain transmission. The associated symptoms are possibly caused by sympathetic activation at a peripheral (nausea and vomiting) and central level (photo- and phonophobia). The aura symptoms are, in the author's opinion, most likely caused by a mechanism similar to spreading excitation and depression. An adaptation of the traditional concept of the pathogenesis of the migraine attack is presented. PMID- 8708867 TI - The relationship between cyclic vomiting syndrome and abdominal migraine. AB - Abdominal migraine and cyclic vomiting are both self-limiting episodic conditions of children, with periods of complete normality between episodes. The clinical features of both syndromes show considerable similarity, and resemble those found in association with migraine headaches. It is proposed that cyclic vomiting is a condition related to migraine. PMID- 8708868 TI - Neurologic investigations of childhood abdominal migraine: a combined electrophysiologic approach to diagnosis. AB - Neurologic studies of childhood migraine, including imaging techniques and electrophysiology, have identified a number of abnormalities, but none of high specificity. Therefore, a diagnostic marker for childhood migraine and its equivalents remains elusive. In this study a combined electrophysiologic approach is assessed, using measurements of visually provoked beta-activity, high frequency photic following responses, and visually stimulated event-related potentials. Eighteen children younger than 10 years of age with abdominal migraine and/or cyclic vomiting syndrome were investigated with these techniques. They were compared with the responses from a number of control groups, including a group diagnosed as childhood migraine with or without aura. Individually, each investigation yielded high sensitivities but low specificities ( < 50%). However, when these techniques were used in combination the specificity for migraine rose to > 90%. There was no significant difference between abdominal migraineurs and those with migraine with or without aura in any of the parameters studied. This combined approach is potentially useful in the diagnosis of abdominal migraine and supports the view that abdominal migraine can be classed as a true childhood migraine equivalent. PMID- 8708869 TI - Abdominal migraine as a cause of vomiting in children: a clinician's view. PMID- 8708870 TI - Management of cyclic vomiting syndrome. AB - The general goals of treatment of cyclic vomiting syndrome (CVS) are: interruption of established episodes, amelioration of symptoms in patients whose episodes cannot be interrupted, aborting episodes during prodromal symptoms, prophylaxis to abolish or lessen the frequency of episodes, and recovery. Complications of cyclic vomiting episodes include esophagitis, hematemesis, depletion of intracellular electrolytes, hypertension, and secretion of inappropriate antidiuretic hormone. PMID- 8708871 TI - Prophylactic therapy in cyclic vomiting syndrome. AB - The experience with prophylactic therapy for cyclic vomiting syndrome (CVS) at Princess Margaret Hospital for Children, Perth, Western Australia was retrospectively reviewed by questionnaire. Data was collected from 31 patients, aged 2.9-21.75 years who reported a mean of nine attacks per year. Eleven patients had utilized prophylactic therapy. Parental assessment of benefit was recorded. Propranolol was the most effective agent with reported benefit in four of six cases, other antimigraine agents were deemed effective in two of seven cases. Anti-convulsants and antidepressants were not considered useful. Homeopathic and vitamin supplements were thought to be beneficial in three of six cases. Prophylaxis was less likely to be beneficial in the more severe cases of CVS, but was of benefit in those patients whose attacks were precipitated by infection or who had features consistent with migraine. Prophylactic therapy with propranolol or serotonin receptor antagonists should be considered in children with frequent or severe symptoms. PMID- 8708872 TI - Cyclic vomiting: the pattern and syndrome paradigm. AB - Although it remains a mysterious disorder since its description over a century ago, cyclic vomiting syndrome appears to be more prevalent than previously thought. With the goal of improving detection of affected children, two quantitative historical criteria-peak intensity > or = 4 emeses/h) and a episode frequency < or = 9/ month-can differentiate those with an explosive, intermittent cyclic pattern from those with a low-grade, daily chronic pattern. Diagnoses that can cause a cyclic vomiting pattern includes abdominal migraine, chronic sinusitis, intracranial neoplasm, anomalies of and mucosal injury to the gastrointestinal tract, urologic abnormalities, and metabolic and endocrine disorders. Because many organic disorders can mimic cyclic vomiting syndrome, a systematic diagnostic evaluation of potential underlying disorders is recommended. The following terminology is proposed: a cyclic pattern is differentiated from a chronic pattern of vomiting on the basis of a higher peak intensity of emesis and lower frequency of episodes; and of those with a cyclic vomiting pattern, an idiopathic subgroup who is not found to have an underlying disorder on diagnostic testing can be labeled as having cyclic vomiting syndrome. PMID- 8708873 TI - Treatment of cyclic vomiting in childhood with erythromycin. AB - Cyclic vomiting syndrome (CVS) is an unusual cause of episodic emesis in children. It manifests as intermittent episodes of severe vomiting, similar in time of onset and duration, with no symptoms during the intervening period. Dehydration necessitating intravenous fluid therapy may occur. Most therapeutic maneuvers have proven unsuccessful. We report the use of erythromycin as a prokinetic agent in the treatment of cyclic vomiting in 24 children (10 boys, 14 girls). Many patients had mild associated abdominal pain with their vomiting. Fourteen patients had previously been given metoclopramide but none responded. Two patients were mildly developmentally delayed. Twenty-four patients were given oral erythromycin ethylsuccinate, approximately 20 mg/kg/day, in two to four divided doses for 7 days. This dose was repeated as needed when symptoms reappeared. Eighteen of 24 patients reported total resolution of symptoms when re evaluated at 2 and 6 months. All males responded, eight of 18 responders were female, and all six with partial or no response to therapy were female. This uncontrolled trial suggests that erythromycin may be a useful prokinetic agent in the treatment of CVS in childhood. Because the study was uncontrolled, placebo effect cannot be excluded. Case-controlled, double-blinded prospective trials should be considered to evaluate the effectiveness of erythromycin in CVS. PMID- 8708874 TI - The Shwachman Award of the North American Society for Pediatric Gastroenterology and Nutrition. Presentation: Shwachman Award. PMID- 8708875 TI - The Shwachman Award of the North American Society for Pediatric Gastroenterology and Nutrition. The Shwachman Award acceptance. PMID- 8708876 TI - Enhancing nutrition in cystic fibrosis with comprehensive therapies. PMID- 8708877 TI - Behavioral intervention to improve calorie intake of children with cystic fibrosis: treatment versus wait list control. AB - Changes in calorie intake and weight gain were evaluated in five children with cystic fibrosis (CF) who received behavioral intervention and four children with CF who served as wait list controls. The behavioral intervention was a 6-week group treatment that provided nutritional education plus management strategies aimed at mealtime behaviors that parents find most problematic. The control group was identified prospectively and was evaluated on all dependent measures at the same points in time pre- and posttreatment as the intervention group. Difference scores on calorie intake and weight gain from pre- to posttreatment were compared between groups using t tests for independent samples. The behavioral intervention group increased their calorie intake by 1,032 calories per day, while the control group's intake increased only 244 calories per day from pre- to posttreatment [t(6) = 2.826, p = 0.03]. The intervention group also gained significantly more weight (1.7 kg) than the control group (0 kg) over the 6 weeks of treatment [t(7) = 2.588, p = 0.03] and demonstrated catchup growth for weight, as indicated by improved weight Z scores (-1.18 to -0.738). The control group showed a decline in weight Z scores over this same time period (-1.715 to -1.76). One month posttreatment, the intervention was replicated with two of the four children from the control group. Improved calorie intake and weight gain pre- to posttreatment were again found in these children. At 3- and 6-month follow-up study of children receiving intervention, maintenance of calorie intake and weight gain was confirmed. No changes were found on pulmonary functioning, resting energy expenditure, or activity level pre- to posttreatment. This form of early intervention appears to be promising in improving nutritional status and needs to be investigated over a longer period of time to evaluate the effects of treatment gains on the disease process. PMID- 8708878 TI - Dietary intakes of young children with cystic fibrosis: is there a difference? AB - Caloric intakes of preadolescent and adolescent girls and boys with cystic fibrosis (CF) were compared in order to evaluate the possibility that poor caloric intake contributes to poor nutritional status and high mortality among girls with CF. Fifty-six CF patients (26 girls and 30 boys), 10-15 years old, completed a 3-day food record, answered a short questionnaire, and underwent anthropometric and pulmonary function assessment. The mean ages of the girls and boys were similar, but the height and weight percentiles of the girls were lower than those of the boys (p = 0.02). Mean caloric intakes were no different (116% and 112% of the recommended nutrient intake in the girls and boys, respectively). Nutritional status, as determined by weight as a percentage of ideal weight for height, mean triceps skinfold thickness, and midarm muscle circumference, was normal and similar in both sexes. Most girls and boys with CF in this study had an appropriate perception of their body weight. Pulmonary function tests suggested mild lung disease with no significant difference between girls and boys (forced expiratory volume in 1 s of 82.2% and 79.8% of predicted values, respectively). The similar nutritional and pulmonary status of the girls and boys with CF in this age group is in contrast to previous reports. This finding may be the result of our policy, introduced > 15 years ago, of expecting normal growth, by paying close attention to enzyme therapy and encouraging high energy intake from the time of diagnosis. It remains to be seen whether boys and girls continue to maintain similar nutritional and pulmonary status at a later age and whether both sexes experience a similar mortality rate as they age. PMID- 8708879 TI - Cisapride therapy for gastrointestinal disease. PMID- 8708881 TI - Effects of prednisolone and dexamethasone on alkali burns of the esophagus in rabbit. AB - Steroids have been widely used to prevent the severe secondary strictures that inevitably follow accidental ingestion of caustic substances by children, though with controversial results. To confirm the efficacy of large early doses of dexamethasone, we conducted an experimental study in rabbits. A cotton swab soaked in sodium hydroxide was placed on the esophageal mucosa of 30 rabbits. The rabbits were then treated with antibiotics and randomly allocated to one of three groups. One group received no steroids; the second group was treated with prednisolone; and the third was given dexamethasone. After 21 days, the rabbits were killed and the severity of the lesion was evaluated by radiological, anatomical, and histological examination. Rabbits with moderate or severe stricture, as defined on the basis of radiological examination, made up 80% of the control group, 70% of the prednisolone-treated group, and 40% of the dexamethasone-treated group. Rabbits with third-degree burns, with or without macroscopic ulcers, made up 60% of the control group, 40% of the prednisolone treated group, and 30% of the dexamethasone-treated group. On histological examination, rabbits with third-degree lesions made up 70% of the control group, 40% of the prednisolone-treated group, and only 20% of the dexamethasone-treated group. Only the difference between the control and dexamethasone-treated group was statistically significant. Taken together, our results clearly indicate that, in rabbits, rapid postburn administration of dexamethasone leads to significant reductions in the frequency and severity of strictures and in the severity of burns. PMID- 8708880 TI - Long-term persistence of hepatitis B virus DNA in the serum of children with chronic hepatitis B after hepatitis B e antigen to antibody seroconversion. AB - To evaluate the prevalence and duration of viremia in relation to the features of liver disease, we investigated hepatitis B virus (HBV) DNA by the polymerase chain reaction in the serum of 39 children with chronic hepatitis B, after hepatitis B e antigen to antibody seroconversion. During a mean observation period of 8.2 +/- 3.8 years after seroconversion, all patients were asymptomatic; 36 had persistently normal alanine aminotransferase levels, and three had occasional mild alterations. Liver histology, checked in 21 patients, showed persistent hepatitis in nine, fibrosis in 10, and cirrhosis in two cases. HBV DNA was always undetectable by dot blot hybridization. Five children eventually cleared hepatitis B surface antigen, including one with cirrhosis who developed liver cancer at 19 years. HBV DNA was detected by polymerase chain reaction in 87% of children within 5 years of follow-up, in 58% of cases 6-10 years after seroconversion (p < 0.001), and in 50% of patients investigated later. Long-term viremia was found in two patients (40%) who cleared HBsAg, including the one who developed liver cancer. The chances of clearing viremia during follow-up were higher in children with acute hepatitis at the onset of illness (86%) than in those with asymptomatic onset (37%; p < 0.05). Our results show that low levels of HBV viremia, probably reflecting low levels of virus replication, persist for several years in children with chronic hepatitis B after hepatitis B e antigen to antibody seroconversion and remission of liver disease, even after the clearance of hepatitis B surface antigen. Persistent replication could support mild biochemical alterations and inflammatory liver lesions. It could allow late reactivation of liver disease and may play a role in the development of carcinoma. PMID- 8708882 TI - Sucrase-isomaltase deficiency: changing pattern over two decades. AB - Twenty-two cases of sucrase-isomaltase deficiency (SID) were observed over a period of 20 years. Since 1977 delay of introduction of sucrose and its decrease in infants' diets have modified the symptomatology. In general, onset of diarrhea has not taken place immediately but 15 days to 2 months after introduction of sucrose. Out of 12 cases with dehydration, five occurred 3 to 7 months after the beginning of sucrose diet. Hypotrophy was not constant (11 of 22 cases), thus diagnosis was delayed in 17 of 22 cases. A yellow complexion due to rising carotene levels in the blood is a striking feature. Because of falsely positive sucrose load tests (four out of 14 nonSID infants) and failure of the hydrogene breath test (one out of five studied cases), disaccharidase determination remains the key to diagnosis. Despite the genetic difference symptoms seem to depend on infant feeding practices. PMID- 8708883 TI - Thiamine, riboflavin, folate, and vitamin B12 status of infants with low birth Weights receiving enteral nutrition. AB - The purpose of the present study was to monitor the vitamin status of 14 low birth-weight (LBW) infants (< 1,750 g birth weight) at 2 weeks and an additional four infants at 3 weeks who were receiving an enteral formula providing 247 micrograms/100 kcal thiamine, 617 micrograms/100 kcal riboflavin, 37 micrograms/100 kcal folate, and 0.55 micrograms/100 kcal vitamin B12. The mean birth weight of the 18 infants was 1,100 +/- 259 g, and mean gestational age was 29 +/- 2 weeks. Weekly blood, 24-h urine collections, and dietary intake data were obtained. For thiamine, red blood cell (RBC) transketolase activity was within the normal range for all infants. For riboflavin, RBC glutathione reductase activity was normal for all infants except one. We calculated from intake and urinary excretion data that these infants require 225 micrograms/100 kcal thiamine and 370 micrograms/100 kcal riboflavin, respectively. Mean plasma folate levels were 21 +/- 11 ng/ml at 2 weeks and 18 +/- 5 ng/ml at 3 weeks. RBC folate levels were 455 +/- 280 ng/ml at 2 weeks and 391 +/- 168 ng/ml at 3 weeks. All folate blood values were normal, except for one subject with an elevated level (59 ng/ml). Vitamin B12 plasma values were 737 +/- 394 pg/ml at 2 weeks and 768 +/- 350 pg/ml at 3 weeks, and all values were normal except for three infants with elevated values. In conclusion, appropriate vitamin status was maintained during this short observational period, during administration of this enteral formula; however, riboflavin concentrations in the enteral feed may be excessive. PMID- 8708884 TI - Combined use of cyclosporine and azathioprine or 6-mercaptopurine in pediatric inflammatory bowel disease. AB - The aim of this study was to assess whether in steroid-resistant patients with pediatric inflammatory bowel disease (IBD) a combination of cyclosporine and azathioprine (or 6-mercaptopurine) could induce remission and subsequently permit maintenance on azathioprine/6-mercaptopurine as the sole immunosuppressive agent. Two boys and six girls (six with ulcerative colitis and two with Crohn's disease; ages 3-17 years) received 100-200 micrograms/kg/day cyclosporine intravenously and then 4-10 mg/kg/day orally. Doses were adjusted to achieve trough serum cyclosporine levels of 100-200 mu/L (Abbott's TDX assay). Seven of the eight patients received azathioprine/6-mercaptopurine, and all were given a 5 aminosalicylate preparation and corticosteroids. The latter drugs were continued and then tapered off as clinical status allowed. Cyclosporine was continued for 3 10 months in those who responded. In seven of eight patients, there was a rapid clinical response; one patient showed a transient response, but recurrent bleeding led to total colectomy 9 days after starting cyclosporine. Of the seven responders, three were able to discontinue prednisone and cyclosporine and are doing well on azathioprine at long-term follow-up (2-5 years). One who did not receive azathioprine/6-mercaptopurine maintained remission for 2 years after cyclosporine was stopped, one experienced a disease flare-up 5 months after start of cyclosporine treatment and required colectomy, one who did not tolerate 6 mercaptopurine had a flare-up during cyclosporine tapering and underwent surgery at 6 months, and one started to flare up with cyclosporine tapering at 6 months and was scheduled for surgery. No significant complications of treatment were observed. Seven patients had an initial response and four of them have so far not required surgery. These preliminary findings suggest that azathioprine/6 mercaptopurine can be used safely to maintain cyclosporine-induced remission in children with IBD. PMID- 8708885 TI - Randomised trial of effect of delayed intravenous lipid administration on chronic lung disease in preterm neonates. AB - A recent sevenfold increase in the annual incidence of chronic neonatal lung disease (CNLD) on an intensive care unit was attributed to the early administration of intravenous lipid (IVL) in ventilated preterm neonates. When logistic regression was used to eliminate other confounding variables, early delivery of IVL was independently associated with an eight-fold increase in the likelihood of CNLD. Consequently, we designed a prospective study to detect a halving of the incidence of CNLD by delaying IVL administration from 5 days (as is routine practice) to 14 days. Sixty-four parenterally fed preterm neonates weighing < 1,500 g at birth were randomised to receive IVL either on day 5 or day 14. Analysis was by intention to treat, since several infants in the latter group required no parenteral nutrition by day 14. Our results showed that the relative risk (95% confidence interval) of CNLD at 28 postnatal days was 1.15 (0.81-1.62); at 36 weeks postconception, it was 1.08 (0.59-1.99). A study population of > 2,000 would be required to determine whether these relative risks were significantly different from 1. PMID- 8708886 TI - Increase in density of jejunal IgA2 cells during infancy without change in IgA1 cells. AB - To determine any association between densities of cells containing IgA, IgA1, IgA2, IgG, and IgM in the lamina propria of the jejunum and the age of the individual, we restudied 65 specimens from 65 patients in the age range 66 days to 4 years. Indications for jejunal biopsy were minor symptoms; none of the patients had any gastrointestinal disease, and the specimens showed normal morphology. We used murine anti-human monoclonal antibodies to IgA, IgA1, IgA2, IgG, and IgM and an indirect immunoperoxidase method and found a significant increase with advancing age in the densities of cells containing IgA, IgA2, and IgG, whereas the densities of IgA1- and IgM-positive cells did not change with age. Densities of IgA2 and IgG cells in children over 1 year were significantly greater than in infants under 9 months (upper vs. lower quartile of the study group, p < 0.05 in both comparisons). In the jejunum, an increase in IgA2 cells was responsible for the increase in total IgA production for the age range studied. PMID- 8708887 TI - Development of iron status and response to iron medication in pubertal boys. AB - During male puberty, rapid growth of body mass and the rise in hemoglobin concentration require much iron. The increasing need for iron may carry a risk of iron deficiency. Criteria for iron deficiency in adolescence are difficult to assess, as indicated by prevalences ranging from 0.5 to 30% in five recent studies from developed countries. We followed 60 prepubertal or early pubertal boys, testing them at 6-month intervals for 24 months and relating hemoglobin levels and other iron parameters to age, pubertal development, and response to iron medication. A significant increase in mean hemoglobin was first seen at genital stage G4. In early puberty, in contrast, between stages G1 and G3, ferritin decreased. Mean transferrin increased slightly during the study. The prevalence of iron deficiency increased both with age and with advancing puberty. However, the early decline in ferritin may be a physiologic response to guarantee increasing intestinal absorption. After iron medication, we observed improvements in the levels of our parameters of iron status, which showed that hemoglobin levels had been limited by iron deficiency. In pubertal boys, evaluation of iron deficiency should not be based on laboratory parameters alone; age, pubertal development, and growth should also be taken into account. PMID- 8708889 TI - Macroscopic versus microscopic diagnosis of reflux esophagitis: erosions or eosinophils? PMID- 8708890 TI - Reflux esophagitis: biopsy or not? PMID- 8708888 TI - Recurrent vomiting following Harrington rod instrumentation of the spine. PMID- 8708891 TI - Allergy to soy lecithin in a child. PMID- 8708892 TI - Candida esophagitis in a child with achalasia. PMID- 8708893 TI - Tension pneumoperitoneum: a potentially life-threatening complication of percutaneous endoscopic gastrojejunostomy. PMID- 8708894 TI - Torticollis as a side effect of cisapride treatment in an infant. PMID- 8708895 TI - IGF-1. PMID- 8708896 TI - Helicobacter pylori-specific IgM antibodies in children born to infected mothers. PMID- 8708897 TI - Etiology and outcome of pediatric burns. AB - A 6-year retrospective review of burn victims hospitalized at a major burn center was conducted to determine the etiology and outcome of pediatric burns. Four hundred forty-nine patients under age 16 years were identified and stratified by age, sex, burn size, presence or absence of inhalation injury, cause of burn, and county of residence. The mean patient age was 4.3 +/- 0.2 years, and the male:female ratio was 1.9:1. There were 21 deaths overall (4.7%), the majority of which (18) were among children under 4 years of age. With respect to large burns, defined as > and = 30% total body surface area (TBSA), the mortality rate for children under age 4 was significantly higher than that for older children (46.9% v 12.5%; P < .01), despite the nearly identical mean burn size of the two groups. Except for burn incidence, there were no significant differences between males and females. The mean burn size was 15.1% +/- 0.7%, and was significantly larger for nonsurvivors than survivors (55.3% +/- 5.7 v 13.1% +/- 0.5%; P < .01). Inhalation injuries were strongly associated with large burns and were present in all 15 flame-burn fatalities. Scalds were the most common type of burn among children under 4 years of age; flame burns predominated in older children. There were 6 deaths related to scalds, all of which occurred in children under 4. Burn type, size, and mortality rate did not differ between children from urban and rural counties. Large burn size was the strongest predictor of mortality, followed by (in order) age less than 4 and the presence of inhalation injury. Infants and young children have the highest risk of death from burn injury. Burns smaller than 30% TBSA without an inhalation injury (such as small scald injuries) occasionally are lethal in infants and small children, despite modern therapy. PMID- 8708898 TI - Balloon dilation of esophageal strictures in children. AB - Esophageal strictures in children may develop as a primary constriction, secondary to a surgically repaired esophageal atresia (with or without tracheoesophageal fistula), as a result of chemical injury after caustic ingestion, or following esophageal surgery. Traditional treatment of esophageal strictures has been limited to dilation (using bougie dilators) with esophagoscopy under general anesthesia. Recent reports have shown success with fluoroscopically guided balloon catheter dilation. Eight children (aged 2 months to 14 years) were treated with balloon catheter dilation for focal strictures of the esophagus. In six of the eight cases, complete resolution of the strictures was achieved after an average of 7.5 dilations (range, 1 to 14). Two of the eight patients moved to another part of the country and did not complete treatment. There has been no morbidity or mortality. In selected centers, balloon catheter dilation under fluoroscopic guidance has become a safe treatment of benign esophageal strictures in children. It should be considered the treatment of choice in the initial management of esophageal narrowing and appears to be safer than the more traditional methods of esophageal dilation. PMID- 8708899 TI - Intratracheal pulmonary ventilation provides effective ventilation in a near drowning model. AB - Overdistension of the lungs from high inspiratory pressure is increasingly recognized as a major contributor to lung injury and worsening respiratory failure in the child who requires prolonged mechanical ventilation. Many modes of ventilation (such as high-frequency ventilation) have been introduced in an attempt to decrease this lung injury. Recently, a new mode of tracheal ventilation, intratracheal pulmonary ventilation (ITPV), has been described. By using a catheter positioned at the carina with continuous gas flow, it is possible to achieve effective ventilation at very low pressures. The purpose of this study was to evaluate the usefulness of ITPV in a near-drowning model. Ten domestic Yorkshire swine underwent arterial, venous, and pulmonary arterial catheter as well as tracheotomy placement. All animals received 13 mL/kg of fresh water intratracheally to induce a pulmonary injury. Six pigs were ventilated for 4 hours using ITPV; the other four pigs received conventional mechanical ventilation (CMV). Circulatory and ventilatory pressures, hemodynamic variables, arterial blood gases, and end-tidal CO2 were measured before lung injury and every 30 minutes thereafter. Both proximal and distal peak and mean airway pressures were measured. The animals were ventilated as needed to maintain the arterial blood gases in the normal range. The authors found the expected changes in pulmonary compliance, oxygen requirement, and airway pressure after inducement of lung injury. The six animals treated with ITPV had significantly lower airway pressures than those of controls. Peak inspiratory pressures with ITPV were 8.2 +/- 1.9 cm H2O versus 17.8 +/- 3.7 with CMV (P < .001). Distal mean airway pressures using ITPV were 2.3 +/- 0.1 cm H2O versus 9.0 +/- 3.2 with CMV (P < .01). With respect to hemodynamic variables, there were no differences between experimental and control animals. In conclusion, ITPV can afford effective ventilation in a near-drowning model of lung injury at airway pressures significantly lower than those required with CMV. ITPV could be a very valuable addition to the currently available methods of mechanical ventilation. PMID- 8708900 TI - Gastroschisis bowel in an intact exomphalos: implications for etiology and possible prevention. AB - The etiology of changes in the bowel in gastroschisis is unknown. A case of exomphalos is described, in which the bowel had a gastroschisis-like appearance. The possibility of a vascular mechanism as the cause is discussed. PMID- 8708901 TI - Coloplasty for congenital short colon. AB - Seventy-two patients with complete short colon (CSC) were treated at the authors' institution between 1971 and 1994. Initially, one-stage coloplasty with abdominoperineal pull-through was performed in 10 neonates, with a 50% survival rate. The subsequent 62 patients had staged management in which a window colostomy was performed at the time of presentation. Of the 50 survivors in this group, 20 had coloplasty with a abdominoperineal or sacroabdominoperineal pull through by June 1994. A protective ileostomy was done at the time of coloplasty in 10 patients, and was closed 6 to 12 weeks later. Two of the 20 patients who had staged reconstruction died because of complications; the others recovered well. The overall survival rate after coloplasty was 76.7%. Eighteen of the 23 survivors have had follow-up for up to 8 years. They have shown normal growth and good continence and have been free of significant long-term complications. The other 30 patients are awaiting definitive surgery. A staged management plan is advocated for complete short colon because it provides consistent results and good quality of life. PMID- 8708902 TI - Development in lung function of the affected side after repair of congenital diaphragmatic hernia. AB - The widespread use of newly developed techniques including extracorporeal membrane oxygenation (ECMO) has led to the survival of a number of patients with congenital diaphragmatic hernia (CDH) and associated hypoplastic lung. However, it is not fully recognized whether the hypoplastic and small lung of the affected side has the ability to develop its function after repair of CDH. The authors studied the lung function of 32 patients with CDH in whom these new methods were used. Two parameters, lung volume and pulmonary perfusion amount, were used to evaluate lung function. The former (checked by computed tomography scan) was used to evaluate the size of lung; the latter (checked by perfusion scintigram) was used to assess vascular density. The patients were divided into two groups, based on values of alveolar-arterial difference in oxygen content (AaDo2) at the time of admission. In group A (AaDo2 < 500 mm Hg; 12 cases), whose respiratory distress was mild and could be managed with ventilator care alone, the mean lung volume value for the affected side was 86% of the contralateral lung value from the initial study, and reached 93% at the time of follow-up study. The perfusion amount also exceeded 80% of the contralateral lung value from the initial study. Thus, it is likely that group A's affected-side lung is not small and has developed at a rate similar to that of the contralateral lung. However, in group B patients (AaDo2 > 500 mm Hg; 20 cases), who had severe respiratory distress at the admission and were managed with new techniques including ECMO, both lung volume and perfusion amount of the affected side initially were low in all cases (ie, mean values were 61% and 53% of contralateral-lung values, respectively). At the time of follow-up, the lung volume had increased in most cases (mean value, 88% of the contralateral lung value), but the perfusion amount of the affected side had not increased in most cases. It remained low, or decreased to below the initial value; the mean was 53% of the contralateral lung value. The initial mean perfusion: volume ratio (87%) had decreased significantly (to 62%) by the time of follow-up. This tendency was exaggerated in the 11 ECMO cases. These data might indicate that in most group B cases, the lung of the affected side has little ability to develop arterial branches, or certainly will be delayed in comparison to the contralateral lung, and that enlargement of lung volume may depend on overexpansion or emphysematous change rather than cellular growth. The present data also suggest that, in group B cases, total lung function will depend on the contralateral lung for a relatively long time. PMID- 8708903 TI - Innervation patterns of the rectal pouch and fistula in anorectal malformations: a preliminary report. AB - The innervation patterns of the rectal pouch and fistula of 52 children with anorectal malformations were investigated. Posterior sagittal anorectoplasty was used for intermediate and high anomalies; for the latter it was combined with an abdominal approach. Perineoproctoplasty was performed for low anomalies. The specimens were investigated by acetylcholinesterase staining, lactate dehydrogenase, and succinyldehydrogenase reaction. They consisted of fistula material only in 23 patients and of parts of the rectal pouch in 29. Fourty-four patients (84.6%) had follow-up, and information of bowel movements and continence was obtained after a mean of 3.3 years. Abnormal innervation patterns were found in 96% of the specimens. All fistulas were found to be aganglionic, including the adjacent part of the rectum involving the internal sphincter equivalent. Classical aganglionosis was found in 31% of the rectal pouch specimens, hypoganglionosis in 38%, neuronal intestinal dysplasia (NID) type B in 14%, and dysganglionosis in 10%. All patients with severe constipation or soiling at the time of follow-up had some histopathological correlation. Of the 25 patients for whom the specimens had consisted of rectal pouch material, nine (31%) had severe constipation. All four patients with a low-type malformation who had follow-up and pathological innervation patterns in the rectal pouch suffered from severe constipation; this was true of only five of the 19 children with intermediate or high malformations (P < .05). However, numerous pathological innervation patterns had been identified in patients who had normal bowel function at the time of follow-up. It is concluded that partial denervation of the rectum may not be the only cause in the pathogenesis of constipation after posterior sagittal anorectoplasty and perineoproctoplasty. The high frequency of neuronal intestinal malformations in the rectal pouch may be related to the higher frequency of bowel disturbances in patients with low malformations, in whom the resection was less radical. However, the clinical course is not necessarily related to specific histopathological findings. In the authors' opinion, the recommendation to use the distal rectal pouch and parts of the fistula in the reconstruction of anorectal malformations should be reconsidered. PMID- 8708904 TI - 'Triangular cord': a sonographic finding applicable in the diagnosis of biliary atresia. AB - Biliary atresia (BA) is characterized by luminal obstruction of the extrahepatic bile duct with fibrous remnants. The authors reviewed ultrasonographic examinations of the fibrous tissue in the bifurcation of the portal vein at the porta hepatis and identified the triangular- or tubular-shaped echogenic density, the so-called "triangular cord" (TC), in the vicinity of the portal vein on a transverse or longitudinal scan. In this prospective study, the authors investigated whether TC was useful in the noninvasive diagnosis of biliary atresia in 18 infants who had persistent neonatal jaundice. This was done by comparing the ultrasonographic examination with the histopathologic examination (HPE) of liver specimens obtained from a needle biopsy. The TC was identified in nine patients, all of whom were confirmed to have BA by HPE. The TC was not observed in the other nine patients, who had neonatal hepatitis (NH). The mean size of the TC was 13 mm (wide) x 6 mm (thick) (width range, 5 to 21 mm; thickness range, 4 to 12 mm). The diagnosis of BA was confirmed at the time of Kasai hepatoportojejunostomy in eight of the nine patients whose TC was noted by ultrasonography (US). The other patient was discharged because his parents refused the operation; he died of liver failure at 15 months of age. The nine patients with absent TC were treated medically for NH. Eight of them improved clinically. The other, diagnosed to have NH by needle and wedge liver biopsies, was reexamined 40 days after the initial examination because of worsening jaundice. A 18 x 12-mm TC was visualized ultrasonographically. Additionally, a percutaneous liver biopsy specimen showed BA with severe portal fibrosis and ductal proliferation. The patient underwent a Kasai hepatoportoenterostomy. On the basis of these results, the authors conclude that TC is a very specific ultrasonographic finding, representing the fibrous cone at the porta hepatis, and is a useful tool in the noninvasive diagnosis of BA. However, early exploration or close US follow-up is recommended for any patient suspected of having BA clinically, even if a liver biopsy confirms the NH. PMID- 8708905 TI - Electron microscopic study of the liver with biliary atresia and neonatal hepatitis. AB - Eleven cases of biliary atresia (BA) and eight of neonatal hepatitis (NH) were studied, using transmission electron microscopy, to document their different ultrastructural characteristics and to elucidate the possible pathogenesis of biliary atresia. Among 30 consecutive liver biopsies obtained from 19 infants with BA or NH, 21 specimens composed (13 BA, 8 NH) were examined ultrastructurally. The electron microscopic features of NH (patients' age range, 35 to 60 days) were (1) giant hepatocytic transformation with scattered areas of dilated endoplasmic reticulum, indicative of intracytoplasmic degeneration, (2) frequent cytoplasmic biliary necrosis, and (3) relatively intact microvilli in most bile canaliculi, which contained some hepatocytic cytoplasmic fragments. These features strongly suggest that the main pathological process in NH is hepatocellular injury rather than bile duct damage. In contrast, all cases with BA (age range, 27 to 130 days) demonstrated (1) marked hepatocellular cholestasis associated with many lysosomes and myelin figures, (2) marked loss of bile canalicular microvilli, (3) degenerated bile ductular cells containing bile pigments, and (4) periductal inflammatory fibrosis. These features suggest that the main pathological process in BA involves the biliary system. A few viral inclusions were observed in two cases with BA, which suggests that viral infection is a potential cause. In two BA cases (aged 40 and 43 days at the time of first biopsy), the ultrastructural findings essentially were the same as those of NH, and follow-up biopsy specimens (at 48 and 94 days) showed findings consistent with BA. Such results support Landing's hypothesis that BA and NH are different manifestations of a single pathological process. PMID- 8708906 TI - Care-giver evaluation of anti-gastroesophageal reflux procedures in neurologically impaired children: what is the real-life outcome? AB - Although the pediatric surgical literature is replete with reports of the success of operations for gastroesophageal reflux, postoperative complications are being reported with increasing frequency for the neurologically impaired subpopulation. Because a large portion of a care-giver's life is involved in attending to a neurologically impaired child, parental satisfaction with the outcome of these operations should be an important consideration when the use of such procedures is contemplated. The purpose of the present study was to assess the impact of antireflux operations with respect to care-giver opinions regarding the procedure. The authors retrospectively reviewed 25 charts (of 13 girls and 12 boys; age range, 3 months to 18 years) and documented (through survey results) perceived child well-being, objective care requirements, and overall care-giver satisfaction with the procedure. Results indicate there was significant improvement in feeding indexes, care-giver perception of the child's comfort, and quality of life postoperatively. Moreover, there was significant improvement in the care-givers' attitudes regarding their child, including the level of frustration in caring for the child, and the parents' overall quality of life. Care-givers also believed that the operation's result was about or better than what they had expected. In conclusion, the study documents care-giver satisfaction with antireflux procedures. Postoperatively, child care is easier and the quality of time spent with the child is better. The impression of better quality of life postoperatively for a neurologically impaired child may be the greatest success in this sometimes frustrating endeavor. PMID- 8708907 TI - Current status of 21 patients who have survived more than 20 years since undergoing surgery for biliary atresia. AB - Between 1952 and 1993, 289 patients with biliary atresia underwent surgery at the authors' institution. Twenty-two of them survived more than 20 years; one has since died of hepatic failure (at age 28 years). Of the 21 current survivors (age range, 20 to 39 years), 13 underwent hepatic portoenterostomy; the others had hepaticoenterostomy. None of these patients has undergone liver transplantation. Sixteen patients have led near-normal lives. This includes three married women, one of whom has given birth to a healthy baby boy. Of the six patients who had portal hypertension, three underwent both splenectomy and proximal splenorenal shunting in or before 1985. None of these patients has required additional treatment for portal hypertension. The quality of life of one patient has been severely affected by an unrelated condition (Turner's syndrome). A 22-year-old man was diagnosed as having intrahepatic stones 3 years ago. In another 22-year old man, hepatic dysfunction developed after frequent episodes of cholangitis. He is now being considered for liver transplantation. The majority of the long-term survivors have good quality of life. However, a few continue to suffer from complications including recurrent cholangitis. Close long-term postoperative follow-up is required for patients with biliary atresia. PMID- 8708908 TI - Laparoscopic surgery for neuroblastoma identified by mass screening. AB - Most neuroblastoma identified by mass screening are clearly different from the clinically diagnosed tumors with respect to biology, epidemiology, and outcome. Because the neuroblastomas detected by mass screening have favorable biological features (97% survival rate at 5 years), aggressive surgery may not be appropriate for such tumors. Laparoscopic adrenalectomy was performed on a 9 month-old boy and two 8-month-old girls who had suitable lesions. The tumors were small (< 20 mm in diameter). With the patients under general anesthesia, five trocars were placed in the abdomen. The abdominal wall traction method was used in addition to pneumoperitoneum. Intraabdominal pressure was maintained at below 4 mm Hg. The adrenal tumors were well encapsulated and completely excised, placed into a plastic bag, and removed through one of the 10-mm trocar sites. No lymphadenopathy was observed. The postoperative course was uneventful. The tumors were of favorable Shimada histology and had no N-myc gene amplification. The patients have survived, with no evidence of recurrence or metastasis, through the follow-up period of 17 to 22 months. PMID- 8708909 TI - Sustained pulmonary vasodilation after inhaled nitric oxide for hypoxic pulmonary hypertension in swine. AB - It has been shown that pulmonary vasodilation is sustained after discontinuation of inhaled nitric oxide (INO) during moderate hypoxic pulmonary hypertension (HPH) in swine. The present investigations demonstrated how INO dose, hypoxia duration, and endogenous NO production influence this important phenomenon. Fifteen adolescent Yorkshire swine were randomly assigned to three groups (n = 5 each) and underwent the following phasic experimental protocol: (I) Baseline ventilation (FIO2 = .3); (II) Initiating HPH (FIO2 = .16 to .18, PaO2 = 45 to 55 mm Hg); (III) INO at 10 ppm; (IV) Posttreatment observation; (V) INO of 80 ppm; and (VI) Posttreatment observation. Phase II (pretreatment hypoxia) lasted 30 minutes in group A (short hypoxia) and 120 minutes in group B (long hypoxia). N nitro-L-arginine methyl ester (NAME) was used to inhibit nitric oxide synthase (NOS) throughout the experiment in group C (short hypoxia + NAME). Hemodynamics and blood gases were monitored by systemic and pulmonary artery catheters placed by femoral cutdown. Analysis of variance with post-hoc adjustment was used to compare groups at each phase, and the paired t test was used for comparisons within a group. With respect to baseline mean pulmonary artery pressure (MPAP) and pulmonary vascular resistance (PVR), there were no significant differences among the three groups. MPAP and PVR were significantly higher in group C than in group A during phase II, (MPAP, 76% +/- 8% v 33% +/- 2%; PVR, 197% +/- 19% v 78% +/- 10%; P < .05). There were no significant differences in MPAP or PVR during phases III through VI. When MPAP was expressed as percent dilation, 80 ppm caused significantly more dilation than did 10 ppm in all three groups. Groups A and C had significantly higher sustained pulmonary artery dilation after 80 ppm than after 10 ppm (A, 82% +/- 31% v 17% +/- 11%; C, 68% +/- 10% v 42% +/- 12%; both P < .05), but group B did not (43% +/- 15% v 30% +/- 9%; P = .25). High dose results in stronger vasodilation than low dose during and after INO for moderate HPH of short duration. Long hypoxia blunts this high-dose advantage. Endogenous NO inhibition augments HPH but does not decrease pulmonary vasodilation during or after INO. PMID- 8708910 TI - Pyomyositis in children, caused by anaerobic bacteria. AB - The author describes the microbiology and clinical features of six pyomyositis infections in children, which yielded anaerobic bacteria. Anaerobic bacteria alone were recovered in four instances, and they are mixed with facultative bacteria in two. There were 15 bacterial isolates (13 anaerobic, 2 facultative). The bacteria were Peptostreptococcus sp (5 isolates), Bacteroides fragilis (3), Clostridium sp (2), Fuso-bacterium nucleatum (1), Prevotella sp (1), Bateroides sp (1), Streptococcus pyogenes (1), and Escherichia coli (1). Recent trauma or injury had occurred in five cases; three such injuries were from penetrating objects. This study highlights the potential importance of anaerobic bacteria in children with pyomyositis. PMID- 8708911 TI - In utero repair of experimental myelomeningocele saves neurological function at birth. AB - In a previous series of fetal sheep experiments, the authors demonstrated that midgestational exposure of the normal spinal cord to the amniotic space leads to a myelomeningocele (MMC) at birth that closely resembles human MMC phenotypes in terms of morphology and functional deficit. The present study tested whether delayed in utero repair of such evolving experimental MMC lesions spares neurological function. In 12 sheep fetuses, a spina bifida-type lesion with exposure of the lumbar spinal cord was created at 75 days' gestation (full term, 150 days). Four weeks later, the developing MMC lesions were repaired in utero for seven fetuses (five fetuses died before this time). Of those that had repair, three were delivered near term by cesarean section, and four died in utero or were aborted. All survivors had healed skin wounds and near-normal neurological function. Despite mild paraparesis, they were able to stand, walk, and perform demanding motor tests. Sensory function of the hindlimbs was present clinically and confirmed electrophysiologically. No signs of incontinence were detected. Histologically, the exposed and then covered spinal cord showed significant deformation, but the anatomic hallmarks as well as the cytoarchitecture of the spinal cord essentially were preserved. These findings show that timely in utero repair of developing experimental MMC stops the otherwise ongoing process of spinal cord destruction and "rescues" neurological function by the time of birth. Because there is evidence that a similar secondary damage to the exposed neural tissue also occurs in human MMC, we propose that in utero repair of selected human fetuses might reduce the neurological disaster commonly encountered after birth. PMID- 8708912 TI - The intussusception antireflux valve is ineffective in preventing cholangitis in biliary atresia. AB - Cholangitis is the most common complication after portoenterostomy for biliary atresia. The construction of an intussusception valve in the Roux-en-Y limb of the portoenterostomy has been advocated as a means to ameliorate this complication. The authors reviewed the records of children who underwent portoenterostomy to assess the incidence and severity of cholangitis, as well as outcome. The children were divided into two groups according to whether they did not have (group I) or did have (group II) an antireflux valve in the Roux limb of the bilioenteric conduit. There were 19 children in each group. There were no significant differences between the groups with respect to age at time of portoenterostomy or length of follow-up (Table 1). Nine group I patients are alive and well; five died and five have had or are awaiting transplantation. In group II, 10 patients are alive and well, eight have had transplantation, and one is awaiting transplantation. Outcome with respect to death or the need for liver transplantation because of progressive hepatic failure is not different between the groups (P = 1.0, Fisher's 2 x 2). The incidence of cholangitis was evaluated by comparing the average number of episodes of cholangitis during the follow-up period, the number of episodes per year, and the number of episodes in the first postoperative year (when this complication is most prevalent). Analysis of the data showed no difference in incidence between those with valved and nonvalved biliary conduits. The severity of cholangitis, judged by total length of antibiotic treatment, did not differ between the groups. It appears that the presence of an intussusception valve in the Roux-en-Y biliary conduit does not affect the incidence of cholangitis or the outcome after portoenterostomy, over short-term follow-up. PMID- 8708913 TI - Urologic problems in anorectal malformations. Part 1: Urodynamic findings and significance of sacral anomalies. AB - A series of 90 children with anorectal malformations received a total of 163 urodynamic studies. The mean number of studies per patient was 1.8 (range, 1 to 7), and the mean age at the time of initial evaluation was 17 months (range, 1 to 128). Sacral anomalies were found in 38 children (42%); 30 of them had partial sacral agenesis and eight had sacral dysplasia without bony defect. Normal lower urinary tract function was found in 98% of the children who had a normal sacrum, in all children who had sacral dysplasia, and in a small number of those who had partial sacral agenesis that affected smaller parts of the S-4 and S-5 pedicles. Severe lower urinary tract dysfunction was observed in 22 patients (24%), all but one of whom had partial sacral agenesis. There was no correlation between the type of sacral agenesis and a specific urodynamic pattern. It is concluded that virtually all children with an anorectal malformation and associated sacral agenesis have neurological bladder-sphincter dysfunction and therefore should receive early urodynamic assessment and appropriate urological treatment. PMID- 8708914 TI - Viral IL-10 gene therapy inhibits TNF-alpha and IL-1 beta, not IL-6, in the newborn endotoxemic mouse. AB - INTRODUCTION: Modulation of the inflammatory cascade within the liver of critically ill infants may improve the chance of survival. Using gene therapy, the authors hypothesized that augmented local production of the counter regulatory cytokine viral interleukin-10 (IL-10) in vivo will modulate the critical cytokines in the inflammatory response. The purpose of the present study was to determine whether replication-defective adenovirus-mediated viral IL-10 (vIL-10) gene transfer and expression within the liver can achieve this goal in newborn mice. MATERIALS AND METHODS: Four-week-old Balb/c mice were administered (intraperitoneally) 1 x 10(9) plaque-forming units (pfu) per milliliter of an adenovirus vector (E1a/b-deleted) than encodes the sv40 promoter and the BCRF1 cDNA, or of control vector dl434 that expresses no foreign gene. Forty-eight hours later the mice were challenged with 50 micrograms/kg of lipopolysaccharide (LPS) they were killed 1, 2, 6, or 24 hours later (six at each time point). Southern blot analysis was performed on genomic DNA isolated from the liver, lung, and kidney to assess gene transfer of BCRF1. Homogenized liver protein was analyzed for tumor necrosis factor-alpha (TNF-alpha), IL-1 beta, IL-6, and recombinant vIL-10. RESULTS: Southern blot analysis confirmed successful gene transfer to the liver but not to the lung, kidney, or dl434-transduced liver in mice that received adenovectors. Viral IL-10 levels within the liver ranged from 14 to 18 ng/mL. In controls, TNF-alpha production was elevated at early time points, to 18,000 pg/mL, but decreased rapidly by 24 hours after LPS challenge. The TNF-alpha levels of animals treated with Ad5svBCRF1 were significantly lower than those of controls throughout the course of study (P < .0001). After the LPS challenge, hepatic IL-1 beta decreased, from a maximum of 800 pg/mL (2 hours) to 411 pg/mL (24 hours). Inhibition of IL-1 beta by vIL-10 occurred at 1 hour (P > .016) and 2 hours (P < .001) only. Hepatic production of IL-6 after LPS challenge ranged from 7 to 8,000 pg/mL in all groups and was not altered by vIL-10 gene therapy. CONCLUSION: In vivo administration of adenovectors encoding BCRF1 to newborn mice results in efficient hepatic transduction and expression of recombinant vIL-10. The Kupffer cell response to LPS is suppressed with respect to TNF-alpha and IL-1 beta, but not IL-6. In vivo modulation of hepatic cytokine responses is achievable using gene products that mimic cellular cytokines. This is an effective model for the selective evaluation of therapeutic gene products for gene therapy of sepsis. PMID- 8708915 TI - Partial intestinal obstruction induces substantial mucosal proliferation in the pig. AB - Parenteral nutrition and improving supportive treatment have resulted in prolonged survival for patients with short bowel syndrome. However, definitive therapy for patients with short bowel syndrome must focus on increasing small intestinal mucosal mass. Intestinal lengthening procedures rely on intestinal dilation to accomplish this. The authors hypothesized that partial intestinal obstruction would result in consistent dilation of the intestine and would provide increased intestinal mass for use in intestinal lengthening. The authors developed a partially obstructing prosthetic valve to dilate the intestine before intestinal lengthening. This report describes the changes elicited by the valve. Twelve weanling pigs were divided randomly into two groups of six. One group had valve placement 240 cm distal to the ligament of Treitz; the other had sham surgery. The survival rate was 100% for both groups, and the mean weight gain was similar. Both groups were fed pig chow mush and were killed 5 weeks after surgery. Intestinal diameter was measured, and the small intestine was harvested, preserved, and sectioned for microscopic examination. The mean bowel diameter 15 cm proximal to the valve was 4.7 cm in the valved group and 3.3 cm in the sham group (42% increase). Total mucosal thickness, villus height, crypt depth, and villus density were significantly greater for the valved pigs in all sections (proximal and distal to the valve). Surface index and intestinal circumference were significantly greater in the valved pigs in all sections proximal to the valve, but there was no significant difference in these values for sections distal to the valve. There was no significant difference in villus cell density between the two groups at any location. Chronic partial obstruction of the small intestine results in consistent dilation of the intestine, with growth of all layers of the bowel, including the mucosa. This dilation and mucosal growth results in a true increase in surface area and is an ideal first step toward sequential lengthening. PMID- 8708916 TI - The structure of the transitional and aganglionic zones of Auerbach's plexus in patients with Hirschsprung's disease: a computer-assisted three-dimensional reconstruction study. AB - Intestines resected from two patients with Hirschsprung's disease, both aged 6 months, were submitted to serial sectioning and three-dimensional reconstruction to visualize the overall structure of Auerbach's plexus and correlate its changes with functional abnormalities. Reconstruction was made possible by using a graphics computer system. In normal intestines taken from an autopsy case, both large and small, the plexus was shown extending along the intermuscular septum as a regular network. But in the large intestine, the density of the network was apparently higher and the individual bundles thicker than in the small intestines. This neural network was absent in the aganglionic zone in Hirschsprung's disease, as expected, where only hypertrophic extrinsic nerves were running in the septum. The "transitional zone" of Hirschsprung's disease was clearly definable as an area extending over a certain length, where the network meshes grow more irregular and the bundles taper more, toward the aganglionic zone. This finding was considered to be of profound significance in the design of a surgical strategy to prevent postoperative bowel dysfunction. In determining the range of resection, one must consider not only the presence or absence of nerve cells but also the grade of plexus hypoplasia. PMID- 8708917 TI - Entrapment of the appendix and the fallopian tube in peritoneal dialysis catheters in two children. AB - Continuous ambulatory peritoneal dialysis (CAPD) has revolutionized the management of renal failure in children. However, CAPD catheter malfunction is not uncommon, the main complications being catheter-related infection and catheter-related obstruction. Herein the authors report both of the above complications, attributable to unique and previously unreported causes. One patient had appendicitis and peritonitis secondary to catheter entrapment of the appendix; the other had an obstructed catheter secondary to entrapment of the right fallopian tube within its lumen. To avoid potentially serious complications, early exploration of malfunctioning catheters is recommended. PMID- 8708918 TI - Conn syndrome in a child, caused by adrenal adenoma. AB - Hyperaldosteronism owing to aldosterone-producing adenoma (Conn syndrome) is a rare but potentially curable form of pediatric hypertension. The authors report on a 5-year-old girl who had symptoms of polyuria, polydipsia, and fatigue, and for whom the diagnosis of hyperaldosteronemia was suggested by a low serum potassium level and persistent hypertension. The diagnosis was confirmed by increased levels of plasma aldosterone and decreased levels of plasma renin. The tumor was localized with ultrasonography and computed tomography, which showed a 2-cm mass in the left adrenal gland. The left adrenal gland was excised, and pathological assessment showed an adenoma. Only 14 other pediatric cases (< 16 years of age) have been reported in the English-language literature. PMID- 8708919 TI - Microphthalmos associated with esophageal atresia. AB - The authors report on a neonate who had esophageal atresia and tracheoesophageal fistula associated with unilateral microphthalmos and glandular hypospadias. Primary repair of the esophageal defect was accomplished in the first week of life. This is the first report of such a combination of congenital anomalies. PMID- 8708920 TI - Hepatogonadal fusion. AB - Fusion of the testis with adrenal remnants is a relatively common incidental finding during orchidopexy. Splenogonadal fusion has been described in several cases. Herein the authors report a case of hepatogonadal fusion. To their knowledge, this is the first such case in the literature. Ectopic liver tissue has been encountered in a variety of locations, but had not been found in a testicle. The trapping of hepatocyte-destined mesenchyma in different areas may explain the presence of heterotopic liver tissue in different organs. If these cell aggregates are trapped in the area of peritoneum where the testis is forming, heterotopic liver tissue can end up in the testis. PMID- 8708921 TI - Primary hyperparathyroidism in infancy: a case report. AB - Primary hyperparathyroidism is a rare disease of childhood. The condition is even rarer in the neonatal and infant stages. The disease, with its main manifestation hypercalcemia-often is fatal. The authors successfully treated a 2.5-month-old boy who had primary parathyroid hyperplasia. The patient had recurrent pneumonia and failure to thrive. Blood test results showed an abnormally high level of calcium, which was resistant to medical therapy. Further investigations showed high levels of parathyroid hormone. The patient underwent neck exploration, which showed hyperplasia of the all four parathyroid glands. Total parathyroidectomy was performed, with one gland being autotransplanted to the deltoid muscle. The patient had an immediate hypocalcemic period, followed by normocalcemia. In light of the present case and others in the Literature, the authors recommended total parathyroidectomy followed by autotransplantation of a gland to an accessible muscle. PMID- 8708922 TI - Endoscopic laser division of an esophageal web in a child. AB - The authors report the first successful endoscopic laser division of a congenital esophageal web in a child in whom dilatation had failed. Laser lysis is minimally invasive and should be attempted before surgery is considered. PMID- 8708923 TI - Congenital megalourethra. AB - The authors report on two patients who had fusiform megalourethra with unusual features. The first patient had left hydronephrosis with duplication of the ureter, oblique sagittal septum in the urinary bladder, and Y duplication of urethra with megalourethra of both channels. The second had features of the abdominal muscle deficiency syndrome, with anterior ectopic anus, rectourethral fistula, and fusiform megalourethra. Both patients were treated by preliminary urinary diversion. The first patient subsequently had staged reconstruction. The other patient died of urinary sepsis. The authors believe that preliminary urinary diversion followed by planned staged reconstruction offers hope for the reconstruction and rehabilitation of these cases. PMID- 8708924 TI - Central-type eight-toed polydactyly associated with ipsilateral complex renogluteal agenesis: a case report with 8 years' follow-up. AB - At birth, a baby girl was found to have central-type eight-toed polydactyly of the left foot associated with ipsilateral nonvisualization of the kidney and atrophy of the gluteal maximum muscle. X-ray examination showed eight completely developed metatarsal and digit bones, and a chromosome study showed a normal 46, XX karyotype. She received ray amputation of the extra toes when she was 1 year old. Follow-up examination at age 9 showed unchanged renogluteal anomalies and a slightly smaller left foot. In addition, a bony defect over the posterior iliac spine and a wide femoral neck-shaft angle were found on the radiograph. Her left leg was found to be 1 cm shorter than the right, but there was no evidence of valgus or varus. Presently she can walk and run quite well, and there is no callus formation over her sole. The timing and options for excision of the supernumerary digits are discussed and this rare disease reviewed. PMID- 8708925 TI - A case of recurrent pulmonary hypertension in a 41-gestational-week newborn. PMID- 8708926 TI - Prenatal hormonal therapy improves pulmonary compliance in the nitrofen-induced CDH rat model. PMID- 8708927 TI - Why has conventional intrapartum cardiotocography not given the expected results? AB - It is probable that conventional electronic fetal monitoring (EFM) has reduced the intrapartum death rate, but the expected dramatic reduction in neurological handicap has not occurred. There are two reasons for this: the majority of infants, who develop neurological problems have been harmed before the onset of labor, and the method of EFM has been more difficult to use in daily routine than expected. However, EFM is the best method we have to monitor high risk cases and the results can be improved by better training of the staff. PMID- 8708928 TI - Improvements in computerized fetal heart rate analysis antepartum. AB - The continued development of a computerised system for measuring the pattern of the antepartum fetal heart rate (FHR) is described. Previous work had established that measurement of FHR variation objectively detects chronic fetal hypoxaemia and the onset of metabolic acidaemia antepartum. The normal centiles were calculated for the amplitude of long-term FHR variation, in episodes of high and low variation, week by week from 24-42 weeks gestation. Reference to these (automatically by the computer) improved discrimination between normal and questionable records in 38% of records, with a small saving of time. Two types of sinusoidal rhythm were described (slow, 1 in 2-5 minutes, incidence 0.16% of subjects; and faster, 2-5 per minute, incidence 0.025%) with methods for their detection. Both may be of sufficient amplitude to induce an episode of high FHR variation. The different effects of maternal steroid (betamethasone or dexamethasone) administration of FHR variation were compared, and the clinical consequences considered. The frequency distribution of basal FHR in normal and abnormal records was measured, and the effects on basal FHR outside the normal range (120-160 bpm) on FHR variation described. Adjustment of the FHR baseline was undertaken when, exceptionally, large abrupt changes in heart rate occurred during a record. The duration and frequency of FHR record acquisition in clinical practice were reviewed, and new policies recommended. With adequate safeguards, measurement by a computer offers reliable objective information from which fetal health may be assessed, more objectively and accurately than by visual inspection. PMID- 8708929 TI - Fetal monitoring by combined CTG and FBA. AB - Cardiotocography still is the main method to monitor the unborn child during birth. Today, problems occurring during CTG-surveillance are not so much due to the method, but a consequence of insufficient management. Fetal monitoring by combined CTG and FBA brings advantages for mother and child. The use of FBA in cases with pathological fetal heart patterns leads to a significant reduction in cesarean section rate, neonatal morbidity and perinatal mortality. PMID- 8708930 TI - Antenatal assessment using the FECG obtained via abdominal electrodes. AB - During the past decade a variety of intrapartum fetal monitors have been constructed that process the entire fetal electrocardiogram (FECG), obtained via a scalp electrode. They therefore differ from conventional monitors in aiming to extract relevant timing and magnitude information from the morphology of the FECG rather than simply the RR interval and hence heart rate. An intrapartum monitor such as this has been successfully developed by ourselves. This paper describes the early results obtained whilst attempting to extend this form of monitoring forward into the antenatal period. In order to achieve this the FECG must be acquired via surface electrodes placed on the maternal abdomen, which yields a signal containing the FECG amidst a number of noise sources. Our investigations into the feasibility of "antenatal abdominal FECG analysis" have been on two fronts. The first has been to produce a bedside monitor similar in function to our intrapartum device, whilst the second has been to address the possibility of performing such monitoring in ambulant subjects. At present the antenatal bedside monitor has successfully extracted and processed the FECG in approximately 75% of the cases studied, with subjects ranging from 20 weeks through to term having been monitored. We also have demonstrated the feasibility of the long term monitoring of maternal and fetal heart rate using a portable instrument. PMID- 8708931 TI - Continuous fetal monitoring of HbO2 with pulse oximetry. AB - Pulse oximetry is well established in anesthesia and intensive care medicine. Application for fetal monitoring would be desirable, but significant modifications are necessary. We report our first promising clinical experience with a self-developed probe, hard- and software system. We hope by the positive results which we have observed that this new technology--oxicardiotocography (OCTG) will improve the safety of fetal monitoring. PMID- 8708932 TI - Cerebral tissue oxygenation during hypoxia and hyperoxia using artificial placentation in lamb. AB - Aiming at a better understanding of the pathophysiologic basis of perinatal encephalopathy, we evaluated patterns of tissue oxygenation during hypoxia and hyperoxia. We utilized both laserspectroscopy and invasive tissue-Po2 microneed measurements synchronously in five newborn lambs (141-143 days of gestation). The model of artificial placentation provided defined changes of the blood gases, using a extracorporeal circuit with interposition of membrane lung. During hyperoxia, the Po2 at the blood outlet port of the lung was raised to > 300 mmHg for five minutes. During hypoxia, Po2 was diminished as oxygen at the gas phasis was replaced by nitrogen. After the induction of hyperoxia, a rise of tissue-Po2 was observed. The synchronously recorded data of the laserspectroscopy showed adequately rising HbO2 values in concordance (r = 0.97, p < 0.001). As a constant finding we did not observe Cyt-aa3 changes during induced hyperoxia with tissue Po2 values of < 40 mmHg. Furthermore, no changes in blood volume occurred in this case. A different pattern of the laserspectroscopic parameters was found when the tissue-Po2 rose above a value of > 40 mmHg and Cyt-aa3 rose after a lag-time occurred. During induced hypoxia an immediate fall of tissue-Po2 corresponding with a fall of HbO2 in the spectroscopic tracing occurred (r = 0.87, p < 0.001). A fall of the Cyt-aa3 level was seen with a lag-time when the tissue-Po2 had reached values of below 10 mmHg. In addition, a rise of blood volume was recorded in all cases of induced hypoxia. In conclusion, the results indicated that cellular redoxe state remains stable over a large range of oxygen partial pressure changes. PMID- 8708933 TI - Correlation between NIR spectroscopy and pulse oximetry in the fetus. AB - At the Women's University Hospital Wurzburg, pulse oximetry as well as Near Infrared Spectroscopy (NIR) measurements are performed on the fetus during labor in order to compare both methods. Changes of oxygen saturation seem to be positively correlated to concentration changes of oxidized hemoglobin measured by NIR, partially correlated to those of total hemoglobin, and partially correlated negatively to those of reduced hemoglobin. No correlation was found between oxygen saturation changes and those of cytochrome aa3, perhaps due to a longer lagtime. Pulse oximetry as well as NIR Spectroscopy may prove helpful tools for fetal surveillance. Both must be studied particularly with reference to FBA and fetal outcome before clinical decisions can be influenced by these methods. PMID- 8708934 TI - Comments on past and present situation of intensive monitoring of the fetus during labor. AB - Modern intensive monitoring of the fetus during labor affords the opportunity for the greatest possible safety to the infant, whilst at the same time dispensing with unnecessary operations to terminate labor which can be dangerous for the mother and her child. Developments over the past 28 years, since cardiotocography has been in clinical use, have unfortunately also been accompanied by confusion and mistakes. We think that there are two main reasons for this: I. Many who use cardiotocography have not assessed its efficiency correctly. Heart rate pattern with all their variations have too often been regarded as the only diagnostic criteria for the determination of fetal hypoxia. However this procedure only allows a rough selection of actually endangered fetuses. In more than 50% of the cases with a suspicious or pathologic cardiotocogram there is no concrete clinical evidence of manifest danger of hypoxia. II. Investigations concerning the practical importance of monitoring the fetus during labor were often not based on actual clinical indications. Attempts were made to prove that continuous electronic monitoring of the fetal heart rate could be replaced by simple traditional auscultation. Furthermore due to failure to appreciate the real efficiency of cardiotocography, this truly excellent method was in fact blamed for the faults arising from wrong interpretation, instead of underlining the lack of logic of the investigators concerned. After all, several prospective studies, that have examined the efficiency of various monitoring strategies, demonstrate the shortcomings of these techniques in the clinical setting and have shown them to have no relevance in every practice. Presently the most suitable way to monitor the fetus during labor is to combine cardiotocography with a biochemical method. At the moment fetal blood analysis is the most suitable test to be performed in cases where the cardiotocogram is not normal. Then the role of fetal blood analysis is to find out whether in fact a danger of hypoxia is threatened or not. Today the most urgent task when monitoring the fetus during labor is not so much to reduce the number of cases of cerebral palsy since these are seldom caused by intrapartum asphyxia. The major objective is now recognised to be to prevent early morbidity of the fetus, newborn, and the consequences that flow from this. The excellent results of combined intensive monitoring of the fetus during labor are particularly well illustrated by the intrapartum mortality in our hospital during different periods when monitoring practice has been progressively and methodically improved by stages. During the period when monitoring consisted of simple auscultation used on its own, the intrapartum mortality was 0.56%. After the introduction of fetal blood analysis--at that time cardiotocography did not yet exist--the mortality sank to 0.32%. During the first ten years of cardiotocography combined with fetal blood analysis the mortality reduced further to 0.17%, and during the following 13 years it reached to a minimal level of 0.4%. Finally, in this review, other aspects are commented on such as oximetry. Oximetry, presently used as pulse oximetry, as an additional biochemical method will possibly be incorporated as the third method of monitoring the fetus during labor. Oximetry could also be used as an automatic alarm system in the labor ward when fetal O2 levels drop below a particular limit to draw the attention of the staff to the case concerned. Up to now this has not been convincingly achieved with the use of cardiotocography. PMID- 8708935 TI - Relationship between amniotic fluid insulin and maternal blood glucose concentrations in patients with carbohydrate intolerance during pregnancy. AB - In contrast to maternal blood glucose, amniotic fluid insulin (AFI) directly reflects the functional state of the fetal pancreas. In a prospective study we evaluated the correlation of AFI with maternal metabolic control in 70 amniotic fluid specimens from 61 women having carbohydrate intolerance during pregnancy (White A n = 44, B0 n = 17). AFI was measured with the Insulin RIA 100 kit from Pharmacia (Freiburg). The normal range of AFI was established in 304 healthy pregnant women (16th-42nd gestational week). AFI concentrations increased by a factor of 1.5 to 2 during gestation reflecting the maturation of the fetal pancreas. Elevated AFI levels (> 97th centile) were found in 11% of normoglycemic diabetics and in 50% of women with insufficient metabolic control. Despite a high overall concordance (81%) no direct relationship could be found between fetal and maternal parameters. Patients with increased AFI values had a 5-fold higher rate of large-for-gestational age (LGA) infants than women with normal levels. This finding confirms the pathogenetic role of hyperinsulinism in the development of fetal macrosomia. PMID- 8708936 TI - Biochemical monitoring of fetal distress with serum-immunoreactive erythropoietin. AB - The ability of parameters like umbilical arterial pH and Apgar score to predict and/or to reflect fetal distress are limited. It is known that erythropoietin (EPO) increases when partial pressure of oxygen is insufficient for metabolic demand. Therefore we studied the levels of EPO in the cord blood of stressed neonates (n = 75). In addition, reference values for EPO were established in a group of healthy term infants (n = 54) (mean +/- SD: 20.02 +/- [mU/ml]) and in premature infants (n = 77) according to gestational age. A significant increase in EPO concentrations was found in the stressed group: 153.4 +/- 418.8 [mU/ml], p < 0.003 (n = 27) in acute stress; and 102.6 +/- 127.1 [mU/ml], p < 0.002 (n = 48) in chronic stress. However parameters like hemoglobin, hematocrit, umbilical arterial pH and Apgar-score did not correlate with EPO values. A sensitivity of 59% and a specificity of 92% was calculated. We conclude that serum EPO concentrations are capable of detecting acute and chronic stress and could be useful as a screening method. In part EPO concentrations also allow us to grade stress in pregnancies that are complicated by diseases like preeclampsia. PMID- 8708937 TI - Basal layer of epithelium expresses tenascin mRNA during healing of incisional skin wounds. AB - Tenascin is a large oligomeric glycoprotein of the extracellular matrix that is expressed prominently during embryonic development and wound healing. Previous studies on tenascin expression in wounds have used immunohistochemistry to describe the expression of tenascin in wounds. The present study used in situ hybridization to identify the cells expressing tenascin mRNA in healing wounds. The results demonstrate that the cells of the basal layer of epidermis, migrating over the healing wound, are expressing the mRNA for tenascin. Intense expression was seen during the first three days after wounding, but after seven days, after the epithelium had grown to cover the wound, no tenascin transcripts were seen in epithelial cells. The epithelial cells elsewhere in the skin were devoid of tenascin transcripts at all stages examined. Previously, prominent immunohistological staining for tenascin has been located in wounds below the migrating epithelial cells and it has been thought to be synthesized by stromal cells upon epithelial induction. Our findings in the present study indicate that tenascin is produced by epithelial cells, which apparently are induced to produce tenascin as they migrate after wounding. PMID- 8708938 TI - Histamine H1 receptor-stimulated Ca2+ signaling pathway in human periodontal ligament cells. AB - We studied histamine-induced Ca2+ mobilization in human periodontal ligament (HPDL) cells. Histamine induced a transient rise in intracellular Ca2+ ([Ca2+]i) and maintained a sustained phase in the presence of extracellular Ca2+. In the absence of extracellular Ca2+, the transient peak was slightly reduced and the sustained phase was decreased to the basal level. The initial rise in [Ca2+]i was attributed to two components: intracellular Ca2+ release and Ca2+ influx, whereas the sustained phase was due to Ca2+ influx. After depletion of intracellular Ca2+ stores with thapsigargin, a known Ca(2+)-ATPase inhibitor, histamine-induced increase in [Ca2+]i was significantly reduced, suggesting histamine induces Ca2+ release from inositol 1,4,5-trisphosphate [Ins(1,4,5)P3]- and thapsigargin sensitive Ca2+ stores. Histamine-induced peak in [Ca2+]i was increased dose dependently in the presence and absence of extracellular Ca2+. The histamine mediated response in [Ca2+]i was specifically attenuated by chlorpheniramine (H1 antagonist) but not by cimetidine (H2 antagonist), clearly indicating that activation of H1 receptor mediates histamine-induced Ca2+ mobilization. We next examined the effect of histamine on inositol phosphates formation. Histamine stimulated the formation of inositol phosphates which changed time-dependently. In particular, the formation of Ins(1,4,5)P3 was increased significantly for 10 s. The histamine-induced Ca2+ mobilization caused an increase of prostaglandin E2 (PGE2) release which was reduced in excluding extracellular Ca2+. These results indicate that activation of histamine H1 receptor induces the accumulation of Ins(1,4,5)P3 and the following transient increase in [Ca2+]i, and elicits the release of PGE2 which may be coupled with Ca2+ influx. PMID- 8708939 TI - Comparison of the pro-inflammatory cytokine-stimulating activity of the surface associated proteins of periodontopathic bacteria. AB - Saline extraction of the periodontopathic bacterium, Actinobacillus actinomycetemcomitans, releases surface-associated material (SAM), a complex mixture of proteins and carbohydrates with potent biological actions on isolated bone and on various mammalian cell populations. In this study, the relative ability of the SAM from 5 organisms, implicated in the pathology of periodontal disease, to stimulate human mesenchymal and myelomonocytic cells to synthesize the proinflammatory cytokines - interleukin (IL)-1 beta, IL-6 and tumour necrosis factor (TNF)alpha has been investigated. The bacteria investigated were Actinobacillus actinomycetemcomitans, Eikenella corrodens, Porphyromonas gingivalis, Prevotella intermedia and Campylobacter rectus. Human cells were exposed to a four log order range of concentrations of the SAM, or of Escherichia coli lipopolysaccharide, to provide full agonist dose responses in order to allow comparison of the potency and efficacy of each SAM. All SAMs demonstrated the capacity to stimulate human gingival fibroblasts (HGFs), human peripheral blood mononuclear cells (PBMCs) or the myelomonocytic cell line - Mono-Mac-6 to release one or all of the cytokines assayed. Activity was heat- and trypsin-sensitive suggesting that the active components were proteinaceous. However, there were substantial differences in the potency and efficacy of each SAM when compared on a concentration basis (w/v). The most active SAM was from A. actinomycetemcomitans with those from E. corrodens and P. gingivalis being slightly less active. The least active cytokine-stimulating SAMs were from C. rectus and Pr. intermedia. One major difference between the SAMs and E. coli LPS was the inability of the former to stimulate HGFs to release IL-1 beta or TNF alpha although they could stimulate PBMCs to release these cytokines. This may have relevance to the pathology of the periodontal diseases. PMID- 8708940 TI - Evidence that cyclosporine inhibits periodontal prostaglandin I2 synthesis. AB - Cyclosporine (CsA) is a selective immunosuppressant widely used in clinical therapy. Like phenytoin and nifedipine, the drug is associated with gingival overgrowth. This study considers the interaction of CsA and prostaglandin I2 (PGI2), in particular the action of the drug on gingival tissue in vitro and in vivo. The PGI2-synthesis of rat, rabbit and human gingival tissue was examined by bioassay. In vivo CsA-therapy reduces gingival PGI2-synthesis. The results furthermore show a dose-dependent inhibition of PGI2-synthesis by CsA (1-100 micrograms/ml) in vitro. PGI2-synthesis from in vivo CsA-pretreated probes was further dose-dependently diminished by in vitro addition of CsA. As PGI2 exerts an antiproliferative activity via cAMP-elevation, the drug-induced inhibition of PGI2 production is claimed to be responsible for gingival hyperplasia in CsA treated patients. PMID- 8708941 TI - Detection of stable and active periodontitis sites by clinical assessment and gingival crevicular acute-phase protein levels. AB - The aim of the present study was to investigate whether incipient periodontal disease breakdown could be associated with changes in gingival crevicular fluid (GCF) acute-phase protein levels. In addition, the potential of clinical indices to act as predictors of significant attachment level (AL) change was investigated. AL measurements were taken at baseline and 3 months using the Florida Probe stent handpiece from a total of 384 sites in 38 patients. The average standard deviation of duplicate AL measurements was 0.423. When the tolerance method was used to detect significant AL change, 3.9% of the sites lost attachment. When a less stringent criterion of AL change of > or = 1 mm was used 9.9% of the sites lost attachment during the 3-month period. With the exception of probing depth, baseline clinical parameters failed to predict AL change. Fourteen active periodontitis sites that demonstrated significant attachment loss were paired to stable periodontitis sites within the same patient. The levels of four acute-phase proteins, namely alpha 2-macroglobulin (alpha 2-M), alpha 1 antitrypsin (alpha 1-AT), transferrin (TF) and lactoferrin (LF), and also albumin (Alb) were assessed in the same gingival crevicular fluid sample using sandwich ELISAs. Results were expressed either as ng/30 s and ng/microgram Alb. Acute phase protein levels in GCF failed to differentiate between active and stable periodontitis sites at baseline. In conclusion, the degree of gingival inflammation of the tissues adjacent to the crevice/pocket seems to influence the levels of protease inhibitors and iron-binding proteins in GCF to a greater extent than probing attachment loss. PMID- 8708942 TI - Human cementum protein extract promotes chondrogenesis and mineralization in mesenchymal cells. PMID- 8708943 TI - Fluoride distribution in human dental calculus obtained from different sites on the tooth surface. AB - We examined the site specificity of fluoride (F) distribution in human dental calculus. Teeth with supra- and subgingival calculus were obtained from patients who resided in non-fluoridated areas in Japan and China. Sequential layers of the dental calculus (30 microns thick) were abraded by an abrasive micro-sampling technique and fluoride and phosphorus in the powdered samples were analyzed. Fluoride concentrations were highest in the outer, lowest in the middle and intermediate in the inner layers of dental calculus in general. In the outermost layers fluoride concentrations were highest in calculus found near the tooth cervix both in supra- and subgingival calculus. Fluoride concentrations decreased markedly toward the apical region in subgingival calculus, while it did not change toward the incisal or occlusal region in supragingival calculus. In the inner layers, fluoride concentrations in both supra- and subgingival calculus were not affected by position on the teeth. Fluoride concentrations in subgingival calculus near the apex were lower than in supragingival calculus near the incisal or occlusal region. It was concluded that the fluoride concentrations differ in different regions of dental calculus, probably due to their different mechanisms of formation. PMID- 8708944 TI - Influence of retinoic acid on the expression of cytokeratins, vimentin and ICAM-1 in human gingival epithelia in vitro. AB - Phenotypic differences exist in vivo between junctional (JE) and oral gingival (OGE) epithelia and an in vitro system has been developed that maintains phenotypic differences. This system, which permits in vitro studies of factors that may influence the epithelial phenotype, was used to investigate the effects of retinoic acid (RA) on epithelial expression of various markers known to distinguish JE from OGE. Primary cultures of JE and OGE were initiated from defined gingival regions and were subcultured and grown for 48 h in 96-well plates or on multiple-well slides. Control cultures were grown in medium supplemented with delipidized serum and all-trans RA was added to experimental groups. Other cultures were grown in a defined RA-free medium. Cultures were examined using monoclonal antibodies against cytokeratins, vimentin, and ICAM-1 and binding displayed by indirect immunocytochemical staining. Staining reactions were assessed by direct microscopic observation and assayed by spectrophotometric quantitation. The results showed that RA had minor effects on the marker expression of JE but markedly enhanced expression of cytokeratins 8, 18, 19, vimentin and ICAM-1 in OGE. These markers, which normally distinguish JE from OGE, were expressed at levels approaching or exceeding those of control JE cultures. These observations indicate that RA responsive mechanisms affect the phenotypes expressed by epithelia in vitro and suggest that such mechanisms may be related to the different phenotypic patterns expressed by gingival epithelia in vivo. PMID- 8708945 TI - Production of inflammatory mediators and cytokines by human gingival fibroblasts following bacterial challenge. AB - Bacteria can indirectly affect the course of periodontal diseases by activating host cells to produce and release inflammatory mediators and cytokines. These mediators and cytokines manifest potent proinflammatory and catabolic activity and may play key roles in local amplification of the immune response as well as in periodontal tissue breakdown. This study tested the effect of Actinobacillus actinomycetemcomitans (Aa) and Campylobacter rectus (Cr) challenge on PGE2, IL-1 beta, IL-6 and IL-8 production by human gingival fibroblasts (HGF). Contact inhibited HGF were prepared and formalin-killed bacterial cells (Aa JP2, ATCC 29523 & 33384 and Cr ATCC 33238) at 10(6)-10(9) were added to the HGF. Culture supernatants were collected at varying time intervals and analyzed for cytokine and mediator content. All concentrations of Aa JP2 and Cr ATCC 33238 suppressed IL-1 beta production up to approximately 50% during the initial 3-12-h period. No bacterial concentration tested was able to increase IL-1 beta production above the maximum basal levels. Both bacterial species stimulated production of IL-6 and IL-8. Aa JP2 did not affect PGE2 levels significantly, whereas Cr ATCC 33238 was stimulatory only at the highest concentration tested (10(9)). There were no significant differences among the three Aa strains with respect to IL-1 beta production. However, Aa ATCC 29523 and ATCC 33384 were less capable of stimulating IL-6 secretion and more efficient in stimulating IL-8 production than Aa JP2. In general, Cr was the most potent enhancer of cytokine and mediator production by HGF. In conclusion, Aa and Cr are capable of amplifying the local immune response and promoting periodontal tissue inflammation by stimulating HGF to secrete mainly IL-6 and IL-8. PMID- 8708946 TI - Nicotine effects on PGE2 and IL-1 beta release by LPS-treated human monocytes. AB - Cigarette smoking is a major risk factor in the development and further progression of periodontitis. However, little is known regarding the pathogenesis of smoking-related periodontal diseases. The purpose of this study was to examine the effects of nicotine, alone and in combination with lipopolysaccharide (LPS), on monocyte secretion of bone-resorbing factors, PGE2 and IL-1 beta. Peripheral blood monocytes (PBM) were isolated by counterflow centrifugal elutriation from 15 healthy, non-smoking donors. PBM were incubated for 24 h in RPMI 1640 containing nicotine (0, 50 ng/ml, 1 microgram/ml, 10 micrograms/ml and 100 micrograms/ml) with or without 10 micrograms/ml Porphyromonas gingivalis LPS or Escherichia coli LPS. Culture supernatants were assayed for PGE2 and IL-1 beta by ELISA. None of the nicotine preparations resulted in significant PBM secretion of PGE2 and IL-1 beta above that of unstimulated cultures. However, PGE2 release was potentiated 1.7-fold by the combination of P. gingivalis LPS and 10 micrograms/ml nicotine relative to P. gingivalis LPS alone (p < 0.05, one-way ANOVA). Prostaglandin E2 release also was potentiated 3.5-fold by P. gingivalis LPS and 100 micrograms/ml nicotine relative to P. gingivalis LPS alone (p < 0.00001, one way ANOVA) and 3.1-fold by E. coli LPS and 100 micrograms/ml nicotine relative to E. coli LPS alone (p < 0.00001, one-way ANOVA). IL-1 beta secretion was lower for either LPS plus 100 micrograms/ml nicotine relative to LPS alone, although not significantly. These data demonstrate upregulation of LPS-mediated monocyte secretion of PGE2 by nicotine and suggest a potential role for nicotine in periodontal disease pathogenesis. PMID- 8708947 TI - Use of chlorhexidine chewing gum significantly reduces dental plaque formation compared to use of similar xylitol and sorbitol products. AB - A double-blind 3-treatment crossover design employing a 6-day trial period with out mechanical oral hygiene measures was used to compare dental plaque formation following use of chlorhexidine (CHX) acetate-, xylitol-, and sorbitol-containing chewing gum. Fourteen dental students were assigned a random ordering of the chewing gum products and received professional tooth cleaning at the start of trial periods. For each trial period, subjects were instructed to use 5 pieces of the unlabeled chewing gum daily (containing 5.0 mg CHX acetate/piece; 0.8 xylitol/piece; or 1.0 g sorbitol/piece). Two pieces of chewing gum were used following each morning and evening meal and one piece following the noon meal. The subjects were instructed to use the products for 20 minutes at each occasion. A 7-day washout-period between trial periods was used. The Turesky modification of the Quigley and Hein index was used to assess plaque formation. Differences between treatments were evaluated using a repeated measures ANOVA with Newman Keuls multiple comparisons. The CHX-containing chewing gum showed significantly reduced plaque values (0.7 +/- 0.4) compared to the sorbitol-(2.7 +/- 0.4; P < 0.01) and xylitol-product (1.7 +/- 0.3; P < 0.01). Furthermore, the CHX-product significantly reduced plaque levels compared to the study subjects' regular plaque control routines (1.3 +/- 0.04; P < 0.05). The xylitol-product exhibited significantly lower plaque-values than the sorbitol-product (P < 0.01). Our results suggest that regular use of CHX-containing chewing gum appears useful to control dental plaque formation. PMID- 8708948 TI - In vitro comparison of aged and young osteogenic and hemopoietic bone marrow stem cells and their derivative colonies. AB - The purpose of this in vitro study was to determine whether there were differences in the number and size of osteogenic and hemopoietic colonies derived from bone marrow stem cells of aged and young adult male Sprague-Dawley rats. Using a Ficoll-Paque gradient, stem cells were harvested from aged male rats 18 to 22 months old and young adult males 55 days of age. Single cell suspensions from the red marrow of the long bones were cultured 14 days in vitro and subsequent colonies were assessed by light microscopy for number and size. A computerized histomorphometric linear measuring system was utilized to assess colony area in square millimeters. The results clearly show that young animals have a statistically significant increased cellular potential for osteogenic and hemopoietic colony formation. Cultures from aged animals showed an average formation of 0.45 +/- 0.6863 osteogenic colonies while those from younger animals had an average of 3.6 +/- 2.3523 osteogenic colonies per 3 million cells plated. Hemopoietic colonies from aged animal cell cultures numbered 5.25 +/- 2.2449 while those from the young animals averaged 8.23 +/- 3.3601 per 3 million cells plated. The difference in size of the osteogenic and hemopoietic colonies between age groups was not statistically significant. The area of osteogenic colonies derived from aged animals measured 0.1244 +/- 0.0891 mm2, while those derived from the young animals averaged 0.1276 +/- 0.0518 mm2. Hemopoietic colonies from the aged cells measured 0.0759 +/- 0.0514 mm2, while hemopoietic colonies from the young animal cells measured 0.06010 +/- 0.0180 mm2. The results of this study may have implications for consideration in the cellular healing aspects of aged versus young individuals. PMID- 8708949 TI - Influence of tooth instrumentation roughness on gingival tissue reactions. AB - Histological studies have demonstrated a relationship between the amount of subgingival plaque and the magnitude and extension of gingival tissue reactions. The objective of the present study was to evaluate inflammatory reactions in the gingival tissues facing plaque accumulation at a diamond and curet-instrumented root surfaces. Experimental, deep periodontal defects were established at buccal surfaces of mandibular and maxillary canine teeth in 5 beagle dogs. The root surfaces were instrumented by a flame-shaped, fine-grained. rotating diamond point, or by a sharp curet. Next, the dogs were fed a plaque-inducing diet for 70 days. The animals were then sacrificed, and tissue blocks of the experimental sites including teeth, alveolar bone, and gingival tissues were secured. The gingival soft tissue was processed for histomorphometric analyses at 3 levels. Epithelium and connective tissue area measurements showed no differences between the two instrumentations. Junctional epithelium (JE) cell point counts exhibited a higher proportion of inflammatory cells (IC)in specimens facing diamond compared to curet-instrumented defects. A higher proportion of IC was present within the coronal compared to the apical aspect of the JE for both instrumentations (P < 0.05). A significant difference in IC density between instrumentations was detected for non-infiltrated (P < 0.05), as well as for infiltrated (P < 0.01) connective tissue. The infiltrated connective tissue (ICT) inflammatory cell density was significantly (P < 0.01) and positively correlated to the JE inflammatory cell density (r = 0.75), and to area measurements of ICT (r = 0.55). The overall results demonstrate that the character of subgingival root instrumentations significantly affects gingival inflammatory reactions, most likely by influencing subgingival plaque formation. PMID- 8708950 TI - Effect of periodontal treatment on the protein composition of whole and parotid saliva. AB - Cystatins are physiological inhibitors of cysteine proteinases and widely distributed in human tissues and fluids including saliva. Cystatins S, SA, SN, and D are only found in glandular saliva and tear fluid whereas cystatin C has been detected in all tested biological fluids. Previous studies demonstrated that total cystatin activity and cystatin C concentration were highest in whole and parotid saliva of periodontitis patients compared to healthy subjects suggesting a response of the salivary glands to an inflammatory condition of the oral cavity. Based on these results we studied a possible change in the concentration of cystatin S, cystatin C, albumin, IgA, amylase activity, and cystatin activity in a whole and parotid saliva of 20 periodontitis patients as a consequence of periodontal treatment. Saliva samples were quantified for cystatins S and C, albumin, and IgA by enzyme-linked immunosorbent assay. Amylase was determined in an activity assay and total cystatin activity was measured towards pa-pain. The clinical condition of the subjects improved significantly after 6 months of periodontal therapy whereas biochemical analyses of whole and parotid saliva indicated that significant changes in salivary protein composition occurred only in whole saliva. Total cystatin activity (P < 0.05) and cystatin C concentration (P < 0.05) of whole saliva samples collected after periodontal treatment decreased to normal healthy control values. Further, concentrations of cystatin S were unchanged during the periodontal treatment process. These results suggest that other sources of cystatins than the parotid gland i.e.; other salivary glands or crevicular fluid, are involved in the decrease of total cystatin activity in whole saliva after periodontal treatment. PMID- 8708951 TI - Compliance with supportive periodontal therapy. AB - Analysis of 1,280 private patient records from a periodontal clinic showed poor compliance with supportive periodontal therapy (SPT). Data were analyzed according to sex, age, and type of treatment (with or without periodontal surgery). Analysis showed that 25.2% of the patients never returned and among those who report for SPT, only 40.1% did so regularly; the percent of surgical cases was greater (70.7%) and those patients had better compliance (77.6%); compliance of women was greater (76.5%); abandonment of supportive periodontal treatment was high (66.7%), and compliance increased with age. Several suggestions concerning clinical procedures are offered with the objective of motivating patients to give the necessary importance to supportive periodontal therapy. PMID- 8708952 TI - Periodontal regeneration of human intrabony defects with bioresorbable membranes. A controlled clinical trial. AB - The purpose of this controlled clinical trial was to compare the clinical efficacy of 3 treatment modalities in the treatment of deep interproximal intrabony defects. Thirty-six (36) defects in 36 patients were randomly assigned to 1 of 3 treatment groups by blocking to prognostic variables. The test was treated with bioresorbable membranes positioned coronal to the interproximal bone crest; the second group (membrane control) was treated with conventional non resorbable (ePTFE) barrier membranes applied coronal to the alveolar crest; the third group (flap Control) was treated with an access flap procedure (MWF). No differences were observed in terms of baseline oral hygiene and defect characteristics among the 3 groups, indicating that the blocking approach was effective. A stringent infection control program was enforced for 1 year. The results indicated that: 1) at 1 year all treatment modalities resulted in clinically significant improvements in clinical attachment levels (CAL) and reductions in probing depths; 2) a statistically significant treatment effect (P < 0.0001, ANOVA) was observed comparing the test (4.6 +/- 1.2 mm), the membrane control (5.2 +/- 1.4 mm), and the flap control groups (2.3 +/- 0.8 mm) in terms of CAL gain; 3) differences in terms of CAL gain between the test (bioresorbable) and the membrane control (ePTFE) groups were not statistically significant (P = 0.19, t-test); 4) both the test and the membrane control groups gained significantly more CAL at 1 year than the MWF group (P < 0.0001, t-test). CAL gains > or = 4 mm were observed in 83.3% of cases in both GTR groups, while CAL gains of this magnitude were not detected in the MWF group. We concluded that clinically significant CAL gains can be obtained with GTR procedures using both bioresorbable and non-resorbable membranes. Patients' morbidity, however, was lower in the group treated with bioresorbable membranes. PMID- 8708953 TI - Prostaglandin E2 secretion, cell maturation, and CD14 expression by monocyte derived macrophages from localized juvenile periodontitis patients. AB - Previous studies from our laboratory have demonstrated that freshly isolated peripheral blood monocytes from localized juvenile periodontitis (LJP) patients secrete more prostaglandin E2 (PGE2) after stimulation by lipopolysaccharide (LPS) than do monocytes from healthy subjects. However, it is not clear if the altered function of LJP monocytes is intrinsic to the cells or is induced by the persistent infection of the periodontium. The present study was designed to compare PGE2 secretion by freshly-isolated peripheral blood monocytes (FIM) from LJP and control subjects to in vitro monocyte-derived macrophages (MDM) from the same subjects. We also examined monocyte maturation into macrophage-like cells and the cell-surface expression of the LPS, receptor, CD14 during the culture period. FIM from LJP patients and controls were stimulated by different concentrations of LPS (O to 30 micrograms/ml) for 24 hours. These experiments were performed immediately after cell separation and after 10 days in culture, which allowed differentiation of monocytes into MDM. PGE2 levels in the culture media were determined using a radioimmunoassay. Cell surface expression of CD71, a cell maturation marker, and CD14 were assayed by cell-ELISA in relation to beta 2-microglobulin. LPS-stimulated FIM from LJP patients secreted 3 to 4 times more PGE2 than control FIM at all LPS concentrations tested. After 10 days in culture, the LJP MDM secretion of PGE2 reduced to control MDM level of PGE2 secretion. These levels of PGE2 secretion were comparable to PGE2 secretion from FIM of controls. Cell maturation, as verified by CD71 expression, was found not to differ between the groups. However, the expression of CD14 by LJP FIM was much lower than on control FIM (approximately equal to 50%). After 5 or 10 days in culture, MDM from both control and LJP subjects expressed comparable amounts of CD14. The results suggest that in vitro conditions reverse the hypersensitivity of LJP monocytes to LPS into control levels and CD14 expression is not correlated to the hyper-responsiveness of the cells to LPS. PMID- 8708954 TI - Effects of various hygiene procedures on the surface characteristics of titanium abutments. AB - The use of cleaning instruments on titanium implants may cause undesired surface alterations. In a qualitative and quantitative assessment of these alterations, 5 titanium implant abutments were treated with a steel curet, a prototype pure titanium curet, an air abrasive polishing system, and an ultrasonic system. Custom-made polymer templates, used to secure the curet to a vertical guide bar and a spring scale to maintain a constant instrument pressure, guaranteed a standardized procedure and reproducible results. The ultrasonic and the air abrasive polishing method were also standardized. Evaluation by scanning electron microscopy (SEM) revealed surface alterations for all instruments and systems except the plastic curet, which did not roughen the surface at all. The confocal laser-scanning microscope allows a 3-dimensional reproduction of these surface alterations and their direct measurement. The profilometric tracing was not sensitive enough to register the minor effects caused by the titanium curet and the air abrasive polishing system. Dimensions of the resulting surface microstructure could be determined with the laser-scanning microscope. Since the influence of such surface defects on the peri-implant tissue reaction is unpredictable, the titanium curet and the air abrasive system can only be recommended with restrictions. The steel curet and the ultrasonic system proved to be totally unsuitable for cleaning titanium implants. PMID- 8708955 TI - Migration of human gingival fibroblasts over guided tissue regeneration barrier materials. AB - The purpose of this investigation was to determine the ability of human gingival fibroblasts, in vitro, to migrate along a chemotactic gradient over 3 different guided tissue regeneration barrier materials; i.e., polytetrafluoroethylene, polylactic acid, and sterile calcium sulfate. Forty petri dishes were divided into 4 equal groups. In each group of 10 dishes, a different barrier material served as the fibroblast substrate with the polystyrene floor of one group of Petri dishes serving as the control. The under agarose technique of measuring cell migration was employed using platelet derived growth factor-BB homodimer as the chemoattractant and Hanks balanced salt solution to test random migration. In addition, fibroblasts were directly cultured on triplicate sets of barrier materials and the control surface for 24 hours and examined by scanning electron microscopy. Comparative analysis of the fibroblast migration data showed the mean migration distance (adjusted for random migration) for controls to be significantly greater than any of the three barrier materials. Further, mean migration distance over calcium sulfate was significantly greater when compared to that of the polylactic acid barrier group. All other comparisons between groups were not statistically significant. Scanning electron microscopic examination fibroblasts cultured directly on barrier membranes and compared to controls indicated that the calcium sulfate substrate appeared to facilitate cell attachment and spreading whereas cells on polytetrafluoroethylene and polylactic acid barriers exhibited a morphology not conducive to migration or, in many cases, cell health. Based on these limited in vitro results and, given the 3 barrier materials considered, it would appear that calcium sulfate offers the greater potential for guided tissue regeneration in surgical sites where primary wound closure cannot be obtained. PMID- 8708956 TI - Clinical and microbiological observations of early polytetrafluoroethylene membrane exposure in guided bone regeneration. Case reports in primates. AB - Premature membrane exposure at one week is described in 3 Macaca mulatta monkeys as part of a sequence of experiments on guided bone regeneration. Clinical sequelae include redness, edema, and tissue slough. Bacteroides fragilis, Streptococcus pneumoniae, Prevotella intermedia, and Staphylococcus intermedius were detected at all prematurely exposed sites. Pseudomonas maltophilia, Strept, pneumoniae, and P. intermedia were the predominant organisms detected and consisted of more than 10% of the total anaerobic count. PMID- 8708957 TI - Development of an exostosis following a free gingival graft: case report. PMID- 8708958 TI - Immunological, genetic, and microbiological study of family members manifesting early-onset periodontitis. AB - From the standpoint of host-parasite interactions, family studies help us understand the host defensive factors and the molecular mechanisms involved in the periodontal immune response. In this study, we report the immunological profile of host-defensive functions, human leukocyte antigen (HLA) phenotypes, and the microflora of a mother (rapidly progressive periodontitis), an older son (periodontally healthy), a younger son (localized juvenile periodontitis), and a daughter (localized juvenile periodontitis). We examined the peripheral neutrophil functions, phenotypic and functional analysis of peripheral lymphocytes, serum immunoglobulin G (IgG) antibody titers against periodontopathic bacteria, serological type of HLA class II antigens, and bacterial flora in all periodontal pockets. The results showed that Actinobacillus actinomycetemcomitans was dominant in the pockets of all subjects. The mother and two sons showed a depressed neutrophil chemotaxis to N-formyl methionyl-leucyl-phenylalanine. All subjects except the older son exhibited low T4/T8 ratios. The mother and daughter had raised levels of IgG titers to Porphyromonas gingivalis. All subjects had HLA phenotypes of DRw52 and DQ1 in common. We found that the family members had similar disorders in certain defensive functions. This family has been a model for our understanding of the host defensive factors in the development of early-onset periodontitis. PMID- 8708959 TI - PDGF-B producing cells and PDGF-B gene expression in normal gingival and cyclosporine A-induced gingival overgrowth. AB - It has been proposed that healthy gingiva is in a continuous state of wound repair. Thus, one might expect to find cells in normal gingiva producing growth factors associated with wound healing such as platelet-derived growth factor B chain (PDGF-B). One might also expect to find increased numbers of these cells or increased amounts of these growth factors in conditions which involve increased tissue volume such as drug-induced gingival overgrowth (DGO). The purpose of this study was to quantify PDGF-B gene expression and identify cells producing PDGF-B in normal gingiva and DGO. Cyclosporine A (CSA) was selected as a prototype of the overgrowth condition. Twelve patients with clinical CSA DGO and 12 patients with no DGO or history of drugs known to cause DGO were selected for study. Frozen sections of gingival specimens from these patients were subjected to in situ hybridization for PDGF-B mRNA. Positive cells were counted and expressed as mean +/- SEM cells/mm2 of lamina propria. Morphometric analysis revealed 6.2 +/- 1.9 cells/mm2 for control gingiva and 10.3 +/- 3.4 cells/mm2 for CSA DGO samples. There was no statistically significant difference between groups. PDGF-B gene expression was measured in these cells and expressed as mean +/- SEM silver grains/cells. There was a significant upregulation of PDGF-B gene expression in cells from the CSA DGO group (39.5 +/- 14.7 silver grains/cell for normal gingiva vs. 255.3 +/- 77.1 silver grains/cell for CSA DGO samples; P < 0.001). The presence of PDGF-B in these cells was confirmed in all cases by immunocytochemical localization. Additionally, PDGF-B producing cells were identified as macrophages in sections taken from an additional patient with CSA DGO by double immunofluorescence labeling of the CD51 membrane marker for macrophages and intracellular PDGF-B. These findings are consistent with the concept that healthy gingiva is in a continuous state of wound repair and support the hypothesis that CSA DGO is associated with enhanced macrophage PDGF-B gene expression rather than an increase in the number of PDGF-B producing macrophages. PMID- 8708961 TI - Effectiveness of subgingival scaling and root planing: single versus multiple episodes of instrumentation. AB - This study evaluates the effectiveness of subgingival scaling and root planing comparing the effect of a single instrumentation to the effect of three instrumentations. A total of 35 teeth in 15 patients were selected; 15 were scaled once (Group A), 15 were scaled three times (Group B), and 5 were used as controls (Group C), representing teeth that were not instrumented. The Group A and B teeth were chosen in the same patient based on random selection. All the teeth were scored by the calculus index of the periodontal disease index. Six surface locations were probed to determine probing depth. The level of the gingival margin was marked on the teeth to locate supra- and subgingival calculus after extraction. The Group A and B teeth received the initial episode of scaling and root planing for not more than 10 minutes, then only the Group B teeth received two additional instrumentations of not more than 5 minutes each. The additional instrumentations were performed 24 hours after the initial scaling. The scaled and control teeth were extracted immediately after the third instrumentation period. The teeth were washed with water and stained with methylene blue. They were viewed under a stereomicroscope which had a tenth grid on its eyepiece. Assessments were made involving the total counts and percents of the surfaces covered with calculus on the scaled and unscaled teeth. The results demonstrated no significant difference in the effectiveness of calculus removal between single and multiple episodes of scaling and root planing. Similar results were found for the total amount of calculus removed, the calculus removed from individual surfaces, and the calculus removed from various probing depth levels. PMID- 8708960 TI - Cyclosporine A upregulates platelet-derived growth factor B chain in hyperplastic human gingiva. AB - Cyclosporine A (CSA) is a widely used immunosuppressant for transplant patients and is also used for the treatment of a wide variety of systemic diseases with immunologic components. A prominent side effect of CSA administration is gingival overgrowth (hyperplasia). It has been postulated that CSA alters fibroblast activity through effects on various growth factors/cytokines. However, as yet, data concerning the molecular mechanisms involved in pathologic connective tissue proliferation are preliminary in nature. Our previous investigations concerning phenytoin-induced effects on platelet-derived growth factor B (PDGF-B) gene expression have demonstrated that other drugs which cause gingival overgrowth can upregulate macrophage PDGF-B gene expression in vitro and in vivo. The purpose of the present study was to evaluate PDGF-B gene expression in gingival tissues of patients receiving CSA therapy and exhibiting gingival overgrowth to determine if similar PDGF-B upregulation occurs in response to CSA and to identify PDGF-B producing cells in these tissues. Quantitative competitive reverse transcription polymerase chain reaction (QC-RTPCR) techniques were utilized to measure PDGF-B mRNA levels in CSA overgrowth patients and normal controls (N = 6/group). Results were expressed as mean +/- mRNA copy number and tested for significance using unpaired t-tests. Gingival samples were harvested (standardized for local inflammation at the sample site), total RNA was extracted, and QC-RTPCR was performed using specific PDGF-B primers and a corresponding competitive internal standard. CSA-treated patients exhibiting gingival overgrowth demonstrated approximately 48-fold increase in PDGF-B mRNA (7667.1 +/- 477.4 copies for CSA patients vs. 158.2 +/- 37.1 copies for controls; P < 0.001). Additionally, dual fluorescence immunohistochemistry for mature macrophage marker antigen (CD51) and intracellular PDGF-B was utilized to identify and localize PDGF-B producing cells were demonstrated to be macrophages distributed in a non-uniform manner throughout the papillary connective tissue. These results further support the hypothesis that the molecular mechanisms responsible for drug-induced gingival overgrowth may involve upregulation of PDGF-B macrophage gene expression. We continue to investigated specific CSA-induced alterations of macrophage PDGF-B gene expression in vitro and in vivo. PMID- 8708962 TI - Engineering specifics of the periodontal curet's cutting edge. AB - Much has been reported on the qualitative, and to some degree, the quantitative effects of wear and sharpening of the periodontal curet's cutting edge. Presented here is an engineering model of the periodontal curet's cutting edge during wear and sharpening, and an estimated life of a typical periodontal curet. Although not confirmed by scientific experiments, this engineering model provides a framework for further scientific analysis. It also gives a clinical appreciation for the longevity of such periodontal instruments. PMID- 8708963 TI - Radiographic evaluation of bone regeneration following periodontal surgery with or without expanded polytetrafluoroethylene barriers. AB - In 16 patients with advanced periodontitis, 23 teeth exhibiting interproximal intrabony defects were treated by conventional periodontal surgery (n = 10, control) or guided tissue regeneration (GTR) technique using expanded polytetrafluoroethylene (ePTFE) barriers (n = 13, test), respectively. Clinical parameters were assessed before and 6 months after surgery. Presurgically and 3 and 6 months postsurgically standardized bite-wing radiographs were taken. Using a loupe and a computer-assisted system, respectively, the distances from the cemento-enamel junction (CEJ) to alveolar crest (AC) and CEJ to the most apical extension of bony defect (BD) were measured. The average gain of attachment was assessed 2.33 mm in the control group and 3.17 mm in the test group. The bony fill was measured 0.97 mm and 0.97 mm in the control group and 0.93 mm and 1.68 mm in the test group after 3 and 6 months, respectively, using a loupe. Using a computer-assisted system 0.83 mm and 1.82 mm of bone fill could be measured (control) and 0.76 mm and 1.79 mm (test) after 3 and 6 months, respectively. Compared to the gold standard of surgical measurements, the computer-assisted analysis of radiographs underestimated bone loss significantly less than evaluation with a loupe (P < 0.002). Compared to conventional periodontal surgery, the GTR technique resulted in higher attachment gain and bony fill. However, the high variation of surgical results in the test group prevented the calculation of a statistically significant difference. PMID- 8708964 TI - Clinical significance of furcation anatomy of the maxillary first premolar: a biometric study on extracted teeth. AB - Multirooted teeth in which chronic periodontitis has progressed to involve the furcation present special problems in treatment. The apical placement of furcation, narrow furcation entrance, concavities on the root surface, and the undesirable root configuration make instrumentation and plaque control difficult. Furcation involvement, when present in maxillary first premolars in particular, presents a poor prognosis as the furcation opens mesiodistally. This study on 100 extracted maxillary first premolars is an attempt to determine the frequency of bifurcation, the proximity of bifurcation to cemento-enamel junction, the depth of concavities on the root trunk and the furcal aspect of root, the diameter of furcation entrance, and the extent of root divergence. PMID- 8708965 TI - Alveolar ridge preservation following extraction of maxillary anterior teeth. Report on 23 consecutive cases. AB - The purpose of this article was to clinically evaluate the predictability of a surgical procedure to preserve the alveolar ridge after extracting maxillary anterior teeth with advanced bone loss. This procedure is indicated when a tooth supported fixed partial denture will rehabilitate the area. Non-resorbable hydroxyapatite crystals are used as graft material and a rotated pediculated split thickness palatal flap covers them. Twenty-three patients who were treated with this procedure were followed for 12 to 24 months. Results were predictable with minimal postoperative ridge deformation. Indications for this procedure are discussed. PMID- 8708966 TI - A new monitor with a zinc-oxide thin film semiconductor sensor for the measurement of volatile sulfur compounds in mouth air. AB - Halitosis, defined as unpleasant oral odor, is a concern among the general public. Halitosis is generally diagnosed by organoleptic examination and by gas chromatographic analysis of the main source of halitosis, volatile sulfur compounds, such as hydrogen sulfide, methyl mercaptan, and dimethyl sulfide. Gas chromatography requires a large-scale system and a long running time. We investigated the use of a zinc-oxide thin film semiconductor sensor for measuring trace volatile sulfur compounds in mouth air. Mouth air samples collected in teflon bags from 21 volunteers were analyzed by 3 methods: the monitor analysis, gas chromatography, and organoleptic examination by 3 judges. The readings of the monitor were correlated with the values of the total volatile sulfur compounds measured by gas chromatography (r = 0.75, P < 0.01) and also with the organoleptic scores given by the judges (r = 0.76, P < 0.01). The organoleptic scores were correlated with the gas chromatographic values (r = 0.71, P < 0.01). These results suggest that this new monitor with a zinc-oxide thin film semiconductor sensor may be used for the diagnosis of halitosis. Its small size and simplicity of handling may enable its use for routine chair-side study and field surveys of halitosis. PMID- 8708967 TI - Repair following treatment of circumferential periodontal defects in dogs with collagen and expanded polytetrafluoroethylene barrier membranes. AB - This study was designed to evaluate healing following treatment of periodontal defects using 2 collagen barrier membranes with different degrees of cross linking, and to compare the results to those following use of an expanded polytetrafluoroethylene (ePTFE) membrane. Horizontal, circumferential defects were created in mandibular premolar teeth of 6 beagle dogs, followed by placement of membranes and wound closure. Postoperative healing was monitored clinically for 6 months, at which time the animals were sacrificed and specimens were taken for histological examination. Clinical observations indicated that the highly cross-linked, slow-resorbing collagen membrane did not integrate with the tissues the way the less crosslinked, rapid-resorbing collagen did. Membrane exposure was typical for the slow-resorbing membrane in contrast to the rapid-resorbing membrane which remained covered. The inferiority of the slow-resorbing membrane was evident by the extensive clinical recession and the attachment level measurements taken at 6 months, and it was decided to omit this membrane from histometric analysis. Histological examination of root surfaces treated with rapid-resorbing collagen or ePTFE membranes revealed substantial reparative healing. The connective tissue repair amounted to 84% of the treated root surface height for the rapid-resorbing collagen and 53% for the ePTFE membrane (difference not statistically significant). However, the connective tissue repair to the rapid-resorbing collagen group root surfaces was often associated with a layer of ankylosis (44%, versus 8% of the ePTFE group). It appeared that much of the ankylotic response was initiated from the furcation area and extended around to the buccal and lingual surfaces. In addition, this study provided histological evidence that granulation tissue forming under clinically exposed and plaque contaminated ePTFE membranes can still result in connective tissue repair. It was concluded that the rapid-resorbing collagen membranes and the ePTFE membranes seem capable of stimulating periodontal connective tissue repair, whereas the slow-resorbing collagen membranes were unsuccessful in this effort, as seen in this dog model. PMID- 8708968 TI - Osteoclast inhibition by factors from cells associated with regenerative tissue. AB - Guided tissue regeneration (GTR) uses expanded polytetrafluoroethylene (ePTFE) membranes to favor the repopulation of the healing wound with cells with bone regenerative potential. As bone remodeling is a tightly coupled process, inhibition of osteoclast-mediated bone resorption may be critical to regeneration. Thus, this study was undertaken to determine whether cells associated with regenerative tissue can inhibit osteoclast differentiation and activity and to begin characterizing and identifying the factor(s) mediating the observed effects. Conditioned media were harvested from human periodontal cell lines established in culture: cells adherent to ePTFE membranes, recovered from patients after GTR; cells adherent to ePTFE augmentation membranes, recovered from edentulous ridge augmentation procedures; and periodontal ligament cells from periodontally healthy bicuspids. Conditioned medium from each of these regenerative cell lines reduced the number of tartrate-resistant acid phosphatase positive osteoclast-like cells formed from hemopoietic precursors in mouse bone marrow cultures. Also, both the total resorbed surface area and number of resorption pits formed by these cells on calcium phosphate ceramic films were reduced. The factor in the conditioned medium which inhibited osteoclast differentiation was soluble, heat labile, and resided in the lower molecular weight (< 30 kDa) fraction, the same fraction which would contain cytokines. Western blot analysis of the conditioned medium detected a band at the molecular weight of interferon gamma (IFN-gamma), using a polyclonal rabbit anti-human IFN gamma. Thus, the factor in the conditioned medium with inhibitory activity may have identity with IFN-gamma or one of the other anti-inflammatory cytokines. PMID- 8708969 TI - Microbial flora in the acute phase of periodontitis and the effect of local administration of minocycline. AB - Periodontitis, similar to other infectious diseases, is known to progress as chronic inflammation with recurrent acute phases. The purpose of this study was to clarify the microbiological composition of the acute phase and to compare the bacterial flora with that of comparable chronic periodontal pockets. We also evaluated the effect of application of minocycline gel locally on the change in the microflora in the acute pockets. Microbial flora from the subgingival pockets of 28 patients in the acute phase of periodontitis and of 12 patients in a comparable chronic phase as the control were investigated by various bacterial culture methods including TS blood agar and TSBV plates. Minocycline gel was applied to the acute periodontal pockets. Changes in the microbiological proportion and clinical parameters at one week after baseline examination were followed by dark-field analysis, culture method, and indirect immunofluorescence technique. Characteristic features of bacterial proportions in the acute site were observed as an increase in Bacteroides forsythus. The number of Porphyromonas gingivalis and black pigmented anaerobic rods also increased. Application of minocycline gel in the acute pocket without any debridement produced improvement in clinical symptoms at one week. Black-pigmented anaerobic rods, P. gingivalis, and B. forsythus decreased significantly at one week after the application. Results indicate that periodontopathic bacteria including B. forsythus and P. gingivalis were predominant in the acute phase of periodontitis and a locally delivered antibiotic may be effective as an alternative modality of treating the acute inflammation. PMID- 8708970 TI - Effect of selected beta-blockers on supragingival calculus formation. AB - Supragingival plaque and calculus indices, salivary flow rates, pH, ionic and total calcium concentrations, total phosphate concentration, and the number of cervical restorations and caries lesions were measured in 29 subjects using systemic beta-(beta) blockers and in 28 subjects who were not taking any systemic medication. After 8 weeks of normal oral hygiene following an oral prophylaxis, a second comparison of each of the above quantities was made. With the exception of the calculus indices and the incidence of cervical restorations, no significant differences were found for any of the measured quantities between the medicated and non-medicated groups at either examination. The medicated group showed significantly lower mean calculus values than the non-medicated group at both examinations and a higher incidence of cervical restorations at the baseline examination, suggesting that beta-blockers decrease the rate of mineralization in the oral cavity. Since beta-blockers did not appear to alter stimulated salivary pH, flow rate, phosphate, ionic calcium or total calcium concentrations, their effect on the mineralization processes must be attributed to other mechanisms. Two hypotheses appear plausible: changes in salivary mineralization rates caused by either direct physico-chemical effects of the secreted beta-blockers in the saliva, or by alterations in the salivary protein/glycoprotein composition, enzymes and oral bacterial flora owing to systemic pharmacological effects of beta-blockers. PMID- 8708971 TI - Host defensive functions in a family manifesting early-onset periodontitis. AB - Family case studies help us identify host risk factors in periodontal disease. In this study we examine a family consisting of a mother (40 years old, with rapidly progressive periodontitis), her elder daughter (14 years old, with localized juvenile periodontitis), and younger daughter (13 years old, with simple gingivitis). We examined 1) the peripheral neutrophil functions (chemotactic migration, phagocytosis, superoxide production); 2) lymphocyte functions (proliferative activity and cytokine productivity of T cells, immunoglobulin [Ig] M productivity of B cells when stimulated with pokeweed mitogen); 3) phenotypic analyses of peripheral lymphocyte subpopulations; 4) serum IgG antibody titers against periodontopathic bacteria; and 5) serological type of HLA class II. All the subjects exhibited high T4/T8 ratios due to high percentage of CD4-positive cells, showed high IgG titers to Actinobacillus actinomycetemcomitans, and had a HLA DQw1 in common. The mother showed a slight deficiency of neutrophil chemotactic migration to N-formyl methyonyl leucyl phenylalanin (fMLP), raised interleukin-2 productivity of T cell, and high levels of IgG titers to Porphyromonus gingivalis and Fusobacterium nucleatum. Both daughters showed weak T cell proliferative response to anti-CD3 monoclonal antibody and low IgM productivity. Low lymphocyte responsiveness may be involved in the pathogenesis of periodontal disease of these daughters; therefore, the lymphocyte dysfunctions shown should be considered in relation to the progression of periodontal disease. PMID- 8708972 TI - Adjacent gingival recession treated with expanded polytetrafluoroethylene membranes: a report of 2 cases. AB - We report two cases illustrating treatment for adjacent facial gingival recession using guided tissue regeneration (GTR) with expanded polytetrafluoroethylene membranes. The clinical results showed that GTR obtained soft tissue coverage of the root surfaces and was effective in reducing hypersensitivity and improving the esthetics. In both cases, satisfying clinical results have been maintained for 6 to 12 months after re-entry procedure. PMID- 8708973 TI - Juvenile hyaline fibromatosis of gingiva: a case report. AB - Juvenile hyaline fibromatosis is an extremely rare inherited condition, probably resulting from an inborn error of metabolism. It is characterized by cutaneous nodules, gingival hypertrophy and joint contractions. It affects children but usually it is not present at birth, and is microscopically characterized by a conspicuous hyalinization of the connective tissue. PMID- 8708975 TI - The place of scientific publications in the evaluation of a clinical success. PMID- 8708974 TI - The importance of regular dental treatment in patients with cyclic neutropenia. Follow-up of 2 cases. AB - Data on two patients with cyclic neutropenia are presented. They demonstrate that regular tooth care and professional dental treatment can prevent progressive periodontal breakdown but that neglecting oral hygiene soon leads to periodontal pathology. Regular, monthly professional removal of dental plaque and calculus, and rinsing with 0.2% chlorhexidine gluconate during neutropenia help maintain periodontal attachment level. The caries susceptibility and the apical periodontitis in the intact anterior tooth of the female patient indicate the possibility of cyclic neutropenia playing a role in caries and pulpal pathology. PMID- 8708976 TI - Community dwelling elderly are appropriate subjects for intensive dietary choice restriction studies. AB - There is a traditional belief that the elderly have difficulty coping with dietary change, and therefore have a diminished likelihood of successfully responding to nutritional interventions or restrictions. Using a controlled mild zinc-deficiency feeding study as a model for strict dietary intervention, we assessed psychological responses to severe dietary choice restriction in 15 Caucasian, elderly (66.12 +/- 4.43 years) males (n= 7) and females (n = 8). Participants completed the Medical Outcomes Study Short Form 36-Item Questionnaire (SF-36) as an index of QOL and the Multi-dimensional Health Locus of Control (MHLC) as a measure of health beliefs at pre-intervention baseline, post-intervention, and follow-up. No subjects dropped out nor were any meals missed during the entire 21-day feeding study period. No significant differences were detected across time on the MHLC (Internal F = 0.53, P = 0.6; Powerful Others F = 0.28, P = 0.8; Chance F = 1.1, P = 0.4.) by one-way ANOVA. Similarly, for the SF-36 no significant differences were found across time (F = 0.76, P = 0.5). Our results suggest that restricting dietary choices does not negatively impact older adult subjects and that they can cope well with dietary choice restriction and change. Older adults should not be overlooked for nutritional intervention solely due to age considerations. PMID- 8708977 TI - Using the Nutrition Screening Initiative to survey the nutritional status of clients participating in a home delivered meals program. AB - A large percentage of older Americans are at risk for malnutrition. This puts them at risk for premature institutionalization, creating a financial burden. The objective of this survey was to determine the nutritional health of clients receiving home delivered meals in Lake County, Indiana and the impact that home delivered meals had on them. Nutrition Screening Initiative (NSI) Determine Your Nutritional Health Checklists were mailed to recipients of meals; 58.3% were returned. Twenty-eight percent were found to be at no nutritional risk, 39% at moderate nutritional risk and 33% at high nutritional risk. One-hundred-thirty clients that scored three or more on the "Checklist" were visited by a Registered Dietitian for further screening using the NSI Level I Screen. This screen found many nutritional problems but the fact that the clients did receive home delivered meals decreased the risk. It was determined by the author that 68% of these clients could not function in their own homes without home delivered meals. PMID- 8708978 TI - Appropriate protocol for zinc therapy in long term care facilities. PMID- 8708979 TI - Effect of alginate and alginate-cimetidine combination therapy on stimulated postprandial gastro-oesophageal reflux. AB - This randomized, single-blind cross-over study compared the effectiveness of a conventional alginate reflux barrier formulation (20 mL single dose of Liquid Gaviscon; sodium alginate, sodium bicarbonate, calcium carbonate) with a 20 mL single dose of an alginate-cimetidine combination formulation (Algitec Suspension; sodium alginate, cimetidine) in the suppression of food and acid reflux into the oesophagus after a test meal in 12 healthy volunteers. Subjects were fasted overnight before the study. A pH electrode and gamma detector were accurately positioned 5 cm above the cardia. The volunteers received a 99mTc labelled meal designed to provoke reflux and then either remained untreated, or 30 min later were given either Algitec Suspension or Liquid Gaviscon. Reflux of both food and acid into the oesophagus was measured for 3 h. There was a seven day wash-out period between each treatment. Food reflux in the control group was 22,878 +/- 14,385 counts x 10(3) and this was significantly suppressed by both Liquid Gaviscon (174 +/- 128 (s.e.) counts x 10(3); P = 0.003); however, although the reduction of food reflux to 3812 +/- 2322 counts x 10(3) observed after Algitec treatment was considerable, this did not reach statistical significance (P > 0.05) due to the large intersubject variation. Liquid Gaviscon was significantly better at reducing food reflux than Algitec (P = 0.001). Gaviscon also significantly reduced acid reflux when compared with the control group (1.08 +/- 0.73 vs 5.87 +/- 3.27% recording time oesophageal pH < 4, respectively) (P = 0.03). The slight reduction in acid reflux after Algitec treatment (3.25 +/- 1.82% recording time oesophageal pH < 4) also did not reach statistical significance. The difference between Algitec and Gaviscon treatment was also not significant. PMID- 8708980 TI - Prediction of cathodal iontophoretic transport of various anions across excised skin from different vehicles using conductivity measurements. AB - Solute concentration, buffer concentration, applied pH and buffer constituents affect the cathodal iontophoresis of salicylate, benzoate and butyrate across excised skin. Experiments were conducted in which the iontophoretic flux of salicylate was measured across excised human skin with variations in salicylate concentration, donor solution pH, buffer concentrations and buffer consitutents. The conductivity of these solutes and of solutions described in studies on the iontophoresis of benzoate and butyrate were then measured. The observed variations in salicylate, benzoate and butyrate fluxes across a range of conditions were found to be related to the ratio of specific conductance of the solutes in deionized distilled water to that in the buffer solution containing the solutes. The present results suggest that conductivity of solutes in vehicle solutions is one means of predicting the iontophoretic flux of solutes from different vehicle compositions. PMID- 8708981 TI - Percutaneous absorption of salicylates from some commercially available topical products containing methyl salicylate or salicylate salts in rats. AB - Studies to determine the extent of local tissue penetration of topically applied, commercially available salicylate esters and salts were conducted in male Wistar rats. The salicylate concentration in plasma, tissues underlying the site of drug application, and similar tissues on the contralateral (control) side were measured. The plasma and tissue salicylate levels suggest that direct penetration of salicylate was predominant to the top muscle level on the treated site. Results also suggest that the drugs were first absorbed into the bloodstream and subsequently distributed to both the deeper tissues on the treated site and the contralateral tissues. The topical application of formulations of ester methyl salicylate and salts triethanolamine salicylate and diethylamine salicylate containing comparable salicylate concentrations yielded similar salicylate concentrations in the various tissues. The salicylate concentrations in the deeper tissues approached concentrations observed in the contralateral tissues suggesting that salicylate present in these tissues was due to the systemic blood supply. PMID- 8708982 TI - Effect of 2-hydroxypropyl-beta-cyclodextrin on percutaneous absorption of methyl paraben. AB - A potential use of 2-hydroxypropyl-beta-cyclodextrin (HP-beta-CyD) to solubilize methyl paraben and to suppress its percutaneous absorption was examined, and compared with nonionic surfactant HCO-60. HP-beta-CyD significantly increased the solubility of methyl paraben in water, where the apparent 1:1 stability constant of the soluble complex was determined to be 2150 M-1. The in-vitro cutaneous permeability of methyl paraben through an isolated skin of hairless mouse was suppressed by HP-beta-CyD, thus promoting the bioconversion of methyl paraben to the less toxic metabolite, p-hydroxybenzoic acid (p-HBA) in the epidermis. These effects of HP-beta-CyD were greater than those of HCO-60. HP-beta-CyD (2% w/v) reduced the in-vivo percutaneous absorption of methyl paraben by 66% 24 h after the topical application of a solution containing [14C]methyl paraben to hairless mouse skin. Additionally, the percutaneous absorption of [14C]HP-beta-CyD was confirmed to be extremely low. These results suggest that HP-beta-CyD is useful in liquid preparations of methyl paraben for topical application. PMID- 8708984 TI - Aggravating process induced by indomethacin on chronic gastric lesion in rat. Role of polymorphonuclear leucocytes. AB - The present study examines the indomethacin-provoked aggravation of chronic ulceration induced by acetic acid in rats. The drug was administered in a single dose, 7 and 14 days after provocation of ulceration. The changes induced by indomethacin in other groups of animals that had been treated for 7 and 14 days with hydroxyurea (which provokes a marked leucopenia) were also studied. The results obtained demonstrate that indomethacin does not significantly modify the macroscopic index of ulceration nor vascular permeability in the majority of the groups tested. Only in the group that received hydroxyurea for 14 days was there an increase in both parameters. Myeloperoxidase activity was assayed and used as an index of leucocyte infiltration in an attempt to relate the increase in this activity with a gastrolesive effect. Application of acetic acid produced a significant increase in this activity 7 days after induction of chronic injury. Administration of hydroxyurea intraperitoneally was associated with a decrease in the severity of chronic ulceration and neutrophil infiltration into the gastric mucosa. This effect was detectable enzymatically and microscopically. The groups that received indomethacin showed an increase in myeloperoxidase activity, although this increase was only significant in the animals treated with hydroxyurea for 7 and 14 days. The results suggest that the aggravation provoked by indomethacin is greater when the ulcer curing process is more advanced. PMID- 8708983 TI - Intragastric distribution of ion-exchange resins: a drug delivery system for the topical treatment of the gastric mucosa. AB - Previous studies by this group on freeze-dried oral dosage forms containing finely-divided ion-exchange resins revealed prolonged gastric residence and uniform distribution within the stomach. The present study was carried out to ascertain whether this was due to freeze-drying, properties of the radiolabelled ionic exchange resin, or the small dosing volume used. 99mTc-labelled cholestyramine resin was administered in two dosage forms, a freeze-dried tablet which dissolved in the oral cavity (orally dissolving tablet; ODT) and a 1.5 mL aqueous suspension. Two resin particle sizes (20-40 and 90-125 microns) were studied. Oesophageal transit and intragastric distribution and residence were followed by gamma scintigraphy. In a second study, in six subjects, gastric emptying of the water-soluble fraction of the ODT and 1.5 mL of water, was measured using 99mTc diethylenetriaminepentaacetic acid. Oesophageal transit of a water-soluble marker and resin in suspension was rapid, but the transit of the resin in the ODTs was significantly prolonged. Regardless of particle size or dosage form, the resin was evenly distributed throughout the stomach with 20-25% remaining for 5.5 h. In contrast, the water-soluble marker cleared from the stomach rapidly from both dosage forms. We suggest that oral dose forms containing finely-divided ion-exchange resins may form a useful system for topical treatment of the gastric mucosa, for example in targeting to Helicobacter pylori infection. PMID- 8708985 TI - Involvement of NK1 and NK2 receptors in pulmonary responses elicited by non adrenergic, non-cholinergic vagal stimulation in guinea-pigs. AB - Previous studies from our laboratory using exogenously administered neurokinin (NK) agonists have shown that both NK1- and NK2-receptor subtypes are involved in plasma extravasation in the guinea-pig airways. In the present study, we have extended these observations using antidromic vagal stimulation to stimulate sensory c-fibres as a means of eliciting the release of endogenous tachykinins in propranolol- and atropine-treated guinea-pigs. Antidromic vagal stimulation (5 ms, 30 s) induced frequency-dependent (1-10 Hz) bronchoconstriction that was completely abolished by co-administration of the NK1-selective antagonist CP 99,994 ((2s-methoxy-benzyl)-(2-phenyl-piperidin-3s-yl)-amine), and the NK2 selective antagonist SR-48,968 ((S)-N-methyl-N-[4-(4-acetylamino-4-phenyl piperidino)-2-(3,4-dichlorophenyl) butyl]benzamide), each at a dose sufficient to block NK1 and NK2 receptors, respectively (each at 0.3 mg kg-1, i.v.). In contrast, SR-48,968 when given alone only partially blocked the vagal stimulation induced bronchospasm, whereas CP-99,994 had no effect. Significant increases (2-3 fold) in plasma extravasation of [125I]fibrinogen in the trachea, main bronchi, distal airways and oesophagus following vagal stimulation (5 Hz, 5 min, 10 V, 5 ms) were observed. Pretreatment with the neutral endopeptidase inhibitor, thiorphan (1 mg kg-1, i.v.), and the angiotensin-converting enzyme inhibitor, enalapril (1 mg kg-1, i.v.), potentiated both vagal stimulation-induced bronchoconstriction and plasma leakage in all tissues examined. This potentiation was due to reduced metabolism of endogenously released tachykinins since enhanced plasma overflow of immunoreactive substance P was observed following vagal stimulation in thiorphan- and enalapril-treated guinea-pigs. CP-99,994 substantially blocked plasma leakage in all parts of the airways and in the oesophagus. In comparison, SR-48,968 had no significant effect in the trachea and the oesophagus but partially inhibited plasma leakage in the main bronchi and distal airways. Co-administration of both CP-99,994 and SR-48,968 abolished the residual plasma leakage in these two regions. These results support the hypothesis that both NK1 and NK2 receptors are involved in tachykinin-induced pulmonary responses in the airways. PMID- 8708987 TI - Pre- and postjunctional effects of diadenosine polyphosphates in the guinea-pig vas deferens. AB - The pre- and postjunctional activities of a number of diadenosine polyphosphates were examined in the guinea-pig isolated vas deferens at the level of the membrane potential, using a modified sucrose-gap technique. P1,P3-Di(adenosine 5')triphosphate (Ap3A), P1,P4-di(adenosine 5')tetraphosphate (Ap4A) and P1,P5 di(adenosine 5')pentaphosphate (Ap5A) all caused concentration-dependent depolarization of the smooth muscle membrane. The potency order was: Ap5A > Ap4A > or = Ap3A. P1,P2-Di(adenosine 5')pyrophosphate (Ap2A) did not evoke depolarization even at the highest concentration tested (1 mM). All the dinucleotides caused a reduction in the amplitude of evoked excitatory junction potentials (e.j.ps). The potency order was: Ap5A = Ap4A > Ap3A > Ap2A. The depolarizations evoked by the dinucleotides were markedly reduced by the selective P2X-purinoceptor antagonist, pyridoxalphosphate-6-azophenyl-2',4' disulphonic acid (PPADS, 10 microM), as was the amplitude of the fully facilitated e.j.p. The inhibition of the e.j.p. evoked by Ap3A and Ap2A was reduced by the P1-purinoceptor antagonist, 8-p-sulphophenyltheophylline (8-pSPT, 50 microM), but that evoked by Ap5A and Ap4A was not. Thus, Ap3A, Ap4A and Ap5A evoke depolarization of the guinea-pig vas deferens via P2X-purinoceptors, and additionally Ap2A and Ap3A exert a prejunctional effect via P1-purinoceptors. The prejunctional activity of Ap4A and Ap5A is mediated via an undefined purinoceptor, which is neither P1 nor P2X. PMID- 8708986 TI - Effect of selective inhibition of potassium channels on vasorelaxing response to cromakalim, nitroglycerin and nitric oxide of canine coronary arteries. AB - A comparative study was performed on the sensitivity of in-vitro vasorelaxation by nitroglycerin and cromakalim to block glibenclamide, a blocker of ATP sensitive potassium channels, and iberiotoxin, a selective inhibitor of large conductance calcium-activated potassium channels. In isolated canine coronary arteries preconstricted with 25 microM prostaglandin F2 alpha, nitroglycerin (0.005-1.8 microM) and cromakalim (0.15-9.6 microM) produced dose-dependent vasodilations. Glibenclamide (30 microM) had no significant effect on relaxation of the dose-response curve to nitroglycerin and almost completely abolished the relaxation by cromakalim, a known opener of ATP-sensitive potassium channels. Iberiotoxin (90 nM) decreased the maximal response to nitroglycerin and had no effect on the vasodilation induced by cromakalim. The effect of iberiotoxin on the vasorelaxing action of nitric oxide, the active metabolite of nitroglycerin, was also examined. In a low potassium chloride (14.4-20.4 mM) medium, as a contractile stimulus, iberiotoxin inhibited relaxations by exogenous nitric oxide (100-200 nM). Enhancement of potassium concentrations to 35.4-40.4 mM significantly decreased relaxation by nitric oxide and under these conditions the inhibitory action of iberiotoxin disappeared. The present study demonstrated that in canine coronary arteries, the functional role of two potassium channels can be separated by pharmacological means. Nitroglycerin-induced vasorelaxation may be mediated, at least in part, by its enzymatic breakdown product, nitric oxide that activates large-conductance calcium-activated potassium channels. PMID- 8708988 TI - Prazosin-induced blockade of extraneuronal uptake facilitates dopaminergic modulation of muscle twitches in rat vas deferens. AB - Preliminary findings in our laboratory have shown that prazosin augmented the inhibitory effects of dopamine on the electrically-evoked muscle twitches in rat vas deferens. In this study, we opted to investigate the underlying mechanism and whether a prazosin-induced blockade of extraneuronal uptake process may be involved. Cumulative additions of dopamine (1.8 x 10(-7)-4.4 x 10(-5) M) elicited slight (< 30%) but dose-related inhibition of electrically-evoked (0.05 Hz, 1 ms duration and supramaximal voltage) muscle twitches of the vas deferens. Pretreatment with cocaine (10 microM), prazosin (50 nM) or oestradiol (10 microM) produced comparable potentiation of the inhibitory responses of dopamine; the pD2 values to dopamine amounted to 4.47 +/- 0.20, 4.72 +/- 0.21 and 4.56 +/- 0.19, respectively. A lower concentration of prazosin (5 nM) failed to alter dopaminergic responses. Further potentiation of dopamine responses was demonstrated in tissues preincubated with a combination of cocaine plus prazosin (50 nM), or cocaine plus oestradiol (pD2, 5.40 +/- 0.11 and 5.42 +/- 0.05, respectively). However, a mixture of all three drugs failed to elicit any further increase in dopamine responses, a finding that may suggest an extraneuronal uptake blocking activity for prazosin. Inhibition of muscle twitches evoked by bromocriptine, a dopaminoceptor agonist which is not a substrate for extraneuronal uptake, was not affected by prazosin (50 nM) pretreatment. The findings presented in this study emphasize the role of dopamine in modulating noradrenergic neurotransmission in rat vas deferens. More importantly, the results suggest that prazosin may act to block the extraneuronal uptake at noradrenergic sites, an effect that may account for its capability to facilitate dopaminergic modulation of noradrenergic neurotransmission. PMID- 8708989 TI - The effect of co-administered drugs on oxaprozin binding to human serum albumin. AB - The binding of the non-steroidal anti-inflammatory drug oxaprozin to human serum albumin was studied by bioaffinity high-performance liquid chromatography using a column based on immobilized human serum albumin. Displacement studies using marker compounds for the major drug binding sites showed that oxaprozin has a high affinity for the benzodiazepine/indole site and binds to the warfarin site but with a significantly lower affinity. Biochromatography and ultrafiltration techniques were used to screen for possible competition and allosteric interactions between oxaprozin and potential co-administered drugs including non steroidal anti-inflammatory drugs, antipyretics, hypoglycaemics, inhibitors of angiotensin-converting enzyme, anaesthetics, metal ions and anticancer agents. Competition occurred mainly with drugs bound at the benzodiazepine site (benzodiazepines, various non-steroidal anti-inflammatories). PMID- 8708990 TI - Bioavailability of phenytoin following oral administration of phenytoin-lipid conjugates to rats. AB - The bioavailability of phenytoin was evaluated in rats upon oral administration of phenytoin-lipid conjugates obtained by covalent binding of 3 hydroxymethylphenytoin to 1,3-dimyristoylglyceride via a succinidyl linkage, to 2 (1,3-dimyristoyl-2-glyceryl)butyric acid and to 3-myristoyloxy-2-methylpropionic acid. Despite differences of the phenytoin plasma concentrations all three compounds approximately doubled the AUC compared with the dosing of phenytoin itself. The early onset and the long duration of the anticonvulsant activity after administration of the triglyceride-derived conjugate could be correlated to the increased phenytoin plasma levels. It is concluded that drug-lipid conjugates may be useful prodrugs for the oral delivery of poorly water-soluble drugs. PMID- 8708991 TI - Availability and mean transit times of phenol and its metabolites in the isolated perfused rat liver: normal and retrograde studies using tracer concentrations of phenol. AB - Phenolic compounds are frequently detoxified by the formation of sulphate and glucuronic acid conjugates in the liver. These conjugates are formed in the hepatocytes and then either transported into the bile or back into the blood. In this study, we examined the transport kinetics of phenol and its metabolites in the isolated perfused rat liver by monitoring the outflow profiles of these compounds after a bolus input in a single pass preparation. Phenol was almost exclusively metabolized to phenyl sulphate (97%) at the trace concentrations used, with the amount of phenol and metabolites excreted into the bile being minimal (3.5%). The metabolite formed was rapidly transported back into the perfusate, with mean transit times of 17.4 and 12.3 s anterograde and 24.9 and 24.2 s retrograde at flow rates of 15 and 30 mL min-1 respectively, which were intermediate between those of Evans blue and water. The outflow concentration time profile for phenyl sulphate formation was unaffected by the addition of another organic anion (bromosulphophthalein). The effect of enzyme zonation on outflow concentration-time profiles was also investigated using retrograde perfusions. The transit time ratios for generated metabolite to water for anterograde perfusions (0.6) was found to be more than twice that for retrograde perfusions (0.23) at 15 mL min-1 and approximately 1.6 times greater at 30 mL min 1, being 0.58 and 0.37 respectively. The relative ratios obtained are consistent with previous findings that normalized variance of solutes in the retrograde perfusions is greater than that for anterograde perfusions. PMID- 8708992 TI - Metabolism of quinine in man: identification of a major metabolite, and effects of smoking and rifampicin pretreatment. AB - Our previous studies have shown that cigarette smoking and rifampicin pretreatment enhance the elimination of quinine, metabolism assumed, by analogy with quinidine, to be carried out by CYP3A (P450IIIA). This finding is unexpected since it has been shown that smoking induces the CYP1A rather than the CYP3A enzyme family, suggesting that the metabolism of quinine may be catalysed by CYP1A. Therefore, we conducted this study to identify possible quinine metabolites in human urine and to determine which metabolic pathway is induced by cigarette smoking and rifampicin pretreatment. A specific HPLC procedure was employed to identify metabolites of quinine in urine samples collected from healthy volunteers after an oral dose of 600 mg quinine sulphate. The results showed that there were at least seven possible metabolites of quinine detected in human urine. Three of these were identified as 2'-oxoquininone, quinine glucuronide and 3-hydroxyquinine. The 3-hydroxyquinine appeared to be a major metabolite of quinine in urine samples from every subject who took an oral dose of quinine. Although cigarette smoking and rifampicin pretreatment were shown to cause a marked increase in the elimination of quinine there were no significant changes in the formation of 3-hydroxyquinine as measured in the urine samples. This suggests that the effects of smoking and rifampicin are more complicated than we expected and require further investigation. PMID- 8708993 TI - Frusemide and its acyl glucuronide show a short and long phase in elimination kinetics and pharmacodynamic effect in man. AB - The pharmacokinetics of 80 mg frusemide given orally were investigated in normal subjects using a direct HPLC method for parent drug and its acyl glucuronide conjugate. Two half-lives could be distinguished in the plasma elimination of both frusemide and its conjugate, with values of 1.25 +/- 0.75 and 30.4 +/- 11.5 h for frusemide and 1.31 +/- 0.60 and 33.2 +/- 28.0 h for the conjugate. The renal excretion rate-time profile showed two phases; the rapid elimination phase lasted from 0-15 h and the second and slow phase, from 15-96 h. During the first 15 h, 33.3 +/- 4.8% of the dosed frusemide was excreted; in the remaining period 15-96 h, 4.6 +/- 1.5% was excreted. In the same two periods the excretion of the glucuronide was 13.4 +/- 4.7 and 1.9 +/- 1.1%, respectively. The mean renal clearance of frusemide was 90.2 +/- 16.9 mL min-1 during the first period and 91.5 +/- 29.3 mL min-1 in the remaining period, during which the stimulation of urine production was absent. The renal clearance of the acyl glucuronide was 702 +/- 221 mL min-1 in the first period, but only 109 +/- 51.0 mL min-1 in the second period. The stimulated urine production in the first 6 h after administration amounted to 2260 +/- 755 mL (measured urine production minus baseline value of 1 mL min-1 (360 mL). During the second or rebound period (6-96 h after drug administration), the quantity of urine was 990 +/- 294 mL lower than what would have been expected from the baseline production of 5400 mL. This reduced production (0.82 mL min-1) is equivalent to an 18% reduction in the average urine flow rate of 1 mL min-1. PMID- 8708994 TI - Preliminary efficacy study of a bioadhesive vaginal metronidazole tablet in the treatment of bacterial vaginosis. PMID- 8708995 TI - Meanings of political participation among black and white women: political identity and social responsibility. AB - This study examined the correlates of midlife political participation among 64 Black and 107 White women of the college classes of 1967-1973. Compared with White women, Black women scored higher on political participation, generativity, power discontent, and politicization. Factor analysis of personality and political attitude variables yielded three factors labeled Political Identity, Power Discontent, and Social Responsibility. Adult political participation was regressed on level of student activism and index scores of political identity, power discontent, and social responsibility. For both racial groups, social responsibility was associated with midlife political participation. For White women, political identity was also related; for Black women, student activism bore a significant relationship. The findings suggest that Black and White women's historical and political contexts imbued their political activities with different meanings. PMID- 8708996 TI - Pregnancy decision making as a significant life event: a commitment approach. AB - A value-laden basis for commitment to goals and the behavioral and affective sequelae of commitment were examined in the context of a stressful life event. Fifty-seven women who were interviewed during a clinic visit for a pregnancy test (Time 1 [T1]) subsequently received positive test results and were then interviewed 2 days later (Time 2 [T2]) and a month later (Time 3 [T3]). The intentionality and the meaning of the pregnancy were associated with self reported commitment to the pregnancy at T1. In turn, commitment predicted affective states both prior to (T1) and shortly after (T2) receiving test results. Initial commitment also predicted decisions to continue versus to terminate the pregnancy. The decision to continue the pregnancy appeared to bolster self-reported commitment. Relatedly, those continuing the pregnancy reported smoking fewer cigarettes at T3 than at T1. For those aborting the pregnancy, commitment at T1 was negatively related to adjustment at T3. Initial commitment predicted subsequent depression, guilt, hostility among those who aborted, whereas commitment predicted anxiety among those who continued the pregnancy. Other correlates of commitment (pregnancy concerns, religion, abortion history, and other life goals) were explored. PMID- 8708997 TI - Personal ideology polarity: its emotional foundation and its manifestation in individual value systems, religiosity, political orientation, and assumptions concerning human nature. AB - Personal ideology is an individual's philosophy of how life should be and of what forces influence human living. In this study S.S. Tomkins's (1963b, 1965, 1978, 1987) polarity theory of ideology was used to examine the manifestation of personal ideology in 4 value-laden domains of personality. Tomkins's theoretical postulates concerning the emotional foundation of personal ideology also were tested. Analyses revealed that the 2 defining dimensions of polarity theory- humanism and normativism--are related in the predicted meaningful ways to value systems, assumptions concerning human nature, religiosity, and political orientation. Evidence also was found for Tomkins's contention that specific affect clusters serve as the foundation of personal ideology. Participants who scored high in humanism ideology related autobiographical memories containing the emotion cluster of joy, distress, fear, and shame, whereas the memories of participants with normative ideologies contained relatively more anger. PMID- 8708998 TI - Hard times and hurtful partners: how financial strain affects depression and relationship satisfaction of unemployed persons and their spouses. AB - The process linking unemployment and economic hardship to depression and marital or relationship satisfaction in couples was examined. Using structural equation modeling, the authors tested models in which financial strain and partners' symptoms of depression influence the behavioral exchange of the couples in terms of social support and social undermining and, in turn, the effects of support and undermining on relationship satisfaction and depressive symptoms. The analyses were based on longitudinal data from 815 recently unemployed job seekers and their spouses or partners. The results demonstrated that financial strain had significant effects on depressive symptoms of both partners, which in turn led the partner to withdraw social support and increase social undermining. Reduced supportive and increase undermining behaviors had additional adverse effects on satisfaction with the relationship and on depressive symptoms. PMID- 8708999 TI - Depression and the explanation of events that happen to self, close others, and strangers. AB - Depressive and nondepressive college students attributed causality for positive and negative events that happened to either themselves, a close other, or a typical student. Depressives made less optimistic attributions than nondepressives when explaining events that happened to themselves. However, depressives and nondepressives generally made similar attributions about others; both groups were optimistic when explaining events that happened to their best friend or romantic partner and less optimistic when explaining events that happened to the typical student. The results indicate that depressives do not treat close others as extensions of the self, at least in terms of their attributional patterns. Furthermore, depressives were aware of the extent to which their attributions benefitted or harmed the desired identity of the actor. PMID- 8709000 TI - Status, communality, and agency: implications for stereotypes of gender and other groups. AB - Four studies addressed the hypothesis, based on correspondence bias, that low- relative to high-status individuals are perceived as more communal and less agentic. Study 1 instantiated status in terms of occupations, and findings were as expected. The findings of Study 2 reconciled those of Study 1 and of A.H. Eagly and V.J. Steffen (1984) in that they demonstrated that high-status occupations are differentially construed in terms of their interpersonal communal demands. The hypothesis received clear support in Studies 3 and 4, in which a general instantiation of status independent of occupations, social roles, and gender was adopted. The findings are discussed in terms of gender stereotypes and social role theory of gender (A.H. Eagly, 1987) as well as in terms of other stereotypes. PMID- 8709001 TI - Effects of mood during exposure to target information on subsequently reported judgments: an on-line model of misattribution and correction. AB - Three experiments investigated the effects of participants' mood during exposure to target information on delayed judgments of the target. Participants were exposed to a mood induction immediately before they acquired information about a political candidate and then reported their evaluation of the candidate at a later time. Effects of mood on judgment were moderated by 2 individual differences measures that can be interpreted in terms of processing efficiency. These were political expertise and total recall for the candidate information, with higher scores on these indices interpreted as reflecting more efficient processing. Among low-expertise (or low-recall) perceivers, mood produced an assimilation effect on evaluative judgments. Among high-expertise (or high recall) perceivers, mood produced a contrast effect on judgments. When pooling across these individual differences, mood exerted no influence on judgments. These findings are consistent with an on-line model of mood misattribution and overcorrection. PMID- 8709002 TI - Gender patterns in African American and white adolescents' social interactions in same-race, mixed-gender groups. AB - Gender-related interaction patterns on a cooperative problem-solving task were compared in African American and White mixed-gender groups of adolescents, matched on age, socioeconomic status, and school achievement. Hierarchically nested analyses of variance revealed significant interaction effects indicating greater gender equality in the African American groups than in the White groups for a composite measure of all variables, for the measure of level of activity, and for 1 of 2 measures of level of influence, but not for measures of speech form. Post hoc analyses revealed significant gender differences in the White groups for the composite measure, the level of influence measures, 1 speech-form measure, and the level of activity measure (nearly significant) and in the African American groups only for 2 speech-form measures. Results are discussed in terms of social role theory. PMID- 8709003 TI - [Establishment of fundamentals for protein research. Stabilization of protein]. AB - Proteins are usually in an equilibrium between the folded and the unfolded state. Therefore, for the stabilization of proteins against reversible denaturation, the free energy change for the unfolding should be increased by stabilizing the folded state by lowering the energy level of the folded state or by destabilizing the unfolded state by raising the energy level of the unfolded state. On the other hand, various processes can be coupled with the unfolded state of proteins. For example, protease digestion of proteins at physiological temperature may be one of such processes. The process would lead to an irreversible denaturation. For the stabilization of proteins against the irreversible denaturation coupled with the unfolded state, a kinetic stabilization is important, that is, the activation free energy for the unfolding should be increased. Methods for the kinetic stabilization were discussed. Finally, the irreversible chemical deterioration of proteins was considered. PMID- 8709005 TI - [Biochemical studies on functional proteins in the brain nervous system. Neurobiochemical studies on nerve growth factor]. AB - Nerve growth factor (NGF) is a polypeptide that is necessary for the survival and growth of developing sympathetic and sensory neurons as well as basal forebrain cholinergic neurons in the brain. The effects of NGF are mediated by the binding of the factor to its specific receptor present on the surface of NGF-responsive cells. Since NGF-responsive basal forebrain cholinergic neurons are lost in Alzheimer's disease, treatment with NGF may be therapeutically beneficial for the patients with this disease. Our studies were focused on the purification, amino acid sequence, enzyme immunoassay (EIA), NGF-biosynthesizing cells, regulation of the biosynthesis, compounds stimulating NGF-biosynthesis/secretion, etc. The major results obtained are summarized as follows: 1) We purified NGFs from snake venoms and determined their amino acid sequences. Among them, Crotalus adamanteus and Vipera russelli-NGFs were glycoproteins. 2) We developed highly sensitive (0.03-0.05 pg/tube or bead), simple, and reliable EIA systems for NGFs and NT-3. 3) Catecholamine, its derivatives, p-quinone derivatives, nicotine, nicotine derivatives, coenzymes, etc. enhanced the NGF-biosynthesis/secretion of fibroblasts and astrocytes by increasing their cellular content of NGF mRNA. 4) Basal forebrain-lesioned rats (an animal model of amnesia) administered 6-(4 hydroxybutyl)-2,3,5-trimethyl-1,4-benzoquinone (TMQ) orally showed improvement not only their NGF level and choline acetyltransferase (CAT) activity in the brain but also their memory and learning. 5) Interleukins (IL)-4 and -5 significantly increased the amounts of NGF secreted by mouse astrocytes; whereas IL-2, -3, and -6 had no significant effect. 6) Interferons-beta and -lambda inhibited the DNA- and NGF-synthesis in growing astrocytes cultured from neonatal mouse brain but they had no effect on the NGF-synthesis/secretion in quiescent astrocytes. 7) Alkylcatechols regulated the NGF gene expression in astrocytes via both protein kinase C- and cAMP-independent mechanisms. PMID- 8709004 TI - [Fundamentals of plasma chemistry and its application to drug engineering]. AB - In this review, our novel research works in both low temperature plasma chemistry and solid state plasma chemistry were described. As for low temperature plasma, the ESR study on plasma-induced radicals of several selected conventional polymers was shown including the detailed analyses of the radical structure and the mechanism by which the radicals were formed on typical degradable methacrylic polymers and cross-linkable polystyrene. One of the pharmaceutical applications of the plasma processing for drug delivery system (DDS) was also described, which includes the preparations of double-compressed tablet consisting of drugs as a core material and various types of polymers as a wall material followed by plasma irradiation on such a tablet. As for solid state plasma, the detailed reaction mechanism of solid state mechanochemical polymerization was shown including the solid state single electron transfer and the special feature of the resulting polymers. The structural criteria for polymerizable monomer derived from the quantum chemical considerations were also established. Based on the above findings, we synthesized various polymeric prodrugs by mechanochemical polymerization and studied the nature of hydrolyses (drug release). PMID- 8709006 TI - [Receptor-mediated signal transduction]. AB - The elucidation for the mechanism of receptor-mediated signal transduction has been the aim of our extensive studies. Cyclic AMP, which was controlled by membrane adenylyl cyclase, was an intracellular signal (the first second messenger in cells proposed by Sutherland) given by hormones and neurotransmitters. The GTP-binding (G) proteins play an important role in communication between membrane receptors and the adenylyl cyclase catalytic unit. One (Gs) of the G proteins is involved in the activation, while the other (Gi) is involved in the inhibition of adenylyl cyclase. Islet-activating protein (IAP, pertussis toxin, PTX) catalyses the transfer of the ADP-ribose moiety of NAD to the alpha subunit of Gi, resulting in a complete loss of the Gi functions. In some cases, arachidonic acid (AA) regulates cell functions as a second messenger. AA serves as a precursor to a number of biologically active lipids including prostaglandins and leukotrienes. Activation of cell surface receptors of many cell types results in the release of AA from membrane phospholipids by phospholipase A2 (PLA2). A new family of PLA2 has been discovered in the cytosol of various cells. The activation of receptor-mediated AA release by cytosolic PLA2 was also regulated by PTX-sensitive G proteins. PTX treatment inhibited cell growth of fibroblasts by serum and growth factors. G proteins have been involved in receptor-receptor interactions in neuronal cells. These findings suggest the regulatory roles of cell surface receptors-coupled G proteins in signal transductions and cell functions. PMID- 8709007 TI - [Augmented ability of spleen cells to produce interferons and prevention from lethal infection of herpes simplex virus in mice orally treated with Enterococcus faecalis preparation, FK-23]. AB - Effects of the oral or intraperitoneal administration of an Enterococcus preparation, FK-23, to mice on the interferon (IFN) production by their spleen cells and on the host defense against the infection with herpes simplex virus (HSV)-1 were examined. Spleen cells were obtained from the mice intraperitoneally treated with cyclophosphamide (CY) and subsequently orally administered FK-23 preparation, and then cultured with phytohemagglutinin-P or bacterial lipopolysaccharide in vitro. They produced higher titers IFN than those obtained from control mice which were not treated with the FK-23 preparation. The IFN activity was neutralized mainly by antiIFN-beta antibody. Correspondingly, oral (5 mg/mouse) or intraperitoneal (1 mg/mouse) administration of the FK-23 preparation protected some of the CY-pretreated mice from death by HSV-1 infection. PMID- 8709008 TI - [Quantitative analysis of chlorpheniramine maleate in cough and cold drugs by ion pair high-performance liquid chromatography for the simultaneous determination of chlorpheniramine and maleate]. AB - A simple, rapid and convenient chromatographic method, which permits the simultaneous determination of chlorpheniramine (CP) and maleate (MA), recently developed by the authors was applied to the analysis of chlorpheniramine maleate (CPM) in cough and cold drugs. In this method, a Capcell Pak C8 column and an isocratic mobile phase containing 15% methanol, 50 mM KH2PO4 and 5mM tetra-n butylammonium phosphate as an ion-pair reagent were used. By using the mobile phase adjusted to pH 3.0 with orthophosphoric acid, fumaric acid, MA, CP, acetaminophen (paracetamol), caffeine, and m- and p-hydroxybenzoic acid as candidates for an internal standard were eluted separately within 17 min. Detection was carried out with UV detector at 215 nm. Under the same conditions, five other antihistamines analogous to CPM were also separated. The calibration graphs for CP and MA showed good linearity in the range of 0.5-10 nmol (0.195-3.9 micrograms) per 20 microliters injection, respectively. The proposed method was successfully applied to the simultaneous determination of CP and MA, i.e., CPM analysis, in commercial cough and cold drugs which pharmaceutical forms were tablet, granule and syrups. PMID- 8709009 TI - Application of lingual contact occlusion to complete dentures with two-layered posterior teeth: a clinical report. AB - When two-layered artificial teeth are used, acrylic resin occlusal parts can be removed from base parts and used directly as resin patterns for casting, which facilitates fabrication of metal occlusal surfaces. Because these teeth were made of acrylic resin, it was easy to modify occlusal morphology before converting to metal. Application of two-layered artificial teeth to achieve lingual contact occlusion brought about sufficient masticatory forces and denture stability and resulted in patient satisfaction. PMID- 8709010 TI - Assessment in the treatment of hypernasal speech with motor neuron disease: a clinical report. AB - The application of nasal anemometry in this patient's treatment proved to be of value in obtaining an objective assessment of this intervention. Nasal anemometry may be of use in other situations, for example, (1) in preassessment and postassessment of cleft palate surgery and surgical interventions to improve the velopharyngeal seal (such as pharyngoplasty), (2) in the evaluation of nasalized speech with conventional speech therapy, (3) in the evaluation of nasalized speech by use of a palatal training appliance, and (4) in the investigation of snoring. PMID- 8709011 TI - Craniofacial osseointegration of a large midfacial bone-anchored combination maxillofacial prosthesis: a clinical report. AB - The combination of retentive clips and magnets has been used with favorable results for smaller facial prostheses. This patient's large midfacial defect created the need for an attachment design that would retain an intraoral obturator prosthesis and a large silicone facial prosthesis. The advantages of this design include having an obturator that is separately attached by clips to the framework. This allows the patient more ease in insertion and removal of the prosthesis. It also allows for better oral hygiene. The magnets aid the patient in the proper alignment of the facial prosthesis. The O-ring attachment was not a complicated technique in this patient's treatment. The patient can, without the use of a mirror, snap the rubber matrix O-ring attachment over the patrix attachment on the framework. This resulted in a retentive facial prosthesis, which helped the patient feel more secure. The silicone margins were well adapted to the patient's skin, which allows the use of thin translucent margins and the elimination of the inherent problems associated with the use of adhesives. PMID- 8709012 TI - Stress-induced cervical lesions: review of advances in the past 10 years. AB - The accumulation of experimental and clinical evidence during the past decade has significantly contributed to the understanding of the role of occlusally generated tensile stress in the etiology of certain noncarious cervical lesions of teeth. More important, this knowledge has led to the understanding of the reasons why traditional restorative treatments of these stress-induced cervical lesions fail. The case of failure can be attributed to the occlusally generated stresses that are concentrated at the cervical region and result in debonding, leakage, retention failure, and, ultimately, restorative failure. With the new understanding, restorative approaches that combine chemical adhesion and restorative materials of appropriate elastic properties show promise of long-term success. PMID- 8709013 TI - Microleakage of a 4-methacryloxyethyl trimellitate anhydride bonding agent with amalgams. AB - Microleakage contributes to deterioration of materials, recurrent decay, growth of microorganisms at the interface, and postoperative tooth sensitivity. This study examined the microleakage between amalgams and tooth surfaces with a cavity liner, a 4-methacryloxyethyl trimellitate anhydride bonding agent (Amalgambond) and six different types of amalgams. Class 1 cavity preparations in extracted human molars were filled with different combinations of liners and amalgams and were stored at 37 degrees C in physiologic saline solution. After 1 week half of each amalgam restoration was removed. Within the same group the same cavity treatment was performed and the same type of amalgam was packed as before. Microleakage was determined after 2000 thermal cycles. Statistical analysis indicated that Amalgambond significantly reduced microleakage of different amalgams compared with the Copalite-lined and unlined controls. No microleakage was detected at the interface between the existing and replacement amalgams. PMID- 8709014 TI - Apical root strain as a function of post extension into a composite resin core. AB - This study tested the hypothesis that there is no difference in the amount of strain measured in the root of an extracted tooth at the apical end of a prefabricated post, whether a composite resin core completely covers the head of the post or the post is exposed at the occlusal surface of the core. Twenty extracted teeth were measured (1) after endodontic treatment (baseline), (2) after a composite resin core was fabricated over a prefabricated post, and (3) after a cast crown was cemented. The teeth were randomly divided into two groups after baseline testing. In group 1, the post was buried within the core, and in group 2, the head of the post was exposed on the occlusal surface of the core. Analysis of results indicated a statistically significant decrease in strain from the baseline to the crown when 1 mm of composite resin covered the head of the post. However, based on the small strain values measured, the difference may not be clinically significant. PMID- 8709015 TI - Retention and failure mode after cyclic loading in two post and core systems. AB - Retention of a prefabricated post and composite resin core system was compared with that of a cast post and core restoration in anterior maxillary teeth on which root canal preparations, enlarged in the coronal part, were performed. The effect of cyclic loading on restoration retention was also investigated. Retention of restorations not subjected to cyclic loading was not influenced by post type, but loaded prefabricated post restorations showed lower retention than both loaded cast post and core restorations and nonloaded prefabricated post restorations. After dislodgement, the failure mode was also determined. The results of this study suggest that if a canal requires extensive preparation, a well-adapted cast post and core restoration may be more retentive than a prefabricated post restoration that does not match the canal shape. PMID- 8709016 TI - A review of residual ridge resorption and bone density. AB - Residual ridge resorption is a common and often incapacitating problem, particularly for persons with edentulous mandibles. Several studies suggest a correlation between ridge resorption and osteoporosis. Recent radiologic studies indicate that the mineral density of the cortex and the bone mass in the mandible are correlated with skeletal bone density. Most resorption occurs in the alveolar process, whereas the basal portion remains relatively intact. On the other hand, radiologic measurements of bone density primarily yield information after the basal portion, where the bone mass of the mandible is greatest and functional stresses of mastication may affect bone density. Ironically, radiologic measurements may not accurately indicate the effect of osteoporosis on alveolar resorption. Apparently, excessive occlusal force can also produce extensive atrophy without systemic impact. Not until muscular function decreases does real osteoporosis develop in edentulous jaws. Not only does the volume of the ridge decrease, but also the density of the basal portion decreases as a result of diminished function. This article reviews the literature on residual ridge resorption and components that may affect the rate of resorption. PMID- 8709017 TI - Corrosion considerations in the brazing repair of cobalt-based partial dentures. AB - Cobalt-based alloys (Co-Cr-Mo) are usually used in dentistry as frameworks for removable partial dentures. In their basic form these structures function successfully. However, modifications or repairs of the frameworks may reduce their resistance to corrosion and, as a consequence, may provoke biologic reactions in the soft tissues. These reactions may be the result of different types of alloys that contact each other and, in the presence of saliva (based on potential differences), produce a galvanic cell. In this study, a clinical situation after repair of a removable partial denture was examined. The metallographic study of the prosthesis revealed a brazed zone where a gold braze was joining the Co-Cr-Mo framework with a Co-Cr-Ni type alloy (without Mo). The latter revealed signs of corrosion. Various electrochemical parameters (Ecorr, Ecouple, icorr, icouple) of these alloys were analyzed in the laboratory. The Co Cr-Ni alloy had the lowest nobility and underwent galvanic corrosion in a galvanic couple with gold braze. PMID- 8709018 TI - Designing a transmandibular implant with the aid of biostereometric analysis. AB - A biostereometric survey of severely resorbed dry mandibles was used to determine whether it was possible to design a transmandibular implant that could be used for the majority of clinical situations. Computerized profiles of the relevant mandibular areas were generated and superimposed to examine the feasibility of a universal shape. It would appear that the crestal shape of the mandible between the mental foramina is on the circumference of a circle and that the surface of the inferior border is so variable that a flat surface would have to be used for a transmandibular implant. A design of a transmandibular implant is proposed on the basis of the common anatomic features of the mandible. PMID- 8709019 TI - Magnetic resonance imaging findings in asymptomatic volunteers and symptomatic patients with temporomandibular disorders. AB - Internal derangement has been suggested to be a cause of temporomandibular disorders. This study compared magnetic resonance imaging findings in 82 asymptomatic volunteers and 263 symptomatic patients. Disk displacement was observed in 33% of the asymptomatic volunteers and 84% of the symptomatic patients. The prevalence of trauma, headache, and other joint problems was more common in symptomatic patients. A history of general anesthesia and orthodontic treatment was not increased in the symptomatic population compared with asymptomatic volunteers. This study suggests that disk displacement is more common in symptomatic subjects. PMID- 8709020 TI - Temporomandibular joint dysfunction and flat lateral guidances: a clinical association. AB - Temporomandibular joint alterations have been associated with abnormal mandibular movements in the open/close cycles and in the laterotrusive border movements. The quantitative analysis of these movements could allow a better or earlier diagnosis for patients and offer some insight into the pathophysiology of the disorder. Maximum opening and mandibular laterotrusive border movements were studied and measured in a group of 165 patients with a considerable lateral deviation (equal to or greater than 5mm) in maximum opening. Movements were directly performed by the patients and recorded with a mandibular kinesiography. The slope of the lateral guidance (frontal plane projection) was measured in the first millimeters of motion and started from the maximum intercuspal position. On average, slopes were significantly flatter on the same side with the opening deviation. These slopes were significantly flatter than previously published physiologic ranges, whereas contralateral slopes were similar to normal references. The results may be explained by a correlation between the insufficient lateral protection and a temporomandibular mandibular joint dysfunction (ipsilateral low mobility). PMID- 8709021 TI - Marginal discrepancy of ceramic crowns with redesigned implant components. AB - A previous study indicated a mean marginal discrepancy of 169 microns (SD = 23) for cemented CeraOne ceramic crowns. Since then, the components have been redesigned by the manufacturer. This study assessed the marginal discrepancy of the new components by use of a video camera and stereomicroscope connected to a computer. Fifteen samples each of (a) ceramic caps (control), (b) ceramic caps fired once in a porcelain oven (c) ceramic crowns, and (d) cemented (zinc phosphate) ceramic crowns were examined. The results indicated that the mean marginal discrepancies were: caps 20.4 microns (SD = 10.8), fired caps 18.3 microns (SD = 9.3), crowns 28.1 microns (SD = 9.7), and cemented crowns 65.9 microns (SD = 15.8). Statistical analysis (ANOVA) revealed significant differences between these mean marginal discrepancies (p < 0.0001). A Scheffe F test revealed no statistically significant difference between the means for caps, fired caps, and crowns, whereas cemented crowns had significantly greater mean marginal discrepancies compared with all uncemented retainers (p < 0.0001). The mean marginal discrepancy of the redesigned CeraOne ceramic system (cemented crowns) was reduced by more than 100 microns compared with the original design. PMID- 8709022 TI - The effect of posture change on the position of the skin marks for the transverse horizontal axis. AB - Marks are placed on facial skin in clinical dentistry to indicate the position of more deeply placed landmarks or reference points. In this study the movement of the transverse horizontal axis skin points overlying the craniomandibular articulation were observed and quantified. The time taken for the skin displacement to occur when the posture changed between upright and supine was also studied. The extent of displacement of the skin point was approximately 3 mm in the sagittal plane and 2 mm in the frontal plane when the posture was changed from upright to supine and vice versa. The displacement was complete after 30 seconds in 95% of subjects. The direction of the displacement was primarily cephalad but with a dorsal component of more than 10 degrees in 87% of subjects. The extent of the movement in the sagittal and frontal planes was correlated. There was no gender difference for the skin displacement. Awareness by clinicians of the extent and direction of such facial skin movements can help to prevent errors. PMID- 8709023 TI - Cellular inflammatory responses to implanted dental materials. AB - Cellular inflammatory responses to subcutaneous implantation of amalgam and composite resins were assessed in rats by use of histologic and immunocytochemical methods 2 days to 8 weeks after implantation. Frozen and paraffin sections were obtained from subcutaneous tissues associated with amalgam and composite resin suspensions. The amalgam induced mild inflammation with proliferation of few resident macrophages, but implantation of composite resins was associated with an influx of monocytes, increased numbers of resident connective tissue macrophages, and abnormal major histocompatibility antigen class II (Ia antigen) expression. The data suggest that composite resins may produce a a more pronounced inflammatory response than dental amalgams do when incorporated in soft tissues. PMID- 8709024 TI - Surgical guide fabrication for an angled implant. AB - This article describes the steps necessary for determining ideal implant abutment position and, subsequently, the optimum implant location that would make this abutment position possible. Procedures necessary for fabricating a surgical guide are explained for placing an ITI angled implant. PMID- 8709025 TI - Preventing loosening of implant abutment screws. AB - A procedure for using dimples inside the abutment screw cylinder above the screws and filling the holes with elastomeric impression material will prevent the screw retained prosthesis from loosening. A modification of the procedure can also be used for angled abutments. PMID- 8709026 TI - The effect of simulated function on the retention of bar-clip retained removable prostheses. AB - Patients wearing bar-clip retained removable prostheses may have loss of retention because of changes within the bar-clip assembly. This in vitro study recorded and compared the retention of one- and two-clip retained simulated mandibular complete denture prostheses before and after simulated function. Cast metal Hader bars and clip holders were used to make 10 one-clip and 10 two-clip specimen pairs. Tensile removal values before and after simulated function were recorded and compared by repeated-measures analysis of variance and Student tau tests (significance level 0.05). The results revealed that the use of two clips instead of one significantly increased retention of the simulated prosthesis. It was also found that there was a significant loss of retention after the specimens were placed on the bars and then removed once for both the one- and two-clip groups. Simulated function did not cause a significant change in retention for either group. PMID- 8709027 TI - Effects of chlorine-containing disinfecting compounds on shade guides made of acrylic resin. PMID- 8709028 TI - Fabricating a spray gun for opaque porcelain application. PMID- 8709029 TI - The modification of implant tightening components for use with torque drivers. PMID- 8709030 TI - A modified incisal guide pin (see-pin). PMID- 8709031 TI - Verifiable method for registering the centric relation position in dentulous arches with a central bearing point. PMID- 8709032 TI - Conscious sedation: a multidisciplinary team approach. AB - The administration of intravenous conscious sedation for patients undergoing minor procedures has increased in many settings throughout health care. Recognizing this, managers are challenged to develop policies to standardize the quality of patient care delivery. Consumer interest and knowledge in health care has increased dramatically in the last few years. Their demands for decreased cost, increased technology, and quality of care have compelled health care organizations to adopt quality management models. Quality improvement teams were developed to look at processes crossing departmental boundaries. Teams facilitate dialogue, understanding, and knowledge, and use the scientific method to design, streamline, and improve processes. The authors share strategies for policy development using a 7-step quality improvement process with a multidisciplinary team approach. PMID- 8709033 TI - Duplicate publication: guidelines for nurse authors and editors. PMID- 8709034 TI - Incidence of diuresis following hysterectomy. AB - Despite literature documenting ureteral obstruction associated with certain gynecological conditions, the occurrence of diuresis among posthysterectomy patients has not been examined. This nonexperimental exploratory study investigated whether diuresis was greater following hysterectomy for conditions that could potentially cause obstruction of the urinary tract than for other conditions requiring hysterectomy. Roy's adaptation model was used to emphasize the importance of expedient detection and intervention of the resultant fluid volume deficit. A retrospective chart review of 140 posthysterectomy patients was conducted. Four strata were used to divide the sample records into one nonobstructive and three potentially obstructive conditions for surgery. A ratio of total intake divided by total output was formulated for each patient and compared between the groups. Statistically significant differences were shown by one-way analysis of variance between two of the obstructive groups and the nonobstructive group (F = 5.182; P < .05). Results of this research showed that certain obstructive conditions--uterine prolapse/endometrial cancer and leiomyomas greater than 15 week size--were linked with substantially higher postoperative urine outputs, which increase the risk of hypovolemia. PMID- 8709035 TI - Care of patients after aortic aneurysm repair. AB - The purpose of this article is to identify indications for aortic aneurysm repair, review pathophysiology, and identify postoperative nursing priorities for patients following aortic aneurysm surgery. PMID- 8709036 TI - Reading research critically: results: multiple regression analysis. AB - Multiple regression is an analysis tool used much more frequently than bivariate regression analysis in the research we are reading. This article is designed to help the reader understand multiple regression analysis and confidence intervals. PMID- 8709037 TI - Core culture: the key to successful paradigm shifts. AB - The core culture of an organization is a major force to be recognized when trying to implement any type of change. Those changes that do not fit well with an existing culture will be strongly resisted. The very culture that may have to undergo change to survive the many paradigm shifts occurring in health care were the basis for past successes. The ability to recognize the core culture of an organization or unit will provide the manager with valuable information when the plan calls for implementing change. PMID- 8709038 TI - Patient discharge from the ambulatory setting. AB - Ambulatory surgical care is an integral part of the health care spectrum. The advantages of such care are well documented. One benefit of outpatient care is the potential for cost savings for the patient, medical facility, and third party payers by dismissing the patient to a remote recovery location following surgery and intermediate anesthetic recovery. To realize this goal an essential component of patient management must be the safe and expedient postoperative care of the patient until they are discharged from the ambulatory facility. This article will review considerations for discharging the surgical patient from the ambulatory setting. PMID- 8709039 TI - Rewarming hypothermic postanesthesia patients: a comparison between a water coil warming blanket and forced-air warming blanket. PMID- 8709040 TI - What's in a name? PMID- 8709041 TI - Latex allergy: recognition and perioperative management. AB - Because more than 40,000 household and medical products now contain latex, it is difficult if not impossible to avoid. Repeated exposure to latex leads to increased sensitization in susceptible people, resulting in significant, sometimes life-threatening problems. Health care workers need to be aware of latex allergy for their patients' protection, as well as their own. Proper identification of a potential or known latex allergy, awareness and avoidance of products containing latex, and appropriate perioperative management can avert the tragedy of anaphylactic shock and death. PMID- 8709042 TI - Conducting research: getting started. AB - This article presents issues to be considered when planning to conduct research, specifically ways to avoid critical errors in writing the research proposal. It concludes with an outline for writing research proposals. PMID- 8709043 TI - It's time to answer the wake-up call. AB - The dramatic change in the work world of every industry including health care is creating anxiety and stress among workers. According to many authors, this is merely the tip of the iceberg in relation to what will come. The way we must do our work is changing at breakneck speed. Resisting it will not stop it, fearing it will not help. Learning to respond quickly and recognizing that technology will only replace the routine and systematic processes can restore the energy necessary to meet the challenges ahead. PMID- 8709044 TI - Notes from the American Society of Anesthesiologists annual meeting. PMID- 8709045 TI - President's page: crystal reflection. PMID- 8709046 TI - The postanesthesia care unit: a high-risk environment for bloodborne and infectious respiratory pathogens. AB - The PACU is a high-risk environment for exposure to infectious diseases. A confluence of risk factors unique to the PACU increases the probability for exposure of personnel to both bloodborne and airborne pathogens. These risk factors include frequent coughing in the PACU, blood-contaminated saliva in the PACU, air mixing maximized in the PACU, high patient census and rapid patient turnover, inadequate patient histories, and the proximity of the postanesthesia nurse to the patient's face. Both the Occupational Safety and Health Administration and the Centers for Disease Control have issued recommended procedures for limiting occupational exposure of personnel to these hazards. PMID- 8709047 TI - Using a time-flow study to identify ambulatory surgical delays. AB - Identification of surgical delays in an ambulatory care setting is essential for providing consumer-sensitive nursing care. One method of identifying and documenting surgical delays is described. This method, a time-flow study, allowed a freestanding ambulatory surgical center to identify problem areas within its scope of operations, and to provide a factual basis for implementing solutions as part of a continuous quality improvement and cost analysis program. PMID- 8709048 TI - The ophthalmic ambulatory surgery patient. AB - Eye surgery is performed on patients of all ages, with most of these procedures performed on an outpatient basis. Common opthalmic terminology and anatomy are presented. In addition, the intraoperative procedures and postoperative care following surgery to correct strabismus, cataracts, glaucoma, and retinal detachment are described. PMID- 8709049 TI - Drugs affecting hemostasis. AB - Most patients admitted to the PACU are closely observed for postoperative bleeding. Although some bleeding is to be expected, some patients may have alterations in the hemostatic process leading to excess blood loss. Hemostasis can be intentionally altered by drugs given preoperatively and/or intraoperatively. The effects of these drugs may contribute to the postoperative blood loss. This article reviews the normal hemostatic process as a foundation for discussing the commonly used drugs affecting hemostasis. PMID- 8709050 TI - Pharmacokinetics in postanesthesia recovery: implications for nurses. AB - As the number of drugs used in the treatment of patients increases, nurses must be more knowledgeable about their activity and actions. This article reviews principles of pharmacokinetics, which is the study of how drugs attain their concentrations in the blood stream. The phases of absorption, distribution, biotransformation, and excretion are presented, and examples of common drugs are related to these phases. PMID- 8709051 TI - Advertising medicines to the public. PMID- 8709052 TI - Should drug information be an integral part of health care? AB - The fact that the effects of drugs are incompletely known makes special demands on drug information for the public. Much of this information is marred by poor content, choosing target groups by likely sales, and giving drug-centred rather than health-centred information. These problems arise from the imbalance between the relative quantities of commercial and independent drug information, the separation of drug information from health education, and strong commercial and professional interests. Suggested solutions include setting standards, integration, and using pharmacists to distribute drug information. Such actions may, however, create new problems. It might be necessary or desirable to regulate the quantity, forms and sources of commercial drug information aimed at consumers. Drug information should be regarded as an integral part of health care and treated accordingly. Furthermore, the widening range of substances used to influence health, shrinking prescription requirements, biased reporting in mass media, and national industrial priorities are changing the drug information scene and demand new solutions. PMID- 8709053 TI - Informing the patient. AB - 1. Patients value most the information provided to them by doctors. Doctors do not seem to be able to get messages across as satisfactorily to lower social groups as to higher social groups, since they give less time per consultation and fewer explanations to working-class patients; they are the ones that are least influenced by health advice [24] and would benefit most from being given more time and attention in consultations. 2. The community pharmacists have made great play on becoming a major information source for the public but have not yet succeeded in this role. 3. In today's society individuals will have to take greater responsibility for their own healthcare. Patients must therefore actively demand more information from their doctors and pharmacists in order to make decisions. 4. Doctors and pharmacists must recognise that patients are often unaware of what they need to know. Patients' intelligence must never be under estimated or their knowledge and understanding over-estimated. 5. The manufacturer has a major role in providing information by means of the patient information leaflet which must be in terms that can be understood and influence the patient's health behaviour. Written information is the only information to which a patient can repeatedly refer to reinforce messages given by doctors or pharmacists. Written information is however of most value to social groups A and B and is less appreciated by other groups. PMID- 8709054 TI - Statistical malpractice. AB - Statistical malpractice is an insidious, and indeed prestige-laden and grant rewarded, activity. Brilliantly clever, but fundamentally wrong-headed, number crunchers are encouraged to devise inappropriate applications of mathematical methods to health problems. This species of misdirected zealot has so far been immune from criticism. PMID- 8709056 TI - The permanent vegetative state. Review by a working group convened by the Royal College of Physicians and endorsed by the Conference of Medical Royal Colleges and their faculties of the United Kingdom. PMID- 8709055 TI - Screening for hyperlipidaemia in childhood. Recommendations of the British Hyperlipidaemia Association. AB - Children with familial hypercholesterolaemia are at high risk of developing coronary artery disease in early adulthood. The diagnosis should therefore be made in childhood. Population screening identifies a small number of children with major genetically determined disorders of lipid metabolism and a large number with polygenic hypercholesterolaemia of uncertain prognostic significance. Selective screening based on a family history of familial hypercholesterolaemia or premature coronary artery disease is an appropriate strategy for identifying most children with familial hypercholesterolaemia. A non-fasting total cholesterol measurement is a suitable screening test: if the concentration exceeds 5.5 mmol/l, a fasting measurement of total cholesterol, high-density lipoprotein cholesterol and triglyceride is required. The diagnosis in a child under 16 years should be based on finding a total cholesterol concentration greater than 6.7 mmol/l and a low-density lipoprotein cholesterol concentration above 4.0 mmol/l on at least two measurements taken more than one month apart. Children should not usually be screened before the age of two years, but the aim should be to diagnose heterozygous familial hypercholesterolaemia before the age of 10 years. Affected children should be referred for specialist care. PMID- 8709057 TI - The diagnosis of epilepsies. AB - Vague labels such as "epilepsy', "grand mal' or "seizures' are detrimental to patient care, and should be universally abandoned in favour of the syndromic diagnosis of epilepsies. PMID- 8709058 TI - Bone density in asthmatic patients taking inhaled corticosteroids: comparison of budesonide and beclomethasone dipropionate. AB - We assessed bone mineral density (BMD) in 20 asthmatics who had been taking inhaled budesonide (BUD) (median daily dose 800 micrograms) for over a year, 13 of whom had taken previous courses of systemic steroids. Their results were compared with those of 20 patients receiving inhaled high-dose beclomethasone dipropionate (BDP) (median daily dose 1,000 micrograms), all of whom had received previous courses of systemic corticosteroids, and with those of 17 mild asthmatics who had never taken either inhaled or systemic steroids. Mean (standard deviation) (SD)) BMD in the patients taking BUD was 139.5 (28.6) mg/ml. This was significantly lower (p < 0.05) than in the control patients who had never taken inhaled or systemic steroids (160.4 (27.4) mg/ml). Mean BMD in the patients taking BUD did not differ significantly from that observed in patients taking BDP (127.5 (22.6) mg/ml). Although the reduction in BMD in the asthma patients taking regular high-dose BUD could have been due to previous courses of corticosteroid, the magnitude of bone loss is similar to that seen in patients taking high-dose inhaled BDP and intermittent corticosteroids. PMID- 8709059 TI - When can elderly patients be excluded from discussing resuscitation? AB - Case notes of elderly medical patients were surveyed to determine when "do not resuscitate' (DNR) decisions could legitimately be made without consulting them. Patients were thought to be suitable for exclusion from decisions if morbidity scores indicated that they were unlikely to survive cardiopulmonary resuscitation (CPR) or if they were mentally incompetent. Thirty per cent of all patients were predicted not to survive CPR; another 28% were deemed incompetent. Of those with DNR decisions, 59% were predicted not to survive and a further 24% were incompetent. Discussing resuscitation would have been appropriate with 17% of those with DNR decisions. PMID- 8709060 TI - Risk factor control five years after coronary bypass grafting. AB - The prevalence of three major coronary risk factors, hyperlipidaemia, hypertension and cigarette smoking, and the change in lipid fractions were assessed five years after coronary bypass surgery and compared to pre-operation in 353 consecutive patients undergoing elective first time surgery at a single centre. Five years after surgery 309 patients were alive without further cardiac surgery, questionnaire follow-up was available in 291 (94%). Lipid profile measurements were made in 255 (83%) of these five-year survivors and 100 (34%) had a history of hypertension. Five years after surgery, 146 (50%) of the 291 patients thought they had high cholesterol, of whom 92 (63%) were following a diet, 36 (25%) were also taking lipid-lowering drugs and 18 (12%) were taking no measures; 141 (48%) of them did not think they had high cholesterol, and four (2%) did not know. There had been a favourable change in all lipid fractions compared with pre-operation, including total serum cholesterol, particularly in patients taking lipid-lowering drugs. However, total serum cholesterol was above 5.2 mmol/l in 203 (80%) patients and low-density lipoprotein (LDL) cholesterol was above 3.4 mmol/l in 180 (71%). Only 30% of patients taking lipid-lowering drugs had an LDL cholesterol of 3.4 mmol/l or less. Blood pressure was recorded in 257 (83%) of the 309 five-year survivors: 82 (28%) were taking antihypertensive medication; 32 (12%) and 87 (34%) patients had a diastolic blood pressure above 95 mmHg and 90 mmHg, respectively, and 49 (19%) had a systolic blood pressure above 160 mmHg. There were 22 (9%) regular cigarette smokers. Corroboration of non-smoking with exhaled carbon monoxide measurements confirmed an accurate history of non-smoking in nearly all patients. PMID- 8709061 TI - The patient focused approach: a better way to run a hospital? AB - It can take 2 hours of hospital staff time to obtain a routine chest X-ray, up to 47 clinical staff may be involved with a patient during a 5-day stay, and only a quarter of total costs may be for direct patient care, so some hospitals are experimenting with patient focused care by relocating services such as X-ray to the bedside, training ward staff in a wider range of skills, and managing care itself by using multidisciplinary protocols. Potential benefits can be measured in terms of reduced process times and faster turn-round, but duplication of, for example, high-tech pathology and radiology equipment is expensive, as is releasing staff for training. Proponents say that higher quality patient care will result without increased cost, and theoretical analyses suggest that advantages should outweigh disadvantages. The more established patient focused units in the UK are now over a year old; practical analyses of their quality and cost are under way. PMID- 8709062 TI - Ward based X-ray facilities can improve services. AB - As part of a wider experiment, a satellite X-ray facility, run by 2 part-time radiographers (1 whole time equivalent), was established to provide all plain radiographs on inpatients in a patient focused unit of 114 beds created from 4 medical wards of a 370-bed district general hospital providing acute services to a local population of about 200,000. Fewer staff were needed to provide an X-ray and report on the ward, the number of steps was reduced from 54 to 42 and the time taken from 104 to 62 minutes. Radiographers spent a smaller proportion of their time on professional duties but freed up substantial time for other members of staff. The reactions of all involved, staff and patients, were favourable. With present technology, the patient focused approach brings net benefits and possibly decreases costs, but the balance may swing back to centralisation when picture archiving and communication systems (PACS) become more widely available. PMID- 8709063 TI - Obstetric physicians: are they needed? The workload of a medical complications in pregnancy clinic. AB - Obstetric physicians are physicians whose training and particular expertise lie in the care of the medical complications of pregnancy. We present an audit of the work of our bi-weekly obstetric medicine clinic at Queen Charlotte's Hospital, London, over a 10-month period from July 1994 to April 1995 inclusive, and discuss the need for obstetric physicians, with reference to individual medical conditions which complicate pregnancy. PMID- 8709064 TI - Science: an unnatural practice. The Samuel Gee Lecture 1995. PMID- 8709066 TI - Regional conference in sheffield. Advances and controversies in treatment. PMID- 8709065 TI - Is a pre-discharge checklist useful? AB - A pre-discharge checklist of requirements for equipment, services, benefits and follow-up has been developed in a general rehabilitation unit. An analysis of 66 inpatients discharged after rehabilitation following a stroke or amputation within the past 2 years suggests that such a checklist is a useful tool in ensuring that patients are sent home with optimum services and support. It is also a simple way of recording discharge data for audit and quality control purposes. It may be of benefit to a wider range of hospital services. PMID- 8709067 TI - Medicine and elderly people: over-investigation or under-treatment? PMID- 8709068 TI - The language of eponyms. PMID- 8709069 TI - AIDS in San Francisco. The Medicine Gilliland Travelling Fellowship 1995. PMID- 8709070 TI - The triple queen and her hospital. PMID- 8709071 TI - Aspirin for frail elderly stroke patients? PMID- 8709072 TI - Responsibilities and protection in the use of radioiodine. PMID- 8709073 TI - Advance directives. PMID- 8709074 TI - Babinski or Remak? PMID- 8709075 TI - The earliest known case of a lithopaedion. AB - A lithopaedion, or stone-child, is a dead fetus, usually the result of a primary or secondary abdominal pregnancy, that has been retained by the mother and subsequently calcified. This paper describes the earliest known case of this phenomenon. It was discovered in 1582, at the autopsy of a 68-year-old woman in the French city of Sens, and described in a thesis by the physician Jean d'Ailleboust. The woman had carried her lithopaedion for 28 years. In this historical vignette, the lithopaedion of Sens is compared to later instances of this phenomenon. The ultimate fate of the lithopaedion specimen, which was widely traded throughout Europe in the 1600s before finally ending up in Copenhagen, is traced. PMID- 8709076 TI - Surgery for mitral stenosis in Guy's and the Middlesex Hospitals 1948-1953. PMID- 8709077 TI - Colleges versus academies. PMID- 8709078 TI - The prevalence of eye disease in elderly Bengalis in Tower Hamlets. AB - The prevalence of eye disease and uncorrected refractive errors in a group of 167 elderly members of the Bangladeshi community which resides in the London Borough of Tower Hamlets was studied. Of the subjects screened 24.6% were found to have a significant and potentially treatable cause of visual loss and a further 32.3% were visually handicapped through the presence of uncorrected refractive errors. A high prevalence (53.3%) of cataract was found in the elderly Bengalis. The high prevalence of eye disease in this ethnic minority group, has important implications for health service planning. PMID- 8709079 TI - Dependence of retinopathy (and other complications) on glycaemic control and on weight over 5/10 years from diagnosis of type II diabetes. AB - Glycosylated haemoglobins and weights were recorded for 200 consecutive diabetic clinic attenders seen yearly for 5 years, 76 of whom were also seen up to 10 years from diagnosis of type 2 diabetes, representing 1380 patient years. Weight fluctuation (> 3 kg) was associated with increased final prevalence of hypertension, macroalbuminaemia and a raised creatinine (P < 0.002) but this relationship was abolished by correction for higher initial weight. Average glycaemia over 5/10 years [itself related to initial weight in women on tablets (N = 53) but not others, and to waist but not waist/hip ratio], correlated with prevalence and severity of retinopathy (N = 200; r = 0.38, P < 0.0006) seen also in the subgroup of patients on tablets (N = 145, P < 0.006). At HbA1 levels > 10.5% an increased prevalence of retinopathy was seen in those on insulin (N = 37, P < 0.001) and an increased prevalence of peripheral vascular disease was seen in men but not women (x2 = 2.87, P < 0.01) as well as in the prevalence of neuropathy. These findings suggest that good glycaemic control is of value in type 2 diabetes and less easily achieved in obesity. PMID- 8709080 TI - What is the ultimate cause of socio-economic inequalities in health? An explanation in terms of evolutionary psychology. PMID- 8709081 TI - A multi-observer study examining the radiographic visibility of fishbone foreign bodies. AB - The use of plain radiographs to localize a suspected fishbone foreign body is the subject of controversy. Accordingly radiographs of 14 species of fishbone, impacted in a soft tissue phantom, were assessed by a series of observers from the ENT department (consultant surgeons, senior registrars and house officers). The agreement was assessed by graphical description of the data and tested by a Spearman's rank correlation test. The overall results showed that, for the clinician, radiography is very useful to detect the bones of: cod, haddock, lemon sole, cole fish, grey mullet and plaice; useful for red snapper, monk fish, gurnard and salmon; and unhelpful in detecting bones from herring, pike, mackerel and trout. The use of radiographs to locate these impacted fishbones can be rationalized in the light of these findings. PMID- 8709082 TI - AIDS-related malignancies. AB - The management of AIDS-related malignancies demands that the cancer must be treated in the context of patients already suffering from a fatal disease. Management must be problem-orientated. The standard cancer treatment of such a patient may not be appropriate in the AIDS setting. The AIDS related malignancies are Kaposi's sarcoma (KS) and non-Hodgkin's lymphoma (NHL) including primary cerebral lymphoma. Carcinoma of the cervix was recently added to this list, but has not been seen in this context in the UK. PMID- 8709083 TI - The pentothal test in the management of orthopaedic patients with hysterical illness. AB - Hysterical illness may be defined as symptoms normally associated with disease without pathological basis. Orthopaedics attracts more of these patients than most specialties. They are intensively and expensively investigated, yet clinicians remain anxious they are missing genuine pathology. We use examination under pentothal, without analgesia, to help distinguish real from imagined symptoms. The results of a review of patients evaluated under pentothal over the past two years are presented. PMID- 8709085 TI - ANCA associated with Behcet's disease. AB - Behcet's disease is a multisystem disorder affecting primarily mucocutaneous and ocular sites although the gastrointestinal, cardiovascular, central nervous and respiratory systems may also be involved. Hulusi Behcet, a Turkish dermatologist, first described Behcet's disease in 1937 and suggested a possible infectious aetiology. The pathogenesis of this condition still remains unclear although it is likely that infection acts as a trigger in genetically susceptible individuals. We report a patient with unusual cutaneous manifestations of Behcet's disease and antineutrophil cytoplasmic antibodies (ANCA) directed against the cytotoxic protein, bactericidal/permeability-increasing protein (BPI). This is the first report of Behcet's disease associated with this autoantibody. PMID- 8709084 TI - Bladder exstrophy from childhood into adult life. AB - Exstrophy of the bladder is rare and the incidence of bladder exstrophy is calculated to be from 1 per 30,000 to 50,000 live births with male to female ratio ranging from 1.5-5 to 1(1-4). It was found that persistence or overgrowth of the cloacal membrane on the lower anterior abdominal area, prevents normal mesenchymal ingrowth. This causes divergence of the lower abdominal muscular structures and forces the genital ridges to fuse caudal to the cloacal membrane. The stage of ingrowth of the urorectal septum at the time of rupture determines whether one will produce an exstrophic urinary tract alone (classic bladder exstrophy or epispadias) or cloacal exstrophy with the hindgut interposed between the hemibladders. PMID- 8709086 TI - Favourable outcome of scleroderma renal crisis. AB - Severe hypertension and rapidly progressive acute renal failure is a well recognized complication of scleroderma, often referred to as the renal crisis, and widely thought to cause irreversible deterioration in renal function. With the advent of angiotensin-converting-enzyme inhibitors (ACE-I) the outlook for patients with this condition has dramatically improved. We report here one such patient. PMID- 8709087 TI - A fatal trip with ecstasy: a case of 3,4-methylenedioxymethamphetamine/3,4- methylenedioxyamphetamine toxicity. AB - Since the late 1980s there have been a number of reports about the complications associated with using the designer drug 3,4-methylenedioxymethamphetamine (MDMA) or 'ecstasy'. Although this drug has been widely abused in the past, the potentially fatal complications have arisen recently mainly due to the circumstances of its use at all-night dance sessions or 'raves'. A combination of direct toxicity and strenuous physical exertion can lead to hyperthermia and its attendant complications. The following case illustrates some of the possible consequences following ingestion of 'ecstasy', and reiterates the importance of prompt recognition and treatment in preventing the development of irreversible sequelae. PMID- 8709088 TI - Congenital gingival granular cell tumour. AB - Congenital gingival granular cell tumours are rare lesions which have only occasionally been reported in the UK. Clinical features are of a benign lesion which occurs almost exclusively in newborn, Caucasian females and the anterior maxilla is the commonest site. Treatment consists of local excision and is curative. The terminology concerning this condition has been rather confused because of uncertainty regarding the histogenesis of these tumours and the similar histological appearance to adults granular cell myoblastoma occurring at other intraoral sites. The exact histogenesis of these tumours remains unsolved and they may be hamartomata. We describe a new case occurring within the UK, which illustrates many of the common clinical features of the condition, with an accompanying literature review. PMID- 8709089 TI - Cavernous haemangioma with Kasabach-Merritt syndrome: treatment with alpha interferon. AB - A small number of patients with haemangioma need treatment for serious complications such as Kasabach-Merritt syndrome, cardiac failure and obstruction of the airway. We report on the management of an infant with Kasabach-Merritt syndrome. PMID- 8709090 TI - The death of Mozart. PMID- 8709091 TI - The economics of cardiac failure. AB - Quality assurance and inclusion of prospective evaluation of costs of treatment in phase 3 and 4 pharmaceutical trials are becoming increasingly important. Not only high technology applications have to be investigated, but also relatively cheap but very common strategies for diagnostic work up and therapy. This may yield major savings. We are at the beginning of an era in which waste of resources may be reduced by scientific analysis with improvement in patient care and teaching achieved as a result. PMID- 8709092 TI - Modulators of leukotriene biosynthesis and receptor activation. PMID- 8709093 TI - 3-(1H-indazol-3-ylmethyl)-1,5-benzodiazepines: CCK-A agonists that demonstrate oral activity as satiety agents. PMID- 8709094 TI - Discovery of novel retinoic acid receptor agonists having potent antiproliferative activity in cervical cancer cells. AB - Retinoic acid receptor (RAR) active retinoids have proven therapeutically useful for treating certain cancers and dermatological diseases. Herein, we describe the discovery of two new RAR active trienoic acid retinoids, (2E,4E,6E)-7-(3,5-di tert-butylphenyl)-3-methylocta-2, 4,6-trienoic acid (10a, ALRT1550) and (2E,4E,6Z)-7-(3,5-di-tert-butylphenyl)-3-methylocta-2, 4,6-trienoic acid (10b, LG100567). ALRT1550 is a RAR selective retinoid which exhibits exceptional potency in both competitive binding and cotransfection assays. Moreover, it is the most potent antiproliferative retinoid described to date and thus has implications for the treatment of certain cancers. LG100567 is a potent panagonist which activates both RARs and retinoid X receptors. PMID- 8709095 TI - (S)-13-[(dimethylamino)methyl]-10,11,14,15-tetrahydro-4,9:16, 21-dimetheno-1H, 13H-dibenzo[e,k]pyrrolo[3,4-h][1,4,13]oxadiazacyclohexadecene-1,3(2H)-d ione (LY333531) and related analogues: isozyme selective inhibitors of protein kinase C beta. AB - Protein kinase C (PKC) is a family of closely related serine and threonine kinases. Overactivation of some PKC isozymes has been postulated to occur in several diseases states, including diabetic complications. Selective inhibition of overactivated PKC isozymes may offer a unique therapeutic approach to disease states such as diabetic retinopathy. A novel series of 14-membered macrocycles containing a N-N'-bridged bisindolylmaleimide moiety is described. A panel of eight cloned human PKC isozymes (alpha, beta I, beta II, gamma, delta, epsilon, sigma, eta) was used to identify the series and optimize the structure and associated activity relationship. The dimethylamine analogue LY333531 (1), (S)-13 [(dimethylamino)methyl]-10,11,14,15-tetrahydro-4,9:16, 21-dimetheno-1H, 13H dibenzo[e,k]pyrrolo[3,4-h][1,4,13]oxadiazacyclohexadecene++ +-1,3(2H)-dione, inhibits the PKC beta I (IC50 = 4.7 nM) and PKC beta II (IC50 = 5.9 nM) isozymes and was 76- and 61-fold selective for inhibition of PKC beta I and PKC beta II in comparison to PKC alpha, respectively. The additional analogues described in the series are also selective inhibitors of PKC beta. LY333531 (1) exhibits ATP dependent competitive inhibition of PKC beta I and is selective for PKC in comparison to other ATP dependent kinases (protein kinase A, calcium calmodulin, caesin kinase, src tyrosine kinase). The cellular activity of the series was assessed using bovine retinal capillary endothelial cells. Retinal endothelial cell dysfunction has been implicated in the development of diabetic retinopathy. Plasminogen activator activity stimulated by a phorbol ester (4 beta-phorbol 12,13-dibutyrate) in endothelial cells was inhibited by the compounds in the series with ED50 values ranging from 7.5 to 0.21 microM. A comparison of the PKC isozyme and related ATP dependent kinase inhibition profiles is provided for the series and compared to the profile for staurosporine, a nonselective PKC inhibitor. The cellular activity of the series is compared with that of the kinase inhibitor staurosporine. PMID- 8709096 TI - Pyrrolobenzothiazepinones and pyrrolobenzoxazepinones: novel and specific non nucleoside HIV-1 reverse transcriptase inhibitors with antiviral activity. AB - Two novel classes of pyrrolobenzothiazepinones and pyrrolobenzoxazepinones were investigated as potential anti-AIDS drugs. These compounds were found to inhibit HIV-1 reverse transcriptase (RT) enzyme in vitro and to prevent HIV-1 cytopathogenicity in T4 lymphocytes, without appreciable activity on HIV-2 cytopathic effects, and against HBV as well as calfthymus DNA alpha-polymerase. Their potency is influenced by substituents at position 6 and on the fused aromatic ring. Specifically, small lipophilic substituents at C-6 were preferred, whereas substitutions on the benzo-fused ring were found to be detrimental to activity, with respect to the unsubstituted compounds. Modification of the pie system at C-6 is well tolerated, although the replacement of the benzo-fused with a [2,3]naphtho-fused ring leads to a less active compound. Maximum potency and specificity is achieved with a phenyl and an ethyl group at position 6 of the pyrrolobenzoxazepinone system. In the enzymatic assay the oxazepinone derivative (+/-)-6-ethyl-6-phenylpyrrolo[2,1-d][1,5] benzoxazepin-7(6H)-one 16e (IC50 = 0.25 microM) was found to be more potent than nevirapine (IC50 = 0.5 microM), tested in the same experimental conditions using rC.dG as a template-primer. In cell culture assay benzoxazepine 16e was active against HIV-1, both wild type and AZT sensitive, and HIV-1 (IIIB) strains, but not against HIV-2. In enzyme assay although 16e inhibited HIV-1 RT, it was inactive against the nevirapine-resistant recombinant RT Y181C at 50 microM. Molecular modeling studies suggest that these derivatives present a 3D pharmacophoric arrangement similar to that of other non nucleoside inhibitors such as nevirapine. PMID- 8709097 TI - Inhibitors of human immunodeficiency virus type 1 derived from gp41 transmembrane protein: structure--activity studies. AB - We synthesized analogues of gp41 (553-590), 1, and evaluated them for their inhibitory activity against HIV-1 in MT4 cell assay (IC50(1) = 2.7 microM). (The numbering scheme for gp41 (e.g., gp41(553-590) for 1) adapted throughout the text is from ref 6.) Gradual truncation of either the N- or C-terminal end of gp41 (553-590) resulted in a substantial loss of inhibitory properties of resulting compounds. Unexpectedly, simultaneous truncations of both N- and C-termini of gp41(553-590) resulted in a potent heptadecamer, 13, IC50 = 10.4 microM. Coupling of a racemic alpha-aminotetradecanoic acid (Atd) to gp41 fragments afforded diastereomeric conjugates, most of which were chromatographically separable. In this series, pentadecamer 27 had an IC50 of 8.9 microM, while its Atd diastereomer 28 was much less inhibitory. This finding is consistent with relative inhibitory potencies of other Atd-containing diastereomeric pairs and could reflect a chiral sense of Atd residue interacting with the receptor. Compounds 13 and 27, which are practically equipotent to 1, represent minimalistic fragments of the leucine-zipper region of gp41 and constitute a basis for design of a second generation of gp41-based inhibitors. Circular dichroism studies suggested that compounds in this series are likely to inhibit HIV-1 replication by virtue of their alpha-helical character. The observed structure-activity relationship supports impairment of viral gp41 as a possible mechanism of action of 1. PMID- 8709098 TI - Rational design of novel immunosuppressive drugs: analogues of azathioprine lacking the 6-mercaptopurine substituent retain or have enhanced immunosuppressive effects. AB - Clinical use of the immunosuppressive drug azathioprine is limited by potentially serious toxic effects related to depression of bone marrow function. The immunosuppressive and toxic properties of azathioprine are regarded as being properties of the cytotoxicity of its metabolite, 6-mercaptopurine (6-MP). However, azathioprine has an immunosuppressive effect additional to that attributable to 6-MP alone, and we propose that this is associated with an action of the methylnitroimidazolyl substituent. This suggests a route to the rational design of nontoxic immunosuppressants by replacing the 6-MP component of azathioprine with nontoxic thiols. We have synthesized and tested in vitro 24 such analogues, with two being further tested in vivo. In the human mixed lymphocyte reaction, virtually all compounds showed some degree of activity, 10 compounds being more active than azathioprine. In vivo, two compounds were more effective than azathioprine at prolonging graft survival in mice. In an oral toxicity study in male CD1 mice at doses equivalent to those at which azathioprine caused severe bone marrow depression both analogues had no toxic effects. Our results show that the immunosuppressive effects and bone marrow toxicity of azathioprine are not a consequence of release of 6-MP alone, and with appropriate modification can be separated, an approach which may lead to less toxic immunosuppressive drugs. PMID- 8709099 TI - Novel selective PDE IV inhibitors as antiasthmatic agents. Synthesis and biological activities of a series of 1-aryl-2,3-bis(hydroxymethyl)naphthalene lignans. AB - A series of 1-aryl-2,3-bis(hydroxymethyl)naphthalene lignans have been synthesized and evaluated for their ability to selectively inhibit PDE IV isolated from guinea pig. Replacement of the 1-phenyl ring by a pyridone ring led to marked improvement of their selectivity for PDE IV over PDE III. The compounds that were most potent and selective involved those bearing an N-alkylpyridone ring at C-1. These compounds also showed potent antispasmogenic activity without causing significant changes in heart rate in the guinea pig. The most potent compound was 6,7-diethoxy-2, 3-bis(hydroxymethyl)-1-[1-(2-methoxyethyl)-2-oxo pyrid-4-yl]nap hth alene (17f), ED50 values of histamine-induced and antigen induced bronchoconstriction in the guinea pig being 0.08 and 2.3 mg/kg iv, respectively. This compound was chosen as a candidate for further pharmacological evaluation. PMID- 8709100 TI - Preparation of phenolic paclitaxel metabolites. AB - The synthesis and biological evaluation of the two known phenolic metabolites of paclitaxel are described. The C3'-phenolic metabolite 2 of paclitaxel was prepared from 7-(triethylsilyl)-baccatin III (8) and enantioenriched N-benzoyl-2 azetidinone 7. The C2-phenolic metabolite 3 was synthesized from paclitaxel (1a) via selective C2 debenzoylation and reacylation. PMID- 8709101 TI - Deconvolution of combinatorial libraries for Drug discovery: theoretical comparison of pooling strategies. AB - Synthesis and testing of mixtures of compounds in a combinatorial library allow much greater throughput than synthesis and testing of individual compounds. When mixtures of compounds are screened, however, the possibility exists that the most active compound will not be identified. The specific strategies employed for pooling and deconvolution will affect the likelihood of success. We have used a nucleic acid hybridization example to develop a theoretical model of library deconvolution for a library of more than 250,000 compounds. This model was used to compare various strategies for pooling and deconvolution. Simulations were performed in the absence and presence of experimental error. We found iterative deconvolution to be most reliable when active molecules were assigned to the same subset in early rounds. Reliability was reduced only slightly when active molecules were assigned randomly to all subsets. Iterative deconvolution with as many as 65,536 compounds per subset did not drastically reduce the reliability compared to one-at-a-time testing. Pooling strategies compared using this theoretical model are compared experimentally in an accompanying paper. PMID- 8709102 TI - Deconvolution of combinatorial libraries for drug discovery: experimental comparison of pooling strategies. AB - An experimental evaluation of several different pooling strategies for combinatorial libraries was conducted using a library of 810 compounds and an enzyme inhibition assay (phospholipase A2). The library contained compounds with varying degrees of activity as well as inactive compounds. The compounds were synthesized in groups of three and pooled together in various formats to realize different pooling strategies. With one exception, all iterative deconvolution strategies and position scanning resulted in identification of the same compound. The results are in good agreement with the predicted outcome from theoretical and computational methods. These data support the tenet that active compounds for pharmaceutically relevant targets can be successfully identified from combinatorial libraries organized in mixtures. PMID- 8709103 TI - Conformationally restricted analogs of 1 alpha, 25-dihydroxyvitamin D3 and its 20 epimer: compounds for study of the three-dimensional structure of vitamin D responsible for binding to the receptor. AB - Two proteins play important roles in the expression of vitamin D function: the specific nuclear receptor protein (vitamin D receptor, VDR) and the transport protein (vitamin D binding protein, DBP). This study was conducted to clarify the conformation of vitamin D responsible for binding to those proteins. For the purpose, the side chain mobility of 1,25(OH)2D3 (1) and its 20-epimer, 20-epi 1,25(OH)2D3 (2), was analyzed by a systematic conformational search. The results were depicted as a three-dimensional dot map, which indicates that the side chains of the two vitamins (1 and 2) occupy different spatial regions that are separated in two areas. We denoted these areas as A and G for 1 and EA and EG for 2. Four analogs, the diastereomers at C(20) and C(22) (3-6) of 22-methylated 1,25(OH)2D3 whose side chains were confined to occupy G, A, EA, and EG, respectively, were designed. These analogs (3-6) were synthesized efficiently by a stereoselective conjugate addition of organocuprate to steroidal E- and Z-22-en 24-ones as the key step. In binding to the VDR the affinities of the analogs (3 6) relative to 1,25-(OH)2D3 (1) were 1/60, 1/3, 20, and 1/100, respectively. These results indicate that the A region is responsible for binding of 1 to VDR and the EA region for binding of 2. Only isomer 4 showed significant affinity for DBP, indicating only the A region is responsible for binding to DBP. Thus, 5 showed clear separation of binding affinities for two proteins, VDR and DBP. Having the highest known VDR affinity, (22R)-22-methyl-20-epi-1,25(OH)2D3 (5) has potential both as a therapeutic agent and as a tool to study the molecular mechanism of vitamin D-mediated gene transcription. PMID- 8709105 TI - Forskolin carbamates: binding and activation studies with type I adenylyl cyclase. AB - Three series of analogs were regioselectively prepared from a protected forskolin precursor to afford 7-carbamoyl-7-desacetylforskolins (series 1), 6-carbamoyl-7 desacetylforskolins (series 2), and 6-carbamoylforskolins (series 3). The analogs were pharmacologically evaluated for binding (IC50) to and activation (EC50) of type I adenylyl cyclase in membranes from stably transfected Sf9 cell lines expressing a single adenylate cyclase subtype. The following ranges were determined for the IC50's and EC50's of each individual series: series 1, IC50 = 43-1600 nM, EC50 = 0.5-9.6 microM; series 2, IC50 = 65-680 nM, EC50 = 0.63-6.5 microM; series 3, IC50 = 21-271 nM, EC50 = 0.5-8.1 microM (forskolin IC50 = 41 nM and EC50 = 0.5 microM). Activation paralleled binding; however, some analogs exhibited poor binding and good activation whereas others demonstrated good binding but poor activation. Steric bulk tended to diminish binding and activation when at the 6- or 7-position, although bulk was accommodated at the 6 position if the 7-site was reacetylated. Acylation of the 7-position by the carbamoyl linker or acetyl was important for obtaining good binding and activation; however, the effect was more pronounced with binding. For both binding and activation, small, linear, lipophilic substituents (propyl, allyl, isopropyl) are well tolerated at the 7-position but less so in the 6-position, even when the 7-site is reacetylated. Planar aromatic moieties (phenyl and 2 pyridinyl) demonstrated moderate to good potency for binding and activation when located at either the 6- or 7-positions. There is an overall trend toward increasing potency for both binding and activation with polar substituents. PMID- 8709104 TI - Novel cyclic analogs of angiotensin II with cyclization between positions 5 and 7: conformational and biological implications. AB - To study the conformational features of molecular recognition of angiotensin II (Asp1-Arg2-Val3-Tyr4-Val/IIe5-His6-Pro7-Phe8, AII), the synthesis and biological testing of several cyclic analogs of AII cyclized between positions 5 and 7 have been performed. The synthesized analogs were Sar1-Arg2-Val3-Tyr4-cyclo(Cys5-His6 Pen7)-Phe8 (3), Sar1-Arg2-Val3-Tyr4-cyclo(Asp5-His6-Apt7)-Phe8 (4), Sar1-Arg2 Val3-Tyr4-cyclo(Glu5-His6-Apt7)-Phe8 (5), Sar1-Arg2-Val3-Tyr4-cyclo-(Cys5-His6 Mpt7)-Phe8 (6), Sar1-Arg2-Val3-Tyr4-cyclo(Cys5-His6-Mpc7)-Phe8 (7), Sar1-Arg2 Val3-Tyr4-cyclo(Hcy5-His6-Mpt7)-Phe8 (8), and Sar1-Arg2-Val3-Tyr4-cyclo(Hcy5-His6 Mpc7)-Phe8 (9), where Apt stands for 4-amino-trans-proline, and Mpt and Mpc for 4 mercapto-trans- and -cis-prolines, respectively. Compound (9) showed good affinity at AT-1 receptors, namely a KD = 20 nM. In functional assays, it showed the characteristics of a weak partial agonist with a relative affinity of 0.26% of that for AII and an intrinsic efficacy, alpha E, of 0.42. Molecular modeling suggested a possible explanation for this finding: the relatively strong binding and the weak partial agonistic activity of compound 9 are due to interaction with AT-1 receptor of only two functionally important groups, namely, the side chains of the His6 and Phe8 residues. PMID- 8709106 TI - Synthesis, ligand binding, and quantitative structure-activity relationship study of 3 beta-(4'-substituted phenyl)-2 beta-heterocyclic tropanes: evidence for an electrostatic interaction at the 2 beta-position. AB - A set of 3 beta-(4'-substituted phenyl)-2 beta-heterocyclic tropanes was designed, synthesized, and characterized. We discovered that these compounds can function as bioisosteric replacements for the corresponding WIN 35,065-2 analogs which possess a 2 beta-carbomethoxy group. Several of the compounds showed high affinity and selectivity for the dopamine transporter (DAT) relative to the serotonin and norepinephrine transporters. From the structure-activity relationship study, the 3 beta-(4'-chlorophenyl)-2 beta-(3'-phenylisoxazol-5 yl)tropane (5d) emerged as the most potent and selective compound. The binding data for 2 beta-heterocyclic tropanes were found to show a high correlation with molecular electrostatic potential (MEP) minima near one of the heteroatoms in the 2 beta-substituents. In contrast, low correlations were found for other MEP minima near the 2 beta-substituent as well as for calculated log P or substituent volume. These quantitative structure-activity relationship studies are consistent with an electrostatic contribution to the binding potency of these WIN 35,065-2 analogs at the DAT. PMID- 8709107 TI - Dopamine D3 and D4 receptor antagonists: synthesis and structure--activity relationships of (S)-(+)-N-(1-Benzyl-3-pyrrolidinyl)-5-chloro-4- [(cyclopropylcarbonyl) amino]-2-methoxybenzamide (YM-43611) and related compounds. AB - In this study, we synthesized a series of (S)-N-(3-pyrrolidinyl)benzamide derivatives, 1, 2a-d, 5a-1, and 7, and their enantiomers, (R)-1 and (R)-5c-e, and evaluated their binding affinity for cloned dopamine D2, D3, and D4 receptors and their inhibitory activity against apomorphine-induced climbing behavior in mice. The results indicate that D2, D3, and D4 receptors have different bulk tolerance (D4 > D3 > D2) for the substituent of the 4-amino group (R1) on the benzamide nuclei and that cyclopropyl-, cyclobutyl-, and cyclopentylcarbonyl groups likely possess adequate bulkiness with respect to D3 and D4 affinity and selectivity over D2 receptors in this series. The results also suggested that the N substituent (R2) on the pyrrolidin-3-yl group performs an important role in expressing affinity for D2, D3, and D4 receptors and selectivity among the respective subtypes. One of the compounds, (S)-(+)-N-(1-benzyl-3-pyrrolidinyl)-5 chloro-4-[(cyclopropylcarbonyl+ ++) amino]-2-methoxybenzamide (5c) (YM-43611), showed high affinity for D3 and D4 receptors (Ki values of 21 and 2.1 nM, respectively) with 110-fold D4 selectivity and 10-fold D3 preference over D2 receptors and weak or negligible affinity for representative neurotransmitter receptors. Compound 5c displayed potent antipsychotic activity in inhibiting apomorphine-induced climbing behavior in mice (ED50 value, 0.32 mg/kg sc). PMID- 8709108 TI - Potent, selective tetrahydro-beta-carboline antagonists of the serotonin 2B (5HT2B) contractile receptor in the rat stomach fundus. AB - A series of potent, selective 5HT2B receptor antagonists has been identified based upon yohimbine, with SAR studies resulting in a 1000-fold increase in 5HT2B receptor affinity relative to the starting structure (-log KBS > 10.0 have been obtained). These high-affinity tetrahydro-beta-carboline antagonists are able to discriminate among the 5HT2 family of serotonin receptors, with members of the series showing selectivities of more than 100-fold versus both the 5HT2A and 5HT2C receptors based upon radioligand binding and functional assays. As the first compounds reported with such selectivity and enhanced receptor affinity, these tetrahydro-beta-carboline antagonists are useful tools for elucidating the role of serotonin acting at the 5HT2B receptor in normal and disease physiology. PMID- 8709109 TI - Structure-based design and synthesis of substituted 2-butanols as nonpeptidic inhibitors of HIV protease: secondary amide series. AB - The design, synthesis, and crystallographic analysis of protein-inhibitor complexes is described for a novel series of nonpeptidic HIV protease (HIV Pr)inhibitors. Beginning with a cocrystal structure of a Phe-Pro peptidomimetic bound to the HIV Pr, design was initiated that resulted in the substituted 2 butanol compound 8 as the lead compound (Ki = 24.5 microM, racemic mixture). Modifications on the initial compound were then made on the basis of its cocrystal structure with HIV Pr and inhibition data, resulting in compounds with enhanced potency against the enzyme (compound 18, Ki = 0.48 microM). These inhibitors were found to bind to the enzyme essentially as predicted on the basis of the original design hypothesis. Stereospecific synthesis of individual enantiomers confirmed the prediction of a binding preference for the S alcohol stereochemistry. Modest antiviral activity was demonstrated for several of the more potent HIV Pr inhibitors in a HIV-1 infected CEM-SS cell line. PMID- 8709110 TI - Bis tertiary amide inhibitors of the HIV-1 protease generated via protein structure-based iterative design. AB - A series of potent nonpeptide inhibitors of the HIV protease have been identified. Using the structure of compound 3 bound to the HIV protease, bis tertiary amide inhibitor 9 was designed and prepared. Compound 9 was found to be about 17 times more potent than 3, and the structure of the protein-ligand complex of 9 revealed the inhibitor binds in an inverted binding mode relative to 3. Examination of the protein-ligand complex of 9 suggested several modifications in the P1 and P1' pockets. Through these modifications it was possible to improve the activity of the inhibitors another 100-fold, highlighting the utility of crystallographic feedback in inhibitor design. These compounds were found to have good antiviral activity in cell culture, were selective for the HIV protease, and were orally available in three animal models. PMID- 8709111 TI - Cyclopent[a]anthraquinones as DNA intercalating agents with covalent bond formation potential: synthesis and biological activity. AB - A series of mitomycin C (MMC) analogues, namely cyclopentanthraquinone derivatives, were synthesized via Diels-Alder cyclization of naphthoquinone with 1-vinylcyclopent-1-enes. These new compounds are planar structures, like MMC, and bear an aziridine ring and a methyl carbamate side chain. After bioreduction, they are anticipated to be capable of intercalating into double-stranded DNA and bind covalently. Structure-activity relationships were studied. Of these compounds, 2,3-aziridino-4-[[(methylamino)carbonyl]methyl] cyclopent[alpha]anthracene-6,11-dione (4) was shown to have inhibitory activity against several leukemic and solid tumor cell lines. Mice (BDF1) bearing Lewis lung adenocarcinoma were treated with 4 and MMC (i.p., QD x 5). At a dose of 30.0 mg/kg, compound 4 was as effective as MMC (0.8 mg/kg). Compound 4 appears to be less toxic than MMC. DNA unwinding assay indicated that 4 is able to intercalate into DNA double strands and is also a topoisomerase II inhibitor. PMID- 8709113 TI - Structure--activity relationships of 1-(2-Deoxy-2-fluoro-beta-L arabinofuranosyl)pyrimidine nucleosides as anti-hepatitis B virus agents. AB - Since 2'-fluoro-5-methyl-beta-L-arabinofuranosyluracil (L-FMAU) has been shown to be a potent anti-HBV agent in vitro, it was of interest to study the structure activity relationships of related nucleosides. Thus, a series of 1-(2-deoxy-2 fluoro-beta-L-arabinofuranosyl)pyrimidine nucleosides have been synthesized and evaluated for antiviral activity against HBV in 2.2.15 cells. For this study, L ribose was initially used as the starting material. Due to the commercial cost of L-ribose, we have developed an efficient procedure for the preparation of L ribose derivative 6. Starting from L-xylose, 6 was obtained in an excellent total yield (70%) through the pyridinium dichromate oxidation of the 3-OH group followed by stereoselective reduction with NaBH4. It was further converted to the 1,3,5-tri-O-benzoyl-2-deoxy-2-fluoro-alpha-L-arabinofuranose (10), which was then condensed with various 5-substituted pyrimidine bases to give the nucleosides. Among the compounds synthesized, the lead compound, L-FMAU (13), exhibited the most potent anti-HBV activity (EC50 0.1 microM). None of the other uracil derivatives showed significant anti-HBV activity up to 10 microM. Among the cytosine analogues, the cytosine (27) and 5-iodocytosine (35) derivatives showed moderately potent anti-HBV activity (EC50 1.4 and 5 microM, respectively). The cytotoxicity of these nucleoside analogues has also been assessed in 2.2.15 cells as well as CEM cells. None of these compounds displayed any toxicity up to 200 microM in 2.2.15 cells. Thus, compound 13 (L-FMAU), 27, and 35 showed a selectivity of over 2000, 140, and 40, respectively. PMID- 8709112 TI - Structure--activity relationships of the didemnins. AB - Bioactivities of 42 didemnin congeners, either isolated from the marine tunicates Trididemnun solidum and Aplidium albicans or prepared synthetically and semisynthetically, have been compared. The growth inhibition of various murine and human tumor cells and plaque reduction of HSV-1 and VSV grown on cultured mammalian cells were used to assess cytotoxicity and antiviral activity. Biochemical assays for macromolecular synthesis (protein, DNA, and RNA) and enzyme inhibition (dihydrofolate reductase, thymidylate synthase, DNA polymerase, RNA polymerase, and topoisomerases I and II) were also performed to specify the mechanisms of action of each analogue. Immunosuppressive activity of the didemnins was determined using a mixed lymphocyte reaction (MLR) assay. These assays revealed that the native cyclic depsipeptide core is an essential structural requirement for most of the bioactivites of the didemnins, especially for cytotoxicities and antiviral activities. The linear side-chain portion of the peptide can be altered with a gain, in some cases, of bioactivities. In particular, dehydrodidemnin B, tested against several types of tumor cells and in in vivo studies in mice, as well as didemnin M, tested for the mixed lymphocyte reaction and graft vs host reaction in murine systems, showed remarkable gains in their in vitro and in vivo activities compared to didemnin B. PMID- 8709114 TI - Synthesis and structure--activity relationships of fused imidazopyridines: a new series of benzodiazepine receptor ligands. AB - 2-Arylimidazo[4,5-c]quinolines and analogous fused imidazopyridines were synthesized and evaluated as benzodiazepine receptor ligands. Affinity to the receptors was greatly affected by the bulkiness of the aryl group at the 2 position, compared to the pyrazoloquinolines such as CGS-9896. Derivatives with an isoxazole moiety at the 2-position showed high binding affinity and in vivo activity. In the imidazo[4,5-c]quinoline series, substitution at the 6-position decreased or abolished activity. Most derivatives with an unsubstituted isoxazolyl group showed antagonist or inverse agonist activity except for the 7 halo analogues, which exhibited agonist activity. On the other hand, 5 methylisoxazol-3-yl or 3-methylisoxazol-5-yl derivatives generally exhibited agonist activity. A similar substitution effect on the isoxazole moiety was observed in the imidazopyridines fused with a nonaromatic ring. From the detailed pharmacological evaluation, S-8510, 2-(3-isoxazolyl)-3,6,7,9 tetrahydroimidazo[4,5-d]pyrano++ +[4,3-b]pyridine monophosphate, possessing weak inverse agonist activity was selected as a therapeutic candidate for the treatment of some symptoms of senile dementia. PMID- 8709115 TI - Potential antitumor agents. 24. Synthesis and pharmacological behavior of imidazo[2,1-b]thiazole guanylhydrazones bearing at least one chlorine. AB - In connection with a previous research dealing with the antitumor activity of imidazo[2,1-b]-thiazole guanylhydrazones, this paper reports the synthesis of new derivatives which were tested for antitumor and positive inotropic activity. In most cases the cytotoxic data from the in vitro experiments (HeLa) were in agreement with the antitumor data in vivo (Ehrlich). The active compounds bear a phenyl ring at the 6 position. On the other hand, the most active cardiotonic agents were devoid of the phenyl ring. PMID- 8709116 TI - Synthesis of acyclo-C-nucleosides in the imidazo[1,2-a]pyridine and pyrimidine series as antiviral agents. AB - The synthesis and the antiviral activities of C-3 acyclic nucleoside analogues of imidazo[1,2-a]pyridine and pyrimidine are reported. From these compounds, 20, 21, 22, 23, 28, and 34 showed a specific activity against cytomegalovirus and/or varicella-zoster virus. PMID- 8709117 TI - New steroidal diazo ketones as potential photoaffinity labeling reagents for the mineralocorticoid receptor: synthesis and biological activities. AB - Three diazo ketones in the progesterone series were synthesized as potential photoaffinity reagents. The diazo ketone group was introduced at the C17 (21 diazopregn-4-ene-3,20-dione, 1) or C13 (18-(diazomethyl)-20-hydroxypregn-4-ene 3,18-dione, 2, 18-(diazomethyl)pregn-4-ene-3, 18,20-trione, 3) position of the pregnene skeleton. Whereas compound 1 could be easily obtained from the corresponding acid chloride, preparation of 2 and 3 required a less straightforward route involving reaction of tosyl azide on the formyl derivative of methyl ketone 5. The affinity of the diazo ketones for the human mineralocorticoid receptor (hMR), expressed in Sf9 insect cells using the Baculovirus system, was estimated by competition experiments using [3H]aldosterone as specific ligand. The affinity of 1 for hMR was almost identical with that of aldosterone. The affinities of 2 and 3 were 1, order of magnitude lower than that of aldosterone. The mineralocorticoid activity of the diazo ketones was measured in cis-trans cotransfection assays in CV-1 cells with the mouse mammary tumor virus as DNA target sequence. Compound 1 exhibits an agonist activity (ED50 = 6 x 10(-9) M) with no antagonist activity. In contrast 2 and 3 behave as antagonists, displaying an IC50 of approximately 10(-6) M whether the substituent at the C20 position is a hydroxy (2) or an oxo (3) group. PMID- 8709118 TI - Immunosuppressive cyclolignans. AB - The immunosuppressive activity of several lactonic, nonlactonic, and heterocycle fused cyclolignans has been demonstrated for the first time by use of a T-cell mediated immune response. Of the compounds tested, 4' demethyldeoxypodophyllotoxin (8), beta-apopicropodophyllin (6), and the isoxazoline-fused cyclolignan 15 are the most potent with respect to their suppression of activated splenocytes. PMID- 8709119 TI - (2E)-5-[3-[(phenylsulfonyl)amino]phenyl]-pent-2-en-4-ynohydroxamic acid and its derivatives as novel and potent inhibitors of ras transformation. PMID- 8709120 TI - (S)-(+)-2-(3'-carboxybicyclo[1.1.1]pentyl)-glycine, a structurally new group I metabotropic glutamate receptor antagonist. PMID- 8709121 TI - Conformational studies and electronic structures of tamoxifen and toremifene and their allylic carbocations proposed as reactive intermediates leading to DNA adduct formation. AB - Toremifene, a compound which differs from tamoxifen by the substitution of a chlorine atom for a hydrogen atom in the ethyl group, is significantly less potent than tamoxifen in causing DNA adduct formation in rats. To examine the relationship of the DNA adduct-forming ability of these compounds with their physicochemical properties such as stable conformation and chemical reactivity, we carried out molecular mechanics, molecular dynamics, and quantum mechanics calculations for the two compounds. For tamoxifen, six stable conformers were identified by conformational search with CFF91 force field. Molecular dynamics simulations showed that these were often interconverted within 1.0 ns. On the other hand, although the conformation of stable conformers and dynamical behavior of toremifene were almost the same as those of tamoxifen, a few conformations were slightly different from those of tamoxifen owing to the effect of the chlorine atom at chloroethyl group. In addition, the stability of the allylic carbocation, which had been proposed as the reactive intermediate leading to DNA adduct formation, was calculated with both semiempirical and density functional methods. Results showed that the carbocation intermediate of toremifene was less stable than that of tamoxifen by 4-5 kcal/mol, suggesting that toremifene was less frequently activated to the intermediate than tamoxifen. Furthermore, the carbocation intermediates of two other tamoxifen derivatives, 4-iodotamoxifen and droxifene, which show no DNA adduct-forming ability, were also less stable compared with that of tamoxifen. These calculated results suggest a close relation between the stability of the proposed carbocation intermediate and DNA adduct-forming ability. PMID- 8709122 TI - The properties of known drugs. 1. Molecular frameworks. AB - In order to better understand the common features present in drug molecules, we use shape description methods to analyze a database of commercially available drugs and prepare a list of common drug shapes. A useful way of organizing this structural data is to group the atoms of each drug molecule into ring, linker, framework, and side chain atoms. On the basis of the two-dimensional molecular structures (without regard to atom type, hybridization, and bond order), there are 1179 different frameworks among the 5120 compounds analyzed. However, the shapes of half of the drugs in the database are described by the 32 most frequently occurring frameworks. This suggests that the diversity of shapes in the set of known drugs is extremely low. In our second method of analysis, in which atom type, hybridization, and bond order are considered, more diversity is seen; there are 2506 different frameworks among the 5120 compounds in the database, and the most frequently occurring 42 frameworks account for only one fourth of the drugs. We discuss the possible interpretations of these findings and the way they may be used to guide future drug discovery research. PMID- 8709123 TI - Inhibition of UDP-N-acetylglucosamine import into Golgi membranes by nucleoside monophosphates. AB - The specificity of the UDP-N-acetylglucosamine (UDP-GlcNAc) translocator for the binding of nucleoside monophosphates (NMPs) and nucleotide-sugars was examined in order to develop a quantitative understanding of how this enzyme recognizes its substrates and to provide a framework for development of novel drugs that target glycosylation. Competition studies reveal that tight binding requires a complete ribose ring and a 5'-phosphate. The enzyme is extremely tolerant to changes at the 3'-position, and the presence of 3'-F actually increases binding of the NMP to the enzyme. At the 2'-position, substitutions in the ribo configuration are well tolerated, although these same substitutions greatly diminish binding when present in the ara configuration. For the base, size appears to be the key feature for discrimination. The enzyme tolerates changing the C-4 oxygen of uridine to an amino group as well as substituting groups containing one or two carbons at C-5. However, substitution of groups containing three carbons at C-5, or exchange of the pyrimidine for a purine, greatly weakens binding to the translocator. Comparison of various UDP-sugars reveals that the UDP-GlcNAc translocator has lower affinity for UDP-N-acetylgalactosamine and UDP-glucose than for its cognate substrate and therefore indicates that this translocator requires both proper stereochemistry at C-4 and an aminoacetyl group at C-2. The impact of these observations on the design of more powerful nucleoside-based inhibitors of nucleotide-sugar import is discussed. PMID- 8709124 TI - Structure-activity relationships of the antimalarial agent artemisinin. 4. Effect of substitution at C-3. AB - Novel antimalarial artemisinin analogs, 3-alkylartemisinins as well as 3 (arylalkyl)- and 3-(carboxyalkyl)artemisinins, were prepared via the synthetic intermediate 2. Formation of the N,N-dimethylhydrazones 5 and 24 and then regio- and chemoselective deprotonation followed by alkylation provided initially alkylated hydrazones that upon chromatography gave ketones 6-13 and 25-30. Direct ozonolysis of the ketones followed by in situ acidification lead directly to the formation of title compounds 14-21 and 31-36. The analogs were tested in vitro against W-2 and D-6 strains of Plasmodium falciparum and found to be in some cases much more active than the natural product (+)-artemisinin. The results were included in structure-activity relationship (CoMFA) studies for further analog design. PMID- 8709125 TI - N-heteroaryl-2-phenyl-3-(benzyloxy)piperidines: a novel class of potent orally active human NK1 antagonists. AB - The preparation of a series of N-heteroarylpiperidine ether-based human NK1 antagonists is described. Two of the compounds 3-[-(2S,3S)-3-(((3,5 bis(trifluoromethyl)phenyl)methyl)oxy)- 2-phenylpiperidino}methyl]-1,2,4-triazole (11) and 5-[?(2S,3S)-3-(((3,5-bis(trifluoromethyl)-phenyl)methyl)oxy)-2- phenylpiperidino}methyl]-3-oxo-1,2,4-triazolone (12)), in particular, are orally bioavailable and exhibited significant improvements in potency, both in vitro and in vivo, over the lead (carboxamidomethyl)piperidine ether 1. Rat liver microsome studies on a selected number of compounds from this series show the triazolone heterocycle to be considerably more stable than the others. Furthermore, both 11 and 12 have been profiled in a number of assays that may be predictive of the clinical utility of substance P antagonists. PMID- 8709126 TI - Synthesis and binding activity of some pyrazolo[1,5-c]quinazolines as tools to verify an optional binding site of a benzodiazepine receptor ligand. AB - The synthesis and binding activity at the benzodiazepine receptor of some 2 substituted pyrazolo[1,5-c]quinazolines are reported. The structure-activity relationships and in vitro efficacy of the title compounds, which are devoid of the proton acceptor atom at position 1, are similar to those of some previously reported tricyclic heteroaromatic compounds. This suggests that a proton acceptor at position 1 is an optional binding site of a benzodiazepine receptor ligand which only affects potency. PMID- 8709127 TI - Cardiovascular characterization of pyrrolo[2,1-d][1,5]benzothiazepine derivatives binding selectively to the peripheral-type benzodiazepine receptor (PBR): from dual PBR affinity and calcium antagonist activity to novel and selective calcium entry blockers. AB - The synthesis and cardiovascular characterization of a series of novel pyrrolo[2,1-d][1,5]-benzothiazepine derivatives (54-68) are described. Selective peripheral-type benzodiazepine receptor (PBR) ligands, such as PK 11195 and Ro 5 4864, have recently been found to possess low but significant inhibitory activity of L-type calcium channels, and this property is implicated in the cardiovascular effects observed with these compounds. In functional studies both PK 11195 (1-(2 chlorophenyl)-N-methyl-N-(1-methylpropyl)-3-isoquinolinecarboxa mide) and Ro 5 4864 (4'-chlorodiazepam) did not display selectivity between cardiac and vascular tissue. Therefore, several 7-(acyloxy)-6-arylpyrrolo[2,1-d][1,5]benzothiazepines, potent and selective peripheral-type benzodiazepine receptor ligands recently developed by us (3, 7-20), were subjected to calcium channel receptor binding assay. Some of these compounds showed an unexpected potency in displacing the binding of [3H]nitrendipine from L-type calcium channels, much higher than that reported for PK 11195 and Ro 5-4864 and equal to or higher than that of reference calcium antagonists such as verapamil and (+)-cis-diltiazem. Specifically, in rat cortex homogenate, our prototypic PBR ligand 7-acetoxy-6-(p methoxyphenyl)pyrrolo[2,1-d][1,5]benzothiazepine (3) showed an IC50 equal to 0.13 nM for inhibition of [3H]nitrendipine binding. Furthermore, in functional studies this compound displayed a clear-cut selectivity for cardiac over vascular tissue. Comparison of calcium antagonist activity on guinea pig aorta strips with the negative inotropic activity, determined by using isolated guinea pig left atria, revealed that 3 displayed higher selectivity than the reference (+)-cis diltiazem. Thus, the pyrrolobenzothiazepine 3 might represent a new tool for characterizing the relationship between the PBR and cardiac function. Furthermore, we have also investigated the structural dependence of binding to PBR and L-type calcium channels, and this study allowed us to identify a new class of potent calcium channel blockers selective for cardiac over vascular tissue, with no affinity for PBR. A number of structure-activity relationship trends have been identified, and a possible explanation is advanced in order to account for the observed differences in selectivity. Three structural features, namely, (i) the saturation of the C(6)-C(7) double bond, with a consequent higher molecular flexibility, (ii) the presence of a substituent in the benzofused ring, and (iii) a basic side chain at C-10 of the pyrrolobenzothiazepine ring system, were found to be responsible for potent L-type calcium channel antagonism and clear-cut selectivity for cardiac over vascular tissue. Among the synthesized compounds the pyrrolobenzothiazepine 62 was found to be the most promising selective calcium channel blocker. Additionally, the molecular structure determination of the key intermediate 48 by X-ray diffraction, molecular modeling, and NMR analysis is reported. PMID- 8709128 TI - Similarities and differences in the structure-activity relationships of capsaicin and resiniferatoxin analogues. AB - Structure-activity relationships in analogues of the irritant natural product capsaicin have previously been rationalized by subdivision of the molecule into three structural regions (A,B, and C). The hypothesis that resiniferatoxin (RTX), which is a high-potency ligand for the same receptor and which has superficial structural similarities with capsaicin, could be analogously subdivided has been investigated. The effects of making parallel changes in the two structural series have been studied in a cellular functional assay which is predictive of analgesic activity. Parallel structural changes in the two series lead to markedly different consequences on biological activity; the 3- and 4-position aryl substituents (corresponding to the capsaicin 'A-region') which are strictly required for activity in capsaicin analogues are not important in RTX analogues. The homovanillyl C-20 ester group in RTX (corresponding to the capsaicin 'B region') is more potent than the corresponding amide, in contrast to the capsaicin analogues. Structural variations to the diterpene moiety suggest that the functionalized 5-membered diterpene ring of RTX is an important structural determinant for high potency. Modeling studies indicate that the 3D position of the alpha-hydroxy ketone moiety in the 5-membered ring is markedly different in the phorbol (inactive) analogues and RTX (active) series. This difference appears to be due to the influence of the strained ortho ester group in RTX, which acts as a local conformational constraint. The reduced activity of an analogue substituted in this region and the inactivity of a simplified analogue in which this unit is entirely removed support this conclusion. PMID- 8709129 TI - Dihydrobenzofuran analogues of hallucinogens. 3. Models of 4-substituted (2,5 dimethoxyphenyl)alkylamine derivatives with rigidified methoxy groups. AB - Tetrahydrobenzodifuran functionalities were employed as conformationally restricted bioisosteres of the aromatic methoxy groups in prototypical hallucinogenic phenylalkylamines 1 and 2. Thus, a series of 8-substituted 1 (2,3,6,7-tetrahydrobenzo[1,2-b:4,5-b']difuran-4-yl)-2-aminoal kanes (7a-e) were prepared and evaluated for activity in the two-lever drug discrimination paradigm in rats trained to discriminate saline from LSD tartrate (0.08 mg/kg) and for the ability to displace [3H]ketanserin from rat cortical homogenate 5-HT2A receptors and [3H]-8-OH-DPAT from rat hippocampal homogenate 5-HT1A receptors. In addition, 1-(8-bromo-2,3,6,7-tetrahydrobenzo[1,2-b:4,5-b']difuran-4-yl)-2-am inopropane (7b), which was found to be extremely potent in the rat in vivo assays, was evaluated for its ability to compete with [125I]DOI and [3H]ketanserin binding to cells expressing cloned human 5-HT2A, 5-HT2B, and 5-HT2C receptors. All of the dihydrofuranyl compounds having a hydrophobic substituent para to the alkylamine side chain had activities in both the in vitro and in vivo assays that equaled or surpassed the activity of the analogous conformationally flexible parent compounds. For example, 7b substituted for LSD in the drug discrimination assay with an ED50 of 61 nmol/kg and had Kj values in the nanomolar to subnanomolar range for the displacement of radioligand from rat and human 5-HT2 receptors, making it one of the most potent hallucinogen-like phenylalkylamine derivatives reported to date. The results suggest that the dihydrofuran rings in these new analogues effectively model the active binding conformations of the methoxy groups of the parent compounds 1 and 2. In addition, the results provide information about the topography and relative orientation of residues involved in agonist binding in the serotonin 5-HT2 receptors. PMID- 8709130 TI - 7-[3-(1-piperidinyl)propoxy]chromenones as potential atypical antipsychotics. AB - Compound 1 (1-benzyl-3-methyl-4-[4-(4-fluorophenyl)-4-oxobutyl]piperazine), a synthetic intermediate identified as a potential atypical antipsychotic, was selected as the starting point for pharmacological improvement. From 1, sequential structural variations were conducted in order to improve its potency and oral bioavailability. These variations included a series of piperazine, ethanediamine, and piperidine derivatives. The piperidine series afforded some orally potent compounds in the inhibition of apomorphine-induced climbing and hyperactivity in mice, which are regarded as behavioral models predictive of antipsychotic efficacy. Further optimization of these structures led to the highly potent 7-[3-(1-piperidinyl)propoxy]chromenones. Inhibition of stereotypies and induction of catalepsy in rats at doses substantially higher than required for inhibition of climbing suggest an atypical antipsychotic profile, which is assumed to predict a reduced induction of extrapyramidal side effects in humans. PMID- 8709131 TI - Synthesis and activity of a novel series of 3-biarylquinuclidine squalene synthase inhibitors. AB - Quinuclidines with a 3-biaryl substituent are a new class of potent, orally active squalene synthase (SQS) inhibitors. Variants around these rigid structures indicate key structural requirements for cationic SQS inhibitors. Thus the lower in vitro potency found for quinuclidines bearing 3-substituents, which did not overlay the biphenyl group of 3-(biphenyl-4-yl)-3-hydroxyquinuclidine (2) (IC50 = 16 nM, rat microsomal SQS), implied a directional requirement for the 3 substituent. Similarly, the lower potency of the 3-terphenyl analogue 6 (IC50 = 370 nM) indicated size constraints for this substituent. In compounds with a linking group between the quinuclidine and biphenyl ring, linking groups of lower lipophilicity were less well tolerated (e.g., 17, CH2CH2, IC50 = 5 nM vs 19, NHCO, IC50 = 1.2 microM). Replacement of the distal phenyl ring of 2 with a more polar pyridine heterocycle caused a reduction in in vitro potency. In general, good in vivo activity in the rat was restricted to 3-hydroxy analogues, with the 3-[4-(pyrid-4-yl)phenyl] derivative 39 (IC50 = 161 nM) showing the best inhibition (following oral dosing) of cholesterol biosynthesis from mevalonate (ED50 = 2.7 mg/kg). PMID- 8709132 TI - Interaction of 1,4-dihydropyridine and pyridine derivatives with adenosine receptors: selectivity for A3 receptors. AB - 1,4-Dihydropyridine and pyridine derivatives bound to three subtypes of adenosine receptors in the micromolar range. Affinity was determined in radioligand binding assays at rat brain A1 and A2A receptors using [3H]-(R)-PIA [[3H]-(R)-N6 (phenylisopropyl)adenosine] and [3H]CGS 21680 [[3H]-2-[[4-(2 carboxyethyl)phenyl]ethylamino]-5'-(N-ethylcarbamoyl++ +) adenosine], respectively. Affinity was determined at cloned human and rat A3 receptors using [125I]AB-MECA [N6-(4-amino-3-iodobenzyl)-5'-(N-methylcarbamoyl)adenosine]. Structure-activity analysis at adenosine receptors indicated that sterically bulky groups at the 4-, 5-, and 6-positions are tolerated. (R,S)-Nicardipine, 12, displayed Ki values of 19.6 and 63.8 microM at rat A1 and A2A receptors, respectively, and 3.25 microM at human A3 receptors. Similarly, (R)-niguldipine, 14, displayed Ki values of 41.3 and 1.90 microM at A1 and A3 receptors, respectively, and was inactive at A2A receptors. A preference for the R- vs the S enantiomer was observed for several dihydropyridines at adenosine receptors, in contrast with the selectivity at L-type Ca2+ channels. A 4-trans-beta-styryl derivative, 24, with a Ki value of 0.670 microM at A3 receptors, was 24-fold selective vs A1 receptors (Ki = 16.1 microM) and 74-fold vs A2A receptors (Ki = 49.3 microM). The affinity of 24 at L-type Ca2+ channels, measured in rat brain membranes using [3H]isradipine, indicated a Ki value of 0.694 microM, and the compound is thus nonselective between A3 receptors and L-type Ca2+ channels. Inclusion of a 6-phenyl group enhanced A3 receptor selectivity: Compound 28 (MRS1097; 3,5-diethyl 2-methyl-6-phenyl-4-(trans-2-phenylvinyl)-1,4(R,S)-dihydro pyridin e-3, 5-dicarboxylate) was 55-fold selective vs A1 receptors, 44-fold selective vs A2A receptors, and over 1000-fold selective vs L-type Ca2+ channels. In addition, compound 28 attenuated the A3 agonist-elicited inhibitory effect on adenylyl cyclase. Furthermore, whereas nicardipine, 12, displaced radioligand from the Na(+)-independent adenosine transporter with an apparent affinity of 5.36 +/- 1.51 microM, compound 28 displaced less than 10% of total binding at a concentration of 100 microM. Pyridine derivatives, when bearing a 4-alkyl but not a 4-phenyl group, maintained affinity for adenosine receptors. These findings indicate that the dihydropyridines may provide leads for the development of novel, selective A3 adenosine antagonists. PMID- 8709133 TI - Synthesis of the four isomers of 4-aminopyrrolidine-2,4-dicarboxylate: identification of a potent, highly selective, and systemically-active agonist for metabotropic glutamate receptors negatively coupled to adenylate cyclase. AB - The four isomers of 4-aminopyrrolidine-2,4-dicarboxylate (APDC) were prepared and evaluated for their effects at glutamate receptors in vitro. (2R,4R)-APDC (2a), an aza analog of the nonselective mGluR agonist (1S,3R)-1-aminocyclopentane-1,3 dicarboxylate (1S,3R)-ACPD, 1), was found to possess relatively high affinity for metabotropic glutamate receptors (mGluRs) (ACPD-sensitive [3H]glutamate binding IC50 = 6.49 +/- 1.21 microM) with no effects on radioligand binding to NMDA, AMPA, or kainate receptors up to 100 microM. None of the other APDC isomers showed significant mGluR binding affinity, indicating that this interaction is highly stereospecific. Both 1 and 2a were effective in decreasing forskolin stimulated cAMP formation in the adult rat cerebral cortex (EC50 = 8.17 +/- 2.21 microM for 1; EC50 = 14.51 +/- 5.54 microM for 2a); however, while 1 was also effective in stimulating basal tritiated inositol monophosphate production in the neonatal rat cerebral cortex (EC50 = 27.7 +/- 5.2 microM), 2a (up to 100 microM) was ineffective in stimulating phosphoinositide hydrolysis in this tissue preparation, further supporting our previous observations that 2a is a highly selective agonist for mGluRs negatively coupled to adenylate cyclase. Microelectrophoretic application of either 1 or 2a to intact rat spinal neurons produced an augmentation of AMPA-induced excitation (95 +/- 10% increase for 1, 52 +/- 6% increase for 2a). Intracerebral injection of 1 (400 nmol) produced characteristic limbic seizures in mice which are not mimicked by 2a (200-1600 nmol, ic). However, the limbic seizures induced by 1 were blocked by systemically administered 2a in a dose-dependent manner (EC50 = 271 mg/kg, ip). It is concluded that (2R,4R)-APDC (2a) is a highly selective, systemically-active agonist of mGluRs negatively coupled to adenylate cyclase and that selective activation of these receptors in vivo can result in anticonvulsant effects. PMID- 8709134 TI - Medetomidine analogs as alpha 2-adrenergic ligands. 2. Design, synthesis, and biological activity of conformationally restricted naphthalene derivatives of medetomidine. AB - A new series of naphthalene analogs of medetomidine have been prepared and evaluated for their alpha-adrenergic activities. The methylnaphthyl analog 5a showed significant selectivity for alpha 2-adrenoceptors and behaved as a partial alpha 1-agonist in rat aorta preparations. In contrast, the Z-ethylene analog 8c was alpha 1-selective and behaved as a potent alpha 1-antagonist. Two rigid analogs (6 and 7) exhibited large differences in binding affinities at alpha 1-VS alpha 2-receptors, indicating that the conformational flexibility of 5a is important for the fulfillment of the alpha-adrenergic activities. Molecular modeling studies began with conformational analysis of classical phenethylamines and medetomidine analogs. Superimposition of medetomidine conformations with those of phenethylamines provided a tentative explanation for the alpha 2 adrenergic activity of the new imidazoles. A common binding mode for phenethylamines and imidazoles with alpha 2-adrenoceptors is proposed. Knowledge of the biological properties of the 4-substituted imidazoles, integrated with the information derived from computer-assisted molecular modeling, has provided new insights for the structural and conformational requirements of this class as new adrenergic drugs. PMID- 8709135 TI - Identification of a 3-hydroxylated tacrine metabolite in rat and man: metabolic profiling implications and pharmacology. AB - Discrepancies in urinary metabolic profiles in rats administered tacrine (1) suggested the presence of an unidentified metabolite of 1. Chromatographic methods were developed that allowed isolation of a metabolite fraction containing both 1-hydroxytacrine (2) and an unknown metabolite from rat urine. Mass spectral analysis indicated this metabolite to be a monohydroxylated derivative, which upon two dimensional COSY NMR analysis could be assigned as 3-hydroxytacrine (4). This structural assignment was confirmed by independent synthesis of 4. Compound 4 was also identified as a human urinary metabolite of 1. Biologically, 4 was found to have in vitro human red blood cell acetylcholinesterase inhibitory activity similar to that of 2 and 4-hydroxytacrine (5) and approximately 8-fold less than that of 1. These results underscore the need to conduct rigorous structural identification studies, especially in cases where isomeric metabolites are possible, in assessing the accuracy of chromatographic profiling techniques. PMID- 8709136 TI - Synthesis and pharmacological activity of triazole derivatives inhibiting eosinophilia. AB - In order to develop novel antiasthmatic agents based on a new mechanism of action, a series of 3-substituted 5-amino-1-[(methylamino)(thiocarbonyl)]-1H 1,2,4-triazole derivatives were synthesized and evaluated in a model in which eosinophilia was induced in the airway through intravenous (iv) injection of Sephadex particles on days 0, 2, and 5. After screening of several hundred derivatives, we finally identified the highly potent eosinophilia inhibitor 5 amino-3-(4-chlorophenyl)-1-[(methylamino)(thiocarbonyl)]-1H-tria zole (23c, GCC AP0341), which had ID50 values of 0.3 and 0.07 mg/kg when administered orally (os) and intraperitoneally (ip), respectively. This compound showed complete inhibition of the hypersensitivity induced by ascaris inhalation at an ip dose of 1 mg/kg as well as low toxicity, with an LD50 value of > 2.0 g/kg in mice. Extensive study of its mechanism of action revealed that 23c inhibited eosinophil survival induced by interleukin-5 (IL-5), but had little or no effect on leukotriene D4 (LTD4) or platelet-activating factor (PAF)-induced responses. Taken together, these results suggest 23c as a novel candidate for the treatment of chronic asthma. Further studies are now underway. PMID- 8709137 TI - 3-[2-(N-phenylacetamide)]-1,5-benzodiazepines: orally active, binding selective CCK-A agonists. AB - A series of modifications were made to the C-3 substituent of the 1,5 benzodiazepine CCK-A agonist 1. Replacement of the inner urea NH and addition of a methyl group to generate a C-3 quaternary carbon resulted in acetamide 6, which showed CCK-A receptor binding selectivity and sub-micromolar agonist activity in vitro. Benzodiazepine 6 was active in an in vivo mouse gallbladder emptying assay and represents a novel orally active, binding selective CCK-A agonist. PMID- 8709138 TI - Heart failure: epidemiology, economics and treatment. PMID- 8709139 TI - Peritonsillar abscess following tonsillectomy. AB - Peritonsillar abscess is an unusual complication following tonsillectomy. A series of five such patients is herein reported. Proper treatment begins with the initial tonsillectomy ensuring complete removal of the tonsils. Should a late peritonsillar abscess following tonsillectomy be found, the patient may be treated with needle aspiration of the affected area plus antibiotics. Removal of the remaining tonsillar tissue and surgical capsule of the tonsil is also advocated. PMID- 8709140 TI - If it is to be--it is up to me. PMID- 8709141 TI - "Fischerisms". PMID- 8709142 TI - Case records of Department of Medicine University of Mississippi Medical Center Diagnosis. PMID- 8709143 TI - Structural aspects of the p53 protein in relation to gene evolution: a second look. AB - Several years ago, a comparison of the amino acid sequences of p53 proteins from a variety of species enabled us to reveal structural features of this protein, giving clues to its function. Since then, numerous studies on the biochemical, immunological and biological functions of p53 as well as on its structure (including crystallography data) have provided considerable insight into the multifunctional aspects of p53. The purpose of this review is to present the most recent data concerning the various structural features of the p53 protein with special emphasis on its flexibility, which plays a key role in regulation of its biological activity. PMID- 8709144 TI - The structure of recombinant human annexin VI in crystals and membrane-bound. AB - The crystal structure of calcium-free recombinant human annexin VI was solved at a resolution of 3.2 A by using the annexin I model for Patterson search and refined to an R-factor of 19.0%. The molecule consists of two similar halves closely resembling annexin I connected by an alpha-helical segment and arranged perpendicular to each other. The calcium and membrane binding sites assigned by structural homology are therefore not located in the same plane. Analysis of the membrane-bound form of annexin VI by electron microscopy shows the two halves of the molecule coplanar with the membrane, but oriented differently to the crystal structure and suggesting a flexible arrangement. Ion channel activity has been found for annexin VI and the half molecules by electrophysiological experiments. PMID- 8709145 TI - Helmholtz free energy of peptide hydrogen bonds in proteins. AB - We estimate the Helmholtz free energy of peptide hydrogen bonds in native protein structures as a function of spatial separation between donor and acceptor atoms. The resulting potential function has a deep narrow well at H-bond contact but bond formation is hindered by a barrier and the net change in free energy is close to zero. The barrier provides a molecular lock mechanism acting as a kinetic trap. Once formed, H-bonds keep protein chains in a precise orientation. However, bond formation requires energy input and opposes protein folding. In contrast, the free energy functions of most side-chain interactions have no energy barriers. They lack spatial precision but free energy differences of contact formation are substantial. These interactions drive folding and stabilize structures but precision is mediated and maintained by H-bonds. PMID- 8709146 TI - Co-variation of tRNA abundance and codon usage in Escherichia coli at different growth rates. AB - We have used two-dimensional polyacrylamide gel electrophoresis to fractionate tRNAs from Escherichia coli. A sufficiently high degree of resolution was obtained for 44 out of 46 tRNA species in E. coli to be resolved into individual electrophoretic components. These isolated components were identified by hybridization to tRNA-specific oligonucleotide probes. Systematic measurements of the abundance of each individual tRNA isoacceptor in E. coli, grown at rates varying from 0.4 to 2.5 doublings per hour, were made with the aid of this electrophoretic protocol. We find that there is a biased distribution of the tRNA abundance at all growth rates, and that this can be roughly correlated with the values of codon frequencies in the mRNA pools calculated for bacteria growing at different rates. The tRNA species cognate to abundant codons increase in concentration as the growth rate increases but not as dramatically as might be anticipated. The levels of most of the tRNA isoacceptors cognate to less abundant codons remain unchanged with increasing growth rates. The result of these changes in tRNA abundance is that the relative increase in the amounts of major tRNA species in the bacteria growing at the fastest growth rates is more modest than previous estimates from this laboratory suggested. Furthermore, a systematic error in previous estimates of ribosomal RNA content of the bacteria has been detected. This will account for the quantitative discrepancies between the previous and the present data for tRNA abundance. PMID- 8709147 TI - Catalytic specificity of phosphotyrosine kinases Blk, Lyn, c-Src and Syk as assessed by phage display. AB - Protein tyrosine kinases (PTKs) are implicated in cell proliferation, differentiation, and receptor-mediated signalling events. Recruitment of intracellular PTKs into the signalling complex, often localized at the inner surface of the cell membrane, involves SH2 and SH3 domains attached to the catalytic kinase domain. While the interaction of SH2 and SH3 domains with their target sequences is well documented in a number of cases, the contribution of the catalytic domain itself in conferring specificity to a given signal cascade is not fully understood. We addressed this question and employed the phage display technique to assess the substrate requirements for the highly related Src-like PTKs c-Src, Blk, Lyn and the distantly related Syk. A diverse peptide library on phage was established, and after multiple rounds of phosphorylation and selection of phage displaying phosphotyrosine-containing peptides, canonical substrate sequences for each of the PTKs were enriched. The PTKs Blk and Lyn implicated in B cell signalling were found to prefer peptide substrates of the structure I/L-Y D/E-X-L which resemble critical features of the ITAM motifs found in, e.g. the intracellular components Ig-alpha and Ig-beta of the beta cell receptor. All Src like PTKs had a requirement for isoleucine or leucine in the position -1 with respect to the phosphorylated tyrosine residue in position 0. While Blk and Lyn had a strong preference for a negatively charged amino acid in position +1, c-Src preferred tryptophan or glycine in this position. Syk, not belonging to the Src like PTK family, revealed a distinct substrate requirement for aspartic acid in position -1 and glutamic acid in position +1. In general, all PTKs we have tested had a strong preference for a particular amino acid in the positions -1 and +1 adjacent to the tyrosine residue. PMID- 8709148 TI - T4 endonuclease VII selects and alters the structure of the four-way DNA junction; binding of a resolution-defective mutant enzyme. AB - Bacteriophage T4 endonuclease VII is a nuclease that is selective for four-way DNA junctions and related branched DNA species. Using site-directed mutagenesis we have isolated a mutant protein (E86A) that is inactive in the cleavage of DNA junctions while retaining full selectivity of binding. Using endonuclease VII E86A we have shown: (1) The protein binds as a dimer to DNA junctions, with rapid exchange of subunits in free solution. (2) Binding to junctions is highly selective for the structure of DNA junctions; the complex is not displaced by a 1000-fold excess of duplex competitor DNA. (3) On binding endonuclease VII E86A to junctions, the configuration of the helical arms is significantly altered to a structure that is independent of the presence or absence of metal ions. We suggest a model for the structure of the junction in the protein complex. (4) The protein can bind to the junction in two stereochemically equivalent ways, depending upon the sequence of the junction. T4 endonuclease VII is a junction selective enzyme that both recognises and manipulates the structure of its substrate. PMID- 8709149 TI - Cooperative and anticooperative effects in binding of the first and second plasmid Osym operators to a LacI tetramer: evidence for contributions of non operator DNA binding by wrapping and looping. AB - The interaction of lac operator DNA with lac repressor (LacI) is a classic example of a genetic regulatory switch. To dissect the role of stoichiometry, subunit association, and effects of DNA length in positioning this switch, we have determined binding isotherms for the interaction of LacI with a high affinity (Osym) operator on linearized plasmid (2500 bp) DNA over a wide range of macromolecular concentrations (10(-14) to 10(-8) M). Binding data were analyzed using a thermodynamic model involving four equilibria: dissociation of tetramers (T) into dimers (D), and binding of operator-containing plasmid DNA (O) to dimers and tetramers to form three distinct complexes, DO, TO, and TO2. Over the range of concentrations of repressor, operator, and salt (0.075 M K+ to 0.40 M K+) investigated, we find no evidence for any significant thermodynamic effect of LacI dimers. Instead, all isotherms can be interpreted in terms of just two equilibria, involving only T and the TO and TO2 complexes. As a reference binding equilibrium, which we propose must approximate the DO binding interaction, we compare the plasmid Osym results with our extensive studies of the binding of a 40 bp Osym DNA fragment to LacI. On this basis, we obtain a lower bound on the LacI dimer-tetramer equilibrium constant and values of the equilibrium constants for formation of TO and TO2 complexes. At a salt concentration of 0.40 M, the Osym plasmid binding data are consistent with a model with two independent and identical binding sites for operator per LacI tetramer, in which the binding to a site on the tetramer is only slightly more favorable than the reference binding interaction. Increasingly large deviations from the independent-site model are observed as the salt concentration is reduced; binding of a second operator to from TO2 becomes strongly disfavored relative to formation of TO at low salt concentrations (0.075 to 0.125 M). In addition, binding of both the first and second plasmid operator DNA molecules to the tetramer becomes increasingly more favorable than the reference binding interaction as [K+] is reduced from 0.40 M to 0.125 M. At 0.075 M K+, however, the strength of binding of the second plasmid operator DNA to the LacI tetramer is dramatically reduced; this interaction is much less favorable than binding the first plasmid operator DNA, and becomes much less favorable than the reference binding interaction. We propose that these differences arise from changes in the nature of the TO and TO2 complexes with decreasing salt concentration. At low salt concentration, we suggest the hypothesis that flanking non-operator sequences bind non-specifically (coulombically) by local wrapping, and that distant regions of non-operator DNA occupy the second operator-binding site by looping. We propose that wrapping stabilizes both 1:1 and 2:1 complexes at low salt concentration, and that looping stabilizes the 1:1 complex but competitively destabilizes the 2:1 TO2 complex at low salt concentration. These effects must play a role in adjusting the stability and structure of the LacI-lac operator repression complex as the cytoplasmic [K+] varies in response to changes in extracellular osmolarity. PMID- 8709150 TI - Effect of bending strain on the torsion elastic constant of DNA. AB - The torsion constants of both circular and linear forms of the same 181 bp DNA were investigated by time-resolved fluorescence polarization anisotropy (FPA) of intercalated ethidium. The ratio of intrinsic ethidium binding constants of the circular and linear species was determined from the relative fluorescence intensities of intercalated and non-intercalated dye in each case. Possible changes in secondary structure were also probed by circular dichroism (CD) spectroscopy. Upon circularization, the torsion constant increased by a factor of 1.42, the intrinsic binding constant for ethidium increased by about fourfold, and the CD spectrum underwent a significant change. These effects are attributed to an altered secondary structure induced by the bending strain. Quantitative agreement between torsion constants obtained from the present FPA studies and previous topoisomer distribution measurements on circular DNAs containing 205 to 217 bp removes a long-standing apparent discrepancy between those two methods. After storage at 4 degrees C for eight months, the torsion constant of the circular DNA increased by about 1.25-fold, whereas that of the linear DNA remained unchanged. For these aged circles, both the torsion constant and intrinsic binding constant ratio lie close to the corresponding values obtained previously for a 247 bp DNA by analyzing topoisomer distributions created in the presence of various amounts of ethidium. The available evidence strongly implies that torsion constants measured for small circular DNAs with less than 250 bp are specific to the altered secondary structure(s) therein, and are not applicable to linear and much larger circular DNAs with lower mean bending strains. PMID- 8709151 TI - Identification of putative active-site residues in the DNase domain of colicin E9 by random mutagenesis. AB - We have used random mutagenesis to identify putative active-site residues in the C-terminal cytotoxic endonuclease domain of the bacterial toxin colicin E9. Six single-site mutations in the DNase domain were isolated which destroyed the toxic action of the colicin. DNA sequencing identified the mutations as Gly460Asp, Arg544Gly, Glu548Gly, Thr571Ile, His575Tyr and His579Tyr. All six wild-type residues are highly conserved in the DNase domains of both the E group colicins and the closely related pyocins. Site-directed mutagenesis was then used to substitute the wild-type amino acid residue at each of these positions for an alanine residue in order to distinguish important from unimportant sites. Two of the six alanine-mutant colicins (Gly460Ala and His579Ala) exhibited significant in vivo activity, unlike the original mutation of these residues, and were therefore not characterised further. The Thr571Ala mutant colicin, although not inactive, was significantly less active than the control. The other three alanine mutants (Arg544Ala, Glu548Ala and His575Ala remained completely inactive in the in vivo tests. Each 15 kDa alanine-mutant DNase domain was overexpressed and purified using a tandem-expression strategy which relies on the enzyme being able to bind to the natural inhibitor, Im9. Tryptophan emission spectra of the alanine mutants showed significant alterations in the emission maxima of all but the His575Ala mutant, suggesting changes in the tertiary structure of these mutant proteins. Activity measurements, using the spectrophotometric Kunitz assay, indicated that the Thr571Ala mutant was partially active as an endonuclease but the remaining alanine mutants were all completely inactive. All four mutant proteins, however, retained their ability to bind DNA in a gel shift assay, suggesting the mutations affect catalytic rather than substrate-binding residues. Searching the sequence databases for possible homology to other DNA-binding proteins revealed a significant match between residues 464 to 487 of the E9 DNase domain and helix IV of the POU domain of eukaryotic transcription factors. PMID- 8709152 TI - Three-dimensional structure of ncd-decorated microtubules obtained by a back projection method. AB - We have used cryo-electron microscopy and image analysis to obtain the three dimensional (3D) structure of 11, 12, 14 and 15 protofilament microtubules decorated with the motor domain of ncd. To obtain the 3D maps, we developed a back-projection method that does not require a helical arrangement of the tubulin heterodimers. This method allows the calculation of 3D maps even when lattice discontinuities (seams) are present. The maps show that the microtubules we studied conform to a B-type lattice with one or more seams. In the presence of 5' adenylim-idodiphosphate (AMP-PNP), the motor domain of ncd binds to the microtubule protofilament crest interacting with only one protofilament. Viewing the structures along the microtubule axis shows that the ncd motor domain and the tubulin are titled in opposite directions. We determined that a clockwise tilt of the tubulin subunits corresponds to a view from the minus end towards the plus end of the microtubule. PMID- 8709153 TI - Cooperativity in F-actin: binding of gelsolin at the barbed end affects structure and dynamics of the whole filament. AB - We have studied the effect of gelsolin, a Ca-dependent actin-binding protein, on the microsecond rotational dynamics of actin filaments, using time-resolved phosphorescence (TPA) and absorption anisotropy (TAA) of erythrosin iodoacetamide attached to Cys374 on actin. Polymerization of actin in the presence of gelsolin resulted in substantial increases in the rate and amplitude of anisotropy decay, indicating increased rotational motion. Analysis indicates that the effect of gelsolin cannot be explained by increased rates of overall (rigid-body) rotations of shortened filaments, but reflects changes in intra-filament structure and dynamics. We conclude that gelsolin induces (1) a 10 degrees change in the orientation of the absorption dipole of the probe relative to the actin filament, indicating a conformational change in actin, and (2) a threefold decrease in torsional rigidity of the filament. This result, which is consistent with complementary electron microscopic observations on the same preparations, directly demonstrates long-range cooperativity in F-actin, where a conformational change induced by the binding of a single gelsolin molecule to the barbed end is propagated along inter-monomer bonds throughout the actin filament. PMID- 8709154 TI - Stoichiometry and domainal organization of the long tail-fiber of bacteriophage T4: a hinged viral adhesin. AB - The long-tail fibers (LTFs) form part of bacteriophage T4's apparatus for host cell recognition and infection, being responsible for its initial attachment to susceptible bacteria. The LTF has two parts, each approximately 70 to 75 nm long; gp34 (140 kDa) forms the proximal half-fiber, while the distal half-fiber is composed of gp37 (109 kDa), gp36(23 kDa) and gp35 (30 kDa). LTFs have long been thought to be dimers of gp34, gp37 and gp36, with one copy of gp35. We have used mass mapping by scanning transmission electron microscopy (STEM), quantitative SDS-PAGE, and computational sequence analysis to study the structures of purified LTFs and half-fibers of both kinds. These data establish that the LTF is, in fact, trimeric, with a stoichiometry of gp34: gp37: gp36: gp35 = 3:3:3:1. Averaged images of stained and unstained molecules resolve the LTF into a linear stack of 17 domains. At the proximal end is a globular domain of approximately 145 kDa that becomes incorporated into the baseplate. It is followed by a rod like shaft (33 x 4 mm; 151 kDa) which correlates with a cluster of seven quasi repeats, each 34 to 39 residues long. The proximal half-fiber terminates in three globular domains. The distal half-fiber consists of ten globular domains of variable size and spacing, preceding a needle-like end domain (15 x 2.5 nm; 31 kDa). The LTF is rigid apart from hinges between the two most proximal domains, and between the proximal and distal half-fibers. The latter hinge occurs at a site of local non-equivalence (the "kneecap") at which density, correlated with the presence of gp35, bulges asymmetrically out on one side. Several observations indicate that gp34 participates in the sharing of conserved structural modules among coliphage tail-fiber genes to which gp37 was previously noted to subscribe. Two adjacent globular domains in the proximal half-fiber match a pair of domains in the distal half-fiber, and the rod domain in the proximal half-fiber resembles a similar domain in the T4 short tail-fiber (gp12). Finally, possible structures are considered; combining our data with earlier observations, the most likely conformation for most of the LTF is a three-stranded beta-helix. PMID- 8709155 TI - [Histopathological and electron microscopic changes in mice treated with puffer fish toxin]. AB - The histopathological examination of a male ddY mouse given an intraperitoneal injection of the crude extract containing 169 micrograms/ml of tetrodotoxin (TTX) prepared from the ovary of Takifugu porphyreus was carried out. Two crude extract solutions (0.42 and 0.21 microgram/ml) were prepared; one kills an animal in 10 min (group 1), and other kills it in 40 min (group 2) after the injection. As a result, a remarkable inflation of the gall bladder was observed macroscopically in both groups. By light microscopic observed, both groups showed the congestion in the brain, heart, liver, gall bladder, lung and kidneys, while nerve cells were characterized by the vacuolation and disappearance of Nissl's body only in group 2, probably because of the mitochondrial degeneration. Electron microscopically, a part of cristae of mitochondria and ribosome of endoplasmic reticulum disappeared or vacuolated in the nerve cells of group 2. This was also observed in a part of pre and postsynaptic mitochondria of synapse. However, little significant changes were observed in group 1 and in other tissues of group 2. These results show that a cytopathological change of mice by TTX is mainly detected in nerve tissues when a small amount of TTX is administered to the animals. PMID- 8709156 TI - [A 13-week oral toxicity study of prulifloxacin (NM441) in dogs followed by a 5 week recovery test]. AB - A repeated dose toxicity study of prulifloxacin, a new antibacterial agent, was conducted in beagle dogs. Male and female dogs were given the test material orally for 13 weeks at doses of 0 (control), 20, 100 and 500 mg/kg. After discontinuation of the treatment, a 5-week recovery test was also conducted. Vomiting, salivation and decreased body weight gain or reduced body weight were seen in the 100 and 500 mg/kg groups. In the 500 mg/kg group, tremor, paresis of posterior limb associated with prone or sitting position and decreased food consumption were also observed. There were no treatment-related effects on survival and water consumption. Ophthalmoscopic, electrocardiographic and hematologic examinations, and urinalysis failed to show any abnormalities attributable to the treatment. Blood chemical examination showed increased GPT and decreased beta- and gamma-globulins in the 100 and 500 mg/kg groups, and increased GOT in the 500 mg/kg group. In pathological examination, cavitations and erosions were seen in the humeral and femoral articular cartilages in the 100 and 500 mg/kg groups. The above-mentioned changes were satisfactorily reversible except for erosions in the humeral and femoral articular cartilages in the 100 and 500 mg/kg groups. No toxicological findings were seen in the 20 mg/kg group. The results show that the NOAEL of prulifloxacin is 20 mg/kg for 13-week repeated dose toxicity in dogs. PMID- 8709157 TI - [A 4-week intravenous toxicity study of the active metabolite (NM394) of prulifloxacin (NM441) in rats followed by a 4-week recovery test]. AB - A repeated dose toxicity study of ( +/- )-6- fluoro-1-methyl-4-oxo-7-(1 piperazinyl)-4H- [1,3]thiazeto[3,2-a]quinoline-3-carboxylic acid (NM394), the active metabolite of a new antibacterial agent, prulifloxacin, was conducted in Sprague-Dawley rats. Male and female rats were given the test material intravenously for 4 weeks at doses of 0 (control), 3, 10 and 30 mg/kg. After discontinuation of the treatment, a 4-week recovery test was also conducted. There were no treatment-related effects on survival, clinical signs, body weight and food consumption. Ophthalmoscopic and hematologic examinations failed to show any abnormalities related to the treatment. Increased water consumption was observed in the 10 and 30 mg/kg groups. In these dose groups, increased urine volume and lowered urine specific gravity, and crystalline substance and small epithelial cells in urinary sediments were seen. Cloudy urine was also seen in the 30 mg/kg group. Blood chemical examination showed decreased gamma-globulin in the 10 and 30 mg/kg groups and increased BUN and creatinine in the 30 mg/kg group. Pathological changes caused by the treatment were as follows. In kidney, tubular nephrosis with crystalline substance was observed in the 10 and 30 mg/kg groups and its organ weight was increased in the 30 mg/kg group. Cecal weight was increased in the 30 mg/kg group. The above-mentioned changes were reversible except for decreased gamma-globulin. Plasma levels and urinary concentrations of the test material were increased in all dose groups with dose-related manner, whereby no sex differences were observed. No effects caused by the repeated dosing were seen in the plasma concentrations. Toxicological findings were not observed in the 3 mg/kg group. The results show that the NOAEL of NM394 is 3 mg/kg for 4-week repeated dose toxicity in rats. PMID- 8709158 TI - [Single and 4-week oral toxicity studies of prulifloxacin (NM441) in aged dogs]. AB - Single-dose and repeated dose toxicity studies of prulifloxacin, a new antibacterial agent, were conducted in aged beagle dogs. I. A single-dose toxicity study Prulifloxacin was administered orally to aged female dogs at a single dose of 2500 and 5000 mg/kg. No death occurred in any group. Vomiting was observed in one of two animals at 2500 mg/kg and in both animals at 5000 mg/kg 3 4 hr after dosing. At 5000 mg/kg, vomiting was observed in both animals after feeding on the day after dosing. One animal also showed soft stool. Thereafter, no abnormalities were observed in any animal. No test article related changes were noted in food consumption, water consumption, body weight or pathological examination in any group. The results show that the lethal dose of prulifloxacin is judged to be greater than 5000 mg/kg in aged female dogs. II. A repeated dose toxicity study Aged male and female dogs were given the test article orally for 4 weeks at doses of 0 (control), 20, 100 and 500 mg/kg. No death occurred in any group. At 500 mg/kg, vomiting was observed every day or intermittently throughout the dosing period and salivation was observed almost every day from day 6 to the end of the dosing onward. Decreases or lack of food and water consumption, and decrease of body weight were noted at 500 mg/kg. At 100 mg/kg, slight decreases in food consumption and body weight were noted in the females. No abnormalities were noted in ophthalmoscopic or electrocardio-graphic examination. In urinalysis, decreases in Na+, K+ and Cl- concentrations and the total excretion amount were noted mostly at 500 mg/kg. A low specific gravity was noted in males at 500 mg/kg. In hematology and serum biochemistry, high GPT, BUN and creatinine, and decreases in WBC were noted in both sexes at 500 mg/kg. A high GOT was noted in males, and low Cl- in females at 500 mg/kg. At 100 mg/kg, a high GPT was noted. Rough surface in the kidney and chronic interstitial nephritis (basophilic change of tubule, atrophy of tubule, thickening of tubular basement membrane, interstitial mononuclear cell infiltration, interstitial focal fibrosis) were increased at 500 mg/kg. No toxicological findings were seen in the 20 mg/kg group. The results show that the NOAEL of prulifloxacin is 20 mg/kg for 4-week repeated dose toxicity in aged dogs. PMID- 8709159 TI - [A procedure for recording electroretinogram (ERG) and effect of sodium iodate on ERG in mice]. AB - A procedure for recording the electroretinogram (ERG) in mice with a coiled stainless steel-type electrode was developed in order to examine retinal toxicity. Mice received a single i.v. of sodium iodate (SI), a retinotoxic compound, via the tail vein at a dose of 12.5, 25, or 50 mg/kg, and the ERG was recorded periodically for 28 days after dosing. In addition, the retina was examined histopathologically on day 30 after dosing. 1. The mice were anesthetized with mixed anesthetics of urethane, xylazine and ketamine after 30 to 60 min of dark-adaptation. Sixteen responses to repetitive 1.2 J light stimuli at a frequency of 0.2 or 0.1 Hz were averaged by a microcomputer. Body temperature of the mice was kept constant at 37 to 38 degrees C using a thermostatically controlled heating mat. Under these conditions, stable ERG a wave, b-wave, oscillatory potentials and c-wave could be recorded for 28 days. 2. SI at doses of 25 mg/kg or more caused depression of the amplitudes of the oscillatory potentials, and enhancement of the a- and b-wave amplitudes, while the c-wave was already extinguished on day 1 after dosing. Following these changes, the amplitudes of the a- and b-wave decreased from day 3 or 7 after dosing. These changes did not recover until day 28 after dosing. 3. Upon histopathologic examination of the retina, folding of the outer nuclear layer, disarrangement of the rods and cones, decrease of the visual cells and swelling and decrease of the pigment epithelial cells were observed with SI at 25 mg/kg or more. 4. Using this recording technique, it was confirmed that a stable ERG was recorded repeatedly for 28 days in mice, and the effects of SI on the ERG could be detected. Histopathologic findings in the retina revealed the abnormal portions were correlated well with the changes in the ERG. These results indicate that the ERG recording procedure developed in this study is useful for evaluating retinal toxicity in mice. PMID- 8709160 TI - [Reproductive and developmental toxicity studies of prulifloxacin (NM441)(1)--A fertility study in rats by oral administration]. AB - A study of fertility and fetal development of prulifloxacin, a new antibacterial agent, was conducted in Sprague-Dawley rats. Male rats were given the drug orally from 63 days before mating to the end of the mating period. Female rats were given from 14 days before mating to day 7 of pregnancy. The dose levels for both males and females were 0 (control), 30, 170 and 1000 mg/kg. The females were sacrificed on day 20 of pregnancy for examination of their fetuses. In the 170 mg/kg group, water consumption increased in the female rats, and food consumption decreased and the cecum increased in weight and was enlarged in the males and the females. In the 1000 mg/kg group, body weight gain was suppressed in the males, and food consumption decreased, water consumption increased and cecum increased in weight and enlarged in the males and the females. Moreover, this dose caused white spots and rough surface of the kidney in the males. Prulifloxacin had no adverse effects on reproductive function of the parent animals, and also on development of the fetuses. These results show that the NOAEL of prulifloxacin are 30 mg/kg for general toxicity in parent animals, 1000 mg/kg for reproductive function of parent animals and for development of fetuses. PMID- 8709161 TI - [Reproductive and developmental toxicity studies of prulifloxacin (NM441)(2)--A teratogenicity study in rats by oral administration]. AB - A study of the effect of prulifloxacin, a new antibacterial agent, during the period of organogenesis was conducted in Sprague-Dawley rats. Female rats were given prulifloxacin orally at dose levels of 0 (control), 30, 300, and 3000 mg/kg from day 7 to day 17 of pregnancy. Twenty-four or twenty-five female rats per dose level were sacrificed on day 20 of pregnancy for examination of their fetuses, and the pregnant rats (12-15 per dose levels) were allowed to deliver naturally for postnatal examination of their offspring. In the 300 or 3000 mg/kg groups, the food consumption decreased and water consumption increased in the dams. In the 3000 mg/kg group, the body weight gain was suppressed in the dams. In the 300 and 3000 mg/kg groups, the weights of cecum increased and the enlargement of cecum was observed. In the 3000 mg/kg group, white spots and rough surface of the kidney and renal tubular nephrosis with crystalline substance were observed and the weight of the kidney increased in the dams, and the body weight decreased and retarded ossifications in the fetuses. Prulifloxacin had no effect on the number of corpora lutea and implantations, or on fetal mortality, sex ratio, or external and visceral development of the fetuses. Prulifloxacin also did not affect delivery and the number of live newborns and birth index, or have any adverse effects on the postnatal development of the first generation offspring (F1) such as differentiation, functional development, emotionality, motor ability, learning ability or reproductive performance. Prulifloxacin also had no adverse effects of the second generation offspring (F2). These results show that the NOAEL of prulifloxacin are 30 mg/kg for general toxicity in mother animals, 3000 mg/kg for pregnancy and delivery of mother animals and 300 mg/kg for development of their offspring. PMID- 8709162 TI - [Reproductive and developmental toxicity studies of prulifloxacin (NM441)(3)--A teratogenicity study in rabbits by oral administration]. AB - A study of the effect of prulifloxacin, a new antibacterial agent, during the period of organogenesis was conducted in New Zealand White rabbits. Female rabbits were given prulifloxacin orally at dose levels of 0 (control), 10, 30 and 100 mg/kg from day 6 to 18 of pregnancy. Female rabbits were sacrificed on day 29 of pregnancy for examination of their fetuses. In the 30 mg/kg group, food and water consumption decreased. In the 100 mg/kg group, soft stool was observed and body weight gain, food consumption and water consumption decreased. Premature delivery (2/16) occurred and enlargement of cecum and increased weight of cecum were observed. The number of fetal death increased in the 100 mg/kg group. However, prulifloxacin had no effects on the number of corpora lutea, implantations and live fetuses, and on body weight, placental weight, sex ratio, and external, visceral and skeletal development of live fetuses. These results show that the NOAEL of prulifloxacin are 10 mg/kg for general toxicity in mother animals, 30 mg/kg for pregnancy of mother animals and for development of fetuses. PMID- 8709163 TI - [Reproductive and developmental toxicity studies of prulifloxacin (NM441)(4)--A perinatal and postnatal study in rats by oral administration]. AB - A peri- and postnatal study of prulifloxacin, a new antibacterial agent, was conducted in Sprague-Dawley rats. Female rats were given prulifloxacin orally at dose levels of 0 (control), 30, 100 and 300 mg/kg from day 17 of pregnancy to day 21 after delivery. All pregnant rats were allowed to deliver naturally for postnatal examination of their offspring. Food consumption in the period of pregnancy decreased in the dams given 100 and 300 mg/kg, and food and water consumption during lactational period increased in the dams given 300 mg/kg. The weight of the cecum of the dams increased in 100 and 300 mg/kg groups, and these doses caused enlargement of the cecum. Prulifloxacin had no effects on general sign, delivery, nursing, body weight gain, and kidney. Prulifloxacin also did not affect the number of live newborns and birth index, and have any adverse effects on the postnatal development of the first generation offspring (F1) such as differentiation, functional development, emotionality, motor ability, learning ability and reproductive performance. Prulifloxacin also had no adverse effects on the second generation offspring (F2). These results show that the NOAEL of prulifloxacin are 30 mg/kg for general toxicity in mother animals, 300 mg/kg for pregnancy and delivery of mother animals and development of their offspring. PMID- 8709164 TI - [Antigenicity studies of prulifloxacin (NM441)]. AB - Antigenicity of prulifloxacin, a new antibacterial agent, and that of its active metabolite (NM394) were investigated using guinea pigs and mice in this study. In the study with guinea pigs, the animals were immunized with prulifloxacin by oral administration, the predetermined clinical route, and with prulifloxacin alone or emulsified with Freund's complete adjuvant (FCA) by s.c. When active systemic anaphylaxis (ASA) test and passive cutaneous anaphylaxis (PCA) test of these animals were conducted by eliciting with NM394 alone or protein conjugate (NM394 GPSA) of NM394 and guinea pig serum albumin (GPSA), no ASA and PCA reactions were observed. When guinea pigs were immunized subcutaneously with protein conjugate (NM394-BSA) of NM394 and bovine serum albumin (BSA) together with FCA, ASA and PCA reactions were positive by eliciting with NM394-GPSA or NM394-BSA, but were negative by eliciting with NM394 alone. In the study with mice, the animals were immunized orally with prulifloxacin and intraperitoneally with prulifloxacin alone or suspended with aluminum hydroxide gel (alum). When rat PCA test of sera from these mice was conducted by eliciting with NM394 or NM394-GPSA, no positive PCA reaction was observed. When mice were immunized intraperiotneally with NM394 BSA together with alum, PCA reactions were positive by eliciting with NM394-GPSA or NM394-BSA, but were negative by eliciting with NM394 alone. These results show that prulifloxacin has no immunogenicity to guinea pigs and mice, and NM394 as its active form has no eliciting potential to anti-NM394-BSA antibody. PMID- 8709165 TI - [Mutagenicity studies of prulifloxacin (NM441) and the active metabolite (NM394)]. AB - The mutagenicity of prulifloxacin, a new antibacterial agent, was investigated by the reverse mutation test in bacteria, the chromosomal aberration test in cultured cells, and the micronucleus test in mice. In addition, NM394, an active metabolite of prulifloxacin, was examined for mutagenicity in the chromosomal aberration test in cultured cells. The reverse mutation test was performed at dose range of 0.0078-0.25 micrograms/plate using Salmonella typhimurium strains (TA100, TA1535, TA98, and TA1537), and Escherichia coli (WP2uvrA). Prulifloxacin did not increase revertant colonies significantly in any of the test strains with or without metabolic activation system (S9 mix). The chromosomal aberration tests were carried out in cultured Chinese hamster lung cells (CHL/IU). Prulifloxacin increased aberrant cells without S9 mix, and NM394 also induced chromosomal aberrations. In human lymphocytes, no significant increases of the frequencies of cells with chromosomal aberrations were observed at dose range of 5-320 micrograms/ml with or without S9 mix. The micronucleus test was conducted at doses of 625-5000 mg/kg in the bone marrow cells of Slc : ddY male mice. There were no significant increases in the frequencies of micronucleated polychromatic erythrocytes. PMID- 8709166 TI - [A photoallergenicity study of prulifloxacin (NM441) in guinea pigs]. AB - Photoallergenicity of prulifloxacin, a new antibacterial agent, was examined using guinea pigs compared with those of the other quinolone antibacterial drugs, ofloxacin (OFLX), lomefloxacin (LFLX), ciprofloxacin (CPFX), enoxacin (ENX) and nalidixic acid (NA). Prulifloxacin and other drugs were orally administered at minimal phototoxic doses 1 hr before UVA (18 J/cm2) irradiation. This photosensitization procedure was daily repeated for 5 days. On 16 days after the final sensitization, the animals were challenged with UVA (18 J/cm2) after the administration of correspondent substances at maximal non-phototoxic doses. Photoallergic reactions were induced by OFLX (40 mg/kg), LFLX (3 mg/kg), CPFX (170 mg/kg) and ENX (80 mg/kg), but were not observed in prulifloxacin (170 mg/kg) and NA (30 mg/kg). These results show that photoallergenicity of prulifloxacin were less severe than those of the other quinolone antibacterial drugs under the conditions of this study. PMID- 8709167 TI - [Renal toxicity of prulifloxacin (NM441) in rats]. AB - Renal toxicity of prulifloxacin, a new antibacterial agent, was investigated in rats of both sexes. The animals were given prulifloxacin orally for 28 days at a dose of 3000 mg/kg. Tubular nephrosis in which crystalline substances appeared primarily within tubules was observed from the second day of administration, and a large number of brown circular crystals were found in the urinary sediment from the first day of administration. Electron microscopic observation revealed a close resemblance of the ultrastructural characteristics between the intratubular crystalline substance and the urinary brown circular crystal, and the tubules were occasionally occluded by the crystalline substances. Infrared spectral analysis and X-ray microanalysis indicated that the brown circular crystal consisted of NM394, an active metabolite of prulifloxacin. These results suggested that NM394, which was filtered into the primary urine, may be precipitated as crystals on the process of water reabsorption in the tubules. And then most of the crystals would be washed out as crystalluria particles, and some of crystals retained and caused the obstructive uropathy. PMID- 8709168 TI - [Single-dose toxicity studies of prulifloxacin (NM441) in mice, rats and dogs and the active metabolite (NM394) in rats]. AB - Single-dose toxicity studies of prulifloxacin, a new antibacterial agent, were conducted in mice, rats and dogs. In addition, a single-dose toxicity study of (+/-)-6-fluoro-1-methyl-4-oxo-7- (1-piperazinyl)-4H-[1,3]thiazeto[3,2-a]quinoline 3-carboxylic acid (NM394), an active metabolite of prulifloxacin, was performed in rats. Prulifloxacin was administered orally, intraperitoneally (i.p.) or subcutaneously (s.c.) to mice and rats, and orally to dogs. NM394 was administered intravenously (i.v.) to rats. When prulifloxacin was administered orally or s.c., LD50 values were more than 5000 mg/kg in both sexes of mice and rats; when it was administered i.p., LD50 values were 1757 mg/kg in male mice, 1652 mg/kg in female mice, 915 mg/kg in male rats, and 1076 mg/kg in female rats. The lethal doses of this drug were more than 5000 mg/kg in both sexes of dogs by the oral route. The LD50 values of NM394 were 226 mg/kg in male rats and 238 mg/kg in female rats by the i.v. route. In mice, the major clinical signs observed following the administration of prulifloxacin were sedation, oligopnea, abnormal gait, piloerection, closed eye and tremor by the i.p. route and a scab at the site of injection by the s.c. route; in rats, decreased spontaneous locomotor activity by any of the three routes, oligopnea, lacrimation, hypothermia, piloerection and abnormal gait by the i.p. route, and a scab at the site of injection by the s.c. route; and in dogs, vomiting, reddening of the skin, and loose stool by the oral route. When NM394 was administered i.v., rats showed clonic convulsion and dyspnea. The site of injection was hyperemic, swollen and necrotic. Mice showed a decrease in body weight or an inhibition in weight gain when prulifloxacin was administered i.p. and rats showed the same effects when prulifloxacin or NM394 was administered by any of the above mentioned routes. Macroscopic findings detected following the i.p. administration of prulifloxacin in mice were pale color of the liver and spleen, thickening of the liver, and adhesion of intra-abdominal organs; and in rats, hydrothorax, congestion and edema of the lung, adhesion of intra-abdominal organs, swelling of the kidney accompanied by fine yellowish-white foci, and atrophy of the testis. When NM394 was administered i.v. to rats, congestion of the lung was macroscopically observed. PMID- 8709169 TI - [A 4-week oral toxicity study of prulifloxacin (NM441) in rats followed by a 4 week recovery test]. AB - A repeated dose toxicity study of prulifloxacin, a new antibacterial agent, was conducted in Sprague-Dawley rats. Male and female rats were given the test material orally for 4 weeks at doses of 0 (control), 30, 300 and 3000 mg/kg. After discontinuation of the treatment, a 4-week recovery test was also conducted. There was one case of death in the 3000 mg/kg group. Grayish green and soft feces, unkempt fur, transient deep respiration and decreased body weight gain were observed in the 3000 mg/kg group. Decreased food consumption and increased water intake were seen in the 300 and 3000 mg/kg groups. Ophthalmoscopic examination failed to show any abnormalities related to the treatment. In urinalysis, crystalline substance in the urinary sediments, cloudy urine and decreased Na+ excretion were observed in the 300 and 3000 mg/kg groups. Increased urine volume, lowered urine specific gravity and decreased K+ and Cl- excretions were seen in the 3000 mg/kg group. Hematologic examination showed decreased Hb, Ht, MCV and MCH and increased WBC in the 3000 mg/kg group. Blood chemical examination revealed increased BUN and decreased K+ and Cl- in the 3000 mg/kg group, and decreased K+ and gamma-globulin in the 300 mg/kg group. Pathological changes caused by the treatment were as follows. Cecal weight was increased in all dose groups. Cecal distention and swelling of its absorptive cells were seen in the 300 and 3000 mg/kg groups. In kidney, tubular nephrosis with crystalline substance was observed in the 300 and 3000 mg/kg groups, and its organ weight was increased in the 3000 mg/kg group. The above-mentioned changes were reversible except for decreased gamma-globulin, increased BUN and urine volume, and lowered urine specific gravity. Ulcer and small cavities associated with proliferation of fibrous tissue in the femoral articular cartilage were observed in the 3000 mg/kg group at the end of recovery period of 4 weeks. Plasma levels and urinary concentrations of active metabolite of the test material were increased in all dose groups with dose-related manner, whereby no sex difference was observed. No effects caused by the repeated dosing were seen in the plasma concentrations. Increased cecal weight in the 30 mg/kg group was considered to be attributable to the pharmacological effect of the test material. The results show that the NOAEL of prulifloxacin is 30 mg/kg for 4-week repeated dose toxicity in rats. PMID- 8709170 TI - [A 4-week oral toxicity study of prulifloxacin (NM441) in dogs followed by a 4 week recovery test]. AB - A repeated dose toxicity study of prulifloxacin, a new antibacterial agent, was conducted in beagle dogs. Male and female dogs were given the test material orally for 4 weeks at doses of 0 (control), 30, 150 and 750 mg/kg. After discontinuation of the treatment, a 4-week recovery test was also conducted. Feces containing white material were seen in the 150 and 750 mg/kg groups. Salivation, prone, lateral or sitting position, gait disturbance, and locomotor depression were observed in the 750 mg/kg group. In this dose group, decreased body weight and food and water consumptions were also observed. There were no treatment-related effects on survival. Ophthalmoscopic and electrocardiographic examinations and urinalysis failed to show any abnormalities related to the treatment. Hematologic examination showed decreased WBC in the 750 mg/kg group. Blood chemical examination revealed increased GPT and alpha 2-globulin in the 750 mg/kg group. Pathological changes caused by the treatment were as follows. Rarefaction of matrix, cavitations and erosions in humeral and femoral articular cartilages, and inflammatory cell infiltration in synovium were seen in the 150 and 750 mg/kg groups. Focal hemorrhage in synovium was also observed in the 750 mg/kg group. In kidney, regeneration of tubular epithelium, inflammatory cell infiltration, fibrosis and crystalline substance in the tubular lumen were observed in the 750 mg/kg group. The above-mentioned changes were satisfactorily reversible except for the changes in the humeral and femoral articular cartilages and in the kidney. Plasma levels and urinary concentrations of active metabolite of the test material were increased in all dose groups with dose-related manner, whereby no sex difference was observed. No effects caused by the repeated dosing were seen in the plasma concentrations. Toxicological findings were not observed in the 30 mg/kg group. The results show that the NOAEL of prulifloxacin is 30 mg/kg for 4-week repeated dose toxicity in dogs. PMID- 8709171 TI - [A 13-week oral toxicity study of prulifloxacin (NM441) in rats followed by a 5 week recovery test]. AB - A repeated dose toxicity study of prulifloxacin, a new antibacterial agent, was conducted in Sprague-Dawley rats. Male and female rats were given the test material orally for 13 weeks at doses of 0 (control), 30, 170 and 1000 mg/kg. After discontinuation of the treatment, a 5-week recovery test was also conducted. Salivation, soft feces, reduced body weight gain and increased water consumption were seen in the 1000 mg/kg group. There were no treatment-related effects on survival and food consumption. Ophthalmoscopic and hematologic examinations failed to show any abnormalities attributable to the treatment. Urinalysis revealed increased urine volume and decreased K+ excretion in the 1000 mg/kg group. Blood chemical examination showed increased BUN, decreased triglyceride, K+, Cl- and total protein in the 1000 mg/kg group. Pathological changes caused by the treatment were as follows. Renal tubular nephrosis with crystalline substance was observed in the 170 and 1000 mg/kg groups. Renal weight was increased and crystalline substance was noted in the lumen of the urinary bladder in the 1000 mg/kg group. Cecal distention with increased its organ weight was observed in all dose groups and swelling of its absorptive cells was seen in the 170 and 1000 mg/kg groups. Swelling of jejunal goblet cells was observed in the 1000 mg/kg group. In femoral articular cartilage, focal accumulation of chondrocytes, small cavities and proliferation of fibrous tissue were seen in the 170 and 1000 mg/kg groups. The above-mentioned changes were reversible except for renal tubular nephrosis and cecal distention with its increased organ weight, of which the degree and frequency, however, were lowered. The cecal distention in the 30 mg/kg group was considered to be attributable to the pharmacological effect of the test material. The results show that the NOAEL of prulifloxacin is 30 mg/kg for 13-week repeated dose toxicity in rats. PMID- 8709172 TI - Postpartum depression among adolescent mothers and adult mothers. AB - This study examined the extent to which adolescents are at increased risk for postpartum depression. Mothers, ranging in age from 15-37, comprised two samples: 77 adolescent mothers (15-20 years) and 151 adult mothers (21-37 years). Six weeks postpartum, 61% of the adolescent mothers and 37.7% of the adult mothers met Beck Depression Inventory criteria for mildly to severely depression. Adolescent mothers reported significantly higher perceived stress and postpartum depression, but lower social support and self-esteem than adult mothers. Among 21 symptoms of depression, adolescent mothers exhibited significantly higher cognitive-affective symptoms than adult mothers. Adolescent mothers reported higher levels of somatic symptoms than adult mothers, but did not reach statistical significance. Low self-esteem was the significant risk factor of postpartum depression for adolescent mothers. In addition to low self-esteem, high stress was significant risk factor of postpartum depression for adult mothers. PMID- 8709173 TI - Patients' needs and their coping strategies transition to cardiac surgery. AB - Little data are available concerning Taiwanese patients' needs and the factors contributing to their needs before cardiac surgery. The purpose of this study was to explore the components, nature and definition of patients' needs and their coping strategies of preparation for cardiac surgery during admission transition from patients' perspectives. The admission transition in this study means the patients' stay in the cardiovascular floor unit before surgery. Thirty adult patients (15 men, 15 women) having cardiac surgery were theoretically sampled from three hospitals in Taipei, Taiwan. Data were obtained through a semi structured interview with the investigator and were analyzed by employing the combination of strategies from grounded theory, qualitative content analysis, linking data, and constant comparison. Ninety percent of participants expressed two types of needs: certain- including met, unmet and expectant needs, and uncertain needs. Needs that participants believed to have been met were the needs for security, maintaining daily living activities, understanding the surgery and recovery process, the needs for optimal physical condition for surgery, receiving quality treatment, and coaching as to use of medical instruments. The unmet needs were the needs for maintaining daily activities, security, fulfilling unfinished family responsibilities, financial support, understanding the surgery and what to expect during the recovery process, freedom from pain, family members' companionship, and spiritual support. The expectant needs included those needs during the ICU and discharge-preparation recovery transitions, throughout the hospitalization and after recovery. The components, nature, definition, reasons, and suggestions for patients' needs and their coping strategies were delineated and discussed. Finally, a conceptual framework was developed to describe and depict this phenomenon. PMID- 8709174 TI - Familial supravalvular aortic stenosis: a report of two brothers. AB - We studied two brothers in a family with isolated congenital supravalvular aortic stenosis without Williams syndrome. Clinical, echocardiographic, hemodynamic and angiographic features of our cases were described. A severe long, narrowed supravalvular aortic stenosis involving the ascending aorta with proximal narrowing of the aortic branches and mild aortic insufficiency were found. To the best of our knowledge, this is the first report of this combination of lesions in the Orient. PMID- 8709176 TI - Clinical characteristics of and response to combination chemotherapy and subsequent application of international prognostic index in non-Hodgkin's lymphoma--an experience from a medical center in Southern Taiwan. AB - A retrospective analysis was performed of 117 non-Hodgkin's lymphoma (NHL) patients (72 male and 45 female, mean age 55 years) treated at NCKUH between July 1988 and December 1993. Of the 115 patients who could be classified by Ann Arbor staging system, 26 patients (22.2%) were in stage 1; 23 (19.7%) in stage 2; 29 (24.8%) in stage 3; and 37 (31.6%) in stage 4. According to the International Working Formulation, three patients (2.6%) were low grade NHL, 90 (76.9%) were intermediate, and 8 (6.8%) were high grade NHL. Histologically, diffuse large cell NHL accounted for 52.1% of cases, followed by 16.2% of cases exhibiting diffuse mixed NHL. Immunophenotype analysis was available in 95 cases, which revealed 76 (80%) cases exhibiting B-cell origin, 17 (18%) cases exhibiting diffuse mixed NHL. Immunophenotype analysis was available in 95 cases, which revealed 76 (80%) cases exhibiting B-cell origin, 17 (18%) cases exhibiting T cell origin and 2 (2%) cases were of null cell type. All patients underwent two groups of induction chemotherapy, either CHOP (Cyclophosphamide, Epirubicin, Oncovin, and Prednisolone), or "modified" COPBLAM (Cyclophosphamide, Epirubicin, Oncovin, and Prednisolone), or "modified" COPBLAM (Cyclophosphamide, Epirubicin, Oncovin, Vinblastine, Bleomycin, Procarbazine, and Prednisolone). Seventy-two cases treated through COPBLAM and 45 cases treated through CHOP were evaluated. The response rate (RR) to COPBLAM treatment was 72.2% and was 68.9% for the CHOP group (P = 0.51). The 5-year overall survival rate (OAS) was 44.1% for COPBLAM, versus 40% for CHOP (P = 0.15). The disease-free survival (DFS) was 72.6% at 63 months for COPBLAM and 58% at 51 months for CHOP (P = 0.16). Neither B cell nor T cell lineages of NHL showed any statistical difference in RR (P = 0.53, DFS (P = 0.58) or OAS (P = 0.97) to the different treatments. Using multiple logistic analysis, two independent factors, high LDH and advanced stage, were found to adversely affect the rate of complete remission. The application of the International Prognostic Index to our patients needs modification, which suggests the necessity of more evaluation before it can accurately be applied to all international series of NHL. PMID- 8709175 TI - Cytokeratin fragment 19 (CYFRA 21-1) as a tumor marker in non-small cell lung cancer. AB - To evaluate the diagnostic value of the serum cytokeratin 19 fragment (CYFRA 21 1) in bronchogenic carcinoma, we investigated the sera of 138 patients (58 with benign pulmonary disease and 80 with non-small cell lung cancer (NSCLC)) using immunoradiometric assay. The mean (SD) value of serum CYFRA 21-1 in NSCLC (13.26 (16.54)) was significantly higher than in benign lung diseases (1.74 (1.55)) (p < 0.0001). Sensitivity for CYFRA 21-1 (using 3.5 ng/ml, a cut-off value corresponding to a 95% specificity for benign pulmonary disease) in NSCL was 62%. Positive CYFRA 21-1 levels were significantly higher in 75% of patients with squamous cell carcinoma (n = 36) than in 53% with other NSCLC (n = 44) (p < 0.05). CYFRA 21-1 levels were significantly different between squamous cell carcinoma (17.28 (19.94)) and the other NSCLC (9.96 (12.44)) (P < 0.05). Elevated CYFRA 21-1 levels in patients with stage III and IV disease (n = 64, 18.19 (26.51)) were significantly higher than in stage I and II (n = 16, 4.41 (5.76)) (p < 0.02). The positive rate of CYFRA 21-1 in tumor stage I and II was only 37%. Our results indicate that CYFRA 21-1 may be a useful tumor marker in NSCLC, especially in squamous cell carcinoma. However, CYFRA 21-1 cannot be used for the diagnosis of early stage disease of NSCLC. CYFRA 21-1 may also contribute to the monitoring of NSCLC. PMID- 8709177 TI - Effect of maternal labor and mode of delivery on neutrophil actin response to N formylmethionyl-leucyl-phenylalanine in healthy neonates. AB - Diminished actin response to N-formyl-methionyl-leucyl-phenylalanine (FMLP) has been used to explain the impaired chemotaxis of neonatal neutrophils, but the effect of labor on this response has not been evaluated before. Therefore, we tested the hypothesis that labor stress may have an effect on actin response of cord blood neutrophils to FMLP. This response is compared by phallacidin stain of neutrophil F-actin and flow cytometry analysis among 3 groups of healthy neonates: group I, 16 vaginally delivered neonates; group II, 16 neonates delivered by elective cesarean section without labor; and group III, 16 neonates delivered by cesarean section after labor. In the group III, 10 infants were delivered by an emergency cesarean section because of fetal heart rate deceleration indicating fetal distress (one minute Apgar score < 7 in 5 cases [50%]), and the other 6 infants were delivered because of failure of progression of labor (one minute Apgar score < 7 in 2 cases [33.3%]). Stimulated relative F actin content of neutrophils in group III was increased compared with groups I and II. There was no such difference between neutrophils from infants of groups I and II. The results of this study indicate that fetal distress may enhance neutrophil actin response to FMLP, while labor or mode of delivery might not have such effect. PMID- 8709178 TI - The relationships between the radionuclide alveolar integrity study and the pulmonary function test. AB - This study evaluated the relationships between the alveolar integrity and the pulmonary function in 33 male subjects, including 18 COPD patients, 10 normal volunteers and 5 NIDDM patients. Alveolar integrity was evaluated by dynamic Tc 99m DTPA/HMPAO radioaerosol lung scintigraphies. The speed of radioaerosol clearance is represented as a slope from the lungs to the blood stream. Comprehensive pulmonary function tests included forced expiratory volume, flow volume loop, and diffusion capacity of a single breath CO method. The results showed the following: that (1) there are no correlations between the slopes and the parameters of pulmonary function tests, and (2) that the clearance of DTPA aerosols is faster than the clearance of HMPAO, and that these differences are significant. In conclusion, the radionuclide alveolar integrity study should be considered as a new method for evaluating lung function, and should be differentiated from traditional pulmonary function tests, and, secondly, lipophilic HMPAO aerosols are slower to clear than those of hydrophilic DTPA, which suggests at least two different clearance mechanisms for radioaerosols. PMID- 8709179 TI - Effect of body position on isometric arm lifting strength. AB - The purpose of this study was to examine the influence of three different body positions on isometric arm lifting strength. Thirty students aged between 20 and 27 years volunteered as subjects for the study. A force Evaluation and Testing System (FET 5000) was used to measure the arm lifting strength in the supine, sitting, and standing positions. Average force of each body position was determined by the mean of two trials. At the same time, electromyographic activity of bilateral biceps brachii muscles was measured during maximal voluntary isometric arm lifting. Repeated measures analysis of variance was used to analyze the effects of body positions on the amount of average force value and integrated electromyographic (IEMG) values during isometric arm lifting. The results showed that the highest mean lifting strength was recorded in the sitting position for both males and females. Significant difference in arm lifting strength was found between each body position. Differences in force were elicited more easily between the female and male subjects. However, the average IEMG values were not significantly correlated with the force produced in arm lifting. Nevertheless, this study supports the conclusion that body position did influence the production of isometric elbow flexors strength. We conclude, therefore, that body position must be considered during rehabilitation of upper extremities. PMID- 8709180 TI - [Study on technetium-99m in radiopharmaceuticals]. AB - The technetium field involving a new element of which chemistry has not been honed in the research during the century is particularly challenging, it is exciting and satisfying to see the successful development of a radiopharmaceutical agent which gives the clinical doctor new tools with which to assess function and in vivo biochemistry in nuclear medicine. The clinical application of technetium in nuclear medicine will parallel with the development of technetium chemistry in organic-, analytical-, medical- and biochemistry, and it is likely that useful new Tc-99m radiopharmaceuticals will rapidly make world wide progress in the future years. The design and synthetic modifications of ligand structure in organic chemistry will impart new biological properties to the technetium complexes. A structure characterization Tc-99m labelled radiopharmaceuticals is highly desirable in analytical chemistry. We take much interest in the structural characterization of Tc-99m complexes stems to establish a structure/in vivo activity relationship in medical chemistry. The current clinical trend is to use Tc-99 radiopharmaceuticals with Tc-99m "carrier added" experiments for studies of chemical structure and biochemical mechanism of action. Technical difficulties in the various labeling approaches to these bifunctional chelating agents (BCA) which seem promising are often encountered during the ached procedures. High priority is given to development of Tc-99 m radiopharmaceuticals for studies of substrate utilization and localization of specific biological recognition sites. New accomplishments in technetium radiopharmaceutical are likely to be reported in the near future. PMID- 8709181 TI - [Parasitic infections among wild rats from two areas of Kaohsiung]. AB - Wild rats including four strains were captured for examination of parasitic infections. The numbers were 101 from areas of Kaohsiung harbor and 98 from areas of Kaohsiung airport. Most of the examined wild rats were Rattus norvegicus and second most were Suncus murinus. The infection rates of intestinal helminths in rats of both areas did not display any significant difference. The overall infection rate was 29.6%. The infection rate was highest in R. norvegicus and the most frequently found parasite in rats was Hymenolepis diminuta. Moniliformis moniliformis and Angiostrongylus cantonensis were revealed in two separated R. norvegicus rats. The infection rates of intestinal helminths in R. norvegicus were significantly correlated with the size of rats. There was no find in microscopic observation of blood smear from these rats stained with Giemsa's dye. PMID- 8709182 TI - [Costand effectiveness assessment of laparoscopic cholecystectomy]. AB - "Cholecystectomy" is one of the ten "case payment" diseases announced in the National Health Insurance. Thirty-five consecutive laparoscopic cholecystectomies (LC), performed during the period July 1993 to December 1994, were compared to 35 age- and sex-matched patients undergoing open cholecystectomies (OC) during the same period. The results revealed that there were no significant differences in insurance status, indications, or abnormal results of EKG and liver function tests. The mean operative time was 124 +/- 46 min for LC, and 86 +/- 27 min for OC (p < 0.001), while the mean hospital stay was 7.4 +/- 2.8 days for LC, and 11.7 +/- 6.6 days for OC (p < 0.001). There were fewer patients using analgesics postoperatively within 24 hours for LC than for OC. The percentage of patients returned to work (or normal activity) within one week was higher for LC than for OC, but the difference was not statistically significant. Bile leakage was found in two LC cases (5.71%), while another two (57.1%) had wound infection in OC. Cost analysis showed that the cost of the operation room was NT$18,503 higher for LC (NT$27,643 against NT$9,140). Hospital charges were, on average, NT$53,721 for LC and NT$51,560 for OC (P > 0.05). The higher cost of LC was offset by a reduced length of hospital stay, so the hospital charges in the cases of both procedures were similar. As experience increases, the cost of LC will become lower. Moreover, LC minimizes the discomfort of patients, and patients can return to work earlier. From the perspective of the healthcare system, Laparoscopic Cholecystectomy is a safe, cost-effective technology. PMID- 8709183 TI - [Clinical manifestations and effects of IVGG in patients with Kawasaki disease]. AB - From Jan 1984 till Dec 1992, 293 patients--180 males and 113 females (M:F = 1.5:1)--with Kawasaki disease visited the Pediatric Department of Kaohsiung Medical College Hospital. The mean age from the total cases were 28.5 months (2 months-10 years). Fever was the most common clinical symptom, followed by dry cracked lips (93.5%), pharyngeal injection (92.8%) and conjunctivitis (90.5%). Only 62.6% of the total cases had cervical lymph node swelling of which the prevalence was higher than the previous report of the National Taiwan University Hospital in 1985. Two hundred and ninety-three cases were divided randomly into 3 groups according to the different treatment regimens. The first group of 199 cases whose coronary artery change occurred in 85 cases (42.7%), were treated with aspirin alone. The second group of 80 cases were treated with aspirin and IVGG 400 mg/kg/day for 5 consecutive days. The prevalence of coronary artery abnormalities was 22.5% (18/80). The third group of 14 cases were treated with aspirin and single high dose IVGG (2 gm/kg) for 10-12 hours. Coronary artery abnormalities occurred in 3 cases (21.4%). IVGG, initiated within 10 days of the onset of fever, in conjunction with aspirin decreased the prevalence of coronary artery dilatation and aneurysms significantly in comparison with treatment by the aspirin alone (p < 0.05). However, there was no difference in the prevalence of coronary aneurysm between the groups of single high dose and multiple doses, though the single high dose of IVGG can improve the clinical symptoms quickly and shorten the duration of hospitalization. PMID- 8709184 TI - [The practicability of preceptorship in the curriculum of medical education]. AB - Physicians who have graduated from traditional medical education curriculum focus on technological and organ system training, and are deficient in modern community health care concepts and skills. A preceptorship course integrates at various medical educational periods is one of the important goals at education reform in America. In this study, we attempt to ascertain the opinions of family physicians about the current family medicine curriculum and the practicability of preceptorship in Taiwan. The result can be a reference for future medical curriculum design. Two hundreds and thirty-five (40.8%) questionnaires were collected and analyzed. They showed most respondents were dissatisfied with the current curriculum of family medicine, and most agree that primary care physicians involvement would be helpful in producing efficient and well-qualified physicians. They suggested that preceptorship should best be first integrated in a basic knowledge course. We suggest that faculties who are responsible for curriculum design should evaluate the current curriculum and the appropriateness of preceptorship in Taiwan. Prior to the reform, faculty development and evaluation are the essential and crucial tasks. PMID- 8709185 TI - [A survey on sex knowledge and medical demand of the cases in Kaohsiung city adolescent health clinic]. AB - The purpose of this survey is to study the sex knowledge of adolescents, and to analyze their need in sex consultation and related health problem in Kaohsiung city. A total of 1,000 study subjects were recruited from May 1991 to May 1994. A self-administered questionnaire and medical records were used to obtain related information. The results were as follows: 1. The male to female ratio of study subjects was 1:2. Half of them were students, 44.1% had sexual experience, and the mean +/- standard deviation of age at first sex experience was 19.1 +/- 2.8 years. 2. The average percentage of correct knowledge was as low as 67.6%. It indicated an inadequacy of sex knowledge in adolescents. Females had better knowledge than males. Subjects who had graduated from senior high schools had better knowledge than those graduated from middle schools. No association was found between the score of sex knowledge and occupation. 3. 12.5% of study subjects believed that "masturbation" might cause neurasthenia, kidney malfunction and loss of memory, while 11.9% recognized that "nocturnal emission" might result in abnormal ejaculation, mental aberration and guilty feelings. Furthermore, 3.8% considered that kissing, hugging or sleeping together might cause pregnancy. 4. Most adolescents felt that "guidance on sex knowledge" was the major reason for them to visit the adolescent clinic and "obstetric and gynecological problems" was the second most important. 5. Males and females have the same prevalence of sex experience, and 50.6% of them did not practice contraception. 88.9% subjects had a high risk of getting pregnancy. 6. Males had a higher percentage of more than one heterosexual partner than females did. There was a higher percentage of people with a high education level having a fixed heterosexual partner than that of those with a low education level. PMID- 8709186 TI - [Compare stress and social support between adolescent and adult pregnant women during third trimester]. AB - With the change of society, adolescent pregnancy is the issue for the professionals to face and be concerned about. The results of the study can be utilized to improve the prenatal care for adolescents. A total of 90 adolescents and 90 adults during third trimester were recruited to participate in this study. Three methods were used to collect data: the Demographic Data Form, the Pregnancy Stress Rating Scale, and the Interpersonal Support Evaluation List. Primiparas showed a significantly higher score on stress from seeking safe passage for herself and her child through pregnancy, labor and delivering than multiparas. Adult pregnant women had a higher score on social support than did pregnant adolescents. Results of the stepwise multiple regression indicated that the best subset to predict stress from identifying maternal role was social support and education. The best subset to predict stress from seeking safe passage for herself and her child through pregnancy, labor and delivery was parity and education, the best predictor of stress from altering body structure and body function was education. PMID- 8709187 TI - [Unusual spontaneous dural cavernous sinus fistula between right external carotid artery and left cavernous sinus--a case report]. AB - A 64 year-old female had left exophthalmos which resulted from a spontaneous dural cavernous sinus fistula between the right external carotid artery and left cavernous sinus. The left cavernous sinus was fed directly from internal maxillary branches of right external carotid artery with the sparing of right cavernous sinus and intracavernous sinus. Symptoms and signs of dural cavernous sinus fistula developed slowly in the left eye only. Although this kind of spontaneou dural cavernous sinus fistula is very rare. The possibility of arterial feeding from contralateral carotid artery should not be ignored in any patient that is suffered from dural cavernous sinus fistula while angiographic study is performed. PMID- 8709188 TI - [Characteristics of dementia patients with hyperphagia]. AB - Research into dementia has tended to concentrate on memory loss and other cognitive impairment, but attention has recently turned to the associated psychiatric symptoms and behavioral abnormalities. Among them, increases in the amount eaten or hyperphagia are not uncommon in dementia. This problem is of clinical importance since it not only jeopardizes patient's health, but also is a cause of caregiver burden. The aim of this study was to assess the prevalence of hyperphagia in dementia. Additionally, possible mechanisms underlying this problem were reviewed and related to the demographic data, cognitive function test, radiological findings and other psychiatric symptoms. Out of a sample of 91 demented inpatients, hyperphagia was present in 30.8%. Hyperphagia was significantly associated with delusion, misidentification, aggressive behavior, hoarding behavior and pica phenomena. But these patients did not show difference in cognitive function, radiological abnormalities, repetitive behavior and depressed mood when compared with other demented patients. The possible etiologies were discussed, including physiological and psychological factors. Hyperphagia is a complex phenomenon, better understanding of the underlying pathogenesis may highlight specific pharmacological or behavioral methods for the management this troublesome behavior. PMID- 8709189 TI - Evidence for activation of the hepatitis B virus polymerase by binding of its RNA template. AB - The hepatitis B viruses replicate by reverse transcription of an RNA pregenome by using a virally encoded polymerase. A key early step in replication is binding of the polymerase to an RNA stem-loop (epsilon) of the pregenome; epsilon is both the RNA encapsidation signal and the origin of reverse transcription. Here we provide evidence that this interaction is also key to the development of enzymatic activity during biosynthesis of the polymerase. Duck hepatitis B virus polymerase expressed in Saccharomyces cerevisiae can synthesize DNA from epsilon containing RNAs and can also end label other small RNAs. Expression of functional polymerase in S. cerevisiae requires interaction between the polymerase and epsilon during or shortly after translation for it to develop any enzymatic activity; if epsilon is absent during expression, the polymerase is inactive on RNAs both with and without epsilon. Functional duck polymerase can also be produced by in vitro translation, and synthesis of the polymerase in the presence of epsilon induces resistance in the polymerase to proteolysis by papain, trypsin, and bromelain. Induction of the resistance is specific for epsilon sequences that can support RNA encapsidation and initiation of DNA synthesis. Induction of the resistance precedes initiation of DNA synthesis and is reversible by degradation of epsilon. These two sets of data (i) support a model in which binding of epsilon to the polymerase induces a structural alteration of the polymerase prior to the development of enzymatic activity and (ii) suggest that this alteration may be required for the polymerase to mature to an active form. PMID- 8709190 TI - The human immunodeficiency virus type 1 capsid p2 domain confers sensitivity to the cyclophilin-binding drug SDZ NIM 811. AB - Human immunodeficiency virus type 1 (HIV-1) specifically incorporates the host cell peptidyl-prolyl isomerase cyclophilin A into virions via contacts with the capsid (CA) domain of the Gag polyprotein Pr55gag. The immunosuppressant drug cyclosporin A and the nonimmunosuppressive cyclosporin A analog SDZ NIM 811 bind to cyclophilin A and inhibit its incorporation into HIV-1 virions. Both drugs inhibit the virion association of cyclophilin A and the replication of HIV-1 with a similar dose dependence. In contrast, these compounds are inactive against other primate lentiviruses which do not incorporate cyclophilin A, such as simian immunodeficiency virus (SIV). To locate determinants which confer sensitivity to SDZ NIM 811, we generated chimeric proviruses between HIV-1 and SIVmac. A hybrid SIVmac which has the CA-p2 domain of the Gag polyprotein replaced by the corresponding domain from HIV-1 replicated in an established CD4+ cell line and in human but not macaque peripheral blood mononuclear cells. The transfer of the HIV-1 CA-p2 domain to SIVmac led to the efficient incorporation of cyclophilin A, and SDZ NIM 811 effectively inhibited both the virion association of cyclophilin A and the spread of the hybrid virus in infected cultures. We conclude that the HIV-1 CA-p2 domain contains determinants which confer the necessity to interact with cyclophilin A for efficient virus replication. Furthermore, our data show that the CA-p2 domain can play a crucial role in species tropism. PMID- 8709191 TI - oriP is essential for EBNA gene promoter activity in Epstein-Barr virus immortalized lymphoblastoid cell lines. AB - During Epstein-Barr virus latent infection of B lymphocytes in vitro, six viral nuclear antigens (EBNAs) are expressed from one of two promoters, Cp or Wp, whose activities are mutually exclusive. Upon infection, Wp is initially active, followed by a switch to Cp for the duration of latency. In this study, the region upstream of Cp was analyzed for the presence of cis elements involved in regulating the activities of the EBNA gene promoters in established in vitro immortalized lymphoblastoid cell lines (LCLs). It was determined that oriP, the origin for episomal maintenance during latency, is essential for efficient transcription initiation from either Cp or Wp in LCLs, as well as in some Burkitt's lymphoma cell lines. Deletion of the EBNA2-dependent enhancer located upstream of Cp resulted in a ca. two- to fivefold reduction in Cp activity in the LCLs assayed. More extensive deletion of sequences upstream of Cp, including the EBNA2-dependent enhancer, resulted in nearly complete loss of Cp activity. This loss of activity was shown to correlate with deletion of two CCAAT boxes, a proximal CCAAT box located at bp -61 to -65 and a distal CCAAT box located at bp 253 to -257, upstream of Cp. Site-directed mutagenesis of these cis elements demonstrated that Cp activity is highly dependent on the presence of a properly positioned CCAAT box, with the dependence on the distal CCAAT box apparent only when the proximal CCAAT box was deleted or mutated. Deletion of the glucocorticoid response elements located at ca. bp -850 upstream of Cp did not result in a significant loss in activity. In general, deletions which diminished Cp activity resulted in induction of Wp activity, consistent with suppression of Wp activity by transcriptional interference from Cp. The identification of oriP and the EBNA2-dependent enhancer as the major positive cis elements involved in regulating Cp activity in LCL suggests that EBNA gene transcription is largely autoregulated by EBNA 1 and EBNA 2. PMID- 8709192 TI - Replication of murine coronavirus defective interfering RNA from negative-strand transcripts. AB - The positive-strand defective interfering (DI) RNA of the murine coronavirus mouse hepatitis virus (MHV), when introduced into MHV-infected cells, results in DI RNA replication and accumulation. We studied whether the introduction of negative-strand transcripts of MHV DI RNA would also result in replication. At a location downstream of the T7 promoter and upstream of the human hepatitis delta virus ribozyme domain, we inserted a complete cDNA clone of MHV DI RNA in reverse orientation; in vitro-synthesized RNA from this plasmid yielded a negative-strand RNA copy of the MHV DI RNA. When the negative-strand transcripts of the DI RNA were expressed in MHV-infected cells by a vaccinia virus T7 expression system, positive-strand DI RNAs accumulated in the plasmid-transfected cells. DI RNA replication depended on the expression of T7 polymerase and on the presence of the T7 promoter. Transfection of in vitro-synthesized negative-strand transcripts into MHV-infected cells and serial passage of virus samples from RNA-transfected cells also resulted in accumulation of the DI RNA. Positive-strand DI RNA transcripts were undetectable in sample preparations of the in vitro-synthesized negative-strand DI RNA transcripts, and DI RNA did not accumulate after cotransfection of a small amount of positive-strand DI RNA and truncated replication-disabled negative-strand transcripts; clearly, the DI RNA replicated from the transfected negative-strand transcripts and not from minute amounts of positive-strand DI RNAs that might be envisioned as artifacts of T7 transcription. Sequence analysis of positive-strand DI RNAs in the cells transfected with negative-strand transcripts showed that DI RNAs maintained the DI-specific unique sequences introduced within the leader sequence. These data indicated that positive-strand DI RNA synthesis occurred from introduced negative strand transcripts in the MHV-infected cells; this demonstration, using MHV, of DI RNA replication from transfected negative-strand DI RNA transcripts is the first such demonstration among all positive-stranded RNA viruses. PMID- 8709194 TI - The rev-responsive element negatively regulates human immunodeficiency virus type 1 env mRNA expression in primate cells. AB - The human immunodeficiency virus type 1 (HIV-1) Rev protein mediates the accumulation of unspliced and singly spliced viral transcripts within the cytoplasm of infected cells, late in the infection cycle, leading to the expression of the viral structural proteins, Gag, Pol, and Env. Rev binds to a complex RNA structure, the Rev-responsive element (RRE), present in all Rev responsive viral transcripts, relieving their nuclear sequestration. The precise mechanism by which RRE-containing transcripts are retained within the nucleus in the absence of Rev protein is not well understood. We previously demonstrated that the RRE alone plays a crucial role in the nuclear retention of RRE containing env transcripts in stably transfected Drosophila cells. Here we extend our previous observations and demonstrate that the RRE is a principal determinant of nuclear retention for envelope transcripts in primate cells and, in particular, human CD4+ T cells. PMID- 8709193 TI - Transdominant mutants of I kappa B alpha block Tat-tumor necrosis factor synergistic activation of human immunodeficiency virus type 1 gene expression and virus multiplication. AB - The human immunodeficiency virus type 1 (HIV-1) long terminal repeat (LTR) contains two binding sites for the NF-kappa B/Rel family of transcription factors which are required for the transcriptional activation of viral genes by inflammatory cytokines such as tumor necrosis factor alpha (TNF-alpha) and interleukin-1. In the present study, we examined the effect of transdominant mutants of I kappa B alpha on the synergistic activation of the HIV-1 LTR by TNF alpha and the HIV-1 transactivator, Tat, in Jurkat T cells. The synergistic induction of HIV-1 LTR-driven gene expression represented a 50- to 70-fold stimulation and required both an intact HIV-1 enhancer and Tat-TAR element interaction, since mutations in Tat protein (R52Q, R53Q) or in the bulge region of the TAR element that eliminated Tat binding to TAR were unable to stimulate LTR expression. Coexpression of I kappa B alpha inhibited Tat-TNF-alpha activation of HIV LTR in a dose-dependent manner. Transdominant forms of I kappa B alpha, mutated in critical serine or threonine residues required for inducer mediated (S32A, S36A) and/or constitutive (S283A, T291A, T299A) phosphorylation of I kappa B alpha were tested for their capacity to block HIV-1 LTR transactivation. I kappa B alpha molecules mutated in the N-terminal sites were not degraded following inducer-mediated stimulation (t1/2, > 4 h) and were able to efficiently block HIV-1 LTR transactivation. Strikingly, the I kappa B alpha (S32A, S36A) transdominant mutant was at least five times as effective as wild type I kappa B alpha in inhibiting synergistic induction of the HIV-1 LTR. This mutant also effectively inhibited HIV-1 multiplication in a single-cycle infection model in Cos-1 cells, as measured by Northern (RNA) blot analysis of viral mRNA species and viral protein production. These experiments suggest a strategy that may contribute to inhibition of HIV-1 gene expression by interfering with the NF-kappa B/Rel signaling pathway. PMID- 8709195 TI - The zinc finger of nucleocapsid protein of Friend murine leukemia virus is critical for proviral DNA synthesis in vivo. AB - Nucleocapsid protein NCp10 of murine leukemia virus (MuLV) is encoded by the 3' domain of gag and contains a zinc finger of the form Cys-X2-Cys-X4-His-X4-Cys flanked by basic amino acids. In the course of virus assembly, NCp10 is necessary for core formation, and the zinc finger flanked by the basic residues is required for the packaging of the genomic RNA dimer. In vitro, NCp10 exhibits strong nucleic acid binding and annealing activities that appear to be critical for virus infectivity since NCp10 promotes dimerization of the viral RNA containing the E/DLS packaging-dimerization signal and annealing of replication primer tRNA(Pro) to the initiation site of reverse transcription (PBS). Recent in vitro studies have suggested that NCp10 may also play a role in proviral DNA synthesis. To investigate the function of NCp10 in proviral DNA synthesis in vivo, we developed a simple and convenient genetic packaging system consisting of two DNA constructs expressing the packaging components gag-pol and env of Friend MuLV and a Moloney MuLV-based lacZ vector with either the MuLV E+ or a rat VL30 E packaging signal. This system allowed us to examine the consequences of a set of mutations in NCp10 on a single round of recombinant virus replication. Most mutations in the N- or C-terminal domain of NCp10 do not significantly alter infectivity, while those in the zinc finger drastically impair infectivity. Analysis of the viral RNA content in virions showed that all mutations in the zinc finger decrease but do not abolish packaging of the recombinant genome. Interestingly enough, mutation of Y-28 to S (mutation Y28S) in the zinc finger results in RNA packaging at a level similar to that observed upon deletion of three prolines and three arginines in the C-terminal domain of NCp10 (mutant delta PR3). However, mutant Y28S is noninfectious while mutant delta PR3 is only threefold less infectious than the wild-type virus, which prompted us to examine the role of NCp10 protein in proviral DNA synthesis in vivo using these nucleocapsid mutants. PCR amplification was used to analyze viral DNA synthesized in newly infected cells, and results indicate that the Y28S zinc finger mutation impairs reverse transcription, thus suggesting that the nucleocapsid protein zinc finger plays a key role in proviral DNA synthesis in vivo. PMID- 8709196 TI - Efficient lysis of human immunodeficiency virus type 1-infected cells by cytotoxic T lymphocytes. AB - Numerous studies of human immunodeficiency virus type 1 (HIV-1)-specific cytotoxic T lymphocytes (CTL) have examined their ability to recognize B-cell lines expressing recombinant HIV-1 proteins, but relatively few data regarding the lysis of HIV-1-infected cells by CTL are available. We studied the ability of HIV-1-specific CTL clones of defined epitope specificity and HLA restriction to lyse infected CD4+ cells at serial time points following infection. CD4+ cell lines were acutely infected with HIV-1 IIIB at a high multiplicity of infection, and the kinetics of cell lysis were examined and compared with the kinetics of viral replication. Intracellular HIV-1 p24 expression was detected by 1 to 2 days after infection, reaching over 98% positive cells by day 4. Recognition of the infected cells by HLA A2- and B14-restricted CTL clones closely paralleled intracellular p24 expression and preceded peak virion production. The maximal levels of lysis with Gag-, reverse transcriptase-, and envelope-specific clones were different, however. The Gag- and envelope-specific clones lysed infected cells at levels equivalent to peptide-sensitized controls, whereas lysis by the reverse transcriptase-specific clones plateaued at a lower level. Peptide titration curves indicated that this effect was not due to differences in sensitivity to the cognate epitopes for the different clones. Although HIV-1 infection induced an approximately 50% decrease in class I HLA expression on the surface of infected cells, lysis by CTL clones was unaffected. These studies indicate that HIV-1-specific CTL can efficiently lyse HIV-1-infected CD4+ cells and suggest that the partial downregulation of class I molecules in infected cells does not significantly affect recognition by CTL. PMID- 8709197 TI - Epitopes exposed on hepatitis delta virus ribonucleoproteins. AB - A total of 17 antibodies, raised in several nonhuman species and specific for different regions on the delta antigen (delta Ag), were used to map, via immunoprecipitation, those domains exposed on the surface of the viral ribonucleoprotein (RNP). These studies showed that the domains for the nuclear localization signal and the C-terminal extension, unique to the large form of delta Ag, are exposed. Also exposed is the C-terminal region of the small form of delta Ag. In contrast, reactivity was not found with the coiled-coil domain needed for protein dimerization. When the hepatitis delta virus (HDV) RNA was released by treatment of viral RNP with vanadyl ribonucleoside complexes, no change in the pattern of delta Ag epitope presentation was detected, consistent with the interpretation that a multimeric protein structure persists in the absence of RNA. These RNP studies have implications not only for understanding of the process of HDV assembly but also for evaluation of the immune responses of an infected host to HDV replication. PMID- 8709198 TI - Expression of tissue factor is increased in astrocytes within the central nervous system during persistent infection with borna disease virus. AB - Persistent tolerant infection of rats with borna disease virus (BDV) results in a central nervous system (CNS) disease characterized by behavioral abnormalities. These disorders occur without inflammation and widespread cytolysis in the CNS. Therefore, mechanisms other than virally induced destruction of brain cells may explain the CNS disturbance caused by BDV. Previously, we have shown that astrocytes in the CNS express tissue factor (TF). TF functions as the primary cellular initiator of the coagulation protease cascades, resulting in the generation of the protease thrombin. Proteases and their inhibitors play important roles in the development and physiology of the CNS, and altered protease activity has been implicated in the pathophysiology of various neurological diseases. Here, we present evidence that TF expression in the brain is markedly increased during persistent infection with BDV. Persistent infection of cultured astrocytes with BDV also increased TF expression as a result of both increased transcription of the TF gene and stabilization of TF mRNA. We speculate that increased TF expression within the brain parenchyma may lead to increased protease activity in the CNS and contribute to virus-mediated CNS functional impairment by affecting neural cell interactions. PMID- 8709199 TI - Effect of human immunodeficiency virus type 1 protein R (vpr) gene expression on basic cellular function of fission yeast Schizosaccharomyces pombe. AB - The human immunodeficiency virus type 1 (HIV-1) Vpr protein affects cell morphology and prevents proliferation of human cells by induction of cell cycle G2 arrest. In this study, we used the fission yeast Schizosaccharomyces pombe as a model system to investigate the cellular effects of HIV-1 vpr gene expression. The vpr gene was cloned into an inducible fission yeast gene expression vector and expressed in wild-type S. pombe cells, and using these cells, we were able to demonstrate the specific Vpr-induced effects by induction and suppression of vpr gene expression. Induction of HIV-1 vpr gene expression affected S. pombe at the colonial, cellular, and molecular levels. Specifically, Vpr induced small-colony formation, polymorphic cells, growth delay, and cell cycle G2 arrest. Additionally, Vpr-induced G2 arrest appeared to be independent of cell size and morphological changes. The cell cycle G2 arrest correlated with increased phosphorylation of p34cdc2, suggesting negative regulation of mitosis by HIV-1 Vpr. Treatment of Vpr-induced cell with a protein phosphatase inhibitor, okadaic acid, transiently suppressed cell cycle arrest and morphological changes. This observation implicates possible involvement of protein phosphatase(s) in the effects of Vpr. Together, these data showed that the HIV-1 Vpr-induced cellular changes in S. pombe are similar to those observed in human cells. Therefore, the S. pombe system is suited for further investigation of the HIV-1 vpr gene functions. PMID- 8709200 TI - Uptake of duck hepatitis B virus into hepatocytes occurs by endocytosis but does not require passage of the virus through an acidic intracellular compartment. AB - The infectious entry pathway of duck hepatitis B virus (DHBV) was investigated with primary duck hepatocytes. Virus uptake was measured by a selective PCR technique which allows for the detection of a successful infection without the need for viral replication or gene expression. To test whether DHBV uptake occurs by endocytosis, the effects of energy depletion were analyzed. The requirement for an acidic intracellular pH was tested with the lysosomotropic agent ammonium chloride. The data show that energy depletion prevents the uptake of DHBV into primary hepatocytes whereas ammonium chloride has no effect. From these data, we conclude that DHBV is taken up by its host cells by endocytosis. However, in contrast to that of most other enveloped viruses, escape of DHBV from the endocytotic route does not depend on an acidic intracellular compartment. PMID- 8709201 TI - Trypsin activation pathway of rotavirus infectivity. AB - The infectivity of rotaviruses is increased by and most probably is dependent on trypsin treatment of the virus. This proteolytic treatment specifically cleaves VP4, the protein that forms the spikes on the surface of the virions, to polypeptides VP5 and VP8. This cleavage has been reported to occur in rotavirus SA114fM at two conserved, closely spaced arginine residues located at VP4 amino acids 241 and 247. In this work, we have characterized the VP4 cleavage products of rotavirus SA114S generated by in vitro treatment of the virus with increasing concentrations of trypsin and with proteases AspN and alpha-chymotrypsin. The VP8 and VP5 polypeptides were analyzed by gel electrophoresis and by Western blotting (immunoblotting) with antibodies raised to synthetic peptides that mimic the terminal regions of VP4 generated by the trypsin cleavage. It was shown that in addition to arginine residues 241 and 247, VP4 is cleaved at arginine residue 231. These three sites were found to have different susceptibilities to trypsin, Arg-241 > Arg-231 > Arg-247, with the enhancement of infectivity correlating with cleavage at Arg-247 rather than at Arg-231 or Arg-241. Proteases AspN and alpha chymotrypsin cleaved VP4 at Asp-242 and Tyr-246, respectively, with no significant enhancement of infectivity, although this enhancement could be achieved by further treatment of the virus with trypsin. The VP4 end products of trypsin treatment were a homogeneous VP8 polypeptide comprising VP4 amino acids 1 to 231 and a heterogeneous VP5, which is formed by two polypeptide species (present at a ratio of approximately 1:5) as a result of cleavage at either Arg 241 or Arg-247. A pathway for the trypsin activation of rotavirus infectivity is proposed. PMID- 8709202 TI - Conditional human immunodeficiency virus type 1 protease mutants show no role for the viral protease early in virus replication. AB - The human immunodeficiency virus type 1 protease plays a critical role in the proteolytic processing of precursor polyproteins during virion maturation. Contradictory evidence has been obtained for a possible role for the protease early after infection, i.e., during DNA synthesis and/or integration. We have reexamined this question by using conditional mutants of the protease. In one set of experiments, protease mutants that confer a temperature-sensitive phenotype for processing were used to assess the need for protease activity early after infection. No significant difference from results with wild-type virus was seen when infections were carried out at either 35 or 40 degrees C. In a separate set of experiments, infections were carried out in the presence of a protease inhibitor. In this case, both wild-type virus and a drug-resistant variant were used, the latter as a control to ensure a specific effect of the inhibitor. Infection with either virus was not inhibited at drug concentrations that were up to 10-fold higher than those needed to inhibit intracellular processing by the viral protease. The results obtained by both of these experimental protocols provide evidence that the human immunodeficiency virus type 1 protease does not play a role early after infection. PMID- 8709203 TI - Effects of a naturally occurring mutation in the hepatitis B virus basal core promoter on precore gene expression and viral replication. AB - The basal core promoter (BCP) of hepatitis B virus (HBV) controls the transcription of both the precore RNA and the core RNA. The precore RNA codes for the secreted e antigen, while the core RNA codes for the major core protein and the DNA polymerase and also is the pregenomic RNA. The double mutation of nucleotides 1762 and 1764 in the BCP from A and G to T and A, respectively, is frequently observed in HBV sequences isolated from chronic patients. Several papers have reported conflicting results regarding whether this double mutation is important for e antigen expression. In order to address this issue, we have introduced this double mutation into the HBV genome and studied its effects on HBV gene expression and replication. Our results indicate that the mutated BCP can no longer bind a liver-enriched transcription factor(s) and that the transcription of only precore RNA and, consequently, the expression of e antigen were reduced. The reduction of precore gene expression was accompanied by an increase in progeny virus production. This increase was found to occur at or immediately prior to the encapsidation of the pregenomic RNA. Thus, the results of our in vitro study resolve the discrepancy of previous clinical observations and indicate that this double mutation suppresses but does not abolish the e antigen phenotype. The implications of these findings in the pathogenesis of HBV are discussed. PMID- 8709204 TI - The CR1 and CR3 domains of the adenovirus type 5 E1A proteins can independently mediate activation of ATF-2. AB - The adenovirus 12S E1A protein can stimulate the activity of the c-jun promoter through a conserved region 1 (CR1)-dependent mechanism. The effect is mediated by two AP-1/ATF-like elements, jun1 and jun2, that preferentially bind c-Jun-ATF-2 heterodimers. In this study, we show that the ATF-2 component of the c-Jun-ATF-2 heterodimer is the primary target for 12S E1A: 12S E1A can enhance the transactivating activity of the N terminus of ATF-2 when fused to a heterologous DNA-binding domain, whereas the transactivating activity of the c-Jun N terminus is not significantly affected. Activation of the ATF-2 N terminus by 12S E1A is dependent on CR1. In the context of the 13S E1A protein, CR1 and CR3 can both contribute to activation of ATF-2, and their relative contributions are dependent on the cell type. In contrast to activation of ATF-2 by stress-inducing agents, CR1-dependent activation of ATF-2 was found not to depend strictly on the presence of threonines 69 and 71 in the N terminus of ATF-2, which are targets for phosphorylation by stress-activated protein kinases (SAPKs). In agreement with this observation, we did not observe phosphorylation of threonines 69 and 71 or constitutively enhanced SAPK activity in E1A- plus E1B-transformed cell lines. These data suggest that CR1-dependent activation of ATF-2 by 12S E1A does not require phosphorylation of threonines 69 and 71 by SAPK. PMID- 8709205 TI - Characterization of a myxoma virus-encoded serpin-like protein with activity against interleukin-1 beta-converting enzyme. AB - A genomic library of myxoma virus (MV) DNA, a leporipoxvirus that causes myxomatosis, was constructed and screened by in vitro transcription-translation. A clone was selected on the basis of its strong reactivity with MV antiserum. Analysis of the corresponding DNA sequence and the deduced amino acid sequence revealed an open reading frame coding for a 34-kDa protein with strong homologies to members of the serpin superfamily. The gene encoding this new protein, called serp2, was localized on the MV genome. Interestingly, this gene is deleted in an attenuated strain. We constructed a baculovirus vector to produce recombinant Serp2 protein and raised specific antisera that allowed the characterization of Serp2 expression during the MV cycle. The biological relevance of this new serpin from MV was monitored, and it was shown that Serp2 could inhibit human interleukin-1 beta-converting enzyme activity. PMID- 8709206 TI - Analysis of early region 3 mutants of mouse adenovirus type 1. AB - Early region 3 (E3) of mouse adenovirus type 1 has the potential to produce three proteins which have identical amino termini but unique carboxy-terminal sequences. Three recombinant deletion viruses were constructed so that each could produce only one of the three E3 proteins. A fourth mutant that should produce no E3 proteins was also constructed. These recombinants were able to grow in mouse 3T6 cells and produced wild-type levels of viral mRNAs and proteins except for those specifically deleted by the mutations. Early mRNA production from the mutant viruses was analyzed by reverse transcriptase PCR and confirmed that each deletion mutant would be able to produce only one of the three E3 proteins. Late mRNA production was analyzed by Northern (RNA) blotting and found to be similar in wild-type and mutant viruses. Capsid morphology was unaltered in the mutant viruses as seen by electron microscopy. Immunoprecipitation of E3 proteins from infections of mouse 3T6 cells using an antiserum specific for all three E3 proteins was used to examine the effect of the introduced mutations on protein expression. Two mutants produced only one class of E3 protein as predicted from their specific mutations and mRNA expression profiles. One mutant virus failed to produce any detectable E3 proteins. The predicted E3-null mutant was found to be leaky and could produce low levels of E3 proteins. Outbred Swiss mice were infected with the E3 mutant viruses to determine if the E3 proteins have an effect on the pathogenicity of the virus in mice. All of the mutants showed decreased pathogenicity as determined by increased 50% lethal doses, indicating that the proteins of the E3 region are important determinants of the pathogenesis of mouse adenovirus in its natural host. PMID- 8709207 TI - In vitro generation and type-specific neutralization of a human papillomavirus type 16 virion pseudotype. AB - We report a system for generating infectious papillomaviruses in vitro that facilitates the analysis of papillomavirus assembly, infectivity, and serologic relatedness. Cultured hamster BPHE-1 cells harboring autonomously replicating bovine papillomavirus type 1 (BPV1) genomes were infected with recombinant Semliki Forest viruses that express the structural proteins of BPV1. When plated on C127 cells, extracts from cells expressing L1 and L2 together induced numerous transformed foci that could be specifically prevented by BPV neutralizing antibodies, demonstrating that BPV infection was responsible for the focal transformation. Extracts from BPHE-1 cells expressing L1 or L2 separately were not infectious. Although Semliki Forest virus-expressed L1 self-assembled into virus-like particles (VLPs), viral DNA was detected in particles only when L2 was coexpressed with L1, indicating that genome encapsidation requires L2. Expression of human papillomavirus type 16 (HPV16) L1 and L2 together in BPHE-1 cells also yielded infectious virus. These pseudotyped virions were neutralized by antiserum to HPV16 VLPs derived from European (114/K) or African (Z-1194) HPV16 variants but not by antisera to BPV VLPs, to a poorly assembling mutant HPV16 L1 protein, or to VLPs of closely related genital HPV types. Extracts from BPHE-1 cells coexpressing BPV L1 and HPV16 L2 or HPV16 L1 and BPV L2 were not infectious. We conclude that (i) mouse C127 cells express the cell surface receptor for HPV16 and are able to uncoat HPV16 capsids; (ii) if a papillomavirus DNA packaging signal exists, then it is conserved between the BPV and HPV16 genomes; (iii) functional L1-L2 interaction exhibits type specificity; and (iv) protection by HPV virus-like particle vaccines is likely to be type specific. PMID- 8709208 TI - Local and distant sequences are required for efficient readthrough of the barley yellow dwarf virus PAV coat protein gene stop codon. AB - Many viruses use stop codon readthrough as a strategy to produce extended coat or replicase proteins. The stop codon of the barley yellow dwarf virus (PAV serotype) coat protein gene is read through at a low rate. This produces an extended polypeptide which becomes part of the virion. We have analyzed the cis acting sequences in the barley yellow dwarf virus PAV genome required for this programmed readthrough in vitro in wheat germ extracts and reticulocyte lysates and in vivo in oat protoplasts. Two regions 3' to the stop codon were required. Deletion of sections containing the first 5 of the 16 CCN NNN repeats located 3' of the stop codon greatly reduced readthrough in vitro and in vivo. Surprisingly, readthrough also required a second, more distal element that is located 697 to 758 bases 3' of the stop codon within the readthrough open reading frame. This element also functioned in vivo in oat protoplasts when placed more than 2 kb from the coat protein gene stop in the untranslated region following a GUS reporter gene. This is the first report of a long-range readthrough signal in viruses. PMID- 8709209 TI - Various modes of basic helix-loop-helix protein-mediated regulation of murine leukemia virus transcription in lymphoid cell lines. AB - The transcriptionally regulatory regions of the lymphomagenic Akv and SL3-3 murine leukemia retroviruses (MLVs) contain two types of E-box consensus motifs, CAGATG. One type, EA/S, is located in the upstream promoter region, and the other, E(gre), is located in a tandem repeat with enhancer properties. We have examined the requirements of the individual E-boxes in MLV transcriptional regulation. In lymphoid cell lines only, the E(gre)-binding protein complexes included ALF1 or HEB and E2A basic helix-loop-helix proteins. Ectopic ALF1 and E2A proteins required intact E(gre) motifs for mediating transcriptional activation. ALF1 transactivated transcription of Akv MLV through the two E(gre) motifs equally, whereas E2A protein required the promoter-proximal E(gre) motif. In T- and B-cell lines, the E(gre) motifs were of major importance for Akv MLV transcriptional activity, while the EA/S motif had some effect. In contrast, neither E(gre) nor EA/S motifs contributed pronouncedly to Akv MLV transcription in NIH 3T3 cells lacking DNA-binding ALF1 or HEB and E2A proteins. The Id1 protein was found to repress ALF1 activity in vitro and in vivo. Moreover, ectopic Id1 repressed E(gre)-directed but not EA/S-directed MLV transcription in lymphoid cell lines. In conclusion, E(gre) motifs and interacting basic helix loop-helix proteins are important determinants for MLV transcriptional activity in lymphocytic cell lines. PMID- 8709210 TI - Requirements for kissing-loop-mediated dimerization of human immunodeficiency virus RNA. AB - Sequences from the 5' end of type 1 human immunodeficiency virus RNA dimerize spontaneously in vitro in a reaction thought to mimic the initial step of genomic dimerization in vivo. Dimer initiation has been proposed to occur through a "kissing-loop" interaction involving a specific RNA stem-loop element designated SL1: the RNA strands first interact by base pairing through a six-base GC-rich palindrome in the loop of SL1, whose stems then isomerize to form a longer interstrand duplex. We now report a mutational analysis aimed at defining the features of SL1 RNA sequence and secondary structure required for in vitro dimer formation. Our results confirm that mutations which destroy complementarity in the SL1 loop abolish homodimer formation, but that certain complementary loop mutants can heterodimerize. However, complementarity was not sufficient to ensure dimerization, even between GC-rich loops, implying that specific loop sequences may be needed to maintain a conformation that is competent for initial dimer contact; the central GC pair of the loop palindrome appeared critical in this regard, as did two or three A residues which normally flank the palindrome. Neither the four-base bulge normally found in the SL1 stem nor the specific sequence of the stem itself was essential for the interaction; however, the stem structure was required, because interstrand complementarity alone did not support dimer formation. Electron microscopic analysis indicated that the RNA dimers formed in vitro morphologically resembled those isolated previously from retroviral particles. These results fully support the kissing-loop model and may provide a framework for systematically manipulating genomic dimerization in type 1 human immunodeficiency virus virions. PMID- 8709211 TI - Epstein-Barr virus EBNA3A and EBNA3C proteins both repress RBP-J kappa-EBNA2 activated transcription by inhibiting the binding of RBP-J kappa to DNA. AB - Following infection by Epstein-Barr virus (EBV), the production of viral nuclear proteins EBNA1, EBNA2, EBNA3A, and EBNA3C and the viral membrane protein LMP1 is essential for the permanent proliferation of primary B lymphocytes to occur. Among these, the transcription factor EBNA2 is central to the immortalizing process, since it activates not only the transcription of all the EBNA proteins and LMP1, TP1, and TP2 but also certain cellular genes. EBNA2 is targeted to its DNA-responsive elements through direct interaction with the DNA-binding cellular repressor RBP-J kappa. In a transient-expression assay, the EBNA2-activated transcription was found to be downregulated by EBNA3A, EBNA3B, and EBNA3C. However, since it has been reported that EBNA3C, but not EBNA3A, directly contacts RBP-J kappa in vitro, these proteins appear to repress through different mechanisms. Here, we report for the first time that EBNA3A and EBNA3C both stably interact with RBP-J kappa and most probably repress EBNA2-activated transcription by destabilizing the binding of RBP-J kappa to DNA. PMID- 8709212 TI - Directional budding of human immunodeficiency virus from monocytes. AB - Time-lapse cinematography revealed that activated human immunodeficiency virus (HIV)-infected monocytes crawl along surfaces, putting forward a leading pseudopod. Scanning electron micrographs showed monocyte pseudopods associated with spherical structures the size of HIV virions, and transmission electron micrographs revealed HIV virions budding from pseudopods. Filamentous actin (F actin) was localized by electron microscopy in the pseudopod by heavy meromyosin decoration. Colocalization of F-actin and p24 viral antigen by light microscopy immunofluorescence indicated that F-actin and virus were present on the same pseudopod. These observations indicate that monocytes produce virus from a leading pseudopod. We suggest that HIV secretion at the leading edges of donor monocytes/macrophages may be an efficient way for HIV to infect target cells. PMID- 8709213 TI - Human immunodeficiency virus type 1 drug susceptibility during zidovudine (AZT) monotherapy compared with AZT plus 2',3'-dideoxyinosine or AZT plus 2',3' dideoxycytidine combination therapy. The protocol 34,225-02 Collaborative Group. AB - Human immunodeficiency virus type 1 (HIV-1) isolates obtained prior to and during a combination therapy trial comparing zidovudine (AZT; 3'-azidothymidine) monotherapy with AZT plus 2',3'-dideoxyinosine (ddI) or AZT plus 2',3' dideoxycytidine (ddC) were assessed for the development of drug resistance. Drug susceptibility was measured by using two different phenotypic assays, one that requires infection of peripheral blood mononuclear cells with HIV-1 isolated from cocultures and a second based on infection of HeLa CD4+ cells with recombinant virus containing the reverse transcriptase (RT) of the clinical isolate. In addition, genotypic assessment of resistance was obtained by DNA sequencing of the RT coding region. No difference in the development of AZT resistance was noted in isolates from individuals receiving AZT monotherapy or combination therapy. However, a low frequency of ddI or ddC resistance was seen in isolates from the combination arms, which may at least partially explain the enhanced efficacy observed with these drug combinations compared with monotherapy. It was noted from DNA sequencing that a relatively high frequency of the nonnucleoside RT inhibitor resistance mutation, codon 181 changed from encoding Tyr to encoding Cys, was present in some isolates both before and during nucleoside analog combination therapy. Since these patients were unlikely to have access to nonnucleoside RT inhibitors, it is probable that this mutation preexisted at a reasonable level in the wild-type virus population. Comparisons of the AZT susceptibility assays indicated a good correlation between the phenotypic and genotypic determinations. However, direct numerical comparisons between the phenotypic assays were not reliable, suggesting that valid comparisons of different resistance data sets will require the use of the same assay procedure. PMID- 8709214 TI - Significance of amino acid variation at human immunodeficiency virus type 1 reverse transcriptase residue 210 for zidovudine susceptibility. AB - Amino acid variation at reverse transcriptase (RT) codon 210 (generally Leu-210 to Trp [L210W], TTG-->TGG) is occasionally detected after the initiation of azidothymidine (AZT) therapy. The impact of this variation on AZT resistance and viral replication was addressed by four different approaches. The frequency and genetic background of the L210W mutation in vivo were assessed by analyzing sera of AZT-naive and AZT-experienced patients by RT-PCR and DNA sequencing. The degree of AZT resistance (50% infective concentration [IC50]) of recombinant viruses constructed by using the RT of 21 clinical isolates was stratified by the presence or absence of the 210 mutation. The AZT IC50S of a panel of mutant viruses (with or without W-210) constructed by site-directed mutagenesis in an HXB2 background were assayed by using a HeLa CD4 plaque reduction assay. Finally, the effect of the 210 mutation on viral replication was assessed by replication competition of an AZT-resistant virus, RTMN (L-41/Y-215), and RTMN with the W-210 mutation in the presence and in the absence of AZT. In AZT-naive patients, tryptophan at RT residue 210 was rare. After AZT exposure, W-210 appeared in a minority of those patients, most commonly in association with L-41 and Y-215. The presence of W-210 increased the AZTIC50 by two- to fourfold, as determined by both the recombinant virus assay and site-directed mutagenesis. A significant replication advantage in favor of the wild-type L-210 over W-210 was observed, although the selection against the 210 mutant was two- to threefold lower when the viruses were grown in the presence of 5 microM AZT. In summary, the L210W mutation appears to be of marginal significance, conferring approximately two- to fourfold-reduced sensitivity to AZT compared with similar AZT-resistant genomes with L-210. The selection pressure against W-210 may account for the modest proportion of patients in which W-210 appears in vivo. PMID- 8709215 TI - Full-length sequence and mosaic structure of a human immunodeficiency virus type 1 isolate from Thailand. AB - Human immunodeficiency virus type 1 isolates of envelope genotype E are contributing substantially to the global pandemic. These strains appear to be mosaics, with the gag gene from clade A and the envelope from clade E; the parental clade E strain has not been found. Here we report the first full genomic sequence of one such mosaic virus, isolate CM240 from Thailand. Multiple breakpoints between the two parental genotypes have been found in a CM240 virus. The entire gag-pol region and most, if not all, of the accessory genes vif, vpr, tat, rev, and vpu appear to derive from clade A. The genotype switches to E shortly after the signal peptide of the envelope and back to clade A near the middle of gp41; thus, the portion of the envelope that lies on the cytoplasmic side of the membrane appears to be principally derived not from clade E, as previously thought, but from clade A. Another small segment not belonging to any recognized clade and presumably also contributed by the parental E strain has been found in the long terminal repeat. It may be significant that the implied virion structure resembles a pseudotype virus with the matrix and core from one clade and the outer envelope from another. In the long terminal repeat, differences were observed between CM240 and other clades in the number of NF kappa B binding sites, the sequence of the TATA box, and the putative secondary structure of the transactivation response region stem-loop. The mosaic structure of a CM240 virion is suggestive of phenotypic differences which might have contributed to the emergence of this variant. PMID- 8709217 TI - Cleavage site mutations in the encephalomyocarditis virus P3 region lethally abrogate the normal processing cascade. AB - Site-specific mutations within the proteinase 3C-dependent P3 region cleavage sequences of encephalomyocarditis virus have been constructed. The mutations altered the normal QG cleavage site dipeptide pairs of the 2C/3A, 3A/3B, 3B/3C, and 3C/3D junctions into QV, QC, QF, QY, and RG sequences. When translated in vitro in the context of full-length viral polyproteins, all mutations blocked endogenous 3C-mediated processing at their engineered sites and produced stable forms of the expected viral P3 precursors that were also resistant to cleavage by exogenously added recombinant 3C. Relative to wild-type viral sequences, each mutant form of P3 had a somewhat different ability to mediate overall polyprotein processing. Mutations at the 2C/3A, 3A/3B, and 3B/3C sites, for example, were generally less impaired than 3C/3D mutations, when the cleavage reactions were quantitated with cotranslated L-P1-2A precursors. A notable exception was mutant 3B3C(QG-->RG), which proved far less active than sibling mutants 3B3C(QG-->QF) and 3B3C(QG-->QV), a finding that possibly implicates this segment in the proper folding of an active 3C. When transfected into HeLa cells, all mutant sequences were lethal, presumably because of the reduced L-P1-2A processing levels or reduced RNA synthesis capacity. However, when specifically tested for the latter activity, all mutations except those at the 3C/3D cleavage site were indeed able to initiate and perpetuate viral RNA replication in transfected cells, albeit to RNA accumulation levels lower than those produced by wild-type sequences. The transfection effects could be mimicked with cell-free synthesized proteins, in that translation samples containing locked 3CD polymerase precursors were catalytically inactive in poly(A)-oligo(U)-dependent assays, while all other mutant processing samples initiated detectable RNA synthesis. Surprisingly, not only did the 3B/3C mutant sequences prove capable of directing RNA synthesis, but the viral RNA thus synthesized could be immunolabeled and precipitated with 3C specific monoclonal antibody reagents, indicating an unexpected covalent attachment of the proteinase to the RNA product whenever this cleavage site was blocked. PMID- 8709216 TI - Transcriptional regulation of human polyomavirus JC: evidence for a functional interaction between RelA (p65) and the Y-box-binding protein, YB-1. AB - The transcriptional control region of the human neurotropic polyomavirus JC virus contains a consensus NF-kappa B site which has been shown to enhance both basal and extracellular stimulus-induced levels of transcription of JC promoters. Here, we show that the expression of JC late promoter constructs containing the NF kappa B site is decreased by cotransfection with the NF-kappa B/rel subunits, p50 and p52, but enhanced by the p65 subunit. However, JC promoter constructs lacking the NF-kappa B site were activated by p52 and p50 and repressed by p65. This antithetical response of the JC promoter mapped specifically to the D domain, which is a target site for the cellular transcription factor, YB-1. Band shift studies indicated that YB-1 and p65 modulate each other's binding to DNA: YB-1 augments the affinity of p65 for the NF-kappa B site, while p65 reduces the binding of YB-1 to the D domain. Results from coimmunoprecipitation followed by Western blot (immunoblot) analysis suggest an in vivo interaction between p65 and YB-1 in glial cells. Functionally, YB-1 appears to act synergistically with p65 to control transcription from the NF-kappa B site. A converse pattern is seen with the D domain, in which YB-1 acts synergistically with p50 and p52 to regulate transcription. p50 and p52 may function as transcriptional activators on the D domain by removing the repressive effect of p65 on YB-1 binding to the D domain. On the basis of these data, we propose a model in which NF-kappa B/rel subunits functionally interact with consensus NF-kappa B sites or YB-1-binding sites, with disparate effects on eukaryotic gene expression. PMID- 8709218 TI - Evidence that two latency-associated transcripts of herpes simplex virus type 1 are nonlinear. AB - The latency-associated transcripts (LATs) of herpes simplex virus type 1 (HSV-1) are the only viral gene products that accumulate to abundant levels in latently infected cells. Others have reported species of 2.0, 1.50, and 1.45 kb; only the 2.0-kb species is seen in productively infected cells, and there is evidence that it behaves as an intron. We examined the LATs both in trigeminal ganglia of latently infected mice and in productively infected cultures of monkey CV-1 cells. After glyoxalation, RNA was subjected to high-resolution agarose gel electrophoresis and Northern (RNA) analysis, a procedure capable of resolving linear and nonlinear RNA species. Under these conditions, we resolved the 2.0-kb LAT into two species; the slower species was much more abundant and had a mobility significantly slower than expected for a linear RNA. To test the hypothesis that this RNA was in fact nonlinear, we used partial hydrolysis by sodium carbonate and oligonucleotide-directed RNase H digestion. These procedures changed the mobility of the slower species into that of the faster species. Similarly, the mobility of the 1.50-kb LAT, which was much more abundant than the 1.45-kb LAT, was changed by these procedures to that of the 1.45-kb LAT. Our data show that the two major LAT species are nonlinear, and they support an interpretation of stable lariat structures. PMID- 8709219 TI - Effects of the route of infection on immunoglobulin G subclasses and specificity of the reovirus-specific humoral immune response. AB - Reovirus serotype 1, strain Lang (T1/L), a well characterized enteric virus, elicits humoral and cellular immune responses in mice. Although orally and intradermally induced infections generate comparable reovirus-specific serum antibody titers, little is known about the effects of the route of infection on the systemic immunoglobulin G (IgG) response. To assess whether the route of exposure affects virus-specific humoral immunity, we infected various strains of mice with reovirus T1/L by the oral or intradermal routes. At day 10 following infection, virus-specific serum antibody titers and IgG subclasses were determined by enzyme-linked immunosorbent assay. Serum IgG2a and IgG2b antibodies were detected in all mouse strains independent of the route of infection. Mice of the H-2d haplotype that received an intradermal infection also had high levels of reovirus-specific serum IgG1. This dichotomy of responses was not associated with differences in the types of cytokine produced by draining peripheral lymph nodes. However, peripheral lymph node lymphocytes from C3H mice produced significantly higher levels of gamma interferon than did BALB/c, C57BL/6, and B10.D2 mice. Additionally, peripheral lymph node lymphocytes from all strains of mice produced only low levels of interleukin-5, with no detectable level of interleukin-4 or interleukin-6. Analysis of specific antibody at inductive sites of the immune response showed that orally infected Peyer's patches produced predominantly IgA and intradermally infected peripheral lymph nodes produced predominantly IgG2a. Western blot (immunoblot) analysis showed that virus-specific IgA, IgG1, and IgG2a reacted with reovirus structural proteins. These data suggest that the route of infection affects the isotype and IgG subclasses, but not the antigen specificity, of the local antibody response. In addition, virus-specific IgG1 generated following an intradermally induced infection is linked to the H-2d major histocompatibility complex haplotype. PMID- 8709221 TI - Involvement of the mutated M protein in altered budding polarity of a pantropic mutant, F1-R, of Sendai virus. AB - Wild-type Sendai virus buds at the apical plasma membrane domain of polarized epithelial MDCK cells, whereas a pantropic mutant, F1-R, buds at both the apical and basolateral domains. In F1-R-infected cells, polarized protein transport and the microtubule network are impaired. It has been suggested that the mutated F and/or M proteins in F1-R are responsible for these changes (M. Tashiro, J. T. Seto, H.-D. Klenk, and R. Rott, J. Virol. 67:5902-5910, 1993). To clarify which gene or mutation(s) was responsible for the microtubule disruption which leads to altered budding of F1-R, MDCK cell lines containing the M gene of either the wild type or F1-R were established. When wild-type M protein was expressed at a level corresponding to that synthesized in virus-infected cells, cellular polarity and the integrity of the microtubules were affected to some extent. On the other hand, expression of the mutated F1-R M protein resulted in the formation of giant cells about 40 times larger than normal MDCK cells. Under these conditions, the effects on the microtubule network were enhanced. The microtubules were disrupted and polarized protein transport was impaired as indicated by the nonpolarized secretion of gp80, a host cell glycoprotein normally secreted from the apical domain, and bipolar budding of wild-type and F1-R Sendai viruses. The mutated F glycoprotein of F1-R was transported bipolarly in cells expressing the F1-R M protein, whereas it was transported predominantly to the apical domain when expressed alone or in cells coexpressing the wild-type M protein. These findings indicate that the M protein of F1-R is involved in the disruption of the microtubular network, leading to impairment of cellular polarity, bipolar transport of the F glycoprotein, and bipolar budding of the virus. PMID- 8709220 TI - Determination and analysis of the complete nucleotide sequence of human herpesvirus. AB - Human herpesvirus 7 (HHV-7) is a recently isolated betaherpesvirus that is prevalent in the human population, with primary infection usually occurring in early childhood. HHV-7 is related to human herpesvirus 6 (HHV-6) in terms of both biological and, from limited prior DNA sequence analysis, genetic criteria. However, extensive analysis of the HHV-7 genome has not been reported, and the precise phylogenetic relationship of HHV-7 to the other human betaherpesviruses HHV-6 and human cytomegalovirus has not been determined. Here I report on the determination and analysis of the complete DNA sequence of HHV-7 strain JI. The data establish that the close biological relationship of HHV-6 and HHV-7 is reflected at the genetic level, where there is a very high degree of conservation of genetic content and encoded amino acid sequences. The data also delineate loci of divergence between the HHV-6 and HHV-7 genomes, which occur at the genome terminal in the region of the terminal direct-repeat elements and within limited regions of the unique component. Of potential significance with respect to biological and evolutionary divergence of HHV-6 and HHV-7 are notable structural differences in putative transcriptional regulatory genes specified by the direct repeat and immediate-early region A loci of these viruses and the absence of an equivalent of the HHV-6 adeno-associated virus type 2 rep gene homolog in HHV-7. PMID- 8709222 TI - The posttranscriptional control element of the simian retrovirus type 1 forms an extensive RNA secondary structure necessary for its function. AB - It was previously shown that a 240-nucleotide (nt) RNA element (cis-acting transactivation element [CTE]) located between the env gene and the 3' long terminal repeat of simian retrovirus type 1 (SRV-1) can functionally replace posttranscriptional activation directed by Rev and the Rev-responsive element (RRE) when inserted into a Rev- and RRE-deficient molecular clone of human immunodeficiency virus type 1, resulting in efficient virus replication. Here, we analyze the molecular and structural requirements for function of this RNA element. Deletion mutagenesis demonstrated that the core element spans 173 nt. SRV-2 and Mason-Pfizer monkey virus have highly homologous elements, which function similarly when inserted into the Rev/RRE-deficient human immunodeficiency virus type 1. Computer prediction indicated that the core CTEs of all three viruses have similar extensive secondary structures. Mutagenesis of the SRV-1 CTE revealed that both sequence and secondary structure are essential for function. Nuclease probing of the SRV-1 CTE further supported the genetic analysis and confirmed the predicted structural features of the RNA element. Sequence analysis of the 240-nt SRV-1 CTE, after continuous long-term propagation of the Rev-independent viruses, revealed that the genetically defined core element remained unchanged, while regions outside the core CTE underwent deletions or duplications. These data further support our in vitro mutagenesis data and demonstrate the importance of the sequence and structure of the SRV-1 CTE for appropriate function. PMID- 8709223 TI - Regulation of the herpesvirus saimiri oncogene stpC, similar to that of T-cell activation genes, in growth-transformed human T lymphocytes. AB - Herpesvirus saimiri strain C488, a T-cell tumor virus of New World primates, transforms human T lymphocytes to stable interleukin-2-dependent growth without need for further stimulation by antigen or mitogen. The transformed cell lines show the phenotype of activated mature T cells and retain many essential features of the primary parental cells, e.g., antigen specificity. In contrast to transformed New World monkey T cells, the human lines do not support lytic growth of the virus, even after chemical stimulation. Here we show that many viral genes remain silent during episomal persistence. However, the viral oncogene stpC is predominantly transcribed and translated to a stable cytoplasmic protein of 20 kDa that is heterogeneously expressed in individual cells. This 1.7-kb mRNA is bicistronic, encoding also Tip, a viral protein interacting with the T-cell specific tyrosine kinase Lck. stpC/tip transcripts are heavily induced upon stimulation by mitogen or phorbol ester. Block of protein synthesis does not abolish transcription: treatment with cycloheximide greatly induces stpC/tip mRNA levels. Thus, this gene complex is regulated similarly to early T-cell activation genes. Constitutive and induced expression engage different transcription start sites. The T-cell regulation of the viral genes stpC and tip may contribute to the T-cell tropism of growth transformation by herpesvirus saimiri. PMID- 8709225 TI - Characterization of a 120-Kilodalton pre-S-binding protein as a candidate duck hepatitis B virus receptor. AB - Infection by human and animal hepadnaviruses displays remarkable host and tissue tropism. The infection cycle probably initiates with binding of the pre-S domain of viral envelope protein to surface receptors present on the hepatocyte. Three types of neutralizing monoclonal antibodies against duck hepatitis B virus (DHBV) have their binding sites clustered within residues 83 to 107 of the pre-S protein, suggesting that this region may constitute a major receptor binding site. A 170- or 180-kDa duck protein (p170 or gp180) which binds DHBV particles through this part of the pre-S sequence has been identified recently. Although the p170 binding protein is host (duck) specific, its distribution is not restricted to DHBV-infectible tissues. Using the pre-S protein fused to glutathione S-transferase and immobilized on Sepharose beads, we have now identified an additional binding protein with a size of 120 kDa (p120). p120 expression is restricted to the liver, kidney, and pancreas, the three major organs of DHBV replication. While optimal p170 binding requires an intact pre-S protein, binding to p120 occurs much more efficiently with a few N- or C terminally truncated forms. The p120 binding site was mapped to residues 98 to 102 of the pre-S region, which overlaps with a cluster of known virus neutralizing epitopes. Site-directed mutagenesis revealed residues 100 to 102 (Phe-Arg-Arg) as the critical p120 contact site; nonconservative substitution in any of the three positions abolished p120 binding. Double mutations at positions 100 to 102 markedly reduced DHBV infectivity in cell culture. Short pre-S peptides covering the clustered neutralizing epitopes (also p170 and p120 binding sites) reduced DHBV infectivity in primary duck hepatocyte cultures. Thus, p120 represents a candidate component of the DHBV receptor complex. PMID- 8709224 TI - Epstein-Barr virus nuclear protein 2 (EBNA2) binds to a component of the human SNF-SWI complex, hSNF5/Ini1. AB - Epstein-Barr nuclear antigen 2 (EBNA2), one of the six viral nuclear proteins expressed in latently infected B lymphocytes, is essential to the immortalization of B cells by Epstein-Barr virus (EBV). EBNA2 promotes transcriptional transactivation of viral and cellular genes by acting as an adapter molecule that binds to cellular sequence-specific DNA-binding proteins, JK recombination signal binding protein (RBP-JK), and PU.1 and engages multiple members of the RNA polymerase II transcription complex. In the present study, we show that EBNA2 also interacts with hSNF5/Ini1, the human homolog of the yeast transcription factor SNF5. Gel filtration fractionation of partially purified EBV-positive lymphocyte nuclear extracts shows that a fraction of EBNA2 coelutes with both hSNF5/Ini1 and BRG1, a human homolog of SWI/SNF2, in the high-molecular-mass region (1.5 to 2.0 MDa) of a Superose 6 chromatogram. An affinity-purified rabbit antibody directed against hSNF5/Ini1 coimmunoprecipitates EBNA2 from this high molecular-mass nuclear protein fraction, demonstrating that EBNA2 and hSNF5/Ini1 interact in vivo. This interaction is restricted to a subpopulation of phosphorylated viral EBNA2. Deletion mutation analysis of EBNA2 shows that the proline-rich aminoterminal end and a domain within the divergent region of EBNA2 mediate EBNA2-hSNF5/Ini1 interaction. Since the SNF-SWI complex participates in gene regulation through the alteration of nucleosome configuration and may be a component of the RNA polymerase II holoenzyme, the EBNA2-hSNF5/Ini1 interaction supports the hypothesis that EBNA2 facilitates transcriptional transactivation by acting as a transcription adapter molecule. We postulate that EBNA2 engages the hSNF-SWI complex to generate an open chromatin conformation at the EBNA2 responsive target genes, thereby potentiating the function of the RBP-JK-EBNA2 polymerase II transcription complex. PMID- 8709226 TI - Mutational analysis of the envelope protein of spleen necrosis virus. AB - Spleen necrosis virus (SNV) is an amphotropic type C retrovirus originally isolated from a duck. The envelope protein is related to that of type D retroviruses, and SNV appears to use the same receptor as do simian retroviruses. However, little is known about envelope-receptor interactions of SNV. We constructed a series of envelope mutants to characterize the SU peptide of SNV. Point mutations were introduced throughout SU in regions that are conserved among all retroviruses belonging to the same receptor interference group. The biological and biochemical properties of these mutants were analyzed. All mutants were transported efficiently to the cell surface. Almost all mutations in the amino-terminal one-third caused a conformational change of the envelope and a significant drop in infectivity and abolished the ability to confer superinfection interference. Similar observations were made with only two of seven mutants with mutations in the middle of SU. Four mutations in this region had little or no effect on biological activity. One mutant envelope protein (Asp to Arg at position 192) was processed normally but showed little infectivity and had no ability to confer superinfection interference. A detailed mutational analysis suggested that this amino acid forms a hydrogen bond to its cellular receptor. Mutations within the carboxy-terminal part of SU had very little or no effect on biological function. Aberrantly processed envelope proteins were proteolytically cleaved at a new point upstream of and differing in sequence from the conserved retroviral SU/TM cleavage site. Surprisingly, these mutants still retained some infectivity (0.01 to 1% of that of the wild type). Our data indicate that the envelope of SNV behaves in a manner very different from that of the envelopes of other studied retroviruses. PMID- 8709227 TI - CD4 down-modulation during infection of human T cells with human immunodeficiency virus type 1 involves independent activities of vpu, env, and nef. AB - The human immunodeficiency virus type 1 (HIV-1) genes vpu, env, and nef have all been implicated in modulating the levels of cell surface CD4 on infected cells. To quantitatively assess the relative contribution of each gene product to the regulation of CD4 during HIV infection of Jurkat T cells and peripheral blood mononuclear cells, we have developed an infectious HIV reporter system which expresses different combinations of these genes. To distinguish infected cells in the early or late stages of infection from uninfected cells, these viruses were designed to express human placental alkaline phosphatase with the kinetics of either early or late viral genes. Flow cytometry to detect placental alkaline phosphatase and CD4 in infected cells showed that vpu, env, and nef are independently capable of down-modulation of CD4. As predicted by their respective expression patterns, nef down-modulated CD4 rapidly during the early phase of virus infection whereas vpu and env functioned late in the infection. In both Jurkat cells and peripheral blood mononuclear cells, a combination of the three genes was more efficient than any one or two genes, demonstrating that all three genes are required to achieve maximal CD4 down-modulation. In primary cells, down modulation of CD4 was less efficient than in Jurkat cells and there was a stronger dependence on nef function for reducing cell surface CD4. HIV therefore has three genes that are able to independently down-modulate CD4; together, they can eliminate the bulk of cell surface CD4. PMID- 8709228 TI - Inducible gene expression by retrovirus-mediated transfer of a modified tetracycline-regulated system. AB - The ability to regulate gene expression via exogenous stimuli will facilitate the study of gene functions in mammalian cells. In the present study, we modified the tetracycline-controlled inducible system by the addition of the ligand-binding domain of the estrogen receptor to the carboxy terminus of the tTA transactivator. A single retroviral vector can transduce both the transactivator gene and the gene of interest controlled by the tTA-inducible promoter into mammalian cells. We show that cell lines expressing the transactivator can readily be established and that expression of the gene of interest depends on the removal of tetracycline and the addition of estrogen. By using this system, cell lines with inducible expression of the G protein of vesicular stomatitis virus, a potentially toxic gene product, were established. The combination of a powerful inducible system and retrovirus-mediated gene transfer can not only be used to study gene function but may also be applied in the future to clinical trials in human gene therapy. PMID- 8709229 TI - Interactions of the transcription factors MIBP1 and RFX1 with the EP element of the hepatitis B virus enhancer. AB - We previously demonstrated that MIBP1 and RFX1 polypeptides associate in vivo to form a complex that binds to the MIF-1 element in the c-myc gene and the major histocompatibility complex class II X-box recognition sequence. We now show that the EP element, a key regulatory sequence within hepatitis B virus enhancer I, also associates with MIBP1 and RFX1. Using polyclonal antisera directed against either oligonucleotide-purified MIBP1 or a peptide derived from the major histocompatibility complex class II promoter-binding protein RFX1, we showed that MIBP1 and RFX1 are both present in the DNA-protein complexes at the EP site. In addition, while the EP element can act cooperatively with several adjacent elements to transactivate hepatitis B virus expression, we demonstrated that the EP site alone can repress transcription of simian virus 40 promoter in a position and orientation-independent manner, suggesting a silencer function in hepatocarcinoma cells. PMID- 8709230 TI - Oligomeric structure of glycoproteins in herpes simplex virus type 1. AB - A number of herpes simplex virus (HSV) glycoproteins are found in oligomeric states: glycoprotein E (gE)-gI and gH-gL form heterodimers, and both gB and gC have been detected as homodimers. We have further explored the organization of glycoproteins in the virion envelope by using both purified virions to quantitate glycoprotein amounts and proportions and chemical cross-linkers to detect oligomers. We purified gB, gC, gD, and gH from cells infected with HSV type 1 and used these as immunological standards. Glycoproteins present in sucrose gradient purified preparations of two strains of HSV type 1, KOS and NS, were detected with antibodies to each of the purified proteins. From these data, glycoprotein molar ratios of 1:2:11:16 and 1:1:14:9 were calculated for gB/gC/gD/gH in KOS and NS, respectively. gL was also detected in virions, although we lacked a purified gL standard for quantitation. We then asked whether complexes of these glycoproteins could be identified, and if they existed as homo- or hetero oligomers. Purified KOS was incubated at 4 degrees C with bis (sulfosuccinimidyl) suberate (BS3), an 11.4 A (1A = 0.1 mm) noncleavable, water-soluble cross-linker. Virus extracts were examined by Western blotting (immunoblotting), or immunoprecipitation followed by Western blotting, to assay for homo- and hetero oligomers. Homodimers of gB, gC, and gD were detected, and hetero-oligomers containing gB cross-linked to gC, gC to gD, and gD to gB were also identified. gH and gL were detected as a hetero-oligomeric pair and could be cross-linked to gD or gC but not to gB. We conclude that these glycoproteins are capable of forming associations with one another. These studies suggest that glycoproteins are closely associated in virions and have the potential to function as oligomeric complexes. PMID- 8709231 TI - Cross-linking of glycoprotein oligomers during herpes simplex virus type 1 entry. AB - Herpes simplex virus (HSV) has 10 glycoproteins in its envelope. Glycoprotein B (gB), gC, gD, gH, and gL have been implicated in virus entry. We previously used chemical cross-linking to show that these five glycoproteins were close enough to each other to be cross-linked into homodimeric and hetero-oligomeric forms; hetero-oligomers of gB-gC, gC-gD, gD-gB, gH-gL, gC-gL and gD-gL were found in purified virions. To better understand the roles of these glycoproteins in viral entry, we have modified a standard HSV penetration assay to include cross linkers. This allowed us to examine changes in associations of viral glycoproteins during the entry process. HSV-1(KOS) was adsorbed at 4 degrees C to human neuroblastoma cells (SY5Y). The temperature was raised to 37 degrees C and cells were treated with cross-linker at various times after the temperature shift. Cytoplasmic extracts were examined by Western blotting (immunoblotting) for viral glycoproteins. We found that (i) as in virus alone, the length and concentration of the cross-linking agent affected the number of specific complexes isolated; (ii) the same glycoprotein patterns found in purified virions were also present after attachment of virions to cells; and (iii) the ability to cross-link HSV glycoproteins changed as virus penetration proceeded, e.g., gB and gD complexes which were present during attachment disappeared with increasing time, and their disappearance paralleled the kinetics of penetration. However, this phenomenon appeared to be selective since it was not observed with gC oligomers. In addition, we examined the cross-linking patterns of gB and gD in null viruses K082 and KOSgD beta. Neither of these mutants, which attach but cannot penetrate, showed changes in glycoprotein cross-linking over time. We speculate that these changes are due to conformational changes which preclude cross-linking or spatial alterations which dissociate the glycoprotein interactions during the penetration events. PMID- 8709232 TI - A reevaluation of the higher taxonomy of viruses based on RNA polymerases. AB - In order to assess the validity of classifications of RNA viruses, published alignments and phylogenies of RNA-dependent RNA and DNA polymerase sequences were reevaluated by a Monte Carlo randomization procedure, bootstrap resampling, and phylogenetic signal analysis. Although clear relationships between some viral taxa were identified, overall the sequence similarities and phylogenetic signals were insufficient to support many of the proposed evolutionary groupings of RNA viruses. Likewise, no support for the common ancestry of RNA-dependent RNA polymerases and reverse transcriptases was found. PMID- 8709233 TI - Proteins associated with purified human cytomegalovirus particles. AB - Virion-associated proteins isolated from purified human cytomegalovirus particles (strain AD169) were used as substrates for chemical sequence analysis. Extracellular virions, noninfectious enveloped particles, and dense bodies were purified by negative viscosity-positive density gradient centrifugation, and their component proteins were separated by denaturing polyacrylamide gel electrophoresis. The deduced amino acid sequence of individual protein bands was used to identify six corresponding viral genes whose products have not previously been identified as virion constituents: UL47, UL25, UL88, UL85, UL26, and UL48.5. In addition, a 45-kDa cellular protein was identified, and the protein fragments sequenced have a high degree of amino acid identity with actin. However, antiactin monoclonal and polyclonal antibodies did not react with a specific protein in the virus preparations, suggesting that this 45-kDa protein is an immunologically distinct isoform of actin. The newly identified viral and cellular proteins were resistant to protease treatment of purified virions, suggesting that they are unlikely to be contaminants of the viral preparations. PMID- 8709234 TI - Characterization of human immunodeficiency virus type 1 Vif particle incorporation. AB - The human immunodeficiency virus type 1 (HIV-1) Vif protein is necessary at the time of viral particle formation yet functionally manifests its effect after virions enter target cells. This suggests that Vif either acts on another viral protein or is itself incorporated into particles. In this study, we have examined the latter possibility. We confirm our previous observation that Vif is incorporated into human immunodeficiency virus type 1 virions at a ratio of approximately 1 molecule of Vif for every 75 to 220 molecules of p24, or 7 to 20 molecules per virion. Furthermore, we demonstrate that the relative concentration of Vif is much lower in particles than in infected cells, whereas the opposite is observed for the main virus components. The viral envelope, Nef, Vpr, Vpu, protease, reverse transcriptase, integrase, nucleocapsid, and p6gag proteins as well as the viral genomic RNA are dispensable for Vif packaging. Furthermore, mutating several highly conserved residues (H-108, C-114, C-133, L-145, and Q 146) or deleting the C-terminal 18 amino acids of Vif, either of which severely impairs Vif function, does not abolish its incorporation into virions. Finally, Vif can be packaged into murine leukemia virus particles. On the basis of these data, we conclude that the specificity of Vif incorporation into virions remains an open question. PMID- 8709235 TI - Functional interaction of paramyxovirus glycoproteins: identification of a domain in Sendai virus HN which promotes cell fusion. AB - The cell fusion activity of most paramyxoviruses requires coexpression of a fusion protein (F) and a hemagglutinin-neuraminidase protein (HN) which are derived from the same virus type. To define the domain of the HN protein which interacts with the F protein in a type-specific manner a series of chimeric HN proteins between two different paramyxoviruses, Sendai virus (SN) and human parainfluenza virus type 3 (PI3), was constructed and coexpressed with the SN-F protein by using the vaccinia virus T7 RNA polymerase transient-expression system. Quantitative assays were used to evaluate cell surface expression as well as fusion-promoting activities of the chimeric HN molecules. A chimeric HN protein [SN(140)] containing 140 N-terminal amino acids derived from SN-HN and the remainder (432 amino acids) derived from PI3-HN was found to promote cell fusion with the SN-F protein. In contrast, a second chimeric HN with 137 amino acids from SN-HN at the N terminus could not promote fusion with SN-F, even though the protein was expressed on the cell surface. A construct in which the PI3-HN cytoplasmic tail and transmembrane domain were substituted for those of SN in the SN(140) chimera still maintained the ability to promote cell fusion. These results indicate that a region including only 82 amino acids in the extracellular domain, adjacent to the transmembrane domain of the SN-HN protein, is important for interaction with the SN-F protein and promotion of cell fusion. PMID- 8709236 TI - Influenza virus nucleoprotein-specific immunoglobulin G subclass and cytokine responses elicited by DNA vaccination are dependent on the route of vector DNA delivery. AB - Endpoint immunoglobulin G (IgG) titers and cytotoxic T-lymphocyte (CTL) activities were identical between mice immunized via the intramuscular and epidermal (gene gun) routes with 100 and 1 micrograms, respectively, of an influenza virus nucleoprotein (NP) expression vector. However, examination of the relative levels of two IgG subclasses demonstrated that muscle inoculation resulted in predominantly IgG2a responses, whereas gene gun immunization yielded a preponderance of IgG1 antibodies. Inasmuch as these data suggested that muscle inoculation and gene gun delivery elicited Th1-like and Th2-like responses, respectively, gamma interferon release profiles from antigen-stimulated splenocytes were remarkably similar between these groups. Interleukin-4 (IL-4) production assays, on the other hand, revealed qualitative differences that could be correlated with the divergent IgG subclass data. Waning gamma interferon production in gene gun-immunized animals was countered by a marked increase in IL 4 production following the third immunization, as was the case in control animals immunized with inactivated influenza virus formulated with Freund's adjuvant. In contrast, significant levels of IL-4 production were not observed in the intramuscular DNA inoculation group, despite similar decreases in gamma interferon production with increasing immunizations. These data show that intramuscular inoculation leads to Th1-like responses due to elevated IgG2a levels, production of gamma interferon, CTL activity, and lack of IL-4. However, gene gun responses are more difficult to categorize because of the presence of significant gamma interferon and CTL activity on the one hand and elevated IgG1 antibodies and increasing IL-4 production with successive immunizations on the other. In addition, there was a lack of correlation between IgG isotype ratios and cytokine production in all of the NP DNA-immunized animals, in that IgG subclass ratios remained fixed while cytokine production patterns fluctuated with successive immunizations. These data are consistent with the idea that the types of responses elicited following DNA immunization. are dependent on both the identity of the antigen and the route of DNA administration. PMID- 8709237 TI - Translation initiation in GB viruses A and C: evidence for internal ribosome entry and implications for genome organization. AB - GB viruses A and C (GBV-A and GBV-C) are two recently described RNA viruses which appear to be members of the Flaviviridae. Although both viruses appear to contain long 5' nontranslated regions, the sites of polyprotein initiation and the presence of core-like proteins remain to be determined. Translation studies were undertaken to determine the mechanism and sites of polyprotein initiation in GBV A and GBV-C. Rabbit reticulocyte lysates programmed with monocistronic RNAs containing 5' ends of GBV-A or GBV-C fused in-frame with the chloramphenicol acetyltransferase (CAT) open reading frame generated GBV-CAT fusion proteins in vitro. Site-specific mutagenesis and N-terminal sequencing located the sites of translation initiation immediately upstream of the putative signal sequence for the GBV E1 envelope glycoproteins. Efficient translation of the monocistronic GBV CAT RNAs required the inclusion of GBV coding sequences. This, coupled with the presence of at least 523 nucleotides of 5' nontranslated RNA containing multiple AUG codons, suggests that translation initiation of these RNAs did not utilize a ribosome scanning mechanism. Translation of bicistronic RNAs containing 5' nontranslated sequences within the intercistronic space was consistent with the presence of a weakly active internal ribosome entry site in both GBV-A and GBV-C. Secondary structure predictions indicate that the 5' ends of these viruses assume similar complex structures distinct from those identified in the internal ribosome entry site-containing picornaviruses, pestiviruses, and hepatitis C viruses. The data indicate that GBV-A and GBV-C are unique members of the Flaviviridae that do not contain core-like proteins at the N termini of their putative polyproteins. PMID- 8709238 TI - Increased envelope spike density and stability are not required for the neutralization resistance of primary human immunodeficiency viruses. AB - Previous observations that the gp120 envelope glycoprotein contents of some primary, clade B human immunodeficiency virus type 1 (HIV-1) isolates were higher than those of laboratory-passaged HIV-1 isolates suggested the hypothesis that increased envelope glycoprotein spike density or stability contributes to the relative neutralization resistance of the primary viruses. To test this, the structural, replicative, and neutralization properties of a panel of recombinant viruses with HIV-1 envelope glycoproteins from divergent clades were examined in an env complementation assay. In this system, although the spike density and stability of envelope glycoproteins from primary HIV-1 isolates were not greater than those from a laboratory-adapted isolate, relative resistance to neutralizing antibodies and soluble CD4 was observed for the viruses with primary envelope glycoproteins. Thus, neither high envelope glycoprotein spike density nor stability is necessary for the relative neutralization resistance of primary HIV 1 viruses. PMID- 8709239 TI - The structurally diverse intergenic regions of respiratory syncytial virus do not modulate sequential transcription by a dicistronic minigenome. AB - The first nine genes of respiratory syncytial virus (RSV), a nonsegmented negative-strand RNA virus, are separated by intergenic regions which range in size from 1 to 52 nucleotides for strain A2 and lack obvious consensus elements except that each ends in an A (genome sense). Their significance for gene expression was investigated by using RSV-CAT-LUC RNA, a helper-dependent cDNA encoded dicistronic analog of RSV genomic RNA in which the viral genes were replaced by a negative-sense copy of the translational open reading frame (ORF) encoding chloramphenicol acetyltransferase (CAT) as the upstream, leader-proximal gene and that encoding luciferase (LUC) as the downstream gene. These foreign ORFs were flanked by the RSV gene-start (GS) and gene-end (GE) transcription signals and separated by the naturally occurring G/F intergenic region. The RSV CAT-LUC minigenome was synthesized in vitro and transfected into RSV-infected cells, and synthesis of the CAT and LUC mRNAs was monitored by enzyme assay and Northern (RNA) blot hybridization. Surprisingly, substitution of each of the other naturally occurring RSV intergenic regions in turn did not significantly alter the absolute or relative amounts of the two mRNAs. Substitution of a nonnatural 10-nucleotide intergenic region, or elimination of the intergenic region altogether, also had little effect on the level of expression of the two genes. Four of the minigenome variants containing naturally occurring intergenic regions were modified further by replacing part of the LUC ORF with a second copy of the CAT ORF, so that each of the two mRNAs would hybridize equally with a CAT specific probe and their relative molar amounts could be determined. The level of expression of the downstream gene was 0.30 to 0.36 that of the upstream one. This determined the magnitude of RSV transcriptional polarity across a gene pair and confirmed that this value was very similar among the various intergenic regions. Minigenome transcription also yielded a CAT-LUC readthrough mRNA at a level 0.10 to 0.13 that of the LUC mRNA. In summary, the structurally diverse RSV intergenic regions do not appear to play a role in modulating RSV gene expression. PMID- 8709240 TI - Amino acids essential for RNase H activity of hepadnaviruses are also required for efficient elongation of minus-strand viral DNA. AB - The hepadnavirus P gene contains amino acid sequences which share homology with all known RNases H. In this study, we made four mutants in which single amino acids of the duck hepatitis B virus (DHBV) RNase H region were altered. In two of them, amino acids at locations comprising the putative catalytic site were changed, while the remaining mutants had alterations at amino acids conserved among hepadnaviruses. Transfection of these mutant genomes into permissive cells resulted in synthesis of several discrete viral nucleic acid species, ranging in apparent sizes from approximately 500 to 3,000 bp, numbered I, II, III, IV, and V. While the locations of the species were similar in all mutants, the proportions of the species varied among the mutants. Analysis of the nucleic acid species revealed that they were hybrid molecules of RNA and minus-strand DNA, indicating that the RNase H activity was missing or greatly reduced in these mutants. Primer extension experiments showed that the mutant viruses initiated minus-strand viral DNA synthesis normally. The 3' termini of minus-strand DNA in species II, III, and IV were mapped just downstream of nucleotides 1659, 1220, and 721, respectively. Species V contained essentially full-length minus-strand viral DNA. A parallel amino acid change in the putative catalytic site of the HBV RNase H domain resulted in accumulation of low-molecular-weight hybrid molecules consisting of RNA and minus-strand DNA and similar in size and pattern to those seen with DHBV. These studies demonstrate experimentally the involvement of the C terminal portion of the P gene in RNase H activity in both DHBV and human hepatitis B virus and indicate that the amino acids essential for RNase H activity of hepadnavirus P protein are also important for the efficient elongation of minus-strand viral DNA. PMID- 8709242 TI - Mutational analysis of the RNA triphosphatase component of vaccinia virus mRNA capping enzyme. AB - Vaccinia virus mRNA capping enzyme is a multifunctional protein with RNA triphosphatase, RNA guanylyltransferase, and RNA (guanine-7-) methyltransferase activities. The enzyme is a heterodimer of 95- and 33-kDa subunits encoded by the vaccinia virus D1 and D12 genes, respectively. The N-terminal 60-kDa of the D1 subunit (from residues 1 to 545) is an autonomous domain which catalyzes the triphosphatase and guanylyltransferase reactions. Mutations in the D1 subunit that specifically inactivate the guanylyltransferase without affecting the triphosphatase component have been described (P. Cong and S. Shuman, Mol. Cell. Biol. 15:6222-6231, 1995). In the present study, we identified two alanine cluster mutations of D1(1-545), R77A-K79A and E192A-E194A, that selectively inactivated the triphosphatase, but not the guanylyltransferase. Concordant mutational inactivation of RNA triphosphatase and nucleoside triphosphatase functions (to approximately 1% of wild-type specific activity) suggests that both gamma-phosphate cleavage reactions occur at a single active site. The R77A-K79A and E192A-E194A mutant enzymes were less active than wild-type D1(1-545) in the capping of triphosphate-terminated poly(A) but could be complemented in vitro by D1(1-545)-K260A, which is inert in nucleotidyl transfer but active in gamma phosphate cleavage. Whereas wild-type D1(1-545) formed only the standard GpppA cap, the R77A-K79A and E192A-E194A enzymes synthesized an additional dinucleotide, GppppA. This finding illuminates a novel property of the vaccinia virus capping enzyme, the use of triphosphate RNA ends as an acceptor for nucleotidyl transfer when gamma-phosphate cleavage is rate limiting. PMID- 8709241 TI - Human immunodeficiency virus type 1 Nef associates with a member of the p21 activated kinase family. AB - Although human immunodeficiency virus (HIV) Nef is essential for the induction of AIDS, its biochemical function has remained an enigma. In this study, HIV Nef protein is shown to associate with a serine-threonine kinase that recognizes histone H4 as a substrate, is serologically related to rat p21-activated kinase (PAK), and is specifically activated by Rac and Cdc42. These characteristics define the Nef-associated kinase as belonging to the PAK family. PAKs initiate kinase cascades in response to environmental stimuli, and their identification as a target of Nef implicates these signaling molecules in HIV pathogenesis and provides a novel target for clinical intervention. PMID- 8709243 TI - Transcriptional and replicational activation functions in the bovine papillomavirus type 1 E2 protein are encoded by different structural determinants. AB - A set of E2 proteins with mutations in the amino-terminal transactivation domain was made by a scheme called clustered charged-to-alanine scan. These mutant E2 proteins were tested for expression, stability, and compartmentalization in cells and for sequence-specific DNA binding, as well as in functional assays for transcriptional and replicational activation. We identified four groups of mutants. First, mutants K111A, K112A, and E176A were unable to activate replication and transcription because of oligomerization-induced retention of oligomers in the cytoplasm. Second, although fractions of the mutant proteins E74A and D143A/ R172C existed in the oligomeric form, they were localized in the nucleus. Certain fractions of these proteins existed as a dimer able to form a specific complex and activate replication; however, these proteins were inactive in transcriptional activation. Third, mutants R37A and D122A were localized in the nucleus, existed in the dimeric form, supported replication efficiently, and were severely crippled in transcriptional activation. The fourth group of mutants did not differ considerably from the wild-type protein. The activation of transcription by the wild type as well as mutant E2 proteins was dependent on the concentration of input E2 expression vector DNA and had a bell-like shape. We suggest that the reduction of transcriptional activation at higher E2 concentrations, the self-squelching activity, is caused by oligomerization of the E2 transactivator and is one of the mechanisms for the regulation of E2 activity. Our results also show that transcriptional and replicational activation activities are encoded by different determinants in the E2 protein. PMID- 8709244 TI - Inhibitors of human immunodeficiency virus type 1 zinc fingers prevent normal processing of gag precursors and result in the release of noninfectious virus particles. AB - The Cys-Xaa2-Cys-Xaa4-His-Xaa4-Cys zinc fingers of retroviral nucleocapsid proteins are prime antiviral targets because of conservation of the Cys and His chelating residues and the absolute requirement of these fingers in both early and late phases of retroviral replication. We previously reported that certain disulfide-substituted benzamides (DIBAs) chemically modify the Cys residues of the fingers, resulting in inhibition of human immunodeficiency virus type 1 (HIV 1) replication (W. G. Rice, J. G. Supko, L. Malspeis, R. W. Buckheit, Jr., D. Clanton, M. Bu, L. Graham, C. A. Schaeffer, J. A. Turpin, J. Domagala, R. Gogliotti, J. P. Bader, S. M. Halliday, L. Coren, R. C. Sowder II, L. O. Arthur, and L. E. Henderson, Science 270:1194-1197, 1995). We now examine the consequences of the interaction of DIBAs with the zinc fingers of the HIV-1 p7 nucleocapsid protein and its Pr55gag precursor. In HIV-1-infected U1 cells, DIBAs inhibited the release of infectious virions, and even under conditions in which virion particles were produced, the particles were noninfectious. DIBAs caused abnormal processing of Gag precursors, and the inhibitory effect on processing was not due to inhibition of the HIV-1 protease enzyme or Pr55gag myristoylation. Rather, the defect in processing was due to the formation of intermolecular cross linkages among the zinc fingers of adjacent Gag molecules, rendering the precursors no longer recognizable by HIV-1 protease. Likewise, DIBAs caused intermolecular cross-linkage among recombinant Pr55gag packaged into pseudovirions, thereby generating modified precursors that were resistant to the action of protease. Thus, DIBAs chemically modified the mutationally intolerant retroviral zinc fingers in infected cells, interrupting protease-mediated maturation of virions and leading ultimately to the production of compromised virions. PMID- 8709245 TI - Casein kinase II and protein kinase C modulate hepatitis delta virus RNA replication but not empty viral particle assembly. AB - Hepatitis delta virus (HDV) contains two virus-specific delta antigens (HDAgs), large and small forms, which are identical in sequence except that the large one contains 19 extra amino acids at the C terminus. HDAgs are nuclear phosphoproteins with distinct biological functions; the small form activates HDV RNA replication, whereas the large form suppresses this process but is required for viral particle assembly. In this study, we have characterized the phosphorylative property of HDAg in a human hepatoma cell line (HuH-7) and examined the role of phosphorylation in HDAg function. As demonstrated by in vivo labeling and kinase inhibitor experiments, the phosphorylation levels of both HDAgs were diminished by the inhibitor of casein kinase II (CKII). Nevertheless, phosphorylation of only the small form could be markedly reduced by the protein kinase C (PKC) inhibitor, suggesting different phosphorylation properties between the two HDAgs. When these two kinase inhibitors were added separately to the transient-expression system, HDV RNA replication was profoundly suppressed. In contrast, the inhibitors did not affect the assembly of empty HDAg particle from HDAgs and hepatitis B virus surface antigen. To further examine the role of phosphorylation in HDAg function, two conservative CKII recognition sites at Ser 2 and Ser-123 of both HDAgs and one potential PKC recognition site at Ser-210 of the large HDAg were altered to alanine by site-directed mutagenesis. Transfection experiments indicated that mutation at Ser-2, but not Ser-123, significantly impaired the activity of the small HDAg in assisting HDV RNA replication. This property is in accordance with our observation that Ser-2, not Ser-123, was the predominant CKII phosphorylation site in the small HDAg. Our studies also excluded the possibility that the phosphorylation of Ser-2, Ser-123, or Ser-210, had roles in the trans-suppression activity of the large HDAg, in the assembly of empty virus-like HDAg particle, and in the nuclear transport of HDAgs. In conclusion, our results indicate that both CKII and PKC positively modulate HDV RNA replication but not the assembly of empty HDAg particle. The role of CKII in HDV replication may at least in part be accounted for by the phosphorylation of Ser-2 in the small HDAg. The effect of PKC on HDV RNA replication is, however, not to mediate the phosphorylation of the conservative Ser-210 in the large HDAg but rather to act on as-yet-unidentified Ser or Thr residues in the small HDAg or cellular factors. These findings provide the first insight into the roles of phosphorylation of the two HDAgs in the HDV replication cycle. PMID- 8709246 TI - A longitudinal study of feline immunodeficiency virus-specific cytotoxic T lymphocytes in experimentally infected cats, using antigen-specific induction. AB - The evolution of the virus-specific cytotoxic T-lymphocyte response in two cats experimentally infected with feline immunodeficiency virus (FIV) was monitored. Effector cells were derived from peripheral blood lymphocytes during the acute and chronic phases of infection (0 to 21 and 62 to 127 weeks, respectively) and from the spleen and lymph nodes at 127 weeks after infection. Lymphocytes were restimulated in vitro with paraformaldehyde-fixed, autologous lymphoblasts which had been infected with recombinant vaccinia viruses expressing FIV GAG or ENV proteins. Unstimulated lymphocytes were also used as effectors in some assays. 51Cr-labelled autologous skin fibroblasts infected with recombinant vaccinia viruses were used as targets. FIV GAG-specific cytotoxic precursors were detected in restimulated circulating lymphocytes during acute infection in both cats. The onset of this activity was as early as 2 weeks postinfection (p.i.) in one cat. From 62 weeks p.i. neither FIV GAG- nor ENV-specific precursors could be detected in the peripheral blood. However, at 127 weeks p.i., GAG- and ENV-specific cytotoxic precursors were detected in lymphocytes isolated from lymph nodes. The FIV-specific cytotoxic cells were predominantly major histocompatibility complex class I restricted. No cytotoxic activity was detected from unstimulated lymphocytes. These studies demonstrate the use of an assay system for dissecting the FIV-specific cytotoxic cell response and show that precursor cells appear in the circulation very early after infection and prior to a detectable antibody response. Our results also suggest that the persistent high-level circulating antiviral cytotoxic T-lymphocyte responses seen in human immunodeficiency virus infected humans may not be a feature of FIV infections in cats. PMID- 8709247 TI - Adenovirus type 5 early region 4 is responsible for E1A-induced p53-independent apoptosis. AB - In the absence of E1B, the 289- and 243-residue E1A products of human adenovirus type 5 induce p53-dependent apoptosis. However, our group has shown recently that the 289-residue E1A protein is also able to induce apoptosis by a p53-independent mechanism (J. G. Teodoro, G. C. Shore, and P. E. Branton, Oncogene 11:467-474, 1995). Preliminary results suggested that p53-independent cell death required expression of one or more additional adenovirus early gene products. Here we show that both the E1B 19-kDa protein and cellular Bcl-2 inhibit or significantly delay p53-independent apoptosis. Neither early region E2 or E3 appeared to be necessary for such cell death. Analysis of a series of E1A mutants indicated that mutations in the transactivation domain and other regions of E1A correlated with E1A-mediated transactivation of E4 gene expression. Furthermore, p53-deficient human SAOS-2 cells infected with a mutant which expresses E1B but none of the E4 gene products remained viable for considerably longer times than those infected with wild-type adenovirus type 5. In addition, an adenovirus vector lacking both E1 and E4 was unable to induce DNA degradation and cell killing in E1A-expressing cell lines. These data showed that an E4 product is essential for E1A-induced p53 independent apoptosis. PMID- 8709248 TI - Identification of latent membrane protein 2A (LMP2A) domains essential for the LMP2A dominant-negative effect on B-lymphocyte surface immunoglobulin signal transduction. AB - Epstein-Barr virus (EBV) recombinants which carry three different deletion mutations in the LMP2A cytoplasmic amino-terminal domain were constructed. The presence of each mutation, LMP2A delta 21-36, LMP2A delta 21-64, and LMP2A delta 21-85, in EBV-infected transformed lymphoblastoid cell lines was confirmed by PCR analysis and Southern blot hybridization. Confirmation of mutant LMP2A protein expression was by immunofluorescence and immunoblotting with a newly identified rat monoclonal antibody that recognizes each of the LMP2A deletion mutations. Lymphoblastoid cell lines infected with recombinant EBV DNAs containing the mutations were analyzed for loss of LMP2A's dominant-negative effect on surface immunoglobulin signal transduction by monitoring induction of tyrosine phosphorylation, calcium mobilization, and activation of lytic replication following surface immunoglobulin cross-linking. Domains of LMP2A important for induction of tyrosine phosphorylation, calcium mobilization, and activation of lytic replication were identified. PMID- 8709249 TI - Complete replication in vitro of tobacco mosaic virus RNA by a template dependent, membrane-bound RNA polymerase. AB - A crude membrane-bound RNA polymerase, obtained by differential centrifugation of extracts of tomato leaves infected with tobacco mosaic tobamovirus (tomato strain L) TMV-L), was purified by sucrose density gradient centrifugation. Removal of the endogenous RNA template with micrococcal nuclease rendered the polymerase template dependent and template specific. The polymerase was primer independent and able to initiate RNA synthesis on templates containing the 3'-terminal sequences of the TMV-L positive or negative strands. TMV-vulgare RNA was a less efficient template, while RNAs of cucumber mosaic cucumovirus and red clover necrotic mosaic dianthovirus, or 5'-terminal sequences of TMV-L positive or negative strands, did not act as templates for the polymerase. A main product of the reaction with TMV-L genomic RNA as a template, carried out in the presence of [alpha-32P]UTP, was genomic-length single-stranded RNA. This was shown to be the positive strand and uniformly labelled along its length, demonstrating complete replication of TMV-L RNA. Genomic-length double-stranded RNA, labelled in both strands, and small amounts of RNAs corresponding to the single- and double stranded forms of the coat protein subgenomic mRNA were also formed. Antibodies to N-terminal and C-terminal portions of the 126-kDa protein detected the 126-kDa protein and the 183-kDa readthrough protein in purified RNA polymerase preparations, whereas antibodies to the readthrough portion of the 183-kDa protein detected only the 183-kDa protein. All three antibodies inhibited the template-dependent RNA polymerase, but none of them had any effect on the template-bound enzyme. PMID- 8709250 TI - Multivariate analysis of human immunodeficiency virus type 1 neutralization data. AB - We report on the use of spectral map analysis of the inter- and intraclade neutralization data of 14 sera of human immunodeficiency virus type 1 (HIV-1) infected individuals and 16 primary isolates, representing genetic clades A to H in group M and group O. This multivariate analysis has been used previously to study the interaction between drugs and receptors and between viruses and antiviral compounds. The analysis reveals the existence of neutralization clusters, not correlated with the known genetic clades. The structural factors that have been identified may correlate with the most important neutralization epitopes. Three key primary HIV-1 isolates, which allow discrimination of sera that are likely or unlikely to neutralize primary isolates from most of the genetic clades, were identified. Our method of analysis will facilitate the evaluation as well as the design of suitable HIV-1 vaccines, which induce high titer interclade cross-neutralizing antibodies. PMID- 8709251 TI - Abortive infection of the baculovirus Autographa californica nuclear polyhedrosis virus in Sf-9 cells after mutation of the putative DNA helicase gene. AB - Homologous recombination between the Autographa californica nuclear polyhedrosis virus (AcNPV) genome and a 0.6-kbp-long DNA fragment derived from the putative DNA helicase gene of Bombyx mori nuclear polyhedrosis virus generates eh2-AcNPV, an expanded-host-range AcNPV mutant (S. Maeda, S.G. Kamita, and A. Kondo, J. Virol. 67:6234-6238, 1993). After inoculation at a high multiplicity of infection (MOI), eh2-AcNPV replicates efficiently in both the Sf-9 (AcNPV-permissive) and BmN (non-AcNPV-permissive) cell lines. In this study, we found that after the inoculation of Sf-9 cells at a low MOI (i.e., 1 and 0.1 PFU per cell), the release of eh2-AcNPV virions was dramatically reduced (approximately 900- and 10,000-fold, respectively, at 72 h postinoculation) compared with that of wild type AcNPV. In addition, the titer of eh2-AcNPV determined by plaque assay on Sf 9 cells was approximately 200-fold lower than that determined by plaque assay on BmN cells. Analyses of gene expression and viral DNA replication after low-MOI eh2-AcNPV inoculation of Sf-9 cells indicated that viral early genes were expressed normally. However, DNA replication and late-gene expression were significantly reduced. These findings suggested that abortive infection occurred at the stage of viral DNA replication in nearly all low-MOI eh2-AcNPV-infected Sf 9 cells. In the larvae of Spodoptera frugiperda, the organism from which Sf-9 cells are derived, the infectivity of eh2-AcNPV was lower than that of AcNPV; however, abortive infection was not found. PMID- 8709252 TI - Apoptotic suppression by baculovirus P35 involves cleavage by and inhibition of a virus-induced CED-3/ICE-like protease. AB - Baculovirus p35 prevents programmed cell death in diverse organisms and encodes a protein inhibitor (P35) of the CED-3/interleukin-1 beta-converting enzyme (ICE) related proteases. By using site-directed mutagenesis, we have identified P35 domains necessary for suppression of virus-induced apoptosis in insect cells, the context in which P35 evolved. During infection, P35 was cleaved within an essential domain at or near the site DQMD-87G required for cleavage by CED-3/ICE family proteases. Cleavage site substitution of alanine for aspartic acid at position 87 (D87A) of the P1 residue abolished P35 cleavage and antiapoptotic activity. Although the P4 residue substitution D84A also caused loss of apoptotic suppression, it did not eliminate cleavage and suggested that P35 cleavage is not sufficient for antiapoptotic activity. Apoptotic insect cells contained a CED 3/ICE-like activity that cleaved in vitro-translated P35 and was inhibited by recombinant wild-type P35 but not P1- or P4-mutated P35. Thus, baculovirus infection directly or indirectly activates a novel CED-3/ICE-like protease that is inhibited by P35, thereby preventing virus-induced apoptosis. Our findings confirmed the inhibitory activity of P35 towards the CED-3/ICE protease, including recombinant mammalian enzymes, and were consistent with a mechanism involving P35 stoichiometric interaction and cleavage. P35's inhibition of phylogenetically diverse proteases accounts for its general effectiveness as an apoptotic suppressor. PMID- 8709253 TI - Insulin-like growth factor II blocks apoptosis of N-myc2-expressing woodchuck liver epithelial cells. AB - N-myc2 and insulin-like growth factor II (IGF-II) are coordinately overexpressed in the great majority of altered hepatic foci, which are the earliest precancerous lesions observed in the liver of woodchuck hepatitis virus carrier woodchucks, and these genes continue to be overexpressed in hepatocellular carcinomas (HCCs). We have investigated the function of these genes in woodchuck hepatocarcinogenesis by using a woodchuck liver epithelial cell line (WC-3). WC-3 cells react positively with a monoclonal antibody (12.8.5) against woodchuck oval cells, suggesting a lineage relationship with oval cells. Overexpression of N myc2 in three WC-3 cell lines caused their morphological transformation and increased their growth rate and saturation density in medium containing 10% serum. Removal of serum from the medium increased cell death of the N-myc2 expressing lines, whereas cell death in control lines was minimal. The death of N myc2-expressing WC-3 cells was accompanied by nucleosomal fragmentation of cellular DNA, and DAPI (4',6-diamidino-2-phenylindole) staining revealed condensation and fragmentation of the nuclei, suggesting that N-myc2-expressing WC-3 cells undergo apoptosis in the absence of serum. In colony regression assays, conducted in the absence of serum, control colonies were stable, while N myc2-expressing colonies regressed to various degrees. Addition of recombinant human IGF-II to the serum-free medium blocked both cell death and colony regression in all the N-myc2-expressing lines. Therefore, coordinate overexpression of N-myc2 and IGF-II in woodchuck altered hepatic foci may allow cells which otherwise might die to survive and progress to hepatocellular carcinoma. PMID- 8709254 TI - Autoantigens interact with cis-acting elements of rubella virus RNA. AB - Rubella virus (RV) infections in adult women can be associated with acute and chronic arthritic symptoms. In many autoimmune individuals, antibodies are found targeting endogenous proteins, called autoantigens, contained in ribonucleoprotein complexes (RNPs). In order to understand the molecular mechanisms involved in the RV-associated pathology, we investigated the nature of cellular factors binding RV RNA and whether such RNPs were recognized by antibodies in infected individuals. Previously, we noted that cellular proteins associated with the RV 5'(+) stem-loop (SL) RNA are recognized by serum with Ro reactivity. To better understand the nature of the autoantigens binding RV cis acting elements, serum samples from individuals with various autoimmune diseases were tested for their ability to immunoprecipitate RNPs containing labeled RV RNAs. A subset of serum samples recognizing autoantigen La, or Ro and La, immunoprecipitated both the RV 5'(+)SL and 3'(+)SL RNA-protein complexes. Autoantigens binding the RV 5'(+)SL and 3'(+)SL RNAs differed in molecular mass, specificities for respective RNA binding substrates, and sensitivity to alkaline phosphatase treatment. The La autoantigen was found to interact with the RV 5'(+)SL RNA as determined by immunological techniques and binding reactions with mixtures containing recombinant La protein. To test whether there is a correlation between La binding to an RV RNA element and the appearance of an anti La response, we measured anti-La titers in RV-infected individuals. Significant anti-La activity was detected in approximately one-third of RV-infected individuals 2 years postinfection. PMID- 8709255 TI - Cell proteins bind specifically to West Nile virus minus-strand 3' stem-loop RNA. AB - The first 96 nucleotides of the 5'noncoding region (NCR) of West Nile virus (WNV) genomic RNA were previously reported to form thermodynamically predicted stem loop (SL) structures that are conserved among flaviviruses. The complementary minus-strand 3' NCR RNA, which is thought to function as a promoter for the synthesis of plus-strand RNA, forms a corresponding predicted SL structure. RNase probing of the WNV 3' minus-strand stem-loop RNA [WNV (-)3' SL RNA] confirmed the existence of a terminal secondary structure. RNA-protein binding studies were performed with BHK S100 cytoplasmic extracts and in vitro-synthesized WNV (-)3' SL RNA as the probe. Three RNA-protein complexes (complexes 1,2, and 3) were detected by a gel mobility shift assay, and the specificity of the RNA-protein interactions was confirmed by gel mobility shift and UV-induced cross-linking competition assays. Four BHK cell proteins with molecular masses of 108, 60, 50, and 42 kDa were detected by UV-induced cross-linking to the WNV (-)3' SL RNA. A preliminary mapping study indicated that all four proteins bound to the first 75 nucleotides of the WNV 3' minus-strand RNA, the region that contains the terminal SL. A flavivirus resistance phenotype was previously shown to be inherited in mice as a single, autosomal dominant allele. The efficiencies of infection of resistant cells and susceptible cells are similar, but resistant cells (C3H/RV) produce less genomic RNA than congenic, susceptible cells (C3H/He). Three RNA protein complexes and four UV-induced cross-linked cell proteins with mobilities identical to those detected in BHK cell extracts with the WNV (-)3' SL RNA were found in both C3H/RV and C3H/He cell extracts. However, the half-life of the C3H/RV complex 1 was three times longer than that of the C3H/He complex 1. It is possible that the increased binding activity of one of the resistant cell proteins for the flavivirus minus-strand RNA could result in a reduced synthesis of plus-strand RNA as observed with the flavivirus resistance phenotype. PMID- 8709256 TI - A seven-transmembrane domain receptor involved in fusion and entry of T-cell tropic human immunodeficiency virus type 1 strains. AB - Entry of human immunodeficiency virus type 1 (HIV-1) into cells requires binding to CD4 and fusion with a cellular membrane. Fusion does not occur in most nonhuman cells even when they express human CD4, indicating that one or more human accessory factors are required for virus infection. Recently, a seven transmembrane domain protein has been shown to serve as an accessory factor for T cell-tropic (T-tropic) HIV-1 isolates (Y. Feng, C. C. Broder, P. E. Kennedy, and E. A. Berger, Science 272:872-877, 1996). Here we show that expression of this glycoprotein, termed fusin, in murine, feline, simian, and quail cell lines, in conjunction with human CD4, rendered these cells fully permissive for HIV-1 envelope glycoprotein (Env)-mediated membrane fusion. Expression of CD4 or fusin alone did not permit fusion. In addition, introduction of fusin and CD4 into a human cell line, U87MG, that is resistant to HIV-1 induced syncytium formation and to infection by HIV-1 when expressing CD4 alone made this cell line permissive for Env-mediated cell-cell fusion. Fusion was observed only with T tropic Env proteins. Macrophage-tropic (M-tropic) Env proteins from the SF162, ADA, and Ba-L HIV-1 strains did not fuse with cells expressing fusin and CD4, suggesting that M-tropic viruses utilize an accessory molecule other than fusin. Finally, coexpression of fusin and CD4 made both a murine and feline cell line susceptible to virus infection by T-tropic, but not M-tropic, HIV-1 strains. PMID- 8709257 TI - Peripheral blood mononuclear cells from sheep infected with a variant of bovine leukemia virus synthesize envelope glycoproteins but fail to induce syncytia in culture. AB - Peripheral blood mononuclear cells (PBMCs) infected with the oncogenic retrovirus bovine leukemia virus (BLV) produce virus when cultured briefly. BLV can be transmitted in cocultures to adherent susceptible cells, which become infected, express viral proteins, and fuse into multinucleated syncytia several days later. PBMCs from 3 of 10 BLV-infected sheep displayed a lifelong deficiency in induction of syncytium formation among indicator cells in culture, although large numbers of PBMCs synthesized viral transcripts or capsid protein. Since the infected, syncytium-deficient PBMCs were > or = 97% B cells, the deficiency could not be attributed to altered host cell tropism. The syncytium-deficient phenotype was recapitulated in newly infected sheep, demonstrating that this property is regulated by the viral genotype. The alteration in the BLV genome delayed but did not prohibit the establishment of BLV infection in vivo. Envelope glycoproteins were synthesized in syncytium-deficient PBMCs, translocated to the cell surface, and incorporated into virions. However, monoclonal antibodies specific for the BLV surface glycoprotein did not stain fixed PBMCs of the syncytium-deficient phenotype. Moreover, an animal with syncytium-deficient PBMCs had lower titers of neutralizing antibodies throughout the first 5 years of infection than an animal with similar numbers of infected PBMCs of the syncytium-inducing phenotype. The syncytium-deficient variant productively infected indicator cells at greatly reduced efficiency, showing that the alteration affects an early step in viral entry or replication. These results suggest that the alteration maps in the env gene or in a gene whose product affects the maturation or conformation, and consequently the function, of the envelope protein complex. PMID- 8709258 TI - Activation of the human thymidine kinase (TK) promoter by simian virus 40 large T antigen requires both the T antigen pRb family-binding domain and TK promoter sequences resembling E2F-binding sites. AB - Infection of quiescent cells with the DNA tumor virus simian virus 40 induces expression of the cellular thymidine kinase (TK) gene a minimum of 10- to 20 fold, and this induction depends upon the viral protein large T antigen (T-Ag). To define both human TK promoter elements and T-Ag functional domains required for transcriptional induction, we have established a system in which stable Rat-1 transfectants harboring TK promoter-luciferase hybrid genes are infected with recombinant adenoviruses expressing either wild-type or mutant forms of T-Ag and luciferase expression is measured as an indicator of promoter activity. The results show that (i) a 135-bp TK promoter fragment is activated 10- to 15-fold by viral infection; (ii) this activation is the result of both T-Ag-dependent and -independent mechanisms; (iii) the T-Ag pRb family-binding domain, but not the p53-binding, helicase, or ATPase domain, is required for activation; and (iv) activation is severely diminished with a TK promoter fragment in which E2F-like binding sites have been removed. These data demonstrate a requirement for both an E2F-related factor and a pRb family member in activation of the TK promoter by T Ag. This contrasts with the promiscuous activation of many cellular and viral genes by T-Ag, which is independent of its ability to bind pRb. PMID- 8709259 TI - Human adenovirus-specific CD8+ T-cell responses are not inhibited by E3-19K in the presence of gamma interferon. AB - Adenovirus has considerable potential as a gene therapy vector, but recent animal data suggest that transduced cells are destroyed by adenovirus-specific cytotoxic T-lymphocyte (CTL) responses. Therefore, it will be important to develop strategies to evade adenovirus-specific CTL responses in humans. As a first step, an assay was developed to detect and characterize human CTLs directed against adenovirus. Adenovirus-specific CTL responses were demonstrated to be present in four of five healthy adults by in vitro stimulation of peripheral blood mononuclear cells with autologous fibroblasts infected with the adenovirus type 2 (Ad2) E3 deletion mutant Ad2+ND1. Killing by adenovirus-specific CTLs was major histocompatibility complex class I restricted and was documented to be mediated by CD8+ T cells. Wild-type-Ad2-infected cells were poor CTL targets compared with cells infected with the E3 deletion mutant because of the expression of E3-19K, an early viral glycoprotein which prevents transport of major histocompatibility complex class I antigens out of the endoplasmic reticulum to the cell surface. However, preincubation of targets with gamma interferon resulted in enhanced killing of wild-type-Ad2-infected cells, to levels comparable to those obtained with Ad2+ ND1-infected cells. Radioimmunoprecipitation analysis revealed that gamma interferon not only increased the synthesis of class I antigens but also allowed excess molecules to escape from the endoplasmic reticulum. It is concluded that E3-19K expression in adenovirus-infected cells inhibits human CTL recognition in vitro but that gamma interferon may help overcome the E3-19K effect during acute infection in vivo. PMID- 8709261 TI - Inhibition of herpes simplex virus type 1 immediate-early gene expression by alpha interferon is not VP16 specific. AB - Pretreatment of tissue culture cells with alpha interferon (IFN-alpha) inhibits the transcription of herpes simplex virus type 1 (HSV-1) immediate-early (IE) genes, an effect which has been attributed to reduced transactivation of IE promoters by the virion protein VP16. Our previous demonstration that IFN-alpha inhibited the replication of the HSV-1 mutant in1814, which has a mutated VP16 unable to activate IE transcription, appeared to be incompatible with IFN-alpha having an effect on VP16 action (D. R. S. Jamieson, L. H. Robinson, J. I. Daksis, M. J. Nicholl, and C. M. Preston, J. Gen. Virol. 76:1417-1431, 1995). To investigate this observation further, cells were infected with a derivative of in1814 containing the lacZ gene controlled by the human cytomegalovirus IE promoter. The accumulation of HSV-1 IE RNA species was inhibited by IFN-alpha in these cells to the same extent as in cells infected with a virus rescued at the VP16 locus, and production of lacZ-specific RNA was also reduced, demonstrating that IFN-alpha can inhibit expression from a heterologous promoter that is not responsive to VP16. To provide a means of investigating the activity of VP16 on IE promoters not located in the HSV-1 genome, cell lines containing the neomycin phosphotransferase gene controlled by the HSV-1 IE ICPO promoter were constructed. Activation of the IE promoter by VP16 was not inhibited when the ICPO promoter was resident in the cell, demonstrating that VP16 function was unaffected by pretreatment of cells with IFN-alpha. The results suggest that IFN alpha prevents the onset of IE transcription from the HSV-1 genome through a general mechanism rather than by having an effect specific to HSV-1 IE promoters. PMID- 8709260 TI - Adenovirus early region 4 34-kilodalton protein directs the nuclear localization of the early region 1B 55-kilodalton protein in primate cells. AB - The localization of the adenovirus type 5 34-kDa E4 and 55-kDa E1B proteins was determined in the absence of other adenovirus proteins. When expressed by transfection in human, monkey, hamster, rat, and mouse cell lines, the E1B protein was predominantly cytoplasmic and typically was excluded from the nucleus. When expressed by transfection, the E4 protein accumulated in the nucleus. Strikingly, when coexpressed by transfection in human, monkey, or baby hamster kidney cells, the E1B protein colocalized in the nucleus with the E4 protein. A complex of the E4 and E1B proteins was identified by coimmunoprecipitation in transfected HeLa cells. By contrast to the interaction observed in primate and baby hamster kidney cells, the E4 protein failed to direct the E1B protein to the nucleus in rat and mouse cell lines as well as CHO and V79 hamster cell lines. This failure of the E4 protein to direct the nuclear localization of the E1B protein in REF-52 rat cells was overcome by fusion with HeLa cells. Within 4 h of heterokaryon formation and with protein synthesis inhibited, a portion of the E4 protein present in the REF-52 nuclei migrated to the HeLa nuclei. Simultaneously, the previously cytoplasmic E1B protein colocalized with the E4 protein in both human and rat cell nuclei. These results suggest that a primate cell-specific factor mediates the functional interaction of the E1B and E4 proteins of adenovirus. PMID- 8709262 TI - The UL97 gene product of human cytomegalovirus is an early-late protein with a nuclear localization but is not a nucleoside kinase. AB - The temporal expression of the UL97 gene product during human cytomegalovirus (HCMV) infection of human foreskin fibroblasts (HFF) and subcellular localization of this protein were analyzed by using a polyclonal antiserum raised against a truncated UL97 protein of 47 kDa. The UL97 protein was detectable 16 h after infection by Western blot (immunoblot) analysis. Since only reduced UL97 expression occurred in the presence of two inhibitors of DNA replication, phosphonoacetic acid and ganciclovir, we conclude that UL97 is an early-late gene, requiring DNA replication for maximum expression. By indirect immunofluorescence, the protein could be visualized in the nuclei of virus infected HFF 22 h after infection. Nuclear localization of the UL97 protein was also detected in thymidine kinase-deficient 143B cells infected with a recombinant vaccinia virus containing the entire UL97 open reading frame (ORF), as well as in HFF transiently expressing the entire UL97 ORF under the control of HCMV major immediate-early promoter. However, transiently expressed 5'-terminal deletion mutants of the UL97 ORF in addition showed a cytoplasmic localization of the UL97 protein, confirming the presence of a nuclear localization site in the N terminal region of the protein. Our high-pressure liquid chromatography analyses confirmed the ganciclovir phosphorylation by the UL97 protein, but no specific phosphorylation of natural nucleosides was observed, indicating that the UL97 protein is not a nucleoside kinase. During plaque purification of recombinant UL97-deficient HCMV, this virus was growth defective; hence, we presume that UL97 may be essential for the viral life cycle. PMID- 8709263 TI - Localization of human T-cell leukemia virus type 1 tax to subnuclear compartments that overlap with interchromatin speckles. AB - Tax, the virally encoded activator of the human T-cell leukemia virus type 1 long terminal repeats, regulates the expression of many cellular genes. This protein has been implicated in transformation events leading to the development of adult T-cell leukemia. Because subcellular localization contributes importantly to protein function, we determined the compartment(s) within the cell in which Tax is found. Using confocal microscopy, we found that Tax localizes to subnuclear domains which overlap with structures previously identified as interchromatin granules or spliceosomal speckles. These Tax speckled structures are coincident with a subset of nuclear transcriptional hot spots. Disruption of the Tax speckled structures by heat shock revealed the existence of different populations of Tax. One population of Tax is tightly associated with nuclear speckles. A second population exists outside of the speckles and is transcriptionally active for some promoters. PMID- 8709264 TI - Prolonged gene expression and cell survival after infection by a herpes simplex virus mutant defective in the immediate-early genes encoding ICP4, ICP27, and ICP22. AB - Very early in infection, herpes simplex virus (HSV) expresses four immediate early (IE) regulatory proteins, ICP4, ICP0, ICP22, and ICP27. The systematic inactivation of sets of the IE proteins in cis, and the subsequent phenotypic analysis of the resulting mutants, should provide insights into how these proteins function in the HSV life cycle and also into the specific macromolecular events that are altered or perturbed in cells infected with virus strains blocked very early in infection. This approach may also provide a rational basis to assess the efficacy and safety of HSV mutants for use in gene transfer experiments. In this study, we generated and examined the phenotype of an HSV mutant simultaneously mutated in the ICP4, ICP27, and ICP22 genes of HSV. Unlike mutants deficient in ICP4 (d120), ICP4 and ICP27 (d92), and ICP4 and ICP22 (d96), mutants defective in ICP4, ICP27, and ICP22 (d95) were visually much less toxic to Vero and human embryonic lung cells. Cells infected with d95 at a multiplicity of infection of 10 PFU per cell retained a relatively normal morphology and expressed genes from the viral and cellular genomes for at least 3 days postinfection. The other mutant backgrounds were too toxic to allow examination of gene expression past 1 day postinfection. However, when cell survival was measured by the capacity of the infected cells to form colonies, d95 inhibited colony formation similarly to d92. This apparent paradox was reconciled by the observation that host cell DNA synthesis was inhibited in cells infected with d120, d92, d96, and d95. In addition, all of the mutants exhibited pronounced and distinctive alterations in nuclear morphology, as determined by electron microscopy. The appearance of d95-infected cells deviated from that of uninfected cells in that large circular structures formed in the nucleus. d95-infected cells abundantly expressed ICP0, which accumulated in fine punctate structures in the nucleus at early times postinfection and coalesced or grew to the large circular objects that were revealed by electron microscopy. Therefore, while the abundant accumulation of ICPO in the absence of ICP4, ICP22, and ICP27 may allow for prolonged gene expression, cell survival is impaired, in part, as a result of the inhibition of cellular DNA synthesis. PMID- 8709265 TI - Transient subversion of CD40 ligand function diminishes immune responses to adenovirus vectors in mouse liver and lung tissues. AB - First-generation adenovirus vectors will have limited application in gene therapy for chronic diseases because of destructive host immune responses. Important immune effectors include CD8+ T cells, which mediate target cell destruction and ablate transgene expression, and B cells, which produce neutralizing antibodies that block effective readministration of vector. Previous studies indicated that activation of CD4+ T cells by virus capsid proteins is necessary for full realization of effector function of CD8+ T cells and B cells. In this paper, we present a strategy for preventing CD4+ T-cell activation by an adenovirus vector delivered to mouse liver and lung tissues which is based on interfering with T cell priming via CD40 ligand-CD40 interactions. Adenovirus transgene expression was stabilized in mice genetically deficient in CD40 ligand (CD40L), and neutralizing antibody to adenovirus did not develop, allowing efficient readministration of vector. A transient blockade of T-cell activation with an antibody to CD40L infused into the animal at the time of adenovirus vector mediated gene transfer led to stabilization of transgene expression and diminished production of neutralizing antibody, allowing readministration of vector. In vitro T-cell assays suggested that a block in the primary activation of CD4+ T cells was responsible for the lack of B-cell- and cytotoxic-T-cell dependent responses. This suggests a strategy for improving the potential of adenovirus vectors based on administration of an antibody to CD40L at the time of vector administration. PMID- 8709266 TI - The 126- and 183-kilodalton proteins of tobacco mosaic virus, and not their common nucleotide sequence, control mosaic symptom formation in tobacco. AB - Nucleotide substitutions at two positions within the open reading frame encoding the 126-kDa protein in the attenuated masked (M) strain of tobacco mosaic tobamovirus (TMV) to those found in the virulent U1-TMV genome led to the induction of near U1-TMV-like symptoms on leaves of Nicotiana tabacum L. cv. Xanthi nn by progeny virus (M. H. Shintaku, S. A. Carter, Y. Bao, and R. S. Nelson, Virology 221:218-225, 1996). In this study, further site-directed mutations were made at these positions within the M strain cDNA to determine whether the protein or nucleotide sequence directly controlled the symptom phenotype. The protein and not the nucleotide sequence directly controlled the symptom phenotype when amino acid 360 within the 126-kDa protein sequence was altered and likely controlled the symptom phenotype when amino acid 601 was altered. The effects of the substitutions at amino acid position 360 on viral protein accumulation were studied by pulse-labeling proteins in infected protoplasts. Accumulation of the 126- and 183-kDa proteins was less for an attenuated mutant than for two virulent mutants, but the viral movement protein and coat protein accumulated to levels reported to be sufficient for normal systemic symptom development. The size of necrotic local lesions on N. tabacum L. cv. Xanthi NN was negatively correlated with symptom development and accumulation of the 126-kDa protein for these mutants. With reference to this last finding, an explanation of the cause of the differing symptoms induced by these viruses is presented. PMID- 8709267 TI - Identification of domains in the simian immunodeficiency virus matrix protein essential for assembly and envelope glycoprotein incorporation. AB - The matrix domain (MA) of the simian immunodeficiency virus (SIV) is encoded by the amino-terminal region of the Gag polyprotein precursor and is the component of the viral capsid that lines the inner surface of the virus envelope. To define domains of the SIV MA protein that are involved in virus morphogenesis, deletion and substitution mutations were introduced in this protein in the context of a gag-protease construct and expressed in the vaccinia virus vector system. The MA mutants were characterized with respect to synthesis and processing of the Gag precursor, assembly and release of virus-like particles, and incorporation of the envelope (Env) glycoprotein into particles. We have identified two regions of the SIV MA which are critical for particle formation. Both domains are located in a central hydrophobic alpha-helix of the SIV MA, according to data on the structure of this protein. In addition, we have characterized a domain whose mutation impairs the incorporation of SIV Env glycoproteins with long transmembrane cytoplasmic tails into particles. Interestingly, these mutant particles retained the ability to associate with SIV Env proteins with short cytoplasmic tails. PMID- 8709268 TI - The PB1 subunit alone can catalyze cRNA synthesis, and the PA subunit in addition to the PB1 subunit is required for viral RNA synthesis in replication of the influenza virus genome. AB - We indicated that the PB1 and PA subunits of RNA polymerase and nucleoprotein (NP) can support replication of the influenza virus genome as well as transcription to yield uncapped poly(A)(+)-RNA (Y. Nakagawa, N. Kimura, T. Toyoda, K. Mizumoto, A. Ishihama, K. Oda, and S. Nakada, J. Virol. 69:728-733, 1995). To analyze the functions of the PB1 and PA subunits in replication and transcription, YP1N clones in which the PB1 and NP genes can be expressed in response to dexamethasone were established. cRNA was transcribed from model viral RNA (vRNA), but vRNA synthesis from model cRNA was not detected in YP1N clones. Furthermore, poly(A)(+)-RNA directed from model vRNA was synthesized in YP1N clones. These results indicated that PB1 and NP can support the syntheses of cRNA and poly(A)(+)-RNA and that the PA subunit, in addition to that of PB1 and to NP, is required for vRNA synthesis. In summary, the PB1 subunit is involved in the catalytic activities of nucleotide elongation, and the PA subunit may act as an allosteric modulator and cause a conformational change from a cRNA-to a vRNA synthesizing form of the PB1 subunit. PMID- 8709269 TI - Persistent poliovirus infection of human fetal brain cells. AB - It has been suggested that poliovirus (PV), the causative agent of poliomyelitis, could persist in surviving patients. We have previously shown that PV can persistently infect some human cell lines in vitro, particularly neuroblastoma cell lines. We report here an ex vivo model in which PV can persistently infect primary cultures of human fetal brain cells. Two mutations involving capsid residues 142 of VP2 and 95 of VP1 were repeatedly selected during the persistent infections. These residues are located in capsid regions known to be involved in interactions between PV and its receptor. During the first week after infection, viral antigens were found in cells of both the neuronal and glial lineages. In contrast, 2 weeks after infection, viral antigens were detected almost exclusively in cells of the neuronal lineage. They were detected predominantly in cells expressing a marker of early commitment to the neuronal lineage, MAP-5, particularly in neuroblasts. Viral antigens were also found in immature progenitors expressing a neuroepithelium marker, nestin, and in cells expressing a marker of postmitotic neurons, MAP-2. The presence of viral antigens in postmitotic neurons suggests that PV can persist in neurons of patients who have survived poliomyelitis. PMID- 8709270 TI - Exogenous thymidine is preferentially incorporated into human cytomegalovirus DNA in infected human fibroblasts. AB - The effect of human cytomegalovirus infection on cellular DNA synthesis in human fibroblasts was measured by fluorometry and by incorporation of radiolabeled thymidine. The results show that although HCMV infection stimulates cellular DNA synthesis in both quiescent and serum-stimulated cells, radiolabeled thymidine is almost exclusively incorporated into viral DNA. PMID- 8709271 TI - Possible origin of murine AIDS (MAIDS) virus: conversion of an endogenous retroviral p12gag sequence to a MAIDS-inducing sequence by frameshift mutations. AB - The murine AIDS (MAIDS) virus has a unique sequence in its p12gag region, which is responsible for MAIDS development. A transcript hybridizing with this sequence is expressed in normal C57BL/6 mice. The transcript, designated Edv, has been previously cloned and sequenced (Y. Kubo, Y. Nakagawa, K. Kakimi, H. Matsui, K. Higo, L. Wang, H. Kobayashi, T. Hirama, and A. Ishimoto, J. Gen. Virol. 75:881 888, 1994). Compared with the nucleotide sequence of the helper LP-BM5 ecotropic virus, the pathogenic replication-defective MAIDS virus has a 16-bp deletion and a 1-bp insertion in the 5' and 3' regions of the p12gag sequence, respectively, and the Edv transcript contains only a 3-bp deletion. Therefore, the amino acid sequence of the defective MAIDS virus p12gag region is not homologous to that of the helper virus and the Edv transcript because of the frameshift. To determine whether the amino acid sequence resulting from the frameshift is critical for MAIDS development, we constructed chimeric viruses that contained the p12gag regions of the helper virus and the Edv transcript, respectively, with and without the same frame as the defective MAIDS virus by the artificial frameshift mutations. The mutant viruses with the frameshift mutations induced MAIDS in inoculated mice, but the viruses without the mutations did not. These results suggested that the MAIDS virus was generated by frameshift mutations in the p12gag region of Edv or a related sequence. PMID- 8709272 TI - Virulence as a positive trait in viral persistence. AB - A population replacement experiment has been devised to test the ability of a challenge virus to replace the resident virus in a persistently infected cell culture. BHK-21 cells persistently infected with foot-and-mouth disease virus of serotype C (clone C-S8c1) were challenged with a large excess of either the parental foot-and-mouth disease virus C-S8c1, genetically marked variants differing in their degree of virulence, or a mutant rescued after prolonged persistence in BHK-21 cells. After challenge, the composition of the resident virus population in the carrier culture was analyzed by reverse transcription-PCR amplification and nucleotide sequencing. The dominance of the initial persisting virus was seen in all cases, except when virulent viruses were used in the challenge. The experiments document that, paradoxically, virulence can be a positive factor in the reestablishment of a virus population in a persistently infected cell culture. A model based on the selection of virus-resistant cell variants during persistence is proposed to interpret these observations. Implications about the persistence of viruses in their host cells and organisms are discussed. PMID- 8709273 TI - Large-population passages of vesicular stomatitis virus in interferon-treated cells select variants of only limited resistance. AB - Vesicular stomatitis virus (VSV) populations were repeatedly passaged in L-929 cells treated with alpha interferon (IFN-alpha) at levels of 25 U/ml. This IFN alpha concentration induced a 99.9% inhibition of viral yield in standard infections. Analysis of viral fitness (overall replicative ability measured in direct competition with a reference wild-type VSV) after 21 passages in IFN treated cells showed only a limited increase or no increase in fitness, compared with the greater increase upon parallel passage in cells not treated with IFN alpha. However, this limited increase in fitness was more pronounced when competition assays were carried out with IFN-alpha-treated cells, suggesting the selection of VSV populations with a low level of resistance to IFN-alpha. Thus, despite the extensively documented capacity of VSV to adapt to changing environments, the antiviral state induced by IFN-alpha imposes adaptive constraints on VSV which are not readily overcome. PMID- 8709275 TI - Revertant analysis of J-K mutations in the encephalomyocarditis virus internal ribosomal entry site detects an altered leader protein. AB - The internal ribosomal entry site (IRES) of picornaviruses consists of various sequence and structural elements that collectively impart translational function to the genome. By engineering substitution and deletion mutations into the J-K elements of the encephalomyocarditis virus IRES, translationally defective viruses with small-plaque phenotypes were generated. From these, 60 larger-plaque revertant viruses were isolated and characterized, and their sequences were compared with a structural model of the IRES. The data provide confirming evidence for the existence of helix J3 within stem J but suggest that helix J1 is 3 bp longer than previously estimated. They also suggest that previously modeled stems L and M should be replaced by an alternative structure. One reversion mutation was mapped to the leader protein coding region. This change of leader amino acid 20 from Pro to Ser increased the viral plaque size dramatically but did not alter the cell-free translational activity of the mutated, parental IRES. PMID- 8709274 TI - Oral immunization with a replication-deficient recombinant vaccinia virus protects mice against influenza. AB - Mice immunized with two intragastrically administered doses of a replication deficient recombinant vaccinia virus containing the hemagglutinin and nucleoprotein genes from H1N1 influenza virus developed serum anti-H1 immunoglobulin G (IgG) antibody that completely protected the lungs from challenge with H1N1. Almost all of the mice given two intragastric doses also developed mucosal anti-H1 IgA antibody, and those with high anti-H1 IgA titers had completely protected noses. Intramuscular injection of the vaccine protected the lungs but not the noses from challenge. We also found that the vaccine enhanced recovery from infection caused by a shifted (H3N2) influenza virus, probably through the induction of nucleoprotein-specific cytotoxic T-lymphocyte activity. A replication-deficient, orally administered, enteric-coated, vaccinia virus-vectored vaccine might safely protect humans against influenza. PMID- 8709276 TI - Differential glycosylation, virion incorporation, and sensitivity to neutralizing antibodies of human immunodeficiency virus type 1 envelope produced from infected primary T-lymphocyte and macrophage cultures. AB - Two primary cell targets for human immunodeficiency virus type 1 (HIV-1) infection in vivo are CD4+ T lymphocytes and monocyte-derived macrophages (MDM). HIV-1 encodes envelope glycoproteins which mediate virus entry into these cells. We have utilized infected and radiolabelled primary peripheral blood mononuclear cell (PBMC) and MDM cultures to examine the biochemical and antigenic properties of the HIV-1 envelope produced in these two cell types. The gp120 produced in MDM migrates as a broad, diffuse band in sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels compared with that of the more homogeneous gp120 released from PBMCs. Glycosidase analyses indicated that the diffuse appearance of the MDM gp120 is due to the presence of asparagine-linked carbohydrates containing lactosaminoglycans, a modification not observed with the gp120 produced in PBMCs. Neutralization experiments, using isogeneic PBMC and MDM-derived macrophage tropic HIV-1 isolates, indicate that 8- to 10-fold more neutralizing antibody, directed against the viral envelope, is required to block virus produced from MDM. These results demonstrate that HIV-1 released from infected PBMC and MDM cultures differs in its biochemical and antigenic properties. PMID- 8709277 TI - Human immunodeficiency virus type 1 membrane fusion mediated by a laboratory adapted strain and a primary isolate analyzed by resonance energy transfer. AB - Previous studies of human immunodeficiency virus type 1 (HIV-1) envelope glycoprotein-mediated membrane fusion have focused on laboratory-adapted T lymphotropic strains of the virus. The goal of this study was to characterize membrane fusion mediated by a primary HIV-1 isolate in comparison with a laboratory-adapted strain. To this end, a new fusion assay was developed on the basis of the principle of resonance energy transfer, using HeLa cells stably transfected with gp120/gp41 from the T-lymphotropic isolate HIV-1LA1 or the macrophage-tropic primary isolate HIV-1JR-FL. These cells fused with CD4+ target cell lines with a tropism mirroring that of infection by the two viruses. Of particular note, HeLa cells expressing HIV-1JR-FL gp120/gp41 fused only with PM1 cells, a clonal derivative of HUT 78, and not with other T-cell or macrophage cell lines. These results demonstrate that the envelope glycoproteins of these strains play a major role in mediating viral tropism. Despite significant differences exhibited by HIV-1JR-FL and HIV-1LAI in terms of tropism and sensitivity to neutralization by CD4-based proteins, the present study found that membrane fusion mediated by the envelope glycoproteins of these viruses had remarkably similar properties. In particular, the degree and kinetics of membrane fusion were similar, fusion occurred at neutral pH and was dependent on the presence of divalent cations. Inhibition of HIV-1JR-FL envelope glycoprotein mediated membrane fusion by soluble CD4 and CD4-IgG2 occurred at concentrations similar to those required to neutralize this virus. Interestingly, higher concentrations of these agents were required to inhibit HIV-1LAI envelope glycoprotein-mediated membrane fusion, in contrast to the greater sensitivity of HIV-1LAI virions to neutralization by soluble CD4 and CD4-IgG2. This finding suggests that the mechanisms of fusion inhibition and neutralization of HIV-1 are distinct. PMID- 8709278 TI - The human T-cell leukemia virus type 1 posttranscriptional trans-activator Rex contains a nuclear export signal. AB - The Rex protein of human T-cell leukemia virus type 1 is required for the nuclear export of unspliced viral mRNA and, therefore, for virus replication. In this manuscript, we demonstrate that Rex shuttles between the nucleus and the cytoplasm and that its activation domain constitutes a nuclear export signal that specifies efficient transport to the cytoplasm. These findings are consistent with a model for Rex-mediated trans-activation in which Rex-viral mRNA complexes are targeted for nuclear export by the direct action of the activation domain. PMID- 8709279 TI - DNA double-strand break repair functions defend against parvovirus infection. AB - We measured parvovirus replication and sensitivity to X-ray damage in nine CHO cell lines representing a variety of DNA repair deficiencies. We found that parvovirus replication efficiency increases with radiosensitivity. Parvovirus replication is disrupted at an early stage of infection in DNA repair-proficient cells, before conversion of the single-stranded viral DNA genome into the double stranded replicative form. Thus, status of the DNA repair machinery inversely correlates with parvovirus replication and is proportional to the host's ability to repair X-ray-induced damage. PMID- 8709280 TI - Genetics of intracisternal-A-particle-related envelope-encoding proviral elements in mice. AB - Intracisternal-A-particle-related envelope-encoding (IAPE) proviral elements in the mouse genome encode and express an envelope-like protein that may allow transmission of IAPEs as infectious agents. To test IAPE mobility and potential transmission in mice, we have analyzed the distribution of IAPE elements in the genomes of Mus spretus and Mus musculus inbred strains and wild-caught animals. Potential full-length (IAPE-A) proviral elements are present as repetitive copies in DNA from male but not female animals of M. musculus inbred strains and Mus musculus castaneus. Analysis of IAPE-cellular junction fragments indicates that fixation of most IAPEs in the germ line occurred in M. musculus and M. spretus after speciation but before M. musculus inbred strains were derived. PMID- 8709281 TI - Relationship between viral DNA synthesis and virion envelopment in hepatitis B viruses. AB - While the intracellular pool of encapsidated hepatitis B viral DNA contains genomes in all stages of DNA replication, serum-derived virions contain predominantly mature, partially duplex, circular DNA genomes. To account for this finding, Summers and Mason proposed in 1982 that virion envelopment is somehow linked to the state of genomic maturation (J. Summers and W.S. Mason, Cell 29:403 415, 1982). Core gene mutations with phenotypes consistent with this concept have previously been identified in the duck hepatitis B virus (DHBV). Here we show that DHBV polymerase mutants with altered DNA synthesis also display defects in envelopment, and we provide quantitative estimates of the magnitude of the preference for the envelopment of mature DNA. In cells transfected with wild-type DHBV DNA, immature minus-strand DNA represents 18% of the intracellular pool but only 4% of extracellular virion DNA. A point mutation in the C-terminal domain of the polymerase strongly and selectively impairs plus-strand synthesis; in this mutant, the ratio of immature to mature DNA in the intracellular pool rises to 6:1 but is reduced to 1.5:1 in released virions. A missense mutation in the polymerase active site inactivates all viral DNA synthesis but still allows efficient RNA encapsidation; in this mutant, no detectable viral nucleic acid is enveloped and released. Thus, viral DNA synthesis is absolutely required for envelopment and export, and a strong further bias exists in favor of the export of genomes that have completed minus-strand synthesis and at least initiated plus strand synthesis. These results imply that events within the interior of the nucleocapsid can powerfully influence its interactions with external viral envelope glycoproteins. PMID- 8709282 TI - Sequence analysis of the human DNA flanking sites of human immunodeficiency virus type 1 integration. AB - Human immunodeficiency virus type 1 (HIV-1) tagged with the Escherichia coli supF gene has been used to clone integrated HIV-1 proviruses. Sequence analysis of the 600 to 800 bp of human DNA adjacent to 29 clones revealed a propensity for HIV-1 to integrate near the Alu class of human repetitive elements. PMID- 8709283 TI - Interaction of the adenovirus L1 52/55-kilodalton protein with the IVa2 gene product during infection. AB - The adenovirus L1 52/55-kDa protein is expressed both in the early and late stages of infection, raising the possibility that it has multiple roles in the viral life cycle. To obtain possible insights into these roles, the yeast two hybrid system was used to examine the interactions of the 52/55-kDa protein with viral and cellular factors. cDNA expression libraries from human 293 cells at both early and late stages of adenovirus type 5 infection were constructed and screened, with the 52/55-kDa protein being used as bait. Characterization of positive clones revealed that the adenovirus IVa2 gene product interacted specifically with the 52/55-kDa protein. In addition, the IVa2 protein was shown to interact with a bacterial glutathione S-transferase-52/55-kDa fusion protein in vitro, further supporting the finding with the yeast two-hybrid system. Finally, coimmunoprecipitation studies confirmed that the 52/55-kDa protein and IVa2 polypeptide interact specifically during the course of adenovirus infection. A potential role for the IVa2-52/55-kDa protein interaction in the regulation of transcription from the major late promoter and in viral assembly is discussed. PMID- 8709284 TI - Proteoglycans secreted by packaging cell lines inhibit retrovirus infection. AB - Using a model recombinant retrovirus encoding the Escherichia coli lacZ gene, we have found that medium conditioned with NIH 3T3 cells and packaging cell lines derived from NIH 3T3 cells inhibits infection. Most of the inhibitory activity was greater than 100 kDa and was sensitive to chondroitinase ABC digestion, which is consistent with the inhibitor being a chondroitin sulfate proteoglycan. Proteoglycans secreted by NIH 3T3 cells and purified by anion-exchange chromatography inhibited amphotropic retrovirus infection. Pretreatment of amphotropic retrovirus stocks with chondroitinase ABC boosted the level of transduction efficiency by more than twofold. The implications of these findings with respect to retrovirus-cell interactions and the production of high-titer retroviral stocks are discussed. PMID- 8709285 TI - The Moloney murine sarcoma virus ts110 5' splice site signal contributes to the regulation of splicing efficiency and thermosensitivity. AB - The 5' splice site signal (5'ss) in Moloney murine sarcoma virus ts110 (MuSVts110) RNA was found to participate in the regulation of its splicing phenotype. This 5'ss (CAG/GUAGGA) departs from the mammalian consensus (CAG/GURAGU) at positions +4 and +6, both of which base pair with U1 and U6 small nuclear RNAs during splicing. A doubling in splicing efficiency and near elimination of the splicing thermosensitivity characteristic of MuSVts110 were observed in 5'ss mutants containing a U at position +6 (termed 5' A6U), even in those in which U1-5'ss complementarity had been reduced. At the permissive temperature (28 degrees C), the 5' A6U mutation increased the efficiency of the second splicing reaction, while at the nonpermissive temperature (39 degrees C), both splicing reactions were positively affected. PMID- 8709286 TI - Protection against apoptosis by the vaccinia virus SPI-2 (B13R) gene product. AB - Vaccinia virus contains a gene, termed SPI-2 or B13R, that is closely related in its sequence to a potent inhibitor of apoptosis from cowpox virus (crmA). Infection by vaccinia virus protects HeLa cells against apoptosis that is induced by an immunoglobulin M antibody against the fas receptor or by tumor necrosis factor alpha. This effect is profoundly reduced when the SPI-2 gene is deleted. The SPI-2 gene, when transiently expressed in these cells, can also protect against apoptosis mediated by these agents. Given the similarity to crmA, it seems likely that SPI-2 functions in an analogous fashion, inhibiting the activity of ICE protease family members and blocking the onset of apoptosis. PMID- 8709287 TI - Correlation between poliovirus type 1 Mahoney replication in blood cells and neurovirulence. AB - Poliovirus (PV) is not often described as a monocyte- or macrophage-tropic virus; however, previous work indicated that neurovirulent PV type 1 Mahoney [PV(1)Mahoney] can productively infect primary human monocytes. To determine whether this replication has a functional role in pathogenesis, primary human mononuclear blood cells were infected with pairs of attenuated and neurovirulent strains of PV. Two neurovirulent strains of PV, PV(1)Mahoney and PV(2)MEF-1, replicated faster and to higher titers than attenuated counterparts PV(1)Sabin and PV(2)W-2, respectively, in primary human monocytes, suggesting that this replication may contribute to pathogenesis. PV(3)Leon grew weakly, while PV(3)Sabin, PV(2)Sabin, and PV(2) P712 did not replicate in these cells, perhaps because of their slow replication cycle. In U937 cells, a monocytelike cell line, PV(1)Mahoney replicated but PV(1)Sabin did not, while both grew well in HeLa cells. When molecular recombinants of PV(1)Mahoney and PV(1)Sabin were assessed, a correlation between neurovirulence and the ability to replicate in primary human mononuclear blood cells was found. Surprisingly, infectious centers assays with primary human mononuclear blood cells and U937 cells indicated that despite the lower overall viral yield, more cells are initially infected with the attenuated viruses. These results indicate that there are virulence-specific differences in the ability of PV(1)Mahoney to replicate in monocytes and suggest that there may be factors in monocytes that virulent strains of PV require. PMID- 8709288 TI - The association of Nef with a cellular serine/threonine kinase and its enhancement of infectivity are viral isolate dependent. AB - The nef genes of human immunodeficiency virus type 1 (HIV-1) and simian immunodeficiency virus (SIV) encode a 27- to 34-kDa myristoylated protein which induces downregulation of CD4 surface levels and enhances virus infectivity. In adult macaques, Nef has been implicated in pathogenesis and disease progression. Both HIV-1 SF2 Nef and SIVmac239 Nef have been shown to associate with a cellular serine/threonine kinase. We tested five functional Nef isolates to examine whether this kinase association is a property conserved among different isolates. HIV-1 SF2 and 248 and SIVmac239 Nef proteins were found associated with the kinase. HIV-1 NL4-3 and 233 Nef proteins were found weakly associated or not associated with the kinase. All five Nef isolates efficiently downregulated CD4 cell surface expression, suggesting that the association with this cellular kinase is not required for Nef to downregulate CD4. Comparison of the SF2 and NL4 3 isolates shows a differential ability of Nef to enhance infectivity that suggests a possible correlation between kinase association and enhancement of infectivity. PMID- 8709289 TI - A gene transfer vector-cell line system for complete functional complementation of adenovirus early regions E1 and E4. AB - The improvements to adenovirus necessary for an optimal gene transfer vector include the removal of virus gene expression in transduced cells, increased transgene capacity, complete replication incompetence, and elimination of replication-competent virus that can be produced during the growth of first generation adenovirus vectors. To achieve these aims, we have developed a vector cell line system for complete functional complementation of both adenovirus early region 1 (E1) and E4. A library of cell lines that efficiently complement both E1 and E4 was constructed by transforming 293 cells with an inducible E4-ORF6 expression cassette. These 293-ORF6 cell lines were used to construct and propagate viruses with E1 and E4 deleted. While the construction and propagation of AdRSV beta gal.11 (an E1-/E4- vector engineered to contain a deletion of the entire E4 coding region) were possible in 293-ORF6 cells, the yield of purified virus was depressed approximately 30-fold compared with that of E1- vectors. The debilitation in AdRSV beta gal.11 vector growth was found to correlate with reduced fiber protein and mRNA accumulation. AdCFTR.11A, a modified E1-/E4- vector with a spacer sequence placed between late region 5 and the right inverted terminal repeat, efficiently expressed fiber and grew with the same kinetic profile and virus yield as did E1- vectors. Moreover, purified AdCFTR.11A yields were equivalent to E1- vector levels. Since no overlapping sequences exist in the E4 regions of E1-/E4- vectors and 293-ORF6 cell lines, replication-competent virus cannot be generated by homologous recombination. In addition, these second generation E1-/E4- vectors have increased transgene capacity and have been rendered virus replication incompetent outside of the new complementing cell lines. PMID- 8709290 TI - Resistance of chimpanzee T cells to human immunodeficiency virus type 1 Tat enhanced oxidative stress and apoptosis. AB - CD4+ T-cell depletion in AIDS patients involves induction of apoptosis in human immunodeficiency virus (HIV)-infected and noninfected T cells. The HIV type 1 (HIV-1)-transactivating protein Tat enhances apoptosis and activation-induced cell death (AICD) of human T cells. This effect is mediated by the CD95 (APO 1/Fas) receptor-CD95 ligand (CD95L) system and may be linked to the induction of oxidative stress by Tat. Here we show that HIV-1 Tat-induced oxidative stress is necessary for sensitized AICD in T cells caused by CD95L expression. Tat-enhanced apoptosis and CD95L expression in T cells are inhibited by neutralizing anti-Tat antibodies, antioxidants, and the Tat inhibitor Ro24-7429. Chimpanzees infected with HIV-1 show viral replication resembling early infection in humans but do not show T-cell depletion or progression towards AIDS. The cause for this discrepancy is unknown. Here we show that unlike Tat-treated T cells in humans, Tat-treated chimpanzee T cells do not show downregulation of manganese superoxide dismutase or signs of oxidative stress. Chimpanzee T cells are also resistant to Tat enhanced apoptosis, AICD, and CD95L upregulation. PMID- 8709291 TI - Transmembrane domain of influenza virus neuraminidase, a type II protein, possesses an apical sorting signal in polarized MDCK cells. AB - The influenza virus neuraminidase (NA), a type II transmembrane protein, is directly transported to the apical plasma membrane in polarized MDCK cells. By using deletion mutants and chimeric constructs of influenza virus NA with the human transferrin receptor, a type II basolateral transmembrane protein, we investigated the location of the apical sorting signal of influenza virus NA. When these mutant and chimeric proteins were expressed in stably transfected polarized MDCK cells, the transmembrane domain of NA, and not the cytoplasmic tail, provided a determinant for apical targeting in polarized MDCK cells and this transmembrane signal was sufficient for sorting and transport of the ectodomain of a reporter protein (transferrin receptor) directly to the apical plasma membrane of polarized MDCK cells. In addition, by using differential detergent extraction, we demonstrated that influenza virus NA and the chimeras which were transported to the apical plasma membrane also became insoluble in Triton X-100 but soluble in octylglucoside after extraction from MDCK cells during exocytic transport. These data indicate that the transmembrane domain of NA provides the determinant(s) both for apical transport and for association with Triton X-100-insoluble lipids. PMID- 8709292 TI - Characterization of tumor cell lines derived from murine gammaherpesvirus-68 infected mice. AB - Cell lines were derived from mice with murine gammaherpesvirus-68 (MHV-68) associated lymphoproliferative disease. Four were of an ambiguous phenotype and were MHV-68 negative. One, S11, was a B lymphocyte that contained MHV-68 genomes in both linear and episomal forms and released virus. The line was clonable and grew into tumors in nude mice. This is the first naturally occurring MHV-68 positive B-cell line to be generated, and it will be an invaluable tool for the study of MHV-68 latency. PMID- 8709293 TI - Detection of herpes simplex virus type 1 latency-associated transcript expression in trigeminal ganglia by in situ reverse transcriptase PCR. AB - One of the defining characteristics of herpes simplex virus type 1 (HSV-1) infection is the ability of the virus to establish a lifelong latent state in neurons. We previously demonstrated (R. Ramakrishnan, A.J. Fink, G. Jiang, P. Desai, J. C. Glorioso, and M. Levine, J. Virol. 68:1864-1873, 1994) by in situ PCR that many more neurons contain viral genomes than are detected by in situ hybridization for HSV latency-associated transcripts (LATs). To determine whether all cells which contain genomes express LATs, we examined trigeminal ganglia for LATs 1 and 8 weeks after corneal scarification with ribonucleotide reductase deficient HSV-1 by in situ reverse transcriptase PCR. The number of LAT-positive cells detected by in situ reverse transcriptase was substantially greater than the number of cells positive by in situ hybridization and appeared to be similar to the number of cells containing HSV genomes by in situ PCR and the number of ganglionic neurons that project to the cornea as detected by retrograde labeling with Fluorogold. These results demonstrate LAT expression in many neurons containing HSV-1 genomes. PMID- 8709294 TI - In vitro establishment of lytic and nonproductive infection by herpes simplex virus type 1 in three-dimensional keratinocyte culture. AB - The F strain of herpes simplex virus type 1 (HSV-1) was tested for its ability to produce lytic or nonproductive infection in squamous epithelial cells cultured in a three-dimensional organotypic tissue culture. For the tissue culture, we used HaCat cells (immortalized skin keratinocytes) and normal fibroblasts derived from the skin. The cultures were infected with HSV-1 (5 PFU) either when the epithelial cells had grown as a monolayer with a confluence of 80% on the collagen fibroblast gel or 30 min after lifting of the epithelial cells into the air-liquid interface. The cultures were collected 1 week after inoculation. Typical cytopathic effects of HSV infection (ballooning and reticular degeneration with multinucleate giant cells) were seen only in those cultures in which the epithelial cells were infected before lifting. The presence of HSV was confirmed by DNA and RNA in situ hybridization and PCR. No morphological changes were found in cultures infected after lifting into the air-liquid interface. No infectious virus was recovered either from cells or culture supernatant. However, these cultures were positive for HSV DNA on PCR and showed expression of the LAT gene by in situ hybridization and Northern blot (RNA) hybridization. The present results indicate that both nonproductive and lytic HSV infection can be produced in vitro and the outcome of the infection depends on the time of viral inoculation in relation to epithelial maturation. PMID- 8709295 TI - Intracytoplasmic sperm injection: a treatment for extreme oligospermia. AB - PURPOSE: We evaluated the efficacy of intracytoplasmic sperm injection in patients with extreme oligospermia. MATERIALS AND METHODS: A total of 67 intracytoplasmic sperm injection cycles was attempted in 58 infertile couples in which the husbands had extreme oligospermia (less than 100,000 spermatozoa per ml. ejaculate). RESULTS: Fertilization was achieved in 65 of 67 cycles. Mean fertilization rate per cycle was 66.4%. A total of 18 clinical pregnancies was obtained, for a pregnancy rate of 26.8% per started cycle. There were 4 miscarriages and 8 live births from 5 deliveries. Nine pregnancies are ongoing. CONCLUSIONS: Intracytoplasmic sperm injection in patients with extreme oligospermia is associated with high fertilization rates and offers the chance of pregnancy to these otherwise intractably infertile couples. PMID- 8709296 TI - Seminal vesicle aspiration of fertile men. AB - PURPOSE: Whether sperm normally reside in the seminal vesicles of fertile men without ejaculatory duct obstruction, and the effect of duration of sexual abstinence on results of seminal vesicle aspiration were determined. MATERIALS AND METHODS: Bilateral seminal vesicle aspiration was performed on 12 fertile volunteers under transrectal ultrasound guidance with randomization according to 0 and 5 days of abstinence. Seminal vesicle aspirates were examined microscopically for number and motility of sperm. A positive aspirate was defined as greater than 3 sperm per high power microscopic field. RESULTS: Duration of abstinence had a significant effect on outcome of seminal vesicle aspiration in fertile volunteers. No volunteer with 0 days of abstinence had a positive aspirate from either seminal vesicle, whereas a third of those with 5 days of abstinence had at least 1 positive seminal vesicle aspirate. CONCLUSIONS: Significant numbers of sperm are not normally found in the seminal vesicles of fertile men immediately after ejaculation. Diagnostic seminal vesicle aspiration should be performed after 0 days of abstinence. Longer periods of sexual abstinence may be used in patients undergoing seminal vesicle harvesting of sperm for assisted reproduction. PMID- 8709297 TI - Sperm quality improvement in cryopreserved human semen. AB - PURPOSE: We determined if separation of spermatozoa (washed) on a discontinuous colloidal suspension of silica (Percoll) density gradient before cryopreservation improves post-thaw motility compared to an unprocessed (raw) cryopreserved sample. MATERIALS AND METHODS: Ten normal healthy volunteers recruited into the andrology laboratory donor program were studied. Raw and washed cryopreserved spermatozoa were compared for loss of motility with time, motion characteristics, viability and membrane integrity after incubation for 1, 6 and 24 hours. Within group comparisons were made to baseline measurements (0 hours before incubation). RESULTS: Raw and washed cryopreserved spermatozoa showed statistically significant decreases in motility and other motion characteristics after thawing. There were significant decreases in motility and other motion characteristics after incubation periods of 1, 6 and 24 hours, and significant decreases in viability and membrane integrity at 6 and 24 hours in the unprocessed spermatozoa. Although, motility and motion characteristics of washed samples decreased significantly with longer incubation periods, loss of motility with time (longevity) was greater in raw samples. Washed samples retained greater sperm motility for up to 24 hours (p < 0.03). CONCLUSIONS: Specimens prepared by Percoll separation techniques before freezing offer the possibility of selecting spermatozoa that retain motility for up to 24 hours. This finding can be of benefit for couples undergoing intrauterine insemination to achieve pregnancy. PMID- 8709298 TI - Infertility. PMID- 8709299 TI - Quantification of voiding dysfunction in patients awaiting transurethral prostatectomy. AB - PURPOSE: We investigated the role of advanced urodynamic analysis in the diagnostic evaluation for transurethral resection of the prostate. MATERIALS AND METHODS: A total of 62 consecutive patients awaiting transurethral resection of the prostate with the American Urological Association symptom score of more than 12, peak flow rate less than 13 ml. per second and significant residual urine underwent routine video pressureflow studies with advanced urodynamic analysis to quantify outflow function. Postoperative outcome was assessed at 3 months by symptom scores, uroflowmetry and ultrasonography (residual urine). RESULTS: The parameters used in the diagnostic evaluation for transurethral resection of the prostate did not correlate with urodynamic diagnosis of unequivocal obstruction. Weak detrusor contractility (without significant obstruction) accounted for voiding dysfunction in 50% of the patients. The results of transurethral resection of the prostate were significantly better in patients with unequivocal compared to equivocal obstruction. Furthermore, patients with unequivocal obstruction but weak detrusor contractility had a significantly poorer treatment outcome. CONCLUSIONS: Advanced urodynamic analysis in the diagnostic evaluation of patients with symptomatic benign prostatic hyperplasia can predict treatment outcome and, therefore, allows the urologist to choose the most appropriate therapeutic option for individuals. PMID- 8709300 TI - The correlation between bladder outlet obstruction and lower urinary tract symptoms as measured by the international prostate symptom score. AB - PURPOSE: We studied the relationship between lower urinary tract symptoms as measured by the international prostate symptom score (I-PSS) and urodynamic findings in elderly men. MATERIALS AND METHODS: We evaluated 803 consecutive patients with lower urinary tract symptoms via the I-PSS and urodynamics with pressure-flow studies. RESULTS: A statistically significant correlation was found between all I-PSS questions (except intermittency) and objective parameters of obstruction. However, the clinical significance of this finding is minimal because a large overlap of symptom scores exists among patients with different grades of bladder outlet obstruction. The filling component of the I-PSS correlated somewhat better with obstruction than did the voiding component. CONCLUSIONS: It seems impossible to diagnosis bladder outlet obstruction from symptoms alone. It does not even seem possible to define subgroups in which further urodynamic examination is indicated. PMID- 8709301 TI - Variability of clinical and pressure-flow study variables after 6 months of watchful waiting in patients with lower urinary tract symptoms and benign prostatic enlargement. AB - PURPOSE: We quantified the physiological variability of clinical and pressure flow study variables in patients with symptomatic benign prostatic enlargement. MATERIALS AND METHODS: Symptom scores were measured, and advanced urodynamic studies with pressure-flow analysis were performed in 178 patients before and 6 months after a period a watchful waiting. RESULTS: Patients without bladder outlet obstruction experienced significant symptomatic improvement. Symptoms in patients with obvious bladder outlet obstruction did not improve significantly. The reproducibility of mean pressure-flow variables was evident. However, there was an important intra-individual variability. Patients with obvious bladder outlet obstruction showed a significant decreases in detrusor pressure at maximal flow of 14cm. water, a significant decrease in the urethral resistance factor of 7 cm. water and a significant decrease of 1 obstruction class on the linear passive urethral resistance relation nomogram, indicating less severe bladder outlet obstruction. CONCLUSIONS: Mean differences among therapy groups must be regarded critically, especially when the difference are slight and possibly within physiological variability. PMID- 8709302 TI - Serum prostate specific antigen levels after transurethral resection of prostate: a longitudinal characterization in men with benign prostatic hyperplasia. AB - PURPOSE: We studied the long-term effects of adenomectomy on serum levels of prostate specific antigen (PSA) in men with benign prostatic hyperplasia (BPH). MATERIALS AND METHODS: A consecutive series of 82 men undergoing adenomectomy for BPH between 1990 and 1992 was studied. PSA levels were determined before and serially after operation for as long as 5 years. RESULTS: Mean PSA decreased from 4.6 ng/ml. preoperatively to 0.7 ng./ml. 6 months postoperatively. These low levels were maintained throughout the 5-year observation period. The PSA decrease correlated with grams of tissue removed (r = 0.54, p < 0.001) and averaged 0.11 ng./ml./gm. Postoperatively mean PSA velocity was 0.01 ng./ml. per year, that is essentially flat, and it was not influenced by patient age, race, type of operation, grams of tissue removed, baseline PSA, PSA density of T1a lesions (7). One to 5 years after adenomectomy 6 of the 82 men had invasive prostatic carcinoma, and PSA levels and velocities remained low. CONCLUSIONS: These data support the concept of transition zone primacy in determining serum PSA. Furthermore, they suggest a possible need for a modified reference range when using PSA to screen for prostatic carcinoma in the estimated 3 million men alive in the United States who previously underwent adenomectomy for BPH. PMID- 8709303 TI - Symptom scores, watchful waiting and prostate specific antigen levels in benign prostatic hyperplasia. PMID- 8709304 TI - Free-to-total prostate specific antigen ratio as a single test for detection of significant stage T1c prostate cancer. AB - PURPOSE: We investigated whether impalpable, invisible (stage T1c) but significant prostate cancer can be detected better by determining the free-to total prostate specific antigen (PSA) ratio of equivocal PSA serum levels. MATERIALS AND METHODS: The specificity of free-to-total PSA ratio using research monoclonal enzyme immunoassays was compared to that of PSA greater than 4.0 ng./ml. in 117 consecutive patients with PSA 3 to 15 ng./ml. (Hybritech Tandem-R assay) due to untreated benign prostatic hypertrophy or prostate cancer. Of the patients 77% underwent adenectomy or radical prostatectomy with thorough pathological evaluation of surgical specimens. RESULTS: Benign prostatic hypertrophy had a greater median free-to-total PSA ratio than stages T1c and T2 or greater prostate cancer (0.16 versus 0.09 and 0.11 ng./ml., p = 0.0001 and p = 0.0268, respectively). In stage T1c prostate cancer, areas under receiver operating characteristic curves were 0.58 and 0.84 for PSA and free-to-toal PSA ratio, and free-to-total PSA ratio correlated with prostate volume (r = 0.49, p = 0.005) and Gleason score (r = -0.37, p = 0.036). Pathologically, 84% of stage T1c cancers were significant and comparable to stage T2 or greater cancers. CONCLUSIONS: Free-to-total PSA ratio enhances the efficacy of PSA measurement by improving specificity for detecting impalpable, invisible but significant stage T1c prostate cancer. PMID- 8709305 TI - Prostatic intraepithelial neoplasia: influence of clinical and pathological data on the detection of prostate cancer. AB - PURPOSE: We attempted to characterize patients diagnosed with prostatic intraepithelial neoplasia without concurrent cancer on biopsy who had prostate cancer on subsequent biopsy. MATERIALS AND METHODS: The records of 93 patients with low and high grade prostatic intraepithelial neoplasia without concurrent cancer on initial biopsy were analyzed. The relationships among prostatic intraepithelial neoplasia grades, patient age, digital rectal examination, serum prostate specific antigen (PSA), transrectal ultrasound appearance and final pathological results were investigated. RESULTS: Subsequent carcinoma was found on repeat biopsy in 13.3% of patients with low grade and 47.9% with high grade prostatic intraepithelial neoplasia (p < 0.001). In the former group digital rectal examination, patient age, serum PSA and transrectal ultrasound were not predictive of cancer. Transrectal ultrasound appearance, digital rectal examination and serum PSA were statistically different between high grade prostatic intraepithelial neoplasia with and without subsequent cancer (p < 0.001, p = 0.008 and p = 0.016, respectively, univariate analysis). On multivariate analysis of patients with high grade prostatic intraepithelial neoplasia only digital rectal examination and PSA were predictive of subsequent carcinoma. CONCLUSIONS: High grade prostatic intraepithelial neoplasia is a strong predictor of subsequent cancer, especially in men with abnormal digital rectal examination and elevated serum PSA. Patients with high grade prostatic intraepithelial neoplasia should undergo repeat biopsy to exclude cancer. Further investigations are needed to optimize the treatment of patients with low grade prostatic intraepithelial neoplasia. PMID- 8709306 TI - Pathological characteristics and prognosis of nonpalpable and palpable prostate cancers with a Hybritech prostate specific antigen of 4 to 10 ng./ml. AB - PURPOSE: We compared the surgical pathological findings and postoperative course of patients with palpable and nonpalpable prostate cancers. MATERIALS AND METHODS: All patients with untreated prostate specific antigen (PSA) 4 to 10 ng./ml. who underwent radical prostatectomy between December 1984 and December 1993 were reviewed to select 61 with clinical stage T1c (nonpalpable) with stages T2a to c (palpable) disease. RESULTS: Nonpalpable cancers were smaller (2.99 versus 4.42 cc for palpable tumors), had smaller volumes of Gleason grade 4 to 5 cancer (0.66 versus 1.32 cc, respectively) and were less likely to have positive surgical margins (13 versus 22%, respectively) or significant (1 cm. or more) capsular penetration (10 versus 26%, respectively). Nonpalpable and palpable cancers had similar rates of seminal vesicle invasion (3.3 versus 4.3%, respectively) and positive lymph nodes (1.6 versus 0%, respectively). More than 90% of patients with nonpalpable cancer were biochemically cancer-free postoperatively, and the remainder were alive with disease after a mean followup of 25.1 months, compared to 69% disease-fee, 28% alive with disease and 2.5% dead of prostate cancer after mean followup of 43.8 months among those with palpable disease. CONCLUSIONS: We conclude that nonpalpable prostate cancers are pathologically more favorable than palpable prostate cancers with PSA 4 to 10 ng./ml. Our preliminary results also indicate that nonpalpable cancers are less likely to recur postoperatively than palpable cancers with a similar PSA range. PMID- 8709307 TI - Distinguishing clinically important from unimportant prostate cancers before treatment: value of systematic biopsies. AB - PURPOSE: We assessed the ability of diagnostic tests to distinguish clinically unimportant cancers. MATERIALS AND METHODS: We correlated T stage (based on digital examination and ultrasound), prostate specific antigen (PSA), PSA density and pathological features of cancer in systematic biopsy specimens with features of cancer in 170 radical prostatectomy specimens. Clinically unimportant cancers were defined as small (0.5 cm.3 or less), well or moderately differentiated and confined to the prostate. RESULTS: Of the patients 10% had an unimportant cancer. On logistic regression analysis the 2 significant predictors were maximum length of cancer in any core and PSA density. Of 12 patients with maximum cancer length 2 mm. or less and PSA density less than 0.1., 75% had an unimportant cancer compared to 5% of the remaining 158 (p < 0.0005). CONCLUSIONS: Quantitative analysis of systematic biopsy specimens combined with PSA density provides valuable staging information and helps to identify cancers of low biological potential. PMID- 8709308 TI - Ki-67 expression is a prognostic marker of prostate cancer recurrence after radical prostatectomy. AB - PURPOSE: We assessed the cellular proliferation of clinically localized prostate cancer by immunohistochemistry using the monoclonal antibody MIB to Ki-67 antigen in an attempt to identify associations between proliferative indexes and disease progression following radical prostatectomy. MATERIALS AND METHODS: Ki-67 proliferative antigen was evaluated using MIB 1 monoclonal antibody in archival paraffin embedded radical prostatectomy specimens from 180 patients followed for 1 to 9 years (mean 4.4). The percentage of tumor nuclei expressing Ki-67 antigen was measured and assigned and MIB 1 score (none or rare--negative, 1+--low score and 2 to 4+--high score) and analyzed for prostate specific antigen, stage, age, race, grade and serological recurrence postoperatively. RESULTS: There was a significant association between MIB 1 score and nuclear grade (p < 0.001), Gleason score (p < 0.001) and pathological stage (p = 0.01). Patients with a high MIB 1 score had earlier progression and a lower 5-year recurrence-free survival rate (44%) than those with negative MIB 1 scores (71%, p < 0.001). In multivariate Cox regression analysis with backward elimination, pathological stage (p < 0.01), pretreatment prostate specific antigen (p = 0.04) and MIB 1 score (p = 0.05) were statistically significant predictors of disease-free survival, and patients with a high MIB 1 score were 3.1 times as likely to have recurrence as those with a negative score. Controlling for stage, patients with organ confined disease and a high MIB 1 score had a lower 5-year disease-free survival rate (68%) than those with a low MIB 1 score (95%, p < 0.01). CONCLUSIONS: Proliferative activity as measured by the Ki-67 proliferative antigen, MIB 1, appears to be a prognostic marker of recurrent prostate cancer after radical prostatectomy. PMID- 8709309 TI - Significant versus insignificant prostate cancer--can we identify the tigers from the pussy cats? PMID- 8709311 TI - Blood loss during radical retropubic prostatectomy: is preoperative autologous blood donation indicated? AB - PURPOSE: We assessed the requirement for transfusion of allogeneic blood in a contemporary series of patients who did not deposit autologous blood before radical retropubic prostatectomy. MATERIALS AND METHODS: After a policy was adopted in which preoperative autologous blood was not donated, 124 consecutive patients underwent radical retropubic prostatectomy. Type and screen for allogeneic blood were routinely available but neither hemodilation nor a cell saver was used. RESULTS: Mean intraoperative blood loss was 579 cc and mean postoperative serum hematocrit was 33%. Only 3 patients (2.4%) required blood products due to intraoperative blood loss (2) and postoperative bleeding from a duodenal ulcer (1). CONCLUSIONS: Transfusion of blood products was required in a small percentage of our patients even without autologous blood donation. Therefore, the overall cost of care is decreased but, more importantly, the potential risks associated with autologous or allogeneic blood transfusion are eliminated. PMID- 8709310 TI - Transitional cell carcinoma involving the prostate with a proposed staging classification for stromal invasion. AB - PURPOSE: We investigated the effect on survival of transitional cell carcinoma of the prostatic urethra, ducts and stroma, and determined the difference between prostatic stromal involvement occurring via direct extension through the bladder wall versus stromal invasion arising intraurethrally. MATERIALS AND METHODS: Between August 1971 and December 1989, 489 men underwent radical cystoprostatectomy for transitional cell carcinoma, including 143 (29.2%) identified with prostate involvement by transitional cell carcinoma, in the cystectomy specimen. Patients were separated into 2 groups: 1-19 in whom the primary bladder tumor extended full thickness through the bladder wall to invade the prostate (classified as P4a) and 2-124 in whom prostate involvement arose from within the prostatic urethra. RESULTS: Five-year recurrence-free and overall survival rates were 25 and 21%, respectively, in group 1 versus 64 and 55%, respectively, in group 2. In the 124 patients in group 2 survival rates were similar for those with prostatic urethral tumors or carcinoma in situ and ductal tumors (no stromal invasion). Five-year overall survival rates without and with stromal invasion were 71 and 36%, respectively (p < 0.0001). Transitional cell carcinoma of the prostatic urethra or ducts does not alter survival predicted by primary bladder stage alone. Prostatic stromal invasion arising intraurethrally significantly decreases survival, which varies based on primary bladder stage (64.6% in stage P1, 30.8% in stages P2/P3a and 13.6% in stage P3b, p = 0.0001). P1 bladder tumors with prostatic stromal invasion arising intraurethrally had a significantly higher survival rate than P4a tumors (64.6 versus 21%, p = 0.0001). P3b bladder tumors with stromal invasion had a survival rate similar to that of P4a tumors (p = 0.78). CONCLUSIONS: Prostatic urethral or ductal transitional cell carcinoma does not alter survival determined by primary bladder stage alone and it should not be classified as P4a. Prostatic stromal involvement arising intraurethrally significantly decreases survival predicted by primary bladder stage alone. P1 bladder tumors with prostatic stromal invasion arising intraurethrally have a significantly higher survival rate than P4a tumors and they should be separately classified as P1str. Muscle invasive (P2/P3a) bladder tumors with stromal invasion have a higher survival rate than P4a tumors (no statistical significance) and they should be designated separately (that is P2str). P3b bladder tumors with prostatic stromal invasion arising intraurethrally are indistinguishable from P4a tumors. PMID- 8709312 TI - Radical prostatectomy: is complete resection of the seminal vesicles really necessary? AB - PURPOSE: We determined the frequency of prostate cancer extension into the distal 1 cm. of seminal vesicles, and reconsidered whether complete excision of the seminal vesicles during radical prostatectomy is always necessary. MATERIAL AND METHODS: After en bloc removal with the specimen in 71 consecutive radical prostatectomies, the distal 1 cm. of each seminal vesicle was transected and separately analyzed for tumor involvement. RESULTS: Mean patient age was 61.8 years (range 40 to 72). Preoperative prostate specific antigen (PSA) ranged from 0.8 to 37 ng./dl. (median 7.3), and 18 patients had a PSA of 10 or more. Clinical stages were T1b in 1 case T1c in 37, T2a in 12, T2b in 10, T2c in 6 and T3a in 1. Preoperative Gleason sums ranged from 4 to 8 (median 6) with 21 patients (30%) having a sum of 7 or more. Of 71 patients 12 (17%) and seminal vesicle invasion (5 bilaterally). In no case did tumor extend into the distal 1 cm. of the seminal vesicle. PSA at diagnosis ranged from 4.2 to 30 ng./dl., with 4 of 12 patients having a PSA of 10 or more. Preoperative clinical stages were T1c in 5 cases, T2a in 3, T2b in 2 and T2c in 2. Five of the 12 patients (42%) had positive surgical margins and 11 (92%) had a postoperative Gleason sum of 7 or more. CONCLUSIONS: In 71 consecutive patients undergoing radical prostatectomy no tumor was found in the distal 1 cm. of the seminal vesicles, including 12 with seminal vesicle invasion. We continue to advocate complete excision of the seminal vesicles during radical prostatectomy. However, if dissection is difficult and a small fragment is left behind, the prognosis is unlikely to be altered. PMID- 8709314 TI - Prostate cancer and aspects of radical prostatectomy. PMID- 8709313 TI - Mortality in prostate cancer. AB - PURPOSE: We evaluated in patients with prostate cancer whether treatment is associated with in decreased mortality and whether the association of treatment with mortality varies with calendar time. MATERIALS AND METHODS: Using the 146,979 prostate cancer patients from the 1973 to 1990 public use tape of the Surveillance, Epidemiology and End Results program, we performed survival analysis and multivariate proportional hazards modeling to estimate the relative risk of disease specific and overall mortality. RESULTS: In men with prostate cancer, advanced age, black race, high tumor stage and not having treatment were independently associated with disease specific and overall mortality. The relative risk of mortality in treated patients decreased significantly from 1973 to 1990. CONCLUSIONS: The fact that advanced age and black race are associated with disease specific mortality (even when treatment and stage are controlled) is a new observation, which suggests that tumor biology and/or response to treatment is worse in elderly and black men. Overall, the data are consistent with the hypothesis that treatment of prostate cancer is associated with lower disease specific and overall mortality rates. The decreases in relative mortality in treated patients from 1973 to 1990 indirectly support the theory that changes in patterns of care from 1973 to 1990 had a beneficial effect on mortality. This observation implies that future studies should account for calendar time when interpreting outcomes data. PMID- 8709315 TI - Acucise endopyelotomy: assessment of long-term durability. AB - PURPOSE: We evaluated the long-term efficacy of Acucise endopyelotomy. MATERIALS AND METHODS: A total of 28 patients with 28 ureteropelvic junction obstructions was reevaluated 2 or more years after Acucise endopyelotomy (mean 32.5 months). Subjective analysis was done with analog pain scales and objective analysis was performed with diuretic renal scintigraphy. RESULTS: Subjective followup was available for all 28 patients, and 17 (61%) had a favorable response with 36% totally free of pain and 25% markedly improved. Among 26 patients with objective followup (93%) 21 (81%) had a patient ureteropelvic junction based on a diuretic renal scan with a half-time of less than 10 minutes or a normal Whitaker test. Among all regularly followed patients failure occurred uniformly within 1 year. CONCLUSIONS: Acucise endopyelotomy is an effective and durable method for treating ureteropelvic junction obstruction. PMID- 8709316 TI - Endourological management of ureteral obstruction after renal transplantation. AB - PURPOSE: We evaluated endourological treatment of ureteral obstruction after renal transplantation. MATERIALS AND METHODS: Between January 1986 and December 1993, 582 kidney transplantations were performed at our center, and ureteral obstruction was suspected in 31 cases (5.3%). RESULTS: Initial treatment consisted of retrograde placement of an internal stent in 6 patients and percutaneous nephrostomy in 25. Due to upper tract dilatation obstruction could not be diagnosed in 3 patients, and rejection was the cause of decreasing renal function. Obstruction was temporary in 8 of the remaining 28 patients, including 6 in whom a Double-J stent was introduced in a retrograde manner without anesthesia. In the other 2 patients was well as the 20 with definitive obstruction, cannulation of the transplant orifice without anesthesia was unsuccessful and percutaneous nephrostomy drainage was necessary. Even with general anesthesia a guide wire could not be passed along the stricture in a retrograde or antegrade fashion in 7 of the 20 patients with definitive obstruction and open surgery was performed. The remaining 13 patients underwent dilation with (9) or without (4) diathermic incision. All 4 patients treated with dilation only had recurrent obstruction, while 9 treated with dilation and incision had no recurrence after a minimum followup of 27 months (mean 58). CONCLUSIONS: Modern endourological procedures have replaced open reconstructive surgery in the majority of patients with ureteral obstruction after renal transplantation. PMID- 8709317 TI - Laparoscopic nephrectomy via the retroperitoneal approach. AB - PURPOSE: We evaluated efficacy of the retroperitoneal approach for laparoscopic nephrectomy of kidneys with benign disease. MATERIALS AND METHODS: Eight men and 12 women (mean age 55 years) with severely damaged kidneys underwent laparoscopic retroperitoneal nephrectomy. One patient had a history of multiple open abdominal and gynecological operations. Kidneys were removed laparoscopically from the working space, which was created by finger and balloon dissection, and maintained by carbon dioxide insufflation in the retroperitoneal cavity. RESULTS: All kidneys were removed successfully via this procedure. Mean operative time was 3.3 hours and mean estimated blood loss was 135 ml. One patient experienced bleeding from the injured capsular artery just after removal of the kidney. CONCLUSIONS: The retroperitoneal approach is recommended for laparoscopic nephrectomy. PMID- 8709318 TI - The gasless laparoscopic Burch bladder neck suspension: early experience. AB - PURPOSE: We evaluated the gasless extraperitoneal laparoscopic Burch bladder neck suspension. MATERIALS AND METHODS: This retrospective study included 47 patients with type II stress urinary incontinence treated sequentially with this technique between September 1994 and September 1995. Balloon dissection was used to develop the extraperitoneal space. A mechanical retraction system was used with conventional laparotomy instruments to perform laparoscopic Burch bladder neck suspension. RESULTS: Of the 47 patients who underwent this procedure 3 (6%) required conversion to an open operation. Obesity and previous pelvic surgery were not contraindications to this technique. The only major complication involved blood loss necessitating conversion to an open operation. Average operative time was 96.4 minutes and average hospital stay was 3.5 days. Followup at 2 to 15 months (mean 8.2) indicated successful results (that is no pads were required) in 44 patients (90%). CONCLUSIONS: Gasless laparoscopic bladder neck suspension has a lower open surgery rate, and may be performed more rapidly than conventional carbon dioxide laparoscopic Burch bladder neck suspension. Previous multiple operations and obesity are not contraindications to the technique. PMID- 8709319 TI - Laparoscopic retroperitoneal lymph node dissection for nonseminomatous germ cell tumors: indications and limitations. AB - PURPOSE: We describe our experience with laparoscopic retroperitoneal lymph node dissection in 26 patients with nonseminomatous germ cell tumors: 17 had stage I disease with no clinical (computerized tomography, ultrasound or tumor markers) evidence of metastases and 9 (2 with stage IIb and 7 with stage IIc disease) had residual tumor after chemotherapy but with negative tumor markers. Laparoscopic dissection was performed to assess more fully pathological status of the relevant retroperitoneal lymph nodes in both groups. MATERIALS AND METHODS: The patient was positioned and trocars were introduced at sites similar to that used for transperitoneal laparoscopic nephrectomy (flank position with 3, 10 mm. and 2, 5 mm. ports). After the white line of Toldt was incised and the colon was reflected anteromedially, the retroperitoneal space was exposed. The landmarks of lymph node dissection were then isolated, including the ureter, aorta, inferior vena cava and both renal veins. Lymph node dissection was performed identical to that for open surgery, with a modified template including the paracaval, interaortocaval, upper preaortic and right common iliac nodes for right tumors, and para-aortic and upper preaortic nodes for left tumors. Lymph node chains were retrieved with a small organ bag. RESULTS: The procedure was completed successfully in 16 of 17 patients with stage I disease (mean duration 268 minutes for the left and 312 minutes for the right sides). No intraoperative complications were encountered. One patient had delayed ureteral stenosis requiring operative repair, 1 had a pulmonary embolism with an uneventful outcome and 1 who underwent laparoscopic retroperitoneal lymph node dissection on the right side later had retrograde ejaculation. Embryonal carcinoma was found in 1 of the 17 patients. Average postoperative hospital stay was 4.5 days for patients without complications or conversion to an open procedure. After a median followup of 27 months no patient had regional relapse but 2 had pulmonary metastases that were treated successfully with 3 cycles of platinum based chemotherapy. Laparoscopic dissection was significantly more difficult in patients with stage II tumors after chemotherapy. Only in 2 patients with stage IIb disease was laparoscopic lymphadenectomy successful. In 5 of the 7 patients with stage IIc cancer portions of the dissection had to be done after conversion to an open (conventional) operation via a small incision (suprainguinal or pararectal). In 1 patient the laparoscopic approach was completely abandoned and converted to an open operation via a standard midline incision. In all 9 cases histopathological examination revealed complete necrosis. No patient has evidence of disease. CONCLUSIONS: Our preliminary experience suggests that a modified laparoscopic retroperitoneal lymph node dissection is feasible for stage I tumors. However, it cannot be recommended after previous chemotherapy (stages IIb and IIc disease). PMID- 8709320 TI - Endourology. PMID- 8709321 TI - Trimming a tether: a simple way to prevent complications related to tethered ureteral stents. AB - PURPOSE: A method to prevent complications related to a tethered ureteral stent was assessed. MATERIALS AND METHODS: In 17 male patients tethers were trimmed just short of the meatus after insertion of a ureteral stent. At stent removal a rigid cystoscope was inserted into the anterior urethra, and the edge of the tether was grasped with foreign body forceps. RESULTS: Only 1 patient required removal of the tether due to severe urethral discomfort. Pain at cystoscopy for removal of the ureteral stent was minimal. CONCLUSIONS: This method is useful for improving the quality of life during the catheterization period without decreasing effectiveness of a conventional ureteral stent with a tether. PMID- 8709322 TI - Excisional plication of the ileocecal valve: a useful adjunct for the construction of continent urinary diversions. AB - PURPOSE: A variety of techniques exist to reinforce the ileocecal valve for use as a nonrefluxing mechanism in continent urinary diversion. We report short-term and long-term followup for a new technique of excisional plication of the valve. MATERIALS AND METHODS: Since 1988 we performed 16 procedures in 15 patients. The technique was used for repair of an incompetent ileocecal valve to restore continence in 2 patients in whom continent cutaneous diversion failed, and to correct a massively refluxing ileocecal valve following previous augmentation with an ileocecal segment in 1. The remaining patients underwent the procedure in conjunction with continent cutaneous ileocecal diversion with a modified Indiana pouch. RESULTS: To date no patient undergoing continent cutaneous diversion required reoperation to correct incontinence. Two patients wear a protective pad during the day, while the remainder have been completely dry during followup. CONCLUSIONS: Excisional plication is a versatile and simple procedure that may be used to assist construction and repair of ileocecal valves in conjunction with continent urinary diversion. PMID- 8709323 TI - Access technique for retroperitoneoscopy. AB - PURPOSE: The access technique for retroperitoneoscopy is not well established, and differs from transperitoneal laparoscopic access in 3 key aspects: 1) location and technique of primary trocar placement, 2) optimal positioning of the balloon dilator and 3) technique for safe placement of secondary ports. Our method of obtaining retroperitoneoscopic access addresses these issues. MATERIALS AND METHODS: A total of 37 patients underwent retroperitoneoscopic surgery of the kidney and upper ureter. RESULTS: Our technique facilitates balloon placement within Gerota's fascia, minimizes peritoneal injury and optimizes port placement during retroperitoneoscopic surgery. CONCLUSIONS: Although our success rate for placing the balloon within Gerota's fascia has improved, additional experience is required to achieve subfascial balloon placement more consistently. PMID- 8709324 TI - The effect of urapidil on neurogenic bladder: a placebo controlled double-blind study. AB - PURPOSE: We assessed the efficacy and safety of urapidil, a new alpha-blocker, for the treatment of neurogenic bladder dysfunction in a prospective double-blind trial. MATERIALS AND METHODS: A total of 136 patients was randomly assigned to receive placebo (group 1), or treatment with 30 mg. of urapidil for 4 weeks (group 2) or 30 mg. for 2 weeks followed by 60 mg. for 2 weeks (group 3). RESULTS: Urinary frequencies decreased significantly only in group 3. The sum of obstructive symptom scores decreased significantly in all groups. However, a significant difference in subjective symptoms was not noted among the groups. Average and maximum flow rates, and residual urine improved significantly in groups 2 and 3 but not in group 1. Regarding cystometric parameters, no significant changes were noted in any group. In the pressure-flow study, the pressure at maximum flow rate and minimum urethral resistance (pressure at maximum flow rate divided by the square of maximum flow rate) decreased significantly only in group 3. In the urodynamic assessment, significant improvement was noted in groups 3 versus 1. There were no significant differences between the sexes or among underlying diseases with regard to symptomatic and urodynamic improvements. Side effects were noted in 2 patients in group 3 and 1 in group 2, none of which was severe. CONCLUSIONS: Urapidil improved voiding dysfunction in patients with a neurogenic bladder and decreased urethral resistance in a dose related fashion. PMID- 8709325 TI - Urodynamic assessment of urethral sphincter function in post-prostatectomy incontinence. AB - PURPOSE: Direct measurement of maximum urethral pressure by urethral profilometry has been used widely to assess urethral sphincter function. We attempted to determine if there was any relationship between maximum urethral pressure, which is measured at the level of the membranous urethra, or extrinsic urethral sphincter function, and the amount of abdominal pressure needed to cause leakage (abdominal leak point pressure) in men with post-prostatectomy incontinence. We also examined the relationship between external sphincter function and continence or incontinence. MATERIALS AND METHODS: We retrospectively evaluated fluoro urodynamics performed in 37 men with post-prostatectomy incontinence. Urodynamic study consisted of measurement of maximum urethral and abdominal leak point pressures, and assessment of extrinsic sphincter function by pressure measurements and radiographically. RESULTS: Data were analyzed on 27 patients for whom abdominal leak point and maximum urethral pressures were available. Mean maximum urethral pressure was 52.5 cm. water (range 20 to 165) and mean abdominal leak point pressure was 77.8 cm. water (range 27 to 132). Regression analysis was performed between maximum urethral and abdominal leak point pressures. A Pearson correlation coefficient of 0.13834 was calculated (p = 0.4914) indicating virtually no correlation between the 2 measurements in our sample. Extrinsic urethral sphincter was normal in all patients. Only 1 of 37 patients had no evidence of intrinsic sphincter deficiency, that is there was no urine leakage with increases in abdominal pressure and the patient was incontinent solely based on bladder dysfunction (detrusor instability). CONCLUSIONS: Our study indicates that incontinence after prostatectomy due to an increase in abdominal pressure (stress incontinence) does not depend on extrinsic sphincter function and is not related to maximal urethral pressure. We conclude that post-prostatectomy incontinence due to sphincter dysfunction results from intrinsic sphincter deficiency. In our experience bladder dysfunction is rarely the sole cause of post-prostatectomy incontinence. PMID- 8709326 TI - Re: Adrenalectomy via the dorsal approach: a benchmark for laparoscopic adrenalectomy. PMID- 8709327 TI - Re: Fibrous coagulum: a new cause of retained fragments following extracorporeal shock wave lithotripsy. PMID- 8709328 TI - Re: Subcutaneous urinary diversions for palliative treatment of pelvic malignancies. PMID- 8709329 TI - Re: Ureteroscopic removal of mid and proximal ureteral calculi. PMID- 8709330 TI - Re: Results of a multicenter trial using the BTA test to monitor for and diagnose recurrent bladder cancer. PMID- 8709331 TI - Re: Editorial: what to expect from prostate cancer. PMID- 8709332 TI - Re: Editorial: what to expect from prostate cancer. PMID- 8709333 TI - Re: Measurement of pudendal evoked potentials during feminizing genitoplasty: technique and applications. PMID- 8709334 TI - 10-year experience with prenatal intervention for hydronephrosis. AB - PURPOSE: We evaluated clinical outcomes after attempted prenatal intervention in fetuses with hydronephrosis. MATERIALS AND METHODS: We retrospectively reviewed the histories of 10 fetuses considered for prenatal intervention between 1984 and 1993. One female and 8 male fetuses had bilateral hydroureteronephrosis and profound or progressive oligohydramnios, and 1 male fetus had massive progressive unilateral hydronephrosis. RESULTS: Shunt placement was not recommended and it was refused by the parents in 1 case each. Amniotic shunting was attempted in 8 fetuses between 22 and 28 weeks of gestation. Shunts were successfully placed with decreased hydronephrosis and increased amniotic fluid volume in 4 cases, while shunt placement was not technically possible in the remainder. Two shunts that retracted intra-abdominally at birth required laparotomy for retrieval. Postnatally all patients with shunts had compromised renal function. Of the 4 patients in whom attempts were unsuccessful 3 had mildly diminished renal function and 1 died of nonrenal causes (intraabdominal sepsis) on day 16 of life. No patient with a functioning shunt had postnatal pulmonary problems, whereas 3 without successful intervention had mild respiratory compromise. CONCLUSIONS: No definite advantage was noted in the small number of fetuses that underwent successful shunting. Successful shunt placement did not prevent renal insufficiency. The relief of oligohydraminos may benefit pulmonary function in some patients. The high technical failure and complication rates of in utero intervention should be considered before proceeding. PMID- 8709335 TI - Colonic conduit in children: protection of the upper urinary tract 16 years later? AB - PURPOSE: Complication rates of up to 86.6% have been reported after creation of an ileal conduit. In contrast to others, we construct an antirefluxing isoperistaltic colonic for incontinent diversion in children. MATERIALS AND METHODS: Between 1968 and 1989 a colonic conduit was created in 105 patients up to age 20 years for neurogenic bladder (76), bladder exstrophy (16) and other reasons (13). Long-term followup (minimum 5 years, mean 16.3 years, range 5 to 26) was possible in 84 patients (159 renal units). RESULTS: Early and late stenosis at the ureterocolic anastomosis, and stomal stenosis occurred in 7.6 and 15.5% of the cases, respectively. Renal calculi developed in 8.2% of the renal units. Eight kidneys without function after recurrent pyelonephritis, calculi or stenosis at the ureterocolic anastomosis were removed during followup. Compared to preoperative status 3 of the remaining 151 renal units had increased dilatation of the upper urinary tract at the last followup. A total of 31 patients had complications during followup and 18 underwent conversion to continent urinary diversion. CONCLUSIONS: When incontinent urinary diversion is necessary in children, a colonic conduit with the option of continent conversion should be created before deterioration of the upper urinary tract. PMID- 8709336 TI - Progressive ureteral dilation for subsequent ureterocystoplasty. AB - PURPOSE: The complications associated with bladder augmentation using the gastrointestinal tract are well known. A bladder based ureteral flap is a viable alternative for augmentation. However, its use is limited to cases in which a dilated ureter is already present due to associated pathological conditions, such as reflux. We designed a system for performing progressive dilation in a normal ureter, which can subsequently be used for augmentation. MATERIALS AND METHODS: Ten rabbits underwent unilateral ureteral ligation at the ureterovesical junction and ipsilateral nephrectomy. A silicone catheter was threaded into the proximal ipsilateral ureter and connected to an injection port, which was secured subcutaneously. Ten days to 2 weeks after surgery a saline-antibiotic solution was injected subcutaneously daily into the injection port. Cystography and cystometrography were performed with an accurate measurement of bladder capacity. Ureterography was done initially and then weekly by injecting contrast medium into the ureter. After ureteral dilation ureterocystoplasty was performed in each animal. RESULTS: After 1 month of daily saline-antibiotic solution injections the ureteral units were dilated at least 10-fold, as measured by radiography. The dilated ureteral diameter exceeded that of adjacent colon in each instance. Augmentation cystoplasty was performed with the reconfigured dilated ureteral segment. Repeat cystography and cystometrography showed an average 260% increase in bladder capacity (range 190 to 380%). CONCLUSIONS: Ureteral dilation can be accomplished using our system. At least a 10-fold increase in ureteral size is possible. The dilated ureteral segment can be used effectively for bladder augmentation, avoiding the use of bowel segments. PMID- 8709337 TI - Future directions in lower urinary tract reconstruction. PMID- 8709338 TI - The unintubated ureterotomy endourologically revisited. AB - PURPOSE: The duration of stenting after endoureterotomy is a controversial issue. An even more basic question, however, is whether stent placement is needed at all. We performed a porcine study to address this question. METHODS: A unilateral midureteral stricture was created in 20 minipigs. Six weeks later, 15 pigs underwent endoureterotomy. In 10 animals, a 7F ureteral stent was placed for 1 week. Five pigs remained unstented. Three months later, all 20 ureters were studied radiographically and were harvested. RESULTS: Three of 5 control animals and 2 of 10 stented animals has strictures. In the unstented study group, all 5 animals had a nonobstructing dilation at the site of endoureterotomy. This was corroborated by a normal furosemide washout renal scan in all 3 animals so tested. CONCLUSIONS: The result of unstented endoureterotomy in the porcine model suggests that, after endoureterotomy of secondary midureteral strictures, stenting may be unnecessary. PMID- 8709339 TI - Desmopressin and vasopressin increase locomotor activity in the rat via a central mechanism: implications for nocturnal enuresis. AB - PURPOSE: Nocturnal enuresis is characterized by nocturnal urine volumes exceeding bladder capacity and by inability to wake up to the stimulus of a full bladder. Desmopressin (DDAVP) is believed to be efficient in treating nocturnal enuresis by reducing nocturnal urine production. However, clinical observations indicate an additional mode of action since the drug appears to modify sleep architecture, apparently improving the patient's ability to awaken to the stimulus of a full bladder. Because of this, a possible arousing effect of DDAVP was studied. MATERIALS AND METHODS: The tentative ability of DDAVP and the endogenous hormone vasopressin (AVP) to produce locomotor stimulation in resting rats after both intracerebroventricular and subcutaneous administration was used as an animal model of arousal. In addition brain monoamine biochemistry was analyzed. RESULTS: The intracerebroventricular injection of AVP (0.1 and 1 microgram.) and the intracerebroventricular (0.1, 1, 10 and 100 microgram.) and subcutaneous (90 and 180 microgram.) injections of DDAVP were both associated with a significant increase in the locomotor activity of the animals compared with controls. The biochemical analysis of cerebral monoamines indicated that DDAVP lowers brain dopamine levels after both types of administration. CONCLUSIONS: These results suggest that DDAVP exerts a stimulatory effect in the CNS, which is also observed after peripheral administration. There are also indications for an increase in central dopamine turnover which could explain the registered increase in locomotor activity. PMID- 8709340 TI - Prostanoid production in rabbit corpus cavernosum. II. Inhibition by oxidative stress. AB - PURPOSE: To investigate the effects of reoxygenation following hypoxia on prostanoid production in rabbit penile corpus cavernosum tissue (RCC) in organ culture. MATERIALS AND METHODS: Strips of RCC were incubated in organ culture media under either 21% O2 (control, PO2 approximately 167 mm.Hg) or 0% O2 (hypoxia, PO2 approximately 27 mm.Hg) followed by a reoxygenation period with 21% O2, in the presence or absence of exogenous arachidonate, Tiron or catalase. Prostanoids were measured in collected media by radioimmunoassay. Malondialdehyde levels were measured in RCC following exposure to either control or hypoxia reoxygenation conditions. RESULTS: Under hypoxic conditions, basal release of prostanoids (PGI2, PGF2 alpha, PGE2 and TXB2) was inhibited. Although this inhibition was reversible upon reoxygenation, the recovery was delayed, requiring at least 2 hours of exposure to 21% oxygen to reestablish prostanoid production. Reoxygenation also caused lipid peroxidation as measured by an increase in malondialdehyde levels. When reoxygenation was done in the presence of exogenous arachidonate, recovery of PGI2 production was complete by 1 hour. Reoxygenation in the presence of a scavenger of reactive oxygen species (Tiron) or catalase significantly improved the recovery rate of PGI2 production. CONCLUSIONS: These results show that reoxygenation of hypoxic tissue generates oxidative stress that interferes with the recovery of prostanoid production by alteration of a biosynthetic point(s) upstream from prostaglandin H synthase (PGHS) including, at least, phospholipid peroxidation. PMID- 8709341 TI - The mitotoxin, basic fibroblast growth factor-saporin, effectively targets human prostatic carcinoma in an animal model. AB - PURPOSE: The antitumor activity of the mitotoxin basic fibroblast growth factor saporin (bFGF-SAP) against human prostatic carcinoma DU 145 was examined in athymic nude mice. Therapeutic efficacy was evaluated on the basis of dose, route of administration and treatment schedule. MATERIALS AND METHODS: Chemical conjugate or recombinant bFGF-SAP (0.02 to 50 micrograms/kg.) was administered by intravenous tail injection, intraperitoneal injection, or local or distal subcutaneous injection beginning 5 days (or 60 to 121 days for large tumor studies) after subcutaneous implantation of DU 145 cells. Tumor growth was monitored as long as 140 days by external caliper measurements. RESULTS: Recombinant bFGF-SAP, though less cytotoxic than its chemical conjugate form, effectively targeted DU 145 tumors growing as xenografts in nude mice in a dose dependent manner. Antitumor response to treatment by intravenous, intraperitoneal, or distal subcutaneous injection suggested similar bioavailability of the mitotoxin administered by each route; local subcutaneous injection to the tumor site resulted in statistically better antitumor response. Schedules that included at least 1 bFGF-SAP treatment beyond day 23 increased long-term antitumor efficacy independent of total dose. Moreover, recombinant bFGF-SAP induced dramatic reduction of large, established tumors. CONCLUSIONS: These studies suggest a therapeutic potential for bFGF receptor-directed toxins in targeting prostate cancer; further, these data suggest that treatment of established tumors (> 3 weeks) results in qualitatively and quantitatively improved tumor responses. PMID- 8709342 TI - Increased vascular collagen and noncollagenous protein synthesis contributes to sustain chronic phase of two-kidney, one-clip renovascular hypertension. AB - PURPOSE: Although the enhanced renin-angiotensin (R-A) system responsible for two kidney, one-clip (2K-1C) hypertension is well known, there may be a shift with time so that this, hemodynamic factor plays a less important role and increased vascular resistance is predominant in sustaining hypertension. While increased vascular protein synthesis has been demonstrated in genetically hypertensive rats, we evaluated the possible relationship between vascular protein synthesis and 2K-1C hypertension with special reference to the R-A system. MATERIALS AND METHODS: Two-kidney one-clip rats were treated with splanchnicotomy, beta aminopropionitrile (collagen inhibitor), or captopril (angiotensin converting enzyme inhibitor) in the acute or chronic hypertensive phase. 3H-proline was injected into rats, and incorporation rates of 3H-proline into vascular collagen, noncollagenous protein and elastin were counted. The plasma level of the R-A system was assayed. RESULTS: In the acute phase of 2K-1C hypertensive rats whose R-A system was enhanced, captopril treatment further enhanced plasma renin activity and plasma angiotensin I and suppressed plasma angiotensin II while reducing blood pressure. Synthesis of the vascular proteins was almost identical. In the chronic phase of 2K-1C hypertensive rats whose R-A system was within normal limits, increased incorporation rates of 3H-proline into noncollagenous protein or collagen of mesenteric arteries were decreased by splanchnicotomy or beta-aminopropionitrile and hypertension was lowered. Captopril failed to reduce protein synthesis. CONCLUSIONS: An enhanced R-A system participates in the pathogenesis of the acute phase of 2K-1C hypertension while increased noncollagenous protein and collagen syntheses of small arteries appear to play some role in the etiology of the chronic phase of hypertension. PMID- 8709343 TI - Use of an anti-ras ribozyme to alter the malignant phenotype of a human bladder cancer cell line. AB - PURPOSE: In this study, we evaluated the ability of a ribozyme (catalytic RNA), which site specifically cleaves the mRNA of the activated H-ras gene, to alter the malignant phenotype of an invasive human bladder cancer cell line. MATERIALS AND METHODS: The human bladder cancer cell line EJ which contains the activated H ras gene was used in these studies. Cell lines with and without the anti-ras ribozyme were examined for their malignant potential in athymic (nude) mice by using an orthotopic model of bladder cancer. Endpoints evaluated included tumor take and animal survival. RESULTS: EJ tumors containing the anti-ras ribozyme showed a reduction in tumor take (35% versus 45%) and prolonged survival (74 days versus 65 days) compared with standard EJ cells. This survival advantage was not as pronounced as anticipated. To evaluate this finding, we examined the tumor from mice originally inoculated with the ribozyme-containing cell line to determine if the ribozyme was still present. Approximately 60% of the animals had lost ribozyme expression. Animals that maintained ribozyme expression had a mean survival of 81 +/- 4 days which was significantly prolonged compared with control mice (65 +/- 5 days). CONCLUSION: This study suggests that the invasive phenotype is blunted with the anti-ras ribozyme, delaying but not abolishing the metastatic phenotype. These results further delineate the roles of ras genes in malignancy and demonstrate that ribozymes may be a powerful tool for exploring the role of individual oncogenes and may be used as anticancer agents. PMID- 8709344 TI - Mycobacterium cell wall: an alternative to intravesical bacillus Calmette Guerin (BCG) therapy in orthotopic murine bladder cancer. AB - PURPOSE: The antitumor effect of intravesical mycobacterium cell wall (MCW) therapy on orthotopic and heterotopic bladder tumors in the mouse was assessed with magnetic resonance imaging (MRI). MATERIALS AND METHODS: The live bacillus Calmette Guerin (BCG) organism was replaced with a cell wall extract derived from the outer capsule of Mycobacterium phlei. This alternative form of intravesical therapy was used with the aim of reducing the toxicity associated with the live mycobacterium organism without compromising efficacy. Response to multiple doses of intravesical MCW and BCG was assessed in mice with established MBT-2 tumors after transurethral tumor implantation. RESULTS: Serial MRI of BCG-treated mice revealed significant tumor regression. The MR images correlated well with the corresponding histology of the whole mount bladder sections. Treatment with MCW also resulted in significant inhibition of tumor growth compared with control untreated animals (p < 0.05) although the antitumor effect was less pronounced than that of live BCG. Treatment was well tolerated in the MCW group with no apparent ill effects. Flow cytometric (FCM) analysis of bladder washings with phenotype-specific monoclonal antibodies revealed predominantly a CD3+ T cell infiltrate in the control and BCG-treated as well as the MCW-treated mice. The CD4+ (helper/inducer) subset of T cells predominated over the CD8+ (suppressor/cytotoxic) subset in both the BCG- and MCW-treated animals, and the CD4+/CD8+ ratio in both of the treated groups differed significantly from that of the control untreated groups. CONCLUSION: Intravesical MCW appears to invoke a similar inflammatory response in the mouse bladder mucosa as the live BCG organism and retains an antitumor action. It deserves further evaluation as a potential antitumor agent against bladder cancer. A Phase II clinical trial is now underway. PMID- 8709345 TI - Elevated estrogen receptor expression in human prostatic stromal cells by androgen ablation therapy. AB - PURPOSE: The presence of estrogen receptors (ER) in human prostatic tissue is a longstanding, controversial issue. In a few experimental animal models androgen deprivation was shown to be associated with a spontaneous increased ER expression in prostatic tissue. We intended to study whether these observations also apply to human prostatic tissue. MATERIALS AND METHODS: Estrogen receptor expression by stromal and glandular cells was studied by immunohistochemistry in prostatectomy specimens of 21 patient with prostate cancer, treated for 3 months with a luteinizing hormone-releasing hormone (LHRH) agonist and flutamide. In addition 2 patients treated with estrogens were also examined. For comparison, ER expression was also studied in a series of 18 prostatectomy specimens of untreated patients. RESULTS: The specimens of patients treated with androgen blockade showed atrophic changes of the gland as well as basal cell hyperplasia, features characteristic for this therapy. Although stromal cells of prostatectomy specimens from untreated patients were largely ER negative, those of patients exposed to androgen ablation therapy or estrogen therapy had an intense nuclear ER expression in a great number of stromal cells around prostatic glands. Sporadic epithelial cells lining the glands displayed some nuclear ER expression. Prostatic glands from treated patients with basal cell hyperplasia lacked ER expression. In all treated and untreated cases the carcinoma cells were ER negative. CONCLUSIONS: Androgen deprivation leads to an upregulation of stromal ER expression in human prostate. Estrogen-induced morphological epithelial changes could be explained by a paracrine interaction between stromal and epithelial cells. PMID- 8709346 TI - Ureteral cell cultures. I. Characterization and cellular interactions. AB - PURPOSE: To further understand the biology of ureteral cells, we studied the growth characteristics and in vitro cellular interactions of human ureteral uroepithelial cells, smooth muscle cells and myofibroblasts. MATERIALS AND METHODS: The proliferation, morphology and immunohistochemical characteristics of human ureteral cells grown in vitro were evaluated under varying conditions. RESULTS: The growth and morphology of ureteral cells were dependent upon media characteristics, especially the calcium concentration and presence of epidermal growth factor and bovine pituitary extract. Cells demonstrated specific stimulatory interactions via both soluble and insoluble factors. Most important, uroepithelial cells and smooth muscle cells displayed reciprocal enhancement of growth. CONCLUSIONS: The favorable interactions between ureteral cell types in vitro have implications for future work involving these cells. PMID- 8709347 TI - Pax-2: a developmental gene constitutively expressed in the mouse epididymis and ductus deferens. AB - PURPOSE: To characterize expression of the transcriptional activator, Pax-2, in the mouse lower genitourinary tract. MATERIALS AND METHODS: Expression of Pax-2 was studied by Northern analysis, ribonuclease protection and immunohistochemistry. RESULTS: Immunostaining revealed localized expression in the epithelium of the ductus deferens and epididymis at all time points from birth to adulthood. Expression in these structures in adult mice was confirmed by Northern analysis and ribonuclease protection assays. CONCLUSION: Pax-2 is a transcriptional regulator expressed in the epithelium of the ductus deferens and epididymis and may be a regulator of epithelial genes involved in sperm maturation and support. PMID- 8709348 TI - Evaluation of mRNAs encoding muscarinic receptor subtypes in human detrusor muscle. AB - PURPOSE: The present study evaluated the muscarinic receptor subtypes corresponding to m1 to m5 genes in human detrusor muscle. MATERIALS AND METHODS: The mRNAs encoding m2 and m3 subtypes were assessed by reverse transcription (RT) polymerase chain reaction (PCR). The amounts of cDNA synthesized from m2 and m3 mRNAs were measured by using subcloned plasmid DNAs. The distribution of m2 and m3 mRNAs in detrusor was estimated by comparing the amount of m2 cDNA with that of m3 cDNA. RESULTS: The m2 mRNA:m3 mRNA ratio was 1.06:1.00 in human detrusor. In the cryostat sections of human detrusor, the presence of both m2 and m3 mRNAs was confirmed by in situ hybridization. However, the RT-PCR products derived from m1, m4 and m5 subtype mRNAs were not detected. CONCLUSION: These results suggest that human detrusor muscle coexpresses muscarinic m2 and m3 receptors and that the populations of the 2 subtypes are not significantly different. PMID- 8709349 TI - This month in investigative urology. Commentary on incisions, stents and healing. PMID- 8709350 TI - Familial transitional cell carcinoma. AB - PURPOSE: Bladder cancer is a common malignancy, and a frequent cause of urological consultation and surgical intervention. Except for smoking and certain occupational exposures, the etiology of bladder cancer is largely unknown. Although the majority of patients with bladder cancer do not have a family history of transitional cell carcinoma of the urinary tract, the study of familial transitional cell carcinoma may lead to knowledge of the pathogenesis of this disease. MATERIALS AND METHODS: To evaluate the current understanding of familial transitional cell carcinoma, we reviewed the contemporary literature for case reports and epidemiological studies about this disease. RESULTS: Numerous case reports document the clustering of transitional cell carcinoma in families, several of which demonstrate an extremely early age at onset of disease, which argues in favor of a genetic component to familial transitional cell carcinoma. The results of large epidemiological studies also suggest the existence of familial transitional cell carcinoma, and first degree relatives appear to have an increased risk for disease by a factor of 2. Familial clustering of smoking does not appear to be the cause of this increased risk. CONCLUSIONS: Familial transitional cell carcinoma may be the result of a genetically transmitted predisposition to disease, at least in some affected families. Further studies are required to identify candidate genes that may be responsible for this form of bladder cancer. PMID- 8709351 TI - Randomized, prospective, controlled study comparing radical prostatectomy alone and neoadjuvant androgen withdrawal in the treatment of localized prostate cancer. Canadian Urologic Oncology Group. AB - PURPOSE: A prospective, multicenter, randomized study was done to test the hypothesis that neoadjuvant androgen withdrawal decreases the incidence of positive margins following radical prostatectomy for localized prostate cancer. MATERIALS AND METHODS: Observations were made of 213 patients randomized to undergo radical prostatectomy alone (101) or to receive a 12-week course of 300 mg. cyproterone acetate daily followed by surgery (112). Groups were similar at baseline in terms of clinical stage, serum prostate specific antigen and Gleason score. Of 192 patients available for efficacy analysis 9 had stage T1b, 8 stage T1c, 63 stage T2a, 36 stage T2b and 76 stage T2c disease. RESULTS: One or more positive surgical margins were found in 59 of 91 patients (64.8%) in the surgery only group compared to 28 of 101 (27.7%) in the cyproterone acetate group (p = 0.001). Patients who received preoperative therapy had a statistically significantly lower rate of apical margin involvement than those who did not (17.8 versus 47.8%, respectively, p < 0.0001). There was no statistically significant difference in surgical (p = 0.8645) or postoperative (p = 0.173) complications between the 2 groups. CONCLUSIONS: Neoadjuvant androgen withdrawal with a 12-week course of 300 mg. cyproterone acetate daily results in a lower rate of positive margins without adversely affecting postoperative recovery. The impact on patient survival will be determined by long-term followup. PMID- 8709352 TI - Doppler ultrasonography in evaluation of potential live kidney donors: a prospective study. AB - PURPOSE: We compared the accuracy of Doppler ultrasonography and intra-arterial digital subtraction angiography for preoperative visualization of the number of renal arteries in potential live kidney donors. MATERIALS AND METHODS: Intra arterial digital subtraction angiography and Doppler ultrasonography were used to study the renal anatomy of 64 consecutive kidney donors. Doppler studies were performed without knowledge of the angiographic findings. All donors underwent nephrectomy, and the number of renal arteries at nephrectomy was compared with that predicted by intra-arterial digital subtraction angiography and Doppler ultrasonography. RESULTS: One renal artery was found in 53 harvested kidneys, 2 in 10 and 3 in only 1. Analysis of the data revealed sensitivity 96.2% and specificity 100% for intra-arterial digital subtraction angiography versus sensitivity 100% and specificity 60% for Doppler ultrasound. Overall accuracy was 96.8% for angiography versus 93.7% for Doppler ultrasound, compared to 96.2 and 100%, respectively, for single, and 100% and 60%, respectively, for double renal arteries (p < 0.03). Both tests were interpreted as showing double vessels in the case with triple renal arteries. CONCLUSIONS: Although Doppler ultrasonography is accurate in delineation of single renal arteries, its limited ability to identify multiple arteries should make one cautiously optimistic regarding its current use. PMID- 8709353 TI - Insertion of a double pigtail ureteral stent for the prevention of urological complications in renal transplantation: a prospective randomized study. AB - PURPOSE: Urologists successfully use ureteral stents to protect the ureterovesical anastomosis in nontransplant patients. MATERIALS AND METHODS: We determined the value of ureteral stents in transplant patients. The frequency of urological complications (leaks, obstructions and urinary tract infections) was compared in a prospective randomized series of 194 kidney transplantations (97 with and 97 without a double pigtail ureteral stent). RESULTS: In the stent group 1 patient had a urinary leak and 35 had urinary tract infections (including 2 cases of Corynebacterium cystitis). In the no stent group 6 patients had urinary leaks, 4 had obstructions and 32 had urinary tract infections. The 1-year patient and graft survival rates were similar in both groups, and renal function at 1 year was also similar (229 versus 208 mumol./l. creatinine in the stent and no stent groups, respectively). A small number of stent related complications occurred (2 stent breakages and 1 stent migration). No stones formed in any case. CONCLUSIONS: Ureteral stent insertion significantly decreases the rate of vesicoureteral leakage and obstruction in renal transplantation. PMID- 8709355 TI - Renal transplantation--improving the quality of life. PMID- 8709354 TI - Transplantation into the long-term defunctionalized bladder. AB - PURPOSE: We evaluated the outcome of transplantation into a long-term defunctionalized bladder. MATERIALS AND METHODS: Since 1985 we performed transplantation in 5 dialysis dependent patients after excision of the ileal conduit and native kidneys. The bladder was evaluated before transplantation with cystoscopy, voiding cystography, urodynamics and demonstration of continence. Bladder rehabilitation was accomplished by cycling through a suprapubic tube or urethral catheter, and no bladder augmentation was done. All patients received antibiotic prophylaxis for several years. RESULTS: Five male recipients underwent transplantation at ages 22, 29, 30, 31 and 55 years, and they had had a defunctionalized bladder for 15, 17, 23, 25 and 26 years. All patients were alive with a functioning allograft at 6, 84, 90, 112 and 120 months after transplantation. Current serum creatinine values are 1.2, 1.6, 1.8, 2.3 and 2.5 mg./dl. Median urodynamic values before and after transplantation were bladder capacity 60 and 300 cc, respectively, peak flow rate 5 and 18 cc per second, respectively, and post-void residual 20 and 15 cc, respectively. One patient required self-catheterization. CONCLUSIONS: Renal transplantation into a long term defunctionalized bladder can be performed safely in carefully selected patients. Bladder function and continence should be confirmed before transplantation using a program of progressive bladder rehabilitation. PMID- 8709356 TI - Long-term results with single pediatric donor kidney transplants in adult recipients. AB - PURPOSE: We investigated whether transplantation of single pediatric donor kidneys into adults leads to an increased incidence of functional allograft impairment and complications, as previously reported. MATERIALS AND METHODS: To evaluate long-term functional outcome using single pediatric donor kidneys 60 adults (study group) who underwent transplantation between March 1973 and December 1988 using single pediatric donor kidneys 6 years old or younger (mean donor age plus or minus standard deviation 41.1 +/- 17.9 months) were compared to 58 matched adults (control group) who underwent transplantation with adult kidneys (mean donor age 29.7 +/- 10.8 years). The groups were identical for era of transplantation, recipient age, sex and followup (82 versus 83 months). RESULTS: There was no difference in patient survival between the study and control groups (p = 0.26). In the study group there were an increased requirement for early dialysis (45 versus 24%, p = 0.02), a higher incidence of proteinuria (greater than 0.8 gm./24 hours, 67 versus 48%, p = 0.04) and a higher incidence of rejection within the first 6 months (80 versus 64%, p = 0.05). There was also an increased incidence of graft failure from acute rejection in the study group. Early differences in serum creatinine levels in the 2 groups dissipated after 3 months. Renal allograft histopathology revealed no significant difference in the incidence of focal segmental glomerulosclerosis in the study versus control groups after transplantation (22.9 versus 13.3%, p = 0.70). However, focal segmental glomerulosclerosis manifested sooner after transplantation in study than control patients (mean 37 versus 82 months). After transplantation proteinuria developed in study patients with focal segmental glomerulosclerosis at a mean of 4.6 months compared to 31.8 months in controls with post-transplant focal segmental glomerulosclerosis. Graft survival in the study group was superior when cyclosporine was given rather than conventional noncyclosporine based immunosuppression. Five-year graft survival rates were 48 versus 44% for cyclosporine treated and 33 versus 44% for conventionally treated study versus control patients. CONCLUSIONS: These data suggest that with cyclosporine immunosuppression transplanting single pediatric kidneys into adults yields the same long-term functional graft outcome as adult donor kidneys. PMID- 8709357 TI - The use of ungating with the Medstone lithotriptor. AB - PURPOSE: Ungating using the Medstone* lithotriptor by 200 urologists was evaluated. MATERIALS AND METHODS: During 1994, 3,288 patients were treated by 200 urologists at 46 sites in 6 upper midwest states using 5 fixed and 3 mobile Medstone lithotriptors. Ungating was used with 58 treatments in 57 asymptomatic patients (1.8%) due to irregular cardiac rhythm in 48 caused by a bundle branch block (11), atrial fibrillation (10), slow heart rate (6) and irregular cardiac complex (21); all 48 cases were clinically insignificant, and because of urologist choice in 9 with normal cardiac rhythm (10 treatments). The cardiac simulator used for ungated lithotripsy was set at 85 shocks per minute for irregular cardiac rhythm and at 120 shocks per minute for elective use. RESULTS: The 48 treatments in patients with clinically insignificant irregular cardiac rhythm (average age 66.4 years) were performed during an average of 41 minutes of shock time. One patient had clinically significant cardiac arrhythmia that resolved with gating. The 10 elective treatments were performed during an average of 34 minutes of shock time in patients an average of 60.4 years old. The 3,231 gated treatments were performed during an average of 40 minutes of shock time in patients an average of 51 years old. CONCLUSIONS: Ungating was safe and effective in allowing patients with an irregular cardiac rhythm to be treated with the same shock time as gated cases (normal cardiac rhythm). PMID- 8709358 TI - Ureteroscopy with intravenous sedation for treatment of distal ureteral calculi: a safe and effective alternative to shock wave lithotripsy. AB - PURPOSE: We evaluated the safety, efficacy and patient tolerance of ureteroscopy with intravenous sedation for treatment of distal ureteral calculi. MATERIALS AND METHODS: Ureteroscopy with intravenous sedation was performed in 68 patients, with small distal ureteral calculi requiring treatment and in 2 with middle or upper third ureteral calculi. Intravenous sedation and analgesia were achieved with a combination of fentanyl and midazolam. Balloon dilation of the ureter was performed in 38 patients. Ureteroscopy was performed with a 6F rigid ureteroscope. RESULTS: Stone removal was successful in 66 patients (97%). There were no ureteral perforations in patients undergoing successful stone removal and no complications related to intravenous sedation. Tolerance of the procedure was good in 81% of the patients (including 72% of 36 men and 94% of 32 women undergoing ureteroscopy for distal ureteral calculi) and fair in 19%. CONCLUSIONS: In select patients with small distal ureteral calculi ureteroscopy offers a safe, effective and well tolerated alternative to shock wave lithotripsy. PMID- 8709359 TI - Recurrence rate and risk factors for urinary calculi after extracorporeal shock wave lithotripsy. AB - PURPOSE: We determined the recurrence rate after extracorporeal shock wave lithotripsy (ESWL*) and evaluated its associated risk factors, such as patient sex and age, volume of daily water intake, related diseases, family history of urinary calculi, number, burden and location of stones, residual stones after ESWL, serum uric acid level before ESWL and so forth. MATERIALS AND METHODS: We conducted a retrospective study of 841 patients who underwent ESWL for primary treatment of urinary calculi between 1985 and 1987. Charts were reviewed for stone characteristics, outcomes and associated laboratory data. Followup 6 to 9 years later was done by mail and telephone interviews. RESULTS: Followup data were available for 436 patients (52%). Overall, stones recurred in 51% of the 436 patients at a mean followup of 7.1 years. Late recurrences were common, and were noted in as many as 70% of the patients after 9 years. This rate is greater than what has been previously reported after other treatments. Cox regression analysis indicated that the major risk factors associated with recurrence were male gender (relative risk 1.406, p < 0.05) and multiple stones (relative risk 1.777, p < 0.05). CONCLUSIONS: The frequent late recurrences support the speculation that ESWL might lead to greater recurrence rates. Therefore, the cost-effectiveness of ESWL versus other modalities for stone removal warrants more long-term evaluation. PMID- 8709360 TI - Dietary manipulation with lemonade to treat hypocitraturic calcium nephrolithiasis. AB - PURPOSE: Pharmacological treatment of hypocitraturic calcium nephrolithiasis requires as many as 12 tablets, or numerous crystal packages or liquid supplements taken throughout the day. In addition to added cost, this cumbersome regimen decreases patient compliance, which may increase stone recurrence rates. We evaluated the urinary biochemical effects of dietary citrate supplementation in hypocitraturic calcium stone formers in an attempt to decrease or eliminate the need for pharmacological therapy. MATERIALS AND METHODS: A total of 12 patients who were either noncompliant with or intolerant of pharmacological citrate therapy supplemented their routine diet with citrate in the form of lemonade, consisting of 4 ounces of reconstituted lemon juice (5.9 gm. citric acid) mixed with tap water to a total volume of 2 l. and consumed at uniform intervals throughout the day. Urine specimens (24-hour) were obtained for biochemical analysis after 6 days of lemonade therapy and compared to pre lemonade baseline values. RESULTS: Of the 12 patients 11 had increased urinary citrate levels during lemonade therapy (average 204 mg. per day). Average levels increased from 142 mg. daily (range less than 10 to 293) at baseline to 346 mg. daily (range 89 to 814) after treatment (p < 0.001). Daily total urinary volumes were similar (2.7 versus 2.9 l.). Seven of 12 patients became normocitraturic while consuming lemonade. Urinary calcium excretion decreased an average of 39 mg. daily, while oxalate excretion was unchanged. The lemonade mixture was well tolerated. Two patients complained of mild indigestion that did not require cessation of therapy. CONCLUSIONS: Citrate supplementation with lemonade increased urinary citrate levels more than 2-fold without changing total urinary volume. Lemon juice, which contains nearly 5 times the concentration of citric acid compared to orange juice, is an inexpensive and well tolerated dietary source of citrate. Lemonade therapy may improve patient compliance, and may be useful as adjunctive treatment for patients with hypocitraturic calcium nephrolithiasis. PMID- 8709361 TI - Nephrolithiasis--solutions for emerging problems. PMID- 8709362 TI - Intracorporeal lithotripsy with the holmium:YAG laser. AB - PURPOSE: Preliminary evaluations of the holmium:YAG laser have demonstrated a variety of potential urological applications, including ablation of soft tissue lesions as well as stone fragmentation. We present our experience with the holmium:YAG laser for intracorporeal lithotripsy of urinary calculi. MATERIALS AND METHODS: During a 24-month period 75 patients underwent 79 laser procedures, including retrograde ureteroscopy for ureteral calculi (71) and fragmentation of caliceal stones remote from the nephrostomy tract during percutaneous nephrolithotripsy (8). RESULTS: Complete stone fragmentation without need for additional procedures or lithotripsy was achieved in 85% of the cases. Treatment failures included 1 case of stone migration, 7 incomplete fragmentation requiring other lithotripsy devices and 3 laser malfunction. One ureteral perforation occurred when the laser was activated without direct visual guidance. CONCLUSIONS: The holmium:YAG laser has demonstrated its efficacy as a method of intracorporeal lithotripsy. Advantages include ability to fragment stones of all composition, and the multipurpose, multispecialty applications of the holmium wavelength. This laser has potential soft tissue effects, and careful attention to technique during lithotripsy is required to avoid ureteral wall injury. PMID- 8709363 TI - Detubularized isolated ureterosigmoidostomy: description of a new technique and preliminary results. AB - PURPOSE: A technique is described to improve the evacuation pattern and, accordingly, the life-style of patients with ureterosigmoidostomy. MATERIALS AND METHODS: An inverted U-shaped sigmoid colon is detubularized, and the left colon is fixed in continuity to the posterior wall of the rectal ampulla in line with the anorectal canal. The ureters are reimplanted into the sigmoid pouch using the nipple technique. RESULTS: All 15 patients followed for 3 to 18 months postoperatively passed clear urine and solid feces separately, with good anal control and at convenient periods (urine 3 to 6 times and solid feces once daily). Urographic studies showed stabilization of renal morphology in 26 units, improved function in 3 and deterioration in 1. Dynamic study of the rectum in 3 patients showed a capacious rectosigmoid reservoir with low pressure. CONCLUSIONS: The technique of detubularized isolated ureterosigmoidostomy has important advantages over conventional ureterosigmoidostomy. Patients pass urine and feces separately and at convenient intervals with good anal control. The upper urinary tract is well preserved, and there is potentially less risk of colonic carcinogenesis. PMID- 8709364 TI - The Kock ileal neobladder: updated experience in 295 male patients. AB - PURPOSE: Since 1986 orthotopic lower urinary tract reconstruction using the Kock ileal neobladder has been our diversion of choice in patients undergoing cystectomy. We report on the first 295 male patients undergoing this procedure from May 1986 through December 1993. MATERIALS AND METHODS: Complications were assessed, tabulated, subdivided into early (3 months or less postoperatively) and late types, and further categorized with respect to relationship to neobladder construction. Continence was individually evaluated via a detailed patient questionnaire. RESULTS: The pouch related early and late complication rates were 7.2 and 11.6%, respectively, and pouch related abdominal reoperation rates were 0.0 and 1.4%, respectively. Analysis of late pouch related complications revealed 4.1% stone formation and 2.4% afferent nipple stenosis rates, and only 1 case (0.3%) of ileal urethral anastomotic stricture. Of the patients 87 and 86% reported good or satisfactory daytime and nighttime continence, respectively. With regard to age, while overall continence was similar, a significantly greater percentage of patients younger than 70 years experience good daytime and nighttime continence relative to the older counterparts. Of the patients 5% perform regular intermittent catheterization and 2.7% required an artificial urinary sphincter due to unacceptable continence. CONCLUSIONS: The Kock orthotopic ileal neobladder can be constructed with minimal morbidity and extraordinary functional results. We strongly advocate its use when possible. PMID- 8709365 TI - The Le Bag orthotopic urinary diversion. AB - PURPOSE: We reviewed our experience with Le Bag orthotopic urinary diversion in 38 cases. MATERIALS AND METHODS: Between April 1990 and January 1995, 38 men underwent radical cystectomy and Le Bag urinary diversion for treatment of bladder cancer. Approximately 20 cm. each of detubularized ileum and cecum were used to construct the pouch. A total of 22 pouches was fashioned with absorbable staples. In 21 cases freely refluxing Bricker ureterointestinal anastomoses were used. RESULTS: There was no significant difference in major complication rates in the hand sewn versus stapled anastomosis groups, and there were no complications specifically related to the use of staples. There were 3 episodes of febrile urinary tract infection: 2 in the Bricker group and 1 in the tunneled anastomosis group. There was no significant difference between the 2 groups with respect to ureteral obstruction. The daytime continence rate was 91%, and 80% of the patients are completely dry or have only mild incontinence at night. Most patients had mild hyperchloremic metabolic acidosis postoperatively as evidenced by a decrease in median serum bicarbonate level (28 versus 24 mmol./l.). This difference appears to be related to pouch length (r = 0.58, p = 0.0002). CONCLUSIONS: We conclude that the Le Bag technique is a technically feasible form of urinary diversion with functional results similar to other forms of orthotopic diversion. Use of absorbable staples simplifies pouch construction without increasing complications. This form of urinary diversion is associated with hyperchloremic metabolic acidosis, which is related to pouch length. PMID- 8709367 TI - Continent urinary reconstruction--method-to-patient matching. PMID- 8709366 TI - Can gastric pouch as orthotopic bladder replacement be used in adults? AB - PURPOSE: We studied orthotopic bladder replacement with stomach in adults with bladder carcinoma, with emphasis on possible subsequent complications, such as incontinence, the dysuria or hematuria syndrome, ulceration or perforation. We attempted to prove a correlation between stabilized urine pH and hypergastrinemia. MATERIALS AND METHODS: Since June 1992, 19 men between 53 and 73 years old underwent surgery for nonmetastatic grade 3 bladder carcinoma according to our new technique. RESULTS: Patient outcome was good. Possible complications are discussed, such as incontinence, hypergastrinemia and ulcerations. CONCLUSIONS: Sufficient pouch capacity is achieved with our proposed operative technique. Urine pH must be monitored to prevent possible ulcerations and perforations. PMID- 8709368 TI - Urinary epitectin (MUC-1 glycoprotein) in the menstrual cycle and interstitial cystitis. AB - PURPOSE: We compared interstitial cystitis and control urine specimens for epitectin (MUC-1 glycoprotein), an epithelial mucin. MATERIALS AND METHODS: Urinary epitectin was measured in 28 patients with interstitial cystitis and 26 healthy controls. Ten controls provided multiple urine samples to determine whether urinary epitectin changes with the menstrual cycle. RESULTS: Epitectin levels were stable throughout the menstrual cycle. Interstitial cystitis cases had decreased urinary epitectin-to-creatinine ratios (mean 3.89 versus 6.38 micrograms./mg. creatinine for controls, p = 0.0035) and epitectin concentrations (mean 1.96 versus 4.30 micrograms./ml., respectively, p = 0.0005). CONCLUSIONS: Decreased mean urinary epitectin levels may reflect a cause (epithelial mucin deficiency) or a consequence of interstitial cystitis. PMID- 8709369 TI - Increased urinary levels of Tamm-Horsfall glycoprotein suggest a systemic etiology of interstitial cystitis. AB - PURPOSE: We investigated the role of Tamm-Horsfall protein in interstitial cystitis. MATERIALS AND METHODS: Urinary Tamm-Horsfall protein excretion was analyzed in interstitial cystitis patients and controls, and bladder biopsy specimens were stained immunohistochemically for Tamm-Horsfall protein. RESULTS: Urinary Tamm-Horsfall protein levels in 28 women with interstitial cystitis were statistically significantly greater than those in 25 female controls. No positive staining for Tamm-Horsfall protein was demonstrated in the bladder tissue from 10 interstitial cystitis cases. CONCLUSIONS: The results support the notion that interstitial cystitis may have a systemic etiology. In addition, this assay might have clinical value in the diagnosis of interstitial cystitis. PMID- 8709370 TI - Intravesical capsaicin for treatment of severe bladder pain: a randomized placebo controlled study. AB - PURPOSE: Present therapeutic approaches to control bladder pain are clinically and scientifically unsatisfactory, and pain in the lower urinary tract remains a challenge even to the skilled urologist. A randomized placebo controlled study was done to evaluate intravesical capsaicin for severe bladder pain. Followup was 6 months. MATERIALS AND METHODS: A total of 36 patients was prospectively randomized into those receiving 10 microM. intravesical capsaicin twice weekly for 1 month (group 1) or placebo (group 2). All patients had pelvic pain for at least 6 months, and had no urinary tract infection within the last 3 months, functional disorders of the lower urinary tract, or other vesical or urethral pathology. Pretreatment voiding pattern and pain score were recorded. Patients were evaluated immediately at the end of treatment (primary end point) and 6 months later (secondary end point). RESULTS: Both groups were adequately homogeneous with regard to age, sex ratio, duration of disease, voiding pattern and pain score. At both end points group 1 had significant improvement in frequency and nocturia but no improvement in urgency. No change was noted in group 2. A significant decrease in pain score was found in group 1 at the primary (mean plus or minus standard deviation 3.22 +/- 0.42, p < 0.01) and secondary (3.83 +/- 0.47, p < 0.01) end points compared to before treatment (5.61 +/- 0.40, chi-square with 2 degrees of freedom 29.25, p < 0.0001). A significant improvement was also observed in the placebo group, in which the pretreatment pain score (5.47 +/- 0.37) was decreased at the primary (4.47 +/- 0.36, p < 0.01) and secondary (4.48 +/- 0.34, p < 0.01, chi-square with 2 degrees of freedom 12.71, p < 0.002) end points. There were no statistically significant differences between the 2 groups. CONCLUSIONS: We confirmed the beneficial effect of intravesical instillation of capsaicin on voiding pattern in patients with hypersensitive disorders (frequency and nocturia). We could not confirm improvement in pain score after capsaicin treatment compared to placebo. Possibly a larger dose of capsaicin would be more effective in controlling pain and neurological disease of the bladder. PMID- 8709371 TI - Transforming growth factors-beta 1 and beta 2 in serum and urine from patients with bladder carcinoma. AB - PURPOSE: Transforming growth factors-beta (TGF-beta) are cellular regulators and potent angiogenic factors. We determined serum and urinary levels of TGF-beta 1 and beta 2 in patients with bladder carcinoma to study a correlation with tumor stage, grade and metastatic spread. MATERIALS AND METHODS: Using commercial immunoassays TGF-beta 1 and beta 2 were determined in serum and urine samples from 57 bladder cancer patients and 18 healthy controls. RESULTS: Serum TGF-beta 1 levels were significantly elevated in 21 patients with invasive bladder cancer (61.5 ng./ml.) compared to 18 healthy controls (36.3 ng./ml.), whereas serum TGF beta 1 levels in 36 patients with superficial bladder tumors were within the normal range (33.4 ng./ml.). Serum TGF-beta 1 was increased in 27 patients with grade 3 tumors (55.7 ng./ml.), compared to 16 with grade 1 and 14 with grade 2 tumors (32.6 and 33.3 ng./ml., respectively). By contrast, serum TGF-beta 2 levels were not different from those of controls. No significant increase in serum TGF-beta 1 and beta 2 could be found in patients with lymph node metastases. In urine specimens there was no significant correlation of TGF-beta 1 and beta 2 with tumor stage. CONCLUSIONS: Our results suggest that elevated serum TGF-beta 1 may be relevant for diagnosis of bladder cancer and further studies are warranted. PMID- 8709372 TI - Deoxyribonucleic acid measurements in transitional cell carcinomas: comparison of flow and image cytometry techniques. AB - PURPOSE: We compared the results of deoxyribonucleic acid (DNA) ploidy determinations performed by flow cytometry and image cytometry in transitional cell carcinomas of the bladder. MATERIALS AND METHODS: In 81 cases of transitional cell carcinoma of the bladder DNA indexes were measured by flow cytometry of bladder washings and tumor tissue samples, and by image cytometry of imprints from tumor tissue samples. RESULTS: There was good correlation between bladder washings and tissue samples analyzed by flow cytometry in 68 cases but aneuploidy was missed with bladder washings in 13 (16%). There was also good correlation between flow and image cytometry in 75 cases (92.59%) regarding the detection of aneuploidy. There was agreement between detection of aneuploidy and DNA index in 45 cases (55.5%), while both methods detected an aneuploid population in 30 (37%) but there was disagreement regarding DNA index. Aneuploid populations were missed by flow cytometry in 6 cases (7.4%). Furthermore, in 10 cases peridiploid peaks were found on the image cytometry histograms, which were not visible on flow cytometry. However, it was not possible to assess accurately if these were true peridiploid populations. CONCLUSIONS: There is good overall correlation between DNA content measured by flow and image cytometry but image cytometry has the advantage of visual discrimination, permitting preferential selection and analysis of tumor cells. However, certain problems remain with image cytometry, particularly in the case of peridiploid peaks, which cannot be classified accurately as showing true peridiploid or right shifted diploid populations. PMID- 8709373 TI - Transitional cell carcinoma--tantalizing issues of tumor biology. PMID- 8709374 TI - A randomized multicenter trial of adjuvant therapy in superficial bladder cancer: transurethral resection only versus transurethral resection plus mitomycin C versus transurethral resection plus bacillus Calmette-Guerin. Participating Clinics. AB - PURPOSE: A randomized multicenter trial was done to compare transurethral resection only to transurethral resection plus adjuvant mitomycin C and bacillus Calmette Guerin (BCG) instillation for treatment of superficial bladder cancer (stage pTa/1 grades 1 to 3 except primary stage pTa grade 1). MATERIALS AND METHODS: Included in the study were 337 patients with superficial stage pTa/1 grades 1 to 3 bladder cancer except primary stage pTa grade 1 tumors. One group underwent transurethral resection alone. Mitomycin C (20 mg./50 ml. sodium chloride) was given every 2 weeks during year 1 and once a month during year 2. BCG (120 mg/50 ml. sodium chloride was instilled once a week for 6 weeks and once a month for 4 months. RESULTS: At a median followup of 20.2 months, a decrease in recurrence rate was noted for both drug instillations compared to transurethral resection only. The relative risk of recurrence was 0.508 after mitomycin C and 0.618 after BCG instillation compared to transurethral resection alone. There was no significant difference between the mitomycin C and BCG instillations. The progression rate was comparable in all 3 therapy groups, with an estimated common progression rate of 4.22% per year. Side effects occurred most frequently during or after BCG instillation, most often consisting of cystitis. One patient required cystectomy because of ulcerating cystitis and a prostatic abscess subsequent to unsuccessful tuberculostatic therapy. There were no systemic complications. CONCLUSIONS: Our study showed a positive effect of adjuvant chemotherapy and immunotherapy on decreasing tumor recurrence rate. No influence was observed concerning progression rate, which was low overall. PMID- 8709375 TI - Clinicopathological evaluation of repeated courses of intravesical bacillus Calmette-Guerin instillation for preventing recurrence of initially resistant superficial bladder cancer. AB - PURPOSE: The efficacy of repeated courses of intravesical bacillus Calmette Guerin (BCG) for superficial bladder cancer was assessed with particular attention to initially resistant cases. MATERIALS AND METHODS: A total of 75 patients with stages Ta to T1b superficial transitional cell bladder carcinoma received 6 weekly instillations of 80 mg. Tokyo strain BCG in 40 ml. saline followed by 6 instillations at monthly intervals. If tumors recurred, another course of treatment was given with surgery. RESULTS: Of 17 patients (22.7%) with recurrent tumor at followup periods of up to 84 months 12 received an additional course of BCG instillations according to the same protocol after transurethral resection of bladder tumors, and 10 (83.3%) showed no further recurrence with or without additional surgery and BCG therapy after a median followup of 42.9 months. Thus, the overall success rate with this approach was 90.7% (68 of 75 patients). Comparison of patients with and without recurrence revealed a significant difference in number of tumors before therapy (p < 0.05), and a pronounced tendency (p = 0.00507) for recurrence after prior systemic chemotherapy or intravesical instillation. CONCLUSIONS: The results suggest that up to 3 courses of repeated intravesical instillation of BCG are effective even for cases that initially did not respond, and that best results may be achieved if no other prior chemotherapy has been attempted. PMID- 8709376 TI - A new staging system for transitional cell carcinoma involvement of the prostate and reconfirmation studies of intravesical therapy for superficial bladder cancer. PMID- 8709377 TI - One-stage anterior urethroplasty. AB - PURPOSE: A deficient urethral segment was replaced with penile skin during a 1 stage procedure in patients with a long, tight urethral stricture, multiple attempts at hypospadias repair or severe hypospadias and circumcision. MATERIALS AND METHODS: In 29 patients a pedicled circumferential strip of distal penile skin was used to construct a neourethral floor. The roof was formed by regeneration of the epithelium from the edges of the floor over Buck's fascia. In our series the urethra was reconstructed because of an anterior urethral stricture in 11 patients, multiple failed hypospadias repairs in 6 and severe hypospadias with circumcision in 12. RESULTS: A neourethra of sufficient caliber and length was constructed with minimal postoperative complications in all patients. There were 2 cases of urethrocutaneous fistula at the subcoronal region, 1 meatal stenosis, 1 persistent chordee and 1 small distal penile skin patch slough that required only prolonged dressings. Mean followup was 19 months. CONCLUSIONS: Our urethroplasty technique can be used to correct various types of anterior urethral stricture or hypospadias associated with insufficient penile or preputial skin. PMID- 8709378 TI - Penile sensitivity in patients with primary premature ejaculation. AB - PURPOSE: We investigated penile sensory levels in patients with primary premature ejaculation to determine whether there is an etiological basis for this condition. MATERIALS AND METHODS: Penile biothesiometry was performed in 120 patients with primary premature ejaculation without erectile dysfunction and neurological deficit, and in 66 normal potent male volunteers. Vibratory thresholds were recorded at the glans penis, penile shaft, scrotum and index finger using a biothesiometer. RESULTS: On the glans penis and penile shaft the values in patients with premature ejaculation were significantly less than those in normal potent men (p < 0.001). In normal potent men an age dependency of biothesiometric parameters was noted, with a significant increase in vibratory threshold at the glans penis and penile shaft in older patients. However, in patients with premature ejaculation there were consistently decreased values without age dependency at the glans penis and penile shaft (p > 0.05). CONCLUSIONS: Patients with primary premature ejaculation have penile hypersensitivity, which provides further implications for an organic basis of premature ejaculation. PMID- 8709379 TI - Long-term results of arterial bypass surgery for impotence secondary to segmental vascular disease. AB - PURPOSE: We evaluated the long-term results of arterial bypass surgery in impotent men carefully selected for nonatherosclerotic arterial vascular disease. MATERIALS AND METHODS: During a 7-year period only 11 of 1,352 impotent men (0.8%) were selected as ideal candidates for arterial vascular surgery. Initial evaluation included history, physical examination and a screening pharmacological erection test. Duplex ultrasonography was performed in patients who failed to respond adequately to pharmacological stimulation and were believed to be potential candidates for vascular surgery. Further evaluation included formal nocturnal sleep laboratory testing, dynamic infusion cavernosometry and arteriography. Dorsal artery bypass was performed in 9 patients and arterialization of the deep dorsal vein was done in 2. RESULTS: Average followup was 50 months (range 12 to 84). Initial followup duplex ultrasonography revealed a significant increase in cavernous artery peak systolic blood flow velocity (p < 0.001) and patent arterial anastomoses in all but 1 patient. The initial success rate without supplemental injection therapy was 82% and the final success rate was 64%. Even with long-term followup, 91% of the patients were improved from baseline, and were sexually active with (27%) or without (64%) injection therapy. CONCLUSIONS: Arterial bypass surgery can be successful in select impotent patients without generalized atherosclerosis or other risk factors for impotence. Further refinement of screening parameters may improve patient selection and long term results of vascular surgery. PMID- 8709380 TI - The hemodynamic influence of the ischiocavernosus muscles on erectile function. AB - PURPOSE: Special attention has been focused on the contribution of the ischiocavernosus muscles to intracorporeal pressure during erection. MATERIALS AND METHODS: Length of the ischiocavernosus muscles and corpora cavernosa was studied in 30 cadavers. RESULTS: A biomechanical model for calculating interference of the ischiocavernosus muscles in rigidity during the initial phase of penile erection is proposed. CONCLUSIONS: We found that, among other things, intracorporeal pressure can be influenced by the ischiocavernosus muscles during erection. PMID- 8709381 TI - A randomized prospective comparison of antibiotic tissue levels in the corpora cavernosa of patients undergoing penile prosthesis implantation using gentamicin plus cefazolin versus an oral fluoroquinolone for prophylaxis. AB - PURPOSE: We performed a prospective randomized trial comparing the efficacy, safety and cost of parenteral antibiotics and oral fluoroquinolones for prophylaxis in penile prosthesis surgery. MATERIALS AND METHODS: We prospectively randomized 20 consecutive patients undergoing penile prosthesis surgery to receive ofloxacin orally or gentamicin and cefazolin parenterally followed by cephradine orally. Intraoperatively corpora cavernosa tissue and simultaneous peripheral serum samples were evaluated for antibiotic levels. Median followup was 16 months (range 8 to 21, mean 15.35). RESULTS: There were no implant losses or reoperations and complications were comparable in the 2 groups. The difference in mean serum-to-tissue ratios of ofloxacin versus the combination of cefazolin and gentamicin was statistically significant (p < 0.03). The minimum inhibitory concentrations of ofloxacin met or exceeded those of the 2 most common offending organisms, Staphylococcus epidermidis and Escherichia coli, in 80% of patients, which was comparable to the results of the parenteral regimen. Cost savings of the medications alone were more than $250,000 in the ofloxacin group. By eliminating a hospital stay of the 25,000 cases of penile prosthesis placement in the United States yearly a total cost savings of more than $36 million would be realized. CONCLUSIONS: When oral ofloxacin is given for prophylaxis in penile implant surgery, the procedure may be performed on an outpatient basis and health care dollars are saved. PMID- 8709382 TI - Penile length in the flaccid and erect states: guidelines for penile augmentation. AB - PURPOSE: We provide guidelines of penile length and circumference to assist in counseling patients considering penile augmentation. MATERIALS AND METHODS: We prospectively measured flaccid and erect penile dimensions in 80 physically normal men before and after pharmacological erection. RESULTS: Mean flaccid length was 8.8 cm., stretched length 12.4 cm. and erect length 12.9 cm. Neither patient age nor size of the flaccid penis accurately predicted erectile length. Stretched length most closely correlated with erect length. CONCLUSIONS: Only men with a flaccid length of less than 4 cm., or a stretched or erect length of less than 7.5 cm. should be considered candidates for penile lengthening. PMID- 8709383 TI - Potpourri. PMID- 8709384 TI - Alkalization does not alleviate penile pain induced by intracavernous injection of prostaglandin E1. AB - PURPOSE: A prospective, randomized, double-blind study was done to determine whether alkalization of prostaglandin E1 decreases pain associated with intracavernous injection. MATERIALS AND METHODS: Ten subjects with a history of pain following at least 70% of injections of prostaglandin E1 received 2 injections of prostaglandin E1, 1 with and 1 without sodium bicarbonate. Patients reported the duration and intensity of pain following each injection. RESULTS: Comparing injections of prostaglandin E1 with and without bicarbonate, there were no significant differences in number of subjects reporting pain, or duration and intensity of pain. CONCLUSIONS: Addition of sodium bicarbonate does not alleviate penile pain associated with injection of prostaglandin E1. PMID- 8709386 TI - Project HOPE helps China build a new foundation for pediatric medicine. PMID- 8709385 TI - A piece of my mind. When you come into my room. PMID- 8709387 TI - HOPE springs eternal for medical and health volunteers in 29 countries on 5 continents. PMID- 8709388 TI - Improved screening for prostate cancer offers challenging new data to cope with high incidence. PMID- 8709389 TI - Acellular pertussis vaccine is licensed for infants. PMID- 8709391 TI - Surgeon General's report on physical activity and health. From the Centers for Disease Control and Prevention. PMID- 8709390 TI - Identification of HIV-1 group O infection--1996. From the Centers for Disease Control and Prevention. PMID- 8709392 TI - Meat intake and non-Hodgkin lymphoma. PMID- 8709393 TI - Meat intake and non-Hodgkin lymphoma. PMID- 8709394 TI - Cost savings and prescribing protocols for nonsteroidal anti-inflammatory drugs. PMID- 8709395 TI - Cost savings and prescribing protocols for nonsteroidal anti-inflammatory drugs. PMID- 8709396 TI - Regression of left ventricular hypertrophy. PMID- 8709397 TI - Regression of left ventricular hypertrophy. PMID- 8709398 TI - Perinatal HIV infection. PMID- 8709399 TI - Intensity of testing and invasive procedures. PMID- 8709400 TI - Acute renal failure and mortality. PMID- 8709401 TI - Short stature and growth hormone therapy. A national study of physician recommendation patterns. AB - OBJECTIVE: To determine current expert opinion and recommendations regarding the controversial issue of the use of growth hormone (GH) to treat short children who do not have classical GH deficiency (non-GHD children). STUDY DESIGN: Analysis of a national survey mailed to 534 US physician experts on the management of short stature (pediatric endocrinologists) with a response rate of 81.3%. MAIN OUTCOME MEASURE: The experts' GH treatment recommendations. RESULTS: The physicians reported that approximately 58% of their current patients undergoing GH therapy have classical GH deficiency, while 42% have other conditions. The proportion of physicians who recommended GH treatment of short non-GHD children ranged from 1% to 74% over all case scenarios presented. The likelihood of GH being recommended depended on the physiological growth characteristics of the child (ie, the child's height, growth rate, and predicted adult height), contingency factors (ie, strong family wishes or a reduction in GH cost), and physician beliefs (ie, the impact of short stature on well-being, the effectiveness of GH therapy). Each of these factors exerted highly significant, independent, and additive effects on decisions to recommend GH. CONCLUSION: Our results indicate that many pediatric endocrinologists consider GH treatment appropriate for selected short non-GHD children, going beyond current Food and Drug Administration-approved indications for GH. Decisions to recommend GH for a non-GHD child rest on a combination of medical, social, and perceptual factors; variations in treatment patterns stem from variations in these influences. Future GH use will likely be determined not only by the results of controlled trials, but also by family preferences, producer pricing, and physician perceptions of the value of height and GH therapy. PMID- 8709402 TI - Effects of pegorgotein on neurologic outcome of patients with severe head injury. A multicenter, randomized controlled trial. AB - OBJECTIVE: To evaluate outcome of patients with severe closed head injury treated with pegorgotein, a scavenger of oxygen-derived free radicals. DESIGN: Randomized, parallel, placebo-controlled, third-party-blind, multicenter trial, with a blinded, multicenter follow-up protocol. SETTING: Twenty-nine centers in the United States. PATIENTS: A total of 463 patients with severe closed head injury and a Glasgow Coma Scale score of 8 or less after resuscitation and stabilization. INTERVENTIONS: Patients received a single intravenous dose of placebo, 10 000 U/kg of pegorgotein, or 20 000 U/kg of pegorgotein within 8 hours after injury. OUTCOME MEASURES: The primary endpoint was the Glasgow Outcome Scale (GOS) score at 3 months after brain injury with GOS data trichotomized into good, fair, or poor outcome. Secondary efficacy endpoints included the Disability Rating Scale (DRS) and mortality. A secondary analysis was performed using GOS scores dichotomized into favorable and unfavorable outcomes. In a follow-up protocol at 6 months, GOS and DRS scores were again determined. RESULTS: Of 463 patients randomized, 162 received placebo; 149, pegorgotein 10 000 U/kg; and 152, pegorgotein 20 000 U/kg. Treatment groups were comparable with respect to demographic characteristics, mechanism of injury, and time to treatment. Pegorgotein was well tolerated at both dose levels. At month 3, the trichotomized analysis found no significant statistical difference in neurologic outcome between the pegorgotein and the placebo groups. Although differences were not statistically significant, there were more favorable outcomes and no increase in the number of deaths or vegetative states among the patients given pegorgotein, more subjects had good or favorable outcomes with the 10 000-U/kg dose than with the 20 000-U/kg dose or placebo, and less disability was observed with the 10 000 U/kg dose than with either the 20 000-U/kg dose or placebo. No differences in mortality rate or cause of death were found between the 10 000-U/kg and placebo groups at either month 3 or month 6. The only statistically significant difference between the groups was a decreased incidence of adult respiratory distress syndrome in the 10 000-U/kg group as compared with the placebo group (P<.015). CONCLUSIONS: In this clinical trial of 463 patients with severe head injury, no statistically significant difference in neurologic outcome or mortality was observed between patients treated with pegorgotein and those receiving placebo. PMID- 8709404 TI - Elevated parathyroid hormone-related peptide associated with lactation and bone density loss. AB - OBJECTIVE: To investigate the hypothesis that parathyroid hormone-related peptide (PRHrP) may be involved with bone loss and recovery as a means of providing adequate calcium and phosphate to infants. DESIGN: An 18-month prospective cohort study. SETTING: General community setting with recruitment occurring at birthing education classes. PARTICIPANTS: Volunteer sample of 115 postpartum healthy women aged 20 to 40 years, and 0 or 1 parity prior to parturition with no intent to breast-feed or intent to breast-feed at least 6 months. MAIN OUTCOME MEASURES: Parathyroid hormone-related peptide, prolactin, estradiol, 1,25-dihydroxyvitamin D, 24-hydroxyvitamin D, femoral bone mineral density, and bone turnover markers were measured in 115 postpartum women at 2 weeks, 2 months, 4 months, 6 months, 12 months, and 18 months postpartum. Lumbar bone mineral density was measured at 2 weeks, 6 months, 12 months, and 18 months postpartum. RESULTS: Elevated PTHrP values were significantly associated (P<.001) with breast-feeding status, elevated prolactin levels, and lower serum estradiol levels, conditions occurring during lactation. Furthermore, elevated PTHrP levels were negatively and significantly associated (P<.01) over time with bone mineral density change at both the spine and the femoral neck, even after accounting for prolactin levels, breast-feeding status, return of menstruation, estradiol levels, PTH levels, 1,25 dihydroxyvitamin D levels, dietary calcium intake, physical activity, and body size. CONCLUSION: These data clearly support the hypothesis that PTHrP is an alternative mechanism associated with bone loss and recovery during and subsequent to lactation. PMID- 8709403 TI - Elevated plasma lipoprotein(a) and coronary heart disease in men aged 55 years and younger. A prospective study. AB - OBJECTIVE: To establish whether elevated lipoprotein(a) [Lp(a)], detected as a sinking pre-beta-lipoprotein band on electrophoresis of fresh plasma, is an independent risk factor for the development of premature coronary heart disease (CHD) in men. DESIGN AND SETTING: Prospective study of the Framingham offspring cohort. PARTICIPANTS: A total of 2191 men aged 20 to 54 years old who were free of cardiovascular disease when they were examined between 1971 and 1975. MAIN OUTCOME MEASURES: Incident CHD (myocardial infarction, coronary insufficiency, angina pectoris, or sudden cardiac death) occurring by age 55 years. RESULTS: After a median follow-up of 15.4 years, there were 129 CHD events. The relative risk (RR) estimates (with 95% confidence intervals [CIs]) for premature CHD derived from a proportional hazards model that included age, body mass index, and the dichotomized risk factor covariables elevated plasma Lp(a) level, total cholesterol level of 6.2 mmol/L (240 mg/dL) or more, high-density lipoprotein (HDL) level less than 0.9 mmol/L (35 mg/dL), smoking, glucose intolerance, and hypertension were as follows: elevated Lp(a) level, RR, 1.9 (95% CI, 1.2-2.9), prevalence, 11.3%; total cholesterol level of 6.2 mmol/L or more, RR, 1.8 (95% CI, 1.2-2.7), prevalence, 14.3%; HDL level of less than 0.9 mmol/L, RR, 1.8 (95% CI, 1.2-2.6), prevalence 19.2%; smoking, RR 3.6 (95% CI, 2.2-5.5), prevalence, 46.7%; glucose intolerance, RR, 2.7 (95% CI, 1.4-5.3), prevalence, 2.6%; hypertension, RR, 1.2 (95% CI, 0.8-1.8), prevalence, 26.3%. CONCLUSIONS: Elevated plasma Lp(a) is an independent risk factor for the development of premature CHD in men, comparable in magnitude and prevalence (ie, attributable risk) to a total cholesterol level of 6.2 mmol/L (240 mg/dL) or more, or an HDL level less than 0.9 mmol/L (35 mg/dL). PMID- 8709405 TI - Current trends in physicians' practice arrangements. From owners to employees. AB - OBJECTIVE: To examine current trends in practice organization among postresident patient care physicians in the United States. DESIGN AND SETTING: The American Medical Association's Socioeconomic Monitoring System (SMS), a series of periodic surveys of nationally representative samples of nonfederal postresident patient care physicians. Physicians were divided into 3 categories based on the organization of their main practice. They were classified as employee physicians if they had no ownership interest in their practice, as self-employed solo physicians if they were in 1-physician practices in which they had an ownership interest, and as self-employed group physicians if they were in multiple physician practices in which they had an ownership interest. PARTICIPANTS: Nonfederal, postresident patient care physicians who provided more than 47 000 responses to SMS surveys between 1983 and 1994. MAIN OUTCOME MEASURE: The proportion of nonfederal postresident patient care physicians who were employees between 1983 and 1994. RESULTS: Between 1983 and 1994, the proportion of patient care physicians practicing as employees rose from 24.2% to 42.3% (P<.001), the proportion self-employed in solo practices fell from 40.5% to 29.3% (P<.001), and the proportion self-employed in group practices fell from 35.3% to 28.4% (P<.001). Most of these changes occurred in the latter half of the 12-year period. These trends, which are evident in virtually every segment of the patient care physician population, are especially prominent among young physicians. The growing proportion of employee physicians is associated with increases in the earnings of employee physicians relative to those of self-employed solo physicians. CONCLUSIONS: Current trends in the US health care system are rapidly changing the career opportunities of patient care physicians and, hence, physicians' choice of practice arrangement. PMID- 8709406 TI - Niemann-Pick disease type C. From bench to bedside. PMID- 8709407 TI - Why sepsis trials fail. PMID- 8709408 TI - The growing conundrum. Growth hormone treatment of the non-growth hormone deficient child. PMID- 8709409 TI - Pharmacologic treatment of acute traumatic brain injury. PMID- 8709410 TI - A multi-institution collaborative policy on medical futility. PMID- 8709411 TI - [The 37th Congress of the Japanese Society of Clinical Hematology. Kyoto, Japan. October 23-25, 1995. Abstracts]. PMID- 8709412 TI - [38th Annual meeting of the Japan Geriatrics Society, Chiba, Japan. June 11-13, 1996. Abstracts]. PMID- 8709413 TI - [Effect of 1 alpha, 25-dihydroxy vitamin D3 on renal growth in rat]. AB - It has been demonstrated that 1,25 dihydroxy vitamin D3 [1,25 (OH)2D3] can control the growth of various cells in vitro. Despite the data obtained so far, is not known whether or not 1,25 (OH)2D3 can modulate renal growth. To determine the effect of 1,25(OH)2D3 on renal growth, 80 pmol of 1,25(OH)2D3 was administered intraperitoneally for one week consecutively to weanling Sprague Dawley rats using a minipump under the condition of strict pair-feeding. Kidney wet weight/body weight, renal protein/DNA ratio increased significantly, but renal RNA/DNA and DNA content were not affected. It was concluded that 1,25(OH)2D3 did promote renal growth in weaning rats, in contrast to the inhibitory effect of 1,25(OH)2D3 on renal cell proliferation as previously reported from earlier in vitro studies. Based on the evidence of an increased protein/DNA ratio, and unchanged RNA/DNA ratio and DNA content, renal growth seemed to have been the result of renal hypertrophy enhanced by 1,25(OH)2D3. PMID- 8709414 TI - [Angiographic study of stenosis and calcification of coronary vessels in long term dialysis patients: examination of risk factors for coronary calcification]. AB - Coronary atherosclerosis and calcification score were assessed angiographically in 30 uremic patients receiving dialysis (8 patients with diabetes mellitus [DM dialysis group] and 22 patients without DM [non-DM dialysis group] who were suspected of having coronary artery disease because of their clinical symptoms and electrocardiographic findings. Thirty non-uremic subjects (11 with DM and 19 without DM) served as controls. Twelve dialysis patients (40%) did not have significant coronary artery disease, and the rate of significant coronary artery disease in dialysis patients overall was less than that in the control subjects. The calcification score of dialysis patients was significantly higher than that of the control patients (P < 0.0001), and also scores of the non-DM dialysis group and the DM dialysis group were higher than those of each group of control subjects. The scores were significantly correlated with the duration of dialysis in uremic patients as a group (r = 0.52; P < 0.01) and the non-DM dialysis group (r = 0.70; P < 0.01), but were not correlated with the duration of dialysis in the DM dialysis group. The scores were correlated with serum phosphate concentration, but not with serum calcium concentration, calcium phosphate product, alkaline phosphatase levels total cholesterol or age. Calcification scores were extremely high (> 21) in six uremic patients who had high serum c terminal parathyroid hormone concentrations. These findings indicate that serum phosphate concentration, serum c-terminal parathyroid hormone concentration, and the duration of dialysis are closely associated with coronary calcification in long-term dialysis patients. PMID- 8709415 TI - [Plasma allantoin in patients undergoing maintenance hemodialysis]. AB - Recently, it has been noted that radical molecules participate in the pathogenesis of some of the complications associated with long-term dialysis therapy, such as amyloidosis. Since allantoin is produced from uric acid almost exclusively by the hydroxy radical, we investigated the plasma concentrations of allantoin in 71 patients with chronic renal failure who were on maintenance hemodialysis. Our specific objective was to investigate the relation of allantoin to the plasma concentrations of beta2-microglobulin and methylguanidine to evaluate the potential of allantoin as a significant parameter of the radical reaction. Although plasma allantoin was not detected in 15 healthy controls, plasma concentrations of allantoin increased markedly in all of the hemodialysis patients (42.6 +/- 37.7 nmol/mL) and wide variation was observed from 4.3 to 185.2 nmol/mL. In addition, we confirmed significant correlations between plasma concentrations of allantoin and beta2-microglobulin as well as serum concentrations of methylguanidine/creatinine and hyaluronic acid (r - 0.456, p < 0.0005, r = 0.313, p < 0.005, r = 0.368, p < 0.01, respectively). Although evidence for a direct link between plasma allantoin levels and amyloidosis was not obtained, the increase in the plasma concentrations of allantoin suggested its clinical significance as a parameter of the radical reaction that is thought to participate closely in the pathogenesis of dialysis-related amyloidosis. From a practical point of view, the measurement of plasma allantoin can be expected to become a significant method of measuring radical members which may be useful in the prophylaxis of some of the complications induced by free radicals. PMID- 8709416 TI - [Estimation of catecholamine release based on dialyzer clearance rate during hemodialysis]. AB - To estimate catecholamine (CA) release during hemodialysis (HD), plasma-free and conjugated CAs and their dialyzer clearance rates were measured in 10 HD patients (age; 49.8 +/- 15.2 years, duration of HD; 5.8 +/- 5.0 years). Although free dopamine (f-DA) and all conjugated CAs decreased to about one half of the pre-HD levels at the end of HD, no significant change was seen in free norepinephrine (f NE) and epinephrine (f-E) during HD. For every CA, the clearance rate was the highest in the sulfate and the lowest in the glucuronide form, and NE was the highest in every form. In the comparison between the measured CA and calculated CA using the clearance rate and the pre-HD level, the measured values of f-NE and f-E were significantly higher than the calculated values, unlike the results for f-DA and conjugated CAs. The difference in f-NE between the measured and the calculated values correlated negatively with the change in mean blood pressure (delta MBP), and delta MBP was also correlated with the ultrafiltration volume. From these data, it was suggested that f-NE was released by the decrease of MBP due to the increase of ultra-filtration volume during HD. PMID- 8709417 TI - [Clinical assessment of renal proximal tubular function using lithium clearance method]. AB - Proximal and distal sodium reabsorption values were calculated from lithium clearance in 63 patients with renal diseases, 13 cirrhotic patients with ascites and 12 control subjects. In the patients with renal diseases, fractional excretion of lithium (FELi) and fractional proximal reabsorption of sodium (FPRNa) were not changed in patients whose glomerular filtration rate (GFR), was over 30 mL/min, but FELi was increased and FPRNa was decreased when the GFR was lower than 30 mL/min. Moreover, fractional distal reabsorption of sodium (FDRNa) was decreased in patients whose GFR was under 40 mL/min. These results indicate that proximal tubular function is well adapted to the degree of renal function even if the etiologies of renal diseases are different. Five patients with nephrotic syndrome (minimal change type) were subjected to lithium clearance method before and after steroid treatment. FPRNa in nephrotic patients was reduced after the treatment, though there was no significant difference in FDRNa. In cirrhotic patients, FELi, FPRNa and FDRNa did not differ from the values in the control subjects, which were not influenced by the decrease in GFR. Thus, the reduction of FPRNa with GFR which was observed in renal disease, was absent in liver cirrhosis. In conclusion, these data indicate that renal adjustment of sodium excretion in chronic renal disease at first takes place in the distal parts of the nephron and later in the proximal tubule, and in addition, that in appropriate reabsorption of sodium from the proximal tubule probably plays a role in ascites formation in cirrhotic patients. PMID- 8709418 TI - [A case of selective IgM deficiency associated with systemic lupus erythematosus]. AB - We report a case of selective IgM deficiency associated with systemic lupus erythematosus (SLE). A 34-year-old female suffering from SLE was admitted with proteinuria and general fatigue. Laboratory findings revealed a very low serum IgM level, almost lower than 12 mg/dl. Renal biopsy findings showed diffuse proliferative lupus nephritis (DPLN). In immunofluorescent microscopy, IgG was the most strongly stained followed by IgA, but IgM staining was only faint. As for the immunophenotype of the T cells, the OKT4/OKT8 ratio was normal. Response to both phytohemagglutinin (PHA) and concanavalin A (ConA) was normal. However, responses of B cells to both pokeweed mitogen (PWM) and Staphylococcus aureus Cowan strain I (SAC) were significantly reduced. Surface IgM-positive B cells were decreased. These results indicate that the patient had B cell dysfunction, involving impairment of B cell differentiation. In this report, we discuss selective IgM deficiency and SLE documented in the literature. PMID- 8709419 TI - [A case of resected small-sized (4 mm in diameter) lung cancer and its way of surgical treatment]. AB - We report a case of resected lung cancer which was 4 mm in diameter. Pathological diagnosis couldn't be made preoperatively by fiberoptic bronchoscopy. We examined the patient's chest CT carefully and determined the presence or absence of such findings that might indicate malignancy (spicula formation, involvement of pulmonary vasculature, irregularity of tumor edge, pleural indentation, and notching) or benignity (central or diffuse calcification, sattelite lesions). When a lesion shows more than two of five findings listed above that indicate malignancy, and when the diagnosis of lung cancer is agreed upon by a team consists of a radiologist, a physician and a surgeon, we would tell the patient and his/her family that the lesion is most likely a lung cancer, and recommend surgical treatment. Sometimes small-sized lung cancer isn't a early lung cancer so recommend curative standard operation if patient's condition is good. PMID- 8709420 TI - [Clinical experience of coronary artery bypass grafting using surgical microscope]. AB - Since 1991, we have attempted to use a surgical microscope for 62 coronary bypass anastomoses. Four kinds of microscopes were used under high magnification (6 to 8x). The suturing thread length was 5-10 cm to make anastomosis and handling of the needle easier. It was concluded that a surgical microscope could be best used for anastomosis of an internal thoracic artery graft with the left anterior descending coronary artery. Excellent graft patency and improvement of the quality of anastomosis were confirmed. PMID- 8709421 TI - [A case of pulmonary dirofilariasis diagnosed through thoracoscopic lung biopsy]. AB - A fifty-year-old man was admitted to our hospital for evaluation of an abnormal shadow in a routine chest X-ray film. Thorax CT scan revealed a relatively well defined nodule with a diameter 15 mm and pleural thickness in the posterior segment of the right lower lobe. A diagnosis by broncoscopy was undefinite and suspected to be a lung cancer. So thoracoscopic lung biopsy was performed to remove it. In a cross section of the resected nodule two degenerated worm structures were revealed and we diagnosed pulmonary dirofilariasis. Pulmonary dirofilariasis has been an increasing reported disease. Most of cases revealed a peripheral lung lesion. Thoracoscopic lung resection was useful for the diagnosis, considering as minimally invasive examination. PMID- 8709422 TI - [Anticoagulant related hemorrhage and thromboembolism after valvular surgery]. AB - We reviewed anticoagulant related hemorrhage (ACRH) and thromboembolism (TE) in 84 patients after valvular surgery. There were 45 females and 39 males with a mean age of 51.8 years (range 30.5-71.2 years), who underwent valvuloplasty in 14, bioprosthetic valve replacements in 17, mechanical valve replacements in 13. A mean period from the operation to the event were 2.7 years (range 0.01-12.3 years). There were 25 ACRH events after one valvuloplasty, 4 bioprosthetic valve replacements, 20 mechanical valve replacements. About half of them, the prothrombin time were less than 25%, which was considered the effect of warfarin is high, and 8% of them had infective endocarditis (IE) previously. There were 59 TE events after 13 valvuloplasties, 13 bioprosthetic valve replacements, 33 mechanical valve replacements. In the patients with atrial fibrillation, TE occurred irrespective of operative procedures. And in the patients with mechanical valve, severely impaired left ventricular function and past history of IE, thrombi of left ventricule were caused of TE. It was suggested that past history of IE was a risk factor ACRH and TE, and severely depressed left ventricular function and atrial fibrillation were for TE. PMID- 8709423 TI - [Hemodynamic plus series of St. Jude medical valve prosthesis in aortic position: early clinical outcome and reconsideration of its application]. AB - St. Jude Medical Hemodynamic plus bileaflet valve prosthesis (HP) was employed in 7 cases undergoing aortic valve replacement since December 1993. Echocardiographic evaluation of left ventricular dimension and transvalvular pressure gradient was performed in 5 cases with 19 HP before and after operation. Each data was compared with those of 21 cases having undergone aortic valve replacement with 19 mm Standard model (19 SD). There was no significant postoperative change in left ventricular end-diastolic dimension (50 +/- 4 mm to 44 +/- 8 mm; p = NS) and left ventricular end-systolic dimension (34 +/- 4 mm to 34 +/- 9 mm; p = NS) in 19 HP cases. However, left ventricular end-diastolic dimension (51 +/- 11 mm to 41 +/- 8 mm; p < 0.05) and left ventricular end systolic dimension (35 +/- 12 mm to 28 +/- 9 mm; p < 0.05) decreased in 19 SD cases. Transvalvular systolic peak pressure gradient was reduced significantly in both groups (19 HP: 94 +/- 38 mmHg to 43 +/- 19 mmHg; p < 0.05, 19 SD: 73 +/- 49 mmHg to 33 +/- 14 mmHg; p < 0.05). Poor reduction of left ventricular volume and high residual systolic pressure gradient in patients with 19 HP might be due to relatively high incidence of stenotic lesion in original aortic valve disease. In some cases with these critical aortic stenosis, radical annular enlargement procedure would provide more fundamental release of left ventricular outflow tract stenosis rather than one size increase of effective orifice area by employing HPs. Structural examination demonstrated that 19 HP had the same outer diameter of sewing flange with identical orifice ring as 21 SD (24 mm) did, and 17 HP had that of 19 SD (22 mm). Therefore, precise attention is needed for application of SJM-19 HP in patients with small aortic annuli. PMID- 8709424 TI - [Nine cases of spontaneous hemopneumothorax with massive bleeding]. AB - We experienced nine cases of spontaneous hemopneumothorax. All the patients were male and six of those had received emergent operations because of massive intrathoracic bleeding. In six of the nine cases we found the bleeding from the ruptured string like tissues between parietal pleura and the apex of the upper lobe. Those string like tissues contained vessels and were suspected to have ruptured when the lung collapsed. In other three cases the bleeding was controlled conservatively but operations were performed because of continuous air leakage and the bleeding were found from the bullous apical tissues of the lung adhered to the thoracic wall. Histopathologically those bullous tissues and string like tissues were revealed grannuromatous and chronic pleuritis rich in microvessels. Early thoracotomy for spontaneous hemopneumothorax was recommended in case of massive intrathoracic bleeding. PMID- 8709425 TI - [Iatrogenic injury of tracheobronchial membranous wall]. AB - Tracheobronchial injuries remain uncommon, but they are of great significance, because they can result in death or substantinal functional compromise. Such injuries are mostly from blunt trauma and motor vehicle accidents, but there also is an incidence of penetrating thoracic trauma inclunding iatrogenic accidents. Three cases of iatrogenic injury of tracheobronchial membranous wall were reported. Two cases were tear at the membraneous portion of the left main bronchus and trachea by forceful endotracheal intubation. Another case was tracheal membranous wall injury during operation at blunt dissection for esophageal carcinoma. We reported the emergent managements for iatrogenic injury of tracheobroncheal membranous wall in differents 3 ways. We should select the best treatment according to the condition of the patients and situation of the injury. PMID- 8709426 TI - [Surgical treatment for ruptured aneurysm of distal arch or aortic arch: report of three cases]. AB - We experienced three cases of surgery for ruptured aneurysm of the distal aortic arch (2 cases) or aortic arch (1 case). A 59-year-old man who was diagnosed as distal arch aneurysm ruptured to the pericardial cavity underwent graft replacement of the aneurysm and reconstruction of a left subclavian artery under selective cerebral perfusion (SCP). This patient had left hemiplegia postoperatively. A 73-year-old man who was diagnosed as an aneurysm of distal arch and descending aorta ruptured to left pleural cavity underwent graft replacement of the aneurysm and reconstruction of a left subclavian artery under SCP, and died of severe left ventricular failure possibly due to preoperative myocardial ischemia. A 70-year-old man who was diagnosed as a saccular aneurysm of aortic arch ruptured to mediastinum underwent resection of the aneurysm and patch plasty of the aorta under SCP. The postoperative course was uneventful. We discussed the approach to aneurysm, adjunctive method and surgical precedures from standpoint of our ruptured three cases. PMID- 8709427 TI - [Surgical results of total replacement of aortic root]. AB - Total aortic root replacement was performed on 19 cases since December 1979 to May 1995. They were devided in two groups. First 7 cases were operated by original Bentall procedure with inclusion technique (group A). Succeeding 12 cases were operated by Carrel patch method or Cabrol procedure without inclusion maneuver. There were 2 operative deaths in group A (28.6%) and was one death in group B (8.3%) respectively. The operation time and the extracorporeal circulation time were significantly shorter in group A than that in group B. Anastomotic leakage was noted in 2 cases of group A. One of them required reoperation and another one with coronary ostial leakage died of congestive heart failure. We concluded that non-inclusion method for aortic root replacement was superior to conventional inclusion technique and reimplantation of both coronary ostia with Carrel patch method was the best tactics especially for Marfan syndrome in terms of long term results. PMID- 8709428 TI - [Coronary artery bypass grafting for ischemic heart disease associated with acromegaly: a case report]. AB - Coronary artery bypass grafting (CABG) for ischemic heart disease associated with acromegaly has never been reported in Japan, though coronary artery sclerosis is known as complicating factor in acromegaly. A 44-year-old man with ischemic heart disease was admitted to our hospital and emergency percutaneous transluminal coronary recanalization was performed. After admission, he was diagnosed as acromegaly from endocrinological studies and operated on CABG. The peri- and post operative course was uneventful in spite of our fear for complications due to acromegalic heart muscle disease. PMID- 8709429 TI - [An operative case of bilateral coronary arteries fistulae to pulmonary artery associated with a giant saccular aneurysm]. AB - We reported an operative case of bilateral coronary artery fistulae to pulmonary artery associated with a giant saccular aneurysm, the largest of which measured 30 x 30 mm. The patient was a 75 year old female who had anginal pain due to coronary steal phenomenon. A continuous murmur was detected. After establishing total cardiopulmonary bypass, two distal orifices of the fistula connected to the main pulmonary artery were closed with 5-0 polypropylene plegeted sutures. Aneurysmorrhaphy was then performed for giant saccular aneurysm. Postoperative course was uneventful. PMID- 8709430 TI - [Endovascular stenting for a recurrent angina patient]. AB - We report successful Palmaz-Schatz stenting for recurrent angina. Coronary arteriography in a 65-year-old man with recurrent angina six months after coronary artery bypass grafting (CABG) showed stenosis in the right coronary artery and in the saphenous vein graft to the left anterior descending artery. For both stenotic lesions, balloon dilatation was performed. However, both lesions recoiled after dilatation. Therefore, Palmaz-Schatz stents were inserted in both stenotic lesions. After stenting, the patient did not experience any further chest pain. Three months later coronary arteriography showed the stents remained patient. Balloon angioplasty for post-CABG angina is technically easier than additional surgery. However, angioplasty by balloon dilatation alone can induce dangerous complications during the acute phase and restenosis frequently develops. This method is not sufficiently effective or safe for recurrent angina. To prevent these problems, the stent was developed as a useful adjunct to balloon dilatation for the native artery. Using this new device, angioplasty for recurrent angina will be safer and more effective than additional surgery. PMID- 8709431 TI - [A case report of mitral valve plasty for traumatic mitral regurgitation]. AB - A 64-year-old man was admitted to our hospital with a chief complaint of general fatigue on June 24, 1993. He had no history of rheumatic fever but bruised in the chest in September 1990. After that, his systolic murmur was noticed at the apex and echocardiogram showed the posterior chordal rupture of mitral valve. Mitral valve plasty was performed successfully. PMID- 8709433 TI - [Supraannular implantation of bioprosthetic valve for severe tricuspid valve regurgitation associated with atrial septal defect in an adult patient]. AB - With an adult atrial septal defect, there was often regurgitation of the mitral and tricuspid valve due to volume overload in a long term period. Especially concerning tricuspid regurgitation, what can be done surgically has not yet been decided. For severe tricuspid regurgitation, some cases where tricuspid valve annuloplasty were performed have had exacerbation of tricuspid regurgitation. We experienced a case where tricuspid valve supraanular implantation without excision of native tricuspid valve (TVSI) was performed for severe tricuspid regurgitation associated with atrial septal defect, and improved. A 53-year-old female complained of dyspnea on exertion. An atrial septal defect was revealed being 4 cm in size, complicated with severe tricuspid regurgitation (IV), and 62 mmHg difference of pressure from the right atrium to right ventricle shown by a ultrasonography. Pulmonary artery pressure was 66/16 mmHg by cardiac catheter. Patch closure for ASD and TVSI for TR was performed on her, and amelioration of cardiac function was recognized. PMID- 8709432 TI - [Surgical treatment of double-orifice mitral valve associated with incomplete type endocardial cushion defect]. AB - We report surgical treatment of double-orifice mitral valve (DOMV) associated with incomplete-type endocardial cushion defect. All three cases had "central type DOMV classified by Cascos" with bridging tissue between two mitral orifices. We closed mitral cleft in the two cases with moderate mitral regurgitation, although we left the bridging tissue. It is thought to be beneficial to leave the bridging tissue undivided to avoid postoperative mitral regurgitation in such cases. PMID- 8709434 TI - [A surgically treated case of ruptured true aortic arch aneurysm in association with Stanford type A chronic dissection]. AB - A 72-year-old woman was admitted to our hospital because of hemosputum. Enhanced CT showed ruptured true aortic arch aneurysm. True aortic arch aneurysm ruptured at distal portion of aneurysm. The aorta was chronically dissected from an entry proximal to ruptured portion, to ascending aorta. Ascending aorta to aortic arch was replaced with 20 mm gelatin sealed graft under selective cerebral perfusion with hypothermia. Post-operatively tracheostomy was done because of respiratory failure, and there was some leak from distal anastomosis portion which was 42 mm in diameter at the operation. So replacement of descending thoracic aorta was performed using elephant trunk of the previously replaced graft. Post-reoperative course has been uneventful. She weaned from respiratory support. True aortic arch aneurysm combined with Stanford type A dissection is very rare. We presented ruptured true aortic arch aneurysm in association with Stanford type A chronic dissection which had an entry in true aneurysm. PMID- 8709435 TI - [A case report of St. Jude medical aortic valve prosthesis thrombosis]. AB - We report a case of successful semi-emergent redo aortic valve replacement for thrombosed prosthetic valve. Patient was a 59-year-old male who had undergone aortic valve replacement of St. Jude Medical prosthetic valve and open mitral commissurotomy 14 years ago. He suffered from pneumonia with hemosputum and admitted a neighbor hospital. But he was reffered to our medical center because of progressing dyspnea. We found the immobility of one of the leaflets of the prosthetic valve by cinefuluoroscopy. He was also diagnosed as prosthetic valve thrombosis and mitral valve restenosis using ultrasonic echocardiography. The thrombi were developed on the hinge of one of the leaflets, and they might be formed during the time he had got inappropriate anticoagulation therapy at the former hospital. We emphasise the importance of strict anticoagulation control for the patients valve replaced. PMID- 8709437 TI - [Experience with 2 cases of traumatic diaphragmatic hernia occurred at the same time]. AB - Two cases of traumatic diaphragmatic hernia (TDH) which occurred at the same traffic accident, are reported. The first case, a plain chest X-P was diagnostic of TDH. With thoracotomy and additional laparotomy, lacerated diaphragm was repaired. The second case, continual observation on plain chest X-P was useful to suspect TDH, and thoracoscopy was diagnostic. With thoracotomy diaphragm was repaired. Two patients recovered well. It has not been reported that two cases of TDH occurred at same trauma. For diagnosis of TDH, chest X-P is useful for suspicion, and thoracoscopy is useful for correct diagnosis. Surgical approach should be decided in consideration of phase, location, and complicated injuries. PMID- 8709436 TI - [A case of surgically treated hypertrophic obstructive cardiomyopathy suffering from angina pectoris during hemodialysis]. AB - Surgical procedure for hypertrophic obstructive cardiomyopathy (HOCM) in a patient chronic hemodialysis has not been reported in Japan. This is a case report of a 61-year-old male who had been suffering from angina pectoris during hemodialysis. Cardiac catheterization showed a systolic pressure gradient of 79 mmHg at the left ventricular outflow tract and left ventriculogram demonstrated severe mitral regurgitation. He underwent mitral valve replacement and left ventricular myectomy with hemodialysis and extracorporeal ultrafiltration method during cardiopulmonary bypass. Postoperative catheterization showed a decrease of pressure gradient to 19 mmHg at the left ventricular outflow tract, and relief of angina was obtained. PMID- 8709438 TI - [A case of tracheal stenosis caused by invasion of recurrent lung cancer palliated with dumon stent after Nd-YAG laser and bouginage using of tracheal tube]. AB - The patient was 67 year-old male with tracheal stenosis caused by invasion of recurrent lung cancer after right upper sleeve lobectomy. He visited to our hospital for the purpose of palliation with tracheal stent. But rigid bronchoscope cannot be passed through its lesion because of severe stenosis. So the bouginage was performed. This method was that tracheal tube was inserted into his trachea through the hole of tracheotomy during six days following Nd-YAG laser therapy. After this treatment, rigid bronchoscope was inserted into the trachea and Dumon stent was placed safely and easily. It seems to be an useful palliation for patients with severs invasive tracheal stenosis. PMID- 8709439 TI - [Cellular biological approach for determining of malignant potential and invasiveness of bladder cancers]. PMID- 8709440 TI - [Ectopic production of HCG beta by bladder carcinoma in vitro and in vivo]. AB - BACKGROUND: Ectopic production of immunoreactive hCG/hCG beta (IR-hCG beta) by bladder tansitional cell carcinoma cell lines was investigated in vitro and in vivo. METHODS: As an in vitro study, IR-hCG beta in culture media from 2 bladder transitional cell carcinoma cell lines (KoTCC-1 and HT-1197) was analyzed by three kinds of enzyme immunoassays (EIA) which were specific for intact hCG, free hCG beta and beta-core fragment (beta-CF). As an in vivo study, distribution of IR-hCG beta was analyzed in tumor tissues, sera, and urine of the nude mice and the nude rat transplanted with KoTCC-1 cell line. RESULTS: Both of the cell lines were determined to secrete IR-hCG beta into the media, which consisted principally of free hCG beta. Intact hCG and beta-CF were scarecely detected in the media. Immunohistochemical study revealed the localization of IR-hCG beta in transitional cell carcinoma cells of the transplanted tumor. Although a large amount of IR-hCG beta could be detected in both of the serum and urine from the animals, there were quantitative and qualitative differences between serum and urinary IR-hCG beta. Quantitatively, the concentrations of IR-hCG beta in the urine were consistently much higher than those in the serum. Qualitatively, free hCG beta was exclusively detected in the serum whereas a large amount of beta-CF, in addition to free hCG beta, were found in the urine. Intact hCG could not be detected in both serum and urine. These distributions of IR-hCG beta in the animals bearing tumors were completely analogous to those in patients with bladder carcinoma. CONCLUSION: The present results suggested that ectopic production of IR-hCG beta by bladder carcinoma is not rare phenomenon and it is clinically useful as a tumor marker when beta-CF is measured in the urine. PMID- 8709441 TI - [A basic study of the interstitial laser prostatectomy using pulsed holmium: yttrium-aluminum-garnet (Ho:YAG) laser]. AB - BACKGROUND: To develop a new procedure for laser prostatectomy through a biopsy needle, we evaluated the efficacy of interstitial laser irradiation with a pulsed Ho:YAG laser (lambda: 2.1 microns) in the normal canine prostate. METHODS: Pathological studies were performed up to 6 months after interstitial laser irradiation in 26 mongrel dogs (15.4 +/- 2.6 kg). Two dogs underwent the identical procedures without irradiation as controls (control group). Prostate glands of the other 24 dogs were irradiated interstitially with a Ho:YAG laser through a silica glass fiber (400 microns phi). In one prostate, each lobe was irradiated with 500 J/cm2 or 150 J/cm2 in energy density (fluence). Four prostates were irradiated with 150-175 J/cm2 in fluence (low-fluence group). Nineteen prostates were irradiated with 500-600 J/cm2 in fluence (high-fluence group). Prostates glands were excised immediately, 1 month, 2 months, 3 months, or 6 months after irradiation and examined histologically. To investigate the influence of interstitial irradiation with a Ho:YAG laser on smooth muscle and/or on the adrenergic activity of the prostate, we also measured changes in intraluminal pressure of the prostatic urethra upon administration of epinephrine (10 micrograms/kg) in six dogs before and after irradiation. We developed a novel three-membrane balloon catheter with an open end for measuring this pressure change. RESULTS: Specimens harvested immediately in control and low-fluence groups showed slight hemorrhage in areas surrounding the punctured or irradiated site, but the surrounding smooth muscles or small vessels were not destroyed. In the high-fluence group, smooth muscles and small vessels approximately 1 mm thick that surrounded the ablated hole were destroyed, but the thickness of the thermal coagulation layer was only approximately 0.2 mm. These findings suggested that damage to the small vessels and smooth muscle may have been caused by laser induced stress waves rather than by a thermal effect. In the control and low fluence groups, prostatic volume remained unchanged 1 month after puncture or low fluence irradiation. In the high-fluence group, huge cavities were created in the laser-irradiated prostate gland 1 or 2 months after irradiation and prostatic volume was reduced maximally by 60%. In the high-fluence group, the cavities collapsed and prostatic volume was reduced by up to 50% at 6 months post procedure. The prostatic urethra expanded following the reduction in volume. The maximal intraluminal change in pressure of the prostatic urethra decreased significantly after laser irradiation (p < 0.05). CONCLUSION: Results indicate that interstitial irradiation of the prostate with a Ho:YAG laser with high fluence may relieve the anatomical and functional obstruction of the prostatic urethra in benign prostatic hyperplasia. PMID- 8709442 TI - [Clinicopathological characteristics of small size renal cell carcinoma]. AB - BACKGROUND: We tried to identify which size or less means small size in renal cell carcinoma (RCC) associated with prognosis including Robson's stage 1. METHODS: We classified RCC based upon the tumour size into cases with 2.5 cm or less (NO, VO, MO: group I: n = 27, cause-specific cases = 26) and cases with 2.6 to 3.0 cm (NO, VO, MO: group II: n = 31, cause-specific cases = 27). Based upon this classification, we studied on the clinical features of small size RCC. RESULTS: (1) Regarding the clinical features including mean age, sex ratio, affected side, opportunity of diagnosis, diagnostic approach and preoperative laboratory findings, no specific difference was observed between groups I and II except for the localization of tumour in the kidney (a significantly high rate was observed in the mid portion of the tumour in group I compared with that in group II). 2) The mean size of group I was 1.98 cm (0.7 to 2.5 cm), and that of group II was 2.84 cm (2.6 to 3.0 cm). On the other hand, the mean size of Robson's stage 1 was 5.5 cm (0.7 to 18.0 cm). Therefore, the tumour size of Robson's stage 1 was significantly larger than that of groups I and II (p < 0.001). 3) The survival rate of group I showed worse trend than that of group II after 12 years postoperatively, but no significant difference was observed between these 2 groups. Furthermore, the survival rate of Robson's stage 1 showed better trend than that of the tumour size with 3.0 cm or less, but no significant difference was observed between these 2 groups. 4) As to the grade in each group, no significant difference of survival was observed. 5) As to the recurrent rate in each group, it was 19.2% in group I, 7.4% in group II and 18.5% in Robson's stage 1. Therefore, no significant difference of recurrent rate was observed in these groups. CONCLUSION: It is difficult to identify which size or less means small size RCC associated with prognosis. This is because no specific clinical feature was observed in these small size cases compared with Robson's stage 1 which consisted with larger size tumour. PMID- 8709443 TI - [A morphological study of renal adenoma and latent renal cell carcinoma in autopsy cases]. AB - PURPOSE: We investigated histopathological features of renal adenoma and latent renal cell carcinoma in order to find relationship between these two entities. MATERIALS AND METHODS: Renal adenomas or latent renal cell carcinomas found macro and microscopically in 2,201 autopsy materials were used. Histological examination was focused on the size, and the presence or absence of hemorrhage, necrosis and capsulation of tumors. Additionally, we analyzed morphologically the mean nuclear area (MNA), N/C ratio, mean nuclear volume (MNV) and nuclear roundness factor (NRF) using image analyzer system (CIA-10; Olympus). RESULTS: A total of 71 foci of renal cortical tumor was found and classified into renal adenoma (59 foci), latent renal cell carcinoma (8 foci), and intermediate type (4 foci). Intermediate type contained of small element of clear cell and were difficult to distinguish from renal cell carcinoma or ordinary adenoma. The mean diameter of renal adenomas was 1.9 mm. The capsulation was found in only 3 foci (5.1%) of them, and bleeding and necrosis were found in none of them. On the other hand, the mean diameter of latent renal cell carcinoma and intermediate type were 15.7 mm and 6.6 mm, respectively, and larger than that of renal adenomas (p < 0.005). In latent renal cell carcinoma, capsulation were found in all foci, bleeding in 6 (75.0%) and necrosis in 3 (37.5%), and their incidence were significantly higher than those of adenoma (p < 0.005). In intermediate type, capsule and bleeding were found in each two foci. Morphologically, there were significant difference in MNV and NRF but no difference in MNA and N/C ratio between renal adenomas and latent renal cell carcinomas. Intermediate type was larger in NRF compared to adenoma. CONCLUSION: Morphological characteristics of intermediate type resemble that of latent renal cell carcinoma in terms of tumor size, the presence of bleeding and capsule and NRF when compared to adenomas. These results suggest that, in some renal cell carcinoma, adenoma might be able to change to carcinoma. PMID- 8709444 TI - [En bloc cystourethrectomy with preservation of potency]. AB - BACKGROUND: The objective of this study is to evaluate efficacy of en bloc cystourethrectomy with preservation of potency in patients with bladder carcinoma. METHODS: A total of nine patients underwent this procedure in 1991 and 1994. Following 2 cases of 1991 who were confirmed to recover potency and had no recurrence in the pelvis during 2-year follow up as a preliminary study, 7 cases underwent the same procedure in 1994. Erectile function was measured by the snap gauge band. RESULTS: Six of the 9 patients recovered erection adequate for sexual activity and 4 of them have practiced sexual intercourse. The remaining 3 patients experienced mild erection but inadequate. One of them failed to accomplish adequate erection because of perineal pain at erection. Three to eighteen months after surgery, the first erection was experienced. Surgical margins were pathologically negative in each of the 9 specimens. By the present, all of the 9 patients survive with no evidence of disease. CONCLUSION: The potency-sparing en bloc cystourethrectomy appears to preserve potency on the level comparable to the two-stage potency-sparing cystourethrectomy combined with delayed urethrectomy. PMID- 8709445 TI - [Intermittent intra-arterial chemotherapy for the treatment of patients with bladder cancer or prostate cancer using implantable injection pump]. AB - BACKGROUND: The objective of this study is to evaluate the usefulness of intra arterial chemotherapy using an implantable injection system for the treatment of bladder or prostate cancer. METHODS: Twenty-four patients with bladder cancer and 13 with prostate cancer were treated with intermittent intra-arterial chemotherapy using a totally implantable injection pump. A heparinized catheter was percutaneously fixed through the femoral artery with the tip 2 or 3 cm above the bifurcation of the common iliac artery, and the pump was placed in a subcutaneous pocket. RESULTS: The total number of infusion sessions for bladder cancer ranged from 4 to 38 times (mean 20.8), and 15 of the 24 patients were injected from 2 to 19 sessions (mean 8.5) as outpatients. The tumor response for 15 patients with newly diagnosis muscle-invasive bladder cancer were 12 patients in CR, 1 in PR, and 2 in NC, and 9 patients with recurrent bladder cancer were 6 patients in CR, 1 in PR, and 2 in NC. The total number of infusions for prostate cancer ranged from 6 to 35 times (mean 20.2), and 10 of the 13 patients were injected from 1 to 11 times (mean 6.3) as outpatients. All 13 patients with prostate cancer were treated with hormone therapy and irradiation, and achieved PR. Complications of this drug delivery system were obstruction of the catheter or pump observed in 7 patients, dislocation of the catheter in 2, and obstruction of right external iliac artery in 1. CONCLUSION: These findings suggest that favorable therapeutic efficacies and an improved quality of life for patients can be obtained by intra-arterial chemotherapy using an implantable injection pump. PMID- 8709446 TI - [Prognostic factors in patients with advanced prostatic cancer (stage D2) initially treated with endocrine therapy]. AB - BACKGROUND: In order to make clear the prognostic factors in patients with advanced prostatic cancer (stage D2) whose initial treatment was hormone therapy, we reviewed 34 men with newly diagnosed advanced prostatic cancer for 6 years from May 1987 at the Hospital of Hokkaido University. METHODS: All patients had histologically proven prostatic adenocarcinoma. The mean age was 67.9 years (range 54 to 86 years). Median follow-up period was 26 months (range 5 to 66 months). The parameters studied were age, performance status, histological grade, extent of disease on bone scan, pretreatment values of PSA, gamma-Sm, PAP and PSA/gamma-Sm ratio, posttreatment values of PSA, gamma-Sm and PAP at 6th month, and the ratio of PSA at 3rd month to PSA at 6th month. RESULTS: Univariate analysis revealed that performance status (PSO vs. PS2, p = 0.006, PS1 vs. PS2, p = 0.016) and extent of disease on bone scan (p = 0.004) in background factors, gamma-Sm (p = 0.005) among pretreatment values of markers, PAP at 6th month (p < 0.001) and the ratio of PSA at 3rd month to PSA at 6th month (p < 0.001) among posttreatment values of markers, significantly related with the cause-specific survival. Based on multivariable analysis, PAP at 6th month and the ratio of PSA at 3rd month to PSA at 6th month were significantly correlated with the cause specific survival at 24th month and 36th month, respectively. CONCLUSION: We conclude that posttreatment values of markers are more important prognostic factors in patients with advanced prostatic cancer (stage D2) following hormone therapy than pretreatment background factors and values of markers. PMID- 8709447 TI - [Relationship between the quantitative morphometry and the clinical response to alpha adrenergic blocker in benign prostatic hyperplasia]. AB - BACKGROUND: There have been few reports on the relationship between the prostatic tissue components and the effect of alpha adrenergic blocker on urination. The present study was undertaken to correlate the rates of the prostatic tissue components and the clinical response to alpha adrenergic blocker in benign prostatic hyperplasia. METHODS: 40 patients with symptomatic benign prostatic hyperplasia were treated with 0.2 mg/day tamsulosin, a alpha adrenergic blocker, for 4-8 weeks followed by transurethral resection of the prostate. The mean weight of resected prostatic tissues was 25.2 g (range 5-96 g). Specimens were stained with antibodies to muscle-specific actin and to prostate-specific antigen for the detection of the smooth muscle and the glandular epithelium, respectively. Double-immunoenzymatic stained tissue sections were divided into four groups: smooth muscle, connective tissue, glandular epithelium, and glandular lumen. The rates of the areas of respective prostatic tissue components were evaluated with computer-assisted quantitative color image analysis. RESULTS: The mean rate of the area of smooth muscle, connective tissue, glandular epithelium, and glandular lumen was 32.7%, 46.9%, 10.8%, and 9.6%, respectively. International prostatic symptom score, maximum flow rate and residual urine volume before the treatment were not related to the rates of respective prostatic tissue components. International prostatic symptom score and residual urine volume were reduced and maximum flow rate was increased after the treatment with tamsulosin. A significant relationship was proved between the rate of smooth muscle and the change in maximum flow rate. The large hyperplastic adenoma contained less smooth muscle and showed a lower sensitivity to alpha adrenergic blocker than the small one. CONCLUSION: The clinical response to alpha adrenergic blocker was related to the rate of smooth muscle in benign prostatic hyperplasia. PMID- 8709448 TI - [Detection of prostate cancer in urological practice: clinical establishment of serum PSA reference values by age]. AB - BACKGROUND: Determinations of serum levels of prostate specific antigen (PSA) are widely used for the detection of prostate cancer, but have not demonstrated sufficient sensitivity and specificity to be useful in urological practice. In order to enhance the diagnostic value of serum PSA, we studied the distribution of serum PSA levels by age in urological patients without clinical evidence of prostate cancer and determined clinical PSA reference values by age decades. METHODS: The study included a total of 590 male patients aged 40 to 89 years who visited our urological clinic complaining of voiding symptoms from January 1991 to October 1994, but had no clinically evident prostate cancer. We defined patients to be without clinical evidence of prostate cancer if they had negative test results in digital rectal examination, ultrasonography, and serum PSA assay, or had positive test results in one or more of these tests but had a nonmalignant prostate biopsy. Serum PSA levels were measured by E-Test Tosoh II (AIA-PACK PA, normal range; 0 to 5.3 ng/ml). The patients were grouped by age decades and serum PSA values were given as percentiles. RESULTS: Analyzed by Pearson's correlation coefficient, serum PSA levels were correlated significantly with patient age (r = 0.24, p < 0.001). Prostatic volume was correlated weakly but significantly with patient age (r = 0.12, p = 0.005), and PSA density was also correlated significantly with patient age (r = 0.20, p < 0.001). Thus, serum PSA levels were demonstrated to increase with advancing patient age. Factors other than benign prostatic hypertrophy were also suggested to explain the increase in serum PSA levels in older patients. With the 95th percentile for serum PSA as the upper limit, the clinical PSA reference values by age decades were determined to be 2.6 ng/ml for patients aged 40 to 49 years, 5.0 ng/ml for 50 to 59 years, 7.5 ng/ml for 60 to 69 years, 10.1 ng/ml for 70 to 79 years, and 12.4 ng/ml for 80 to 89 years. CONCLUSION: We found a significant increase in serum PSA levels with advancing patient age. Thus, it is appropriate to have serum PSA reference values by age decades. Prospective clinical trials are necessary to define the usefulness of the PSA reference values by patient age in urological practice. PMID- 8709449 TI - [Renal metastasis originating from liver cancer]. AB - The patient was a 52-year-old man. He was suspected of having right renal carcinoma with hepatic metastasis and was referred to our hospital. The result of percutaneous needle biopsy on the right renal tumor showed a renal cell carcinoma. For the treatment, he underwent transcatheter arterial embolization (TAE) and intramuscular injection of interferon alpha. However, he was getting weaker and weaker and two months later he died due to hepatic failure. The result of autopsy showed that the renal carcinoma was a metastatic tumor from the primary hepatic cancer. This case was thought to be the second case of metastatic renal tumor from hepatic cancer reported in the Japanese literature. PMID- 8709450 TI - [A case of drug-induced immune hemolytic anemia during alpha-interferon therapy for renal cell carcinoma]. AB - A Case of drug-induced immune hemolytic anemia during alpha-interferon (alpha IFN) therapy for renal cell carcinoma is reported. A 61-year-old woman was admitted to Tochigi cancer center for the treatment of left renal cell carcinoma. She underwent left radical nephrectomy. From 7th post operative day, alpha-IFN (6 x 10(6) IU) was administered every other day. Diclofenac and indomethacin were administered for pain and high fever induced by alpha-IFN. Hemoglobinuria was first noted on 20th post operative day. Immune hemolytic anemia was suspected by blood examinations including Coombs' test, serum haptoglobin, serum LDH and serum GOT. alpha-IFN therapy and administration of diclofenac and indomethacin were discontinued and prednisolone therapy (50 mg daily) was begun. Hemoglobinuria disappeared by 5 days and laboratory data became in normal range by 3 weeks. Drug induced immune hemolytic anemia was diagnosed by the process of laboratory data, especially of direct Coombs' test. Though drug-induced immune hemolytic anemia is a rare adverse side effect during alpha-INF therapy for renal cell carcinoma, it seemed an important complication. PMID- 8709451 TI - More on critical thinking, clinical judgment, and documentation. PMID- 8709452 TI - Dispensing medications dangerous: physicians' support appreciated. PMID- 8709453 TI - ED nurse practitioner protocols and training sought by British nurse. PMID- 8709454 TI - Christian Science and emergency care: a case of reconciling conflicting beliefs. PMID- 8709455 TI - Pneumonic plague in a 15-year-old Utah girl. PMID- 8709456 TI - Health care beliefs and practices among the Pennsylvania Amish. PMID- 8709457 TI - Caring for Cambodian refugees in the emergency department. PMID- 8709458 TI - Caring for the Latter-day Saint patient. PMID- 8709459 TI - Surviving culture shock: lessons learned as a medical missionary in Jamaica. PMID- 8709460 TI - Caring for the Islamic patient. PMID- 8709461 TI - A look back: migraines. PMID- 8709462 TI - Interpreting needs of the ED patient: one California hospital's 3-week study. PMID- 8709463 TI - The Catholic patient in the emergency department. PMID- 8709464 TI - Thoughts on labels from a Chicana emergency nurse. PMID- 8709465 TI - ED culture, if not equipment, similar in Antigua. PMID- 8709466 TI - Thoughts on caring for transsexual patients. PMID- 8709467 TI - Developing and evaluating an emergency nursing orientation pathway. AB - OBJECTIVE: Orientee evaluation forms frequently focus on tasks and technical skills rather than decision-making skills. An orientation pathway was developed to evaluate orientee clinical decision-making skills. The pathway can assist in identifying orientees who require additional educational resources or interventions. Early interventions may decrease the length of orientation, increase an employee's length of stay in the institution, and contribute to cost savings. METHODS: The pathway was developed with employee exit interview data, preceptor interviews, identification of critical indicators, and the orientation literature. The pathway consists of six categories. Indicators within each category were identified and ranked according to four levels of proficiency (level 1 for the least proficient and level 4 for the most proficient). A convenience sample of seven ED orientee and preceptor pairs agreed to participate in the pilot use of the pathway. An expected timeline of progress had been developed by a group of 20 preceptors during a preceptor workshop. An average proficiency level (score) was obtained for each category for 12 weeks with the weekly scores plotted on the pathway. This level was compared with the expected proficiency level in each category for each week. Demographic information was obtained from the orientee and preceptor pairs to determine whether certain characteristics facilitate pathway progression. Data were analyzed with frequency distributions, and measures of central tendency and variance. Spearman's rank correlations were used to examine the proficiency level in each category for each week in relation to demographic characteristics of the preceptor and orientee. RESULTS: Orientees progressed faster than expected in all categories. It took 8 weeks for an orientee to attain the highest proficiency level in all categories. The fastest progress was made in the category " Accurately Evaluates Patient Responses." Slowest progress was made in the category "Safety in Blood and Drug Administration." Preceptors and orientees rated in the pathway as very useful. DISCUSSION: Further studies with the orientation pathway may help to determine the ideal time frame for orientation, characteristics of orientees who progress as expected or faster, and educational strategies to facilitate orientee progression and clinical decision making. PMID- 8709468 TI - Hemophilia emergencies. PMID- 8709469 TI - Increased use of unlicensed assistive personnel: pros and cons. PMID- 8709470 TI - The Edward Hospital Care Center. PMID- 8709471 TI - A pediatric care and resuscitation cart: one community hospital's ED experience. PMID- 8709472 TI - Race, culture, and medications. AB - Cultural, ethnic, and environmental influences on medical therapy add to the complexities encountered by practicing health care professionals. Ignoring such studies may inadvertently result in increased patient suffering, an increase in side effects, or both. Pharmacokinetic drug differences, especially polymorphism, are more relevant if the patient's condition is severely compromised and the drug has a narrow therapeutic range. However, if the drugs is carefully selected and the dosage is calculated according to the patient's therapeutic response, adverse outcomes can usually be reduced. PMID- 8709473 TI - Out-of-hospital thrombolytic administration. PMID- 8709474 TI - Justifying the cost of your air medical transport program. PMID- 8709475 TI - The importance of forensic photography in the emergency department. PMID- 8709476 TI - Cardiopulmonary arrest record. PMID- 8709477 TI - Special considerations for Orthodox Jewish patients in the emergency department. AB - Orthodox Jews are a special cultural group; their entire lives revolve around the teaching of the Torah. Their religious beliefs are reflected in all aspects of their lives, in both health and illness. The emergency department at Maimonides Medical Center has strived to serve this community and has modified common patient care practices to meet the needs of this special population. PMID- 8709478 TI - Computerized charting by exception at triage. PMID- 8709479 TI - Marketing the emergency department at Northern Nevada Medical Center--a nursing approach. PMID- 8709480 TI - Chest pain and syncope in a 15-year-old trauma patient. PMID- 8709481 TI - Foley catheter balloon technique for visualizing the hymen in female adolescent sexual abuse victims. PMID- 8709482 TI - Free medical care: Shriners hospitals offer that and more. PMID- 8709483 TI - Acute low back pain in an elderly woman. PMID- 8709484 TI - Deb Heiberger: advocate for the homeless. Interview by Marlene Jezierski. PMID- 8709485 TI - Belief. PMID- 8709487 TI - [Medical topics: polysurgical syndrome]. PMID- 8709488 TI - [SCOPE--reading the human genome]. PMID- 8709486 TI - I Corinthians 12:14. PMID- 8709489 TI - [Socioeconomic analysis of nursing services--changes in demand for nursing personnel]. PMID- 8709490 TI - [Specialist nurses in private practice--maintaining a private practice is a hard yet rewarding job]. PMID- 8709491 TI - [He knew the moment his son died thousands of miles away]. PMID- 8709492 TI - [Liberation of medical service--gifts or rewards from patients]. PMID- 8709493 TI - [Life of elderly patients at Farnham Hospital in England]. PMID- 8709494 TI - [New medical fee system and nursing service--terminal care of elderly people]. PMID- 8709495 TI - [Government plan to support child rearing in the age of fewer children]. PMID- 8709496 TI - [Health care and social services provided for the elderly in Hawaii: a study of the activities of Alzheimer's Disease and Related Diseases Association in Honolulu]. AB - The purpose of this study is to develop helpful strategies to serve the elderly and their families in Japan. We conducted the study by spending three months at the Honolulu Chapter of the Alzheimer's Association, participating in various programs of the Association, conducting a literature search and interviewing various health and social services available to the caregivers and persons with Alzheimer's disease in Hawaii. From the study we make the following recommendations, which we feel would be very helpful if applied to the Japanese context; 1) The supply of information through educational videos and leaflets in caregiver support center and so on. 2) Active efforts to make widely known the social services in public place. 3) The promotion and development of the voluntary activities to increase services and resources for Japanese. 4) The promotion of caregiver support groups in the health care system of each Japanese community. 5) QOL (Quality of Life) programs for dementia patients among elderly such as reminiscence and life review therapy. 6) The development of the organization with the help of professionals and scholars of related fields. PMID- 8709498 TI - [Activities of volunteers in hospitals. 20 years of hospital volunteer activities]. PMID- 8709497 TI - [Nursing solidarity to care for the Kobe earthquake victims]. PMID- 8709499 TI - [Activities of volunteers in hospitals. Volunteers are a great help for the nursing department]. PMID- 8709500 TI - [Activities of volunteers in hospitals. Volunteer activities in an independent hospice "Peace House"]. PMID- 8709501 TI - [Activities of volunteers in hospitals. Church members work as volunteers for library service and for sick children]. PMID- 8709502 TI - [Activities of volunteers in hospitals. Sick children are looking forward to visit of volunteers]. PMID- 8709503 TI - [Activities of volunteers in hospitals. Akebono Breast Cancer Support Service (ABCSS)]. PMID- 8709504 TI - [JNA nursing volunteers caring for the Kobe earthquake victims]. PMID- 8709505 TI - [How people were coping with the tragedy three weeks after the earthquake]. PMID- 8709506 TI - [More human resources planned and more financial resources explored for care of the elderly]. PMID- 8709507 TI - [Report of a hospital director of nursing--how to make a good director of nursing?]. PMID- 8709508 TI - [History of Japanese nursing that I lived: emergency nursing during the past major earthquakes]. PMID- 8709509 TI - [Report of a hospital director of nursing--network of hospital directors of nursing]. PMID- 8709510 TI - [Kobe earthquake, nurse placement centers on line, group home for senile elderly people]. PMID- 8709511 TI - [History of Japanese nursing that I lived: early nursing schools and nurses who studied in the United States]. PMID- 8709512 TI - [Medical topics: healing effect of holding hands over patients]. PMID- 8709513 TI - [Socioeconomic analysis of nursing service--salary structure for nursing personnel]. PMID- 8709514 TI - [Specialist nurses in private practice--home infusion therapy]. PMID- 8709515 TI - [Tradition of nursing in Europe]. PMID- 8709517 TI - [New medical fee system and nursing service--hospitalization management fee and care mix]. PMID- 8709516 TI - [Life of elderly patients at Farnham Hospital in England]. PMID- 8709518 TI - [Kobe earthquake follow up. Nursing volunteer activities in the disaseter stricken area]. PMID- 8709519 TI - [Kobe earthquake follow up. Letters from nurses working in the Kobe area]. PMID- 8709520 TI - [My first paper presentation at ICN Congress in Madrid]. PMID- 8709521 TI - [Controversy on state license of "psychiatric social worker" as a new category of health worker]. PMID- 8709522 TI - [Nursing care for foreigners in Japan. Need for understanding problems of alien patients in Japan]. PMID- 8709523 TI - [Nursing care for foreigners in Japan. Foreigner-friendly hospitals and health centers in Japan]. PMID- 8709524 TI - [Nursing care for foreigners in Japan. Health care experienced by foreign families living in Japan]. PMID- 8709525 TI - [Nursing care for foreigners in Japan. Population and maternal health statistics of aliens in Japan]. PMID- 8709526 TI - [Nursing care for foreigners in Japan. How do you say it to patients at the clinic in English, Chinese, Korean, Tagalog, or Thai?]. PMID- 8709527 TI - [Kobe earthquake follow ups. Nursing in hospitals and makeshift evacuation centers]. PMID- 8709528 TI - [Need for certified ET nurse]. PMID- 8709529 TI - [Who PHC collaborative center for nursing and midwifery]. PMID- 8709530 TI - [National health policy administrators conference, hospital function evaluation system, enhancing the community health program]. PMID- 8709531 TI - [Report of a hospital director of nursing--supported by nurse administrators network]. PMID- 8709532 TI - [History of Japanese nursing that I lived: Ms. Masu Yumaki and nursing of her time]. PMID- 8709533 TI - [Medical topics: importance of scrub]. PMID- 8709534 TI - [Specialist nurses in private practice--infusion center by nurses]. PMID- 8709535 TI - [Liberation of medical service--information about medical institutions not readily available]. PMID- 8709536 TI - [Life of elderly patients at Farnham Hospital in England]. PMID- 8709537 TI - [New medical fee system and nursing service--psychiatric nursing service]. PMID- 8709538 TI - [Nurses' knowledge of laboratory tests essential for collaborative nurse physician relations]. PMID- 8709539 TI - [Creation of clinical nurse specialist for ICU]. PMID- 8709541 TI - [Tragedy of fathers killing their sons]. PMID- 8709542 TI - [Efforts to eliminate private attendance in hospitals. Purpose of and need for elimination of private attendance]. PMID- 8709540 TI - [JNA position on the care system of the elderly and the proposed care insurance system]. PMID- 8709543 TI - [Efforts to eliminate private attendance in hospitals. An opportunity to realize nursing profession's self-reform]. PMID- 8709544 TI - [Efforts to eliminate private attendance in hospitals. Care mix helps elimination of private attendance in hospital]. PMID- 8709545 TI - [Efforts to eliminate private attendance in hospitals. Nursing service without private attendance successfully realized through care mix]. PMID- 8709547 TI - [Efforts to eliminate private attendance in hospitals. Transformation to quality nursing care]. PMID- 8709546 TI - [Efforts to eliminate private attendance in hospitals. Increase of staffing level essential]. PMID- 8709548 TI - [Efforts to eliminate private attendance in hospitals. My advice to eliminate private attendance successfully]. PMID- 8709549 TI - [Efforts to eliminate private attendance in hospitals. Efforts made by prefectural nursing associations]. PMID- 8709550 TI - [JNA proposal for a new care system toward the 21st century]. PMID- 8709551 TI - HTLV-I infections in offspring derived from HTLV-I carrier rats during the suckling period. AB - An experimental rat model for vertical transmission of human T cell leukemia virus type I (HTLV-I) was used to examine whether the infection of offspring derived from HTLV-I carrier rats could be established during the suckling period. MT-2 (2 x 10(7)) cells were injected into 5-week-old rats twice, at 2-week intervals. HTLV-I infected or non-infected female rats were mated with HTLV-I carrier male rats. The titer of serum antibodies against HTLV-I in the offspring derived from non-infected dams was less than 1:16 by the agglutination test during the suckling period. The serum antibodies of the offspring derived from the infected dams was less than 1:32 at 1 day after birth and increased steadily to 1:2048 at 14 days. However, the HTLV-I proviral sequences were not detected in any organs of the offspring during the suckling period as determined by the nested double polymerase chain reaction (PCR). These findings indicate that maternal antibodies against HTLV-I were not readily transmitted through the placenta and that the anti-HTLV-I antibodies in the offspring came from the milk of the dams. Furthermore, the HTLV-I infection in the offspring that was derived from the carrier dam may not be established during the suckling period but after weaning. PMID- 8709552 TI - A case report of rectal schwannoma. AB - We experienced a case of rectal schwannoma which was diagnosed before surgery. In this paper we have described this rare case and have discussed a number of similar cases reported in the literature. PMID- 8709553 TI - Automatic analysis of sleep spindles--assessment in one case treated with a benzodiazepine anxiolytic drug. AB - Wave-form analysis and detection of sleep spindles should be automatically performed to achieve objective and highly precise determination of the pattern of appearance of sleep spindles. The conditions for constructing the automatic analysis and detection are as follows: 1) it is recommended that EEGs be sampled at 500 Hz or higher intervals; 2) the digital filter with the threshold that is most appropriate for removal of artifacts should be used; 3) various conditions for the analysis and detection of wave-forms are widely used; and 4) a procedure for inspecting and reading record paper should also be included in the set-up. Taking these points into consideration, the authors developed an automatic analysis system for sleep spindles, which employs a computer. The system was applied to a case preliminarily treated with a benzodiazepine anxiolytic drug, ethyl loflazepate (CM6912). The assessment using this system revealed that the CM6912 administration induced an increase in sleep spindles, delayed frequency, and caused amplitude increases and a prolonged duration. PMID- 8709554 TI - A revised cold water immersion test for assessing peripheral circulatory function. AB - To improve the conventional cold water immersion test for assessing peripheral circulatory function, the immersed area of the hand was changed in this study. Twenty-seven healthy medical students with a mean age of 24 years participated in the study. The cold water immersion tests were carried out using two methods (in 5 degree C water for 1 min), in which the immersed area was up to the metacarpophalangeal (MP) joints (hereafter, MP-method) which is a new method, or to the wrist (hereafter, W-method) which is a conventional method. The recovery of skin temperature, vibratory perception threshold and complaints of finger pain were determined during cold immersion tests with the two methods. The mean skin temperature for the MP-method after immersion recovered more quickly than that for the W-method. However, the time courses of recovering were parallel for the two methods. Significant differences were noted at 3 min, 5 min and 10 min after cessation of immersion. There was a significant positive correlation in the recovery rates at 5 min after immersion between the two methods (correlation coefficient = 0.65, p < 0.01). The mean vibratory thresholds for the W-method were higher than those for the MP-method, and the difference was significant (p < 0.01). All subjects preferred the MP-method to the W-method because there was less finger pain. These results indicate that the MP-method, immersion up to the MP joint of a hand, for assessing peripheral circulatory function is better than the W-method. PMID- 8709555 TI - Reassessment of second look operation for epithelial ovarian cancer. AB - We retrospectively reviewed 91 patients with epithelial ovarian cancer who underwent second-look operation (SLO). SLO identified a persistent tumor in 18 (19.8%) of 91 patients. Of these 18 patients, 13 (72.2%) showed disease progression after SLO. Patients with a positive SLO in which the identification of the persistent disease was only possible by washing cytological study showed better prognosis than patients with macroscopic lesions. Of the 73 patients with a negative SLO, 12 (16.4%) developed recurrence after cessation of treatment. These 12 patients were from stage 1c to 3 with Grade 2 and 3 tumors. The SLO findings were more sensitive than the serum tumor marker in identifying persistent disease. Also we reviewed 55 patients having no SLO. Among 57 cases in stage 1 and 2 with Grade 1 tumor, there were no cases of recurrence, regardless of whether they had SLO or not. Our results suggest that SLO findings correlate well with the patient's prognosis. SLO may not be needed for patients in stage 1 and 2 with Grade 1 tumor. Patients with advanced stage disease with Grade 2 and 3 tumors have high-risk for recurrence after negative SLO. While SLO is still of benefit in management of ovarian cancer, refinements are needed to determine its indication and scheduling. PMID- 8709556 TI - Monoclonal antibodies (NU-T1 and NU-T2) recognizing antigens expressed on human T cells. AB - Two murine monoclonal antibodies (mAb) designated as NU-T1 and NU-T2 were generated to identify the novel antigens expressed on human T cells. NU-T1 reacted with peripheral blood T cells, but not with granulocytes or monocytes, whereas NU-T2 were not reactive with peripheral blood lymphocytes, granulocytes or monocytes. Both NU-T1 and NU-T2 reacted with thymocytes. The incidence of NU T1 or NU-T2 reactivity with the leukemic cells from patients with various types of leukemia was as follows: 12/12 or 4/12 in acute lymphocytic leukemia (ALL) of T cell type, 6/6 or 0/7 in adult T cell leukemia, 0/6 or 0/19 in ALL of non-T non B cell type, and 0/8 or 0/8 in acute myelocytic leukemia, respectively. The incidence of NU-T1 and NU-T2 reactivity with human leukemic T cell lines were 4/10 and 5/10, respectively. In contrast, neither NU-T1 nor NU-T2 reacted with B cell lines, non-T non-B cell lines, or myeloid cell lines tested. These results indicate that NU-T1 and NU-T2 recognize two distinct differentiation antigens expressed on the cells in the T cell lineage. These mAb would be useful reagents to study the function of human T cells and the immunophenotyping of human T cell malignancies. PMID- 8709557 TI - Experimental intraperitoneal administration of cisplatin-activated carbon for rat ovarian adenocarcinomas. AB - Selective delivery of a high level of cisplatin (CDDP) to the omentum, intraperitoneal disseminated tumors and lymph nodes was studied using a dosage form of CDDP-activated carbon. Although intraperitoneal CDDP-activated carbon delivered much higher levels of CDDP to the omentum, tumors and lymph nodes, a primary burst phenomenon of CDDP occurred just after administration. This indicates that activated carbon is an unsuitable particle to provide a slow steady release of CDDP. PMID- 8709558 TI - Hepatic arterial chemotherapy for liver metastases from colorectal cancer. AB - We have investigated the complications, toxicities, tumor response and survival rate with hepatic arterial (HA) chemotherapy for liver metastases from colorectal cancer. Forty-three patients with measurable liver metastases and 10 patients after the resection of liver metastases were treated with HA bolus mitomycin C (MMC) or continuous fluorouracil (5-FU) infusion between 1986 and 1994. The catheter- or reservoir-related complications such as catheter induced infection, subcutaneous reservoir pocket infection, or catheter or hepatic artery occlusion occurred in 14 patients (26%) mainly in our early cases. Severe gastritis or a gastroduodenal ulcer developed in 12 patients (23%), in particular after treatment with continuous infusion of 5-FU and when the catheter was placed into the gastroduodenal artery during laparotomy. An ulcer often caused a serious complication such as a duodeno-biliary fistula, penetration into the hepatic artery or duodenal perforation. Also noted were upper gastrointestinal symptoms in 26 patients, hepatic toxicities in 3 patients, and bone marrow depression in 4 patients. Because of these complications and toxicities, HA therapy was discontinued in more than 60% of the cases. The overall tumor response rate was 17%. However, in the cases which were treated by 5-FU continuous infusion for longer than 3 months, the response rate was higher than 60%. There was no significant difference in survival rate between HA therapy and systemic palliative chemotherapy. These results suggest that it is important to prevent gastrointestinal toxicities in order to allow continuation of HA continuous 5-FU infusion therapy which could provide a higher response rate and a better survival for colorectal cancer patients with liver metastases. PMID- 8709559 TI - Health care of villagers in northeast Thailand--a health diary study. AB - A one-month health diary was used as a research instrument for measuring health care in the northeastern rural area of Thailand during the month of November, 1992. Three hundred forty-five respondents, which represents 22.2% of the total households in 12 villages from 2 districts in the Khon Kaen province of northeast Thailand, completed the health diaries. Self-medication was most practiced for health care (37.5% of ill persons) followed by health service utilization at local health center (19.4%), at community hospital (11.4%), at private clinic (11.4%), and by "wait and see" (9.0%). Analgesics were the most frequently used drugs for relief of fever, headache, common cold and abdominal pain. The use of a combination of drug (eg. a cocktail) was common as self-medication for relief of back and leg pain. Traditional or herbal drugs were a frequently chosen alternative for self-medication, both in adult and children. The authors suggest that the villagers should be educated concerning drug hazards in order to increase their background knowledge on drug use. PMID- 8709560 TI - Expression of MAGE gene family in lung cancers. AB - MAGE gene family encodes tumor-specific antigens recognized on HLA-A1 by cytotoxic T lymphocytes. We have investigated the mRNA expression of MAGE gene family in lung cancers using MAGE gene specific primers, reverse transcription polymerase chain reaction (RT-PCR) and restriction enzyme analysis. MAGE-1, -3, 4, -41 or -6 gene was respectively expressed in 6, 5, 6, 7, and 4 of 53 lung cancer tissues (8 to 13%) or in 0, 6, 6, 7 or 8 of 14 lung cancer cell lines (0 to 53%). These results suggest that a substantial proportion of lung cancers express MAGE genes. PMID- 8709561 TI - Diffuse panbronchiolitis--a thin-section CT scoring system. AB - The usefulness of thin-section computed tomography (CT) in the identification of the specific and clinically relevant pathologic changes in airway disease has been well documented. However, this approach has not been used for objective evaluation in patients with diffuse panbronchiolitis. Therefore attempts were made to use it in patients with diffuse panbronchiolitis. The study group included 12 men and 10 women, 17 to 69 years of age (mean, 47 years) with diffuse panbronchiolitis. All patients were evaluated by thin-section CT and pulmonary function testing. The CT score was calculated by a modified score criteria. The correlations between the CT score and the pulmonary function parameters were assessed by Pearson's linear regression analysis. Bronchiolar inflammation and airway-ectasia were present in all 22 patients. Periairway thickening was present in 17 patients. Pus plugging was present in 18 patients. Two patients had bullae formation, and 6 had air-trapping. Pulmonary collapse or consolidation was evident in 6 patients. The CT score demonstrated a significant negative correlation with FEV1% (r = -3.267, p = .0230), %VC (r = -4.658, p = .0018), and arterial blood oxygen concentration (r = -1.009, p = .0381). The thin-section CT findings in patients with diffuse panbronchiolitis closely resembled those in patients with cystic fibrosis, previously described. Furthermore, a CT scoring system is useful for objective evaluation of lung disease severity in patients with diffuse panbronchiolitis. PMID- 8709562 TI - A patient with chronic hepatitis C and a history of abuse of analeptic drugs, who showed hallucination and delusion with interferon administration. AB - We experienced a case of chronic hepatitis type C accompanied with hallucination and delusion induced by interferon (IFN) therapy positive. The case was a 47-year old male, whose laboratory data showed positive for anti-Hepatitis C Virus (HCV) and elevated transaminase level. He was treated with 6 MU/day of natural-type IFN alpha (HLBI). Sleeplessness and delusions of persecution developed about 2 months after the start of IFN therapy. The interview of the psychiatrist disclosed that the patient had a history of addiction to drugs, and these psychiatric symptoms were diagnosed as being of "the flashback phenomenon." These side effects were improved after the administration of psychotropics and it was suggested that we had to take care of the development of flashback phenomenon during the treatment of IFN in cases of chronic hepatitis with a history of addiction to drugs. PMID- 8709563 TI - A case of obstructive icterus caused by incarceration of a pancreatic stone in the common channel of the pancreatobiliary ducts. AB - A 51-year-old man had repeated admissions and discharges from the hospital for alcohol dependence and alcoholic hepatitis. Thereafter, with a diagnosis of chronic pancreatitis, he was examined on an outpatient basis. He presented with right hypochondriac pain as the chief complaint, and was admitted to the hospital because of a suspected acute exacerbation of chronic pancreatitis. Pyrexia and jaundice were present from about the 5th hospital day, and he was admitted to Kurume University Hospital with a diagnosis of obstructive icterus. Percutaneous transhepatic biliary tract drainage (PTBD) was immediately performed. A roentgenogram revealed moderate dilatation of the extrahepatic bile ducts and stricture of the intrapancreatic common bile duct. No calculi were observed in the gallbladder or bile ducts by ultrasonography of the abdominal region. The main pancreatic duct was dilated to 10 mm, and a strong echo, probably a calculus, was observed in the main pancreatic duct. Pancreatoduodenectomy was performed with a diagnosis of stricture of the common bile duct due to chronic pancreatitis with a pancreatic stone. Examination of the excised specimens revealed a rigid pancreatic head; and when the common bile duct was incised, no stricture was observed. A milk-white calculus about 10 mm in diameter was observed in the common channel of the lower portion of the common bile duct. The orifice of the pancreatic duct was also observed at the same site. The common channel was about 20 mm long, and malfusion of the pancreatobiliary ducts was observed. The obstructive icterus was due to an impaction of the pancreatic stone at the confluence of the pancreatobiliary ducts. The calculus was composed of calcium carbonate, and the histopathological findings in the pancreas showed chronic pancreatitis. PMID- 8709564 TI - Recurrent herpetic infection of the oral floor--a case report. AB - Recurrent herpetic infection on the mobile mucosa such as the oral floor, has not been reported previously. A case of recurrent herpetic infection of the oral floor in a 52-year-old man is presented. This case illustrates the fact that recurrent herpetic lesions can occur anywhere in the intra-oral mucosa. PMID- 8709565 TI - Superficial spreading-type mucosa-associated lymphoid tissue (MALT) lymphoma in the stomach--a case report. AB - We presented a case of a 71-year-old woman who presented irregular mucosal folds with tiny depressions and multiple erosions with ulceration in the stomach, on endoscopy and roentgenography. Histological findings from biopsy specimens showed dense and mildly atypical lymphoid-like cells invading the submucosa. Immunohistochemical examinations revealed monoclonal reactivity of Heavy-chain. She was then diagnosed to have low grade malignant lymphoma in the stomach and underwent total gastrectomy. The resected cancer demonstrated the pathological features including small lymphocytic plasmacytoid cells and lymphoepithelial lesions characteristic of a mucosa-associated lymphoid tissue (MALT) lymphoma. The lymphoma cells expressed B cell markers with a phenotype of IgG and lambda type. It is difficult to make a firm diagnosis of a MALT lymphoma from only small biopsy specimens. Accordingly it is necessary to complete the observed clinical process, endoscopic and roentgenographic findings with immunohistochemical characteristics to determine diagnosis. The MALT lymphoma should be treated as a low-grade malignancy and diagnosis sufficiently early can lead to a favorable prognosis. PMID- 8709566 TI - Oral cancer and hepatitis C virus (HCV): can HCV alone cause oral cancer?--a case report. AB - Previously, we reported the high prevalence of hepatitis C virus (HCV) infection in patients with oral cancer or oral lichen planus in Kyushu, Japan. We now report a 61-year-old man with chronic hepatitis C and no oral lesions who developed oral cancer 6 months after interferon therapy (interferon alpha [HLBI], 6 million units (MU) daily for 2 weeks and then 3 times a week for 14 weeks). This case emphasizes the need for periodic oral cavity examinations of hepatitis C patients and contributed to the investigation of oral cancer and HCV. PMID- 8709567 TI - The use of a novel operant test to determine the strength of preference for flooring in laboratory rats. AB - A previous study showed that laboratory rats preferred to dwell on a solid floor rather than a grid one, particularly when resting (Manser et al. 1995). The strengths of this preference was investigated in an operant trial using a novel test apparatus, which consisted of a grid-floored cage and a solid-floored cage, joined via a central box containing a barrier whose weight was adjustable. Trials in which rats had to lift the barrier in order to explore the whole apparatus were alternated with those in which the rats were confined on the grid floor and then had to lift the barrier in order to reach the solid floor. The latter trials were carried out at the beginning of the light period when the rats were seeking a resting place. In both trials, the weight of the barrier was progressively increased for each rat, until a maximum weight was found which it would lift either to explore its environment (weight A) or to reach the solid floor (weight B). No significant differences were found between weights A and B, showing that rats would work as hard to reach a solid floor to rest on as they would to explore a novel environment. The apparatus used could with some modifications, be appropriate for use in other operant studies in laboratory rats. PMID- 8709568 TI - Behaviour of rats in their home cages: daytime variations and effects of routine husbandry procedures analysed by time sampling techniques. AB - The behaviour of laboratory rats in their home cages was observed on both the mornings and the afternoons of days when cages were cleaned and compared to days when cages were not cleaned. Two different time sampling methods, 'instantaneous sampling' and 'one/zero sampling', were used and compared. In general the rats were more active in the mornings than in the afternoons. Activity, particularly locomotion and that associated with manipulation of the bedding was increased during both the mornings and the afternoons of cleaning days. Defaecation also increased on cleaning days whereas sitting decreased. The cleaning regime appeared to have a greater effect on behaviour than did time of day and the effect of cleaning lasted for several hours after the procedure had been completed. The implications for experimental design are discussed. PMID- 8709569 TI - The influence of body weight on open field and swimming maze performance during the post-weaning period in the rat. AB - The open field and swimming maze tests are frequently used as part of a behavioural assessment battery in reproduction or developmental toxicology studies in the rat. Interpretation of the results of these tests is very difficult when the body weight of the rats is not similar in the various treatment groups. In order to better understand this problem, data from approximately 500 untreated rats tested under identical conditions were analysed with respect to body weight. No correlation was found in water maze performance or open field motor activity. A very slight but significant correlation was identified, however, in the time spent by female rats in various regions of the open field, with the smaller rats spending more time in the corners of the testing arena. PMID- 8709570 TI - Lung, ileum and heart are predilection sites for AApoAII amyloid deposition in CD 1 Swiss mice used for toxicity studies. Pulmonary amyloid indicates AApoAII. AB - Amyloid deposits represent frequent histological findings in SPF strains of mice mainly used for toxicological studies. Usually, these are deposits of reactive amyloid (AA-amyloid) derived from the acute phase protein serum amyloid A (SAA). The SAA is an apoprotein of high density lipoprotein (apoSAA). Senescence accelerated amyloid (ASsam) occurs in a special strain of mice. This type of amyloid is derived from apolipoprotein-AII and, therefore, is called AApoAII. Recently, C57B1/Ka control mice not treated for long duration with immunosuppressive agents, were found to have developed AApoAII-amyloidosis with a predilection for the deposits in the ileum (HogenEsch et al. 1993). In the present study, SPF CD-1 Swiss outbred mice, used for chronic toxicity experiments, were investigated. Amyloidosis was diagnosed by haematoxylin and eosin staining. The tissue localization of amyloid was recorded and confirmed by Congo red staining. The chemical type of amyloid was investigated by peroxidase antiperoxidase (PAP)-immunostaining using anti-murine AA and anti-murine AApoAII antibodies. Those animals which died during the study and the mice killed at end of the experiment, aged 18 months, from treated as well as non-treated control groups, showed AApoAII-amyloid deposits with similar prevalence. The AApoAII amyloid had organ predilection for gut, heart and lung tissue. A group of animals was euthanazed intercurrently at a young age, since they suffered from spontaneous dermatitis associated with Staphylococcus aureus infection. Sixty eight percent had reactive amyloid deposits found primarily in spleen, liver, kidney and gut. From these findings and literature data on various other mouse strains, it is concluded that in mice used for toxicity studies, AA and AApoAII types of amyloidosis may be expected. The deposition patterns of these types of amyloid are slightly different. AA-amyloid has a predilection for spleen, liver, gut and kidney, and is often associated with inflammatory lesions of the skin, whereas masses of amyloid in lung, heart and ileum suggest AApoAII. Pulmonary amyloid appears to represent the most reliable deposition criterion for discriminating between both types of amyloidosis. PMID- 8709571 TI - Epi- and metaphyseal morphology in the long bones of BDIX/Han rats. AB - Development and morphology of the epiphyses of the long bones were investigated in 93 adult rats of 7 different strains (BDIX/Han, BDE/Han, BN/Han, DA/Han, LEW/Han, AVN/IpcV/Wistar/Rehbrucke, Shoe: WIST) from the age of 14 weeks up to the age of 78 weeks. Strain-related differences were found in the development of the secondary centre of ossification, which was retarded in the BDIX/Han rats. Furthermore, closure of the growth plate started earlier in the BDIX/Han rats. In addition various regressive changes were detected in the growth plates of long bones of all rats, but not of the ribs. The frequency and extent of these changes varied between individuals and strains. Degeneration of the matrix and necrosis were already observed at 14 weeks of age. PMID- 8709572 TI - Mutual viral and bacterial infections after housing rats of various breeders within an experimental unit. AB - Fifteen athymic rat strains from 11 breeding colonies were housed within an experimental facility for an immunological study. Health status records supplied with 14 of the strains listed infections by Kilham's rat virus (KRV), Clostridium piliforme (Bacillus piliformis) and Pasteurella pneumotropica for 2, 2 and 1 colonies respectively. In sera taken previous to the study from euthymic rats of 10 strains, antibodies to KRV were detected in 3 strains, to Pneumonia virus of mice (PVM), Rat corona virus (RCV) and Sendai virus in one strain each and to P. pneumotropica in 2 strains. Only 2 of the KRV infections had been reported by the supplier. During the study rats of all 10 strains developed antibodies to 2-4 of viral antigens. Eight out of 10 rat strains seroconverted to 1-5 of the antigens C. piliforme (B. piliformis), Bordetella bronchiseptica, Haemophilus spp., P. pneumotropica and Streptobacillus moniliformis. Two rat strains housed in filtertop cages did not develop antibodies to bacterial antigens. The potential detrimental effects of intercurrent infections on the outcome of the comparative immunological study are discussed. PMID- 8709573 TI - Translocation of bacteria from the gastrointestinal tract in immunodeficient mice. AB - Host defence mechanisms associated with the inhibition of translocation of bacteria from the gastrointestinal (GI) tract were investigated in SCID and beige mice after decontamination with oral antibiotics and colonization with Escherichia coli C25. SCID mice, which have impaired T and B cell function, tended to have a greater incidence of bacterial translocation from the GI tract up to 7 days after inoculation compared with controls. However, after 7 days both SCID and controls cleared the E. coli C25 from the liver, spleen, blood and peritoneal cavity. Beige mice, with impaired NK cell and polymorphonuclear leukocyte function, were not able to clear the inoculated bacteria from their liver by 14 days after inoculation although the controls were cleared by 7 days. Numbers of bacteria in the mesenteric lymph nodes (MLN) of beige mice did not decrease significantly by 14 days after inoculation, whereas numbers in SCID mice decreased markedly within 7 days. These results suggest that defence mechanisms other than T and B cell function are important in the inhibition of systemic infection from the GI tract. PMID- 8709574 TI - Polymyopathy in a Syrian golden hamster. AB - A Syrian golden hamster suffered from general swelling of skeletal muscles. At microscopical observation the muscle tissue exhibited degeneration and necrosis, as well as regenerative features. The inflammatory response was very slight. The histopathological lesions were diagnosed as polymyopathy. PMID- 8709575 TI - Acid-base status and cardiovascular function in mink (Mustela vison) anaesthetized with ketamine/midazolam. AB - Heart rate, arterial blood pressure and blood acid-base status were determined in 18 adult female mink (mean (+/- SEM) body weight 1052 +/- 34 g) during long-term anaesthesia with either controlled ventilation (n=12) or spontaneous respiration (n=6). Surgical anaesthesia was induced by intramuscular injection of ketamine hydrochloride (Ketaminol Vet, 40.0 +/- 1.7 mg/kg) and midazolam hydrochloride (Dormicum 2.8 +/- 0.1 mg/kg) and maintained for at least 5 h by continuous intravenous infusion of this drug combination in 0.9% saline. For all animals, the mean rates of infusion of ketamine and midazolam were 48.4 +/- 1.6 and 1.61 +/- 0.12 mg/h, respectively. Following continuous infusion of the anaesthetics in isotonic saline, at a rate of 20 ml/h, a moderate 'dilution acidosis' developed, which could be corrected by replacement of part of the saline with sodium bicarbonate to a final concentration of approximately 25 mmol NaHCO3 per litre. However, when the animals were allowed to breathe spontaneously, an increase in heart rate and a combined respiratory and metabolic acidosis occurred, due to severe respiratory depression. Apart from these effects and a few cases of increased salivation, no adverse effects over time were observed on the arterial blood acid-base status and cardiovascular function of the animals during ketamine/midazolam anaesthesia. It is concluded that the procedure described for long-term anaesthesia in mink is convenient and safe for acute physiological experiments in this species, provided normal body temperature and pulmonary gas exchange is sufficiently maintained. Thus, the need for an adequately controlled artificial ventilation is strongly emphasized. Finally, a proposal for the composition of an intravenous solution, containing ketamine and midazolam hydrochloride, and sodium bicarbonate in saline, suitable for long-term anaesthesia in adult mink is presented. PMID- 8709576 TI - Induction of anaesthesia with halothane and isoflurane in the rabbit: a comparison of the use of a face-mask or an anaesthetic chamber. AB - The effects of induction of anaesthesia with halothane or isoflurane were studied in rabbits. The anaesthetic agents were delivered either via a face-mask, or the animals were placed in an anaesthetic induction chamber. All rabbits had periods of apnoea during induction, lasting 30-120 s, resulting in moderate hypercapnia and acidosis. Periods of apnoea were associated with a marked bradycardia. The combination of bradycardia and hypercapnia during induction may represent an increased risk of anaesthetic associated mortality. Animals in all groups tried to avoid inhaling anaesthetic vapour, and this behaviour, together with the occurrence of breath-holding suggests that induction was aversive. PMID- 8709577 TI - The telemetric monitoring of heart rate, locomotor activity, and body temperature in mice and voles (Microtus arvalis) during ambient temperature changes. AB - We have studied the physiological and behavioural responses in small rodents to ambient alterations. For this purpose, voles and mice were exposed to relatively low (12 degrees C) and high (35 degrees C) ambient temperatures, and heart rate (HR), locomotor activity (LA) and body temperature (BT) were recorded using telemetry system. The control HR (at 24 degrees C) of voles was lower than that of mice. The 'heat exposure' decreased HR to 85.0 +/- 3.3% in voles, and to 78.0 +/- 3.2% in mice compared with the mean HR of the same time in the control day. The 'cold exposure' increased the HR to 131.9 +/- 8.8% in voles, and 119 +/- 10.9% in mice. The decreasing rate of HR in heat exposure was smaller in voles than mice, and in cold exposure the increased rate was larger in voles than mice. Cold exposure decreased BT in both species; 96.1 +/- 0.5% in voles and 93.7 +/- 1.0% in mice. The LA was not changed significantly by heat exposure in either species, but was partially increased by cold exposure. These results demonstrate that telemetry was helpful for qualitative and quantitative behavioural studies in small rodents, and confirmed that the physiological and behavioural responses to ambient temperature changes differed between these animals. PMID- 8709578 TI - A new system for continuous intravenous infusion of pigs. AB - The swivel system described has been used by ILOB-TNO for prolonged periods of continuous infusion. It allows for less restricted movement of animals. The swivel prevents the infusion tube from twisting. The swivel system is simple, safe to operate and cost-effective. Animals have greater movement, can stretch and turn around. The physical discomfort and psychological stress of restricted movement are greatly reduced. The potentially complicating/confusing effects on the measurements taken during an experiment are reduced. PMID- 8709579 TI - The dust content, dust generation, ammonia production, and absorption properties of three different rodent bedding types. AB - Vermiculite, pine shavings and unbleached eucalyptus pulp contact bedding were compared for dust content, dust production (as a result of animal activity), moisture absorption properties and ammonia production. The percentage of dust particles <300 microm found in pine shavings and vermiculite was 18 and 8 times greater respectively than was found in eucalyptus pulp. The lowest ammonia levels (<1-1 ppm), measured on day 7, were obtained using eucalyptus pulp. Although the ammonia levels obtained for vermiculite compared very well with those of pulp, high levels of dust, up to 5 times more than pulp, were experienced with vermiculite. Pine shavings were associated with the highest ammonia concentration (1-3 ppm), but produced approximately 50% less dust than vermiculite. The water absorption properties obtained for pulp were between 35% and 76% higher than those obtained for vermiculite and pine shavings respectively. PMID- 8709580 TI - The cost-effectiveness of treatment for alcoholism: a second approximation. AB - OBJECTIVE: This review builds on the innovative research synthesis of Holder and his colleagues, addresses some of the limitations of the box-score approach to assessing treatment effectiveness that they used and provides a second approximation of the cost-effectiveness of treatment for alcoholism. METHOD: For each of 141 comparative treatment studies, we determined whether or not it found at least one statistically significant positive effect on a drinking-related outcome variable for each of the modalities examined in a paired contrast with one other condition. We next calculated the predicted probability of each study yielding at least one statistically significant treatment effect, based on the number of tests for treatment effects conducted. Following that, for each study of a particular treatment modality, the strength of the "weakest competitor" against which the modality had been compared was determined. For each modality, we used the average predicted probability of the relevant studies finding a significant effect and the average effectiveness of the weakest competitor to predict the modality's effectiveness. RESULTS: We calculated an Adjusted Effectiveness Index (AEIn) for each modality, which was the difference between its predicted and actual effectiveness score. Our AEIn results were consistent with those of Holder et al. in suggesting that some of the same modalities appear to be effective or ineffective. Our results differed from their findings with respect to other modalities, however. Using data presented by Holder and his colleagues on the minimum estimated cost of providing different modalities, we offer a second approximation of the modalities' cost-effectiveness. CONCLUSIONS: Overall, we found a smaller range of effectiveness across modalities than did Holder and his colleagues and a nonsignificant relationship between cost and effectiveness. Like Holder et al., we do not believe major treatment provision or funding decisions should be based solely on this type of review. PMID- 8709581 TI - Regional differences in alcohol and fatal injury: a comparison of data from two county coroners. AB - OBJECTIVE: While alcohol has been found to be associated with fatal injury, this association may vary in different regions of the United States where drinking patterns and problems are known to vary. The purpose of this study was to analyze alcohol in injury fatalities in a "dry" area of the country (Hinds County, Miss.) compared to a "wet" area (Contra Costa County, Calif.). METHOD: Data on all unnatural causes of death during a 1-year period among those 18 years and older were abstracted from county coroner records in Hinds County, Miss. (n = 222) and Contra Costa County, Calif. (n = 304). RESULTS: Across all causes of death, those in Hinds County were more likely to be alcohol-involved (57%) compared to those in Contra Costa (40%). Those in Hinds County were twice as likely as those in Contra Costa to have died from homicide (32% vs 17%), but the proportion of homicides which was alcohol positive did not differ between the two regions (51% vs 46%). Using logistic regression, gender (male) and region (South) were predictive of a positive blood alcohol concentration across all causes of fatality combined. Age (younger) and ethnicity (black) were predictive of homicide. Ethnicity (non-black), a negative blood alcohol concentration and a positive drug screen were predictive of suicide, while age (younger), ethnicity (non-black), a positive blood alcohol concentration and a negative drug screen were predictive of motor vehicle accidents. CONCLUSIONS: These data suggest that regional variations in alcohol's association with fatal injury do exist, and vary by demographic characteristics and cause of death. In addition, areas of the country that have relatively low rates of per capita consumption may have higher rates of alcohol-related injury fatalities than areas with greater per capita consumption. PMID- 8709582 TI - Alcoholism and typology: findings in an Irish private hospital population. AB - OBJECTIVE: To assess the validity of alcoholic typology in an Irish private hospital population. METHOD: We interviewed a randomly selected group of inpatient and outpatient alcoholics (N = 48, 42 male) in a private hospital in Ireland. Subjects had a structured psychiatric interview, including completion of a severity of alcohol dependence questionnaire, a Hamilton depression rating scale and a typology questionnaire. RESULTS: We found that there were differences between groups of alcoholics when divided according to Cloninger's typology hypothesis and by Von Knorring's age of onset criteria. The Type 2 early age of onset alcoholics (n = 23) had a significantly higher percentage of positive family histories of alcoholism, (p < .01), a higher percentage of sociopathic traits (p < .01) and increased severity of alcoholism (p < .05), relative to the Type 1 late age of onset alcoholics (n = 25). The Type 2 alcoholics also had increased daily consumption of alcohol in the month prior to admission, poor behavioral tolerance to the effects of alcohol and more fights that the Type 1 alcoholics. CONCLUSIONS: We concluded that there is validity to alcohol typology theory in relation to this Irish private hospital population and that age of onset is a useful defining criterion. PMID- 8709583 TI - Alcoholic beverage consumption and associated factors in Porto Alegre, a southern Brazilian city: a population-based survey. AB - OBJECTIVE: To describe the pattern of alcoholic beverage consumption and the prevalence of at risk drinking behaviors, as well as their association with demographic and socioeconomic factors in the adult population of Porto Alegre, a southern Brazilian city. METHOD: In a cross-sectional, population-based, multistage random sampling study, 1,091 (600 female) individuals (92% of those eligible) were selected and interviewed at home. Exposure to alcohol was measured by the CAGE questionnaire and by inquiring about the type, quantity and frequency of alcoholic beverage consumption. An average consumption of 30 g per day or more, a level of exposure associated with health risks, was considered as heavy drinking. Two positive answers to the GAGE questionnaire represented the cutoff for indicating dependence. RESULTS: The prevalences were: 9.3% (95% CI: 7.6 to 11.0) for dependence, 15.5% (13.4 to 17.7) for heavy drinking and 12.3% (10.4 to 14.2) for daily drinking; 24.1% (21.7 to 26.6) were abstinent. Women consumed alcoholic beverages in lower frequency and amounts than men. The most widely consumed beverages were beer, wine and "cachaca," a Brazilian sugarcane spirit. In a logistic regression model, increasing age, lower education and income, and nonwhite race were associated with heavy drinking and dependence. Households with 3-4 persons were associated with the lowest risk of heavy drinking, but the prevalence of dependence was higher in crowded households. The presence of another heavy drinker or dependent in the household was associated with heavy drinking but not with dependence. CONCLUSIONS: The study characterized a detailed pattern of alcoholic beverage use and indicated that at risk drinking is an important public health problem in a developing country. The risk factors for heavy drinking and dependence were the same, with the exception of age at starting to drink, heavy drinking or dependence-positive household members. PMID- 8709584 TI - An effect of alcohol on the distribution of spatial attention. AB - OBJECTIVE: Previous research on the effects of alcohol on visual performance led us to the prediction that alcohol should interfere with the distribution of visual spatial attention. This prediction was examined in two experiments that measured the effect of alcohol on reaction time (RT) for tasks that differed in visual spatial attention requirements. METHOD: In the first experiment, 48 adult volunteers (33 female) responded to either the onset or offset of one of five potential targets without alcohol to determine the relative demands on attention of stimulus onset and offset. The spatial extent of the five-target display was also varied. In the second experiment, the effect of alcohol was determined for both the onset and the offset tasks in 12 adult volunteers (nine female). RESULTS: The offset task was found to place greater demands on spatial attention as the increase in display area result in relatively greater increases in RT. Alcohol increased RT in the offset task for the larger, but not the smaller, display, and there were no significant effects of alcohol for the onset task. CONCLUSIONS: The results indicate that alcohol impairs performance on tasks that place greater demands on visual spatial attention and likely disrupts the ability to shift attention from one spatial locus to another during serial search. PMID- 8709585 TI - Investigator-observed alcohol-induced flushing but not self-report of flushing is a valid predictor of ALDH2 genotype. AB - OBJECTIVE: This article presents data on the validity of using self-report of alcohol-induced flushing and actual investigator-observed flushing following alcohol challenge to predict ALDH2 genotype in Asian-American men. METHOD: Men between the ages of 21 and 25 years who were of Chinese, Japanese or Korean descent completed questionnaires about their drinking history and their alcohol induced flushing history and associated symptoms. Fifty men selected for participation in the study were genotyped for alleles of ALDH2 and individually tested on two separate occasions following oral administration of placebo and 0.75 ml/kg (0.56 g/kg) alcohol. Facial flushing was assessed at baseline and at intervals over a 150-minute period after drinking using observational ratings. RESULTS: By comparing the results of ALDH2 genotype with investigator-observed flushing and with previous self-report of facial flushing, it was found that investigator-observed flushing is both a sensitive (100%) and specific (96%) predictor of ALDH2 genotype, whereas self-report of facial flushing is a sensitive (100%) but not a specific (68%) predictor of ALDH2 genotype. CONCLUSIONS: The results suggest that investigator-observed flushing provides a valid estimate of ALDH2 genotype, but that self-report of facial flushing is biased, giving a substantial overestimate. Due to an increase in error variance, studies that rely solely on self-report of flushing will more often lead to the conclusion that no association exists. This study supports the importance of using ALDH2 genotype, rather than self-report of flushing or ALDH2 phenotype, when examining factors associated with differences in drinking behavior, response to alcohol and risk for alcoholism or alcohol-related disease among Asians. PMID- 8709586 TI - Contradictory subject response in longitudinal research. AB - OBJECTIVE: This study investigated the prevalence of logical and estimation errors in a 4-year longitudinal survey of substance abuse for elementary and junior high school students. Logical errors occur when a subject reports having used a substance on one measurement occasion, but reports never having used the substance on a second measurement occasion. Estimation errors occur when a subject reports a lifetime frequency of use on a later survey that is less than the lifetime frequency of use reported for an earlier survey. METHOD: Substance abuse surveys were completed every year for four years by predominantly black District of Columbia students, beginning when the students were in 4th/5th grade, and ending when they were in 7th/8th grade. (Ns ranged from 5,671 to 6,421). Errors for alcohol, cigarette and marijuana use were calculated for every pair of years. Background characteristics such as demographic and academic performance variables were also assessed for each of the four years. RESULTS: Logical errors decreased for all substances as student got older, whereas estimation errors remained stable. Marijuana had the highest frequency of logical errors in every pair of years except one, and had the lowest frequency of estimation errors. Students who made errors in Years 1-2 were 1.64 times more likely to make errors again in Years 3-4. For some of the years, students who made any kind of error had lower school grades, were judged untrustworthy by their peers and were more likely to be males. CONCLUSIONS: The alarming prevalence of errors should be considered when conducting longitudinal substance abuse research. Several methods of dealing with these errors are reviewed. In most cases, the best a researcher can do is assess the error rates and report them. PMID- 8709587 TI - Alcohol outcome expectancies and alcohol use and problems. AB - OBJECTIVE: Numerous studies have demonstrated that alcohol outcome expectancies are strong correlates of various aspects of alcohol use. However, it has been suggested that forced-choice alcohol expectancy items may not measure the most salient anticipated effects of alcohol for an individual, and thus may create superficial responses. Additionally, research on attitude-behavior consistency has demonstrated that the nature of how an attitude is elicited can lead to attitudes that are more or less cognitive. The present study examined the applicability of this aspect of attitude theory to the study of alcohol outcome expectancies and the utility of self-generated outcome expectancies and subjective evaluations in predicting alcohol use and problems. METHOD: In an extension of methodology previously used in attitude research, alcohol-using college students (N = 165, 87 female) generated alcohol expectancies in response to two sentence stems designed to elicit "affectively" and "cognitively" based alcohol expectancies and made subjective evaluations (SE) ratings of each item generated on dimensions of positivity, likelihood and immediacy. RESULTS: While no support was found for the generalizability of a particular aspect of attitude theory, several interesting findings emerged. SE ratings were significantly related to measures of alcohol use and problems and, for alcohol use measures, predicted comparably to a standardized measure of expectancies. For alcohol dependence symptoms, the number of expectancies generated predicted significant unique variance over that accounted for by subjective evaluations. CONCLUSIONS: These results indicate that the sheer number of alcohol expectancies accessible to an individual may be an important correlate of alcohol dependence, independent of subjective evaluations. PMID- 8709588 TI - Getting drunk and growing up: trajectories of frequent binge drinking during the transition to young adulthood. AB - OBJECTIVE: The purpose of this study was: (1) to identify different trajectories of frequent binge drinking during the transition to young adulthood; (2) to validate the trajectories by relating them to behaviors and attitudes concerning alcohol and other drug use; and (3) to distinguish among the trajectories according to demographic characteristics and lifestyle experiences typical of the transition to young adulthood. METHOD: Four waves of national panel data were obtained from the Monitoring the Future project; 9,945 weighted cases from the 1976-85 high school senior year cohorts were surveyed at biennial intervals between ages 18 and 24. Frequent binge drinking was defined as having five or more drinks in a row at least twice in the past two weeks. RESULTS: Six distinct frequent binge drinking trajectory groups were specified a priori and confirmed with cluster analysis: Never, Rare, Chronic, Decreased, Increased and "Fling." Repeated measures ANOVAS revealed that the trajectories corresponded to patterns of change and stability in problems with alcohol, attitudes about heavy drinking, peer heavy drinking and illicit drug use. Results from logistic regression analyses predicting diverging and converging trajectories provided some support for the general hypothesis that trajectories of Chronic and Increased frequent binge drinking over time are associated with difficulties in negotiating the transition to young adulthood. CONCLUSIONS: The findings provide strong evidence for wide developmental variation in drinking patterns in the population, variation that is obscured by more aggregate-level considerations. The developmental variation in frequent binge drinking during the transition to young adulthood reflects systematic variation in success and difficulties with negotiating the transition. PMID- 8709589 TI - A longitudinal study of parenting as a protective factor for children of alcoholics. AB - OBJECTIVE: This study examined whether maternal parenting behaviors might serve to protect, or buffer, a child from the potentially negative effects associated with an alcoholic father. METHOD: This hypothesis was tested with a community sample of adolescent children of alcoholics and a demographically matched comparison group of children with nonalcoholic parents (total N = 278, 55% male). Three dimensions of parenting were considered: monitoring of child behavior, consistency of discipline, and social support. These dimensions were used in both cross-sectional and longitudinal regression analyses to predict child externalizing symptomatology, alcohol use and drug use. RESULTS: Cross-sectional results supported independent effects of parenting on child outcomes, but produced limited support for the buffering hypothesis. Longitudinal analyses revealed no prospective effects of parenting and no support for the buffering hypothesis. CONCLUSIONS: Findings suggest that both parents influence child development outcomes, but that the influence of one parent does not depend upon the influence of the other parent. PMID- 8709590 TI - Conduct disorder among adolescent alcohol and drug abusers. AB - OBJECTIVE: Delinquent behavior is prevalent among adolescents who abuse alcohol and other drugs. The present study examined the extent to which conduct disorder type behavior predates substance use involvement among 166 adolescents in treatment for alcohol and other drug abuse, and the prognostic significance of conduct disorder behaviors for the clinical course of teens during the first 2 years following treatment. METHOD: This was prospective longitudinal study of 166 alcohol and other drug abusing adolescents, 67 girls and 99 boys, averaging 15.9 years of age. Consecutive admissions were recruited from two adolescent inpatient alcohol and drug treatment facilities. Subjects were interviewed during treatment and again at 6 months and 1 and 2 years posttreatment. RESULTS: Whereas the vast majority (95%) of teens entering alcohol and drug abuse treatment programs displayed a history of conduct disorder type behavior, only 47% met DSM-III-R criteria when behaviors directly or indirectly related to alcohol and drug involvement were excluded. Boys displayed a greater number of conduct disorder behaviors than girls, but the relative incidence of these behaviors was comparable across sexes. A history of conduct disorder independent of alcohol and drug involvement was related to both greater posttreatment alcohol involvement and a later diagnosis of antisocial personality disorder. CONCLUSIONS: This study shows that a significant portion of the delinquent behaviors found in adolescents treated for alcohol and drug abuse is secondary to involvement with alcohol and other drugs. A primary diagnosis of conduct disorder was found to be associated with poorer clinical course in the first 2 years following treatment. PMID- 8709591 TI - Sources of alcohol for underage drinkers. AB - OBJECTIVE: We assessed the extent to which various sources of alcoholic beverages are used by early, middle and older teenagers, the extent to which teenagers perceive alcohol to be available from different sources and factors related to use of alternative sources. METHOD: Ninth graders (n = 2,269), twelfth graders (n = 2,377) and youth aged 18-20 (n = 1,738) were surveyed in 15 upper-midwestern communities using a nested cross-sectional design. Analyses were based on mixed model regressions, with both the individual and community treated as random effects, taking into account the intraclass correlation for each dependent variable. Analyses focused on current (last 30 day) drinkers to avoid recall bias for drinking events in the distant past. RESULTS: A person aged 21 or over was the most common source of alcohol for current drinkers in all three age groups; 46% of 9th graders, 60% of 12th graders and 68% of those aged 18-20 obtained alcohol from a person age 21 or older for their last drinking occasion. A person under age 21 was the second most prevalent source for 9th and 12th graders (29% of each group obtained alcohol from someone under age 21 for their last drinking occasion). A commercial outlet was the second most prevalent source for those aged 18-20 (14% obtained alcohol directly from an outlet for their last drinking occasion). CONCLUSIONS: Perceived availability, binge drinking and several demographic characteristics were independently related to source of alcohol used, and age interacted with several factors in predicting source of alcohol. We conclude that increased attention to sources of alcohol used by youths would facilitate efforts to reduce underage drinking. PMID- 8709592 TI - Nurses as foster parents of medically fragile children. PMID- 8709593 TI - UN Conference on Population and Development, Cairo, Egypt. PMID- 8709594 TI - Transforming nursing's vision: history, holism, and the health care system. PMID- 8709595 TI - A partnership for health between nursing faculty/students and community. PMID- 8709596 TI - Nursing program at Midway College develops partnership with Vietnam. PMID- 8709597 TI - What is a certified registered nurse anesthetist? PMID- 8709598 TI - Making the difference. PMID- 8709599 TI - Striving for a research based practice in a service setting. PMID- 8709600 TI - . . . let it begin with me. PMID- 8709601 TI - 51 cases of gastric cancer treated by the turbidity descending and stasis resolving method. PMID- 8709602 TI - Observation of curative effect of lipid lowering and fat restraining tablets on complicated obesity. AB - Lipid lowering and fat restraining tablets (LLFR tablets), or tablets of alcoholic extract of rheum, were found to have good curative effect in 64 cases of obesity complicated with hypertension, lipemia and paramenia. The course of treatment is three months. Slight abdominal pain occurred in a few cases after administration, but with spontaneous subsidence. PMID- 8709603 TI - 280 cases of hyperplasia of mammary gland treated by ru he chong ji. AB - 280 patients with hyperplasia of mammary gland were treated by Ru He Chong Ji, a breast nodule granular dissolvable agent. Thirty-one were cured, 117 markedly improved, 119 improved, and 13 showed no change. The rate of cure and marked effectiveness was 52.86%, with a total effective rate of 95.36%. The agent is also effective for treatment of depression of liver-energy and menoxenia. PMID- 8709604 TI - Guyin decoction in the treatment of immuno-sterility and its effect on humoral immunity. AB - 60 cases of immuno-sterility (treated group) were treated with Guyin Decoction, and were compared with 25 control cases treated by Western medicine. The results showed that in the treated group 19 cases were cured, 32 improved and 9 failed respond, while in the control group 6 were cured, 8 improved and 11 failed. The decoction had significant effect on serum IgA and IgG. PMID- 8709605 TI - The treatment of pain in bone metastasis of cancer with the analgesic decoction of cancer and the acupoint therapeutic apparatus. AB - The pain in 286 cases of bone metastasis of cancer was treated with an analgesic decoction of herbal drugs in combination with the Acupoint Therapeutic Apparatus that generated low frequency electric impulses into the acupoints. This series of patients comprised 104 cases of medium pain and 182 cases of severe pain, which was practically or completely relieved in 17 or 29 minutes respectively after the treatment; the total effective rate was 74.2% for 212 cases, and the duration of analgesic effect lasted 2.7 hours to 5.2 hours, averaging 3.6 hours. The cases that needed considerable dosages of analgesic tablets or sedatives markedly reduced their requests of such drugs. Determination of bone marrow stem cells indicated that the herbal treatment produced effects of promoting and protecting the stem cells. The rate of lymphocyte transformation was 45-76% before the treatment and rose to 57-96% after the treatment, demonstrating an increase of 15.2%, which was statistically significant (P < 0.001). The analgesic effect of the decoction was not correlated to the age and sex of the patients or the variety of cancer, except that it was not effective for uterine carcinoma. The main side effects included nausea, vomiting, somnolence, and numbness of the tongue. PMID- 8709606 TI - Experience of Prof. Wei Jia in using acupoint fengchi. PMID- 8709607 TI - Differential TCM treatment of anovulatory dysfunctional uterine bleeding. PMID- 8709608 TI - 38 cases of hysteromyoma treated with tumor-resolving decoction. PMID- 8709609 TI - Controlling infantile AIDS with natural remedies--a Romanian case study. PMID- 8709610 TI - Periomarthritis treated with pain point pressure in combination with local exercises. PMID- 8709611 TI - External approach to the treatment of pediatric asthma. PMID- 8709612 TI - Studies of qingyangshen (I): Differential expression of hippocampal c-fos proto oncogene during kainic acid induced acute and chronic seizures. AB - We previously reported that Qingyangshen (QYS), a traditional Chinese medicine with antiepileptic property, had therapeutic effect on kainic acid (KA) induced experimental seizures (see JTCM 13 (4): 281-286, 1993 for reference). To investigate the mechanisms underlying the anticonvulsant action of QYS, we analyzed the modulatory effect of QYS on rat hippocampal c-fos proto-oncogene expression during KA-induced epileptogenesis in this and the following paper. The expression of hippocampal c-fos gene during KA-induced seizures were examined first. Rats were intraperitoneally injected with kainic acid (KA, 12 mg/kg) and hippocampal c-fos mRNA level was determined by Northern blot analysis during both acute (within one day after KA injection) and chronic (15 days after KA treatment) seizures. A mild increase in hippocampal c-fos mRNA level was observed 30 min after KA injection (being 1.98 +/- 0.70 times of control level), which corresponded temporally to the occurrence of the first limbic seizures. There was an 11.02 +/- 3.33 fold maximal induction of c-fos mRNA at 2 h after KA administration, which remained relatively stable even when behavioral seizures continued to aggravate 4 h after KA treatment. Twelve hours after KA administration, c-fos mRNA in the hippocampus returned to control level when acute seizures began to gradually disappear. In contrast, the induction of hippocampal c-fos mRNA during chronic seizures was inhibited significantly, being reduced by 55.40% compared to control. This suggested that c-fos is in different functional states at acute and chronic stages of epileptogenesis induced by KA. PMID- 8709613 TI - Clinical observations on the treatment of gastrointestinal disorders by massotherapy. PMID- 8709614 TI - Advances in studies on acupuncture abstinence. AB - Acupuncture abstinence includes mainly abstaining from smoking, drinking and drug using by means of acupuncture therapy. In this paper, the curative effect, the stimulation methods, the therapeutic course, the psychological, demography and other factors on the therapeutic effect, and the progress in acupuncture abstinence are briefly introduced, and its development tendency is prospected. It is found that the therapeutic effects of acupuncture abstinence are not lower than those of other abstinence therapies. It is simple and easy to be performed, with little pain, little injury and little side-effects, inhibiting withdrawal syndrome, thus well accepted by the patients. It could be used if other withdrawal therapies show no apparent therapeutic effects. PMID- 8709615 TI - Human acute myeloid leukemias may be etiologically associated with new human retroviral infection. AB - The etiology of human acute myeloid leukemias (AML) remains uncertain. In order to examine the possibility of retroviral etiology in AML, we determined retroviral antigens related to HTLV-I in leukemic cell samples from 32 AML cases and peripheral blood mononuclear cells (PBMNCs) from 20 healthy donors by D-IGSS with high sensitivity and specificity, reverse transcriptase (RT) activity by a simple and sensitive non-radioisotopic RT assay, and retroviral particles by electron microscopy. The HTLV-I-related antigens were detected in 50.0% (16/32) of fresh leukemic cell samples and 87.5% (28/32) of cultured leukemic cell samples. The HTLV-I-related antigen-positive cells in fresh and cultured leukemic samples were 10.2% and 52.8%, respectively. Both frequency and level of HTLV-I related antigens in cultured samples were much higher than in fresh samples. In contrast, no HTLV-I-related antigens were found in normal hematopoietic cells from 20 healthy donors. Further study results show that RT activity was detected not only in HTLV-I-related antigen-positive samples, but was also well correlated with the level of HTLV-I-related antigens in these samples, and preferred Mn+2 to Mg+2 as a cation. Moreover, typical retroviral particles were localized in most cultured HTLV-I-related antigen-positive samples by immunoelectron microscope. These data suggest that human acute myeloid leukemias may be etiologically associated with new human retroviral infection. PMID- 8709616 TI - HTLV infection and hematologic malignancies. PMID- 8709617 TI - Analysis of the erythropoietin receptor gene in patients with myeloproliferative and myelodysplastic syndromes. AB - The human erythropoietin receptor (EpoR) gene has been cloned and characterized. Very few EpoR genetic abnormalities have been reported so far. Polycythemia vera (PV) is characterized by low/normal serum erythropoietin (Epo) levels with proposed Epo hypersensitivity. Myelodysplastic syndromes (MDS) are characterized by refractory anemia with variable serum Epo levels. Several reports have suggested EpoR abnormalities in both types of stem cell disorders. We analyzed DNA obtained from peripheral blood mononuclear cells of seven healthy controls, 20 patients with myeloproliferative disorders (MPD, 11 patients with PV, five agnogenic myeloid metaplasia with myelofibrosis, four essential thrombocytosis) and eight patients with refractory anemia with ringed sideroblasts (RARS), an MDS variant. The DNA was digested with four restriction enzymes (BamHI, Bgl II, Sacl and HindIII), followed by Southern blot, using a 32P radiolabeled probe, containing 1.5 kb of the human EpoR cDNA. All 20 MPD patients and seven out of the eight MDS patients demonstrated a restriction pattern which was identical to the seven normal controls, as well as to the erythroid cell line K562, and also consistent with the expected restriction map, for all four enzymes tested. One RARS patient had a normal pattern with three enzymes but a different one with HindIII. The HindIII 12 kb large band was replaced by a faint 12 kb band and a new (about 9 kb) band appeared. The EpoR restriction map and the normal pattern obtained with the other three enzymes suggest that this patient has a 3 kb upstream deletion in one allelic EpoR gene. The same molecular pattern was detected in the patient's sister, who suffers from anemia with mild bone marrow (BM) dyserythropoiesis and plasmacytosis. Northern blot analysis showed that the patient's BM RNA carried normal EpoR message. This familial pattern may represent polymorphism. However, the patient's very high serum Epo level, her resistance to treatment with recombinant Epo, and the abnormally low growth rate of in vitro erythroid cultures, suggesting poor response to Epo in this MDS patient as well as the hematological abnormalities in her sister, support the speculation that the different EpoR gene might serve as a genetic predisposing marker and potentially could be involved (probably via post-transcriptional mechanisms and by an interaction with other factors or cytokines) in the pathogenesis. Our data suggest that the EpoR is intact in MPD and in most patients with RARS. One RARS patient had a familial different genetic structure, which could represent polymorphism. However, we can speculate also that it might be involved in the pathogenesis of the disease. PMID- 8709618 TI - Usefulness and limitations of serum and urine lysozyme levels in the classification of acute myeloid leukemia: an analysis of 208 cases. AB - The revised French-American-British (FAB) classification system for acute myeloid leukemia (AML) recommends the determination of serum lysozyme (SL) or urine lysozyme (UL) levels as an aid in distinguishing acute myeloblastic leukemia with maturation (FAB M2) from acute myelomonocytic leukemia (M4). We reviewed retrospectively 208 cases of adult leukemia in which SL and/or UL were obtained. Elevated lysozyme levels were not found in any of the M0, M3, or M7 cases, but were increased (false positive) in three (14%) M1 cases, 18 (19%) M2 cases and one (20%) M6 case. Although a UL value in excess of 3x normal was found in most cases of AML M4 and M5, only five (11%) M4 cases and three (20%) M5 cases had SL elevations of this magnitude. Lysozyme levels need to be interpreted in conjunction with other parameters for FAB classification. PMID- 8709619 TI - The effect of bcr-abl antisense oligonucleotide on DNA synthesis and apoptosis in K562 chronic myeloid leukemia cells. AB - Mutations in oncogenes have traditionally been viewed as inducing malignancy by causing excessive cell division. However, an additional possible tumorigenic mechanism is inhibition of normally occurring apoptosis. We have studied the mechanism of action of bcr-abl in chronic myeloid leukemia (CML) by inhibiting its expression using antisense oligonucleotides. K562 cells, derived initially from a patient with CML, were incubated with 16 microM 3',5'-capped bcr-abl antisense phosphodiester 18mer targeting the bcr-abl junctional sequence. Antisense reduced cell number by day 5 by 44% +/- 2.5% S.E. compared to nonsense or no-oligomer controls. Compared to nonsense oligomer, antisense oligomer reduced [3H]thymidine incorporation by only 13% +/- 1%. By the more reliable bromodeoxyuridine incorporation method, antisense had no inhibiting effect on DNA synthesis. In contrast to its minimal effect on DNA synthesis, antisense had a large effect on apoptosis. At day 4, after 3 days of oligomer treatment, antisense increased the proportion of cells with less than 2 N DNA 2.5 +/- 0.3 fold compared to nonsense, as revealed by analysis of DNA distribution following propidium iodide-staining. After 3 days of oligomer treatment and 24 h of serum deprivation, antisense increased the proportion of cells with less than 2 N DNA even more, over 3.1 +/- 1.1-fold compared to nonsense. Because CML cells are resistant to the induction of apoptosis (as judged by DNA laddering on electrophoresis, which requires double-stranded breaks), we also assayed the binding of terminal deoxynucleotidyl transferase (TdT), which requires only single-stranded DNA breaks. Antisense treatment for 3 days increased TdT binding at day 4 by 16.4 +/- 8.7-fold. We conclude that, in CML, bcr-abl may lead to the accumulation of myeloid cells to a greater extent by inhibiting apoptosis than by increasing cell division. This bcr-abl induced inhibition of apoptosis may thwart chemotherapy and foster the accumulation of further mutations leading to the development of the blastic phase of the disease. PMID- 8709620 TI - Influence of rhGM-CSF on Ara-C sensitivity of patients with acute myeloid leukemia in relapse: a flow cytometry study. AB - Twenty-three patients with acute myelogenous leukemia (AML) in first relapse were treated with high-dose cytosine-arabinoside (Ara-C) and amsacrine or idarubicin. To prime the cells, the patients were given rhGM-CSF. We studied the influence of 48-h infusion of rhGM-CSF on proliferation and Ara-C sensitivity of leukemic cells both ex vivo and in vitro. We found that a 48-h infusion of rhGM-CSF increased both white blood cell counts and peripheral blood blast cell percentages. Using a Bromodeoxyuridine/DNA (BrdUrd/DNA) staining in flow cytometry, we found an non-constant increase in cells in the S-phase. Ex vivo 48 h culture of leukemic cells with or without rhGM-CSF, with or without other hematopoietic growth factors (HGFs), showed a greater increase of the cells in the S-phase with GF but no correlation with the ex vivo results. We used a method of quantitation of the DNA synthesis previously described (Lacombe F., et al. (1992) Cytometry 13, 730) to monitor the Ara-C sensitivity of the cells in S phase before and after 48-h infusion with rhGM-CSF. We observed a great variation in the Ara-C sensitivity of the leukemic cells before and after infusion with rhGM-CSF from one patient to another. The BrdUrd/DNA method seems a convenient method to study the influence of HGFs on Ara-C sensitivity of the patients. PMID- 8709621 TI - Angelmicin B, a new inhibitor of oncogenic signal transduction, inhibits growth and induces myelomonocytic differentiation of human myeloid leukemia HL-60 cells. AB - Angelmicin B is a new microbial substance which inhibits src tyrosine kinase activity and oncogenic signal transduction. We investigated the effect of angelmicin B on the proliferation and differentiation of the HL-60 human myeloid leukemia cell line. Angelmicin B caused the dose-dependent inhibition of cell proliferation and induction of differentiation along the myelomonocytic pathway, as determined by morphological changes, nitroblue tetrazolium (NBT) reduction, and non-specific esterase and lysozyme activities at concentrations ranging from 0.1 to 0.5 microgram/ml. Also, it induced significantly the differentiation of mouse myeloid leukemia M1 cells. A similar concentration of angelmicin B inhibited the growth of the myeloid leukemia cell lines K562, HEL, KU812, ML-1, U937 and THP-1, but did not induce differentiation of these cells significantly. The differentiation of HL-60 cells was enhanced by combined treatment with angelmicin B and 1 alpha, 25-dihydroxyvitamin D3 (VD3), retinoic acid or tumor necrosis factor-alpha (TNF alpha). Angelmicin analogs (A1, A2, B, C and D) had almost equivalent effects on the differentiation of HL-60 cells, although angelmicins C and D inhibited src tyrosine kinase activity less than the other analogs. The effective concentrations of angelmicin B in src kinase inactivation was about 100-fold higher than those required for the growth inhibition and differentiation induction. These findings indicate that the differentiation inducing activity of angelmicins is not associated with their src kinase inhibiting activity, and may be associated with the modulation of other signal pathway(s). PMID- 8709622 TI - Retinoid-regulated expression of BCL-2 and tissue transglutaminase during the differentiation and apoptosis of human myeloid leukemia (HL-60) cells. AB - Retinoids induce terminal differentiation and subsequent apoptosis in the human myeloid leukemia (HL-60) cell line. We have previously shown that in HL-60 cells, ligand activation of retinoic acid receptors (RARs) is sufficient to induce differentiation but ligand activation of retinoid X receptors (RXRs) is necessary for the retinoid-induced apoptosis of these cells. In the present studies we have characterized the effect of retinoids on the expression of two apoptosis-linked gene products, BCL-2 and tissue transglutaminase. BCL-2 is a membrane-associated protein whose expression has been linked to the suppression of apoptosis in many cells. Tissue transglutaminase is a protein cross-linking enzyme that accumulates in many cells undergoing apoptotic cell death. Our data suggest that ligand activation of RARs in HL-60 cells results in a global suppression of BCL-2 expression whereas ligand activation of both RARs and RXRs triggers the selective accumulation of tissue transglutaminase in the apoptotic HL-60 cells. PMID- 8709623 TI - CD40 ligand triggers interleukin-6 mediated B cell differentiation. AB - Interleukin-6 (IL-6) is the major cytokine to date mediating antigen (Ag)- or mitogen-driven B cell differentiation. Recently, CD40 ligand (CD40L), with the co stimulatory cytokines IL-4 and IL-10, has been shown to trigger immunoglobulin (Ig) secretion and class switching. In the present report, we have examined the role of IL-6 in mediating B cell differentiation and Ig secretion triggered with CD40L and/or these cytokines. Culture of splenic B cells with CD40L triggered (1) significant (5.4-fold) increases in IL-6 secretion; (2) differentiation, evidenced by sequential loss of B cell (CD20, CD21) and acquisition of plasma cell (CD38, PCA-1) surface antigens (Ags); and (3) Ig secretion. Interleukin-4 increased both IL-6 and IgG secretion stimulated by CD40L. Interleukin-10+ CD40L triggered 100-fold increments in IgG, IgA and IgM secretion, but IL-10 suppressed IL-6 secretion triggered with CD40L +/- IL-4. Exogenous IL-6 can further increase IgG secretion induced by CD40L + IL-10; moreover, the anti-IL-6 monoclonal antibody partially blocked IgG secretion triggered by CD40L +/- IL-4 or IL-10. Finally, IL-10 suppressed differentiation of B cells induced by CD40L. These studies suggest that CD40L augments Ig secretion in at least two mechanisms: by triggering IL-6 secretion and related differentiation, and by priming B cells for responsiveness to IL-10. PMID- 8709624 TI - Oligoblastic leukaemia with (8;21) translocation and haemophagocytic syndrome and granulocytic cannibalism. AB - We report a 47-year-old man with oligoblastic leukaemia (8;21) translocation, phenomenon of cannibalism by granulocytic cells and haemophagocytic syndrome. The patient responded to intensive chemotherapy with disappearance of haemophagocytosis, granulocytic and histiocytic. We conclude that: (1) granulocytic cannibalism and haemophagocytic syndrome can be unusual myelodysplastic features; (2) the oligoblastic leukaemia with presence of cytogenetic abnormalities related to AML in young patients are probably more close to acute leukaemia than to myelodysplastic syndrome. PMID- 8709625 TI - BCR/ABL and leukemia. AB - This review focuses on the role of the chimeric BCR/ABL gene in leukemia development. First, we discuss and update knowledge regarding the molecular biology of BCR/ABL. We then review data regarding transforming activity of BCR/ABL. Third, we discuss the complex interactions between BCR/ABL and leukemia phenotype. We conclude with a brief discussion of possible therapeutic implications of these data. PMID- 8709626 TI - What is a 'step' in a multi-step pathogenesis of leukemia? PMID- 8709627 TI - Responsiveness of AML cells to recombinant hematopoietic growth factors: relationship with morphological and immunological characteristics of blast cells. PMID- 8709628 TI - Telomeres and telomerase in human leukemias. AB - There is increasing evidence supporting the hypothesis that telomere shortening both in vitro and in vivo, is the clock that counts cell divisions and determines the onset of cellular senescence. Cells that overcome the normal senescence mechanisms do so by stabilizing telomere length, probably due to the activity of telomerase, a ribonucleoprotein enzyme that synthesizes telomeric repeats. Most human primary tumors contain telomerase, while the cells of most normal tissues lack this activity. A hypothesis gaining prominence is that the activation of telomerase is necessary for the sustained growth of most solid tumors. Since normal hematopoietic stem cells and some of their progeny already express telomerase activity, it is important to consider whether or not telomere shortening and telomerase activity play any role in cancer progression in various forms of leukemia. This review includes a discussion of the utility of telomere length and/or telomerase activity measurements in the diagnosis and prognosis of leukemia as well as the potential value of antitelomerase therapy for the leukemias. PMID- 8709629 TI - Differentiating therapy in acute myeloid leukemia. AB - Differentiating therapy is a new antineoplastic strategy which has received increasing attention due to the remarkable activity of the vitamin A derivative, all-trans retinoic acid (ATRA) in patients with acute promyelocytic leukemia (APL). Although it has been known for years that a variety of agents, including retinoids, could induce leukemic cells to differentiate in vitro, it was not until the initial report from Shanghai in 1988 that laboratory studies translated into clinical activity and benefit in patients. Since this initial report, a number of studies have confirmed that the majority of patients with both newly diagnosed and previously chemotherapy-treated patients with APL achieve complete remission (CR) with ATRA. In addition, the characteristic life-threatening coagulopathy resolves quickly. Several limitations to this approach have emerged, including the development of retinoid resistance, hyperleukocytosis and the retinoic acid syndrome, a constellation of findings including unexplained fever, fluid retention, pleuropericardial effusions and pulmonary infiltrates. Although ATRA is very effective in inducing CR, its benefits compared to conventional chemotherapy are only now being addressed. The first prospective randomized trial comparing ATRA plus chemotherapy to chemotherapy alone was terminated early because of an improved event-free survival for patients receiving ATRA. The benefit was attributable to a difference in relapse rate. A large, intergroup, prospective, randomized trial comparing conventional chemotherapy to ATRA for induction and ATRA to observation for maintenance has recently completed accrual and will provide insight into the emerging role of ATRA in patients with APL. ATRA represents the first example of a specific form of antileukemic therapy targeting a specific genetic abnormality and may serve as a paradigm for the development of differentiating therapy for patients with other hematologic malignancies. PMID- 8709630 TI - Intensive chemotherapy in childhood myelodysplastic syndrome. A comparison with results in acute myeloid leukemia. AB - Myelodysplastic syndrome (MDS) in children is often considered as a variant of acute myeloid leukemia (AML) and frequently treated as such. However, there are very few reported data on the outcome following AML treatment. We analyzed 20 consecutive cases of de novo MDS treated in Denmark according to the NOPHO AML protocols. The results were compared with those obtained in 31 children with de novo AML treated with the same protocols, and with the outcome in 10 children with MDS who received allogeneic bone marrow transplantation (BMT) without prior AML therapy. Distinction between MDS and AML was made morphologically according to the FAB criteria. All children were followed for at least 37 months. The proportion of complete remission in MDS and AML was 35 percent vs 74 percent. (P = 0.005), resistant disease 25 percent vs 10 percent (P = 0.14), death in cytopenia 40 percent vs 16 percent (P= 0.06), and 3-year survival 15 percent vs 35 percent. (P = 0.11), respectively. Duration of treatment-related cytopenia was similar in MDS and AML, except for a longer period of leukopenia in MDS following the second course of induction. Seven of 10 MDS children receiving BMT without prior chemotherapy are long-term survivors. Our data suggest that conventional AML regimens are associated with a low rate of complete remission, a high risk of death in cytopenia, and a limited curative potential in childhood MDS. Allogeneic BMT was in contrast associated with a high survival rate. BMT may, at least in some patients, be performed successfully without prior induction chemotherapy. The different response to therapy in MDS and AML may reflect fundamental biological differences between the two conditions. PMID- 8709631 TI - Multidrug resistance-1 (MDR1) expression and functional dye/drug efflux is highly correlated with the t(8;21) chromosomal translocation in pediatric acute myeloid leukemia. AB - Resistance to chemotherapy is a major problem in acute myeloid leukemia (AML). An important resistance mechanism in adult AML is active drug efflux mediated by the multidrug resistance protein-1 (MDR1). To determine if MDR1 is important in childhood AML, we examined MDR1 expression and functional dye/drug efflux in 20 pediatric/adolescent AML patients; results were correlated with cytogenetics and clinical outcome. Using flow cytometry, MDR1 protein expression on the leukemic blasts was measured with the antibody MRK16, while efflux was measured by extrusion of the fluorescent dye DiO(C2)3 in the presence/absence of cyclosporin A (CsA). Six of 20 cases expressed MDR1. While all six MDR1+ cases were efflux+, three of 14 MDR1- cases also demonstrated efflux. Both MDR1 and efflux were strongly correlated with the t(8;21). All six MDR1 +/efflux+ cases and 2/3 MDR1 /efflux+ cases had a t(8;21), while no MDR1-/efflux- cases had a t(8;21) (P < 0.0005). This correlation between MDR1, efflux, and the t(8;21) in pediatric AML was not found in 11 adult t(8;21) cases similarly studied. Although the clinical relevance of MDR1 in pediatric AML awaits larger studies, our results suggest a biologic subset of pediatric AML patients may benefit from regimens which include MDR1-reversing agents or non-MDR1 substrates. PMID- 8709632 TI - Cytogenetic findings in acute biphenotypic leukaemia. AB - We have studied the immunological and cytogenetic features of 26 patients with acute leukaemia classified as biphenotypic according to a scoring system based on the number and lineage specificity of antigens expressed on the blast cells. The series included 19 adults (age >15 years) and seven children. The cases were distributed in four immunophenotypic groups: (1)coexpression of myeloid and B antigens, 18 cases (69 percent);(2)myeloid and T cell antigens, six (23 percent); (3) one case with trilineage differentiation; and (4) one case with coexpression of both B and T cell antigens. Cytogenetic analysis revealed a normal karyotype in four cases (15 percent) and abnormal clones in 22 (85 percent). Eight patients had the Philadelphia (Ph) translocation, t(9;22)(q34;q11), (31 percent), three cases had structural aberrations of 6q and two had 11q23 rearrangements, one with t(11;19) and a second with t(4;11); the other eight cases had different alterations including t(9;12)(q1;q1), t(8;21)(q22;q22), t(2;7) (p1?3;q3?4), t(7;12)(q11;p11), hyperdiploidy and other structural abnormalities. The chromosomal rearrangements in children were characterised by abnormalities of 11q23 in two cases and the Ph translocation in three. Our data indicate that biphenotypic features are common in cases presenting with t(9;22) as the eight cases included here represent 47 percent of all cases of Ph+ve acute leukaemia studied in our Institution. Biphenotypic acute leukaemias comprise a heterogeneous group of leukaemias involving pluripotent stem cells. Cytogenetic studies are essential in characterising these cases as they will disclose several poor prognosis chromosome aberrations of which the Ph chromosome is the most frequent. PMID- 8709633 TI - Fifty-one patients with acute myeloid leukemia and translocation t(8;21)(q22;q22): an additional deletion in 9q is an adverse prognostic factor. AB - The translocation t(8;21)(q22;q22) occurs in 6 to 12 percent of patients with AML, and usually predicts a good response to chemotherapy with a high remission rate and a relatively long median survival. The influence of additional chromosome aberrations on the clinical outcome of patients with t(8;21) is unclear. We analyzed 51 cases of acute myeloid leukemia carrying a translocation t(8;21)(q22;q22); 23 female and 28 male patients. The complete remission rate was 92 percent and median overall survival was 52.4 months. The median overall survival of female patients was significantly worse than of male patients (37.2 months vs not reached, P = 0.025). Additional chromosome aberrations were detected in 41 patients at diagnosis (80 percent), 31 (61 percent) had lost a sex chromosome, seven (14 percent) showed a partial deletion of the long arm of chromosome 9 and in three patients (6 percent) a gain of chromosome 8 was observed. Whereas the loss of a sex chromosome had no influence on prognosis, a partial deletion of the long arm of chromosome 9 was an unfavorable prognostic factor. The median overall survival of the seven patients with del(9q) was only 12.5 months and thus significantly shorter than in patients with only t(8;21) or with t(8;21) and additional sex chromosome loss (median survival not reached: P = 0.0010). PMID- 8709634 TI - Molecular analysis of the PML/RAR alpha chimeric gene in pediatric acute promyelocytic leukemia. AB - Acute promyelocytic leukemia (APL) is characterized cytogenetically by the t(15;17)(q22;q11-21) translocation. To compare molecular events among pediatric and adult APL cases, we designed two sets of oligonucleotide primers using published cDNA sequence for PML/RAR alpha fusion transcripts, and undertook reverse transcriptase-polymerase chain reaction (RT-PCR) analysis of 22 US pediatric cases of APL. PML/RAR alpha fusion transcripts were detected in all APL cases, including two cases lacking cytogenetic evidence of t(15;17). Breakpoint usage in PML was determined using a combination of PCR amplification with differing 5' primers, junction-specific probes, and sequence analysis in selected cases. Consistent with previously published data, case analysis demonstrated fusion products resulting from three breakpoint cluster regions (bcr) in PML, and a single breakpoint region in intron 2 of RAR alpha. Transcripts resulting from breakpoints in bcr1 were detected in 59 percent of cases, bcr2 in 27 percent and bcr3 in 14 percent. This distribution is dissimilar to that observed in adults, where bcr2 comprises a lesser and bcr3 a greater portion of cases. These results suggest that the pathogenesis of the t(15;17) in APL may differ among patient sets. RT-PCR with these primer sets is a reliable method for detecting PML/RAR alpha chimeric transcript in t(15; 17)-containing APL. PMID- 8709635 TI - Frequency and clinical significance of the MLL gene rearrangements in infant acute leukemia. AB - We have analyzed the frequency and clinical significance of the MLL gene rearrangements in 42 cases of infant acute leukemias; including 37 cases of acute lymphoblastic leukemia (ALL) and five cases of acute myeloid leukemia (AML). MLL gene rearrangements were found in 27 of the 37 ALL cases (73 percent), and in all five AML cases. Cytogenetic studies showed 11q23 abnormalities in 24 of 27 ALL cases with MLL gene rearrangements. MLL gene rearrangements were significantly correlated with absence of CD10 expression and poor prognosis, but not with age under 6 months, hyperleukocytosis, myeloid-associated antigen expression, or CNS leukemia. The 3-year overall survival rate for ALL cases with MLL gene rearrangements was 5.3 +/- 5.2 percent, compared with 88.9 +/- 10.5 percent for cases with germline MLL (P=0.0001). Absence of CD10 expression was also associated with poor prognosis (9.9 +/- 6.6 percent vs 85.7 +/- 13.2 percent, P = 0.0003). Of the five AML cases, three have remained alive for 27 months to 67 months. These findings suggest that infant ALL with MLL gene rearrangement is strongly associated with poor prognosis. We consider that infant ALL should be treated on different chemotherapy protocols according to the presence or absence of MLL gene rearrangement. PMID- 8709636 TI - Effects of IL-10 and IL-4 on LPS-induced transcription factors (AP-1, NF-IL6 and NF-kappa B) which are involved in IL-6 regulation. AB - Interleukin-10 (IL-10), like IL-4, is known to inhibit cytokine expression in activated human monocytes. We showed that both IL-10 and IL-4 inhibit LPS-induced IL-6 mRNA and protein expression by inhibiting the transcription rate of the IL-6 gene. The strong inhibition of the IL-6 transcription rate prompted us to study the effect of IL-10 and IL-4 on the expression of transcription factors. We questioned whether or not IL-10 and IL-4 affected the expression of transcription factors that are known to be involved in the control of the IL-6 transcription rate, namely activator protein-1 (AP-1), nuclear factor IL-6 (NF-IL6), and nuclear factor kappa B (NF-kappaB). In electrophoretic mobility shift assays (EMSAs) we showed that IL-10 and IL-4 inhibited LPS-induced AP-1 binding activity. The inhibiting effect of IL-4 was slightly more pronounced than that of IL-10. Downregulation of LPS-induced AP-1 was accompanied, and thus possibly explained, by a reduced expression at mRNA level of the two major components of the AP-1 complex, namely c-fos and c-jun as determined by Northern experiments. Binding activity of NF-IL6 was also strongly inhibited by IL-4 whereas IL-10 showed no effect. NF-IL6 mRNA levels were not affected by IL-10 or IL-4, suggesting that IL-4 affects binding activity of preexisting NF-IL6. Neither IL 10 nor IL-4 inhibited LPS-induced NF-kappa B binding activity. In agreement with this finding, Northern experiments where p65 and p105 mRNA levels were determined, demonstrated that expression of these components of the NF-kappa B transcription factor were not affected by IL-10 or IL-4. Furthermore, neither IL 10 nor IL-4 showed any effect on I-kappa B mRNA expression as determined by Northern experiments. Thus, IL-10 and IL-4 similarly affect IL-6 expression. However, for IL-4 this was accompanied with a reduction of AP-1 and NF-IL6 binding activity whereas IL-10 only inhibited AP-1 binding activity. PMID- 8709637 TI - Interleukin-7 signaling in human B cell precursor acute lymphoblastic leukemia cells and murine BAF3 cells involves activation of STAT1 and STAT5 mediated via the interleukin-7 receptor alpha chain. AB - Interleukin-7 (IL-7) stimulates the proliferation of normal and leukemic B and T cell precursors and T lymphocytes. Activation of the JAK/STAT pathway has been implicated in IL-7R signaling. We investigated which STAT complexes are formed upon stimulation of B cell precursor acute lymphoblastic leukemia (BCP-ALL) cells with IL-7. Gel retardation assays with STAT-binding oligonucleotides showed that IL-7 induces the formation of two major STAT complexes in BCP-ALL cells. Supershifts with anti-STAT antibodies identified these as STAT1 and STAT5 complexes. This pattern of STAT activation was seen in all BCP-ALL cases that respond to IL-7 in proliferation assays. IL-7 also induced STAT/DNA binding in BCP-ALL cases that failed to proliferate in response to IL-7, suggesting that the ability of IL-7R to activate the JAK/STAT pathway per se is not sufficient for proliferation induction. To determine the contribution of the cytoplasmic domain of the IL-7 receptor alpha chain (IL-7R alpha) to activation of STAT proteins, transfectants of the murine pro-B cell line BAF3 were made that express chimeric receptors consisting of the extracellular domain of human granulocyte colony stimulating factor receptor (G-CSF-R) and the transmembrane and intracellular domains of human IL-7R alpha. Activation of the chimeric G-CSF-R/IL-7R alpha with G-CSF resulted in a full proliferative response and induced the phosphorylation of JAK1 but not JAK2. Major STAT complexes activated by G-CSF-R/IL-7R alpha contained STAT1 or STAT5, while some formation of STAT3-containing complexes was also seen. These findings establish that STAT1 and STAT5, and possibly STAT3, are activated upon stimulation of precursor B cells with IL-7. The data further indicate that the IL-7R alpha chains are directly involved in the activation of JAKs and STATs and have a major role in proliferative signaling in precursor B cells. PMID- 8709638 TI - Blood cell flow cytometry in paroxysmal nocturnal hemoglobinuria: a tool for measuring the extent of the PNH clone. AB - The membrane expression of nine glycosyl phosphatidyl inositol (GPI)-linked molecules was analyzed by flow cytometry on circulating cells from 18 patients affected by paroxysmal nocturnal hemoglobinuria (PNH). The results allowed us to select CD66b, CD14, CD59, CD24 and CD59 monoclonal antibodies as the most suitable reagents for discriminating between normal and PNH cells in PMN, monocytes, RBC and B or T lymphocytes, respectively. In order to assess whether the analysis of distinct cell populations could provide differential information on the extent of the disease, we compared the proportion of residual normal cells in RBC, monocyte and PMN populations. The mean percentage of unaffected cells was higher in RBC as compared to PMN (50.5 +/- 18.7 vs 17.7 +/- 19.7, P < 0.0001). The proportion of normal PMN was, in turn, significantly greater than that of normal monocytes (17.7 +/- 19.7 vs 8.7 +/- 11.0; P < 0.05). The percentage of CD14+ monocytes was directly related to Hb concentration and platelet (Plt) count, and inversely to percent lysis at the Ham's test. The percentage of CD66b+ PMN was directly related to Plt count and Hb level, while the percentage of CD59+ RBC was associated, in an inverse fashion, only to the Ham's test. No significant correlation was found between cell marker expression and PMN count, reticulocytosis, bilirubin and serum LDH. By dividing the patients into two groups, according to high (> 10 percent) or low (< 10 percent) percentage of CD14+ monocytes, a statistical analysis showed that the main hematological parameters were significantly different. PMID- 8709639 TI - Variable expression of CD49d antigen in B cell chronic lymphocytic leukemia is related to disease stages. AB - We have investigated the expression of the dual specific adhesion molecule, VLA-4 (CD49d/CD29) on lymphocytes obtained from 62 patients with B-CLL and compared it with normal controls, patients with other hematological malignancies, and umbilical cord blood. The mean CD49d expression in patients with CLL was lower than the other group of leukemia and the CD19+, CD5+ cells of normal peripheral blood and umbilical cord blood (P < 0.001). The patients in RAI stage 0, I and II (early stage) had even lower CD49d expression, whereas patients in RAI stage III and IV (advanced stage) had relatively higher CD49d levels. In vitro adhesion of lymphocytes to fibronectin, being the extracellular matrix ligand of CD49d, was also investigated. Lymphocytes obtained from B-CLL were found to have lower adhesion to fibronectin than that from controls (P < 0.03). Furthermore, CD49d(low) B-CLL cells had lower adhesion to fibronectin, whereas CD49d(high) B CLL cells showed normal adhesion ratios (P < 0.002). Further phenotypic analyses revealed the presence of myeloid markers (CD13 and CD33) in most of the advanced stage patients, although these were negative in early stage cases. Expressions of CD11a and sIgM were also low but CD11b was relatively higher in the early stages of the disease. On the basis of these results, we concluded that early stages of CLL are correlated with the expression of CD49d(low), CD11a(low), CD11b(high), CD13-, CD33-, sIgM(low) and also had lower fibronectin adhesion, whereas advanced stages of CLL are associated with CD49d(high), CD11a(high), CD11b(low), CD13+, CD33+, SIgM(high) and show normal fibronectin adhesion. PMID- 8709640 TI - Functional role of PECAM-1/CD31 molecule expressed on human cord blood progenitors. AB - CD31/PECAM-1 (platelet endothelial cell adhesion molecule-1) is a 130 kDa integral membrane protein of the immunoglobulin gene superfamily with the distinctive feature of being expressed on several cell types associated with the vascular compartment. In the present study we report a novel, unique CD31 mAb termed IP28A which reacts with all CD34 molecule expressing hematopoietic progenitor cells and a subset of T, B and NK lymphocytes from human cord blood. Interestingly, we show that the number of CFU-GM and BFU-E was significantly augmented in cord blood progenitor cultures when purified IP28A mAb was added to rhSCF plus rhGM-CSF and rhEpo, respectively. Thus, these results are of relevance in the field of hematopoietic stem cell transplantation as they reveal an agonistic property of the IP28A/CD31 mAb on the differentiation of cord blood progenitor cells. PMID- 8709641 TI - Phenotype and progeny of primitive adherent human hematopoietic progenitors. AB - Hematopoietic progenitor cells can be classified as plastic- and stroma-adherent (P+S+), stroma-adherent (P-S+) and non-adherent (P-S-). Both P+S+ and P-S+ populations are detected in delta (delta) culture systems where they produce non adherent (P-S-) granulocyte-macrophage colony-forming cells (CFU-GM) and erythroid burst-forming units (BFU-E). Here we demonstrate that the plastic adherent progenitor cells (P delta cells) comprise 5-10 percent of the CD34+, population in adult human marrow. Moreover, they do not express CD3 or CD22 and 88 percent of them are CD38-, 88 percent are CD33- and 74 percent are HLA-DR-. Production of CFU-GM by purified plastic-adherent CD34+, adherent cells was 60 percent of the number produced by recombined CD34+, and CD34- fractions. We have shown also that the plastic-adherent P+S+ cells are the precursors of the stroma adherent P-S+ cells (S delta cells), day 21 cobblestone-area forming cells (CAFC) and cells capable of sustained hematopoiesis in a modified long-term bone marrow culture system. These observations support the primitive nature of P delta cells and establish a phenotypic sequence of plastic and stroma adherence through stroma adherence to non-adherence in hematopoietic cell development. To further investigate the relationship between P delta cells, S delta cells and long-term culture-initiating cells (LTCIC), we cultured whole mononuclear cell tractions and plastic-adherent cell-depleted mononuclear cell fractions in long-term culture and in the S delta assay. The results indicated the P delta cells were inhibited in the presence of stromal cells. PMID- 8709642 TI - Supression of telomerase activity in HL60 cells after treatment with differentiating agents. AB - The human promyelocytic leukemic HL60 cells are immortal and as such express high levels of telomerase activity. All-trans retinoic acid (ATRA) and 1 alpha, 25 dihydroxyvitamin D3 (VD3) induce differentiation of HL60 cells into CD11b+ mature granulocytes and monocytes, respectively. We studied telomerase activity after differentiation of HL60 cells. A marked inhibition of the enzyme activity was observed in the differentiated CD11b+ cells after 72-120 h treatment with either differentiating agent. In contrast, the VD3-treated CD11b- HL60 cells, which failed to undergo differentiation and human erythroleukemic cell line K562, exposed to ATRA retained high levels of telomerase activity. This finding suggests, that telomerase activity is repressed as a differentiation-associated event in HL60 cells. Our results provide the first evidence that immortal leukemic cells, like normal human cells, have a telomerase repressing mechanism which can be activated by differentiation and thus lead to the suppression of telomerase activity. This in vitro model may be useful for studies of the mechanisms controlling telomerase activity and in the search for physiological telomerase modulators. PMID- 8709643 TI - Detection of HHV-8/KSHV DNA sequences in AIDS-associated extranodal lymphoid malignancies. AB - Herpesvirus-like DNA sequences have been identified in a high proportion of both AIDS-associated and classical Kaposi's sarcoma, and in a small percentage of AIDS associated malignant lymphomas. To determine the extent of involvement of this new agent designated HHV-8 (human herpesvirus type 8) or KSHV (Kaposi's sarcoma associated herpesvirus) in human malignant lymphomas, we analyzed 24 AIDS associated lymphoid malignancies and 100 non-AIDS-associated lymphomas by PCR and Southern blot analysis. Three of 24 lymphoid malignancies from patients with AIDS demonstrated HHV-8 sequences by Southern blot and PCR analyses. The fourth was positive by PCR only. None of the non-AIDS-associated lymphomas contained HHV-8 sequences. All three Southern blot positive samples were derived from extranodal regions, two from pleural effusions, and one from a soft tissue mass in the thigh. This latter patient initially presented with a pleural effusion. The fourth PCR positive but Southern blot negative tumor was from a gingival lymphoma in a patient with a history of Kaposi's sarcoma. All tumors positive for HHV-8 were also positive for EBV. These results confirm a recent report that this novel herpesvirus may play a role in AIDS-associated lymphomas especially in those with body cavity presentation. PMID- 8709645 TI - Morphologic classification of acute myeloid leukemia: concordance among Eastern Cooperative Oncology Group investigators: a comment. PMID- 8709644 TI - Classification of acute leukemias: reply from EGIL to Dr van Dongen. PMID- 8709646 TI - HTLV-I-associated adult T cell leukemia/lymphoma in two patients from Bucharest, Romania. AB - Two middle-aged patients with T cell lymphoma, both natives of Bucharest, Romania, tested positive for HTLV-I antibodies. Malignant cells had the typical phenotype and morphology of adult T cell leukemia/lymphoma (ATL). Both cases presented with extranodal manifestation, hypercalcemia, early recurrence after initial responses to therapy, and subsequent resistance to conventional and intensified chemotherapy. Infection with HTLV-I was confirmed by PCR analyses of serial biopsies. Neither patient reported known risk factors for HTLV-I infection. This report points to the possibility that Romania may represent an endemic area for HTLV-I and should heighten the awareness towards HTLV-I infections in Romanian patients. PMID- 8709647 TI - Engraftment of human chronic lymphocytic leukemia cells in SCID mice: in vivo and in vitro studies. AB - Chronic lymphocytic leukemia (CLL) is a heterogeneous disease of the elderly which can present in one of three stages; benign, intermediate or advanced. The molecular events governing the progression of CLL are poorly understood. In order to develop model systems for predicting the aggressiveness of leukemic clones in CLL, in vivo transplantation of SCID mice with CLL cells, and the in vitro growth of CLL cells on mouse and human stromal layers, were investigated. Bone marrow or peripheral blood cells from 40 patients at different stages of CLL were transplanted into 172 immune-deficient SCID mice. Thirty-five percent of SCID mice injected with CLL cells were positive for the presence of human DNA by Southern blot or PCR analysis. The most frequently involved sites were the spleen, lung, kidney and bone marrow, at levels corresponding from 0.1 to 10 percent human DNA. Thrice-weekly intraperitoneal injections of IL-2, alone or in combination with IL-7, did not increase the level of human cell engraftment. SCID mice developed endogenous thymic lymphomas at an incidence of 10-33 percent, a rate that was not increased by CLL cell transplantation. In vitro, CLL cells were able to proliferate for 9 weeks on human stromal layers supplemented with CM (conditioned media from a culture of the human bladder carcinoma cell line 5637), but failed to thrive on the murine stromal cell line MTE cultured either in CM or autologous serum. FACS analysis revealed that 81 percent of proliferating cells on human stromal layers carried the CD5 cell surface marker, identifying them as CLL cells. Previously EBV-negative CLL cells became EBV-positive after 9 to 12 weeks in culture. The results of this study provide a firm foundation for the development of in vivo and in vitro model systems for the study of human CLL. PMID- 8709648 TI - Sensitivity and applicability of different methods for detection of terminal transferase in leukemia. AB - The sensitivity of terminal deoxynucleotidyl transferase (TdT) assay methods was examined by using a mixture of the TdT-positive lymphoblastic leukemia cell line NALM-18 and the TdT-negative erythroleukemia cell line K-562. The biochemical assay could detect TdT activity in the mixture containing NALM-18 cells at concentrations of more than 10 percent. The immunofluorescent (IF) method could detect positive cells in the mixture containing NALM-18 cells at concentrations of more than 1 percent. Furthermore, an approximately 10(5)-fold increase in sensitivity was obtained by the combination of RT-PCR and subsequent Southern blotting, as compared to biochemical assay. In many leukemia cases the expression of TdT-mRNA corresponded well to that of TdT protein. However, in some patients with leukemia, only TdT-mRNA was detectable by RT-PCR without any expression of TdT protein. A PCR-based technique enables us to detect TdT transcripts at the highest sensitivity, but does not allow the characterization of each positive cell. IF analysis is simple and sensitive, but may sometimes cause nonspecific reactions. All these techniques have some advantages and some faults, therefore, the results obtained from clinical studies using these techniques should be interpreted with caution. PMID- 8709649 TI - Flow cytometric detection of intracellular antigens for immunophenotyping of normal and malignant leukocytes. AB - Intracellular antigens are of major importance for immunophenotyping of normal leukocytes as well as leukemias and malignant lymphomas. Immunofluorescence microscopic evaluation of cytocentrifuge preparations has remained the preferred technique for detection of intracellular antigens for a long time. Recently, flow cytometric detection of intracellular antigens has been improved by the development of new permeabilization/fixation solutions. We compared four commercially available solutions: FACS Brand Lysing Solution (FACS Brand; Becton Dickinson, San Jose, CA, USA), Fix & Perm cell permeabilization kit (Fix & Perm; An der Grub, Vienna, Austria), OptiLyse B lysing solution (OptiLyse B; Immunotech, Marseille, France), and ORTHO PermeaFix(PermeaFix; Ortho Diagnostic Systems, Raritan, NJ, USA). These solutions were evaluated for the complexity and duration of the intracellular staining procedure, the effects on light scatter patterns, and the staining results for the intracellular antigens terminal deoxynucleotidyl transferase (TdT), cytoplasmic CD3 (CyCD3), myeloperoxidase (MPO), and cytoplasmic immunoglobulin light chains (CylgL). The four methods could easily be introduced in our laboratory and had only minor effect on the light scatter patterns of the tested cell samples. Each of the four tested antigens was detectable with at least one of the four methods. Only the Fix & Perm cell permeabilization kit could be used for reliable detection of all four intracellular antigens. In a large series of 450 BM and PB samples containing various percentages of TdT+ cells, the results of flow cytometric TdT staining with FACS Brand Lysing Solution were highly comparable to the results obtained by immunofluorescence microscopy (P = <0.00001). Our comparative study shows that flow cytometric detection of the intracellular antigens TdT, CyCD3, MPO, and CylgL can now reliably be performed on a routine basis. PMID- 8709650 TI - Immunophenotypic features and configuration of immunoglobulin genes in hairy cell leukemia-Japanese variant. AB - Immunophenotypes and Ig gene rearrangements were investigated in 12 patients with a variant form of hairy cell leukemia (HCL) termed HCL-Japanese variant (HCL-J), and in an HCL-J-derived cell line. The leukemic cells of HCL-J characteristically showed the phenotype of CD20+, CD5-, CD10-, CD11c+, CD22+, CD24- and CD25-. Ig light (L) chain was undetected in nine cases, and the remaining four cases expressed kappa chain. Expression of Ig heavy (H) chain was studied in nine cases. In addition to Igkappa+ cases showing expression of predominantly gamma H chain isotype, alpha chain was detected in one case without expression of L chain. Rearranged bands in Ig heavy chain (JH) genes were recognized in all 12 cases tested. Rearranged bands in kappa chain genes and germline configuration in chi chain genes were seen in all three Igkappa+ cases tested. Four of nine cases without expression of L chain had a rearranged chi chain gene. The other three cases had chi chain genes in the germline configuration and rearranged and/or deleted kappa chain genes. In the remaining two cases, no rearrangement in either kappa or chi chain genes was detected. The Ig gene configuration and expression in HCL-J, partially overlapping with those described for immature B cell leukemia, were dissociated from the cytological features and CD20+, membrane CD22+ phenotype characteristic of mature B cells. PMID- 8709651 TI - Direct display of hematopoietic tyrosine kinase receptor expression profiles in KG1 cells by PCR using degenerate primers. AB - Hematopoietic tyrosine kinase receptors (HGF-TKRs or class III TKRs) are essential for the growth and differentiation of hematopoietic cells. In this report we present a novel method that generates expression profiles of these receptors. The method was tested and optimized using the myeloblastic/ promyelocytic cell line KG1. The method involves PCR of cDNA using class III specific degenerate primers and subsequent restriction enzyme digests of the 147 bp amplicons followed by fractionation on denaturing poly-acrylamide gels. This primary fingerprint of KG1 revealed equal expression of c-kit and flt3 and to a lesser extent PDGF-R alpha and c-fms. One residual band of unknown origin was seen and appeared to be the proto-oncogene RET following cloning and sequence analysis. This tyrosine kinase receptor is known to play an important role in neural development. In order to detect less abundantly expressed sequences, a secondary fingerprint was generated by pre-digestion of the receptors present in the primary expression profile and subsequent amplification of the residual band. No other tyrosine kinase receptors were observed in KG1. In conclusion, this method allows direct visualization of expression of the HGF-TKRs and has the potential to detect novel homologous receptors. PMID- 8709652 TI - Response to cyclosporin A and recombinant human erythropoietin in a case of B cell chronic lymphocytic leukemia and pure red cell aplasia. PMID- 8709653 TI - p16 gene analysis in multiple myeloma (MM). PMID- 8709654 TI - Parental reciprocation bias in t(9;22) vs genomic imprinting. PMID- 8709655 TI - Identifying poor-quality hospitals with mortality rates. Often there's more noise than signal. PMID- 8709656 TI - Identifying poor-quality hospitals. Can hospital mortality rates detect quality problems for medical diagnoses? AB - OBJECTIVES: Many groups involved in health care are very interested in using external quality indices, such as risk-adjusted mortality rates, to examine hospital quality. The authors evaluated the feasibility of using mortality rates for medical diagnoses to identify poor-quality hospitals. METHODS: The Monte Carlo simulation model was used to examine whether mortality rates could distinguish 172 average-quality hospitals from 19 poor-quality hospitals (5% versus 25% of deaths being preventable, respectively), using the largest diagnosis-related groups (DRGs) for cardiac, gastrointestinal, cerebrovascular, and pulmonary diseases as well as an aggregate of all medical DRGs. Discharge counts and observed death rates for all 191 Michigan hospitals were obtained from the Michigan Inpatient Database. Positive predictive value (PPV), sensitivity, and area under the receiver operating characteristic curve were calculated for mortality outlier status as an indicator of poor-quality hospitals. Sensitivity analysis was performed under varying assumptions about the time period of evaluation, quality differences between hospitals, and unmeasured variability in hospital casemix. RESULTS: For individual DRG groups, mortality rates were a poor measure of quality, even using the optimistic assumption of perfect casemix adjustment. For acute myocardial infarction, high mortality rate outlier status (using 2 years of data and a 0.05 probability cutoff) had a PPV of only 24%, thus, more than three fourths of those labeled poor-quality hospitals (high mortality rate outliers) actually would have average quality. If we aggregate all medical DRGs and continue to assume very large quality differences and perfect casemix adjustment, the sensitivity for detecting poor-quality hospitals is 35% and PPV is 52%. Even for this extreme case, the PPV is very sensitive to introduction of small amounts of unmeasured casemix differences between hospitals. CONCLUSION: Although they may be useful for some surgical diagnoses, DRG-specific hospital mortality rates probably cannot accurately detect poor quality outliers for medical diagnoses. Even collapsing to all medical DRGs, hospital mortality rates seem unlikely to be accurate predictors of poor quality, and punitive measures based on high mortality rates frequently would penalize good or average hospitals. PMID- 8709657 TI - Changes in health after elective percutaneous coronary revascularization. A comparison of generic and specific measures. AB - OBJECTIVES: This study determines changes in health-related quality of life after elective percutaneous transluminal coronary angioplasty and compares generic and specific measures. METHODS: Changes in health-related quality of life were measured in consecutive, symptomatic patients undergoing elective percutaneous coronary revascularization using the Medical Outcomes Study 36-Item Short-Form Health Survey (SF-36), the Specific Activity Scale (SAS), and the Canadian Cardiovascular Society Classification (CCSC). The patients were interviewed as outpatients before admission and at least 6 months later. RESULTS: There were significant changes in the following SF-36 measures: physical functioning (postscore minus prescore = 19.1 +/- 24.1), role limitations due to physical health problems (40.4 +/- 47.2), bodily pain (19.9 +/- 29.3), vitality (12.9 +/- 25.1), social functioning (20.0 +/- 33.1), role limitations due to emotional health problems (26.7 +/- 49.0), and general mental health (7.1 +/- 21.2). General health perceptions did not change significantly. Internal-consistency reliability coefficients for these measures ranged from 0.73 to 0.91. There also was significant improvement in the CCSC class, but the SAS class did not change significantly. Overall, the SF-36 role-physical scale was the most responsive to changes after elective percutaneous coronary revascularization, followed by the CCSC and the SF-36 physical functioning scale. CONCLUSIONS: Although this study cannot determine the causal role of elective percutaneous coronary revascularization in these changes, it provides support for the usefulness of these measures in future evaluations of this intervention. PMID- 8709658 TI - Evaluation of physician intervention letters. AB - OBJECTIVES: The Omnibus Budget Reconciliation Act of 1990 requires that Medicaid Agencies perform drug utilization review (DUR). The Texas Medicaid Agency, in cooperation with the Texas DUR Board, have chosen to mail intervention letters to physicians with patient profiles that indicate possible inappropriate use of medications. The objective of this study was to assess the effect of intervention letters indicating duplicative anti-ulcer medications. METHODS: Analysis of Medicaid prescription claims produced 335 patient profiles involving concurrent therapy. Physicians for 174 patients were selected randomly to receive an intervention letter, a response form, and a stamped envelope. The remaining patients served as a control group. RESULTS: A 71.2% response rate was obtained. Of these responses, 48.9% agreed with the letter and 19.1% disagreed with the letter. Profiles generated 6 months after the letters were sent indicated that 47.7% of the patients in the experimental group were still on concurrent therapy compared with 64.4% of patients in the control group (P = 0.007). CONCLUSIONS: The high response rate to the letter, the moderately high agreement with the letters, and the statistically significant reduction of duplicative therapy in the experimental group indicate that intervention letters can be an effective way to change prescribing. Future research is needed to assess the effects of educational intervention letters for other drug categories, for other populations, and for longer periods of time; and the effect these changes may have on true patient outcomes. PMID- 8709659 TI - The importance of comorbidities in explaining differences in patient costs. AB - OBJECTIVES: The authors examine to what extent comorbidities contribute to differences in patient hospital costs. METHODS: The medical record data for this study were collected from 15 metropolitan Boston hospital for 4,439 patients admitted mostly in 1985 for one of eight common conditions. Massachusetts hospital discharge abstract data for 1985 and 1993 also were used. Comorbidities were identified from the medical record for the 15-hospital data set and from discharge abstracts for all cases. Stepwise regression models were used to develop comorbidity scores. RESULTS: Across all conditions, the medical record based comorbidity score increased the R2 value from .42 in a model with diagnosis related groups alone to .50. In condition-specific analyses, including the comorbidity score increased the R2 by more than 50% in six of eight conditions, and was more important than several other dimensions of severity in explaining condition-specific costs. When comorbidities were determined from discharge abstract data rather than medical records, only approximately half as much comorbidity was found. Also, there was much less explanatory power: the all condition R2 only went from .42 to .44. However, a comorbidity score developed from statewide hospital discharge abstract data was more useful in explaining variations in charges in the eight condition-specific analyses conducted on patients 65 years and older. CONCLUSIONS: Comorbidities, particularly when determined from the medical record, are important determinants of patient costs. PMID- 8709660 TI - Physician response to patient insurance status in ambulatory care clinical decision-making. Implications for quality of care. AB - OBJECTIVES: Individuals without health insurance in general receive fewer health services and are more likely than insured patients to experience poor outcomes. The main goal of this research was to study whether physicians' clinical recommendations vary for insured and uninsured patients, implying that physicians' choices of care may mediate insurance-related differences in health care use. METHODS: The authors designed clinical scenarios that describe routine decisions encountered by primary care physicians in ambulatory settings. Scenarios were designed to include discretionary, nondiscretionary, preventive, and diagnostic/therapeutic services. Insurance status of patients was indicated as either insured or uninsured for the service under consideration. Scenarios were presented to a nationally representative sample of primary care physicians (n = 1182) as part of the American Medical Association 1992 Socio-economic Monitoring System Survey. Physicians were assigned randomly to receive eight scenarios in which patients were either insured or uninsured. For each scenario, physicians were asked to indicate the percentage of patients for whom they would recommend a given service. RESULTS: After controlling for variables associated with nonresponse, we found that physicians who were presented scenarios with insured patients recommended service for 72% of patients, and physicians who were presented scenarios with uninsured patients recommended the same services for 67% of patients (P < 0.001). Physicians recommended both discretionary services (50% versus 42%; P < 0.001) and nondiscretionary services more often for insured than uninsured patients (93% versus 91%; P < 0.05). CONCLUSIONS: In self-reports, physicians are more likely to recommend services for insured than for uninsured patients, and more so when services are discretionary. This provides evidence that physicians' recommendations may be important mediators of insurance-related variation in the use of health-care services. Higher rates of use among the insured may not always reflect higher quality of care, particularly when the service is discretionary in nature. PMID- 8709661 TI - Systemwide provider performance in a Medicaid program. Profiling the care of patients with chronic illnesses. AB - OBJECTIVES: This study illustrates how claims data can be applied to examine cost and clinical performance of providers in the Medicaid program. METHODS: The authors conducted a cross-sectional analysis of Medicaid beneficiaries in Maryland with diabetes mellitus, hypertension, and asthma treated on an ambulatory basis by hospital-based outpatient departments, physician office-based providers, and community health centers. The study year was July 1987 to June 1988. The authors defined the cost performance (high, medium, or low) of providers in the management of each of the three chronic illnesses, both before and after casemix adjustment, using a classification system based on ambulatory diagnoses (ambulatory care groups). The authors constructed claims-based clinical performance indicators for each of the three conditions. These included the number of patients admitted to acute-care hospitals for any and specific (diabetes mellitus, hypertension, and asthma) causes, the number of patients without a follow-up visit within 30 days of being discharged from the hospital, and the number of patients with consecutive emergency room visits during the study period. RESULTS: The ambulatory care group casemix classification system explained 23%, 33%, and 36% of the variation in total payments for patients with hypertension, diabetes, and asthma, respectively. Without adjustment for casemix, 35% to 50% of providers would be misclassified regarding their cost performance. Forty-one (19.4%) of 211 providers who treated all three illnesses were in the same cost group for all three illnesses and 95 (43%) of 223 providers who treated two of the three illnesses were in the same cost group for both illnesses. Among office-based physicians, for all three chronic illnesses, high-cost providers had more admissions (P < 0.01) for ambulatory care-sensitive conditions than low-cost providers. Among hospital outpatient departments, only high-cost providers of asthma had more admissions (P < 0.05) for asthma than low-cost providers. There was no statistically significant (P > 0.05) difference in the clinical performance indicators between high-cost and low-cost hospital outpatient department providers of primary care for hypertensive and diabetic Medicaid beneficiaries. For the other clinical performance indicators, the results were not consistent across the three illnesses or across the different types of providers. CONCLUSIONS: Without adjustments for casemix, a large number of providers are misclassified regarding to cost performance. In addition, most providers are not equally efficient in managing different chronic illnesses. Provider cost performance is not associated consistently with clinical performance, although severity differences not captured by the casemix adjustment may account for these observations. These measurement methods and relationships between provider performance measures may be useful to state Medicaid programs that seek to contain costs, enhance coordination of care, and improve health. PMID- 8709662 TI - Poststroke rehabilitation in older Americans. The Medicare experience. AB - OBJECTIVES: This study examines the poststroke rehabilitation experience for a 20% sample of Medicare patients age 65 years and older admitted to an acute-care hospital with a stroke diagnosis during the 6-month interval, January 1, 1991 to June 30, 1991. Their Medicare claims data are used for two purposes: to identify current poststroke rehabilitation practice in the US population age 65 years and older, and to evaluate the importance of practice variation within this population. METHODS: Regarding the first objective, the authors develop estimates for many poststroke rehabilitation use and cost parameters that formerly were unmeasured. With respect to the second objective, the authors construct and compare average service use rates across all stroke patients in a census division and across all stroke patients residing in the 30 largest metropolitan statistical areas. RESULTS: The authors' Medicare claims analysis indicates that 73% of stroke survivors received either postacute institutional or ambulatory rehabilitation care during the first 6 months poststroke. The published stroke literature, on the other hand, focuses on the minority of stroke survivors, 16.5% in the Medicare data, who were admitted to an inpatient rehabilitation hospital. Regarding the second study objective, the Medicare analysis provides graphic evidence that poststroke rehabilitation practice varies substantially from one geographic area to another and that practice differences translate into large geographic-related differences in the cost of poststroke rehabilitation. CONCLUSIONS: The authors believe the findings demonstrate a problem with inconsistent poststroke rehabilitation practice. PMID- 8709663 TI - Disenrollment of Medicare cancer patients from health maintenance organizations. AB - OBJECTIVES: There is concern that financial incentives in health maintenance organizations (HMOs) might result in pressures to induce sicker members to disenroll. The authors compared disenrollment rates of Medicare HMO enrollees with cancer with disenrollment rates for cancer-free enrollees, using Medicare enrollment files linked to population-based tumor registry data from the Surveillance, Epidemiology, and End Results (SEER) Program. METHODS: The authors identified all aged Medicare beneficiaries who enrolled in an HMO located in a SEER reporting area during 1985 to 1989. Time to disenrollment was analyzed using a proportional hazards model. RESULTS: Overall, cancer patients were no more likely to disenroll than others. However, persons diagnosed with cancer after enrollment were less likely to disenroll than other persons in Independent Practice Association (IPA) mode HMOs (relative risk [RR] = 0.79). Persons diagnosed with cancer after enrollment in group- and staff-model HMOs were about equally likely to disenroll as other persons (RR = 0.91). Persons diagnosed with cancer less than 18 months before enrollment were at high risk for disenrollment from both IPA and group-/staff-model HMOs (RR = 1.47 and 1.35). There was substantial variation among HMOs in overall disenrollment rates and in RRs for disenrollment by cancer patients. CONCLUSIONS: The low disenrollment rates of patients diagnosed after enrollment do not support the contention that features intrinsic to managed care make HMOs unattractive to the seriously ill. Monitoring of selective disenrollment could be used as a screen for possible access and quality problems. PMID- 8709664 TI - A comparison of US and Canadian family physician attitudes toward their respective health-care systems. AB - OBJECTIVES: To examine how family physicians in the United States and Canada evaluate their respective health-care systems. METHODS: The study design was a cross-sectional survey of 300 randomly selected US family physicians and 300 Canadian family physicians. Outcome measures included: attitudes about professional satisfaction and views on health-care delivery and government involvement in health care. RESULTS: Sixty-one percent of the US sample and 67% of the Canadian sample responded. After performing factor and discriminant analyses, we were able to identify nine characteristics differentiating the two groups of physicians. United States physicians were more likely to be older, male, working in rural areas, conservative in their political thinking, and satisfied with their ability to obtain diagnostic tests. United States physicians viewed the health-care system as being in need of fundamental changes, less likely to advocate a central role for government, and more likely to report that litigation concerns influence their clinical decisions. CONCLUSIONS: This study documents differences between Canadian and US family physicians, and suggests that US family physicians might not easily accept a Canadian-style health-care system. Policy makers implementing health-care system changes should be cognizant of the attitudes of physicians that might determine their acceptance or rejection of such changes. PMID- 8709665 TI - Eliminating language barriers for non-English-speaking patients. AB - OBJECTIVES: More than 31 million persons living in the United States do not speak English, therefore language discordance between the clinician and patient may hinder delivery of cost-effective medical care. A new language service was developed in which interpreters are trained in the skills of simultaneous interpretation commonly used at international conferences. The interpreters are linked from a remote site to headsets worn by the clinician and patient through standard communication wires. The service is called "remote-simultaneous interpretation," to contrast it with a traditional method of an interpreter being physically present at the interview and interpreting consecutively "proximate consecutive interpretation." The aim of this study is to assess in a randomized protocol the quality of communication, interpretation, and level of patient, interpreter, and physician satisfaction with these two language services. METHODS: The first postpartum visit with each of 49 mothers and their new born babies was assigned randomly to proximate-consecutive interpretation (control) or to remote-simultaneous interpretation (experimental). Main outcome measures included (1) the number of physician and mother utterances in the visit, (2) the quality of the interpretation, and (3) physician, interpreter, and mother preferences between the two services. RESULTS: The remote-simultaneous interpreter service averaged 8.3 (10%) more physician utterances (95% confidence interval [CI] 4.3, 12.4) and 9.1 (28%) more mother utterances (95% CI 6.1, 12.1). On average, there were 2.8 (12%) fewer inaccuracies of physician utterances in experimental visits compared with control visits (95% CI -5.9, 0.4) and 3.0 (13%) fewer inaccuracies of mother utterances in experimental visits compared with control visits (95% CI -5.4, -0.6). Mothers and physicians significantly preferred the remote-simultaneous service to proximate-consecutive interpretation service. Interpreters stated that they thought mothers and physicians better understood each other using the remote-simultaneous service, although the interpreters preferred to work with the proximate-consecutive service. CONCLUSIONS: Using remote-simultaneous interpretation to improve the quality of communication in discordant-language encounters promises to enhance delivery of medical care for the millions of non-English-speaking patients in the United States. PMID- 8709666 TI - Effectiveness of automatic diagnostic test result feedback on outpatient laboratory and radiology testing in veterans. A controlled trial. PMID- 8709667 TI - [Study of an outbreak of hysteria after hepatitis B vaccination]. AB - BACKGROUND: The aim of this study was to describe an epidemic outbreak of mass hysteria among adolescents which occurred after the immunisation with vaccine against hepatitis B and to determine the association with social variables related to the students. SUBJECTS AND METHODS: All the available information was evaluated and a questionnaire was designed, which included sociodemographic, clinics and school environment variables. A case-control study was conducted. The data were analyzed and outbreak rate and odds ratio with their respective CI (95%) were calculated. RESULTS: 18 cases (among a set of 74 students of seventh of EGB) were clustered in time (10.30-11 h) and space (class of seventh B). There was a clear association between the cases and the female sex (odds ratio = 5.4; CI 95% 1.3 - 23.8). As for the sociological variables there was a association with increasing household members (odds ratio = 5.3; CI 95% 1.3 - 23.4) and with lower educational levels of the father (odds ratio = 10.6; CI 95% 1.2 - 239.9). Among the affected a more unfavorable opinion of the school was detected. CONCLUSIONS: This outbreak fulfils 8 of the 11 criteria classically admitted as usual characteristics of a mass psychogenic illness episode. The sensorial transmission remains demonstrated supporting the hypothesis of a chain reaction. PMID- 8709668 TI - [Perception of the causes of cancer and its prevention in women of Girona]. AB - BACKGROUND: To analyze the perception of causes and prevention of cancer among women, from Girona, Spain, as well as to assess the knowledge of initial symptoms and participation in cancer screening activities. SUBJECTS AND METHODS: Crossectional survey with the following items: cancer risk factors, initial symptoms, perceived susceptibility, knowledge and practice of screening, beliefs regarding prevention and treatment of cancer, intention to attend screening tests and socio-demographic variables. A stratified random sample of 408 women aged 40 to 70 from two counties in Girona, Spain was selected. RESULTS: More than 80% of women identified tobacco, radioactivity and other environmental factors as causes of cancer, being the alcohol identified by 70% of women. It was clearly observed that a high level of these women (97.5%) considered a lump in the breast as an initial symptom of cancer, whereas only a 6.6% thought it was always due to cancer. With regard to cancer screening, 21.9% of women have ever practiced a mammography, 28.5% a cervical citology with Papanicolaou staining and 53.2% self breast examinations. All these screening activities were inversely associated to age (p < 0.05) and directly to educational level (p < 0.05). CONCLUSION: A high knowledge regarding the causes of cancer with the exception of diet was observed in women from Girona, Spain. Knowledge and participation in screening activities is still low in practice of population cancer screening programmes. PMID- 8709669 TI - [Epidemic outbreak of hysteria]. PMID- 8709670 TI - [Methods of review of resource utilization: limitations]. PMID- 8709671 TI - [From Cairo to Pekin: reflections about a problem involving the whole planet]. PMID- 8709672 TI - [Treatment of cardiac arrhythmias during pregnancy]. PMID- 8709673 TI - [Profitability of hypolipidemic drug treatments]. PMID- 8709674 TI - [Profitability of hypolipidemic drug treatment]. PMID- 8709675 TI - [Application of Bayes theorem to clinical diagnosis]. PMID- 8709676 TI - [Shrinking or collapsed lung: that is the question]. PMID- 8709677 TI - [Factors associated with emergency hospital readmission in digestive and hepatobiliary diseases]. AB - OBJECTIVE: To describe the epidemiological characteristics of emergency readmissions due to digestive and hepatobiliary diseases within the hospitals of the Valencian Health Service (VHS), Spain, and to examine their relationship with defined variables related to patients or to previous periods of hospitalization. PATIENTS AND METHODS: Out of the 212,947 episodes of patient hospitalization included in the Uniform Hospital Discharge Data Set of the VHS, excluding those persons under 16 years of age, those who had died in hospital or were discharged due to being transferred to another hospital and also excluding the two hospitals whose record were of poor quality, 18,075 patients were chosen whose principal diagnosis on first admission was one of a digestive or hepatobiliary disease. After a descriptive analysis the relationship was explored between risk of readmission and the variables of age, gender, hospital funding, whether the environment is a rural or urban one, the hospital itself, the diagnostic group, comorbidity, type of admission, length of stay, presence of surgical procedure, discharge criteria and service using an independent Cox Proportional Hazards analysis for each variable and a second Cox analysis adjusted for selected variables. RESULTS: 16.9% of those patients discharged for digestive disorders are re-admitted within a year. The risk of readmission is associated with age, comorbidity, male sex and chronic diseases and, with respect to the previous episode of hospitalization, it is associated with emergency admission, longer period of hospitalization, non-surgical discharge and admission to certain hospitals. CONCLUSION: The risk of emergency readmission is associated with defined variables relating to patient or previous episodes of hospitalization connected with a more serious condition. The risk adjustment carried out could be used in order to identify high-risk cases which could then be paid special attention in order to delay or prevent readmission and as an indirect indicator for use in monitoring the quality of hospital care. PMID- 8709678 TI - CDP-choline: pharmacological and clinical review. AB - Cytidine 5'-diphosphocholine, CDP-choline or citicoline, is an essential intermediate in the biosynthetic pathway of the structural phospholipids of cell membranes, especially in that of phosphatidylcholine. Upon oral or parenteral administration, CDP-choline releases its two principle components, cytidine and choline. When administered orally, it is absorbed almost completely, and its bioavailability is approximately the same as when administered intravenously. Once absorbed, the cytidine and choline disperse widely throughout the organism, cross the blood-brain barrier and reach the central nervous system (CNS), where they are incorporated into the phospholipid fraction of the membrane and microsomes. CDP-choline activates the biosynthesis of structural phospholipids in the neuronal membranes, increases cerebral metabolism and acts on the levels of various neurotransmitters. Thus, it has been experimentally proven that CDP choline increases noradrenaline and dopamine levels in the CNS. Due to these pharmacological activities, CDP-choline has a neuroprotective effect in situations of hypoxia and ischemia, as well as improved learning and memory performance in animal models of brain aging. Furthermore, it has been demonstrated that CDP-choline restores the activity of mitochondrial ATPase and of membranal Na+/K+ ATPase, inhibits the activation of phospholipase A2 and accelerates the reabsorption of cerebral edema in various experimental models. CDP-choline is a safe drug, as toxicological tests have shown; it has no serious effects on the cholinergic system and it is perfectly tolerated. These pharmacological characteristics, combined with CDP-choline's mechanisms of action, suggest that this drug may be suitable for the treatment of cerebral vascular disease, head trauma of varying severity and cognitive disorders of diverse etiology. In studies carried out on the treatment of patients with head trauma, CDP-choline accelerated the recovery from post-traumatic coma and the recuperation of walking ability, achieved a better final functional result and reduced the hospital stay of these patients, in addition to improving the cognitive and memory disturbances which are observed after a head trauma of lesser severity and which constitute the disorder known as postconcussion syndrome. In the treatment of patients with acute cerebral vascular disease of the ischemic type, CDP-choline accelerated the recovery of consciousness and motor deficit, attaining a better final result and facilitating the rehabilitation of these patients. The other important use for CDP-choline is in the treatment of senile cognitive impairment, which is secondary to degenerative diseases (e.g., Alzheimer's disease) and to chronic cerebral vascular disease. In patients with chronic cerebral ischemia, CDP-choline improves scores on cognitive evaluation scales, while in patients with senile dementia of the Alzheimer's type, it slows the disease's evolution. Beneficial neuroendocrine, neuroimmunomodulatory and neurophysiological effects have been described. CDP choline has also been shown to be effective as co-therapy for Parkinson's disease. No serious side effects have been found in any of the groups of patients treated with CDP-choline, which demonstrates the safety of the treatment. PMID- 8709679 TI - HHV8 and skin cancers in immunosuppressed patients. PMID- 8709680 TI - Warning: games seriously damage health. PMID- 8709681 TI - Delta in the real world. PMID- 8709682 TI - Acute bronchiolitis--a perennial problem. PMID- 8709683 TI - Fever and malaria. PMID- 8709684 TI - Preventing pre-eclampsia. PMID- 8709685 TI - Global paradox. PMID- 8709686 TI - Delta: a randomised double-blind controlled trial comparing combinations of zidovudine plus didanosine or zalcitabine with zidovudine alone in HIV-infected individuals. Delta Coordinating Committee. AB - BACKGROUND: Because the benefits of zidovudine (AZT) in HIV-infected individuals are small and do not last long the Delta trial was designed to test whether combinations of zidovudine with didanosine (ddl) or zalcitabine (ddC) were more effective than AZT alone in extending survival and delaying disease progression. METHODS: The trial was randomised, double blind, and international. 3207 participants were allocated to either AZT (600 mg per day) alone (1055), AZT plus ddl (400 mg per day) (1080), or AZT plus ddC (2.25 mg per day) (1072). Participants either had symptoms of HIV disease (if AIDS, with a CD4 cell count of > 50 x 10(6)/L) or a CD4 count of less than 350 x 10(6)/L; 2124 had not had zidovudine before (Delta 1) and 1083 had for at least 3 months (Delta 2). FINDINGS: Over a median follow-up of 30 months, 699 participants died, and 936 of the 2765 without AIDS at entry developed AIDS or died. In participants who had not had AZT before, both combination regimens had substantial benefits in terms of survival (regardless of disease stage at entry); a relative reduction in mortality of 42%, compared to AZT alone (95% Cl 25% to 55%), for AZT plus ddl and of 32% (95% Cl 22% to 47%) for AZT plus ddC. In participants who had had AZT before, the addition of ddl improved survival (p = 0.05; relative reduction 23% [95% Cl 0% to 41%]) but there was no direct evidence of benefit from the addition of ddC (p = 0.47; relative reduction 9% [95% Cl--17% to 29%]). The overall difference in survival between the treatment groups was significant (p < 0.0001; a relative reduction in mortality, compared to AZT alone, of 33% (95% Cl 20% to 44%) for AZT plus ddl and 21% (95% Cl 6% to 34%) for AZT plus ddC). Benefit in terms of disease progression was seen mainly in participants not previously treated with AZT and overall. There was no unexpected toxicity from the combination treatments. INTERPRETATION: Initiation of treatment with combinations of AZT plus ddl or ddC prolongs life and delays disease progression compared with AZT alone. The addition of ddl to participants already treated with AZT also improves survival, although the benefit appears less. PMID- 8709687 TI - Dexamethasone in bronchiolitis: a randomised controlled trial. AB - BACKGROUND: Although corticosteroids are commonly prescribed in the treatment of bronchiolitis, there is no evidence on the efficacy of these drugs in this disorder. We designed a randomised, double-blind, prospective study to assess the efficacy of dexamethasone in infants with bronchiolitis who require hospital management. METHODS: Infants younger than 12 months who had been admitted to hospital for an initial episode of wheezing, were randomly allocated intramuscular dexamethasone (1 mg/kg daily) or placebo, every 24 h for three doses. We excluded infants who were younger than 4 weeks, who required admission to the intensive care unit, or who had a history of congenital heart disease, mechanical ventilation, or supplemental oxygen use. We assessed infants on admission and every 12 h thereafter--vital signs were taken, severity of accessory muscle use and wheezing were measured by a clinical severity score, and pulse oximetry in room air was done. Our primary endpoints were the time to resolution of symptoms--defined as the number of assessments needed to reach oxygen saturation of more than 95% while receiving no supplemental oxygen, an accessory muscle score of 0, a wheeze score of 0 or 1, and resumption of normal feeding--and duration of oxygen therapy. Follow-up assessments were made 10-14 days after discharge by telephone. We used a proportional-hazards model for our survival analysis. FINDINGS: 197 infants presented with bronchiolitis that required inpatient management. 75 were not enrolled (31 no consent, 28 no approach made, 16 transferred elsewhere). Of the 122 enrolled, four were excluded (clinical deterioration, diagnosis of cystic fibrosis, previous intubation, did not receive all study treatment). There were no differences between the dexamethasone (n = 65) and placebo-treated infants in demographic factors, exposure to tobacco smoke, duration of illness, presence of respiratory syncytial virus (RSV) antigen, respiratory rate, or severity score. More dexamethasone treated patients had an initial oxygen saturation of 95% or less (51 [79%] dexamethasone vs 31 [59%] placebo, p = 0.02). There were no differences in duration of oxygen therapy (p = 0.74) or time to resolution of symptoms (p = 0.22). Stratification for presence of RSV antigen or family history of atopy did not affect the results. INTERPRETATION: Our findings do not support the use of dexamethasone in the treatment of bronchiolitis in infants. PMID- 8709688 TI - Randomised controlled comparison of single-dose ciprofloxacin and doxycycline for cholera caused by Vibrio cholerae 01 or 0139. AB - BACKGROUND: Effective antimicrobial therapy can reduce the duration and volume of cholera diarrhoea by half. However, such treatment is currently limited by Vibrio cholerae resistance to the drugs commonly prescribed for cholera, and by the difficulties involved in the administration of multi-drug doses under field conditions. Because of its favourable pharmacokinetics we thought it likely that single-dose ciprofloxacin would be effective in the treatment of cholera. METHODS: In this double-blind study treatment was either a single 1 g oral dose of ciprofloxacin plus doxycycline placebo, or a single 300 mg oral dose of doxycycline plus ciprofloxacine placebo. 130 moderately or severely dehydrated men infected with V cholerae 01 and 130 infected with V cholerae 0139 were randomly assigned treatment. Patients stayed in hospital for 5 days. We measured fluid intake and stool volume every 6 h, and a sample of stool for culture was obtained daily. The primary outcome measures were clinical success--the cessation of watery stool within 48 h; and bacteriological success--absence of V cholerae from cultures of stool after study day 2. FINDINGS: Among patients infected with V cholerae 01, treatment was clinically successful in 62 (94%) of 66 patients who received ciprofloxacin and in 47 (73%) of 64 who receive doxycycline (difference 21% [95% Cl 8-33]); the corresponding proportions with bacteriological success were 63 (95%) and 44 (69%) (27% [14-39]). Among patients infected with V cholerae 0139, treatment was clinically successful in 54 (92%) of 59 patients who received ciprofloxacin and in 65 (92%) of 71 who received doxycycline (< 1% [-9 to 9]), and bacteriologically successful in 58 (98%) and 56 (79%), respectively (19% [9 30]). Total volume of watery stool did not differ significantly between ciprofloxacin-group and doxycycline-group patients infected with either V cholerae 01 or 0139. All but one of the V cholerae 01 and all of the 0139 isolates were susceptible in vitro to doxycycline, whereas 48 (37%) of the V cholerae 01 isolates and none of the 0139 isolates were resistant to tetracycline. Treatment clinically failed in 14 (52%) of 27 doxycycline-treated patients infected with a tetracycline-resistant V cholerae 01 strain, compared with three (8%) of 37 patients infected with a tetracycline-susceptible strain (44% [23-65]). INTERPRETATION: Single-dose ciprofloxacin is effective in the treatment of cholera caused by V cholerae 01 or 0139 and is better than single dose doxycycline in the eradication of V cholerae from stool. Single-dose ciprofloxacin may also be the preferred treatment in areas where tetracycline resistant V cholerae are common. In V cholerae, in-vitro doxycycline susceptibilities are not a useful indicator of the in-vivo efficacy of the drug. PMID- 8709689 TI - Removing bee stings. AB - BACKGROUND: Conventional advice on immediate treatment of honey-bee stings has emphasised that the sting should be scraped off, never pinched. The morphology of the sting suggested little basis for this advice, which is likely to slow down removal of the sting. METHODS: The response to honey-bee stings was assayed with a measurement of the size of the resulting weal. Injection of known quantities of venom showed that this measurement is a good indicator of envenomisation. FINDINGS: Weal size, and thus envenomisation, increased as the time from stinging to removal of the sting increased, even within a few seconds. There was no difference in response between stings scraped or pinched off after 2 s. INTERPRETATION: These data suggest that advice to patients on the immediate treatment of bee stings should emphasise quick removal, without concern for the method of removal. PMID- 8709690 TI - An inhibitor of the sodium pump obtained from human placenta. AB - BACKGROUND: Much effort has been expended in the search for an endogenous inhibitor of the cellular sodium/potassium pump, a compound of major physiological importance, which has been implicated in the mechanism of essential hypertension. Others have suggested that ouabain or an isomer of ouabain may be the endogenous pump inhibitor. Neonatal cord serum contains an inhibitor of the sodium pump; we attempted to isolate and characterise this substance from human placentas. METHODS: Homogenised placentas were dialysed and the resulting solutes were trapped on octadecylsilyl silica and then separated by high-performance liquid chromatography. Measurement of the activity of the sodium pump of human leucocytes was used to test each fraction for the presence of the inhibitor. FINDINGS: An inhibitor of the sodium pump was obtained by this technique in a mass spectrometrically pure form with a mass of 370 Da, an empirical formula of C24H34O3 and only one hydroxyl group. The characteristic fragmentation pattern observed in negative-ion mass spectrometry was compared with those of various model compounds; this comparison suggested that the active material was a dihydropyrone-substituted steroid. INTERPRETATION: These results suggest that a dihydropyrone-substituted steroid is an endogenous regulator of the sodium pump in humans and, presumably, other mammals. Proof of the endogenous origin will require the demonstration of a previously unrecognised biosynthetic pathway. PMID- 8709691 TI - A school-girl with constipation. PMID- 8709692 TI - HIV-associated respiratory diseases. AB - The lungs of individuals infected with HIV are often affected by opportunistic infections and tumours; over two-thirds of patients have at least one respiratory episode during the course of their disease. Despite the availability of effective prophylaxis, infection with the fungus Pneumocystis carinii remains a common cause of respiratory disease. Bacterial infections, which occur more frequently in HIV-infected persons than in the general population, and tuberculosis are increasing causes of morbidity and mortality. Kaposi's sarcoma, the commonest HIV associated malignancy, may affect the lungs in addition to the skin. Pulmonary involvement by non-Hodgkin lymphoma is common in those with disseminated disease. PMID- 8709693 TI - Benefits from specialised cancer care. AB - The Expert Advisory Group's recommendations to establish cancer centres, cancer units, and a network involving primary care for the treatment of cancer patients in the UK is based on the view that specialisation in cancer care will improve outcomes. Evidence to support the various aspects of specialisation (eg, training, caseload, and the formation of multidisciplinary teams) is strongest for breast cancer, ovarian cancer, and some haematological malignant diseases. There is evidence to support the view that some specialised care can be successfully delivered by a network of district hospitals and main general or teaching hospitals and does not always require referral to cancer centres. PMID- 8709694 TI - Fibre-supplemented foods may damage your health. PMID- 8709695 TI - Montreal needle-exchange surprise. PMID- 8709696 TI - Keeping an open mind about prions. PMID- 8709697 TI - US specialists object to AMA's call for mandatory HIV testing. PMID- 8709698 TI - Premenopausal breast cancer after in-utero exposure to stilboestrol. PMID- 8709699 TI - What is genetic anticipation? PMID- 8709700 TI - Infertility in intracytoplasmic-sperm-injection-derived sons. PMID- 8709701 TI - Recombinant human luteinising hormone to mimic mid-cycle LH surge. PMID- 8709702 TI - Non-detection of human papillomavirus DNA in cervical dysplasia and disease progression. PMID- 8709703 TI - APOE genotyping in differential diagnosis of Alzheimer's disease. PMID- 8709704 TI - Apolipoprotein E epsilon 4 in bulbar-onset motor neuron disease. PMID- 8709705 TI - Water treatment and diarrhoea. PMID- 8709706 TI - Manic-depressive illness and tyrosine hydroxylase gene. PMID- 8709707 TI - Riluzole and amyotrophic lateral sclerosis. PMID- 8709708 TI - Reform of the World Health Organization. PMID- 8709709 TI - The measurement of caring. PMID- 8709710 TI - Remak and the plantar response. PMID- 8709711 TI - Driving and the implantable cardioverter defibrillator. PMID- 8709712 TI - The truth of torture cannot be safely told. PMID- 8709713 TI - Fatal overdose with citalopram. PMID- 8709714 TI - Vaccines for Japanese encephalitis. PMID- 8709715 TI - Spiral CT scanning angiography for assessing systemic-to-pulmonary shunt in children. PMID- 8709716 TI - Vaccines for Japanese encephalitis. PMID- 8709717 TI - Total spondylectomy and spinal shortening for giant-cell tumour of spine. PMID- 8709718 TI - Tethered cord syndrome as cause of spinal cord dysfunction. PMID- 8709719 TI - Influence of misleading publications on general practitioners and purchasers. PMID- 8709720 TI - Malaria prophylaxis and the media. PMID- 8709721 TI - Impact of UK malaria prophylaxis policy on imported malaria. PMID- 8709722 TI - Subcutaneous clodronate. PMID- 8709723 TI - Graft-versus-myeloma effect. PMID- 8709724 TI - Torticollis in ancient Macedonians. PMID- 8709725 TI - Buying sex, safely. PMID- 8709726 TI - Stress-related growth failure. PMID- 8709727 TI - The naming of cats--and alpha-interferons. PMID- 8709728 TI - Prophylactic cranial irradiation for small-cell lung cancer. PMID- 8709729 TI - Treatment of symptom-free carriers in management of tinea capitis. PMID- 8709730 TI - Which patients may be harmed by good treatments? PMID- 8709731 TI - Amorous inclinations. PMID- 8709732 TI - A new stress-related syndrome of growth failure and hyperphagia in children, associated with reversibility of growth-hormone insufficiency. AB - BACKGROUND: Growth failure without organic aetiology but associated with behavioural disturbance and psychosocial stress has been termed psychosocial short stature. This condition is not a valid diagnostic entity, but encompasses failure to thrive, stunting secondary to chronic malnutrition, and idiopathic hypopituitarism. Some children show spontaneous catch-up growth when removed from the source of stress, without further treatment, but until now precise definition of this subgroup for the purpose of clinical identification has not been possible. METHODS: Hospital-referred children with growth failure unrelated to organic pathology, who came from stressful homes, were compared with children of short-normal stature identified from an epidemiological survey (n = 31). Growth hormone dynamics were studied in the hospital group by a combination of diurnal profiles and provocation tests. The tests were repeated after a hospital stay of 3 weeks away from familial stress. Standard behavioural measures were obtained from home and school. FINDINGS: In a distinctive subgroup (n = 29), growth hormone insufficiency was associated with characteristic behavioural features, especially hyperphagia and polydipsia, and a normal body-mass index. When the children were removed from their stressful home circumstances, growth-hormone insufficiency spontaneously resolved only in formerly hyperphagic subjects. 74% of the non-hyperphagic cases (n = 23) were anorexic, with a low body-mass index and normal growth-hormone responses to provocation tests. INTERPRETATION: We present explicit behavioural and developmental criteria by which the novel syndrome of hyperphagic short stature may be recognised clinically. Such children have a capacity for spontaneous recovery of growth-hormone production on removal from or reduction of stress. Discriminant and predictive validity of the core symptoms are demonstrated. Preliminary familial studies indicate a possible genetic predisposition. PMID- 8709733 TI - Risk factors for acute myocardial infarction in Indians: a case-control study. AB - BACKGROUND: South Asians who have settled overseas and those in urban India have an increased risk of ischaemic heart disease (IHD). Reasons for this increased risk are unclear. Most studies have been based on migrants to western nations, so their findings may not apply to most south Asians, who live in their own countries. Therefore, we assessed the relative importance of risk factors for IHD among South Asians in Bangalore, India. METHODS: We conducted a prospective hospital-based case-control study of 200 Indian patients with a first acute myocardial infarction (AMI) and 200 age and sex matched controls. We recorded prevalence of the following risk factors for IHD: diet, smoking, alcohol use, socioeconomic status, waist to hip ratio (WHR), blood glucose, serum insulin, oral glucose tolerance test, and lipid profile. FINDINGS: The most important predictor of AMI was current smoking (odds ratio [OR] 3.6, p < 0.001) of cigarettes or beedis (a local form of tobacco), with individuals who currently smoked 10 or more per day having an OR of 6.7 (p < 0.001). History of hypertension and of overt diabetes mellitus were also independent risk factors (OR 2.69 [p = 0.001] and 2.64 [p = 0.004], respectively). Among all individuals, fasting blood glucose was a strong predictor of risk over the entire range, including at values usually regarded as normal (OR adjusted for smoking, hypertension, and WHR 1.62 for 1 SD increase, p < 0.001). Abdominal obesity (as measured by WHR) was also a strong independent predictor across the entire range of measures (OR adjusted for smoking, hypertension, and blood glucose 2.24 for 1 SD increase; p < 0.001). Compared with individuals with no risk factors, individuals with multiple risk factors had greatly increased risk of AMI (eg, OR of 10.6 for the group with smoking and elevated glucose). Lipid profile was not associated with AMI. In univariate analyses, higher socioeconomic (income) status (OR 0.32, p = 0.005 highest vs lowest; OR 0.75 middle vs lowest) and vegetarianism (OR = 0.55, p = 0.006), seemed to be protective. The impact of vegetarianism was closely correlated with blood glucose and WHR. INTERPRETATION: Smoking cessation, treatment of hypertension, and reduction in blood glucose and central obesity (perhaps through dietary modification) may be important in preventing IHD in Asian Indians. PMID- 8709734 TI - Randomised controlled trial of a reduced-visits programme of antenatal care in Harare, Zimbabwe. AB - BACKGROUND: Many of the individual components of antenatal care have been studied in randomised controlled trials, but few studies have compared whole programmes of antenatal care. Our aim was to test the hypothesis that a new programme of antenatal care with fewer goal-oriented visits would give an equivalent or better result in the outcomes associated with pregnancy and delivery. METHODS: In a randomised clinical trial in Harare, Zimbabwe, we compared a new programme of antenatal care with the standard programme. The new programme consisted of fewer but more objectively oriented visits and fewer procedures per visit. Seven primary care clinics were randomly assigned to the two programmes-three to the standard programme and four to the new programme. FINDINGS: Over a 2-year period, 15,994 women were recruited into the study at the time they booked antenatal care. 97% of the women were followed up, 9,394 who had followed the new programme, and 6,138 from clinics with the standard one. Women allocated to the new programme made, as planned, fewer visits than those in the standard programme (median 4 vs 6 visits, respectively). The proportion of antenatal referrals was also lower (13.6 vs 15.3%; odds ratio 0.87 [95% CI 0.79-0.95]) because of significantly fewer referrals for pregnancy-induced hypertension (2.5 vs 3.8%; 0.66 [0.55-0.79]). Nevertheless, there were significantly fewer labour referrals for severe hypertension or eclampsia (2.1 vs 2.6%; 0.81 [0.66-1.00]). The risk for preterm (< 37 weeks) delivery was significantly lower for women on the new programme (10.1 vs 11.5%; 0.86 [0.78-0.96]). There were no other significant differences between the programmes in other major indices of pregnancy outcome, including antenatal referrals for other causes, labour referrals, obstetric interventions, low birthweight, and perinatal and maternal mortality and morbidity. INTERPRETATION: An antenatal care programme with fewer more objectively oriented visits can be introduced without adverse effects on the main intermediate outcome pregnancy variables. PMID- 8709736 TI - Randomised double-blind placebo-controlled study of the effect of inhibition of nitric oxide synthesis in bradykinin-induced asthma. AB - BACKGROUND: Bronchoconstriction induced by bradykinin is reduced by the release of nitric oxide (NO) in the airways of guinea pigs. Inhaled NO is known to cause bronchodilatation in asthmatic patients. To find out the role of endogenous NO in airway response to bradykinin in asthma, we examined the effect of the NO synthase inhibitor NG-monomethyl-L-arginine (L-NMMA) on broncho-constriction after bradykinin challenge in ten patients with mild asthma. METHODS: The study had a randomised, double-blind, placebo-controlled, cross-over design. Participants were studied during two phases, each consisting of 2 study days. After baseline measurements of forced expiratory volume in 1 s (FEV1) participants inhaled an aerosol of L-NMMA or saline (placebo). After 5 min, saline and doubling doses of bradykinin (from 0.25 nmol) were inhaled until FEV1 fell by at least 20% of the post-saline value. The effect of L-NMMA and placebo on airway response to doubling concentrations of methacholine (from 0.03 mg/mL) was then examined. We also assessed the effect of the inactive enantiomer of L NMMA, D-NMMA, and placebo on bronchoconstriction after bradykinin or methacholine challenge in six of the participants. FINDINGS: The geometric mean of the provocative dose producing a 20% fall in FEV1 to bradykinin was 138.0 nmol (range 48.2-475.2 nmol) after placebo and 11.2 nmol (range 0.9-51.3 nmol) after L-NMMA (p < 0.01). L-NMMA also caused a decrease in the provocative concentration of methacholine producing a 20% fall in FEV1 from 0.93 mg/mL (range 0.12-2.55 mg/mL) to 0.38 mg/mL (range 0.06-0.92 mg/mL; p < 0.01). In contrast, D-NMMA did not affect airway response to bradykinin or methacholine. INTERPRETATION: The results suggest that bronchoconstriction after bradykinin inhalation is greatly inhibited by the formation of NO in airways of asthmatic patients and that NO could have a bronchoprotective role in asthma. PMID- 8709735 TI - Clinical evidence of angiogenesis after arterial gene transfer of phVEGF165 in patient with ischaemic limb. AB - BACKGROUND: Preclinical findings suggest that intra-arterial gene transfer of a plasmid which encodes for vascular endothelial growth factor (VEGF) can improve blood supply to the ischaemic limb. We have used the method in a patient. METHODS: Our patient was the eighth in a dose-ranging series. She was aged 71 with an ischaemic right leg. We administered 2,000 micrograms human plasmid phVEGF165 that was applied to the hydrogel polymer coating of an angioplasty balloon. By inflating the balloon, plasmid DNA was transferred to the distal popliteal artery. FINDINGS: Digital subtraction angiography 4 weeks after gene therapy showed an increase in collateral vessels at the knee, mid-tibial, and ankle levels, which persisted at a 12-week view. Intra-arterial doppler-flow studies showed increased resting and maximum flows (by 82% and 72%, respectively). Three spider angiomas developed on the right foot/ankle about a week after gene transfer; one lesion was excised and revealed proliferative endothelium, the other two regressed. The patient developed oedema in her right leg, which was treated successfully. INTERPRETATION: Administration of endothelial cell mitogens promotes angiogenesis in patients with limb ischaemia. PMID- 8709737 TI - A man who coughed for 15 years before a doctor took his pulse. PMID- 8709738 TI - Gastrointestinal manifestations of HIV infection. AB - The harrowing picture of emaciated terminally ill AIDS patients is a reminder of our lack of understanding of immunological mechanisms that normally control opportunistic infections. Many gastrointestinal pathogens in patients with AIDS are resistant to treatment and lead inexorably to weight loss and death. Although knowledge of the pathogenesis and clinical significance of weight loss has improved considerably, this has not yet led to a sustained effort to improve nutritional status during early stages of disease. PMID- 8709739 TI - Systematic review of randomised trials of interventions to assist patients to follow prescriptions for medications. AB - BACKGROUND: Low adherence of patients to prescribed, self-administered medical interventions is ubiquitous. Low adherence limits the benefits of current medical care. Efforts to assist patients to follow treatments might improve the efficiency of care and substantially enhance benefits. Our objective was to summarise the results of randomised controlled trials (RCTs) of interventions to help patients follow prescriptions for medications. METHODS: A previous systematic review was updated through computerised searches in Medline, International Pharmaceutical Abstracts, Psychinfo, and HSTAR online databases; bibliographies in articles on patient adherence; articles in the reviewers' personal collections; and contact with authors. Articles were judged of interest if they reported original data concerning an unconfounded RCT of an intervention to improve adherence with prescribed medications, with one or more measure of medication adherence, one or more measure of treatment outcome, at least 80% follow-up of each group studied, and, for long-term treatments, at least 6 months of follow-up for studies with positive initial findings. Information on study design features, interventions and controls, and findings were extracted by one reviewer (RK) and checked by the other two reviewers. FINDINGS: 1,553 relevant citations and abstracts were screened, 252 full text articles were reviewed in detail, and 13 RCTs met all criteria. The studies were too disparate in clinical problems, adherence interventions, measures and reporting of adherence, and the clinical outcomes studied to warrant meta-analysis. Seven of 15 interventions were associated with improvements in adherence and six interventions led to improvements in treatment outcomes. For short-term treatments, one study showed an effect on adherence and outcome of counselling and written information. The interventions that were effective for long-term care were complex, including various combinations of more convenient care, information, counselling, reminders, self-monitoring, reinforcement, family therapy, and other forms of additional supervision or attention. Even the most effective interventions did not lead to substantial improvements in adherence. INTERPRETATION: Although adherence and treatment outcomes can be improved by certain-usually complex interventions, full benefits of medications cannot be realised at currently achievable levels of adherence. It is time that additional efforts be directed towards developing and testing innovative approaches to assist patients to follow treatment prescriptions. PMID- 8709740 TI - Targeting lipid-lowering drug therapy for primary prevention of coronary disease: an updated Sheffield table. PMID- 8709741 TI - Active tuberculosis boosts HIV replication. PMID- 8709742 TI - Maternal transmission of BSE demonstrated in cattle. PMID- 8709743 TI - Getting under the skin with melanoma vaccines. PMID- 8709744 TI - Mortality from venous thromboembolism and myocardial infarction in young women in the Netherlands. PMID- 8709745 TI - Mortality from venous thromboembolism and myocardial infarction in young adults in England and Wales. PMID- 8709746 TI - Pregnancy-associated osteoporosis. PMID- 8709747 TI - Cholesterol crystal embolism. PMID- 8709748 TI - Desmosterolosis: a new inborn error of cholesterol biosynthesis. PMID- 8709749 TI - Cimetidine and immunoreactivity. PMID- 8709750 TI - Erythrodermic diffuse cutaneous leishmaniasis with Sezary syndrome. PMID- 8709751 TI - Experimentally induced hypoglycaemia. PMID- 8709752 TI - Experimentally induced hypoglycaemia. PMID- 8709753 TI - Association between Fc epsilon RI beta and atopic disorder in Japanese population? PMID- 8709754 TI - Breast cancer studies in The Netherlands. PMID- 8709755 TI - Coproxamol revisited. PMID- 8709756 TI - In-situ breast cancer and BRCA1. PMID- 8709757 TI - International programme on chemical safety. PMID- 8709759 TI - Epidemiology versus a smoke screen. PMID- 8709758 TI - Change in sex ratio with exposure to dioxin. PMID- 8709760 TI - Sponsorship of research. PMID- 8709761 TI - Medical students on the move. PMID- 8709762 TI - Pain in the neck. PMID- 8709763 TI - A new spotted-fever-group rickettsiosis. PMID- 8709764 TI - HIV epidemic in India. PMID- 8709765 TI - Rubella immunisation of men: advantages of herd immunity. PMID- 8709766 TI - Screening tests and diagnosis of dementia. PMID- 8709767 TI - Critical illness polyneuropathy. PMID- 8709768 TI - Critical illness polyneuropathy. PMID- 8709769 TI - Presenilin-1 polymorphism and Alzheimer's disease. PMID- 8709770 TI - Measles case fatality by sex, vaccination status, and HIV-1 antibody in Zambian children. PMID- 8709771 TI - Jet leg, pulmonary embolism, and hypoxia. PMID- 8709772 TI - Erythropoietin production during flights with pressurised aircrafts. PMID- 8709773 TI - Jet leg, pulmonary embolism, and hypoxia. PMID- 8709774 TI - Life on Mars is bigger than all of us. PMID- 8709775 TI - Endothelin-1 in heart failure: a new therapeutic target? PMID- 8709776 TI - Congenital defects, folic-acid, and homoeobox genes. PMID- 8709777 TI - Protection against Alzheimer's disease? PMID- 8709778 TI - Quality of quality-of-life data. PMID- 8709779 TI - Adenomatous polyposis coli and translational medicine. PMID- 8709780 TI - Bleeding complications of oral anticoagulant treatment: an inception-cohort, prospective collaborative study (ISCOAT). Italian Study on Complications of Oral Anticoagulant Therapy. AB - BACKGROUND: Bleeding is the most serious complication of the use of oral anticoagulation in the prevention and treatment of thromoboembolic complications. We studied the frequency of bleeding complications in outpatients treated routinely in anticoagulation clinics. METHODS: In a prospective cohort from thirty-four Italian anticoagulation clinics, 2745 consecutive patients were studied from the start of their oral anticoagulation (warfarin in 64%, acenocourmarol in the rest). The target anticoagulation-intensity was low (international normalised ratio [INR] < or = 2.8) in 71% of the patients and high (> 2.8) in the remainder. We recorded demographic details and the main indication for treatment and, every 3-4 months, INR and outcome events. Such events included all complications (bleeding, thrombosis, other), although only bleeding events are reported here, and deaths. We divided bleeding into major and minor categories. FINDINGS: 43% of the patients were women. Nearly three-fifths of the patients were aged 60-79; 8% were over 80. The main indication for treatment was venous thrombolism (33%), followed by non-ischaemic heart disease (17%). Mean follow-up was 267 days. Over 2011 patient-years of follow-up, 153 bleeding complications occurred (7.6 per 100 patient-years). 5 were fatal (all cerebral haemorrhages, 0.25 per 100 patient-years), 23 were major (1.1), and 125 were minor (6.2). The rate of events was similar between sexes, coumarin type, size of enrolling centre, and target INR. The rate was higher in older patients: 10.5 per 100 patient-years in those aged 70 or over, 6.0 in those aged under 70 (relative risk 1.75, 95% Cl 1.29-2.39, p < 0.001). The rate was also higher when the indication was peripheral and/or cerebrovascular disease than venous thromboembolism plus other indications (12.5 vs 6.0 per 100 patient-years) (1.80, 1.2-2.7, p < 0.01), and during the first 90 days of treatment compared with later (11.0 vs 6.3, 1.75, 1.27-2.44, p < 0.001). A fifth of the bleeding events occurred at low anticoagulation intensity (INR < 2, rate 7.7 per 100 patient years of follow-up). The rates were 4.8, 9.5, 40.5, and 200 at INRs 2.0-2.9, 3 4.4, 4.5-6.9, and over 7, respectively (relative risks for INR > 4.5, 7.91, 5.44 11.5, p < 0.0001). INTERPRETATION: We saw fewer bleeding events than those recorded in other observational and experimental studies. Oral anticoagulation has become safer in recent years, especially if monitored in anticoagulation clinics. Caution is required in elderly patients and anticoagulation intensity should be closely monitored to reduce periods of overdosing. PMID- 8709781 TI - Effect of oestrogen during menopause on risk and age at onset of Alzheimer's disease. AB - BACKGROUND: Oestrogen use by postmenopausal women has many health benefits, but findings on the effect of oestrogen in Alzheimer's disease are conflicting. Oestrogen promotes the growth and survival of cholinergic neurons and could decrease cerebral amyloid deposition, both of which may delay the onset or prevent Alzheimer's disease. To investigate whether use of oestrogen during the postmenopausal period affects the risk of Alzheimer's disease, we studied 1124 elderly women who were initially free of Alzheimer's disease, Parkinson's disease, and stroke, and who were taking part in a longitudinal study of ageing and health in a New York City community. METHODS: Relative risks and age-at-onset distributions were calculated from simple and adjusted Cox proportional hazards models. Standard annual clinical assessments and criterion-based diagnoses were used in follow-up (range 1-5 years). FINDINGS: Overall, 156 (12.5%) women reported taking oestrogen after onset of menopause. The age at onset of Alzheimer's disease was significantly later in women who had taken oestrogen than in those who did not and the relative risk of the disease was significantly reduced (9/156 [5.8%] oestrogen users vs 158/968 [16.3%] nonusers; 0.40 [95% Cl 0.22-0.85], p < 0.01), even after adjustment for differences in education, ethnic origin, and apolipoprotein-E genotype. Women who had used oestrogen for longer than 1 year had a greater reduction in risk; none of 23 women who were taking oestrogen at study enrolment has developed Alzheimer's disease. INTERPRETATION: Oestrogen use in postmenopausal women may delay the onset and decrease the risk of Alzheimer's disease. Prospective studies are needed to establish the dose and duration of oestrogen required to provide this benefit and to assess its safety in elderly postmenopausal women. PMID- 8709782 TI - Molecular genetic tests as a guide to surgical management of familial adenomatous polyposis. AB - BACKGROUND: In familial adenomatous polyposis the only curative treatment is colectomy, and the choice of operation lies between restorative proctocolectomy (RPC) and colectomy with ileorectal anastomosis (IRA). The RPC procedure carries a higher morbidity but, unlike IRA, removes the risk of subsequent rectal cancer. Since the course of familial adenomatous polyposis is influenced by the site of mutation in the polyposis gene, DNA analysis might be helpful in treatment decisions. METHODS: We evaluated the incidence of rectal cancer in polyposis patients who had undergone IRA, and examined whether the requirement for subsequent rectal excision because of cancer or uncontrollable polyps was related to the site of mutation. FINDINGS: Between 1956 and mid-1995, 225 patients registered at the Netherlands Polyposis Registry had undergone IRA. In 87 of them, a pathogenetic mutation was detected. 72 patients had a mutation located before codon 1250 and 15 patients after this codon. The cumulative risk of rectal cancer 20 years after surgery was 12%, and at that time 42% had undergone rectal excision. The risk of secondary surgery was higher in patients with mutations in the region after codon 1250 than in patients with mutations before this codon (relative risk 2.7, p < 0.05). INTERPRETATION: On this evidence, IRA should be the primary treatment for polyposis in patients with mutations before codon 1250, and RPC in those with mutations after this codon. PMID- 8709783 TI - Re-emergence of epidemic Venezuelan equine encephalomyelitis in South America. VEE Study Group. AB - BACKGROUND: Venezuelan equine encephalomyelitis (VEE) virus has caused periodic epidemics among human beings and equines in Latin America from the 1920s to the early 1970s. The first major outbreak since 1973 occurred in Venezuela and Colombia during 1995, and involved an estimated 75,000 to 100,000 people. We report an epidemiological and virological investigation of this epidemic. METHODS: Virus isolates were made in cell culture from human serum, human throat swabs, and brain tissue from aborted and stillborn human fetuses, as well as from horse brain tissue and pooled mosquito collections. Human sera were also tested for VEE-specific antibodies. The serotypes of VEE isolates were identified by antigen assays, and viruses were characterised genetically by sequencing PCR products generated from the E3 and E2 genes. Phylogenetic analyses were done to determine evolutionary relations with respect to previous epidemic/epizootic and enzootic VEE virus isolates. Mosquito collections were made to identify possible vectors, and clinical findings were determined by direct observation of patients visiting hospitals and clinics in affected regions, and by inspecting patient records. Equine vaccination and vector control were used in an attempt to halt the spread of the outbreak. FINDINGS: Most affected people had an acute, self limited febrile illness of 3 to 4 days duration. However, convulsions were often seen in children, and abortions and fetal deaths occurred in pregnant women infected with VEE virus. Antigenic characterisation of 12 virus isolates spanning the temporal and spatial range of the outbreak indicated that all are VEE serotype IC. Phylogenetic analysis revealed that all of the 1995 viruses were closely related to serotype IC viruses isolated during a large VEE outbreak that occurred in the same regions of Colombia and Venezuela from 1962-1964. A 1983 mosquito isolate from north central Venezuela was also closely related to the 1995 isolates. INTERPRETATION: This outbreak was remarkably similar to one that occurred in same regions of Venezuela and Colombia during 1962-1964. Symptoms of infected patients, estimated mortality rates, meteorological conditions preceding the epidemic, and seasonal patterns of transmission were all very similar to those reported in the previous outbreak. In addition, viruses isolated during 1995 were antigenically and genetically nearly identifical to those obtained during 1962-1964. These findings suggest that the epidemic resulted from the re emergence of an epizootic serotype IC VEE virus. Identification of a similar virus isolate in mosquitoes in Venezuela in 1983, 10 years after epidemic/epizootic VEE activity ceased, raises the possibility of a serotype IC enzootic transmission cycle in northern Venezuela. PMID- 8709784 TI - Nosocomial infection with Rhizopus microsporus in preterm infants: association with wooden tongue depressors. AB - BACKGROUND: We report four cases of cutaneous infection with Rhizopus microsporus in vulnerable preterm infants in one neonatal nursery. METHODS: There was no overlap in hospital stay between the first two cases and an extensive programme of environmental sampling was therefore initiated, with special attention being paid to items that had close contact with infants' skin. Samples were cultured by standard techniques, including fungal culture with Sabouraud's medium. Filamentous fungi with non-septate hyphae were presumptively identified as agents of mucormycosis and referred to the Mycology Reference Laboratory, Bristol, UK, for speciation. FINDINGS: The source of infection was identified as wooden tongue depressors, which were used on the nursery to construct splints for intravenous and arterial cannulation sites. The outbreak was ended by the removal of these items from the nursery. INTERPRETATION: Wooden tongue depressors can be a vehicle for transmission of mucormycosis. The combination of warm, humid conditions in neonatal incubators, particularly in association with occlusive dressings, may favour cutaneous fungal invasion and put small, sick babies at risk of infection. PMID- 8709785 TI - A porter with pain in his neck. PMID- 8709786 TI - Neurological complications of HIV infection. AB - Neurological complications of HIV infection cause considerable morbidity and are often associated with high mortality. These complications include not only the more common opportunistic diseases affecting the brain (cerebral toxoplasmosis, primary central nervous system lymphoma, progressive multifocal leucoencephalopathy, and cryptococcal meningitis) but also the AIDS dementia complex, with its characteristic cognitive and motor dysfunction, which is caused by HIV itself. Additionally, the peripheral nervous system is the target of several disorders, including a common painful neuropathy. Because these and other, less common, central and peripheral nervous system complications of HIV can often be specifically treated or effectively palliated, their accurate and timely diagnosis is important. PMID- 8709787 TI - How should cadaver kidneys be allocated? PMID- 8709788 TI - The politics of grafting cadaver kidneys. PMID- 8709789 TI - Cadaver kidneys: the rules of rationing. PMID- 8709790 TI - Images of physicians in literature: from quacks to heroes. PMID- 8709791 TI - HIV-1-resistant individuals may lack HIV-1 coreceptor. PMID- 8709792 TI - Bio-engineered tissues move towards the clinic. PMID- 8709793 TI - Multiresistant Salmonella typhimurium DT104 in cats: a public health risk. PMID- 8709794 TI - Aplastic anaemia after HGV infection. PMID- 8709795 TI - Hepatitis G-associated aplastic anaemia. PMID- 8709796 TI - Fludarabine treatment and transfusion-associated graft-versus-host disease. PMID- 8709797 TI - Fludarabine treatment and transfusion-associated graft-versus-host disease. PMID- 8709798 TI - Childhood cancer mortality. PMID- 8709799 TI - Childhood cancer mortality. PMID- 8709800 TI - European strategies in the war on cancer. PMID- 8709801 TI - 'Chicane-like' airway as a complication of lingual tonsils. PMID- 8709802 TI - HHV-8 in lymphoproliferative lesions in skin. PMID- 8709803 TI - Chernobyl and hypothyroidism. PMID- 8709804 TI - Exercise, wellbeing, and endogenous molecules of mood. PMID- 8709805 TI - Exercise, wellbeing, and endogenous molecules of mood. PMID- 8709806 TI - Jewish women, breast cancer, and commercial risks. PMID- 8709807 TI - Abortion and fertility regulation. PMID- 8709808 TI - Abortion and fertility regulation. PMID- 8709809 TI - Abortion and fertility regulation. PMID- 8709810 TI - Croatia's Academy of Medical Science. PMID- 8709811 TI - Poor recall performance of journal-browsing doctors. PMID- 8709812 TI - Outpatients care of HIV-1-infected destitute people: a study in Paris, France. PMID- 8709813 TI - Global burden of the HIV epidemic. PMID- 8709814 TI - Biology of tapeworm disease. PMID- 8709815 TI - Biology of tapeworm disease. PMID- 8709816 TI - Transient urinary incontinence after botulinum A toxin. PMID- 8709817 TI - Effect of frequent prenatal ultrasound on birthweight: follow up at 1 year of age. Western Australian Pregnancy Cohort (Raine) Working Group. PMID- 8709818 TI - Apolipoprotein-E genotyping in diagnosis of Alzheimer's disease. PMID- 8709819 TI - Apolipoprotein-E genotyping in diagnosis of Alzheimer's disease. PMID- 8709820 TI - Diagnostic pitfalls and treatment in a young man with systemic amyloidosis. PMID- 8709821 TI - Are aspirin and other non-steroidal anti-inflammatory drugs effective in the prevention and treatment of colorectal cancer? PMID- 8709822 TI - Platelet glycoprotein IIIa PlA polymorphism in young men with myocardial infarction. PMID- 8709823 TI - Diphtheria in eastern Europe. PMID- 8709824 TI - Pill scares and public responsibility. PMID- 8709825 TI - [Between extremes--plea for differentiated surgical therapy of intra-abdominal infections]. PMID- 8709826 TI - [Photodynamic therapy of dysplasia and early cancer of the esophagus]. AB - Photodynamic therapy (PDT) is a new local, endoscopically controlled therapeutic concept based on the selective sensitization of malignant and precancerous lesions prior to light-induced tissue destruction. The first therapeutic experiences with the conventional photosensitizer dihematopor-phyrineter in the treatment of Barrett's esophagus without or with severe dysplasia and mucosal carcinomas of the esophagus demonstrate the efficacy and the curative potential of photodynamic therapy. The unwanted skin photosensitivity and only relative selectivity of the haematoporphyrins however, are somehow unsatisfactory and therefore, an optimization of the method of PDT is necessary. The endogenous photosensitizer aminolaevulinic acid with its high mucosa specificity without phototoxic side effects of the skin which can be administered orally was successfully used by our group for the treatment of patients with severe dysplasia of Barrett's esophagus and superficial squamous cell carcinomas. In patients with severe dysplasia/carcinoma in situ of Barrett's esophagus and early squamous cell carcinomas who pose high surgical risk, photodynamic therapy might be an effective alternative treatment modality in the future. PMID- 8709827 TI - [Prevention of colorectal carcinoma. Current WHO guidelines for early detection of colorectal carcinoma. World Health Organization Collaborating Center for the Prevention of Colorectal Cancer]. PMID- 8709828 TI - [Differentiated surgical therapy of diffuse peritonitis--a prospective study]. AB - PATIENTS AND METHODS: In a prospective protocol 86 consecutive patients with a diffuse peritonitis were treated in the Surgical Clinic of the RWT-University in Aachen (FRG) from January 1992 to June, 1994. According to the "Mannheim Peritonitis-Score" three different stages were treated with different procedures. Stage-I-peritonitis (mild forms, MPS 0-20) was treated with the so-called standard procedure, Stage-II-cases (MPS: 21-29) with the closed postoperative lavage and severe stage-III-cases (MPS > 29) with the so-called "Etappenlavage" (multiple reexplorations and intraoperative lavage). RESULTS: 36 patients showed stage-I-, 29 stage-II-, and 21 stage-III-peritonitis. Mortality was 3% (1/36) in stage-I-, 24% (7/29) in stage-II-, and 33% (7/21) in stage-III-groups. The overall mortality of the whole group was 17% (15/86). The statistically expected mortality was 38% according to the MPS and 33% related to the APACHE-II-Score. PMID- 8709829 TI - [Surgical treatment of intra-abdominal abscesses]. AB - To define the role of operative treatment of intraabdominal abscesses, we retrospectively reviewed 106 patients, who were operated for intraabdominal abscesses between January 1988 and October 1994. 49 patients had had a primary abscess, 57 patients had an anteceding operation. In both groups appendix and bilio-pancreatic tract were the mostly involved organs. 81 (76,4%) abscesses were single, whereas 17 patients had 2 and 8 patients had 3 synchronous abscesses. All abscesses fulfilled the criteria of complicated abscesses. Successful drainage was achieved in 83%, 17% of cases had to be reoperated or percutaneously drained for recurrence of abscess or abscess related complications. The procedure related morbidity was 37,7% (40/106), half of these were minor complications. 5 patients died, conferring to a mortality of 4,7%. We conclude, that operative drainage plays an important role in the therapeutic regimes of intraabdominal complicated abscesses and can be performed with low morbidity and mortality. PMID- 8709830 TI - [Liver cell carcinoma as a late complication of Alagille syndrome (arterio hepatic dysplasia)]. AB - The Alagille syndrome which is also known as arterio-hepatic dysplasia is an autosomal dominant inherited disorder. In several cases cytogenetic studies revealed an interstitial deletion of the short arm of chromosome 20. The hypoplasia or paucity of the interlobular bile ducts causes a chronic intrahepatic cholestasis. The association with facial dysmorphia, embryotoxon posterior, pulmonary stenosis and vertebral deformities are required for the diagnosis of the complete Alagille syndrome. The occurrence of hepatocellular carcinoma as a late complication of the Alagille syndrome was recognized only 11 years after the first publication by Alagille et al. So far 15 cases complicated by hepatocellular carcinoma have been reported. There is one family where all four siblings suffered from hepatocellular carcinoma. Our own case concerns a 31 year old man who died of hepatocellular carcinoma. The postmortem study of his medical history reaching back to childhood allowed the diagnosis of an unrecognized Alagille syndrome. PMID- 8709831 TI - [Individual therapeutic procedure in metastatic carcinoid of the small intestine]. AB - Actually, the therapeutic goal in a metastasized carcinoid tumor is rather symptomatic than curative. Here we report the case of a 53 year old woman with a hepatic and pulmonal metastasized carcinoid tumor of the small intestine. We will discuss current therapeutic options and the approach to the individual patient. After treatment with the standard therapeutics Octreotide and Interferon alpha we could achieve a remarkable and persistent remission by applying the rather unusual drug Dacarbacine in this patient. This is an example for the frequently occurring clinical situation in the treatment of patients with rare tumors requiring the application of individually modified therapeutic regimens. Furthermore, the potency of Dacarbacine in the treatment of carcinoid tumors seems to be underestimated up to now. PMID- 8709832 TI - [Modified triple therapy: what degree of acid inhibition is necessary?]. PMID- 8709833 TI - Discovering asymptomatic biochemical abnormalities on a Baltimore internal medicine service. AB - To assess the frequency of abnormalities that could be discovered through biochemical profile screening on patients admitted to a city hospital internal medicine ward service, we conducted a prospective cross-sectional chart and laboratory review. All unassigned patients admitted to the general medicine service during 1- to 2-month period in late 1993 and the spring of 1994 were eligible. The main outcome measures were frequency of abnormal test results and identification of significance. Admitted patients (N = 222) were evaluated with a 24-panel biochemical profile. Of 5,328 tests, 29% were outside the standard reference range. Of 3,851 tests classified as screening, 1,049 (27%) were outside the reference range. Of overall screening tests, 741 (19%) were judged potentially important by the predetermined criteria. The prevalence did not differ significantly when analyzed by age, race, gender, or history of substance abuse. Our experience indicates that asymptomatic biochemical abnormalities are common in patients admitted to a city hospital medical service and that admission biochemical screening is an effective method of identifying potential comorbidity. Further studies are needed to assess the impact of this approach. PMID- 8709834 TI - A 72-year-old man with shortness of breath. PMID- 8709835 TI - Treatment of depression in the elderly. PMID- 8709836 TI - Imaging case of the month. Distal small bowel obstruction caused by lymphoma. PMID- 8709837 TI - Focused professional education: a response to physicians' individual educational needs. PMID- 8709838 TI - Molecular handles on adaptive mutation. AB - In one experimental system, several handles on the molecular mechanism of apparent adaptive mutation have emerged. The system is reversion of a lac frameshift mutation in Escherichia coli. The molecular handles include a requirement for homologous recombination; the implication of DNA double-strand breaks as a molecular intermediate; a unique sequence spectrum of -1 deletions in mononucleotide repeats which implies polymerase errors, and also implies a failure of postsynthesis mismatch repair on those errors; and the involvement of sexual functions at some stage of the process. These molecular handles are revealing an unexpected new mechanism of mutagenesis. PMID- 8709839 TI - Transcription antitermination: the lambda paradigm updated. AB - Coliphage lambda employs systems of transcription termination and antitermination to regulate gene expression. Early gene expression is regulated by the phage encoded N protein working with a series of Escherichia coli proteins, Nus, at RNA sites, NUT, to modify RNA polymerase to a termination-resistant form. Expression of lambda late genes is regulated by the phage-encoded Q antitermination protein. Q, which appears to use only one host factor, acts at a DNA site, qut, to modify RNA polymerase to a termination-resistant form. This review focuses on recent studies which show that: (i) N can mediate antitermination in vitro, independent of Nus proteins. (ii) Early genes in another lambdoid phage HK022 are also regulated by antitermination, where only an RNA signal appears necessary and sufficient to create a termination-resistant RNA polymerase. (iii) A part of the qut signal appears to be read from the non-template DNA strand. (iv) A host encoded inhibitor of N antitermination appears to act through the NUT site as well as with the alpha subunit of RNA polymerase, and is antagonized by NusB protein. PMID- 8709840 TI - A role for bacteriophages in the evolution and transfer of bacterial virulence determinants. AB - A virulence-associated region in the genome of Dichelobacter nodosus has been shown to contain an integrase gene which is highly related to the integrases of Shigella flexneri phage Sf6 and coliphages P4 and phi R73, together with open reading frames (vapB, C and D) related to genes borne on plasmids in Neisseria gonorrhoeae, Escherichia coli, Actinobacillus actinomycetemcomitans and Treponema denticola. Similar to P4 and phi R73, the vap region is bracketed by putative bacteriophage att sites and is adjacent to a tRNA gene, which suggests that the vap region has been derived by the integration of a bacteriophage, or a plasmid carrying a bacteriophage-related integrase gene. Many similarities in genes and genes clusters encoding virulence determinants have been found in distantly related bacteria. These genes are often located on plasmids in one organism but on the chromosome in others, implying that transmission of the genes has been followed by integration. Thus, the events which have generated the vap regions of D. nodosus may represent a common mechanism for transfer of virulence determinants. A number of genes involved in the virulence of bacterial pathogens are found on integrated bacteriophages, and we suggest that others will prove to be associated with tRNA genes and/or integrase genes derived from bacteriophages. The use of tRNA genes as integration sites for many bacteriophages and plasmids may favour intergeneric transmission, as tRNA genes are highly conserved. PMID- 8709841 TI - Lipopolysaccharide with an altered O-antigen produced in Escherichia coli K-12 harbouring mutated, cloned Shigella flexneri rfb genes. AB - Cloning of the rfb genes of Shigella flexneri 2a into Escherichia coli K-12 strain DH1 results in the synthesis of lipopolysaccharides (LPS) with an O antigen chain having type antigen IV and group antigens 3,4. During genetic studies of these rfb genes in E. coli K-12, we observed that strains harbouring plasmids with certain mutations (inversion and transposon insertions) which should have blocked O-antigen synthesis nevertheless still produced LPS with O antigen chains. These LPS migrated differently on silver-stained SDS polyacrylamide gels, compared with the LPS produced by wild-type rfb genes, and the group 3,4 antigens were barely detectable, suggesting that the O-antigen was altered. Investigation of the genetic determinants for production of the altered O-antigen/LPS indicated that: (i) these LPS are produced as a result of mutations which are either polar on rfbF or inactivate rfbF; (ii) the rfbX gene product (or a similar protein in the E. coli K-12 rfb region) is needed for production of the altered O-antigen in the form of LPS; (iii) the rfbG gene product is required for the production of both the parental and altered LPS; (iv) the dTDP-rhamnose biosynthesis genes are required. Additionally, an E. coli K-12 gene product(s) encoded outside the rfb region also contributes to production of the O-antigen of the altered LPS. An antiserum raised to the altered LPS from strain DH1(pPM2217 (rfbX::Tn1725)) was found to cross-react with nearly all S. flexneri serotypes, and with the altered LPS produced by other DH1 strains harbouring plasmids with different rfb mutations, as described above. The reactivity of the altered LPS with a panel of monoclonal antibodies specific for various S. flexneri O-antigen type and group antigens demonstrated that their O-antigen components were closely related to that of S. flexneri serotype 4. The RfbF and RfbG proteins were shown to have similarity to rhamnose transferases, and we identified a motif common to the N-termini of 6-deoxy-hexose nucleotide sugar transferases. We propose that the E. coli K-12 strains harbouring the mutated S. flexneri rfb genes produce LPS with a hybrid O-antigen as a consequence of inactivation of RfbF and complementation by an E. coli K-12 gene product. Analysis of the genetic and immunochemical data suggested a possible structure for the O-antigen component of the altered LPS. PMID- 8709842 TI - Induction of complex intracytoplasmic membranes related to nitrogen fixation in Azoarcus sp. BH72. AB - We report the discovery of novel subcellular structures related to bacterial nitrogen fixation in the strictly respiratory diazotrophic bacterium Azoarcus sp. BH72, which was isolated as an endophyte from Kallar grass. Nitrogenase is derepressed under microaerobic conditions at O2 concentrations in the micromolar range. With increasing O2 deprivation, bacteria can develop into a hyperinduced state, which is characterized by high specific rates of respiration and efficient nitrogen fixation at approximately 30 nM O2. Ultrastructural analysis of cells in the course of hyperinduction revealed that complex intracytoplasmic membrane systems are formed, which consist of stacks of membranes and which are absent under standard nitrogen-fixing conditions. The iron protein of nitrogenase was highly enriched on these membranes, as evidenced by immunohistochemical studies. Membrane deficiency in NifH/K- mutants, a deletion mutant in the nifK gene and the character of NH+4-grown cells suggested, in concert with the membrane localization of nitrogenase, that these structures are specialized membranes related to nitrogen fixation. We propose the term 'diazosomes' for them. Development of intracytoplasmic membranes coincides with the appearance of a high molecular-mass form of the iron protein of nitrogenase, which was detectable in membrane fractions. Mutational analysis, and determination of the N-terminal amino acid sequence indicate that the nifH gene product is covalently modified by a mechanism probably different from adenosine diphosphoribosylation. Development of diazosomes in nitrogen-fixing cells can be induced in pure cultures and in co culture with a fungus isolated from the rhizosphere of Kallar grass. PMID- 8709843 TI - TolC and DsbA are needed for the secretion of STB, a heat-stable enterotoxin of Escherichia coli. AB - STB secretion-deficient mutants were isolated using the synthetic transposon Tn beta laM. Cultures were plated using a double-membrane system of cellulose acetate and nitrocellulose placed on Luria agar plates containing carbenicillin. The STB bound to the underlying nitrocellulose membrane was detected with anti STB antibodies. The altered genes of two STB secretion-deficient mutants were identified by conjugation and complementation as tolC and dsbA. In cultures of well-characterized dsbA and tolC mutants, STB was absent from the culture supernatant. The role of TolC and DsbA in the secretion of peptides is discussed. PMID- 8709844 TI - Regulation of the heat-shock response depends on divalent metal ions in an hflB mutant of Escherichia coli. AB - HflB, also called FtsH, is an essential Escherichia coli protein involved in the proteolysis of the heat-shock regulator sigma 32 and of the phage regulator lambda cll. The hflB1(Ts) allele (formerly called ftsH1) conferring temperature sensitive growth at 42 degrees C is suppressed by loss of the ferric-uptake repressor Fur and by anaerobic growth. We show here that suppression requires TonB-dependent Fe(III) transport in the hflB1(Ts) fur mutant during aerobic growth at 42 degrees C and Feo-dependent Fe(II) transport during anaerobic growth at 42 degrees C. Temperature-resistant growth of hflB1(Ts) strains is also observed at 42 degrees C in the presence of a high concentration of Fe(II), Ni(II), Mn(II) or Co(II) salts, but not in the presence of Zn(II), Cd(II), Cu(II), Mg(II), Ca(II) or Cr(III) salts. However, neither Ni(II) nor a fur mutation permits growth in the complete absence of HflB. The heat-shock response, evaluated by an htpG::lacZ fusion, is overinduced in hflB1(Ts) strains at 42 degrees C because of stabilization of sigma 32. Growth in the presence of Ni(II) or in the absence of the Fur repressor abolishes this overinduction in the hflB1(Ts) strain, and, in the hflB1(Ts) fur mutant, sigma 32 is no longer stabilized at 42 degrees C. These results reinforce the recent observation that HflB is a metalloprotease active against sigma 32 in vitro and suggest that it can associate functionally in vivo with Fe(II), Ni(II), Mn(II) and Co(II) ions. PMID- 8709845 TI - Evidence for lateral transfer and recombination in OspC variation in Lyme disease Borrelia. AB - The ospC gene was amplified by the polymerase chain reaction from each of 76 Lyme disease Borrelia strains. Restriction fragment length polymorphism (RFLP) analysis demonstrated 33 distinct RFLP types; two additional RFLP types were identified from published ospC sequences. For each RFLP type, at least one ospC gene was sequenced and the degree of sequence relatedness examined by construction of an ospC gene tree. The genes were extremely diverse, with sequence identity ranging from 74.4% to 99.0%; the majority of changes are localized within the central portion of the molecule. A comparison of ospC sequences suggests that recombination occurs frequently between ospC alleles; this genetic exchange is proposed to be mediated by lateral transfer of ospC sequences. Evidence indicates that recombination occurs between ospC genes from the same Borrelia species (i.e. B. afzelii and B. garinii) as well as between different Borrelia species (i.e. B. afzelii and B. garinii, B. burgdorferi and genogroup DN127). PMID- 8709846 TI - pAM beta 1 resolvase has an atypical recombination site and requires a histone like protein HU. AB - The broad-host-range plasmid pAM beta 1 from Gram-positive bacteria encodes a resolvase, designated Res beta, which shares homology with the proteins of the resolvase-invertase family. Here we report the purification and in vitro characterization of Res beta. This resolvase is particular in two aspects: it has an atypical binding site and requires a cofactor to promote resolution in vitro. Res beta binds to two regions within its resolution site res. One contains two inverted repeats (R1 and R2), the other contains only one repeat (R3). The cofactor required for resolution in vitro is present in crude extracts of both Bacillus subtilis and Escherichia coli and can be substituted by the E. coli histone-like protein HU. The possible mode of action of HU in the resolution process is discussed. PMID- 8709848 TI - Bacillus subtilis MrgA is a Dps(PexB) homologue: evidence for metalloregulation of an oxidative-stress gene. AB - Upon the cessation of exponential growth, Bacillus subtilis enters a transition phase leading to either sporulation or a non-sporulating stationary phase. During this transition period, cells secrete degradative enzymes, become competent for DNA transformation, are motile and acquire resistance to oxidative killing. We now report that mrgA, originally identified as a gene repressed by metal ions, encodes a member of the Dps/PexB family of general stress proteins. Like Escherichia coli Dps(PexB), MrgA forms highly stable, multimeric protein-DNA complexes which accumulate in stationary-phase cells and protect against oxidative killing. MrgA is part of an inducible oxidative stress response in B. subtilis: mrgA is induced by hydrogen peroxide, and a strain lacking MrgA displays increased sensitivity to oxidative killing. In addition, a hydrogen peroxide-resistant mutant, which constitutively overproduces catalase and alkyl hydroperoxide reductase, also overproduces MrgA. These results indicate a complex interplay between metal ions and the expression of the B. subtilis oxidative stress response. PMID- 8709847 TI - Identification of mycoplasma membrane proteins by systematic Tn phoA mutagenesis of a recombinant library. AB - Wall-less prokaryotes in the genus Mycoplasma include over 90 species of infectious agents whose pathogenicity for humans and other animals is currently being assessed. Molecular characterization of surface proteins is critical in this regard but is hampered by the lack of genetic systems in these organisms. We used TnphoA transposition to systematically mutagenize, in Escherichia coli, a genomic plasmid library constructed from Mycoplasma fermentans, a potential human pathogen. The strategy circumvented problems of expressing mycoplasma genes containing UGA (Trp) codons and relied on the construction of the vector pG7ZCW, designed to reduce TnphoA transposition into vector sequences. Functional phoA gene fusions directly identified genes encoding 19 putative membrane-associated proteins of M. fermentans. Sequences of fusion constructs defined three types of export sequence: (1) non-cleavable, membrane-spanning sequences, (2) signal peptides with signal peptidase (SPase) I-like cleavage sites, and (3) signal peptides with SPase II-like lipoprotein-cleavage sites which, like most other mycoplasmal lipoprotein signals analysed to date, differed from those in several Gram-negative and Gram-positive eubacteria in their lack of a Leu residue at the 3 position. Antibodies to synthetic peptides that were deduced from two fusions to predicted lipoproteins, identified corresponding amphiphilic membrane proteins of 57 kDa and 78 kDa expressed in the mycoplasma. The P57 sequence contained a proline-rich N-terminal region analogous to an adhesin of Mycoplasma gallisepticum. The P78 protein was identical to a serologically defined phase variant surface lipoprotein. TnphoA mutagenesis provides an efficient means of systematically characterizing functionally diverse lipoproteins and other exported proteins in mycoplasmas. PMID- 8709849 TI - sbmC, a stationary-phase induced SOS Escherichia coli gene, whose product protects cells from the DNA replication inhibitor microcin B17. AB - Microcin B17 (MccB17) is a ribosomally synthesized peptide antibiotic of 43 amino acids that induces double-strand breaking of DNA in a DNA gyrase-dependent reaction. As a consequence, the SOS regulon is induced and massive DNA degradation occurs. In this work we have characterized an Escherichia coli gene, sbmC, that in high copy number determines high cell resistance to MccB17. sbmC encodes a cytoplasmic polypeptide of 157 amino acids (M(r), 18,095) that has been visualized in SDS-polyacrylamide gels. The gene is located at min 44 of the E. coli genetic map, close to the sbcB gene. sbmC expression is induced by DNA damaging agents and, also, by the entry of cells into the stationary growth phase. A G-->T transversion at the fifth nucleotide of the quasicanonical LexA box preceding the gene makes recA cells 16-fold more resistant to exogenous MccB17. The gene product, SbmC, also blocks MccB17 export from producing cells. Altogether, our results suggest that SbmC recognizes and sequesters MccB17 in a reversible way. PMID- 8709850 TI - Characterization of an Escherichia coli rotA mutant, affected in periplasmic peptidyl-prolyl cis/trans isomerase. AB - The rotA gene of Escherichia coli encodes a peptidyl-prolyl cis/trans isomerase (PPIase), which is supposed to catalyse protein folding in the periplasm. To investigate the importance of the enzyme, the rotA gene was cloned and a chromosomal deletion mutant was created. The rotA mutant was normally viable. No residual PPIase activity could be detected in the periplasmic fraction of the mutant. Comparison of the patterns of periplasmic and outer membrane proteins by SDS-PAGE revealed no differences in protein composition between the rotA mutant and its parental strain. Similarly, the kinetics of periplasmic protein folding and outer membrane protein assembly appeared unaffected by the rotA mutation. Our results show that the periplasmic PPIase of E. coli is not essential and that the protein does not play an important role in protein folding. PMID- 8709851 TI - Deletion analysis of gene minE which encodes the topological specificity factor of cell division in Escherichia coli. AB - Division inhibition caused by the minCD gene products of Escherichia coli is suppressed specifically at mid-cell by MinE protein expressed at physiological levels. Excess MinE allows division to take place also at the poles, leading to a minicell-forming (Min-) phenotype. In order to investigate the basis of this topological specificity, we have analysed the ability of truncated derivatives of MinE to suppress either minCD-dependent division inhibition in a chromosomal delta(minB) background, or the division inhibition exerted by MinCD at the cell poles in a minB+ strain. Our results indicate that these two effects are not mediated by identical interactions of MinE protein. In addition, gel filtration and the yeast two-hybrid system indicated that MinE interacts with itself by means of its central segment. Taken together, our results favour a model in which wild-type MinE dimer molecules direct the division inhibitor molecules to the cell poles, thus preventing polar divisions and allowing non-polar sites to divide. This model explains how excess MinE, or an excess of certain MinE derivatives which prevent the accumulation of the division inhibitor at the poles, can confer a Min- phenotype in a minB+ strain. PMID- 8709852 TI - Fnr activates transcription from the P6 promoter of the pfl operon in vitro. AB - Expression of the Escherichia coli focA-pfl operon is induced by anaerobiosis and transcription is controlled by seven promoters. Anaerobic induction is mediated by the ArcA and Fnr transcription factors. Fnr was purified and its specific interaction with the pfl promoter-regulatory region was characterized. A single binding site could be identified by DNase I footprinting, which was centred at 40.5 bp relative to the start site of promoter 6 (P6) transcription. Activation of P6 transcription in vitro was completely dependent on the Fnr protein. Fnr dependent transcription could be prevented by mutating the Fnr-binding site, indicating that Fnr must bind to its recognition sequence to be able to activate transcription. An Fnr-independent promoter, which overlaps the P6 promoter, was also identified and characterized in vivo and in vitro. Transcription from this promoter (termed P6A) initiated 10 bp upstream of the P6 start site and assures that low-level synthesis of the FocA protein occurs under aerobic growth conditions. PMID- 8709853 TI - Mutational analysis of the Yersinia enterocolitica virC operon: characterization of yscE, F, G, I, J, K required for Yop secretion and yscH encoding YopR. AB - Pathogenic yersiniae secrete the Yop anti-host proteins using a type-III secretion pathway. The components of the secretion machinery are encoded by three loci on the pYV plasmid: virA, virB, and virC. In this paper we describe the characterization of eight non-polar mutants of the virC locus, constructed by allelic exchange. The yscE, FG, I, J and K mutants were defective in Yop secretion and independent of Ca2+ (CI) for their growth at 37 degrees C. Substitution of the 12 N-terminal amino-acid residues of YscF impaired secretion of YopB and YopD only and led also to a CI phenotype. The culture supernatant of the yscH mutant contained all the Yops except the 18 kDa YopR. Complementation experiments and an immunoblot analysis confirmed that YopR is encoded by the yscH gene. The LD50 for the mouse of the yscH mutant was 10-fold higher than that of the parental strain indicating that YopR is involved in pathogenesis. The phenotype of the yscM mutant was similar to that of the wild-type strain. However, overproduction of YscM from a multicopy plasmid in wild-type Yersinia enterocolitica prevented Yop secretion and synthesis. A hybrid YopH-LacZ' protein, encoded by a gene transcribed from the lac promoter, was secreted by a strain overexpressing YscM, showing that the secretion machinery was still functional. These results indicate that YscM plays a role in the feedback inhibition of Yop synthesis when secretion is compromised by acting as a negative regulator of Yop synthesis. PMID- 8709855 TI - Why sequence genomes? The Escherichia coli imbroglio. PMID- 8709854 TI - Distinct and differently regulated Mo-dependent nitrogen-fixing systems evolved for heterocysts and vegetative cells of Anabaena variabilis ATCC 29413: characterization of the fdxH1/2 gene regions as part of the nif1/2 gene clusters. AB - Two different fdxH genes (fdxH1, fdxH2) have been isolated from the nitrogen fixing, heterocyst-forming cyanobacterium Anabaena variabilis ATCC 29413. They are part of two different nif gene clusters, nif1 and nif2. fdxH1 encodes the [2Fe-2S] ferredoxin that is known as the direct electron donor to nitrogenase in heterocysts, and is very similar to FdxH from Anabaena sp. PCC 7120. FdxH2 has more residues in common and shares its oxygen sensitivity with the single FdxH from the non-heterocystous, filamentous cyanobacterium Plectonema boryanum PCC 73110. The latter expresses nitrogenase early (< or = 3-4h) after nitrogen depletion in vegetative cells and exclusively under anaerobic conditions. fdxH2 and the nif2 genes of Anabaena 29413 are also transcribed < or = 4 h after onset of nitrogen-stepdown, exclusively under anaerobic growth conditions and long before functional heterocysts appear. At this time, no fdxH1 and nif1 gene transcription was observed. It occurred later and was associated with nitrogen fixation under aerobic conditions, i.e. within heterocysts. fdxH2 and nifHDK2 were not transcribed during aerobic, nitrogen-fixing growth. In addition, neither was an fdxH2-type gene found nor an anaerobically and early inducible Nif2 system detectable in Anabaena 7120. These data reveal that in filamentous cyanobacteria two different Nif systems have evolved based on molybdenum nitrogenases. It is concluded that a Nif2-type system operates in vegetative cells of non heterocystous and some, but not all, heterocyst-forming filamentous cyanobacteria. It is environmentally regulated by the levels of both oxygen and combined nitrogen in the habitat. To simultaneously allow for oxygen-evolving photosynthesis and oxygen-sensitive nitrogen fixation, the Nif1-type system probably branched from an ancestral Nif2-type system and has evolved for an exclusive operation within heterocysts. Accordingly, its expression has become an obligate late event in the developmental programme of heterocyst differentiation, irrespective of aerobic or anaerobic growth conditions. PMID- 8709856 TI - The Pasteurella multocida txaR gene is expressed in Escherichia coli but does not repress transcription of the P. multocida toxin gene toxA. PMID- 8709857 TI - Common structural features of IgA1 protease-like outer membrane protein autotransporters. PMID- 8709858 TI - Qid3 protein links plant bimodular proteins with fungal hydrophobins. PMID- 8709859 TI - Isolation and characterization of teichoic acid-lake substance as an adhesin of Staphylococcus aureus to HeLa cells. AB - A cell wall component that bound to HeLa cells (HeLa cell-binding CWC) was isolated from a clinical isolate of Staphylococcus aureus. The HeLa cell-binding CWC was resistant to heat (100 C, 1 hr) and proteases, did not stain with Coomassie Brilliant Blue R-250 on SDS-PAGE but stained as a broad band with antiserum against the strain on Western blots. These data suggest that the HeLa cell-binding CWC is not a protein, and may be teichoic acid. Purified teichoic acid bound to HeLa cells, whereas fractions without teichoic acid did not. In Western blots, HeLa cell-binding CWC appeared as a broad band of less than 35 kDa, similar to that of purified teichoic acid. These data suggest that the HeLa cell-binding CWC obtained in this study is teichoic acid. Teichoic acid inhibited S. aureus adherence to HeLa cells and bound to the cells time and dose dependently, in a saturable and reversible manner, and therefore appears to be an adhesin of S. aureus to HeLa cells. PMID- 8709860 TI - Analysis of the phospholipase C gene of Clostridium perfringens KZ1340 isolated from Antarctic soil. AB - Clostridium perfringens KZ1340 isolated from Antarctic soil was first classified as Clostridium plagarum and later as a lecithinase-negative variant of C. perfringens. Although the strain produced no detectable lecithinase (phospholipase C, PLC) activity in the culture supernatant, it was shown by Southern blot hybridization to possess a PLC-encoding gene (plc). To determine the cause of the PLC deficiency, we cloned and sequenced the plc gene from KZ1340. The deduced amino acid sequence consists of 398 amino acid residues, coinciding with those of the plc genes previously determined. Tyrosine was substituted for histidine at amino acid position 148, which is thought to bind a zinc ion essential for PLC activity. Northern blot analysis revealed that KZ1340 expressed the plc gene at an extremely low level. Furthermore, the plc gene cloned from C. perfringens strain 13 into a plasmid was expressed weakly in KZ1340, compared to that in strain 13. This indicates that the former strain represses plc gene expression in trans. When a phylogenetic tree of plc genes was constructed, the KZ1340 plc gene formed a monophyletic branch along with those of various other C. perfringens strains, supporting the classification of the strain as a variant of C. perfringens. PMID- 8709861 TI - Experimental Chlamydia psittaci infection of Japanese quail. AB - Japanese quail were used for the infection model of avian chlamydiosis. One-day old Japanese quail were highly susceptible to lethal infection by a Chlamydia psittaci strain of budgerigar origin upon inoculation via the air sac route with 10(4.1) FFU of the organism, showing an acute and lethal course with chlamydial propagation. In contrast, 7-day-old quail developed resistance to the infection as shown by the lack of lethal effect with the same dose. The resistance of 7-day old birds was abolished by immunosuppressive treatment with cyclophosphamide. Upon inoculation with a sublethal dose of 10(2.1) FFU, latent infection was established in 1-day-old birds with a minimum number of the organism. The latent infection in the birds was converted to the lethal form by treatment with cyclophosphamide along with chlamydial propagation and suppression of antibody production. PMID- 8709862 TI - Isolation and characterization of mumps virus strains in a mumps outbreak with a high incidence of aseptic meningitis. AB - In 1993, mumps with a high incidence of aseptic meningitis became prevalent in Akita prefecture, Japan. Three mumps virus isolates obtained from the nonvaccine associated cases lacked the BamHI restriction cleavage site of the P gene, like the Urabe strain (Yamada, A. et al, Vaccine 8: 553-557). However, four additional nucleotide substitutions were found in the determined region of 157 bp. Fourteen of 19 cases from which mumps virus showing the Urabe-like RFLP profile was detected were complicated with symptomatic meningitis, whereas there were only four cases of meningitis among 23 individuals infected with the wild type showing no Urabe-like RFLP profile (non-"Urabe-like" wild-type). The incidence of meningitis was over 70% among patients infected with the "Urabe-like" wild-type virus. The "Urabe-like" wild-type. disappeared after February 1994 in the epidemic area and was replaced by the non-"Urabe-like" wild-type. Patients infected with the "Urabe-like" wild-type lived in a closed colony, in which there were two instances of transmission between siblings. Thus this outbreak was transient and narrowly localized. PMID- 8709863 TI - Relationship of HIV-1 envelope V2 and V3 sequences of the primary isolates to the viral phenotype. AB - We examined the relationship between the amino acid sequences of the V2 and V3 regions of the envelope protein and the biological properties of ten human immunodeficiency virus type 1 (HIV-1) primary isolates. The infectivity, cytopathic effect (CPE), and syncytium forming activity of these primary isolates were tested against three T cell lines (CEM, MT2, and MOLT4/CL.8 cells), CD8 depleted peripheral blood mononuclear cells (PBMC), and primary monocyte-derived macrophages (MDM) from seronegative donors. In addition to the viral groups which had the syncytium inducing/T-cell line tropic (SI/TT) phenotype or non-syncytium inducing/non-T cell line tropic (NSI/NT) phenotype (including the NSI/macrophage tropic (NSI/MT) phenotype), there was a group of viruses that infected one or two T cell lines and PBMC but could not mediate syncytium formation. We therefore classified this group of viruses as a non-syncytium inducing/partial T-cell line tropic (NSI/pTT) virus. To investigate the relationship between these viral phenotypes and the sequence variability of the V2 and V3 regions of the envelope, we cloned the viral gene segment and sequenced the individual isolates. The sequence data suggested that the SI/TT type changes in the V3 sequence alone mediate a partial T cell line tropism and mild cytopathic effect and that an isolate became more virulent (SI/TT phenotype) if there were additional changes in the V2 or other regions. On the other hand, sequence changes in the V2 region alone could not mediate phenotypic changes but some additional changes in the other variable regions (for example, V3) might be required for the phenotypic changes in combination with changes in V2. These findings also suggested that amino acid changes in both the V2 and V3 region are required for the development of virulent variants of HIV-1 that outgrow during advanced stages of the disease. PMID- 8709864 TI - Immunohistochemical studies on the transneuronal spread of virulent herpes simplex virus type 2 and its US3 protein kinase-deficient mutant after ocular inoculation. AB - The transneuronal spread of a virulent wild-type herpes simplex virus type 2 (HSV 2) and its US3 protein kinase-deficient (US3 PK-) mutant was immunohistochemically studied in mice after inoculations into the cornea, anterior chamber, tongue, and masseter muscle. After corneal inoculation, the wild-type virus was demonstrated in various brain stem areas including the trigeminal tract and nucleus, the reticular formation, and cerebellar nucleus group. Viral antigen-positive neurons were strictly confined to the ipsilateral spinal trigeminal nucleus in mice corneally infected with the US3 PK- mutant. No viral antigens were detected in the central nervous system (CNS) after inoculation with the mutant into the tongue and masseter muscle. However, when mice were immunosuppressed by treatment with cyclophosphamide, both the corneally infected mutant and wild-type virus could invade the CNS. The results suggest that the US3 PK- mutant principally retains the capacity to spread in the CNS. PMID- 8709865 TI - Evidence that two distinct populations of rabbit anti-idiotypic antibodies are induced by three monoclonal antibodies specific for bluetongue virus core protein VP7. AB - Three groups of anti-idiotypic antibodies (anti-Id or Ab2), designated RAb2-A, RAb2-B1, and RAb2-B2, were isolated from rabbit antiserum raised against three monoclonal antibodies (mAbs) (M1875, M1877, and M1886) specific for the bluetongue virus core protein, VP7. RAb2-A was specific for the idiotype of M1875. RAb2-B1 and RAb2-B2, isolated through the M1877 and M1886 affinity columns, respectively, were directed against the common idiotype that is shared by M1877 and M1886 and therefore classified in the same population (RAb2-B). Further characterization revealed that the two anti-Id populations, RAb2-A and RAb2-B, were significantly different. RAb2-A was an Ab2 beta type of anti-Id since (i) its reaction with M1875 was inhibited by the antigen; (ii) it inhibited the M1875-VP7 interaction; and (iii) it elicited anti-VP7 antibody response in Balb/c mice. In contrast, RAb2-B may represent an Ab2 alpha type of anti-Id since its reactions with M1877 or M1886 were not inhibited by the antigen, even though it inhibited mAbs from binding to the antigen. These results indicated that RAb2 A and RAb2-B represent two distinct populations of anti-Ids to anti-VP7 mAbs with similar epitope specificity. PMID- 8709866 TI - Analysis of Vibrio cholerae O139 Bengal isolated from different geographical areas using macrorestriction DNA analysis. AB - Vibrio cholerae O139 isolated from different countries, as well as from different locations within a country, were examined using macrorestriction DNA analysis to determine the clonality of the O139 strains. NotI digests of genomic DNA of representative strains from Nepal, India, Bangladesh, China, Thailand, and Malaysia revealed very similar but not identical patterns. Examinations of the banding patterns generated by pulsed-field gel electrophoresis of strains isolated within countries revealed complete homogeneity. These results further reiterate the spread of an identical clone of V. cholerae O139 although it appears that genetic polymorphism among the O139 strains is becoming apparent. PMID- 8709867 TI - Distribution of Borrelia species associated with Lyme disease in the subalpine forests of Nagano prefecture, Japan. AB - We surveyed the natural distribution of Borrelia species associated with Lyme disease in the subalpine forests of Nagano prefecture, Japan, during 1993-94. Tick-derived isolates (n = 112) from Ixodes persulcatus and rodent-derived isolates (n = 55) from Apodemus argenteus, Apodemus speciosus, Eothenomys andersoni, Eothenomys smithii, and Microtus montebelli were classified by rRNA gene restriction fragment length polymorphism analysis (RFLP ribotyping). Ribotype group IV (an intraspecific variant of Borrelia garinii) was predominant among the tick isolates. It was also isolated repeatedly from the rodents. Ribotype group III (Borrelia afzelii) was detected in low frequencies among the tick and rodent isolates. The data suggest that humans are likely to be exposed to the group IV when they are bitten by I. persulcatus ticks. PMID- 8709868 TI - Seroepidemiological survey of lymphocytic choriomeningitis virus in wild house mice in China with particular reference to their subspecies. AB - Serum samples from 337 wild house mice (Mus musculus) from 35 sites in China, collected in 1992 and 1993, were examined for antibodies against lymphocytic choriomeningitis virus (LCMV). Ten samples from eight sites were found to contain such antibodies. Six of the eight positive sites were located in the territory of M. m. gansuensis. One of the other two sites was located in the territory of M. m. castaneus in southern China and the other site was in a habitat of M. m. castaneus which had invaded into the western end of the territory of M. m. homourus. It seems likely that LCMV is distributed in the territories of M. m. gansuensis and M. m. castaneus in China. This is the first report of detection of these antibodies in wild house mice in China and specifically in the territories of M. m. gansuensis and M. m. castaneus. PMID- 8709870 TI - Urinary incontinence: the Cinderella subject. PMID- 8709869 TI - Detection and sequencing of Norwalk-like viruses from stool samples in Japan using reverse transcription-polymerase chain reaction amplification. AB - Norwalk-like viruses were detected in Japan in 12% (26/209) of patients with nonbacteria and nonrotavirus gastroenteritis in an outpatient clinic of a hospital from 1991 to 1994 by reverse transcription-polymerase chain reaction. They were also present in 7% (26/378) of total samples including those from rotavirus-positive gastroenteritis patients. In addition, the viruses were recovered in samples from 15 of 17 patients which were collected during outbreaks of gastroenteritis in various places in Japan by the same method. The DNA sequence of the polymerase region from patients at the hospital (sporadic cases) showed that two subgenogroups, similar to UK1-6 in genogroup G1 and to UK1-1 in genogroup G2 (Ando et al, J Clin Microbiol, 1995, 33: 64-71) exist in Japan. The latter was more frequently found. PMID- 8709871 TI - Ensuring quality in all phases of the pathology cycle. PMID- 8709872 TI - Renal disease in Australian aboriginals. PMID- 8709873 TI - Error rates in Australian chemical pathology laboratories. AB - OBJECTIVE: To measure transcription and analytical errors made by Australian chemical pathology laboratories. DESIGN: Retrospective data collection covering the period 1 November 1993 to 1 April 1994. SETTING AND PARTICIPANTS: Fourteen pathology laboratories in five Australian States (seven in the public sector, and seven in the private sector). MAIN OUTCOME MEASURES: Error rates in transcribing information from request forms to computer record systems, and laboratory performance on chemical analysis. RESULTS: Pathology laboratories had a transcription-error rate of up to 39% and an error rate of up to 26% for analytical results. The worst-performing laboratory had errors (of patient identification or results of analysis) in 46% of requests. The three best performing laboratories achieved 85% error-free reporting, with one achieving 95%. CONCLUSIONS: Error rates in Australian pathology laboratories vary widely, but may be as high as 46% for all specimens in some laboratories. The types of errors reported were under the control of the laboratory, and would affect the accuracy of reported pathology test results, with potential adverse outcomes for patient care and inefficient use of health-care resources. There is a need to establish broader quality assurance programs and performance requirements to reduce these types of error. PMID- 8709874 TI - Barriers to effective primary health care for adolescents. AB - OBJECTIVES: To assess general practitioners' (GPs') perceptions of barriers in the health care system that hinder provision of effective adolescent health care; and to assess the training needs of GPs for a future education program in adolescent health issues. DESIGN: Retrospective questionnaire survey. METHODS: A 66-item questionnaire was mailed during May and June 1995 to a random stratified sample of 997 rural and urban Victorian GPs. A Practice Assessment Task enabling respondents to fulfil Royal Australian College of General Practitioners' quality assurance requirements was mailed on return of completed questionnaires if requested. RESULTS: The response rate was 72.4%. After adjusting for stratified sampling, 77% (95% confidence interval [CI], 73%-81%) of respondents favoured individual Medicare cards for adolescents from the age of 16. Thirty-eight per cent (95% CI, 34%-43%) were less willing to charge for longer consultations because they feared Health Insurance Commission (HIC) investigation. Most respondents indicated that their undergraduate training in adolescent mental health issues was inadequate and 64% (95% CI, 61%-68%) found it difficult to obtain advice about complex mental health problems. An interest in continuing medical education in adolescent health issues was expressed by 82% (95% CI, 79% 86%) of respondents. CONCLUSIONS: Individual Medicare cards should be automatically issued to adolescents from the age of 16 to improve their ability to access health care. Revision of the Medicare rebate system and clarification of HIC's investigative functions may improve GP's effectiveness in adolescent consultations. Undergraduate medical and GP training should include curriculum relevant to adolescent health care. PMID- 8709875 TI - School bus related deaths and injuries in New South Wales. AB - OBJECTIVE: To report the circumstances of paediatric bus related deaths and injuries in New South Wales (NSW) to identify preventable factors. DESIGN: Retrospective survey using two surveillance systems-the NSW Paediatric Death Review Database and the Childsafe Injury Surveillance System. PATIENTS: Children (0-14 years of age) who died or were injured as a result of a school bus related incident. RESULTS: Twenty-two deaths and 58 injuries were recorded. Three of the children who died were passengers (two deaths were due to errant behaviour), two were alighting from the bus (bus door entrapment) and 17 (77%) were pedestrians crossing the road before or as the bus departed. The major causes of death were head injury and blood loss. Seventeen of the injured children were pedestrians and most (82%) of these sustained serious injury requiring admission to hospital. CONCLUSIONS: The greatest risk to schoolchildren from bus related injuries was as pedestrians after alighting from a bus, especially when moving behind the bus. Preventable factors include slowing of traffic (40 kph) near stationary school buses, and parents waiting on the side of the road where the child alights from the bus. Continuing road safety education remains important for schoolchildren and parents as well as for drivers. PMID- 8709876 TI - Accidental infant death and stroller-prams. AB - A three-month-old boy and an eight-month-old boy died from accidental positional asphyxia and hanging, respectively, after being placed to sleep unsupervised in stroller-prams. Both infants had moved down towards the fronts of the stroller prams. The younger infant fell out when the footplate collapsed and he was found hanging from a metal bar on the side. The older infant had partly slipped through the front and was suspended with his head and arms within the stroller-pram and with his face pushed firmly into the mattress by a horizontal metal bar. Stroller prams are a potentially dangerous sleeping environment unless infants are closely supervised, gaps in the front of stroller-prams closed and upright footplates stabilised. PMID- 8709877 TI - Managing HIV. Part 7: Professional issues. 7.4 HIV and complementary medicine. AB - Many people with HIV use complementary or alternative therapies, often without the knowledge of their doctors. To encourage open communication with their patients, doctors need to be aware of what complementary therapies are available and willing to discuss their use. They are then in a better position to defend their patients against exploitation or unsafe treatment. PMID- 8709878 TI - Managing HIV. Part 7: Professional issues. 7.5 HIV and choosing to die. AB - People with HIV have been encouraged to take control of their health and the monitoring and treatment of their illnesses. Many claim the right to choose the time and manner of death. PMID- 8709879 TI - Managing HIV. Part 7: Professional issues. 7.6 Legal issues in HIV medicine. AB - The mortality, stigma and publicity surrounding HIV/AIDS have brought into focus many legal and ethical issues arising in the practice of medicine. Three important aspects of the practice of HIV medicine are the obligation to notify health authorities of HIV diagnoses, special obligations to inform and counsel patients who have or may have HIV and the duty of confidentiality. PMID- 8709880 TI - Managing HIV. Part 7: Professional issues. 7.7 HIV care: the impact on the doctor. AB - Managing patients with HIV raises complex clinical, ethical and personal challenges. "Burnout" threatens the overcommitted doctor who is not prepared to take preventive measures. PMID- 8709881 TI - Managing HIV. Part 7: Professional issues. 7.8 Continuing HIV education for health care workers. AB - A five-year national education program for HIV/AIDS has brought into focus many issues in continuing education and has evolved models worthy of consideration by colleagues in other fields. PMID- 8709882 TI - Acne: what's new? PMID- 8709883 TI - Is a general practitioner legally bound to render assistance to a stranger? PMID- 8709884 TI - Oral retinoids and pregnancy. AB - The oral retinoids isotretinoin and etretinate are uniquely effective in the treatment of severe cystic acne and keratinisation disorders. Because of their known teratogenicity, there are strict prescription guidelines, but exposure during pregnancy still occurs. A dedicated effort by women and their clinicians is required, involving patient selection, education and informed consent, detailed contraceptive counselling, and careful monitoring and management, including pregnancy testing before commencement of therapy. PMID- 8709885 TI - Kidney transplantation from living non-related donors. PMID- 8709886 TI - Teaching hospital medical staff to handwash. PMID- 8709887 TI - Suspected snakebite in children: a study of 156 patients over 10 years. PMID- 8709888 TI - Attention-deficit disorder with hyperactivity. PMID- 8709889 TI - Vitamin B12 myeloneuropathy precipitated by nitrous oxide anaesthesia. PMID- 8709890 TI - Screening for osteoporosis: what is the role of heel ultrasound? PMID- 8709891 TI - Screening for osteoporosis: what is the role of heel ultrasound? PMID- 8709892 TI - The safety of topical beta-blockers in glaucoma treatment. PMID- 8709893 TI - Conceptualising the outcomes of war exposure. PMID- 8709894 TI - Limited adverse event screening: using medical record review to reduce hospital adverse patient events. PMID- 8709895 TI - Teleconsulting. PMID- 8709896 TI - [Smoke-free zones]. PMID- 8709897 TI - [Tobacco smoking cessation. Methods and aids]. PMID- 8709898 TI - [Endangering health by smoking. Results of an English long-term study]. PMID- 8709899 TI - [From CARE and CHAOS to HERO and PRIME. Report of the 45th Annual Meeting of the American College of Cardiology, Orlando, 24-27 March 1996]. PMID- 8709900 TI - [Improving bladder function by pumpkin seeds?]. PMID- 8709901 TI - [Positions and perspectives. Opening address by the chairman of the 102nd Congress of the German Society of Internal Medicine, Wiesbaden, 14 April 1996]. PMID- 8709902 TI - [European codex against cancer]. PMID- 8709903 TI - [Follow-up and immunologic findings in drug-induced myasthenia]. AB - PATIENT AND METHOD: We report 5 patients, who developed myasthenia, four of them after treatment on D-penicillamine, one after treatment on chloroquine. 3 patients suffered from rheumatoid arthritis, one from a psoriatic arthritis and one from cirrhosis of the liver. Three patients developed an ocular myasthenia, one patient an oculopharyngeal and one patient has had generalized myasthenic syndrome. RESULTS: Four patients showed an improvement of clinical status within days up to 18 months following discontinuation of the therapy, whereas one patient deteriorated. The quickest improvement was observed in the patient with chloroquine induced myasthenia. Four patients had raised acetylcholine-receptor antibody titers. The patient with chloroquine induced myasthenia had had a normal acetylcholin-receptor antibody titer. The human leucocyte antigen type was compared with the results of literature. CONCLUSION: To what extent human leucocyte antigen type HLA DR 4 has a correlation with the development of myasthenia following treatment on chloroquine can not yet be answered with respect to the very small number of cases at the moment. PMID- 8709904 TI - [Attitude of established physicians to treating HIV-infected patients and to "acquired immunodeficiency syndrome". An empirical study]. AB - BACKGROUND: There is no empirical data available on attitudes concerning AIDS and habits towards HIV infected patients of physicians in general or private practice. In this study results of a self-evaluation are presented. METHODS: 178 physicians working with out-patients in different medical fields were randomly selected for a cross sectional study and interviewed using a standardised questionnaire. RESULTS: 89% think that they are sufficiently informed about AIDS (in the USA 20%). They regarded the risk of infection to be lower than the Anglo American physicians. They believed there is a lack of interchange of information between colleagues regarding the degree of infectiousness of referred patients. A third of the physicians fear that other patients will go elsewhere if they find out that their physician is treating AIDS patients. 54% would hold special clinic sessions for HIV-patients outside the normal schedule for practice times. 89% believed that HIV patients were partly to blame for their illness. CONCLUSIONS: Although the physicians recognise the problem of HIV-infection, they partly deny the real necessities and facts. A reason for this could be the emotions underlying the general attitude to everything pertaining to HIV-disease. Attitudes to HIV-disease and the dealing with it in daily practice must be considered on the basis of individual emotional motives. PMID- 8709905 TI - [Clinical studies on combination of ifosfamide, carboplatin and etoposide (ICE) with whole body hyperthermia]. PMID- 8709906 TI - [Microsporidia--biology and clinical relevance]. PMID- 8709907 TI - [Therapeutic strategies in diabetic neuropathy. Pathogenetically founded therapeutic approaches in peripheral, sensory, symmetrical neuropathy]. PMID- 8709908 TI - [Limitations of current treatment of asthma: possibilities for new treatment forms]. PMID- 8709909 TI - [APC resistance: pathogenesis, diagnosis, prevalence and importance of a new thrombosis risk factor]. PMID- 8709910 TI - [Fever and pancytopenia in extrathoracic sarcoidosis]. PMID- 8709911 TI - [Public health economic evaluation of screening for APC resistance (Leiden mutation) in new oral contraceptive users]. AB - BACKGROUND: Thromboembolic events in users of oral contraceptives rank among the most important complications with potential economic consequences. It is well known that a large part of thromboembolic complications correlates with hereditary thrombophilias. The relative high prevalence of the newly described resistance of activated protein C and an easy test for it rise up the question if general screening of new users of oral contraceptives is sensible of health economic view. METHOD: We conducted a cost-effectiveness analysis using decision analytic techniques, analysing the costs and outcomes in a hypothetical cohort of 10,000 women. RESULT AND CONCLUSION: From a third party payer perspective we estimate that screening for APC resistance by new users of oral contraceptives is more cost-effective than many other primary preventive methods. The cost per life year gained of testing for APC resistance were in the order of DM 1544,- including direct medical cost only. From perspective of sickness insurance fund as cost unit we estimate that screening for APC resistance by new users of oral contraceptives is more cost-effective than many other primary preventive methods. PMID- 8709912 TI - [Early detection of colorectal carcinoma]. PMID- 8709913 TI - [Early detection of colorectal carcinoma]. PMID- 8709914 TI - [Differential diagnosis of Hantavirus infection--adult Still syndrome]. PMID- 8709915 TI - [Infectious origin of Whipple disease]. PMID- 8709916 TI - [Cryptorchidism: current views]. AB - Cryptorchidism is a pathological condition which affects up to 6% of newborns. Main etiopathogenetic hypotheses are the hormonal and the dysgenetic one. Ultrasonography is useful in locating testis in the inguinal canal, while in the management of intraabdominal testis, laparoscopy is considered the best diagnostic technique and, in many cases, can be coupled with surgical management. Medical treatment with LH-RH or with hCG or, better, combined (LH-RH+hCG) is recommended before the second year. Impairment of fertility is a complication mainly in subjects with a history of bilateral cryptorchidism. Undescended testis has a risk of malignant degeneration ranging from 3% to 18% and for this reason some authors suggest a gonadal biopsy after puberty. PMID- 8709917 TI - Time-relationship between bone growth and increment of bone mineral content in growing rats. AB - The aim of the study was to describe the time-relationship between the growth rate of the rat tibia and the increase rate of its mineral content. Appositional and endochondral bone growth rates were derived from sequential X-rays measurements of tibial widening and lengthening, respectively; the increase rate of bone mineral content was derived from sequential photon absorptiometry measurements of the proximal tibio-fibular site. The time-relationship of appositional growth rate and endochondral growth rate versus bone mineral content increase rate was mathematically described. The results allow a better understanding of the time-course of two distinct features of bone growth: increase in size and increase in mineral content. PMID- 8709918 TI - [Effects of prolonged administration of branched-chain amino acids on body composition and physical fitness]. AB - No conclusive data are reported about the effects of long-term branched-chain amino-acids (BCAA) administration on metabolic and respiratory parameters during the sustained hand-grip test (SHGT). Bedside, few data are reported about the relationship between BCAA administration and body composition. Thus, BCAA (14 g/die; 50% L-Leucine, 25% L-Isoleucine, 25% L-Valine) were administered orally to 10 untrained male healthy subjects for 30 days. Before (T0) and after (T1) BCAA administration, oxygen uptake (VO2) and pulmonary ventilation (VE) were evaluated at rest ("baseline", B) and during SHGT. Body height and body weight, body mass index (BMI), fat mass (FM), fat-free mass (FFM), arm muscle area (AMA) and maximal voluntary contraction of forearm muscler (GS, grip strength) were also evaluated at T0 and T1. At T1 FFM increased significantly (T0: 62.3 +/- 7.6 kg; T1: 63.1 +/- 7.5 kg; mean +/- SD; p < 0.05), as well as GS (T0: 46.8 +/- 7.9; T1: 50.6 +/- 7.8; p < 0.01). At T0, VO2 increased significantly during the test (B: 271 +/- 36 ml/min; SHGT: 332 +/- 71 ml/min; p < 0.01), whereas at T1 no significant increase was observed (B: 271 +/- 50 ml/min; SHGT: 286 +/- 60 ml/min). We conclude that the physical fitness of BCAA treated subjects was improved, because of the decrease of O2 uptake during SHGT. Moreover, in our subjects, BCAA seemed to promote FFM-protein synthesis mostly. PMID- 8709919 TI - [The use of color Doppler sonography in the diagnosis of thyroid pathology]. AB - Color Flow Doppler Sonography (CFD) is a technique that gives us useful information related to blood flow and consequently about some organs or tissues physiology and morphology. The purpose of this paper is to discuss some new developments on CFD's role in the diagnosis of thyroid disease, an application that has recently been put forward. Some studies using CFD have described, as in the case of other organs and tissues, the main qualities of gland's parenchyma vascularity both in physiological conditions and in the instance of diffuse and/or focal thyroid diseases. A large part of these studies have dealt with the aspects of hypervascularity in malignant thyroid neoplasms, focusing on the meaning of the intranodular hypervascularity in the differential diagnosis between benign and malignant thyroid lesions. CFD, although reported as having high sensibility and specificity, is not yet in a position to improve much on the current limits of fine-needle aspiration biopsy (FNAB), in particular in the case of "follicular proliferation" where the distinction between benign adenoma and thyroid carcinoma is still a frequent problem. Further studies are required to evaluate if CFD can reduce the percentage of patients to be operated upon because of unclear cytological pattern. PMID- 8709920 TI - A brain metastasis, with no uptake of 99mTc-ECD, secondary to Hurtle cell carcinoma of the thyroid. AB - Visualization of cerebral perfusion with Single Photon Emission Computed Tomography (SPET) needs lipophil radiotracers which are able to cross the intact blood-brain-barrier (BBB); moreover local uptake must reflect regional cerebral blood flow. In the last decade many radiopharmaceuticals have been suggested and employed for this purpose. Recently a new molecule has been synthesized: technetium-99m-labeled ethyl cysteinate dimer (99mTc-ECD). Commonly, SPET of the brain with perfusion tracers is performed to identify ischemic regions. We describe a patient with a bulky cerebral metastasis, secondary to Hurthle cell carcinoma of the thyroid, which concentrated radioiodine. Brain metastases secondary to differentiated carcinoma of the thyroid are very rare and when treated with 131-I may provoke serious neurological problems. Therefore we decided to perform a brain SPET with 99mTc-ECD to exclude regional perfusion defects and deafferentation phenomena. Brain CT showed only the large metastasis. SPET images, reconstructed according to standard tomographic planes of the head: transverse, coronal and sagittal revealed no uptake of 99mTc-ECD within the metastasis and no regional perfusion defects related to compression phenomena or deafferentation. 99mTc-ECD has never been used to image cerebral neoplasm of whatever origin. PMID- 8709921 TI - [Male pseudohermaphroditism due to 17-alpha-hydroxylase deficiency]. AB - A case of 17-alpha-hydroxylase deficiency in a genotypic male is described. The patient, who had grown up as a female, at the age of 18 years by chance was seen for enteritis. She presented primary amenorrhea and lack of secondary sexual characteristics combined with hypertension and mild hypokalemia due to excess of mineralcorticoids. The hormonal profile observed under basal conditions was evocative of a deficiency in 17-alpha-hydroxylase. High plasma aldosterone concentrations, in the face of suppressed PRA, were related to interference in RIA method: low plasma aldosterone values were observed when HPLC separation was applied. Eunucoid appearance and gigantism (195 cm) has rarely been observed as a fenotypic expression of this enzymatic deficit, but are justified by deficiency of both androgens and estrogens. PMID- 8709922 TI - [Insulin-dependent diabetes mellitus induced by interferon-alpha]. AB - The efficacy in viral neoplastic illness of IFN alpha therapy is responsible of an increasing use of this drug. Disease like hypothyroidism, thyroiditis, thyreotoxicosis, pernicious anemia, LES, vasculitis, thrombocytopenic purpura and, more recently, diabete mellitus, have been described during IFN alpha therapy. We describe another case of diabetes mellitus induced by IFN alpha. PMID- 8709923 TI - Prenatal coexposure to metallic mercury vapour and methylmercury produce interactive behavioural changes in adult rats. AB - Pregnant rats were 1) administered methyl mercury (MeHg) by gavage, 2 mg/kg/day during days 6-9 of gestation, 2) exposed by inhalation to metallic mercury (Hg degrees) vapour (1.8 mg/m3 air for 1.5 h per day) during gestation days 14-19, 3) exposed to both MeHg by gavage and Hg degrees vapour by inhalation (MeHg + Hg degrees), or 4) were given combined vehicle administration for each of the two treatments (control). The inhalation regimen corresponded to an approximate dose of 0.1 mg Hg degrees/kg/day. Clinical observations and developmental markers up to weaning showed no differences between any of the groups. Testing of behavioural function was performed between 4 and 5 months of age and included spontaneous motor activity, spatial learning in a circular bath, and instrumental maze learning for food reward. Offspring of dams exposed to Hg degrees showed hyperactivity in the motor activity test chambers over all three parameters: locomotion, rearing and total activity; this effect was potentiated in the animals of the MeHg + Hg degrees group. In the swim maze test, the MeHg + Hg degrees and Hg degrees groups evidenced longer latencies to reach a submerged platform, which they had learned to mount the day before, compared to either the control or MeHg groups. In the modified, enclosed radial arm maze, both the MeHg + Hg degrees and Hg degrees groups showed more ambulations and rearings in the activity test prior to the learning test. During the learning trial, the same groups (i.e., MeHg + Hg degrees and Hg degrees) showed longer latencies and made more errors in acquiring all eight pellets. Generally, the results indicate that prenatal exposure to Hg degrees causes alterations to both spontaneous and learned behaviours, suggesting some deficit in adaptive functions. Coexposure to MeHg, which by itself did not alter these functions at the dose given in this study, served to significantly aggravate the changes. PMID- 8709924 TI - Neonatal methamphetamine-induced long-term acoustic startle facilitation in rats as a function of prepulse stimulus intensity. AB - Neonatal exposure to methamphetamine (MA) has previously been shown to induce acoustic startle facilitation when the animals were tested as adults. The present experiment sought to replicate and extend this effect using a lower dose of MA and to determine if the effect varied as a function of prepulse stimulus intensity. Sprague-Dawley CD rat offspring were culled on the day after birth to eight (preferentially retaining females). On days 1-10, progeny were injected SC with either 20 mg/kg of d-MA twice per day (doses spaced at least 8 h apart) or distilled water. On postnatal day 50, offspring were administered 51 acoustic startle trials followed by 36 prepulse trials. Prepulse intensities were 0, 70, 75, 80, 85, or 90 dB. MA progeny showed augmented startle response amplitudes on both paradigms but the effect was most pronounced on the prepulse trials. Prepulse intensity interacted with MA treatment in that significant facilitation in the MA animals occurred on 0 and 70 dB prepulse trials but was only a trend (p < 0.10) on 75, 80, 85, and 90 dB trials. This implies that the effect of MA is most likely upon the basic startle reflex and not upon inhibitory pathways that modify startle reactivity. PMID- 8709925 TI - Neonatal exposure to anti-nerve growth factor antibodies affects exploratory behavior of developing mice in the hole board. AB - The aim of this study was to assess in developing mice whether the neutralization of endogenous NGF following ICV administration of anti-NGF antibodies (50 micrograms/2 microliters) on postnatal days 3, 6, 9, and 12 affected locomotor activity, exploratory behavior, and response to the cholinergic blocker scopolamine. In Experiments 1 and 2 activity and age-typical scopolamine effects were evaluated on PND 13 or 17 in an automated apparatus. No significant main effect of anti-NGF treatment was found at either age. On day 13 scopolamine (0.2, 1, or 2 mg/kg) decreased locomotion in both anti-NGF and control animals. In Experiment 3, locomotion and exploratory behavior were analyzed in an open field arena or in a hole board apparatus on PND 16. No significant effects of anti-NGF treatment on general motor activity and investigation of a novel object in the open field was found, though anti-NGF animals tended to be less active than controls. In the hole board anti-NGF pups showed a different pattern of head dipping behavior from controls, exploring mainly the holes located in the periphery of the apparatus. PMID- 8709926 TI - The effect of chronic cocaine exposure during pregnancy on maternal and infant outcomes in the rhesus monkey. AB - To explore the effects of gestational cocaine exposure in a nonhuman primate model, pregnant rhesus monkeys were treated from about 1 month of gestation until term with either 0 (N = 3), 0.3 (N = 3), 1.0 (N = 3), or escalating doses up to 8.5 (N = 3) mg/kg (IM), three times per day, 5 consecutive days per week. Despite these differences in cocaine exposure, the experimental groups did not differ significantly with respect to maternal outcome, as measured by body weight gain during pregnancy and length of pregnancy. A clear dose-response relationship was observed between the cumulative dose of cocaine administered during gestation and the levels of both cocaine and its major metabolite, benzoylecgonine, in samples of infant hair taken at birth. However, the experimental groups did not differ significantly with respect to infant outcome, as measured at birth by body weight, overall length, crown-to-rump length, rump-to-heel length, biparietal diameter, and crown circumference. Furthermore, the experimental groups did not differ significantly with respect to the integrity of a variety of infant reflexes tested at birth. It was concluded that, in a rhesus monkey model, chronic cocaine exposure during pregnancy had no significant effect on maternal and infant outcomes as assessed in this investigation. PMID- 8709927 TI - The effect of chronic cocaine exposure during pregnancy on the acquisition of operant behaviors by rhesus monkey offspring. AB - To explore possible long-term effects of gestational cocaine exposure in a nonhuman primate model, pregnant rhesus monkeys were treated from about 1 month of gestation until term with either 0 (N = 3), 0.3 (N = 3), 1.0 (N = 3), or escalating doses up to 8.5 (N = 3) mg/kg (IM), three times per day, 5 consecutive days per week. Despite these differences in cocaine exposure, the experimental groups did not differ significantly with respect to the postnatal growth of offspring over an 18-month period following birth. Beginning at 6 months of age, the behavior of offspring was monitored using an operant test battery that included five food-reinforced tasks designed to model aspects of learning, color and position discrimination, time estimation, short-term memory and attention, and motivation. Although the acquisition of each operant behavior by offspring progressed significantly during training between 6 and 18 months of age, this acquisition was not differentially affected by gestational cocaine exposure. It was concluded that, in a rhesus monkey model, chronic cocaine exposure during pregnancy had no significant effect on the offsprings' acquisition of operant behaviors. PMID- 8709928 TI - Toxocara canis infection in preschool age children: risk factors and the cognitive development of preschool children. AB - Risk factors for Toxocara canis (T. canis) infection were evaluated in a prospective study of disadvantaged preschool children. In addition, the hypothesis that T. canis exposure is associated with lower intelligence was tested. Seropositivity was tested at 2 years, 3 years, and at 4 years 10 months (4-10). Intelligence was measured at age 4-10 by the Full Scale IQ of the Wechsler Preschool and Primary Scales of Intelligence (WPPSI). Pica and ownership of a dog were unrelated to seropositivity. Seropositive children had lower scores on the Mental Development Index (MDI) of the Bayley Scales of Infant Development at age 1 year (prior to likely exposure). They also had less favorable scores on a measure of the quality of childrearing. These findings suggest that, for disadvantaged children, lower initial intelligence and less advantageous child rearing are risk factors for T. canis exposure. Seropositive children also had higher blood lead levels, probably as a result of the common pathway of hand to mouth transmittal. Seropositivity at 3 years, at age 4-10, or, cumulatively, at any of the age 2, 3, or 4-10 assessments was associated with the WPPSI IQ after adjustment for sociodemographic factors. Exposure at age 4-10-years was significantly associated with reduced IQ scores (p = 0.030). However, when the age 1 year MDI score was controlled, the estimate became nonsignificant. We, thus, can neither confirm nor deny a relationship of T. canis and intelligence, but the importance of considering prior developmental status is emphasized. PMID- 8709929 TI - In vitro exposure to aluminum does not alter long-term potentiation or glutamate release in rat hippocampal slices. AB - Aluminum has been reported to inhibit long-term potentiation (LTP) following in vivo administration and decrease glutamate release following in vitro exposure. Because glutamate release is critical for synaptic transmission and the development and maintenance of LTP in the hippocampus, we examined the effects of aluminum chloride (AlCl3) on depolarization-induced glutamate release and LTP in rat hippocampal slices. The effects of AlCl3 on [14C]glutamate release were examined by incubation of slices in depolarizing (56 mM)K+ buffer solution in the absence or presence of 2 mM CaCl2. After 15 min depolarization, AlCl3 (100-1000 microM) did not significantly affect Ca(2+)-dependent [14C]glutamate release from slices, whereas a known Ca2+ channel blocker (100 microM CdCl2) decreased Ca(2+) dependent [14C]glutamate release by approximately 50%. In contrast to a previous report, acute exposure to AlCl3 was without effect on depolarization-dependent glutamate release. LTP of the population spike (PS) in CA1 of hippocampus was induced by the delivery of stimulus trains to the stratum radiatum. LTP of the PS was observed in both control slices and slices bathed in solution containing 100 microM AlCl3. Neither the magnitude nor longevity (measured up to 1 h posttrain) of LTP distinguished control from aluminum-exposed slices. The lack of sensitivity in rat to the encephalopathic changes induced by aluminum, or methodological differences in exposure conditions may account for the lack of effect of aluminum on in vitro LTP in rat hippocampus. PMID- 8709930 TI - Effects of acute treatment with dopaminergic drugs on open field behavior of adult monkeys treated with lead during the first year postpartum. AB - A pharmacological challenge of nonhuman primate open field behavior, similar to that which previously assessed the cholinergic system (10), was used here to measure potential lead-induced alterations in the dopaminergic system. Monkeys that had been treated with lead during the first year postpartum were assessed at 7 years of age in the open field after acute intramuscular injection of apomorphine (0.0-0.3 mg/kg) or haloperidol (0.0-30 micrograms/kg). Duration of environmental exploration indicated a possible greater responsivity to 0.2 mg/kg apomorphine in the lead-treated monkeys; however, in all other behaviors, lead treated monkeys responded to both drugs similarly to controls. Regardless of lead treatment, apomorphine administration decreased duration of inactivity and increased environmental exploration; the latter possibly included an increase in stereotypical behavior that might have been recorded as environmental exploration. Haloperidol had no significant effects on open field behavior over the dose range tested. Open field behavioral alterations previously reported for these monkeys at 4-6 years of age were no longer strongly exhibited by the lead treated monkeys in any portion of the current study. Latency to enter the open field was marginally increased in the lead-treated group but levels of environmental exploration were comparable to controls. These results indicate an attenuation of lead-related effects with maturity and/or familiarity with the open field. PMID- 8709931 TI - Validation of automated behavioral test systems. AB - A positive control study was conducted as part of the ongoing validation program for developmental neurotoxicity testing in our laboratory using a standard battery of automated systems, consisting of rotorod, motor activity, acoustic startle, and two-way active avoidance. Female Sprague-Dawley rats were given 10 mg/kg diazepam (DZ) by SC injection or 20 mg/kg methimazole (MET) by gavage from gestation day 15 (DZ) or 17 (MET) through postpartum day 10; a group of control animals remained untreated. Offspring were assessed for growth, survival, developmental landmarks, and behavior. Although this study was considered useful for obtaining historical data, it offered few advantages in terms of validation of automated behavior test systems. Perinatal treatment with DZ resulted in no maternal toxicity and no adverse effects on growth or development of F1 offspring; a deficit in acoustic startle responding was the only behavioral effect observed. Treatment with MET resulted in maternal toxicity, reduced neonatal body weights, and developmental delays. Behavioral effects included impaired rotorod performance and acoustic startle responding (neonates), and enhanced motor activity and acoustic startle responding (young adults). However, effects on shuttle avoidance were not observed for either drug, and only one direction of behavioral effect occurred for the rotorod and motor activity systems. These results, as well as those from subsequent studies in our laboratory, suggest that it may be preferable to validate automated behavior systems using short-term studies in which young adult animals are treated directly with positive control agents. PMID- 8709932 TI - Stage-specific effects of prenatal d-methamphetamine exposure on behavioral and eye development in rats. AB - Pregnant Sprague-Dawley rats were treated with 0, 5, 10, 15, or 20 mg/kg d methamphetamine (MA), expressed as the free base, by SC injection (b.i.d., 8 h apart) on days 7-12 or 13-18 of gestation. Plasma concentration of MA and amphetamine were determined after the last dose. MA reduced gestation weight gain. The late exposure resulted in an increase in maternal and offspring mortality and reduced offspring growth. Offspring treated early in gestation with MA showed delayed development of early locomotion. In addition, memory impairment, evidenced by decreased target quadrant times and platform crossings on test trials and increased latency on reversal trials in the Morris spatial navigation maze, reduced spontaneous alternation, and lengthened passive avoidance retention latency was seen in the early treated high-dose groups. A reduction of serotonin was found in the nucleus accumbens following late exposure to MA at 20 mg/kg. Animals in both exposure groups had eye defects; however, the type of defect was dependent on the developmental stage at the time of dosing. Anophthalmia occurred only after early MA exposure, whereas folded retina was drug related only after late MA exposure. The behavioral effects did not show graded dose dependency; however, the effects were sensitive to exposure period. The early exposed animals had more alterations in behavior whereas the late exposed group showed higher mortality, reduced body weights, and neurochemical alterations. PMID- 8709933 TI - [Recent improvement of mammographic diagnosis]. AB - The improvement of mammographic diagnosis consists of 3 factors as follows; (1) improvement of mammographic apparatus, (2) technical advancement on photographing and reading of mammograms, and (3) correlation between mammographic findings and pathology. Improvement of apparatus was achieved by molybdenum anode and faster single screen combined with a high-speed single-emulsion film combination system. These system provide the dose reduction as only one day reduction of the average future time if anyone take the mammographic screening of breast cancer once a year over 30 years old. These alterations of apparatus produced the mammograms darkers, and the intensity of illumination of the view system brighter. These some improvements of the mammography have made the mammograms to be very high quality. As natural consequence mammographic diagnosis has some new roles. Screening of the breast cancer is the most important one. The diagnosis the lesion cancer or not is very important, too. If the lesion was found only with mammography, the biopsy under mammographic guidance is necessary. And the diagnosis of the extension of the cancer lesion is very important for surgical treatment. If the lesion has fine calcifications, mammography can diagnose the extent very objectively. PMID- 8709934 TI - [Ultrasonographic diagnosis of breast cancer with intraductal spreading of cancer cells]. AB - The estimation of intraductal lesions of the breast has become possible due to the careful examination of ultrasonic ductal images. The diagnostic accuracy ratio of the lesions was 94.4% by using our classification of dilated ducts by ultrasonography. The characteristics of the findings of intraductal spreading of breast cancer are that the wall of the duct is thick and uneven because of the proliferation of cancer cells, the tumor usually has a broad base all along the walls, and, when the duct is filed with cancer cells, the circumference becomes so varied that it appears outwardly irregular in size and shape. Microcalcifications and irregular nodules along the ductal network are also sometimes apparent. The diagnosis of the area of intraductal spreading of cancer cells could come about from examining the continuous spreading throughout the network by ultrasonography. We have succeedingly obtained successful results from using this method for the past eight years. The diagnostic accuracy ratio to determine the indication for breast conservative surgery and the extent of the breast excision was 94.2% in cases in which the tumor size was less than 3.0cm and the distance from the nipple was over 3.1cm, and in nonpalpable cases. In breast conservative surgery, the firm establishment of a proper diagnostic method is needed to prevent any cancerous tissue from being left. At this point in the evolution of the diagnostic method for intraductal spreading, ultrasonography is one of the key elements to achieve this goal. From now on, ultrasonography will be in the spotlight and take a more leading role in the continuing development of this diagnostic procedure. PMID- 8709935 TI - [Recent progress in the diagnosis of nonpalpable breast lesions]. AB - Recent advances in the diagnosis of nonpalpable breast lesions are reviewed. Recently, the technique of stereotaxic fine-needle breast biopsy for nonpalpable, mammographically detected breast lesions is available, and the technique has been proposed as an alternative to surgical biopsy. New equipment of stereo-guided breast biopsy system (LORAD co, system), which is produced for the use of stereotaxic core biopsy, is introduced. We examined 103 nonpalpable, mammographic lesions by this system and 14 carcinomas, including 11 noninfiltrating ones, were discovered. This system has been evident as a cost-effective and reliable method of evaluating mammographic lesions. PMID- 8709937 TI - [Progress of ductoscopy of the breast]. AB - Ductoscopy of the breast was initially performed utilizing a rigid scope. Improvements in endoscopic equipment and introduction of the silica-fiberscope have enabled observation of intraductal lesions. The silica-fiberscope can be gradually moved into peripheral sites, after which the endoscopic findings are compared with histological features and diagnostic imaging. The most common symptom dictating endoscopic examination is a discharge from the nipple in which no palpable mass is detected. The fiberscope is introduced through the duct orifice on the surface of the nipple. The characteristic feature of a cancerous lesion is a superficial spread along the surface of the ductal lumen, which differs from the typical features of intraductal papilloma. Microsampling methods such as tube curette cytology should be used to obtain a definitive diagnosis. Endoscopic examination can provide valuable information which will be helpful in the decision process such as; surgical methods, nipple-preservation, and breast conservation. PMID- 8709936 TI - [MR imaging for breast cancer]. AB - The potential role of magnetic resonance (MR) imaging in the diagnosis of breast disease has been investigated since the late 1970s. Conventional MR imaging has not been reliable for diagnosis of breast masses, and so contrast MRI using Gd DTPA has been performed. In particular the gradient echo (GRE) technique combined with administration Gd-DTPA (i.e., dynamic MRI) improves detection and tissue characterization of breast masses. In recent studies, breast MR imaging has shown up to 100% sensitivity in the detection of breast cancer. On the other hand, its specificity varies between 37% and 98% because of the fact that may benign lesions are also enhanced with many contrast media, similar to breast cancer. To improve the specificity of MR imaging, MR-guiding localization and fine needle aspiration biopsy for breast masses will be necessary. Recently breast conserving therapy has been widely accepted in our country, so we deduce that MRI is useful method for detecting multifocal or multicentric breast cancer. PMID- 8709938 TI - [Interphase cytogenetics of the breast tumors with fluorescence in situ hybridization (FISH) on cytologic preparation--its practice and clinical applications]. AB - Fluorescence in situ hybridization (FISH) study with the chromosome specific probes is performed in the interphase nuclei of the routinely processed cytologic preparation of the breast tumors. Numerical aberrations on the chromosomes 1, 3, 11 or 17 were detected in more than 80% of the malignant tumors, but not in the benign tumors. Marked heterogeneity of the polysomies is noted in the malignant tumor cells. A few malignant cases revealed monosomy of chromosome 17. No apparent correlation between the numerical abnormalities and the histological features in malignant tumors is identified. These results suggest that the interphase cytogenetics with FISH for the breast tumors may be useful for differential diagnosis of malignancy. The practice and the clinical applications of the FISH study are discussed. PMID- 8709939 TI - [The importance of tumor angiogenesis as a prognostic indicator in primary breast cancer]. AB - Recently, the importance of tumor angiogenesis in the process of tumor growth, progression and metastasis in solid tumors has been widely accepted. The prognostic value of angiogenesis has been demonstrated in a variety of solid tumors including breast cancer. In this report, we reviewed recent studies investigating on the value of intratumoral microvessel density (MVD), assessed by a sermiquantitative immunohistochemical assay with using factor-VIII related antibody or anti CD-31 antibody, as a prognostic indicator in primary breast cancer patients. Studies using factor-VIII related antibody showed that the average MVD ranged from 67.3 to 84.0 counts per mm2 area. When used by anti CD-31 monoclonal antibody, the average MVD were 120.3 approximately 135 counts per mm2 area in the range. More than 8 clinical investigations have showed that MVD was a potent prognostic indicator for relapse free survival and/or overall survival in both node-negative and -positive patients. Two reports concluded no prognostic value of MVD, however the average MVD of these two studies significantly differed from other reports. Thus, at present, angiogenesis grade seems to provide an independent prognostic value when the MVD was properly assessed. With respect to the relationship with conventional prognostic indicators, several reports showed the tendency that increased MVD was correlated with younger age and increase of tumor size below 3 cm diameter, however, some reports failed to demonstrate the tendency, suggesting that these correlations are still in controversial. Biological markers including ER, p53 and c-erB2 showed no correlation with the MVD in many studies including our investigation. Only a significant correlation we found was that MVD was increased in tumors with the expression of vascular endothelial growth factor and platelet-derived endothelial cell growth factor, which are noted to be potent endothelial growth factor. Since the evaluation of tumor angiogenesis as a prognostic indicator is now widely investigated in a prospective study, MVD might be introduced to the category of the criteria for determining the schedule of postoperative adjuvant therapy of breast cancer. PMID- 8709940 TI - [Genetic alterations and DNA-based diagnosis in breast cancer]. AB - Human carcinomas are generally considered to develop through the accumulation of various genetic abnormalities. The major types of genetic alterations that are frequently observed in breast cancer are amplification of protooncogenes (MYC, ERBB2); mutation of TP53; and loss of heterozygosity on chromosomes 1, 3p, 8p, 11p, 13q, 17q, 17, and 22q. The latter may correspond to losses or inactivations of tumor suppressor genes. Recently, two major distinct breast susptibility genes were isolated, namely BRCA1 and BRCA2. We performed PCR-SSCP analysis to determine the role of the BRCA1 gene in Japanese breast cancer and investigated how multiple genetic alterations contribute to tumor development and/or progression in primary breast cancer, using a large number of tumor materials. PMID- 8709941 TI - [Molecular mechanisms of breast cancer progression]. AB - Breast carcinoma is considered to occur as non-invasive carcinoma (DCIS) and to progress to the stage of invasive cancer. In DCIS, multiple gene and chromosome alterations are shown to occur already, and the pattern of alterations are concordant between DCIS and invasive components in most of individual tumors. Therefore, unknown molecular alterations are considered to exist that are involved in tumor invasion. A lot of alterations are also shown to occur in breast cancer cells, e.g., overexpression of growth factor receptors, cyclins and the molecules regulating cell-cycle, and cell-adhesion molecular. Clarification of signal-transducing mechanism of these molecules in cells and alterations of the system in cancer cells will be important as molecular mechanism of proliferation and progression of breast cancer. It is expected that the therapeutic strategy targetted to the control of these molecular events is developed in future. PMID- 8709942 TI - A gene for maackiain detoxification from a dispensable chromosome of Nectria haematococca. AB - In Nectria haematococca the MAK1 gene product converts a chick-pea (Cicer arietinum) phytoalexin, maackiain, into a less toxic compound. The presence of MAK1 in this fungal pathogen is also correlated with high virulence on chick-pea. Previous genetic analysis suggested that MAK1 is located on a meiotically unstable, dispensable chromosome. The unstable nature of this chromosome facilitated MAK1 cloning by allowing us to identify a subset of genomic cosmid clones likely to contain MAK1. Truncated forms of the chromosome, generated during meiosis, were isolated from strains either able (Mak+) or unable (Mak-) to metabolize maackiain and used to probe a chromosome-specific cosmid library. Only clones that hybridized exclusively to the chromosome from the Mak+ strain were then screened for their ability to transform a Mak- isolate to the Mak+ phenotype. A 2.7 kb HindIII-PstI fragment was subcloned from a cosmid conferring MAK1 activity, and its nucleotide sequence determined. Because MAK1 transcription is not induced strongly by maackiain, a reverse transcriptase-polymerase chain reaction was required to detect MAK1 transcription in a Mak+ strain, and to isolate MAK1 cDNA fragments. Comparison of the genomic and cDNA sequences of MAK1 revealed the presence of three introns and an open reading frame encoding a protein 460 amino acids in length. Two diagnostic domains in its deduced amino acid sequence suggest MAK1 encodes a flavin-containing mono-oxygenase. MAK1 is the first gene encoding maackiain detoxification to be cloned, and is the second functional gene cloned from this dispensable chromosome. Southern analysis of genomic DNA from ascospore isolates containing MAK2, MAK3, and MAK4 indicated that MAK1 is not homologous to other known maackianin-detoxifying genes. PMID- 8709943 TI - Two overlapping SOS-boxes in ColE operons are responsible for the viability of cells harboring the Col plasmid. AB - In this study, oligonucleotide-directed site-specific mutagenesis was used to change the consensus sequences of the LexA binding motifs in either one of the two SOS-boxes of the ColE7 operon. The results indicated that both mutants produced larger amounts of colicin than cells harboring the wild-type ColE7 plasmid. This finding would imply that two biologically functional SOS boxes exist in the ColE7 operon. In the non-induced state, no lysis of cells harboring wild-type plasmids occurred at 37 degrees C, whereas, cells harboring recombinant plasmids containing either one of the mutated SOS boxes underwent lysis within 100 min under the same conditions. This result indicated that adaptation of two SOS boxes of the ColE operon would obviously tightly control the expression of ColE operons. In such a way that it may prevent excessive expression of the lysis (cel) gene, thus safeguard the host cells from being lysed in ordinary living conditions. PMID- 8709944 TI - The hapC gene of Aspergillus nidulans is involved in the expression of CCAAT containing promoters. AB - The 5' regulatory region of the amdS gene of Aspergillus nidulans, which encodes an acetamidase required for growth on acetamide as a carbon and nitrogen source, contains a CCAAT sequence which is required for setting the basal level of amdS expression. Mobility shift studies have identified a factor in A. nidulans nuclear extracts which binds to this CCAAT sequence. In Saccharomyces cerevisiae the HAP3 gene encodes one component of a multisubunit complex that binds CCAAT sequences. A search of the EMBL and SwissProt databases has revealed an A. nidulans sequence with significant homology to the HAP3 gene adjacent to the previously cloned regulatory gene amdR. Sequencing of the remainder of this region has confirmed the presence of a gene, designated hapC, with extensive homology to HAP3. The predicted amino acid sequence of HapC shows extensive identity to HAP3 in the central conserved domain, but shows little conservation in the flanking sequences. A haploid carrying a hapC deletion has been created and is viable, but grows poorly on all media tested. This null mutant grows especially slowly on acetamide as a sole carbon and nitrogen source, indicating that hapC plays a role in amdS expression. In agreement with this notion, it has been shown that the hapC deletion results in reduced levels of expression of an amdS::lacZ reporter gene and this effect is particularly evident under conditions of carbon limitation. Nuclear extracts prepared from the hapC deletion mutant show no CCAAT binding activity to the amdS or gatA promoters, indicating that hapC may encode a component of the complex binding at this sequence. PMID- 8709945 TI - The C-terminal DNA-binding domain of Chironomus BR gene products shows preferential affinity for (dA.dT)-rich sequences. AB - Balbiani ring genes (BRs), the most active loci in the polytene chromosomes of the salivary gland of the midge Chironomus (Diptera), code for secretory giant peptides (the sp-I family). Evidence previously reported indicated that the conserved C-terminal region of proteins of the sp-I family had DNA-binding properties (assayed with sp-Ia), and one such region, derived from BR2.2, which codes for the product sp-Ib, might occur as a stable independent peptide, being transferred to the nucleus where it is detectable in the large BRs (BR1 and BR2), among other structures, by immunostaining. Here, we show that the C-terminal portion of one of the BR gene products, expressed as a glutathione-S-transferase fusion protein shows preferential affinity for A.T-rich sequences and binds with varying affinity to restriction fragments of the A.T-rich BR1 promoter. The binding was inhibited by distamycin, suggesting that the interaction involves the minor groove of the DNA. Analysis of the promoter fragments by gel electrophoresis indicated that most appeared to present a conspicuous bend, as deduced from their anomalous electrophoretic mobilities. Furthermore, the affinity of the C-terminal domain for the different promoter fragments appeared to correlate with the degree of bending. Thus, the C-terminal domain might play a role in controlling gene expression by binding to A.T-rich sequences, including those of the BR genes. PMID- 8709946 TI - Binding sites for maize nuclear proteins in the subterminal regions of the transposable element Activator. AB - Genetic data suggest that transposition of the maize element Activator (Ac) is modulated by host factors. Using gel retardation and DNase I protection assays we identified maize proteins which bind to seven subterminal sites in both ends of Ac. Four DNase I-protected sites contain a GGTAAA sequence, the other three include either GATAAA or GTTAAA. The specificity of the maize protein binding to Ac was verified by using a synthetic fragment containing four GGTAAA motifs as probe and competitor in gel retardation assays. All seven binding sites are located within regions required in cis for transposition. A maize protein binding site with the same sequence has previously been identified in the terminal inverted repeats of the maize Mutator element. Thus, the protein, that recognizes this sequence is a good candidate for a regulatory host factor for Ac transposition. PMID- 8709947 TI - Binding of Nicotiana nuclear proteins to the subterminal regions of the Ac transposable element. AB - Specific binding of Nicotiana nuclear protein(s) to subterminal regions of the Ac transposable element was detected using gel mobility shift assays. A sequence motif (GGTAAA) repeated in both terminal regions of Ac, was identified as the protein binding site. Mutation of two nucleotides in this motif was sufficient to abolish binding. Based on a series of competition assays, it is deduced that there is cooperative binding between two repeats, each similar to the GGTAAA motif. The binding protein is probably similar to a previously characterized maize protein which binds to a GGTAAA-containing motif located in the ends of Mutator. Moreover, we show that DNA from Ds1 competes for protein binding to Ac termini, and we show, by sequence analysis, that GGTAAA binding sites are present in the terminal region of Tgm1, Tpn1, En/Spm, Tam3, and Ds1-like elements. This suggests that the binding protein(s) might be involved in the transposition process. PMID- 8709948 TI - The Aspergillus nidulans genes chsA and chsD encode chitin synthases which have redundant functions in conidia formation. AB - We previously isolated three chitin synthase genes (chsA, chsB, and chsC) from Aspergillus nidulans. In the present work, we describe the isolation and characterization of another chitin synthase gene, named chsD, from A. nidulans. Its deduced amino acid sequence shows 56.7% and 55.9% amino acid identity, respectively, with Cal1 of Saccharomyces cerevisiae and Chs3 of Candida albicans. Disruption of chsD caused no defect in cell growth or morphology during the asexual cycle and caused no decrease in chitin content in hyphae. However, double disruption of chsA and chsD caused a remarkable decrease in the efficiency of conidia formation, while double disruption of chsC and chsD caused no defect. Thus it appears that chsA and chsD serve redundant functions in conidia formation. PMID- 8709949 TI - The glucose repressor gene cre1 of Trichoderma: isolation and expression of a full-length and a truncated mutant form. AB - The cre1 genes of the filamentous fungi Trichoderma reesei and T. harzianum were isolated and characterized. The deduced CREI proteins are 46% identical to the product of the glucose repressor gene creA of Aspergillus nidulans, encoding a DNA-binding protein with zinc fingers of the C2H2 type. The cre1 promoters contain several sequence elements that are identical to the previously identified binding sites for A. nidulans CREA. Steady-state mRNA levels for cre1 of the T. reesei strain QM9414 varied depending on the carbon source, being low on glucose containing media. These observations suggest that cre1 expression may be autoregulated. The T. reesei strain Rut-C30, a hyper-producer of cellulolytic enzymes, was found to express a truncated form of the cre1 gene (cre1-1) with an ORF corresponding to a protein of 95 amino acids with only one zinc finger. Unlike QM9414 the strain Rut-C30 produced cellulase mRNAs on glucose-containing medium and transformation of the full-length cre1 gene into this strain caused glucose repression of cbh1 expression, demonstrating that cre1 regulates cellulase expression. PMID- 8709950 TI - Isolation and characterization of two new negative regulatory mutants for nitrate assimilation in Chlamydomonas reinhardtii obtained by insertional mutagenesis. AB - Plasmid DNA carrying either the nitrate reductase (NR) gene or the argininosuccinate lyase gene as selectable markers and the corresponding Chlamydomonas reinhardtii mutants as recipient strains have been used to isolate regulatory mutants for nitrate assimilation by insertional mutagenesis. Identification of putative regulatory mutants was based on their chlorate sensitivity in the presence of ammonium. Among 8975 transformants, two mutants, N1 and T1, were obtained. Genetic characterization of these mutants indicated that they carry recessive mutations at two different loci, named Nrg1 and Nrg2. The mutation in N1 was shown to be linked to the plasmid insertion. Two copies of the nitrate reductase plasmid, one of them truncated, were inserted in the N1 genome in inverse orientation. In addition to the chlorate sensitivity phenotype in the presence of ammonium, these mutants expressed NR, nitrite reductase and nitrate transport activities in ammonium-nitrate media. Kinetic constants for ammonium (I4C-methylammonium) transport, as well as enzymatic activities related to the ammonium-regulated metabolic pathway for xanthine utilization, were not affected in these strains. The data strongly suggest that Nrg1 and Nrg2 are regulatory genes which specifically mediate the negative control exerted by ammonium on the nitrate assimilation pathway in C. reinhardtii. PMID- 8709951 TI - Variation in the ratio of physical to genetic distance in intervals adjacent to the Mla locus on barley chromosome 1H. AB - Variants of the pulsed-field gel electrophoresis technique were used in conjunction with two-dimensional DNA gel electrophoresis (2-DDGE) to determine the ratio of physical to genetic distance in two genetically defined intervals on barley chromosome 1H.2-DDGE analysis demonstrated that two loci that define a 0.3 cM interval, as determined by hybridization with BCD249, reside on a single 450 kb MluI fragment. This result indicates a maximum ratio of physical to genetic distance in this interval of 1500 kb/cM as compared to 3.7-4.2 Mb/cM for the barley genome as a whole. High molecular weight (HMW) DNA restricted with NotI and probed sequentially with MWG068 and BCD249 yield diffuse bands at approximately 2.8 Mb and 3.0 Mb in the C.I. 16151 and C.I. 16155 parental lines, respectively. These results suggest the maximum ratio of physical to genetic distance in the interval defined by these probes is 7.8 Mb/cM. Unique HMW DNA restriction fragment length polymorphisms (RFLP) were attributed to the presence of recombination breakpoints. Data from the recombination breakpoint analysis were used to estimate a ratio of physical to genetic distance of 2.5 Mb/cM in the Xbcd249.2-Xmwg068 interval and 0.465 Mb/cM in the Xbcd249.1-Xbcd249.2 interval. Both physical linkage and recombination breakpoint analysis indicate the Xbcd249.1-Xbcd249.2 interval is approximately five-fold smaller, physically, than the Xbcd249.2-Xmwg068 interval. PMID- 8709952 TI - The genetics of the repair of 5-azacytidine-mediated DNA damage in the fission yeast Schizosaccharomyces pombe. AB - We have recently demonstrated that Schizosaccharomyces pombe cells treated with the nucleoside analogue 5-azacytidine (5-azaC) require previously characterised G2 checkpoint mechanisms for survival. Here we present a survey of known DNA repair mutations which defines those genes required for survival in the presence of 5-azaC. Using a combination of single-mutant and epistasis analyses we find that the excision, mismatch and recombinational repair pathways are all required in some degree for the repair of 5-azaC-mediated DNA damage. There are distinct differences in the epistatic interactions of several of the repair mutations with respect to 5-azaC-mediated DNA damage relative to UV-mediated DNA damage. PMID- 8709953 TI - Analysis of the mutagenic properties of the UmuDC, MucAB and RumAB proteins, using a site-specific abasic lesion. AB - The mucAB and rumAB loci have been shown to promote mutagenesis to a greater extent than the structurally and functionally homologous Escherichia coli umuDC operon. We have analyzed the basis of this enhanced mutagenesis by comparing the influence of these operons, relative to umuDC, on the mutagenic properties of each of two abasic sites, specifically located in a single-stranded vector. Experiments with these vectors are useful analytical tools because they provide independent estimates of the efficiency of translesion synthesis and of the relative frequencies of each type of nucleotide insertion or other kind of mutagenic event. The umuDC, mucAB, and rumAB genes were expressed from their natural LexA-regulated promoter on low-copy-number plasmids in isogenic strains carrying a umuDC deletion. In addition, plasmids expressing the UmuD'C, MucA'B, or RumA'B proteins were also used. Compared to umuDC, the chief effect of mucAB was to increase the efficiency of translesion synthesis past the abasic site. The enhanced capacity of mucAB for translesion synthesis depended about equally on an inherently greater capacity to promote this process and on a greater susceptibility of the MucA protein to proteolytic processing. The RumA protein also appeared to be more susceptible to proteolytic processing, but the inherent capacity of the Rum products for translesion synthesis was no greater than that of UmuDC. dAMP was inserted opposite one of the two abasic sites studied at a somewhat greater frequency in strains expressing rum (82%) compared to those expressing umu (72%), which might result in higher mutation frequencies in rumAB than in umuDC strains. PMID- 8709954 TI - Molecular cloning and sequence analysis of a cDNA encoding a cysteine proteinase inhibitor from Sorghum bicolor seedlings. AB - A 711-bp cDNA encoding a cysteine proteinase inhibitor (cystatin) was isolated from a cDNA library prepared from 7-10 cm Sorghum bicolor seedlings. The nearly full-length cDNA clone encodes 130 amino acid residues, which include the Gin-Val Val-Ala-Gly motif, conserved among most of the known cystatins as a probable binding site for cysteine proteinases. The amino acid sequence of sorghum cystatin deduced from the cDNA clone shows significantly homology to those of other plant cystatins. The sorghum cystatin expressed in E. coli showed a strong papain-inhibitory activity. PMID- 8709955 TI - Gene conversion as a focusing mechanism for correlated mutations: a hypothesis. AB - Ways of producing complex mutational events without substantially raising the primary mutation rate are explored. If the small amount of DNA that is resynthesised through the action of the mismatch DNA repair system is not subject to further repair, the incidence of double mutations can increase by a factor of 100, while single mutations would increase by only 30%. Such a boost in the incidence of double mutations seems insufficient to meet the needs of higher organisms. For them, an alternative strategy would be to produce complex events by a succession of single mutations occurring in a correlated manner over several sexual generations. It is proposed that gene conversion may fulfill this role. Assuming that the resynthesis of DNA that occurs during gene conversion produces mutations in the conversion tract, one predicts a tendency for close mutations in corresponding sequences in the two homologous chromosomes, to promote, during conversion, further mutations in their vicinity. Semiquantitative calculations suggest that such a mechanism can be quite effective, provided the divergence between two paired chromosomes is around 10(-4) or less. Such a mechanism might constitute an adaptive mutation strategy acting at the population level. PMID- 8709956 TI - Use of a non-selective transformation technique to construct a multiply restriction/modification-deficient mutant of Neisseria gonorrhoeae. AB - A technique that allows for easy identification of transformants of Neisseria gonorrhoeae in the absence of selective pressure has been developed. A suicide vector that contains a gonococcal DNA uptake sequence was constructed to aid in DNA uptake. In this transformation procedure, a limiting number of cells is incubated with an excess amount of DNA, and the mixture is plated onto a non selective medium. At least 20% of the resulting colonies contained cells that had been transformed. This strategy was utilized to construct specific deletions of the S.N goI, II, IV, V and VII restriction-modification (R/M) genes. All five deletions were successfully incorporated into the chromosome of FA19, producing strain JUG029. Strain JUG029 could be transformed with non-methylated plasmid DNA while strain FA19 could not be transformed with such DNA. The development of a simple, non-selective transformation technique, coupled with the construction of a strain that is more permissive for DNA-mediated transformation, will aid in genetic manipulations of the gonococcus. PMID- 8709957 TI - A ribosomal protein gene (rpl32) from tobacco chloroplast DNA is transcribed from alternative promoters: similarities in promoter region organization in plastid housekeeping genes. AB - Multiple transcriptional start sites have been identified in the tobacco plastid ribosomal protein gene rpl32 by RNA mapping and in vitro capping techniques. A promotor with a canonical -10 Pribnow Box (P1) produces a major transcript in leaf chloroplasts. Transcription is also driven from additional promoters in non photosynthetic plastids from heterotrophically cultured cells (BY2 line). Among them, a second promoter located downstream (P2) generates the most prominent transcript in this type of cell. The absence of typical plastid promoter motifs upstream of this site and the higher steady-state level of the P2-derived transcript in BY2 cells suggest a distinct modulation of transcription. Mobility shift experiments also seem to indicate the existence of differences in protein DNA binding between both kinds of plastids with respect to a DNA fragment including the sequence upstream from the P2 starting site. The structure of the rpl32 promoter region is discussed in relation to that of other plastid housekeeping genes encoding elements of the genetic machinery. PMID- 8709958 TI - Telomere-homologous sequences occur near the centromeres of many tomato chromosomes. AB - Several bacteriophage lambda clones containing interstitial telomere repeats (ITR) were isolated from a library of tomato genomic DNA by plaque hybridization with the cloned Arabidopsis thaliana telomere repeat. Restriction fragments lacking highly repetitive DNA were identified and used as probes to map 14 of the 20 lambda clones. All of these markers mapped near the centromere on eight of the twelve tomato chromosomes. The exact centromere location of chromosomes 7 and 9 has recently been determined, and all ITR clones that localize to these two chromosomes map to the marker clusters known to contain the centromere. High resolution mapping of one of these markers showed cosegregation of the telomere repeat with the marker cluster closest to the centromere in over 9,000 meiotic products. We propose that the map location of interstitial telomere clones may reflect specific sequence interchanges between telomeric and centromeric regions and may provide an expedient means of localizing centromere positions. PMID- 8709959 TI - Characterisation of a new allele of pale cress and its role in greening in Arabidopsis thaliana. AB - A recessive mutant with white leaves was identified in a screen of a population of T-DNA-tagged Arabidopsis thaliana plants. The mutation is lethal, but plants develop almost to maturity under sterile conditions. The white areas in leaves are devoid of developed chloroplasts, but the plants frequently develop green sectors which contain green chloroplasts. Molecular characterisation of the affected gene revealed that the mutant is allelic to pale cress (pac), a recently described mutation, and was therefore named pac-2. Sequencing of cDNAs and the genomic region revealed several noteworthy features of this genetic locus. In pac 2 the T-DNA had inserted in the region of the promoter and abolished transcription of the PAC gene completely. Cytokinin induced greening in mature, white homozygous pac-2 plants, and therefore is likely to be responsible for the greening observed in callus and shoots induced on roots from such plants. However, the PAC transcript was found to be absent in both white leaves and green callus. Thus, since cytokinin induced greening in the absence of PAC RNA this plant hormone appears to be able to bypass PAC function. PMID- 8709960 TI - Regulation of acid phosphatases in an Aspergillus niger pacC disruption strain. AB - An Aspergillus niger strain has been constructed in which the pH-dependent regulatory gene, pacC, was disrupted. The pacC gene of A. niger, like that of A. nidulans, is involved in the regulation of acid phosphatase expression. Disruptants were identified by a reduction in acid phosphatase staining of colonies. Southern analysis demonstrated integration of the disruption plasmid at the pacC locus and Northern analysis showed that the disruption strain produced a truncated pacC mRNA of 2.2 kb (as compared to 2.8 kb in the wild type). The strain carrying the pacC disruption was used to assign the pacC gene to linkage group IV; this was confirmed by CHEF electrophoresis and Southern analysis. This strain further allowed us to determine which extracellular enzyme and transport systems are under the control of pacC in A. niger. Expression of the A. niger pacC wild-type gene and the truncated pacC gene showed that, in contrast to the auto-regulated wild-type expression, which was elevated only at alkaline pH, the truncated pacC gene was deregulated, as high-level expression occurred regardless of the pH of the culture medium. Analysis of the phosphatase spectrum by isoelectric focussing and enzyme activity staining both in the wild-type and the pacC disruptant showed that at least three acid phosphatases are regulated by the pacC. For the single alkaline phosphatase no pH regulation was observed. PMID- 8709961 TI - Identification of genetic factors for Alachlor tolerance in maize by molecular markers analysis. AB - Genetic factors controlling tolerance to the herbicide Alachlor in maize were localised by means of two different strategies. In the first approach, backcross (BC) plants, derived from pollen which had been subjected to selective pressure for resistance to the herbicide, were analysed for segregation distortion at 47 RFLP loci and compared to BC plants obtained from non-selected pollen. Preferential transmission of five chromosomal regions where putative QTLs (Quantitative Trait Loci) are localised was revealed in the BC plants from selected pollen. A second approach was based on a classical linkage analysis for segregation of the same set of RFLPs and factors controlling the trait, in a BC population of 210 individuals, by means of regression analysis. This study detected seven significant loci in four genomic regions. Overall, two loci revealed both segregation distortion and association with the expression of the trait, indicating linkage to genes expressed in both gametophytic and sporophytic phase. Three chromosomal regions appeared to carry factors involved in plant tolerance to Alachlor which are not expressed in pollen. Conversely, three loci were linked to factors selectable in pollen, but did not reveal significant association with tolerance in the plant in the segregating populations. PMID- 8709962 TI - cAMP inhibits bud growth in a yeast strain compromised for Ca2+ influx into the Golgi. AB - Biochemical and physiological studies have implicated cAMP and cAMP-dependent protein kinase (PKA) in a plethora of essential cellular processes. Here we show that yeast cells partially depleted of PKA activity (due to a tpkw mutation) and bearing a lesion in a Golgi-localized Ca2+ pump (Pmr1), arrest division with a small bud. The bud morphology of the arrested tpk1w pmr1 mutant cells is characteristic of cells in S phase; however, the terminal phenotype of processes such as DNA replication and nuclear division suggests arrest at the G2/M boundary. This small bud, G2-arrest phenotype is similar to that of strains with a defect in cell wall biosynthesis (pkc1) or membrane biogenesis (och1); however, the biochemical defect may be different since the tpk1w pmr1 double mutants retain viability. The growth defect of the tpk1w pmr1 mutant can be alleviated by preventing the increase in cellular cAMP levels that is known to be associated with a decrease in PKA activity, or by supplementing the medium with millimolar amounts of Ca2+. Although the biochemical consequences of this increase in cAMP concentration are not known, the small-bud phenotype of the double mutant and the known protein processing defect of the pmr1 lesion suggest that the localization or function of some membrane component might be compromised and susceptible to perturbations in cellular cAMP levels. One candidate for such a protein is the cAMP-binding membrane ectoprotein recently described in yeast. PMID- 8709963 TI - A kinase-encoding gene from Colletotrichum trifolii complements a colonial growth mutant of Neurospora crassa. AB - Colletotrichum trifolii is a fungal pathogen which is responsible for anthracnose disease of alfalfa. To initiate research on molecular communication in this fungus, a kinase-encoding gene (TB3) and the corresponding cDNA were cloned and sequenced. The deduced amino acid sequence of TB3 closely resembles that of a Neurospora crassa serine/threonine protein kinase, COT1, required for hyphal elongation and branching. The C-terminal catalytic domains of TB3 and COT1 are highly conserved but the N-terminal regions are divergent, particularly in the homopolymeric glutamine repeats of TB3. Northern analysis indicated that TB3 expression was highest 1 h after inducing conidial germination and 1 h before germ tubes were first observed. Expression of TB3 transcripts returned to constitutive levels by 4 h after induction of germination. TB3 complemented the cot-I mutant of Neurospora crassa, demonstrating the functional conservation of this kinase between a pathogenic and a saprophytic fungus. PMID- 8709964 TI - Induction of the Escherichia coli UVM response by oxidative stress. AB - UVM (ultraviolet modulation of mutagenesis) is a recently described recA independent, inducible mutagenic phenomenon in which prior UV irradiation of Escherichia coli cells strongly enhances mutation fixation at a site-specific 3 N4-ethenocytosine (epsilon C) lesion borne on a transfected single-stranded M13 DNA vector. Subsequent studies demonstrated that UVM is also induced by alkylating agents, and is distinct from both the SOS response and the adaptive response to alkylation damage. Because of the increasing significance being attributed to oxidative DNA damage, it is interesting to ask whether this class of DNA damage can also induce UVM. By transfecting M13 vector DNA bearing a site specific epsilon C lesion into cells pretreated with inducing agents, we show here that the oxidative agent H2O2 is a potent inducer of UVM, and that the induction of UVM by H2O2 does not require oxyR-regulated gene expression. UVM induction by H2O2 appears to be mediated by DNA damage, as indicated by the observation of a concomitant reduction in cellular toxicity and UVM response in OxyRc cells. Available evidence suggests that UVM represents a generalized cellular response to a broad range of chemical and physical genotoxicants, and that DNA damage constitutes the most likely signal for its induction. PMID- 8709965 TI - A putative membrane protein, Pho88p, involved in inorganic phosphate transport in Saccharomyces cerevisiae. AB - Transcription of a regulatory gene, PHO81, in the phosphatase regulon of Saccharomyces cerevisiae is repressed by inorganic phosphate (Pi) in the medium via that same regulatory system. The activity of Pho81p, the product of PHO81, is also inhibited by a high concentration of Pi in the medium. Increased dosage of PHO86, a gene encoding a putative membrane protein associated with a Pi transporter complex, activates the Pi-inhibited Pho81p produced under the control of the GAL1 promoter. A new gene, PHO88/ YBR106w, has now been identified as a multicopy suppressor of the rAPase- phenotype of the cells caused by the Pi inhibition of Pho81p. The pho86 disruptant expressed rAPase activity in high-Pi medium, while the pho88 disruptant did not. The delta pho86 delta pho88 double disruption resulted in enhanced synthesis of rAPase under the high-Pi condition and conferred arsenate resistance on the cells than those in single disruptants of these genes. Its hydropathy profile and the results of an analysis of its cellular localization suggested that Pho88p is a membrane protein similar to Pho86p. Both disruption and high dosage of PHO88 or PHO86 resulted in reduced Pi uptake. These findings suggest that Pho88p is also involved in Pi transport and modulates Pho81p function together with Pho86p. PMID- 8709966 TI - Regulation of the ribA gene encoding GTP cyclohydrolase II by the soxRS locus in Escherichia coli. AB - We isolated a promoter that is inducible by paraquat, a superoxide-generating agent, from Escherichia coli using the promoter-probe plasmid pRS415. Sequence analysis revealed that the promoter derives from the ribA gene encoding GTP cyclohydrolase II, which is the first enzyme in the biosynthetic pathway of riboflavin. We fused the lacZ gene with the ribA promoter to monitor the expression of the gene in the single-copy state. LacZ expression from the ribA promoter was induced about eight-fold by 200 microM paraquat. Other known superoxide generators, menadione and plumbagin, also induced the expression of beta-galactosidase in the fusion strain. On the other hand, no significant induction was observed following treatment with hydrogen peroxide, ethanol or heat shock. Induction of beta-galactosidase was significantly reduced by the introduction of a delta sox-8::cat or soxS3::Tn10 mutation into the fusion strain, indicating that the ribA gene is a member of the soxRS regulon. The transcriptional start site was determined by primer extension analysis and putative binding sites for SoxS in both orientations were identified. GTP cyclohydrolase II activity in soluble extracts of E. coli increased more than three-fold on treatment with paraquat. This increase was dependent on the soxRS locus, and reflects the increase in transcript levels. However, flavin pools did not change significantly. A possible role for ribA induction during superoxide stress is discussed. PMID- 8709967 TI - Phosphorylation/dephosphorylation steps are key events in the phytochrome mediated enhancement of nitrate reductase mRNA levels and enzyme activity in maize. AB - We provide evidence to show that the increase in nitrate reductase (NR) transcript level stimulated by red light is mediated via a phosphorylation dependent step. The light-stimulated enhancement of NR transcript level was significantly inhibited by H-7, a protein kinase inhibitor, whereas okadaic acid (OKA), a phosphatase inhibitor, had no effect. Phorbol myristate acetate (PMA), an activator of protein kinase C (PKC) enhanced the NR transcript level in dark grown leaves. No correlation between changes in NR transcript level and NR activity (NRA) was observed. Inhibition of NRA by OKA and stimulation by H-7 indicated that NRA is increased by dephosphorylating the enzyme. We have identified a protein kinase (C type) that can phosphorylate the purified NR in vitro without the involvement of other accessory proteins. By in vivo labelling with 32P and immunoprecipitation of NR with NR antibodies it was found that in the presence of OKA most NR protein (NRP) was present in phosphorylated state, while with H-7 the reverse was seen. The red (R) and far-red (FR) light reversible experiments suggested that phytochrome (Pfr, an active form) stimulation of NRA is mediated by dephosphorylation of the enzyme, suggesting that Pfr regulates both NR transcription and NRA via phosphorylation/dephosphorylation steps controlled by separate signal transduction pathways. PMID- 8709968 TI - Sequence variation in the putative replication origins of the five genetically distinct basic Euoenothera plastid chromosomes (plastomes). AB - Approximately 4,200 nucleotides of the 16S/23S rDNA spacer and the 5' region flanking the rrn operon of the plastid chromosomes representing the five basic, phylogenetically related Euoenothera plastomes were sequenced and compared. The sequences that harbor the putative replication origins are almost identical except for a 785-bp intercistronic segment between the genes for the 16S rRNA and trnI. Differences are mainly caused by insertions/deletions and duplications; the predicted potential for formation of quite extensive secondary structure differs among the plastomes. Unexpected intraplastome variation has also been noted. Furthermore, the sequence-based and published genetically deduced plastome pedigrees differ significantly. PMID- 8709969 TI - Genetic evidence for functional specificity of the yeast GCN2 kinase. AB - In yeast the GCN2 kinase mediates translational control of GCN4 by phosphorylating the alpha subunit of eIF-2 in response to extracellular amino acid limitation. Although phosphorylation of eIF-2 alpha has been shown to inhibit global protein synthesis, amino acid starvation results in a specific activation effect on GCN4 mRNA translation. Under the same conditions, translation of other mRNAs appears only slightly affected. The mechanism responsible for the observed selectivity of the GCN2 kinase is not clear. Here, we present genetic evidence that suggests that locally restricted action of the GCN2 kinase facilitates GCN4-specific translational regulation. PMID- 8709970 TI - Monophosphoryl lipid A induces pharmacologic 'preconditioning' in rabbit hearts without concomitant expression of 70-kDa heat shock protein. AB - The purpose of this study was to evaluate the protective effect of a new endotoxin analogue, monophosphoryl lipid A (MLA) in a rabbit model of myocardial ischemia/reperfusion and to show if this protection was mediated via synthesis of 70 kDa heat shock protein (HSP 70). Three groups of New Zealand White rabbits underwent 30 min coronary occlusion, followed by 4 hours reperfusion. First group of rabbits (n = 6) were treated with 0.35 ml vehicle (40 % propylene glycol, 10 % ethanol in water). The second and third group of rabbits (n = 6-8) were treated with MLA (35 micrograms/kg, i.v.) 12 and 24 hours prior to ischemia and reperfusion. MLA treatment either 12 or 24 h prior to ischemia/reperfusion demonstrated significantly reduced infarct size (12.5 +/- 1.7 and 14.7 +/- 2.1% for 12 and 24 h) when compared with vehicle control (40.4 +/- 8.6%, mean +/- S.E.M, p < 0.05). No significant differences in the infarct size was observed between the 12 and 24 h MLA treated groups. The area at risk was not significantly different between the three groups. Baseline values of heart rate, systolic and diastolic blood pressure were not significantly different between the control and MLA treated groups. However, the systolic as well as diastolic blood pressure during reperfusion were significantly lower in rabbits treated with MLA. Western blot analysis of the protein extracts of the hearts (n = 2/group) demonstrated no increase in the expression of the inducible form of HSP 70 following treatment with MLA. We conclude that MLA has significant anti infarct effect in rabbit which is not mediated by the cardioprotective protein HSP 70. The anti-infarct effect of this drug is superior to the reported protective effects of delayed ischemic or heat stress preconditioning. We hypothesize that the pharmacologic preconditioning afforded by MLA is accomplished via a unique pathway that bypasses the usual intracellular signaling pathways which lead to the myocardial protection with the expression of heat shock proteins. PMID- 8709971 TI - Effect of acute exercise on glutathione deficient heart. AB - The role of glutathione (GSH) in myocardial antioxidant defense was investigated in Swiss-Webster mice either performing swim exercise to exhaustion or rested in both the GSH adequate (GSH-A) and GSH deficient (GSH-D) states. GSH deficiency was accomplished by injecting mice with L-buthionine [S,R]sulfoximine (BSO; 2 nmol/kg body wt, i.p.) and providing BSO (20 mM) in drinking water for 12 days. GSH and glutathione disulfide (GSSG) contents in the GSH-D hearts were decreased to 10 and 8%, respectively, of those in the GSH-A mice. This decrease was associated with a significant decline of the total glutathione level in the liver, skeletal muscle and plasma. Myocardial GSH peroxidase and GSH sulfur transferase activities decreased significantly following GSH deficiency, whereas superoxide dismutase activity was significantly elevated. GSH deficiency did not affect exercise endurance performance. However, exhaustive exercise decreased GSH content in the myocardium of the GSH-A and GSH-D mice by 22 and 44% (p < 0.05), respectively. The GSH:GSSG ratio was not altered significantly following exercise because of a concomitant decrease in GSSG (p < 0.05). gamma Glutamyltranspeptidase activity was significantly increased after exercise, especially in the GSH-D hearts (72%; p < 0.05). GSH content after exercise correlated negatively with exercise time in both GSH-A and GSH-D mice (p < 0.05). These data indicate that GSH is actively used in the myocardium during prolonged exercise at moderate intensity and that GSH deficiency is tolerated by the heart, possibly compensated for by an increased GSH uptake from the plasma. PMID- 8709972 TI - Molecular basis of pathophysiology of Indian childhood cirrhosis: role of nuclear copper accumulation in liver. AB - Indian Childhood Cirrhosis (ICC) is a disease of abnormal copper metabolism commonly characterized by swelling and degeneration of liver cells along with the presence of orcein staining deposits of copper. Hepatic copper content of ICC patients was about 43 fold higher than those of control subjects. The data on sub cellular distribution of copper revealed massive accumulation of Copper (73%) of total cell copper) in the nuclear fraction (455 micrograms Cu/g tissue nuclei). On further distribution of copper in nuclear fraction, the enrichment of copper in heterochromatin and euchromatin of ICC nuclei was found to be 48 and 15 fold higher over control fractions respectively. The ultra-violet spectra of heterochromatin and euchromatin isolated from ICC nuclear fraction showed a broad absorption maxima as compared to controls. Further, A260/A280 ratio was markedly lower in heterochromatin and euchromatin of ICC liver in comparison to controls. An antioxidant enzyme, catalase activity was also significantly reduced in ICC liver as compared to control. Further, DNA fragmentation studies indicated that there was significantly increased DNA fragmentation in ICC liver. Collectively, these findings suggest that massive accumulation of copper in nucleus and decrease in catalase activity was associated with DNA fragmentation in hepatocyte of ICC disease. PMID- 8709973 TI - Influence of sodium pentosan polysulphate and certain inhibitors on calcium oxalate crystal growth. AB - Calcium oxalate crystal growth and aggregation leads to the formation of renal calculi. It is known to be inhibited by several compounds both in vitro and in vivo conditions. The present study highlights the inhibitory potential of sodium pentosan polysulphate (SPP), a semi-synthetic glycosaminoglycan (GAG) on calcium oxalate crystal growth in vitro. Its efficacy was compared with those of known inhibitors like pyrophosphate, heparin and chondroitin-4-sulphate. Of the above compounds pyrophosphate was found to be the most potent inhibitor. Among the GAGs, SPP exhibited 80% inhibitory activity as compared to heparin. A lesser degree of inhibition was observed with chondroitin-4-sulphate. PMID- 8709974 TI - Platelet calcium pump activity in essential hypertensives and their first-degree relatives. AB - Intracellular free Ca2+ concentration has been shown to be elevated in platelets of patients with essential hypertension. This study was designed to characterize Ca(2+)-pump activity of the platelet membranes (surface and intracellular) in these patients A double-blind study was carried out. Untreated and treated (on beta-blockers) essential hypertensives were studied in comparison with normotensive control subjects. First degree blood relatives of essential hypertensives were also studied. The Ca2+ activation kinetics of the enzyme showed a significant decrease in the Vmax (for the plasma- and intracellular membranes) and Km (for the intracellular membranes) in the essential hypertensive patients. Increased platelet membrane cholesterol content was observed in these patients. Lowered Ca2(+)-efflux by Ca2(+)-ATPase may lead to elevated intracellular free Ca2(+)-levels in platelet of essential hypertensives. A lowered Ca2(+) -ATPase activity may emerge as a marker for essential hypertension. PMID- 8709975 TI - Regulation of c-fos mRNA expression in Sertoli cells by cyclic AMP, calcium, and protein kinase C mediated pathways. AB - The role of second messenger pathways, cyclic AMP, calcium, and protein kinase C (PKC) in the transcriptional regulation of c-fos protooncogene expression in rat Sertoli cells was investigated. c-fos expression was monitored by Northern blot analysis. Although the action of FSH on Sertoli cells is considered to be mediated by cAMP, dibutyryl cAMP (dbcAMP), a potent membrane permeable analog of cAMP, induced much less c-fos mRNA expression than FSH ( < 50%) suggesting that additional cAMP-independent mechanisms may mediate the effect of FSH on c-fos. Specific intracellular inhibitors of PKC decreased c-fos induction in response to FSH by more than 50%. Ionomycin, which increases intracellular free calcium concentration, induced c-fos expression significantly. These data demonstrate that Sertoli cell c-fos mRNA expression is under multifactorial regulation by cAMP, calcium, and PKC. PMID- 8709976 TI - Chronic muscle stimulation increases lactate transport in rat skeletal muscle. AB - The aim of this study was to examine the effects of chronic low frequency stimulation on the lactate transport across the plasma membrane of the tibialis anterior (TA) muscle of the rat. Stimulating electrodes were implanted on either side of the peroneal nerve in one hindlimb. Chronic stimulation (10 Hz, 50 microsecond bursts, 24h/day) commenced 7 days after surgery, and were continued for 7 days. Animals were then left for 24 h, and thereafter muscles were obtained. Cytochrome C-oxidase activity was increased 1.9-fold in the stimulated TA compared to the control TA (p < 0.05). Lactate transport (zero-trans) was measured in giant sarcolemmal vesicles obtained from the chronically stimulated TA and the control TA. At each of the concentrations used in these studies a significant increase in lactate transport was observed; 2.8-fold increase at 1 mM lactate p < 0.05); 2-fold increases at both 30 mM and 50 mM lactate p < 0.05). These studies have shown that lactate transport capacity is markedly increased in response to chronic muscle contraction. PMID- 8709977 TI - Expression of a 24 kDa GTP-binding protein (Gn24) is increased in lovastatin treated human erythroleukemia cells. AB - A major 27 kDa particulate and a minor 24 kDa cytosolic GTP-binding protein was detected in HEL cells upon incubation with [alpha-32P]GTP of nitrocellulose blots containing polypeptides separated using SDS-PAGE. Addition of lovastatin (30 microM) to HEL cells in culture inhibited protein synthesis by approximately 35%. However, this treatment resulted in a 5-fold increase, as quantitated by [alpha 32P]GTP binding, in the amount of cytosolic 24 kDa GTP-binding protein. Addition of cycloheximide plus lovastatin to cells in culture abolished the observed increase in 24 kDa GTP-binding protein. Incubation of cells with lovastatin plus [R,S]-[5-(3)H] mevalonolactone resulted in the incorporation of radioactivity into several polypeptides in both the cytosolic and particulate fractions including a polypeptide of molecular mass of 24 kDa in the cytosol. The mobility of this 24 kDa isoprenylated protein on SDS-PAGE was identical to that of the GTP binding protein increased in response to lovastatin. However, the 24 kDa protein remained in the cytosol after undergoing isoprenylation. The 24 kDa protein was distinct from the HEL cell, G25K/CDC42Hs GTP-binding protein and the GTP-binding protein that was a substrate for botulinum toxin C3 catalyzed ADP-ribosylation. Results demonstrate that lovastatin specifically increases the expression of a 24 kDa GTP-binding protein in HEL cells and that, isoprenylation of low molecular mass GTP-binding protein(s) may have function(s) in addition to its role in the targetting of these proteins to cell membrane. PMID- 8709978 TI - Induction of early growth response-1 gene by interleukin-1 beta and tumor necrosis factor-alpha in normal human bone marrow stromal an osteoblastic cells: regulation by a protein kinase C inhibitor. AB - The early growth response-1 (Egr-1) gene has been identified as a nuclear transcriptional factor and implicated in the regulation of growth and differentiation of osteoblastic cells. In the present study, we investigated whether Egr-1 mRNA is expressed and induced by interleukin-1 beta, (IL-beta) and tumor necrosis factor-alpha (TNF-alpha) in normal human bone marrow stromal (HBMS) and osteoblastic (HOB) cells. Results demonstrate a very low basal expression of Egr-I mRNA which is induced by IL-1 beta, and TNF-alpha in a time- and dose-dependent manner. Egr-1 mRNA induction was detectable within 15 min, reached maximal by 60 min and thereafter declined to basal levels by 120 min. Induction of Egr-1 mRNA by IL-1 beta and TNF-alpha was completely inhibited by H 7 suggesting the mediation of protein kinase C. The induction by IL-1 beta and TNF-alpha of Egr-1 mRNA was independent of de novo protein synthesis since this induction was also observed in the presence of protein synthesis inhibitor cycloheximide. Fetal bovine serum and cycloheximide also independently induced the Egr-1 mRNA. Actinomycin D experiments demonstrated that Egr-1 mRNA is degraded very rapidly with a half-life of 30 min. Our results demonstrate the expression of Egr-1 gene and its induction by IL-1 beta, and TNF-alpha in normal human bone marrow stromal (osteoprogenitor) and osteoblastic cells in primary cultures. Data also reveal that the expression of Egr-1 gene is inhibited by protein kinase C inhibitor H-7 suggesting that the activation of protein kinase C or other protein kinases resulting in the phosphorylation of specific transcription factor(s) is the first immediate early step in the induction of immediate-early Egr-1 gene by IL-1 beta, and TNF-alpha. Results also suggest that Egr-1 is an important mediator of IL-1 beta and TNF-alpha action in normal human osteoblastic cells. PMID- 8709979 TI - Postischemic injury in isolated rat hearts is not aggravated by prior depletion of myocardial glutathione. AB - The aim of this study was to test the hypothesis that a decreased myocardial concentration of reduced glutathione (GSH) during ischemia renders the myocardium more susceptible to injury by reactive oxygen species generated during early reperfusion. To this end, rats were pretreated with L-buthionine-S,R-sulfoximine (2 mmol/kg), which depleted myocardial GSH by 55%. Isolated buffer-perfused hearts were subjected to 30 min of either hypothermic or normothermic no-flow ischemia followed by reperfusion. Prior depletion of myocardial GSH did not lead to oxidative stress during reperfusion, as myocardial concentration of glutathione disulfide (GSSG) was not increased after 5 and 30 min of reperfusion. In addition, prior depletion of GSH did not exacerbate myocardial enzyme release, nor did it impair the recoveries of tissue ATP, coronary flow rate and left ventricular developed pressure during reperfusion after either hypothermic or normothermic ischemia. Even administration of the prooxidant cumene hydroperoxide (20 microM) to postischemic GSH-depleted hearts during the first 10 min of reperfusion did not aggravate postischemic injury, although this prooxidant load induced oxidative stress, as indicated by an increased myocardial concentration of GSSG. These results do not support the hypothesis that a reduced myocardial concentration of GSH during ischemia increases the susceptibility to injury mediated by reactive oxygen species generated during reperfusion. Apparently, myocardial tissue possesses a large excess of GSH compared to the quantity of reactive oxygen species generated upon reperfusion. PMID- 8709980 TI - Release of ischemia in paced rat Langendorff hearts by supply of L-carnitine: role of endogenous long-chain acylcarnitine. AB - Rat Langendorff hearts perfused with media that do not contain erythrocytes or fluorocarbon as oxygen carriers are borderline aerobic during 5 Hz pacing. This follows from the release of catabolic products measured: lactate, urate and Iysophosphatidyl-choline (IysoPC). Addition of L-carnitine to the perfusion medium reduced the level of these compounds, while the release of long-chain acylcarnitine (LCAC) increased. Previously, we found (Biochim Biophys Acta 847:62 66,1985) that micromolar LCAC protects membranes during reperfusion after ischemia. Therefore, the observed inverse relation between LCAC and the other compounds measured suggests that LCAC is the basis of an acute relief of imminent ischemia by carnitine addition. LCAC may be released from various cell types, including vascular endothelium, as demonstrated. The cationic amphiphilic nature of LCAC is responsible for protection of membrane functions in imminent ischemia. PMID- 8709981 TI - Alteration of extracellular matrix in dilated cardiomyopathic hamster heart. AB - The purpose of this study was to characterize the collagen in hereditary dilated cardiomyopathic hamster hearts, and to examine the participation of the collagen in the occurrence and progression of cardiomyopathy. BIO 53.58 hamsters (5, 10, 20 weeks old) were used as the model of dilated cardiomyopathy. Flb hamsters were used as controls. The collagen content was almost constant at any age in the Flb hamsters, but increased with age in BIO 53.58 hamsters. Type III collagen increased significantly in BIO 53.58 hamsters at 10 weeks. The acetic acid solubility of collagen decreased in BIO 53.58 hamsters as the fibrosis progressed, but was unchanged in controls. Reducible crosslinks showed a tendency to decrease progressively in BIO 53.58 hamsters. There were no differences between Flb and BIO 53.58 hamsters at 5 weeks, but its expression in BIO 53.58 hamsters at 10 and 20 weeks of age increased compared to Flb controls. These findings indicate that in the early phase of cardiomyopathy the extracellular matrix of the myocardium is rich in type III collagen. In the later phase, the matrix resembles that of hard tissues, whose collagen is mainly of type I collagen and is insoluble. These data suggest that the increased collagen synthesis may impair the cardiac function in the development of cardiomyopathy. PMID- 8709982 TI - [Simultaneous determination of the toxicologically relevant elements: lead, cadmium, and nickel with graphite furnace atomic absorption spectrometry]. AB - A new generation of graphite furnace atomic absorption spectrometer enables the simultaneous determination of several trace elements. Using an optimized temperature-time-program for ashing and atomization, the Smith-Hieftje background correction and a matrix modifier, the determination of Pb, Cd and Ni in one run is possible. The detection limits of the new method of 0,039 mg Pb, 0,006 mg Cd and 0,022 mg Ni per kg sample, permit the determination of these elements in soils, foodstuffs and human food. These detection limits comply with the guidelines of the "Zentrale Erfassungs- und Bewertungsstelle fur Umweltchemikalien" (central registration- and assessment office for environmental chemicals) of the "Bundesgesundheitsamt" (german public health department). The new multielement method reduces measurement time and costs, i.e. graphite tubes, gas, matrix modifier, reference material and sample solution, in comparison to conventional single-element GF-AAS. PMID- 8709983 TI - Effectiveness of microwave based digestion procedures for the demineralization of human milk and infant formulas prior to fluorometric determination of selenium. AB - The use of two types of microwave oven, one domestic (A) and the other specially designed for laboratory (B) for microwave-based digestion, to destroy the organic matter in milk and infant formulas prior to fluorometric determination of selenium is studied in order to check their applicability and reliability. In both systems the best results were obtained using the acid mixture HNO3-H2O2, and additional treatment with perchloric acid (60%) in a Thermoblock. The application of two different microwave-based digestion procedures to infant formulas was free of interferences. In the case of human milk, it was necessary to use the standard addition method. Values obtained for detection and quantification limits, precision and accuracy of the fluorometric determination of selenium show the usefulness of both digestion procedures. The oven designed for laboratory use (B) is preferential as it permits simultaneous digestion of eight samples. PMID- 8709984 TI - Magnetoreception: why is conditioning so seldom successful? AB - Conditioning is a highly successful method for analyzing the sensory capacities of animals. With magnetic stimuli, however, it does not seem to work: negative results by far outnumber the positive ones. This is true for cardiac and operant conditioning as well as for directional training. The reasons for these failures are largely unclear. They may stem from the function of the magnetic field as orientation cue and from the fact that the magnetic field never undergoes a rapid change in nature, which means that animals might not be adapted to respond to such changes. Moreover, since the magnetic field contains directional information, animals might evade the problems arising from self-produced movements by calling on magnetic information only when needed for orientation. In view of this, conditioning does not appear to be a suitable technique for testing magnetic sensitivity. PMID- 8709987 TI - Managing transplant care: an HMO perspective. PMID- 8709985 TI - [Regulation of cell function by level of hydration]. AB - The hydration state of mammalian cells is dynamic and changes within minutes under the influence of nutrients, hormones and oxidative stress. Such changes in cell hydration act as an independent signal which modulates cellular metabolism and gene expression by activating intracellular signalling systems. Although the structures which sense hydration changes are unknown, this creates a novel and elegant mechanism for adaptation of cell function and gene expression to environmental challenges and provides new aspects for clinical medicine. PMID- 8709986 TI - Acetylation of alpha-tocopherol by the squash beetle, Epilachna borealis. Defense mechanisms of arthropods, 134 [1]. PMID- 8709988 TI - On realistic survival data, Neoral dosage. PMID- 8709989 TI - Finding dialysis care in the Ukraine: Ohio technician joins efforts to bring medical care to Russia. PMID- 8709990 TI - Mortality: is it fair to compare PD & HD? PMID- 8709991 TI - Optimizing hemodialysis access: a teaching tool. PMID- 8709992 TI - ESRD in the American Indian population. PMID- 8709993 TI - Managing transplant care: an HMO perspective. Part II: Selecting "centers of expertise". PMID- 8709995 TI - Satisfaction belongs in the health care equation. PMID- 8709994 TI - Organizing a living donor pool: could the concept work? PMID- 8709996 TI - Nursing and health care reform. PMID- 8709997 TI - Measuring patients' ability to read and comprehend: a first step in patient education. AB - Recent reports released by the U.S. Department of Education indicate that approximately 90 million adults have poor reading and comprehension skills. This information highlights the importance of considering the literacy level of clients when planning patient education programs. The purpose of this investigation was to measure, in urban public clinics, the discrepancy between a self-report of the highest grade completed in school and the actual reading and comprehension levels of patients, measured by the WRAT and CLOZE. Twenty-five subjects, 22 African Americans and 3 Caucasians, participated in this study. The findings indicated that, even though the self-reported highest grade completed showed a mean of twelth grade, the actual mean reading level was below eighth grade. The results on comprehension showed that 52% (N = 13) of the subjects needed additional instructions after reading materials and 24% were unable to comprehend what they read. The demographics of people with poor literacy skills suggests that they represent a significant proportion of health care consumers who visit health departments, primary care facilities, and community-based health centers. PMID- 8709998 TI - To ration or not to ration: that is the question. PMID- 8709999 TI - In search of a nursing case management model for rural hospitals. AB - Research has provided increasing evidence that nursing case management is a cost effective way to maximize health outcomes for high-risk, high-cost, high-volume patients (Ely, Walker, & Berger, 1993: Etheridge & Lamb, 1989; Krieger, Connell, & LoGerfo, 1992; Green, Lovely, Miller, & Ondrich, 1989; Schull, Tosch, & Wood, 1992; Zander, 1988). However, models of nursing case management developed in urban settings and tested primarily on urban populations do not address the needs of rural people and areas. This article proposes a definition of nursing case management for rural hospitals that is grounded in the reality of rural practice and offers a meaningful approach to the provision of care to underserved populations, cost containment, and recruitment and retention of registered nurses. PMID- 8710000 TI - The ethical dilemmas of mandatory prenatal and newborn HIV testing. PMID- 8710001 TI - Storytelling: a way of connecting. AB - Storytelling is a way of knowing, in nursing, that which we have taken for granted. It is a powerful way to communicate with students, patients, and colleagues. A well-told story has the capability of remaining vivid in memory long after facts and figures are forgotten. This essay describes how storytelling can facilitate the processes of nursing education, practice, and research. PMID- 8710002 TI - Recruiting future nurses: a collaborative project. AB - Society as a whole must be involved in making certain there are enough professionals in the future to provide care for the health needs of both rural and urban communities. As a result of this need. Area Health Education Centers (AHECs) across North Carolina have expanded their health care career awareness efforts. One AHEC focused on the concept of awareness of nursing careers by the community, for the community. This project is ongoing and, through collaborative efforts, keeps nursing career awareness in the forefront. PMID- 8710003 TI - Nurses and settlement houses: blueprint for neighborhood clinics. PMID- 8710004 TI - Use of a Delphi technique to prioritize clinical nursing research needs. AB - School of nursing faculty and nursing service personnel at an academic medical center in the South, collaboratively using a Delphi method, were able to prioritize the top three unit based clinical nursing research needs. The benefits were evident as research conducted at the staff level linked the research efforts with the quality improvement efforts on the Labor, Delivery and OB Receiving Units. The Delphi method, a consensus building technique, offered a clear, concise, accurate and non-threatening approach for staff nurses to participate in research. PMID- 8710005 TI - [Suxamethonium risky in intensive care]. PMID- 8710006 TI - [Risk factor assessment and prevention of psychosis]. PMID- 8710007 TI - [Long-term complications in diabetes mellitus: role of irreversible glycosylation products]. PMID- 8710008 TI - [Published effects of screening in parental and child health care]. AB - OBJECTIVE: A summary of research data on the evaluation of eight screening activities in Child Health Care in the Netherlands. DESIGN: Literature review. SETTING: Department of Public Health, Erasmus University Rotterdam, the Netherlands. METHOD: The literature was reviewed regarding screening effects on congenital hip pathology, congenital heart disease, retained testis, hydrocephalus, hearing loss, psychomotor retardation, visual disabilities, and phenylketonuria (PKU) and congenital hypothyroidism (CHT). RESULTS: For 8 types of screenings 17 targets could be established. Targets were defined in terms of a decrease of the adverse outcome of the diseases as a result of the screening. The efficacy of the screening for 3 of these targets (decrease of the adverse outcomes of PKU and CHT and needless orchidopexy) was convincingly established. For the other targets the available data were insufficient. Data on undesirable effects of screening were scarce. For 6 of the 8 screening activities some data on test properties were available. CONCLUSION: Research data on the effects of screening in Child Health Care in the Netherlands are incomplete. It is important to improve the scientific basis of these prevention activities. PMID- 8710009 TI - [The use of blood and blood products in total hip arthroplasty in 31 European hospitals, 1990/'91]. AB - OBJECTIVE: To compare the use of blood products and artificial colloids during total hip arthroplasty in European hospitals. DESIGN: Descriptive. SETTING: Europe. METHODS: During the period October 1990-October 1991 transfusion data were obtained about patients who underwent a planned total hip replacement for the first time. The investigations were performed in 31 teaching hospitals in ten countries of the EC, as a part of the ?Safe and good use of blood in surgery' (SANGUIS)-project. RESULTS: Red cells were ordered preoperatively in 97.4% of the 1647 cases and transfused in 81% (ranges among hospitals: 29-100). Hospitals in the Mediterranean area used more auto-transfusion than those in Central and Northern Europe. Plasma was transfused in 6% of the patients, predominantly in hospitals in southern European countries. Albumin was used especially in Central and Northern European countries. The reasons for red cell transfusion were stated in the medical records in 23% of the cases, for plasma transfusions in 7% and for albumin in 1%. Averaged transfusion-related costs were 192 ecu per patient (ranges per hospital: 60-383 ecu). CONCLUSION: Differences between European hospitals in the use of blood products for total hip arthroplasty are considerable. PMID- 8710010 TI - [Aging as a cause of hip fractures; a theoretic mathematical model]. AB - OBJECTIVE: To estimate the numbers of hip fractures among Dutch women caused by senescence in contrast to decreasing postmenopausal bone loss. DESIGN: A Markov chain model in a multi-state life table. SETTING: Department of Public Health, Erasmus University of Rotterdam, the Netherlands. METHOD: Cross-sectional data of prevalences of femoral neck bone mineral density, measured in an epidemiological survey (1993), were related to numbers of hospital admissions for hip fractures among women aged 55 and older, using a (published) relative risk for hip fracture of 2.6 (95% confidence limits: 1.9-3.6) per decrease by I SD of bone mineral density. We varied the prevalences of bone mineral density in the multi state life table and looked at changes in the expected number of fractures. RESULTS: If bone loss were to cease after the age of 55, this would decrease the number of fractures by 39.4% (30.4%-45.2% in sensitivity analysis). If bone loss were delayed by 7.5 years, the number of hip fractures would decrease by 14.8% (10.0% 19.9% after 5 to 10 years' delay). If compliance decreases by age (3% per year), then the number of hip fractures would decrease by 5.6% (3.4%-11.0% after 1%-5% decrease of compliance per year). CONCLUSION: Bone loss is only one of the many age related causes of hip fractures. Population based interventions targeting only bone mineral density, while ignoring comorbidity, have only a limited potential for preventing hip fractures. PMID- 8710011 TI - [Fatal arrhythmia following administration of suxamethonium in a postpartum woman subsequently found to have dystrophia myotonica]. AB - A 28-year-old woman died of irreversible ventricular fibrillation after administration of succinylcholine because of post partum curettage. Afterwards it became known she had undiagnosed myotonic dystrophy. She also had metabolic acidosis because of haemorrhagic hypovolaemia resulting in a probably elevated serum K+ concentration preoperatively. Succinylcholine given to patients with myotonic dystrophy may lead to life threatening hyperkalaemia and cardiac arrest. Therapy should be aimed at immediate lowering of serum K+ concentration with calcium, sodium bicarbonate and hyperventilation. PMID- 8710012 TI - [Patient-oriented education in family practice within medical education; curriculum of the Rijksuniversiteit Limburg]. PMID- 8710013 TI - [Controversial contraception: immunization against human chorionic gonadotropin]. PMID- 8710014 TI - [Controversial contraception: immunization against human chorionic gonadotropin]. PMID- 8710015 TI - [Synopsis of the standard 'Low backache' from the Nederlands Huisartsen Genootschap]. PMID- 8710016 TI - [Favorable effects of menopausal hormone substitution therapy on risk factors for ischemic heart disease]. PMID- 8710017 TI - [Primary hyperaldosteronism as a cause of hypertension; diagnosis]. PMID- 8710018 TI - [Dog bites: publications on risk factors, infections, antibiotics and primary wound closure]. PMID- 8710019 TI - [Atypical last menstruation: extrauterine pregnancy in the differential diagnosis]. PMID- 8710020 TI - [Lung transplantation]. PMID- 8710021 TI - [The introduction of lung transplantation in The Netherlands. Lung Transplantation Group Groningen]. PMID- 8710022 TI - [Hereditary mutations in the p53 tumor suppressor gene; significance for clinical practice. National Work Group Hereditary Mamma Carcinoma]. PMID- 8710023 TI - [Predictive testing for p53-mutations; ethical considerations. National Work Group Hereditary Mamma Carcinoma]. PMID- 8710024 TI - [The use of 3-dimensional models in craniofacial surgery]. AB - With the aid of axial CT scanning a polyurethane or resin copy of bony tissue of the craniofacial region of interest can be manufactured. 3-dimensional models are used in planning of osteotomies of the midface and forehead, where complex bone transpositions are to be performed, and in manufacturing complex implants. Different preoperative steps in several patients are demonstrated and the postoperative results are shown. Patient A was a 9-year-old girl who had radiotherapy as a baby for retinoblastoma which destroyed a large part of her face. In patient B, a 46-year-old woman, the whole zygomatic bone and three quarters of the floor of the orbit were among other things resected for a low grade osteosarcoma. Patient C had Goldenhar's syndrome (oculoauriculovertebral dysplasia). In all 3 patients reconstructive surgery was prepared with 3 dimensional cut models. PMID- 8710025 TI - [Diagnosis and treatment of extrauterine pregnancy in The Netherlands]. AB - OBJECTIVE: To inventory current policy in the diagnosis and treatment of ectopic pregnancy (EP) in the Netherlands. DESIGN: Questionnaire. SETTING: Academic Medical Centre, Amsterdam, the Netherlands. METHODS: A questionnaire was sent to all I30 departments of gynaecology in the Netherlands. RESULTS: The response rate was 94% (n = 122). Transvaginal ultrasonography was very widely used, whereas results of serum hCG measurements were only available within 24 h in 75% of the departments. Eighty-three per cent of the respondents screen for EP in high-risk patients. Laparoscopic surgery was used in 62% of the departments, open surgery in 9%, whereas in 28% the surgical technique depended upon the skills of the operating gynaecologist. If desire for pregnancy was present, 53% of the departments operated always conservatively, 2% always radically, whereas in 43% conservative surgery was performed only in case of a healthy ipsilateral tube. Methotrexate was used in 38% of the departments for persistent trophoblast, in 16% as primary treatment of interstitial pregnancies and in 13% as primary treatment for tubal pregnancy. CONCLUSION: Diagnostic strategies for EP in the Netherlands are homogeneous and in conformity with the state of the art in the literature. For treatment, laparoscopic techniques are widely used. In almost all departments, conservative surgery is the treatment of choice. Use of methotrexate is so far limited to treatment of persistent trophoblast after a failure of conservative surgery. PMID- 8710026 TI - [Results of 75 lung transplantations performed in Groningen in 1990-1995. Lung Transplantation Group Groningen]. AB - OBJECTIVE: To describe the development of the lung transplantation programme in Groningen, and the results of single and bilateral lung transplantations in the first 75 consecutive patients, up to December 1995. DESIGN: Retrospective. SETTING: Academic Hospital Groningen, the Netherlands. METHODS: The results of the lung transplantation programme were evaluated retrospectively. RESULTS: In November 1990 the first unilateral lung transplantation was performed in Groningen in a patient with pulmonary fibrosis. In February 1991 a national lung transplantation programme for the Netherlands was instituted in Groningen by the government. Of 500 patients referred from all over the Netherlands from 1990 to December 1995, 75 were transplanted, 16 unilaterally and 59 bilaterally. The actuarial survival for all patients was 85% after 1 year and 72% after 2 years. After transplantation 16 patients died (21%) after 15 months follow-up (median). Early mortality (5%) was caused by graft failure, late mortality (16%) by chronic rejection and lymphoproliferative disease. The mean time on the transplantation waiting list was 9.3 months; it increased during the programme. The limiting factor for further expansion of the programme was caused by donor scarcity. The lungs from only 16% of the multiorgan donors reported by Eurotransplant to our centre could be transplanted. CONCLUSION: The results of the lung transplantation programme in Groningen are good but with an increasing number of lung transplantation centres in the Eurotransplant region the further development of lung transplantation in the Netherlands will depend mainly on the availability of lung donors from the Netherlands. PMID- 8710027 TI - [Herpetic esophagitis in 5 immunocompetent patients]. AB - In five immunocompetent patients, 3 of whom were moderately ill, herpes simplex virus (HSV) oesophagitis was diagnosed. HSV oesophagitis is a frequent infection in immunocompromised hosts. Nevertheless the English medical literature (Medline) contains at least 33 cases of HSV oesophagitis in immunocompetent persons over the period 1983 to 1993. Odynophagia, dysphagia and chest pain are common symptoms. Endoscopy is necessary for diagnosis: brush cytology, biopsy for histology and viral culture are the tools for identification of herpes simplex as the cause of the oesophagitis. HSV oesophagitis in an immunocompetent patient is an acute but self-limiting disease. Nevertheless acyclovir per os or intravenously may be recommended if started early after onset of symptoms. PMID- 8710028 TI - [Advising lawyers; a medicolegal lesson]. PMID- 8710029 TI - [Xanthelasma palpebrarum]. PMID- 8710030 TI - [Prevalence of hemorrhages caused by vitamin K deficiency in The Netherlands, 1992-1994]. PMID- 8710031 TI - Abstracts of the Internistendagen Veldhoven, Netherlands, 25-26 April 1996. PMID- 8710032 TI - "Spontaneous" hypoglycaemia: facing a hydra. PMID- 8710033 TI - Similar fragmentation and dissolution after extracorporeal shock wave lithotripsy in eutrophic and overweight patients with gallstones. AB - The aim of this study was to assess the relative importance of overweight and adjuvant treatment with bile acids to obtain fragment dissolution and clearance after biliary extracorporeal shock wave lithotripsy (BESWL), in eutrophic and overweight patients with gallbladder stones. During a 3-year period 103 patients were treated with BESWL. Patients were strictly selected in terms of the number and type of stones. Of the total of 103 patients, 53 were women and 50 men. Mean age was 50.8 +/- 14.3 (range 17 to 86) years. Body mass index was calculated for each patient. All patients underwent BESWL with an electromagnetic device. Medical therapy for stone dissolution included the combination of chenodeoxycholic acid plus ursodeoxycholic acid (50% + 50%), at a total dose of 10-15 mg/day. Of the 103 patients, 45 were eutrophic and 58 were considered overweight. Age was similar in both groups. Number of lithotripsy sessions in eutrophic patients was 1.96 +/- 1.07 and 1.88 +/- 0.84 in overweight patients, but no statistical differences were found. Complete clearance of calculi was observed in more than 50% of the cases after 6 months, and reached 98% after 22 months of therapy in both groups. Total clearance of calculi was similar in patients with single and multiple stones. In conclusion, our results suggest that the time required to obtain complete gallstone clearance after BESWL is similar in eutrophic and overweight patients with gallbladder stones, and also that adjuvant therapy with bile acids plays an important role in gallstone fragment dissolution in these patients. PMID- 8710034 TI - Non-islet-cell tumour induced hypoglycaemia: a case report and review of literature. AB - A 54-year-old man presented with a poorly differentiated adenocarcinoma of the rectum with multiple metastases to the liver. During hospitalization the patient developed periods of hypoglycaemia due to production of "big" IGF-II by the tumour. Possible pathophysiological mechanisms of non-islet-cell tumour-induced hypoglycaemia are discussed. PMID- 8710035 TI - Spontaneous hypoglycaemia in chronic renal failure. AB - Spontaneous hypoglycaemia in renal failure occurs more frequently than is considered generally. The pathogenesis is complex. Understanding the underlying mechanisms is of interest to all practitioners attending patients with renal failure. A case report is presented describing a patient with 'spontaneous' hypoglycaemia, without any illness other than renal failure. The various mechanisms contributing to or causing hypoglycaemia are reviewed. PMID- 8710036 TI - Reversal of fulminant-hepatitis-associated hypoglycaemia at the anhepatic stage during liver transplantation. AB - Hypoglycaemia is a known complication of fulminant hepatic failure. Massive destruction of liver tissue, along with hyperinsulinism and defective glucose storage in extrahepatic organs are some of the mechanisms contributing to the hypoglycaemia. We describe here a case of reversal of fulminant-hepatitis associated hypoglycaemia at the anhepatic stage of liver transplantation. It is suggested that non-insulin hypoglycaemic factors secreted by the damaged liver may be responsible for this complication. PMID- 8710037 TI - Recurrent hypoglycaemia caused by the insulin autoimmune syndrome: the first Dutch case. AB - The insulin autoimmune syndrome is extremely rare outside of Japan. We describe a 45-year-old Caucasian woman who presented with fasting hypoglycaemia and weight gain. The presence of fasting insulin concentrations in excess of 1000 mU/l, very low C-peptide concentrations during hypoglycaemic attacks and high titres of insulin autoantibodies led to the diagnosis. Treatments aimed at decreasing endogenous insulin secretion by either dietary intervention alone or in combination with acarbose, octreotide or diazoxide had only limited success, while a 2-week course of immunosuppression with prednisone was without any antihypoglycaemic effect. PMID- 8710038 TI - Hydralazine-induced constrictive pericarditis. AB - A 59-year-old man was diagnosed as having constrictive pericarditis 17 months after a typical hydralazine-induced autoimmune syndrome. This late complication of hydralazine has been reported only once. Ten years later the patient was found to have anti-neutrophil cytoplasmic antibodies directed against myeloperoxidase. PMID- 8710039 TI - The bleeding risk in chronic haemodialysis: preventive strategies in high-risk patients. AB - Haemodialysis (HD) patients are at an increased risk of bleeding because of uraemic bleeding tendency and systemic anticoagulation caused by intermittent heparinization. Additional risk factors may be aspirin or coumarin use for the prevention of fistula thrombosis, diffuse intravascular coagulation, recent trauma, postsurgical state, inadequate control of hypertension, gastrointestinal lesions, diabetic retinopathy, renal cystic disease, and uraemic pericarditis. In HD patients with an active bleeding focus blood transfusion, desmopressin acetate (DDAVP), conjugated oestrogens, and dialysis treatment can limit the bleeding risk. Strategies to reduce the bleeding risk conveyed by systemic anticoagulation during HD are regional heparin anticoagulation and minimal heparinization. In our opinion, dialytic modalities that completely preclude systemic anticoagulation, i.e. peritoneal dialysis (PD), heparin-free HD, and HD with regional anticoagulation with prostacyclin, mesilates, or citrate, are indicated for patients who are actively bleeding, or who are within 3 days from a bleeding episode, or a surgical or accidental wound, or who are less than 2 weeks from cerebral or retinal haemorrhage, and for patients with incompletely treated proliferative diabetic retinopathy or uraemic pericarditis. PMID- 8710040 TI - Uveitis during treatment of disseminated Mycobacterium avium-intracellulare complex infection with the combination of rifabutin, clarithromycin and ethambutol. AB - Three patients with a cellular immunodeficiency were treated with rifabutin, clarithromycin and ethambutol for a disseminated infection with Mycobacterium avium-intracellulare complex (MAC). The patients developed uveitis, sometimes in combination with a transient rash, arthralgia, arthritis, jaundice and pseudojaundice. It seems likely that these reactions were caused by rifabutin, alone or together with other drugs such as clarithromycin. These adverse reactions probably depend on the dose, metabolism and excretion of the drug. Inhibition of cytochrome P450 seems to be an important mechanism. PMID- 8710041 TI - Partial unilateral phrenic nerve paralysis caused by a large intrathoracic goitre. AB - Intrathoracic goitres may cause a variety of symptoms caused by compression of the trachea, neural structures, blood vessels and the oesophagus. A case history is presented of a patient with a recurrent goitre after subtotal thyroidectomy who displayed partial unilateral phrenic paralysis, which subsided after a second subtotal thyroidectomy. Compression of the phrenic nerve appears to be a very rare manifestation of an intrathoracic goitre and thus far has never been reported. PMID- 8710042 TI - Serum LDH: a marker of disease activity and its response to therapy in idiopathic pulmonary fibrosis. AB - In the following case report the determination of serum lactate dehydrogenase (LDH)-a simple though non-specific test-reflects changes of disease activity and clinical improvement during treatment with cyclophosphamide and a tapered dose of prednisone from 100 to 0 mg daily in a 43-year-old woman with idiopathic pulmonary fibrosis. The trend observed in this case indicates that the serum LDH activity may be directly proportional to the extent of this diffuse inflammatory pulmonary disease. PMID- 8710043 TI - Unexplained high activity of aspartate aminotransferase (EC 2.6.1.1): a case report. AB - We report a case of unexplained and persistently elevated ASAT activity in serum. This appeared to be due to a macro-enzyme of ASAT, as was demonstrated by protein A-Sepharose treatment of the serum sample. Although macro-ASAT seems to be clinically irrelevant, unexplained raised ASAT activity without acknowledgement of its macroenzyme nature may lead to unnecessary diagnostic procedures. PMID- 8710045 TI - Suggestive evidence for bromocriptine-induced pleurisy. AB - Pleurisy of initially unknown origin was found in a patient who was treated with bromocriptine for Parkinson's disease for 6 years. At presentation, bilateral pleural thickening existed that caused severe restriction of pulmonary function. There were an elevated erythrocyte sedimentation rate, polyclonal hypergammaglobulinaemia, increased levels of acute phase proteins and anaemia. After withdrawal of the bromocriptine the patient's complaints as well as the laboratory parameters markedly improved. Further loss of pulmonary function did not occur. However, the pleural thickening did not resolve, not even upon subsequent corticosteroid treatment, probably due to fibrosis. Together, these findings strongly suggest a causative role of bromocriptine. The results of the laboratory studies suggested an immunopathogenetic mechanism, but in vitro lymphocyte-proliferation studies and skin patch tests with bromocriptine were negative. Bromocriptine should be considered as a cause of pleurisy. The drug must be stopped immediately upon the occurrence of pleural thickening in order to prevent impairment of pulmonary function. In addition, periodic laboratory and X ray studies in patients on long-term bromocriptine treatment should be considered. PMID- 8710044 TI - Vertebral bone destruction in sickle cell disease: infection, infarction or both. AB - Infectious and vaso-occlusive vertebral bone and joint destruction in two patients with sickle cell disease (SCD) are featured by H-shaped vertebrae, kyphotic angulation, osteolysis of endplates and collapse of intervertebral discs as shown by X-ray films and magnetic resonance imaging. Staphylococcal serology supported the diagnosis of staphylococcal osteomyelitis/spondylo-discitis in both SCD patients. The difficulties of establishing the causes and treatment of the osteoarthropathy in these particular cases are discussed in the light of the literature. PMID- 8710046 TI - Cytochrome P450 isoenzymes--importance for the internist. AB - The studies with CYP isoenzymes give hope that in the near future it will be possible to predict the clinical pharmacokinetics of drugs, if their oxidation can be assigned to the activity of one or more well-characterized CYP isoenzymes (or if their metabolism is catalyzed by other well-studied enzymes). This strategy will reduce animal experimentation. Interindividual variability in drug metabolism translates into variability in drug efficacy and toxicity. Establishing the status of drug-metabolizing enzymes will therefore assist in making predictions of pharmacologic and toxicologic responses to drugs. Many clinically relevant drug-drug interactions can now be readily rationalized in terms of the substrate and inhibitor specificities of individual human CYP isoenzymes. The next 5 years should reveal whether selective inhibitors of xenobiotic-metabolizing CYP's can be used therapeutically in the treatment or prevention of cancer and various endocrine disorders, in analogy to the use of influencing steroidogenic CYP's. PMID- 8710047 TI - The spectrum of liver disease in systemic lupus erythematosus. AB - Patients with systemic lupus erythematosus (SLE) have a 25-50% chance of developing abnormal liver tests in their lifetime. This percentage does not include unconjugated hyperbilirubinaemia due to haemolysis associated with SLE, or elevated aspartate-aminotransferase caused by SLE-associated myositis. The most common cause is drug-induced hepatitis, while mild, predominantly lobular but sometimes also portal and periportal-hepatitis reflecting SLE activity is another possibility. Other liver disease in SLE can be related to thrombotic events, whether or not associated with the lupus anticoagulant, including Budd Chiari syndrome and veno-occlusive disease. Other liver abnormalities have been more or less frequently associated with SLE, such as nodular regenerative hyperplasia, perihepatitis, and hepatic or splenic rupture. Also viral hepatitis, obstructive jaundice, autoimmune hepatitis, primary biliary cirrhosis, granulomatous hepatitis, cryptococcus infection of the liver, chronic hepatitis with IgA or IgD deficiency, porphyria or idiopathic portal hypertension co existing with SLE have been described. PMID- 8710048 TI - [Chronic renal insufficiency 2000. Livre Blanc de la Nephrologie]. PMID- 8710049 TI - Production and characterization of a monoclonal antibody against medulloblastoma derived cell line. AB - A murine monoclonal antibody of the immunoglobulin M subclass, termed M1C, was prepared by fusion of P3X63Ag8U.1 mouse myeloma cells with spleen cells from BALB/c mice immunized with the recently established human medulloblastoma cell line MED-3. MED-3 cells are unique in expressing both neuronal and glial phenotypes. The antigen recognized by M1C was expressed predominantly in the cytoplasm of antigen-bearing cells, but was also present in lower concentration on the cell surface. Flow cytometry showed M1C binding to MED-3 cells increased during the G2/M phase, and decreased after induction of differentiation with N6, O2-dibutyryl cyclic adenosine monophosphate. Biochemical analysis indicated that M1C recognized a 25-kd protein. Immunoperoxidase staining of cultured cell lines showed that although M1C recognized a large panel of malignant tumor-derived cell lines, the staining of malignant glioma-derived cell lines was strongest. Immunohistochemical examination of various human tissues revealed that M1C reacted strongly with malignant gliomas but did not recognize low grade astrocytomas, neuroblastomas, or tumors from other organs. Cross-reactivity with normal tissues was restricted to peripheral blood lymphocytes. The immunoreactivity of M1C was quite different from the two previously described monoclonal antibodies against medulloblastoma. M1C is the first reported MoAb against medulloblastoma, which recognizes a differentiation- and proliferation dependent antigen expressed in malignant gliomas in particular, and may be useful in determining the differentiation and proliferation features of medulloblastomas and gliomas. PMID- 8710050 TI - Growth and immunogenicity of human glioma in severe combined immunodeficiency human mice. AB - The severe combined immunodeficiency (SCID) mouse was investigated as a model system to study the growth and immunogenicity of human gliomas. Human glioma cell lines U-251MG, KNS-42, SF-188, A-172, and T-98G were injected subcutaneously into SCID mice. The cell lines U-251MG and KNS-42 grew as large, subcutaneous masses; SF-188, A-172, and T-98G did not grow. Glioma-immune system interactions were studied by the transplantation of human peripheral blood lymphocytes into tumor bearing SCID mice. In the resultant SCID-human (SCID-hu) mice, transplanted lymphocytes infiltrated into the subcutaneously growing tumors and expressed the surface markers of human B, T, and natural killer cells. The SCID-hu mouse is a potentially powerful model to study the basic tumor biology of some human gliomas and also represents a useful screening system to evaluate experimental immunotherapies for brain tumors. PMID- 8710051 TI - Effect of thyrotropin-releasing hormone on Na(+)-K(+)-Adenosine triphosphatase activity following experimental spinal cord trauma. AB - The effect of thyrotropin-releasing hormone (TRH) on spinal cord Na(+)-K(+ adenosine triphosphatase (Na+-K+-ATPase) activity after spinal cord injury was evaluated in rats. The rats were injured by compression of the cord at T-10 for 1 minute with a 50-g clip. Saline in the placebo group (n = 8) and TRH (0.6 mg per dose) in the TRH group (n = 9) were administered intraperitoneally as bolus injections in two doses, at 45 and 120 minutes after the injury. The Na(+)-K(+) ATPase activity level in the TRH group was significantly higher (p = 0.024) than in placebo group. These results indicate a possible role for TRH treatment in spinal cord injury. PMID- 8710052 TI - Coagulation and fibrinolysis study after local thrombolysis of a cerebral artery with urokinase. AB - Coagulation and fibrinolysis factors were studied in six patients after local thrombolysis with urokinase (720,000 IU). Transient abnormalities, such as prolonged prothrombin time, decreased plasminogen and alpha 2-antiplasmin activities, decreased fibrinogen, and increased fibrin degradation products were seen on the day after thrombolysis, but tended to return to the normal range on the 4th day except for one patient who suffered from disseminated intravascular coagulation. Antithrombin III activity did not change so much. Therefore, the dosage of urokinase should be as low as possible to prevent fluctuations in the coagulation and fibrinolysis system. PMID- 8710053 TI - Surgical treatment of children with medically intractable epilepsy--outcome of various surgical procedures. AB - Outcome after temporal lobe resection, extratemporal resection, and corpus callosotomy was studied in 33 children aged 15 years or less with medically intractable epilepsy. Seizure-free control was achieved in 67% (6/9) of pediatric patients versus 67% (60/90) of adult patients receiving temporal lobe resection and in 33% (4/12) of pediatric patients versus 25% (7/28) of adult patients receiving extratemporal resection. Among patients suffering from drop attacks and generalized convulsive seizures, 42% (5/12) of pediatric patients versus 25% (6/24) of adult patients receiving corpus callosotomy became seizure-free. The most prominent histological diagnoses in temporal lobe resections were three cases of mesial temporal sclerosis, four of ganglioglioma, and two of neuronal migration disorders. Neuronal migration disorders were the main histology in extratemporal resections. Four of five patients with behavioral problems treated by corpus callosotomy exhibited improved hyperactivity and/or attention deficits. There was no surgical morbidity or mortality. The outcome of pediatric patients was at least as favorable as that of adult patients, and improvement of behavioral disturbances could be expected after surgery. Surgical treatment is an effective and safe procedure for children with medically intractable epilepsy. PMID- 8710054 TI - Modified petrosal approach using cosmetic osteoplastic craniotomy with NEURO-SAT guidance. AB - The petrosal approach has become a routine procedure for petroclival lesions, but there are some disadvantages such as the time-consuming craniotomy during surgery and depressed deformity of the mastoid area after surgery. To solve these disadvantages, we modified the petrosal approach. Before surgery, the three dimensional points of the sigmoid sinus and semicircular canals are calculated on computed tomography scan and are input to a computer. A single temporooccipital bone flap is made, and the outer table of bone overlying the mastoid is preserved by forming a narrow groove with a small air drill and cutting the outer table with a chisel. After removing these two free bone flaps, mastoidectomy is performed guided by NEURO-SAT (neuronavigation by computer-assisted frameless stereotaxy). The mastoid bone can be drilled out safely and quickly. The two bone flaps are connected and replaced at the end of the procedure. Osteoplastic petrosal craniotomy guided by NEURO-SAT can achieve a quick craniotomy and satisfactory cosmetic result. PMID- 8710055 TI - Posterior inferior cerebellar artery aneurysm located in the spinal canal--case report. AB - A 30-year-old male presented with subarachnoid hemorrhage, due to a rare peripheral posterior inferior cerebellar artery aneurysm located in the spinal canal. Angiography and magnetic resonance imaging disclosed the aneurysm of 15 mm diameter at the C-1 level. After aneurysmal neck clipping and ventriculoperitoneal shunt implacement, he was discharged without neurological deficit. Histological examination revealed no layer formation, only hemorrhage and organization in the aneurysmal wall. This case may be an example of giant aneurysm formation caused by repeated bleeding and organization. PMID- 8710056 TI - Central neurocytoma with lipofuscin granules--case report. AB - A 30-year-old male presented with a large calcified intraventricular mass. The tumor was completely resected via the transventricular route. Light microscopy showed the tumor was composed of small round cells with perinuclear halo, typical of oligodendroglioma. Electron microscopy revealed marked neuronal differentiation, including neurosecretory granules, well-formed numerous synapses, and lipofuscin granules. The tumor was diagnosed as well-differentiated central neurocytoma. PMID- 8710057 TI - Cervical ventral epithelial cyst treated by anterior corpectomy--case report. AB - A 15-year-old boy presented with a cervical ventral epithelial cyst manifesting as intractable cervical pain. Neurological examination revealed weakness of his left grasp and increased deep tendon reflexes. The anterior approach was selected because the lesion was located directly ventral to the spinal cord. He underwent C4-5 anterior corpectomy and subtotal resection of the cyst wall. He made a good recovery without additional deficits. Most previously reported cases have been treated using the posterior approach despite the ventral location of the lesion. The anterior approach offers a safer and more effective means of treating such lesions because it allows direct access to the lesion without manipulation of the spinal cord. PMID- 8710058 TI - Delivery of primary neurosurgical care in developing countries--scope for mutual cooperation. AB - As with other specialized services there is an ever increasing gap in neurosurgical care between developed and developing countries. The need for neurosurgery has always been present in developing countries, but the recent introduction of computed tomography has identified many previously undetected cases, increasing the number of patients seeking treatment. However, developing countries suffer severe shortages of trained manpower, proper equipment, and expertise to initiate the services to meet this demand. In contrast, some developed nations are experiencing a surplus of neurosurgeons leading to problems in case exposure. A well-designed cooperative project between these countries can answer some of these problems, becoming an ideal example of mutual benefit. PMID- 8710059 TI - Clinical research criteria for the diagnosis of progressive supranuclear palsy (Steele-Richardson-Olszewski syndrome): report of the NINDS-SPSP international workshop. AB - To improve the specificity and sensitivity of the clinical diagnosis of progressive supranuclear palsy (PSP, Steele-Richardson-Olszewski syndrome), the National Institute of Neurological Disorders and Stroke (NINDS) and the Society for PSP, Inc. (SPSP) sponsored an international workshop to develop an accurate and universally accepted set of criteria for this disorder. The NINDS-SPSP criteria, which were formulated from an extensive review of the literature, comparison with other previously published sets of criteria, and the consensus of experts, were validated on a clinical data set from autopsy-confirmed cases of PSP. The criteria specify three degrees of diagnostic certainty: possible PSP, probable PSP, and definite PSP. Possible PSP requires the presence of a gradually progressive disorder with onset at age 40 or later, either vertical supranuclear gaze palsy or both slowing of vertical saccades and prominent postural instability with falls in the first year of onset, as well as no evidence of other diseases that could explain these features. Probable PSP requires vertical supranuclear gaze palsy, prominent postural instability, and falls in the first year of onset, as well as the other features of possible PSP. Definite PSP requires a history of probable or possible PSP and histopathologic evidence of typical PSP. Criteria that support the diagnosis of PSP, and that exclude diseases often confused with PSP, are presented. The criteria for probable PSP are highly specific, making them suitable for therapeutic, analytic epidemiologic, and biologic studies, but not very sensitive. The criteria for possible PSP are substantially sensitive, making them suitable for descriptive epidemiologic studies, but less specific. An appendix provides guidelines for diagnosing and monitoring clinical disability in PSP. PMID- 8710060 TI - Ten steps in characterizing and diagnosing patients with peripheral neuropathy. AB - Cost-effective approaches for arriving at correct diagnoses should be used. Herein, we compare three approaches for the differential diagnosis of a common neurologic syndrome, peripheral neuropathy. In the "shotgun" approach, a standard battery of hematologic, biochemical, serologic, antibody, enzyme, molecular genetic, and other tests are ordered after the presence of a neuropathy is established, without a detailed characterization of the neuropathy. In the "gestalt" approach, the specific variety of neuropathy is identified by recognition of a clinical pattern of symptoms, course, or disease associations. The "10-step" approach, encompassing components of the two other methods and adding others, evolved from our assessment of many patients with peripheral neuropathy. In the 10-step approach, the history and neurologic examination place the patient's disorder into one of perhaps 21 anatomic-pathologic patterns. Next, electrophysiologic and other tests confirm the correctness of this anatomic pathologic patterns. Finally, a series of evaluations exclude or include an increasingly shorter list of diagnoses until only one likely one remains or the disorder remains undiagnosed. We advocate the 10-step over the shotgun or gestalt approach, because it emphasizes careful initial characterization of neuropathy and emphasizes use of a logical step-by-step inclusion or exclusion to arrive at a short list of diagnostic possibilities. The approach depends ultimately on the judgement of adequately trained and experienced physicians, not on the results of single tests. The 10-step approach is not a mindless algorithm, leading inevitably to the correct diagnosis, but depends on judgment based on extensive background and neurobiologic and clinical knowledge and training. PMID- 8710061 TI - Eyeball pressure testing in the evaluation of serious bradyarrhythmias in Guillain-Barre syndrome. AB - OBJECTIVE: To investigate the usefulness of eyeball pressure testing (EP) as an indicator for impending serious bradyarrhythmias in patients with Guillain-Barre syndrome (GBS) and its relationship to motor disability. BACKGROUND: Autonomic dysfunction is a common complication in GBS and accounts for a significant number of deaths. Serious bradyarrhythmias are thought to occur only in severe cases but are difficult to predict. METHODS/DESIGN: In 13 consecutive patients with GBS aged 29 to 70 years, 156 EP (6 to 19 per patient) were done serially for up to 1 year. Bilateral moderate pressure was manually applied and sustained for 25 seconds or until abnormal bradycardia developed, defined as heart rate below 40 beats per minute. Disability was graded by a score from 0 to 5 (DS). RESULTS: Four of 13 patients (DS 2/2/3-4/5) showed abnormal sensitivity to EP at least once. In two of them, vagal overreactivity could be demonstrated repeatedly, which gradually resolved within 4 and 10 days. In one patient with a rapid progressive course requiring early cardiopulmonary resuscitation, a highly abnormal EP could be recorded until 1 day after heart arrest. Another patient (DS 3-4) with abnormal EP required cardiac pacing twice because of significant bradycardia. The only other event necessitating pacing occurred in a severely disabled patient (DS 5-4) who never showed abnormal EP. CONCLUSIONS: Vagal overreactivity could be demonstrated in approximately 30% of our patients. It was not restricted to severe motor impairment and was also present in mild-to moderately disabled patients. In this regard, EP may be a simple and useful bedside test to indicate an increased risk of developing serious bradyarrhythmias in patients with GBS. PMID- 8710062 TI - Gait of a deafferented subject without large myelinated sensory fibers below the neck. AB - We evaluated the gait pattern of a deafferented subject who suffered a permanent loss of large sensory myelinated fibers below the neck following an acute episode of purely sensory neuropathy 21 years ago. The subject has developed several strategies to achieve a secure gait, namely: (1) a reduction of the degrees of freedom by freezing the knee articulations during the stance phase, (2) a preservation of body balance by enlarging his base of support, and (3) visual monitoring of his step by stabilizing the head-trunk linkage together with a characteristic forward tilt. As a result, the gait of the deafferented subject lacks the fluidity of normal gait. Compared with normal subjects, the gait pattern of the deafferented subjects is characterized by a shorter cycle length, a longer cycle duration, a slower speed, and a lower cadence. Using a dual-task paradigm, the attentional demands for walking were particularly important (as indexed by longer probe reaction times) during the double-support phase, suggesting that the deafferented subject uses the double-support phase as a transitory stable phase to update cognitively the postural features necessary for generating his next step. PMID- 8710064 TI - Sustained myoglobinuria: the presenting manifestation of dermatomyositis. AB - We describe two patients with sustained myoglobinuria that did not resolve with supportive treatment. After we established a diagnosis of dermatomyositis and started treatment with corticosteroids, there was rapid resolution of the myoglobinuria. Well-documented cases of inflammatory myopathy causing myoglobinuria are rare, but their prompt recognition has important therapeutic implications. PMID- 8710063 TI - Motor neuropathy due to docetaxel and paclitaxel. AB - Paclitaxel and docetaxel are novel chemotherapeutic agents that promote the polymerization and inhibit the depolymerization of microtubules. Sensory neuropathy is common with these agents, particularly paclitaxel. We evaluated 64 patients treated with these drugs; 54 were followed prospectively. Eleven (17%, including six of the 54 prospectively followed patients) developed muscle weakness that was predominantly proximal. The weakness was idiosyncratic, occurring at any stage of treatment, had a variable course, and was reversible upon cessation of drug. All patients developed symptoms or signs of taxane induced sensory neuropathy. Weakness was likely neuropathic in origin; electrodiagnostic studies suggested a distal axonopathy in some patients and proximal denervation (anterior horn cell or nerve root) in other. PMID- 8710065 TI - Interferon alpha-2a treatment of relapsing-remitting multiple sclerosis: disease activity resumes after stopping treatment. AB - We evaluated the long-lasting effects of systemic high-dose recombinant interferon alpha-2a (rIFNA) in relapsing-remitting (RR) MS after discontinuing treatment in a single-blind randomized placebo-controlled trial with 20 RR clinically definite MS patients using either nine million IU intramuscular rIFNA (n = 12) or placebo (n = 8) every other day for 6 months. Follow-up continued for a further 6 months without IFN treatment. In rIFNA-treated patients, main outcome measures, significantly different from placebo during treatment, returned, after discontinuing treatment, to values similar to placebo or baseline. Active MRI lesions per patient increased from 0.08 +/- 0.08 to 1.2 +/- 0.4 (p < 0.02), number of patients with clinical MRI signs of disease activity from 2 of 12 to 8 to 12 (P < 0.04), lymphocyte IFN gamma production from 3.0 +/- 0.7 to 12.4 +/- 2.2 IU/mL (p < 0.01), lymphocyte tumor necrosis factor alpha production from 5.8 +/- 0.9 to 18.9 +/- 6.3 pg/mL (p < 0.05). All side effects of rIFNA treatment disappeared after discontinuing the drug. The reduction of clinical MRI signs of disease activity and the immunologic effects were temporary and restricted to the period of rIFNA administration. The depression of many immunologic and clinical MRI responses during drug administration and their simultaneous return to baseline after discontinuing the drug strongly argue all observed changes were related to drug administration. PMID- 8710066 TI - Validation of the functional assessment of multiple sclerosis quality of life instrument. AB - Based on scientific literature and interviews with clinicians and patients, we developed a quality of life instrument for use with people with MS called the Functional Assessment of Multiple Sclerosis (FAMS). The initial item pool consisted of 88 questions: 28 from the general version of the Functional Assessment of Cancer Therapy quality of life instrument, plus 60 generated by patients, providers, and literature review. The validation samples comprised a mail survey cohort (N = 377) and a clinical cohort (N = 56). Both cohorts provides evidence for internal consistency of the derived subscales, test-retest reliability, content validity, concurrent validity, and construct validity. Principal components and Rasch measurement model analyses were applied sequentially to survey sample data, reducing test length to 44 questions, divided into six subscales: mobility, symptoms, emotional well-being (depression), general contentment, thinking/fatigue, and family/social well-being. Fifteen initially rejected questions were added back as miscellaneous (unscored) questions for their potential clinical and empirical value. The mobility subscale was strongly predictive of the Kurtzke Extended Disability Status Scale and the Scripps Neurologic Rating Scales. The other five subscales were not, indicating they measure aspects of patient quality of life not captured by the neurologic exam. The final 59-item English language instrument (FAMS version 2) is available for inclusion in clinical trials and clinical practice. PMID- 8710067 TI - The effect of spectrum bias on the utility of magnetic resonance imaging and evoked potentials in the diagnosis of suspected multiple sclerosis. AB - BACKGROUND: Spectrum bias is the tendency for the effectiveness of a test (or treatment) to vary as a function of clinical, pathologic, or comorbid variables including disease severity. Our null hypothesis was that within the population of suspected multiple sclerosis (MS) patients, there would be no difference in MRI and evoked potential (EP) sensitivity and specificity between those with mild MS versus those with more severe clinical signs of the disease, i.e., that spectrum bias is not a factor. METHODS: Three hundred three patients with suspected MS were evaluated by a board-certified neurologist, then scanned with MRI. Two hundred four patients also received EP testing. RESULTS: The sensitivity of MRI in patients with suspected MS was 58 percent with a false-positive rate of 9%. The overall sensitivity was 64% in the probable and 45% in the possible group. In the low pretest probability group sensitivity was 20%, and it was 70% in the high pretest probability group. These differences in sensitivity are statistically significant (p < 0.03). In contrast, the specificity between groups did not differ significantly. EP sensitivity was 69% in the high probability subgroup and 5% in the low probability subgroup. (p < 0.01). CONCLUSION: In this study, both EP and MRI results demonstrated an association between disease frequency, disease severity, and test sensitivity with greater disease frequency and intensity suggesting more impressive diagnostic test performance. The distorting effect of the variable clinical severity on MRI and EP sensitivity in suspected MS underscores that diagnostic tests perform differently in different groups of patients. PMID- 8710068 TI - Subacute leukoencephalitis caused by CNS infection with human herpesvirus-6 manifesting as acute multiple sclerosis. AB - Several recent reports have documented the neuroinvasiveness of human herpesvirus 6 (HHV-6) in infants with primary HHV-6 infections, in children and adults with AIDS, in recipients of bone marrow transplants, and in immunologically intact adults and children. CNS infections with HHV-6 can be subacute and are frequently associated with diffuse or multifocal demyelination. We analyzed the CNS tissues of a young woman who died of a demyelinative disease, which was clinically and histopathologically diagnosed as acute multiple sclerosis, for active HHV-6 infection by immunohistochemical staining. The tissues contained a dense and disseminated active HHV-6 infection that was intimately related to the pathologic changes present. PMID- 8710069 TI - Follow-up study of risk factors in progressive supranuclear palsy. AB - The cause of progressive supranuclear palsy (PSP) is not known and has been little studied. The one previous controlled epidemiologic survey, performed at our center in 1986, found small-town experience and greater educational attainment as PSP risks, but, in retrospect, these results may have been produced by ascertainment bias. Since that time, several anecdotal reports have implicated heredity and various environmental exposures in the cause of some cases of PSP. To clarify the results of the previous study and to evaluate the more recently implicated candidate factors in a controlled fashion, we mailed a validated 69 item questionnaire to 91 personally examined patients with PSP and 104 unmatched controls with other neurologic conditions for which they had been referred to our tertiary neurologic center. We were able to match 75 subjects from each group by year of birth, sex, and race and subjected them to a separate matched-pair analysis. We allowed surrogates to supply any or all of the responses. Questions concerned hydrocarbon, pesticide, and herbicide exposure; urban/rural living; auto repair and other occupations; head trauma; educational attainment; maternal age; and family history of PSP, parkinsonism, dementia, and other neurologic conditions. A statistically significant finding was that patients with PSP were less likely to have completed at least 12 years of school (matched odds ratio = 0.35, 95% CI = 0.12-0.95, p = 0.022; unmatched odds ratio = 0.44, 95% CI = 0.21 0.89, p = 0.020). We hypothesize that this result may be a proxy for poor early life nutrition or for occupational or residential exposure to an as-yet unsuspected toxin. Future studies should examine these potential risk factors in PSP. PMID- 8710070 TI - Risk of Parkinson's disease among first-degree relatives: A community-based study. AB - OBJECTIVE: To determine the relative risk (RR) and cumulative incidence of idiopathic Parkinson's disease (PD) in first-degree relatives of PD patients compared with relatives of controls from the same geographic region. DESIGN: A family history questionnaire was used to obtain information on all first-degree relatives of cases and controls. A subset of these first-degree relative was also examined. A Cox proportional hazards model with double-censoring techniques for missing information was used to model the RR for PD, adjusting for gender, ethnicity, and relationship to proband. RESULTS: A total of 1,458 first-degree relatives of 233 PD patients were 2.3 times as likely (95% CI = 1.3 to 4.0) as 7,834 relatives of 1,172 controls to develop PD. The cumulative incidence of PD to age 75 among first-degree relatives of PD patients was 2% compared with 1% among first-degree relatives of controls. The risk in male first-degree relatives was higher than in female relatives (RR = 2.0, 95% CI = 1.1 to 3.4) and the risk in relatives of Caucasians was higher than in African-Americans and Hispanics (RR = 2.4, 95% CI = 1.4 to 4.1). Risk for siblings and parents of probands was similar. CONCLUSIONS: Susceptibility to PD is increased in first-degree relatives of both sporadic and familial cases. The pattern of inheritance and the relationship between genetic and environmental risk factors warrant further study. PMID- 8710071 TI - Clinical analysis of 17 patients in 12 Japanese families with autosomal-recessive type juvenile parkinsonism. AB - We present the clinical features of 17 patients from 12 Japanese families with familial juvenile parkinsonism suggesting autosomal-recessive inheritance (AR JP). Because the marriages of the parents in all but one family were consanguineous and all patients were only first generation, an autosomal recessive trait was strongly suspected. Analysis of the clinical features showed female predominance, mean age at onset 27.8 years, and slow progression. The symptoms of the parkinsonian triad (tremor, rigidity, and bradykinesia) were mild, but gait freezing, hyperreflexia, foot dystonia, and retropulsion were relatively prominent. A characteristic finding was amelioration of parkinsonian symptoms after sleep in all patients. Response to levodopa was satisfactory, but dopa-induced choreic limb dyskinesia and wearing-off phenomenon occurred frequently. Juvenile parkinsonism is a syndrome that encompasses several clinical entities. The similarity of clinical findings in these patients, and the differences from other types of parkinsonism, indicates that AR-JP is distinct clinical entity. PMID- 8710072 TI - Long-term tacrine (Cognex) treatment: effects on nursing home placement and mortality, Tacrine Study Group. AB - OBJECTIVE: To assess the possible association between tacrine (Cognex, manufactured by Parke-Davis, Morris Plains, NJ) dose and likelihood of nursing home placement (NHP) or death in patients with AD. DESIGN: A 30-week, randomized, double-blind, placebo-controlled, parallel-group multicenter clinical trial involving 663 patients, after which patients were treated openly and followed up a minimum of 2 years later. PATIENTS: At baseline, outpatients were at least 50 years of age, met criteria for probable AD, with baseline Mini-Mental State Examination scores between 10 and 26 (inclusive), were otherwise healthy, and had a caregiver who could provide assessments and ensure medication compliance. INTERVENTIONS: mandomized assignment to placebo or one of three ascending dosage regimens of tacrine over 30 weeks, followed by open label treatment for all patients who began the double-blind trial. OUTCOME MEASURES: NHP and death were examined using logistic regression. RESULTS: PATIENTS who remained on tacrine and were receiving doses > 80 mg/d or > 120 mg/d were less likely to have entered a nursing home than patients on lower doses (odds ratios > 2.7,2.8, respectively.) There was a trend for lower mortality for patients receiving > 120 mg/d (p = 0.063). CONCLUSIONS: Treatment with tacrine at doses > 80 mg/d was associated with a reduced likelihood of NHP. These data demonstrate that tacrine's 30-week effects on cognitive function and clinicians' global ratings may generalize to effects on a major milestone of AD. Future studies should attempt to replicate these findings prospectively. PMID- 8710073 TI - Category-specific difficulty naming with verbs in Alzheimer's disease. AB - We studied 20 patients with Alzheimer's disease (AD) on a picture-naming task consisting of frequency-matched pairs of nouns and verbs that were homophonic and homographic (e.g., paint). Intragroup comparisons revealed that verb naming is significantly more difficult for patients with AD than noun naming. An error analysis demonstrated that patients with AD produce significantly more semantic and descriptive errors for verbs than nouns. We correlated verb naming and noun naming with measures of grammatical comprehension, lexical retrieval, and visuoperceptual processing, but there were no selective effects for verbs compared with nouns. Differences in the mental representation of concepts underlying verbs and nouns may account, in part, for the relative difficulty naming with verbs in AD. PMID- 8710074 TI - Axonal channelopathies: an evolving concept in the pathogenesis of peripheral nerve disorders. AB - Abnormalities of peripheral nerve Na+ and K+ channels result in clinical manifestations unrelated to axonal degeneration or demyelination. Na+ channel blockade causes weakness and sensory loss associated with slowed conduction and decreased motor and sensory action potential amplitudes. K+ channel abnormalities result in high frequency repetitive action potentials (e.g., myokymia). Ion channel abnormalities may also play an important role in electrophysiologic changes seen in demyelinating disorders. PMID- 8710075 TI - Language comprehension profiles in Alzheimer's disease, multi-infarct dementia, and frontotemporal degeneration. AB - We assessed language functioning in 116 age-, education-, and severity-matched patients with the clinical diagnosis of Alzheimer's disease (AD), multi-infarct dementia (MID) due to small-vessel ischemic disease, or a frontotemporal form of degeneration (FD). Assessments of comprehension revealed that patients with AD are significantly impaired in their judgments of single word and picture meaning, whereas patients with FD had sentence comprehension difficulty due to impaired processing of grammatical phrase structure. Patients with MID did not differ from control subjects in their comprehension performance. Traditional aphasiologic measures did not distinguish between AD, MID, and FD. Selective patterns of comprehension difficulty in patients with different forms of dementia emphasize that language deficits cannot be explained entirely by the compromised memory associated with a progressive neurodegenerative illness. PMID- 8710076 TI - The apolipoprotein E epsilon 4 allele is associated with increased neuritic plaques and cerebral amyloid angiopathy in Alzheimer's disease and Lewy body variant. AB - OBJECTIVE: To determine the relationship between apolipoprotein E (apoE) genotype and neuropathologic lesions in Alzheimer's disease (AD) and Lewy body variant (LBV). DESIGN: Retrospective genetic-neuropathologic study of AD and LBV cases. The main neuropathologic outcome measures were modeled as a function of apoE genotype, neuropathologic diagnosis, and gender. Age at death and duration of symptom effects were controlled for by ANCOVA. PATIENTS: One hundred twenty-seven cases with neuropathologically diagnosed AD (n = 84) or LBV (n = 43). MAIN OUTCOME MEASURES: Quantitative scores of neuritic plaques (NPs), neurofibrillary tangles (NFTs), cerebral amyloid angiopathy (CAA) severity, and CAA prevalence were averaged across four brain regions: midfrontal, inferior parietal, superior temporal, and hippocampal. RESULTS: The apoE epsilon 4 allele was associated with increased NPs within both AD and LBV. The epsilon 4 allele was associated with an increased frequency of CAA in the AD and LBV groups combined groups combined and in and in LBV alone. While CAA severity and NETs were increased in the epsilon 4/4 homozygous case when AD and LBV were combined, there were no significant effects within AD or LBV alone. CONCLUSIONS: The apoE epsilon 4 allele is strongly associated with increased NPs, but not neocortical NFTs, in both AD and LBV. PMID- 8710077 TI - A newly identified polymorphism in the apolipoprotein E enhancer gene region is associated with Alzheimer's disease and strongly with the epsilon 4 allele. AB - Apolipoprotein E allele 4 (apoE epsilon 4) is a major risk factor for late-onset AD. Inheritance of this allele is associated with an earlier age of onset of dementia in individuals with AD. It is unknown whether other polymorphisms in the apoE gene may influence the effect of apoE epsilon 4 on AD. We screened portions of the promoter enhancer element and of the apoE receptor binding domain for other polymorphisms that could affect risk of AD. In particular, a C/G polymorphism at position +113 of the apoE mRNA in the apoE intron 1 enhancer element (IE1) has been recently identified. We found no other polymorphisms. We studied the relationship of the two alleles of the IE1 polymorphism with AD and found an apparent association between IE1 G and AD (n = 94; p = 0.0515). However, the IE1 G allele is also closely associated with apoE epsilon 4 (p < 0.0001). When the presence of apoE epsilon 4 is covaried, the association between the IE1 G allele and AD is no longer statistically significant (odds ratio = 1.29, 95% confidence interval: 0.44, 3.78). In contrast, epsilon 4 is still highly associated with AD when IE1 G is controlled for (odds ratio = 5.91, 95% confidence interval: 3.29, 10.63). Furthermore, there is no significant association between the age of onset of dementia and the inheritance of the G allele. We believe that the apparent association between IE1 G and AD is a consequence of the association between the epsilon 4 and IE1 G alleles. PMID- 8710078 TI - Abnormalities of torsional fast phase eye movements in unilateral rostral midbrain disease. AB - In a patient with a unilateral rostral midbrain lesion, three-dimensional scleral search coil eye movement recordings demonstrated slowing of ipsidirectional torsional fast phase eye movements without any abnormalities of torsional slow phases. On high-resolution MRI, the brainstem lesion localized to the area of the efferent pathways from the rostral interstitial nucleus of the medial longitudinal fasciculus (riMLF). This supports the experimental finding that unilateral inactivation of the riMLF results in a loss of ipsidirectional torsional fast phase eye movements and the hypothesis that there is lateralization of function throughout the torsional fast eye movement generating system. PMID- 8710079 TI - Speech timing in ataxic disorders: sentence production and rapid repetitive articulation. AB - We studied syllabic timing in patients with ataxia (10 with cerebellar atrophy, 6 with Friedreich's ataxia) under two conditions: in a "natural" sentence production context and in the context of a rapid syllable repetition task. The two tasks included comparable articulatory maneuvers. We measured syllable durations from the speech signal and analyzed variables describing average syllabic rate and within-trial variation of syllable durations. Among the observed measures, slowed syllable repetition was a particularly powerful predictor of the severity of dysarthric impairment. In sentence production, patients often performed at normal syllabic rates. Irregular pacing of syllable repetitions was frequent. Different patterns of between-articulator variation emerged in the two tasks. All patients except one were slower in rapid repetitive articulation than in sentence production. These data suggest that sentence production and rapid repetitive articulation are governed by basically different motor processes. The disproportionate slowing of ataxic patients in the repetitive task can be ascribed to adaptation to novel motor tasks being impaired in cerebellar disease. PMID- 8710080 TI - The world-wide burden of neurologic disease. PMID- 8710081 TI - Clinical evidence of genetic anticipation in adult-onset idiopathic dystonia. AB - Idiopathic dystonia occurs in both hereditary and sporadic forms. In this report, we studied the age of onset and family history of 260 patients (probands) with idiopathic adult-onset dystonia (IAD), cranial or cervical. The mean age at onset of these patients was (45.71 +/- 15.85) years. Forty-nine probands had a positive family history of dystonia or tremor in first- and second-degree relatives, and 7 had affected siblings only. The significance of tremor as a part of clinical manifestation of dystonia was evidenced by a high frequency of postural or action tremor in patients and relatives. Retrospectively, we examined the age of onset of dystonia (cervical or cranial) on successive generations in 49 families. Age of onset of clinical symptoms was earlier, by an average of 21.25 years, in the second generation than in the first generation. The mean age at onset of affected family members differed significantly between successive generations in these 49 families (p = 1.11 x 10(-8)). Our results suggest a tendency for earlier onset of dystonia and worsening of disease phenotype in succeeding generations in the same family. These findings are most compatible with genetic anticipation and suggest that an unstable trinucleotide repeat is most likely involved in adult-onset primary cranial or cervical dystonia. In addition, tremor as an integral part of dystonia needs further evaluation by molecular genetic studies. PMID- 8710082 TI - Consequence of nigrostriatal denervation and L-dopa therapy on the expression of glutamic acid decarboxylase messenger RNA in the pallidum. AB - To examine the consequences of nigrostriatal denervation and L-dopa treatment on the basal ganglia output system, we analyzed, by quantitative in situ hybridization, the messenger RNA coding for glutamic acid decarboxylase (Mr 67,000) (GAD67 mRNA) in pallidal cells from patients with Parkinson's disease (PD), monkeys rendered parkinsonian by 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP) receiving or not receiving L-dopa, and their respective control subjects. In MPTP-treated monkeys, the expression of GAD67 mRNA was increased in cells from the internal pallidum, and this effect was abolished by L dopa treatment. There were no differences in the levels of GAD67 mRNA between patients with PD, who were all treated with L-dopa, and control subjects. These results indicate that the level of GAD67 mRNA is increased in the cells of the internal pallidum after nigrostriatal dopaminergic denervation and that this increase can be reversed by L-dopa therapy. PMID- 8710083 TI - CYP2D6 allelic frequencies in young-onset Parkinson's disease. AB - Parkinson's disease (PD) is thought to develop as a result of interactions between genetic susceptibility factors and environmental exposures. One candidate gene is CYP2D6, which codes for the debrisoquine 4-hydroxylase cytochrome P450. Impairment of debrisoquine 4-hydroxylase activity has been associated with an increased risk of PD in patients with younger age at disease onset. Genotyping studies in patients with an older age at onset have reported modest increases in risk associated with the CYP2D6 B and A alleles; however, the risk for young onset PD has not been adequately evaluated. We designed a case-control study to investigate the role of nonfunctional CYP2D6 allelic risk factors for young-onset PD in a sizable patient population and compared the distributions of CYP2D6 genotypes between young-onset ( < or = 51 years) PD patients (n = 108) and controls (n = 236). In contrast with the results from genotyping studies conducted among patients with an older age at onset, there were no significant differences in CYP2D6 allelic frequencies between young-onset PD cases and controls. The frequency of the B allele was slightly lower in the young-onset PD cases than in the controls (0.14 versus 0.20) (X2 = 2.66, p = 0.10). The presence of one or more B alleles was not associated with an increased risk of young-onset PD (odds ratio 0.58; 95% CI 0.33 to 1.00), nor was the presence of one or more nonfunctional alleles (i.e., A, B, D, and D2) (odds ratio 0.68; 95% CI 0.41 to 1.13). This study suggests that the young-onset PD population may differ from the older-onset population with respect to risk factors. PMID- 8710084 TI - Spinal motoneuron excitability after acute spinal cord injury in humans. AB - BACKGROUND: Few studies in humans have assessed the ability of Ia afferent and antidromic motor volleys to activate motoneurons during spinal shock. Hence, little is known about the excitability state of the spinal motoneuron pool after acute spinal cord injury (SCI) in humans. METHODS: In 14 patients with acute SCI involving anatomic levels T10 and above, we performed clinical and electrophysiologic studies early after injury (within 24 hours in seven subjects) and on day 10, 20, and 30 postinjury. Maximal H:M ratios, F-wave persistence, and tendon tap T-reflexes were recorded. Sixteen normal subjects and eight chronic SCI patients served as control subjects. RESULTS: Ten of 14 patients had spinal shock (complete paralysis, loss of sensation, absent reflexes, and muscle hypotonia below the injury) at the time of initial evaluation. F-waves were absent in patients with spinal shock, reduced in persistence in patients with acute SCI without spinal shock, and normal in persistence in patients with chronic SCI. H-reflexes were absent or markedly suppressed in patients with spinal shock within 24 hours of injury but recovered to normal amplitudes within several days postinjury. This recovery occurred despite absence of F-waves that persisted for several weeks postinjury. Deep tendon reflexes were proportionally more depressed in spinal shock than were H-reflexes. All patients had elicitable H-reflexes for days or weeks before the development of clinical reflexes. CONCLUSIONS: Rostral cord injury causes postsynaptic changes (hyperpolarization) in caudal motoneurons. This hyperpolarization is a major physiologic derangement in spinal shock. The rise in H-reflex amplitude despite evidence of persistent hyperpolarization is due to enhanced transmission at Ia fiber-motoneuron connections below the SCI. Finally, the observation that the stretch reflex is proportionally more depressed than the H-reflex is consistent with fusimotor drive also being depressed after SCI. PMID- 8710085 TI - GM1b is a new member of antigen for serum antibody in Guillain-Barre syndrome. AB - Serum antibody from some patients with Guillain-Barre syndrome recognized an antigen of a minor component in human brain monosialoganglioside fraction. We purified that antigen, which migrated at a position slightly lower than that of GM1 on a thin-layer chromatogram (TLC), by using Iatrobeads column chromatography and preparative TLC. Structural analyses, including fast atom bombardment mass spectrometry, showed it to be GM1b. An enzyme-linked immunosorbent assay (ELISA) using purified GM1b showed that anti-GM1b antibody was present in 22 of 104 cases tested. No anti-GM1b antibody was present in the sera from control patients with other diseases or from the normal controls. Four sera recognized only GM1b among the 11 ganglioside antigens tested. The other 18 sera had antibodies to other antigens, most of which shared no terminal epitope with GM1b. Eight of nine sera samples with anti-GalNAc-GD1a antibody also had anti-GM1b antibody. Antibody to a minor monosialoganglioside, GM1b, was found to be a useful diagnostic marker for Guillain-Barre syndrome. Further study is needed to determine whether this antibody plays a role in the pathogenetic mechanism of the syndrome. PMID- 8710086 TI - Lethal infantile mitochondrial disease with isolated complex I deficiency in fibroblasts but with combined complex I and IV deficiencies in muscle. AB - A 2-month-old boy died of a lethal infantile mitochondrial disease with severe lactic acidosis and involvement of the CNS. Histochemical analysis of skeletal muscle showed that cytochrome c oxidase staining was lacking in all muscle fibers but was present in arterioles. Ragged red fibers were not seen, but some fibers showed excessive staining for succinate dehydrogenase. Biochemical analysis revealed a combined complex I and IV deficiency in skeletal muscle but only a complex I deficiency in his fibroblasts. Two-dimensional native SDS electrophoresis confirmed these enzymatic findings at the protein level. Analysis of mitochondrial translation products in fibroblasts revealed no abnormalities, and analysis of mitochondrial DNA in muscle showed no depletion, large-scale deletions, or frequently occurring point mutations. We conclude that this disease must have been the result of either a nuclear DNA mutation in a gene controlling the expression or assembly of both complex I and the muscle-specific isoform of complex IV or, alternatively, a heteroplasmic point mutation in a mitochondrial tRNA, which codon is used more often by mtDNA encoded subunits of complex I than by mtDNA encoded subunits of complex IV. A different degree of heteroplasmy in skeletal muscle and fibroblasts would then explain the curious heterogeneous tissue expression of defects in this patient. PMID- 8710087 TI - Decreased parvalbumin immunoreactivity in surviving Purkinje cells of patients with spinocerebellar ataxia-1. AB - The distribution of two calcium-binding proteins, calbindin D28k (CaBP) and parvalbumin (PV), was investigated by immunohistochemistry in the brains of three individuals dying of nonneurologic illness and three patients with spinocerebellar ataxia-1 (SCA-1). SCA-1 has recently been proven to be due to an unstable CAG repeat mutation on chromosome 6. In the cerebellum of control individuals the Purkinje cells showed strong immunoreactivity to CaBP. Other cells were CaBP-negative. Parvalbumin was highly localized to Purkinje, basket, stellate, and Golgi cells. All surviving Purkinje cells in SCA-1 were strongly immunoreactive to CaBP. The number of PV-immunoreactive Purkinje cells was markedly reduced in SCA-1. In addition, there was a significant decrease in the intensity of PV immunostaining within the individual Purkinje cells compared with controls. However, in the hippocampus, temporal cortex, and lateral geniculate scattered PV-positive neurons were seen in SCA-1 patients, similar to those in controls. The present results suggest that the decreased PV-immunoreactivity in the surviving Purkinje cells in SCA-1 may reflect biochemical alterations preceding Purkinje cell degeneration. PMID- 8710088 TI - Increased risk of dementia in mothers of Alzheimer's disease cases: evidence for maternal inheritance. AB - This study tests the hypothesis of maternal inheritance of AD in families of 118 subjects with this disorder enrolled in The Consortium to Establish a Registry for Alzheimer's Disease (CERAD). The parental generation included 24 subjects with dementia. Using the Cox proportional hazards model, we found the age adjusted mother-to-father relative risk to be 2.8 (95% CI, 1.1 to 7.7). Among a subset of 10 families with one affected parent and at least two affected siblings, the ratio of affected mothers-to-fathers was 9:1. These findings support recent studies that found a high mother-to-father ratio among affected parents of subjects with AD. Together, these results suggest maternal inheritance of AD and are consistent with several hypotheses regarding the genetic nature of AD. PMID- 8710089 TI - A preliminary study of apolipoprotein E genotype and psychiatric manifestations of Alzheimer's disease. AB - We evaluated the frequency of depression and psychosis in 46 patients with AD and 135 control subjects with the apolipoprotein (APO) E3/3 or E3/4 genotype. Patients with AD and the APOE3/4 genotype had a more than threefold increase in the signs of depression and psychosis when compared with either patients with the APOE3/3 genotype or to control subjects. Our preliminary study suggests that the phenotype of AD associated with the epsilon 4 allele is more likely to include psychiatric manifestations. PMID- 8710090 TI - Neuroimaging in the emergency patient presenting with seizure. PMID- 8710091 TI - Patient awareness of seizures. AB - In 31 consecutive patients who were admitted to an epilepsy monitoring unit, we prospectively determined whether the patients were aware of having seizures. On admission, all patients stated that they knew of at least some of their seizures. Eight of 23 with classifiable epileptic seizures recognized that they were occasionally unaware of their seizures. During telemetry, following full recovery of consciousness after each seizure, we asked the patients whether they had recently had a seizure. For control purposes, we asked the patients the same question at random times. Among patients with seizures, there were no false positive answers. Only 6 of 23 (26%) of the patients with epilepsy were always aware of their seizures, including complex partial and secondarily generalized events, and 7 of 23 (30%) were never aware of any seizures. Self-reporting of seizures was unreliable: Patients reporting the lowest baseline frequency of seizures had the highest fraction of unrecognized seizures. Seizure awareness was lowest for patients with temporal lobe foci, especially on the left side. Patients with primarily generalized epilepsy were more likely to be aware of tonic-clonic seizures than were patients with secondarily generalized partial seizures. All four patients with nonepileptic attacks believed that they always knew of their seizures, but only three of the four patients actually did always know. Unrecognized seizures are frequent and should be considered in patient management and in studies. PMID- 8710092 TI - Accuracy of reported family histories of essential tremor. AB - We studied the accuracy of reported family histories of essential tremor (ET) by questioning the patients in our clinic and subsequently by mail and phone. For individuals who continued to report a negative family history, we mailed a screening questionnaire to their first-degree relatives to further ascertain the presence of ET. On initial assessment, 67.7% of patients reported a positive family history of ET, but following all assessments, 96.0% of patients had a positive family history. We conclude that a negative family history of ET is often inaccurate, and that ET is primarily a hereditary disease. PMID- 8710093 TI - Susac's syndrome: neuropsychological characteristics in a young man. AB - A 29-year-old man developed vertigo, diplopia, confusion and spasticity. Brain biopsy revealed multifocal microinfarcts. Bilateral hearing loss and retinal microangiopathy were also noted. Deficits were stable years after onset. Repeated neuropsychological testing was suggestive of diffuse cerebral dysfunction, with likely prominent involvement of reciprocal diencephalic-cortical projections. PMID- 8710094 TI - Paraspinal muscle hematoma after electromyography. AB - There have been few reports of complications related to electromyography. Needle examination of certain muscles is sometimes avoided in patients taking anticoagulant agents, although no clear guidelines have been established. We describe a patient who was not receiving an anticoagulant and developed a large paraspinal muscle hematoma after routine electromyography. Subsequently, all patients who underwent paraspinal muscle electromyography and were diagnosed with radiculopathy at our institution over a 14-month period were reviewed. From this group, 17 patients were identified who had also underwent MRI of the appropriate spinal levels within 1 week after the needle examination. These images were reviewed for evidence of paraspinal muscle hematomas. Four small hematomas were identified in four different patients. None of these were radiologically significant compared with the large hematoma described in the case report. Radiologically apparent paraspinal hematomas after electromyography are an unusual complication of needle examination and do not appear to have any clinical significance. Nevertheless, the presence of these lesions justifies caution when considering electromyography of paraspinal and other deeper muscles in anticoagulated patients. PMID- 8710095 TI - Sodium channel accumulation in humans with painful neuromas. AB - Painful neuromas from 16 patients were examined using site-specific antisodium channel antibodies employed in immunocytochemical and radioimmunoassay methods. Normal sural nerves from six of these patients served as controls. Immunocytochemistry showed abnormal segmental accumulation of sodium channels within many axons in the neuromas. Dens immunolocalization was especially apparent within the axonal tips. Radioimmunoassay confirmed a significantly greater density of sodium channels in the neuromas as compared with the sural nerves. Thus, sodium channel accumulate abnormally within the axons of neuromas in humans. This alteration of the sodium channels may underlie the generation of axonal hyperexcitability and the resulting abnormal sensory phenomena (pain and paresthesias), which frequently occur after peripheral nerve injury. PMID- 8710096 TI - Traumatic occlusion of one limb of an intracranial arterial fenestration: an uncommon cause of stroke. AB - Three weeks after an automobile accident, a 35-year-old man experienced left throat and neck pain, numbness of the left face and tongue, dysphagia, left arm pain and weakness, and left miosis. At age 27, he had suffered an aneurysmal subarachnoid hemorrhage. Angiography at that time had also demonstrated a fenestration of the left intracranial vertebral artery. At the time of the second presentation, angiography showed that one of the limbs of the fenestration had become occluded. Although the vast majority of intracranial arterial fenestrations are asymptomatic, occlusion of one of the limbs of a fenestration may be the cause of stroke. PMID- 8710097 TI - US neurologists: attitudes on the US health care system. AB - Because the American Academy of Neurology (AAN) exists to represent the views and needs of neurologists, a mailed survey to a randomly selected sample of 520 US based, board-certified or board-eligible neurologist members was conducted in early 1995 to determine their attitudes in four key changing areas of the US health care system: (1) provision of neurologic care, (2) neurology workforce, (3) academic research, industry sponsorship, and pharmaceutical issues, and (4) health care delivery systems issues. Frequency distributions of degree of agreement and disagreement with 40 statements regarding issues in these four areas were tabulated for the entire group of 430 respondents (response rate = 83%). Associations between these attitudes and six demographic and practice variables (age, gender, board certification, practice type, degree of managed care, and geographical region) were evaluated in an exploratory analysis. The high response rate to the survey and the similarity in demographic and practice characteristics of respondents to the entire US-based, board-certified or eligible AAN membership support that the attitudes that were measured are likely to be representative of US neurologists, excluding resident trainees. The survey results show that neurologists are deeply concerned that patients with chronic neurologic disorders are not receiving all the neurologic services they need. Respondent show uncertainty about whether there is a current oversupply of neurologists but generally agree that there are too many neurology residency positions. Neurologists support industry sponsorship of academic medical center research and new drug development. With respect to many of the current mechanisms for health care cost containment, respondents indicate they have little confidence that these strategies are in the best interests of patients or physicians. The survey results confirm that there is no consensus among US neurologists on how to improve today's health care delivery system, and the results suggest that certain attitudes are related to neurologists' demographic and practice characteristics. Since today's health care market is volatile, regular assessments of neurologists' attitudes will be needed to assure that their views are understood and clearly represented by the AAN. PMID- 8710098 TI - Practice parameter: neuroimaging in the emergency patient presenting with seizure -summary statement. Quality Standards Subcommittee of the American Academy of Neurology in cooperation with American College of Emergency Physicians, American Association of Neurological Surgeons, and American Society of Neuroradiology. PMID- 8710099 TI - The lunatic and his seizure. PMID- 8710100 TI - Easy therapeutical management of sumatriptan-induced daily headache. PMID- 8710101 TI - Spinal air embolism resulting from a party game. PMID- 8710102 TI - Ictal radial and horizontal neglect. PMID- 8710103 TI - Aplastic anemia associated with ticlopidine. PMID- 8710104 TI - Cranial sinus thrombosis associated with essential thrombocythemia followed by heparin-associated thrombocytopenia. PMID- 8710105 TI - Cluster-tic syndrome. PMID- 8710106 TI - Psychogenic basilar migraine. PMID- 8710107 TI - Fluoxetine and seizures. PMID- 8710108 TI - N30 in PD. PMID- 8710109 TI - N30 in PD. PMID- 8710110 TI - Ocular palsy and CIPD. PMID- 8710111 TI - Cerebellar timing process. PMID- 8710112 TI - Complex partial status. PMID- 8710113 TI - Interferon ALFA. PMID- 8710114 TI - Brain death. PMID- 8710115 TI - Brain death. PMID- 8710116 TI - Cobalamin deficiency. PMID- 8710117 TI - Albino visual pathway. PMID- 8710118 TI - Reading epilepsy. PMID- 8710119 TI - SPECT in OPCA. PMID- 8710120 TI - Choosing the best needle for diagnostic lumbar puncture. AB - A new generation of Atraumatic (blunt tipped) needles now exists in addition to the traditional Quincke (cutting tip) needles for LP. We wished to identify the optimal size and type of needle that should be used for this technique. Requirements include rapid and accurate transduction of CSF pressure and adequate flow rate for CSF collection purposes while minimizing headache and other neurologic sequelae. Different types of available Atraumatic and Quincke needles were tested in two models that simulated normal and raised CSF pressures. The flow rates and the time required for an accurate transduction of CSF pressure onto a standard spinal manometer through each needle type was measured. Overall, Atraumatic-tipped needles compared favorably with similarly sized Quincke needles tested. The 20-gauge needles displayed suitable flow and pressure transduction characteristics. Some of the 22-gauge Atraumatic needles rapidly measured CSF pressure, but their flow rates were only suitable for small volume CSF collection. There is extensive literature to support that neurologic sequelae are reduced using Atraumatic needles. Diagnostic LP can be easily and accurately performed by using a large Atraumatic needle with the potential for considerable reduction in post-LP headache and related neurologic sequelae. PMID- 8710121 TI - Psychological factors in the diagnosis and pathogenesis of stiff-man syndrome. AB - Retrospective psychological evaluation of nine patients with stiff-man syndrome (SMS), seven of whom evidenced autoimmune disease, revealed a characteristic set of psychological symptoms or features: Major stressful life events preceded the development of permanent symptoms by 6 months or less (seven patients); transient motor symptoms occurred in emotionally distressing situations months or even years before the onset of a permanent motor deficit (five patients); after onset, similar situations specifically precipitated or augmented stiffness and spasms (five patients). We also found task-specific fear resembling agoraphobia (six patients) and loss or invalidation of one or both parents, or loss of home, in childhood (seven patients). Eight patients were initially misdiagnosed as having psychogenic movement disorder. We conclude that the common misdiagnosis of SMS as a psychogenic movement disorder is due to the compelling association of a set of salient psychological features, bizarre and fluctuating motor symptoms, and lack of approved neurologic signs. PMID- 8710122 TI - Relative absence of psychopathology in benign essential blepharospasm and hemifacial spasm. AB - We report on the psychopathology and illness-related changes of life in patients with benign essential blepharospasm (BEB) or hemifacial spasm (HFS). Fifty-six patients with BEB and 40 patients with HFS completed the SCL 90R, a screening instrument for psychiatric symptomatology, and the Freiburg Questionnaire for Dystonia (FQD), a questionnaire about psychosocial changes in subjects with movement disorders. In both BEB and HFS patients, the mean scores on all but one subscale of the SCL 90R remained within the double standard deviation of normal. In comparing BEB to HFS patients in illness-related changes of life, BEB patients were more severely disabled in all areas of life examined. Psychological distress in BEB, but not in HFS, correlated with physical disability and in particular with impairment of vision. PMID- 8710123 TI - Sumatriptan in clinical practice: a 2-year review of 453 migraine patients. AB - The long-term and within-patient consistency of the efficacy and tolerability of subcutaneous and oral sumatriptan in migraine was studied by retrospective survey of 2 years with mailed self-administered questionnaires in our neurology outpatient clinic. Subjects were migraine patients with or without aura (N = 869). We measured long-term use of sumatriptan and within-patient consistency and change over time of headache relief, headache recurrence, and chest symptoms after sumatriptan. The questionnaire was returned by 735 (85%) patients; 453 had used sumatriptan for nearly 28,000 attacks during 25 (median) months (92% > 1 year). Sumatriptan provided headache relief, mostly within 2 hours, in 85% of patients in at least two-thirds of their attacks. Of all patients, 75% experienced (usually multiple) headache recurrences in at least some and 40% in (nearly) all attacks. Median time to recurrence was 8 to 12 hours (range 1 to 30). Recurrence of aura was reported as well. Over 2 years, efficacy of sumatriptan had waned in 18% of patients (mainly because of increase in headache recurrence) and improved in 12% (mainly because of reduction of headache recurrence or adverse events or increase of headache relief); the number of monthly doses of sumatriptan had increased in 20%, reduced in 35%, and not changed in 45% of patients. Chest symptoms occurred in up to 58% of patients in at least some and in up to 42% of patients in all attacks, causing discontinuation of sumatriptan in 10%. In total, 111 patients (25%) discontinued sumatriptan mainly because of headache recurrence, adverse events, insufficient headache relief, or high price. In most patients, the effects of sumatriptan were consistent within subjects and over time. In most patients, sumatriptan provided rapid headache relief. Multiple headache recurrence was the major limitation. Chest symptoms were frequent but usually not serious if patients were forewarned. PMID- 8710124 TI - Variation in migraine prevalence by race. AB - PURPOSE: Estimates of migraine prevalence from African and Asian populations are lower than those observed in European and North American populations. To determine if these international differences reflect differences in cultural, environmental, or genetic factors, we compared the prevalence of migraine among Caucasians, African Americans, and Asian Americans in the United States. If genetic factors predominate, racial differences should persist in the United States. METHODS: In Baltimore County, Maryland, 12,328 individuals 18 to 65 years of age were selected by random-digit dialing and interviewed by telephone about their headaches. Migraine diagnoses were assigned using International Headache Society criteria. RESULTS: In women, migraine prevalence was significantly higher in Caucasians (20.4%) than in African (16.2%) or Asian (9.2%) Americans. A similar pattern was observed among men (8.6%, 7.2%, and 4.2%). African Americans were less likely to report nausea or vomiting with their attacks, but more likely to report higher levels of headache pain. In contrast, African Americans tended to be less disabled by their attacks than Caucasians. There were no statistically significant differences in associated features between Asian American and Caucasian migraineurs. CONCLUSIONS: In the United States, migraine prevalence is highest in Caucasians, followed by African Americans and Asian Americans. While differences in socioeconomic status, diet, and symptom reporting may contribute to differences in estimated prevalence, we suggest that race-related differences in genetic vulnerability to migraine are more likely to predominate as an explanatory factor. PMID- 8710125 TI - Serum prolactin concentrations are elevated after syncope. AB - The distinction between syncope and epileptic seizures is a common clinical diagnostic problem. Elevated serum prolactin (PRL) concentrations are used to help differentiate epileptic from nonepileptic attacks such as pseudoseizures. Reports of PRL concentrations following syncope have been variable. To determine whether PRL rises after syncope, we measured serum PRL concentrations during a 45 minute passive 60-degree head-up tilt in 21 patients with a history of near fainting or syncope. Head-up tilt triggered hypotension (mean arterial pressure 51 mm Hg, 95% CI = 45-57) with syncope in 11 patients. PRL concentrations were elevated ( > 19 ng/mL) and reached a maximum within the first 30 minutes after tilt-induced syncope in nine patients (PRL supine: 11 ng/mL, 95% CI = 7-15, vs. PRL after syncope: 52 ng/mL, 95% CI = 36-67; a greater than fourfold rise), while they remained unchanged in 10 patients who had a normal response to head-up tilt (PRL supine: 6 ng/mL, 95% CI = 5-8, vs. maximum PRL while upright: 8 ng/mL, 95% CI = 6-10). The findings indicate that elevated PRL concentrations are present after hypotensive syncope and are of little use in differentiating such syncope from epileptic seizures. PMID- 8710126 TI - Preictal pseudosleep: a new finding in psychogenic seizures. AB - BACKGROUND AND OBJECTIVE: The diagnosis of psychogenic seizures (pseudoseizures) may be difficult and usually rests on video-EEG monitoring. We observed that pseudoseizures often arise out of a state that we termed preictal pseudosleep. The objective of this study was to investigate this potential new sign in pseudoseizures. METHODS: We prospectively studied all patients who underwent noninvasive monitoring over a 10-month period. Patients were monitored for a duration of 1 to 19 days (mean 4.9), and were divided into two groups: pseudoseizures and epileptic seizures. Patients with both conditions were excluded. Preictal pseudosleep was defined as a state that resembled normal sleep by behavioral criteria alone (i.e. patient motionless and eyes closed), while EEG showed evidence of wakefulness (alpha rhythm, active EMG, and rapid eye movement). This state had to be sustained for at least 1 minute before clinical onset. RESULTS: Patients had 1 to 25 (mean 7) clinical events recorded. Preictal pseudosleep was seen in 10 of 18 patients with pseudoseizures and in none of 39 patients with epileptic seizures, yielding a sensitivity of 56% and a specificity of 100% for pseudoseizures. CONCLUSION: Because of a high specificity, preictal pseudosleep may be a useful adjunctive finding to support the diagnosis of pseudoseizures. PMID- 8710127 TI - Prognosis for total control of complex partial and secondarily generalized tonic clonic seizures. Department of Veterans Affairs Epilepsy Cooperative Studies No. 118 and No. 264 Group. AB - BACKGROUND: Two prospective observations of adults with symptomatic, localization related (partial) epilepsy included 1,102 patients in VA multicenter studies (VA 118 and VA-264). Analyses assessed the likelihood of remaining seizure free for 12 and 24 months after initiating adequate antiepileptic drug therapy. METHODS: Patients were grouped as having only secondarily generalized tonic-clonic seizures (GTC), only complex partial seizures (CPS), or both types (MIXED) at entry. The cumulative proportion of patients remaining seizure free with standard antiepileptic drug therapy was determined by actuarial life table methods. RESULTS: At 12 months, 70% and 61% of GTC patients (VA-118 and VA-264, respectively) had no further GTC; 53% and 50% of MIXED, predominantly GTC patients had no further GTC, 21% and 28% of CPS patients had no further CPS and 98% and 91% were seizure free for GTC; 32% and 35% of MIXED, predominantly CPS patients had no further CPS, and 62% and 51% of patients with MIXED seizure types remained seizure free for CPS for 12 months after enrollment. CONCLUSIONS: The overall prognosis for control of seizures of any type for 12 months was best for those who had only GTC at entry (55% and 48%), worst for those who had only CPS at entry (23% and 26%), and intermediate for those with MIXED seizures at entry (32% and 25%) (all p < 0.0001). Prognosis can be based on the predominant seizure type in patients with multiple types. PMID- 8710128 TI - The prognostic value of the electroencephalogram in antiepileptic drug withdrawal in partial epilepsies. AB - We evaluated the prognostic value of the EEG in 120 seizure-free epileptic patients (49 with complex partial seizures with or without episodic secondarily generalization [CPS], 20 with simple partial seizures with or without episodic secondarily generalization [SPS], 51 with only secondarily generalized seizures [PSG] during and after antiepileptic drug withdrawal. All patients had EEG examination before; during; and 3, 12, 24, and 36 months after drug withdrawal. Relapse rates were 45% in CPS, 100% in SPS, and 65% in PSG. Before reduction, 36 patients had epileptiform EEG and 69% relapsed; in the group with normal EEG, 60% relapsed. EEG worsened in 36 patients, 83% relapsed, whereas only 54% of patients with unchanged EEG relapsed. EEG during but not at the start of drug withdrawal has a prognostic value in partial epilepsy. PMID- 8710129 TI - Epileptic forced thinking from left frontal lesions. AB - Forced thinking is an incompletely understood and rarely described epileptic aura. We studied three patients with forced thinking from left frontal lesions, two neoplastic and one vascular. All three experienced repetitive, intrusive thoughts at the onset of seizures. Their forced thinking was associated with the desire to vocalize, orobuccal movements, and speech arrest. The episodes occurred with other ictal manifestations and responded to antiseizure therapy. These patients suggest that epileptic forced thinking is a heterogeneous phenomenon; forced thinking from left frontal lesions is a manifestation of expressive language and is distinct from experiential thoughts arising from temporal limbic foci. PMID- 8710130 TI - An assessment of nonconvulsive seizures in the intensive care unit using continuous EEG monitoring: an investigation of variables associated with mortality. AB - Of 49 patients with nonconvulsive seizures studied with continuous EEG monitoring, the overall mortality was 33% (16/49). Of the 23 patients with nonconvulsive status epilepticus (NCSE), 13 died (mortality 57%). Individual variables significantly associated with mortality were age, presence of NCSE, seizure duration, hospital and NICU length of stay, and delay to diagnosis and etiology (acute illness versus remote symptomatic). With multivariate logistic regression, only seizure duration (p = 0.0057, OR = 1.131/hour) and delay to diagnosis (p = 0.0351, OR = 1.039/hour) were associated with increased mortality. Acute symptomatic cases could not be adequately classified as either absence, simple, or complex partial status epilepticus when the impairment of consciousness arose form the initial illness. Current classifications of status epilepticus are inadequate for such cases. PMID- 8710131 TI - EEG-triggered echo-planar functional MRI in epilepsy. AB - We investigated whether: (1) EEG recordings could be successfully performed in an MRI imager, (2) subclinical epileptic discharges could be used to trigger ultrafast functional MRI images, (3) artifact-free functional MRI images could be obtained while the patient was having the EEG monitored, and (4) the functional MRI images so obtained would show focal signal increases in relation to epileptic discharges. We report our results in two patients who showed focally higher signal intensity, reflective of increased local blood flow, in ultrafast functional MRI timed to epileptic discharges recorded while the patients were in the imager and compared with images not associated with discharges. One patient showed a focal increase despite a clinical and EEG history of generalized discharges. This approach may have the potential to identify brain regions activated during brief focal epileptic discharges. PMID- 8710132 TI - Isolated vertigo as a manifestation of vertebrobasilar ischemia. AB - OBJECTIVE: We sought to demonstrate that isolated episodes of vertigo can be the only manifestation of vertebrobasilar ischemia. BACKGROUND: Isolated persistent vertigo is classically ascribed to labyrinthine disorders and is only rarely considered to reflect vertebrobasilar ischemia. METHODS: We retrospectively analyzed all of the records of the Saint Louis University Stroke Registry between January 1, 1992 and September 1, 1993. We set out to identify those patients discharged with a diagnosis of transient ischemic attack (TIA) in the vertebrobasilar system. We reviewed their clinical records and the results of their diagnostic studies. RESULTS: We screened 600 admissions and found 29 patients with vertebrobasilar circulation TIAs. Of these, five men and one woman had episodic vertigo for at least 4 weeks as their only presenting symptom. All six patients had one of two abnormal patterns on magnetic resonance angiography (MRA): focal basilar stenosis or widespread vertebrobasilar slow flow. In three patients, the MRA findings were confirmed by cerebral angiography. Five patients were treated with warfarin and one with aspirin. Two patients developed brainstem infarctions, one of them fatal. CONCLUSIONS: Isolated vertigo can be the only manifestation of vertebrobasilar ischemia. Its frequency may be underestimated in clinical practice. Noninvasive testing is helpful both for diagnosis and follow up. PMID- 8710133 TI - Childhood chronic inflammatory demyelinating neuropathies: clinical course and long-term follow-up. AB - Chronic inflammatory demyelinating neuropathy (CIDP) is a rare disease in childhood. We reviewed the clinical characteristics, response to therapy, and long-term prognosis in 13 children (1.5 to 16 years of age) diagnosed with CIDP at Washington University Medical Center, St. Louis, and the Royal Children's Hospital, Melbourne, Australia, between 1979 and 1994. The most common presenting symptom (in 11/13 [85%]) was lower extremity weakness associated with difficulty in walking. Preceding events within 1 months of onset, mostly intercurrent infections or vaccinations, occurred in seven children (54%). The disease was monophasic in three children (23%). One relapse occurred in four (30%) and multiple relapses in six (46%). All patients had at least short-term response to steroids. Three children (23%) recovered completely during the first year. Ten children (77%) had residual weakness after an average follow-up of 6 years. There seems to be two populations of children with CIDP. One subgroup, with a favorable prognosis, progressed to peak disability over less than 3 months; these children often have a monophasic course with complete resolution of symptoms and signs and withdrawal from all medications by 1 year after onset. A second subgroup progressed for 3 months or longer; these children all required substantial does of prednisone for prolonged periods and had considerable long-term morbidity with persistent weakness. PMID- 8710134 TI - [Doppler evaluation of diastolic function in hypertrophic cardiomyopathy]. AB - Diastolic function may play a significant role in patients affected by hypertrophic cardiomyopathy, because abnormalities in diastolic function, even in presence of a normal systolic function, may determine the clinical features of the disease. Doppler ecocardiography, using a non-invasive and not highly expensive method, easily allows to analize the indexes of diastolic function, as soon as some morpho-functional parameters, represented by extent and localization of the myocardial hypertrophy and presence or absence of obstruction in the left ventricular outflow, which have always been considered relevant in the prognosis together with the typical arrhythmic abnormalities of the disease. The aim of this study was to investigate the behaviour of Doppler ecocardiographic parameters of diastolic function in 38 patients with hypertrophic cardiomyopathy and to analize whether they might be correlated with the morpho-functional patterns and clinical features of the disease, represented by the NYHA functional class and occurrence of ventricular tachycardia during 48 hour ambulatory electrocardiographic monitoring. Diastolic function abnormalities, although occurring in most patients, do not seem to be related with clinical conditions and/or with the typical morpho-functional patterns of the disease; however, an increase in the left atrial size together with a specific increase in the rate of deceleration of flow velocity in early diastole that were detected in patients with ventricular tachycardia, by suggesting a relation between diastolic dysfunction and arrhythmogenic substrate, remarks the importance of the analysis of diastolic function for a better prognostic evaluation of the hypertrophic cardiomyopathy. PMID- 8710135 TI - Carotid endarterectomy and myocardial revascularization. A single stage procedure. AB - Carotid endarterectomy (CEA) and myocardial revascularization can be performed in a single procedure, performing CEA before or during cardio-pulmonary by-pass (CPB), or using a double stage approach. Over a 4 year period, 17 patients underwent CEA and coronary artery by-pass (CAB) with a single stage procedure. Fourteen patients (82.3%) were male, 3 (17.6%) were female. The mean age was 66.3 +/- 7.07. One patient (5.8%) had a previous neurological event (stroke); 5 patients (29.4%) had a previous transient ischemic attacks (TIA). The indications for the combined operations were CAD associated to unilateral internal carotid stenosis greater than 70% or 50% when symptomatic. In all patients CEA was performed after median sternotomy and heparinization, during CPB, with moderate hypotermia (30%C), performing CEA successively. One patient (5.8%) died of acute heart failure secondary to mediastinits. Minor neurological complications were present in 2 patients (11.7%) with signs of cerebral oedema. Myocardial infarction and late neurological deficit did not occur in any patient. We conclude that it is important, in the preoperative assessment of every patients with CAD, the screening for concomitant carotid vascular diseases, in order to avoid neurological complications during CPB, treating the two different diseases with a single stage approach, if carotid stenosis is greater than 70% or greater than 50% when symptomatic. PMID- 8710136 TI - Improvement of postoperative graft patency rate for coronary revascularization. AB - For coronary artery revascularization, the long term patency rate of internal thoracic artery (ITA) is excellent and arterial conduit for coronary bypass grafting (CABG) has been actively pursued. Application of the right gastroepiploic artery (GEA) is on the increase but, the patency of the GEA graft is more technically demanding than that of ITA. To improve early postoperative graft patency for coronary artery bypass grafting (CABG) for arterial graft, we modified the technique for anastomosis and demonstrated the patency by postoperative angiogram in 26 consecutive patients receiving CABG. The graft was anastomosed to the coronary artery using two 8-0. Prolene sutures for arterial graft and 7-0 Prolene sutures for saphenous vein (SV) graft. The heal and toe sides of the graft were sutured separately by the parachute technique (double parachute technique), followed by running suturing and typing at bilateral sides. The number of distal anastomoses was 3.4/patient and 44 arterial grafts and 40 SV grafts were used for coronary revascularization. Arterial grafts consisted of 33 ITAs, 9 GEAs, and two inferior epigastric arteries. The angiograms taken 10 to 14 days after operation demonstrated 100% patency rate of arterial grafts and 39 out of 40 SV grafts were patent. The overall patency rate in the early postoperative period was 98.8% The "double parachute" technique for CABG is highly accurate anastomosis with good visualization and the patency rates for all kinds of grafts were found to have improved. PMID- 8710137 TI - Left ventricular performance after intravenous infusion of captopril in patients with congestive heart failure. AB - BACKGROUND: Although oral administration of captopril, an angiotensin-converting enzyme inhibitor, is effective for the treatment of congestive heart failure (CHF), the effect of its intravenous (iv) administration is not well known. METHODS AND RESULTS: Ten patients (age range 48-72 years), with CHF belonging to the second and third NYHA class, were given an iv bolus of 25 mg of captopril. Before and 30 minutes after the infusion of captopril, a number of parameters of the left ventricular function were evaluated by echocardiography IREX 3 M-B Mode. Eight patients showed a significant improvement of left ventricular performance indices. In fact, the ejection fraction (13.8%, p < 0.05), the cardiac output (24%, p < 0.001), the circumferential shortness fraction (29.9%, p < 0.05), and the fraction shortening (16.0%, p < 0.005) increased significantly, whereas the end-systolic diameter (21%, p < 0.001), the endsystolic stress (23.8%, p < 0.01) and the left ventricle ejection time (4.8%, p < 0.05) decreased significantly. Systolic and diastolic blood pressure values also underwent a significant reduction by 17% and 11% (p < 0.01 and p < 0.05 respectively). No evident correlation between the improvement of the left ventricular function and the basal renin rates was noticed. CONCLUSIONS: A significant improvement of parietal kinesis was observed especially in those segments which showed movement abnormalities (hypokinesia and akinesia) and in many cases this was detected by M B Mode echocardiography. Our findings may be the result of the following factors: 1) reduction of parietal stress; 2) increased district coronary flow; 3) inhibition of tissue renin-angiotensin-aldosterone system; and 4) "scavenging" action exerted by the SH group of captopril. PMID- 8710138 TI - Tissue protection by anti-ischemic drugs. AB - Ischemia followed by reperfusion is associated with typical microvascular disturbances characterized by capillary perfusion break-down (no-reflow), accumulation and adhesion of leukocytes to the microvascular endothelium (reflow paradox) and impairment of endothelial barrier function, indicating the onset of cellular and/or organ dysfunction. Therapeutic approaches aiming at the prevention of microvascular failure elicited by ischemia/reperfusion, have to be devided into measures interfering with 1) the insult induced during the ischemic period and its consequences from cellular energy depletion and 2) the microvascular disturbances induced during the reperfusion/reoxygenation period. Using various microcirculation models in hamster and/or mouse allowing for chronic visualization of the microcirculation in striated muscle and skin, different therapeutic strategies have been investigated in our laboratory to protect the tissue from the manifestation of postischemic microvascular disturbances. The results are discussed with respect to the potential mode of action of the various therapeutic strategies performed. PMID- 8710139 TI - [Pulmonary thromboembolism. A clinical case with unusual presentation]. AB - The authors describe a rare case of pulmonary thromboembolism with unusual clinical findings and emphasized the large difficulty encountered in formuling a correct diagnosis in a reasonable time. A man, 60 years old, was admitted to a Medical Division of our hospital for the appearance of chest pain and epigastric pain during effort in the last year. He smoked 20 cigarettes a day and drank wine (1 or 2 litres a day). He was affected by hypercholesterolemia and in the past reported relapsed thrombophlebitis in the left leg. Four years before admission to our hospital he underwent large and small left saphenectomy. He had no cardiac events in the past. After a non significant exercise stress test the patient was treated with nitrates and asa and was discharged from the hospital. At home the symptoms increased and after 8 months the patient was admitted again to the Cardiologic Division of the hospital. At admission he reported dyspnea and chest pain at rest, not only during effort and the ECG showed negative T waves in anterior and inferior leads. Intravenous heparine, nitrates and calcium antagonists stabilized the clinical picture. The following examinations revealed: reduction of the T wave negativity at the ECG registered during chest pain; mild enlargement of the heart at the chest roentgenogram; normal value of the left ventricle and apical and midseptal by ipokinesia at the transthoracic echocardiogram; normal coronary artery at the coronary arteriography. "Vasospastic angina" was diagnosed and the patient was discharged after 20 days, asymptomatic. After 15 days he returned to the hospital again for chest pain, dyspnea, hypotension and syncope despite therapy. At physical examination he showed a painful left tibio-tarsal tumefaction, an increased and splitting second heart sound in the pulmonary area and a systolic murmur in the third and fourth left interspace. The ECG showed a severe anterior ischemia, while a new transthoracic echocardiogram revealed a considerable dilatation of the right atrium, right ventricle and the main pulmonary artery with severe tricuspid regurgitation and pulmonary hypertension (mean PAP about 50 mmHg). The following pulmonary perfusion scintigraphy confirmed the diagnosis of pulmonary embolism and the selective right and left pulmonary arteriography exhibited multiple thrombi and large intravascular filling defects. The right heart catheterization confirmed a chronic precapillary pulmonary hypertension (mean PAP = 55 mmHg). About 24 hours after these examinations the patient died because of a cardiac arrest with electromechanical dissociation. Pulmonary thromboembolism is a potentially fatal disease characterized by a largely variable clinical presentation. Frequently pulmonary embolism diagnosis is difficult especially when clinical findings are unusual. In the case observed the "typical" chest and epigastric pains associated with the electrocardiographic findings directed diagnosis towards myocardial ischemia. Also after the coronary arteriography that showed normal coronary artery, the erroneous diagnosis persisted. Pulmonary embolism was correctly diagnosed too late to begin an effective therapy. These unusual clinical findings and diagnostic mistakes are stressed and critically reviewed in the article. PMID- 8710140 TI - [Reflex syncope as a preliminary symptom of a rhinopharyngeal lymphoma. A case report]. AB - The authors report a 57 year old patient affected by rhinopharyngeal NH lymphoma that appeared as a parapharyngeal space lesions-syncope syndrome. The activation of a cardioinhybitory reflex by the stimulation of the glossopharyngeal nerve seems to be the pathogenetic basis for his syndrome. PMID- 8710141 TI - [Hemoperitoneum from post-traumatic ovarian cystoma. Videolaparoscopic treatment]. AB - Personal experience in a case of ovarian cystoma (misunderstood to the patient herself) broken consequently to a car trauma and cause of an intracystic angiorrhagy and hemoperitoneum is presented. The advantages of videolaparoscopic technique both for the diagnosis formulation and for the surgical treatment are underlined and the use of this new technique is suggested due to its advantages such as quickness, mini-invasiveness and post-operation comfort. PMID- 8710142 TI - [Efferent loop obstruction by a phytobezoar after gastrectomy]. AB - The authors present one case of intestinal obstruction due to phytobezoar of the efferent loop, in a previously undergone gastric surgery patient. They report, in the light of bezoar's peculiar location, that is usually it found in the stomach or small bowel. PMID- 8710143 TI - [Bilateral thrombosis of a femoral pseudoaneurysm]. AB - One case has been described of acute thrombosis of bilateral femoral anastomotic pseudoaneurysm. The clinical finding of bilateral acute ischemia of the lower limbs required urgent surgical treatment. The operative technique consisted of excision of anastomotic aneurysms and replacement by a new segment of prosthesis with an end-to-end anastomosis to the deep femoral artery. Pathogenesis of the pseudoaneurysms at the femoral anastomosis generally recognized numerous factors such as mechanical, graft or suture defects, hypertension, wound complications. Recognition of femoral anastomotic aneurysms is usually simple, when a pulsative mass is noted. In such a case (reported) of thrombosis of bilateral femoral pseudoaneurysms, preoperative diagnosis was more difficult. Anastomotic aneurysms of little size that occur later after original intervention require observation by echography and angiodinography. When rapid enlargement arises, urgent surgical treatment is required, before rupture or thrombosis. The surgical intervention consists of excision of the anastomotic aneurysm and replacement by a new segment of prosthesis between the prosthesis and the common or deep femoral artery. If the reconstruction at the level of femoral artery is not possible, the anastomosis is performed more distally, at the level of popliteal artery. The results are affected by the degree of urgency of surgery, with significant difference in favour of the patients Who underwent elective procedures. PMID- 8710144 TI - [Aneurysms of the popliteal artery. A clinical case and revision of the literature]. AB - The authors report their experience about one case of popliteal artery aneurysm. The patient was treated with aneurysm resection and dacron by-pass graft. The good surgical results and high risk of natural complications of popliteal aneurysms suggest that operation in the asymptomatic stage should always be attempted. PMID- 8710145 TI - [Frey's syndrome: physiopathology and medical therapy]. AB - Frey syndrome (gustatory sweating, auricolotemporal syndrome) is a complication of parotidectomy, probably caused by misdirection of regenerating fibers in the auricolotemporal nerve. The authors review the pathophysiology and describe the treatment used in this entity. PMID- 8710146 TI - [Diagnostic and therapeutic approach to incidentally detected adrenal tumors]. AB - The authors report their experience relative to 12 cases of adrenal "incidentaloma" surgically treated. All masses removed had a main diameter larger than 3 cm. In all patients an anterior median transperitoneal incision was performed. They didn't record postoperative mortality and the postoperative morbidity rate was 15%. In 6 cases a nonfunctioning cortical adenoma was diagnosed. No malignant neoplasm was detected in their series. PMID- 8710147 TI - [Multiple primary malignant neoplasms]. AB - Multiple primary malignant neoplasms (MPMN) are tumors that arise independently in the same organ or tissues and different organs; they may present simultaneously (synchronous) or at least 6 months later (metachronous). The authors report the case observed: 20 multiple neoplasms in 5 years (5.8% of the tumors). In the light of the international literature they analyze some aspects of the problem, particularly with regard to the classification, etiopathogenesis, neoplasm associations and possible genetic character for some of there. PMID- 8710148 TI - [Treatment of large bowel obstruction. Experience with intra-operative wash-out of the colon]. AB - The treatment of colorectal obstructions is a surgical problem. The surgeon can choose between primary resection with anastomosis and the staged operations. The one stage procedures need colon decompression or intraoperative colonic lavage. In our experience between 1990 and 1993, 23 patients required an emergency intervention for colon obstruction; between them 13 patients were affected by a left colonic obstruction and were treated with a staged procedure (like Hartman operation) in 9 cases and with intraoperative colonic wash-out with primary anastomosis in 4 cases. The last group had a good postoperative course without an increased incidence of anastomotic leakage (no one in our limited experience). Compared with staged surgery, immediate resection and anastomosis had significant advantages for the patients because: 1) the quality of the life is better (absence of colonstomy); 2) the cumulative hospitalization is reduced (15 days vs 32 days); 3) there is a reduction in operative risk and in the cumulative intra- and postoperative immunodepression. The correct evaluation of the effect on the long-term survival of these factors needs larger series and of longer follow-up. PMID- 8710149 TI - [Complications of ileal lymphoma]. AB - Non-Hodgkin lymphoma of the ileum accounts for some 3% of all extranodal onset lymphoma and 20% of gastrointestinal lymphoma given that the ileum is more frequently affected than the jejunum and duodenum. The large majority of primary extranodal lymphomas present a diffuse histological structure and in particular involve the cervico-fascial and gastrointestinal regions. Moreover, it is not uncommon to find an association between gastroenteric involvement and Waldeyer's ring (cervico-fascial region). Primary intestinal involvement may not present specific symptoms and remain silent for some time. It is manifested by the onset of complications caused by occlusion and perforation. Two cases of ileal lymphoma were treated at the Institute of Emergency Surgery of Catania University between 1992 and 1993. They were complicated by intestinal perforation and occlusion respectively. Both patients underwent emergency intestinal resection. Surgery represents the elective treatment for primary forms, followed by polychemotherapy and radiotherapy. Prognosis depends on the spread of disease and the hystotype. The administration of NTP and somatopstatin resulted in a shorter postoperative period with fewer surgical complications. PMID- 8710150 TI - [Video-assisted thoracoscopy in the treatment of recurrent pneumothorax]. AB - Between April 1992 and May 1994, 45 patients with recurrent spontaneous pneumothorax underwent videoassisted thoracoscopy (group I). The mean chest tube duration, the length of hospital stay, the use of parenteral narcotics, the complications and the follow-up were analyzed and compared to the same data of a group of 21 patients previously treated by open approach between January 1991 and March 1993 (group II). Average age, sex and surgical indications distribution were comparable (group I: 36 males, 9 females, mean age 31.7 years; group II: 17 males, 4 females, mean age 31.5 years). Mean chest tube duration was lower in group I (group I 4.3 days vs group II 7.2 days), as was mean hospital stay (group I 4.6 days vs group II 10.3 days) and the necessity of parenteral narcotics for pain relief (group I 11% vs group II 66% of patients). No episodes of relapsing pneumothorax occurred in either group of patients after a mean follow-up of 12.4 months (range from 1-24 months) for group I and 30 months (range from 24-36 months) for group II. The incidence of minor complications was less in group I (4.4%) than group II (23.8%). Our early results in the treatment of recurrent spontaneous pneumothorax by videoassisted thoracoscopy have been encouraging and the merits of this approach make it preferable to thoracotomy. PMID- 8710152 TI - [Transversal cervicotomy in thyroid surgery. Indications for its enlargement]. AB - Surgical therapy of thyroid diseases utilizes a collar-shaped transverse cervicotomic access. However, this access can be insufficient for a complete execution of operating plan. The authors suggest sternum thoracic and laterocervical enlargement of the operative filed. These two enlargements present several advantages and don't expose the patient to greater postoperative complications. Moreover, the surgeon can operate with higher safety in controlling inferior instestinal peduncolar and recurrent nerve. In case of latero-cervical access a complete dissection of neck is also possible. PMID- 8710151 TI - [Pleural drainage vs video-assisted minimally invasive surgery in the treatment of spontaneous pneumothorax. A retrospective study]. AB - The most frequent cause of spontaneous pneumothorax is bleb's disease of the lung. Considering that, pnx s. has a remarkable bent for relapsing, we think that it's necessary an aggressive treatment to resolve it. That's because pleural drainage is useful for a symptomatic resolution, VATS for an etiologycal one. To value the real efficacy of the treatment of VATS, we have effected a retrospective research between two classes of patients suffering from pnx s. admitted in our Institute from 1987 until 1991, one treated with pleural drainage (class A, 11 patients) and the other with VATS (class B, 13 patients). 1) Patients treated with pleural drainage were nearly double compared to class B. 2) In class A, the mean value of drainages were nearly double compared to class B. 3) Class A had a number of relapses 10 times more compared to B. 4) Considering the period of hospitalization, class A had a value nearly triple compared to B. 5) We've noticed that the mean value of the cost for episode of pnx s. in class A was more than 15% compared to class B and the cost for patient in class A was nearly double than in class B. We can affirm that, for the treatment of pnx s., the method of VATS rappresents a real success. We're passed from the "symptomatological treatment" of it to an aetiological one, joined to the prophylaxis of the relapse. PMID- 8710153 TI - [Cervico-mediastinal goiters. Clinical aspects]. AB - Among 660 cases of thyroid pathology diagnosed by the 2nd Institute of Clinical Surgery, Policlinico San Matteo, IRCCS, Pavia, during the period 1984 to 1993, 34 cases of cervical-mediastinal goiter were observed with an incidence of 5%. After a careful analysis of the main contributions to the study of cervical-mediastinal goiters, paying special attention to their classification, pathogenesis, methods of growth, and unique symptomatic manifestations, the authors focus on current radiological and instrumental studies, which are widely regarded as being useful for the diagnosis of the site and nature of this disease. They review the routes for aggressive surgery and appropriate techniques for the exeresis of the mediastinal mass. Having empasised the absolute indication for surgery the authors recommend quasi-total or total thyroidectomy as the elective form of surgery. PMID- 8710154 TI - [Persistent varicocele: experience with prophilaxis and microsurgical treatment]. AB - Persisting varicocele is still a rather hard clinical and therapeutical problem requiring a good level of surgical and microsurgical capability. In order to prevent and to treat postsurgical persistence, our trend is to resort to a correct and rational etiopathogenetical framing allowing us to choose the most appropriate surgical treatment. Relying on such procedure we perform all sorts of traditional and microsurgical interventions. We can say that the microsurgical techniques are the most appropriate ones to solve the problems of the second surgical intervention. PMID- 8710156 TI - Joint action nephrology in Eastern Europe. International Society of Nephrology, European Renal Association (EDTA/ERA), and Kuratorium der Gesellschaft fur Nephrologie. PMID- 8710155 TI - [Bilateral and subclinical varicocele: clinical experience]. AB - The authors report their experience in clinical and instrumental diagnosis of primary varicocele, with regard to bilateral and subclinical forms. 54 patients affected by varicocele were evaluated comparing the results of clinical examination with those coming from scrotal ultrasonography and Doppler flowmetry of spermatic cords. Bilateral varicocele was found in 26 cases (48.2%); subclinical varicocele, monolateral or bilateral, was evidenced in 25 cases (46.3%). On the basis of this study clinical examination seems not to be ideal for the diagnosis of varicocele, particularly in view of a surgical or percutaneous treatment aimed at complete resolution of venous spermatic reflux. PMID- 8710157 TI - Proteinuria and blood pressure as causal components of progression to end-stage renal failure. Northern Italian Cooperative Study Group. AB - AIMS: To identify the prognostic factors possibly related to end-stage renal failure development. SUBJECTS AND METHODS: The prognostic factors affecting chronic renal failure progression were analysed in 456 patients who had participated in a formal, multicentre, prospective randomized trial aimed at verifying the role of protein restriction in slowing down or halting the progression of chronic renal failure. The 24-month follow-up foreseen by the protocol was completed by 311 patients and 69 reached an end-point. An inductive analysis on patient survival was made by using the Cox proportional hazard regression model, using a stepwise procedure in order to select only those factors which are significantly associated with survival. For each individual risk factor, a univariate descriptive analysis of survival was performed using the Kaplan-Meier technique. RESULTS: Underlying nephropathy, baseline plasma creatinine, proteinuria, and plasma calcium were all shown to be related to end stage renal failure onset. Hypertensive patients (mean blood pressure > 107 mmHg) had a worst cumulative renal survival but the degree of proteinuria was even more important as a prognostic factor of renal death than hypertension. The cumulative renal survival of patients whose proteinuria decreased during the trial follow-up was better than those of patients without changes. However, the interaction between baseline lying mean blood pressure and proteinuria was not significant. CONCLUSIONS: Only primary renal disease and proteinuria were related to renal survival, being baseline plasma creatinine confounding factor. By blocking the possible causal role of proteinuria and hypertension, end-stage renal failure could be prevented in a significant percentage of patients. PMID- 8710159 TI - Perirenal capsula adiposa--is it a sign indicating diabetes mellitus? PMID- 8710158 TI - Experience with zinc protoporphyrin as a marker of endogenous iron availability in chronic haemodialysis patients. AB - BACKGROUND: Ferritin and the percentage of transferrin saturation (TS) are established parameters with which to evaluate endogenous iron availability during treatment of renal anaemia with recombinant human erythropoietin (rHuEpo). Zinc protoporphyrin (ZPP) has been proposed as another valid marker in this setting. METHODS: We determined the following parameters in 127 patients, including 117 haemodialysis patients: haemoglobin, erythrocytes, haematocrit, mean corpuscular volume (MCV), iron, ferritin, transferrin saturation and ZPP. Of the patients treated in a cross-sectional study, 38.5% were treated with rHuEpo; 30.7% with intravenous iron; and 13.6% with intravenous iron and rHuEpo simultaneously. Median ferritin was 304 ng/ml and median transferrin saturation was 21.2%. RESULTS: Including cases with manifest storage iron deficiency, a concordant elevated ZPP ( > 40 mumol/mol haem) and a decreased transferrin saturation ( < 20%) were found in 23 of our dialysis patients (19.6%) while 55 cases (47%) were classified as concordantly negative. However, as many as 39 cases (33.3%) showed discrepant results: in 16 cases (13.6%) ZPP was elevated but transferrin saturation was in the normal range, while in 23 cases (19.6%) the opposite results were observed. CONCLUSIONS: We conclude that beyond the border of manifest storage iron deficiency, defined as a ferritin < 30 ng/ml in male and < 15 ng/ml in female patients, ZPP and TS cover different ranges of functional iron deficiency which is reflected in the lack of a correlation of ZPP to any other of the above-mentioned parameters. Our data suggest that a TS < 20% as a diagnostic, and thus intervention, criterion in the evaluation of functional iron deficiency and iron substitution beyond manifest storage iron deficiency might result in overestimation of iron requirements. It remains to be shown in a longitudinal study, also reflecting the course of haemoglobin and the mean rHuEpo dose, whether ZPP or TS is the more appropriate parameter in the evaluation of functional iron availability. PMID- 8710160 TI - August Heidland (Wurzburg)--recipient of the Franz Volhard Medal. PMID- 8710161 TI - Proton modulation of outward K+ currents in interferon-gamma-activated microglia. AB - Whole-cell outward potassium currents (IK) were measured in interferon (IFN) gamma-activated cultured murine microglial cells. Acidification of the external milieu moved the threshold of activation of IK in a depolarizing direction, while alkalinization showed the opposite effect. A shift of more than 20 mV of the steady-state activation and inactivation curves of IK in hyperpolarizing direction was measured when pH was changed from 5.8 to 7.8. The time-dependent inactivation of IK was slower when superfusing cells with acid solutions than with alkaline ones. In contrast, variations in the pH of the intracellular solution did not alter kinetics of IK. However, alkalinization of the internal solution from a pH of 5.8 to 7.8 led to a two-fold increase in the current density of IK. PMID- 8710162 TI - Differentiation of cholinergic cells in the rat gut during pre- and postnatal life. AB - In the present work we studied the distribution and number of cholinergic neurons in the rat stomach, ileum and colon starting from prenatal life up to the adult animal. Cryo-cut sections of the three regions were incubated in the presence of the primary choline acetyltransferase (ChAT)-antibody and the immunoreaction was observed under an epifluorescence microscope and photographed. Our results demonstrate that cholinergic neurons are already present during prenatal life in the stomach and ileum, that several steps characterize cholinergic cell differentiation during postnatal life with a consistent delay in the appearance of ChAT-immunoreactivity (IR) in the submucous plexus compared to the myenteric plexus, and that the complete development is accomplished with weaning. In the colon the total number of ChAT-IR cells does not change from the suckling period to adulthood; a significantly larger number of ChAT-IR cells is found in the ileum of 5-day-old rats than in that of adult rats. PMID- 8710163 TI - Spontaneous periodic synchronized bursting during formation of mature patterns of connections in cortical cultures. AB - Long-term recording of spontaneous activity in cultured cortical neuronal networks was carried out using substrates containing multi-electrode arrays. Spontaneous uncorrelated firing appeared within the first 3 days and transformed progressively into synchronized bursting within a week. By 30 days from the establishment of the culture, the network exhibited a complicated non-periodic, synchronized activity pattern which showed no changes for more than 2 months and thus represented the mature state of the network. Pharmacological inhibition of activity only during the period when regular synchronized bursting was observed was capable of producing a different mature activity pattern from the control. These results suggest that periodic synchronized bursting plays a critical role in the development of synaptic connections. PMID- 8710164 TI - Molecular cloning and expression of the rat homologue of presenilin-1. AB - The rat homologue of the presenilin-1 (PS-1) gene, which is responsible for early onset familial Alzheimer's disease linked to chromosome 14, was cloned and sequenced. The predicted amino acid sequence showed quite high homology among rat, mouse, and human PS-1. Especially, the amino acid sequences of the putative transmembrane domains were highly conserved among the three species. The expression level of the PS-1 gene increased during brain development and the number of transcripts of the PS-1 gene changed during brain development. We found one transcript of the PS-1 gene in embryonic day 12 (E12)-E15 rat brain and two transcripts in E18-adult rat brain. Therefore, PS-1 may play a role in neurogenesis. PMID- 8710165 TI - The pyramidal cell layer of sector CA 1 shows the lowest hippocampal succinate dehydrogenase activity in normal and postischemic gerbils. AB - We examined regional differences in the activity of a mitochondrial respiratory enzyme, succinic dehydrogenase (SDH), in the hippocampi of normal and postischemic gerbils, using a quantitative imaging method. Gerbils (n = 21) without ischemia, and gerbils which had experienced 5 min of bilateral common carotid artery occlusion 12 h or 2 days previously, were sacrificed. Coronal sections of the brains were prepared for quantitative imaging of SDH activity and histological examination. In the control gerbils, SDH activity in the pyramidal cell layer of the CA 1 sector (Sommer's sector) was 106.3 +/- 10.3% (mean +/- SD; SDH activity as a percentage of the cerebellar SDH activity), which was lower than in the other subfields of the hippocampus. SDH activity in the oriens layer, stratum radiatum and lacunosum molecular layer of the CA 1 sector was lower than in the corresponding layers of the CA 2 and CA 3 sectors. After transient ischemia, SDH activity remained unchanged in the CA 1 sector. Histologically, selective neuronal necrosis was observed in the pyramidal cell layer of the CA 1 sector 2 days after ischemia. The observed low level of this mitochondrial respiratory enzyme in the pyramidal cell layer of the CA 1 sector should be taken into account as a possible trigger of the selective vulnerability of the region to ischemia. PMID- 8710166 TI - Cytosolic and membrane-bound cerebral nitric oxide synthase activity during hypoxia in cortical tissue of newborn piglets. AB - To determine the role of nitric oxide production during hypoxia, the presence of two forms of neuronal nitric oxide synthase, cytosolic (cNOS) and membrane-bound (memNOS), in cortical tissue of newborn piglets and the effects of hypoxia on the activity of these enzymes were studied. Experiments were performed in 12 anesthetized and ventilated Yorkshire piglets, 2-4 days of age. Hypoxia was induced by decreasing the FiO2 to 0.07. The control group was ventilated maintaining normoxia. After 1 h of normoxic or hypoxic ventilation brain tissue was removed and frozen immediately in liquid nitrogen. Tissue hypoxia was confirmed by analysis of adenosine triphosphate (ATP) and phosphocreatine (PCr): ATP was reduced to 52% and PCr to 28% of control values. cNOS activity was 35.3 +/- 13.7 pmol/mg protein per min in the control group and 28.3 +/- 7.0 in the hypoxia group; memNOS activity was 10.5 +/- 4.5 and 12.0 +/- 3.9 pmol/mg protein per min in the control and hypoxia groups, respectively. Differences in cNOS and memNOS activity between control and hypoxic animals were not significant. The results indicate that both cNOS and memNOS are present in cortical tissue of newborn piglets and that the activity is unaffected by 1 h of tissue hypoxia. We suggest that production of nitric oxide and its derivative peroxynitrite during hypoxia may therefore be a potential mechanism for hypoxia-induced brain cell membrane lipid peroxidation. PMID- 8710167 TI - Mental representations of morphologically complex words: an event-related potential study with adult humans. AB - Event related brain potentials (ERPs) were recorded from 13 subjects while performing a lexical decision task. Regular and irregular German principles were presented twice (identical repetition) or were primed by their related infinitives (morphological repetition). Regular participles primed by their infinitives displayed ERP waveforms similar to those evoked by identical repetitions, namely a large positivity with an onset latency of about 200 ms. Irregular participles, by contrast, showed no reliable morphological repetition effects. The results are discussed in terms of the controversy between approaches proposing a single representational mechanism for regular and irregular inflection and the "dual mechanism' account that suggests an associative memory for the storage of irregulars and a rule-based symbolic processor for generating regular forms. PMID- 8710168 TI - Intramembranous structure of the postsynaptic membrane in the rat sensorimotor cortex. AB - Intramembranous structure of the postsynaptic membrane of the axodendritic synapses in the rat sensorimotor cortex was examined by means of freeze-etching technique. Perforated and non-perforated aggregates of particles were found at the extracellular half (E-face) of the postsynaptic membrane. To study correlation between membrane structure and synaptic plasticity we compared size and particle packing density in both type aggregates of particles at the E-face of the postsynaptic dendritic spine membrane. The results are discussed in terms of plasticity on the synaptic contact zone (SCZ) at the postsynaptic membrane of the excitatory axospinous synapses. PMID- 8710169 TI - Distribution of visinin-like protein (VILIP) immunoreactivity in the hippocampus of the Mongolian gerbil (Meriones unguiculatus). AB - Visinin-like protein (VILIP) is a neuronal EF-hand Ca(2+)-binding protein. In the chick brain, it is widely expressed, e.g. in neurons of the visual pathway and the cerebellum. In the cerebellum, a presynaptic localization of VILIP in glutamatergic parallel- and climbing-fiber terminals has been observed. Here, we describe the distribution of immunoreactivity (IR) detected by antibodies against chick VILIP in the gerbil hippocampus at the light and electron microscopic level. VILIP antibodies stain neurons in the whole hippocampal formation including pyramidal cells in the CA1 and CA3 region of the Ammon's horn and granule cells of the dentate gyrus. In CA3 neurons, VILIP-IR is localized in dendrites and dendritic spines. PMID- 8710170 TI - Previous dopaminergic innervation is not necessary for the development of dopamine supersensitivity in rat striatal neurons. AB - The development of supersensitivity to dopamine by striatal neurons is widely considered to be a response to interruption of dopaminergic synaptic function. However, it is not clear whether pre-existing dopaminergic innervation is in fact necessary for supersensitivity to develop. In this study, rat foetal striatal cells were obtained prior to their innervation by dopaminergic fibres, and grafted into the cortex or thalamus of adult rats. In the absence of dopaminergic innervation, the grafts developed and became organized into striatum-like and non striatum-like patches. Both in animals with intrathalamic and in animals with intracortical grafts, systemic administration of a low dose of apomorphine (0.25 mg/kg) induced intense fos expression in striatum-like patches not innervated by dopaminergic fibres. These results indicate that pre-existing dopaminergic innervation is not required for the development of supersensitivity, and that dopaminergic innervation is necessary to develop an adequate synergistic interaction between D1 and D2 receptors. PMID- 8710171 TI - Ciliary neurotrophic factor attenuates spatial cognition impairment, cortical infarction and thalamic degeneration in spontaneously hypertensive rats with focal cerebral ischemia. AB - Ciliary neurotrophic factor (CNTF) has been shown to exhibit potent neurotrophic activity on peripheral and central neurons in vitro and in vivo. However, it remains to be determined whether or not CNTF rescues neuronal loss due to focal cerebral ischemia and prevents ischemia-induced disability of space navigation in rats. In the present in vivo study, we infused CNTF continuously for 4 weeks into the lateral ventricle, starting just after permanent occlusion of the left middle cerebral artery (MCA) of stroke-prone spontaneous hypertensive rats. CNTF infusion prevented the occurrence of ischemia-induced learning disability in a dose-dependent manner in rats subjected to the Morris water maze task. Subsequent histological examinations showed that cortical infarction and retrograde degeneration of the ipsilateral thalamic neurons in ischemic rats infused with CNTF were significantly less severe than those in ischemic rats infused with vehicle alone. These findings suggest that postischemic CNTF treatment prevents the occurrence of spatial learning disability in rats with permanent MCA occlusion, possibly by reducing neuronal damage within the cerebral cortex and secondary retrograde degeneration of the thalamus. PMID- 8710172 TI - Acute and chronic effects of estrogenic compounds on glutamate-stimulated phosphatidylinositol metabolism in primary neuronal cultures. AB - Glutamate (Glu)-stimulated phosphatidylinositol (PI) metabolism in primary neuronal cultures was found to be modulated by acute and chronic treatment with two estrogenic compounds. 17 beta-Estradiol 3-benzoate (0.1 and 1 microM), when applied with Glu, significantly reduced Glu (40 microM)-stimulated PI metabolism by 20-36%, an effect not seen with 17 alpha-estradiol. The weak estrogen phenol red (20 microM), had no effect when added immediately before Glu stimulations. Two-week pretreatment with 17 beta-estradiol 3-benzoate (1 microM) resulted in a significant decrease in Glu-stimulated PI metabolism (10-100 microM). Chronic treatment with 20 microM phenol red, at a concentration commonly found in culture medium, resulted in parallel but not statistically significant effects to those observed with chronic estradiol treatment. Estrogenic compounds may modulate the excitatory responses of neurons by both genomic and non-genomic means. PMID- 8710173 TI - Early increases in TNF-alpha, IL-6 and IL-1 beta levels following transient cerebral ischemia in gerbil brain. AB - The effects of transient global ischemia using bilateral carotid artery occlusion on regional cytokine levels in gerbil brain were investigated using enzyme-linked immunoassay techniques. Brain concentrations of interleukin-6 (IL-6), interleukin 1 beta (IL-1 beta), and tumor necrosis factor-alpha (TNF-alpha) were increased during the early recirculation period ( < 6 h) after 10 min of ischemia, with lesser degrees of elevation following only 5 min of ischemia. TNF-alpha levels in the hippocampus and striatum were significantly increased as early as 1 h after recirculation, declining sharply to control levels by 12 h, then transiently increasing at 24 h. Elevated levels of IL-1 beta and IL-6 were not seen until 3-6 h post-occlusion. No significant increases in cytokine concentrations were observed in the cerebellum or thalamus. These results suggest that regionally selective increases in cytokines may be involved in the pathophysiological changes in hippocampus and striatum following transient cerebral ischemia. PMID- 8710174 TI - Cyclic GMP inhibits phosphoinositide turnover in choroid plexus: evidence for interactions between second messengers concurrently triggered by 5-HT2C receptors. AB - The present study examined the effects of the nitric oxide generator sodium nitroprusside (SNP), a membrane-permeable cGMP analog (dibutyryl-cGMP) and low calcium buffer incubation on choroid plexus serotonin 5-HT2C receptor-mediated inositol monophosphate (IP) production. SNP (100 microM) substantially inhibited 10(-6)M serotonin-stimulated IP production (-46%, P < 0.02). Serotonin-stimulated IP production was increased in low calcium buffer (+280%, P < 0.01) in which serotonin-stimulated cGMP formation is attenuated. Addition of dibutyryl-cGMP (500 microM) inhibited IP formation in low calcium buffer. The present data are suggestive of an inhibitory effect of cGMP on IP formation in choroid plexus, and raise the intriguing possibility of interactions between second messenger systems concurrently activated by 5-HT2C receptors. PMID- 8710175 TI - Blood-brain barrier uptake of the 40 and 42 amino acid sequences of circulating Alzheimer's amyloid beta in guinea pigs. AB - An intracarotid brain infusion/capillary depletion technique was used in guinea pigs to examine cerebral capillary sequestration and transport into brain parenchyma of sA beta 1-40 and sA beta 1-42, synthetic peptides identical to two forms of the amyloid beta peptide found in Alzheimer's disease lesions: the 40 residue form, found primarily in vascular deposits, and the 42 residue form, found primarily in senile plaques. The peptides crossed well into the brain parenchyma via a specific transport mechanism for which sA beta 1-40 had an approximately two-fold greater affinity than sA beta 1-42. There was significant capillary sequestration of sA beta 1-40, but retention by the microvasculature of sA beta 1-42 was negligible. These data suggest that the level of the 40 residue peptide in cerebral vasculature and of the 42 residue peptide in parenchyma could be regulated by blood-brain barrier sequestration and transport of their respective circulating precursors. PMID- 8710176 TI - Evidence for colocalization of GABA and glycine in afferents to retrogradely labelled rat abducens motoneurones. AB - The coexistence of gamma-aminobutyric acid (GABA) and glycine in axon terminals impinging on rat abducens motoneurones was investigated using a double staining procedure combining retrograde labelling of the motoneurones with HRP and post embedding immunocytochemical staining of axon terminals. Adjacent ultrathin sections of cell bodies of identified motoneurones were individually treated with GABA or glycine antibodies. The terminals single labelled for GABA represented 11.4% of the terminals analyzed, while 8% of them were glycine immunoreactive and 9% were both GABA and glycine immunoreactive. All the labelled terminals contained pleomorphic vesicles. The mean length of apposition of the double labelled terminals was statistically larger (2.20 +/- 0.97 microns) than the GABA (1.65 +/- 0.57 microns) or glycine immunoreactive ones (1.37 +/- 0.35 microns). PMID- 8710177 TI - Alterations in urinary bladder synaptosomal neurotransmitter concentrations in two-week streptozotocin-induced diabetic rats. AB - Three-month-old male Wistar rats were rendered diabetic with a single intravenous injection of streptozotocin (60 mg/kg body weight). Two weeks after induction of diabetes, synaptosome-rich fractions were prepared from urinary bladder tissue homogenate of the diabetic rats and control rats by differential centrifugation (1000 x g, 17,000 x g and 100,000 x g) with discontinuous sucrose gradient. Synaptosomal acetylcholine, norepinephrine, epinephrine and dopamine were measured by the method of high-performance liquid chromatography. The respective neurotransmitter concentrations for the diabetic rats were 1537.8 +/- 65.3, 4757.7 +/- 361.9, 3720.7 +/- 276.1, and 2447.8 +/- 196.8 pmol/mg synaptosomal protein, respectively; those for the control rats were 338.1 +/- 25.0, 1009.0 +/- 54.6, 645.3 +/- 52.2, and 1426.1 +/- 123.9 pmol/mg protein, respectively. Thus, the synaptosomal concentrations for all the measured neurotransmitters were significantly higher in the diabetic rats (P < 0.05 for each comparison). In conclusion, it has been demonstrated that the vesicle-bound acetylcholine and catecholamines in the synaptosome-rich fraction of the urinary bladder were significantly increased in 2-week diabetic rats. This finding would suggest impaired neurotransmitter release from both the bladder sympathetic and parasympathetic efferent nerve endings in early streptozotocin-induced diabetes. PMID- 8710178 TI - Granules in glial cells of patients with Alzheimer's disease are immunopositive for C-terminal sequences of beta-amyloid protein. AB - Granular structures that are recognized by antibodies specific for the C-terminal but not the N-terminal sequences of the beta-amyloid protein (A beta) fragments are present in a subset of microglia and astrocytes in Alzheimer brain tissue. The immunohistochemical profile indicates that the A beta in these granules is truncated between the residues 17 and 31 and terminates at the residue 42 or 43. Such granule-containing glia occur only in brain areas with the heavy A beta deposits. Whether the intraglial A beta fragments accumulate as a result of phagocytosis of extracellular A beta or are formed intracellularly by glial cells from amyloid precursor protein (APP) remains unknown. PMID- 8710179 TI - Molecular forms of acetyl- and butyrylcholinesterase in human glioma. AB - Specimens of astrocytoma, oligodendroglioma and medulloblastoma were sequentially extracted with saline and saline-Triton X-100 buffers. Acetyl- (AChE) and butyrylcholinesterase (BuChE) activities were assayed in the soluble fractions, these being further analyzed to establish the distribution of molecular forms. All the tumors tested showed AChE and BuChE activities, the measured AChE/BuChE ratios being unrelated to the malignant grading. Hydrophilic and amphiphilic AChE and BuChE tetramers, amphiphilic AChE dimers and monomers, and hydrophilic BuChE monomers were identified in all the tumors analyzed. The amphiphilic behavior of the enzyme forms was assessed by sedimentation analysis and hydrophobic chromatography on phenyl-Agarose. A small fraction of glioma AChE monomers was released as, or transformed into, hydrophilic forms by incubation with phosphatidylinositol-specific phospholipase C (PIPLC). These data suggest that AChE monomers bearing distinct hydrophobic domains coexist in human glioma. PMID- 8710180 TI - Effect of rapid eye movement sleep deprivation on 5'-nucleotidase activity in the rat brain. AB - Adenosine has been implicated in the regulation of rapid eye movement sleep (REMS). In an attempt to understand the mechanism of production of adenosine in relation to REMS it was hypothesized that should it be involved in REMS, the latter's deprivation is likely to affect its synthetic machinery. Hence, male albino rats were deprived of REMS by the flower pot technique and the activity of 5'-nucleotidase, an enzyme responsible for adenosine synthesis, was estimated in the cerebrum, cerebellum and brain stem. Suitable control experiments were conducted to rule out the non-specific effects. The results showed that 5' nucleotidase activity decreased only after 4 days deprivation and in the cerebrum only; while short-term (2 days) deprivation did not affect the enzyme activity in any of the brain areas. The altered enzyme activity returned to baseline level after recovery from REMS deprivation. The results from other control experiments suggested that the effects were primarily due to REMS deprivation and not due to non-specific factors. It is proposed that if adenosine is involved in REMS, its production is unlikely to depend on 5'-nucleotidase or it may account primarily for EEG desynchronization. PMID- 8710181 TI - Cyclic AMP and pituitary adenylate cyclase-activating polypeptide (PACAP) prevent programmed cell death of cultured rat cerebellar granule cells. AB - Cultured cerebellar granule cells undergo programmed cell death when they are deprived of depolarizing KCl. Results from this study indicate that the cAMP analog cyclic 8-(4-chlorophenylthio) adenosine-3'5'-monophosphate (CPT-cAMP) can prevent the cell death in a dose-dependent manner, with the maximal effect seen at 500 microM. Approximately 70% of cells can be saved with this concentration of CPT-cAMP for at least 6 days. Pituitary adenylate cyclase-activating polypeptide (PACAP) increased the intracellular cAMP levels of these cells in a time- and dose-dependent manner. This agent can prevent the decrease of cAMP and cell death induced by KCl withdrawal. These results suggest that PACAP could function as a neurotrophic factor for cerebellar granule cells in vivo. PMID- 8710182 TI - Histochemical evidence of the increased expression of nicotinamide adenine dinucleotide phosphate-dependent diaphorase in neurons of the myenteric plexus after acute spinal cord injury in adult rats. AB - The expression of nitric oxide synthase in neurons of the gastrointestinal tract (GIT) after experimental spinal cord injury (SCI) was assessed in adult rats contused at T8. One day and 10 weeks after injury, specimens along the GIT were studied for NADPH-diaphorase histochemistry. A significant increase in the number of positive cell bodies and fibers in the myenteric plexus were observed 1 day after SCI, as compared to specimens from control and chronically injured rats, with the exception of the colon, which showed unchanged or decreased number of positive neurons in the acute and chronic stages, respectively. Positive neurons in the submucous plexus remained unchanged, excepting an increase in the colon after acute SCI, and a decrease in the duodenum in chronically injured rats. The altered nitric oxide neurotransmission in the GIT may be relevant to its reduced motility after SCI. PMID- 8710183 TI - Dephosphorylation studies of SKNSH-SY 5Y cell Tau proteins by endogenous phosphatase activity. AB - Recent data have shown that the microtubule-associated Tau proteins are phosphorylated but to a lesser extent than PHF-Tau proteins which are the major components of Alzheimer's disease paired helical filaments. These normal Tau proteins are highly sensitive to the endogenous phosphatase activity during post mortem delay. In order to understand the basic equilibrium between phosphatase and kinase activities, phosphorylation and dephosphorylation mechanisms of Tau proteins were studied in neuroblastoma cells. The present results demonstrate that an endogenous phosphatase activity is present and directed on Tau proteins in the SKNSH-SY 5Y cell extracts. Interestingly, the okadaic acid-induced hyperphosphorylated Tau proteins are more resistant to the phosphatase activity than the control Tau proteins. Our data emphasize the value of this in vitro cellular model for the study of biological conditions that control Tau protein phosphorylation levels. PMID- 8710184 TI - Polyamines in frontal cortex of patients with Down syndrome and Alzheimer disease. AB - Polyamines may play an important role in brain development, mature brain function and also in neurodegenerative conditions. We investigated polyamine levels in frontal cortex of human post-mortem brain samples of elderly patients with Down syndrome (DS), Alzheimer disease (AD) and normal controls by means of chromatographic separation after dansylation. Spermidine and spermine concentrations were markedly decreased in DS and AD. Polyamine levels were neither related to age and post-mortem interval nor to choline acetyltransferase activity, as indicator of neuronal loss. Our results support the idea that besides other neurotransmitter systems, endogenous polyamine levels are altered in dementing illnesses such as Alzheimer disease and Down syndrome. PMID- 8710185 TI - No evidence of linkage between schizophrenia and D2 dopamine receptor gene locus in Italian pedigrees. AB - Our purpose was to test the dopamine D2 receptor gene (DRD2), the tyrosine hydroxylase (TH) gene and the monoamino oxydase A (MAO-A) gene for linkage to schizophrenia and bipolar disorders. We have analyzed seven Italian families with schizophrenia and four families with bipolar disorders for a total of 68 individuals; 32 individuals were affected. Diagnoses were made using the structured clinical interview Schedule for Affective Disorders and Schizophrenia, Lifetime version (SADS-L). The results of our study provide no evidence of linkage between alleles at D2 dopamine receptor loci and schizophrenia or bipolar disorders. The markers TH gene and MAO-A gene give slightly positive or negative results suggesting the utility of further analysis on more informative families. PMID- 8710186 TI - Effect of hypoxia and reoxygenation on regional activity of nitric oxide synthase in brain of newborn piglets. AB - The activity of nitric oxide synthase (NOS) was measured in homogenates from cortex, striatum, hippocampus, cerebellum, pons, thalamus and midbrain of the brain of newborn piglets and the effects of hypoxia and posthypoxic period on this activity was evaluated. The control activities were 19.7, 31.5, 26.8, 16.7, 33.6, 19.3 and 39.4 pmol/mg protein per min, respectively. A 1 h period of hypoxia (an FiO2 of 7%) resulted in statistically significant decreases in the activity of NOS in every region of the brain except for the cortex, where the activity was not significantly altered compared to control. By 2 h of reoxygenation following such a hypoxic episode, the NOS activities increased to above control levels in all regions of the brain, but this increase was statistically significant compared to control only in thalamus. Since hypoxia induced the greatest decrease in NOS activity in the cerebellum, the kinetic constants of the enzyme were measured in homogenates from this region of brain. The decreased activity following the hypoxic episode was associated with an approximately four-fold increase in the apparent affinity (KM) for arginine with no significant change in the maximal rate of reaction (Vmax). The decrease in NOS activity subsequent to a hypoxic episode may contribute to the disturbances in cellular metabolism in the immature brain induced by episodes of hypoxia reoxygenation. PMID- 8710187 TI - Magnetometry of evoked fields from human peripheral nerve, brachial plexus and primary somatosensory cortex using a liquid nitrogen cooled superconducting quantum interference device. AB - Superconducting Quantum Interference Devices (SQUIDs) can be used to detect neuromagnetic fields evoked in the peripheral and central nervous system. Up to now, such measurements had to be based on SQUIDs with a low critical temperature (Tc) requiring liquid helium cooling. Recent improvements in high-Tc SQUID technology relying on liquid nitrogen cooling led to a significant reduction in the system's noise level. Hare, first high-Tc recordings of weak neuromagnetic fields are demonstrated. In particular, along the entire somatosensory afferent pathway including peripheral nerves, brachial plexus and primary somatosensory neocortex evoked neuromagnetic activities were detected using conventional recording parameters for bandwidth and number of averages. This opens up a wide perspective for cost-effective high-Tc magnetometry in clinical neuroscience. PMID- 8710188 TI - Regulation of glucocorticosteroid receptor expression in rat hippocampal cell cultures by nerve growth factor. AB - Dispersed hippocampal cells cultured in serum-free conditions were used to study the effects of nerve growth factor (NGF) on the expression of type I (mineralocorticosteroid or MR) and type II (glucocorticosteroid or GR) corticosteroid receptors. Cells, plated at a density of 1.2 x 10(6) cells/ml in 60 mm Petri dishes, were mainly identified as neurons (90-95%) and maintained for at least 2 weeks. A 7-day treatment with 10-50 ng NGF/ml induced a concentration dependent decrease of GR binding (40% decrease) compared to untreated cells. In contrast, MR density was unaffected by a 7-day treatment with 50 ng NGF/ml. Data are discussed as possible direct and/or indirect effects of NGF at the level of both neuronal and glial cells. PMID- 8710189 TI - Derangement in stress response of apolipoprotein E-deficient mice. AB - Apolipoprotein E (apoE) is associated with familial and sporadic Alzheimer's disease (AD). Stress has been identified as a putative risk factor of AD. Thus, in the present study we examined the susceptibility of apoE-deficient mice to stress. The results obtained revealed that the elevation of corticosterone levels in apoE-deficient mice following restraint stress is markedly lower than in controls, and that these mice differ in their behavioral pain response to noxious stimuli in both stress and non-stress conditions. These findings suggest an interplay between apoE and the response to stressful stimuli and provide a model for elucidating the relationship between apoE and susceptibility to stress. PMID- 8710190 TI - Chronic ethanol consumption increases the amount of mRNA for retinoic acid and triiodothyronine receptors in mouse brain. AB - It is known that alcohol induces disorders in the metabolism of retinoids and particularly in the biosynthetic pathways of retinoic acid (RA). Since RA has, along with other hormones and particularly triiodothyronine (T3), a physiological role in the adult brain, the effect of chronic exposure to alcohol on RA and T3 status was investigated. The amounts of RA receptor (RAR) and T3 receptor (TR) mRNAs were quantified and the activity of the 'tissue' transglutaminase (tTG; an RA-dependent enzyme) was assayed in the brain of mice following chronic ethanol consumption (CEC; 12% v/v for 6-10 months). Compared to controls, ethanol-treated mice exhibited increased amounts of RAR and TR mRNAs together with an increase in tTG activity. It is hypothesized that the enhanced cellular action of RA and T3 could play a role in the previously described brain damages induced by CEC. PMID- 8710191 TI - Is the center of gravity controlled during upper trunk movements? AB - The question was addressed in this study as to whether the kinematic synergy responsible for equilibrium control during upper trunk movements may involve an actual evaluation of the weight of the body segments. Five adult subjects were asked to bend the upper trunk forward or backward to an angle of about 35 degrees, first without any load and then with a 10 kg load fixed to their shoulders. The center of gravity (CG) shift in the sagittal direction which occurred at the end of the movement was compared versus without a load. Two patterns of CG shift were identified. In the first pattern shown by three subjects in forward and three in backward trunk bending, the CG shift remained the same, while in the other pattern (two subjects for forward and two for backward trunk bending), the CG shift increased by an amount corresponding to the purely mechanical effect of the load. These results indicate that the actual weight of the segments may be evaluated on the basis of sensory inputs and may be responsible for a change in the kinematic synergy which preserves the CG control during the upper trunk movement. PMID- 8710192 TI - Neuroprotective effect of FK506, an immunosuppressant, on transient global ischemia in gerbil. AB - Delayed neuronal death (DND) in CA1 region after transient global ischemia is a well-known phenomenon, but its mechanism has not been clarified. In order to examine the involvement of nitric oxide (NO) in DND, 7-nitro indazole (7NI), a selective neuronal NO synthase (nNOS) inhibitor, and FK506, an immunosuppressant which also inhibits nNOS, were administered intraperitoneally during and after transient global ischemia in gerbil. FK506 moderately ameliorated DND in a dose dependent manner. However, 7NI showed only minor neuroprotective effects. These results show that DND is not mainly mediated by NO production via nNOS, and FK506 acts as a neuroprotective agent via unknown pathways other than nNOS inhibition. PMID- 8710193 TI - Abnormal, ubiquitinated cortical neurites in patients with diffuse Lewy body disease. AB - Although it is known that the severity of dementia in patients with diffuse Lewy body disease is related to cortical Lewy body density, the morphological substrate of dementia in these patients is poorly understood. Vibratome sections processed free-floating for ubiquitin immunohistochemistry in three patients with the common form and one patient with the pure form has shown the presence of large numbers of abnormal, ubiquitinated neurites in the cerebral cortex, mainly cingulate cortex, entorhinal cortex and temporal lobe, and hippocampal complex, regions in which, precisely, Lewy bodies are most abundant. Abnormal neurites are a consistent change which results in abnormal neuronal connectivity. Abnormal cortical neurites, rather than cortical Lewy bodies, may play a significant role in the development and progression of cognitive deficits in patients with diffuse Lewy body disease. PMID- 8710194 TI - Different effect of height on latency of leg and foot short- and medium- latency EMG responses to perturbation of stance in humans. AB - In standing humans, platform perturbations evoked short- (SLR) and medium-latency responses (MLR) in soleus (Sol), tibialis anterior (TA) and flexor digitorum brevis (FDB) muscles. The latency of all responses significantly increased with subjects' height. The slope of the regression lines for the MLRs versus height was significantly steeper than that for the SLRs. The conduction velocity of the afferent fibres mediating the FDB MLR, calculated on the basis of the regression, was 17.5 m/s. These findings indicate that the MLRs are transmitted through afferent fibres slower than those mediating the SLRs. While the latter fibres are spindle group Ia afferents, the former are suggested to be group II fibres. PMID- 8710195 TI - Effects of different patterns of light adaptation on cellular and synaptic plasticity in teleost retina: comparison of flickering and steady lights. AB - The importance of the pattern of light stimulus in inducing light-adaptive morphological (cellular and sub-cellular) changes in the outer retina of the carp was assessed. Thus, the effects of steady and flickering backgrounds (of the same intensity) on cone photomechanical movements (PMMs) and horizontal cell (HC) spinules were compared by quantitative measurements. For both phenomena, flickering stimuli had a significantly greater effect than steady, although the number of photons delivered by the former was one-half. The results demonstrate clearly that the pattern of light stimulus is indeed an important parameter in determining the extent of adaptation in the outer retina. The effects are discussed in relation to possible neurochemical bases of light-adaptive neural control mechanisms. PMID- 8710196 TI - Nerve growth factor phase shifts circadian activity rhythms in Syrian hamsters. AB - Nerve growth factor (NGF) receptors are found in high density in the rodent suprachiasmatic nucleus (SCN), a site which regulates mammalian circadian rhythms. We examined the effects of NGF (40 ng) or vehicle injections into the SCN, at circadian times (CT) 6, 14 or 22 on activity rhythms in hamsters maintained in constant darkness. NGF caused phase advances at CT6 (30.9 min) and CT22 (36.9 min), and phase delays at CT14 (31.2 min). Saline and cytochrome-c administration had no phase-shifting effects at CT6 and CT22, but at CT14 cytochrome-c produced large phase delays, implying that NGF-induced delays at this phase may be non-specific. Similarities between NGF-induced shifts and those elicited by the cholinergic agonist carbachol suggest a common mode of action. PMID- 8710197 TI - Increasing gonadotropin-releasing hormone release by perifused hypothalamus from early to late anestrus in the beagle bitch. AB - An in vitro perifusion system was used to investigate pulsatile gonadotropin releasing hormone (GnRH) release from hypothalamic fragments derived from beagle bitches at different stages of the estrous cycle. The spontaneous GnRH release from the excised tissue fragments that include the "mediobasal hypothalamic preoptic area-suprachiasmatic nucleus units' was episodic throughout all stages of the estrous cycle with a significantly high release rate during late anestrus and late proestrus. The GnRH release rate and plasma levels of luteinizing hormone were positively correlated (r = 0.94, P < 0.01). These results suggest that during the course of anestrus in the bitch the GnRH release rate increases while the pituitary responds accordingly. PMID- 8710198 TI - Regional and cellular distribution of spermidine/spermine N1-acetyltransferase (SSAT) mRNA in the rat central nervous system. AB - Spermidine/spermine N1-acetyltransferase (SSAT) is the key enzyme responsible for polyamine interconversion. SSAT mRNA (visualized by in situ hybridization histochemistry) was shown to have a wide but heterogeneous distribution in the central nervous system (CNS) at both regional and cellular levels. The highest labelling was observed in hippocampus (pyramidal and polymorph neurons) and olfactory bulb. Present data suggest that polyamine metabolism in the CNS is not homogeneous but rather that the preferential production of a polyamine species is region- and cell type-specific. PMID- 8710199 TI - Delivery of membrane-impermeant fluorescent probes into living neural cell populations by lipotransfer. AB - Use of fluorescent probes to monitor f-actin in living cells typically relies on difficult microinjection procedures. The current work has developed cationic lipotransfer of membrane-impermeant probes as an alternative to microinjection. BODIPY FL-phallacidin, a fluorescent f-actin probe, was packaged into 40-50 nm cationic liposomes. Packaging, verified by gel filtration, enabled delivery of the probe into living nerve cells and provided an image of f-actin that was identical to that seen in fixed, permeabilized cells. Phallacidin alone did not enter living cells, nor was its uptake stimulated by the presence of empty liposomes. All predicted f-actin structures were fluorescent in living cells, indicating a high efficacy of delivery. Cationic lipotransfer of fluorescent probes was rapid, not disruptive to cells, and delivered a probe en masse to a large sample population. Lipotransfer appears to be a promising alternative to microinjection for introducing membrane-impermeant probes and reagents into living cells. PMID- 8710200 TI - Reinnervation after destruction of the dopaminergic system in the rat nucleus accumbens: a quantitative immunohistochemical analysis. AB - The recurrence of dopamine-immunoreactive (DAi) fibers and the effect of the adrenocorticotrophic hormone (ACTH)-(4-9) analog ORG 2766 on this process were investigated 1, 4, 12 and 24 weeks after a unilateral 6-hydroxydopamine (6-OHDA) lesion in the nucleus accumbens (NAc). DAi fibers were almost completely absent 1 week after the lesion. A gradual increase in DAi fibers throughout the NAc was observed, with subnormal values at 24 weeks. Treatment with ORG 2766 during the first week after the lesion resulted in more DAi fibers 4 weeks after the lesion as compared to placebo treatment, but not 12 and 24 weeks after the lesion. After 6-OHDA lesioning reinnervation of the NAc takes place and this process is transiently facilitated by ORG 2766. PMID- 8710201 TI - Neuromodulation of frontal and temporal cortex by intravenous d-fenfluramine: an [15O]H2O PET study in humans. AB - This study assessed the modulatory effect of a serotonergic agonist, d fenfluramine, on localized neuronal firing as indexed by changes in regional cerebral blood flow (rCBF). Previously, we reported the effect of oral d, l fenfluramine on neuronal activity as measured by change in [18F]fluorodeoxyglucose uptake. Improvements in the current study include: a more specific serotonin agonist, d-fenfluramine; a more reliable administration route, intravenous; and a one session paradigm made possible with the radiotracer [15O]H2O. Changes in relative rCBF (P < 0.001) were observed: increases within the frontal cortex bilaterally and decreases within the temporal cortex bilaterally, and left thalamus. Other significant findings were elevated cortisol and growth hormone; increased euphoria and panic symptoms and decreased tiredness. These results support further investigation with intravenous d fenfluramine to study the net functional effects of serotonergic stimulation in health and illness. PMID- 8710202 TI - Bilaterally enhanced dorsal horn postsynaptic currents in a rat model of peripheral mononeuropathy. AB - Surface compound potentials were recorded from the surgically exposed lumbar spinal cord in anaesthetized rats which had had one sciatic nerve loosely ligatured 12-15 days previously, resulting in unilateral allodynia and hyperalgesia, as assessed behaviourally. These cord dorsum potentials were recorded in response to electrical stimulation of the ligatured and non-ligatured sciatic nerve, respectively, on both the ipsi- and contralateral side with respect to the stimulated nerve. Compared to potentials produced by stimulation of the non-ligatured sciatic nerve, electrical stimulation of large diameter fibres proximal to the ligatures resulted in a smaller afferent fibre input arriving at the spinal cord. However, larger net postsynaptic currents in the contralateral dorsal horn and a larger net postsynaptic current per unit of afferent fibre input were found in the ipsilateral and contralateral spinal cord. Such changes may result from structural changes or increased synaptic efficacy in the dorsal horn following peripheral nerve injury. PMID- 8710203 TI - Effect of spinal infusion of L-NAME, a nitric oxide synthase inhibitor, on spinal tolerance and dependence induced by chronic intrathecal morphine in the rat. AB - Systemic L-NG-nitroarginine methyl ester or L-NAME (LN), a nitric oxide synthase inhibitor, has been reported to attenuate systemic opioid tolerance and withdrawal. Intrathecal co-administration of LN (100 and 500 nmol/microliter per h) with spinal morphine produced only a small diminishing attenuation of tolerance and attenuated only one of seven signs of withdrawal. These results show that LN has little effect on morphine tolerance and withdrawal at spinal sites. PMID- 8710204 TI - Diazepam increases melatonin secretion of photosensitive pineal organs of trout in the photopic and mesopic range of illumination. AB - The pineal organ of teleost fish receives photic information directly through specialized photoreceptor cells that transmit their light response to second order neurons and respond also with an endocrine light-dependent melatonin signal. In the present study we have analyzed the action of diazepam, a full agonist of the benzodiazepine receptor, on the photic regulation of the endocrine melatonin response of cultured trout pineal organs. Melatonin release of explanted pineal organs was clearly dependent on the irradiance of incident light with a maximum change during mesopic illuminations. Addition of diazepam to the superfusion medium significantly increased melatonin production in the mesopic and partly in the photopic range of illumination, without showing clear effects in the dark-adapted organ. Flumazenil, a central acting benzodiazepine antagonist, slightly reduced melatonin secretion. The action of diazepam appears to be comparable to a dark-pulse in the mesopic range of illuminations. PMID- 8710205 TI - Differential sensitivity to Mg(2+)-and tubocurarine-block of frog neuromuscular junctions in summer and winter. AB - Several parameters of transmitter release in neuromuscular junctions were compared in "winter' and "summer' frogs. In low Ca2+/high Mg(2+)-block, the quantal content m of endplate potentials was similar for both groups. In curarized junctions, however, endplate potentials were about twice as large in winter as in summer, and facilitation was lower and depression higher. This indicates that transmitter release is higher in winter junctions. This is not reflected in Mg(2+)-block, suggesting that strong calcium deprivation may suppress release in winter more strongly than in summer junctions. PMID- 8710206 TI - Changes in spontaneous unit activity in lumbar spinal cord after reversible aortic occlusion in the rat. AB - Lumbar dorsal horn neuronal multiunit activity was recorded in the halothane anesthetized rat before, during, and after a 30 min interval of spinal cord ischemia induced by inflation of a balloon catheter inserted through the femoral artery. After initiation of ischemia, there was typically a brief burst of neuronal activity, followed by electrical silence. During reperfusion, there was a progressive increase in discharge activity over the ensuing 30 min. Spontaneous activity was significantly increased, as measured by the number of discriminable cells and frequency of discharges. Post-ischemic hyperactivity reached maximal values typically after 60-90 min of reperfusion. The character and time course of recorded activity are consistent with the behavioral sequelae of transient spinal ischemia. PMID- 8710207 TI - Head-activator and the neuroectodermal differentiation of P19 mouse embryonal carcinoma cells. AB - P19 embryonal carcinoma cells differentiate into neuroectodermal derivates upon aggregation and treatment with retinoic acid (RA). Such RA-induced P19 cells, concomitant with neuronal differentiation, produce the neuropeptide head activator (HA) and release HA into the culture medium. The amount of HA produced by such differentiating P19 cells is more than 100-fold higher than that of uninduced P19 cells. RA-induced P19 cells respond to HA treatment with a two-fold increase in total number of neuronally differentiating P19 progeny. As early response to HA, an increase in intracellular calcium concentration is observed. PMID- 8710208 TI - alpha-Tocopherol quinone level is remarkably low in the cerebrospinal fluid of patients with sporadic amyotrophic lateral sclerosis. AB - In order to investigate the role of free radicals in the pathogenesis of sporadic amyotrophic lateral sclerosis (SALS), the concentrations of alpha-tocopherol (alpha-TOH) and its oxidized form alpha-tocopherol quinone (alpha-TQ) in the cerebrospinal fluid (CSF) of SALS patients were determined. The alpha-TOH level was 31% lower (P < 0.05) and the alpha-TQ level was 75% lower (P < 0.001) in SALS patients than in normal subjects. The results of the present study do not support the hypothesis that activated lipid peroxidation accelerates oxidation of alpha TOH into alpha-TQ in SALS patients. PMID- 8710209 TI - The origins of catecholaminergic innervation in the rostral ventromedial medulla oblongata of the rat. AB - The localization of catecholaminergic neuronal cell bodies which project to the rostral ventromedial medulla oblongata (RVM) were investigated by the combined technique with dopamine beta-hydroxylase (DBH), phenylethanolamine N methyltransferase (PNMT) or tyrosine hydroxylase (TH) immunocytochemistry and retrograde neuronal tracing method using fluorescent latex microspheres (FluoSpheres) injected into the center of the RVM, the nucleus raphe magnus (NRM). Noradrenaline (NA) neurons in A1, A5, A7 regions, locus coeruleus (LC) and nucleus subcoeruleus (SC) and adrenaline (Ad) neurons in C1 region were double labeled due to DBH or PNMT and retrogradely transported FluoSpheres, and the ratio of their coexistence was higher in A1, A5, A7 and C1 than in LC and SC. No dopamine neurons in the midbrain and forebrain were double-labeled with TH and FluoSpheres. Thus, it was clarified that the RVM is innervated by the ventral groups of lateral tegmental NA and Ad neurons in the brainstem. PMID- 8710210 TI - Stable functional expression of the adult subtype of human muscle acetylcholine receptor following transfection of the human rhabdomyosarcoma cell line TE671 with cDNA encoding the epsilon subunit. AB - The human rhabdomyosarcoma cell line TE671 expresses the foetal subtype of muscle acetylcholine receptor (AChR). By transfecting TE671 cells with cDNA encoding the human muscle AChR epsilon subunit under the control of the cytomegalovirus promoter we have established a stable cell clone that, in addition, constitutively expresses the adult AChR subtype. Both subtypes are inserted into the plasma membrane and demonstrate their respective characteristic single channel properties. The level of expression of the adult AChR subtype is two- to three-fold higher than that of the foetal subtype. The new cell clone provides a relatively abundant source of human adult AChR for immunological and pharmacological investigations. PMID- 8710211 TI - Presynaptic localization of a metabotropic glutamate receptor, mGluR8, in the rhinencephalic areas: a light and electron microscope study in the rat. AB - The present study indicated presynaptic localization of a metabotropic glutamate receptor, mGluR8, in projection neurons of the main olfactory bulb of rat. An antibody was produced by using a peptide corresponding to C-terminal 23 amino acids of mouse mGluR8. It was confirmed that the C-terminal 23 amino acids of rat mGluR8 were the same as those of mouse mGluR8 except for one, and that the antibody specifically recognized mGluR8 in the rat rhinencephalon. In layer Ia of the piriform cortex (a target area of projection fibers from the main olfactory bulb), mGluR8-like immunoreactivity (mGluR8-LI) was reduced after transection of the lateral olfactory tract, and mGluR8-LI was observed in axon terminals which were filled with round synaptic vesicles and made asymmetric synapses with dendritic spines. PMID- 8710212 TI - In situ hybridization study of myelin protein mRNA in rats with an experimental diabetic neuropathy. AB - Distribution of protein zero (P0) and myelin basic protein (MBP) mRNAs in the sciatic nerve from rats with alloxan-induced diabetes was analyzed at two different time points using in situ hybridization. Some animals of each diabetic group were treated with insulin. Densitometric quantitation of silver clusters revealed that 5 weeks after diabetes induction P0 mRNA only is significantly increased, while at 14 weeks both P0 and MBP mRNA contents are markedly higher than controls. Insulin treatment normalizes glycemia levels and slightly counteracts increased P0 mRNA at both stages of diabetes. An increase in MBP mRNA is observed in chronic diabetic animals only, and is unaltered by the normoglycemic effect of insulin. The increased transcript levels of P0 and MBP suggest that Schwann cells can modulate gene expression of myelin-specific proteins in response to diabetic-induced metabolic derangement. Such a change may represent a higher turnover of myelin proteins as an attempt by the Schwann cells to repair the diabetes-induced nerve damage. The observed pattern of transcript amount is only slightly influenced by insulin treatment. PMID- 8710213 TI - A metabotropic glutamate receptor agonist DCG-IV suppresses synaptic transmission at mossy fiber pathway of the guinea pig hippocampus. AB - The effects of specific metabotropic glutamate receptor (mGluR) agonists on field excitatory postsynaptic potentials (fEPSPs) at mossy fiber-CA3 synapses were examined in guinea pig hippocampal slice preparations. Application of a novel and potent group II-selective mGluR agonist (2S,1'R,2'R,3'R)-2-(2,3 dicarboxycyclopropyl)glycine (DCG-IV; 0.1 microM) reversibly reduced the fEPSPs. Both the group III-selective agonist DL-2-amino-4-phosphonobutyric acid (AP4; 50 microM) and the broad-spectrum agonist 1S,3R-1-aminocyclopentane-1, 3 dicarboxylic acid (1S,3R-ACPD; 5 microM) also reversibly suppressed the fEPSPs. These results suggest that multiple mGluR subtypes (belonging to groups II and III) are expressed at mossy fiber synapses of the guinea pig hippocampus and activation of the receptors reduces the synaptic excitation, although we cannot exclude the possibility that guinea pig mossy fiber-CA3 synapses express a single class of mGluRs with unique pharmacological profiles. PMID- 8710214 TI - Influence of osmolality on seizure amplitude and propagation in the rat dentate gyrus. AB - We tested the role of changes in extracellular space on synchronization and propagation of epileptiform activity in the dentate gyrus in vitro by manipulating the osmolality of the perfusing solution. Increasing the osmolality reduced the amplitude of the population spikes and slowed the propagation of the epileptiform activity. Decreasing the osmolality had the opposite effect. Assuming there were significant changes in the size of the extracellular space, and therefore the ease with which ephaptic interactions can occur, these results support the hypothesis that ephaptic interactions play a role in the synchronization and propagation of epileptic activity in the dentate gyrus. PMID- 8710215 TI - The biosynthesis and degradation of thiamin (vitamin B1). PMID- 8710216 TI - Monoterpenoids. PMID- 8710217 TI - "Nursing power and politics". PMID- 8710218 TI - Politics and power for nurses. PMID- 8710219 TI - Nursing's agenda for health care reform: policy, politics, and power through professional leadership. AB - This article is an eye witness account of nursing's participation in the health care reform debate from 1991 to 1994. In that debate, the nursing profession achieved high visibility and recognition for the cogency of its policy positions as developed in Nursing's Agenda for Health Care Reform and for its united voice through the leadership of the American Nurses Association, the Tricouncil for Nursing, and the Nursing Organization Liaison Forum. While comprehensive health care reform failed to pass the 103rd Congress, nursing and nurses gained much in the process of their participation. PMID- 8710220 TI - Policy imperatives for nursing in an era of health care restructuring. AB - Nursing faces a markedly different health policy landscape from that which it saw during efforts for national health care reform. Sweeping changes in both the practice and political environments have required refocused policy priorities for nursing, which largely concentrate on issues of utilization of RNs and its effect on patient safety and quality of care. Nursing will need to work hard to preserve the level of unity that was achieved during health care reform but that remains critical in the face of important new challenges to the profession. PMID- 8710221 TI - "Doin' politics": linking policy and politics in nursing. AB - Policies affecting nursing are set within the larger environment of the health policy sector. That sector is characterized by increasing size, complexity, and goal displacement. All three characteristics have served to expand the resource base for this sector despite fiscal constraints on such growth. The environment of policy making shapes both the substance of policy change and the strategies for political adoption. Without understanding the dynamics of the policy environment, efforts by nursing leaders to enhance their growing political sophistication and to articulate nursing interests with maximum effectiveness will be compromised. PMID- 8710223 TI - New alliances: nursing's bright future. AB - The health care system is undergoing tremendous change. Managed care and its several incentives probably represent only an intermediate step in an unstable evolution. Equilibrium will no doubt appear with new alliances, particularly between medicine and nursing. Nurses and physicians charting a path of collaboration will ensure patients a broader set of services and high-quality care. PMID- 8710222 TI - Power in practice: a study of nursing authority and autonomy. AB - Power and politics need to be translated into practice through development of authority and autonomy within the staff nurse role. The purpose of the research described was to identify and compare aspects of agreement or disagreement between nurse leaders and staff about staff nurses' authority and autonomy to deliver patient care. Findings indicated that significant differences exist between staff nurses' and nurse leaders' sense of staff nurse autonomy, and importance of authority and autonomy, differences that can lead to serious misunderstandings and power struggles. These differences, especially concerning management support, can seriously hinder both nurse leaders and staff nurses' success in restructuring cost-effective quality care. PMID- 8710224 TI - Advanced practice politics and the Oregon nurses' trail. AB - As health care reform continues to evolve, it is important to consider the context of politics, practice, and power through an examination of nursing's recent history and participation in legislative events. Through a retrospective chronicle that includes interviews, recorded events, and article reviews, the political and legislative history of Oregon nurses in establishing advanced practice for nurse practitioners within the Oregon Nurse Practice Act is described. Prescriptive authority, hospital admitting privileges, and important elements contained in the Oregon Health Plan and Medicare reform are discussed. Questions are posed for the future of health care reform, such as the role of government in determining the quality of care in managed health care. PMID- 8710225 TI - The politics of advanced practice. AB - Nurses have a unique opportunity to influence the future. Practice opportunities abound for advanced practice nurses in emerging health delivery models. Practice barriers, external as well as those created by nursing, must be addressed. Politically savvy nurses can use their understanding of power and politics to effect changes that can maximize advanced practice nurses' contributions to the health of our society. PMID- 8710226 TI - The Connection Delivery Model: reengineering nursing to provide care across the continuum. AB - The Connection Delivery Model, a positive approach to reengineering nursing care, provides continuity across the care continuum. This model is consumer oriented and emphasizes education and prevention in providing care to high-risk, high volume patient populations. Acute care nursing staff, cross-trained to provide home care, have found this experience positive and professionally rewarding. The challenges to provide cost-effective patient care are easily met utilizing this model. PMID- 8710227 TI - Profession building in the new health care system. AB - To further the goals of greater legitimacy and power, it is necessary for nursing to abandon many of the profession building strategies it has pursued in the past. We describe the traditional framework for profession building in nursing and some reasons it is no longer effective. In contrast, a contemporary framework for profession building in nursing is presented. Because nursing, like other complex adaptive systems, is strongly anchored by its past trajectory, it will require considerable energy and resources to make the transition. PMID- 8710228 TI - Information is power. PMID- 8710229 TI - National health policy: lessons from a hot seat. AB - The author served as the nation's acquired immune deficiency syndrome (AIDS) policy coordinator from August 1993 to July 1994. Given the background of conflict surrounding human immunodeficiency virus (HIV) and AIDS policy in the United States, conflict about the position and function of a national policy coordinator is not surprising. This article discusses some of the expectations about the position, both positive and negative experiences, and lessons learned. The need for structural clarity, key staff backup, and clear agreement on timeliness is critical for anyone assuming a new, highly visible, politicized, ambiguous position. PMID- 8710230 TI - Growth in utero and coronary heart disease. PMID- 8710231 TI - 16th Marabou Symposium. Early Nutrition and Lifelong Health. PMID- 8710232 TI - Nutritional determinants of puberty. PMID- 8710233 TI - Early dietary influence on later immunocompetence. PMID- 8710234 TI - Early determinants of adult metabolic regulation: effects of infant nutrition on adult lipid and lipoprotein metabolism. AB - In a series of experiments over the past 20 years, we have demonstrated long-term deferred effects of infant nutrition, particularly breast- as compared with formula-feeding and overfeeding as compared with normal or underfeeding, on serum HDL-cholesterol concentrations, adiposity, and atherosclerosis in the baboon, a large nonhuman primate. Low HDL-cholesterol levels and obesity are associated with accelerated progression of atherosclerosis and with increased risk of coronary heart disease in humans. We have observed other deferred effects of infant nutrition on bile acid metabolism, enzyme activities, and water and electrolyte balance, some of which may be physiologically related to HDL cholesterol levels or to adiposity. The occurrence of these deferred effects suggests that infant nutrition may program other metabolic systems for life, and that these effects may contribute to other chronic diseases of adults. Although our understanding of the mechanisms by which infant diet regimens affect adult metabolism is meager, it is important to identify these mechanisms because they are likely to provide valuable clues to the causes and ultimately may contribute to the long-range prevention of those diseases. PMID- 8710236 TI - Timing and vulnerability in research on malnutrition and cognition. PMID- 8710235 TI - Early nutrition and later physical work capacity. PMID- 8710237 TI - Long-term programming of body size. PMID- 8710238 TI - Biochemical features of grain legume allergens in humans and animals. AB - Peanuts and soybeans are the major legumes involved in human food allergy, although some data exist on adverse reactions to temperate legumes including pea, green bean, sweet lupin, and lentil. An increasing number of legume proteins or glycoproteins have been characterized as food allergens. Limited data tend to indicate that they are usually different from legume inhalent allergens. Cross recognition among legume allergens is immunochemically frequent but clinically less common. A common feature to most legume allergens is their natural resistance to thermal, chemical, and in some way, proteolytic denaturation. Finally, other mammals including preruminant calves, and piglets at the time of weaning, are prone to gut immune-mediated reactions to soybean and pea proteins. PMID- 8710239 TI - The role of stereospecific saturated fatty acid positions on lipid nutrition. AB - The saturated fatty acids in the sn-1 and -3 position of triacylglycerols can exhibit different metabolic patterns due to their low absorptivity. This means that dietary fats containing saturated fats primarily in sn-1 and -3 positions (e.g., cocoa butter, coconut oil, and palm oil) can have very different biological consequences than those fats in which the saturated fats are primarily in the sn-2 position (e.g., milk fat and lard). Differences in stereospecific fatty acid location should therefore be an important consideration in the design and interpretation of lipid nutrition studies and in the production of specialty food products. PMID- 8710240 TI - Cost containment through nutrition intervention. AB - As health care cost analysts and administrators look for ways to maximize reimbursement revenues and lower operating costs without compromising the quality of patient care, reductions in nutrition services are often targeted. However, improved or expanded nutrition services can actually help cut hospital costs or increase revenues if they are targeted to patients at risk of malnutrition, applied early in the hospital course, and their benefits are appropriately documented. PMID- 8710241 TI - Nutritional and hormonal regulation of fatty acid synthase. AB - Fasting causes a decrease in the rate of synthesis of fatty acid synthase, the central enzyme in fatty acid synthesis, while refeeding carbohydrate increases synthesis. Insulin also increases the synthesis of fatty acid synthase, while glucagon causes a decline. The mechanism was shown to be transcriptional activation-mediated through a 2.1-kb stretch of the 5'-flanking sequence of the fatty acid synthase gene promoter that contains an insulin response element. These effects were confirmed by in vivo experiments with transgenic mice. PMID- 8710242 TI - Anticholesterol antibodies and plaque formation. AB - Immunization of rabbits with a protein-free formulation consisting of liposomes containing 71% cholesterol and lipid A induced cholesterol antibodies in rabbits fed a diet containing 0.5% and 1.0% cholesterol. Elevation in plasma cholesterol levels was significantly less in immunized than in nonimmunized rabbits. Immunization also resulted in a marked decrease in the risk of developing atherosclerosis as determined by analysis of aortic atherosclerosis by quantitative histological examination and fatty streaks by automated morphometric probability-of-occurrence mapping. PMID- 8710243 TI - Nurse practitioner reimbursement: history and politics. PMID- 8710244 TI - A new antihypertensive class: the angiotensin receptor antagonist. PMID- 8710245 TI - The HIV antibody test. PMID- 8710246 TI - Making oral contraceptives available over-the-counter. PMID- 8710248 TI - The relationship between parental socioeconomic levels and potential for child abuse. AB - Research studies on child abuse report on such aspects as socioeconomic levels, characteristics of abuse, demographic factors, and assessment for detection of potential abusers. Characteristics of potential or actual abusers are frequently cited in relation to low socioeconomic status. Yet at the same time, data indicates that child abuse has no socioeconomic boundaries. There is minimal data reported that would help nurses to identify potential child abusers from higher socioeconomic levels. Study is needed to identify if there are any differences in the potential for child abuse as measured between parents from low socioeconomic status and parents from high socioeconomic status. PMID- 8710247 TI - Infectious mononucleosis. AB - Infectious mononucleosis is an acute, self-limiting, nonneoplastic lymphoreticular proliferative disorder characterized by peripheral lymphocytosis and circulating atypical lymphocytes. Epstein-Barr virus is the causative agent in 90% of cases. Highest incidence is in the 15- to 25-year-old age-group, with 1% to 3% of all college students in the United States affected each year. Clinical manifestations vary according to age at presentation. Incubation period is 4 to 7 weeks. Diagnosis is primarily made with the monospot test but may include throat culture and complete blood count with differential. Cytomegalovirus and human immunodeficiency virus are among the many other conditions that may present initially as infectious mononucleosis. Treatment is supportive with prevention of complications as the goal; good personal hygiene and avoidance of contact sports should be stressed. PMID- 8710249 TI - The prevalence of smoking among women who report a history of physical and sexual abuse. PMID- 8710250 TI - Follow-up appointment compliance of discharged patients from a pediatric emergency department. PMID- 8710251 TI - Myth, mystique, and monopoly in the prescription of medicine. PMID- 8710252 TI - An investigation of the preceptor as potential mentor. PMID- 8710253 TI - The psychiatric NP in an inpatient setting. PMID- 8710254 TI - Merging advanced practice roles. PMID- 8710255 TI - Promoting the nurse practitioner by using a marketing approach. AB - Being good at what you do is not enough. Despite strong evidence that the nurse practitioner delivers cost-efficient and outcome-based care, few consumers really understand the nurse practitioner's scope of practice. With the current rapidly changing status of health care, the nurse practitioner is presented with many opportunities. Strategies that ensure survival and growth are critical to longevity. Marketing strategies can offer solutions to these challenges. The 4 P's of marketing are discussed as an approach to promote the role of the nurse practitioner. PMID- 8710256 TI - Planning for pediatric laceration repairs. AB - Primary care providers can manage the majority of pediatric lacerations presenting to a primary care setting. To facilitate planning for the care of children with lacerations, this article describes the children and lacerations who presented to an HMO pediatric urgent care setting. Most children were young, with a mean age of 6.4 years for girls and 7.4 years for boys. Most lacerations were small wounds requiring zero to five sutures. Forty-nine percent were on the face, and 97% of the wounds were repaired by primary care providers. The following issues are discussed: techniques for assessing and repairing simple facial lacerations, suggestions for helping parents and for facilitating cooperative behavior in children, how to care for an uncooperative child, criteria for referring a child to a specialist, and institutional policies that facilitate quality care for children with lacerations. PMID- 8710257 TI - Clinical diagnosis and management of the patient with deep venous thromboembolism and acute pulmonary embolism. AB - Pulmonary embolism is the third most common acute cause of death in the United States. There are approximately 500,000 cases annually in this country, leading to death in 50,000. Subjective symptoms and objective findings can oftentimes be confusing and nonspecific. A pulmonary embolism is defined as an occlusion of one or more pulmonary vessels by material that has traveled there from outside of the lung and is usually caused by a dislodged thrombus that originated in the deep veins of the legs or pelvis. Risk factors include older age, prior thromboembolism, immobility, cancer, chronic disease, congestive heart failure, pelvic and lower extremity surgery, varicosities, obesity, and oral contraception. This article will discuss current modalities that are used in the evaluation of deep venous thromboembolism and pulmonary embolism and include ventilation/perfusion scan, ultrasonography, impedance plethysmography, pulmonary angiography, and newer tests including D-dimer assays and spiral computed tomography. Medical management including simple and complex decision making, anticoagulation, and thrombolytic therapy will also be discussed. An ounce of prevention is worth a pound of gold--identification of risk factors and the use of appropriate therapeutic measures can reduce an individual's risk for deep venous thromboembolism and pulmonary embolism. PMID- 8710258 TI - Traveling healthy. PMID- 8710259 TI - A national profile of nursing centers. Arenas for advanced practice. AB - Escalating costs of health care and decreasing accessibility to care are growing dilemmas in America. Primary care is being delivered in nurse-managed clinics, or nursing centers, by nurses qualified for advanced nursing practice. The purpose of this study was to build a profile of the structure and function of nursing centers, noting major differences between those that are academic-based and those that are not. Questionnaires were developed and mailed to 234 subjects/agencies in every major region of the United States. Responses were tabulated and percentages were calculated to make comparisons and build a profile of nursing centers, based on the following factors identified in the literature and by nurse experts: (a) funding and fees, (b) organizational structure, (c) time elements, (d) demographics, (e) services, (f) qualifications and categories of staff, (g) functions of staff (h) research endeavors, (i) major barriers, and (j) major sources of support. PMID- 8710260 TI - Adult cancer detection screening by physical examination. PMID- 8710261 TI - American attitudes toward body fatness. PMID- 8710262 TI - Epinephrine. Tragic overdose. PMID- 8710263 TI - Myths & facts ... about thyroid disease. PMID- 8710264 TI - Actionstat. Bradycardia. PMID- 8710265 TI - Embolism: targeting an elusive enemy. PMID- 8710266 TI - Pancreatitis: managing a flare-up. PMID- 8710267 TI - Offering Aunt Margie comforting words. PMID- 8710268 TI - How to respond rapidly when chest pain strikes your patient has severe, terrifying chest pain. PMID- 8710269 TI - Too close for comfort: drawing the boundaries of the nurse/patient relationship. PMID- 8710270 TI - Central intravenous lines: your role. PMID- 8710271 TI - Blood a blood transfusion can be a patient's lifeline. use this guide to make sure the procedure runs safely and smoothly. transfusions: playing it safe. PMID- 8710272 TI - Assessing for ascites. PMID- 8710273 TI - Another birthday for Oscar. PMID- 8710274 TI - Making the most of every day. PMID- 8710275 TI - A letter to Mom. PMID- 8710276 TI - Positioning your patient for better breathing. PMID- 8710277 TI - Myth & facts ... about Reye's syndrome. PMID- 8710278 TI - After the fall. PMID- 8710279 TI - Performing safe finger sticks. PMID- 8710280 TI - Caring for a patient who has a vision impairment. PMID- 8710282 TI - Mother's day miracle. PMID- 8710281 TI - Warfarin. PMID- 8710283 TI - Actionstat. Ruptured ectopic pregnancy. PMID- 8710284 TI - Nosocomial pneumonia: repelling a deadly invader. PMID- 8710285 TI - Hazards of hypoxemia: how to protect your patient from low oxygen levels. PMID- 8710286 TI - Nurses week honoring those who care. PMID- 8710287 TI - Normal sinus rhythm. PMID- 8710289 TI - Peritoneal dialysis: making a clean sweep. PMID- 8710288 TI - New drugs. PMID- 8710290 TI - Judging by reactions. PMID- 8710291 TI - Caring for patients with diabetes insipidus. PMID- 8710293 TI - Self test. Reviewing the renal system. PMID- 8710292 TI - A call from the heart. PMID- 8710294 TI - Holding out hope. PMID- 8710295 TI - Hear ye, hear ye, novice and seasoned authors! PMID- 8710296 TI - Building knowledge through qualitative research: the road less traveled. AB - In this article, the author specifies the ontological, epistemological, and methodological differences among selected borrowed qualitative research methods and the three extant nursing research methods. A further distinction is made between nursing knowledge per se and general knowledge that can be used by nurses. Building nursing knowledge requires a connection with extant nursing theories and frameworks. PMID- 8710298 TI - Transcendence: formulating nursing perspectives. PMID- 8710297 TI - Workroles of staff nurses in psychiatric settings. AB - The purpose of this research was to operationalize Peplau's workroles of the psychiatric staff nurse. Thirty registered nurses audiotaped one-to-one interactions with 62 adult, child, and adolescent psychiatric patients. Content analysis was used to identify role behaviors and to identify roles that were different from those outlined by Peplau. The counselor role was the most frequently occurring primary workrole. Overlap was found between behaviors indicative of autocratic leader versus surrogate and those of resource person versus teacher. The findings supported Peplau's contention that the counselor role is central to the practice of psychiatric nursing. PMID- 8710299 TI - The articulation of nursing systems with dependent care systems of intergenerational caregivers. AB - Family members, especially adult children, provide the majority of care required by aging relatives. While there is a plethora of literature on family caregiving, generally, the intergenerational caregiving experience is not well documented. The purpose of this article is to explain the articulation of nursing systems with dependent care systems of intergenerational caregivers from three perspectives: (a) Orem's dependent care unit of service, (b) Taylor's interpretation of family as dependent care systems, and (c) Bowers' theory of intergenerational caregiving. PMID- 8710300 TI - Parse's research methodology and the nurse researcher-child process. AB - The Parse research method is a human science based nursing research methodology which views young children as partners in research about universal lived experiences. To view children in this way requires a considerable revision of the view of children assumed in many traditional theories of childhood and in research with children. In the human science paradigm, persons of all ages are viewed as human beings who make choices and assign meaning. In accord with the assumptions of the Parse research method, children are considered the experts about their health and capable of contributing to nurses' understanding of the human-universe-health process. The remarkable variation of children's thoughts, feelings and actions testifies to the complex and open nature of human becoming at all ages. Children's thoughts, feelings, and imaginings are made more accessible with the aid of art, stories, and play when offered in true presence. PMID- 8710301 TI - Challenging the myths surrounding qualitative research in nursing. AB - The challenge of nursing in the 21st century is to ground nursing practice in nursing science. This article focuses on the role qualitative research has in advancing nursing science and the myths surrounding qualitative research in nursing. The authors challenge these myths and suggest precepts to guide future research and theory development in nursing. These precepts focus on philosophical origins concerning the nature reality and the nature of knowing. Such understanding is vital to conducting and evaluating qualitative research. PMID- 8710302 TI - The conservation principles: a retrospective. PMID- 8710304 TI - The use of computers in qualitative research. PMID- 8710303 TI - Storyteller and the tall man. PMID- 8710305 TI - Ethical Narratives in practice. PMID- 8710306 TI - The importance of theory-guided research and practice in the changing health care scene. PMID- 8710307 TI - What constitutes nursing science? PMID- 8710308 TI - Canonical correlation analysis and its use in Rogerian research. PMID- 8710309 TI - A reflective moment with false cheerfulness. PMID- 8710310 TI - The human becoming theory: challenges in practice and research. AB - With rapid changes in science, technology, management, and economic planning, new challenges confront nursing. The author specifies some of the ramifications related to changes in the health care delivery system in general and in nursing in particular. The human becoming theory is discussed as a guide to nursing practice and research and is specified as a way to fortify nursing's identity. Some challenges for those living and learning the theory in practice and research are also discussed. PMID- 8710311 TI - The theory of goal attainment in research and practice. AB - King's conceptual system may be used to build a world community of nurses who respect cultural differences and share the mutual goal of health in each nation. The concepts in this systems framework have been the basis for research and practice and for developing theories such as the theory of goal attainment. This theory includes a human process of interactions that lead to transactions and to goal attainment (outcomes). This process is useful in any nursing situation and in any culture. Knowledge of the concepts of the theory have been applied in practice in the United States, Canada, Sweden, and Japan. An example of implementing theory-based practice in a hospital is discussed. PMID- 8710312 TI - The Neuman systems model in research and practice. AB - The major focus of this article is on utilization of the Neuman systems model for nursing practice and research. The directives given will accommodate cultural variations and also serve the needs of interdisciplinary groups. The author shares views of global values for using the Neuman systems model for relevant nursing education, practice, administration, research, and international and interdisciplinary functions. PMID- 8710313 TI - Culture care theory, research, and practice. AB - Today nurses are facing a world in which they are almost forced to use transculturally-based nursing theories and practices in order to care for people of diverse cultures. The author, who in the mid-50s pioneered the development of the first transcultural nursing theory with a care focus, discusses the relevance, assumptions, and predictions of the culture care theory along with the ethnonursing research method. The author contends that transcultural nursing findings are gradually transforming nursing practice and are providing a new paradigm shift from traditional medical and unicultural practice to multiculturally congruent and specific care modalities. A few research findings are presented to show the importance of being attentive to cultural care diversities and universalities as the major tenets of the theory. In addition, some major contributions of the theory are cited along with major challenges for the immediate future. PMID- 8710314 TI - Thank you, Dorothy: Now I know. PMID- 8710315 TI - Graduate professional development programme dentistry. AB - This paper describes the development and structure of a pilot programme of professional development for new graduates initiated by the New Zealand Dental Association, the Dental Council of New Zealand, and the Faculty of Dentistry of the University of Otago. The programme consisted of a series of monthly evening seminars, co-ordinated by a facilitator, and led by experienced practitioners. Initially provided in Auckland, the programme subsequently extended to Wellington, Christchurch, and Waikato-Bay of Plenty. The award of a New Zealand Dental Association Travelling Fellowship enabled the author to observe the United Kingdom Vocational Training Scheme, and attendance at the 1994 meeting of the Federation Dentaire Internationale provided opportunity to study systems used in other countries. The pilot programme has now been replaced by the ongoing Graduate Professional Development Programme Dentistry, a programme which extends beyond the recent graduate, but remaining based on the concepts developed in the pilot programme. PMID- 8710316 TI - A combined endodontic-periodontal lesion of development origin: a case report. AB - This report describes an instance in which treatment of a maxillary lateral incisor tooth failed to resolve the presenting endodontic-periodontal lesion. The predisposing aetiological factor was a longitudinal developmental groove extending from the cingulum to the apex of the tooth. PMID- 8710317 TI - Establishing a competitive advantage through quality management. AB - The successful dentist of the future will establish a sustainable competitive advantage in the marketplace by recognising that patients undergoing dental treatment cannot see the result before purchase, and that they therefore look for signs of service quality to reduce uncertainty. Thus the successful dentist will implement a quality programme that recognises not only that quality is defined by meeting patients' needs and expectations, but also that quality service is fundamental to successful business strategy. Finally, the successful dentist of the future will realise that the pursuit of quality is a never-ending process which requires leadership by example. PMID- 8710318 TI - Corporate liability: security and violence--Part II. AB - A hospital can be held liable for injuries resulting from failure to provide adequate, reasonable security Part II of "corporate Liability: Security and Violence" addresses negligent hiring and supervision practices, injury and domestic violence in the workplace and communication procedures. PMID- 8710319 TI - The pulse of managed care in 1996 and beyond. AB - The key to success in any managed care arrangement is building relationships between the players. Thus, it is imperative that the subacute care provider develop and nurture positive working relationships with MCOs and other sources of referrals. It is no longer enough to have an MCO contract. It is what happens after the contract has been signed that makes a difference on whether or not a subacute care provider can thrive in the managed care environment. PMID- 8710320 TI - Creating a true learning organization. AB - Where possible, an environment should be composed of opportunity and excitement, rather than fear of failure. New models of behavior transform fear of failure without losing the overarching commitment to the patient. PMID- 8710321 TI - The integral role of nursing in managed care. AB - Implementing a managed care model into the ambulatory care setting results in a user-friendly climate. A primary care treatment team coordinates all the patients' health care needs. Goals are to assign every patient to a specific attending physician and to integrate both inpatient and outpatient services into a managed care environment. PMID- 8710322 TI - Family Medical Leave Act: ready or not! AB - Many employers already have policies in place that grant job leaves for a variety of reasons. It is no longer optional for employers to grant job leaves; appropriate leave is now the law. PMID- 8710323 TI - Improving documentation of initial nursing assessment. AB - Greater efficiency and ease of use results when nurses use an admission assessment form that is integrated with patient care outcome standards. Steps in developing such a focus are described. PMID- 8710324 TI - MERG: medication event rating grid. AB - Through a four-step process, a medication event rating grid (MERG) tracks medication errors. An educational and a remedial plan are included to help individual staff members develop strategies to prevent future errors. PMID- 8710325 TI - Long-term pediatric care in the managed care environment. AB - Pediatric subacute care is a transitional phase along the continuum of care. Consumers, providers and payors must address the system-wide changes needed to integrate this population within the subacute care environment. PMID- 8710326 TI - Improving admission and discharge: quality improvement teams. AB - A brief pilot study resulted in significant reduction in admission and discharge times at a large Florida hospital. This was achieved by changing the work hours of the day shift environmental service staff. PMID- 8710327 TI - Creation of a new order. AB - The underlying values of AONE include health, knowledge/discovery/insight, interdependence, stewardship and creation of a new order. Research, definition, reflection and action help to achieve this vision. PMID- 8710328 TI - Teaching effective communication skills. AB - Fostering adaptive communication skills through development, initiation and maintenance of an ongoing staff education program is presented. Goals of the proposed interventions include: developing functional interpersonal relationships, increasing job satisfaction and decreasing perceived levels of work-related stress among staff nurses. PMID- 8710329 TI - TQM: pressure ulcer prevention. AB - An interagency task force has designed and implemented a pressure ulcer prevention program at a county hospital. PMID- 8710330 TI - Manage authorizations to manage care. AB - Managed care organizations rely on different ways to authorize the utilization of hospital services, physician specialty care and other referral services. Critical differences revolve around the role the primary care physician plays in the authorization process. The more tightly managed the system, the more likely will unnecessary medical costs be avoided. PMID- 8710331 TI - Laparoscopic cholecystectomy: a technologic assessment. AB - Laparoscopic cholecystectomy, despite its short clinical history, has clearly established itself as the preferred surgical therapy for gallbladder removal in symptomatic patients. Significant questions remain, however, regarding the appropriate credentialing criteria for surgeons performing these procedures, the potential incidence of long-term complications, therapeutic guidelines for the assessment and intervention with patients with common duct stones and those select patient subgroups that can be optimally managed with medical, rather than surgical, therapy. The technology assessment team plays a critical role in helping to design mechanisms to answer these important outcomes inquiries. PMID- 8710332 TI - Managed care of the short-stay obstetric patient. PMID- 8710333 TI - The Malcolm Baldrige Health Care Pilot. PMID- 8710334 TI - Recruitment and retention in subacute care. PMID- 8710335 TI - Beastly behavior. PMID- 8710336 TI - New national sample survey of registered nurses. PMID- 8710337 TI - Is a medical savings account in your future? AB - Proposed federal legislation would expand the coverage options Medicare beneficiaries have. One option--Medisave--would enable beneficiaries to replace traditional Medicare coverage with a catastrophic insurance policy coupled with a medical savings account. At least initially, this risky insurance product is likely to appeal to only a small segment of the Medicare population. PMID- 8710338 TI - The challenges and pitfalls of a managed care contract. AB - Recognizing that managed care contracting is both an art and a science no longer suffices in this era of merger mania and integrated delivery networks. Subacute care providers must now strategize to meet the challenges posed after a managed care contract has been fully executed. Equally important, the pitfalls of a managed care contract must be identified to avoid serious negative consequences. PMID- 8710339 TI - Law for the nurse manager. Legal update, 1995: Part 1. AB - As the health care system undergoes change, the traditional areas of liability continue to exist while new areas of risk exposure evolve. Trends are identified. PMID- 8710340 TI - A guide on integrating clinical information. AB - Clear action plans can guide nurse executives toward creating an integrated clinical environment. A guide identifies how each environment is affected by changing delivery models. PMID- 8710341 TI - The vision of LTC in state veterans facilities. AB - Four state-operated veteran facilities adopted a vision to ensure consistency. The core of the vision established a complimentary, state-wide system of care to meet all the members' diverse needs. PMID- 8710343 TI - A high-involvement health care model. AB - Integrating a traditional approach into a high-involvement culture can result in successful redesign. Examples of successful redesign efforts as well as those that failed are discussed. PMID- 8710342 TI - The connection delivery model: care across the continuum. AB - Cross-training nursing personnel to provide follow-up home care to patients with congestive heart failure has resulted in a dramatic drop in readmissions. Significant reduction in length of stay and cost for readmissions has prompted this hospital to expand this type of service to other patient populations. An added benefit is high job satisfaction among nurses. PMID- 8710344 TI - Work empowerment and organizational commitment. AB - As organizations struggle to deliver the same level and quality of services with fewer resources, administrators are challenged with redesigning workplaces to maximize nurses' commitment. This study used Kanter's Structural Theory of Organizational Behavior to examine the relationship between job-related empowerment perceptions of staff nurses and their commitment to the organization. Strategies for creating more empowered work environments are discussed. PMID- 8710345 TI - A decision-making process in shared governance. AB - Shared governance teams require a good decision making process. After identifying the root cause of a problem, finding a solution that addresses the right issues begins. An issues flow chart and decision-making process were developed to help teams better identify and solve problems. PMID- 8710346 TI - Stat nursing--the final analysis. AB - Stat nursing was designed to alleviate short-term workload fluctuations occurring in intensive care units (ICUs). The stat nurses (SNs) "rove" throughout the hospital, using a priority schedule to intervene in short-term staffing crises that include, but are not limited to, code blues, ICU postoperative recoveries, ICU patient transports for radiological procedures, ICU patient admissions, ICU bedside procedures, technical consultation, trouble-shooting equipment and i.v. therapy. PMID- 8710347 TI - Changing cost-accounting procedures in the ED. AB - A study addressing cost containment in the emergency department examines nurses' attitudes toward cost-effectiveness and their reasons for noncompliance. Strategies to improve cost-accounting procedures are given. PMID- 8710348 TI - Perioperative introductory column. AB - Our future in the operating room (OR) depends on our ability to remain productive and valuable to surgeons, patients and administration. Together, we can ensure that quality and efficiency in the OR are not casualties of the health care revolution. Let us all share our knowledge and experience to help ourselves through to the next century. PMID- 8710349 TI - Chest pain observation units. AB - The key to technologic assessment of the chest pain unit lies in the recognition of the unit's ability to optimize outcomes of an illness with high incidence, high acute mortality and significant chronic morbidity. The chest pain observation unit, as an adjunct to the emergency department (ED), offers health managers and administrators the clinical tool, first envisioned by Dr. Lown in the 1960s, to achieve improved quality and cost-effectiveness of acute chest pain management as health care moves into a new century. PMID- 8710350 TI - First you suffer, then you die: findings of a major study on dying in U.S. hospitals. AB - A recent study of 9,000 dying patients found that improving communications between physicians, patients and families did not improve the quality of the care the patients received. Though provided with up-to-date information about patient prognoses and levels of pain, physicians made no change in their treatment patterns. This article reviews commentators from medicine, philosophy and the media and suggests actions that might improve care of the dying. PMID- 8710351 TI - Making a change: the importance of administrative support. PMID- 8710353 TI - Successful Restructuring. PMID- 8710352 TI - "Manager as developer": facilitating collaboration. PMID- 8710354 TI - Other people's lives ... PMID- 8710355 TI - OH: a jack of all trades? PMID- 8710356 TI - Breast cancer awareness. PMID- 8710357 TI - Fence building: providing emotional support. PMID- 8710358 TI - Building a dream team. PMID- 8710360 TI - AIDS projections. PMID- 8710359 TI - Disability, discrimination and the law. PMID- 8710361 TI - One billion years of Myb. AB - The v-myb oncogene of the avian myeloblastosis virus has led to the discovery of a large and growing family of myb-related genes in a wide variety of eukaryotes including animals, plants, fungi and slime molds. The Myb-related proteins contain a highly conserved sequence, often present in multiple tandem repeats which constitute a DNA-binding domain. These proteins generally function in the regulation of cell growth and differentiation, often by coregulating gene expression along with DNA-binding proteins of other classes. This review focuses on the evolution of the myb gene family and the role of these genes in development. PMID- 8710362 TI - G1 cyclin/CDK-independent phosphorylation and accumulation of p130 during the transition from G1 to G0 lead to its association with E2F-4. AB - During the transition from G1 to G0, p130 undergoes a specific phosphorylation event-leading to p130-form 2- that is mediated by a kinase/s other than the known G1, S and G2/M cyclin/CDKs. Changes in the phosphorylation status of p130 during this transition are responsible, at least in part, for the concomitant formation of p130/E2F-4 complexes, which are characteristic of G0. These complexes remain abundant during early G1 upon restimulation, but not after mitosis, and are dissociated in mid G1 when p130 is abruptly hyperphosphorylated to form 3. Subsequently, p130 forms 1 and 2 are no longer detected during the remainder of the cell cycle. Consistently, phosphorylation to form 3 and dissociation from E2F 4 complexes is reproduced by a cyclin/CDK holoenzyme in vitro. TGF-beta-induced G1 arrest abrogates cyclin/CDK phosphorylation of p130 but not phosphorylation to form 2. The cell cycle-dependent phosphorylation pattern of p130 is thus shown to comprise two distinct steps that are catalyzed by different kinases. The differential regulation of p130 and pRB phosphorylation during the transition from G1 to G0 may explain the fact that p130 and E2F-4 are the major components of E2F complexes in quiescent cells. Moreover, the newly described phosphorylation of p130 at the transition from G1 to G0 defines a novel mechanism of cell cycle exit regulation. PMID- 8710363 TI - Constitutive activation of STAT5 by the BCR-ABL oncogene in chronic myelogenous leukemia. AB - Using chronic myelogenous leukemia (CML) as a model, we tested the hypothesis that cytokine-independent growth of leukemia cells results from aberrant activation of cytokine signaling pathways. The STAT5 (signal transducer and activator of transcription) protein, which is activated transiently in normal myeloid cells by cytokines such as GM-CSF (granulocyte-macrophage colony stimulating factor), was constitutively activated in cell lines derived from CML patients, even in the absence of GM-CSF. STAT5 was also activated in primary mouse bone marrow cells acutely transformed by the CML-specific BCR-ABL oncogene, but not by the serine kinase oncogene v-MOS. Reconstitution experiments in non hematopoietic cells show that STAT5 activation by BCR-ABL occurs independent of cytokines. Results using BCR-ABL mutants which specifically uncouple connections to known signal transduction pathways show that STAT5 activation is kinase dependent and correlates directly with ability to confer cytokine independent growth in hematopoietic cells. BCR-ABL also activates JAK kinases, which may provide a mechanism for STAT activation. These findings are consistent with a role for STAT5 in hematopoietic transformation by BCR-ABL. PMID- 8710364 TI - Characterization of ES cells deficient for the p105 precursor (NF-kappa B1): role of p50 NLS. AB - Proteolytic processing of the p105 precursor (NF-kappa B1) generates the p50 subunit of NF-kappa B. To address the function of p105 precursor, we have generated embryonic stem (ES) cells which lack the C-terminal inhibiting ankyrin containing domain of p105 (I kappa B gamma) but contain a functional p50 by homologous recombination. Two forms of p50 were found in ES cells lacking p105. While one was a p50 molecule with the expected size of 415 amino acids, the other was an isoform of 358 amino acids generated by alternative splicing. This isoform lacked the C-terminal 62 amino acids of p50 including the putative nuclear localization signal (NLS), but included five unrelated amino acids. This p50 molecule lacking the NLS behaved similar to wild type p50. Importantly, the p50 isoform-RelA complex interacted efficiently with I kappa B alpha, implying that the NLS of p50 is not essential for these functions. Also, p50 isoform dimers were present in the nucleus. Loss of p105 and the augment of total p50 produced an increase in the nuclear kappa B-binding activity of p50-RelA and p50 dimers. In wild type ES cells, a significant proportion of total p50 is associated with p105. Therefore, the mutation introduced in NF-kappa B1 eliminates both the processing to generate p50 and the trapping of p50 by p105. In contrast to p50, only a small fraction of RelA is associated with p105. These findings indicate that p105 plays a unique role in controlling p50 dimer activity. PMID- 8710365 TI - Induction of apoptosis by p53 is independent of its oligomeric state and can be abolished by HPV-18 E6 through ubiquitin mediated degradation. AB - Inhibition of p53 function is a common feature of many DNA tumour viruses. Human papillomavirus (HPV) E6 proteins from the oncogenic HPVs inhibit p53 function either by blocking its ability to bind DNA or by labelling newly synthesised p53 as a target for ubiquitin mediated degradation. In this study we have investigated the role of the degradation function of E6 with respect to p53 function. Using a panel of previously characterised p53 mutant proteins we have been able to establish a series of assays which separates p53 growth suppression from transformation suppression and from induction of apoptosis. Only wild type p53 inhibits the growth of p53 null 10(1) cells, whereas wild type, dimeric and monomeric mutants of p53 suppress transformed cell growth of both Saos-2 cells and baby rat kidney cells. Cells expressing the different oligomeric forms of p53 all retain the ability to induce apoptosis upon u.v. treatment. Using HPV E6 and E7 we have been able to show that E7 will overcome p53 growth suppressor activity with an efficiency similar to that observed with E6. However, in contrast to E6, E7 has no effect on the ability of p53 to suppress transformed cell growth. Finally, we show that the ability of E6 to label p53 for ubiquitin mediated degradation is prerequisite for its ability to overcome p53 inhibition of transformed cell growth and induction of apoptosis. These observations argue that E6 inhibits p53 mediated apoptosis and suppression of transformation while E7 inhibits p53 suppression of cell proliferation. PMID- 8710366 TI - Intracellular expression of a single-chain antibody directed to the EGFR leads to growth inhibition of tumor cells. AB - A gene encoding a single-chain antibody (scFv) which specifically binds the epidermal growth factor receptor (EGFR) has been constructed from hybridoma cells producing the R1 monoclonal antibody. The gene, designated scFv-R1R, was introduced into EGFR transformed NIH3T3 cells via retroviral infection. scFv-R1R was directed to the lumen of the endoplasmic reticulum (ER) where it bound the extracellular domain of the receptor inhibiting its appearance on the plasma membrane. In these cells, EGF induced tyrosine phosphorylation of the EGFR and several substrates was greatly reduced. Furthermore, intracellular retention of EGFR caused a partial inhibition in the transformed growth of the cells. Intracellular expression of receptor tyrosine kinase directed scFvs is a novel approach for affecting tumor cell growth. We have recently shown that scFv-5R directed to ErbB2, another member of the ErbB family, blocks the anchorage independent growth of ErbB2 transformed cells. In order to examine the effects of scFv-R1R and scFv-5R on the long-term growth of tumor cells overexpressing either EGFR or ErbB2, retroviruses encoding the two scFvs were used to infect various human tumor cell lines. Intracellular expression of the scFvs resulted in a marked inhibition of stable colony formation in some of the cell lines. In general, inhibition was observed when overexpressed receptor was targeted. However, in some cases expression of both scFvs was incompatible with long term cell growth suggesting that heterodimers of ErbB2 and EGFR are essential for the growth of some human tumor cell lines. PMID- 8710367 TI - Isolation of novel, transcriptionally active AP-1 binding sites: implications for cellular transformation. AB - Increased AP-1 DNA-binding activity, in the context of TRE-binding, is not a consequence of Fos transformation. In this report we investigate the possibility of a change in binding site preference by vFosAP-1 compared with AP-1 from an untransformed cell. Fos binding sites were immunoselected from random sequence oligonucleotides using a pan Fos anti-serum with nuclear protein from quiescent FBRp75v-fos-transformed (FBR) and normal (208F) rat fibroblasts. The selected oligonucleotides were aligned by computer and a consensus described for the sequences bound by AP-1 from the two cell lines. The vFos binding site is shown to be a consensus TRE, whereas the sequence ACCACATC is described as the cellular Fos protein family consensus. We demonstrate that sequences differing from the TRE consensus can bind AP-1 and direct transcription. AP-1 DNA-binding activity differs between normal and transformed cells with several of the selected oligonucleotides. These sequences also demonstrate differential transcriptional activation between normal and transformed cells. In particular, the 208F consensus has no transcriptional activity in FBR cells. Further, EGF differentially influences the transcriptional activity of the oligonucleotides in 208F and FBR cells. Our results suggest that AP-1 may change its preferred binding site depending on the proteins available at any given time, the sequences flanking a non-consensus TRE or even the environment in which the cell exists. These differences in binding site preference and transcriptional activation may result in the increased transforming ability of the v-fos oncogene compared with the c-fos proto-oncogene and may extend the potential target genes beyond those with an AP-1 consensus binding site. PMID- 8710368 TI - p53 binds to a novel recognition sequence in the proximal promoter of the rat muscle creatine kinase gene and activates its transcription. AB - The rat muscle creatine kinase (CKM) gene promoter is unusual since it is one of the few cellular promoters containing a p53 response element which is located proximally (bp -168 to -57) to the transcription start site. We have previously shown that p53wt transactivates transcription in vivo of rat CKM, in CV-1 monkey kidney cells, through this 112 bp promoter-proximal fragment which contains at least five degenerate p53-binding elements. In this report, we employed the gel shift assay and demonstrated that recombinant, immunoaffinity-purified mouse p53wt binds to this 112 bp CKM sequence and activates the in vitro transcription of the proximal CKM promoter by nuclear extracts from CV-1 cells. Also, a competitor plasmid containing this 112 bp CKM fragment interferred with the in vivo transactivation of CKM by p53. This CKM fragment, when cloned upstream of the rat brain creatine kinase (CKB) promoter, mediated the p53 transactivation of CKB. Analyses of p53wt and a series of missense mutants (altered in conserved region II of p53) showed that binding of p53 to the CKM promoter was required but was not sufficient for transactivation. The results are discussed in relation to the possible role of p53wt in the expression of CKM in cell types which may not express the myogenic transcription factors. PMID- 8710370 TI - Mechanisms of transcriptional transactivation and restriction of human immunodeficiency virus type I replication in an astrocytic glial cell. AB - Unique transcriptional transactivation by the human immunodeficiency virus type 1 (HIV-1) Tat protein of long terminal repeat (LTR)-driven RNA expression, in the absence of the transactivator responsive element (TAR), was previously demonstrated in central nervous system (CNS)-derived astrocytic cell-lines, including U87MG. In the present study, RNase protection assays were utilized to reveal the molecular mechanism(s) underlying transactivation of the HIV-1-LTR in these cells. Short transcripts, which represent abortive HIV-1 transcription, could not be detected either in the absence or presence of Tat, and no differences in transcript levels were detected using 5' probes, as compared to 3' probes, in the experiments. Thus, the transactivational effects of Tat, in U87MG cells, were potentially based on the increase of transcriptional initiation, both in TAR-dependent and -independent states. Further, by using newly established stable cellular transformant, containing HIV-1-LTR-reporter gene constructs, TAR independent transactivation was demonstrated to efficiently function primarily in transiently-transfected U87MG cells. U87MG cells, stably-transfected with the intact HIV-1 proviral genome, produced very low levels of virus after long-term culture, as previously reported in other astrocytic cells. These cells demonstrated profoundly restricted transcription of the HIV-1 genome, with no detectable levels of HIV-1-specific RNA by Northern blotting, indicating that the restriction of viral production in these cells is principally due to the low level of overall transcription from the 5' HIV-1-LTR. Transcription of HIV-1 RNA in this cell could not be significantly up-regulated by various stimulators, such as phorbol 12-myristate 13-acetate (PMA), tumor necrosis factor-alpha (TNF-alpha) and sodium butyrate. These data suggest that the restriction of HIV-1 transcription in these cells may be controlled by different mechanism(s) from those in lymphocytic or monocytic cells. PMID- 8710369 TI - AML-2 is a potential target for transcriptional regulation by the t(8;21) and t(12;21) fusion proteins in acute leukemia. AB - AML-1B is targeted directly and indirectly in multiple chromosomal translocations in myeloid and B-cells. The AML-1/ETO and TEL/AML-1 fusion proteins, created by the t(8;21) and t(12;21) respectively, disrupt AML-1B-dependent transcription. Recently, two human members of the runt homology domain family of transcription factors have been identified, AML-2 and AML-3, which also regulate transcription through enhancer core motifs. If multiple factors regulate transcription through the same site, a dominant interfering protein may be required to promote leukemogenesis, rather than the inactivation of both AML1 alleles. To determine which AML family proteins are active in hematopoietic cells, we developed antisera specific to each family member for use in gel mobility shift assays. We have found that AML-1B is the major DNA binding activity in T-cell lines, while both AML-1B and AML-2 are expressed in myeloid and B-cell lines. AML-1B represents most of the active protein in the mouse thymus, whereas AML-1 and AML 2 are equally expressed in the mouse spleen. AML-3 is expressed at very low levels in a single myeloid cell line, 32D.3, and is the only core binding activity present in Buffalo rat liver cells. We demonstrate that AML-2-dependent transactivation mediated by enhancer core motifs is inhibited by the AML-1/ETO and TEL/AML-1 fusion proteins. This indicates that the t(8;21) and t(12;21) fusion proteins inhibit transcriptional activation by the AML-1 transcription factor family, and in so doing contributes to leukemogenesis. PMID- 8710371 TI - Interaction of human polyomavirus BK with the tumor-suppressor protein p53. AB - We have been studying the interaction of the oncogenic human polyomavirus BK (BKV) with the tumor-suppressor protein p53 to understand the biology of this virus as well as to understand the basic mechanisms of p53 transactivation. We here demonstrate that p53 binds specifically to the viral promoter at two different sites, S-I (np 361-383) and S-II (np 314-336) in the late region. Site S-I is a 23 bp domain comprising an unique combination of a 10 bp consensus monomer binding site (Pu Pu Pu C (A/T) (T/A) G Py Py Py) which is contiguous with a GC-rich Sp-1 motif that binds p53 in the SV40 promoter. Site S-II also spans a 23 bp sequence containing two tandem consensus binding sites with three base pair mismatches in each and a one base pair deletion. A dimer of a 100 bp region spanning both the binding sites or the site S-I alone induced p53 responsiveness to a basal promoter when cloned upstream from the TATA box, but a similar construct using S-II did not. One tumor-derived mutant protein, p53-175 H, which is defective in DNA binding, also failed to transactivate the reporter gene. We further show that p53 binding-dependent transactivation is abrogated by BKV large T antigen, thereby suggesting an interaction between these two proteins in vivo. In contrast to the isolated p53 binding site, viral early promoter is repressed by p53 in H 1299 cells and the mutants are defective in this function to varying extent. This is suggestive of an involvement of cellular factors in modulating p53's function in the context of the whole promoter. p53 binding sites in BKV are flanked by the binding sites for transcription factors Sp-1 and NF-1 and we show that these transcription factors are present in the immunocomplex with purified p53, implicating modification of p53's transactivation function by protein protein interaction. Thus, oncoprotein synthesis in this virus might be modulated by p53 in vivo by a complex mechanism other than simple DNA binding and sequestration of the TATA binding protein. Together with SV40 and polyomavirus, which also harbor p53 binding sites, this viral system will serve as a model to understand the role of p53 in viral infection. PMID- 8710372 TI - Properties of Wnt-1 protein that enable cell surface association. AB - Members of the highly conserved Wnt gene family serve key functions in cellular growth and differentiation. Regulated expression of the murine Wnt-1 proto oncogene is essential for normal development of the embryonic nervous system and when misexpressed in the mammary gland leads to hyperplasias and tumorigenic progression. Wnt-1 encodes a cysteine rich glycoprotein that is secreted and tightly associated with the cell surface and/or extracellular matrix. This restricted diffusion of Wnt-1 protein is important for specifying a spatially defined pattern of Wnt-activity necessary for cell to cell signaling in cell growth and differentiation. Since the receptors and cell surface molecules that bind Wnt proteins are as yet unknown we have sought to define the sequences within Wnt-1 protein that enable cell surface association and to define the nature of the cell surface structures with which Wnt-1 protein associates. First, site directed mutagenesis was used to identify a basic amino acid sequence motif within Wnt-1 protein that is required for this protein to accumulate on the cell surface. Second, wild type Wnt-1 protein was expressed in a series of proteoglycan-deficient CHO cell lines to show that, contrary to expectations, cellular glycosaminoglycans are not required for processing, secretion, cell surface association and accumulation of Wnt-1 protein. PMID- 8710373 TI - Growth factor-dependent survival of rodent fibroblasts requires phosphatidylinositol 3-kinase but is independent of pp70S6K activity. AB - A variety of mammalian cells undergo apoptosis when deprived of growth factors, but the intracellular signaling pathways responsible for cell survival remain to be characterized. In the present study, we have investigated the role of PI 3 kinase and pp70S6K in growth factor-dependent survival of rodent fibroblasts. As previously reported for PC12 pheochromocytoma cells, Rat-1 and REF52 cells underwent apoptosis following either serum-deprivation or treatment with the PI 3 kinase inhibitors wortmannin and LY294002. In contrast, NIH3T3 and BALB 3T3 cells were resistant to apoptosis induced by either serum-deprivation or PI 3-kinase inhibition. It thus appears that PI 3-kinase is specifically required to prevent apoptosis of fibroblast cell lines that are dependent upon growth factors for their survival. Consistent with this role of PI 3-kinase, serum and growth factors maintained steady state levels of PI 3-kinase activity that rapidly decreased following serum-deprivation. Serum and growth factors similarly maintained a steady state level of pp70S6K, which is thought to be activated downstream of PI 3-kinase. However, inhibition of pp70S6K activation by rapamycin failed to induce apoptosis in either Rat-1 or PC12 cells. Cell survival thus appears to require a PI 3-kinase signaling pathway that is independent of pp70S6K activation. PMID- 8710374 TI - RalGDS-like factor (Rlf) is a novel Ras and Rap 1A-associating protein. AB - The small GTPase Rap 1A is a close relative of Ras that, when overexpressed, is able to revert oncogenic transformation induced by active Ras. We screened a mouse embryonic cDNA library using the yeast two-hybrid system and isolated the cDNA of a novel Rap 1A-interacting protein. The open reading frame encodes for an 84 kDa protein with a Cdc25-homology domain which shares approximately 30% identity with Ral guanine nucleotide dissociation stimulator (RalGDS) and RalGDS like (Rg1). The C-terminal region reveals a striking conservation of sequences with the Ras-binding domain of RalGDS. We designated this protein Rlf, for RalGDS like factor. In the yeast system, Rlf interacts with Rap 1A, H-Ras and R-Ras, but not with Rac and Rho. In addition, we found that Rlf interacts with Rap 1Aval12 but not with Rap 1AAsn17. In vitro binding studies revealed that a C-terminally located 91 amino acid region of Rlf is sufficient for direct association with the GTP-bound form of Ras and Rap 1A. The observed dissociation constants are 0.6 microM and 0.4 microM, respectively. No significant association with Ras-GDP or Rap 1A-GDP could be detected. These binding characteristics indicate that Rlf is a putative effector for Ras and Rap 1A. PMID- 8710375 TI - Isolation and characterization of murine vav2, a member of the vav family of proto-oncogenes. AB - We describe the isolation and characterization of a cDNA encoding murine vav2. vav2 shares 63% and 55% identity at the nucleic acid and amino acid levels, respectively, with vav, a proto-oncogene that plays an essential role in embryonic development and hematopoietic signal transduction. The 100 kDa Vav2 protein contains the characteristic array of structural motifs found in Vav. However, unlike vav, vav2 transcripts are widely distributed in both hematopoietic and non-hematopoietic tissues. In the adult, vav2 mRNA is found at high levels in the spleen, liver, testes and placenta. Northern blot analysis reveals two vav2 mRNA species (designated alpha and beta). The alpha species is expressed throughout development while the alpha and beta species are expressed tissue-specifically in adults. Transfection of NIH3T3 cells with expression vectors containing vav2 deletions demonstrate that elimination of 183 amino terminal residues of Vav2 is sufficient to activate its oncogenic potential. Vav2 induced transformation is characterized by the appearance of foci composed of cells in which cytokinesis and karyokinesis are uncoupled. This phenotype is comparable, but not identical, to morphological changes induced by Vav and other members of the DbI family of oncoproteins. Our results suggest that Vav family members mediate functions important in the regulation of cell architecture and proliferation in most, if not all, tissues. PMID- 8710376 TI - Repression of apoptosis in human B-lymphoma cells by CD40-ligand and Bcl-2: relationship to the cell-cycle and role of the retinoblastoma protein. AB - Using a Burkitt lymphoma cell line to model human B-cell apoptosis in vitro, we observed that crosslinking, by antibody, of cell surface immunoglobulin induced G1 growth-arrest followed by apoptosis. By contrast, cells treated with the Ca(2+)-ionophore, ionomycin, generated apoptotic signals in G2/M as well as in G1. Both ionomycin and anti-immunoglobulin treatment induced rapid dephosphorylation of Rb prior to apoptosis. Apoptosis was repressed following exposure to CD40-ligand and was accompanied by hyperphosphorylation of Rb and cell-cycle progression but not Bcl-2 expression. Expression of Bcl-2 protein in stable bcl-2-transfectants, also resulted in repression of apoptosis and anti immunoglobulin-treated cells no longer underwent growth-arrest. In Bcl-2 expressing cells in which apoptosis was repressed, Rb remained hyperphosphorylated, even during G1-arrest induced by ionomycin. TGF beta treatment of Bcl-2-expressing cells induced G1-arrest, de-phosphorylation of Rb and apoptosis. These results suggest that the functional activity of Bcl-2 in B lymphoma cells is dependent upon, or leads to, sustained hyperphosphorylation of Rb and that Rb hyperphosphorylation can be uncoupled from cell-cycle progression. PMID- 8710377 TI - Formation of Shc/Grb2- and Crk adaptor complexes containing tyrosine phosphorylated Cbl upon stimulation of the B-cell antigen receptor. AB - B-cell antigen receptor (BCR) stimulation induces tyrosine phosphorylation of the Shc adaptor protein and its association with Grb2. The Shc/Grb2 complex may be involved in Ras activation, since Grb2 interacts with the guanine nucleotide exchange factor Sos. We reveal here an additional complexity of the BCR-induced Shc/Grb2 complex: it contains tyrosine phosphorylated proteins of 130, 110 and 75 kDa. The 130 kDa molecule inducibly associates with Shc, while the 75 kDa protein interacts with the carboxy-terminal SH3 domain of Grb2. The 110 kDa molecule is defined as Cbl, the product of the c-cbl oncogene, which is strongly phosphorylated on tyrosine upon BCR stimulation. Cbl constitutively interacts with the SH3 domains of Grb2, with a preference for the amino-terminal domain, and is in this way recruited to Shc upon BCR stimulation. Immunodepletion studies showed that Grb2-associated Cbl can be phosphorylated by BCR-induced tyrosine kinases independent of a Shc/Grb2 interaction. This indicates that the BCR can also couple to a Grb2 complex without the involvement of Shc. Cbl not only interacts with Grb2, but also with the adaptor protein Crk. In contrast to its constitutive interaction with Grb2, tyrosine-phosphorylated Cbl only associates with Crk after BCR stimulation. In summary, we observe that the BCR activates Shc/Grb2-, Grb2- and Crk adaptor complexes of distinct composition, which may allow selective coupling to different signal transduction cascades. Cbl participates in all three adaptor complexes and is tyrosine phosphorylated upon BCR stimulation, pointing to a central role for this molecule in the regulation of antigen receptor-induced B cell responses. PMID- 8710378 TI - p21WAF1/CIP1 interacts with protein kinase CK2. AB - p21WAF1/CIP1 which belongs to a class of regulatory proteins that interact with cyclin dependent kinases is a potent inhibitor of these kinases. The inhibition of the cyclin dependent kinases induces an arrest of cells in the G phase of the cell cycle. In addition p21WAF1/CIP1 associates with PCNA and inhibits DNA replication. Here, we show that p21WAF1/CIP1 binds to the regulatory beta-subunit of protein kinase CK2 but not to the catalytic alpha-subunit. Binding of p21WAF1/CIP1 down regulates the kinase activity of CK2 with respect to the phosphorylation of the beta-subunit of CK2, casein and the C-terminus of p53. This study demonstrates a new binding partner for the regulatory beta-subunit of protein kinase CK2 which regulates the activity of the holoenzyme. PMID- 8710380 TI - A novel p53 germline alteration identified in a late onset breast cancer kindred. AB - Germline mutations in the p53 tumor suppressor gene are associated with the Li Fraumeni syndrome, characterized by childhood sarcoma, leukemia and early onset breast cancer and has occasionally been found also in familial breast-ovarian cancer. Most mutations found are of missense type and located in the central region of the gene (exons 5 to 8). In the present study, a germline p53 alteration was identified in a late onset breast cancer family (kindred Lund 5; mean age 58 years) using single stranded conformation polymorphism and sequence analysis. The mutation (a CCG to CTG transition) at codon 82 in exon 4, resulting in a proline to leucine substitution, has not previously been reported and was not present in a control set of 60 healthy individuals. Three of five woman with breast cancer (45, 57 and 65 years) were carriers of the alteration. Loss of heterozygosity at the p53 locus was not seen in the primary tumors of these women, but appeared as a partial loss of the wildtype allele in subsequent recurrent lesions of two gene carriers. The family manifested no linkage to the p53 gene (a two-point LOD-score of -0.41), and has previously also been excluded for linkage to the BRCA1 and BRCA2 loci, as well as being carrier of a BRCA1 germline mutation. Although it seems unlikely that the p53 germline mutation is the major cause of disease predisposition in Lund 5, the data suggest that some p53 alteration may confer a subtle influence on breast cancer development and progression. PMID- 8710379 TI - Tyrosine kinase inhibitor herbimycin A reduces the stability of cyclin-dependent kinase Cdk6 protein in T-cells. AB - The Cdk6 protein level rapidly decreased after treatment with the tyrosine kinase inhibitor herbimycin A in human T-cell lines. This decrease is fairly specific, because the level of other Cdks such as Cdk2 and Cdk4 and cyclins such as cyclin D2, cyclin E and cyclin A, did not change significantly even after 24 h treatment with herbimycin A. Pulse-chase analysis revealed that the decrease in the Cdk6 protein level results from reduced stability of the protein. Our results suggest that herbimycin A-sensitive protein tyrosine kinase(s) are involved in the regulation of the Cdk6 protein level. PMID- 8710381 TI - Discordance between accumulated p53 protein level and its transcriptional activity in response to u.v. radiation. AB - In response to DNA damage, the transcriptional activity of p53 rises. This has been thought to be due to an increase in the level of p53 protein. By comparing the p53 protein level and its ability to transactivate target genes Waf1/Cip1 and mdm2 in both T22 and NIH3T3 cells irradiated with u.v., a discordance between the p53 protein level and its transcriptional activity was observed. When the cells were irradiated with 10 J/m2 of u.v., there was a substantial increase in expression of Waf1/ Cip1 and mdm2. However, little increase in Waf1/Cip1 and mdm2 expression was observed in T22 and NIH3T3 cells 8 or 9 h after exposure to 50 J/m2 of u.v., although the p53 protein level accumulated to its highest level under these conditions. Interestingly, a significant increase in Waf1/Cip1 expression was seen 24 h after irradiation in NIH3T3 cells, indicating that the inhibition of p53 transcriptional activity is reversible. Discordance between the transcriptional activity of p53 and its protein level was further studied using a cell line expressing the p53 reporter plasmid RGC delta fosLacZ. Using double immunofluorescence staining, the coexpression of p53 and beta-galactosidase from the reporter plasmid in the same cells was investigated. The observed lack of correlation between the elevated p53 and beta-galactosidase and expression in u.v. irradiated cells strongly indicates that the ability of p53 to transactivate its target genes is not simply correlated to its protein level. The results indicate that the transcriptional activity of p53 may be negatively regulated. PMID- 8710382 TI - Enhanced protein stability: a novel mechanism of D-type cyclin over-abundance identified in human sarcoma cells. AB - The mammalian D-type cyclins promote progression through a G1 checkpoint by phosphorylating the retinoblastoma protein (pRB), and can contribute to oncogenesis via their deregulated expression achieved through gene amplification, chromosomal rearrangement, or retroviral integration. We now report a novel mechanism of tumour-associated D-cyclin over-abundance, resulting from enhanced protein stability. In two human cell lines established from a single uterine sarcoma biopsy, pRB-positive SK-UT-1B and pRB-deficient SK-UT-1, aberrant accumulation of functional cyclins D1, and D2 and D3 occurred in the absence of gene amplification and/or elevated mRNA expression. The abundance of D-cyclin proteins remained elevated throughout the cell cycle, and pulse-chase experiments revealed six to 10-fold prolongation of their protein half-lives as compared with either diploid fibroblasts or control U-2-OS sarcoma cells. These results point to a critical regulatory role of D-type cyclin turnover, and contribute to refinement of current views of the role played by the cyclin D-CDK-p16-pRB pathway in cell cycle control and tumorigenesis. PMID- 8710383 TI - Expression of HPV16 E6 or E7 increases integration of foreign DNA. AB - In most invasive cervical carcinomas, high-risk human papillomavirus (HPV) DNA is integrated into the host genome, while in pre-invasive cervical lesions the viral genome is typically maintained exclusively as an episome. In contrast, integration of low-risk HPV DNA is rare, as is the association of low-risk HPVs with carcinomas. High-risk HPV integration is associated with a selective growth advantage of affected cells, and hence, integration is likely to be an important genetic alteration contributing to cervical tumor progression. Expression of high risk, but not low-risk, HPV E6 or E7 proteins disrupts the p53-dependent G1 arrest that cells normally display in response to DNA damage. Absence of this cell cycle checkpoint may predispose cells containing high-risk HPVs to genetic instability and to the accumulation of the genetic alterations that appear to be required for HPV-associated cervical tumor progression. We hypothesized that integration of high-risk HPV DNA into the host cell genome may be facilitated by E6- and/or E7-mediated disruption of the normal DNA damage response pathway. To test this hypothesis, we assessed the integration frequency of a reporter plasmid (pHyGal) in RKO cells expressing individual E6 or E7 genes of either high-risk (HPV16) or low-risk (HPV6, HPV11) type viruses. Cells expressing HPV16 E6 or HPV16 E7 exhibited a significantly increased frequency of pHyGal integration in comparison to RKO control cells or cells expressing low-risk HPV E6 or E7. Thus, expression of high-risk, but not low-risk, E6 and E7 proteins increases the frequency of foreign DNA integration into the host genome. These findings suggest that at least some of the difference in oncogenic potential observed between high risk and low-risk HPV types may be determined by the increased ability of high risk HPVs to integrate into host DNA. PMID- 8710384 TI - Telomerase activity in normal human epithelial cells. AB - Telomerase activity is found in most cancer tissues and many immortalized cell lines as well as in germ line cells but it is generally undetected in normal human somatic tissues. There is weak telomerase activity in some cell types of hematopoietic lineage in which a stem cell-like subpopulation may exist. Likewise, physiologically regenerating somatic tissues and organs such as skin, small intestine, and most other epithelia of the human body are supposed to contain similar cell lineages to maintain their renewal throughout the life span of individuals. It is therefore of interest whether telomerase activity is present in physiologically regenerating epithelial cells. Telomerase activity was detected, though very weakly, in cultured normal epidermal keratinocytes and at higher levels in a subpopulation that adhere rapidly on collagen IV-coated culture dishes. No telomerase activity was detected in a subpopulation that was less adherent on the coated dishes. The rapidly adherent subpopulation of keratinocytes was enriched in small proliferating cells with macrocolony forming potential. It was also passaged through more generations in culture, and expressed integrin beta 1 at higher levels than the less adherent subpopulation. Telomerase activity was similarly found in ectocervical keratinocytes as well as in simple endocervical epithelial cells. These findings provide the evidence of a telomerase-positive population among physiologically regenerating normal human epithelial cells. The identity of the telomerase-positive cells remains to be defined. PMID- 8710385 TI - Domains of the qin protein required for oncogenic transformation. AB - The qin oncogene is a cell-derived insert in the genome of avian sarcoma virus 31 (ASV 31) and functions as the oncogenic determinant of that virus. Overexpression of the viral and cellular versions of the Qin protein (v-Qin and c-Qin) induces oncogenic transformation of chicken embryo fibroblasts (CEF); v-Qin also rapidly induces fibrosarcomas in chickens. Qin proteins can bind to specific DNA sequences and act as transcriptional repressors. In this study, mutants of Qin were constructed in order to determine the molecular domains required for transformation of chicken embryo fibroblasts. Our data indicate that three regions required for transforming activity are located (i) between residues 74 141 at the amino terminus, (ii) in the winged helix domain and (iii) between residues 383-395 at the carboxyl terminus. A Qin mutant with 12 amino acids deleted from the carboxyl terminus (383-395) showed transforming activity that was lower than that of wild type Qin for CEF. Compare to wild type Qin transformants, the mutant transformed cells had a reduced ability for multilayered and for anchorage independent growth. Deletion of 48 amino acids from the carboxyl terminus of the Qin protein (347-395) completely abolished transforming activity. In contrast, deletion of 74 amino acids from the amino terminus did not affect transformation of CEF. However, further deletion of 68 amino acids (74-141) reduced but did not abolish transforming activity. Finally, deletion in the winged helix domain (218-295) completely abrogated oncogenic capacity in CEF. These results suggest that DNA binding and transcriptional repression may be important in Qin-induced oncogenic transformation. PMID- 8710386 TI - Testing NF-kappa B1-p50 antibody specificity using knockout mice. AB - Cell extracts from knockout mice can provide definitive proof of antibody specificity. Two NF-kappa B1-p50 antibodies, sc-114 (a commercial antibody) and NR1157, were observed to recognize proteins having distinct electrophoretic mobilities of 52-55 kD and 50 kD, respectively, by Western blot analysis. In order to discriminate the specificity of these antibodies for NF-kappa B1-p50, whole cell extracts derived from NF-kappa B1-p105 knockout mice were employed. While the NR1157 antibody completely failed to recognize its 50 kD product in p105-/- knockout extracts, the sc-114 antibody still strongly recognized its 52 55 kD product. These data demonstrate that NR1157, but not sc-114, is highly specific for NF-kappa B1-p50 by Western blot analysis. In addition, these results highlight the utility of knockout cell extracts for discerning antibody specificity. PMID- 8710387 TI - 100th Annual meeting of the American Academy of Otolaryngology-Head and Neck Surgery Foundation, Inc. Washington, D.C. September 29-October 2, 1996. Abstracts. PMID- 8710388 TI - Good patient care is the bottom line. PMID- 8710389 TI - The antitrust question: a new perspective. PMID- 8710390 TI - How will new risk-sharing policies affect you? PMID- 8710391 TI - Physician-hospital ventures: is there added value? PMID- 8710392 TI - Court affirms relatives' right to shut off life support. PMID- 8710393 TI - Making a difference in Bosnia. Interview by Robert P. Carlson. PMID- 8710394 TI - New therapeutic agents marketed in 1995. PMID- 8710395 TI - Enzyme deficiencies in neonates with jaundice. AB - Numerous findings have shown that enzyme deficiencies, especially those involved in the protection of red cells from oxidation may lead to hemolysis and hyperbilirubinemia. It is established that G6PD deficiency may be the cause of neonatal hyperbilirubinemia, as has been found in several countries and among widely different ethnic groups. We try to establish the incidence of G6PD, PK and GSSG-R deficiencies in neonates with jaundice for a better assessment of the population at risk. The present investigation was carried out in the attempt to be certain whether these enzymes could play a part in the development of neonatal jaundice. A total of 341 neonates of both sexes with jaundice were analyzed: 47 with G6PD deficiency; 9 with PK deficiency and 2 with GSSG-R deficiency. PMID- 8710396 TI - Clinical assessment of the relationship of Helicobacter pylori to gastroduodenal pathologies. A prospective analysis of 253 consecutive patients. AB - The aim of our study was to evaluate the relationship between Helicobacter pylori infection and various gastroduodenal diseases. We also took into consideration alcohol intake and smoking. Two-hundred and fifty-three consecutive patients with ulcer-like symptoms underwent gastroscopic and histological examinations. H. pylori status was evaluated by means of culture, Giemsa stain and CP-test, upon obtaining bioptic samples of gastric mucosa. Two hundred eighteen patients were affected by gastritis, 171 of which were H. pylori positive (78.4%); of 164 patients with active gastritis, 158 were H. pylori positive (96.3%). In 63 patients with duodenal ulcer, H. pylori was present in 58 cases (92.1%). Of 14 patients with gastric ulcer, 11 were H. pylori positive (78.6%). Out of 133 patients with duodenitis associated with active ulcer or a history of previous ulcer, H. pylori was found in 112 patients (84.2%). Among the 27 patients with "autonomous" duodenitis, 18 were H. pylori positive (66.6%); the comparison between the two groups of patients with duodenitis concerning H. pylori infection was statistically significant (p = 0.033). Of the 119 patients tested for acquired MALT, 39 were found positive (32.7%); among these 34 patients were H. pylori positive (87.1%) and only 5 patients were H. pylori negative (12.9%). Thus our study confirms the importance of H. pylori in gastroduodenal pathology underlining its role in the development of acquired MALT (mucosa associated lymphoid tissue) for its possible evolution in low grade B cell primary gastric lymphoma. Alcohol intake and smoking do not appear to play a role in H. pylori infection. PMID- 8710397 TI - Serial in vitro transfer of tuberculin hypersensitivity by sensitised lymphocytes. AB - An account is given of the experimental serial in vitro transfer of PPD-specific delayed hypersensitivity to peripheral leukocytes using PPD-specific TF solely as the initial source. Transfer was assessed by means of the leukocytes migration inhibition test, the lymphocytes locomotion test and the leukocytes adherence inhibition test. These tests were positive in all the experiments. It is suggested that these results indicates that TF does not act in itself but triggers a reaction that expands the effect of hypersensitivity. PMID- 8710398 TI - Serial in vitro transfer of hypersensitivity to cancer antigens by sensitised lymphocytes. AB - An account is given of the experimental serial in vitro transfer of antigen specific delayed hypersensitivity to peripheral leukocytes, using antigen specific Transfer factor solely as the initial source. Transfer was assessed with the leukocyte migration, lymphocyte locomotion and leukocyte adherence inhibition tests. The positive test results observed in all the experiments suggest that Transfer factor does not act as such, but triggers a reaction that expands the effect of hypersensitivity. PMID- 8710399 TI - Alternative saphenous vein sparing surgery for future grafting. AB - OBJECTIVE: Evaluation of long saphenous vein sparing surgical procedures alternative to high ligation and distal stab avulsion, in terms of effectiveness and suitability for eventual by-pass surgery. EXPERIMENTAL DESIGN: Prospective evaluation of 125 operations for primary varicose veins, 52 external valve plasties of the sapheno-femoral junction (EV-SFJ) (42 performed using the hand sewing technique and 10 using the Veno-cuff device), mean follow-up 45 months, and 73 hemodynamic correction of varicose veins (French acronyms: CHIVA), mean follow-up 30 months. SETTING: Department of General Surgery, University of Ferrara. Institutional practice, one-day surgery. PATIENTS: Patients were selected using clinical, Doppler cw, and duplex scanning evaluations. Patients with early varices due to sapheno-femoral reflux with duplex scanning evidence of mobile valve leaflets underwent EV-SFJ. The other patients were operated on using the hemodynamic correction technique. Both groups underwent preoperative ambulatory venous pressure (AVP) and light reflection rheography-refilling time (LRR-RT) measurements. INTERVENTIONS: EV-SFJ restores valve function correcting vein wall dilitation by applying an external prosthesis. CHIVA consists of selected ligatures of the superficial veins that allow superficial blood aspiration in the deep veins through the perforators. MEASURES: The outcome was evaluated with clinical and ultrasonographic examinations, AVP and LRR-RT measurements. RESULTS: Long saphenous vein patency registered after EV-SFJ and CHIVA was 94.2% and 90.4%, respectively. Both treatments preserve the drainage function in the saphenous system. Varicose veins recurrence percentage rate was 9.6% and 10.9%, respectively. CONCLUSIONS: Following the proposed selection criteria, these two alternative procedures seem to be more effective in varices treatment than high ligation and have the advantage of preserving saphenous veins suitable for eventual by-pass surgery. PMID- 8710400 TI - Mitral valve repair using continuous warm blood cardioplegia. AB - To achieve satisfactory mitral valve repair, we applied continuous warm blood cardioplegia to seven children with congenital heart disease associated with mitral valve insufficiency. All patients had mild to moderate mitral valve regurgitation before the operation. The competency of the mitral valve was assessed by injecting test solution into the left ventricle through the mitral valve orifice with the heart beating under warm blood perfusion into the aortic root proximal to the cross-clamp. Repair of the mitral valve was performed under normothermic cardiac arrest with continuous antegrade perfusion of warm blood cardioplegia. In all patients, these two procedures could be alternated with ease and safety simply by varying the potassium concentration of cardioplegia. While satisfactory valve repair was obtained with the initial procedure in 4 cases, repair was repeated twice in two and three times in one. There were no operative deaths and no complications directly related to the procedure. Postoperative echocardiography demonstrated good mitral valve function with none to trivial regurgitation in six patients (86%). Continuous warm blood cardioplegia is a useful and safe tool for mitral valve repair. PMID- 8710401 TI - Repair of femoral pseudoaneurysm after cardiac catheterization: utility of extraperitoneal iliac control. AB - Femoral artery lesions may occur after cardiac catheterization or percutaneous transluminal coronary angioplasty (PTCA). The incidence of pseudoaneurysm following transfemoral catheterization ranges from 0.21 to 6.25%. Among 3162 cardiac catheterization procedures thirteen (0.41%) patients presented a femoral pseudoaneurysm requiring surgical repair. The Authors reported their technical consideration about the utility of extraperitoneal iliac control during surgical repair of femoral artery pseudoaneurysm due to cardiac catheterization. PMID- 8710402 TI - Indications and role of isolated profundaplasty in patients affected by limb ischemia. AB - The deep femoral artery is the primary source of blood supply to the lower extremity in presence of superficial and/or popliteal occlusion. The atherosclerotic involvement of the profunda femoris artery is relatively less frequent and generally is localized in the initial segment of the artery. The profundaplasty employed to relieve limb threatening ischemia is infrequently used as an isolated procedure. However many authors demonstrated that restoration of flow through this vessel alone, in patients without significative lesions of the aorto-femoral district, will effectively relieve ischemia when the superficial femoral artery is also occluded. Our experience gained in the last eight years concerns 24 patients (21 male, 3 female). All the patients had severe ischemia of the lower limb with invalidating claudication (13), rest pain (9) and gangrene (2). On the basis of our results, we think that the operation is recommended, whenever possible, in patients with critical ischemia when the possibilities of more extensive revascularizing procedures are absent. PMID- 8710403 TI - One surgical experience in regime of day hospital: considerations on the first one-hundred patients treated. AB - The development of surgery in regime of day hospital proceeds swiftly, especially in Anglo-saxon countries, so that at the beginning of the second millennium it can be foreseen that in USA alone, 75% of all surgery will be carried out in this manner. From March 1st to September 1st 1994, 100 patients were submitted to operations in ODS (One Day Surgery). We had 3 reconversions into ordinary hospitalization (3%), 2 for social-economic reasons and one for headache and vomiting due to intolerance to local anesthetics. As has been seen we have encountered no important complications, all patients were satisfied. From the analysis of our experience we have deducted useful indications that oblige us to partially modify our attitude: we want to transform our service into a free standing center where the patient can undergo preoperative exams, anesthesiologic examinations and surgery on the same day; we are just about to verify the possibility, thanks to an accurate anamnesis, to not request preoperative routine exams in patients with ASA 1 and 2 physical status; to look for a possible asymptomatic crural hernia in patients that undergo inguinal hernioplasty; we do not submit patients to ODS if they do not have assistance at home; or if they live too far from our service. PMID- 8710404 TI - Effectiveness of nedocromil sodium in preventing exercise-induced asthma in children. AB - Exercise-induced asthma is a well-known phenomenon, particularly affecting children, with an important social impact. In order to assess the usefulness of nedocromil sodium in the prevention of exercise-induced asthma, we studied 49 (15 females, 34 males) children who suffered from asthma; their mean +/- SD age was 9.2 +/- 3.0 (range: 3.3-19.1) years. On the first day respiratory function was evaluated by spirometry, basally and after 6 minutes. The inhalation of nedocromil sodium had a great influence on post-exercise lung function measurements; in fact on the day of nedocromil sodium pretreatment, our patients showed an increase of respiratory function, significantly different from the parameters recorded during the first day. Our findings suggest that nedocromil sodium is effective in the prevention of exercise-induced asthma, in pediatric age. PMID- 8710405 TI - Selective treatment and targeted chemotherapy: effect on prevalence and intensity of infection for two intestinal helminths in Nepalese children. AB - The study examines the effect of mebendazole distribution (2 x 100 mg x 3 days) either as mass chemotherapy targeted to children 1 to 10 years old, or selective treatment of heavily infected children, over a two year period in sixteen rural villages of Nepal. Eight villages were covered with six monthly mass chemotherapy, provided by Community Health Volunteers, other eight with yearly selective treatment of heavily infected children based on clinical signs. Prevalence and intensity of infection, indirectly measured as the number of eggs per gram of faeces, were measured for A. lumbricoides and hookworms utilising the Kato thick-smear technique. At the end of two year intervention, the effect of both activities was similar on worm prevalence with a slightly significant difference (p < 0.05) on worm load in favour of mass chemotherapy. Furthermore a greater effect of mebendazole was reported, regardless of the type of intervention, on hookworms present at baseline with a lower prevalence (23.4%) than A. lumbricoides (47.6%). The study suggests that clinical diagnosis of heavily infected children may be a feasible way to direct selective treatment. Baseline prevalence seems to influence effectiveness of control programs. Further studies are needed to substantiate these observations. PMID- 8710406 TI - Changes in plasma volume during prolonged restriction of motor activity: effect of chronic hyperhydration. AB - Prolonged reduction of number of steps taken per day by healthy subjects (hypokinesia), is associated with a decrease in plasma volume (PV), which may contribute to the development of several so-called hypokinetic disorders seen immediately during exposure to hypokinesia (HK). The purpose of this study was to determine whether a daily intake of fluid and salt supplementation (FSS), would attenuate this loss of PV. Thirty male long distance runners (20-25 years of age) completed this investigation. Ten volunteers placed under a normal ambulatory life (control subjects), ten volunteers were subjected to continuous restriction of motor activity without using FSS (hypokinetic subjects) and ten volunteers were submitted to continuous restriction of motor activity and took daily FSS (hyperhydrated subjects). For the simulation of the hypokinetic effect the hypokinetic and hyperhydrated volunteers were kept under 2.7 km/day (3,000 steps/day). Plasma volume decreased significantly (p < 0.05) in the hypokinetic volunteers during HK, from 3.010 +/- 81 to 2.693 +/- 52 ml. The volunteers who were submitted to combined HK and FSS, had a significant (p < 0.05) increase in PV during HK, from 2.940 +/- 72 to 3.327 +/- 72 ml. There were demonstrable significant differences in PV between the hyperhydrated and hypokinetic groups of volunteers. Thus, prolonged restriction of motor activity appears to have significant effects on the loss of PV, whereas chronic hyperhydration significantly increases PV. PMID- 8710407 TI - Phytosterols as growth regulators. AB - Starting from squalen and gonan structures, nature produces by enzymatic transformation a great variety of P--and subsequently also of "animal" sterols. This wonderful chain of biosyntheses comprises a number of provitamins such as beta-carotene and ergosterol and has an endpoint in the vitamin D compounds ergocalciferol (ercalciol, vit D2) and cholecalciferol (calciol, vit D3). On the other hand, we have has an endpoint the most important compound cholesterol and its derivatives, like androsterone, testosterone and 17 beta-estradiol. The members of this family of chemical compounds perform the most diverse biological functions, if properly "directed" or transported to their respective sites of action. Since all of these compounds are hardly soluble in water, we concentrated our research work on their solubilization and thus on enhancing their bioavailability and bioreactivity. The bioreactivity of P was taken as guiding principle for an investigation into understanding the ways of nature as closely as possible-and, at same time-to interpret the findings in comprehensive context. An effort at integrated system's thinking was made throughout. PMID- 8710408 TI - New pathogens, and diseases old and new. I) Afipia felis and Rochalimaea. II) Parvovirus B 19. III) herpesvirus 6. AB - The paper describes events that in the last fifteen years, have led to the identification of the aetiological agents of three widely known diseases: cat scratch disease, erythema infectiosum and exanthem subitum. The particular features of Afipia felis and Rochalimaea, Parvovirus B 19 and Herpesvirus 6 are presented. The paternity of new diseases (i.e. bacillary angiomatosis, bacillary peliosis hepatitis, LES-like syndrome, chronic fatigue syndrome, petechial glove and sock syndrome, etc.) has also been attributed to some of these pathogens as has the paternity of some older ones (i.e. aplastic crisis, erythroblastosis fetalis, trench fever, hepatitis, opportunistic infection, etc.). It has been argued that the same pathogen can cause different diseases depending on the immunogenic state of the subject. To date, persisting difficulties in isolating the pathogen or differentiating between latent or active infection, still in some cases raises doubts concerning the attribution of the disease to a specific agent. New immunological or molecular techniques, allowing the direct detection of in vivo replication, are still needed in order to establish a sure connection between some of these agents and some of these diseases. Progress here will both give more accurate data about the epidemiology of some diseases and allow us to apply more appropriate treatment and prevention techniques. PMID- 8710409 TI - Pre-B acute lymphoblastic leukemia in a patient with partial lipodystrophy and acanthosis nigricans. AB - In patients with lipodystrophies a post binding defect in insulin action has been described involving phosphorylation of the beta subunit of the insulin receptor, suggesting the presence of a genetically determined defect in insulin action; the receptor gene has been mapped to the distal short arm of chromosome 19 close to the break-point of a specific chromosome translocation frequently found in pre-B Acute Lymphoblastic Leukemia (ALL). We report on a 13 years old female patient with partial lipodystrophy, acanthosis nigricans and insulin resistance who developed a pre-B ALL. Since lipodystrophy and pre-B ALL are rare disorders, a possible causal relationship between the two diseases is suggested possibly mediated by a mutation in the insulin receptor gene. PMID- 8710410 TI - The capacity of the fungus Duddingtonia flagrans to prevent strongyle infections in foals on pasture. AB - A field trial was conducted to evaluate the potential of the nematode-destroying fungus Duddingtonia flagrans to control free-living stages of horse strongyles. In late Spring 2 groups of horses (yearlings) with mixed infections of strongyles were allowed to contaminate 2 equal-sized pastures. One of the groups (F) received a daily dose of D. flagrans mixed in a feed supplement, while the other (C) received a similar amount of supplement without fungus. During a 3-month contamination period strongyle egg counts in faeces and number of infective strongyle larvae harvested from faecal cultures were determined. Grass samples were collected fortnightly. After the contamination period the yearlings were removed and 2 groups of young tracer foals (TF and TC) grazed the fungus and control pastures respectively for 4 weeks, housed for another 15 weeks and then killed to determine their worm burdens. The number of larvae in cultures from group TF was significantly lower than that in TC and herbage infectivity was reduced to a very low level on the pasture grazed by horses fed fungi. The number of Strongylus vulgaris and Strongylus edentatus larvae was also significantly lowered in group TF. Cyathostome larvae recovered from the mucosa of the ventral and dorsal colon and from the caecum were significantly lowered in group TF foals. Also, the number of strongyles found in the gut contents of group TF foals were significantly reduced in the dorsal colon, but numbers of worms in the ventral colon and in the caecum were similar to those of the controls. PMID- 8710411 TI - Hypoxanthine: a low molecular weight factor essential for growth of erythrocytic Plasmodium falciparum in a serum-free medium. AB - A low molecular weight factor in a basal medium essential for erythrocytic Plasmodium falciparum development in a serum-free medium using a cell growth promoting factor derived from adult bovine serum was detected. The factor was hypoxanthine. The optimal hypoxanthine concentration for parasite growth was between 15 and 120 microM. The contribution of hypoxanthine to increased parasite growth was clearly evident in cultures on day 4. Among various low molecular weight supplements tested, adenine, adenosine, AMP, ATP, cyclic AMP, guanine, guanosine, inosine, inosine monophosphate, xanthine, NAD, NADH, NADP, NADPH and deoxyguanosine triphosphate showed a similar effect to that of hypoxanthine in the serum-free culture system. On the other hand, the addition of uric acid, FAD, thymidine, uridine, orotic acid, deoxythymidine triphosphate, deoxycytidine triphosphate, deoxyadenosine triphosphate, ribose-1-phosphate, or ethanolamine was not beneficial to the parasite growth. The results presented here will not only be of practical value, but will provide important information about the developmental requirements of the parasite. PMID- 8710412 TI - The within-host cellular dynamics of bloodstage malaria: theoretical and experimental studies. AB - The properties of a mathematical model of bloodstage infection with a single strain of malaria were investigated. Analysing the cell population dynamics in the absence of a host immune response we demonstrate a relationship between host and parasite parameters that defines a criterion for the successful invasion and persistence of the parasite. Important parameters are the rates of merozoite production and death and those of erythrocyte production, death and invasion. We present data from experiments designed to evaluate the erythrocyte invasion rate in a rodent malaria system. The model generates patterns of parasitaemia in good qualitative agreement with those seen in Plasmodium berghei infections. The sole force behind the rise and fall in parasitaemia in the model without immunity is the density of susceptible erythrocytes, suggesting that resource availability is an important determinant of the initial pattern of infection in vivo. When we incorporate a simple immune response into the model we find that immunity against the infected cell is much more effective at suppressing parasite abundance than immunity against the merozoite. Simulations reveal oscillating temporal patterns of parasite abundance similar to P. c. chabaudi infection, challenging the concept that antigenic variation is the sole mechanism behind recrudescing patterns of infection. PMID- 8710413 TI - Enzyme polymorphism in Endotrypanum and numerical analysis of isoenzyme data. AB - In this study, we have analysed enzyme polymorphism among a group of protozoan parasites of the genus Endotrypanum (Kinetoplastida: Trypanosomatidae). Seventeen stocks of Endotrypanum spp. isolated from sloths (Choloepus didactylus and C. juruanus) in the Amazon Region of Brazil were analysed by enzyme electrophoresis, and their electromorphic profiles were compared with reference strains reported previously. The 16 enzymic loci were analysed, and the strains were classified into zymodemes, each representing parasites with unique enzyme profiles. Each zymodeme was considered as an elementary taxon, and using numerical analyses (cladistic, agglomerative hierarchical and ordination techniques) the genus was shown to be monophyletic and the 12 zymodemes characterized could be divided into 3 groups (A, B, C). The heterogeneous population (which may represent a complex of parasite species or strains variants) showed, however, no correlation with the origin (i.e. host species involved or geographic area of isolation) of Endotrypanum stocks. Eight isolates of Endotrypanum sp. from Rondonia State (Brazil) and a parasite strain from Panama were clustered together into a zymodeme, which was phenetically closely related to the E. monterogeii from Costa Rica. The data indicate that E. schaudinni is a species complex. PMID- 8710414 TI - Induction of changes in human behaviour by the parasitic protozoan Toxoplasma gondii. AB - Toxoplasma gondii, the coccidian parasite, is known to induce changes in the behaviour of its intermediate hosts. The high prevalence of this parasite in the human population (20-80%) offers the opportunity of studying the influence of the parasite on human behaviour by screening of a normal population. A total of 224 men and 170 women were tested for toxoplasmosis and their personality profiles were measured by Cattell's questionnaire. Highly significant differences between Toxoplasma-infected and uninfected subjects were observed (P < 0.01). For men the factors G (low superego strength), L (protension), O (guilt proneness), and Q2 (group dependency) were positively influenced in infected subjects. For women the prevailing factors were A (affectothymia, P < 0.01), L (alaxia), O (untroubled adequacy) and Q2 (self-sufficiency). To reveal whether toxoplasmosis induces personality factor-shifts or whether certain combinations of personality factors influence the probability of acquiring Toxoplasma infection, we examined the personality profiles of 164 male patients diagnosed with acute toxoplasmosis during the past 13 years. The existence of a positive correlation between the duration of latent toxoplasmosis and the intensity of superego strength decrease (P < 0.02) suggested that the decrease of superego strength (the willingness to accept group moral standards) was induced by T. gondii infection. PMID- 8710415 TI - Cholinergic inhibition of muscle fibres isolated from Schistosoma mansoni (Trematoda:Digenea). AB - Cholinergic compounds inhibit FMRFamide-induced contractions in dispersed muscle fibres isolated from adult Schistosoma mansoni. Acetylcholine (ACh) was the most effective cholinergic agonist tested with an EC50 < 100 nM. Less effective were propionylcholine and arecoline with EC50 < 1 microM and butyrylcholine and carbachol with EC50 < 10 microM. Choline, muscarine, pilocarpine, nicotine, DMPP (1,1-dimethylphenylpiperazine) and levamisole were all ineffective. Amongst tested antagonists, d-tubocurarine (100 microM), mecamylamine (1 mM), scopolamine (1 mM) and quinuclidinyl benzilate (10 microM) were all ineffective. Bicuculline, picrotoxin and strychnine were also ineffective. However alpha-bungarotoxin, at 100 nM, was able to block the inhibitory ACh effect. From these data it appears that the cholinergic receptor on the schistosome muscle fibres may be of the nicotinic type, but that its pharmacology is different from that of nicotinic receptors of vertebrates as well as of nematodes or a variety of other invertebrates. PMID- 8710416 TI - Isolation, characterization and immunolocalization of a globin-like antigen from Ostertagia ostertagi adults. AB - Western blot analysis using an anti-globin rabbit serum Rb94 revealed a major band of 17 kDa in extracts of Ostertagia ostertagi adults and 4th-stage larvae. The adult stage globin-like antigen (OoAdGlb) was purified from total worm extracts by liquid chromatography. The protein has an estimated molecular mass of 36 kDa under non-reducing conditions, suggesting a dimeric structure containing 2 non-covalently linked 17 kDa monomers. Tryptic peptides were sequenced and showed strong similarities with the globins of free-living and parasitic nematodes. Immunolocalization revealed the presence of this globin-like antigen in the body wall musculature and/or the cuticle of O. ostertagi adults. An enzyme-linked immunosorbent assay based on the purified OoAdGlb showed no differences in response between calves infected by O. ostertagi and/or Cooperia oncophora and the negative controls. PMID- 8710418 TI - The ultrastructure of the anterior end of male Onchocerca volvulus: papillae, amphids, nerve ring and first indication of an excretory system in the adult filarial worm. AB - A detailed morphological investigation of the anterior sensory organs, the nerve ring and a glomerulus-like structure in male Onchocerca volvulus was performed by means of electron microscopy. The 8 head papillae are arranged in the common 4 + 4 pattern of most filarial worms in circles around the mouth opening. The amphidial openings are found between the circles of inner and outer papillae on both sides of the mouth. Inside, several additional nerve axons are seen in the tissue of the anterior tip not related to one of the identified papillar structures. The inner and outer papillae exhibit a remarkably different fine structure, and are part of a complex system of at least 2 different receptor cell types at the anterior tip of the worm. The amphidial channel contains 8 modified cilia; accessory axons are associated with the cytoplasm of the sheath cell. The anterior nerve ring of male worms is located about 150 micrometers posterior from the outermost tip of the head region. It consists of several fibres coiled around the oesophagus. The comparison of the fine structure of the central nervous system did not show the expected morphological differences associated with the heterogeneous age distribution in the natural worm population. This was in contrast to previous findings with respect to tissues in different parts of the worm. The study also provides the first evidence that suggests the existence of an excretory organ in a filarial worm in the region of the anterior nerve ring. Paired glomerulus-like structures in the lateral chords and a canal formed by a projection of the basal zone of the cuticles were identified. PMID- 8710417 TI - Resistance of glucose-6-phosphate dehydrogenase deficiency to malaria: effects of fava bean hydroxypyrimidine glucosides on Plasmodium falciparum growth in culture and on the phagocytosis of infected cells. AB - The balanced polymorphism of glucose-6-phosphate dehydrogenase deficiency (G6PD-) is believed to have evolved through the selective pressure of malarial combined with consumption of fava beans. The implicated fava bean constituents are the hydroxypyrimidine glucosides vicine and convicine, which upon hydrolysis of their beta-O-glucosidic bond, became protein pro-oxidants. In this work we show that the glucosides inhibit the growth of Plasmodium falciparum, increase the hexose monophosphate shunt activity and the phagocytosis of malaria-infected erythrocytes. These activities are exacerbated in the presence of beta glucosidase, implicating their pro-oxidant aglycones in the toxic effect, and are more pronounced in infected G6PD- erythrocytes. These results suggest that G6PD- infected erythrocytes are more susceptible to phagocytic cells, and that fava bean pro-oxidants are more efficiently suppressing parasite propagation in G6PD- erythrocytes, either by directly affecting parasite growth, or by means of enhanced phagocytic elimination of infected cells. The present findings could account for the relative resistance of G6PD- bearers to falciparum malaria, and establish a link between dietary habits and malaria in the selection of the G6PD- genotype. PMID- 8710419 TI - Targeted anthelminthic treatment of school children: effect of frequency of application on the intensity of Ascaris lumbricoides infection in children from rural Nigerian villages. AB - A study to compare the effects of different frequencies of targeted chemotherapy with levamisole (Ketrax: JAGAL Pharma, Lagos, Zeneca Pharmaceuticals UK) as an action for the control of Ascaris lumbricoides was carried out in 3 communities in rural Oyo State, Nigeria. The targeted group comprised children, age 5 to 15 years, attending primary school within their village. Treatment was provided within the school with the assistance of the school teachers. Three frequencies of targeted treatment were offered. In one village targeted treatment was provided on 1 occasion in 1 year, in another village at two 6-monthly intervals and in the third village every 4 months. Prevalence and intensity (e.p.g.) of A. lumbricoides infection were determined immediately before and after the period of intervention using a modified Kato-Katz technique. In the villages which received treatment once and at 6-monthly intervals, a reduction in post-treatment intensity of A. lumbricoides was observed in the total population but this failed to attain statistical significance. In contrast, within the village which received 4-monthly targeted chemotherapy, a significant reduction in post treatment intensity of A. lumbricoides was observed in the total population and in the targeted children. In general, reductions in the intensity of A lumbricoides after intervention were not particularly pronounced in untreated children (aged 0-4 years) even in the 4-monthly targeted village, whereas in untreated adults, reductions approached statistical significance in villages which received targeted treatment once and at 4-monthly intervals. PMID- 8710420 TI - [Physiological and pathophysiological role of histamine in the gastrointestinal tract]. AB - In this paper, the authors discuss problems concerning the role played by histamine in various physiological and pathological processes in the gastrointestinal tract. The synthesis and metabolism of histamine is described. Apart from participation in many physiological processes, e.g. secretion of gastric juice, histamine takes part in many pathological reactions occurring in celiac disease, food allergy and inflammatory bowel diseases. PMID- 8710421 TI - [Morphological, microstereological and immunohistoenzymological studies of the jejuni of infants with recurrent constructive bronchitis and chronic diarrhea due to cow's milk protein allergy]. AB - Morphological, microstereological and immunohistoenzymatic technique were used to assess jejunal biopsy samples obtained from 17 infants suffering from chronic diarrhoea and bronchitis spastica (the study group), and 5 infants with chronic diarrhoea and weight deficiency (group G). In the first group, histopathological studies revealed a small degree of villous atrophy (II degree) in 12/17 of cases, and in 5/17, a moderate atrophy (III degree). The microstereological technique disclosed infiltration of the submucosa which was significantly more marked in the study group as compared with group G (p < 0.05). Positive immunohistoenzymatic reactions in mononuclear cells of the submucosa infiltration were observed in all cases, but was more distinct in the first group. The results of the study show jejunal biopsy to be helpful in confirming the allergic etiology of the disease in the study group. PMID- 8710422 TI - [The influence of the feeding type on the concentration of trace elements in the hair of infants]. AB - The aim of this study was to assess hair magnesium, iron, copper, and zinc levels in breast-fed and formula-fed infants. Our findings showed that concentration of the assayed macro- and trace elements, except copper, were higher in breast-fed children and declined with age, faster in formula-fed children. The frequent occurrence of magnesium deficiency indicates that pregnant women should receive magnesium supplementation. PMID- 8710423 TI - [Arterial hypertension in children aged 8-14 living in Zabrze]. AB - In a group of 465 children (232 girls and 233 boys) aged 8-14 years, attending a primary school in Zabrze (Katowice District, southern Poland) arterial blood pressure was measured 3 times, body height and weight, once. A questionnaire collecting information about parents' education and hypertension, school performance and physical activity of the children was used. Among the studied children 417 (89.7%) presented with normal mean arterial pressure values, while 36 (7.7%) with arterial hypertension and 12 (2.6%) with borderline hypertension. The influence of children's obesity and parents' hypertension on the incidence of hypertension in children was proved, while physical activity, children's school performance as well as education of the parents did not show significant influence. The studied group revealed the highest frequency of arterial hypertension in Poland. Arterial blood pressure measurement is an important element of pediatric examination and treatment, constituting prophylaxis of cardiovascular morbidity in adulthood. PMID- 8710424 TI - [Circadian rhythm of blood pressure in adolescents with hypertension and insulin dependent diabetes mellitus]. AB - Assessment of blood pressure values and their circadian rhythm were performed a group of adolescents with borderline hypertension and in patients with insulin dependent diabetes mellitus. Higher values of blood pressure in children with IDDM in comparison with healthy children and slighter decrease of nocturnal blood pressure in the investigated groups of adolescents were shown. PMID- 8710425 TI - [The influence of physical effort of variable intensity on glycemia in children with diabetes]. AB - The influence of physical effort of variable intensity on glycaemia in children was analyzed in 61 children (29 girls and 32 boys) from 5-17 years of age with newly diagnosed diabetes and with diabetes lasting from 1-12 years (5.5 on average) was assessed. The children underwent an exercise test on a thread null. The pulse rate was on indicator of effort. Pulse rate before and after the effort was measured using a Sport-Tester apparatus and the level of glucose in the capillary bloodstream was determined using an Ames glucometer. Three tests of increasing intensity were performed, each lasting 20 minutes on three consecutive days. In children with well controlled insulin dependent diabetes with glycaemia up to 150 mg/100 ml in first three hours after test, physical effort of variable intensity caused an individually variable fall in glycaemia from 15-43 mg/100 ml. In children with poorly controlled insulin dependent diabetes with hyperglycaemia from 210 to 300 mg/100 ml, physical effort caused a rise of glycaemia from 20-80 mg/100 ml compared to controls. Up to 3 hours after physical effort, 20 children had hypoglycaemia 60 mg 100 ml and 4 children had hypoglycaemia below 40 mg/100 ml. Hypoglykaemia manifested as a mild complex of clinical signs. PMID- 8710426 TI - [Efficacy and safety of dexfenfluramine treatment in obese adolescents]. AB - To assess the effect of dexfenfluramine on weight loss in obese adolescents, a group of 19 obese girls and boys was treated with a low calorie diet, supplemented with dexfenfluramine. The control group comprising the same number of patients was treated with diet only. Three weeks of hospitalization were followed by three weeks of treatment on an outpatient basis. An insignificantly larger weight loss in the dexfenfluramine-treated group was noted during the outpatient treatment. Dexfenfluramine therapy also resulted in a significant reduction of the serum triglycerides level. No serious side effects of dexfenfluramine therapy were noted. PMID- 8710427 TI - [Thrombo-embolic disease as a complication of nephrotic syndrome in children]. AB - Eight children with thrombo-embolic disease in the course of nephrotic syndrome were treated at II Clinic of Children's Diseases (Institute of Pediatrics, Poznan) between 1991 and 1993. The diagnosis was established on the basis of clinical examination and noninvasive imaging techniques. Two patients had an atypical localisation of the thrombus in the left ventricle and right atrium. In laboratory tests of the coagulation system, all of the children had decreased levels of antithrombin III (AT III). All children were treated with heparin and 4 with fibrinolytic agents. AT III concentrate was administered to 3 children. Total resolution of thrombo-embolic disease was obtained in 5 patients, 3 died during treatment. Thrombo-embolic disease should be taken into account in the differential diagnosis of complications of nephrotic syndrome. PMID- 8710428 TI - [One center's experience with fibrinolytic treatment in children]. AB - Fifteen children treated with fibrinolytic agents are presented. The most frequent indication was thromboembolic disease (TED). Eleven patients received streptokinase, 5-urokinase and 3-tissue plasminogen activator. Concomitant heparin was administered to 9 patients with TED. Total resolution was achieved in 9 children, partial improvement in 5; 1 child died during treatment without any improvement. Bleeding complications were observed in 6 patients, 1 of them died due to haemorrhagic stroke. According to the literature and our own experience, we recommend fibrinolytic agents as the treatment of choice for severe TED also in children. PMID- 8710429 TI - [Enterohaemorrhagic Escherichia coli O157:H7 infection]. AB - Escherichia coli O157:H7 differs from previously described diarrheagenic E. coli classes (enteropathogenic, enteroinvasive, enterotoxigenic) by distinct clinical symptoms, production of verotoxin (VT) and a specific plasmid. Cattle are the primary reservoirs of E. coli O157:H7. The organism may be transmitted through the consumption of contaminated foods (mainly of bovine origin) and by person-to person contact. The most typical clinical manifestations of E. coli O157:H7 infection are hemorrhagic colitis and hemolytic-uremic syndrome. Since the 1982 many outbreaks of E. coli O157:H7 infections as well as sporadic cases have been documented. Diagnosis of E. coli O157:H7 is based on a positive stool culture, presence of VT and elevated serum antibodies. The best currently available and inexpensive method for diagnosing E. coli O157:H7 is culture of stool on sorbitol Mac Conkey agar medium. PMID- 8710430 TI - [Thermogenesis during parenteral nutrition and after oral meal]. AB - The aim of the study was to assess thermogenesis during parenteral feeding (group A) and after an oral test meal of a polymeric diet (group B). Carbohydrates, fat and protein ratios in the oral meal and parenteral mixture were: 51:34:15 and 76:11:13, resp. In both situations 20% of resting energy expenditure (REE) was administered-as a bolus oral meal or during 120 min. of intravenous infusion. Mean results of respiratory gas exchange of 15 min. periods were used for calculations. Basal respiratory quotient (RQ) in group A and B was 0.841 and 0.806 resp (difference not significant; NS); peak RQ was 0.910 and 0.924 resp. (NS). Peak RQ in group B was significantly higher than basal (p < 0.006). REE in group A and B was 1,416 +/- 0,231 kj/kg/15 min. and 1,322 +/- 0,226 kj/kg/15 min. resp. (NS). Peak thermic effect (expressed as the rise of REE in percent of the energy content of the nutrients) was: 4.778% and 5.135% resp. (NS). Postprandial thermogenesis is not dependent on the route of administration of nutrients (parenteral or oral). The proportion of substrate utilisation depends on the content of the meal or parenteral mixture. PMID- 8710431 TI - [Recombinant tissue plasminogen activator for thrombolytic therapy of right atrial thrombus in a neonate and infant]. PMID- 8710432 TI - [Acute gastrointestinal hemorrhage in a 12-month-old child following treatment with panadol]. AB - After the treatment of a twelve-month-old child with a small dose of Panadol, we observed an acute gastro-intestinal hemorrhage which required aggressive and ongoing medical intervention. This event should be taken into consideration when antipyretic agents are given to infants and small children. PMID- 8710433 TI - [Anorexia as the main symptom of gastroesophageal reflux: case study of a 13.5 year-old girl]. AB - 13.5-year-old girl who was admitted to hospital because of anorexia nervosa started to complain of abdominal pain in her 3-rd week of hospitalization. She underwent 24-hour pH-metry and upper GI endoscopy. These procedures showed that GER was the cause of anorexia. Therapeutic management led to marked improvement. PMID- 8710434 TI - [Vasovagal syncope in 15-year-old girl]. AB - The authors present a case of a 15-year-old girl with syncope and bradycardia, in whom vasovagal syncope was recognized. The diagnosis was confirmed by positive tilt test. Because of the dominant vasodepressor reaction, Effortil was started to prevent further syncope, with good results. PMID- 8710435 TI - Floating ideas. PMID- 8710436 TI - Multiple completions primed by occlusion patterns. AB - There is a strong tendency to complete a partly occluded shape. Two types of pattern completion, global and local, are frequently reported. By means of the primed-matching paradigm, it has previously been shown that global completions are prevalent for stimuli in which regularity is abundantly present. In our study the primed-matching paradigm is applied to such stimuli in order to find out whether the rival local completion is generated as well. Therefore anomalous completions are added to the experimental design. Priming effects both on global and on local completions are compared with priming effects on those anomalous completions. The data indeed suggest that the occlusion patterns evoked not only a global but also a local completion. PMID- 8710437 TI - Size invariance in visual object priming of gray-scale images. AB - The strength of visual priming of briefly presented gray-scale pictures of real world objects, measured by reaction times and errors in naming, was independent of whether the primed picture of the object was presented in the same size as or different size from the original picture. These findings replicate results on size invariance in shape recognition, which were obtained with line drawings, and extend them to the domain of gray-level images. Entry-level shape identification is based predominantly on scale-invariant representations incorporating orientation and depth discontinuities which are well captured by line drawings. PMID- 8710438 TI - Perceptual latency and complex random-dot stereograms. AB - Naive observers of random-dot stereograms depicting complex surfaces often find that they require several tens of seconds before the impression of depth emerges. With practice, however, perception times often decrease markedly: perceptual learning occurs. Current explanations of these effects were assessed in two experiments. In the first experiment the perception times of naive observers for random-dot stereograms which depicted the same complex shape but contained different ranges of disparity were measured. In the second experiment the minimum times required by experienced observers to perceive a given complex shape in stereograms that contained different ranges of disparity were determined. Perception times for the naive observers were all very fast (<3 s) and showed no evidence of perceptual learning. There was no effect of disparity range on perception times in either experiment. It was found that very large-disparity (80 min arc) complex stereograms could be perceived quickly, even by naive observers. It is concluded that the long initial latencies previously reported are not due to surface complexity nor to the range of disparities present. Other factors. such as dot size, dot density, and the correlation of the stereo images, appear to be important determinants of efficient stereoscopic performance when viewing complex random-dot stereograms. PMID- 8710439 TI - Transparent surfaces and illuminated holes. AB - A single stimulus determined the alternative perceptions of an illusory transparent grey disk or of an internally illuminated circular hole. A square on a far background was visible through this disk or hole. Subjects rated the grey colour of the transparent disk or the phenomenal illumination inside the hole. The luminance difference relative to the transparent disk and the square and that relative to this disk and its background determined the probability of perceiving the transparent disk or the hole. Rated colour and illumination substantially depended only on this second difference. These results have implications for models of phenomenal transparency and illumination based on the idea that proximal contours activate neural representations of phenomenal attributes. PMID- 8710440 TI - Texture discrimination at the cyclopean retina. AB - One of the open questions within the study of texture discrimination is whether the underlying neural mechanisms are located within the two monocular pathways, or whether they are located at more central areas that process binocular information. This question is considered here in psychophysical experiments of texture discrimination involving stereograms. The results show that texture discrimination for differences in form and size occur after binocular fusion. Moreover, the suitability of random-dot stereograms versus line-figure stereograms for this research has been studied. It was found that discrimination for differences in form was better with line-figure stereograms whereas discrimination for differences in size was better with random-dot stereograms. PMID- 8710441 TI - Perception of simple and complex visual stimuli: decision strategies and hemispheric differences in same-different judgments. AB - The accuracy with which observers could judge whether two visual stimuli were the same or different was measured with the rating method of detection theory. For judgments of whether two pictures referred to natural or manufactured things, the shape of the obtained receiver operating characteristic (ROC) was consistent with the observers adopting an optimal decision strategy. A similar result was found for judgments of complex but meaningless visual patterns. For judgments of whether two colours that differed along a simple sensory dimension were the same or different, however, the resulting ROC was consistent with the observers adopting a suboptimal differencing strategy. The accuracy of the judgments did not depend on the visual field to which the stimuli were presented. PMID- 8710442 TI - Vision comes to mind. AB - An overview is presented in which visual awareness is regarded as the epitome of the general mind-body problem. An experimental solution of the problem is considered first, followed by a philosophical outlook. It is argued that the scientific approach may eventually discover the neural correlate of visual awareness, but visual perceptions, even simple qualia like 'brightness' or 'color', occur in the mind of a conscious observer and are not reducible to observable activity of a specific set of neurons in the brain. PMID- 8710443 TI - On the filling in of the visual blind spot: some rules of thumb. AB - In monocular viewing there is a region in the peripheral visual field that is blind owing to the absence of photoreceptors at the site where the optic nerve exits the eye. This region, like certain other blind spots, nonetheless appears filled in. Several novel demonstrations of filling in at the blind spot have recently been reported. Here the implications of many of these effects are critically reevaluated. Specifically, it is argued that many blind-spot phenomena taken to support early filling in (eg pop out and alteration in apparent motion) are actually consistent with the thesis that the visual blind spot is treated by early perceptual processing as a region of reduced or absent information. In support of this, it is shown that many perceptual effects observed in blindspot completion are similar in detail to the amodally perceived completion of partly occluded objects viewed somewhat peripherally. The goals were to point out striking similarities between blind-spot completion and the amodal completion of occluded parts of surfaces, and to provide a common theoretical framework for understanding these phenomena in the context of surface segregation and perceptual interpolation. PMID- 8710444 TI - Filling in gaps in logic: reply to Durgin et al. PMID- 8710445 TI - Involuntary attentional shifts due to orientation differences. AB - We tested the ability of orientation differences to cause involuntary shifts of visual attention and found that these attentional shifts can occur in response to an orientation "pop-out" display. Texture-like cue stimuli consisting of discrete oriented bars, with either uniform orientation or containing a noninformative orthogonally oriented bar, were presented for a variable duration. Subsequent to or partially coincident with the cue stimulus was the target display of a localization or two-interval forced-choice task, followed by a mask display. Naive subjects consistently showed greater accuracy in trials with the target at the location of the orthogonal orientation compared with trials with uniformly oriented bars, with only 100 msec between the cue and mask onsets. Discriminating these orientations required a stimulus onset asynchrony (SOA) of 50-70 msec. The attentional facilitation is transient, in most cases absent with a cue-mask SOA of 250 msec [corrected]. These results suggest that the preattentive character of some texture discrimination tasks with SOAs of only 100 msec is vitiated by the involuntary attentional shifts that are caused by orientation differences. PMID- 8710446 TI - The visual perception of rigid motion from constant flow fields. AB - Four experiments investigated observers' judgments of rigidity for different types of optical motion. The depicted structural deformations were of two types: (1) those with nonparallel image trajectories that are detectable from the first order spatiotemporal relations between pairs of views; and (2) those with parallel image trajectories that can only be detected from higher order relations among three or more views. Patterns were composed of smooth flow fields in Experiments 1 and 3, and of wire frame figures in Experiments 2 and 4. For both types of display, the nonrigidity detectable from the first-order spatiotemporal structure of the motion sequence was much more salient than the deformation detectable only from the higher order spatiotemporal structure. These results indicate that observers' judgments of rigidity are based primarily on a two-view analysis, but that some useful information can be obtained under appropriate circumstances from higher order spatiotemporal relations among three or more views. PMID- 8710447 TI - Discrimination of vibrotactile frequencies in a delayed pair comparison task. AB - This study quantified human short-term-memory decay functions for delayed vibrotactile frequency discriminations. Subjects indicated which of two successive intervals contained the higher or lower frequency of a pair separated by delay periods of 0.5-30 sec. Performance decreased as a function of length of delay and was higher when delays were unfilled than when they were filled with a backwards-counting task. This interpolated task may have interfered with rehearsal of a coded representation of the remembered vibrotactile frequency. A change in decay rate after 5-sec delays suggests a switch from reliance on sensory memory to the coded frequency representation. Performance and decay rate depended on presentation order of higher or lower frequency within pairs. Reciprocal performance asymmetries seen in high-versus low-frequency ranges did not result from simple response bias. PMID- 8710449 TI - Effects of harmonics on relative pitch discrimination in a musical context. AB - The contribution of different harmonics to pitch salience in a musical context was examined by requiring subjects to discriminate a small (1/4 semitone) pitch change in one note of a melody that repeated in transposition. In Experiment 1, performance was superior when more harmonics were present (first five vs. fundamental alone) and when the second harmonic (of tones consisting of the first two harmonics) was in tune compared with when it was out of tune. In Experiment 2, the effects of harmonics 6 and 8, which stand in octave-equivalent simple ratios to the fundamental (2:3 and 1:2, respectively) were compared with harmonics 5 and 7, which stand in more complex ratios (4:5 and 4:7, respectively). When the harmonics fused into a single percept (tones consisting of harmonics 1, 2, and one of 5, 6, 7, or 8), performance was higher when harmonics 6 or 8 were present than when harmonics 5 or 7 were present. When the harmonics did not fuse into a single percept (tones consisting of the fundamental and one of 5, 6, 7, or 8), there was no effect of ratio simplicity. PMID- 8710448 TI - An investigation of the gate control theory of pain using the experimental pain stimulus of potassium iontophoresis. AB - This study investigated a prediction derived from gate control theory-that there would be a pulse of pain as a pain stimulus was being ramped off due to the rapidly transmitting, inhibitory large fiber activity falling away sooner at the spinal level than the excitatory activity of the slow-transmitting, small nociceptive afferents. A further prediction was that the more distant the peripheral stimulus was from the spine, the greater the pain pulse would be. Fourteen subjects had the pain stimulus of iontophoretically applied potassium ions (K+) applied to an upper and a lower site on the dominant arm. In a threshold detection task using the double random staircase method, subjects were asked to indicate whether they could detect a pulse of additional pain during this ramp-off phase. The average rate of stimulus ramp-off in order to detect a pain pulse was statistically greater for the upper-arm site (14.3 micrograms K+/sec) than for the lower-arm site (9.4 micrograms K+/sec). These results were consistent with gate control theory. Alternative explanations, including intrinsic differences in nociceptive responding for different dermatomes and anode break, were considered. It was concluded that the detection of a pain pulse during the ramping off of a peripheral pain stimulus potentially provides a quantitative measure of the spinal modulation of pain. PMID- 8710451 TI - The missing fundamental in vowel height perception. AB - Traunmuller (1981) suggested that the tonotopic distance between the first formant (F1) and the fundamental frequency (F0) is a major determinant of perceived vowel height. In the present study, subjects identified a vowel-height continuum ranging in formant pattern from /I/to/epsilon/, at five F0 values. Increasing F0 led to an increased probability of /I/responses (i.e., the phoneme boundary shifted toward the /epsilon/ end of the continuum). Various conditions of filtering out the lower harmonics of the stimuli caused only marginal shifts of the phoneme boundary. The experiments provide evidence against interpretations of Traunmuller's (1981) results that claim that vowel height is determined by the distance between F1 and the lowest harmonic that is present in the basilar membrane excitation pattern. PMID- 8710450 TI - Determinants of cumulative successive contrast in saltiness intensity judgments. AB - When all stimuli elicit the same taste quality, solutions preceded by a high concentration level are judged to be significantly less intense than solutions preceded by a low concentration level. After repetitious stimulation with a different tasting stimulus, the intensity of the present stimulus is over estimated. This phenomenon is called "successive contrast." In the present study, the cumulative effects of three identical stimuli on the saltiness ratings for a test stimulus are investigated. The preceding stimuli are manipulated with regard to taste quality, saltiness intensity, total taste intensity, and complexity. Whether the size of the cumulative contrast effect is associated with the degree of dissimilarity between preceding stimuli and test stimulus, or with the saltiness or total taste intensity of the preceding stimuli, is investigated. The size of the contrast effect depends on the type of preceding stimulus, its intensity, and the type of test stimulus. No association was found with judgments of the degree of dissimilarity between the preceding stimuli and the test stimulus. For nonsalty preceding stimuli, the contrast effects are independent of concentration level. When the preceding stimuli taste at least partly salty, the total intensity appears to determine the size of the contrast for an unmixed salty test stimulus. PMID- 8710452 TI - Mindless reading revisited: eye movements during reading and scanning are different. AB - In an extension of a study by Vitu, O'Regan, Inhoff, and Topolski (1995), we compared global and local characteristics of eye movements during (1) reading, (2) the scanning of transformed text (in which each letter was replaced with a z), and (3) visual search. Additionally, we examined eye behavior with respect to specific target words of high or low frequency. Globally, the reading condition led to shorter fixations, longer saccades, and less frequent skipping of target strings than did scanning transformed text. Locally, the manipulation of word frequency affected fixation durations on the target word during reading, but not during visual search or z-string scanning. There were also more refixations on target words in reading than in scanning. Contrary to Vitu et al.'s (1995) findings, our results show that eye movements are not guided by a global strategy and local tactics, but by immediate processing demands. PMID- 8710453 TI - Perceptual integrality of major chord components. AB - In the present study, an accuracy, rather than a reaction time, version of the Garner paradigm was used to evaluate the integrality or separability of major chord components. Tuned (prototype, or P) and mistuned (nonprototype, or NP) sets of root position C-major triads were constructed by holding the C constant in all stimuli and varying the E and G frequencies in 2- and 4-Hz steps. The P stimuli represent small systematic mistunings in the E and G notes relative to an equal tempered C-major chord. The NP stimuli represent an equivalent range of frequency variation, but relative to a significantly out-of-tune C-major triad. In different experimental sessions, a same-different (AX) task was used to separately evaluate discrimination performance for the E and G frequencies as a function of whether the nontarget frequency (G or E) was fixed or varied in either a correlated or an orthogonal fashion (with the C frequency always held constant). Compared with a fixed baseline condition where only the target frequency changed, both chord components exhibited a significant redundancy gain in the correlated conditions and, to varying degrees, significant interference effects in the orthogonal condition, indicating that the chord components function largely in an integral fashion. Relative to the discrimination of G, discrimination of the E frequency was less influenced by variation in the nontarget (G) frequency, showing that attention, to some degree, could be selectively allocated to the E chord component. In addition, the results were consistent with previous findings that the functional prototype for the major chord category seems to act as a perceptual anchor, rather than as a magnet, and appears to be located in the physiologically defined area of just temperament, as opposed to the more experientially defined area of equal temperament. PMID- 8710454 TI - Perception of local orientation from shaded images. AB - The perception of local orientation from shaded images was examined. In Experiment 1, subjects viewed a boundaryless Gaussian hill and judged local orientation using both a gauge figure and a pointing method. One subject reported an internally consistent surface which was incompatible with the judged light source direction and model used to generate the image. The remaining subjects reported a surface similar to the generating one, and analysis of their results indicated a contour of zero difference between response and generating slants. This contour of zero slant difference was explored in three subsequent experiments using the pointing technique. These experiments investigated possible influences of luminance artifact (Experiment 2), perception of global orientation (Experiment 3), and self-occluding contours (Experiment 4). All three of these experiments yielded results similar to those of Experiment 1, with distinct contours of zero slant difference. This contour was explored for relationships with the simulated slant of the generating surface and the differential structure of image intensity. This analysis indicated that the contour of zero slant difference was approximately a line of constant slant which shared large regions of adjacency to the zero crossings of the second directional derivative of image intensity. PMID- 8710455 TI - The nature and duration of adaptation following long-term odor exposure. AB - Any individual living or working in an odorous environment can experience changes in odor perception, some of which are long lasting. Often, these individuals report a significant reduction in the perception of an odor following long-term exposure to that odor (adaptation). Yet, most experimental analyses of olfactory adaptation use brief odorant exposures which may not typify real-world experiences. Using a procedure combining long-term odor exposure in a naturalistic setting with psychophysical tests in the laboratory, we present evidence to show that reduced odor intensity following long-term exposure is accompanied by odorant-specific shifts in threshold. Subjects were exposed continuously to one of two odorants while in their home for a period of 2 weeks. Exposure produced an odorant-specific reduction in sensitivity and perceived intensity compared with preexposure baselines: Detection thresholds for the adapting odorant were elevated following exposure and perceived intensity ratings for weak concentrations were reduced. For most individuals, reduced sensitivity to the test odorant was still evident up to 2 weeks following the last exposure. The persistence of the change, as evidenced by the duration of recovery from adaptation, distinguishes this phenomenon from the adaptation seen following shorter exposures and highlights the need for the study of exposure durations that are more similar to real-world exposures. PMID- 8710457 TI - [Shunt patency after creation of a transjugular intrahepatic portosystemic shunt: evaluation with Doppler ultrasound]. AB - TIPS (transjugular intrahepatic portosystemic shunt) is a new interventional radiologic procedure that is useful in the treatment of variceal bleeding and ascites due to portal hypertension. It is most important that the shunt remains patent for a long time after placement. The potential role of ultrasound in evaluating the patency of the shunt after TIPS has been studied. Duplex and color doppler sonography were performed in 20 patients with portal hypertension after TIPS. Imaging was done in 38 case. The findings were compared with those of follow-up angiography. Shunt patency was evaluated with Doppler US in 28 case. In 27 of them, the equivalent patent shunt was confirmed by angiography; the exception was one case of occlusion in follow-up angiography. Doppler signals in the shunt could not be detected in 10 cases. Follow-up angiography showed occlusion in 7 cases and patency in the other 3. Doppler US sensitivity was 90%, and its specificity was 88%. These results indicate that Doppler US is one of the most useful methods for evaluating shunt patency after TIPS. PMID- 8710456 TI - Intensity resolution and subjective magnitude in psychophysical scaling. AB - Several successful theories of psychophysical judgment imply that exponents of power functions in scaling tasks should covary with measures of intensity resolution such as d' in the same tasks, whereas the prevailing metatheory of ideal psychophysical scaling asserts the independence of the two. In a direct test of this relationship, three prominent psychophysical scaling paradigms were studied: category judgment without an identification function, absolute magnitude estimation, and cross-modality matching with light intensity as the response continuum. Separate groups of subjects for each scaling paradigm made repeated judgments of the loudnesses of the pure tones that constituted each of two stimulus ensembles. The narrow- and wide-range ensembles shared six identical stimulus intensities in the middle of each set. Intensity resolution, as measured by d'-like distances, of these physically identical stimuli was significantly worse for the wide-range set for all three methods. Exponents of power functions fitted to geometric mean responses, and in magnitude estimation and cross modality matching the geometric mean responses themselves, were also significantly smaller in the wide-range condition. The variation of power function exponents, and of psychophysical scale values, for stimulus intensities that were identical in the two stimulus sets with the intensities of other members of the ensembles is inconsistent with the metatheory on which modern psychophysical scaling practice is based, although it is consistent with other useful approaches to measurement of psychological magnitudes. PMID- 8710458 TI - [MR imaging in patients with bone marrow metastases of neuroblastoma]. AB - Sixty patients with neuroblastoma or ganglioneuroblastoma underwent MRI 87 procedures to detect and follow up bone marrow metastases. The detection of bone marrow metastases by MRI was evaluated with aspiration cytology. Relationships were investigated between bone metastases and bone marrow metastases that were morphologically classified as nodular and diffuse by MRI findings. The usefulness of MRI was also studied in evaluating the effect of chemotherapy. MRI showed bone marrow metastases in all of 17 patients, but aspiration cytology proved metastases in only 7 of 17. Bone metastases were proven by bone scintigraphy or bone X-ray in 5 of 15 patients, and only diffuse bone marrow metastases were accompanied with bone metastases. After or during chemotherapy, 10 patients were examined by MRI more than two times. Bone marrow metastases disappeared in 9, decreased in size in 3, did not change in one and recurred at another site in one. It was observed from MRI findings that in the early stage, bone marrow metastases are nodular, but along with progress of the disease, they become diffuse and invade the bone cortex. If chemotherapy was effective, bone marrow metastases were decreased in size or disappeared completely. PMID- 8710459 TI - [Use of oblique sagittal T2-weighted images in diagnosing rotator cuff tears]. AB - The correlation between oblique sagittal T2-weighted images and oblique coronal T2-weighted images was determined in 86 patients who had symptoms and signs suggesting rotator cuff tears. Abnormal high signal lesions suggesting full and partial thickness tear of the supraspinatus tendon were observed on the oblique coronal images in 20 and 27 of 86 patients, respectively. The oblique sagittal images showed the lesions just the same as the oblique coronal images. In the other 39, abnormal high signal lesions were not observed on oblique coronal images or on oblique sagittal images. The oblique sagittal images were proved not useful to detect tears of the supraspinatus tendon. However, the oblique sagittal T2-weighted images showed posterior extension of high signal lesions and clarified abnormal high signal lesions near the long head of the biceps tendon in two patients. These results suggested that oblique coronal images should always be taken at the first step to identify supraspinatus tendon tear, and oblique sagittal images can be taken as an adjust to the former images. PMID- 8710461 TI - Can MRI distinguish between acute partial and complete anterior cruciate ligament tear? PMID- 8710460 TI - [Relation between anterior displacement of the temporomandibular joint disc and size of the condyle]. AB - To elucidate the cause of anterior displacement of the disc, we evaluated the relation between anterior displacement and the size of the condyle from MR images in 301 patients (602 TMJs) with clinical diagnosis of internal derangement. We found that the size of condyles with anterior displacement of the disc was significantly smaller than that of condyles without anterior displacement. This result suggests that small condyles may give rise to anterior displacement of the disc. PMID- 8710462 TI - [Brain CT and MRI findings in fat embolism syndrome]. AB - To elucidate brain CT and MRI findings in fat embolism syndrome (FES), we retrospectively analyzed images from 5 patients with FES during the acute and subacute stages. Brain CT examinations demonstrated brain edema in 2 patients and transient spotty low density lesions in 2 patients. Three patients showed no abnormalities. Brain MRI, however, showed brain abnormalities in all patients during the acute stages. These were revealed as spotty high signal intensity lesions on T2WI, and some showed low intensity on T1WI. These spotty lesions were considered to reflect edematous fluid occurring as a result of the unique pathophysiological condition of FES. While the spotty high signal intensity lesions on T2WI were distributed in the cerebrum, cerebellum, brain stem, thalamus, basal ganglia, internal capsule and corpus callosum, cerebral and cerebellar spotty lesions were characteristically located along the boundary zones of the major vascular territories. This characteristic location might be induced by a hypoxic brain condition in FES because the numerous fat globules present in this condition can block entire brain capillaries. This characteristic signal location on T2WI is a useful indicator for differentiating FES from the primary intra-axial brain injury in patients with multifocal trauma. PMID- 8710463 TI - [Clinical evaluation of power Doppler sonography for the diagnosis of hepatocellular carcinoma]. AB - We compared power Doppler sonography (PDS) with conventional color Doppler sonography (CDS) in the visualization of intratumoral blood flow of hepatocellular carcinoma (HCC) and evaluated the usefulness of PDS for assessing of the effects of TAE with Lipiodol and anticancer agents (Lp-TAE) and for the diagnosis of recurrence of HCC after Lp-TAE. The ability of PDS and CDS to visualize intratumoral blood flow was compared in 36 patients with HCC, and the usefulness of PDS for the diagnosis of HCC was evaluated. Also, in 17 patients with HCC who underwent Lp-TAE, the usefulness of PDS for evaluating the therapeutic effects of Lp-TAE and the diagnosis of recurrence after Lp-TAE was examined. The ability of PDS to visualize intratumoral blood flow and delineate tortuous and meandering intratumoral vasculature was superior to that of CDS. The effects of Lp-TAE could be evaluated and recurrence detected accurately and easily by PDS on the basis of residual intratumoral blood flow and the appearance of new blood flow. PDS is expected to achieve widespread acceptance as a useful imaging modality for the diagnosis of HCC, assessment of therapeutic effects of TAE, and detection of recurrence of HCC after TAE. PMID- 8710465 TI - [Usefulness of endoscopic ultrasonography in deciding indication for high dose rate intracavitary brachytherapy for esophageal carcinoma]. AB - We retrospectively analyzed 20 patients with stage I-IV esophageal carcinoma treated by limited external radiotherapy (EXR) combined with high dose rate intracavitary brachytherapy (ICBT) using 60Co as boost therapy. In 10 of 20 patients treated by combined therapy, we used endoscopic ultrasonography (EUS) before ICBT and measured the thickness of the residual tumor following external beam therapy. Of the patients treated by combined therapy, the local control rate of esophageal carcinoma with a thickness under 10 mm before ICBT was significantly better than that with a thickness over 10 mm (confidence limit 95%). Therefore, when administering high dose rate intracavitary brachytherapy after EXR, we should measure the thickness of esophageal carcinoma with millimeter unit precision. One of the most precise methods currently available for measuring thickness is EUS. Our results indicate that esophageal carcinoma under 10 mm in thickness after EXR is the most favorable indication for ICBT. PMID- 8710464 TI - [Age-dependent changes in metabolites of the normal brain in childhood : observation by proton MR spectroscopy]. AB - We investigated aging-dependent changes in proton magnetic resonance spectroscopy (1H-MRS) of the normal brain in childhood, and observed differences in the four portions of the brain. Measurement by 1H-MRS was carried out on the frontal lobe, parietotemporal lobe, temporal lobe and cerebellum. The NAA/Cho ratio increased rapidly in the period from 0 to 2 years of age in all portions except for the cerebellum, and gradually increased after three years of age. The number of measurements of the cerebellum was not sufficient to reach a conclusion, but no clear aging-related change was found. The Cho/Cr ratio decreased according to the neural development in all portions except the cerebellum. Because the T2 relaxation time of water after four years of age was almost the same as that of young adults, we used the relaxation times specified in the literature to quantify the metabolites observed by 1H-MRS. The subjects used for quantification were aged from 4 to 12 years. The concentration of NAA in the temporal lobe was the lowest of the four portions, and that of Cho and Cr in the cerebellum was the highest in four portions. These results could not be obtained by signal ratios alone, and we considered that the quantification of metabolites is necessary for a better understanding of 1H-MRS. This study showed that the results of 1H-MRS vary depending on age and the portion in the brain. Our results may serve as a normal basis for the detection of pathological changes by 1H-MRS. PMID- 8710466 TI - [Analysis of locally controlled esophageal carcinomas treated with radiotherapy]. AB - Of 227 esophageal carcinomas treated with a radiation dose of 60 Gy or more, 100 patients had no tumor or ulceration (with or without stenosis) of the esophagus after irradiation. We analyzed local control factors of these 100 patients to determine the need for further treatment. The cumulative local control rate at five years was 40% in all cases, 37% in 21 cases without any stenosis of the esophagus and 40% in 79 cases with stenosis. The presence of stenosis of the esophagus after irradiation was not a critical factor in predicting final local control. Local recurrence of tumors with findings of Borrmann III or Borrmann IV by the pretreatment esophageal barium study, tumors controlled after a total dose of more than 80 Gy, tumors without low dose rate telecobalt therapy (LDRT; 1 Gy/hour, 5 to 7Gy/day, a total dose of 12 to 15 Gy) as boost therapy, and apparently controlled tumors with a stenotic ratio of 60% or more or with 5 cm or more length of stenosis of the esophagus after irradiation was significantly higher than that of the others (p < 0.05). Multivariate analysis revealed that findings of pretreatment barium study, total dose, with or without LDRT, and length of stenosis of the esophagus after irradiation were significantly important factors in local control. Members of the high risk group of apparently controlled tumors should undertake surgical treatment or further intensive chemotherapy. PMID- 8710467 TI - [Palliation of brain metastases: factors associated with survival]. AB - One hundred ten patients with brain metastases (76 lung cancer, 11 breast cancer, 7 colorectal cancer and others) were treated with radiotherapy, and the results were retrospectively evaluated. Neurologic symptoms were improved in 66% of patients. The relationship between improvement of neurologic symptoms and several factors, including irradiation dose, primary lesion, histology, term between first treatment and recurrence, clinical symptoms and CT findings, was analyzed. There was no significant correlation between symptom relief and these factors except for the number of metastatic lesions. Four of 110 patients could survive more than 2 years after radiotherapy: one with lung cancer, one with breast cancer, one with rectum cancer and one with uterine cancer. The factors that were analyzed for symptom relief were also studied to determine their effect on the survival of lung cancer patients with brain metastases. The patients who had higher dose irradiation (50 Gy) of improvement of neurological symptoms after radiotherapy survived significantly longer than those who had 30 Gy of irradiation or no symptom relief. Multivariate analysis of these factors showed that improvement of symptoms and irradiation dose were similarly important prognostic factors, whereas the others were not correlated with survival. PMID- 8710468 TI - [Evaluation of image quality of medical facsimile as a radiological image recording system]. AB - We examined the usefulness of a new medical facsimile (MFAX) system in recording and transmitting various kinds of medical images, including X-ray images and colored histopathologic images. The system consists of an image scanner, a magnetic disk for image storage, a transmission circuit and a thermal image printer. Transmission time for a FCR (Fuji computed radiography) image by super fine mode was 6 minutes. We used ROC (receiver operating characteristics) curves to evaluate the ability of eight radiologists to detect the small simulated nodules placed on an anthropomorphic chest phantom and shown on MFAX images. The radiologists observed both the FCR films and MFAX copies and determined the presence or absence of simulated nodules using five confidence levels. The results obtained for FCR films and MFAX images showed no statistically significant difference. PMID- 8710469 TI - [Simplified radiographic slide production utilizing a laser imager and personal computer]. AB - A simple, rapid, inexpensive method for producing radiographic slides is described. By connecting the divided monitor output of a personal computer to a laser imager in the Radiology Department, radiographics and text displayed on the personal computer could be printed out from the laser imager. By selecting a print format of slide size, 12 slide-sized images could be printed on film. In addition, by selecting other print formats, various sized hard-copy images could be obtained. These print-outs could only be produced in black and white, but were convenient for radiology conferences and teaching files. PMID- 8710470 TI - A study of student retention in associate degree nursing programs as perceived by their directors. PMID- 8710471 TI - Gateway to the caregiver: collaborating to meet the needs of education, practice, and community. PMID- 8710472 TI - The Healing Web: curriculum changes to support differentiated practice roles in the community. PMID- 8710473 TI - Community-based maternal child nursing. PMID- 8710474 TI - Senior students and senior citizens: reciprocal benefits in learning and caring. PMID- 8710475 TI - Vision for nursing education reform. PMID- 8710476 TI - The web of inclusion: faculty helping faculty to weave the fabrics of our future. PMID- 8710477 TI - Second Annual Mildred Montag Excellence in Leadership Award: the founding directors of the first associate degree nursing programs. PMID- 8710478 TI - Positioning health care programs for the future. PMID- 8710479 TI - Designing a program evaluation plan that complements the NLN self-study criteria: outcome collaboration. PMID- 8710480 TI - Mentoring: making a difference in nursing education. PMID- 8710481 TI - Leadership and collaboration: sharing of power. PMID- 8710482 TI - Integrity, access, and accountability: health care reform's new ethical dilemmas. PMID- 8710483 TI - Creating an environment for faculty change: how one faculty member joined the curriculum revolution and lived to tell about it. PMID- 8710484 TI - Caring groups: an experiential teaching/learning strategy. PMID- 8710485 TI - Bridging the gap in generational understanding: shared learning between the well elderly client and nursing students. PMID- 8710486 TI - Compilation and classification of higher plant mitochondrial tRNA genes. AB - This compilation reports the tRNA genes detected on higher plant mitochondrial genomes subdivided into the widely accepted categories of 'genuine' and 'chloroplast-like' genes. Moreover, it includes a list of pseudo or truncated genes divided in the same way. PMID- 8710487 TI - Common DNA structural features exhibited by eukaryotic ribosomal gene promoters. AB - Nucleotide sequences of DNA regions containing eukaryotic ribosomal promoters were analysed using strategies designed to reveal sequence-directed structural features. DNA curvature, duplex stability and pattern of twist angle variation were studied by computer modelling. Although ribosomal promoters are known to lack sequence homology (unless very closely related species are considered), investigation of these structural characteristics uncovered striking homologies in all the taxonomic groups examined so far. This wide conservation of DNA structures, while DNA sequence is not conserved, suggests that the determined structures are fundamental for ribosomal promoter function. Moreover, this result agrees well with the recent observations showing that RNA polymerase I transcription factors have not evolved as intensively as previously suspected. PMID- 8710488 TI - Another heritage from the RNA world: self-excision of intron sequence from nuclear pre-tRNAs. AB - The intervening sequences of nuclear tRNA precursors are known to be excised by tRNA splicing endonuclease. We show here that a T7 transcript corresponding to a pre-tRNA(Tyr) from Arabidopsis thaliana has a highly specific activity for autolytic intron excision. Self-cleavage occurs precisely at the authentic 3' splice site and at the phosphodiester bond one nucleotide downstream of the authentic 5'-splice site. The reaction results in fragments with 2',3'-cyclic phosphate and 5'-OH termini. It is resistant to proteinase K and/or SDS treatment and is not inhibited by added tRNA. The self-cleavage depends on Mg2+ and is stimulated by spermine and Triton X-100. A set of sequence variants at the cleavage sites has been analysed for autolytic intron excision and, in parallel, for enzymatic in vitro splicing in wheat germ S23 extract. Single-stranded loops are a prerequisite for both reactions. Self-cleavage not only occurs at pyrimidine-A but also at U-U bonds. Since intron self-excision is only about five times slower than the enzymatic intron excision in a wheat germ S23 extract, we propose that the splicing endonuclease may function by improving the preciseness and efficiency of an inherent pre-tRNA self-cleavage activity. PMID- 8710489 TI - Characterisation of antibody-binding RNAs selected from structurally constrained libraries. AB - Constrained RNA libraries of limited sequence complexity were constructed and used to select RNA molecules binding to the antigen binding site of an anti ferritin antibody. The sequences required as primer-binding sites for the selection cycle were designed to form a predictable secondary structure, which greatly facilitated the characterisation of the secondary structures of the selected RNAs. RNA-antibody interactions were studied by real-time interaction analysis to study the dynamic aspects of binding and by circular dichroism spectroscopy to search for conformational changes upon binding. The selected RNAs were analysed with a binding site sequestering assay and were shown to compete with ferritin for binding to the antigen-binding site. The experiments described here indicate that the introduction of strong structural constraints does not have to interfere with the ability to select tightly and specifically binding RNA molecules. PMID- 8710490 TI - Interaction of mRNA with the Escherichia coli ribosome: accessibility of phosphorothioate-containing mRNA bound to ribosomes for iodine cleavage. AB - The contacts of phosphate groups in mRNAs with ribosomes were studied. Two mRNAs were used: one mRNA contained in the middle two defined codons to construct the pre- and the post-translocational states, the other was a sequence around the initiation site of the natural cro-mRNA. Phosphorothioate nucleotides were randomly incorporated at a few A, G, U or C positions during in vitro transcription. Iodine can cleave the thioated positions if they are not shielded by ribosomal components. Only a few minor differences in iodine cleavage of ribosome bound and non-bound mRNA were observed: the nucleotide two positions upstream of the decoding codons (i.e. those codons involved in codon-anticodon interactions) showed a reduced accessibility for iodine and the nucleotide immediately following the decoding codons an enhanced accessibility in both elongating states. In initiating ribosomes where the mRNA contained a strong Shine-Dalgarno sequence, at least five phosphates were additionally slightly protected covering the Shine-Dalgarno sequence and nucleotides downstream including the initiator AUG in the P site (Al, G3, G-2, G-5 and A-7). The low contact levels of the phosphates in the mRNA with the elongating ribosome strikingly contrast with the pronounced contact patterns previously described for tRNAs. The data obtained in this study, as well as results of previous studies, suggest that mRNA regions downstream and upstream of decoding codons form only weak contacts with ribosomal components and that the mRNA thus is mainly fixed by codon-anticodon interaction on the elongating ribosome. PMID- 8710491 TI - Inhibition of NF-kappaB DNA binding by nitric oxide. AB - It has been suggested that the NF-kappaB transcription factor family may mediate expression of the gene encoding the cytokine-inducible form of nitric oxide synthase (iNOS). To establish if nitric oxide (NO) could in turn affect activity of NF-kappaB, the ability of NO-donor compounds to influence NF-kappaB DNA binding activity in vitro was investigated. NO-donor compounds sodium nitroprusside (SNP) and S-nitroso-N-acetylpenicillamine (SNAP) both inhibited the DNA binding activity of recombinant NF-kappaB p50 and p65 homodimers and of p50 p65 heterodimers. Inhibition of NF-kappaB p50 DNA binding by NO-donor compounds involved modification of the conserved redox-sensitive C62 residue, as a C62S p50 mutant was significantly more resistant to SNP-mediated inactivation. Non reducing SDS-polyacrylamide gel electrophoresis demonstrated that SNP could inhibit p50 DNA binding by mechanisms other than the formation of intersubunit disulphide bonds involving p50 residue C62. Electrospray ionization mass spectrometry of a synthetic NF-kappaB p5O peptide containing the C62 residue suggested that NO gas can modify C62 by S-nitrosylation. This study indicates that NO-donors can directly inhibit the DNA binding activity of NF-kappaB family proteins, suggesting that cellular NO provides another control mechanism for modulating the expression of NF-kappaB-responsive genes. PMID- 8710492 TI - Location of the C-terminal domain of the RNA polymerase alpha subunit in different open complexes at the Escherichia coli galactose operon regulatory region. AB - Hydroxyl radical footprinting has been used to study different open complexes between Escherichia coli RNA polymerase and the galactose operon regulatory region, which contains two overlapping promoters, P1 and P2. Complexes at P1 were studied by exploiting a P2- mutant and complexes at P2 were studied with a P1 mutant. We have identified the precise location of alpha binding in both binary RNA polymerase-galP1 and RNA polymerase-P2 complexes from the effects of deletion of the C-terminal domain of the RNA polymerase alpha subunit: alpha binds to different sites at the upstream end of each complex. Transcription initiation at galP1 can be activated by the cyclic AMP receptor protein (CRP). Addition of CRP to the RNA polymerase-galP1 complex displaces the C-terminal domain of alpha, which then binds to a different site upstream of CRP in the ternary CRP-RNA polymerase-galP1 complex. Thus, the C-terminal domain of alpha can occupy three different sites at the gal operon regulatory region. We have also examined the effect of disrupting the Activating Region of CRP on interactions between CRP and the C-terminal domain of alpha in ternary CRP-RNA polymerase-galP1 complexes. Footprinting experiments show that these substitutions interfere with the contact between CRP and alpha but do not affect the position of alpha binding to its site upstream of bound CRP. PMID- 8710493 TI - Transactivation by the thyroid hormone receptor is dependent on the spacer sequence in hormone response elements containing directly repeated half-sites. AB - The thyroid hormone receptor (TR) regulates the transcription of its target genes by interacting with specific hormone response elements consisting usually of directly repeated half-sites with the consensus sequence AGGTCA. To investigate the role of the spacer sequences separating the half-sites, heterodimers formed by TRalpha and the retinoid-X receptor (RXR) were used in a PCR based selection and amplification assay. The TRalpha/RXR heterodimer selected for elements with directly repeated half-sites having a spacer of 4 nucleotides (DR4). Preferences for nucleotides in the TR binding half-site motif as well as for the 4 nucleotides separating the two half-sites were found. DNA binding and transfection studies using DR4 elements with different spacer sequences showed the importance of these nucleotides for the activity of the response element: some spacer sequences allowed little or no transactivation from the element, whereas other sequences supported strong transactivation. A pyrimidine nucleotide in position three of the spacer enhanced TRalpha binding and transactivation. Additional experiments showed that heterodimers between RXR and other putative receptors exhibited a similar but distinct specificity for the spacer sequence. Our results thus suggest that the four nucleotides separating the two half-sites in hormone response elements have a major role in determining induction of hormone responsive genes. PMID- 8710494 TI - Modelling extreme stretching of DNA. AB - Molecular modelling with Jumna is used to study extreme stretching of the DNA double helix. The results, which correlate well with recent nanomanipulation experiments, show how the double helix can be extended to twice its normal length before its base pairs break. Depending on the way the duplex is stretched two types of conformation can occur, either an unwound flat ribbon or a narrow fibre with negatively inclined base pairs. The energetics of both types of deformation are similar and existing structures show that at least the flat ribbon form can exist locally under biological conditions. PMID- 8710495 TI - Cloning and expression of the BalI restriction-modification system. AB - BalI, a type II restriction-modification (R-M) system from the bacterium, Brevibacterium albidum, recognizes the DNA sequence 5'-TGGCCA-3'. We cloned the genes encoding the BalI restriction endonuclease and methyltransferase and expressed them in Escherichia coli. The two genes were aligned tail-to-tail and their termination codons overlapped. BalI restriction endonuclease and methyltransferase comprise 260 and 280 amino acids, respectively, and have molecular weights of 29 043 and 31 999 Da. The amino acid sequence of BalI methyltransferase is similar to that of other m6A MTases, although it has been categorized as a m5C methyltransferase. A high expression system for the BalI restriction endonuclease was constructed in E. coli for the production of large quantities of enzyme. PMID- 8710496 TI - Replication of yeast DNA and novel chromosome formation in mouse cells. AB - To determine whether yeast DNA can replicate or segregate in mammalian cells, we have transferred genomic DNA from the yeast Saccharomyces cerevisiae into mouse cells. Most of the lines contained stably integrated yeast DNA. However, in two of the lines, the yeast DNA was maintained as numerous small extrachromosomal elements which were still present after 26 cell divisions in selection but which were lost rapidly out of selection. This indicates that, although yeast DNA can replicate in mouse cells, the yeast centromere does not function to give segregation. In one cell line we observed a large novel chromosome consisting almost entirely of yeast DNA. This chromosome segregates well and contains mouse centromeric minor satellite DNA and variable amounts of major satellite DNA which probably comprise the functional centromere. The yeast DNA in the novel chromosome has a compacted chromatin structure which may be responsible for the efficient formation of anaphase bridges. Furthermore, yeast DNA integrated into mouse chromosomes forms constrictions at the point of integration. These features have previously been presumed to be hallmarks of centromeric function in transfection assays aimed at identifying putative centromeric DNA. Hence our results suggest caution be exercised in the interpretation of such assays. PMID- 8710497 TI - Characterization of proteolytic fragments of bacteriophage T7 DNA ligase. AB - Treatment of T7 DNA ligase with a range of proteases generates two major fragments which are resistant to further digestion. These fragments, of molecular weight 16 and 26 kDa, are derived from the N- and C-termini of the protein, respectively. The presence of ATP or a non-hydrolysable analogue, ADPNP, during limited proteolysis greatly reduces the level of digestion. The N-terminal 16 kDa region of the intact T7 ligase is labelled selectively in the presence of [alpha 32P]ATP, confirming that it contains the active site lysine residue. In common with the intact enzyme, the C-terminal portion of the protein retains the ability to band shift DNA fragments of various lengths, implicating it in DNA binding. It can also inhibit ligation by the intact protein, apparently by competing for target sites on DNA. We conclude that the N-terminal region, which contains the putative active site lysine, plays a role in the transfer of AMP from the enzyme adenylate complex to the 5'phosphate at the nick site, while the C-terminal 26 kDa fragment appears to position the enzyme at the target site on DNA. PMID- 8710498 TI - Pbx-1 Hox heterodimers bind DNA on inseparable half-sites that permit intrinsic DNA binding specificity of the Hox partner at nucleotides 3' to a TAAT motif. AB - Heterodimers between the Pbx/Exd and Hox/HOM-C classes of homeodomain proteins bind regulatory elements in tissue-specific and developmentally regulated genes. In this work, we characterize the half-site bound by both Pbx1 and Hox proteins on a prototypic element (TGATTAAT) and determine how the orientation of the Hox protein contributes to the DNA binding specificity of Pbx-Hox heterodimers. We demonstrate that the Hox protein binds the 3' TAAT sequence as its recognition core and exhibits sequence-specific binding at positions 3' to the TAAT core. Unfavored sequences at this position, such as two cytosines, abrogate binding to the element. The upstream Pbx1 core sequence, TGAT, must immediately juxtapose the Hox core. This geometry maintains the preference of Hox/HOM-C proteins for a T base at position -1, as T represents the fourth position of the Pbx1 core, and suggests that this T base is bound by both Pbx1 and Hox proteins, Pbx1 binding in the major grove and the Hox protein binding in the minor grove. Pbx1 also exhibits base selectivity 5' to its TGAT recognition sequence. PMID- 8710499 TI - repE--the Dictyostelium homolog of the human xeroderma pigmentosum group E gene is developmentally regulated and contains a leucine zipper motif. AB - We have cloned and characterized the Dictyostelium discoideum repE gene, a homolog of the human xeroderma pigmentosum (XP) group E gene which encodes a UV damaged DNA binding protein. The repE gene maps to chromosome 4 and it is the first gene identified in Dictyostelium that is homologous to those involved in nucleotide excision repair and their related XP diseases in humans. The predicted protein encodes a leucine zipper motif. The repE gene is not expressed by mitotically dividing cells, and repE mRNA is first detected during the aggregation phase of development when the cells have ceased dividing and replicating genomic DNA. The mRNA level plateaus by the time the developing cells have entered multicellular aggregates and remains at the same steady-state level for the remainder of development. In addition, we have demonstrated that the level of mRNA is very low in developing cells. These observations suggest that repE may play a regulatory role in development. The data indicate that potential developmental roles for XP-related genes can be profitably studied in this system. PMID- 8710500 TI - Characterization of several kinds of dimer minizyme: simultaneous cleavage at two sites in HIV-1 tat mRNA by dimer minizymes. AB - A minizyme is a hammerhead ribozyme with short oligonucleotide linkers instead of stem-loop II. In a previous study we demonstrated that a minizyme with high activity forms a dimeric structure with a common stem II. Because of their dimeric structure, minizymes are potentially capable of cleaving a substrate at two different sites simultaneously. In order to examine the properties of different kinds of minizyme, we constructed a number of minizymes with short oligonucleotide linkers (2-5 bases) instead of stem-loop II and examined their cleavage activities against HIV-1 tat mRNA. Analyses of melting curves, as well as Arrhenius plots, revealed that, in general, the longer the oligonucleotide linkers, the more stable and more active were the dimer minizymes. All minizymes examined cleaved the target substrate at two sites simultaneously. The activity of the dimer minizyme with a 5 nt linker was higher than that of the parental hammerhead ribozyme because the latter full-sized ribozyme was able to cleave at one site only. PMID- 8710501 TI - Cold shock domain proteins repress transcription from the GM-CSF promoter. AB - The human granulocyte-macrophage colony stimulating factor (GM-CSF) gene promoter binds a sequence-specific single-strand DNA binding protein termed NF-GMb. We previously demonstrated that the NF-GMb binding sites were required for repression of tumor necrosis factor-alpha (TNF-alpha) induction of the proximal GM-CSF promoter sequences in fibroblasts. We now describe the isolation of two different cDNA clones that encode cold shock domain (CSD) proteins with NF-GMb binding characteristics. One is identical to the previously reported CSD protein dbpB and the other is a previously unreported variant of the dbpA CSD factor. This is the first report of CSD factors binding to a cytokine gene. Nuclear NF GMb and expressed CSD proteins have the same binding specificity for the GM-CSF promoter and other CSD binding sites. We present evidence that CSD factors are components of the nuclear NF-GMb complex. We also demonstrate that overexpression of the CSD proteins leads to complete repression of the proximal GM-CSF promoter containing the NF-GMb/CSD binding sites. Surprisingly, we show that CSD overexpression can also directly repress a region of the promoter which apparently lacks NF-GMb/CSD binding sites. NF-GMb/CSD factors may hence be acting by two different mechanisms. We discuss the potential importance of CSD factors in maintaining strict regulation of the GM-CSF gene. PMID- 8710502 TI - Stabilization of triple-stranded oligonucleotide complexes: use of probes containing alternating phosphodiester and stereo-uniform cationic phosphoramidate linkages. AB - Pyrimidine oligonucleotides containing alternating anionic and stereo-uniform cationic N-(dimethylamino-propyl)phosphoramidate linkages [e.g. d(T+T-)7T, d(T+T )2(T+C-)5T and (U'+U'-)7dT, where U' is 2'-O-methyluridine)] are shown to bind to complementary double-stranded DNA segments in 0.1 M NaCl at pH 7 to form triple stranded complexes with the pyrimidine.purine.pyrimidine motif. For each of the sequences investigated, one stereoisomer bound with higher affinity, and the other stereoisomer with lower affinity, than the corresponding all-phosphodiester oligonucleotide. The stereoisomer of d(T+T-)7T that interacted weakly with a dT.dA target in 0.1 M NaCl formed a novel dA.dA.dT triple-stranded complex with poly(dA) or d(Al5C4A15) in 1 M NaCl; in contrast, the stereoisomer that bound strongly to the dT.dA target failed to form a dA.dA.dT triple-stranded complex. PMID- 8710503 TI - Characterization of the interaction between the acidic activation domain of VP16 and the RNA polymerase II initiation factor TFIIB. AB - Contact between a transcriptional activator and one or more components of the RNA polymerase II transcription initiation machinery is generally believed important for activators to function. Several different molecular targets have been suggested for direct contact by herpes simplex virus virion protein VP16, including the general initiation factor TFIIB. In this report we have used several strategies to critically assess this interaction between VP16 and TFIIB. Affinity columns of VP16 bound TFIIB activity from HeLa cell extracts and the binding was reduced by mutations in the activation domain of VP16. In assays of direct binding, VP16 bound recombinant human TFIIB but not Drosophila or yeast TFIIB. Unlike binding from an extract, however, we found that the interaction between VP16 and recombinant human TFIIB was not affected by mutations in VP16 that reduce transactivation. Point mutations within human TFIIB that reduce transactivation by VP16 have been shown to reduce VP16 binding, but we show here that these same mutations critically affect both the important TBP-TFIIB interaction and the ability of TFIIB to support activator-independent basal transcription in vitro. Taken together our results suggest more evidence is needed to support the notion that TFIIB is a functionally important target for the activator VP16. PMID- 8710504 TI - Dual influence of the yeast Cat1p (Snf1p) protein kinase on carbon source dependent transcriptional activation of gluconeogenic genes by the regulatory gene CAT8. AB - The CSRE (carbon source-responsive element) is a sequence motif responsible for the transcriptional activation of gluconeogenic structural genes in Saccharomyces cerevisiae. We have isolated a regulatory gene, DIL1 (derepression of isocitrate lyase, = CAT8), which is specifically required for derepression of CSRE-dependent genes. Expression of CAT8 is carbon source regulated and requires a functional Cat1p (Snf1p) protein kinase. The derepression defect of CAT8 in a cat1 mutant could be suppressed by a mutant Mig1p repressor protein. Derepression of CAT8 also requires a functional HAP2 gene, suggesting a regulatory connection between respiratory and gluconeogenic genes. Carbon source-dependent protein-CSRE complexes detected in a gel retardation analysis with wild-type extracts were absent in cat8 mutant extracts. However, similar experiments with an epitope tagged CAT8 gene product in the presence of tag-specific antibodies gave evidence against a direct binding of Cat8p to the CSRE. A constitutively expressed GAL4 CAT8 fusion gene revealed a carbon source-dependent transcriptional activation of a UAS(GAL)-containing reporter gene. Activation mediated by Cat8p was no longer detectable in a cat1 mutant. Thus, biosynthetic control of CAT8 as well as transcriptional activation by Cat8p requires a functional Cat1p protein kinase. A model proposing CAT8 as a specific activator of a transcription factor(s) binding to the CSRE is discussed. PMID- 8710505 TI - Yeast retrotransposon Ty4: the majority of the rare transcripts lack a U3-R sequence. AB - The retrotransposon Ty4 is found in different yeast strains at only one to three copies per haploid genome. In the present study, we aimed at relating the apparent low transpositional activity of Ty4 to transcriptional features of this element. RT-PCR revealed that Ty4 is transcribed at a very low level, being comparable with that of GAL4. Contrary to other Ty elements, the transcriptional rate of Ty4 is not affected in a sin4 background nor by treatment of cells with alpha factor. From experiments measuring the expression levels in 1acZ fusion constructs, we conclude that Ty4 transcription is repressed by a negative regulating element residing within the LTR, whereas positive cis-acting elements, like those that have been found to mediate expression of Ty1/2 and Ty3, are absent from Ty4. Analysing Ty4 transcript termini by the RACE-PCR method, we found several distinct transcriptional initiation sites. But surprisingly, the majority of the polyadenylated Ty4 transcripts terminate shortly upstream from the 3' LTR boundary, so that these transcripts do not contain a U3-R sequence, which is normally required for obligate strand transfer during DNA synthesis. Thus, the extremely low transcription rate of Ty4 and imperfect Ty4 transcripts are the reason for the low transpositional activity of this element. PMID- 8710506 TI - Exon skipping induced by cold stress in a potato invertase gene transcript. AB - We show that two invertase genes in potato, like most other plant invertase genes, include a very short second exon of 9 bp which encodes the central three amino acids of a motif highly conserved in invertases of diverse origin. This mini-exon is one of the smallest known in plants and pre-mRNA from these genes may be susceptible to alternative splicing, because of a potential requirement for specialized interaction with the splicing machinery to ensure correct processing for the production of a mature mRNA. No evidence of aberrant post transcriptional processing was observed during normal invertase gene expression in potato. The fidelity of post-transcriptional processing of the pre-mRNA from one of the genes was perturbed by cold stress, resulting in the deletion of the mini-exon from some transcripts. This alternative splicing event occurred under cold stress in both leaf and stem, but was not induced by wounding. This adds an example of exon skipping and the induction of alternative processing by cold stress to the small number of transcripts which have been shown to exhibit alternative splicing in plants. The differential sensitivity of post transcriptional processing to cold stress observed for the two transcripts examined will permit further dissection of the nucleotide sequence requirements for their accurate splicing. PMID- 8710507 TI - Complementation of RNA binding site mutations in MS2 coat protein heterodimers. AB - The coat protein of bacteriophage MS2 functions as a symmetric dimer to bind an asymmetric RNA hairpin. This implies the existence of two equivalent RNA binding sites related to one another by a 2-fold symmetry axis. In this view the symmetric binding site defined by mutations conferring the repressor-defective phenotype is a composite picture of these two asymmetric sites. In order to determine whether the RNA ligand interacts with amino acid residues on both subunits of the dimer and in the hope of constructing a functional map of the RNA binding site, we performed heterodimer complementation experiments. Taking advantage of the physical proximity of their N- and C-termini, the two subunits of the dimer were genetically fused, producing a duplicated coat protein which folds normally and allows the construction of the functional equivalent of obligatory heterodimers containing all possible pairwise combinations of the repressor-defective mutations. The restoration of repressor function in certain heterodimers shows that a single RNA molecule interacts with both subunits of the dimer and allows the construction of a functional map of the binding site. PMID- 8710509 TI - A cell-free transcription system for the hyperthermophilic archaeon Pyrococcus furiosus. AB - We describe here the establishment of a cell-free transcription system for the hyperthermophilic Archaeon Pyrococcus furiosus using the cloned glutamate dehydrogenase (gdh) gene as template. The in vitro system that operated up to a temperature of 85 degrees C initiated transcription 23 bp downstream of a TATA box located 45 bp upstream of the translational start codon of gdh mRNA, at the same site as in Pyrococcus cells. Mutational analyses revealed that this TATA box is essential for in vitro initiation of transcription. Pyrococcus transcriptional components were separated into at least two distinct transcription factor activities and RNA polymerase. One of these transcription factors could be functionally replaced by Methanococcus aTFB and Thermococcus TATA bind- ing protein (TBP). Immunochemical analyses demonstrated a structural relationship between Pyrococcus aTFB and Thermococcus TBP. These findings indicate that a TATA box and a TBP are essential components of the Pyrococcus transcriptional machinery. PMID- 8710508 TI - The complete nucleotide sequence of bacteriophage HP1 DNA. AB - The complete nucleotide sequence of the temperate phage HP1 of Haemophilus influenzae was determined. The phage contains a linear, double-stranded genome of 32 355 nt with cohesive termini. Statistical methods were used to identify 41 probable protein coding segments organized into five plausible transcriptional units. Regions encoding proteins involved in recombination, replication, transcriptional control, host cell lysis and phage production were identified. The sizes of proteins in the mature HP1 particle were determined to assist in identifying genes for structural proteins. Similarities between HP1 coding sequences and those in databases, as well as similar gene organizations and control mechanisms, suggest that HP1 is a member of the P2-like phage family, with strong similarities to coliphages P2 and 186 and some similarity to the retronphage Ec67. PMID- 8710510 TI - Purification and characterization of the Pac1 ribonuclease of Schizosaccharomyces pombe. AB - The pac1+ gene of the fission yeast Schizosaccharomyces pombe is essential for viability and its overexpression induces sterility and suppresses mutations in the pat1+ and snm1+ genes. The pac1+ gene encodes a protein that is structurally similar to RNase III from Escherichia coli, but its normal function is unknown. We report here the purification and characterization of the Pac1 protein after overexpression in E. coli. The purified protein is a highly active, double-strand specific endoribonuclease that converts long double-stranded RNAs into short oligonucleotides and also cleaves a small hairpin RNA substrate. The Pac1 RNase is inhibited by a variety of double- and single-stranded polynucleotides, but polycytidylic acid greatly enhances activity and also promotes cleavage specificity. The Pac1 RNase produces 5'-phosphate termini and requires Mg2+; Mn2+ supports activity but causes a loss of cleavage specificity. Optimal activity was obtained at pH 8.5, at low ionic strength, in the presence of a reducing agent. The enzyme is relatively insensitive to N-ethylmaleimide but is strongly inhibited by ethidium bromide and vanadyl ribonucleoside complexes. The properties of the Pac1 RNase support the hypothesis that it is a eukaryotic homolog of RNase III. PMID- 8710511 TI - The Drosophila fork head factor directly controls larval salivary gland-specific expression of the glue protein gene Sgs3. AB - The Drosophila Fork head protein participates in salivary gland formation, since salivary glands are missing in fork head embryos. Here we show that the fork head encoded protein binds to an upstream regulatory region of the larval salivary gland glue protein gene Sgs3. Mobility shift assay in the presence of an anti Fork head antibody demonstrated that the Fork head factor interacts with the TGTTTGC box shown to be involved in tissue-specific Sgs3 expression. Experiments employing a set of oligonucleotide competitors revealed that Fork head binding was prevented by the same single base substitutions that were previously shown to interfere with the TGTTTGC element function in vivo. Furthermore, the anti-Fork head antibody bound to >60 sites of polytene chromosomes, including the puffs of all Sgs genes and Fork head protein was detected in the nuclei of salivary glands of larvae of all examined stages. These data provide experimental evidence for the hypothesis that the protein encoded by the fork head gene is required initially for salivary gland formation and is utilized subsequently in the control of larval genes specifically expressed in this organ. PMID- 8710512 TI - Interactions of the yeast centromere and promoter factor, Cpf1p, with the cytochrome c1 upstream region and functional implications on regulated gene expression. AB - The upstream activation site (UAS) of the cytochrome c1 gene, CYT1, contains sequences for DNA-binding of several transcription factors. Among them are the heme-dependent protein, Hap1p, and the multiprotein complex, Hap2/3/4/5, which mediate transcriptional induction under aerobic conditions and after exhaustion of glucose, respectively. The multiple interactions of nuclear proteins with the UAS region of CYT1 observed in electrophoretic mobility shift experiments are influenced by carbon source and oxygen tension, but are independent of both regulators, Hap1p and Hap2/3/4/5. All protein-DNA complexes obtained are solely due to the association of the centromere and promoter factor 1 (Cpf1p) with the centromere determining element (CDE I)-like motif at the 5' boundary of the UAS(CYT1). This motif overlaps with a consensus sequence for the binding of the general factor Abf1p. Functional analyses after the separate introduction of point mutations into both elements reveal no role for the latter protein and only a minor role for Cpf1p in the regulated expression of CYT1/lacZ chimaeric proteins. However, in cpf1-mutants, induction of CYT1 reaches higher steady state levels and adaptation to aerobic conditions occurs faster than in wild-type. Thus, Cpf1p seems to reduce CYT1 promoter activity under partly inducing conditions, e.g. when only one of the activators, Hap1p or the Hap2 complex, exerts its function. PMID- 8710513 TI - A novel family of TRF (DNA topoisomerase I-related function) genes required for proper nuclear segregation. AB - We recently reported the identification of a gene, TRF4 (for DNA topoisomerase related function), in a screen for mutations that are synthetically lethal with mutations in DNA topoisomerase I (top1). Here we describe the isolation of a second member of the TRF4 gene family, TRF5. Overexpression of TRF5 complements the inviability of top1 trf4 double mutants. The predicted Trf5 protein is 55% identical and 72% similar to Trf4p. As with Trf4p, a region of Trf5p is homologous to the catalytically dispensable N-terminus of Top1p. The TRF4/5 function is essential as trf4 trf5 double mutants are inviable. A trf4 (ts) trf5 double mutant is hypersensitive to the anti-microtubule agent thiabendazole at a semi-permissive temperature, suggesting that TRF4/5 function is required at the time of mitosis. Examination of nuclear morphology in a trf4 (ts) trf5 mutant at a restrictive temperature reveals the presence of many cells undergoing aberrant nuclear division, as well as many anucleate cells, demonstrating that the TRF4/5 function is required for proper mitosis. Database searches reveal the existence of probable Schizosaccharomyces pombe and human homologs of Trf4p, indicating that TRF4 is the canonical member of a gene family that is highly conserved evolutionarily. PMID- 8710514 TI - Pseudouridine synthases: four families of enzymes containing a putative uridine binding motif also conserved in dUTPases and dCTP deaminases. AB - Using a combination of several methods for protein sequence comparison and motif analysis, it is shown that the four recently described pseudouridine syntheses with different specificities belong to four distinct families. Three of these families share two conserved motifs that are likely to be directly involved in catalysis. One of these motifs is detected also in two other families of enzymes that specifically bind uridine, namely deoxycitidine triphosphate deaminases and deoxyuridine triphosphatases. It is proposed that this motif is an essential part of the uridine-binding site. Two of the pseudouridine syntheses, one of which modifies the anticodon arm of tRNAs and the other is predicted to modify a portion of the large ribosomal subunit RNA belonging to the peptidyltransferase center, are encoded in all extensively sequenced genomes, including the 'minimal' genome of Mycoplasma genitalium. These particular RNA modifications and the respective enzymes are likely to be essential for the functioning of any cell. PMID- 8710515 TI - CTF5--a new transcriptional activator of the NFI/CTF family. AB - NFI/CTF is a family of polypeptides involved in stimulating the initiation of adenovirus DNA replication and the activation of transcription driven by RNA polymerase II. Several naturally occurring NFI/CTF variants display distinctive transactivation activities in vivo. To define more precisely the role of the NFI/CTF family in regulating gene expression, we cloned the splice variant CTF5, analyzed transcriptional activation patterns in a yeast transcription assay, and compared it with other CTF proteins. CTF5, which lacks exons 9 and 10 including a CTD-like motif essential for transcriptional activation by full-length CTF1, enhances transcription to a greater extent than CTF1. In addition, CTF5 is even more active than CTF7, which lacks exons 7-9. These findings indicate that CTF proteins formed by differential splicing display a much broader range of transcriptional activities as observed previously. PMID- 8710516 TI - Silencing of the Escherichia coli bgl promoter: effects of template supercoiling and cell extracts on promoter activity in vitro. AB - Regulation of the Escherichia coli bgl promoter involves the catabolite gene activator protein (CAP) and silencer elements that are located upstream and downstream of the promoter and its CAP binding site. The promoter is kept in a repressed state by the silencer elements and other normally active CAP-dependent or -independent promoters are repressed as well when flanked by these elements. To assess the mechanism of promoter repression, single round in vitro transcription was carried out with plasmids bearing either the wild-type bgl promoter or one of two derivatives that escape repression in vivo by different mechanisms: C234 by improving the CAP binding site of the promoter and delta1 by a deletion within the upstream silencer sequence. Repression of the bgl promoter in vitro was shown to depend on template topology and the presence of cellular factors. With negatively supercoiled templates, all three promoters are transcribed to similar extents by purified E. coli RNA polymerase and no CAP dependence is apparent; with relaxed templates, transcription is CAP dependent, but the levels of transcription of the three promoters are comparable. Addition of crude cell extract to the simple transcription system leads to repression of all three promoter alleles in the absence of CAP. Repression of the mutant alleles but not of the wild-type promoter is completely relieved in the presence of the CAP-cAMP complex. The topology of the DNA template is also important in the differential regulation of these promoters. In the case of C234, repression by cell extract is completely relieved by CAP-cAMP on relaxed or negatively supercoiled templates, while complete derepression of delta1 by CAP-cAMP occurs on negatively supercoiled templates only. Repression by cell extract requires the presence of the histone-like protein H-NS. However, H-NS alone does not appear to be sufficient for specific silencing of the wild-type promoter, since repression of all three promoter alleles caused by purified H-NS protein is completely relieved by the CAP-cAMP complex. These data suggest that template topology, H-NS and other cellular factors are involved in the formation of a specific nucleoprotein structure in the bgl promoter-silencer region; the formation of this nucleoprotein structure keeps an otherwise active promoter in an inactive state. PMID- 8710517 TI - Factors affecting fidelity of DNA synthesis during PCR amplification of d(C A)n.d(G-T)n microsatellite repeats. AB - The susceptibility of microsatellite DNA sequences to insertions and deletions in vivo makes them useful for genetic mapping and for detecting genomic instability in tumors. An in vitro manifestation of this instability is the production of undesirable frameshift products during amplification of (dC-dA)n x (dG-dT)n microsatellites in the polymerase chain reaction (PCR). These products differ from the primary product by multiples of 2 nucleotides. We have tested the hypothesis that factors known to affect the fidelity of DNA synthesis may affect (dC-dA)n x (dG-dT)n frameshifting during the PCR. Neither modifications of pH, dNTP concentration, and Mg++ concentration using Amplitaq, nor the use of thermophilic DNA polymerases including UITma, Pfu, Vent and Deep Vent significantly decreased the production of frameshift products during amplification. However, 3'-->5' exonuclease activity in thermophilic DNA polymerases inhibited the accumulation of PCR products containing non-templated 3' terminal nucleotides. Most interestingly, extension temperatures of 37 degrees C during amplification using the thermolabile DNA polymerases Sequenase 1.0, Sequenase 2.0, and 3'-->5' exonuclease-deficient Klenow fragment greatly decreased the production of frameshift products. This method can improve the resolution of heterozygous or mutant (dC-dA)n x (dG-dT)n alleles differing in size by one or two repeat units. PMID- 8710518 TI - Type IIS restriction enzyme footprinting I. Measurement of a triple helix dissociation constant with Eco57I at 25 degrees C. AB - A method is described to measure triple helix dissociation constants by inhibiting the cleavage of a plasmid constructed to contain a target sequence for the triplex forming oligonucleotide (TFO) dT20 by the type IIS restriction enzyme Eco57I. The method relies upon the TFO's ability to block the cleavage reaction by occupying the enzymes cleavage site but not its specific binding sequence. Using this protocol, the dissociation constant for dT20 bound to its target was 0.16 +/- 0.01 microM at 25 degrees C. The accuracy of this experiment was demonstrated by measuring the Kd of an affinity cleavage TFO using Eco57I and Quantitative Affinity Cleavage Titration. Type IIS restriction endonuclease footprinting should be useful for the qualitative and quantitative investigation of ligand-DNA interactions. PMID- 8710519 TI - Sequence and structure determinants of Drosophila Hsp70 mRNA translation: 5'UTR secondary structure specifically inhibits heat shock protein mRNA translation. AB - Preferential translation of Drosophila heat shock protein 70 (Hsp70) mRNA requires only the 5'-untranslated region (5'-UTR). The sequence of this region suggests that it has relatively little secondary structure, which may facilitate efficient protein synthesis initiation. To determine whether minimal 5'-UTR secondary structure is required for preferential translation during heat shock, the effect of introducing stem-loops into the Hsp70 mRNA 5'-UTR was measured. Stem-loops of -11 kcal/mol abolished translation during heat shock, but did not reduce translation in non-heat shocked cells. A -22 kcal/mol stem-loop was required to comparably inhibit translation during growth at normal temperatures. To investigate whether specific sequence elements are also required for efficient preferential translation, deletion and mutation analyses were conducted in a truncated Hsp70 5'-UTR containing only the cap-proximal and AUG-proximal segments. Linker-scanner mutations in the cap-proximal segment (+1 to +37) did not impair translation. Re-ordering the segments reduced mRNA translational efficiency by 50%. Deleting the AUG-proximal segment severely inhibited translation. A 5-extension of the full-length leader specifically impaired heat shock translation. These results indicate that heat shock reduces the capacity to unwind 5-UTR secondary structure, allowing only mRNAs with minimal 5'-UTR secondary structure to be efficiently translated. A function for specific sequences is also suggested. PMID- 8710520 TI - Selective cloning of a defined number of tandem DNA repeats in Escherichia coli. PMID- 8710521 TI - A strong ubiquitous promoter-enhancer for development and aging of Drosophila melanogaster. PMID- 8710522 TI - Differential screening of gene expression difference enriched by differential display. PMID- 8710523 TI - The use of alpha-DNA as an internal standard in the detection and quantitation of DNA damage in specific genes using Southern blotting. PMID- 8710524 TI - Hetero-stagger cloning: efficient and rapid cloning of PCR products. PMID- 8710525 TI - 'Long distance sequencer' method; a novel strategy for large DNA sequencing projects. PMID- 8710527 TI - [Nuclear medical breast diagnosis--scintigraphy using 99mTc-sestamibi]. PMID- 8710526 TI - PET imaging of recurrent medullary thyroid cancer. AB - Medullary carcinoma of the thyroid gland is a rare tumor. Its prognosis is mainly linked to surgery, because there is no valid alternative therapy to improve patients outcome. In this report, we discuss the recurrence of such a tumor in a 64-year-old female, focusing on magnetic resonance imaging and position emission tomography evaluation of this tumor. PMID- 8710528 TI - [Differential diagnosis of childhood osteomyelitis--classification according to scintigraphic, radiologic and magnetic resonance tomographic characteristics]. AB - We present a retrospective study on children with the final diagnosis osteomyelitis, who have been examined in Tubingen from 1985 to 1991. The different types of infantile osteomyelitis were classified due to the causative organism and findings in 3-phase scintigraphy and X-ray films. For the chronic type of osteomyelitis the study was extended to the years from 1979 to 1991 and the results of an earlier report were included. We worked up 17 cases of acute/peracute osteomyelitis, including 5 cases of early infancy, 2 cases of tuberculosis, 2 Brodie's abscesses, 5 plasmacellular types, 2 cases of primary chronic multifocal osteomyelitis (PCMO), and 5 cases of unspecific chronic osteomyelitis. All cases were examined with scintigraphy, X-ray films and in part with magnetic resonance tomographic imaging. In 23 cases scintigrams and X-ray films were performed in the follow-up. We show the importance of scintigraphy for the early detection and localisation of osteomyelitis, the importance of findings on X-ray films for the specific diagnosis of osteomyelitis, and the importance of magnetic resonance tomography for high-resolution detection of the expansion of osteomyelitis. PMID- 8710529 TI - [201Tl myocardial SPECT. First experiences with a simultaneous transmission emission acquisition protocol for patient-specific attenuated correction]. AB - AIM: In this study our first clinical experiences with simultaneous transmission and emission acquisition in 201 TI myocardial SPECT (T/E-SPECT) are discussed. METHODS: The non-uniform attenuation (AK) was carried out with a triple-head camera (PRISM 3000, Picker Inc.) correction equipped with fanbeam collimators. A line source of 750 MBq 99mTc was used to construct the transmission profile. Prior to investigation patients got 80-120 MBq 201TI-chloride intravenously injected. RESULTS: The study comprises the evaluation of 40 patients, derived from the clinical routine. The investigation followed an usual one day protocol. Our results using T/E-SPECT reveal an almost equilibrated activity distribution between anterior and posterior myocardial wall. CONCLUSION: For this reason it is to be expected that T/E-SPECT provides more reliable information about the posterior myocardial wall, than the usual SPECT technique without attenuation correction. PMID- 8710530 TI - [75-Se-labeled bile acid analog absorption in various gastrointestinal diseases using a whole body counter]. AB - AIM: It is possible to detect disturbances of bile acid absorption using a whole body counter after administration of Se-75 labelled bile acid analogues. We scrutinized the benefit of a modification of the test method. METHODS: We investigated 77 patients with different forms of a gastrointestinal disease. After application of Se 75 homotaurocholic acid we measured patient-activity up to 7 days later including whole-body profile scans in the first 6 h. RESULTS: The fractional retention after 7 days was between 20 and 67%. In cases of impaired absorption it was below 12%. Patients with liver diseases and after cholecystectomy (without bile acid resorption disturbance) showed normal values. Patients with Crohn's disease of the ileum or with intestinal ileac by-pass or with colestyramine treatment or with disturbance of vitamin B12-absorption or with cystic fibrosis showed a disturbance of bile acid absorption. The normal whole-body half-life was more than 2.8 days. The 24 and 72 h values were 62 and 31% in cases with normal absorption. Smaller values are signs of bile acid malabsorption. Impulse rates measured with the whole body counter are of an order of magnitude that allows to reduce the usually administered dose of 37 kBq to 9.25 kBq. CONCLUSION: This is an efficient method to detect disturbances of bile acid absorption. The usually administered activity of 37 kBq can be reduced to 9.25 kBq. PMID- 8710531 TI - [Methodical evaluation of immunoluminometric determination of thyroid peroxidase in serum]. AB - With the identification of thyroid peroxidase (TPO) as the specific autoantigen in autoimmune diseases of the thyroid, the development of a commercial assay for detection of TPO in human serum became possible. The diagnostic value of this TPO assay was evaluated in 194 patients with various thyroidal diseases. The assay appeared to be easily affected by specific and/or unspecific interferences such as TPO-auto-antibodies in the patient's blood samples. To analyze these effectors every sample was checked in a parallel recovery test. In most of the cases with elevated anti-TPO levels an exact determination of TPO could not be estimated correctly. Whenever a correct measurement of TPO was possible, to none of the different examined groups of thyroid diseases a correlation of TPO-levels could be demonstrated. Moreover, the value of TPO determination as a tool in the follow up of differentiated thyroid carcinoma was not provable. For the time being our studies do not suggest TPO measurements being helpful in thyroidal diagnosis. PMID- 8710532 TI - Cor triatriatum dextrum--an unusual variant of triatrial heart. AB - The present report describes a rare case of triatrial heart, cor triatriatum dextrum in a 31-year-old woman in whom the anomaly was incidentally detected by radionuclide angiocardiography. Subsequent assessment with two-dimensional echocardiography and right heart catheterization confirmed the presence of a membrane subdividing the right atrium into two distinct chambers. To the best of our knowledge, this is the only case in which this cardiac anomaly was demonstrated by radionuclide technique. PMID- 8710533 TI - Professional development. Breast cancer: knowledge for practice (continuing education credit). PMID- 8710534 TI - Righting wrongs. Interview by Adam Legge. PMID- 8710535 TI - When a fatal error is made worse. PMID- 8710536 TI - Satisfied customers. PMID- 8710537 TI - The establishment of a gynaecology assessment unit. AB - The team of the gynaecological surgery ward at Watford General Hospital have established a gynaecological assessment unit run by a nurse or midwife practitioner. In this paper a member of the management team outlines the scheme. PMID- 8710538 TI - Women's health. Help for substance misusers. AB - Evidence suggests that heavy drinking among women is increasing but few services have managed to target them successfully. This paper describes the work of a weekly drop-in session for women with alcohol and drug problems run by Riverside Mental Health NHS Trust, London, which attracts up to 23 clients at each session. PMID- 8710539 TI - Making sense of the process and effects of ECT. AB - An estimated 20,000 people are treated with electroconvulsive therapy every year. This paper provides an overview of what is involved and some of the issues arising from this controversial treatment. PMID- 8710540 TI - Stress and clinical supervision in mental health care. AB - Clients with severe mental health problems are a challenge to mental health services. Working with such clients can be stressful and may ultimately lead to emotional exhaustion. This paper looks at the introduction of clinical supervision in a continuing care team, aiming to provide support and advice to community nursing staff. PMID- 8710541 TI - Support for pregnant women who wish to stop smoking. AB - More than a quarter of pregnant women continue to smoke. This paper looks at how health professionals can support clients who wish to give up tobacco. It also examines the ways in which nurses and midwives can help pregnant women overcome barriers to cessation. PMID- 8710542 TI - Long-term care. Small change. PMID- 8710543 TI - Learning disabilities. Sensory growth. PMID- 8710544 TI - The last frontier. PMID- 8710545 TI - Complementary medicine. Opening up new relief routes. PMID- 8710546 TI - Pulp fiction. PMID- 8710547 TI - Infection control. The toy trap uncovered. PMID- 8710548 TI - Infection control. An overhaul of home loans. PMID- 8710549 TI - Professional development. Breast cancer: revision notes (continuing education credit). PMID- 8710550 TI - Safe sex advice for mental health patients. PMID- 8710551 TI - Open all hours. PMID- 8710552 TI - Old diseases die hard. PMID- 8710553 TI - Eastern affairs. PMID- 8710554 TI - Public health: global action. PMID- 8710556 TI - Issues for clinical leaders in nursing development units. AB - This study explores the effect of grants on the clinical leaders of nursing development units (NDUs). A qualitative exploratory study was carried out by The King's Fund Centre using a semi-structured interview schedule. Through the interviews clinical leaders were able to explain how they felt about the responsibility of leading a funded NDU. The information gathered was examined for common themes and categories which revealed the complexity of effects on clinical leaders. PMID- 8710555 TI - Public health: unequal approach. PMID- 8710557 TI - Laboratory measurement of sodium and potassium. AB - This paper continues our series on pathology testing. The urea and electrolyte (U&E) blood test, which includes the measurement of sodium and potassium concentration in blood plasma, is the most frequently requested chemical test in clinical practice. The precautions to be taken when sampling blood for this test are described. This is followed by a brief review of conditions associated with abnormalities in serum sodium and potassium concentrations and the effects of these changes. It is particularly important that the signs and symptoms of changes in plasma potassium concentration particularly are recognised and treated quickly, as these are potentially life-threatening. PMID- 8710558 TI - The incidence of malnutrition in hospitalised patients. AB - This paper is the third in our literature review series and looks at the subject of nutrition. In 1859 Florence Nightingale suggested that thousands of patients were 'annually starved in the midst of plenty, from want of attention to the ways which alone make it possible for them to take food'. Over 130 years later it is still being suggested that nutrition remains a neglected area of patient care. It is sobering to realise that little has changed in this time. This paper provides an overview of malnutrition in hospitals and investigates the factors contributing to the high incidence of nutritional depletion which, even today, is so common in hospital patients. PMID- 8710560 TI - Patient experience: an alien place. PMID- 8710559 TI - The case for breaking through ageism in mental health care. AB - This paper critically examines the recent policy in mental health nursing of refocusing care on people with severe and enduring mental health problems. It is argued that older people with severe and enduring mental illnesses such as dementia are excluded from this policy and that this amounts to a clear case of ageism. Various ways of addressing the problem of ageism in recent policy changes are explored. PMID- 8710561 TI - Leadership challenge. PMID- 8710562 TI - NT Midwife. Rediscovery of the lost art. PMID- 8710563 TI - NT Midwife. Is breast-feeding 'natural'?--A health visitor's view. PMID- 8710564 TI - NT Midwife. Against all odds. PMID- 8710565 TI - NT Midwife. The Stepford midwives. PMID- 8710566 TI - Professional development. Termination of pregnancy: the role of the nurse (continuing education credit). PMID- 8710567 TI - Safety at work. A healthy decision. Interview by Michael Day. PMID- 8710568 TI - Safety at work. Counterstrike. PMID- 8710569 TI - Inadmissible evidence? PMID- 8710570 TI - Campaign aims to change the culture on manual lifting. AB - The RCN this month launches a long-term campaign attempting to reduce injuries to nurses by abolishing almost all lifting by nurses. The campaign will be accompanied by new RCN literature and legal advice. Both the RCN and the UKCC anticipate that community nursing may be a problematic area for the introduction of non-lifting policies. The implementation of a minimum-lifting policy at one NHS trust is studied, and the early results reported. PMID- 8710571 TI - Lifting and handling. Strategies for altering practice. AB - The elderly services directorate at the Ipswich Hospital NHS Trust has already embarked on changing nurses' patient-handling practice in response to EC regulations. This paper describes how problems hindered the programme's initial success and explores how new strategies have been implemented to overcome them. PMID- 8710572 TI - Systems of life. Abnormalities of puberty. PMID- 8710573 TI - Finding, reading and documenting literature. AB - Before embarking on an evaluation project it is important to be familiar with the relevant literature. Critical reading of the literature is essential in helping the investigator refine the questions and design of his or her study. The retrieval and use of available literature for use in a study proposal is outlined here. PMID- 8710574 TI - Guidance on latex sensitisation for the health service. AB - Following NT's successful Live Issues series on the problem of latex allergies, this paper provides the Medical Devices Agency (MDA) view of the situation in the health service and describes the MDA's new guidance on the subject. PMID- 8710575 TI - Sexual abuse and motherhood. PMID- 8710576 TI - Nursing overseas. A lesson in litigation. PMID- 8710577 TI - Euthanasia. Deepening dilemma. PMID- 8710578 TI - Pressure sores. Damage limitation. PMID- 8710579 TI - Speaking terms. PMID- 8710580 TI - Continence. Outreach for prevention. PMID- 8710581 TI - Continence. Delivering the best service. PMID- 8710583 TI - Continence. Working together. PMID- 8710582 TI - Continence. Comprehensive care to order. PMID- 8710584 TI - Professional development. Termination of pregnancy: revision notes (continuing education credit). PMID- 8710585 TI - Bitter rain. PMID- 8710586 TI - Supervision or coercion? PMID- 8710587 TI - Offensive conduct. PMID- 8710588 TI - PREP. A year on the learning curve. PMID- 8710589 TI - PREP. Not up to standard. PMID- 8710590 TI - A public health nursing post: the tools for getting started. AB - This paper looks at the progress of an estate-based public health post in the first six months of its existence. It examines some of the tools used in this critical period, rather than describing the estate in detail. The activities of the post are reviewed in terms of information-gathering, action networking and analysis of needs and strengths. The paper describes a framework which seeks to reconcile the demand for community action with the development of mainstream services. The challenges of making wider use of the post's activities in the traditional primary health care team are explored. The development of health-gain indicators for the post is also highlighted. PMID- 8710591 TI - Use of tumour markers in cancer diagnosis and treatment. AB - This paper examines the laboratory identification of tumour markers--substances present in the blood and urine that are released by tumour cells or by the patient in response to a tumour. A general consideration of what constitutes a useful tumour marker is followed by a description of some specific tumour markers currently used in clinical practice. Reference is made to the potential impact of recent cancer research on initiatives for early cancer detection by testing blood. PMID- 8710592 TI - Prisoners with diabetes: do they receive appropriate care? AB - This paper reports on a survey of the care delivered to people with diabetes in a small number of prisons, focusing on diets and the availability of diabetic clinics. PMID- 8710593 TI - A therapeutic approach to bereavement counselling. AB - Nurses are increasingly caring for bereaved people therefore a basic knowledge of the grieving process and appropriate interventions are essential. Research has shown that factors surrounding death can predispose bereaved people to experiencing abnormal and complicated grief processes. One such factor is a sudden bereavement. One third of all deaths in hospitals occurs suddenly and unexpectedly. In this article, nursing interventions are explored and discussed along with implications for nursing theory, practice, management and education. PMID- 8710594 TI - Taken to task. PMID- 8710595 TI - Wound cleansing: principles and practice. PMID- 8710596 TI - Wise up to the real world. PMID- 8710597 TI - Ageing matters. Coordinators of care. PMID- 8710598 TI - Ageing matters. Homing in on drug rounds. PMID- 8710600 TI - On duty. Interview by Antonia Owen. PMID- 8710601 TI - Going straight to a new start in life. PMID- 8710599 TI - Careful talk saves lives. PMID- 8710602 TI - Learning from our mistakes. PMID- 8710604 TI - New deal for nurses now. The government must show its commitment to improving nurses' pay and conditions before morale slumps even further. PMID- 8710603 TI - The route to effective nurse-patient communication. AB - Communication is a central part of nursing practice. The importance of good communication in the delivery of effective and appropriate nursing care has been well demonstrated by research and is reflected in policy documents. However, despite the emphasis on teaching communication skills over the past two decades, good communication is often curtailed by structural factors. These include the nature of ward organisation, remote management styles, and an overwhelming emphasis on increased patient throughput. Increased day surgery together with the greater use of bank and agency staff place further limits on nurses' ability to develop a substantial rapport with patients. It is concluded that adequate time and support are needed to enable nurses to communicate therapeutically with patients in order to achieve care that is effective and responsive to their needs. PMID- 8710605 TI - Findings of a national survey of acute pain services. AB - This paper presents the results of a national acute pain survey carried out in 1994. The survey aimed to establish the current status of acute pain management after major surgery in hospitals in the UK and to pinpoint those hospitals with acute pain services. The survey identified that in 1994 recommendations concerning the structure and provision of acute pain services had been partially met in fewer than half the 221 hospitals that responded. The majority of acute pain services consisted of medical and nursing staff; few included psychological or pharmaceutical expertise. The main reason cited for the underdevelopment of services was lack of funding. PMID- 8710606 TI - Percutaneous endoscopic gastrostomy: a review. AB - This paper continues our series of literature reviews and describes the technique of percutaneous gastrostomy. The aftercare and complications of this method of feeding are also considered, and the prevention of infection and psychological aspects of nutritional support discussed. PMID- 8710607 TI - Future treatment options for haematological malignancies. AB - This paper is the last in a series looking at haematological malignancies. It pinpoints where we are now in the treatment of leukaemias before discussing current and future developments. PMID- 8710608 TI - The local pay lottery. PMID- 8710610 TI - Coeliac disease: maintaining a gluten-free diet. PMID- 8710609 TI - Birthday credits due. PMID- 8710611 TI - Too hot to handle. PMID- 8710612 TI - Taking an active role. PMID- 8710613 TI - At the heart of the action. PMID- 8710614 TI - How patients see symptoms. PMID- 8710615 TI - Psychological pain and care. PMID- 8710616 TI - Long-term prospects. PMID- 8710617 TI - Professional development. Electrocardiography. Knowledge for practice. PMID- 8710618 TI - Words fail us. PMID- 8710619 TI - Time to talk. PMID- 8710620 TI - Divided we stand. PMID- 8710621 TI - Baby scoring system misses the point. PMID- 8710622 TI - Learning disabilities: sold short. PMID- 8710623 TI - Difficulties of a double role. PMID- 8710624 TI - Learning disabilities. Expecting a choice. PMID- 8710625 TI - Learning disabilities. Qualifying existence. PMID- 8710626 TI - Can timed appointments for community staff improve care? AB - A new Patient's Charter standard relating to timed appointments for community nurses was introduced in 1995. To date, research considering the implications of this standard has been limited, particularly in terms of rural communities. This paper describes a study which explores the issues surrounding the implementation of the standard in rural Shropshire. It measured the success rate with which the standard was met and considered the perceptions of nurses and clients. The paper concludes that if a 100% success rate in meeting the standard is to be achieved significant changes will need to take place in the organisation of community nursing services. PMID- 8710627 TI - Making sense of bovine spongiform encephalopathy. AB - The possibility of animals infected with bovine spongiform encephalopathy (BSE) causing Creutzfeldt-Jakob disease (CJD) in humans has caused public alarm resulting in the banning of the export of British beef. This paper explores the current understanding of BSE and its relationship to CJD. Although there are similarities between the diseases, there is no clear evidence of a causal link between the two. PMID- 8710628 TI - Reasons for pre-hospital delay in reporting chest pain. AB - This paper looks at why there are still delays between onset of chest pain in myocardial infarction and administration of thrombolytic therapy. It looks at the research into these delays and sets out a number of reasons for them. It also recommends courses of action and further studies to address the problem. PMID- 8710629 TI - Use of eye movement to reduce stress after trauma. AB - In 1987 an accidental discovery revealed an association between certain eye movements and reduced levels of distress resulting from traumatic memories. The result was a new psychological intervention, eye movement desensitisation and reprocessing (EMDR). The treatment consists of generating rapid and rhythmic eye movements while simultaneously holding traumatic images, thoughts and emotions in the active memory. This paper describes the experiences of one psychotherapist in using EMDR to treat people with post-traumatic stress disorder. Six case studies illustrate aspects of this complex treatment. PMID- 8710630 TI - Contact points. PMID- 8710631 TI - Travel abroad: immunisation advice. PMID- 8710632 TI - Controlling interests. PMID- 8710633 TI - Precepts of care. PMID- 8710634 TI - Help where needed. PMID- 8710636 TI - The chieftain. Interview by Janet Snell. PMID- 8710635 TI - Professional development. Electrocardiography: the role of the nurse (continuing education credit). PMID- 8710637 TI - Good cause for complaint. PMID- 8710638 TI - Shared learning. PMID- 8710639 TI - Postnatal depression: how can the distress be alleviated? AB - This paper discusses the incidence, manifestations and predisposing factors of postnatal depression. The different theories about its underlying causes are reviewed, together with the associated evidence. The implications of this for the practice of community nurses and midwives are then considered in terms of the prevention, detection and treatment of the condition. PMID- 8710640 TI - A personal account of postnatal depression. PMID- 8710641 TI - Clinical evaluation: deciding what questions to ask. AB - Preparation and planning ahead are all-important in carrying out an evaluative study. Systematic evaluation provides evidence for the justification of a given treatment or intervention. In this third article looking at clinical evaluation, the process of posing a question which lends itself to study is examined. Questioning, it is argued here, can be seen as having three progressively complex levels. These can uncover: first, whether an effect has taken place; second, what may have caused the effect; and third, how the effect might be improved. Previous articles in this series appeared in the March 13 and April 10 issues of Nursing Times. PMID- 8710642 TI - Systems of life. Female sexual function. PMID- 8710643 TI - Critical of care. PMID- 8710644 TI - Cardiac distress. PMID- 8710645 TI - Nursing abroad. It's a jungle out there. PMID- 8710646 TI - Infection control. Keeping MRSA in perspective. PMID- 8710647 TI - Delegates protest at 'malicious' cuts in HIV funding. PMID- 8710648 TI - Infection control. Putting up a united front. PMID- 8710649 TI - Infection control. Time to freshen up. PMID- 8710650 TI - An age-old entitlement. PMID- 8710651 TI - Right to life. PMID- 8710652 TI - How care in the community has affected older people. AB - This paper gives an overview of the current shortfalls in the nursing care delivered to older people since the introduction of the NHS and Community Care Act. Descriptions of recent reports and studies are included alongside views of those involved in protecting the interests of older people PMID- 8710653 TI - The single care home. AB - This paper discusses the importance of nursing in the long-term care setting and the issues arising from the possible introduction of a single registration care home. The current situation is examined and the implications for patients are highlighted. PMID- 8710654 TI - Use of remodelling in the treatment of sex offenders. AB - This grounded theory study explored the perceptions of a group of sex offenders receiving treatment in order to generate an explanatory theory of the sexual abuse treatment process. The subjects were male sex offenders who were in a community sexual abuse treatment programme; a number had either dropped out of the programmed or had graduated from it. Constant comparative analysis was used to collect and analyse the data concurrently. The results suggest that adult male sex offenders undergo a remodelling process whereby they face up to the discovery of their abuse and go through the treatment process. Using remodelling as a metaphor offers a concrete, visual way to discuss the treatment process and expectations with individual offenders in order to help them avoid reoffending in the future. PMID- 8710655 TI - Systematic search offers a sound evidence base. AB - This is the first paper is a series that will look at literature reviews as a method of using information to improve practice. This paper explains why we need to use literature review, how we can gain access to the information, how to do the review and, finally, how to write it up. The following three papers will use an example of a task in nursing practice and apply the method of literature review to it. The second paper will be published in the February 28 issue of Nursing Times and will look at oral care. PMID- 8710656 TI - Therapy with attitude. PMID- 8710657 TI - Who was Meniere? PMID- 8710658 TI - A condom's tale. PMID- 8710659 TI - Injury time. PMID- 8710660 TI - Stoma: treatment and appliances. PMID- 8710661 TI - Degrees of nonsense. PMID- 8710662 TI - A safer passage. PMID- 8710663 TI - A harmonious atmosphere. PMID- 8710664 TI - Hidden danger. PMID- 8710665 TI - Two-tier treatment: why pick on us? PMID- 8710666 TI - Totting-up tactics. PMID- 8710667 TI - Stoma care. Choice cuts. PMID- 8710668 TI - Stoma care. Self-care at home. PMID- 8710669 TI - Audit of nutrition screening in patients with acute illness. AB - This paper describes a nine-month comparative audit project looking at the assessment and management of patients' nutritional status. The findings show a significant improvement in nutritional management following a staff training programme. PMID- 8710670 TI - Key issues in the introduction of nurse endoscopy. AB - The inspection technique of gastrointestinal endoscopy is becoming increasingly popular and services in this area are expanding. Conditions which could only be treated by surgery can now be done endoscopically and this requires suitably trained medical staff. Consequently, there are demands from the medical and nursing professions to develop the role of the nurse endoscopist. This paper raises issues of concern to nurses who wish to undertake diagnostic endoscopy and suggests how the necessary skills and knowledge to gain competence can be obtained. PMID- 8710671 TI - Measurement of aspirin and paracetamol metabolites. AB - Clinical laboratories are frequently asked to measure the concentration of drugs in blood and urine. Of the many thousands of drugs prescribed, however, the measurement of only a relatively small number provides useful clinical information. There are three main reasons for measuring drugs: to test patient compliance, to ensure that dosage is high enough to have therapeutic effect but sufficiently low to avoid toxicity and, finally, to identify drugs taken during deliberate or accidental overdose. This article is concerned with the last of these. PMID- 8710672 TI - Multidisciplinary approach to patient communication. AB - The nursing team in a 28-bed rehabilitation ward for older people at South Cleveland Hospital recognised a need to improve communications between patients, carers, relatives and members of the medical team. They developed a model for a patients, families and carers clinic to address these issues. This was then tested in the ward over a three-month period, a pilot that continued after the ward had been modified to a 20-bed stroke unit. PMID- 8710673 TI - An approach to dealing with stress in carers. AB - This article examines the effect of stress on carers of children with learning difficulties. One family's problems, in relation to their psychological well being, are related to relevant ideas, concepts and theories associated with stress and coping. Implications for future practice are also discussed. PMID- 8710674 TI - From pillar to post. PMID- 8710675 TI - International perspectives. The unseen menace. PMID- 8710676 TI - A new way of life. PMID- 8710677 TI - Who was Charcot? PMID- 8710678 TI - Handling aids for nurses and carers. PMID- 8710679 TI - Appliance of science. PMID- 8710680 TI - Dietary cholesterol inhibits the development of aberrant crypt foci in the colon. AB - We evaluated the effect of dietary cholesterol and oxidized cholesterol on the promotion of aberrant crypt foci (ACF), which are putative precancerous lesions in the colon. Sixty female C57BL/6J mice were given four weekly injections (ip) of azoxymethane (AOM) then fed either a control AIN-76 diet or the control diet supplemented with 0.3% cholesterol or 0.3% oxidized cholesterol for 100 days. The oxidized cholesterol was prepared by heating cholesterol at 110 degrees C for 48 hours. Gas chromatographic analysis of the oxidized cholesterol showed that 96% of the cholesterol was unchanged and less than 2% of the cholesterol was oxidized. The remaining 2% impurities were unidentified and present in both the cholesterol and heated cholesterol. The number of ACF in the group fed cholesterol was significantly lower than the control group (7.9 +/- 1.0 vs. 12.5 +/- 1.2, p < 0.01). The number of ACF in the group fed oxidized cholesterol (10.1 +/- 1.1) was not different from the control or cholesterol groups. The size of the ACF (no. of crypts per focus) did not differ between the three dietary groups. Serum low-density lipoprotein (LDL) cholesterol was greater in the cholesterol-fed group than the control group (40.5 +/- 4.6 vs. 24.3 +/- 3.6 mg/dl, p < 0.05). LDL cholesterol from the animals fed oxidized cholesterol (37.7 +/- 4.7 mg/dl) was not different from the control or cholesterol-fed animals. Total and high-density lipoprotein (HDL) cholesterol did not differ between the groups. The results show that dietary cholesterol significantly inhibits the promotion of ACF in the colon. The elevated LDL cholesterol may inhibit de novo cholesterol synthesis in the preneoplastic colonic epithelial cells, thereby inhibiting DNA synthesis and cell proliferation. PMID- 8710681 TI - Effects of alpha-tocopherol and beta-carotene on hepatic lipid peroxidation and blood lipids in rats with dietary iron overload. AB - The ability of dietary antioxidants to reduce lipid peroxidation induced by iron overload was examined in weanling male Sprague-Dawley rats. Animals were fed ad libitum a modified AIN-76A diet (control) or control diet with 0.5% alpha tocopherol acid succinate, 0.5% crystalline trans-beta-carotene, or 0.5% alpha tocopherol acid succinate + 0.5% trans-beta-carotene for four weeks. In the following four-week period, the animals received the above diets with 10,000 micrograms Fe/g; a control group continued to receive 35 micrograms Fe/g, and a high-iron group received 10,000 micrograms Fe/g with no antioxidants. After four weeks of dietary supplementation with alpha-tocopherol. Beta-carotene or alpha tocopherol + beta-carotene, liver concentrations of alpha-tocopherol and beta carotene increased significantly (p < 0.001). Liver lipid peroxidation, measured by the lipid-conjugated diene assay, increased significantly from 0.012 mumol/mg of lipid in the controls to 0.021 mumol/mg of lipid in animals receiving the high iron diet. However, lipid peroxidation was significantly reduced in all animals fed the antioxidants, with the group fed alpha-tocopherol + beta-carotene having a lower level than the high-iron group. Total serum cholesterol was elevated in animals fed a high-iron diet and in animals fed the high-iron diet with alpha tocopherol. In contrast, total serum cholesterol levels in the two groups of animals receiving the diets containing high iron with beta-carotene alone or high iron with beta-carotene + alpha-tocopherol were significantly reduced to the level of the control group. High-density lipoprotein cholesterol also decreased to baseline in the animals receiving beta-carotene alone. Modulation of lipid peroxidation by alpha-tocopherol or beta-carotene may be an important mechanism for reducing oxidative stress. PMID- 8710682 TI - Regional distribution of carcinogen-induced colonic neoplasia in the rat. AB - Carcinogen induction of neoplasms in rodent colon has been used as a model for human colon cancer development and for evaluating chemopreventive regimens. We studied the regional distribution of small and large intestinal tumors in 229 rats given azoxymethane (AOM) once weekly for two weeks (15 mg/kg sc). The AOM regimen induced 63% more tumors in distal (DC) than in proximal colon (PC), although tumor volume was greater in PC. A high-fat (23% corn oil) diet increased tumors in PC and DC (p < 0.01). Caloric restriction of 10-30% of the ad libitum diet progressively reduced DC tumor formation but did not alter PC tumors. Tumor volume was unaffected by either regimen. Small intestinal tumors were concentrated in the proximal 15 cm of the intestine and were unaffected by dietary manipulation. This AOM model of colon tumor formation approximates human colon cancer distribution and is an appropriate model for rodent chemopreventive studies. PMID- 8710683 TI - Modulation of murine EL-4 thymic lymphoma cell proliferation and cytokine production by vitamin E succinate. AB - RRR-alpha-tocopheryl succinate (VES) was studied for effects on murine EL-4 cell proliferation and production of interleukin-2 (IL-2) and transforming growth factor-beta (TGF-beta). VES was biphasic in its actions: 0.1 microgram/ml enhanced EL-4 cell proliferation, whereas 10-20 microgram/ml inhibited cellular proliferation. Cell-conditioned media (CM) from EL-4 cells treated with 0.2 ng/ml phorbol myristate acetate (PMA) + 0.1 microgram/ml VES contained increased amounts of IL-2, as determined by the murine cytotoxic T cell IL-2-dependent CTLL 2 bioassay. VES at 0.1 microgram/ml or 0.1 microgram/ml VES + 0.2 ng/ml PMA induced the expression of IL-2 mRNA by EL-4 cells three to nine hours after treatment. CM from EL-4 cells treated with VES at 10-20 microgram/ml exhibited potent antiproliferative activity when tested in the TGF-beta-responsive mink lung cell (Mv1Lu) bioassay and showed reduced inhibitory effects when tested on TGF-beta receptor-negative mink lung (DRA-27) cells. CM from control-treated EL-4 cells exhibited no antiproliferative activity. The VES-induced antiproliferative activity was characterized as TGF-beta by neutralization analyses and immunoprecipitation of metabolically labeled proteins with TGF-beta-specific reagents. VES treatment of EL-4 cells had no effect on TGF-beta 1 mRNA expression while downregulating TGF-beta 3 mRNA expression. In summary, these studies showed that 0.1 microgram/ml VES enhanced cellular proliferation, in part, via increased IL-2 production, whereas 10-20 micrograms/ml VES inhibited cellular proliferation, in part, via the secretion of biologically active TGF-beta. PMID- 8710684 TI - The effect of dietary fat on the promotion of mammary and colon cancer in a dual organ rat carcinogenesis model. AB - Female Sprague-Dawley rats (n = 270) were randomly divided into nine subgroups (30 rats/group) and fed diets with fat contents ranging from 5% to 45% of total energy, in which fat was isocalorically substituted for carbohydrates. At Week 3, the rats were given a single injection of methylnitrosourea (MNU, 50 mg/kg body wt). At Weeks 4 and 5, rats were given an injection of azoxymethane (AOM, 15 mg/kg body wt sc). The rats were maintained on their respective experimental diets until they were sacrificed. Nine weeks after the injection of MNU, 10 rats from each group were killed and their mammary tissue was examined for tumors and their colons for aberrant crypt foci (ACF) formation. The results showed no significant increase in the number of colonic ACF or mammary tumors at 11 weeks among individual groups consuming 5-25% energy from fat or among groups consuming 30-45% energy from fat. Data analysis indicated that the effects of the two diets (5-25% energy from fat and 30-45% energy from fat) on ACF formation as well mammary tumor development were statistically different. Continuation of the diets for 32 weeks led to the development of colonic and mammary tumors. Analysis of the combined group tumor incidence data clearly showed that the colonic and mammary tumor incidence in groups fed 30-45% of energy from fat was significantly higher than that observed among groups fed 5-25% of energy from fat. Linear regression analysis of the colonic ACF at 11 weeks, colonic tumor incidence at 32 weeks, and mammary tumor incidence at 11 and 32 weeks showed good linear correlation, demonstrating the potential value of ACF and mammary tumors at 11 weeks for evaluating the carcinogenic risk associated with different diets. The main conclusion of the experiment is that the incidence of colon and mammary cancers increased rapidly when the dietary levels of fat were increased from 15% to 30% of calories and that there was no significant influence on the colon and mammary cancer risk beyond 30% of calories from fat. PMID- 8710685 TI - Effects of two dietary fat levels and four dietary linoleic acid levels on mammary tumor development in Balb/c-MMTV mice under ad libitum feeding conditions. AB - The relationship between dietary fat intake (level and type) and the development of breast cancer in humans is a matter of concern in Western society. A high fat intake is associated with a greater mammary cancer risk in humans and in animal models. Higher intake of polyunsaturated fatty acids in humans shows little or no association with mammary tumor development in epidemiologic surveys. From literature data, it appears that a higher intake of polyunsaturated fatty acids (linoleic acid) is related to an increase in mammary tumorigenesis in animal studies in which chemical carcinogens like dimethylbenz[a]anthracene are used as tumor initiator. Mostly the latency period of these chemically induced models in rather short. In this study, the Bald/c-MMTV (mouse mammary tumor virus) mouse strain was chosen as an animal model: MMTV leads to tumor initiation, and dietary factors influence tumor promotion over a relatively long latency period. The mice were fed diets with two fat concentrations: a high [36% of energy (en%)] or low (16 en%) fat level; fat was isocalorically replaced by carbohydrates (cornstarch). At both dietary fat levels, linoleic acid was given at four levels: 2, 3, 6, and 10 en%. Linoleic acid-rich fat was isocalorically replaced by oleic acid-rich fat. The diets were consumed ad libitum over a lifetime. Animals were euthanized as soon as mammary tumor diameter was > or = 1 cm or when the animals were in a poor clinical condition. The incidence of mammary tumors at 18 months was significantly higher in one group only: 36 en% fat and 2 en% linoleic acid. This group also showed the shortest mean latency period for mammary tumor development. Mean mammary tumor incidence was higher and mean onset time shorter in the four high-fat groups than in the low-fat groups. No (linear) dose-response relationship between dietary linoleic acid concentration and mammary tumor incidence and latency period was observed. This indicates that a higher dietary linoleic acid intake does not increase the incidence or shorten the latency period of breast cancer in the Balb/c-MMTV mouse strain at two different dietary fat levels. PMID- 8710686 TI - Serum carotene, vitamin A, and vitamin C levels in breast cancer and cancer of the uterine cervix. AB - Levels of carotene, vitamin A, and vitamin C measured in the serum of patients with cancer of the breast and uterine cervix were compared with levels in healthy controls and patients with benign diseases of the breast and cervix. Serum ascorbate levels were significantly lower in patients with benign diseases of the breast and cervix than in controls. In cancer patients, there was a significant trend of lower serum vitamin levels with increasing stage of the disease. PMID- 8710687 TI - Dietary sucrose, glucose, fructose, and starches affect colonic functions in rats. AB - To study the effect of dietary sugars and starches on parameters linked to colon carcinogenesis, female Sprague-Dawley rats were fed for one month five different diets containing sucrose, glucose, fructose, cornstarch, or Hylon 7, a starch with a high amylose content. After this period, colon proliferation, assessed by [3H] thymidine incorporation in vitro, was higher (p < 0.05) in rats fed sucrose than in rats fed glucose, fructose, or cornstarch [labeling index was 7.17 +/- 0.75, 5.03 +/- 0.07, 4.55 +/- 0.72, 4.00 +/- 0.70, and 5.89 +/- 1.05 (SE) in sucrose, glucose, fructose, cornstarch, and Hylon 7 diets, respectively]. Cecal pH was lower in rats fed cornstarch and Hylon 7 than in rats fed sucrose, glucose, or fructose. Content of short-chain fatty acids (SCFAs) in the cecum was higher in rats fed Hylon 7 than in those fed glucose and fructose. In conclusion, glucose and fructose, compared with sucrose, decrease mucosal proliferation and may be considered protective factors in colon carcinogenesis, although they do not affect SCFA production and cecal pH. On the contrary, Hylon 7 does not change mucosal proliferation but increases SCFAs and lowers cecal pH, two conditions associated with a lower risk of colon cancer. PMID- 8710688 TI - Effects of repeated boluses of sucrose on proliferation and on AOM-induced aberrant crypt foci in rat colon. AB - Colonic mucosal proliferation, aberrant crypt foci (ACF) induction, and fecal bile acids, parameters connected to the risk of colon cancer development, were studied in female F344 rats treated with starch or sucrose boluses or with a sucrose diet. Cell proliferation was higher in animals treated with a single sucrose bolus than in those given a starch bolus (15 g/kg body wt), with 4.3 +/- 0.64 and 2.17 +/- 0.57 (SE) mitotic figures (MF) per crypt in the sucrose and starch bolus groups, respectively (p < 0.01). When azoxymethane (AOM, 20 mg/kg) was administered 24 hours after a single sucrose or starch bolus, the number of ACF per colon after 30 days was higher in the sucrose bolus group [107.5 +/- 9.5 (SE)] than in the starch bolus group (67.8 +/- 0.9, p < 0.01). In additional experiments, colon cell proliferation (MF/crypt) was higher in rats given boluses of sucrose three times per week for 40 days after AOM (20 mg/kg) [5.9 +/- 0.7 (SE)] than in rats given starch boluses (2.96 +/- 0.4) or fed sucrose continuously (3.6 +/- 0.5). On the contrary, after 40 days of dietary treatment, the number, dimension, and percentage of ACF secreting sulfomucins and sialomucins were not varied among these three groups. However, the percentage of "large ACF" (ACF with > or = 4 crypts) secreting sialomucins or predominantly sialomucins was higher (p < 0.05) in the sucrose bolus group than in the starch group. The concentration of fecal bile acids and long-chain fatty acids was the same in the sucrose and starch groups, but the concentrations of deoxycholic and oleic acid were significantly higher in the sucrose bolus group. In conclusion, the administration of sucrose as a bolus had a stronger effect than continuous sucrose feeding on some parameters related to colon carcinogenesis and might be considered a risk factor in colon carcinogenesis. PMID- 8710689 TI - The effect of Lactobacillus GG on the initiation and promotion of DMH-induced intestinal tumors in the rat. AB - Male Fischer 344 rats were fed a 20% or a 5% corn oil diet and were injected subcutaneously with dimethylhydrazine (DMH) weekly for 16 weeks. In addition, an approximately equal number of animals challenged with DMH were fed daily, until the end of the study, 2 x 10(10) Lactobacillus casei subsp. rhamnosus strain GG starting three weeks before DMH administration or after the ninth weekly injection. The feeding of the Lactobacillus GG before and during carcinogen treatment resulted in a significant decrease in the incidence of colon tumors and the number of small intestinal and colon tumors per tumor-bearing animal for rats fed a 20% corn oil diet. This decrease in tumor incidence or number of tumors was not seen when animals were fed the Lactobacillus after the ninth week of carcinogen treatment. Animals fed a 5% corn oil diet had a lower tumor incidence and number of tumors resulting from the decrease in dietary fat; in addition the feeding of Lactobacillus GG before the carcinogen challenge resulted in a lower incidence of colon tumors. These studies show that a specific strain of L. casei subsp. rhamnosus designated GG can interfere with the initiation or early promotional stages of DMH-induced intestinal tumorigenesis, and this effect is most pronounced for animals fed a high-fat diet. PMID- 8710691 TI - Women and social development. PMID- 8710690 TI - Modulatory influence of alcoholic extract of Ocimum leaves on carcinogen metabolizing enzyme activities and reduced glutathione levels in mouse. AB - The present study reports the modulatory influence of alcoholic extract from the leaves of Ocimum sanctum on the activities of cytochrome p-450, cytochrome b5, and aryl hydrocarbon hydroxylase enzymes in the liver and glutathione-S transferase and reduced glutathione level in the liver, lung, and stomach of the mouse. Oral treatment with the leaf extract at 400 and 800 mg/kg body wt for 15 days would significantly elevate the activities of cytochrome p-450 (p < 0.05), cytochrome b5 (p < 0.01, p < 0.001), aryl hydrocarbon hydroxylase (p < 0.05), and glutathione S-transferase (p < 0.05, p < 0.01), all of which are important in the detoxification of carcinogens as well as mutagens. Moreover treatment with 400 and 800 mg/kg body wt of Ocimum extract for 15 days also significantly elevated extrahepatic glutathione-S-transferase (p < 0.05, p < 0.01). The reduced glutathione level was also elevated by treatment with the leaf extract in liver, lung, and stomach tissues (p < 0.01, p < 0.001). Mice fed a diet containing 0.75% butylated hydroxyanisole (positive control) revealed no alteration in the basal hepatic cytochrome p-450 and aryl hydrocarbon hydroxylase level, but hepatic cytochrome b5 and glutathione S-transferase activity in hepatic and extrahepatic organs were elevated in a time-responsive manner (p < 0.05, p < 0.001). The observations suggest further exploitation of the Ocimum leaf extract or its active principle(s) for the chemoprevention of chemical carcinogenesis in different animal model systems. PMID- 8710692 TI - World health day: in point of fact. A follow up feature. PMID- 8710693 TI - The lady with the lamp: learning from the pioneer of nursing. A Nurses' Day tribute. PMID- 8710694 TI - The panacea of comprehensive practice groups. PMID- 8710695 TI - An in vitro shear bond strength study of enamel/dentin bonding systems on enamel. AB - The purpose of this study was to compare the enamel shear bond strengths achieved with four acid conditioners employed by current enamel/dentin bonding systems (maleic acid, citric acid, nitric acid, oxalic acid) with a 37% phosphoric acid etching technique. The study also compared enamel shear bond strengths between the manufacturers' enamel/dentin conditioner used with an unfilled enamel bonding resin. The facial surfaces of 135 bovine incisors were ground flat and divided into nine test groups of n=15. Conditioning and bonding procedures were carried out following manufacturers' instructions. The phosphoric acid groups were etched for 15 seconds, rinsed for 30 seconds, and dried for 20 seconds with compressed air. All bonding was accomplished at a constant temperature of 21 degrees C and a relative humidity of 60%. A single composite restorative resin (Z-100) was used with all specimens to eliminate variables between composite materials. All specimens were thermocycled 2500 times (5 to 45 degrees C) and had a total storage time of 21 days prior to shear testing on an Instron at a crosshead speed of 1.0 mm/minute. Shear strengths were calculated by dividing load at failure by specimen area. A light microscope was used to determine failure mode. The data were subjected to Bartlett's test for homogeneity of variance and were found not to be homogeneous. The Welch Test was applied and indicated that the treatments used had influence on bond strength at P<0.05. There was no significant difference in bond strength values between traditional phosphoric acid/enamel bonding resin and Mirage Bond Dentin and Enamel Adhesive, Clearfil Liner Bond System, and Scotchbond Multi-Purpose Dental Adhesive System. PMID- 8710696 TI - The effect of amalgam surface preparation on the shear bond strength between composite and amalgam. AB - The aim of this study was to investigate the effect of amalgam surface preparation on the short-and long-term in vitro shear bond strength between a dental amalgam (Valiant) and two dental composites (Herculite, Z100) mediated by three universal bonding agents (All-Bond 2, Amalgambond, and Scotchbond Multi Purpose). Cylinders of dental composite resin were formed on, and bonded to flat amalgam surfaces. Prior to bonding, the amalgam surfaces were: a) ground flat, with the grinding lines oriented parallel to the direction of the shear stress; b) ground flat, with the grinding lines oriented perpendicular to the direction of the shear stress; or c) ground flat and then sandblasted using 50 micrometer Al2O3. A computerized Universal Testing Machine was used to determine the 1-, 7-, and 30-day shear bond strength of 162 samples (six per group) that were stored in distilled water at 37 degrees C. The results, which were statistically analyzed by performing a three-way analysis of variance (ANOVA) and 27 one-way ANOVA followed by modified (Bonferroni) t-tests for between-group comparisons (alpha = 0.05), have shown that: 1) sandblasting resulted in higher shear bond strength than grinding, except for 1-day All-Bond 2, where no differences were identified; 2) there was no difference in the shear bond strength between samples with parallel or perpendicular ground surfaces for a given bonding system and storage time; 3) for All-Bond 2, the 7-day shear bond strength of parallel and perpendicular ground samples dropped to half of the 1-day values, while the sandblasted samples showed a constant, high shear bond strength over 30 days, 4) for Amalgambond, the shear bond strength of parallel, perpendicular, and sand blasted samples was constant over 30 days; and 5) for Scotchbond Multi-Purpose, a sharp drop in the 7-day shear bond strength was recorded for all the samples, irrespective of amalgam surface preparation. PMID- 8710697 TI - Bond strength of composite to air-abraded enamel and dentin. AB - Human enamel and dentin were prepared with an air abrasion unit (KCP-2000), using two particle sizes (27 micrometers and 50 micrometers) of aluminum oxide. In vitro tensile bond strengths of a composite resin were determined after three enamel and three dentin surface treatments. Enamel treatments were: air abraded only, E-1; air abraded + adhesive, E-2; air abraded + acid etch + adhesive, E-3. Dentin surface treatments were air abraded only, D-1; air abraded + adhesive/no primer, D-2; air abraded + primer + adhesive, D-3. Etched enamel and dentin prepared with 600-grit SiC paper and adhesive served as controls. There were 10 replications for each condition. A dentin bonding system (Optibond) and a composite resin (Herculite XRV) were bonded to treated surfaces by light curing in an inverted, truncated cone die with a bond diameter of 3 mm. Samples were stored at 37 degrees C and 100% relative humidity for 24 hours and debonded in tension using a Universal Testing Machine at a 0.05 cm/min crosshead speed. Based on analysis of variance, there was no statistical difference between 27 micrometers and 50 micrometers aluminum oxide abrasive for both enamel and dentin. For enamel bond strengths, E-2 was significantly higher than E-1, and E-3 was significantly higher than E-1 and E-2. E-1 and E-2 were significantly lower than the control, while E-3 was not significantly different from the control (P < or = 0.05). For dentin bond strengths, D-2 was significantly higher than D-1, and D-3 was significantly higher than D-1 and D-2. All treatments except D-3 were significantly lower than the control (P < or = 0.05). Air-abrasion treatment of enamel and dentin alone resulted in reduced in vitro bond strengths as compared to etched enamel and dentin prepared with dentin adhesive and dentin primer. PMID- 8710698 TI - Repairability of polyacid-modified composite resin. AB - The purpose of this study was to evaluate the effect of time and surface treatment on the resulting shear bond strength of a repaired polyacid-modified composite resin restorative material (VariGlass VLC). Seventy-two specimens of VariGlass were prepared in cavities (2 mm X 7 mm) cut into acrylic resin cylinders. The specimens were then divided into three groups of 24. For each of the three groups, 12 of the specimens were repaired 5 minutes after the cavities were initially filled, while 12 specimens were repaired 1 week after the initial fill. The VariGlass surfaces were treated in one of the following ways: no treatment; primer (30 seconds) and adhesive (light activated for 10 seconds); or 37% phosphoric acid etchant (30 seconds), primer (30 seconds), and adhesive (light activated for 10 seconds). Repairs were made using a split polytetrafluoroethylene mold (3 mm X 5 mm) mounted over the exposed VariGlass surface. After bonding a cylinder of VariGlass to the treated surfaces, the specimens were thermocycled 500 times (5 degrees C and 55 degrees C water baths), stored in distilled water for 1 week, and loaded to failure in shear at 0.5 mm/min. Data were analyzed using two-way ANOVA and a Student-Newman-Keuls multiple comparison test. Results indicated that the bond strength of specimens repaired at 5 minutes was significantly higher when no surface treatment was used prior to repair. Bond strength of specimens repaired at 1 week was unaffected by surface treatment. Time of repair significantly affected the no-treatment and the primer-adhesive groups. PMID- 8710699 TI - Microleakage of gold casting repairs with different materials as quantified by a helium gas system. AB - Inadequate adaptation of a filling material to a gold crown can promote the passage of bacteria; thus, recontamination of sound dentin and/or the pulp canal space is feasible. The aim of this study was to determine the marginal microleakage between two different amalgams (Tytin and Valiant PHD-XT), three different composites (Tetric, Charisma, and Polofil Molar), and one glass-ionomer cement (Ketac Silver) and gold cast crowns using a helium gas microleakage method. In order to standardize the research parameters, gold washers with standardized dimensions were used as study models together with a helium leakage testing device. Standardized cavities were filled according to the manufacturers' recommendations with the different materials. The amount of helium passing the marginal interface between the fillings and cavities was measured with a mass spectrometer 48 hours after the fillings were placed and 100, 1000, and 2000 thermocycles (5 degrees C-55 degrees C). The results showed that amalgam allowed the least microleakage. Ketac Silver showed the greatest microleakage. Statistically significant differences were found between the composites and both amalgams and Ketac Silver between the 48-hour and 100-thermocycling groups. Yet, Ketac Silver showed a significant ascending tendency when compared to the composites and amalgams after 100, 1000, and 2000 thermocycles. PMID- 8710700 TI - In vivo adhesive interface between resin and dentin. AB - V-shaped cervical cavities prepared in monkey teeth were restored with several dentin bonding systems, and the in vivo resin-dentin interfacial structures were observed under the scanning electron microscope using an argon-ion etching technique. The hybrid layer could be clearly observed; its depth was dependent on the conditioner/primer used and tended to be thinner at the deep part of the cavity. Resin tags were also clearly observed, and their structure at the inner part was noted to be rougher than that closer to the tubule orifices. When 37% phosphoric acid gel was used, the tags in the deeper parts of the cavity were much rougher. Hemispherical and spherical roughened structures were observed directly above the tubule orifices, which were thought to be a mixture of dentinal fluid and bonding resin that had flowed out from the tubules. The acidic primer containing maleic acid and HEMA could not remove the smear plugs, and undissolved smear particles were observed in the tubules. PMID- 8710701 TI - Operative dentistry education: the pendulum ... and the pit. PMID- 8710702 TI - The postamalgam age. PMID- 8710703 TI - A histopathological study of direct pulp capping with adhesive resins. AB - This study evaluated the histological pulp responses of four different adhesive resin systems placed directly on exposed pulp tissue. Gap formation between the respective resin and cavity walls was observed and correlated to the pulpal response. Occlusal cavity preparations creating mechanical pulp exposures were prepared in molars of wistar rats. Preparations were filled using one of the following resin systems: 1) Superbond C&B system, 2) Clearfil Liner Bond system, 3) Tokuso Light Bond system, and 4) Scotchbond Multi-Purpose One All system. Although there were no significant differences in the pulpal responses after 3 days, significant differences were observed after 7 days. Results showed that the Superbond C&B system induced only a slight initial reaction, while secondary dentin was observed at the 30-day postoperative evaluation period. The Clearfil Liner Bond system showed a slight initial reaction to the materials, but exhibited slight pulpal necrosis and formation of secondary dentin at the 90-day evaluation period. The Tokuso Light Bond and One All systems showed severe pulpal reactions at all evaluation periods with no formation of secondary dentin. Microleakage formation correlated with the histopathological responses. Superbond C&B exhibited no microleakage, and only slight microleakage was observed when the Clearfil Bond system was used. The Tokuso and One All systems exhibited a wide area of microleakage. Further in vivo study evaluating the clinical use of adhesive resins for pulp capping is necessary to determine which resin systems may be used for direct pulp capping without incurring severe damage to pulpal tissue. PMID- 8710704 TI - Microleakage of new dentin bonding systems using human and bovine teeth. AB - The objective of this study was twofold: to evaluate the microleakage behavior of three dentin bonding systems and to determine if bovine teeth are comparable substrates to human teeth when studying the microleakage of various materials. The materials evaluated were Scotchbond Multi-Purpose Adhesive, Prisma Universal Bond 3, and All-Bond 2. All three bonding systems were used in combination with Prisma APH hybrid composite for comparison of microleakage behavior. Sixty class 5 preparations were cut at the cementoenamel junction for groups containing 30 human and 30 bovine teeth. A 1 mm 45 degree bevel was placed at the enamel margin. Teeth were grouped according to the dentin bonding system used and then restored according to the manufacturer's directions. After restoration, the teeth from each group were stored in distilled water at 37 degrees C for 3 days. The teeth were then thermocycled between 4 degrees C and 58 degrees C for 100 cycles and returned to distilled water at 37 degrees C for an additional 4 days. The teeth were then sealed with nail polish up to 1 mm from the margins of the restoration and placed in 45Ca isotope for 2 hours. The teeth were then sectioned and placed on x-ray film to produce autoradiographs. Microleakage was evaluated for the enamel and dentin margins separately using the following scale: 0 = no leakage, 1 = penetration of isotope to less than 1/2 the distance to the axial wall, 2 = penetration of isotope greater than 1/2 of the distance to the axial wall but short of the axial wall, and 3 = penetration of isotope to the axial wall or beyond. The materials were compared to each other using the Mann-Whitney U and Kruskal-Wallis tests. The gingival margins were compared to the incisal margins for all materials. No statistically significant differences in microleakage were revealed between the incisal and gingival location for human substrates, but there was statistically significant greater gingival microleakage for bovine substrates. All-Bond 2 leaked significantly more than Scotchbond Multi Purpose for human substrates at the incisal margin. All-Bond 2 had significantly more microleakage than Prisma Universal Bond 3 at both dentin and enamel margins for the bovine substrate. There were no statistically significant differences in microleakage among the bonding systems for the human substrate. No statistically significant differences between the microleakage behavior of human and bovine substrates were found. These results support the use of bovine teeth for in vitro microleakage studies. PMID- 8710705 TI - Shear bond strength of resin-mediated amalgam-dentin attachment after cyclic loading. AB - Several in vitro studies have suggested that a resin-mediated attachment mechanism can provide tooth reinforcement with silver amalgam restorations. This study investigated the ability of this attachment to withstand simulated occlusal function. Forty bovine incisors were divided into four groups. Prepared tooth surfaces were treated with a dentin bonding system (All-Bond 2) followed by amalgam (Tytin) condensation into a split mold. Samples were tested for shear bond strength after no cyclic loading or after 1000 and 5000 cycles with a weight of 700 grams (1.5 lb) at one strike per second. Mode of failure was assessed by microscopic investigation of fracture sites. The 24-hour shear bond strength (7.11 +/- 1.93 MPa) of Tytin spherical amalgam to dentin was progressively reduced after cyclic loading. Bond failure invariable occurred at the interface between the adhesive resin and amalgam. It was concluded that occlusal function may significantly reduce tooth reinforcement with bonded amalgam restorations. PMID- 8710706 TI - Mechanical properties and clinical performance of a gallium restorative material. AB - In a combination clinical/laboratory evaluation, a gallium restorative alloy (Galloy) was assessed for six mechanical properties: compressive strength, diametral tensile strength, creep, hardness, wear, and dimensional change. Laboratory tests indicated that the gallium alloy was similar to either Tytin or Dispersalloy in most respects, with the exception of lower wear and higher 7-day diametral tensile strength. In the clinical trial, all Galloy restorations were lined with a resin to prevent moisture contacting the material during the first hours after placement. All gallium restorations--class 1's only--were intact, and no teeth were fractured at the end of 1 year. There was little difference found at the 1-year evaluation compared to the 6-month results. The surface of the gallium alloy was generally rough, and many restorations exhibited some tarnish on the surface. It should be pointed out that this product has not been FDA approved at the present time and is not on the market. PMID- 8710707 TI - Evaluation of factors affecting the accuracy of impressions using quantitative surface analysis. AB - Impression material goes from a plastic to an elastic state during setting. Movement of the impression and excessive seating pressure during this transition can cause distortion in the impressions. The purpose of this study is to determine if the impression distortion is related to movement during setting or to distortion of the putty phase in the two-step impressioning technique. A master model of a maxillary quadrant of teeth was impressed using four different procedures: 1) one-step technique without movement (1S-NM); 2) one-step technique with movement (1S-M); 3) two-step technique without movement (2S-NM); and 4) two step technique with movement (2S-M). An artificial oral environment and surface analysis technique of the Minnesota Dental Research Center for Biomaterials and Biomechanics were used to produce the impressions and measure their accuracy. A digitized image of the first premolar of the master model was aligned with a digitized image of the first premolar of each epoxy model using AnSur. The root mean squared difference (RMS) between the aligned images is a measure of the distortion. The corresponding RMS values for the different methods were: 1S-NM = 23.7 +/- 9.21; 1S-M = 20.4 +/- 3.9; 2S-NM = 20.5 +/- 7.7; 2S-M = 21.3 +/- 4.4. Statistical analysis using a two-way analysis of variance showed no difference at the 0.05 level of significance. Pairwise comparison using the Tukey method showed that neither technique (one-step vs two-step) nor movement is a significant factor. These results showed that low seating pressure will not cause any greater distortions in the two-step impression technique than in the one-step technique, and minor movement during the setting of the impression material will no cause distortion. PMID- 8710708 TI - It could happen. PMID- 8710709 TI - Revised pediatric HIV classification system. PMID- 8710710 TI - Pediatric HIV infection and its oral manifestations: a review. PMID- 8710711 TI - Oral findings in asymptomatic (P-1) and symptomatic (P-2) HIV infected children. PMID- 8710712 TI - Oral soft tissue manifestations and CD4 lymphocyte counts in HIV-infected children. AB - This study investigated the prevalence of oral soft tissue lesions in children infected with HIV and the relationship of CD4 lymphocyte levels with the prevalence of those lesions. Sixty HIV-positive children enrolled in the Children's Hospital AIDS Program (age 5.8 +/- 3 years) were selected for study. Only five subjects (8%) had healthy gingiva and a low mean plaque index (22%). The remainder had gingivitis or periodontitis with relatively high plaque indices (47, 55, and 94%, respectively). A declining CD4 lymphocyte count (1357 to 35) was associated with an increasing severity of gingival disease. PMID- 8710714 TI - Cancer in the HIV-infected child. PMID- 8710713 TI - Risk factors for HIV-related orofacial soft-tissue manifestations in children. AB - A retrospective review of the medical records of 492 children perinatally exposed to the human immunodeficiency virus (HIV) compared the prevalence of orofacial soft-tissue manifestations in HIV-infected and noninfected children, identified risk factors for occurrence of orofacial lesions in HIV-infected children, and investigated specific orofacial lesions as indicators of progression of HIV disease. Application of eligibility criteria and the Centers for Disease Control classification of pediatric HIV infection resulted in selection of a study group of 91 HIV-positive children and a control group of 185 HIV-seronegative children who had seroreverted. Analysis of oral lesions showed that 67% of the study group and 8% of the control group had oropharyngeal candidiasis (OPC), 4% of the study group and 0% of the control group had parotid enlargement, and 3% of the study group and 0% of the control group had herpes simplex; all three differences were significant at P < 0.04. No statistically significant association was found between OPC and the risk factors of gender, ethnicity, or mode of delivery (vaginal versus cesarean). However, OPC was associated significantly with all progression markers examined: failure to thrive, use of antiretroviral agents, lower CD4 counts, and development of acquired immunodeficiency syndrome (AIDS). Orofacial manifestations are common in pediatric HIV infection and may serve as markers of infection and predictors of progression of HIV disease to AIDS. PMID- 8710715 TI - Caries experience and cariogenic markers in HIV-positive children and their siblings. AB - The purpose of this cross-sectional, masked study was to compare the oral status of perinatally HIV-infected children with their uninfected siblings living in the same environment. A secondary purpose was to compare HIV-positive children for differences in oral health with respect to disease advancement. One hundred forty seven children were examined in their homes and meeting places, using NIH criteria for caries diagnosis. Significant differences were found in the number of caries-free children (P < 0.05), past caries experience (P < 0.003), subsurface demineralizations (P < 0.0001), and caries-related bacteria (P < 0.05). However, differences in caries prevalence were not found in the 3- to 6 year-old subgroup. Caries prevalence (P < 0.001) and levels of caries-related flora in saliva were correlated to years since diagnosis (mutans streptococci P < 0.008, lactobacilli P < 0.02). Children with a more advanced disease stage had significantly more caries (P < 0.02). Among the HIV-infected children, the frequency of carbohydrate intake was clearly correlated to caries (P < 0.003) and to lactobacilli levels (P < 0.0001). It is concluded that children with perinatally acquired HIV are at greater risk for caries than their siblings, more so with advancing disease. PMID- 8710716 TI - Compliance is poor among HIV-infected children with unmet dental needs. PMID- 8710717 TI - Evaluation of an alum-containing mouthrinse in children for plaque and gingivitis inhibition during 4 weeks of supervised use. AB - Aluminum salts have demonstrated activity against oral bacteria and also have shown indications of inhibiting plaque and gingivitis. The aims of this clinical trial were to determine the effects of daily supervised rinsing with a specially formulated, alum-containing mouthrinse on existing plaque and gingivitis in children and to monitor its effect on the oral tissues and its acceptability to subjects. Using a double-blind, parallel design, 48 sixth-graders rinsed once daily for 30 sec under supervision with either a placebo or a mouthrinse containing 0.02 M alum while continuing their normal oral hygiene habits. Plaque and gingivitis were assessed and intraoral examinations were performed at 0, 2 and 4 weeks. The alum mouthrinse significantly (P < 0.05) reduced the amount of plaque relative to the placebo after both 2 and 4 weeks. Gingivitis and plaque thickness also were decreased, but the differences did not attain significance. No evidence of deleterious effects to the oral tissues was observed and the alum mouthrinse was well accepted by the children. This trial demonstrated that daily use of an alum-containing mouthrinse was safe and produced a significant effect on plaque that supplemented the benefits of daily toothbrushing. Thus, topically applied aluminum may have potential applications in preventive dentistry for controlling plaque. PMID- 8710718 TI - Subluxation injuries of maxillary primary anterior teeth: epidemiology and prognosis of 207 traumatized teeth. AB - This study investigated the epidemiology, sequelae, and prognosis of subluxation injuries to the maxillary primary anterior dentition. Data were collected from dental records at the Montreal Children's Hospital, Montreal, Canada, of patients sustaining trauma between 1982 and 1993. The study group consisted of 207 teeth in 134 patients, 81 males and 53 females. The age of the patients ranged from 0.8 years to 7.5 years, with a mean of 3.5 years. The highest incidence of trauma was in males between the ages of 3 and 4 years, and in females between 1 and 3 years. The highest incidence of trauma (66.2%) involved the primary central incisors. The most common cause of trauma (52%) was simple falls indoors. Occlusal or periapical radiographs were the radiographic view of choice unless an anterior nasal spine fracture was suspected, in which case a lateral projection was exposed. Approximately 64% of the study teeth had a mobility ranging from 0.6 to 1.5 mm. Treatment varied from no treatment (80% of teeth) to extraction. Post traumatic evaluations were distributed into six common time intervals used by dentists for follow up. The results indicated that patient discomfort and occlusal interference were not common. Discoloration and pulpal calcification increased with time. Mobility decreased with time, with the majority of teeth returning to a normal physiologic range. External resorption may have been present, but was rare. Treatment or antibiotics were rarely needed at follow-up visits. Overall, these teeth responded positively, and there was a low morbidity associated with subluxation injuries. PMID- 8710719 TI - Access to dental care for Medicaid children in Connecticut. PMID- 8710720 TI - Segmental odontomaxillary dysplasia (hemimaxillofacial dysplasia): case report. PMID- 8710721 TI - Update on American Board examination results. PMID- 8710722 TI - Advances in the management of acute asthma in children. PMID- 8710723 TI - Child sexual abuse. AB - Multiple obstacles can hinder the medical evaluation of suspected child sexual abuse in pediatric primary care. The need for diagnostic accuracy is high. Knowledge of sexual abuse risk factors, an understanding of the victimization process, and awareness of the varied clinical presentations of sexual abuse can be of assistance. Open-ended questioning of the suspected victim is the most critical component of the evaluation. Skillful medical interviewing requires time, training, patience, and practice. Pediatricians lacking any of these four requirements should defer interviewing in sexual abuse cases to other professionals. Abnormal physical findings from sexual abuse are uncommon. Colposcopy has assisted pediatricians greatly in reaching consensus regarding diagnostic physical findings. Cases of acute sexual assault require familiarity with the forensic rape examination, STD screening and prophylaxis, and pregnancy prevention. Victimization from sexual abuse continues long after the abusive acts end, often requiring long-term therapeutic intervention. An emerging standard of care for medical evaluations of suspected child sexual abuse recognizes the requirement for patience and compassion while retaining objectivity. The pediatrician's primary concern must be for the child's physical and emotional well-being. PMID- 8710724 TI - Quality improvement: an ACQIP exercise on the management of otitis media. PMID- 8710725 TI - Apparent life-threatening events. PMID- 8710726 TI - The hidden costs of keyhole care. PMID- 8710727 TI - A request for more hypnotics. PMID- 8710728 TI - A guide to non-opiate illicit drugs. PMID- 8710729 TI - Arguments for day case surgery. PMID- 8710730 TI - Preparing for the CRQ part II. PMID- 8710731 TI - A rational approach to hepatitis C. PMID- 8710732 TI - Surgical choices in colitis care. PMID- 8710733 TI - When to investigate and treat IBS symptoms. PMID- 8710734 TI - Diarrhoea in the returned traveller. PMID- 8710735 TI - Coeliac disease: looking out for easily reversible morbidity. PMID- 8710736 TI - Rectal bleeding: when is it right to refer? PMID- 8710737 TI - Microtubule-based motor proteins: new targets for enhancing drug delivery? PMID- 8710739 TI - Evidence for intestinal secretion as an additional clearance pathway of talinolol enantiomers: concentration- and dose-dependent absorption in vitro and in vivo. AB - PURPOSE: To evaluate carrier-mediated intestinal secretion of talinolol enantiomers in vivo and in vitro. METHODS: In clinical studies with i.v. and p.o. dosage of rac-talinolol (30 mg and 100 mg, resp.) performed in a small number of cholecystectomized patients total and partial clearances were determined on the basis of plasma, bile and urine concentrations. The dose-dependence of AUC was investigated in 12 healthy volunteers (25, 50, 100, and 400 mg rac-talinolol as single p.o. doses). Concentration-dependence of the permeability across Caco-2 cell monolayers included concentrations from 0.1 to 2.0 mM, inhibition by verapamil was tested at 0.5 mM. RESULTS: The total clearance as well as the apparent oral clearance (CL/F) were slightly higher for S-(-)- than for R-(+) talinolol. Calculation of the partial clearances showed that also the residual clearance was higher for the S- than for the R-enantiomer. In the healthy volunteers, CL/F increased with increasing doses, while the S/R ratio decreased approaching unity for the highest dose. Also the results from Caco-2 cell permeation studies yielded a clear concentration-dependence with decreasing stereoselectivity for the higher concentration range. Permeability of both enantiomers was considerably higher for b-->a than a-->b transport, however, this difference disappeared when verapamil was added. CONCLUSIONS: Although not very expressed, the detected stereoselectivities indicate a preferential absorption of R-(+)-talinolol in a lower concentration and dose range, which is most probably due to a moderate stereoselectivity at the carrier system involved in intestinal secretion. PMID- 8710738 TI - Recent advances in carrier-mediated hepatic uptake and biliary excretion of xenobiotics. AB - PURPOSE: Besides renal excretion, hepatic metabolism and biliary excretion are the major pathways involved in the removal of xenobiotics. Recently, for many endogenous and exogenous compounds (including drugs), it has been reported that carrier-mediated transport contributes to hepatic uptake and/or biliary excretion. In particular, primary active transport mechanisms have been shown to be responsible for the biliary excretion of anticancer drugs, endogenous bile acids and organic anions including glutathione and glucuronic acid conjugates. Primary active excretion into bile means the positive removal of xenobiotics from the body, and this elimination process is now designated as "Phase III" (T. Ishikawa, Trends Biochem. Sci., 17, 1992) in the detoxification mechanisms for xenobiotics in addition to Phase I by P-450 and Phase II by conjugation. METHODS: The transporters, which have been called P-glycoprotein (MDR), multidrug resistance related protein (MRP) and GS-X pump and which are believed to be involved in the primary active pumping of xenobiotics from the cells, are now known as the ATP-binding cassette (ABC) transporters. In this review, we first describe the HMG-CoA reductase inhibitor, pravastatin, as a typical case of a carrier-mediated active transport system that contributes to the liver-specific distribution in the body. RESULTS: Regarding biliary excretion, we have summarized recent results suggesting the possible contribution of the ABC transporters to the biliary excretion of xenobiotics. We also focus on the multiplicities in both hepatic uptake and biliary excretion mechanisms. Analyzing these multiplicities in transport is necessary not only from a biochemical point of view, but also for our understanding of the physiological adaptability of the living body in terms of the removal (detoxification) of xenobiotics. CONCLUSIONS: Clarification of these transport mechanism may provide important information for studying the pharmacokinetics of new therapeutic drugs and furthermore, leads to the development of the drug delivery systems. PMID- 8710740 TI - Characterization of the transport properties of a quinolone antibiotic, fleroxacin, in rat choroid plexus. AB - PURPOSE: It is reported that the cerebrospinal fluid (CSF) to plasma unbound concentration ratio of fleroxacin at steady-state is approximately 0.5 in experimental animals. These results can be accounted for by assuming the presence of an active transport system for the efflux of this compound across the choroid plexus. In the present study, the transport system for fleroxacin was characterized in isolated rat choroid plexus. METHODS: Choroid plexus was isolated from the lateral ventricles of rats. The accumulation of [14C] fleroxacin or [3H] benzylpenicillin by the choroid plexus was examined by the centrifugal filtration method. RESULTS: The accumulation of [14C] fleroxacin by the rat isolated choroid plexus was significantly inhibited by metabolic inhibitors (rotenone, 30 microM and carbonyl cyanide rho-trifluorometh oxyphenylhydrazone (FCCP), 100 microM) and sulfhydryl reagent (p chloromercuribenzenesulfonic acid (PCMBS), 100 microM). This accumulation was composed of a saturable component (Vmax = 240 pmol.min-1.microliter tissue-1, Km = 664 microM) and non-saturable one (P = 0.424 min-1.microliter tissue-1). Accumulation of fleroxacin was competitively inhibited by benzylpenicillin and probenecid with Ki values of 29 microM and 51 microM, respectively. These values are comparable with the Km of benzylpenicillin transport and the Ki of probenecid for the benzylpenicillin transport at the choroid plexus, respectively. Furthermore, fleroxacin inhibited competitively the accumulation of [3H] benzylpenicillin with a Ki of 384 microM, a value comparable with the Km of [14C] fleroxacin transport. CONCLUSIONS: Fleroxacin and benzylpenicillin showed mutual competitive inhibition, suggesting that both are transported via a common transport system in the choroid plexus and are pumped out from CSF into the circulation. PMID- 8710741 TI - The use of surfactants to enhance the permeability of peptides through Caco-2 cells by inhibition of an apically polarized efflux system. AB - PURPOSE: It has recently been reported that the permeability of peptides across Caco-2 cells, an in vitro model of the intestinal mucosa, was limited by an apically polarized efflux mechanism. Since surfactants (e.g. Cremophor EL, Polysorbate 80) have been reported to inhibit similar efflux systems in tumor cells, we determined whether they could enhance the permeability of peptides across monolayers of Caco-2 cells. METHODS: The transport studies of [3H] mannitol and [14C]-model peptides were carried out across the Caco-2 cell monolayers. TEER values were determined using Voltohmmeter with STX-2 electrode and the equilibrium dialysis studies were conducted using side-by-side dialysis apparatus with cellulose ester membranes. RESULTS: Initially, [3H]-mannitol flux studies were conducted to find concentrations of the surfactants that did not cause damage to the cell monolayer. Based on these studies, Polysorbate 80 and Cremophor EL were selected for further study. The fluxes of [14C]-AcfNH2 (a nonsubstrate for this efflux system) and [14C]-Acf(N-Mef)2NH2 (a substrate for this efflux system) were then measured in the absence and presence of the two surfactants. The permeability of [14C]-AcfNH2 was not affected by the surfactants, while that of [14C]-Acf(N-Mef)2NH2 increased with increasing concentrations of surfactants and then decreased. For example, the Pe values for [14C]-Acf(N-Mef)2NH2 were 3.75 x 10(-6), 8.58 x 10(-6), 10.29 x 10(-6), 7.48 x 10(-6), and 1.46 x 10(-6) cm/sec with Cremophor EL concentrations of 0, 0.01, 0.1, 1 and 10% w/v, respectively. This bimodal effect of surfactants on the Caco 2 cell permeability of this peptide was shown to be due to the interactions between the peptide and micelles at higher concentrations of surfactants, which were demonstrated by the equilibrium dialysis experiments. CONCLUSIONS: These results suggest that surfactants, which are commonly added to pharmaceutical formulations, may enhance the intestinal absorption of some drugs by inhibiting this apically polarized efflux system. PMID- 8710742 TI - Diffusivity of bile salt/phospholipid aggregates in mucin. AB - PURPOSE: The objective of this study is to evaluate the effect of the mucous layer on the transport of the drug-solubilizing bile salt/phosphatidylcholine (BS/PC) aggregates. METHODS: The self-diffusion coefficient of BS/PC aggregates in bovine submaxillary mucin (BSM) was measured by Fourier-transform pulsed-field gradient spin-echo (FT-PGSE) 1H NMR spectroscopy. RESULTS: In spite of the complexity of the mixture, the FT-PGSE technique allowed the unambiguous determination of the diffusivity of PC and 1H2HO (HDO, natural abundance in D2O). With a series of BS/PC total lipid concentrations ranging from 1 to 7 g/dl, a progressive decrease in the effective diffusivity of HDO was observed with an increase in the both the BSM and total lipid concentration. The effective diffusivity of PC decreased with increasing lipid concentrations in the presence of mucin, while in the controls it increased. After correcting the effective diffusivity of PC for the obstruction effect of mucin, the size of the BS/PC mixed micelle was assessed. It appears that PC associates with BSM resulting in a decrease in the available PC for micellization. This reduces the average size of the mixed micelle within the mucous layer. CONCLUSIONS: The aggregation state of BS/PC micelle is altered by the presence of mucin which would have a direct impact on the transport of dietary lipid and solubilized drug through the aqueous boundary layer of the intestinal tract. PMID- 8710743 TI - The relative effect of Azone and Transcutol on permeant diffusivity and solubility in human stratum corneum. AB - PURPOSE: The purpose of this work was to analyse the mechanism of the enhancement of percutaneous penetration demonstrated by the known enhancers Azone and Transcutol. METHODS: Enhancer induced changes in the diffusivity and solubility of a model permeant (4-cyanophenol) in human stratum corneum were monitored (in vitro) using Attenuated total reflectance Fourier transform infra-red (ATR-FTIR) spectroscopy and compared to the gross effects of the enhancers on flux as measured using simple Franz-type diffusion cells. RESULTS: It has been shown by both the well-established Franz diffusion cell technique and the use of ATR-FTIR spectroscopy that the enhancers studied both increase the flux of cyanophenol across human skin in-vitro by a factor of approximately two. Furthermore, it has been demonstrated by ATR-FTIR that these enhancers are likely to exert their effects by different mechanisms. It is probable that Azone reduces the diffusional resistance of the stratum corneum and that Transcutol increases the solubility of the penetrant in this barrier. CONCLUSIONS: There is increasing interest in the apparently synergistic nature in which certain enhancers appear to work. The exact nature of these multiplicative and/or additive effects is not known although there are numerous suggestions in the current literature. The application of ATR-FTIR spectroscopy to such enhancing systems will allow the mechanisms of the observed enhancements to be probed in greater depth. PMID- 8710744 TI - Response surface method: a novel strategy to optimize iontophoretic transdermal delivery of thyrotropin-releasing hormone. AB - PURPOSE: To maximize the iontophoretic transdermal delivery rate of thyrotropin releasing hormone (TRH) facilitated by periodically monophase-pulsed current across excised skin. METHODS: The pH of the buffer, the ionic strength in the solution, the frequency of the periodically monophase-pulsed current and the current on/off ratio were chosen as the key variables. A response surface method was applied to optimize the transdermal delivery rate of TRH under different operational conditions. RESULTS: The optimum operating conditions were achieved via experimentation based on the response surface method by systematically adjusting the pH of the buffer, the ionic strength in the solution, the current amplitude, frequency and the active temporal ratio of the pulsed current. The rate of permeation of TRH crossing the skin during iontophoresis varied from two to ten-fold, depending on operating conditions. CONCLUSIONS: Only a few steps, two in this work, were needed to reach the optimal. The response surface near the region of the maximal point was thoroughly described with a quadratic function. A maximal transdermal rate of permeation of TRH, 103.2 micrograms h-1 cm-2, was obtained when the donor solution was at pH = 7.0, ionic strength = 0.037, and with a periodically monophase-pulsed current iontophoresis with duty cycle = 75%. The effect of pulse frequency was not statistically significant. PMID- 8710745 TI - Iontophoresis of bases, nucleosides, and nucleotides. AB - PURPOSE: To investigate whether transdermal iontophoresis may be potentially useful for delivery of oligonucleotide drugs, the electrotransport of representative bases (uracil and adenine), nucleosides (uridine and adenosine) and nucleotides (AMP, ATP, GTP and imido-GTP) across mammalian skin in vitro has been considered. METHODS: While the passive permeability of all compounds investigated (from 1 mM solutions at pH 7.4) was very low, the application of constant current iontophoresis (0.55 mA/cm2) significantly enhanced the transport of both charged and uncharged species. RESULTS: The efficiency of delivery depended only weakly upon lipophilicity, varied quite linearly with concentration (for AMP and ATP), was inversely sensitive to molecular weight, and was strongly influenced by charge. Neutral solutes were delivered better from the anode than the cathode, as expected; post-iontophoresis, passive permeabilities were greater than those of the untreated controls, suggesting that iontophoretically-induced changes in barrier function cannot be completely repaired in in vitro model systems. The triphosphate nucleotides, ATP and GTP, were essentially completely metabolized (presumably to their corresponding mono-phosphates) during their iontophoretic delivery, while imido-GTP was apparently resistant to enzymatic attack; however, comparison of the transport data from AMP and ATP suggested that ATP metabolism occurred primarily after the rate-limiting step of iontophoresis. CONCLUSIONS: The results obtained are consistent with the general patterns of behavior previously observed in investigations of amino acid and peptide electrotransport. It remains to be seen whether extension of the research described here to larger oligonucleotide species is a feasible long-term objective. PMID- 8710746 TI - Transdermal delivery of fentanyl by electroporation. I. Influence of electrical factors. AB - PURPOSE: Electroporation, a method of reversibly permeabilizing lipid bilayers by the application of an electric pulse, has been shown to induce increased transdermal passage of molecules. The aim of the present report was to study in vitro with hairless rat skin the potential of electroporation for transdermal delivery of fentanyl. RESULTS: The application of electric pulses can strongly promote transdermal delivery of fentanyl compared to passive diffusion through untreated skin. We also point out that the choice of the waveform of the electric pulses is important: at the same applied energy, a few exponentially-decaying (ED) pulses increased fentanyl permeation more than a few square-wave pulses and to the same extent as the repeated application of higher voltage-shorter duration ED pulses. A factorial design showed that the voltage, duration, and number of ED pulses allowed control of the quantity of drug transported through the skin. CONCLUSIONS: Skin electroporation could be a good way to improve the transdermal diffusion of fentanyl. PMID- 8710747 TI - Theoretical considerations on two equations for estimating the extent of absorption after oral administration of drugs. AB - PURPOSE: The amount of drug absorbed into portal blood after oral dosing (Dp.o,g) has been estimated using Fick's principle (Q-method), i.e., Dp.o,g = Qh x (AUCp.o,g--AUCp.o,c), where Qh is the portal blood flow rate, and AUCp.o,g and AUCp.o,c are the areas under the concentration-time curves of portal vein and systemic blood after oral dosing, respectively. However, this method may underestimate Dp.o,g, when the drug is subject to systemic intestinal elimination. An alternate equation (CL-method; Dp.o,g = CLs x AUCp.o,g) is described using a simple pharmacokinetic model, to estimate Dp.o,g in the presence of systemic intestinal elimination, where CLs is systemic clearance. METHODS: The model is composed of central, intestine and liver compartments, assuming that drug is eliminated by intestinal and/or hepatic pathways only. A comparison of both methods for estimating Dp.o,g was made using computer simulation or experimental data of phenacetin from the literature. RESULTS: The simulation study demonstrated that the Q-method underestimated Dp.o,g in the presence of significant intrinsic intestinal clearance, compared to the CL method. The similar results were observed using the experimental data of phenacetin. CONCLUSIONS: The CL-method can provide a better estimate of Dp.o,g, while the Q-method may underestimate Dp.o,g, when there is significant systemic intestinal elimination of drugs after oral administration. In addition, useful information for understanding the relationship between the extent of absorption and the first-pass effect by intestine and/or liver after oral dosing of drugs can be obtained from the present approach. PMID- 8710748 TI - Analysis of intestinal perfusion data for highly permeable drugs using a numerical aqueous resistance--nonlinear regression method. AB - PURPOSE: To develop, validate and apply a method for analyzing the intestinal perfusion data of highly permeable compounds using the Numerical Aqueous Resistance (NAR) theory and nonlinear regression (NAR-NLR) and to compare the results with the well-established Modified Boundary Layer (MBL) Analysis. METHODS: The NAR-NLR method was validated and the results were compared to the MBL analysis results using previously reported cephradine jejunal perfusion data. Using the Single Pass Intestinal Perfusion (SPIP) method, the concentration dependence of intestinal permeability was investigated for formycin B, proline, and thymidine, three compounds reported to be absorbed by carrier-mediated transport processes. The MBL and NAR-NLR analyses were then applied to the three sets of SPIP data. RESULTS: The results demonstrate that the intrinsic MBL transport parameters were highly variable and, in one case, the analyses failed to give a statistically significant Michaelis constant. The MBL mean dimensionless wall permeabilities (P*w) were greater than the NAR-NLR P*w and were also highly variable. In all cases, the NAR-NLR variability was significantly lower than the MBL variability. The extreme variability in the MBL calculated P*w is due to the sensitivity of P*w when the fraction of unabsorbed drug (Cm/Co) is low or, alternatively, when P*w approached the aqueous permeability, P*aq. CONCLUSIONS: The NAR-NLR method facilitates the analysis of intestinal perfusion data for highly permeable compounds such as those absorbed by carrier-mediated processes at concentrations below their Km. The method also allows for the use of a wider range of flow conditions than the MBL analysis resulting in more reliable and less variable estimates of intestinal transport parameters as well as intestinal wall permeabilities. PMID- 8710749 TI - Effects of transfection with the Cu, Zn-superoxide dismutase gene on xanthine/xanthine oxidase-induced cytotoxicity in fibroblasts from rat skin. AB - PURPOSE: The effects of transfection with the human Cu, Zn-superoxide dismutase (hSOD)4 gene on active oxygen-induced cytotoxicity in rat skin fibroblasts (FR) were studied for the purpose of developing the novel delivery system of hSOD using hSOD gene. METHODS: An expression plasmid for hSOD, pRc/RSV-SOD, was constructed and used to transfect FR cells. Xanthine (X)/xanthine oxidase (XO) system were used to generate active oxygen species. The effects of transfection with the hSOD gene on active oxygen-induced cytotoxicity were assessed by comparing the number of surviving cells and the level of lipid peroxidation in host and transformants after exposure to X/XO system. RESULTS: The cellular SOD activity in RSV-SOD cells transfected with pRc/RSV-SOD was significantly increased in comparison with host or RSV cells transfected with the pRc/RSV plasmid containing no hSOD gene as a control. Furthermore, Western blot analysis using an anti-hSOD antibody indicated the production of hSOD in RSV-SOD cells. On the other hand, although the numbers of surviving cells in both host and RSV-SOD cultures after exposure to X/XO system decreased in a time-dependent manner, the decrease in number of surviving RSV-SOD cells was less than that in host cells. In the presence of catalase, the decreases in number of surviving cells in both host and RSV-SOD cultures after exposure to the X/XO system were also less than those in the absence of catalase. However, the decreases in cell survival in RSV SOD cultures were significantly less than those in host cells in the presence of catalase. Furthermore, the levels of lipid peroxidation in RSV-SOD cells exposed to the X/XO system in the presence or absence of catalase were lower than those in host cells. These results indicated that the increase in cellular SOD activity by transfection with the hSOD gene protects cells from oxidative stress. CONCLUSIONS: Human SOD gene therapy may be useful for treatment of diseases in which oxidative tissue damage is produced. PMID- 8710750 TI - The effect of formulation on the antimicrobial activity of cetylpyridinium chloride in candy based lozenges. AB - PURPOSE: The purpose of this investigation was to determine the influence on the antimicrobial activity of cetylpyridinium chloride of the various components of the formulation of each of six candy based lozenges. METHODS: In vivo activity was investigated using six volunteers by determining the reduction in colony forming units recoverable from the oropharynx after sucking each lozenge separately on different days. In vitro determinations investigated the relative activity of aqueous solutions of the lozenges, the effect on activity of additional active ingredients, pH and lozenge base ingredients against separate inocula of each of the test organisms Staphylococcus aureus, Streptococcus pyogenes and Candida albicans. RESULTS: Both in vivo and in vitro results showed that the pH of the dissolved lozenge solution was the single most influential readily adjustable formulation parameter which significantly influenced the activity of cetylpyridinium chloride activity in candy based lozenges. CONCLUSIONS: Lozenges containing cetylpyridinium chloride as the active ingredient should be formulated at a pH greater than 5.5. PMID- 8710751 TI - Preparation and characterization of freeze-dried chitosan-poly(ethylene oxide) hydrogels for site-specific antibiotic delivery in the stomach. AB - PURPOSE: The purpose of this study was to develop novel drug delivery systems with pH-sensitive swelling and drug release properties for localized antibiotic delivery in the stomach. METHODS: The drug delivery systems were synthesized by crosslinking chitosan and poly(ethylene oxide) (PEO) in a blend to form semi interpenetrating polymer network (semi-IPN). Scanning electron microscopy was used to compare the surface and bulk morphology of the freeze-dried and air-dried chitosan-PEO semi-IPN. The hydrogels were allowed to swell and release the antibiotics--amoxicillin and metronidazole--in enzyme-free simulated gastric fluid (SGF, pH 1.2) and simulated intestinal fluid (SIF, pH 7.2) at 37 degrees C. RESULTS: Freeze-dried chitosan-PEO semi-IPN with a porous matrix had swollen extensively as compared to the air-dried hydrogel. The swelling ratio of freeze dried and air-dried chitosan-PEO semi-IPN after 1 h in SGF was 16.1 and 2.30, respectively. More than 65% of the entrapped amoxicillin and 59% of metronidazole were released from the freeze-dried chitosan-PEO semi-IPN after 2 h in SGF. CONCLUSIONS: The results of this study suggest that freeze-dried chitosan-PEO semi-IPN could be useful for localized delivery of antibiotics in the acidic environment of the gastric fluid. PMID- 8710752 TI - Relationship between gelation rate of controlled-release acetaminophen tablets containing polyethylene oxide and colonic drug release in dogs. AB - PURPOSE: We hypothesized that sufficient gelation of orally administered hydrophilic matrix tablets before they reach the colon could, as a result of continuous erosion of the gelated matrix, prevent the decrease in colonic drug release which normally occurs here. The purpose of this study was to elucidate the effect of gelation of hydrophilic matrices containing polyethylene oxide on colonic drug release in dogs using controlled-release (CR) acetaminophen tablets. METHODS: Two types of CR tablets were prepared, a slow gelling tablet (SG) and a rapid gelling tablet (RG) containing an extra highly water soluble filler. In vitro and in vivo performance were examined. RESULTS: SG and RG showed similar drug release behavior in vitro. In oral administration to dogs, the two formulations showed similar gastrointestinal transit, reaching the colon within 2 4 h after oral dosing. Further, they showed similar maximum plasma levels (Cmax) and time to Cmax (Tmax). In contrast, however, the two tablets produced different plasma levels from 2 h post-dosing, with plasma levels of RG higher than those of SG and with smaller individual variation. Directly observed colonic drug release behavior of RG was similar to in vitro drug release, whereas that from SG was suppressed. CONCLUSIONS: Colonic drug release is closely related to the gelation of hydrophilic matrix, and rapid gelation provides continuous in vivo drug release. PMID- 8710753 TI - Disposition characteristics of plasmid DNA in the single-pass rat liver perfusion system. AB - PURPOSE: To define the hepatic uptake mechanism of a plasmid DNA, we quantitated the uptake of pCAT (plasmid DNA encoding chloramphenicol acetyltransferase reporter gene fused to simian virus 40 promoter), a model plasmid, after a single pass through the perfused rat liver using albumin- and erythrocyte-free Krebs Ringer bicarbonate buffer (pH 7.4). METHODS: [32P]pCAT was introduced momentarily into this system from the portal vein as a bolus input or constant infusion mode, and the outflow patterns and hepatic uptake were evaluated using statistical moment analysis. RESULTS: The venous outflow samples had electrophoretic bands similar to that of the standard pCAT, suggesting that the plasmid is fairly stable in the perfusate during liver perfusion. In bolus experiments, pCAT was largely taken up by the liver and the uptake was decreased with increase in injected dose. Statistical moment analysis against outflow patterns demonstrated that the apparent volume of distribution of pCAT was greater than that of human serum albumin, indicating a significant reversible interaction with the tissues. The results of collagenase perfusion experiments suggest that the hepatic accumulation of pCAT occurred preferentially in the nonparenchymal cells (NPC). The amount of total recovery in the liver decreased substantially by preceding administration of polyinosinic acid, dextran sulfate, succinylated bovine serum albumin, but not by polycytidylic acid. This suggests that pCAT is taken up by the liver via scavenger receptors for polyanions on the NPC. In constant infusion experiments, the presence of 2,4-dinitrophenol and NH4Cl caused a significant increase in the outflow concentration of [32P]pCAT and decrease by half in the total hepatic recovery than that of plasmid DNA administered alone, suggesting that plasmid DNA may undergo internalization by the NPC. CONCLUSIONS: The liver plays an important role in the elimination of plasmid DNA and a successful delivery system will be required to avoid its recognition by the scavenger receptors on the liver NPC. PMID- 8710754 TI - Immunoliposomes as enzyme-carriers (immuno-enzymosomes) for antibody-directed enzyme prodrug therapy (ADEPT): optimization of prodrug activating capacity. AB - PURPOSE: Immuno-enzymosomes are tumor-specific immunoliposomes bearing enzymes on their surface. These enzymes are capable of converting relatively nontoxic prodrugs into active cytostatic agents. The enzyme beta-glucuronidase (GUS)4 was coupled to the external surface of immunoliposomes directed against ovarian carcinoma cells. This study aimed at optimization of the prodrug-activating capacity of these immuno-enzymosomes by increasing the enzyme density on the immunoliposomal surface. METHODS: To achieve coupling of GUS to the liposomes, introduction of extra thiol groups was required. Two thiolating agents were examined: iminothiolane and SATA. RESULTS: When iminothiolane was used, aggregation of enzymosomes was observed above enzyme densities of 10 micrograms GUS/mumol lipid (TL). An increased electrostatic repulsion of the enzymosomes, created by inclusion of additional negatively charged lipids and by lowering the ionic strength of the external aqueous medium resulted in enzyme densities > or = 20 micrograms GUS/mumol TL without aggregation. Utilizing SATA, > or = 30 micrograms GUS/mumol TL could be coupled without aggregation, even at physiological ionic strength. It was shown that the enzyme density on immuno enzymosomes, and thus on the tumor cell surface, strongly influences the antitumor effect of the prodrug daunorubicin-glucuronide against in vitro cultured ovarian cancer cells. The antitumor effect of immuno-enzymosomes with enzyme densities of about 20 micrograms GUS/mumol TL was similar to that of the parent drug daunorubicin. CONCLUSIONS: SATA-mediated thiolation of GUS-molecules enabled the preparation of immuno-enzymosomes with high enzyme densities while avoiding spontaneous aggregation. In vitro antitumor activity experiments showed that the improved immuno-enzymosome system is able to completely convert the prodrug daunorubicin-glucuronide into its parent compound. PMID- 8710755 TI - Determination of R(+)- and S(-)-lansoprazole using chiral stationary-phase liquid chromatography and their enantioselective pharmacokinetics in humans. AB - PURPOSE: Stereoselective and sensitive methods employing chiral stationary phase columns for HPLC determination of enantiomers of lansoprazole in the human serum were developed and pharmacokinetic behaviors of the enantiomers were evaluated in seven subjects. METHODS: Five chiral stationary phase columns: Chiralcel OD (cellulose tris(3,5-dimethyl-phenylcarbamate)), OF (cellulose tris(4 chlorophenylcarbamate)), OG (cellulose tris(4-methylphenylcarbamate)) and OJ (cellulose tris(4-methylbenzoate)), and Chiralpak AS (amylose tris ((S)-1 phenylethylcarbamate)) were investigated. RESULTS: Chiralcel OD and Chiralpak AS columns gave a good resolution of R(+)- and S(-)-enantiomers from racemic lansoprazole, but Chiralcel OF, OG, and OJ did not. The mean Cmax and the AUC values of R(+)-enantiomer were 3-5 times greater than those of S(-)-enantiomer following oral administration of 30 mg of racemic lansoprazole. The CLtot values of R(+)-enantiomer were significantly smaller than those of S(-)-enantiomer. Binding of R(+)-enantiomer to human serum proteins was significantly greater than that of S(-)-enantiomer. The mean metabolic ratio (metabolites/parent compound) in human liver microsomes of S(-)-enantiomer was significantly greater than that of R(+)-enantiomer. CONCLUSIONS: The stereoselective pharmacokinetics of lansoprazole enantiomers is likely due to its stereoselective protein binding and/or metabolism. PMID- 8710756 TI - Use of near-infrared spectroscopy to evaluate an active in a film coated tablet. AB - PURPOSE: To provide a method to rapidly screen tablets in the development of new coating technology. METHODS: Near-Infrared (NIR) reflectance spectroscopy was used to quantitatively analyze tablets which were composed of a drug active encasing an active drug core. Diffuse reflectance NIR scans of 240 individual tablets over the range of 1100-2500 nm were obtained. High Performance Liquid Chromatography (HPLC) was used as the reference method. RESULTS: Both qualitative, Principal Component Analysis, and quantitative results showed a strong agreement between the NIR and HPLC methods. The NIR analysis was non invasive and allowed subsequent testing of the tablets. The contents of the drug active contained in a drug coating was determined to +/- 4% of the target value using NIR analysis. Over 400 samples were analyzed in less than a month utilizing this technique which allowed the optimization of a new coating technology. CONCLUSIONS: NIR analysis allowed the evaluation of the efficiency of a new drug film coating manufacturing process more quickly and inexpensively. Because the Near-Infrared method was non-invasive the tablets were available for further analysis unlike the chromatography method. PMID- 8710757 TI - The binding of cyclosporin A to human plasma: an in vitro microdialysis study. AB - PURPOSE: The human plasma binding of cyclosporin A was studied in vitro using the technique of microdialysis. The effect of temperature on the overall binding interaction between cyclosporin A and human plasma was also investigated. METHODS: Flow-through loop-type microdialysis probes were constructed from fused silica tubing and regenerated cellulose tubing with a MWCO of 13000 daltons. Probes were perfused with phosphate buffer (0.5 microliters/min) and the concentration of 3H-cyclosporin A in the well-mixed medium (plasma or buffer) was 1200 ng/ml. Relative recoveries of cyclosporin A from plasma or buffer were determined for each probe by separate experiments to measure the solute gain or loss with reference to the perfusate. RESULTS: Recoveries determined by loss were significantly greater than those determined by gain and in each case temperature dependent, with higher recoveries at higher temperatures. The plasma free fraction of cyclosporin A calculated from the recovery data and the perfusate to plasma concentration ratios was dependent on temperature in a log-linear fashion. Mean +/- s.d. plasma free fractions expressed in percent were 33.5 +/- 4.6, 17.9 +/- 3.6, 6.2 +/- 0.8, 3.0 +/- 0.6, and 1.5 +/- 0.2 at temperatures of 4, 10, 20, 30, and 37 degrees C, respectively. Assuming that the enthalpy of binding is constant over the temperature range studied and pseudo-first order conditions exist, the binding reaction at these temperatures was spontaneous, endothermic (delta H = 74.0 kJ/mole), and entropically driven (delta S = 0.274 kJ/mole/deg). CONCLUSIONS: These results show that the free fraction of cyclosporin A in human plasma is dependent on temperature with the fraction unbound decreasing with temperature in the range of 4 to 37 degrees C. The thermodynamic parameters for the binding of cyclosporin A to plasma components indicate that the reaction is a spontaneous endothermic reaction that is mainly entropy driven, similar to the partitioning of lipophilic molecules from an aqueous to a hydrophobic phase. Moreover, these results show that microdialysis is a feasible method to determine the binding interactions between plasma and cyclosporin A, which indicates the method may be suitable for other difficult binding studies where the solutes have nonspecific binding to separation devices. PMID- 8710758 TI - Pharmacokinetic interaction between intravenous 2',3'-dideoxyinosine and pentamidine in rats. AB - PURPOSE: This study examined the pharmacokinetic interaction between 2',3' dideoxyinosine (ddI) and pentamidine. BACKGROUND: ddI and pentamidine are often coadministered to patients with acquired immunodeficiency syndrome, and are both associated with pancreatic toxicity. Information on potential interaction would be useful to assess the need for dose modification and the basis of the higher incidence of pancreatic toxicity associated with coadministration of the two drugs. METHODS: ddI (200 mg/kg) and pentamidine (10 mg/kg) were administered by continuous infusion to rats over 3 hr, either alone or concomitantly. Drug analysis was by high pressure liquid chromatography with UV or fluorescence detection, or by radioimmunoassay. RESULTS: Pentamidine coadministration significantly increased the apparent volume of distribution at steady state of ddI from 1.4 to 3.4 l/kg (p = 0.004), and increased the mean residence time from 36.3 to 50.0 min (p = 0.015). Pentamidine enhanced the distribution of ddI from plasma into pancreas (p = 0.001) and muscle (p = 0.026). ddI distribution into spleen and liver was also increased, with differences approaching statistical significance (p = 0.08 and 0.06, respectively). In contrast, ddI coadministration did not affect the total body clearance but increased the urinary excretion and the renal clearance of pentamidine by about 5-fold (p = 0.0003). CONCLUSIONS: These data indicate that pentamidine increased the distribution of ddI into pancreas and muscle, whereas ddI increased the renal elimination of pentamidine. PMID- 8710759 TI - Synergistic effect of electric field and ultrasound on transdermal transport. PMID- 8710760 TI - Effects on the pharmacokinetics and pharmacodynamics in the elderly of coadministering ramipril with water, apple juice, and applesauce. PMID- 8710761 TI - The effect of the reconstitution medium on aggregation of lyophilized recombinant interleukin-2 and ribonuclease A. PMID- 8710762 TI - Questions of parental anxiety. PMID- 8710763 TI - Towards informed decisions about prenatal testing: a review. AB - There are now several well-documented psychological problems associated with prenatal testing programmes. These include poor understanding of tests undergone or declined, anxiety following false positive results, and false reassurance in those receiving negative test results. There is, as yet, little evidence concerning how to provide services to circumvent these. The focus of this review is upon just one of these problems: how best to inform women about prenatal testing and their reproductive options following the diagnosis of a fetal abnormality. Possible methods of improving informed decision-making either about whether to undergo testing or whether to terminate an affected pregnancy are described drawing upon research from antenatal and other health care areas. Future challenges for clinical practice and research in this area concern the range of conditions and predispositions for which prenatal testing with the option of termination should be offered. PMID- 8710765 TI - Ultrasound screening for fetal abnormalities. AB - Ultrasound screening for fetal abnormalities is increasingly becoming part of routine antenatal care in Europe and the UK. However, there has been very little formal evaluation of this practice. In this article reports of routine ultrasound screening are reviewed and the advantages and disadvantages discussed. The majority of routine anomaly scanning is done in the second trimester but there may be a case for screening at other times in pregnancy and alternative anomaly screening policies are discussed. PMID- 8710764 TI - First trimester serum screening for Down's syndrome. AB - Screening for Down's Syndrome has been shown to be effective at 10 weeks of pregnancy. A multicentre study (the first trimester serum screening study) has shown that there are two biochemical markers of choice at this time in pregnancy, namely pregnancy associated placental protein A (PAPP-A) and the free beta-sub unit of human chorionic gonadotrophin (free beta-hCG). When used together with maternal age these two biochemical markers have an estimated detection rate of 62% and a 5% false-positive rate. The results are consistent with those obtained from a systematic review of the world literature. Other markers are less predictive of Down's syndrome though there is still some uncertainty over the value of dimeric inhibin-A at 10 weeks of pregnancy. Nuchal translucency measurement, from an ultrasound examination performed at about 10 weeks of pregnancy, is associated with Down's syndrome and is emerging as an important potential screening marker. At present there is uncertainty over its quantitative performance and performance when combined with biochemical markers. The resolution of these issues is currently the subject of active research. Ten week screening for Down's syndrome is an advance that is now technically possible though there is still insufficient information to justify its use in routine screening practice. PMID- 8710766 TI - Early amniocentesis: a clinical review. AB - Early amniocentesis at less than 14 weeks gestation is becoming more common in prenatal diagnosis populations. Randomized studies are minimal and have not had the power to determine the accuracy and safety of the procedure compared to chorionic villus sampling or mid-trimester amniocentesis. Procedures at 11+0-12+6 weeks should be considered experimental. This clinical review considers the ethics, embryology, and clinical experience (cytogenetics, AFP, AChE, procedure and cytogenetic failures, spontaneous and therapeutic pregnancy losses, congenital anomalies) of early amniocentesis. PMID- 8710767 TI - Prenatal screening for haemoglobin disorders. AB - The technology has been available to detect carriers of haemoglobin disorders since the late 1960s. Prenatal diagnosis has been available since 1978. First trimester diagnosis by chorionic villus sampling and DNA analysis was introduced in 1982, and subsequent simplifications in DNA technology have made screening, counselling and prenatal diagnosis cost-effective at the community level, in countries at all levels of development. Audit of prenatal diagnosis for haemoglobin disorders in countries which have the resources and infrastructure necessary for genetic population screening (such as the UK and other European countries), has shown that the number of prenatal diagnoses actually performed fall far short of expectation. The demonstration that this reflects failures in delivering information, screening and counselling to the populations at risk, rather than rejection of prenatal diagnosis, shows the importance of placing more emphasis on the organisational and social requirements for genetic population screening. In some countries current attitudes towards abortion exclude provision of prenatal diagnosis within the health service, but in many such cases it has been set up in the private sector. It is also being introduced through combined private and charitable efforts in an increasing number of developing countries, including some with extremely limited health resources: such centres are likely to act as nuclei for emergence of genetics services in these communities. A particularly notable recent achievement is the introduction of prenatal diagnosis in Nigeria, where 1-2% of all children born suffer from sickling disorders. PMID- 8710768 TI - The efficacy of flecainide versus digoxin in the management of fetal supraventricular tachycardia. AB - Fetal supraventricular tachycardia (SVT) can be successfully treated transplacentally, but in cases where fetal hydrops develops there is considerable morbidity and mortality. The present study was carried out to establish whether the introduction of flecainide altered obstetric management and fetal outcome. A retrospective analysis took place of 51 singleton pregnancies which were referred to the division of prenatal diagnosis because of fetal tachycardia between 1982 and 1993. SVT was documented in 50 out of 51 fetuses, one of which displayed a combination of extensive rhabdomyomas and severe hydrops and died shortly after referral. In the other fetus ventricular tachycardia was diagnosed. Of the remaining 49 fetuses, 14 did not receive any prenatal treatment, but nine needed postnatal treatment. Transplacental treatment of SVT took place in 35 fetuses, of which 22 presented without hydrops and 13 with hydrops. These subsets differed significantly with respect to restoration of normal sinus rhythm (73% vs. 30%; p < 0.001) and mortality (0% vs. 46%; p < 0.001). Digoxin was effective in restoring sinus rhythm in 55 per cent of the non-hydropic fetuses but in only eight per cent of the hydropic fetuses. Flecainide was effective in restoring sinus rhythm in all non-hydropic fetuses where digoxin treatment failed, and in 43 per cent of hydropic fetuses. Administration of flecainide resulted in a significantly reduced mortality (p < 0.001) compared with digoxin treatment. No adverse effects were seen. Postnatal anti-arrhythmic treatment was necessary in 23 infants. Treatment could be withdrawn within one year in all cases but one. PMID- 8710769 TI - Prenatal gene therapy: prospects and issues. PMID- 8710770 TI - Chorionic villus sampling and materno-fetal transfusions: an immunological pathogenesis of vascular disruptive syndromes? AB - Experimental materno-embryonic transfusions with serum that is immunologically active against blood group antigens cause congenital malformations in the rat embryo. In view of the possible increased incidence of vascular disruptive syndromes after chorionic villus sampling (CVS), we investigated the occurrence of materno-fetal transfusions (MFTs) in this procedure. In 18 pregnant women experiencing two needle introductions at CVS, we looked immunohistochemically at the presence of haemoglobin A1-containing maternal erythrocytes in the fetal circulation of the separately collected first and second chorionic villus samples. In 4 of 18 patients (22 per cent), a significant increase of maternal cells was observed in the second sample compared with the first sample, indicating the occurrence of MFT by CVS. On the rare occasion of maternal immunization against fetal antigens, a CVS-associated MFT might provoke immunological damage to the fetus. PMID- 8710771 TI - First-trimester sonographic detection of neurodevelopmental abnormalities in some single-gene disorders. AB - A number of single-gene disorders are detectable early in pregnancy through DNA analysis on chorionic villi. In some genetic disorders that are not yet amenable with this technique, prenatal diagnosis is based on the sonographic detection of neurodevelopmental abnormalities. In nine fetuses at risk for the Meckel-Gruber, Walker-Warburg, and Joubert syndromes, sonographic scanning was accomplished in the first and early second trimesters of pregnancy. In five out of nine fetuses, structural anomalies were detected. In two out of five affected fetuses, sonographic detection of a cephalocele was predated by the visualization of an enlarged rhombencephalon cavity diameter. PMID- 8710772 TI - Early sonographic detection of fetal intestinal obstruction and possible diagnostic pitfalls. AB - We present five fetuses at 15-17 weeks' gestation with a sonographic diagnosis of intestinal obstruction. Dilation of the intestine was the presenting sonographic finding in fetuses with a volvulus and/or anal atresia. Two of the fetuses also had other abnormalities. Intestinal peristalsis may be observed in early pregnancy as transient dilation of intestinal segments. Sonographers are cautioned about a false-positive diagnosis of intestinal obstruction in such cases. PMID- 8710773 TI - Prenatal screening for congenital nephrosis in east Finland: results and impact on the birth prevalence of the disease. AB - Congenital nephrosis of the Finnish type (CNF) is inherited as an autosomal recessive trait which maps to the long arm of chromosome 19. The disease causes massive proteinuria, and renal transplantation in early neonatal life is the only effective treatment. Prenatal diagnosis, usually in high-risk families, depends on the analysis of alpha-fetoprotein (AFP) levels in maternal serum (MS) or amniotic fluid (AF). We studied the effectiveness of MSAFP measurement as a method of screening all pregnant women for congenital nephrosis. Between 1 January 1979 and 31 December 1992, all pregnant women (N = 110,858) attending maternity care units in East Finland were offered serum AFP measurement as part of a screening programme. All patients whose MSAFP value was > or = 2.5 multiples of the median (MOM) at 15-18 weeks' gestation were given an ultrasound examination and if no morphological abnormality was found, they were then offered amniocentesis. Altogether, 105,880 pregnant women (96 per cent) in East Finland participated in the screening for CNF. A total of 47 cases of CNF were diagnosed during the study period. Elevated AFP concentrations in maternal serum and amniotic fluid (> or = 2.5 MOM) were found in all screened (44/105,880) affected pregnancies except one. The most typical feature of a CNF pregnancy was a very high AFP concentration in the amniotic fluid. As a result of the screening, the prevalence at birth of CNF decreased from 1:2600 to 1:11,086. The possibility of CNF has to be taken into account in pregnancies with a ?false-positive' elevated AFP result (normal ultrasound and no detectable acetyl cholinesterase). PMID- 8710774 TI - Prenatal diagnosis of cystic fibrosis in a highly heterogeneous population. AB - Cystic fibrosis (CF) is the most common autosomal recessive disease in Caucasian populations. The Spanish CF population is highly heterogeneous, with more than 70 different mutations causing CF. Since the CFTR gene was cloned, we have performed 81 prenatal diagnoses for 74 couples. Sixty-nine cases had a high risk (1/4) for CF and 12 presented a lower risk (1/240). Direct analysis was possible in 36 cases (44.4 per cent); it was necessary to combine mutation analysis with polymorphic markers in 24 cases (29.6 per cent); mutation analysis and microvillar enzymatic (MVE) analysis were combined in five cases (6.1 per cent); and in 16 cases (19.8 per cent), only indirect analysis was possible. Nine different mutations were detected in this series of families: 621 + 1G -> T, delta F508, 1609delCA, G542X, G551D, 1949del84, R1162X, W1282X, and N1303K. Another ten mutations were identified in these samples after prenatal diagnosis (1811 + 1.6kbA -> G, 711 + 1G -> T, 2869insG, G85E, 2176insC, delta I507, 3272 26A -> G, Q890X, R1066C, and 4005 + 1G -> A). Our current strategy for molecular diagnosis of CF in the Spanish population is based, as a first step, on direct analysis for the two most frequent mutations (delta F508 and G542X) and indirect analysis using the intragenic markers IVS8CA, IVS17BTA, and IVS17BCA. The second step consists of screening for the mutations already associated with the CFTR microsatellite haplotypes. The third step is a specific search for unknown mutations. While actual diagnostic methods are not automatic and robust enough for heterogeneous populations, the diagnostic strategy outlined provides rapid, accurate, and reliable prenatal diagnosis for the majority of couples. PMID- 8710775 TI - Immediate changes in umbilical blood flow after transcervical chorionic villus sampling performed by biopsy forceps. AB - Transvaginal ultrasonography with colour Doppler was used to evaluate the immediate changes on the fetal heart rate (FHR) and the fetal umbilical artery pulsatility index (PI) after chorionic villus sampling (CVS). This prospective study included 279 consecutive singleton pregnancies, between 10 and 13 weeks, of women who underwent transcervical CVS in our institution. All Doppler measurements were obtained transvaginally before and immediately after CVS. Structural malformations detected by ultrasound were excluded. Student's t- and Wilcoxon texts were performed for statistical analysis. The results showed no significant decrease in FHR (mean 1.04 beats, t = 1.68, P = 0.94) and a significant elevation of umbilical artery PI (mean 0.12, t = - 6.51, P < 0.001) post-CVS. This difference was only significant in procedures performed at less than 11 weeks' gestation, since there was no significant change for those procedures performed thereafter. These preliminary data suggest that acute fetal haemodynamic changes are induced by CVS and may have clinical effects. Hypotheses on possible vascular mechanisms are discussed. Colour Doppler could widen the possibilities for more accurate in vivo assessment and research on CVS-related placental injury. PMID- 8710776 TI - Prenatal diagnosis of X-linked myotubular myopathy: strategies using new and tightly linked DNA markers. AB - X-linked myotubular myopathy (MTM1) is a severe congenital myopathy characterized by hypotonia, muscle weakness, and associated respiratory insufficiency. Perinatal death is common. The disease locus was shown to be linked to polymorphic markers in Xq28 and we have recently refined the MTM1 locus to a physical region of less than one megabase (Mb) at proximal Xq28. Two new microsatellite markers were developed and assigned in the MTM1 candidate region. We applied them and other DNA markers for prenatal diagnosis in two families. In one case, an affected fetus was predicted and a recombination event was observed with two more distal markers in the region. The second fetus was born unaffected as predicted. The new DNA markers and the precise location of the MTM1 gene provide an improvement for early prenatal diagnosis of the disease. We present suggestions for different combinations of linked and flanking DNA markers for maximal informativeness and accuracy. PMID- 8710777 TI - A case of tyrosinaemia type I with normal level of succinylacetone in the amniotic fluid. AB - Prenatal diagnosis of tyrosinaemia type I can be achieved in cultured amniotic cells and in chorionic villus material by testing the activity of fumarylacetoacetate hydrolase and by DNA analysis, and in amniotic fluid by succinylacetone measurement. This specific metabolite can be measured either by gas chromatography-mass spectrometry or by delta-aminolevulinate dehydratase inhibition assay. In a series of 65 at-risk cases tested with the enzyme inhibition assay, one case out of the 18 with the disease had a normal level of succinylacetone. This case is presented. PMID- 8710778 TI - Agenesis of the ductus venosus in a case of monochorionic twins which mimics twin twin transfusion syndrome. AB - We describe a pair of monochorionic twins with clinical manifestation of twin twin transfusion syndrome, including discordant fetal growth, hydrops of the larger twin, and the 'stuck twin' phenomenon of the smaller twin. In the hydropic twin, we found agenesis of the ductus venosus and an abnormal umbilical vein draining directly into the inferior vena cava. There was also marked enlargement of the right atrium, representing the volume overload on it. We believe that the agenesis of the ductus venosus in the larger twin may have contributed to the 'unbalanced shunting' between the monochorionic twins. This then led to a clinical picture mimicking that seen in the authentic twin-twin transfusion syndrome. PMID- 8710779 TI - Langer mesomelic dwarfism: ultrasonographic diagnosis of two cases in early mid trimester. AB - This is a report on two cases of mesomelic dysplasia Langer type. This is a rare kind of dwarfism which combines severe dwarfism with predominant shortening of the forearms and lower legs and other morphological abnormalities. The diagnosis can be made by ultrasonography in the early second trimester of the pregnancy. PMID- 8710780 TI - Prenatal diagnosis of beta-thalassaemia and sickle cell anaemia in Turkey. AB - This paper reports our experience of molecular analysis and diagnosis of beta thalassaemia and sickle cell anaemia (HbS) in 70 prospective parents of Turkish descent and their fetuses. Molecular screening was carried out by allele-specific oligonucleotide (ASO) hybridization of amplified DNA to the 12 most common mutations in the Turkish population. By using this approach, we were able to define the mutation in 95 per cent of chromosomes investigated. Genomic sequencing led to the additional detection of three rare mutations: Cd 44 (-C), IVS-I-5 (G-C), and IVS-I-116 (T-G). All diagnoses were successfully accomplished and no misdiagnosis occurred. Consanguineous marriage appears to contribute significantly to the frequency of affected births in Turkey. Out of the 14 homozygous fetuses, six were the result of close consanguinity. This study indicates that fetal diagnosis of beta-thalassaemia and HbS may be obtained in practically all cases, even in a heterogeneous population like the Turkish population, when early methods of fetal sampling are combined with polymerase chain reaction (PCR)-based techniques. Until gene therapy becomes a reality, the only approaches to the control of haemoglobinopathies are prevention and avoidance. The most relevant and common aspects of the programmes, which have been very effective in reducing the birth rate of beta-thalassaemia major in several at-risk areas of the Mediterranean basin, are the continuous educational campaigns directed at the population at large, the voluntary basis, and non directive counselling. The most important challenge for the eradication of the haemoglobinopathies in Turkey is the organization of a nation-wide and comprehensive genetic preventive programme based on DNA technology. PMID- 8710781 TI - Prenatal diagnosis of adrenoleukodystrophy by means of mutation analysis. AB - Prenatal diagnosis of adrenoleukodystrophy (ALD) was performed by means of genetic and biochemical analysis using chorionic villi and amniocytes. The mother was a carrier of an exonic point mutation in the ALD protein gene (2154 C to T) which resulted in the premature formation of a termination codon (Q590STOP) and deletes the Pst I site. Two patients in this family were hemizygotes for this mutation. Pst I digestion of cDNA from chorionic villi revealed that the fetus was a heterozygote for this mutation, and sex determination using the polymerase chain reaction (PCR) indicated female. Lignocerate oxidation in cultured amniocytes was slightly decreased. These findings suggest that the fetus is a female carrier of ALD, and the resultant baby was female. PMID- 8710782 TI - Prenatal diagnosis of X-linked spinal and bulbar muscular atrophy in a Greek family. AB - X-linked spinal and bulbar muscular atrophy (SBMA) is a late-onset motor neuron disorder which is caused by an expansion of the trinucleotide repeat (CAG)n in the first exon of the androgen receptor gene. Two cases of prenatal testing for the disease in a Greek family are reported. An affected male died in his late 50s of this disorder and his 30-year-old daughter (an obligate carrier) asked for prenatal testing for SBMA. DNA analysis revealed that she indeed carried an expanded allele of 40 repeats, as well as a normal size allele of 24 repeats. Prenatal diagnosis of SBMA was performed when, on two successive pregnancies, two male fetuses with the expanded (CAG)n allele were found. PMID- 8710783 TI - Prenatal diagnosis of Rambam-Hasharon syndrome. AB - Rambam-Hasharon syndrome (RHS) is a newly recognized autosomal recessive inborn error in fucose metabolism. Mental retardation, short stature, coarse facies, and recurrent infections are the main clinical findings. Several fucosilated proteoglycans are deficient in these patients. Leukocyte adhesion deficiency type 2 is associated with lack of the membrane glycoprotein sialyl-Lewisx (CD15s). In the red blood cells (RBCs), lack of the membrane glycoprotein H is manifested as a Bombay (Oh) blood type. Two consecutive pregnancies it risk for RHS were monitored during mid-trimester by cordocentesis. One fetus expressed H substance and her blood phenotype was O Rh+. The second fetus, a female, was 2 weeks smaller than expected by dates and had the Bombay blood type. The placenta of the affected fetus was small and irregular. This is the first prenatal diagnosis of this syndrome and the first case found in a female. The documentation of the syndrome in patients of both sexes and the parental consanguinity support an autosomal recessive inheritance. Two apparent recombinations between fucosyl transferase 1 (FUT1, the H gene) and fucosyl-transferase 2 (secretor) are suggestive of non-allelic heterogeneity. We believe that the Bombay phenotype in this family is caused by a mutated gene, other than FUT1, which is causing multiple deficiencies of fucosilated proteoglycans. PMID- 8710784 TI - Prenatal diagnosis of partial monosomy 3p and partial trisomy 2p in a fetus associated with shortening of the long bones and a single umbilical artery. AB - The prenatal and postnatal findings of a fetus with partial deletion of 3p25 -> pter and duplication of 2p25.3 -> pter are described. The proband postnatally displayed mental and growth retardation, psychomotor delay, microcephaly, ptosis, micrognathia, a narrow palate, and cryptorchidism. All of these anomalies were consistent with those described in 3p- and partial trisomy 2p syndromes, and also frequently seen in patients with other chromosomal disorders. However, the prenatal sonograms revealed unusual shortening of the long bones, a single umbilical artery, and normal development of the skull. Our case suggests that skeletal growth retardation of the long bones may occur earlier than that of the skull in fetuses associated with chromosomal aberrations such as del(3p)/dup(2p). Shortening of the long bones and a single umbilical artery together with other abnormalities detected by prenatal ultrasound thus warrant a fetal cytogenetic study. PMID- 8710786 TI - Current awareness in prenatal diagnosis. PMID- 8710785 TI - Prenatal sonographic diagnosis of mid shaft hypospadias. AB - We report the prenatal diagnosis of mid shaft hypospadias and describe the sonographic features of fetal hypospadias including an abnormal urethral canal, ventral curvature of the distal penis, extension of the penile glans beyond the prepuce, and fetal micturation in a plane perpendicular to the penile shaft. An accurate family history is an essential part of the evaluation of the milder degrees of fetal hypospadias. PMID- 8710787 TI - [How unpolitical may in current times pathology actually be?]. PMID- 8710788 TI - [Tumor suppressor gene p53. Theoretical principles and their significance for pathology]. AB - Mutations of the p53 tumor-suppressor gene are the most common genetic alterations in human cancer, found in approximately 50% of all tumors. The importance of p53 in human cancer attracts attention in molecular studies dealing with the pathogenesis, diagnosis and prognosis in tumor pathology. This review summarizes the current understanding of p53 both on the genetic and protein level. Frequency and spectrum of somatic p53 mutations in the carcinogenesis of breast cancer, colorectal cancer, gastric cancer, hepatocellular carcinoma, squamous-cell carcinoma of the skin and malignant melanoma are discussed including our own investigations and studies published in the literature. PMID- 8710789 TI - [Comparative genomic hybridization in pathology. A new molecular cytogenetic method]. AB - Comparative genomic hybridisation (CGH) is a new cytogenetic method, which is based on a combination of fluorescence microscopy and digital image analysis. The molecular genetic basis is the hybridization of a mixture of fluorescein labeled test-DNA and reference-DNA on normal metaphase chromosomes. Comparative analysis allows the identification of all unbalanced chromosomal aberrations of the test DNA in a single experimental step. The resulting DNA gains or DNA losses on the chromosomal or subchromosomal level mirror possible amplifications of oncogenes or losses of suppress orgenes. As CGH can be performed with genomic DNA of formalin-fixed and fresh-frozen tissue or cells, this new method is a very effective tool for pathologists and cytologists in the extended genomic screening of tumors and genetically altered tissues. Despite CGH analysis at present is restricted to research applications; its widespread dissemination as a routine method in diagnostic pathology can be expected in the near future. PMID- 8710790 TI - [CD34 detection--an immunohistochemical contribution to differential diagnosis of soft tissue tumors]. AB - CD34 is a myeloid progenitor cel antigen and present in endothelial cells and almost all vascular lesions. Additionally, CD34 has been described in numerous fibroblast-like cells and different mesenchymal tumours. The immunohistochemical evidence of CD34, however, may be of importance in the differential diagnosis of benign and malignant soft tissue lesions, if clinicopathological features and other immunohistochemical markers are regarded properly. We studied the CD34 immunopositivity of solitary fibrous tumour at different sites (11 out of 11 tumours tested stained positive), dermatofibrosarcoma protuberans (15 out of 18 stained positive), and gastrointestinal stromal tumour (15 out of 15 stained positive), and discuss the significance of these results in differential diagnosis to morphologically comparable soft tissue lesions. PMID- 8710791 TI - [C-erbB-2, PCNA and histomorphologic factors in breast carcinoma after oral contraceptive use]. AB - The influence of oral contraceptives (OC) on histomorphological and molecular biological prognostic factors was studied in 471 breast cancer patients. Differences in histological tumor type, histological grade, tumor size, lymph node status, hormonal receptor status, PCNA expression and c-erbB-2 protein overexpression were investigated in relation to the duration of OC use (< 5 years/ > or = 5 years) and the time since last use. A total of 297 (63%) patients had used oral contraceptives at some time in their life; 186 patients (39.5%) had used OC's for 5 years or more. There were no significant differences in the tumor characteristics investigated with respect to OC use in general. Neither long-term use at some time in their life nor long-term use until breast cancer diagnosis had an effect on histomorphological and molecular biological factors. Thus, steroid hormones contained in OC's had no direct effect on prognostic factors in breast cancer. PMID- 8710792 TI - [Mucin histochemical analysis of mucin-forming carcinomas, metaplasias of the urothelium and in persistent urachus]. AB - Twelve cases of mucus secreting metaplasias, nine mucus secreting urothelial carcinomas and three cases of urachus remnants were studied using D-PAS-, m-PAS,- AB pH 2,5/PAS-, HID/AB pH 2,5- and PB/KOH/PAS-technique. Neutral mucins, sialomucins, sulfomucins, O-acylated and unsubstituted or C4- and C7-substituted mucins were detected in different quantities, often intracellular mucus mixtures could be seen. Many differences in the mucus quality of the metaplasias, in comparison to that of the tumors, were not noticeable. Metaplasia associated with carcinoma and mucus secreting carcinomas showed a loss of O-acylated sialomucins and besides diffus positive reaction in m-PAS-technique. Further investigations should confirm if this is really a prognostic factor. O-acylated sialomucins were not only found in metaplastic lesions and urothelial carcinomas but also in metaplasias of the urachus. Therefore it is not possible to distinguish between adenocarcinomas of the urinary bladder and adenocarcinomas derived from the Urachus using PB/KOH/PAS method. PMID- 8710793 TI - [Giant axonal neuropathy. A case report]. AB - We report a sporadic case of giant axonal neuropathy (GAN) starting in a 5-year old boy with gait disturbance. At the age of 10 years he showed signs of a sensomotoric polyneuropathy and central neurological symptoms: intention tremor, nystagmus and visual disturbance. The boy also had curly hair. Sural nerve biopsy showed many giant axons with accumulation of microfilaments, which were also found in cytoplasma of Schwann cells, endothelial cells and fibrocytes. Similar changes were detected in endothelial cells of a muscle biopsy, which additionally displayed neurogenic atrophy and target fibers. Immunohistochemically, desmin, vimentin, heat shock protein 70, and ubiquitin were not detectable in giant axons, which, however, contained neurofilament protein 68, 200 and beta-tubulin. Our case shows that careful ultrastructural examination of a muscle biopsy may point to the diagnosis of GAN. PMID- 8710794 TI - [Lung metastasis of a benign giant cell tumor of the skeleton 27 years after resection of a recurrent tumor]. AB - Benign giant cell bone tumors sometimes develop benign pulmonary metastases. Most of these appear 3 to 4 years after the treatment of the primary tumor. Metastases have been diagnosed up to 11 years after the first diagnosis. The following is a case report of a 58-year-old man who developed lung metastases 27 years after the treatment of an aggressive local recurrence. PMID- 8710795 TI - [Primary multiple malignant schwannoma of the heart--a rarity]. AB - The case is presented of a 51-year-old woman with increased of nightly dyspnea. An echocardiographic examination shows two intracardiac tumors. Histological features demonstrate an malignant schwannoma. The diagnosis was verified by immunohistological proof of S-100 protein in tumor cells. PMID- 8710796 TI - [Pathologic-anatomic examination of prostatovesiculectomy samples in prostate carcinoma]. AB - The incidence of the prostatic carcinoma as well as the cases treatable by radical prostatovesiculectomy with curative intention is increasing. The exact pathological examination of the specimens defines the stage of disease and subsequently the prognosis and therapeutic consequences. The compatibility of different scientific studies is made difficult by the lack of a standard in the pathological examination of the specimens. Our exactly defined rational method helps eliminate these troubles and ameliorate the interdisciplinary dialogue. PMID- 8710797 TI - ["Walking pneumonia" in primary sarcoma of the pulmonary artery]. AB - A 74-year-old man suffered from symptoms of a left upper lobe pneumonia. Additional retrosternal masses were proved. Progression in the left upper lobe occurred in spite of antibiotic therapy; infiltrations of the right lower lobe were seen. No germs could be identified. Bronchoscopy and mediastinoscopy showed no pathological findings. His general health became worse, and the patient died under the clinical diagnosis of "wandering pneumonia" with finally suspicion of fulminant pulmonary artery embolism. Autopsy findings revealed an advanced stage of pulmonary artery sarcoma with myocardial infiltration and metastases in both kidneys and thyroid gland. General manifestation of pulmonary artery sarcoma usually are symptoms similar to those caused by pulmonary artery embolism with obstruction of pulmonary arterial flow. The unusual constellation of clinical findings similar to colliquating pneumonia results in secondary infection due to tumorous pulmonary infarction. The difficulties in diagnosis by biopsy are caused by primary intravascular growth of the neoplasms. Therefore early diagnosis in an operable tumor stage by means of CT, MR, and angiography is very important. PMID- 8710798 TI - Effects of variation in oxygen tension on responses of the human fetoplacental vasculature to vasoactive agents in vitro. AB - The human placenta perfused in vitro with Krebs' solution has been used to examine the effects of low oxygen tension on the vasoreactivity of the fetal placental vessels to several vasodilator and vasocontrictor autacoids. Increases in fetal arterial perfusion pressure (FAP) produced by endothelin-1 (ET-1, human), the thromboxane A2-mimetic U46619, 5-hydroxytryptamine (5-HT), angiotensin II (A II) and bradykinin (BK) were examined under conditions of high ( >or= 450 mmHg) and low 0.5 microM) results in a further 2-2.5 fold activation of NO synthesis, with half-maximal stimulation observed at 26 +/- 8.2 microM BH4 (mean +/- SEM, n = 4). Chelation of Ca2+ in the medium abolishes the stimulatory effect, indicating that only a Ca2(+)-dependent NO-synthase (NOS) isoform is activated by BH4. Based on our previous findings, we suggest that this isoform is the endothelial or Type III NOS. Importantly, BH4 has no significant effect on the Ca2(+)-dependency of NOS activity, the apparent Km values for Ca2+ are comparable in the absence (1.8 +/- 0.4 microM, mean +/- SEM, n = 6) or presence (2.5 +/- 0.6 microM, mean +/- SEM, n = 6) of 50 microM BH4. The BH4 content of these placentae is 207.4 +/- 86.7 pmol/g wet tissue (mean +/- s.d., n = 9), therefore, BH4 added to the homogenate does not simply restore the concentrations that occur endogenously. The results provide the first evidence that in the early human placenta, a constitutively expressed CA 2(+)-dependent NOS isoform is stimulated by exogenous BH4, raising the possibility that BH4 is an important regulator of NOS activity in this tissue. This novel aspect of the NO-generating pathway may have implications in the aetiology and treatment of pregnancy-induced hypertension and pre-eclampsia. PMID- 8710816 TI - Functional specialization in the ruminant placenta: evidence for two populations of fetal binucleate cells of different selective synthetic capacity. AB - Trophoblast binucleate cells (BNC) in the ruminant placenta demonstrate a characteristic development, mature structure and migratory capacity whether situated in cotyledonary or intercotyledonary regions of the placenta. However, previous immunocytochemical studies demonstrated clear differences in gene expression in granule contents of BNC according to their anatomical location with some proteins being expressed in all BNC (e.g. ovine placental lactogen) whereas others were unique to a particular origin (e.g. SBU3 antigen in cotyledonary BNC only). We have used enriched preparations of binucleate cells and showed differences in steroid metabolic capacity in vitro which is more related to their species origin (sheep or goat) than to their anatomical location. The predominant product from [3H]pregnenolone is progesterone (sheep) and 5 beta-pregnane-3 alpha, 20 alpha-diol (goat) and the amount formed (corrected for the number of BNC) is similar irrespective of whether BNC were derived from the cotyledonary or intercotyledonary regions. These studies indicate specific forms of regional functional specialization of BNC and emphasize their multifunctional role in the ruminant placenta. PMID- 8710817 TI - [Lung scintigraphy and ergospirometry in prediction of postoperative course in lung resection candidates with increased risk of postoperative complications]. AB - Patients with impaired pulmonary function are at increased risk for the development of postoperative complications. We therefore analyzed the value of preoperative lung scanning and exercise testing for the prediction of postoperative complications and of the short- as well as long-term performance in lung resection candidates at increased risk for complications. Twenty-five (mean age 63 y; 17 m) out of 84 consecutive lung resection candidates were considered at increased risk for postoperative complications due to impaired pulmonary function (FEV1 < 2L or diffusion for carbon monoxide (DLCO) < 50% predicted, or FEV1 and DLCO < or = 80% predicted combined with New York Heart Association dyspnea index > or = 2). Candidates underwent radionuclide perfusion scans and exercise testing to predict postoperative ( = ppo) values for FEV1, DLCO and maximal O2-uptake (VO2max). They all underwent thoracotomy for neoplastic lesions; 7 had pneumonectomies, 18 lobectomies. Six had postoperative complications (within 30 days), of whom three died. Three and 6 months postoperatively, pulmonary function tests and VO2max were repeated. In the 22 survivors, the observed values were then compared with the predicted values. At 3 months, there were excellent correlations (absolute/predicted values): for FEV1r = 0.78 and 0.81; for DLCOr = 0.77 and 0.74; and for VO2max r = 0.71 and 0.83. The means of FEV1 and VO2max did not differ from the predicted values, whereas the predicted DLCO was lower than the observed value (ml/min/mmHg: 15.1 vs 17.9; percent predicted: 59.6 vs 70.9) (p < 0.05). At 6 months, correlations remained very good for FEV1 (r = 0.81 and 0.84) and for DLCO (r = 0.76 and 0.74), but had decreased for VO2max to 0.56 and 0.65, respectively. All means were higher than predicted (p < 0.05) owing to recovery in the lobectomy group. Patients with postoperative complications (group B) had a lower preoperative VO2max in percent predicted (62.8 +/- 7.5% vs 84.6 +/- 19.7%) (p < 0.01) and also a lower VO2max ppo (10.6 +/- 3.6 vs 14.8 +/- 3.5 ml/kg/min and 44.3 +/- 13.5 vs 68.0 +/- 20.7 percent predicted) (p < 0.05) than patients without complication (group A). AVO2max-ppo < 10/ml/kg/min was associated with a 100% mortality. Although FEV1 ppo and DLco-ppo were lower in group B the difference did not reach significance. We conclude that radionuclide-based calculations of postoperative VO2max are predictive of perioperative morbidity and mortality: a VO2max-ppo of < 10 ml/kg/min may indicate inoperability. Further, short-term postoperative performance is accurately predicted by FEV1-ppo and VO2max-ppo, but long-term function is underestimated after lobectomy. PMID- 8710818 TI - [Respiratory and cardiovascular effects of acetylcholine provocation after inhalation exposure to various occupational pollutants--studies in the rabbit]. AB - Airway hyperresponsiveness, manifested by increased flow resistance and resulting drop in oxygen partial pressure when conducting provocation tests, is considered an early sign of a developing obstructive airway disease, an example of which is a professionally conditioned asthma. We conducted a detailed study exploring the interrelation between the respiratory mechanical parameters and the partial pressures of oxygen and carbon dioxide (PaO2, PaCO2). Reproducibility tests for the studied cardiovascular, ventilatory and respiratory mechanical parameters at rest and under various conditions of stress (external stenoses, inhalation of hypercapnic and hypoxic gas mixtures, infusion of an acetylcholine solution) showed good reproducibility of the measured data with variation coefficients < 10%. In blood gas analyses we also found comparable variation coefficients. Four groups of experimental animals were exposed for different periods of time to various working place substances (coolants, ammonium peroxodisulfate, hair bleaches [blondizing agents], isocyanates). After the exposure we checked on the bronchial sensitivity to aerosols of 0.2% and 2% acetylcholine solutions. Concomitant with an increased response of dynamic elastance, we found an increased drop in oxygen partial pressure and an almost constant carbon dioxide partial pressure, dependent on the working place substance used and on its concentration. In untreated controls the inhalation of acetylcholine resulted in bronchoconstriction and drop in oxygen partial pressure only on provocation with 2% acetylcholine. However, in the groups exposed to coolants and ammonium peroxodisulfate there was a significant drop in oxygen partial pressure already on provocation with 0.2% acetylcholine, as well as a noticeable bronchial respiratory response. The drop in oxygen partial pressure increases further after provocation with 2% acetylcholine, whereas the oxygen partial pressure dropped to a maximum of one-third of its original level by more than 10%. Placing the drop in oxygen partial pressure provoked by acetylcholine in relation to the increase in dynamic elastance, this can be well expressed by a logarithmic formula (y = 6.2. In (x) + 0.72, r = 0.96) that does not change significantly after exposure to working place substances (y = -7.0. In (x) + 3.33, r = 0.93). The close correlation of both parameters suggests that study of the oxygen partial pressures to determine the airway hyperresponsiveness should be considered important, since a marked drop in oxygen partial pressure is seen even if obstructive respiratory response is only slightly increased (slight increase in dynamic elastance). The reason for the behaviour of the blood gases is probably an increased ventilation-perfusion imbalance due to inhomogenous peripheral bronchial reactions. In the hyperresponsive animals the reactions were merely enhanced without demonstrating any differences. PMID- 8710819 TI - [Occupational asthma: definition--epidemiology--etiologic substances--prognosis- prevention--diagnosis--expert assessment aspects]. PMID- 8710820 TI - [Bronchial asthma--a cost of illness analysis]. AB - We performed an economic evaluation of the costs of asthma in Germany. Estimates of direct medical expenditures and indirect costs were derived from official health statistics of 1992. Adding up direct and indirect costs, the total sum is approximately 2.66 billions ECU which is equivalent to 5.13 billions DM or 3.11 billions US-$, respectively, for a total population of 80.3 millions. Direct costs make up 61.5% of this sum and comprise of outpatient medical care, drugs, hospital treatment, rehabilitation and compensation for occupational asthma as well as sickness benefits. Indirect costs of 38.5% are caused by payments for days off work, premature retirement and premature death due to asthma. Based on these figures, we speculate that an optimised treatment according to published recommendations should be accompanied by a reduction of the high amount of indirect costs. PMID- 8710821 TI - [Cardiopulmonary involvement in thyroid gland diseases]. AB - Thyroid hormones exercise a direct influence on the lung and myocardium. It has been found that at the heart of patients suffering from hyperthyroidism there is an increased production of contractile proteins and a redistribution of myosin Isoenzymes. Reduced efficiency results from the increased oxygen consumption and insufficient adaptation of the heartbeat to stress. Thyroid hormones are responsible for reducing arterial resistance; a higher incidence of supraventricular arrhythmias has been observed. In contrast, the cardiac changes in hypothyroidism are associated with an enhanced deposition of glycosaminoglycan in the myocardium. Besides bradycardia and low voltage there is an increased incidence of pericardial, pleural and peritoneal effusions with high protein content. Patients suffering from hypothyroidism have a higher incidence of atherosclerotic changes. Hypothyroidism is associated with a reduction of the beta-adrenergic receptors, and hence disturbances of rhythm are rare and do not increase even under substitution therapy. Pulmonary involvement in hyperthyroidism is evident from an enhanced respiratory minute volume, a reduced vital capacity, and dyspnoea at rest. In patients suffering from hypothyroidism, hypoventilation reduces the efficiency. Over and above this, myopathic changes of the respiratory muscles have been described. Involvement of thyroid hormones in the maturing of type II pneumocytes and of the surfactant factor has also been under discussion. PMID- 8710822 TI - A large-scale experiment to assess protein structure prediction methods. PMID- 8710823 TI - Progress of 1D protein structure prediction at last. AB - Accuracy of predicting protein secondary structure and solvent accessibility from sequence information has been improved significantly by using information contained in multiple sequence alignments as input to a neural network system. For the Asilomar meeting, predictions for 13 proteins were generated automatically using the publicly available prediction method PHD. The results confirm the estimate of 72% three-state prediction accuracy. The fairly accurate predictions of secondary structure segments made the tool useful as a starting point for modeling of higher dimensional aspects of protein structure. PMID- 8710824 TI - A critical assessment of comparative molecular modeling of tertiary structures of proteins. AB - In spite of the tremendous increase in the rate at which protein structures are being determined, there is still an enormous gap between the numbers of known DNA derived sequences and the numbers of three-dimensional structures. In order to shed light on the biological functions of the molecules, researchers often resort to comparative molecular modeling. Earlier work has shown that when the sequence alignment is in error, then the comparative model is guaranteed to be wrong. In addition, loops, the sites of insertions and deletions in families of homologous proteins, are exceedingly difficult to model. Thus, many of the current problems in comparative molecular modeling are minor versions of the global protein folding problem. In order to assess objectively the current state of comparative molecular modeling, 13 groups submitted blind predictions of seven different proteins of undisclosed tertiary structure. This assessment shows that where sequence identity between the target and the template structure is high (> 70%), comparative molecular modeling is highly successful. On the other hand, automated modeling techniques and sophisticated energy minimization methods fail to improve upon the starting structures when the sequence identity is low (approximately 30%). Based on these results it appears that insertions and deletions are still major problems. Successfully deducing the correct sequence alignment when the local similarity is low is still difficult. We suggest some minimal testing of submitted coordinates that should be required of authors before papers on comparative molecular modeling are accepted for publication in journals. PMID- 8710825 TI - Evaluation of comparative protein modeling by MODELLER. AB - We evaluate 3D models of human nucleoside diphosphate kinase, mouse cellular retinoic acid binding protein I, and human eosinophil neurotoxin that were calculated by MODELLER, a program for comparative protein modeling by satisfaction of spatial restraints. The models have good stereochemistry and are at least as similar to the crystallographic structures as the closest template structures. The largest errors occur in the regions that were not aligned correctly or where the template structures are not similar to the correct structure. These regions correspond predominantly to exposed loops, insertions of any length, and non-conserved side chains. When a template structure with more than 40% sequence identity to the target protein is available, the model is likely to have about 90% of the mainchain atoms modeled with an rms deviation from the X-ray structure of approximately 1 A, in large part because the templates are likely to be that similar to the X-ray structure of the target. This rms deviation is comparable to the overall differences between refined NMR and X-ray crystallography structures of the same protein. PMID- 8710826 TI - Confronting the problem of interconnected structural changes in the comparative modeling of proteins. AB - Comparative models of three proteins have been built using a variety of computational methods, heavily supplemented by visual inspection. We consider the accuracy obtained to be worse than expected. A careful analysis of the models shows that a major reason for the poor results is the interconnectedness of the structural differences between the target proteins and the template structures they were modeled from. Side chain conformations are often determined by details of the structure remote in the sequence, and can be influenced by relatively small main chain changes. Almost all of the regions of substantial main chain conformational change interact with at least one other such region, so that they often cannot be modeled independently. Visual inspection is sometimes effective in correcting errors in sequence alignment and in spotting when an alternative template structure is more appropriate. We expect some improvements in the near future through the development of structure-based sequence alignment tools, side chain interconnectedness rotamer choice algorithms, and a better understanding of the context sensitivity of conformational features. PMID- 8710827 TI - Protein structure prediction by threading methods: evaluation of current techniques. AB - This paper evaluates the results of a protein structure prediction contest. The predictions were made using threading procedures, which employ techniques for aligning sequences with 3D structures to select the correct fold of a given sequence from a set of alternatives. Nine different teams submitted 86 predictions, on a total of 21 target proteins with little or no sequence homology to proteins of known structure. The 3D structures of these proteins were newly determined by experimental methods, but not yet published or otherwise available to the predictors. The predictions, made from the amino acid sequence alone, thus represent a genuine test of the current performance of threading methods. Only a subset of all the predictions is evaluated here. It corresponds to the 44 predictions submitted for the 11 target proteins seen to adopt known folds. The predictions for the remaining 10 proteins were not analyzed, although weak similarities with known folds may also exist in these proteins. We find that threading methods are capable of identifying the correct fold in many cases, but not reliably enough as yet. Every team predicts correctly a different set of targets, with virtually all targets predicted correctly by at least one team. Also, common folds such as TIM barrels are recognized more readily than folds with only a few known examples. However, quite surprisingly, the quality of the sequence-structure alignments, corresponding to correctly recognized folds, is generally very poor, as judged by comparison with the corresponding 3D structure alignments. Thus, threading can presently not be relied upon to derive a detailed 3D model from the amino acid sequence. This raises a very intriguing question: how is fold recognition achieved? Our analysis suggests that it may be achieved because threading procedures maximize hydrophobic interactions in the protein core, and are reasonably good at recognizing local secondary structure. PMID- 8710828 TI - Threading a database of protein cores. AB - We present an analysis of 10 blind predictions prepared for a recent conference, "Critical Assessment of Techniques for Protein Structure Prediction." The sequences of these proteins are not detectably similar to those of any protein in the structure database then available, but we attempted, by a threading method, to recognize similarity to known domain folds. Four of the 10 proteins, as we subsequently learned, do indeed show significant similarity to then-known structures. For 2 of these proteins the predictions were accurate, in the sense that a similar structure was at or near the top of the list of threading scores, and the threading alignment agreed well with the corresponding structural alignment. For the best predicted model mean alignment error relative to the optimal structural alignment was 2.7 residues, arising entirely from small "register shifts" of strands or helices. In the analysis we attempt to identify factors responsible for these successes and failures. Since our threading method does not use gap penalties, we may readily distinguish between errors arising from our prior definition of the "cores" of known structures and errors arising from inherent limitations in the threading potential. It would appear from the results that successful substructure recognition depends most critically on accurate definition of the "fold" of a database protein. This definition must correctly delineate substructures that are, and are not, likely to be conserved during protein evolution. PMID- 8710829 TI - Assessment of a protein fold recognition method that takes into account four physicochemical properties: side-chain packing, solvation, hydrogen-bonding, and local conformation. AB - A protein fold recognition method was tested by the blind prediction of the structures of a set of proteins. The method evaluates the compatibility of an amino acid sequence with a three-dimensional structure using the four evaluation functions: side-chain packing, solvation, hydrogen-bonding, and local conformation functions. The structures of 14 proteins containing 19 sequences were predicted. The predictions were compared with the experimental structures. The experimental results showed that 9 of the 19 target sequences have known folds or portions of known folds. Among them, the folds of Klebsiella aerogenes urease beta subunit (KAUB) and pyruvate phosphate dikinase domain 4 (PPDK4) were successfully recognized; our method predicted that KAUB and PPDK4 would adopt the folds of macromomycin (Ig-fold) and phosphoribosylanthranilate isomerase:indoleglycerol-phosphate synthase (TIM barrel), respectively, and the experimental structure revealed that they actually adopt the predicted folds. The predictions for the other targets were not successful, but they often gave secondary structural patterns similar to those of the experimental structures. PMID- 8710830 TI - Progress in fold recognition. AB - The prediction experiment reveals that fold recognition has become a powerful tool in structural biology. We applied our fold recognition technique to 13 target sequences. In two cases, replication terminating protein and prosequence of subtilisin, the predicted structures are very similar to the experimentally determined folds. For the first time, in a public blind test, the unknown structures of proteins have been predicted ahead of experiment to an accuracy approaching molecular detail. In two other cases the approximate folds have been predicted correctly. According to the assessors there were 12 recognizable folds among the target proteins. In our postprediction analysis we find that in 7 cases our fold recognition technique is successful. In several of the remaining cases the predicted folds have interesting features in common with the experimental results. We present our procedure, discuss the results, and comment on several fundamental and technical problems encountered in fold recognition. PMID- 8710831 TI - Successful protein fold recognition by optimal sequence threading validated by rigorous blind testing. AB - Analysis of the results of the recent protein structure prediction experiment for our method shows that we achieved a high level of success. Of the 18 available prediction targets of known structure, the assessors have identified 11 chains which either entirely match a previously known fold, or which partially match a substantial region of a known fold. Of these 11 chains, we made predictions for 9, and correctly assigned the folds in 5 cases. We have also identified a further 2 chains which also partially match known folds, and both of these were correctly predicted. The success rate for our method under blind testing is therefore 7 out of 11 chains. A further 2 folds could have easily been recognized but failed due to either overzealous filtering of potential matches, or to simple human error on our part. One of the two targets for which we did not submit a prediction, prosubtilisin, would not have been recognized by our usual criteria, but even in this case, it is possible that a correct prediction could have been made by considering a combination of pairwise energy and solvation energy Z-scores. Inspection of the threading alignments for the (alpha beta)8 barrels provides clues as to how fold recognition by threading works, in that these folds are recognized by parts rather than as a whole. The prospects for developing sequence threading technology further is discussed. PMID- 8710832 TI - Fold recognition and ab initio structure predictions using hidden Markov models and beta-strand pair potentials. AB - Protein structure predictions were submitted for 9 of the target sequences in the competition that ran during 1994. Targets sequences were selected that had no known homology with any sequence of known structure and were members of a reasonably sized family of related but divergent sequences. The objective was either to recognize a compatible fold for the target sequence in the database of known structures or to predict ab initio its rough 3D topology. The main tools used were Hidden Markov models (HMM) for fold recognition, a beta-strand pair potential to predict beta-sheet topology, and the PHD server for secondary structure prediction. Compatible folds were correctly identified in a number of cases and the beta-strand pair potential was shown to be a useful tool for ab initio topology prediction. PMID- 8710833 TI - Homology modeling by the ICM method. AB - Five models have been built by the ICM method for the Comparative Modeling section of the Meeting on the Critical Assessment of Techniques for Protein Structure Prediction. The targets have homologous proteins with known three dimensional structure with sequence identity ranging from 25 to 77%. After alignment of the target sequence with the related three-dimensional structure, the modeling procedure consists of two subproblems: side-chain prediction and loop prediction. The ICM method approaches these problems with the following steps: (1) a starting model is created based on the homologous structure with the conserved portion fixed and the nonconserved portion having standard covalent geometry and free torsion angles; (2) the Biased Probability Monte Carlo (BPMC) procedure is applied to search the subspaces of either all the nonconservative side-chain torsion angles or torsion angles in a loop backbone and surrounding side chains. A special algorithm was designed to generate low-energy loop deformations. The BPMC procedure globally optimizes the energy function consisting of ECEPP/3 and solvation energy terms. Comparison of the predictions with the NMR or crystallographic solutions reveals a high proportion of correctly predicted side chains. The loops were not correctly predicted because imprinted distortions of the backbone increased the energy of the near-native conformation and thus made the solution unrecognizable. Interestingly, the energy terms were found to be reliable and the sampling of conformational space sufficient. The implications of this finding for the strategies of future comparative modeling are discussed. PMID- 8710834 TI - The use of position-specific rotamers in model building by homology. AB - In this study we concentrate on replacing side chains as a subtask of model building by homology. Two problems arise. How to determine potential low energy rotamers? And how to avoid the combinatorial explosion that results from the combination of many residues for which multiple good rotamers are predicted? We attempt to solve these problems by choosing position-specific rather than generalized rotamers and by sorting the residues that have to be modelled as a function of their freedom in rotamer space. The practical advantages of our method are the quality of the models for cases of high backbone similarity, the small amount of human intervention needed, and the fact that the method automatically estimates the reliability with which each residue has been modeled. Other methods described in this issue are probably more suitable if large backbone rearrangements or loop insertions and deletions need to be modeled. PMID- 8710835 TI - Homology modeling of histidine-containing phosphocarrier protein and eosinophil derived neurotoxin: construction of models and comparison with experiment. AB - Homology modeling methods have been used to construct models of two proteins--the histidine-containing phosphocarrier protein (HPr) from Mycoplasma capricolum and human eosinophil-derived neurotoxin (EDN). Comparison of the models with the subsequently determined X-ray crystal structures indicates that the core regions of both proteins are reasonably well reproduced, although the template structures are closer to the X-ray structures in these regions--possible enhancements are discussed. The conformations of most of the side chains in the core of HPr are well reproduced in the modeled structure. As expected, the conformations of surface side chains in this protein differ significantly from the X-ray structure. The loop regions of EDN were incorrectly modeled--reasons for this and possible enhancements are discussed. PMID- 8710836 TI - Evaluation of current techniques for ab initio protein structure prediction. AB - The results of a protein structure prediction contest are reviewed. Twelve different groups entered predictions on 14 proteins of known sequence whose structures had been determined but not yet disseminated to the scientific community. Thus, these represent true tests of the current state of structure prediction methodologies. From this work, it is clear that accurate tertiary structure prediction is not yet possible. However, protein fold and motif prediction are possible when the motif is recognizable similar to another known structure. Internal symmetry and the information inherent in an aligned family of homologous sequences facilitate predictive efforts. Novel folds remain a major challenge for prediction efforts. PMID- 8710837 TI - The phospho-beta-galactosidase and synaptotagmin predictions. AB - Two bona fide consensus predictions of secondary and tertiary structure in a protein family, made and announced before experimental structures were known, are evaluated in light of the subsequently determined experimental structures. The first, for phospho-beta-galactosidase, identified the core strands of an 8-fold alpha-beta barrel, and identified the 8-fold alpha-beta barrel itself, which was found in the subsequently determined experimental structure to be the core folding domain. The second, for synaptotagmin, identified seven out of eight beta strands in the structure correctly, missing only a noncore strand. Three preferred "topologies" were selected from several hundred thousand possible topologies of these seven predicted strands using a rule-based analysis. The subsequently determined experimental structure showed that these seven strands in synaptotagmin adopt one of the three preferred topologies. We were unable, however, to identify the correct topology from among these three topologies. PMID- 8710838 TI - Ab initio structure prediction for small polypeptides and protein fragments using genetic algorithms. AB - Ab initio folding simulations have been performed on three peptides, using a genetic algorithm-based search method which operates on a full atom representation. Conformations are evaluated with an empirical force field parameterized by a potential of mean force analysis of experimental structures. The dominant terms in the force field are local and nonlocal main chain electrostatics and the hydrophobic effect. Two of the simulated structures were for fragments of complete proteins (eosinophil-derived neurotoxin (EDN) and the subtilisin propeptide) that were identified as being likely initiation sites for folding. The experimental structure of one of these (EDN) was subsequently found to be consistent with that prediction (using local hydrophobic burial as the determinant for independent folding). The simulations of the structures of these two peptides were only partly successful. The most successful folding simulation was that of a 22-residue peptide corresponding to the membrane binding domain of blood coagulation factor VIII (Membind). Three simulations were performed on this peptide and the lowest energy conformation was found to be the most similar to the experimental structure. The conformation of this peptide was determined with a C alpha rms deviation of 4.4 A. Although these simulations were partly successful there are still many unresolved problems, which we expect to be able to address in the next structure prediction experiment. PMID- 8710839 TI - Bacterial toxins deliver the goods. PMID- 8710840 TI - RNA editing in plant organelles: a fertile field. PMID- 8710841 TI - Ras-catalyzed hydrolysis of GTP: a new perspective from model studies. AB - Despite the biological and medical importance of signal transduction via Ras proteins and despite considerable kinetic and structural studies of wild-type and mutant Ras proteins, the mechanism of Ras-catalyzed GTP hydrolysis remains controversial. We take a different approach to this problem: the uncatalyzed hydrolysis of GTP is analyzed, and the understanding derived is applied to the Ras-catalyzed reaction. Evaluation of previous mechanistic proposals from this chemical perspective suggests that proton abstraction from the attacking water by a general base and stabilization of charge development on the gamma-phosphoryl oxygen atoms would not be catalytic. Rather, this analysis focuses attention on the GDP leaving group, including the beta-gamma bridge oxygen of GTP, the atom that undergoes the largest change in charge in going from the ground state to the transition state. This leads to a new catalytic proposal in which a hydrogen bond from the backbone amide of Gly-13 to this bridge oxygen is strengthened in the transition state relative to the ground state, within an active site that provides a template complementary to the transition state. Strengthened transition state interactions of the active site lysine, Lys-16, with the beta nonbridging phosphoryl oxygens and a network of interactions that positions the nucleophilic water molecule and gamma-phosphoryl group with respect to one another may also contribute to catalysis. It is speculated that a significant fraction of the GAP-activated GTPase activity of Ras arises from an additional interaction of the beta-gamma bridge oxygen with an Arg side chain that is provided in trans by GAP. The conclusions for Ras and related G proteins are expected to apply more widely to other enzymes that catalyze phosphoryl (-PO(3)2 ) transfer, including kinases and phosphatases. PMID- 8710842 TI - Meiosis: how could it work? AB - A physical connection between homologs is required for reductional segregation at the first division of meiosis. This connection is usually provided by one or a few well-spaced crossovers. A speculative overview of processes leading to formation of these crossovers is presented. PMID- 8710844 TI - A joint hazard and time scaling model to compare survival curves. AB - To provide a more general method for comparing survival experience, we propose a model that independently scales both hazard and time dimensions. To test the curve shape similarity of two time-dependent hazards, h1(t) and h2(t), we apply the proposed hazard relationship, h12(tKt)/ h1(t) = Kh, to h1. This relationship doubly scales h1 by the constant hazard and time scale factors, Kh and Kt, producing a transformed hazard, h12, with the same underlying curve shape as h1. We optimize the match of h12 to h2 by adjusting Kh and Kt. The corresponding survival relationship S12(tKt) = [S1(t)]KtKh transforms S1 into a new curve S12 of the same underlying shape that can be matched to the original S2. We apply this model to the curves for regional and local breast cancer contained in the National Cancer Institute's End Results Registry (1950-1973). Scaling the original regional curves, h1 and S1 with Kt = 1.769 and Kh = 0.263 produces transformed curves h12 and S12 that display congruence with the respective local curves, h2 and S2. This similarity of curve shapes suggests the application of the more complete curve shapes for regional disease as templates to predict the long-term survival pattern for local disease. By extension, this similarity raises the possibility of scaling early data for clinical trial curves according to templates of registry or previous trial curves, projecting long-term outcomes and reducing costs. The proposed model includes as special cases the widely used proportional hazards (Kt = 1) and accelerated life (KtKh = 1) models. PMID- 8710843 TI - Molecular control of vertebrate iron metabolism: mRNA-based regulatory circuits operated by iron, nitric oxide, and oxidative stress. AB - As an essential nutrient and a potential toxin, iron poses an exquisite regulatory problem in biology and medicine. At the cellular level, the basic molecular framework for the regulation of iron uptake, storage, and utilization has been defined. Two cytoplasmic RNA-binding proteins, iron-regulatory protein-1 (IRP-1) and IRP-2, respond to changes in cellular iron availability and coordinate the expression of mRNAs that harbor IRP-binding sites, iron-responsive elements (IREs). Nitric oxide (NO) and oxidative stress in the form of H2O2 also signal to IRPs and thereby influence cellular iron metabolism. The recent discovery of two IRE-regulated mRNAs encoding enzymes of the mitochondrial citric acid cycle may represent the beginnings of elucidating regulatory coupling between iron and energy metabolism. In addition to providing insights into the regulation of iron metabolism and its connections with other cellular pathways, the IRE/IRP system has emerged as a prime example for the understanding of translational regulation and mRNA stability control. Finally, IRP-1 has highlighted an unexpected role for iron sulfur clusters as post-translational regulatory switches. PMID- 8710845 TI - A designed beta-hairpin peptide in crystals. AB - Beta-hairpin structures have been crystallographically characterized only in very short acyclic peptides, in contrast to helices. The structure of the designed beta-hairpin, t-butoxycarbonyl-Leu-Val-Val-D-Pro-Gly-Leu-Val-Val-OMe in crystals is described. The two independent molecules of the octapeptide fold into almost ideal beta-hairpin conformations with the central D-Pro-Gly segment adopting a Type II' beta-turn conformation. The definitive characterization of a beta hairpin has implications for de novo peptide and protein design, particularly for the development of three- and four-stranded beta-sheets. PMID- 8710846 TI - Enzyme-mediated spatial segregation on individual polymeric support beads: application to generation and screening of encoded combinatorial libraries. AB - Proteolysis of short N alpha-protected peptide substrates bound to polyoxyethylene-polystyrene beads releases selectively free amino sites in the enzyme-accessible "surface" area. The substantial majority of functional sites in the "interior" of the polymeric support are not reached by the enzyme and remain uncleaved (protected). Subsequent synthesis with two classes of orthogonal protecting groups-N alpha-tert-butyloxycarbonyl (Boc) and N alpha-9 fluorenylmethyloxy-carbonyl (Fmoc)-allows generation of two structures on the same bead. The surface structure is available for receptor interactions, whereas the corresponding interior structure is used for coding. Coding structures are usually readily sequenceable peptides. This "shaving" methodology was illustrated by the preparation of a peptide-encoded model peptide combinatorial library containing 1.0 x 10(5) members at approximately 6-fold degeneracy. From this single library, good ligands were selected for three different receptors: anti beta-endorphin anti-body, streptavidin, and thrombin, and the binding structures were deduced correctly by sequencing the coding peptides present on the same beads. PMID- 8710847 TI - Photochemical energy conversion in a helical oligoproline assembly. AB - A general method is described for constructing a helical oligoproline assembly having a spatially ordered array of functional sites protruding from a proline-II helix. Three different redox-active carboxylic acids were coupled to the side chain of cis-4-amino-L-proline. These redox modules were incorporated through solid-phase peptide synthesis into a 13-residue helical oligoproline assembly bearing in linear array a phenothiazine electron donor, a tris(bipyridine)ruthenium(II) chromophore, and an anthraquinone electron acceptor. Upon transient 460-nm irradiation in acetonitrile, this peptide triad formed with 53% efficiency an excited state containing a phenothiazine radical cation and an anthraquinone radical anion. This light-induced redox-separated state had a lifetime of 175 ns and stored 1.65 eV of energy. PMID- 8710848 TI - Location of a potential transport binding site in a sigma class glutathione transferase by x-ray crystallography. AB - The crystal structure of the sigma class glutathione transferase from squid digestive gland in complex with S-(3-iodobenzyl)glutathione reveals a third binding site for the glutathione conjugate besides the two in the active sites of the dimer. The additional binding site is near the crystallographic two-fold axis between the two alpha 4-turn-alpha 5 motifs. The principal binding interactions with the conjugate include specific electrostatic interactions between the peptide and the two subunits and a hydrophobic cavity found across the two-fold axis that accommodates the 3-iodobenzyl group. Thus, two identical, symmetry related but mutually exclusive binding modes for the third conjugate are observed. The hydrophobic pocket is about 14 A from the hydroxyl group of Tyr-7 in the active site. This site is a potential transport binding site for hydrophobic molecules or their glutathione conjugates. PMID- 8710849 TI - Targeted disruption of the arylsulfatase B gene results in mice resembling the phenotype of mucopolysaccharidosis VI. AB - Mucopolysaccharidosis VI (MPS VI) is a lysosomal storage disease with autosomal recessive inheritance caused by a deficiency of the enzyme arylsulfatase B (ASB), which is involved in degradation of dermatan sulfate and chondroitin 4-sulfate. A MPS VI mouse model was generated by targeted disruption of the ASB gene. Homozygous mutant animals exhibit ASB enzyme deficiency and elevated urinary secretion of dermatan sulfate. They develop progressive symptoms resembling those of MPS VI in humans. Around 4 weeks of age facial dysmorphia becomes overt, long bones are shortened, and pelvic and costal abnormalities are observed. Major alterations in bone formation with perturbed cartilaginous tissues in newborns and widened, perturbed, and persisting growth plates in adult animals are seen. All major parenchymal organs show storage of glycosaminoglycans preferentially in interstitial cells and macrophages. Affected mice are fertile and mortality is not elevated up to 15 months of age. This mouse model will be a valuable tool for studying pathogenesis of MPS VI and may help to evaluate therapeutical approaches for lysosomal storage diseases. PMID- 8710850 TI - NMR evidence for the participation of a low-barrier hydrogen bond in the mechanism of delta 5-3-ketosteroid isomerase. AB - Delta 5-3-Ketosteroid isomerase (EC 5.3.3.1) promotes an allylic rearrangement involving intramolecular proton transfer via a dienolic intermediate. This enzyme enhances the catalytic rate by a factor of 10(10). Two residues, Tyr-14, the general acid that polarizes the steroid 3-carbonyl group and facilitates enolization, and Asp-38 the general base that abstracts and transfers the 4 beta proton to the 6 beta-position, contribute 10(4.7) and 10(5.6) to the rate increase, respectively. A major mechanistic enigma is the huge disparity between the pKa values of the catalytic groups and their targets. Upon binding of an analog of the dienolate intermediate to isomerase, proton NMR detects a highly deshielded resonance at 18.15 ppm in proximity to aromatic protons, and with a 3 fold preference for protium over deuterium (fractionation factor, phi = 0.34), consistent with formation of a short, strong (low-barrier) hydrogen bond to Tyr 14. The strength of this hydrogen bond is estimated to be at least 7.1 kcal/mol. This bond is relatively inaccessible to bulk solvent and is pH insensitive. Low barrier hydrogen bonding of Tyr-14 to the intermediate, in conjunction with the previously demonstrated tunneling contribution to the proton transfer by Asp-38, provide a plausible and quantitative explanation for the high catalytic power of this isomerase. PMID- 8710851 TI - Engineering actin-resistant human DNase I for treatment of cystic fibrosis. AB - Human deoxyribonuclease I (DNase I), an enzyme recently approved for treatment of cystic fibrosis (CF), has been engineered to create two classes of mutants: actin resistant variants, which still catalyze DNA hydrolysis but are no longer inhibited by globular actin (G-actin) and active site variants, which no longer catalyze DNA hydrolysis but still bind G-actin. Actin-resistant variants with the least affinity for actin, as measured by an actin binding ELISA and actin inhibition of [33P] DNA hydrolysis, resulted from the introduction of charged, aliphatic, or aromatic residues at Ala-114 or charged residues on the central hydrophobic actin binding interface at Tyr-65 or Val-67. In CF sputum, the actin resistant variants D53R, Y65A, Y65R, or V67K were 10-to 50-fold more potent than wild type in reducing viscoelasticity as determined in sputum compaction assays. The reduced viscoelasticity correlated with reduced DNA length as measured by pulsed-field gel electrophoresis. In contrast, the active site variants H252A or H134A had no effect on altering either viscoelasticity or DNA length in CF sputum. The data from both the active site and actin-resistant variants demonstrate that the reduction of viscoelasticity by DNase I results from DNA hydrolysis and not from depolymerization of filamentous actin (F-actin). The increased potency of the actin-resistant variants indicates that G-actin is a significant inhibitor of DNase I in CF sputum. These results further suggest that actin-resistant DNase I variants may have improved efficacy in CF patients. PMID- 8710852 TI - The primary structures of the Archaeon Halobacterium salinarium blue light receptor sensory rhodopsin II and its transducer, a methyl-accepting protein. AB - Recently, a large family of transducer proteins in the Archaeon Halobacterium salinarium was identified. On the basis of the comparison of the predicted structural domains of these transducers, three distinct subfamilies of transducers were proposed. Here we report isolation, complete gene sequences, and analysis of the encoded primary structures of transducer gene htrII, a member of family B, and its blue light receptor gene (sopII) of sensory rhodopsin II (SRII). The start codon ATG of the 714-bp sopII gene is one nucleotide beyond the termination codon TGA of the 2298-bp htrII gene. The deduced protein sequence of HtrII predicts a eubacterial chemotaxis transducer type with two hydrophobic membrane-spanning segments connecting sizable domains in the periplasm and cytoplasm. HtrII has a common feature with HtrI, the sensory rhodopsin I transducer; like HtrI, HtrII possesses a hydrophilic loop structure just after the second transmembrane segment. The C-terminal 299 residues (765 amino acid residues total) of HtrII show strong homology to the signaling and methylation domain of eubacterial transducer Tsr. The hydropathy plot of the primary structure of SRII indicates seven membrane-spanning alpha-helical segments, a characteristic feature of retinylidene proteins ("rhodopsins") from a widespread family of photoactive pigments. SRII shows high identity with SRI (42%), bacteriorhodopsin (BR) (32%), and halorhodopsin (24%). The crucial positions for retinal binding sites in these proteins are nearly identical, with the exception of Met-118 (numbering according to the mature BR sequence), which is replaced by Val in SRII. In BR, residues Asp-85 and Asp-96 are crucial in proton pumping. In SRII, the position corresponding to Asp-85 in BR is conserved, but the corresponding position of Asp-96 is replaced by an aromatic Tyr. Coexpression of the htrII and sopII genes restores SRII phototaxis to a mutant (Pho81) that contains a deletion in the htrI/sopI and insertion in htrII/sopII regions. This paper describes the first example that both HtrI and HtrII exist in the same halobacterial cell, confirming that different sensory rhodopsins SRI and SRII in the same organism have their own distinct transducers. PMID- 8710854 TI - I-TRAF is a novel TRAF-interacting protein that regulates TRAF-mediated signal transduction. AB - Tumor necrosis factor (TNF) receptor-associated factor (TRAF) proteins associate with and transduce signals from TNF receptor 2, CD40, and presumably other members of the TNF receptor superfamily. TRAF2 is required for CD40- and TNF mediated activation of the transcription factor NF-kappa B. Here we describe the isolation and characterization of a novel TRAF-interacting protein, I-TRAF, that binds to the conserved TRAF-C domain of the three known TRAFs. Overexpression of I-TRAF inhibits TRAF2-mediated NF-kappa B activation signaled by CD40 and both TNF receptors. Thus, I-TRAF appears as a natural regulator of TRAF function that may act by maintaining TRAFs in a latent state. PMID- 8710853 TI - Mammalian Sug1 and c-Fos in the nuclear 26S proteasome. AB - In a search for regulatory proteins that interact with the leucine zipper motif of c-Fos in the yeast two-hybrid screen, we have identified a protein (FZA-B) that has extensive sequence similarity to SUG1 of Saccharomyces cerevisiae. Here we show that FZA-B can functionally substitute for SUG1 in yeast and that FZA-B interacts with Fos proteins in vitro through their leucine zippers. In rat liver and in HeLa cells, FZA-B is present in the 26S proteasome complex, as is c-Fos. Immobilized antibody raised against an FZA-B-specific peptide depleted peptidase activity, proteasomal proteins, FZA-B, and c-Fos from a 26S proteasome preparation. FZA-B is found predominantly in the nuclear fraction of COS cells expressing an FZA-B transgene and in the nuclear 26S proteasome of HeLa cells. We conclude that FZA-B is the mammalian homolog of SUG1 (mSug1) and that it is present in the nuclear 26S proteasome of cells. Our results suggest that mSug1 may be involved in the degradation of c-Fos and other transcription factors. PMID- 8710855 TI - Sequence and expression of the murine Hoxd-3 homeobox gene. AB - Murine Hoxd-3 (Hox 4.1) genomic DNA and cDNA and Hoxa-3 (Hox 1.5) cDNA were cloned and sequenced. The homeodomains of Hoxd-3 and Hoxa-3 and regions before and after the homeodomain are highly conserved. Both Hoxa-3 and Hoxa-3 proteins have a proline-rich region that contains consensus amino acid sequences for binding to Src homology 3 domains of some signal transduction proteins. Northern blot analysis of RNA from 8- to 11-day-old mouse embryos revealed a 4.3-kb species of Hoxd-3 RNA, whereas a less abundant 3.0-kb species of Hoxd-3 RNA was found in RNA from 9- to 11-day-old embryos. Two species of Hoxd-3 poly(A)+ RNA, 4.3 and 6.0 kb in length, were found in poly(A)+ RNA from adult mouse kidney, but not in RNA from other adult tissues tested. Hoxd-3 mRNA was detected by in situ hybridization in 12-, 14-, and 17-day-old mouse embryos in the posterior half of the myelencephalon, spinal cord, dorsal root ganglia, first cervical vertebra, thyroid gland, kidney tubules, esophagus, stomach, and intestines. PMID- 8710856 TI - A hyperphosphorylated form of the large subunit of RNA polymerase II is associated with splicing complexes and the nuclear matrix. AB - A hyperphosphorylated form of the largest subunit of RNA polymerase II (pol IIo) is associated with the pre-mRNA splicing process. Pol IIo was detected in association with a subset of small nuclear ribonucleoprotein particle and Ser-Arg protein splicing factors and also with pre-mRNA splicing complexes assembled in vitro. A subpopulation of pol IIo was localized to nuclear "speckle" domains enriched in splicing factors, indicating that it may also be associated with RNA processing in vivo. Moreover, pol IIo was retained in a similar pattern following in situ extraction of cells and was quantitatively recovered in the nuclear matrix fraction. The results implicate nuclear matrix-associated hyperphosphorylated pol IIo as a possible link in the coordination of transcription and splicing processes. PMID- 8710857 TI - Transformation of Rat-1 fibroblasts with the v-src oncogene increases the tyrosine phosphorylation state and activity of the alpha subunit of Gq/G11. AB - Two major intermediaries in signal transduction pathways are pp60v-sre family tyrosine kinases and heterotrimeric guanine nucleotide-binding proteins. In Rat-1 fibroblasts transformed by the v-src oncogene, endothelin-1 (ET-1)-induced inositol 1,4,5-trisphosphate accumulation is increased 6-fold, without any increases in the numbers of ET-1 receptors or in the response to another agonist, thrombin. This ET-1 hyperresponse can be inhibited by an antibody directed against the carboxyl terminus of the Gq/G11 alpha subunit, suggesting that the Gq/G11 protein couples ET-1 receptors to phospholipase C (PLC). While v-src transformation did not increase the expression of the Gq/G11 alpha subunit, immunoblotting with anti-phosphotyrosine antibodies and phosphoamino acid analysis demonstrated that the Gq/G11 alpha subunit becomes phosphorylated on tyrosine residues in v-src-transformed cells. Moreover, when the Gq/G11 protein was extracted from control and transformed cell lines and reconstituted with exogenous PLC, AIF*4-stimulated Gq/G11 activity was markedly increased in extracts from v-src-transformed cells. Our results demonstrate that the process of v-src transformation can increase the tyrosine phosphorylation state of the Gq/G11 alpha-subunit in intact cells and that the process causes an increase in the Gq/G11 alpha-subunit's ability to stimulate PLC following activation with AIF 4. PMID- 8710858 TI - Sequence tag identification of intact proteins by matching tanden mass spectral data against sequence data bases. AB - Molecular and fragment ion data of intact 8- to 43-kDa proteins from electrospray Fourier-transform tandem mass spectrometry are matched against the corresponding data in sequence data bases. Extending the sequence tag concept of Mann and Wilm for matching peptides, a partial amino acid sequence in the unknown is first identified from the mass differences of a series of fragment ions, and the mass position of this sequence is defined from molecular weight and the fragment ion masses. For three studied proteins, a single sequence tag retrieved only the correct protein from the data base; a fourth protein required the input of two sequence tags. However, three of the data base proteins differed by having an extra methionine or by missing an acetyl or heme substitution. The positions of these modifications in the protein examined were greatly restricted by the mass differences of its molecular and fragment ions versus those of the data base. To characterize the primary structure of an unknown represented in the data base, this method is fast and specific and does not require prior enzymatic or chemical degradation. PMID- 8710859 TI - Migration of vesicular stomatitis virus glycoprotein to the nucleus of infected cells. AB - A new means of direct visualization of the early events of viral infection by selective fluorescence labeling of viral proteins coupled with digital imaging microscopy is reported. The early phases of viral infection have great importance for understanding viral replication and pathogenesis. Vesicular stomatitis virus, the best-studied rhabdovirus, is composed of an RNA genome of negative sense, five viral proteins, and membrane lipids derived from the host cell. The glycoprotein of vesicular stomatitis virus was labeled with fluorescein isothiocyanate, and the labeled virus was incubated with baby hamster kidney cells. After initiation of infection, the fluorescence of the labeled glycoprotein was first seen inside the cells in endocytic vesicles. The fluorescence progressively migrated to the nucleus of infected cells. After 1 h of infection, the virus glycoprotein was concentrated in the nucleus and could be recovered intact in a preparation of purified nuclei. These results suggest that uncoating of the viral RNA occurs close to the nuclear membrane, which would precede transcription of the leader RNA that enters the nucleus to shut off cellular RNA synthesis and DNA replication. PMID- 8710860 TI - Association of yeast SIN1 with the tetratrico peptide repeats of CDC23. AB - The yeast SIN1 protein is a nuclear protein that together with other proteins behaves as a transcriptional repressor of a family of genes. In addition, sin1 mutants are defective in proper mitotic chromosome segregation. In an effort to understand the basis for these phenotypes, we employed the yeast two-hybrid system to identify proteins that interact with SIN1 in vivo. Here we demonstrate that CDC23, a protein known to be involved in sister chromatid separation during mitosis, is able to directly interact with SIN1. Furthermore, using recombinant molecules in vitro, we show that the N terminal of SIN1 is sufficient to bind a portion of CDC23 consisting solely of tetratrico peptide repeats. Earlier experiments identified the C-terminal domain of SIN1 to be responsible for interaction with a protein that binds the regulatory region of HO, a gene whose transcription is repressed by SIN1. Taken together with the results presented here, we suggest that SIN1 is a chromatin protein having at least a dual function: The N terminal of SIN1 interacts with the tetratrico peptide repeat domains of CDC23, a protein involved in chromosome segregation, whereas the C terminal of SIN1 binds proteins involved in transcriptional regulation. PMID- 8710861 TI - p34cdc2 kinase activity is maintained upon activation of the replication checkpoint in Schizosaccharomyces pombe. AB - All eukaryotes use feedback controls to order and coordinate cell cycle events. In Schizosaccharomyces pombe, several classes of checkpoint genes serve to ensure that DNA replication is complete and free of error before the onset of mitosis. Wild-type cells normally arrest upon inhibition of DNA synthesis or in response to DNA damage, although the exact mechanisms controlling this arrest are unclear. Genetic evidence in fission yeast suggests that the dependence of mitosis upon completion of DNA replication is linked to the regulation of the p34cdc2 cyclin dependent kinase. It has been hypothesized that inhibition of DNA synthesis triggers down-regulation of p34cdc2 kinase activity, although this has never been shown biochemically. We analyzed the activity of p34cdc2 in wild-type and checkpoint-defective cells treated with a DNA synthesis inhibitor. Using standard in vitro assays we demonstrate that p34cdc2 kinase activity is maintained in wild type cells arrested at the replication checkpoint. We also used a novel in vivo assay for p34cdc2 kinase activity, in which we expressed a fragment of the human retinoblastoma tumor suppressor protein in fission yeast. Phosphorylation of this fragment of the human retinoblastoma tumor suppressor protein is dependent on p34cdc2 kinase activity, and this activity is also maintained in cells arrested at the replication checkpoint. These data suggest that the mechanism for cell cycle arrest in response to incomplete DNA synthesis is not dependent on the attenuation of p34cdc2 activity. PMID- 8710862 TI - Reactivation of denatured proteins by 23S ribosomal RNA: role of domain V. AB - Escherichia coli ribosome, its 50S subunit, or simply the 23S rRNA can reactivate denatured proteins in vitro. Here we show that protein synthesis inhibitors chloramphenicol and erythromycin, which bind to domain V of 23S rRNA of E. coli, can inhibit reactivation of denatured pig muscle lactate dehydrogenase and fungal glucose-6-phosphate dehydrogenase by 23S rRNA completely. Oligodeoxynucleotides complementary to two regions within domain V (which cover sites of chloramphenicol resistant mutations and the putative A site of the incoming aminoacyl tRNA), but not to a region outside of domain V, also can inhibit the activity. Domain V of 23S rRNA, therefore, appears to play a crucial role in reactivation of denatured proteins. PMID- 8710863 TI - Active heterodimers are formed from human DNA topoisomerase II alpha and II beta isoforms. AB - DNA topoisomerase II is a nuclear enzyme essential for chromosome dynamics and DNA metabolism. In mammalian cells, two genetically and biochemically distinct topoisomerase II forms exist, which are designated topoisomerase II alpha and topoisomerase II beta. In our studies of human topoisomerase II, we have found that a substantial fraction of the enzyme exists as alpha/beta heterodimers in HeLa cells. The ability to form heterodimers was verified when human topoisomerases II alpha and II beta were coexpressed in yeast and investigated in a dimerization assay. Analysis of purified heterodimers shows that these enzymes maintain topoisomerase II specific catalytic activities. The natural existence of an active heterodimeric subclass of topoisomerase II merits attention whenever topoisomerases II alpha and II beta function, localization, and cell cycle regulation are investigated. PMID- 8710864 TI - Dual signal transduction through delta opioid receptors in a transfected human T cell line. AB - Opiates are known to function as immunomodulators, in part by effects on T cells. However, the signal transduction pathways mediating the effects of opiates on T cells are largely undefined. To determine whether pathways that regulate free intracellular calcium ([Ca2+]i) and/or cAMP are affected by opiates acting through delta-type opioid receptors (DORs), a cDNA encoding the neuronal DOR was expressed in a stably transfected Jurkat T-cell line. The DOR agonists, deltorphin and [D-Ala2, D-Leu5]-enkephalin (DADLE), elevated [Ca2+]i, measured by flow cytofluorometry using the calcium-sensitive dye, Fluo-3. At concentrations from 10(-11)-10(-7) M, both agonists increased [Ca2+]i from 60 nM to peak concentrations of 400 nM in a dose-dependent manner within 30 sec (ED50 of approximately 5 x 10(-9) M). Naltrindole, a selective DOR antagonist, abolished the increase in [Ca2+]i, and pretreatment with pertussis toxin was also effective. To assess the role of extracellular calcium, cells were pretreated with EGTA, which reduced the initial deltorphin-induced elevation of [Ca2+]i by more than 50% and eliminated the second phase of calcium mobilization. Additionally, the effect of DADLE on forskolin-stimulated cAMP production was determined. DADLE reduced cAMP production by 70% (IC50 of approximately equal to 10(-11) M), and pertussis toxin inhibited the action of DADLE. Thus, the DOR expressed by a transfected Jurkat T-cell line is positively coupled to pathways leading to calcium mobilization and negatively coupled to adenylate cyclase. These studies identify two pertussis toxin-sensitive, G protein-mediated signaling pathways through which DOR agonists regulate the levels of intracellular messengers that modulate T-cell activation. PMID- 8710865 TI - Evidence for aminoacylation-induced conformational changes in human mitochondrial tRNAs. AB - Analysis by acid polyacrylamide/urea gel electrophoresis of 14 individual mitochondrial tRNAs (mt-tRNAs) from human cells has revealed a variable decrease in mobility of the aminoacylated relative to the nonacylated form, with the degree of separation of the two forms not being correlated with the mass, polar character, or charge of the amino acid. Separation of the charged and uncharged species has been found to be independent of tRNA denaturation, being observed also in the absence of urea. In another approach, electrophoresis through a perpendicular denaturing gradient gel of several individual mt-tRNAs has shown a progressive unfolding of the tRNA with increasing denaturant concentration, which is consistent with an initial disruption of tertiary interactions, followed by the sequential melting of the four stems of the cloverleaf structure. A detailed analysis of the unfolding process of charged and uncharged tRNALys and tRNALeu(UUR) has revealed that the separation of the two forms of these tRNAs persisted throughout the almost entire range of denaturant concentrations used and was lost upon denaturation of the last helical domain(s), which most likely included the amino acid acceptor stem. These observations strongly suggest that the electrophoretic retardation of the charged species reflects an aminoacylation induced conformational change of the 3'-end of these mt-tRNAs, with possible significant implications in connection with the known role of the acceptor end in tRNA interactions with the ribosomal peptidyl transferase center and the elongation factor Tu. PMID- 8710866 TI - Binding of a hairpin polyamide in the minor groove of DNA: sequence-specific enthalpic discrimination. AB - Hairpin polyamides are synthetic ligands for sequence-specific recognition in the minor groove of double-helical DNA. A thermodynamic characterization of the DNA binding properties exhibited by a six-ring hairpin polyamide, ImPyPy-gamma-PyPyPy beta-Dp (where Im = imidazole, Py = pyrrole, gamma = gamma-aminobutyric acid, beta = beta-alanine, and Dp = dimethylaminopropylamide), reveals an approximately 1-2 kcal/mol greater affinity for the designated match site, 5'-TGTTA-3', relative to the single base pair mismatch sites, 5'-TGGTA-3' and 5'-TATTA-3'. The enthalpy and entropy data at 20 degrees C reveal this sequence specificity to be entirely enthalpic in origin. Correlations between the thermodynamic driving forces underlying the sequence specificity exhibited by ImPyPy-gamma-PyPyPy-beta Dp and the structural properties of the heterodimeric complex of PyPyPy and ImPyPy bound to the minor groove of DNA provide insight into the molecular forces that govern the affinity and specificity of pyrrole-imidazole polyamides. PMID- 8710867 TI - The solution structure of the Raf-1 cysteine-rich domain: a novel ras and phospholipid binding site. AB - The Raf-1 protein kinase is the best-characterized downstream effector of activated Ras. Interaction with Ras leads to Raf-1 activation and results in transduction of cell growth and differentiation signals. The details of Raf-1 activation are unclear, but our characterization of a second Ras-binding site in the cysteine-rich domain (CRD) and the involvement of both Ras-binding sites in effective Raf-1-mediated transformation provides insight into the molecular aspects and consequences of Ras-Raf interactions. The Raf-1 CRD is a member of an emerging family of domains, many of which are found within signal transducing proteins. Several contain binding sites for diacylglycerol (or phorbol esters) and phosphatidylserine and are believed to play a role in membrane translocation and enzyme activation. The CRD from Raf-1 does not bind diacylglycerol but interacts with Ras and phosphatidylserine. To investigate the ligand-binding specificities associated with CRDs, we have determined the solution structure of the Raf-1 CRD using heteronuclear multidimensional NMR. We show that there are differences between this structure and the structures of two related domains from protein kinase C (PKC). The differences are confined to regions of the CRDs involved in binding phorbol ester in the PKC domains. Since phosphatidylserine is a common ligand, we expect its binding site to be located in regions where the structures of the Raf-1 and PKC domains are similar. The structure of the Raf-1 CRD represents an example of this family of domains that does not bind diacylglycerol and provides a framework for investigating its interactions with other molecules. PMID- 8710868 TI - Wild-type and mutant p53 differentially regulate transcription of the insulin like growth factor I receptor gene. AB - The insulin-like growth factor I receptor (IGF-I-R) plays a critical role in transformation events. It is highly overexpressed in most malignant tissues where it functions as an anti-apoptotic agent by enhancing cell survival. Tumor suppressor p53 is a nuclear transcription factor that blocks cell cycle progression and induces apoptosis. p53 is the most frequently mutated gene in human cancer. Cotransfection of Saos-2 (os-teosarcoma-derived cells) and RD (rhabdomyosarcoma-derived cells) cells with IGF-I-R promoter constructs driving luciferase reporter genes and with wild-type p53 expression vectors suppressed promoter activity in a dose-dependent manner. This effect of p53 is mediated at the level of transcription and it involves interaction with TBP, the TATA box binding component of TFIID. On the other hand, three tumor-derived mutant forms of p53 (mut 143, mut 248, and mut 273) stimulated the activity of the IGF-I-R promoter and increased the levels of IGF-I-R/luciferase fusion mRNA. These results suggest that wild-type p53 has the potential to suppress the IGF-I-R promoter in the postmitotic, fully differentiated cell, thus resulting in low levels of receptor gene expression in adult tissues. Mutant versions of p53 protein, usually associated with malignant states, can derepress the IGF-I-R promoter, with ensuing mitogenic activation by locally produced or circulating IGFs. PMID- 8710869 TI - Multiple tyrosine residues in the cytosolic domain of the erythropoietin receptor promote activation of STAT5. AB - Signaling through the erythropoietin receptor (EPO-R) is crucial for proliferation, differentiation, and survival of erythroid progenitor cells. EPO induces homodimerization of the EPO-R, triggering activation of the receptor associated kinase JAK2 and activation of STAT5. By mutating the eight tyrosine residues in the cytosolic domain of the EPO-R, we show that either Y343 or Y401 is sufficient to mediate maximal activation of STAT5; tyrosine residues Y429 and Y431 can partially activate STAT5. Comparison of the sequences surrounding these tyrosines reveals YXXL as the probable motif specifying recruitment of STAT5 to the EPO-R. Expression of a mutant EPO-R lacking all eight tyrosine residues in the cytosolic domain supported a low but detectable level of EPO-induced STAT5 activation, indicating the existence of an alternative pathway for STAT5 activation independent of any tyrosine in the EPO-R. The kinetics of STAT5 activation and inactivation were the same, regardless of which tyrosine residue in the EPO-R mediated its activation or whether the alternative pathway was used. The ability of mutant EPO-Rs to activate STAT5 did not directly correlate with their mitogenic potential. PMID- 8710870 TI - Ligand induction of a transcriptionally active thyroid hormone receptor coactivator complex. AB - Transcriptional regulation by nuclear hormone receptors is thought to involve interactions with putative cofactors that may potentiate receptor function. Here we show that human thyroid hormone receptor alpha purified from HeLa cells grown in the presence of thyroid hormone (T3) is associated with a group of distinct nuclear proteins termed thyroid hormone receptor-associated proteins (TRAPs). In an in vitro system reconstituted with general initiation factors and cofactors (and in the absence of added T3), the "liganded" thyroid hormone receptor (TR)/TRAP complex markedly activates transcription from a promoter template containing T3-response elements. Moreover, whereas the retinoid X receptor is not detected in the TR/TRAP complex, its presence is required for the function of the complex. In contrast, human thyroid hormone receptor alpha purified from cells grown in the absence of T3 lacks the TRAPs and effects only a low level of activation that is dependent on added ligand. These findings demonstrate the ligand-dependent in vivo formation of a transcriptionally active TR-multisubunit protein complex and suggest a role for TRAPs as positive coactivators for gene specific transcriptional activation. PMID- 8710871 TI - Methylation of coding region alone inhibits gene expression in plant protoplasts. AB - Derivatives of the cauliflower mosaic virus 35S promoter lacking CG and CNG methylation targets were constructed and used to direct transcription of reporter gene constructs in transiently transformed protoplasts. Such methylation-target free (MTF) promoters, although weaker than the 35S promoter, retain significant activity despite mutation of the as-1 element. The effect of methylation on gene expression in MTF- and 35S-promoter driven constructs was examined. Even when the promoter region was free of methylation targets, reporter gene expression was markedly reduced when cytosine residues in CG dinucleotides were methylated in vitro prior to transformation. Mosaic methylation experiments, in which only specific parts of the plasmids were methylated, revealed that methylation of the coding region alone has a negative effect on reporter gene expression. Methylation nearer the 5' end of the coding region was more inhibitory, consistent with inhibition of transcription elongation. PMID- 8710872 TI - Do cyanobacteria swim using traveling surface waves? AB - Bacteria that swim without the benefit of flagella might do so by generating longitudinal or transverse surface waves. For example, swimming speeds of order 25 microns/s are expected for a spherical cell propagating longitudinal waves of 0.2 micron length, 0.02 micron amplitude, and 160 microns/s speed. This problem was solved earlier by mathematicians who were interested in the locomotion of ciliates and who considered the undulations of the envelope swept out by ciliary tips. A new solution is given for spheres propagating sinusoidal waveforms rather than Legendre polynomials. The earlier work is reviewed and possible experimental tests are suggested. PMID- 8710873 TI - Characterizations of diverse residue clusters in protein three-dimensional structures. AB - We present new methods for identifying and analyzing statistically significant residue clusters that occur in three-dimensional (3D) protein structures. Residue clusters of different kinds occur in many contexts. They often feature the active site (e.g., in substrate binding), the interface between polypeptide units of protein complexes, regions of protein-protein and protein-nucleic acid interactions, or regions of metal ion coordination. The methods are illustrated with 3D clusters centering on four themes. (i) Acidic or histidine-acidic clusters associated with metal ions. (ii) Cysteine clusters including coordination of metals such as zinc or iron-sulfur structures, cysteine knots prominent in growth factors, multiple sets of buried disulfide pairings that putatively nucleate the hydrophobic core, or cysteine clusters of mostly exposed disulfide bridges. (iii) Iron-sulfur proteins and charge clusters. (iv) 3D environments of multiple histidine residues. Study of diverse 3D residue clusters offers a new perspective on protein structure and function. The algorithms can aid in rapid identification of distinctive sites, suggest correlations among protein structures, and serve as a tool in the analysis of new structures. PMID- 8710874 TI - Clusters of charged residues in protein three-dimensional structures. AB - Statistically significant charge clusters (basic, acidic, or of mixed charge) in tertiary protein structures are identified by new methods from a large representative collection of protein structures. About 10% of protein structures show at least one charge cluster, mostly of mixed type involving about equally anionic and cationic residues. Positive charge clusters are very rare. Negative (or histidine-acidic) charge clusters often coordinate calcium, or magnesium or zinc ions [e.g., thermolysin (PDB code: 3tln), mannose-binding protein (2msb), aminopeptidase (1amp)]. Mixed-charge clusters are prominent at interchain contacts where they stabilize quaternary protein formation [e.g., glutathione S transferase (2gst), catalase (8act), and fructose-1,6-bisphosphate aldolase (1fba)]. They are also involved in protein-protein interaction and in substrate binding. For example, the mixed-charge cluster of aspartate carbamoyl-transferase (8atc) envelops the aspartate carbonyl substrate in a flexible manner (alternating tense and relaxed states) where charge associations can vary from weak to strong. Other proteins with charge clusters include the P450 cytochrome family (BM-3, Terp, Cam), several flavocytochromes, neuraminidase, hemagglutinin, the photosynthetic reaction center, and annexin. In each case in Table 2 we discuss the possible role of the charge clusters with respect to protein structure and function. PMID- 8710875 TI - The folding mechanism of larger model proteins: role of native structure. AB - The folding mechanism of a 125-bead heteropolymer model for proteins is investigated with Monte Carlo simulations on a cubic lattice. Sequences that do and do not fold in a reasonable time are compared. The overall folding behavior is found to be more complex than that of models for smaller proteins. Folding begins with a rapid collapse followed by a slow search through the semi-compact globule for a sequence-dependent stable core with about 30 out of 176 native contacts which serves as the transition state for folding to a near-native structure. Efficient search for the core is dependent on structural features of the native state. Sequences that fold have large amounts of stable, cooperative structure that is accessible through short-range initiation sites, such as those in anti-parallel sheets connected by turns. Before folding is completed, the system can encounter a second bottleneck, involving the condensation and rearrangement of surface residues. Overly stable local structure of the surface residues slows this stage of the folding process. The relation of the results from the 125-mer model studies to the folding of real proteins is discussed. PMID- 8710876 TI - Ultra-fast excited state dynamics in green fluorescent protein: multiple states and proton transfer. AB - The green fluorescent protein (GFP) of the jellyfish Aequorea Victoria has attracted widespread interest since the discovery that its chromophore is generated by the autocatalytic, posttranslational cyclization and oxidation of a hexapeptide unit. This permits fusion of the DNA sequence of GFP with that of any protein whose expression or transport can then be readily monitored by sensitive fluorescence methods without the need to add exogenous fluorescent dyes. The excited state dynamics of GFP were studied following photo-excitation of each of its two strong absorption bands in the visible using fluorescence upconversion spectroscopy (about 100 fs time resolution). It is shown that excitation of the higher energy feature leads very rapidly to a form of the lower energy species, and that the excited state interconversion rate can be markedly slowed by replacing exchangeable protons with deuterons. This observation and others lead to a model in which the two visible absorption bands correspond to GFP in two ground-state conformations. These conformations can be slowly interconverted in the ground state, but the process is much faster in the excited state. The observed isotope effect suggests that the initial excited state process involves a proton transfer reaction that is followed by additional structural changes. These observations may help to rationalize and motivate mutations that alter the absorption properties and improve the photo stability of GFP. PMID- 8710877 TI - Ultraviolet-induced movement of the human DNA repair protein, Xeroderma pigmentosum type G, in the nucleus. AB - Xeroderma pigmentosum type G (XPG) is a human genetic disease exhibiting extreme sensitivity to sunlight. XPG patients are defective XPG endonuclease, which is an enzyme essential for DNA repair of the major kinds of solar ultraviolet (UV) induced DNA damages. Here we describe a novel dynamics of this protein within the cell nucleus after UV irradiation of human cells. Using confocal microscopy, we have localized the immunofluorescent, antigenic signal of XPG protein to foci throughout the cell nucleus. Our biochemical studies also established that XPG protein forms a tight association with nuclear structure(s). In human skin fibroblast cells, the number of XPG foci decreased within 2 h after UV irradiation, whereas total nuclear XPG fluorescence intensity remained constant, suggesting redistribution of XPG from a limited number of nuclear foci to the nucleus overall. Within 8 h after UV, most XPG antigenic signal was found as foci. Using beta-galactosidase-XPG fusion constructs (beta-gal-XPG) transfected into HeLa cells, we have identified a single region of XPG that is evidently responsible both for foci formation and for the UV dynamic response. The fusion protein carrying the C terminus of XPG (amino acids 1146-1185) localized beta-gal specific antigenic signal to foci and to the nucleolus regions. After UV irradiation, antigenic beta-gal translocated reversibly from the subnuclear structures to the whole nucleus with kinetics very similar to the movements of XPG protein. These findings lead us to propose a model in which distribution of XPG protein may regulate the rate of DNA repair within transcriptionally active and inactive compartments of the cell nucleus. PMID- 8710878 TI - The full-length leptin receptor has signaling capabilities of interleukin 6-type cytokine receptors. AB - The leptin receptor (OB-R) is a single membrane-spanning protein that mediates the weight regulatory effects of leptin (OB protein). The mutant allele (db) of the OB-R gene encodes a protein with a truncated cytoplasmic domain that is predicted to be functionally inactive. Several mRNA splice variants encoding OB Rs with different length cytoplasmic domains have been detected in various tissues. Here we demonstrate that the full-length OB-R (predominantly expressed in the hypothalamus), but not a major naturally occurring truncated form or a mutant from found in db/db mice, can mediate activation of signal transducer and activator of transcription (STAT) proteins and stimulate transcription through interleukin 6 responsive gene elements. Reconstitution experiments suggest that, although OB-R mediates intracellular signals with a specificity similar to interleukin 6-type cytokine receptors, signaling appears to be independent of the gp130 signal transducing component of the interleukin 6-type cytokine receptors. PMID- 8710879 TI - Mutant conformation of p53 translated in vitro or in vivo requires functional HSP90. AB - The p53 mutant, 143ala, was translated in vitro in either rabbit reticulocyte lysate (RRL) or wheat germ extract (WGE). In RRL, p53-143ala protein of both mutant and wild-type conformation, as detected immunologically with conformation specific antibodies, was translated. The chaperone protein HSP90, present in RRL, was found to coprecipitate only with the mutated conformation of p53. Geldanamycin, shown previously to bind to HSP90 and destabilize its association with other proteins, decreased the amount of immunologically detectable mutated p53 and increased the amount of detectable wild-type protein, without affecting the total translation of p53. When translated in WGE, known to contain functionally deficient HSP90, p53-143ala produced p53 protein, which was not recognized by a mutated conformation-specific antibody. In contrast, the synthesis of conformationally detectable wild-type p53 in this system was not compromised. Reconstitution of HSP90 function in WGE permitted synthesis of conformationally detectable mutated p53, and this was abrogated by geldanamycin. Finally, when p53-143ala was stably tansfected into yeast engineered to be defective for HSP90 function, conformational recognition of mutated p53 was impaired. When stable transfectants of p53-143ala were prepared in yeast expressing wild-type HSP90, conformational recognition of mutated p53 was antagonized by macbecin I, a geldanamycin analog also known to bind HSP90. Taken together, these data demonstrate a role for HSP90 in the achievement and/or stabilization of the mutated conformation of p53-143ala. Furthermore, we show that the mutated conformation of p53 can be pharmacologically antagonized by drugs targeting HSP90. PMID- 8710880 TI - Specific accumulation of tumor-derived adhesion factor in tumor blood vessels and in capillary tube-like structures of cultured vascular endothelial cells. AB - Tumor-derived adhesion factor (TAF) was previously identified as a cell adhesion molecule secreted by human bladder carcinoma cell line EJ-1. To elucidate the physiological function of TAF, we examined its distribution in human normal and tumor tissues. Immunochemical staining with an anti-TAF monoclonal antibody showed that TAF was specifically accumulated in small blood vessels and capillaries within and adjacent to tumor nests, but not in those in normal tissues. Tumor blood vessel-specific staining of TAF was observed in various human cancers, such as esophagus, brain, lung, and stomach cancers. Double immunofluorescent staining showed apparent colocalization of TAF and type IV collagen in the vascular basement membrane. In vitro experiments demonstrated that TAF preferentially bound to type IV collagen among various extracellular matrix components tested. In cell culture experiments, TAF promoted adhesion of human umbilical vein endothelial cells to type IV collagen substrate and induced their morphological change. Furthermore, when the endothelial cells were induced to form capillary tube-like structures by type I collagen, TAF and type IV collagen were exclusively detected on the tubular structures. The capillary tube formation in vitro was prevented by heparin, which inhibited the binding of TAF to the endothelial cells. These results strongly suggest that TAF contributes to the organization of new capillary vessels in tumor tissues by modulating the interaction of endothelial cells with type IV collagen. PMID- 8710881 TI - Mammalian cells resistant to tumor suppressor genes. AB - Expression of p53 causes growth arrest or apoptosis in many normal and neoplastic cell types, but the relationship between these two effects has remained obscure. To begin to dissect the underlying mechanisms at a genetic level, we have generated mutant cells resistant to the action of wild-type p53. Rat embryo fibroblasts transformed with ras and a temperature-sensitive p53 (tsp53(135val)) gene were chemically mutagenized and selected for growth at a temperature at which p53 adopts a wild-type conformation (31.5 degrees C). Clones that grew exponentially at 31.5 degrees C were selected. Cell fusion experiments demonstrated that the mutations conferring resistance to p53-mediated growth arrest were dominant. The mutagenized clones were resistant not only to p53 mediated growth arrest, but also to the apoptosis induced by E1A in conjunction with p53, and partially resistant to the retinoblastoma tumor suppressor, pRB. The results suggest that a single downstream pathway can control the induction of growth arrest and apoptosis, and that both p53 and RB function through this pathway. PMID- 8710882 TI - Cleavage of lamin A by Mch2 alpha but not CPP32: multiple interleukin 1 beta converting enzyme-related proteases with distinct substrate recognition properties are active in apoptosis. AB - Although proteases related to the interleukin 1 beta-converting enzyme (ICE) are known to be essential for apoptotic execution, the number of enzymes involved, their substrate specificities, and their specific roles in the characteristic biochemical and morphological changes of apoptosis are currently unknown. These questions were addressed using cloned recombinant ICE-related proteases (IRPs) and a cell-free model system for apoptosis (S/M extracts). First, we compared the substrate specificities of two recombinant human IRPs, CPP32 and Mch2 alpha. Both enzymes cleaved poly-(ADP-ribose) polymerase, albeit with different efficiencies. Mch2 alpha also cleaved recombinant and nuclear lamin A at a conserved VEID decreases NG sequence located in the middle of the coiled-coil rod domain, producing a fragment that was indistinguishable from the lamin A fragment observed in S/M extracts and in apoptotic cells. In contrast, CPP32 did not cleave lamin A. The cleavage of lamin A by Mch2 alpha and by S/M extracts was inhibited by millimolar concentrations of Zn2+, which had a minimal effect on cleavage of poly (ADP-ribose) polymerase by CPP32 and by S/M extracts. We also found that N-(acetyltyrosinylvalinyl-N epsilon-biotinyllysyl)aspartic acid [(2,6 dimethylbenzoyl)oxy]methyl ketone, which derivatizes the larger subunit of active ICE, can affinity label up to five active IRPs in S/M extracts. Together, these observations indicate that the processing of nuclear proteins in apoptosis involves multiple IRPs having distinct preferences for their apoptosis-associated substrates. PMID- 8710883 TI - Evidence for an insulin receptor substrate 1 independent insulin signaling pathway that mediates insulin-responsive glucose transporter (GLUT4) translocation. AB - Interaction of the activated insulin receptor (IR) with its substrate, insulin receptor substrate 1 (IRS-1), via the phosphotyrosine binding domain of IRS-1 and the NPXY motif centered at phosphotyrosine 960 of the IR, is important for IRS-1 phosphorylation. We investigated the role of this interaction in the insulin signaling pathway that stimulates glucose transport. Utilizing microinjection of competitive inhibitory reagents in 3T3-L1 adipocytes, we have found that disruption of the IR/IRS-1 interaction has no effect upon translocation of the insulin-responsive glucose transporter (GLUT4). The activity of these reagents was demonstrated by their ability to block insulin stimulation of two distinct insulin bioeffects, membrane ruffling and mitogenesis, in 3T3-L1 adipocytes and insulin-responsive rat 1 fibroblasts. These data suggest that phosphorylated IRS 1 is not an essential component of the metabolic insulin signaling pathway that leads to GLUT4 translocation, yet it appears to be required for other insulin bioeffects. PMID- 8710884 TI - Microspectroscopic imaging tracks the intracellular processing of a signal transduction protein: fluorescent-labeled protein kinase C beta I. AB - We have devised a microspectroscopic strategy for assessing the intracellular (re)distribution and the integrity of the primary structure of proteins involved in signal transduction. The purified proteins are fluorescent-labeled in vitro and reintroduced into the living cell. The localization and molecular state of fluorescent-labeled protein kinase C beta I isozyme were assessed by a combination of quantitative confocal laser scanning microscopy, fluorescence lifetime imaging microscopy, and novel determinations of fluorescence resonance energy transfer based on photobleaching digital imaging microscopy. The intensity and fluorescence resonance energy transfer efficiency images demonstrate the rapid nuclear translocation and ensuing fragmentation of protein kinase C beta I in BALB/c3T3 fibroblasts upon phorbol ester stimulation, and suggest distinct, compartmentalized roles for the regulatory and catalytic fragments. PMID- 8710885 TI - Toxoplasma invasion: the parasitophorous vacuole is formed from host cell plasma membrane and pinches off via a fission pore. AB - Most intracellular pathogens avoid lysing their host cells during invasion by wrapping themselves in a vacuolar membrane. This parasitophorous vacuole membrane (PVM) is often retained, serving as a critical transport interface between the parasite and the host cell cytoplasm. To test whether the PVM formed by the parasite Toxoplasma gondii is derived from host cell membrane or from lipids secreted by the parasite, we used time-resolved capacitance measurements and video microscopy to assay host cell surface area during invasion. We observed no significant change in host cell surface area during PVM formation, demonstrating that the PVM consists primarily of invaginated host cell membrane. Pinching off of the PVM from the host cell membrane occurred after an unexpected delay (34-305 sec) and was seen as a 0.219 +/- 0.006 pF drop in capacitance, which corresponds well to the predicted surface area of the entire PVM (30-33 microns2). The formation and closure of a fission pore connecting the extracellular medium and the vacuolar space was detected as the PVM pinched off. This final stage of parasite entry was accomplished without any breach in cell membrane integrity. PMID- 8710886 TI - Muc1, a mucin-like protein that is regulated by Mss10, is critical for pseudohyphal differentiation in yeast. AB - Pseudohyphal differentiation in Saccharomyces cerevisiae was first described as a response of diploid cells to nitrogen limitation. Here we report that haploid and diploid starch-degrading S. cerevisiae strains were able to switch from a yeast form to a filamentous pseudohyphal form in response to carbon limitation in the presence of an ample supply of nitrogen. Two genes, MSS10 and MUC1, were cloned and shown to be involved in pseudohyphal differentiation and invasive growth. The deletion of MSS10 resulted in extremely reduced amounts of pseudohyphal differentiation and invasive growth, whereas the deletion of MUC1 abolished pseudohyphal differentiation and invasive growth completely. Mss10 appears to be a transcriptional activator that responds to nutrient limitation and coregulates the expression of MUC1 and the STA1-3 glucoamylase genes, which are involved in starch degradation. MUC1 encodes a 1367-amino acid protein, containing several serine/threonine-rich repeats. Muc1 is a putative integral membrane-bound protein, similar to mammalian mucin-like membrane proteins that have been implicated to play a role in the ability of cancer cells to invade other tissues. PMID- 8710887 TI - Cellular adherence elicits ligand-independent activation of the Met cell-surface receptor. AB - Cell adhesion has a fundamental role in the proliferation and motility of normal cells and the metastasis of tumor cells. To identify signaling pathways activated by the adherence of tumor cells, we analyzed the tyrosine phosphorylation of proteins in mouse melanoma cells before and after attachment to substrata. We discovered that cellular adherence activated the protein-tyrosine kinase of the cell surface receptor Met, whose ligand is hepatocyte growth factor and scatter factor. The activation was exceedingly prompt, affected the great majority of Met in the cells, persisted so long as the cells remained adherent, and was rapidly reversed as soon as the cells were detached from substrata. Activation of Met required that cells be adherent but not that they spread on the substratum, and it occurred in the absence of any apparent ligand for the receptor. Ligand independent activation of Met occurred in several varieties of tumor cells but not in normal endothelial cells that express the receptor. The activation of Met described here may represent a means by which cells respond to mechanical as opposed to biochemical stimuli. PMID- 8710888 TI - Antibody-induced engagement of beta 2 integrins on adherent human neutrophils triggers activation of p21ras through tyrosine phosphorylation of the protooncogene product Vav. AB - It is known that beta 2 integrins are crucial for leukocyte cell-cell and cell matrix interactions, and accumulating evidence now suggests that integrins serve not only as a structural link but also as a signal-transducing unit that controls adhesion-induced changes in cell functions. In the present study, we plated human neutrophils on surface-bound anti-beta 2 (CD18) antibodies and found that the small GTP-binding protein p21ras is activated by beta 2 integrins. Pretreatment of the cells with genistein, a tyrosine kinase inhibitor, led to a complete block of p21ras activation, an effect that was not achieved with either U73122, which abolishes the beta 2 integrin-induced Ca2+ signal, or wortmannin, which totally inhibits the phosphatidylinositol 3-kinase activity. Western blot analysis revealed that antibody-induced engagement of beta 2 integrins causes tyrosine phosphorylation of several proteins in the cells. One of these tyrosine phosphorylated proteins had an apparent molecular mass of 95 kDa and was identified as the protooncogene product Vav, a p21ras guanine nucleotide exchange factor that is specifically expressed in cells of hematopoietic lineage. A role for Vav in the activation of p21ras is supported by the observations that antibody-induced engagement of beta 2 integrins causes an association of Vav with p21ras and that the effect of genistein on p21ras activation coincided with its ability to inhibit both the tyrosine phosphorylation of Vav and the Vav-p21ras association. Taken together, these results indicate that antibody-induced engagement of beta 2 integrins on neutrophils triggers tyrosine phosphorylation of Vav and, possibly through its association, a downstream activation of p21ras. PMID- 8710889 TI - Fused polycationic peptide mediates delivery of diphtheria toxin A chain to the cytosol in the presence of anthrax protective antigen. AB - The lethal factor (LF) and edema factor (EF) of anthrax toxin bind by means of their amino-terminal domains to protective antigen (PA) on the surface of toxin sensitive cells and are translocated to the cytosol, where they act on intracellular targets. Genetically fusing the amino-terminal domain of LF (LFN; residues 1-255) to certain heterologous proteins has been shown to potentiate these proteins for PA-dependent delivery to the cytosol. We report here that short tracts of lysine, arginine, or histidine residues can also potentiate a protein for such PA-dependent delivery. Fusion of these polycationic tracts to the amino terminus of the enzymic A chain of diphtheria toxin (DTA; residues 1 193) enabled it to be translocated to the cytosol by PA and inhibit protein synthesis. The efficiency of translocation was dependent on tract length: (LFN > Lys8 > Lys6 > Lys3). Lys6 was approximately 100-fold more active than Arg6 or His6, whereas Glu6 and (SerSerGly)2 were inactive. Arg6DTA was partially degraded in cell culture, which may explain its low activity relative to that of Lys6DTA. The polycationic tracts may bind to anionic sites at the cell surface (possibly on PA), allowing the fusion proteins to be coendocytosed with PA and delivered to the endosome, where translocation to the cytosol occurs. Excess free LFN blocked the action of LFNDTA, but not of Lys6DTA. This implies that binding to the LF/EF site is not an obligatory step in translocation and suggests that the polycationic tag binds to a different site. Besides elucidating the process of translocation in anthrax toxin, these findings may aid in developing systems to deliver heterologous proteins and peptides to the cytoplasm of mammalian cells. PMID- 8710890 TI - Cloning and characterization of KAP3: a novel kinesin superfamily-associated protein of KIF3A/3B. AB - We previously reported that KIF3A and KIF3B form a heterodimer that functions as a microtubule-based fast anterograde translocator of membranous organelles. We have also shown that this KIF3A/3B forms a complex with other associated polypeptides, named kinesin superfamily-associated protein 3 (KAP3). In the present study, we purified KAP3 protein by immunoprecipitation using anti-KIF3B antibody from mouse testis. Microsequencing was carried out, and we cloned the full-length KAP3 cDNA from a mouse brain cDNA library. Two isoforms of KAP3 exist [KAP3A (793 aa) and KAP3B (772 aa)], generated by alternative splicing in the carboxyl terminus region. Their amino acid sequences have no homology with those of any other known proteins, and prediction of their secondary structure indicated that almost the entire KAP3 molecule is alpha-helical. We produced recombinant KAP3 and KIF3A/3B using a baculovirus-Sf9 expression system. A reconstruction study in Sf9 cells revealed that KAP3 is a globular protein that binds to the tail domain of KIF3A/3B. The immunolocalization pattern of KAP3 was similar to that of KIF3A/3B in nerve cells. In addition, we found that KAP3 does not affect the motor activity of KIF3A/3B. KAP3 was associated with a membrane bound form of KIF3A/3B in a fractional immunoprecipitation experiment, and since the KIF3 complex was found to bind to membranous organelles in an EM study, KAP3 may regulate membrane binding of the KIF3 complex. PMID- 8710891 TI - Reduced DNA methylation in Arabidopsis thaliana results in abnormal plant development. AB - Arabidopsis plants transformed with an antisense construct of an Arabidopsis methyltransferase cDNA (METI) have reduced cytosine methylation in CG dinucleotides. Methylation levels in progeny of five independent transformants ranged from 10% to 100% of the wild type. Removal of the antisense construct by segregation in sexual crosses did not fully restore methylation patterns in the progeny, indicating that methylation patterns are subject to meiotic inheritance in Arabidopsis. Plants with decreased methylation displayed a number of phenotypic and developmental abnormalities, including reduced apical dominance, smaller plant size, altered leaf size and shape, decreased fertility, and altered flowering time. Floral organs showed homeotic transformations that were associated with ectopic expression of the floral homeotic genes AGAMOUS and APETALA3 in leaf tissue. These observations suggest that DNA methylation plays an important role in regulating many developmental pathways in plants and that the developmental abnormalities seen in the methyltransferase antisense plants may be due to dysregulation of gene expression. PMID- 8710892 TI - A molecular mechanism for the effect of lithium on development. AB - Lithium, one of the most effective drugs for the treatment of bipolar (manic depressive) disorder, also has dramatic effects on morphogenesis in the early development of numerous organisms. How lithium exerts these diverse effects is unclear, but the favored hypothesis is that lithium acts through inhibition of inositol monophosphatase (IMPase). We show here that complete inhibition of IMPase has no effect on the morphogenesis of Xenopus embryos and present a different hypothesis to explain the broad action of lithium. Our results suggest that lithium acts through inhibition of glycogen synthase kinase-3 beta (GSK-3 beta), which regulates cell fate determination in diverse organisms including Dictyostelium, Drosophila, and Xenopus. Lithium potently inhibits GSK-3 beta activity (Ki = 2 mM), but is not a general inhibitor of other protein kinases. In support of this hypothesis, lithium treatment phenocopies loss of GSK-3 beta function in Xenopus and Dictyostelium. These observations help explain the effect of lithium on cell-fate determination and could provide insights into the pathogenesis and treatment of bipolar disorder. PMID- 8710893 TI - Defective forebrain development in mice lacking gp330/megalin. AB - gp330/megalin, a member of the low density lipoprotein (LDL) receptor gene family, is expressed on the apical surfaces of epithelial tissues, including the neuroepithelium, where it mediates the endocytic uptake of diverse macromolecules, such as cholesterol-carrying lipoproteins, proteases, and antiproteinases. Megalin knockout mice manifest abnormalities in epithelial tissues including lung and kidney that normally express the protein and they die perinatally from respiratory insufficiency. In brain, impaired proliferation of neuroepithelium produces a holoprosencephalic syndrome, characterized by lack of olfactory bulbs, forebrain fusion, and a common ventricular system. Similar syndromes in humans and animals are caused by insufficient supply of cholesterol during development. Because megalin can bind lipoproteins, we propose that the receptor is part of the maternal-fetal lipoprotein transport system and mediates the endocytic uptake of essential nutrients in the postgastrulation stage. PMID- 8710894 TI - The notion of the Cambrian pananimalia genome. AB - The toil by photosynthesizing cyanobacteria and blue-green algae of nearly three billion years appeared to have finally resulted in the sufficient accumulation of molecular oxygen. So, the stage was set for the emergence, at the ocean bottom, of diverse animals that were consumers of molecular oxygen. It now appears that this Cambrian explosion, during which nearly all the extant animal phyla have emerged, was of an astonishingly short duration, lasting only 6-10 million years. Inasmuch as only a 1% DNA base sequence change is expected in 10 million years under the standard spontaneous mutation rate, I propose that all those diverse animals of the early Cambrian period, some 550 million years ago, were endowed with nearly identical genomes, with differential usage of the same set of genes accounting for the extreme diversities of body forms. Some of the more pertinent genes that are thought to be included in the Cambrian pananimalia genome are as follows. (i) A gene for lysyloxidase that, in the presence of molecular oxygen, crosslinked collagen triple helices to produce ligaments and tendons, thus contributing to the stout bodies of the Cambrian animals. (ii) Genes for hemoglobin; these internal transporters of molecular oxygen are today seen sporadically in members of diverse animal phyla. (iii) The Pax-6 gene for eye formation; the eyes of a ribbon worm to a human are organized by this gene. In animals without eyes, the same gene organizes other sensory systems and organs. (iv) A series of Hox genes for the anterior-posterior (cranio-caudal) body plans: these genes are also present in all phyla of the kingdom Animalia. PMID- 8710895 TI - Class 3 Hox genes in insects and the origin of zen. AB - We have cloned, from a beetle and a locust, genes that are homologous to the class 3 Hox genes of vertebrates. Outside the homeobox they share sequence motifs with the Drosophila zerknullt (zen) and z2 genes, and like zen, are expressed only in extraembryonic membranes. We conclude that the zen genes of Drosophila derive from a Hox class 3 sequence that formed part of the common ancestral Hox cluster, but that in insects this (Hox) gene has lost its role in patterning the anterio-posterior axis of the embryo, and acquired a new function. In the lineage leading to Drosophila, the zen genes have diverged particularly rapidly. PMID- 8710896 TI - Translocation events in the evolution of aminoacyl-tRNA synthetases. AB - We have characterized hisS, the gene encoding the histidyl-tRNA synthetase (HisRS) from the tetraodontoid fish Fugu rubripes. The hisS gene is about 3.5 kbp long and contains 13 exons and 12 introns of 172 bp, on average. The Fugu hisS gene encodes a putative protein of 519 amino acids with the three motifs identified as signatures of class 2 aminoacyl-tRNA synthetases. A model for the shifting of intron 8 between Fugu and hamster is proposed based on the successive appearance of a cryptic splicing site followed by an insertion mutation that created a new acceptor site. In addition, sequence comparisons suggest that the hisS gene has undergone a translocation through the first intron. As a result, the Fugu HisRS has an N-terminal sequence markedly different from that in the human and hamster enzymes. We propose that similar events have been responsible for variations at the N-terminal end of other aminoacyl-tRNA synthetases. Our analysis suggests that this involves exchanges through introns of two exons encoding an ancestral 32-amino acid motif. PMID- 8710897 TI - Identification of interdomain sequences promoting the intronless evolution of a bacterial protein family. AB - In the evolution of eukaryotic genes, introns are believed to have played a major role in increasing the probability of favorable duplication events, chance recombinations, and exon shuffling resulting in functional hybrid proteins. As a rule, prokaryotic genes lack introns, and the examples of prokaryotic introns described do not seem to have contributed to gene evolution by exon shuffling. Still, certain protein families in modern bacteria evolve rapidly by recombination of genes, duplication of functional domains, and as shown for protein PAB of the anaerobic bacterial species Peptostreptococcus magnus, by the shuffling of an albumin-binding protein module from group C and G streptococci. Characterization of a protein PAB-related gene in a P. magnus strain with less albumin-binding activity revealed that the shuffled module was missing. Based on this fact and observations made when comparing gene sequences of this family of bacterial surface proteins interacting with albumin and/or immunoglobulin, a model is presented that can explain how this rapid intronless evolution takes place. A new kind of genetic element is introduced: the recer sequence promoting interdomain, in frame recombination and acting as a structure-less flexibility promoting spacer in the corresponding protein. The data presented also suggest that antibiotics could represent the selective pressure behind the shuffling of protein modules in P. magnus, a member of the indigenous bacterial flora. PMID- 8710898 TI - A three-hybrid system to detect RNA-protein interactions in vivo. AB - RNA-protein interactions are pivotal in fundamental cellular processes such as translation, mRNA processing, early development, and infection by RNA viruses. However, in spite of the central importance of these interactions, few approaches are available to analyze them rapidly in vivo. We describe a yeast genetic method to detect and analyze RNA-protein interactions in which the binding of a bifunctional RNA to each of two hybrid proteins activates transcription of a reporter gene in vivo. We demonstrate that this three-hybrid system enables the rapid, phenotypic detection of specific RNA-protein interactions. As examples, we use the binding of the iron regulatory protein 1 (IRP1) to the iron response element (IRE), and of HIV trans-activator protein (Tat) to the HIV trans activation response element (TAR) RNA sequence. The three-hybrid assay we describe relies only on the physical properties of the RNA and protein, and not on their natural biological activities; as a result, it may have broad application in the identification of RNA-binding proteins and RNAs, as well as in the detailed analysis of their interactions. PMID- 8710899 TI - Suppression of glioblastoma angiogenicity and tumorigenicity by inhibition of endogenous expression of vascular endothelial growth factor. AB - The development of new capillary networks from the normal microvasculature of the host appears to be required for growth of solid tumors. Tumor cells influence this process by producing both inhibitors and positive effectors of angiogenesis. Among the latter, the vascular endothelial growth factor (VEGF) has assumed prime candidacy as a major positive physiological effector. Here, we have directly tested this hypothesis in the brain tumor, glioblastoma multiforme, one of the most highly vascularized human cancers. We introduced an antisense VEGF expression construct into glioblastoma cells and found that (i) VEGF mRNA and protein levels were markedly reduced, (ii) the modified cells did not secrete sufficient factors so as to be chemoattractive for primary human microvascular endothelial cells, (iii) the modified cells were not able to sustain tumor growth in immunodeficient animals, and (iv) the density of in vivo blood vessel formation was reduced in direct relation to the reduction of VEGF secretion and tumor formation. Moreover, revertant cells that recovered the ability to secrete VEGF regained each of these tumorigenic properties. These results suggest that VEGF plays a major angiogenic role in glioblastoma. PMID- 8710900 TI - Simultaneous fluorescence-activated cell sorter analysis of two distinct transcriptional elements within a single cell using engineered green fluorescent proteins. AB - Green fluorescent protein (GFP) is widely used as a reporter gene in both prokaryotes and eukaryotes. However, the fluorescence levels of wild-type GFP (wtGFP) are not bright enough for fluorescence-activated cell sorting or flow cytometry. Several GFP variants were generated that are brighter or have altered excitation spectra when expressed in prokaryotic cells. We engineered two GFP genes with different combinations of these mutations, GFP(S65T,V163A) termed GFP Bex1, and GFP(S202F,T203I,V163A) termed GFP-Vex1. Both show enhanced brightness and improved signal-to-noise ratios when expressed in mammalian cells and appropriately excited, compared with wtGFP. Each mutant retains only one of the two excitation peaks of the wild-type protein. GFP-Bex1 excites at 488 nm (blue) and GFP-Vex1 excites at 406 nm (violet), both of which are available laser lines. Excitation at these wavelengths allows for the independent analyses of these mutants by fluorescence-activated cell sorting, permitting simultaneous, quantitative detection of expression from two different genes within single mammalian cells. PMID- 8710901 TI - Induction of centromeric activity in maize by suppressor of meiotic drive 1. AB - The Abnormal chromosome 10 (Ab10) in maize causes normally-quiescent blocks of heterochromatin called knobs to function as meiotic centromeres. Under these circumstances genetic markers associated with knobs exhibit meiotic drive, i.e., they are preferentially transmitted to progeny. Here we describe a mutation called suppressor of meiotic drive (smd1) that partially suppresses meiotic drive, and demonstrate that smd1 causes a quantitative reduction in the mobility of knobs on the meiotic spindle. We conclude that Smd1 encodes a product that is necessary for the activation of ectopic centromeres, and that meiotic drive occurs as a consequence of the resulting change in chromosome movement. As a genetic system, Ab10 offers a new and powerful approach for analyzing centromere/kinetochore function. PMID- 8710902 TI - Antisense ribosomes: rRNA as a vehicle for antisense RNAs. AB - Although rRNA has a conserved core structure, its size varies by more than 2000 bases between eubacteria and vertebrates, mostly due to the size variation of discrete variable regions. Previous studies have shown that insertion of foreign sequences into some of these variable regions has little effect on rRNA function. These properties make rRNA a potentially very advantageous vehicle to carry other RNA moieties with biological activity, such as "antisense RNAs." We have explored this possibility by inserting antisense RNAs targeted against one essential and two nonessential genes into a site within a variable region in the Tetrahymena thermophila large subunit rRNA gene. Expression of each of the three genes tested can be drastically reduced or eliminated in transformed T. thermophila lines containing these altered rRNAs. In addition, we found that only antisense rRNAs containing RNA sequences complementary to the 5' untranslated region of the targeted mRNA were effective. Lines containing antisense rRNAs targeted against either of the nonessential genes grow well, indicating that the altered rRNAs fulfill their functions within the ribosome. Since functional rRNA is extremely abundant and stable and comes into direct contact with translated mRNAs, it may prove to be an unparalleled vehicle for enhancing the activity of functional RNAs that act on mRNAs. PMID- 8710903 TI - A computer-based systematic survey reveals the predominance of small inverted repeat elements in wild-type rice genes. AB - Several recent reports indicate that mobile elements are frequently found in and flanking many wild-type plant genes. To determine the extent of this association, we performed computer-based systematic searches to identify mobile elements in the genes of two "model" plants, Oryza sativa (domesticated rice) and Arabidopsis thaliana. Whereas 32 common sequences belonging to nine putative mobile element families were found in the noncoding regions of rice genes, none were found in Arabidopsis genes. Five of the nine families (Gaijin, Castaway, Ditto, Wanderer, and Explorer) are first described in this report, while the other four were described previously (Tourist, Stowaway, p-SINE1, and Amy/LTP). Sequence similarity, structural similarity, and documentation of past mobility strongly suggests that many of the rice common sequences are bona fide mobile elements. Members of four of the new rice mobile element families are similar in some respects to members of the previously identified inverted-repeat element families, Tourist and Stowaway. Together these elements are the most prevalent type of transposons found in the rice genes surveyed and form a unique collection of inverted-repeat transposons we refer to as miniature inverted-repeat transposable elements or MITEs. The sequence and structure of MITEs are clearly distinct from short or long interspersed nuclear elements (SINEs or LINEs), the most common transposable elements associated with mammalian nuclear genes. Mobile elements, therefore, are associated with both animal and plant genes, but the identity of these elements is strikingly different. PMID- 8710904 TI - Constitutive overexpression of Cu/Zn superoxide dismutase exacerbates kainic acid induced apoptosis of transgenic-Cu/Zn superoxide dismutase neurons. AB - Cu/Zn superoxide dismutase (Cu/Zn SOD) is a key enzyme in the metabolism of oxygen free radicals. The gene resides on chromosome 21 and is overexpressed in patients with Down syndrome. Cultured neurons of transgenic Cu/Zn SOD (Tg-Cu/Zn SOD) mice with elevated activity of Cu/Zn SOD were used to determine whether constitutive overexpression of Cu/Zn SOD creates an indigenous oxidative stress that predisposes the Tg-Cu/Zn SOD neurons to added insults. Neurons from three independently derived Tg-Cu/Zn SOD strains showed higher susceptibility than nontransgenic neurons to kainic acid (KA)-mediated excitotoxicity, reflected by an earlier onset and enhanced apoptotic cell death. This higher susceptibility of transgenic neurons to KA-mediated apoptosis was associated with a chronic prooxidant state that was manifested by reduced levels of cellular glutathione and altered [Ca2+]i homeostasis. The data are compatible with the thesis that overexpression of Cu/Zn SOD creates chronic oxidative stress in the transgenic neurons, which exacerbates their susceptibility to additional insults such as KA mediated excitotoxicity. PMID- 8710905 TI - Sin3 corepressor function in Myc-induced transcription and transformation. AB - Many basic-helix-loop-helix-leucine zipper (b-HLH-LZ) proteins, including the Myc family and non-Myc family, bind a common DNA sequence CACGTG, yet have quite different biological actions. Myc binds this sequence as a heterodimer with Max in the activation of both transcription and transformation. The Myc family members Mad and Mxi1 are known to suppress Myc-induced transcription and transformation and to dimerize with Max to form ternary complexes with the mammalian Sin3 transcriptional corepressor (mSin3). The b-HLH-LZ domain of TFEB, which cannot heterodimerize within the Myc family, does not suppress Myc-induced transcription or transformation. However, transfer of a 25- to 36-aa region from Mad or Mxi1, which interacts with mSin3, to the b-HLH-LZ of TFEB, mediated profound suppression of Myc-induced transcription and transformation. These results suggest that the DNA binding specificities of the Myc family and non-Myc family b-HLH-LZ proteins, in the context of the cellular genes involved in Myc induced transformation, are shared. The results also demonstrate that targeting mSin3 to CACGTG sites via a non-Myc family DNA binding domain is sufficient to oppose Myc activity in growth regulation. PMID- 8710907 TI - Multiple forms of complement C3 in trout that differ in binding to complement activators. AB - In all other species analyzed to date, the functionally active form of complement component C3 exists as the product of a single gene. We have now identified and characterized three functional C3 proteins (C3-1, C3-3, and C3-4) in trout that are the products of at least two distinct C3 genes. All three proteins are composed of an alpha-and a beta-chain and contain a thioester bond in the alpha chain. However, they differ in their electrophoretic mobility, glycosylation, reactivity with monospecific C3 antibodies, and relative ability to bind to various surfaces (zymosan, Escherichia coli, erythrocytes). A comparison of the partial amino acid sequences of the three proteins showed that the amino acid sequence identity/similarity of C3-3 to C3-4 is 87/91%, while that of C3-3 and C3 4 to C3-1 is 51.5/65.5% and 60/73% respectively. Thus, trout possess multiple forms of functional C3 that represent the products of several distinct genes and differ in their ability to bind covalently to various complement activators. PMID- 8710906 TI - Prevalence of germ-line mutations in p16, p19ARF, and CDK4 in familial melanoma: analysis of a clinic-based population. AB - Five to ten percent of individuals with melanoma have another affected family member, suggesting familial predisposition. Germ-line mutations in the cyclin dependent kinase (CDK) inhibitor p16 have been reported in a subset of melanoma pedigrees, but their prevalence is unknown in more common cases of familial melanoma that do not involve large families with multiple affected members. We screened for germ-line mutations in p16 and in two other candidate melanoma genes, p19ARF and CDK4, in 33 consecutive patients treated for melanoma; these patients had at least one affected first or second degree relative (28 independent families). Five independent, definitive p16 mutations were detected (18%, 95% confidence interval: 6%, 37%), including one nonsense, one disease associated missense, and three small deletions. No mutations were detected in CDK4. Disease-associated mutations in p19ARF, whose transcript is derived in part from an alternative codon reading frame of p16, were only detected in patients who also had mutations inactivating p16. We conclude that germ-line p16 mutations are present in a significant fraction of individuals who have melanoma and a positive family history. PMID- 8710908 TI - Identification of the zinc-dependent endothelial cell binding protein for high molecular weight kininogen and factor XII: identity with the receptor that binds to the globular "heads" of C1q (gC1q-R). AB - High molecular weight kininogen (HK) and factor XII are known to bind to human umbilical vein endothelial cells (HUVEC) in a zinc-dependent and saturable manner indicating that HUVEC express specific binding site(s) for those proteins. However, identification and immunochemical characterization of the putative receptor site(s) has not been previously accomplished. In this report, we have identified a cell surface glycoprotein that is a likely candidate for the HK binding site on HUVECs. When solubilized HUVEC membranes were subjected to an HK affinity column in the presence or absence of 50 microM ZnCl2 and the bound membrane proteins eluted, a single major protein peak was obtained only in the presence of zinc. SDS/PAGE analysis and silver staining of the protein peak revealed this protein to be 33 kDa and partial sequence analysis matched the NH2 terminus of gC1q-R, a membrane glycoprotein that binds to the globular "heads" of C1q. Two other minor proteins of approximately 70 kDa and 45 kDa were also obtained. Upon analysis by Western blotting, the 33-kDa band was found to react with several monoclonal antibodies (mAbs) recognizing different epitopes on gC1q R. Ligand and dot blot analyses revealed zinc-dependent binding of biotinylated HK as well as biotinylated factor XII to the isolated 33-kDa HUVEC molecule as well as recombinant gC1q-R. In addition, binding of 125I-HK to HUVEC cells was inhibited by selected monoclonal anti-gC1q-R antibodies. C1q, however, did not inhibit 125I-HK binding to HUVEC nor did those monoclonals known to inhibit C1q binding to gC1q-R. Taken together, the data suggest that HK (and factor XII) bind to HUVECs via a 33-kDa cell surface glycoprotein that appears to be identical to gC1q-R but interact with a site on gC1q-R distinct from that which binds C1q. PMID- 8710909 TI - Thymus transplantation, a critical factor for correction of autoimmune disease in aging MRL/+mice. AB - MRL/MP-+/+ (MRL/+) mice develop pancreatitis and sialoadenitis after they reach 7 months of age. Conventional bone marrow transplantation has been found to be ineffective in the treatment of these forms of apparent autoimmune disease. Old MRL/+ mice show a dramatic thymic involution with age. Hematolymphoid reconstitution is incomplete when fetal liver cells (as a source of hemopoietic stem cells) plus fetal bone (FB; which is used to recruit stromal cells) are transplanted from immunologically normal C57BL/6 donor mice to MRL/+ female recipients. Embryonic thymus from allogeneic C57BL/6 donors was therefore engrafted along with either bone marrow or fetal hematopoietic cells (FHCs) plus fragments of adult or fetal bone. More than seventy percent of old MRL/+ mice (> 7 months) that had been given a fetal thymus (FT) transplant plus either bone marrow or FHCs and also bone fragments survived more than 100 days after treatment. The mice that received FHCs, FB, plus FT from allogeneic donors developed normal T cell and B cell functions. Serum amylase levels decreased in these mice whereas they increased in the mice that received FHCs and FB but not FT. The pancreatitis and sialoadenitis already present at the time of transplantations were fully corrected according to histological analysis by transplants of allogeneic FHCs, FB and FT in the MRL/+ mice. These findings are taken as an experimental indication that perhaps stem cell transplants along with FT grafts might represent a useful strategy for treatment of autoimmune diseases in aged humans. PMID- 8710910 TI - Amelioration of lupus-like autoimmune disease in NZB/WF1 mice after treatment with a blocking monoclonal antibody specific for complement component C5. AB - New Zealand black x New Zealand white (NZB/W) F1 mice spontaneously develop an autoimmune syndrome with notable similarities to human systemic lupus erythematosus. Female NZB/WF1 mice produce high titers of antinuclear antibodies and invariably succumb to severe glomerulonephritis by 12 months of age. Although the development of the immune-complex nephritis is accompanied by abundant local and systemic complement activation, the role of proinflammatory complement components in disease progression has not been established. In this study we have examined the contribution of activated terminal complement proteins to the pathogenesis of the lupus-like autoimmune disease. Female NZB/W F1 mice were treated with a monoclonal antibody (mAb) specific for the C5 component of complement that blocks the cleavage of C5 and thus prevents the generation of the potent proinflammatory factors C5a and C5b-9. Continuous therapy with anti-C5 mAb for 6 months resulted in significant amelioration of the course of glomerulonephritis and in markedly increased survival. These findings demonstrate an important role for the terminal complement cascade in the progression of renal disease in NZB/W F1 mice, and suggest that mAb-mediated C5 inhibition may be a useful approach to the therapy of immune-complex glomerulonephritis in humans. PMID- 8710911 TI - A common major histocompatibility complex class II allele HLA-DQB1* 0301 is present in clinical variants of pemphigoid. AB - Bullous pemphigoid (BP) is an autoimmune subepidermal blistering disease seen primarily in elderly persons. It is characterized clinically by the development of tense bullae and by the presence of an antibasement membrane antibody. In BP, the antigens involved in the autoimmunity are epidermal basement membrane peptides BPAg1 and BPAg2. We have compared high resolution typing of major histocompatibility complex class II loci (HLA-DRB1, DQB1) in 21 patients with BP, 17 with ocular cicatricial pemphigoid (OCP), and 22 with oral pemphigoid (OP) to a panel of 218 haplotypes of normal individuals. We found that the three diseases (BP, OCP, and OP) have significant association with DQB1*0301 (P = 0.005, P < 0.0001, and P = 0.001, respectively). The frequencies of alleles DQB1*0302, 0303, and 06, which share a specific amino acid sequence from position 71 to 77 (Thr Arg-Ala-Glu-Leu-Val-Thr) were also increased (P = 0.01). We suggest that an identical major histocompatibility complex class II allele (DQB1*0301) is a common marker for enhanced susceptibility and that the same amino acid residues in positions 71-77 (DQB1*0301, -0302, -0305, -0602, -0603 alleles) are found in patients with BP, OCP and OP. Our findings propose that the autoimmune response in the three different clinical variants of pemphigoid, involves the recognition by T cells of a class II region of DQB1, bound to a peptide from the basement membrane of conjunctiva, oral mucosa, and skin. PMID- 8710912 TI - The proteolytic fragments generated by vertebrate proteasomes: structural relationships to major histocompatibility complex class I binding peptides. AB - Proteasomes are involved in the proteolytic generation of major histocompatibility complex (MHC) class I epitopes but their exact role has not been elucidated. We used highly purified murine 20S proteasomes for digestion of synthetic 22-mer and 41/44-mer ovalbumin partial sequences encompassing either an immunodominant or a marginally immunogenic epitope. At various times, digests were analyzed by pool sequencing and by semiquantitative electrospray ionization mass spectrometry. Most dual cleavage fragments derived from 22-mer peptides were 7-10 amino acids long, with octa- and nonamers predominating. Digestion of 41/44 mer peptides initially revealed major cleavage sites spaced by two size ranges, 8 or 9 amino acids and 14 or 15 amino acids, followed by further degradation of the latter as well as of larger single cleavage fragments. The final size distribution was slightly broader than that of fragments derived from 22-mer peptides. The majority of peptide bonds were cleaved, albeit with vastly different efficiencies. This resulted in multiple overlapping proteolytic fragments including a limited number of abundant peptides. The immunodominant epitope was generated abundantly whereas only small amounts of the marginally immunogenic epitope were detected. The frequency distributions of amino acids flanking proteasomal cleavage sites are correlated to that reported for corresponding positions of MHC class I binding peptides. The results suggest that proteasomal degradation products may include fragments with structural properties similar to MHC class I binding peptides. Proteasomes may thus be involved in the final stages of proteolytic epitope generation, often without the need for downstream proteolytic events. PMID- 8710913 TI - Induction of cytotoxic T lymphocytes by intramuscular immunization with plasmid DNA is facilitated by bone marrow-derived cells. AB - Striated muscle is the predominant site of gene expression after i.m. immunization of plasmid DNA, but it is not clear if myocytes or professional antigen-presenting cells (APCs) of hematopoietic origin present the encoded antigens to class I major histocompatibility complex (MHC)-restricted cytotoxic T lymphocytes (CTL). To address this issue, CTL responses were assessed in mice engrafted with immune systems that were partially MHC matched with antigen producing muscle cells. Spleen cells (sc) from immunocompetent F1 H-2bxd mice were infused into H-2b or H-2d mice carrying the severe combined immunodeficiency (scid) mutation, creating F1sc-->H-2b and F1sc-->H-2d chimeras, respectively. Immunization with DNA plasmids encoding the herpes simplex virus gB or the human immunodeficiency virus gp120 glycoproteins elicited antiviral CTL activity. F1sc- >H-2d chimeras responded to an H-2d-restricted gp120 epitope but not an H-2b restricted gB epitope, whereas F1sc-->H-2b chimeras responded to the H-2b but not the H-2d restricted epitope. This pattern of epitope recognition by the sc chimeras indicated that APCs of recipient (scid) origin were involved in initiation of CTL responses. Significantly, CTL responses against epitopes presented by the mismatched donor class I molecules were elicited if F1 bone marrow cells and sc were transferred into scid recipients before or several days to weeks after DNA immunization. Thus, bone marrow-derived APCs are sufficient for class I MHC presentation of viral antigens after i.m. immunization with plasmid DNA. Expression of plasmid DNA by these APCs is probably not a requirement for CTL priming. Instead, they appear to present proteins synthesized by other host cells. PMID- 8710916 TI - Insulin-secreting non-islet cells are resistant to autoimmune destruction. AB - Transgenic nonobese diabetic mice were created in which insulin expression was targeted to proopiomelanocortin-expressing pituitary cells. Proopiomelanocortin expressing intermediate lobe pituitary cells efficiently secrete fully processed, mature insulin via a regulated secretory pathway, similar to islet beta cells. However, in contrast to the insulin-producing islet beta cells, the insulin producing intermediate lobe pituitaries are not targeted or destroyed by cells of the immune system. Transplantation of the transgenic intermediate lobe tissues into diabetic nonobese diabetic mice resulted in the restoration of near normoglycemia and the reversal of diabetic symptoms. The absence of autoimmunity in intermediate lobe pituitary cells engineered to secrete bona fide insulin raises the potential of these cell types for beta-cell replacement therapy for the treatment of insulin-dependent diabetes mellitus. PMID- 8710915 TI - Loss of sequences 3' to the testis-determining gene, SRY, including the Y pseudoautosomal boundary associated with partial testicular determination. AB - The condition termed 46,XY complete gonadal dysgenesis is characterized by a completely female phenotype and streak gonads. In contrast, subjects with 46,XY partial gonadal dysgenesis and those with embryonic testicular regression sequence usually present ambiguous genitalia and a mix of Mullerian and Wolffian structures. In 46,XY partial gonadal dysgenesis gonadal histology shows evidence of incomplete testis determination. In 46,XY embryonic testicular regression sequence there is lack of gonadal tissue on both sides. Various lines of evidence suggest that embryonic testicular regression sequence is a variant form of 46,XY gonadal dysgenesis. The sex-determining region Y chromosome gene (SRY) encodes sequences for the testis-determining factor. To date germ-line mutations in SRY have been reported in approximately 20% of subjects with 46,XY complete gonadal dysgenesis. However, no germ-line mutations of SRY have been reported in subjects with the partial forms. We studied 20 subjects who presented either 46,XY partial gonadal dysgenesis or 46,XY embryonic testicular regression sequence. We examined the SRY gene and the minimum region of Y-specific DNA known to confer a male phenotype. The SRY-open reading frame (ORF) was normal in all subjects. However a de novo interstitial deletion 3' to the SRY-ORF was found in one subject. Although it is possible that the deletion was unrelated to the subject's phenotype, we propose that the deletion was responsible for the abnormal gonadal development by diminishing expression of SRY. We suggest that the deletion resulted either in the loss of sequences necessary for normal SRY expression or in a position effect that altered SRY expression. This case provides further evidence that deletions of the Y chromosome outside the SRY-ORF can result in either complete or incomplete sex reversal. PMID- 8710914 TI - Perturbations in maturation of secretory proteins and their association with endoplasmic reticulum chaperones in a cell culture model for epithelial ischemia. AB - The effects of ischemia on the maturation of secretory proteins are not well understood. Among several events that occur during ischemia-reperfusion are a rapid and extensive decrease in ATP levels and an alteration of cellular oxidative state. Since the normal folding and assembly of secretory proteins are mediated by endoplasmic reticulum (ER) molecular chaperones, the function of which depends on ATP and maintenance of an appropriate redox environment, ischemia might be expected to perturb folding of secretory proteins. In this study, whole animal and cultured cell models for the epithelial ischemic state were used to examine this possibility. After acute kidney ischemia, marked increases in the mRNA levels of the ER chaperones glucose-regulated protein (grp)78/immunoglobulin-binding protein (BiP), grp94, and ER protein (ERp)72 were noted. Likewise, when cellular ATP was depleted to less than 10% of control with antimycin A, mRNA levels of BiP, ERp72, and grp94 were increased in kidney and thyroid epithelial cell culture models. Since the signal for the up-regulation of these stress proteins is believed to be the accumulation of misfolded/misassembled secretory proteins in the ER, their induction after ischemia in vivo and antimycin treatment of cultured cells suggests that maturation of secretory proteins in the ER lumen might indeed be perturbed. To analyze the effects of antimycin A on the maturation of secretory proteins, we studied the fate of thyroglobulin (Tg), a large oligomeric secretory glycoprotein, the folding and assembly of which seems to require a variety of ER chaperones. Treatment of cultured thyroid epithelial cells with antimycin A greatly inhibited ( > 90%) the secretion of Tg. Sucrose density gradient analysis revealed that in antimycin A-treated cells Tg associates into large macromolecular complexes which, by immunofluorescence, appeared to localize to the ER. Furthermore, coimmunoprecipitation studies after antimycin A treatment demonstrated that Tg stably associates with BiP, grp94, and ERp72. Together, our results suggest that a key cellular lesion in ischemia is the misfolding of secretory proteins as they transit the ER, and this leads not only to increased expression of ER chaperones but also to their stable association with and the subsequent retention of at least some misfolded secretory proteins. PMID- 8710917 TI - Mapping rat megalin: the second cluster of ligand binding repeats contains a 46 amino acid pathogenic epitope involved in the formation of immune deposits in Heymann nephritis. AB - Megalin (gp330), an epithelial endocytic receptor, is a major target antigen of Heymann nephritis (HN), an autoimmune disease in rats. To elucidate the mechanisms of HN, we have mapped a pathogenic epitope in megalin that binds anti megalin antibodies. We focused our attention on four clusters of cysteine-rich, low density lipoprotein receptor (LDLR) ligand binding repeats in the extracellular domain of megalin because they represent putative ligand binding regions and therefore would be expected to be exposed in vivo and to be able to bind circulating antibodies. Rat megalin cDNA fragments I through IV encoding the first through fourth clusters of ligand-binding repeats, respectively, were expressed in a baculovirus system. All four expression products were detected by immunoblotting with two antisera capable of inducing passive HN (pHN). When antibodies eluted from glomeruli of rats with pHN were used for immunoblotting, only the expression product encoded by fragment II was detected. This indicates that the second cluster of LDLR ligand binding repeats is directly involved in binding anti-megalin antibodies and in the induction of pHN. To narrow the major epitope in this domain, fragment II was used to prepare proteins sequentially truncated from the C- and N-terminal ends by in vitro translation. Analysis of the truncated translation products by immunoprecipitation with anti-megalin IgG revealed that the fifth ligand-binding repeat (amino acids 1160-1205) contains the major epitope recognized. This suggests that a 46-amino acid sequence in the second cluster of LDLR ligand binding repeats contains a major pathogenic epitope that plays a key role in pHN. Identification of this epitope will facilitate studies on the pathogenesis of HN. PMID- 8710918 TI - A monoclonal IgG anticardiolipin antibody from a patient with the antiphospholipid syndrome is thrombogenic in mice. AB - Antiphospholipid antibodies, including anticardiolipin antibodies (ACA), are strongly associated with recurrent thrombosis in patients with the antiphospholipid syndrome (APS). To date, reports about the binding specificities of ACA and their role(s) in causing and/or sustaining thrombosis in APS are conflicting and controversial. The plasmas of patients with APS, usually containing a mixture of autoantibodies, vary in binding specificity for different phospholipids/cofactors and vary in in vitro lupus anticoagulant activity. Although in vivo assays that allow assessment of the pathogenic procoagulant activity of patient autoantibodies have recently been developed, the complex nature of the mixed species prevented determination of the particular species responsible for in vivo thrombosis. We have generated two human IgG monoclonal ACA from an APS patient with recurrent thrombosis. Both bound to cardiolipin in the presence of 10% bovine serum, but not in its absence, and both were reactive against phosphatidic acid, but were nonreactive against purified human beta-2 glycoprotein 1, DNA, heparan sulfate, or four other test antigens. Both monoclonal autoantibodies lacked lupus anticoagulant activity and did not inhibit prothrombinase activity. Remarkably, one of the monoclonal antibodies has thrombogenic properties when tested in an in vivo mouse model. This finding provides the first direct evidence that a particular antiphospholipid antibody specificity may contribute to in vivo thrombosis. PMID- 8710919 TI - Megalin-mediated endocytosis of transcobalamin-vitamin-B12 complexes suggests a role of the receptor in vitamin-B12 homeostasis. AB - Kidney cortex is a main target for circulating vitamin B12 (cobalamin) in complex with transcobalamin (TC). Ligand blotting of rabbit kidney cortex with rabbit 125I-TC-B12 and human TC-57Co-B12 revealed an exclusive binding to megalin, a 600 kDa endocytic receptor present in renal proximal tubule epithelium and other absorptive epithelia. The binding was Ca2+ dependent and inhibited by receptor associated protein (RAP). Surface plasmon resonance analysis demonstrated a high affinity interaction between purified rabbit megalin and rabbit TC-B12 but no measurable affinity of the vitamin complex for the homologous alpha 2 macroglobulin receptor (alpha 2MR)/low density lipoprotein receptor related protein (LRP). 125I-TC-B12 was efficiently endocytosed in a RAP-inhibitable manner in megalin-expressing rat yolk sac carcinoma cells and in vivo microperfused rat proximal tubules. The radioactivity in the tubules localized to the endocytic compartments and a similar apical distribution in the proximal tubules was demonstrated after intravenous injection of 125I-TC-B12. The TC-B12 binding sites in the proximal tubule epithelium colocalized with megalin as shown by ligand binding to cryosections of rat kidney cortex, and the binding was inhibited by anti-megalin polyclonal antibody, EDTA, and RAP. These data show a novel nutritional dimension of megalin as a receptor involved in the cellular uptake of vitamin B12. The expression of megalin in absorptive epithelia in the kidney and other tissues including yolk sac and placenta suggests a role of the receptor in vitamin B12 homeostasis and fetal vitamin B12 supply. PMID- 8710920 TI - Eradication of large colon tumor xenografts by targeted delivery of maytansinoids. AB - The maytansinoid drug DM1 is 100- to 1000-fold more cytotoxic than anticancer drugs that are currently in clinical use. The immunoconjugate C242-DM1 was prepared by conjugating DM1 to the monoclonal antibody C242, which recognizes a mucin-type glycoprotein expressed to various extents by human colorectal cancers. C242-DM1 was found to be highly cytotoxic toward cultured colon cancer cells in an antigen-specific manner and showed remarkable antitumor efficacy in vivo. C242 DM1 cured mice bearing subcutaneous COLO 205 human colon tumor xenografts (tumor size at time of treatment 65-130 mm3), at doses that showed very little toxicity and were well below the maximum tolerated dose. C242-DM1 could even effect complete regressions or cures in animals with large (260- to 500-mm3) COLO 205 tumor xenografts. Further, C242-DM1 induced complete regressions of subcutaneous LoVo and HT-29 colon tumor xenografts that express the target antigen in a heterogeneous manner. C242-DM1 represents a new generation of immunoconjugates that may yet fulfill the promise of effective cancer therapy through antibody targeting of cytotoxic agents. PMID- 8710921 TI - Characterization of the osmotic response element of the human aldose reductase gene promoter. AB - Aldose reductase (EC 1.1.1.21) catalyzes the NADPH-mediated conversion of glucose to sorbitol. The hyperglycemia of diabetes increases sorbitol production primarily through substrate availability and is thought to contribute to the pathogenesis of many diabetic complications. Increased sorbitol production can also occur at normoglycemic levels via rapid increases in aldose reductase transcription and expression, which have been shown to occur upon exposure of many cell types to hyperosmotic conditions. The induction of aldose reductase transcription and the accumulation of sorbitol, an organic osmolyte, have been shown to be part of the physiological osmoregulatory mechanism whereby renal tubular cells adjust to the intraluminal hyperosmolality during urinary concentration. Previously, to explore the mechanism regulating aldose reductase levels, we partially characterized the human aldose reductase gene promoter present in a 4.2-kb fragment upstream of the transcription initiation start site. A fragment (-192 to +31 bp) was shown to contain several elements that control the basal expression of the enzyme. In this study, we examined the entire 4.2-kb human AR gene promoter fragment by deletion mutagenesis and transfection studies for the presence of osmotic response enhancer elements. An 11-bp nucleotide sequence (TGGAAAATTAC) was located 3.7 kb upstream of the transcription initiation site that mediates hypertonicity-responsive enhancer activity. This osmotic response element (ORE) increased the expression of the chloramphenicol acetyltransferase reporter gene product 2-fold in transfected HepG2 cells exposed to hypertonic NaCl media as compared with isoosmotic media. A more distal homologous sequence is also described; however, this sequence has no osmotic enhancer activity in transfected cells. Specific ORE mutant constructs, gel shift, and DNA fragment competition studies confirm the nature of the element and identify specific nucleotides essential for enhancer activity. A plasmid construct containing three repeat OREs and a heterologous promoter increased expression 8-fold in isoosmotic media and an additional 4-fold when the transfected cells are subjected to hyperosmotic stress (total approximately 30 fold). These findings will permit future studies to identify the transcription factors involved in the normal regulatory response mechanism to hypertonicity and to identify whether and how this response is altered in a variety of pathologic states, including diabetes. PMID- 8710922 TI - Overexpression of insulin-like growth factor-1 in the heart is coupled with myocyte proliferation in transgenic mice. AB - Transgenic mice were generated in which the cDNA for the human insulin-like growth factor 1B (IGF-1B) was placed under the control of a rat alpha-myosin heavy chain promoter. In mice heterozygous for the transgene, IGF-1B mRNA was not detectable in the fetal heart at the end of gestation, was present in modest levels at 1 day after birth, and increased progressively with postnatal maturation, reaching a peak at 75 days. Myocytes isolated from transgenic mice secreted 1.15 +/- 0.25 ng of IGF-1 per 10(6) cells per 24 hr versus 0.27 +/- 0.10 ng in myocytes from homozygous wild-type littermates. The plasma level of IGF-1 increased 84% in transgenic mice. Heart weight was comparable in wild-type littermates and transgenic mice up to 45 days of age, but a 42%, 45%, 62%, and 51% increase was found at 75, 135, 210, and 300 days, respectively, after birth. At 45, 75, and 210 days, the number of myocytes in the heart was 21%, 31%, and 55% higher, respectively, in transgenic animals. In contrast, myocyte cell volume was comparable in transgenic and control mice at all ages. In conclusion, overexpression of IGF-1 in myocytes leads to cardiomegaly mediated by an increased number of cells in the heart. PMID- 8710923 TI - Low molecular weight EPS II of Rhizobium meliloti allows nodule invasion in Medicago sativa. AB - Effective invasion of alfalfa by Rhizobium meliloti Rm1021 normally requires the presence of succinoglycan, an exopolysaccharide (EPS) produced by the bacterium. However, Rm1021 has the ability to produce a second EPS (EPS II) that can suppress the symbiotic defects of succinoglycan-deficient strains. EPS II is a polymer of modified glucose-(beta-1,3)-galactose subunits and is produced by Rm1021 derivatives carrying either an expR101 or mucR mutation. If the ability to synthesize succinoglycan is blocked genetically, expR101 derivatives of Rm1021 are nodulation-proficient, whereas mucR derivatives of Rm1021 are not. The difference in nodulation proficiency between these two classes of EPS II producing strains is due to the specific production of a low molecular weight form of EPS II by expR101 strains. A low molecular weight EPS II fraction consisting of 15-20 EPS II disaccharide subunits efficiently allows nodule invasion by noninfective strains when present in amounts as low as 7 pmol per plant, suggesting that low molecular weight EPS II may act as a symbiotic signal during infection. PMID- 8710924 TI - Haemonchus contortus GA1 antigens: related, phospholipase C-sensitive, apical gut membrane proteins encoded as a polyprotein and released from the nematode during infection. AB - It was previously shown that the Haemonchus contortus apical gut surface proteins p46, p52, and p100 induced protective immunity to challenge infections in goats. Here, it is shown that the three proteins are all encoded by a single gene (GA1) and initially expressed in adult parasites as a polyprotein (p100GA1). p46GA1 and p52GA1 are related proteins with 47% sequence identity, including a cysteine containing region, which appears to confer secondary structure to these proteins, and a region with sequence similarity to bacterial Tolb proteins. GA1 protein expression is regulated during the life cycle at the level of transcript abundance. Only p52GA1 has characteristics of a glycosylinositolphospholipid membrane-anchored protein. However, both p46GA1 and p52GA1 were released from the gut membrane by phosphatidylinositol specific-phospholipase C, suggesting that p46GA1 membrane association depends on interactions with a glycosylinositolphospholipid gut membrane protein. Finally, GA1 proteins occur in abomasal mucus of infected lambs, demonstrating possible presentation to the host immune system during H. contortus infection. The results identify multiple characteristics of the GA1 proteins that should be considered for design of recombinant antigens for vaccine trials and that implicate a series of cellular processes leading to modification and expression of GA1 proteins at the nematode apical gut surface. PMID- 8710925 TI - Probing the pore region of recombinant N-methyl-D-aspartate channels using external and internal magnesium block. AB - Mg2+ ions block N-methyl-D-aspartate (NMDA) channels by entering the pore from either the extracellular or the cytoplasmic side of the membrane in a voltage dependent manner. We have used these two different block phenomena to probe the structure of the subunits forming NMDA channels. We have made several amino acid substitutions downstream of the Q/R/N site in the TMII region of both NR1 and NR2A subunits. Mutant NR1 subunits were coexpressed with wild-type NR2A subunits and vice versa in Xenopus oocytes. We found that individually mutating the first two amino acid residues downstream to the Q/R/N site affects mostly the block by external Mg2+. Mutations of residues five to seven positions downstream of the Q/R/N site do not influence the external Mg2+ block, but clearly influence the block by internal Mg2+. These data add support to the hypothesis that there are two separate binding sites for external and internal Mg2+ block. They also indicate that the C-terminal end of TMII contributes to the inner vestibule of the pore of NMDA channels and thus provide additional evidence that TMII forms a loop that reemerges toward the cytoplasmic side of the membrane. PMID- 8710926 TI - Locust adipokinetic hormones: carrier-independent transport and differential inactivation at physiological concentrations during rest and flight. AB - Since concomitant release of structurally related peptide hormones with apparently similar functions seems to be a general concept in endocrinology, we have studied the dynamics of the lifetime of the three known adipokinetic hormones (AKHs) of the migratory locust, which control flight-directed mobilization of carbohydrate and lipid from fat body stores. Although the structure of the first member of the AKHs has been known for 20 years, until now, reliable data on their inactivation and removal from the hemolymph are lacking, because measurement requires AKHs with high specific radioactivity. Employing tritiated AKHs with high specific radioactivity, obtained by catalytic reduction with tritium gas of the dehydroLeu2 analogues of the AKHs synthesized by the solid-phase procedure, studies with physiological doses of as low as 1.0 pmol per locust could be conducted. The AKHs appear to be transported in the hemolymph in their free forms and not associated with a carrier protein, despite their strong hydrophobicity. Application of AKHs in their free form in in vivo and in vitro studies therefore now has been justified. We have studied the degradation of the three AKHs during rest and flight. The first cleavage step by an endopeptidase is crucial, since the resulting degradation products lack any adipokinetic activity. Half-lives for AKH-I, -II and -III were 51, 40, and 5 min, respectively, for rest conditions and 35, 37, and 3 min, respectively, during flight. The rapid and differential degradation of structurally related hormones leads to changes in the ratio in which they are released and therefore will have important consequences for concerted hormone action at the level of the target organ or organs, suggesting that each of the known AKHs may play its own biological role in the overall syndrome of insect flight. PMID- 8710927 TI - The hippocampal formation participates in novel picture encoding: evidence from functional magnetic resonance imaging. AB - Considerable evidence exists to support the hypothesis that the hippocampus and related medial temporal lobe structures are crucial for the encoding and storage of information in long-term memory. Few human imaging studies, however, have successfully shown signal intensity changes in these areas during encoding or retrieval. Using functional magnetic resonance imaging (fMRI), we studied normal human subjects while they performed a novel picture encoding task. High-speed echo-planar imaging techniques evaluated fMRI signal changes throughout the brain. During the encoding of novel pictures, statistically significant increases in fMRI signal were observed bilaterally in the posterior hippocampal formation and parahippocampal gyrus and in the lingual and fusiform gyri. To our knowledge, this experiment is the first fMRI study to show robust signal changes in the human hippocampal region. It also provides evidence that the encoding of novel, complex pictures depends upon an interaction between ventral cortical regions, specialized for object vision, and the hippocampal formation and parahippocampal gyrus, specialized for long-term memory. PMID- 8710928 TI - Structure, tissue distribution, and chromosomal localization of the prepronociceptin gene. AB - Nociceptin (orphanin FQ), the newly discovered natural agonist of opioid receptor like (ORL1) receptor, is a neuropeptide that is endowed with pronociceptive activity in vivo. Nociceptin is derived from a larger precursor, prepronociceptin (PPNOC), whose human, mouse, and rat genes we have now isolated. The PPNOC gene is highly conserved in the three species and displays organizational features that are strikingly similar to those of the genes of preproenkephalin, preprodynorphin, and preproopiomelanocortin, the precursors to endogenous opioid peptides, suggesting the four genes belong to the same family-i.e., have a common evolutionary origin. The PPNOC gene encodes a single copy of nociceptin as well as of other peptides whose sequence is strictly conserved across murine and human species; hence it is likely to be neurophysiologically significant. Northern blot analysis shows that the PPNOC gene is predominantly transcribed in the central nervous system (brain and spinal cord) and, albeit weakly, in the ovary, the sole peripheral organ expressing the gene. By using a radiation hybrid cell line panel, the PPNOC gene was mapped to the short arm of human chromosome 8 (8p21), between sequence-tagged site markers WI-5833 and WI-1172, in close proximity of the locus encoding the neurofilament light chain NEFL. Analysis of yeast artificial chromosome clones belonging to the WC8.4 contig covering the 8p21 region did not allow to detect the presence of the gene on these yeast artificial chromosomes, suggesting a gap in the coverage within this contig. PMID- 8710929 TI - Norepinephrine transporters have channel modes of conduction. AB - Neurotransmitter transporters couple to existing ion gradients to achieve reuptake of transmitter into presynaptic terminals. For coupled cotransport, substrates and ions cross the membrane in fixed stoichiometry. This is in contrast to ion channels, which carry an arbitrary number of ions depending on the channel open time. Members of the gamma-aminobutyric acid transporter gene family presumably function with fixed stoichiometry in which a set number of ions cotransport with one transmitter molecule. Here we report channel-like events from a presumably fixed stoichiometry [norepinephrine (NE)+, Na+, and Cl-], human NE (hNET) in the gamma-aminobutyric acid transporter gene family. These events are stimulated by NE and by guanethidine, an hNET substrate, and they are blocked by cocaine and the antidepressant desipramine. Voltage-clamp data combined with NE uptake data from these same cells indicate that hNETs have two functional modes of conduction: a classical transporter mode (T-mode) and a novel channel mode (C-mode). Both T-mode and C-mode are gated by the same substrates and antagonized by the same blockers. T-mode is putatively electrogenic because the transmitter and cotransported ions sum to one net charge. However, C-mode carries virtually all of the transmitter-induced current, even though it occurs with low probability. This is because each C-mode opening transports hundreds of charges per event. The existence of a channel mode of conduction in a previously established fixed-stoichiometry transporter suggests the appearance of an aqueous pore through the transporter protein during the transport cycle and may have significance for transporter regulation. PMID- 8710930 TI - Primary structure and tissue distribution of the orphanin FQ precursor. AB - The heptadecapeptide orphanin FQ (OFQ) is a recently discovered neuropeptide that exhibits structural features reminiscent of the opioid peptides and that is an endogenous ligand to a G protein-coupled receptor sequentially related to the opioid receptors. We have cloned both the human and rat cDNAs encoding the OFQ precursor proteins, to investigate whether the sequence relationships existing between the opioid and OFQ systems are also found at the polypeptide precursor level, in particular whether the OFQ precursor would encode several bioactive peptides as do the opioid precursors, and to study the regional distribution of OFQ sites of synthesis. The entire precursor protein displays structural homology to the opioid peptide precursors, especially preprodynorphin and preproenkephalin. The predicted amino acid sequence of the OFQ precursor contains a putative signal peptide and one copy of the OFQ sequence flanked by pairs of basic amino acid residues. Carboxyl-terminal to the OFQ sequence, the human and rat precursors contain a stretch of 28 amino acids that is 100% conserved and thus may encode novel bioactive peptides. Two peptides derived from this stretch were synthesized but were found to be unable to activate the OFQ receptor, suggesting that if they are produced in vivo, these peptides would likely recognize receptors different from the OFQ receptor. To begin analyzing the sites of OFQ mRNA synthesis, Northern analysis of human and rat tissues were carried out and showed that the OFQ precursor mRNA is mainly expressed in the brain. In situ hybridization of rat brain slices demonstrated a regional distribution pattern of the OFQ precursor mRNA, which is distinct from that of the opioid peptide precursors. These data confirm that the OFQ system differs from the opioid system at the molecular level, although the OFQ and opioid precursors may have arisen from a common ancestral gene. PMID- 8710931 TI - The temporal lobe is a target of output from the basal ganglia. AB - The basal ganglia are known to receive inputs from widespread regions of the cerebral cortex, such as the frontal, parietal, and temporal lobes. Of these cortical areas, only the frontal lobe is thought to be the target of basal ganglia output. One of the cortical regions that is a source of input to the basal ganglia is area TE, in inferotemporal cortex. This cortical area is thought to be critically involved in the recognition and discrimination of visual objects. Using retrograde transneuronal transport of herpes simplex virus type 1, we have found that one of the output nuclei of the basal ganglia, the substantia nigra pars reticulata, projects via the thalamus to TE. Thus, TE is not only a source of input to the basal ganglia, but also is a target of basal ganglia output. This result implies that the output of the basal ganglia influences higher order aspects of visual processing. In addition, we propose that dysfunction of the basal ganglia loop with TE leads to alterations in visual perception, including visual hallucinations. PMID- 8710932 TI - Surface protein phosphorylation by ecto-protein kinase is required for the maintenance of hippocampal long-term potentiation. AB - During the induction of long-term potentiation (LTP) in hippocampal slices adenosine triphosphate (ATP) is secreted into the synaptic cleft, and a 48 kDa/50 kDa protein duplex becomes phosphorylated by extracellular ATP. All the criteria required as evidence that these two proteins serve as principal substrates of ecto-protein kinase activity on the surface of hippocampal pyramidal neurons have been fulfilled. This phosphorylation activity was detected on the surface of pyramidal neurons assayed after synaptogenesis, but not in immature neurons nor in glial cells. Addition to the extracellular medium of a monoclonal antibody termed mAb 1.9, directed to the catalytic domain of protein kinase C (PKC), inhibited selectively this surface protein phosphorylation activity and blocked the stabilization of LTP induced by high frequency stimulation (HFS) in hippocampal slices. This antibody did not interfere with routine synaptic transmission nor prevent the initial enhancement of synaptic responses observed during the 1-5 min period immediately after the application of HFS (the induction phase of LTP). However, the initial increase in the slope of excitatory postsynaptic potentials, as well as the elevated amplitude of the population spike induced by HFS, both declined gradually and returned to prestimulus values within 30-40 min after HFS was applied in the presence of mAb 1.9. A control antibody that binds to PKC but does not inhibit its activity had no effect on LTP. The selective inhibitory effects observed with mAb 1.9 provide the first direct evidence of a causal role for ecto-PK in the maintenance of stable LTP, an event implicated in the process of learning and the formation of memory in the brain. PMID- 8710933 TI - Role for cells in the presupplementary motor area in updating motor plans. AB - Two motor areas are known to exist in the medial frontal lobe of the cerebral cortex of primates, the supplementary motor area (SMA) and the presupplementary motor area (pre-SMA). We report here on an aspect of cellular activity that characterizes the pre-SMA. Monkeys were trained to perform three different movements sequentially in a temporal order. The correct order was planned on the basis of visual information before its execution. A group of pre-SMA cells (n = 64, 25%) were active during a process when monkeys were required to discard a current motor plan and develop a plan appropriate for the next orderly movements. Such activity was not common in the SMA and not found in the primary motor cortex. Our data suggest a role of pre-SMA cells in updating motor plans for subsequent temporally ordered movements. PMID- 8710934 TI - Mice lacking the gene encoding tissue-type plasminogen activator show a selective interference with late-phase long-term potentiation in both Schaffer collateral and mossy fiber pathways. AB - The gene encoding tissue-type plasminogen activator (t-PA) is an immediate response gene, downstream from CREB-1 and other constitutively expressed transcription factors, which is induced in the hippocampus during the late phase of long-term potentiation (L-LTP). Mice in which the t-PA gene has been ablated (t-PA-/-) showed no gross anatomical, electrophysiological, sensory, or motor abnormalities but manifest a selective reduction in L-LTP in hippocampal slices in both the Schaffer collateral-CA1 and mossy fiber-CA3 pathways. t-PA-/- mice also exhibit reduced potentiation by cAMP analogs and D1/D5 agonists. By contrast, hippocampal-dependent learning and memory were not affected in these mice, whereas performance was impaired on two-way active avoidance, a striatum dependent task. These results provide genetic evidence that t-PA is a downstream effector gene important for L-LTP and show that modest impairment of L-LTP in CA1 and CA3 does not result in hippocampus-dependent behavioral phenotypes. PMID- 8710935 TI - Activation of muscarinic acetylcholine receptors via their allosteric binding sites. AB - Ligands that bind to the allosteric-binding sites on muscarinic acetylcholine receptors alter the conformation of the classical-binding sites of these receptors and either diminish or increase their affinity for muscarinic agonists and classical antagonists. It is not known whether the resulting conformational change also affects the interaction between the receptors and the G proteins. We have now found that the muscarinic receptor allosteric modulators alcuronium, gallamine, and strychnine (acting in the absence of an agonist) alter the synthesis of cAMP in Chinese hamster ovary (CHO) cells expressing the M2 or the M4 subtype of muscarinic receptors in the same direction as the agonist carbachol. In addition, most of their effects on the production of inositol phosphates in CHO cells expressing the M1 or the M3 muscarinic receptor subtypes are also similar to (although much weaker than) those of carbachol. The agonist like effects of the allosteric modulators are not observed in CHO cells that have not been transfected with the gene for any of the subtypes of muscarinic receptors. The effects of alcuronium on the formation of cAMP and inositol phosphates are not prevented by the classical muscarinic antagonist quinuclidinyl benzilate. These observations demonstrate for the first time that the G protein mediated functional responses of muscarinic receptors can be evoked not only from their classical, but also from their allosteric, binding sites. This represents a new mechanism of receptor activation. PMID- 8710936 TI - Long-term potentiation at single fiber inputs to hippocampal CA1 pyramidal cells. AB - Despite extensive investigation, it remains unclear whether presynaptic and/or postsynaptic modifications are primarily responsible for the expression of long term potentiation (LTP) in the CA1 region of the hippocampus. Here we address this issue by using techniques that maximize the likelihood of stimulating a single axon and thereby presumably a single synapse before and after the induction of LTP. Several basic properties of synaptic transmission were examined including the probability of neurotransmitter release (Pr), the quantal size (q), and the so-called potency, which is defined as the average size of the synaptic response when release of transmitter does occur. LTP was routinely associated with an increase in potency, whereas increases in Pr alone were not observed. LTP was also reliably induced when baseline Pr was high, indicating that synapses with high Pr can express LTP. These results suggest that the mechanism for the expression of LTP involves an increase in q and is difficult to explain by an increase in Pr alone. PMID- 8710937 TI - Glucocorticoids have state-dependent stimulant effects on the mesencephalic dopaminergic transmission. AB - An increase in the activity of mesencephalic dopaminergic neurons has been implicated in the appearance of pathological behaviors such as psychosis and drug abuse. Several observations suggest that glucocorticoids might contribute to such an increase in dopaminergic activity. The present experiments therefore analyzed the effects of corticosterone, the major glucocorticoid in the rat, both on dopamine release in the nucleus accumbens of freely moving animals by means of microdialysis, and on locomotor activity, a behavior dependent on accumbens dopamine. Given that glucocorticoids have certain state-dependent neuronal effects, their action on dopamine was studied in situations differing in dopaminergic tonus, including during the light and dark phases of the circadian cycle, during eating, and in groups of animals differing in their locomotor reactivity to novelty. Dopaminergic activity is increased in the dark period, further increased during food-intake, and is higher in rats defined as high responders to novelty than in low responders. Corticosterone, peripherally administered in a dose that approximates stress-induced plasma concentrations, increased extracellular concentrations of dopamine, and this increase was augmented in the dark phase, during eating, and in high responder rats. Corticosterone had little or no effects in the light phase and in low responder rats. Corticosterone also stimulated locomotor activity, an effect that paralleled the release of dopamine and was abolished by neurochemical (6 hydroxydopamine) depletion of accumbens dopamine. In conclusion, glucocorticoids have state-dependent stimulant effects on mesencephalic dopaminergic transmission, and an interaction between these two factors might be involved in the appearance of behavioral disturbances. PMID- 8710938 TI - Opposite actions of nitric oxide on cholinergic synapses: which pathways? AB - Nitric oxide (NO) produced opposite effects on acetylcholine (ACh) release in identified neuroneuronal Aplysia synapses depending on the excitatory or the inhibitory nature of the synapse. Extracellular application of the NO donor, SIN 1, depressed the inhibitory postsynaptic currents (IPSCs) and enhanced the excitatory postsynaptic currents (EPSCs) evoked by presynaptic action potentials (1/60 Hz). Application of a membrane-permeant cGMP analog mimicked the effect of SIN-1 suggesting the participation of guanylate cyclase in the NO pathway. The guanylate cyclase inhibitor, methylene blue, blocked the NO-induced enhancement of EPSCs but only reduced the inhibition of IPSCs indicating that an additional mechanism participates to the depression of synaptic transmission by NO. Using nicotinamide, an inhibitor of ADP-ribosylation, we found that the NO-induced depression of ACh release on the inhibitory synapse also involves ADP ribosylation mechanism(s). Furthermore, application of SIN-1 paired with cGMP dependent protein kinase (cGMP-PK) inhibitors showed that cGMP-PK could play a role in the potentiating but not in the depressing effect of NO on ACh release. Increasing the frequency of stimulation of the presynaptic neuron from 1/60 Hz to 0.25 or 1 Hz potentiated the EPSCs and reduced the IPSCs. In these conditions, the potentiating effect of NO on the excitatory synapse was reduced, whereas its depressing effect on the inhibitory synapse was unaffected. Moreover the frequency-dependent enhancement of ACh release in the excitatory synapse was greatly reduced by the inhibition of NO synthase. Our results indicate that NO may be involved in different ways of modulation of synaptic transmission depending on the type of the synapse including synaptic plasticity. PMID- 8710939 TI - Resonant tectorial membrane motion in the inner ear: its crucial role in frequency tuning. AB - The tectorial membrane has long been postulated as playing a role in the exquisite sensitivity of the cochlea. In particular, it has been proposed that the tectorial membrane provides a second resonant system, in addition to that of the basilar membrane, which contributes to the amplification of the motion of the cochlear partition. Until now, technical difficulties had prevented vibration measurements of the tectorial membrane and, therefore, precluded direct evidence of a mechanical resonance. In the study reported here, the vibration of the tectorial membrane was measured in two orthogonal directions by using a novel method of combining laser interferometry with a photodiode technique. It is shown experimentally that the motion of the tectorial membrane is resonant at a frequency of 0.5 octave (oct) below the resonant frequency of the basilar membrane and polarized parallel to the reticular lamina. It is concluded that the resonant motion of the tectorial membrane is due to a parallel resonance between the mass of the tectorial membrane and the compliance of the stereocilia of the outer hair cells. Moreover, in combination with the contractile force of outer hair cells, it is proposed that inertial motion of the tectorial membrane provides the necessary conditions to allow positive feedback of mechanical energy into the cochlear partition, thereby amplifying and tuning the cochlear response. PMID- 8710940 TI - Overview of the most prevalent hypothalamus-specific mRNAs, as identified by directional tag PCR subtraction. AB - We applied the directional tag PCR subtractive hybridization method to construct a rat hypothalamic cDNA library from which cerebellar and hippocampal sequences had been depleted, enriching 20-30-fold for sequences expressed selectively in the hypothalamus. We studied a sample of 94 clones selected for enrichment in the subtracted library. These clones corresponded to 43 distinct mRNA species, about half of which were novel. Thirty-eight of these 43 mRNAs (corresponding to 85 of the clones in the sample) exhibited enrichment in the hypothalamus; 23 were highly enriched. In situ hybridization studies revealed that one novel species was restricted to cells in a small bilaterally symmetric area of the paraventricular hypothalamus. Other novel mRNAs showed substantial enrichment in basal diencephalic structures, particularly the hypothalamus, without restriction to single hypothalamic nuclei. The data suggest that the hypothalamus utilizes at least two distinct strategies for employing its selectively expressed proteins. Secretory neuropeptides utilized for intercellular communication are produced by functionally discrete nuclei, while several other proteins are shared by structures that are unrelated in their physiological roles but may share biochemical systems. PMID- 8710941 TI - 16 alpha-substituted analogs of the antiprogestin RU486 induce a unique conformation in the human progesterone receptor resulting in mixed agonist activity. AB - Previously, we have shown that agonists and antagonists interact with distinct, though overlapping regions within the human progesterone receptor (hPR) resulting in the formation of structurally different complexes. Thus, a link was established between the structure of a ligand-receptor complex and biological activity. In this study, we have utilized a series of in vitro assays with which to study hPR pharmacology and have identified a third class of hPR ligands that induce a receptor conformation which is distinct from that induced by agonists or antagonists. Importantly, when assayed on PR-responsive target genes these compounds were shown to exhibit partial agonist activity; an activity that was influenced by cell context. Thus, as has been shown previously for estrogen receptor, the overall structure of the ligand-receptor complex is influenced by the nature of the ligand. It appears, therefore, that the observed differences in the activity of some PR and estrogen receptor ligands reflect the ability of the cellular transcription machinery to discriminate between the structurally different complexes that result following ligand interaction. These data support the increasingly favored hypothesis that different ligands can interact with different regions within the hormone binding domains of steroid hormone receptors resulting in different biologies. PMID- 8710942 TI - Circulating plasma xanthine oxidase contributes to vascular dysfunction in hypercholesterolemic rabbits. AB - Reactive oxygen species play a central role in vascular inflammation and atherogenesis, with enhanced superoxide (O2.-) production contributing significantly to impairment of nitric oxide (.NO)-dependent relaxation of vessels from cholesterol-fed rabbits. We investigated potential sources of O2.- production, which contribute to this loss of endothelium-dependent vascular responses. The vasorelaxation elicited by acetylcholine (ACh) in phenylephrine contracted, aortic ring segments was impaired by cholesterol feeding. Pretreatment of aortic vessels with either heparin, which competes with xanthine oxidase (XO) for binding to sulfated glycosaminoglycans, or the XO inhibitor allopurinol resulted in a partial restoration (36-40% at 1 muM ACh) of ACh dependent relaxation. Furthermore, O2.(-)-dependent lucigenin chemiluminescence, measured in intact ring segments from hypercholesterolemic rabbits, was decreased by addition of heparin, allopurinol or a chimeric, heparin-binding superoxide dismutase. XO activity was elevated more than two-fold in plasma of hypercholesterolemic rabbits. Incubation of vascular rings from rabbits on a normal diet with purified XO (10 milliunits/ml) also impaired .NO-dependent relaxation but only in the presence of purine substrate. As with vessels from hypercholesterolemic rabbits, this effect was prevented by heparin and allopurinol treatment. We hypothesize that increases in plasma cholesterol induce the release of XO into the circulation, where it binds to endothelial cell glycosaminoglycans. Only in hypercholesterolemic vessels is sufficient substrate available to sustain the production of O2.- and impair NO-dependent vasorelaxation. Chronically, the continued production of peroxynitrite, (ONOO-) which the simultaneous generation of NO and O2.- implies, may irreversibly impair vessel function. PMID- 8710943 TI - Overexpression of the transcription factor UBF1 is sufficient to increase ribosomal DNA transcription in neonatal cardiomyocytes: implications for cardiac hypertrophy. AB - The accelerated protein accumulation characteristic of cardiomyocyte hypertrophy results from increased cellular protein synthetic capacity (elevated ribosome content). The rate limiting step in ribosome accumulation is transcription of the rRNA genes. During neonatal cardiomyocyte hypertrophy induced by norepinephrine or spontaneous contraction, changes in the expression of a ribosomal DNA transcription factor, UBF, correlated with increased rates of ribosome biogenesis. We hypothesized that elevated expression of UBF was part of the mechanism by which these hypertrophic stimuli effected increases in the rate of transcription from the rDNA promoter. In this study, we have examined directly the effect of overexpressing UBF on rDNA transcription in neonatal cardiomyocytes in culture. In control experiments, a novel reporter construct for rDNA transcription (pSMECAT) showed similar increases in activity in response to hypertrophic stimuli (10(-4) M phenylephrine, 10(-7) M endothelin, and spontaneous contraction) as did the endogenous rRNA genes. When contraction arrested cardiomyocytes were cotransfected with pSMECAT and increasing amounts of a UBF1 expression vector; a dose-dependent (3-5 fold) increase in rDNA transcription was observed. Western blot analysis confirmed that the overexpressed, FLAG-tagged UBF accumulated in the cardiomyocyte nuclei. The observation that overexpression of UBF1 is sufficient to increase rDNA transcription in neonatal cardiomyocytes provides evidence in support of the hypothesis that the regulation of UBF is a key component of the increased ribosome biogenesis and protein accumulation associated with cardiomyocyte hypertrophy. PMID- 8710944 TI - Imaging ROMK1 inwardly rectifying ATP-sensitive K+ channel protein using atomic force microscopy. AB - The inwardly rectifying K+ channel ROMK1 has been implicated as being significant in K+ secretion in the distal nephron. ROMK1 has been shown by immunocytochemistry to be expressed in relevant nephron segments. The development of the atomic force microscope has made possible the production of high resolution images of small particles, including a variety of biological macromolecules. Recently, a fusion protein of glutathione S-transferase (GST) and ROMK1 (ROMK1-GST) has been used to produce a polyclonal antibody for immunolocalization of ROMK1. We have used atomic force microscopy to examine ROMK1-GST and the native ROMK1 polypeptide cleaved from GST. Imaging was conducted with the proteins in physiological solutions attached to mica. ROMK1 GST appears in images as a particle composed of two units of similar size. Analyses of images indicate that the two units have volumes of approximately 118 nm3, which is close to the theoretical volume of a globular protein of approximately 65 kDa (the molecular mass of ROMK1-GST). Native GST exists as a dimer, and the images obtained here are consistent with the ROMK1-GST fusion protein's existence as a heterodimer. In experiments on ROMK1 in aqueous solution, single molecules appear to aggregate, but contact to the mica was maintained. Addition of ATP to the solution produced a change in height of the aggregates. This change (which was reversible) suggests that ATP induces a structural change in the ROMK1 protein. The data show that atomic force microscopy is a useful tool for examination of purified protein molecules under near-physiological conditions, and furthermore, that structural alterations in the proteins may be continuously investigated. PMID- 8710945 TI - The physical and genomic organization of microsatellites in sugar beet. AB - Microsatellites, tandem arrays of short (2-5 bp) nucleotide motifs, are present in high numbers in most eukaryotic genomes. We have characterized the physical distribution of microsatellites on chromosomes of sugar beet (Beta vulgaris L.). Each microsatellite sequence shows a characteristic genomic distribution and motif-dependent dispersion, with site-specific amplification on one to seven pairs of centromeres or intercalary chromosomal regions and weaker, dispersed hybridization along chromosomes. Exclusion of some microsatellites from 18S-5.8S 25S rRNA gene sites, centromeres, and intercalary sites was observed. In-gel and in situ hybridization patterns are correlated, with highly repeated restriction fragments indicating major centromeric sites of microsatellite arrays. The results have implications for genome evolution and the suitability of particular microsatellite markers for genetic mapping and genome analysis. PMID- 8710946 TI - Creation of a novel protein-coding region at the RNA level in black pine chloroplasts: the pattern of RNA editing in the gymnosperm chloroplast is different from that in angiosperms. AB - The phenomenon of RNA editing has been found to occur in chloroplasts of several angiosperm plants. Comparative analysis of the entire nucleotide sequence of a gymnosperm [Pinus thunbergii (black pine)] chloroplast genome allowed us to predict several potential editing sites in its transcripts. Forty-nine such sites from 14 genes/ORFs were analyzed by sequencing both cDNAs from the transcripts and the corresponding chloroplast DNA regions, and 26 RNA editing sites were identified in the transcripts from 12 genes/ORFs, indicating that chloroplast RNA editing is not restricted to angiosperms but occurs in the gymnosperm, too. All the RNA editing events are C-to-U conversions; however, many new codon substitutions and creation of stop codons that have not so far been reported in angiosperm chloroplasts were observed. The most striking is that two editing events result in the creation of an initiation and a stop codon within a single transcript, leading to the formation of a new reading frame of 33 codons. The predicted product is highly homologous to that deduced from the ycf7 gene (ORF31), which is conserved in the chloroplast genomes of many other plant species. PMID- 8710947 TI - Expression of a delta 9 14:0-acyl carrier protein fatty acid desaturase gene is necessary for the production of omega 5 anacardic acids found in pest-resistant geranium (Pelargonium xhortorum). AB - Anacardic acids, a class of secondary compounds derived from fatty acids, are found in a variety of dicotyledonous families. Pest resistance (e.g., spider mites and aphids) in Pelargonium xhortorum (geranium) is associated with high levels (approximately 81%) of unsaturated 22:1 omega 5 and 24:1 omega 5 anacardic acids in the glandular trichome exudate. A single dominant locus controls the production of these omega 5 anacardic acids, which arise from novel 16:1 delta 11 and 18:1 delta 13 fatty acids. We describe the isolation and characterization of a cDNA encoding a unique delta 9 14:0-acyl carrier protein fatty acid desaturase. Several lines of evidence indicated that expression of this desaturase leads to the production of the omega 5 anacardic acids involved in pest resistance. First, its expression was found in pest-resistant, but not suspectible, plants and its expression followed the production of the omega 5 anacardic acids in segregating populations. Second, its expression and the occurrence of the novel 16:1 delta 11 and 18:1 delta 13 fatty acids and the omega 5 anacardic acids were specific to tall glandular trichomes. Third, assays of the recombinant protein demonstrated that this desaturase produced the 14:1 delta 9 fatty acid precursor to the novel 16:1 delta 11 and 18:1 delta 13 fatty acids. Based on our genetic and biochemical studies, we conclude that expression of this delta 9 14:0-ACP desaturase gene is required for the production of omega 5 anacardic acids that have been shown to be necessary for pest resistance in geranium. PMID- 8710948 TI - The N gene of tobacco confers resistance to tobacco mosaic virus in transgenic tomato. AB - It has been proposed that cloned plant disease resistance genes could be transferred from resistant to susceptible plant species to control important crop plant diseases. The recently cloned N gene of tobacco confers resistance to the viral pathogen, tobacco mosaic virus. We generated transgenic tomato plants bearing the N gene and demonstrate that N confers a hypersensitive response and effectively localizes tobacco mosaic virus to sites of inoculation in transgenic tomato, as it does in tobacco. The ability to reconstruct the N-mediated resistance response to tobacco mosaic virus in tomato demonstrates the utility of using isolated resistance genes to protect crop plants from diseases, and it demonstrates that all the components necessary for N-mediated resistance are conserved in tomato. PMID- 8710949 TI - CAX1, an H+/Ca2+ antiporter from Arabidopsis. AB - Reestablishment of the resting state after stimulus-coupled elevations of cytosolic-free Ca2+ requires the rapid removal of Ca2+ from the cytosol of plant cells. Here we describe the isolation of two genes, CAX1 and CAX2, from Arabidopsis thaliana that suppress a mutant of Saccharomyces cerevisiae that has a defect in vacuolar Ca2+ accumulation. Both genes encode polypeptides showing sequence similarities to microbial H+/Ca2+ antiporters. Experiments on vacuolar membrane-enriched vesicles isolated from yeast expressing CAX1 or CAX2 demonstrate that these genes encode high efficiency and low efficiency H+/Ca2+ exchangers, respectively. The properties of the CAX1 gene product indicate that it is the high capacity transporter responsible for maintaining low cytosolic free Ca2+ concentrations in plant cells by catalyzing pH gradient-energized vacuolar Ca2+ accumulation. PMID- 8710950 TI - Environmental and developmental signals modulate proline homeostasis: evidence for a negative transcriptional regulator. AB - In many plants, osmotic stress induces a rapid accumulation of proline through de novo synthesis from glutamate. This response is thought to play a pivotal role in osmotic stress tolerance [Kishor, P. B. K., Hong, Z., Miao, G.-H., Hu, C.-A. A. and Verma, D. P. S. (1995) Plant Physiol. 108, 1387-1394]. During recovery from osmotic stress, accumulated proline is rapidly oxidized to glutamate and the first step of this process is catalyzed by proline oxidase. We have isolated a full-length cDNA from Arabidopsis thaliana, At-POX, which maps to a single locus on chromosome 3 and that encodes a predicted polypeptide of 499 amino acids showing significant similarity with proline oxidase sequences from Drosophila and Saccharomyces cerevisiae (55.5% and 45.1%, respectively). The predicted location of the encoded polypeptide is the inner mitochondrial membrane. RNA gel blot analysis revealed that At-POX mRNA levels declined rapidly upon osmotic stress and this decline preceded proline accumulation. On the other hand, At-POX mRNA levels rapidly increased during recovery. Free proline, exogenously added to plants, was found to be an effective inducer of At-POX expression; indeed, At-POX was highly expressed in flowers and mature seeds where the proline level is higher relative to other organs of Arabidopsis. Our results indicate that stress- and developmentally derived signals interact to determine proline homeostasis in Arabidopsis. PMID- 8710951 TI - Patterns of kinship in groups of free-living sperm whales (Physeter macrocephalus) revealed by multiple molecular genetic analyses. AB - Mature female sperm whales (Physeter macrocephalus) live in socially cohesive groups of 10-30, which include immature animals of both sexes, and within which there is communal care of the young. We examined kinship in such groups using analyses of microsatellite DNA, mitochondrial DNA sequence, and sex-linked markers on samples of sloughed skin collected noninvasively from animals in three groups off the coast of Ecuador. Social groups were defined through photographic identification of individuals. Each group contained about 26 members, mostly female (79%). Relatedness was greater within groups, as compared to between groups. Particular mitochondrial haplotypes were characteristic of groups, but all groups contained more than one haplotype. The data are generally consistent with each group being comprised of several matrillines from which males disperse at about the age of 6 years. There are indications of paternal relatedness among grouped individuals with different mitochondrial haplotypes, suggesting long-term associations between different matrilines. PMID- 8710952 TI - A new physiologically approached in vitro test for quick evaluation of the hemolytic activity of surfactants. AB - New physiologically approached in vitro assays are presented which allow rather quick and uncomplicated determinations of the hemolytic activity of surfactants, intended mainly to solubilize poorly soluble drugs in parenteral dosage forms. The extent and the rates of the hemoglobin release from erythrocytes during the incubation of blood with increasing additions of surfactants is thought to be a very suitable and realistic estimation of the membrane damaging side effects of amphiphilic substances. In this study the extent of free hemoglobin released into the plasma is determined by a spectrophotometric method, as well as by an enzymatic determination measuring the lactate dehydrogenase (LDH) activity. LDH is released in a similar way from damaged erythrocytes. The results of these two methods are compared. They correspond satisfactorily. PMID- 8710953 TI - Formulation of controlled release drug preparations with antacid effect. AB - Opportunities for the formulation of long-acting antacid preparations were demonstrated summarizing the results of pharmaceutical technological experiments. The telemetric intragastric data were in correlation with the in vitro measurements and demonstrated the significantly higher bioavailability of long acting antacid preparations. PMID- 8710954 TI - Estimation of the gastric residence time of magnetic dosage forms using the Heidelberg capsule. AB - A new method for determining the gastrointestinal transit of magnetic dosage forms under the influence of an extracorporal magnet has been developed. A magnetic model dosage form was constructed consisting of small magnets attached to a pH-telemetering capsule (Heidelberg capsule). The transmission of the pH measurements was not affected by the magnetic field. The force of attraction of the magnetic model dosage form by the external magnet was characterised. In an in vivo study in five subjects the transmitted pH values showed indirectly that the gastric residence time of the magnetic model dosage form was prolonged significantly in the presence of an extracorporal magnet (t-Test, paired data, p < 0.05). PMID- 8710955 TI - Lymphatic transfer of macromolecules after intrapulmonary administration in the presence or absence of various absorption enhancers in rats. AB - We have developed a new method to evaluate the transfer of macromolecular drugs to intrapulmonary lymph nodes in rats after intrapulmonary administration. The transfer of fluorescein isothiocyanate dextrans (FDs) to the intrapulmonary lymph nodes was markedly higher than that to iliac lymph node, a non-intrapulmonary lymph node. The transfer of FDs to the intrapulmonary lymph nodes increased with increasing their molecular weights and the threshold for a high lymph node-to blood level ratio (CLN/CP) was between 10 and 20 kDa. The effects of absorption enhancers on the transfer of FDs to intrapulmonary lymph nodes were also examined in rats. Absorption enhancers used in this study were EDTA, sodium glycocholate (Na-GC), mixed micelles (MM), N-lauryl-beta-D-maltopyranoside (LM), sodium caprate (Na-Cap). The transfer of FD-20, FD-40 and FD-70 to intrapulmonary lymph nodes after intrapulmonary administration increased in the presence of LM. In particular, the lymphatic transfer of FD-40 was remarkably increased in the presence of LM. Similarly, Na-GC and MM improved the transfer of FD-40 to intrapulmonary lymph nodes. These results suggest that absorption enhancers such as LM, Na-GC and MM are effective for improving the transfer of macromolecular drugs to intrapulmonary lymph nodes. PMID- 8710956 TI - Three-dimensional solubility parameters and their use in characterising the permeation of drugs through the skin. AB - The physico-chemical properties of drug substances are major determinants of their transdermal absorption. In the present study the concept of the three dimensional solubility parameters of Hansen was applied in conjunction with the Bagley projection to describe the permeation of drugs and model substances through the skin. Drug permeation data from the literature were compared with the calculated solubility parameters of the drugs. It was demonstrated that the permeation of drugs can be estimated by their position in the Bagley diagram. There is a linear correlation between the logarithm of the skin permeation of drugs and the exchange cohesive energy for the steroids testosterone, progesterone, hydrocortisone acetate, corticosterone, cortisone, and dexamethasone. A linear correlation can be confirmed for the permeation of glyceryl trinitrate, digitoxin, oestradiol, scopolamine, atropine, diethylcarbamazine, fentanyl, and chlorpheniramine. In the case of morphine, codeine, sufentanil, meperidine and hydromorphone there is a linear relationship, too. PMID- 8710957 TI - [Phonophoretic permeation of procaine hydrochloride through an MDCK cell monolayer]. AB - With the aid of permanent cultured MDCK (Madin Darby Canine Kidney) epithelial cell a model for investigations dealing with phonophoresis effects was developed. The permeation of procaine hydrochloride through cell monolayers was examined while applying therapeutical ultrasound simultaneously. It could be shown that this permeation follows Higuchi kinetics. A comparison of the velocity factors, using a continuous irradiation of 1.0 W/cm2, shows about a 4.8 fold increase. Single Ultrasound pulses, however, result in a short-time enhancement of the permeation. The conclusion can be drawn that extent and velocity of the permeation of the permeation of procaine hydrochloride through MDCK monolayer can be controlled by phonophoresis. PMID- 8710958 TI - Use of a tube reservoir system for the pulsed release of nitroglycerin from transdermal plasters. AB - A novel tube-shaped drug reservoir system has been developed to enable the release of drugs from transdermal plasters to be actively controlled. The system provides a pulsed release of nitroglycerin from a thin PTFE tube, 22 cm long, two sections of which are filled with a solution of nitroglycerin in neutral oil. An electrolysis cell controls the continuous emptying of the tube, which results in the release of 0.5 mg doses of nitroglycerin at an interval of approximately 3 h. The tube reservoir and the electronic control can be incorporated into a normalsized transdermal plaster. PMID- 8710959 TI - Establishing the world list of schools of pharmacy. AB - The development of a world-wide list of schools of pharmacy by the former secretary of the Academic Section of the International Pharmacy Federation (F.I.P.) is described. Four print-based editions have been published since the first "preliminary" edition was made available to F.I.P. members in 1986. In February 1995, a version was launched on the World Wide Web, which has considerably facilitated the process of maintenance of the list through direct e mail contact of academics around the world with the editor. Links are provided to all schools which have established their own home-pages on the Web. The extent to which this resource will aid international communication between pharmacy academics is yet to be fully realised. PMID- 8710960 TI - Objective versus subjective: kudzu terminology. PMID- 8710961 TI - Regaining functional independence in the acute care setting following hip fracture. AB - BACKGROUND AND PURPOSE: Factors that predict functional recovery in the first few days following hip fracture and that may facilitate discharge to the home directly from the acute care setting have not been identified. This study investigated the attainment of key functional milestones by patients and discharge status from an acute care hospital following hip fracture. SUBJECTS: Subjects were 162 community-based individuals (59 men, 103 women) aged 60 years or older who were admitted to an acute care hospital following unilateral hip fracture. METHODS: Data on personal, medical surgical, hospital course, and acute rehabilitation factors as well as functional status and placement at the time of discharge were collected. Adjusted odds ratios were calculated to determine predictors of independence in seven types of transfers and ambulation activities and discharge directly to the home. RESULTS: Subjects who ambulated independently prior to fracture, stayed longer in the acute care setting, and received physical therapy on average more than once a day had improved odds of regaining independence in bed mobility, transfers, and ambulation. Subjects who regained independence and received physical therapy on average more than once a day had improved odds of discharge directly to the home from the acute care setting. Increasing age and postoperative complications reduced the odds of discharge directly home. CONCLUSION AND DISCUSSION: A substantial proportion of patients with hip fracture achieve independence in bed mobility and transfers and in ambulation with a walker during the early postoperative phase, although few progress to a higher level during a short-term stay in the acute care setting. Frequency of physical therapy, among other factors, appears to improve the odds of regaining functional independence and discharge directly to the home from the acute care setting. PMID- 8710962 TI - Work-related musculoskeletal disorders among physical therapists. AB - BACKGROUND AND PURPOSE: Increasing evidence suggests that musculoskeletal disorders are common in workers in the United States health care industry. Physical therapists, who commonly treat patients with these disorders, are also at risk for work-related musculoskeletal disorders (WMD) in the upper limbs and low back. The purpose of this study was to determine the prevalence of WMD during a 12-month period and the job factors that may be associated with these disorders in physical therapists. SUBJECTS: A four-page questionnaire was mailed to physical therapists (N = 1,160) who attended The University of Iowa between 1943 and 1993. Nine hundred twenty-eight questionnaires were returned (80% response rate) from physical therapists in 46 states. METHODS: Based on a literature review and pilot study of physical therapists, a survey instrument was constructed consisting of a symptom survey, a job-factor survey, and various demographic information. RESULTS: The highest prevalences of WMD among physical therapists were in the following anatomical areas: low back (45%), wrist/hand (29.6%), upper back (28.7%), and neck (24.7%). The job factor rated most likely to contribute to job-related musculoskeletal disorders was "lifting or transferring dependent patients." The prevalence of WMD in physical therapists also was affected by work setting, practice specialty, age of patient, and gender of therapist. CONCLUSION AND DISCUSSION: Specific strategies should be developed to reduce WMD in the practice of physical therapy. PMID- 8710963 TI - The effect of hamstring muscle stretching on standing posture and on lumbar and hip motions during forward bending. AB - BACKGROUND AND PURPOSE: The purpose of this study was to determine whether stretching the hamstring muscles affects extensibility, as indicated by straight leg raising (SLR), lumbopelvic posture, and the relative amounts of lumbar and hip motion during forward bending. SUBJECTS: Thirty-nine subjects without known musculoskeletal impairments of their spine or lower extremities and with tight hamstring muscles (ie, SLR of < or = 70 degrees) were randomly assigned to either a stretching group or a control group. METHODS: A three-dimensional digitizer was used to measure lumbar, pelvic, and hip positions while subjects were in a standing position and during partial and full forward bending. RESULTS: Straight leg raising and hip motion during late and total forward bending were increased after stretching. No changes occurred in standing posture or lumbar motion during forward bending. CONCLUSION AND DISCUSSION: The results of this study suggest that there is no relationship between hamstring muscle length and lumbopelvic posture. There was some indication, however, that stretching the hamstring muscles may affect motion during forward bending. PMID- 8710964 TI - Assessment of hamstring muscle length in school-aged children using the sit-and reach test and the inclinometer measure of hip joint angle. AB - BACKGROUND AND PURPOSE: The sit-and-reach test (SRT) is commonly used to assess flexibility of the spine and length of the hamstring muscles. The purposes of this study were (1) to describe hamstring muscle length as reflected by use of the SRT and the hip joint angle (HJA) in children, (2) to examine the correlation between SRT and HJA measurements, and (3) to examine gender differences for both measures. SUBJECTS: The participants were 410 school-aged children (211 girls, 199 boys). METHODS: Each child performed the SRT. In the final position, the SRT score was obtained and the HJA was measured using an inclinometer placed over the sacrum. RESULTS: A mean SRT value of 24 cm and a mean HJA value of 81 degrees were obtained for all subjects. There was a strong correlation between the SRT and HJA measurements (r = .76). There was a difference between boys and girls for both measures. CONCLUSION AND DISCUSSION: The results suggest differences in expectations for hamstring muscle length in boys and girls. Although scores for the SRT and HJA were correlated, we prefer to assess hamstring muscle length using HJA scores because these scores are not influenced by anthropometric factors or spinal mobility. The results of this study suggest that HJA measurements guide treatment more effectively than do SRT measurements. PMID- 8710965 TI - Conceptual learning processes in physical therapy students. AB - BACKGROUND AND PURPOSE: The purpose of this study was to investigate processes used by students in developing conceptual knowledge in physical therapy. SUBJECTS: The subjects were 10 first-year physical therapist students enrolled in a professional Master of Physical Therapy degree program. METHODS: Qualitative methods were used to collect data during a 15-week kinesiology course. Data were collected using open-ended interviews, observation, and journals kept by the students throughout the course. RESULTS: Several major themes emerged, including use of discussion, use of visualization, and use of experience by the students as they learned concepts in kinesiology. CONCLUSION AND DISCUSSION: The type of learning processes used by students in this study may be enhanced by educational methods such as collaboration and group learning, situated cognition and use of authentic contexts, cognitive apprenticeship, and whole-part-whole learning. Further research is needed to explore the relationship between student learning processes and teaching methods used in physical therapy education. PMID- 8710966 TI - The influence of subject and test design on dynamometric measurements of extremity muscles. AB - In the context of broader discussions of clinical dynamometry, earlier reviews have raised concerns about the potential effects of variations in subject factors and test procedures on measurements. None, however, have dealt exclusively with these effects. We therefore reviewed more than 200 articles to evaluate in detail the effects of variations in subject factors and test procedures on measurements. Factors relating to subjects that affected measurements were age, gender, weight, athletic background, disability, and limb dominance. Test conditions that led to variations in measurements were range of movement in which values were obtained, type of contraction or movement (concentric, eccentric, isokinetic, isometric, isotonic), pretest procedures (warm-up and gravity-correction procedures, starting position, stabilization, axes alignment, lever arm length, preload, damp/ramp settings), test conditions (speed, test sequence, rest intervals, feedback), and type of data analysis (the data selected and how they are manipulated). In the majority of the publications, the authors failed to provide sufficient detail for accurate replication of test procedures or for comparison with other studies. We advocate that the factors identified in this review be included whenever measurements obtained with a dynamometer are reported. Effective development of normative data, formation of ratios, comparison of measurements across studies, and relating measurements with other performance criteria (eg, measurements of functional performance) all require descriptions of variables relating to subjects and testing. Similarly, meaningful use of these measurements in clinical practice requires consideration and documentation of these variables. PMID- 8710967 TI - The Denny-Brown collection: useful resource for physical therapy practitioners and educators. PMID- 8710968 TI - Providing quality patient care in a reformed system: it can start by learning each others' names! PMID- 8710969 TI - Cutis laxa: etiology, pathophysiology, characteristics, and management. AB - Cutis laxa causes premature aging of the skin, which results in an abnormal appearance. Corrective, not aesthetic, surgery is one treatment option. A blepharoplasty and facelift would be two procedures that can be performed. PMID- 8710970 TI - Latex allergy: everyone is at risk. AB - Latex allergy often begins innocently with no symptoms or a slight case of contact dermatitis. As the allergy progresses, it becomes life-threatening. While this process puts our patients at great risk, we as health care workers fail to realize the risk we place our health and careers in, as we allow ourselves to become further exposed to the latex allergen at work and at home. PMID- 8710971 TI - A patient education guide for wired jaws. AB - Patients with wired jaws will be required to face a whole new lifestyle for up to 6 weeks following maxillofacial surgery. Changes in their lifestyles include relearning ways to care for their bodies, to eat their meals, and to communicate with family and health care professionals. Proper presurgical patient education is essential in order for these patients to gain confidence regarding self-care and for wounds to heal properly. PMID- 8710972 TI - Clinical photography simplified: developing a personal set of uniform views. AB - Clinical photography in plastic surgical settings often falls into the hands of staff nurses. Since physicians often don't have the luxury of employing full-time professional photographers, nurses and/or other auxiliary staff are responsible for shooting clinical photographs. A standard photographic routine, outlined in the following article, will ensure accurate photographic documentation. PMID- 8710973 TI - Facing challenges of managed care. PMID- 8710974 TI - Laser resurfacing. PMID- 8710976 TI - A research critique. Intraoperative music therapy: effects on anxiety, blood pressure. PMID- 8710975 TI - Ethical considerations for research on human subjects. PMID- 8710977 TI - Medications that may increase tendency to bleed. PMID- 8710978 TI - My child has hypospadias! AB - A child with a hypospadias anomaly needs a skilled surgeon to perform a delicate, complicated, sometimes staged, procedure successfully to prevent future psychological and physical problems. PMID- 8710979 TI - Rhytidectomy: from consultation to recovery. AB - Rhytidectomy, a popular cosmetic surgical procedure, requires a collaborative effort between the surgeon, nurse, and patient for a successful outcome. The prevention of complications is an ongoing process that begins with patient selection and continues through recovery. PMID- 8710980 TI - Improving the surgical screening process. AB - Computer and medical technologies have merged in health care practice and have clearly enhanced overall patient care. Nurses must possess computer literacy skills if they are to control their practice effectively in this high-tech environment. This article presents an overview of how one large midwestern hospital has enhanced patient care through the merging of computer technology with anesthesia and surgical pre-screening and preparation. PMID- 8710981 TI - Routine surgery, routine patients? Never! AB - The nursing process--assessing, diagnosing, planning, intervening, and evaluating -is a dynamic tool when properly used by the nurse. It ensures that nursing care is never routine. The following case study demonstrates how this process was used to guide the care of a patient with an unexpected corneal abrasion following meloplasty (chin augmentation) surgery. PMID- 8710982 TI - Anxiety of elective surgical patients' family members: relationship between anxiety levels, family characteristics. PMID- 8710983 TI - Preoperative and postoperative instructions for liposuction of jowl and chin. PMID- 8710984 TI - When the truth hurts. PMID- 8710985 TI - Readers should be fully briefed on adverse reactions of all glove products. PMID- 8710986 TI - Patient rights: is your center in compliance? PMID- 8710988 TI - What antibiotics have you taken recently? PMID- 8710987 TI - CPSN statistics offer confidence to nurses considering certification examination. PMID- 8710989 TI - President's message: compassion: a gift in action. PMID- 8710990 TI - [91st Annual meeting of Japanese Society of Psychiatry and Neurology. May 17-19, 1995. Nagasaki City, Japan. Abstracts]. PMID- 8710991 TI - [Social psychiatric study of the over-the-counter antitussive drug abuse; shifts resulting from modification of the formulae]. AB - We surveyed 22 patients (of whom 4 were women) with a history of antitussive drug abuse at a psychiatric hospital in the suburbs of Tokyo and at the Clinical Institute for Addiction Problems from 1 July 1986 to 30 June 1991. for changes in their pattern of abuse after the composition was altered. Here are our findings: 1) Researchers have long differed on whether ephedrine or codeine was responsible for the abuse of over-the-counter antitussive agents. The present study revealed that although codeine was still present in the new Bron syrup, its abuse was clearly on the decline in Japan, leading us therefore to conclude that psychological dependence on ephedrine was central to the spate of over-the counter antitussive abuse observed in the 1980s. Although we cannot ignore the mellow feeling codeine creates, it is difficult to give it more central role in the abuse of such drugs. 2) Users were divided into two groups, one that had used other agents before using antitussives, and another that had never used any other agent but antitussives. A comparison of these two groups revealed clear differences in learning behavior, social adaptation, familial environment and delinquent tendencies, prior to their drug abuse. 3) After the composition was altered, abuse spread to Tonin syrup manufactured by another laboratory, Bron tablets, new Bron syrup and ephedrine tablets. Relatively recently, there have been some new cases of abuse of Bron tablets and an increase in the number of cases who continued to use Bron syrup even after its composition was altered. The Medical literature also reveals that in many countries the abuse of both codeine and ephedrine is becoming a social problem, and clearly either agent may be abused separately. PMID- 8710992 TI - [Closing-in phenomena observed in patients with Alzheimer's disease--analysis of drawing behavior and eye movements]. AB - 'Closing-in' symptom is defined as a tendency to close-in on the model while performing constructive tasks. This unique symptom is one of the constructional apraxia often observed in Alzheimer's disease (AD), which is considered to be involved in visuospatial dysfunctions. However, little is known about the nature of the symptom and pathophysiology underlying the symptom. In this study, we examined drawing behavior, eye movements of AD (n = 19), multi-infarct dementia (n = 8) and age matched healthy subjects (n = 10). 1) In AD, contrary to age matched healthy subjects, the patients drew slowly with apparent difficulty and drawing behavior was disorganized. The copied figures were incomplete and often fragmented. The spatial relationships of the parts were lost each other during copying a Necker's cube. 2) The characteristic eye movements were observed in AD patients with apraxia, and closing-in symptom. These were consisted of gaze apraxia, and disorders of visual attention appeared similar to Balint's syndrome. 3) The eye movements of the AD patients with closing-in symptoms were classified into wandering of fixation, locking of fixation, and mixed types. The 'closing in' symptoms in copied figures were classified into three types, overlap, adherent, and near types. Then, the wandering type was seen both in near and adherent types, whereas the fixation type was only found in the overlap type. 4) SPECTs showed reduced RI activities in the parieto-occipital and in the frontal regions of AD patients with apraxia and closing-in symptoms. Our results suggest that AD patients may have a difficulty to create abstract space from concrete space. This may also imply regression to a primitive undifferentiated stage where gnostic and practic behavior is not differentiated in terms of space appreciation. PMID- 8710993 TI - [A nationwide investigation needs for rehabilitation of schizophrenic outpatients -the patients' attribute and actual conditions of living]. AB - In Japan we are very short of community resources for persons with mental disability. The authors, a board of resettlement, thought patients' actual conditions of living and needs for rehabilitation to let them live in the community had to be evaluated. Then in March 1993 the actual conditions of living and needs for rehabilitation of schizophrenic outpatients were investigated nationally. This article is the report on the patients' actual conditions of living. Investigation papers were sent to 358 institutions (286 hospitals and 72 clinics) which agreed to cooperate with the investigation. The 5186 investigation papers were received from 313 institutions. All of the papers except 18, which were lack of their ages, were analyzed. All of the schizophrenic outpatients, who consulted psychiatrists on one day during investigation, were considered objects of this research. Those who consented were included the research and psychiatrists filled in investigation papers. These institutions had a policy of intensive social resettlement activities and so on. Male patients were 55% and females were 45%. There were patients in the forties and females were older than males. Thirty-nine percent of them had been hospitalized once or twice. Thirty four percent of them had been hospitalized for less than one year. Eighteen percent of them had not been hospitalized. Fifty percent of patients answered there was no friend and acquaintance, and had a tendency to stand alone. Fifty three percent of patients lived with their parents, 21% with their spouses, and 17% alone. But 32% of females lived with their husbands. Sixteen percent worked for full-time jobs, 8% worked part-time jobs, 12% attended day care center 4% went to sheltered-workshops and only 1% went to rehabilitation-workshops for outpatients. While 13% didn't have a right to receive disability pension, the sources of income were job (30%), disability pension (30%), and welfare benefits (12%). Regarding the ability for living, both socially independent patients and daily independent were 36%. On the other hand, patients who needed support in daily life were 27%. This ability wasn't related to sex and age significantly (chi-square test). It is clear most of the patients were in isolation and depended on their families, so they needed support in income, accommodation and daily life. PMID- 8710994 TI - [Inhalant abusers and psychiatric symptoms]. AB - There are different opinions about the cause of chronic psychiatric symptoms observed in drug abusers between Japanese and foreign psychiatrists. The Japanese seem to recognize the chronic psychosis as the result of drug abuse. In the other hand, foreigners diagnose these cases as dual diagnosis of drug abuse and psychosis. Authors studied the problem in this research. One of the authors has examined 120 inhalant abusers of all, in- and out-patients in Kanagawa Prefectural Center of Psychiatry, Serigaya Hospital from 1991 to 1995. These patients were classified into three groups: psychosis group (23 patients), dependence group (51 patients) and abuse group (46 patients) according to their clinical courses and psychiatric symptoms. The psychosis group consists of patients who showed psychiatric symptoms such as hallucination, delusion and thought disturbance for long time after detoxification. The dependence group contains patients whose inhalant dependence was severe and met DSM-4 Diagnostic Criteria for Substance Dependence, but manifested no chronic psychiatric symptoms after detoxification. The patients belonging to abuse group were at the earlier stages of inhalant abuse and had no chronic psychiatric symptoms. The average age of the first inhalant abuse was 14.7 years old in the psychosis group, 14.8 years in the dependence group and 14.7 years in the abuse group. The average years of abuse was 9.0 years in the psychosis group, and 8.5 years in the dependence group. There was little difference between these two groups. The psychosis patients manifested chronic symptoms 5.7 years on average after the first abuse of inhalants. About one forth (26.1%) of the psychosis patients and only 5.9% of the dependence patients had family history of schizophrenia. The difference was statistically significant. These results suggest that chronic psychiatric symptoms are caused not only by inhalant abuse, but also by the genetic factors of psychosis of each patient. There have been several reports that many patients with dual diagnosis of substance dependence and other mental disorders are poly drug abusers. In our study, 43.4% of the psychosis group patients and 19.6% of the dependence group patients had the past history of abuse of other drugs including methamphetamine and marijuana. The difference was, however, not statistically significant. PMID- 8710995 TI - [The social factor that influences the psychological development of childhood and adolescence]. PMID- 8710996 TI - [Results of "questionnaire on the term and concept of schizophrenia" Committee on Concept and Terminology of Psychiatric Diseases. Sub-committee for Re-labelling the Term Schizophrenia in the Japanese Society of Psychiatry and Neurology]. PMID- 8710997 TI - [A research on the importance of the clinical certificate to the medical practitioners outside the field of psychiatry. Committee on graduate education sub-committee on Feasibility of Implementing Specialty Board Certification in the Japanese Society of Psychiatry and Neurology]. AB - OBJECTIVES AND METHODS: The main objectives of the research was to identify the merits and demerits of the specialty board certification system, that had been implemented in the other medical societies than the society of psychiatry in Japan. As a measure of the research, we conducted a survey by distributing questionnaires to medical institutions. The subjects of this survey included medical practitioners, other than the psychiatrists, who belong to general hospital, university hospital and the Japan Medical Association. FINDINGS: We received 228 responses out of the subjects. The findings were that 86% of the subjects have acquired the specialty board certificate, and that half of them have more than two kinds of qualifications on the average. Thus, the report suggests that the specialty board certification system have been long pervaded throughout the medical fields outside psychiatry. In addition, the subjects pointed out as the merits of the specialty board certification system that it helps to achieve the higher level of technique, and to organize and sophisticate as well as widen the knowledge of their specialty. They also reported that it contributes to instigate the life-long education and to earn identification and recognition of their specialty from other fields of medicine than their own. Fifty-three percent of the subjects acknowledged the necessity of the specialty board certification system while 13% regarded it unnecessary. This result proves that, despite higher percentage of the doctors with qualifications, the system itself is not approved of with full acknowledgement in the medical world. This rather negative attitudes toward the system is originated in the fact that acquiring and maintaining the certificate, i.e., renewal fees, are considerably high. In fact, as many as 62% of the subjects felt these points as disadvantages of the system. The medical practitioners outside the fields of psychiatry did not show any particular concern about the lack of the specialty board certification system in our field that is distinct as a field of medicine. Only 18% of the subjects expressed that urgent actualization of the system is desirable in our fields. In addition, securing the cites required for learning the technique and giving the exam to the applicants is the most important matter to be concerned. On the contrary, in other fields of medicine, the matter does not need to be discussed. Therefore, this is a problem unique to the fields of psychiatry. CONCLUSION: Although there are disadvantage of the specialty board certification system pointed out as shown above, we believe that it is necessary to introduce the system in the field of psychiatry. These disadvantage are mainly the matter of individual income. We need to consider the introduction of the system from the perspective of sophisticating the quality of psychiatry as a field of medicine in our country. We, the Subcommittee on feasibility of implementing specialty board certification in the Japanese Society of Psychiatry and Neurology have been contemplating over the system up to this moment. The progress has been made to the point of presenting, Discussion of the Specialty Board Certification System in Psychiatry, Yamauchi, May, 1994. PMID- 8710998 TI - [Huntington disease]. PMID- 8711000 TI - [Psychosocial problems in Huntington chorea. 1: Overview, project description. social and illness-related data]. AB - Huntington's Disease, a neuropsychiatric disorder with dominant inheritance and late manifestation is related to multiple psychosocial problems of the affected person, his life partner as well as their children (so-called individuals at risk). During a five years project the authors contacted 131 persons, directly and indirectly involved in problems with H.D. In the first part the authors present after a general survey sociodemographic and clinical data of the project. PMID- 8710999 TI - [The "neuroleptic threshold"--a review of the literature]. AB - The neuroleptic threshold theory raised the expectation that schizophrenic patients could be treated with an optimal, individual neuroleptic dosage, causing no side effects. Although most existing studies do not fulfill current methodological standards, a few methodologically convincing analyses confirm that good clinical results can be achieved with relatively low dosages (4 mg haloperidol equivalent per day). Thus, the neuroleptic threshold theory and methods related to it deserve further research. PMID- 8711001 TI - [Psychosocial problems in Huntington chorea. 2: Results and experiences from a 5 year project]. AB - The following article is informing about the experiences from a psychosocial project on Huntington's Disease (HD), a neuropsychiatric disorder with dominant inheritance. The project was running from July 1987 to October 1992 and was funded by the Robert-Bosch-Stiftung. During this project the authors contacted 78 involved families (131 persons), they offered psychotherapies, family-therapies, as well as meetings for patients and explored the subject by semi-structured interviews. The results of this project are presented in the following part. PMID- 8711002 TI - [Group psychotherapy in psychiatry--results of a pilot study and perspectives]. AB - This paper reports about first preliminary results of an investigation by DGPPN regarding group therapeutic work in psychiatric inpatient settings throughout the Federal Republic of Germany. The DGPPN sub-section "Group Therapy in Psychiatry" was set up by starting with this pilot study using a preliminary version of a questionnaire which is currently under revision for future investigations. This paper also deals with a brief historical survey of the origins of group work and group therapy in institutions. In regards to the installation of the new domain of a "Physician for Psychiatry and Psychotherapy" it is intended to theoretically consolidate group therapy in psychiatric facilities. The "Interpersonal Model" of Sullivan is being highlighted as a useful theory towards an integration of various theoretical perspectives such as biological, individual, interpersonal, and sociological ones. This integration is being stressed for education/ training and treatment aspects in psychiatry and psychotherapy as well. PMID- 8711003 TI - [Jealousy and jealous delusions as motive in homicide crimes--forensic and psychiatric aspects]. PMID- 8711004 TI - [Suicidal ideation in depressed patients with concomitant anxiety symptoms]. AB - Initial findings of panic disorder as an independent risk factor for suicidal ideation and behavior could not be replicated in studies with psychiatric patients. Instead, it was concluded that panic and anxiety disorders are risk factors when they co-occur with a primary mood disorder. In the present study, the effect of diagnostic comorbidity on rates of suicidality is analyzed on depressive inpatients treated at special depression wards. In a prospective follow-up study, suicidality and anxiety were assessed by means of a modified German version of the Diagnostic Interview Schedule (DIS). Patients with the symptom of panic attacks showed significantly elevated lifetime prevalence rates of suicidality in comparison with patients who did not report this additional symptom. For the follow-up period, however, there were no significant differences between these two groups. According to these results, the group of depressives with additional panic attacks is not more at risk for suicidal behavior, after being treated in an adequate manner. PMID- 8711005 TI - [Compulsory commitment to a psychiatric hospital--the status in Austria]. AB - Since 1991 in Austria a new law regulates involuntary admission at psychiatric hospitals and any other restriction of patient's freedom. Within 4 days a judge has to decide about further involuntary treatment. A new profession was established for involuntary patients support--the patients advocat. After a slow decrease of institutionalization rate in 1991 the number of involuntary admissions went up in 1993. The effects of the law upon psychiatric patients and treatment in psychiatric hospitals are discussed. Most psychiatrists (81%) are dissatisfied with the new legal situation because they see negative effects for some patients and have to spend a lot of time for juridical matters. PMID- 8711006 TI - [An unusual somatic symptom in a severe depressive episode]. PMID- 8711007 TI - [SSRI-induced akathesia]. PMID- 8711008 TI - [Life planning and psychosis--treatment of schizophrenia in unusual circumstances]. PMID- 8711009 TI - [A 20-year-old schizophrenic patient with acute leukemia]. PMID- 8711010 TI - [Sexual side-effects of clomipramine--a psychopharmacologic personal experience]. PMID- 8711011 TI - [Forensic practice between commonality, the law and psychiatry. 2]. PMID- 8711012 TI - Cognitive components of picture naming. AB - A substantial research literature documents the effects of diverse item attributes, task conditions, and participant characteristics on the case of picture naming. The authors review what the research has revealed about 3 generally accepted stages of naming a pictured object: object identification, name activation, and response generation. They also show that dual coding theory gives a coherent and plausible account of these findings without positing amodal conceptual representations, and they identify issues and methods that may further advance the understanding of picture naming and related cognitive tasks. PMID- 8711013 TI - Judgement research and neuropsychological assessment: a narrative review and meta analyses. AB - The authors review judgment research in the area of neuropsychological assessment. Topics include the reliability and validity of judgments, the appropriateness of confidence ratings, the value of training and experience, the cognitive processes of neuropsychologists, and the use of decision aids, including automated assessment programs and statistical prediction rules. Most of the research is on reliability and validity. The results indicate that neuropsychologists frequently make reliable and moderately valid judgments. However, for several tasks, experimenters have not studied the reliability and validity of judgments. PMID- 8711014 TI - Contingency detection and the contingent organization of behavior in interactions: implications for socioemotional development in infancy. AB - In this report, the authors review studies addressing the issue of contingencies in social and nonsocial contexts during infancy. The review is divided into 4 groups of studies that suggest that (a) young infants detect contingencies unrelated to their behavior; (b) infants detect contingencies involving their behavior; (c) the study of contingency is pertinent for addressing behavioral organization within parent-infant interaction; and (d) there is a link between behavioral contingency in early infancy and global measures of socioemotional development that are predictive of social functioning at later points during childhood. Throughout the report, the pertinence of infant emotional responses during contingency-related tasks is emphasized. Finally, the authors delineate certain enduring questions regarding contingency experience in infancy and suggest ways of organizing research to address some of them. PMID- 8711015 TI - The emotional Stroop task and psychopathology. AB - Attentional bias is a central feature of many cognitive theories of psychopathology. One of the most frequent methods of investigating such bias has been an emotional analog of the Stroop task. In this task, participants name the colors in which words are printed, and the words vary in their relevance to each theme of psychopathology. The authors review research showing that patients are often slower to name the color of a word associated with concerns relevant to their clinical condition. They address the causes and mechanisms underlying the phenomenon, focusing on J.D. Cohen, K. Dunbar, and J.L. McClelland's (1990) parallel distributed processing model. PMID- 8711016 TI - The sociocognitive model of dissociative identity disorder: a reexamination of the evidence. AB - According to the sociocognitive model of dissociative identity disorder (DID; formerly, multiple personality disorder), DID is not a valid psychiatric disorder of posttraumatic origin; rather, it is a creation of psychotherapy and the media. Support for the model was recently presented by N.P. Spanos (1994). In this article, the author reexamines the evidence for the model and concludes that it is based on numerous false assumptions about the psychopathology, assessment, and treatment of DID. Most recent research on the dissociative disorders does not support (and in fact disconfirms) the sociocognitive model, and many inferences drawn from previous research appear unwarranted. No reason exists to doubt the connection between DID and childhood trauma. Treatment recommendations that follow from the sociocognitive model may be harmful because they involve ignoring the posttraumatic symptomatology of persons with DID. PMID- 8711017 TI - Alcohol and aggression: a meta-analysis on the moderating effects of inhibitory cues, triggering events, and self-focused attention. AB - The authors conducted a meta-analysis of 49 studies to investigate 2 explanations of how alcohol increases aggression by decreasing sensitivity to cues that inhibit it. Both the level of anxiety and inhibition conflict moderated the difference between the aggressive behavior of sober and intoxicated participants, but neither level adequately accounted for variation in effect sizes. Additional analyses of 3 social psychological moderating variables-provocation, frustration, and self-focused attention-showed that the aggressiveness of intoxicated participants relative to sober ones increased as a function of frustration but decreased as a function of provocation and self-focused attention. The authors also examined the moderating effects of dose. PMID- 8711018 TI - Alcohol abuse in individuals exposed to trauma: a critical review. AB - In this article, the author critically reviews studies on the relationship between exposure to trauma, posttraumatic stress disorder (PTSD), and alcohol abuse. After establishing that strong relationships exist between exposure to traumatic events and alcohol problems, and particularly between the diagnoses of PTSD and alcoholism, the author discusses various factors, theories, and possible mechanisms to account for these associations. Moreover, she discusses applications of these findings to the assessment and treatment of people exposed to trauma who abuse alcohol. Finally, the author outlines novel methods for testing theoretical hypotheses and makes suggestions for methodological improvements in future research. PMID- 8711019 TI - Test-retest reliability of the Suicide Opinion Questionnaire. AB - Test-retest reliabilities for the Suicide Opinion Questionnaire are presented for eight samples across seven countries, representing both Eastern and Western cultures. Despite the variability of the samples and of the test-retest intervals, values fully support the reliability of the subscales. PMID- 8711020 TI - Relating depressive symptoms to Machiavellianism in a Turkish sample. AB - The Turkish forms of Beck Depression Inventory and The Mach IV Scale were administered to all 690 students of a Military Medical School. There was a significant correlation of .21 between scores on the Beck Depression Inventory and those on the Mach IV scale for the 658 men. This was not surprising, given the characteristics of Machiavellian people previously described. PMID- 8711021 TI - The Fagan Test of Infant Intelligence: predictive validity in a random sample. AB - The predictive validity of The Fagan Test of Infant Intelligence was investigated in a random sample of 100 boys and 96 girls from two Norwegian cities. Infants were assessed on the Fagan test between the ages of 7 to 9 months. A follow-up at 5 years included the Wechsler Preschool and Primary Scale of Intelligence-Revised (WPPSI-R) and three subtests from the Illinois Test of Psycholinguistic Abilities (ITPA). Low internal consistency of the Fagan test was found, with alphas of .10 and .02 for the two study sites. Fagan test-score correlated significantly with WPPSI-Verbal IQ and with scores on ITPA Auditory Reception for both samples, but the mean correlation was as low as .22. The prediction was not significantly higher for infants tested before 8 months of age than for those tested later. The study raises questions about use of the Fagan test as a screening tool in a nonrisk population. PMID- 8711022 TI - Suicide notes at symbolic ages. AB - Symbolic ages, like 40, 65, or 70, are often stocktaking occasions. Phillips and Smith in 1991 showed that suicides peak at such ages for married males and suggested that suicide notes may be an avenue to analyzing such stocktaking in people's lives. 40 notes from symbolic and nonsymbolic ages were compared based on a multidimensional clinical perspective. No differences were noted. Although caution is in order in accepting the null hypothesis, it is speculated that macrosocial phenomena may not be easily translatable into psychological ones. PMID- 8711023 TI - MMPI-2 profiles of men charged with murder or other offenses. AB - This study describes MMPI-2 profiles of men charged with murder or other offenses and examines how well the MMPI-2 differentiates between these groups. Records of 135 men charged with murder and 82 charged with other offenses were examined. Results do not support the use of the MMPI-2 to differentiate between murder and other defendants. Both groups may be described as having 6-8/8-6 profiles. The mean MMPI-2 profile for the murder group was compared to the MMPI profile described by Holcomb, Adams, Ponder, and Anderson in 1984. These mean profiles have generally the same configuration and scale elevations but show important differences. This suggests that further research needs to be conducted examining the psychometric equivalence of the MMPI and the MMPI-2. Also, caution should be used when making predictions based solely on MMPI-2 profiles. PMID- 8711024 TI - Risk screening for adverse outcomes in subacute care. AB - Purpose of this study was to identify variables near hospital admission that could identify patients at risk for placement, death, or readmission. The goals were to assess sensitivity and specificity of screening strategies in predicting adverse outcomes that deter or affect home health care. We evaluated whether generic screens might efficiently identify type of outcome. Criteria reported in the literature were used to predict outcomes for 1,332 admissions. Factors that discriminated type of outcome included comorbidity, mental status, living arrangement, transfer to special care, prior admission, iatrogenic trauma, and pending litigation. We conclude risk scores can accurately predict outcome of hospital treatment, which may be useful in targeting patients for intervention. Using billing data, although rather insensitive, was the most cost-effective strategy. PMID- 8711025 TI - Millon Clinical Multiaxial Inventory--II. Follow-up of patients with dissociative identity disorder. AB - This study is part of a two-year follow-up assessment of 35 out of 96 patients clinically diagnosed with Dissociative Identity Disorder, who were administered the Millon Clinical Multiaxial Inventory-II. Eight subjects achieved integration during the two-year follow-up period. Significant improvement was evident for raw scores on the Self-defeating, Borderline, Paranoid, Anxiety, Somatoform, Dysthymia, Alcohol Dependence, and Drug Dependent scales among both the integrated and nonintegrated patients, with clinically meaningful Base Rate reductions occurring on Self-defeating, Borderline, Avoidant, Passive-Aggressive, Anxiety, Dysthymia, and Major Depression scales following integration. Although Dissociative Identity Disorder presents with polysymptomatology, much treatment progress can be achieved during pre-integration and dramatic improvement follows integration. PMID- 8711026 TI - MMPI-2 items which correctly identified women with histories of childhood sexual abuse. AB - This study examined whether MMPI-2 items could be used to discriminate between 58 women who experienced childhood sexual abuse and 57 women who did not. A set of 48 items were identified which correctly classified 95% of the subjects. PMID- 8711027 TI - Cranial irradiation with Gaalas laser leads to naloxone reversible analgesia in rats. AB - Laser irradiation of the rat cranium can produce analgesia. The present experiment investigated the mechanism of such action. 27 rats received all possible combinations of laser (0, 6.4, and 12J/cm2) and naloxone (0, 5, and 10 mg/kg) prior to a hot plate test. Laser (820 nm, KHz pulsing, Omega Laser Systems, London) was applied to the rats' skulls and hind paw lick latencies (in seconds) were recorded immediately, 30 min., and 24 hr. after the administration of treatment. When animals were tested immediately following laser irradiation at 12J/cm2 significant analgesia resulted. Treatment with naloxone at either dose antagonised this effect, but naloxone produced no significant hyperalgesia when given alone. This suggests that opioid peptide mechanisms mediate the analgesic action of low-intensity laser irradiation of the cranium. PMID- 8711028 TI - Continued validation of the Multidimensional Perfectionism Scale. AB - Scores on the Multidimensional Perfectionism Scale have been correlated with measures of obsessive-compulsive tendencies for women, so the validity of scores on this scale for 41 men was examined. Scores on the Perfectionism Scale were significantly correlated (.47-.03) with scores on the Maudsley Obsessive Compulsive Inventory. PMID- 8711029 TI - College maladjustment may be related to alexithymia constructs with weakness of ego strength. AB - The authors examined the associations of scores on a measure of alexithymia and related basic dimensions of personality on college maladjustment in a sample of 387 students. Scores on two alexithymia constructs (difficulty identifying feelings and difficulty describing feelings) were significantly and positively correlated with scores on college maladjustment. Partial correlations controlling for scores on ego strength eliminated significant differences apparent in the simple correlations. College maladjustment may be related to the two alexithymia constructs with weakness of ego strength. PMID- 8711030 TI - Eliminating inappropriate classroom behavior using a DRO schedule: a preliminary study. AB - The purpose of this study was to eliminate out-of-seat behavior of a 14-yr.-old boy with learning disabilities using a single-subject design. Based on functional analysis, a Differential Reinforcement of Other Behavior interval schedule was used to eliminate an inappropriate behavior. During baseline, an average of 15 out-of-seat behavior problems were recorded. After 4 wk. of intervention, the inappropriate out-of-seat behavior was eliminated and on-task behavior improved. PMID- 8711031 TI - Comparison of attitudes of teachers of physical and musical education toward inclusion of children with disabilities. AB - 56 elementary teachers of physical education and 54 teachers of elementary music education responded to the Teacher Integration Attitudes Questionnaire which assesses teachers' attitudes regarding inclusion of children with disabilities into regular education settings. Analysis indicated that children with emotional and behavioral disorders were perceived less favorably by teachers of music education and children with orthopedic handicaps were perceived less favorably by teachers of physical education. The type and severity of disability appeared to influence teachers' attitudes towards including children with disabilities in regular education classrooms. PMID- 8711032 TI - Relationship between personal prayer and school-related attitudes among 11-16 year-old girls. AB - A sample of 392 girls between the ages of 11 and 16 years attending a state maintained single-sex Catholic secondary school completed six semantic differential scales of attitudes toward school and toward lessons concerned with English, music, religion, mathematics, and sports, together with information about paternal employment and their personal practice of prayer. The relationship between personal prayer and attitude toward school after controlling for age and social class was positive. PMID- 8711033 TI - Eating disorder and manic-depressive tendencies. PMID- 8711034 TI - Precued shifting of attention between cognitive sets in Parkinson patients. AB - The precueing paradigm developed by Posner has been used to examine visuospatial shifting of attention. In the current study, we modified such a paradigm so that it could be studied in nonvisuospatial domains and its component processes of disengagement, movement, and engagement could be analyzed in a similar fashion to the visuospatial domains. 14 patients with Parkinson's disease and 14 normal controls matched for age, sex, handedness, and years of education served as subjects. The speed of shifting attention was measured using the cost and benefit analysis. Analyses showed an over-all slowness in reaction time of patients with Parkinson's disease compared to the control group but without a concomitant slowness to engage, shift, and disengage their attention. PMID- 8711035 TI - Cognitive and psychodynamic correlates of depressive symptomatology. AB - 31 depressed, 26 nondepressed participants completed measures reflecting putative cognitive and psychodynamic characteristics of self-reported depression. Of the 16 variables 9 discriminated the groups in the expected direction. Ten characteristics correlated significantly with scores on the Beck Depression Inventory, 4 reflecting negative automatic thinking, two of which (Negative Self concepts and Expectations, Low Self-esteem) were important predictors of severity of depression. PMID- 8711036 TI - Evaluating questions used to assess cognitive orientation for very old adults. AB - A number of standardized tools have been developed to assist in the diagnosis of suspected dementia in aged people. In this study, 9 orientation questions were administered to 186 elderly Koreans, aged 60 to 108 years. Analysis showed a significantly inverse relation (r = -.70) between their ages and their performance on the questionnaire. The finding raises questions about the suitability of using standard measures of orientation with very old or with uneducated persons. PMID- 8711037 TI - Comment on "Jewish affiliation in relation to suicide rates". PMID- 8711038 TI - Compliance with follow-up care among adolescent males with sexually transmitted diseases. AB - The aim of the study was to investigate behavioral and sociodemographic characteristics of a sample of 45 male adolescents with sexually transmitted diseases who attended family planning clinics. Low rates of compliance with follow-up care were noted. Only 13 (28.9%) males returned to the clinic for their scheduled appointments. Neither behavioral nor sociodemographic variables were related to compliance. PMID- 8711039 TI - Perceived problem-solving and vocational identity: implication for nontraditional premedical students. AB - The purpose of this study was to examine the relationship between vocational identity and self-appraised problem solving. A total of 86 students enrolled in a special program took the Vocational Identity subscale of My Vocational Situation and the Problem Solving Inventory one week apart. Pearson product-moment correlations indicated a relationship between personal problem-solving appraisal and vocational identity, suggesting the inventories assess one construct. Implications for intervention strategies for the present population are discussed. PMID- 8711040 TI - Nonsmoking college students' attitudes toward smokers and smoking. AB - Although teenagers may begin smoking to enhance their social image, there is little evidence about how nonsmokers feel about their smoking peers or about nonsmokers' attitudes toward exposure to environmental tobacco smoke, so a 22 item questionnaire was administered to 547 nonsmoking college students. They reported that they were less likely to want a smoker for a roommate, date, or potential spouse. Most nonsmokers were bothered a great deal by environmental cigarette smoke and would want a smoke-free residence if living with a smoker. Almost all nonsmokers felt that environmental tobacco smoke increases the risk of serious disease. 92% strongly supported a ban on smoking in university classroom buildings and increased taxes on cigarettes. PMID- 8711041 TI - Fear of success among business students. AB - The concept of "Fear of Success" was measured with 352 male and female business students using the prompt, After first term finals, Ann(John) finds her(him)self at the top of her(his) Medical/Nursing school class. Analysis indicated a greater frequency of fear-of-success imagery among men than women and in particular to the John in Medical school and Ann in Nursing school cues. In addition, the Ann cue and the Medical school cue generated more fear-of-success responses among men than women. PMID- 8711042 TI - Adaptation strategies and transient niches of one middle-class inmate in prison. PMID- 8711043 TI - Revisiting fee policies. AB - Charging for missed sessions is presented as part of an ethical flexible fee policy that is being challenged and will require continued appraisal. PMID- 8711044 TI - An exploratory study of differences in developmental concerns of middle-aged men and women in India. AB - Responses of 56 (32 women, 24 men) subjects to the Men's Adult Life Experiences Inventory, a 97-item self-report assessment of the frequency and intensity of 11 developmentally related domains (relationships with parent, spouse, children, friends, death, health, job, leisure, sex, self-reflection, and pressure of time) of concerns of middle-aged men and women in India were similar. The men and women differed only in their response to the self-reflection and health domains of the Men's Adult Life Experiences Inventory. This study indicates that there is no midlife crisis in India. PMID- 8711045 TI - Psychiatric residents' scores on Machiavellianism. PMID- 8711046 TI - Casino use by college students. AB - College students' gambling behavior (N = 238) at the Foxwoods Resort Casino in Mashantucket, Connecticut was investigated using the South Oaks Gambling Screen. Of the 730 randomly sampled students, 238 returned usable questionnaires, a return rate of 32.5%. Only 35 (14.8%) of the students indicated having visited Foxwoods, of these, 26 (11% of the total) reported having gambled. Rates of problem were relatively low (6 students; 2.5%) and few underage students (3 students; 1.9%) reported using Foxwoods in the fall of 1994. As in other studies of casino gambling by students, slot machines and blackjack were the most frequent activities. PMID- 8711047 TI - Chronic psychiatric patients' use of caffeine: pharmacological effects and mechanisms. AB - The uses and effects of caffeine as a psychoactive drug in chronic psychiatric inpatient groups are described. Caffeine use and abuse is linked etiologically to diverse psychiatric disorders; its mechanisms of action are examined in relation to anxiety, anxiety neuroses, psychosis, schizophrenia, and caffeine intoxication and dependence. It is postulated that deleterious effects may result from the interaction of caffeine with commonly prescribed psychotropic drugs. A possible model of caffeine abuse is discussed. Increased public education about potential health problems related to caffeine consumption is suggested, and further controls of caffeine in psychiatric settings are recommended. PMID- 8711048 TI - Charging for missed sessions: ethical problem or straw person? AB - The subject of psychotherapists' charging patients for missed sessions is one of the important issues in professional ethics. It is better when not done at all, but the so-called flexible arrangements, although problematic in themselves, have less potential to damage the patient-therapist relationship than the exceptionless charging policy. Formal study is clearly needed. PMID- 8711049 TI - Psychology of computer use: XXXV. Differences in computer users' stress and self concept in college personnel and students. AB - 104 college students and 88 college faculty/staff at a midwestern university completed a questionnaire composed of the Computer Hassles Scale, a measure of computer users' stress, somatization/anxiety items from the Symptoms Checklist 90, a measure of stress reactions, and the Revised Personal Attribute Inventory, a measure of self-concept. Correlations indicated that for students there was a significant negative relationship between computer users' stress and self-concept (r = -.30), while for faculty/staff there was a significant positive relationship (r = .28). Regression analyses showed that self-concept moderated the relationship between computer users' stress and stress outcomes for only the faculty-staff sample. The moderator effect was interpreted using Linville's 1987 "buffering hypothesis," which suggests that persons with higher scores on self concept are less prone to experience stress-related outcomes like somatization/anxiety symptoms. PMID- 8711050 TI - Nurses' personality types based on the Myers-Briggs Type Indicator. PMID- 8711051 TI - Relationships between type A behavior, narcissism, and maternal closeness for college students in Japan, the United States of America, and the People's Republic of China. AB - The authors examined the relationships between Type A behavior and narcissism based on scores of college students in Japan, the United States of America, and the People's Republic of China. The scores on narcissism and Type A behavior differed significantly across the groups, being highest among the Chinese. In all three groups, the Type A scores were significantly and positively correlated with the scores on narcissism, and the latter were significantly and negatively correlated with the scores of mother's care. We refer in this study to cross cultural comparisons from viewpoints of sociocultural and psychological family structure. PMID- 8711053 TI - Learned helplessness in Drosophila melanogaster? PMID- 8711052 TI - Effects of a group rational-emotive behavior therapy program on the Type A behavior pattern. AB - A sample of 44 male Type A insurance representatives, selected by means of the Videotaped Structured Interview, were randomly assigned to a treatment (n = 22) and a delayed treatment control group (n = 22). The treatment group participated in 9 weekly sessions of group Rational-Emotive Behavior Therapy and were followed up after 10 weeks. After the control period, the delayed treatment control group received the same treatment program. Repeated measurements were obtained by means of the Videotaped Structured Interview, Jenkins Activity Survey, Cook-Medley Hostility Scale, and Type A Cognitive Questionnaire. Self and spouse/friend ratings of Type A behavior were obtained by means of the Bortner Rating Scale. Analysis indicated that, compared to the control condition, the therapy significantly reduced the intensity of Type A behavior and its time urgency component. These improvements were maintained at follow-up and were accompanied by self-reports of significant positive changes in Type A behavior and irrational beliefs. PMID- 8711054 TI - Factorial structure of the Arabic Children's Depression Inventory among Kuwaiti subjects. AB - The Arabic Children's Depression Inventory developed by Abdel-Khalek was administered to 1981 Kuwaiti middle-school students (995 boys and 986 girls). Their ages ranged from 10 to 16 years. Five orthogonal (varimax) factors were extracted. They were labeled generic depression, pessimism, feeling happy, weak concentration, and lack of loneliness. It was concluded that these interpretable factors pertained to the phenomenon of childhood depression, so the inventory can be recommended for use with Kuwaiti children and adolescents. PMID- 8711055 TI - Psychology of computer use: XXXVII. Computer-related stress and amount of computer experience. AB - 57 college students were surveyed using Hudiburg's revised Computer Technology Hassles Scale. From additional information, groups with various amounts of experience were formed. Comparison suggests that computer-related stress is not simply a by-product of increased interaction with computers. PMID- 8711056 TI - Social fears in hypochondriasis. AB - 152 students and inpatients of a psychosomatic clinic were divided into a hypochondriacal and a normal group with scores on the Hypochondriacal Beliefs and Disease Phobia scales from the Illness Attitude Scales. Social phobic characteristics were assessed by a clinical questionnaire. Significantly more subjects belonging to the hypochondriacal group scored above clinical cut-offs on measures of fear of criticism and fear of intimacy than subjects belonging to the normal group. Theoretical implications are discussed. PMID- 8711057 TI - Keep labels of variables simple and consistent: comment on the study of televised violence by Sutil, et al. PMID- 8711058 TI - Psychology of computer use: XXXIX. Prevalence of computer anxiety in British managers and professionals. AB - The prevalence of computer anxiety among British managers and professionals was investigated. Questionnaire data were obtained from a sample of 235 British managers and professionals. Prevalence was defined as the percentage of individuals in the sample with scores above the midpoint on the computer-anxiety scale. A prevalence rate of 21.3% was found. The prevalence among women was double that among men. This rate is considered very high and may have significant negative effects on productivity or ability to adapt to new computer-based technology. Further, the results imply that scales assessing constructs relevant to rapidly changing issues such as computers need to be constantly updated. PMID- 8711060 TI - Examination of drug-induced and isolation-induced disruptions of prepulse inhibition as models to screen antipsychotic drugs. AB - Prepulse inhibition (PPI) of an acoustic startle response is impaired in schizophrenics. PPI can also be studied in the rat, and is impaired by dopamine (DA) D2/3 receptor agonists such as apomorphine. This disruption is reversed by DA antagonists, leading to proposals that this approach may be a useful means to identify novel antipsychotics. There is also evidence to suggest a role of serotonergic (5-HT) and glutamatergic systems in schizophrenia, and accordingly PPI can be disrupted by the 5-HT2 agonist DOI, and the non-competitive NMDA antagonist, dizocilpine. In the present study we have examined the effect of four antipsychotic drugs, haloperidol (0.1-0.3 mg/kg), raclopride (0.03-0.3 mg/kg), risperidone (0.3-3 mg/kg) and clozapine (0.0001-10 mg/kg), against the PPI disruptions induced by apomorphine (0.5 mg/kg), DOI (3 mg/kg) and dizocilpine (0.15 mg/kg). Furthermore, these drugs have been examined for their ability to restore a PPI deficit produced by housing rats under conditions of social isolation. All drugs except clozapine reversed an apomorphine-induced disruption. However, clozapine and risperidone, but not raclopride and haloperidol, reversed a DOI-induced disruption. Only risperidone was effective in restoring a PPI deficit produced by dizocilpine. In contrast to the drug-induced disruptions which were differentially sensitive to the various neuroleptics, isolation induced disruptions were restored by each drug. These results support the idea that non-drug induced disruptions of PPI, such as social isolation, may be a more viable approach to identify novel antipsychotics. PMID- 8711059 TI - Behavioural consequences of maternal exposure to natural cannabinoids in rats. AB - Cannabis sativa preparations (hashish, marijuana) are the most widely used illicit drugs during pregnancy in Western countries. The possible long-term consequences for the child of in utero exposure to cannabis derivatives are still poorly understood. Animal models of perinatal cannabinoid exposure provide a useful tool for examining the developmental effects of cannabinoids. Behavioral consequences of maternal exposure to either cannabis preparations or to its main psychoactive component, delta 9-tetrahydrocannabinol (THC) in rat models are reviewed in this paper. Maternal exposure to cannabinoids resulted in alteration in the pattern of ontogeny of spontaneous locomotor and exploratory behavior in the offspring. Adult animals exposed during gestational and lactational periods exhibited persistent alterations in the behavioral response to novelty, social interactions, sexual orientation and sexual behavior. They also showed a lack of habituation and reactivity to different illumination conditions. Adult offspring of both sexes also displayed a characteristic increase in spontaneous and water induced grooming behavior. Some of the effects were dependent on the sex of the animals being studied, and the dose of cannabinoid administered to the mother during gestational and lactational periods. Maternal exposure to low doses of THC sensitized the adult offspring of both sexes to the reinforcing effects of morphine, as measured in a conditioned place preference paradigm. The existence of sexual dimorphisms on the developmental effects of cannabinoids, the role of sex steroids, glucocorticoids, and pituitary hormones, the possible participation of cortical projecting monoaminergic systems, and the mediation of the recently described cannabinoid receptors are also analyzed. The information obtained in animal studies is compared to the few data available on the long-term behavioral and cognitive effects on in utero exposure to cannabis in humans. PMID- 8711061 TI - Increased sensitivity to the sensorimotor gating-disruptive effects of apomorphine after lesions of medial prefrontal cortex or ventral hippocampus in adult rats. AB - Sensorimotor gating of the startle reflex is impaired in humans with schizophrenia and in rats after mesolimbic D2 dopamine receptor activation. The loss of startle gating after D2 activation in rats has been used as an animal model of impaired sensorimotor gating in schizophrenia, because the ability of antipsychotics to restore startle gating in D2-activated rats correlates significantly with antipsychotic clinical potency. Substantial evidence indicates that the pathophysiology of schizophrenia includes structural and functional deficits in prefrontal and temporal regions, particularly the dorsolateral prefrontal cortex and the hippocampus and parahippocampal gyrus. The present study assessed startle gating in adult rats after ibotenic acid lesions of the medial prefrontal cortex or ventral hippocampus. Medial prefrontal cortex lesioned rats exhibited normal startle amplitude and normal sensorimotor gating, as reflected by prepulse inhibition (PPI) of the startle reflex. Hippocampus lesioned rats exhibited elevated startle amplitude, and similar to rats with medial prefrontal cortex lesions, did not show significant changes in basal PPI. Low doses of the mixed dopamine agonist apomorphine did not significantly reduce PPI in sham lesioned rats, but significantly disrupted PPI in both medial prefrontal cortex- and ventral hippo-campus lesioned rats. These data are consistent with the hypothesis that cell damage in frontal and temporal cortex increases the sensitivity to the sensorimotor gating-disruptive effects of dopamine receptor activation. PMID- 8711062 TI - Neonatal excitotoxic hippocampal damage in rats causes post-pubertal changes in prepulse inhibition of startle and its disruption by apomorphine. AB - Neonatal excitotoxic hippocampal damage in the rat results in postpubertal onset of a variety of abnormal behaviors related to excessive dopaminergic transmission in the mesolimbic/nigrostriatal system, and thus may be considered an animal model of some aspects of schizophrenia. Because sensorimotor gating is impaired in adult patients with schizophrenia and in rats with experimentally induced mesolimbic dopamine hyperactivity, the present experiments investigated the effects of neonatal (postnatal day 7, PD7) ibotenic acid (3 micrograms) lesions of the ventral hippocampus (VH) on the amplitude and prepulse inhibition (PPI) of acoustic startle in prepubertal (PD35) and postpubertal (PD56) rats. Startle was elicited using 105 and 118-dB pulses alone or preceded by 4, 8, or 16 dB above background prepulses in rats treated with vehicle or apomorphine (APO; 0.025 or 0.1 mg/kg SC). At PD35, PPI in VH-lesioned rats did not differ significantly from these measures in sham operated rats. Apomorphine significantly increased startle amplitude and reduced PPI in both sham operated and VH-lesioned rats at PD35. At PD56, startle amplitude in VH-lesioned rats was not significantly different from controls, but PPI was reduced significantly compared to controls. Ventral hippocampus lesioned rats also exhibited an exaggerated reduction in PPI after treatment with APO. These findings provide further evidence of postpubertal impairments that may be related to increased mesolimbic dopamine transmission and receptor sensitivity in rats with neonatal hippocampal damage, and provide further support for the fidelity of this animal model of schizophrenia. PMID- 8711063 TI - Alterations in hippocampal hemicholinium-3 binding and related behavioural and biochemical changes after prenatal phenobarbitone exposure. AB - Previous studies demonstrated postsynaptic septohippocampal cholinergic alterations after early exposure to phenobarbital. The present study was designed to ascertain possible corresponding presynaptic alterations while confirming the known behavioral deficits and extending previous findings on postsynaptic cholinergic alterations. Pregnant heterogeneous mice received milled mouse food containing 3 g/kg phenobarbital on gestation days 9-18. At age 50 days, [3H]hemicholinium-3 binding, which labels the presynaptic transporter for high affinity choline uptake, was increased in treated mice by 100% (P < 0.001). This change was not accompanied by a change in the affinity of the transporter to the ligand. Another group of offspring was tested for hippocampus-related behaviors. Consistent with our previous studies in the Morris maze, treated animals took longer to reach the platform in the place test as compared to control, and swam fewer times over the missing platform location in the spatial probe test. In the eight-arm maze, the treated offspring needed more entries than control to visit all the arms. In the spontaneous alternation test, the treated mice showed fewer alternations than controls. Biochemically, as in our previous results, prenatal phenobarbital exposure resulted in an increase in the degree of stimulation of inositol phosphate formation by carbachol (P < 0.05), an action presumed to occur at postsynaptic muscarinic receptors. While the present results show that the effect of a combination of raised K+ in the presence of physostigmine and carbachol was significantly greater in barbiturate-treated mice (P < 0.05), the action of K+ in the presence of physostigmine, but without carbachol, was not affected by the phenobarbital treatment. The results point to the uniqueness of outcome of early insults where alterations along nerve conduction cascades do not necessarily follow the common rules in that upregulation could simultaneously occur both pre- and post synaptically. PMID- 8711064 TI - Effects of prenatal diazepam on two-way avoidance behavior, swimming navigation and brain levels of benzodiazepine-like molecules in male Roman high- and low avoidance rats. AB - Utilizing psychogenetically selected Roman high- and low-avoidance rats (RHA/Verh and RLA/Verh), the present experiments investigated the effects of prenatally administered vehicle and diazepam (1 and 3 mg/kg per day, SC) on the behavior and neurochemistry of adult, male offspring. Active, two-way avoidance behavior was analyzed in 96 rats, at 6 months of age, and swimming navigation in 68 others, at 11 months. Three weeks after testing, selected brain areas from the latter animals were immunoassayed for benzodiazepine (BZD)-like molecules. The 3 mg/kg dose of diazepam both decreased freezing behavior in the shuttle box and reduced the hippocampal content of BZD-like molecules in the RLA/Verh male rats. Swimming navigation (spatial learning), at which the RLA/Verh rats were more adept, was not specifically affected by prenatal diazepam in either rat line. The possibility exists that an increased hippocampal release of BZD-like substances may be necessary to alter shuttle box behavior in RLA/Verh rats. PMID- 8711065 TI - Limited changes of mouse maternal care after prenatal oxazepam: dissociation from pup-related stimulus perception. AB - There is evidence that activity at the GABA/benzodiazepine receptor complex in specific brain areas might be enhanced during rodent motherhood. We tested the hypothesis that the manipulation of this neurochemical system by prenatal benzodiazepine exposure affects typical behavioral responses of lactating mouse dams. Outbred CD-1 mouse fetuses were administered either oxazepam (OX, 15 mg/kg) or vehicle twice a day on embryonic days 12-16 and fostered at birth to untreated dams. Female offspring were subsequently mated at the young adult stage. In a first experiment, the behavioural repertoire of the two groups of lactating females was scored (single 10-min session) from postpartum days 3 to 18. When compared with VEH dams, OX females showed a shorter duration of pup-sniffing at 7 10 days and enhanced crouching behaviour when pups had reached the age of 14-18 days. In addition, OX-treated dams used more cotton for nest construction than the controls. The two female groups were differentiated only in the presence of their offspring. In a second experiment aimed at investigating possible OX induced changes in pup-stimulus perception, the same lactating females were challenged in sequence on postpartum day 8 with three different patterns of pup related cues consisting of: three 8-day-old live male pups (LP), three same-age dead pups DP, or three dead pups accompanied by pre-recorded ultrasounds (DPU). In the absence of carry-over effects of prenatal dam treatment, LP stimuli elicited a higher frequency of sniffing and digging than the others, whereas the level of licking, and grooming was reduced. In conclusion, the present results indicate that the slight alteration in maternal care resulting from prenatal OX treatment can be dissociated from changes in pup-related stimulus perception. PMID- 8711066 TI - Irreversible impairment of active avoidance behavior in rats prenatally exposed to mild concentrations of carbon monoxide. AB - Wistar female rats were exposed to relatively mild concentrations of carbon monoxide (75 and 150 ppm) from day 0 to day 20 of pregnancy. The results show that prenatal exposure to CO (150 ppm) significantly impairs the acquisition of a two-way active avoidance task in 3-month-old male rats as well as the acquisition and reacquisition of this schedule in 18-month-old animals subjected to six daily 20-trial sessions. These deficits do not seem to be attributable to alterations of a non-associative nature, as the intertrial activity and the escape response latencies in CO exposed animals were not significantly affected with respect to controls. These findings, showing that gestational exposure to CO induces in rat offspring permanent learning and memory impairment, confirm that the offspring of smoking mothers may be at considerably greater risk than current epidemiological studies on birthweight and neonatal mortality suggest. PMID- 8711067 TI - A comparison of behavioural effects of prenatally administered oxazepam in mice exposed to open-fields in the laboratory and the real world. AB - Prenatal benzodiazepine exposure has been reported to result in abnormal neurobehavioural development in laboratory animals but little is known about the behavioural relevance of this effect ina naturalistic environment. In this study, outbred CD-1 male mice were prenatally exposed to oxazepam (15 mg/kg per os, twice daily) on days 12-16 of fetal life and fostered at birth to untreated dams. At adulthood, each mouse was fitted with a radio collar and its first reactions assessed. Three hours later, behavioural and exploratory activities were recorded in a laboratory open field, and 24 h later in a natural setting. Immediate reactions to the radio collar were higher in the oxazepam-treated mice than in controls consisting of more attempts to remove it and an increase of push digging. The attempts to remove the collar were still evident in oxazepam treated mice tested in the laboratory open-field 3 h later. Moreover, oxazepam increased the frequency of grooming and reduced walking in both the laboratory and the natural settings. In the natural settings running was increased during the initial 30-min test, while a pronounced level of grooming and a lower frequency of eating were observed 140 min after release. Frequency of sniffing, grooming, and rearing behaviours were higher in the laboratory test when compared to the natural settings. On the other hand, prolonged bouts of sniffing were recorded in the natural environment. These findings permit separation of robust drug effects (increased grooming, reduced walking) from situation-dependent effects, the natural environment revealing, in addition, more subtle effects. PMID- 8711068 TI - Ontogenetic studies of tolerance development: effects of chronic morphine on the hypothalamic-pituitary-adrenal axis. AB - Endogenous opiates are important regulators of the hypothalamic-pituitary-adrenal (HPA) axis in rats. Tolerance clearly develops to morphine-induced stimulation of the HPA axis in adult rats (Ignar and Kuhn 1990). The goal of the present study was to determine whether tolerance to morphine-induced stimulation of the HPA axis developed in neonatal and weanling rats treated chronically with morphine. Rats were injected with morphine or saline between days 4-8 postnatal (pups) or days 21-25 (weanlings) and tolerance assessed by determining dose-response curves for ACTH and corticosterone secretion following an acute morphine challenge. Weanlings displayed marked tolerance to the stimulation of ACTH and corticosterone secretion by morphine. Tolerance was also observed in pups to morphine-stimulated ACTH and corticosterone release. These findings suggest that the relative adaptability of the HPA axis to chronic morphine in neonatal and weanling rats is similar. PMID- 8711070 TI - Further insights into transsexualism. AB - Transsexualism is a challenging entity posing diagnostical, etiological, ethical and moral questions with no general consensus of opinion and a controversial management policy. The present article tries to deal with these questions by drawing a parallel with another psychiatric syndrome in which there is also a disturbance in body image, namely anorexia nervosa. The parallel is at the level of clinical presentation, psychodynamic etiology and their common socioepidemic dimension. PMID- 8711069 TI - Independent mediation of unconditioned motor behavior by striatal D1 and D2 receptors in rats depleted of dopamine as neonates. AB - The effects of systemic administration of DA receptor antagonists suggest that unconditioned motor behavior in rats depleted of DA as neonates continues to be dependent upon dopaminergic transmission, yet the specific contribution of D1 and D2 receptors to these behaviors has been altered. The purpose of the present study was to determine whether these depletion-induced receptor changes are occurring at the level of striatal DA terminals and their targets. The ability of bilateral intrastriatal injections (0.5 microliter) of DA receptor antagonists to induce motoric deficits was determined in adult rats treated with vehicle or 6 OHDA (100 micrograms, intraventricular) on postnatal day 3. Administration of the D1-like antagonist SCH 23390 (0.5-2.0 micrograms) or the D2-like antagonist clebopride (1.0-4.0 micrograms) induced dose-dependent akinesia, catalepsy, and somatosensory neglect in vehicle-treated controls. In contrast, neither antagonist produced deficits in rats depleted of forebrain DA as neonates. However, combined administration of SCH 23390 + clebopride induced similar akinesia, catalepsy, and somatosensory neglect in both controls and DA depleted animals. Animals depleted of DA were more sensitive than controls to the low doses of this combined D1 + D2 antagonism. These results demonstrate that activation of striatal DA receptors remains necessary for unconditioned motor behavior in rats depleted of DA as neonates. However, the specific contributions of D1- and D2-like receptors to these behaviors differ between intact animals and those depleted of DA as neonates. The ability of endogenous DA acting at either D1 or D2 receptors to support spontaneous motor behavior in rats depleted of DA as neonates may contribute to their relative sparing from parkinsonian deficits. PMID- 8711071 TI - Gene-environment interaction in schizophrenia: season-of-birth effect reveals etiologically different subgroups. AB - Compared to the general population, one consistent finding in schizophrenia research is a significant surplus of schizophrenic births in the winter/spring months. There is little evidence that this is attributed to statistical artefacts. The "harmful effects' hypothesis offers the most plausible explanation for this phenomenon. Exogenous harmful effects, predominant during the cold season, may affect the developing immature fetal brain and thus constitute some of the factors predisposing to schizophrenic breakdown in adulthood. Neuropathological and epidemiological studies point to the second trimenon of gestation as the crucial period of fetal brain maturation. Recently, some studies found that the surplus in schizophrenic winter/spring births is mainly due to sporadic forms. In contrast, patients with high genetic risk of the disease even tended to have a birth deficit during this period. This suggests that schizophrenia is not a disease entity but consists of etiologically distinct subgroups on which the influence of genes and/or environment has to be weighted differently. On the one hand, in sporadic forms of the disease exogenous noxious agents may be of major etiological importance. On the other hand, in fetuses at high genetic risk neurodevelopment may already be disturbed due to a genetic defect and additional environmentally noxious agents can cause abortions, stillbirths and sudden infant deaths. PMID- 8711072 TI - The enigma of pseudohallucinations: current meanings and usage. AB - This study investigates the view of 2 samples of psychiatrists about pseudohallucinations (PH) and their perceived value in clinical practice. Among UK and Irish consultant psychiatrists, 87% expressed belief in PH, but 63% thought that the concept was confusing and only 58% thought PH were clinically useful. Recently appointed consultants were more positive about PH. One-third of respondents were sceptical: and even the more enthusiastic gave variable and inconsistent responses. Many psychiatrists do not value the concept of PH or do not use it. Those who do favour it use it inconsistently. The concept gives rise to much confusion, and is weak because it is defined negatively. It should be abandoned in favour of a more precise clinical approach to describing false perceptions. PMID- 8711073 TI - Lilliputian hallucinations in schizophrenia: case report and review of literature. AB - A patient with chronic schizophrenia who had visual hallucinations of lilliputian nature is described. The authors review the studies on prevalence of visual hallucinations in schizophrenic patients and suggest that visual hallucinations may be more common in schizophrenia than previously acknowledged. The authors suggest a routine enquiry about visual hallucination in clinical practice and emphasize the need for further research under controlled conditions. PMID- 8711074 TI - Patterns of use and changes in diagnosis during first admission. National Case Register Study. AB - The goal of this study was to describe patterns of diagnosis and to explore the extent to which diagnosis changes during first-in-life psychiatric admissions. All 2,998 first admissions to Israeli psychiatric wards in 1989 were studied. Diagnosis did not change in at least 60% of the cases. Diagnoses in order of stability were: mental retardation (84%), substance abuse (82%), organic conditions (77.5%), neurotic (75%), schizophrenia (74%), personality disorders (73%), affective (68%), childhood (55%), paranoid (45%) and V-codes (22%). There was less change in diagnosis for patients over 45 (37.5%), than for patients aged 19-44 (43.2%) and 15-18 (57.8%). Diagnoses assigned at admission to first hospitalization are not likely to change during that hospitalization. PMID- 8711075 TI - Factor analysis of the Frankfurt Complaint Questionnaire in a Spanish sample. AB - The Frankfurt Complaint Questionnaire (FCQ) is a self-assessment instrument designed to evaluate the subjective experiences (SE) of psychotic patients. It is composed of 98 yes-no items grouped in ten phenomenological subscales. The original validation of the FCQ included a factor analysis with a four-factor structure. No further studies of factor validity have been carried out. The present study aimed to replicate the factor structure of the FCQ in a Spanish sample. The sample was composed of 286 consecutively admitted patients due to a recrudescence of their psychotic symptoms. They were evaluated through a semistructured interview for schizophrenia and diagnosed according to DSM-III-R criteria. Organic mental disorders were excluded. Sixteen patients who failed to fulfill the FCQ were excluded. The sample finally comprised 270 patients. A factor structure of FCQ items comprising 25 factors was found. The first factor obtained the highest explained variance, and most items obtained their highest load on the first factor. These results strongly suggest a unidimensionality underlying FCQ items. A new scale composed of 18 items was derived from those with higher weights in the first factor. The new scale of SE was presented in a Likert format to demonstrate their frequency and intensity more clearly. PMID- 8711076 TI - Recognition of hallucinations: a new multidimensional model and methodology. AB - Both from the clinical and mathematical perspectives, symptom recognition has received less attention than disease recognition. To redress this balance, it is imperative that multidimensional models are constructed for each and all mental symptoms. This paper offers one such model for "hallucinations', and a set of prototypical data comparing the performance of pattern recognition techniques (cluster and discriminant analyses) and neural networks (Kohonen and backpropagation). It is concluded that multidimensional models are less wasteful of information than (current) categorial ones. Because of this and of the fact that symptom structure is likely to be "isomorphic' with the brain region where the corresponding signal is generated, it is recommended that multidimensional models are preferentially used in neurobiological research. PMID- 8711077 TI - Perceived parental bonding in anorectic and bulimic patients. AB - Psychodynamic clinical studies as well as empirical research have pointed out the importance and the specificity of object relations in eating disorders. Given this, the aim of this work was to identify the presence of significant differences in the object relations patterns, as described by the Parental Bonding Instrument (PBI), between anorectic and bulimic subjects. PBI was administered to 42 anorectic female and 26 bulimic female subjects, consecutively admitted to the Eating Disorders Outpatient Unit of H San Raffaele of Milan. Profile analysis revealed the presence of significant (T2 = 11.3453, d.f. 3,64, p = 0.017) qualitative differences in PBI profiles between anorectic and bulimic subjects. The most striking difference was represented by the bulimics' view of their parents as both caring and overwhelming, which contrasted with the anorectics' perception of their parents as absolutely caring. PMID- 8711078 TI - Word associative production in affective versus schizophrenic psychoses. AB - The production of association word to stimulus words, which was found to be correlated with conceptual disorganization, as clinically measured by the Brief Psychiatric Rating Scale, was developed as a quantifiable measure of formal thought disorder. Associative word production in patients with affective psychoses (acute episodes of mania or schizoaffective disorder) was found to be higher in a statistically significant manner than in patients with acute episode of paranoid schizophrenia. The production of associative words in the two groups of acutely psychotic patients was significantly higher than in normal subjects, unipolar depressed, or residual schizophrenic patients. These quantitative differences reflected qualitative differences in the pattern of the production of word associations. Indeed, while patients with paranoid schizophrenia showed a sinusoidal-like type of oscillation in associative word production, patients with affective psychoses were characterized by exponential-like phases in associative word production. Associative word production may thus serve as a simple quantitative test for differentiating formal thought disorder in acute psychoses between patients with mania and patients with schizophrenia. PMID- 8711079 TI - Can loss and grief activate latent neoplasia? A clinical case of possible interaction between genetic risk and stress in breast cancer. AB - BACKGROUND: Nineteenth-century physicians working with cancer patients frequently reported that severe emotional losses and grief occurred in some cases before cancer. Research on psychoneuroimmunology seems to reactivate this interest and could give a possible convincing background. METHOD: We recently observed the case of a 45-year-old woman who developed symptoms of breast cancer (ductal infiltrating carcinoma, T1, N1, M0) 3 years after the loss of her only child. The affection for the child compensated inadequacies of her marriage, which became increasingly unstable. The child died in a dramatic accident at home, while his mother was near him but unable to help. RESULTS: The patient had a moderate genetic-familial risk of mammary cancer. Animal studies strongly suggest that stress, through neuroimmunomodulatory mediatory mechanisms, can significantly affect appearance and progression of mammary cancer. The patient developed long lasting intense grief, despair and hopelessness. DISCUSSION: We discuss the case from three main standpoints: genetic and risk factors for breast cancer; findings which demonstrated a relationship between stress, neuroimmunomodulation and cancer in animals; data from autopsy studies which found foci of latent breast cancer in 20% of healthy women in the same age range. Stress does not create cancer. However we hypothesize that in this case it could have contributed by the activation of a latent neoplasia and/or by impairing immunosurveillance during a critical life phase. PMID- 8711081 TI - Seizures and the Dandy-Walker syndrome: a case of suspected pseudoseizures. AB - BACKGROUND: A major pitfall in the diagnosis of conversion disorder is the difficulty in ruling out organic causes. METHODS. A 19-year-old female was referred with seizures and the Dandy-Walker syndrome. We continued the medical workup and also treated the patient with psychotherapy. RESULTS: The patient's symptoms gradually abated, however, the headaches remained. No neurologic illness evolved during the follow-up. CONCLUSIONS: The diagnosis of pseudoseizures is suggested according to clinical evaluation and follow-up. A multidisciplinary approach is recommended in such cases for efficient diagnosis and treatment. PMID- 8711080 TI - Traumatic war experiences affect outcome in a case of Tourette's syndrome. AB - BACKGROUND: Stress has been associated with the course of Tourette's syndrome (TS). METHODS: The case report presented aims to emphasize the effect a severe traumatic experience has on the expression of TS. RESULTS: Following an intense traumatic experience, combat-related, a relatively mild case of TS became intractable. CONCLUSION: The combination of both of these severe, sometimes refractory, disorders is rare, and poses a therapeutic challenge. PMID- 8711082 TI - Behcet's disease and psychosomatic patterns of thinking. A single case study. AB - BACKGROUND: A single case of Behcet's disease was studied throughout several consultations during a period of 15 years. METHODS: The evolution of the disease was related to important events in the subject's life, cognitive patterns, psychopathological features and, particularly, his relationships to women and to his family. RESULTS: Some interesting complex correlations between illness course and psychosocial events and states were highlighted. CONCLUSIONS: Future prospective studies could better elucidate this multiple feedback. PMID- 8711083 TI - Serotonin, Cushing's disease, and depression. PMID- 8711084 TI - Serotonin receptors and their function in sleep, anxiety disorders and depression. AB - Thirty-three years ago, Gaddum and Picarelli classified the serotonin receptors in the guinea pig ileum into D and M types based on the activity of dibenzyline (D) and morphine (M) to block contractions of intestinal smooth muscle caused by serotonin. The subsequent location of specific ligand binding sites for serotonin in the brain has led to the identification of 14 serotonin receptor sub-types in rat brain. The cloning of these receptor sub-types has been of importance in enabling them to be classified as specific-protein molecules encoded by specific genes. The problem now arises with regard to the linking of the changes in the cellular activity of the various receptor sub-types with the plethora of behavioural changes that arise as a consequence of the actions of serotonin in the brain. The present review summarizes the evidence implicating the role of specific serotonin receptor sub-types in sleep, anxiety states, schizophrenia and depression. A summary of the relationship between these receptor sub-types and their possible involvement in the aetiology of schizophrenia, depression, anxiety and sleep disorders. PMID- 8711085 TI - Hypochondriasis influences quality-of-life outcomes in patients with chronic fatigue. AB - BACKGROUND: To determine how hypochondriacal symptoms influence the quality-of life outcomes of patients with a chief complaint of chronic fatigue. METHODS: Cross-sectional cohort study of a consecutive sample of 71 patients (mean duration of fatigue of 4.1 years). Forty-eight (68%) patients met criteria for current major depression and 32 (45%) met criteria for chronic fatigue syndrome (CFS). All patients received a comprehensive medical and psychiatric evaluation. Quality-of-life and physical, depressive and hypochondriacal symptom scores were assessed through reliable self-report questionnaires and a structured interview. A path model expressing the relation between predictor variables (hypochondriasis and depression), intervening variables (physical symptoms) and quality of life was postulated and evaluated using structural equation methods. RESULTS: The paths linking hypochondriasis with physical symptoms and mental health and the path connecting physical symptoms and quality of life were each statistically significant. The model applied especially well to patients who fulfilled CFS criteria. CONCLUSIONS: The quality of life of chronic fatigue patients correlates with the severity of their physical symptoms and their hypochondriacal disposition toward illness. PMID- 8711086 TI - Criterion-related validity of diagnostic criteria for alexithymia in a general hospital psychiatric setting. AB - BACKGROUND: The diagnostic criteria for alexithymia (DCA) was proposed by Fava et al. in 1995. The purpose of this study was to examine the criterion-related validity of the DCA in a general hospital psychiatric setting. METHODS: The subjects included 126 outpatients with psychiatric disturbance. The severity of alexithymic characteristics was assessed using the DCA and Beth Israel Hospital Psychosomatic Questionnaire (BIQ). Two Japanese versions of the 26-item Toronto Alexithymia Scale (TAS-26) and Minnesota Multiphasic. Personality Inventory-2 (MMPI-2) were administered. RESULTS: The DCA had relatively high sensitivity and specificity, indicating adequate discriminant validity. The discriminant validity of the DCA was also supported by significant associations of the DCA with the TAS 26 and MMPI-2. These results indicate that when used for screening, the DCA has adequate criterion-related validity. CONCLUSIONS: The DCA may be clinically useful in a general hospital psychiatric setting. PMID- 8711087 TI - Risk factors for suicide in multiple sclerosis. AB - BACKGROUND: The purpose of the present study was to identify risk factors for suicide in patients with multiple sclerosis (MS). METHODS: The study is based on available information about MS patients identified in the Danish MS Registry (DMSR) with onset in the period 1950-1985. We compared the MS suicides with the 1950-1985 onset cohort patients in the DSMR as to distribution of age at onset, presenting symptoms, and time from onset to diagnosis. We reviewed sociodemographic data, age of onset, the course of the disease, recent deterioration, type of deterioration, Kurtzke Disability Status Scale (DSS) score, previous mental disorder, type of mental disorder, previous suicide attempts, expression of suicidal intentions, circumstances at suicide, and suicide method for all MS patients who had committed suicide. In order to characterize MS suicides with respect to risk factors, comparisons were made for male and female suicides and for various groups of MS suicides according to disability status. RESULTS: The male suicide patients were characterized by a tendency to commit suicide in the age interval 40-49 years, by the use of a violent suicide method, by previous suicidal behaviour, by a previous mental disorder, by recent deterioration of MS, and by a moderate disability. For women the characteristics were less distinct. Patients with a severe course of MS had been subjected to more risk factors before the suicide than patients with a moderate course of the disease. CONCLUSION: Careful counselling and good information on all aspects of the disease, especially in the first stages and at time of progression, could be an instrument of prevention of suicides in MS patients. Furthermore, recognition and treatment of depression and pain is important. PMID- 8711088 TI - Mood changes in homesick persons during a holiday trip. A multiple case study. AB - BACKGROUND: Homesickness is generally studied with respect to rather uncommon long-lasting situations such as a (forced) move, while short stays away from home, like holidays, are much more common homesickness situations. Furthermore, little is known about the development and course of homesickness feelings. Therefore, the present multiple case study was designed in order to obtain a better insight into the onset and course of homesickness and patterns of mood changes, as well as saliva cortisol levels during a short stay away from home. METHODS: Several times before, during and after their holiday, 10 adult homesick prone females completed the Profile of Mood States (POMS), the Homesickness Decision Tree (HDT) and a Subjective Homesickness Rating. In addition, saliva samples were obtained. Data were analyzed individually. Furthermore, days with and without homesickness were compared across individuals using t tests. RESULTS: Using the HDT, the following typology, based upon four different reaction patterns, emerged: (1) no homesickness, (2) anticipation homesickness, (3) holiday homesickness, and (4) a combination of anticipation and holiday homesickness. POMS scores demonstrated significantly worsened mood when experiencing homesickness. Cortisol levels, however, failed to differ between days with and without homesickness. CONCLUSIONS: Homesickness is characterized by negative mood which is not necessarily reflected in significantly changed salivary cortisol levels. Feelings of homesickness are not only experienced during absence from home, but may occur also in anticipation to a leave. Future studies need to focus upon the prevalence, causes and consequences of the different types of homesickness. PMID- 8711090 TI - FDA shortened drug approval time in 1995. PMID- 8711089 TI - A cognitive-behavioral self-help educational program for patients with COPD. A pilot study. AB - BACKGROUND: The purpose of this pilot study was to evaluate the impact of an 8 week cognitive-behavioral self-help educational program for individuals with COPD on level of disability. METHODS: Eight adults ranging in age from 58 to 83 years attended a program that included a cognitive-behavioral intervention strategy. Levels of disability, cognitive distortion, anxiety, depression, somatic symptoms, and hostility were measured pre- and posttreatment using the Sickness Impact Profile, two Cognitive Error Questionnaires and the Symptom Questionnaire. RESULTS: Significant decreases were found in the Psychosocial and Total Sickness Impact Profile Scores (p value = 0.02 and p value = 0.03, respectively). On the General-Cognitive Error Questionnaire, the personalization subscale decreased significantly (p value = 0.04). On the COPD-Cognitive Error Questionnaire, the selective abstraction subscale decreased significantly (p value = 0.02). CONCLUSION: Findings suggest a positive treatment response to the program. PMID- 8711091 TI - New health care system can be built on expanded primary care. PMID- 8711092 TI - TB deaths reach historic levels. PMID- 8711093 TI - Anxiety disorders lead mental ills in United States. PMID- 8711094 TI - WHO warns of inadequate noncommunicable disease prevention. PMID- 8711096 TI - Epidemiologic maps of Washington DC, 1878-1909. AB - IN ORDER TO demonstrate the dire effects that contagious epidemic diseases were having on the population of Washington DC, Public Health Officers of the District of Columbia used epidemiological maps in their annual reports to city commissioners from 1878 until 1909. These maps charted information graphically that was included as statistical tables elsewhere in the reports. Their evolution occurred during a period when the Health Officers themselves were coming to grips with the causes of diseases that included diphtheria, scarlet fever, typhoid and malarial fevers, smallpox, consumption (tuberculosis), and diarrheal diseases. The maps may have encouraged laws that were passed during this period which required Washington DC physicians to report cases and deaths from these diseases. PMID- 8711095 TI - Hospitals and plastics. Dioxin prevention and medical waste incinerators. AB - CHLORINATED DIOXINS and related compounds are extremely potent toxic substances, producing effects in humans and animals at extremely low doses. Because these compounds are persistent in the environment and accumulate in the food chain, they are now distributed globally, and every member of the human population is exposed to them, primarily through the food supply and mothers' milk. An emerging body of information suggests that dioxin contamination has reached a level that may pose a large-scale, long-term public health risk. Of particular concern are dioxin's effects on reproduction, development, immune system function, and carcinogenesis. Medical waste incineration is a major source of dioxins. Polyvinyl chloride (PVC) plastic, as the dominant source of organically bound chlorine in the medical waste stream, is the primary cause of "iatrogenic" dioxin produced by the incineration of medical wastes. Health professionals have a responsibility to work to reduce dioxin exposure from medical sources. Health care institutions should implement policies to reduce the use of PVC plastics, thus achieving major reductions in medically related dioxin formation. PMID- 8711097 TI - Local research: needed guidance for the Indian Health Service's urban mission. PMID- 8711098 TI - Trauma among American Indians in an urban county. AB - OBJECTIVE: To describe severe injury among American Indians in a large metropolitan county given that most previous studies of the high Indian injury morbidity and mortality rates have been conducted primarily in rural areas. METHODS: A retrospective analysis of a hospital trauma registry was conducted for the years 1986-92 at the Harborview Medical Center, the only Level I trauma center in King County, Washington, metropolitan county with the seventh largest number of urban American Indians in the United States. RESULTS: Of 14,851 King County residents included in the registry, 593 (4%) were classified as American Indian. With King County whites as the reference, the age-standardized incidence ratio for inclusion of American Indians in the registry was 4.4 (95% confidence interval 4.1, 4.8). The standardized incidence ratios and proportional incidence ratios showed significant differences in mechanism and whether it was intentional or unintentional among Indians compared with whites. Hospitalizations for stab wounds, bites, and other blunt trauma were all significantly more frequent among Indians. Trauma admissions among Indians were disproportionately associated with assaults. A high proportion (72.3%) of American Indians tested had blood alcohol levels exceeding 0.1%. CONCLUSION: Urban American Indians experience high rates of trauma, differing from those among whites. Efforts to reduce injury in urban areas should include collaboration with representative urban American Indian organizations. PMID- 8711099 TI - Commentary on TB testing of inmates. PMID- 8711100 TI - Radiographic screening for tuberculosis in a large urban county jail. AB - OBJECTIVE: This study was designed to evaluate an innovative program of high speed radiographic screening for pulmonary tuberculosis (TB) at a large urban correctional institution, Cook County Jail in Chicago. METHODS: From March 1992 to February 1994, 126,608 inmates were screened on intake with a 100-mm mini chest radiograph. RESULTS: Sixty-seven cases of active TB were identified by radiograph and 19 others from diagnostic work-up. The case finding rate for active disease with radiographic screening was approximately double the rate previously achieved with Mantoux skin testing. Mean time from jail entry to isolation was reduced from 17.6 days with Mantoux skin testing to 2.3 days with radiographic screening. CONCLUSIONS: In large jail facilities, high speed X-ray screening for TB can minimize disruption of the intake process and lead to dramatic improvements in the efficiency of medical follow-up and isolation. PMID- 8711101 TI - Preventing perinatal transmission of HIV--costs and effectiveness of a recommended intervention. AB - OBJECTIVE: To calculate the national costs of reducing perinatal transmission of human immunodeficiency virus through counseling and voluntary testing of pregnant women and zidovudine treatment of infected women and their infants, as recommended by the Public Health Service, and to compare these costs with the savings from reducing the number of pediatric infections. METHOD: The authors analyzed the estimated costs of the intervention and the estimated cost savings from reducing the number of pediatric infections. The outcome measures are the number of infections prevented by the intervention and the net cost (cost of intervention minus the savings from a reduced number of pediatric HIV infections). The base model assumed that intervention participation and outcomes would resemble those found in the AIDS Clinical Trials Group Protocol 076. Assumptions were varied regarding maternal seroprevalence, participation by HIV infected women, the proportion of infected women who accepted and completed the treatment, and the efficacy of zidovudine to illustrate the effect of these assumptions on infections prevented and net cost. RESULTS: Without the intervention, a perinatal HIV transmission rate of 25% would result in 1750 HIV infected infants born annually in the United States, with lifetime medical-care costs estimated at $282 million. The cost of the intervention (counseling, testing, and zidovudine treatment) was estimated to be $ 67.6 million. In the base model, the intervention would prevent 656 pediatric HIV infections with a medical care cost saving of $105.6 million. The net cost saving of the intervention was $38.1 million. CONCLUSION: Voluntary HIV screening of pregnant women and ziovudine treatment for infected women and their infants resulted in cost savings under most of the assumptions used in this analysis. These results strongly support implementation of the Public Health Service recommendations for this intervention. PMID- 8711102 TI - Lead testing of children and homes: results of a national telephone survey. AB - OBJECTIVES: This study was designed to estimate the percentage of young children in the United States who have been tested for lead and the percentage of dwellings in the United States in which the paint has been tested for lead. METHODS: A national random digit dial telephone survey of 5238 households was conducted in 1994. Weighted national estimates and 95% confidence intervals for outcomes of interest were calculated. RESULTS: About 24% of U.S. children ages 0 to 6 years were estimated to have been tested for lead. Higher rates of testing were reported for children living in homes constructed prior to 1960, those living in homes with low household income, those living in rental units, and those living in the Northeast. Lead paint testing was performed for only an estimated 9% of U.S. housing units. Older homes were not more likely to have been tested than newer ones. CONCLUSION: A high proportion of pre-school children have apparently not been screened for lead exposure, even among subgroups at increased risk. Most dwellings of pre-school children have not been tested for lead paint. These data suggest that most at-risk children are not being reached by current approaches to lead poisoning prevention. PMID- 8711103 TI - Lesbian and bisexual women in small cities--at risk for HIV? HIV Prevention Community Collaborative. AB - Objectives.Women who have sex with women are a relatively hidden group that has been overlooked in most AIDS research and prevention efforts, primarily because the efficiency of HIV transmission between female partners is believed to be low. Although data are scant, it is commonly assumed that members of this population are not at high risk for HIV infection. However, a recent study of lesbian and bisexual women living in the San Francisco Bay area reported a relatively high seroprevalence rate and has raised additional questions about this group's HIV risk. The present study, the first to focus on lesbian and bisexual women living outside major AIDS epicenters, provides additional evidence. It describes risk factors for HIV transmission among lesbian and bisexual women living in small cities in four geographic regions of the United States.Methods.On three consecutive evenings in 1992, members of the research team distributed anonymous structured written surveys to women patrons as they entered gay bars in each of 16 small cities.Results.Almost 17% of bisexual respondents and 0.5% of lesbians reported having had unprotected vaginal or anal sex with a male during the two months prior to the survey. Almost 10 percent of bisexual women and 8.8% of lesbians in the sample reported a history of injection drug use. Among those women surveyed who said they had been tested, 1.4% reported they were infected with HIV.Conclusions.Self-identified sexual orientation was highly consistent with recent sexual behavior. HIV risk related to sexual behavior was concentrated among self-identified bisexual women. The prevalence of injection drug use was substantial among both bisexual and lesbians. Developers of HIV risk behavior programs should take the prevalence of these risk behaviors into consideration in the design of effective HIV prevention interventions tailored to the needs of this hidden population. PMID- 8711104 TI - Independent evaluation of the California Tobacco Education Program. AB - OBJECTIVE: To monitor the implementation of tobacco control programs and research in accordance with California's Proposition 99, approved by the voters in 1988, which increased the state's cigarette tax by 25 cents and designated one quarter of the increased revenue-approximately $100 million per year-to develop statewide media campaigns and to fund local health departments, community-based organizations, schools, and agencies working with high risk populations. METHODS: The authors evaluated the extent and the effectiveness of the implementation independently with standardized forms developed to track any tobacco-related information and activities, local media campaigns, changes in local policies and ordinances, training sessions, and prevention and cessation programs. Activities were reported on the forms to the authors monthly or quarterly. RESULTS: The evaluation indicated that local health departments, community-based organizations, and other group produced an exceedingly high volume of diverse tobacco control activities throughout the state. They reached a variety of ethnic groups and high risk populations with their programs. The data also show that local health departments and competitive grantees responded with an overall shift in their approach to tobacco control and moved from interventions focused primarily on individual people to broader, more community- and environment oriented interventions. Additionally, analysis of the wholesale tobacco tax revenues revealed that per capita purchasing continued to decline in California at a rate greater than in the rest of the United States. CONCLUSION: The evaluation demonstrated that Proposition 99 accomplished much of what it set out to do-reducing tobacco prevalence, reaching out to underserved populations, and heightening the awareness of the dangers of environmental tobacco smoke. PMID- 8711105 TI - Mortality among infants with congenital malformations, New York State, 1983 to 1988. AB - OBJECTIVE: The authors examined first-year mortality and risk factors for mortality among infants with major congenital malformations. METHODS: Infants with major congenital malformations born from 1983 to 1988 were identified from a statewide population-based congenital malformations registry. Variables analyzed included year of birth, birth weight, gestational age, infant sex, number of malformations, number of organ systems involved, level of care of the birth hospital, maternal age, maternal education, and maternal ethnicity. RESULTS: Infants with major malformations had a risk of death 6.3 times higher than the general population of live births. The risk declined from 6.5 in 1983 to 5.9 in 1988. Birth weight and number of malformations were the strongest risk factors. The likelihood of survival was similar for white and black infants. CONCLUSIONS: Being born with a malformation outweighs most of the other risks for infant mortality. Children with congenital malformations had higher cause-specific mortality for all causes except injury. PMID- 8711106 TI - Seasonal variation in weight-for-age in a pediatric emergency room. AB - OBJECTIVE: The authors assess seasonal variations in the prevalence of low weight for-age among young children visiting the pediatric emergency room of a city hospital. METHODS: We analyzed data on 11,118 children ages 6 to 24 months who visited the Boston City Hospital Pediatric Emergency Department between July 1989 and June 1992. Medical diagnoses were documented on a randomly selected subsample of 1,569 children. In addition, a questionnaire about food insecurity was administered to a convenience subsample of 269 families with children under 3 years of age. RESULTS: The percentage of children visiting the emergency room with weight-for-age below the fifth percentile was significantly higher for the three months following the coldest months than for the remaining months of the year, controlling for year of measurement. In the subsample, gastrointestinal illness was correlated with both season of measurement and weight-for-age, but the seasonal effect remained for the entire sample after controlling for dehydration. The questionnaire data suggested a relationship between economic stress and food insecurity that might help explain the seasonal effect. Families who were without heat or who were threatened with utility turnoff in the previous winter were twice as likely as other families to report that their children were hungry or at risk for hunger. CONCLUSIONS: Winter and early spring constituted periods of increased nutritional risk in the in this sample of predominantly low income children, probably because of the increased caloric associated with cold stress and infections. Further research is needed to assess whether decreased caloric availability due to high heating costs, a "heat or eat" effect, also contributes to this phenomenon. PMID- 8711107 TI - Perceptions of risks of drinking and boating among Massachusetts boaters. AB - OBJECTIVES: Public policy has treated drinking and boating as though it were analogous to drinking and driving. Accordingly, recent Federal and state laws to prevent drinking and boating have focused solely on alcohol use by the boat operator. This study was designed to determine boaters' knowledge about the epidemiology of boating fatalities and how boaters perceive the risks of drinking and boating. METHODS: In the summer of 1995, the authors conducted a survey by mail of a random sample of 600 owners of boats registered in Massachusetts. RESULTS: Survey results indicated that boaters believe passengers can safely drink more than operators. Respondents also thought that people on boats at rest can safely drink more than people on boats underway. CONCLUSIONS: The results of this study could be helpful in designing future boating safety campaigns by identifying gaps in knowledge about about the risks of drinking and boating for both operators and passengers. PMID- 8711108 TI - Creation of the National Institute of Mental Health. PMID- 8711109 TI - R.N. delivering baby with Dr.'s OK terminated. PMID- 8711110 TI - Failure to chart that Dr. was notified of pt.'s deterioration. Case in point: Sweeney v. Purvis 665 So. 2d 926--AL (1995). PMID- 8711111 TI - Legal case brief for nurses. AL: prosecution for practicing midwifery: license required for lay & nurse midwifery; GA: failure to chart time catheter removed: delay in reinsertion of catheter results. PMID- 8711112 TI - Wrong meds & death on "life flight": jurisdictional issues. Case in point: Cowart v. Shelby County. Health Care Corp. 911F. Supp. 248--MS (1996). PMID- 8711113 TI - Failure to heed nurses' request for exam: death results. PMID- 8711114 TI - Can a nurse be in two places at one time?: understaffing? Case in point: Merritt v. Karcioglu 668 So. 2d 469--LA (1996). PMID- 8711115 TI - Legal case briefs for nurse. TN: special shift request "unreasonable": workers' compensation benefits denied; SD: aides fail to report condition to nurses: respiratory arrest & brain damage result. PMID- 8711116 TI - Nurse has probable cause to detain & medicate pt. Case in point: Heater v. Southwood Psychiatric Center 49 Cal. Rptr. 880--CA (1996). PMID- 8711117 TI - Nurse reports sexual assault by Dr.: retaliatory termination. PMID- 8711118 TI - Nurse accused of failure to respond: "habit" testimony allowed. PMID- 8711119 TI - ND: "mooning" & sexual abuse of pt: "delay" in procedure not prejudicial. PMID- 8711120 TI - NY: elevator door slams on nurse attendant: +l.6 million judgment nullified. PMID- 8711121 TI - Don't hesitate to call for help: "when in doubt shout". PMID- 8711122 TI - Nurse ordered to "wean" patient from respirator: death. PMID- 8711123 TI - Lap pad left in patient: is nurse or doctor responsible? Case in point: Ravi v. Coates 662 So. 2d 218-AL (1995). PMID- 8711124 TI - Legal case briefs for nurses. AL: nurse has duty to "ensure" quality care: nurse does not "insure" patient's safety; OH: student dismissed under "catchall": courts to defer to faculty unless "arbitrary". PMID- 8711125 TI - Fetus born alive during abortion: Dr. orders extubation. Case in point: Hartsell v. Fort Sanders Reg. Med. Ctr. 905 S.W. 2d 944--TN (1995). PMID- 8711126 TI - Employees performing "ministerial" tasks have no immunity. PMID- 8711127 TI - Auto accident in traveling; "going-and-coming" rule. Case in point: Toolin v. Aquidneck Island Med. Resource 668 A. 2d 639--RI (1995). PMID- 8711128 TI - Legal case briefs for nurses. NY: violation of rehabilitation act alleged: nurse admits alcohol-related "blackouts"; CA: technician commits sexual assault on pt.: hosp. not liable under Respondent Superior. PMID- 8711129 TI - Posey restrained pt. chokes--brain damage: immunity issue. Case in point: Allen v. Montgomery Hosp. 668 A. 2d 565--PA (1995). PMID- 8711130 TI - The pedunculopontine nucleus--auditory input, arousal and pathophysiology. AB - This review describes the role of the pedunculopontine nucleus (PPN) in various functions, including sleep-wake mechanisms, arousal, locomotion and in several pathological conditions. Special emphasis is placed on the auditory input to the PPN and the possible role of this nucleus in the manifestation of the P1 middle latency auditory evoked response. The importance of these considerations is evident because the PPN is part of the cholinergic arm of the reticular activating system. As such, the auditory input to this region may modulate the level of arousal of the CNS and, consequently, abnormalities in the processing of this input can be expected to have serious consequences on the level of excitability of the CNS. The involvement of the PPN in such disorders as schizophrenia, anxiety disorder and narcolepsy is discussed. PMID- 8711131 TI - Apoptosis in the developing CNS. AB - In this review, apoptosis during normal development of the CNS and abnormal apoptosis inducing hydrocephaly and arhinencephaly will be discussed. As the prominent sites of apoptosis during normal development of the CNS, we focused on the area of fusion of the neural plate to form the neural tube, the developing rhombomeres, and neuronal loss in the CNS during embryogenesis and postnatal development. As examples of abnormal apoptosis inducing abnormal brain morphogenesis, we will discuss genetically induced arhinencephaly and hydrocephaly. It was suggested that apoptosis of the precursor mitral cells in the anlage of the olfactory bulb was induced by non-innervation of olfactory neurons, and apoptosis of the precursor neurons in the pyriform cortex was induced by the non-innervation caused by the death of mitral cells in the mutant arhinencephalic mouse brain (Pdn/Pdn). Thus, sequential apoptosis of the precursor neurons and sequential manifestation of the brain abnormalities were proposed in arhinencephalic mutant mouse embryos and also in the arhinencephalic brains induced experimentally by fetal laser surgery exo utero. Meanwhile, it was speculated that the Gli3 gene, mutation of which is responsible for the arhinencephaly in Pdn/Pdn mice, might play a role in mesenchymal programmed cell death during development. PMID- 8711132 TI - The temporal relationship between the brainstem and primary cortical auditory evoked potentials. AB - Many methods are employed in order to define more precisely the generators of an evoked potential (EP) waveform. One technique is to compare the timing of an EP whose origin is well established with that of one whose origin is less certain. In the present article, the latency of the primary cortical auditory evoked potential (PCAEP) was compared to each of the seven subcomponents which compose the brainstem auditory evoked potential (BAEP). The data for this comparison was derived from a retrospective analysis of previous recordings of the PCAEP and BAEP. Central auditory conduction time (CACT) was calculated by subtracting the latency of the cochlear nucleus BAEP component (wave III) from that of the PCAEP. It was found that CACT in humans is 12 msec which is more than double that of central somatosensory conduction time. The interpeak latencies between BAEP waves V, VI, and VII and the PCAEP were also calculated. It was deduced that all three waves must have an origin rather more caudally within the central auditory system than is commonly supposed. In addition, it is demonstrated that the early components of the middle latency AEP (No and Na) largely reside within the time domain between the termination of the BAEP components and the PCAEP which would be consistent with their being far field reflections of midbrain and subcortical auditory activity. It is concluded that as the afferent volley ascends the central auditory pathways, it generates not a sequence of high frequency BAEP responses but rather a succession of slower post-synaptic waves. The only means of reconciling the timing of the BAEP waves with that of the PCAEP is to assume that the generation of all the BAEP components must be largely restricted to a quite confined region within the auditory nerve and the lower half of the pons. PMID- 8711133 TI - Induction of both cytosolic phospholipase A2 and prostaglandin H synthase-2 by interleukin-1 beta in WISH cells in inhibited by dexamethasone. AB - In previous studies we have shown that IL-1 beta induced both PGE2 release and total cellular cPLA2 activity and cPLA2 protein synthesis in human amnion-derived WISH cells. In this study, the effect of IL-1 beta on cPLA2 and PGHS-2 mRNA expression was investigated. Using RT-PCR, we found that IL-1 beta (0.1 ng/ml) coordinately induced both cPLA2 and PGHS-2 mRNA expression within 2 hours. The synthetic glucocorticoid dexamethasone (10(-10)-10(-6)M) inhibited IL-1 beta induced cPLA2 and PGHS-2 mRNA expression activity and protein synthesis and PGE2 release in a concentration dependent manner. In the absence of IL-1 beta, dexamethasone alone (10(-6)M) inhibited basal cPLA2 activity, mRNA expression and protein synthesis. In addition, cycloheximide (5 micrograms/ml) apparently superinduced, but actinomycin D (2 micrograms/ml) inhibited IL-1 beta-induced cPLA2 and PGHS-2 mRNA expression suggesting that both are immediate early genes and a transcriptional mechanism is involved in the induction of both cPLA2 and PGHS-2 mRNA by IL-1 beta. PMID- 8711134 TI - Alterations of intrauterine eicosanoid production in pregnancy-induced hypertension: decreased production of 12-hydroxyeicosatetraenoic acid in the placenta. AB - The important role of eicosanoids in pregnancy-induced hypertension is generally accepted. Because of the lack of innervation of the uteroplacental vessels, humoral vasoactive factors are important for the regulation of vascular tone. Until now, mainly the balance of vasodilatative and vasoconstrictive prostaglandins has been studied. We were able to confirm their intrauterine imbalance in hypertensive pregnancies. In addition, the placental production of less known lipoxygenase metabolites has been analyzed in this study. Intrauterine tissues (30-100mg wet weight) were examined for their release of eicosanoids. Short term tissue cultures were performed in Hanks balanced salts solution (HBSS) at 37 degrees C in an atmosphere of 95% air/5% CO2 with and without incorporation of tritiated arachidonic acid. The arachidonate metabolites in culture media were analyzed by High Performance Liquid Chromatography (HPLC) with radioactivity detection or by enzyme immunoassays or radioimmunoassays, respectively. All intrauterine tissues released more lipoxygenase metabolites than cyclooxygenase metabolites with 12-hydroxy-eicosatetraenic acid (12-HETE) as their main metabolite. The placental release of 12-HETE was significantly decreased in hypertensive pregnancies. In hypertensive pregnancies the ratio TXB2/6-keto-PGF1 alpha synthesis was increased. Lipoxygenase metabolites, especially 12-HETE, seem to have important physiological and pathophysiological functions in the intrauterine compartment. Their biological role in this context needs further investigation. PMID- 8711135 TI - Effect of a novel leukotoriene synthesis inhibitor, BAY x1005, on the antigen-and LPS-induced airway hyperresponsiveness in guinea pigs. AB - Due to the inhibition of 5-lipoxygenase-activating protein (FLAP), BAY x1005 is a new selective inhibitor of leukotriene synthesis. The effects of BAY x1005 on the antigen- and bacterial lipopolysaccharide (LPS)-induced airway hyperresponsiveness in guinea pigs were investigated. Six times provocation of aeroantigen caused biphasic increases in airway resistance which peaked at 1 hr (immediate phase reaction) and 4 hrs (late phase reaction). It also caused airway hyperreactivity to acetylcholine. BAY x1005 at doses of 10 mg/kg and 30 mg/kg significantly inhibited antigen-induced increase in respiratory resistance (Rrs) at 1 and 4 hrs after the last antigen challenge. Simultaneously, BAY x1005 inhibited the antigen-induced airway hyperresponsiveness at doses of 10 and 30 mg/kg and airway eosinophilia (bronchoalveolar lavage study) at a dose of 30 mg/kg. In addition, BAY x1005 at a dose of 30 mg/kg inhibited bacterial LPS induced airway hyperreactivity to acetylcholine. In this model, BAY x1005 did not affect the increase of the number of leukocytes in bronchoalveolar lavage fluid. These results suggest that BAY x1005 is a potent anti-asthmatic agent with an inhibitory action to airway hyperreactivity. PMID- 8711136 TI - Antithrombotic effect of topically applied 3-oxa-methano-prostaglandin I1 in the microcirculation and antiplatelet functions of its active form. AB - The antithrombotic effect of topical application of the 3-oxa-methano prostaglandin (PG) I1 analog, SM-10902 in the microcirculation and in vitro antiplatelet functions of its active form SM-10906 were estimated in comparison with PGI2 and PGE1. In rat platelets, SM-10906 evoked accumulation of intracellular cyclic adenosine 3',5'-monophosphate, and exhibited antiaggregatory and disaggregatory activities, which were all enhanced by the phosphodiesterase inhibitor theophylline. Additionally, SM-10906 was shown to inhibit platelet adhesion to collagen in human platelet-rich plasma. PGI2 and PGE1 also showed in vitro antiplatelet effects in the order of PGI2 > SM-10906 > or = PGE1. SM-10902 exhibited a dose-dependent anti-thrombotic effect in the guinea pig mesenteric arteriole by a topical application, and this activity might be exerted by the antiplatelet functions of SM-10906. Although SM-10906, PGI2 and PGE1 also showed the antithrombotic effects, SM-10902 was the most potent. In conclusion, the present studies indicate that an external topical preparation of SM-10902 may be useful for the therapy of peripheral circulatory insufficiency. PMID- 8711137 TI - Relationship between prostaglandin in peritoneal fluid and pelvic venous congestion after sterilization. AB - Pelvic congestion may cause chronic pelvic pain in women. The aim of the study is to elucidate a possible role in this condition for prostaglandin. Prostaglandin levels in peritoneal fluid were measured in 18 women with pelvic pain caused by pelvic congestion following sterilization, 10 women without pain following sterilization, and 10 normal healthy women. Peritoneal fluid was aspirated by a silastic catheter from the cul-de-sac under laparoscopic direct vision. Concentration of 6-keto-PGF1 alpha, TXB2, PGF2 alpha and PGE2 were measured with the standard radioimmunoassay method in all samples. Results showed that 6-keto PGF1 alpha levels in peritoneal fluid from patients with pelvic congestion were markedly higher than those from two control women (P < 0.05); 6-keto-PGF1 alpha/TXB2 in pelvic congestion and control groups were markedly different (P < 0.05); the total amounts of peritoneal fluid was higher in pelvic congestion than that in two control groups (P < 0.001). These data suggested that 6-keto-PGF1 alpha is increased in peritoneal fluid of women with pelvic congestion and the change might play an important role in attack of this disease. PMID- 8711138 TI - Genistein suppresses EGF-induced prostaglandin biosynthesis by a mechanism independent of EGF receptor tyrosine kinase inhibition. AB - This study demonstrated that genistein, a selective tyrosine kinase inhibitor, blocked PGE2 production in human A431 and WISH cells and murine 3T3 cells in response to epidermal growth factor and platelet-derived growth factor. Blockade of growth factor-induced PGE2 production was dose-dependent (IC50 approximately equal to 7-8 microM). Genistein also abolished PGE2 formation in response to calcium ionophores, A23187 and ionomycin, and the phorbol ester, phorbol myristate acetate. Moreover, genistein-treated A431 and WISH cells incorporated significantly less [3H]arachidonic acid into membrane phospholipids than control cells. Finally, genistein decreased the specific activity of prostaglandin H2 synthase prepared from A431 cells, WISH cells, and ram seminal vesicle. The IC50 of genistein for inhibition of prostaglandin H2 synthase specific activity extracted from A431 and WISH cells approximated that half-maximal inhibitory concentration in the whole cell assay. These data indicate that genistein may interfere with arachidonic acid metabolism at several key points by a mechanism(s) that is independent of its inhibitory action on receptor tyrosine protein kinases. Taken together, these results also suggest that caution should be exercised when drawing conclusions about the putative role of tyrosine kinases in signal transduction events using genistein as a pharmacological blocker. PMID- 8711139 TI - [Rubella in 1994]. PMID- 8711140 TI - [Cerebrospinal meningitis and encephalitis in 1994]. PMID- 8711141 TI - [Salmonellosis in 1994]. PMID- 8711142 TI - [Bacterial dysentery in 1994]. PMID- 8711143 TI - [Food poisoning and infection in 1994]. PMID- 8711144 TI - [Botulism in 1994]. PMID- 8711145 TI - [Humoral response in hepatitis C infections: types of anti-HCV and reactivity evaluated using BLOT-index]. AB - In this paper an attempt has been made at quantitative evaluation of reactivity of various types anti-HCV using proposed BLOT-index. We based our research on 20 patients diagnosed with hepatitis-C (clinical, biochemical, serological and enzymatical criteria--repeatedly positive results of second generation screening tests EIA ABBOTT HCV and UBI HCV Monoelisa Organon Teknika) who were anti-HCV determined at the beginning the acute phase and after 3 and 6 months. Multiantigen tests were used veryfying LIATEK-HCV 2 and LIATEK-HCV 3 Organon Teknika. Value of BLOT-index was algebraic sum of "pluses" for particular anti HCV. Three models of dynamics of BLOT-index were observed: increase, plateau and decrease. Statistically significant differences between values of BLOT-index were shown at the beginning of the acute phase of hepatitis-C and after 6 months. There were no differences observed between convalescents and patients who developed chronic hepatitis-C (test t-Student's and f-Fisher's). Reactivity of anti-HCV, evaluated using BLOT-index, shown increasing trend, which is a dominant pattern, however quantitative and qualitative changes do not occur more frequently than every 6 months. The LIATEK-HCV 3 gives fewer undetermined reactions and detects anti-HCV earlier than a former generation of this test. PMID- 8711146 TI - [Human poisoning caused by chemicals for plant protection in 1994]. PMID- 8711147 TI - [Viral hepatitis B in 1994]. PMID- 8711148 TI - [Viral hepatitis non-B in 1994]. PMID- 8711149 TI - [Tetanus in 1994]. PMID- 8711150 TI - [Rabies in 1994]. PMID- 8711151 TI - [Brucellosis in 1994]. PMID- 8711152 TI - [Trichinosis in 1994]. PMID- 8711153 TI - [Taeniasis in 1994]. PMID- 8711154 TI - [AIDS and HIV infections in 1994]. PMID- 8711155 TI - [Herpes simplex encephalitis in children]. AB - Clinical course of Herpes Simplex Encephalitis were shown in this paper. 24 children in the age of 4 to 15 were treated at Polish Mother's Memorial Hospital in years 1992-1994. Value of EEG, CT scan, NMR were underlined in early diagnostic and casual treatment of diseases. All children from this analysis group survived, only 8 demonstrated late complications: epileptic attacks (3), spastic tetraparesis (1), encephalopathy and mental disturbances (1). Positive results of treatment were connected with early diagnosis and early antiviral treatment which is very important in decreasing death rate and preventing permanent injury. PMID- 8711156 TI - [Vaccination against hepatitis B]. AB - Current epidemiological problems, world strategy for vaccination as outlined in the Expanded Programme on Immunization of the WHO, structure of the hepatitis B virus, and vaccine-escape mutants were summarized briefly in the first part of the review. Later, generation of plasma-derived and recombinant vaccines containing S antigen only, and subsequently, S together with preS1 + preS2 antigens were described. The results of vaccination of adults and children with high risk of infection at two symposia held in our country in 1991 and 1994, were analysed. The role in prevention of the spread of viral hepatitis of high hygienic standard in all health care units and education of staff in terms of anti-infectious control was pointed out. Finally, the problem of persistence of anti-HBs antibodies in the years following the immunization was outlined. PMID- 8711157 TI - [Prophylaxis of influenza]. PMID- 8711158 TI - [The incidence of virus respiratory infections in small children in the season 1994-95]. AB - As previously, by antibody fluorescent technique the etiology of viral respiratory infections was determinated in small children hospitalized because of respiratory disease in the epidemic season 1994-95. Data obtained were compared with the results from the preceding season. Among a total of 371 patients, virus infection was detected in 170 (45,8%). Data for the whole season did not distinctly differ from those obtained previously; however, some differences were found in the occurrence of the individual virus infections in consecutive months of the epidemic season. The second (spring) wave of RS virus infections in this season was lower and more extended in time than in the previous seasons. Highest incidence of adenoviruses was observed in July 1995. The incidence of parainfluenza type 4 virus was higher in the disease of small children. In the youngest (below 1 month) age group percent of virus infections was higher, as compared with data from the preceding season. PMID- 8711159 TI - [The immunochemical characteristics of adult sera after Td vaccination]. AB - The results of immunochemical test of 29 adults sera before and after Td vaccination were presented. The level of 12 protein fractions was in normal range value in 26 of examined sera. There was IgA deficiency in one serum, in two others haptoglobin deficiency and high levels of transferrin and IgM were observed. PMID- 8711160 TI - [Polysaccharide vaccines. I. Pneumococcal vaccines]. PMID- 8711161 TI - [The presence of antibodies against Francisella tularensis among inhabitants of North-Eastern Poland]. AB - In this work we analysed the frequency of incidence of antibodies against F. tularensis among forest workers in region of North-Eastern Poland. The presence was detected in 0,13% analised workers and 3,38% hospitalised patients in Clinic of Parasitic Diseases and Neuroinfections in Bialystok. PMID- 8711162 TI - [Frequency of birth defects in four different regions in Poland]. AB - The aim of this paper is preliminary assessment of birth defect in different regions of Poland with various environmental pollution. During the first 3 month of registration among 7253 newborns were 91 (1,3%) birth defect (0,8%-1,6% in different regions). PMID- 8711164 TI - [Measles in 1994]. PMID- 8711163 TI - [Infectious diseases in Poland in 1994]. PMID- 8711165 TI - [Whooping cough in 1994]. PMID- 8711166 TI - [Scarlet fever in 1994]. PMID- 8711167 TI - [Mumps in 1994]. PMID- 8711168 TI - [Influenza in 1994]. PMID- 8711169 TI - [Bronchial asthma--a chronic inflammatory disorder]. AB - Inflammation is a major process in the pathogenesis of bronchial asthma. Pivotal role in the induction of the inflammation in atopic subjects is played by mast cells and eosinophils and their mediators. Lymphocytes T and macrophages modulate this process. Although the pathogenesis of asthma in non-atopic subjects is not totally clear the inflammation has similar course as in asthma with IgE overproduction. Current research emphasise e.g. the role of adhesion molecules, bronchial epithelial cells and nitric oxide in the pathogenesis of asthma. Selectin E, ICAM-1, VCAM-1 take part in the migration of inflammatory cells in the asthmatic lung. Expression of these molecules is included by IL-1, TNF-alpha, and IL-4. Inflammation leads to the bronchial epithelial damage and release of proinflammatory cytokines, which augment the above process. The epithelial damage causes exposure of nerve endings, which can lead to the activation of axon reflexes. The concentration of NO in the exhaled air of asthmatics is much higher than in healthy subjects. It may be produced by inflammatory cells and may augment the inflammation as well as cause bronchial hyperresponsiveness. The better understanding of inflammatory patterns of bronchial asthma has major influence on the therapeutic approach. Inhaled anti-inflammatory drugs are of the first choice in pharmacotherapy of even mild forms of asthma. PMID- 8711170 TI - [Monoclonal antibodies in diagnosis of acute leukemias]. AB - Immunophenotyping has become an essential component for the study of acute myeloblastic (AML) and lymphoblastic (ALL) leukaemias. The recent development of highly specific monoclonal antibodies (Mc Ab) to differentiation antigens (CD) of haematopoetic cells have made it readily available to clinical laboratories in most major hospitals. Immunophenotyping complements standard morphology by providing information on lineage, stage of differentiation and clonality. In addition some of the flow cytometry findings have independent prognostic significance. Monoclonal antibodies useful in defining lineage (B-cell versus T cell) and stages of differentiation of ALL. It can be also used in identifying characteristic feature of AML and aiding in lineage determination in acute leukaemias that are morphologically undifferentiated. Surface immunophenotyping is especially helpful for recognizing mixed lineage acute leukaemia and diagnosing certain rare entities such as erythroleukaemia (M6), acute megakaryocytic leukaemia (M7) and minimally differentiation acute myeloid leukaemia. PMID- 8711171 TI - [Physiologic principles of osteoporosis]. PMID- 8711172 TI - [Vitamin disturbances in chronic renal insufficiency. II. Fat soluble vitamins]. AB - Fat soluble vitamins (except vitamin K) are protein bounded with subsequent storage in the body. It is generally accepted that plasma level of vitamin A is increased in majority of patients with chronic renal insufficiency (CRI) including those on continuous ambulatory peritoneal dialysis (CAPD). Thus, there is no need to supplement this vitamin in CRI patients (pts). Plasma level of vitamin E in CRI pts may be elevated, normal or decreased. It seems to be justified to supplement this vitamin, in spite of its normal plasma level, in case platelet aggregation is increased. Both in dialyzed and nondialyzed CRI pts a normal serum level of vitamin K has been observed. Decreased or extremely low serum level of vitamin D following the gradual loss of renal tissue is to be observed in CRI pts. This deficit is regarded as the main factor leading to the decrease in serum level of calcium, the secondary hyperparathyroidism and bone changes. Treatment with 1.25(OH)2D3 (calcitriol) proved to be most successful in alleviation of symptoms resulting from the deficit of vitamin D3 in the body. Intravenous "pulsating" administration of calcitriol results in rapid normalization of serum PTH level. Treatment with 25(OH)D3 (calcidiol) given orally 3 times a week ("pulsating" method) revealed also fairly good results in this respect. During treatment with vitamin D3 hypercalcemia tends to develop, serum alkaline phosphatase normalizes, elevated PTH serum level decreases. Vitamin D metabolites are less active than 1.25(OH)2D3 being less hypercalcemic. PMID- 8711173 TI - [Diabetic foot syndrome in clinical practice. I. A model of a Diabetic Foot Consulting Unit]. AB - Complex pathogenesis of the foot syndrome involving a number of body system and tissues provided us with grounds for making multispecialistic assessment of its severity. Also a decision about the type and intensity of procedure should be multispecialistic. An interdisciplinary team consists of a diabetologist, diabetic educator, podiatrist, kinesitherapist, orthopedist, general and vascular surgeon and a consultant from manufacturing corrective shoes. The make-up of the team implies that the procedure includes education, metabolic normalization of diabetes mellitus, orthopedic correction, possible surgical intervention and rehabilitation. Results discussed in part II of the study indicate the effectiveness of such approach. PMID- 8711174 TI - [Contemporary opinions on the use of aspirin, heparin and thrombolytic therapy in unstable angina pectoris--pathogenetic findings versus results of many clinical tests]. AB - We summed up the results of clinical researches on thrombolysis in unstable angina pectoris (UAP). We also presented results of similar researches on aspirin and-or versus-heparin in UAP. The idea of applying thrombolytic therapy in UAP was based on data of anatomopathologic, coronary angiographic, biochemical and recently intracoronary ultrasonographic findings. These data indicated that the acute coronary syndromes (UAP, non-Q wave and Q-wave myocardial infarction) were very often caused by coronary plaque disruption and thrombosis. Although the efficiency of thrombolytic therapy in UAP was confirmed angiographically, nevertheless thrombolysis did not significantly decrease in-hospital cardiac events. Bleeding complications were significantly higher in patients who had received thrombolytic therapy. Heparin in constant i.v. infusion and aspirin administered orally seemed to improve clinical outcome. Thrombolytic therapy is not recommended in patients diagnosed as having unstable angina pectoris. PMID- 8711175 TI - [Vascular factors in the mechanism of acute pancreatitis]. AB - Clinically, acute pancreatitis can sometimes be seen in local or systemic vascular diseases. Vascular mechanisms of acute pancreatitis are defined as impairment of pancreatic blood inflow and outflow, or disturbance of the pancreatic microcirculation. Pancreatic macro-circulatory flow can be impaired by low flow states such as hypovolemic shock, cardiogenic shock, or by occlusion of pancreatic arteries or veins. The pancreatic microcirculation can be compromised by embolization of microvessels or by induction of endothelial lesions. It is known that pancreatic ischemia is an initiating or promoting factor of acute pancreatitis. It has been proposed that impaired local pancreatic microcirculation results from leukocytes-endothelial cell adhesion and presence of various vasoactive substances or chemical mediators such as histamine, serotonin, prostaglandins, thromboxanes, leukotrienes, platelet-activating factor, endothelin, free radicals, kinins and adhesion glycoproteins. PMID- 8711176 TI - [Diabetic foot syndrome in clinical practice. II. Results after one year at the Diabetic Foot Consulting Unit]. AB - The study included 46 patients with the diabetic foot syndrome and ulcerations, and 40 patients with high-risk foot. Mean duration of hospitalization of the patients with ulcerations was 54 days, mean daily glycemia decreased from 162.5 mg% to 114.9 mg%. Ulcerations were completely healed in 93.5% of patients, whereas high and partial foot amputation was high and 3.2% of patients, respectively. The annual amputation rate was 2.3% in the entire group of diabetic foot patients (86). The present results indicate the necessity for adoption of a multidisciplinary approach to the problem of diabetic foot. It may be added that cooperation in glycemia normalization and patients education may decrease the number of amputations, and hence the degree of physical disability in diabetic foot patients. PMID- 8711177 TI - [Comparison of calcitonin tolerance after intramuscular or intranasal administration in treatment for postmenopausal osteoporosis]. AB - The aim of the study was to compare the tolerance of synthetic salmon calcitonin applied in two different ways, intramuscularly or intranasally, in 50 patients with postmenopausal osteoporosis with bone fractures. Thirty patients were treated with calcitonin in intramuscular or subcutaneous injections, whereas in 20 cases calcitonin was applied in nasal spray. In the first group several side effects were observed, like nausea, vomiting, abdominal pains, skin rush, headaches, dropping blood pressure, symptoms of bronchial spasm. Finally in 13 cases it was necessary to stop calcitonin therapy. On the other hand the patients receiving calcitonin in nasal spray did not manifest any severe side effects. PMID- 8711178 TI - Current problems of clinical toxicology in Poland. PMID- 8711179 TI - Special features of chemical accidents. PMID- 8711180 TI - Emergency procedures in the event of chemical exposure/accidents. PMID- 8711181 TI - The "EMER.CHEM" project: computerised management of hazardous materials incident. PMID- 8711182 TI - [Selected chemical compounds as potential sources of massive poisoning. Physicochemical properties, poisoning symptoms, treatment]. AB - Characteristics of 42 chemical compounds (17 gases and 24 liquids) sustaining potential danger of massive poisonings in case of accident or chemical disaster is presented. Description of each substance includes: physiocochemical properties, pathophysiology and symptoms of acute poisonings with those chemicals. Rules and procedures of treatment are presented including premedical first aid and medical treatment. PMID- 8711183 TI - The usefulness of scintigraphic examination for the evaluation of hepatotoxic impact of cholinesterase inhibitors. AB - The aim of the study was to evaluate the morphological and functional status of the liver in acute, oral cholinesterase inhibitors poisoning using static scintigraphy, hepatography and measurements of chosen enzymes activity. Considering the different clinical picture of cholinesterase inhibitors poisonings in people, it was necessary to estimate the poisoning severity and its dependence on the frequency and intensification of the liver lesion. Under examination there were 37 cholinesterase inhibitors orally poisoned patients, treated at the Department of Toxicology in the years 1992-1995. The examined group comprised 7 women (19%) and 30 men (81%). Organophosphate compounds poisoning was noted in 14 patients, and carbamates poisonings in 23 patients. The reference group comprised 30 healthy men aged 24 to 59 years not exposed to hepatotoxic agents. More than 90% of patients were classified as severe poisoned. Any fatal case was not noted. A differently intensified pathological changes of the liver dependent on age and poisoning severity were found in 97.2% of patients and their frequency was significantly higher than in the control group. Hepatographic picture revealed in 96.6% of cases the liver lesion. Hepatographic picture of the liver was also dependent on poisoning severity. The higher activity of AST, ALT, AP and higher bilirubin concentration in blood were noted in poisoned men compared to the control group. Control scintigraphic examination revealed a considerable improvement in the intensification of the liver scintigraphic picture in 40% of the patients and a higher intensification in 13% of the subjects. In 46.6% of the patients the intensification of scintigraphic changes remained at the same level. Considering arbitrary criteria for the degree of the liver lesion, the improvement in the intensification of hepatographic changes was noted in 42.8% of the patients; the intensification of the liver lesion was not noted even in one case. Analyzing the percentage of the liver lesion for each individual patient, improvement was noted in 92.8% of the examined patients, and the changes with the same level of intensification in 7.2%. Deterioration was not noted at all. CONCLUSION: The liver scintigraphy and hepatography combined with biochemical analysis allows to assess the liver condition in acute cholinesterase inhibitors poisoning. PMID- 8711184 TI - Acute ethanol poisonings during the New Year's Eve party "95/'96 in Krakow. AB - Distilled spirits are the major source of ethanol consumption in Poland. Such concentrated alcohol is drunk quickly, especially in teenager groups who consider consumption of 250 or 500 ml of vodka during one hour or one hour and half as quite normal. For two years the New Years Eve party has been celebrated in the Main Market Square in Krakow. In 1995 unfavourable weather conditions had been observed for at least 14 days before the New Year's Eve night. During that night only 15 people-participants of the party were admitted to the Department of Toxicology. The aim of this study is to present the cases of poisoned people with ethanol during the last night of the year 1995. All the patients were young (10 poisoned patients were below 17 years of age), considerably low body temperature, and relatively low coma grade. 80% of the patients were admitted between 0:00 and 5:00 a.m. after 200 minutes since the moment they started to drink alcohol. Blood ethanol concentration ranged from 1.2 to 4.23 g/L and serum osmolality from 338 to 440 mOsm/kg H2O. Both ethanol concentration and serum osmolality progressively rose reaching the highest values in the group of patient with III coma degree (p < 0.001). All the cases of acute ethanol poisoning presented in the study had a good outcome, as patient were admitted at the Clinic in relatively short time after symptoms of poisoning have occurred. PMID- 8711185 TI - Causes of poisoning in children during the period 1990-1993. AB - We retrospectively analysed the files from our Centre concerning intoxication in children younger than 14 years. About 96% of intoxication were accidental, 43% of patients were poisoned with drugs, 22% with house-hold products and 15% with pesticides. Almost 25% of drug poisonings were due to sedative and to psychotropic drugs, the 20% to drug mixtures. The frequency of mushrooms and plants poisonings were low-7%. In Poland there are no acute poisoning departments which treat children, and thus these patients are treated in paediatric wards and paediatric hospitals, so complete data on the poisonings in children from over the whole territory of Poland are not available (due to the great number of paediatric wards in Poland and to the lack of a computer network, collection of such complete data is practically impossible). However, we believe that the frequency of causes of children's intoxication in our country is properly mirrored in our material. This allows conclusions to be drawn the prophylaxis of poisoning in this age group. PMID- 8711186 TI - Acute poisonings in the course of drug addiction: chemicotoxicological diagnostics. AB - A demand for a quick and reliable diagnostics of acute and chronic poisonings with drugs of abuse, monitoring of the treatment of drug abusers and controlling their abstinence lead to a search for the most useful diagnostic methods for clinical aims and certification. Our observations concerning the importance of toxicological investigation, particularly the use of Triage rapid immunoassay, in the diagnostics of acute poisonings and the abstinence control of dependent people were presented. Urine of 418 poisoned people and 7 "detoxificated' patients treated at the Department of Toxicology in Krakow in the years 1991-1995 was examined, and in 54 cases Triage method was used (seven drugs of abuse were determined: amphetamine, barbiturates, benzodiazepines, cocaine, methadone, opiates and tetrahydrocannabinoids), which was later confirmed using the Fluorescence Polarisation immunoassay, measured with the Vitalab Eclair. An increase in number of admitted patients to the Department of Toxicology was noted and the need for reliable diagnostics concerning the increasing number of cases in which inconsistency of the drug declared in the interview with the toxicological findings was stated. Triage method used in the examinations proved to be very useful for clinical diagnostics in quick determination of drugs of abuse in urine. The result should be confirmed using another, preferential method for the poison found in biological material. PMID- 8711187 TI - The usefulness of carboxyhemoglobin, methemoglobin and blood lactate concentration in evaluating the health condition of Krakow inhabitants exposed to primary pollutants. AB - Determination of the carboxyhemoglobin (COHb), methemoglobin (MetHb), and blood lactate concentrations is known as important contributive in evaluation of oxygen deficiency in people from urban populations. The methemoglobinemia is usually caused by unfavourable impact of NOx. A quantification of COHb allows, to some extend, to evaluate the threat of population with carbon monoxide (CO). An elevation in the blood lactate concentration indicate the oxygen insufficiency of the examined patients. The aim of this study was to evaluate the concentrations of the mentioned parameters in the three groups Krakow's population differently exposed to CO produced by incomplete combustion of carbon-containing materials. The group I consisted of 53 patients treated at the Toxicological Department for acute CO poisoning. The group II comprised 31 subjects professionally exposed to CO. The group III comprised 99 adult Krakow's inhabitants who started education in the technical school and had never been professionally exposed to CO (control group). The elevation in methemoglobin concentration was not found in any of the examined group. The highest COHb level was noted in the group of the acute CO poisoned patients. No significant difference between COHb levels was found in the chronically exposed group (II) and in the control one. The increased blood lactate concentrations were found in both the acutely CO poisoned and in the group of chronically, by occupation, exposed to CO. No unfavourable impact of Nox reflected in methemoglobinemia was noted in any examined population. The determination of COHb levels is helpful in diagnosis of acute CO poisoning. The measuring of the blood lactate concentration allows to detect the oxygen insufficiency in acute CO poisoning and in states of long-term exposure to CO. PMID- 8711188 TI - Prenatal lead exposure and the pregnancy outcome. A case-control study in southern Poland. AB - In an attempt to test null hypothesis (Ho): that prenatal lead exposure does not increase the risk of prematurity and the delivery of SGA infants, a case-control study was performed in four hospitals of Southern Poland (Krakow, Rabka, Limanowa, Zakopane). Lead content was determined in maternal and cord blood as well as in head and pubic hair by the GF AAS (Perkin Elmer). A significant interregional variation of lead content in maternal blood was observed. Lead concentration in maternal and cord blood was significantly higher in the group of mothers of SGA newborns when compared to the controls. This was not the case with respect to the mothers of preterm infants. Also, the comparison of lead concentration in head and pubic hair revealed no statistically significant case control differences. For a combined population of cases and controls, a significant gradient of lead concentration between maternal and cord blood was demonstrated. The correlation between lead content in different body compartments was observed. Conclusions. Different blood lead levels observed in mothers from four hospitals suggest different exposure. Higher lead concentration in maternal blood was associated with an increased risk of the delivery of SGA infant. PMID- 8711189 TI - The frequency of toxic methemoglobinemias in people living in the vicinity of refuse dumps in Barycz. AB - In the paper a thorough study of the influence of the methemoglobin levels and the occurrence of certain syndromes of clinical symptoms was made in the inhabitants living in the immediate vicinity of the large refuse dumps. During a 2.5-day clinical hospitalization the following examinations were done: general medical, specialistic and neuropsychological examinations, biochemical-analytical examination, EKG, chest X-ray, USG of the abdominal cave, spirometric tests, toxicological investigation of body fluids: carboxyhemoglobin, methemoglobin, lactates. The air pollution was measured and included: continuous measurement of the suspended particulate matter, carbon monoxide, sulfur dioxide, nitrogen oxide, nitrogen dioxide and the sum of nitrogen oxides. The evaluation of the refuse dumps gas showed that its main compounds were: carbon dioxide, methane in the amount of a few volume percent. Other pollutants were: carbon monoxide, hydrogen sulfide, methane homologues and aromatic hydrocarbons. The results of biochemical investigations were within normal limit. The elevated methemoglobin levels were found in 8 patients. The increased values of blood lactate concentration were found in 14 people and in 8 people the elevation of carboxyhemoglobin level was noted. The results of toxicological investigation do not unequivocally indicate the occurrence of toxic methemoglobinemia. However, the elevated levels of methemoglobin and lactates indicate a slight degree of oxygen deficiency in the body. In the examined people typical symptoms of methemoglobinemia were not observed, the only symptoms which may indicate their impact on the central nervous system were frequent headaches and neurovegetative disorders. PMID- 8711190 TI - The concentrations of nitrates and nitrites in food products and environment and the occurrence of acute toxic methemoglobinemias. AB - In the study it was intended to evaluate the concentration of nitrates and nitrites in some food products and samples of potable water collected in Krakow region and compile these results with frequency of recognised toxic methemoglobinemia cases. In the years 1991-1993 91 food samples, 288 samples of tap water from the central waterworks of Krakow and 399 samples of well-water from the country side of Krakow region were collected. In the chemical laboratories the nitrates and nitrites concentration has been defined. The clinical material of inpatients from 2 hospitals of Krakow and the data from the Poison Information Centre in Krakow have been analysed. The examined food samples did not present, in general, raised levels of nitrates and nitrites. Also the tap water samples presented a low concentration of these substances. The well-water samples demonstrated in considerable percentage increased levels of nitrates and nitrites and exceed the level permitted by sanitary law. In the Provincial Childrens Hospital during recent 14 years 8 infants with recognised methemoglobinemia were treated. All cases were connected with environmental pollution. At the Department of Toxicology, where only adults are treated, no methemoglobinemia cases resulting from environmental pollution during recent 26 years were hospitalized. The Poison Information Centre during recent 14 years consulted 239 cases of methemoglobinemia; 90% of them concerned children intoxicated because of environmental pollution. PMID- 8711191 TI - Does industrial environment influence the prevalence of arterial hypertension, plasma cholesterol and uric acid concentration and activity of the renin aldosterone system? AB - The present study aimed to assess the influence of a highly industrialized environment on the prevalence of arterial hypertension and plasma levels of cholesterol, uric acid, aldosterone and plasma renin activity. Epidemiological studies were performed in 5008 inhabitants of a highly industrialized city (Piekary Slaskie) and in 5004 inhabitants of non industrialized city (Olesno Slaskie) in the south part of Poland. Only subjects over 15 years were enrolled. The prevalence of arterial hypertension in the industrial population was 17% while in the country city 8.2%. Significantly higher serum cholesterol levels were found in hypertensive and normotensive subjects of the highly industrialized region than in the non industrialized one. Finally in hypertensive women aged < 40 years and normotensive men and women of the industrialized region significantly higher plasma renin activity and plasma aldosterone concentration were found than in corresponding population groups of the non industrialized region. From the results obtained in this study it seems, that the highly industrialized environment seems to stimulate hypertensinogenic factors. PMID- 8711192 TI - Ethylene glycol acute poisoning treatment results in Krakow in the years 1990 1994. AB - An analysis of ethylene glycol acute intoxication treatment results was performed in a group of 36 patients hospitalized within a five year period. Mean serum and urine glycol concentrations in the analyzed population ranged from 0-851 mg/dl (mean = 130 mg/dl) and from 12.4 to 930.0 mg/dl (mean 333 mg/dl), respectively. At the time of admission to the clinic 15 of 36 patients were deeply unconscious and mean acid-base balance values were as follows: pH 6.99, pCO2 16.7 mmHg, pO2 140.1 mmHg, HCO3 6.36 mmol/l, BE -29.6 mmol/l. Because of respiratory failure 21/36 patients (58.3%) required controlled ventilation and 24/36 (66.7%) underwent dialysis. Sixteen patients (44.4%) developed acute renal failure. Mean hospitalization period was 16 days (1-53). Eighteen patients (50%) died. The direct death mechanism in 15 patients (83.3%) was asystolia and in the remaining individuals other circulatory disturbances. The main reasons of high mortality rate were multiple organ damages secondary to severe metabolic acidosis. PMID- 8711193 TI - Long-term Cyclosporine A (Sandimmun) therapy for steroid resistant nephrotic syndrome in children. AB - The aim of the paper was to evaluate Cyclosporine A (CsA) treatment in children with steroid resistant nephrotic syndrome (diagnosed according to criteria established by ISKDC), in whom numerous attempts at other types of therapy had failed. CsA was used in 23 children aged 2-16 years. Renal biopsies revealed minimal changes (MCD) in 7 children, focal segmental glomerulosclerosis (FGS) in 11, mesangial glomerulonephritis (MES) in 4, and membrano-proliferative glomerulonephritis (MPGN) in 1. CsA was administered for a period of 6-41 months at a daily dose of 6 mg/kg, gradually decreased to 2.5 mg/kg. Blood CsA level was 40-200 ng/ml. In the course of the treatment, proteinuria disappeared or diminished and clinical improvement was seen in 21 patients. 13 children had relapses of nephrotic syndrome, showing very low blood CsA levels. Complete remissions were achieved in 16 children, partial remissions in 5, no remission in 2. Throughout the treatment and after its cessation 22 patients manifested no deterioration of renal function. In 1 child, repeated renal biopsies revealed some lesions that could suggest a nephrotoxic effect of CsA. Secondary steroid sensitivity observed in 3 children should be emphasized. Concluding, CsA is an effective and well tolerated agent in children with steroid resistant nephrotic syndrome, and the improvement noted in patients with FGS may reduce the risk of early terminal failure. The treatment requires monitoring blood CsA levels. PMID- 8711194 TI - Neurotoxicity of Cyclosporine A in therapy of steroid resistant nephrotic syndrome in a 10-year old boy. AB - A 10-year old boy with steroid resistant nephrotic syndrome developed neurological symptoms during the combined therapy with Cyclosporine A, prednisone and ketonazole. These symptoms might be caused by Cyclosporine because the clinical course and laboratory data revealed neither inflammatory nor other causative factors. PMID- 8711195 TI - The role of lead as an environmental pollutant in the period of growing ecological consciousness. AB - On the basis of the monitoring measurement results of Krakow's environment a long time evaluation has been presented, with regard to lead presence. The pollution with lead in a last few years was displayed in atmospheric air, soil and drinking water samples, as well as in some vegetables and food products. It was concluded that a characteristic trend regarding lead pollution of the environment could be observed; a decrease of lead level in air and in vegetables was presented, in soil and milk as well as in dairy products--remained low. Only in lettuce and parsley leaves the exceeding of the permissible lead content was confirmed. At present, in spite of the decrease of the heavy metals level in the environment, lead is still present and the health hazard to men exists, but to a lesser degree. PMID- 8711196 TI - Progressive hypomagnesemia in the industrialised part of southern Poland. PMID- 8711197 TI - The use of REMEDI HS in toxicological diagnostics of patients poisoned with drugs at the Department of Toxicology Collegium Medicum of the Jagiellonian University in Krakow. PMID- 8711198 TI - [Duties of an acute poisoning unit in a multiple profile provincial hospital]. PMID- 8711199 TI - [Toxic contamination of the rural environment in the province of Krak'ow and incidence of acute poisoning in humans. Clinical problems]. PMID- 8711200 TI - [State Fire Services in rescuing people injured in chemical accidents and disasters]. PMID- 8711201 TI - [Dangers connected with production and transport of toxic industrial chemicals (TSP) (summary)]. PMID- 8711202 TI - Free radicals, lipid peroxidation and sperm function. AB - The cellular generation of reactive oxygen species was first observed in mammalian spermatozoa in the late 1940s. The field then remained dormant for 30 years until Thaddeus Mann and Roy Jones published a series of landmark papers in the 1970s in which the importance of lipid peroxidation as a mechanism for damaging mammalian spermatozoa was first intimated. The subsequent demonstration that human spermatozoa produce reactive oxygen species and are susceptible to peroxidative damage has triggered intense interest in the role of oxidative stress in the aetiology of male infertility. Moreover, data have recently been obtained to indicate that, although excessive exposure to reactive oxygen species may be harmful to spermatozoa, in physiological amounts these molecules are of importance in the control of normal sperm function. This review considers the dualistic role of reactive oxygen species and sets out the current understanding of the importance of oxidative processes in both the physiology and the pathology of the human spermatozoon. PMID- 8711203 TI - Proceedings of the 7th International Symposium on Spermatology. Cairns, Queensland, Australia, 9-14 October 1994. PMID- 8711204 TI - Genetic control of mitosis, meiosis and cellular differentiation during mammalian spermatogenesis. AB - Gametogenesis in both the male and female mammal represents a specialized and highly regulated series of cell cycle events, involving both mitosis and meiosis as well as subsequent differentiation. Recent advances in our understanding of the genetic control of the eukaryotic cell cycle have underscored the evolutionarily-conserved nature of these regulatory processes. However, most of the data have been obtained from yeast model systems and mammalian cell lines. Furthermore, most of the observations focus on regulation of mitotic cell cycles. In the present paper: (i) aspects of gametogenesis in mammals that represent unique cell-cycle control points are highlighted; (ii) current knowledge on the regulation of the germ cell cycle, in the context of what is known in yeast and other model eukaryotic systems, is summarized; and (iii) strategies that can be used to identify additional cell cycle regulating genes are outlined. PMID- 8711205 TI - The azoospermia factor (AZF) of the human Y chromosome in Yq11: function and analysis in spermatogenesis. AB - Different Y mutations in Yq11 occurring de novo in sterile males were first described 19 years ago. Since the phenotype of the patients was always associated with azoospermia or severe oligospermia, it was postulated that these mutations interrupt a Y spermatogenesis locus in the euchromatic Y region (Yq11) called azoospermia factor (AZF). Recently, it became possible to map AZF mutations to different subregions in Yq11 by molecular deletion mapping. This indicated that azoospermia is possibly caused by more than one Y gene in Yq11 and the Yq11 chromatin structure. The frequency of AZF mutations in idiopathic sterile males (5-20%) may indicate a need for a general screening programme for its analysis in infertility clinics. PMID- 8711206 TI - 'Chauvinist genes' of male germ cells: gene expression during mouse spermatogenesis. PMID- 8711207 TI - A model for understanding gene regulation during spermatogenesis: the mouse testis Pdha-2 promoter. AB - Spermatogenesis is a complex process requiring the coordinate expression of a number of testis-specific genes. How these genes are regulated during spermatogenesis is poorly understood. However, the elucidation of these mechanisms has significant implications for both medicine and the primary livestock industry. The delineation of this process is of particular interest and, in this study, Pdha-2, a gene which codes for the murine testis-specific isoform of the E1 alpha subunit of the pyruvate dehydrogenase complex, has been used as a model. This review focuses on current knowledge about its expression and regulation during spermatogenesis. PMID- 8711208 TI - The molecular biology of SRY and its role in sex determination in mammals. AB - SRY is the gene that initiates the genetic cascade leading to testis development in mammals. Since its discovery in 1990 and the direct demonstration of its male determining role in transgenic mice, attention has turned to understanding the biochemical mode of action of the SRY gene product, and to the identification of other genes in the sex-determining pathway. Recent progress in these efforts is summarized in this review. PMID- 8711209 TI - Interleukin-1, interleukin-6 and the germ cell-Sertoli cell cross-talk. PMID- 8711210 TI - The epididymis as protector of maturing spermatozoa. PMID- 8711211 TI - The sperm centrosome during fertilization in mammals: implications for fertility and reproduction. AB - This article reviews the recent discoveries that: (1) nearly all mammals, including humans, inherit their centrosomes from their fathers; and (2) some sperm are ineffective in organizing the microtubules essential for effecting genomic union during fertilization, leading to the speculation that these sperm have centrosome defects. In addition, the molecular dissection and reconstitution of the human sperm centrosome in vitro is presented. PMID- 8711212 TI - Sperm competition: evolutionary causes and consequences. AB - The interaction between functional and mechanistic approaches to sperm competition and between male and female perspectives are described and illustrated by a study of the zebra finch, Taeniopygia guttata. Sperm competition experiments in the laboratory show that last male sperm precedence occurs (as it does in many other taxa) although the mechanism is unknown (as in most other taxa). Empirically-derived values were used to construct a mathematical model of sperm competition in the zebra finch. The model indicates that precedence occurs as a consequence of: (i) the temporal pattern of pair copulations; (ii) the rate at which sperm are lost from the female tract; and (iii) more sperm being transferred during extra-pair copulations than during pair copulations. The latter effect is a consequence of males seeking extra-pair copulations after their own pair copulation period has ended. The effect of sperm numbers on the pattern of sperm precedence may be further increased by: (i) extra-pair males increasing ejaculate size (sperm numbers) (for which there is no evidence); (ii) extra-pair males being of a better quality and transferring more sperm or better quality sperm (for which there is some evidence); and (iii) cryptic female choice. Females eject over 99% of sperm following insemination; if they eject fewer sperm from males chosen as extra-pair copulation partners, the potential for cryptic female choice is considerable. However, this is still being investigated. The model also predicts the optimal time for an extra-pair copulation to occur (from either a male or female perspective). A comparison between the predicted and observed pattern suggests that the optimal timing of extra-pair copulations is constrained in both sexes. PMID- 8711213 TI - Developmental expression and possible role of perinuclear theca proteins in mammalian spermatozoa. PMID- 8711214 TI - Three-probe fluorescence in situ hybridization to assess chromosome X, Y, and 8 aneuploidy in sperm of 14 men from two healthy groups: evidence for a paternal age effect on sperm aneuploidy. AB - The method of simultaneous three-chromosome fluorescence in situ hybridization (FISH) was developed using repetitive DNA sequence probes for chromosomes 8, X and Y and applied to semen of 14 men from two healthy groups who differed in their average ages (46.8 +/- 3.1 years, n = 4 v. 28.9 +/- 5.0 years, n = 10). The frequencies of disomic sperm determined by FISH compared well with frequencies obtained using the hamster-egg technique for human-sperm cytogenetics and with the frequencies of disomic and diploid sperm reported in previous FISH studies in this laboratory. The two groups of men did not differ in their baseline frequencies of sperm disomic for chromosome 8 (approximately 6.5 per 10(4) sperm), sperm with XY8 aneuploidy (approximately 9.5 per 10(4) sperm), or sperm with autodiploidy XX88 or YY88 (approximately 2 per 10(4) sperm). However, the older group had statistically higher frequencies of sperm carrying sex chromosomal disomy than the younger group (5.1 v. 2.2 per 10(4) sperm for XX8; 5.9 v. 2.0 per 10(4) sperm for YY8; P < 0.005). A recent report from this laboratory of sex-chromosomal aneuploidy in sperm of aged mice provides inter species corroborating evidence for this preliminary finding of a paternal age effect on sperm aneuploidy in human males. PMID- 8711215 TI - Spermatology for understanding, managing and conserving rare species. AB - Most conventional spermatology research involves common mammalian species including livestock, laboratory animals and humans. Yet, there are more than 4500 mammalian species inhabiting the planet for which little is known about basic reproductive biology, including sperm characteristics and function. This information is important, not just as adjunct knowledge, but because the majority of these species are threatened with extinction, largely due to human-induced pressures. The field of conservation is changing rapidly, and global cooperation is emerging among a variety of wildlife enthusiasts, ranging from management authorities of nature reserves to curators of rare zoological collections. Conservation progress depends on systematic, multidisciplinary research first to answer basic questions, with new data then applied to endangered species management plans. The reproductive physiologist is a crucial component of this scheme. Reproduction is the essence of species survival, and enormous effort needs to be directed at these 'untraditional' research species, subspecies and populations. Spermatology research combined with simultaneous efforts in endocrinology, embryology and cryopreservation (among others) can lead to the successful application of assisted reproduction. Examples from this laboratory include an array of wild felid species and a rare cervid and mustelid. Obstacles to success are formidable, including unique species-specificities, diminished genetic diversity and a general lack of resources. Nonetheless, the field offers tremendous opportunities for generating unique knowledge of comparative interest and with conservation utility. PMID- 8711216 TI - Sperm antigens and immunocontraception. AB - The development of novel forms of contraception is one way in which the world population crisis is being tackled. The concept of a contraceptive vaccine based on gamete-specific antigens is a particularly attractive approach. Much research has been carried out to identify sperm antigens which could be used as the immunogen. The most encouraging leads have come from groups using monoclonal antibodies to identify and characterize sperm antigens important for fertility (e.g. SP-10, PH-20 and PH-30). Identification of these molecules will also enable the development of specific tests for the diagnosis of immune infertility. PMID- 8711217 TI - Micro-assisted fertilization. AB - In couples who undergo routine in vitro fertilization (IVF), 17% experience significant problems with fertilization and many others are unable to have routine IVF because the quality of their semen is too poor. Often, the only options previously available to these couples were to use sperm donated by fertile men or to remain childless. Micromanipulative assisted fertilization techniques have improved the treatment of severe male factor infertility significantly and this paper provides a brief overview of the recent methodologies. Initially, techniques such as zona drilling and partial zona dissection, in which a hole or slit is placed in the zona pellucida, demonstrated that fertilization and pregnancies could be achieved with semen of very poor quality, but successes were sporadic. Later, subzonal injection of spermatozoa provided more consistent results with many units reporting pregnancies; however, relatively low rates of fertilization (14-34%) and high rates of polyspermy remained unresolved problems. The latest technique, the injection of a single spermatozoon into the oocyte cytoplasm, although technically difficult in animal models, proved to be highly successful in the human, restoring fertilization rates to those seen in routine IVF (65%) and producing good pregnancy rates from transferred embryos. Intracytoplasmic sperm injection has become the method of choice in the treatment of severe male factor infertility and preliminary data suggest that there is no increase in congenital abnormality among babies born after the transfer of injected oocytes. PMID- 8711218 TI - Use of epididymal sperm for assisted reproduction in men with acquired, irreparable obstructive azoospermia. AB - Microsurgical epididymal sperm aspiration (MESA) and in vitro fertilization (IVF) is primarily offered to men with congenital absence of the vas deferens (CAVD). However, the IVF capacity of these epididymal sperm is low ( < 15%) and unpredictable. In this study, IVF and intracytoplasmic sperm injection (ICSI) results in patients with non-congenital, irreparable obstructive azoospermia were analysed. Thirty-three patients were evaluated for a total of 37 cycles of MESA and IVF. Most had obstruction secondary to failed vasectomy reversal and to epididymal blockage. The overall fertilization rate was 30% with regular IVF and 26% with ICSI, and six clinical pregnancies were obtained. Both rates are significantly higher than the IVF rate previously reported with sperm from men with CAVD (13%, P < 0.00001). In men with non-congenital obstructive azoospermia, a significant difference was found in the average sperm count (56.9 x 10(6) v. 12.3 x 10(6), P < 0.04) and total motile count (16.6 x 10(6) v. 1.6 x 10(6), P < 0.01) respectively for patients who achieved IVF and those who did not. It is concluded that (a) the real IVF capacity of human epididymal sperm is 30%, or 42%, if calculated only for patients who achieved fertilization, (b) this higher rate is an indirect support of the hypothesis that sperm from men with CAVD have intrinsic biochemical defects, related to cystic fibrosis mutations, responsible for their low and unpredictable IVF rate, and (c) MESA and IVF can be offered at the same time or as an alternative to patients requesting vasectomy reversal. PMID- 8711219 TI - Regulation of sperm motility at the axonemal level. PMID- 8711220 TI - Derivation and reliability of kinematic measures of sperm motion. AB - Studies of sperm movement are relevant in the diagnosis of sperm function and in investigations of cellular biology. Such studies have been traditionally performed by analysing the kinematics of the flagellum or the head. Analysis of the flagellum can provide insights into the cell biological mechanisms responsible for the control of movement. However, the mathematical correspondence between head kinematics and flagellum kinematics is not unique. Therefore, it is not possible to use head kinematics to obtain detailed insights into cell mechanisms or physiology. The accuracy and precision of kinematic measurements are limited by a number of technical and biological factors. Therefore, the interpretation of kinematic data is dependent on a thorough understanding of the assumptions and conditions underlying the analysis. Evaluation of the reliability of kinematic measurements has suffered because no absolute standard for measurement has existed. The development and application of a new standard based on images which were simulated using the equations of motion is described. Because the kinematics of these images are known prior to empirical measurement, the performance of different methods can be determined absolutely. Some kinematic measures are unreliable because they are inappropriate analogues for engineering concepts. The development and use of appropriate engineering measures for the frequency and amplitude of sperm motion is also described. Some types of sperm motion cannot be analysed using kinematic measures (e.g. hyperactivated movement). The concept of the fractal dimension as a more accurate measurement for such motions is introduced. It is concluded that kinematic measurements of sperm motion can provide valuable information about cell biological mechanisms (in the case of the flagellum) and about general membrane and axoneme function (in the case of the head) when the measurements are made under the appropriate conditions, when standard techniques are followed, and when the assumptions underlying the analysis are well understood. PMID- 8711222 TI - Sex preselection by flow cytometric separation of X and Y chromosome-bearing sperm based on DNA difference: a review. AB - Recent research on the flow cytometry of sperm for the purpose of predetermining gender of offspring has led to a validated method to separate X from Y chromosome bearing sperm for use with in vitro fertilization and embryo transfer, intratubal insemination or intracytoplasmic sperm injection. The basis for the method is the sex chromosome-specific marker, DNA, which is present in greater amounts in X bearing sperm than in Y-bearing sperm of mammals. Sperm are exposed to the vital dye Hoechst 33342 which binds to the minor groove of the DNA helix. Flow cytometric sorting of the sperm using a laser as the excitation source results in populations of Y- or X-bearing sperm that are 85-90% pure. Several hundred offspring have been produced from swine, rabbits, sheep and cattle that confirm the predicted sex. The method is currently being applied to the commercial embryo market. The method is not likely to be used in conjunction with standard cattle or swine artificial insemination practice in its current form since only about 4 x 10(5) sorted sperm can be produced per hour of sorting. The technology has also been applied to human sperm for use by couples that are at risk to sex-linked disease expression in their offspring. Populations of human sperm have been sorted with X and Y purities of about 80% as confirmed by DNA probe technology and fluorescence in situ hybridization. PMID- 8711221 TI - Recent developments and concepts in the cryopreservation of spermatozoa and the assessment of their post-thawing function. AB - New research on the cooling and cryopreservation of mammalian spermatozoa is reviewed in the context of the older literature. Cryoinjury to a variety of cell organelles is regarded as being due to the two major stresses of cryopreservation, i.e., the change in temperature, and the formation and dissolution of ice and its consequences. Since the cryopreservation process involves departure of the cells from and return to body temperature, both cold shock and warm shock are included as potential stresses to be considered, as well as the stages involving cooling below the freezing point of the medium. The causes of cryoinjury are reconsidered and new concepts concerning the influence of osmotic stress are presented. Heterogeneity of the sperm population is discussed in the context of the success with which spermatozoa can be cryopreserved between and within ejaculates and individuals. The functional state of frozen and thawed spermatozoa is examined on the basis of published results of structural and functional tests of sperm competence. The hypothesis is advanced that cryopreserved mammalian spermatozoa are in a state resembling capacitation, which accounts for their relatively reduced longevity and their readiness to undergo egg penetration without incubation. The importance of this to the utilization of cryopreserved spermatozoa is examined, and proposals are made for new avenues of research to overcome these problems. PMID- 8711223 TI - Ionic control of sperm function. AB - Successful sperm function leads to fertilization. It is dependent on the extracellular environment, especially the array and concentration of various ions. Considerable evidence indicates that this is because of consequent effects on the intracellular ionic composition. Although both cations and anions undoubtedly play a role in a modulating sperm function, most of the evidence currently available concerns cations. Therefore, this review will concentrate on cations, focussing on Ca2+, Na+, K+ and H+. Their requirements for successful capacitation (mammalian sperm) and acrosomal exocytosis (both invertebrate and mammalian sperm) will be considered. In particular, the mechanisms which may control ion fluxes, leading to changes in the intracellular ionic composition and subsequently to changes in sperm functional potential, will be addressed. PMID- 8711224 TI - Interactions between gametes leading to fertilization: the sperm's eye view. AB - Sexual reproduction requires that the gamete carrying the male-derived haploid chromatin join with the gamete carrying the female-derived haploid chromatin during fertilization to produce the diploid zygote. To accomplish this feat, the sperm must not only meet the egg, it must recognize the egg and be recognized in turn by the egg, and in the end must enter and be engulfed by the egg. In this selective overview of gamete interactions that lead to fertilization, encounters of three kinds, followed by the finale of gamete fusion, are considered from the sperm's viewpoint, with particular emphasis on the mammalian species with the mouse as the principal model. The first encounter is with the zona pellucida of the egg, to whose surface the sperm must bind. Mouse sperm appear to have four binding sites for zona ligands. Three interact with sugar moieties of the oligosaccharide chains of the mouse zona glycoprotein ZP3; the fourth binds a peptide backbone arginine. Capacitation is not required for this encounter, but is obligate for the second encounter--induction of the acrosome reaction in the bound sperm. The acrosome reaction is an exocytotic process that makes available the enzymatic machinery needed for sperm penetration the zona which is the end point of a sequence of reactions directed by intracellular signalling systems. In mouse sperm, these systems are presumed to be activated by ligands on ZP3 binding to ligand-specific sperm receptors with consequent aggregation of receptors. No receptor has been identified with certainty, nor have candidates for putative ZP3 ligands been identified. Completion of the acrosome reaction allows the sperm to penetrate the zona and, bind to the egg plasma membrane, thereby completing the third encounter. In the mouse, a 94-kDa protein appears essential for this binding. In the guinea-pig, a sperm plasma membrane protein (formerly PH-30, now fertilin), is a strong candidate for the mediator of the fusion process by which the egg engulfs the sperm. Decondensation of the sperm chromatin reverses the remarkable packing of DNA organized by sperm protamines. Mitochondrial DNA is also engulfed by the egg; the question of whether this DNA makes a small finite, or null, contribution to cytosolic inheritance is still in debate. The puzzles attending these encounters are presented as reminders of the intricacy and fascination, as well as of the vital necessity, of gamete interaction. PMID- 8711225 TI - Synchronous multispecific spawning on coral reefs: potential for hybridization and roles of gamete recognition. AB - Problems of gamete recognition in corals and other mass-spawning invertebrates are potentially great. In 'mass spawnings', closely-related species or genera commonly spawn at the same time, or within 1 or 2 h of each other, increasing the potential for hybridization. Among mass-spawning corals, most of the species involved are hermaphrodites that package the gametes in buoyant bundles that float to the sea surface before breaking up. Local hydrodynamic features frequently act to aggregate gametes from many different species into slicks where both eggs and sperm can be viable for extended periods. Other mass-spawning invertebrate taxa, such as molluscs, polychaetes and various echinoderms, do not have buoyant gametes but also spawn with a high level of synchrony. Gametes of organisms participating in these spawning events must be able to successfully recognize conspecifics. If they cannot do this, either through sperm chemotaxis or by mechanisms at the level of sperm binding and penetration, there may be high levels of gamete wastage through hybridization. Alternatively, viable hybrids may be formed, a factor that could have contributed to the evolutionary history of mass-spawning taxa, as well as to the taxonomic difficulties that have plagued the taxonomy of groups such as reef-building corals. Within some mass-spawning taxa, pre-zygotic barriers to fertilization suggest relatively recent molecular evolution at gamete recognition loci. PMID- 8711226 TI - Workshop report: clinical CASA--the quest for consensus. AB - The goals of the Workshop were to establish the current state-of-the-art in the clinical use of computer-aided sperm analysis (CASA), to identify areas of contention or confusion in the use of CASA technology, and to reach consensus on these matters to facilitate the wider use, and usefulness, of these instruments in clinical and research applications. CASA assessments of sperm morphology were not considered as they were discussed in a separate Workshop (Advanced Techniques in Sperm Preparation and Imaging) on analytical techniques. Four topics were considered: (a) CASA and semen analysis; (b) the diagnostic value of sperm kinematics in semen; (c) human sperm hyperactivation; and (d) CASA and fertility prediction. In all, 17 specific consensus points were identified. PMID- 8711227 TI - [Cotrimoxazole induced dermatitis and curative treatment of AIDS pneumocystosis]. AB - Adverse cutaneous reactions frequently occur during the treatment of AIDS associated pneumocystosis by trimethoprime-sulfamethoxazole. The most common form is a maculous rash. Treating throughout the duration of hypersensitivity may lead to potentially lethal Stevens-Johnson and Lyell syndromes. Slow acetylator phenotype, a glutathion deficiency and a history of adverse cutaneous reactions have been identified as risk factors of cutaneous reactions. An adjuvant corticosteroid therapy decreases the frequency of adverse cutaneous reactions during the treatment of hypoxaemic pneumocystosis by trimethoprim sulfamethoxazole. PMID- 8711228 TI - [Respiratory mechanics of chronic obstructive lung disease in acute respiratory insufficiency]. AB - Respiratory mechanics abnormalities in patients with chronic obstructive pulmonary disease (COPD) in acute respiratory failure (ARF) consist of the followings : 1) expiratory flow limitation, 2) marked increase in airway resistance, 3) dynamic hyperinflation. As a results, both resistive and elastic loads to the respiratory muscles are increased. These abnormalities, which are already present in stable COPD patients, are considerably more marked in ARF. Our contribution was to systematically describe the passive mechanical properties of lung and chest wall of COPD patients tracheally intubated, mechanically ventilated, sedated-paralyzed for ARF. Mechanical properties, i.e. resistances and elastances, were obtained from the rapid end-inspiratory airway occlusion technique during constant-flow inflation. This method allows to partition the resistance into its two components, namely the interrupter resistance, which reflects airway resistance, and additional tissue resistance, which pertains to time constant unequalities and/or viscoelastic behavior. We also determined the static and dynamic elastances of both lung and chest wall. Static intrinsic positive end-expiratory pressure (PEEPi) was obtained from end-expiratory airway occlusion. By changing, for one breath, inflation flow, at constant volume, and inflation volume, at constant flow, we investigated the time, and hence the frequency dependence of resistance and elastance. In addition, we divided the inspiratory work of breathing into its four components which are the PEEPi component, the static work, the purely resistive work and the additional work. Finally we compared our results with those of normal anesthetized and paralyzed subjects. We found that airway resistance was markedly higher in COPD, as were also the additional resistance and the dynamic elastance of the lung. Additional resistance and dynamic elastance of lung and chest wall exhibited a marked frequency dependence in COPD. Shortening the inspiratory time could result not only to reduce the hyperinflation but also to increase expiratory flow through the increased dynamic pulmonary elastance. The inspiratory work was twice higher in COPD than in normals because of the PEEPi and the resistive components. Due to their flow and volume dependence, the results of resistances and elastances should be standardized. PMID- 8711229 TI - [Cellular growth factors]. PMID- 8711230 TI - [Cellular growth factors and cancer]. PMID- 8711231 TI - [Physiopathologic mechanisms of drug-induced lung diseases in man]. AB - Iatrogenic lung disease in man is generated by very different and often complex pathology. This explains the great clinical diversity of these disorders which may manifest as eosinophilic pneumonia, intra-alveolar haemorrhage, bronchiolitis obliterans and diffuse interstitial pneumonia even with pulmonary fibrosis. The causes are also very varied such as direct cellular toxicity, cellular oedema, an alteration of the alveolar capillary membrane, the activation of inflammatory and/or immune cells, which are responsible for the production of soluble mediators whose effects are sometimes harmful to the pulmonary parenchyma. Rather than reporting on the different clinical types of iatrogenic lung disease and indicating for each one the hypothetical or known physiopathogenic mechanism, we have chosen to examine certain fundamental lesional mechanisms and to indicate the principal nosological groups which they cover. We have centered this review on the physiopathogenic models which are the most coherent and most fully elaborated based on observations made on man or on experimental animal models. Among those we have reported here is a case of bleomycin toxicity, with its direct toxic mechanism on the epithelial or endothelial cellular targets, amiodarone lung disease and with its associated alveolar oedema, inflammatory reactions and immunological reactions whose specificity is poorly understood; also there are some alveolitides whose specificity has been demonstrated, such as those to minocycline and to BCG and finally a complex model which is both inflammatory and disturbed immunology in radiation pneumonia. PMID- 8711232 TI - [Iatrogenic pulmonary artery hypertension]. AB - Primary pulmonary arterial hypertension (PPH) is a rare disorder with a predilection for young subjects (most commonly of 20-40 years) and of the female sex. The prognosis is very poor because the average life expectancy is of the order of two to three years from the time of diagnosis. Since the epidemic of PPH observed in Switzerland, Austria and Germany in the years between 1968 and 1970, following the arrival of a derivative of amphetamine, Aminorex, it was realised that certain medications, notably appetite suppressants, might play an initiating role. More recently, fenfluramine and dexfenfluramine have been incriminated: in a significant statistic of A-beclere's team, around 20% of cases of PPH were observed in patients who had taken fenfluramine or dexfenfluranime for more than three months. A very recent case-control study has shown that any use of anorexic drugs (mainly fenfluramine derivatives) was associated with an increased risk of PPH (odds ratio: 6/1), and particularly when the duration of treatment exceeds 3 months. Outside the group of appetite suppressants few medications are capable of favouring the development of PPH, the cases are sporadic and the relationship between cause and effect is hardly established. PMID- 8711233 TI - [Spiral CT angiography in the diagnosis of central pulmonary embolism: comparison with pulmonary angiography and scintigraphy]. AB - PURPOSE: To evaluate the accuracy of spiral CT angiography in the diagnosis of central pulmonary embolism (PE). MATERIAL AND METHODS: A retrospective study was undertaken in a population of 90 patients who were evaluated with spiral CT angiography (acquisition: 12 to 24 s with or without strict apnea; injection of 90 cm3 of 12, 20 or 30% contrast material at a rate of 4 to 7 cm3, selective pulmonary angiography of each lung (n = 55) and/or ventilation-perfusion (VP) scanning (n = 35). RESULTS: Among the 55 patients evaluated with both spiral CT and angiography, central pulmonary embolism was excluded in 19 patients (34%), assessed in 29 patients (53%) whereas CT examination was considered as inconclusive in 7 patients (13%) due to interpretive difficulties at the level of obliquely oriented arteries and/or presence of hilar lymph nodes (sensitivity: 90.5%; specificity: 82.6%). With spiral CT, the finding of 95 central emboli (3 main, 61 lobar and 31 segmental) corresponded exactly to the angiographic findings; spiral CT enabled direct visualization of intraluminal filling defects (n = 95) whereas the angiographic recognition of PE was based on direct (n = 57) and indirect (n = 38) signs. Spiral CT angiography was more sensitive and specific than VP scanning. CONCLUSION: Spiral CT angiography appears as a sensitive and specific noninvasive method for the diagnosis of central PE. PMID- 8711234 TI - [Comparative study of the performance and ergonomics of nebulizers in cystic fibrosis]. AB - This study had, as its aim, to test twelve nebulizers (6 jet, 6 ultrasonic) which are used in the treatment of cystic fibrosis. Devices were connected to a respirator in order to mimic the ventilation of a child and of an adult suffering from cystis fibrosis. Three medications: tobramycine, colistine and amiloride were nebulised. The volume of the recommended solution varied between 1.5 and 13 ml according to the manufacturer. During a session of ten minutes the ultrasonic nebulizer delivered an inhaled volume which was significantly greater than the jet (2.72 +/- 0.98 ml vs 1.22 +/- 0.59 ml, p < 0.0001) for the three drugs. Regarding granulometry, the fraction of particles between 0.5 and 5 microns, was higher with ultrasonic than with pneumatic nebulizer for tobramycine (67.1 +/- 10.7 vs 55.5 +/- 11.5%, p < 0.001) and amiloride (66.4 +/- 9.2% vs 58.1 +/- 15%, p < 0.05%). The variation of concentration due to nebulisation were independent of the type of apparatus but influenced by the drug since concentration was increased for tobramycine (+10.5 +/- 18.6%) and amiloride (+13.4 +/- 8/9%). In summary the effective fraction resulting from the inhalable fraction, from granulometry and from changes in concentration was significantly greater for ultrasonic than for jet nebrulizer (17.3 +/- 6.7% vs 9.7 +/- 9.6%, p < 0.001). This study underlines the great variability of the performance of aerosols generators and therefore the need for an accurate evaluation of nebulizer performances in order to prescribe the best nebulizer/drug association in clinical practice. PMID- 8711235 TI - [Tracheotomy: review of five years at the University Hospital of Lome, Togo]. AB - From a review of 37 cases, this retrospective study relates the activity of the Otorhinolaryngology service at the University Hospital Center of Lome, as regards tracheostomy. It details: - a high rate of urgent tracheostomy (76%) for laryngeal carcinoma and papillomatosis; - a high rate of complication (29%) and death (5%). This is due to the urgency of tracheostomies, unsuitable cannulae, irregular nursing and prolonged cannula carrying. A better technique and rigorous supervision could reduce these complications. PMID- 8711236 TI - [Asthma attacks in children over two years old in a pediatric hospital in Nice]. AB - Asthma is a common pediatric emergency. We report the results of a prospective investigation concerning asthma attacks observed in children aged more than 2 years at Lenval hospital (Nice) during a 12-month period, from September 1993 to September 1994. One hundred and thirty seven asthma attacks were observed in 122 children aged 6.9 +/- 3.4 years, representing 5.2% of children ranged from 2 to 6 years medical emergencies, and 10.3% of children greater than 6 years medical emergencies. In most cases (82.4%) the diagnosis of asthma was already known. The children were first seen, on the average, between 32.3 +/- 38.6 hours after the beginning of the attack, previously self-treated at home in one out of every two cases. Treatment was set in each case on the isolated administration of inhaled beta 2-adrenergic agonist. The decision whether or not to hospitalize was made, at the latest, one half hour after the second administration of the beta 2 agonist. One death occurred before hospital admission. Hospitalization was decided in 38.6% of the cases, two of which were immediately directed for reanimation. A majority of the children (60.7%) were able to return home after metered dose inhalation for 5 of them, after 1 nebulization of beta 2-agonist in 66 cases, after 2 nebulizations in 12 cases. The prevalence of asthma is increasing and can be estimated at approximately 10% in France. The pediatrics emergency wards have an important role to play in the evolution of this illness. They must guarantee the most favorable management of the attack. They must then participate decisively in the initiation of the necessary surveillance to be continued. They play a great part in the knowledge of the epidemiology. PMID- 8711237 TI - [Acute community-acquired pneumonia of moderate and grave severity investigated by bronchoscopy. Analysis of 193 cases hospitalized in a general hospital]. AB - Between February 1989 and June 1994 193 cases of acute community acquired pneumonia (PAC) which were of intermediate or great severity were admitted to two hospitals in the South West of France. These patients were explored using bronchofibroscopy (FB) with a protected brush (BP) and alveolar microlavage (MLBA) and quantitative cultures were performed, also there were other specimens taken in a regular fashion. The percentage of positive examinations was 60% for brushings (BP), 59% for MLBA and 21% for blood cultures and 16% for serological tests. An aetiology was determined in 137 cases (70.9%). The organisms recovered were Streptococcus pneumoniae (49.6%), gram negative bacilli (17.4%), Haemophilus influenzae (11.7%), Mycoplasma pneumoniae (4.4%), Mycobacterium tuberculosis (4.4%), Staphylococcus aureus (3.6%), Chlamydia pneumoniae (2.2%), Legionella pneumophila (0.7%), and various 5.8%. The overall mortality was 15% despite immediate antibiotics based on the likely organism in 88% of cases. The study of prognostic factors confirmed the Fine score system (determined a posteriori) which constitutes a useful and practical index determining the management of PAC. On the other hand the role of bacteriological documentation in improving the vital prognosis remains to be confirmed. If bronchofibroscopy has appeared to us as a safe and useful means of investigation, the management of these disease remains to specified. We suggest that its use is reserved for subjects with life threatening disease (a Fine score equal to or greater than 3) or for those patients who are likely to have unusual germs: failure of previous antibiotics, diabetes, malnourishment, cancer, airflow obstruction and inhalation. PMID- 8711238 TI - [Pulmonary metastases with a prolonged development. Two cases of thyroid cancers]. AB - The authors report two cases of differentiated carcinoma of the thyroid with pulmonary involvement showing as miliary shadowing radiologically which preceded the diagnosis of the thyroid neoplasm by 35 and 6 years respectively. The two patients had undergone cervical radiotherapy in infancy for lymphadenopathy whose aetiology had not been determined. The scintigraph with iodine 131 showed tht in two cases there was a bilateral and diffuse pulmonary uptake in keeping with pulmonary lesion of metastatic origin. Our observations recall the possibility of a slow evolution of pulmonary metastases and carcinoma of the thyroid and the role of cervical irradiation in the development of such cancers. With miliary shadowing a metastatic origin, in particular that of the thyroid, should be considered and in the majority of cases the proof could be supported using iodine 131 scintigraphy. The delay in appearance of pulmonary metastases during the course of cancer of the thyroid is variable. They occur most often after the initial diagnosis or more rarely preceding the discovery of a primary thyroid cancer. The pulmonary metastases may be asymptomatic. This neoplasm may benefit from a specific effective therapy and prolonged remissions have been described even with metastases. PMID- 8711239 TI - [Carcinoid thymus tumor at an advanced age: diagnostic value of mediastinal needle biopsy with computerized tomography]. AB - Carcinoid tumour of the thymus is a rare neuroendocrine tumour particularly at an advanced age. The authors report a case of a mediastinal mass in a man aged 85, the mass had remained asymptomatic for a long time. It was decided to achieve a diagnosis because the tumour was causing local compression: a mediastinal needle biopsy under computerised tomographic control confirmed that this was a carcinoid tumour and a study of the biopsy material using an electron microscope showed neurosecretory granules. A sternotomy enabled the tumour to be excised but a post operative Pseudomonas pneumonia led to the death of the patient. This case underlines the diagnostic place of mediastinal needle biopsy in the presence of a mediastinal tumour. The technique can be carried out under computerised tomography or ultrasonography and this can be associated with a study of the biopsy specimen using electron microscopy which enables the diagnosis to be made before any therapeutic decisions. The treatment of choice of a carcinoid tumour of the thymus is surgery which confirms the tumour limits and also its thymic origin. Tumour excision can be completed using radiotherapy or even chemotherapy. PMID- 8711240 TI - [Rhinomanometry]. PMID- 8711241 TI - [Vitamin A derivatives and prevention of bronchial cancers]. PMID- 8711242 TI - [Bibliometry of biomedical periodicals. Addendum]. PMID- 8711243 TI - [Current data on diagnostic imaging methods in acute pulmonary embolus]. PMID- 8711244 TI - [Complications of tracheotomy]. PMID- 8711245 TI - [Growth factors. From the laboratory to the clinic]. PMID- 8711246 TI - [Exercise training: its value in chronic obstructive lung disease]. AB - This review deals with the physiopathological background that justifies training techniques for COPD patients. Personal results are analysed in regard with the principal articles related to this field. PMID- 8711247 TI - [Fifty years of the journal Recent Advances in Medicine]. PMID- 8711248 TI - [Evaluation of two treatment schedules with different doses of interferon in chronic HBsAg+ anti-HBe+ hepatitis]. AB - Twenty-three patients with hepatitis B were treated with recombinant interferon alpha-2a. Twelve patients were given 6 MU three times a week for 6 months (Group A); eleven patients were given 9 MU three times a week for an initial 4-month course and subsequently for a further 3-month course after 6 months of no therapy (group B). Eight patients in the group A (66.7%) and seven patients in the group B became negative for HBV-DNA and normalized their ALT, compared to 31% of controls. Relapses were noticed in 75% of the responders in the group A and in 71.4% of the responders in the group B within 12 months after interferon discontinuation. The Authors are consequently of opinion that it is appropriate to try longer treatments with moderate doses of interferon in order to maintain the response as long as possible. PMID- 8711250 TI - [The Italian Cochrane Center and finding randomized clinical trials published in Italian journals: a European project]. PMID- 8711249 TI - Prognostic value of some laboratory parameters in a group of intravenous drug users in the early stages of HIV infection. AB - During a 5 year follow-up we found significantly reduced survival for non accidental deaths in 37 intravenous drug users (IVUDs) in the early stages of HIV infection when compared with 32 HIV-negative IVUDs (p = 0.017). Moreover, in HIV positive subjects, survival was significantly reduced for those groups which at the beginning of the follow-up showed the following values: circulating CD4+ lymphocytes <250/mmc (p = 0.007), CD4+/CD8+ ratio < 0.5 (p = 0.027), serum albumin < 4.13 g/dl (p = 0.045), IgA > or = 2.5 g/l (p = 0.043), IgM < 1.8 g/L (p = 0.041) and platelet count < 130 x 1,000/mmc (p = 0.038). In HIV positive patients, the value of 250 units/mmc for the circulating CD4+ lymphocytes still remained the most predictive parameter of increased mortality for disease at 5 years, even following investigations conducted with other cut-offs. Relationships existing at the beginning of the follow-up between circulating CD4+ lymphocytes and other prognostic parameters suggest that IgA (r = -0.34; p = 0.04), serum albumin (r = 0.33; p = 0.05), and CD4+/CD8+ ratio (r = 0.72; p = 0.0001), but not IgM (r = 0.25; N.S.) and platelets (r = 0.07; N.S.), are dependent variables of shortened survival. PMID- 8711251 TI - [Occult gastrointestinal hemorrhage. Description of a case with unusual clinical presentation]. AB - Bleeding from small bowel is a quite rare event and often is a diagnostic challenge to physician and surgeon. We present a case of a patient with an acute massive haemorrhage due to jejunal diverticulosis and with an unusual clinical setting. The site of bleeding was localized by scan with radiotagged erythrocytes, but the diagnosis of jejunal diverticule was evident only at laparotomy. The patient underwent to surgical resection of the affected bowel (40 cm). Although jejunal diverticula are considered a rare source of gastrointestinal haemorrhage, we suggest that this disorder must be considered in all patients with occult gastrointestinal bleeding especially in the elderly. PMID- 8711252 TI - [New perspectives in the treatment of hypertension]. PMID- 8711253 TI - [Drug-induced asthma]. PMID- 8711254 TI - [Oral immunotherapy in oculorhinitis from Gramineae]. AB - We have evaluated the efficacy of oral immunotherapy in 34 patients affected by seasonal allergic rhinoconjunctivitis due to grass pollen. Diagnosis was performed on the basis of medical history, Skin Prick Test and Rast and the mean duration of the disease was 6 years. The treatment started in November and consisted of increasing daily doses of allergen; when the maximum dose was reached, it was continued twice a week till April. After two years of treatment we observed a statistically significant reduction in the severity of the symptoms considered and in the amount of oral antihistamines taken during the pollen season. The treatment was well tolerated and patients did not experienced any adverse effect. We therefore conclude that oral immunotherapy is a valid alternative to traditional subcutaneous immunotherapy in the treatment of seasonal allergic rhinoconjunctivitis due to grass pollen. PMID- 8711255 TI - [Alpha interferon in the therapy of polycythemia vera]. AB - In previous researches recombinant interferon alpha (IFN-alpha) has been demonstrated to significantly control red cell mass and thrombocytemia in patients with polycythemia vera (PV). Further evaluation of drug effectiveness and of modalities of maintenance therapy is warranted. We treated four patients with PV according to PVSG criteria with IFN-alpha (3 MU subcutaneously three times a week) for five months. Thereafter the starting dose was reduced to 1.5 MU three times a week. Treatment with IFN-alpha at the higher dosage induced regression in sizes of the spleen and a return to normal levels of peripheral blood platelets and leukocytes. Phlebotomies, previously performed to keep under control hematocrit values, were no more needed. During maintenance treatment with IFN-alpha reduced dose platelet level remained in the normal range, spleen size did not show further variation but hematocrit slowly rose and phlebotomies had to be resumed. These results confirm IFN-alpha effectiveness in PV, but suggest the need of relatively high dosages of the drug and difficulties in switching to a maintenance treatment. PMID- 8711256 TI - Heeding the challenges. PMID- 8711257 TI - Sexuality and sexual capacities of elderly people. AB - Sexual behavior depends on biological and psychological factors. A gradual decline in interest and capacity may occur with advancing age. Sexual relationships have patterns that are influenced by prior degrees of sexual tension and outlet, physical and emotional status, and the mutual needs of the partners. In this article, the authors explore sexuality, psychosocial issues, and sexual capacities of elderly people, including issues of menopause and erectile dysfunction. The role of rehabilitation nurses in helping older adults understand and adjust to changes is addressed. Sexual counseling, already a part of rehabilitation nursing for younger patients, also should be provided for elderly patients. Nursing goals and strategies need to address identifying sexual problems and educating elderly rehabilitation patients about altered sexual needs and capacities. To do this, rehabilitation nurses must be knowledgeable about the physiology and sexual needs of older adults, be aware of myths about sexuality, and understand their own values and attitudes regarding sex and sexuality as well as the values and attitudes of individual patients. PMID- 8711258 TI - Family caregiving in cases of hip fracture. AB - The growing incidence and cost and the serious nature of hip fracture in the elderly require a closer examination of the family's role in the rehabilitation process and how it can be assisted in that role. In this prospective study, 57 family caregivers provided information before the hospital discharge of the patient with hip fracture and at 2, 8, and 14 weeks postdischarge. They were asked about caregiving demands and problems, caregiver mood, expectations about recovery, and advice to future caregivers. A brief follow-up was conducted at 6 months. The care recipients were all women and had been living at home before injury. The postdischarge location of the patient (e.g., in a residence shared with the caregiver, in a different residence, or in a nursing home) was a major factor in the types of caregiving activities but not in total demand. Nonspouses cited the most problems. Mood distress did not change over time. Caregivers appeared to have unrealistic expectations about the length of the recovery period; by 14 weeks, 35% judged the care recipient's mobility to be worse than expected, and 20% felt that the patient had more pain than expected. The most frequent advice to future caregivers was to have patience and to give encouragement. PMID- 8711259 TI - An interdisciplinary clinical advancement program within a patient-centered care model. AB - Restructuring in health care does not have to compromise the pursuit of clinical excellence and quality patient care. The clinical advancement program (CAP) at the Hospital for Special Care is a newly developed multidisciplinary reward and recognition program for clinical staff. The program is integrated into the hospital's structure of service line management and, unlike traditional advancement programs, is open to all levels of care providers: professional personnel, technical staff, and aides. This article describes the basic features of the CAP model and how its was developed by a multidisciplinary task force. PMID- 8711260 TI - "Aloha": an in-house nursing home vacation. AB - This article describes a weeklong in-house "vacation" that was held at a Department of Veterans Affairs nursing home. The author provides the guidelines used to plan, implement, and evaluate the project. The in-house nursing home vacation was successful, as measured by the number of residents in attendance, their positive evaluations, and their desire to repeat the experience. The primary reason for the success of this program was the number and strength of the interactive partnerships. PMID- 8711261 TI - Geriatric rehabilitation: nursing interventions and outcomes focusing on urinary function and knowledge of medications. AB - This article describes the nursing interventions that focus on urinary function and medication knowledge of older adults in a geriatric rehabilitation program. The authors review the interventions needed to achieve bladder retraining and self-administration of medication and consider what predicts urinary function and knowledge of medication administration in older adults. The descriptive study included 251 consecutive patients who were admitted into a geriatric rehabilitation unit. An evaluation of urinary function was done on admission and based on patient need to (a) maintain urinary function, (b) decrease urinary incontinence, and (c) resolve urinary retention. All patients received routine medication teaching from admission to discharge during each daytime administration of medication. From admission to discharge, there was a decrease in incidences of urinary incontinence and urinary retention and an increase in knowledge of medication regimens. Although the study was descriptive, the findings suggested that nursing interventions in geriatric rehabilitation may decrease urinary incontinence and retention and improve knowledge of medication regimens in this population. PMID- 8711262 TI - Rehabilitation and dialysis: a collaborative program. AB - Persons with end-stage renal disease who receive dialysis (hemodialysis or peritoneal dialysis) are candidates for rehabilitation when stroke, amputation, or debilitation affects their ability to function independently. A collaborative program between the rehabilitation and dialysis departments was developed to enable dialysis clients to participate fully in a rehabilitation program while they undergo dialysis treatments. Clients receiving hemodialysis are treated in the dialysis department three times week, while peritoneal dialysis clients are treated on the nursing unit. All dialysis clients receive the same therapy program as other rehabilitation clients do, with modifications as necessary. Hemodialysis clients may receive therapy during dialysis as deemed appropriate by the interdisciplinary team, which includes the dialysis nurses. PMID- 8711263 TI - Brian's world. PMID- 8711264 TI - No place like home. PMID- 8711266 TI - Effect of rh-GMCSF and rh-GCSF on oxygen free radical production by human neutrophils and blood monocyte-derived human macrophages. AB - The in vitro effect of recombinant human granulocyte-macrophage colony stimulating factor (rh-GMCSF) and recombinant human granulocyte colony stimulating factor (rh-GCSF) on oxygen free radical (OFR) generation by human neutrophils and blood monocytes derived human macrophages stimulated with phorbol myristate acetate was investigated and compared. The production of OFR by neutrophils and macrophages was time dependent, and the maximum release of OFR by neutrophils and macrophages was measured 90 and 180 min after stimulation with phorbol myristate acetate, respectively. The priming effects or rh-GMCSF and rh GCSF on OFR production by human neutrophils and macrophages was dose dependent. The maximum generation of OFR by neutrophils occurred when primed with 1,000 U/ml of rh-GMCSF and reached 2.383 +/- 0.191 nmol/10(5) neutrophils/90 min as compared with 1.072 +/- 0.113 nmol/10(5) neutrophils/90 min in the unprimed controls. This represents a 122.20% increase in OFR generation (p < 0.001). However, the percentage of maximum increase in OFR production was 57.84 when neutrophils were primed with a concentration of 5,000 U of rh-GCSF/ml. In 72-hour-old human macrophages, much higher levels of OFR production as compared with neutrophils were measured following stimulation with phorbol myristate acetate. The maximum generation of OFR was measured in macrophages primed for 45 min with 500 U/ml of rh-GMCSF. These cells produced 8.960 +/- 2.075 nmol/5 x 10(4) macrophage/180 min as compared with 4.563 +/- 1.773 nmol/5 x 10(4) unprimed macrophages/180 min (p < 0.001). In macrophages primed with rh-GCSF, however, the maximum OFR production was induced by a dose of 5,000 U/ml and reached 6.902 +/- 1.463 nmol/5 x 10(4) macrophages/180 min as compared with 4.563 +/- 1.773 nmol/5 x 10(4) macrophages/180 min in the unprimed controls (p < 0.05). In conclusion, the priming effect of rh-GMCSF on OFR generation by human macrophages and neutrophils was more potent than that of rh-GCSF, both in the extent of augmentation and in the dose required to produce maximum OFR generation. PMID- 8711265 TI - Effect of luminal calcium on proximal tubule bicarbonate reabsorption in the rat. AB - In previous investigations, it was found that rats depleted in parathyroid hormone (TPTX rats) had reduced rates of proximal bicarbonate reabsorption independent on blood calcium levels. In the present work, the role of calcium (Ca2+) in rat proximal tubule bicarbonate reabsorption was studied by in vivo stationary microperfusion. Tubules were perfused at different lumen Ca2+ concentrations in the presence and absence of the calcium ionophore A23187. Bicarbonate reabsorption was not affected by Ca2+ in the range of 0 to 1 mM, but was significantly reduced when 0.5 mM EGTA was added to the 0 Ca2+ perfusates, indicating that only at very low luminal Ca2+ levels, bicarbonate reabsorption ( = H+ secretion) was impaired. These observations indicate that Ca2+ in the tubule lumen is important for the maintenance of normal proximal bicarbonate transport, but the low Ca2+ level necessary to impair this transport mechanism is achieved only in the presence of EGTA, a condition that simulates the absence of parathyroid hormone. PMID- 8711267 TI - Ultrapure polymerized bovine hemoglobin improves structural and functional integrity of the isolated perfused rat kidney. AB - Since it became evident that organ dysfunctions after acute hemolysis are not induced by hemoglobin per se, but by stroma-contaminated hemoglobin, solutions of ultrapure stroma-free hemoglobins were regarded to be possible substitutes for blood in transfusion medicine. We tested one of the recently developed modified bovine hemoglobins (Ultrapure polymerized bovine hemoglobin 1; UPPBHb1) in the isolated perfused rat kidney (IPRK) model, using a recirculating system. Control kidneys were perfused with a substrate-enriched Ringer solution containing hydroxyethyl starch (HES) to produce isoncotic conditions. In the experimental group HES was substituted in part by UPPBHb1 (34 g/l). For determination of functional parameters, the kidneys were perfused for 180 min. A separate set of kidneys of both groups was perfusion fixed after 80 min of perfusion which is the period of optimal function. Light and electron microscopic analysis revealed major alterations only for the outer medulla of HES kidneys. Only these suffered from a considerable extent of proximal tubular S3 damage, exhibiting condensed tubular epithelia. In the inner stripe of the outer medulla, which is the zone of greatest sensitivity to damage in the isolated perfused kidney, severe hydropic degeneration, cell detachment, and necrotic destruction of the medullary thick ascending limb were seen in the HES-perfused group, too. In the UPPBHb1 group, the medullary thick ascending limb was well preserved, and S3 showed only a minor degree of damage. UPPBHB1 kidneys were further characterized by the occurrence of intracapillary and interstitial precipitates of UPPBHb1 in inner stripe of the outer medulla and inner medulla. The glomerular filtration rate was significantly higher in UPPBHb1-perfused kidneys (870 +/- 80 vs. 630 +/- 55 microliters/min/g kidney weight for HES). Absolute reabsorption of sodium paralleled the behavior of the glomerular filtration rate. The values for renal perfusate flow and urinary flow rate did not differ significantly between both groups. Renal autoregulation was better preserved in UPPBHb1-perfused kidneys (74 +/- 6% of full autoregulatory response) than in HES-perfused controls (42 +/- 4%). Our results suggest that perfusion of isolated rat kidneys with UPPBHb1 improves kidney function and morphology, providing better oxygenation than in control kidneys. UPPBHb1 does not exert additional nephrotoxic effects on the IPRK that will exceed the noxious potential of the method itself. Thus, it must be concluded that UPPBHb1 may be an oxyphoretic blood substitute with nephroprotective characteristics when compared with nonoxyphoretic substitutes. At least, UPPBHb1 seems to be a promising candidate as oxyphoretic additive to perfusates for the IPRK model. PMID- 8711268 TI - Nifedipine reduces postexercise proteinuria in young volunteers. AB - Previous studies have demonstrated that it is possible to prevent postexercise proteinuria with angiotensin-converting enzyme inhibitors. To determine whether calcium antagonists have the same effect, 40 young healthy volunteers underwent maximal aerobic exercise with and without nifedipine 10 mg per os 1 h before the first or second trial. Urinary excretion of albumin (UAE), transferrin (UTE) and alpha 1-microglobulin (UME) were examined before and after each trial. UAE, UTE and UME were significantly increased after exercise. Nifedipine significantly decreased UAE (p = 0.001) and UTE (p = 0.02) after exercise, and slightly decreased the maximal work load and the basal excretion of albumin. UME was unchanged. Therefore, the results of this study demonstrate that nifedipine administration before exercise significantly reduces postexercise proteinuria. PMID- 8711269 TI - Effect of volume expansion on the natriuretic response to ouabain infusion. AB - Evidence exists that an endogenous substance which inhibits (Na+,K+)-ATPase, in a similar manner to the cardiac glycosides, may have important cardiovascular and renal effects. Whilst ouabain or a closely related isomer has been reported to be present in mammalian plasma, renal effects of ouabain occur only at high concentrations. The effect of expansion of blood volume on the renal response to ouabain infusion was examined in conscious sheep. Five sheep with catheters chronically implanted into the renal artery received four treatment combinations in random order: (I) vehicle (0.15 mol l-1 NaCl) infusion; (II) 500 micrograms ouabain infused into the renal artery over 60 min; (III) 500 ml 6% dextran 70 in 0.9% saline infused intravenously, and (IV) the dextran and ouabain treatments together. Treatment with either ouabain or plasma volume expansion produced modest increases in sodium excretion and urine flow. Treatment with ouabain when combined with plasma volume expansion increased sodium excretion from 82 +/- 30 to 880 +/- 203 mumol min-1 and urine flow from 1.9 +/- 1.1 to 7.5 +/- 1.6 ml min 1. This combination of treatments results in a synergistic rather than additive response. This study indicates that under some circumstances the response of the kidney to inhibition of (Na+,K+)-ATPase can be enhanced and, if inhibition can be demonstrated to occur at physiologically relevant concentrations of endogenous digitalis-like factor, would support a possible physiological role for endogenous digitalis-like factor in the regulation of sodium homeostasis. PMID- 8711270 TI - The power of focused tests to detect disease clustering. AB - Statistical tests have been proposed for determining whether incident cases of adverse health effects are 'clustered' together. Several procedures, termed 'focused', specifically analyse disease surveillance data around pre-specified putative sources of environmental hazard. Little has been done to compare the performance of various proposed methods on actual models of clustering. Analytic power functions are derived for three tests of focused clustering. These functions are based on the probabilistic structure of the clustering tests and do not require simulation. The three tests are compared with respect to statistical power on hypothetical data where monotone multiplicative increases in disease risk near a putative hazard define disease clusters of varying intensity. PMID- 8711271 TI - The choice of test for detecting raised disease risk near a point source. AB - This paper considers the problem of testing for excess risk near a point source of risk S, as might be postulated to exist near a nuclear installation, for example. The data are assumed to be in the form of counts in small areas for which expected numbers of cases have also been calculated by reference to national rates. It is shown how to derive the most powerful test against any given alternative hypothesis; such 'linear risk score' tests are further considered, particularly that which scores each case according to the reciprocal of the rank of the distance from S. These tests are compared with two tests due to Stone for general ordered alternatives and the important distinction is drawn between conditional and unconditional versions of the tests. Their behaviour is illustrated by application to data on childhood leukaemia in relation to nuclear installations in Britain. PMID- 8711272 TI - A class of tests for detecting 'general' and 'focused' clustering of rare diseases. AB - This paper proposes a class of tests applicable to the detection of two types of disease clustering 'focused' and 'general' clustering. The former assesses the clustering of observed cases around the fixed point and the latter does not have any prior information on the centre of clustering. The proposed test for 'general' clustering is a generalization of the index for temporal clustering proposed by Tango in that it adjusts for differences in population densities and also in population distributions among categories of the counfounders such as age and sex. Simulation study shows that the proposed 'general' test outperformed the average distance method of Whittemore et al. in most of the cluster models considered. PMID- 8711273 TI - Non-parametric estimation of spatial variation in relative risk. AB - We consider the problem of estimating the spatial variation in relative risks of two diseases, say, over a geographical region. Using an underlying Poisson point process model, we approach the problem as one of density ratio estimation implemented with a non-parametric kernel smoothing method. In order to assess the significance of any local peaks or troughs in the estimated risk surface, we introduce pointwise tolerance contours which can enhance a greyscale image plot of the estimate. We also propose a Monte Carlo test of the null hypothesis of constant risk over the whole region, to avoid possible over-interpretation of the estimated risk surface. We illustrate the capabilities of the methodology with two epidemiological examples. PMID- 8711274 TI - The map comparison problem: tests for the overlap of geographic boundaries. AB - The quality of environmental studies is often compromised by the use of statistics, such as correlation and regression for example, which presuppose a statistical model, linear or otherwise, between two variables. When investigating hypotheses about relationships among geographically distributed variables, an alternative approach is to measure the amount of boundary overlap. Boundaries are geographic zones of rapid change in the intensity of a variable, and are often of scientific interest in their own right. Examples of boundaries include ecotones, genetic hybrid zones, pollution plumes, and the front of the wave of advance of an epidemic. Boundary overlap describes zones where boundaries from two or more variables coincide, and are useful for evaluating epidemiologic hypotheses relating health to environmental exposures. This paper proposes four statistics of boundary overlap, and explores their performance using simulation models and real data describing ozone concentrations and hospital admissions for respiratory conditions. The statistics are found sensitive to different aspects of boundary overlap, and provide an additional diagnostic tool in the analysis of geographically distributed variables. Overlap statistics are expected to come into increasing use as the installed base of geographic information systems increases. PMID- 8711275 TI - Exploratory tools for disease mapping in geographical epidemiology. AB - In the present paper a new shrinkage estimator of relative risk, useful in disease mapping, is compared with the empirical and full Bayes estimators, using death certificate data for lung cancer (males and females) and breast cancer (females) 1982-1988 in the Emilia-Romagna region (Italy). The estimates are obtained averaging the relative risks obtained from recursive partitioning of the set of observed values. The number of partitions is fixed in advance and behaves as a smoothing parameter, the lower the number the higher the degree of shrinkage toward the overall mean of the observed values. PMID- 8711276 TI - Tests for directional space-time interaction in epidemiological data. AB - Spatial incidence of disease is often recorded with time of occurrence as an ordering label. This ordering can be used to provide distance based tests for joint clustering of cases in space and time. Two different tests are proposed: one where a control disease is available, and the other where only standardized rates within census regions are available. PMID- 8711277 TI - The spatial analysis of covariates in a study of environmental epidemiology. AB - Kanawha County, West Virginia, includes a deep river valley with a large population living in close proximity to many chemical manufacturing centres. One study of this area combined an epidemiologic survey of children attending Kanawha County schools with the measurement of the concentrations of 15 volatile organic compounds at each of the 75 elementary schools. Results have been reported by Ware et al. This paper further examines the associations of the concentrations of the ten individual volatile organic compounds related to chemical manufacturing processes with observed lower respiratory symptoms, generalizing the model of spatial variability and measurement error proposed by Donnelly et al. and using semi-variograms to assess the validity of model assumptions. PMID- 8711278 TI - Bayesian estimates of disease maps: how important are priors? AB - In the fully Bayesian (FB) approach to disease mapping the choice of the hyperprior distribution of the dispersion parameter is a key issue. In this context we investigated the sensitivity of the rate ratio estimates to the choice of the hyperprior via a simulation study. We also compared the performance of the FB approach to mapping disease risk to the conventional approach of mapping maximum likelihood (ML) estimates and p-values. The study was modelled on the incidence data of insulin dependent diabetes mellitus (IDDM) as observed in the communes of Sardinia. PMID- 8711279 TI - Bayesian analysis of space-time variation in disease risk. AB - The analysis of variation of risk for a given disease in space and time is a key issue in descriptive epidemiology. When the data are scarce, maximum likelihood estimates of the area-specific risk and of its linear time-trend can be seriously affected by random variation. In this paper, we propose a Bayesian model in which both area-specific intercept and trend are modelled as random effects and correlation between them is allowed for. This model is an extension of that originally proposed for disease mapping. It is illustrated by the analysis of the cumulative prevalence of insulin dependent diabetes mellitus as observed at the military examination of 18-year-old conscripts born in Sardinia during the period 1936-1971. Data concerning the genetic differentiation of the Sardinian population are used to interpret the results. PMID- 8711280 TI - Traffic accident mapping in Bangkok metropolis: a case study. AB - Results from an analysis of traffic accidents from a study of the police records of four police stations in the Bangkok metropolis are presented. The main emphasis in this study was put on the development of a measure for traffic accident density. The traffic flow was estimated at the various study locations by trained experts. By relating the frequency of traffic accidents found in the police records to the independently estimated traffic flow, a measure for traffic accident occurrence, the traffic accident density, was constructed and estimated for the 47 areas in four study locations. The spatial risk structure was estimated and three risk groups were identified. Each area was classified into its associated risk group, and traffic accident maps for the four study locations in the Bangkok metropolis are provided here. The implication for prevention strategies is discussed. PMID- 8711281 TI - Incidence of leukaemia in young people around the La Hague nuclear waste reprocessing plant: a sensitivity analysis. AB - The objectives are to investigate the incidence of childhood leukaemia in young people around the French nuclear waste reprocessing plant (La Hague, Normandy) for the period 1978-1992, and to check the sensitivity of the results to the choice of disease clustering tests and reference rates. The performance of three statistical procedures are compared: a conventional approach; a Poisson maximum test, and an extraction mapping technique. On the whole, the results are in general agreement and demonstrate the apparent existence of a distinct cluster of childhood leukaemia, contiguous and south-east of the La Hague nuclear facility. PMID- 8711282 TI - MCMC methods for putative pollution source problems in environmental epidemiology. AB - This paper demonstrates the use of the Gibbs Sampler and other Markov Chain Monte Carlo (MCMC) methods in two applications in environmental epidemiology. The first example concerns the application of a Metropolis-Hastings/Gibbs sampler to a Cox process with a direction-dependent cluster variance parameter. The second example consists of the estimation of the posterior (spatial) distribution of a putative location. PMID- 8711283 TI - Spatial variation of natural radiation and childhood leukaemia incidence in Great Britain. AB - This paper describes an analysis of the geographical variation of childhood leukaemia incidence in Great Britain over a 15 year period in relation to natural radiation (gamma and radon). Data at the level of the 459 district level local authorities in England, Wales and regional districts in Scotland are analysed in two complementary ways: first, by Poisson regressions with the inclusion of environmental covariates and a smooth spatial structure; secondly, by a hierarchical Bayesian model in which extra-Poisson variability is modelled explicitly in terms of spatial and non-spatial components. From this analysis, we deduce a strong indication that a main part of the variability is accounted for by a local neighbourhood 'clustering' structure. This structure is furthermore relatively stable over the 15 year period for the lymphocytic leukaemias which make up the majority of observed cases. We found no evidence of a positive association of childhood leukaemia incidence with outdoor or indoor gamma radiation levels. There is no consistent evidence of any association with radon levels. Indeed, in the Poisson regressions, a significant positive association was only observed for one 5-year period, a result which is not compatible with a stable environmental effect. Moreover, this positive association became clearly non-significant when over-dispersion relative to the Poisson distribution was taken into account. PMID- 8711284 TI - Multiple comparisons in disease mapping. PMID- 8711285 TI - [What is your diagnosis? Intercurrent left bundle-branch block in atrial fibrillation]. PMID- 8711286 TI - [Angelus Silesius--physician and mystic]. AB - The priest as physician of the soul and the doctor as physician of the body, this ancient model of christian mentality was adapted by Johannes Scheffler-better known under his pseudonym Angelus Silesius-both in his biography and his literary works. Born 1624 in Breslau, he graduated to a Doctor philosophiae et medicinae and became physician in ordinary to an orthodox-Lutheran earl. Contacts to the esoteric Christianity, which was based on theosophy, mysticism and the physics of Johann Bohme, let Scheffler quit his profession. He converted to Catholicism in 1653 and was ordained as a catholic priest in 1661. His two main manuscripts, the Cherubinische Wandersmann and the Heilige Seelenlust, a poetical dyptichon, show the two possibilities of reaching the mystical recognition of god, either the capability of reason or the ability of emotion. The centre of all is the unio mystica, the union with god. Its a mysticism of the spontaneity of salvation, which occurs in a variety of prospects, metaphors and paradoxical language. PMID- 8711287 TI - [Environmental medicine]. AB - As technology and industrialization have altered our environment, we have become increasingly aware that man made environmental change may lead to disease. Obvious examples are the widespread use of asbestos during this century, the epidemic of cigarette-caused disease, and the prevalent air and water pollution. Such examples have lead to more general concerns about the environment, particularly about its global change, and the need for ongoing evaluation of suspect risk factors. Hereby, medicine concerned with these risk factors, environmental medicine and hygiene, involves prevention, diagnosis, therapy, and study of disease and injury due to external influences; its core disciplines are toxicology, epidemiology, and occupational medicine. Yet more important, the environmental medicine is not understood as a new specialty but as a transdisciplinary task of all doctors and those who are concerned with human health problems. PMID- 8711288 TI - [Education in environmental medicine: practical aspects]. AB - Rarely physicians educated in environmental medicine will be able to work as independent specialists. Most probably they will fulfill their mission besides a main job, usually in general medical care. On one hand this additional education may be applied in daily practice (diagnosis and investigation of environmentally induced diseases and disturbances) on the other hand by presentations in schools, courses, public meetings. Of special importance are political activities such as counselling, discussions with authorities of the environmental protection agency and other authorities, official statements etc. aiming at general medical prevention. Different topics of environmental medicine are outlined and the special importance of interdisciplinarity in thinking and research is stressed. Finally some informations for the practitioner concerning postgraduate education and literature-search are given. PMID- 8711289 TI - [Physicians and politics]. AB - In addition to their original healing-mission, physicians gain more and more responsibility in health politics including most important aspects like education and medical ethics. Recent developments and possibilities in prenatal diagnostics, molecular biology and in-vitro-fertilization demand clearcut positions from the medical staff. Political evaluation of costs in health care systems concerns analysis of the most common factors responsible for the increasing costs. High expectations and demands of the patients are one reason, other are excessive use of new technologies, increase of the aged in the population and growing cost for liability cases. Mechanisms for the down regulation of costs are restricted. The so called free market in health care such as premium-competition of health insurance point out more theoretical than practical aspects. The Health insurance law (KVG) by increasing the extent of services has contributed to a further increase of costs instead of lowering them. Shifting from hospitalization to out-patient treatment and reducing beds for acute services could lower hospital costs further. AIDS, Drug-politics and environmental destruction, three actual plagues, represent further political challenges for physicians. For the future a reasonable politic involving a better motivated medical corps has to be promoted. To this end first personal structures should be strengthened by help of new ethical view points. Future medicine should be complex, patient-oriented-and remain affordable. Therefore medical responsibility in political development is mandatory. PMID- 8711291 TI - [Can medicine still be financed? The economist's viewpoint]. AB - Will we be able to finance the healthcare system over the next years? Where are the problems? Do we want to finance a healthcare system, hospitals, physicians or health? There are a lot of causes for the explosion of costs in the public health system. The recommendations of the economists concerning this fact are simple. But the anonymous market or other incentive systems are not sufficient to control the public health system. There are undesirable consequences of these instruments. We will have to accept that allocation policy is responsible for public health. How much satisfaction of what needs will we give up, and who should decide on that subject? At last we will have to learn that life is not for sale and its prolongation is not possible at any price. PMID- 8711290 TI - [Can medicine still be financed? A health director's viewpoint]. AB - The author follows the question how long a medical care system which accommodates new international developments remains affordable. In his opinion this is possible as long as the advantages of modern medicine are valued positively by the public efficiency is proven and expenditures for other needs are no markedly touched. Health care professionals can contribute to the maintainance of financial means by promoting medical progress, by maintaining efficiency and, finally, by fulfilling their role as citizens also in political discussions. PMID- 8711292 TI - [Respiratory allergies: some medical history elements are useful to the practitioner]. PMID- 8711293 TI - [Clinical manifestations of actinomycosis: literature review apropos of a case]. PMID- 8711294 TI - [Pericarditis and other cardiac disorders, late consequences of radiotherapy]. PMID- 8711295 TI - [Chronic cardiac insufficiency: current treatment and future outlook]. PMID- 8711296 TI - [Laryngo-tracheal-bronchial foreign bodies]. PMID- 8711298 TI - [Local anesthetics: indications and complications]. PMID- 8711297 TI - [Adult-onset Still's disease]. PMID- 8711299 TI - [Multiple cholesterol emboli]. PMID- 8711300 TI - [Focus: fulminant and subfulminant hepatic insufficiency]. PMID- 8711301 TI - [[Emergency and Resuscitation Emergency Mobile Service--Neuchatel experience]. PMID- 8711302 TI - [Occupational therapy:neither marginal nor exclusively hospital]. PMID- 8711303 TI - [The treatment of ascites: a medical controversy?]. PMID- 8711304 TI - [Response to "Point de vue" in the Revue Medicale de la Suisse Romande of April 96 (RMSR, 116: 305-307, 1996)]. PMID- 8711305 TI - [Advanced germinal tumors of the testicle: clinical experience with cisplatin, bleomycin and vinblastine (PVB) or etoposide (BEP)]. AB - Thirty previously untreated patients with advanced germ cell testicular tumours received PVB or BEP. Four patients with bulky disease underwent debulking surgery before initiating chemotherapy and two between cycles of treatment. Twenty-seven (90%) complete responses, two (6.7%) partial responses and one (3.3%) no changes were observed. Both partial remissions were rendered disease-free with surgical removal of residual disease. Four patients presented tumour progression with PVB or BEP, three of whom developed a non-germ cell malignancy within the germ cell tumour. One toxic death and three patients with radiological evidence of reversible interstitial pneumonitis attributed to bleomycin were observed. With a median follow-up time of 38.5 months, range 2.5 to 130 months, 83.3% of the patients are alive and free of disease. Actuarial overall survival is 85.6%. This study confirms once again the high percentage of curability of this disease. PMID- 8711306 TI - [Results of the cochlear transplant program of the University of Navarra]. AB - From October 1989 to the end of 1991, 33 patients have received a Cochlear Implant Nucleus 22 Channel, that have been selected from over 150 patients that consulted the possibility of implantation. A cochlear implant program should count with a group of professionals of diverse areas, that work in coordination without losing contact with the rest of the member of the implant team. Cochlear Implant are considered an efficient technique in the treatment of profound neurosensorial deafness, in Pre and Postlinguistic patients. The results obtained have been better in the last group of patients, but results on the whole are considered satisfactory. In prelinguist there is a progressive positive tendency, therefore a longer amount of time is necessary to define the final results. The therapeutic capacity of this method of treatment is not limited only to the auditory area, but it has effect also in other fields related to communication and the integral development of the patients on whom it is applied. PMID- 8711307 TI - [Non-ulcerative dyspepsia with primary functional constipation. Evaluation of its treatment with cinitapride combined or not with methylcellulose]. PMID- 8711308 TI - [Physical exercise and chronic renal insufficiency]. PMID- 8711311 TI - [Palliative medicine before euthanasia]. AB - Paliative Medicine offers many possibilities to reach a good control of the bothering symptoms of terminal patients and to the support of their families. We review the different causes of suffering, suicide and of request of euthanasia as the process of Medical Decision-Making in the care of the Dying. It is necessary to search for ways to reduce the suffering and to improve the quality of life that remains instead of how to finish it earlier. PMID- 8711310 TI - [The new curriculum in medicine]. PMID- 8711309 TI - [5HT3 antagonists: ondansetron]. PMID- 8711312 TI - [Non-invasive diagnosis of rejection in heart transplant: usefulness of Doppler echocardiography]. AB - Cardiac transplantation is a valid therapeutic option for advanced chronic heart failure, with a one year survival of 60-80%. Although the introduction of cyclosporine has markedly improved survival in heart transplant recipients, early detection of cardiac allograft rejection remains the major problem to be solved. At present, the diagnosis of cardiac rejection has been based on the results of endomyocardial biopsy, which remains actually as the gold standard for rejection surveillance. However, ideally the detection of cardiac rejection should be noninvasive to allow frequent follow-up of the patients. Several attempts have been made to find a noninvasive alternative to replace endomyocardial biopsy, doppler-echocardiography being the most promising technique. A review of current noninvasive methods for graft rejection detection will be discussed emphasising the importance of doppler-echocardiography. PMID- 8711313 TI - I International Symposium on Clinical Epidemiology. Sao Paulo, Brazil, May 24-27, 1995. Abstracts. PMID- 8711315 TI - When discharging patients on coumadin, heed this advice. PMID- 8711314 TI - How to find Internet info via snail mail. PMID- 8711316 TI - Ethics in action. Persistent vegetative state. PMID- 8711317 TI - Tradition or science? Spotting the difference in respiratory care. PMID- 8711319 TI - We withdrew nutrition--not care. PMID- 8711318 TI - Restoring electrolyte balance: magnesium. PMID- 8711320 TI - Chemo quick guide. Hormonal agents. PMID- 8711321 TI - Critical care drug update. Part I. PMID- 8711322 TI - An MD's lesson in compassion. PMID- 8711323 TI - Parish nurses help patients--body and soul. PMID- 8711325 TI - Patient-centered care. PMID- 8711324 TI - The nurse's role in police investigations. PMID- 8711326 TI - Minimizing the pain of venipuncture. PMID- 8711327 TI - Nursing with the heart. PMID- 8711328 TI - [A free forearm flap in reconstructive maxillofacial surgery. Surgical technic and results of 40 clinical cases]. AB - The authors report their experience of microsurgical reconstruction of the maxillofacial region with the radial forearm flap in 40 patients. After a brief historical and anatomical description, the cases are presented and specific considerations on the author's operative technique are detailed. The discussion on the advantages and the disadvantages of this flap ends the paper. PMID- 8711330 TI - [Reparative giant cell granuloma. Apropos of a case]. PMID- 8711329 TI - [Mandibular condyle hyperplasia. Therapeutic review]. AB - Excessive cartilage growth on the condyle process of the mandible can result from primary hyperreactivity of the growth cartilage or be a secondary adaptation to an imbalance in occlusive and/or cervicofacial conditions. Treatment depends on the distinction between these two forms. Primary active overgrowth is treated by condylectomy sparing the distal apparatus although conservative surgery to re centering the temporomandibular joint and re-establish symmetry without condylectomy may be used in quiescent moderately active forms saving the joint. For secondary forms, the joint is re-centering and symmetry is re-established without condylectomy. If started early enough, orthopaedic treatment can avoid the development of secondary forms resulting an imbalance in occlusive and/or cervicofacial conditions. PMID- 8711331 TI - [Treatment of a case of multiple tooth agenesis: an orthodontic and implant solution]. AB - Now, the excellent long term results observed in oral implantology allow use of implants in young people. They change habitual prosthetic indications which need healthy teeth mutilations. The authors present a case of multiple dental agenesis in a young girl. She was treated by orthodontic preparation and implants with a good final result (2 years end treatment follow up). PMID- 8711333 TI - [Loss of substance of the lateral region of the face]. AB - The lateral region of the face is often exposed to trauma and may also be the site of tumor localization. Exeresis may lead to major tissue loss. We report several surgical techniques, from the most simple to more complicated, available for facial repair. Neighboring regions in the frontal or temporal area are used but a submental flap may also be useful. PMID- 8711332 TI - [Cherubism: regression of the lesions and spontaneous bone regeneration]. AB - Three cases of cherubism belonging to the same genealogical tree which had been followed up since the age of 7 years to adulthood are presented. These cases prove that, without treatment, the regression of the lesions and bone regeneration are real and not anecdotal. Nevertheless, following spontaneous bone regeneration, radiographically some radiolucent areas of low intensity and devoid of trabeculations, and/or areas of sclerotic bone do persist. In one case, cherubism was associated with peripheral giant cell granulomas localized in areas not affected by cherubism. PMID- 8711334 TI - [Prognostic factors in non-Hodgkin's malignant lymphoma]. AB - A group of 271 patients with non-Hodgkin's malignant lymphomas from years 1975 1989 was tested using multivariant statistical analysis, from the point of view of the prognostic value their basic clinical and laboratory data. In the group of low grade malignancy lymphomas, the following factors showed a prognostic validity: clinical stage, general status, centrocytic histological subtype, and anaemia. In the group of high grade malignant lymphomas following one were set as prognostically important: general status, clinical stage and age. In particular histological subtypes of low grade malignancy statistically valid difference of survival was not proved, the centrocytic type excluded. However, the difference was found between these types and particular subtypes of high malignancy grade. Based on these results, the Cox's risk model was made, enabling us to define 4 risk groups according to the histological subtype and clinical stage: group A (risk < 0.5), group B (risk = 0.5-1.0), group C (risk = 1.0-2.0), group D (risk > 2.0). PMID- 8711335 TI - [Hemostasis disorders in monoclonal gammopathies]. AB - The authors investigated haemostatic parameters of 43 patients with monoclonal gammapathy with principal diagnosis of multiple myeloma (MM-35), non-Hodgkin's lymphoma (NHL-3), M. Waldenstrom (3) and monoclonal gammapathy of undetermined significance (MGUS-2). Primary haemostasis defect was found in 24 patients. With the exception of 2 thrombocytopenic patients, the defect of aggregation, procoagulant activity and retraction is supposed to be caused by paraprotein. The examination of the 14C 5-HT release and TX B2 synthesis in thrombocytes showed, that platelet activation is unaffected and the paraprotein interferes with interactions of thrombocytes or with coagulation system. In plasma coagulation system explicit abnormalities were found only in thrombin time in 12 patients. The more detailed examination disclosed, that the defect resulted from paraprotein interference with fibrin monomer polymerisation. The thrombin proteolytic activity remained unaffected. In 3 patients shortened euglobulin lysis time was observed. Laboratory haemostasis defect was found in 26 patients (60%), however, the bleeding symptoms manifested in 5 cases (11%) only. The analysis of study results showed, that the most important abnormalities leading to overt bleeding are thrombocytopenia or combined haemostasis defect. Isolated laboratory defects remained silent in most cases. PMID- 8711336 TI - [Changes in the pharmacokinetics of trimecaine in patients with liver cirrhosis]. AB - Trimecaine (Mesocain, Leciva) an analogous preparation to lidocaine is used as a local anaesthetic, antiarrhythmic and spasmolytic preparation. The purpose of the investigation was to assess the fate of trimecaine in nine patients with compensated cirrhosis of the liver. The pharmacokinetics were evaluated after intravenous infusion of trimecaine administered at a rate of 150-200 mg.h-1. Plasma concentrations of the drug evaluated by gas chromatography indicate a marked retardation of trimecaine elimination in patients with liver damage. In the group of patients with cirrhosis of the liver the total clearance of the drug was lower (8.7 +/- 5.0 1.h-1), as compared with the group of patients without liver damage (34.7 +/- 19.4 1.h-1). In patients with cirrhosis of the liver also a multiple reduction of distribution volumes was observed. The results indicate a high hepatic extraction of trimecaine. The reduced hepatic flow in patients with cirrhosis of the liver thus calls for a reduction of the booster and maintenance dose. PMID- 8711337 TI - [Treatment of kidney calculi using shock-wave lithotripsy with ultrasonic guidance]. AB - Lithotripsy by means of an extracorporeal shock-wave was performed in 128 patients with urolithiasis. In this group for the first time in Czechoslovakia ultrasound control of kidney stones was used in 44 patients; in the remainder X ray control was used. The authors used equipment designed and manufactured locally. The ultrasonic probe is laterally connected with the shock-wave applicator. Disappearance of the fragments after lithotripsy was achieved in 39 patients where ultrasonic control was used. The paper presents the results, discusses the advantages and limitations of ultrasonic control in extracorporeal lithotripsy of urolithiasis. PMID- 8711338 TI - [Endoscopic sclerotization of esophageal varices and nitrate therapy in portal hypertension associated with esophageal variceal hemorrhage]. AB - The authors investigated changes of some haemodynamic parameters after sclerotization of oesophageal varices and after administration of isosorbide dinitrate in the preparation Iso Mack retard. Twenty patients with cirrhosis of the liver with oesophageal varices grade 3-4 were treated by endoscopic sclerotization only, while to a second comparable group of cirrhotic patients from the first stage of sclerotization oral Iso Mack retard was administered in daily doses of 80-120 mg, depending on tolerance. The haemodynamic parameters were investigated before the onset of treatment and again after completed sclerotherapy or at least three-month treatment with ISDN. After sclerotization of oesophageal varices the pressure in the portal circulation rises significantly as well as the flow through the portal vein due above all to an accelerated blood flow. On the other hand, ISDN prevents a rise of portal hypertension and leads even to a slight decline which is, however, not associated with a decline of the portal flow. A relative disadvantage is the adverse effect on systemic haemodynamics after large nitrate doses. The decline of the portal pressure caused by the vasodilatating effect of ISDN in the splanchnic and portal area is manifested in a very positive way by reduction of relapses of haemorrhage from oesophageal varices. ISDN may play a positive role in the treatment of portal hypertension with oesophageal varices, during acute haemorrhage as well as in the prevention of relapses of haemorrhage. A particularly favourable procedure is the combination with sclerotherapy of oesophageal varices. None of the procedures leads to reduction of the portal flow and preserves hepatic function. PMID- 8711339 TI - [Analysis of the polypeptide profile of insulin preparations using HPLC]. AB - The purity of insulin preparations which is important during clinical administration of this hormone can be assessed in several ways. The authors suggested and tested a method for analysis of the polypeptide composition of insulin preparations which makes quantitative and qualitative evaluation of the insulin and non-insulin character possible, as well as their separation according to the shape and size of biological macromolecules by gel filtration on HPLC equipment. The described procedure makes possible a more detailed analysis of insulin preparations when the method is used in combination with electromigration and immunological estimations. PMID- 8711340 TI - [Distal renal tubular acidosis in patients with Wilson's disease]. AB - Renal affection is part of the clinical picture of developed Wilson's disease. The most frequent sign of affection of the distal nephron is distal tubular acidosis (DRTA), more frequently its latent form with a normal systemic pH, but inability to reduce the urinary pH below 5.5 after an acid load. More detailed assessment of the type of this disorder was not made so far. To elucidate in greater detail the pathogenesis of DRTA in Wilson's disease the authors examined renal acidification in 13 patients with bioptically verified Wilson's disease, using three acidification tests: 1) the test involving oral administration of calcium chloride (CaCl2), 2) the test involving infusion of sodium sulphate (Na2SO4) and 3) the test using an infusion of NaHCO3. In none of the examined patients with Wilson's disease manifest (complete) DRTA was found with systemic acidosis, however, in five patients latent (incomplete) DRTA was revealed, i.e. after administration of CaCl2 the pH of urine did not decline below 5.5. Patients with Wilson's disease and DRTA differed from patients with Wilson's disease and normal renal acidification by the age at the time of assessment of the diagnosis (30.4 +/- 3.6 vs. 20.75 +/- 2.14 years, p < 0.02) and the duration of the disease (i.e. the time from assessment of the diagnosis to the acidification examination: 4.6 +/- 4.86 vs. 15.38 +/- 2.40 years, p < 0.01). In three of four patients with Wilson's disease and DRTA who were given Na2SO4, after its administration the pH of urine did not decline below 5.2. After alkalination of the urine with sodium bicarbonate in all patients with Wilson's disease the pCO2 in urine was higher than 9.1 kPa and the gradient between pCO2 in urine and blood (U-B gradient of pCO2) was higher than 3.32 kPa. Latent (incomplete) DRTA is thus a frequent component of the clinical picture, in particular in Wilson's disease when diagnosed late. The disorder does not respond to administration of Na2SO4, the response of pCO2 to NaHCO3 administration is, however, normal. The results indicate that from the pathogenetic aspect probably the so-called gradient type is involved, characterized by the inability of the acidifying tubular cell in the distal nephron to secrete hydrogen ions against a high pH gradient. PMID- 8711341 TI - [Hypertension in patients with polycystic kidneys--the effect of volume expansion]. AB - Arterial hypertension is found in as many as 75% patients with autosomal dominant polycystic kidneys with normal renal function, its pathogenesis is however not quite clear so far. The authors examined 16 patients with polycystic kidneys with normal or only slightly reduced renal function (plasma creatinine lower than 140 umol/l), 8 of these patients were normotonic (N) and 8 hypertonic (H). In all examined subjects right-sided cardiac catheterization was performed with assessment of the minute cardiac volume by thermodilution. To all patients in the course of one hour 1500 ml saline per 70 kg body weight were administered and the haemodynamic examinations were repeated after termination of the infusion. In all subjects before and after expansion the plasma renin activity was assessed (PRA), as well as plasma aldosterone (PA), plasma catecholamines (PC) and the atrial natriuretic factor (ANF), the renal blood flow and glomerular filtration by means of clearance and extraction of PAH and inulin clearance. The authors did not find differences in plasma concentrations, cardiac output and splanchnic and renal ANF extraction groups N and H, nor in PRA, PA and PC. Volume expansion led in both groups to a comparable rise of ANF and suppression of PRA and PA. Group H did not differ from group N in any of the investigated haemodynamic and renal parameters except for systemic vascular resistance. In hypertensive patients before expansion a close correlation was found between pressure in the pulmonary artery in a wedged position and diuresis (r = 0.935, p < 0.01) and natriuresis (r = 0.895, p < 0.01). The volume expansion was in both groups associated with a comparable rise of diuresis, the haemodynamic response of patients N and H was however quite different. While in patients of group N a decline of the systemic vascular resistance occurred as well as an increase of the minute volume without a change of the renal flow and glomerular filtration, in hypertonic patients the systemic vascular resistance and minute volume did not change but there was a significant rise of the renal flow and glomerular filtration. The relationship of diuresis and natriuresis of hypertensive patients with polycystic kidneys to volume parameters and the rise of the renal perfusion pressure during volume expansion indicates the importance of pressure natriuresis for ensuring the sodium and volume homeostasis in these patients. PMID- 8711342 TI - [Cerebral ischemia attacks from the viewpoint of the clinical neurologist]. AB - A transient ischaemic attack (TIA) is a cerebrovascular attack the clinical symptomatology of which is restored to normal within 24 hours. Minor haemorrhages, epilepsy, hypoglycaemia, migraine and even peripheral neuropathies can be mistaken for TIA. CT examination indicates in a considerable percentage of examined subjects encephalomalacia. The term TIA should be reserved for ischaemic attacks, where after regression of the clinical symptomatology within 24 hours no structural changes are found. PMID- 8711344 TI - [Fenestration of the anterior cerebral artery. Personal experience]. AB - The authors demonstrate a patient with fenestration of the anterior cerebral artery. The fenestration is in the horizontal portion of the anterior cerebral artery (A1). Its embryonic origin and clinical importance arouse discussion. The paper reviews the literature on the subject. PMID- 8711343 TI - [Computer tomography of cerebral ischemia infarct in the diagnosis of transient cerebral ischemia]. AB - The term transient ischaemic attack (TIA) does not correspond to findings of new examination methods and is rather a matter of convention. The authors examined 19 patients with the diagnosis of TIA by CT and found in five a cerebral infarction. The dependence of cerebral infarction in TIA on age, sex, risk factors (hypertension, diabetes) and the duration of clinical symptomatology was not significant. PMID- 8711346 TI - Anticonvulsant action of carbonic anhydrase inhibition. AB - In this paper we review our data from in vivo and in vitro experiments with mutant, carbonic anhydrase II (CA II) deficient mice (Car2n/Car2n mutants) compared to their nonmutant littermates (Car2n/+ or +/+). In vivo, mutant mice were more resistant to flurothyl-, pentylenetetrazol, and loud sound-induced seizures than normal littermates. The increased resistance to flurothyl seizures was age dependent for clonic seizures, occurring after 19 days of age and disappearing after 90 postnatal days. In in vitro experiments, synaptic transmission in hippocampal slices from mutant mice were more resistant to hypoxia than synaptic transmission in slices from normal littermates. There was almost no difference in hippocampal CA1 long-term potentiation of synaptic transmission between mutants and nonmutants. However, studying in vitro epileptogenesis, we found hippocampal slices from mutants to be more prone to seizures in the low Mg2+ environment than slices from normal littermates. This striking difference between in vivo and in vitro seizures susceptibility in CA II deficient mutants suggests and existence of an anticonvulsant factor present in conditions in vivo, but not in vitro. We suggest that extracellular proton concentrations (extracellular pH) acting as N-methyl-D-aspartate (NMDA) receptor antagonist may be such a factor. Mutant mice suffer from severe systemic acidosis that can decrease NMDA receptor function and thus be anticonvulsant in vivo. However in vitro, the steady pH of perfusing solution is relatively alkalinic for mutant mouse slices enhancing the thus NMDA receptor conductance and leading to proconvulsant effects. Thus, the anticonvulsant action of CA inhibition in vivo may be mediated by acidotic extracellular pH rather than an accumulation of CO2 as suggested previously. PMID- 8711345 TI - [The effect of adrenotropic agents on lipolysis in rat adipose tissue; the effect of nonselective alpha adrenomimetics and alpha adrenolytics under in vitro experimental conditions]. AB - It is now generally accepted, that in most of the animal species the adrenergic control of lipolysis from adipose tissue is mediated by adrenergic beta 1 and alpha 2 receptors. Therefore, the problem we deal with in this article is concerned with the role of alphaadrenergic receptors in lipolysis in the rat adipose tissue because only in this species the function of alpha adrenergic receptors in lipolysis is still not clear. In in vitro experiments with epididymal adipose tissue of adult rats (where we followed up the release of free fatty acids into the albumin medium) we found: 1) Alpha adrenergic blocking agents phentolamine and phenoxybenzamine had no influence on the lipid mobilizing effect of adrenergic agonists. This effect was absent also when drugs with the strong alpha sympathomimetic effects, e.g. norepinephrine and noroxedrine, were used. 2) On the other hand, alpha adrenergic agonist phenylephrine was able to antagonize in rat adipose tissue the lipid mobilizing effect of beta adrenergic agonist isoproterenol. 3) From our experiments it can be concluded, that phenylephrine acts as a competitive dualist on beta adrenergic receptors, it reduces the effect of full agonist (e. g. isoproterenol) to the level of its own maximum lipolytic effect. No participation of alpha adrenergic receptors in described effects of phenylephrine were detected, because neither the lipolytic, nor the lipolysis blocking effects of phenylephrine were influenced by alpha adrenergic blocking agents. Our studies using alpha adrenergic agonist phenylephrine or nonselective alpha adrenergic blocking agents did not indicate the presence of any alpha adrenergically controlled lipolysis in the rat adipose tissue. However, the role of antilipolytic alpha 2 adrenoceptors in rat adipose tissue cannot be completely excluded, some of these recaptors were found on the membranes of rat adipocytes. Our future study deals with this problem. PMID- 8711347 TI - Nephrotoxic effects of platinum cytostatics--preventative effects of nifedipine and cimetidine. AB - The study focuses on the observation of nephrotoxic effects of cis-platinum (cis dichloro-diaminoplatinum, CDDP) and carboplatinum (cis-diamino cyclobutancarboxyplatinum, CBDCP) and these to affect them by applying Ca-channel blocker (nifedipine) and a selective antagonist of tubular secretion of organic cations (cimetidine). The urine levels of tubular enzymes alkaline phospatase (AP), gamma-glutamyl-transferase (GMT) and N-acetyl-beta-D-glucosaminidase were selected for the evaluation or tubular toxicity. The clearance of endogenous creatinine was observed as well. The experiments were carried out on male rats of outbred Wistar strain within 7 days after the cytostatics had been applied. A significant increase of urine levels of all enzymes studied was observed in animals, which were given CBDCP only. A maximum change was observed in the urine excretion of NAG. Nifedipine application decreased the change significantly, cimetidine had no effect. The effect of combined nifedipine and cimetidine application did not vary from the effect of application of nifedipine itself. In animals given CDDP only, a significant increase of urine levels of all studied enzymes was proved, the maximum change was observed in NAG excretion. This change significantly decreased by nifedipine application. The same effect was reached by cimetidine application. Preventive effects of nifedipine and cimetidine potentiated mutually each other. In conclusion we consider nifedipine as a convenient prevention of CDDP and CBDCP nephrotoxic effects while cimetidine can be used as a prevention of CDDP nephrotoxic effect only. PMID- 8711348 TI - Triplicity of the malignant tumor of the urinary bladder. AB - Described report case of the patient after left nephrectomy, performed 18 years ago, for adenocarcinoma. Recently he has a simultaneous carcinoma of the bladder and the left ureter. Carcinoma of the bladder was treated with transurethral resection and electrocoagulation. Once checked, the hemorrhage was found from the left ureter, later, papilocarcinoma was diagnosed and the urethrectomy with the resection of the part of the wall of the bladder in the region of the estuary. Patient was in good condition 5 months after the operation. There is a review of others report cases. PMID- 8711349 TI - [The effect of estradiol, norethisterone and testosterone on regional circulation and relative weight of the testes, ovaries and uterus in rats]. AB - In nine experiments of 188 rats (85 males and 103 females), relative weight and local blood flow (with 85Sr-microspheres) was estimated in testes, ovaries and uterus after the administration of estradiol benzoate (Agofollin Depot 1 mg per rat or 5 mg per kg s.c. in time intervals of 3 to 7 days), norethisterone (Norethisterone tablets 0.01 or 0.02% in the food) or testosterone (Agovirin Depot 25 mg per kg s.c. every 5 days); all preparations were administered for four weeks. With one exception (norethisterone in males), the results of two experiments performed in the same manner are presented in every experimental situation. After estradiol, the body weight of rats in both sexes is lowered, relative weight of the uterus increased. The local circulatory indicators i.e. 85Sr-microsphere uptake in % dose per g, blood flow in (ml.min-1).g-1 and organ flow fraction % of cardiac output are decreased in the testes and unchanged on the ovaries. As regards the uterus, in the first experiment the 85Sr-microsphere uptake and the blood flow is significantly lower, in the second experiment these values remain unchanged and flow fraction is increased due to enlargement of the uterus. Norethisterone lowers the body weight and relative weight of the testes; in one experiment it increases relative weight of the uterus. Local circulatory indicators are decreased in the testes and inconstantly in the ovaries, whereas in the uterus the decreases are insignificant. Testosterone does not change the body weight. In one experiment, it increases the cardiac output per 100g of body weight; relative weight of the testes is lowered in one experiment and in the ovaries in both experiments, and markedly increased in the uterus. Local circulatory indicators were decreased in the testes, ovaries and uterus but approximately one half of the differences are significant. Thus, with one exception, all statistically significant local circulatory changes after the administration of the hormonal preparations are decreasing; the suppressive action is evident in the local circulation in sexual organs of the animals of the corresponding sex as well as in animals of the other sex. The suppression of the local circulation in testes and ovaries is probably in connection with endocrine regulations; the decrease of microsphere uptake and blood flow per g in the uterus may participate in insufficient adaptation of the blood supply in the enlargement of the uterus after four weeks' administration of estradiol and testosterone. PMID- 8711350 TI - [Treatment of trochanteric fractures of the femoral bone]. AB - The authors present the evaluation of some most often methods of treatment of trochanteric fractures of femur. They analyze the group of 235 patients in the years 1987 - 1991 treated for fractures by the McLaughlin technique and by Ender nailing. The first method was performed on 121 people and the technique of Ender nailing on 97 people. The results and complications of these methods of osteosynthesis are presented in the paper. The experiences of the authors confirm the advantage of Ender nailing in treating fractures of the oldest injured patients. The Ender nailing method of osteosynthesis is the more advantageous, because the operation takes less time and is easier for patients. After Ender nailing, the possibility of functional rehabilitation and walking on crutches is greater. McLaughlin osteosynthesis of these fractures proved the more advantageous in younger injured patients. PMID- 8711351 TI - [Recurrent varices of the lower extremities and their causes]. AB - Surgery of both primary and secondary varicose veins represents a serious medical and social problem. Middle-age patients are primarily affected. Hence, a great attention to this problem is appropriate and the surgery should be performed with great responsibility. On principle, we are opposed to sclerotization of veins as a primary medical treatment. During the surgery of varicose veins, a great attention should be paid to perfect crossectomy with ligation of all branches and leaving a short stump of vein saphena magna. The most important phase of the surgery, however, is the finding incompetent perforating veins (IP) and their correct treatment, since these are, as follows from our work, the most frequent cause of relapses. Incorrectly treated or left-out IP veins play a key role in the ethology of relapses. These IP veins are very often much broader then the original surface varicose veins. Pre-treatment examination is extremely important. It is necessary to know the condition of deep vein system and to find or identify IP veins. In no circumstances should the relapse veins (RV) surgery be limited to the deletion of varicose veins only. Reexamination of groin should be a rule. The rule of treating the cause and not the consequence should be valid in the case of RV surgery more then anywhere else. PMID- 8711352 TI - [Adult lysosomal and peroxisomal enzyme diseases and glycogenoses. I. Enzyme diseases with predominant symptoms in the fields of internal medicine, dermatology, orthopedics and ophthalmology]. AB - Thanks to ever increasing incidence of late onset genetic metabolic disorders a comprehensive survey is given of the adult lysosomal and peroximal enzymopathies including adult type glycogenoses. For the sake of maximal practical utility the classical enzyme defect scheme was abandoned. The disorders are referred according to the leading symptomatology by which they enter the diagnostic process. In this part the entities are dealt with having symptomatology belonging to the field of internal medicine, dermatology, orthopedy and ophthalmology. PMID- 8711353 TI - [Adult lysosomal and peroxisomal enzyme diseases and glycogenoses. II. Enzyme diseases with predominant psychiatric and neurologic symptoms]. AB - A survey is presented of adult type neurometabolic disorders with dominant psychiatric and/or neurological and myopathic symptomatology. Also mentioned are the manifest clinical symptomatology in heterozygous state and asymptomatic or oligosymptomatic clinical pictures in homozygotes. PMID- 8711354 TI - [Who is concerned with medical science in Europe?]. AB - On the basis of 34th Meeting of the Standing Committee of the European Medical Research Councils held from 2nd to 3rd June 1994 in Interlaken (Switzerland)- besides the list of EMRC members--a proposal was made for solution of this problem in the Czech Republic that has only been observed in this organization. Government of the Czech Republic, Ministry of Education and Ministry of Health have not yet said anything in response to initiatives submitted by the Czech Medical Research Council. PMID- 8711355 TI - [Informatics education in undergraduate study at the 1st Medical School of Charles University]. AB - History of medical informatics at the 1st Medical Faculty goes to 25 years back. Currently curriculum is divided in two parts. In the first year of studies and obligatory course of computer technique is organized including: computer functions, communications with computers, database systems and text editors. In the 4th year of studies a course in clinical informatics is realized including: principles of computer science, statistical software, artificial intelligence, computer in pedagogy, scientific information, classification in medicine, biological signal and picture analysis, hospital information systems, computer in outpatient ward, clinical decision support, computer simulation in clinical medicine, computers in pharmacology, computer in metabolic care, computers in medical devices. In this way medical students are prepared to be able to use technical means in scientific information management. PMID- 8711356 TI - [The European Health Information and Library Association--EAHIL/AEIBS]. AB - The authors give a short history of the association, beginning with its establishment (1986) and a survey of its activities up to the summer of 1994, and its organization. They write about conditions and terms of membership, about aims and objectives of the association: to improve co-operation among European medical libraries, to raise standards of library and information services as well as professionality of librarians and information officers and their continuous education, especially in regard to new technologies. The authors also emphasize the European character and principles of tolerance that enable a connection of medical nursing, veterinary and pharmaceutical libraries/librarians in these specializations in various countries. It is necessary for information users to learn modern library methods, including information skills for the learning process (libraries of medical faculties). No less important is the role of libraries for research and evaluation of research. A survey of Czechoslovak and Czech participation in EAHIL is also given here. The course of the Fourth European Conference of Medical and Health Libraries on Oslo (from 28th June to 2nd July, 1994) is shortly described. Further library conference activities are also mentioned: 7th International Congress of Medical Librarianship in Washington, DC, May 10-12, 1995; Fourth Nordic Conference for Medical Libraries, Copenhagen (Denmark), August 21-24, 1995; EAHIL Symposium on Cooperation of Medical Libraries, Prague (Czech Republic), September 21-22, 1995; Fifth European Conference of Medical and Health Libraries, Coimbra (Portugal), September 18-21, 1996. PMID- 8711357 TI - [Stanislav Mentl (1894-1981)]. PMID- 8711358 TI - [150 years since the birth of Josef Reinsberg, the founder of Czech forensic medicine]. PMID- 8711359 TI - [Purkinje's contribution to the development of the Czech language with emphasis on scientific terminology]. PMID- 8711360 TI - Bilirubin ditaurate--a sensitive compound for examination of photosensitizers. AB - Bilirubin ditaurate (taurobilirubin) proved to be very suitable for estimation of efficiency of different photosensitizers and for determination of the activity of singlet oxygen. The degree of its bleaching was measured at 450 nm, the excitation light was over 600 nm. Halogen lamps were used as light sources. The decoloration of bilirubin ditaurate was related to the intensity of the light source in the presence of the photosensitizer meso-tetra (4-sulfonatophenyl) porphine (TPPS4). From photosensitizers conventionally used in photodynamic therapy (photosan 3, TPPS4, and Zn and Al chelates of phthalocyanine) chloroaluminium phthalocyanine was most effective. From synthetic dyes of nonporphyrin type thiazine dyes were most active. The presumption is expressed that the efficiency of noncoherent light is comparable to the laser radiation when photosensitizers with large absorption band 600-700 nm are used for photodynamic therapy. PMID- 8711361 TI - The effect of calcium channel blockers on cyclosporine A (Cs A) induced nephrotoxicity in rats. AB - The limiting factor in the therapeutical use of cyclosporine A (Cs A) is its nephrotoxicity, which may lead to renal failure. Cs A nephrotoxicity may present itself as an acute decrease in GFR, or as a chronic renal injury. Nephrotoxicity is caused by the indirect vasoconstriction effect mainly on proximal tubule and afferent arteriols. In our study we have concentrated on the effect of Ca-channel blockers on Cs A nephrotoxicity. As parameters of toxic kidney damage we have used the urine levels of the following enzymes: N-acetyl-beta-D-glucosaminidase (NAG), gama-glutamyltransferase (GMT) and alkaline phosphatase (ALP). Daily intragastric application of verapamil (V) (dose 1.0 mg/kg BW) or nifedipine (N) (dose 0.1 mg/kg BW) was started in a group of male Wistar rats. Cs A (Sandimun Sandoz, Switzerland) was applied daily intraperitoneally 30 minutes after the application of V or N. The dose of Cs A ranged from 5 mg/kg BW to 25 mg/kg BW in individual groups. The animals were observed for 10 days after the drugs application. Urine samples were collected and examined at the end of the whole experiment. The individual parameters were evaluated in the groups receiving the 3 different doses of Cs A (5-25 mg/kg BW). The serum creatinine rose moderately during the experiment. When the Ca-channel blockers were administered, the rise was not as steep, but when the highest dose of Cs A was administered, the Ca channel blockers did not influence the elevation of the serum creatinine. Using the standard dose of Cs A (5 mg/kg BW) the protective effect of Ca-channel blockers can be found. In higher doses of Cs A this protective effect was not expressed. PMID- 8711362 TI - Preventive wire ligature of venae dorsales penis in radical retropubic surgery. AB - The method of preventive hemostasis as suggested by authors is capable of safely arresting the blood leakage into the operation field from spatium Retzii especially in radical retropubic surgery. Retropubic radical surgery requires haemostasis of the dorsal penile veins. In extracapsular prostatectomy, adequate haemostasis permits a safe well controllable urethroyesical anastomosis, diminishing blood loss, shortening the operation time and reducing the risk of postoperative stricture and incontinence development. In cystoprostatectomy, preventive haemostasis reduces the blood loss due to protracted haemorrhage taking place during the neovesica construction and shortens the operation period. In looking for a method of effective preventive ligature of the venae dorsales penis we made use of the technique as described by Berger and Ireton (1). In our opinion, that method is rather complicated requiring and presents requirements for a special instrumentarium and therefore, we regard it as impracticable. Based on the principles of the above technique we elaborated a method of preventive haemostasis using a wire loop to compress both these superficial and the subfascial dorsal penile veins (2), which appears easy to perform with the ordinary equipment of any surgical and urological department. PMID- 8711363 TI - [Surgical complications of vascular procedures in hemodialysis therapy]. AB - The paper presents surgical complications of vascular access in patients with chronic haemodialysis. The author demonstrates his experience with 303 patients having this special vascular operation. The incidence, management, and possible causes of respective complications are reviewed. The most common complications were late thrombosis, infection and aneurysms. The least complications occurred in autogenous arteriovenous fistulas according to Brescia-Cimino. There was no steal syndrome in the whole group of patients. Except an accurate operating technique it is of great importance to take a careful care for and to manipulate exactly with the fistulas in the haemodialysis departments. PMID- 8711364 TI - [The effect of lovastatin on the development of adriamycin nephropathy in rats]. AB - BACKGROUNDS: Hyperlipidemia is not only a prominent complication of nephrotic syndrome, but it may also contribute to the further non-immunologic damage of glomeruli. Analogy between atherosclerosis and glomerular sclerosis was suggested. Treatment of hyperlipidemia may decrease proteinuria in nephrotic animals and subjects and possibly prevent the progression of glomerulosclerosis and renal failure. We decided to study the influence of therapy with the inhibitor of cholesterol synthesis, lovastatin, on the development of the experimental nephrotic syndrome induced by the administration of adriamycine in rats. METHODS AND RESULTS: Nephrotic syndrome was induced in rats by the intravenous administration of adriamycine. One group of animals was treated from the time of adriamycine adminstration with lovastatin. Lovastatin was also given to one group of control animals. Proteinuria increased significantly during 4 weeks in untreated adriamycine rats (from 0.93 + 0.57 to 12.76 + 11.95 g/mmol of urinary creatinine, p < 0.01), but it did not change significantly in adriamycine rats treated with lovastatin (from 0.98 + 0.69 to 1.90 + 4.25 g/mmol of creatinine, p = n.s.) and it was significantly lower than in untreated animals (p < 0.01). Plasma albumin decreased 12 weeks after adriamycine administration in untreated rats (from 21.16 + 1.45 to 10.58 + 3.83 g/l, p < 0.001) significantly more (p < 0.05) than in rats treated with lovastatin (from 20.01 + 2.18 to 15.34 + 2.66 g/l, p < 0.01). Lovastatin also ameliorated the increase of plasma cholesterol and eliminated the increase of plasma triglycerides in adriamycine rats. Metabolic changes were in all groups characterized by the increase of free fatty acids, possibly due to exaggerated lipolysis, but without significant change of glycaemia and plasma urea. Proteinuria was decreasing at the end of the observed period. Histologically there were only minimal changes in glomeruli, without significant glomerulosclerosis. CONCLUSIONS: Administration of lovastatin prevented the development of nephrotic syndrome in experimental adriamycine nephropathy in rats. This finding suggests the possible role of hyperlipidemia in the pathogenesis of glomerular damage and suggests the posibility to prevent glomerulosclerosis and renalisufficiency in some form on nephrotic syndrome by the effective hypolipidemic therapy. PMID- 8711365 TI - [The effect of polyene phosphatidylcholine administration on lipid metabolism and glucose tolerance in patients with hyperlipoproteinemia IIB]. AB - BACKGROUND: Up to now, clinical and experimental studies have brought only inconsistent data about the effectiveness of polyene phosphatidylcholine (PPC) in treatment of dyslipidemia. The aim of this randomized, double blind study was to verify the effects of the polyene phosphatidylcholine preparation Lipostabil forte R (Rhone-Poulenc-Rohrer, Koln, FRG), given orally in a daily dose 2.7 g for three months to patients with hyperlipoproteinemia type IIB and hypoalfacholesterolemia both on the plasma lipids, lipoproteins, apolipoproteins and the parameters of glucose tolerance. METHODS AND RESULTS: Administration of PPC to 30 patients with hyperlipoproteinemia type IIB and hypoalfacholesterolemia led to a significant rise of HDL-cholesterol, HDL3-cholesterol and plasma apolipoprotein A-I concentrations in comparison with the group treated with placebo. At the same time, plasma apolipoprotein B concentration slightly increased. Blood glucose, immunoreactive insulin and non-esterified fatty acid concentrations during the oral glucose tolerance test didn't change significantly after PPC administration. CONCLUSIONS: It seems PPC could be the appropriate supplement to the treatment of patients with decreased concentrations of HDL cholesterol and plasma apolipoprotein A-I. PMID- 8711367 TI - [Amusia]. PMID- 8711366 TI - [5 years' experience with conscious sedation with midazolam (Dormicum Roche) in stomatology]. AB - The authors used Midazolam in dental treatment of 225 patients of both sexes (ASA I-IV). 25 patients suffering from heart diseases (ASA III-IV) were ECG monitored during the treatment. Midazolam with the biological half-life of 0.5-1.5 hours was successfully used for premedication of dental out-patients. The used doses had no significant influence on hemodynamic and respiratory values. Specific antagonist Flumazenil reduced the period of psychological recovery of the patients and thus made it possible for the patients to be dismissed within 30 minutes after the dental treatment if accompanied. PMID- 8711368 TI - [I just graduated and what i think about it]. PMID- 8711369 TI - [Professor Jaroslav Jedlicka, Dr.Sc. (1891-1974)]. PMID- 8711370 TI - Meso-tetra-(4-sulfonatophenyl)-porphine of low neurotoxicity. AB - The acute toxicity of specially purified meso-tetra-(4-sulfonatophenyl)-porphine (TPPS4) was measured in mice DBA/2 and Wistar rats after i.v. administration of the dye dissolved in saline solution. LD50 of our preparation was 352.5 mg/kg for mice and 574.9 mg/kg for rats. Motor nerve conduction velocity was measured in rats and rabbits in ether anesthesia N. ischiadicus and n. tibialis were stimulated by surface electrodes and M response was registered. No signs of neurotoxicity were found, even after application of 150 mg/kg b.w. of our TPPS4. Histologic evaluation was performed by routine method and standard electron microscopic procedure. Only minimal and sporadic axonal changes were found. The clinical application of low toxic TPPS4 was reconsidered. PMID- 8711371 TI - Anabolic effects of progestogens in bone remodelling. AB - The short-term effects of oestrogen deficiency on biochemical indices of bone turnover were studied in 49 women who had undergone hysterectomy with bilateral ovariectomy. In patients treated with norethisterone or tibolone, a significant reduction was observed in urinary hydroxyproline excretion comparable to that observed in estradiol treated patients. Bone isoenzyme of serum alkaline phosphatase activity did not decrease and bone mass remained increased during norethisterone and tibolone treatments. It is concluded that norethisterone and tibolone can act as bone-trophic hormones. PMID- 8711372 TI - Somatometric and hormonal effects of dexfenfluramine. AB - The effect of dexfenfluramine, the new serotoninergic appetite suppressor, was tested in 20 women. The drug was administered 3 months, 15 mg twice a day. A selected reduction of carbohydrate and of fat intake was observed. The body weight decreased mainly due to fat loss in the abdominal region. An important decrease of blood lipids was recorded, while insulinemia and OGTT remained unchanged. Serum alkaline phosphatase increased and ALT decreased. Other liver tests did not change. During the treatment the level of cortisol, TSH, T3 and 17 ketosteroids were not affected. T4 decreased and GH increased. These metabolic and hormonal effects were discussed. The treatment with dexfenfluramine was well tolerated and was found to have several favourable effects. PMID- 8711373 TI - Dehydroepiandrosterone and insulinaemia. AB - Low dehydroepiandrosterone (DHEA) and hyperinsulinaemia are assumed to be risk factors of atherosclerosis. Exogenous hyperinsulinaemia during hyperinsulinaemic clamp decreases DHEA. We have evaluated interaction of these hormones during 2 weeks of therapeutic starvation in 11 obese patients (mean BMI 41.2 kg/m2), 5 nondiabetics and 6 diabetics with diet only therapy. Comparing diabetics with normals we have found no differences in BMI and blood DHEA, DHEAS and insulin levels. We have found a light implication of negative correlation of insulin to DHEA and DHEAS level in diabetics (r = -0.71, -0.73, p = 0.16, 0.18). During starvation insulinaemia is declining, DHEA initial increase is followed by a decrease (r = 0.93, p = 0.01). We conclude that insulin decrease during starvation could be a cause of DHEA decrease. This finding is unexpected according to the potential of exogenous insulin to suppress DHEA. Further investigation of endogenous DHEA and insulin relation is necessary. PMID- 8711374 TI - Retrospective testing of an insulin advisory system. AB - A retrospective analysis of the function of adaptive model based consultation system for insulin therapy was performed. The program proved good adaptation ability with improving blood glucose level prediction. In a group of 12 brittle type 1 diabetic patients individual analysis showed problems resulting from retrospective testing of dynamic systems, when treatment is determined only by the physician. PMID- 8711375 TI - [A decrease in blood flow and incorporation of 45Ca and 3H-proline in the bones in pregnant female rats]. AB - In the experiments on rats, the bone blood flow may be influenced by the deficiency as well by the administration of sex hormones. The aim of this work was to investigate the bone circulation (determined by means of 85Sr microspheres, experiment A and B) and the incorporation of 45Ca and 3H-proline into the tibia (experiment C) in the last stage of pregnancy (from 18 to 21 days), i.e. in the physiological condition with elevated level of estrogens. In the experiment A and B, the uptake of 85Sr-microspheres and blood flow in pregnant rats was lower in the tibia and distal femur, the blood flow only was significantly lower also in the humerus and the calvaria. In kidneys, liver, muscle and skin, there was no difference in microsphere uptake; the blood flow was in pregnant females lower only in the kidneys. In perirenal fat, the microsphere uptake and blood flow was several times higher in pregnant rats than in controls. 24 hour incorporation of 45Ca and 3H-proline was in pregnant females significantly lower. The bone density and ash weight was higher in pregnant rats in experiment A. Estradiol concentration in plasma was in experiment B in pregnant rats significantly higher (by 24.8%). Thus, in pregnant female rats, we found lower bone blood flow and lower incorporation of 45Ca and 3H-proline along with normal or higher bone mineral content in the condition, where enhanced resorption and formation of the bone had been described. The changes in bone blood flow and possible role of estrogens in this situation remain to be elucidated. PMID- 8711377 TI - Multicomponent analysis of biological fluids with the arrays of sparingly selective ion-selective electrodes. PMID- 8711376 TI - Methylene blue--an endocrine modulator. AB - Methylene blue (MB) is a thiazine dye used in the treatment of methemoglobinemia. It may represent a new class of anti-oxidant drugs which competitively inhibit the reduction of molecular oxygen to superoxide by acting as an alternative electron acceptor for tissue oxidases. Because of its strong free radicals scavenging effect MB was experimentally used in the treatment of reperfusion syndrome. MB is soluble guanylate cyclase inhibitor. It was found to inhibit the stimulation of soluble guanylate cyclase by nitric oxide and vasodilatators. Another effect of MB is inhibition of prostacyclin synthesis by endothelial cells and isolated arteries independently of its effects on cGMP accumulation. We investigated the MB in series of experimental endocrine situations in which its free radicals scavenging effect could play a role. We observed that MB partly inhibited the increase in adenohypophyseal weight, cAMP and blood prolactin levels in male rats after the administration of estrogens. MB also blocked the increase of another free radicals scavenger-the metalloenzyme ceruloplasmin in the blood of estrogenized rats and prevented the increase of bone mineral after estradiol treatment. MB produced a decrease in adenohypophyseal ascorbic acid content. The blood thyroxine levels increased and the anterior pituitary TSH concentration decreased after MB treatment. PMID- 8711378 TI - [50 years since the founding of the 3rd Internal Medicine Clinic of the 1st Medical School of Charles University]. PMID- 8711379 TI - [Clinical neuroendocrinology]. AB - An overview of research in clinical neuroendocrinology at the 3 Dept. of Internal Medicine, Charles University in Prague is reported. The research was focused on diagnosis and therapy of pituitary adenomas and on pathophysiology of some pituitary hormones. The follow up of changes in patients with acromegaly may be used as a model for possible side-effects of growth hormone therapy or doping. In acromegaly not only growth hormone serum levels but also IGF and its binding protein 3(IGF BP-3) is used when evaluating the activity of the disease. In therapy first experience was obtained with radiosurgery using Leksell gamma knife. Thirteen year experience in using a dopaminergic agonist terguride (of Czech origin) in treatment of prolactinomas was completed. Terguride represents the treatment of first choice in patients with prolactinomas of each size, giant prolactinomas inclusively. In patients with central Cushing's disease microsurgical selective removal of pituitary adenoma is preferred. When not possible, the treatment with Leksell gamma knife is indicated with the condition of normalization of cortisol secretion with medicamentous therapy until the full effects of radiosurgery are present. Growth hormone and prolactin were proved to be immunologicaly active hormones, the effect of growth hormone being mediated by somatomedins. Somatomedins seem to be ubiquitous growth factors. Their presence was demonstrated even in eggs. PMID- 8711380 TI - [Experimental neuroendocrinology]. AB - A short review of the history of experimental neuroendocrinology is given. The influence of light on the endocrine system, hypothalamic regulation of the adenohypophysis and neurosecretion are discussed. At the IIIrd Medical Dept. under influence of professor Charvat-I first studied the influence of light (following the findings of Rowan, Scharrer, Benoit and others). After the Harris' discovery that the hypophysial portal system serves for transport of hypothalamic hypophysiotropic factors I relatively early (1949) started to study the hypothalamic extracts, aiming to demonstrate the existence of these factors, the regulating hormones (RH) of today. Our successful attempt to demonstrate TRH (1961) as well the unsuccessful attempt do achieve its chemical constitution are described. Thereafter we studied the mechanisms of adenohypophyseal growth: estrogen-induced adenohypophyseal hypertrophy is inhibited by testosterone, thyroxine and dopaminergic agonists, it is potentiated by dopaminergic antagonists. An inhibitory action is displayed also by methylene blue. There is a correlation between the blood ceruloplasmin (polyphenoloxidase) level and growth of the adenohypophysis after estrogens: methylene blue inhibits both reactions. Finally, we codiscovered the endogenous digitalis-like activity and first demonstrated its origin in the adrenals; we also obtained the preliminary evidence for the existence of quinidine-like endogenous factor. PMID- 8711382 TI - [Obesity and associated metabolic disorders]. PMID- 8711381 TI - Clinical research of glucose metabolism. AB - Glucose metabolism in diabetes and in hypoglycemic states has been studied at IIIrd Medical Department for more than 30 years. The research is now concentrated on the evaluation of insulin action on receptor and postreceptor levels and in biochemical changes accompanying early stages of diabetic microangiopathy. The changes of insulin action has been examined by using the clamp techniques also in patients with organic hyperinsulinism. We study molecular changes of insulin action and pathogenesis of vascular complications. PMID- 8711384 TI - [Osteoporosis]. PMID- 8711383 TI - [Primary hyperparathyroidism]. AB - Over the past 40 years primary hyperparathyroidism (PHP) has changed from a rare, severe disease of the bones and kidneys to a common disease with hypertension, peptic ulcer, pancreatitis, easy fatigue and proximal muscle weakness. We have during these 40 years examined one of the greatest group of patients with PHP. PHP had its maximum incidence in women over the age of 40. The disease is four times frequent in women as in man. The incidence of hypertension and peptic ulcer between patients with PHP is higher as compared with the incidence of these diseases in general populations. The severity of bone changes in individual patients with PHP does not result from the direct action of a single hormone only. Parathyroid hormone (PTH) have hypotensive and vasodilator effects on various vascular beds. The resting blood flow in the limbs of our patients with PHP is increased in comparison with control subjects. PTH increases plasma renin activity in normotensive controls. This effect is partly blocked by beta adrenergic blockers. PMID- 8711385 TI - [Thyropathies]. AB - This text presents a very brief historical overview of the globally significant discoveries in thyroidology. Three areas of thyroidological research are mentioned: questions of autoimmunity, iodine deficiency and the tumorigenesis of the thyroid glands. The following text refers to the colleagues and pupils of Professor Charvat who concentrated on thyroid gland research at the Third Medical Clinic. The contribution of the radioisotopic laboratory which-as part of the hospital-facilitated not only prompt diagnosis of the thyroid diseases of both in and out-patients, but also provided the basis for extensive cooperation and research between specialists at the clinic, i.e. between obesitologists, cardiologists, haematologists, osteologists and paediatricians. PMID- 8711386 TI - [Familial hyperlipoproteinemia and dyslipoproteinemia]. PMID- 8711387 TI - [Clinical cytogenetics]. PMID- 8711388 TI - [Educational innovations at the 3rd Internal Medicine Clinic]. AB - Professor Joseph Charvat, head of the IIIrd Medical Department in 1945-1970, was interested in medical education very broadly and initiated research work in this field. Later in the seventies and eighties his disciples realized a number of educational programmes concerning improving the effectiveness of clinical teaching by the use the method of self-directed learning, assessing students' clinical competence, students' undergraduate education for primary care and computer-aided analysis of undergraduate education results in internal medicine. Recently attention was focused on problems of ethics and physician-patient and health team communication and in this respect a new educational programme of students' early clinical contact in internal medicine for students of preclinical years was established. PMID- 8711389 TI - [Computers in medicine]. PMID- 8711390 TI - [Radionuclides in endocrinology diagnosis and experimental research]. AB - In the article, a brief historical survey is presented of the utilization of radioactive isotopes in endocrinology and, in particular, in the 3rd Medical Clinic and Laboratory for endocrinology and metabolism. In the following, the results of our own work on the local circulation of blood in rats, its regulations and changes, are presented, i.e.: blood flow through the thyroid gland, adrenals and pituitary, local and general circulatory effects of thyroid hormones and, particularly, local blood flow through the bones in rats-changes after castration and administration of sex hormones in both females and males, possible mechanisms of the changes and functional interconnections with regard to the possible participation of the flow changes in the pathogenesis of osteopathies. PMID- 8711391 TI - [Clinical medicine and nursing. Therapy or care?]. PMID- 8711392 TI - [Endocrinology of arterial hypertension]. PMID- 8711393 TI - Three autacoids--endogenous digitalis-like factor, clonidine-displacing substance, and quinidine-like immunoreactivity. AB - The existence of endogenous ligands of opioid receptors prompted research of a potential endogenous digitalis-like factor (DLF) and of endogenous clonidine displacing substance (CDS). Within eleven years of research, endogenous ouabaine was identified as DLF. It originates in the adrenal cortex. Its physiological role is not yet clear. Most probably, endogenous ouabaine is primarily active in regulation of natriuresis and of blood pressure. The CDS originates in the brain and is active in regulation of blood pressure as well. Its chemical formula was recently identified as agmatine. The potential presence of further autacoids like, e.g. endogenous quinidine-like substance remains to be clarified. PMID- 8711394 TI - [Molecular endocrinology of growth factors]. PMID- 8711396 TI - [Geriatrics]. PMID- 8711395 TI - [Methods of saturation analysis in endocrinology]. AB - The article is a brief review of the development of the method of saturation analysis (MSA) from its beginning to nowdays. Our clinic participated at some stages on this development. Sramkova introduced the RIA of GH in 1968, one year later RIA for prolactin and in 1973 RIA for insulin. The extention of the RIA principle to MSA was after its foundation applicated by Justova et al. for the determination of metabolites of vitamin D. Calcitriol, hormon of vitamin D metabolites was analysed by radioreceptorassay (RRA). The same principle of RRA was applicated at the determination of receptors for insulin by Hilgertova. The application of MSA at our clinic is summarized in table 1. and 2. The home made methods and the choice of commercial kits were verified not only from the point of view of the demands on the quantitative parameters of the method but also from the point of view of the cost of a kit. At our clinic we follow the development of MSA of specific binding proteins (BP), trying to introduce or modify the described techniques f. e. the analyses of BP of GH (GHBP), BP for IGF-I (IGFBP 3, IGFBP-1). The extention of the analysed compounds leads to the precision in the diagnosis and more effective monitoring in the process of treatment. Generally, the article describes the utilization of MSA in laboratories interested not only in the routine work, but in the research activity as well. PMID- 8711397 TI - Heart disease in acromegaly. AB - Evidence has accumulated regarding acromegalic heart muscle disease which existence now appears to be unequivocal. We took an advantage of a large group of acromegalic patients being followed-up at our institution for a long time and have studied pattern, nature and reversibility of acromegalic heart disease. Its major manifestation is cardiac hypertrophy expressed especially as left ventricular hypertrophy. The cardiac hypertrophy is slowly reversible after normalization of plasma growth hormone levels due to successful treatment. This we have first suggested on the basis of a retrospective analysis of clinical and echocardiographic data in 78 patients with acromegaly and subsequently confirmed by a 10-year prospective follow-up of the original patient cohort. We have also showed that effective treatment of acromegaly with a new slow release somatostatine analogue lanreotide leads to regression of cardiac hypertrophy. PMID- 8711398 TI - [Endothelin and arterial hypertension]. PMID- 8711399 TI - [New findings in nutritional pharmacology]. PMID- 8711400 TI - [Histomorphometry of bone in clinical practice and research]. AB - Short overview of importance and principles of histology and histomorphometry of bone tissue patterns performed in IIIrd internal clinic since 1978 is described. Since the beginning, 736 bone biopsy patterns were taken. Generally there are two groups of indications for bone tissue investigation: differential diagnosis of osteologic or hematologic diseases, or investigational indication in a research study. Short overview of morphometric results, gained in IIIrd internal clinic is stated. Histomorphometric parameters of acromegalic and normal trabecular bone were compared and results and consequences for architecture of trabecular bone are described. No correlation between levels of somatotropin or somatomedin C and histomorphometric parameters was found out. Principles of further investigation in IIIrd internal clinic are outlined. PMID- 8711402 TI - [Echocardiography in mitral valve valvuloplasty]. AB - Two-dimensional and Doppler echocardiography have an important role in the management of patients with mitral balloon valvuloplasty (MBV). It is extremely useful in the selection of patients for the procedure, the prediction of its outcome, the assessment of the effect of MBV, the recognition of major complications and long-term follow-up of patients with the aim to recognize early restenosis. The authors report their own experience with long-term follow-up of the population of 55 patients after MBV. PMID- 8711401 TI - [Acute phase proteins]. AB - Various plasma protein patterns following inflammation and metabolic stress were described since diagnostic value of ESR was established. This reactive dysproteinemia is now recognized to reflect the shift in hepatic proteosynthesis after the immunoendocrinological activation. Four main groups of mediators are necessary for expression of hepatic positive and negative acute phase proteins (APPs): first and second "wave' of proinflammatory cytokines, further glucocorticoids and growth factors including insulin. Actual data showing details of the immunological and endocrinological regulation of stress hepatocyte proteosynthesis are summarized and our own results are presented. We evaluated the diagnostic use of APPs in some defined clinical situations (neutropenic period after bone marrow transplantation, postoperative complications) and correlated APPs with the plasma levels of the circulating interleukin-6 and cortisol. In another study, APPs, plasma and urinary TNF, interleukin-6, interleukin-8 and adhesive molecules ICAM-1, VCAM-1 were found to be significantly different in various glomerulopathies. Finally, our experimental data indicate the nitric oxide participation in oestradiol regulation of coeruloplasmin synthesis in rat. PMID- 8711404 TI - [Reminiscence about the beginnings of the 3rd Internal Medicine Clinic]. PMID- 8711403 TI - [Vitamin D in clinical practice]. AB - The author summarizes influence of vitamin D and his metabolites in clinical medicine. He draws attention on three eras of vitamin D and future significance of his metabolites and analogues. In this paper are concerned only some of own experiences in this problem performed on Third Medical Clinic 1st Medical Faculty Charles University in Prague, where laboratory measurements of plasma concentrations of metabolites were in Czech republic operating for first time. Vitamin D has a great future due to his extensive effects. PMID- 8711405 TI - [Reminiscence about the beginnings of the 3rd Internal Medicine Clinic]. PMID- 8711406 TI - [50 years' of the 2nd Department of Clinical Chemistry at Charles University in Prague]. PMID- 8711407 TI - Our experience in brachytherapy of certain non-cancerous diseases. AB - Brachytherapy of non-cancerous diseases has been used from the beginning of our century. The analgetic and antiinflammatory effects of gamma rays of the radionuclide Ra-226 has been used in the health resort in Jachymov, located in Ore Mountains, Northern Bohemia since about 1911 in the therapy of patients hospitalized here with rheumatic diseases of joints, tendons and muscles, including the therapy of radical syndromes. The conditions for the therapy by this method are the age of above 42 years, normal blood count and certain other parameters commonly used in the non-cancer brachytherapy. The radiation dose depends on the degree of the involvement. The local single dose in our patients was of 1.8 to 3.5 Gy with an exposure time of 6 hours. In our work, we considered the degree of the damage to the skin on the irradiation site, after single dose, immediately after the application, as well as after the repeated irradiation in the course of several years. In persons followed, we also provided the examination of the blood count before and after irradiation. The results obtained demonstrated that the brachytherapy used in the health resort Jachymov is safe and that it exerts no adverse effects on the patients submitted to it. PMID- 8711408 TI - [Photodynamic therapy of malignant tumors]. PMID- 8711409 TI - High performance liquid chromatography enables fast determination of ratios between individual proteolytic enzymes in human gastric mucosa. AB - The human gastric mucosa contains five isozymogens of pepsinogen (pepsinogens PGA1-PGA5), two isozymogens of gastricsinogen (gastricsinogens PGC6-PGC7) and zymogens of cathepsins. Ratios between some individual pepsins or pepsinogens are very important from a diagnostic point of view. The ratio of pepsinogen 3 to pepsinogen 5 is significant marker of gastric cancer and gastric ulcer. High performance ion-exchange chromatography is an easy and fast method for determination of ratios between individual proteolytic zymogens in human gastric mucosa and thus could serve for additional diagnosis of gastric diseases. PMID- 8711410 TI - [Lipid metabolism in the nephrotic syndrome]. AB - BACKGROUNDS: Hyperlipidemia is a common and serious complication in patients with chronic nephrotic syndrome which unremitted may result in accelerated atherosclerosis. Accumulation of lipids in glomeruli may moreover contribute to the progression of renal insufficiency in these patients. METHODS: Total cholesterol, HDL-cholesterol and its HDL2 a HDL3 subfractions, LDL-cholesterol, triglycerides, free fatty acids, apoprotein B, apoprotein A1 and lipoprotein (a) were determined before any immunosuppressive and/or hypolipidemic therapy in 15 patients (pts) with nephrotic syndrome (8 pts with proteinuria higher than 10 g/24 hrs-NS-A, 7 with proteinuria between 5 and 10 g/24 hrs-NS-B), 7 pts with glomerulonephritis with low proteinuria (GN) and 6 pts with autosomal dominant polycystic kidney disease (PKD). All pts had normal glomerular filtration rate. RESULTS: Plasma cholesterol (10.26 +/- 2.22 vs. 4.75 +/- 0.66 mmol/1, p < 0.001), plasma triglycerides (4.03 +/- 1.64 vs. 2.17 +/- 0.12 mmol/1, p < 0.05), LDL cholesterol (7.16 +/- 1.55 vs. 2.62 +/- 0.67 mmol/1, p < 0.001) and apoprotein B (1.92 +/- 0.47 vs. 1.11 +/- 0.31 g/1, p < 0.01) were significantly higher in NS-A than in PKD. There was no difference in total HDL-cholesterol and the ratio HDL2/HDL3 between investigated groups of pts. Lipoprotein (a) was higher in NS-A than in other investigated groups of pts. The differences between NS-B and PKD were less significant. CONCLUSIONS: Disorder of lipid metabolism is more serious in patients with more severe nephrotic syndrome than in nephrotic patients with milder proteinuria. Hypercholesterolemia is complicated in these subjects by hypertriglyceridemia and by the increase of apoprotein B and A1 and lipoprotein (a). Metabolic and renal consequences of these disorders depend undoubtedly on their duration and probably also on other factors (e.g. age, hypertension, long term therapy with corticosteroids., etc.). PMID- 8711411 TI - [Further evidence of the role of prostaglandin on the effects of hormones on blood circulation in bones]. AB - In a short communication we demonstrated the inhibitory affect of acetylosalicylic acid (ASA) on the increased bone blood flow and 45Ca and 3H proline incorporation in oophorectomized (OOX) female rats. This finding suggested probable participation of prostaglandin. The aim of the present work was to confirm and extend the initial observation and to find out, whether the administration of ASA did not affect also the decrease in the bone blood flow after the administration of estradiol benzoate (EB). Local blood flow was determined by means of the uptake of 85Sr-microspheres. In experiment A (females) the local circulatory values were increased after OOX significantly in tibia and distal femur, insignificantly in diaphysis of the femur and calvaria; this increase after OOX was suppressed by simultaneous administration of ASA completely in tibia, partially (and statistically insignificantly) in distal femur and diaphysis of the femur; no effect of ASA could be demonstrated in the calvaria. The blood flow through the kidneys of OOX female rats was decreased after ASA, while no change occurred in muscle and skin. The bone mineral content was significantly lower in both groups of OOX females. In experiment B (males), we observed an increase in local circulatory values after ORX in tibia, distal femur and diaphysis of the femur and suppression of this increase by the administration of ASA. These changes were not demonstrable in the calvaria and soft tissues studied. In experiment C (females), we found that significant decrease in the local circulatory values in the tibia after 4 weeks of administration of EB was not altered by simultaneous administration of ASA. Experiment D (females) proved again that 24 hour incorporation of 45Ca and 3H proline was markedly suppressed by the administration of ASA not only in OOX females but also in sham operated control rats. Thus, the present results confirm the inhibitory effect of ASA on the increase in bone blood flow after castration, indicating with high probability participation of prostaglandin (PGE2?) (while no effect of ASA could be demonstrated on the decrease in bone blood flow after EB). The described blood flow changes are induced by local vascular reactions and occur in the bones of both female and male rats. Marked suppression by ASA of 45Ca and 3H-proline incorporation supports the circulatory results and indicates an important interconnection between metabolic processes and local circulation in the bone. We conclude that prostaglandin may be one of the factors regulating bone blood flow. PMID- 8711412 TI - [The Brothers of Mercy Hospital and Celestine Opitz]. AB - The Hospital of the Brethren of Mercy in Prague, Na Frantisku was built on the place, where a medieval hospital had been built in the middle of the 14th century. The Brethren of Mercy got into possession of the building in December 1620. Till the General hospital was built (1790) their hospital was the largest in Prague. Since the 2nd half of the 18th century the clinical education of the medicine students was held here. The Brother of Mercy Celestyn Opitz (1810-1866) was sent by the order to the Prague medical faculty, where he reached the magister of surgery in 1842. On February 7th, 1847 in the hospital Na Frantisku he was the first in Bohemia to apply the ether narcosis. In 1852 he graduated in Vienna to medicine doctor, but later he worked in the administration of his order. PMID- 8711413 TI - [Adolescentology--a new direction even in adolescent medicine?]. PMID- 8711414 TI - Cholesterol and atherosclerotic vascular disease: from science to public health policy. AB - Advances in our understanding of the pathology, epidemiology, genetics (particularly the recent research on transgenic animals) and the results of randomized prospective clinical trials all contribute to the formation of public health policy on serum cholesterol and other lipids. This article outlines the scientific background of these policies. There remains little doubt that dyslipidemia contributes to the development of coronary atherosclerosis and that effective treatment can prevent or retard its development. PMID- 8711415 TI - Interleukin-2-knockout mice: a new model to study autoimmunity and self tolerance. AB - Interleukin-2 deficient mice (IL-2-/-) generated by targeted mutagenesis proved to be an excellent system to study the function of this lymphokine in the context of whole immune system. In addition they provide an animal model of a primary immunodeficiency caused by a single-gene defect. Our results indicate that the essential defect in IL-2-/- mice is the lack of a negative regulatory function which leads to the break down of the self tolerance. The development of autoimmune diseases in IL-2 deficient mice stress the link between autoimmunity and primary immunodeficiencies. PMID- 8711416 TI - Metabolic interference of melanogenesis in pigment cells. AB - Biosynthesis of pigment melanin in pigment cells (melanocytes, nevus cells, melanoma cells) constitutes a distinctive but integrated part of intracellular metabolism. Although the melanin formation takes place in specialized compartments (melanosomes), these structures are not fully "leak-proof" and, as a consequence, the reaction products of melanogenesis have an opportunity to interact with different metabolic processes. This can be of particular importance when the melanogenesis is stimulated (e.g. by UV radiation) and when the integrity of melanosomes and/or their limited membranes is disturbed. From the toxicologic viewpoint melanogenesis represents an unique metabolic burden for the cell and this fact can have consequences for cellular defence systems. For instance, the conjugation of melanogenic precursors present in cytoplasm with glutathione or cysteine may reduce the availability of the thiocompounds for other necessary protective reactions. For the same reason, in melanin-producing cells there is an increased need for detoxification of the reactive intermediates by O-methylation. This reaction catalyzed by catechol-O-methyl-transferase may lead to concentration changes of methyl donor S-adenosylmethionine and the reaction product S-adenosylhomocysteine. As a consequence, decreased S adenosylmethionine/S-adenosylhomocysteine ratio (so-called "methylation ratio") is known to have profound inhibitory effect on methylation processes inclusive DNA methylation. Although still very speculative, this situation might very well lead to the generation of CpG mutations and (due to DNA hypomethylation) to activation of silent (onco)genes. PMID- 8711418 TI - Free radical activity of melanins and related substances: biochemical and pathobiochemical aspects. AB - Human body contains a number of natural pigments-authentic melanins, soluble melanins, neuromelanins, lipofuscins-having a similar origin-oxidation and polymerization of precursor phenols, and sharing many common characteristics among which dominates their free radical activity. Related substances can be formed from exogenous compounds introduced at some occasions to the organism. The functional properties of the pigments seem to be determined by their localization, i.e. cellular or extracellular, and as regards those intracellular ones by their subcellular compartmentalization. All the intracellular pigments mentioned in this study are associated with lysosomal enzymes at some stage of their development. A special attention was paid to a neglected group of soluble melanins. It was proposed that methylation may decide about the fate of precursor molecules whether they will join the soluble melanin pool or will be excreted as melanogens. Participation of the pigments in various biochemical and pathobiochemical processes has been reviewed. PMID- 8711417 TI - Melanogenic factors: regulation of gene expression. AB - Melanogenesis is a multistep biochemical process resulting in the formation of melanin in pigment cells in the skin and the eye. Three melanogenic factors, tyrosinase, TRP1, and TRP2 participate in the pathway. Here, the regulation of gene expression of these melanocyte-specific markers is shortly reviewed. PMID- 8711419 TI - "Epileptosis"--a syndrome or useless speculation? AB - 102 patients were divided into 3 groups: epileptics, psychotics and epileptics with psychotic symptoms. All had long been monitored for a number of clinical and laboratory parameters. Though different in many respects, all share states of sudden dysphoria, cacophoria, panic anxiety, horror, and EEG (stereo-EEG, too) signs of epileptic or other gross anomalies, often correlated to those affective disorders. Attacks of dysphoria, epilepsy, and psychosis come spontaneously and in response to biological (hypoglycemia, sleep deprivation, alcohol, menses) or psychosocial stimulation (agitation, quarrels, fear of redundancy, psychic trauma). These states (attacks, dysphoria, "neurotic" or even psychotic episodes) often provoke one another. -Calling this syndrome epileptosis, we believe its mechanism is due to lesions of the limbic and brainstem modulation systems. At the start of the process there is an epileptic focus in the amygdalo-hippocampal complex (AHC) which in itself can trigger simple or complex partial paroxysm but also-by means of electric stimulation of the AHC-states of dysphoria, anxiety, and psychotic hallucinations. Besides, a form of pathological learning develops in premorbid "hypersensitive" personality which can be put down to associative learning and to Overton's phenomenon of "state-dependent retention of learned responses". This may give rise to mutual stimulation where epileptic focal activity in AHC can provoke dysphoria while an external psychosocial situation can trigger epileptic activity there, too (AHC). Since there need not always be mydriasis (though other vegetative signs such as tachycardia, tachypnoea, nausea, blush and others are frequent) or unconsciousness, and some psychomotor manifestations may be out of the ordinary, and scalp EEG may be normal, such patients are often regarded as "hysterics" or malingerers. PMID- 8711420 TI - MESAM4 evaluated nocturnal respiration disturbances in myasthenia gravis. AB - Twenty-eight treated patients with myasthenia gravis (MG) (Osserman's scale: ocular form-12 cases, mild generalized form-5 cases, medium generalized form-10 cases, free of symptoms after thymectomy-1 case) were examined with MESAM4. Typical sleep apnea syndrome (SAS) patterns were observed in 12 subjects, mild saturation undulation without respiratory noises and without heart rate changes in 7 (3 of them also produced the SAS pattern) and irregular nonspecific changes in 3. The average vital capacity in SAS patients was 2556 (SD = 1166) ml, and in patients without MESAM4 abnormality 1629 (559) ml, P < 0.05. The average of oxygen saturation minima in SAS patients was 87.6 (3.6) %, in patients without MESAM4 abnormality 91.5 (1.8), P < 0.01. We conclude that there is no relation between any clinical parameter of MG and the occurrence of nocturnal respiratory disturbances. The risk factors of the occurrence of sleep respiratory disturbances in MG are similar to those of the non-myasthenia population although the incidence of this condition in MG is higher. PMID- 8711422 TI - High affinity serum-derived Fab fragments as another source of antibodies in the gut lumen of both neonates and adults. AB - The authors have investigated the presence of serum-derived immunoglobulin G (IgG) fragments in the human intestine at various ages, these fragments possibly representing another source of antibodies in addition to secretory IgA (SIgA). Fab fragments of the gamma isotype were found to be the major molecular form of immunoglobulins in the meconium (median value: 3.7 mg/g of stools), as compared with Fab alpha (75 micrograms/g) and IgM (2.6 micrograms/g). These fragments provided by molecules of the maternal serum displayed a strong antibody activity to the tetanus toxoid and were also found in the stools of 1-week-old babies fed with formula milk. The release of serum antibodies into the digestive lumen occurs largely via hepatobiliary secretions, as suggested by the presence of IgG antitoxins in the bile of children operated on extrahepatic biliary atresia. In adults, the Fab antitoxins were also detected in most stool extracts. Affinity of these molecules was found to be similar to that of their serum counterpart with a Ka of 2.1 x 10(10) M1 (median value). These mucosal antibody fragments, associated with the normal pathway of serum IgG catabolism, could provide additional immune defences against pathogens, and be of importance to supplement an immature or deficient secretory immune system. PMID- 8711421 TI - Complement subcomponent C1q modulation of TNF-alpha binding to L929 cells for enhanced TNF-mediated cytotoxicity. AB - Complement subcomponent C1q has been recently implicated in the modulation of autocrine binding of TNF-alpha to murine macrophages for induction of nitric oxide synthase. In the present study, the putative role of C1q in increasing TNF alpha binding to L929 cells to mediate cytotoxicity was explored. TNF-sensitive L929 cells (L929-S) had higher total endogenous cellular and surface C1q levels and bound correspondingly more phycoerythrin-labelled rTNF-alpha (PE-TNF) than did a TNF-resistant L929 variant (L929-R). Pretreatment of L929-S with soluble C1q increased their sensitivity to TNF-mediated cytotoxicity coincident with increased binding of PE-TNF, but similar treatment of L929-R had no effect. Pretreatment of L929-S with an inhibitor of C1q secretion, 3,4 dehydro-D,L proline (DHP), resulted in a decrease in their TNF-mediated cytotoxicity, as well as reduced binding of PE-TNF. Subsequent exposure of DHP-treated L929-S with exogenous soluble C1q restored their TNF-mediated cytotoxicity and binding of PE TNF. These results provide evidence for the modulation of TNF-alpha binding to TNF sensitive tumour targets L929 by either endogenously synthesized or exogenously added C1q to promote TNF-mediated cytotoxicity by mechanisms which remain to be elucidated. PMID- 8711423 TI - CD3-4-8- thymocyte precursors with interleukin-2 receptors differentiate phenotypically in coculture with thymic stromal cells. AB - CD3-4-8- interleukin-2 receptor positive (IL-2R+) thymocyte precursors from adult mice were cocultured with thymic stromal cells from syngeneic adult mice. The IL 2R+CD3-4-8- thymocytes were obtained by positive panning of IL-2R+ cells followed by either sorting or negative panning of triple negative cells, and they were cocultured with primary or secondary cultures of heterogeneous thymic stromal cells. Phenotypic maturation of these precursor cells was extremely rapid. Within 2 1/2 days significant numbers of CD4+8+ and CD3+4+8- cell populations developed, the latter expressing the alpha beta T-cell receptor (alpha beta-TCR). Thus heterogeneous stromal cell cultures support the development of IL-2R+ precursors and with these methods it will now be possible to isolate the particular stromal cells involved at each stromal-dependent step. PMID- 8711425 TI - Functional differences in the specific B-cell compartment in high or low antibody responder mice. AB - The role of antigen-presenting cells (APC) in quantitative antibody synthesis regulation was studied in mice genetically selected for high (HI) or low (LI) antibody response. Irradiated spleen cells and enriched specific B cells from HI and LI mice co-isogenic at H-2s locus, were compared for their capacity to present chicken ovalbumin (OVA) to specific T-cell hybridomas. Minor differences were observed between HI and LI mice when three distinct hybridomas were stimulated in the presence of OVA and splenic macrophages APC. These differences were totally abolished when APC were pulsed with OVAxAb complexes. Looking at the B-cell compartment, hybridoma IL-2 responses were similar when TNP primed B cells were pulsed with OVA. However, when OVA was targeted on TNP-specific enriched B cells by pulsing with TNP-OVA, the IL-2 production by the T-cell hybridomas was stronger in the presence of HI B cells than in the presence of LI B cells. These results strongly suggest that an efficient Ag handling/processing by specific B cells is a major component of the high Ab responder status in Biozzi mice. PMID- 8711424 TI - Detection of cruzipain, the major cysteine proteinase from Trypanosoma cruzi and its C-terminal extension in biological fluids during experimental infection in mice. AB - Monoclonal antibodies (MoAbs) specific for unique epitopes of the catalytic domain of cruzipain (Crz) were used to develop a two-site sandwich ELISA specific for native Crz. In addition, the authors developed a sandwich ELISA that allowed the detection of the protease C-terminal domain (CT) using a combination of a MoAb specific for the CT and rabbit anti-Crz IgGs. Both assays were sensitive with detection limits of 2 ng/ml and 0.7 ng/ml, respectively. The assays were assessed for applicability in detection of antigens in serum and urine from experimentally infected BALB/c mice. The antigens were already detectable in serum by the third week after infection, reached their peak by week four, and decreased during the chronic phase of the infection. Throughout the infection the relative amount of CT detected was several-fold higher than that of native Crz, and the data demonstrate that the cT exposes highly immunogenic epitopes that are absent in native Crz. Since these observations have a potential application in diagnosis, the authors analysed the degree of cross-reactivity with antigens from T. rangeli, T. brucei, Leishmania mexicana and L. panamensis, and determined that the assays were highly specific. Measurable amounts of the CT were also recorded in urine samples. PMID- 8711426 TI - Enhanced in vivo immunogenicity induced by an antibody to the IL-4 receptor associated gp200-MR6 molecule. AB - Four murine IgG1 monoclonal antibodies, each with specificity for a human tumour associated antigen, have been tested for their in vivo immunogenicity using a rabbit model. Surprisingly, one of these antibodies, MR6, was significantly more immunogenic than the remaining three reagents. This enhanced MR6 immunogenicity was not restricted to the immunoglobulin molecule itself, but also applied to a hapten (fluorescein isothiocyanate, FITC) when conjugated to the monoclonal antibody. In addition, the secondary immune response to an independent antigen, human haemoglobin, was higher when the antigen was administered simultaneously with MR6 than when co-injected with an isotype-matched control monoclonal antibody. The presence of the target antigen, gp200-MR6, on both rabbit and human leucocytes and epithelium, and its known association with human IL-4 function, raises the possibility that antibody MR6 may not only target immunogens to antigen-presenting cells, but may also enhance the ability of these cells to present antigen to the immune system. Antibodies to the gp200-MR6 may therefore find important clinical application as in vivo adjuvants. PMID- 8711427 TI - Monocyte and neutrophil adhesion to matrix proteins is selectively enhanced in the presence of inflammatory mediators. AB - The authors investigated the time course of monocyte and neutrophil adhesion to fibronectin, vitronectin and albumin precoated culture wells, using mixed leucocyte populations from healthy blood donors. Moreover, the influence of chemotactic agonists on the adhesion properties as well as the quantitative expression of CD29, CD11b/CD18 and CD61 was analysed by flow cytometry. Different chemotactic agonists were used representing a classical chemotactic agonist (fMLP), and agonists with a preferential effect on monocytes (RANTES) and neutrophils (IL-8), respectively. The authors found a gradual increase in monocyte and neutrophil adhesion to all three surfaces, reaching a plateau at 15 min of incubation. Adhesion to fibronectin was significantly higher at all time points (5, 15 and 60 min, respectively) compared with vitronectin and albumin in both monocytes and neutrophils. Neutrophil adhesion to vitronectin was significantly lower at 60 min compared with 15 min. Monocyte adhesion to albumin was increased by fMLP and RANTES and to vitronectin also by IL-8. Neutrophil adhesion to albumin and vitronectin was increased by fMLP and IL-8, but not RANTES. The adhesion to fibronectin was not altered by any of the chemotactic agonists used. The quantitative levels of CD11b/CD18, but not CD29 and CD61, was increased by fMLP, but not RANTES nor IL-8. The authors conclude that the adhesion of human monocytes and neutrophils to vitronectin and albumin, but not fibronectin, is selectively enhanced by chemotactic agonists and may contribute to the selective accumulation of different leucocyte subsets at the inflammatory site. PMID- 8711428 TI - Differential effect of the activation of protein kinase A on the protein synthesis and secretion in the T-helper 2 cell line D10.G4.1. AB - The authors analysed the effect of protein kinase A (PKA) activation on the protein synthesis and secretion in the T-helper 2 cell line D10.G4.1 (D10) using an assay that allows the detection of almost all secreted proteins of a cell. IL 4 and IL-10 were quantified. Three groups of secretory products could be defined. The T-cell receptor (TCR)-induced production of the first group (A) of proteins including IL-4 was enhanced by low concentrations of PKA activators. At higher concentrations the enhancement was less marked. The synthesis and secretion of a second group (B) of proteins including IL-10 remained unaffected. The production of a third group (C) of proteins was inhibited in a concentration-dependent manner. Biochemical analysis revealed a block of phospholipase C gamma (PLC gamma) activity by PKA activators. When D10 cells were stimulated by a phorbol ester plus calcium ionophore the production of group A proteins was enhanced almost fourfold, whereas production of group B proteins was unaffected by PKA activation. This effect was observed at all concentrations of various PKA activators tested. The secretion of group C proteins was no longer inhibited. The same results were obtained when analysing IL-4 and IL-10 m-RNA by Northern blotting. The data demonstrate a lymphokine specific mode of action on a single cell basis. Furthermore, it suggests that the inhibitory action of PKA in D10 cells is due partly to blocking of PLC gamma activity. PMID- 8711429 TI - Normal human immune peritoneal cells: subpopulations and functional characteristics. AB - Normal human peritoneal cells collected during elective laparatomy from patients with gallbladder stones without clinically detectable inflammatory changes were characterized phenotypically with immunocytochemical method and flow cytometry, with special attention paid to the presence of memory cells. The responsiveness of normal PCs to mitogen and, specifically, the role of peritoneal macrophages in this process was studied. The peritoneal cells consisted of 45% of monocytes/ macrophages (CD68+), as many as CD2+ T lymphocytes, 8% CD57+ NK and K 2% CD22+ B, cells. The CD4/CD8 ratio was 0.4. The peritoneal cells did not express interleukin-2 (CD25+) and transferrin receptors (CD71+) on their surface. Approximately 49% of the peritoneal cells were class II MHC antigen positive cells. Two per cent of S100+ dendritic cells were found. Flow cytometric two colour analysis revealed that the majority of peritoneal CD4+ (92.4%) and CD8+ (73.1%) lymphocytes, while only 50.2% of CD4+ and 30.1% CD8+ peripheral blood cells expressed simultaneously the CD45R0 (UCHL1) molecule, which is characteristic to the memory/effector T-cell subpopulation. Peritoneal T lymphocytes responded to the mitogens less than peripheral blood lymphocytes of the same individual. Supplementation of cell culture with anti-macrophage (anti CD68) and anti-HLA-DR MoAb brought about a dose-dependent decrease of proliferative peritoneal cell response to ConA. The authors conclude that human peritoneal cell population comprises a high proportion of T lymphocytes and macrophages capable of presenting antigens to peritoneal lymphocytes. High prevalence of memory lymphocytes points to the preparedness of these cells to react with invading antigens most likely of gut bacterial origin. PMID- 8711430 TI - The cell surface phenotype of human natural interferon-alpha producing cells as determined by flow cytometry. AB - The relationship between the so-called natural interferon-alpha (IFN-alpha) producing cell (IPC), stimulated to produce IFN-alpha by herpes simplex virus type 1 (HSV), and of dendritic cells (DC) in peripheral blood leucocytes was investigated. The simultaneous expression of cell surface antigens and intracellular IFN-alpha in the HSV-stimulated IPC (HSV-IPC) was examined by flow cytometry (FCM). The HSV-IPC were infrequent, < 0.3% of the mononuclear leucocytes, and with homogeneous light scatter characteristics. The HSV-IPC were confirmed to lack leucocyte lineage specific markers, and to express CD4, CD36 and HLA-DR. Furthermore, they expressed high levels of CD44, CD45RA and CD45RB, and lower levels of CD40, CD45R0, CD72 and CD83. The HSV-IPC expression of CD13, CD33 and Fc epsilon RI were weak but significant, while no CD5, CD11b, CD16, CD64, CD80 or CD86 were detected. Sorted pure HSV-IPC had irregular shaped nuclei, many mitochondria and vesicles, and rugged cell membranes without veils. Sorted HSV-IPC stimulated proliferation of autologous T cells from HSV immune donors. Thus, the HSV-IPC in many respects resemble immature DC, but clearly differ from typical mature DC. However, they may also represent a specialized population of efficient IFN-alpha producing cells. PMID- 8711431 TI - Genetic influence on the shaping of the human T-cell receptor repertoire: quantitative assessment by competitive polymerase chain reaction. AB - It has been difficult to define the different factors which contribute to the shaping of the human T-cell receptor (TCR) repertoire. In this study, the influence of the polymorphic human leucocyte antigen (HLA) genes and non-HLA genes on the phenotype of the TCRBV segment repertoire was assessed in a population of HLA class I-matched individuals including three pairs of siblings. The gene expression levels of 24 TCRBV families were evaluated in the CD4+ and CD8+ T-cell subsets of unstimulated peripheral blood mononuclear cells (PBMC) by reverse transcription (RT) and a newly developed competitive polymerase chain reaction (cPCR) assay. Titration experiments demonstrated that the RT-cPCR assay was suitable for an accurate quantification of the relative TCRBV segment expression levels. The T-cell repertoires of HLA-identical siblings were found to be more similar than the repertoires of unrelated individuals. On the other hand, there was no difference in the degree of similarity between the TCRBV repertoires of CD4+ T-cells of HLA class II identical or non-identical unrelated individuals. Furthermore, although most of these individuals had identical HLA class I genes and non-identical HLA class II genes, the TCRBV repertoires of the CD4+ T cells exhibited significantly lower variabilities than the repertoires of the CD8+ T cells. The results of the RT-cPCR assay were supported by flow cytometric analysis of the CD4+ and CD8+ T-cell subsets of the same eight individuals employing 10 different TCRBV segment-specific monoclonal antibodies. These observations argue for a predominant role of non-HLA or non-polymorphic HLA determinants for the shaping of the TCRBV repertoire. PMID- 8711432 TI - Strict adherence to a common rank order of T-cell receptor V beta usage in human leucocyte antigen disparate individuals. AB - In order to investigate the T-cell receptor (TCR) V beta usage in different T cell subsets, the authors performed flow cytometric analyses using a large panel of TCR V beta-specific monoclonal antibodies on CD4+, CD8+ CD28+ and CD8+ CD28- T cells from 15 random blood donors, six umbilical cords and seven human leucocyte antigen (HLA) identical non-twin sibling pairs. The authors found that the proportion of T cells expressing each V beta gene product was similar within CD4+ and CD8+ CD28+ T cells from all samples studied. For these T-cell subsets a rank order of V beta usage could be identified which was adhered to by all donors. In contrast, within CD8+ CD28- T cells a wide variation of V beta usage was found between individuals, and no rank order correlation could be detected. Members of HLA identical sibling pairs were found to be no more similar in their usage of V beta gene products than pairs of HLA disparate random blood donors. Groups of individuals sharing HLA antigens were no different from the groups not sharing such antigens in their usage of V beta segments. The results suggest that HLA polymorphisms play no more than a minor part in determining TCR V beta usage. PMID- 8711433 TI - Activation of the complement, coagulation, fibrinolytic and kallikrein-kinin systems during attacks of hereditary angioedema. AB - Five patients with hereditary angioedema (HAE) were studied during attacks and remission as were healthy controls. The high levels of C1/C1-INH complexes, low C4 and high ratio C4 activation products (C4bc)/C4 also differed significantly during remission compared to controls. During attacks C4bc/C4 increased (922 2007; P = 0.022, remission versus attacks, median values throughout), C2 and CH50 dropped (111-31%; P = 0.043 and 110-36%; P = 0.016, respectively), TCC (C5b-9) increased (0.88-1.23 AU/ml; P = 0.028). Cleavage of HK increased to be almost complete during attacks (20-90%; P = 0.009). While factor XIa/serpin-complexes did not increase, a more than twofold rise in thrombin/antithrombin-complexes (0.20-0.50 microgram/l; P = 0.009) and in plasmin/alpha-2-antiplasmin-complexes (7.3-17 nmol/l; P = 0.028) was observed. For the first time cascade activation in HAE was studied simultaneously, and corroborates that attacks lead to activation of the kallikrein-kinin system, fibrinolysis and early part of the classical complement pathway. In addition, the authors present novel data of terminal complement and coagulation activation, the latter apparently not via FXIa. PMID- 8711434 TI - Histaminergic regulation of NK-cells: protection against monocyte-induced apoptosis. AB - Human natural killer (NK) cells (with CD3-/56+ phenotype) acquired features characteristic of apoptosis after incubation with autologous monocytes, as revealed by apoptotic nuclear morphology and degradation of DNA into oligonucleosomal fragments. The monocyte-induced apoptosis in NK-cells was prevented by the biogenic amine histamine at concentrations exceeding 0.1 microM. The protective effect of histamine was blocked by the H2-receptor (H2R) antagonist ranitidine but not by AH202399 A, a chemical control to ranitidine devoid of H2R affinity. It is concluded that histaminergic mechanisms may serve to protect NK cells from damage inflicted by products of the oxidative metabolism of monocytes. PMID- 8711435 TI - Trypanosoma cruzi cytosolic alkaline antigens (FI) induce polyclonal activation in murine normal B cells. AB - Several reports have described polyclonal activation in mice acutely infected with Trypanosoma cruzi. The aim of this work was to analyse the participation of one T. cruzi antigenic fraction in this immunological event. The antigen selected was FI, an antigenic fraction of pI 7-9 obtained from T. cruzi cytosol separated by isoelectricfocusing. FI is constituted by molecules with molecular weights of around 60 and 20 KDa. The authors assayed the ability of this antigenic fraction to induce polyclonal activation of spleen mononuclear cells from normal (NSMC) BALB/c mice. NSMC showed a marked lymphoproliferative response measured by 3H thymidine incorporation after 3 days of culture in presence of FI. The values reached by FI-stimulated cells were 10 times higher than the controls (non stimulated cells). This effect was dose-dependent. Furthermore, the authors observed that a purified T-cell population in the presence of adherent cells was unaffected by FI. Additionally, in a culture of NSMC, FI stimulated the proliferation of B cells as observed by the increase of the percentage of B220+ cells determined by FACS using FITC-conjugated anti-mouse B220. The authors noticed that the percentage of B220+Ly1+(CD5) populations in the presence of FI did not change with respect to the control (non-stimulated cells), indicating that FI expanded both conventional and CD5+ B cells. The isotypic pattern of the antibodies produced after 6 days of culture of NSMC in the presence of FI was predominantly IgM, which reacted with highly conserved antigens such as actin, myosin, myoglobin, thyroglobulin and carbonic anhydrase, but did not react with FI. A slight increase of IgG1 and IgG3 with respect to the control was observed but no changes on the levels of IgG2 was noticed. These results indicate that FI promotes activation, proliferation and differentiation in antibody-secreting cells of normal murine B lymphocytes. PMID- 8711436 TI - Loss of neurons in the dorsal root ganglia after transection of a peripheral sensory nerve. An anatomical study in monkeys. AB - Injury to a sensory nerve often results in a poor long term outcome, partly because of sensory motor mismatch of regenerating axons at the transection site. We studied nine macaque monkeys and found that 27% of nerve cells in the projecting dorsal root ganglia had been lost 21 months after transection and suturing of the radial sensory nerve. No specific cell sizes were lost and the reduction was evenly distributed in the affected ganglia in which neurons had been labelled with a mixture of wheat germ agglutinin-horseradish peroxidase conjugate (WGA-HRP) and HRP alone. PMID- 8711437 TI - Effect of beraprost sodium (Procylin), a stable prostaglandin I2 analogue, on a dorsal skin flap model in rats. AB - Beraprost sodium, a chemically stable prostaglandin I2 analogue with antiplatelet vasodilator actions, was tested by comparing the extent of viability of random pattern skin flaps in control rats and rats treated with beraprost sodium. An isolated tail based 2 x 7 cm flap was constructed in 50 Sprague-Dawley rats. In group 1 (beraprost sodium treated group, n = 25) 0.015 mg beraprost sodium were injected intraperitoneally immediately after the flap had been raised. In group 2 (control group, n = 25) the animals were injected intraperitoneally with the same volume of saline immediately after the flap had been raised. At the end of the seventh day the flap survival area was documented using digital planimetry. The difference between the results showed that beraprost sodium had significantly beneficial effect on skin flap survival (p < 0.01, Welch's test). PMID- 8711438 TI - Effects of lipo-prostaglandin E1 on random pattern flaps in rabbits. AB - The effects of lipo-prostaglandin (PG) E1, which are lipid microspheres containing PGE1, were examined using 2 x 12 cm random pattern skin flaps on the backs of five rabbits. Five rabbits acted as controls. Both surviving length (8.5 (3.1) compared with 4.9 (1.8) cm) and surviving area (22.1 (9.3) compared with 11.5 (6.1) cm2) of the skin flap were significantly increased (p < 0.03 in each case). In three additional rabbits the blood flow on the distal side of the skin flap, where it was poor, was measured using a laser image blood flowmeter. Blood flow was improved five to 15 minutes after injection of lipo-PGE1 and was maintained for at least 60 minutes. In conclusion, lipo-PGE1 increased the blood flow of the random skin flaps, resulting in significant enlargement of the surviving area of skin flaps. PMID- 8711439 TI - Reduction mammaplasty with adrenaline infiltration: effects on perioperative bleeding. AB - We present a retrospective comparison of two groups of patients who underwent reduction mammaplasty with (n = 200) and without (n = 44) infiltration of a combination of a local anaesthetic and adrenaline. The vasoconstrictive effect of adrenaline reduced the intraoperative bleeding from 0.5 ml to 0.1 ml/g resected tissue. Postoperative bleeding was also reduced. Drains were omitted in 104 of the patients who had received adrenaline. In spite of this, these patients had no increases in the rates of infection, haematoma formation, or need for reoperation compared with the group that had been given drains. The practical implication is that after local infiltration of a vasoconstrictor agent post-operative drains can be omitted, at least in the Strombeck reduction mammaplasty procedure. PMID- 8711440 TI - Quality of the capsule in reconstructions with textured or smooth silicone implants after mastectomy. AB - Many women desire some kind of reconstruction after mastectomy for breast cancer to improve the cosmetic result. Since the introduction of silicone breast implants, capsular contracture has been the most common complication of breast augmentation or reconstruction. The aim of the present study was to evaluate the capsule quality in patients with breast cancer who had undergone reconstructions with either textured (rough) or smooth silicone implants after simple or modified radical mastectomy. Forty-three consecutive patients had reconstructions with textured silicone implants (49 implants) and were evaluated 32 months post operatively in a prospective clinical trial. The last 43 consecutive patients who had reconstructions with bilumen smooth silicone implants (49 implants) acted as controls. The results were graded by Baker's classification. Among the patients with textured implants only 9% (4/47) was graded as having moderate or severe capsular contracture in contrast to 24% (11/46) in patients with bilumen smooth surface implants (p < 0.05). These results confirm previous reports that favour textured silicone implants in breast augmentation and reconstruction. PMID- 8711441 TI - Emergency free flap cover in complex injuries of the lower extremities. AB - Nineteen patients with complex injuries of the lower extremities were treated with emergency free flap reconstruction after radical debridement, bone stabilisation, and primary repair of injured structures (nerves, tendons, vessels, and muscles). Two patients sustained complete below knee amputation associated with avulsion of soft tissue and had salvage free flap transfer to preserve length of stump and function of knee joint. Long-term follow up ranged from four months to eight years (median 3.5 years) No flaps failed and there were no serious complications of wound healing. Procedures were associated with minimal morbidity, a reduced number of secondary operations, shorter hospital stay, and good functional results. PMID- 8711443 TI - Donor site morbidity of radial forearm flaps. A clinical and ultrasonographic evaluation. AB - Harvesting of a forearm flap based on the radial artery has been thought to cause functional or circulatory problems in the donor hand. Eighteen patients were examined three to 24 months after a radial forearm flap had been raised. The function of both hands was studied for grip strength, mobility of the wrist and elbow joints, and sensitivity of the area served by the superficial radial nerve. The patients were interviewed and the cosmetic result was evaluated. Duplex ultrasonography and colour Doppler ultrasonography of both ulnar arteries were done, and the brachial arteries were measured as controls. Angle-corrected peak flow velocity (cm/s) in the ulnar artery of the donor forearm was significantly increased at the level of the wrist compared with the control forearm (100.9 compared with 73.1 cm/s, p = 0.017), as was the ulnar: brachial peak flow velocity ratio (1.18 compared with 0.76, p = 0.001). The grip strength of the donor hand was weaker by 11.9% (86.5 compared with 72.2 Kp), 10 (56%) had areas of sensory loss over the radial nerve distribution, and seven of the 18 patients complained of cold intolerance. Four patients considered the donor site result so bad that they would not have chosen the operation had they known what the result would look like. The radial forearm flap donor site is not without problems, and the patients must be carefully selected and properly informed preoperatively. PMID- 8711442 TI - Extended free lateral arm flap with preservation of the posterior cutaneous nerve of the forearm. AB - The free lateral arm flap has become a well-defined and reliable flap for various reconstructive purposes. Little attention has been paid, however, to the possibility of preservation of the posterior cutaneous nerve of the forearm (nervus cutaneus antebrachii posterior) sacrifice of which results in numbness of the dorsal part of the forearm. In this study, an anatomical dissection showed that in many cases it would be possible to preserve the nerve. We did 23 free lateral arm flaps in 22 patients during the period 1989-1994. The maximum flap length was 40 cm. Standard maximum width in most cases was 6 cm, and by using a new expansion technique it reached 10 cm in one case. Furthermore, with meticulous dissection the posterior cutaneous nerve of the forearm was either preserved or cut and rejoined in 21 patients, so minimising sensory loss at the donor site. PMID- 8711444 TI - Cold-induced vasospasm after finger replantation; abnormal sensory regeneration and sensitisation of cold nociceptors. AB - Cold-induced vasospasm which may present clinically as "white fingers", after hand injuries has been reported to be as high as 100% after replanted digital amputations. The exact cause of this is obscure and no specific treatment is available. To try to shed light on the cause of post-traumatic cold-induced vasospasm we evaluated replanted digits in seven patients who had had replantations more than 10 years ago. Our results show that cold-induced vasospasm occurred in six out of seven patients. Cold nociceptors were sensitised in patients who had abnormal two point discrimination, all of whom responded to cooling by vasospasm. One patient with normal circulation did not recover cold nociception during cooling. This indicates that secondary Raynaud's syndrome after injuries to the hand may be related to sensitisation of cold nociceptors. Even simple nerve injuries may lead to secondary Raynaud's syndrome. PMID- 8711445 TI - Osteotomy of the first metacarpal for osteoarthrosis of the basal joints of the thumb. AB - A radial, 30 degrees wedge osteotomy was used for treatment of osteoarthrosis of the trapeziometacarpal joint in 18 patients, 16 women and two men, mean age of 61 years (range 38-74). Six of them had involvement of the scaphotrapezial joint. After a mean follow-up of 8.5 months (range 2-21) one of the patients were completely satisfied with the result of the operation, seven were quite satisfied, and two considered themselves unchanged. None was made worse. Of the 12 patients who did not have involvement of the scaphotrapezial joint nine were completely satisfied and three quite satisfied. Grip strength on the operated side was 75.5% (32%-100%), and pulp pinch was 74.5% (30%-100%) of that in the contralateral one. The mobility of the thumb was influenced little by the operation. Wedge osteotomy of the first metacarpal is a simple and safe way of relieving pain, particularly in the early stages of osteoarthrosis of the basal thumb joint. PMID- 8711446 TI - Regeneration across a partial defect in rat sciatic nerve encased in a silicone chamber. AB - Forty-four rat sciatic nerves with partial defects were repaired with a silicone chamber. Each partial defect was created by resecting a 10 mm segment from the tibial fascicle leaving the peroneal fascicle intact. The proximal and the distal stumps of the tibial fascicle together with the intact peroneal fascicle were encased in a single silicone chamber. After seven days a fibrin matrix had surrounded the peroneal fascicle and spanned the defect between the tibial stumps. This matrix was later invaded by non-neuronal cells and regenerating axons. Non-myelinated nerve fibres had almost regenerated across the defect by 16 days. The tetanic force of the gastrocnemius muscle 120 days after repair showed 80% recovery, which was no different from that of partial defects repaired with conventional nerve grafts. The results suggest that the silicone chamber technique could be applicable to the treatment of partially transected nerve trunks. PMID- 8711447 TI - Camptodactyly caused by an anomalous origin of the flexor digitorum superficialis tendon. Case Report. AB - A 20 year old man with a progressive flexion contracture of the proximal interphalangeal joint of his right ring finger had an anomalous origin of the flexor digitorum superficialis tendon. After excision of the aberrant flexor tendon the patient had a normal range of movement of the proximal interphalangeal joint. PMID- 8711448 TI - Fibrohistiocytic sarcoma of the thumb treated by wide resection and immediate free flap reconstruction. Case report. AB - A 29 year old surgical resident presented with a fibrohistiocytic sarcoma over the thumb of his dominant right hand. The tumour was widely excised including the ulnar neurovascular bundle. The resultant defect was immediately reconstructed with a sensory forearm free flap from the opposite upper extremity. The patient was then given a curative dose of cobalt-60 radiotherapy starting in the fourth postoperative week. With a comprehensive regimen of physiotherapy and splinting, the patient regained an almost full range of movement in his right thumb and was able to resume his regular duties in the operating theatre two months after his operation. Soft tissue sarcomas of the hand are a real therapeutic challenge. Adequate control of the tumour in most instances requires amputation because limb sparing procedures require soft tissue coverage not available locally or achieved easily. In addition, preservation of a hand, or parts of it, with poor functional capacity, is not helpful. PMID- 8711450 TI - Predicting 13-year-olds' drinking using parents' self-reported alcohol use and restrictiveness compared with offspring's perception. AB - This study examined whether offspring's perception of parental frequency of alcohol use and restrictiveness towards offspring's alcohol use could be used as a substitute for parental self-reports. Offspring's perception were compared with parents' self-reported alcohol use and restrictiveness in ability to predict offspring's own alcohol use. Respondents were 924 offspring, 642 fathers and 729 mothers. Correlations between fathers' and mothers' self-reported frequency of drinking alcohol and offspring's report of parental frequency of drinking alcohol ranged from 0.55 to 0.70. Fathers' and mothers' self-reported alcohol use and restrictiveness explained less than two per cent of the variance in their offspring's alcohol use in regression analyses, while offspring's perception of parental drinking frequency and restrictiveness explained about eight per cent of the variance in offspring's alcohol use. These results represent a challenge to studies of young adolescents which interpret offspring's reports of parents' alcohol use as a surrogate measure of parents' self-reported alcohol use. PMID- 8711451 TI - Spectral factors in the perception of vowel quantity in Icelandic. AB - Previous research has shown that the ratio of vowel to rhyme (vowel + consonant) duration is a major cue for quantity in Icelandic. In particular it serves as a higher-order invariant which enables the listener to disentangle those durational transformations of the speech signal which are "extrinsic" (e.g. due to changes in speaking rate) from those which are "intrinsic" to the phonemic message, involving a change of phonemic quantity. Previous research has been based on speech segment contrasts which are purely durational, involving vowels with a uniform spectrum whether phonemically long or short, such as [a] or [I]. This paper looks at the role of spectral factors in vowels which are spectrally dissimilar in their long and short varieties. It is shown that in these cases the spectral differences can be sufficiently great to override the previously established relational invariant for quantity. The implications of this finding for a model of quantity perception are discussed. PMID- 8711449 TI - Polydactyly. PMID- 8711452 TI - Speech gestures and facial expression in speechreading. AB - Two experiments investigated the role of facial expression in speechreading. In Experiment 1 sentences were presented either with only the actor's face shown or with the actor shown in half-size. One happy script with happy, sad and neutral sentences was used, and either a content cue or an emotional cue was presented prior to each sentence. In Experiment 2 happy and sad sentences from one happy and one sad script were first recorded by the actor with a neutral face (face and shoulder shown) and then with a congruent facial expression. The results revealed that there was no difference in performance between half-size and face presentation, but content cues increased performance compared to emotional cues. As expected (Abelson, 1981), happy sentences were easier to speechread than sad in the happy script and vice versa. Also, facial expression raised performance for happy sentences in the happy script. Positive emotions are suggested to be associated with approach behavior in part relying on increased non-pantomimic speech gestures and articulation of the speaker (e.g., Davidson et al., 1990). PMID- 8711454 TI - Developmental pathways of behavior problems in the young child: factors associated with continuity and change. AB - The continuity and discontinuity of behavioral problems from age 4 through age 7 until 10 were examined in 120 children. The mothers rated childhood behavior problems at the three age points. Problems were defined as above 80th percentile. Individual transition classification of problematic and non-problematic children showed that 7% had stable behavior problems, 60% stable non-problematic behavior and the rest displayed transitions related to either onset or offset of problems. The mechanisms involved in onset were different from those involved in offset. Children showing onset of problems had fewer friends, participated less often in organized leisure activities and were more passive in class than the well functioning children. Maternal characteristics such as continued education, current employment and high educational ambitions for the child were associated with having children changing their pathway from risk to an adaptive one. PMID- 8711453 TI - Teaching reading to disabled readers with language disorders: a controlled evaluation of synthetic speech feedback. AB - In a long-term study two groups of language and reading impaired students (N = 15 + 15) were reading with the aid of segmented speech-feedback in a computerized program. One group received feedback that was simultaneously segmented visually and auditorily into syllables, the other received feedback by letter names. In both groups subjects were expected to synthesize segments into words and to compare their synthesis to whole word feed-back subsequently provided by the computer. They worked for half a lesson (approximately 20 minutes) a day for a total of 40 days. During this period the experiment groups progressed more in reading than a control group of age and reading-level-matched students (N = 35) who received traditional remedial instruction. The group in the syllable condition gained slightly more in non-word reading and in syllable segmentation than did the letter group. Differences in gains in reading abilities were not explained by differences in age, but to some extent by initial level of phoneme and syllable awareness. Future applications of the speech-feedback system are discussed. PMID- 8711455 TI - The empirical relationships between standardized measures of religiosity and personality/mental health. AB - The aim of this study was to investigate the relationships between standardized, factor-based measures of religiosity and personality/mental health. In a sample of 471 self-identified Christian subjects, 303 females and 168 males, 79 non psychotic psychiatric in-patients and 392 non-patients, personal extrinsicness was partially positively correlated with the BCI Obsessive score. In multiple regression analyses some of the factor-based religious orientation indices related differently to the BCI Oral, Obsessive and Hysterical Scales and the SCL 90 Global Symptom Index as dependent variables. The religious orientations explained 8.8% of the variance of the BCI Oral Score, 4.2% of the BCI Obsessive score, 3.3% of the BCI Hysterical score, and 12.3% of the SCL-90 Global Symptom Index score. Of the doctrinal belief and morality indices only with Moral conservatism was significantly related to the BCI Hysterical score, and then negatively. PMID- 8711456 TI - Psychometric properties of the Danish 16PF and EPQ. AB - A Danish translation of the Cattell's 16PF has been used in studies evaluating the effects of prenatal drug exposure. This paper reports a psychometric analysis of the 16PF and Eysenck's EPQ based on a sample of 558 young Danes. Many 16PF scales had unacceptable psychometric properties (as indicated by coefficient alpha and item--total score correlations), but more satisfactory results were obtained with the EPQ N and E scales. A factor analysis of all 16PF and EPQ scales suggested a six factor solution that roughly corresponds to the second order factor structure obtained by Krug and Johns (1986). It is concluded that the second-order factor structure should be the basis of interpretation of the 16PF in both practical and research contexts. PMID- 8711457 TI - [Serious plant poisonings in Switzerland 1966-1994. Case analysis from the Swiss Toxicology Information Center]. AB - AIM: To analyze the types, frequency and severity of plant poisonings in Switzerland over 29 years. METHODS: Retrospective analysis of severe poisonings with toxic plants reported to the Swiss Toxicological Information Center (STIC). Assessment of the causality, severity of symptoms and the types of plants involved. RESULTS: During the period under review the STIC registered 24 950 cases of contact with or ingestion of toxic plant material. In 99.4% of all cases the clinical course was either unknown, asymptomatic or associated with only minor symptoms (no hospitalization). Severe plant poisonings occurred in 152 cases. Detailed analysis was possible in 135 cases (23 children, 112 adults) including 5 lethal cases (all adults). The 24 plants involved produced the following severe symptoms: Atropa belladonna (42 cases): anticholinergic syndrome (42), acute psychosis (33), convulsions (2), coma (2). Heracleum mantegazzianum (18): severe photodermatitis (18). Datura stramonium (17): anticholinergic syndrome (17), psychosis (12), coma (2). Dieffenbachia (11): severe stomatitis (8), corneal lesions (3). Colchicum autumnala (10): diarrhea (10), liver necrosis (9), fatal multiorgan failure (2). Veratrum album (8): bradycardia ( < or = 40/min) (6), shock (5). Aconitum napellus (4): tachyarrhythmia (2), AV-block II/III (2). Aesculus hippocastanum (3): allergy (3), anaphylactic shock (2). Hyoscyamus niger (3): anticholinergic syndrome (3). Ricinus communis (3): diarrhea (3), toxic megacolon (1). Oenanthe crocata (2): convulsions (1), lethal coma (1). Taxus baccata (2): tachyarrhythmia (1), fatal asystole (1). Further single cases with severe poisonings were observed with Arum maculatum, Asarum europaeum, Chrysanthemum vulgare, Cyclamen persicum, Datura suaveolens, Glycyrrhiza glabra, Laburnum anagyroides, Lycopodium, Narcissus pseudonarcissus (lethal aspiration), Nerium oleander, Senecio vulgaris and Vicia faba. CONCLUSIONS: Potential and real intoxications with plant materials occurred in 7.2% of all cases registered at the STIC. However, among all plant cases only 0.6% were severe intoxications requiring hospitalization. Although severe plant intoxications are rare events, a small number of specific plants appear to be mainly responsible for continued serious plant poisoning in Switzerland. The present study has identified the plants with the highest toxicological risks and provides a data base for more rational prevention, diagnosis and treatment of plant poisoning cases in the future. PMID- 8711458 TI - [Trends in alcohol consumption in Switzerland from 1975-1992]. AB - Based on four representative surveys independently conducted at 6-year intervals, the study presented here delineates the development of alcohol consumption in the general population of Switzerland between 1975 and 1992. A description of changes in the frequency and quantity of alcohol consumption by sex is presented, as well as specific changes by beverage type. Even though Switzerland remains one of the high consumption countries, a marked decline can be observed. Compared to 1975 when 28% of respondents reported daily alcohol consumption, this figure dropped to 20% in 1992. Although 10% consumed more than 60 grams of pure alcohol in the 1975 sample, this proportion fell to only 3% in 1992. This downward trend, which is also reflected in secondary statistics such as sales and mortality figures, can be mainly attributed to male consumers. In contrast to international trends, which in general demonstrate converging drinking patterns, reduced consumption in Switzerland, a wine-producing country, is mainly due to a decline in beer consumption and not in wine consumption. PMID- 8711459 TI - [Paraplegia following intrathecal chemotherapy]. AB - The occurrence of major neurologic disturbances is a rare complication of intrathecal cytotoxic drug administration. We describe the case of a 30-year old man with relapsed lymphoblastic lymphoma and CNS involvement who was treated by systemic chemotherapy (ESHAP) and intrathecal injections of methotrexate, cytarabine, thiotepa and hydrocortisone. Thereafter he developed persistent paraplegia with sensory and sphincteric insufficiency. The role of the drug in causing this syndrome was suspected when more common causes, such as a meningeal carcinomatosis, had been excluded by means of CNF analysis and radiological examinations. We review this and other published cases and discuss the possible pathogenesis, as well as the clinical and paraclinical findings in paraplegia following intrathecal chemotherapy. Patients particularly at risk are those who receive intrathecal chemotherapy for over CNS disease rather than as prophylaxis, those who receive several injections and patients who are concomitantly treated with radiotherapy to the brain or systemic high-dose methotrexate or cytarabine. PMID- 8711460 TI - [Tuberculosis in Switzerland in 1996: prevention and treatment]. AB - Tuberculosis is currently a rare disease in Switzerland and its incidence is slowly decreasing. Nevertheless, the increase in the proportion of foreign-born patients, the occurrence of multidrug-resistant bacilli and the increase of tuberculosis in HIV-infected individuals give rise to concern and have prompted changes in the management of the disease. The first aim is to cure the patients with adequate chemotherapy in order to prevent relapses and the development of drug-resistant germs. The best way to do so is to provide directly supervised treatment, particularly in cases where compliance with the treatment may be uncertain. Tuberculosis is transmissible to sensitive individuals. Therefore, some preventive measures are still indicated in settings where patients with a known or unknown bacillary form of tuberculosis may be cared for. PMID- 8711461 TI - Surveillance of influenza in Switzerland between 1987 and 1995. AB - During 8 years of continuous influenza surveillance in Switzerland (1987 to 1995), influenza A viruses predominated during 5 seasons and influenza B viruses during 3. The most severe outbreaks occurred in the 1988/89 season (A/H1N1 subtype), in the 1989/90 season (A/H3N2 subtype) and in the 1994/95 season (simultaneous outbreak of influenza A/H3N2 and B). From 1987 to 1993, peak activity of influenza A viruses was observed during December and January (for 8 weeks on average) while influenza B viruses were most active between February and March (ordinarily for 6 weeks). During the 1994/95 season, however, simultaneously increased activity of both influenza A/H3N2 and B viruses was observed, peaking at the end of March and lasting for about 9 weeks. The variants predominant in Switzerland largely corresponded antigenically to those isolated elsewhere in Europe, but a delay of one year was often observed. New strains detected at the end of a season (herald viruses) made it possible to forecast the type or subtype of virus which would predominate the next season. Isolation rates for A/H3N2 viruses were significantly higher in the 10-19 years and 60-plus age groups, whereas there was no statistical difference between age groups for A/H1N1 viruses. A decreasing isolation rate, corresponding to increasing age, was found for influenza B viruses. Increased mortality was observed in patients over 65 during the period of high influenza A/H3N2 activity, as was especially evident during the 1989/90 season. PMID- 8711462 TI - [Catheter closure in atrial septal defects and open foramen ovale using the Sideris buttoned device]. AB - Percutaneous nonsurgical closure is an alternative to surgical repair of a significant atrial septal defect (ASD) or a patent foramen ovale (PFO) associated with systemic embolism without other cause. We report the results of the first 35 patients (19 ASDs, 16 PFOs) in whom transcatheter closure was attempted. Diagnosis and follow-up were based on transesophageal echocardiography in the majority of cases. In all patients the "Sideris buttoned device" was used. A total of 19 patients (13 women) were treated for symptomatic ASD of the secundum type. The mean defect size before the intervention was 17 mm by echocardiography and 24 mm measured by balloon in the catheterization laboratory (stretched diameter) with a reduction to 4 mm after treatment measured by echocardiography. A total of 16 patients (5 women) underwent transcatheter closure of a PFO; 14 patients had had cerebral embolic events and 2 peripheral. During follow-up of 104 patient months, only one patient had a recurrent embolic event. He had at that time no occluder protecting the PFO after early embolization and extraction of the device. PMID- 8711464 TI - [Basis of radiation protection]. AB - After an introduction, three selected contributions to the 10th Course on Radiation Protection held at the University Hospital of Basel are presented. The principles of radiation protection and new Swiss legislation are discussed as the basis for radiological protection. Ways are proposed of reducing radiation exposure while optimizing the X-ray picture with a minimum dose to patient and personnel. Radiation effects from low doses. From the beginning, life on this planet has been exposed to ionizing radiation from natural sources. For about one century additional irradiation has reached us from man-made sources as well. In Switzerland the overall annual radiation exposure from ambient and man-made sources amounts to about 4 mSv. The terrestrial and cosmic radiation and natural radionuclids in the body cause about 1.17 mSv (29%). As much as 1.6 mSv (40%) results from exposure to radon and its progenies, primarily inside homes. Medical applications contribute approximately 1 mSv (26%) to the annual radiation exposure and releases from atomic weapons, nuclear facilities and miscellaneous industrial operations yield less than 0.12 mSv (< 5%) to the annual dose. Observations of detrimental radiation effects from intermediate to high doses are challenged by observations of biopositive adaptive responses and hormesis following low dose exposure. The important question, whether cellular adaptive response or hormesis could cause beneficial effects to the human organism that would outweigh the detrimental effects attributed to low radiation doses, remains to be resolved. Whether radiation exerts a detrimental, inhibitory, modifying or even beneficial effect is likely to result from identical molecular lesions but to depend upon their quantity, localization and time scale of initiation, as well as the specific responsiveness of the cellular systems involved. For matters of radiation protection the bionegative radiation effects are classified as deterministic effects or stochastic effects respectively. The various histopathological reactions of tissues and organs following localized tissue irradiation, and the radiation syndromes following total body irradiation, constitute the deterministic effects. There will be a threshold below which deterministic effects do not appear and spontaneous incidences are not known. For low dose risk considerations deterministic effects are of no significance. Genetic effects and carcinogenesis are said to be stochastic effects. Characteristically the probability of stochastic effects increases with dose but the severity of the effects is independent of the dose. The shape of the dose response relationship at intermediate to high dose levels is linear-quadratic. For exposure to low doses the response becomes linear, as is to be expected for a linear-quadratic function at low dose. No threshold is assumed for stochastic effects. The estimate of probability of fatal cancer by the ICRP is 4 x 10(-2) per Sv for the working population and 5 x 10(-2) per Sv for the total population. Their estimate of probability of serious hereditary disorders within the first two generations is 1 x 10(-2) per Sv. The highest probability coefficient is attributed to mental retardation following exposure in utero. Within the sensitive period at 8-15 weeks of gestation, a risk probability of 40 x 10(-2) per Sv is assumed but a threshold at 0.1 Sv is not excluded. Conclusions drawn from experiments, clinical observations and epidemiological studies following intermediate to high radiation exposures attribute a mutagenic and carcinogenic competence to all radiation doses. Microdosimetric considerations support this assumption. This conclusion cannot be confirmed experimentally nor by epidemiological studies of populations living under different conditions from natural sources of radiation. Nevertheless, a change in the present restrictive radiation protection policy does not yet appear appropriate. PMID- 8711465 TI - Uncertainty in estimated half-lives of PCBS in humans: impact on exposure assessment. AB - Estimates of the half-lives of PCBs in humans derived from successive body burden measurements are reviewed and found to vary widely whether based on congener specific or aggregate data. Variability due to differences in physiological processes among individuals and in congener properties is to be expected, but does not appear to be a complete explanation. Very short half-lives (i.e. < 1 year) are unlikely for those congeners most frequently found in human blood because the exposures required to sustain observed body burdens are too large to be easily explained. Very long half-lives ( > 10 years) may be artifacts of confounding by ongoing exposures (a common effect at low body burdens) and are also suspect. The loss of significant quantities of PCBs from the blood of occupationally exposed persons with half-lives of 2-6 years is comparatively well documented (i.e. has been observed in studies with relatively large numbers of subjects and high initial body burdens). Therefore, very long half-lives must be limited to subsets of congeners or of populations if they occur at all. The impact of the use of fixed estimates of half-lives drawn from the tails of the observed range on the evaluation of regulatory standards is shown to be substantial. PMID- 8711463 TI - [Tuberculous coxitis: diagnostic problems]. AB - Tuberculosis is still a significant disease in Switzerland, mainly because of the number of immigrants from endemic countries and the AIDS epidemic. The osteoarticular system is affected in a minority (3.1%) of cases; diagnosis is difficult but extremely important since classical treatment by medication started without delay brings about complete recovery, whereas lack of therapy can result in persistent pain and loss of joint function. Therefore, diagnostic procedures must not be limited to x-ray examination alone, but cultures and antibiotic sensitivity tests must be performed in order to select the appropriate antibiotic therapy. Cultures and histological analysis of the synovial membrane and fluid are the most reliable diagnostic tools. PMID- 8711467 TI - Gamma-ray activity and dose rate of brick samples from some areas of North West Frontier Province (NWFP), Pakistan. AB - It is important to measure natural radioactivity due to gamma-rays from building materials and consequently to determine the dose rate from these materials. This helps to implement precautionary measures whenever the dose rate is found to be above the recommended limits. A knowledge of gamma radioactivity is required by the building construction association to adopt preventive measures to mitigate or minimize the harmful effects of ionizing radiation. The aim of this work was to measure the gamma activity due to 226Ra, 232Th and 40K in bricks from the Peshawar, Nowshera and Mardan districts of NWFP of Pakistan. The radiological doses to the occupants from the brick floor of a wooden house were calculated. The average gamma dose equivalent rate was calculated to be 0.14 mSv per year, which is well below the external gamma dose limit of 0.46 mSv per year as given in UNSCEAR [2]. PMID- 8711466 TI - Validation of a methodology for modelling PCDD and PCDF intake via the foodchain. AB - The US EPA foodchain exposure methodology has been assessed, linking background concentrations of polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) in the atmosphere with eventual intake of these chemicals by humans via the foodchain. The methodology is assessed against background environmental concentrations of PCDD/Fs and the background daily adult intake of PCDD/Fs in the UK diet comprising meats, vegetables, dairy produce, fish and miscellaneous food groups. The environmental fate and transport of PCDD/Fs is estimated for each of the seventeen, 2,3,7,8-positional PCDD/F isomers individually from the atmospheric burden in the atmosphere and into the environmental medium or food product of interest. The model predicts PCDD/F deposition rate and soil concentration to within +/-50% of the measured I-TEQ values. For grass, the model underpredicts PCDD/F concentrations but the lack of a reliable and coherent data set precludes further investigation as to the likely causes. The model performs well on food products, with the exception of potatoes (under-predicts by a factor of 20) and 'other' vegetables (underpredicts by a factor of 3). The total modelled PCDD/F intake via the diet is 61 pg I-TEQ day-1 as opposed to the measured intakes of 69 pg I-TEQ day-1 (excluding plant foods) and 81 pg day-1 (including data for plant foods measured in a previous survey, and not representative of contemporary exposures). The major contributor to total PCDD/F intake is via milk and milk products, accounting for about 40% of the daily dietary intake. 'Hidden' fats and oils account for 35% of the PCDD/F intake, followed by the ingestion meat and meat products (12%). As a group, vegetables do not appear to contribute significantly to the total adult background intake of PCDD/Fs via the diet. PMID- 8711468 TI - Fenvalerate (alpha-cyano-m-phenoxy-benzyl alpha-isopropyl-p-chlorophenyl acetate) residues in and on okra fruits. AB - Okra crop was sprayed with sumicidin 20E (Fenvalerate 20% EC) at the rate of 300 ml/ha (C1), 375 ml/ha (C2) and 500 ml/ha (C3). Residues in fruit samples were determined by gas chromatography (Hewlett Packard 5890 A) at different time intervals to establish the safe waiting period for Fenvalerate. The initial deposits of Fenvalerate on okra fruits for C1, C2 and C3 were 1.4 ppm, 2.8 ppm and 4.5 ppm, respectively. Seven days after spraying, the initial deposits of C1, C2 and C3 decreased to 0.03 ppm, 0.05 ppm and 0.67 ppm, respectively. The residue values of Fenvalerate on okra fruits 7 days after spraying were below the tolerance limit (0.2 ppm; Anonymous, 1978; FAO, 1980) for C1 and C2, but above it in the case of C3. Ten days after spraying, the residue values of C1, C2 and C3 were below the tolerance limit. PMID- 8711469 TI - Defining misconduct. PMID- 8711471 TI - Preventing AIDS. PMID- 8711470 TI - AIDS politics. PMID- 8711473 TI - Funding crisis grips genome research. PMID- 8711472 TI - Ancient life on Mars? PMID- 8711474 TI - Learning defect identified in brain. PMID- 8711475 TI - Looking for clues to the mystery of life on earth. PMID- 8711476 TI - How congressional pressure shaped the 'Baltimore case'. PMID- 8711477 TI - A new receptor for growth hormone-release peptide. PMID- 8711479 TI - Salk: under new management. PMID- 8711478 TI - High-affinity potassium uptake in plants. PMID- 8711480 TI - Promega wins round in fight over Taq. PMID- 8711481 TI - More monkey business alleged at NYU. PMID- 8711482 TI - Life's last domain. PMID- 8711483 TI - An excellent lightness. PMID- 8711484 TI - Small nucleolar RNAs guide ribosomal RNA methylation. PMID- 8711485 TI - Possibilities of brain protection with tirilazad after cardiac arrest. AB - Cardiac arrest and resuscitation often create a cerebral insult caused by the initial cessation of blood flow, followed by the incomplete ischemia of cardiopulmonary resuscitation (low flow), and, following the return of spontaneous circulation, by the post-resuscitation syndrome. A cascade of physiologic, vascular, and biochemical events is set in motion, including changes in neuropeptides, electrolytes such as calcium and magnesium, excitatory neurotransmitters such as glutamate and acetylcholine, lymphokines such as interleukin-1, and arachidonic acid metabolites such as prostaglandins and leukotrienes; and formation of oxygen free radicals and lactic acid. Oxygen free radical-induced lipid peroxidation appears to increase tissue injury during and after brain ischemia. The 21-aminosteroid U74006F (tirilazad mesylate) is a novel inhibitor of lipid membrane peroxidation induced by oxygen free radicals, which has been shown, in animal models of subarachnoid hemorrhage, central nervous system trauma, and cerebral ischemia, to limit the extent of secondary tissue damage, thus improving functional recovery. Since tirilazad appears to have little or no behavioral or physiologic side effects, it appears to be an ideal agent for widespread brain ischemia prophylaxis. Tirilazad mesylate studies in out-of-hospital cardiac arrest are currently being planned. PMID- 8711486 TI - Thrombolytic therapy in acute myocardial infarction. AB - Despite major advances made over the last decade, mortality following acute myocardial infarction (MI) is still high even for patients treated with the "front-loaded" recombinant tissue-type plasminogen activator (rt-PA) regimen: 30 day mortality is 6.3%, and it is associated with fatal cerebral hemorrhage in 1.5%. Further improvement of short- and long-term prognosis can be achieved if infusion of the thrombolytic agent starts early, if reperfusion occurs more rapidly, and if a persistent thrombolysis in myocardial infarction (TIMI) grade 3 flow of the infarct-related artery can be achieved. These prerequisites for optimal preservation of ventricular function and predictors of a favorable outcome. New treatment strategies, e.g., more accelerated dosage regimens of established thrombolytic agents, new combination therapies, and new thrombolytic agents, with or without addition of more specific and more potent new antithrombotic agents, may further improve the efficacy of thrombolysis in acute MI during the late 1990s. PMID- 8711487 TI - Pulmonary thromboembolism: epidemiology and techniques of nuclear medicine. AB - Pulmonary thromboembolism is a very common disease (about 500,000 events per year in the United States). The most important risk factors seem to be age, cancer, previous surgery, trauma, coagulation disorders, and immobilization. Analysis of epidemiologic studies shows that the incidence of pulmonary embolism is underestimated clinically. Combined ventilation/perfusion scintigraphy is an established method for the diagnosis of pulmonary embolism, but the specificity of the method still is not high enough. Diagnostic strategies and new techniques in nuclear medicine will probably be able to improve the quality of the management of patients with clinically suspected pulmonary embolism. PMID- 8711488 TI - Preclinical management of thromboembolic disorders. AB - Preclinical management faces considerable diagnostic and therapeutic limitations. Patient history, physical examination, and few technical means (electrocardiogram [ECG], blood glucose measurement) must suffice to arrive at a suspected or definite diagnosis. Emergency treatment of most medical conditions inside the hospital differs from the preclinical setting. Critically ill patients must be transported to the hospital in emergency units with standardized equipment. Diagnosis of acute myocardial infarction relies on patient history, present symptoms, and a 12-lead ECG, and can be made with relative certainty. Therapeutic management focuses on reduction of myocardial oxygen consumption and optimizing oxygen availability. Monitoring for possible arrhythmias is essential. Few exceptional situations justify preclinical thrombolytic therapy. After a neurologic deficit has been diagnosed, the management of patients with stroke includes support of vital functions and admission to a hospital where computed tomography or magnetic resonance imaging is available. Pulmonary thromboembolism is frequently misdiagnosed even in the hospital. Preclinical physical examination, patient history, ECG, and pulse oximetry allow a positive diagnosis only in massive embolization. Anticoagulation with heparin is essential. If cardiopulmonary resuscitation is necessary, preclinical thrombolysis may be an option. Although deep vein thrombosis is difficult to recognize, classical symptoms of acute arterial occlusion are rarely missed. Treatment consists of general measures and anticoagulation. PMID- 8711489 TI - Pulmonary embolism. II. Diagnosis and treatment. AB - Pulmonary embolism is an often underestimated, underdiagnosed, and undertreated disease. As symptoms and signs of pulmonary embolism are nonspecific, the diagnosis still remains a challenge to the attending physician. Diagnostic and therapeutic procedures depend on the clinical presentation of the patient. First we must suspect pulmonary embolism and consider its likelihood in the presence of a number of clinical signs and symptoms. Once pulmonary embolism is suspected, heparin should be administered. Additional basic support is mandatory if required. If the patient's hemodynamic situation is stable, available preferable noninvasive diagnostic options should be considered to confirm or rule out the diagnosis of venous thromboembolism before further administration of anticoagulant or thrombolytic agents. If the patient's status has deteriorated, bedside diagnostic techniques should be applied to reinforce the suspicion or establish the diagnosis. To restore pulmonary perfusion more rapidly than conventional anticoagulation is suspected to do, several dosing regimens of thrombolytic agents are proposed, with recent interest in short-term thrombolysis and bolus lysis with urokinase or recombinant tissue plasminogen activator. If thrombolysis fails or is contraindicated, catheter embolectomy or surgical embolectomy is indicated. The main therapy is prevention. In this article, clinical assessment, imaging techniques, and therapeutic options described in the published literature are discussed and clinical experiences of an emergency department with a noninvasive diagnostic approach are described. PMID- 8711490 TI - Acute ischemic stroke. AB - While case fatality rates in ischemic stroke tend to decline, the total number of strokes is expected to increase further in the future because of more people reaching a greater age. Acute ischemic stroke, usually caused by chronic arterial hypertension, in most cases is induced either by a sudden loss of perfusion pressure or by thromboembolism. Reduction of oxygen supply to brain tissue then leads to a cascade of biochemical reactions. Cell death finally occurs after massive Ca2+ influx into the cell and breakdown of the membranes. A rim of viable tissue called the penumbra often exists around a central necrotic core within the ischemic region. The tissue compartment may be brought back to function if perfusion is restored within a short time. Since some cytotoxic reactions within ischemic tissue are irreversible, current efforts in stroke therapy focus on measures to decrease cellular vulnerability. Restoring perfusion remains the first important therapeutic goal. A variety of compounds have been tested for cytoprotection, but none can yet be recommended for routine clinical use. General management of stroke patients in every case should be implemented for emergency assessment, since the first few hours after onset are crucial for the outcome. PMID- 8711491 TI - Thrombolytic therapy in acute lower limb ischemia. AB - Surgical revascularization as the initial therapy in acute lower limb ischemia (ALLI) is associated with a high cumulative mortality and amputation rate. Catheter-directed delivery of low-dose thrombolytic agents (intra-arterial thrombolysis, IAT) offers the possibility for a gentle revascularization with a minimum of stress for the patients. In two randomized studies, the primary rates of revascularization, amputation, and mortality did not differ significantly between IAT and surgical revascularization. However, in one study the 6-month event-free survival rate was 85% in the IAT group, and 63% in the surgical group. Also in the second study the 12-month results were significantly better in the IAT group (event-free survival 75%) than in the surgical group (event free survival 52%). The high-dose urokinase regimen recommended by some authors in IAT is associated with an unacceptable cerebral bleeding rate of up to 2%. Low-dose recombinant tissue-type plasminogen activator (rt-PA) (0.02 to 0.05 mg/h) is the most suitable agent in IAT because of rapid lysis and low bleeding complications. Patients with ALLI, classified as viable or threatened without neurologic deficit, benefit most from the IAT as the initial therapy in ALLI. When IAT is performed as the initial therapy in ALLI, surgical intervention becomes unnecessary in approximately one-third of the patients. In another third the subsequent correction of the cause of the ALLI can be performed electively, which reduces mortality and morbidity rates. PMID- 8711492 TI - Disseminated intravascular coagulation: objective clinical and laboratory diagnosis, treatment, and assessment of therapeutic response. AB - Current concepts of the etiology, pathophysiology, clinical and laboratory diagnosis and management of fulminant and low-grade disseminated intravascular coagulation (DIC) have been presented. Considerable attention has been devoted to interrelationships within the hemostasis system. Only by clearly understanding the pathophysiological interrelationships can the clinician and laboratory scientist appreciate the divergent and wide spectrum of often confusing clinical and laboratory findings in patients with DIC. Objective clinical and laboratory criteria for diagnosis of DIC have been delineated, thus avoiding needless confusion and empirical decisions regarding the diagnosis. Many therapeutic decisions to be made are controversial and will remain so until more is published about specific therapeutic modalities and survival patterns. Also, therapy must be highly individualized depending on the nature of DIC, age, etiology of DIC, site and severity of hemorrhage or thrombosis, and hemodynamic and other clinical parameters. Also presented are clear criteria for severity of DIC and objective criteria for defining a response to therapy. Also, since it is often difficult for the individual physician to decide when to stop expensive therapy, objective criteria by which therapy may be stopped when continuation is likely fruitless are presented as a guideline. PMID- 8711493 TI - Blood rheology in emergency medicine. AB - Emergency conditions are generally associated with changes in blood rheology due to a systemic "acute-phase" reaction occurring within hours of the acute event. The acute-phase reaction includes changes in plasma protein patterns as well as in red cell properties. The deterioration of blood flow properties thus resulting is usually moderate and reversible. On the other hand, there are emergency situations caused or at least seriously aggravated by severe changes in blood rheology: circulatory shock, hyperviscosity syndromes, sickle cell crisis, and preeclampsia. In these conditions, special rheological therapeutic measures have to be taken into consideration. PMID- 8711494 TI - Management of acquired coagulation disorders in emergency and intensive-care medicine. AB - Coagulation disorders usually confront the emergency physician as bleeding episodes or as abnormalities of laboratory tests. Bleeding has to be treated aggressively, while pathological coagulation tests should be related to a more differentiated diagnosis at first. The most common causes of acquired coagulation disorders are liver disease, vitamin K deficiency, and disseminated intravascular coagulation (DIC). More rarely, inhibitors, external factors such as drugs or extracorporeal circulation, or other diseases such as amyloidosis are present. Since localized hemorrhage is the most common bleeding source in liver disease, endoscopic and surgical therapeutic measures, respectively, are warranted. Careful and balanced substitution therapy according to laboratory findings should be initiated simultaneously and should consist of fresh frozen plasma (FFP), which contains all components of the coagulation system physiologically balanced. Prothrombin complex concentrates should be used in emergency situations only, keeping their potential hazards in mind. Adequate vitamin K substitution is indicated in liver disease as well as in coagulopathy due to vitamin K deficiency. Management of DIC primarily consists of aggressive treatment of the underlying disease. Substitution therapy is difficult and should be carefully monitored by the adequate laboratory tests. FFP is the adequate source of both procoagulants and inhibitors but may cause certain problems. Heparin therapy can be beneficial but is not recommended generally. Antithrombin III substitution cannot be assumed as established therapy so far. Inhibitors can lead to bleeding, but the most common inhibitor, lupus anticoagulant, rather predisposes to thrombosis. In bleeding patients with inhibitors against single clotting factors, treatment consists of adequate substitution before initiating the diagnostic workup. PMID- 8711495 TI - Non-small cell lung cancer: meta-analysis of efficacy of chemotherapy. AB - The recent meta-analysis of 52 trials evaluating outcomes of non-small cell lung cancer patients treated with various modalities is discussed. From these data, three priorities can be proposed: (1) lung cancer physicians should alter their practices to include cisplatin-containing combination chemotherapy in their management protocols, (2) many more patients with lung cancer should be entered into clinical trials, and (3) the reporting of trial data, both response and toxicity data and quality of life assessments, needs to be improved. In addition, the poor outcome of non-small cell lung cancer patients, even those with the smallest volume of disease at the time of diagnosis, emphasizes the need for application of more aggressive treatments at the earliest possible stage of disease and the need for continued new drug development. PMID- 8711496 TI - Ifosfamide-based chemotherapy for non-small cell lung cancer: phase I/II studies at the University of Chicago. AB - Current clinical investigations in the palliative care setting for patients with non-small cell lung cancer are focused on new drugs and combinations. The goal of these studies is to identify more effective and/or less toxic therapy. At the University of Chicago, we have conducted phase I and II studies to integrate new single agents into novel combination chemotherapy regimens. Two such studies have combined the use of ifosfamide with either vinorelbine or paclitaxel. All these drugs have established single-agent activity in non-small cell lung cancer and a favorable toxicity spectrum. We describe the study rationale and design, and the preliminary data of these trials. PMID- 8711497 TI - The role of ifosfamide in the treatment of testicular and urothelial malignancies. AB - The activity of ifosfamide in genitourinary malignancies has been documented in testicular cancer and bladder cancer. The use of ifosfamide in germ cell tumors spans 20 years and has involved three distinct clinical settings: as a component of salvage therapy in cisplatin-resistant or recurrent disease, as part of initial therapy for patients with poor-risk disease, and as part of an ablative regimen for patients undergoing high-dose therapy with stem cell support. The use of ifosfamide in salvage therapy is well established, with approximately 40% to 50% of patients treated with a second-line ifosfamide-based regimen expected to achieve a complete response; however, only 25% of these patients will be long term disease-free survivors. The drug's incorporation into standard-dose first line regimens in patients with poor-risk disease has failed to improve the efficacy of therapy over standard, less toxic regimens. The value of ifosfamide as a component of high-dose salvage therapy with stem cell support in patients with refractory disease or of its increasing use as first-line therapy in patients with poor-risk features remains to be demonstrated. In urothelial carcinoma, data on the activity of ifosfamide are more sparse. Older trials in previously untreated patients in Japan and Egypt suggest an overall response rate of 30% to 40%, while a recent Eastern Cooperative Oncology Group trial in patients with prior chemotherapy reported a response rate of 20%. Ifosfamide has therefore been identified as one of six new active agents in urothelial cancer, and trials of combination regimens including ifosfamide are under way. The vinblastine/ifosfamide/gallium nitrate (VIG) combination was tested in a pilot study at Indiana University with a 67% overall response rate and in a confirmatory Eastern Cooperative Oncology Group phase II trial with a 56% response rate. Trials of ifosfamide plus paclitaxel with or without cisplatin are ongoing, but the role of ifosfamide in the routine therapy of urothelial malignancies remains to be determined. PMID- 8711498 TI - Ifosfamide in the treatment of lymphoma. AB - Currently, approximately 50% of patients with non-Hodgkin's lymphoma are eventually cured. Most patients who are cured reach a complete response after first-line chemotherapy, but a meaningful number of patients with partial response and those whose treatment fails or whose disease relapses can achieve a durable disease-free remission with effective salvage therapy. This usually involves multidrug intensive chemotherapy with or without bone marrow transplantation. Ifosfamide, an active drug in this setting, is a component of many of these salvage regimens. Some results with the combination of ifosfamide/etoposide, sometimes combined with mitoxantrone and/or cytarabine, are presented. Many relapsing lymphoma patients will require additional treatment after salvage chemotherapy, such as high-dose therapy with bone marrow transplantation, to realize a better long-term outcome. PMID- 8711499 TI - Epirubicin and ifosfamide in metastatic breast cancer. AB - In a randomized, phase II trial, we evaluated the effectiveness of continued chemotherapy with epirubicin/ ifosfamide versus unmaintained treatment interruption in advanced metastatic breast cancer. Three hundred fifty-seven patients were enrolled and 331 were evaluable for response. Complete response was achieved in 25 patients (8%) and partial response in 121 patients (37%). Pretreatment status correlated significantly with response (complete and partial response). While 54% of unpretreated patients responded, only 42% of the patients responded who had been pretreated with adjuvant chemotherapy and 33% who had been pretreated in the metastatic stage of disease; 69 patients (21%) had disease progression. Of 11 patients pretreated in both the adjuvant and metastatic setting, only two responded. Toxicity of treatment was mild, with leukopenia being the treatment-limiting factor. Thrombocyte levels were not altered significantly by treatment. Thus, there seems to be room for dose escalation using granulocyte colony-stimulating factor. There was no considerable cardiotoxicity, central nervous system toxicity, or cystitis observed. The low rate of cardiotoxicity appeared to be related to dose fractionation of epirubicin. After randomization of patients to treatment interruption versus continuation of chemotherapy, a longer relapse-free survival was observed for patients who continued chemotherapy (mean relapse-free survival, 2+ months); however, this did not translate into prolonged survival. The cumulative scores of toxicity and quality of life parameters showed increasing superiority for treatment interruption. Therefore, a strategy of treatment until maximum response and subsequent treatment interruption seems to be superior to treatment continuation. PMID- 8711500 TI - High-dose ifosfamide in the treatment of advanced soft tissue sarcomas. AB - Ifosfamide, an oxazaphosphorine analogue, is now commonly used in first-line anthracycline-containing regimens for advanced soft tissue sarcomas, based on its demonstrated single-agent activity in previously treated patients. At the Institut Gustave-Roussy, we performed a phase II study of high-dose ifosfamide without hematopoietic support in patients with adult advanced soft tissue sarcomas previously treated with a doxorubicin-based regimen. Our goal was to confirm a dose-response relationship for ifosfamide in soft tissue sarcoma. Ifosfamide was given at a dose of 4 g/m2/d over 3 days by continuous infusion with an equivalent dose of mesna; an additional 4 g/m2 of mesna was given on day 4. Among 36 evaluable patients, we observed no complete responses and 12 partial responses, for an overall response rate of 33%. No patients with leiomyosarcoma responded to treatment, and some histologic subtypes clearly showed better chemosensitivity than others. The major dose-limiting toxicities were hematologic. Our results confirm a dose-response effect for high-dose ifosfamide in soft tissue sarcomas, with a threshold of approximately 10 g/m2. PMID- 8711501 TI - The role of ifosfamide in small cell lung cancer. AB - The emergence of chemotherapy has dramatically improved both quality of life and survival in patients with small cell lung cancer (SCLC). Nonetheless, achieving long-term survival in SCLC patients has been a problem. Early studies of combination chemotherapy with cyclophosphamide/doxorubicin/vincristine (CAV) and cyclophosphamide/methotrexate/lomustine/vincristine (CMCV) reported impressive response rates in patients with SCLC. Ifosfamide, an analogue of cyclophosphamide, has demonstrated single-agent activity against SCLC, achieving overall response rates ranging from 5.6% to 76.5%. Because of this, and because of the agent's proven synergism in combination chemotherapy for recurrent testicular cancer and its relative non-myelosuppressive qualities (compared with cyclophosphamide), the incorporation of ifosfamide into combination chemotherapy for SCLC was rational. The Hoosier Oncology Group reported high response rates with VIP (ifosfamide combined with etoposide/cisplatin) in SCLC patients and proved the superiority of VIP over etoposide/cisplatin in patients with extensive disease. Presently, this group is evaluating the role of chronic oral etoposide as maintenance chemotherapy for patients with extensive SCLC that responds to initial VIP treatment. Salvage treatment with daily oral etoposide has also produced encouraging results, leading the Hoosier Oncology Group to incorporate oral etoposide as part of the VIP regimen (VoIP). It is currently unclear whether combination chemotherapy containing daily oral etoposide will have a major impact on survival. Further trials of combination chemotherapy with newer active agents like paclitaxel and topotecan, as well as with proven single agents like ifosfamide, are clearly warranted to improve the outcome of patients with SCLC. PMID- 8711502 TI - Ifosfamide in the treatment of pediatric malignancies. AB - Neuroblastomas, nephroblastomas, malignant mesenchymal tumors, including rhabdomyosarcomas, Ewing's sarcomas, osteosarcomas, brain tumors, and non Hodgkin's lymphomas respond to ifosfamide monotherapy. Ifosfamide has found an established place in the treatment of these pediatric malignancies. Ifosfamide has been shown to be an especially valuable agent in brain tumors in patients with a poor-prognosis medulloblastoma. The combination of ifosfamide with other cytotoxic drugs is more effective than ifosfamide medication alone. Ifosfamide has to be given with mesna to prevent bladder toxicity, but other toxic side effects, such as neurotoxicity, nephrotoxicity, and gonadal damage, are more difficult to prevent. Ifosfamide has proven to be an important asset in the treatment of cancer in children. PMID- 8711503 TI - [The central basis of discrete sampling behavior in the human pupillary control system]. AB - The human pupillary control system was investigated with double-pulse light dichoptic stimulation to explore its discrete sampling behavior. In this dichoptic stimulating method, the first pulse of the double pulse light stimulated one eye, of which the pupil area was recorded, and the second pulse stimulated the other eye (the contralateral eye). The experiment data demonstrated that the pupillary control system requires a minimum time interval, approximately 0.6 s, to elicit next transient response to the second light pulse, even though the two pulses were presented respectively to different eyes of the subject. The result supports the suggestion that the discrete sampling process exists in the pupillary control system. Moreover, it indicates that the minimum time interval is not caused by the retina adaptation, but determined by the central nerve system. PMID- 8711504 TI - [Inhibitory effects of hypothalamic arcuate nucleus or anterior pituitary stimulation on the nociceptive responses of thalamic parafascicular neurons]. AB - The effects of hypothalamic arcuate nucleus (ARC) or anterior lobe of pituitary (AL) stimulation on the nociceptive responses of thalamic parafascicular (Pf) neurons were studied in rats with electrophysiological technique. The results showed that ARC stimulation could inhibit the nociceptive discharges of Pf neurons, namely, the immediate inhibition because of its very short latency and duration. AL stimulation could also inhibit the nociceptive discharges of Pf neurons, but this inhibition was a delayed one because of its longer latency and duration. Hypophysectomy diminished the immediate inhibition due to ARC stimulation, while ARC lesion diminished the delayed inhibition due to AL stimulation. Both kinds of inhibition were blocked by dexamethasone pretreatment. The above results suggest that neuroendocrine relationship between ARC and AL is involved in pain modulation. PMID- 8711505 TI - [Effect of (-) stepholidine on dopamine turnover in various brain regions]. AB - It has been shown before that DA antagonist (-) stepholidine [(-) SPD] changes the function of feedback regulation in the striatum. In order to compare the effect of the drug on other brain DA systems, the DA content and DOPAC level in brain areas or nuclei of rat were measured by HPLC-ECD. It was found that (-) SPD (10 mg/kg, ip) did not significantly affect the DA contents in the striatum, olfactory bulb, N. accumbens, substantia nigra, ventral tegmentum area, N. amygdala, hypothalamus, pituitary, piriform cortex, but did markedly increase the DOPAC levels in these brain areas or nuclei, and thus increase the DA turnover (DOPAC/DA). However, (-) SPD (10 mg/kg, ip) did not significantly affect either the DA contents or DOPAC levels in the prefrontal cortex and cingulate cortex, thus leaving the DA turnover in these areas unchanges. These results indicated that (-) SPD did affect the nigro-striatal, midbrain-limbic and hypothalamus pituitary DA systems, but not the midbrain-cortex DA system. It is supposed that this difference may be due to the modulation by presynaptic DA autoreceptors in the major brain areas of DA nerve endings or neurosoma, but none in the cortex DA nerve endings. PMID- 8711506 TI - [Effect of acute hypoxia on L-type calcium current of the ventricular myocytes of guinea-pig]. AB - When the conventional whole cell recording technique is used to study the inward calcium current in cardiac cells, the "Run down" phenomenon of calcium channel would prevent sufficient time of recording desirable for adequate experimental analysis. The "Run down" phenomenon could be minimized by using nystatin-whole cell recording technique in isolated guinea-pig ventricular cells. The inward L type calcium current could be maintained at a steady level for up to more than 100 min, showing the presence of an endogenous steady calcium ion buffering mechanism. With nystatin-whole cell recording, the L-type calcium current after 10 min acute hypoxia (Po2 4 +/- 0.7 kPa) was inhibited (peak amplitude decreased) and the I-V relation was shifted upward. The inward calcium current showed no recovery after 10 min reoxygenation. The peak amplitude was lower than that of the control. The results suggested that the decrease of action potential duration (APD) under acute hypoxia was not only due to increase of outward potassium current, but also a decrease of inward calcium current. All these phenomena may be related to some inhibition of phosphorylation of the L-type calcium channel in the cardiac cells under hypoxia. PMID- 8711507 TI - [Whole cell recording of sodium, potassium and calcium currents of cultured neonatal rat sympathetic neurons]. AB - Na, K and Ca currents and other electrophysiological characteristics of cultured neonatal rat superior cervical sympathetic neurons were studied using whole cell clamp technique. The mean passive and active membrane properties measured are as follows: resting membrane potential, -51 +/- 6 mV; input resistance, 1432 +/- 389 M omega; time constant, 130 +/- 32 ms; amplitude of action potential, 96 +/- 10 mV; overshoot, 42 +/- 6 mV. Na, K and Ca currents were isolated upon pharmacological manipulations. The predominant type of K current was a noninactivating delayed rectifier. Voltage-clamp studies also showed the presence of a high voltage-activated sustained inward Ca current, while low voltage could not elicit any transient Ca current. PMID- 8711508 TI - [Hypoxia stimulates the expression of HSP70 in pulmonary artery smooth muscle cells]. AB - Hypoxic pulmonary hypertension is characterized by remodeling of pulmonary artery, proliferation and migration of pulmonary artery smooth muscle cells (PASMC), the mechanism of which was studied by observing the effect of hypoxia on DNA synthesis, cell cycle phase and transcription of heat shock protein 70 (HSP70). Our results indicate that hypoxia increases DNA synthesis, enhances the mitogenic effect of ET-1 on PASMC, stimulates quiescent PASMC entering S phase from G0/G1 phase and up-regulates the mRNA level of HSP70. Heat shock proteins are conservative and important stress proteins rendering the cells more tolerable to dangerous stresses by promoting growth and survival of the cells. All these findings suggest that heat shock proteins might also be important to the proliferation of PASMC induced by hypoxia and relevant to the study of the mechanism of hypoxic vascular remodeling. PMID- 8711510 TI - [Relative changes between calcium content and number of atrial specific granules induced by water and salt loading]. AB - Rats prepared by either deprivation of water intake for 5 d or drinking 2% NaCl solution for 4 d were sacrificed for dissection of auricle, and making ultrathin sections. The numerical density (Nv) of atrial specific granules (ASG) were counted by using electron microscopic morphometry, and Ca concentration in ASG and in terminal cisterna of sarcoplasmic reticulum (TSR) was determined by using electron microscopic X-ray microanalysis. In the drinking 2% NaCl solution group, the Nv of ASG decreased from 6.02 +/- 2.30 to 2.96 +/- 1.62/microns 3 (P < 0.01) and all companied by a decrease of Ca concentration from 64 +/- 16 to 38 +/- 21 mmol/kg (P < 0.01), while the secretion of atrial natriuretic peptide (ANP) showed some increase. Water deprivation induced all the contrary results. However, the Ca concentration in TSR was not affected by water-salt loading. It is suggested that when the atrial cardiocytes were stimulated by salt loading, Ca in ASG was released into cytosol and thus promoted fusion of ASG membrane with sarcolemma which resulted in release of ANP. Therefore, it is considered that ASG as an intracellular Ca store participates in the stimulus-secretion coupling of ANP through the release of Ca ions. PMID- 8711509 TI - [8%-9% and 12%-13% hypoxic gas induced free radicals generation in rat's left and right myocardium]. AB - Alpha-phenyl-tert-butylnitrone (PBN) was administered intravenously to capture free radicals of rat's myocardium. Rats were exposed to hypoxic gas (8%-9% O2 in N2) for 15 min. The ESR (electron spin resonance) signal intensity of PBN-spin adduct in the left myocardium increased significantly as compared with the normoxia group (n = 5, P < 0.05), but in the right myocardium there was no significant changes between hypoxia and normoxia. Rats exposed to hypoxic gas (12%-13% O2 in N2) were divided into four groups: I (hypoxia for 15 min), II (hypoxia for 60 min), III (hypoxia for 30 min/normoxia for 15 min/hypoxia for 30 min) and IV (injected MPEG- SOD intravenously before hypoxia for 60 min). The ESR signal intensity of PBN-spin adduct of left and right myocardium in group II increased significantly as compared with normoxia group (n = 5, P < 0.01), but the ESR signal intensity of group I didn't show obvious change as compared with normoxia group (n = 5, P > 0.05). In the right myocardium of group III the ESR signal intensity of PBN-spin adduct decreased significantly as compared with group II (n = 5, P < 0.05) and in the left myocardium did not decrease evidently. In the left myocardium of group IV the ESR signal intensity of PBN-spin adduct decreased evidently as compared with group II (n = 5, P < 0.05) and that in the right myocardium did not decrease evidently. When the rats were exposed to 8%-9% hypoxic gas for 15 min and 12%-13% hypoxic gas for 60 min, the SOD (superoxide dismutase, EC 1.11.1.9) activity of myocardium decreased and the content of MDA (malondialdehyde) increased significantly (n = 8, P < 0.05 or P < 0.01). The above results suggested that one way of myocardium free radical gereration may be relevant to decrease of SOD activity. The generation of free radicals pertained chiefly to superoxide free radical in the left myocardium and the membrane structure of myocardium cells might have been damaged largely during hypoxia. PMID- 8711511 TI - [Effect of electrical stimulation of afferent renal nerve on arterial blood pressure, heart rate and vasopressin in rabbits]. AB - The effect of electrical stimulation of afferent renal nerve (ARN) on cardiovascular response, the synthesis and release of vasopressin were studied in rabbits. During the course of the experiment, the pathway of ARN to central nerve system was also analyzed. The results showed that electrical stimulation of ARN elicited significantly decrease of mean arteral blood pressure and heart rate as well as inhibition of cervical sympathetic nerve activity. In the event of the above physiological changes, the AVP concentration in supraoptic nucleus (SON), paraventricular nucleus (PVN) and plasma was increased, but that in hypophysis was decreased. Injection of sodium nitropruside (SNP) or AVPa indicated that increase of AVP release was due directly to stimulation of ARN. Nodose ganglionectomy or transversal section of spinal cord at different levels suggested that the main afferent pathway of ARN to higher level of central nerve system entered into spinal cord at T5-L2. PMID- 8711512 TI - [Seasonal variation in the ultrastructure and calcium uptake rate of cardiac sarcoplasmic reticulum in ground sqirrel]. AB - The ultrastructure and calcium uptake rate of the cardiac sarcoplasmic reticulum (SR) was quantatively compared between the ground squirrels (Citellus dauricus) in hibernating and non-hibernating season. (1) In our ultrastructural investigation, OsFeCN post-fixation method was used to enhance the staining of SR, which made the SR structure more recognizable. (2) The results of morphometrical analysis demonstrated that the volume density of SR and lipid droplet in the cardio-myocyte was significantly higher (P < 0.01) in the hibernating animal than in the summer active one, and the subcompartment of SR with this difference was non-junctional SR. (3) The cardiac SR from hibernating ground squirrels had a higher (P < 0.01) calcium uptake rate as measured by the absorbence of arsenazo-III Ca2+ indicator, than that from autumn active ones. Altogether, our results suggested that the function of SR in the uptake of cytosolic calcium was enhanced in hibernating season, which may play an important role in the cold tolerance of myocardium of a hibernator. PMID- 8711513 TI - [The effect of GM-CSF/IL-3 on the grafting efficiency of CD34+ hemopoietic progenitor cells from umbilical cord blood]. AB - CD34+ cells were isolated from human umbilical cord blood by using a high gradient magnetic cell sorting system with anti-CD34 monoclonal antibody coated with microbeads, and then inoculated onto a pre-formed irradiated bone marrow stroma to evaluate the grafting efficiency of CD34+ cells with rhGM-CSF or IL-3, alone or in combination. The results showed that only 36% CD34+ cells in the control, and 68 - 89.6% in the groups with growth factors treatment seeded to the stroma. A short incubation of CD34+ cells with growth factors could rapidly reconstitute a long-term hemopoiesis in the long-term liquid culture system. This demonstrates that a brief treatment of CD34+ cells with GM-CSF or IL-3 can improve the grafting efficiency of transplanted umbilical cord blood cells. PMID- 8711514 TI - [Neural structural relations between the chemosensitive areas of the ventrolateral medulllary surface and the medullary nuclei in rats]. AB - Experiments were done on 30 chloralose-urethane anesthetized Sprague-Dawley strain rats. The ventrolateral medullary surface was exposed and a small millipore filter paper soaked with horseradish peroxidase (HRP) was applied locally to different parts of the underlying chemosensitive areas separately, trying to identify location of the HRP-stained cells. (1) After HRP application to the rostral area (n = 10), quite a number of labelled cells were distributed at the ventrolateral part of the nucleus of trapezoid body (Tz), the lateroventral periolivary nucleus (LVPO) and the retrotrapezoid nucleus (NRT). (2) After HRP application to the caudal area (n = 10), HRP labelled cells were mainly located at the lateral reticular nucleus (LRt) and the ventral part of at the lateral paragigantocellular nucleus LPGi. (3) After HRP application to the intermediate area (n = 6), only 8 HRP labelled cells were observed at the (LPGi) in 6 rats. (4) After HRP application to the control area (n = 4), only 6 HRP labelled cells were found at the rostral ventrolateral reticular nucleus (RVL). The above findings demonstrated the existence of the neural structural relations between the chemosensitive areas of the ventrolateral medullary surface and Tz, LVPO, NRT, LPGi, LRt, which supports that all these nuclei might participate in the central chemoreception of respiration. PMID- 8711515 TI - [Effects of alpha 1-adrenergic receptor subtypes activation on proliferation of smooth muscle cells in rabbit aorta]. AB - The effect of alpha 1-Adrenoceptor (alpha 1-AR) and its subtypes, alpha 1A and alpha 1B, activation on the proliferation of cultured vascular smooth muscle cells (VSMC) was studied by [3H]-thymidine (3H-TdR) incorporation and in situ hybridization. Norepinephrine (NE) promotes DNA synthesis of VSMC. The action was induced by activation of alpha 1A-AR subtype and increase of c-fos gene expression. However, the DNA synthesis was inhibited by activation of alpha 1B-AR subtype. There was a synergetic effect on VSMC DNA synthesis between NE and neuropeptide Y (NPY). But only additional effect was found between NE and Ang II. More over, it was found that alpha 1B-AR subtype showed significant constitutive inhibitory effect of VSMC DNA synthesis. PMID- 8711516 TI - [Effects of lesion or stimulation of pituitary gland and hypothalamic arcuate nucleus on pain modulation in rats]. AB - Relationship between pituitary gland and hypothalamic arcuate nucleus (ARC) in the modulation of pain threshold was investigated in Wistar rats with method of focal lesion and stimulation of pituitary gland. Experimental results indicated that electrolytic lesion of the pituitary intermediate and anterior lobes resulted in a decrease of pain threshold and disappearance of the analgesia immediately produced by ARC stimulation. Stimulation of the same area of pituitary gland could induce a delayed analgesia, which could be blocked by ARC lesion. PMID- 8711517 TI - [Acute hypoxia induce free radicals generation in rat's cortex and hippocampus]. AB - Anesthetized rats were ventilated artificially to 10.5%-11.0% O2 in N2 (equivalent to altitude about 5 000 m). PBN (alpha-phenyl-ter-butylnitrone) was injected intravenously 5 min before hypoxia. The experiment results showed that the ESR (electron spin resonance) signal intensity of PBN-spin adduct in cortex and hippocampus both increased significantly during hypoxia (15 min) as compared with normoxia group (n = 5, P < 0.01). In the third group of rats injected with MPEG-SOD (monomethoxy-polyethylene glycol-superoxide dismutase) intravenously 20 min before hypoxia, the ESR signal intensity of PBN-spin adduct in cortex and hippocampus did not decrease significantly under hypoxia for 15 min (compared with hypoxia group, n = 5, P > 0.05). This may be due to the fact that MPEG-SOD could not pass the blood-brain barrier owing to its large molecular weight. The enzyme activity of SOD (superoxide dismutase. EC 1.15.1.1), GSH-Px (Glutathione peroxidase, EC 1.11.1.9) and content of MDA (malondialdehyde) was also determined. The results showed that the hypoxia had no effect on the SOD, GSH-Px activity and MDA content (n = 7 or n = 8, P > 0.05). The above results suggested that ventilation to 10.5%-11.0% O2 in N2 15 min induced free radical generation in rat's cortex and hippocampus, but this generation was not due to decrease of SOD, GSH-Px activity in cortex and hippocampus. PMID- 8711519 TI - [Concerning geriatric care]. PMID- 8711518 TI - [Intracerebroventricular injection of interleukin 1 receptor antagonist antagonises on the generation of immuno-suppressive factor in restraint mice]. AB - Our previous work showed that a lymphocyte proliferation suppressing factor could be found in the serum of restraint mice. In the present work, it was found that intracerebroventricular (icv) injection of IL-1 receptor antagonist (IL-1Ra) was found capable of suppressing the production of such a serum protein under restraint stress. Nearly complete suppression could be achieved by 5.0 micrograms IL-1Ra. Intracerebroventricular injection of IL-1 beta (1 pg), however, increased the generation of the supressive protein. Neither intraperitoneal (ip) injection of IL-1Ra or IL-1 beta had any effect on the generation of the protein. The fact that icv. injection of a very small dose of IL-1 beta (0.06 fmol) was effective on the generation of the supressive protein led us to suggest that IL-1 in brain might act as an important mediator between CNS and the immune system. PMID- 8711520 TI - [When is it our desire? The family history]. PMID- 8711521 TI - [A course in psychiatry]. PMID- 8711522 TI - [A unique diploma, a unique profession...]. PMID- 8711523 TI - [Drug abuse and health district: value and role of an interface]. PMID- 8711525 TI - [Twenty years of nursing school at the Charles Perrens Hospital Center in Bordeaux]. PMID- 8711524 TI - [Professional autonomy of psychiatric nurses]. PMID- 8711526 TI - [The bad son]. PMID- 8711527 TI - [Foundations of psychiatric nursing: history]. PMID- 8711528 TI - [Why have educational requirements?]. PMID- 8711529 TI - [Diagnosis of perianal and perirectal fistulas and abscesses]. PMID- 8711530 TI - [Symptomatic endometriosis of the large intestine--a case report]. PMID- 8711531 TI - [Superior mesenteric artery syndrome]. PMID- 8711532 TI - [Malignant schwannoma of the abdomen]. PMID- 8711533 TI - [Lupus anticoagulants in patients with systemic lupus erythematosus]. AB - Lupus anticoagulants (LA) and anticardiolipin antibodies (aCL) are known as thrombosis-related antiphospholipid antibodies. LA is not as well characterized as aCL, and the relation between LA and aCL is not clarified. Since standardized method for the detection of LA has not been established, we measured LA activities in outpatients with SLE by using two different methods (KCT and dRVVT), and analyzed the characteristics of LA in SLE. LA was detected in 29.8% of all samples (14.3% in both methods, 15.5% in one method). IgG-aCL and IgM-aCL was detected in 38% and 20%, respectively, of all LA positive samples. Though a good correlation was observed between LA activities and IgG-aCL levels, a considerable number of LA positive samples were negative for aCL. This indicated the presence of factors with LA activity other than aCL. On the contrary there was also a high percentage of LA negative samples with positive aCL (42.4% in IgG aCL, 47.4% in IgM-aCL), suggesting the presence of aCL with poor or low LA activity. These findings showed the heterogeneity of antiphospholipid antibodies both in LA and in aCL. The platelet function tests showed increased platelet adhesiveness and normal platelet aggregation in LA positive patients with SLE even in the inactive phase. The serum levels of factors such as protein C, protein S, antithrombin III and thrombomodulin were within normal range. Clinical features such as hemolytic anemia, thrombosis and abortion were more frequently observed in LA positive population than in LA negative population. The clinical features tend to be different between patients with dRVVT-LA and those with KCT LA, though not significant. Because of the heterogeneity in LA, a combination of more than two different methods including dRVVT was recommended for the detection and the evaluation of LA. PMID- 8711534 TI - [Relationship between higher cortical dysfunction and the findings of magnetic resonance imaging in systemic lupus erythematosus]. AB - The relationship between systemic lupus erythematosus (SLE) and organic lesions was investigated by magnetic resonance imaging (MRI) to clarify the etiology of higher cortical dysfunction in SLE. The subjects were 10 patients with SLE, and higher cortical dysfunction was observed in 8 (80%) of the 10 patients. 5 (82.5%) of the 8 patients showed abnormal MRI findings. The findings of higher cortical dysfunction were consistent with the MRI findings in 1 of the 5 patients, but not in the remaining four. MRI revealed no lesion despite the presence of higher cortical dysfunction in three patients. These results suggest that the association of organic changes and functional changes in cerebral nerve cells is important in etiology of higher cortical dysfunction in SLE. PMID- 8711535 TI - [Clinical characteristics and genetic background of secondary amyloidosis associated with rheumatoid arthritis in Japanese]. AB - In order to examine the clinical characteristics and genetic background of secondary amyloidosis associated with rheumatoid arthritis, we analyzed clinical features and HLA typing of 85 patients in a multicenter study. Eighty-five patients with secondary amyloidosis associated RA were studied. The diagnosis of secondary amyloidosis were made on histological findings by biopsy or autopsy. The most common biopsy site was gastrointestinal tract (79.5%). Clinical symptom and the frequency at the time of diagnosis were; diarrhea (35 cases), abdominal pain (22 cases) and vomiting and nausea (16 cases). Abnormalities and the frequency in a laboratory test included proteinuria (49 cases), increased serum creatinine (32 cases), anemia (30 cases) and hematuria (15 cases). Twenty-eight patients were dead and 57 patients were alive at the time of the study. The average duration between diagnosis of amyloidosis and death was 19.4 +/- 18.5 (SD) months among the dead patients. The average duration after diagnosis of amyloidosis was 24.2 +/- 19.5 (SD) months in surviving patients. The causes of death were renal failure complicated with heart failure (6 patients), heart failure alone (3 patients) and renal failure alone (2 patients). Fifty-nine patients in the control group who were negative to amyloid deposition on biopsies at more than one site in the gastrointestinal tract, were clinically compared with patients in the amyloidosis group. No difference were noted in the age of RA occurrence and the stage between the two groups. As to the class, however, the number of patients with severe functional disorder (class 3 or severe) was larger in the amyloidosis group. There were no significant difference between the two groups in Lansbury's activity index. On hematology, biochemistry and urinalysis, the incidences of increased white blood cell count, anemia, increased platelet count, increased serum creatinine, hypoproteinemia, hypoalbuminemia, increased IgA, and increased urine and blood BMG were statistically significantly higher in the amyloidosis group than in the control group. HLA-A, -B, -C, and DR-locus antigens were compared in the 53 patients in the amyloidosis group and in the 59 subjects in the control group. There were no significant differences in frequency of HLA-A, and -B antigens between two groups. Frequency of CW7 antigen was significantly decreased in the amyloidosis group (13.2%) than in the control group (39.0%). Frequency of DR1 antigen was decreased in the amyloidosis group (3.8%) than in the control group (22.0%), although the difference was not significant. These findings suggest the possible involvement of genetic factors in the occurrence of amyloidosis. It is suggested that the occurrence of amyloidosis is suppressed by some genes which are linked with CW7 antigen. PMID- 8711537 TI - [A case of Sjogren's syndrome with an eyelid tumor, a so-termed pseudolymphoma of the iacrimal gland]. AB - The female subject, 64 years old, subjectively noticed a mild dryness of the eyes and bilateral, submandibular swelling in 1988. The clinical symptom was mild, and such autoantibody and hyper-gammaglobulinemia as often observed in Sjogren's syndrome were not recognized. However, by judging from the results of gum test (9 mL/10 min), salivary gland scan (Grade 2) and the labial biopsy, we diagnosed the case as Sjogren's syndrome. We then administered prednisolone at 40 mg at the onset of treatment and gradually decreasing the dosage over 3.5 years, and the symptoms improved. As an eyelid tumor in the left eye was noticed in 1991 and began to enlarge, the subject was hospitalized in June of 1992. The biopsy of the tumor (2.0 x 1.7 cm) showed marked polyclonal lymphoproliferation with lymphoid follicles which was determined by immunohistological staining. The case was a patient with a lymphoproliferative lesion from a lacrimal gland which is equal to a so-termed pseudolymphoma. Since there are some reports concerning the transition from pseudolymphoma to malignant lymphoma, this patient should be followed carefully. PMID- 8711536 TI - [Bucillamin induced lung injury in rheumatoid arthritis]. AB - Thirteen cases with rheumatoid arthritis who experienced lung injury during the treatment with bucillamine (Bc), about whom the questionnaires were answered by the physicians and whose X-ray films could be rechecked, were studied. Nine cases out of the 13 showed patchy mottled infiltrates in the bilateral center sparing the periphery, and the other 4 showed diffuse infiltrates. In these 9, serum gamma-globulin level decreased when lung injury appeared. The gamma-globulin level before the start of Bc administration, the level when lung injury appeared, the gamma-globulin decrease, and its ratio to the level before Bc were 1790 +/- 661 mg/dl, 1297 +/- 666 mg/dl, 459 +/- 320 mg/dl and 29.1 +/- 18.0%, respectively. In 5 out of the 9, gamma-globulin level reincreased when they recovered from the injury: 2 out of the 5 showed the reincrease even after steroid therapy. The data were obtained only from 2 out of the 4 with diffuse pattern in X-ray, and the decrease and the decrease ratio were 200 mg/dl (5.8%) in 1, and 49 mg/dl (3.6%) in the other. The characteristics of Bc-induced lung injury might be mottled infiltrates in the center appearing concurrently with serum immunoglobulin decrease. PMID- 8711538 TI - [Three cases with juvenile rheumatoid arthritis who showed hypocomplementemia related to disease activity]. AB - We encountered three cases of juvenile rheumatoid arthritis which showed a depressed serum complement level during the clinical course. Circulating immune complex and increased secondary fibrinolysis were detected in all cases. The laboratory data suggested the presence of vesculitis. Cutaneous vasculitis was observed by biopsy in 2 cases and abnormal respiratory function in one cases. These 3 cases were in contrast to other 95 JRA cases treated in our department who showed normal serum complement levels. Estimation of serum complement may, therefore be useful to predict the vascular complication and refractoriness to therapy in JRA. PMID- 8711539 TI - [A case of eosinophilic fasciitis with excessive increase of muscle enzyme]. AB - A sixty-year old woman had muscle pain of her extremities and stiffness in her fingers after unusual physical exertion. On admission she developed indurative swellings of the upper extremities and flexion contractures of her fingers. Laboratory data showed peripheral blood eosinophilia and excessive increase of muscle enzymes. Electromyography showed a myopathic pattern and magnetic resonance imaging showed findings of myositis. We performed a full thickness biopsy from skin to muscle. In histopathologic studies, collagenous hypertrophy of the fascia and perifascicular atrophy of muscle fibers and infiltrates on eosinophiles in the interstitial tissue were found. She was treated with prednisolone (60 mg/day). Clinical symptoms showed improvement and her peripheral blood eosinophiles and serum muscle enzymes deminished quickly. In this case, we suggest that the characteristic histopathologic findings in eosinophilic faciitis is not fascia alone, but includes muscle. We stress that eosinophilic faciitis and polymyositis are closely related. PMID- 8711540 TI - [Human gene therapy]. PMID- 8711541 TI - [Assessment of disease activity in systemic lupus erythematosus]. PMID- 8711542 TI - [Studies on anti-DNA antibodies associated with renal involvement by enzyme linked immunosorbent assay in patients with systemic lupus erythematosus]. AB - It has been reported that a significantly higher incidence of lupus nephritis was found in patients with high avidity anti-DNA antibodies. Radioimmunoassay (Farr's assay) is a method which enables to detect high avidity anti-DNA antibodies, whereas enzyme-linked immunosorbent assay (ELISA) can detect anti-DNA antibodies from low to high avidity. There are, however some patients who had high levels of anti-DNA antibodies by Farr's assay without renal involvement. In this study, ELISA was developed to detect IgG anti-DNA antibodies highly associated with lupus nephritis by changing salt concentration of reaction buffer solution. Levels of a fraction, we call [0.1 M - 0.3 M] fraction, which was obtained from the antibody levels measured under 0.1 molar of sodium chloride (NaCl) subtracted by antibodies levels under 0.3 molar of NaCl solution were found to be significantly higher in patients with urinary protein (p = 0.0074) and low serum complement (C 3 less than 50 mg/dl; p = 0.0026, C 4 less than 10 mg/dl; p = 0.0280 and CH 50 less than 30 U/mL; p = 0.0662). Among the patients with hypocomplementemia, levels of [0.1 M - 0.3 M] fraction were significantly higher in patients with urinary protein than in patients without renal involvement. This fraction might be consistent with anti-DNA antibodies with intermediate avidity that are related to lupus nephritis. The ELISA procedure established in this study is showed to be a available method to detect anti-DNA antibodies associated with renal disease in patients with systemic lupus erythematosus. PMID- 8711544 TI - Medical research--inferential errors. PMID- 8711543 TI - Pre-eclampsia and maternal mortality--a universal problem. PMID- 8711545 TI - South African medical research--ugly stepsister or fairy godmother? PMID- 8711546 TI - Can we support high-tech research in South Africa? PMID- 8711548 TI - Comparative costs of mobile and fixed-clinic primary health care services. AB - With restructuring and rationalisation of health services in South Africa imminent, the development of methods for comparing and evaluating health services is of great importance at both national and local level, including comparisons of cost-efficiency and cost-effectiveness. The costs of different methods of delivering primary health care in a local authority through mobile and fixed clinic services have been analysed and aspects of their cost-efficiency compared. The information gained from such an analysis can be used for management purposes to optimise both the use of resources and the quality of service provided at local level. PMID- 8711547 TI - Sexual behaviour, contraceptive practice and reproductive health among school adolescents in rural Transkei. AB - OBJECTIVES: To determine the patterns of sexual maturation, sexual behaviour, contraceptive practice and reproductive health among Transkeian adolescents. DESIGN: Cross-sectional descriptive study, using self administered questionnaires. SETTING: Twenty-six schools in 22 rural districts of the Transkei region of the Eastern Cape. SUBJECTS: Standard 5, 6 and 7 pupils of both sexes. OUTCOME MEASURES: Demographic and social characteristics, maturational and sexual behavioural milestones, and the prevalence of contraceptive use, pregnancies and sexually transmitted diseases (STDs). RESULTS: Data from 1,072 girls and 903 boys were analysed. The mean ages (+/-SD) of the girls and boys were 15.29 +/- 1.89 and 16.25 +/- 2.05 years, respectively. Menarche and semenarche occurred at 13.90 +/- 1.23 and 15.12 +/- 1.58 years in girls and boys, respectively. Overall, 76% of the girls and 90.1% of the boys in this survey were already sexually experienced. The age of initiation of sexual activity was positively correlated with the age of first dating and the age of menarche and semenarche. The boys initiated sexual activity at an earlier age (13.43 v. 14.86 years, P = 0.0000), had sexual intercourse more regularly (61.6% v. 42.3%, P = 0.0000) and more frequently, and had more lifetime sexual partners (mean of 3.27 v. 1.35, P = 0.0000) than the girls did. Nearly twice as many sexually experienced boys as girls had a history of STD (48% v. 25%, P = 0.0000) and, of these, 19.1% of the boys and 6.5% of the girls had a history of genital ulcer disease. The prevalence of whether contraceptives had ever been used was only 23% among sexually experienced girls and, of these, only 19.4% used condoms. Surprisingly, 62.1% of the sexually experienced boys had used condoms, and of these nearly one-third enjoyed using them. The prevalence of adolescent schoolgirl pregnancy was 31.3%. CONCLUSIONS: Sexual maturation occurs at an earlier age than previously among rural Transkeian adolescents. This is associated with early initiation and a high level of sexual activity, low contraceptive usage, and a high rate of adolescent pregnancy and STDs, which therefore expose adolescents to a high risk of HIV infection. PMID- 8711549 TI - Inadequate pre-season preparation of schoolboy rugby players--a survey of players at 25 Cape Province high schools. AB - OBJECTIVE: To establish the previous rugby experience, the knowledge and the use of injury prevention techniques by South African schoolboy rugby players. DESIGN: Before the first full-contact match of the 1991 rugby season, 2,330 players completed a detailed questionnaire. SETTING: High schools in the Cape Province. Three thousand three hundred and thirty players from 25 schools selected because of a record of excellence in and commitment to schoolboy rugby. MAIN OUTCOME MEASURES: Personal data including age, height, mass and rugby experience; history and nature of previous rugby injuries; knowledge of techniques known to prevent rugby injuries; parental and personal reasons for playing rugby. RESULTS: The incidence and distribution of the different types of injuries previously sustained by the players were the same as those identified prospectively in the same population. A-team players were significantly heavier and taller than players in lower teams in most playing positions in most age groups. Although A team players were more likely to participate in pre-season endurance or strength training, fewer than 40% of players overall trained adequately in the pre-season. Less than 30 minutes was allocated to the practice of tackling and falling techniques prior to the first full-contact match. Knowledge of different high risk situations during play was generally sketchy, and only 24% of players wore gumguards all the time. More fathers (84%) than mothers (63%) encouraged their sons to play rugby. CONCLUSIONS: The results indicate that the incidence and nature of the injuries reported retrospectively were similar to those reported in prospective studies at the same schools. The players' knowledge of techniques known to prevent rugby injuries was inadequate and too little attention was paid at the start of the rugby season to training and coaching techniques to reduce injury risk. Coaching errors may therefore have predisposed players to injury. PMID- 8711550 TI - South African geneticists' attitudes to the present Abortion and Sterilisation Act of 1975. AB - STUDY OBJECTIVE: To ascertain the attitudes of clinical geneticists and genetic counsellors practising in South Africa to the current Abortion and Sterilisation Act of 1975 (the Act). DESIGN: Postal questionnaire. MAIN RESULTS: Ninety-two per cent of the questionnaires were returned, and the responses were comparable to those of the South African Society of Obstetricians and Gynaecologists in 1990 and the Society of Psychiatrists of South Africa in 1992. No respondent felt that the Act should be more restrictive, and only 4 (17.4%) felt that it was acceptable in its present form. Section 3(1)(c) of the Act, which relates to termination of pregnancy (TOP) on genetic grounds, was acceptable in its present form to 13 respondents (56.5%), but 16 (69.7%) considered that TOP on genetic grounds should not be curtailed in future legislation. CONCLUSION: The geneticists' attitudes to the present Act concurred with those of the obstetricians and psychiatrists previously documented, and confirmed the need for review of the Act. All three specialist groups appeared to support an increase in the indications for legal TOP rather than the introduction of TOP on request up to a specified post-conceptual age. With regard to Section 3(1)(c), the geneticists' responses indicated an acceptance of the limitations of the present Act, coupled with concern about the implications of future changes to this section of the Act. PMID- 8711551 TI - The potential for preventing the delivery and perinatal mortality of low-birth weight babies in a black urban population. AB - OBJECTIVE: To determine the potential for preventing the delivery and perinatal mortality of low-birth-weight (LBW) babies in a black urban population. DESIGN: Cross-sectional descriptive study. SETTING: All women delivering babies weighing less than 2,500 g at Kalafong Hospital in a 6-month period (December 1991-May 1992). MAIN OUTCOME MEASURES: The primary obstetric reason for delivery; whether the labour was of spontaneous onset or iatrogenic; whether labour was theoretically preventable using currently accepted practice; the number of patients in whom suppression of delivery was attempted in the theoretically preventable group; and the perinatal mortality rate of that group. RESULTS: There were 124 perinatal deaths (22.5%) in the 550 LBW babies delivered from 465 singleton pregnancies, 42 twin pregnancies and 1 triplet pregnancy. The primary obstetric reasons for delivery were spontaneous preterm labour (28%), hypertensive diseases (19%), premature rupture of membranes (18%), spontaneous labour in light-for-gestational-age babies (16%), unexplained intra-uterine deaths (8%), antepartum haemorrhage (8%) and other causes (3%). A medical decision to terminate the pregnancy before labour was made in 177 (34.8%) cases, the major reason being hypertensive diseases (84 mothers; 47.5%). In the remaining 331 mothers with spontaneous onset of labour, labour was theoretically preventable in 63 (19%) and prevention was only attempted in 12 (2.4% of the total mothers). The major reason for not attempting to suppress labour in the others was that the patients arrived too late at the hospital for intervention to take place. CONCLUSION: Hospital staff can do little to prevent the delivery of LBW babies in a black urban population. PMID- 8711552 TI - Is there a place for radionuclide bone scintigraphy in the management of radiograph-negative scaphoid trauma? AB - OBJECTIVE: To evaluate the role of radionuclide bone scanning in patients with suspected scaphoid trauma, particularly in those with negative radiographs. DESIGN: Prospective. Radionuclide scans and carpal bone radiography were performed on all participants in the early post-injury period. SETTING: Cape Town tertiary centre trauma unit. PARTICIPANTS: Fifty patients who presented with clinical features suggestive of scaphoid trauma. MAIN OUTCOME MEASURE: Definitive radiographic diagnosis of fracture or persistent clinical features of scaphoid trauma. RESULTS: All patients who had fractures demonstrated on standard radiography either at the initial visit (13 patients) or at 2 weeks (8 patients) had positive scintiscans (sensitivity 100%). Four of 6 patients who had a positive scan but negative first and second radiographs had persistent tenderness on clinical examination which required extended immobilisation in a plaster cast. The overall positive predictive value of scintigraphy was 93%. All patients with a negative scan were clinically and radiologically negative at 2 weeks (negative predictive value 100%). Evidence of multifocal injury was present in 12 scans, but only 1 radiograph. Thirty-one patients (62%) were scanned within 48 hours of injury. CONCLUSION: Bone scintigraphy can be used in radiograph-negative scaphoid area injury to exclude the need for further follow-up reliably, but those with positive scans still require clinical examination and radiography at 2 weeks. PMID- 8711554 TI - The necessity for T-cryptantigen activation screening in babies with necrotising enterocolitis. AB - OBJECTIVE: To determine the prevalence of T-cryptantigen activation (TCA) and its predictive value for severity of necrotising enterocolitis (NEC) in babies. STUDY DESIGN: Prospective descriptive. STUDY POPULATION: Thirty-four babies with NEC were prospectively screened for TCA at Tygerberg Hospital over a 6-month period. TCA screening was done by testing for red blood cell agglutination by the common peanut lectin, Arachis hypogea. Once TCA was confirmed, only washed red cells were administered to the babies and plasma-containing blood products were avoided. NEC was divided into suspected NEC (stage 1), classic NEC (stage 2) and fulminant NEC (stage 3). MAIN OUTCOME MEASURES: Prevalence of TCA in babies with various stages of NEC; the association between TCA and bowel necrosis, need for surgery and mortality. RESULTS: TCA was positive in 8 (24%) of the babies in this study. Six babies (18%) had stage 1 NEC, 10 (29%) had stage 2 NEC and 18 (53%) had fulminating or stage 3 NEC. All 18 babies with stage 3 NEC required surgery and TCA was present in 8 (47%) of them. Twelve babies (35%) died, 3 with TCA and 9 with no TCA. Babies with TCA had portal venous gas on abdominal radiographs (P = 0.037) and stage 3 NEC (P = 0.003) more often than babies with no TCA. CONCLUSIONS: A strong association was noted between TCA and the fulminant form of NEC with bowel necrosis. TCA is a baby with NEC should alert the surgeon to the possibility of severe disease and the need to avoid plasma-containing blood products. Blood banks are urged to introduce routine screening for TCA in all babies with NEC. PMID- 8711553 TI - Intravenous immunoglobulin prophylaxis in neonates on artificial ventilation. AB - The efficacy of the prophylactic use of intravenous immunoglobulin (Ig) was evaluated in a double-blind placebo-controlled trial of 21 pairs of ventilated neonates weighing more than 1,500 g. Each infant received 0.4 g/kg/day of intravenous Ig or a similar volume of placebo daily for 5 days. Criteria used to assess the efficacy of intravenous Ig were the number of infections, the duration of ventilation therapy and time to clinical recovery. There were no significant differences in the treated and placebo groups with regard to the frequency of positive blood cultures (28.6% and 14.3%), endotracheal cultures (57.1% and 66.7%) and abnormal white cell counts (52.4% and 57.1%). On entry to the study there was no significant difference in IgG levels between the treated (974.5 mg/dl; SD 575.3) and placebo groups (818 mg/dl; SD 516.9). However, on day 6 the treated group had a mean level of 1,400.3 mg/dl (SD 426.7) versus 710.9 mg/dl (SD 377.4) in the placebo group (P < 0.05). Clinical improvement occurred within 3 days in both groups. Ventilatory support was required for 11.8 days (SD 8.3) in the treated and 11.8 days (SD 7.3) in the placebo group. Both groups required 3-4 antibiotic treatments over a period of 14-15 days. Two patients died in the treated and 4 in the placebo group, with 1 infant in each group developing bronchopulmonary dysplasia. The patients who recovered did so within 14 days. Analyses of subgroups of patients with different diagnoses revealed no differences except a trend suggesting fewer infections in term babies treated with intravenous Ig. The organisms cultured in the intravenous Ig groups were Pseudomonas, Klebsiella, Escherichia coli and Staphylococcus and in the placebo group Pseudomonas, Klebsiella and Enterobacter. The above has shown that, except for a trend in the older neonates, intravenous Ig is not of prophylactic benefit in ventilated neonates. Newer adjuncts in immunotherapy such as hyperimmune gammaglobulin or monoclonal antibodies may prove of greater value in the treatment of neonatal sepsis. PMID- 8711556 TI - A massive outbreak of food poisoning--a reminder of the importance of proper toxic waste control. AB - Because of rapid urbanisation in South Africa, scavenging from waste disposal sites by poor communities poses an increasing health risk. Reject cough lozenges, some of which contained larger amounts of dextromethorphan than usual, were illegally removed from a disposal site and, after resale by informal traders, caused moderately severe symptoms of toxicity in 171/540 (24%) primary school pupils. Although dextromethorphan was implicated as a cause, contributing effects of other toxins could not be excluded. Bacteriological cultures and a pesticide screen were negative. Had more toxic substances been involved, the consequences would have been disastrous. This incident supports calls for an integrated national waste management policy and waste control act to govern the management and control of waste from generation to disposal. Such a policy is necessary to prevent potentially serious incidents in the future. PMID- 8711555 TI - A phase II trial of fludarabine in patients with previously treated chronic lymphocytic leukaemia. AB - OBJECTIVES: To evaluate fludarabine in patients with chronic lymphocytic leukaemia (CLL) not responding to standard treatment. DESIGN SETTING: Fludarabine was administered for 5 consecutive days and repeated 4-weekly. SUBJECTS: Seventeen patients at a single institution were treated. OUTCOME MEASURES: Objective remission was seen in 11 patients. The median survival time was 356 days. RESULTS AND CONCLUSIONS: Fludarabine is an effective treatment for patients with advanced CLL. PMID- 8711557 TI - Tuberculosis of the cervical spine. AB - Tuberculosis of the cervical spine is rare, comprising 3-5% of cases of tuberculosis of the spine. Eight patients with tuberculosis of the cervical spine seen during 1989-1992 were reviewed. They all presented with neck pain. The 4 children presented with a kyphotic deformity. In all the children the disease was extensive, with a large prevertebral abscess formation, while in the adults it was localised to one or two motion segments. Cord compression was present in 4 of the 8 patients. All the patients were treated with antituberculosis drugs and 6 underwent surgery. There was full neurological recovery in all patients. The kyphosis was improved though not fully corrected. There was a problem in stabilisation of severe involvement of the body and dens of C2. Surgery seems to play a major role in the treatment of tuberculosis of the cervical spine. PMID- 8711558 TI - The humanitarian and scientific interests of Thomas Hodgkin (1798-1866), discoverer of a disease. AB - Thomas Hodgkin is generally famous for the discovery of a lymphoma in 1837, but not for his remarkable relationship as physician and friend with the philanthropist, Sir Moses Montefiore (1784-1885), and their six shared journeys to Europe, the Near East and North Africa to alleviate the plight of religious and ethnic minorities. Also less well known are Hodgkin's humanitarian activities, inspired by his being a Quaker: his assistance of freed slaves, and his involvement in the movement for the abolition of capital punishment and in mental hospital and prison reform. His wide range of scientific interests included medical education, geography, ethnology and social anthropology. He is buried in Jaffa, Israel, where he died of dysentery while on a trip with Montefiore. PMID- 8711559 TI - Occupational lung disease in ex-mineworkers--sound a further alarm! PMID- 8711560 TI - Tuberculosis Control Programme Practical Guidelines, 1996--treatment regimens. PMID- 8711561 TI - Tuberculosis control in South Africa--time for a new paradigm? PMID- 8711562 TI - Tuberculosis control in South Africa--time for a new paradigm? PMID- 8711563 TI - Cannabis--deceptive weed or decriminalised drug? PMID- 8711564 TI - Treatment of Acanthamoeba keratitis. PMID- 8711565 TI - Informed consent--a survey of doctors' practices in South Africa. PMID- 8711566 TI - [Revision of undergraduate curriculum]. PMID- 8711567 TI - Reasons for failure to administer antenatal corticosteroids in preterm labour. PMID- 8711568 TI - Multiple cigarette burn wounds in a chronic paranoid schizophrenic. PMID- 8711570 TI - The 64th congress of the European Atherosclerosis Society. Utrecht, The Netherlands, 10-13 June 1995. PMID- 8711569 TI - Health policy machinations: Manto Tshabalala. PMID- 8711571 TI - Myocardial hypertrophy--Part II. PMID- 8711572 TI - Dofetilide--medical and media information. PMID- 8711573 TI - Pravastatin improves outcome for heart transplant recipients. PMID- 8711574 TI - Progression of renal diseases--angiotensin-converting enzyme inhibitors versus calcium antagonists. PMID- 8711575 TI - Calcium channel blockers and associated risk of myocardial infarction. PMID- 8711576 TI - Long-acting calcium channel blockers not subject of FDA panel recommendation. PMID- 8711577 TI - Bisoprolol launched in South Africa. PMID- 8711578 TI - Infective endocarditis--what are the issues? PMID- 8711579 TI - Toronto stentless aortic valve replacement in elderly patients. AB - From July 1992 to October 1994, we inserted new Toronto SPV stentless aortic heterografts (SJM Med. Inc., St Paul, Minneapolis, USA) in 40 of a series of 50 consecutive patients older than 70 years. The mean age was 75.7 years (range 70 - 86 years). All, except 4 patients, were pre-operatively in NYHA functional class III or higher. The aortic clamp time was significantly higher in the stentless groups (75 v. 53 minutes, P < 0.001). The average 25.5 mm size of the implanted valves stands in stark contract to the low body surface area (1.69 m2) of this patient groups. The surgeon's (in)experience was the major reason for the drawbacks (5/50) associated with a stentless procedures. The follow-up period ranged from 2 to 27 months and was complete in 100% of cases. We encountered 1 hospital death and no late deaths (97.5% actuarial survival). The mean NYHA class at follow-up was 1.5, and without exception patients were in class I or II. We noted one transient ischaemic attack immediately postoperatively and another later incident in a patient with a previous severe vascular history. With a low intensity anticoagulation regiment for the first 3 months, there were two incidents of haemorrhaging necessitating premature anticoagulation withdrawal. Echocardiographic transthoracic valvular gradients compared favourably with the reported gradients of other biological valves, especially the smaller ones the significantly better haemodynamics were noted in most cases 6 months after implantation. Comparison of data with stented valves implanted during the same period indicates that the average size of the stentless valves was significantly higher (22.3 v. 25.5 mm, P < 0.001) in an equivalent population. PMID- 8711580 TI - Atypical clinical presentation of bileaflet cardiac prosthetic valve dysfunction. Preliminary report. AB - Single-leaflet disc immobilisation in 3 patients with St Jude Medical prostheses and 1 patient with a Carbo Medics valve prosthesis resulted in mild mitral regurgitation without acute pulmonary oedema. Three patients presented with slowly worsening pulmonary congestion while 1 patient had neurological symptoms. Clinicians need to be aware of the possibility of single-leaflet failure in a bileaflet cardiac valve prosthesis, because haemodynamic deterioration occurs insidiously in these patients, resulting in atypical symptoms and signs. PMID- 8711581 TI - Arrhythmias induced by exercise in athletes and others. AB - Athletes are subject to the same arrhythmias as the general population, but the frequency and significance of the arrhythmias may be different. Cardiovascular conditioning slows the heart rate and may make athletes more vulnerable to neurocardiogenic syncope and atrial fibrillation. Tachyarrhythmias may be precipitated by vigorous exercise and more severe rate-related symptoms may result because of the high sympathetic drive during sports activities. For those with pre-existing cardiovascular abnormalities, athletic activity may be beneficial in some cases, but dangerous and even life-threatening in others. A review of the subject and recommendations based on our personal experience and a recent consensus conference are provided. PMID- 8711582 TI - Comparison of the effects of propofol and halothane on acute myocardial ischaemia and myocardial reperfusion injury. AB - The effects of two blood concentrations of propofol on acute severe regional myocardial ischaemia (15-minute occlusion of the LAD coronary artery) and 2-hour reperfusion were studied in 10 pigs. Results were contrasted with the effect of 0.8% halothane (N = 5). Results indicate that during acute ischaemia, regional myocardial function of the segment supplied by the LAD artery was not protected against the effects of ischaemia in the propofol or halothane groups. During reperfusion, animals subjected to halothane demonstrated significantly fewer reperfusion arrhythmias and segmental function was less stunned than in either propofol group. There was no difference in heart performance between the propofol groups. Results indicate that the proposed anti-oxidant effect of propofol is not sufficient to protect the animal heart from reperfusion injury. PMID- 8711583 TI - The haemodynamic correlations of the cardiopulmonary flow index, a relatively non invasive technique for diagnosis of myocardial pump insufficiency. AB - The cardiopulmonary flow index (CPFI) is the ratio of cardiopulmonary blood volume to stroke volume. OBJECTIVE. To compare the CPFI derived from a first-pass radiocardiogram and electrocardiogram with the usual haemodynamic measurements obtained by bedside catheterisation. DESIGN. Sixty-one patients in a coronary care unit who had flow-directed pulmonary artery catheterisation for clinical indications had a CPFI recorded together with the usual haemodynamic measurements and calculations. RESULTS. In patients with myocardial pump failure without significant pulmonary disease, linear regression analysis indicated that the best fit for the CPFI could be obtained with a model that used the inverse of stroke index and pulmonary capillary wedge pressure as independent variables. Ninety-six per cent of the variation in CPFI can be explained by variations in pulmonary capillary wedge pressure and stroke index. The CPFI correlated less well with these variables in patients with concomitant lung disease. CONCLUSION. The CPFI is a potentially useful and relatively non-invasive index of haemodynamic function in acutely ill patients without significant pulmonary disease. PMID- 8711584 TI - The switch to balloon atrial septostomy with echocardiographic guidance--a severe complication. AB - To warn about a severe complication of balloon atrial septostomy (BAS) guided by echocardiography, a retrospective review was undertaken of all BASs performed at Red Cross War Memorial Children's Hospital in Cape Town from 1984 to the end of 1992. BAS with fluoroscopy was carried out in 41 infants (group I). Since April 1991, 17 infants had BAS with two-dimensional echocardiography guidance in the intensive care unit (group II). The majority of patients (72%) had simple transposition of the great arteries. Only 3 patients in group I were catheterised via the umbilical vein (the remaining 38 via the femoral vein). In group II, 12 patients (71%) were catheterised via the umbilicus. The only fatality as a result of the procedure occurred in group II. This was the result of rupture of the splenic vein. The patient was catheterised via the umbilical vein, and echocardiography was not adequately used to guide the catheter below the diaphragm. There were no other severe complications in either group. Although echocardiographically guided BAS is easier, quicker, potentially safer and keeps the baby in a better homeostatic environment, we describe a severe complication of this procedure in the learning phase that has not been noted previously. PMID- 8711585 TI - [PSA and prostatic adenocarcinoma. Experience with 805 patients]. AB - Over the last decade PSA has been widely acknowledged to be a very sensitive and specific marker of prostatic tumour pathologies. This paper reports the authors' experience relating to the use of PSA in the early diagnosis of prostatic heteroplasia on the basis of results obtained in a study performed in 805 dysuric patients who underwent rectal exploration and CAT. Echo-guided prostate biopsy according to Hodge was performed in 212 (26.4%) out of 805 patients and adenocarcinoma was found in 55 cases (25.9%). PSA ranged between 10-198 ng/ml in 42 out of 212 patients, between 4-10 ng/ml in 60, and was below 4 ng/ml in 110. In addition, PSAd was assayed in all patients with PSA < 10 mg/ml. Having established the cutoff of PSA at 10 ng/ml, it was found that some heteroplasia, above all at the initial sage, presented normal PSA blood values. In fact, in 8 cases, equivalent to 14.5% of the neoplasias diagnosed, values were under 4 ng/ml, and in 5 cases, equivalent to 9%, they ranged between 4-10 ng/ml. This finding showed that the threshold value of 10 ng/ml gives PSA a high specificity, but a lower sensitivity in the early diagnosis of prostatic adenocarcinoma. Moreover, the analysis of PSA-density does not significantly allow the diagnosis of a larger number of heteroplasias: in fact, out of 13 cases of adenocarcinoma with PSA < 10 ng/ml, PSAd was only > 0.15 in 2 patients. Therefore, in line with the data reported in the literature, the authors consider that PSA assay represents a sensitive and specific screening method for prostatic tumour pathologies in symptomatic patients, but in order to obtain an early diagnosis, and especially in cases with serum values between 4-10 ng/ml, it must be combined with both rectal exploration and, above all, transrectal scan. PMID- 8711586 TI - [The role of TPS in the diagnosis of prostatic carcinoma]. AB - From June 1993 to July 1994 we treated 136 male patients affected by dysuria with hematic dosage of TPS and PSA, with DRE, transrectal echography and prostatic biopsy using an echograph. The patients treated were divided into two groups: A and B. Group A (TPS cut-off of 75 U/l) was made up of 91 patients, ages ranging from 41 to 85 years (average 72.5 years). Group B (TPS cut-off > 75 U/l) was made up of 45 patients, ages ranging from 39 to 88 years (average 75.1 years). In group A, 22 patients (24.2%) resulted as suffering from prostatic carcinoma, 4 (4.3%) resulted as suffering from prostatic phlogosis, and 65 (71.4%) resulted as suffering from BPH. In group B, 6 (13.3%) were affected by prostatic heteroplasia, 2 (4.4%) were affected by prostatic phlogosis, and 37 (82.2%) were affected by BPH. The TPS had a sensibility of 21.4% and a specificity of 66.3%. A comparison of the data obtained in the two groups did not reveal any significant differences. In this study the TPS showed a poor diagnostic reliability as a biologic marker of prostatic carcinoma. PMID- 8711587 TI - [Natural history of prostatic adenocarcinoma. Recent advances in prognostic parameters and biological behavior]. AB - For many years diagnosis and management of prostatic adenocarcinoma were straightforward. However, with the advent of medical and less invasive therapies for the treatment of prostatic cancer, it has become necessary to know more about the etiology and pathophysiology of the disease. For example, hormonal factors, growth factors, stromal-epithelial interactions and aging have all been implicated in the etiology of the disease. The pathology of the disease demonstrates heterogeneity in the ratio of the stroma to the epithelium in any given patient, and the pathophysiology varies. Recognizing this heterogeneity in the disease, it is clear that no one form of medical or interventional therapy should be expected to result in a complete response. PMID- 8711588 TI - Is medical therapy an expensive way for delaying surgery in BPH patients? AB - Nowadays, no medical therapy can be considered as a real and definitive alternative to surgery in the management of BPH patients. We considered pharmacologic approach as a treatment that may delay the need for surgery for BPH. In some cases a delayed therapy may continue for all patient life, excluding the need for TURP. The questions that we propose in the present review are: Is there always a role for a delayed medical therapy in the treatment of BPH patients? In which BPH patients a delayed medical therapy and in which instead an immediate surgery may be chosen? Which factors may influence this decision? A delayed medical therapy cannot be chosen in all BPH cases. Two factors can influence the evolution of the disease and the decision of the therapy: the first, natural history of BPH is related to BPH progression, and the second to patient characteristics. The role of growth factors in the natural history of BPH is investigated. Age of patient, his health condition and the presence of concomitant diseases are characteristics that may influence the therapeutic choice. In a young patient with good health condition and no concomitant diseases, the specific clinical phase of BPH is crucial to determinate the need for surgery of for medical delayed therapy. If there is a worsening health status or concomitant diseases as diabetes and hypertension that can increase the risk related to surgery in the future or can determine a more rapid evolution of BPH, TURP may be immediately recommended in all clinical phases of prostatic hyperplasia. The role of age in this therapeutic decision must be carefully examined. PMID- 8711589 TI - [A case of adrenal neoplasm]. AB - Pheocromocytoma is a rare adrenal tumor. Generally its size is reduced and it is one of the causes of arterial hypertension. The authors present a case of giant pheocromocytoma, without arterial hypertension, and particularly they write about the used diagnostic and therapeutic management. PMID- 8711590 TI - [Large inguino-scrotal bladder herniation]. AB - The authors describe a case of voluminous inguinoscrotal bladder erniation. They write about the subjet with the help of the literature and above all they show the possible dangers of a vesical underestimated injury. PMID- 8711591 TI - [Rupture of the upper urinary tract during renal colic. Two cases]. AB - The rupture of the upper urinary tract is an uncommon event. The authors describe 2 cases during renal colic and they write, particularly, about the diagnostic and therapeutic aspects of this pathology. PMID- 8711592 TI - [Carcinoma of the prostate, stage C]. PMID- 8711593 TI - [Medical therapy of locally advanced prostatic neoplasms]. PMID- 8711594 TI - [What is the role of neoadjuvant therapy in prostate cancer?]. PMID- 8711595 TI - [Comments on radical prostatectomy three month after hormone therapy]. PMID- 8711596 TI - Colectomy. The evolving role of laparoscopy. PMID- 8711597 TI - Laparoscopic colectomy vs traditional colectomy for diverticulitis. Outcome and costs. AB - BACKGROUND: The aim of this study was to evaluate the outcome of patients undergoing laparoscopic colectomy for diverticulitis. METHODS: Fourteen consecutive patients undergoing laparoscopic sigmoid colectomy (LSC) for diverticulitis were evaluated. Medical records from a control group of 14 matched patients undergoing traditional open sigmoid colectomy (OSC) for diverticulitis were reviewed for comparison. RESULTS: Mean age, operative time, morbidity, and mortality of the LSC and OSC groups were not significantly different. However, the mean estimated blood loss (171cc vs 321cc), days to p.o. liquids (2.9 vs 6.1), and postoperative stay (6.3 vs 9.2 days) were all significantly less in the LSC patients. Although the mean operating room charges were greater in the LSC patients ($10,589 vs $8,207) the mean total hospital charges ($29,981 vs $36,745) and costs ($11,528 vs $13,426) were markedly less. CONCLUSIONS: Compared with OSC for diverticulitis, LSC results in a more rapid return of bowel function and shortened hospital stay. Despite the greater operating room charges of LSC, the total hospital charges and costs are lessened. PMID- 8711598 TI - Laparoscopic evaluation of penetrating thoracoabdominal traumatic injuries. AB - BACKGROUND: General surgeons' recent familiarity with advanced laparoscopic techniques have rendered laparoscopy feasible safely in the trauma setting. Traditionally high rates of nontherapeutic laparotomies also contribute to this increased interest. This study was undertaken to determine the predictive value and accuracy of diagnostic laparoscopy (DL) in evaluation of penetrating thoracoabdominal trauma. METHODS: Entry criteria included thoracoabdominal gunshot (GSW) or stab wounds (SW) in otherwise hemodynamically stable patients. A high index of suspicion for either hemoperitoneum, peritonitis, or diaphragmatic injury was required for inclusion. All patients underwent DL in the operating room followed by standard laparotomy. The findings of the two evaluations were compared. RESULTS: Twenty-four patients were included in the study. Twenty males and 4 females with an average age of 34 years made up the group. Violation of the peritoneal cavity was present in 21 cases and absent in 3. No intraabdominal injuries were found during laparotomy in the latter three cases without peritoneal violation. The specificity and positive predictive value were 100% for lesions of the diaphragm, liver, spleen, pancreas, kidney, and hollow viscus. The sensitivity was highest for liver and spleen injuries (88%), followed by diaphragmatic injuries (83%), pancreas and kidney injuries (50%), and lowest for injuries of hollow viscus (25%). The negative predictive value was 95, 99, 91, and 57%, respectively, for these organs. CONCLUSIONS: DL could have avoided unnecessary laparotomy in 38% of cases in this study. There were no complications related to laparoscopy. The greatest value of DL in penetrating thoracoabdominal injuries is in the evaluation of peritoneal violation, diaphragmatic, and upper abdominal solid-organ injuries. It is not ideal for predicting hollow viscus injuries. PMID- 8711599 TI - Teaching basic video skills as an aid in laparoscopic suturing. AB - BACKGROUND: There is a perception among surgeons that performing laparoscopic suturing is unduly difficult. The purpose of this study is to document a program which aides in learning laparoscopic suturing. METHODS: Fourteen volunteer medical students without prior experience were taught laparoscopic suturing. Videoscopic pelvitrainers were utilized for a 2-h training session. Extracorporeal and intracorporeal knot tying was demonstrated utilizing a three throw square knot. After a 2-h practice session each student's time to complete an extracorporeal and intracorporeal suture was recorded. RESULTS: The average times required for completion were: extracorporeal suture and knot 1 min 54 s; intracorporeal suture and knot 3 min 12 s. CONCLUSIONS: Novice students were able to perform at extra and intracorporeal suturing with 2 h of practice, utilizing a systematic program of teaching basic video skills. PMID- 8711600 TI - Resident education in laparoscopic cholecystectomy. AB - BACKGROUND: Resident education in laparoscopic cholecystectomy (LC) was studied in a retrospective analysis of consecutive cases performed at two academic institutions with different educational approaches. METHODS: Each procedure was performed by a resident as operating surgeon under the direct guidance of one of a small, constant group of LC-certified attendings acting as first assistant. In group I (n = 48), residents acquired LC skills by graded exposure and surgical responsibility similar to their training in other general surgical procedures. In group II (n = 48) residents were additionally certified via an intensive course (including didactic and animal model experience) prior to assuming responsibility as surgeon. RESULTS: Results were similar in each group. No technical errors were identified. Blood transfusion was not required related to surgery. Conversion to an open procedure occurred in 10% and 8% in groups I and II, respectively. The rate of complications was 4% for group I and 8% for group II. A longer operating time was noted in group I and may be attributed to nonoperative reasons. CONCLUSIONS: Education in LC via graded experience throughout residency achieves results similar to that found with the addition of an intensive course. This additional training may not be necessary for surgical residents. PMID- 8711601 TI - A comparison of the approaches to laparoscopic herniorrhaphy. AB - BACKGROUND: There are a variety of accepted techniques for herniorrhaphy. With the advent of laparoscopic general surgery, new endoscopic techniques using the transabdominal and total extraperitoneal approaches have been added to the many options for the repair of inguinal hernia. The purpose of this study was to compare the early results of these approaches at a single institution. METHODS: Between May 1991 and August 1994, 600 laparoscopic herniorrhaphies were performed on 493 patients. Three hundred hernias were repaired using the transabdominal preperitoneal approach and 300 were repaired using the total extraperitoneal approach. A retrospective review was performed with emphasis on the comparison of recurrence rates and complication rates between these two approaches to laparoscopic herniorrhaphy. RESULTS: The recurrence rates were 2.0% (6/300) for the transabdominal approach and 0.3% (1/300) for the total extraperitoneal approach. The complication rate for the transabdominal approach was 10.7% and included thigh paresthesias (6), inferior epigastric artery injuries (4), enterotomy (1), bowel obstruction (1), bladder injury (1), and urinary retention (14). The complication rate for the total extraperitoneal approach was 3.7% and included enterotomies (2), bladder injury (1), paresthesia (1), and urinary retention (6). The recurrence, the enterotomies, and the bladder injury in the total extraperitoneal group were all in patients who had previous lower abdominal operations. CONCLUSIONS: Although both the transabdominal preperitoneal and total extraperitoneal approaches to laparoscopic herniorrhaphy have acceptable recurrence and complication rates, there were significant advantages to the total extraperitoneal approach in our institution. Previous lower abdominal surgery may be a relative contraindication to the total extraperitoneal approach. PMID- 8711602 TI - Laparoscopic cholecystectomy in the elderly. AB - BACKGROUND: Few studies have examined the results of laparoscopic cholecystectomy (LC) in the elderly. We reviewed our experience with the procedure in 194 patients age 65 and older. METHODS: A chart review was performed on patients who underwent attempted LC over a 4-year period. Age, conversion rate to open cholecystectomy (OC), length of stay, and morbidity and mortality rates were compared between elective and inpatients as well as between patients age 65-75 and patients over age 75. RESULTS: Conversion rate to OC was 10.6%. Mean length of hospital stay was 2.7 days. Morbidity and mortality rates were 18% and 1%. Elective patients experienced significantly fewer medical complications. There were no differences in complication rates between patients age 65-75 and patients over 75 years, but younger patients had a significantly shorter mean length of hospitalization. CONCLUSIONS: Elderly patients experience more complications and longer lengths of stay than the general population. However, our results compare favorably with OC series in elderly patients. PMID- 8711603 TI - ERCP in association with laparoscopic cholecystectomy. A strategy to minimize the number of unnecessary ERCPs. AB - BACKGROUND: With the evolution of laparoscopic cholecystectomy (LC) as the standard operation for benign gallbladder disease, the role of endoscopic retrograde cholangiopancreatography (ERCP) in the management of common bile duct (CBD) stones has to be defined. METHODS: From November 1990 to April 1994 we attempted LC in 1,788 patients. Eighty-nine patients underwent ERCP preoperatively under the following indications: jaundice or a history of jaundice, cholangitis, gallstone pancreatitis, abnormal liver function tests, and a sonogram showing either CBD stones or a dilated CBD. With intent to minimize the number of unnecessary ERCPs only patients with jaundice, cholangitis, and high abnormalities on the liver function tests (LFTs) were directly referred for ERCP. All other patients with suspected choledocholithiasis were initially investigated with intravenous cholangiography (IVC) and tomography; only patients with positive findings on IVC subsequently underwent ERCP. Eighteen patients underwent ERCP postoperatively and the indications included jaundice, bile leak, and abnormal intraoperative cholangiogram. RESULTS: Of the 89 patients having ERCP preoperatively 54 patients (60.7%) were found to have CBD stones which were removed endoscopically in all cases except in one patient where a large CBD stone was removed during laparoscopic exploration of the CBD. Eight patients of the 18 patients having ERCP postoperatively were found to have CBD stones and all of them had their CBD cleared endoscopically. There were no mortalities, while four patients developed a mild pancreatitis. CONCLUSIONS: Although there is an increasing tendency to clear the bile duct with a laparoscopic approach, ERCP and sphincterotomy has a certain role in conjunction with LC in the management of patients with a high suspicion of CBD stones, particularly in institutions where there is easy access to expert interventional endoscopic techniques. PMID- 8711604 TI - Exposure for laparoscopic cholecystectomy dissection adversely alters biliary ductal anatomy. AB - BACKGROUND: Exposure for open cholecystectomy entails lateral, caudal traction on the gallbladder infundibulum, which results in opening the angle between the cystic and hepatic ducts. Laparoscopic cholecystectomy (LC), as initially described, is done with cephalad traction on the gallbladder. We hypothesized LC exposure technique narrows the angle between the cystic and hepatic ducts, placing them at increased risk of injury. METHODS: Twenty-three patients had routine LC. Cystic duct cholangiography (IOC) was done with a flexible 5-Fr catheter via a percutaneous introducer placed anterior to the gallbladder. Exposure of Calot's triangle was maintained with cephalad traction on the gallbladder fundus. IOC was repeated after allowing the organ to assume the anatomic position. The cholangiograms were inspected for significant differences, and the angle of the cystic to the hepatic duct (CDHD) was measured by a blinded radiologist. RESULTS: The mean angle of the cystic to hepatic duct was 30 degrees +/- 19 degrees in the IOCs taken with cephalad traction on the gallbladder fundus vs 59 degrees +/- 22 degrees, P < 0.001, in the cholangiograms taken without traction. A filling defect at the cystic-hepatic duct junction was present in 39% of IOC taken with traction vs none without traction. The intrahepatic ducts were seen in all films without traction, whereas the intrahepatic ducts were not visualized in 13% of IOCs taken with traction. CONCLUSIONS: From these data we conclude (1) extra-hepatic biliary ducts may be at increased risk of injury during LC because of the exposure technique and (2) imaging bile ducts in the anatomic position may convey misleading information about the relative location of important structures. Optimal exposure for dissection of Calot's triangle should utilize a second clamp on the infundibulum with lateral, caudal traction. PMID- 8711605 TI - A double-blinded evaluation of intraperitoneal bupivacaine vs saline for the reduction of postoperative pain and nausea after laparoscopic cholecystectomy. AB - BACKGROUND: Intraperitoneal local anesthesia has been reported to reduce postoperative pain after laparoscopy for gynecologic procedures that do not require a great deal of dissection or manipulation of viscera. This study was performed to determine the efficacy of intraperitoneal bupivacaine in laparoscopic cholecystectomy (LC). METHODS: Fifty-five patients were evaluable in this randomized, double-blind, placebo-controlled study. Twenty-six patients received bupivacaine (0.1%) and 29 patients received placebo (saline). Prior to any dissection of the gallbladder, the surgeon irrigated 100 ml of experimental solution under the right hemidiaphragm, over Glisson's capsule, over the gallbladder serosa, and into the subhepatic space. The operation was then performed as usual. Postoperatively, analgesic medication usage, nausea, vomiting, and pain scores were determined during hospitalization. A questionnaire was given to each patient upon discharge from the hospital in order to continue monitoring medications and pain for the first 48 h at home. RESULTS: Postoperative pain was reduced significantly (P < 0.05) in the patients who received bupivacaine, but the effect was modest and observable only during the first 6 h after surgery. Despite this difference, there was no significant reduction in the amount of analgesic medication used by the patients who received bupivacaine, nor was there any reduction in nausea, vomiting, or shoulder pain when queried specifically. CONCLUSIONS: Intraperitoneal bupivacaine offered a detectable, albeit subtle benefit to patients undergoing LC. However, the effect was transient and had little impact upon the patient's convalescence. PMID- 8711606 TI - Absorbable clips for cystic duct ligation in laparoscopic cholecystectomy. AB - BACKGROUND: The efficacy and applicability of an absorbable polydioxanone (PDS) clip for cystic duct ligation were evaluated in 297 patients undergoing laparoscopic cholecystectomy. METHODS: The indications for cholecystectomy were symptomatic gallstones (179 patients), acute cholecystitis (67), biliary pancreatitis (23), acute cholangitis (24), and gallbladder polyp (4). RESULTS: Twenty-five patients required conversion to open surgery (8.4%). The conversion rate was 2.7% for uncomplicated and 17.5% for complicated gallbladder diseases. Of the 272 patients with laparoscopic cholecystectomy, the cystic ducts were successfully ligated with PDS clips in 227 patients (83.5%). The success rate was higher in uncomplicated (163/178) than in complicated (64/94) gallbladder diseases (chi square = 24.6, P < 0.001). There was no clip-related complication on follow-up (range 0.4-39.2, median 17.5 months). In 45 patients, PDS clip failed. They were treated with endoloop (14 patients), Roeder slip knot (13), metallic clips and endoloop (8), metallic clips alone (6), and intracorporeal tie (4). CONCLUSIONS: The PDS clip is effective and applicable to the majority of patients. It should be attempted first because of the ease of application. PMID- 8711607 TI - 149 ambulatory laparoscopic cholecystectomies. AB - BACKGROUND: The purpose of this study was to compare ambulatory laparoscopic cholecystectomy (LC) with overnight LC and to determine its safety and the factors which will predict its success. METHODS: Ambulatory LC was defined as LC followed by less than a 12-h stay in the ambulatory surgery unit; overnight LC was defined as LC followed by a hospital admission of less than 24 h. A retrospective chart review was completed. RESULTS: One hundred forty-nine ambulatory LC, were performed in an 18-month period. Ninety-one were successful (61%); 58 patients required overnight admission for pain management, control of nausea, or their reluctance to be discharged. There were five complications and no mortalities. The duration of the surgical procedure was the only significant objective factor we could find in predicting success of ambulatory LC. CONCLUSIONS: We conclude that ambulatory LC is safe and effective in treating patients requiring cholecystectomy. The duration of the procedure and the patient's own motivation are key factors in predicting success of early discharge. PMID- 8711608 TI - Results of laparoscopic treatment of hydatid cysts of the liver. AB - BACKGROUND: We have previously reported on the laparoscopic treatment of hydatid cysts of the liver. We now report the successful treatment of 18 cysts in 12 patients with a median follow-up of 12 months. METHODS: The standard treatment in the open technique was performed laparoscopically. The main concern was to prevent spillage of the hydatid fluid, for which the puncture and evacuation of the cyst is carried under scolicidal agents cover using 1% cetrimide, thus decreasing the chances of recurrence. RESULTS: In this group of patients we had one biliary leak that ceased spontaneously and one recurrence of hydatid cyst but not in the same lobe. CONCLUSIONS: We conclude that laparoscopic evacuation of hydatid cysts is a successful operation comparable to the open technique, with the added advantages of the laparoscopic approach. PMID- 8711609 TI - Peritoneal cystic mesothelioma treated with minimally invasive approach. AB - A case of peritoneal cystic mesothelioma in a young woman was treated with a minimally invasive approach. The technique and a review of the literature are presented. PMID- 8711610 TI - Gastric bezoar following penetrating abdominal injury. Diagnosis and endoscopic therapy. AB - We present a patient who developed a gastric bezoar following extensive penetrating abdominal trauma. Bezoar pathogenesis, diagnosis, and endoscopic therapy are highlighted. Additionally, alternative therapeutic modalities are explored with a review of the literature. Combination endoscopic lavage fragmentation/extraction presents a safe method of bezoar resolution. PMID- 8711611 TI - Thoracoscopic splanchnicectomy for the relief of intractable abdominal pain. AB - Video-thoracoscopic transthoracic splanchnicectomy has been applied to patients in the end stage of pancreas cancer who had intractable pain mediated through the splanchnic nerve in the left upper quadrant. The procedure is performed under general anesthesia in a right hemilateral position. Following the establishment of access to the thoracic cavity, the left splanchnic nerve is cut off at the level immediately above the aortic hiatus, through a small opening made in the pleura between the descending aorta and the vertebrae. All patients had immediate and complete relief of pain postoperatively. Only a transient drop in the mean arterial pressure was observed immediately after cutting off the nerve. No other detrimental effect of the procedure on the general condition was observed. No patients developed postoperative complications. The present method may, thus, be a treatment of choice directed toward the relief of intractable abdominal pain in selected patients with pancreatic cancer. PMID- 8711612 TI - Two techniques of creating periumbilical incisions during laparoscopic surgery. AB - Insertion of a periumbilical port is an integral part of laparoscopic surgery. It is often difficult to avoid bevelling of the skin when the semicircular periumbilical incision is made. We used two simple techniques for creating a uniform periumbilical incision. These methods were used in 128 consecutive laparoscopic cholecystectomies without complications. Achieving good cosmetic results is an important aspect of minimal access surgery. The techniques described have been successfully applied to laparoscopic surgery and easily taught to surgical trainees. PMID- 8711613 TI - New technology for the division of short gastric vessels during laparoscopic Nissen fundoplication. A prospective randomized trial. AB - Division of the short gastric vessels (SGV) is a standard component of laparoscopic Nissen fundoplications (LNF) at our institution. This study compares our original method of vessel control, multifire clip applier (MCA) and sharp division, to the Ultracision Harmonic Scalpel LCS (LCS). Twenty consecutive patients were evaluated in a randomized prospective fashion. Times for SGV division and estimated blood loss (EBL) were recorded. Cost data represent patient charges for use of either the MCA or LCS and the charge for operative time. Use of the LCS produced a significant reduction in the time required for SGV division and in the charges to the patient. [table: see text] We feel it is important to divide the SGV in all LNFs, and LCS use can result in significant savings of time and money. PMID- 8711614 TI - Three-dimensional ultrasound. AB - Laparoendoscopic surgery is fueled by technology, and advances in medical imaging may bring about further progress. The application of three-dimensional ultrasound to minimally invasive surgery represents technology "just over the horizon." This article describes the scientific basis of three-dimensional ultrasound and its ability "to see" anatomy not readily visualized laparoscopically. Three dimensional ultrasound may offer a more intuitive, accurate assessment of hidden anatomic structures in real time, using a safe, non-ionizing and cost-effective technology. PMID- 8711615 TI - "Dropped" surgical clips following laparoscopic cholecystectomy. PMID- 8711616 TI - A hypogastric approach to laparoscopic cholecystectomy. PMID- 8711617 TI - Hartmann's pouch, the Hartmann operation, the Hartmann procedure. An enigma of terminology. PMID- 8711618 TI - Inhibitors of glycolytic metabolism affect neurulation-staged mouse conceptuses in vitro. AB - In order to evaluate the apparent discordance between altered glucose metabolism and embryonic energy production, the effects of inhibitors of glucose utilization on morphological development and biochemical changes in mouse embryos in culture were evaluated. Day 9 ICR mouse conceptuses having 3-6 pairs of somites were prepared for culture as previously described. 2-Deoxyglucose (2DG) produced a concentration-dependent effect on development. A 25 microM 2DG concentration did not induce neural tube closure defects (NTDs) but 100 microM, 100% of embryos exhibited this defect. A 17% reduction in the rate of lactate production by the conceptus was produced by a 24-hr exposure period to 100 microM 2DG. Iodoacetate, which inhibits glyceraldehyde-3-phosphate dehydrogenase in adult tissues, produced high rates of NTDs at concentrations > or = 2.5 microM. Following a 24 hour exposure to iodoacetate, lactate production was inhibited at 10 and 25 microM. The effects of 2DG on embryonic ATP content were assessed to test the hypothesis that effects on glucose utilization would effect embryonic ATP content. Despite using 2DG concentrations that alter development and inhibit glycolysis, there were no effects on whole embryo or visceral yolk sac (VYS) ATP content. However, when the embryo was divided into regions, there was a specific reduction in ATP content in the head following a 24-hr exposure period. No effect of 2DG on head ATP content was produced after 12 hr of exposure. To determine if there were region specific differences in 2DG uptake and distribution that could account for the differential effects of 2DG on ATP content, 14C-2DG accumulation in different regions of the embryo and VYS was determined over the 24-hr culture period. The uptake of 2DG was dependent on the medium 2DG concentration and suggested a higher accumulation in regions with decreased ATP. However, when the uptake was monitored for a 1-hr period after a 24-hr exposure, there was no region specific differences in 2DG uptake. These studies further document the adverse developmental effects of inhibitors of glucose utilization during the early stage of neurulation. The biochemical mechanism for induction of these defects is unclear, but an effect on ATP content does not appear to be solely responsible for the dysmorphogenesis. PMID- 8711619 TI - Effect of treatment with phenobarbital and stiripentol on carbamazepine-induced teratogenicity and reactive metabolite formation. AB - A model using SWV mice was developed to investigate the mechanistic basis of carbamazepine (CBZ)-related fetotoxicity. Drug administration was initiated prior to conception and continued until day 18 of gestation. The incidence of malformation was 33% following CBZ exposure (1,500 mg/kg/day), compared with a 5% incidence in pair-fed control animals (P < 0.05). Coadministration of nonteratogenic doses of phenobarbital (PB; a cytochrome P-450 inducer) (45 mg/kg/day) and CBZ (1,000 mg/kg/day) increased the frequency of malformation from 10% to 26% (P < 0.05), compared with mice dosed with CBZ alone (1,000 mg/kg/day). Coadministration of stiripentol (STP; a cytochrome P-450 inhibitor) (300 mg/kg/day) decreased the incidence of malformations produced by CBZ (1,500 mg/kg/day) from 33% to 16.7% (P < 0.05). The effect of PB administration on the binding of 14C in maternal and fetal tissue was assessed in dams that received CBZ (1,000 mg/kg/day) with or without PB (45 mg/kg/day) or STP (300 mg/kg/day) chronically and a single i.p. dose of 14C-CBZ on day 12 of gestation. In all instances, binding was greatest in maternal liver, then in the placenta, fetal head and body, and maternal thigh muscle. In all tissues, PB caused a two-to threefold increase in binding, compared with binding in mice exposed to CBZ alone. STP administration decreased protein adduct formation only in maternal liver. The binding of 14C was also assessed in hepatic microsomes prepared from female mice exposed to CBZ and PB or STP as in the in vivo study of 14C binding. The extent of irreversible binding was 67% greater in microsomes prepared from mice pretreated with PB and CBZ than with CBZ alone, while STP resulted in only 21% inhibition of 14C adduct formation (P < 0.05). The results are consistent with the formation of a chemically reactive teratogenic metabolite of CBZ in mice by cytochrome(s) P-450. PMID- 8711620 TI - Functional involvement of Pax-1 in somite development: somite dysmorphogenesis in chick embryos treated with Pax-1 paired-box antisense oligodeoxynucleotide. AB - The metameric pattern of the vertebrate axial skeleton, defined by structures such as the vertebral bodies and ribs, is a result of segmentation events that occur during embryogenesis. The key event in axial segmentation is somite formation. This study examines the role of Pax-1, a member of the paired-box containing Pax gene family, in chick somite development. To investigate whether misexpression of Pax-1 during somite development is functionally related to abnormal axial patterning, antisense methodologies were used to perturb Pax-1 expression. An antisense, phosphorothioate-modified oligodeoxynucleotide (ODN) was designed based on the mouse Pax-1 paired-box sequence, and was either injected into or directly applied topically to early, somitic stage chick embryos. Abnormalities in somite structure and pattern were subsequently observed and scored, including loss of somites (18% of injected embryos, 35% of embryos treated by topical application), fusion of somites (25% of injected, 6% with topical application), and shortened body axis (0% of injected, 11% with topical application). Control embryos receiving sense ODN or non-sense ODN (a scrambled sequence with base composition identical to the antisense ODN) showed substantially fewer somite anomalies, indicating that the effects were sequence specific. These developmental abnormalities were analyzed using standard histological methods. Whole mount in situ hybridization was carried out to analyze the three-dimensional pattern of Pax-1 expression in whole embryos. In control, untreated embryos, the expression was localized to the entire epithelial somite, and as the somite matured, the expression was limited to its ventromedial region. With Pax-1 antisense ODN treatment, embryos with fused somites retained expression over the entire fused somite, and embryos that had complete loss of somites had greatly reduced expression of Pax-1 transcript. The results presented here provide strong evidence that Pax-1 is functionally important during somitogenesis and morphogenesis of the vertebral column. The spatial pattern of gene expression appears to delineate different populations of cells in the developing embryo (i.e., somite from somite, sclerotome from dermomyotome), and is consistent with the hypothesis that Pax-1 is involved in forming or maintaining boundaries at specific times and locations during development. PMID- 8711621 TI - Rachipagus conjoined twins: they really do occur! AB - The myth of rachipagus twins is hereby exploded-the myth, not the rachipagus! In a review of over 1,200 cases of conjoined twins, one classic example of rachipagus was found-two complete infants with dorsal union of the entire head and trunk, with roentgenograms showing clearly visible bony union of the vertebral arches from T6 to L3. After the typical case was discovered, 20 dorsal parasitic twins were reevaluated and reclassified as rachipagus. Two had extensive vertebral fusion and 18 were united in the dorsal midline, all with a meningocele, vertebral anomaly, and/or bony or neural connection. A theory of the embryologic origin of these twins is proposed: two embryonic discs located on diametrically opposing aspects of a single amniotic cavity becoming united in the area of the closing neural folds. This again raises the controversy concerning "fission or fusion" in conjoined twins. PMID- 8711622 TI - Atypical aortic coarctation and innominate artery stenosis associated with clubfoot and lower leg ischemia in an infant. AB - We report findings in a male infant born with talipes equinovarus and ischemic necrosis of the left lower leg and foot associated with atypical coarctation of the ascending aorta and stenosis and hypoplasia of the proximal innominate artery. We hypothesize that a vasculopathy of unknown etiology occurred in utero that resulted in ascending aortic arch coarctation and innominate artery stenosis. The presence of a thrombus in the coarcted segment suggests that one or more emboli could have been responsible for vascular compromise and subsequent clubfoot deformity and lower leg ischemia. Alternatively, a vasculopathy similar to that hypothesized as occurring in the aorta might also have occurred in the left anterior tibial artery, causing structural deformity and soft tissue ischemia of the left lower leg and foot. We believe that this is the first report of this association of findings. PMID- 8711623 TI - [Results of early treatment can now be proven. Interview by Klaus Reinhardt]. PMID- 8711624 TI - [Clinical procedures in brain infarct and differential diagnosis]. AB - The outcome of stroke improves significantly, when recognized and treated within 3 hours and in certain cases not later than 6 hours. In addition, secondary preventive measures potentially ameliorate the fate of the patient as well, especially when the cause of stroke is known. In this first article we outline the practical diagnosis and evaluation of stroke, both in the acute stage and thereafter. The differential diagnosis is also considered. PMID- 8711625 TI - [Epidemiology and risk factors in stroke]. AB - Epidemiology focuses on frequency and distribution of diseases and their relevant risk factors. Epidemiologic data are the basis for development of therapeutic strategies, evaluation of prevention and rehabilitation. Stroke in terms of epidemiology includes ischemic brain infarction as well as intracerebral and subarachnoidal hemorrhage. Stroke is most prominent in the elderly with an annual incidence of about 3%. Stroke prevalence varies between 5 and 8%. The 1-year case fatality is about 42%. The mortality rate varies considerably between 20 and 80 per 100 000 in comparable populations in Europe. Hypertension, smoking and atrial fibrillation are the most prominent risk factors; diabetes and the elevated serum cholesterol also contribute. PMID- 8711626 TI - [Clinical symptoms and stroke syndrome]. AB - A careful clinical examination usually allows a precise topographic stroke diagnosis. This article reviews the main symptoms and signs and the vascular syndromes of stroke. Vascular territories of the brain are described and illustrated with two figures. PMID- 8711627 TI - [Cerebrovascular ultrasound diagnosis]. AB - Introduction of color-coded duplex sonography has further increased diagnostic confidence of neurovascular ultrasound. Thus, reliable and noninvasive preoperative assessment of patients with carotid artery stenoses becomes feasible, preventing potential adverse effects of cerebral angiography. Moreover, reliable diagnosis of subtotal stenoses and occlusions of the carotid arteries is possible. Transcranial ultrasonography provides reliable assessment of cross-flow through the circle of Willis, and stenoses, occlusions and vasospasm of the major basal cerebral arteries. Transcranial color-coded duplex sonography identifies intracranial hemorrhage, cerebrovascular malformations and hydrocephalus but is inferior to neuroradiological techniques. It is likely that power Doppler sonography and transpulmonary contrast agents will provide an additional increase in diagnostic confidence of neurovascular ultrasonography. In the future, cerebral microemboli may become a marker for embolic activity of occlusive carotid artery and cardiac disease. PMID- 8711628 TI - [Neuroradiological studies and findings in stroke]. AB - This overview is about today's radiologic diagnostic possibilities in acute stroke. Despite many improvements in MR technology, CT is still the method of choice for most of these patients. Contrary to a long existing opinion, CT is a good diagnostic instrument even in the early phase of acute ischemic stroke. In combination with the new helical CT technique [CT angiography] all important questions regarding early therapeutic decisions can be answered. New MR perfusion techniques are just on the way to clinical application. Invasive angiography is only rarely indicated in acute ischemic stroke. The diagnosis of intracerebral hemorrhage is also mainly CT-based. MR and DSA are mandatory for further evaluation of the etiology of the hemorrhage. Nowadays, DSA is not longer mandatory for the diagnosis of cerebral venous sinus thrombosis. In some patients CT is already diagnostic; the combination of different MR and MRA techniques nearly always allows a definite diagnosis or exclusion of a venous thrombosis. PMID- 8711629 TI - [Cerebral artery dissection]. AB - The main symptom of arterial dissection is intense acute unilateral headache. The pain is commonly located around the eye, in the temple or the front with a carotid artery dissection [CAD] and in the posterior neck and occiput with a vertebral artery dissection [VAD]. Transient or persistent cerebral ischemic symptoms are similarly frequent but usually occur later in the time course. Horner's syndrome indicating a lesion of perivascular sympathetic fibres represents the third leading symptom and occurs in more than one third of the patients. Compression of local structures such as lower nerve or radicular palsies is rare. This constellation of symptoms in a young patient without vascular risk factors should rise suspicion of a dissection, in particular, if there is a preceding 'trivial' trauma. Characteristic features on Doppler/duplex sonography provide the diagnosis of dissection in almost all CAD and the majority of VAD. MRI demonstrating the mural hematoma allows reliable confirmation of the suspected diagnosis. Angiography is necessary only in selected cases, more often in VAD than in CAD. Brain infarction may be prevented, if premonitory symptoms, which occur in 60 to 80% of the patients, are recognized as such. Therefore, if there is clinical and sonographic suspicion of CAD or VAD, anticoagulation therapy with heparin should should be started before other imaging procedures finally prove the diagnosis. Because immediate anticoagulation may prevent cerebral embolism, this treatment strategy seems appropriate, although its efficacy has not been established by a controlled study. Anticoagulation should be continued until resolution of the dissection. PMID- 8711630 TI - [Cerebral sinus and venous thrombosis]. AB - Cerebral sinus and venous thrombosis account for less than 1% of all strokes, but diagnosis is often overlooked due to the variety of clinical syndromes and the variable clinical course. Underlying conditions do not significantly differ from those seen in extracerebral venous thrombosis. Conventional intra-arterial four vessel angiography was long the only diagnostic technique for confirming diagnosis, but nowadays magnetic resonance imaging and angiography are sufficient in many cases. Full-dose heparin treatment is the therapy of choice and is performed even if an hemorrhagic infarct already exists. If treatment is initiated early, the mortality rate of nonseptic cerebral sinus and venous thrombosis is less than 10%. According to general rules of postthrombotic treatment, patients should be treated with oral anticoagulants for a period of at least three months. PMID- 8711631 TI - [Vasculitis]. AB - Vasculitis, restricted to the central nervous system, may be an iodiopathic disorder or may be secondary to toxins, infections or neoplasm. Alternatively, it may be part of a systemic autoimmune disease. Temporal arteritis presents with headache, jay claudication, polymyalgia rheumatica and constitutional symptoms. Hemispheric strokes occur in 10%, especially in the territory supplied by the posterior or the middle cerebral arteries. Corticosteroids are the treatment of choice. Isolated angitis of the central nervous system is an idiopathic recurrent inflammatory disease of the small and medium-sized vessels, confirmed to the brain and the spinal cord. Persisting headaches, encephalopathy with personality changes, cognitive disorders and memory disturbances as well as multifocal neurologic deficits resulting from recurrent cerebral ischemia and cranial nerve involvement are the most frequent presently symptoms of the disease. Angiography of the intracranial vessels presents segmental stenoses or dilatations of the medium and small vessels; a leptomeningeal biopsy demonstrating vasculitis is the only reliable diagnostic method. Treatment is instituted with prednisone and cyclophosphamide. At least one year of therapy is necessary. The typical dermatological manifestations of Behcet's disease are aphthous stomatitis, genital ulcerations and ocular inflammation. Neurological involvement has been described in 30% of all patients with Behchet's disease. Sinus thrombosis presents with the signs of intracranial hypertension. A meningoencephalitis with headaches, disturbances of consciousness and focal neurologic symptoms is frequent; spinal cord involvement occurs. The treatment of choice for Behcet's disease consists of steroids in combination with chlorambucil or azathioprine. While polyarteritis nodosa and the Churg-Strauss syndrome most frequently involve the peripheral nervous system. CNS involvement is encountered in Wegener's granulomatosis. In lupus erythematosus and Sjorgren's syndrome, cerebral symptoms are rarely caused by a true vasculitis. In lupus erythematosus, nervous system involvement has been attributed to autoantibodies which interact directly with neurons [antineuronal antibodies] or indirectly lead to thrombotic vasculopathy by influencing the coagulation system [antiphospholipid antibodies]. PMID- 8711632 TI - [Intensive care in acute ischemia]. AB - Therapy of ischemic infarction has changed enormously during the last decade. This is due to an increase of pathophysiological knowledge on cerebrovascular disease, new diagnostic and intensive-care methods and success in thrombolytic therapy strategies; new neuroprotective drugs are on the way to be used on a routine base. The old fatalistic attitude towards treatment of stroke can nowadays not be justified. The success of stroke treatment is dependent on a fast and consequent therapeutic performing: patients with ischemic stroke who reach the hospital within six hours after onset of symptoms are candidates for thrombolysis. If certain inclusion criteria are fulfilled. Increasingly intracranial pressure is one of the major problems during the acute phase. Anti edema treatment follows a stepwise scheme. In some selected cases we perform hemicraniectomy. Besides the routine intensive-care monitoring, further monitoring is necessary by evoked potentials, brain pressure probes and Doppler ultrasound. PMID- 8711633 TI - [Therapy and prevention of cerebral ischemia]. AB - Two large randomized studies - NINDS and ECASS - have shown that fibrinolytics, when given early enough, reduce morbidity and mortality in acute stroke. The treatment has to be initiated within three hours after onset of symptoms. In middle cerebral artery stroke, fibrinolysis can be performed within six hours from onset, guided by computed tomography criteria. Thereafter, s.c. low molecular-weight heparin can still improve the fate of the patient. Currently, the potential benefit or harm of unfractionated heparin and aspirin is not yet known. However, large-scale studies using standard heparin, aspirin of low molecular-weight heparin are in progress with results expected soon. After the acute stage of stroke, risk factors have to be eliminated or modified, and anticoagulants, platelet inhibitors and, in selected cases, carotid endarterectomy are useful in preventing recurrent stroke or other vascular events. PMID- 8711634 TI - [Subarachnoid hemorrhage]. AB - The rupture of a cerebral aneurysm is the main cause of a spontaneous subarachnoid hemorrhage. Complications after the initial hemorrhage are recurrent bleeds, hydrocephalus and ischemia. Main symptom of a subarachnoid hemorrhage is the sudden, intense headache, often followed by loss of consciousness. The diagnosis is made by CT. If the CT is negative, lumbar puncture has to be performed, because in 5% of patients the blood may not be seen on the CT. If the diagnosis is confirmed, the patients has to be transferred as an emergency to a neurosurgical center. If his condition is satisfactory, the aim of the treatment is to occlude the ruptured aneurysm by microsurgical technique within the first three days after the hemorrhage. In patients with high surgical risk or bad general condition, the aneurysm can also be treated by the interventional neuroradiologist with endovascular techniques. PMID- 8711635 TI - [Intracerebral hemorrhages]. AB - Next to cerebral infarcts, spontaneous intracerebral hematomas are the second most frequent of cerebral insults. In recent years, the risk factors have been re evaluated. The prognosis with respect to morbidity and mortality is also more favorably estimated as compared to previous years. The prognosis is determined by the size of the hematoma, its location, and the degree of intraventricular hemorrhage. The various clinical syndromes can be differentiated on the origin of the hematoma. In general, cerebellar hematomas over 20 ml volume must be surgically removed. We recommend surgical removal of lobar and putaminal hemorrhages only when life saving measures are necessary. PMID- 8711636 TI - [Surgical therapy of intracerebral hemorrhages]. AB - Level of consciousness and focal neurological deficits following spontaneous intracerebral hemorrhage are a consequence of tissue destruction and compression. Surgical indications are given, if compression is the principal factor. Accepted indications are superficial cerebral hemorrhages of more than 50 to 100 ml and cerebellar hemorrhages of more than 20 to 30 ml. If basal ganglia hematomas should be surgically evacuated is still a matter of discussion. Arterial aneurysms and arteriovenous malformations associated with some spontaneous hematomas require special treatment strategies. PMID- 8711637 TI - Hepatotoxicity of antituberculosis drugs. PMID- 8711638 TI - Acute lung injury following lung resection: is one lung anaesthesia to blame? AB - Further examination of the parameters of oxidative stress, perioperative changes in the vasoregulatory mechanisms of the pulmonary circulation, and characterisation of the endothelial insult that probably occurs in all patients undergoing lung resection is necessary if the operative conditions under which lung surgery is carried out are to be optimised. Perhaps, then, more insight might be gained into how to improve preservation of lungs for transplantation and how to protect the lung from significant injury following resection. PMID- 8711639 TI - Pulmonary rehabilitation at home. PMID- 8711640 TI - In vivo effects of recombinant human DNase I on sputum in patients with cystic fibrosis. AB - BACKGROUND: Viscoelastic secretions in cystic fibrosis cause impaired mucus clearance and persistence of bacteria within the lung. The abnormal rheology is partly due to the presence of high molecular weight deoxyribonucleic acid (DNA). Recombinant human DNase I (rhDNase) has been shown to depolymerise DNA and thereby reduce the in vitro viscoelasticity of sputum in patients with cystic fibrosis. A phase II double blind placebo controlled study showed that rhDNase improved pulmonary function in patients with cystic fibrosis. The object of the present study was to evaluate the in vivo effects of rhDNase on sputum rheology and to determine whether these were correlated with changes in pulmonary function. METHODS: Patients were randomised to receive either placebo or rhDNase 2.5 mg twice daily for 10 days. Sputum samples were collected in sterile containers during screening and during treatment with the study drug. Pulmonary function and rheological analysis were the primary outcomes evaluated. Other parameters assessed were quantitative sputum bacteriology, sputum DNA concentration, and change in molecular mass of DNA polymers. RESULTS: The viscoelasticity of the sputum in untreated patients with cystic fibrosis was high and treatment with rhDNase reduced all the rheological parameters measured: dynamic storage modulus (a measure of elasticity), dynamic loss modulus (a measure of viscosity), and log complex modulus (a measure of mucus rigidity). The calculated cough clearance index was also improved following treatment with rhDNase. These rheological parameters showed a correlation with forced expiratory volume in one second (FEV1) which was improved by a mean (SE) of 13.3 (5.6)% on day 10 of treatment with rhDNase compared with a change of 0.2 (3.1)% in the placebo group. There was no change in bacterial colony counts or sputum DNA concentrations following treatment with rhDNase, but a small decrease in high molecular weight DNA was observed. CONCLUSIONS: Patients with cystic fibrosis treated with rhDNase show an improvement in rheological properties and pulmonary function, one of the mechanisms being a reduction in the proportion of high molecular weight DNA. PMID- 8711641 TI - Resting energy expenditure and oxygen cost of breathing in patients with cystic fibrosis. AB - BACKGROUND: Resting energy expenditure (REE) is often increased and may contribute towards energy imbalance in patients with cystic fibrosis. Several mechanisms may lead to increased REE including the gene defect, the effect of chronic infection, and abnormal pulmonary mechanics. Increased oxygen cost of breathing (OCB) has been demonstrated in patients with chronic obstructive pulmonary disease (COPD), but has not been the subject of extensive study in cystic fibrosis. METHODS: Ten clinically stable patients with cystic fibrosis and 10 healthy control subjects were studied. OCB was estimated using the dead space hyperventilation method. Mixed expired gas fractions were measured by online gas analysers and ventilation by a pneumotachograph. After measurement of resting ventilation and gas exchange, minute ventilation (VE) was stimulated by 6-10 1/min by the addition of a dead space and OCB calculated from the slope of the differences in oxygen uptake (VO2) and VE. REE and the non-respiratory component of REE were calculated from gas exchange data. To assess the repeatability of OCB all subjects had a further study performed one week later. RESULTS: The patients had lower weight, fat free mass (FFM), forced expiratory volume in one second (FEV1), forced vital capacity (FVC), and transfer factor for carbon monoxide (TLCO) than controls. Resting respiratory rate, VE, and oxygen uptake per kilogram of FFM (VO2/kg FFM) were higher in patients (20 (7), 10.4 (1.4) 1/min and 5.5 (0.8) ml/kg FFM/min) than in controls (13 (4), 7.0 (1.2), and 4.2 (0.5), respectively.) The error standard deviation for replicated measures of OCB was 0.5 ml O2/l VE in controls and 0.8 ml O2/l VE in patients with coefficients of variation of 24% in controls and 28% in patients. The mean OCB in patients was 2.9 (1.4) ml O2/l VE and 2.1 (0.7) ml O2/l VE in controls. OCB, expressed as ml/min (VO2resp) was 28.5 (11.7) in patients and 14.0 (3.6) in controls. REE was higher in patients (125.9 (14.0)% predicted) than in controls (99.0 (9.4)%). The estimated non-respiratory component of REE was 112.1 (14.9)% for patients and 93.0 (10.0)% for controls. CONCLUSIONS: In clinically stable patients with cystic fibrosis the OCB at rest is increased but is not the sole explanation for increased metabolic rate. This contrasts with the finding in COPD where the increase in REE is largely explained by increased OCB. This study also showed poor repeatability and OCB measurements similar to earlier studies, which indicates that the technique is not suitable for longitudinal studies. PMID- 8711642 TI - Risk factors for hepatotoxicity from antituberculosis drugs: a case-control study. AB - BACKGROUND: Several risk factors for the development of hepatotoxicity during short course antituberculosis therapy have been suggested. A case-control study was undertaken to assess the role of age, sex, disease extent, nutritional status, past history of liver disease, infection with hepatitis viruses, acetylator status, and high alcohol intake as risk factors in the development of hepatotoxicity in patients with pulmonary tuberculosis receiving antituberculosis treatment. METHODS: The cases comprised 86 consecutive patients who were diagnosed as having hepatitis induced by antituberculosis drugs and who were negative for any of the hepatitis markers (HAV-IgM, HBsAg, HBc-IgM, and anti HCV). The control group comprised 406 consecutive patients attending the chest clinic who completed antituberculosis treatment without developing hepatitis. The variables analysed were age, sex, body mass index (BMI), history of high alcohol intake, radiological extent of the disease, acetylator status, and serum proteins. RESULTS: The cases were older and their serum albumin levels were lower than in the control group. High alcohol intake was more common among the cases, they had more extensive disease radiologically, and the proportion of slow acetylators was higher. No differences were observed between the two groups in the other risk factors analysed. CONCLUSIONS: Of the various risk factors analysed, only advanced age, hypoalbuminaemia, high alcohol intake, slow acetylator phenotype, and extensive disease were risk factors for the development of hepatotoxicity. The risk of hepatitis in the presence of one or more of these risk factors may be increased. PMID- 8711643 TI - Comparison of characteristics of patients and treatment outcome for pulmonary non tuberculous mycobacterial infection and pulmonary tuberculosis. AB - BACKGROUND: Patients with non-tuberculous mycobacteria are usually started on conventional antituberculous triple therapy once acid fast bacilli are detected, before the exact type of mycobacteria has been identified. The ability to identify the characteristics of patients with tuberculous and non-tuberculous mycobacteria may be helpful in identifying before treatment those patients more likely to have non-tuberculous infection. METHODS: A retrospective study was conducted of all patients in one unit in whom non-tuberculous mycobacteria were identified in sputum or bronchoalveolar washings in the period 1987-93. The pattern of drug resistance was determined from laboratory records, and all case notes and chest radiographs were reviewed to identify the underlying disease and treatment outcome. All cases were compared with a matched control group of patients with culture positive Mycobacterium tuberculosis diagnosed during the same period. RESULTS: In the period studied there were 70 non-tuberculous and 221 tuberculous isolates. The non-tuberculous bacteria were typed as follows: M xenopi 23 (33%), M kansasii 19 (27%), M fortuitum 14 (20%), others 14 (20%). Of those with non-tuberculous mycobacteria, 83% were white subjects compared with 47% for tuberculosis. Patients with non-tuberculous mycobacteria were older than those with tuberculosis. Pre-existing lung disease or AIDS was present in 81% of patients with non-tuberculous mycobacteria and in 17% of patients with tuberculosis. Sensitivity to rifampicin and ethambutol was seen in 95% of M xenopi and 96% of M kansasii isolates. Relapse occurred in 60% of cases infected with M xenopi, 20% infected with M kansasii, and in 7% of cases with tuberculosis. CONCLUSIONS: In the population studied non-tuberculous mycobacteria occurred most frequently in elderly white subjects with pre-existing lung disease. If mycobacteria are detected in this group, consideration should be given to the possibility of non-tuberculous infection before embarking on treatment. A combination containing rifampicin and ethambutol is effective. The relapse rate for infection with M xenopi is high and prospective studies of the effect of the above combination of antituberculosis drugs are needed. PMID- 8711644 TI - Increasing incidence of tuberculosis in the young and the elderly in Scotland. AB - BACKGROUND: Since 1987 there has been an arrest in the previously established decline of tuberculosis notifications in Scotland. A study was undertaken to determine whether age contributed to this phenomenon. METHODS: Notifications of tuberculosis in Scotland were quantified by year and age group for the years 1981 92 from national statistics supplied by the Information and Statistics Division. Population data were obtained from the 1981 and 1991 national censuses. RESULTS: Age group analysis of pulmonary tuberculosis notifications showed that, in the 0 14 age group, incidence (per 10(5) population) decreased from 7.4 in 1981 to 2.6 in 1987, rising by an estimated 12.6% per annum to 3.7 in 1992. In the 65+ age group incidence declined from 30.1 in 1981 to 17.3 in 1988, and rose by an estimated 4.1% per annum to 22.2 in 1992. In the age groups 15-44 and 45-64 a continuous decrease in notification rate was seen over the period of the study. CONCLUSIONS: The plateauing of the incidence of tuberculosis in Scotland is associated with significant increases since 1987 of tuberculosis in the young and elderly. Contributions from ethnic minorities and those infected with HIV are negligible. An ageing population over the decade, with the highest tuberculosis rates seen in the older age group, may explain these findings. PMID- 8711645 TI - Interleukin 10 (IL-10) regulation of tumour necrosis factor alpha (TNF-alpha) from human alveolar macrophages and peripheral blood monocytes. AB - BACKGROUND: Regulation of the inflammatory response within the human lung is essential to prevent this important part of the normal host defence response becoming a pathological process. Tumour necrosis factor alpha (TNF-alpha) is a cytokine involved in the pathogenesis of shock and in granuloma formation, tissue necrosis, and fibrosis in many organ systems including the lung. Interleukin 10 (IL-10) has been proposed as having an inhibitory effect on the production of several inflammatory cytokines including TNF-alpha. METHODS: The effect of IL-10 administration on TNF-alpha production was explored in human alveolar macrophages and peripheral blood monocytes from matched individuals. The effects of IL-10 on TNF-alpha protein production were determined by sandwich enzyme linked immunosorbant assay (ELISA), whereas the TNF-alpha mRNA response was established by Northeren blotting using a TNF-alpha specific oligonucleotide probe. The protein synthesis inhibitors actinomycin D and cyclohexamide were utilised to monitor IL-10 effects on mRNA degradation and de novo protein synthesis, respectively. RESULTS: The lipopolysaccharide-mediated TNF-alpha production in alveolar macrophages was reduced from 3.508 (0.629) to 2.035 (0.385) ng/ml by 100 U/ml IL-10. Lipopolysaccharide-induced TNF-alpha production in peripheral blood monocytes was reduced from 2.035 (0.284) to 0.698 (0.167) ng/ml. TNF-alpha gene expression was also inhibited in both alveolar macrophages and peripheral blood monocytes; lipopolysaccharide-induced TNF-alpha mRNA was reduced by 47.8 (15.2)% and 83.1 (4.2)%, respectively, by IL-10. The IL-10 mediated suppression of TNF alpha mRNA was unaffected by addition of cyclohexamide, suggesting that de novo protein synthesis was not required for TNF-alpha inhibition. mRNA stability experiments indicated no acceleration in lipopolysaccharide-induced TNF-alpha mRNA degradation in response to IL-10. CONCLUSIONS: These findings suggest that IL-10 is a potent inhibitor of TNF-alpha expression and release from alveolar macrophages and peripheral blood monocytes, and thus it may have an important role in the cytokine network of the pulmonary immune response. PMID- 8711646 TI - Induction of interleukin 6 and interleukin 8 expression by Broncho-Vaxom (OM-85 BV) via C-Fos/serum responsive element. AB - BACKGROUND: Broncho-Vaxom (OM-85 BV) increases the resistance of the respiratory tract to bacterial infections by modulating host immune responses. The compound increases serum IgG levels but decreases IgE levels in patients suffering from chronic bronchitis or chronic obstructive pulmonary disease. It increases concentrations of gamma-interferon (IFN-gamma), IgA, and interleukin (IL)-2 in bronchoalveolar lavage fluid of patients with bronchitis. Treatment with OM-85 BV increases the number of T helper and natural killer cells. In this study the effects of OM-85 BV on transcription of cytokines is investigated in human lung fibroblasts. METHODS: Transcription and synthesis of IL-6 and IL-8 were assessed in cultured primary human lung fibroblasts using standard methods of Northern blot analysis for the level of mRNAs and enzyme linked immunosorbent assay for proteins. RESULTS: Broncho-Vaxom (OM-85 BV) at different concentrations induced transcription of IL-6 and IL-8. The effect of the drug on transcription of IL-6 and IL-8 genes correlated with secretion of the proteins into cell supernatants. OM-85 BV-dependent expression of the interleukin genes involved C-Fos/serum responsive element (C-Fos/SRE). CONCLUSIONS: The data suggest that the various immunopharmacological activities of OM-85 BV that have been described in clinical studies may be explained by its ability to induce expression of IL-6 and IL-8. PMID- 8711647 TI - Priming effect of platelet activating factor on leukotriene C4 from stimulated eosinophils of asthmatic patients. AB - BACKGROUND: Eosinophils from asthmatic patients are known to release greater amounts of leukotrienes than normal eosinophils when stimulated by the calcium ionophore A23187. The effect of platelet activating factor (PAF) in priming eosinophils was investigated. METHODS: Eosinophils were obtained from 18 asthmatic patients and 18 healthy donors. Cells separated by the Percoll gradients were incubated with PAF (C-18) for 30 minutes and then stimulated with the calcium ionophore A23187 (2.5 microM) for 15 minutes. The amount of leukotriene C4 (LTC4) in supernatants was measured using a combination of high pressure liquid chromatography and radioimmunoassay. RESULTS: The mean (SD) amount of LTC4 released by eosinophils from asthmatic patients upon stimulation with the calcium ionophore A23187 alone was 27.9 (9.9) ng/10(6) cells (n = 6). The amount of LTC4 released following stimulation with the calcium ionophore A23187 after pretreatment with PAF (1, 5, and 10 microM) was 57.2 (8.9), 75.1 (14.3), and 52.6 (10.7) ng/10(6) cells (n = 6), respectively. Trace amounts of LTC4 (0.9 (0.02) ng/10(6) cells, n = 6) were detected in the supernatant of the cells after stimulation by PAF alone (5 microM). The amount of LTC4 released upon stimulation by calcium ionophore A23187 alone in eosinophils from healthy donors was 10.3 (3.7) ng/10(6) cells (n = 4). The amounts of LTC4 released upon stimulation with calcium ionophore A23187 after pretreatment with PAF at concentrations of 1, 5, and 10 microM were 11.9 (3.5), 17.8 (5.6), and 12.7 (5.1) ng/10(6) cells (n = 4), respectively. Trace amounts of LTC4 (0.6 (0.02) ng/10(6) cells, n = 4) were detected in the supernatant of the cells upon stimulation with PAF alone (5 microM). The amounts of LTC4 released upon stimulation with calcium ionophore A23187 after pretreatment with lyso-PAF at concentrations of 1, 5, and 10 microM (n = 4 or 6) were 30.8 (5.2), 22.9 (5.1), and 27.3 (4.3) ng/10(6) cells (n = 6) from the eosinophils of asthmatic patients and 13.7 (3.3), 15.2 (4.9), and 14.7 (3.8) ng/10(6) cells (n = 4) from the eosinophils of healthy donors. CONCLUSIONS: The results indicated that PAF enhanced LTC4 formation by eosinophils obtained from asthmatic patients stimulated with the calcium ionophore A23187, but not those obtained from normal subjects. PMID- 8711648 TI - Evidence for opioid modulation and generation of prostaglandins in sulphur dioxide (SO)2-induced bronchoconstriction. AB - BACKGROUND: Inhalation of sulphur dioxide (SO2) provokes bronchoconstriction in asthmatic subjects. Cholinergic mechanisms contribute, but other mechanisms remain undefined. The effect of morphine, an opioid agonist, on the cholinergic component of SO2-induced bronchoconstriction was investigated, and the effect of indomethacin, a cyclooxygenase inhibitor, on SO2-induced bronchoconstriction and tachyphylaxis was studied. METHODS: In the first study 16 asthmatic subjects inhaled either ipratropium bromide or placebo 60 minutes before an SO2 challenge on days 1 and 2. On day 3 an SO2 challenge was performed immediately after intravenous morphine. In the second study 15 asthmatic subjects took either placebo or indomethacin for three days before each study day when two SO2 challenges were performed 30 minutes apart. The response was measured as the cumulative dose causing a 35% fall in specific airways conductance (sGaw; PDsGaw35). RESULTS: Ipratropium bromide significantly inhibited SO2 responsiveness, reducing PDsGaw35 by 0.89 (95% CI 0.46 to 1.31) doubling doses. This effect persisted after correction for bronchodilatation induced by ipratropium bromide. The effect of ipratropium bromide and morphine on SO2 responsiveness also correlated (r2 = 0.71). In the second study SO2 tachyphylaxis developed with PDsGaw35 on repeated testing, being reduced by 0.62 (95% CI 0.17 to 1.07) doubling doses. Indomethacin attenuated baseline SO2 responsiveness, increasing PDsGaw35 by 0.5 (95% CI 0.06 to 0.93) doubling doses. CONCLUSIONS: These results suggest that opioids modulate the cholinergic component of SO2 responsiveness and that cyclooxygenase products contribute to the immediate response to SO2. PMID- 8711649 TI - Prevalence and treatment of chronic airways obstruction in adults over the age of 45. AB - BACKGROUND: Chronic airways obstruction is common in adults and the elderly. To investigate the possibility that older adults with obstructive airways disease frequently do not receive appropriate treatment, the respiratory symptoms, medication use, and pulmonary function were studied in a random sample of white adults aged over 45 living in central Manchester, UK. METHODS: A postal questionnaire survey was performed on 783 men and women aged 45 years and over selected from GP lists by random number tables. Subjects completing the questionnaire were invited to attend for pulmonary function testing and methacholine challenge (Newcastle dosimeter method). RESULTS: The questionnaire response rate was 92.3% (723 subjects). The mean age of the population was 66.1 years and 57.2% were women; 29.2% were current smokers and 37.3% were ex-smokers. Asthma or bronchitis was reported by 30.0%. Two hundred and forty seven representative subjects attended for pulmonary function testing and spirometric evidence of chronic airways obstruction was found in 26.4%. Respiratory symptoms were reported by 76.6% of subjects with chronic airways obstruction; 55.0% had features which may predict potential improvement on treatment (increased non specific bronchial responsiveness or significant bronchodilator reversibility). However, only 55.4% of subjects with airways obstruction had received a diagnosis of asthma or chronic bronchitis and only 36.9% were using inhaled bronchodilators or steroids. CONCLUSIONS: Chronic airways obstruction is very common in adults in this inner city population, but is frequently overlooked. Most subjects with chronic airways obstruction are not receiving appropriate treatment. PMID- 8711650 TI - Asthma, airways responsiveness and air pollution in two contrasting districts of northern England. AB - BACKGROUND: To assess the possible magnitude of differences between normal populations an epidemiological investigation of asthma was conducted in two strongly contrasting districts of northern England--rural West Cumbria on the west coast and urban Newcastle upon Tyne on the east coast. METHODS: A cross sectional survey of randomly identified men aged 20-44 years was conducted in two phases: phase 1, a postal survey of respiratory symptoms and asthma medication in 3000 men from each district; and phase 2, a clinical assessment of 300 men from each district comprising investigator administered questionnaires, skin prick tests, spirometry, and methacholine challenge tests. RESULTS: The phase 1 (but not phase 2) study showed a small excess of "ever wheezed" in Newcastle (44% versus 40%), but neither phase showed differences between the two districts for recent wheeze or for other symptoms characteristic of asthma. There were also no differences with regard to diagnosed asthma, current asthma medication, spirometric parameters, or airways responsiveness. The prevalence of quantifiable airways responsiveness (PD20 < or = 6400 micrograms) was 27.7% in West Cumbria and 28.2% in Newcastle. Regression analyses showed that PD20 was negatively associated with atopy and positively with forced expiratory volume in one second (FEV1); that an association between PD20 and current smoking could be explained by diminished FEV1; and that PD20 was not related to geographical site of residence. CONCLUSIONS: Neither airways responsiveness nor the other parameters of diagnostic relevance to asthma varied much between the two study populations, despite the apparent environmental differences. The most obvious of these were the levels of outdoor air pollution attributable to vehicle exhaust emissions, the ambient levels of which were 2-10 fold greater in Newcastle. Our findings consequently shed some doubt over the role of such pollution in perceived recent increases in asthma prevalence. It is possible, however, that an air pollution effect in Newcastle has been balanced by asthmagenic effects of other agents in West Cumbria. PMID- 8711651 TI - Effect of altitude on spirometric parameters and the performance of peak flow meters. AB - BACKGROUND: Portable peak flow meters are used in clinical practice for measurement of peak expiratory flow (PEF) at many different altitudes throughout the world. Some PEF meters are affected by gas density. This study was undertaken to establish which type of meter is best for use above sea level and to determine changes in spirometric measurements at altitude. METHODS: The variable orifice mini-Wright peak flow meter was compared with the fixed orifice Micro Medical Microplus turbine microspirometer at sea level and at Everest Base Camp (5300 m). Fifty one members of the 1994 British Mount Everest Medical Expedition were studied (age range, 19-55). RESULTS: Mean forced vital capacity (FVC) fell by 5% and PEF rose by 25.5%. However, PEF recorded with the mini-Wright peak flow meter underestimated PEF by 31%, giving readings 6.6% below sea level values. FVC was lowest in the mornings and did not improve significantly with acclimatisation. Lower PEF values were observed on morning readings and were associated with higher acute mountain sickness scores, although the latter may reflect decreased effort in those with acute mountain sickness. There was no change in forced expiratory volume in one second (FEV1) at altitude when measured with the turbine microspirometer. CONCLUSIONS: The cause of the fall in FVC at 5300 m is unknown but may be attributed to changes in lung blood volume, interstitial lung oedema, or early airways closure. Variable orifice peak flow meters grossly underestimate PEF at altitude and fixed orifice devices are therefore preferable where accurate PEF measurements are required above sea level. PMID- 8711652 TI - Multiple pathogens in adult patients admitted with community-acquired pneumonia: a one year prospective study of 346 consecutive patients. AB - BACKGROUND: The purpose of this study was to assess the causes of community acquired pneumonia in adult patients admitted to hospital. METHODS: A prospective study was performed on 346 consecutive adult patients (54% men) of mean (SD) 49.3 (19.5) years (range 17-94) admitted to a university affiliated regional hospital in southern Israel with community-acquired pneumonia over a period of one year. Convalescent serum samples were obtained from 308 patients (89%). The aetiological diagnosis for community-acquired pneumonia was based on positive blood cultures and/or significant changes in antibody titres to Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis, respiratory viruses, Coxiella burnetii, Mycoplasma pneumoniae, Chlamydia pneumoniae, and Legionella sp. RESULTS: The aetiology of community-acquired pneumonia was identified in 279 patients (80.6%). The distribution of causal agents was as follows: S pneumoniae, 148 patients (42.8%); M pneumoniae, 101 (29.2%); C pneumoniae, 62 (17.9%); Legionella sp, 56 (16.2%); respiratory viruses, 35 (10.1%); C burnetii, 20 (5.8%); H influenzae 19 (5.5%); and other causes, 21 patients (6.0%). In patients above the age of 55 years C pneumoniae was the second most frequent aetiological agent (25.5%). In 133 patients (38.4%) more than one causal agent was found. CONCLUSIONS: The causal agents for community-acquired pneumonia in Israel are different from those described in other parts of the world. In many of the patients more than one causal agent was found. In all these patients treatment should include a macrolide antibiotic, at least in the first stage of their illness. PMID- 8711653 TI - Clinical picture of community-acquired Chlamydia pneumoniae pneumonia requiring hospital treatment: a comparison between chlamydial and pneumococcal pneumonia. AB - BACKGROUND: The importance of Chlamydia pneumoniae as a cause of pneumonia has remained controversial. The clinical picture of C pneumoniae and Streptococcus pneumoniae in patients admitted to hospital with community-acquired pneumonia was compared during a C pneumoniae epidemic in Finland. METHODS: Group I consisted of 24 patients in whom serological testing and bacterial culture indicated an association with C pneumoniae only, group II comprised nine patients with both C pneumoniae and S pneumoniae, and group III consisted of 13 patients with S pneumoniae only. RESULTS: The patients with C pneumoniae suffered from headache more frequently than the other patients (group I, 46%; group II, 11%; and group III, 15%) and had received antimicrobial treatment more often before admission to hospital (group I, 54%; groups II and III, 0%). The patients with C pneumoniae produced few good sputum samples and had suffered from respiratory symptoms longer than those with S pneumoniae (group I, 10 days; groups II and III, 4 days). C reactive protein values on admission were lowest in group I and highest in group II. The antimicrobial treatment provided in hospital covered C pneumoniae in 36% of cases in group I and 0% in group II, while S pneumoniae was covered in all patients. C pneumoniae and S pneumoniae together were associated with more severe disease and a longer stay in hospital. CONCLUSIONS: Pneumonia caused by C pneumoniae was milder but clinically resembled that caused by S pneumoniae, and required hospital treatment even among young patients. Mixed infections were common and should be taken into account when planning antimicrobial treatment for community-acquired pneumonia. Further studies with more patients are needed to evaluate the severity of C pneumoniae pneumonia. PMID- 8711654 TI - Effect of chronic continuous positive airway pressure (CPAP) therapy on upper airway size in patients with sleep apnoea/hypopnoea syndrome. AB - BACKGROUND: There is evidence to suggest that chronic continuous positive airway pressure (CPAP) therapy may produce reversible changes in upper airway morphology and function in patients with sleep apnoea/hypopnoea. This study was designed to examine the effect of chronic CPAP therapy on upper airway calibre. METHODS: Twenty four men with the sleep apnoea/hypopnoea syndrome (mean (SE) apnoea/hypopnoea index 37 (5)) underwent lateral cephalometry with measurement of posterior airway space performed before and at least three months after initiation of CPAP therapy. RESULTS: There was no weight change between the two assessments and mean CPAP use was 4.8 (0.4) hours per night. Posterior airway space (PAS) was measured in erect and supine postures. PAS supine increased with CPAP therapy from a mean (SE) of 11.8 (0.8) mm to 13.4 (0.8) mm, but PAS erect did not. Correlation of the change in PAS (dPAS) before and after CPAP therapy showed an increase with increasing CPAP compliance measured as machine run time both for dPAS supine (r = 0.68) and dPAS erect (r = 0.47). CONCLUSIONS: Patients with the sleep apnoea/hypopnoea syndrome regularly using CPAP for more than four hours per night all showed an increase in dPAS supine. The use of chronic CPAP increases PAS supine probably by a reduction in upper airway oedema, and the change in size is dependent on CPAP use. PMID- 8711655 TI - Surgical treatment of myasthenia gravis in two major Middle East teaching hospitals: factors influencing outcome. AB - BACKGROUND: The results of thymectomy on patients with generalised myasthenia gravis have been widely reported. However, there is no information on whether the experience of western countries can be generalised to the population of the Middle East. The purpose of this study was to evaluate the safety and efficacy of thymectomy in patients with myasthenia gravis in a Middle East patient population and to identify clinical and histopathological factors associated with improved long term outcome of surgery. METHODS: In a prospective study, sixty three patients (aged 1.5-51 years) were treated in two university teaching hospitals between 1984 and 1991 and followed up for a mean of four years. Close communication was established with neurologists to obtain early referral. Radical anterior mediastinal dissection through a median sternotomy was performed in all patients. The response was evaluated by modified Osserman's classification. RESULTS: Eighteen patients achieved complete remission and a further 39 improved, producing an overall response rate of 90.5%. Patients with milder disease (stage II) had a higher response rate (97%) than those with more advanced disease (78%). Patients operated on with less than three years of symptoms had a better outcome (94%) than those with longer duration of preoperative symptoms, especially in non thymomatous patients. Age and sex had no effect on the outcome. There was no effect on response rate if patients had hyperplastic or non-specific thymic histological findings, but patients with thymoma fared worse. CONCLUSIONS: These results are comparable with reports from the western world and represent the first prospective study from the Middle East. Thymectomy is indicated for all patients suffering from generalised myasthenia gravis soon after the diagnosis is made, regardless of age, stage, thymic pathology, and preoperative clinical status. PMID- 8711656 TI - Transtracheal aspiration using rigid bronchoscopy and a rigid needle for investigating mediastinal masses. AB - BACKGROUND: Use of the flexible needle via the fibreoptic bronchoscope to aspirate mediastinal nodes or masses has largely superseded the use of the rigid needle via the rigid bronchoscope. However, the yield at fibreoptic bronchoscopy is relatively low, although this improves with the use of a wider gauge needle. In this study the sensitivity and the safety of rigid needle sampling of the mediastinum in the diagnosis of lung cancer is evaluated. METHODS: Transtracheal needle aspiration (TTNA) was performed with the rigid bronchoscope and a rigid aspiration needle under general anaesthesia using a previous computed tomographic (CT) scan as a guide to the sample site. A cytopathologist immediately examined the specimens for adequacy and preliminary diagnosis, thus determining the number of aspirations. RESULTS: Twenty four patients were evaluated. The diagnostic sensitivity of TTNA was 88%. This led to a management decision in 21 patients. There were no false positives and no complications. CONCLUSIONS: TTNA using the rigid bronchoscope with CT scanning and a cytopathologist present is a sensitive and safe way of diagnosing lung cancer in patients with a mediastinal mass or enlarged mediastinal nodes. PMID- 8711657 TI - Angiotensin converting enzyme (ACE) inhibitor-induced cough and substance P. AB - BACKGROUND: Angiotensin converting enzyme (ACE) inhibitors cause coughing in 5 10% of patients, but the exact mechanisms of this effect are still unclear. In the airways ACE degrades substance P so the cough mechanism may be related to this peptide. METHODS: Nine patients who developed a cough and five patients who did not develop a cough when taking the ACE inhibitor enalapril (2.5 or 5.0 mg/day) for hypertension were enrolled in the study. No subjects had respiratory disease and the respiratory function of all subjects was normal. One month after stopping enalapril, inhalation of hypertonic saline (4%) was performed using an ultrasonic nebuliser for 15-30 minutes to induce sputum. The concentration of substance P in the sputum sample was measured by radioimmunoassay. In four of the nine cases with a cough enalapril was given again for 1-2 weeks and the concentration of substance P in the induced sputum was again measured. RESULTS: One month after stopping enalapril the mean (SE) concentration of substance P in the sputum of the group with a cough was 16.6 (3.0) fmol/ml, significantly higher than that in the subjects without a cough (0.9 (0.5) fmol/ml). All four subjects in the group with a cough who were given a repeat dose of enalapril developed a cough again, but the concentrations of substance P in the induced sputum while taking enalapril (17.9 (3.2) fmol/ml) were similar to the values whilst off enalapril (20.0 (2.5) fmol/ml). CONCLUSIONS: The mechanisms of ACE inhibitor induced coughing may involve substance P mediated airway priming. However, the final triggering of the ACE inhibitor-induced coughing is unlikely to be due to this peptide. PMID- 8711658 TI - Assessment of oxygen supplementation during air travel. AB - BACKGROUND: The aim of this study was to simulate an in flight environment at sea level with a fractional inspired concentration of oxygen (FiO2) of 0.15 to determine how much supplemental oxygen was needed to restore a subject's oxygen saturation (SaO2) to 90% or to the level previously attained when breathing room air (FiO2 of 0.21). METHODS: Three groups were selected with normal, obstructive, and restrictive lung function. Using a sealed body plethysmograph an environment with an FiO2 of 0.15 was created and mass spectrometry was used to monitor the FiO2. Supplemental oxygen was administered to the patient by nasal cannulae. SaO2 was continuously monitored and recorded at an FiO2 of 0.21, 0.15, and 0.15 + supplemental oxygen. RESULTS: When given 2 l/m of supplemental oxygen all patients in the 15% environment returned to a similar SaO2 value as that obtained using the 21% oxygen environment. One patient with airways obstruction needed 3 l/m of supplemental oxygen to raise his SaO2 above 90%. CONCLUSIONS: This technique, which simulates an aircraft environment, enables an accurate assessment to be made of supplemental oxygen requirements. PMID- 8711659 TI - Pulmonary capillary blood volume in patients with probable pulmonary Kaposi's sarcoma. AB - BACKGROUND: An increase in pulmonary capillary blood volume secondary to angiogenesis has been described in Kaposi's sarcoma. The value of the pulmonary capillary blood volume as an early marker of pulmonary Kaposi's sarcoma was evaluated. METHODS: In a prospective study 45 HIV positive patients (nine asymptomatic for Kaposi's sarcoma, 29 with cutaneous or mucocutaneous Kaposi's sarcoma, and seven with pulmonary Kaposi's sarcoma), underwent pulmonary function tests and determination of transfer capacity for carbon monoxide (TLCO) with its components, pulmonary capillary volume and membrane factor. RESULTS: Total lung capacity (TLC), TLCO, and its components were similar in the three groups. TLCO was normal in patients with pulmonary Kaposi's sarcoma and no changes in membrane factor or pulmonary capillary volume were observed. CONCLUSION: Pulmonary function tests and pulmonary capillary volume alone are not useful for identifying patients with pulmonary Kaposi's sarcoma. PMID- 8711660 TI - Exercise-induced asthma and cardiovascular fitness in asthmatic children. AB - BACKGROUND: The role of physical training in the management of children with exercise-induced asthma is controversial. A study was undertaken to determine whether a relationship could be found between the occurrence of exercise-induced asthma and the degree of cardiovascular fitness in asthmatic children. METHODS: Twenty eight children aged 6-13 with mild to moderate asthma and dyspnoea during or after physical exercise were tested. All patients had a basal forced expiratory volume in one second (FEV1) of > 80% predicted. Twelve patients were taking corticosteroid maintenance medication by inhalation and 16 were not. Two exercise tests were performed on a treadmill to assess peak oxygen consumption rate (VO2max) and the percentage decrease in FEV1 after exercise. RESULTS: There was no correlation between the VO2max and the percentage decrease in FEV1. Patients not taking steroids showed a greater fall in FEV1 than those receiving corticosteroid medication (mean fall in FEV1 28.7% versus 6.6%). Four of the 12 children treated with steroids and two of the 16 children not taking steroids had a level of cardiovascular fitness lower than the 5th percentile for healthy Dutch children. CONCLUSION: Normal cardiovascular fitness does not prevent exercise induced asthma. PMID- 8711661 TI - New developments in asbestos-related pleural disease. PMID- 8711662 TI - NO or no NO in asthma? AB - Nitric oxide (NO) plays a key role as a vasodilator, neurotransmitter, and inflammatory mediator in the airways and is produced in increased amounts in asthma. It may have beneficial effects on airways function as a bronchodilator and is the neurotransmitter of bronchodilator nerves in human airways. On the other hand, NO may have deleterious effects on the airways as a vasodilator by increasing plasma exudation, and may also amplify the asthmatic inflammatory response. Proinflammatory cytokines and oxidants increase the expression of an inducible form of NO synthase (NOS) in airway epithelial cells in asthma, and this may underlie the increased levels of NO found in exhaled air of asthmatic patients. Inducible NOS is inhibited by glucocorticoids, but selective inhibitors of this enzyme may have therapeutic potential in asthma. PMID- 8711663 TI - Pulmonary actinomycosis appearing as a "ball-in-hole" on chest radiography and bronchoscopy. AB - A 40 year old diabetic man with pulmonary actinomycosis was admitted to hospital with recurrent haemoptysis. The chest radiograph showed an air meniscus in the left upper lobe, a rare presentation of pulmonary actinomycosis. Bronchoscopic examination revealed a mass in a cavity which has never been reported previously. He underwent lobectomy and the surgical specimen revealed sulphur granules, the typical pathological finding of actinomycosis, without evidence of fungal or mycobacterial infection. PMID- 8711664 TI - Primary malignant melanoma of the bronchus. AB - A rare case of primary malignant melanoma of the bronchus is described. Before considering this diagnosis, metastasis from an occult primary must be convincingly excluded and the tumour should conform to certain guidelines. This patient presented with a solitary lung tumour for which she underwent left lower lobectomy and continues to be tumour free 54 months after surgery. Two possible mechanisms of aetiology are suggested. PMID- 8711665 TI - Respiratory failure due to tracheobronchomalacia. AB - A case is described of tracheobronchomegaly progressing to extensive tracheomalacia, complicated by episodic choking, recurrent pulmonary infections, and irreversible hypercapnic respiratory failure. A Y-shaped tracheobronchial stent was placed endoscopically to splint the trachea open, with excellent clinical and physiological improvement. New stent designs may provide long term palliation in selected cases of diffuse tracheal collapse or stenosis, and offer an alternative to surgical repair. PMID- 8711666 TI - Commentary: yet more to see down the bronchial tree? PMID- 8711667 TI - Bronchiectasis and homozygous (P1ZZ) alpha 1-antitrypsin deficiency. PMID- 8711668 TI - Pulmonary diseases and HIV infection. PMID- 8711669 TI - Domiciliary ventilation in chronic obstructive pulmonary disease: where are we? PMID- 8711670 TI - Genetic analysis of IgE. PMID- 8711671 TI - Humoral control of airway tone. PMID- 8711672 TI - Mechanisms of cigarette smoke induced increased airspace permeability. AB - BACKGROUND: Increased epithelial permeability of the airspaces occurs commonly in the lungs of cigarette smokers. It is likely to be important in augmenting the inflammatory response in the airspaces and hence may have a role in the pathogenesis of emphysema. It has previously been shown that intratracheal instillation of cigarette smoke condensate induces increased epithelial permeability in vivo in rats and in vitro in epithelial cell monolayers, associated with a disturbance in the lung antioxidant, glutathione (GSH). The aim of this study was to assess the role of neutrophils, GSH, and tumour necrosis factor (TNF) in the increased epithelial permeability following intratracheal instillation of cigarette smoke condensate. METHODS: Epithelial permeability of the airspaces was measured in rat lungs as the passage of intratracheally instilled 125-iodine labelled bovine serum albumin (BSA) into the blood. The permeability of a monolayer of human type II alveolar epithelial cells to 125I BSA was also measured. RESULTS: Cigarette smoke condensate produced a 59.7% increase in epithelial permeability over control values peaking six hours after instillation and returning to control values by 24 hours. Depletion of neutrophils and, to a lesser extent, macrophages by an intraperitoneal injection of antineutrophil antibody did not influence the increased epithelial permeability induced by cigarette smoke condensate. Although instillation of human recombinant TNF alpha produced an increase in epithelial permeability in the rat lung from 0.62 (0.61)% to 1.27 (0.08)%, only a trivial amount of TNF alpha was detected in bronchoalveolar lavage (BAL) fluid in vivo or in culture medium from BAL leucocytes obtained from animals treated with cigarette smoke condensate (94.9 (28.8) units/ml). Furthermore, antiTNF antibody did not abolish the increased epithelial permeability produced by cigarette smoke condensate. The role of GSH was assessed by measuring the changes in both the reduced (GSH) and oxidised form (GSSG) in lung tissue and in BAL fluid. One hour after instillation of cigarette smoke condensate there was a marked fall in the GSH content in the lung (from 809.8 (31.8) to 501.7 (40.5) nmol/g) in association with increased GSSG levels (from 89.8 (2.7) to 148.7 (48.8) nmol/g). This was followed by a return of GSH levels to control values, with a concomitant decrease in GSSG levels six hours after instillation. GSH levels in BAL fluid fell dramatically following cigarette smoke condensate (from 2.56 (0.30) to 0.31 (0.21) nmol/ml) and this fall was sustained up to six hours after instillation of cigarette smoke condensate. CONCLUSIONS: These studies suggest that neutrophils and TNF do not have a major role in the increased epithelial permeability induced by cigarette smoke condensate. However, the data support a role for the depletion of the antioxidant glutathione in the increased epithelial permeability caused by cigarette smoke condensate. PMID- 8711673 TI - Increase in non-specific bronchial hyperresponsiveness as an early marker of bronchial response to occupational agents during specific inhalation challenges. AB - BACKGROUND: Specific bronchial reactivity to occupational agents may decline after exposure in the workplace ceases leading to falsely negative specific inhalation challenges. A study was carried out to assess prospectively whether increases in nonspecific bronchial hyperresponsiveness could be useful in detecting the bronchial response to occupational agents during specific inhalation challenges. METHODS: Specific inhalation challenges were performed in 66 subjects with possible occupational asthma due to various agents. After a control day the subjects were challenged with the suspected agent for up to two hours on the first test day. Those subjects who did not show an asthmatic reaction were rechallenged on the next day for 2-3 hours. The provocative concentration of histamine causing a 20% fall (PC20) in the forced expiratory volume in one second (FEV1) was assessed at the end of the control day as well as six hours after each challenge that did not cause a > or = 20% fall in FEV1. The subjects who had a significant (> or = 3.1-fold) reduction in PC20 value at the end of the second challenge day were requested to perform additional specific inhalation challenges. RESULTS: The first test day elicited an asthmatic reaction in 25 subjects. Of the other 41 subjects five (12%, 95% confidence interval (CI) 4% to 26%) exhibited a > or = 3.1-fold fall in the PC20 value after the inhalation challenge and developed an asthmatic reaction during the second (n = 3) or third (n = 2) challenge exposure. The offending agents included persulphate (n = 1), wood dust (n = 2), isocyanate (n = 1), or amoxycillin (n = 1). These five subjects had left their workplace for a longer period (mean (SD) 21 (14) months) than those who reacted after the first specific inhalation challenge (8 (11) months). CONCLUSIONS: The increase in non-specific bronchial hyperresponsiveness after a specific inhalation challenge can be an early and sensitive marker of bronchial response to occupational agents, especially in subjects removed from workplace exposure for a long time. Non-specific bronchial hyperresponsiveness should be systematically assessed after specific inhalation challenges in the absence of changes in airway calibre. PMID- 8711674 TI - Pulmonary effects of short term selenium deficiency. AB - BACKGROUND: Selenium dependent glutathione peroxidase (GPx) reduces hydrogen peroxide (H2O2) and organic hydrogen peroxides in both normal and pathological states. Chronic dietary deficiency of selenium results in a gradual decrease in GPx and altered response to environmental stress. However, glutathione-S transferase (GST) isozymes may increase and compensate for chronic GPx deficiency. The pattern of antioxidant enzyme activity and immunolocalisation of various enzymes in rat lung has not been described in short term (< 3 weeks) acute selenium deficiency. METHODS: The time course of GPx depletion from rat lung (measured every five days in subgroups of rats) during acute dietary selenium deficiency was evaluated. After 20 days of depletion, enzyme activity of lung GPx, catalase, superoxide dismutase (SOD), glutathione reductase (GR), glucose-6-phosphodiesterase (G-6-PD), and GST were determined. Immunohistochemical localisation of GPx and SOD was also performed. The response to lethal hyperoxia (> 95%) in control and selenium deficient rats was then established. RESULTS: At 20 days, lung GPx activity in the rats fed a selenium deficient diet was one third less than in control animals who received a normal diet, while changes in blood enzymes between control and deficient animals were similar. Other lung enzyme activities remained normal with the exception of cyanide inhibited SOD activity measured in selenium deficient rat lungs which declined to approximately 50% of normal. Immunohistochemical localisation of GPx showed a generalised loss of the enzyme throughout the lung parenchyma with some possible sparing of activity in epithelial cells of the bronchioles. When exposed to lethal hyperoxia, selenium deficient animals were more susceptible than control rats. CONCLUSIONS: This is the earliest time at which dietary selenium deficiency has been shown to produce moderate loss of GPx activity. This change in activity was associated with increased susceptibility to pulmonary oxidant stress. However, the role of decreased SOD activity (presumed to represent copper, zinc SOD), although unexpected, may have been a major contributor to increased damage from hyperoxia. These results emphasise the complex potential interaction of elemental deficiency with the natural antioxidant response to lethal hyperoxia. PMID- 8711675 TI - Development of OASYS-2: a system for the analysis of serial measurement of peak expiratory flow in workers with suspected occupational asthma. AB - BACKGROUND: Serial peak expiratory flow (PEF) measurement is usually the most appropriate first step in the confirmation of occupational asthma. Visual assessment of the plotted record is more sensitive and specific than statistical methods so far reported. The use of visual analysis is limited by lack of widespread expertise in the methods. A computer assisted diagnostic aid (OASYS-2) has been developed which is based on a scoring system developed from visual analysis. This removes the requirement for an experienced interpreter and should lead to the more widespread use of the technique. METHODS: PEF records were collected from workers attending an occupational lung disease clinic for investigation of suspected occupational asthma and from workers participating in a study of respiratory symptoms in a postal sorting office. PEF records were divided into two development sets and two gold standard sets. The latter consisted of records from workers in which a final diagnosis had been reached by a method other than PEF recording. An experienced observer scored individual work and rest periods for the two development set PEF records; linear discriminant analysis was used to compare measurements taken from development set 1 records with visual scores. Two equations were produced which allowed prediction of scores for individual work or rest periods. The development set 2 was used to determine how these scores should be used to produce a whole record score. The first gold standard set was used to determine the whole record score which best separated those with and without occupational asthma. The second set determined the sensitivity and specificity of the chosen score. RESULTS: Two hundred and sixty eight PEF records were collected from 169 workers and divided into two development sets (81 and 60 records) and two gold standard sets (60 and 67 records). Linear discriminant analysis produced equations predicting the score for work periods incorporating five indices of PEF change and one for rest periods using seven indices. These equations correctly predicted the score for development set 1 work and rest periods on 61% of occasions (kappa = 0.47). The whole record score for development set 2 records, after weighting for definite or definitely no occupational effect, correlated with the visual score (correlation coefficient 0.86). Comparison with gold standard set 1 identified a cut off which proved to have a sensitivity of 75% and a specificity of 94% for an independent diagnosis of occupational asthma when applied to gold standard set 2. CONCLUSIONS: These results suggest that the sensitivity and specificity of analysing PEF records for occupational asthma using OASYS-2 approaches that of visual analysis, but it should be absolutely reproducible. The performance of OASYS-2 is more specific and approaches the sensitivity of other statistical methods of analysis. The evaluation of a large number of PEF records from workers exposed to different sensitising agents suggests that these results should be robust and should be repeatable in clinical practice. PMID- 8711676 TI - Impaired reflex responses to airway occlusion in the inspiratory muscles of asthmatic subjects. AB - BACKGROUND: Asthmatic subjects have an impaired capacity to activate the diaphragm during attempted maximal inspiratory efforts. Limb muscles require reflex facilitation to achieve maximal force. The reflex responses of inspiratory muscles to airway occlusion in asthmatic subjects were measured and compared with those in non-asthmatic control subjects. METHODS: Nine healthy asthmatic subjects breathed at a constant inspiratory flow through a low resistance valve. Random inspirations were transiently occluded for 250 ms. Surface electromyographic activity (EMG) was recorded over the scalene muscles, parasternal intercostal muscles, and the lateral chest wall overlying the diaphragm. The asthmatic subjects were studied with and without bronchoconstriction. Responses were compared with data from a matched group of 12 control subjects. RESULTS: Compared with the reflex responses to airway occlusion in control subjects, the duration of the initial short latency inhibition of inspiratory muscles was prolonged by 50% in asthmatic subjects and the size of the subsequent excitation was reduced by 30%. Bronchoconstriction reduced the time to the peak of the excitatory response in asthmatic subjects, although the values remained longer than in the control subjects. CONCLUSIONS: This study reveals impaired reflex excitation of inspiratory motoneurones in asthmatic subjects which could contribute to the reduced ability of these subjects to drive the diaphragm during maximal volitional efforts. PMID- 8711677 TI - Relationship between the inflammatory infiltrate in bronchial biopsy specimens and clinical severity of asthma in patients treated with inhaled steroids. AB - BACKGROUND: Current guidelines on the management of asthma advocate the use of anti-inflammatory treatment in all but mild disease. They define disease control in terms of clinical criteria such as lung function and symptoms. However, the relationship between the clinical control of the disease and inflammation of the airways is not clear. A cross sectional study was therefore undertaken to investigate the relationship between airways inflammation and measures of clinical control and bronchial hyperresponsiveness in asthmatic patients treated with inhaled steroids. METHODS: Twenty six atopic adults (19-45 years) with mild to moderate asthma (baseline forced expiratory volume in one second (FEV1) > or = 50% predicted, concentration of histamine causing a 20% fall in FEV1 (PC20) 0.02 7.6 mg/ml) on regular treatment with inhaled steroids entered the study. Diary card recordings during the two weeks before a methacholine challenge test and bronchoscopic examination were used to determine peak flow variability, symptom scores, and use of beta 2 agonists. Biopsy specimens were taken by fibreoptic bronchoscopy from the carina of the right lower and middle lobes, and from the main carina. Immunohistochemical staining was performed on cryostat sections with monoclonal antibodies against: eosinophil cationic protein (EG1, EG2), mast cell tryptase (AA1), CD45, CD22, CD3, CD4, CD8, CD25, and CD45RO. The number of positively stained cells in the lamina propria was counted twice by using an interactive display system. RESULTS: There were no differences in cell numbers between the three sites from which biopsy specimens were taken. The PC20 for methacholine was inversely related to the average number of total leucocytes, EG1+, and EG2+ cells, mast cells, CD8+, and CD45RO+ cells in the lamina propria. These relationships were similar for each of the biopsy sites. Symptom scores, beta 2 agonist usage, FEV1, and peak flow variability were not related to any of the cell counts. CONCLUSIONS: Infiltration of inflammatory cells in the lamina propria of the airways seems to persist in asthmatic outpatients despite regular treatment with inhaled steroids. The number of infiltrating leucocytes such as mast cells, (activated) eosinophils, CD8+, and CD45RO+ cells in bronchial biopsy specimens from these patients appears to be reflected by airway hyperresponsiveness to methacholine, but not by symptoms or lung function. These findings may have implications for the adjustment of anti-inflammatory treatment of patients with asthma. PMID- 8711678 TI - Interrelationships between diagnosed asthma, asthma-like symptoms, and abnormal airway behaviour in adolescence: the Odense Schoolchild Study. AB - BACKGROUND: The diagnosis of asthma is based on several characteristics including symptoms and suitable tests of airway lability. However, it is neither clear to what degree various tests and symptoms identify the same subjects, nor how these characteristics are best combined to diagnose asthma. The interrelationships between physician-diagnosed asthma, asthma-like symptoms, and abnormal airway function, as defined by four commonly used tests, have therefore been assessed. METHODS: A population based sample of 495 Danish schoolchildren aged 12-15 years, comprising 292 randomly selected subjects and 203 subjects considered at risk of having or developing asthma, was examined. Symptoms and background information were recorded by questionnaire. The test panel consisted of baseline forced expiratory volume in one second (FEV1%), provocation with treadmill exercise (EXE) and with inhaled methacholine (PD15), and monitoring of peak expiratory flow (PEF) twice daily for two weeks. RESULTS: The sensitivity for diagnosed asthma was highest for PD15 followed by PEF monitoring, whereas specificity for asthma or asthma-like symptoms was marginally higher with the other two tests. Most symptomatic subjects with any positive test were identified by PD15 alone (75%) or in combination with PEF monitoring (89%). PEF variability was more susceptible to treatment with inhaled steroids than the PD15 index. Although inter-test agreement was weak (kappa < 0.40 for all pairs), significant associations were found between PD15 and EXE, PEF and EXE, and FEV1% and PD15. CONCLUSIONS: The agreement between the four tests was weak. In particular, PEF variability and methacholine responsiveness seem to identify different varieties of airway pathophysiology. The combined use of methacholine provocation testing and PEF monitoring may be helpful as an epidemiological screening tool for asthma. PMID- 8711679 TI - Effect of maximum ventilation on abdominal muscle relaxation rate. AB - BACKGROUND: When the demand placed on the respiratory system is increased, the abdominal muscles become vigorously active to achieve expiration and facilitate subsequent inspiration. Abdominal muscle function could limit ventilatory capacity and a method to detect abdominal muscle fatigue would be of value. The maximum relaxation rate (MRR) of skeletal muscle has been used as an early index of the onset of the fatiguing process and precedes failure of force generation. The aim of this study was to measure MRR of abdominal muscles and to investigate whether it slows after maximum isocapnic ventilation (MIV). METHODS: Five normal subjects were studied. Each performed short sharp expiratory efforts against a 3 mm orifice before and immediately after a two minute MIV. Gastric pressure (PGA) was recorded and MRR (% pressure fall/10 ms) for each PGA trace was determined. RESULTS: Before MIV the mean (SD) maximum PGA MRR for the five subjects was 7.1 (0.8)% peak pressure fall/10 ms. Following MIV mean PGA MRR was decreased by 30% (range 25-35%), returning to control values within 5-10 minutes. CONCLUSIONS: The MRR of the abdominal muscles, measured from PGA, is numerically similar to that described for the diaphragm and other skeletal muscles. After two minutes of maximal isocapnic ventilation abdominal muscle MRR slows, indicating that these muscles are sufficiently heavily loaded to initiate the fatiguing process. PMID- 8711680 TI - Effect of L-arginine on renal blood flow in normal subjects and patients with hypoxic chronic obstructive pulmonary disease. AB - BACKGROUND: L-arginine is the precursor of endothelium derived nitric oxide (NO) and increasing the available substrate may increase the production of NO. This has been shown by local infusion in peripheral vascular beds but there are few studies of the effects during systemic infusion. Renal vasoconstriction is known to be important in the pathogenesis of cor pulmonale in patients with hypoxic chronic obstructive pulmonary disease (COPD). The effects of a systemic infusion of L-arginine on renal and aortic haemodynamics were therefore investigated in normal subjects and in patients with hypoxic COPD. METHODS: Ten normal volunteers were recruited from the research staff of King's College Hospital Six patients with COPD and hypoxia (arterial oxygen tension (PaO2) < 8.5 kPa) were recruited from the thoracic medicine outpatient clinic at King's College Hospital and five age and sex matched normal subjects were recruited from a group of normal subjects recruited from the database of the Department of Health Care for the Elderly as volunteers for medical research. There was no history of renal, cardiac, or hepatic disease. Baseline values of time averaged mean of the maximum instantaneous velocity (Tamx) and maximum velocity (Vmax) of blood flow in intrarenal arteries were obtained using colour flow Doppler ultrasound. Using the same technique, Vmax was obtained from the abdominal aorta just distal to the xiphisternum before and after infusion of L-arginine via a large peripheral vein (20 g in 100 ml sterile water over 30 minutes). RESULTS: In normal subjects L arginine increased blood velocity in the intrarenal vessels from a mean of 0.22 m/s to 0.26 m/s, an increase of 19.8%. There was no effect on arterial blood pressure, heart rate, or aortic blood velocity. L-arginine had no effect on intrarenal or aortic blood velocity in patients with hypoxic COPD. In age matched controls L-arginine increased blood velocity in the intrarenal vessels from a mean of 0.20 m/s to 0.26 m/s, an increase of 36.8%. There was no effect on arterial blood pressure, heart rate, or aortic blood velocity. CONCLUSIONS: L arginine, at the doses administered, increased renal blood flow, as assessed by renal arterial velocity. This effect was not seen in patients with hypoxic COPD but was present in age matched controls. This suggests that the abnormal renal vascular control seen in hypoxic patients with COPD may reflect a disturbance of the L-arginine/nitric oxide pathway. PMID- 8711681 TI - Impact of obstructive airways disease on quality of life in older adults. AB - BACKGROUND: Obstructive airways disease adversely affects quality of life, although relationships between quality of life and lung function have been shown to be weak. The relationships between the results of a quality of life questionnaire, spirometric tests, and methacholine bronchial challenge were investigated in a population sample of middle aged and elderly people. METHODS: A random population sample of the white population of Central Manchester, UK were contacted by post. Respondents were invited to undergo bronchial challenge with methacholine (Newcastle dosimeter method) and to complete the St George's Respiratory Questionnaire. This self-completed questionnaire quantifies quality of life as three component scores, with higher scores indicating greater impairment of quality of life. RESULTS: Two hundred and twenty seven subjects aged 45-86 years completed the St George's Questionnaire and performed spirometric tests; 190 completed the methacholine challenge. All quality of life scores were higher in subjects with a forced expiratory volume in one second (FEV1)/forced vital capacity (FVC) of < 65%, indicating impaired quality of life in subjects with airways obstruction. There was no relationship between quality of life and age. Multiple regression analysis showed independent relationships between quality of life scores and both baseline FEV1 and bronchial responsiveness. However, the amount of variation in quality of life attributable to variation in FEV1 or bronchial responsiveness was less than 10%. Subgroup analysis indicated that the quality of life score was independently associated with bronchial responsiveness and not FEV1 in subjects aged < 65 years, but with baseline FEV1 and not bronchial responsiveness in older subjects. CONCLUSIONS: Obstructive airways disease significantly impairs quality of life in adults. The reduction in quality of life in these patients is related to both baseline pulmonary function and non-specific bronchial responsiveness. The impact of airways obstruction on quality of life does not decrease with advancing age. PMID- 8711683 TI - Growth of acid fast L forms from the blood of patients with sarcoidosis. AB - BACKGROUND: Acid fast cell wall deficient forms (CWDF) of bacteria have been grown from blood, bronchial washings, and ocular anterior chamber fluid from patients with sarcoidosis. A monoclonal antibody raised against Mycobacterium tuberculosis whole cell antigen (H37RV) was used to characterise further CWDF grown from the blood of patients with sarcoidosis. METHODS: Blood from 20 patients with active sarcoidosis and from 20 controls was cultured using methods favourable for the growth of CWDF. Isolates were further characterised by indirect fluorescent antibody analysis using a monoclonal antibody highly reactive with M tuberculosis. RESULTS: CWDF were grown from the blood of 19 of 20 subjects with sarcoidosis. All isolates stained positively with the monoclonal antibody and with a modified Kinyoun stain. No organisms were grown from the blood of controls. CONCLUSIONS: These data demonstrate that CWDF can be grown from the blood of nearly all patients with active sarcoidosis. The results confirm that the organisms are mycobacterial in origin and are similar, if not identical, to M tuberculosis. Their role in the pathogenesis of sarcoidosis is unknown. PMID- 8711682 TI - Detection of the multidrug resistance marker P-glycoprotein by immunohistochemistry in malignant lung tumours. AB - BACKGROUND: The multidrug resistance marker P-glycoprotein (P-gp) was studied immunohistochemically in 78 primary malignant lung tumours. P-gp is a 170 kD transmembrane ATP dependent drug efflux pump which has been shown to be important in the resistance of some tumours to chemotherapy. Certain normal tissues express P-gp and tumours derived from these tissues are often insensitive to cytotoxic agents, showing raised P-gp levels innately or following chemotherapy or radiotherapy. METHODS: Samples from 78 patients undergoing surgery for primary malignant lung tumours were snap frozen and stained immunohistochemically using the monoclonal antibody C219 which reacts with a P-gp epitope. None of the study group had received chemotherapy or radiotherapy before surgery was performed. RESULTS: Twenty seven of the 78 lung tumours (34.6%) showed immunohistochemically detectable levels of P-gp which varied with tumour type; 17 of 54 squamous cell carcinomas (31.5%), seven of 15 adenocarcinomas (46.7%), and neither of two small cell carcinomas showing positive staining. In six of seven cases normal respiratory epithelium present showed the presence of P-gp. CONCLUSIONS: P-gp is immunohistochemically detectable in frozen tissue from a proportion of malignant lung tumours before exposure to radiotherapy or drugs associated with multidrug resistance. It may have a role in tumour resistance to cytotoxic drugs, but further clinical studies will be required to evaluate any correlation between P gp levels and response to treatment. PMID- 8711684 TI - Pulmonary function before and after anterior spinal surgery in adult idiopathic scoliosis. AB - BACKGROUND: Little is known about the long term effects of anterior spinal surgery on pulmonary function in adult patients with idiopathic scoliosis. A study was therefore undertaken of pulmonary function before and after anterior spinal surgery in this group of patients. METHODS: Fourteen patients (12 women) of mean age 26.5 years (range 17-50, 10 > or = 20 years) were studied. All 14 patients underwent thoracotomy and anterior arthrodesis, and five also underwent posterior arthrodesis. Scoliosis curves and lung volumes were measured preoperatively and at a mean follow up of 32 months (range 14-49) after surgery. RESULTS: At long term follow up after surgery the mean (SD) Cobb angle of the thoracic curve improved from 59 degrees (25 degrees) to 39 degrees (18 degrees), a correction of 31%. Despite this there was a fall in mean forced vital capacity (FVC) of 0.21 litres (95% confidence interval 0.04 to 0.38). In the patients who underwent anterior surgery without posterior surgery (n = 9) there was a fall in mean FVC of 0.31 litres (95% confidence interval 0.12 to 0.50); mean forced expiratory volume in one second and total lung capacity were also reduced and there was an increase in mean residual volume. CONCLUSIONS: Forced vital capacity is reduced at long term follow up in adult patients with idiopathic scoliosis who undergo anterior spinal surgery. The fall in FVC is small and is unlikely to be of clinical significance in patients with reasonable lung function in whom surgery is planned for prevention of curve progression or improvement of cosmetic appearance and pain. However, surgical intervention should not be undertaken in an attempt to improve pulmonary function. PMID- 8711685 TI - Particle size selection device for use with the Turbohaler. AB - BACKGROUND: Drug deposited within the upper airways of patients using dry powder inhalers does not contribute to the therapeutic effect but can result in unwanted local side effects and, when swallowed, may contribute to systemic effects. A chamber has been devised which uses the centrifugal force generated by the Turbohaler to remove large "non-respirable" particles with a view to minimising deposition in the upper airway. An in vitro study was performed to determine whether such a chamber could reduce the dose contained in coarse particles without having a significant effect on the "respirable dose". METHODS: The mouthpiece of a 200 micrograms Turbohaler was modified to allow a small volume chamber to be attached. The particle size distribution generated by the Turbohaler was assessed using a multi-stage liquid impinger with a flow rate of 60 l/min. The quantity of drug on each stage was quantified using an ultraviolet spectrophotometric technique. For each experiment 10 actuations were used to ensure adequate quantities of drug on each stage. Particles depositing on stages 3 + 4 have a diameter of < 6.8 microns and are arbitrarily referred to as the "respirable dose". The particle size distribution obtained using the Turbohaler (n = 10) was compared with that from the Turbohaler+ chamber (n = 11). RESULTS: The addition of the chamber resulted in the mean (SD) dose contained in larger "non-respirable" particles depositing on stages 1 + 2 being reduced from 52.2 (12.3) to 29.6 (6.9) micrograms per actuation. However, the chamber did not affect the "respirable" dose. The dose contained in particles with a diameter of < 6.8 microns from the standard Turbohaler was 91.1 (8.9) micrograms compared with 82.4 (18.6) micrograms when used with the chamber. CONCLUSIONS: These results indicate that it is possible to devise an effective particle size selection device for the Turbohaler. It may be possible to produce such devices for other dry powder inhalers, although the design would need to be tailored to each particular device. PMID- 8711686 TI - Radiographic features of staphylococcal pneumonia in adults and children. AB - BACKGROUND: Clinical and laboratory features do not accurately correlate with the cause of community acquired pneumonia. A study was performed to examine whether the radiographic features of staphylococcal pneumonia are sufficiently distinct to aid early diagnosis. METHODS: The chest radiographs of 34 patients (including eight children) with proven staphylococcal pneumonia were reviewed by two experienced observers using methods described previously. Features on presentation and follow up were noted. RESULTS: The most striking features were the presence of multilobar consolidation on presentation, cavitation, pneumatocoeles and spontaneous pneumothorax, together with a tendency to radiographic deterioration after admission in both adults and children. Some of these features are much less common with other causes of community acquired pneumonia. However, most of the cases did not have these classic features. CONCLUSIONS: The presence of certain radiographic features, including multilobar shadowing, cavitation, pneumatocoeles, and spontaneous pneumothorax, are seen with staphylococcal pneumonia in adults and children, but their absence does not exclude the diagnosis. PMID- 8711687 TI - Respiratory irritants encountered at work. PMID- 8711688 TI - Scorpion venoms: taking the sting out of lung disease. AB - Scorpion venoms contain specific toxins which block large conductance calcium activated potassium (BKCa) channels. Use of these toxins has shown that a significant proportion of the action of bronchodilators such as beta-agonists, theophylline, and nitric oxide occurs as a result of the opening of BKCa channels. Similarly, these toxins have shown that inhibitors of airway neurotransmission also operate via BKCa channels. Drugs that open BKCa channels may be alternative bronchodilators (possibly "airway selective" and with fewer side effects) as well as inhibitors of pathophysiological neurogenic influences in asthma, chronic coughing and sneezing, and chronic bronchitis. PMID- 8711689 TI - Spontaneous regression of a giant pulmonary bulla. AB - Gradual expansion of a lung bulla is common and may be associated with debilitating pulmonary symptoms. The aetiology of bulla expansion is unclear. Spontaneous regression, on the other hand, is rarely observed. The case is presented of a man in whom near complete spontaneous resolution of a giant pulmonary bulla occurred. This event was associated with dramatic improvement in the radiographic picture and pulmonary function. PMID- 8711690 TI - Sternocostoclavicular hyperostosis presenting with thoracic sinus formation. AB - Sternocostoclavicular hyperostosis (SCCH) is a condition which is well described in the Japanese literature but is rare in Western Europe. It is characterised by pain and swelling in the upper anterior part of the chest, which tends to be progressive. A patient is described with bilateral chronic discharging sinuses over the anterior ends of the clavicles in whom the diagnosis appeared to be one of SCCH. PMID- 8711691 TI - Lymphangitis carcinomatosa complicating primary malignant peritoneal mesothelioma. AB - A patient with malignant peritoneal mesothelioma and a diffuse pulmonary infiltrate is described. Computed tomographic scanning suggested lymphangitis carcinomatosa. This was confirmed on transbronchial biopsy to be due to metastatic mesothelioma. PMID- 8711692 TI - Pulmonary eosinophilia due to Trichomonas tenax. AB - A 39 year old woman was admitted with fever and an exacerbation of asthma. The chest radiograph showed extensive bilateral lung infiltrates, but transtracheal needle aspiration did not reveal any infectious organisms. The bronchoalveolar fluid lavage contained a large number of Trichomonas tenax and an increased percentage of eosinophils. Corticosteroids provided a dramatic response. Because of the relatively high frequency of colonisation of the human mouth by T tenax (about 20%), this organism should be considered in all cases of pulmonary eosinophilia. PMID- 8711693 TI - Lung disease induced by drug addiction. PMID- 8711694 TI - [Creutzfeldt-Jakob syndrome. A new variant in Great Britain]. PMID- 8711695 TI - [Stroke units--why now?]. PMID- 8711696 TI - [Tamoxifen--effects and side-effects]. PMID- 8711697 TI - [Surgery of mitral valve]. PMID- 8711698 TI - [Mitral valve plasty. A 12-year material]. AB - 46 patients underwent mitral valve repair at the Regional Hospital in Trondheim/St. Elisabeth Heart Clinic, during the period 1983-95. There were 20 female patients. The valve pathology was ischemic in 40%, rheumatic in 10% and other or unknown in 50%. The surgical technique included commissurotomy (n = 9), ring annuloplasty (n = 18), resection of posterior leaflet (n = 12), Kay annuloplasty (n = 10), shortening of chordae (n = 3) and other techniques (n = 2). Postoperative complications included wound infection (n = 1), pneumonia (n = 1), mediastinitis (n = 1), pleural effusion (n = 5), renal failure (n = 1) and multi-organ failure (n = 1). One patient required re-exploration for postoperative bleeding. Three (6.5%) patients died within 30 days of surgery due to low cardiac output (n = 2) and multi-organ failure (n = 1). Nine patients (21%) died later. At follow up, within one year of surgery, marked functional improvement was registered, with only one patient in NYHA class III and none in class IV. PMID- 8711699 TI - [Tamoxifen and endometrial cancer. A case report]. AB - Tamoxifen is an oestrogen antagonist with partial agonistic effects that is used extensively in the treatment of mammary carcinoma. We report a case of an 80 year old woman who was operated on for carcinoma of the mammary gland and developed endometrial carcinoma after ten years of tamoxifen treatment. Tamoxifen has been noted for its reportedly low incidence of side effects. Since 1985, however, several reports have associated tamoxifen with higher risk of endometrial carcinoma. We also review the relevant literature published during the last decade. PMID- 8711700 TI - [Acromion resection for shoulder impingement syndrome. Results after an open and a percutaneous surgical method]. AB - Anterior acromioplasty as described by Neer has been an effective procedure for shoulder impingement syndrome. Artroscopic acromioplasty is regarded as a technically demanding procedure. Since 1988 we have developed a closed percutaneous acromioplasty using motorized instruments. The purpose of this randomised clinical trial was to compare the results of open and closed percutaneous acromioplasty. We found a significant improvement in shoulder score, but no differences between the group treated with the open procedure and the group treated with the closed procedure. PMID- 8711701 TI - [Embolization of congenital pulmonary arteriovenous malformations]. AB - Patients with arteriovenous pulmonary malformations are at risk of developing secondary brain disease such as transient ischemic attacks, strokes or abscesses. Lethal haemothorax and haemoptysis also occur. 12 of 14 malformations in five patients were treated using a total of eight procedures. One patient experienced a transient unilateral hemiparesis, otherwise no complications occurred. None of the 43 deployed occlusion coils was lost through the fistulas. Complete occlusion was achieved in all lesions where coils could be placed in a stable position. One patient suffered a minor recurrence. The use of interlocking detachable coils (IDC) which can be retracted or repositioned prior to full deployment is recommended. PMID- 8711702 TI - [Lymphatic complications after lower limb vascular surgery]. AB - A series of five patients with lymph fistula and two with lymphocele is described. Lymphatic complications in the groin and thigh following infrainguinal arterial surgery may increase the risk of wound infection and prolong the stay in hospital. During operation for lymph fistula and lymphocele, precise identification of the site of lymph leakage during exploration of the wound is mandatory. This can be obtained by intradermal injection of Patent blue 2.5% medially and laterally on the dorsum of the foot approximately one hour before operation. Operation for lymph fistula should be performed as soon as the diagnosis has been verified, especially if a prosthetic graft has been used. A lymphocele should be treated conservatively unless the patient is suffering from localized pain or ischemia of the skin caused by pressure of the underlying swelling. PMID- 8711703 TI - [Reduced prescription of benzodiazepine preparations in general practice. Much to gain with little effort]. AB - Many patients experience side effects from prolonged use of benzodiazepines and analgesics containing codeine. Therefore 66 daily users of these drugs (average dose before intervention 34 DDD/month) were encouraged to take part in a cessation experiment. They were told that a large share of their symptoms which were the indication for benzodiazepines and analgesics might be side effects from this medication. They were also given a cessation plan. Over two years the prescriptions (measured in DDD) for this group were reduced by 57%, and 26% of the patients stopped taking these medicines. A strategy is proposed for cessation of use of benzodiazepines and codeine-containing analgesics in general practice. PMID- 8711704 TI - [Emergency treatment of stroke patients--what does the primary health care do?]. AB - Has new evidence concerning acute care for stroke patients changed medical practice in primary health care in Norway? What type of wards are currently used when stroke patients are admitted to hospital? These questions are discussed on the basis of data from telephone interviews with chief medical officers in a representative sample of 77 Norwegian local administrative areas (municipalities). 60% of the medical officers state that admission practices are more liberal now than five years ago. Less severe cases are admitted to hospital more often. 34% claim that there has been no change, and 1% report that the practice is stricter now than before. According to our informants, routines for transport, managing TIA and managing nursing home patients with acute stroke vary. 26% of all stroke patients have access to stroke units. In most cases (59%) an ordinary medical ward is the only option. PMID- 8711705 TI - [Rehabilitation of stroke patients in counties and municipalities. Status, development and aspects of cooperation]. AB - This article describes rehabilitation services for stroke patients in Norwegian health care system as of November 1994. A telephone interview was carried out with chief medical officers in a representative sample of 77 primary health care districts. 47% of the informants reported that a 60-year-old patient with severe hemiplegia would receive rehabilitation in primary health care, 35% reported that this would take place in a hospital. The most frequent single offer for the case in hand would be a nursing home (35%). According to the medical officers, co operation between primary and secondary health care on long-term rehabilitation of stroke patients is limited. Only 25% of our informants consider it useful. 67% claim that the rehabilitation provided at the secondary level is unsatisfactory. A review of plans currently prepared by the county health care authority shows that if they are implemented, stroke unit and rehabilitation wards will be the main components of future activities at this level of health care in Norway. PMID- 8711706 TI - [Improved information on drugs to patients. A project with a pharmacist in the therapeutic team at a cardiology department]. AB - Patients with cardiac disease knew more about medicaments after receiving medicament-related counselling by a pharmacist on discharge from hospital. This was demonstrated by a questionnaire answered by 49 patients whose answers were compared with the results of a study conducted in 1991, when patients did not receive information from a pharmacist in hospital. More patients were pleased with the information, and most of them were satisfied with the consultation with the pharmacist. Lack of time was stated to be the main reason why other hospital health care professionals often provided insufficient information on medicaments to patients. It was discovered that physicians did not record a proper history of the medication. This affected management of the therapy and the information given to patients. This investigation shows that a pharmacist can make an important contribution to the care of patients. Health care professionals should begin to discuss with the health authorities how best to organize the information patients receive on medicaments. PMID- 8711707 TI - [High-dose therapy with cytostatic agents as primary treatment of advanced breast cancer]. AB - Thousands of women with breast cancer have received high dose chemotherapy prior to the results from controlled clinical trials being known. As one of these patients the author reviews and discusses the results of the first randomised study from South Africa. High dose therapy with autologous stem cell support was compared with conventional chemotherapy in 90 young women with metastatic aggressive breast cancer. Though survival was short in both groups the disease free survival was doubled in the high dose group. A significant increase was found in response rate, duration of response and survival. Data from America show the cost effectiveness of this treatment to be comparable to that of other life saving therapies. A comparison is made with the absolute and relative survival benefit of simvastatin treatment. A Norwegian White Paper on high dose therapy does not include advanced breast cancer in the planned trial protocols. It is argued that future health planning should give high priority to the treatment of advanced breast cancer in young women. PMID- 8711708 TI - [Towards a new policy for preventive work?]. PMID- 8711709 TI - [Movement and group discussions]. PMID- 8711710 TI - [200 years of smallpox vaccination]. PMID- 8711711 TI - [Assessment by the Norwegian Patient Insurance. Treatment of "ulnar Bennett's fracture"]. PMID- 8711712 TI - [Medication and mental disease]. PMID- 8711713 TI - [Head physicians should prioritize clinical research]. PMID- 8711714 TI - [Heart diseases in foals. A literature review exemplified by 2 case reports]. AB - A review of the congenital and acquired heart diseases of foals is given on the basis of two patients. A 3-month-old foal with a history of collapse after exercise had a systolic murmur on all heart valves on both sides. Necropsy revealed endocarditis ulcerosa of the left atrioventricular valves. A 6-week-old foal with systolic murmur on the left atrioventricular valves and on the aorta showed, on ultrasonography, signs of endocarditis of the atrioventricular valves. This foal recovered after a long course of antibiotics. PMID- 8711715 TI - [Pioneers: veterinarians from earlier times (16). Peter Christian Abildgaard]. PMID- 8711716 TI - [Developments in cattle and swine health care and the existence of special outlook groups]. PMID- 8711717 TI - [Injury claim in the administration of clenbuterol-containing drugs]. PMID- 8711718 TI - [Improvement in fertility results in cattle]. PMID- 8711719 TI - [FIDIN theme afternoon on drugs and animal welfare. Welfare or non-existence?]. PMID- 8711720 TI - [Tumor lysis syndrome in a dog]. AB - The acute tumour lysis syndrome in humans as well as in dogs is caused by the acute lysis of tumour cells following chemotherapy. The release of intracellular products and their metabolites result in hyperkalemia, hyperphosphatemia, and hyperuricemia. The precipitation of uric acid and calciumphosphate in the kidneys can lead to hypocalcemia and acute renal failure which leads to fatal azotemia and arrhythmia's due to electrolyte disturbances. In this article the occurrence of the acute tumour lysis syndrome in a dog with malignant lymphoma is described. Suggestions to prevent the tumour lysis syndrome are made. PMID- 8711721 TI - [On communication]. PMID- 8711722 TI - [Toxicity of Liberty]. PMID- 8711723 TI - [Chronic pharyngitis in the horse]. PMID- 8711724 TI - [Veterinary antibiotic policy: responsible and effective? A viewpoint from industry]. PMID- 8711726 TI - Risk Assessment Issues for Sensitive Human Populations. Conference proceedings. Dayton, Ohio, 25-27 April 1995. PMID- 8711725 TI - More or less devious paths to the heart. On the early history of surgery of the thorax and its organs. PMID- 8711727 TI - Clinically relevant EEG studies and psychophysiological findings: possible neural mechanisms for multiple chemical sensitivity. AB - This paper addresses the evidence for the face, construct, and criterion-related validity of the olfactory-limbic/neural sensitization model for multiple chemical sensitivity (MCS). MCS is a poorly-understood, controversial condition in which low levels of environmental chemicals are reported to trigger disabling levels of illness in certain individuals. Neural sensitization processes could generate an endogenous amplification of responsivity to exogenous substances, thereby providing a plausible explanation for the apparent lack of a classical toxicological dose-response relationship in MCS. Convergent data from both survey and psychophysiological studies of MCS patients and of persons from the community without MCS, but who report elevated frequency of illness from chemical odors (cacosmics), support the involvement of the limbic system and the sensitizability of cacosmics, as predicted by the model. Recent studies show that cacosmics do sensitize their heart rate, blood pressure, and plasma beta-endorphin responses to repeated exposures to a novel laboratory procedure involving dietary manipulations over time. Cacosmia may represent a pathological form of neural plasticity. Taken together, the model and the available evidence suggest the need for more intensive investigation of MCS from the standpoint of possible neurobiological mechanisms affecting cognitive, emotional, and somatic functions. PMID- 8711728 TI - Potential animal model of multiple chemical sensitivity with cholinergic supersensitivity. AB - Multiple Chemical Sensitivity (MCS) is a clinical phenomenon in which individuals, after acute or intermittent exposure to one or more chemicals, commonly organophosphate pesticides (OPs), become overly sensitive to a wide variety of chemically-unrelated compounds, which can include ethanol, caffeine and other psychotropic drugs. The Flinders Sensitive Line (FSL) rats were selectively bred to be more sensitive to the OP diisopropylfluorophosphate (DFP) compared to their control counterparts, the Flinders Resistant Line (FRL) rats. The present paper will summarize evidence which indicates that the FSL rats exhibit certain similarities to individuals with MCS. In addition to their greater sensitivity to DFP, the FSL rats are more sensitive to nicotine and the muscarinic agonists arecoline and oxotremorine, suggesting that the number of cholinergic receptors may be increased, a conclusion now supported by biochemical evidence. The FSL rats have also been found to exhibit enhanced responses to a variety of other drugs, including the serotonin agonists m-chlorophenylpiperazine and 8-OH-DPAT, the dopamine antagonist raclopride, the benzodiazepine diazepam, and ethanol. MCS patients report enhanced responses to many of these drugs, indicating some parallels between FSL rats and MCS patients. The FSL rats also exhibit reduced activity and appetite and increased REM sleep relative to their FRL controls. Because these behavioral features and the enhanced cholinergic responses are also observed in human depressives, the FSL rats have been proposed as a genetic animal model of depression. It has also been reported that MCS patients have a greater incidence of depression, both before and after onset of their chemical sensitivities, so cholinergic supersensitivity may be a state predisposing individuals to depressive disorders and/or MCS. Further exploration of the commonalities and differences between MCS patients, human depressives, and FSL rats will help to elucidate the mechanisms underlying MCS and could lead to diagnostic approaches and treatments beneficial to MCS patients. PMID- 8711729 TI - Proposed animal neurosensitization model for multiple chemical sensitivity in studies with formalin. AB - A potentially promising line of animal research relevant to multiple chemical sensitivity (MCS) is that of sensitization in the central nervous system (CNS), particularly limbic pathways in the brain. Sensitization is the progressive and enduring enhancement in behavioral and neurochemical responses that occurs after repeated exposure to psychostimulants or environmental stressors. Since the onset and progression of sensitization has many parallels with that of MCS, it has been proposed that MCS may be initiated through a mechanism similar to the sensitization of CNS components occurring in the rodent. To test this hypothesis, female Sprague-Dawley rats were exposed to formalin vapors (FORM, 11 ppm) or water vapor (control) 1 h/day for 7 days. The next day, a saline injection was given followed by a cocaine injection (15 mg/kg, i.p.) 24 h later, and locomotor activity was monitored. Animals pretreated with repeated FORM inhalation demonstrated a significantly enhanced locomotor response to cocaine compared to controls, an indicator that specific limbic pathways may have been sensitized. At 4 weeks of withdrawal from FORM exposure, a subset of animals remained sensitized to a cocaine challenge. No differences were found between groups after a saline injection. In a second experiment, animals were screened prior to FORM or water exposure for their response to a novel situation, a measure believed to reflect an animal's general responsiveness to stimuli. Rats were divided into high responders (HR) or low responders (LR), based on their locomotion in a novel cage. Results from three behavioral tests demonstrated that HR and LR were differentially affected by exposure to FORM. In a passive avoidance test, HR and LR appeared to be different in their distribution of responses, while HR and LR responses in the FORM group were nearly identical. On the elevated plus maze test of anxiety, HR spent more time on the open arms than LR in both treatment groups, with significant differences between HR and LR in the FORM, but not water, treated group. On a hot plate test to measure nociceptive levels, no differences occurred between HR and LR in the control group, whereas nociception of LR tended toward an increase compared to HR in the FORM-exposed group. Results from the second experiment suggest that the effects of FORM exposure may be obscured by examining behavior in a heterogeneous population (HR and LR). This approach using animal models may help define neural substrates that mediate the amplification of responses of a subpopulation of individuals to chemicals in the environment. PMID- 8711730 TI - Adverse health effects of air pollutants in a nonsmoking population. AB - Utah Valley has provided an interesting and unique opportunity to evaluate the health effects of respirable particulate air pollution (PM10). Residents of this valley are predominantly nonsmoking members of the Church of Jesus Christ of Latter-day Saints (Mormons). The area has moderately high average PM10 levels with periods of highly elevated PM10 concentrations due to local emissions being trapped in a stagnant air mass near the valley floor during low-level temperature inversion episodes. Due to a labor dispute, there was intermittent operation of the single largest pollution source, an old integrated steel mill. Levels of other common pollutants including sulfur dioxide, ozone, and acidic aerosol are relatively low. Studies specific to Utah Valley have observed that elevated PM10 concentrations are associated with: (1) decreased lung function; (2) increased incidence of respiratory symptoms; (3) increased school absenteeism; (4) increased respiratory hospital admissions; and (5) increased mortality, especially respiratory and cardiovascular mortality. PMID- 8711731 TI - Kids are different: developmental variability in toxicology. AB - Data on the impact of many environmental compounds to human health is often lacking, particularly when considering the risk to the unborn and developing child. The stage of development of the individual at the time of exposure to a toxicant has not always been considered. For example, a higher percentage of ingested lead is absorbed from the gastrointestinal tract of infants than adults. Renal elimination also follows a developmental pattern, being very limited during the newborn period, increasing during infancy and childhood, and declining gradually after puberty. Genetic polymorphisms in metabolic enzyme activity add another dimension of variability. Depending on the particular chemical, this may serve as a protective factor or increase susceptibility to toxic effects (e.g. epoxide hydrolase and fetal hydantoin syndrome). Children also have distinctive behaviors and target organ susceptibilities that warrant special consideration. The consequences of developmental changes are well-known in medical practice, and many drug doses are modified based on age, liver, and renal function, and other factors that may influence pharmacokinetic behavior of drugs. There is a sizable body of such information available, in part, in the pediatric and clinical pharmacology and toxicology literature. The concept of the significance of developmental stage is becoming increasingly important in toxicological risk assessment as well. PMID- 8711732 TI - Comparison of four human studies of perinatal exposure to methylmercury for use in risk assessment. AB - Newer data from human epidemiologic studies of methylmercury (MeHg) poisoning in which perinatal exposure occurred are available from four distinct populations. The results of an Iraqi grain-consuming population are compared to results from studies performed in fish-consuming groups in the Faroe Islands, the Seychelles Islands and in Peruvian fishing villages. A comparison of the results indicate that the Iraqi population does not represent a sensitive subpopulation within a perinatal group, but rather the relative lower threshold identified from this study was the results of confounders. Use of this benchmark dose for regulatory purposes may place a severe limitation upon fish consumption in the United States that is not fully supported by the scientific data. PMID- 8711733 TI - Role of nutrition in the survival after hepatotoxic injury. AB - Nutritional status is an important factor in determining susceptibility to toxic chemicals. While macro and micronutrients may affect many aspects of Stage I and Stage II of toxicity, in this paper, the influence of macronutrients as sources of energy required for cell division and tissue repair mechanisms on the outcome of hepatic injury is discussed. Male Sprague-Dawley rats maintained on normal rodent chow and 15% glucose (as a source of energy for the centrilobular hepatocytes) in drinking water for 7 days experienced an increased lethality from structurally and mechanistically different centrilobular hepatotoxicants (acetaminophen, thioacetamide, chloroform and carbon tetrachloride), while male Sprague-Dawley (S-D) rats fed rat chow containing palmitic acid (PA, 8% w/w, as a source of energy for the periportal hepatocytes) and L-carnitine (LC, 2 mg/ml, as a mitochondrial carrier for the supplemented fatty acids) in drinking water for 7 days were protected from a LD100 dose (600 mg/kg, i.p.) of thioacetamide (TA). Indices of cell division revealed that cell cycle progression in the liver played a very critical role in determining the final outcome of hepatotoxic injury. These results confirmed our hypothesis that cell division and tissue repair play a critical role in survival after life-threatening hepatotoxic injury. Any manipulation directed towards altering a prompt and exacting compensatory cell division and tissue repair responses after hepatotoxic injury would also alter the final outcome of the toxicity. These studies indicate that the source of cellular energy can decisively influence the compensatory response of the target tissue to alter the outcome of hepatotoxic injury. Since nutritional status is known to vary widely among human populations, these could contribute enormously to susceptibility of human populations to toxic chemicals. PMID- 8711734 TI - Overview of diisocyanate occupational asthma. AB - Surveillance programs established around the world have determined that diisocyanate chemicals are the most common cause of occupational asthma. In the United States approximately 100,000 workers are exposed to these chemical compounds in the workplace each year and 5-10% of these workers will develop occupational asthma. There are no known reliable risk factors or biomarkers which can be used to predict which exposed worker will develop diisocyanate occupational asthma. Diisocyanate-occupational asthma workers manifest characteristic physiologic responses after specific bronchoprovocation which correlate with pathologic changes in their airways. However, the mechanism(s) for diisocyanate-occupational asthma remains unclear. Specific IgE antibody production to diisocyanates is found in only 10-30% of these workers. Bronchial biopsies and bronchoalveolar lavage have confirmed the presence of T-lymphocytes and eosinophils in the airways of these workers suggesting that T-cell mediated immune responses are more likely to play a central role in this disease. It is essential to diagnose diisocyanate-occupational asthma as early as possible in order to prevent or reduce the significant asthma morbidity associated with continuous long term exposure to these chemicals. Treatment of choice is removal of the worker from further exposure. Prospective studies evaluating larger populations of diisocyanate-exposed workers is essential for a better understanding of the pathogenesis and natural course of diisocyanate-occupational asthma. PMID- 8711735 TI - Characterization of histamine releasing factors in diisocyanate-induced occupational asthma. AB - Immunologic mechanisms contributing to diisocyanate-induced occupational asthma (OA) are poorly defined. There is a relatively low incidence of diisocyanate specific IgE antibody responses. The frequent occurrence of delayed onset asthmatic responses in workers with diisocyanate asthma suggests a role for cellular immune mechanisms. We have shown in vitro production of antigen-specific mononuclear cell-derived histamine releasing factors (HRF) by peripheral blood mononuclear cells (PBMCs) of workers with OA. Monocyte chemoattractant protein-1 (MCP-1) and RANTES (acronym for "regulated on activation normal T expressed and secreted") are chemokines found in PBMC supernatants that express HRF activity. Diisocyanate-exposed workers were tested for diisocyanate antigen-stimulated enhancement of HRF, MCP-1, and RANTES production in supernatants of PBMCs and for serum specific IgE and IgG antibody levels to diisocyanate antigens bound to human serum albumin (HSA). PBMCs of workers with diisocyanate OA showed significantly increased production of antigen-specific HRF activity and MCP-1 ( > 300 ng/ml) compared to diisocyanate-exposed asymptomatic workers (P < 0.05). Antigen-stimulated enhancement of MCP-1 mRNA was demonstrated by reverse transcription PCR. RANTES mRNA and chemokine secretion ( < 1 ng/ml) was also demonstrated in PBMCs, but did not show antigen enhancement in OA workers. Hapten specificity for the diisocyanate chemical to which a patient had been exposed was demonstrated for HRF enhancement and for IgG antibody reactions, but not for IgE reactions. HRF production was demonstrated in PBMC subpopulations, including lymphocytes and purified T cells. OA subjects showed increased CD8+ cells by immunofluorescence (mean CD4+: CD8+ = 1.2 +/- 0.2). The results suggest that diisocyanate antigen enhancement of HRF and MCP-1 production are stimulated by hapten-specific T cell reactions. Since a weak association has been found between IgE antibody synthesis and induction of diisocyanate OA, the role of T cell cytokines and chemokines in the pathogenesis of OA requires further investigation. PMID- 8711736 TI - Development and application of non-invasive biomarkers for carcinogen-DNA adduct analysis in occupationally exposed populations. AB - Biological monitoring of exposures to carcinogenic compounds in the workplace can be a valuable adjunct to environmental sampling and occupational medicine. Carcinogen-DNA adduct analysis has promise as a biomarker of effective dose if target organ samples can be obtained non-invasively. We have developed non invasive techniques using exfoliated urothelial and bronchial cells collected in urine and sputum, respectively. First morning urine samples were collected from 33 workers exposed to benzidine or benzidine-based dyes and controls matched for age, education, and smoking status. Sufficient DNA for 32P-postlabelling analysis was obtained from every sample. Mean levels of a specific DNA adduct (which co chromatographed with standard characterized by MS) were elevated significantly in the benzidine-exposed workers relative to controls. In addition, workers exposed to benzidine had higher adduct levels than those exposed to benzidine-based dyes. This study demonstrates the usefulness of these non-invasive techniques for exposure/effect assessment. To be useful in occupational studies, biomarkers must also be sensitive to exposure interventions. We have conducted topical application studies of used gasoline engine oils in mice and found that the levels of carcinogen-DNA adducts in skin and lung can be significantly lowered if skin cleaning is conducted in a timely manner. The combination of useful, non invasive techniques to monitor exposure and effect and industrial hygiene interventions can be used to detect and prevent exposures to a wide range of carcinogens including those found in used gasoline engine oils and jet exhausts. PMID- 8711737 TI - Quantitative cancer assessment for vinyl chloride: indications of early-life sensitivity. AB - Complementary sources of information are analyzed to characterize the early-life cancer risk from inhaling vinyl chloride. A study of partial-lifetime exposures suggests that the lifetime cancer risk depends on age at exposure, with higher lifetime risks attributable to exposures at younger ages. Studies of newborn animal exposures further demonstrate that a brief exposure in newborns can, by the end of life, induce a higher incidence of tumors compared to long-term exposure occurring later in life, including tumor types not induced by exposure later in life. An empirical, quantitative approach is used to model early-life sensitivity to inhaled vinyl chloride, supplementing conventional approaches for estimating the increased cancer risk from lifetime exposure. A single estimate is not presumed to apply to the entire population; instead, the new approach makes distinctions about the cancer risks for different population segments. This assessment shows one way such information might be analyzed, presented, and used to assess actual exposure situations. PMID- 8711738 TI - Possible health risks from low level exposure to beryllium. AB - The first case of chronic beryllium disease (CBD) at the Rocky Flats Environmental Technology Site (Rocky Flats) was diagnosed in a machinist in 1984. Rocky Flats, located 16 miles northwest of Denver, Colorado, is part of the United States Department of Energy (DOE) nuclear weapons complex. Research and development operations using beryllium began at Rocky Flats in 1953, and beryllium production operations began in 1957. Exposures could have occurred during foundry operations, casting, shearing, rolling, cutting, welding, machining, sanding, polishing, assembly, and chemical analysis operations. The Beryllium Health Surveillance Program (BHSP) was established in June 1991 at Rocky Flats to provide health surveillance for beryllium exposed employees using the Lymphocyte Proliferation Test (LPT) to identify sensitized individuals. Of the 29 cases of CBD and 76 cases of beryllium sensitization identified since 1991, several cases appear to have had only minimal opportunistic exposures to beryllium, since they were employed in administrative functions rather than primary beryllium operations. In conjunction with other health surveillance programs, a questionnaire and interview are administered to obtain detailed work and health histories. These histories, along with other data, are utilized to estimate the extent of an individual's exposure. Additional surveillance is in progress to attempt to characterize the possible risks from intermittent or brief exposures to beryllium in the workplace. PMID- 8711739 TI - Sampling and analysis of airborne resin acids and solvent-soluble material derived from heated colophony (rosin) flux: a method to quantify exposure to sensitizing compounds liberated during electronics soldering. AB - Components of colophony (rosin) resin acids are sensitizers through dermal and pulmonary exposure to heated and unheated material. Significant work in the literature identifies specific resin acids and their oxidation products as sensitizers. Pulmonary exposure to colophony sensitizers has been estimated indirectly through formaldehyde exposure. To assess pulmonary sensitization from airborne resin acids, direct measurement is desired, as the degree to which aldehyde exposure correlates with that of resin acids during colophony heating is undefined. Any analytical method proposed should be applicable to a range of compounds and should also identify specific compounds present in a breathing zone sample. This work adapts OSHA Sampling and Analytical Method 58, which is designed to provide airborne concentration data for coal tar pitch volatile solids by air filtration through a glass fiber filter, solvent extraction of the filter, and gravimetric analysis of the non-volatile extract residue. In addition to data regarding total soluble material captured, a portion of the extract may be subjected to compound-specific analysis. Levels of soluble solids found during personal breathing zone sampling during electronics soldering in a Naval Aviation Depot ranged from below the "reliable quantitation limit" reported in the method to 7.98 mg/m3. Colophony-spiked filters analyzed in accordance with the method (modified) produced a limit of detection for total solvent-soluble colophony solids of 10 micrograms/filter. High performance liquid chromatography was used to identify abietic acid present in a breathing zone sample. PMID- 8711740 TI - Exposure-response functions in Air Force toxic risk modeling. AB - A new methodology for estimating the probabilistic risk from acute toxic exposures is planned as a support tool for the Air Force at the Eastern and Western Ranges. Two such methodologies are programs entitled the Launch Area Toxic Analysis program (LATRA) and the Cold Spill Toxic Risk Analysis program (COSTRA). These programs combine probabilistic models of an accident (when applicable), release cloud formation and dispersion (appropriate to the toxic substance and accounting for meteorological conditions), and new exposure response functions (ERFs) for sensitive and normal exposed populations. These ERFs, anchored on specific exposure standards, estimate the probability of a given severity of health effect in a particular population as a function of the concentration or dose to which it is exposed. The further development and acceptance of these ERFs by the toxicology community, especially for different sensitivities, are key concerns addressed in this paper. PMID- 8711741 TI - Obtaining information about susceptibility from the epidemiological literature. AB - Whether people become ill after encountering environmental pollutants depends on the magnitude of their exposure and their capacity to respond. Exposure and intrinsic response capabilities vary within the population. Those that become ill when the general population remains largely unaffected are considered to be highly susceptible. The U.S. Environmental Protection Agency (USEPA), responsible for protecting the public from environmental pollutants, has developed risk assessment procedures to assist in evaluating the likelihood of health effects. However, the Agency's ability to evaluate the risk faced by highly susceptible populations is often hindered by the paucity of adequate health effects data. Response variability can be assessed with animal models and human epidemiological studies. Although animal models are useful when evaluating the effect of gender and developmental stage on susceptibility, inbred rodent strains underestimate the genetic and lifestyle-induced variability in susceptibility found in human populations. Epidemiological approaches are the preferred source of information on variability. This paper reviews the epidemiological literature from the perspective of a risk assessor seeking data suitable for estimating the risk to highly susceptible populations. Epidemiological approaches do not measure the full range of population response variability. Rather, "susceptibility factors" are evaluated either as risk factors or by focusing on the susceptible population, e.g. children. Susceptibility factors due to genetics, developmental stage, gender, ethnicity, disease state and lifestyle are most frequently encountered. Often, the information describing the health impact of the susceptibility factor is incomplete due to, (1) a failure to consider factors modifying susceptibility; (2) inadequate exposure data; (3) a failure to evaluate the health impact of the susceptibility factor. In addition, for a given exposure agent, several susceptibility factors may be relevant. While incomplete data describing susceptibility factors limits the opportunity for quantitative estimations of risk, available information can supplement qualitative evaluations and risk management. PMID- 8711742 TI - Evaluating human variability in chemical risk assessment: hazard identification and dose-response assessment for noncancer oral toxicity of trichloroethylene. AB - Human variability can be addressed during each stage in the risk assessment of chemicals causing noncancer toxicities. Noncancer toxicities arising from oral exposure to trichloroethylene (TCE) are used in this paper as a case study for exploring strategies for identifying and incorporating information about human variability in the chemical specific hazard identification and dose-response assessment steps. Toxicity testing in laboratory rodents is the most commonly used method for hazard identification. By using animal models for sensitive populations, such as developing fetuses, testing can identify some potentially sensitive populations. A large variety of reproductive and developmental studies with TCE were reviewed. The results were mostly negative and the limited positive findings generally occurred at doses similar to those causing liver and kidney toxicity. Physiologically based pharmacokinetic modeling using Monte Carlo simulation is one method for evaluating human variability in the dose-response assessment. Three strategies for obtaining data describing this variability for TCE are discussed: (1) using in vivo human pharmacokinetic data for TCE and its metabolites, (2) studying metabolism in vitro, and (3) identifying the responsible enzymes and their variability. A review of important steps in the metabolic pathways for TCE describes known metabolic variabilities including genetic polymorphisms, enzyme induction, and disease states. A significant problem for incorporating data on pharmacokinetic variability is a lack of information on how it relates to alterations in toxicity. Response modeling is still largely limited to empirical methods due to the lack of knowledge about toxicodynamic processes. Empirical methods, such as reduction of the No-Observed Adverse-Effect-Level or a Benchmark Dose by uncertainty factors, incorporate human variability only qualitatively by use of an uncertainty factor. As improved data and methods for biologically based dose-response assessment become available, use of quantitative information about variability will increase in the risk assessment of chemicals. PMID- 8711743 TI - Investigation of the impact of pharmacokinetic variability and uncertainty on risks predicted with a pharmacokinetic model for chloroform. AB - A sensitivity and uncertainty analysis was performed on the Reitz et al. (Toxicol. Appl. Pharmacol., 1990: 105, 443) physiologically based pharmacokinetic (PBPK) risk assessment model for chloroform. The analytical approach attempted to separately consider the impacts of interindividual variability and parameter uncertainty on the predicted values of the dose metrics in the model, as well as on liver cancer risk estimates obtained with the model. An important feature of the analytical approach was that an attempt was made to incorporate information on correlation between important parameters, for example, the observed correlation between total blood flow and alveolar ventilation rate. Using the published PBPK model for chloroform, the best estimate of the average population risk based on the preferred pharmacodynamic dose metric (PTDEAD), representing cell death, is 9.2 x 10(-7); this estimate is more than 500-fold lower than the risk estimate of 5.3 x 10(-4) based on an alternative pharmacokinetic dose metric (AVEMMB), which represents tissue adduct formation. However, when interindividual variability was considered the range of individual risks (from the 5th to the 95th percentile of the population) predicted with PTDEAD was extremely broad (from 3.0 x 10(-13) to 3.2 x 10(-4)), while individual risks predicted with AVEMMB only varied over a factor of four (from 1.9 x 10(-4) to 7.4 x 10(-4)). As a result, the upper 95th percentile of the distribution of individual risk estimates based on the preferred cell death metric were within a factor of three of the 95th percentile for the pharmacokinetic alternative. The crucial factor with respect to the much greater variability of chloroform risk estimates based on cell death is that the dose metric, PTDEAD, is exquisitely sensitive to variation of the parameters in the model defining the response of cells to the cytotoxicity of chloroform. Unfortunately, these key parameters are also highly uncertain, as well as strongly correlated. As a result it proved impossible to accurately quantify the additional impact of parameter uncertainty on the dose metrics and risk estimates for chloroform. In general, however, the approach used in this study should be useful for differentiating the impact of interindividual variability and parameter uncertainty on PBPK-based risk assessments of other chemicals where the sensitivity, uncertainty, and correlation of the key parameters are more limited. PMID- 8711744 TI - Efficient tissue repair underlies the resiliency of postnatally developing rats to chlordecone + CCl4 hepatotoxicity. AB - It is often assumed that at a younger age populations are at higher risk of toxic effects from exposure to toxic chemicals. Recent studies have demonstrated that neonate and postnatally developing rats are resilient to a wide variety of structurally and mechanistically dissimilar hepatotoxicants such as galactosamine, acetaminophen, allyl alcohol, and CCl4. Most interestingly, young rats survive exposure to the lethal combination of chlordecone (CD) + CCl4 known to cause 100% mortality in adult male and female rats. In a study where postnatally developing (20- and 45-day), and adult (60-day) male Sprague Dawley rats were used, administration of CCl4 (100 microliters/kg, i.p.) alone resulted in transient liver injury regardless of age as indicated by plasma alanine transaminase (ALT), sorbitol dehydrogenase (SDH) levels and histopathological lesions. In CD-pretreated rats, CCl4-induced toxicity progressed with time culminating in 25 and 100% mortality by 72 h after CCl4 in 45- and 60-day rats, respectively, in contrast to regression of CCl4-induced toxicity and 0% mortality in 20-day rats. [3H]Thymidine (3H-T) incorporation and proliferating cell nuclear antigen (PCNA) studies revealed an association between delayed and diminished DNA synthesis, unrestrained progression of liver injury, and animal death. Time course studies revealed that the loss of resiliency in the two higher age groups might be due to inability to repair the injured liver rather than due to infliction of higher injury. Intervention of cell division in 45-day CD rats by colchicine (CLC, 1 mg/kg, i.p.) 30 h after CCl4 challenge increased mortality from 25 to 85%, confirming the importance of stimulated tissue repair in animal survival. In contrast, efficient and substantial DNA synthesis observed in 20-day rats allows them to limit further progression of liver injury, thereby leading to full recovery of this age group with 0% mortality. Examination of growth factors and proto-oncogene expression revealed a 3- and 3.5-fold increase in transforming growth factor-alpha (TGF-alpha) and H-ras mRNA expressions, respectively, coinciding with maximal hepatocyte DNA synthesis in 20-day normal diet (ND) rats, as opposed to only 2- and 2.5-fold increases observed in 60-day ND rats, respectively. Increased expression of c-fos (10-fold) in 20-day rats occurred 1 h after CCl4 compared to less than a 2-fold increase in 60-day rats. These findings suggest that prompt stimulation of tissue repair permits efficient recovery from injury during early postnatal development of rats. PMID- 8711745 TI - Risk assessment policy for evaluating reproductive system toxicants and the impact of responses on sensitive populations. AB - Risk assessment policy for evaluating environmental chemicals for their potential to produce reproductive system failures is similar to policy for evaluating cancer-causing effects. The objective of reproductive system risk assessment is to expand on the test standards that primarily focus on fertility endpoints and birth defects by using mechanism-of-action studies and quantitative risk assessment methods. An understanding of the sensitivity of reproductive system insult between animal species and from animal models to man is critical to developing risk assessment policy and test standards. The reproductive process is complex and involves a number of maturation and sex cell development processes. Sensitivity to insult varies throughout this process, especially during, (1) the development of the conceptus, sperm and ova, (2) fertilization, (3) implantation, and (4) puberty. Reproductive failure has many causes and clinical effects. Risk assessment policy is directed toward reducing the uncertainty associated with the cause by providing a guide to understanding how dose, duration, and characteristics of the reproductive toxicant affect the reproductive process. PMID- 8711746 TI - Use of physiologically based pharmacokinetic modeling to investigate individual versus population risk. AB - Because of the heterogeneity of the human population, it is generally expected that there will be a broad range of observed susceptibilities to the biological effects of exposure to chemicals or drugs. Often it is possible to distinguish specific classes of individuals, such as infants or the elderly, who appear to be more susceptible to a specific effect. Non-cancer risk assessment often address this variability by dividing the experimentally determined acceptable exposure level by an uncertainty factor of 10 to protect sensitive individuals; cancer risk assessments typically do not address this issue in any quantitative fashion. Physiologically based pharmacokinetic (PBPK) modeling provides the capability to quantitatively describe the potential impact of pharmacokinetic factors on the variability of individual risk. In particular, PBPK models can be used to determine the impact of differences in key metabolism enzymes, whether due to multiple genotypic expression, such as cytochrome P450 polymorphisms, or just due to normal variation in enzyme activities within the general population. Other potential modulators of sensitivity which can be addressed quantitatively with a PBPK model include physical condition, level of activity, disease states, age, hormonal status, and interactions with other chemicals and drugs. In each case, the PBPK model provides a quantitative structure for determining the effect of these various factors on the relationship between the external (environmental) exposure and the internal (biologically effective) target tissue exposure. When coupled with Monte Carlo analysis, the PBPK model provides a method to assess the quantitative impact of these sources of variability on individual risk (as opposed to average population risk) by comparing model-predicted risks over the distribution of individual parameter values. PMID- 8711747 TI - Interindividual variations in susceptibility and sensitivity: linking risk assessment and risk management. AB - In the past few years, our knowledge of mammalian genomes has increased enormously. Our understanding of the molecular basis of the normal cellular processes of DNA replication and repair and cell cycle control, together with how their fidelity malfunctions as part of tumor development, has increased in parallel. This has led to a clearer appreciation that there are subpopulations that have been generically described as being genetically or otherwise susceptible to the induction of cancer or birth defects. The term susceptibility is a default option, since there clearly will be a very broad range of sensitivities among the so-called susceptible populations, dependent upon the specific underlying mechanism. This could lead to the conduct of risk assessments for each specific situation, involving both genotypes of individuals and agents of concern. This would ideally take into account the effects on response of various modifying factors, genetic and other. One advantage to be gained from this approach is the ability to determine if a particular susceptibility places subpopulations at extreme risk as compared to the overall normal distribution of risk in the population, or whether such a susceptible population presents a slight extension of the upper bound of the risk distribution or lies within the normal distribution. In addition, the specific mechanism of the susceptibility as related to exposure scenarios and the magnitude and demographics of the susceptible populations need to be taken into account. Thus, the management of risk has to be linked to the specific risk assessment. For many of the so-called susceptible populations an uncertainty factor of less than 10, even including 1, would be predicted to bring the risk within the normal distribution. It is hoped that as more mechanistic information on susceptibility becomes available and a specific risk can be defined, the practice of risk management will be considerably improved. PMID- 8711748 TI - Phenotypic variation in xenobiotic metabolism and adverse environmental response: focus on sulfur-dependent detoxification pathways. AB - Proper bodily response to environmental toxicants presumably requires proper function of the xenobiotic (foreign chemical) detoxification pathways. Links between phenotypic variations in xenobiotic metabolism and adverse environmental response have long been sought. Metabolism of the drug S-carboxymethyl-L-cysteine (SCMC) is polymorphous in the population, having a bimodal distribution of metabolites, 2.5% of the general population are thought to be nonmetabolizers. The researchers developing this data feel this implies a polymorphism in sulfoxidation of the amino acid cysteine to sulfate. While this interpretation is somewhat controversial, these metabolic differences reflected may have significant effects. Additionally, a significant number of individuals with environmental intolerance or chronic disease have impaired sulfation of phenolic xenobiotics. This impairment is demonstrated with the probe drug acetaminophen and is presumably due to starvation of the sulfotransferases for sulfate substrate. Reduced metabolism of SCMC has been found with increased frequency in individuals with several degenerative neurological and immunological conditions and drug intolerances, including Alzheimer's disease, Parkinson's disease, motor neuron disease, rheumatoid arthritis, and delayed food sensitivity. Impaired sulfation has been found in many of these conditions, and preliminary data suggests that it may be important in multiple chemical sensitivities and diet responsive autism. In addition, impaired sulfation may be relevant to intolerance of phenol, tyramine, and phenylic food constituents, and it may be a factor in the success of the Feingold diet. These studies indicate the need for the development of genetic and functional tests of xenobiotic metabolism as tools for further research in epidemiology and risk assessment. PMID- 8711749 TI - Human interindividual variability in susceptibility to toxic effects: from annoying detail to a central determinant of risk. AB - It is unusual to find variability issues as the central focus of a scientific conference. The discussion below first suggests why variability has often been an "annoying detail" in both basic animal toxicology and the human testing of new drugs. Then it gives some reasons why improved quantitative variability information is likely to be important. Better definition of the sources and magnitude of variability in susceptibility in the human population is a central issue for, (1) making more quantitative estimates of both cancer and non-cancer risks from occupational and environmental exposures, and (2) designing protocols for the use of drugs that maximize benefits for the risks incurred in a diverse patient population. Finally, it offers some suggestions about how better variability information is to be obtained and/or extracted from existing information. PMID- 8711750 TI - Chemical sensitivity: symptom, syndrome or mechanism for disease? AB - Several different meanings have been attached to the term "chemical sensitivity" by those who use it. Feeling ill from odors is a symptom reported by approximately one-third of the population. The syndrome of chemical sensitivity, frequently called "Multiple Chemical Sensitivity" or "MCS" has been the subject of three federally-sponsored workshops; at least five different case definitions for research on MCS have been proposed. In contrast, the hypothesis that chemical sensitivity may be a mechanism for disease posits that a broad spectrum of "recognized" chronic illnesses, ranging from asthma and migraine to depression and chronic fatigue, may be the consequence of environmental chemical exposures. According to this theory, a two-step process occurs: (1) an initial salient exposure event(s) (for example, a one-time, intermittent, or continuous exposure to pesticides, solvents, or air contaminants in a sick building) interacts with a susceptible individual, causing loss of tolerance for everyday, low level chemical inhalants (car exhaust, fragrances, cleaning agents), as well as for foods, drugs, alcohol, and caffeine; (2) thereafter, such common, formerly well tolerated substances trigger symptoms, thus perpetuating illness. "Masking" (acclimatization, apposition, and addiction) may hide these exposure-symptom relationships, thus obfuscating the environmental etiology of the illness. Accumulating clinical observations lend credence to a view of chemical sensitivity as an emerging theory of disease causation and underscore the need for its testing in a rational, scientific manner. While chemical sensitivity may be the consequence of chemical exposure, the term "toxicant-induced loss of tolerance" more fully describes the two-step process under scrutiny. PMID- 8711751 TI - Sensitization induced by kindling and kindling-related phenomena as a model for multiple chemical sensitivity. AB - It has been suggested that the neurobehavioral dysfunction observed in persons presenting with symptoms of Multiple Chemical Sensitivity (MCS) syndrome involves sensitization of neural circuits. Two hypotheses for the route of exposure in induction of neural sensitization in MCS are: (a) direct chemical stimulation of olfactory processes, or (b) general systemic response to inhaled chemicals. In either case, the mechanism of action may involve chemical kindling or kindling related phenomena. A neural sensitization mechanism based on kindling or kindling related phenomena is attractive and has been previously demonstrated in both in vitro and in vivo animal models. Without a testable animal model for chemically mediated induction of MCS, however, any argument that MCS is mediated by kindling or kindling-related phenomena is reduced to the circular argument "the mechanism of sensitization is sensitization." The present survey provides an overview of the experimental paradigms that result in sensitization, differentiated on the basis of probable neurophysiological and neurochemical mechanisms. Neurophysiological potentiation, electrical kindling, chemical kindling and behavioral sensitization are evaluated and discussed in relationship to MCS. PMID- 8711752 TI - Sir Joseph Fayrer MD FRS (1824-1907) Indian Medical Service: snakebite and mortality in British India. AB - Whilst Professor of Surgery at the Calcutta Medical College, Dr Fayrer studied all aspects of snake poisoning. His work, The Thanatophidia of India being a Description of the Venomous Snakes of the Indian Peninsula, with an Account of the Influence of their Poison of Life; and a Series of Experiments was published in 1872. Collating information from official Government records of death due to snakebite, Fayrer recorded a mortality of 11,416 persons for the year 1869 in the Presidency of Bengal and estimated a mortality of more than 20,000 for all British India. In 1872, working in London with Lauder Brunton, Fayrer studied the physiological action on Naja naja venom in more detail. Respiratory paralysis was due to a curare-like blockade of the neuromuscular junction, and they believed, a central effect also; artificial respiration was ineffective in producing recovery. A large dose of cobra venom was observed to induce tetanic contraction of the heart both in vivo and in vitro. As President of the Medical Board of the India Office in London from 1873 to 1895, Sir Joseph Fayrer was much concerned with cholera epidemics which resulted in an average mortality of over 200,000 persons annually. In 1885, he represented the Indian Government at the fourth International Conference on Cholera, a fore-runner of the World Health Organization. PMID- 8711753 TI - Synaptosomal binding of 125I-labelled daboiatoxin, a new PLA2 neurotoxin from the venom of Daboia russelli siamensis. AB - Daboiatoxin (DbTx), the PLA2 neurotoxin from Daboia russelli siamensis venom, was shown to bind specifically and saturably to rat cerebrocortical synaptosomes and synaptic membrane fragments. Two families of binding sites were detected by equilibrium binding analysis in the presence and absence of Ca2+. Scatchard analysis of biphasic plateaus revealed Kdl 5 nM and Bmax1, 6 pmoles/mg protein, and Kd2 80 nM and Bmax2 20 pmoles/mg protein, respectively, for the high- and low affinity binding sites. The binding of 125I-DbTx to synaptosomes did not show marked dependence on Ca2+, Mg2+, Co2+ and Sr2+. Native DbTx was the only strong competitor to 125I-DbTx synaptosomal binding (IC50 12.5 nM, KI 5.5 nM). Two other crotalid PLA2 neurotoxins, crotoxin CB and mojave toxin basic subunit, and nontoxic C. Atrox PLA2 enzyme, were relatively weaker inhibitors, while two viperid PLA2 neurotoxins, ammodytoxin A and VRV PL V, were very weak inhibitors. Crotoxin CA was a poor inhibitor even at microM concentrations, whereas no inhibitory effect at all was observed with crotoxin CACB, ammodytoxin C, VRV PL VIIIa, taipoxin, beta-bungarotoxin, or with PLA2 enzymes from N. naja venom, E. schistosa venom, bee venom and porcine pancreas. All other pharmacologically active ligands examined (epinephrine, norepinephrine, histamine, choline, dopamine, serotonin, GABA, naloxone, WB-4101, atropine, hexamethonium and alpha bun-garotoxin) also failed to interfere with 125I-DbTx binding. As those competitors that showed partial inhibition were effective only at microM concentration range compared to the Kd (5 nM) of 125I-DbTx synaptosomal binding, DbTx could well recognize a different neuronal binding site. Rabbit anti-DbTx polyclonal antisera completely blocked the specific binding. When a range of Ca2+ and K+ channels modulators were examined, Ca2+ channel blockers (omega-conotoxins GVIA and MVIIC, taicatoxin, calciseptine and nitrendiprene) did not affect the binding even at high concentrations, while charybdotoxin was the only K+ channel effector that could partially displace 125I-DbTx synaptosomal binding amongst the K+ channel blockers tested (apamin, dendrotoxin-I, iberiotoxin, MCD-peptide, 4 aminopyridine and tetraethylammonium), suggesting that neither K+ nor Ca2+ channels are associated with DbTx binding sites. PMID- 8711754 TI - Pathological changes induced by an acidic phospholipase A2 from Ophiophagus hannah venom on heart and skeletal muscle of mice after systemic injection. AB - An acidic phospholipase A2 (OHV A-PLA2) isolated from the venom of the king cobra (Ophiophagus hannah) was tested for its ability to cause pathological changes to myocardium, skeletal muscle and cardiac ganglia. White mice were injected intravenously with dose of 8 mg/kg or 4 mg/kg of OHV A-PLA2 and tissue samples were taken at 6 or 24 hr. Light microscopic examination failed to show significant changes in cardiac muscle and ganglia. Skeletal muscle showed myofibre degeneration and necrosis. Electron microscopic study revealed myodegeneration in cardiac and skeletal muscles, and reduction in synaptic vesicle population of preganglionic nerve terminals in cardiac ganglia. Ultrastructural changes in tissues were dose related. The lower dose (4 mg/kg) of OHV A-PLA2 produced mild myocardial changes, the myofilaments were intact but contracted, and the A band and I band were skewed. OHV A-PLA2 caused myocardial degeneration at a higher dose of 8 mg/kg. The changes included dissolution of actin and myosin filaments, dilatation and disorganization of sarcoplasmic reticulum and degeneration of mitochondria. The skeletal muscle lesions were more severe than the myocardial changes. Some of the myofibrils were severely disorganized and lack typical striated appearance, sarcomeres disrupted, most of mitochondria were vesiculated and destroyed. PMID- 8711755 TI - Facile production of native-like kappa-bungarotoxin in yeast: an enhanced system for the production of a neuronal nicotinic acetylcholine receptor probe. AB - Research on the mammalian central nervous system had been hindered by the limited number and meager supply of naturally occurring toxins that can be used as pharmacological reagents. The kappa-neurotoxins in particular are not found abundantly in nature and are difficult to obtain and isolate in quantities sufficient for research purposes. Here we report the expression and isolation of relatively large quantities of the kappa-neurotoxin, kappa-bungarotoxin, in an active form using a yeast, Pichia pastoris, expression system. The resultant product of the expression system has a short amino-terminal amino acid extension relative to venom-derived kappa-bungarotoxin, but is equivalent to the native toxin in physical and biological properties, as judged by the CD spectra, the ability to form dimers in solution, and the activity on chick ciliary ganglia. The yeast system produces approximately 0.2 mg from a 2 liter culture and the purification takes approximately 2 days. In contrast, E. coli, the only other available expression system for this toxin, produces one-fifth to one-half as much active material from a 5 liter high-density fermentation and the resulting protein takes over a week to purify. No high mol. wt disulfide-bonded aggregates were found in the yeast expression system product, indicating that the product is that of a biologically assisted folding process. This has significant implications not only for the efficient production of native toxin but also for the production of mutant proteins to study the structure-function relationship in these proteins. PMID- 8711756 TI - High incidence of bites and stings by snakes and other animals among rubber tappers and Amazonian Indians of the Jurua Valley, Acre State, Brazil. AB - Among forest-dwelling Amazonian Indians and rubber tappers (seringueiros) of the Jurua valley in Acre State, north-western Brazil, snakebite is an important cause of morbidity and death. Overall, 13% of a surveyed population had been bitten during their lifetime. Seventeen per cent of Katukina Indians, but only 8% of Ashaninkas, had been bitten by snakes reflecting, perhaps, different levels of traditional knowledge of the forest and its dangers. Most bites occurred in the jungle or on jungle trails (56%), while people were working (41%) or walking (26%), and were inflicted on the feet (54%). Ninety per cent of bite victims received treatment, usually traditional (93%); the majority (80%) recovered fully. Mortality was estimated at about 400 deaths per 100,000 population per lifetime. Bites and stings from other venomous forest and river animals, especially the freshwater sting ray (Potamotrygon sp.), were also extremely common. One death from an ant bite was recorded. PMID- 8711757 TI - Effect of 3,4-diaminopyridine on rat extensor digitorum longus muscle paralyzed by local injection of botulinum neurotoxin. AB - The actions of the K+ channel blocker, 3,4-diaminopyridine (3,4-DAP), were studied in the rat extensor digitorum longus (EDL) muscle following local inhibition of neuromuscular transmission by botulinum neurotoxin (BoNT). Local paralysis of the EDL muscle was induced by s.c. injections of BoNT serotypes A, B, E or F over the anterior tibialis muscle. One to 14 days later, the rats were anesthetized with urethane, and isometric twitch tensions following stimulation of the peroneal nerve were measured in situ. Muscles were paralyzed within 24 hr of administration of 5 mouse LD50 units (U) of BoNT/A and remained inhibited for the entire 14-day period of observation. Similar levels of inhibition, but of shorter duration, were observed after local injection of 20 U of BoNT/E, 10(4) U of BoNT/B or 20 U of BoNT/F. 3,4-DAP (4 mg/kg, i.v.) potentiated twitch tensions markedly in BoNT/A intoxicated muscle. The increase in tension developed rapidly (halftime = 5.81 +/- 0.6 min), persisted for approximately 1 hr, then decayed slowly with a halftime of 25.2 +/- 4.6 min. Subsequent administration of 3,4-DAP restored tensions to the original maxima, and this procedure could be repeated up to eight times with no decrement. The action of 3,4-DAP was comparable when given 1, 2, 3 or 7 days after BoNT/A and enhanced when administered 14 days after toxin injection. 3,4-DAP was less effective in reversing BoNT/E-induced muscle paralysis and nearly ineffective in antagonizing the paralytic actions of BoNT/B or BoNT/F. The results indicate that 3,4-DAP is of benefit in BoNT/A and BoNT/E intoxication, but is of marginal value after exposure to serotypes B and F. PMID- 8711758 TI - Cloning and characterization of the genomic region encoding toxin IV-5 from the scorpion Tityus serrulatus Lutz and Mello. AB - By means of PCR and using synthetic oligonucleotides designed from the reported cDNA, we amplified the gene that codes for toxin IV-5 from the Brazilian scorpion Tityus serrulatus. The analysis of the nucleotide sequence shows that the amplified genomic region is composed of 659 base pairs (bp) comprising two exons (28 and 284 bp) and an intron of 347 bp interrupting the region that encodes the signal peptide of the precursor toxin. Based on these findings a model for the structural organization of scorpion toxin genes is proposed. PMID- 8711759 TI - New procedures and parameters for better evaluation of Androctonus australis garzonii (Aag) and Buthus occitanus tunetanus (Bot) scorpion envenomations and specific serotherapy treatment. AB - New procedures describing intoxication with variable amounts of scorpion venoms (from 1 to 5 LD50) allowed us to introduce new parameters to evaluate Aag and Bot envenomations. Significant differences between the fatal limit time (FLT) and the last mortality time (LMT) were observed when the amount of Aag and Bot venom injected was equal to 1 LD50 and equal to or higher than 2 LD50. For Aag and Bot, the percentage of the fast mortality (FM) and the delayed mortality (DM) varied conversely when the amount of injected venom increased from 1 to 5 LD50. The relationship between the venom LD50 (from 2 to 20), the median protective dose (PD50) and the neutralizing activity of specific antivenom have been established. PD50 increased in a parallel manner with LD50. The neutralizing titres (LD50/ml) of Aag antivenom decreased from 74 +/- 3 to 44 +/- 2 and that of Bot antivenom from 52 +/- 2 to 36 +/- 1 when the number of LD50 injected increased from 2 to 20. Antivenom potency was evaluated using different protocols based on the presence or the absence of preincubation of the venom with the antivenom. In experiments where venom and antivenom were simultaneously but immediately injected, PD50 were twice as high as those found when venom and antivenom were preincubated (30 min at 37 degrees C). On the contrary, the corresponding neutralizing titres were two times lower. In an attempt to simulate accidental envenomations and subsequent serotherapy, Aag and Bot venom (4 LD50) were subcutaneously injected and the appropriate PD50S of antivenom were intravenously administered at different time intervals after envenomation. When the time of antivenom administration was shorter than the FLT, all envenomed mice might be protected by increasing volume of antivenom. However, when the antivenom is injected closer to the FLT only 50 to 60% of mice envenomed, respectively, by Aag and Bot could be saved even when more than 5 PD50 were injected. PMID- 8711760 TI - Immunochemical characterization of type A botulinum neurotoxin in its purified and complexed forms. AB - Immunochemical reactivities of type A botulinum neurotoxin to polyclonal antibodies raised against the neurotoxin complex toxoid have been investigated using enzyme-linked immunosorbent assay (ELISA) and with a fiberoptic immunosensor. The complex contains a group of complexing proteins in addition to the neurotoxin itself. Neurotoxin specific immunoglobulin G (IgG), purified from the IgG raised against the complex using an affinity column, showed a two-fold increase in reactivity with purified neurotoxin compared to the neurotoxin in the complex. Antibodies against the whole complex reacted approximately five times better with the complex than with the neurotoxin, suggesting that the detection of the neurotoxin complex may be more sensitive. Considering the fact that the amount of the complexing proteins and neurotoxin appears to be in a 4:1 ratio, a five-fold higher reactivity could suggest a 25-fold higher detectability of the neurotoxin in the complex. ELISA binding curves of complexing proteins and purified neurotoxin with antibodies raised against the whole complex indicated the complexing proteins to have significantly higher antigenicity. Furthermore, IgG fraction with or without the neurotoxin specific antibodies reacted almost equally to the neurotoxin complex, again suggesting higher immunogenicity of the complexing proteins. Increased binding of the complexing proteins versus the purified neurotoxin to antibodies against the complex and thus immunogenicity was also observed in the binding curves generated using a fiberoptic immunosensor. PMID- 8711762 TI - Bibliography of toxinology. PMID- 8711761 TI - The coronary vasoconstrictor action of extract IV from the dahlia sea anemone Tealia felina L. AB - Extract IV, a partially purified preparation of the toxin from the sea anemone Tealia felina, produced marked bradycardia and arrhythmias in the rat in vivo. In the Langendorff rat heart preparation perfused at constant pressure extract IV (0.0224 AU/ml) reduced the force of contraction by 81.3 +/- 7.2%, n = 6, and the coronary flow by 82.0 +/- 4.7%, n = 5. When the preparation was perfused at constant flow rate, extract IV (0.072 AU/ml) increased the coronary circulation resistance from 3.76 +/- 1.09 (control) to 36.94 +/- 12.26 mmHg/ml/min. The force of contraction was also markedly reduced. The extract produced bradycardia in preparations perfused at constant pressure but not in those perfused at constant flow rate. The extract did not affect isolated atria preparations. It was concluded that the bradycardia produced in vivo and in preparations perfused in vitro at constant pressure was probably secondary to the coronary vasospasm produced, which could contribute to the cardiotoxicity of the extract. PMID- 8711763 TI - Respiratory hypersensitivity and environmental factors: East and West Germany. AB - We assessed the prevalence of asthma and allergic disorders in 9-11 year old children in Leipzig and Halle in East Germany, and Munich, West Germany. Both East German cities were heavily polluted due to private coal burning and industrial emissions, whereas Munich has moderate industry but heavy car traffic. All fourth grade pupils in Munich (n = 7445) were compared with those in Leipzig, 1990 (n = 1429) and Leipzig and Halle, 1991 (n = 3105). The prevalence of hay fever and skin test reactivity to common aeroallergens was considerably higher in West Germany as compared to East Germany. Furthermore, the prevalence of asthma was also higher in the West German study area. However, when atopy was taken into account, there was no longer a significant difference in the prevalence of asthma between the two parts of the country. PMID- 8711764 TI - Environmental factors and respiratory hypersensitivity: the Americas. AB - Diverse environmental exposure profiles exist in the Americas because of widely different climates, ambient pollutants, and bioaerosols in these continents. This paper reviews selected studies from the Americas that support the broad hypothesis that environmental factors contribute to respiratory hypersensitivity. Processes influenced by environmental factors include primary immunologic sensitization, the development and exacerbation of specific immunologic diseases and the activation of nonspecific mechanisms with tissue inflammation, injury and remodeling. Endpoints resulting from these processes include respiratory symptoms, diseases such as asthma, with measures of disease severity including medication use and hospitalization rates, and death due to cardiorespiratory disease. Studies associate sensitization rates to specific allergens with environmental factors such as humidity and indices of allergen exposure. Regional variation occurs with exposure to outdoor source pollutants such as ozone, but varies by household to bioaerosols such as dust mite, cat or cockroach allergen. Indoor allergens are associated with asthma while outdoor allergens are associated with allergic rhinitis. In a national survey, the atopic sensitization rate in the USA increased with urban residence (defined as towns of population > 2500) and varied by region. Controlled human challenge studies show that ozone increases the response of allergic subjects to allergen. Increased ambient photochemical pollution concentrations, of which ozone is an important component, are associated with increased emergency room visits for asthma in cities such as Toronto, New York, Atlanta, and Mexico City. In Sao Paolo, Brazil, mortality due to childhood respiratory disease was influenced by the ambient levels of NO2. Epidemiologic studies including the recent meta-analysis of a large, longitudinal study population associate ambient concentrations of particulate matter < 10 microns and respiratory symptoms, disease severity and increased cardiorespiratory deaths. Toxicology studies show that individual variation in responsiveness is important in nonspecific inflammatory responses to irritant pollutants such as ozone and environmental tobacco smoke. These studies indicate that environmental factors influence primary allergen sensitization, secondary allergic responses, the activation of nonspecific inflammatory responses, and the severity of respiratory diseases, including asthma. PMID- 8711765 TI - Exposure to environmental chemicals relevant for respiratory hypersensitivity: global aspects. AB - Trends of air pollutant concentrations that influence the incidence of respiratory diseases and might be influential for respiratory hypersensitivity are presented and discussed. Data for these trends are collected in UNEP/WHO's Global Environmental Monitoring System GEMS/Air which recently has been revitalized and redirected to become a tool for providing the information necessary for rational air quality management. Data are interpreted with respect to WHO's most recent air quality guidelines, which are an update of the air quality guidelines for Europe published in 1987. A WHO study on the global disease burden and the contribution of air pollution to it is discussed and global estimates of mortality resulting from air pollution through sulphur dioxide and suspended particulate matter, by different economic regions, are given. Starting from the air quality guidelines still valid at the time of publication of the update, about 460,000 excess deaths globally are due to suspended particulate matter and about 370,000 to sulphur dioxide. PMID- 8711766 TI - Exposure to environmental chemicals relevant for respiratory hypersensitivity: European aspects. AB - The need for assessing air pollution exposure on the individual and population levels is urgent, as health effects are observed at concentrations even below existing air quality guidelines (AQG). Typical European concentrations and trends over time of four main air quality indicators (nitrogen dioxide, sulphur dioxide, ozone and particulate matter) are presented, together with their revised WHO AQG. In order to describe concentrations of pollutants each individual and population groups are exposed to, methods have been developed that estimate these concentrations using different forms of modelling. The EXPO model, developed to assess European exposure to air quality contaminants, estimates exceedances of AQG in many European cities and serves as an initial step in estimating European population exposure. Individual pollution exposure is estimated by measuring/estimating pollution concentrations at the home and/or by following and integrating exposure in different micro-environments. Examples of these methods are presented. PMID- 8711767 TI - Environmental chemicals relevant for respiratory hypersensitivity: the indoor environment. AB - The allergenic constituents of non-industrial indoor environments are predominantly found in the biologic fraction. Several reports have related biological particles such as mites and their excreta, dander from pets and other furred animals, fungi and bacteria to allergic manifestations including respiratory hypersensitivity among the occupants of buildings. Also, bacterial cell-wall components and the spores of toxin-producing moulds may contribute to the induction of hypersensitivity, but the relevance for human health is not yet determined. The knowledge regarding hypersensitivity and asthmatic reactions after exposure to chemical agents is primarily based on data from occupational settings with much higher exposure levels than usually found in non-industrial indoor environments. However, there is evidence that indoor exposure to tobacco smoke, some volatile organic compounds (VOC) and various combustion products (either by using unvented stoves or from outdoor sources) can be related to asthmatic symptoms. In some susceptible individuals, the development of respiratory hypersensitivity or elicitation of asthmatic symptoms may also be related to the indiscriminate use of different household products followed by exposure to compounds such as diisocyanates, organic acid anhydrides, formaldehyde, styrene and hydroquinone. At present, the contribution of the indoor environment both to the development of respiratory hypersensitivity and for triggering asthmatic symptoms is far from elucidated. PMID- 8711768 TI - Interaction of environmental chemicals with respiratory sensitization. AB - The acute effects of the inhalation of air polluting agents have been examined by many research groups in both animal models and human beings. For instance, it is evident that exposure to ozone has toxic effects and can lead to lung function disturbances. For this reason it is likely that individuals suffering from COPD or asthma are groups especially at risk with respect to the effects of ozone. The majority of studies dealing with effects of air pollutants on pulmonary allergy are restricted to IgE mediated allergy (type I allergy). Again for ozone, in animal models for type I allergy it has been demonstrated that exposure can affect the induction as well as the effector phase of this type of hyperimmune reaction (e.g. allergic asthma). Recently it has been demonstration in animal models that non-IgE mediated "asthma' T cells, and notably Th1 cells, may play a crucial role. In a murine model it was demonstrated that low molecular weight compounds can induce delayed type hypersensitivity-like reactions in the respiratory tract, and that these reactions are associated with the induction of airway hyperreactivity. Such compounds include toluene diisocyanate (TDI), to which immune responses can be readily mounted, and which can cause occupational asthma through its sensitizing capability, but to which IgE is only detected in a minority of patients suffering from TDI-associated asthma. Effects of air pollutants on Th1 responses in the respiratory tract have not been studied so far. We have demonstrated that ozone can inhibit resistance to a an intratracheal challenge with Listeria monocytogenes, indicating suppression of Th1 immune responses. In addition, we have shown that ozone exposure suppresses pulmonary delayed type hypersensitivity induced by small molecular weight compounds, as well as the tracheal hyperreactivity that is induced during the development of these immune responses, again supporting the hypothesis of suppression of Th1 responses by ozone exposure. These phenomena may be due to activation of Th2 cell dependent reactions that in turn lead to a downregulation of Th1 mediated immunity, or to a direct effect on Th1 cells or other cell types that are crucial for delayed type hypersensitivity and related airway hyperresponsiveness in this model. These data indicate that exposure to air pollutants may have differential consequences on different types of immune responses in the respiratory tract. PMID- 8711769 TI - Air pollutants and respiratory hypersensitivity. AB - Epidemiological evidence suggests that an increase in liquid petroleum derived pollutants is associated with exacerbation of allergic airway disease, and that the effects of pollution may occur 1-2 days later. Laboratory based studies have demonstrated that the pollutants responsible for the adverse effects on respiratory health include nitrogen dioxide (NO2), sulphur dioxide (SO2), ozone (O3) and respirable particulates (PM10). More recently, studies of asthmatic individuals exposed to O3, NO2 and a combination of NO2 and SO2 have indicated that these agents increase the airway responsiveness of these individuals to inhaled allergen, and that this effect may be maximal 24 h after exposure to the pollutants. Studies investigating the putative mechanisms underlying the effects of these pollutants suggest that exposure to these agents may lead to perturbation of the airway epithelium and release of pro-inflammatory mediators from the epithelial cells, which then influence the activity of inflammatory cells, such as eosinophils. PMID- 8711770 TI - Predictive testing for respiratory sensitisation. AB - A rat bioassay has been developed to provide an objective approach for the identification and classification of upper and lower respiratory tract irritants, with particular emphasis on the concentration-dependent induction and regression of lesions characteristic of asthma, such as persisting non-specific airway hyperreactivity, inflammation and ensuing mismatch of the ventilation-perfusion relationship. For the identification of respiratory allergy, the established guinea-pig bioassay has been further refined. Refinement focused on procedures making this animal model more robust to changes in study design. Attempts were made to allow differentiation of non-specific and specific bronchial hyperresponsiveness and to minimise the use of hapten-protein conjugates for elicitation of respiratory allergy. It appears that the combined assessment of specific pathologic features such as airway eosinophilia and the evaluation of several breathing parameters during hapten and acetylcholine bronchoprovocation challenge make it easier to distinguish effects caused by irritation and respiratory hypersensitivity. Findings support the conclusion that current guinea pig models require specific optimisation of sensitisation and challenge procedures for each chemical class tested. PMID- 8711771 TI - Structure-activity relationships and computer-assisted analysis of respiratory sensitization potential. AB - The mechanism(s) underlying respiratory sensitivity to chemicals is uncertain but is assumed to involve immunologic components with pharmacologic and neurologic involvement. Predictive testing would be valuable to prevent occurrence of hypersensitivity. Several in vitro and in vivo approaches have been used for predictive purposes. In vitro methods have included assessment of the ability of the chemical to undergo reaction with proteins. Computational methods have investigated the relationship between structure and electrophilic potential of chemical allergens. We have initiated a structure-activity evaluation of chemicals associated with elicitation of respiratory sensitization and have utilized a computer-based expert system, MultiCASE. A preliminary database of 39 active chemicals has been established from a literature search of clinical case reports and animal test results. Evaluation of the model has indicated structural alerts for activity which consist of structural fragments as well as physicochemical properties. Further development of the model and evaluation of findings should enable mechanistic insight into the process of respiratory sensitization and recognition of factors which distinguish respiratory sensitizers mechanistically from other chemical allergens such as contact sensitizing chemicals. PMID- 8711772 TI - Predictive testing for respiratory sensitization in the mouse. AB - Attempts to develop predictive test methods for the identification of chemical respiratory allergens have to date focused almost exclusively on the guinea pig. In recent years there has, however, been a growing interest in the mouse as a model for examination of sensitization potential. In this article two alternative approaches to the toxicological investigation of respiratory sensitization are described. Both are based on an understanding of the nature of immune responses induced in mice by chemical allergens. The mouse IgE test seeks to identify chemicals capable of causing allergic sensitization of the respiratory tract as a function of induced increases in the serum concentration of IgE. The second approach, cytokine fingerprinting, makes use of the observation that chemical allergens of different types provoke in mice qualitatively divergent immune responses characterized by discrete cytokine secretion profiles. PMID- 8711773 TI - Socioeconomic factors and the development of allergy. AB - Allergy has been associated with relative affluence for over a century, and more recently a substantially lower prevalence of sensitisation to common aeroallergens has been demonstrated in parts of Eastern Europe compared with Western and Northern Europe. One of the strongest risk factors for atopy is small sibship size. This applies in both affluent and less affluent families, and in both Western and Eastern Europe. The protective effect of older siblings is stronger than that of younger siblings, suggesting that family structure in early life is important. A unifying explanation for all these observations would be that allergic sensitisation can be prevented by infections acquired during early childhood. Direct evidence in support of this hypothesis is still awaited, but immunological mechanisms have been suggested. A reduction in infection offers a more coherent explanation of past and current trends in allergy prevalence than does an increase in exposure to environmental chemicals. PMID- 8711774 TI - Infections and the development of allergy. AB - The development of resistance or sensitivity to environmental antigens depends to a large extent on the nature of immunological memory which is generated during early antigen encounters in infancy and early childhood. The latter involves antigen-driven selection for specific Th-1-like versus Th-2-like memory cells within individual immune responses to inhaled allergens, a process which occurs in the regional lymph nodes (RLN) draining the conducting airways and which is regulated by a variety of cytokines produced by antigen-specific CD4+ and CD8+ T cells. In addition, this T-cell selection process can theoretically be influenced by infectious agents, at two distinct levels. Firstly, infections in the airway mucosa may mobilise and activate local tissue macrophages, which migrate to RLN and secrete Th-2-inhibitory cytokines such as IL-12 and IFN alpha, and also add to IFN gamma levels in the milieu via stimulation of NK cells. Secondly, microbial stimulation via gastrointestinal tract (GIT) commensals and pathogens is recognised as the principal trigger for postnatal maturation of overall immune competence in mammals. Recent studies indicate that the speed with which the immune system in human infants attains adult-equivalent competence postnatally is inversely related to 'risk' for primary allergic sensitisation to environmental antigens. Factors which affect qualitative/quantitative aspects of microbial colonisation of the GIT during early postnatal life may accordingly have unexpected downstream effects on seemingly unrelated processes such as development of T cell memory to allergens. PMID- 8711775 TI - Environmental chemicals and respiratory hypersensitization: a synopsis. PMID- 8711776 TI - Environmental chemicals and respiratory hypersensitization. Conclusions and recommendations. PMID- 8711777 TI - Environmental chemicals, respiratory hypersensitization and international chemical safety. AB - Allergic hypersensitization to a variety of chemicals, natural and synthetic, is a worldwide health problem. Respiratory tract hypersensitization is responsible for significant morbidity and, in some cases, mortality. An important step in managing and controlling health risks, such as allergic hypersensitization, is to identify the chemical hazard, define dose-effect and dose-response relationships, evaluate exposure, and characterize risk. In practical terms, the risk and safety assessment processes lead to the designation of control limits for exposure to chemicals in air, food, water, and consumer products. The objective of exposure limits is to protect the whole human population, including the most susceptible individuals and 'at risk' groups. The existence of susceptible individuals is a factor that must be taken into account when quantitative chemical risk assessments are being made, and should be covered in the risk characterization. Within a population, individual susceptibility is influenced by genetic and environmental factors and these have regional and national differences. There may be cases where hypersusceptible individuals and groups, such as asthmatic children, are not fully protected by regulatory exposure limits. The International Programme on Chemical Safety, as the global programme on identifying and assessing chemical risks to human health and the environment in order to assist countries in effective management, is striving to elucidate the toxicological basis for chemically-associated disease and advance the basic science and methodology of chemical risk and safety assessment. PMID- 8711778 TI - The immunology of respiratory allergies. AB - The main function of the respiratory tract is to provide a large surface area of thin epithelium for gas exchange. At the same time, this exposed surface and the conducting airways have to be defended against airborne irritants and infectious agents. The principal defence is the barrier formed by airway mucus and the mucociliary escalator. Agents which penetrate the initial defences may be destroyed by phagocytic cells, and may initiate an immune response. Respiratory allergy results when airborne allergens penetrate these defences and elicit and unhelpful immunological response. The nature of the airway immune response depends on the nature of the allergen, the antigen-processing pathway, and the microenvironment which dictates the phenotype of available T lymphocytes. Most allergens elicit IgE antibodies which then bind to mast cells and, when cross linked, the mast cell releases inflammatory mediators which cause bronchospasm and mucus formation. Some chemical allergens appear able to trigger this pathway without involving IgE. In both cases, other inflammatory cells, especially eosinophils, are then recruited. These cells appear to be responsible for the epithelial damage and increased airways reactivity that characterise asthma. Similar histological patterns are found in atopic asthma, non-atopic asthma, occupational asthma due to low molecular weight chemicals and even in the reactive airways dysfunction syndrome (RADS)/irritant-induced asthma syndrome. Allergic airway inflammation and clinical asthma appear to be common histological and clinical consequences of a variety of specific and non-specific insults to the airways epithelium, airways mast cells and airways T lymphocytes. PMID- 8711779 TI - Genetics of asthma and atopy. AB - Asthma and airways hyperresponsiveness are the respiratory manifestations of sensitization to exogenous materials including protein allergens and chemical sensitizers that may trigger ongoing inflammation and respiratory symptomatology in those people who are predisposed to develop the asthmatic syndrome. That genetics plays a major role in this particular syndrome is without question, since twin studies have shown a greater prevalence of disease for monozygotic as opposed to dizygotic twins. There have been the beginnings of the development of an approach to a number of candidate genes for major genetic input in the asthmatic syndrome. However, unlike cystic fibrosis or even other autoimmune and inflammatory diseases such as juvenile diabetes and/or multiple sclerosis, a single gene or a significant set of major genes may not be easily identified. Rather the small contribution of multiple genes and/or sets of genes may be summed together and added to environmental exposure in people who will develop asthmatic syndromes. The identification of a genetic component to asthma would be of great significance. Even without a major or predominant gene, identification of the minor sets of genes interacting to cause asthma would represent a great advance. Knowledge of those minor genetic alleles involved in asthma susceptibility would allow great latitude in offering diagnostic screening and new therapeutic interventions in asthma. PMID- 8711780 TI - Chemicals and proteins as allergens and adjuvants. AB - Small molecular weight chemicals may irritate tissues via the induction of the production of various proinflammatory and chemotactic molecules. The structure of these irritants is heterogenous, as is the pattern of their effects. Chemicals are potentially allergenic (i.e. haptens) when they are able to bind proteins such as immune response molecules. Sensitization will occur when these haptens additionally induce irritation resulting from an intrinsic adjuvant's activity of the chemical. In most cases haptens induce the activation of hapten-specific T cells with a type 1 cytokine profile mediating delayed hypersensitivity. A limited number of haptens induce the production of type 2 cytokines in T cells and, consequently, allergic reactions after inhalation. Similarly, inhaled proteins become allergenic when they activate protein allergen-specific T cells producing type 2 cytokines. In many individuals this cytokine profile is associated with atopy. It must be expected, however, that a type 2 cytokine profile can also be inflicted by the action of various adjuvants types, such as biologically active small molecular weight chemicals and proteins (i.e. enzymes) and microorganisms, thereby promoting allergic reactions. PMID- 8711781 TI - The role of the gastrointestinal tract in the development of respiratory hypersensitivities. AB - Adverse reactions to foods may sometimes cause symptoms from the respiratory tract, including bronchial obstruction and rhinitis. The true prevalence is not known. In adults, it has been estimated to be about 1% of asthmatic patients or less. This would correspond to a prevalence of about 2-4 in 10,000 of the general population. The prevalence in children is higher, as IgE mediated food allergy is more common in this age group than in adults. The most common foods causing immunologically mediated reactions include milk, egg, fish, crustaceans, nuts, wheat, soy, peanut, peas and other legumes. In addition certain food additives, e.g. sulphites, may rarely be incriminated in respiratory hypersensitivity via adverse reactions with unknown mechanisms. A double-blind placebo controlled food challenge (DBPCFC) is the only way to conclusively confirm a relationship between the ingestion of a certain food item and a reaction in the respiratory tract. PMID- 8711782 TI - The role of the skin in the development of chemical respiratory hypersensitivity. AB - Certain chemicals are known to cause occupational respiratory allergy associated with symptoms of pulmonary distress, including asthma and rhinitis. While there is no doubt that inhalation represents an important route of exposure for the development of sensitization to the inducing allergen, there is evidence that effective sensitization of the respiratory tract may result also from dermal contact with the chemical. The mechanisms relevant to the stimulation of respiratory sensitization following cutaneous exposure to chemical allergens and implications for the prevention of occupational asthma are considered here. PMID- 8711783 TI - Environmental factors and respiratory hypersensitivity: experiences from studies in Eastern and Western Europe. AB - Allergy and respiratory hypersensitivity are the end results of an interaction in genetically susceptible individuals, exposure to allergens and adjuvant factors. The incidence of allergic disease (but not respiratory hypersensitivity) is higher in urban than in rural areas and in Western industrialised countries than in the formerly socialist countries of Eastern Europe. It appears to be caused by factors encountered in early childhood. Tobacco smoke is by far the most important single environmental risk factor that has been identified so far. Other risk factors include air pollution such as NO2, SO2 and ozone, and modern, tightly isolated and poorly ventilated houses. However, even if all known environmental factors are added, this could only partly explain the increasing prevalence of these diseases. Therefore, other, unknown factors associated with 'Western lifestyle' must be looked for. PMID- 8711784 TI - The International Study of Asthma and Allergies in Childhood (ISAAC): objectives and methods; results from German ISAAC centres concerning traffic density and wheezing and allergic rhinitis. AB - The International Study of Asthma and Allergies in Childhood (ISAAC) was founded to maximize the value of epidemiological research into asthma and allergic disease by establishing a standardized methodology and facilitating international collaboration and comparison of data. The ISAAC design comprises three phases: Phase I uses validated core questionnaires designed to assess the prevalence and severity of asthma and allergic disease in defined populations; Phase II will investigate possible aetiological factors suggested by findings of Phase I and apply lung, blood and skin tests; Phase III will be a repetition of Phase I after 5 years to assess time trends in the prevalence and severity of wheezing, rhinitis and eczema in each ISAAC centre. In 1991 a survey on wheezing and allergic rhinitis in 2050 12-15 year old adolescents in Bochum showed a positive correlation between the prevalence of wheezing as well as allergic rhinitis and indicators of traffic density, controlling for putative confounders such as age, sex, passive smoking, active smoking, etc. In 1994-1995 an ISAAC Phase I survey conducted on adolescents in Munster, applying methodology similar to that in Bochum, found positive associations between surrogate measures of traffic density and 12 months prevalence of wheezing and lifetime prevalence of allergic rhinitis of a magnitude very close to that found in Bochum. The results from the two German cities provide support to the hypothesis that exposure to automobile emissions is related to wheezing and allergic rhinitis in children. PMID- 8711785 TI - From UBOs to Binswanger's disease. Impact of magnetic resonance imaging on vascular dementia research. PMID- 8711786 TI - Clinical correlates of white matter findings on cranial magnetic resonance imaging of 3301 elderly people. The Cardiovascular Health Study. AB - BACKGROUND AND PURPOSE: Our aim was to identify potential risk factors for and clinical manifestations of white matter findings on cranial MRI in elderly people. METHODS: Medicare eligibility lists were used to obtain a representative sample of 5888 community-dwelling people aged 65 years or older. Correlates of white matter findings were sought among 3301 participants who underwent MRI scanning and denied a history of stroke or transient ischemic attack. Participants underwent extensive standardized evaluations at baseline and on follow-up, including standard questionnaires, physical examination, multiple blood tests, electrocardiogram, pulmonary function tests, carotid sonography, and M-mode echocardiography. Neuroradiologists graded white matter findings from 0 (none) to 9 (maximal) without clinical information. RESULTS: Many potential risk factors were related to the white matter grade, but in the multivariate model the factors significantly (all P < .01) and independently associated with increased grade were greater age, clinically silent stroke on MRI, higher systolic blood pressure, lower forced expiratory volume in 1 second (FEV1), and income less than $50,000 per year. If excluded, FEV1 was replaced in the model by female sex, history of smoking, and history of physician-diagnosed hypertension at the baseline examination. Many clinical features were correlated with the white matter grade, especially those indicating impaired cognitive and lower extremity function. CONCLUSIONS: White matter findings were significantly associated with age, silent stroke, hypertension, FEV1, and income. The white matter findings may not be considered benign because they are associated with impaired cognitive and lower extremity function. PMID- 8711787 TI - Risk factors for incident dementia after stroke. Role of hypoxic and ischemic disorders. AB - BACKGROUND AND PURPOSE: Stroke significantly increases the risk of dementia in the elderly, yet the risk factors for incident dementia after ischemic stroke are not well understood. We attempted to determine whether hypoxic-ischemic (HI) disorders, which may result from comorbid medical conditions (eg. seizures, cardiac arrhythmias, pneumonia), would be an independent risk factor for the development of new dementia after stroke. METHODS: We prospectively followed 185 initially nondemented patients with ischemic stroke (age, 70.3 +/- 7.7 years) for a maximum of 52.8 months. We diagnosed the presence of dementia at annual examinations based on neuropsychological testing and modified DSM-III-R criteria. HI disorders were identified by record review or examination during hospitalization. We used Kaplan-Meier analysis to determine the cumulative proportion of patients with and without HI disorders who survived free of dementia and used Cox models to estimate the relative risk of dementia associated with HI disorders. RESULTS: The cumulative proportion (+/- SE) surviving without dementia was 51.7 +/- 10.9% in the HI group versus 78.2 +/- 4.3% in the non-HI group after 52.8 months of observation. The relative risk of incident dementia associated with HI events was 4.3 (95% confidence interval = 1.9 to 9.6) after we adjusted for demographic factors, recurrent stroke, and baseline cognitive function. CONCLUSIONS: We conclude that HI disorders may be a significant independent risk factor for incident dementia after stroke, even after adjustment for other recognized predictors of cognitive decline. Recognition of HI cerebral damage as a possible pathogenic mechanism for dementia after stroke may allow targeted therapeutic interventions to prevent subsequent cognitive deterioration. PMID- 8711788 TI - Ultrasonographic assessment of carotid wall characteristics and cognitive functions in a community sample of 59- to 71-year-olds. The EVA Study Group. AB - BACKGROUND AND PURPOSE: This study was aimed at analyzing cross-sectional relationships between cognitive performance and ultrasonographic assessment of carotid wall characteristics. METHODS: A cohort of 1279 subjects (men, 41%) aged 59 to 71 years was recruited from the electoral rolls of the city of Nantes (western France). Cognitive performances were evaluated with the Mini-Mental State Examination (MMSE) and seven neuropsychological tests assessing attention, psychomotor rapidity, verbal abilities, memory, and visuospatial perception. For each test, subjects were classified into three performance levels with a quartile distribution: 25% highest, 25% lowest, and 50% middle. The intima-media thickness of common carotid arteries and the presence of plaques in the carotid arteries were assessed with B-mode ultrasound examination. RESULTS: Only 28% of men and 17% of women had carotid plaques inducing moderate stenosis of the lumen ( < 40%). After adjustment for possible confounders, odds ratios for poor cognitive performance associated with plaques were above 1 for all cognitive tests in men. This association was statistically significant for the MMSE and another test assessing attention skills. There was a slight association between increase of the common carotid intima-media thickness and poor cognitive scores in men with plaques. In women, no association was found between cognitive functions and presence of plaques or intima-media thickness. CONCLUSIONS: This study indicated a moderate association between atherosclerosis of the carotid arteries and poor cognitive functioning in men aged 59 to 71 years. In view of these moderate cross sectional results, further studies are required to better assess the relationship between carotid atherosclerosis and cognitive impairment. PMID- 8711789 TI - Hemostatic markers in acute ischemic stroke. Association with stroke type, severity, and outcome. AB - BACKGROUND AND PURPOSE: Hemostatic markers can identify activation of the coagulation system in stroke patients. We evaluated whether the levels of these markers at the time of stroke are correlated with stroke severity, type, or mortality. METHODS: We measured fibrinopeptide A, cross-linked D-dimer, and beta thromboglobulin in 70 patients within 1 week of stroke. We examined the association between the level of each of these markers and survival. We adjusted for the possible confounding effect of age, stroke type, or stroke severity using a multivariate Cox proportional hazards model. RESULTS: The median follow-up was 1.22 years. Fourteen patients died during follow-up. Univariate survival analysis identified age (hazard ratio, 1.06; 95% confidence interval [CI], 1.00 to 1.12), stroke type (hazard ratio, 4.44; 95% CI, 1.29 to 15.23), initial Toronto Stroke Scale score (hazard ratio, 5.05; 95% CI, 2.08 to 12.27), cross-linked D-dimer (hazard ratio, 6.43; 95% CI, 2.83 to 14.62), fibrinopeptide A (hazard ratio, 2.14; 95% CI, 1.26 to 3.63), and beta-thromboglobulin (hazard ratio, 7.63; 95% CI, 2.22 to 26.28) as significantly associated with mortality. In a multivariate model, initial stroke severity and each of the hemostatic markers were independently associated with subsequent mortality. CONCLUSIONS: Elevated hemostatic markers after acute ischemic stroke identify patients with increased risk for mortality. This association appears to be independent of stroke severity or stroke type. PMID- 8711790 TI - Outcome in patients with acute basilar artery occlusion requiring mechanical ventilation. AB - BACKGROUND AND PURPOSE: Many patients with acute basilar artery occlusion may require endotracheal intubation and mechanical ventilation. The circumstances and predictive value for outcome in these patients are not well documented. METHODS: We reviewed the medical records of 25 patients admitted into the intensive care unit with a clinical diagnosis of acute basilar artery occlusion and need for mechanical ventilation. The medical records were reviewed for clinical features, breathing patterns, mode of mechanical ventilation, ability to wean from the ventilator, and neurological outcome. RESULTS: Apneic episodes resulted in endotracheal intubation in 8 patients. In the remaining 17 patients, intubation was needed for airway protection. Seven of 8 patients presenting with apneic episodes lost all brain stem reflexes. All 17 patients intubated for airway protection could be successfully weaned to a T-tube circuit. Outcome was generally poor and 22 patients died, of whom 7 died of early systemic complications. Only 3 of 25 patients, all with locked-in syndrome, survived. CONCLUSIONS: Mortality is high in patients who require mechanical ventilation after acute basilar artery occlusion. No neurological improvement beyond a locked in syndrome occurred in survivors. Recurrent apnea appears to predict further progression to brain stem death. PMID- 8711791 TI - The ABCs of measuring intracerebral hemorrhage volumes. AB - BACKGROUND AND PURPOSE: Hemorrhage volume is a powerful predictor of 30-day mortality after spontaneous intracerebral hemorrhage (ICH). We compared a bedside method of measuring CT ICH volume with measurements made by computer-assisted planimetric image analysis. METHODS: The formula ABC/2 was used, where A is the greatest hemorrhage diameter by CT, B is the diameter 90 degrees to A, and C is the approximate number of CT slices with hemorrhage multiplied by the slice thickness. RESULTS: The ICH volumes for 118 patients were evaluated in a mean of 38 seconds and correlated with planimetric measurements (R2 = 9.6). Interrater and intrarater reliability were excellent, with an intraclass correlation of .99 for both. CONCLUSIONS: We conclude that ICH volume can be accurately estimated in less than 1 minute with the simple formula ABC/2. PMID- 8711792 TI - Posterior circulation infarcts simulating anterior circulation stroke. Perspective of the acute phase. AB - BACKGROUND AND PURPOSE: Ischemic stroke patients whose initial clinical presentation suggests an involvement of the anterior circulation (AC) are sometimes found to have a posterior circulation (PC) infarct, a fact that may generate erroneous decisions in clinical management. We investigated the prevalence of this misdiagnosis in the first few hours after stroke onset. METHODS: We performed a cohort study of 158 patients hospitalized within 5 hours of onset of a presumed AC ischemic stroke, as diagnosed on clinical grounds. RESULTS: Final CT or pathology diagnosis was AC infarct in 128 patients (81%), a repeatedly negative CT in 14 (9%), PC infarct (5 pons, 1 midbrain and cerebellum, 6 supratentorial territory of the posterior cerebral artery) in 12 (8%), and other or undiagnosed lesions in 4 (3%). AC and PC stroke patients did not differ in terms of age, vascular risk factors, and initial severity, but the latter were more frequently men (83% versus 53%; P = .04), were hospitalized later (mean +/- SD, 168 +/- 86 versus 109 +/- 55 minutes; P = .001), and presented a pure motor hemiparesis or a sensorimotor stroke (50% versus 33%) more often than their counterparts. At baseline CT, PC stroke patients never exhibited an early parenchymal hypodensity in the carotid territory or a hyperdense middle cerebral artery, which were instead found in 59% (P = .0003) and 31% (P = .02) of AC stroke patients, respectively. Early neurological deterioration, 1 month case fatality rate, and disablement in survivors were comparable in the two groups. CONCLUSIONS: Shortly after onset the clinical discrimination between AC and nontypical PC infarcts is not reliable, which explains the frequent occurrence of this misdiagnosis. Emergency CT scan helps in the differential diagnosis only when it demonstrates an early focal hypodensity within the carotid territory. PMID- 8711793 TI - Apolipoprotein E polymorphism and stroke in a population sample aged 75 years or more. AB - BACKGROUND AND PURPOSE: We investigated apolipoprotein E polymorphism stroke risk in a population sample of 1810 persons aged 75 years or more in Stockholm (the Kungsholmen Project). Information on cognition at cohort inception (from 1987 to 1989) and on stroke occurrence (from 1969 to 1994) is available for the cohort. In the cohort, cognitive impairment is associated with the epsilon 4 allele, and longer survival in subjects aged > or = 85 years with good cognition is associated with the epsilon 2 allele and the absence of epsilon 4. METHODS: We compared stroke incidence in the 1077 of 1124 genotyped subjects who carried epsilon 2/3, epsilon 3/3, or epsilon 3/4 and estimated the proportion of cognitive impairment attributable to stroke. RESULTS: Risk of stroke did not vary with apolipoprotein E polymorphism (P = .82): 24% of 87 incident stroke patients during follow-up compared with 25% of 827 subjects with normal cognition and no stroke diagnosis at baseline carried the epsilon 3/4 genotype. An estimated 9% of cognitive impairment was attributable to stroke. Notably, a reduced epsilon 3/4 frequency of 20% was found in subjects who survived a prior stroke and were included in the cohort, and risk of hemorrhagic stroke tended to be associated with the presence of the epsilon 3/4 genotype and the absence of epsilon 2/3. CONCLUSIONS: This population-based study indicates that apolipoprotein E polymorphism is not a risk factor for ischemic stroke in subjects aged > or = 75 years (although it might possibly influence survival after stroke occurrence and be a risk factor for infrequent hemorrhagic stroke) and that approximately 10% of cognitive impairment in this age group is attributable to stroke. PMID- 8711794 TI - Autonomic neuropathy predicts the development of stroke in patients with non insulin-dependent diabetes mellitus. AB - BACKGROUND AND PURPOSE: Our aim was to determine the predictive factors for stroke in patients with non-insulin-dependent diabetes mellitus (NIDDM). METHODS: We studied 133 patients with NIDDM at the time of diagnosis and 5 and 10 years later. RESULTS: The number of new fatal or nonfatal strokes was 19 (14.7%; 14 after 5-year examination). High initial fasting blood glucose (odds ratio [OR], 1.2; 95% confidence interval [CI], 1.04 to 1.4) and the use of beta-blocking agents (OR, 6.7; 95% CI, 2.1 to 21.5) at baseline and the presence of parasympathetic neuropathy (OR, 6.7; 95% CI, 1.5 to 29.9), or sympathetic autonomic nervous dysfunction (OR, 1.1; 95% CI, 1.01 to 1.2), hypertriglyceridemia (OR, 5.7; 95% CI, 1.1 to 31.0), or use of beta-blocking agents (OR, 6.4; 95% CI, 1.3 to 31.2), and high fasting plasma glucose (OR, 1.2; 95% CI, 1.0 to 1.5) determined at 5-year examination predicted the development of stroke. CONCLUSIONS: Autonomic neuropathy is an independent risk factor for stroke in NIDDM. PMID- 8711795 TI - Nocturnal blood pressure and silent cerebrovascular lesions in elderly Japanese. AB - BACKGROUND AND PURPOSE: We conducted a cross-sectional epidemiological survey using ambulatory blood pressure monitoring and brain MRI in a cohort from northern Japan to determine whether an inappropriately low nocturnal blood pressure, or an excess fall in nocturnal blood pressure, might be responsible for silent cerebrovascular lesions in the elderly. METHODS: Untreated subjects over 55 years and under 64 years of age (late middle age; 24 men and 46 women, 60% of eligible people) and over 65 years and under 75 years of age (elderly; 29 men and 52 women, 91% of eligible people) participated in the study. We evaluated the relationship between the amplitude (Daytime Average-Nighttime Average) or the rate ([Daytime Average-Nighttime Average]/Daytime Average) of the fall in nocturnal blood pressure and the incidence of silent cerebrovascular lesions on MRI (number of lacunar infarctions or extent of periventricular hyperintensity). RESULTS: The amplitude or the rate of the fall in nocturnal blood pressure in elderly women with one or two lacunar infarctions was significantly higher than that in those without such infarctions. There was a significant positive correlation between the amplitude or the rate of the fall in nocturnal blood pressure and the extent of periventricular hyperintensity in the elderly women. This relationship was observed in women, but not in men, of late middle age; this was not seen in elderly men. CONCLUSIONS: Results indicate that an inappropriately low nocturnal blood pressure, or an excessive fall in nocturnal blood pressure, is associated with ischemic silent cerebrovascular lesions, at least in elderly women. Treatment of hypertension in such women should be administered with care and with regard to nocturnal blood pressure. PMID- 8711796 TI - Effects of defibrination on hemorheology, cerebral blood flow velocity, and CO2 reactivity during hypocapnia in normal subjects. AB - BACKGROUND AND PURPOSE: Plasma fibrinogen is reported to be an independent risk factor for stroke and cardiovascular diseases. The effects of defibrination on hemorheology, middle cerebral artery (MCA) blood flow velocity, and CO2 reactivity during hypocapnia were evaluated in normal subjects. METHODS: Twenty five healthy subjects (mean age, 31.8 +/- 5.7 years) were included in the study. Measurements were done at rest and repeated 24 hours after administration of 10 batroxobin units. Plasma fibrinogen, plasma viscosity, and whole blood viscosity were measured as hemorheological factors. MCA blood flow velocity was measured with a transcranial Doppler flowmeter. Blood flow velocity was corrected to 40 mm Hg of end-tidal CO2 partial pressure (PETCO2), and expressed as CV40. CO2 reactivity was measured as percent change in mean blood flow velocity per millimeter of mercury PETCO2. RESULTS: Plasma fibrinogen (from 7.04 to 2.29 mumol/L; P < .001), whole blood viscosity, and plasma viscosity decreased after administration of batroxobin. Mean MCA blood flow velocity at rest, CV40. and CO2 reactivity during hypocapnia increased significantly (from 67.4 to 73.6 cm/s, from 71.7 to 77.7 cm/s, and from 2.9%/mm Hg to 3.2%/mm Hg, respectively; P < .01) after defibrination. Mean arterial blood pressure and PETCO2 at rest were constant before and 24 hours after administration of batroxobin. There was a significant positive correlation between CV40 and CO2 reactivity (r = .623, P < .0001). CONCLUSIONS: The increase in MCA blood flow velocity was associated with improved CO2 reactivity and reduced blood viscosity after defibrination. The data may suggest that defibrination increases cerebral blood flow by reducing blood viscosity. PMID- 8711797 TI - Apolipoprotein E epsilon 4 is associated with the presence and earlier onset of hemorrhage in cerebral amyloid angiopathy. AB - BACKGROUND AND PURPOSE: Cerebral amyloid angiopathy is an important cause of intracerebral hemorrhage in the elderly. The epsilon 4 allele of the apolipoprotein E gene, recently established as a genetic risk for Alzheimer's disease, has also been suggested as a possible risk factor for cerebral amyloid angiopathy. We sought to determine whether this allele is specifically associated with hemorrhages related to amyloid angiopathy and whether it correlates with the age at which first amyloid angiopathy-related hemorrhage occurs. METHODS: Forty five consecutive patients presenting with lobar hemorrhage were prospectively classified according to clinical, radiological, and when available, pathological features and evaluated for apolipoprotein E genotype. They were compared with 1899 elderly patients from a population-based sample and with 18 consecutive patients with hemorrhages in deep regions typical of a hypertensive mechanism. RESULTS: Patients with multiple hemorrhages confined to the lobar territory demonstrated a greater than twofold overrepresentation (P < .001) in frequency of the apolipoprotein E epsilon 4 allele compared with the population-based sample. Apolipoprotein E genotypes of patients with hemorrhages in deep territories resembled the population sample. Among patients with strictly lobar hemorrhages, carriers of the epsilon 4 allele had their first hemorrhage more than 5 years earlier than noncarriers (mean age at first hemorrhage, 73.4 +/- 8.0 versus 78.9 +/- 7.4 years; P = .033). These effects were independent of the accompanying presence of Alzheimer's disease. CONCLUSIONS: The data support a specific role for apolipoprotein E epsilon 4 in accelerating the process that leads to amyloid angiopathy-related hemorrhage. PMID- 8711798 TI - Prevalence of stroke in Kinmen. AB - BACKGROUND AND PURPOSE: Stroke is the second most common cause of death in Taiwan. We studied its prevalence, risk factors, and mortality in a rural Chinese population. METHODS: A door-to-door survey of stroke was conducted in two Kinmen Island townships with a total population of 26105 people. Our target population (n = 5061) consisted of all the registered residents in these townships who were aged > or = 50 years on August 1, 1993. All participants were given a standardized neurological examination and a questionnaire. RESULTS: The participation rate was 77.4% (n = 3915). Ninety-six cases of completed stroke were identified. Eighty-nine patients had one, 6 patients had two, and 1 patient had three episodes of stroke. The prevalence of stroke in persons aged > or = 50 years was 24.5 per 1000 (95% confidence interval, 19.7 to 29.3 per 1000). Prevalence increased with age. Statistically significant risk factors associated with stroke included hypertension, diabetes mellitus, and irregular heartbeats. Of the stroke survivors, 59% were independent in activities of daily living and 71% could walk independently. CONCLUSIONS: Compared with other countries, Taiwan has a moderately high prevalence of stroke. The risk factors for stroke in this rural region of a developing country are similar to those in developed countries. Most stroke survivors here perform their daily activities independently without outside assistance. PMID- 8711799 TI - Surgical therapy for adult moyamoya disease. Can surgical revascularization prevent the recurrence of intracerebral hemorrhage? AB - BACKGROUND AND PURPOSE: It is well recognized that revascularization surgery using direct and/or indirect bypass provides effective surgical management for pediatric moyamoya disease. However, surgical treatment of the adult hemorrhagic type remains controversial. In this study, the effect of surgery for adult moyamoya disease was investigated. METHODS: We analyzed 35 patients with adult moyamoya disease (patient age, over 20 years), 24 patients with initial onset of intracerebral hemorrhage, and 11 patients with initial onset of cerebral ischemia who underwent both direct bypass surgery of the superficial temporal artery to the middle cerebral artery anastomosis and indirect revascularization of encephalo-duro-arteriomyo-synangiosis. RESULTS: Of 24 patients with hemorrhagic type disease, 3 showed rebleeding: of 11 patients with the ischemic type, 2 showed intracerebral hemorrhage after surgery. Overall, 5 of 35 patients (14.3%) had hemorrhage after revascularization surgery (mean follow-up period, 6.4 years). Postoperative angiography revealed that direct anastomosis is effective whereas indirect revascularization is not always effective for adult moyamoya disease. Moyamoya vessels, which are supposed to be responsible for hemorrhage, decreased in 25% of patients. CONCLUSIONS: Revascularization surgery cannot always prevent rebleeding. However, a decrease in moyamoya vessels was induced by surgery, which may reduce the risk of hemorrhage more effectively than conservative treatment. In cases of adult moyamoya disease, direct bypass is particularly important, since the indirect revascularization is not as useful in adult cases as in pediatric cases. PMID- 8711800 TI - Detection of microembolic signals in patients with middle cerebral artery stenosis by means of a bigate probe. A pilot study. AB - BACKGROUND AND PURPOSE: Middle cerebral artery (MCA) stenosis is a relatively rare occlusive disease with an annual stroke risk of approximately 7% to 8%. However, the frequent coincidence of cardiac or ipsilateral carotid artery disease may lead to difficulties in identifying the relevant embolizing source in symptomatic patients. We undertook this study to evaluate the prevalence of microembolic signals (MES) as well as the potential and limitations of bigate monitoring in patients with MCA stenosis. METHODS: Fourteen patients aged 33 to 87 years with angiographically demonstrated symptomatic (acute, n = 2; chronic, n = 8) or asymptomatic (n = 4) MCA stenosis were examined. Six patients (43%) had additional cardiac (n = 3) or carotid artery (n = 3) disease. By means of a bigate probe, simultaneous insonation of prestenotic and poststenotic vessel segments was attempted. RESULTS: In 10 patients (71%), MES detection could be performed sufficiently at target vessel sites. In the remaining patients, either prestenotic (n = 3) or poststenotic (n = 1) monitoring was not satisfactory due to insufficient transtemporal bone window or the great length or extent of MCA stenosis. Poststenotic MES were detectable in 2 acutely symptomatic and 1 asymptomatic patient (prevalence, 21%). In the latter case, the sequential appearance of MES in both prestenotic and poststenotic channels excluded MCA stenosis but strongly favored coexisting carotid artery stenosis as the active embolic source. CONCLUSIONS: MES are detectable in patients with MCA stenosis. Bigate monitoring in this setting is feasible and allows identification of the active source among "competing" embolizing conditions. PMID- 8711801 TI - A transcranial doppler ultrasonography study of cerebrovascular CO2 reactivity in mitochondrial encephalomyopathy. AB - BACKGROUND AND PURPOSE: To elucidate the pathogenic role of vascular involvement such as mitochondrial angiopathy in patients with mitochondrial encephalomyopathy (MEM). we used the transcranial Doppler sonography (TCD) method to detect impairment of cerebrovascular CO2 reactivity. METHODS: The cerebral perfusion reserve in 13 MEM patients, including 6 with MELAS (mitochondrial myopathy, encephalopathy, lactic acidosis, and strokelike episodes) was studied by TCD for different CO2 partial pressures. For the parameter of mean flow velocity, the mean spatial Doppler frequency (fm) was obtained from the right and left middle cerebral arteries and basilar artery under conditions of normocapnia, hypercapnia, and hypocapnia in cases in which hyperventilation was possible. By fitting the obtained fm and the end-tidal CO2 partial pressure (PETCO2) to the exponential formula fm = a x e(K < PETCO2), where a is the theoretical fm at a PETCO2 of 0 mm Hg, the parameter K, an index of CO2 reactivity, was calculated. RESULTS: The K value was lower than control values at at least one site of the middle cerebral arteries and basilar artery of all patients with MELAS as well as the other MEM patients except for one patient with myoclonic epilepsy with ragged red fiber and one with Kearns-Sayer syndrome. CONCLUSIONS: Our results suggest that there is a high incidence of impairment of cerebrovascular CO2 reactivity in MEM patients. Moreover, the noninvasive TCD method was found useful for evaluation of cerebral hemodynamics in MEM patients. PMID- 8711802 TI - Angular biofeedback device for sitting balance of stroke patients. AB - BACKGROUND AND PURPOSE: Impaired sitting balance is an important and time consuming complication for stroke patients. We examined the effect of the use of an angular biofeedback device in addition to physical therapy in training stroke patients with impaired sitting balance compared with outcome in patients receiving conventional physical therapy only. METHODS: The biofeedback group consisted of 24 patients who received angular biofeedback training in addition to conventional physical therapy. The number of biofeedback signals and the lengths of time a patient could sit balanced throughout a period of 5 minutes before the training program, after 10 days of treatment, and at discharge were recorded and compared with those of the control group of 13 patients who received conventional physical therapy only. RESULTS: It was found that 75% of the biofeedback group gained sitting balance after 10 days of treatment in comparison with 15.4% of the control group (P < .001). At discharge, 91.6% of the biofeedback group and 84.6% of the control group gained sitting balance (P = .510), and 45.8% of the biofeedback group and 46.2% of the control group managed independent ambulation (P = .985). The mean rehabilitation periods among the ambulatory patients of the biofeedback and control groups were 9.45 +/- 0.71 and 13.83 +/- 1.70 weeks, respectively (P = .049). The mean training time in which the biofeedback group gained sitting balance was significantly shorter than that of the control group (P < .001). CONCLUSIONS: Angular biofeedback intervention, by providing earlier postural trunk control, is a useful adjunct to conventional physical therapy in the rehabilitation of stroke patients with impaired sitting balance. PMID- 8711803 TI - Transvenous hemodynamic assessment of arteriovenous malformations and fistulas. Preliminary clinical experience in Doppler guidewire monitoring of embolotherapy. AB - BACKGROUND AND PURPOSE: Transvenous monitoring of blood flow through intracranial vascular malformations was performed with an intravascular Doppler guidewire to assess hemodynamic changes during endovascular embolotherapy. METHODS: Flow velocity was assessed in the intracranial venous sinuses of two patients with arteriovenous malformations and seven patients with dural arteriovenous fistulas. In all cases, the Doppler guidewire was positioned in the dural sinuses coaxially through a 2.1F microcatheter. The Doppler guidewire was then advanced to the site of arteriovenous shunting for sampling of venous average peak velocity (APV) and pulsatility index. In two cases, simultaneous feeding artery flow velocity was monitored by transcranial color-coded duplex sonography. RESULTS: Before embolotherapy, the flow pattern in the venous sinuses was pulsatile, with a mean (+/-SD) APV of 39.0 +/- 22.5 cm/s. Total or near-total embolization was achieved in six of the nine cases. After embolization, the flow pattern became less pulsatile and the APV was reduced to a mean of 21.2 +/- 14.6 cm/s (P = .0123, one tailed paired t test). The pulsatility index was used to calculate the maximum minus the minimum peak velocity (MxPV-MnPV). This was reduced from an average of 27.0 +/- 8.7 cm/s to 13.5 +/- 8.3 cm/s after treatment (P = .0456). A parallel reduction in APV of the feeding arteries was observed with embolization. CONCLUSIONS: Preliminary clinical experience indicates that transvenous assessment of two parameters, APV and MxPV-MnPV, is useful in the hemodynamic evaluation of intracranial arteriovenous shunts. This valuable hemodynamic information may be used for objective and quantitative monitoring during embolotherapy of intracranial vascular malformations. PMID- 8711804 TI - Transvenous hemodynamic assessment of experimental arteriovenous malformations. Doppler guidewire monitoring of embolotherapy in a swine model. AB - BACKGROUND AND PURPOSE: A Doppler guidewire was used to monitor progressive changes in draining vein flow parameters during experimental embolotherapy in a swine arteriovenous malformation (AVM) model. METHODS: A microcatheter was positioned superselectively in the main arterial feeder and main draining vein in each of 10 AVM models in swine. With use of the Doppler guidewire, preembolization arterial and venous average peak velocities (APVs) and pulsatility indices were recorded. The device was left in the draining vein during transarterial particulate (in 8 swine) or liquid adhesive (in 2 swine) embolization, and continuous transvenous flow during and after treatment was monitored. Periembolization Doppler flow parameters were correlated qualitatively with angiographic changes in the nidus. RESULTS: Preembolization draining vein flow was pulsatile, with a mean APV of 38.9 +/- 13.7 cm/s. After embolization, this changed significantly to a less pulsatile or nonpulsatile pattern, with a lower mean APV of 9.2 +/- 4.9 cm/s (P = .0001). A novel expression, the maximum minus the minimum peak velocity (MxPV-MnPV), was used in evaluating the transvenous Doppler spectra. This was reduced significantly after embolization from a mean of 11.1 +/- 3.5 cm/s to 6.7 +/- 2.5 cm/s (P = .0025). Objective periembolization hemodynamic changes were detected in the draining veins earlier than the visually subjective angiographic changes within the nidus. CONCLUSIONS: Transvenous Doppler guidewire assessment of two parameters, APV and MxPV-MnPV, is useful in the hemodynamic evaluation of experimental arteriovenous shunting and may be used for future objective and quantitative monitoring during endovascular AVM embolotherapy. PMID- 8711805 TI - Inducible nitric oxide synthase gene expression in vascular cells after transient focal cerebral ischemia. AB - BACKGROUND AND PURPOSE: We investigated whether inducible nitric oxide synthase (iNOS) is expressed after transient cerebral ischemia and, if so, we sought to define the temporal profile and cellular localization of the expression and the role of iNOS in the mechanism of ischemic brain injury. METHODS: The middle cerebral artery in rats was occluded for 2 hours by an intraluminal filament. The occurrence of transient ischemia and reperfusion was confirmed by laser-Doppler flowmetry (n = 5). iNOS message in the ischemic neocortex was determined by reverse-transcription polymerase chain reaction. iNOS enzymatic activity was assessed by citrulline assay. The cellular localization of iNOS expression was determined by immunohistochemistry. RESULTS: iNOS mRNA was maximally expressed in postischemic brain at 12 hours and was not present at 4 days (n = 3 per time point). iNOS mRNA was not observed in the contralateral cerebral cortex. iNOS enzymatic activity developed in the postischemic brain between 12 and 24 hours (P < .05) and subsided at 4 days (n = 4 to 8 per time point). iNOS immunoreactivity in the ischemic region was restricted to the wall of capillaries and of larger blood vessels at 12 to 24 hours. In regions of early necrosis, inflammatory cells were iNOS positive. Treatment with the iNOS inhibitor aminoguanidine (n = 5; 100 mg/kg IP, BID for 4 days), starting 6 hours after ischemia, reduced infarct size in neocortex by 36 +/- 7% in comparison with vehicle-treated controls (n = 5) (P < .05). CONCLUSIONS: Transient focal ischemia leads to iNOS expression in postischemic brain. However, the spatial and temporal patterns of expression differ from those occurring in permanent ischemia: iNOS is induced earlier and predominantly in vascular cells rather than in neutrophils. Thus, the temporal profile and localization of postischemic iNOS expression depend on the nature of the ischemic insult. The finding that aminoguanidine reduces infarct size adds further support to the hypothesis that postischemic iNOS expression contributes to ischemic brain damage. PMID- 8711806 TI - Temporal correlation mapping analysis of the hemodynamic penumbra in mutant mice deficient in endothelial nitric oxide synthase gene expression. AB - BACKGROUND AND PURPOSE: Mice containing deletions in the genes encoding nitric oxide (NO) synthase have been useful to dissect the role of NO in cerebral ischemia. We recently reported that mice lacking expression of the endothelial isoform of NO synthase (eNOS) develop larger infarcts after middle cerebral artery occlusion. Because NO or a related product of NO synthase activity is important for relaxation of cerebral blood vessels, we examined for possible hemodynamic differences in the peri-ischemic zone of eNOS-deficient and wild-type mice after middle cerebral artery occlusion using functional CT scanning techniques. METHODS: Wild-type SV129 mice (n = 10) and mice deficient in eNOS gene expression (n = 10) were subjected to middle cerebral artery occlusion under halothane anesthesia. Thirty minutes after ischemia, functional CT scanning was performed with dynamic scanning protocols to measure the cerebral transit profiles of injected contrast agents. A temporal correlation mapping technique was used to analyze the pattern of hemodynamic perturbations based on alterations in the shape of the cerebral transit profiles. Statistical thresholds defined the hemodynamic core and penumbra. RESULTS: Hemodynamic deficits were more severe in the mutant than wild-type mouse. When expressed as a percentage of the total insult, core areas were significantly increased in mutant mice (39.8 +/- 3.7%) compared with wild types (28.8 +/- 3.4%). Conversely, areas of the hemodynamic penumbra were significantly smaller in mice deficient in eNOS activity (60.2 +/- 3.7%) than in wild-type mice (71.2 +/- 3.4%). Furthermore, the calculated relative perfusion index within the hemodynamic penumbra was significantly lower in the group with eNOS gene deletion (35.6 +/- 1.5% in mutants versus 43.0 +/- 2.4% in wild types). CONCLUSIONS: These data indicate that mice lacking eNOS expression show a greater degree of hemodynamic compromise after middle cerebral artery occlusion and suggest that a product of eNOS activity (eg. NO) may protect brain after focal cerebral ischemia, possibly by improving blood flow within the penumbral zone. PMID- 8711807 TI - E-selectin appears in nonischemic tissue during experimental focal cerebral ischemia. AB - BACKGROUND AND PURPOSE: E-selectin participates in leukocyte-endothelial adhesion and the inflammatory processes that follow focal cerebral ischemia and reperfusion. The temporal and topographical patterns of microvascular E-selectin presentation after experimental focal cerebral ischemia are relevant to microvascular reactivity to ischemia. METHODS: The upregulation and fate of E selectin antigen during 2 hours of middle cerebral artery occlusion (n = 4) and 3 hours of occlusion with reperfusion (1 hour, n = 4; 4 hours, n = 6; 24 hours, n = 6) were evaluated in the nonhuman primate. E-selectin and E:P-selectin immunoreactivities were semiquantitated with the use of computerized light microscopy video imaging and laser confocal microscopy. RESULTS: Three patterns of microvascular E-selectin expression, defined by the antibody E-1E4, were confirmed by complete elimination of E-1E4 binding after incubation with soluble recombinant human E-selectin: (1) Low immunoperoxidase intensity was observed in ischemic microvessels at 2 hours of occlusion extending to 4 hours of reperfusion (E-selectin/laminin = 0.32 +/- 0.10). (2) A significant fraction of ischemic microvessels displayed high-intensity E-selectin signal by 24 hours of reperfusion (0.61 +/- 0.17) compared with control and nonischemic tissues (2P < .003). (3) In the contralateral nonischemic basal ganglia and other nonischemic tissues, low but significant E-selectin levels appeared by 24 hours of reperfusion (2P = .0005). The latter were further confirmed by an E:P-selectin immunoprobe. CONCLUSIONS: E-selectin antigen is distinctively and significantly upregulated in nonhuman primate brain after focal ischemia and reperfusion. The late appearance of E-selectin in nonischemic cerebral tissues suggests stimulation by transferable factors generated during brain injury. PMID- 8711809 TI - Impaired autoregulation in an experimental model of chronic cerebral hypoperfusion in rats. AB - BACKGROUND AND PURPOSE: To verify the hypothesis that impaired autoregulation may contribute to cerebral swelling or hemorrhage after a sudden recovery of perfusion pressure, we studied the chronic effects of cerebral hypoperfusion on the autoregulatory responses of the pial arterioles in situ. METHODS: Eight to 12 weeks after a carotid-jugular fistula was created in rats, experiments were performed under alpha-chloralose and urethane anesthesia. Regional cerebral blood flow (rCBF) was determined by the hydrogen clearance method, and carotid pressure was measured. Using a closed cranial window, we determined the autoregulatory responses of the arterioles (30 to 50 microns) to both hypertension induced by norepinephrine and sudden fistula closure at various mean arterial pressures (MAPs). RESULTS: rCBF on the fistula side was reduced by 27%. Carotid pressure was significantly lower than normal but was immediately increased by fistula closure. The pial arterioles showed marked elongation and enlargement. During induced hypertension, the arterioles in the fistula group started to dilate at an MAP lower than that of the control group (130 versus 180 mm Hg, respectively). The arterioles constricted when the fistula was occluded at normal MAP. However, when the fistula was occluded at an MAP higher than 130 mm Hg, the vessels dilated. CONCLUSIONS: It was demonstrated that (1) chronic hypoperfusion induced impairment of the upper limit of autoregulation and (2) sudden fistula closure under hypertensive conditions caused vasodilation of the arterioles. These findings suggest that rapid restoration of perfusion pressure is possibly followed by a pressure breakthrough phenomenon in a chronically hypoperfused cerebrovasculature. PMID- 8711808 TI - Cerebroprotective effects of aminoguanidine in a rodent model of stroke. AB - BACKGROUND AND PURPOSE: During a cerebral infarction, a complex cascade of cytotoxic events ultimately determines the volume of brain cell loss. The studies presented here demonstrate that aminoguanidine, an experimental therapeutic currently in clinical trials to prevent diabetic complications, is cerebroprotective in focal cerebral infarction. METHODS: Adult Lewis rats (n = 6 to 12 per group) were anesthetized with ketamine and subjected to focal cerebral infarction by tandem permanent occlusion of the right middle cerebral artery and ipsilateral common carotid artery (CCA), followed by temporary occlusion of the contralateral CCA. Infarct volume (cortical) was assessed 24 hours after the onset of ischemia by planimetric analysis of coronal brain slices stained with tetrazolium. RESULTS: Aminoguanidine (320 mg/kg IP) administered 15 minutes after the onset of ischemia resulted in a significant reduction of infarct volume (7.6 +/- 2.6% of hemisphere in controls versus 1.3 +/- 0.2% of hemisphere in aminoguanidine-treated rats; P < .05). Administration of aminoguanidine conferred significant cerebroprotection even when administered 1 or 2 hours after the onset of ischemia (88% and 85% reduction from control, respectively; P < .05). Cerebroprotection by aminoguanidine was independent of systemic physiological variables known to influence stroke size (eg, temperature, mean arterial blood pressure, blood glucose, and arterial pH, PCO2, and PO2). CONCLUSIONS: These results indicate that the stroke-reducing properties of aminoguanidine are dose and time dependent, with substantial cerebroprotection persisting even with drug delivery up to 2 hours after the onset of ischemia. It is now plausible to pursue development of aminoguanidine as an experimental therapeutic in stroke, and possible mechanisms of these cerebroprotective effects are under consideration. PMID- 8711810 TI - Acute changes in intracranial pressure and pressure-volume index after forebrain ischemia in normoglycemic and hyperglycemic rats. AB - BACKGROUND AND PURPOSE: Hyperglycemia enhances the deleterious effect of global cerebral ischemia. One possible explanation is that increased anaerobic glycolysis leads to exaggeration of intracellular acidosis and increased postischemic edema. To examine the importance of this edema on postischemic cerebral perfusion dynamics, we measured acute changes in intracranial pressure (ICP), specific gravity, and the pressure-volume index (PVI) after forebrain ischemia in normoglycemic and hyperglycemic rats. METHODS: Rats underwent 15 minutes of forebrain ischemia and 90 minutes of reperfusion. ICP and mean arterial pressure were continuously monitored. Before ischemia, rats received either saline or glucose intravenously. Ninety minutes after ischemia, the specific gravity of the neocortex was measured. In a second experiment, the PVI was measured at 20 and 60 minutes after ischemia. RESULTS: Preischemic ICP (mean+/-SD) was 7 +/- 1 mm Hg in both groups. A peak ICP (approximately 11 mm Hg) occurred within 15 to 20 minutes after ischemia in both groups. Between 25 and 80 minutes after ischemia, ICP was significantly but only slightly greater in hyperglycemic than in normoglycemic rats. Cerebral perfusion pressure was similar between groups and remained greater than 100 mm Hg. Specific gravity was also similar for both groups but was less than normal values. The PVI in hyperglycemic rats was lower than in normoglycemic rats, indicating reduced compliance. CONCLUSIONS: These findings indicate that hyperglycemia-augmented intraischemic tissue acidosis does not contribute to worsened outcome by means of compromised cerebral perfusion pressure during the early stages of recovery. Nevertheless, evidence was found for decreased cerebral compliance, indicating an effect of hyperglycemia on intracranial volume compartments other than cortical parenchyma. PMID- 8711811 TI - GABAergic and asymmetrical synapses on somata of GABAergic neurons in CA1 and CA3 regions of rat hippocampus. A quantitative electron microscopic analysis. AB - BACKGROUND AND PURPOSE: CA1 pyramidal neurons in hippocampus die while CA3 neurons survive after transient ischemia. The imbalance of excitation and inhibition may contribute to this selective vulnerability. The purpose of this study was to examine the morphological basis of the above hypothesis. METHODS: Male Wistar rats were perfused with 4% parafor-maldehyde and 0.2% glutaraldehyde in 0.15 mol/L phosphate buffer. Coronal sections (50 microns) cut on a microtome were processed for gamma-aminobutyric acid (GABA) immunocytochemistry. Sections for electron microscopy were postfixed in 0.5% osmium tetroxide and embedded in high-viscosity epoxy resin. Ultrathin sections were cut and observed with an electron microscope. RESULTS: GABA-positive neurons in the stratum pyramidale received more GABAergic synapses than asymmetrical synapses. The percentage of somatic membrane of GABA-positive neurons covered by asymmetrical synapses in the CA1 region (3.17 +/- 1.13%) was higher than that in the CA3 region (2.15 +/- 0.18%, P < .05). The ratio of asymmetrical to GABAergic synapses per 10 microns somatic membrane in the CA1 region (0.71 +/- 0.22) was higher than that in the CA3 region (0.53 +/- 0.14, P < .05). The ratio of the percentage of somatic membrane covered by asymmetrical/ GABAergic synapses in the CA1 region (0.33 +/- 0.14) was also significantly higher than that in the CA3 region (0.20 +/- 0.07, P < .05). CONCLUSIONS: The GABAergic neurons in the CA1 region receive stronger excitatory inputs than those in the CA3 region, which provides a morphological basis for differences in excitability that may contribute to selective vulnerability after transient ischemia. PMID- 8711812 TI - Leukoencephalopathy-related cerebral amyloid angiopathy with cystatin C deposition. AB - BACKGROUND: We have described sporadic cases of cerebral amyloid angiopathy with cerebral hemorrhage showing a low cystatin C level in the cerebrospinal fluid detected by enzyme-linked immunosorbent assay. Recently, several cases of leukoencephalopathy in patients with cerebral amyloid angiopathy have been reported. We describe a sporadic case of leukoencephalopathy with cystatin C-type cerebral amyloid angiopathy diagnosed during life by enzyme-linked immunosorbent assay. CASE DESCRIPTION: A 74-year-old man who had suffered from progressive dementia for 3 years was admitted with right hemiparesis, dysarthria, and ataxia. MRI revealed pontine infarction and multiple lacunar state with leukoaraiosis. We suspected cystatin C-type cerebral amyloid angiopathy because of the low level of cystatin C in the cerebrospinal fluid. The patient died of sepsis 3 months later, and the presence of leukoencephalopathy with cerebral amyloid angiopathy was confirmed by autopsy. Immunohistological examination disclosed cystatin C and beta-protein deposition in amyloid structures of the cortical cerebral arteries. CONCLUSIONS: Measurement of cystatin C in the cerebrospinal fluid by enzyme linked immunosorbent assay is a useful method of diagnosing leukoencephalopathy related to sporadic cystatin C-type cerebral amyloid angiopathy. PMID- 8711813 TI - Adult-onset MELAS. Evidence for involvement of neurons as well as cerebral vasculature in strokelike episodes. AB - BACKGROUND: We report a 46-year-old woman with implications regarding pathogenesis of strokelike episodes in MELAS (mitochondrial encephalomyopathy, lactic acidosis, and strokelike episodes). She had a 10-month history of episodic seizures, strokes, cognitive decline, vomiting, and ileus. She also had sensorineural hearing loss, insulin-dependent diabetes mellitus of several years' duration, and persistent lactic acidosis. Family history was pertinent for a similar syndrome in her deceased mother (onset in her sixties), for hearing loss and diabetes mellitus in two brothers, and for hearing loss in her only child, a son. CASE DESCRIPTION: Serial MRIs of the brain revealed severe but evanescent cerebral cortical abnormalities. A left temporal brain biopsy was performed to exclude encephalitis. Light microscopy revealed a diffuse fibrillary gliosis with abundant reactive gemistocytes, focal evidence of ischemic neuronal injury, and edema. Electron microscopy revealed bizarre enlarged mitochondria and changes consistent with cellular edema. Succinate dehydrogenase staining was strongly reactive within cerebral blood vessels and within neurons. A point mutation was subsequently found at nt 3243 of the mitochondrial tRNA(Leu(UUR) gene in peripheral leukocytes and in brain, confirming the clinical diagnosis of MELAS. Quantitation revealed that 82% of brain mitochondria carried the disease mutation, indicating that most, if not all, tissues were affected. CONCLUSIONS: Our findings suggest that strokelike episodes in MELAS result from defects in neuronal metabolism, as well as in cerebral vasculature. PMID- 8711814 TI - Aneurysm of the left sinus of Valsalva. An unusual source of cerebral embolism. AB - BACKGROUND: Aneurysm of the ascending aorta is rarely reported as the source of emboli. We report a patient with a minor stroke in whom a saccular aneurysm of the left sinus of Valsalva was diagnosed as the presumed source of cerebral embolism. CASE DESCRIPTION: A 49-year-old right-handed woman presented 10 days after sudden onset of right-sided hemiparesis. Her history was uneventful apart from an acute inflammation of the subcutaneous tissue in the right leg 20 years earlier. A diastolic murmur was heard best over the third left intercostal space. Results of duplex ultrasound investigation of the cerebral vessels, chest x-ray, and electrocardiography and biochemical and hematological variables were normal. CT of the brain showed a small hypodense area in the left frontal region. Transthoracic and transesophageal echocardiography detected moderate aortic regurgitation and a saccular aneurysm originating from the left sinus of Valsalva. The aneurysm had calcified walls and contained thrombotic material. Surgical closure of the aneurysm with a pericardial patch was performed to prevent recurrent embolism and rupture. Coaptation of the aortic valves was achieved, and no residual aortic regurgitation could be detected. CONCLUSIONS: We conclude that an unruptured sinus of Valsalva aneurysm should be included in the list of sources of embolism. Transthoracic echocardiography establishes the diagnosis. Transesophageal echocardiography provides additional information about the origin and size of the aneurysm and presence of thrombotic material. Surgical closure of the aneurysm prevents rupture and recurrent embolism and possibly corrects aortic regurgitation. PMID- 8711815 TI - The evolution of surgery for the treatment and prevention of stroke. The Willis Lecture. PMID- 8711816 TI - Concern about safety of carotid angioplasty. PMID- 8711817 TI - Cerebral oximetry in patients undergoing carotid endarterectomy under regional anesthesia. PMID- 8711818 TI - Further debate on the measurement of carotid stenosis. PMID- 8711819 TI - Sensory abnormalities after stroke. PMID- 8711820 TI - Isolated cortical vein thrombosis and activated protein C resistance. PMID- 8711821 TI - [Laparoscopy]. PMID- 8711822 TI - [Current status of laparoscopic surgery in pediatric urology]. AB - The current world-wide experience with laparoscopy in the field of pediatric urology is summarized. Based on significant personal expertise, the operative technique and instrumentation of the different procedures are described. Herein standard indications (i.e., diagnosis and therapy of cryptochidism, bilateral varicoceles) can be distinguished from rather infrequent procedures (i.e., renal cyst resection, nephrectomy) and experimental indications (case reports) such as bladder "auto" augmentation, antirefluxplasty and pyeloplasty. At the referring centers, laparoscopic surgery in pediatric urology has significantly increased. Therefore, this minimally invasive, delicate technique is becoming more popular. PMID- 8711823 TI - [Retroperitoneoscopy. Technique and experiences with the first 100 patients]. AB - Retroperitoneoscopic procedures were already being performed in the late 1970s. The clinical breakthrough of retroperitoneoscopy, however, was initiated in 1992 by the balloon-dissecting technique of Gaur, together with the experience gained with transperitoneal laparoscopic procedures in the upper retroperitoneum. We have modified the balloon-dissecting technique into a hydraulic video-optically controlled balloon dissection of the retroperitoneal space, and this procedure was recently further simplified. From December 1992 to December 1995 we performed 100 retroperitoneoscopic procedures in 98 patients (aged 4-82 years). Twenty-two patients had undergone previous abdominal surgery, 16 patients had been operated on for kidney and ureter problems. We performed 28 simple procedures (6 renal biopsies, 17 renal cyst resections, 4 ureterocutaneostomies, 1 foreign body retrieval), 65 difficult operations (47 nephrectomies, 5 nephroureterectomies, 5 nephropexies, 4 ureterolithotomies, 2x ureterolysis). There were also 9 complicated cases (5 heminephrectomies, 2 tumor nephrectomies, 2 dismembered pyeloplasties). Operating time, complications and conversion rate to open surgery mainly depended on the difficulty of the procedure and the personal learning curve, resulting in 50-90 min, 0% and 3.8%, respectively, for an easy retroperitoneoscopy, 90-210 min, 12% and 9.2% respectively for a difficult operation, and 180-390 min, 22% and 11%, respectively, for a very difficult procedure. Excluding the 17 patients with complications or conversion (unclear anatomy, extremely difficult dissection), the mean postoperative need for analgesics was 0.7 vials, and the mean hospital stay was 4.7 days. We have now passed most of our learning curve, and retroperitoneoscopy has become a standardized procedure that is a part of the training program for the experienced urologist. PMID- 8711824 TI - [Laparoscopic spermatic vein ligation]. AB - Between December 1991 and November 1995 a total of 231 patients underwent laparoscopic varix ligation in our department. The laparoscopic varicocelectomy offers a lower rate of morbidity, allows for microscopic dissection with preservation of the spermatic artery and is amenable to bilateral ligation without a second incision. The spermatic arteries were identified by using a laparoscopic vascular Doppler probe. Any laparoscopic procedure carries some risk of injury to intra-abdominal organs when placing the primary trocar. Therefore, a new extraperitoneal approach has been developed to avoid these risks. PMID- 8711825 TI - [Laparoscopic and retroperitoneoscopic kidney pyeloplasty]. AB - The aim of this study was to evaluate laparoscopic and retroperitoneoscopic pyeloplasty and to compare the efficacy of dismembered and non-dismembered techniques. Between April 1993 and December 1995 a modified laparoscopic transperitoneal (18 patients) and a retroperitoneoscopic approach (3 patients) were used for the management of ureteropelvic junction obstruction. In 11 patients aberrant vessels were encountered; one patient had a horse-shoe kidney. Surgical repair was achieved by dismembered pyeloplasty (8 patients), non dismembered Fenger-plasty (longitudinal incision-transverse closure: 7 patients), transection and reanastomosis of the renal pelvis (1 patient), ureterolysis and displacement of ventrally crossing vessels (4 patients). In one patient dismembered pyeloplasty could not be scheduled because of cardiovascular problems. A minimal transient lesion of the sympathetic nerve was observed postoperatively in one patient and pulmonary embolism in another. The operative time in dismembered pyeloplasty was between 240 and 360 min (mean 280); the results were good in all patients. Equally good results were obtained with non dismembered Fenger-plasty, and the operating time was shorter (120-180 min). Ureterolysis was found to have a failure rate of 50%. Laparoscopic dismembered pyeloplasty yielded good results, but it is too complicated to become a standard procedure. Non-dismembered Fenger-plasty, which also showed good results, is more suitable for laparoscopy and retroperitoneoscopy. The indications for this technique should be defined more precisely as more experience is collected. The results of ureterolysis-when used as a single measure-were poor, and therefore this technique should be abandoned. PMID- 8711826 TI - [Laparoscopic technique--which developments are possible?]. AB - The progress of laparoscopy is influenced by both the medical and technical aspects. The development of endoscopes and various rigid instruments has increased the indication options. Nevertheless, several drawbacks remain, e.g. the limited spatial view, the missing sense of touch, and reduced mobility in the operation area. New 3D visual systems now introduce spatial view. Flexible instruments are being developed that allow thorough examination of organs. While these enhancements are now becoming available, research and development are making progress and preparing the next steps. One vision is the development of a telepresence and telemanipulation system. With it, at the patient's side we will have an endoscope guidance system and several instrument guidance systems, which will be telemanipulated from a control station. At the control station, a 3D picture from the operation scene, together with virtual reality simulation pictures will be available. Force reflection as well as palpatory sensing information will be readily available to the telesurgeon. These new developments will improve the quality of the surgery for the benefit of both the patient and surgeon. Furthermore, the training of new surgeons will be eased by the use of sophisticated simulators using virtual reality techniques. These and further technical developments will not only lead to an improvement in current laparoscopy procedures, but it can be expected that additional procedures will be developed that are not yet possible and accessible to laparoscopy. PMID- 8711827 TI - [Transperitoneal and retroperitoneal laparoscopic nephrectomy in comparison with conventional nephrectomy]. AB - We report on our clinical experience with laparoscopic nephrectomy in 35 patients. We have performed a total of 18 transperitoneal laparoscopic nephrectomies (TLN) for benign renal disease. After the introduction of a hydraulic dissecting device, we have since performed 17 retroperitoneal laparoscopic nephrectomies (RLN). These data were compared with those in 19 consecutive open nephrectomies (N). All groups were comparable in terms of indication. The mean operative time for benign disease was 206.5 min for TLN, 211.2 min for RLN and 117 min for N. Analgesic medication requirement per patient was 2 days for TLN, 1 day for RLN and 4 days for N, while the postoperative hospital stay averaged 7 days for TLN, 6 days for RLN and 10 days for N. Our results demonstrate the advantage of a laparoscopic approach over open surgery and also reveal the distinct benefit of a retroperitoneal approach. However, due to the small number of indications this procedure should be restricted to a few urologic centers. PMID- 8711828 TI - [Laparoscopic operations in urologic clinics--results of a survey]. AB - This paper summarizes the results of a questionnaire initiated in August 1995 by the Arbeitskreis "Minimal invasive Urologie" of the Deutsche Gesellschaft fur Urologie. We received 235 answers from a total of 380 departments, which were queried by fax. Laparoscopic surgery is currently performed in 36% of all answering departments and in 75% of all university divisions. However, only 5% of the departments do more than 30 procedures per year. The most frequent indications are cryptorchisdim in 24%, pelvic lymph node dissection in 20%, lymphoceles in 16%, renal cyst resection in 15%, varicocelectomy in 14%, and laparoscopic nephrectomy in 11% (30% of all laparoscopically active units). Since laparoscopy in urology does not have an indication for a high number of patients (and departments), knowledge about how to use this minimally invasive technique will be further concentrated in already existing centers, as this guarantees sufficient frequency of its use and expertise. PMID- 8711829 TI - [The future of laparoscopic surgery in urology]. AB - Laparoscopy has followed the typical phases in the development of any new operative technique. We have now begun to realize its limitation and are beginning to standardize the procedures and specify their indications. Since laparoscopy predominantly reduces the trauma of access, the development of a total operative trauma score has assisted us in determining the impact of specific laparoscopic procedures compared to their open alternatives. Laparoscopy is responsible for less blood loss, less postoperative pain, a shorter hospital stay and a very small scar, all of which accounts for its popularity among patients. A rational combination of laparoscopy, endoscopy and open surgery to treat urological diseases should be developed in the future. PMID- 8711830 TI - [Laparoscopic adrenalectomy. Experiences with 50 patients]. AB - Fifty patients with adrenal tumors (18 men and 32 women, average age 51.8 years) underwent laparoscopic adrenalectomy from February 1992 to October 1995. Clinical diagnosis included pheochromocytoma in 3 patients, primary aldosteronism (including 1 with a 11-OH-corticosterone-producing tumor) in 15, Cushing's syndrome (including 7 with pre-Cushing's syndrome) in 13, non-functioning tumors in 17, 1 metastatic adrenal carcinoma, and 1 adrenal tuberculosis. Transperitoneal laparoscopic adrenalectomy was performed by the method reported previously. Extraperitoneal laparoscopic adrenalectomy was performed with the patient under general anesthesia in the lateral position. A working space was created by inserting a balloon dissector through a small skin incision. A total of four trocars were inserted. Three of the 50 patients were switched to open surgery, including 1 with metastatic adrenal carcinoma and 1 with adrenal tuberculosis. Laparoscopic tumor removal was successful in the other 47 patients. The average operating time and blood loss were 209 min and 177 ml, respectively. Blood loss was greater in the patients with pheochromocytoma. In patients with Cushing's syndrome, postoperative recovery tended to take longer. Postoperative complications occurred in 40% of the patients in this group, but all complications were minor and successfully treated without any surgical procedures. In the 10 patients undergoing retroperitoneal laparoscopic adrenalectomy, operative courses were excellent, excluding 1 patient with adrenal tuberculosis. Although laparoscopic adrenalectomy is considered to be appropriate for patients with pheochromocytoma and Cushing's syndrome, it appears unsuitable for the removal of malignant and inflammatory lesions. PMID- 8711831 TI - [Complications of laparoscopic interventions in urology]. AB - From April 1987 to November 1995, we performed 741 laparoscopic procedures, in which we observed 14 major complications (1.9% complication rate). The complication rate was 4% in the first 100 procedures--more than twice as many as in the following 631 procedures, which had a complication rate of 1.6%. Although between 1987 and 1990 laparoscopy was mainly used for the diagnosis of cryptorchidism, we then progressed to therapeutic measures with the help of the laparoscopic technique. After performing laparoscopic orchiectomy (1990), we introduced fenestration of lymphoceles (1991), varicocelectomy (1991), pelvic staging lymphadenectomy (1992), nephrectomy (1992), and adrenalectomy (1994) into clinical practice, in addition to other, more rarely occurring procedures. The complications we observed demonstrate a clear learning curve and show that critical, gradual practice of the laparoscopic technique is necessary. PMID- 8711832 TI - ["Contra laparoscopy" position in treatment of testicular varicocele]. AB - Laparoscopic varicocelectomy does not offer any significant advantage in comparison to sclerotherapy or open surgical procedures. Invasiveness (transperitoneal approach, intraoperative complications, general anesthesia, duration of operation, possible ligation of the spermatic artery) is not less, but the technical expenditure and costs are higher than those in competing procedures. Laparoscopic treatment is not the procedure of first choice and even in recurrent varicocele it should only be performed if prior treatment by repeated antegrade sclerotherapy is not successful and distal reflux has been ruled out. Therefore, the laparoscopic approach is of only minor value in the treatment of varicocele testis. PMID- 8711833 TI - [Urothelial carcinoma of the navicular fossa]. AB - We report a case of transitional cell carcinoma arising in the fossa navicularis. The patient was a 74-year-old man. He had no history of sexually transmitted disease or urethral stricture. Clinically, the tumor was suspected to be a condyloma acuminatum; however, the pathological diagnosis yielded an unexpected result: transitional cell carcinoma. Radiological examinations and cystoscopy showed no other tumor besides the primary cancer in the fossa navicularis. Partial resection of the urethra was performed and the patient has been without evidence of disease for 3 years. PMID- 8711834 TI - [Traditional and future criteria for progression in renal cell carcinoma. Molecular biology and clinical aspects]. AB - In addition to the commonly used pathologic staging systems such as the TNM classification, the grading and demographic features have been reported to affect survival following surgical extirpation of renal cell carcinoma (RCC). These features, especially the pathologic stage, are well established as prognostic factors. However, several cellular and molecular variables, although potentially important in the ultimate outcome, are not taken into consideration by these criteria. Thus patients with different prognoses may be classified as belonging to the same stages. Attempts are now made to use cytogenetic and molecular findings to predict long-term survival of RCC patients. Chromosome 16 q and 14 q aberrations may play an important role in the future by identifying the high-risk groups of patients with papillary and nonpapillary RCC, resp. In nonpapillary RCC, mutations of the von Hippel-Lindau gene have been implicated as the initial step of carcinogenesis. However, the subsequent steps remain to be elucidated and the search for genetic markers associated with tumor progression is under way. The distinction between patients with high and low risk of progression will become increasingly important as more effective adjuvant therapies are available. PMID- 8711835 TI - [Hyperuricemia]. PMID- 8711836 TI - [Use of YAG-ND laser in the treatment of not healing trepanation cavities after cleansing surgery of the middle ear]. AB - Not healing trepanation cavities after middle ear surgery were exposed to YAG-Nd laser radiation (1.06 and 1.32 mk). Persistent epidermization and discontinuation of the discharge were achieved in 72% of the patients with postoperative otorrhea. Optimal regimens of laser radiation (wave length, exposition, power) are detailed for various forms of the pathological process. PMID- 8711837 TI - [Features of the status of otorhinolaryngologic organs of subway train mechanics]. AB - 337 mechanics exposed to chemical and physical hazards while servicing subway trains were examined for ENT status. Abnormalities were registered in 87.5% of the examinees. Frequent isolated and combined affections of the nose and pharynx occurred as a rule with mucosal dystrophy. Allergic conditions were not evident. 58 patients with hearing defects had cochlear neuritis of occupational origin. Prophylactic measures are proposed: individual aids protecting hearing organ and respiratory tracts, improved routine medical examinations with indication of the risk groups. PMID- 8711838 TI - [Features of changes in otorhinolaryngologic organs of operators of automatic plasma tube cutting]. PMID- 8711840 TI - [Several aspects of treatment and prevention of chronic laryngeal stenosis]. AB - The occurrence of scar laryngostenosis due to traumas and removal of laryngeal tumors is now on the increase. This negative trend may be corrected by rejection of long-term intubation, immediate conversion of crico-, conico- and thyrotomy into tracheostomy, employment of preventive laryngostomy in initial treatment of patients with serious trauma of the larynx. It should be taken into consideration that in restoration of respiration through natural airways bronchopulmonary complications, dysphonia, dysphagia, hair growth into the laryngeal lumen are possible. PMID- 8711839 TI - [Differential diagnosis of unilateral paresis and paralysis of the larynx]. AB - Differential diagnosis of laryngeal paresis and paralysis using electron glottography (EGG) was tried in 57 patients and 30 healthy controls. Measurements were made of speed quotient (SQ) and open quotient (OQ) at piano and forte. It was found that in patients and controls SQ and OQ were not similar. SQ and OQ differed also in paralysis and paresis. Tonicity of the inner laryngeal muscles seems to decline more in paralysis than in paresis. Aerodynamic properties of the larynx are damaged more in paralysis and remain unchanged in paresis. Reduced SQ and high OQ at forte indicate unilateral laryngeal paralysis. OQ should be measured both at piano and forte. Thus, EGG is an effective procedure for differential diagnosis of laryngeal paralysis and paresis. It is especially convenient when laryngostroboscopy is problematic. PMID- 8711841 TI - [Blast transformation activity of autologous saliva in patients with different clinical forms of chronic bronchitis]. AB - The study of mitogenic effect of autologous saliva in respect to peripheral blood lymphocytes of patients with different clinical forms of chronic tonsillitis (CT) discovered that saliva from all the examinees was highly active as regards blast cell transformation. The intensity of lymphocyte blast-transformation in the presence of autologous saliva grows with CT transformation from compensated to decompensated form and insignificantly declines in clinical symptoms of autoimmune disease. The saliva mitogenic effect is probably stimulated by the whole complex of biologically active substances which by provoking hypertrophy of the lymphoid tissue contributes to autoimmunization of the body. It is proposed to use lymphocyte blast-transformation with autologous saliva for diagnosis of subclinical autoimmune diseases. PMID- 8711842 TI - [Effects of pathology of the gastrointestinal system on the state of the mucous membrane of the larynx and pharynx]. AB - The hypophysis on contribution of gastrointestinal pathology to development of chronic pharyngo-laryngitis (CPL) was investigated in the study of 110 patients with catarrhal, hypertrophic, atrophic and mixed CPL (20, 12.7, 27.3, 40% of patients, respectively). Gastroenterological examination revealed gastritis plus gastroduodenitis, gastroduodenal ulcer, biliary and pancreatic disease, combined gastroduodenal pathology in 21.8, 30.9, 20, 27.3% of patients, respectively. Combined therapy including treatment of gastrointestinal disorders, produced stable remission of pharyngo-laryngitis in 97.3% of the patients treated. Thus, it is evident that a close relationship exists between pharyngolaryngeal and gastrointestinal affections. PMID- 8711843 TI - [Endoscopic diagnosis of multiple primary cancer of the upper airways]. AB - The authors analyze the results obtained at comprehensive examination of 123 patients with multifocal synchronous and metachronous cancer of the upper respiratory tracts and the lungs. Cancer of the larynx and the lungs was registered in 107 patients, multifocal cancer of the pharynx and lungs in 16 patients. Synchronous and metachronous cancer of the larynx and the lungs was diagnosed in 48 (44.9%) and 59 (55.1%) patients, respectively. High efficacy of endoscopic diagnosis is shown for multifocal cancer of the larynx and lungs, pharynx and lungs. PMID- 8711844 TI - [Abscess tonsillectomy in acute parapharyngitis]. PMID- 8711845 TI - [Effectiveness of bioparox treatment in acute inflammatory diseases of the upper airways]. AB - Acute inflammation of the upper respiratory tract was treated with bioparox in 40 patients and conventionally in 20 patients. The assessment of the treatment effect was carried out by subjective and objective responses, rhinomanometry, trends in cell composition and phagocytic activity of leukocytes in smears from nasal mucosa. The data obtained provided evidence for high efficacy of bioparox which promoted faster regress of subjective and objective manifestations of the inflammation. PMID- 8711846 TI - [Use of rovamycin and amox-clav in patients with infectious-inflammatory pathology of the ORL organs]. AB - Clinical and microbiological findings led the authors to the conclusion that antibacterial drugs rovamycin and amox-clav are active against ENT infection and inflammation. The trial included 69 and 42 patients given rovamycin and amox clav, respectively. PMID- 8711847 TI - [Use of benzofurocaine in stimulation of regenerative processes in the surgical treatment of patients with chronic suppurative otitis media]. AB - Healing of postoperative wounds of the middle ear treated with benzofurocaine was analysed in 57 patients. The drug was given for 10 postoperative days as 10 ml intramuscular injections of 1% solution twice a day. Benzofurocaine-treated patients exhibited less extensive edema of the parotid tissues and walls of the external acoustic meatus, faster epidermisation of the trepanation cavity and retention of the neotympanum graft. Cytochemically, intramuscular injections of benzofurocaine enhance functional activity of epitheliocyte nuclei as shown by reduced DNA anisotropy and intensified cell utilization of glycogen and release of cationic protein. PMID- 8711848 TI - [A stab wound of the neck with injury of the common carotid artery]. PMID- 8711849 TI - [Case report of foreign bodies of the esophagus]. PMID- 8711850 TI - [Low-intensity laser irradiation in otorhinolaryngology]. AB - Low-intensity laser irradiation is a modality widely practiced in otorhinolaryngology. Basing on extensive personal experience and literature data the authors provide ENT specialists with relevant techniques and parameters of the radiation for different otorhinolaryngological diseases. PMID- 8711851 TI - [Synchronous-metachronous malignant neoformations of the larynx and kidneys]. PMID- 8711852 TI - [Nasal carbuncle complicated by cavernous sinus thrombosis with fatal outcome]. PMID- 8711853 TI - [A case of malignant mixed tumor of the larynx]. PMID- 8711854 TI - [Large osteoma of the frontal sinus]. PMID- 8711855 TI - Feline lower urinary tract disorders. Definition of terms and concepts. AB - Lower urinary tract disorders of male and female cats may be caused by a variety of fundamentally different causes. The term Feline urologic syndrome should be abandoned and substituted with descriptive etiopathogenic terms whenever possible. If the underlying cause cannot be identified, the term Idiopathic lower urinary tract disease is recommended. PMID- 8711856 TI - Applied anatomy and physiology of the feline lower urinary tract. AB - Paired ureters, urinary bladder, and urethra constitute the lower urinary tract. Oblique passage of ureters through the bladder wall results in compression of the distal ureter to preclude urine reflux. Ureters are anchored by longitudinal ureteral musculature that outlines the bladder trigone and extends into dorsal submucosa of the urethra as urethral crest. The urinary bladder can be divided into apex, body, and neck. The male urethra has penile and pelvic components, the latter is divisible into preprostatic, prostatic, and postprostatic regions. The muscle coat of the bladder-urethra forms three functional entities in craniocaudal series. These are the detrusor muscle (to effect voiding), internal urethral sphincter (smooth muscle for generating tonic resistance), and external urethral sphincter (striated urethralis m. for phasic and voluntary continence). The vesical neck is a transition region. It is part of the internal urethral sphincter by virtue of its histology and innervation, but it contains detrusor fascicles that pull it open during micturition. Viscous accommodation plus sympathetic reflex inhibition of the vesical wall allows the urinary bladder to greatly expand in volume with minimal increase of intravesical pressure, within limits. At low volumes continence can be maintained by passive resistive elements of the urethral outlet. As volume increases, sympathetic reflex activity is necessary for continence. The striated external urethral sphincter is reflexly contracted to counter abrupt elevations of intravesical pressure and to maintain continence voluntarily. The pelvic plexus conveys sympathetic and parasympathetic innervation to the urinary tract. The pudendal nerve supplies the urethra and urethralis muscle. Ureters are largely independent of innervation. Internal and external urethral sphincters are activated by spinal reflexes, sympathetic and somatic reflexes, respectively. Normal micturition (sustained detrusor contraction and sphincter inhibition) is a brainstem-driven reflex, involving a spino-bulbo-spinal pathway and a pontine micturition center that switches from urine storage to micturition. All of the reflexes depend on neural activity in tension mechanoreceptors of the bladder wall and sacral afferent fibers. PMID- 8711857 TI - Healing responses of the lower urinary tract. AB - Healing of the urinary tract follows a series of events similar to other epithelial surfaces. This process, however, occurs more rapidly in the urinary tract. A variety of factors that can affect the healing response of the urinary tract are presented in this article. It is important for the veterinarian to be aware of these factors so that the appropriate surgical or medical treatment can be provided to the patient to optimize the healing processes. PMID- 8711858 TI - Biologic behavior of feline lower urinary tract diseases. AB - This case represents the typical presentation and natural progression of idiopathic lower urinary tract disease in cats. As depicted, it generally occurs in young cats. Cats often have evidence of hematuria with minimal or no leukocyturia. In this cat, clinical signs and hematuria subsided in 3 days. Tristan was evaluated monthly for 1 year. During this period, a recurrence of clinical signs or hematuria did not occur. PMID- 8711859 TI - Feline urolithiasis. Etiology and pathophysiology. AB - A variety of different types of uroliths occur in cats, of which calcium oxalate and magnesium ammonium phosphate are the most common. Treatment and prevention are most likely to be successful if associated risk factors are identified and eliminated or controlled. PMID- 8711860 TI - Feline urethral plugs. Etiology and pathophysiology. AB - Feline urethral plugs commonly are composed of large quantities of matrix mixed with minerals (especially struvite). However, some urethral plugs are composed primarily of matrix, some consist of sloughed tissue, blood, and/or inflammatory reactants, and a few are composed primarily of aggregates of crystalline minerals. The formation of matrix-crystalline urethral plugs may be analogous to the preparation of fruit jello. PMID- 8711861 TI - Pathophysiology of urethral obstruction. AB - Obstructive uropathy refers to abnormalities in structure or function of the urinary tract caused by impairment of normal flow of urine, and the resulting local and systemic effects of that impairment. Clinic consequences of obstructive uropathy are associated with abnormalities in fluid balance, electrolyte metabolism, acid-base balance, and retention of metabolic wastes. These consequences are partly due to build up of intravesical, ureteral, and renal pressure, influx of leukocytes into renal parenchyma resulting in release of cytokines, and alterations in intravascular hemodynamics. This article discusses pathophysiologic mechanisms and consequences of obstructive uropathy in cats. PMID- 8711862 TI - Inherited and congenital diseases of the feline lower urinary tract. AB - Congenital urinary tract disorders of young cats may result from heritable (genetic) or acquired disease processes that affect differentiation and growth of the developing urinary tract, or from similar disease processes that eventually affect the structure or function of the mature urinary system. Although congenital diseases of the feline lower urinary tract are uncommon, clinical signs associated with these anomalies may be indistinguishable from those of other acquired causes of lower urinary tract disease. Early detection and proper management of congenital disorders may result in restoration of urinary bladder and urethral function and/or progressive urinary tract dysfunction. PMID- 8711864 TI - Bacterial urinary tract infections. AB - Bacterial urinary tract infection rarely is the initial cause of lower urinary tract disease in cats; however, a substantial number of cats with various lower urinary tract disorders develop urinary infections as a complication of the underlying disease or its treatment. Examination of urine sediment for pyuria is a good screening test for urinary tract infection, but results of urine cultures are needed to diagnose such infections reliably, as well as to guide therapeutic decisions. Appropriate antimicrobial drug therapy is the cornerstone of treatment for urinary tract infection. Treatment of the underlying urinary disorder also is important, and follow-up urine cultures should be performed to determine therapeutic outcome. PMID- 8711863 TI - Viral infections of the feline urinary tract. AB - The exact cause of hematuria, dysuria, and urethral obstruction remains unknown in a large percentage of naturally occurring cases of feline lower urinary tract disease (FLUTD). One attractive hypothesis implicates viruses as the cause of some idiopathic forms of FLUTD; supporting this hypothesis is the fact that a gamma herpesvirus, a calicivirus, and a retrovirus have been isolated from urine and tissues obtained from cats with this type of disease. Although the clinical course and laboratory findings of cats with idiopathic FLUTD are suggestive of an infectious cause, the question of whether viruses have a pathologic role in some forms of naturally acquired FLUTDs has not been completely answered. PMID- 8711865 TI - The role of Mycoplasma species and Ureaplasma species in feline lower urinary tract disease. AB - The cause of feline lower urinary tract disease (FLUTD) is frequently difficult to determine and may be multifactorial. A controlled cohort study was performed to examine the role of Mycoplasma species and Ureaplasma species in FLUTD. Based on culture and serologic criteria, these microorganisms did not appear to be involved in the group of cats studied. At this time, there is no evidence to implicate Mycoplasma species and Ureaplasma species in FLUTD. PMID- 8711866 TI - Fungal infections of the feline lower urinary tract. AB - Many forms of therapy currently used for empiric treatment of feline lower urinary tract diseases represent significant predisposing factors for fungal infection (i.e., prolonged antibiotic and/or glucocorticoid therapy, aciduria, and indwelling transurethral catheters). Therapy of fungal urinary tract infection should be based on the cat's clinical history, physical examination, and laboratory findings. Amelioration of risk factors has been associated with spontaneous resolution of funguria. For other patients, alkalinization and administration of antifungal chemotherapy are needed to control disease. PMID- 8711867 TI - Interstitial cystitis in cats. AB - Despite much research on FUS, there still is no consensus regarding its cause, or even its definition. We recently have demonstrated that some cases of FUS are similar to interstitial cystitis in human beings. Exclusion of anatomic defects, behavior abnormalities, neoplasia, urolithiasis, urethral obstruction, and urinary tract infection leads one to the diagnosis of interstitial cystitis in cats. This diagnosis suggests the need for new approaches to treatment, including pain management and environmental modification. PMID- 8711868 TI - Detrusor-sphincter dyssynergia. AB - Detrusor-sphincter dyssynergia refers to failure of the urethral sphincter to relax during detrusor contraction. The cause is a central nervous system lesion located between the brain stem micturition center and the sacral spinal cord. This is an extremely rare condition in cats. It may be confused with a failure of urethral relaxation due to local urethral causes such as inflammation or edema. This article reviews detrusor-sphincter dyssynergia to allow the reader to distinguish this rare condition from more common conditions that prevent bladder emptying. PMID- 8711869 TI - Overview of diagnosis of feline lower urinary tract disorders. AB - Disorders of the lower urinary tract are common in cats. Optimal patient care depends on an accurate diagnostic assessment of clinical information and thoughtful analysis of the trade offs between the benefits and risks of further testing and treatment. Systematic approaches to data collection can minimize diagnostic uncertainty and maximize diagnostic efficiency. PMID- 8711870 TI - Cystocentesis. Diagnostic and therapeutic considerations. AB - The diagnostic and therapeutic value of cystocentesis has been recognized for over 80 years. In feline patients with nonobtrusive lower urinary tract diseases (LUTD), normal patients, or patients with nonurinary disorders, diagnostic cystocentesis circumvents many of the potential problems associated with collection of urine specimens by normal micturition, manual compression of the urinary bladder, or catheterization. In patients with obstructive LUTDs, therapeutic cystocentesis temporarily halts the adverse effects of obstructive uropathy and may provide additional time to remove or bypass the obstructive lesion. In our experience, cystocentesis has been associated with few significant side effects; however, it may induce mild transient microscopic hematuria which may be indistinguishable from pathologic hematuria associated with many naturally occurring feline LUTDs. PMID- 8711871 TI - Measurement of the influence of diet on feline urinary pH. AB - The kidneys eliminate the acid produced each day by metabolism and diet. Consequently, urinary pH is influenced by diet. The urinary pH of cats consuming a natural habitual diet consisting of small mammal carcasses is between 6.2 and 6.4. Although the only authoritative way to determine the effect of diet on urinary pH is a feeding trial, there is no universally accepted protocol for measuring urinary pH. When interpreting the urinary pH of a spot or random sample, it is necessary to consider when the sample was obtained relative to time of eating. PMID- 8711872 TI - Feline crystalluria. Detection and interpretation. AB - Crystalluria results from oversaturation of urine with crystallogenic substances. However, oversaturation may occur as a result of in vivo and in vitro events. Therefore, care must be used not to overinterpret the significance of crystalluria. Evaluation of urine crystals may aid in (1) detection of disorders predisposing cats to urolith or matrix-crystalline urethral plug formation; (2) estimation of the mineral composition of uroliths or urethral plugs; and (3) evaluation of the effectiveness of medical protocols initiated to dissolve or prevent urolithiasis. PMID- 8711873 TI - Urolith analysis. Submission, methods, and interpretation. AB - The detection, treatment, and prevention of the causes underlying urolithiasis depend on knowledge of the composition and structure of the entire stone. Therefore, proper and complete analysis of uroliths is an important part of proper patient care. This article discusses methods of urolith retrieval, proper sample submission, methods of urolith analysis, and interpretation of results. PMID- 8711874 TI - Diagnostic imaging of the feline lower urinary tract. AB - Survey radiography, double contrast cystography, and antegrade urethrocystography are valuable imaging tools available to the clinician for evaluation of cats with lower urinary tract disease. However, to be cost effective during patient evaluation, diagnostic radiographic procedures should be used only when needed. An algorithm is presented that describes when radiographic imaging techniques may be most cost effective in providing useful information during the clinical workup of cats with lower urinary tract disease. PMID- 8711875 TI - [Occurrence of chlorinated pesticides and polychlorinated biphenyls in the stalls and outdoor environment in pig-fattening farms]. AB - Contents of some pesticides (lindane, DDT sum, o,p'-DDE, p,p'-DDE, o,p'-DDD, p,p' DDD, o,p'-DDT, p,p'-DDT) and polychlorinated biphenyls (Delor 103, Delor 106, PCB congeners: 28, 52, 101, 118, 138, 153, 180) were investigated on three farms of fattened pigs (D., M. and T) in Hodonin district in 1994, as occurring in feed mixtures (n= 15), drinking water (n = 10), stable sedimented dust (n = 14), slurry (n = 10) and pavement dust (n = 5). Maximum permissible contents (MPC) of extraneous matters in feed mixtures were exceeded in lindane (two samples) and polychlorinated biphenyls (PCB). MPC of extraneous matters were not exceeded in drinking waters in any of the samples examined. Feed mixtures were found to have the highest average lindane content (0.054 mg/kg) and PCB sum (0.24 mg/kg); the ratio of lower chlorinated biphenyls (Delor 103) to higher chlorinated biphenyls (Delor 106) was 12:1; out of the PCB congeners, the highest amounts were determined for congeners 28 (0.067 mg/kg) and 52 (0.022 mg/kg); among the DDT isomers, isomer p,p'-DDT was prevailing (0.008 mg/kg). Drinking water had the 1:1 ratio of lower chlorinated to higher chlorinated biphenyls; out of the PCB congeners, the highest amounts were found out for congeners 28 (2.574 ng/l) and 101 (2.066 ng/l); among the DDT isomers, isomer p,p'-DDE was prevailing (1.887 ng/l). Stable dust was found to have the highest average level of DDT sum (0.026 mg/kg); the ratio of lower chlorinated to higher chlorinated biphenyls was 1:7; out of the PCB congeners, the highest amounts were determined for congeners 138 (0.036 mg/kg) and 153 (0.030 mg/kg); among the DDT isomers, isomer p,p'-DDE was prevailing (0.012 mg/kg). Slurry showed the 1:6 ratio of lower chlorinated to higher chlorinated biphenyls; out of the PCB congeners, the highest amounts were found out for congeners 138 (0.016 mg/kg dry matter) and 153 (0.014 mg/kg dry matter); among the DDT isomers, isomer p,p'-DDT was prevailing (0.005 mg/kg dry matter). Pavement dust had the 1:5 ratio of lower chlorinated to higher chlorinated biphenyls; out of the PCB congeners, the highest amounts were found out for congeners 138 (0.016 mg/kg) and 153 (0.012 mg/kg); among the DDT isomers, isomer p,p'-DDE was prevailing (0.005 mg/kg). Examination of feed mixtures and drinking waters will provide basic data on pollutant inputs to the stable environment. Stable sedimented dust is considered as an outstanding indicator of pollutant contamination of stable environment, regardless whether it comes from feed or other sources. Slurry examination will provide additional information on the total pollutant load of pig farms, informing us about outputs of extraneous matters residues from pig farms. Pavement dust examination indicates the level of pollutant load of outside stable environment as the pollutants can occasionally penetrate to the internal stable environment. PMID- 8711876 TI - [Contamination in areas of Slovakia with strains of Coxiella burnetii imported from abroad]. AB - Following the upsurge of Q fever epidemic in the village Jedl'ove Kostol'any, Nitra district, Slovakia, in April 1993, the possibility of the penetration of coxiellae into free nature was investigated for the next two years. Micromammals and their ectoparasites, primarily ticks, were employed as indicators of the presence of the infection in nature. The infestation of mammals and their ectoparasites was followed in J. Kostol'any on the farm where the goats were housed as well as in its close proximity. The infestation of ticks with coxiellae was further followed in four localities of the Rimavska Sobota district, where the goats after the eradication of Q fever outbreak in J. Kostol'any were transferred for breeding. The presence of coxiellae was not proved in these investigated localities. It is suggested that the pathogenicity of imported C. burnetii strains through Bulgarian goats was decreased under local environmental conditions including the ability to circulate freely in nature or these strains found inconvenient conditions for maintaining their viability. PMID- 8711877 TI - [Clinical and metabolic response in goats with iodopenia after administration of potassium iodide]. AB - A clinical picture and dynamics of metabolic indicators were examined in 40 goats suffering from iodopenia; the examinations took place in the course of potassium iodide application. Crystalline kalium iodatum was administered to all animals in barley groats at a dose of 0.5 g per animal/day within three weeks; then a three week withdrawal followed and the treatment was repeated within the same time period. The thyroid gland consistency started to change at the end of the 3rd week of treatment when the tough elastic to tough thyroid became the soft one. The goiter started to disappear gradually from the 8th week of experiment and the physiological state of this organ was normal in most animals in the 11th week. Among the biochemical indicators, the most sensitive response to treatment was recorded in iodine and thyroxine concentrations in blood serum since their significant increase in comparison with the values before potassium iodide application was obvious on day 14 of treatment (P < 0.01). Return of total lipids and cholesterol content to the physiological limits occurred within two weeks while in glucose it was in the 7th week of treatment. A significant increase in serum immunoglobulins was recorded from day 14 to the end of experiment (P < 0.01). Among the minerals analyzed, positive responses in blood serum to potassium iodide intake were determined in the concentrations of calcium, phosphorus, copper and zinc. Application of potassium iodide to affected goats had positive effects on the dynamics of leukocytes from the end of the 2nd week (P < 0.01), of erythrocytes from the 7th week (P < 0.01), of hemoglobin and hematocrit value at the end of experiment (P < 0.05 and P < 0.01, resp.). PMID- 8711878 TI - [Complications during myelographic examination in dogs]. AB - The occurrence of spinal disease in dogs is quite frequent. Clinical examination and survey radiography do not allow to make an exact diagnosis in many cases. Therefore the radiographic method-myelograhy is used to make an exact diagnosis and to locate the pathological process in the spinal cord. Myelography can be accompanied by some complications caused by a contrast medium of anaesthesia. In this study, the occurrence of complications was evaluated when the contrast medium Dimer X and three types of general anaesthesia were used: xylazine ketamine, chlorpromazine-piritramide, chlorpromazine-pentobarbital. Certain relations between the origin of complications, dog weight and total time of anaesthesia are indicated. The results obtained show that the highest number of cases with complicated regeneration was observed after the use of the combination xylazine-ketamine when the average duration of anesthesia was 31.1 minutes comparing to the combination chlorpromazine-pentobarbitale with average duration of anaesthesia was 127.7 minutes with the lowest number of complications. We also refer to certain relations between weight and the occurrence of complications where the highest correlation (r = 0.59) was recorded for xylazine-ketamine anaesthesia. The correlation coefficient (r = 0.27) for chlorpromazine pentobarbital documents very low correlations of the followed values. PMID- 8711879 TI - A roebuck skull anomaly--a case report. AB - The described roebuck was primarily affected by a prenatal defect in the region of the frontal splanchnocranium. The defect negatively affected the function of the jaws; marked changes in the skull developed secondarily. Other changes in the whole organism resulted from the impaired capability of receiving and processing food. PMID- 8711880 TI - Electron microscopy--tool to diagnostic pathology. AB - This review suggests electron microscopy applications, particularly for leucocyte surface antigens, tumors and viruses in relation to diagnostic pathology. It also describes the use of conventional staining method such as negative stain and recently developed immunocytochemistry with colloidal gold labelling significantly improved diagnostic value of electron microscopy. The examples have been chosen to demonstrate quantitative aspects in the early recognition of immunopathological states, morphological aspects in identification of both tumors and viruses, preparative techniques of specimens where the method has been widely used and immunogold labelling, including various colloidal gold systems to map specific ligands. PMID- 8711881 TI - [The journals Acta Veterinaria (Brno), Veterinarni Medicina (Prague) and Czech veterinary sciencse 1960 to 1994]. PMID- 8711882 TI - Leptospirosis in farm animals. PMID- 8711883 TI - Reproductive performance of dairy herds infected with Leptospira interrogans serovar hardjo relative to the year of diagnosis. AB - To assess the impact of Leptospira interrogans serovar hardjo infection on the reproductive performance of nine dairy herds with evidence of infection, forty years' fertility data were analysed relative to the year of first diagnosis. Fifty per cent of various fertility variables had their lowest values only in the year of diagnosis. Culling rates were highest during the year of diagnosis in five of the herds, and were above 22 per cent in five of nine (55-6 per cent) of the diagnosis years considered compared with seven (22-6 per cent) of the 31 non diagnosis years. An assessment of the fertility status of the herds by means of a formula with incorporated the first service conception rate, the number of services per conception for cows conceiving, the calving to conception interval and the culling rate, revealed low reproductive performance during the year of diagnosis in six of the nine herds. Abortion rates were highest in four of the herds during the year of diagnosis, and these included the herds in which the fertility status was not lowest. It was concluded that L. interrogans serovar hardjo affected reproductive adversely by causing both abortions and low fertility, but that the effect was temporary. PMID- 8711884 TI - Use of polymerase chain reaction in the diagnosis of tuberculosis in cats and dogs. AB - Samples from four dogs and four cats suspected of having tuberculosis were processed for histopathology, bacterial culture and polymerase chain reaction (PCR). A simple, rapid method for the extraction of DNA from tissue samples was used in two PCR assays designed to confirm the diagnosis of tuberculosis. The PCR assays detected all the culture-positive samples from these animals and no false positive results were obtained. The PCR technique was successful for the direct detection of organisms from the Mycobacterium tuberculosis complex and reduced the time needed for a diagnosis to two days. PMID- 8711885 TI - Effect of salinomycin in the control of Clostridium perfringens type C infections in sucklings pigs. AB - The ability of salinomycin to control Clostridium perfringens type C infection in sows and their offspring was examined under field conditions. Two groups of sows and their offspring were offered feed either medicated with 60 ppm salinomycin or free of antibiotics, and their performance was compared. The number of piglets with diarrhoea, the duration of the diarrhoea, and the mortality of the piglets during the lactation period were markedly lower in the group given salinomycin. In addition, laboratory examinations showed that the numbers of carrier piglets and sows were reduced after treatment with Salinomycin. Finally, the sows treated with salinomycin lost less weight during the lactation period and weaned more and heavier piglets than the untreated sows. It was concluded that salinomycin incorporated in the diet can be used for controlling C perfringens type C infection in sows and their offspring. PMID- 8711886 TI - Evidence of torovirus infection in diarrhoeic cattle. PMID- 8711887 TI - Perinatal calf deaths. PMID- 8711888 TI - Wild Mammals (Protection) Act. PMID- 8711889 TI - Origin of BSE. PMID- 8711890 TI - Sensitivity of Serpulina hyodysenteriae to tiamulin in the UK. PMID- 8711891 TI - TB testing. PMID- 8711892 TI - Validity of serum bile acids for portocaval shunt screening. PMID- 8711893 TI - Fusobacterium necrophorum infections: virulence factors, pathogenic mechanism and control measures. AB - Fusobacterium necrophorum, a Gram-negative, non-spore-forming anaerobe, is a normal inhabitant of the alimentary tract of animals and humans. Two types of F. necrophorum, subspecies necrophorum (biotype A) and funduliforme (biotype B), have been recognized, which differ morphologically, biochemically, and biologically. The organism is an opportunistic pathogen that causes numerous necrotic conditions (necrobacillosis) such as bovine hepatic abscesses, ruminant foot abscesses and human oral infections. The pathogenic mechanism of F. necrophorum is complex and not well defined. Several toxins, such as leukotoxin, endotoxin, haemolysin, haemagglutinin and adhesin, have been implicated as virulence factors. Among these, leukotoxin and endotoxin are believed to be more important than other toxins in overcoming the host's defence mechanisms to establish the infection. F. necrophorum is encountered frequently in mixed infections and, therefore, synergisms between F. necrophorum and other pathogens may play an important role in infection. Several investigators have attempted to induce protective immunity against F. necrophorum using bacterins, toxoids, and other cytoplasmic components. Generally, none of the immunogens has afforded satisfactory protection against Fusobacterium infections. Because of the unavailability of suitable immunoprophylaxis, the control of F. necrophorum infection has depended mainly on the use of antimicrobial compounds. PMID- 8711894 TI - A comparison of the potency of several Brucella allergens used to detect brucellosis in cattle. AB - The potency of Brucella allergens prepared from a smooth Brucella abortus strain S-99, mucoid strain Leewarden, rough strain 45/20, and rough Brucella melitensis strain B-115 was assessed. The potency of these allergens was compared with that of a standard allergen prepared from smooth Brucella abortus S-99 that efficiently detected bovine brucellosis in other studies. Eight cattle experimentally inoculated with Brucella abortus 544 were tested with the allergens 4 and 10 weeks after infection, and again 8 months after infection. All the allergens effectively detected infection but there was a clear distinction in the mean skin reactions 48 and 72 h after injection of the allergens. The skin reactions provoked by the allergens prepared from smooth or mucoid strains of Brucella were most pronounced 48 h after injection. Skin reactions provoked by allergens prepared from rough strains of Brucella were strongest 72 h after injection. Allergens prepared from smooth or mucoid Brucella strains were more potent in detecting brucellosis than those prepared from rough strains of Brucella. PMID- 8711895 TI - Prevalence of agglutinating antibodies to Toxoplasma gondii in small ruminants of the Madrid region, Spain, and identification of factors influencing seropositivity by multivariate analysis. AB - A seroepidemiological survey of Toxoplasma gondii infection in sheep and goats was conducted in the Madrid region of Spain. Sera were collected from 60 herds, for which farming management information and other relevant data for their characterization were also obtained through a questionnaire. The seroprevalence was 11.8% (64 out of 541), using the modified (2-mercaptoethanol) direct agglutination technique with a 1:64 cut-off titre. The relationship between seropositivity and the variables in the questionnaire was assessed by multivariate analysis. Four variables were found to be significantly associated with seroprevalence. Two of them, the presence of cats and a previous history of abortion outbreaks in the farm, were factors known to be linked with toxoplasmosis, indicating the validity of the serological data. Seropositivity was also related to a lack of replacements in the preceding year. Proximity to other farms appeared to be a protective factor negatively associated with seropositivity, probably because it was an indicator of proximity to an urban area and the availability of local sanitary facilities. PMID- 8711896 TI - Isolation of keratinophilic fungi from the floors of private veterinary clinics in Italy. AB - To evaluate the presence of keratinophilic fungi in the environment, 400 samples were collected from the floors of 50 private veterinary clinics using 55-mm diameter 'contact plates', containing mycobiotic agar. After incubation for 15 days at 25 degrees C, the following species were isolated: Microsporum canis, Trichophyton terrestre, Chrysosporium keratinophilum, Chrysosporium sp., Microsporum gypseum, Trichophyton ajelloi, Chrysosporium tropicum, Trichophyton mentagrophytes, Chrysosporium state of Arthroderma tuberculatum and Chrysosporium pannorum. It is concluded that the keratinic material shed by infected pets may contribute to the development and propagation of dermatophytes and related fungi in veterinary clinics. Therefore, such veterinary clinics may represent sites where pets and humans are exposed to risk of infection with keratinophilic fungi from the environment. PMID- 8711897 TI - Residues of dimetridazole in eggs after treatment of laying hens. AB - Laying hens were dosed orally with dimetridazole (DMZ) (50 and 250 mg/kg) for 3 days or intramuscularly (50 mg/kg), also for 3 days, and the residues were determined by liquid chromatography in albumen and yolk. The sensitivity of the whole procedure was 2 ng/g. The drug was excreted preferentially into the yolk (about 57% of the total) and the elimination period lasted for 4-6 days after treatment. PMID- 8711898 TI - Depletion of the residues of colistin and amoxicillin in turkeys following simultaneous subcutaneous administration. AB - The tissue distribution and depletion of colistin and amoxicillin were studied in 84 turkeys dosed subcutaneously on 4 consecutive days with a formulation containing the two drugs at 0.2 ml/kg per day, corresponding to 50 000 IU of colistin sulphate/kg and 20 mg of amoxicillin trihydrate/kg. All the turkeys were killed 1-30 days after the final dose and samples of muscle, liver, kidney and cutaneous-subcutaneous tissues and of the injection site were taken for analysis for colistin and amoxicillin residues. The colistin concentrations in the liver (117.5 +/- 26.0 ng/g) and cutaneous-subcutaneous tissue (100.0 +/- 35.6 ng/g) were higher than those in kidney (92.0 +/- 34.4 ng/g) or muscle (67.5 +/- 16.9 ng/g) 1 day after the final dose. The concentration of this drug then increased for 9-14 days, followed by a slow decrease. The antibiotic was still present at low concentrations in the kidneys of all the treated birds and in the livers of two turkeys 30 days after the end of treatment. Amoxicillin concentrations were greatest in muscle (389.2 +/- 195.0 ng/g) and at the injection sites (440.3 +/- 213.9 ng/g) 1 day after treatment ceased, with a subsequent rapid decline. This drug was undetectable in the livers and kidneys by 10 days after dosing ceased. PMID- 8711900 TI - Preliminary observations on the efficacy of an iodophor in reducing the mortality in chickens experimentally affected by the 'hydropericardium syndrome'. PMID- 8711899 TI - The effects of glibenclamide, a blocker of K+ ATP-sensitive potassium channels, on diaphragmatic fatigue during endotoxaemia in pigs. AB - An in vivo porcine model of endotoxaemia was used to study the effects of glibenclamide, a K+ ATP-sensitive potassium channel blocker. Escherichia coli lipopolysaccharides (LPS, 70 micrograms/kg, i.v., as a bolus) were infused into anaesthetized, mechanically ventilated, indomethacin-treated pigs. After 120 min of endotoxaemia, glibenclamide was administered (10 mg/kg, i.v., over 5 min) to half the pigs. The strength at different frequencies of stimulation (10, 20, 30, 50 Hz, 20 V,) 1 s) and the endurance capacity (10 Hz, 20 V, 30 s) of the diaphragm were evaluated after 120 min of endotoxaemia and 5, 10, 20 and 30 min after drug infusion. Glibenclamide transiently increased the blood pressure without changing the decreased cardiac output and at the same time further impaired the diaphragmatic activity. The reduced ability of the diaphragm to generate force in response to different electrical stimulations was shown by a significant reduction in strength. The endurance index decreased 5 min after glibenclamide infusion, returning to the pre-glibenclamide values by 150 min. These results indicate that glibenclamide modifies the activity of vascular smooth muscle and of the diaphragm. PMID- 8711901 TI - Spectral tuning and molecular evolution of rod visual pigments in the species flock of cottoid fish in Lake Baikal. AB - Lake Baikal in Eastern Siberia is the deepest and one of the largest and most ancient lakes in the world. However, even in the deepest regions, oxygenation levels do not fall below 75-80% of the surface levels. This has enabled a remarkable flock of largely endemic teleost fish of the sub-order Cottoidei to colonize all depth habitats. We have previously shown that species that occupy progressively deeper habitats show a blue shift in the peak wavelength of absorbance (lambda max) of both their rod and cone visual pigments; for the rod pigments, a number of stepwise shifts occur from about 516 nm in littoral species to about 484 nm in abyssal species. By sequencing the rod opsin gene from 11 species of Baikal cottoids that include representatives from all depth habitats, we have been able to identify four amino acid substitutions that would account for these shifts. The effect of each substitution on lambda max is approximately additive and each corresponds to a particular lineage of evolution. PMID- 8711902 TI - No binocular rivalry in the LGN of alert macaque monkeys. AB - Orthogonal drifting gratings were presented binocularly to alert macaque monkeys in an attempt to find neural correlates of binocular rivalry. Gratings were centered over lateral geniculate nucleus (LGN) receptive fields and the corresponding points for the opposite eye. The only task of the monkey was to fixate. We found no difference between the responses of LGN neurons under rivalrous and nonrivalrous conditions, as determined by examining the ratios of their respective power spectra. There was, however, a curious "temporal afterimage" effect in which cell responses continued to be modulated at the drift frequency of the grating for several seconds after the grating disappeared. PMID- 8711903 TI - Coherence, cardinal directions and higher-order mechanisms. AB - Our initial purpose was to develop a quantitative method of estimating the cardinal directions of color space. The method is based on the finding that patterns consisting of pairs of drifting gratings modulated along different cardinal axes appear to slip with respect to one another, while the same patterns appear as a single coherent plaid if the modulation directions of the patterns are rotated by 45 deg in color space [Krauskopf & Farell (1990). Nature, 348,328 331]. A forced-choice procedure was used in which observers were asked to choose which of two successively presented patterns appeared less coherent. The patterns consisted of pairs of drifting gratings; the direction of modulation of one of the gratings was fixed and that of the other varied. For example, an estimate of an individual's isoluminant plane could be obtained by fixing the modulation of one grating in the luminance direction and finding the elevation of the modulation of the other grating that resulted in minimum perceived coherence. We found it important to take into consideration individual differences in the tilt of the isoluminant plane in color space and in the detectability of targets in the nominal cardinal directions. When this was done we found that reliable measurements could be made. The method effectively provided quantitative estimates of the cardinal directions. However, the most important result was the inadequacy of the generalization that patterns appear coherent when they share similar components along cardinal directions (Krauskopf & Farell, 1990) to account for the new results. The present results suggest that patterns appear not to cohere to the extent that they fail to stimulate common chromatic mechanisms, but the assumption that these mechanisms are tuned only along cardinal axes can be rejected. Along with other data the results point to the existence of higher order mechanisms tuned to different isoluminant chromatic directions. PMID- 8711904 TI - Further evidence for monocular determinants of perceived plaid direction. AB - This report adds to existing evidence that a monocular, feature-sensitive motion mechanism is involved in two-dimensional (2-D) motion processing, and also accounts for an earlier, unexplained result [Alais et al.(1994) Vision Research, 34, 1823-1834]. The central finding is that the perceived direction of a monocularly viewed type II plaid changes over a period of continuous exposure such that post-adaptation direction judgements exhibit more of the component direction bias known to occur with these stimuli than pre-adaptation judgements. These adaptation effects are confined to the adapted eye: when the adapting stimulus is presented to one eye, pre- and post-adaptation direction judgements made with the other, non-adapted eye are identical. These results strongly suggest the involvement of a monocular motion mechanism in two-dimensional motion processing, in addition to the more commonly presumed binocular mechanisms. PMID- 8711905 TI - The effect of display size on disparity scaling from differential perspective and vergence cues. AB - The present study compared the relative effectiveness of differential perspective and vergence angle manipulations in scaling depth from horizontal disparities. When differential perspective and vergence angle were manipulated together (to simulate a range of different viewing distances from 28 cm to infinity), approximately 35% of the scaling required for complete depth constancy was obtained. When manipulated separately the relative influence of each cue depended crucially on the size of the visual display. Differential perspective was only effective when the display size was sufficiently large (i.e., greater than 20 deg) whereas the influence of vergence angle, although evident at each display size, was greatest in the smaller displays. For each display size the independent effects of the two cues were approximately additive. Perceived size (and two dimensional spacing of elements) was also affected by manipulations of differential perspective and vergence. These results confirm that both differential perspective and vergence are effective in scaling the perceived two dimensional size of elements and the perceived depth from horizontal disparities. They also show that the effect of the two cues in combination is approximately equal to the sum of their individual effects. PMID- 8711906 TI - Contour integration with colour and luminance contrast. AB - In this study, we consider how colour contrast can be used to integrate form and how it interacts with luminance contrast in the task. The performance of form integration was assessed by measuring the detection of a winding "contour" of aligned gabor elements embedded in a background of randomly oriented gabors, using both luminance and isoluminant (red/green) contrast. Performance on the task improves with gabor element contrast, and identical performance for colour and luminance contour detection is achieved at high screen contrasts, showing that colour is able to support a complex form integration task. In a second experiment, we investigate whether colour and luminance contrast can be combined in contour integration by measuring the detection of a path with alternating isoluminant colour and luminance elements. We find that contour detection uses both colour and luminance information cooperatively, but performance is much poorer than would be expected from a single common contour integration process which fails to distinguish the two types of contrast. This suggests that there are specific contour integration processes for colour and luminance. In a third experiment, we measure the effects of variations in colour and luminance contrast on contour detection using elements that combine colour and luminance contrast. We find that varying the colour contrast of elements tends to worsen the detection of a luminance contour, as do luminance contrast variations for colour contour detection. These results suggest no special role for colour in integrating contours, and are discussed with regard to their ecological significance. PMID- 8711907 TI - Perceived velocity of luminance, chromatic and non-fourier stimuli: influence of contrast and temporal frequency. AB - We measured perceived velocity as a function of contrast for luminance and isoluminant sinusoidal gratings, luminance and isoluminant plaids, and second order, amplitude-modulated, drift-balanced stimuli. For all types of stimuli perceived velocity was contrast-invariant for fast moving patterns at or above 4 deg/sec. For slowly moving stimuli the log of perceived velocity was a linear function of the log of the contrast. The slope of this perceived velocity-vs contrast line (velocity gain) was relatively shallow for luminance gratings and luminance plaids, but was steep for isoluminant gratings and isoluminant plaids, as well as for drift-balanced stimuli. Independent variation of spatial and temporal frequency showed that these variables, and not velocity alone, determine the velocity gain. Overall, the results indicate that slow moving stimuli defined by chromaticity or by second-order statistics are processed in a different manner from luminance defined stimuli. We propose that there are a number of independent mechanisms processing motion targets and it is the interplay of these mechanisms that is responsible for the final percept. PMID- 8711908 TI - The interpretation of visual motion: evidence for surface segmentation mechanisms. AB - The independent motions of objects in a visual scene are commonly manifest as overlapping retinal motions. A consequence of this overlap is the creation of spurious retinal image features--such as corners and terminated contours--that bear no direct relation to the motions of the objects that give rise to them. To reconstruct object motions, these emergent features must be distinguished from the retinal motions of real object features. This process can be studied using visual stimuli known as plaid patterns, which provide a laboratory archetype for the ubiquitous real-world circumstance of two surfaces with overlapping retinal projections. By adjusting luminance relationships in a plaid pattern it is possible to influence the perceptual interpretation of image features, such that they are seen as either an emergent consequence of occlusion or as real variations in surface reflectance. In the former case, the plaid is most likely to be to perceived as two independently moving surfaces, whereas the latter generally elicits a percept of a single moving surface. This dependence of motion perception on luminance configuration can be viewed as evidence for the involvement of surface segmentation mechanisms, which distinguish between real and emergent image features by promoting a depth-ordered neural representation of surfaces. An alternative interpretation, which does not demand such depth ordering and feature classification, asserts that the effect of luminance configuration can be accounted for by attendant variations in the distribution of moving Fourier components. To evaluate these two proposed mechanisms, we designed novel plaid stimuli in which surface segmentation cues could be varied independently of changes in the distribution of Fourier components. Perceived motion was found to be highly correlated with the presence of appropriate segmentation cues and uncorrelated with the distribution of Fourier components. These results refute the Fourier components hypothesis, and they support our proposal that surface segmentation plays a critical role in the interpretation of visual motion signals. PMID- 8711909 TI - Stereoacuity and colour contrast. AB - We have measured the contrast dependence of stereoacuity using both horizontally and vertically oriented, isoluminant (red-green) and isochromatic (yellow-black), 0.5 c/deg Gabor patches. For comparison, contrasts were computed in multiples of detection threshold, where detection threshold was defined as the contrast required for the stimulus to be simultaneously detectable in each eye. Disparity thresholds (1/stereoacuity) for vertical chromatic Gabors were higher than those for vertical luminance Gabors by a factor of between 4 and 9 depending on contrast, and declined less steeply with contrast. Disparity thresholds for horizontal chromatic Gabors were very high (130-210 min arc) compared with horizontal luminance Gabors (by a factor of between 9 and 17) and were only measurable at contrasts above 10 times simultaneous monocular detection threshold. These results support the view that chromatic stereoscopic processing is less precise than luminance stereoscopic processing, and that there is a special deficit in the processing of disparity with horizontally oriented chromatic stimuli. The implications of these results for the role of colour vision in stereopsis are discussed. PMID- 8711910 TI - FPL and VEP measures of fusion, stereopsis and stereoacuity in normal infants. AB - Dynamic random dot fusion, stereopsis and stereoacuity were evaluated in 149 healthy, fullterm infants, using both forced-choice preferential looking (FPL) and steady-state visual evoked potential (VEP) protocols. Few infants aged 2-3 months demonstrated fusion or stereopsis in either the FPL or VEP protocol; most infants aged 5 months and older demonstrated fusion and stereopsis in both protocols. Both FLP and VEP stereoacuity approached adult-level (< 60 sec) by 6-7 months of age. Both infants and adults exhibited non-monotonic VEP amplitude vs disparity functions with a step change in phase at an intermediate disparity, consistent with separate fine and coarse disparity mechanisms. PMID- 8711911 TI - Hyperacuity for spatial localization of contrast-modulated patterns. AB - The acuity for localizing the position of a grating and other first order patterns which are defined directly by the luminance distribution, is much higher than the resolution for such gratings. This well-described phenomenon usually is referred to as hyperacuity, and is regarded as a cortical function which is not limited by the optics and the sampling properties of the eye. Second order patterns which can be defined by the distribution of local contrast gained some interest because they require more complex processing mechanisms than first order patterns. We investigated how well gratings and bars which are exclusively defined by the variation of the local contrast of static random dot patterns can be localized in space. In this case localization acuity does not reach the precision which is known for first order patterns. However, the localization of contrast-modulated patterns can be almost one order of magnitude better than second order grating resolution, and therefore reaches into the hyperacuity range. In combination with findings for motion-defined or stereo-defined patterns it is concluded that the brain mechanisms responsible for the localization of features in the visual scene have not only access to first order information which is available immediately from the retinal image, but in addition, to second order information which has to be extracted from the retinal intensity distribution by some sort of nonlinear processing. PMID- 8711912 TI - Anticipatory smooth eye movements of high velocity triggered by large target steps: normal performance and effect of cerebellar degeneration. AB - We studied frequencies and dynamic characteristics of anticipatory smooth eye movements (ASEM) in humans who were tracking step target movements of 20-70 deg amplitude. During presentation of periodic steps of constant amplitude healthy subjects showed frequent high velocity ASEM reaching maximal peak velocities of 5 40 deg/sec. There was no effect of ASEM on the frequency of anticipatory saccades. Randomization of target step amplitude or onset reduced the frequency of ASEM but did not completely abolish fast ASEM. In patients with cerebellar degeneration who exhibited impaired smooth pursuit, fast ASEM were absent and the number of slow ASEM was minimal. In conclusion, large sequential target steps can elicit much higher ASEM velocities than typically described in the literature. Similar to slow ASEM triggered by small steps, these fast ASEM do not require specific training and are not canceled by unpredictable step target motion. However, fast ASEM depend on the intact function of the cerebellum which gives further evidence of their generation by the smooth pursuit oculomotor subsystem. PMID- 8711914 TI - [Dentogenic sources of infection in patients with awaiting heart valve replacement]. AB - The detection of all teeth with periapical pathological conditions and optimal treatment of the affected teeth are a part of the evaluation program for patients who are scheduled to undergo cardiac value replacement. 48 patients (50 value replacement operations) were evaluated for the incidence of periapical pathological conditions, as well as for the different kinds of oral surgical interventions. A periapical pathological condition was present in 28 of 48 patients (58.3%). In general, the incidence of a dental focus was significantly higher in patients awaiting aortic valve replacement than in patients undergoing mitral valve replacement (69.7% vs. 23.1%; p < 0.01). Patients for aortic valve replacement presented significantly more periapical pathological conditions than patients for mitral valve replacement (1.7 vs. 0.8). Optimal timing of dental consultations is helpful in the oral rehabilitation of patients about to undergo a valve replacement procedure. Furthermore, close interdisciplinary cooperation is necessary to minimize the social and psychological problems following unexpected tooth extractions. Cardiologists and heart surgeons have an important role in the organisation of the oral rehabilitation of patients undergoing valve replacement procedures. PMID- 8711913 TI - [Diabetes and pregnancy]. AB - During the past years perinatal mortality in diabetic pregnancy has been lowered significantly, in special collections with highly motivated diabetic women even below 2%. In case of optimal metabolic control and absence of diabetic angiopathy the perinatal survival rate is identical to that in normal pregnancy. But adequate metabolic control cannot be reached in all pregnant women during the whole period of gestation, and pre-existing diabetic angiopathy exists frequently; therefore diabetic pregnancy will be associated with elevated perinatal mortality and morbidity also in the future. In case of pre-existing diabetic nephropathy, especially when GFR is decreased before conception there is a high risk for both progression of nephropathy during pregnancy and increased incidence of gestosis and intrauterine growth retardation which lead to earlier delivery (before the 36th week of gestation) in most cases. Perinatal mortality newborns of diabetic women with nephropathy has been decreased significantly during the past years, but perinatal morbidity in these newborns is still high. Pre-existing proliferative retinopathy can show progressive deterioration during pregnancy, but spontaneous regression post partum is usual. In rare cases progression of retinopathy also after delivery has been described even when laser coagulation was performed. Diabetic women with macroangiopathy, especially with coronary artery disease show a high risk for cardiovascular events during pregnancy. The occurrence of acute myocardial infarction in pregnant diabetic women is associated with elevated fetal and maternal mortality. Though there are case reports in the literature describing a successful fetal and maternal outcome after myocardial infarction during pregnancy. PMID- 8711915 TI - [Management of abdominal stab wounds]. AB - In general, laparotomy is accepted as treatment of choice for abdominal shot wounds. However, the management of abdominal stab wounds is controversial. In order to study the concept of laparotomy for penetrating abdominal trauma our patient collective was analyzed retrospectively. From 1st January 1985 to 31st December 1993 we performed laparotomy in 30 cases of abdominal stab wounds. 24 patients were victims of violence. In 6 patients the cause of injury was a suicidal attempt. Laparotomy was negative in 27% of cases. In most cases explorative laparotomy showed injuries of the vessels (n = 12) and the intestines (n = 9). The mortality rate was 6.7%. None of the fatal cases was caused by negative laparotomy. During a mean follow-up time of 54 months none of the patients showed disorders of late onset after negative laparotomy. Because of the limited diagnostic possibilities to rule out intestine injury and minimal morbidity of negative laparotomies we continue to favour laparotomy in the management of penetrating abdominal trauma. PMID- 8711916 TI - [Urinary incontinence in patients with Parkinson syndrome]. AB - Disturbances of micturition are very common in Parkinson's disease. Hence, we assessed 50 patients (22 female, 28 male, average age: 59.8 years) by means of the Gaudenz score. In 31 cases (62%) normal findings were obtained. Urge incontinence was diagnosed in 13 cases (26%) and stress incontinence in 5 patients (10%). One patient had both urge and stress incontinence. All patients with stress incontinence were female. PMID- 8711917 TI - [Development, stimulation and paralysis of embryonic motor activity]. PMID- 8711918 TI - [The shoulder joint--diagnostic imaging]. AB - In the assessment of shoulder joint abnormalities plain films must still be used today as primary imaging modality. For soft tissue, cartilage, tendon and ligament lesions, CT with arthrography and particularly MRI with arthrography is available today. Especially latter method leads to a significantly extended diagnosis, which in many cases implies a major therapeutical significance. PMID- 8711919 TI - [Ultrasound examination of the shoulder joint]. AB - The ultrasound examination of the shoulder is an effective, quick and painless diagnostic method. The skilled examiner has no problem to detect rotator cuff tears, bursitis, tendinitis and instabilities as well as arthrosis or hematoma of the acromioclavicular joint and biceps tendon rupture. In the literature the sensitivity of ultrasound examination of the shoulder depended from 91 to 100% and specificity from 83 to 100%. The dynamic examination gives a special view of gliding tendons, muscles and bony landmarks of the shoulder. With standardized pictures the orthopedic surgeon should be able to diagnose and document almost every shoulder soft tissue pathology. PMID- 8711920 TI - [Rheumatoid orthopedics and endoprosthetics of the shoulder joint]. AB - The first part of the present article gives a comprehensive description of the pathology, diagnostics and forms of therapy of rheumatically impaired shoulder joints. Improved operative methods and methods such as bursectomy, synovectomy, reconstructive soft-tissue management and joint replacement depending on the stage of impediment have enhanced the use of surgery in the treatment of rheumatic shoulder joints. The second part gives a critical evaluation of indications, methods and results of modern shoulder joint replacement. A survey of written publications on the results of hemi- and total endoprostheses on the one hand, our own clinical experiences on the other, have shown that the use of modular hemiendoprostheses produces clinically excellent results with no problems of loosening of the glenoid part. PMID- 8711921 TI - [Frequent injuries of the shoulder joint]. AB - This article tries to identify the most frequent osseous lesions in the shoulder complex found in the all day routine of a trauma center or trauma unit. It discusses the clinical diagnosis, the most typical radiological findings and the current concepts of treatment. This report is designed mainly to help the general practitioner outside the hospital community. PMID- 8711922 TI - [Rupture of the rotator cuff]. AB - The operative treatment of rotator cuff tears becomes more popular. New reliable diagnostic techniques improved operative procedures and as a very important fact the demanding of the patients to a normal function of the shoulder account for this development. Therefore every physician has to know exactly the reason, the clinical appearance and the course of this pathology. Beside conservative treatment, new operating techniques and the postoperative management will be shown. PMID- 8711923 TI - [Conservative therapy of shoulder pain]. AB - Prerequisite to conservative therapy for shoulder pain is the diagnosis. Along with the topical diagnosis (omarthralgia) the structure analysis aims to identify the site and manner of disturbed structure. Destroyed structures are, in most cases, an indication for anatomical reconstruction, and disturbed structures an indication for conservative methods. Hereby acute symptoms require pain stimulus pacification and chronic symptoms target-specific irritant application. With disorders of the muscles their insertions and the gliding tissue, and in addition to physical methods such as thermotherapy and electrotherapy, the so-called needle treatments (local anesthesia, combination with corticoids, needling, and acupuncture) are to be mentioned. Manual therapeutic methods will also be discussed. With disturbances of the joint function use of intra-articular injections is to be considered, with movement restriction the mobilisation techniques of chirotherapy. Physiotherapy exercises are of particular importance here. PMID- 8711924 TI - [Periarthritis humeroscapularis--clinical diagnosis and analysis of the syndrome concept]. AB - Based on the recent knowledge on shoulder pathology the diagnosis of periarthritis humeroscapularis is no longer considered to be adequate. In this article the different pathologies are presented. For all important clinical entities the clinical and functional tests are shown, which are helpful to differentiate the different pathologic features. PMID- 8711925 TI - [Arthroscopy and arthroscopic surgery of the shoulder joint]. AB - Shoulder arthroscopy is a valuable procedure in unclear shoulder pain and the differential diagnosis between instability and impingement. With arthroscopic subacromial decompression comparable results to open operations can be achieved whereas in stabilizing procedures of the shoulder joint, open surgery is still superior to arthroscopic operations. PMID- 8711927 TI - Age differences in attitudes toward menopause and estrogen replacement therapy. AB - The purpose of the present study was to develop and test attitude scales for menopause and estrogen replacement therapy (ERT) using 116 college-aged and 136 mid-aged women. Factor scores indicate that mid-aged women view menopause in a more benign fashion than college-aged women and are more likely to view ERT as positive while recognizing side effects. Restricted variability on ERT attitude items suggests limited knowledge or opinions and a need for education across ages. Women's perceptions of ERT are in terms of a solution for immediate relief of symptoms in contrast to current medical recommendations that emphasize ERT as long-term disease prevention therapy. PMID- 8711926 TI - [Diagnosis and therapy of shoulder instability]. AB - Shoulder instability is based on various acquired or constitutional aspects which determine the therapy and clinical outcome. In order to differentiate between traumatic and atraumatic forms of shoulder instability a careful history has to be taken. A systematic clinical examination is necessary to evaluate the shoulder joint for general laxity or true instability and especially to assess direction and degree of glenohumeral translation. The basic diagnostic imaging includes X rays in 3 planes. CT-arthrogram and MRI may provide additional information to evaluate the intraarticular pathology. An elective therapy depends on the patients age and demands. Recurrent anterior dislocations are treated by an anatomical reconstruction of the glenohumeral ligament complex. Open or arthroscopic Bankart-repair have been established as the treatment of choice. Conservative treatment is primarily indicated in cases of atraumatic posterior or multidirectional instability. If physiotherapy fails, surgical treatment should be discussed for these patients. PMID- 8711928 TI - Women living with HIV: sexual behaviors and counseling experiences. AB - Seventy-two ethnically diverse women attending two outpatient HIV clinics in Los Angeles self-administered questionnaires. Nineteen women (26%) reported sexual activity in the two months prior to the survey; most were sexually monogamous and emotionally involved with live-in male partners. According to self-reports, 13 women (18% of the total sample and 68% of those who were sexually active) had engaged in at least one form of risky sexual activity. The majority (81%) of sexual partners were aware of the respondent's HIV infection, but most (67%) were seronegative. Post-test counseling was reportedly noncomprehensive (e.g., 47% of the respondents did not recall receiving information about using condoms correctly). Few women accessed HIV support services. Results underscore the need for more effective counseling and support interventions for this population. PMID- 8711929 TI - The relationship between smoking, cholesterol, and HDL-C levels in adult women. AB - The purpose of this study was to determine the independent relationship between smoking quantity and cholesterol (TC) and lipoprotein levels (HDL-C) in women. A total of 805 female subjects were grouped as: non-smokers, ex-smokers, light smokers, moderate smokers, and heavy smokers. TC and HDL-C were examined before and after controlling for the coexisting risk factors of age, body composition, fitness level, dietary fat intake, family history of coronary artery disease, oral contraceptive, and alcohol use. Preliminary analysis demonstrated significant differences (p < .01) in TC between heavy smokers and all other groups and significant differences in HDL-C between heavy to moderate smokers and ex- and non-smokers. After adjusting for confounding variables, the differences in TC and HDL-C remained unchanged between the groups. It was concluded that heavy to moderate smoking was independently associated with higher TC and lower HDL-C levels, and that smoking abstinence or smoking cessation may be associated with healthier lipoprotein profiles in adult women. Based on these findings, it was recommended that employers consider the provision of health promotion programs including seminars, behavioral modification workshops, as well as financial incentives for employees to stop smoking. PMID- 8711930 TI - Cultural differences in attitudes, values, and beliefs about osteoporosis in first and second generation Japanese-American women. AB - This study examines attitudinal differences related to osteoporosis between first and second generation Japanese-American women. In an interview, the women completed a battery of tests assessing their attitudes, values, and beliefs about the diagnosis, treatment, and follow-up care of osteoporosis. The groups differed in their general knowledge of osteoporosis, perceptions of the disease, attributions of its causes, anticipated and preferred support mechanisms for care, and anticipated areas of concern for self-or other-care. There were also considerable differences in treatment compliance and feelings toward physicians. The findings were discussed in relation to the effects of culture on health-care attitudes and behaviors. PMID- 8711931 TI - Feedback for participants in a health survey: feasible and useful. AB - The use of quantitative methods has often limited the communication between researchers and research participants to being one-sided. Use of feedback in survey-oriented research is one possibility for creating a more communicative relationship. After doing a postal survey of 2000 Finnish 45 to 64-year-old women about their climacterium we produced a feedback leaflet about the main results of the study and about climacterium in general and sent it to all respondents (n = 1713). Later a postal questionnaire concerning the feedback leaflet was sent to a consecutive sample of every eighth (n = 200) woman, of whom 153 (76%) responded. Most of the women gave neutral or positive responses to the content of the leaflet. There was more interest in the feedback among the younger and pre menopausal groups than among the older or postmenopausal women. Some women wanted more information about climacterium-related issues than was possible to give in the space of eight pages. According to this experiment, feedback offers fresh view-points and valuable critique for researchers as a result of being more in contact with respondents. PMID- 8711932 TI - Iron related indices in iron deficiency anemia of geriatric Korean patients. AB - The purpose of this study was to compare iron related indices in patients with iron deficiency anemia and chronic causative diseases between geriatric older than 65 years and adult, nongeriatric younger than 65 years groups. Iron deficiency anemia (IDA) cases with chronic disorders from Youngdong Severance hospital from June, 1991 to April, 1994, older than 65 years (17 cases), and younger than 65 years (29 cases) were analysed with iron related indices. Mean hemoglobin was 7.8 +/- 2.2 g/dl in geriatric IDA and 8.0 +/- 1.8 g/dl in adult IDA without significant difference. RDW value was 19.5 +/- 2.6 in geriatric IDA and 18.4 +/- 3.2 in adult IDA with no significant difference. Serum iron and transferrin saturation between geriatric IDA were 22.7 +/- 12.3 ug/dl, 6.7 +/- 4.1% and 28.6 +/- 16.6 ug/dl, 7.1 +/- 4.4% in adult IDA with no significant difference, but TIBC was significantly lower (P = 0.011) in geriatric IDA than in adult IDA patients (357.2 +/- 83.2, 413.6 +/- 54.0 ug/dl). In normal elderly people, serum ferritin was 152.5 +/- 95.4 ng/ml in male and 111.1 +/- 54.1 ng/ml in female with range 19.8 approximately 367.7 ng/ml in male and 11.7 approximately 238.7 ng/ml in female and was higher than that of normal adult in both sexes (147.0 +/- 108.0, 35.3 +/- 20.5 ng/ml) (P = 0.045). Serum ferritin in geriatric IDA was 13.8 +/- 11.8 ng/ml and 5.7 +/- 4.0 ng/ml in adult IDA with significant difference(P = 0.001). The Upper margin for geriatric IDA was 37 ng/ml with 95% confidence interval. In the diagnosis of geriatric IDA with causative diseases, we should consider that TIBC does not increase and the upper margin for serum ferritin is suggested to increases up to 37 ng/ml. PMID- 8711933 TI - p53 protein expression in extrahepatic bile duct cancer. AB - p53 mutations, a tumor suppressor gene located on chromosome 17p, are the most common genetic alterations found in human cancers. Although the p53 expression or mutation has been investigated in a variety of cancers there have been very few studies in extrahepatic bile duct cancers. In this study, we investigated the immunohistochemical expression of p53 in formalin fixed paraffin embedded archival specimens of 36 extrahepatic bile duct cancers in which p53 expression was found in eighteen (50%) cases. There was no significant difference in age, gender, size of tumor, histologic grade, extent of tumor involvement, lymph node metastasis and tumor resectability according to p53 immunoreactivity. Comparison of survival duration according to p53 expression showed no significant difference. In conclusion, we reported 50 percent of p53 expression in extrahepatic bile duct cancers by immunohistochemical staining and we found no prognostic significance of p53 expression in dinicopathologic parameters. PMID- 8711934 TI - The role of omentopexy in tracheal transplantation in dogs. AB - The major step toward successful tracheal transplantation is revascularization of the grafted trachea. There are many reports that although omentopexy is an effective method to facilitate neo-vascularization in tracheal transplantations, the procedure has not been accepted universally in the transplantation field. It remains unclear whether an omentopexy can successfully revascularize tracheal graft regardless of the length of graft. This study was undertaken to assess the usefulness of omentopexy for long-segment(more than 4 cm) tracheal allotransplantation. We have performed six tracheal transplantations with omentopexy (group A) and four tracheal transplantations without omentopexy (group B) in mongrel dogs from July 1993 to February 1995. Five mid-portion tracheal rings were removed from ten donor dogs and ten corresponding tracheal rings were removed from the ten recipient dogs. The excised tracheal rings from the donors were transplanted to the recipient tracheal-excised sites. All the recipients were given cyclosporine, azathioprine, and prednisolone for immunosuppression in the post-operative period. The histologic results of all the surviving members of group B were better than those of the group A. These findings indicate that omentopexy has a limitation, it is not a major method for graft revascularization. Therefore the length of the tracheal graft was greater than 4.0 cm, for its viability, a longer tracheal graft requires some other blood supply aside from the omentopexy. PMID- 8711935 TI - Increased serum interleukin-10 level in Kawasaki disease. AB - Serum IL-10 level in Kawasaki disease(KD) was tested. In the KD patients' sera during the acute phase, the levels of IL-10 were markedly elevated (122.0 +/- 39.1 pg/ml) compared to 3.7 +/- 1.7 pg/ml in the control subjects (p < 0.001). The serum IL-10 levels remained elevated in the subacute phase (16. 7 +/- 9.7 pg/ml, p < 0.001) and were restored to the normal levels(7.9 +/- 3.9 pg/ml) during the convalescent phase. In the patients with acute febrile disease, the serum IL-10 level increased significantly (34.4 +/- 14.1 pg/ml, p < 0.001) compared to that of the age-matched control subjects, but were not as high as in acute phase of KD(p < 0.005). This increase in serum IL-10 levels in KD may contribute to the up-regulation of humoral immunity and to the down-regulation of acute inflammation due to an increase in proinflammatory cytokines. PMID- 8711936 TI - Solid and papillary neoplasms of the pancreas. AB - Solid and papillary neoplasms of the pancreas, a rare tumor usually found in young female patients, seldom presents with metastasis since it is a tumor with low potential for malignancy. The prognosis for this lesion is much more favorable than that for other pancreatic neoplasms. In an attempt to understand the characteristics and prognosis of this lesion, we reviewed twenty cases treated at the Department of Surgery, Severance Hospital, Yonsei University from 1985 to 1994. The mean age of the patients was 25.6 years (range: 13 to 39 years), and 19 (95%) were women. Chief complaints were palpable mass (50%), pain (45%), and indigestion (5%). In laboratory studies, tumor markers, including CEA, CA125, CA19-9, and aFP were studied in eight patients, and found negative. Other laboratory findings were also nonspecific. These tumors may occur anywhere in the pancreas. In our studies, the tumor was most often located in the tail (45%), and the head (40%) of the pancreas. These were treated by distal pancreatectomy and splenectomy (55%), Whipple's operation (20%), pylorus preserving pancreatoduodenectomy (10%), enucleation (10%) or excision (5%). Significant morbidity or mortality was not observed during hospitalization, and no recurrence or malignant degeneration occurred during the mean follow-up period of 4 years (range: 1 month to 9 years). In conclusion, this study has suggested that the patients with a solid and papillary neoplasm of the pancreas have a good prognosis for successful treatment, if the disease is diagnosed early and the tumor is completely resected. A higher index of suspicion, and more aggressive diagnostic workups are needed in dealing with this disease entity. PMID- 8711937 TI - Cyclin D1 protein expression in lung cancer. AB - Cyclin D1, a G1 cyclin, has been implicated in the oncogenesis of various types of malignancies via deregulation of cell cycles. Amplification of cyclin D1 as a part of 11q13 amplicon has been reported in lung cancer as well as a subset of carcinomas arising from various organs including breast, head and neck, and esophagus. In addition to its role as an oncogene, several recent studies have suggested that amplification is indicative of poor prognosis. In this study we examined the cyclin D1 protein expression in 102 consecutive cases of lung cancers using the microwave enhanced immunohistochemical staining method and correlated the data with the histologic subtype and grade, Ki-67 (MIB-1) labeling index, and survival. Nuclear positive staining was observed in 18 cases (18 %) of lung cancers. Although squamous cell carcinoma demonstrated a higher rate of expression (12 /58, 21%), three of 33 adenocarcinomas (9%) revealed overexpression and both adenocarcinoma and squamous cell carcinoma components within the adenosquamous carcinoma showed nuclear staining. There was no correlation between cyclin D1 overexpression and histologic grade, Ki-67 (MIB-1) labeling index, and survival. These observations indicate that cyclin D1 protein overexpression might be implicated in the oncogenesis of the various histologic types of non-small cell lung carcinomas but it has no usefulness as a prognostic marker. PMID- 8711938 TI - A case of paraesophageal hernia repaired by laparoscopic approach. AB - Paraesophageal hernia comprises only 2 approximately 5% of all hiatal hernias but is prone to incarceration and strangulation. For this reason they must be recognized and repaired as expeditiously as possible. The laparoscopic approach has already been successfully applied to the repair of the more common sliding hiatal hernia and it seems reasonable to propose that the paraesophageal hernia, provided it is not complicated, might also be repaired by the laparoscopic technique. We present here a case of paraesophageal hernia which has been successfully repaired by the laparoscopic approach. A 73-year-old female suffering from postprandial fullness in the retrosternal area was diagnosed preoperatively with paraesophageal hiatal hernia with gastroesophageal acid reflux and was submitted for laparoscopic repair. The procedure entailed reduction of the hernia, mobilization of the esophagogastric junction with crural repair and partial fundoplication. At the 9th-month follow-up, the patient had remained asymptomatic and follow-up studies revealed no evidence of hernia or acid reflux. As a result of this favorable experience with minimal morbidity, early hospital discharge, and effective control of symptoms without adverse sequalae, laparoscopic repair can be considered as the curative and minimal invasive method in the management of paraesophageal hernia. PMID- 8711939 TI - Spontaneous bowel perforation during the course of acute pancreatitis--a case report. AB - We recently experienced a case of spontaneous perforation of infected necrosis into the colon and duodenum during the course of acute pancreatitis in a 63 year old male patient. Enteric perforations or fistulas in the setting of acute pancreatitis implicate severe underlying pathology and have substantial morbidity and mortality. In the meantime it has generally been accepted that infected pancreatic or peripancreatic necrosis should be managed surgically as soon as possible. Enteric perforations in the present case contributed to transient improvement of the patient's infection sign and condition, and thus an early operation was able to be avoided. Delayed surgical management resulted in complete recovery of the patient without postoperative morbidity. Herein we report an unusual complication of acute pancreatitis. PMID- 8711940 TI - The role of H1- and H2-receptors in the effect of compound 48/80 in the asphyxiation and body temperature of mice. AB - Contribution of histamine H1- and H2-receptors to the effect of compound 48/80, a potent histamine releaser, upon asphyxiation and body temperature in mice was investigated in the present experiments. Compound 48/80 showed an apparent protective potency against hypoxia and significantly prolonged the latencies for convulsions and death in a dose-dependent manner. Compound 48/80 also decreased the body temperature, which was in relation with the antihypoxic effect. Both the H1-receptor antagonist, dimethindene, and the H2-receptor antagonist, ranitidine, attenuated the hypothermic effect of compound 48/80, indicating the involvement of central histamine through both the H1- and H2-receptors. Ranitidine had no effect on the protective effect of compound 48/80 against hypoxia-induced lethality, whereas dimethindene completely antagonized it. These results suggest that the protective effect of compound 48/80 against hypoxia is mediated through histamine H1-receptors and is not related to its ability to induce hypothermia. PMID- 8711941 TI - [Coronary angiography in patients over 80 years of age. Therapeutic consequences and long-term follow-up]. AB - Today, an increasing number of old and very old patients is referred for invasive cardiovascular procedures. At our institution, cardiac catheterization has been performed in 82 patients 80 years of age or older (50 m, 32 f; mean age 82 +/- 2 years) during the last 11 years; in 70 patients, because of clinically proven or suspected coronary heart disease, and in 12 patients, because of heart valve disease. In comparison with younger patients, elderly patients with coronary heart disease more often presented with unstable angina, or had multivessel disease and reduced ejection fractions. PTCA (n = 25) and bypass grafting (n = 7) aimed at revascularization in 32 patients; in 38 patients, medical treatment was continued. Furthermore, valve replacement (n = 7) or valvuloplasty (n = 1) were judged to be indicated in 8 of the 12 patients with heart valve disease. Diagnostic procedure was more frequently burdened with serious complications in elderly than in younger patients (5/82 versus 3/300, p < 0.01). During follow-up of 25 +/- 23, median 21 months, cardiovascular events were significantly less frequent in patients with coronary heart disease who had undergone revascularization procedures than in those with medical therapy (2/29 versus 9/36; p = 0.05); more revascularized patients were free of angina at the time of reevaluation. Thus, risks of invasive diagnostic procedures are increased in very old patients. Diagnostics, however, result in therapeutical consequences, revascularization obviously improves symptomatology and prognosis in patients at extremely advanced age suffering from coronary heart disease. PMID- 8711942 TI - [Shock-induced, but not terminated ventricular tachycardia in a patient with implantable defibrillator]. AB - A 62-year-old male patient with coronary artery disease and drug refractory sustained ventricular tachycardia received an implantable cardioverter defibrillator (PRX III, model 1720, CPI) with a transvenous lead system (Endotak, model 0115, CPI) in combination with a subcutaneous array electrode (SQ array, model 049, CPI). The intraoperative defibrillation threshold was 15 J and was confirmed 1 week later at the hospital discharge test. Three months after discharge from hospital the patient reported 5 shocks during moderate physical exertion followed by a tachycardia associated with dizziness which terminated spontaneously. The print out of the stored electrogram showed a supraventricular tachycardia (probably sinus tachycardia) with a heart rate of 154/min which activated the device. Antitachycardia pacing did not terminate the supraventricular tachycardia, and hence shock therapy was delivered. The first shock (34 J) converted the supraventricular tachycardia to a ventricular tachycardia with a heart rate of 178/min, which was not terminated by four consecutive 34 J DC shocks. There was no hint of a device or lead failure. Determination of the defibrillation threshold reconfirmed the 15 J value for termination of ventricular fibrillation. However, neither a 1 J shock nor a 34 J shock terminated a monomorphic sustained ventricular tachycardia (cycle length 340 ms) induced by noninvasive programmed electrical stimulation. The ventricular tachycardia was, however, reproducibly terminate by antitachycardia pacing. It is concluded that an inappropriate high-energy DC shock might induce a sustained ventricular tachycardia. However, a sufficient defibrillation threshold for the termination of ventricular fibrillation does not necessarily mean that a sustained ventricular tachycardia will be terminated by a high-energy DC shock. PMID- 8711943 TI - ["Circumscribed defibrillator erythema": a differential diagnostic and therapeutic problem in differential occult defibrillator infection diagnosis]. AB - We report about three out of 452 patients (pts) (< 1 %) with circumscribed erythema (E) after implantation of an automatic cardioverter defibrillator (ICD). ICD-E occurred in two pts after pulse generator replacements and in one pt after initial ICD implantation. There were no clinical signs of inflammation and laboratory and technical finding were completely normal. Allergic reactions could be ruled out in all pts. Local or systemic drug treatment did not influence the ICD-E. During a follow-up of 28, 10 or 9 months, the ICD-E is still present in two patients and much improved in one pt without any treatment. We did not observe an infection in any of the pts. Our data show that the ICD-E occur rarely after ICD implantation and no special treatment is necessary if an infection or allergic reaction has been excluded. Nevertheless, it is sometimes difficult to exclude in those pts an atypical infection. PMID- 8711944 TI - [Calcium antagonists in treatment of ventricular tachycardia]. AB - Of the traditional antiarrhythmic agents administered in the treatment of ventricular tachycardias, those belonging to Class III are most commonly used. However, some ventricular tachycardias displaying special clinical, electrocardiographic and/or electrophysiologic characteristics have been successfully treated with calcium antagonists. Otherwise, Class IV antiarrhythmic agents are primarily used in the treatment of supraventricular tachyarrhythmias. It is reasonable to suspect that calcium-dependent phenomena underlie ventricular tachyarrhythmias in a number of patients. Verapamil-sensitive sustained left ventricular tachycardias represent a typical example. Sufficient response to calcium antagonists has also been demonstrated for exercise-induced ventricular tachycardias and repetitive monomorphic ventricular tachycardias. Characteristically, these arrhythmias occur in young patients with no structural heart disease. The long-term prognosis of these patients is in fact very good, but because of intrusive symptoms, pharmacological or nonpharmacological (radiofrequency ablation) treatment is often indicated. Through correct diagnosis and implementation of a short- and long-term therapy with calcium antagonists, and excellent alternative to the potentially harmful therapy with Class I and Class III antiarrhythmic agents can be offered. PMID- 8711945 TI - [In vivo registration of monophasic action potentials--new possible applications in clinical electrophysiology]. AB - Monophasic action potential (MAP) recordings have gained a more important role during the recent years. A new application for their use in the clinical laboratory is the precise differentiation between ventricular tachycardias and ventricular fibrillation. A correct diagnosis of the arrhythmic episode may be of relevance for the determination of the defibrillation threshold during implantation and testing of implantable cardioverter defibrillators. Diagnostic studies and ablation procedures of triggered arrhythmias represent another indication for MAP recordings. The focus of enhanced automaticity can be located by the detection of afterdepolarizations which allows a reliable ablation using a MAP-ablation combination catheter. Recently, a new phenomenon has been reported regarding the relationship between ventricular repolarization and excitability during programmed electrical stimulation in humans. Each additional extrastimulus was able to capture the myocardium at a less complete repolarization level than the previous one. This "facilitation of excitability" phenomenon or "encroachment" was correlated with the initiation of ventricular tachyarrhythmias. The involved mechanism can be modulated pharmacologically and might therefore play a role in the antiarrhythmic treatment of patients with ventricular arrhythmias. PMID- 8711946 TI - [Sex differences in the correlation between obesity and hypertension with left ventricular mass and hypertrophy]. AB - Overweight and hypertension are considered to be independent contributors to the development of left ventricular hypertrophy (LVH). We investigated a selected subgroup (n = 520, aged 52 to 67 years) of participants from the MONICA Augsburg cohort to assess gender-specificity of left ventricular adaptation in response to increasing weight and blood pressure degrees. M-mode-echocardiographic measurements were made and calculated according to the Penn-convention in 293 women and 227 men. LVH was defined as left ventricular mass indexed to height (LVMIm) > 143 g/m in men and > 102 g/m in women (Framingham criteria). Men and women were comparable with regard to increase in LVMIm from the lowest weight and blood pressure group to the highest groups, respectively. In men the increase in LVMIm was 31% from lean to severely obese subjects (111 vs. 145 g/m, p < 0.003) and 25% from normotensive to treated hypertensive subjects (116 vs. 145 g/m, p < 0.0001); in women respective values were 36% (83 vs. 113 g/m, p < 0.0001) and 27% (88 vs. 112 g/m, p < 0.0001). The combined occurrence of obesity and hypertension had an additional effect on left ventricular mass, which was much more pronounced in women than in men. In particular, the increase in LVMIm from the group of lean normotensives to the group of severely obese treated hypertensives was 85% (72 g/m vs. 133 g/m, p < 0.0001) in women and 49% (96 g/m vs. 144 g/m, p < 0.002) in men (p-value for the gender-interaction term < 0.05). The odds ratio for the LVH prevalence in hypertensive obese subjects as opposed to normal weight normotensive subjects were 11.9 (p < 0.0001) in women and 4.9 (p < 0.0004) in men. In conclusion, we observed for both genders independently and similarly pronounced effects of hypertension and obesity. The combined occurrence of hypertension and obesity had an additional impact on left ventricular mass and hypertrophy, however, in women the effects were significantly more pronounced than in men. The data underscore the effects of hypertension and obesity in the development of LVH. In addition, gender specific factors seem to modulate the effects of these risk factors on left ventricular mass. PMID- 8711947 TI - [Possible errors in diagnosis of left atrial thrombosis in transesophageal echocardiography]. AB - The detection of thrombi in the left atrium and left atrial appendage by transesophageal echocardiography has significant diagnostic and therapeutic value. However, it requires high method accuracy in routine evaluation. In order to record the percentage of false positive findings, a reevaluation was performed by two experienced examiners using the video recordings of 51 patients, in whom a thrombus has been diagnosed during the routine evaluation. From 1988 to 1994, these thrombi were diagnosed in 726 patients in whom we looked for a source of possible cardiac embolism. In the reevaluation of the 43 cases with sufficient video recordings, in 26 (60%) the diagnosis was classified as false-positive and in 5 patients (12%) as questionable. Thrombus diagnosis was confirmed in only 12 (28%) cases. The incidence of false-positive findings decreased over the course of years with 8.3% (1988-1990), 2.9% (1991-1992) and 0.4% (1993-1994), p = 0.0001. The agreement in the reevaluation between the two examiners was high (interobserver variability 7%), whereby completely divergent evaluations did not occur. Typical diagnostic pitfalls included a prominent trabecular structure in the left atrial appendage (n = 10), the membranous structure between the left atrial appendage and the upper left pulmonary vein (n = 7), transition of the lower left atrium/left atrial appendage (n = 3) and branches within the apex of the left atrial appendage (n = 2). When searching for thrombi in the left atrium and left atrial appendage, there is a high risk of false-positive findings, especially for the unexperienced investigator. Attention must be paid to the typical causes of diagnostic pitfalls in the case of complex and variable anatomy. PMID- 8711948 TI - [Post-traumatic aneurysma spurium of the right atrium]. AB - In a 61 year old male with heart failure and pulmonary congestion the x-ray shows a right paracardial tumor. The patient suffered from a blunt chest trauma 6 years ago. Since that accident he complains about exercise related dyspnea and cardiac arrhythmia with atrial fibrillation. On echocardiography we found a echolucent cystic tumor with a solid center structure surrounded by a thin membrane. Doppler echocardiography revealed a heart cycle dependent flow at its margin. During dextrocardiography rapid opacification only of the peripheral structures of the tumor could be observed. These findings are consistent with a traumatic rupture of the right atrium, and the diagnosis of a posttraumatic aneurysma spurium of the right atrium was established. Surgery confirmed this diagnosis and the aneurysm was extirpated. PMID- 8711949 TI - [Response to the letter by W. Forster on: prevention of reinfarction with 100 mg or 30 mg aspirin daily]. PMID- 8711950 TI - Applicability of neutron activation analysis (NAA) in quantitative determination of some essential and toxic trace elements in food articles. AB - Accurate and reliable data on microgram and nanogram quantities of some essential and toxic elements in most food articles are very scarce. Neutron activation analysis (NAA), with its essentially blankfree advantage, is a valuable approach in the field of determination of trace elements in different foodstuffs and diets. Accordingly, various radiochemical (RNAA) and instrumental (INAA) approaches have been developed in our laboratory for the element As, Cd, Co, Cu, Hg, I, Mo, Ni, Sb, Se, Sn, Th, U, V, and others, and verified by the analysis of compositionally appropriate certified reference materials. PMID- 8711951 TI - Detection of irradiated fruits by gas-chromatographic methods. AB - To detect those fruits which have been subjected to low-dose irradiation (0.5-3 kGy), two methods of chromatography (GC-MS and LC-LC-GC-FID) were used to determine the radiolytic compounds of lipids formed after irradiation, such as alkanes and alkenes. Extraction of volatile hydrocarbon compounds from some parts of irradiated fruits, e.g. the flesh (avocado), seeds (papaya) and kernels (mango and apricot) was carried out. The analysis of hydrocarbons by GC-MS proved the suitability of using C17:1, C16:2, C15:0 and C14:1 as markers for avocados irradiated with a low dose (0.75 kGy). The same indicators appeared following the analysis of papayas and mangoes irradiated with 1.5, and 3.0 kGy. Also, C15:0, C14:1 and C16:3 can be used to identify apricots irradiated with a low dose (0.5 kGy). The detection of alkenes was only improved by a more selective isolation, e.g. of dienes or trienes by LC-LC-GC-FID. Within a few minutes, apricots and avocados irradiated at low doses (0.5 and 0.75 kGy) can be recognized by the indicators C16:2, C17:2 and C16:3, without interfering peaks. In all cases, C16:1, C16:2, C16:3 as well as significant amounts of C17:2 can be used as markers for fruit irradiation. PMID- 8711952 TI - Determination of polycyclic aromatic hydrocarbons in smoked fishery products from different smoking kilns. AB - The levels of 13 polycyclic aromatic hydrocarbons (PAH) were determined by an HPLC method with fluorescence and UV detection and by a GC/MS technique in smoked fishery products from modern smoking kilns with external smoke generation and from traditional smoking kilns. The average benzo(a)pyrene (BaP) concentration of all 35 samples from commercial smoking kilns with external smoke generation was 0.1 microgram/kg (wet weight) and the sum of the carcinogenic compounds determined in the study, i.e. benz(a)anthracene, chrysene, benzo(b)fluoranthene, benzo(k)fluoranthene, benzo(a)pyrene, dibenz(a,h)anthracene and indeno(1,2,3 c,d)pyrene did not exceed 4.5 micrograms/kg (wet weight). The BaP levels of the 27 smoked fish samples from traditional kilns ranged from 0.2 microgram/kg to 4.1 micrograms/kg, with a mean value of 1.2 micrograms BaP/kg. The average concentration of the sum of the carcinogenic compounds was 9.0 micrograms/kg. Large variations were found in the content of the non-carcinogenic PAHs phenanthrene, anthracene, fluoranthene and pyrene in all samples from both types of smoking kilns. PMID- 8711953 TI - Influence of fungi associated with bananas on nutritional content during storage. AB - Botryodiplodia theobromae, Rhizopus oryzae, Aspergillus niger, A. flavus and Fusarium equiseti were found to be associated with the ripening of bananas and also caused rot during storage. Bananas stored in baskets with ash fire wood ripened 2-3 days earlier than bananas stored in fibre sacks and under constant light. The infected bananas showed a decrease in the quantity of total soluble sugars, protein, lipid, crude fibre, ash, ascorbic acid and mineral elements when compared with the control fruit. Paper chromatographic studies showed the presence of glucose, sucrose, fructose, maltose and raffinose in healthy control fruit, while only sucrose appeared during storage in bananas infected with B. theobromae. The total soluble sugar and crude protein contents increased during ripening. PMID- 8711954 TI - [Changes in chemical composition of tomatoes during processing]. AB - The influences of processing conditions and composition on chemical changes during thermal processing of tomatoes were investigated. During the Cold and Hot Break process pectin is degraded to different extents. Due to the thermal impact at first cyclization of glutamine to pyroglutamic acid takes place. At higher dry weight contents and during drying processes Amadori compounds and browning can be detected. The reaction products formed during thermal processing can be analysed by amino acid analysis and-after proper derivatization-by capillary gas chromatography. PMID- 8711955 TI - [Psychological stress and neuronal plasticity. An expanded model of the stress reaction process as the basis for understand central nervous system adaptation processes]. AB - A short survey on results and theories in psychosomatic and neurobiologic stress research is presented. Based on a comprehensive definition of the stress-reaction process, the biological and psychological consequences are described which are elicited by either controllable or uncontrollable stress. We conclude that controllable stress triggers the stabilization and facilitation of neuronal networks involved in the generation of appropriate patterns of appraisal and coping, whereas uncontrollable stress favours the extinction of inappropriate patterns and the reorganisation of neuronal connections underlying certain inappropriate behaviors. Both, controllable an uncontrollable stress-reaction processes are therefore essential prerequisites of, and inherent challenges to, the development and adaptation of an individual in an ever changing external world but may also lead to psychodevelopmental failures and psychosomatic diseases. PMID- 8711956 TI - [The psychotherapist and "his own" psychotherapy. Reflections on a difficult profession]. AB - In the field of psychotherapy one can see that many psychotherapists develop a special identification with the form of therapy which they learned and practice. One cannot find this kind of identification with the chosen therapeutic method in other fields of medicine (surgery, radiology, internal medicine, etc). The factors which lead to such a special identification of the therapist with the psychotherapeutic method he practices are reflected and described: They include first of all the hardly thought about motives of the therapist to chose psychotherapeutic training in a particular school. Then the influences of the peculiarities of the institutionalized work-parallel continuing education develop: The risk of emotionalized trainee-trainer dependencies, trainee-trainer identification with the associated strong delimitation towards the outside evolves. PMID- 8711957 TI - [A patient switches wards: significance for the therapeutic process in inpatient psychotherapy]. AB - The therapeutic process is influenced significantly from the outside conditions of the therapy setting. Changes in the setting in an ongoing therapy process are a special aspect. The Mannheim psychosomatic clinic has four conceptually different wards. This structure makes it possible to change settings during current therapy by moving patients from one ward to another within the clinic. With three case examples the significance of the move for the therapeutic process is described. The possibility of trial actions and gains in insight is compared with the danger of fixating pathological conflict solutions. Special attention is paid to aspects of power-powerlessness and coping with separation wishes and separation anxieties. Moving a patient to a different ward during an ongoing therapy process contains in a special way the possibility for development but also the danger of disintegration. PMID- 8711959 TI - [Psychodynamic and coping processes in patients with physical illnesses]. AB - Psychodynamic and coping processes in patients with somatic diseases are described on the background of clinical case examples. Here the connection between coping and defending is discussed whereby the special characteristics of somatic disease are taken into consideration. The secondary function, which a somatic disease can gain in an intrapsychic balance system which was already in a weak or dysregulated state is also discussed. Finally the role of social support in coping with somatic diseases is described. From this different therapeutic consequences for patients with somatic diseases are derived. PMID- 8711958 TI - [Psychosomatic determinants of restenosis after percutaneous transluminal coronary angioplasty]. AB - Coronary Angioplasty (PTCA) has become one of the standard procedures in the therapy of coronary heart disease. One of the main issues is the 20-40% incidence of restenosis of the dilated vessel. Up to now, the subject of predictors of restenosis has not yet been sufficiently clarified. Our study is concerned with two questions; first, is it possible to prove any connections between the amount of restenosis and psychosocial factors? Second, is it possible to distinguish groups of patients with and without subsequent restenosis on the basis of psychosocial variables, even before the diagnostic coronary angiography? 138 (91m, 47f) patients were assessed before the diagnostic coronarangiography, the 25 PTCA-patients (20m, 5f) were furthermore reassessed three months after the PTCA (the time of the angiographic control). Somatic and social data were collected and a semistandardised interview was conducted. In addition, coping, stress-coping control over disease and health and life-contentment were determined. Highly significant correlations was found between the amount of restenosis and resignative stress-coping, self-pity, depressive coping and flight tendency. With the help of stress-coping-subtests (which were given at the first examination) it was possible to separate the group of patients with restenosis from the group without it. PMID- 8711960 TI - [Contemporary thoughts of a psychoanalyst. On some hidden consequences of "coping with the past" in relation to the national socialism era]. AB - Many incidents awake the old suspicions about what kind of people the Germans really are, but the media are always poised to pounce on anything which looks like a reemergence of Nazi attitudes. So from this angle Germans seem to be a society which as a whole disapproves of violence and destructive behaviour. Both these images prove to be misleading. A kind of collective reaction formation has developed in Germany: intellectuals automatically assume that one can only speak about the Germans as a whole in critical and derogatory terms. Their views have developed a collective identity in which the Nazi past and the Holocaust play a central role. This deep-rooted distrust against one's own people reveals a more or less well concealed hatred directed against oneself. In the book of Alexander and Margarete Mitscherlich "The Inability to Mourn", which became a kind of credo for a whole generation of thoughtful young Germans, the authors prescribed a diet of restricted perceptions and collective self-hatred. The result is: German moral thinking shows signs of a aggressive, negative ego-ideal, and one aspect of this is collective self-castigation in the sense of a latent melancholic mechanism punishing the internalised father, because the bad past seems to have been his fault. PMID- 8711961 TI - [The potentials of computed and magnetic resonance tomography in studying peri tumorous edema and intracerebral tumors located in the supra-tentorium]. PMID- 8711963 TI - [Digital subtraction angiography and duplex scanning in the multimodal diagnosis of carotid artery pathology]. PMID- 8711962 TI - [Magnetic resonance tomography in the diagnosis of space-occupying formations in the posterior cranial fossa]. PMID- 8711964 TI - [Computed tomographic ventriculo-cisternography in assessing cerebrospinal fluid circulation in neurological diseases]. PMID- 8711965 TI - [Positron-emission tomography in the diagnosis of space-occupying formations of the brain]. PMID- 8711966 TI - [Hemodynamic and metabolic manifestations of the intracerebral steal phenomenon in arteriovenous malformations based on positron-emission tomographic data]. PMID- 8711967 TI - [Neuroradiology--its status and outlook]. PMID- 8711968 TI - [Magnetic resonance tomography in spinal cord tumors]. PMID- 8711969 TI - [The life and works of Nikolai Iakovlevich Vasin]. PMID- 8711970 TI - [Modern approaches to the diagnosis and treatment of craniocerebral trauma and its sequelae]. PMID- 8711971 TI - [The problem of treating severe pain syndromes of different origins and the contribution of Prof. N. Ia. Vasin to its study]. PMID- 8711972 TI - [The structure, goals and tasks of a future Association of Neurosurgeons of the Russian Federation]. PMID- 8711973 TI - [The organization of neurosurgical services for the population of the Russian Federation and tasks for its further improvement]. PMID- 8711974 TI - [A system of inflow-outflow drainage for neurosurgical wounds]. PMID- 8711975 TI - [Brain malformations and hydrocephalus in children]. PMID- 8711976 TI - [Magnetic resonance angiography in the neurosurgical clinic: its potentials and limits]. PMID- 8711977 TI - [Vicissitudes of treating leprosy handicaps in Kapolowe, Zaire]. AB - In 1990 Kapolowe was, without a doubt, the site of the only surgical centre in Zaire dealing with handicaps which developed in as an after-effect of leprosy. It would be useful to explain the hazards involved in such a venture for reasons which do not pertain to medicine but, rather, to particularly trying socio political circumstances. The best surgical expertise was thrown out for political reasons. Insecurity and economic hardships practically halted movement and, consequently, the wider application of such expertise. During a mission in 1994, there was a partial resumption of activities. The surgical team was reinstalled and made operational. It had been possible to state that multidrug therapy (MDT) had always ensured that the disabled leprosy patients, living in groups, and treated before 1990 under regular supervision, did not experience serious relapses. That fact corroborates earlier information relating particularly to surgical decompression. Although most of them were able to resume a certain measure of professional activity, social factors must still be borne in mind and the concept of partial permanent disability must be applied. PMID- 8711978 TI - [Results obtained by a mobile handicap-prevention unit at the Institut de Leprologie de Dakar]. AB - Of 584 leprosy patients known at the Institut de Leprologie Appliquee de Dakar because they suffered a nerve lesion with or without chronic plantar ulcer (CPU), 242 (41%) could be followed-up during a mean period of time of 8.2 years (range: 5 and 10 years) by the means of the mobile disability prevention team (health education, medical care and shoe workshop). Every two months a visit of the patients, at their home town, was organized, with the purpose to assess whether they could actually put into practice the foot and hand as having been trained for. At the same time, further advice and encouragement were given to the patients. Adapted footwear was brought to the patient, at reduced fee, the foot prints and special moulds having been taken during the previous visit. The local health worker were responsible for light surgical cares. Among the 242 followed up patients: of 107 without CPU at beginning, 90 (84%) remained so, of 135 with CPU at beginning, 57 (42%) were cured, of 135 with CPU at beginning, 74 (55%) remained stable (no worsening), the last 21, of whom 17 showed severe foot deformities but without CPU, worsened (all presented one or more CPU at the last control). Of the 242 patients, 221 (91%) remained stable or showed substantial improvement. Therefore, it must be emphasized that careful follow-up of patients is essential to insure the improvement or care of CPU as well as to prevent the onset, worsening or reappearance of CPU. Such follow-up must consist of cares, health education and special shoe wearing. PMID- 8711979 TI - Neuritic leprosy: further progression and significance. AB - Sixteen neuritic cases have been seen developing cutaneous lesions. These cutaneous lesions by and large appear within 4 months after the diagnosis of neuritic leprosy. Leprosy pathology in cutaneous lesions has been found ranging between indeterminate and borderline lepromatous group. Development of cutaneous lesions does not seem to be influenced by age, sex or number of nerves or lepromin status. Neither lesions seem to appear in any particular part of the body. Therapy, duration and type i.e. monodrug or multidrug, also does not seem to influence the development of cutaneous lesions in either way. It appears that neuritic cases with either very early (indeterminate) or with advanced multibacillary neural pathology may develop skin lesions. Skin lesion possibly appear following reversal reaction in skin. Cases with newly developed skin lesions well respond to standard therapy. Development of cutaneous lesions by neuritic cases possibly indicates towards the natural history of the disease, conforming to the hypothesis that leprosy is basically neural in inception and that all other forms emerge from it. PMID- 8711980 TI - Membrane attack complex in thickened cutaneous sensory nerves of leprosy patients. AB - Membrane attack complex (MAC) is a terminal end product produced as a result of complement activation. The deposition of MAC, in tissues, is known to have a local tissue damaging effect in several clinical conditions. Therefore, an attempt was made to demonstrate MAC in peripheral nerve biopsies, collected from leprosy patients. Interestingly, we could demonstrate deposition of MAC in involved cutaneous sensory nerves from most of the lepromatous leprosy patients. Contrary to this, majority of nerve biopsies from tuberculoid leprosy patients did not stain for MAC. Though MAC positive sections showed reactivity for S protein, our observations support the possibility that MAC, either acting directly or indirectly, may be implicated in nerve damage, at least, in lepromatous leprosy patients. PMID- 8711981 TI - Enhancement in the histological diagnosis of indeterminate leprosy by demonstration of mycobacterial antigens. AB - In a clinico-pathological study of Indeterminate leprosy, fifty-six cases were chosen based on specified clinical criteria. Their clinical features were noted, the smears for acid fast bacilli (AFB) were prepared from lesions, lepromin inoculation and biopsies were performed from the lesional edges. They were subsequently treated with a modified extended WHO regimen for paucibacillary leprosy. On routine hematoxylin eosin (HE) and Fite-Faraco staining of paraffin embedded sections, histopathological confirmation of Indeterminate leprosy was observed in only 17/56 (31%) of the clinically diagnosed cases whereas the remaining were labelled as non-specific pathology. Histometric analysis of all HE stained sections did not show any characteristic finding which could be considered as characteristic and discriminatory for Indeterminate leprosy. Immunoperoxidase staining for demonstration of mycobacterial antigen by direct staining procedure using conjugated rabbit anti-BCG and indirect three step procedure using primary rabbit anti-BCG and avidin biotin complex, was next performed on the sections exhibiting non-specific pathology. With the direct immunoperoxidase method, antigen was demonstrable in (11/35) 31% of the cases. The more sensitive indirect method could demonstrate the presence of antigen in (21/35) 60% of the cases. This study thus shows that demonstration of mycobacterial antigen by simple and inexpensive immunoperoxidase techniques enhances the histopathologic diagnosis of Indeterminate leprosy. PMID- 8711982 TI - Analysis of dystrophin in muscular diseases by two-dimensional gel electrophoresis using agarose gels in the first dimension. AB - We analyzed dystrophin in case of normal control, Duchenne muscular dystrophy (DMD), Becker muscular dystrophy (BMD) and infectious muscular disease using two dimensional gel electrophoresis and immunoblotting with 3 monoclonal dystrophin antibodies: Dys 1, a mid-rod-domain antibody; Dys 2, a C-terminal-domain antibody; and Dys 3, an N-terminal-domain antibody. In cases of normal control, a clearly separated doublet of bands was observed for Dys 1 and 3 at molecular weights 400 and 420 kDa. The isoelectric point was between pH approximately 5.7 approximately 5.9, similar to that for the myosin heavy chain. In one DMD case, a single faint band was observed for Dys 2. BMD presented a single-band pattern for each antibody. Infectious diseases cases showed 3- to 5-band patterns for Dys 1 and single or no bands for Dys 2 and 3. The pI of the Dys 1 band was almost identical. These results suggest coexistence of normal dystrophin and its proteolytic products, both containing triple helical segment, and show that two dimensional gel electrophoresis may be applicable in the analysis of dystrophin in muscular disease. PMID- 8711983 TI - The anti-Hu syndrome: a clinical and immunological study of 7 cases. AB - The authors describe the clinical and biological data of seven patients with anti Hu antibodies. Six of them displayed a small cell lung carcinoma (SCLC), but no cancer was detected in the 7th patient in spite of an extensive workup. The clinical heterogeneity of the anti-Hu syndrome is emphasized. The major symptoms were linked to a severe sensory neuropathy in three cases, to cerebellitis in two cases, to dysautonomia in one case, and to gastro-intestinal pseudo-obstruction in one case. One patient also displayed EMG abnormalities characteristic of the Lambert-Eaton myasthenic syndrome. Two patients developed opsoclonus or ocular flutter associated with severe confusion in the late stage of their disease. In four patients, the neurological signs and symptoms preceded the discovery of the SCLC, and in two cases the initial detection of anti-Hu antibodies prompted the successful search for this tumor. Immunopathological events injuring the peripheral and central nervous system are briefly discussed. PMID- 8711984 TI - [Tropical spastic paraparesis due to HTLV-I. 4 case reports and literature review]. AB - We describe the clinical, radiological, neurophysiological and biological characteristics of 4 patients suffering from tropical spastic paraparesis due to a HTLV-I infection. Patient 1 immigrated in Europe at the age of 16 from Caribbean islands. Patient 2 was a White male Caucasian who lived during several years with a prostitute also native of Caribbean islands. Patient 3 was a Black male Zairian and Patient 4 was a female of mixed race who lived many years in Zaaire and in Senegal. All the 4 patients presented with walking difficulties, a spastic paraparesis, urinary disturbances and often, severe constipation. The diagnosis was based on a positive ELISA test for HTLV-I antibodies, confirmed in 2 cases each by Western blots and detection of the viral genome by the Polymerase Chain Reaction. In all cases, CSF pleocytosis and CSF-restricted oligoclonal IgG bands were observed. MRI of the spinal cord showed a global atrophy in 3 cases. Cerebral MRI revealed unusual extensive, progressive and symmetric lesions of the white matter in Patient 1, and some patchy periventricular lesions in Patient 4; this examination was normal in the two other cases. Sensory evoked potentials indicated abnormalities of central conduction velocities at the level of the posterior columns. The pathogenesis of tropical spastic paraparesis is briefly discussed. PMID- 8711986 TI - Frontal lobe dementia presenting as personality disorder. AB - The early differential diagnosis of frontal lobe dementia is particularly difficult because changes of personality or behaviour overwhelm intellectual and cognitive impairments. We report the case of a patient presenting with paranoid personality disorder who refused to collaborate in neuropsychological testing. Initial functional imaging showed frontal hypometabolism that served as an early marker of frontal lobe dysfunction. PMID- 8711985 TI - Sodium valproate in severe migraine and tension-type headache: an open study of long-term efficacy and correlation with blood levels. AB - We performed an open prospective study of the prophylactic efficacy of sodium valproate in 56 patients among which 35 migraineurs, 7 chronic tension-type headache patients and 14 patients with combined headaches. We compared the mean number of headache days per month during a one-month-baseline period without prophylactic treatment and during the last month of a 6-month-treatment course. Among secondary parameters, we assessed headache intensity, adverse experiences and we measured valproate blood levels after one and after six months of treatment. Sixty percent of migraineurs had a 75% or more improvement in the number of headache days under sodium valproate, most of the remaining attacks being less severe. There was no significant improvement in chronic tension-type headache patients and only a mild effect in patients with combined headaches, almost exclusively on the migraine component. Thirty percent of patients reported adverse effects of which none was serious: there were 3 drop-outs. We found a moderate, but statistically significant, correlation between efficacy and blood levels of sodium valproate. PMID- 8711987 TI - Bilateral central pain. AB - A unique case of a woman with a subparietal cavernoma presenting for a time with bilateral central pain to the arms is reported. This case highlights brain mechanisms of pain processing. PMID- 8711988 TI - Idiopathic hypereosinophilic syndrome revealed by central nervous system dysfunction. AB - The idiopathic hypereosinophilic syndrome (HES) is a rare disease, characterised by persistent eosinophilia (> 1500/mm3), without underlying cause, provoking multiple organ system injury. Morbidity and mortality are mostly associated with the HES cardiopathy. Neurological signs are also frequent. Neurological dysfunction can be central (encephalopathy, organic psycho-syndrome) and peripheral (polyneuropathy, mononeuropathia multiplex, autonomic neuropathy, polymyositis). The encephalopathy is not always caused by distant thrombo-embolic events originating from the HES cardiopathy. We describe a patient with idiopathic HES central nervous system dysfunction, in the absence of cardiopathy. Furthermore we briefly discuss pathophysiological aspects, treatment modalities and the prognosis of the HES, in relation to our patient. PMID- 8711989 TI - MR features of pituitary metastases in two patients with central diabetes insipidus. PMID- 8711990 TI - The revised WHO classification of brain tumors. PMID- 8711991 TI - Traumatic brain injury, cognitive and emotional dysfunction. Impact of clinical neuropsychology research. AB - It is well established that head injury often causes brain damage leading to long term physical, cognitive and behavioural changes in the injured patients. Whereas the physical effects ranging from sensori-motor disturbances to posttraumatic epilepsy are often reported as well as cognitive sequelae, deteriorations of emotional and behavioural aspects are often neglected. Recent advances in imaging technology and clinical neuropsychology research have greatly contributed to increase our understanding of the effect of traumatic brain injury on diverse behavioural functions. After a brief review of the current status of problems facing the brain injured patients, this paper discusses the neuropsychological aspects of 3 long term brain injured patients. All 3 patients showed important behavioural and emotional distress several years after the accident. Whereas case report of patient A and C discuss the emotional and personality disturbance characterised by anxiety, depression and irritability, case report of patient C which is a case of classic frontal syndrome showed important memory impairment with emotional disturbance characterised by apathy, lack of motivation and complete indifference to his surrounding environment. Whatever the mechanisms involved, psychoaffective disturbances in the brain injured patients appear as important problems in the long term. These emotional difficulties must be considered in the treatment and rehabilitation procedures of these victims in order to help them to improve their social adjustment and quality of life aspects. Neuropsychological test data can be used to develop treatment strategies tailored for an individual's specific cognitive strengths and deficits. PMID- 8711992 TI - [Auditory brain stem evoked potentials in the evaluation of chronic fatigue syndrome]. AB - The Chronic Fatigue Syndrome (CFS) was formally defined to describe disabling fatigue of multifactorial ethology with depression and immunologic dysfunctions linked to some currently recognized infectious agents. In most cases neurophysiological tests reveal abnormalities. In this paper the Authors use low (11 pps) and high (51-71 pps) frequency ABR to evaluate the electrophysiological function of auditory brainstem responses. Eighteen patients with suspected CFS, between the ages of 17 and 63, were examined. Eleven subjects had clinically diagnosed "true" CFS (CDC criteria modified by Fukuda). The 11 pps frequency test did not reveal a high number of abnormalities in the patients in question. However, the high frequency stimulation test (with 51 and 71 pps) which was statistically significant (P = 0.009) revealed numerous aberrations in 7 patients; absence of the first wave in 1 case, in 5 numerous wave gap delays and in 1 patient absence of the first wave and numerous wave gap delays. The high frequency test did not show many abnormalities for the 4 remaining patients. For the 7 "non CFS" subjects, the clinical-audiological comparison showed no statistical significance (P = 0.920). The Authors hypothesize that the absence of the first wave in the CFS Subject may well indicate a cyto-neural junction disease in the organ of Corti. The combined analysis of clinical and audiological data showed that the described tests are more reliable when employed in dealing with patients with clinically assessed "true" CFS. PMID- 8711993 TI - [Congenital cholesteatoma of the middle ear: a case series different from cases in the literature]. AB - We present 44 cases of congenital cholesteatoma of the middle ear. Twenty-one patients had a cholesteatoma located in the posterosuperior mesotympanum. This finding was in complete contrast to the commonly reported anterosuperior location, seen in only 2 cases in our study. The remaining 21 patients had a cholesteatoma involving either the entire mesotympanum and/or epitympanum. The posteriorly located congenital cholesteatoma might represent a completely different entity and originate from epithelial cell debris trapped in the posterior mesotympanum during development of the temporal bone. All but one patient were treated with a closed tympanoplasty. Eight patients underwent single stage surgery. A preplanned second stage procedure was performed in 33 patients, while 3 are presently awaiting the second stage. Residual disease was seen in 19 patients (57%) who had undergone second stage surgery. No patient has had recurrent disease this far. Thirty-eight patients (85%) had a preoperative air bone gap of 30 dB or more. Of the 33 patients evaluated for hearing results, 16 (48%) had a postoperative gap within 10 dB. PMID- 8711994 TI - [Carcinoma of the base of the tongue: a case series]. AB - A carcinoma of the base of the tongue is diagnosed with difficulty and therapy offers a poor clinical prognosis. Between November 1988 and April 1993 45 patients were evaluated (38 men and 7 women, mean age 60 years). These patients were in poor clinical conditions and advanced clinical stage. Surgical treatment alone or in association with radiotherapy was employed in 17 patients. In the remaining patients radiation therapy alone or associated with chemotherapy was employed. Three-year overall actuarial survival was 39.5% and 3-year NED survival was 21%. Certain prognostic factors were evaluated: T stage, lymph nodes, TNM staging, extension to near anatomical areas and treatment. A Cox multivariate regression analysis revealed that clinical N stage was significant for prognosis (3-year overall actuarial survival was 76.9% for NO and 28% in subjects with clinical nodes). Subjects with neoplasm extension to pharyngo-laryngeal area could have better prognosis and, a finally, patients treated with surgery alone or in association with other treatment would achieve better local control than others. PMID- 8711995 TI - [The adipose Bichat bolla flaps in repair of oroantral fistulas]. AB - The Authors review Literature concerning surgical treatment of oro-antral fistulas. After a brief discussion concerning the anatomy of the Bichat bulla, the Authors present two clinical cases of oro-antral fistulas with purulent sinusitis. The lesions were repaired using an adipose flap of the Bichat bulla which was sandwiched between the mucous flap and the bony defect. The partial introduction of the adipose flap in to the maxillary sinus through the bony defect, enabled the osteomyelitis bony edges to revascularize and created a pathway and biological substratum for re-epithelialization of the maxillary sinus. Clinical and radiological follow-up 24 months after treatment revealed stable closure of the fistula and normal ventilation of the maxillary sinus. On the basis of these results and those reported in Literature, the Authors advise employment of this technique, particularly in extensive oro-antral fistulas which do recur with other techniques and which, furthermore are complicated by local infectious processes. PMID- 8711996 TI - [Complications of reconstructive surgery using free flaps in the maxillofacial region]. AB - The subject of our investigation was a group of twenty-nine patients with cervico facial malignant neoplasms. The 29 patients, whose ages ranged from 12 to 68, with a mean age of 40, underwent surgery between January 1992 and January 1995. Cranial base and/or maxillo-facial defects were restored by free flap tissue transfer. Thirty-one micro-surgical free flaps were performed and post-operative monitoring was carried out for about 15-20 days. The aim of the present study is to show the importance of clinical semeiology which permits an early diagnosis of post-operative complications. Furthermore, post-operative monitoring of microsurgical free tissue transfer enables an etiological diagnosis and surgical vascular salvage of the flap to be made at the most opportune time. Many Authors report microsurgical transfer success at 95%-96% because timely detection and resolution of complication can minimize flap failure. We report our experience with 31 free flaps performed in 29 patients. Three of the patients needed a second transfer due to the failure of the first. Two underwent free flap transfers, while a pedicle flap was carried out on the other. In fourteen, latissimus dorsi free flaps were used (in 2 cases the procedure had to be performed twice), the forearm and the jejunum were employed in eight patients and the rectus abdominis in one. PMID- 8711997 TI - [Extramedullary plasmocytoma (EMP) of the head and neck: a series of 22 cases]. AB - Plasmacytoma is a rare neoplastic disorder arising from B-cell series lymphocytes. It can develop in three clinical variants: a) Multiple Myeloma (M.M.); b) Solitary Plasmacytoma of the bone; c) Extramedullary Plasmacytoma (EMP). EMP generally occurs in the submucosal tissue of the upper airways (80% of cases). This paper reports 22 cases of EMP of the head and neck observed in the last 20 years and reviews pertinent Literature. Four of our cases were located in the nasopharynx, four in the oropharynx and five in naso-sinusal sites. More rarely, the lesion occurred in the larynx (2 cases) or in the oral cavity (3 cases). In 4 patients multiple localizations in the upper airways were observed. Full evaluation was carried out in order to exclude disseminated disease. With the exception of 2 cases which were surgically treated, radiotherapy (RT) represented the treatment of choice in all patients, with doses ranging from 36 to 58 Gy. In 9 cases RT was administered after complete surgical resection. Follow-up time ranged from 13 to 167 months (mean 69.6). A partial response after treatment was observed in 4 cases. In only two of these cases neoplastic residue was observed (further RT allowed complete remission of the disease). In the other two cases, residual masses consisted in amyloid deposits. Three patients developed disseminated disease (MM) after a disease-free period ranging from 3 to 6 years. Five-year actuarial disease was 92%. Although surgery is generally considered a diagnostic tool, in our opinion local disease should always be removed surgically when surgery produces low morbidity. Surgical debulking of the lesion can increase the probability or local radiotherapeutic control. PMID- 8711998 TI - [Spiral computerized tomography with tridimensional reconstruction (spiral 3D CT) in the study of maxillofacial pathology]. AB - Three dimensional computer reconstruction of CT scans provide head and neck surgeons with an exciting interactive display of clinical anatomy. The 3D CT reconstruction of complex maxillo facial anatomic parts permits a more specific preoperative analysis and surgical planning. Its delineation of disease extension aids the surgeon in developing his own mental three-dimensional image of the regional morphology. Three-dimensional CT permits a clearer perception of the extent of fracture comminution and resulting displacement of fragments. In the case of maxillo-facial tumors, 3D images provide a very clear picture of the extent of erosion involving the adjacent critical organs. Three-dimensional imaging in first generation 3D scanners did have some limitations such as long reconstruction times and inadequate resolution. Subsequent generations, in particular the spiral 3D CT, have eliminated these drawbacks. Furthermore, costs are comparable with those of other computer reconstruction technology that might provide similar images. Representative cases demonstrating the use of 3D CT in maxillofacial surgery and its benefits in planning surgery are discussed. PMID- 8711999 TI - [Endonasal dacryocystorhinostomy]. AB - Dacryocystorhinostomy an endonasal approach is a surgical procedure which permits removal of symptoms related to chronic stenosis of the naso-lacrimal duct without leaving external scars and without presenting risk for the orbital structures. The Author describes the surgical technique employed in 12 surgical cases. Use of the frontal loupe and of the operating microscope allows the surgeon's hands to remain free while all the advantages of binocular vision are maintained. Pre operative investigation must always include a dacryocystography, a CT of the paranasal sinuses and an endoscopic examination of the nasal cavities. PMID- 8712000 TI - [Primary carcinoma of the thyroglossal duct. 2 case reports]. AB - AA. present two cases of carcinoma arising on thyroglossal duct's cystic. Those carcinomas are a rare event and will be suspected in the patients that present an irregular tumefaction of medial neck's district. Etiology is unknown; is possible that a preceding irradiation on the neck will represent a factor risk. AA. retained that in case of medial neck's district tumefactions is necessary executed a total examination of neck's and head's district, an echography, a thyroid scintigraphy, a thoracic radiography, a panendoscopy and a FNAB. When we found a squamous carcinoma on residual thyroglossal duct the therapy is chirurgical in function of dimension of T and N with following radiotherapy in function of stage (possible lymph node metastasis). When we found a different adenoma is necessary chirurgical removal of tumefactions with hyoid bone's a body resection (Sistrunk's operation) with total thyroidectomy and following Pochin's test for seeking eventual residual with radiometabolic and suppressive therapy. PMID- 8712002 TI - [History corner. Pioneers of laryngology: Manuel Garcia (1805-1906)]. PMID- 8712001 TI - [Papillary carcinoma arising in thyroglossal duct cyst: a case report and review of the literature]. AB - While thyroglossal duct cysts are a frequent cause of cervical masses, carcinomas arising in thyroglossal duct remnants are rare. In this paper the Authors report a new case, which occurred in a young woman. The diagnosis of carcinomas in thyroglossal duct cysts almost always comes as a surprise upon pathologic examination. The type of neoplasia most frequently described in Literature is that of the papillary carcinoma (82%). Other types of carcinomas are also reported in Literature: mixed follicular-papillary carcinomas (9 cases), squamous cell carcinomas (10 cases) and follicular carcinomas (3 cases). There are different theories regarding the origin of and therapy for these malignancies. In the sixties some Authors thought that these carcinomas were metastases of thyroid carcinomas. Now, following demonstration of normal thyroid tissue occurrence in the wall of thyroglossal duct cysts, it is almost universally accepted that a carcinoma may arise from thyroglossal remnants. The foci of cancer found in the thyroid reported only infrequently in Literature are a result of the plurifocality of papillary carcinoma. The Authors suggest that the Sistrunk procedure is an adequate, sufficient treatment if the carcinoma is limited to the cyst's walls. They also recommend postoperative administration of substitution therapy of thyroid hormones. Indications for thyroidectomy are then discussed. PMID- 8712003 TI - [Experts' opinion on pathogenesis and classification of otitis media with effusion]. PMID- 8712004 TI - [University-hospital-territory relations in medical teaching]. PMID- 8712005 TI - [Where is Italian otorhinolaryngology going?]. PMID- 8712006 TI - [Sodium ferulate alleviates prednisolone induced liver toxicity in mice]. AB - Prednisolone (Pred) 20 mg.kg-1 sc caused an increase in serum glutamicpyruvic transaminase (sGPT) and serum glutathione S-transferase (sGST) activities and an elevation of MDA content in mouse liver homogenate. An increase of membrane fluidity of liver microsomes and mitochondria was also observed. The results indicate that the elevations of sGPT and sGST levels were related to enhancement of lipid peroxidation and membrane fluidity in Pred-treated mice. Pretreatment with sodium ferulate (SF) 100 mg.kg-1 partially alleviated the liver lesions as observed by electron microscope observation. The sGPT and sGST levels and liver homogenate MDA content were reduced, and the membrane fluidity of liver microsomes was recovered. But, the membrane fluidity of liver mitochondria was further elevated. The results demonstrate partial inhibitory effect of sodium ferulate on prednisolone-induced liver toxicity. PMID- 8712007 TI - [Effects of morphine on monosodium glutamate neurotoxicity and its mechanism]. AB - The enhancing effects of morphine on monosodium glutamate (MSG) neurotoxicity and its blocking by naloxone were studied through morphological observation, together with detection of concentrations of intracellular free Ca2+ ([Ca2+]i) by Ca2+ indicator Fura-2/AM and lactate dehydrogenase (LDH) efflux in the bathing medium in primary cultures from 14-17 d old mouse fetal cortex. It was found that 10 min pre-incubation of young cortical neurons (7 day in vitro) with morphine 10(-7) or 10(-6) mol.L-1 substantially increased LDH release from 105.7% +/- 19.0% (treated with MSG alone) to 194.5% +/- 17.7% and 214.0% +/- 9.5% respectively after exposure to MSG 0.1 mmol.L-1, but pre-incubation with morphine (10(-7) or 10(-6) mol.L-1) plus naloxone (0.1 mmol.L-1) reversed the LDH release after treatment with the same concentration of MSG. Morphine (10(-7) or 10(-6) mol.L-1) produced little elevation of [Ca2+]i. However, when combined with MSG (0.1 mmol.L-1) morphine elevated the [Ca2+]i level much more than MSG alone. These results suggest that morphine markedly enhances excitotoxic neuron damage, which can be reversed by naloxone. Overloading of intracellular Ca2+ may be a simultaneous pathological mechanism underlying the neuronal damage and death that occur in excitatory toxicity. PMID- 8712008 TI - [Effects of praeruptorin C on spontaneous contraction and action potential in cultured myocardial cells of neonatal rats]. AB - Using a circuit TV system and method of intracellular standard microelectrode, the effects of praeruptorin C on spontaneous contractile behavior and action potential were observed in cultured myocardial cells of neonatal rats. There was a decline in both the contractile frequency and velocity of cell edge motion after exposure to Pra-C for 5 minutes. Pra-C (10, 30 and 100 mumol.L-1) was shown to inhibit contraction velocity by 24%, 43% and 51%, respectively in a concentration dependent manner. Pra-C (30 and 100 mumol.L-1) inhibited the contractile frequency by 13% and 19%, respectively. Nifedipine 3 mumol.L-1 shortened APD50 and APD90 by 14% and 17% but verapamil at the same concentration inhibited APA by 27%, shortened APD50 by 8% and prolonged APD90 and SCL by 10% and 43%, respectively. Pra-C 10, 30 and 100 mumol.L-1 shortened the APD50 by 7%, 14% and 18%, respectively. Pra-C 30 mumol.L-1 inhibited APA and prolonged SCL. The results suggest that the effects of Pra-C on contractile behavior and action potential were related to its Ca2+ channel blockade. PMID- 8712009 TI - [Immunoregulatory effects of ginsenoside Rg1 in aged rats]. AB - It has been well documented that the immune function declines with age in the human and animals. The possible causes for the decline are the inability of lymphocytes to proliferate in response to mitogenic stimulation and the decrease of IL-2 production. In the present studies, Rg1 was shown to selectively enhance the proliferation of lymphocytes and the production of IL-2 in aged rats. Using Northern blot and Western blot analyses, Rg1 was found to promote IL-2 gene expression which showed increase of IL-2 mRNA and IL-2 protein contents. Interestingly, under the same conditions, studies were carried out on the effect of Rg1 on the immune function of young adult rats, but no marked influence was observed. According to these results, it is reasonable to consider Rg1 an immunoregulator rather than a purely "immunopotentiating agent". PMID- 8712010 TI - [Study on pharmacokinetics of mitoxantrone polycyanoacrylate nanoparticles freeze dried injection by HPLC column switching technique]. AB - The drug concentration--time curves of mitoxantrone polycyanoacrylate nanoparticles freeze-dried injection (DHAQ-PBCA-NP-FDIn) and mitoxantrone solution injection (DHAQ-SIn) in rabbit blood were determined by HPLC column switching technique, and the difference between the DHAQ-PBCA-NP-FDIn and DHAQ SIn in vivo was observed. The pharmacokinetic parameters of DHAQ-PBCA-NP-FDIn and DHAQ-SIn were presented by statistical moment. The results showed that the MRT and T1/2 of DHAQ-PBCA-NP-FDIn were 2.15 times longer than that of DHAQ-SIn, and the Vss was 4.81 times bigger than that of DHAQ-SIn. This indicates that DHAQ PBCA-NP-FDIn has targeting and prolonging effect on the action in vivo. PMID- 8712011 TI - [Toxicity of drugs on nasal mucocilia and the method of its evaluation]. AB - Effect of solutions or suspensions of eight drugs including analgin, paracetamol, propafenone hydrochloride, propranolol hydrochloride, ephedrine hydrochloride, gentamycin sulfate, sodium deoxycholate and hydrocortisone on ciliary movement were evaluated with in vitro or in situ toad palate model and scanning electron microscope. In vitro toad palate model: 0.2 ml of test drug solution or suspension was applied to a piece of freshly dissected upper palate of toad. The mucocilia were examined with an optical microscope and the lasting time of ciliary movement was recorded after drug application. The upper palate was rinsed with physiological saline when the ciliary movement stopped. The lasting time of ciliary movement after rinsing was then recorded again. In situ palate model: 0.5 ml of test drug solution or suspension was applied to the upper palate of toad for 30 min, and rinsed with physiological saline. The palate was dissected out and the operation was carried out in a similar manner. The results showed that the in situ toad palate model is a satisfactory method for studying the ciliotoxicity of drugs. The in vitro toad palate model is unsuitable for suspension and gel. The results of the eight drugs revealed that ciliary movement is frequently affected by many drugs and, therefore, care must be taken in developing any nasal dosage form to ensure its least ciliotoxicity. PMID- 8712012 TI - [Effect of pulse current iontophoretic transdermal delivery of insulin on blood glucose in diabetic rats]. AB - Iontophoresis of pulse current with various current intensity, frequency, on/off ratio and duration of treatment was used to facilitate the transdermal delivery of insulin in order to control blood glucose levels in diabetic rats. Male Sprague-Dawley rats were made diabetic by i.p. injection of streptozotocin (65 mg . kg-1 body weight). After two days, diabetic rats were anaesthetized with urethane (as 25% aqueous solution, 1.5 mg . g-1). Both the reservoir electrodes and the receptor electrodes were applied on the abdominal site of the diabetic rats. Iontophoresis was carried out using a prototype transdermal periodic iontophoretic system in order to provide the required direct current with desired pulse modes. The extent of reduction of blood glucose levels was found to be positively correlated with the current intensity, frequency and duration of treatment to some extent. But when the current intensity was over 0.8 mA/cm2 and the frequency was over 3000 Hz, the reduction of blood glucose levels did not continuously increase. Blood glucose levels were found to be better controlled when the on/off ratio of 1 : 1 was used. PMID- 8712013 TI - [Effects of taurine and enalapril on blood pressure, platelet aggregation and the regression of left ventricular hypertrophy in two-kidney-one-clip renovascular hypertensive rats]. AB - In two-kidney-one-clip (2k-1c) renovascular hypertensive rats, the blood pressure, left ventricular weight/body weight (LVW/BW) ratio and blood platelet aggregation were increased significantly. Enalapril (Ena) 6 mg . kg-1 . d-1 ig 9 wk and Taurine 30 mg . kg-1 . d-1 ig 9 wk can not only decrease the high blood pressure, LVW/BW ratio, but also the blood platelet aggregation induced by ADP or thrombin, though still different from that of the normal group. When the 2k-1c renovascular hypertensive rats were treated with both Ena and Tau, the blood pressure and blood platelet aggregation were decreased to the same as that of the normal group, and the LVW/BW ratio was also lowered markedly, though still higher than that of the normal group. These results show that both Ena and Tau can reverse the left ventricular hypertrophy, decrease the blood pressure and suppress the blood platelet aggregation in 2k-1c renovascular hypertensive rats. When treated with both drugs, the effects can be improved. It suggests that the two drugs can enhance the effects when used together, and that they may be two good agents for treatment of hypertension. PMID- 8712014 TI - [Studies on hepatic arterial embolization with cisplatin-chitosan-microspheres in dogs]. AB - Cisplatin-chitosan-microspheres (CDDP-DAC-MS) was chosen as a model preparation. Pharmacokinetics, targeting and embolization effects and alteration of liver function were studied after hepatic arterial embolization in dogs with CDDP-DAC MS, and the platinum content in plasma and liver was determined by flameless atomic absorption spectrophotometry (FAAS). The result of clinical trial was also reported. After the CDDP-DAC-MS and CDDP solution were respectively infused into hepatic artery of dogs through a 5F catheter, the plasma concentration of platinum and AUC of CDDP-DAC-MS group was lower than that of CDDP solution group, but the platinum content in hepatic tissue was 2.92 times as much as that when CDDP solution was given after 24 h. Angiograms revealed a remarkable decrease in the number of arterioles in liver, and histopathologic specimens showed nodular necrosis and hepatic cell degeneration in embolized region. The levels of GPT, GOT and ALP rose transiently after embolization and recovered to normal within 3 weeks. The clinical therapy in six liver cancer patients was successful. It would appear that CDDP-DAC-MS may be a good chemoembolization agent. PMID- 8712015 TI - [Protective effect of dl-3-n-butylphthalide on ischemic neurological damage and abnormal behavior in rats subjected to focal ischemia]. AB - dl-3-n-Butylphthalide (NBP) was known to have improving effect on brain energy metabolism after ischemia insult. The purpose of this study is to determine if the drug has protective action against ischemic neuronal damage. In the present study, the effect of NBP on cerebral infarction and neurological deficits after middle cerebral artery occlusion (MCAO) in rats was investigated. Focal cerebral ischemia was produced by permanent occlusion of the proximal portion of the right middle cerebral artery (MCA) according to the technique of Tamura. The infarct area was measured by 2,3,5-triphenyltetrazolium chloride (TTC) staining technique. The extent of neurological deficits was evaluated by the method of Bederson. The histological changes in neuronal change after MCAO in rats were also studied. The results indicate that the infarct area and the score of neurological deficits after MCAO were reduced significantly following intraperitoneal pretreatment or pre- and post-treatment with NBP 20 mg . kg-1. The treatment with NBP 10 or 20 mg . kg-1(i.p.), or 20,40 or 80 mg . kg-1 (po) 15 min and even 2 h (20 mg . kg-1, i.p.) after MCAO also markedly reduced the infarct area and the score of neurological deficits. However, no effect was found when NBP (20 mg . kg-1) was injected intraperitoneally 4 h after MCAO. MK-801 (0.5 mg . kg-1, i.p.), a non-competitive antagonist of NMDA receptor, significantly reduced the size of infarction and the score of neurological deficits in rats subjected to MCAO. The potency of NBP in reducing the infarct area and neurological deficits was found to be quite similar to that of MK-801 (0.5 mg . kg-1, i.p.). No neuroprotective effect of nimodipine (1.0 mg . kg-1, sc) was found. Generally, the potency of NBP in protecting rats from ischemic neurological damage is equal to that of MK-801 and is more powerful than that of Nimodipine. Side effects of NBP in behavior was not found. Therefore, NBP seems to be a hopeful drug for the treatment of stroke. PMID- 8712016 TI - [Synthesis and biological activity of dynorphin A-(1-13) analogs]. AB - Dynorphin A-(1-13) (I) and its two analogs were synthesized by solid-phase method. Fully-protected peptides were cleaved from the resin with HF. The three products were purified and their analgesic activity (antiwrithing response in mouse) and receptor affinity in mouse vas deferens (MVD) and rabbit vas deferens (RVD) were measured. The biological and pharmacological results showed that the replacement of Gla2, Ile8 and Pro10 by D-Ala8 and D-Pro10 caused an increase in analgesic activity, receptor affinity and selectivity. In analgesia and RVD assay, its activity is 2.6 times and 135 times respectively higher than dynorphin A-(1-13). The structure-activity relationships were briefly discussed. PMID- 8712017 TI - [HPLC-DAD analysis of thirteen soporific sedative drugs in human blood]. AB - An HPLC-DAD qualitative and quantitative method for the analysis of thirteen soporific sedative drugs in human blood established. These drugs were separated by HPLC with acetonitrile-water (35 : 65) (for barbital drugs), methanol-water (60 : 40) and methanol-10% triethylamine acetic acid solution (pH 7.5) (for benzodiazepam and phenothiazine drugs) as eluting phase systems and were detected with photodiode array detector (DAD). These drugs were identified by their spectral characteristics and retention times and quantitatively determined by their peak areas. In the concentration range 0.5-10 micrograms . ml-1, the concentration of all 13 drugs were in proportion to their peak areas. The correlation coefficients were all up to 0.99. The recovery rates from blood were all above 85% without interference from impurities. This method has been used to detect three poisoning samples from the clinic. Good results have been obtained. PMID- 8712018 TI - Prevalence, incidence and sex ratio of transsexualism. AB - Frequency figures for transsexualism, concerning incidence, prevalence and sex ratio calculated in the 1960s and 1970s, were compared with recent reports. The incidence figures remained constant over time, whereas the prevalence figures tended to increase during the review period. The incidence was found to be of the same magnitude in men and women, while the corresponding ratio for prevalence figures was 3:1. It is suggested that men seeking sex reassignment represent a more heterogeneous group than women, and that the reported male predominance to date is due to a lack of categorical studies of primary/ genuine transsexualism. It would appear that genuine transsexualism is, on the whole, insensitive to societal changes. The fundamental disturbance underlying this psychosexual identity disorder is suggested to be neurobiological in origin. PMID- 8712019 TI - Accuracies and inaccuracies of the family history method: a multivariate approach. AB - This paper reports on 1459 first-degree relatives of probands with schizophrenia or affective illness and matched community controls. We sought (i) to validate psychiatric diagnoses obtained by family history (FH) against those obtained by a best estimate (BE) procedure based on personal interview and (ii) to explore the factors influencing the accuracy of the FH report. We found relatively poor agreement between the FH and BE diagnoses, and the disagreements were influenced by numerous factors, including gender, psychiatric status of the informant or proband's diagnosis. When validated against a BE diagnosis, the overall accuracy of the FH method is relatively poor, and is furthermore subject to several biases. Therefore, substituting the FH method for BE diagnosis may be an important source of error in the investigation of familial/genetic factors in psychiatric disorders. PMID- 8712020 TI - Alcohol-related problems in primary care patients in Nigeria. AB - A total of 440 (50.1%) drinking patients were found in a study of 878 primary care patients in Nigeria, of whom 126 (28.6%) of drinking patients were observed to have alcohol-related problems. Those with alcohol-related problems were significantly more likely to be males, middle-aged and to belong to higher occupational groups. In addition, they were also more likely to be separated, divorced or widowed, made more frequent visits to health care facilities and were more likely to have associated mental morbidity. The primary health care (PHC) workers did not recognize these problem drinkers in their care. The need to improve the ability of PHC workers to detect and manage primary care patients with alcohol-related problems in developing countries through the use of reliable and valid short alcohol screening instruments (e.g. CAGE, AUDIT) and brief intervention techniques is emphasized. It is also suggested that, on a long-term basis, the training curricula for medical and paramedical primary care personnel in third world countries should include more hours on alcohol education. PMID- 8712021 TI - Sex differential for suicide among Austrian age cohorts. AB - The suicide mortality data in Austria were studied over a period of four decades of continuous reporting. The data were studied for age, period and cohort effects. For the period 1951 to 1990 suicides among eight birth cohorts, born between 1932 and 1975, show a marked sex differential in time trends. Up to 1985, the rates increase in a constant manner for males in all age groups, in contrast to females. Results obtained using the Poisson regression models demonstrate a 35% (incidence rate ratio (IRR), 1.35%; 95% CI, 1.27-1.45) increased risk for male cohorts born later compared with those born earlier. The risk for later-born females was not increased (IRR, 1.03; 95% CI, 0.91-1.17). Explanations such as improved living conditions for Austrian women remain tentative, however, as age, period and cohort effects cannot be separated as independent variables in the suicide mortality data available in Austria. PMID- 8712022 TI - Are all suicidal ideas closely linked to hopelessness? AB - In this study, the link between hopelessness and suicidal intent was investigated for two categories of suicidal thoughts, and the associations of these two categories of thoughts with a range of symptoms were also examined. A total of 97 patients with suicidal thoughts were assessed at the crisis unit of a psychiatric hospital. In interviews, suicidal intent was assessed using the Beck Scale for Suicide Ideation, while psychological distress was assessed using both the Beck Hopelessness Scale and the Derogatis Symptom Checklist. Ideation items describing the frequency, duration and acceptance of a wish to die were significantly correlated with feelings of hopelessness. However, items reflecting preoccupation with a method of self-harm showed only a weak correlation with hopelessness, although the relationship varied according to diagnosis. That is, this preoccupation was significantly associated with hopelessness for depressed patients, but this was not the case for the personality disorder, anxiety disorder and substance-abuse subgroups. Finally, analyses indicated that the primary predictor of suicidal intent was the patient's cognitive distortion, not hopelessness. PMID- 8712023 TI - Circadian rhythm of parasuicide in relation to violence of method and concomitant mental disorder. AB - Circadian occurrence of parasuicide was evaluated in relation to sex, violence of parasuicide method and psychiatric diagnosis. In all, 457 consecutive episodes of parasuicide were recruited during a 5-year period. Complete data for time of parasuicide, parasuicide method, parasuicide recurrency and psychiatric diagnosis (ICD-9) were available for 304 subjects. Parasuicide methods were classified into two groups depending on the violence of the method. Parasuicide occurred significantly more often in the afternoon and evening hours for both men and women, for both violent and non-violent methods, both in first-even cases and repeaters, and in the following diagnostic groups: organic mental and psychoactive substance disorders, neurotic disorders and personality disorders. The data support the hypothesis of a circadian rhythmicity of parasuicide, showing an area of chronobiological risk in the afternoon and early evening hours. PMID- 8712024 TI - Discriminant validity of the inventory to diagnose depression, lifetime version. AB - We examined the discriminant validity of the Inventory to Diagnose Depression, lifetime version (IDDL), that is, a self-report instrument designed to diagnose a lifetime history of major depressive disorder. Remitted patients with major depression (MD), remitted patients with anxiety disorders (ADs) and controls completed the IDDL. The IDDL results were then compared in the three contrasts: MD vs. ADs + controls, MD vs. ADs, and MD vs. controls. The agreement between the IDDL and clinical diagnoses was high in each of the three contrasts, and the IDDL total score in the MD group was the highest among the three groups and distinguished from the ADs group or controls. However, the IDDL occasionally diagnosed subjects with a lifetime history of ADs as having a lifetime history of MD. PMID- 8712025 TI - Incidence and sex ratio of transsexualism in Sweden. AB - The incidence and sex ratio of transsexualism in Sweden were calculated during the period between 1 July 1972 and 30 June 1992 using a case register kept on file at the Swedish Bureau of Social Welfare. Only cases of primary/genuine transsexualism were considered. The study is unique in that it has been performed in exactly the same way as a previous study by one of the authors (JW) in the 1960s and early 1970s. The data can therefore be reliably compared. The results show that the incidence figures remain constant over time and that the incidence of primary/genuine transsexualism is equally common in men and women. A larger group consisting of all those individuals who had applied for sex reassignment revealed a preponderance of men. Analysis showed that this large group included not only primary/genuine transsexuals but also effeminate homosexuals, transvestites and diagnostically uncertain cases. The importance of describing the exact methods used when calculating epidemiological data is highlighted. PMID- 8712026 TI - Parasuicide in Pakistan: experience at a university hospital. AB - There have been few reports of parasuicide from Pakistan, where the act is considered to be a criminal offence and the Islamic religion strongly disapproves of it. In order to address the problem, a retrospective case report analysis of all index cases of parasuicide presenting over a period of 3.5 years to a university hospital in Karachi, Pakistan, was undertaken. Our results showed that most of the subjects were young adults, with married women representing the single largest group. Self-poisoning with medication was the most common method, and benzodiazepines the most frequently used drug. Interpersonal conflict with the opposite sex was the most common precipitating cause. In Pakistani culture, marriage appears to be a significant source of stress for women. Reports based on official police records do not reflect the true picture of the problem in Pakistan. PMID- 8712027 TI - Psychotherapy and bulimia nervosa: evaluation and long-term follow-up of two conflict-orientated treatment conditions. AB - Analytic in-patient therapy (n = 32) and systemic out-patient therapy (n = 39) were applied to patients with bulimia nervosa, and the effects were evaluated 14, 26 and 38 months after the start of the treatments. Our assumptions about the general efficacy of both conflict-orientated techniques were confirmed: both therapies satisfactorily reduced the symptomatic behaviour, as well as secondary factors related to bulimia nervosa, in the long term. However, we could not identify differential effects of the two treatment regimes, which we had expected with regard to the very different therapeutic approaches. The similarities of outcome of both therapies predominate, with slightly better results in the case of the analytic in-patient treatment. The results are discussed with regard to the assumption that specific healing factors are involved in conflict-orientated treatments in addition to the "common factors' of psychotherapy. PMID- 8712028 TI - Charles Bonnet syndrome in leprosy; prevalence and clinical characteristics. AB - Charles Bonnet syndrome (CBS) is diagnosed when a visually impaired patient without any mental disorder develops visual hallucinations. A survey of patients in a National Leprosarium revealed that the point prevalence of CBS in leprosy was 0.4%. This prevalence appears to be high, as few cases with CBS have been reported. The semeiology of visual hallucinations was typical of CBS. However, the clinical features were different from previous reported cases because of the history of leprosy and associated multi-sensory loss. Patients with leprosy appear to be at increased risk for CBS, due to frequent eye complications combined with sensory loss, ageing, and intact intellectual functions. PMID- 8712030 TI - Psychological climate and countertransference in psychiatric treatment homes. AB - The associations between milieu therapists' feelings towards patients and the unit's psychological climate were studied at 21 small treatment homes for severely disturbed psychiatric patients. The milieu therapists filled out self report instruments where they rated their feelings towards patients on a feeling word checklist and their opinion about the psychological climate on a Bion-based scale. Scores on the climate scale were used as independent variables in a series of regression analyses, with scores on the feelings as the dependent variables. It was found that a substantial component of the average differences between units with regard to feelings towards patients could be attributed to the unit's climate. Correlations showed that units characterized by "Work' and "Pairing' had high scores for helpful and autonomous feelings, while unhelpful feelings were evoked in units characterized by "Dependency' and "Fight'. Close feelings were evoked in units characterized by "Dependency'. PMID- 8712029 TI - Inter-observer and test-retest reliability of the Italian version of the Karolinska Psychodynamic Profile in two groups of psychiatric patients. AB - A total of 64 psychiatric patients were tested with the Italian version of the Karolinska Psychodynamic Profile in order to investigate interobserver and test retest reliability. The mean inter-observer agreement coefficients (rho) ranged from 0.75 to 0.80 with less experienced psychiatrists, while the mean rho value was higher (0.96) with more experienced raters. Test-retest values were good, ranging from 0.53 to 0.93. Overall, reliability was comparable with that reported in the original Swedish study. PMID- 8712031 TI - Epidemiology of eating behaviour and weight distribution in 14- to 19-year-old Swiss students. AB - A sample consisting of 1944 Swiss students of both sexes was investigated with regard to distribution over weight categories based on BMI age percentiles and eating behaviour (EAT-26) in relation to age, sex and socio-economic status (SES). In addition, the relationship between BMI and Eating Attitudes Test scores was analysed. More subjects than expected were found to be in the obese weight categories. No clear effect of age and sex on the distribution of the sample across the five BMI classes was observed. There was an inverse relationship between BMI and socio-economic status. On the EAT scale girls scored twice as high as boys. Age had no effect on the EAT scores in females, although it did in males. Socio-economic status and EAT scores were inversely associated. Three risk categories based on the EAT scores were described: EAT 0-9 = no risk (77.5% girls, 93.0% boys), EAT 10-19 = low risk (14.1% girls, 5.5% boys), and EAT > or = 20 = high risk (8.3% girls, 1.5% boys). With regard to both aspects, weight and eating behaviour, female students showed more disturbed eating behaviour and fewer instances of deviation from normal weight. In male subjects there were more deviations from normal weight than instances of disturbed eating behaviour. PMID- 8712032 TI - An epidemiological study of eating disorders in a northern area of Japan. AB - A total of 97 patients with eating disorders who visited the Department of Neuro Psychiatry, Yamagata University Hospital, between 1978 and 1992 were studied. According to the diagnostic criteria of DSM-III-R, there were 48 cases of anorexia nervosa (AN). 38 cases of bulimia nervosa (BN) and 11 cases with both AN and BN. Both the number of patients and the ratio to all out-patients in each year increased four times during the period 1988-1992 compared to the previous period. With regard to their places of residence, the number of patients with AN and with BN from every district appeared to increase at almost the same rate, although the majority of the patients were from medium-sized cities. These findings may indicate that eating disorders are influenced by urbanization. PMID- 8712033 TI - Electrocardiographic changes in patients receiving neuroleptic medication. AB - The precise aetiology of sudden death in patients receiving neuroleptic medication is uncertain, but cardiac arrhythmias are a possible cause. We investigated the link between neuroleptic medication and electrocardiographic changes predictive of malignant cardiac arrhythmias. Electrocardiographs were performed on 111 patients receiving neuroleptic medication and on 42 unmedicated controls. Prolonged QTc intervals were more common in the patient sample, but QTc dispersion was not significantly increased. QTc interval prolongation was more likely in patients on doses above 2000 mg chlorpromazine equivalents daily (odds ratio 4.28, P < 0.02). Neuroleptic medication, especially at high doses, is associated with ECG changes that may herald more serious cardiac problems. PMID- 8712035 TI - The informational role of knowledge of results in motor learning. AB - An experiment is reported that was set-up to examine the informational role of knowledge of results (KR) in the learning of a single-limb movement timing task. A group with KR practiced 200 trials a day for 5 days prior to receiving a sixth day of practice without KR. The performance of this group was contrasted to another group that practiced 200 trials without KR for one day. Traditional movement error and time series analyses revealed that KR serves to calibrate the movement outcome to the task demands and modulate the performance outcome relation between trials. The degree of systematic trial-to-trial modulation was strongly dependent upon the degree of error exhibited on any given trial, and was enhanced under no-KR conditions. Information in KR has both immediate and persistent influences on learning and performance that are dependent upon the task constraints and the skill level of the performer. PMID- 8712034 TI - Treatment of kleptomania with fluvoxamine. AB - We report a patient fulfilling DSM-IV criteria for kleptomania who failed to respond to psychotherapy, behavioural therapy and pharmacotherapy with clomipramine, imipramine and lithium. He was successfully treated with the serotonergic agent fluvoxamine. PMID- 8712036 TI - Vergence eye movements during fixations in reading. AB - The purpose of this explorative study was to examine vergence eye movements during fixations in reading. Eye movements of twelve normal adults were assessed during reading of different materials, that is, words within context (prose passages) and words without context (word lists), as well as during different tasks, that is, reading while attending to the meaning and reading while attending to the sound (words had to be pronounced subvocally). Results indicated that vergence velocity was higher during the reading of prose than during the reading of word lists as well as higher during reading for meaning than during reading while subvocalizing. These findings were also true if only the initial 80 ms of each fixation were measured. Post-hoc analyses indicated that the effects of text type and reading objective were partially, but not entirely, attributable to differences in saccade sizes. Findings are taken to suggest that the increase in vergence velocity results from readers attending to larger units of the text. PMID- 8712037 TI - Must reasoning. AB - The mental model theory of reasoning postulates that individuals reason by constructing models of the situations described by the premises, but the number of explicit models and the information in them is kept to a minimum. The initial models of modal conditionals don't only represent the situations, but they also comment these situations, in terms of permissible and impermissible. The present paper reports three experiments which test the prediction of the mental model theory that a modus tollens inference is easier with a rule that contains the auxiliary verb 'must'. Unlike a normal rule, there is no need to flesh out the models explicitly with a rule containing 'must'. Overall, the results corroborate the prediction of the mental model theory. PMID- 8712038 TI - [Corynebacterium urealyticum or the history of an announced discovery]. PMID- 8712039 TI - [Septic lithiasis in the era of extracorporeal lithotripsy. Workshop proceedings of the LX National Congress of Urology. Santiago de Compostela, Spain, July 1995]. PMID- 8712040 TI - [Post-TUR radiotherapy in the treatment of locally advanced bladder tumor]. AB - Treatment of infiltrant vesical cancer has traditionally been the grounds for intense controversy. Transurethral resection (TUR) followed by high voltage radiotherapy (RT) in selected cases of locally advanced vesical urothelial tumours can be a valid alternative for the treatment of this condition. This paper discloses the experience of our center in the treatment of this type of tumours over the last few years (1979-1993) on 85 selected patient, with a considerable percentage of living patients and with no evidence of disease after the follow-up period; taking into account the influence on these results of the cytologic grade, tumoral size, gross appearance of the tumour, and tumoral stage. Additionally, these results are compared with those from other authors using similar therapeutic method as well as with the results from different therapeutical approaches, illustrating one more the intense controversy that exists relative to the management of these patients. PMID- 8712041 TI - [Echo-Doppler in diabetic erectile dysfunction]. AB - The erectil dysfunction is a common disorder in diabetic patients resulting from neurological, arterial and endocrine factors. This paper presents an eco-doppler evaluation of the arterial factor in 88 diabetics: 17 controlled with diet, 38 with oral anti-diabetic agents and 33 with insulin. Age, ranging from 23 to 70 years old, was similar in subjects controlled with diet (55.6 +/- 6.4) or anti diabetic agents (57.6 +/- 7.9), but greater in insulin-treated subjects (46.8 +/- 10.5) (p < 0.01). Fifty-six subjects (63.6%) has nocturnal and 43 (48.9%) diurnal erections, 45 (51.1%) achieving penetration. There was 65 (73.9%) smokers, 28 (31.8%) hypertensives, and 24 (27.3%) reported stress. A baseline evaluation was performed in all cases, in 87 after I.I.C (papaverin 26 and PgL1 61), full erection being achieved in 18 cases, non-rigid erection in 15, tumescence in 50 and with no response in 4. No differences were seen by type of diabetes. The variance analysis showed no differences in the flow speed indicators among the three groups, both at rest and after I.I.C., the sample being therefore homogeneous, superposable to arterial dysfunctions and significantly lower than those of the control group (p < 0.01). We conclude that diabetes induces arterial changes which are superposable to those caused by other vascular risk factors and independent for the type of diabetes. PMID- 8712042 TI - [PSA and PSAD study in patients with renal dysfunction]. AB - This study analyzes the changes in serum and urinary PSA values in 28 subjects; 13 with creatinine clearance under 75 ml/mn and 15 with creatinine clearance over 75 ml/mn. Both groups were compared for prostate size, measured by transrectal ultrasound, prostate weight, serum PSA (SPSA), 24h urine PSA (PSAO), PSA clearance (PSACl), serum creatinine (SCr), creatinine clearance (CrCl), PSA density (PSAD), PSA/creatinine ratio (PSA/Cr) and PSACl/CrCl ratio. Mean values of SPSA and PSAO were 4.5 +/- 0.8 and 222 +/- 29.7 ng/ml respectively, values for SCr, CrCl and PSACl averaging 1.62 +/- 0.2 mgr/dl, 71.6 +/- 6.5 ml/mn and 150.5 +/) 32.9 ml/mn. Median prostate size was 32.6 +/- 3.9 cc, with weights of 40.3 +/ 4.9 g and mean PSA density (PSAD) 0.13 +/- 0.02. The results of the homogeneity study showed that there are no significant differences between both groups with regard to the variables considered in the study. SPSA values were higher in patients with CrCl < 75; 3.4 vs 5.7, but not significantly. There are no significant differences between PSAO and PSACl values for both groups, even though PSAO levels were higher in patients with CrCl < 75 ml/min (p = 0.1). PSAD values for patients with CrCl > 75 ml/mn were lower than those for patients with CrCl < 75 ml/mn; 0.09 vs 0.17 (p = 0.08). In the entire sample, PSAD levels showed correlation with SPSA and PSA/Cr values; R = 0.63 (P = 0.0003) and r = 0.5 (p = 0.009) respectively. Also, they were significantly but inversely correlated with PSACl levels; r = - 0.5 (p = 0.006) and PSACl/CrCl; r = - 0.048 (p = 0.01). No correlation was seen between PSAD values and the following parameters; PSAO (p = 0.7), SCr (p = 0.5) and CrCl (p = 0.27). When the group of patients with CrCl < 75 ml/mn is considered, PSAD values are correlated exclusively with PSACl values; r = - 0.69 (p = 0.008) and PSACl/CrCl; r = 0.68 (p = 0.009). Our data appear to indicate that there is a certain relationship between PSAD and the renal function although the physiopathological mechanism responsible for that is unknown. Nevertheless, considering the sample size, more comprehensive studies will be necessary to obtain more convincing results. PMID- 8712043 TI - [Chronic prostatitis: diagnostic and therapeutic considerations]. AB - Chronic prostatitis, whether of bacterial or non-bacterial origin, is a pathological entity commonly considered difficult to diagnose and to manage. In an attempt to provide a better understanding of the usefulness of Stamey-Meares fractional culture as a diagnostic test, the findings obtained with this technique in 34 patients, 5 with bacterial chronic prostatitis (BCP) and 29 with non-bacterial chronic prostatitis (NBCP) were analyzed. The multiresistance found in the antibiogram in 3 cases and the lack of correlation between these findings and the seminoculture, demonstrate that this technique should continue to be considered indispensable. Also, the value that should be appointed to the existence of organisms with non-proven pathogenicity is discussed. Some bases for a rational management are proposed and the various treatment alternatives, including the use of myorelaxants which in this series allowed to achieve resolution of the symptomatology in 59% cases are analyzed. PMID- 8712044 TI - [Nosocomial infection in BPH: economic costs and increase in hospital stay]. AB - We present a cohort study where 40 matched pairs of patient (infected and no infected) were included. All of the patients had prostatic benign hyperplasia and open surgery or thransuretral resection was developed. Infected patients stay 3 days more in the hospital and they cost 120.000 pts more than uninfected patients. PMID- 8712045 TI - [A new method for assessing sphincter resistance, in patients with low compliant bladder]. AB - Since 1990, in our department we have planned out a test to find out the grade of sphincteric resistance in patients with low compliant bladders. In order to know which patients will have a sufficient sphincteric resistance to achieve satisfactory continence after bladder augmentation and which patients will not have sufficient sphincteric resistance, needing procedures to increase outlet resistance along with bladder augmentation. This test consisted in: to determine what volume the bladder can admit at a detrusor pressure of 10 cm H2 O, we fill the bladder at that volume and then, we check if the patient had leaks with abdominal pressure increases. The study was carried out in 15 patients with low compliant bladders and suffering from incontinence, who had not responded to I.C. and anticholinergic therapy, who was to undergo bladder augmentation. In 12 patients the test was negative and in 3 was positive. After bladder augmentation, all patients with negative tests, are completely dry, and the three patients whose test was positive, in spite of having urodynamically normal neobladder they remain incontinent. In our opinion, this test is simple to perform and very useful diagnostic tool to known which patients beside bladder augmentation, may need procedures to increase outlet resistance. PMID- 8712046 TI - [Bladder carcinosarcoma. A new case]. AB - Vesical carcinosarcoma is a very poor prognosis, infrequent tumour normally presenting with haematuria and for which the only treatment available is radical cystectomy. Here we present a new case of this tumour in a 61 year-old male with a brisk evolution to death. PMID- 8712047 TI - [Urologic impact of Crohn disease]. AB - Presentation of two cases of Crohn's disease seen in our Service in which the urological symptoms were of special relevance. One patient presented clinically with a picture of anaemic gross haematuria. The other one, was a patient already diagnosed with Crohn's disease who developed an enterovesical fistula in spite of receiving medical treatment. Both cases were resolved surgically. The clinical aspects, natural history and treatment of this uncommon form of urinary tract involvement are discussed. PMID- 8712048 TI - [Stasis priapism secondary to PGE1: transformation into high-flow priapism]. AB - Since vasoactive agents were first introduced for the diagnosis and treatment of Erectile Dysfunction (ED), the most feared side effect has been priapism. The arrival of Prostaglandin E1 (PGE1) limited to a large extent the incidence of this complication due to its fast systemic and intracavernous metabolism. A case of stasis priapism following a low dose PGE1 injection (10 mcg) after 48 hours evolution and its surprising conversion to high flow priapism after the surgical procedures performed to solve this complication is presented here. PMID- 8712049 TI - [Secondary hypertension of equivocal diagnosis]. AB - Secondary hypertension (20%) is the result of specific causes and is susceptible of cure. It is uncommon, however, that in the same patient several pathological conditions which a priori could individually cause increased pressure values should coexist. A presentation if made of a patient with arterial hypertension, the study of which demonstrated the existence of homolateral adrenal tumour (corticoadenoma), renal tumour (angiomyolipoma), as well as proximal stenosis and small distal aneurism of the renal artery upper branch. An explanation is given here of the diagnostic evolution showing the images obtained, including a debate of the methodology followed and treatment performed. PMID- 8712050 TI - [Laparoscopic bilateral nephrectomy of native kidney in kidney transplantation carriers]. AB - Severe vesicouretheral reflux sometimes causes terminal renal failure which then requires renal transplantation. In these patients it may be necessary to perform bilateral nephrectomy when associated to serious hypertension or urinary infections with a significant clinical impact. This paper presents the clinical picture of a patient with a corpse renal transplantation, due to terminal renal failure secondary to reflex nephropathy, in whom it was necessary to perform bilateral nephrectomy due to the uncommon urinary infections requiring hospitalization for treatment. The bilateral nephrectomy was done through transperitoneal laparoscopic approach, using seven points for trocars introduction. The procedure lasted 4 hours and the patients was discharged within 48 hours with no complications. Accumulate experience in the use of laparoscopy in urology has allowed this technique to be feasible. After designing the experimental model, our group has had the chance to accomplish laparoscopic nephrectomies in humans. We believe that this is the first case of bilateral nephrectomy conducted in our country using the laparoscopic approach. PMID- 8712051 TI - [Urodynamic picture in high spinal cord ischemia, induced by addictive use of cocaine. Report of a case]. AB - The authors present a case of spinal cord ischemia at level C2 related to cocaine use. The neurologic pattern was an anterior cord hemisection syndrome. Urodynamic assessment was performed at 6 and 14 months from the neurovascular event. The ice water test (IWT) showed an uninhibited bladder cooling reflex, despite an areflexic detrusor (at routine filling cystometry). Occult bladder hyperreflexia became overt during the following month and was registered by the second urodynamic assessment. Anatomical bases of spinal cord blood supply are discussed. The authors emphasize the role of the IWT in predicting the development of overt bladder hyperreflexia and thus, spinal shock phase resolution. PMID- 8712052 TI - [Current treatment of ureteral lithiasis]. PMID- 8712053 TI - Aggression and hostility in substance abusers: the relationship to abuse patterns, coping style, and relapse triggers. AB - A cohort of 3,367 substance abusers seeking treatment were administered measures of aggression and hostility including the Buss-Durkee Hostility Inventory and the NEO Personality Inventory Hostility Scale. Polysubstance abusers scored significantly higher on all measures of hostility and aggression, regardless of whether they abused cocaine or not. Subjects scoring higher on aggression and hostility utilized escape-avoidance, distancing, and confrontational coping styles more regularly. Subjects scoring higher on measures of aggression and hostility reported more situations that triggered their use of substances and less confidence that they could resist using when faced with such situations in the future. This was especially true for situations involving unpleasant internal states, situations involving rejection, and situations involving conflict with family and friends. The implications of these findings for clinical assessment and treatment planning are discussed. PMID- 8712054 TI - The burden of smoking-attributable mortality among African Americans--Indiana, 1990. AB - Recent trends in smoking prevalence in Indiana indicate a slow decline in smoking prevalence among adults aged 18 years of age and under. Despite this decline, the smoking prevalence has remained consistently higher among Blacks compared to Whites. The purpose of this investigation was to evaluate the extent of mortality due to cigarette smoking in 1990 among Blacks and Whites in the state of Indiana. Estimates of smoking-attributable mortality (i.e., smoking-attributable mortality [SAM] and years of potential life lost [YPLL] associated with smoking in 1990 were calculated using SAMMEC, a software program developed by the Centers for Disease Control and Prevention and designed to measure the disease impact associated with cigarette smoking. Although the SAM and YPLL for Whites accounted for the greatest percentage of total SAM and total YPLL, SAM rates and YPLL rates were higher among Blacks compared to Whites, for both men and women. The SAM rate for Black men was 4% higher than that of White men and 9% higher for Black women compared to White women. Similarly, the YPLL to life expectancy rate for Black men was 7% higher than the YPLL rate for White men and 29% higher for Black women compared to White women. The greater burden of tobacco-related mortality among Blacks is evidenced through higher SAM and YPLL rates compared to Whites. It is imperative that high-risk populations be targeted at national, state, and local levels through anti-tobacco campaigns. PMID- 8712055 TI - Autonomic cued reactivity in alcoholics: the effect of olfactory stimuli. AB - The present study was designed to investigate autonomic cued reactivity to olfactory alcohol stimuli in alcoholics. Twenty outpatient alcoholics and 20 social drinkers were exposed to high- and low-potency alcohol and neutral odors. The alcoholics showed greater skin conductance responses and increased heart rate acceleration responses to the high-potency alcohol odor than social drinkers, while there was no difference between the groups' responses to the low-potency alcohol odor. Alcoholics also reported greater difficulties in resisting a potential offer for a drink after relative to before the experiment, while there was no change in the desire to drink. The results indicate that alcohol cues are perceived as emotionally aversive and elicit a defensive response to avoid further processing of these stimuli. The increased autonomic reactivity may thus reflect a shift of focus from the environmental alcohol cues to internal thoughts and feelings. A rigid internal focus may constrain the ability to resist alcohol consumption and thus be a critical determinant in promoting craving and relapse in alcoholics. PMID- 8712056 TI - On the relationship between emotional and external eating behavior. AB - Although there is a strong relationship between emotional and external eating, separate subscales for these behaviors have been constructed in the Dutch Eating Behavior Questionnaire. This study tries to establish whether this distinction is justified. We studied relationships among self-reported ( 1) degree of emotional and external eating behavior and (2) problems with (a) emotional distress and relationships, (b) stimulus-boundness (inappropriate amounts of either too much or too little exercise, work, leisure activities, and spending money), and (c) problems with substance use (alcohol, illicit drugs, nicotine, or caffeine) in a sample of female students. No relationships were found between either type of eating behavior and problems with substance use. Furthermore, the significant relationship between emotional and external eating behavior and stimulus boundness disappeared in the subsample who had problems with overeating. The fact that in all samples emotional eating was significantly related to problems with emotional distress and relationships (anxiety, depression, phobias, suicidal acts or ideations, intimate relations, and sexual contacts) but external eating was not, suggests that the two types of eating behaviors refer to independent constructs. Thus, the use of separate scales to measure these theoretically different aspects of overeating seems warranted. PMID- 8712057 TI - DSM-III-R tobacco dependence and quitting during late adolescence. AB - Previous studies of tobacco dependence have tended not to examine adolescent samples. In a cohort of 18-year-olds, the 12-month prevalence rate of dependence based on DSM-III-R criteria was found to be 19.3%. Cotinine levels in saliva were positively related to level of cigarette consumption but not to tobacco dependence and may not be a suitable measure of dependence. Measures of behavioral and psychological aspects of smoking supported the utility of DSM-III R criteria for tobacco dependence among young smokers. As a large number of young smokers are dependent on tobacco. the results support the development of treatment programmes for youth that take into consideration their degree of dependence. PMID- 8712058 TI - The extinction of naturally occurring conditioned reactions in psychoactive substance users: analog studies. AB - In the present series of studies we develop an analog approach for the study of conditioned reactions to drug stimuli. The analog we study is the naturally occurring conditioned reaction of salivation at the sight of a lemon. We show that this conditioned reaction can be extinguished, that spontaneous recovery occurs, and that the conditioned reaction increases after "relapse." Further, we show that massed extinction trials lead to greater extinction than do spaced trials. This analog provides an approach that can be used to develop cue-exposure treatments that minimize spontaneous recovery from extinction and reduce the likelihood of relapse. PMID- 8712059 TI - Parental alcohol abuse and gender as predictors of psychopathology in adult children of alcoholics. AB - By means of procedures designed to address methodological weaknesses of previous research, this study tested hypotheses regarding the presumed greater tendency of adult children of alcoholics (ACA) to be characterized by symptomatology indicative of psychopathology than adult children of nonalcoholics (ACNA). Gender of ACA and gender of alcoholic parent were also examined as predictors of psychopathology in ACA. Four-hundred-twenty-five undergraduate students completed the Clinical Analysis Questionnaire to assess anxiety, depression, and general maladjustment, and the Children of Alcoholics Screening Test to identify ACA (88 females, 30 males) and ACNA (235 females, 72 males). ACA reported more symptomatology indicative of depression and general maladjustment than did ACNA. but gender of ACA and gender of alcoholic parent were not significant predictors of psychopathology among ACA. PMID- 8712060 TI - Transitions to cigarette smoking during adolescence. AB - The process of transition from nonsmoking to regular weekly smoking during a period from 10 to 16 years was examined using data gathered during the course of a longitudinal study of 957 New Zealand adolescents. These data were analyzed using a latent Markov model to estimate both rates of transition between stages of smoking and errors of measurement in report data. The fitted model suggested that the process of transition to smoking was a progressive and one-way process in which once teenagers had graduated to a given smoking status, return to earlier stages was uncommon. This process also showed a clear tendency to accelerate with age, so that rates of transition to smoking behaviors after the age of 14 years were far higher than rates of transition at age 10 years. The model also made it possible to estimate the accuracy of smoking reports. This suggested that individuals who were nonsmokers or regular smokers were classified with better than 95% accuracy on the basis of report data. However, the reporting accuracy of occasional smoking was poor, with 42% of occasional smokers being falsely classified as nonsmokers. The implications of these findings for the understanding of the processes of transition to smoking behaviors and the effects of measurement errors in report data are discussed. PMID- 8712061 TI - Understanding comorbidity between PTSD and substance use disorders: two preliminary investigations. AB - While there is high level of comorbidity of PTSD and substance use disorders (SUDs), little research has focused on the overlapping symptom constellation characteristic of both PTSD and substance use/withdrawal. This report describes two preliminary investigations that address this area. In the first study, the pattern of PTSD symptoms in a sample of women (n = 28) seeking treatment for a SUD and comorbid with PTSD was compared with the symptom pattern of a sample of women (n = 28) with PTSD only. The PTSD + SUD group evidenced significantly more symptoms in the avoidance and arousal symptom clusters than the PTSD-only group. At the individual symptom level, the PTSD + SUD group reported significantly more sleep disturbance than the PTSD-only group. It was also determined that the PTSD + SUD group reported greater traumatic-event exposure than the PTSD-only group. In the second study, PTSD symptoms were compared in a sample of alcohol- dependent and a sample of cocaine-dependent individuals with PTSD. The alcohol- dependent group exhibited significantly more arousal symptoms than the cocaine dependent group. Implications of the results for the assessment of individuals with comorbid PTSD and SUDs are discussed. PMID- 8712062 TI - Effect of smoking cues and cigarette availability on craving and smoking behavior. AB - This study examined whether smokers respond differently to smoking cues than to affectively neutral or unpleasant cues without smoking content, and whether reactivity is affected by expectations regarding the opportunity to smoke. Expectancy was manipulated by telling subjects in group SMOKE that they could smoke, and subjects in group NO SMOKE that they could not smoke following each cue-reactivity session. The dependent variables were subjective ratings of "desire to smoke", "high", "withdrawal", and mood, as well as latency to initiate smoking measured in group SMOKE. Statistical analyses demonstrated that only group SMOKE (a) reported greater "desire to smoke" and "withdrawal" to the smoking cues compared to the baseline, (b) reported greater "desire to smoke" to the smoking cues than to the unpleasant or to the neutral cues, and (c) smoked faster after the smoking cues than after the neutral cues. Both groups rated the unpleasant cues as affectively more negative than the neutral cues. The data suggest that the impact of drug-cues on craving and subsequent drug-use is due to their drug-related content, and not to negative affect. Furthermore, the impact of drug-related cues appears to be influenced by perceived drug-availability. PMID- 8712063 TI - Characteristics of youth and young adults seeking residential treatment for substance use problems: an exploratory study. AB - In a convenience sample of young people (aged 12-25) seeking help for substance use, those preferring residential treatment had more mental health and addiction problems than those preferring nonresidential treatment. Those seeking residential treatment were also more likely to have had previous treatment experiences. Some clinical and research implications are noted. PMID- 8712064 TI - Judgment of intoxication by untrained social drinkers across drinking settings. AB - Subjects' perceptions of cues used to judge intoxication across different drinking setting were investigated. Drinkers at a fraternity dance party and in a college town bar rated the importance of 18 cues for making estimations of their level of intoxication. Results indicated that 11 of the 18 cues were rated as more important in the party situation than in the bar situation. These results suggest that subjects apply cues to intoxication differently in different situations. PMID- 8712065 TI - Psychiatric symptoms in polysubstance abusers: relationship to race, sex, and age. AB - The Symptom Check List 90R (SCL-90R) was administered to a sample of 94 polysubstance abusers seeking outpatient detoxification for opioid addiction. The relationships among race, sex, and age, and psychiatric symptoms were examined. Results obtained by an ANOVA indicated a significant effect only for race. White substance abusers reported significantly more psychiatric symptoms (on 9 out of 12 SCL-90R scales) than did Black substance abusers. A comparison of effect sizes between those found in the present study and those of related studies is also presented. Results suggest that race differences may be relevant to treatment resource and treatment planning. PMID- 8712066 TI - Viruses in human tumors--reminiscences and perspectives. PMID- 8712067 TI - Proteins of the Myc network: essential regulators of cell growth and differentiation. PMID- 8712068 TI - The role of the insulin-like growth factor system in human cancer. PMID- 8712069 TI - Genetic basis and clonal evolution of human prostate cancer. PMID- 8712070 TI - Signaling by the cytokine receptor superfamily in normal and transformed hematopoietic cells. PMID- 8712071 TI - Genetic alterations of cyclins, cyclin-dependent kinases, and Cdk inhibitors in human cancer. PMID- 8712072 TI - [Trends of research in pathophysiology of optic nerve disorders]. PMID- 8712073 TI - [Ophthalmopathy due to environmental toxic substances especially intoxication by organophosphorus pesticides]. AB - Organophosphorus pesticides (OPs) produce optico-autonomic peripheral neuropathy in human populations in districts where a large amount of pesticides have been used in agriculture. This report presents an epidemiological study that was performed in Kanagawa Prefecture. An autopsy case of a professional organophosphorus sprayer is reported. In addition, an experiment was performed to investigate a non-cholinergic chronic toxicity due to a certain OP. The epidemiological study revealed that 64 of 7,435 farmers showed vertical smooth pursuit defect of the eyes, impairments of modulation-transfer function (MTF) of the visual system and abnormal contraction dynamics of the pupil reaction to light stimuli, and a high residual level of OP was found in their blood. These abnormalities were reduced by treatment with antidotes such as atropine, prifinium bromide, and pralidoxime methiodide (PAM). The autopsy findings showed severe retinal degeneration with optic neuropathy and an obviously precocious progression of arteriosclerotic change in heart, brain, and retinal vessels. These findings can not be explained by cholinesterase inhibition alone. Experimental evidence showed that OP produced non-cholinergic impairment such as increase of Ca ions in retinal neurons. A generation of free radicals was noted in tissue-cultured retinal neurons. Blood selenium level was reduced by OP. These non-cholinergic actions may also explain the neural damage caused by long standing low-dosage contact with OPs. PMID- 8712074 TI - [Distribution of conjunctival goblet cells and observation of goblet cells after conjunctival autotransplantation in rabbits]. AB - Conjunctival goblet cells are essential for maintaining a normal ocular surface and important for recovering a diseased ocular surface. However, how goblet cells in conjunctival epithelium evolve after transplantation are uncertain. We experimented with albino rabbits to study the distribution of goblet cells in normal conjunctiva. We performed autograftings of conjunctiva which contained many goblet cells on the sclera of albino rabbits, and made histological observations on how goblet cells evolved after transplantation. The results were as follows. 1. Goblet cells in the normal rabbits were most abundant at the lid margin of both upper and lower tarsal conjunctiva. 2. We made autograftings of conjunctiva on the sclera of albino rabbits. The goblet cells on the transplanted conjunction disappeared for a short time on the 3rd day after transplantation. On the 7th day and onwards they reappeared, and by the 14th day a large number of goblet cells were observed. 3. Between the 7th and 14th day after autografting, epithelial differentiation of the grafted conjunctiva occurred. PMID- 8712075 TI - [Comparative measurement of optic nerve head circulation using laser speckle circulation analyser of ocular fundus and a hydrogen clearance method]. AB - The microcirculation in the optic nerve head (ONH) was measured using the recently developed laser speckle circulation analyser for the ocular fundus. The measurements were compared to those of the capillary blood flow (CBF) obtained by the hydrogen clearance method. The ONH blood flow was measured using both methods simultaneously in six normal albino rabbits. The blood flow was increased by inhalation of carbon dioxide (CO2) and reduced by systemic administration of endothelin-1 (ET-1). 10%CO2 inhalation showed a CBF increase of about 22% in the ONH in the hydrogen clearance method, and intravenous injection of ET-1 at 10( 10) mol/kg decreased the reading by about 19%. A significant correlation (r = 0.92, p < 0.01) was found between the relative values of NB (Normalized blur, by laser speckle method) and CBF in the ONH. This result indicates that the change of NB in the ONH shares the same accuracy as hydrogen clearance method within the range of our present experiment. This study provides a fundamental basis for the clinical application of the laser speckle method in measuring ONH circulation. PMID- 8712076 TI - [The effect of hyaluronic acid on keratocyte proliferation]. AB - Our previous study demonstrated that during healing of penetrating corneal incision, hyaluronic acid (HA) appears at the wound area with massive infiltration of activated keratocytes, suggesting a possible role of HA in keratocyte proliferation. In the present study, we investigated the effect of HA on keratocyte proliferation. A penetrating incision was made in the center of rabbit corneas. Wounded corneas were treated with HA eye drops or physiological saline every 2 hours for 24 hours. Then the corneas were excised, labeled with 3H thymidine for 4 hours, and subjected to autoradiography. Rabbit keratocytes were cultured in chamber slides in TC199 medium alone or in TC199 containing HA for 24 hours, labeled with 3H-thymidine for 4 hours, and subjected to autoradiography. In the corneas treated with HA, the number of keratocytes incorporating 3H thymidine was significantly higher than in the control corneas. The rate of 3H thymidine uptake in keratocytes cultured with any concentration of HA was higher than that of the control. Both in vivo and in vitro studies demonstrated stimulatory effect of HA on keratocyte proliferation. It is possible that HA might promote corneal stromal wound healing by stimulating keratocyte proliferation. PMID- 8712077 TI - [Ultrastructural study on rat precorneal tear film by quick freezing-freeze substitution method]. AB - The ultrastructure of rat precorneal tear film was visualized by the quick freezing-freeze substitution (QF-FS) method and the conventional fixative method under the scanning electron microscope (SEM). In the QF-FS method, the eyeballs of rats were quickly frozen with an isopentane-propane mixture cooled by liquid nitrogen applied directly to the eyes. After enucleating the eyes and fracturing the frozen corneas, the corneas were prepared for SEM observation. In contrast to the conventional fixative method, by which the microvilli of the surface epithelial cells could be visualized clearly, a very thin membrane-like structure was observed to cover the corneal surface in the eyes prepared by the QF-Fs method. Between the membrane-like structure and corneal surface, a homogeneous and fine network-like structure was observed. The results suggest that the structure of the tear film might be different from the one we have believed to consist of three layers until now. The QF-FS method is considered to be useful for the morphological study of the precorneal tear film. PMID- 8712078 TI - [Vitrectomy with autoserum for idiopathic macular hole]. AB - Using autoserum, we operated on eyes with full thickness macular hole and achieved greater anatomic and visual success than with eyes that underwent vitrectomy alone. A total of 34 eyes were treated with vitrectomy and fluid-gas exchange: 14 eyes with autoserum instillation and 15 eyes without autoserum. Five eyes with macular holes persisting after initial vitrectomy underwent reoperation with autoserum. After vitrectomy and fluid-air exchange, 0.1 to 0.3 ml of autoserum was placed for about 7 minutes over the hole, and then 30% SF6 was instilled. After treatment, successful anatomic closure occurred in 53% of the eyes without autoserum, in 86% of those with autoserum, and in 80% of those which underwent reoperation with autoserum. Visual acuity improved 2 lines or more in 53% of the eyes without autoserum, in 79% of those with autoserum, and in 60% of those that underwent reoperation with autoserum. The results of the current preliminary study shows that treatment of full-thickness macular holes with autoserum seems to be more beneficial than vitrectomy alone. PMID- 8712079 TI - [A case report of Mycobacterium chelonei keratitis]. AB - We treated a patient with keratitis caused by Mycobacterium chelonei (M. chelonei). The patient was a 74-year-old man who had been successfully treated for stromal type herpetic keratitis with topical steroids. The corneal lesions appeared chalky white with indistinct margins and had radial projections in deep level stroma. M. chelonei was found in second corneal scrapings. Topical tobramycin that had sensitivity resolved the keratitis rapidly. M. chelonei keratitis has been reported in only 25 cases so far and this case is the first report in Japan. M. chelonei keratitis occurred mostly in eyes which had been subjected to trauma, surgery, and the use of topical steroids. If these characteristic corneal findings appear in such predisposed eyes, M. chelonei keratitis should be considered. PMID- 8712080 TI - [Histochemical studies on two cases of Peters' anomaly]. AB - We examined glycosaminoglycan molecular species in the corneal stroma and the sclera histochemically in two cases of Peters' anomaly. Paraffin-embedded sections were stained with either hematoxylin-eosin or sensitized high iron diamine combined with enzyme digestion and then examined with a light microscope. In the center of the cornea, the histological specimens revealed defects of the corneal endothelium and Descemet's membrane, together with a local defect of the corneal stroma. Disorganized lens materials were detected in the corneal stroma. In the peripheral cornea, the undifferentiated iris adhered to the corneal posterior surface. Based upon the findings mentioned above, these two cases were diagnosed as having Peters' anomaly caused by faulty separation of the lens vesicle. The sclera showed normal structures in the specimen stained with hematoxylin-eosin. In both cases, the corneal stroma involved nearly equal amounts of chondroitin sulfate A/C and B. The sclera, on the other hand, contained a large amount of chondroitin sulfate A/C and a relatively small amount of chondroitin sulfate B. Keratan sulfate was never detected in either the cornea or the sclera. Thus, disorders of glycosaminoglycan molecular species were detected in the sclera as well as in the corneal stroma. PMID- 8712081 TI - [Two cases of systemic lupus erythematosus presenting with disc edema]. AB - We report two young women, 22 and 19 years old, who showed bilateral optic disc edema in the course of systemic lupus erythematosus. Lumbar puncture showed increased intracranial hypertension with no abnormal findings in the composition of the cerebrospinal fluid. Computed tomography and magnetic resonance imaging showed no abnormal findings. They were diagnosed as having rare intracranial hypertension associated with systemic lupus erythematosus. Treatment with systemic corticosteroids produced a dramatic resolution of the increased intracranial hypertension and the disc edema. PMID- 8712082 TI - [Study of twenty-three cases of pediatric testicular tumors]. AB - Twenty-three cases of testicular tumor (10; malignant, 13; benign) in childhood are reported. The age at diagnosis was between 3 months old and 8 years old. Twelve patients visited the clinic within a month of first appearance of symptoms. All but one patient received high orchiectomy. One patient had resection of tumor because of its benign property. Eight of the 10 patients with malignancies received 30-50Gy of retroperitoneal radiation, including one patient (stage IIa) with retroperitoneal lymph node dissection. Two patients (stage I and IIa) with post-operative metastasis in lungs, brain and mediastinal lymph nodes were treated with chemotherapy repeatedly, but they died. Our latest therapy is; 1) high orchiectomy recommended for benign and stage I malignant testicular tumors in childhood, 2) retroperitoneal radiation and/or lymph node dissection limited for patients with stage II malignancy, and 3) Systemic chemotherapy for stage III malignant tumors. At least three years of follow up would be needed. Those who received radiation or chemotherapy should be followed longer. PMID- 8712083 TI - [Long-term effect after total parathyroidectomy with autotransplantation for secondary hyperparathyroidism]. AB - Although the short-term outcome of total parathyroidectomy combined with autotransplantation in cases of renal osteodystrophy has been reported by many investigators, few studies have been made on the long-term outcome of this surgical technique. We recently examined the long-term outcome of this surgery by following 19 cases for more than one year (range: 12-70 months, mean: 31.7 months). During the follow-up period, changes in subjective symptoms, biochemical parameters and bone mineral density (BMD) were monitored. At the end of the follow-up period, C-parathyroid hormone (C-PTH) and alkaliphosphatase (ALP) were significantly lower than their preoperative levels. Ca, P and %BMD showed no significant change from their preoperative levels, although %BMD tended to be higher than its preoperative level. Depending on the presence or absence of osteoarticular pain at the end of the follow-up period, the patients were divided into the improved group and the non-improved group. Of the 5 patients allocated to the non-improved group, 4 were female and only 1 was male. C-PTH and ALP were significantly higher in the non-improved group. %BMD was higher in the improved group than in the non-improved group, although this difference was not significant. In the non-improved group, 2 patients were suspected of having extra parathyroids, 1 was suspected of having recurrence, and 2 were suspected of having postoperative osteomalacia. Postoperative reduction in BMD was only seen in females, suggesting its relationship to postmenopausal osteoporosis. PMID- 8712084 TI - Effects of Ischemia on the function of the isolated rat detrusor muscle. AB - Ischemia of vital organs causes various degrees of impairment. We studied the in vitro effects of ischemia on the function of the rat bladder. Ischemia was induced by ligation of the bilateral (bilateral ischemia) or right (unilateral ischemia) internal iliac arteries. Bladder weight increased significantly following 1 week of bilateral and unilateral ischemia. Passive tension was significantly higher in ischemic bladders than in control bladders at an increase in length between 6 and 12 mm. In both control and ischemic bladders, active tension was highest at a 16-mm increase in length. Bladders subjected to unilateral or bilateral ischemia for 1 or 2 weeks demonstrated impaired contractile responses to field stimulation, bethanechol, ATP and KCl. There were no differences in contractile strength between muscle specimens obtained from the ipsilateral or contralateral sides of unilateral ischemic bladders. Our findings showed that unilateral and bilateral ischemia inhibited the in vitro contractile strength of the detrusor muscle in response to intramural and pharmacologic stimulation. PMID- 8712085 TI - Effects of in vivo Ischemia on the infusion cystometry and in vitro whole bladder contractility of the rat. AB - Ischemia induced by atherosclerosis is a common cause of organ failure in the elderly. We investigated the effects of in vivo ischemia created by ligation of the internal iliac arteries on the parameters of in vivo infusion cystometry under urethane anesthesia and on in vitro whole bladder contractility of the rat. Bladder weight significantly increased after ischemia for 14 days. Infusion cystometry demonstrated that in the ischemic bladders the capacity increased, the voiding pressure decreased, and the volume of residual urine increased, which resulted in deteriorated voiding efficacy. The in vitro whole bladder contractility to field stimulation, bethanechol, ATP, and KCl was reduced by ischemia. The passive pressure increased as the bladder volume enlarged and the bladder compliance once decreased by ischemia on the 7th day, but increased on the 14th day. In an active volume-pressure relationship study the peak response was decreased by ischemia. The volume at which response reached a peak value shifted to a larger volume 14 days after surgery. In conclusion, ischemia impaired in vivo rat detrusor power to empty. Since detrusor contractility in vitro decreased in response to various kinds of stimulation, this deteriorated bladder function was supposed to be caused by muscle degeneration. PMID- 8712086 TI - [A case of solitary metastasis from renal cell carcinoma to the thyroid gland]. AB - A case of solitary metastasis with renal cell carcinoma to the thyroid gland is presented. The patient was a 54-year-old man found to have an abnormal mass in the neck. He had a past history of radical nephrectomy orignating from the right renal cell carcinoma 5 years earlier (pT2N0M0, G2 > 3, alveolar type, clear cell subtype). Ultrasonography revealed a tumor mass in the right hemithyroid gland. Right hemithyroidectomy was performed on April 19, 1995. Histopathologically, the removed thyroid tumor showed clear cell carcinoma. The possibility of a primary thyroid tumor was ruled out by immunohistochemical thyroglobulin staining, and the present case was thus diagnosed as of metastatic thyroid tumor of renal cell carcinoma. The present case is the 12th case of thyroid solitary metastasis of renal cell carcinoma reported in Japan to date. PMID- 8712087 TI - [Renal cell carcinoma accompanied with ossification: a case report]. AB - Renal cell carcinoma accompanied with ossification reported in Japan is very rare. We report a case of renal cell carcinoma with ossification pathologically in a 61-year-old woman. She suddenly presented with hypercalcemia that caused acute renal failure, 6 months after the operation and died shortly thereafter. We discuss the relationship between the tumor and the hypercalcemia. PMID- 8712088 TI - [Ureteral malacoplakia: a case report]. AB - Malacoplakia is a condition which occurs most often in the urogenital system, but ureteral malacoplakia is rare. A case of ureteral malacoplakia is presented. The patients was a 54-year-old woman admitted to our hospital with high fever as the chief complaint. Laboratory data suggested urinary tract infection. Escherichia coli were isolated from the urine. Ultrasonography indicated left hydronephrosis and excretory urography showed reduced function of the left kidney. From retrograde pyelography, obstruction of the left distal ureter was indicated. A ureteral carcinoma was suspected and nephroureterectomy was thus carried out. The histopathological diagnosis was malacoplakia. The patient is in good health without any indication of the disease at 1 year 9 months following surgery. PMID- 8712089 TI - [A case of localized amyloidosis of the ureter]. AB - A case of localized amyloidosis of the ureter is reported. The patient was a 49 year-old female whose chief complaint was macrohematuria. Roentogenographic examination showed left hydronephrosis due to stenosis of left middle ureter. Left nephrouretectomy with cuff was performed with a diagnosis of the left ureteral tumor, and pathological examination revealed localized amyloidosis of the left ureter. Localized amyloidosis of the ureter is a rare lesion, and this is the twenty-first case reported in the Japanese literature. Review of the literature revealed that it is difficult to differentiate this lesion from other ureteral tumors by roentgenographic examination, and it is important to perform preoperative or intraoperative biopsy of ureteral tumors if benign diseases cannot be ruled out. PMID- 8712090 TI - [Diagnostic magnetic resonance imaging (MRI) in fracture of the penis: a case report]. AB - A 23-year-old man was admitted to our hospital complaining of swelling and pain in the penis. Examination revealed fracture of the penis. The tear in the tunica albuginea was not palpable on physical examination of the penis, but magnetic resonance imaging (MRI) revealed the exact site of the tear in the tunica albuginea. MRI was very useful in localizing the site of fracture. PMID- 8712091 TI - [Phase I study of bicalutamide (Casodex), a nonsteroidal antiandrogen in patients with prostatic cancer]. AB - A phase I study (open trial) of bicalutamide (Casodex), a non-steroidal antiandrogen, was conducted on 16 patients with prostatic cancer (stage C to D). The patients were given 10, 30, 50, 80 or 100 mg of bicalutamide orally daily for 12 weeks. Adverse reactions were observed in 8 out of 16 patients, but almost all were mild. Breast pain, gynecomastia and hot flushes were observed in 6 patients. Adverse reactions regarding liver function tests were observed in 3 patients. These were increased glutamic-oxalacetic transaminase (GOT), glutamic-pyruvic transaminase (GPT), alkaliphosphatase (AL-P) or gamma guanosine 5'-triphosphate (gamma-GTP). However, during or after the treatment period the elevated values were reversed to the pretreatment level. In terms of efficacy, anti-tumor effect was observed in 1 or 2 patients at each dose. Serum concentrations of luteinizing hormone (LH), follicle stimulating hormone (FSH), testosterone and estradiol increased during treatment. Plasma concentrations of the R (-) enantiomer, which has antiandrogenic activity, reached the steady state 6-8 weeks after the initiation of treatment; its apparent plasma elimination half-life observed following repeated administration was 8.4 +/- 1.1 days. In conclusion, bicalutamide (10-100 mg od) is considered to be tolerated well enough to be administered to patients with prostatic cancer and has shown evidence of anti tumor effect. PMID- 8712093 TI - [Prognostic factors of renal cell carcinoma: a multivariate analysis]. AB - In a series of 104 patients with renal cell carcinoma, a variety of clinical and histological factors were analyzed to establish their value as prognostic factors. In a univariate analysis (log-rank test), the presence or absence of symptoms, tumor size, pT, N, M, pV categories, TNM stage, Robson stage, nuclear grade and cell type were significantly associated with cause-specific survival, while age, sex and tumor side were not. In a multivariate analysis using Cox's proportional hazard model of 73 informative cases, only N (0 vs. 1-3) and M (0 vs. 1) categories of stage and nuclear grade (1-2 vs. 3) proved to be independent prognostic markers. PMID- 8712092 TI - [Clinical early phase II study of bicalutamide (Casodex) in patients with prostatic cancer]. AB - To investigate the efficacy and safety of bicalutamide (Casodex) with its clinically recommended dose, the randomized early phase II study was performed in 124 patients with prostatic cancer (stage C, D). The patients were given 50, 80 or 100 mg of bicalutamide orally once a day in fixed doses for 12 weeks; 122 patients were eligible for evaluation. The overall response rate was 50.0% (20/40), 61.0% (25/41) and 53.7% (22/41) in the 50 mg, 80 mg and 100 mg groups, respectively. The response rate in prostate lesion, bone and lymph node metastases was slightly higher in the 80 mg group than in the 50 mg and 100 mg groups. The proportion of patients showing a response with regard to serum PSA (CR and PR) was 84.2, 92.7 and 97.6% in the 50, 80 and 100 mg groups, respectively. The incidence of adverse reactions was 65.0, 61.0 and 61.0% in the 50, 80 and 100 mg groups, respectively, and there was no significant difference in overall safety rating in the three groups. Frequent adverse reactions were gynecomastia and breast pain. Only one patient in the 80 mg group was withdrawn due to shortness of breath. Serum concentrations of LH, testosterone and estradiol increased significantly after treatment. Bicalutamide was concluded to be effective and well tolerated in patients with prostatic cancer, and its recommended dose was 80 mg once daily. PMID- 8712094 TI - [A clinical study of renal pelvic and ureteral cancer associated with bladder cancer]. AB - We reviewed 45 cases of transitional cell carcinoma of the renal pelvis and ureter with reference to the coexistence or subsequent development of bladder cancer. Bladder cancer was associated with an upper urinary tract neoplasm in 26 of the 45 cases. The 5-year survival rate for coexistence of bladder cancer was 56% and that for subsequent bladder cancer was 65.6%. The 5-year survival rate for 19 cases unrelated with bladder cancer was 46.7%. Therefore, there was no significant difference among the three groups. As to degree of the malignancy of the renal pelvis and ureter, the 5-year survival rate was 73.7% for G1 and G2 and 26.2% for G3. As to the depth of invasion, of the renal pelvis and ureter the 5 year survival rate was 71.8% and 31.1% in the patients with stage of T1, T2 and T3, T4. The prognosis of cancer of the upper urinary tract depended on the degree of the malignancy, and the depth of invasion. Ninety two percent of subsequent bladder cancer was detected within 2 years after resection of the primary cancer. PMID- 8712095 TI - [HLA antigens in patients with testicular germ cell tumors]. AB - The role of HLA antigens in the etiology of testicular germ cell tumor has been suggested previously. Several attempts have been made to establish associations of HLA antigens in patients with testicular cancers, but have yielded inconsistent results. We studied the frequencies of HLA antigens by serological typing and DNA typing and examined the association between testicular germ cell tumors and HLA antigens. The serological expression of HLA-A, -B, -C, -DR and -DQ antigens was analyzed in 23 patients with germ cell cancers of the testis. The findings indicated a trend towards an increase in the antigen DQ4 (p < 0.05). The 13 patients were typed for HLA DRB1 and DQB1 allele by PCR-RFLP analysis. The findings revealed an increased frequency of DRB1 0405 (p < 0.01) and DQB1 0401 (p < 0.01). PMID- 8712097 TI - A negative transthoracic echocardiogram obviates the need for transesophageal echocardiography in patients with suspected native valve active infective endocarditis. AB - We studied 134 patients with suspected native valve infective endocarditis who underwent transthoracic and transesophageal echocardiography. Our data suggest that in patients without prosthetic valves who have a technically adequate negative transthoracic echocardiogram, transesophageal echocardiography is unlikely to be of incremental benefit in diagnosing endocarditis. PMID- 8712096 TI - Metaanalysis of five reported studies on the relation of early coronary patency grades with mortality and outcomes after acute myocardial infarction. AB - Coronary patency has been used as a measure of thrombolysis success after acute myocardial infarction (AMI). The Thrombolysis in Myocardial Infarction (TIMI) study grading scale for coronary perfusion has gained wide acceptance, but the significance of individual grades on clinical outcome has not been adequately tested. We hypothesized that optimal outcomes would be achieved only with early (and maintained) TIMI grade 3 (complete) perfusion compared with TIMI grade 2 (partial perfusion, previously classified as a reperfusion success) or grades 0 or 1 (occluded arteries). Five recent, angiographically controlled, prospectively performed studies of thrombolysis in AMI were identified, representing 3,969 patients. Odds ratios for mortality by early perfusion grades were calculated using the Mantel-Haenszel test and combined in a weighted fashion. Results for selected clinical and laboratory outcomes by patency grade were also assessed. Overall, mortality averaged 8.8% for TIMI grade 0/1, 7.0% for grade 2, and 3.7% for grade 3 perfusion. The odds ratio (OR) for early mortality was substantially reduced for grade 3 versus <3 perfusion (OR = 0.45, confidence interval [CI] 0.34 to 0.61, p <0.0001). In pairwise comparisons, grade 3 was clearly superior to grade 2 (OR = 0.54, CI) 0.37 to 0.78, p = 0.001) as well as grades 0/1 (OR = 0.41, CI 0.30 to 0.56, p <0.0001). Acute and convalescent ejection fraction, regional wall motion, time to enzyme peaks (creatine kinase [CK], creatine kinase myocardial bond [CK-MB]), peak enzyme levels [CK, lactate dehydrogenase [LDH], LDH-1), and risk of heart failure were each significantly less in patients achieving grade 3 than grade 2 (or lower grades) perfusion. Results were observed despite the frequent use of interventions after angiography. This meta-analysis demonstrates that early and complete (grade 3) flow is associated with superior survival and clinical outcome; grade 2 perfusion results in an inferior outcome, closer to that of an occluded than an open artery. The goal of reperfusion strategies should be early and maintained TIMI grade 3 perfusion. PMID- 8712098 TI - Use of biplane transesophageal echocardiography as the only imaging technique for percutaneous balloon mitral commissurotomy. AB - Percutaneous balloon mitral commissurotomy with the Inoue technique was attempted in 9 patients with rheumatic mitral stenosis under the sole guidance of biplane transesophageal echocardiography and was completed in 8 patients. This preliminary experience points to the feasibility and relative safety of performing percutaneous balloon mitral commissurotomy under the sole guidance of biplane transesophageal echocardiography. PMID- 8712099 TI - Assessment of mean left atrial pressure from the left ventricular pressure tracing in patients with cardiomyopathies. AB - The direct measurement of mean left atrial pressure is difficult and the ability to assess mean left atrial pressure from the left ventricular (LV) pressure tracing obtained at cardiac catheterization is clinically useful. The current study describes the limitations of using LV pre-a-wave pressure or LV end diastolic pressure to estimate mean left atrial pressure and describe another index, mean LV diastolic pressure, which provides a better assessment of mean left atrial pressure. PMID- 8712100 TI - Use of interlocking detachable coils in embolization of coronary arteriovenous fistulas. AB - Thirteen patients underwent transcatheter closure of coronary artery fistulas using interlocking detachable coils alone or combined with fibered coils. Complete occlusion was achieved in 92% of patients; although inadvertent embolization of coils occurred in 23% of patients, all these coils were easily retrieved using snares. PMID- 8712101 TI - Radiation-induced cardiovascular dysfunction. AB - Coronary artery disease and valvular dysfunction are long-term complications of mediastinal irradiation. We describe 3 patients who underwent successful combined coronary artery bypass grafting and valve replacement for symptoms related to radiation-induced coronary artery and valvular disease. PMID- 8712102 TI - Immunofluorescence analysis of trihexosylceramide accumulated in the hearts of variant hemizygotes and heterozygotes with Fabry disease. AB - An immunofluorescence method was applied to detect trihexosylceramide accumulated in the cardiac tissues from a variant hemizygote and a heterozygous female with Fabry disease, the incidence of which had been suspected to be high. PMID- 8712103 TI - Assessment of heart rate variability by using different commercially available systems. AB - The results of heart rate variability analysis of the same Holter tape by using 4 different commercially available systems are statistically incomparable. This might have important implications when projecting and evaluating clinical trials. PMID- 8712104 TI - Standardization of coronary vasomotor tone with intracoronary nitroglycerin. AB - Maximal, reproducible, and thus "standardized" dilation of epicardial coronary arteries can be easily achieved with intracoronary bolus administration of 0.1 mg nitroglycerin without considerable decrease in blood pressure. The addition of other nitrocompounds or calcium antagonists cannot increase coronary dilation after nitroglycerin, but may be hampered by adverse effects. PMID- 8712105 TI - Transaortic balloon valvoplasty of the pulmonary valve. AB - An unconventional transaortic to transductal approach was performed to perforate and dilate the pulmonary valve in pulmonary atresia. Ductal arteriosus patency was maintained by prostaglandin. PMID- 8712106 TI - Fluvastatin versus pravastatin. PMID- 8712107 TI - Uncomplicated successful percutaneous transluminal coronary angioplasty does not affect cardiac troponin T plasma concentrations. PMID- 8712108 TI - Congestive heart failure and atrial natriuretic factor: a room for endothelins? PMID- 8712109 TI - Effects of habitual caffeine consumption on ambulatory blood pressure. PMID- 8712110 TI - Editorial: combination therapy for systemic hypertension (15 September 1995). PMID- 8712111 TI - Fibrinogen levels in women having coronary angiography. AB - Fibrinogen has emerged as a risk factor for coronary artery disease in men that equals cholesterol in importance. It is known to play an important role in reparative processes, and evidence is accumulating that fibrinogen/fibrin accumulates at the site of minimal vascular injury. Fibrinogen contributes significantly to blood viscosity and its adherence to endothelium may mediate progression of atheromatous lesions. This study was designed to examine a number of markers of risk in a consecutive series of cardiology patients undergoing coronary catheterizations over a 15-month period. This article examines the level of fibrinogen in relation to the number of reported coronary stenoses and disease severity in a series of Caucasian female patients (n = 101). Women were classified as diseased if they had at least 1 lesion > or = 25% in the coronary anatomy and nondiseased if they had no lesions > or = 25%. The number of reported lesions correlates significantly with fibrinogen levels (r = 0.36, p = 0.0002). Women with fibrinogen levels > or = 283 mg/dl had a 3.2-fold increased risk (95% confidence interval 1.2 to 9.1) of having at least 1 stenosis > or = 25% after adjusting for age and diabetic status. Smoking and body mass index did not differ by disease status and thus did not confound the finding. Mean fibrinogen levels showed a progressive positive association with increasing clinically defined vessel involvement (stenosis > or = 50%). PMID- 8712112 TI - Influence of acute myocardial infarction location on in-hospital and late outcome after primary percutaneous transluminal coronary angioplasty versus tissue plasminogen activator therapy. AB - In the Primary Angioplasty in Myocardial Infarction trial, 395 patients with acute myocardial infarction (AMI) were prospectively randomized to tissue plasminogen activator (tPA) or primary percutaneous transluminal coronary angioplasty (PTCA). In 138 patients with anterior wall AMI, in-hospital mortality was significantly reduced by treatment with PTCA compared with tPA (1.4% vs 11.9%, p = 0.01). PTCA also resulted in lower rates of death or reinfarction (1.4% vs 18.0%, p = 0.0009), recurrent myocardial ischemia (11.3% vs 28.4%, p = 0.01), and stroke (0.0% vs 6.0%, p = 0.037) in anterior wall AMI. The independent beneficial effect of treatment with primary PTCA rather than tPA in anterior wall AMI was confirmed by multivariate analysis and interaction testing. The in hospital mortality of 257 patients with nonanterior wall AMI was similar after PTCA and tPA (3.2% vs 3.8%, p = 0.82). Compared with tPA, however, primary PTCA resulted in a markedly lower rate of recurrent myocardial ischemia (9.7% vs 27.8%, p = 0.0002), fewer unscheduled catheterization and revascularization procedures, and a shorter hospital stay (7.0 vs 8.6 days, p = 0.01) in nonanterior wall AMI. Thus, compared with tPA, primary PTCA in patients with anterior wall AMI results in significantly improved survival, with lower rates of stroke, reinfarction, and recurrent myocardial ischemia. In nonanterior wall AMI, treatment with PTCA and tPA results in similar early mortality, although PTCA treated patients have a more stable hospital course characterized by reduced recurrent ischemia, fewer subsequent invasive procedures, and earlier discharge. PMID- 8712113 TI - Relation between perfusion defects on stress technetium-99m sestamibi SPECT scintigraphy and the location of a subsequent acute myocardial infarction. AB - Although the presence of perfusion defects on stress myocardial perfusion imaging has been shown to correlate with future cardiac events, including acute myocardial infarction (AMI), it is unknown whether the location of the AMI can be predicted. Therefore, for 25 patients who had an AMI following a stress technetium-99m sestamibi single-photon emission computed tomographic (SPECT) imaging study and whose infarct location could be determined, the territory of infarction was correlated with the location of previous myocardial perfusion defects. A SPECT perfusion defect had been present in 24 patients (96%). The AMI occurred in territories that showed a reversible defect in 14 patients (56%), whereas 3 infarctions (12%) were in territories that revealed a fixed defect, and 8 infarctions (32%) were in territories that had not shown a defect on prior SPECT imaging. Whereas the incidence of infarction in territories with a reversible defect was highest at 14 of 26 (54%), the incidence of infarction in territories with a fixed defect was 3 of 7 (43%), and in territories with no defect was 8 of 42 (19%) (p = 0.011). Neither the time interval between SPECT imaging and infarction, nor the perfusion defect severity, was related to the correlation between perfusion defect and infarct location. Thus, although AMI occurs most often at the site of previous perfusion defects, reversible or fixed, a substantial percentage occur in territories without a perfusion defect. These findings suggest that abnormalities on SPECT perfusion imaging, although they serve as markers of significant coronary disease and increase the likelihood of infarction, do not always predict the exact location of infarction. PMID- 8712114 TI - Transcatheter therapy of thrombotic-occlusive lesions in saphenous vein grafts. AB - Saphenous vein graft (SVG) disease, a form of accelerated atherosclerosis, remains a therapeutic conundrum. The use of stents after excluding the presence of thrombus has proved highly successful at short- and long-term follow-up. We report on 60 severely symptomatic patients with multiple subtotal and total thrombotic SVG occlusions who were treated with a combination of intragraft urokinase-verapamil infusion and insertion of multiple biliary stents. Stent deployment had a 100% success rate. No case of clinical subacute thrombosis was registered, and major in-hospital complications were uncommon (<1%). The clinical outcome was encouraging, with a 12-month event-free survival rate of 87% in the 57 evaluable patients. This method of therapy appears to be highly successful in the treatment of thrombus-containing occlusive SVG disease, in preventing the "no reflow" phenomenon, and in lessening the incidence of periprocedural non-Q-wave myocardial infarction. PMID- 8712115 TI - Myocardial beta-adrenoceptor density and plasma catecholamines in syndrome X. AB - Recent research has cast doubt on the ischemic hypothesis of etiology of syndrome X (anginal pain, ischemic-like changes in the stress electrocardiogram, but normal coronary arteriogram). Abnormalities of pain perception have been shown and abnormal sympathetic nervous system activation has also been implicated. The aim of this study was to test the hypothesis that downregulation of myocardial beta adrenoceptors is demonstrable in patients with syndrome X. Such downregulation would be consistent with raised myocardial catecholamine concentrations. We performed positron emission tomography with (11)C-CGP-12177 to measure beta-adrenoceptor density. Plasma catecholamines were sampled simultaneously and assayed using high-performance liquid chromatography. Twenty syndrome X patients (11 female, age 57 +/- 9 SD years, range 33 to 69) and 18 matched controls (9 women, age 50 +/- 13 years, range 25 to 65; p = NS vs patients) were studied. Myocardial beta-adrenoceptor density did not differ between syndrome X patients and controls: 8.0 (1.9) pmol/g for patients versus 8.3 (2.1) pmol/g for controls; p = 0.62. No differences were found between patients and controls for plasma norepinephrine (2.82 [1.07] and 2.76 [1.18] nM, respectively; p = 0.89) or for epinephrine (0.29 [0.14] and 0.30 [0.20] nM, respectively; p = 0.84). In patients with syndrome X, beta-adrenoceptor density is normal and, by inference, myocardial catecholamines would also be normal. This weakens the case for a generalized enhancement of sympathetic activation in this disorder, although increased sympathetic reactivity during actual episodes of chest pain remains a possibility. PMID- 8712116 TI - Comparison of procainamide and lidocaine in terminating sustained monomorphic ventricular tachycardia. AB - Efficacy of procainamide and lidocaine in terminating spontaneous monomorphic ventricular tachycardia (VT) was assessed in a randomized parallel study. Patients with acute myocardial infarction and those with poor hemodynamic tolerance of VT were excluded. Procainamide 10 mg/kg was given intravenously with an injection speed of 100 mg/min, and lidocaine was administered at an intravenous dose of 1.5 mg/kg in 2 minutes. Fourteen patients were randomized to lidocaine and 15 to procainamide. Termination occurred in 3 of 14 patients after lidocaine and in 12 of 15 patients after procainamide (p <0.01). Procainamide stopped 8 of 11 VTs not responding to lidocaine, and lidocaine stopped 1 of 1 not responding to procainamde. Of a total of 41 VT episodes, 4 of 15 responded to lidocaine and 20 of 26 to procainamide (p <0.01). Because of VT recurrences, 16 patients could be studied repeatedly with drugs given in the reversed order. This resulted in a total of 55 trials of 79 drug injections. Lidocaine terminated 6 of 31 VTs and procainamide 38 of 48 (p <0.001). The protocol was stopped in 4 cases because of adverse effects. A comparison of the QRS width and QT interval before and at the end of the injection revealed significant lengthening of these values after procainamide but no change after lidocaine. In conclusion, procainamide is superior to lidocaine in terminating spontaneously occurring monomorphic VT. PMID- 8712117 TI - Electrophysiologic effects and predictors of success of combination therapy with class Ia and Ib antiarrhythmic drugs for sustained ventricular arrhythmias. AB - Antiarrhythmic drugs remain the first line of therapy in patients with sustained ventricular arrhythmias. Although success with class Ia antiarrhythmic medications has been limited, there is evidence that the addition of a class Ib agent may improve results. A total of 110 consecutive patients referred for electrophysiologic evaluation who had inducible sustained ventricular arrhythmias resistant to a class Ia agent underwent repeat electrophysiologic study after the addition of a class Ib drug. Patients with ejection fraction >40% and ventricular fibrillation inducible in the baseline study had an 80% response rate, whereas those with inducible ventricular tachycardia and ejection fraction < or = 40% responded 11% of the time. Responders demonstrated marked prolongation of ventricular refractoriness and slight shortening of the QRS, whereas nonresponders had QRS prolongation and a more modest increase in ventricular refractoriness. Thus, the efficacy of class Ia/Ib combination therapy in patients with inducible sustained ventricular arrhythmias refractory to a class Ia drugs alone can be predicted by baseline variables. Marked prolongation of ventricular refractoriness in the absence of QRS prolongation appears to be a key factor in the success of this combination. PMID- 8712118 TI - Evaluation of blood pressure response to the combination of enalapril (single dose) and diltiazem ER (four different doses) in systemic hypertension. AB - Angiotensin-converting enzyme inhibitors and calcium antagonists are 2 classes of antihypertensive agents frequently used either as monotherapy or in combination. A 6-week, multicenter, randomized, double-blind, placebo-controlled trial was conducted in essential hypertensive patients (diastolic blood pressures [BP] when seated, 95-115 mm Hg) to evaluate the efficacy, dose response, and safety profiles of enalapril combined with a new once-daily formulation of diltiazem. BP and heart rate were measured at 5 (peak) and 24 (trough) hours after dose to assess the efficacy and pharmacodynamic profile of the combination given once daily. There were 336 patients randomly assigned to either enalapril 5 mg plus 1 of 4 dose levels of diltiazem ER (60, 120, 180, or 240 mg), enalapril 5 mg alone, or placebo. Each combination dose level produced statistically significant reductions (p <0.05) in trough diastolic BP when seated, compared with placebo ( 6.8, -8.3, -10.1, and -10.3 mm Hg for the diltiazem ER doses of 60, 120, 180, and 240 mg, respectively). The 3 highest combination dose levels resulted in statistically significant (p <0.05) decreases in trough diastolic BP when seated, compared with placebo. There was a significant (p <0.001) linear dose-response relation. A trough-to-peak ratio > or = 0.5 was shown for the 3 highest combination doses. Drug-related adverse events were seen in 8.9% to 19% of the combination patients, 14.3% of the enalapril patients, and 8.6% of the placebo patients. The frequency and type of adverse events were those currently noted with each drug studied when used as monotherapy. PMID- 8712119 TI - Antihypertensive treatment in hypertensive patients with normal left ventricular mass is associated with left ventricular remodeling and improved diastolic function. AB - Antihypertensive therapy in hypertensive patients with left ventricular (LV) hypertrophy causes hypertrophy regression and improved diastolic filling. Whether similar changes occur in hypertensive patients with diastolic dysfunction and no hypertrophy is unknown. We determined the effect of antihypertensive therapy of LV geometry and function in hypertensive patients without hypertrophy. In 18 mild to moderate hypertensive patients without significant hypertrophy, baseline echocardiograms and rest and exercise and radionuclide angiograms were performed. Subjects were treated for 8 to 12 months with the calcium channel blocker felodipine and then restudied 2 weeks after treatment withdrawal. Blood pressure normalized with treatment (165 +/- 22/98 +/- 9 to 128 +/- 12/80 +/- 5 mm Hg, p <0.001) and returned to pretreatment levels after therapy withdrawal. Rest ejection fraction and peak oxygen consumption and cardiac outputs were unchanged after treatment, but rest peak filling rate increased (2.63 +/- 0.57 to 3.11 +/- 0.95 end-diastolic volume/second, p <0.05). Ejection fraction increased with exercise only after treatment (64 +/- 5% at rest to 71 +/- 8% at peak exercise, p <0.05). LV mass index was unchanged (97 +/- 18 to 101 +/- 23 g/m2), but relative wall thickness declined (0.41 +/- 0.05 to 0.37 +/- 0.05) and LV end-diastolic dimension increased (4.9 +/- 0.4 to 5.2 +/- 0.4, p = 0.01). Blood pressures control in hypertensive patients without hypertrophy leads to improved peak filling rates and remodeling with decreased relative wall thickness. Improved diastolic function can occur without alterations in LV mass. PMID- 8712120 TI - Comparison of enalapril versus nifedipine to decrease left ventricular hypertrophy in systemic hypertension (the PRESERVE trial). AB - The PRESERVE (Prospective Randomized Enalapril Study Evaluating Regression of Ventricular Enlargement) study is designed to provide a definitive test of the ability of enalapril to achieve greater left ventricular (LV) mass reduction than nifedipine GITs (gastrointestinal treatment system) by a degree that would be prognostically meaningful on a population basis (10 g/m2). To achieve this goal, an ethnically diverse population of 480 men and women with essential hypertension and increased LV mass of screening echocardiography will be enrolled at clinical centers on 4 continents and studied by echocardiography at baseline and after 6 and 12 months' randomized therapy. Blinded readings of echocardiograms at a central laboratory will provide systematic information about treatment effects on LV structure, wall motion, and Doppler blood flow. The study power is at least 90% to test the primary hypotheses that enalapril will induce greater normalization of LV mass and diastolic filling than nifedipine. After the 1-year echocardiographic trial, the study population will be followed 3 more years to test the hypothesis that a reduction in LV mass, independent of blood pressure lowering, is associated with a reduction in the risk of morbid and fatal cardiovascular events. PMID- 8712121 TI - Normalization of left ventricular nonuniformity late after valve replacement for aortic stenosis. AB - The aim of the present study was to evaluate nonuniformity in pressure overload hypertrophy due to aortic stenosis. Twenty patients were included in the present analysis. Ten patients with severe aortic stenosis were studied preoperatively as well as early (21 +/- 8 months) and (89 +/- 21 months) after aortic valve replacement (AVR) using left ventricular biplane angiograms, high-fidelity pressure measurements and endomyocardial biopsies. Ten normal subjects served as controls. LV systolic function was assessed from biplane ejection fraction, and diastolic function from the time constant of relaxation, the peak filling rate and the constant of myocardial stiffness. Nonuniformity was evaluated from the coefficient of variation of the time to end-systole (systolic asynchrony) and peak filling rate (diastolic asynchrony) of 12 regions in right anterior oblique and left anterior oblique projection. Ejection fraction was comparable in patients with aortic stenosis and in controls, whereas preoperatively diastolic dysfunction with prolonged relaxation and increased stiffness was present in patients with aortic stenosis and was normalized late after AVR. LV systolic asynchrony was present (>25D of controls) in 7 and diastolic asynchrony in 10 of 10 patients with aortic stenosis. Early as well as late after AVR systolic asynchrony was normalized in 9 of 10 patients. Diastolic asynchrony was present early AVR in all but one patient, although there was a significant improvement with respect to the preoperative evaluation. Late after AVR there was a normalization of diastolic asynchrony in 9 of 10 patients with aortic stenosis. Thus, it is concluded that systolic asynchrony is normalized early after AVR probably due to its load-sensitivity, whereas diastolic asynchrony persists probably due to residual LV hypertrophy with increased interstitial fibrosis and myocardial stiffness. Late after AVR, diastolic asynchrony is normalized due to structural remodeling with regression of both myocardial hypertrophy and interstitial fibrosis. PMID- 8712122 TI - Eric Jeffrey Topol: a conversation with the editor. PMID- 8712123 TI - Lidocaine and sustained monomorphic ventricular tachycardia: fact or fiction. PMID- 8712124 TI - Low incidence of serotonin-induced occlusive coronary artery spasm in patients with recent myocardial infarction. AB - Intracoronary infusion of serotonin has been reported to induce varying degrees of coronary vasoconstriction in different coronary syndromes, but it has never been studied in patients after myocardial infarction. In patients with recent myocardial infarction, we found a low incidence (11%) of serotonin-induced occlusive spasm only in the infarct-related artery (IRA), and a significantly higher vasoconstriction in the distal segment of the IRA than in the same segment of the non-IRA. PMID- 8712125 TI - Different effects of acute administration of aminophylline and nitroglycerin on exercise capacity in patients with syndrome X. AB - To investigate the effects of the acute administration of aminophylline and nitroglycerin on effort ischemia, 20 patients with syndrome X underwent 3 bicycle exercise tests after sublingual nitroglycerin (0.3 mg) and after 90 minutes of oral administration of aminophylline (400 mg). Compared with the basal test, only aminophylline induced a significant increase in the time to ischemic threshold and to angina; these findings support the potential therapeutic role of this adenosine receptor blocking agents and suggest a possible role of "steal phenomenon" in the pathogenesis of effort angina in patients with syndrome X. PMID- 8712126 TI - Comparison of presentation, treatment, and outcome of acute myocardial infarction in men versus women (the Myocardial Infarction Triage and Intervention Registry) AB - This study compared the presentation (symptoms and signs), treatment, and outcome of 1,097 consecutive patients (851 men and 246 women) from the Myocardial Infarction Triage and Intervention (MITI) Project Registry with confirmed acute myocardial infarction (AMI), all of whom were initially evaluated in the prehospital setting, met clinical criteria for possible thrombolysis, and were followed throughout their hospital course. Women were older than men and had a higher prevalence of known cardiovascular risk factors, including systemic hypertension and congestive heart failure. The presentation of AMI with respect to symptoms, delay, and hemodynamic and electrocardiographic findings was for the most part indistinguishable between mean and women. Women appeared "undertreated" early in the course of AMI and were half as likely as men to undergo acute catheterization, angioplasty, thrombolysis, or coronary bypass surgery (odds ratio 0.5 [0.3 to 0.7]). The risk for hospital mortality in women was almost twice that for men (odds ratio 1.95 [1.01 to 3.8]). Hospital mortality after AMI was also independently predicted by older age, early evidence of hemodynamic instability, and an intraventricular conduction abnormality on the initial electrocardiogram. Although similar in its presentation, AMI in women is not as aggressively treated, and results in a less favorable outcome than in men. Gender as well as nongender-specific risk factors are important in assessing risk and the likelihood of early intervention after AMI. PMID- 8712127 TI - Long-term influence of fibrinogen on initial and recurrent cardiovascular events in men and women. AB - Examination of the long-term relation of a single fibrinogen determination to initial and recurrent atherosclerotic cardiovascular events over 20 years of follow-up revealed a powerful and comparably independent impact on initial events in both sexes but an influence on recurrent events only in men. PMID- 8712128 TI - The results of atrial flutter ablation in patients with and without a history of atrial fibrillation. AB - To determine the impact of atrial flutter radiofrequency catheter ablation on recurrence of atrial flutter and atrial fibrillation, 32 patients with atrial flutter (18 with a history of atrial fibrillation) were followed for a mean of 8.6 months; atrial flutter has not recurred after 1 (26 patients) or 2 (5 patients) successful ablation procedures. Atrial flutter did not appear proarrhythmic for atrial fibrillation, with only 1 of 15 patients without a history of atrial fibrillation developing the arrythmia in the absence of an alcohol binge or cocaine use. PMID- 8712129 TI - Effects of electrode polarity on defibrillation thresholds in biphasic endocardial defibrillation. AB - The defibrillation thresholds of both first-phase polarities of a biphasic waveform were tested using an endocardial defibrillation electrode system. Despite differences in defibrillation thresholds in the individual patient, both tested electrode polarities lead to a comparable overall defibrillation efficacy. PMID- 8712130 TI - Natural history of aortic valve stenosis of varying severity in the elderly. AB - In a population sample of 501 persons aged 75 to 86 years, Doppler echocardiography uncovered moderate or severe aortic valve stenosis in 8.8% of women and 3.6% of men. Severe aortic valve stenosis predicted a four-fold-age- and sex-adjusted risk of death within 4 years of diagnosis, and mortality tended to be increased also with moderate lesions; mild aortic valve stenosis had a favorable outcome. PMID- 8712131 TI - Compression of digital coronary angiograms does not affect visual or quantitative assessment of coronary artery stenosis severity. AB - Digital coronary angiographic techniques are now widely used in many cardiac catheterization laboratories. However, the full potential of digital imaging technology remains to be achieved because of its enormous storage and exchange requirements. Compression of digital imaging data allows a reduction in the volume of data so that storage and transmission are more efficient and cost effective. Three angiographers reviewed the original and compressed formats of 96 coronary angiographic sequences in a blinded fashion to assess coronary lesion severity. Compression was achieved using the Joint Photographic Experts Group (JPEG) standard, which resulted in a compression ratio of approximately 15:1. The original format was reviewed in a blinded fashion a second time to assess for intraobserver variability of similar formats. Lesion severity was graded in quartiles. Coronary stenosis >50% was considered "significant." In parallel, the reproducibility of quantitative coronary angiographic (QCA) measurements of coronary artery dimensions was also evaluated. For the visual assessment of lesion severity in the compressed versus original formats, kappa=0.52, suggesting moderate agreement. When lesions were assessed as significant versus "insignificant," however, kappa=0.88, suggesting excellent agreement. In the 2 separate readings of the original data formats, kappa=0.44 for assessment of lesion severity by quartiles and kappa=0.72 for lesions assessed as significant versus insignificant. Analysis of the compressed versus original data sets using QCA resulted in an excellent correlation for the measurement of lesion severity (r=0.99). The correlation was equally strong when the original format was analyzed sequentially (r=0.98). Lossy JPEG (15:1) compression is a valid means for reducing storage and exchange requirements of coronary angiographic data. The variability in assessing lesion severity between the original and compressed formats is comparable to the reported variability in visual assessment of lesion severity in sequential analysis of cine film. PMID- 8712132 TI - Prognostic value of predischarge exercise testing, ejection fraction, and ventricular ectopic activity in acute myocardial infarction treated with streptokinase. AB - The relative importance of prognostic parameters that delineate left ventricular function, myocardial ischemia, and arrhythmogenic potential after thrombolytic therapy is not clear. This study investigated 112 patients with acute myocardial infarction who were treated with thrombolysis to determine the relative prognostic value of predischarge treadmill exercise testing, radionuclide ventriculography, and ambulatory electrocardiographic monitoring for ventricular ectopic activity. During a mean follow-up period of 18 months (range 6 to 30), 42 first cardiac events were recorded, consisting of 3 deaths, 6 reinfarctions, 16 bouts of unstable angina, 16 episodes of heart failure, and 1 arrhythmic event. Univariate analysis revealed ejection fraction, exercise time, and ventricular ectopic count of > or = 10/hour to be predictive of future cardiac events. Subsequent multivariate analysis showed ejection fraction (p <0.001) and exercise time (p=0.002 to have independent prognostic value, but ventricular ectopic activity did not provide additional information. Ventricular ectopic count > or = 10/hour was additionally predictive only when combined with either ejection fraction (R2=5.4%) or exercise time (R2=2.9%). Event-free survival analysis revealed hazard ratios for ejection fraction <40% and exercise time <7 minutes of 3.63 (p=0.001) and 2.16 (p=0.01), respectively. Although ejection fraction and exercise time were able to predict future episodes of heart failure, neither could adequately identify patients at risk of recurrent ischemic events. PMID- 8712133 TI - Prognostic value of plasma fibrinogen concentration in patients with unstable angina and non-Q-wave myocardial infarction (TIMI IIIB Trial) AB - Inflammation may play an important role in acute coronary syndromes. We studied the prognostic value of fibrinogen, an acute-phase protein directly involved in thrombotic process, measured serially in 1,473 patients with unstable angina and non-Q-wave myocardial infarction participating in the Thrombolysis in Myocardial Infarction IIIB trial. Overall, no association was found between baseline (pretreatment) fibrinogen and in-hospital (< or = 10 days) myocardial infarction (p=0.70) and death (p=0.64); however, patients with spontaneous ischemia (p=0.004) and the combined unsatisfactory outcome of death, myocardial infarction, and spontaneous ischemia (p=0.003) had higher fibrinogen concentrations than those without these events. This association was confined to patients with unstable angina. A baseline fibrinogen concentration > or = 300 mg/dl was associated with a modest trend toward an increased risk of death, myocardial infarction, or spontaneous ischemia (odds ratio 1.61, 95% confidence interval 1.02 to 2.52; p=0.04). Elevation of fibrinogen, a readily measurable acute-phase protein, at the time of hospital admission is associated with coronary ischemic events and a poor clinical outcome in patients with unstable angina. PMID- 8712134 TI - Usefulness of intracoronary stenting in acute myocardial infarction. AB - Data on the feasibility, safety, and clinical outcome of intracoronary stenting in acute myocardial infarction (AMI) are limited. This study examined the immediate angiographic results and the early and late outcomes in 32 patients who had stenting during AMI. Coronary angiograms recorded at the time of stenting were reviewed with quantitative measurements obtained on the "target" coronary lesion before and after stenting. Immediate angiographic success was achieved in 30 patients (94%). The minimal luminal diameter increased from 0.36 +/- 0.37 to 2.58 +/- 0.41 mm (p<0.0001). Two patients died in the hospital. Of the remainder, none had reinfarction or required bypass surgery, whereas 2 required repeat coronary angioplasty for recurrent ischemia. Although thrombus at the infarct related coronary lesion was initially detected in 41% of the patients, its presence was not associated with adverse procedural outcome. Only 1 patient had persistent thrombus after stenting, which resolved with intracoronary urokinase. At a mean follow-up of 6.1 +/- 4.1 months, there was 1 additional cardiac death, and no patient had AMI or required repeat coronary angioplasty or bypass; among the 29 survivors, 86% were free of angina. Thus, intracoronary stenting of the infarct-related artery in the setting of AMI is associated with excellent immediate angiographic success and a favorable clinical outcome, and remains an option even in the presence of thrombus. PMID- 8712135 TI - Trends and determinants of calcium antagonist usage after acute myocardial infarction (the GISSI experience). AB - In the last decade, several clinical trials in patients with, or recovering from, acute myocardial infarction (AMI) have evaluated the role of calcium antagonists in affecting patients' prognosis. Results have been disparate, with evidence of possible harm, no effect, or some benefit, depending on the agent used. We evaluated how the evidence from these trials has influenced the pattern of prescription of calcium antagonists and assessed the important determinants of use of these agents in patients after AMI. We analyzed retrospectively the prescription of calcium antagonists at discharge in all patients recovering from AMI enrolled in 3 large randomized clinical trials (Gruppo Italiano per lo Studio della Sopravvivenza nell' Infarto-1 [GISSI-1], GISSI-2, and GISSI-3) during the last 10 years. A progressive decrease in prescriptions for calcium antagonists was evident, from 47.2% in GISSI-1 to 35.1% in GISSI-2 to 19.0% in GISSI-3 (p<0.001). The presence of post AMI angina, history of hypertension, and occurrence of reinfarction were associated with a higher usage of calcium antagonists, whereas the use of beta blockers at discharge was a major independent negative determinant. Use of calcium antagonists for secondary prevention after AMI (i.e., without specific clinical indications for their use) decreased by approximately 60% (from 26.1% to 10.3%). The data indicate that the usage of calcium antagonists in GISSI studies has been strongly affected by the results of other large multicenter trials evaluating calcium antagonists. These agents are now prescribed in patients after AMI almost exclusively in the presence of specific indications such as systemic hypertension or angina. PMID- 8712136 TI - Influence of age on the prognostic importance of left ventricular dysfunction and congestive heart failure on long-term survival after acute myocardial infarction. TRACE Study Group. AB - The aim of this study was to assess the importance of congestive heart failure and left ventricular (LV) systolic dysfunction after an acute myocardial infarction (AIM) on long-term mortality in different age groups. A total of 7,001 consecutive enzyme-confirmed AMIs (6,676 patients) were screened for entry into the TRAndolapril Cardiac Evaluation (TRACE) study. Medical history, echocardiographic estimation of LV systolic function determined as wall motion index, infarct complications, and survival were documented for all patients. To study the importance of congestive heart failure and wall motion index independent of age, we performed Cox proportional-hazard models in 4 different age strata (< or = 55 years, 56 to 65 years, 66 to 75 years, and > 75 years). Patients in these strata had 1-year mortality rates of 5%, 11%, 21%, and 32%, respectively. Three-year mortality rates were 11%, 20%, 34%, and 55%, respectively. The risk ratios (and 95% confidence limits) associated with congestive heart failure in the same 4 age strata were 1.9 (1.3 to 2.9), 2.8 (2.1 to 3.7), 1.8 (1.5 to 2.2) and 1.8 (1.5 to 2.2), respectively. The risk ratios associated with decreasing wall motion index were 6.5 (3.6 to 11.4), 3.3 (2.3 to 4.6), 2.7 (2.2 to 3.4), and 2.1 (1.7 to 2.6), respectively. In absolute percentages, there was an excess 3-year mortality associated with congestive heart failure in the 4 age strata of 14%, 24%, 25%, and 28% respectively. The absolute excess in 3-year mortality associated with LV systolic dysfunction in the 4 age strata was 15%, 19%, 25%, and 21%, respectively. Thus, the relative importance of LV systolic dysfunction and congestive heart failure diminished with increasing age. However, the absolute excess mortality associated with congestive heart failure and LV systolic dysfunction was more pronounced in the elderly than in the young. PMID- 8712137 TI - Predictive value of lipid profile for salutary coronary angiographic changes in patients on a low-fat diet and physical exercise program. AB - In this study, 113 patients with modestly elevated levels of low-density lipoprotein cholesterol (<210 mg/dl) and coronary artery disease were randomized to an intervention group (n=56) or a control group (n=57). The intervention program consisted of daily exercise and a low-fat diet according to the American Heart Association's recommendation phase III; patients in the control group received "usual care" rendered by their private physician. After 1 year, complete data were available for all 92 patients (intervention: n=40; control: n=52) who underwent repeat coronary angiography. During the study course, patients in the intervention group showed an increase in apolipoprotein A-I(123 +/- 18 vs 129 +/- 20 mg/dl; p < 0.02) and apolipoprotein A-I/B (1.3 +/- 0.4 vs 1.5 +/- 0.4; p <0.01) and a decrease in apolipoprotein B (99 +/- 20 vs 89 +/- 18 mg/dl; p < 0.01), while apolipoprotein A-II remained unchanged (38 +/- 6 vs 38 +/- 6 mg/dl; p=NS). In the control group, there were no significant changes (apolipoprotein A I, 124 +/- 17 vs 128 +/- 13 mg/dl; apolipoprotein A-II, 38 +/- 6 vs 39 +/- 6 mg/dl; apolipoprotein B, 100 +/- 21 vs 99 +/- 16 mg/dl; apolipoprotein A-I/B, 1.3 +/- 0.3 vs 1.4 +/- 0.5; all p=NS). As previously reported, there was a significant retardation of progression in patients in the intervention group (progression 23%, no change 45%, regression 32%) compared with the control group (progression 48%, no change 35%, regression 17%) (p < 0.05). Although retardation of progression was significantly associated with an increase in apolipoprotein A I/B and a decrease in apolipoprotein B (p < 0.05), these gave way in multivariate analysis to changes in total cholesterol/high-density lipoprotein cholesterol, absolute levels of low-density lipoprotein cholesterol, and, in a subgroup of patients, to leisure-time physical activity (all p < 0.05). These data demonstrate that an intervention based on a low-fat diet and intensive physical exercise is capable of improving apolipoprotein levels, associated with retardation of progression of coronary artery disease. However, total cholesterol/high-density lipoprotein cholesterol and low-density lipoprotein cholesterol appear superior to apolipoproteins as metabolic markers for effective treatment in patients with coronary artery disease. PMID- 8712138 TI - Dobutamine stress echocardiography for noninvasive diagnosis of cardiac allograft vasculopathy: a comparison with angiography and intravascular ultrasound. AB - This study was performed to assess the value of dobutamine stress echocardiography for noninvasive diagnosis of cardiac allograft vasculopathy (CAV) compared with coronary angiography and intravascular ultrasound (IVUS) in 50 consecutive orthotopic heart transplant recipients. In 46 of 50 patients, a technically adequate echocardiogram could be obtained. Using a 16-segment model, a total of 675 segments were analyzed. At rest, wall motion abnormalities were found in 61 of 675 (9.0%) left ventricular segments in 15 of 46 patients. At maximal dobutamine stress, 103 of 675 segments (15.3%) had wall motion abnormalities (25 of 46 patients). Based on IVUS and angiographic findings, patients were allocated to 2 groups. Group I (n=18) had absent or only mild intimal hyperplasia (mean IVUS grade < or = 3.0 on a 6-grade scale). Group II (n=28) had moderate to severe intimal hyperplasia (mean grade > 3.0 with or without angiographic evidence of CAV. The prevalence of wall motion abnormalities was significantly higher in group II than in group I, both at rest (50 of 415 vs 11 of 270 coronary segments in 13 of 28 vs 2 of 18 patients) and during maximal stress (88 of 415 vs 15 of 270 coronary segments in 22 of 28 vs 3 of 18 patients). Quantitative M-mode echocardiography demonstrated decreased wall thickening in group II versus group I patients at maximal dobutamine dosage in the septum (48 +/- 18% vs 61 +/- 17%; p < 0.01) as well as in the left ventricular posterior wall (77 +/- 21% vs 96 +/- 21%; p <0.005). Regional myocardial dysfunction as assessed by dobutamine stress echocardiography was associated with IVUS evidence of moderate to severe intimal hyperplasia. Dobutamine stress echocardiography appears to be a feasible noninvasive method for detection of CAV in heart transplant recipients, which may reduce the need for routine coronary angiography. PMID- 8712139 TI - Evaluation of myocardial ischemia and infarction by signal-averaged electrocardiographic late potentials in children with Kawasaki disease. AB - We investigated myocardial ischemia and old myocardial infarction noninvasively using signal-averaged electrocardiographic late potentials (LPs) in patients with Kawasaki disease. Patients were divided into 4 groups: a noncoronary artery lesion group (n=136), a coronary artery lesion group (without myocardial ischemia and an old myocardial infarction; n=33), an ischemia group (n=16), and an old myocardial infarction group (n=13). Grouping was based on exercise thallium-201 myocardial scintigraphy, thallium-201 myocardial scintigraphy, exercise electrocardiography, coronary angiography, left ventriculography, and echocardiography. Signal-averaged electrocardiograms were recorded using a high resolution system. Values of filtered QRS duration (f-QRSd), root-mean-square voltage, and duration of low-amplitude signal were judged using our own body surface area-related criteria (n=205) to determine positive rates of LPs and sensitivities and specificities to ischemia and infarction. These data were also interpreted using published criteria for adults and compared with those interpreted by our criteria. Positive rates by our criteria were 0% in the noncoronary artery lesion group, 9.1% in the coronary lesion group, 56.3% in the ischemia group, and 69.2% in the old myocardial infarction group. However, using the criteria for adults, these values were 0%, 3.0%, 25%, and 46.2%, respectively. Sensitivities to ischemia and infarction using our criteria were significantly higher (56.3% and 69.2%) than those using the criteria for adults (p < 0.05). Moreover, specificities to ischemia and infarction were very high (93.4% and 93.5%, respectively) using our criteria, and there were no significant differences from specificities using the criteria for adults. Also, we examined the reproducibility of values of LPs and LP parameters. The values of filtered QRS duration showed a high reproducibility in both LP-positive and -negative groups, followed by low-amplitude signal and then root-mean-square voltage. The results of LP presence or absence showed 100% reproducibility for both the LP positive and -negative groups, supporting the utility of LPs for clinical applications. Thus, LPs provide useful information in a noninvasive manner for clarifying ischemia and infarction in patients with Kawasaki disease. PMID- 8712140 TI - Frequency of systemic microvascular dysfunction in syndrome X and in variant angina. AB - In addition to coronary vascular abnormalities, patients with syndrome X and variant angina often have systemic vascular symptoms. To determine whether these patients exhibit a generalized abnormality of vasoreactivity, we used high resolution ultrasound to compare flow responses and endothelial function in the brachial artery in 21 patients with syndrome X, 15 patients with variant angina, and 20 healthy controls. Arterial diameter was measured at rest, after reactive hyperemia (endothelium-dependent flow-mediated vasodilation), and after sublingual glyceryl trinitrate (endothelium-independent vasodilation). The magnitude of hyperemic flow response was measured after transient forearm occlusion. Flow-mediated dilation in the brachial artery did not differ among patients with syndrome X, variant angina, and controls (2.7 +/- 2.3%, 3.8 +/- 3.5%, and 4.2 +/- 3.0%). Endothelium-independent vasodilation in the brachial artery was similar in the 3 groups (16.0 +/- 7.2%, 12.7 +/- 4.6%, and 14.8 +/- 4.9%). Despite a considerable overlap, reactive hyperemia was lower in patients with syndrome X than in patients with variant angina and controls (342+/-86% vs 466+/-184% and 452+/-104%; p < 0.05). These findings indicate that a substantial proportion of patients with syndrome X have a systemic microvascular abnormality, whereas variant angina is predominantly a segmental disorder of conduit vessels. PMID- 8712141 TI - Survey of cardiac pacing and defibrillation in the United States in 1993. AB - A survey of implanters of permanent cardiac pacemakers and implanted cardioverter defibrillators (ICDs) in the United States during 1993 was conducted to identify present and changing patterns in indications for pacing implantation techniques, pacing-mode selection, follow-up, and opinions regarding pacing-and ICD-related issues. Five major pacemaker manufacturers also provided estimates of the numbers of pacemakers and ICDs implanted in the United States from 1990 through 1993. In 1993 approximately 133,000 new rhythm-management devices, including 112,000 primary pacing systems and 16,000 ICDs, were implanted, an increase of 18% for pacemakers and 113% for ICDs since 1990. In 1993, pacemaker implantations were performed by about 8,700 physicians working in 3,300 hospitals and 1,000 independent "surgi-centers." Since the last survey, which addressed pacing practices in 1989, respondents' use of dual-chamber pacemakers increased from 32% to 68% of the total, and adaptive-rate systems from 29% to 48%. Significant differences (p <0.05) were found among subcategories of implanters and among complications encountered in different circumstances. Surgeons tended be the older and more experienced implanters, but used a smaller proportion of active fixation leads, dual-chamber systems, and adaptive-rate pulse generators, and tended to rely more heavily on a pacemaker manufacturer's representative in operative and follow-up procedures. Complications were more common with bipolar leads, with leads implanted by means of an introducer, and with passive-fixation ventricular leads. The survey provided useful insights into trends and differences in pacemaker and defibrillator practice. Future surveys would be facilitated if a standardized implant registry such as that used in Europe were established in the United States. PMID- 8712142 TI - Prognostic value of left ventricular mass and geometry in systemic hypertension with left ventricular hypertrophy. AB - To determine the independent prognostic significance of left ventricular (LV) mass and geometry (concentric vs eccentric pattern) in hypertensive subjects with LV hypertrophy at echocardiography, 274 subjects were followed for up to 8.7 years (mean 3.2). All patients had systemic hypertension and LV mass > or = 125 g/body surface area (BSA) and underwent ambulatory blood pressure (BP) monitoring and echocardiography before treatment. Eccentric and concentric hypertrophy were defined by the ratio between LV posterior wall thickness and LV radius at telediastole <0.45 and > or = 0.45, respectively. Age, sex ratio, body mass index, office BP and serum glucose, cholesterol, and triglycerides did not differ between the groups with eccentric (n=145) and concentric (n=129) hypertrophy. Average 24-hour daytime, and nighttime systolic ambulatory BPs were higher in concentric than in eccentric hypertrophy (all p <0.01). LV mass was slightly greater in concentric than in eccentric hypertrophy (157 vs 149 g/BSA, p <0.05). Endocardial and midwall shortening fraction were lower in concentric than in eccentric hypertrophy (96.5% vs 106.0% of predicted and 71.4% vs 89.7% of predicted, respectively; both p <0.01). The rate of major cardiovascular morbid events was 2.20 and 3.34 per 100 patient-years in eccentric and concentric hypertrophy, respectively (log rank test, p=NS). Age >60 and LV mass above median (145 g/BSA) were significant adverse prognostic predictors, while LV geometry (eccentric vs concentric hypertrophy) and ambulatory BP were not. The event rates per 100 patient-years were 1.38 and 3.98, respectively, in the patients with LV mass below and above median (age-adjusted relative risk 2.70; 95% confidence interval [CI] 1.03 to 6.63; p=0.015). In hypertensive subjects with established LV hypertrophy, LV mass, but not its geometric pattern, provides important prognostic information independent of conventional risk markers including office and ambulatory BP. PMID- 8712144 TI - Ethnic differences in forearm vasodilator capacity. AB - Previous studies have demonstrated significant ethnic differences in left ventricular structure in both normotensive and hypertensive subjects. To determine if these differences in ventricular geometry are associated with differences in vascular structure, we measured the minimum forearm vascular resistance in 30 healthy young African-American adults and in 30 whites matched for age, gender, and blood pressure. Average daytime blood pressure was determined by ambulatory monitoring during a typical work day. Minimum forearm vascular resistance was measured by plethysmography after 10 minutes of forearm ischemia. Indexed left ventricular mass and relative wall thickness were measured by 2-dimensional-directed M-mode echocardiography. The mean (+/-SD) ambulatory pressure was 126 +/- 11/79 +/- 8 mm Hg in African-Americans and 126 +/- 11/79 +/- 7 mm Hg in whites. The 2 groups were similar in body mass index and in family history of hypertension. African-Americans had a higher minimum forearm vascular resistance than did whites (2.39 +/- 0.75 vs 2.03 +/- 0.55 mm Hg, p <0.05). There was a trend toward a greater left ventricular relative wall thickness in African Americans (0.38 +/- 0.07 vs 0.35 +/- 0.06, p=0.09). These results suggest that early vascular remodeling is present in African-Americans who do not have established hypertension, and that this ethnic difference in vascular structure is associated with a difference in ventricular geometry. PMID- 8712143 TI - Comparison of the effects of amlodipine and diltiazem on 24-hour blood pressure, plasma catecholamines, and left ventricular mass. AB - In 30 patients with mild to moderate essential hypertension and high-normal left ventricular (LV) mass, the effects of treatment for 6 months with amlodipine (5 to 10 mg every morning) versus diltiazem-sustained release (SR) (90 to 180 mg twice daily) on 24-hour blood pressure (BP), plasma catecholamines, and echocardiographic estimates of LV mass and function were evaluated. Both amlodipine and diltiazem caused stable, persistent BP reduction over 24 hours with no evidence for a "peak" effect. For a similar decrease in diastolic BP, amlodipine caused a significantly larger decrease in systolic BP. Amlodipine decreased BP by lowering total peripheral resistance, whereas diltiazem caused small decreases in both total peripheral resistance and cardiac index. Both calcium antagonists caused modest but significant decreases in supine and standing plasma catecholamines. LV wall thickness and LV mass decreased significantly over the 6 months of follow-up: -6 +/- 2 with diltiazem and -10 +/- 2 g/m2 with amlodipine. In patients taking amlodipine, the decrease in LV mass correlated significantly with the decrease in plasma norepinephrine. In contrast to rapid-acting calcium antagonists, both amlodipine and diltiazem-SR cause smooth BP control and an appropriate decrease in LV mass without activation of the sympathetic nervous system. PMID- 8712145 TI - A videodensitometric study of transmural heterogeneity of cyclic echo amplitude variation in human myocardium. AB - The aims of this study were: (1) to assess whether variations in cyclic echo amplitude might be detected across the human myocardium by videodensitometric analysis of images obtained with epicardial echocardiography; and (2) to explore the possible relation between cyclic gray level variation and left ventricular (LV) hypertrophy and function. Experimental studies show that transmural differences in contractile performance across the normal myocardium are paralleled by differences in the cyclic (diastolic-to-systolic) variation of myocardial echo amplitude. Thirty-three patients (aged 60 +/- 11 years) undergoing cardiac surgery were studied by intraoperative epicardial echocardiography. LV mass index as normal (<110 g/m2 in women, <131 g/m2 in men) in 10 patients and increased in 22. Two-dimensional echocardiographic images were obtained with a 5 MHz transducer and digitized off-line. Videodensitometric analysis was performed at end-diastole and end-systole with regions of interest across the septal and posterior wall. The cyclic variation was more pronounced in the left than in the right septal subendocardium (31% +/- 14% vs 16% +/- 14% <0.01) and higher in the subendocardial than in the subepicardial layer of the posterior wall (30% +/- 21% vs 23 +/- 18%, p <0.01). Cyclic variation of the left septal subendocardium was higher in 11 patients with nonhypertrophic ventricles than in 22 with hypertrophic left ventricles (42% +/- 15% vs 27% +/- 12%; p <0.01). The percent cyclic variation of the left septal subendocardium appeared to be much more tightly related to percent systolic thickening in patients with eccentric LV hypertrophy (r=0.80 p <0.01) than in patients with concentric LV hypertrophy (r=0.27, p=0.9) or normal LV mass (r=0.43, p=0.2). A cyclic gray level variation can be consistently detected in different human myocardial regions and layers. It is more obvious in the subendocardial than in the subepicardial layer, and in nonhypertrophic than hypertrophic ventricles. The cyclic subendocardial variation is tightly related to regional systolic thickening in patients with eccentric LV hypertrophy. PMID- 8712146 TI - The heart of the matters of the heart. PMID- 8712147 TI - Clinical use of lossy image compression in digital angiography. PMID- 8712148 TI - Relation of lumen size to restenosis after percutaneous transluminal coronary balloon angioplasty. Lovastatin Restenosis Trial Group. AB - The relation between post-percutaneous transluminal balloon angioplasty (post PTCA) and angiography indexes were investigated in the Lovastatin Restenosis Trial. Post-PTCA percent diameter measured by the operator was found to be a weak predictor of angiographic and clinical follow-up restenosis. PMID- 8712149 TI - An impaired carotid sinus distensibility and baroreceptor sensitivity alter autonomic activity in patients with effort angina associated with significant coronary artery disease. AB - Baroreceptor sensitivity and carotid sinus distensibility were lower in patients with angina associated with significant coronary artery disease than in patients with vasospastic angina. Baroreceptor sensitivity was significantly correlated with carotid sinus distensibility in both groups of patients. PMID- 8712150 TI - Serum electrolytes in children with neurocardiogenic syncope treated with fludrocortisone and salt. AB - We examined the changes in serum electrolytes of patients with syncope treated with salt and fludrocortisone. The most significant change was an increase in serum bicarbonate concentration, with minimal alteration of sodium, potassium, and chloride concentrations; none was clinically significant. PMID- 8712151 TI - Contrast-enhanced doppler ultrasound for noninvasive assessment of pulmonary artery pressure during exercise in patients with chronic congestive heart failure. AB - Pulmonary artery pressure response to exercise was assessed using contrast enhancement of tricuspid regurgitation peak velocities in 19 patients with chronic congestive heart failure. Estimated systolic pulmonary artery pressures correlated closely with invasively measured pressures at rest (r=0.82, p <0.001) and during peak exercise (r=0.86, p <0.001) at a good level of agreement (mean difference 7.3 +/- 12 mm Hg), indicating that this method provides a reliable, noninvasive approach to evaluating functional reserve in patients with chronic congestive heart failure. PMID- 8712152 TI - Transcatheter closure of atrial septal defect or patent foramen ovale with the buttoned device for prevention of recurrence of paradoxic embolism. AB - Transcatheter occlusion of patent foramina ovalia and atrial septal defects in 10 patients with presumptive paradoxic embolic stroke using the buttoned device appears to be encouraging, with safety and efficacy demonstrated during a follow up of up to 60 months. Anticoagulation should be continued until complete disappearance of the right-to-left atrial shunt. PMID- 8712153 TI - Effects of in vivo cocaine administration on human platelet aggregation. AB - To evaluate whether cocaine administration to human volunteers in vivo increases platelet aggregation, 12 healthy male volunteers were studied twice in a prospective, double-blinded fashion. There was a decrease in aggregation following cocaine exposure compared to placebo, which was most prominent at high doses of adenosine diphosphate. PMID- 8712154 TI - Comparison of relation of systolic flow of the right coronary artery to pulmonary artery pressure in patients with and without pulmonary hypertension. AB - Systolic flow velocity is relatively predominant over diastolic velocity in the proximal right coronary artery and right ventricular branch in patients with normal systolic pulmonary artery pressure. The amount of systolic flow velocity reduction in the proximal right coronary artery and right ventricular branch is inversely related to the level of pulmonary hypertension. PMID- 8712155 TI - Short-term hemodynamic effect of a new oral PGI2 analogue, beraprost, in primary and secondary pulmonary hypertension. AB - In 4 patients with primary pulmonary hypertension, there was a -24% +/- 20% decrease in pulmonary vascular resistance, a significant increase of cardiac index by +27 +/- 14% in all 4 patients; a -15 +/- 12% decrease in pulmonary artery pressure in 3 patients; and in 3 patients with 12% secondary pulmonary hypertension, there was a -24 +/- 14% decrease in pulmonary vascular resistance. Beraprost appears to be effective as a new pulmonary vasodilative agent. PMID- 8712156 TI - Does sotalol have reverse-use dependence during tachyarrhythmias? AB - This study assessed the effect of intravenous sotalol on right ventricular effective refractory period at right ventricular pacing rates of 600 and 300 ms cycle length at 3, 6, 9, and approximately 30 minutes after the dose of sotalol. Similar percent increases occurred in the ventricular effective period at the 2 heart rates at all tested times (p >0.2 in each case), and it was concluded that there is no evidence for reverse-use dependence of intravenous sotalol in its effects on right ventricular refractoriness over this range of heart rates. PMID- 8712157 TI - Pericarditis causing exercise test induced ST-elevations. AB - Pericarditis has not been well associated with exercise test induced ST elevations. This case report of a pericarditis patient who underwent exercise stress testing and other similar cases found in the literature suggest that this is an unrecognized clinical manifestation of pericarditis. PMID- 8712158 TI - In memoriam. William S. Kroger, M.D., a notable pioneer in hypnosis and psychosomatic medicine. PMID- 8712159 TI - An interview with William S. Kroger, M.D. Interview by Michael D. Yapko. PMID- 8712160 TI - Multiple susceptibility testing: is it helpful? AB - This study explored whether or not the use of combined group and individually administered susceptibility tests improve the predictive power over the use of a singly administered test. Two hundred and eighty undergraduates were assigned to one of five groups: Group 1 received the HGSHS: A and then the SHSS:C; Group 2 the CIS and SHSS:C; Group 3 the HGSHS: A and the SHCS:A; Group 4 received the CIS and the SHCS:A; and Group 5 was tested on the SHSS:C alone. After the susceptibility screening the subjects were hypnotized and tested on four types of target hypnotic behaviors. From the RSPSHS:I&II the following four factors were chosen (1) cognitive distortion, (2) positive hallucination, (3) negative hallucination, (4) dreams and regression. The items were matched on difficulty level. The data were subjected to a series of stepwise multiple regression and logistic regression analyses. The results confirmed previous research; i.e., (1) The SHSS:C is the best single measure, (2) the SHCS:A is a poor substitute for the SHSS:C; (3) the HGSHS:A is not adequate substitute for SHSS:C; (4) the CIS is weak in predictive power compared to the HGSHS:A; (5) Only for a weak measure such as SHCS:A does combined testing produce an advantage; (6) There appear to be no warm-up effects for SHSS:C when preceded by HGSHS:A. PMID- 8712161 TI - The effects of subject arm position and initial experience on Chevreul pendulum responses. AB - Some clinicians maintain that responses to the Chevreul pendulum illusion are facilitated by resting one's elbow on a table. Others claim the reverse. We compared these two methods in a counterbalanced crossover design by having 32 university students perform the Chevreul pendulum illusion with their elbows supported on a table and with their elbows unsupported. Although there was no main effect for method (elbow supported versus elbow unsupported), subjects who rested their elbows on a table on the first trial were more successful in responding on both trials. This suggests that supporting the elbow does facilitate responding, but only on the initial trial. Performance on subsequent trials is determined by degree of success on the first trial. Similar data from a previous study comparing different hypnotic inductions suggests that this phenomenon is generalizable beyond the Chevreul pendulum illusion and supports the hypothesis that the test-retest reliability of suggestibility scales may be due to a stabilization of response expectancy by a person's first experience of imaginative suggestions. PMID- 8712162 TI - Behavioral and subjective scoring of the Harvard Group Scale of Hypnotic Susceptibility: further data and an extension. AB - The present study replicated and extended Kirsch, Council, and Wickless's work evaluating the reliability and concurrent validity of a subjective response scale developed for the Harvard Group Scale of Hypnotic Susceptibility. Consistent with Kirsch et al.'s findings, the subjective counterpart of the Harvard scale demonstrated high reliability (coefficient .90) and concurrent validity (r = .84 with Harvard's standard behavioral score). Similar correlations of the behavioral and subjective scales with the Dissociative Experience Scale (DES) and Phenomenology of Consciousness Inventory (PCI) suggested that the two hypnotizability scales are measuring similar constructs. However, using the two scales in combination did not yield a morestringent measure of hypnotizability in terms of DES and PCI variables than using either of them alone. PMID- 8712163 TI - Hypnosis in the treatment of patients with severe burns. AB - Burn injuries are a frequent form of trauma, the care for which typically involves repeated, intrusive procedures and acute, excruciating levels of pain. Although research in the use of hypnosis with burn patients is largely anecdotal there is emerging evidence that the burn unit may be one of the most useful arenas for the clinical application of this technique. The acute, identifiable nature of burn care procedures and the emotional state of patients in trauma care both provide an often receptive setting for the use of this intervention. PMID- 8712164 TI - The impact of a confusion technique on hypnotic responsivity in low-susceptible subjects. AB - The present study investigated the hypothesis that a confusion technique would produce greater hypnotic responsiveness for subjects who responded minimally to a standard hypnotic induction. Ten subjects who scored in the low range of hypnotic susceptibility on the HGSHS:A (scores of 0-4) participated in a confusion induction session. Hypnotic responsiveness was measured by several scales: the Harvard Group Scale of Hypnotic Susceptibility (HGSHS:A), a Depth Estimate (DE), and the Inventory Scale of Hypnotic Depth (ISHD). Qualitative data was collected via a follow-up interview. Collectively, the results did not support the hypothesis that a confusion technique is more effective than a standard induction for low-susceptible subjects. Implications for future research will be discussed. PMID- 8712165 TI - Hypnosis training in psychology intern programs. AB - The use of hypnosis is becoming increasingly common in clinical practice. This article examines hypnosis training in psychology internship programs. A survey was sent to all APA intern programs listed in the APPIC directory requesting information about opportunities for training and utilizing hypnosis during the intern year. Thirteen percent of responding programs (n = 424) reported regular seminars in hypnosis, while another 37% offer elective opportunities. However, of those programs that provide training, the average amount of time was approximately seven hours. The authors conclude that while training is provided to a significant percentage of interns, the amount of time is not enough to allow for its ethical use. It is suggested that as the use of hypnosis increases, training programs may want to consider opportunities for more adequate hypnosis training. PMID- 8712166 TI - Testing for heredity hypercoagulability Activated protein C resistance. PMID- 8712167 TI - Prevention of transfusion-associated cytomegalovirus infection. Practice parameter. American Society of Clinical Pathologists. AB - This practice guideline represents the opinions and recommendations of the author(s), the American Society of Clinical Pathologists (ASCP) Practice Parameters Committee and the ASCP Board of Directors regarding the appropriate strategies for each clinical condition or laboratory test discussed in this guideline. This guideline is designed primarily as an educational resource for physicians in the provision of quality medical services. Adherence to this guideline is completely voluntary and does not necessarily assure a successful medical treatment or result. This practice guideline should not be considered inclusive of all proper procedures and tests or exclusive of other procedures or tests that are reasonably directed to obtaining the same results. The physician should apply his or her own professional judgment to the unique clinical circumstances presented by the particular procedure or test. Physicians are encouraged to document the reasons for whatever procedure or test they use (whether or not in conformance with this guideline). Physicians should also take care to consider other medical and scientific advances that are available after the date of adoption of this guideline. This practice guideline was developed exclusively for the purposes set forth above and not for use in connection with matters involving reimbursement, credentialing, or utilization review. PMID- 8712168 TI - Rubella immunity. Defining the level of protective antibody. AB - The Rubella Subcommittee of the National Committee for Clinical Laboratory Standards has proposed lowering the breakpoint to define rubella immunity from 15 to 10 IU/mL. This recommendation stems from epidemiologic studies on vaccinated persons with low levels of antibody and anecdotal reports. Additional support comes from Centers for Disease Control and Prevention studies and reports. The effectiveness of rubella vaccination is well documented and the 10 IU/mL antibody level is protective in the vast majority of persons. Sporadic reports of viremia and/or reinfection among previously immunized persons with low antibody levels have been reported but proven cases of reinfection have also occurred in persons with titers greater than or equal to the 15 IU/mL cut-off. Despite the occasional occurrence of rubella reinfection in persons with low titers, the theoretical risks are small especially as compared with significantly greater risk in persons who have not been vaccinated. Immunity in a given patient is a clinical decision and the results of antibody tests for rubella, like other laboratory tests, must be evaluated in the context of the clinical setting. PMID- 8712169 TI - Use of a single slide trichrome-stained concentrate for the detection of intestinal parasites. Stained concentration procedure for ova and parasites. AB - The detection and identification of intestinal parasites was studied using a single slide of a trichrome-stained concentrate made from a cupric-polyvinyl alcohol (PVA)-preserved specimen. When saline was used in place of formalin in the concentration procedure, it was possible to adequately stain the concentrate with a modified-trichrome stain. This was compared to a more traditional two slide ova and parasite (O&P) procedure. Of the 1,000 specimens evaluated, a total of 261 parasites were detected using the single slide, modified trichrome-stained concentrate compared to 249 from the formalin/Hemo-De wet mount and 226 from the unconcentrated trichrome stain. The detection rate of the single slide method was comparable to the combined detection rate of the two-slide method. The saline/Hemo-De concentration procedure has the advantage of using nontoxic saline in place of formaldehyde. This new method is more cost-effective because it requires the processing and evaluation of only one slide. PMID- 8712170 TI - Use of viral cultures and serologic tests for cytomegalovirus infection. Rational or random? AB - To assess their applied clinical utility, viral cultures and serological tests for cytomegalovirus (CMV) were reviewed at Duke University Medical Center (DUMC), a 1,125-bed tertiary-care hospital. Less than 1% (3 of 1,216) of CMV cultures were positive, and 8% of serum samples (45 of 587) were positive by single sera IgM ELISA. Sixteen percent (32 of 199) of IgG acute to convalescent sera pairs were positive. Four hundred five of 588 (69%) serum samples were positive for the IgM/IgG passive latex agglutination test, consistent with the results for random blood donors. Review of hospital records showed that fewer than 1% of the positive-test patients (excluding the latex test) received treatment for CMV. Comparisons of tests ordered on individual patients did not disclose a coherent diagnostic strategy. The authors conclude that the majority of testing for CMV in their medical center does not yield useful clinical information, but carries a substantial financial burden. A new diagnostic strategy to attempt to diagnose CMV disease is needed. PMID- 8712171 TI - Lymphoid-associated antigen expression by acute myeloid leukemia. AB - The expression of lymphoid-associated antigens (LAA) on blasts in acute myeloid leukemia (AML) and myeloproliferative disorders in myeloid blast crisis (MPD/MBC) has often been used to establish a diagnosis of acute mixed lineage leukemia (AMLL). The purpose of this study was to determine the incidence of LAA expression in AML and MPD/MBC (Ly + AML); to assess lymphoid differentiation at the genomic level in Ly + AML; and to compare features of Ly + AML with AML and MPD/MBC lacking these antigens (Ly-AML). Seventy-four consecutive cases of AML and MPD/MBC were reviewed for blast morphology, TdT reactivity, and cytochemistry results. Blast immunophenotyping was performed by multiparameter flow cytometry. Acute myeloid leukemia was subtyped according to the FAB classification. Acute myeloid leukemia and MPD/MBC cases expressing one or more of the following antigens, CD2, CD3, CD5, CD7, CD19, or CD20, were considered to be Ly + AML. Immunoglobulin (Ig) and T-cell receptor (TCR) gene rearrangement studies were performed by Southern blot analysis using probes for JH, Jkappa, and JBI/BII. Sixteen of the 74 cases (22%) were identified as Ly + AML. Of these, the T-cell associated markers CD7, CD2, and CD5 were expressed on 7(44%), 6(38%), and 4(25%) Ly + AML cases, respectively. The B-cell-associated markers CD19 and CD20 were expressed on two cases (13%) and one (6%) case, respectively. The FAB subtypes were similarly represented among Ly + AML and Ly-AML. Expression of LAA did not correlate with TdT positivity. In nine cases of Ly + AML (7 expressing T-cell associated antigens and two expressing B-cell-associated antigens), Southern blot analysis revealed no Ig or TCR gene rearrangements. These results suggest that expression of CD2, CD5, and CD7 in otherwise straightforward AML should not be taken as evidence of lymphoid lineage commitment and does not warrant a diagnosis of AMLL. PMID- 8712172 TI - CD117/CD34 expression in leukemic blasts. AB - CD117 is a transmembrane protein receptor encoded by the c-kit proto-oncogene. The CD117 ligand is stem cell factor, an important hematopoietic regulator. CD117 is present on approximately 4% of normal bone marrow mononuclear cells and in acute myelogenous leukemia (AML) and chronic myelogenous leukemia in myeloid blast crisis, but rarely in acute lymphoblastic leukemia (ALL). Initially viewed as a primitive myeloid marker, CD117 has been identified in all FAB subtypes of AML and may predict poor outcome. CD34, a primitive stem cell marker, may also predict poor outcome. The aim of this study was to examine the relationship between CD117 and CD34 expression on leukemic blasts and to determine whether CD117 is related to lymphoid-associated antigen (LAA) expression in AML. Consecutive bone marrow samples were studied from cases of AML (30 cases), myelodysplastic syndromes (MDS) (4 cases), myeloproliferative disorders in blast crisis (MPD-BC) (6 cases), and ALL (5 cases). Cases were diagnosed according to FAB criteria and included M0 (3 cases), M1 (2 cases), M2 (13 cases), M3 (1 case), M4 (6 cases), M5 (3 cases), M6 (1 case), AML NOS (1 case), RAEB (3 cases), and RAEB-T (1 case). CD117 and CD34 were analyzed by multiparameter flow cytometry. Blasts in 10 de novo AML samples were CD117+/CD34+ in 4 cases, CD117+/CD34-in 3 cases, CD117-/CD34+ in 1 case, and CD117-/ CD34- in 2 cases. Blasts in 20 cases of relapsed AML were CD117+/ CD34+ in 13 cases, CD117+/CD34- in 6 cases, and CD117-/CD34+ in 1 case. Blasts in MDS were CD117+/CD34+ in 3 cases, CD117-/ CD34+ in 1 case. Blasts in MPD-BC were CD117+/CD34+ in 4 cases, CD117-/CD34+ in 2 cases. Blasts in ALL were CD117+/CD34+ in 1 case, CD117-/CD34+ in 1 case, CD117 /CD34- in 3 cases. Of 26 cases of CD117+ AML, CD4 was expressed in 15 (58%) cases, CD7 in 7 (27%) cases, and CD2 in 2 (8%) cases. CD117/CD34 expression did not correlate with FAB subtype of AML. CD117 is borne on most leukemic blasts of myeloid origin (in this study, 87% of AML, 80% of MPD-myeloid BC, and 75% of MDS) and does not exclude expression of LAA. Although CD117 is a receptor for stem cell factor, its expression does not appear to correlate with CD34 positivity. PMID- 8712173 TI - Mantle cell lymphoma. Morphologic findings in bone marrow involvement. AB - Although mantle cell lymphoma (MCL), has been well described in lymph nodes, involvement of blood and bone marrow has not been well defined. The authors reviewed involved blood and marrow specimens from 13 patients with MCL to determine patterns of infiltration. These findings were compared to marrow involvement by follicular small cleaved cell lymphoma (SCCL) and small lymphocytic lymphoma (SLL). Peripheral blood involvement by MCL was present in 5 patients (38%). The circulating lymphoma cells were small (7-10 mu) with slightly folded nuclei. Marrow involvement ranged from 5% to 90% of the marrow space and was predominantly intertrabecular, including nodules and interstitial infiltrates (9 cases each; 68%). Paratrabecular aggregates (6 cases; 46%) and diffuse replacement by lymphoma (3 cases; 23%) were also seen. In SCCL, paratrabecular involvement was seen as were interstitial nodules. Cases of SLL showed diffuse, interstitial or nodular involvement without paratrabecular localization. Cytologic comparison showed nuclei that were angulated in SCCL, round in SLL, and slightly irregular in MCL, with considerable overlap among the groups. The architectural and cytologic findings in marrow involved by MCL show features of both SCCL and SLL, and cannot be used to definitively diagnose MCL. PMID- 8712174 TI - The evaluation of various mathematical RBC indices and their efficacy in discriminating between thalassemic and non-thalassemic microcytosis. AB - The differentiation between thalassemic and non-thalassemic microcytosis has important clinical implications in hematology and medicine. A simplified index, based on red cell parameters derived from automated blood cell analyzers, which could be used to discriminate between microcytic patients with a high probability of thalassemia minor and those with a low probability, would be an extremely useful tool. Five mathematical indices have been proposed as useful for this purpose. These are the: Bessman index, Shine and Lal index, England index, Mentzler index, and mean cell volume (MCV) alone. This study was designed to prospectively evaluate the efficacy of these indices. Patient samples were chosen every fourth day from all patient samples referred to the hematology laboratory at St. Joseph's Hospital over a 6-month period. All patient samples with an MCV < 80 fL and age > or = 18 years were considered eligible for the study. After enrollment and laboratory analysis were complete sensitivities and specificities were calculated for each of the indices using a variety of cut-off values and receiver operator characteristic (ROC) curves were constructed. Based on statistical analysis of the area under these curves, the authors conclude that MCV alone is as effective as the Mentzler and Shine and Lal indices in selecting microcytic patient samples with a high probability of thalassemia minor for thalassemia testing. They also conclude that the Bessman index and the England index are ineffective indices for this purpose. PMID- 8712175 TI - Acute lymphoblastic leukemia in a case of essential thrombocythemia. AB - Essential thrombocythemia is a myeloproliferative disorder that infrequently evolves into acute leukemia. Leukemic transformation is frequently preceded by therapy with alkylating agents or radioactive phosphorus (32P), and is virtually always myeloid in nature. In this report, the authors describe a case of acute lymphoblastic leukemia arising in a patient with long-standing essential thrombocythemia. PMID- 8712176 TI - Lymph node recovery from colorectal resection specimens removed for adenocarcinoma. Trends over time and a recommendation for a minimum number of lymph nodes to be recovered. AB - Recovery of pericolorectal lymph nodes from colectomy specimens has long been part of colorectal cancer staging. Recently, adjuvant therapy has been added for high stage carcinomas, providing further impetus for performing careful lymph node dissections. Pericolorectal lymph nodes were examined to determine if there has been a change over time in the number of lymph nodes recovered and proportion of specimens with pericolonic lymph node metastases from colorectal carcinoma resection specimens. Also, the authors attempted to establish a recommendation for a minimum number of lymph nodes that should be recovered before a colon can be considered free of metastases. Slides and reports of the first 20 consecutive pT3 colorectal carcinoma resections in each year from 1955 to 1995 at William Beaumont Hospital that did not have known metastases at the time of surgery were reviewed (750 specimens total). The mean number of lymph nodes recovered per specimen and incidence of detected lymph node metastases increased over the 41 year period, with the greatest increase occurring during 1992-1995. The greatest proportion of patients with lymph node metastases detected occurred in the 17 to 20 lymph nodes recovered per specimen group. Specimens with more than 20 lymph nodes did not have a higher proportion of lymph node metastases detected compared to specimens with 17 to 20 lymph nodes. Approximately 20% of the specimens with metastases had more than 17 lymph nodes recovered. These results suggest that pathologists should retrieve all the lymph nodes that can be recovered, but at least 17 lymph nodes should be recovered to insure accurate documentation of nodal metastases when present. PMID- 8712177 TI - Solitary fibrous tumor of the meninges: a lesion distinct from fibrous meningioma. A clinicopathologic and immunohistochemical study. AB - Seven solitary fibrous tumors (SFTs) of the meninges are presented and their clinicopathologic features are compared with those of 64 fibrous meningiomas (FM). Patients with SFT included 5 females and 2 males age 47 to 73 years. The dura-based tumors involved the parasagittal region (1), tentorium (2), cerebellopontine angle (2), and spinal region (2). One each showed invasion of brain and of a spinal nerve root. Of four SFTs with at least 1-year follow-up, one subtotally resected example recurred. No tumors metastasized. All consisted of spindle cells disposed in fascicles between prominent, eosinophilic bands of collagen. Whorls and storiform cell arrangements were lacking. Mitoses ranged from 1 to 7/10 400 x fields. MIB-1 labeling indices ranged from 1% to 18% (mean 4%). All were PAS negative and showed strong immunoreactivity for vimentin and CD34. Of cases studied, half were estrogen and all were progesterone receptor immunopositive. The majority (72%) of FMs occurred in females and most (72%) were supratentorial. Recurrence was noted in 15%. Mitotic activity varied from 0 to 3 mitoses per 10 400 x fields (mean < 1). MIB-1 labeling indices ranged from 1% to 5% (mean 1.5%). Unlike SFT, FMs were glycogen-containing and variously exhibited a storiform pattern (13 of 20), psammoma body formation (9 of 20), and calcification of collagen (4 of 20). Immunoreactivities included vimentin (100%), focal to patchy EMA (80%), S-100 protein (80%), collagen IV (25%), and patchy, mild-to-moderate CD34 staining (60%). Of cases studied, nearly half were estrogen and all were progesterone receptor staining positive. Meningeal SFTs represent a distinct morphologic entity, the morphologic and immunohistochemical features of which differ from those of FM and suggest a histogenetic relationship to pleural SFT. Although a minority histologically appear to be low grade malignant, our limited experience suggests that they behave in a benign fashion. The classification of mesenchymal tumors affecting the central nervous system must be expanded to include SFT. PMID- 8712178 TI - To freeze or not to freeze. A comparison of methods for the handling of breast biopsies with no palpable abnormality. AB - Recently there have been several editorials suggesting that breast biopsies with small or nonpalpable lesions should not be subjected to frozen section examination. In this study, the approach of routinely performing frozen sections on all breast biopsies was compared to the recommended approach of performing frozen sections only on cases with gross lesions < 1.0 cm. From July 1, 1992 through June 30, 1993, there were 444 breast biopsies submitted. Nearly all of these (98%) had frozen sections performed. Among these cases, there were 14 false negative frozen section diagnoses equating to a frozen section sensitivity of 84%, and a frozen section false-negative rate of 3.3%. From July 1, 1994 through June 30, 1995, there were 601 breast biopsies submitted. Of these cases, 310 contained a gross lesion > 1.0 cm and were subjected to frozen section examination. The remaining 291 cases contained either no distinct lesion or a gross lesion < 1.0 cm. These cases were not frozen. Among the 310 cases that were frozen, there were 3 false-negative frozen section diagnoses equating to a frozen section sensitivity of 96%, and a frozen section false-negative rate of 1.0%. These findings indicate that careful gross examination combined with selective freezing of only cases containing a gross lesion > 1.0 cm improves frozen section accuracy. This study provides statistical support for previous recommendations that frozen section examination should be limited to cases with distinct gross lesions > 1.0 cm. PMID- 8712179 TI - Sialolithiasis. Differential diagnostic problems in fine-needle aspiration cytology. AB - Sialolithiasis with obstruction of major salivary gland ducts can lead to clinical tumefaction related to cystic dilatation. In addition to mucus accumulation, these pseudoneoplasms feature hyperplasia and squamous metaplasia of the ductal lining epithelium, with varying degrees of inflammation. The authors report five examples of this lesion aspirated from two males and three females ranging in age from 45 to 80 years (median 65 years). Three were in the submaxillary gland, and two were in the parotid. In three cases, stone fragments were identified, and diagnoses of sialolithiasis were rendered; two of these patients underwent surgical excision. The remaining two cases showed prominent foam cells and metaplastic squamous cells in a mucoid background that mimicked low grade mucoepidermoid carcinoma. Stone fragments were not identified and a differential diagnoses of sialolithiasis versus low grade mucoepidermoid carcinoma were suggested. Surgical excision revealed sialolithiasis in both instances. When stone fragments are identified in aspirated material, these cases pose little diagnostic difficulty. However, when this material is not present, epithelial changes and mucus accumulation may be difficult to distinguish from low grade mucoepidermoid carcinoma. Cautious interpretation is suggested in this setting. PMID- 8712180 TI - Aberrations of chromosomes 9 and 17 in bilharzial bladder cancer as detected by fluorescence in situ hybridization. AB - Fluorescence in situ hybridization (FISH), using repetitive alpha satellite probes to chromosomes 9 and 17, was performed on 27 paraffinembedded bladder specimens from 18 Egyptian patients affected with bilharzial carcinoma. Fluorescence in situ hybridization results in the carcinomas and benign mucosa of patients with schistosomiasis were compared with flow cytometric DNA ploidy and cell cycle assays. These results demonstrate the feasibility of this technique for studies of these archival specimens, and suggest early onset of chromosomal aberrations in histologically benign mucosa of schistosomal cystitis with progression during the development of bladder cancer. On the basis of these data, monosomy 9 may be an early chromosomal change in urothelium of the bilharzia infested bladder, and a predictor of incipient carcinoma in patients with bilharzial cystitis. PMID- 8712181 TI - Salivary duct carcinoma secreting prostate-specific antigen. AB - Prostate-specific antigen (PSA) is a 30 kDa glycoprotein serine protease that shows high tissue specificity for prostatic tissue, both benign and malignant. However, recent reports have shown that a variety of normal and neoplastic tissue types express PSA immunohistochemically. In addition, rare instances of the secretion of PSA by nonprostatic cancers have been reported in the literature. The authors present a case of salivary duct carcinoma associated with elevated serum levels of PSA. Both the primary tumor and metastases stained positively with anti-PSA monoclonal antibodies, but were negative with antibodies directed against prostate-specific acid phosphatase. Elevated serum PSA levels were confirmed with three different immunoassay methods. A peak serum level of 140 micrograms/L was measured and this correlates with levels of PSA associated with metastatic prostatic carcinoma. High performance liquid chromatography with a molecular sieve column characterized the serum PSA into both free protein (approximately 20%) and protein bound to alpha-1-antichymotrypsin (PSA ACT)(approximately 80%). Molecular weights of the free PSA and PSA-ACT subfractions were 27-31 kDa and 100-110 kDa, respectively. PMID- 8712182 TI - Utilization of testing for activated protein C resistance in a reference laboratory. AB - Activated protein C (APC) resistance is a newly described thrombotic disorder accounting for the majority of patients with inherited thrombosis. The authors prospectively evaluated laboratory testing of this disorder over a 1-year period in their reference laboratory, which draws samples from a large number of community and academic hospitals throughout the United States. Testing for other inherited thrombotic disorders (antithrombin III deficiency, protein C and S deficiencies) occurred at a six-fold greater rate than that for APC resistance. Previously published studies have indicated a prevalence of up to 60% for APC resistance in populations with thrombosis; however, the prevalence rate in this study was only 12%. Of patient samples submitted for APC resistance assays, 37% were not evaluable because of concomitant anticoagulant therapy. Clinical pathologists and practitioners need to be made aware of APC resistance and of optimal sample collection to improve the efficiency of laboratory testing for thrombosis. PMID- 8712183 TI - Evaluation of the APEC analyzer. PMID- 8712184 TI - Parents and peer group as mediators of the effect of community structure on adolescent problem behavior. AB - Used a sample of 207 single-parent families residing in 104 small, Midwestern communities to test hypotheses regarding the link between community context and adolescent conduct problems and psychological distress. For boys, community disadvantage had a direct affect on psychological distress, while it indirectly boosted the probability of conduct problems by disrupting parenting and increasing affiliation with deviant peers. Community disadvantage was unrelated to the deviant behavior or emotional well-being of girls. Proportion of single parent households in the community had a direct effect on girls' conduct problems. It also contributed indirectly to girls' conduct problems by increasing the probability of involvement with deviant peers. Possible explanations for these gender differences are provided. PMID- 8712185 TI - Physical characteristics of residential psychiatric and substance abuse programs: organizational determinants and patients outcomes. AB - Described the Physical and Architectural Characteristics Inventory (PACI), which assesses the physical characteristics of hospital- and community-based psychiatric and substance abuse residential treatment programs for adults. Developed using data from 94 representative programs, the PACI has 7 dimensions that assess community accessibility, physical features that add convenience, aid recreation, and provide support for patients, and space for patient and staff functions. In comparison to community-based programs (n = 63), hospital programs (n = 31) had more social-recreational and prosthetic aids, safety features, staff facilities, and space. The PACI dimensions were also related to other organizational features, such as facility ownership and program size and staffing level. Programs with more physical amenities had patients who were more involved in self-initiated and community activities and were more likely to successfully complete the program and be discharged to independent living situations and paid jobs. Programs with more social-recreational and prosthetic aids, space, and access to community resources also had better patient outcomes. In addition, PACI dimensions were related to observers' ratings of facility attractiveness. Potential applications of the PACI and the full inventory of which it is a part are discussed. PMID- 8712186 TI - Understanding environments: the key to improving social processes and program outcomes. AB - The contributions to this Special Issue illustrate research on several important types of environmental units, including psychiatric and substance abuse treatment programs, neighborhood block groups, and entire communities. They also exemplify alternative methodologies, such as assessing environments by relying on participants' appraisals, external observers' ratings, historical archives, and direct observation. I draw on these contributions and some of my own work to discuss four recurrent issues: (a) how to conceptualize environmental domains and dimensions; (b) how to understand environmental dynamics, that is, the interplay of environmental factors both within one setting and across settings; (c) how to comprehend the processes that link environmental factors to outcomes, especially with respect to the power and evanescence of environmental influence; and (d) how to use information about environments to monitor and improve them. Increased knowledge in these four areas is essential to help fulfill a major aim of community psychology: to understand social processes and improve intervention program outcomes. PMID- 8712187 TI - An ecological assessment of community-based interventions for prevention and health promotion: approaches to measuring community coalitions. AB - Presented an ecological assessment of a community coalition to prevent alcohol, tobacco, and other drug abuse, and related risks. Ecological assessment is defined as occurring at multiple social levels and along a continuum of stages of coalition readiness. The assessment is aided by the triangulation, or combining of assessment methods and strategies. Measures used to assess the coalition's formation, implementation of community initiatives, and production of community impacts are described, along with the triangulation strategies used to enhance the assessment findings. PMID- 8712188 TI - Ecological assessments of community disorder: their relationship to fear of crime and theoretical implications. AB - Researchers suggest that fear of crime arises from community disorder, cues in the social and physical environment that are distinct from crime itself. Three ecological methods of measuring community disorder are presented: resident perceptions reported in surveys and on-site observations by trained raters, both aggregated to the street block level, and content analysis of crime- and disorder related newspaper articles aggregated to the neighboring level. Each method demonstrated adequate reliability and roughly equal ability to predict subsequent fear of crime among 412 residents of 50 blocks in 50 neighborhoods in Baltimore, MD. Pearson and partial correlations (controlling for sex, race, age, and victimization) were calculated at multiple levels of analysis: individual, individual deviation from block, and community (block/neighborhood). Hierarchical linear models provided comparable results under more stringent conditions. Results linking different measures of disorder with fear, and individual and aggregated demographics with fear inform theories about fear of crime and extend research on the impact of community social and physical disorder. Implications for ecological assessment of community social and physical environments are discussed. PMID- 8712189 TI - Relation of smoking and alcohol consumption to serum fatty acids. AB - To examine the relation of cigarette smoking and alcohol consumption to serum fatty acid levels, the authors conducted a cross-sectional study of 190 men who were enrolled in the Multiple Risk Factor Intervention Trial between 1973 and 1976. After controlling for dietary fat, cholesterol, energy intake, and other potential confounders, the authors found that smoking and alcohol intake were associated with the serum cholesterol ester and phospholipid levels of several fatty acids. As the number of cigarettes smoked per day increased, the levels of cholesterol ester and phospholipid palmitoleic acid (16:1) and oleic acid (18:1) and the levels of phospholipid dihomogammalinolenic acid (20:3) and omega-9 eicosatrienoic acid (20:3) increased (all p's < or = 0.01). Serum levels of phospholipid omega-3 docosahexaenoic acid (22:6) and cholesterol ester and phospholipid arachidonic acid (20:4) were inversely associated with smoking (all p's < or = 0.01). As the number of alcoholic drinks per week increased, levels of cholesterol ester and phospholipid palmitic acid (16:0) and oleic acid (18:1), cholesterol ester myristic acid (14:0), and phospholipid palmitoleic acid (16:1), adrenic acid (22:4), and omega-9 eicosatrienoic acid (20:3) increased (all p's < 0.05), whereas levels of cholesterol ester and phospholipid linoleic acid (18:2) and phospholipid stearic acid (18:0) and the serum polyunsaturated fat: saturated fat ratio decreased (all p's < or = 0.01). These results suggest that smoking and alcohol consumption may influence the absorption, synthesis, or metabolism of serum fatty acids. Studies that use serum fatty acid levels as indicators of dietary fat intake should control for the effects of cigarette smoking and alcohol consumption. PMID- 8712190 TI - Sex-specific associations of magnetic resonance imaging-derived intra-abdominal and subcutaneous fat areas with conventional anthropometric indices. The Atherosclerosis Risk in Communities Study. AB - Accurate measurement of central fat patterning is difficult to obtain by conventional anthropometry. Direct measurement of intra-abdominal fat area by magnetic resonance imaging, while accurate, is impractical for large-scale observational studies. This report examines the sex-specific associations of conventional anthropometric indices with intra-abdominal fat and subcutaneous fat areas measured by magnetic resonance imaging. A total of 157 volunteers (97 men and 60 women) aged 48-68 years of predominately white ethnicity had intra abdominal fat and subcutaneous fat areas measured as part of the Atherosclerosis Risk in Communities (ARIC) Study. Weight, body mass index, waist circumference, waist : hip ratio, and subscapular skinfold thickness were measured or calculated by a standardized protocol. On average, women had a lower intra-abdominal fat area than men (109.5 cm2 vs. 152.9 cm2) but a higher mean subcutaneous fat area (287.8 cm2 vs. 214.6 cm2). After adjustment for age, intra-abdominal fat area was quadratically associated with body mass index, waist circumference, weight, and subscapular skinfold thickness in men; in women, these associations were best modeled by a positive linear equation. Waist : hip ratio was linearly related to intra-abdominal fat area in both sexes. In general, anthropometric measures predicted lower percentages of the total variance in intra-abdominal fat area for men than for women. For subcutaneous fat area, all anthropometric indices were linearly associated and predicted more of the variance in subcutaneous fat area than in intra-abdominal fat area. These results indicate that among men, greater intra-abdominal fat deposition rates occur at relatively low body weights and fat is more uniformly deposited at higher weights. Women appear to deposit intra abdominal fat at a constant rate as they gain weight, even after menopause. The authors conclude that when waist circumference or body mass index is used as a surrogate for intra-abdominal fat area in men, a quadratic term should be included in the analysis as a predictor variable. Subcutaneous fat area can be estimated well by linear measures commonly employed in epidemiologic studies. PMID- 8712191 TI - Alcohol intake and subjective health. AB - This cross-sectional analysis examined associations between alcohol intake and subjective health in a random sample (n = 6,040) drawn from the general population aged 25-64 years in Finland in 1992. Self-reported health was good for 3,375 persons and average or poor (suboptimal) for 2,665 persons. Crude odds ratios suggested a U-shaped pattern between alcohol intake and suboptimal health. The pattern took more of a J-shape after data were controlled for sex, age, education, marital status, lack of close friends, being on a disability pension, smoking, being an ex-drinker, and having decreased one's alcohol intake during the past 12 months because of health problems. An interaction was found between alcohol and smoking. The pattern of alcohol odds ratios showed a J-shaped association among never smokers, and a similar pattern was suggested among ex smokers and current smokers. Among never smokers, the lowest risk was found at the alcohol consumption level of 100-199 g/week (odds ratio (OR) = 0.58, 95% confidence interval (CI) 0.38-0.89). The highest risk was found among persons who regularly smoked > or = 20 cigarettes per day and drank > or = 300 g/week (OR = 4.44, 95% CI 2.36-8.36). The risk for ex-drinkers did not differ from that for lifelong abstainers (OR = 0.89, 95% CI 0.62-1.28), but persons who had decreased their alcohol intake during the past 12 months because of health problems had a higher risk (OR = 1.21, 95% CI 1.05-1.39). The authors conclude that moderate alcohol intake is related to a self-perception of good health. PMID- 8712192 TI - Community education for cardiovascular disease prevention. Morbidity and mortality results from the Minnesota Heart Health Program. AB - The Minnesota Heart Health Program was a community trial of cardiovascular disease prevention methods that was conducted from 1980 to 1990 in three Upper Midwestern communities with three matched comparison communities. A 5- to 6-year intervention program used community-wide and individual health education in an attempt to decrease population risk. A major hypothesis was that the incidence of validated fatal and nonfatal coronary heart disease and stroke in 30- to 74-year old men and women would decline differentially in the education communities after the health promotion program was introduced. This hypothesis was investigated using mixed-model regression. The intervention effect was modeled as a series of annual departures from a linear secular trend after a 2-year lag from the start of the intervention program. In the education communities, 2,394 cases of coronary heart disease and 818 cases of stroke occurred, with 2,526 and 739 cases, respectively, being seen in the comparison communities. The overall decline in coronary heart disease incidence was 1.8 percent per year in men (p = 0.03) and 3.6 percent per year in women (p = 0.007). For stroke, there were no significant secular trends. The authors recently published findings showing minimal effects of sustained intervention on risk factor levels. In the current report, there was no evidence of a significant intervention effect on morbidity or mortality, either for coronary heart disease or for stroke. PMID- 8712193 TI - Differences in risk factors for epithelial ovarian cancer by histologic type. Results of a case-control study. AB - A case-control study of associations between dietary and reproductive factors and cancer of the ovary was conducted during 1989-1992 in metropolitan Toronto and nearby areas of southern Ontario, Canada. In total, 450 women aged 35-79 years with histologically verified new primary epithelial ovarian cancers were interviewed concerning their reproductive history and dietary practices. Over the same time period, 564 randomly selected population controls, frequency-matched to the cases according to three 15-year age groups, were also interviewed. Continuous unconditional logistic regression methods were used for analysis. It was found that childbearing and use of oral contraceptives were associated with significant decreasing trends in the risk of epithelial ovarian cancer of all principal histologic types except mucinous tumors. For each full-term pregnancy, the odds ratio was 0.76 (95% confidence interval (CI) 0.69-0.85) for nonmucinous tumors and 1.03 (95% CI 0.88-1.21) for mucinous tumors; for each year of oral contraceptive use, the odds ratio was 0.89 (95% CI 0.85-0.93) for nonmucinous tumors and 0.98 (95% CI 0.93-1.04) for mucinous tumors (p = 0.00051 and p = 0.0040, respectively, for the difference in odds ratios between mucinous and nonmucinous tumors). Saturated fat intake also appeared to convey greater increased risk for women with mucinous tumors than for women with neoplasms of other histologic types (p = 0.029). Among women with nonmucinous tumors, increasing trends in risk of invasive serous cancer (p = 0.018), and particularly endometrioid cancer (p = 0.0041), were seen with use of noncontraceptive estrogens. Otherwise, borderline-malignant neoplasms seemed to have a similar spectrum of risk factor associations as invasive cancers. On the basis of this study and a number of others, the authors suggest that mucinous ovarian tumors may be etiologically unrelated to other types of epithelial tumors, and thus should be considered separately in studies of ovarian cancer. PMID- 8712194 TI - Risk of breast cancer among white women following induced abortion. AB - Some studies (but not all) have suggested that there may be an increase in the risk of breast cancer associated with a prior induced abortion. The risk, if present, may vary according to the duration of the pregnancy in which the abortion occurred, or to a woman's age or parity at that time. The authors conducted a case-control study of breast cancer in white women under age 45 years to address the question of breast cancer risk in relation to induced abortion, with the intention of identifying subgroups of women who might be at particularly increased risk. White women who were diagnosed with breast cancer (n = 1,302) from May 1, 1990, through December 31, 1992, in three geographic regions of the United States (Atlanta, Georgia; Seattle/Puget Sound, Washington; and five counties in central New Jersey) were interviewed about their reproductive histories, including the occurrence of induced abortion. Similar information was obtained from control women identified through random digit dialing. Logistic regression analysis was used to estimate the relative risk of breast cancer associated with a history of induced abortion, controlling for the potentially confounding influence of other breast cancer risk factors. Among women who had been pregnant at least once, the risk of breast cancer in those with a prior induced abortion was 20% higher than that in women with no history of abortion (95% confidence interval 1.0-1.5). This small increase in risk varied little according to number of abortions or a woman's current age. The association was present primarily among nulliparous women whose abortions occurred prior to 9 weeks' gestation (estimated relative risk = 2.0, 95% confidence interval 1.2 3.3). There was no excess risk of breast cancer associated with induced abortion among parous women. These data support the hypothesis that there may be a small increase in the risk of breast cancer related to a history of induced abortion among young women of reproductive age. However, the data from this study and others do not permit a causal interpretation at this time; neither do the collective results of the studies suggest that there is a subgroup of women in whom the relative risk associated with induced abortion is unusually high. PMID- 8712195 TI - Predictors of ovarian steroid secretion in reproductive-age women. AB - During the baseline period (1985-1988) of a prospective study, midcycle and luteal-phase estrogens and progestins were measured in 175 healthy women aged 21 36 years with spontaneous, cyclic menses in Brooklyn, New York. Subjects contributed daily first-morning urine specimens and three blood specimens during a single menstrual cycle monitored by basal body temperature. Hormone levels were compared according to age, race, and levels of known or suspected breast cancer risk factors. Late age at menarche was associated with increased urinary and serum progestin levels. Increased body weight was associated with decreased progestin levels, even in ovulatory women. Neither weight nor age at menarche was related to estrogen levels. Cigarette smoking was associated with decreased midcycle and luteal-phase estradiol levels. No other factors were associated with differences in any of the hormones measured either midcycle or during the luteal phase, despite good statistical power to detect moderate differences. Sources of individual variability in ovarian steroid levels remain unexplained. These data do not support hypotheses that breast cancer risk factors act through an effect on ovarian hormones during the middle reproductive years. PMID- 8712196 TI - Does multiple risk factor reduction explain the reduction in fall rate in the Yale FICSIT Trial? Frailty and Injuries Cooperative Studies of Intervention Techniques. AB - In a recent study of fall prevention in 301 community-living older persons (the Yale FICSIT Trial, 1990-1993), participants in the multifactorial targeted intervention (TI) group experienced significantly fewer falls than participants in the social visit (SV) control group. In the present paper, the authors explore the relation between changes in the studied risk factors and the occurrence of falling. In comparison with SV participants, TI participants showed significantly greater improvements in postural blood pressure change (p = 0.01), step length (p = 0.004), use of > or = 4 medications (p = 0.003), and unsafe tub and toilet transfers (p = 0.05), while change in balance was of borderline significance (p = 0.08). Reduction in the occurrence of falling, in turn, was at least marginally associated with improvements in balance, postural blood pressure change, step length, lower extremity strength/range of motion, and transfers. When participants were divided into tertiles based on a composite risk factor change score, a significantly higher percentage of TI participants (42%) than SV participants (22%) were in the greatest risk factor reduction tertile. Among TI participants, there was a progressively lower fall rate per person per year in the tertiles with the least, intermediate, and greatest risk reduction (0.832, 0.624, and 0.260), respectively. A similar but weaker relation between risk factor reduction and fall rate was seen in the SV group. When compared within tertiles, essentially adjusting for the amount of risk factor reduction, the fall rates among TI and SV participants in the greatest risk factor reduction tertile were identical (0.260 falls per person per year), and the rates in the least reduction tertile were similar (0.832 vs. 1.040 falls per person per year); this suggests that risk factor reduction at least partially mediated the treatment effect. These results support the feasibility of implementing and analyzing the effectiveness of a multiple risk factor reduction strategy in the aged. PMID- 8712197 TI - Evidence against the operation of selective mortality in explaining the association between cigarette smoking and reduced occurrence of idiopathic Parkinson disease. AB - To investigate the association between idiopathic Parkinson disease (IPD) and reduced frequency of prior cigarette smoking, the authors compared the 29-year follow-up mortality rates and IPD incidence rates of men who were either cigarette smokers or nonsmokers at the time of enrollment in the Honolulu Heart Study (1965-1968). Based on IPD cases detected up to June 30, 1994, the age adjusted incidence rate in smokers was less than half that in nonsmokers: 34.4 versus 94.2 cases per 100,000 person-years of pre-illness follow-up, respectively. When data were stratified by 5-year age group, lower IPD incidence in smokers was observed at all ages between 50 and 90 years. Age-specific mortality trends for smokers and nonsmokers with and without IPD suggested that increased mortality in IPD patients was mostly associated with IPD itself and not with smoking. The slight excess mortality in smokers without IPD, versus nonsmokers without IPD, appeared insufficient to account for the "missing" incident IPD cases in smokers. These IPD incidence and mortality data are not highly consistent with the "selective mortality" hypothesis, which attributes reduced prior smoking frequency, typically reported by persons with IPD, to accelerated mortality in undiagnosed IPD-affected persons who smoke. The "protective" association of cigarette smoking with IPD occurrence may thus be real, suggesting the need for further study of biologic mechanisms of protection. PMID- 8712198 TI - Risk factors for Toxoplasma gondii infection in pregnancy. Results of a prospective case-control study in Norway. AB - From 1992 to 1994, a prospective case-control study designed to identify preventable risk factors for Toxoplasma gondii infection in pregnancy was conducted in Norway. Case-patients were identified through a serologic screening program encompassing 37,000 pregnant women and through sporadic antenatal testing for Toxoplasma infection. A total of 63 pregnant women with serologic evidence of recent primary T. gondii infection and 128 seronegative control women matched by age, stage of pregnancy, expected date of delivery, and geographic area were enrolled. The following factors were found to be independently associated with an increased risk of maternal infection in conditional logistic regression analysis (in order of decreasing attributable fractions): 1) eating raw or undercooked minced meat products (odds ratio (OR) = 4.1, p = 0.007); 2) eating unwashed raw vegetables or fruits (OR = 2.4, p = 0.03); 3) eating raw or undercooked mutton (OR = 11.4, p = 0.005); 4) eating raw or undercooked pork (OR = 3.4, p = 0.03); 5) cleaning the cat litter box (OR = 5.5, p = 0.02); and 6) washing the kitchen knives infrequently after preparation of raw meat, prior to handling another food item (OR = 7.3, p = 0.04). In univariate analysis, travelling to countries outside of Scandinavia was identified as a significant risk factor, but this variable was not independently associated with infection after data were controlled for factors more directly related to the modes of infection. PMID- 8712199 TI - Estimating the prevalence of drug use from self-reports in a cohort for which biologic data are available for a subsample. AB - Diagnostic procedures, used singly or in combination, are crucial in the determination of the presence and prevalence of medical and other conditions. In the absence of a "gold standard," two or more measures or diagnostic tests are often available that may be used to estimate true prevalence. The authors have developed a statistical method with which to calculate more precise estimates of a condition in the presence of two diagnostic measures, one measurement being performed on the entire study sample and a second, more precise one being made in a random sample of the study sample. This method uses the well-known equations which express the probabilities of the four possible outcomes of the two measures in terms of the sensitivities and specificities of the measures and the prevalence of the condition and some properties of maximum likelihood estimates to obtain an expression for the estimated true prevalence and its precision. The method is illustrated by applying it to data collected by urinalysis and self report in 1992-1993 in a national multisite study-the Cocaine Treatment Outcome Study. Through application of this methodology, a more precise estimate of the true prevalence of substance use can be obtained from two measures, one biologic and the other self-reported. Detailed equations and expressions are provided so that the method can be applied in other situations where diagnostic data from two different sources or procedures are available. PMID- 8712200 TI - Collapsing ordered outcome categories: a note of concern. AB - When analyzing and interpreting data from an epidemiologic study where ordinal (ordered categorical) outcomes have been measured in different exposure groups, an effect parameter of interest is the common odds ratio implied by the proportional odds model. This model can sometimes be applied to a collapsed outcome variable, instead of the measured variable, without reducing efficiency considerably. However, in a given data set, changing the outcome categories can affect the effect estimate as well as the inference being drawn from the data, even if the true effect itself has not changed. In particular, one should be careful in dichotomizing the measured outcome variable. PMID- 8712201 TI - Intraclass correlation estimates in a school-based smoking prevention study. Outcome and mediating variables, by sex and ethnicity. AB - Most school-based smoking prevention studies employ designs in which schools or classrooms are assigned to different treatment conditions while observations are made on individual students. This design requires that the treatment effect be assessed against the between-school variance. However, the between-school variance is usually larger than the variance that would be obtained if students were individually randomized to different conditions. Consequently, the power of the test for a treatment effect is reduced, and it becomes difficult to detect important treatment effects. To assess the potential loss of power or to calculate appropriate sample sizes, investigators need good estimates of the intraclass correlations for the variables of interest. The authors calculated intraclass correlations for some common outcome variables in a school-based smoking prevention study, using a three-level model-i.e., students nested within classrooms and classrooms nested within schools. The authors present the intraclass correlation estimates for the entire data set, as well as separately by sex and ethnicity. They also illustrate the use of these estimates in the planning of future studies. PMID- 8712202 TI - Re: "Sexually transmitted diseases, sexual behavior, and cocaine use in inner city women". PMID- 8712203 TI - Brain swelling after dialysis: old urea or new osmoles? AB - The pathogenesis of brain swelling and neurological deterioration after rapid hemodialysis (dialysis disequilibrium syndrome) is controversial. The "reverse urea hypothesis" suggests that hemodialysis removes urea more slowly from the brain than from the plasma, creating an osmotic gradient that results in cerebral edema. The "idiogenic osmole hypothesis" proposes that an osmotic gradient between brain and plasma develops during rapid dialysis because of newly formed brain osmoles. In this review, the experimental basis for the two hypotheses are critically examined. Based on what is known about the physiology of urea and water diffusion across the blood-brain barrier, and empiric observations of brain solute composition after experimental hemodialysis, we conclude that the "reverse urea hypothesis" remains a viable explanation for dialysis disequilibrium and that rapid reduction of a high urea level in and of itself predisposes to this condition. PMID- 8712204 TI - Long-term bone loss in kidney transplant recipients: a cross-sectional and longitudinal study. AB - Organ transplantation is associated with an early bone loss that subsequently increases the risk of osteopenia and bone fractures. It is not known whether bone loss continues in long-term survivors, but persistent bone demineralization could further jeopardize an already diminished bone mass. To better define long-term bone status of kidney transplant recipients (KTR), we conducted cross-sectional and longitudinal evaluations of bone mineral density (BMD) in 70 KTR with a mean posttransplantation time of 8.1 years. BMD was determined by dual-energy X-ray absorptiometry and was repeated in 55 of the patients after a mean follow-up period of 22 +/- 5 months. Lumbar and femoral osteopenia, defined as a BMD lower than 2 standard deviations from mean value of sex- and age-matched controls, was present in 28.6% and 10.5% of patients, respectively. There was a significant negative correlation between cumulative prednisone dose and adjusted lumbar as well as femoral BMD (R = 0.45, P < 0.001 and R = 0.29, P < 0.05, respectively). Five patients had a vertebral BMD below a fracture threshold of 0.777 g/cm2. Vertebral fractures (VF) were found in four men and were associated with higher prednisone dosage (P < 0.05), larger cumulative prednisone dose (P < 0.05), and significantly lower BMD values. According to World Health Organization recent criteria for women, prevalences of lumbar and femoral osteopenia and lumbar and femoral osteoporosis in female patients reach 75%, 65%, 33%, and 10%, respectively. The longitudinal part of the study showed a persistent pathological lumbar demineralization process. Over the study period, BMD, expressed as a percentage of that of controls, decreased from 89 +/- 14% to 86 +/- 14% (P < 0.001). Annual change of bone mass was -1.7 +/- 2.8% per year. Accelerated vertebral bone loss (defined as a decrease of BMD, expressed as a percentage of that of controls, of more than 1% per year) was present in 56% of patients and was associated with higher prednisone dosage (P < 0.01). In conclusion, although VF are relatively infrequent in long-term survivors of renal transplantation, osteopenia is a frequent finding, and a substantial proportion of women present lumbar osteoporosis. An ongoing demineralization process of the spine is also demonstrated and probably contributes to long-term spinal osteoporosis incidence. Prednisone dosage remains the most constantly isolated risk factor. PMID- 8712205 TI - Effect of angiotensin II on the renal response to amino acid in rats. AB - Administration of the nitric oxide (NO) synthase blocker, N(G)-monomethyl L arginine prevents the increase in glomerular filtration rate (GFR) normally observed with glycine, an effect that is restored by angiotensin II (AII) blockers. These findings suggest that changes in NO and AII dictate the presence or absence of renal vasodilation during amino acid (AA) infusion. We examined the effect of branched-chain (BCAA) and non-branched-chain (NBCAA) AA on GFR, NO, and AII to determine if abnormal NO or AII responses could explain the absence of vasodilation with BCAA. Our findings demonstrated that NBCAA increased GFR and NO and did not modify AII, either plasma (AIIp) or kidney (AIIk) AII. The response with BCAA was strikingly different. L-Valine increased GFR without modifying NO or AII. L-Leucine increased AIIk and NO but did not increase GFR. Administration of AII blockers (captopril or losartan) was associated with an increase in GFR during infusion of leucine. Single nephron studies demonstrated that increased AIIk with L-leucine was associated with decreased absolute proximal reabsorption and probably activation of the tubuloglomerular feedback. An AA-specific increase in AIIk is critical to inhibition of the normal renal response to AA infusion. NO generation is an important mediator but not the sole mechanism that determines the increase in GFR during amino acid infusion. PMID- 8712206 TI - Acute tubulointerstitial nephritis and uveitis with antineutrophil cytoplasmic antibody. AB - We report a case of acute tubulointerstitial nephritis and uveitis (TINU syndrome) in a 47-year-old woman who also was found to have antineutrophil cytoplasmic antibody. The patient developed severe acute renal failure that improved after a high dose (1 mg/kg/d) of corticosteroid therapy. The diagnosis of the disorder is discussed, as well as the finding of antineutrophil cytoplasmic antibody. PMID- 8712207 TI - Monoclonal heavy chain (immunoglobulin G3) deposition disease: report of a case. AB - Seven cases of nonamyloid heavy chain (gamma chain) deposition disease have been previously reported. We describe one case of a 79-year-old woman presenting with proteinuria and microscopic hematuria whose renal biopsy showed nodular glomerulosclerosis with deposition of gamma3 heavy chains and complement in the glomeruli and tubular basement membranes with no associated light chain deposition. The patient did not have multiple myeloma. This case is unique in that in all previously reported cases of heavy chain deposition disease the gamma chain subtype has been either gamma1 or gamma4. PMID- 8712208 TI - Midodrine efficacy and pharmacokinetics in a patient with recurrent intradialytic hypotension. AB - Recurrent intradialytic hypotension, a complication of hemodialysis, is a consequence of an inadequate compensatory response or a paradoxic response to ultrafiltration-induced volume reduction. We report the use of midodrine, an alpha agonist, in an 18-year-old man with Bardet-Biedl syndrome and recurrent intradialytic hypotension. The clinical features of the intradialytic hypotensive spells are consistent with a paradoxic withdrawal of sympathetic activity, although an underlying abnormality in autonomic dysfunction cannot be excluded. Midodrine significantly increased the intradialytic blood pressure and decreased the intradialytic hypotensive episodes requiring intervention. The pharmacokinetic characteristics of the prodrug midodrine and the active metabolite de-glymidodrine in this patient with end-stage renal disease approximate those reported for patients with normal renal function. However, the prolonged terminal half-life for the active metabolite, de-glymidodrine, warrants careful administration in patients with renal failure. PMID- 8712209 TI - Subcutaneous phaeohyphomycosis in a renal transplant recipient: a case report and review of the literature. AB - Subcutaneous phaeohyphomycosis caused by dematiaceous fungi has rarely been described in immunocompromised patients. We report a case of subcutaneous infection caused by Phialophora parasitica in a renal transplant recipient. The role of early identification of the offending fungal species followed by a wide local excision in the management of such patients has been discussed. PMID- 8712210 TI - Superoxide dismutase activity in human glomerulonephritis. AB - Superoxide dismutase (SOD) in renal tissue biopsy specimens obtained from patients with immunoglobulin A nephropathy (13 cases) and non-immunoglobulin A mesangial proliferative glomerulonephritis (nine cases) was studied at the protein level by an enzyme-linked immunosorbent assay method and at the mRNA level by the reverse transcriptase-polymerase chain reaction (RT-PCR) assay. Total SOD activity in the tissue supernatant was measured by applying an electron paramagnetic resonance/spin trapping method. Normal renal tissues obtained from kidneys removed for malignancies (six cases) were included as healthy controls. The copper and zinc form of SOD (Cu,Zn-SOD) activity at both the protein and mRNA levels was lower in the moderately or severely damaged tissues compared with that in the normal or mildly damaged tissues. On the other hand, manganese SOD (Mn SOD) values at either the protein level or the mRNA level did not differ significantly between control and patient samples. In the histochemical study using a polyclonal rabbit anti-Cu,Zn-SOD antibody, the staining intensity for Cu,Zn-SOD antigen was lower in the areas with advanced histologic damage than in the intact tissues. A follow-up study showed that renal function deterioration was proportionately slower in patients whose SOD activity was within the range of healthy tissue levels at the time of biopsy. Our data suggest that a lower level of SOD activity, whether as a cause or a consequence of the disease process, might induce a decrease in the scavenger reaction of superoxide (O2-) thus causing the tissue to become more vulnerable to oxidative stress. PMID- 8712211 TI - Mitochondrial genome mutations and kidney disease. AB - Mutations in the mitochondrial genome have been shown to be responsible for several neuromuscular diseases in humans. In this article, we discuss the molecular genetics of mitochondria, their centrality in cellular energy production, and reasons why their genome is extremely vulnerable to mutation. Mitochondrial DNA (mtDNA) mutations and their classic encephalomyopathic clinical phenotypes are briefly reviewed, and evidence presented that mtDNA mutations also present primarily as kidney diseases. Research trends in the field are discussed. Suggestions are made regarding future work, the clinical implications thereof, and potential therapeutic utility accruing from these advances. PMID- 8712212 TI - Foregoing renal dialysis: a case study and review of ethical issues. The End Stage Renal Disease Group. AB - End-stage renal disease is treated by long-term dialysis when renal transplantation is not feasible. At a late stage in the disease, for a variety of reasons, dialysis is frequently stopped. This is one of the most common causes of death in dialysis patients. Ethical issues related to withdrawal of dialysis are therefore commonly encountered in nephrology practices. A case study involving discussions to forego dialysis in an incompetent patient is presented, and the ethical issues raised by this case, particularly related to the concepts of medical futility, informed choice, and justice are discussed. Finally, procedural approaches are suggested that will help to address the ethical problems raised and to assist in decision making at the time that the discontinuation of dialysis is being considered. PMID- 8712213 TI - Looking upon the water. Isn't truth the safest lie? PMID- 8712214 TI - L-carnitine and erythropoietin requirements in hemodialysis patients. PMID- 8712215 TI - "PD Plus": is it a new concept? PMID- 8712216 TI - Falsely elevated serum vancomycin concentrations. PMID- 8712217 TI - Possible discrimination of Gitelman's syndrome from Bartter's syndrome. PMID- 8712218 TI - Cyclosporine enhances leukocyte adhesion to vascular endothelium under physiologic flow conditions. AB - We investigated the effect of cyclosporine (CyA) on leukocyte adhesion to endothelium under flow conditions. Confluent human umbilical vein endothelial cells (HUVECs) were incubated for 24 hours with CyA (1, 5, and 10 micromol/L) and then exposed to a total human leukocyte suspension in a parallel plate flow chamber under laminar flow (1.5 dynes/cm2. Human umbilical vein endothelial cells stimulated with interleukin-1beta (20 U/mL) were used as a positive control. Adherent cells were measured by digital image analysis. Results showed that CyA dose-dependently increased the number of leukocytes adhering to HUVECs compared with control cells. Leukocyte adhesion markedly increased on HUVECs incubated with interleukin-lbeta, one of the most potent inducers of endothelial cell adhesiveness. Exposure of endothelial cells to CyA did not affect the number of rolling leukocytes, which was similar to control values. To examine the role of adhesion molecules in CyA-induced leukocyte adhesion, HUVECs were incubated with monoclonal antibodies against intercellular adhesion molecule-1 (ICAM-1), vascular adhesion molecule-1 (VCAM-1), and E-selectin before adhesion assay. Functional blocking of ICAM-1, VCAM-1, and E-selectin on endothelial cells significantly inhibited CyA (10 micromol/L)-induced leukocyte adhesion. Confocal fluorescence microscopy studies showed that CyA induced an increase in the endothelial surface expression of ICAM-1, VCAM-1, and E-selectin. Pretreatment of leukocytes with the platelet activating factor receptor antagonist L659,989 significantly reduced the number of leukocytes adhering to CyA-treated HUVECs. We suggest that CyA enhances leukocyte adhesion to endothelium by upregulating adhesive proteins on endothelial surface membrane. Blocking leukocyte receptor for platelet-activating factor partially prevents adhesion, suggesting a role for endothelial cell-associated platelet-activating factor in the interaction between leukocytes and CyA-treated endothelium. PMID- 8712219 TI - Increased mRNA expression of metalloproteinase-9 in peripheral blood monocytes from patients with immunoglobulin A nephropathy. AB - We examined metalloproteinase (MMP)-1, -2, -3, and -9 mRNA expression by peripheral blood monocytes from 50 patients with immunoglobulin A (IgA) nephropathy, 20 with membranous nephropathy, 10 with minimal-change nephrotic syndrome, five with focal glomerulosclerosis, 30 with non-IgA proliferative glomerulonephritis, and 40 healthy normal controls who were comparable with regard to age and sex. Monocytes from patients with IgA nephropathy expressed a higher level of MMP-9 mRNA than those from patients with other forms of glomerulonephritis or from healthy controls (MMP-9 to glyceraldehyde-3-phosphate dehydrogenase ratio: IgA nephropathy, 1.68 +/- 0.24; membranous nephropathy, 0.22 +/- 0.08; minimal-change nephrotic syndrome, 0.24 +/- 0.06; focal glomerulosclerosis, 0.32 +/- 0.08; non-IgA proliferative glomerulonephritis, 0.30 +/- 0.12; and healthy controls, 0.16 +/- 0.04). When the biopsy specimens were classified into four grades according to the severity of glomerular and interstitial pathology, highly significant differences were observed among MMP-9 mRNA levels in monocytes from all four groups of patients with IgA nephropathy (grade I, 0.44 +/- 0.09; grade II, 1.06 +/- 0.26; grade III, 2.22 +/- 0.68; grade IV, 2.86 +/- 0.88). In addition, MMP-9 mRNA levels from patients with IgA nephropathy correlated with urinary protein excretion (P < 0.001). However, we detected minimal mRNA expression of MMP-1, -2, and -3 by peripheral blood monocytes from patients with IgA nephropathy or other forms of glomerulonephritis and from normal healthy controls. Our results suggest that increased MMP-9 mRNA expression in circulating monocytes may contribute to the progression of IgA nephropathy. PMID- 8712220 TI - Intercellular adhesion molecule-1/leukocyte function associated antigen-1 mediated and complement receptor type 4-mediated infiltration and activation of glomerular immune cells in immunoglobulin A nephropathy. AB - Glomerular expression of intercellular adhesion molecule-1 (ICAM1) (CD54) and membrane cofactor protein (MCP; CD46) and positive infiltrating cells in leukocyte function associated antigen-1 (LFA1)alpha (CD11a) and C3bi receptors (CR3/CD11b, CR4/CD11c) were examined by the indirect immunoperoxidase method on 43 sets of repeated renal biopsy specimens from patients with immunoglobulin A nephropathy. Twenty-four-hour urine protein at the time of renal biopsy was also evaluated. Glomerular infiltration of LFA1alpha+ cells was significantly correlated with glomerular expression of ICAM1 (r = 0.494, P < 0.0001). Glomerular complement receptor type 4 (CR4)+ cells were significantly correlated with glomerular expression of MCP (r = 0.405, P < 0.0001). The glomerular expressions of ICAM1 and MCP were significantly correlated with each other (r = 0.700, P < 0.00001). The glomerular infiltrations of LFA1alpha+ and CR4+ cells were highly correlated with each other (r = 0.884, P < 0.00001), and both cell types were significantly correlated with urine protein (respectively, r = 0.426 and 0.478, P < 0.001 and 0.0001). When the change in these parameters between the time of the initial and follow-up biopsies was evaluated, there was a significant correlation between the change in glomerular expression of ICAM1 (DeltaICAM1) and MCP (DeltaMCP) as well as between the change in glomerular infiltration of LFA1alpha+ cells (DeltaLFA1alpha+) and CR4+ cells (DeltaCR4+). Both DeltaLFA1alpha+ and DeltaCR4+ were significantly correlated with the change in urine protein. These findings suggest that ICAM1/LFA1 interaction and MCP mediated C3bi/C3biR interaction cooperate and participate in persistent glomerular infiltration of immune cells in immunoglobulin A nephropathy, and that these LFA1alpha+ and C3biR+ cells contribute to the induction of proteinuria. PMID- 8712221 TI - Timing of complications in percutaneous renal biopsy: what is the optimal period of observation? AB - Percutaneous renal biopsies of native kidneys were performed in 394 adult patients (aged > or = 15 years) at Rush-Presbyterian-St Lukes Medical Center between February 1983 and April 1995. All biopsies were performed with the use of real-time ultrasound; the last 169 were done with the use of an automated biopsy needle. All patients had a normal bleeding time and were observed for 23 to 24 hours postbiopsy. A biopsy-related complication occurred in 52 patients (13%). Minor complications, defined as gross hematuria, or perinephric hematoma that resolved without the need for transfusion or intervention, occurred in 26 patients (6.6%). Major complications, defined as those requiring a transfusion, invasive procedure, or resulting in septicemia, also occurred in 26 patients (6.6%). Patients with complications did not differ from those without complications at the time of biopsy with respect to age, blood pressure, serum creatinine, or bleeding time, but did have a lower prebiopsy hemoglobin level (11 +/- 2 g/dL v 12 +/- 2 g/dL; P < 0.05). No significant difference in age, blood pressure, serum creatinine, or hemoglobin level at the time of biopsy was observed in patients with major complications compared with those with minor complications. The time interval between the procedure and the identification of a complication was available for 44 patients (85%): 24 (92%) had a major complication and 20 (77%) had a minor complication. The complication was apparent within 24 hours in all but one patient (98%). Overall the complication was identified in only 52% of patients at < or = 4 hours, 77% at < or = 8 hours, and 95% at < or = 12 hours. Major complications were identified in 46% of patients at < or = 4 hours, 79% at < or = 8 hours, and 100% at < or = 12 hours. In patients with major complications the postbiopsy hemoglobin decreased by 3.2 +/- 1.8 g/d; this was significantly greater (P < 0.0001) than the decrease seen in patients with minor complications (1.2 +/- 1.0 g/dL) or in patients with no complications (0.9 +/- 0.8 g/dL). When considering a percutaneous renal biopsy as an outpatient procedure, observation of patients for 23 to 24 hours is optimal. Observation for 8 hours or less risks missing > or = 20% of complications. PMID- 8712222 TI - The impact of anemia on cardiomyopathy, morbidity, and and mortality in end-stage renal disease. AB - To determine the possible association between anemia and clinical and echocardiographic cardiac disease, a cohort of 432 end-stage renal disease patients (261 on hemodialysis and 171 on peritoneal dialysis) who started dialysis therapy between 1982 and 1991 were followed prospectively for an average of 41 months. Baseline demographic, clinical, and echocardiographic assessments were performed, as well as monthly serial clinical and laboratory tests while the patients were on dialysis therapy. The mean (+/-SD) hemoglobin level during dialysis therapy was 8.8 +/- 1.5 g/dL. After adjusting for age, diabetes, and ischemic heart disease, as well as for blood pressure and serum albumin levels measured serially, each 1 g/dL decrease in mean hemoglobin was independently associated with the presence of left ventricular dilatation on repeat echocardiogram (odds ratio, 1.46; P = 0.018) and the development of de novo (relative risk [RR] = 1.28; P = 0.018) and recurrent (RR = 1.20; P = 0.046) cardiac failure. In addition, each 1 g/dL decrease in the mean hemoglobin level was independently associated with mortality while the patients were on dialysis therapy (RR = 1.14; P = 0.024). Anemia had no independent association with the development of ischemic heart disease while the patients were on dialysis therapy. Anemia, an easily reversible feature of end-stage renal disease, is an independent risk factor for clinical and echocardiographic cardiac disease, as well as mortality in end-stage renal disease patients. PMID- 8712223 TI - Accumulation of glycated albumin in end-stage renal failure: evidence for the principle of "physiological microalbuminuria". AB - In light of the growing understanding of the toxic effects of glycated albumin and of the preferential excretion of this substance, the excretion of glycated albumin could be considered a physiologic function of the kidney. Furthermore, if the increased load of glycated albumin in diabetic patients results in glycated albumin excretion rates in the range of 20 to 200 microg/min, might this not be considered "physiologic microalbuminuria"? The hypothesis is presented that microalbuminuria composed of glycated albumin is a homeostatic renal function. Although some proteins are glycosylated for their normal physiologic function, many proteins are glycated nonenzymatically according to ambient blood glucose. Albumin is subject to nonenzymatic glycation in all humans, but at increased rates in diabetic patients. Glycated albumin induces changes in the microvasculature and glomerulus that may lead to endothelial dysfunction and diabetic nephropathy, respectively. Renal excretion of glycated albumin is enhanced compared with native albumin. To explore this potential homeostatic function of the kidney, patients with impaired renal function were studied to determine whether glycated albumin accumulates. Plasma levels of glycated albumin were determined in diabetic and nondiabetic patients on hemodialysis. Hemoglobin A1c was used as an index of the rate of nonenzymatic glycation of proteins. Hemoglobin A1c was increased in the diabetic subjects but was normal in the nondiabetic group (7.9% +/- 0.5% v 6.2% +/- 0.2%, respectively; P < 0.01). On the other hand, the glycated albumin was elevated in both groups and was not significantly different between them (1.95% +/- 0.15% in the diabetic patients v 1.75% +/- 0.14% in the nondiabetic patients; P = NS). The results of this study provide the first clinical evidence supporting the hypothesis that the excretion of glycated albumin is a homeostatic renal function. PMID- 8712224 TI - Most nephrologists are primary care providers for chronic dialysis patients: results of a national survey. AB - The medical care of end-stage renal disease (ESRD) patients includes not only dialysis-related medical care but preventive and general medical care as well as the care of minor acute illnesses. There is little information about nephrologists' interpretation of their potential role as a primary health care provider for the general medical needs of chronic dialysis patients. To characterize nephrologists' primary care practice patterns related to the care of chronic dialysis patients, we surveyed a randomly selected group of practicing nephrologists and asked questions about preventive medicine guidelines followed, treatment of minor acute illnesses, and management of chronic medical problems in ESRD patients. The results of 233 questionnaires (46% response rate) were analyzed. Most of the responding nephrologists were men (91%), were board certified in internal medicine (96%) and nephrology (83%), were out of nephrology practice for a mean of 16 years, had a mean age of 48 +/- 7 years, and were in private practice (65%). The average percentage of time spent with chronic dialysis patients was reported as 30%; 38% of that time was devoted to the general medical care of those patients. Ninety percent of nephrologists reported that they provided primary care to their dialysis patients, and only 21% said a nurse practitioner or physician assistant worked with them. Age and number of years in practice were the only demographic factors increasing the likelihood of nephrologist-provided primary care, with older, more experienced practitioners more likely to be providing primary care to dialysis patients. Most nephrologists reported that they managed minor acute illnesses and comorbid conditions (diabetes mellitus, cardiac disease, and gastrointestinal disease) in their dialysis patients. Nephrology fellowship training programs and recertification programs may need to address issues of primary general health care of ESRD patients. Plans under development for health care programs and reimbursement criteria also need to recognize and consider the primary medical care role practiced by nephrologists caring for ESRD patients. PMID- 8712225 TI - Survival of patients undergoing renal replacement therapy in one center with special emphasis on racial differences. AB - This study compared racial differences in end-stage renal disease (ESRD) in 550 patients starting renal replacement therapy at a large academic dialysis center between January 1, 1990, and December 31, 1993, with follow-up through December 31, 1994. Patient groups were compared with respect to cause of ESRD, comorbid factors at the start of dialysis therapy, choice of modality, transplantation rate, and survival. Fifty-eight percent of the patients were white and 42% were African-American. There was a similar distribution of causes of ESRD between races. African-American patients were less likely to choose peritoneal dialysis as initial therapy (11.6% v 29.3%; P < 0.001) and were less likely to change dialysis modality. Transplantation rates were significantly different between African-American and white patients (9.3% v 27.6%; P < 0.001). African-Americans less frequently received living-related, living-nonrelated, and cadaveric renal transplants. Given differences in transplantation rates and in survival of transplanted patients versus patients on dialysis, survival analysis was performed without censoring for transplantation. A multivariate Cox proportional hazards model was formed, and the following were identified as being significant independent predictors of survival: age, race, age-race interaction, serum albumin at the start of dialysis, activity level at the start of dialysis, and presence of congestive heart failure and cancer. Age had little effect on survival among African-American patients, while it was a significant predictor of survival in white patients. In the group of patients starting dialysis before the age of 30 years, African-American patients had a significantly increased mortality risk compared with white patients. However, white patients older than 50 years had a higher mortality risk; this risk difference increased with age. Racial differences in mortality among older white patients could not be explained by differences in comorbid conditions, transplantation rates, or withdrawal from dialysis. PMID- 8712226 TI - Heparin-induced thrombocytopenia in hemodialysis patients. AB - This study was conducted to determine the incidence of heparin-induced thrombocytopenia (HIT), an important complication in heparin therapy, in 154 hemodialysis patients, with characterization of the subtypes using an enzyme linked immunosorbent assay. The etiology of the immunologic type of HIT is suggested to involve the binding of a specific antibody for platelet factor 4 heparin complex to platelets and their consequent activation. The 154 consecutive patients were newly treated with hemodialysis due to chronic or acute renal failure between January 1993 and July 1995. Heparin-induced thrombocytopenia was suspected in six patients (3.9%), and its presence was confirmed by platelet aggregation testing. Five of the patients with HIT had anti-platelet factor 4 heparin complex antibody detected by the enzyme-linked immunosorbent assay and were diagnosed with immunologic HIT. The patient who did not have the antibody was thought to have another type of HIT. PMID- 8712227 TI - Fungal peritonitis in a large chronic peritoneal dialysis population: a report of 55 episodes. AB - Fungal peritonitis (FP) is a rare but serious complication of chronic peritoneal dialysis (CPD) therapy and is associated with high morbidity and CPD drop-out. Risk factors and management of FP remain controversial. We reviewed our experience with FP in an attempt to identify risk factors and to examine outcome in relation to treatment strategies. Between March 1984 and August 1994, 704 patients were maintained on CPD therapy in our unit. A total of 1,712 episodes of peritonitis were identified among these patients. Fungal peritonitis accounted for 55 (3.2%) of these episodes. The patients on CPD therapy who developed FP were similar to those who did not develop FP with regard to age, gender, underlying etiology for end-stage renal disease, and comorbid disease. Prior antibiotic use was noted in 87.3% of episodes of FP. The peritonitis rate in the patients who developed FP was one episode every 5.1 months compared with one episode every 9.9 patient-months in the CPD patients who did not develop this infection. Candida sp caused 74.5% of the episodes of FP. All patients were treated with antifungal drugs. In 85.5% of infections the Tenckhoff catheter was removed within 1 week of the diagnosis of FP; 31.9% of the patients who had the Tenckhoff catheter removed did not have the catheter replaced because of death or transfer to hemodialysis. In the patients who developed FP, 68.1% had the Tenckhoff catheter replaced; of these patients, 90.6% and 59.4% were on CPD therapy 1 and 6 months after catheter replacement, respectively. We conclude that risk factors identified in our population include peritonitis rate and prior antibiotic use. Fungal peritonitis is rare in our CPD population, and although it leads to significant CPD drop-out, it can be managed in many patients with antifungal therapy, early catheter removal, and temporary hemodialysis. Of the catheters replaced between 2 and 8 weeks after the diagnosis of FP, 91% functioned successfully, allowing continuation of CPD. PMID- 8712228 TI - Pretransplant rejection risk assessment through enzyme-linked immunosorbent assay analysis of anti-HLA class I antibodies. AB - Soluble HLA enzyme-linked immunosorbent assays (ELISAs) for the detection of anti HLA class I immunoglobulin G (IgG), IgG1, IgG2, IgG3, IgG4, IgM, and IgA antibodies were developed and used to analyze retrospectively the correlation between pretransplant allosensitization and posttransplant rejection episodes in renal allograft recipients. Enzyme-linked immunosorbent assay plates were coated with 46 different soluble HLA preparations representing 40 different HLA class I antigens. After incubation with a serum specimen, bound antibodies were detected with a peroxidase-conjugated antibody. Serum specimens from 85 patients were analyzed. All patients tested positive by microlymphocytotoxicity (ie, >5% panel reactive antibody [PRA]). Approximately half (56%) of the patients had experienced one or more rejection episodes within 12 months posttransplantation. Fifty-five patients tested positive by ELISA (total IgG %PRA >10%). A strong correlation between first-year rejection and ELISA-detected anti-HLA class I IgG1 was observed (P = 0.0004). The predictive value for IgG1 and first-year rejection was 77.5%, demonstrating that ELISA results identify patients at high risk of rejecting the transplanted kidney. Anti-HLA class I total IgG detected by ELISA also correlated with first-year rejection episodes (P = 0.04). The presence of anti-HLA class I IgG2, IgG3, IgG4, or IgM was not predictive of first-year rejection episodes. Anticlass I IgA antibodies were only found in combination with anti-class I IgG1 antibodies. PMID- 8712229 TI - Recurrence of immunoglobulin A nephropathy after renal transplantation in the cyclosporine era. AB - Immunoglobulin A nephropathy (IgAN) frequently recurs in patients after renal transplantation (RT) on a conventional regimen of immunosuppressive therapy, but little is known about the influence of cyclosporine (Cs) on such a recurrence. We studied 84 patients retrospectively who underwent RT for renal failure attributable to IgAN (n = 71) or Henoch-Schonlein purpura nephropathy (HSPN) (n = 13) in two transplantation units, between January 1985 and June 1991 and were treated with Cs. Four patients died 3 months to 8 years after RT. Graft survival was 88% at 1 year, 75.2% at 5 years, and 63% at 8 years. Fifty patients underwent at least one graft biopsy, but studies with immunofluorescence were performed on only 28 (23 IgAN and 5 HSPN). After a mean follow-up of 68.1 +/- 37.2 months, mesangial IgA deposits recurred in 13 of the 28 patients (12 IgAN and 1 HSP) (prevalence, 46.4%). Among the 13 patients with recurrence of IgA deposits, all but 4 had urinary abnormalities. Light microscopy showed mesangial deposits and focal and segmental glomerular changes in 9 cases. Four patients lost their graft function 69 to 119 months after RT, and 2 had severe graft dysfunction. The rates of graft failure and mean serum creatinine at 1, 5, and 8 years were similar in the 13 patients with recurrence and the 15 patients without proven recurrence. In conclusion, Cs did not reduce the incidence or severity of IgAN recurrence. The latter was the cause of graft loss or dysfunction in 46.1 % of the patients with recurrent IgA deposits. Recurrent glomerulonephritis did not influence the 8-year graft survival in patients with IgAN or HSPN, but it may be an important cause of graft loss as evidenced by more extended follow-up. PMID- 8712230 TI - Revisiting digoxin toxicity. PMID- 8712231 TI - Be on the lookout for skin cancer. PMID- 8712232 TI - Are you maximizing your potential? PMID- 8712233 TI - Minimizing meperidine use. PMID- 8712234 TI - Promote, don't protect. PMID- 8712235 TI - Protecting your patients from VRE. PMID- 8712237 TI - Diabetes insipidus. PMID- 8712236 TI - Advance directives. Most patients don't have one. Do yours? PMID- 8712238 TI - How to improve your shift report. PMID- 8712239 TI - Controlling diarrhea in the HIV patient. PMID- 8712240 TI - Back to basics assessing orthostatic vital signs. PMID- 8712241 TI - Emergency! Gasoline exposure. PMID- 8712243 TI - More than rearranging the deck chairs. PMID- 8712242 TI - How to transfer patients to other units. PMID- 8712244 TI - Nationally speaking. Life history and narrative research: generating a humanistic knowledge base for occupational therapy. AB - As a profession, occupational therapists are guided in practice by the accumulated knowledge of occupational therapy. This article demonstrates the contributions of life-history and narrative research to this knowledge base. We are suggesting that in response to our humanistic roots, we must pursue additional knowledge, principles of practice, and ethical philosophies that support practice. We have argued that our ideologic concern for the client must guide our choice of epistemologies to investigate the lived experience to those whom we serve. "What is at stake here is the ethic that is embedded in the epistemology that gives rise to kinds of research" (Brock, 1995, p.157) that meet the societal demands for professionalism and support a humanistic practice. PMID- 8712245 TI - Life histories in occupational therapy clinical practice. AB - This article defines and compares several narrative methods used to describe and interpret patients' lives. The biographical methods presented are case histories, life-charts, life histories, life stories, assisted autobiography, hermeneutic case reconstruction, therapeutic employment, volitional narratives, and occupational storytelling and story making. Emphasis is placed the clinician as a collaborator and interpreter of the patient's life through ongoing interactions and dialogue. PMID- 8712246 TI - Alma: coping with culture, poverty, and disability. AB - This article raises questions about the ways culture affects the nature of health care services. By examining the life story of Alma, a Central American woman who has a daughter with disabilities; her interactions with health care providers; and my own assumptions about cultural differences, I note the impact of cultural differences on coping and adaptation in Alma and in the health care system when working with poor, non-English-speaking clients. PMID- 8712247 TI - The other side of the coin: growing up with a learning disability. AB - This study is a narrative analysis of the life history of Dale, a 25-year-old man with a learning disability who received sensory integrative treatment in early childhood. The analysis revealed that Dale's self likely developed through occupational engagement with three interconnected themes: "my mother said" and "parental tricks"; "adrenaline surges" and "hell-raising"; and work, "deal making," and self-construction. These themes depict Dale's self-construction, character development, occupation selection, and adaptation. In addition, these themes revealed how his parents used principles based on his sensory needs and learning problems to organize his childhood occupations and assist with his adaptation. The concept of occupational metamorphosis is also proposed to describe a person's search for and selection of occupations that satisfy biological and sensory needs. This study supports the idea that the complex nature of adaptation can be revealed through an occupational history, and that adaptation is multifaceted and not absolute. PMID- 8712248 TI - The story of Maricela and Miguel: a narrative analysis of dimensions of adaptation. AB - OBJECTIVES: A mother-child life history was analyzed to examine the multiple dimensions of adaptation in a family grouping, including temporality, maternal values, and life contexts. METHOD: In-depth interviews, participant observation, and documents (e.g., therapy notes, medical records) produced the data for this study. A multiple step narrative analysis included narrative shaping, analysis of major life turnings, and macrostructural analysis of the progress toward life goals. RESULTS: This mother-child life history portrays the challenging life events of a Mexican-origin mother, Maricela, seeking care and assistance to further the development of her son with disabilities, Miguel. Major turnings in her life had both costs and benefits for her in the present and future. Analysis of her multiple roles of mother, professional, daughter, lover-wife, and spiritual devotee revealed that Maricela's life choices, driven by maternal values, diminished the achievement of her personal goals, except those related to her maternal and spiritual roles. Maricela chose actions to realign current and future happenings with her desired life trajectory for her son, which instead of enhancing life conditions for the family often involved short-term and long-term costs. CONCLUSIONS: This mother-child life history demonstrates that a series of moral, relational, and circumstantial factors influence a mother's projected life courses for herself and her child. Adaptation appears to be a dynamic process of realigning life paths to desired life courses, with success evaluated not in a microcosm of time, but from a larger view as these actions contribute to the achievement of desired life goals within a constellation of the person's life goals. PMID- 8712249 TI - Transformative narratives: from victimic to agentic life plots. AB - Basic to occupational engagement is a person's personal power to author choices. Impairment in functioning moves some clients from an agentic identity of self to a victimic identity. The change in identity causes previously self-directed clients to adopt a passive and acquiescent stance toward their lives. The recovery of occupational functioning includes the restoration of the person's sense of agency. Recent developments in self theory emphasize the self as a process rather than a substance or thing. Narrative is the discourse mode most able to express identity as a process. Victimic identity is manifest in a self story in which protagonists have lost power to affect change in their lives; agentic identity is manifest in self-stories of active agency. A study of clients' rehabilitation by Cochran and Laub found that clients' change from victimic to agentic identity moves through four phases: incompleteness, positioning, actualizing, and completion. PMID- 8712250 TI - Jenny's story: reinventing oneself through occupation and narrative configuration. AB - Two life history interviews were conducted to discover how one women, Jenny, experienced a traumatic head injury, rehabilitation, and recovery. Narrative analysis of the transcribed interviews revealed a rich story of how Jenny had fashioned her identity and character through childhood occupations, including studying classical literature and music, and of how she drew upon resources developed in childhood to engineer her recovery. It also illustrated how Jenny used a recursive process of narrative construction and engagement in self-devised graduated occupations, including studying, playing music, writing, computer graphics, and theater production, to create a new identity and develop capacities to process complex information and exercise creativity. Jenny's story illustrates the usefulness of gaining a perspective on patients as occupational beings through the gathering of life histories focused on occupation, the importance of collaborative patient-therapist goal setting, and the necessity for considering both the doing (practic) and the meaning (narrative) aspects of occupation. Her story supports many scholars' arguments that the therapeutic relationship, and thus occupational therapy practice, may be enhanced through the use of life history interviewing in occupational therapy evaluation and treatment. PMID- 8712251 TI - Model demonstrating respiratory mechanics for high school students. AB - We wanted to develop educational materials appropriate for the high school student which would present physiological concepts in an innovative way. The impetus was in response to the lack of physiology educational materials appropriate for the high school level. To this end, we developed an exercise that presents the physiological basis for respiratory mechanics. The materials were designed to engage students in interactive learning and to stimulate interest for future science study. The emphasis of the exercise was the construction of a model that could be built by high school students to demonstrate respiratory mechanics. The use of models to present complex materials has been shown to be an effective medium for science learning. Our exercise contains directions for an inexpensive, easy-to-build model, as well as many supplemental teaching tools. Questions are interspersed throughout the text and at the end of the laboratory experience to facilitate the learning process. Answers are provided to the questions. Students and teachers alike are challenged to build, manipulate, and discuss their experience during the investigation of respiratory mechanics. PMID- 8712252 TI - Directed case study method for teaching human anatomy and physiology. AB - A mastery of human anatomy and physiology requires a familiarity with a vast number of details about the human body. A directed method of case analysis is described that helps students deepen and solidify their understanding of anatomical and physiological facts, concepts, and principles. The successful case had four distinctive features as follows: clear learning objectives, a concise and informative scenario, straightforward and didactic questions, and an emphasis on information readily available to the student. A directed case study is presented, and its salient features are described. A procedure for integrating case analyses into an undergraduate anatomy and physiology course is outlined. Student response to this type of case study suggests that this method improves the ease of learning, the depth of learning, and an appreciation of the relevance of and a curiosity about anatomy and physiology. The addition of case analyses to a two-semester integrated course in anatomy and physiology was also associated with an improvement in exam performance. The regular use of directed case analysis is a valuable addition to the traditional methods of lecture, textbook reading, and laboratory for the teaching of human anatomy and physiology. PMID- 8712253 TI - Muscle chamber with strain gauge adapted for isotonic/isometric recording. AB - Simple devices are described that can facilitate student experiments on mechanical and electrical aspects of muscle contraction, for example, of isolated preparations such as frog gastrocnemius and sartorius muscles and gut. The system includes a general-purpose preparation bath that was previously described for electrophysiological observations with suction electrodes [D.M. Easton. Am.J. Physiol. 265 (Adv. Physiol.Educ. 10): S35-S46, 1993]. A tower installed in the bath allows the muscle to be oriented vertically but completely submerged in a limited volume. Recording of isotonic contraction, which is the more familiar behavior of skeletal muscle, is made convenient by an accessory to convert the commonly available Statham force transducer to isometric recording. Phenomena not conveniently accessible by isometric recording, such as muscle work, and also the length-tension relation, not ordinarily observed for isotonic contraction, can be demonstrated. PMID- 8712254 TI - Graphic format for teaching long-term control of systemic arterial pressure. AB - Circulatory homeostasis is a difficult notion. The graphic format presented here facilitates the teaching of long-term control of systemic arterial blood pressure and cardiac output. It is based on the view that the following four "function curves" cooperate in long-term regulation: the relation between blood volume and ventricular filling pressure, the relation between ventricular filling pressure and cardiac output, the relation between cardiac output and peripheral resistance, and the relation between arterial pressure and natriuresis. Positioning the function curves in the format presented here clarifies their cooperativity. The distinction between a nonsteady state and a steady state deserves emphasis. Long-term pathophysiology of the circulation is most easily taught on the basis of the assumption that, generally, there will be a steady state. The format clarifies why some known physiological relations are almost impossible to demonstrate in the intact organism, and it discourages explanations of pathophysiology that are not firmly based on physiology. PMID- 8712255 TI - A spinal circuitry simulator as a teaching tool for neuromuscular physiology. AB - Many concepts in neuromuscular physiology can be difficult for instructors to teach and for students to understand. The behaviors of various components in neuromuscular systems do not always interact in obvious ways, and the function of hundreds of components can be very different from the function of just one or two "representatives." In this paper, a simulator is presented that can model both small and large spinal circuitry systems thus allowing students to explore the dynamic functional implications of the static circuitry diagrams that are common in many neuroscience textbooks. The simulator brings to life many concepts in neuromuscular physiology and permits students to explore such concepts without extensive supervision. The benefits and drawbacks of using this kind of simulator in the classroom are discussed, based on initial field tests with undergraduate and graduate students as well as input from the literature. It was found that such a simulation can be very useful as a teaching tool if it is used properly with the right audience. PMID- 8712256 TI - Preparing students to participate in an active learning environment. AB - Most students have spent the majority of their school career in passive learning environments in which faculty were disseminators of information, and students were required to memorize information or use specified algorithms to "solve problems." In an active learning environment, students are encouraged to engage in the process of building and testing their own mental models from information that they are acquiring. In such a learner-centered environment, faculty become facilitators of learning, and students become active participants, engaging in a dialogue with their colleagues and with the instructor. To create a successful active learning environment, both faculty and students must make adjustments to what has been their respective "traditional" roles in the classroom. For the instructor who is committed to promoting active learning, the challenge lies in helping students understand the necessity of becoming active colleagues in learning. This process can be facilitated if the curriculum includes exercises to direct students' attention to a number of issues that impact their learning. This paper describes four such exercises designed to help students form appropriate course expectations, recognize the need for seeking clarification when communicating, recognize the role of personal experience in building mental models, and become familiar with study aids for building formal models. PMID- 8712257 TI - Teaching nerve conduction to undergraduates: the "traveling flame" analogy revisited. AB - A familiar analogy to the propagation of the electric wave in a nerve lies in the burning of a cigarette. This analogy, which has persisted through several editions of popular textbooks of physiology, has never been elaborated beyond a cursory mention. In this article, the analogy is reinvoked and refined to explain the cable properties of nerve, saltatory conduction, and some other factors affecting nerve conduction velocity. Experience in lecture classes and feedback obtained through class tests indicate that the analogy, if suitably elaborated and judiciously applied, can increase the retention of the subject by the students. PMID- 8712258 TI - Graphic analysis for the study of metabolic states. AB - Graphic analyses have been used in the study of physiology as a means to better understand dynamic processes and to visualize the mechanisms of their interactions. A graphic analysis of glucose homeostasis was constructed by considering the main factors that influence glucose dynamics. The analysis is achieved by equating curves representing both the inflow and outflow of glucose from the circulation as dependent upon the serum insulin concentration. The point where these two curves intersect is the steady-state balance for blood glucose exchange and is termed the equilibrium point. With the use of this graphic depiction of glucose homeostasis, it is now possible to study the influence of multiple factors on glucose dynamics. A variety of metabolic states can also be analyzed by reconstructing the effects of the pathophysiology on the form and shape of the curves. Some of the metabolic states that have been analyzed by this technique include starvation, exercise, obesity, type I and type II diabetes mellitus, stress, hypopituitarism, hyperpituitarism, and hyperthyroidism. Although the analyses do not reflect all of the controversial nuances of the field, they do provide a means for a general approach to the study of glucose homeostasis and serve as a methodology that can be extrapolated to many areas of physiological study. PMID- 8712259 TI - Tuberculosis control in resource-poor settings with high rates of HIV infection. PMID- 8712260 TI - Revealing and reversing tobacco industry strategies. PMID- 8712261 TI - Great expectations, or do we ask too much from community-level interventions? PMID- 8712262 TI - Why foodborne disease surveillance is critical to the safety of our food supply. PMID- 8712263 TI - Needle exchange programs and the law--time for a change. PMID- 8712264 TI - The role of employee flexible spending accounts in health care financing. AB - Employee flexible spending accounts for health care represent one component of the current health care financing system that merits serious reform. These accounts create a system of undesirable incentives, force employees and employers to take complicated gambles, reduce tax revenues, and fail to meet their purported policy objectives. This paper describes shortcomings in these accounts from both a theoretical and an empirical perspective. Some proposed alternatives; including medical spending accounts and zero balance accounts, resolve many of these concerns but not all of them. PMID- 8712265 TI - Childhood sexual abuse and bulimic behavior in a nationally representative sample. AB - OBJECTIVES: This study examines whether childhood sexual abuse is a significant risk factor for the development of bulimic behavior in women. METHODS: Interviews were conducted among a representative national sample of 1099 US women regarding their sexual experiences during childhood as well as their lifetime prevalence of bulimic behavior. Logistic regression was used to calculate adjusted odds ratios to measure the contribution of childhood sexual abuse to the odds of several measures of bulimic behavior in the population, controlling for the respondent's age, ethnic group, and parents' educational level. Attributable risk analyses were conducted to estimate how much risk for bulimic behavior was due to childhood sexual abuse. RESULTS: Victims of childhood sexual abuse were significantly more likely to display bulimic behavior, and a substantial fraction of cases of bulimic behavior could be attributed to such abuse. CONCLUSIONS: Childhood sexual abuse is a risk factor for bulimic behavior. Between one sixth and one third of the cases of significant bulimic disturbance could be attributed to childhood sexual abuse. PMID- 8712266 TI - Mortality after the death of a spouse: rates and causes of death in a large Finnish cohort. AB - OBJECTIVES: This study examines excess mortality among Finnish persons after the death of a spouse, by sex, the subject's cause of death, duration of bereavement, and age. METHODS: The subjects were 1580000 married Finnish persons aged 35 through 84 years who were followed up from 1986 through 1991. RESULTS: Excess mortality among the bereaved was high from accidental, violent, and alcohol related causes (50% to 150%), moderate for chronic ischemic heart disease and lung cancer (20% to 35%), and small for other causes (5% to 15%). Excess mortality was greater at short ( < 6 months) rather than long durations of bereavement and among younger rather than older bereaved persons for most causes of death; it was also greater among men that women. CONCLUSIONS: The results are consistent with the hypothesis that excess mortality after the death of a spouse is partly caused by stress. The loss of social support or the inability to cope with stress may explain why men suffer from bereavement more than do women. PMID- 8712267 TI - Directly observed therapy for tuberculosis in rural South Africa, 1991 through 1994. AB - OBJECTIVES: This paper describes an audit of a community-based tuberculosis treatment program involving directly observed therapy in South Africa. METHODS: A program audit of 2473 consecutive tuberculosis patients in Hlabisa Health District, KwaZulu/Natal, South Africa, was conducted between 1991 and 1994. RESULTS: Monthly admissions increased from 34 per month in 1991 to 66 in 1994. Of 2186 patients managed in Hlabisa, 1903 (87%) received directly observed therapy. Of those receiving directly observed therapy, 1034 (55%) were supervised by volunteers; 743 (72%) of these were supervised by storekeepers. Among those patients managed locally, 1679 (85%) of 1967 surviving patients completed treatment. Completion rates for patients supervised by health workers and non health workers were the same. Completion fell from a high of 90% in 1992 to 78% in 1994. Mortality increased from 5% in 1991 to 10% in 1994. CONCLUSIONS: Community-based directly observed therapy that uses an intermittent drug regime and volunteers as supervisors can achieve high treatment completion rates for tuberculosis, even in resource-poor settings. PMID- 8712268 TI - Gender differences in sexual practices and sexually transmitted infections among adults in Lima, Peru. AB - OBJECTIVES: This study examined the prevalences of antibodies to Treponema pallidum, Chlamydia trachomatis, and herpes simplex virus type 2 in a sample of Peruvian adults. METHODS: Among adults seeking health certification in Lima, Peru, 600 were randomly selected to undergo interviews and serologic testing. RESULTS: Men's reported mean lifetime number of partners (10.6) far exceeded women's (1.1), yet antibody to sexually transmitted infection pathogens among sexually experienced participants was 2.8 times more prevalent among women than among men. Among men, female sex workers accounted for 37% of recent partners, and only sex with female sex workers while using condoms less than half of the time was independently associated with antibody (odds ratio = 3.6, 95% confidence interval = 1.5, 8.8). among women, number of partners was associated with any sexually transmitted infection antibody, while intercourse before 18 years of age was associated with C trachomatis antibody. At every level of perceived risk, sexually transmitted infection antibody was more frequent among women. CONCLUSIONS: Men having unprotected sex with female sex workers had the greatest risk of acquiring infections and (by inference) of transmitting them to women. PMID- 8712269 TI - Is genital mycosis associated with HIV risk behaviors among heterosexuals? ACSF Investigators. Analyse des comportements sexuels en France. AB - OBJECTIVES: This study examined in both sexes whether genital mycosis is as strongly associated with human immunodeficiency virus (HIV) sexual risk behavior as are other sexually transmitted diseases. METHODS: Heterosexuals in the French National Survey of Sexual Behavior who reported a history of either mycosis or another sexually transmitted disease in the previous 5 years were compared with heterosexuals who reported no such disease. Odds ratios were adjusted for age, education, place of residence, and number of sexual partners. RESULTS: Among men, mycosis and other sexually transmitted diseases showed similar associations with age and number of sexual partners. Among women, mycosis was also associated with a high number of sexual partners, albeit to a lesser extent than other sexually transmitted diseases. Lifetime experience of heterosexual anal penetration was reported significantly more often by men with a history of mycosis. CONCLUSION: These results suggest that most male mycosis is sexually acquired and constitutes a self-reported indicator as good as other sexually transmitted diseases for high risk. Female mycosis may help define an intermediate HIV risk group. PMID- 8712270 TI - A prospective study of syphilis and HIV infection among injection drug users receiving methadone in the Bronx, NY. AB - OBJECTIVES: The purpose of this study was to assess the relationship between syphilis and human immunodeficiency virus (HIV) infection in injection drug users. METHODS: A 6-year prospective study of 790 injection drug users receiving methadone maintenance treatment in the Bronx, NY, was conducted. RESULTS: Sixteen percent (4/25) of HIV-seroconverting patients, 4.8% (16/335) of prevalent HIV seropositive patients, and 3.5% (15/430) of persistently HIV-seronegative patients was diagnosed with syphilis. Incidence rates for early syphilis (cases per 1000 person-years) were 15.9 for HIV-seroconverting patients, 8.9 for prevalent HIV-seropositive patients, and 2.9 for persistently HIV-seronegative patients. Early syphilis incidence was higher among women than men (8.4 vs 3.2 cases per 1000 person-years). Independent risks for early syphilis included multiple sex partners, HIV seroconversion, paid sex, and young age. All HIV seroconverters with syphilis were female. CONCLUSIONS: Diagnosis of syphilis in drug-using women reflects high-risk sexual activity and is associated with acquiring HIV infection. Interventions to reduce the risk of sexually acquired infections are urgently needed among female drug users. PMID- 8712271 TI - HIV infection and AIDS in China, 1985 through 1994. AB - OBJECTIVES: This paper analyzes data on the distribution of and risk factors for the acquired immunodeficiency syndrome (AIDS) and human immunodeficiency virus (HIV) infection in China. METHODS: Ten years of data on persons tested for HIV infection and AIDS and the proportion who tested positive were analyzed against the background of China's population count. The Chinese- and English-language literature on HIV and AIDS from 1985 through 1995 was also reviewed. RESULTS: Overall, more males than females had HIV infection. Intravenous drug use was the primary source of transmission, followed by heterosexual contacts. Only a small number of the persons tested were homosexual, but their proportion of HIV seropositivity ranked third to that of drug users; that of general hospital patients ranked fourth. CONCLUSIONS: HIV infection and AIDS in China began as a highly regionalized and largely rural problem in Yunnan Province. However, HIV infection and AIDS have become an emerging urban problem. HIV seropositivity is low among several groups thought to have an elevated risk. PMID- 8712272 TI - HIV risk behaviors among women living in low-income, inner-city housing developments. AB - OBJECTIVES: This study describes the prevalence and predictors of human immunodeficiency virus (HIV) risk behaviors among women living in low-income, inner-city housing developments. METHODS: Anonymous questionnaires were administered to 671 women living in 10 inner-city, low-income housing developments in five US cities to determine their levels of HIV risk behavior and predictors of HIV risk practices. RESULTS: Approximately one third of women were at high risk for HIV because of the risk behavior of their sexual partners. HIV risk was highest among women who accurately perceived themselves to be at increased HIV risk, held strong beliefs about barriers to condom use, and reported weak behavioral intentions to reduce risk. Women at higher risk were also younger and reported higher rates of alcohol and substance use. CONCLUSIONS: HIV prevention efforts are needed for inner-city women. Interventions should focus on overcoming women's barriers to condom use, strengthening their intentions to change behaviors, and managing the risk related to their use of substances. PMID- 8712273 TI - The Mpowerment Project: a community-level HIV prevention intervention for young gay men. AB - OBJECTIVES: Since young gay men are engaging in alarmingly high rates of unsafe sex and few seek help for changing risky behaviors, community-level programs to prevent infection with the human immunodeficiency virus (HIV) among them are urgently needed. METHODS: We developed and implemented a community-level HIV prevention program in a midsized Oregon community. The peer-led program had three components: out-reach, small groups, and a publicity campaign. Independently from the prevention program, a cohort of young gay men (n = 300) was surveyed in this and in a similar comparison community pre- and postintervention. RESULTS: Following intervention, the proportion of men engaging in any unprotected anal intercourse decreased from 41.0% to 30.0% (-27% from baseline), decreased from 20.2% to 11.1% (-45% from baseline) with nonprimary partners, and decreased from 58.9% to 44.7% (-24% from baseline) with boyfriends. No significant changes occurred in the comparison community over the same period. CONCLUSIONS: This prevention approach effectively led to HIV risk reduction. To reach risk-taking young gay men, HIV prevention activities must be embedded in social activities and community life. PMID- 8712274 TI - State tobacco lobbyists and organizations in the United States: crossed lines. AB - OBJECTIVES: This research took a quantitative look at state-level tobacco lobbying in the United States. METHODS: Publicly available lobbying data were collected from all states during 1994. Data were compiled on tobacco industry lobbyists, their tobacco employers, health lobbyists, and factors associated with such lobbying. RESULTS: In 1994, 450 tobacco industry lobbyists lobbied at a state level. Most lobbying was on behalf of four organizations: Philip Morris (34%), the Tobacco Institute (21%), RJ Reynolds (17%), and the Smokeless Tobacco Council (15%). Approximately one half of all tobacco lobbyists also lobbied for a health-related organization (e.g., state medical association, hospital, physician association). CONCLUSIONS: All US states have tobacco lobbyists. Many health organizations knowingly or unknowingly employ lobbyists who also lobby for the tobacco industry. PMID- 8712275 TI - A new route of transmission for Escherichia coli: infection from dry fermented salami. AB - OBJECTIVES: This study evaluated the production of dry fermented salami associated with an outbreak of Escherichia coli O157.H7 infection in Washington State and California. METHODS: Facility inspections, review of plant monitoring data, food handler interviews, and microbiological testing of salami products were conducted. RESULTS: Production methods complied with federal requirements and industry-developed good manufacturing practices. No evidence suggested that postprocessing contamination occurred. Calculations suggested that the infectious dose was smaller than 50 E. coli O157:H7 bacteria. CONCLUSIONS: Dry fermented salami can serve as a vehicle of transmission for O157:H7 strains. Our investigation and prior laboratory studies suggest that E. coli O157:H7 can survive currently accepted processing methods. PMID- 8712276 TI - Directly observed therapy for tuberculosis: the Harlem Hospital experience, 1993. AB - OBJECTIVES: A directly observed therapy program was established at Harlem Hospital, New York, NY, in 1993 to promote high tuberculosis treatment completion rates. METHODS: The Harlem program used an on-site surrogate family model. Treatment completion rate, visit adherence rate, human immuno-deficiency virus seroprevalence, and time to sputum culture conversion were assessed. RESULTS: Out of 145 enrolled patients with suspected and confirmed tuberculosis, 95 (92 confirmed and 3 suspected) continued treatment. The visit adherence rate was 91.1 +/- 7.9%, with one patient (1%) lost to follow-up. CONCLUSION: High rates of treatment completion and visit adherence were achieved because of unique program characteristics. Thus, directly observed therapy is advocated as a means of ensuring treatment completion. PMID- 8712277 TI - Mass human exposure to rabies in New Hampshire: exposures, treatment, and cost. AB - OBJECTIVES: This study assessed the rabies exposure and treatment that at least 665 persons in Concord, NH, received as a result of one proven rabid pet-store kitten in October 1994. METHODS: All treatment recipients were interviewed by person or phone. RESULTS: The median age of the treatment recipients was 14 years; 58% were female. The most common exposures were low risk (e.g., picking up, petting, nuzzling, or being scratched by a potentially rabid kitten). Local reactions to vaccine or immune globulin were reported by 76.5% of recipients, while 48.8% reported at least one systemic reaction. Cost for the biologicals was estimated at more than $1.1 million. CONCLUSIONS: Because of the inadequacy of pet store records, the inconsistent application of treatment guidelines, and other factors, many people received postexposure treatment as a result of contacts that were unlikely to transmit rabies. The rates of local and systemic adverse reactions experienced were consistent with previous reports. PMID- 8712278 TI - Death rates among patients hospitalized with community-acquired pneumonia: a reexamination with data from three states. AB - OBJECTIVES: Death rates for community-acquired pneumonia based on relatively small-scale, published studies tend to exceed 15% to 20%. This study reexamined these estimates by using very large, population-based databases. METHODS: Death rates from 1993 associated with community-acquired pneumonia were reexamined with hospital discharge data from all of Washington, Illinois, and Florida. RESULTS: These death rates were substantially lower (7.0%, 8.1%, and 9.7%, respectively) than what appears in the literature. Significant risk factors for dying were being 65 years of age or older (odds ratio [OR] = 2.9), being positive for human immunodeficiency virus (OR = 2.9), and having a high severity of illness (OR = 7.1). CONCLUSION: Sampling bias associated with selection for hospital admissions explain the discrepancy between previous and this study's results. PMID- 8712279 TI - A meningococcal vaccination campaign on a university campus: vaccination rates and factors in nonparticipation. AB - OBJECTIVES: This study was undertaken to determine an accurate vaccination rate and identify factors influencing nonvaccination in a meningococcal vaccination campaign on a Connecticut university campus in May 1993. METHODS: Vaccination and student data were merged to determine demographic factors associated with nonvaccination. A case-control study examined reasons for nonvaccination. RESULTS: The estimated vaccination rate for students returning to the campus was 93%. Lower rates occurred among older students, students living off campus, graduate and nondegree students, and married students. Perceived poor access to the vaccination center was the strongest predictor of nonvaccination. CONCLUSIONS: Higher vaccination rates may be achieved by specifically targeting students who live off campus and by providing multiple vaccination sites with extended hours. PMID- 8712280 TI - HIV infection and risk behaviors among male port workers in Santos, Brazil. AB - OBJECTIVES: This paper measured the extent to which human immunodeficiency virus (HIV) infection has spread among the male working-class population of Santos, Brazil. METHODS: Questionnaires on risk behaviors and blood tests were administered to a random sample (n = 395) of male port workers employed by the Santos Port Authority. RESULTS: Although the rate of HIV infection among these men- the working-class male population of Santos-remains low (1.1%), self reported behavioral risks for HIV infection are common. CONCLUSIONS: There is still time to prevent a widespread outbreak of HIV infection among the hetero sexual population of Santos and of the transportation corridors emanating from that city. PMID- 8712281 TI - The legal strategies used in operating syringe exchange programs in the United States. AB - OBJECTIVES: This study sought to identify the strategies used by syringe exchange programs to establish their legality. METHODS: Statutes, court decisions, published studies of exchange programs, and news stories were reviewed, and telephone interviews were conducted with syringe exchange personnel. RESULTS: Twenty-seven exchanges have been authorized by amendments to or judicial interpretations of state drug laws or by administrative action under such laws, or operate in a state that has no laws regulating needles. At least 13 programs operate under claims of legality based on local interpretations of state law, principally public health law. The remaining syringe exchanges operate without a claim of legality. CONCLUSIONS: The deployment of syringe exchanges has been hindered by concerns about their legal status. This study shows that the applicability of drug laws to syringe exchange is open to dispute, and that local public health authorities may under some circumstances rely on their own legal authority to fund or operate syringe exchange programs. PMID- 8712282 TI - Meeting the public's need for information during the Arizona hantavirus outbreak. PMID- 8712283 TI - Diagnosing children with attention deficit disorders through a health department public school partnership. PMID- 8712284 TI - Using volunteers to survey child restraint compliance among New Hampshire drivers. PMID- 8712285 TI - Reducing delays in the laboratory diagnosis of tuberculosis. PMID- 8712286 TI - Questioning the validity of self-reported heterosexual HIV transmission. PMID- 8712287 TI - Specific cutaneous infiltrates of B-cell chronic lymphocytic leukemia: a clinicopathologic and prognostic study of 42 patients. AB - The clinical and histopathologic features of specific skin infiltrates in patients with B-cell chronic lymphocytic leukemia (B-CLL) have rarely been reported in detail. In this study we analyzed the clinical, histopathologic, immunophenotypic, and molecular features of 84 skin lesions from 42 patients (M:F = 1.3:1; mean age, 66.0 years; range, 42-83 years) with specific cutaneous manifestations of B-CLL. The duration of B-CLL before skin manifestations varied from 0 to 142 months (mean, 39 months). In seven patients (16.7%), skin lesions represented the first sign of disease. Clinical presentations included localized or generalized erythematous papules, plaques, nodules, and large tumors. Ulceration was uncommon. In six patients lesions were confined at the sites of scars from previous herpes zoster (four patients) or herpes simplex (two patients) eruptions. Histologically, three main patterns were recognized: (a) patchy perivascular and periadnexal, (b) nodular-diffuse, and (c) band-like. Cytomorphologically, small monomorphous lymphocytes predominated. Proliferation centers were observed in only four specimens. In two patients presenting with tumors, a high content of large cells with feature of centroblasts and immunoblasts was found (Richter's syndrome). Immunohistologic analyses were performed on paraffin-embedded specimens in 40 biopsies from 20 patients and on cryostat sections in 17 biopsies from 11 patients. Neoplastic B lymphocytes in all cases showed an aberrant phenotype (paraffin sections: CD20+/CD5+/CD43+; cryostat sections: CD19+/CD5+; immunoglobulin light-chain restriction). Proliferation markers (Ki67, PCNA, MIB1) stained 5 to 80% of cells (mean, 25%; median, 20%). Polymerase chain reaction performed in nine cases on paraffin embedded tissues using consensus primers for immunoglobulin heavy-chain genes showed a monoclonal population of B lymphocytes in all cases. Several discrete bands in addition to the prominent ones were noted in five cases, indicating the additional presence of B lymphocytes whose immunoglobulin genes were not monoclonally but oligoclonally rearranged. Follow-up data could be obtained from 31 patients. The two patients with Richter's syndrome died after 5 and 8 months, respectively. The 5-year survival of patients with small-cell cutaneous B-CLL was 66.6%. Our study indicates that cutaneous specific manifestations of B-CLL present with characteristic histologic, immunophenotypic, and molecular patterns. Prognosis in these patients is probably not affected by skin involvement. PMID- 8712288 TI - Marginal zone lymphoma (low-grade B-cell lymphoma of mucosa-associated lymphoid tissue type) of skin and subcutaneous tissue: a study of 15 patients. AB - Extranodal low-grade B-cell lymphoma of mucosa-associated lymphoid tissue (MALT) type occurs in the gastrointestinal tract, salivary gland, thyroid, orbit, lung, and breast. We report 15 patients with MALT-type lymphomas involving skin and subcutaneous tissue. All patients had tumors with histologic features of low grade B-cell lymphoma of MALT type, including marginal zone cells (15 of 15 cases), plasmacytic differentiation (10 of 15 cases), Dutcher bodies (three of 15 cases), and reactive germinal centers (10 of 15 cases). All expressed pan B-cell antigens and monotypic immunoglobulin. Seven patients (five women, two men) aged 29 to 86 years (median, 53 years) had primary MALT-type lymphoma of skin (6) or subcutaneous tissue (1). One patient had persistent disease, and four patients had relapses involving skin, subcutaneous tissue, breast, orbit, and lymph node. At last follow-up (11-121 months; median, 36 months), one patient was alive with disease, and six patients had no evidence of disease. Three patients (two women, one man) aged 36 to 67 years (median, 57 years) had concurrent MALT-type lymphoma involving both subcutaneous tissue and extracutaneous sites at primary diagnosis, including lung, breast, orbit, lymph node, and bone marrow. One patient responded to treatment but relapsed with lymphoma of the skin and breast. The other two patients had persistent disease despite treatment. One patient died of disease at 25 months, and, at last follow-up (7 and 46 months), two patients were alive with disease. Five patients (four women and one man) aged 29 to 72 years (median, 63 years) had secondary skin or subcutaneous involvement by MALT-type lymphoma with primary tumors of ocular adnexa (3) or parotid gland (2). All five patients had relapses, which involved skin or subcutaneous tissue, parotid gland, lacrimal gland, breast, and lymph node. At last follow-up (61-137 months), two patients were alive with disease and three were alive with no evidence of disease. Low grade B-cell lymphomas of MALT type may arise in or secondarily involve the skin and subcutaneous tissue and have a tendency to affect middle-aged to older women. These tumors are characterized by multiple extranodal relapses and are associated with long patient survival. Patients with primary MALT-type lymphoma of skin or subcutaneous tissue without extracutaneous involvement at diagnosis were more likely to experience prolonged disease-free survival than patients with extracutaneous spread at presentation (p < 0.03). PMID- 8712290 TI - Significance of interdigitating reticulum cells in Omenn's syndrome. PMID- 8712289 TI - Diffuse colonic mantle cell lymphoma in a patient with presumed ulcerative colitis: detection of a precursor monoclonal lymphoid population using polymerase chain reaction and immunohistochemistry. AB - Primary lymphoma of the colon, a rare and typically late complication of ulcerative colitis, exhibits high-grade morphology and behavior when it occurs. Recently, several reports of colonic lymphoma masquerading as ulcerative colitis have been described. These previous reports described inflammatory mucosal changes typical of ulcerative colitis as being present in superficial biopsies, leading to the initial diagnosis of ulcerative colitis; however, further workup resulted in a diagnosis of primary colonic lymphoma within several months in these cases, and all symptoms and mucosal changes resolved after treatment of the lymphoma. Herein we report a case of mantle cell lymphoma arising in the colon and rectum in a 71-year-old woman with a 4-year history of ulcerative colitis. Immunoglobulin heavy-chain gene rearrangements were detected using the polymerase chain reaction procedure in fixed tissue in the lymphoma as well as in a prior resection specimen that histologically appeared to show only changes of severe ulcerative colitis. This finding suggests that an indolent lymphoid proliferation may have been the underlying disease in this patient and raises questions about the role of colonic lymphoma in causing mucosal injury. PMID- 8712291 TI - Gnathic diagnosis. PMID- 8712292 TI - Focal versus diffuse anaplasia in Wilms tumor--new definitions with prognostic significance: a report from the National Wilms Tumor Study Group. AB - Anaplasia, defined by the presence of extreme nuclear and mitotic atypia, is a potent marker of adverse prognosis in Wilms tumor (WT). Anaplastic WT cells apparently have increased resistance to therapy rather than increased aggressiveness. The distribution of anaplasia should therefore have critical prognostic relevance. The original definitions for focal anaplasia (FA) and diffuse anaplasia (DA) were based on quantitative rather than topographical criteria and lacked prognostic significance. A new definition was developed based on the distribution of anaplastic changes within the tumor: FA applies only to tumors with anaplasia confined to one or a few discrete loci within the primary tumor, with no anaplasia or marked nuclear atypia elsewhere. This revised definition was evaluated in 165 cases with anaplastic WT entered on the third and fourth National Wilms Tumor Study. Only three relapses and one death occurred among 39 cases with FA, regardless of tumor stage, a result comparable to that for nonanaplastic WT. Eight children with metastases at diagnosis and FA in the primary tumor were alive and free of relapse; 22 of 23 children with stage IV DA WT died of tumor. This new definition reinforces the importance of carefully documenting the exact site from which each tumor section is obtained. PMID- 8712293 TI - Endocrine ductal carcinoma in situ (E-DCIS) of the breast: a form of low-grade DCIS with distinctive clinicopathologic and biologic characteristics. AB - Endocrine ductal carcinoma in situ (E-DCIS), first characterized by Cross et al. in 1985, is an uncommon entity, and there is little information on its pathobiologic features and natural history in the literature. This report describes the largest series of 34 cases: 14 cases were pure in situ (group A), and 20 were accompanied by an invasive component (group B). All except three patients were over the age of 60 years, with the mean being 69.5 years for group A and 72.6 years for group B. Except for six patients in group A who had nipple discharge, all had a breast mass. On follow-up, one of five group A patients developed local recurrence 5 years after mastectomy, and two of seven group B patients developed another invasive primary in the contralateral breast. Histologically, E-DCIS showed expansile intraductal growths forming solid sheets and festoons traversed by delicate fibrovascular septa. Accumulation of basophilic mucin might be found within the growth and the fibrovascular septa. There were variable degrees of stromal sclerosis. In some cases, the solid intraductal cellular proliferations were focally punctuated by microglandular spaces and rosettes. Comedo necrosis was absent. Intraductal papillomas were found in the immediate vicinity of the tumors in 18 cases and invariably showed pagetoid involvement by E-DCIS. Pagetoid spread into the adjacent ducts and ductules was also a common feature (17 cases). The tumor cells were polygonal, oval, or spindly, often with eccentrically placed, bland-looking, ovoid nuclei and abundant eosinophilic granular cytoplasm. Intracellular mucin was commonly demonstrable. Immunostaining for myoepithelium using muscle-specific actin antibody confirmed the in situ nature of the E-DCIS component. The majority of tumor cells showed strong staining with the neuroendocrine markers chromogranin, synaptophysin, and neuron-specific enolase (monoclonal). Immunostaining also dramatically highlighted the pagetoid spread into the papillomas and ductules by outlining the tumor cells between the negatively stained residual ductal epithelium and myoepithelium. All cases were immunoreactive for estrogen and progesterone receptor, but not p53 and c-erbB2. The Ki-67 index was < 5%. Ultrastructural studies on four cases showed many dense-core neurosecretory granules and larger mucigen granules. In group B cases, the invasive component, which comprised 5-95% of the tumor, included colloid carcinoma, 12; "carcinoid" tumor, 3; mixed "carcinoid"/colloid carcinoma, 4; and small cell neuroendocrine carcinoma, 1. Neuroendocrine markers were also consistently demonstrable in the invasive component. In conclusion, E-DCIS is predominantly a disease of older women that is frequently accompanied by papillomas in the vicinity and may present as nipple discharge (an uncommon presentation in the usual forms of DCIS). It can mimic epitheliosis histologically, but the pagetoid spread is a helpful clue to its neoplastic nature. The bland nuclear morphology, lack of necrosis, and biologic marker profile suggest that E-DCIS is a form of low-grade DCIS despite its solid growth pattern. The invasive carcinomas associated with E DCIS are also neuroendocrine programmed rather than the usual types of ductal carcinomas, suggesting that E-DCIS represents a biologically distinctive category of DCIS. PMID- 8712294 TI - Follicular dendritic cell tumor: report of 13 additional cases of a distinctive entity. AB - Follicular dendritic cell (FDC) tumor is an extremely rare malignant neoplasm with approximately 17 well-documented cases in the literature. We report 13 additional cases of this distinctive neoplasm. There were seven men and six women, with a mean age of 46.5 years (range, 27-62 years). There was involvement of cervical lymph nodes (six cases), mediastinum (three cases), axilla, tonsil, spleen, and peripancreatic soft tissues (one case each). The neoplasms were grey to tan, ranging in size from 1 to 13 cm. They were formed by oval to spindle cells with eosinophilic cytoplasm growing in sheets and fascicles, with a focal storiform pattern and whorls reminiscent of those seen in meningioma. The nuclei were oval or elongated with thin nuclear membranes, inconspicuous or small eosinophilic nucleoli, and clear or dispersed chromatin. Typically, the tumor cells were intimately admixed with small lymphocytes, with a prominent perivascular cuffing. Multinucleated tumor cells were present in seven cases. Necrosis, marked cellular atypia, high mitotic rate, and/or abnormal mitoses were present in seven cases. The tumor cells were positive for CD21 (10 of 11), CD35 (10 of 11), Ki-M4p (seven of eight) Ki-FDRC1p (six of seven), vimentin (five of nine), and S100 protein (five of nine). One case stained with actin. In situ hybridization, done in six cases, did not show Epstein-Barr virus RNA sequences. Ultrastructural examination of eight cases showed long, complex, occasionally interdigitating cytoplasmic processes joined by desmosomes. The behavior of these tumors is more akin to that of a low-grade soft tissue sarcoma than a malignant lymphoma and is characterized by local recurrences and occasional metastases. Two patients died of tumor, two were alive with recurrent or metastatic disease, eight were alive with no disease, and one was lost to follow-up. PMID- 8712295 TI - Alpha 1-antitrypsin expression in human thyroid papillary carcinoma. AB - Alpha 1-antitrypsin is a plasma serine protease inhibitor originally used as a marker for tumors of histiocytic origin. Our casual finding of immunoreactive alpha 1-antitrypsin in one case of thyroid papillary carcinoma led us to investigate its presence in 10 thyroid papillary carcinomas by applying immunocytochemical and immunochemical techniques to tissue sections and Western blots of tissue homogenates prepared from neoplastic tissue and from uninvolved normal areas in the vicinity of each tumor. The immunocytochemical study was performed in both thyroid tissue and metastatic regional lymph nodes. This analysis revealed immunoreactivity for alpha 1-antitrypsin in nine of the 10 cases studied. Immunoreactivity was intense in some of the cells forming the papillar and follicular structures. These cells were intermingled with completely unstained tumoral cells. In contrast to neoplastic tissue, the normal thyroid tissue present in the vicinity of each tumor showed no staining for alpha 1 antitrypsin. The electrophoretic analysis performed on homogenates prepared from both tumoral and normal thyroid tissue revealed a drastic reduction in the band corresponding to thyroglobulin in the tumoral tissue compared with normal thyroid extracts, where it represented the major protein. Western blotting and immunoprinting with a polyclonal alpha 1-antitrypsin antibody confirmed the results obtained with immunocytochemistry about the presence of this protease inhibitor in neoplastic thyroid tissue. Immunoprinting with the anti-alpha 1 antitrypsin antibody revealed an intense immunoreactive band of 53 kDa in the extracts prepared from tumoral tissue. This band had exactly the same apparent molecular mass previously described by others for alpha 1-antitrypsin purified from plasma and was identical to the molecular mass of the purified commercial standard employed. PMID- 8712296 TI - Tall cell variant of papillary thyroid carcinoma: a reassessment and immunohistochemical study with comparison to the usual type of papillary carcinoma of the thyroid. AB - A review of 92 consecutive cases of papillary thyroid carcinoma diagnosed at The Methodist Hospital revealed 11 tall cell variant (TCV) cases in nine women and two men. There was a greater average age and larger tumor diameter of TCV cases compared with papillary thyroid carcinoma of the usual type (UPTC), but these differences were not statistically significant. Extrathyroidal extension of tumor was noted in nine of 11 TCV cases and was intraoperatively evident in five cases. The presence of extrathyroidal extension represented a statistically significant difference between TCV and UPTC (p = 0.0001) in a multivariate stepwise logistic regression analysis, with controls for variables of age, sex, tumor size, and lymph node metastases. In 11 TCV patients, tumor recurrence was present in two cases, and there was one tumor-associated death with 1 to 4 years of follow-up. Immunohistochemical stains for thyroglobulin, vimentin, keratins, and Leu-7 were positive in all TCV cases and in 16 of 16 UPTC. Immunoreactivity with antibodies to Leu M1 antigen, a myelomonocytic marker included in cluster designation group (CD 15), which is present in many adenocarcinomas, was present diffusely in all TCV, in contrast to UPTC (with sparse immunostaining in only one of 16 cases). Immunoreactivity with antibodies to ZC-23, an anti-carcinoembryonic antigen (CEA) monoclonal antibody with cross-reactivity to nonspecific cross-reacting antigen and biliary glycoprotein antigen, was present in all TCV but was not present in UPTC. COL-1, a CEA-specific monoclonal antibody, was nonimmunoreactive with all TCV and UPTC cases. Epithelial membrane antigen (EMA) was present in all TCV but was also present focally in eight of 16 UPTC, sometimes in a membranous pattern in epithelium surrounding cystic or hemorrhagic spaces. Strong immunoreactivity with antibodies to Leu M1 and EMA in papillary carcinomas of the thyroid has been associated with advanced stages of disease and tumor-associated mortality. The pattern of immunoreactivity in TCV is dissimilar to that in UPTC and is supportive evidence that TCV is a neoplasm that is distinct from papillary thyroid carcinoma of the usual type. PMID- 8712297 TI - Subpubic cartilaginous cyst: report of two cases. AB - We report two cases of an unusual type of subpubic cystic mass occurring in postmenopausal women. They are formed by fibrocartilaginous tissue with extensive cystic degenerative changes and appear to be related to and perhaps originating from the periarticular tissues of the symphysis pubis. We could not find any reference to this lesion either in the standard gynecology and pathology textbooks or in a search of the published literature. PMID- 8712298 TI - Intraductal oncocytic papillary neoplasms of the pancreas. AB - We describe the clinical and pathologic features of 11 intraductal oncocytic papillary neoplasms of the pancreas, a hitherto unrecognized tumor. The patients were six men and five women, and most of the tumors were in the head (head: body/tail = 8:3). The mean patient age was 62 (range, 39-78), and the average tumor size was 6 cm. Grossly the tumors exhibited mucin-filled cysts containing nodular papillary projections. Dilated ducts communicating with the main tumor were sometimes noted. Microscopically the cystic structures appeared to represent dilated ducts containing intraductal tumor. The tumors were characterized by variably complex, arborizing papillary structures. The papillae had thin, delicate fibrovascular cores with focal myxoid changes and were lined by stratified oncocytic cells. Goblet cells and intra-epithelial mucin-containing lumina were present, the latter resulting in a characteristic cribriform pattern. The exuberance of the epithelial proliferation varied from case to case and between different regions within individual tumors; solid sheets of cells were often identified. Although the degree of cytologic atypia was not generally severe, the complexity of the architecture justified a designation of intraductal oncocytic papillary carcinoma in 10 of the 11 cases. In nine cases the tumor was entirely intraductal; one case exhibited focal microinvasion and another showed widespread invasive carcinoma, the invasive elements appearing cytologically similar to the intraductal papillary components. The oncocytic cells stained positively with phosphotungstic acid hematoxylin and Novelli stains. Immunohistochemically, all cases stained positively for B72.3, and five cases showed focal, weak luminal membrane staining for carcinoembryonic antigen. Ultrastructurally many of the cells were packed with mitochondria, and mucin was also identified. Seven patients were alive and free of tumor from 1 month to 3 years (average, 1 year) after resection. Two patients died postoperatively. The remaining two patients died with no evidence of disease at 2.5 and 5 years, the latter following a recurrence at 2.5 years. We conclude that intraductal oncocytic papillary neoplasm is a distinctive pancreatic tumor that is usually intraductal but may develop invasive carcinoma and should be treated with complete resection. PMID- 8712299 TI - Pouch-anal cancer after restorative proctocolectomy for familial adenomatous polyposis. AB - Familial adenomatous polyposis (FAP) and idiopathic ulcerative colitis are both mucosal diseases that bear a significant risk of developing colorectal cancer. As a consequence, their surgical treatment is currently widely performed by restorative proctocolectomy with pelvic ileal pouch-anal anastomosis (IPAA). Herein we report a new case of cancer after IPAA that developed in a 33-year-old woman with FAP. Pouch-anal cancer was diagnosed 8 years after restorative proctocolectomy, including anorectal mucosectomy, for FAP with manifest cancer in the sigmoid colon. This case observation reemphasizes that the risk of cancer for FAP patients persists as long as any rectal mucosa remains after IPAA surgery. PMID- 8712300 TI - A population-based study of paralytic shell fish poisoning in Alaska. AB - During May and June 1994, the authors interviewed and requested a shellfish sample from a population-based sample of 170 residents from Kodiak and Old Harbor, Alaska. Of 51 Old Harbor and 68 Kodiak residents who had eaten shellfish gathered from Kodiak Island, 18 and 6 percent, respectively, had a history of PSP. We calculated the incidence of paralytic shellfish poisoning in Old Harbor and Kodiak as 15 and 1.5 per 1000 persons per year, respectively. Of 12 butter clam batches collected from residents, 6 had a paralytic shellfish poison toxin level greater than the regulatory limit of 80 micrograms saxitoxin equivalent per 100 g of tissue; one of the 29 people who ate these shellfish developed illness. People who eat shellfish collected from non-commercial beaches have a high rate of paralytic shellfish poisoning. It may be possible to raise the regulatory level for paralytic shellfish poison toxin without affecting the public health. PMID- 8712301 TI - Tribal-based cancer control activities among Alaska Natives: services and perceptions. AB - Cancer has become a significant health problem in American Indian and Alaskan Native (AI/AN) communities. Despite the precipitous rise in cancer rates, limited data are available concerning cancer control services operative in these communities. To address this issue, a cross-sectional survey of all federally recognized tribes was undertaken to ascertain the breadth of cancer control activities offered and Tribal Health Directors perceptions of and priorities ascribed to cancer. This article presents responses given by AN Health Directors juxtaposed to those proffered by AI Health Directors. Nearly three-quarters (71%) of respondents perceived cancer rates to be increasing. Cancer was found to rank third, fifth among AI Health Directors, among seven health conditions when Directors were asked to rank their Tribe's commitment to confronting each one. Awareness of cancer as a public health concern coupled with competing health problems relegates cancer control activities to a lower priority. Findings underscore the need to elevate the issue of cancer in Indian Country as well as to educate investigators to become more sensitive and responsive to other Tribal health issues. PMID- 8712302 TI - Smoking cessation products and programs. AB - Behavioral treatment techniques have facilitated smoking cessation, with intensive multicomponent interventions sometimes producing long-term abstinence rates approaching 50%. There is little evidence that either hypnosis or acupuncture are effective. Both nicotine gum and nicotine patch significantly improve treatment outcomes, although patch is easier for patients to use correctly. Self-help programs may be of benefit, however, smoking cessation products other than nicotine replacement show little evidence of effectiveness. Health professionals should be informed consumers and should be skeptical in evaluating claims for commercial programs or products. Additional information and materials are available from a number of sources including the National Cancer Institute, the U.S. Office on Smoking and Health, and the voluntary health organizations. PMID- 8712303 TI - Prevention of ovarian cancer with bilateral oophorectomy at time of hysterectomy. AB - A review of all patients with a diagnosis of ovarian cancer for five years (1989 1993) in the military hospital, the public health hospital and the two private hospitals in Anchorage, Alaska was conducted. Fifty-eight women were found with a diagnosis of advanced ovarian cancer. These charts were reviewed to determine the incidence of prior hysterectomy with preservation of ovaries. Of the fifty-eight, thirteen (22%) had prior hysterectomies with the ovaries left in place. Review of the literature supported this statistic in similar studies. Ovarian cancer has such a poor prognosis that prevention is in order. The data demonstrate that ovarian cancer could have been prevented in a significant number of these patients had they had bilateral oophorectomies at the time of hysterectomy. With the relative safety of hormonal replacement therapy, a significant way to prevent ovarian cancer is available--bilateral oophorectomy. PMID- 8712304 TI - A survey of the practice of prophylactic oophorectomy by gynecologists in the state of Alaska. AB - A questionnaire was sent to Alaskan gynecologists to evaluate their practice in regard to prophylactic oophorectomy at the time of hysterectomy. At issue is the possible benefit of prophylactic oophorectomy in the prevention of ovarian cancer. Sixty-seven questionnaires were sent and 42 (63%) were returned completed. The percentage who said they would remove apparently normal ovaries at the time of hysterectomy in premenopausal women is as follows: at 35-39 years, 5%; 40-44, 35%; 45-49, 81% and over 49 years, 95%. In postmenopausal women, 97% would recommend removal of the ovaries. 29% of respondents believe that more than 10% of ovarian cancers could be prevented by prophylactic oophorectomy at the time of hysterectomy. PMID- 8712305 TI - Sexually speaking .... From lover to caregiver.... A series of the effect of illness on intimacy. Part one. PMID- 8712306 TI - Acquired corneal asigmatism. PMID- 8712307 TI - [Jugular phlebectasia: a report of two cases]. AB - We report 2 cases of internal jugular phlebectasia addressed to our ENT Department for investigation. The sole clinical symptomatology was a latero cervical right mass expanding with Valsalva's manoeuvre. We make an actualization of the theme's literature and recommend the echography as a method of choice for diagnose, being that technique non-invasive, quick and reproduceable. PMID- 8712308 TI - [Acute pulmonary edema as complication of adenotonsillectomy: a contribution of two cases]. AB - Pulmonary edema secondary to adenotonsillectomy is a serious surgical problem occurring in association with previous chronic, long-lasting obstructive complaints of the upper airway. We report 2 cases in the immediate postoperative. Discussion of its physiopathology. PMID- 8712309 TI - [Diffuse sclerosing chronic osteomyelitis of the mandible]. AB - Diffuse sclerosing chronic osteomyelitis is a lasting pathological entity characterized for recurrent outbreaks of intensive pain referred to the mandible, escorted with inflammatory phenomena, trismus, paresthesia of the nervus alveolaris inferior and progressive deformity of the lower jaw. The differential diagnosis with other facial neuralgies of the face must be very carefully get up and its treatment is disappointing. The etio-pathology of the illness remains uncertain. The AA. report one clinic case with typical course, which offer them the opportunity to review and discuss the literature of the subject. PMID- 8712310 TI - [Primary nasal tuberculosis]. AB - Nasal tuberculosis is a rare entity since it early diagnosis and the improvements in treatment. The mucous membrane of the nose is scanty auspicious for this pathology in spite of being an important entrance door for Koch's bacillus. We report one case of nasal tuberculosis without other primary focal point and emphasize the importance of biopsy and the histo-pathological study in order to reach the positive diagnosis and the healing "ad integrum" by means of suitable antibiotic drugs. PMID- 8712311 TI - [Esophageal perforations by foreign bodies. A contribution of three cases]. AB - Oesophageal perforations due to foreign bodies are a rare pathology with an increased index of complications. Early diagnosis and treatment can reduce the mortality, which accounts for 28 percent. We report 3 cases of oesophageal perforations due to foreign bodies, 2 of them localized at the cervical level and the 3rd, at the end of the thoracic oesophagus, the later ending ominously. PMID- 8712312 TI - [Sjogren syndrome: clinics and ENT diagnosis]. AB - Prospective study covering 38 cases of Sjogren's syndrome (S.S.) diagnosed after early clinical suspicion awaken for xerostomy and xerophtalmy, once excluded those cases which pathology was previously known. Thirty-two cases pertaining to primary malady (P.S.S.) being secondary (S.S.S.) the others. The clinical E.N.T. symptomatology stand out in our series, because the throat dryness and taste and olfactory alterations. The 53 percent of patients have consulted with other specialists without actualizing the diagnosis. The AA. discuss complementary diagnostic methods (Schirmer's test, rose bengal stain, sialometry, auto antibodies and biopsy of minor salivary glands) and diagnostic criteria, and fix the otolaryngologist role in the management of these patients. PMID- 8712313 TI - [Nasopharyngeal melanoma: a case report]. AB - The AA. present a case of disseminated primitive melanoma of the rhinopharynx. The main difficulty was to establish the differential diagnosis, owing to the rarity of this tumor's type in such a sitting. The patient showed intracranial metastases which took him to death in a short term. PMID- 8712314 TI - [Anatomical alterations of the upper airway in the sleeping apnea syndrome. Diagnostical and therapeutic implications]. AB - In this paper are assessed through ENT-exam, rhinomanometry and cephalometry both bipedal and in dorsal decubitus, the anatomical alterations occurring in the upper airways in 100 consecutive patients, polysomnographically diagnosed of SAOS. The patients were classified in 4 groups (after Fujita's modified classification): 2 having oropharyngeal obstruction (I and IIa) and other 2 with hypopharyngeal obstruction (IIb and III), which were subdivided in its time with nasal obstruction present (N+) or absent (N-). Group I and IIa (68 %) theoretical candidates to an uvulopalatopharyngoplasty, UPPP-showed its predominance, followed by a high incidence of nasal obstruction (N+, 48 %) and a non negligeable number of hypopharyngeal alterations (32 %). PMID- 8712315 TI - [Pseudocyst of the auricle]. AB - Reporting one case of pseudocyst of the auricle in a young male, 18, white and wealthy, diagnosed and treated. Bibliographical review of this condition and explanation of its etiopathogenetic, clinic and therapeutical characteristics, as well. This deformity seems to be more prevalent as the number of published cases may suggest. PMID- 8712316 TI - [Mucocele of the submaxillary salivary gland]. AB - Paper reporting a rare case of mucocele of the right submaxillary gland in a youngster, 14 year-old. Perusal of the concerning literature and analysis of its etiopathological theories, the differential diagnosis and comments on diagnostic and therapeutic data as well. PMID- 8712317 TI - Trainee representation in anaesthesia--past, present and future. PMID- 8712318 TI - The effect on the heart of hypoxaemia in patients with severe coronary artery disease. AB - The aim of the study was to examine the effect of spontaneous nocturnal hypoxaemia on myocardial ischaemia in patients with severe coronary artery disease. This was a prospective study of continuously measured oxygen saturation and ECG overnight in patients prior to coronary artery bypass graft surgery. Fifteen patients with angiographically proved coronary artery disease were studied between October 1992 and September 1993. All patients had episodes of hypoxaemia with oxygen saturation < 94% and eight of the 15 patients had episodes where oxygen saturation < 85%. An episode was defined as a period of hypoxaemia not less than 2.5 min in duration, the longest total duration of hypoxaemia in one patient being 355 min. Twelve of the 15 patients showed a drop in ST segment of 1 mm or more lasting from 5-700 min. The hypoxaemic and ischaemic episodes apparently occurred at random. No causal relationship could be shown between hypoxaemia and ischaemia. PMID- 8712319 TI - Anaesthesia and the QT interval in humans. The effects of isoflurane and halothane. AB - Prolongation of the QT interval may cause potentially hazardous arrhythmias. The effects on the QT interval (QTc, corrected for heart rate) of isoflurane and halothane followed by vecuronium have been investigated during induction of anaesthesia in 51 patients. All patients were ASA 1 or 2, without cardiovascular problems or electrolyte abnormalities and were not receiving medication. Midazolam 0.08 mg.kg-1 was administered intramuscularly for premedication. Anaesthesia was induced with either isoflurane (n = 26) or halothane (n = 25), and the inspired concentration increased to reach an end-tidal concentration of 2.5% to 3%. Recordings of ECG, heart rate, systolic and diastolic arterial pressure were obtained at the following times: prior to induction of anaesthesia; 1 min and 3 min after a stable end-tidal concentration had been reached; 1 min and 3 min following vecuronium administration, at the time of tracheal intubation and 1 min and 3 min later. Halothane significantly shortened QTc (p < 0.05 to p < 0.001), in contrast to isoflurane which prolonged it (p < 0.01). The heart rate decreased (p < 0.01 to p < 0.001) after induction of anaesthesia with halothane and returned to pre-induction values after tracheal intubation. In contrast, heart rate increased after induction with isoflurane and increased further after laryngoscopy and tracheal intubation (p < 0.001). In the isoflurane group, ST depression was noticed in seven patients and nodal rhythm in two, while in the halothane group seven patients developed nodal rhythm and, in two patients, ventricular ectopics were recorded. There were no sequelae. In both groups, systolic and diastolic arterial pressure decreased after induction of anaesthesia (p < 0.01 to p < 0.001), increasing again after intubation. PMID- 8712320 TI - The effect of posture on late postoperative oxygenation. AB - Twelve patients had their arterial oxygen saturation measured pre-operatively and on days 1, 4 and 7 after laparotomy. Measurements were performed in the supine, sitting and standing positions on each day. Arterial oxygen saturation was significantly higher during sitting and standing on days 1 and 4 after operation compared with the supine position (p < 0.05). These results give further evidence for the benefits of patient mobilisation after major surgery. PMID- 8712321 TI - National provision of acute pain services. AB - In order to determine the degree to which the recommendations of the report of the joint working party on 'Pain after Surgery' by the Royal College of Surgeons of England and the College of Anaesthetists have been implemented, a postal survey was conducted of all hospitals in the United Kingdom where surgery is performed. The number of hospitals with a multidisciplinary acute pain service had significantly increased from 2.8% before September 1990 to 42.7% at the end of 1994. Over the same period there has been an increase in the number of hospitals with a named clinician responsible for acute pain management, from 20 before 1990 to 230 (65.2%) and the number of hospitals with an acute pain nurse has increased from 8 (2.3%) prior to 1990 to 139 (39.3%). Routine assessment of pain and sedation occurs in 82% of hospitals with established acute pain services and in less than 50% of hospitals without an acute pain service. The use of written protocols, the provision of out-of-hours cover and regular training for all staff have increased with time. Research and audit activity related to acute pain management has also improved since 1990. PMID- 8712322 TI - Tropisetron and metoclopramide in the prevention of postoperative nausea and vomiting. A comparative, placebo controlled study in patients undergoing ophthalmic surgery. AB - One hundred and twenty patients undergoing elective ophthalmic surgery under general anaesthesia were investigated in a randomised, double-blind, parallel group study of postoperative nausea and vomiting. Patients received tropisetron 0.1 mg.kg-1, metoclopramide 0.25 mg.kg-1 or placebo given at the end of anaesthesia. In comparison with placebo, tropisetron significantly reduced the degree of nausea (p < 0.01), whereas metoclopramide reduced both nausea (p < 0.05) and vomiting (p < 0.05). There were no statistically significant differences between the two active agents in their efficacy to postoperative nausea and vomiting. The patients in the placebo group required rescue antiemesis more often in the postanaesthesia care unit. Our results suggest that tropisetron may not be suitable as a routine, primary therapy for the prevention of postoperative nausea and vomiting. PMID- 8712323 TI - The influence of cold on the recovery of three neuromuscular blocking agents in man. AB - The Arrhenius hypothesis suggests that change in temperature has a less marked effect on the rate of physical processes than on biological reactions. We have investigated the process underlying recovery from neuromuscular block in man by studying the effect of cooling on the rate of recovery from depolarising and non depolarising block. Vecuronium, rocuronium and decamethonium (C10) neuromuscular block were investigated using the isolated forearm technique on awake human volunteers. In these experiments, one arm was cooled whilst the other was used as control. Moderate hypothermia decreased the rate of recovery from all three agents, but this was significantly less marked with the depolarising drug. The mean Q10 (the anticipated change in rate of a reaction across of 10 degrees C temperature gradient) of the rate of recovery for vecuronium was 3.21, rocuronium 2.86 and decamethonium 1.29. This suggests a different process in the recovery of these two types of drug. According to the Arrhenius hypothesis this would suggest that the recovery from non-depolarising drugs is likely to involve a biochemical mechanism and that recovery from decamethonium is controlled by a physical process. PMID- 8712324 TI - Measuring rebreathing in spontaneous ventilation. A new lung model. AB - The performance of breathing systems is tested by means of physical lung models. We describe a new lung model, allowing direct measurement of the dead space/tidal volume ratios produced by any breathing system at different fresh gas flows. The model allows a range of different patterns of spontaneous respiration to be studied. Although rebreathing is measured by capnography, the design ensures that the results are unaffected by changes in carbon dioxide inflow, or even calibration drift in the capnograph. PMID- 8712325 TI - Circle systems with a coaxial inspiratory limb. Investigation with a lung model. AB - A laboratory study of a new circle system is presented. The inspiratory limb encloses a coaxial tube, delivering fresh gas close to the patient, immediately upstream of the inspiratory one-way valve. The system is more efficient than the non-coaxial system and allows the expiratory valve to be situated conveniently at the canister end of the expiratory limb. A disadvantage of low flow anaesthesia is the sluggish response of the system to changes in vapour or oxygen concentrations in the fresh gas flow. This is markedly improved by the new system. The advantages of the system are largely independent of respiratory pattern. PMID- 8712326 TI - Minimising cardiac anaesthetic risk: the tortoise or the hare? AB - There is no doubt that a group of patients at increased risk of peri-operative cardiac morbidity exists and must be managed with the emphasis on the prevention of myocardial ischaemia. It is also clear that a potentially far larger group are at risk of failing to meet the increased cardiovascular and metabolic demands of surgery and therefore suffering the consequences of a relative hypoperfusion injury. Pre-operative assessment must address both groups and management regimens sought to provide optimal outcome for both. At present there is no consistent strategy for their identification, assessment or management of the high risk surgical population despite the fact that they probably consume a disproportionate share of hospital resources. The first and most important step is the recognition that this high risk group exists. Only then can this population be given similar consideration to those currently thought to be at risk of ischaemia. PMID- 8712327 TI - Delayed paraplegia following spinal anaesthesia. Spinal subdural haematoma following dural puncture with a 25 G pencil point needle at T12-L1 in a patient taking aspirin. AB - We report an extremely rare complication of regional anaesthesia, a spinal subdural haematoma, which resulted in permanent neurological damage occurring 8 days after dural puncture at T12-L1. Although spinal subdural haematoma following spinal anaesthesia and lumbar puncture has been described before, this is the first report of this complication occurring after dural puncture using a 25 G atraumatic pencil point (Whitacre) needle. Contributory factors might have been the perioperative intermittent low dose aspirin therapy and the fact that spinal anaesthesia was performed at the T12-L1 level. PMID- 8712328 TI - Epidural infusion of low-dose bupivacaine and opioid in labour. Does reducing motor block increase the spontaneous delivery rate? AB - Labouring women were randomly allocated to receive epidural infusions during labour of either 0.125% plain bupivacaine (n = 200) or a combination of 0.0625% bupivacaine with either 2.5 micrograms.ml-1 fentanyl or 0.25 micrograms.ml-1 sufentanil (n = 199) each starting at 12 ml.h-1 and adjusted as necessary to maintain analgesia. The dose of bupivacaine, both hourly (p < 0.001) and total (p < 0.001), was significantly lower in the group receiving the combination. Motor block was significantly less common and less severe in the combination group (p < 0.001). These reductions did not result in a significant increase in spontaneous deliveries. Maternal satisfaction with first (p < 0.001) and second stage analgesia (p < 0.001) was significantly increased in the combination group. The addition of opioid to the epidural infusion did not reduce the incidence of perineal pain. There were no significant differences between the groups in neonatal outcome or the incidence of early postnatal symptoms. PMID- 8712329 TI - The effect of increasing the dose of buprenorphine on the haemodynamic response associated with tracheal intubation. AB - The effect of buprenorphine on the haemodynamic response to tracheal intubation was studied at two dose levels, 2.5 micrograms.kg-1 and 5 micrograms.kg-1, in a placebo-controlled double-blind trial in 75 patients undergoing laparoscopic cholecystectomy. The study drugs were administered intravenously 8 min before induction of anaesthesia with thiopentone 5 mg.kg-1 and vecuronium 0.1 mg/kg-1. Buprenorphine 2.5 micrograms.kg-1 caused 50% attenuation of the blood pressure response whereas 5 micrograms/kg-1 caused 70% attenuation compared to the saline placebo. The maximum increase in heart rate was 14% of the control value after 2.5 micrograms.kg-1 and 11% after 5 micrograms/kg-1 of buprenorphine. A significant difference in heart rate was also observed between the two buprenorphine groups at 5 and 10 min after intubation. Blood pressure and heart rate both showed a significant fall from baseline values 10 min after intubation in both buprenorphine groups, with the changes being greater in the 5 micrograms.kg-1 group. We recommend the use of 2.5 micrograms.kg-1 buprenorphine for attenuation of the hypertensive response to intubation in major abdominal surgery. PMID- 8712330 TI - Attitudes to pain and pain relief in adult surgical patients. AB - Pain relief after surgery is frequently inadequate. In the last few years much research has been devoted to improving the situation. Unfortunately, very little work has been undertaken to explore the patients' contribution to pain management. The beliefs and attitudes held by patients when they enter the hospital environment may be responsible in some instances for their not achieving optimal pain relief from the available techniques. We have studied some of these attitudes with a survey of 180 adult patients admitted for elective surgery. We found that most patients still expect pain following surgery. However, they are not afraid to ask for analgesics when in pain and do not attribute pain to their own wrong doing. There are, however, some patients who appear to have 'deviant' pain beliefs that could hinder their appropriate use of analgesics. Sadly, it is impossible to identify these patients according to age, gender, socio-economic group or previous experience of pain or surgery. PMID- 8712331 TI - Molecular sieves: an alternative method of carbon dioxide removal. PMID- 8712332 TI - Hereditary angioedema: prophylaxis management in the puerperium. PMID- 8712333 TI - The electronic anesthetic logbook: the value of supervision. PMID- 8712334 TI - Sternomental distance--a useful predictor of difficult intubation in patients with cervical spine disease? PMID- 8712335 TI - Laryngeal mask insertion, cricoid pressure and manual in-line stabilisation. PMID- 8712336 TI - Reinforced laryngeal mask airway and nasal surgery. PMID- 8712337 TI - Another foreign body in a laryngeal mask airway. PMID- 8712338 TI - Hot air warming devices--a potential hazard. PMID- 8712339 TI - Kinking of tracheal tubes. PMID- 8712340 TI - Accuracy of blind placement of percutaneous tracheostomy. PMID- 8712341 TI - Obstruction of tracheostomy tube by tracheal wall after percutaneous tracheostomy. PMID- 8712342 TI - Transtracheal ventilation. PMID- 8712343 TI - Capnometry error. PMID- 8712344 TI - Drugs in status epilepticus. PMID- 8712345 TI - Attitudes and preferences of patients to routes of analgesic administration. PMID- 8712346 TI - Depth of the epidural space in children. PMID- 8712347 TI - ARDS or pulmonary oedema? PMID- 8712348 TI - Potentially fatal subclavian artery haemorrhage. PMID- 8712349 TI - Anaesthesia for quinsy tonsillectomy. PMID- 8712350 TI - Clonidine--and adjunct in isoflurane N2O/O2 relaxant anaesthesia. PMID- 8712351 TI - Valve perforation associated with pulmonary catheter insertion. PMID- 8712352 TI - A close shave: air embolism following laceration of central venous catheter. PMID- 8712353 TI - Injury to an anaesthetist during patient transfer. PMID- 8712354 TI - Silver(I)-mediated separations by capillary zone electrophoresis and micellar electrokinetic chromatography: argentation electrophoresis. AB - The addition of Ag(I) to the run buffer in capillary zone electrophoresis (CZE) and micellar electrokinetic chromatography (MEKC), containing sodium dodecyl sulfate, applies the principles of argentation chromatography to electrophoretic separations and is termed "argentation electrophoresis". This technique is shown to provide a complementary method to CZE and MEKC for the separation of specific types of solutes that selectively complex with Ag(I). Baseline resolution in the CZE separation of nine sulfonamides is achieved by the addition of 50 mM silver nitrate to the run buffer. Retention mechanisms in MEKC separations can also be manipulated by the addition of Ag(I) to the micellar solution. Only slight resolution of a pair of sulfonamides was achieved under normal MEKC conditions. Upon the addition of Ag(I) to the mobile phase containing SDS micelles, baseline resolution of the compounds is shown. The retention order and resolution of five sulfonamides changed significantly when 25 mM Ag(I) was added to the SDS containing buffer. The use of Ag(I) addition in MEKC is also applied to the separation of various other compounds that show selectivity toward Ag(I) complexation, including S-containing heterocycles and vitamin D compounds. The effect of the addition of Ag(I) on the elution range in MEKC is also investigated. A steady increase in the elution range is seen upon increasing the concentration of Ag(I). With a constant percentage of organic modifier (15%), the addition of higher concentrations of silver nitrate (25-50 mM) results in elution ranges greater than 12. The results using Ag(I) as buffer additives in MEKC are also compared to studies utilizing a mixed counterion surfactant of sodium/silver dodecyl sulfate. PMID- 8712355 TI - Automated analytical system for the examination of protein primary structure. AB - This paper describes an automated analytical system for the examination of protein primary structure in which (i) the target protein is first purified by immunoaffinity chromatography, (ii) subsequent chromatographic and chemical reaction steps in the sequencing process are directly coupled, (iii) buffer exchange between these unit operations is achieved while the protein is absorbed on a mixed bed of strong ion exchange sorbents, (iv) proteolysis occurs in an immobilized trypsin column having a 10-1000 fold-excess of enzyme, (v) the tryptic digest is directly transferred to a perfusion dilute capture column where it is concentrated and rapidly desalted, and (vi) peptides eluted from the dilute capture column and analytical microbore and capillary perfusion reversed-phase chromatography columns are analyzed by either single-stage mass spectrometry (MS) or tandem MS/MS. Protein structure variants were easily recognized, and in the case of hemoglobin (Hb) S, the site of variation from Hb A0 was verified. PMID- 8712356 TI - Pulsed gas introduction for increasing peptide CID efficiency n a MALDI/quadrupole ion trap mass spectrometer. AB - A pulsed valve was used for studying the effects of introducing heavy gases at different stages of operation of a quadrupole ion trap and for increasing the efficiency of collision-induced dissociation (CID) of peptide ions at low values of the Mathieu parametere qz. When amounts of heavy gases comparable to that of the helium buffer gas were introduced during the ion trapping, ion isolation, and mass spectral recording stages, the effects on performance were generally small or negative. However, injection of heavy gases during CID provided considerable improvement in fragmentation efficiency that depended upon the particular gas used, its mass and pressure, and the amplitude of the excitation voltage. Efficient peptide fragmentation could be demonstrated for values of qz as low as 0.05, which permitted trapping of low-mass product ions and (in many cases) full recovery of the amino acid sequence. In this report, examples are provided of monoisotopic tandem mass spectra of peptide ions with masses up to 1570 Da. PMID- 8712357 TI - Selection of reference or test materials for the validation of atomic absorption food analysis methods. AB - A strategy to select reference or other test materials on which method validation experiments should be performed is developed for the AAs analysis of foodstuffs. Representative materials are selected for each category of food (meat, cereals, vegetables, etc.) and for the complete food spectrum. A principal component analysis and two cluster analysis procedures are applied to accomplish this. PMID- 8712358 TI - A self-assembled monolayer for the binding and study of histidine-tagged proteins by surface plasmon resonance. AB - This paper reports the generation of a self-assembled monolayer (SAM) that selectively binds proteins whose primary sequence terminates with a His-tag: a stretch of six histidines commonly incorporated in recombinant proteins to simplify purification. The SAM was prepared by the adsorption onto a gold surface of a mixture of two alkanethiols: one thiol that terminated with a nitrilotriacetic acid (NTA) group, a group that forms a tetravalent chelate with Ni(II), and a second thiol that terminated with a tri(ethylene glycol) group, a group that resists protein adsorption. His-tagged proteins bound to the SAM by interaction of the histidines with the two vacant sites on Ni(II) ions chelated to the surface NTA groups. Studies with model proteins showed the binding was specific for His-tagged proteins and required the presence of Ni(II) on the surface. Immobilized His-tagged proteins were kinetically stable in buffered saline at pH 7.2 but could be desorbed by treatment with 200 mM imidazole. Surface plasmon resonance studies for two model systems showed that His-tagged proteins adsorbed on the NTA-SAM retained a greater ability to participate in binding interactions with proteins in solution than protein immobilized in a thin dextran gel layer by covalent coupling. PMID- 8712359 TI - Laser-excited atomic fluorescence spectrometry in a pressure-controlled electrothermal atomizer. AB - A theoretical model was developed to describe the loss of analyte atoms in graphite furnaces during atomization. The model was based on two functions, one that described the supply of analyte by vaporization, and another that described the removal of the analyte by diffusion. Variation in working pressure was shown to affect the competition between these two processes. Optimal atomization efficiency was predicted to occur at a pressure where the supply of the analyte was maximized, and gas phase interactions between the analyte and matrix were minimized. Experiments to test the model included the direct determination of phosphorus and tellurium in nickel alloys and of cobalt in glass. In all cases, reduction in working pressure from atmospheric pressure to 7 Pa decreased sensitivity by 2 orders of magnitude, but improved temporal peak shape. For the atomization of tellurium directly from a solid nickel alloy, and the atomization of cobalt from an aqueous solution, no change in sensitivity was observed as the working pressure was reduced from atmospheric pressure to approximately 70 kPa. If a reduction in working pressure affected only the diffusion of the analyte, poorer sensitivity should have been obtained. Only a commensurate increase in analyte vaporization could account for maintained sensitivity at lower working pressures. Overall, analyte vaporization was not dramatically improved at reduced working pressures, and maximum atomization efficiency was found to occur near atmospheric pressure. PMID- 8712360 TI - Correlation of kinetic energy losses in high-energy collision-induced dissociation with observed peptide product ions. AB - In collision-induced dissociation, some of an incident parent ion's kinetic energy is converted into internal energy upon collision with a neutral target. The kinetic energy lost is related to the amount of internal energy deposited into any individual ion. To see dissociations of different critical energies on the same time scale, different amounts of internal energy need to be deposited. This should be reflected in the kinetic energy lost by the parent ion in the formation of different product ions. Variable amounts of energy loss in the formation of different peptide product ions are reported here. It is seen that different product ion types (b, y, a) show ordered patterns of energy losses. A greater energy loss is observed for the formation of b-type product ions than for y-type, and even greater energy losses are observed for the formation of a-type product ions. A very good correlation between ion type energy loss and ion mass is observed. Thus, measuring the energy losses in the formation of product ions may provide a means for classifying the product ion type. PMID- 8712361 TI - Parent ion scans of unseparated peptide mixtures. AB - The nanoelectrospray ion source (NanoES) developed recently has proven to be an excellent tool for the sequencing of peptides out of unseparated mixtures. At lower levels of analyte, however, it is increasingly difficult to distinguish the peptide ions in the spectrum, limiting the overall sensitivity of the procedure. Scans for the parents of common fragmentation products of peptides such as immonium ions allow determination of peptide ion masses even when these ions have signal-to-noise ratios of < 1 in the mass spectrum. With this technique, concentration limits for sequencing peptides could be improved to < 5 fmol/microL in favorable cases. Parent ions scan were used to identify posttranslational modifications of peptides in mixtures, with phosphorylation and glycosylation as examples. Detecting phosphorylation by parent scans increased sensitivity by 1-2 orders of magnitude. Modified peptides can be subjected to tandem mass spectrometry in the same experiment to localize the modification. The present approach expands the range of application of nanoelectrospray and complements established liquid chromatography/mass spectrometry methods. PMID- 8712362 TI - "Colored" noise waveforms and quadrupole excitation for the dynamic range expansion of Fourier transform ion cyclotron resonance mass spectrometry. AB - Fourier transform ion cyclotron resonance (FTICR) mass spectrometry offers unparalleled analytical performance in most regards but has a dynamic range of typically no better than 10(2)-10(3). This limitation reportedly arises from two opposing constraints, involving the maximum number of ions that can be effectively trapped (10(6)-10(7)) and the minimum number of ions required to produce a detectable signal (10(2)-10(3)). A potential solution to this dynamic range limitation is presented, based on the application of selected-ion accumulation using quadrupole excitation. We show that lower concentration species can be effectively accumulated in the FTICR trapped ion cell, while the more abundant species are continually removed by the application of quadrupolar excitation in the form of band-limited or "colored" noise waveforms. The result is that "room" is made in the cell for lower abundance species, even during extended accumulation periods. This approach was demonstrated with mixtures of the bovine proteins, insulin, ubiquitin, and cytochrome c. For normal accumulation, the dynamic range was approximately 100. The application of selected-ion accumulation in the form of colored noise allowed the extension by 2 orders of magnitude and the detection of species of 1 x 10(-8) M concentration from a solution also containing another component at 9 x 10(-5) M. With this method, a putative new low abundance variant of bovine insulin was observed, and selected-ion accumulation and subsequent collisionally activated dissociation were used for its identification. Dipolar magnetron excitation was also explored to enhance selected-ion accumulation and was found to reduce the amount of buffer gas required for complete removal of the undesired species by a factor of 5. Further possible improvements are discussed, as are the complications due to the required balance between magnetron and cyclotron damping rates. PMID- 8712363 TI - Isotopic assignment in large-molecule mass spectra by fragmentation of a selected isotopic peak. AB - For large ( > 5 kDa) ionic species, Fourier transform ion cyclotron resonance instruments yield by far the highest mass accuracy. However, this can be compromised by misassignment of the isotopic content based on predicted natural abundances of the isotopic peaks. As an alternative method independent of natural abundance variations, high-resolution isolation and dissociation of a single isotopic peak yields a distribution of isotopic peak abundances characteristic of the isotopic content of the precursor peak. Accuracy is enhanced if the precursor peak is abundant and of minimum heavy isotope content, and if the product species is abundant and of intermediate mass. In addition, such spectra of the highest mass products are useful for identifying complementary product pairs, a key step in sequencing proteins and nucleotides. PMID- 8712364 TI - Simplifying the exoglycosidase digestion/MALDI-MS procedures for sequencing N linked carbohydrate side chains. AB - Exoglycosidase digestion coupled with matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) is an effective technique for sequencing the N linked carbohydrate side chains of a glycoprotein. However, the buffers currently used in the enzymatic procedures are detrimental to MALDI-MS, and thus desalting is required before the digestion products can be analyzed. We demonstrate that a 25 mM ammonium acetate solution adjusted to the proper pH can replace the normal exoglycosidase digestion buffers. The use of these ammonium acetate solutions permits direct MALDI-MS analysis of the digestion mixture without desalting. More importantly, we show that many of the commonly used exoglycosidases retain both their activity and their specificity under these conditions. PMID- 8712365 TI - Effect of impurities on the matrix-assisted laser desorption mass spectra of single-stranded oligodeoxynucleotides. AB - The effect of impurities on the analysis of single-stranded DNA oligomers by the technique of matrix-assisted laser desorption/ionization with time-of-flight mass spectrometry has been studied using the matrix 3-hydroxypicolinic acid and 355-nm pulsed light. By mixing the DNA oligomers with different concentrations of impurities and recording mass spectra, limits are set on the tolerable level of a given impurity in a sample. The tolerance limits for sodium chloride, potassium chloride, sodium acetate, sodium fluoride, sodium dodecyl sulfate (SDS), and manganese(II) chloride were found to be approximately 10(-2) M. It was found that magnesium salts degraded the mass spectrum at much lower levels of 10(-4) M. The organic compounds tris(hydroxymethyl)aminomethane (Tris), urea, dithiothreitol (DTT), glycerol, and ethylenediaminetetraacetic acid (EDTA), when present as its ammonium salt, were tolerable at concentrations into the range of 0.25-0.5 M, while the organic polyamine compound spermine substantially degraded the mass spectrum at concentrations above 10(-2) M. When comparing these results for DNA analysis with previously reported limits for protein analysis, large differences are seen for some of the impurities tested. PMID- 8712366 TI - Professor Tomas Pexieder (6.6.1941-28.10.1995). PMID- 8712367 TI - Postnatal development of ovine seminiferous tubules: an electron microscopical and morphometric study. AB - Corresponding to the increasing testicular volume and the histological appearance of the testicular parenchyma, the postnatal ontogenesis of the ovine testis can be divided into five phases. During the prebubertal period (phases 1-III), seminiferous tubules are solid and contain supporting (pre-Sertoli) cells as well as up to three types of germ cells: prespermatogonia I, II and spermatogonia precursor cells. In phase I, only prespermatogonia I are present and can usually be observed at the center of the seminiferous tubules. During phase II, prespermatogonia I migrate towards the basal lamina, divide and become prespermatogonia II. Those prespermatogonia I which are not successful in establishing contact with the tubular basal lamina degenerate. In phase III, prespermatogonia II divide and differentiate into cells which function as stem cells for spermatogenesis. Morphometric data corroborate the assumption of two types of prespermatogonia in the postnatal prepubertal ovine testis. Prespermatogonia I have nuclear volumes of about 480 microns 3 and cellular volumes of about 1200 microns3. In prespermatogonia II both volumes increase to about 920 microns3 and 1800 microns3 respectively. Adult A-spermatogonia are significantly smaller and possess an average nuclear volume of about 340 microns3 and an average cellular volume of about 800 microns3. Concomitanty with the formation of the tubular lumen in puberty (phase IV), supporting cells differentiate morphologically into typical Sertoli cells. Developmental events in the germ cell population are not yet synchronized. Adulthood (phase V) is characterized by complete spermatogenesis with all stages of the seminiferous epithelial cycle. PMID- 8712368 TI - Vascular architecture of the human fetal adrenal gland: a SEM study of corrosion casts. AB - The vascular architecture of adrenal glands was investigated in human fetuses aged from 16 to 24 weeks, using microcorrosion casts and scanning electron microscopy. The fetal adrenals showed an arrangement of blood vessels remarkably similar to that described in adult glands. There was a clear centripetal pattern from superficial arteries and their branches, via irregular capillaries of the subcapsular plexus and definitive cortex, and then via the radial sinusoids and venous sinuses of the fetal cortex, to the central vein. Rare medullary arterioles traversed the cortex to break up into small local capillary networks in the central region of the gland. Some superficial capillaries were drained by occasional subcapsular veins. No portal system was observed in the fetal adrenals. PMID- 8712369 TI - Histomorphological and chemico-physical analyses of the mineral matrix of micropetrotic human bone. AB - Micropetrotic areas of human bone were analyzed with reference to their distance from blood vessels and to the age of the subjects. Undecalcified bone sections were treated with various solvent, soaking and etching substances to establish the nature of the material occluding the osteocytic canalicular cavities, and were examined by means of microradiographic and microdurimetric methods and X-ray microanalysis to evaluate the degree of mineralization in the bone matrix. Since it was only after strong etching with HCl that the canalicular network became visible under light and scanning electron microscopy, it is clear that the debris filling the canalicular network consists almost entirely of mineral substance. The degree of mineralization of micropetrotic bone is high because it is always a more mature type of bone, but the mineral content of the matrix and the Ca/P ratio do not differ significantly from those of neighbouring bone where the canalicular network is fully pervious. PMID- 8712370 TI - Relationship between the proliferation of keratinocytes cultured in vitro and prostaglandin E2. AB - In the growth of keratinocytes "in vitro", PGE2 seems to play an important role. We have shown that in fibroblast-keratinocyte co-cultures, indomethacin, employed at concentrations which inhibit the PGE2 synthesis, reduced the proliferation of epidermal cells. This effect was reversed by an exogenous PGE2 addition to the culture media. To better understand the relationship between keratinocytes and the autacoid, we have tested PGE2 at various concentrations in different cultural conditions, that is, epidermal cells were grown on a 3T3-J2 feeder layer, without fibroblasts and with a 3T3-J2 conditioned medium. We observed an increase in keratinocyte proliferation induced by the autacoid alone in the presence of fibroblasts, while a severe inhibitory effect was relieved when dermal cells or the conditioning medium were absent. The lack of fibroblasts and their products in the culture medium modified the morphology of keratinocytes cultured in vitro. PGE2 induced significant morphological and morphometrical variations only if added to the conditioning medium. The autacoid decreased the expression of 66 kDa protein, if cells were grown in the presence of fibroblasts or with conditioning medium, whereas it completely inhibited this keratin and those of 60, 54 kDa if cells were cultured only with a basal medium. From morphometrical and electrophoretical data we can suppose that PGE2 inhibits cell differentiation. Thus PGE2 action on keratinocytes seems to be strictly related to the presence of dermal cells. PMID- 8712371 TI - [Adhesions and interference images of unfixed erythrocytes]. AB - For the analysis of deformability in microcirculatory investigations an exact understanding of red blood cell (RBC) geometry is required. To extend knowledge we introduce a new morphological feature of resting unfixed erythrocytes by means of an inverted reflection contrast microscope (RCM). By assessing the interference patterns caused by RCM erythrocytes can be classified according to the depth of their central concavity which depends on the flexibility of the RBC. Moreover, the RBC adhesion can be directly observed. We found out that: 1. Five types of normocytes can be distinguished in RCM. 2. In phase contrast the size distribution of RBC without central concavity (type 5, 11.53% of all normocytes) shows peaks at 48 microns2 and 52 microns2. 3. Image analysis reveals two size categories of relative adhesion areas. One category consists of type 1 and 2 (relative adhesion area 25.63%), the other of the types 3 to 5 (relative adhesion area 39.91%). Besides, RCM allows the reliable identification of pathologic erythrocytes in unstained specimens. PMID- 8712372 TI - Histological and ultrastructural study of the lingual epithelium of the juvenile Pacific ridley turtle, Lepidochelys olivacea (Chelonia, Cheloniidae). AB - Histological and ultrastructural studies ot the dorsal lingual epithelium of the juvenile Pacific ridley turtle, Lepidochelys olivacea, were performed by light and electron microscopy, and the results were compared to those of freshwater turtles in order to clarify the relationship between the histological and cellular differences of the lingual epithelium and the habitat of the turtles. The tongue of the juvenile Pacific ridley turtle is triangular with a round apex when viewed from above, but it appears flattened in lateral view. Scanning electron microscopy (SEM) revealed no lingual papillae on the dorsal surface of the tongue. Instead, transverse plicae are found on the surface of the body and the radix. The surface of the apex is smooth. Microridge-like structures are present on the surfaces of the cells, and the cell margins are thickened. The mucosal epithelium is keratinized, stratified squamous with a relatively thick layer of desquamating cells. Cells of the basal and deep intermediate layers appear elliptical in shape; and their nuclei are elliptical and centrally located. Numerous desmosomes join the processes of adjacent cells; and hemidesmosomes anchor the basal cells to the basal lamina. The cytoplasm of these cells contains mitochondria, free ribosomes, rough endoplasmic reticulum, vacuoles, and bundles of tonofilaments. Cells and their nuclei in the intermediate layer display gradual flattening. In the shallow intermediate layer, the cells are significantly flattened, with nuclei condensed or absent. The cytoplasm contains many tonofibrils or bundles of tonofilaments, free ribosomes and keratohyalin granules, with numerous ribosomes attached to their surfaces. A few collapsed mitochondria are visible. Cell membranes of the shallow intermediate cells are smooth and attached to those of adjacent cells by desmosomes. The keratinized layer is located on top of the shallow intermediate layer, and consists of significantly flattened cells lacking nuclei and filled with keratin fibers. Very fine cellular processes joined by desmosomes are visible. The desquamating cells located on top of the keratinized layer contain keratin fibers that are somewhat thicker than tonofibrils and tonofilaments, and clearly distinguishable individually. The microridge-like structures visible by SEM could be attributed to the persistence of cells formed in underlying layer. In conclusion, the histology of the lingual epithelium of the juvenile Pacific ridley turtle differs significantly from that of the adult freshwater turtle in spite of the similarity of the gross morphology of their tongues. PMID- 8712374 TI - Scanning electron microscopy of human cerebral meningeal stomata. AB - The human dura mater and pia mater were studied by using a scanning electron microscope and a computer image processing system (C.I.P.). The human cerebral meningeal stomata are located between the mesothelial cells of the cerebral meninges. They are round or oval in shape with diameters of 0.33-2.98 microns. The cerebral meningeal stomata are stable structures, scattered or clustered together. Their density in the dura mater is greater than in the pia mater (P < 0.01), and they are regularly distributed. The statistical analysis showed that the stomata diameter and distribution density in the dura mater are 1.34 microns and 381.55/0.1 mm2; while in the pia mater they are 0.88 micron and 195.06/0.1 mm2 respectively. The cerebral meningeal stomata are probably part of the cerebral prelymphatic capillary system, which undertakes the cerebral lymph drainage because there are no lymphatic vessels in the brain although yet there is lymph drainage. Thus, we believe that the cerebral meningeal stomata are involved in maintaining the physiological function of the brain as part of the cerebrospinal fluid (CSF) which absorbs the cerebral interstitial fluid. PMID- 8712373 TI - Presence and structure of anal and vaginal tonsils in the laboratory shrew (Suncus murinus). AB - In all the laboratory shrews (Suncus murinus) examined, a pair of tonsil-like structures was found at the boundary between the anus and the ostium urogenitoanalis. Moreover, the same tissues were recognized between the vagina and the ostium urogenitoanalis in 9 of the 13 females examined. They indicated clear epitheliolymphocyte symbiosis around both mucosal crypts (anal and vaginal crypts) and outer lymph nodules including the germinal centers, and were demarcated from other tissues by the fibrous capsule. Two kinds of tonsil-like structures in this study were found to be homologous with some of the tonsils in the fauces of other mammals. These will be therefore referred to as the anal and vaginal tonsils. PMID- 8712375 TI - Accessory ossifications in the ulno-carpal wrist region. AB - Accessory ossifications in the ulno-carpal transition such as ossa styloidea or triangularia represent a rare finding, especially among younger individuals. The present results of radiological and morphological investigations show that up to 50% of the ossa are located palmar to the styloid process. Most of them have a sharply bordered contour and are less than 10 mm in length. An aetiological classification of accessory ossifications in the distal epiphyseal region is only possible when an obvious case history is available. Accessory skeletal elements are present among elderly individuals as a result of sclerotic alterations of the ulno-carpal joint elements. Among younger individuals, however, they point to a pathological aetiology and can therefore assume clinical and diagnostic relevance in cases involving inflammatory rheumatic and traumatic alterations accompanied by vague discomfort in the proximal wrist region. PMID- 8712376 TI - Retroesophageal right subclavian artery originating from the aortic arch distal and dorsal to the left subclavian artery. AB - In the cadaver of a Japanese 79 year-old man a retroesophageal right subclavian artery was observed to be derived from the arch of the aorta slightly distal and dorsal to the left subclavian artery. Its origin formed Kommerell's arterial diverticulum (50 mm in circumference), and it passed between the esophagus and the vertebral column and continued to the right to become the axillary artery. No right recurrent laryngeal nerve was observed. There was a right ansa subclavia around the subclavian artery. Although this anomaly is relatively rare, it is important as a cause of dysphagia lusoria. PMID- 8712378 TI - The superficial ulnar artery originating from the axillary artery. AB - An anomalous superficial ulnar artery was found during anatomical dissection in the right arm of an 83-year-old Japanese woman. It originated in the axillary artery, crossed over the median nerve, coursed ventral to the median nerve and the brachial artery, but superficial to the bicipital aponeurosis and the flexor muscles. At the palm it formed the superficial and deep palmar arches together with the branches of the radial artery. The brachial artery divided into the radial and common interosseous arteries in the cubital fossa. PMID- 8712377 TI - Partial agenesis of the azygos vein: a case report. AB - In the cadaver of an 86 year old man the inferior segment of the azygos vein could not be found. Furthermore, a normally developed hemiazygos vein drained the right and left intercostal veins from T 10 to T 6. This vessel finally curved towards the right to reach the superior vena cava after having drained the right superior intercostal veins. The left superior intercostal vein ended in a short vessel draining into the left brachiocephalic vein. This condition may be represented in a standard chest radiograph by the so-called "aortic nipple". Agenesis of the azygos vein, suspected because of the presence of this radiological sign, should be confirmed in the living by means of computerized tomography. This can clarify the anatomy of the mediastinal vessels precisely. Embryological pattern of the azygos system accounting for its possible defects is discussed. PMID- 8712379 TI - A case of duplicated right renal artery and triplicated left renal artery. AB - A rare anomaly of the vessels supplying the kidney of a 65 year-old male cadaver was found during a routine dissection. The right kidney received two renal arteries from the aorta that were similar in diameter, both entering through the hilus. The left kidney had three arteries originating from the aorta, one at its usual hilar position and two entering the renal cortex at its upper and lower poles. The upper pole of the left kidney also gave rise to an additional tributary of the renal vein. PMID- 8712380 TI - Arterial vascularization of the abdominal and pelvic regions in the degu, Octodon degus (Rodentia, Octodontidae). AB - The abdominal and the pelvic arterial supply of the degu, Octodon degus, were described in 19 specimens (10 males and 9 females) from a laboratory strain. Animals were injected through the left ventricle of the heart with coloured latex solution and subsequently dissected under a surgical microscope. The arrangement and variations of the visceral and muscular arteries were described. Comparative analysis among rodents revealed that, even though there was a wide range of interspecific variation in the characters considered, there were several constant anatomical features with phylogenetic value. In this sense, results on the degu and the guinea pig, Cavia porcellus, showed that these hystricognaths share particular features that differentiate them from the sciurognaths analysed to date. The distinguishing characteristics between these two groups of rodents were found in the arrangement of the coeliacomesenteric trunk, and in the cranial abdominal, right gastric, caudal mesenteric, and deep circumflex iliac arteries. The phylogenetic meaning of particular angiological features in rodents were commented. PMID- 8712381 TI - Medical ethics and legal standards. PMID- 8712382 TI - A pilot study of the effects of a perflubron emulsion, AF 0104, on mixed venous oxygen tension in anesthetized surgical patients. AB - A pilot study of a perfluorochemical (PFC) emulsion was undertaken to determine whether administration of a perflubron emulsion could result in measurable changes in mixed venous oxygen tension. Seven adult surgical patients received a 0.9-g PFC/kg intravenous dose of perflubron emulsion after acute normovolemic hemodilution (ANH). Hemodynamic and oxygen transport data were collected before and after ANH, immediately after PFC infusion, and at approximate 15-min intervals throughout the surgical period. There were no clinically significant hemodynamic changes associated with the administration of the PFC emulsion. There was a significant increase in mixed venous oxygen tension (PVO2) after the PFC infusion, while cardiac output and oxygen consumption were unchanged. As surgery progressed, the hemoglobin concentration decreased with ongoing blood loss while PVO2 values remained at or above predosing levels. Peak perflubron blood levels were 0.8 g/dL immediately postinfusion, and approximately 0.3 g/dL at 1 h. This pilot study demonstrates that administration of perflubron emulsion results in measurable changes in mixed venous oxygen tension during intraoperative ANH. PMID- 8712383 TI - The effect of ulinastatin, a human protease inhibitor, on the transfusion-induced increase of plasma polymorphonuclear granulocyte elastase. AB - Ulinastatin (UTI), a human protease inhibitor, inhibits polymorphonuclear granulocyte elastase (PMNE) release from granulocytes stimulated by several pathologic inflammatory processes. Blood transfusions increase plasma PMNE concentrations. We evaluated the effects of UTI on the transfusion-induced increase in PMNE. Thirty-nine patients undergoing gastrectomy were divided into four groups: Group A (n = 8), blood transfusion (-), UTI (-); Group B (n = 9), blood transfusion (+), UTI(-); Group C (n = 12), blood transfusion (-), UTI(+); and Group D (n = 10), blood transfusion (+), UTI(+). UTI of 300,000 U was infused in Groups C and D. White blood cell count (WBC), segmented cell count, plasma PMNE concentration, and PMNE release (PMNE concentration/segmented cell) were measured before, at the end of, and at 1 and 3 days after the operation. Although no significant differences were seen in WBC and segmented cell count among the groups, plasma PMNE concentration and PMNE release in Group B (transfusion without UTI) were significantly more than in Group A (no treatment) and Group D (transfusion with UTI) at the end of the operation, but the concentrations of the variables were not significantly different between Groups A and C (only UTI treatment). These results indicate that a single injection of 300,000 U of UTI inhibited transfusion-induced PMNE increase at the end of the operation. Thus, the present results may facilitate future studies testing the effects of UTI given at larger doses, or by a continuous administration, on the prevention of organ derangement after blood transfusion. PMID- 8712384 TI - Perioperative changes in cardiac autonomic control in patients receiving either general or local anesthesia for ophthalmic surgery. AB - To investigate whether local (LA) or general anesthesia (GA) provides more perioperative stability in cardiac autonomic tone, we analyzed heart rate variability (HRV) by means of spectral analysis, intraoperatively and up to 3 h postoperatively, in otherwise healthy patients scheduled for cataract surgery. Fourteen patients received GA (premedication: clorazepate; anesthetic induction: propofol, alfentanil, atracurium; anesthetic maintainance: isoflurane, alfentanil; airway management: laryngeal mask airway), and 14 patients received LA (retrobulbar block with bupivacaine/mepivacaine). In the GA group, total power, as an indicator of cardiac autonomic activity, was significantly reduced intraoperatively and increased slowly during the postoperative period. The ratio of low frequency/high frequency (LF/HF) power, indicating cardiac sympathovagal balance, did not change significantly intraoperatively and remained stable in the postoperative period. The LA group showed no significant changes in total power intraoperatively. Postoperatively, heart rate and the LF/HF power ratio were significantly increased in the LA group compared to the GA group. GA as described above was associated with intra- or postoperative stability of the cardiac sympathovagal balance. Thus GA had no disadvantageous effects on the perioperative cardiac autonomic tone during ophthalmic surgical procedures compared to LA in otherwise healthy patients. PMID- 8712385 TI - Detection of gas embolism by transesophageal echocardiography during laparoscopic cholecystectomy. AB - Using transesophageal echocardiography (TEE), 16 patients (ASA physical status I III), undergoing laparoscopic cholecystectomy, were assessed for the occurrence of episodes of gas embolism and cardiovascular changes related to those emboli. The long-axis four-chamber view was monitored continuously, except for predetermined intervals where the transgastric short-axis view was obtained to derive the end-diastolic area (EDA), the end-systolic area (ESA), and the ejection fraction (EF). In one patient, we monitored the longitudinal view of the superior and the inferior vena cava. The monitoring of the patients also included: heart rate (HR), mean arterial pressure (MAP), arterial saturation by pulse oximetry (Spo2), end-tidal CO2 (ETCO2), minute ventilation (VE), and peak inspiratory pressure (PIP). Embolic events were defined as the appearance of gas bubbles in the right cardiac chambers. We observed gas embolism in 11/16 patients (five during peritoneal insufflation and six during gallbladder dissection). Using the longitudinal view of the superior and inferior vena cava (IVC), we found that these emboli were transmitted through the IVC. No episode of cardiorespiratory instability (decrease in MAP > or = 10 mm Hg, Spo2 < 90%) was observed. There was no significant difference in cardiorespiratory variables between patients who presented gas embolism (n = 11) and patients who did not (n = 5) during the studied period. In this small group of patients, we conclude that gas embolism occurs commonly during laparoscopic cholecystectomy but that these gas emboli cause minimal cardiorespiratory instability. PMID- 8712386 TI - Head rotation during internal jugular vein cannulation and the risk of carotid artery puncture. AB - We undertook a prospective laboratory study to examine the effect of head position on the relative positions of the carotid artery and the internal jugular vein (IJV). Volunteers (n = 12) from departmental staff, 18-60 yr of age, who had never undergone cannulation of the IJV underwent imaging of their IJV and carotid artery. With the subject in a 15 degrees Trendelenburg position, two-dimensional ultrasound images of the IJV and the carotid artery were obtained on the left and right sides of the neck at 2 and 4 cm from the clavicle along the lateral border of the sternal head of the sternocleidomastoid muscle at 0 degrees, 40 degrees, and 80 degrees of head rotation from the midline. The percent overlap of the carotid artery and IJV increased significantly at 40 degrees and 80 degrees head rotation to both the right and left (P < 0.05). Data from 2 and 4 cm above the clavicle did not differ and were pooled. The percent overlap was larger on the left than the right only with 80 degrees of head rotation (P < 0.05). The increased overlap of carotid artery and IJV with head rotation > 40 degrees increases the risk of inadvertent puncture of the carotid artery associated with the common occurrence of transfixion of the IJV before it is identified during needle withdrawal. The IJV frequently collapses with needle insertion. This may result in puncture of the posterior wall of the vessel, and thus of the carotid artery when the two vessels overlap. To decrease this risk, the head should be kept in as neutral a position as possible, that is < 40 degrees rotation, during IJV cannulation. PMID- 8712387 TI - Survey of laryngeal mask airway usage in 11,910 patients: safety and efficacy for conventional and nonconventional usage. AB - A survey of laryngeal mask airway (LMA) usage was conducted to provide general information about safety and efficacy with special emphasis on controversial issues such as positive pressure ventilation (PPV), prolonged anesthesia, and laparoscopic and nonlaparoscopic intraabdominal surgery. During the 2-yr study period, of the 39,824 patients who underwent general anesthesia, 11,910 (29.9%) patient airways were managed with the LMA. Forty-four percent underwent PPV. Placement was successful in 99.81%, and in 23 patients the LMA was abandoned in favor of the tracheal tube (TT). Use of the LMA for any intraabdominal procedure was considered nonconventional and occurred in 2222 (18.7%) patients. On 579 occasions procedures lasted > 2 h. A total of 44 critical incidents were documented. Eighteen (0.15%) were related to the airway and none required intensive care management. There were 26 critical incidents not related to the airway which resulted in two admissions to the intensive care unit and one death. There were three cases of failed tracheal intubation managed with the LMA. This survey demonstrates that the LMA technique is safe and effective for both spontaneous and controlled ventilation. Use of the LMA for gynecologic laparoscopy, gynecologic laparotomy, and procedures > 2 h also appears safe. PMID- 8712388 TI - Factors associated with excessive postoperative blood loss and hemostatic transfusion requirements: a multivariate analysis in cardiac surgical patients. AB - The purpose of this study was to prospectively evaluate whether heparin and protamine doses administered using a standardized protocol based on body weight and activated clotting time values are associated with either transfusion of hemostatic blood products (HBPs) or excessive postoperative bleeding. Analysis using 10 multiple logistic or linear regression models in 487 cardiac surgical patients included perioperative variables that may have an association with either transfusion of HBP and/or excessive postoperative chest tube drainage (CTD). Prolonged duration of cardiopulmonary bypass (CPB), lower pre-CPB heparin dose, lower core body temperature in the intensive care unit, combined procedures, older age, repeat procedures, a larger volume of salvaged red cells reinfused intraoperatively and abnormal laboratory coagulation results (prothrombin time, activated partial thromboplastin time, and platelet count) after CPB were associated with both transfusion of HBP and increased CTD. Female gender, lower total heparin dose, preoperative aspirin use and the number of HBPs administered intraoperatively were associated only with increased CTD, whereas a larger total protamine dose was associated only with perioperative transfusion of HBPs. Preoperative use of warfarin or heparin was not associated with excessive blood loss of perioperative transfusion of HBPs. In contrast to previous studies using bovine heparin, data from the present study do not support the use of reduced doses of porcine heparin during CPB. PMID- 8712389 TI - Neostigmine reversal of vecuronium neuromuscular block and the influence of renal failure. AB - The duration of clinical relaxation induced by vecuronium and reversal by neostigmine was studied in 40 patients with renal failure (RF) and 40 patients with normal renal function (NL) under general anesthesia. Patients were premedicated with flunitrazepam, and anesthesia commenced with fentanyl 1-2 micrograms/kg, thiopental 5-8 mg/kg, and vecuronium 0.1 mg/kg. Anesthesia was maintained with 60% nitrous oxide in oxygen, isoflurane 0.3%-1.0% end-tidal concentration, and 1 microgram/kg fentanyl every 20-30 min. Neuromuscular block was reversed by the administration of intravenous neostigmine 40 mg/kg at the time of reappearance of either two or four responses to the train-of-four (TOF) stimulation. Monitoring of neuromuscular function consisted of supramaximal TOF stimulation of the ulnar nerve and the evoked thumb response was registered using a force transducer. Spontaneous recovery time, reversal time, and the time to recovery of TOF ratio to 0.7 were recorded. RF did not prolong the vecuronium neuromuscular blocking effect, reversal was achieved at the same rate in NL as in RF, and the duration of reversal of neuromuscular blocking effect of vecuronium was not influenced by the time of administration of neostigmine. Therefore, the neuromuscular blocking effect of a tracheal intubating dose of vecuronium can be reversed at the same rate in patients with end-stage RF as in patients with normal kidney function. PMID- 8712390 TI - Influence and relative sensitivities of 50-Hz and 100-Hz tetanic stimuli on subsequent tetanic fade ratios in patients receiving vecuronium. AB - We studied the possible effects of repetitive (1-min interval) 50- and 100-Hz tetanic stimuli on 50-Hz and 100-Hz tetanic fade ratios (RF50HZ and RF100HZ). We also evaluated the sensitivity of the recorded responses to these two tests to assess residual neuromuscular block (isometric adductor pollicis mechanical activity), either during spontaneous recovery, or 15 min after neostigmine administration, in 22 adult anesthetized (thiopental, fentanyl, N2O/O2) patients receiving vecuronium. Two 50-Hz and two 100-Hz, 5-s duration, tetanic stimulations were randomly assessed at 1-min intervals: in a spontaneous (SPO) group (n = 11), when train-of-four (TOF) ratio spontaneously regained 0.7, and in a neostigmine (NEO) group (n = 11), 15 min after 40 micrograms/kg neostigmine was given intravenously at 25% return of control twitch tension. In the SPO group, when TOF ratio was 0.7, RF50HZ was 0.92 +/- 0.01 before and after subsequent tetanic stimulation, while RF100HZ was 0.48 +/- 0.05 and 0.47 +/- 0.05, respectively (not significant [NS]). In the NEO group, when TOF ratio was approximately 0.9, RF50HZ was 0.93 +/- 0.01 before and after subsequent tetanic stimulation, while RF100HZ was 0.80 +/- 0.02 and 0.78 +/- 0.02, respectively (NS). From patient to patient, both RF50HZ and RF100HZ were also identical. In conclusion, in patients receiving vecuronium, 1) 5-s, 50- and 100-Hz tetanic stimuli may be repeated without changes at 1-min intervals and, 2) in contrast to RF50HZ, recorded RF100HZ enables one to determine residual neuromuscular block during spontaneous recovery (P < 0.001) such as after neostigmine reversal (P < 0.05). PMID- 8712391 TI - The effect of esmolol pretreatment on the incidence of regional wall motion abnormalities during electroconvulsive therapy. AB - Electroconvulsive therapy (ECT) is associated with dramatic increases in arterial blood pressure and heart rate (HR) that may precipitate new left ventricular regional wall motion abnormalities (RWMAs) suggestive of myocardial ischemia. The purpose of this study was to investigate the effect of pretreatment with esmolol on the incidence of RWMAs after ECT. Thirteen patients served as their own controls and, in a random fashion, received a standard succinylcholine/methohexital anesthetic for one of two ECT sessions, and an identical anesthetic with esmolol 1 mg/kg for their other ECT session. Systolic (SBP), diastolic (DBP), mean arterial pressures (MAP) and HR were recorded after drug administration and before ECT and at 1-, 2-, 4-, 5-, 10-, and 15-min intervals after ECT. Echocardiograms were obtained at baseline, after drug administration, 1 min after ECT, and at recovery 15 min later. All patients had significant increases in SBP, DBP, and MAP at 1, 2 and 4 min after ECT versus baseline, whereas HR was significantly faster at all times in the control sessions. HR was significantly slower after anesthetic induction until 2 min after ECT in the esmolol versus the control group (P < 0.05). New RWMAs were seen in only 1 of 26 (4%) ECT sessions, despite the presence of baseline RWMAs in 31% of the patients. We conclude that contrary to previously reported data, new RWMAs after ECT are uncommon. Consequently, this study was unable to demonstrate any beneficial effect of pretreatment with esmolol on the incidence of ECT-induced RWMAs. PMID- 8712392 TI - Therapeutic suggestion has not effect on postoperative morphine requirements. AB - This study was designed to confirm the effect of therapeutic intraoperative auditory suggestion on recovery from anesthesia, to establish the effect of preoperative suggestion, and to assess implicit memory for intraoperative information using an indirect memory task. Sixty consenting unpremedicated patients scheduled for elective gynecologic surgery were randomly divided into three equal groups: Group 1 received a tape of therapeutic suggestions preoperatively, and the story of Robinson Crusoe intraoperatively; Group 2 heard the story of Peter Pan preoperatively and therapeutic suggestions intraoperatively; Group 3 heard the Crusoe story preoperatively and the Peter Pan story intraoperatively. A standardized anesthetic technique was used with fentanyl, propofol, isoflurane, and nitrous oxide. After surgery, all patients received patient-controlled analgesia (PCA) with a standardized regimen. In the 24 h postsurgery, morphine use was recorded every 6 h and at 24 h an indirect memory test (free association) was used to test for memory of the stories. Anxiety scores were measured before surgery and at 6 and 24 h postsurgery. There were no significant differences between groups for postoperative morphine use, pain or nausea scores, anxiety scores, or days spent in hospital after surgery. Seven of 20 patients who heard the Pan story intraoperative gave a positive association with the word "Hook," whereas 2 of 20 who did not hear the story gave such an association. Indirect memory for the Pan story was established using confidence interval (CI) analysis. (The 95% CI for difference in proportion did not include zero). No indirect memory for the Crusoe story could be demonstrated. This study did not confirm previous work which suggested that positive therapeutic auditory suggestions, played intraoperatively, reduced PCA morphine requirements. In contrast, a positive implicit memory effect was found for a story presented intraoperatively. PMID- 8712393 TI - The acute and residual effects of subanesthetic concentrations of isoflurane/nitrous oxide combinations on cognitive and psychomotor performance in healthy volunteers. AB - A blind, randomized, cross-over trial was conducted to determine the degree of psychomotor/cognitive impairment and the recovery profile produced by combinations of subanesthetic concentrations of isoflurane and nitrous oxide in healthy volunteers. In the experiment, subjects (n = 10) inhaled 100% oxygen placebo, 30% nitrous oxide in oxygen, and 0.2% and 0.4% isoflurane in oxygen, alone, and in combination with 30% nitrous oxide, in different sessions. Dependent measures included psychomotor and cognitive performance. Impairment was profound with the combination of inhaled anesthetics, and from an analysis of control conditions (the anesthetics alone), it appeared that isoflurane produced more impairment than did nitrous oxide. The time course of recovery was extremely rapid, with subjects returning to control-level functioning 5 min after cessation of the drug inhalation. The drug combination of isoflurane and nitrous oxide appears to be a promising candidate for conscious sedation procedures, although its analgesic and mood-altering effects need to be studied more systematically. PMID- 8712394 TI - Oral transmucosal fentanyl citrate for premedication in adults. AB - This study was designed to assess the efficacy of oral transmucosal fentanyl citrate (OTFC) for premedication in an adult population and to determine its effects on anxiety, sedation, gastric volume, and gastric fluid acidity. The fentanyl citrate is incorporated in a lozenge mounted on a handle (oralet). The effects of OTFC, placebo oralet, and no premedication were compared in a prospective, double-blind study on 90 adult ASA physical status I and II patients undergoing same-day admission surgery. Patients were randomly assigned to one of three groups: OTFC group (n = 30), placebo group (n = 30), and control group (n = 30). Arterial blood pressure, heart rate, respiratory frequency, and oxygen saturation determined by pulse oximetry were recorded before any premedication was given, and then every 10 min until the patient was taken to the operating room. Baseline anxiety and sedation levels were assessed to ensure group similarity immediately before premedication was given and at the more anxiety provoking phase upon entering the operating room. Anxiety levels were rated using the Spielberger State-Trait Anxiety Inventory short form and sedation levels were assessed with the Ramsay scale. Side effects, as reported by the patients, were also recorded. Gastric contents were aspirated via an orogastric tube after induction of anesthesia and were measured for volume and pH. No significant differences were found among the three groups in mean arterial pressure, heart rate, or respiratory frequency. Initial oxygen saturation levels in all groups decreased after 30 min but not less than 96% except for one patient in the OTFC group, who decreased to 88%. On entering the operating room, the OTFC group demonstrated significantly higher levels of anxiolysis than the control group, but no significant differences were seen between the OTFC and the placebo groups or the placebo and control groups. Mean gastric volumes (OTFC, 29 mL; placebo, 26 mL; control, 24 mL) and pH (OTFC, 2.0; placebo, 1.8; control, 2.1) were similar in all groups. There were no significant differences among the groups in levels of sedation achieved. Mild dizziness or light-headedness was the most commonly reported side effect in 23% of the OTFC group. In the OTFC group, 71.4% like the premedicant effect as compared to 46.4% of the placebo group. Most of the groups found the oralet method of premedicant delivery very acceptable. This study demonstrates that the OTFC oralet is an effective anxiolytic in adults. It has minimal side effects and is prepared in an acceptable format. There was no measurable increase in gastric contents or acidity in the oralet groups, compared to those patients who were given nothing by mouth. PMID- 8712395 TI - Alfentanil modifies the neurocirculatory responses to desflurane. AB - Activation of the sympathetic nervous system occurs in response to desflurane, causing tachycardia and hypertension. Fentanyl partially blunts the hemodynamic effects of desflurane but fails to attenuate the sympathetic response. This study determined the clinical effectiveness and dose response of alfentanil on the neurocirculatory responses to desflurane. Twenty-five healthy, male volunteers were randomized into one of three groups to receive either placebo (n = 9), 10 micrograms/kg intravenous (IV) bolus alfentanil (n = 9), or 20 micrograms/kg IV bolus alfentanil (n = 7) in conjunction with anesthetic induction by propofol, 2.5 mg/kg. Mean arterial pressure (MAP, radial artery), heart rate (HR), and efferent muscle sympathetic nerve activity (SNA, peroneal nerve) were recorded. After conscious baseline measurements, anesthesia was induced by propofol and alfentanil/placebo. One minute later, the desflurane vaporizer was activated at 11%. Neurocirculatory measurements were recorded for 11 min. There were no differences between the groups at conscious baseline. Induction of anesthesia was associated with significantly decreased MAP in the placebo and the 10 micrograms/kg alfentanil groups and increased HR in all groups with little change in SNA. In placebo subjects, desflurane administration increased HR and MAP above baseline. In both alfentanil groups, during desflurane administration HR and MAP never increased significantly above baseline. However, SNA was significantly increased in both groups. Alfentanil effectively blunts the hemodynamic changes but not the sympathetic responses associated with rapid increases in the inspired concentration of desflurane. PMID- 8712396 TI - Identification of human liver cytochrome P-450 3A4 as the enzyme responsible for fentanyl and sufentanil N-dealkylation. AB - Alfentanil, sufentanil, and fentanyl are synthetic opioids that are metabolized by oxidative N-dealkylation in the liver. We have previously shown that cytochrome P-450 3A4 (CYP3A4) contributes significantly to human liver microsomal alfentanil oxidation. Since identification of specific drug-metabolizing enzymes allows prediction of the variables affecting drug metabolism, the purpose of the present study was to identify the P-450 enzymes responsible for sufentanil and fentanyl metabolism in human liver microsomes. Microsomal preparations fortified with a reduced nicotinamide-adenine dinucleotide phosphate-generating system were incubated with 0.25 microM 3H-fentanyl or 3H-sufentanil. Rates of N-dealkylated metabolite formation significantly correlated with nifedipine oxidation activity (a marker of CYP3A4 activity) for fentanyl and sufentanil (r = 0.93 and 0.87, n = 18, respectively), but not with the oxidation activity for ethoxyresorufin (CYP1A2), S-mephenytoin (CYP2C19), bufuralol (CYP2D6), or chlorzoxazone (CYP2E1). Gestodene and troleandomycin (chemical inhibitors of CYP3A4) and antibody to CYP3A4 inhibited N-dealkylation of fentanyl and sufentanil. Chemical inhibitors of CYP2C, 2E1, and 2D6 did not inhibit N-dealkylation of fentanyl and sufentanil. Recombinant CYP3A4 expressed in Escherichia coli showed N-dealkylation activity of fentanyl and sufentanil, while expressed CYP1A2, 2C10, and 2E1 enzymes did not. We conclude that CYP3A4 is responsible for fentanyl and sufentanil N dealkylation in vitro. PMID- 8712397 TI - Renal and hepatic function in surgical patients after low-flow sevoflurane or isoflurane anesthesia. AB - The safety of low-flow sevoflurane anesthesia, which produces higher concentrations of toxic compounds, has been questioned. One hundred surgical patients received sevoflurane or isoflurane anesthesia at a total flow rate of 1 L/min. End-tidal CO2 concentrations and inspired and end-tidal anesthetic concentrations were monitored during anesthesia. Pre- and postanesthetic clinical laboratory studies were performed in both groups, and no significant differences were found between groups. In the sevoflurane group, the concentrations of degradation products in the circuit were measured by gas chromatography and the temperature of the CO2 absorbent was also measured. Two degradation products were detected: CF2 = C(CF3-O-CH2F (Compound A) and CH3OCF2CH(CF3)OCH2F (Compound B). The highest measured mean concentration of Compound A was 24.6 +/- 7.2 (13.6 41.3) ppm, and that of Compound B (detected in 12 patients) was 1.5 ppm. In both groups, total bilirubin, direct bilirubin, aspartate aminotransferase, and alanine aminotransferase were increased postoperatively. There was no difference between groups. Low concentrations of Compound A were present in low-flow sevoflurane anesthesia, but no significant differences in clinical laboratory values were observed between low-flow sevoflurane and isoflurane anesthesia. PMID- 8712398 TI - An evaluation of the antiinflammatory effects of intraarticular synthetic beta endorphin in the canine model. AB - We sought to evaluate the antiinflammatory effects of synthetic human beta endorphin (SHB) when injected into the canine knee joint. Sixteen healthy dogs, aged 1-2 yr, were selected. SHB was injected pre- and postinjury into each knee. The sample size was n = 32 after a randomized factorial arrangement; 2 x 4 with four cases per treatment being performed. Factors considered were: Factor A with two levels: A1 = Preinjury and A2 = Postinjury; Factor B (SHB dose) with four levels: B1 = Control, B2 = 250 micrograms, B3 = 500 micrograms, B4 = 1000 micrograms. The control group received 0.9% NaCl solution. Anesthesia was induced with intravenous thiopental, 14 mg/kg, and acepromazine, 0.5 mg/kg. Injury was produced with an intraarticular injection of 4 mL HCl 0.5 M, which was left in situ for 20 min. Inflammation was measured using the 610 nm absorbency of Evans blue extravasate in biopsy specimens. Histopathologic studies were performed on each knee. We found that beta-endorphin has a clear, dose-related, antiinflammatory effect, reducing the tissue extravasation of Evans blue and its absorbency, especially with large doses. This finding was consistent with the histopathologic observations. We conclude that SHB has an antiinflammatory effect. It is still not clear which mechanisms inhibit polymorphonuclear cell adhesion to vascular endothelium or cell and plasmatic protein extravasation. PMID- 8712399 TI - Motor and sensory disability has a strong relationship to induction dose of thiopental in patients with the hypertropic variety of Charcot-Marie-Tooth syndrome. AB - In a prospective study, we determined the anesthetic induction dose of thiopental and the clinical variables influencing the appropriate induction dose of thiopental in 20 patients with the hypertrophic variety of Charcot-Marie-Tooth syndrome (CMT). As controls we chose 50 patients without CMT. Motor disturbance was evaluated in terms of muscle weakness of the distal lower and upper extremities. We examined sensory disturbance by evaluating loss of sensation in the index finger and great toe. The preinduction cardiac output was measured by echocardiography. Anesthesia was induced with repeated injections of 50 mg thiopental. The minimum induction dose of thiopental (MID) was confirmed when the eyelash reflex ceased. We maintained anesthesia with enflurane and nitrous oxide. The 95% confidence interval of the MID in patients used as the controls was 2.5 4.9 mg/kg. The MID in 11 patients with CMT was less than 2.5 mg/kg. MIDs in the patients with CMT were significantly smaller than those of the control patients (P < 0.0001). Also we found a strong relationship between the MID and the severity of both motor and sensory disturbances (P = 0.003 and 0.002, respectively). There was no relationship between the MID and other clinical variables, such as age, gender, inherited type, body weight, and preinduction cardiac output. Because delay in the recovery from anesthesia can be caused by an inappropriate dose of thiopental in CMT patients in whom motor and sensory function is seriously impaired, the dose of thiopental probably should be reduced and based on the individual patient's response. PMID- 8712400 TI - Blood transfusion-induced immunomodulation. PMID- 8712401 TI - Chronic diarrhea: an uncommon side effect of celiac plexus block. PMID- 8712402 TI - The use of heliox and the laryngeal mask airway in a child with an anterior mediastinal mass. PMID- 8712403 TI - Systemic beta-adrenoceptor function and ophthalmic beta-adrenergic blockers. PMID- 8712404 TI - Spinal anesthesia after failed epidural anesthesia. PMID- 8712405 TI - Gastroesophageal reflux with the laryngeal mask. PMID- 8712406 TI - Complication of catheter knotting after right cephalic vein cannulation. PMID- 8712407 TI - Combined use of indirect orotracheal intubation techniques. PMID- 8712408 TI - Avoid rubber fingers. PMID- 8712409 TI - An unusual complication of epidural analgesia in a morbidly obese parturient. PMID- 8712410 TI - 1 + (-1) = 0, or not all anesthetic sites are created equal. PMID- 8712411 TI - Blocks to facilitate lithotomy positioning in spastic paraplegics. PMID- 8712412 TI - Patient comfort: spinal versus epidural anesthesia for cesarean section. PMID- 8712413 TI - Propofol induction is more effective than ondansetron in prophylaxis of postoperative nausea, but not vomiting. PMID- 8712414 TI - Epidural test dosing. PMID- 8712415 TI - Isoflurane-induced coronary vasodilation is preserved in reperfused myocardium. AB - Isoflurane causes vasodilation in the coronary circulation. The present study evaluated whether this action is preserved after a brief coronary occlusion followed by reperfusion. Fourteen open-chest dogs anesthetized with fentanyl and midazolam were studied. The left anterior descending coronary artery was perfused via an extracorporeal system with normal arterial blood or with arterial blood equilibrated with 1.4% (1 minimum alveolar anesthetic concentration [MAC]) isoflurane. Coronary perfusion pressure was maintained at 90 mm Hg. Coronary blood flow (CBF) was measured with a Doppler flow transducer. Steady-state changes in CBF during isoflurane, and during intracoronary infusions of acetylcholine (Ach; 20 micrograms/min), an endothelium-dependent vasodilator, and sodium nitroprusside (SNP; 80 micrograms/min), an endothelium-independent vasodilator, were compared in normal myocardium and in myocardium subjected to 15 min of ischemia (due to cessation of perfusion) followed by 30 min of reperfusion. Ischemia-reperfusion had no significant effect on the increases in CBF by isoflurane (421% +/- 88% vs 388% +/- 84%) or SNP (115% +/- 18% vs 135% +/- 19%), whereas it attenuated these increases in CBF by Ach (232% +/- 38% vs 143% +/- 21%). In conclusion, a brief period of myocardial ischemia followed by reperfusion did not affect the coronary vasodilating effects of isoflurane and SNP, although it blunted these effects of Ach. The present findings provide further evidence suggesting that the ability of isoflurane to relax coronary vascular smooth muscle is independent of the nitric oxide-cyclic guanosine monophosphate pathway. PMID- 8712416 TI - Don't abandon test doses. PMID- 8712417 TI - Research activity among Japanese anesthetists. PMID- 8712418 TI - When not to give a test dose: one more reason? PMID- 8712419 TI - Breathe in, breathe out. PMID- 8712420 TI - Use of a fiberoptic bronchoscope adapter during jet ventilation. PMID- 8712421 TI - Protamine and acute depletion of magnesium limit bone response to parathyroid hormone. AB - The effect of protamine on calcium homeostasis was studied in nine pediatric patients undergoing cardiopulmonary bypass. Total serum calcium decreased from 8.44 mg/dL to 7.49 mg/dL (P < 0.05) after protamine. Ionized calcium decreased from 1.39 to 1.31 mmol/L (P < 0.05). A bioassay determined the etiology of this response. Bone disks were placed in sera, protamine, parathyroid hormone, parathyroid hormone antibody, or magnesium-depleted solutions, then were incubated in solutions with known calcium content. The change in the media's calcium concentration reflects the bone's response to the initial stimulus. Calcium change is expressed as Experimental delta/Control delta (E/C). Normal bone responds to parathyroid hormone, E/C = 0.59 (P < 0.001). Protamine-treated bone loses this response, E/C = 0.9 (P = not significant [NS]). A parathyroid hormone-induced osteoblast messenger was found. Protamine-treated bone continued to respond to this messenger, E/C = 0.42 (P < 0.001). Bone showed reversible loss of response to parathyroid hormone after incubation in magnesium-free solution, E/C = 0.93 (P = NS). With reincubation in magnesium, E/C = 0.69 (P < 0.01). Since protamine blocks parathyroid receptors, and magnesium depletion limits the bone's response to parathyroid hormone, this may explain the persistent hypocalcemia seen in some patients undergoing cardiopulmonary bypass. PMID- 8712422 TI - Positive pressure ventilation with the laryngeal mask airway in children. AB - We studied the safety of positive pressure ventilation (PPV) when using the size 2 laryngeal mask airway (LMA) in 46 ASA physical status I or II children (aged 38 +/- 21 mo) undergoing elective surgery. The LMA cuff was inflated in incremental steps to achieve a cuff leak pressure > or = 15 cm H2O. Abdominal circumference was measured before and after PPV in study patients, as well as in a control group managed with tracheal intubation. Cuff leak pressure was 17 +/- 4 cm H2O (range 12-34 cm H2O). Forty-five patients successfully underwent PPV. Gas leak around the LMA cuff prevented PPV in one infant. The only respiratory variable that changed significantly was end-tidal CO2, which decreased from 40 +/- 6 to 34 +/- 5 mm Hg. Abdominal circumference increased in 28 patients but was not associated with any complications. Change in abdominal circumference in the study group was not significantly different from that observed in the control group. However, abdominal circumference increased 8 cm in one study patient, prompting insertion of an orogastric tube. The size 2 LMA provides an effective airway for PPV. Mild gastric distention often occurs. The risk of clinically significant gastric distention appears to be small, but it warrants close monitoring. We conclude that with certain precautions described in the text, the size 2 LMA provides a relatively safe airway for PPV in children. PMID- 8712423 TI - Does age or pseudocholinesterase activity predict mivacurium infusion rate in children? AB - Previous studies have suggested that the mivacurium infusion rate to maintain target twitch depression is greater in children than in adults, and that there is only a limited relationship between pseudocholinesterase activity and mivacurium infusion rate in children. We therefore examined whether mivacurium infusion rates are larger in children than in adults, and whether pseudocholinesterase activity influences mivacurium infusion rate in children. In 20 children aged 1-9 yr, mechanical twitch response to ulnar nerve train-of-four stimulation was measured; concurrent data were obtained in 14 adults aged 18-58 yr. All patients were anesthetized with N2O and isoflurane, 0.75 minimum alveolar anesthetic concentration (MAC) (age-adjusted). Mivacurium was infused at constant rates for > 15 min targeting 50% and 90% twitch depression. The Hill equation was fit to the resulting values for twitch depression versus mivacurium infusion rate to predict infusion rates producing 50% and 90% twitch depression (IR50 and IR90, respectively). The relationship of IR50 and IR90 to pseudocholinesterase activity was determined by linear regression; values for children and adults were compared by analysis of covariance. For children, IR50 (r2 = 0.22, P = 0.038) but not IR90 (r2 = 0.11 P = 0.21) was related to pseudocholinesterase activity. Infusion rates were approximately twice as large in children as in adults. We confirm that mivacurium infusion rates are larger in children than in adults and demonstrate a relationship between pseudocholinesterase activity and infusion rates. PMID- 8712424 TI - Preoperative pregnancy testing in adolescents. AB - Anesthetics and other drugs used during the perioperative period may have teratogenic or abortive effects. The pregnancy status of surgical patients is often unknown. This investigation examined retrospectively the results of 2 yr of mandatory pregnancy testing in 412 adolescent surgical patients. The overall incidence of positive tests was 1.2%. Five of 207 patients aged 15 yr and older tested positive, for an incidence of 2.4% in that group. None of the 205 patients under the age of 15 yr had a positive pregnancy test. We conclude that mandatory pregnancy testing is advisable in adolescent surgical patients aged 15 yr and older. PMID- 8712425 TI - Preoperative multimodal analgesia facilitates recovery after ambulatory laparoscopic cholecystectomy. AB - Laparoscopy approach to cholecystectomy has shortened the recovery period, reducing discharge times from 1 to 3 days to same-day discharge. We hypothesize that the use of more than one modality to prevent postoperative pain may be more efficacious than single modality. Patients were randomized to a treatment (n = 24) or control (n = 25) group and studied using a prospective, double-blind design. Preoperatively, at 45 min before induction of anesthesia, the treatment group received an intramuscular (IM) bolus injection of meperidine 0.6 mg/kg and ketorolac 0.5 mg/kg. The control group received two bolus IM injections of placebo (normal saline). Ten minutes before incision, local anesthesia (treatment group) or saline (control group) was infiltrated into the skin of each patient. Anesthetic management, postoperative pain, and nausea treatment were standardized. Pain and nausea assessment were done 1 h preoperatively, 0, 0.5, 1, 2, 3, and 4 h postoperatively, at discharge, and 10, 24, and 48 h postoperatively. Patients were discharged by scoring criteria. Postoperatively, significantly more patients in the treatment group were without pain on arrival in the postanesthesia care unit (PACU), 12/21 (57.1%) vs 1/24 (4.2%) in the control group (P < 0.001). Similarly, the severity of pain was sixfold less in the treatment group than in the control group. The incidence of nausea in the PACU was significantly less in the treatment group; 4.7% vs 29.5% in the control group (P < 0.05). Patients from the treatment group satisfied Postanesthesia Discharge Score significantly earlier than those in the control group (281 +/- 12 min vs 375 +/- 19 min; P < 005). The concomitant use of local anesthetic and nonsteroidal antiinflammatory and opioid drugs proved to be highly effective in our patients, resulting in faster recovery and discharge. PMID- 8712426 TI - Coupling between local cerebral blood flow and metabolism after hypertonic/hyperoncotic fluid resuscitation from hemorrhage in conscious rats. AB - The effects of small volume hypertonic/hyperoncotic fluid resuscitation from hemorrhage on brain metabolism and blood flow were evaluated by autoradiographic techniques with high spatial resolution. The data were compared to fluid resuscitation with a volume equal to shed blood of isotonic 6% hydroxyethyl starch solution (HES) and a control group without hemorrhage and fluid resuscitation (n = 6 in each group). In conscious rats, volume-controlled hemorrhage for 30 min (30 mL/kg body weight, resulting in a blood loss of approximately 50% of the circulating blood volume) was followed by intravenous infusion of a hypertonic/hyperoncotic saline hydroxyethyl starch solution (HTHO; 7.5% saline/10% hydroxyethyl starch, 4.0 mL/kg body weight). Local cerebral blood flow (LCBF) and local cerebral glucose utilization (LCGU) were measured in 34 brain structures 2 h after fluid resuscitation by means of the quantitative autoradiographic iodo [14C]antipyrine and 2-[14C]-deoxy-D-glucose methods. Compared to the untreated control group, LCBF increased significantly in all brain regions analyzed after fluid resuscitation with HTHO (mean, +63%) or HES (mean, +56%). The increases in LCBF after fluid resuscitation were sufficient to restore cerebral oxygen delivery to the level calculated for the untreated control group. LCGU was unchanged after fluid resuscitation. The close relationship between LCGU and LCBF observed in the control group (r = 0.95) was preserved after hemorrhage and fluid resuscitation with HTHO (r = 0.97) and HES (r = 0.96), although the LCBF-to-LCGU ratio was reset to a higher level (1.5 mL/mumol in the control group and 2.7 mL/mumol after fluid resuscitation with HTHO and HES, P < 0.05). We conclude that the increase in LCBF compensates for the reduction of arterial oxygen content to maintain cerebral oxygen delivery. Therefore, "small volume resuscitation" appears to be as effective as resuscitation with large volumes of isotonic HES in meeting the circulatory and metabolic demands of the brain tissue within the first 2 h after fluid resuscitation from hemorrhage. PMID- 8712427 TI - Marked hemodilution increases neurologic injury after focal cerebral ischemia in rabbits. AB - Moderate hemodilution (hematocrit approximately 30%) reduces neurologic injury after focal cerebral ischemia. In contrast, both clinical and experimental studies suggest that marked hemodilution (hematocrit < 30%) may exacerbate neurologic injury. We compared the effect of marked versus minimal hemodilution on cerebral infarct volume after focal cerebral ischemia in rabbits. Anesthetized New Zealand White rabbits underwent hemodilution by exchange of arterial blood with 6% high molecular weight hydroxyethyl starch. In the marked hemodilution group (n = 15) the target hemoglobin concentration was 6 g/100 mL. In the minimal hemodilution group (n = 15) the target hemoglobin concentration was 11 g/100 mL. After hemodilution, middle cerebral artery occlusion was achieved by embolizing an autologous blood clot via the internal carotid artery. Four hours after embolization, the animals were killed and their brains removed. Brains were sectioned, stained with 2,3,5-triphenyltetrazolium chloride, and infarct volumes determined via quantitative image analysis. Systemic physiologic variables were similar between groups, except for arterial hemoglobin concentration. The percentage of hemispheric infarct was significantly larger in the marked hemodilution group as compared to the minimal hemodilution group, 70% +/- 19% vs 51% +/- 23%, respectively (mean +/- SD); P = 0.02. Similarly, the percentage of infarct was greater in the hemodilution group as compared to the minimal hemodilution group in both cortex (73% +/- 18% vs 54% +/- 23%, respectively; P = 0.02) and subcortex (62% +/- 25% vs 44% +/- 23%, respectively; P = 0.04). These findings indicate that marked hemodilution exacerbates neurologic injury resulting from permanent focal ischemia. Although some degree of hemodilution may improve neurologic outcome, the advantage is lost at an extreme level of therapy. PMID- 8712428 TI - Trimethaphan versus sodium nitroprusside for the control of proximal hypertension during thoracic aortic cross-clamping: the effects on spinal cord ischemia. AB - Sodium nitroprusside (SNP) has been used to control the proximal hypertension associated with thoracic aortic cross-clamping (TACC) during thoracic aortic surgery. It worsens neurologic outcome, presumably by further decreasing distal arterial pressure and increasing cerebrospinal fluid (CSF) pressure, thereby worsening the spinal cord perfusion pressure (SCPP). Trimethaphan does not increase CSF pressure. Therefore, the present study investigates the effect of trimethaphan versus SNP to control proximal hypertension during TACC on neurologic outcome. Two groups, each with eight mongrel dogs, were studied. All animals underwent descending TACC for 45 min. The mean proximal aortic blood pressure was maintained at 95-100 mm Hg by the use of SNP or trimethaphan. Distal aortic pressure was allowed to vary. The dogs were neurologically evaluated 24 and 48 h later by a blinded observer. During cross-clamping, there was no difference in mean proximal aortic pressure between groups. After 10 min of cross clamping, distal aortic pressure was higher (P < 0.01), CSF pressure was lower (P < 0.01), and SCPP was higher (P < 0.005) in the trimethaphan group as compared with the SNP group (group effect). Neurologic outcome as assessed by Tarlov's score was better at 24 and 48 h in the trimethaphan group (P < 0.05). Histopathologic injury trended with worsened neurologic outcome. We conclude that 1) trimethaphan produced higher SCPP than SNP, and 2) neurologic outcome was better in the trimethaphan group. PMID- 8712429 TI - Comparison of the effects of enflurane/N2O on the 40-Hz auditory steady-state response versus the auditory middle-latency response. AB - The auditory middle-latency response (AMLR) is a sequence of negative-positive waves occurring 12-50 ms after the onset of an auditory stimulus presented at rates of 10/s or less. When the rate of stimulus presentation is increased to approximately 40/s, overlapping of the AM-LRs results in a sustained, nearly sinusoidal wave, called the "40-Hz auditory steady-state response" (40-Hz ASSR). The AMLR and 40-Hz ASSR have been used to study the effects of general anesthetics on the brain. The primary aim of this investigation was to determine whether the effects of a general anesthetic, namely enflurane, on the 40-Hz ASSR can be predicted from its effects on the AMLR. A secondary aim was to examine the relationship between the level of consciousness and the 40-Hz ASSR during emergence from anesthesia. Twelve ASA class I-II women undergoing reduction mammoplasty were tested. Anesthesia was induced with fentanyl (3 micrograms/kg) and thiopental (3-5 mg/kg) intravenously and was maintained with enflurane (0.5%, 0.8%, or 1.1% end-tidal; four patients per concentration; random assignment) in N2O (66% end-tidal), along with fentanyl (1 microgram/kg as needed). The 40-Hz ASSR and AMLR were recorded before induction and during anesthesia and surgery. The 40-Hz ASSR was also recorded during emergence. The amplitude of the 40-Hz ASSR was reduced profoundly during anesthesia and surgery (P < 0.001). The attenuation was not dose-dependent, and was much more pronounced than predicted by the effects of enflurane on the AMLR. The 40-Hz ASSR during anesthesia was surprisingly large (0.09 and 0.11 microV) in two patients, both of the 1.1% enflurane group. The regaining of the ability to follow verbal commands was associated with a significant (P < 0.001) increase in the amplitude of the 40-Hz ASSR. We conclude that, although auditory neurons remain capable of responding at a slow stimulus rate during enflurane-N2O anesthesia, their ability to be driven at a faster stimulus rate is markedly curtailed. The 40-Hz ASSR may be useful for detecting unintentional intraoperative awareness because the return of consciousness is associated with a clear increase in amplitude. PMID- 8712430 TI - Ilioinguinal iliohypogastric nerve blocks--before or after cesarean delivery under spinal anesthesia? AB - Preoperative local anesthetic blockade of somatosensory pathways involved with skin incision and other noxious perioperative stimuli may "preempt" or attenuate the postoperative pain response. Since the Pfannenstiel incision lies within the L1 dermatome, bilateral ilioinguinal, iliohypogastric nerve blocks (IINBs) should provide analgesia after low transverse cesarean section. We designed this study to compare the analgesic effect of IINBs placed before or after cesarean delivery. Forty-six patients undergoing cesarean delivery with spinal anesthesia were enrolled. Patients were randomly assigned to one of three groups: Before, After, or None. Bilateral IINBs were placed with 0.5% bupivacaine, 10 mL to each side. Twenty-two patients had IINBs placed before surgery (11 failed blocks), 12 had blocks placed after surgery, and 12 had no block. Morphine, through a patient controlled analgesia (PCA) pump, provided additional postoperative analgesia. A blinded observer assessed 24-h morphine use as well as patient satisfaction and pain scores ("incisional" and "overall") for 96 h. Although there were no consistent differences in pain scores among the groups, patients in the After group occasionally reported more pain than those in the Before and None groups. Patient satisfaction and morphine use did not differ among the groups. We conclude that there is no benefit to ilioinguinal nerve blocks, either before or after surgery, in patients who receive spinal anesthesia for elective cesarean delivery. Our finding of increased pain in the After group is perplexing and requires confirmation. PMID- 8712431 TI - Coronary perfusion pressure during cardiopulmonary resuscitation after spinal anesthesia in dogs. AB - Cardiac arrest during spinal anesthesia is a rare event, but when it does happen cardiopulmonary resuscitation (CPR) is often ineffectual. This study examines the effect of spinal anesthesia on coronary perfusion pressure (CPP) during CPR and the subsequent response of CPP to epinephrine administration. Twenty mongrel dogs were anesthetized, and randomly assigned to a spinal injection with either 0.5 mg/kg bupivacaine or with an equivalent volume of normal saline. Twenty minutes later, ventricular fibrillation was electrically induced and after 1 min CPR was started. CPP was measured every minute. After 4 min of CPR, epinephrine 0.01 mg/kg was given followed by 0.1, 0.2, and 0.4 mg/kg epinephrine intravenously (IV) at 6, 8, 10 min of CPR, respectively. The bupivacaine (n = 11) group had significantly less CPP than the sham spinal (n = 8) group, 12-13 mm Hg as compared to 27-34 mm Hg. Only 4/11 dogs (36%) in the bupivacaine group had CPP > or = 15 mm Hg during the first 4 min after arrest as compared to 8/8 (100%) in the sham spinal group. This increased to 7/11 dogs (64%) after 0.01 mg/kg epinephrine and to 9/11 after 0.1 mg/kg epinephrine. Total spinal anesthesia decreases CPP and thus the efficacy of CPR in dogs below the threshold previously established for predicting successful resuscitation. Epinephrine is effective in increasing CPP during CPR above the critical threshold. These data suggest that if cardiac arrest occurs during spinal anesthesia, epinephrine should be given in doses of 0.01-0.02 mg/kg IV initially and then increasing to 0.1 mg/kg IV. When this does not work, and ineffective CPR is suspected, alternative resuscitative measures should be considered. PMID- 8712432 TI - Epiduroscopic changes in patients undergoing single and repeated epidural injections. AB - Using a superfine fiberscope with an outer diameter of 0.75 mm, the effect of repeated injections through a single epidural catheter on the epidural space was examined in 18 patients. The subjects were divided into a control group of 10 patients who had not previously received epidural anesthesia (EA) and an EA group of 8 patients who had received repeated EA with 4-6 mL 0.25% bupivacaine through an epidural catheter, two to three times per day for 7-14 days. The epidural space was observed through a fiberscope passed through a 17-gauge Tuohy needle. The epidural space was occupied by large masses of fat, and the blood vessels and connective tissue were confirmed. Adverse reactions in the epidural tissue, such as hemorrhage or congestion with engorgement, were observed in four patients in the EA group. Five patients in the EA group experienced pain when the fiberscope was inserted into the epidural space. These investigations show that continuous EA might be followed by a high incidence of nonspecific epidural changes. Superfine fiberscope may be useful in the detection or diagnosis of local epidural reaction. PMID- 8712433 TI - The analgesic response to intravenous lidocaine in the treatment of neuropathic pain. AB - This study was performed in order to determine concentration-effect, and graded and quantal dose-response relationships for the clinical administration of intravenous (IV) lidocaine to patients with neuropathic pain. Thirteen patients were administered 500 mg of IV lidocaine at a rate of 8.35 mg/min over 60 min. Visual analog pain scores and venous blood samples were obtained concomitantly at 10 min intervals for 60 min. Blood samples were also obtained for determination of serum and serum water lidocaine concentrations at the onset of analgesia and at the time complete pain relief was attained. Lidocaine concentrations were determined by gas chromatography. Graded dose-response curves were prepared individually and for the group as a whole, and a quantal dose-response curve was prepared for the entire group. The dose-response relationship for IV lidocaine was characterized by large increases in pain relief for concomitant minimal increases in dosage. The difference between the ED50 (372.0 mg) and the ED90 (416.5 mg) was 44.5 mg of lidocaine (5.3 min of infusion). The concentration effect relationship was also steep with pain scores abruptly decreasing over a range of 0.62 microgram/mL of lidocaine. Interestingly, the free concentration of lidocaine had no better correlation with the onset of analgesia or the attainment of complete analgesia than the serum concentration of lidocaine. This suggests that the mechanism of analgesia to IV lidocaine may not be based upon a conventional concentration-effect relationship. In conclusion, the results of this study suggest that the analgesic response to IV lidocaine is best characterized by a precipitous "break in pain" over a narrow dosage and concentration range. PMID- 8712434 TI - The effects of electrical stimulation at different frequencies on perception and pain in human volunteers: epidural versus intravenous administration of fentanyl. AB - The study was performed to determine whether epidural fentanyl produced segmental sensory changes to electrical stimulation at different frequencies. Eight healthy volunteers received fentanyl 1 microgram/kg both intravenously and epidurally in a randomized, double-blind, cross-over fashion. Perception thresholds and amount of current required to elicit a predetermined level of moderate pain (Cmp) at 5,250, and 2000 Hz stimulation were measured at ipsilateral dermatomes C2 and L2 at 0, 5, 15, 30, 45, and 60 min after injection. Perceptions to 5,250, and 2000 Hz stimulation were unaffected by either intravenous or epidural fentanyl (P > 0.08). Intravenous fentanyl increased Cmp at both 5 and 250 Hz at both dermatomes (P < 0.004) and thus did not produce segmental analgesia. In contrast, epidural fentanyl increased Cmp only at the L2 dermatome and only at 5 Hz (P = 0.005). We conclude that an epidural bolus of fentanyl results in segmental spinal analgesia to transcutaneous electrical stimulation only at specific frequencies. Furthermore, pain produced by stimulation at 5 Hz may have a different pharmacology than pain produced by 250 Hz stimulation. PMID- 8712435 TI - Intramuscular rocuronium in infants and children--is there a need? PMID- 8712436 TI - Intramuscular rocuronium in infants and children. Dose-ranging and tracheal intubating conditions. AB - BACKGROUND: Rocuronium's rapid onset and intermediate duration of action with intravenous administration suggests that intramuscular administration might facilitate tracheal intubation without producing prolonged paralysis. Accordingly, in infants and children, the authors measured onset at the adductor pollicis and respiratory muscles to determine the optimal dose (phase I), then gave this optimal dose to determine the optimal time for tracheal intubation (phase II). METHODS: The authors studied 45 unpremedicated patients aged 3 months to 5 yr. In phase I, 25 patients were anesthetized with nitrous oxide and halothane and breathed spontaneously; twitch tension and minute ventilation were measured. Rocuronium (800-2,400 micrograms/kg) was injected into the quadriceps or deltoid muscle; doses varied, using an "up-down" technique, the goal being to bracket the dose depressing twitch 75-90% within 5 min. In phase II, deltoid injections of the optimal dose from phase I (infants: 1,000 micrograms/kg; children: 1,800 micrograms/kg) were given to 20 patients anesthetized with 0.82 1.0% halothane. Tracheal intubation was attempted 1.5-3.0 min later; time to tracheal intubation was varied, using an "up-down" technique. RESULTS: In phase I, 5 of 7 patients given quadriceps injections (1,200-2,200 micrograms/kg) had slow onset of twitch and ventilatory depression. With deltoid injections (800 2,400 micrograms/kg), all patients developed complete twitch depression; median time to 50% depression of minute ventilation was 3.2 min in infants and 2.8 min in children. In phase II, intubating conditions were consistently adequate or good-excellent at 2.5 min in infants and 3.0 min in children. Initial twitch recovery was at 57 +/- 13 min (mean +/- SD) in infants and 70 +/- 23 min in children. CONCLUSIONS: Deltoid injections of rocuronium, 1,000 micrograms/kg in infants and 1,800 micrograms/kg in children, rapidly permit tracheal intubation in infants and children, despite a light plane of anesthesia. Duration of action of these large doses might limit clinical utility. PMID- 8712437 TI - Isoflurane produces marked and nonlinear decreases in the vasoconstriction and shivering thresholds. AB - BACKGROUND: Desflurane decreases the vasoconstriction and shivering thresholds disproportionately at high anesthetic concentrations. This result contrasts with the authors' previous report that isoflurane decreases the vasoconstriction threshold linearly. It is surprising that the basic shape of the concentration response curve should differ with these two otherwise similar anesthetics. Therefore, the hypothesis that isoflurane produces a nonlinear reduction in the vasoconstriction threshold was tested. Because the effect of isoflurane on shivering remains unknown, the extent to which isoflurane reduces the shivering threshold also was determined. METHODS: Eight men volunteered to be studied on four randomly ordered days: (1) a target end-tidal isoflurane concentration of 0.55%, (2) a target concentration of 0.7%, (3) control (no anesthesia) and a target end-tidal concentration of 0.85%, and (4) a target end-tidal concentration of 1.0%. Volunteers were surface-cooled until peripheral vasoconstriction and shivering were observed. We arithmetically compensated for changes in skin temperature using the established linear cutaneous contributions to control for each response. From the calculated thresholds (core temperatures triggering responses at a designated skin temperature of 34 degrees C), the concentration response relation was determined. RESULTS: Isoflurane administration produced a dose-dependent reduction in the vasoconstriction and shivering thresholds, decreasing each approximately 4.6 degrees C at an end-tidal concentration of 1%. Residual analysis indicated that the vasoconstriction and shivering thresholds were decreased in a nonlinear fashion during isoflurane administration. The vasoconstriction-to-shivering range was 1.5 +/- 0.8 degree C without isoflurane, and did not change significantly during isoflurane administration. CONCLUSIONS: The vasoconstriction-to-shivering range remained unchanged by isoflurane administration. In this regard, the effects of isoflurane are similar to those of desflurane, propofol, and alfentanil. The current data differ from the authors' previous report, in that the dose-dependence for vasoconstriction was nonlinear, with isoflurane reducing the threshold disproportionately at higher anesthetic concentrations. Differing dose-dependence in the two studies may result either because the current study's volunteers were not exposed to surgical stimulation and were given less isoflurane, or because of design limitations in the previous protocol. PMID- 8712438 TI - Efficacy of the self-inflating bulb in confirming tracheal intubation in the morbidly obese. AB - BACKGROUND: This study was designed to determine the incidence of false-negative and false-positive results when the self-inflating bulb (SIB) is used to differentiate tracheal from esophageal intubation in morbidly obese patients using two techniques. In technique 1, the SIB is compressed before it is connected to the tube; in technique 2, the SIB is compressed after connection to the tube. METHODS: With institutional review board approval, 54 consenting adult morbidly obese patients (body mass index > 35) undergoing elective surgical procedures were included in the study. After anesthetic induction and muscle relaxation, both the trachea and esophagus were intubated under direct vision with identical cuffed tubes. The efficacy of the SIB in verifying the position of both tubes was tested by a second anesthesiologist. The speed of reinflation was graded as rapid ( < 4 s) or none ( > 4 s), using both techniques. In the case of tracheal intubation, the absence of reinflation was recorded as a false-negative, whereas in cases of esophageal intubation, rapid reinflation was recorded as a false-positive. Identification of tube location by the second anesthesiologist was based on SIB reinflation results from techniques 1 and 2, as well as the presence of a flatuslike sound elicited by technique 2 in esophageally placed tubes. All patients were retested by the SIB after receiving three breaths of 400 500 ml each. In all patients exhibiting false-negative results, six obese patients exhibiting true-positive results, and four nonobese patients exhibiting true-positive results, tracheal responses to the SIB maneuvers were observed directly by a flexible fiberoptic bronchoscope incorporating an airtight system, 15-20 min after mechanical ventilation was instituted. RESULTS: The incidence of false-negative results was initially 30% with technique 1 and 11% with technique 2, but decreased to 4% when technique 2 was used after the delivery of three breaths. The second anesthesiologist initially identified tube location in 92.5% of patients correctly. After the delivery of three breaths, tube location was correctly identified in 96.3% of patients. Fiberoptic bronchoscopic examination of the patients exhibiting false-negative results revealed exaggerated inward bulging of the posterior tracheal membrane during reinflation of the SIB when technique 1 was used. CONCLUSIONS: Contrary to previous investigations in healthy patients, the current study demonstrates a high incidence of false-negative results when the SIB is used to confirm tracheal intubation in morbidly obese patients. If the SIB is used, the technique should include compression of the SIB after connection to the tube and should be used in conjunction with other clinical signs and technical aids. The mechanism of false-negative results in these patients seems to be related to reduction of caliber of airways secondary to a marked decrease in functional residual capacity, and collapse of large airways due to invagination of the posterior tracheal wall when sub-atmospheric pressure is generated by the SIB. PMID- 8712439 TI - Cardiac risk of noncardiac surgery in patients with asymmetric septal hypertrophy. AB - BACKGROUND: Many data are available regarding cardiac risk in patients with coronary artery disease undergoing noncardiac surgery, but few data are available regarding risk for patients with hypertrophic cardiomyopathy and asymmetric septal hypertrophy. METHODS: Seventy-seven patients with asymmetric septal hypertrophy were identified in whom an echocardiogram had been performed within 24 months of noncardiac surgery. Patients' charts were reviewed for data regarding surgical operations, including length of surgery, type of anesthesia, and intravascular monitoring used. Data regarding adverse perioperative cardiac events also were gathered. RESULTS: Forty percent (n = 31) of patients had one or more adverse perioperative cardiac events, including one patient who had a myocardial infarction and ventricular tachycardia that required emergent cardioversion. There were no perioperative deaths. All 31 patients had minor outcomes. Of the 77 patients, perioperative congestive heart failure developed in 12 (16%). Factors associated with adverse cardiac events were increasing length of surgical time (P < 0.01) major surgery (P < 0.05), and intensity of monitoring (P < 0.05). Age, gender, resting outflow tract gradient, systolic anterior motion of the anterior mitral leaflet, prior myocardial infarction, severity of mitral regurgitation, type of anesthetic, septal thickness, and the interval between echocardiogram and surgery were not associated with the occurrence of adverse cardiac events. CONCLUSION: Patients with asymmetric septal hypertrophy undergoing noncardiac surgery have a high incidence of adverse cardiac events, frequently manifested as congestive heart failure. However, irreversible cardiac morbidity and mortality was extremely low. Important independent risk factors for adverse outcome in all patients include major surgery and increasing duration of surgery. PMID- 8712440 TI - Changing anesthesiologists' practice patterns. Can it be done? AB - BACKGROUND: Because the ultimate purpose of new medical knowledge is to achieve improved health outcomes, physicians need to possess and use this knowledge in their practice. The authors introduced enhanced education and individualized feedback to reduce postoperative nausea and vomiting (PONV). The primary objective was to increase anesthesiologists' use of preventive measures to reduce PONV, and the secondary objective was to determine whether patient outcomes were improved. METHODS: After obtaining hospital ethics committee approval, the effect of education and feedback on anesthesiologist performance and the rate of PONV in major surgery elective inpatients during a 2-yr period was assessed. After baseline data collection (6 months), anesthesiologists at the study hospital received enhanced education (8 months) and individualized feedback (10 months). Parallel data collection was performed at a control hospital at which practice was continued as usual. The education promoted preventive measures (antiemetic premedication, nasogastric tubes, droperidol, metoclopramide). Individualized feedback provided the number of patients receiving promoted measures and the rate of PONV. The mean percentage of anesthesiologists' patients receiving at least one promoted measure and the rate of PONV were compared with baseline levels. RESULTS: At the study hospital, there was a significant increase in the mean percentage of the anesthesiologists' female patients receiving a preventive measure as well as a significant increase in the use of droperidol > or = 1 mg (P < 0.05) for all patients. The use of other promoted measures was unaffected. Absolute rates of PONV were unaffected at the study hospital until the post feedback period (decrease of 8.8% between baseline and post-feedback (P = 0.015)). CONCLUSION: It was demonstrated that enhanced education and individualized feedback can change anesthesiologists' practice patterns. The actual benefit to patients from use of preventive measures was limited when used in the everyday clinical situation. Therefore, only modest decreases in PONV were achieved, despite the use of preventive measures. PMID- 8712441 TI - Intravenous ondansetron in established postoperative emesis in children. S3A-381 Study Group. AB - BACKGROUND: In pediatric postsurgical patients, postoperative vomiting is a common occurrence that can delay recovery and result in unplanned hospital admissions after outpatient surgery. This randomized, double-blind, placebo controlled, multicenter study evaluated the efficacy and safety of ondansetron in the control of established postoperative emesis in outpatients aged 2-12 yr. METHODS: Screened for the study were 2,720 ASA physical status 1-3 children undergoing outpatient surgery during general anesthesia, which included nitrous oxide. Children experiencing two emetic episodes within 2 h of discontinuation of nitrous oxide were given intravenous ondansetron (n = 192; 0.1 mg/kg for children weighing < or = 40 kg; 4 mg for children weighing > 40 kg) or placebo (n = 183). RESULTS: The proportion of children with no emetic episodes and no use of rescue medication was significantly greater (P < 0.001) in the ondansetron group compared with placebo for both 2- and 24-h periods after study drug administration (78% of the ondansetron group and 34% of the placebo group for 2 h; 53% of the ondansetron group and 17% of the placebo group for 24 h). Among patients with at least one emetic episode or with rescue medication use, the median time to onset of emesis or rescue was 127 min in the ondansetron group compared with 58 min in the placebo group (P < 0.001). The median time from study drug administration until discharge was significantly shorter (P < 0.01) in the ondansetron group (153 min, range 44-593 min) compared with the placebo group (173 min, range 82-622 min). The incidence of potentially drug-related adverse events was similar in the ondansetron (3% of patients) and the placebo (4% of patients) groups. CONCLUSION: A single dose of ondansetron (0.1 mg/kg up to 4 mg) is effective and well tolerated in the prevention of further episodes of postoperative emesis in children after outpatient surgery. Administration of ondansetron also may result in a shorter time to discharge. PMID- 8712442 TI - Simulation of an epidural test dose with intravenous isoproterenol in awake and in halothane-anesthetized children. AB - BACKGROUND: An epidural test dose containing epinephrine does not reliably produce hemodynamic responses in children under halothane anesthesia. The purpose of this study was to determine hemodynamic responses to intravenous isoproterenol in both awake and halothane-anesthetized children. METHODS: After obtaining institutional review board approval and parental informed consent, 72 ASA physical status 1 or 2 children (2.8 +/- 1.7 yr) undergoing elective minor surgery were studied before and during anesthesia with 1.2 minimum alveolar concentration halothane. A bolus containing 0.25 mg/ kg bupivacaine and 0.05 microgram/kg, 0.075 microgram/kg, or 0.1 microgram/kg isoproterenol, or bupivacaine and saline was injected via a peripheral arm vein to simulate intravascular injection of an epidural test dose. RESULTS: Before induction of anesthesia, all patients showed a positive test response after isoproterenol injection (heart rate increase > or = 20 beats/min). During anesthesia, 79% of patients receiving 0.05 microgram/kg, 89% of patients receiving 0.075 microgram/kg, and 100% of patients receiving 0.1 microgram/kg met the criterion of a positive test response. Among each treatment group, all infants showed a positive test response. Blood pressure did not differ among the groups at any time. Transient benign dysrhythmias occurred in only one patient under halothane anesthesia receiving 0.075 microgram/kg isoproterenol. CONCLUSION: Isoproterenol at a dose of 0.1 microgram/kg is a sensitive indicator for intravascular injection of a test dose in children anesthetized with halothane and nitrous oxide. Isoproterenol at a dose of 0.05 microgram/kg approximates a minimal effective dose in awake children and in infants. After detailed studies on neural toxicity, isoproterenol could be of value as an epidural test agent in children. PMID- 8712443 TI - Thermoregulatory vasoconstriction does not impede core warming during cutaneous heating. AB - BACKGROUND: Although forced-air warming rapidly increases intraoperative core temperatures, it is reportedly ineffective postoperatively. A major difference between these two periods is that arteriovenous shunts are usually dilated during surgery, whereas vasoconstriction is uniform in hypothermic postoperative patients. Vasoconstriction may decrease efficacy of warming because its major physiologic purposes are to reduce cutaneous heat transfer and restrict heat transfer between the two thermal compartments. Accordingly, we tested the hypothesis that thermoregulatory vasoconstriction decreases cutaneous transfer of applied heat and restricts peripheral-to-core flow of heat, thereby delaying and reducing the increase in core temperature. METHODS: Eight healthy male volunteers anesthetized with propofol and isoflurane were studied. Volunteers were allowed to cool passively until core temperature reached 33 degrees C. On one randomly assigned day, the isoflurane concentration was reduced, to provoke thermoregulatory arteriovenous shunt vasoconstriction; on the other study day, a sufficient amount of isoflurane was administered to prevent vasoconstriction. On each day, forced-air warming was then applied for 2 h. Peripheral (arm and leg) tissue heat contents were determined from 19 intramuscular needle thermocouples, 10 skin temperatures, and "deep" foot temperature. Core (trunk and head) heat content was determined from core temperature, assuming a uniform compartmental distribution. Time-dependent changes in peripheral and core tissue heat contents were evaluated using linear regression. Differences between the vasoconstriction and vasodilation study days, and between the peripheral and core compartments, were evaluated using two-tailed, paired t tests. Data are presented as means +/ SD; P < 0.01 was considered statistically significant. RESULTS: Cutaneous heat transfer was similar during vasoconstriction and vasodilation. Forced-air warming increased peripheral tissue heat content comparably when the volunteers were vasodilated and vasoconstricted: 48 +/- 7 versus 53 +/- 10 kcal/h. Core compartment tissue heat content increased similarly when the volunteers were vasodilated and vasoconstricted: 51 +/- 8 versus 44 +/- 11 kcal/h. Combining the two study days, the increase in peripheral and core heat contents did not differ significantly: 51 +/- 8 versus 48 +/- 10 kcal/h, respectively. Core temperature increased at essentially the same rate when the volunteers remained vasodilated (1.3 degrees C/h) as when they were vasoconstricted (1.2 degrees C/h). CONCLUSIONS: The authors failed to confirm their hypothesis that thermoregulatory vasoconstriction decreases cutaneous transfer of applied heat and restricts peripheral-to-core flow of heat in anesthetized subjects. The reported difference between intraoperative and postoperative rewarming efficacy may result from nonthermoregulatory anesthetic-induced vasodilation. PMID- 8712444 TI - Epidural meperidine after cesarean section. A dose-response study. AB - BACKGROUND: Epidural meperidine is effective for postoperative analgesia, but the optimum dose has not been evaluated. METHODS: Five doses of epidural meperidine (12.5, 25, 50, 75, and 100 mg) given at the first request for analgesia after cesarean section were compared. Visual analog pain scores, duration of analgesia as defined by time to first patient-controlled epidural analgesia demand, plasma concentrations of meperidine, side effects, and subsequent 24-h consumption of meperidine were evaluated. RESULTS: All doses were effective, but patients took longer to become pain-free after 12.5 mg (median 30 min) compared with 25 mg (median 12 min, P = 0.038), and duration of analgesia was shorter after 12.5 mg (median 83 min) compared with 25 mg (median 165 min, P = 0.0005). Increasing dose to more than 25 mg did not improve onset or duration of analgesia. Plasma concentrations of meperidine were less than minimum effective analgesia concentration for all doses except 100 mg. There was more frequent nausea (P = 0.004) and dizziness (P = 0.0002) after 100 mg compared with smaller doses. CONCLUSIONS: Epidural meperidine provides effective postoperative analgesia, although of relatively short duration. A single dose of 25 mg is superior to 12.5 mg, but there is no benefit from increasing the dose to 50 mg or greater. PMID- 8712445 TI - Acute pain and central nervous system arousal do not restore impaired hypoxic ventilatory response during sevoflurane sedation. AB - BACKGROUND: To quantify the effects of acute pain on ventilatory control in the awake and sedated human volunteer, the acute hypoxic ventilatory response was studied in the absence and presence of noxious stimulation before and during 0.1 minimum alveolar concentration sevoflurane inhalation. METHODS: Step decreases in end-tidal partial pressure of oxygen from normoxia into hypoxia (approximately 50 mmHg) were performed in 11 healthy volunteers. Four acute hypoxic ventilatory responses were obtained per subject: one in the absence of pain and sevoflurane (C), one in the absence of sevoflurane with noxious stimulation in the form of a 1-Hz electrical current applied to the skin over the tibial bone (C + P), one in the absence of pain during the inhalation of 0.1 minimum alveolar concentration sevoflurane (S), and one during 0.1 minimum alveolar concentration sevoflurane with noxious stimulation (S + P). The end-tidal partial pressure of carbon dioxide was held constant at a value slightly greater than baseline (44 mmHg). To assess the central nervous system arousal state, the bispectral index of the electroencephalogram was monitored. Values are mean +/- SE. RESULTS: Pain caused an increase in prehypoxic baseline ventilation before and during sevoflurane inhalation: C = 13.7 +/- 0.9 l.min-1, C + P = 16.0 +/- 1.0 l.min-1 (P < 0.05 vs. C and S), S = 12.7 +/- 1.2 l.min-1, and S + P = 15.9 +/- 1.1 l.min-1 (P < 0.05 vs. C and S). Sevoflurane decreased the acute hypoxic ventilatory response in the absence and presence of noxious stimulation: C = 0.69 +/- 0.20 l.min-1 (% change in arterial hemoglobin-oxygen saturation derived from pulse oximetry [SpO2])-1, C + P = 0.64 +/- 0.13 l.min-1.%SpO2(-1), S = 0.48 +/- 0.15 l.min-1.%SpO2(-1) (P < 0.05 vs. C and C + P) and S + P = 0.46 +/- 0.21 l.min-1.%SpO2(-1) (P < 0.05 vs. C and C + P). The bispectral indexes were C = 96.2 +/ 0.7, C + P = 97.1 +/- 0.4, S = 86.3 +/- 1.3 (P < 0.05), and S + P = 95.0 +/- 1.0. CONCLUSIONS: The observation that acute pain caused an increase in baseline ventilation with no effect on the acute hypoxic ventilatory response indicates that acute pain interacted with ventilatory control without modifying the effect of low-dose sevoflurane on the peripheral chemoreflex loop. Acute pain increased the level of arousal significantly during sevoflurane inhalation but did not restore the approximately 30% depression of the acute hypoxic ventilatory response by sevoflurane. The central nervous system arousal state per se did not contribute to the impairment of the acute hypoxic ventilatory response by sevoflurane. PMID- 8712446 TI - Analgesic efficacy of low-dose ketamine. Somatosensory-evoked responses in relation to subjective pain ratings. AB - BACKGROUND: Low-dose ketamine has been shown to exert analgesic effects. Whether ketamine-induced pain relief may be quantitated by somatosensory evoked cerebral potentials has not been established. METHODS: Thirty healthy volunteers were assigned randomly to one of three groups. Subjects of group 1 (n = 10, control) were given saline as placebo. In groups 2 (n = 10) and 3 (n = 10), intravenous ketamine (0.25 mg. kg-1 and 0.50 mg. kg-1, respectively) was administered. The following variables were recorded at baseline and for 50 min after drug administration: electroencephalographic (EEG) data, somatosensory-evoked late cortical responses (SEP) elicited by intracutaneous stimulation of the fingertip (2-3 fold pain threshold), heart rate, mean arterial blood pressure, and end tidal PETCO2 via a tight-fitting mask. Electroencephalographic spectral power in selected frequency bands and frequency percentiles were calculated from the spontaneous EEG segment preceding each somatosensory stimulus. Somatosensory evoked late cortical response parameters were calculated from the respective poststimulus EEG segments. After recording of each EEG response, subjects were asked to rate the individual pain sensation. RESULTS: In group 1, all variables did not change over time. Ketamine administration resulted in dose-dependent decreases in alpha-activity and increases in theta power (group 2: 190%, group 3: 440%). Electroencephalographic changes were not related to changes in pain perception. For the first 30 min after ketamine injection, a dose-dependent decrease of the long-latency N150-P250 somatosensory-evoked late cortical response component was observed (group 2: 15-20%; group 3: 25-30%). Subjective pain ratings were also different between groups, with a higher degree of pain relief in group 3 for the first 30 min. At the end of the observation period, pain relief and the N150-P250 amplitude were comparable in both ketamine groups. CONCLUSIONS: These data indicate that pain relief induced by low-dose ketamine is dose-dependent for the first 30 min after bolus injection. Changes in pain perception may be quantitated by somatosensory-evoked cortical responses. Also, EEG changes are not specific for changes in nociception, but the increase in theta power may reflect the hypnotic effect of low-dose ketamine. PMID- 8712447 TI - Interaction between intrathecal neostigmine and epidural clonidine in human volunteers. AB - BACKGROUND: alpha 2-Adrenergic agonists are thought to produce analgesia, in part, by activating spinal acetylcholine release. The purpose of the current study was to examine the interaction between intrathecal neostigmine and epidural clonidine for analgesia and side effects in humans. METHODS: A total of 58 volunteers received an intrathecal injection of 5% dextrose in normal saline (D5NS) or neostigmine (50, 100, or 200 micrograms in D5NS), followed in 1 h by epidural saline or clonidine (computer-controlled infusion targeted to 50, 100, 200, or 400 ng/ml in cerebrospinal fluid) using an isobolographic design. Visual analog scale pain to a noxious cold stimulus, nausea, weakness, sedation, and other safety variables was measured before and at specified intervals after drug administration. RESULTS: The first 21 volunteers randomized to receive intrathecal hyperbaric neostigmine rather than D5NS received the drug while in the sitting position, and had none-to-minimal analgesia 1 h later. The remaining volunteers received the drug while in the lateral position, and demonstrated dose dependent analgesia in the foot 1 h later. Epidural clonidine also caused dose dependent analgesia. The combination of neostigmine and clonidine resulted in an additive enhancement for analgesia, but no enhancement of each drug's side effects, and a reduction in clonidine-induced hypotension. Neostigmine injected into subjects in the lateral position diminished clonidine-induced reductions in blood pressure and plasma norepinephrine. CONCLUSION: These results support enhancement of alpha 2-adrenergic analgesia by intrathecal neostigmine, but do not demonstrate synergy, as observed in animals. Lack of enhancement of side effects suggests this combination may be clinically useful. PMID- 8712448 TI - Density of lumbar cerebrospinal fluid in pregnant and nonpregnant humans. AB - BACKGROUND: Dextrose-free local anesthetics and opioids, alone and in combinations, are being used increasingly to provide subarachnoid anesthesia and analgesia. These dextrose-free drugs have been described as hypobaric by some and isobaric by others. To accurately classify anesthetics with regard to baricity, the density of cerebrospinal fluid (CSF) must be known. The authors sought to determine the exact density of human CSF, and determine whether CSF density is altered by pregnancy. METHODS: Density measurements accurate to 0.00001 g/ml were made at 37.00 degrees C, using a mechanical oscillation resonance frequency density meter. Cerebrospinal fluid samples were obtained from 44 patients during spinal anesthesia. Five groups were studied: men, and premenopausal, postmenopausal, term pregnant, and postpartum women. RESULTS: Mean CSF densities in men (1.00064 +/- 0.00012 g/ ml), postmenopausal women (1.00070 +/- 0.00018 g/ml), and nonpregnant premenopausal women (1.00049 +/- 0.00004 g/ ml) were significantly greater than in term pregnant (1.00030 +/- 0.00004 g/ml) and postpartum (1.00034 +/- 0.00005 g/ml) women. Cerebrospinal fluid density did not correlate with age. CONCLUSIONS: Mean CSF density varies in different patient subpopulations. Pregnancy and the immediate postpartum period are associated with the lowest CSF densities. In addition, the cutoff values defining hypobaricity (mean CSF density minus three standard deviations) are greater than previously reported. Accurate CSF density values should be used when considering baricity as a mechanism for clinical observations of dextrose-free intrathecal local anesthetics and opioids. Gestational status also should be considered. PMID- 8712449 TI - Sustained-release morphine for epidural analgesia in rats. AB - BACKGROUND: Epidural opioid analgesia often requires either continuous infusion or repeated injections, which are inconvenient for patients, increase risk of infection, and consume expensive physician and nursing time. In addition, potential respiratory depression is a major safety concern. The authors studied whether a single dose of epidurally administered, sustained-release morphine could prolong analgesia and reduce toxic effects in rats. METHODS: Sustained release morphine (DTC401) was prepared by encapsulating morphine sulfate in DepoFoam (Dep. Tech, San Diego, CA), a lipid-based, sustained-release drug delivery system. A standard hot-plate test for analgesia, pulse oximetry for hemoglobin oxygen saturation, corneal-reflex loss, and incidence of catalepsy were used to assess efficacy and toxicities. Cerebrospinal fluid and serum pharmacokinetic studies were performed after a single epidural dose, using a commercially available radioimmunoassay kit. RESULTS: Single epidural doses of DTC401 resulted in equivalent onset time to peak analgesia but significantly prolonged analgesia compared with morphine sulfate. Hemoglobin oxygen saturation was decreased minimally, and the incidences of catalepsy and corneal-reflex loss were minimal, even at large doses of DTC401. In contrast, the larger doses of morphine sulfate significantly decreased hemoglobin oxygen saturation, and caused catalepsy and loss of the corneal-reflex. The Cmax for DTC401 was 32% in cerebrospinal fluid and 6% in serum, relative to morphine sulfate. The terminal half-life for DTC401 was increased 32 fold in the cerebrospinal fluid compared with morphine sulfate. CONCLUSIONS: A single epidural dose of DTC401, compared with morphine sulfate, prolonged duration of analgesia, with minimal supraspinal toxic effects, in rats. PMID- 8712450 TI - Heparinase I (neutralase) reversal of systemic anticoagulation. AB - BACKGROUND: Protamine causes multiple adverse reactions. Heparinase I, a specific enzyme that inactivates heparin, is a possible alternative to protamine. In this study, the authors examined the efficacy of heparinase I to reverse heparin induced anticoagulation in vitro and compared heparinase I to protamine as an antagonist of heparin-induced anticoagulation in dogs. METHODS: In the in vitro study, blood was obtained from the extracorporeal circuits of 12 patients, and activated clotting times were determined after adding different concentrations of heparinase I. In the in vivo study, 24 anesthetized dogs received 300 units/kg heparin injected intravenously for 5 s, then 10 min later, 3.9 mg/kg protamine, 5 41 micrograms/kg heparinase I, or the vehicle (n = 4/group) were administered intravenously, and activated clotting times and hemodynamics were measured. RESULTS: In the in vitro study, heparin concentrations of 3.3 +/- 1.0 (mean +/- SD) units/ml (approximately 0.033 mg/ml; n = 12) were reversed in the blood of patients by heparinase I at concentrations > 0.490 microgram/ml. In the canine study, heparinase at all doses studied and protamine effectively reversed the anticoagulating effects of heparin within 10 min of administration. Protamine produced adverse hemodynamic effects, whereas heparinase or its vehicle produced no significant change in arterial pressure. CONCLUSION: Both heparinase I and protamine effectively reversed heparin anticoagulation. However, as opposed to protamine, heparinase I did not produce any significant hemodynamic changes when given as a bolus to dogs. PMID- 8712451 TI - Effects of halothane and isoflurane on carbon monoxide-induced relaxations in the rat aorta. AB - BACKGROUND: Halothane and isoflurane previously were reported to attenuate endothelium-derived relaxing factor/nitric oxide-mediated vasodilation and cyclic guanosine monophosphate (cGMP) formation in isolated rat aortic rings. Carbon monoxide has many chemical and physiologic similarities to nitric oxide. This study was designed to investigate the effects of halothane and isoflurane on carbon monoxide-induced relaxations and cGMP formation in the isolated rat aorta. METHODS: Isometric tension was recorded continuously from endothelium denuded rat aortic rings suspended in Krebs-filled organ baths. Rings precontracted with submaximal concentrations of norepinephrine were exposed to cumulative concentrations of carbon monoxide (26-176 microM). This procedure was repeated three times, with anesthetics delivered 10 min before the second procedure. Carbon monoxide responses of rings contracted with the same concentration of norepinephrine (10(-6) M and 2 x 10(-6) M) used in the anesthetic-exposed preparations also were examined. The concentrations of cGMP were determined in denuded rings using radioimmunoassay. The rings were treated with carbon monoxide (176 microM, 30 s) alone, or carbon monoxide after a 10-min incubation with halothane (0.34 mM or 0.72 mM). To determine whether the sequence of anesthetic delivery influenced results, vascular rings pretreated with halothane were compared with nonpretreated rings. RESULTS: Carbon monoxide (26-176 microM) caused a dose-dependent reduction of norepinephrine-induced tension, with a maximal relaxation of 1.51 +/- 0.07 g (85 +/- 7% of norepinephrine-induced contraction). Halothane (0.34 mM and 0.72 mM) significantly attenuated the carbon monoxide-induced relaxations, but only the highest concentration of isoflurane (0.53 mM) significantly attenuated the carbon monoxide-induced relaxations. Carbon monoxide (176 microM) significantly increased cGMP content (+88.1 +/- 7.1%) and preincubation of the aortic rings with halothane (0.34 mM and 0.72 mM) inhibited this increase (-70.7 +/- 6.8% and -108.1 +/- 10.6%, respectively). When aortic rings and carbon monoxide were added simultaneously to Krebs solution equilibrated with halothane (0.72 mM), no inhibition of cGMP formation occurred. CONCLUSION: Carbon monoxide-induced endothelium-independent relaxations of rat aortic rings were decreased by clinically relevant concentrations of halothane and isoflurane. The carbon monoxide-induced elevations of cGMP were attenuated by halothane only when the anesthetic was incubated with aortic rings before carbon monoxide treatment. The possible clinical significance of the actions of the anesthetics on this endogenous vasodilator is yet to be determined. PMID- 8712452 TI - Equianalgesic doses of subcutaneous but not intrathecal morphine alter phenotypic expression of cell surface markers and mitogen-induced proliferation in rat lymphocytes. AB - BACKGROUND: Surgical trauma and opioids are linked with suppression of immune function. Evidence suggests a probable supraspinal action of morphine in altering immune function, although the role of spinal systems have not been evaluated. Therefore, this study compared the effect of equianalgesic doses of subcutaneous and intrathecal morphine on lymphocyte proliferative responses and phenotypic expression of lymphocyte cell surface markers in rats. METHODS: Equianalgesic doses of subcutaneous (10 mg/kg) or intrathecal (30 micrograms, by a chronic intrathecal catheter) morphine were given twice for 5 h (time 0 and 2.5 h). Immediately after the 5-h period or 24 h after the initial injection, spleens were harvested and lymphocytes isolated. Mitogen-induced (phytohemagglutinin, concanavalin A, pokeweed, lipopolysaccharide) lymphocyte proliferation and monoclonal antibody labeling of cell surface markers (T cell, B cell, CD4+, CD8+) were then performed. RESULTS: Subcutaneous morphine acutely suppressed lymphocyte proliferation to the mitogens phytohemagglutinin, pokeweed, and concanavalin A by 37%, 21%, and 20% respectively; however, proliferative responses returned to baseline within 24 h. Morphine treatment did not alter the response to lipopolysaccharide. The number of splenic lymphocytes also decreased, whereas the percentage of lymphocytes expressing the CD4+ marker (T helper/inducer cells) modestly increased. Intrathecal morphine did not alter lymphocyte proliferative responses, nor did it change phenotypic expression of cell surface markers. CONCLUSIONS: Subcutaneous morphine inhibited lymphocyte proliferation, decreased splenic lymphocyte number, and altered phenotypic expression of cell surface markers, whereas equianalgesic doses of intrathecal morphine did not. Although these results suggest that spinal opioids may have theoretical benefits for the analgesic management of immunocompromised patients, further studies are clearly indicated. PMID- 8712453 TI - Effects of halothane and isoflurane on bradykinin-evoked Ca2+ influx inbovine aortic endothelial cells. AB - BACKGROUND: Volatile anesthetics, such as halothane and isoflurane, have been reported to affect the endothelium mediated relaxation of vascular smooth muscle cells. Because the activity of the constitutive nitric oxide synthase in endothelial cells depends on the availability of intracellular Ca2+, there is a definite possibility that the observed inhibitory effect of volatile anesthetics involves an action on the agonist-evoked internal Ca2+ mobilization and/or Ca2+ influx in these cells. Therefore, a study was undertaken to determine how halothane and isoflurane affect the Ca2+ signalling process in vascular endothelial cells. METHODS: The effect of halothane and isoflurane on the Ca2+ response to bradykinin of bovine aortic endothelial (BAE) cells was investigated using the fluorescent Ca2+ indicator fura-2. Halothane or isoflurane was applied either to resting cells or after bradykinin stimulation. The agonist-evoked Ca2+ influx in BAE cells was estimated by measuring either the rate of fura-2 quenching induced by Mn2+ or the increase in cytosolic Ca2+ concentration initiated after readmission of external Ca2+ after a brief exposure of the cells to a Ca(2+)-free external medium. The effects of halothane on cell potential and intracellular Ca2+ concentration were measured in cell-attached patch-clamp experiments in which a calcium-activated K+ channel and an inward rectifying Ca(2+)-independent K+ channel were used as probes to simultaneously monitor the intracellular Ca2+ concentration and the cell transmembrane potential. In addition, combined fura-2 and patch-clamp cell-attached recordings were carried out, to correlate the variations in internal Ca2+ caused by halothane and the activity of the Ca(2+)-dependent K+ channels, which are known in BAE cells to regulate intracellular potential. Finally, a direct action of halothane and isoflurane on the gating properties of the Ca(2+)-activated K+ channel present in these cells was investigated in patch-excised inside-out experiments. RESULTS: The results of the current study indicate that the initial Ca2+ increase in response to bradykinin stimulation is not affected by halothane, but that pulse applications of halothane (0.4-2 mM) or isoflurane (0.5-1 mM) reversibly reduce the sustained cytosolic Ca2+ increase initiated either by bradykinin or by the Ca2+ pump inhibitor thapsigargin. In addition, halothane appeared to dose dependently inhibit the Ca2+ influx evoked by bradykinin, and to cause, concomitant to a decrease in cytosolic Ca2+ concentration, a depolarization of the cell potential. Halothane failed, however, to affect internal Ca2+ concentration in thapsigargin-treated endothelial cells, which were depolarized using a high K+ external solution. Finally, halothane and isoflurane decreased the open probability of the Ca(2+)-dependent K+ channel present in these cells. CONCLUSIONS: These observations suggest that the effects of halothane and isoflurane on Ca2+ homeostasis in BAE cells reflect, for the most part, a reduction of the thapsigargin- or bradykinin-evoked Ca2+ influx, which would be consequent to a cellular depolarization caused by an inhibition of the Ca(2+) dependent K+ channel activity initiated after cell stimulation. PMID- 8712454 TI - Phenylephrine increases cerebral blood flow during low-flow hypothermic cardiopulmonary bypass in baboons. AB - BACKGROUND: Although low-flow cardiopulmonary bypass (CPB) has become a preferred technique for the surgical repair of complex cardiac lesions in children, the relative hypotension and decrease in cerebral blood flow (CBF) associated with low flow may contribute to the occurrence of postoperative neurologic injury. Therefore, it was determined whether phenylephrine administered to increase arterial blood pressure during low-flow CPB increases CBF. METHODS: Cardiopulmonary bypass was initiated in seven baboons during fentanyl, midazolam, and isoflurane anesthesia. Animals were cooled at a pump flow rate of 2.5 l.min 1.m-2 until esophageal temperature decreased to 20 degrees C. Cardiopulmonary bypass flow was then reduced to 0.5 l.min-1.m-2 (low flow). During low-flow CPB, arterial partial pressure of carbon dioxide (Pco2) and blood pressure were varied in random sequence to three conditions: (1) Pco2 30-39 mmHg (uncorrected for temperature), control blood pressure; (2) Pco2 50-60 mmHg, control blood pressure; and (3) Pco2 30-39 mmHg, blood pressure raised to twice control by phenylephrine infusion. Thereafter, CPB flow was increased to 2.5 l.min-1.m-2, and baboons were rewarmed to normal temperature. Cerebral blood flow was measured by washout of intraarterial 133Xe before and during CPB. RESULTS: Phenylephrine administered to increase mean blood pressure from 23 +/- 3 to 46 +/- 3 mmHg during low-flow CPB increased CBF from 14 +/- 3 to 31 +/- 9 ml.min-1.100 g-1, P < 0.05. Changes in arterial Pco2 alone during low flow bypass produced no changes in CBF. CONCLUSIONS: Although low-flow CPB resulted in a marked decrease in CBF compared with prebypass and full-flow bypass, phenylephrine administered to double arterial pressure during low-flow bypass produced a proportional increase in CBF. PMID- 8712455 TI - Genetic differences affecting the potency of stereoisomers of isoflurane. AB - BACKGROUND: In previous studies, researchers demonstrated the ability of a variety of organisms and in vitro sites of anesthetic action to distinguish between stereoisomers of isoflurane or halothane. However, it was not shown whether organisms with differing sensitivities to stereoisomers of one volatile anesthetic are able to distinguish between stereoisomers of another. In this study, the responses of mutants of Caenorbabditis elegans to stereoisomers of isoflurane were determined for comparison to previous results in halothane. METHODS: Mutant strains of C. elegans were isolated and grown by standard techniques. The EC50s (the effective concentrations of anesthetia at which 50% of the animals are immobilized for 10 s) of stereoisomers of isoflurane and the racemate were determined in wild type and mutant strains of C. elegans. RESULTS: Wild type C. elegans and strains with high EC50S of the racemate were more sensitive to the (+) isomer of isoflurane by approximately 30%. The racemate showed a EC50s similar to the less potent isomer, the (-) form. In the strains with low EC50s, one strain showed no ability to differentiate between the stereoisomers, whereas two showed a 60% difference between the (+) and (-) forms. CONCLUSIONS: The ability to distinguish between stereoisomers of isoflurane is associated with genetic loci separate from those that distinguish between stereoisomers of halothane. These results are consistent with multiple sites of action for these anesthetics. PMID- 8712456 TI - Effects of intravenous anesthetics on atrial wavelength and atrioventricular nodal conduction in guinea pig heart. Potential antidysrhythmic properties and clinical implications. AB - BACKGROUND: Supraventricular tachydysrhythmias such as atrial fibrillation frequently complicate the perioperative period. Two electrophysiologic factors critical to the pathogenesis of supraventricular tachydysrhythmias are: 1) atrial wavelength, the product of atrial conduction velocity (CV) and effective refractory period (ERP), and 2) atrioventricular nodal conduction. Modulation of these factors by drugs has important clinical ramifications. The authors studied the effects of propofol, thiopental, and ketamine on atrial wavelength and atrioventricular nodal function in guinea pig isolated atrial trabeculae and hearts, respectively. METHODS: Electrocardiogram recordings in superfused atrial tissue were obtained using hanging microelectrodes. A stimulating and two recording electrodes were placed on a single atrial trabecula, and the interelectrode distance was measured. Atrial ERP determinations were made using a premature stimulus protocol. The time (t) required for a propagated impulse to traverse the interelectrode distance (d) was measured. Conduction velocity was calculated as d/t. Langendorff-perfused guinea pig hearts were instrumented for low atrial pacing (cycle length = 300 ms) and for measurements of stimulusto-His bundle interval, an index of atrioventricular nodal conduction. To investigate the frequency-dependent behavior of the atrioventricular node, computer-based measurements were made of Wenckebach cycle length (WCL) and atrioventricular nodal ERP. RESULTS: Thiopental significantly prolonged atrial ERP in a concentration-dependent manner, whereas propofol and ketamine had no significant effect on atrial refractoriness. In contrast, ketamine caused a dose-dependent decrease in atrial CV, but propofol and thiopental had no significant effect on CV. Therefore, thiopental, ketamine, and propofol caused an increase, a decrease, and no change, respectively, in atrial wavelength. All anesthetics caused a concentration-dependent prolongation of the stimulus-to-His bundle interval, atrioventricular nodal ERP, and WCL. However, on an equimolar basis, significant differences in potencies were found. The concentrations of drug that caused a 20% increase in ERP (ERP20) and WCL (WCL20) for propofol, thiopental, and ketamine were 14 +/- 2 microM, 26 +/- 3 microM, and 62 +/- 11 microM, and 17 +/- 2 microM, 50 +/- 1 microM, and 123 +/- 19 microM (mean +/- SEM), respectively. Therefore, the rank order of potency for frequency-dependent atrioventricular nodal effects is propofol > thiopental > ketamine. CONCLUSION: The authors' results indicate that propofol would be most effective at filtering atrial impulses during supraventricular tachydysrhythmias, whereas thiopental would be most effective at preventing atrial reentrant dysrhythmias. In contrast, ketamine may be most likely to promote atrial reentry while having minimal effect on atrioventricular nodal conduction. PMID- 8712458 TI - Iconography in anesthesiology. The importance of society seals in the 1920s and 30s. PMID- 8712457 TI - Desflurane, sevoflurane, and isoflurane impair canine left ventricular-arterial coupling and mechanical efficiency. AB - BACKGROUND: The effects of desflurane, sevoflurane, and isoflurane on left ventricular-arterial coupling and mechanical efficiency were examined and compared in acutely instrumented dogs. METHODS: Twenty-four open-chest, barbiturate-anesthetized dogs were instrumented for measurement of aortic and left ventricular (LV) pressure (micromanometer-tipped catheter), dP/dtmax, and LV volume (conductance catheter). Myocardial contractility was assessed with the end systolic pressure-volume relation (Ees) and preload recruitable stroke work (Msw) generated from a series of LV pressure-volume diagrams. Left ventricular-arterial coupling and mechanical efficiency were determined by the ratio of Ees to effective arterial elastance (Ea; the ratio of end-systolic arterial pressure to stroke volume) and the ratio of stroke work (SW) to pressure-volume area (PVA), respectively. RESULTS: Desflurane, sevoflurane, and isoflurane reduced heart rate, mean arterial pressure, and left ventricular systolic pressure. All three anesthetics caused similar decreases in myocardial contractility and left ventricular afterload, as indicated by reductions in Ees, Msw, and dP/dtmax and Ea, respectively. Despite causing simultaneous declines in Ees and Ea, desflurane decreased Ees/Ea (1.02 +/- 0.16 during control to 0.62 +/- 0.14 at 1.2 minimum alveolar concentration) and SW/PVA (0.51 +/- 0.04 during control to 0.43 +/- 0.05 at 1.2 minimum alveolar concentration). Similar results were observed with sevoflurane and isoflurane. CONCLUSIONS: The present findings indicate that volatile anesthetics preserve optimum left ventricular-arterial coupling and efficiency at low anesthetic concentrations (< 0.9 minimum alveolar concentration); however, mechanical matching of energy transfer from the left ventricle to the arterial circulation degenerates at higher end-tidal concentrations. These detrimental alterations in left ventricular-arterial coupling produced by desflurane, sevoflurane, and isoflurane contribute to reductions in overall cardiac performance observed with these agents in vivo. PMID- 8712459 TI - Abdominal aortic compression to treat circulatory collapse caused by severe pulmonary hypertension during liver transplantation. PMID- 8712460 TI - Pneumocephalus after spinal anesthesia. PMID- 8712461 TI - Gastroesophageal perforation after intraoperative transesophageal echocardiography. PMID- 8712462 TI - Spinal cord infarction after surgery in a patient in the hyperlordotic position. PMID- 8712464 TI - Enoxaparin and epidural analgesia. PMID- 8712463 TI - Nonanesthetic haloalkanes and nicotinic acetylcholine receptor desensitization kinetics. PMID- 8712465 TI - Intraoperative disarticulation of a triple-lumen central catheter. PMID- 8712467 TI - Teaching airway management skills: what about patient consent? PMID- 8712466 TI - After transdermal fentanyl: acute toxic delirium or central anticholinergic syndrome? PMID- 8712468 TI - Tracheal intubation through the laryngeal mask airway. PMID- 8712469 TI - Use of an 8-F catheter to assist with Bullard laryngoscopy in intubating the trachea. PMID- 8712470 TI - Practice guidelines and treatment of patients with von Willebrand's disease. PMID- 8712471 TI - In defense of Waters, Blalock, and Taylor. PMID- 8712472 TI - Venous gas embolism from an Argon coagulator. PMID- 8712473 TI - A useful maneuver when intravenous access is difficult. PMID- 8712474 TI - Infectious complications after epidural anesthesia. PMID- 8712475 TI - Color of compressed gas cylinders. PMID- 8712476 TI - Clarification of kinetic terminology. PMID- 8712477 TI - Acute myocardial infarction during pregnancy and puerperium: a review. AB - The purpose of this review is to analyze the possible parameters that lead to the development of what is a rare event--acute myocardial infarction (AMI) during pregnancy and puerperium. Through the Index Medicus, 109 publications on the subject were obtained. Since the first well-documented case by Katz in 1922, 136 patients have been reported, and from these reports the following data have been gathered: the average age was 32.1 years. This event is more frequent during the third trimester and puerperium of the first and second pregnancies. In 42.6% of the patients no coronary risk factors were observed, but when present, hypertension and cigarette smoking were the most common. The anterior wall along or in combination with any other anatomic area was affected in 73% of cases. Coronary angiograms, when taken, appeared normal in 47%. The maternal mortality rate was 26/136 (19.1%) and was higher during the third trimester, labor, and puerperium. Eight patients (8/26) (30.7%) had sudden death. In 5 of these, (62.5%) coronary thrombosis was found. In 18/26 deaths, an autopsy was performed; 9/18 (50%) had coronary thrombus formation and in 7/18 (39%) variable degrees of atherosclerosis were detected. On the other hand, the fetal mortality rate was 16.9%; however, in only 52% was death coincidental with that of the mother. Coronary artery spasm associated with a probable hypercoagulability state was the most likely mechanism in the majority of these patients, followed by atherosclerotic heart disease and coronary dissection-the last being secondary most likely to hormonal changes. During the AMI these patients should be studied by a medical team composed of a cardiologist, gynecologist, and anesthesiologist. A complete cardiologic work-up should be made to decide individually about further pregnancies. PMID- 8712478 TI - Leukocyte flow properties, polymorphonuclear membrane fluidity, and cytosolic Ca2+ content in subjects with vascular atherosclerotic disease with and without noninsulin-dependent diabetes mellitus. AB - The aim of this research was the evaluation of white blood cell (WBC) filtration, reflecting WBC flow properties, polymorphonuclear cell membrane fluidity, and cytosolic Ca2+ content in subjects with vascular atherosclerotic disease (VAD) and in VAD subjects with noninsulin-dependent diabetes mellitus (NIDDM), in good hemodynamic balance. The authors examined WBC filtration (unfractionated, mononuclear [MN], polymorphonuclear [PMN] cells), using the St. George Filtrometer and considering, respectively, the initial relative flow rate (IRFR) and the clogging rate (CR); the PMN membrane fluidity, employing the fluorescent probe TMA-DPH and calculating the fluorescence polarization degree; and the PMN cytosolic Ca2+ content, adopting the fluorescent probe Fura 2-AM and considering the ratio between the Fura 2-Ca2+ complex and the unchelated Fura 2 fluorescence intensity. The obtained data showed that only the filtration parameters (IRFR, CR) of unfractionated WBCs discriminated normal subjects from VAD groups, whereas the filtration parameters of MN and PMN cells did not demonstrate any distinction. PMN membrane fluidity did not distinguish normal subjects from VAD groups, whereas PMN cytosolic Ca2+ content was significantly increased in VAD groups in comparison with normal subjects. No relationship was evident between WBC filtration and plasma metabolic parameters; the correlations obtained between PMN filtration and other PMN parameters need further investigation. PMID- 8712479 TI - Persisting effect of Ca(2+)-channel blockers on left ventricular function in hypertrophic cardiomyopathy after 14 years' treatment. AB - Ca(2+)-channel blockers of the verapamil type have been reported to exert a beneficial effect on clinical symptoms and survival rates in hypertrophic cardiomyopathy. The effects of verapamil have been attributed predominantly to an improved diastolic filling. It is unknown whether an effect on diastolic filling persists in these patients after long-term treatment. Fourteen patients (12 men, 2 women, median age fifty-one [thirty-two to fifty-five] years) with hypertrophic cardiomyopathy were included in the study. Patients had been treated with verapamil 240-480 mg/d or gallopamil 150-200 mg/d for fourteen (seven to seventeen) years. The effect of a withdrawal of Ca(2+)-channel blockers on parameters of left ventricular diastolic function was evaluated at rest and during exercise in patients with hypertrophic cardiomyopathy after long-term therapy. Investigations were performed at rest and during supine ergometric exercise during ongoing Ca(2+)-channel blocking therapy and after five (four to nine) days' withdrawal (control). Pulsed Doppler echocardiography was used to record diastolic mitral flow profiles from an apical four-chamber view. Withdrawal of Ca(2+)-channel blockers of the phenylalkylamine type after long term treatment of hypertrophic cardiomyopathy resulted in a significant reduction of early diastolic inflow velocity at rest and during exercise. In conclusion, these results indicate a persistent improvement of early diastolic filling by Ca(2+)-channel blockers even after long-term treatment. PMID- 8712480 TI - Compensatory enlargement of angiographically normal coronary segments in patients with coronary artery disease. In vivo documentation using intravascular ultrasound. AB - Intravascular ultrasound (IVUS) frequently reveals plaque formation at sites with a normal angiographic appearance. However, whether angiographically normal coronary arteries undergo adaptive expansion in vivo remains uncertain. The authors studied 12 patients (11 men, 1 woman; mean age fifty-three +/- ten years [mean +/- SD]) with focal coronary stenosis. Sixty IVUS images from angiographically normal coronary segments were analyzed (14 left main, 44 left anterior descending, and 2 left circumflex coronary arteries). The mean percent area stenosis was 36 +/- 5% and the circular shape factor of the lumen cross section averaged 0.97 +/- 0.02. Both total arterial area and internal elastic lamina area increased as the plaque area expanded (y = 2.13x + 8.07, r = 0.87, P = 0.0001; y = 2.06x + 4.57, r = 0.87, P = 0.0001, respectively), suggesting that for every 1 mm2 increase in plaque area, the total arterial area increased by approximately 2.13 mm2 and the internal elastic lamina area increased by approximately 2.06 mm2. The lumen area also increased as the plaque area expanded (y = 1.06x + 4.57, r = 0.68, P = 0.0001), suggesting that for every 1 mm2 increase in plaque area, the lumen area increased by approximately 1.06 mm2. The medial area did not correlate with the plaque area (r = 0.15, P = 0.26). Thus, compensatory enlargement precedes development of angiographically, detectable coronary atherosclerosis. Furthermore, in early stages of atherosclerosis, arterial enlargement may overcompensate for plaque area. The reduction of the total medial mass does not appear to contribute to the mechanism of compensatory enlargement. PMID- 8712481 TI - Do physicians' beliefs about coronary risk factors account for poor screening and treatment rates? AB - The authors hypothesized that the widely observed low rates of coronary risk factor screening and treatment among cardiologists could be due to lack of belief in the importance of risk factors. They performed an audit of 160 randomly selected medical records of patients with coronary artery disease and surveyed the attending physicians' ranking of importance of individual risk factors to determine the extent of risk factor screening and treatment done by these physicians. Screening and treatment rates for individual risk factors were highly variable but generally well below expected. There was no correlation among rank orders of screening rates, treatment rates, and physician-rated importance of each risk factor. These results suggest that lack of belief in risk factor importance does not explain failure of physicians to screen for and treat risk factors in coronary patients. PMID- 8712482 TI - Wine consumption and other dietary variables in males under 60 before and after acute myocardial infarction. AB - In France there are few cardiac deaths in spite of high animal fat intake. France and Italy have the highest overall intake of alcohol in the world. Obviously, there is an inverse association between coronary heart disease (CHD) and alcohol intake in these countries. Although in the past decade several-large scale population studies have confirmed the beneficial effect of alcohol on CHD, these studies may not have been sensitive to control all the confounding variables. No one so far has explored the possibility that the French may be protected by their low level of life stress. In 1993 we conducted a case-control study (n = 118) to examine psychological variables in a group of Dutch males under sixty years of age, before and after acute myocardial infarction (MI). After adjustment for total cholesterol, blood pressure, and smoking, a number of psychological factors appeared to be independently associated with an increased risk of MI. For the present study the same group of patients was assessed for consumption of different types of alcoholic beverages, coffee, sugar, high-fat diet, and vegetables. In the univariate analysis patients appeared to have consumed more red wine (odds ratio [OR] 0.2, P = 0.03) and controls more spirits (OR 4.0, P = 0.005). After adjustment for total cholesterol, blood pressure, and smoking as well as the independent psychological factors, red wine lost its significance (OR 0.4, P = 0.17) whereas the OR for spirits even rose (OR 6.0, P = 0.01). The beneficial effect of wine may be an expression of a relatively low level of life stress. Alcohol itself is not protective but rather a strong risk factor of MI. PMID- 8712483 TI - Left atrial size changes in patients with paroxysmal lone atrial fibrillation. An echocardiographic follow-up. AB - Left atrial enlargement has been demonstrated to occur as a consequence of the arrhythmia in patients with chronic atrial fibrillation (AF) in the absence of organic heart disease, whereas contrasting results have been reported in patients with paroxysmal lone AF. In the present investigation the behavior of left atrial size was followed up for an average period of 30.3 months in 20 patients with paroxysmal lone AF since their first arrhythmic episode. No significant changes in left atrial size were found at the end of the follow-up period. In 65% of patients the arrhythmia recurred at least once yearly. Left atrial size at enrollment was not significantly different in these patients from those without recurrences and did not change in either group during follow-up. The results suggest that in patients with lone AF left atrial dilatation occurs only after the arrhythmia becomes chronic. Early restoration of sinus rhythm may interrupt the vicious circle leading to atrial enlargement. PMID- 8712484 TI - Cortical blindness following coronary arteriography: a rare but self-cured complication. Two case reports. AB - The rare and self-cured complication of cortical blindness following coronary arteriography is presented in 2 patients who underwent cardiac catheterization. Both patients were submitted to an aortocoronary bypass grafting procedure a few years before and were under clinical investigation for a new onset of unstable angina pectoris. PMID- 8712485 TI - A massive calcified left ventricular aneurysm with normal coronary arteries and without myocardial infarction. A case report. AB - Most cardiac aneurysms develop after myocardial infarction. Calcification in the aneurysmal wall is seen rarely. In this case report the authors present a thirty nine-year-old man, who had been free from symptoms until eight months before, when he began to experience palpitations due to monomorphic sustained ventricular tachycardia. A chest roentgenogram disclosed an oval calcification on the left ventricular apex. Coronary angiography and left ventriculography revealed normal epicardial coronary arteries and a massive calcified and ossified left ventricular apical aneurysm. He had no chest pain, nor were there electrocardiographic findings of myocardial infarction. Attacks of ventricular tachycardia disappeared after aneurysmectomy was performed. To the author's knowledge there is no case report in the literature of a calcified left ventricular aneurysm with normal epicardial coronary arteries and without clinical and electrocardiographic findings of infarction. They discuss the possible etiology of this case. PMID- 8712486 TI - Acute myocardial infarction with normal coronary arteries in a patient with dermatomyositis. Case report. AB - The authors present a forty-year-old man, with a history of dermatomyositis for the past twelve years, managed intermittently with prednisone. During an exacerbation of his illness he developed an acute anterior wall myocardial infarction with normal coronary arteries. A literature review indicates this represents a rare association. PMID- 8712487 TI - Myocardial infarction in myxoma patients with normal coronary arteries. Case reports. AB - Three cases of patients with cardiac myxoma who had attacks of acute myocardial infarction are presented. Cineangiographic study showed normal coronary arteries. Immunohistochemical and serologic examination revealed that both interleukin-6 and interleukin-8 were secreted in cardiac myxoma. The authors discuss the relation between these cytokines and myocardial infarction with normal coronary arteries. PMID- 8712488 TI - Significance of dual left anterior descending coronary artery in interventional cardiology. A case report. AB - The variants of the left anterior descending (LAD) coronary artery are important in interpretation of coronary angiograms and in interventional procedures. The authors present a patient who had percutaneous transluminal coronary angioplasty and stent implantation to a presumed proximal LAD without the realization that dual LAd was present because of the total occlusion of the long LAD. PMID- 8712489 TI - Partitioning the components of maxillary tooth displacement by the comparison of data from three cephalometric superimpositions. AB - Using roentgenographic cephalograms from a sample of subjects with metallic implants, appropriately superimposed tracings were used to distinguish developmental and treatment-associated displacements of the maxillary central incisor and first molar associated "local" changes within the periodontium from "secondary" changes which reflect sutural and appositional growth at more distant osseous loci. Tracings were superimposed on anterior cranial base (ACB), on the maxillary implants only (IMP_MAX), and according to the best fit of maxillary anatomic structures without reference to the implants (A_MAX). Using the IMP_MAX superimposition, one could measure total local displacement at any landmark taking into consideration the effects of all appositional and resorptive changes on the superior and anterior surfaces of the palate, whereas using the A_MAX superimposition one could measure local displacement without consideration of surface appositional and resorptive changes. If the second of these measurements were subtracted from the first, the result would be a direct measurement of the effects of surface appositional and resorptive changes as they are expressed at that particular landmark. This strategy has enabled us to quantify and report the amount of accommodation which occurs at the location of each dental landmark in association with the resorptive and appositional changes which occur through time on the superior and anterior surfaces of the hard palate. PMID- 8712490 TI - Association between the direction of orthopedic headgear force and sutural responses in the nasomaxillary complex. AB - This study was designed to investigate biomechanical responses of the sutures in the nasomaxillary complex to orthopedic headgear forces applied in various directions. A three-dimensional analytic model of the craniofacial complex was used for finite element analysis. A posteriorly-directed force of 1.0 Kgf was applied to the maxillary first molars in 30 degrees inferior, parallel, and 30 degrees, 52.4 degrees and 60 degrees superior directions to the functional occlusal plane. Mean principal and shear stresses were evaluated at the sphenozygomatic, temporozygomatic, sphenomaxillary, frontomaxillary and frontozygomatic sutures and lamina cribrosa. As the force direction passed closer to the center of resistance (CRe) of the complex (52.4 degrees superior direction). normal stresses approached a certain level of uniform compressive stress (-2.5 gf/mm2) with gradual decrease in shear stresses, whereas variation in these stresses produced by the forces applied in other horizontal and inferior directions was greater. It is shown that stresses in the nasomaxillary sutures are varied by the direction of headgear force. Directing the line of force closer the CRe may produce the most optimal sutural modification effective for controlling forward and downward maxillary growth. PMID- 8712491 TI - A postretention study of patients presenting with a maxillary median diastema. AB - The purpose of this study was to assess postretention stability of maxillary midline diastema closure, to search for predictors of relapse, and to test for associations between relapse and other postretention changes. The sample consisted of 35 patients with pretreatment diastemas ranging from 0.9 mm to 3.0 mm (mean 1.4, SD = 0.5) following eruption of the maxillary canines. Data were gathered from treatment charts, study models, periapical radiographs, and cephalograms taken pretreatment, posttreatment, and 1 to 26 years postretention (mean 11.4, SD = 6.4). Measurable diastema relapse was observed in only 12 cases. The majority of the relapse was 0.6 mm or less, and maximum relapse was 3.0 mm. Abnormal frenums and/or intermaxillary osseous clefts did not appear to be risk factors for relapse, and no pretreatment predictors of relapse could be established. The only posttreatment change associated with diastema relapse was proclination of the maxillary incisors (p < 0.01). PMID- 8712492 TI - Stability of changes associated with chin cup treatment. AB - Twenty-four Japanese girls with anterior crossbite (Class III malocclusion) were selected for this study of the stability of changes associated with chin cup therapy. Pretreatment cephalometric measurements of the study sample were compared with those in a normal group. In addition, angular and linear measurements were also compared to assess the effectiveness of chin cup therapy in improving Class III skeletal components during the postretention period. The subjects showed characteristic values found in Class III malocclusion for SNB, ANB, and NPg to FH, and these values were significantly different from those in the normal group. The subjects who were past puberty showed more severe Class III skeletal patterns for ANB compared with the prepubertal subjects, and their initial Class III skeletal components showed more satisfactory improvement, including 2.0 degrees increase of SNA and 1.8 degrees increase of ANB during the postretention period. PMID- 8712493 TI - Transposition of teeth and genetic etiology. AB - Twenty-one cases of transposition are presented showing a crude prevalence of 0.4%. A high rate of bilateral occurrence was seen. A significant number of cases were associated with other dental anomalies, such as peg-shaped lateral incisors and overretained deciduous teeth. Genetic etiology has been stressed and various other theories discussed. PMID- 8712494 TI - Centric relation treatment and articulator mountings in orthodontics. PMID- 8712495 TI - Tooth numbering progress. PMID- 8712496 TI - Not all bone loss created equal. PMID- 8712497 TI - Skeletal Class III relationship. PMID- 8712498 TI - Case report SB: long-term follow-up on Class II treatment with first molar extractions. AB - Widespread acceptance of water fluoridation has greatly reduced the need for molar extractions. When treating Class II malocclusions that require the removal of teeth in the maxillary arch only, consideration should be given to the first molars as well as the more commonly extracted first premolars. This report describes such a case. The patient's active treatment ended in 1979, shortly before he left for college. After one retention visit, he was not seen again until he turned up in the early 1990s' literally on our doorstep, to practice general dentistry across the hall! PMID- 8712499 TI - The anterior alveolus: its importance in limiting orthodontic treatment and its influence on the occurrence of iatrogenic sequelae. AB - Delineating the limits of orthodontic treatment in nongrowing individuals is important when making treatment decisions, especially in borderline orthodontic surgical cases. The labial and lingual cortical plates at the level of the incisor apex may represent the anatomic limits of tooth movement. Cephalometric films of 107 adults were measured to determine the width of alveolar bone anterior and posterior to the incisor apex in each arch. Thin alveolar widths were found both labial and lingual to the mandibular incisors in groups of Class I, II, and III individuals with high SN-MP angle and in a group of Class III average SN-MP individuals. Thin alveolar widths were also found lingual to the maxillary incisors in a Class II high angle group. Clinical cases are presented showing that orthodontic tooth movement may be limited in patients with narrow alveolar bone widths and that these patients are likely to experience increased iatrogenic sequelae. PMID- 8712500 TI - Scintigraphic evaluation of 99mTc-methylene diphosphonate uptake in the navicular area of horses with lameness isolated to the foot by anesthesia of the palmar digital nerves. AB - OBJECTIVE: To evaluate distribution and intensity of 99mTc-methylene diphosphonate (99mTc-MDP) uptake in the navicular area in horses with forelimb lameness isolated to the palmar aspect of the foot. DESIGN: Prospective, case controlled study. ANIMALS: 7 horses with clinical signs of navicular syndrome and 7 control horses. PROCEDURE: Palmar view, soft tissue-phase scintigraphic images of the foot were obtained between 7 and 12 minutes after injection of 120 to 170 mCi of 99mTc-MDP. Lateral and palmar view, bone-phase images were obtained at 30 minutes and 1, 2, and 4 hours after injection. Palmar views were evaluated by determining the ratio of image density in the navicular area to mean image density in the distal phalangeal area. Palmar and lateral view, bone-phase images were also scored on the basis of navicular area intensity (intense = 3, moderate = 2, mild = 1, and no uptake = 0). Density ratios and mean scores were evaluated as a three-way ANOVA. RESULTS: Mean navicular-to-distal phalangeal density ratio for affected horses (1.77) was significantly (P = 0.003) greater than that for control horses (0.97). The mean subjective score for affected horses when evaluating palmar views only (1.85) and when evaluating palmar and lateral view pairs together (1.99) was significantly (P < 0.01) higher than scores for control horses (0.51, 0.62). Images obtained 1 hour after injection were as good at differentiating affected from control horses as images obtained between 2 to 4 hours after injection. CONCLUSION: A substantial number of horses with palmar foot pain have increased scintigraphic uptake within the navicular bone 1 to 4 hours after injection of 99mTc-MDP. Lateral view, bone-phase images are less sensitive than palmar view, bone-phase images in revealing navicular area uptake. CLINICAL RELEVANCE: A combination of lateral and palmar view scintigraphic images obtained between 1 and 4 hours after injection of 99mTc-MDP is a useful diagnostic aid in evaluating navicular bone involvement in horses with forelimb lameness isolated to the palmar aspect of the foot. PMID- 8712501 TI - Local distribution of mepivacaine after distal interphalangeal joint injection in horses. AB - OBJECTIVE: To evaluate the distribution of mepivacaine hydrochloride after distal interphalangeal (DIP) joint injection in horses. DESIGN: Prospective, uncontrolled study. ANIMALS: 10 adult horses. PROCEDURE: 30 minutes before euthanasia, 8 ml of 2% mepivacaine hydrochloride was injected into the dorsal pouch of a forelimb DIP joint. Synovial tissue from the DIP joint and podotrochlear (navicular) bursa and bone tissue from the medullary cavity of the distal sesamoid (navicular) bone were taken from both forelimbs immediately after death. All synovial and bone specimens were analyzed for tissue concentration of mepivacaine by high-performance liquid chromatography. Synovial tissue and bone specimen concentrations from the injected forelimb were compared with corresponding specimens from the noninjected forelimb. All synovial tissue and bone specimen concentrations were compared with an estimated effective tissue concentration of mepivacaine (0.3 microgram/mg) for local anesthesia. RESULTS: Specimen concentrations of mepivacaine from the injected forelimb were significantly greater (P < 0.05) than those in the corresponding tissues of the contralateral noninjected forelimb. All DIP joint and navicular bursa synovial tissue specimens from the injected forelimb had greater than the estimated effective tissue concentration of mepivacaine for local anesthesia. Of the 10 navicular bone specimens from the injected forelimb, 4 were higher and 2 were within 20% of the estimated effective tissue concentration of mepivacaine for local anesthesia. CONCLUSIONS: Mepivacaine hydrochloride deposited into the DIP joint should anesthetize pain arising from navicular bursa synovia and may decrease pain arising from the medullary cavity of the navicular bone. CLINICAL RELEVANCE: DIP joint injection of mepivacaine hydrochloride is not specific for DIP joint pain. PMID- 8712502 TI - Enzyme-linked immunosorbent assay for thrombin-antithrombin III complexes in horses. AB - OBJECTIVES: To adapt and characterize a human ELISA kit to quantify thrombin antithrombin III (TAT) complexes in horses, and to evaluate TAT as a marker for hypercoagulation in horses. ANIMALS: 29 clinically normal horses used as controls, and 4 ill horses used to evaluate assay for known causes of hypercoagulation. PROCEDURE: A commercially available human sandwich-type ELISA kit with 2 antibodies against human thrombin and antithrombin III that bind selectively to their corresponding TAT antigenic sites was used. Equine TAT standards were made from purified equine thrombin and antithrombin III. Proteins diluted in a phosphate-buffered saline solution containing 0.1% Tween and 1 U of heparin/ ml were used to establish standard curves. Reference intervals for TAT concentration in citrated equine plasma, and intra- and interassay coefficients of variation were determined. RESULTS: Mean +/- SD values were 3.95 +/- 1.93 micrograms/L, with median of 3.18 micrograms/L and range of 1.95 to 9.03 micrograms/ L. One horse with cecal perforation had TAT concentration of 174.30 micrograms/L, and a horse infused IV with endotoxin had TAT concentration of 62.98 micrograms/L 12 hours after infusion. CONCLUSIONS: The data suggest that human TAT ELISA kits can be used to measure TAT concentration in citrated equine plasma, and that TAT is a marker for hypercoagulation in horses. CLINICAL RELEVANCE: Assays for equine TAT many help to further characterize the hypercoagulable state in horses. PMID- 8712503 TI - Ultrasonography of the lungs, pleura, and mediastinum in healthy cows. AB - OBJECTIVE: To describe the ultrasonographic appearance of the lungs, pleura, and mediastinum of cattle. DESIGN: Transcutaneous ultrasonography of the lungs and pleura was performed on the right and left sides of the thorax. The mediastinum was examined transesophageally. ANIMALS: 50 healthy cows. PROCEDURE: The thoracic wall, pleura, and pulmonary surface were visualized on both sides of the thorax via transcutaneous ultrasonography. The cranial mediastinal lymph node, right azygos vein, aorta, and caudal vena cava were visualized via transesophageal ultrasonography. RESULTS: The layers of the thoracic wall appeared as narrow bends that varied in echogenicity. They were followed medially by an echogenic line that represented the costal and pulmonary pleurae. Reverberation artifacts were seen medial to the pleura. The pulmonary parenchyma could not be visualized ultrasonographically because of its air content. The cranial mediastinal lymph node appeared circular to oval. The right azygos vein was situated immediately adjacent to the cranial mediastinal lymph node. The aorta appeared circular to oval on cross-sectional view with an echogenic wall and an anechoic lumen. The caudal vena cava appeared as a tubular structure with an anechoic content and an echogenic wall. CONCLUSION: Ultrasonography of the lungs, pleura, and mediastinum in healthy cows provides information that can be used as a reference when examining cattle with suspected disease of the thorax. PMID- 8712504 TI - Immunohistolocalization of the carbonic anhydrase isoenzymes (CA-I, CA-II, and CA III) in the reproductive tract of male horses. AB - OBJECTIVE: To elucidate locations of cytosolic carbonic anhydrase isoenzyme (CA I, CA-II, and CA-III)-positive epithelial cells in equine male reproductive organs. DESIGN: Descriptive and immunohistochemical study. ANIMALS: 4 clinically normal male horses. PROCEDURE: The testis (seminiferous tubules, rete tubules), epididymis (initial, middle, and terminal segments), proximal and distal portions of the ductus deferens, ampulla ductus deferentis, seminal vesicle, prostate, and bulbourethral gland were excised from euthanatized horses after administration of an overdose of pentobarbital. The tissue specimens were quickly placed in fixative solution, dehydrated in ethanol, and embedded; then thin sections were cut. For immunohistochemical staining, antibodies against purified equine CA-I, CA-II, and CA-III were raised in rabbits. After examination of the specificity of each antiserum, the monospecific antisera against carbonic anhydrase isoenzymes were used to localize the isoenzymes. RESULTS: Specific staining for CA-III was found in the Sertoli and basal cells of the ductus deferens. Most of the testicular and epididymal tissue, as well as ductus deferens, were virtually negative for the enzymes when stained with the antibody to CA-I and CA-II. In the initial segment of the epididymis, a few principal cells had intense cytoplasmic staining with anti-CA-II. In the male accessory glands, CA-I, CA-II, and CA-III were detected in the epithelial cells of the seminal vesicle, prostate, and bulbourethral gland. CONCLUSIONS: In the equine male reproductive tract, the bicarbonate in semen originates mainly from accessory reproductive glands. All 3 isoenzymes may have central roles in the regulation of bicarbonate concentration in seminal plasm and, accordingly, regulate seminal plasma pH. Distribution of CA III in Sertoli and basal cells of the ductus deferens suggests other specialized physiologic roles. PMID- 8712505 TI - Rate of keratinization of the wall segment of the hoof and its relation to width and structure of the zona alba (white line) with respect to claw disease in cattle. AB - OBJECTIVES: To determine contribution of the wall segment of bovine cattle hoof to horn production, and relevance of structural differences of the wall segment and its horn production rate to claw disease. DESIGN: Epidermis and papillary body of the wall segment were examined by mesoscopy, light microscopy, and transmission and scanning electron microscopy. Morphometry of the entire length of the zona alba was examined, and the horn production rate of the wall segment was calculated. ANIMALS: Mixed-breed, dual purpose (beef and dairy) cattle of either sex, and young (20 months) Holstein-Friesian beef bulls. PROCEDURE: Blocks of a strip of the hoof from the coronary segment to the sole margin, including epidermis and dermis, were prepared for light and transmission electon microscopy. Prepared specimens of the wall-sole border were examined by scanning electron microscopy. Morphometry was performed on the outer, middle, and inner parts of the zona alba structures on unfixed horn specimens of beef bull claws. After removal of the zona alba specimens, the claw was removed and the proximodistal extent of the epidermal leaflets was measured and analyzed statistically. RESULTS: Horn production increased in the distal half of the wall segment, was greatest at the wall-sole border, and highest above the abaxial end of the zona alba. High horn production resulted in an incompletely keratinized, softer horn. CONCLUSIONS AND CLINICAL RELEVANCE: High horn production at the zona alba increases susceptibility to vascular disturbance. Claw dyskeratoses appear first in areas of high horn production, areas which are also subject to a greater frequency of claw lesions. PMID- 8712506 TI - Use of enzyme-linked immunosorbent assay to measure thrombin-antithrombin III complexes in horses with colic. AB - OBJECTIVES: To evaluate new ELISA for measurement of thrombin-antithrombin III (TAT) concentration, and to correlate the values to other tests of hemostasis in horses with colic. DESIGN: Plasma TAT concentration and 8 other hemostasis analytes were measured in horses with colic at hospital admission and during the next 4 days. Retrospectively, data were analyzed by outcome, broad-category diagnosis, and clinical management, and for correlation between TAT and other assays. ANIMALS: 100 horses with colic. PROCEDURE: Plasma samples were evaluated for TAT, fibrinogen, and fibrin degradation products concentrations; antithrombin III (ATIII), protein C, alpha 2-antiplasmin, and plasminogen activities; prothrombin time (PT); and activated partial thromboplastin time. RESULTS: Changes were indicative of a hypercoagulable state, most severe in nonsurviving horses, characterized by increased TAT concentration; decreased ATIII, protein C, and plasminogen activities; and increased PT. Nonsurvivors had significantly increased TAT concentration compared with that in survivors, without regard to sample collection time; however, compared over time, TAT was significantly increased only at admission. Highest TAT concentration was in nonsurvivors with inflammatory intestinal lesions. There was significant negative correlation between TAT and ATIII, protein C, alpha 2-antiplasmin, and plasminogen values, and significant positive correlation between TAT and PT, and fibrin degradation products values. CONCLUSIONS: Plasma TAT reflects the current state of coagulation system activation and is a good assay for early diagnosis of the hypercoagulable state in horses with the most severe forms of colic. CLINICAL RELEVANCE: Measurement of equine TAT provides further information to characterize the hypercoagulable state in horses to aid in case management. PMID- 8712507 TI - In vitro effect of ketones and hyperglycemia on feline hemoglobin oxidation and D and L-lactate production. AB - OBJECTIVES: To investigate in vitro effects of ketosis and hyperglycemia on feline erythrocyte Heinz body formation, reduced glutathione (GSH) concentration, and D- and L-lactate production, and to identify potential metabolic mechanisms of oxidative stress in diabetic cats. DESIGN: Washed feline erythrocytes suspended in buffers containing normal or high glucose concentration were incubated with various concentrations of ketone bodies and tested at defined time intervals for Heinz bodies, GSH concentration, and D- and L-lactate production. ANIMALS: Three healthy female domestic cats. PROCEDURE: Erythrocytes were washed, suspended in buffers containing 5 mM glucose (simulates euglycemia) or 25 mM glucose (simulates hyperglycemia), and incubated with acetone (5 and 10 mM, acetoacetate (5 and 10 mM), or beta-hydroxybutyrate (5 and 25 mM) for 24 hours at 37 C. Aliquots were stained with new methylene blue for Heinz bodies, and assayed spectrophotometrically for GSH and D- and L-lactate concentrations. Experiments were done in triplicate. Data were analyzed, using ANOVA with repeated measures. RESULTS: Neither high glucose concentration nor ketosis had direct effects on Heinz body formation or GSH values. Glutathione decreased to 89% of initial values over the 24-hour period in all samples. High glucose concentration also had no effect on erythrocyte D-lactate production; however, the rate of D-lactate production was slightly increased in samples containing 25 mM beta hydroxybutyrate. Linear L-lactate production confirmed metabolic viability of the erythrocyte suspensions. Samples in high glucose concentration produced L-lactate at a slightly higher (1.2x) rate than did samples in normal glucose concentration. CONCLUSIONS: High glucose concentration and ketosis do not account directly for oxidative damage and glyoxalase induction in feline erythrocytes in vitro, although high concentrations of beta-hydroxybutyrate may stimulate D lactate formation. PMID- 8712508 TI - Effects of vitamin E on mammary and blood leukocyte function, with emphasis on chemotaxis, in periparturient dairy cows. AB - OBJECTIVE: To determine the effect of vitamin E supplementation on the immune system of dairy cows. DESIGN: The following immune parameters were followed: production of chemotactic factors and superoxide by mammary macrophages and chemotactic responsiveness of blood neutrophils. ANIMALS: 16 healthy Holstein dairy cows. PROCEDURE: Dairy cows were assigned to 1 of 2 experimental groups: control (no vitamin E supplementation) and vitamin E supplemented. Supplementation of vitamin E started 4 weeks before and continued up to 8 weeks after parturition, and included oral supplementation of vitamin E at the rate of 3,000 IU/cow/d. In addition, the same group of cows received 1 injection of vitamin E (5,000 IU) 1 week prior to the expected date of parturition. Blood samples were collected weekly throughout the experimental period. RESULTS: Vitamin E supplementation enhanced by 30 to 83% (P < 0.05) chemotactic responsiveness of blood neutrophils beginning 2 weeks before to 4 weeks after parturition, compared with controls. There were no differences in production of superoxide or chemotactic factors by mammary macrophages between control and vitamin E-supplemented cows. CONCLUSIONS: Vitamin E supplementation prevents the periparturient inhibition of neutrophil chemotaxis. It is unlikely that vitamin E affects directly the function of mammary macrophages. PMID- 8712509 TI - Cytokine release by porcine livers perfused with lipopolysaccharide or live Salmonella choleraesuis. AB - OBJECTIVE: To develop a model to study the kinetics and relative amounts of cytokines produced by liver cells during enteric infection. DESIGN: Salmonella enteriditis lipopolysaccharide (LPS)- or live S choleraesuis-stimulated isolated livers from clinically normal pigs and pigs with active acute phase response. ANIMALS: 7- to 14-day-old salmonellosis-free pigs, 4 to 12/group. PROCEDURE: Livers were removed and perfused with oxygenated Krebs-Henseleit solution for 30 minutes and with S choleraesuis or LPS added for 7 minutes. Livers were then perfused with 500 ml of fresh solution in a closed loop procedure for 180 minutes. Perfusate samples were collected for tumor necrosis factor-alpha (TNF alpha) and interleukin 6 (IL-6) bioassays. RESULTS: Tumor necrosis factor-alpha values remained constant during perfusion of normal livers and increased in those exposed to LPS. Interleukin 6 values increased in perfusate from normal livers from 30 to 150 minutes, then decreased. In livers from pigs with an active acute phase response, TNF alpha values were reduced; IL-6 appeared by 2 minutes and decreased after 25 minutes. CONCLUSIONS: Isolated livers could be kept viable for 3 hours, and IL-6 and TNF alpha could be measured by the bioassays used. CLINICAL RELEVANCE: Model can be used for studying and modifying the response of liver cells to infective agents. PMID- 8712510 TI - Intramammary defense against infections induced by Escherichia coli in cows. AB - OBJECTIVE: To examine Escherichia coli lipopolysaccharide (LPS) effects on expression of CD14 and CD18 cell surface receptors and lectin/carbohydrate mediated nonopsonic phagocytosis of E coli. DESIGN: Cell isolation, monoclonal antibody, phagocytosis, and flow cytometric studies. ANIMALS: 4 clinically normal lactating Holstein cows for studies on CD14 and CD18, and 2 for phagocytosis studies. PROCEDURE: Binding of CD14 and CD18 monoclonal antibodies to blood and milk neutrophils and mononuclear leukocytes was studied by flow cytometry before and after intramammary injection of LPS, and nonopsonic phagocytosis of E coli by blood neutrophils was determined. Presence of intracellular CD14 was determined after in vitro incubation of neutrophils in skimmed milk and after fixation and permeabilization of freshly isolated neutrophils. RESULTS: Before LPS injection, percentages of blood neutrophils and large mononuclear (LMO) cells expressing CD14 averaged 3 and 63% and 68 and 35% for mammary neutrophils and LMO cells, respectively. After LPS injection, CD14 was only detected on blood and mammary LMO cells (61 and 25%); receptor expression increased by 1.8- and threefold, respectively. In vitro incubation of neutrophils in skimmed milk increased the percentage of neutrophils expressing CD14. The number of blood neutrophils staining positive for CD14 increased after permeabilization of the plasma membrane, which was blocked by unlabeled anti-CD14 monoclonal antibodies. Before LPS, percentages of blood neutrophils and LMO cells expressing CD18 averaged 93 and 95% and was 88 and 55% for mammary neutrophils and LMO cells, respectively. After LPS, percentages of mammary neutrophils and LMO cells expressing CD18 increased to 100 and 95%, respectively. Expression of CD18 was 2.6-fold higher for mammary neutrophils before injection of LPS, compared with blood neutrophils, either before or after LPS. In absence of opsonins, neutrophils with adherent and phagocytosed E coli averaged 83 and 14%. CONCLUSIONS: LPS modulated expression of CD14 and CD18 and lectin-carbohydrate interactions mediated nonopsonic phagocytosis of E coli. An intracellular pool of CD14 exists in bovine neutrophils and is capable of translocating to the cell surface. CLINICAL RELEVANCE: Development of methods to maximize expression of CD14 receptors on mammary neutrophils involved in production of tumor necrosis factor-alpha, and nonopsonic phagocytosis could result in reducing prevalence of mastitis in dairy cows. PMID- 8712511 TI - Serum neutralizing antibody response and protection against experimentally induced liver abscesses in steers vaccinated with Fusobacterium necrophorum. AB - OBJECTIVE: To determine the efficacy of leukotoxin-based Fusobacterium necrophorum vaccines and dietary tylosin in providing protection against experimentally induced hepatic abscesses in steers. DESIGN: 30 steers assigned randomly to 6 treatment groups of 5 steers each: 1, phosphate-buffered saline solution (PBSS; control); 2, PBSS control, fed tylosin (100 mg/steer) daily; 3, inactivated whole-cell culture with oil emulsion adjuvant; 4, culture supernatant (crude toxoid) with oil emulsion adjuvant; 5, semipurified leukotoxoid with oil emulsion adjuvant; and 6, semipurified leukotoxoid with saponin adjuvant. PROCEDURE: Steers were inoculated SC with emulsified antigen or PBSS on days 0 and 21. Blood samples were collected at weekly intervals to monitor serum antileukotoxin antibody titer. On day 42, all steers were challenge exposed intraportally with F necrophorum culture. Three weeks later (day 63), steers were euthanatized and necropsied to examine liver and assess protection. RESULTS: Antileukotoxin antibody titers of all vaccinated groups markedly increased from baseline values, and mean titers of vaccinated groups were higher than those of the control and tylosin-treated groups. Steers vaccinated with culture supernatant with oil emulsion adjuvant or semipurified leukotoxoid with saponin adjuvant had the highest mean antibody titers. All 5 steers in the control group developed liver abscesses. Tylosin feeding did not protect steers challenge exposed with F necrophorum intraportally. CONCLUSIONS: Culture supernatant was more protective than whole-cell culture or semipurified leukotoxin against experimentally induced hepatic abscesses. Partial purification of leukotoxin appeared to reduce its protective immunity. PMID- 8712512 TI - Salmonella enteritidis in eggs, cloacal swab specimens, and internal organs of experimentally infected White Leghorn chickens. AB - OBJECTIVE: To investigate prevalence of Salmonella enteritidis-positive eggs, excretion of the organism in fecal droppings, and infection of internal organs after oral inoculation of White Leghorn hens with S enteritidis phage type 8. ANIMALS: 30 White Leghorn laying hens. PROCEDURE: At 25 weeks of age, hens were each inoculated orally with 10(10) colony-forming units of S enteritidis, then were observed for 8 weeks. RESULTS: Salmonella enteritidis Y-BP2 did not cause any clinical signs of disease or decrease in egg production. However, at 1 week after inoculation, 63.9% of the eggs collected from inoculated hens were culture positive for S enteritidis. The organism was isolated from the shell washings, egg shells, yolk, and albumen. A total of 592 eggs from S enteritidis-inoculated hens were examined. Of these eggs, 157 (26.5%) were positive for S enteritidis on external shell washings alone, 17 (2.9%) were positive for S enteritidis internally, and 44 (7.4%) were positive for S enteritidis externally and internally. The percentage of culture-positive eggs gradually decreased between postinoculation weeks 2 and 5, then gradually increased to a high of 76% at week B. At 3, 7, and 10 days after S enteritidis inoculation, cloacal swab specimens from 3 hens were positive for S enteritidis. Salmonella enteritidis was recovered from ovary, oviduct, liver, and cecal junction from S enteritidis-inoculated hens. CONCLUSIONS: Our results indicated that birds infected with this isolate produced S enteritidis-positive eggs at high frequencies initially, that decreased over time. When S enteritidis antibody began to decrease, reaching geometric mean titer < or = 40, the frequency of S enteritidis-positive eggs increased. PMID- 8712513 TI - Hybridization of 2,659 Clostridium perfringens isolates with gene probes for seven toxins (alpha, beta, epsilon, iota, theta, mu, and enterotoxin) and for sialidase. AB - OBJECTIVE: To genetically characterize Clostridium perfringens isolates for association of pathologic type with various diseases. DESIGN: Prospective study. SAMPLE POPULATION: 2,659 C perfringens isolates from various nonhuman animals species, human beings, and foods. PROCEDURE: Colony hybridization with DNA probes for 7 toxin (alpha, beta, epsilon, iota (subunits a and b), theta, mu, and enterotoxin) genes and 1 sialidase gene were performed to group the isolates by pathologic type. RESULTS: Enterotoxin-negative type-A isolates were the most common (2,575/2,659), were isolated from all sources, and were separated into 5 pathologic types. In cattle and horses with enterotoxemia, essentially only these pathologic types were identified. The enterotoxin-negative isolates of types C or D each had a single pathologic type. Type-C isolates were isolated only from swine with necrotic enteritis and type-D isolates from small ruminants with enterotoxemia, except that 1 type-D isolate was also found from a healthy fish. Type-B or -E isolates were not found. Among the 47 enterotoxin-positive isolates, 5 isolates from sheep or deer were type D and the other 42 were type A. These 42 isolates were grouped into 3 pathologic types: 1 type was isolated from samples of almost all origins, but the other 2 types were found in only 5 fish, 4 human beings, and 1 dog. CONCLUSIONS AND CLINICAL RELEVANCE: Genetic characterization of these isolates allowed identification of 11 different pathologic types. This approach may be useful in molecular diagnosis and prophylaxis of clostridial disease. PMID- 8712514 TI - Impact of an orally administered insect growth regulator (lufenuron) on flea infestations of dogs in a controlled simulated home environment. AB - OBJECTIVE: To evaluate the ability of lufenuron to control cat flea (Ctenocephalides felis felis) populations on dogs under conditions simulating a naturally infested home environment. DESIGN: 2 treatment and 2 control groups of dogs. Treated dogs received lufenuron in tablet form monthly, and controls received excipient. Dogs had unrestricted access to indoor (carpeted) and outdoor (grassy) environments in which self-propagating flea populations had been established. ANIMALS: 17 adult female Beagles. PROCEDURE: Dogs were monitored for 77 days after initial infestation with fleas and 70 days after initial treatment. Efficacy of the drug was calculated on the basis of absolute reduction in flea counts and as a percentage of control. RESULTS: Lufenuron administration caused a statistically significant (P < 0.05) reduction in flea burdens in treated dogs, compared with controls. Initiation of treatment 7 days after infestation resulted in 75% control of F1-generation and 97% control of F2-generation fleas over a 70 day posttreatment period. CONCLUSIONS: Lufenuron was highly effective in reducing flea populations on dogs. The time required for control will vary with the duration (generation time) of the flea reproductive cycle and, hence, the geographic area in which the product will be used. The experimental results are most relevant to use of the product for control of an existing flea population in the Midwest. PMID- 8712515 TI - Myocarditis in mice and guinea pigs experimentally infected with a canine-origin Borrelia isolate from Florida. AB - OBJECTIVE: To characterize the pathogenic potential of a unique Borrelia isolate obtained from a dog from Florida (FCB isolate). DESIGN: Prospective experimental infection. ANIMALS: 32 preweanling Swiss Webster mice and 12 adult male Hartley guinea pigs were injected intraperitoneally with 10(5) spirochetes. PROCEDURE: Mice were used as controls and blood recipients, and at 3- to 4-day intervals, 1 control mouse and 2 infected mice were necropsied, tissues were cultured, and a recipient mouse was inoculated with blood. Guinea pigs were randomized to 4 groups and inoculated intradermally with 10(0), 10(2), 10(3), or 10(4) spirochetes. For 48 days, clinical, hematologic, serologic, and microbiologic tests were performed on them, after which they were necropsied. RESULTS: In mice, spirochetemia was detectable between postinoculation days (PID) 3 and 13, and seroreactivity to homologous antigen was detectable during PID 10 through 31. Compared with control mice, infected mouse spleens were 2 to 3 times larger. Histologic lesions included lymphoid hyperplasia, neutrophilic panniculitis, epicarditis, and myocarditis, with intralesional spirochetes detected from PID 3 through 6. During PID 10 through 31, nonsuppurative epicarditis developed. Signs of illness and hematologic abnormalities were not observed in guinea pigs, despite isolating spirochetes from blood during PID 7 to 27. When necropsied on PID 48, histologic lesions included lymphoid hyperplasia and lymphocytic plasmacytic epicarditis. CONCLUSIONS: The FCB isolate causes spirochetemia, lymphoid hyperplasia, dermatitis, and myocardial injury in Swiss Webster mice and can be transmitted by blood inoculation. In Hartley guinea pigs, the isolate causes spirochetemia, lymphoid hyperplasia, and epicarditis. Documentation of disease in mice, guinea pigs, and, presumably, dogs raises the level of concern that the FCB isolate might be pathogenic for man and other animal species. PMID- 8712516 TI - Evaluation of propofol for general anesthesia in premedicated horses. AB - OBJECTIVE: To evaluate selected hemodynamic, respiratory, and behavioral responses to propofol in horses premedicated with xylazine or detomidine. DESIGN: Xylazine (0.5 and 1.0 mg/kg of body weight) was administered IV on different days to each of 6 horses prior to IV administration of propofol (2 mg/kg). In a second group of 6 horses, detomidine (15 and 30 micrograms/kg) was similarly studied. ANIMALS: 2 groups of 6 mature healthy horses. PROCEDURE: Rectal temperature, heart and respiratory rates, arterial blood gas tensions, and direct arterial blood pressures were recorded before and at fixed intervals after drug administration. Induction and recovery events were quantitatively and qualitatively assessed. Cardiopulmonary and behavioral data to follow were statistically analyzed (P < or = 0.05). RESULTS: Heart rate decreased in dose dependent manner from a mean (+/- SD) of 39.5 +/- 5.1 beats/min after xylazine and detomidine. Second-degree atrioventricular dissociation was commonly seen at the higher drug doses. After propofol administration, heart rate either transiently increased or was less depressed early in recumbency, compared with predrug values. Direct arterial blood pressures varied inconsistently from predrug values. Mean arterial carbon dioxide tension tended to increase after drug administration (significance variable) from predrug values of 42 to 46 mm of Hg in both drug groups. After xylazine or detomidine administration, arterial oxygen tension decreased significantly from predrug values of 97 to 103 mm of Hg. The magnitude and duration of decrease was dose-dependent and greatest during recumbency. Behavioral responses to anesthetic induction were variable, but horses were uniformly calm and coordinated during recovery. Recumbency time increased in response to the higher dose of either premedicant drug. Mean (+/- SD) times to standing were 25.02 +/- 4.42 and 35.57 +/- 6.83 minutes for the low and high doses of xylazine, respectively and 41.04 +/- 11.21 and 52.64 +/- 14.67 minutes for the low and high doses of detomidine, respectively. CONCLUSION: Neither xylazine nor detomidine prevented excitation associated with propofol injection in horses. CLINICAL RELEVANCE: Xylazine or detomidine-propofol combinations likely will not replace common anesthetic induction techniques for horses. However, recovery characteristics associated with propofol encourage further study in horses. PMID- 8712517 TI - Disposition and excretion of 6-methoxy-2-naphthylacetic acid, the active metabolite of nabumetone in horses. AB - OBJECTIVE: To examine, in horses, the disposition and excretion of the active metabolite 6-methoxy-2-naphthylacetic acid (6MNA) of the nonsteroidal anti inflammatory prodrug nabumetone. DESIGN: Pharmacokinetic analysis of 6MNA after oral administration of nabumetone and IV administration of 6MNA. PROCEDURE: Using a crossover design, 5 horses were orally administered 3.7 mg of nabumetone/kg of body weight. After a 3-week washout period, 4 horses were administered 2.5 mg of 6MNA/kg, IV. RESULTS: Absorption of nabumetone from the gastrointestinal tract and its metabolism to 6MNA had a median appearance half-life of 0.88 hour. The elimination half-life was 11 hours. Area under the plasma concentration time curve for 6MNA after oral administration of nabumetone was 120.6 mg/h/L. A dose of 2.5 mg/kg of 6MNA administered IV resulted in plasma concentration nearly equivalent to that induced by the orally administered dose. Disposition of 6MNA was modeled as a one-compartment, first-order elimination. The area under the plasma concentration time curve for IV administration of 6MNA was 117.0 mg/h/L, and the specific volume of distribution was 0.247 L/kg. The distribution half life and the elimination half-life were 0.56 and 7.90 hours, respectively. Percentage of total dose recovered in urine for the 36-hour collection period after the oral and IV administrations was 7.4 and 5.3%, respectively. CONCLUSIONS: Metabolism of nabumetone to 6MNA, as reported in other species, also occurs in horses. There were a number of additional metabolites of nabumetone in urine that could not be fully identified and characterized. PMID- 8712518 TI - Evaluation of injections of collagenase and oxytetracycline via the umbilical artery as treatment for retained placenta in cattle. AB - OBJECTIVES: To test whether oxytetracycline inactivates collagenase when combined as a potential treatment for retained fetal membranes in cattle and to determine whether oxytetracycline passes to blood from fetal membranes after intraplacental injection. DESIGN: Prospective, controlled study. SAMPLE POPULATION: 288 placentomes from 12 cows in their third trimester of pregnancy and 4 cows at term pregnancy. PROCEDURE: 8 experimental groups were established: saline control, collagenase, collagenase plus oxytetracycline at 3 dosages, and oxytetracycline at 3 dosages. Placentomes were infused through an umbilical vessel with the test solutions and incubated at 39 C for 4 hours. Immediately after incubation, the force needed to detach cotyledons from caruncles was measured by a manometric technique. Cotyledon-caruncle interface fluids were analyzed for hydroxyproline (collagen breakdown) and total protein contents. A combination of collagenase and oxytetracycline was injected via umbilical arteries of cows undergoing cesarean section and in cows with retained fetal membranes after natural delivery. Antibiotic residue in blood was determined by the Bacillus stearothermophilus disk assay. RESULTS: There were no significant differences among collagenase and collagenase plus oxytetracycline groups in the amount of pressure needed to separate cotyledon from caruncle, amount of hydroxyproline released, and amount of total protein broken down. The 4 cows tested negative for oxytetracycline in the blood. CLINICAL RELEVANCE: Oxytetracycline and collagenase may be a potential combination treatment for retained fetal membranes in cattle. In addition, the lack of antibiotic residue detection in blood may be of regulatory relevance. PMID- 8712519 TI - Efficacy of florfenicol for treatment of clinical and subclinical bovine mastitis. AB - OBJECTIVE: To evaluate efficacy of florfenicol treatment for bovine mastitis caused by Streptococcus agalactiae, Staphylococcus aureus nonagalactiae streptococci, coagulase-negative staphylococci, Escherichia coli, Klebsiella sp, and others. DESIGN: Double blind study with cases randomly assigned to 1 of 2 treatment groups. SAMPLE POPULATION: 861 cows/10 commercial dairy farms. PROCEDURE: Experimental (750 mg of florfenicol) or control (200 mg of cloxacillin) treatment was administered by intramammary infusion every 12 hours for 3 treatment to all cases. Treatments were randomly assigned identified only by numerical labels. To retain blinding, the longer withdrawal time was adhered to for all cases. Cases remained in the study only if there was no other treatment. Quarter samples were recultured 14, 21, and 28 days later. If all samples after day 1 were culture negative, the case was defined as cured. If only 1 of the follow-up results was positive, the case was considered cured if the day 28 somatic cell count was < 300,000/ ml. Failure of treatment was defined as 2 or more culture positive follow-up samples. RESULTS: Florfenicol and cloxacillin did not differ significantly in efficacy versus clinical (n = 85) or subclinical (n = 71) bovine mastitis, or for any etiologic agent (X2). Overall cure rates for mastitis were: Str agalactiae, 5 of 8 (63%); Sta aureus, 5 of 54 (9%); Streptococcus sp, 16 of 35 (46%); Staphylococcus sp, 7 of 33 (21%); E coli, 5 of 11 (46%); Klebsiella sp, 3 of 6 (50%); others, 1 of 9 (11%); and all cases, 42 of 156 (27%). CONCLUSIONS: Florfenicol did not offer any advantage over cloxacillin in efficacy against bovine mastitis. Overall cure rates were low. As with most mastitis treatment regimens poor efficacy may be partly attributable to the short duration of treatment. PMID- 8712520 TI - Anesthesia induced in pigs by use of a combination of medetomidine, butorphanol, and ketamine and its reversal by administration of atipamezole. AB - OBJECTIVE: To develop an IM administrable anesthetic combination for pigs. DESIGN: Use of a combination of atropine, medetomidine, butorphanol, and ketamine (MB-K) was evaluated as an anesthetic regimen and compared with that of a combination of atropine, xylazine, butorphanol, and ketamine (XB-K). Cardiorespiratory effects of MB-K combination and use of atipamezole as a means of reversing anesthesia induced by MB-K were examined. ANIMALS: 18 castrated, mixed-breed, specific-pathogenfree pigs, aged 8 to 15 (mean, 12.1) weeks and weighing 14.5 to 26.0 (mean, 19.6) kg. were studied. PROCEDURE: Dosages of drugs used in this study were atropine, 25 micrograms/kg of body weight; medetomidine, 80 micrograms/kg; xylazine, 2 mg/kg; butorphanol, 200 micrograms/kg; ketamine, 10 mg/kg; and atipamezole, 240 micrograms/kg. RESULTS: MB-K combination proved to be more effective than XB-K combination as an anesthetic combination. After quick and smooth induction by IM administration, MB-K-induced anesthesia was sustained for 98.8 +/- 22.5 minutes (mean +/- SD, 47.4 +/- 16.5 minutes by XB-K) with accompanying muscular relaxation (91 +/- 18 minutes) and loss of pedal (82 +/- 24 minutes) and laryngeal (75 +/- 19 minutes) reflexes. Loss of these reflexes was of significantly longer duration than the loss induced by XB-K, enabled tracheal intubation, and, thus, supported major surgery for at least 30 minutes after induction. Recovery from MB-K-induced anesthesia was smooth. MB-K combination had a slight stimulative effect on cardiovascular status, and a significant depressant effect on blood gas and acid-base status, but these effects were within biologically acceptable limits. Oxygen consumption of pigs under MB-K induced anesthesia decreased significantly. MB-K-induced anesthesia could be effectively and quickly reversed by IM or IV administration of atipamezole. CONCLUSIONS: The combination of medetomidine, butorphanol, and ketamine induces excellent surgical anesthesia in pigs, and results in moderate cardiorespiratory effects. A great advantage of the anesthetic regimen is that it can be effectively and quickly reversed by atipamezole. CLINICAL RELEVANCE: Medetomidine, butorphanol, and ketamine-induced anesthesia is available for short term major surgery in pigs. PMID- 8712521 TI - Comparison of sedative and cardiorespiratory effects of medetomidine and medetomidine-butorphanol combination in dogs. AB - OBJECTIVE: Sedative, cardiorespiratory, and analgesic effects of intramuscular administration of medetomidine (40 micrograms/kg of body weight)-glycopyrrolate (0.01 mg/kg) and medetomidine (10 micrograms/kg)-butorphanol (0.2 mg/kg) glycopyrrolate (0.01 mg/kg) combinations were compared. Additional evaluations were done on reversal of medetomidine, using atipamezole (200 micrograms/kg. IV), after 90 minutes of medetomidine-induced sedation. DESIGN: Crossover study, with each dog receiving each drug combination at 1-week intervals. ANIMALS: Six 2-year old English hound-type dogs. PROCEDURE: Arterial blood pressure, ECG, respiratory rate, tidal volume, minute volume, arterial blood gas tensions, and serum biochemical variables were measured before, during, and after sedation. Analgesia was evaluated by needle prick on the skin and tail clamp. RESULTS: Heart rate decreased significantly from 100 beats/min to < 40 beats/min within 3 minutes of injection of medetomidine and medetomidine and butorphanol (MB). Mean arterial blood pressure in both groups were maintained above 100 mm of Hg throughout the recording period. There was no significant difference between medetomidine and MB in respiratory rate, tidal volume, and minute ventilation. Hypoxemia (PaO2 < 60 mm of Hg) was observed at 10 and 20 minutes in 2 dogs given MB. Atipamezole administration in the dogs given medetomidine significantly increased PaO2, and returned the values to baseline. Needle prick analgesia duration was longer in the medetomidine (80 +/- 7.7 minutes) than MB (56.0 +/- 19.2 minutes) group. Tail pinch analgesia was variable in both groups. Duration of lateral recumbency was longer after medetomidine (90 +/- 0 minutes) than MB (73.5 +/- 19.0 minutes). CONCLUSION: Medetomidine and MB were effective combination for mildly invasive procedures. CLINICAL RELEVANCE: MB induced a shorter period of analgesia and recumbency than did medetomidine. PMID- 8712522 TI - Effects of cisapride on feline colonic smooth muscle function. AB - OBJECTIVE: To determine the effects of cisapride on feline colonic smooth muscle function. DESIGN: In vitro smooth muscle mechanical measurements. ANIMALS: Intact colon was obtained from healthy 2- or 3-year-old cats. PROCEDURE: Longitudinal smooth muscle strips from proximal and distal portions of feline colon were suspended in physiologic buffer solution (37 C. 100% O2, pH 7.4), attached to isometric force transducers, and stretched to optimal muscle length. Control responses were obtained at each muscle site with acetylcholine (10(-8) to 10(-4) M), cholecystokinin (10(-11) to 10(-7) M), substance P (10(-21) to 10(-7) M), or neurotensin (10(-11) to 10(-7) M). Muscles were then stimulated with cumulative (10(-9) to 10(-6) M) or bolus (10(-6) M) doses of cisapride in the absence or presence of tetrodotoxin (10(-5) M) and atropine (10(-6) M), nifedipine (10(-6) M), or calcium-free buffer solution. RESULTS: Cisapride stimulated contractions of longitudinal smooth muscle from proximal and distal portions of feline colon that were similar in magnitude to contractions induced by substance P and neurotensin. Cisapride contractions were only partially inhibited by tetrodotoxin (10(-6) M) and atropine (10(-6) M), suggesting that cisapride responses are only partially dependent on enteric cholinergic nerves. Nifedipine (10(-6)M) inhibited the maximal contraction to cisapride (10(-6) M) by approximately 80%. Removal of extracellular calcium did not inhibit cisapride contractions to a greater extent than did inhibition by nifedipine, indicating that calcium influx through voltage dependent calcium channels was predominantly responsible for the dependence of the cisapride contraction on extracellular calcium. CONCLUSIONS: Cisapride induced contractions of feline colonic smooth muscle are largely smooth muscle mediated and dependent on calcium influx, and are only partially dependent on enteric cholinergic nerves. CLINICAL RELEVANCE: Cisapride may be useful in the treatment of feline colonic motility disorders. PMID- 8712523 TI - Penetration of enrofloxacin and ciprofloxacin into breast milk, and pharmacokinetics of the drugs in lactating rabbits and neonatal offspring. AB - OBJECTIVE: To determine the pharmacokinetics and milk penetration of enrofloxacin (ENR) and ciprofloxacin (CIP) in lactating rabbits and their disposition in suckling rabbits. DESIGN: Prospective cross-over study. ANIMALS: 6 lactating New Zeland White rabbits and their offspring (16 days after parturition). PROCEDURE: Serial plasma and milk samples were assayed by use of a high-performance liquid chromatography technique. In vitro protein binding in plasma and skim milk was measured by ultrafiltration. Skim-to-whole milk ratio also was determined. The time course of ENR and CIP was fitted by nonlinear least squares regression analysis, and the pharmacokinetic variables were compared. RESULTS: The time courses of ENR and CIP in plasma were similar in lactating adult rabbits (mean body clearances, 23.9 and 27.2 ml/min/kg of body weight, for ENR and CIP, respectively). Observed milk-to-plasma ratios (M/P) were determined, using the area under the milk and plasma concentration versus time profiles (ENR, 2.59; CIP, 3.61). Predicted M/P (ENR, 6.35; CIP, 3.04) were calculated from in vitro measurements. Body clearance calculated for ENR and CIP in suckling rabbit pups involved a decrease of 80 and 74%, respectively, over that found in lactating animals. CONCLUSIONS: Observed CIP M/P were correlated to predicted values, which strengthens the argument that CIP passes into the milk by nonionic diffusion. The lack of correlation between observed and predicted ENR M/P pointed out that ENR undergoes faster elimination from milk than that predicted by the diffusional model. Diminished elimination capacity observed in suckling rabbits would result in greater exposure than that predicted from concentrations alone. PMID- 8712524 TI - Regulation of glycosaminoglycan metabolism by bone morphogenetic protein-2 in equine cartilage explant cultures. AB - OBJECTIVE: To investigate whether recombinant human bone morphogenetic protein-2 (rhBMP-2) regulates glycosaminoglycan (GAG) synthesis and release from equine articular cartilage explant cultures. DESIGN: Equine articular cartilage explants were maintained in vitro for 7 days in the presence of 0 (control), 1, 10, or 100 ng of rhBMP-2/ml. Synthesis and release of GAG were assessed as measures of production and degradation of the extracellular matrix, respectively. ANIMALS: 6 horses (age range, 2 to 25 years old) without clinically detectable musculoskeletal abnormalities. PROCEDURE: Rate of synthesis of GAG was assessed by incorporation of [36S]sulfate during the final 24 hours of the 7-day incubation period. Release of GAG was assessed on days 3, 6, and 7, using 1,9 dimethylmethylene blue. RESULTS: Explants from all 6 horses had a significant (P = 0.05) increase in release of GAG in response to incubation with 100 ng of rhBMP 2/ml. There was a significant (P = 0.05) decrease in GAG synthesis in explants from only 2 of the 6 horses at the same concentration of rhBMP-2. There was no significant age correlation between responsive and nonresponsive horses. CONCLUSIONS: A concentration of 100 ng of rhBMP-2/ml stimulates GAG release from explant cultures of equine articular cartilage. The data suggest that bone morphogenetic proteins may be potential regulators of equine cartilage degradation and repair. CLINICAL RELEVANCE: Surgical procedures that damage subchondral bone may stimulate generation of improved cartilage-like tissue. It is, therefore, crucial to understand how bone-derived factors may influence cartilage metabolism in horses. PMID- 8712525 TI - Effects of oral lactose and xylose loads on blood glucose, galactose, xylose, and insulin values in healthy calves and calves with diarrhea. AB - OBJECTIVE: 2 hypotheses were tested: calves with acute, mild diarrhea digest lactose less efficiently than healthy calves, and they are in a catabolic state, which influences plasma glucose concentration after glucose absorption. DESIGN: Clinical study; 2 treatments with 10 repetitions/treatment. ANIMALS: 20 preruminant Brown Swiss and Simmental Red Holstein calves; 10 calves with mild diarrhea, and 10 age-matched healthy calves. PROCEDURE: Blood metabolite and hormone concentrations were determined before and after an oral lactose load. Plasma xylose concentration was determined after an oral xylose load. III calves were tested 1 day after the onset of diarrhea. RESULTS: Calves with diarrhea (cryptosporidia, coronavirus) had lower preprandial concentrations of plasma glucose, insulin-like growth factor 1, and 3,5,3-triiodothyronine (P < 0.01) and a higher concentration of free fatty acids (P < 0.03) than did healthy calves. After the oral lactose and xylose loads, blood galactose and plasma xylose concentrations were lower in ill calves (P = 0.10 and P = 0.07, respectively). In calves with diarrhea, there was a larger increase of plasma glucose concentration (P = 0.12) and a smaller increase of plasma insulin concentration (P = 0.04) above baseline values after lactose ingestion. CONCLUSIONS: Lactose digestion is slightly impaired in calves with mild diarrhea. Calves with acute diarrhea are in a catabolic state and, therefore, respond with a larger increase of plasma glucose concentration to a given amount of absorbed glucose than do healthy calves. CLINICAL-RELEVANCE: Plasma glucose concentration is not a reliable measure for glucose absorption in animals that are in a catabolic state. PMID- 8712526 TI - Mechanisms of acid injury in porcine gastroesophageal mucosa. AB - OBJECTIVE: To identify the cause and mechanisms of injury in gastroesophageal ulcer disease in market weight swine. DESIGN: Comparison of mechanisms of injury caused by HCI with those caused by short chain fatty acids (SCFA) in gastric mucosa. ANIMALS: Pigs weighing 30 to 40 kg. PROCEDURE: Gastric tissues were studied in Ussing chambers; short-circuit current (lsc) and electrical resistance (R) were recorded in response to treatment, and tissues were examined histologically. RESULTS: 60 mM mucosal acetate abruptly ( < or = 75 minutes) and irreversibly abolished lsc at pH < or = 4.5, whereas R decreased more slowly. These data were associated with cell swelling and vesicle formation in mid-zonal layers, followed by sloughing of the outer barrier, erosion into deeper zones, and finally, ulceration. Mucosal HCl at pH > 1.5 was ineffective; however, at pH 1.5, HCl induced an abrupt decrease in R, followed by a slow decrease in lsc, an effect opposite to that caused by SCFA. Serosal addition of HCl rapidly abolished lsc suggesting a barrier to free H+ diffusion from the mucosal solution. CONCLUSIONS: Undissociated SCFA rapidly penetrate the outer barrier and acidify underlying viable tissue. Cellular acidification inhibits Na pumping and osmoregulation, resulting in cell swelling and necrosis. In contrast, HCl induces and increase in outer barrier permeability before accessing the transporting cells, a much longer process ( > or = 5 hours) requiring a lower pH. These studies suggest that microbial production of SCFA may be important in the pathogenesis of porcine gastric ulcers. PMID- 8712528 TI - Alterations in colonic smooth muscle function in cats with idiopathic megacolon. AB - OBJECTIVE: To determine whether colonic smooth muscle dysfunction is involved in the pathogenesis of idiopathic megacolon in cats. DESIGN: In vitro smooth muscle mechanical measurements. ANIMALS: Colon from healthy cats and cats with idiopathic megacolon. PROCEDURE: Colonic smooth muscle strips were suspended in physiologic buffer solution, attached to isometric force transducers, and contracted with acetylcholine (ACh; 10(-9) to 10(-4)M), substance P (SP; 10(-10) to 10(-6)M), cholecystokinin (CCK; 10(-11) to 10(-8)M), potassium chloride (KCl; 10 to 80 mM), or electrical field stimulation (EFS; 25 V, 1 to 30 Hz, 0.5 millisecond duration). Isometric stress responses were compared with those obtained from healthy controls. Colonic smooth muscle strips were also evaluated histologically for neuronal and smooth muscle cell morphology. RESULTS: Passive isometric stress was not altered, but the active isometric stress responses of megacolon smooth muscle to ACh, SP, CCK, KCl, and EFS were significantly (P < 0.05) diminished, compared with healthy controls. Differences were observed in longitudinal and circular smooth muscle from proximal and distal portions of the colon. Histologic evaluation revealed few abnormalities of smooth muscle cells or of myenteric or submucosal plexus neurons. The contractile response of megacolon smooth muscle to EFS, and the inhibition of this response by tetrodotoxin, suggest that myenteric and submucosal plexus neurons in megacolon smooth muscle are functional. CONCLUSIONS: Idiopathic megacolon is a generalized dysfunction of colonic smooth muscle in cats. The diminished isometric stress responses to receptor occupancy (ACh, SP, and CCK) and membrane depolarization (KCl) further suggest that the disorder involves disturbance in the activation of smooth muscle myofilaments. PMID- 8712527 TI - Effect of pertussis toxin and B-oligomer on platelet-activating factor-induced generation of inositol phosphates in porcine alveolar macrophages. AB - OBJECTIVE: To test the hypothesis that platelet-activating factor (PAF) induces inositol phosphate turnover through a receptor-linked, pertussis toxin-sensitive guanine nucleotide-binding (G) protein-dependent pathway in porcine alveolar macrophages. DESIGN: Randomized complete block design was used with 2 or 3 replicates/block. ANIMALS: Porcine alveolar macrophages were obtained by lavage of excised lungs from Yorkshire-type pigs (mean +/- SEM, 21 +/- 2 kg). PROCEDURE: Phospholipase C activation was assessed, using anion exchange chromatography to measure accumulation of inositol phosphates in [3H]myo-inositol-labeled alveolar macrophages. Macrophages were incubated with saline solution, pertussis toxin (4.75 nM), or B-oligomer (4.75 nM) for 2 hours. Cells then were washed and incubated for 5 minutes with PAF (0, 0.1, 1.0, or 10 microM; n = 15). Results were expressed as total inositol phosphates (inositol monophosphate, bisphosphate, trisphosphate, and tetrakisphosphate). RESULTS: Concentrations of total inositol phosphates were significantly (P < 0.05) increased to 162 +/- 7, 172 +/- 4, and 194 +/- 9% of control in response to 0.1, 1.0, and 10 microM PAF, respectively. Pertussis toxin attenuated the PAF-induced increase in total inositol phosphates by approximately 50% (P < 0.05). The B-oligomer of pertussis toxin failed to modify PAF-induced increases in total inositol phosphates. The specific PAF receptor antagonist WEB 2086 markedly attenuated PAF-induced. (10 microM) increase in inositol phosphates. CONCLUSIONS: We conclude that PAF stimulates accumulation of inositol phosphates through a specific receptor and that a pertussis toxin-sensitive G protein is involved in the signal transduction process leading to activation of phospholipase C in porcine alveolar macrophages. PMID- 8712529 TI - Surgical creation of portacaval shunts during temporary intestinal arterial and venous occlusion in dogs. AB - OBJECTIVE: To surgically create complete portacaval shunts in dogs during temporary arrest of intestinal arterial and portal venous blood flow. DESIGN: Complete portacaval anastomoses were surgically created, and liver function was evaluated for 14 to 18 weeks after surgery. ANIMALS: 32 adult mixed-breed dogs of either sex. PROCEDURE: Administration of deferoxamine and temporary intestinal arterial occlusion were used to minimize the intestinal cellular damage resulting from the complete, temporary arrest of portal venous blood flow during creation of the portacaval anastomosis. Side-to-side, appositional anastomoses ( > 2 cm diameter) were formed between the portal vein and caudal vena cava. Dogs were observed daily for signs of hepatic encephalopathy, and food intake was recorded. Body weight was recorded weekly. Preprandial plasma ammonia, serum urea nitrogen, and glucose concentrations and sulfobromophthalein retention were measured monthly. The dogs were euthanatized, and necropsy was performed 14 to 18 weeks after surgery. RESULTS: 30 of 32 dogs recovered without complications. Complete portosystemic shunting was documented by increased plasma ammonia concentration, decreased serum urea nitrogen and glucose concentrations, prolonged sulfobromophthalein retention (P < 0.01), and inspection at necropsy. CONCLUSION: This method of providing temporary, complete arrest of portal venous blood flow was helpful in allowing accurate, appositional portacaval anastomoses to be created that remained patent for 14 to 18 weeks. CLINICAL RELEVANCE: This method of providing temporary, complete arrest of portal venous blood flow may prove useful in clinical surgery when temporary arrest of portal blood flow is desired. PMID- 8712530 TI - Nurses score victory in NLRB decision: ANA secures protections for RNs. PMID- 8712531 TI - ANA asserts attacks on practice threaten patient safety: nursing's response seeks to protect patients, preserve practice. PMID- 8712532 TI - Study links rotating shift work and nurses' risk of coronary heart disease. PMID- 8712533 TI - Implications of the past year on the upcoming election. PMID- 8712534 TI - Obstetric nursing. Field offers new opportunities. PMID- 8712535 TI - Articulation: Pennsylvania's solution to educational mobility in nursing. PMID- 8712536 TI - Blizzard '96: nurses undaunted by record snowfall. PMID- 8712537 TI - Oncology nursing. Practice blooms to meet growing need. PMID- 8712538 TI - Nursing must be strengthened by its diversity. PMID- 8712539 TI - Nurse staffing and the problem of lack of data. PMID- 8712540 TI - Nursing unity needed to preserve practice. As I see it. PMID- 8712542 TI - IOM issues nursing staffing report: some positive recommendations yet report fails to address immediacy of hospital staffing problems. PMID- 8712541 TI - Incentive programs for nurses surfacing: does practice undermine nurse-patient bond? PMID- 8712543 TI - Nurses become political "tour de force". PMID- 8712544 TI - ANA posts big wins in contentious [challenging 104th Congress]. PMID- 8712545 TI - Nurse ranks grow in legislative bodies nationwide. PMID- 8712546 TI - ANA talks to the presidential candidates. Interview by Chris deVries. PMID- 8712548 TI - Labor groups strive to protect workers in growing managed care era. PMID- 8712547 TI - SNAs face threats to practice, push quality initiatives. PMID- 8712549 TI - Critical care nursing moves into home, community. PMID- 8712550 TI - ANA calls workplace guidelines an "excellent step". PMID- 8712551 TI - Steps taken to protect health care workers from violence. PMID- 8712552 TI - ANA volunteers: say thanks and join their ranks! PMID- 8712553 TI - Rx for change: grass-roots involvement in the political process as needed. As I see it. PMID- 8712555 TI - Basic behavioral science research for mental health. Social influence and social cognition. Basic Behavioral Science Task Force of the National Advisory Mental Health Council. PMID- 8712554 TI - Staff nurse involvement in SNAs crucial to addressing profession's concerns: winter summit taps RNs' leadership abilities. PMID- 8712556 TI - The shifting gender composition of psychology. Trends and implications for the discipline. AB - Psychology, along with the majority of professions and scientific disciplines, has undergone dramatic shifts in gender composition over the past two decades. These changes have prompted concern that this increased participation by women may lead to erosion in the status of these occupations. This article describes the results of a case study of psychology conducted by a subcommittee of the American Psychological Association's (APA's) Task Force on the Changing Gender Composition of Psychology to examine the discipline's changing gender composition and the factors related to these shifts. Societal and disciplinary trends are examined, along with data on the patterns of men's and women's involvement in the educational pipeline and workplace. The results provide little support for the concern over the increasing representation of women and its impact on the prestige of the discipline. Rather, they suggest that changes in the nature and status of psychology per se may be at least partly responsible for the changes in male and female participation and that the nature, magnitude, and causes of these disciplinary changes require further examination. Specific recommendations for the APA prepared by another subcommittee of the Task Force are also presented in the Appendix. PMID- 8712558 TI - Open versus laparoscopic surgery: a comparison of natural antitumoral cellular immunity in a small animal model. AB - The scope of laparoscopic surgery has extended to the treatment of cancer. The immunological impact of laparoscopic surgery as compared with open surgery has not been well characterized. A paucity of information is available about differences or similarities of these two methods regarding natural antitumoral cellular immunity, namely, natural killer cell cytotoxicity. This study compared the activity of natural killer cells in rats subjected to pneumoperitoneum, open dissection of the retroperitoneum, and laparoscopic dissection of the retroperitoneum. When compared to control animals, rats subjected to pneumoperitoneum did not show any change in natural killer cell activity. Conversely, the groups of open surgery and laparoscopic surgery revealed significantly decreased natural killer cell cytotoxicity compared with controls (P < 0.0167). When the laparoscopic and the open surgical groups were compared to each other, no difference was found. In this study, both open and laparoscopic surgery had a suppressive effect upon the natural antitumoral cellular immunity. Pneumoperitoneum did not have an immune suppressive effect on natural killer cell activity. In this model, the advantages of laparoscopic surgery do not apply to natural antitumoral cellular immunity. PMID- 8712557 TI - Urokinase expression in transduced endothelial cells. AB - Increased thromboresistance through the release of lytic agents by endothelial cells may improve the patency of endothelial lined prosthetic grafts. We have evaluated the expression of urokinase from cells transduced with a retrovirus containing the gene for a human preprourokinase. Endothelial cells were enzymatically harvested from canine external jugular vein in nine animals and grown to confluence in culture. One-third of these cells served as controls, and the remaining two-thirds were transduced via incubation with an LXSN-type retroviral vector carrying the urokinase gene and a neomycin resistance gene. Successfully transduced cells were selected by incubation with 400 micrograms/mL G418 and pure cultures grown to confluence. Supernatants from confluent control and experimental cell cultures after 48 hours in defined, serum-free medium were assayed for human urokinase concentration and overall enzyme activity. ELISA quantitation of concentration using mouse antihuman urokinase antibody showed 0.15 +/- 0.11 ng/mL/hr/10(6) cells in the transduced cell supernatant; no measurable concentration was found in the control cells. (P < 0.01) Overall (human plus canine) enzyme activity of urokinase was determined using an indirect spectrophotometric assay based on plasminogen activation (ploug U/mL). Transduced cells showed activities of 0.12 at 10 days and 0.45 at confluence; control cell activity was 0.0 and 0.15, respectively. (P < 0.05) These data show that endothelial cells can be transduced with a urokinase expressing gene that increases the release of this thrombolytic agent. Lining small diameter prosthetic grafts with these cells may improve their thromboresistance and long term patency. PMID- 8712559 TI - Laparoscopic fundoplication: a three-year review. AB - Laparoscopic treatment of gastroesophageal reflux disease provides decreased postoperative pain and shortens hospitalization. Experience with this procedure continues to grow. The charts of patients who had laparoscopic fundoplication for the treatment of symptomatic refractory gastroesophageal reflux over a three year period were reviewed. Age, sex, preoperative evaluation, surgical technique, length of procedure, length of hospitalization, time to regular diet, conversion to laparotomy, complications, and long-term outcome were analyzed. Eighty-eight patients, 39 females and 49 males, underwent laparoscopic gastric fundoplication. All patients has symptomatic gastroesophageal reflux. Preoperative evaluation including upper endoscopy demonstrated esophagitis in 71 patients (81%) with changes of Barrett's esophagitis in 12 patients (14%). Decreased lower esophageal sphincter pressures were found in 33 of 73 patients (45%) and none had abnormal motility identified. A 2-3 cm fundoplication with nonabsorbable suture was performed over a large dilator. Mean operative time was 109 minutes. The mean length of hospitalization was 1.9 days in patients completed laparoscopically and six days in the 3 patients that required conversion to celiotomy for completion of their fundoplication. A clear liquid diet was offered the evening of surgery and advanced to regular as tolerated. Mild dysphagia was found in most patients for the first week and full liquid or soft diet was prescribed. All but seven patients advanced to regular diet by two weeks. Operative complications occurred in four patients and included two pneumothoraces and two esophageal perforations. Follow-up to 40 months has identified one partial wrap disruption and one crural breakdown both repaired laparoscopically. Seventy-seven patients (88%) have remained asymptomatic and off all medication. Laparoscopic fundoplication is safe and effective, allowing the benefits of minimal access surgery, including decreased postoperative pain and disability. PMID- 8712560 TI - Will stereotactic breast biopsy achieve results as good as current techniques? AB - Stereotactic procedures recently have been advocated to replace most needle localization and open biopsy procedures. In order to provide a baseline for comparison at our institution, a retrospective review of our results over the last 3 years was performed. During this time period, 496 biopsies were performed in 480 patients. Needle localization was done in 311 cases, whereas the remaining 185 biopsies were done for palpable masses. There were no significant differences in either the positive rate (19.0% vs 13.5%) or the infection rate (2.6% vs 1.6%) in the two groups. Follow-up of all patients has revealed no missed carcinomas and no referrals to a plastic surgeon for a poor cosmetic result. Current breast biopsy techniques yield good results, with acceptably low morbidity rates. Given that approximately one in five needle localization biopsies detects a malignancy, a negative result following a stereotactic biopsy may not preclude a needle localization procedure. It is therefore unlikely that stereotactic procedures will lead to an overall decrease in health care costs. Surgeon involvement will be crucial to assure best and most cost-effective results. PMID- 8712561 TI - Failure of antiseptic bonding to prevent central venous catheter-related infection and sepsis. AB - Infection associated with the use of triple lumen catheters in hospitals is a frequent and serious complication. The prevailing hypothesis for the origin of catheter-related infection (CRI) is bacterial colonization and subsequent infection of the skin insertion site and catheter interface. The recently released ARROWgard catheter contains a bonded synergistic combination of silver sulfadiazine and chlorhexidine, which is thought to render the catheter resistant to bacterial colonization and subsequent sepsis. The purpose of this study is to compare the incidence of CRI and catheter-related sepsis (CRS) between a standard triple lumen catheter and ARROWgard antiseptic coated catheter in patients receiving total parenteral nutrition (TPN). A randomized, prospective clinical trial was conducted at a community referral center from January 1993 through April 1994. One-hundred-ninety-one patients with need for TPN were randomized to receive either the ARROWgard or a standard triple lumen catheter placed under a strict sterile protocol. CRI was defined as >/= 15 colony forming units by semiquantitative culture technique of the catheter tip or intracutaneous segment. CRS was defined as growth of the same organism on the catheter and at least one peripheral blood culture. All catheters were cultured. Ninety-two patients received the ARROWgard catheter, and 99 patients received the standard catheter. There were no differences between the average age, sex, length of hospital stay, days on TPN, number of catheters/patient, indications for TPN, primary diagnoses, or duration of the central line between the two groups. The overall rate of CRI was 11.5 per cent, and CRS was 8.4 per cent in this study. The rate of CRI for the ARROWgard was 10.9 per cent, compared with 12.9 per cent for the standard catheter (P = NS). The rate of CRS for the ARROWgard was 8.7 per cent, compared with 8.1 per cent for the standard catheter (P = NS). The coating of central venous catheters with silver sulfadiazine and chlorhexidine does not reduce the rate CRI or CRS when compared with standard central venous catheters in patients receiving TPN. PMID- 8712562 TI - Computed tomography: an unreliable indicator of pancreatic trauma. AB - Computed tomography (CT) is currently the modality of choice in evaluating pancreatic injury in patients suffering abdominal trauma who do not require immediate exploration. The purpose of this study was to determine the reliability of initial CT scanning in the detection of pancreatic trauma. A retrospective review was performed of all patients admitted to two Level 1 trauma centers over a 10-year period. Those patients identified with pancreatic injury who underwent initial evaluation with CT scanning were reviewed for clinical course and comparison of CT results with findings at laparotomy. Seventy-two patients of 16,188 admissions (0.4%) were identified with pancreatic injury. Mechanism of injury was blunt in 27 (37%), gunshot wound in 32 (45%), and stab wound in 13 (18%). There were 18 (25%) grade I, 32 (45%) grade II, 16 (22%) grade III, and 5 (7%) grade IV pancreatic injuries. Seventeen of the 72 patients with pancreatic injury underwent initial abdominal CT. The pancreas was normal on CT in 9 and of these, 8 underwent exploration, most commonly secondary to splenic injury. Three were found to have grade I pancreatic injury, two grade II, and three grade III, which required distal pancreatectomy. The pancreas was abnormal on CT in eight patients, and of these three underwent exploration. One patient had an injury upgraded from II on CT to III at exploration and underwent distal pancreatectomy. The mean pancreatic injury by CT was 0.45 versus 2.0 on exploration (P < 0.001). Injury to the pancreas following blunt trauma is rare. Computed tomography will often miss or underestimate pancreatic injuries that require operative treatment, and normal findings on initial scan should not be relied upon to exclude significant pancreatic trauma. PMID- 8712563 TI - The arteriovenous fistula for hemodialysis access: gold standard or archaic relic? AB - A five year retrospective review was undertaken to evaluate the patency rates of arteriovenous fistulae (AVF) in patients with end stage renal disease. From July 1989 through June 1994, 150 fistulae were created in the wrists of 144 patients. Thirty-four percent of the patients had diabetes mellitus. Patient death or irreparable fistulae were considered end points in the study. Patency rates were calculated by the Kaplan-Meier Actuarial Analysis. An analysis to assess the impact of the demographic characteristics, underlying renal disease, and effect of revisions on patency rates was calculated. The results demonstrate a high initial failure rate (less than 1 month) of 13 per cent in the entire cohort undergoing fistulae replacement. The 1 and 5-year patency rates were 56 per cent and 30 per cent, respectively. Diabetics had a significantly lower patency rate at 1 and 5 years (42% and 18%) respectively. Others, who had poor patency rates, include patients 70 years old or greater (40% patency at one year). The results suggest that the AVF should not be the first choice of access in elderly diabetics and that these patients would be better served with other modes of access, such as synthetic conduits or permanent indwelling venous catheters. PMID- 8712564 TI - Surgical treatment of metastatic melanoma of the small bowel. AB - Eighteen of 1420 patients with primary cutaneous melanoma presented with symptomatic small bowel metastases and were reviewed to establish the role and efficacy of surgical intervention. The median interval between treatment of the initial skin lesion and detection of the intestinal metastases was 4.4 years (range, 2 months to 15 years). Most patients presented with either anemia, abdominal pain, bowel obstruction, or intussusception. In six patients, small bowel involvement was the first sign of metastatic disease. Seventeen of the 18 patients underwent laparotomy, and all overt metastases were completely excised in 12. Three patients died postoperatively. Fourteen of the 17 patients had satisfactory palliation with complete symptomatic relief. Median survival after resection was 13 months (range, 2 days to 300 months). Median survival of the 12 patients in whom all macroscopic disease was resected was 44.5 months (range, 2 300 months), whereas the median survival in the four with incompletely resected tumors was 4 weeks (range, 2 days-24 weeks). Five of 12 patients who underwent complete resection of small bowel metastases survived more than 6 years, 3 of whom remain well and free of disease at 6, 14, and 25 years. These results justify active surgical intervention in patients with symptomatic small bowel metastatic melanoma, both for relief of symptoms and prolongation of life. PMID- 8712565 TI - Neuralgia after inguinal hernia repair. AB - Severe chronic pain after groin hernia repair is uncommon but potentially debilitating. Fifteen patients with this condition were retrospectively reviewed. All patients had severe pain, which prevented their working or normal activity and was refractory to nonoperative treatment. Essentials of therapy included 1) a preoperative attempt to identify the involved nerve and 2) high ligation and division of the involved nerve identified at exploration. Twelve patients obtained excellent results and were able to return to normal activity with no requirement for analgesia. Understanding of the typical nerve anatomy, as well as the individual variation in nerve anatomy, can help prevent this complication and is essential for correction if the complication does develop. PMID- 8712567 TI - Operations for gastric ulcer: a long-term study. AB - This study assesses the long-term results of operations for benign gastric ulcers. Three-hundred forty-nine patients operated upon between 1950-1979 have been followed over the past 20 years with a mean and median follow-up of 11.8 and 11.1 years. Fifty-five per cent of the patients had a gastric resection without vagotomy; 19.8 per cent had gastric resection with vagotomy; and 20.3 per cent had vagotomy, pyloroplasty, and wedge excision or biopsy of the ulcer. Operations were selected based on the type of ulcer (Types 1-4), whether the surgeon suspected cancer preoperatively, whether the operations was elective or an emergency, and the age and general health of the patient (presence of significant co-morbid disease). Overall mortality was 6.9 per cent, with a mortality for elective operations 3.6 per cent, and for emergency operations of 32.5 per cent. Age and cardiovascular disease were significant factors in operative mortality and morbidity. All operations were equivalent in long-term results. Excellent to good results were obtained in 92 per cent of patients, with an ulcer recurrence rate of 4 per cent. We conclude that vagotomy, pyloroplasty, and wedge excision or biopsy of a benign gastric ulcer is a comparable operation to a more major gastric resection, with or without vagotomy, in the surgical management of gastric ulcer. The addition of vagotomy to gastric resection does not appear to improve long-term results. PMID- 8712566 TI - Low-grade dermal angiosarcoma of the breast following radiotherapy. AB - Dermal angiosarcoma is a rare complication of radiotherapy. It is characterized by an aggressive nature and is distinct from other forms of angiosarcoma in that it does not always present with chronic lymphedema. A few case reports have appeared of cutaneous angiosarcoma arising in women previously treated with breast-conserving treatment and radiation for breast cancer. A review of the literature reveals that these lesions are difficult to diagnose because of their rarity and their highly variable and benign appearance, which may mimic radiation induced changes in the skin. The majority of the literature reports describe high grade lesions; only one case of low-grade angiosarcoma was described. We now report a second case of low-grade angiosarcoma arising 10 years after segmental mastectomy, axillary lymph node dissection, and radiation treatment for infiltrating ductal carcinoma. The prompt diagnosis of dermal angiosarcoma is strongly dependent upon a high index of suspicion. Biopsy should be considered in patients who present with new skin lesions after radiation treatment for breast cancer. PMID- 8712568 TI - Ventral hernia repair with simultaneous panniculectomy. AB - The repair of a ventral hernia in an obese patient presents an interesting clinical challenge. We retrospectively reviewed the charts of 55 patients who, over a 12-year period from 1983 to 1995, concomitantly underwent both ventral herniorrhaphy and panniculectomy or abdominoplasty. In six of 55 patients, the hernia was recurrent. Forty-six patients had primary abdominal wall hernias or diastasis recti. Nineteen of 55 patients had weight greater than 200 lbs. This last subset of patients had a significantly higher incidence of complications, such as seroma, cellulitis, and persistent wound drainage. In our 55 patients, we experienced only two hernia recurrences (3.6%) during an average patient follow up of 53 weeks. From this experience, we believe that simultaneous ventral hernia repair and panniculectomy is a safe and efficacious approach to these two problems so commonly found in the obese patient. Patients with a preoperative weight greater than 200 lbs can be expected to have a greater risk of wound complications. In all cases, the wounds eventually healed with no long-term sequelae. PMID- 8712569 TI - Diagnosis and treatment of iatrogenic spinal accessory nerve injury. AB - Although iatrogenic injury to the spinal accessory nerve in the posterior cervical triangle is a well-described phenomenon, diagnosis can prove difficult and is often incorrect or delayed. We describe a series of six men and three women (mean age 40 years; range, 20 to 52 years) with iatrogenic spinal accessory nerve injuries. Injuries resulted from lymph node biopsies in the posterior cervical triangle in eight patients and posterior foss surgery in one. Eight patients lost the ability to abduct their arm but could still shrug their shoulder, a pattern that resulted in an incorrect initial diagnosis in five patients. The average delay from injury to referral was 8 months. Seven patients underwent nerve exploration an average of 9.7 months after injury. Five had transected nerves that could be repaired; three of these patients required a nerve graft. Pain was greatly relieved in the five patients who had severe pain before surgery, and weakened shoulder abduction improved in four of six patients. Nerve exploration should be considered when the patient's clinical exam does not improve within 3 months of injury. Nerve repair frequently reduces pain and improves shoulder abduction, even 12 months after injury. PMID- 8712570 TI - Unnecessary preoperative investigations: evaluation and cost analysis. AB - In keeping with national efforts to curb escalating health care costs, the necessity of multiple preoperative investigations was evaluated in 60 randomly selected ambulatory surgery patient records. Necessity for testing was assessed on clinical indications, and overall cost was calculated from the rates at both the local Department of Veterans Affairs Medical Center (VAMC) and a community hospital. Two thirds of the investigations were deemed to be inappropriate, with derived unnecessary average cost per patient of $47 and $80 for the VAMC and community hospital, respectively. Potential savings at the VAMC of $11,757.50 for the calendar year could have been realized. Education of staff and housestaff is crucial to changing obsolete practice habits. The quality and safety of care would not be compromised by limiting preoperative investigations to only those with clinical indications. PMID- 8712571 TI - William Beaumont. PMID- 8712572 TI - [Prevalence of HIV infection in Italian drug users attending drug treatment centers in the years 1990-92]. AB - Repeated cross-sectional surveys have been conducted to collect information on the trend of HIV prevalence among drug users attending drug treatment centers in Italy in the years 1990-92. Annual HIV prevalence was 31.5% in 1990, 29.1% in 1991, and 24.6% in 1992, showing a declining trend. HIV prevalence was higher in the North than in the South of the country, and among females than in males. Drug users entering treatment for the first time during the study period were less likely to be infected than old clients. PMID- 8712573 TI - [Health education and the prevention of HIV infection in schools]. AB - This review describes the HIV prevention strategies adopted since 1990 by the Italian Ministry of Health and Ministry of Education, coordinated by the National Health Institute, for use in Italian schools. It sets out reasons for believing that action in schools is essential in containing the spread of the HIV epidemic and presents teaching materials prepared for school use. An analysis is made of the IV national HIV information campaign, in which the Ministry of Health trained 4,000 middle and senior schools principals. The prospects for continuing the work with these 4,000 principals in the V information campaign, are also reported. PMID- 8712574 TI - [Epidemiology and prevention of HCV infection in dialysis centers]. AB - From many epidemiological studies comes out that haemodialysis patients are at high risk of hepatitis C virus (HCV) infection. This review shows the results of several prevalence and incidence studies carried out in dialysis centres. The prevalence of anti-HCV antibodies in haemodialysis patients varies from 20% to 38%. From these surveys comes out that serodiagnostic tests, usually employed in laboratory diagnosis (ELISA and RIBA), may underestimate the true prevalence of HCV infection. The very important risk factors in the haemodialysis units are blood transfusion and the length of time on haemodialysis. Nevertheless, these are not sufficient to explain completely the high prevalence of subjects HCV positive among the dialysis patients. Contamination of environmental surfaces, caused by a non correct application of the asepsis and disinfection techniques, could be, according to several authors, the more important cause of this infection among the dialysis patients. PMID- 8712575 TI - [Poliomyelitis surveillance: seroepidemiologic study on a Piedmont population sample]. AB - In order to assess the immunity/receptivity towards the three poliovirus strains, a sample representative of the sex and age composition of the resident population in Piedmont at census 1981, was examined. For each subject data were collected in order to identify the population from social-ambient point of view. For the evaluation of antibodie's title to poliovirus 1, 2, 3, the serums were analyzed with the neutralization method using the microtitolation plates and epithelial larynx cancer cells (HEp-2), like revealing system. The samples with > or = 2 title were considered positives for specific antibodies. To determine the relation between presence/absence of specific poliovirus antibodies and the other variables, a multiple logistic regression was fitted and the odds ratio was calculated. The results of our study show an immunity in all age groups, underlying a herd immunity condition. Furthermore the incomplete antibodies response to the three poliovirus strains seems to be influenced by age only. PMID- 8712576 TI - Gluten-sensitive enteropathy. AB - Peptides originating from wheat gluten during digestion in the alimentary tract are known to cause primary intolerances in genetically-predisposed individuals. Other cereals such as rye, barley and probably oats are also toxic for coeliac patients, whereas rice and maize are considered to be non-toxic. The mechanism by which prolamine-derived peptides produce jejunal lesions is not fully understood. Most investigators favour a dysregulated immune response to gliadins as the underlying abnormality in coeliac disease, but according to other authors a non immuno-mediated cytotoxic activity of gliadin peptides on the small intestine seems to be the primary cause of intestinal mucosal damage in coeliac patients. This paper is a critical appraisal of current theories on the pathogenic mechanism underlying this disease. Moreover, many in vitro systems needed to investigate the cereal toxicity are described. PMID- 8712577 TI - [Ras gene analysis in mammary tumors of dogs by means of PCR-SSCP and direct genomic analysis]. AB - The oncogenic capacities of RAS family genes (Ha-ras, Ki-ras, and N-ras) are usually activated by point mutations in the conserved regions (codons 12, 13, and 61), resulting in single amino acid substitution in the specific proteins (p21). In order to verify the involvement of RAS genes in dog mammary tumors we analyzed the genomic DNA from 20 mammary tumors of dog by means of the Polymerase Chain Reaction-Single Strand Conformation Polymorphism (PCR-SSCP) method and the direct genomic sequencing. The absence of point mutations in the "hot spots" of RAS genes suggests a lack or a low frequency of such a pattern of RAS genes activation in dog mammary tumors. The results are also in agreement to what reported in human mammary tumors. However, the presence of genetic alterations in other functional areas of the RAS genes or other mechanisms of activations cannot be ruled out. PMID- 8712578 TI - [Allergy to house dust mite: a public health problem]. AB - Mites have been recognized as the most important allergen in the house dust. This review deals with morphology, ecology and biology of house dust mites including factors affecting the distribution, abundance and seasonality of the species. Clinical, diagnostic and therapeutical aspects of the allergy to domestic mites are also reviewed. Finally, preventive and control measures against house dust mites are suggested. PMID- 8712579 TI - Clinical pharmacokinetics of cumulative very high dose of cisplatin in chemotherapy resistant solid tumors. AB - Cisplatin was administered to seven patients with advanced cancer in divided doses of 40 mg/m2 body surface daily for 5 consecutive days. The pharmacokinetics of total Pt was studied on the days 1, 3 and 5 of infusion. The renal function was assessed through the parameters usually applied in the clinical practice (serum creatinine level, creatinine clearance, urinary volume). Pt pharmacokinetics and the renal function did not show modifications outside the normal range. However, on day 5 of treatment patients showed increased alpha-half life and AUC of plasma Pt, as well as decreased Pt total body clearance and Pt renal clearance, associated to a significant (although still within normal range) increase in serum creatinine and a decrease in urinary volume. Moreover, a correlation between Pt pharmacokinetics and renal parameters (measured as the difference between the values of days 1 and 5 of treatment) was also found: the increase in creatinine was directly related to a decrease in Pt renal clearance and inversely related to Pt peak level in urine, while the latter was inversely related to a reduction of Pt renal clearance. It was concluded that very high doses of cisplatin are well tolerated in patients, although some parameters might suggest early impairment of the renal function. PMID- 8712580 TI - Analytical problems in mercury analysis of seafood. AB - It is generally accepted that seafood represents one of the major sources of mercury to man. In this work two interlaboratory proficiency tests are described for the analysis of mercury in seafood. Thirty-seven public control and food industries laboratories participated in the first test, while 29 participants were included in the second one. Moreover, in order to clarify whether sampling of different edible muscle tissues of the same fish could affect the analytical results, the top, the central and the bottom portion of 28 fishes were examined. The different portions of fish showed no significant difference in mercury concentrations. Two different wet digestion methods (microwave oven and reflux in quartz vessels) were also tested in the case of 11 fishes. A systematic difference was observed between the two sets of results obtained with these digestion methods. PMID- 8712581 TI - Daily surface ozone maximum concentrations at Taliarte, Canary Islands. AB - Two different methods for using air quality data to estimate daily maximum concentrations along a year of surface ozone concentration in a new measurement station in Canary Islands are described. It is usually assumed that sequences of pollutant concentration show stationary behaviour. In this paper, it is shown that the application of such procedure could be useful to estimate probabilities of maximum concentration of surface ozone. Nevertheless, this method does not take into account the time structure of the original data, so that some caution should be taken into account. In this sense, an alternative procedure, assuming nonstationary behaviour of data, is developed to estimate empirically the probability distributional properties of the original series. Its application yields a substancial improve of the explanation of underlying series structure. PMID- 8712582 TI - [Very low frequency electric and magnetic fields and the immune system]. AB - Recent residential and occupational epidemiological studies indicate a statistical association between 50-60 Hz magnetic field exposure and the risk of developing some kinds of tumors. Several experimental researches have been carried out in vitro and in vivo to verify the possibility that some cell functions may be influenced by ELF (Extremely Low Frequencies: 0-300 Hz) electric and magnetic fields. Such researchers are very important to assess if the statistical association indicated by the epidemiological studies is actually due to a cause-effect relationship between ELF electric and magnetic fields and carcinogenesis. In this review we describe the present state of the experimental research, focusing our attention on the effects of ELF fields on the immune system. We also describe some theoretical researches whose aim is to identify possible mechanisms of interaction between ELF fields and biological systems which may provide biological plausibility to the observed effects. PMID- 8712583 TI - [Scientific literature: bibliometric and bibliographic indicators as integrative criteria for an objective evaluation of research activity]. AB - An objective evaluation of a scientific activity is required to reduce partialities and mistakes, thus allowing a more reliable judgement. Therefore, the role of scientific literature, as the most direct expression of research activity, is increasing; for a better evaluation, bibliometric and bibliographic indicators have been developed. Bibliometric indicators are based upon citation, which is a meter of the use the international scientific community makes of research results, once published. Besides the number of publications, the most important bibliometric indicators are the number of citations, the impact factor, the immediacy index, the half-life index. Bibliographic indicators are the reference of the journal or the paper by a given author, in important bibliographies of international influence. Being the evaluation by means of these indicators extremely complex, there is a need for a homogeneous regulation of the activity, at an institutional level. The presence of experts in scientometrics is also advisable, since they can provide evaluation profiles on demand, with the support of modern documentation centres. In sum, after describing bibliometric and bibliographic indicators, emphasis is put on their utility and the role they play in support of experts' judgement for a more precise evaluation of a scientific activity. PMID- 8712584 TI - [Etiopathogenesis and physiopathology of hemorrhoidal disease]. AB - Many possible causes have been proposed to explain the pathogenesis of symptomatic hemorrhoids: constipation and abnormal bowel habits are commonly blamed despite largely contrary evidence. The most consistently demonstrated physiological abnormality is an increased maximum resting anal pressure. The pathophysiologic theory, made popular by Thomson's studies, in which the anal cushions break their elastic support and become more sensitive to the increase of pressure from straining and to trauma from hard stool, with a progressive downward displacement, is currently the best accepted. The intimate mechanism of functioning and control of the arteriovenous anastomoses remains unclear as does the existence of an individual susceptibility in which a part is played by socioeconomic, cultural and psychological factors. PMID- 8712585 TI - [Clinical manifestations and diagnosis of hemorrhoids]. AB - Haemorroidal disease has often typical presentation with rectal bleeding, anal prolapse and pain. Practitioners and patients usually underestimate the symptoms; in adults and older people the same symptoms may suggest neoplastic disease, which may be synchronous. Proctoscopy is the main diagnostic test for staging; endoscopic examination with biopsy is helpful to differentiate neoplastic, granulomatous and inflammatory anorectal lesions. In our mind, a complete study of anorectal function and morphology is necessary for a correct diagnosis. PMID- 8712586 TI - [Anorectal manometry in hemorrhoidal disease]. AB - Manometry is an important approach to anorectal function. In the haemorrhoids the evaluation of the anal resting tone, of the squeeze and of the sphincterial length is very useful to realize the best surgical treatment. Anorectal manometry is neither invasive nor expensive; it can be ready repeated and the results are reproducible. This examination must be considerated as essential part of the diagnostic routine of the anorectal diseases. PMID- 8712587 TI - [Conservative therapy of hemorrhoids]. AB - Conservative therapy of hemorrhoids can be curative at stage I and II of disease; further it rappresent a valid preliminary management for a better result of more aggressive treatments, be they surgical or not. Beside local and general drugs other aspects play an important role in conservative therapy of hemorrhoids: adequate local hygiene, correct diet, education to a physiological evacuation and last but not least stool frequency regulation. PMID- 8712588 TI - [Sclerotherapy and elastic ligation of hemorrhoids]. AB - In the treatment of haemorrhoids all procedures may have good results, if indications are correct. Injection treatment has the best indication for I and II degree bleeding piles, the rubber band ligation for II and III degree piles; this procedure is very effective when associated to cryosurgery. All operations for haemorrhoids must be avoided in Crohn disease. PMID- 8712589 TI - [Infrared photocoagulation, cryosurgery and laser surgery in hemorrhoidal disease]. AB - The non invasive treatment of hemorrhoidal disease has the best indications for I and II degree symptomatic piles and only in few cases of III and IV degree. Authors report their practice with infra-red-coagulation, cryosurgery and laser surgery, discussing indications, controindications and describing in detail their technique. About laser surgery they emphasize that this procedure is a formal hemorrhoidectomy, and laser acts as a scissor or a blaude. PMID- 8712590 TI - [Hemorrhoidal thrombosis and perianal hematoma: diagnosis and treatment]. AB - Thrombosed haemorrhoids and anal haematomas are very usual in patients with haemorrhoids. Conservative treatment and surgery are effective by the features and time of presentation. Authors refer about pathological and clinical findings and discuss the treatment. PMID- 8712591 TI - [Closed hemorrhoidectomy]. AB - In the closed hemorrhoidectomy wounds are completely sutured. Ferguson and Whitehead operations, the latter modified in order to avoid mucosal ectropion and stenosis, are the most performed closed hemorrhoidectomies. 15% Ferguson operations, 10% Whitehead-Rand operations, and 75% Milligan and Morgan operation were performed among 400 hemorrhoidectomies. Author discusses indications for closed hemorrhoidectomy; techniques are described, complications and results reported. PMID- 8712592 TI - [Hemorrhoidectomy: Milligan and Morgan technique]. AB - Milligan and Morgan and Arnous anoplasty procedures are described; their indications come from pathological features and associate pathology. Left internal sphincterotomy and posterior sphincterotomy and anoplasty are described and discussed on the basis of the Author experience about 523 haemorrhoidectomies. Internal sphincterotomy, when indicated, avoids pain, urinary retention, fissure and stenosis. Early and late complications are evaluated for both the procedures. PMID- 8712593 TI - [Treatment of hemorrhoidal peduncles with diathermy probe]. AB - Diatermy hemorrhoidectomy, without suture of hemorrhoidal pedicles, is a variant of the classical Milligan and Morgan operation. This procedure, if correctly performed, reduces postoperative pain and complications; results are good. Great care is reserved to internal sphincter; it must not be involved in coagulation. Evaluation concerning 100 personal cases are referred. PMID- 8712594 TI - [Submucosal reconstructive hemorrhoidectomy (A.G. Parks operation)]. AB - Indications and procedure of submucosal reconstructive hemorrhoidectomy (A.G. Parks hemorrhoidectomy) are described and discussed. This procedure is more difficult and time consuming than other techniques, but results are better in high degree hemorrhoids. Authors consider complications and sequelae on 278 patients operated in their Department. PMID- 8712595 TI - [When to combine internal sphincterotomy with hemorrhoidectomy]. AB - 396 patients, treated with hemorrhoidectomy (278 Parks, 118 Milligan-Morgan) were submitted to preoperative manometry, which, in 158 patients (39.9%), revealed high anal resting pressure (TBS), with or without an associated anal fissure; this group was submitted to a regulated lateral sphincterotomy (SILR) in association with the haemorroidectomy. The manometric follow-up, two mounts after the operations showed similar TBS values in both groups of patients. The authors conclude that normal values of TBS after Hemorroidectomy can be reached only by associating sphincterotomy in manometrically selected cases. PMID- 8712596 TI - [Complications of the surgical treatment of hemorrhoids and its therapy]. AB - Hemorrhoidal disease is a common problem in a proctological ambulatory. Surgery is the best therapy for fourth degree hemorrhoids and the complication rate is 10 20%: postoperative urinary retention etiology is unknown but it may be caused by dysfunction of bladder muscles in response to pain and by an excessive perioperative somministration of fluids; delayed hemorrhage (i.e., 7-10 days postoperative) needs an inpatient care and the treatment ranges from bedside and packing to hemorrhoid pedicle suture ligation in the operating room. Anal stenosis is most commonly a result of a prior improper hemorrhoidectomy: it may be mild, moderate or severe; V-Y and C-anoplasty are the best therapy, also for ectropion. Rubber band Ligation and Sclerotherapy are the most common treatment of internal hemorrhoids: external hemorrhoid trombosis and delayed hemorrage are frequent complications. PMID- 8712597 TI - Management of descending aortic dissection. AB - The optimal timing of surgery with Stanford type B aortic dissection remains controversal. In acute-phase cases, surgical mortality is so high that medical treatment is preferable unless there are major complications. To guide the choice of medical versus surgical therapy we use survival analysis in patients with acute uncomplicated/complicated and chronic uncomplicated/complicated descending aortic dissection. Between 1992 and 1993 49 patients were diagnosed with Stanford type B aortic dissection. Emergent surgery was performed in 4 patients for rupture or impending rupture, elective surgery was done in 12 patients. The remaining 33 patients were treated medically. Our results support the continued use of medical management as the primary treatment for uncomplicated acute aortic dissection, with surgical therapy being reserved for those patients with complications such as rupture, expansion, continuing pain or ischemia of distal vascular beds. PMID- 8712598 TI - [Surgical treatment of differentiated thyroid carcinomas: results in 440 cases operated between 1970 and 1994]. AB - The authors report their experience and surgical treatment of 440 patients with differentiated carcinoma of the thyroid (DCT) from 1970 to 1990 in the III Surgical Dept. at the University of Rome "La Sapienza". Considering 305 cases of papillary carcinoma, 103 Total Thyroidectomy (TT), 118 TT with Functional Regional Lymphadenectomy (FRL), 32 TT with Node-Picking, 51 TT with curative FRL and one TT with Radical Neck dissection were performed. In 135 cases of follicular carcinoma TT associated with Node-picking were performed on two patients, TT associated with FRL on 8 patients and TT associated with RND on 2 patients. Overall survival to 10 years was 90%; 98% for N- papillary carcinoma and 66% for N+ follicular carcinoma. There were no permanent recurrence of disease. Out of the patients who did not undergo cervical lymphadenectomy 30% developed temporary hypocalcemia; 53% (mediastinal lymphadenectomy) to 87% (FRL) of the patients submitted to lymphadenectomy developed temporary hypocalcemia with permanent hypoparathyroidism in 0.9%-23% of the cases. On the base of what has been observed by Pasieka and Grant, the Authors believe TT as the absolute solution for patients with DCT because it allows the removal of both the original tumor and the possible neoplasms in the residual gland tissue, and enables early identification of other metastases which can be treated efficaciously with I131. PMID- 8712599 TI - [Endoscopic & histopathologic features of esophageal ulcers. Considerations in 13 cases]. AB - Chronic, severe reflux esophagitis can be complicated with an esophageal ulcer (EU), whose histomorphological characteristics change sensibly. Between 1983 and 1989, 18 EU were endoscopically identified in 13 (6%) out of 217 patients with chronic gastroesophageal reflux (GER). 2 groups were defined: EU and esophagitis, without columnar-lined esophagus and EU with localized columnar metaplasia or Barrett's esophagus. In the first, 4 Wolf's ulcers (WU) were present; in the second, there were 8 Barrett's ulcers (BU), 2 Savary's ulcers (SU) and 4 ulcers with intermediary characteristics (IU). The 24 h esophageal Ph monitoring demonstrated the reflux pathogenesis in all the patients (57%: alkaline reflux). The histomorphological findings of the EU could support the hypotesis that their variety is related to the different stage of the reflux disease. The EU, started as a peptic lesion on a stratified squamous mucosa (WU), due to an acid, alkaline or mixed reflux, would macroscopically turn into a BU, through intermediary entities (SU, IU), with a progressive extension of columnar metaplasia to the adjacent epithelium and walls. It's not possible to exclude the primitive development of the BU in a Barrett's mucosa, because of an alkaline reflux or because of a low mucosa's resistance (specialized columnar epithelium) to a prolonged acid exposure. PMID- 8712600 TI - [Total gastrectomy for gastric neoplasms]. AB - In spite of the decreasing incidence of gastric carcinoma, as it is reported in recent reports from the U.S.A., total gastrectomy and its surgical indications results and complications, focus the interest of surgeons. We analize 61 cases of total gastrectomy for carcinoma, treated in the years 1982-1992. Perioperative mortality and long term survival appear highly satisfactory, mainly if one considers that the site and extension of the neoplasms treated would have severely impaired the possibilities of cure or long term survival with surgery of lesser momentum. We believe that nowaday indications for total gastrectomy can be widened in the hope of improving results of gastric cancer surgery with no or little additional risk. PMID- 8712601 TI - [Crohn's disease in the elderly]. AB - Primary Crohn's disease in the elderly is a not frequent pathology, which however have distinguished from the reheightening or relapse of an inflammatory illness risen up in juvenile age. The Authors compare the personal experience (6 cases on 120 patients operated for Crohn's disease) with the data of the literature. The more frequent localization stays the ileo-colic one, also if in the elderly present better impact the cases of Crohn's colitis (pancolitis or left colitis), with frequent and- perianal implications. The symptomatology is not pathognomonic and above all in the cases of colitis sets problems of differential diagnosis with other pathologies, among which the diverticular disease, that on the other hand, can also coexist with the inflammatory illness. A pharmacological treatment is desiderable, but frequently doesn't result effective. The surgical conservative therapy is not always possible and could be necessary resort to maiming interventions, like massive ileal resections or a total proctocolectomy. From the prognostic point of view, the course appears from the beginning or very favorable (with low index of relapse) or tumultuous (with acute manifestations, which require an emergency surgery, wighted by a more elevated rates of mortality and morbidity. PMID- 8712602 TI - [Chronic gallbladder disease & its surgical therapy, personal experience]. AB - The chronic alitiasic cholecystopathy represents an unusual trmorphological disease not well defined in literature; it is characterized by a chronic alteration of gallbladder without gallstones. The authors present a retrospective study on a group of 21 patients having undergone cholecistectomy from January 1992 to January 1995. The choice for the surgical treatment was made on the basis of blind gallbladder after i.v. cholangiography, and cholecystography and/or tickning of the gallbladder wall performed through ultrasonography, in patients with hystory of biliary pains not responding to the pharmacological therapy. The removed gallbladder has been histologically examined, observing the absence of gallstone and the presence of chronic inflammation of the gallbladder wall. The c.a.c. is a pathology whose etiopatogenesis is yet discussed, some authors assert in fact that on its basis there are functional symptoms. Although in the past these kinds of disease were considered functional disorders this study revealed that in some cases there are organic problems and that the surgical treatment is necessary. PMID- 8712603 TI - [Biliary calculi in liver cirrhosis. Clinical, epidemiologic & therapeutic aspects]. AB - BACKGROUND: Incidence of cholelithiasis in cirrhotic patients is twice of threefold that in non cirrhotic one. Cholelithiasis is often asintomatic, however, when complications occur, emergency surgery is burden by high rates of morbidity and mortality. Purpose of the current study is to state the clinical features and to assess the outcome of elective surgery in these patients. METHODS: Sixtyseven consecutive patients with liver cirrhosis (Group A) who underwent elective surgery for biliary stones were retrospectively reviewed. Preoperative clinical features, surgical procedure, postoperative morbidity and morality were recorded and compared with those of the 3291 non cirrhotic patients (Group B) observed in the same period and submitted to elective surgery for biliary stones. RESULTS: Stone involvement of the biliary tract (A vs B:31.3% vs 20.5%), cholangitis (A vs B:23.8% vs 13.0%) and acute biliary pancreatitis (A vs B:4.4% vs 0.7%) were found to be more frequent among cirrhotic patients. In Group A postoperative morbidity and mortality in Child-Pugh A and B were found to be not statistically different from those of Group B patients. CONCLUSION: Authors conclude that definitive biliary surgery may be considered as a successful and safe indication in the treatment of cirrhotic patients in Child-Pugh A and B grade. Conservative approaches are considered as more suitable in Child-Pugh C patients, and definitive elective procedures should be considered for these patients only when an improvement of their liver function can be achieved. PMID- 8712604 TI - [Injury of the popliteal artery & the popliteal branches: analysis of 31 consecutive cases]. AB - In complex traumas of lower limbs it is fundamental the reducing of the time of ischemia to reduce the number of failures in repairing surgery and the percentage of the demolishing of necessity surgery. The experience of the authors is based on 31 complex traumas of lower limbs with distal vascular injuries to Hunter's canal, which were associated to bone, nervous and muscular lesions. In total we are treated 37 vascular injuries by interposition of venous autografts in 37 cases, lateral pacth in one case ant suture T-T in two cases. The protection of the reconstructed vases, in case of a concomitant loss of substance, was entrused to microvascular flaps for 7 times latissimus dorsi, iliac crest and fascio cutaneous for 5 times posterior reversed with distal baset and for three times antero-lateral, which in three cases of them needed successively a dermoepidermic grafts. The bone injuries, which were treated by external fixation, intramedullary nailing, plate and screws, just in two cases became worse in pseudoarthroses and just in one in osteomyelitis. The reparation of nervous injuries had bad results just in two cases. The percentage of saving of the limb it was about 83.9%. PMID- 8712605 TI - [Inguinal hernia repair in the elderly: modified Shouldice procedure, 10 years of experience]. AB - Authors relate their experience of 10 years period (1984-1994) with Shouldice technique for inguinal hernia repair in the elderly. This technique, in the Author's clinical practice, replaced the Bassini operation. The personal series is costituted of 190 patients underwent to inguinal repair during a 10 years period from 1984 to 1994 with recurrence rate of 1.1% (2 pt.). Authors consider Shouldice operation to be most valid from the methods of direct suture, with lower recurrence range. PMID- 8712606 TI - [Simultaneous multiregional embolization from a cardiac myxoma. Case report]. AB - Atrial myxoma is the most frequent cardiac tumor. Distal embolization occurs in about one third of the patients with this nosological entity; a simultaneous embolization to three different arterial districts, however, is a rare event. Of particular interest is also, in our opinion, the evolution of the clinical picture and the surgical treatment performed. PMID- 8712607 TI - [Afferent loop obstruction after total gastrectomy presenting as acute pancreatitis (clinical contribution to the etiopathogenesis)]. AB - Afferent loop obstruction presenting as acute pancreatitis is an uncommon pathology encountred after total gastrectomy or gastric resection. The authors refer about a case of afferent loop syndrom presenting as acute pancreatitis, and conclude that, since this disease is very rare, aggressive and variable, its diagnosis and therapy is still controversal. PMID- 8712608 TI - [Renal angiomyolipoma causing an acute abdomen]. AB - Herewith referred a case of renal angiomyolipoma relieved on a patient of 62. The new involvement has been found following an evidence of a keen syntomathology related by a steep abdominal ache centred above all on the right hypocondrium followed by a general light ache. The ecography and the "TAC" on the bely have showed a new enlarged involvement, centred on the hypocondrium and on the right oval side of a diameter of nearly 35 cm, that moved anteriorly the right renal, with an involvement of a perirenal hematoma. The surgery has ended with a nephrectomy with the drop of the involvement on the renal tissue and the emorragical enlargement which has allowed a local nephrectomy and therefore to resume the organon as a new growth not dangerous. The histological diagnosis separated on the renal carcinoma han been possible with certainty only examining cruently the area after a tumorectomia, but the general clinical course and the evidence of the angioTAC had been realized towards the exact diagnosis. PMID- 8712609 TI - [Functional evaluation of an experimental model of cutaneous microcirculation and pO2 after surgical excision: traditional method vs laser CO2]. AB - Within an experimental study project about the interaction between Laser and biological tissues, the authors utilized a Laser Doppler Flowmeter device and a Polarographic method to study the cutaneous microcirculation and the tissue pO2. Under standardized conditions, the experimental model considered the microcirculation measurement and the ptiO2 before and after the exsection of a cutaneous lozenge on the animals back realized with traditional surgical methods (scalpel and electrotome) or with a last generation CO2 Laser in continuous and in pulsed mode. The Laser Doppler Flowmeter showed that the largest perturbation of the microcirculatory flux occurred after the exsection realized with the scalpel and the electrotome. The CO2 Laser utilized in continuous mode showed an influence slightly lower on the microcirculation while in pulsed mode the perturbation was absent. The ptiO2 measurement showed similar results emphasizing that the CO2 surgical Laser technique is less invasive than the traditional. To sum up, these functional methods allowed a careful microcirculation an ptiO2 evaluation and provided useful information about vasal and metabolic alterations, showing that the CO2 surgical Laser utilized in pulsed mode appears to be the less harmful surgical technique for the tissues surrounding the exsection site. PMID- 8712610 TI - [A case of traditional appendectomy changed to laparoscopic surgery]. AB - The utility of appendectomy performed with laparoscopic procedure is at the present accepted by many surgeons, especially when there is uncertain diagnosis of appendicitis or when an ectopic position is suspected. The authors describe the case report of a 17 years old woman with a diagnosis of subacute appendicitis, in which the unsuccessful finding of the appendix made necessary conversion of traditional laparotomy approach to laparoscopic technique. This choice avoided the execution of an enlargement of laparotomy access, and allowed an easy finding of appendix. Aesthetic results at a distance are very satisfying. PMID- 8712611 TI - [Ambulatory surgery: current status and prospects]. AB - Ambulatory Surgery (AS) was born some 20 years ago in USA pushed by economic requirements and rapidly spread over the European countries as one of the main factors of progress in health care. The Authors, after evaluating the actual situation of AS in the international literature, report their personal experience from January 1991 to December 1994. The series includes 810 surgical operations performed as outpatients procedures with immediate discharge. The following types of anaesthesia were used: local infiltration (86.6%), monolateral ultraselective spinal (10%), blended or general (3.4%). Hernias of the abdominal wall, varicose veins and anorectal diseases were the more frequent pathologies operated on. Results of surgery are satisfactory supporting the advantage of AS such as the absence of complications due to anaesthesia and hospital stay, the better relationship between patient and surgeon, the short return to working activities. AS proves its value on the clinical and socio-economic grounds provided that a well organised program and careful selection of patients are adopted. No extemporary organisation are advisable. PMID- 8712612 TI - Risk factors for surgical wound infections in clean surgery: a multicenter study. Italian PRINOS Study Group. AB - OBJECTIVE: To investigate potential risk factors for surgical wound infections in clean surgery. STUDY DESIGN: Multicenter prospective study in eight general surgical wards and one thoracic surgical ward. METHODS: All patients undergoing clean operations in the nine wards were included in the study. The following data were collected for each study participant: demographic characteristics, underlying disease, predisposing factors, type of surgery, length of operation, preoperative shaving, type and duration of surgical drains, and length of hospital stay. After surgery, patients were visited daily by an intern surgeon in order to detect infections. Infections occurring after discharge were not actively surveyed. A multiple logistic regression was performed to obtain an adjusted estimate of the odds ratios and to identify which factors were independently associated with surgical infection. RESULTS: 2,262 patients were included in the study: eighty three patients (3.7%) developed a surgical infection. The highest infection rates were observed following vascular surgery, thoracic surgery and herniorraphy. Seven factors appeared to be significantly and independently associated with an increased risk of SWIs in a logistic regression model; age greater than 85 years, obesity, patients undergoing high risk operations or more than one operation during stay, length of operation greater than 120, preoperative shaving, open surgical drains lasting more than three days. CONCLUSIONS: The high incidence of infection observed in our study population indicates potential problems in quality of care. In order to prevent SWIs in patients undergoing clean operations, more efforts should be placed on the prevention of the risk associated with two modifiable risk factors, that is preoperative shaving and use of open surgical drains. PMID- 8712613 TI - Traumatic rupture of the descending thoracic aorta. AB - Timing and tactics in the repair of the traumatic ruptured thoracic aorta are matter of controversy ever since. The unmeasurable risk of a consecutive rupture favours a primary repair, concomitant injuries, however, a delayed repair. In single injuries of the thoracic aorta the clamp/repair procedure within 24 hours generates acceptable results with an overall mortality of 4 to 8% and a risk of ischemic myelopathy of 8 to 10%. Delayed repair reduces these figures not at all. Extracorporal circulation produces rather worse results in contrast to clamp/repair procedures. In cases of severe concomitant injuries, e.g. brain damage, hemorrhage and open fractures a delayed repair after cardiopulmonary reconstitution is required. In cases with posttraumatic pulmonary insufficiency the risk of surgical procedure itself is much higher than the risk of a second rupture of the traumatized thoracic aorta. We demonstrate our tactics in the repair of traumatic rupture of the descending aorta displaying clinical operated 1992/93. PMID- 8712615 TI - [Indications of intraoperative cholangiography in laparoscopic cholecystectomy]. AB - After having related about the methods of pre and intraoperative investigation of the biliary tract in video-laparoscopic biliary surgery, the Authors make a list of the advantages and describe the video-laparoscopic methods of intraoperative cholangiography. Moreover the authors review the international literature about the subject and relate the personal statistics and opinions. PMID- 8712614 TI - [B-cell lymphoma (MALToma) of the stomach and Helicobacter pylori infection]. AB - The authors, after illustrating physiopathologic aspects of gastric MALTomas, examine the H.P. infections and the possible relations between this kind of bacterium and gastric MALTomas. They hypothesize that H.P. infection represents an important predisposition to gastric lymphoma, due to anatomopathological modifications over gastric mucosa. They conclude that a decreasing of gastric lymphoma could be correlatable an opportune eradication of H.P. despite of at moment it's impossible hypothesize a marked decreasing of lymphoma by an eradication of H.P. PMID- 8712616 TI - [Follow up in carcinoma of the large intestine]. AB - A valid program of follow-up has always been a crucial point in the overall therapy of the colon-cancer. In this retrospective study, the authors have used as specimen 74 patients put under observation between the years 1987 and 1992. The patient have been followed throughout the diagnostic period with various methods. It has been the will of the authors, who have presented their protocol of reference, to put under comparison the various controlling methods in order to visualize their reliability, specificity and the indication of each one of them. The CEA is the most sensible haemanalysis for lifting the doubt of recidivation. As for the TAC and ultrasound it has been reserved the job of formulating a correct diagnosis; the results of both diagnostics through imagery have been more or less the same. However, the ultrasound examination have shown more false positives than the TAC. The research of the blood occult in the stool is a rapid and economic detection in the case of intramural recidivations, even if we cannot disregard the share of false positives. A high specificity for the study of intramural recidivations has been offered by the endoscopic scan particularly when associated by a brushing and biopsy. PMID- 8712617 TI - [Latero-lateral portocaval anastomosis in the surgical treatment of portal hypertension: evaluation of a mean follow up of 14 years]. AB - After the enthusiasm given by the new treatment techniques of complicated portal hypertension (sclerotherapy, TIPS and liver transplantation), various authors in the literature are considering the role of derivative surgery in the treatment of this affection, due to their consolidated experiences and with a long term follow up. The authors analyse their own long experience, regarding 104 side-to-side porto-caval shunt. It has been done a complete follow-up, of 71 patients, for a period lasting from 4 to 20 years from the operation. Intraoperative mortality was 5,7%, global morbidity was 28%. Haemorrhagic recurrence from rupture of oesophageal varices was found in 2 cases (1 soon after and the other 3 months from the operation). 5 years survival was depending from Child score (96% Child A, 66,4 Child B, 25% Child C) and from the ethology of cirrhosis (70% for alcoholic cirrhosis and 80% for post-hepatitis cirrhosis). The survival was anyhow mainly connected with the persistence of alcohol abuse. The EPS appeared or became worse after the operation in 12 patients out of 71 checked (16.7%). It has never made a patient enable to lead an ordinary life. Due to the results of their own experience, the authors underline the efficacy of side-to-side porto caval shunt in the prevention of the haemorrhagic recurrence from oesophageal rupture. In selected patients, Child A and B, not responding to the endoscopic treatment and in elective operations, the mortality was very well contained with a good prevention even after many years and a very small occurrence of EPS. For all those reasons, the side-to-side porto-caval shunt still keeps its validity also towards the selective shunts that are much more difficult technically and that can't be done on ascitic patients. PMID- 8712618 TI - A study of some international medical journals aiming to elicit stylistic guidelines for writing publishable papers. AB - OBJECTIVE: To provide some indications and guidelines on style and presentation for those aiming to publish in selected international journals. SETTING: University library in the Istituto di Clinica Chirurgica, Bari University Hospital. METHODS: A single topic, portal hypertension, was chosen to ensure a certain homogenity for the purposes of comparison, and the three most recent articles on this topic were identified in six international journals (three published in England and three in the USA; three in the medical and three in the surgical field) and studied from the points of view of style and presentation. RESULTS: Strong similarities were found among the 18 selected articles. In the interests of brevity, all were characterized by compact statements, qualified by lists of examples, and strings of adjectival nouns. In all, the Introduction and Conclusions sections were relatively short, the main body of the article being reserved to the Patients and Methods, Results and Discussion/Comments sections. Apart from very slight spelling or grammar differences characterizing English or American usage, all the articles could, with slight alterations in format in some cases, equally well have been published in any other of the journals studied. CONCLUSIONS: Brevity, even at the expense of elegance, and a straightforward, unambiguous, to some extent standardized, approach to the presentation of a scientific study, are essentials for the acceptance of an article for publication in the selected temational journals. PMID- 8712619 TI - [Role of ultrasonography in abdominal surgical emergencies. Our experience]. AB - OBJECTIVE: To assess the sensitivity, specificity, and predictive value of ultrasonography in surgical patients in abdominal emergency in regard to the indication for immediate operation, delayed abdominal exploration, or conservative treatment. DESIGN: A retrospective study was conducted after consecutive sampling of 98 patients in a control trial. SETTING: The study was conducted at the University Hospital of Catania (Italy), which serves as a general community hospital. PATIENTS: All patients with acute upper abdominal pain or blunt abdominal trauma were eligible for the study. A total of 110 patients were observed from 1990 to 1994, after plain X-ray of the abdomen in 12 patient was diagnosed visceral perforation and they went for immediate operation. The remaining 98 patients were divided into two groups: acute upper abdominal pain (56 patients) and blunt abdominal trauma (42 patients). INTERVENTION: Ultrasonography in the emergency department. MAIN OUTCOME MEASURES: Conservative or operative treatment based on ultrasonographic and clinical findings. RESULTS. Ultrasonography showed a sensitivity of 100%, a specificity of 52.6%, a positive predictive value of 100%, and a negative predictive value of 100% in regard to the indication for surgery in cases of surgical abdominal emergency. CONCLUSION: Ultrasonography saves time and money, can be performed in the emergency department, shows high sensitivity and specificity, and is the method of first choice in the evaluation of blunt trauma. PMID- 8712620 TI - [Role of microvascular free flaps in the treatment of amputation stumps]. AB - The authors report the importance of plastic surgery in rehabilitation of the amputated stump. After having referred the tight dependence between amputated stump and prosthesis, they underline the importance of a trophic and painless stump characterized by a satisfactory "useful length". They report two cases in which these priorities have been satisfied by using latissimus dorsi free flap. The authors believe that better rehabilitative results can be achieved by applying the most recent techniques utilized in plastic surgery and particularly microvascular free flaps. PMID- 8712621 TI - [Effects of allopurinol on damage caused by ischemia and reperfusion of skeletal muscles: an in vivo spectroscopic analysis (31P-MR) in rats]. AB - The effect of allopurinol on energetic metabolism (reutilization of hypoxanthine) has been studied in vivo by mean of 31P-RM spectroscopy on skeletal muscle in the rat in conditions of ischemia and reperfusion. The treatment with allopurinol demonstrates of on benefit or phosphocreatine and ATP kinetics. These results outline that reutilization of hypoxanthine doesn't represent a protective mechanism of allopurinol on skeletal muscles. The role of hypoxanthine reutilization has to be investigated with further researches. PMID- 8712622 TI - [Role of lactose, arginine and lysine combination in fracture healing (an experimental study)]. AB - L-arginine and L-lysine are essential amino acids which seem to possess some properties able to influence bone fractures healing. In fact, they seem to be involved not only in the increase of intestinal calcium adsorption but also in collagen synthesis, in insulin and growth hormone secretion and in osteoblastic activation. So, an experimental in vivo model was carried out by using 50 adult rabbits which, under general anaesthesia, were submitted to an osteotomy of the left fibula. Animals were divided into 5 groups and were daily treated with a mixture of lactose, L-arginine and L-lysine or with the only lactose (control group) at the same dosage as recommended for humans. They were sacrificed after 15, 30, 40, 50 and 60 days for radiological and histological studies. The results of the study showed that the pharmacological mixture containing L-arginine and L lysine accelerates and ameliorates the healing processes and this positive effect was particularly evident from the 30th day after the osteotomy. We think that these results are linked not only to calcium metabolism but also to different biological properties which positively contribute to a good healing of bone fractures. PMID- 8712623 TI - [Complex extravascular dislocation of a guidewire during catheterization of the subclavian vein]. AB - A case is presented in which a guidewire, during subclavian catheterization in a 63 years old patient, became knotted outside the vein, behind the clavear head of the sternocleidomastoid muscle. The wire was easily removed by surgical approach and the post operative course was uneventful. The causes of this complication are discussed and a few main points are emphasized when the Seldinger technique is employed for subclavian vein catheterization. PMID- 8712624 TI - Solitary fibrous tumor of the perithyroid soft tissue. Report of a case simulating a thyroid nodule. AB - A case of solitary fibrous tumor (SFT) of the perithyroidal soft tissue is reported. Due to its proximity to the upper aspect of the right thyroid lobe, its slow growth and its ultrasound features, the tumor was confused with a thyroid nodule. Two fine needle aspirations (FNA) gave only scant non diagnostic material. For these and cosmetic reasons together with the fact that ultrasound exam detected two solid nodules in the left lobe, a thyroidectomy was decided. Examination of the surgical specimen showed a nodule that was well demarcated and, although entirely extrathyroidal, bulged from the thyroid profile in a niche of the upper medial aspect of the right lobe. On histologic, immunohistochemical, and ultrastructural examination the tumor fulfilled the criteria for a diagnosis of solitary fibrous tumor. Thus, SFT must be considered in the differential diagnosis of suspected thyroid nodules mainly when FNA gives only scanty non diagnostic material which is unusual in thyroid hyperplastic nodules. PMID- 8712625 TI - [Perforation of the sigmoid caused by ingestion of foreign body]. AB - Dating from October 1987 to January 1995 we have observed 24 patients who swallowed an extraneous object. Tree of them underwent a surgical operation whereas the remaining 21 were monitored endoscopically. Our team use to operate when the extraneous object, for its shape or dimension, comes to a stop along the intestine, causing a potential of existent complication. In 2 of the 3 cases above mentioned, the diagnosis came out from a abdomen x-ray which put into evidence a clothes peg stopped at level with the first jejunal ansa and an open pin which had performed an ileal ansa. As far as concerns the third case, we were compelled to operate without a preoperative diagnosis as the patient, mentally disabled, swallowed a sharpened, not x-rayed extraneous object (a toothpick). When swallowed, a toothpick usually causes lesions in the higher stretch of the digestive system. It is very difficult to spot the toothpick position for it slack x-opacity, even when the lesion caused by it is clearly visible. The sigma perforation, caused by the swallow of an object, rarely happens. Authors have reported a case to be pointed out for its pathogenetic originality. PMID- 8712626 TI - [Carcinoid of the pancreas. 2 case reports]. AB - Two cases of carcinoid tumors are reported: the most important characteristic of these neoplasms are that they are uncommon and are accidentally and difficulty diagnosed. Surgery, when possible, is the treatment of choice for the carcinoid tumors of the pancreas, but in unresectable tumors chemotherapy, radiotherapy and interferon are used. PMID- 8712627 TI - Effect of stenting on graft vascularization after laryngotracheoplasty. AB - The management of pediatric airway stenosis has been an important topic of debate over the last few decades as prolonged intubation has become more common in the neonate. Although many surgical options are available, most would agree that use of expansion techniques (laryngotracheoplasty) with cartilaginous grafting is the procedure of choice for the severely stenotic airway. Controversy persists, however, regarding the role of stents. Advocates feel that stents serve to counteract scar contracture and support the newly constructed airway. In contrast, recent studies suggest that stenting results in impaired wound healing and an increased complication rate. The present study addresses the effect of stenting on the vascularization of cartilaginous grafts in an animal model. Thirty-six New Zealand white rabbits were evaluated after laryngotracheoplasty with autogenous cartilage grafting. Half of the animals were stented with a 2-cm section of an endotracheal tube that was secured just below the glottis. Three rabbits from each group were then painlessly sacrificed on days 4, 6, 8, 10, 14, and 21. Gross and histologic comparisons of the stented and nonstented specimens revealed similar wound healing. Measurements of graft vascularization were obtained with a computerized image measurement program, and a comparison was made regarding the rate of vascularization. There was a statistically significant increase in the rate of vascularization in the stented group (mean 73% versus 55% at day 10; p < .05). This analysis suggests that stenting does not inhibit early wound healing and specifically graft vascularization after laryngotracheoplasty. PMID- 8712628 TI - Long-term follow-up of recurrent laryngeal nerve avulsion for the treatment of spastic dysphonia. AB - Long-term follow-up of 3 to 7 years is reported on 18 patients who had undergone recurrent laryngeal nerve avulsion (RLNA) for the treatment of adductor spastic dysphonia (SD). Data on neural regrowth after previous recurrent laryngeal nerve section (RLNS) are presented in 2 of these 18 patients. We introduced RLNA as a modification of standard RLNS to prevent neural regrowth to the hemiparalyzed larynx and subsequent recurrence of SD. We have treated a total of 22 patients with RLNA, and now report a 3- to 7-year follow-up on 18 of these 22 patients. Resolution of symptoms was determined by routine follow-up assessment, perceptual voice analysis, and patient self-assessment. Sixteen of 18, or 89%, had no recurrence of spasms at 3 years after RLNA as determined at routine follow-up. Two of the 16 later developed spasms after medialization laryngoplasty for treatment of weak voice persistent after the avulsion. This yielded a total of 14 of 18, or 78%, who were unanimously judged by four speech pathologists to have no recurrence of SD at the longer follow-up period of 3 to 7 years. Two of these 4 patients were judged by all four analysts to have frequent, short spasms. The other 2 were judged by two of four analysts to have seldom, short spasms. Three of 18 patients presented with recurrent SD after previous RLNS. At the time of subsequent RLNA, each patient had evidence of neural regrowth at the distal nerve stump as demonstrated by intraoperative electromyography and histologic evaluation of the distal nerve stump. One remained free of SD following RLNA, 1 was free of spasms at 4 years after revision avulsion but developed spasms after medialization laryngoplasty, and the final patient developed spasms 3.75 years after revision RLNA. Medialization laryngoplasty with Silastic silicone rubber was performed in 6 of 18, with correction of postoperative breathiness in all 6, but with recurrence of spasm in 3. Spasms resolved in 1 of these with downsizing of the implant. We conclude that RLNA represents a useful treatment in the management of SD in patients not tolerant of botulinum toxin injections. PMID- 8712629 TI - Autologous fat injection for unilateral vocal fold paralysis. AB - This study evaluates the efficacy of autologous fat injection for medialization of the paralyzed vocal fold. In 21 patients with unilateral vocal fold paralysis, autologous abdominal fat was injected into the thyroarytenoid muscle to achieve medialization. All patients were followed up with serial videolaryngoscopy and voice evaluation. At 2 months' follow-up, the voice was judged to be excellent in 10 patients, slightly breathy but significantly better than the preoperative voice in 6 patients, and markedly breathy in 4 patients. At 3 to 4 months' follow up, of the 10 patients with excellent results, 5 maintained an excellent voice, 3 had developed slight breathiness, and 1 had developed severe breathiness. Long term (6 to 12 months) results were available in 11 patients, and all of them maintained the same voice quality that was noted during the 3 to 4 months' examination. Magnetic resonance imaging of the larynx was obtained in 7 patients at intervals ranging from 1 to 7 months and compared to the baseline scan obtained at 1 week postoperative to assess the amount of fat remaining in the muscle. The images showed fat volume to persist, but a decrease in the fat signal was observed over time. The results suggest that the duration of medialization with autologous fat is variable, but appears to last at least 2 to 3 months. This loss of volume after 3 months seems to be due to absorption of the fat and possibly muscle atrophy. Autologous fat injection is relatively safe and easy to perform, and is an ideal method of temporary vocal fold medialization in patients in whom return of vocal fold function is expected. PMID- 8712630 TI - Treatment of chronic paranasal sinus pain with minimal sinus disease. AB - A common problem for otolaryngologists are patients who present with recurrent, persistent sinus pain that appears out of proportion to the findings on physical examination. Often these patients have a history of recurrent sinusitis that required antibiotics or surgical intervention. Many have had repeated surgical procedures because of this pain. Other common past medical histories may include allergic rhinitis, facial trauma, or dental disease. Patients who have experienced documented acute sinusitis in the past will often present de novo with similar symptoms, but lack any objective evidence of a new active sinus infection. However, the diagnosis of sinusitis is not clearly removed from the patient's or clinician's mind, and the patient is further frustrated by the lack of adequate diagnosis, treatment, and resolution of symptoms. These patients may or may not be experiencing an upper respiratory tract infection or allergy with nasal drainage. Often, they are emotionally distraught from recurrent and persistent pain, the lack of resolution of their symptoms, dependency on narcotics and other analgesics, multiple consultations with a variety of clinicians, and the impingement of their symptoms on employment, interpersonal relationships, and societal and family obligations. If sinusitis is not found to be present, the otolaryngologist must help the patient understand this point, reassure him or her that the otolaryngologist will still be vigilant for the development of sinusitis, and refocus the history and workup for some other cause of the recurrent and persistent paranasal pain. We review various treatment approaches to paranasal pains that are not the result of sinusitis. PMID- 8712631 TI - Residue of middle ear cholesteatoma after intact canal wall tympanoplasty: surgical findings at one year. AB - The rate of residual disease after surgery for acquired middle ear cholesteatoma was investigated in 167 ears of 164 patients who had undergone planned second look tympanoplasty by the intact canal wall technique. Overall, operative findings at the second stage revealed 65 cases of residual disease in 48 ears (29%). These consisted of 50 squamous pearls, 11 cases of the flat, open type, and 4 cases of the extensive type. The configuration of residual disease is closely related to the technical difficulty of eradication, since en bloc removal is much easier in the squamous pearl than in the open or extensive type, mainly because of the unclear margin with the surrounding tissues. The proportion of cases of the open type was greater in children than in adults, in pars tensa cholesteatoma than in pars flaccida cholesteatoma, and in severe primary middle ear disease than in moderate or mild disease, although these differences were not statistically significant. The extensive type occurred in 4 ears with severe primary disease, 3 of which were in children. These results support the value and importance of the staged procedure for middle ear cholesteatoma, particularly when operated on by the intact canal wall technique. PMID- 8712632 TI - Endonasal endoscopic closure of cerebrospinal fluid fistulas at the anterior cranial base. AB - This study reports our indications and limits for endonasal endoscopic closure of dural defects with a cerebrospinal fluid (CSF) leak at the anterior cranial base, and demonstrates our surgical technique. Fifty-three patients with CSF rhinorrhea were reassessed for the success rate of closure of the CSF leak. Surgery was successful in 98%, and 68% of fistulas were closed endoscopically. A free graft of autogenous mucoperiosteum of the inferior turbinate was the most frequently used tissue for defect closure. The endonasal endoscopic route proved relatively safe for the closure of dural tears, irrespective of the cause, up to about 10 x 10 mm. It is characterized by minimal morbidity because of the preservation of sinus ventilation and bony structures, supraorbital nerves, and olfactory fibers. Defects larger in size, predominantly of traumatic origin, were closed via the transfacial approach. The decision on the surgical approach was additionally based on the extent of the facial soft tissue injuries and the localization of the leak. PMID- 8712633 TI - Microbiology of tracheal granulation tissue associated with silicone airway prostheses. AB - Granulation tissue formation and contraction is inevitable during the healing of open wounds. The presence of infection and a foreign body, such as a silicone airway prosthesis or other tracheotomy tube, magnifies this response. In airway reconstruction, wound contraction secondary to chronic inflammation is a liability. Cultures of granulation tissue were taken from 12 patients with silicone cannulas of T-Tubes. Polymicrobial colonization predominated, with variable combinations of gram-positive, gram-negative, and anaerobic organisms being present. On the basis of sensitivity data, amoxicillin-clavulanate potassium and ciprofloxacin hydrochloride were chosen as empirical antimicrobials. Anecdotally, impressive decreases in the quantity of granulation tissue were seen. We recommend a prospective trial to more accurately define the role of antibiotics in this setting. PMID- 8712634 TI - HLA class II-associated genetic susceptibility in idiopathic progressive sensorineural hearing loss. AB - To investigate the association between genes in the major histocompatibility complex and inner ear disease susceptibility at the DNA level, high-resolution genotyping for HLA class II (HLA-DR, -DQ, -DP) was performed by polymerase chain reaction-sequence-specific oligonucleotide reverse dot blot and polymerase chain reaction-restriction fragment length polymorphism analysis in 34 patients with idiopathic progressive sensorineural hearing loss (PSHL) and in 214 controls. The frequencies of DRB1*0301, DRB3*0101, DQB1*0201, and DPB1*0401 were significantly increased in patients with idiopathic PSHL compared with controls. The DQB1*0301 allele was significantly decreased in the patients. A linkage disequilibrium was probably responsible for the concomitant increase of both DRB1*0301 and DRB3*0101 alleles in patients. The increase of DQB1*0201 in patients was associated with the DRB1*0301 allele. In addition, the telomeric DPB1*0401 allele may act as an independent risk factor. The DQB1*0301 allele may have a protective role in the pathogenesis of idiopathic PSHL. These results suggest that the specific HLA class II gene products may confer susceptibility or resistance to idiopathic PSHL. PMID- 8712635 TI - Circulating intercellular adhesion molecule-1 and its cellular expression in head and neck non-Hodgkin's lymphomas, including lethal midline granuloma. AB - Extranodal non-Hodgkin's lymphoma in the head and neck, especially T-cell lymphoma of the lethal midline granuloma (LMG) type, has unique clinical and histologic features differentiating it from other lymphomas. The authors measured soluble intercellular adhesion molecule-1 (ICAM-1) in sera from 12 patients with T-cell lymphoma of the LMG type and from 52 patients with other head and neck non Hodgkin's lymphomas, by double-determinant immunoassay. The expression of ICAM-1 in lymphoma tissue was examined in 26 patients by the avidin-biotin immunoperoxidase method. The serum ICAM-1 levels were significantly higher in T cell lymphoma of the LMG type than in other head and neck lymphomas or in healthy adult controls. Elevated levels of serum ICAM-1 were associated with increased levels of serum lactate dehydrogenase, poor prognosis, and impaired T-cell dependent immune functions, both in T-cell lymphoma of the LMG type and in other head and neck lymphomas. When we monitored serum ICAM-1 levels in individual patients, the level decreased in the complete remission interval compared to before treatment and went up again when the lymphoma relapsed. Although the staining intensities of ICAM-1 in lymphoma cells were not related to serum ICAM-1 levels, a markedly intense expression of ICAM-1 was found on the angiocentric or angiodestructive lymphoreticular infiltrate region in the tissues from T-cell lymphoma of the LMG type. A higher serum ICAM-1 level and its tissue expression in T-cell lymphoma of the LMG type may be one of the clues to understanding this particular lymphoma. The serum ICAM-1 level could be an efficient parameter for monitoring the clinical course of head and neck non-Hodgkin's lymphomas. PMID- 8712636 TI - Secretory otitis media in adults: I. The role of mastoid pneumatizationas a risk factor. AB - We analyzed clinically 102 ears with secretory otitis media (SOM) belonging to 72 adult patients who during their adult life had not suffered previously from ear disease. As in children, most of the cases (63%) could be traced directly to an upper respiratory tract infection. The most striking finding was the preponderance of poorly pneumatized mastoids-which were measured planimetrically among our SOM cohort. This was found in adult SOM ears compared to contralateral healthy ears (4.59 versus 7.88 cm2), as well as when all 102 SOM ears were compared with values of the normal population (5.41 versus 12.9 cm2). This study showed that poorly pneumatized mastoids are a significant risk factor as far as adult SOM is concerned. PMID- 8712637 TI - Role of substance P in the vascular response of nasal mucosa in nasal allergy. AB - The effects of topically administered substance P (SP) on nasal blood flow and nasal airway resistance (NAR) were evaluated in 11 subjects with perennial nasal allergy. The change in NAR induced by SP was compared with those induced by nasal challenge with histamine, leukotriene D4 (LTD4), and antigen. In doses > or = 16 nmol, SP caused a significant increase of nasal blood flow within 5 minutes that lasted for less than 20 minutes. In doses > or = 16 nmol, SP caused a dose dependent, short-lasting, significant increase in NAR. The magnitude of the increase in NAR was LTD4 > SP > histamine when compared on a molar basis. Our results may suggest that SP released from C fiber terminals is partially involved in an early nasal vascular response after antigen challenge by acting on adjacent vascular smooth muscle to cause a transient vasodilatation of both resistance and capacitance vessels only while sensory stimulation persists in subjects with nasal allergy. PMID- 8712638 TI - Otoneurological expert system. AB - An otoneurological expert system was developed to help collect data and diagnose both central and peripheral diseases causing vertigo. Patient history and otoneurological and other examination results are used in the reasoning process. The case history data can be either mandatory or supportive. Mandatory questions are used to confirm a diagnosis, and conflicting answers are used to reject an unlikely disease. Supportive questions support or suppress a diagnosis, but their presence is not obligatory. The reasoning procedure of the otoneurological expert system scores every question independently for different diagnoses, depending on how well they agree with the symptom entity of a disease. Diagnostic criteria are set for each disease. Graphic displays illustrate the linear and nonlinear correlation between the symptoms and diseases. Emphasis is placed on diminishing the possibility of a wrong decision rather than maximizing the likelihood of reaching only one right decision, so that even rare diseases can be taken into consideration. PMID- 8712639 TI - High-resolution computed tomographic evaluation of the cochlear capsule in otosclerosis: relationship between densitometry and sensorineural hearing loss. AB - Otosclerotic cochlear involvement is a rather frequent disease that has not been clearly understood in terms of diagnosis and management. Objective evaluation methods are needed to confirm the clinical diagnosis, investigate the relationship with hearing impairment, and validate the results of treatment. In this study two ear groups with bone conduction hearing loss (BCHL) were investigated with audiometry and high-resolution computed tomography (HRCT). In the first group (n = 22) the diagnosis of fenestral otosclerosis was confirmed at operation; the second group (n = 9) was composed of ears clinically suspicious for purely cochlear involvement. Additionally, a control group (n = 14) of otologically normal ears was also studied. Foci of demineralization were demonstrated in 58% of the ears in the two groups; the sensorineural hearing loss (SNHL) in those ears was significantly worse than in those with normal radiologic findings. Three methods of HRCT densitometry were used to determine the abnormal regions in the cochlear capsule; the results suggested that hypodense regions were consistent with a greater degree of SNHL, in contrast to the hyperdense ones in ears with better cochlear reserves. Agreement was found between the location of the density change and the frequency topography of the SNHL; densitometric values were correlated with the bone conduction thresholds for certain frequencies. It is concluded that the spongiotic foci are responsible for the SNHL, since there was a correlation between their location and the SNHL frequency. The determination of better hearing in those ears with sclerotic foci supports the hypothesis that the sclerotic phase may not be a healing process following the spongiotic phase, and that it can be the first stage of the disease. PMID- 8712640 TI - Metastatic breast carcinoma presenting as cavernous sinus syndrome. PMID- 8712641 TI - Hypopharyngeal dilatation in a musician. PMID- 8712642 TI - Pharyngeal hypophysis. AB - Derived from embryologic remnants of Rathke's pouch and its craniopharyngeal duct, the pharyngeal hypophysis has long been considered to be present in all humans. This thesis has been recently challenged, but the weight of investigative evidence still favors the original premise. The pharyngeal hypophysis is just one product of Rathke's pouch. Others are extracranial pituitary tumors, the very rare craniopharyngioma, pharyngeal hypopharyngeal cysts, and cerebrospinal fluid rhinorrhea. PMID- 8712643 TI - Management of haemothorax. PMID- 8712644 TI - 'Emergency room' thoracotomy: is it ever justified? PMID- 8712645 TI - Thrombophilia and the surgeon. PMID- 8712646 TI - Twelve years experience of vena cava filtration. AB - Fourteen patients have required vena caval interruption in a period of 12 years at St Thomas' Hospital. Half of these patients had an underlying malignancy. None of these patients have had clinical or lung scan evidence of recurrent emboli. There were no deaths related to filter insertion and no patient died of a pulmonary embolism. Vena caval interruption is a procedure that is rarely necessary but may be valuable in carefully selected patients. PMID- 8712647 TI - Audit of 149 consecutive carotid endarterectomies performed by a single surgeon in a district general hospital over a 12-year period. AB - Carotid endarterectomy has been established by two large randomised controlled trials (European Carotid Surgery Trial (ECST) and North American Symptomatic Carotid Endarterectomy Trial (NASCET)) as an important surgical procedure for the prevention of ischaemic strokes in patients presenting with transient cerebral ischaemia or non-disabling strokes attributable to severe ipsilateral carotid artery stenosis. The operation carries significant risk of death and stroke and it has been advocated by some that carotid endarterectomy should only be performed in a small number of designated regional centres in order to achieve good surgical results. It is doubtful that the regional centres alone can cope with the increasing numbers of patients requiring carotid endarterectomy and there is therefore a requirement for the procedure to be carried out by vascular surgeons in district general hospitals. It is important that surgical results are audited to ensure that comparable outcomes are achieved. We present an audit of our experience of carotid endarterectomy since 1981. A total of 149 consecutive carotid endarterectomies were performed by a single surgeon with a special interest in carotid surgery. The results are comparable to ECST with a 30-day mortality of 0% and an overall 30-day stroke rate of 5.7% (major strokes) for patients with severe, ie 70-99%, ipsilateral carotid artery stenoses. We have shown that carotid endarterectomy is an operation that can be performed safely and with good results by suitably trained surgeons in district general hospitals. PMID- 8712648 TI - Morbidity of varicose vein surgery: auditing the benefit of changing clinical practice. AB - During two consecutive study periods (24 months and 16 months), 997 patients (1322 legs) with varicose veins underwent surgical treatment in Huddersfield Royal Infirmary, using a variety of surgical techniques. The average length of stay was 1.5 days, and 95.5% of patients had a hospital stay of less than 2 days. Complications occurred in seven inpatients. A further 16 patients developed complications requiring readmission to hospital (10 minor and intermediate; 6 major). The complication rate appeared to be operator-dependent: an increased complication rate (particularly major complications) occurred after surgery by junior surgeons. Major complications included femoral vein injury (1 patient), postoperative deep venous thrombosis (4 patients), pulmonary embolism (1 patient) and groin lymphatic fistula requiring reoperation (1 patient). Between the first and second study period, a change in policy regarding the type of bandaging used and the use of postoperative antiembolic stockings occurred and appears to have reduced the incidence of thromboembolic complications from 0.7% to 0.2%. Varicose vein surgery is not without major complications though, fortunately, there was no mortality in this series. These complications can be minimised with good surgical technique and better supervision of surgical trainees. Modification of postoperative management can further enhance the safety of this procedure. PMID- 8712649 TI - Pathogenesis of pancreatic infection. AB - John Hunter studied comparative anatomy of the pancreas but was unaware of pancreatic infection which is now the leading cause of mortality in pancreatitis. This was investigated using a feline model of pancreatitis. Pathogens spread to the healthy and inflamed gland from many sources including colon, gallbladder, or a septic focus and by various routes including the circulation, reflux into the pancreatic duct or by transmural migration from the colon. Colonisation risk was proportional to necrosis and inflammation, confirming clinical observations. These studies showed that pathogens frequently colonised the pancreas, but infection developed only in animals with pancreatitis. In cats with pancreatitis, phagocytic function was reduced by 28%. This was probably owing to phagocytic capacity being overwhelmed by protease-antiprotease complexes because, in humans, granulocyte and lymphocyte function was normal. These experiments suggested that it would be difficult to prevent pancreatic colonisation, but indicated some types of therapy may have potential. These were investigated using this animal model of pancreatic infection. Treatment with either cefotaxime or levamisole (an immunostimulant) were effective. However, the anti-inflammatory drug dopamine, which reduced inflammation, did not eradicate all pathogens. PMID- 8712650 TI - Parathyroid hormone-related protein, bone metastases and hypercalcaemia of malignancy. AB - Breast cancer patients frequently develop bone metastases. A newly discovered peptide, Parathyroid Hormone Related Protein (PTHrP), causes humoral hypercalcaemia of malignancy. We have studied whether production of this protein by breast cancers leads to the development of bone metastases or hypercalcaemia. PTHrP was produced by nearly 60% of early breast cancers and its production by the tumours was associated with malignant mammographic microcalcification, and the development of both bone metastases and hypercalcaemia. The hypercalcaemia associated with bone metastases has been shown to have a significant humoral component and measurement of plasma PTHrP in patients with hypercalcaemia is diagnostically useful. Potential mechanisms of preventing bone metastasis are discussed. PMID- 8712651 TI - Laparoscopic hernia repair in Leicester General Hospital: a prospective audit of 94 patients. AB - Conventional hernia repair is effective in terms of cure but is associated with considerable postoperative pain and delay in return to normal activity. Laparoscopic repair has the potential to reduce pain and speed return to normal activity, but there have been few published reports of the outcome of this operation in the UK. We present a prospective audit of 94 patients who underwent laparoscopic repair. Of the 94 patients, 87 (92.6%) were male and 7 (7.4%) were female. Thirteen of the repairs were bilateral and 12 were recurrent. Two had to be converted to open repair. The mean operating time for unilateral repair was 56 min and for bilateral repair 98 min. Sixty-three patients (67%) were discharged within 24 h and 21 (22.4%) were discharged within 48 h. There were minor complications in 20 patients (21%), eight of whom (8.5%) developed a haematoma. The other minor complications included seromas (2), bruising at the site of the entry port (2), hyperaesthesia in the groin (2), port hernia (1), shoulder tip pain after surgery (3) and postoperative urinary retention (2). Nine (9.5%) patients claimed to have had no pain or discomfort at all; 35 (37.2%) were pain and discomfort free in 2 weeks. Thirty-two (34%) patients returned to normal activities in 2 weeks. With a median follow-up of 8 months 3 (3.2%) recurrences were noted. It is emphasised that this series represents a learning curve and that the operation is developmental. We are now restricting laparoscopic repair to recurrent and bilateral hernias where the technique offers particular advantages. PMID- 8712652 TI - Comparison of diclofenac sodium and morphine sulphate for postoperative analgesia after day case inguinal hernia surgery. AB - Postoperative pain may be a significant reason for delayed discharge from hospital, increased morbidity and reduced patient satisfaction with ambulatory hernia surgery. This study compared two postoperative oral analgesic protocols after day case inguinal hernia repair; 30 mg morphine sulphate (MST) and 10 mg metoclopramide every 8 h for 48 h or 75 mg diclofenac twice daily for 48 h. The pain reported in the MST group was significantly greater on both the day of operation and the first postoperative day (P < 0.05, Mann-Whitney U test). A significantly higher proportion of patients taking MST complained of nausea on the day of operation and on the 1st postoperative day (P < 0.05, chi 2). The time taken to walk, dress and leave home alone were achieved in a significantly shorter duration in patients taking diclofenac. We conclude that diclofenac provides effective analgesia, has a more acceptable side-effect profile than morphine sulphate and is the treatment of choice after ambulatory hernia surgery. PMID- 8712653 TI - Inguinal hernia and a single strenuous event. AB - A study of 129 consecutive patients, who had a total of 145 inguinal hernias, showed that in only 7% of the patients was the hernia subjectively attributable to a single muscular strain. Guidelines are suggested to assist in assessing 'cause' in claims for industrial injury in such patients. PMID- 8712654 TI - Impact of a restorative dentistry service on the prescription of apical surgery in a district general hospital. AB - The case records of a group of patients who had undergone periapical surgery in a district general hospital were examined and compared with the criteria agreed by a group of hospital restorative dentists as to whether apical surgery was appropriate rather than, at least initially, a non-surgical treatment option. Only 65% of the patients met the audit criteria. As a result, changes have been made to clinical practice, including a greater involvement of restorative clinicians in the decision-making process when apical surgery is under consideration. PMID- 8712656 TI - Audit of pain after nasal surgery. AB - In our department, nasal packs have traditionally remained in place for a period of 24 h after routine elective nasal surgery. We decided to determine whether reducing the duration of nasal packing from 24 h to 2 h would reduce postoperative pain without a concomitant increase in haemorrhagic complications. A series of 72 patients undergoing elective nasal surgery were randomised to have either nasal packs for 24 h (n = 39) or for 2 h (n = 33). Pain was assessed using a visual analogue scale with the nasal packs in place and on their removal. The incidence of haemorrhage was noted. The results showed a significant decrease in mean pain score (MPS) with the packs in place, from 3.4 (range 0.8-6.8) to 1.16 (range 0-2.5); (P < 0.001) and also on pack removal from 5.3 (range 1.2-7.9) to 2.6 (range 0-5.4); (P < 0.05). There was no postoperative haemorrhage in either group. A new standard of 2 h for routine nasal packing was set and our departmental protocols were changed accordingly. Results in a further 36 patients gave a MPS with the packs in place of 1.12 (range 0-3.2) and a MPS of 2.36 (range 0-6.7) on their removal. There was no statistical difference between these and our previous findings. We conclude that postoperative nasal packing for only 2 h rather than 24 h significantly reduces pain without a concomitant risk of haemorrhage. PMID- 8712655 TI - Nature and mechanism of peripheral nerve damage in an experimental model of non freezing cold injury. AB - Non-freezing cold injury (NFCI), so called trench foot, is a condition characterised by a peripheral neuropathy, developing when the extremities are exposed for prolonged periods to wet conditions at temperatures just above freezing. Classically, military personnel are affected, with 14% of casualties in the Falklands conflict afflicted. Clinically, NFCI is characterised by a well defined acute clinical picture and chronic sequelae. Little is known regarding the pathophysiology and treatment of this condition. Opinions vary as to the type of nerve fibres most susceptible to damage and proposed mechanisms of injury include direct axonal damage, ischaemia and ischaemia/reperfusion. A series of investigations has been performed to clarify which populations of nerve fibres are more susceptible to damage, and to elucidate the exact mechanism of nerve injury. An in vivo rabbit hind limb model, subjected to 16 h of cold immersion (1 2 degrees C), provided the basis of this study. Nerve specimens were examined by semi-thin sectioning for myelin fibre counts, by electron microscopy to assess the unmyelinated fibre population, and fine nerve terminals in plantar skin were assessed immunohistochemically. The results showed that large myelinated fibres were preferentially damaged, while small myelinated and unmyelinated fibres were relatively spared. Nerve damage was found to start proximally and extend distally with time. Serial temperature measurements identified a warm-cold interface in the upper tibial region of immersed limbs. As this was the initial site of injury, this suggested that a dynamic balance exists in the cold immersed limb between the protective effects of cooling and the damaging effects of ischaemia. The non-invasive technique of near infrared spectroscopy was used to measure changes in tissue oxygen supply and utilisation and blood volume. The findings supported the hypothesis that an interface is created at the site of initial nerve damage in the upper tibia, where cyclical ischaemia-reperfusion injury occurs. PMID- 8712657 TI - An effective shield for free: pulsed lavage in total knee replacement. PMID- 8712658 TI - 'Above and below' dilatation of anastomotic colorectal strictures. AB - A new technique is described for dilating anastomotic colorectal strictures lying distal to a protecting stoma where attempts at instrumentation from below have failed. The new manoeuvre consists of approaching the stricture simultaneously from above, using a fibrescope inserted through the stoma, and from below using a rigid sigmoidoscope. A guidewire is passed from above through the stricture and retrieved by the lower operator. Dilators can then be railroaded through it. Six successful cases are reported. PMID- 8712659 TI - A simple method of achieving rapid haemostasis in PTFE grafts after on-table angiography. PMID- 8712660 TI - Laparoscopic repair of perforated duodenal ulcers with a falciform ligament patch. AB - Six patients are reported who presented with perforated duodenal ulcers. These were repaired laparoscopically by suturing the falciform ligament over the perforated ulcer. There were no complications and patients were discharged between 3 and 5 days after surgery. This technique is simpler to perform than a laparoscopic omental patch repair. PMID- 8712661 TI - Injection into the synovial space of the flexor tendons of the hand. PMID- 8712662 TI - Removal of an inhaled needle from the mediastinum with the aid of a metal detector. PMID- 8712663 TI - Antipersonnel mines: the global endemic. PMID- 8712664 TI - Landmine injuries. PMID- 8712665 TI - Subjective effects of double gloves on surgical performance. PMID- 8712666 TI - Four-year evaluation of a direct-access fibreoptic sigmoidoscopy service. PMID- 8712667 TI - Use of skin staples for securing the mesh in the Lichtenstein repair of inguinal hernia. PMID- 8712668 TI - Use of skin staples for securing the mesh in the Lichtenstein repair of inguinal hernia. PMID- 8712669 TI - Use of skin staples for securing the mesh in the Lichtenstein repair of inguinal hernia. PMID- 8712670 TI - Iatrogenic accessory nerve injury. PMID- 8712671 TI - Minimum standards for the management of cleft lip and palate: efforts to close the audit loop. PMID- 8712672 TI - Problematic renal calculi presenting during pregnancy. PMID- 8712673 TI - Iatrogenic accessory nerve injury. PMID- 8712674 TI - Endoscopic stapling diverticulotomy of pharyngeal pouch. PMID- 8712675 TI - An audit of one surgeon's experience of oral squamous cell carcinoma using a computerised malignancy database. PMID- 8712676 TI - Cancer: an historical perspective. PMID- 8712677 TI - Relationship between topoisomerase II levels and resistance to topoisomerase II inhibitors in lung cancer cell lines. AB - Topoisomerase II is a key target of many anticancer drugs used to treat lung cancer. We measured the expression of topoisomerase II alpha and beta mRNA's and also the levels of cellular topoisomerase II alpha and beta protein and concluded that topoisomerase II alpha levels are important in cellular resistance to the topoisomerase II inhibitors examined. This can be clearly seen in pairs of matched cell lines. However, when looking at a panel of cell lines with a range of histological types the importance of the enzyme can be masked by other cellular characteristics such as repair and detoxification mechanisms. PMID- 8712678 TI - Adenovirus-mediated thymidine kinase gene transduction in human epithelial ovarian cancer cell lines followed by exposure to ganciclovir. AB - In an effort to develop gene therapy for ovarian cancer efficacy and toxicity of adenovirus-mediated transfer of the HSV-TK gene followed by administration of ganciclovir were studied in two human epithelial ovarian cancer cell lines Ov-ca 2774 and Ov-ca-1225. 100% transduction was achieved in both cell lines at MOIs of 7 and 15 as demonstrated by X-Gal staining. No toxicity of virus alone was observed at MOIs up to 30. GCV was not toxic up to 200 micrograms/ml. Cell killing efficacy was shown to be dependent on MOI as well as GCV dose. The "bystander effect" of ADV/RSV-TK was quantified by mixing experiments and found to be dependent on the proportion of ADV/RSV-TK positive cells as well as the GCV dosage. Similar results were observed in both cell lines. ADV/RSV-TK mediated gene therapy may be a promising approach in ovarian cancer. PMID- 8712679 TI - Antitumour activity of photosensitized hypericin on A431 cell xenografts. AB - The in vivo antitumour activity of the natural photosensitizer hypericin was evaluated. Athymic nude mice xenografted with A431 cells were intraperitoneally administered with different hypericin doses and the tumours were locally irradiated 2 h later with white light (180 J/cm2) using a cold light source. When treatment was started one day after tumour inoculation, a dose-dependent antitumour effect was observed in light-treated animals. Complete inhibition of the tumour growth was achieved with 2.5 mg/kg hypericin. When the efficacy of a single hypericin dose (5 mg/kg) followed by a single light treatment on established tumours (60 mm3) was investigated, an 80% reduction in tumour mass was seen. Furthermore, an accumulation of the photosensitizer in A431 xenografts was observed after local light irradiation. Our results strongly suggest that hypericin holds promise as a new, safe, efficient and selective PDT photosensitizer. PMID- 8712680 TI - Cysteine proteinase inhibitors reduce malignant melanoma cell invasion in vitro. AB - Proteolytic enzymes are believed to be necessary for tumor cell invasion. We have studied the effects of the cysteine proteinase inhibitor E-64 and the serine and cysteine proteinase inhibitor leupeptin, on the ability of human malignant melanoma cells (LOX) to pass through an artificial basement membrane. Transwell chambers containing filters coated with the reconstituted basement membrane, Matrigel, were used. Nontoxic concentrations of the proteinase inhibitors reduced the invasion of LOX cells through Matrigel. E-64 (250 mumol/l) by 27% and leupepetin (250 micrograms/ml) by 46%. The proteinase inhibitors did not alter the growth rate of the tumor cells, their motility through uncoated filters, or their attachment to the Matrigel coated wells. Our results indicate that cysteine proteinases are involved in the degradation of basement membranes and thus contribute to the invasion of malignant melanoma cells. PMID- 8712681 TI - Effects of TNF-alpha and retinoic acid on the proliferation of the clear cell and chromophilic types of human renal cell carcinoma in vitro. AB - Since no effective therapeutic approach is known so far for metastatic renal cell carcinoma (RCC), we analyzed the anti-proliferative effects of TNF-alpha and retinoic acid (RA), applied either alone or in combination on 7 different RCC cell lines in vitro. In 5 out of 7 cell lines, a significant (p < 0.05) dose dependent inhibition of tumor cell proliferation became evident after exposure to TNF-alpha, the response being of modest magnitude in 5 cell lines. In 2 cell lines the effects were more pronounced with a reduction of cell viability to 55 +/- 11% of the control. Northern blot analysis revealed no expression of TNF alpha and p75 TNF-receptor in any cell line. All the cell lines showed p55 TNF receptor mRNA. Scatchard analysis revealed no correlation between TNF-alpha receptor status and growth inhibitory response to TNF-alpha, the number of TNF receptors per cell ranging from 0 to 3,976, and the affinity values from Kd = 0.621 nmol/l to Kd = 4.28 nmol/l. Exposure to RA alone resulted in significant (p < 0.05), but modest growth inhibition in 2 out of 7 cell lines with a reduction of cell viability to 83 +/- 1% of the control. In 2 out of 7 cell lines, combination of RA and TNF-alpha was significantly (p < 0.05) more effective than the single application of each compound. Northern blot analysis revealed no transcripts of CRABP I and retinoic acid receptor (RAR)-beta. All the cell lines expressed RAR-alpha mRNA and one cell line additionally expressed RAR-gamma mRNA. A correlation between RAR status and RA response was not seen. PMID- 8712682 TI - Modulation of calcium signalling and proliferation in monoblastoid U-937 cells. AB - The effects of TPA (12-0-tetradecanoylphorbol-13-acetate) and G-protein modulators on the concentration of cytoplasmic Ca2+ ([Ca2+]i), cytoplasmic pH and cell growth were investigated in monoblastoid U-937 cells. The G-protein activator NaF causes a dose-dependent increase of [Ca2+]i, that is partially sensitive to inhibition by pertussis toxin. The [Ca2+]i rise appears to come mainly from extracellular sources, and the Ca2+ influx is mediated by channels insensitive to the Ca2+ blocker verapamil. The Ca2+ ionophore ionomycin causes a biphasic rise of [Ca2+]i, reaching steady state levels slightly higher than those attained with NaF. TPA per se has no effect on [Ca2+]i, but potently reverses the NaF or ionomycin induced [Ca2+]i rise. Also, TPA partially counteracted the acidification induced by NaF. Both NaF and ionomycin per se had no effect on cell growth but partially counteracted TPA induced growth inhibition. Interferon-gamma and tumor necrosis factor-alpha did not affect [Ca2+]i by themselves but lowered the [Ca2+]i of NaF stimulated cells. The cytokines had no effect on cytoplasmic pH. This study indicates that elevations of [Ca2+]i in themselves does not trigger proliferation, but alterations of [Ca2+]i modulates the regulation of U937-cell growth. PMID- 8712683 TI - Isolation and identification of selenium-labeled proteins in the mouse kidney. AB - Selenium is a potent chemopreventive agent; however, the mechanisms for its chemopreventive activities remain elusive. Selenium binds to several proteins, some of which require selenium for functional activity. In this study, two 58kDa selenium-labeled proteins were identified in mouse kidney using a 75Se labeling method. The proteins were partially purified using Sephadex G.150 gel filtration, DEAE-Sephadex A-50 ion-exchange chromatography and one- / two-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis (1D-/2D-SDS-PAGE). The two proteins migrated at 58kDa on 2D-SDS-PAGE and differed only slightly in their pI values; i.e., 6.2 and 6.6, respectively. The polyclonal antibodies raised in rabbits against the 58kDa proteins electro-eluted from the 1D-SDS-PAGE of the DEAE purified fraction, recognized both protein spots on 2D-SDS-PAGE gel. The in situ enzymatic digestion of the two proteins separated in 2D-SDS-PAGE gels, followed by microsequencing of the peptides, resulted in the identification of these two proteins as related to human lipoamide dehydrogenase and thiol: protein disulfide oxidoreductase (TPDO). In common, both these proteins have a bis (cysteinyl) sequence motif cys-X-X-cys (for lipoamide dehydrogenase it is cys-X-X X-X-cys) which is also an integral part of several other proteins such as thioredoxin, protein disulfide isomerase, endoplasmic reticulum protein (ERp72), selenoprotein W, 56kDa acetaminophen binding protein and formate dehydrogenase. This sequence motif acts as an active redox center for majority of the proteins mentioned above, that may be controlling the oxidation/reduction of proteins in vivo. How and why selenium is binding to proteins with this common sequence motif needs further investigation. PMID- 8712684 TI - Tumorigenicity, invasion and metastasis of the small cell lung cancer cell line NCI-H69 and two derivative lines MOG-H69V and MOG-H69VZ. AB - Two adherent cell lines MOG-H69V and MOG-H69VZ have been isolated from a continuous cell line, NCI-H69, derived from human small cell lung cancer by Carney et al, [1987]. They have been established and characterised morphologically, biochemically, and for growth characteristics in vitro Khan et al (19). In the present study both the parental and the derivative lines have been investigated for invasiveness in vitro and in vivo. The parental line showed an early invasiveness compared with both the derivative cell lines. All cell lines formed tumours in nude mice with 100% take rate. Xenograft histology of all the cell lines revealed pleomorphic tumours, however the derivative lines showed areas of focal, large, spindle cells containing both acidic and neutral mucin, and spaces between the cells were found filled with alcianophilic, amorphous material. The parental line was invasive and metastatic. Tumours of both the derivative lines were non-metastatic under similar conditions. They were also investigated for neuroendocrine-cell marker expression. These data show that while the behaviour of the parental line was compatible with small cell lung cancer, that of the derivative lines was more indicative of non-small cell lung cancer, both in vitro and in vivo. As previous data suggested a common origin of the parental and the derivative lines, probably from a stem cell subpopulation present in the parental line, these lines represent a useful model for the study of phenotypic changes in lung cancer. PMID- 8712685 TI - p53 accumulation in various organs of rats after whole-body exposure to low-dose X-ray irradiation. AB - We report the induction of wild-type p53 protein accumulation in various organs within 12 h of exposure to X-ray irradiation at low doses (10, 25 or 50 cGy) in rats. The levels of p53 in some organs of irradiated rats were increased about 2- to 3-fold in comparison with the basal p53 levels in non-irradiated rats. Differences in the levels of p53 induction after low-dose X-ray irradiation were observed in the small intestine, bone marrow, brain, liver, adrenal gland, spleen, hypophysis and skin. In contrast, there was no obvious accumulation of p53 protein in the testis and ovary. Thus, the induction of cellular p53 accumulation by low-dose X-ray irradiation in rats seems to be organ-specific. We consider that cell type, and interactions with other signal transduction pathways of the hormone system, immune system and nervous system may contribute to the variable induction of p53 by low-dose X-ray irradiation. PMID- 8712686 TI - CG5/Dx human breast cancer cell line: characterization of a new doxorubicin resistant variant. AB - By continuous exposure of CG5 human breast cancer cell line to increasing doxorubicin (Dx) concentrations, a multidrug-resistant (MDR) subline (CG5/Dx) was obtained. The resistant variant showed P-glycoprotein (P-gp) expression and a lower intracellular doxorubicin level than the parental cells. CG5/Dx cells were 19.4 fold more resistant to Dx than CG5 cells and showed a cross-resistance to some structurally related and unrelated compounds. Differences in kinetics, biological and ultrastructural features between the two cell lines were investigated. The CG5/Dx cells grew more slowly, produced higher CEA levels and showed a reduced progesterone receptor (PgR) content than the parental cells. Ultrastructural studies revealed differences involving, polyribosomes, rough endoplasmic reticulum, [mitochondria] and cytoskeleton. PMID- 8712687 TI - Functional inactivation of p53 by antisense RNA induces invasive ability of lung carcinoma cells and downregulates cytokeratin synthesis. AB - Previously we had identified a p53 DNA-binding motif in the 5' region of the CK8 gene This finding led us to study the role of p53 protein in the regulation of CK8 gene expression and its role in tumorigenesis. Human lung cancer cell lines stably transfected with antisense p53 cDNA that expressed little p53 protein were analyzed. CK8 mRNA and the protein expression in these p53 antisense clones were very low as revealed by northern and western blot analysis. Immunofluorescence studies indicated that the cytokeratin networks around the nuclei of these cells collapsed; although some staining was observed around the nuclei. Antisense clones were highly invasive on in vitro matrigel assay. Scanning electron microscopy of the surface topography of these antisense clones revealed a large number of microvilli on their surfaces, a phenotype characteristic of tumor cell invasion. These findings suggest that functional inactivation p53 protein could be an important step in tumor progression. PMID- 8712688 TI - Mutant H-ras overexpression inhibits drug and U.V. induced apoptosis. AB - Various studies have shown that oncogene and oncosuppressor gene activity can enhance or suppress programmed cell death (apoptosis) in various cell systems. Recent data indicates that overexpression of activated H-ras could influence that onset of apoptosis. We investigated the role of activated H-ras in the apoptotic cell death of rat fibroblast lines. We found that forced overexpression of H-ras induced resistance to U.V. and drug induced apoptosis. We examined possible mechanisms for the action of H-ras in resistance to apoptosis. It was found that both ras transfected and ras untransfected lines displayed similar endonuclease activities. In addition, it was found that the irradiated ras transfected line showed inhibited production of peroxides compared to the irradiated ras untransfected line. Drug induced apoptosis did not appear to involve peroxide production. In addition the antioxidant compound PDTC, was found to inhibit U.V. induced apoptosis but not drug induced apoptosis. In addition, we found the ras transfected line to possess elevated levels of catalase compared to the parent untransfected line. Thus we suggest that an anti-oxidant mechanism, possibly mediated by forced overexpression of activated H-ras could protect cells from apoptosis. PMID- 8712689 TI - Molecular genetics of malignant insulinoma. AB - Malignant insulinoma is an rare form of cancer with poor prognosis and a reported 5-year survival of 35%. Relatively little is known about the etiology of this disease or of the oncogenes and tumor suppressor genes that participate in its genesis and progression. To address this issue, several protooncogenes, including K-ras, N-ras, erbB-2, erbB-3,c-myc, c-fos, c-jun were examined. Also analyzed was the expression of the growth factors TGF-alpha, EGF, and insulin as well as the EGF receptor (EGF-R), p53 and the putative anti-metastasis gene nm23-H1. These were examined in malignant insulinomas, benign insulinomas, pancreatic B cell hyperplasias and in normal endocrine pancreas. Normal endocrine pancreas showed moderate immunoreaction for c-myc and a strong reaction for insulin. All other parameters were negative. Benign pancreatic B cell hyperplasias were slightly or moderately positive for N-ras and TGF-alpha, and were weakly positive for EGF-R. They were strongly positive for c-myc and insulin. In malignant insulinomas there was strong immunoreaction for c-myc, TGF-alpha, N-ras, K-ras and p53. Insulin reaction was moderate or strong. Molecular genetic studies have been performed for the presence of activating point mutations in codon 12 of the c-K-ras oncogene. Mutations were detected using primer-mediated, mutant-enriched, polymerase chain reaction-restriction fragment length polymorphism analysis and were further characterized by allele-specific oligonucleotide hybridization. Four out of six patients with malignant insulinoma and two out of eight patients with benign insulinoma harbored K-ras point mutations at codon 12. All patients with mutated K-ras oncogene also had elevated levels of p53 protein as well as c-myc and TGF-alpha. In one extremely malignant case we found concomitant mutation at codon 12 of K-ras and codon 61 of the N-ras gene. Our data are consistent with the idea that malignant progression is accompanied by the progressive accumulation of multiple genetic lesions and suggest that activation of myc, TGF alpha and ras genes may be early events in the development of insulinoma. PMID- 8712690 TI - Influence of culture media and multidrug resistance on the wheat germ agglutinin (WGA) glycocytochemical expression of two human glioblastoma cell lines. AB - Over the last two decades, many studies carried out with the aid of lectins have firmly established that cell glycans usually change in the course of the normal processes of growth and development, as well as in pathological situations. We describe here the in vivo binding expression of wheat germ agglutinin (WGA) to the U87 and U373 human glioblastoma cell lines exposed to various culture media i.e., media supplemented with either 10% (FCS10) or 1% (FCS1) fetal calf serum with or without 10 n Mol/l 17 beta-oestradiol (E2). After exposure to chemotherapeutic agents, the resistant variants (CR) developed by the two cell lines were also investigated. The quantitative cytochemical assessment of WGA binding was assessed by means of a cell image processor, which was also used to determine ploidy level (on Feulgen-stained nuclei) by means of DNA histogram typing (DHT). Our results clearly demonstrate that when U373 cells are cultured with E2, this steroid can modify the expression of WGA binding, whereas U87 cells were unaffected. Similarly, lowering the FCS level enhanced the WGA binding of the U373 cell line. Multidrug-resistant cell variants were associated with both aneuploidy and a dramatic decrease in cytochemical WGA expression. PMID- 8712691 TI - Evidence for the release of autocrine growth factor(s) by Hodgkin's cell lines. AB - A cell surface glycoprotein (BLA.36) expressed on Hodgkin's cell and also on normal and malignant B lymphocytes, was identified by reaction with a monoclonal antibody. Expression of BLA.36 was detected on Hodgkin's cell lines, as well as B or pre-B cell lines, but was not detectable on other hematopoietic, carcinoma or melanoma cell lines, both by immunoprecipitation of intrinsically labeled cell lines and by immunocytochemical staining methods. Hodgkin's cell lines which were positive for BLA.36 expression were also responsive to a partially purified Hodgkin's cell growth factor(s) (HCGFs). HCGFs was partially purified by concentrating conditioned medium from a Hodgkin's cell line (HDLM-3), followed by ion-exchange chromatography (DE52) and gel filtration on Sephadex G-50. The active fractions yielded protein bands with apparent molecular weights ranging from 16 to 21 kilodaltons on sodium dodecyl sulphate-polyacrylamide gel electrophoresis under both non-reducing or reducing conditions. Hodgkin's cell lines treated with antiBLA.36 antibody were not growth responsive to exogenously added HCGFs. Addition of alpha-interferon (alpha-IFN) to BLA.36-positive Hodgkin's cell lines resulted in down-regulation of BLA.36, accompanied by inhibition of growth. Cell lines treated in this way failed to respond to exogenously added HCGFs, suggesting a requirement for BLA.36 expression in order for HCGFs to exercise its growth-stimulatory effect. PMID- 8712692 TI - Expression of alternatively spliced lck transcripts from the proximal promoter in colorectal cancer derived cell lines. AB - Ectopic expression of the lck gene has been demonstrated in epithelial tumor cells to be directed by the proximal promoter element. Variability of lck transcripts at the 5' end detected in T-cells and lymphoid malignancies prompted us to investigate the potential heterogeneity of lck mRNA molecules in colorectal cancer derived cell lines. Our experiments clearly indicate that the three different types of lck transcripts, which are exclusively derived from the proximal promoter, were expressed in our panel of colorectal cancer cells analyzed. DNA sequence analyses of the reversely transcribed lck mRNAs revealed that they indeed represented the lck transcripts designated type 1A, 1B and 1C, which have recently been identified in T-cells. Similarly with the situation in immature hematopoietic malignancies lck transcript 1B steady state levels were predominant in colorectal cancer derived cell lines. These findings lend further support to the notion, that expression of lck is part of anectopic lymphoid activation and differentiation program induced in these epithelial tumors. PMID- 8712693 TI - Inhibition of the development of rat liver preneoplastic lesions by verapamil and dexverapamil. AB - The effect of verapamil and dexverapamil on the development of liver preneoplastic foci was investigated in male Sprague-Dawley rats initiated with N nitrosodiethylamine (200 mg/kg i.p.), fed on a diet containing 0.01% 2 acetylaminofluorene and subjected to partial hepatectomy, according to the hepatocarcinogenesis model developed by Solt and Farber. Administration of drinking water containing 0.03% verapamil or dexverapamil resulted in a decrease in the incidence and size of gamma-glutamyl transpeptidase-positive foci. The chemopreventive effect of dexverapamil was more marked than that of verapamil. These findings support the hypothesis that these two calcium channel blockers act by reducing the resistance of initiated hepatocytes to the mitoinhibitory and cytotoxic effects of 2-acetylaminofluorene. PMID- 8712694 TI - Paclitaxel and cisplatin sensitivity of ovarian carcinoma cell lines tested with the 96-well plate clonogenic assay. AB - Platinum based chemotherapy is the cornerstone of treatment in advanced ovarian carcinoma. Paclitaxel, an unique antimicrotubule agent has shown significant clinical activity in cisplatin-resistant tumours, indicating a lack of cross resistance. To compare the in vitro sensitivity of ovarian carcinoma to cisplatin and paclitaxel, we tested 7 ovarian carcinoma cell lines with the 96-well plate clonogenic assay. Chemosensitivity was expressed as the IC50 value i.e. the drug concentration causing 50% inhibition of clonogenic survival. IC50 values were obtained from dose-response curves after fitting the data to the linear quadratic equation. The IC50 values for paclitacel were 0.4-3.4 nM, showing an 8.5-fold difference between various cell lines. The IC50 values for cisplatin were 0.1 0.45 ug ml-1 showing only a 4.5-fold difference. This variance is clearly smaller than the 25-fold difference observed with the same method in endometrial carcinoma cell lines (Rantanen et al, Br J Cancer 69: 482-86, 1994). In accordance with clinical findings, no cross-resistance or correlation between sensitivity to paclitaxel and cisplatin could be demonstrated. PMID- 8712695 TI - Anti free radical action of calcium antagonists and H1 and H2 receptors antagonists in neoplastic disease. AB - The blood of the subjects suffering from Neoplastic Disease (ND) shows phenomena of membrane peroxidation due to the presence of Free Radicals (FRs), in a quantity much greater than the one observed in the blood of healthy subjects. This can be detected either by calculating the time necessary for the formation of "Heinz bodies" (Hbs), (p < 0.00001) after oxidative stress of the blood in vitro with acetylphenylidrazine (APH), or by calculating the methemoglobin (metHb) quantity that forms after the same treatment (P < 0.00001). The statistical analyses we carried out showed that metHb formation was not affected by age, sex, smoking habits, red blood cell number, Hb, Ht or tumor staging. In this study, by using equal parameters of investigation, we noted that the blood of the subjects with ND who were previously treated with calcium-antagonists drugs and with antagonists of H1 and H2 receptors, gave results completely superimposable on the results obtained from healthy subjects, implying that the treatment had avoided the increase of FRs. Therefore we concluded that calcium antagonists and the antagonists of the H1 and H2 receptors behave as antioxidant substances, having decreased the FRs damaging activity on the cellular membranes, thus controlling, although to a limited degree, the pejorative evolution of the disease. It is also important to remember that investigations into the ND, even possible screenings, must take into account the above said data, submitting the subjects under investigation to a pharmacological wash out, particularly with those substances which, are considered to be scavengers of FRs. Some of these substances are investigated in this work. PMID- 8712696 TI - Control of renal carcinoma TC-1 growth, cyclin/kinase and nm23 expression by IFN gamma and TGF-beta. AB - To gain mechanistic insights into the growth control of renal cell carcinoma cells by IFN-gamma and TGF-beta, a recently established human renal carcinoma TC 1 cell line was treated with different concentrations of IFN-gamma and TGF-beta. Cell growth and changes in specific gene expression were evaluated. IFN-gamma exerted an antimitogenic effect on TC-1 cells, whereas TGF-beta was essentially without effect. The growth-suppressed cells had reduced expression of proliferating cell nuclear antigen (PCNA), the G2/M cell cycle transition regulatory proteins cyclin B/p34cdc2, the tumor suppressor gene pRB, and the antimetastatic gene nm23. However, levels of other cell cycle regulatory protein molecules such as cyclin D and p53 were unaffected by IFN-gamma. Thus, the antimitogenic effect of IFN-gamma may be mediated by its ability to modulate specific oncogene changes. PMID- 8712697 TI - Quantitative assessment of mixed hematopoietic chimerism by polymerase chain reaction after allogeneic BMT. AB - Patients who develop mixed hematopoietic chimerism (MHC) after allogeneic bone marrow transplantation (allo-BMT) might have an increased risk of relapse or graft failure. In order to identify patients with MHC we established a quantitative polymerase chain reaction (PCR) approach to estimate the individual degree of autologous recovery and investigate the dynamics of MHC posttransplant. Therefore standardized mixed chimeric samples were generated in each individual case by mixing pretransplant recipient and donor DNA at a range of percentages. After amplification the samples were analyzed densitometrically and signal intensities were taken as the basis of individual standard curves. Subsequently posttransplant DNA samples were also analyzed, the intensity of the informative signals were compared to the standard curves and autologous recovery was expressed in percentage related to host DNA. We investigated 40 pediatric patients receiving allo-BMT by this method. 10/40 developed MHC during course of the observation period. Each patient with MHC revealed a certain dynamic. This approach might be helpful when deciding on early intervention and additional treatment to prevent graft failure or relapse. PMID- 8712698 TI - Activated c-Ha-ras is not sufficient to produce the preneoplastic phenotype of human breast cell line MCF10AT. AB - MCF10AT cells are human breast epithelial cells which are able to establish preneoplastic lesions in immune deficient mice. Although, the preneoplastic phenotype was observed following transfection with a mutated c-Ha-ras (codon 12 valine), clones of MCF10AT are unable to form lesions in vivo. Restriction size fragment analysis was used to confirm that a clone unable to form the preneoplastic lesions retained the activated c-HA-ras and confirmed that the insertion site of the activated c-Ha-ras was the same for the clone as for MCF10AT1 which was selected for its ability to form lesions in vivo. Western blotting with antibody specific for the codon 12 valine c-Ha-ras demonstrated that p21 protein was comparable as well. Thus, the activated c-Ha-ras is not sufficient for the preneoplastic phenotype of human breast stem cell line MCF10AT. PMID- 8712699 TI - Combined effect of 5-fluorouracil and recombinant tumor necrosis factor against human gastric carcinoma cell lines. AB - This study was performed to investigate the direct effects of recombinant human tumor necrosis factor (rH-TNF) and recombinant human interleukin-2 (rH-IL-2), either alone or in combination, on the cytotoxicity of 5-FU measured by MTT assay against human gastric adenocarcinoma cell lines (MKN-28 and MKN-45), and also to determine the optimal schedule for their combination. The antitumor activity of rH-TNF was enhanced more than 42% by 10(2) U/ml of rH-IL-2. The enhancing effects of rH-TNF and rH-IL-2 on the cytotoxicity of 5-FU were evaluated in terms of Modification Index(MI), the MI value at 10 U/ml rH-TNF was 1.6; the MI at the same concentration of rH-TNF in the presence of 10(2) U/ml of rH-IL-2 was 2.1. These results demonstrated that the antitumor effect of 5-FU was enhanced 1.6 times by 10 U/ml of rH-TNF and further enhanced by the combined use of rH-TNF and rH-IL-2. The combined effect of equal concentrations of 5-FU and rH-TNF was superior or equivalent to that of 5-FU or rH-TNF alone. The sequence of 5-FU followed by rH-TNF and rH-IL-2 showed a higher inhibitory effect than the reverse sequence. This sequence combination seems worthy of further consideration for the treatment of gastric cancer. PMID- 8712700 TI - Cell activation and death (apoptosis) induced by IL-2: ultrastructural evidence. AB - Peripheral blood cells of cancer patients with advanced renal carcinoma and treated with interleukin-2 by intravenous application were studied by electron microscopy at different intervals from the beginning of rIL-2 administration. Morphofunctional modifications of lymphocytes, circulating hystiocytes containing phagocitated bodies and polimorphonucleated cells undergoing phagocytosis were observed with a time dependent increase of altered cells. The analysis of blood cells cultured in the presence of recombinant IL-2 confirmed the in vivo results. Our data suggest that IL-2 induces apoptotic phenomena in the peripheral blood cells of treated patients. PMID- 8712701 TI - Effects of glycosylated and non-glycosylated G-CSFs, alone and in combination with other cytokines, on the growth of human progenitor cells. AB - Two recombinant human granulocyte colony-stimulating factors (rHu G-CSF) are clinically available, a glycosylated (lenograstim) and a nonglycosylated from (filgrastim). Since there is accumulating evidence that glycosylation plays a role in the in vitro activity of the G-CSF molecule, we compared the biological potency of lenograstim and filgrastim on human hematopoietic progenitor cells by colony assay in semisolid medium and ex vivo expansion experiments. Leukapheretic products without further processing and CD34-positive purified cells were used as source of human progenitors. Lenograstim demonstrated greater capacity, to stimulate the colony growth of both, purified and CD34+ peripheral blood cells. This effect, which is evident especially at low doses of growth factor, seems not to be mediated by accessory cells. Whether these observations may have clinical relevance is still to be clearly assessed and further investigations are needed. PMID- 8712702 TI - Non-random primary and secondary chromosomal abnormalities in human gastric cancers. AB - By using Giemsa-banding and fluorescence in situ hybridization techniques, we have been able to identify primary and secondary cytogenetic abnormalities in four gastric tumors at different stages of development. Structural and numerical abnormalities were present in all four gastric tumors in chromosomes 3, 7, 11, and X. Other abnormalities involving chromosomes 1, 5, 6, 8, 13, 15, 17, 18, 19 and 22 were observed, but only in three advanced gastric tumors, suggesting that these were secondary/tertiary genetic defects. Based on these results it was possible for us to decipher primary and secondary genetic abnormalities in these four gastric tumors. PMID- 8712703 TI - Characterization of two urothelium cancer cell lines derived from a blackfoot disease endemic area in Taiwan. AB - Established cancer cell lines with distinct characteristics are valuable in many aspects of in vitro study pertaining to tumor biology, as well as the possible geographic significance. Two long-term cell lines, BFTC905 and BFTC909, were established from primary transitional cell carcinomas (TCC) of patients from a blackfoot disease endemic area in Taiwan, where the prevalence and standard mortality rates of this cancer are unusually high. BFTC905 derived from a grade III TCC of urinary bladder has an epitheloid morphology, and BFTC909 derived from the sarcomatoid component of a grade III TCC of renal pelvis exhibits a fibroblastic growth pattern. They share similar morphological and immunocytochemical characteristics with the tumors of origin, have multiple chromosomal aberrations, and are tumorigenic in nude mice. BFTC905 reveals a unique homogeneously staining region at 11q13, the amplification of which has been detected in 20% of transitional cell carcinoma. In addition to the rarity of renal pelvic origin, the vimentin expression associated with an aggressive clinical behavior and a relatively high tumorigenicity as demonstrated by BFTC909 cells has also rarely been mentioned in TCC cell lines, but has been well documented in vivo and in vitro in renal cell carcinoma and breast cancer. PMID- 8712704 TI - Prostate-specific antigen promoter driven gene therapy targeting DNA polymerase alpha and topoisomerase II alpha in prostate cancer. AB - Attainment of cell type-specific cytotoxicity with minimal side effects is the ultimate goal of cancer therapy. By employing the prostate-specific antigen promoter (PSAP), we investigated (1) whether PSAP-driven antisense genetic constructs targeting DNA polymerase-alpha and topoisomerase II alpha (Top II alpha), designated PSAP-antipol and PSAP-antitop respectively, could induce death of prostate cancer cells, and (2) whether the cytotoxicity is restricted to cells of prostate origin. A PSAP-driven beta-galactosidase gene, PSAP-LacZ, was also used to estimate the expression of the PSAP-driven transcripts. Lipofection mediated gene transfers were performed with these 3 constructs and a control plasmid, pCDNA3, in 3 human prostate cancer cell lines (LNCaP, DU-145, PC-3) and 5 other cell lines (Cos-1 [monkey kidney], HL-60 [human myeloid leukemia], Hep G2 [human hepatoma], NCI H460 [human lung cancer] and SW 480 [human colon cancer]). On transfection with PSAP-LacZ, LNCaP, DU-145, and PC-3 showed a 10.8, 1.8, and 1.6 fold increase in beta-galactosidase activity, respectively. The remaining 5 cell lines showed no changes after transfection. Corresponding to the levels of the induced beta-galactosidase activity, LNCaP showed the strongest growth inhibition by the antisense constructs: 36% by PSAP-antipol, 39% by PSAP-antitop and 80% by PSAP-antipol+PSAP-antitop. DU-145 and PC-3 had minimal growth inhibition with PSAP-antipol alone or PSAP-antitop alone. However, when cotransfected with PSAP-antipol and PSAP-antitop, DU-145 and PC-3 displayed 42% and 55% growth inhibition, respectively. In contrast, no cytotoxicity was observed in the remaining 5 cell lines when transfected with PSAP-antipol, PSAP antitop or both. Therefore, PSAP-driven antisense gene therapy targeting DNA polymerase-alpha and Top II alpha inhibits the growth of human prostate cancer cells and the cytotoxic effect is restricted in cells of prostate origin. PMID- 8712705 TI - Synthesis of ether-linked analogues of lysophosphatidate and their effect on the proliferation of human epithelial cancer cells in vitro. AB - To investigate whether lysophosphatidate analogues of alkyllysophospholipids were antiproliferative we synthesized three new ether-linked analogues of lysophosphatidic acid and investigated their antiproliferative activity on epithelial cancer cell lines derived from different tissues. The antiproliferative effects of the compounds on MCF-7 and T47D (breast), A549 and A427 (lung), A498 (kidney), SK-N-SH and SK-N-MC (neuroblastoma), and DU145 (prostate) cells were compared with the ability of 1-O-octadecyl-2-O-methyl glycero-3-phosphocholine, the archetypic alkyllysophospholipid, to inhibit the proliferation of all the cell lines. 1-O-Hexadecyl-2-O-methyl-sn-glycero-3 phosphate and 4-thiohexadecyl-3(S)-O-methoxybutane-4-phosphate were unable to inhibit the proliferation of any of the cells to any degree, while slightly enhancing the proliferation of DU145 cells. In contrast 4-O-hexadecyl-3(S)-O methoxybutanephosphonate was a potent antiproliferative agent that was on the whole more active than 1-O-octadecyl-2-O-methyl-glycero-3-phosphocholine. Since 1 Oleoyl-2-lyso-phosphatidate (LPA) was non-mitogenic in all the cell lines except the neuroblastoma line SK-N-SH, it is unlikely that the inhibition of cell proliferation by 4-O-hexadecyl-3(S)-O-methoxybutanephosphonate was a consequence of perturbation of cellular response to the mitogenic effects of LPA. PMID- 8712706 TI - Estramustine-binding protein (EMBP) content in four different cell lines and its correlation to estramustine induced metaphase arrest. AB - It is known that estramustine (EM) accumulates in cells at the G2/M-phase and causes metaphase arrest of various cell types. The inhibitory effect is mediated by interaction with microtubule-associated proteins (MAPs) and/or tubulin. Estramustine-binding protein (EMBP) is a secretory protein which has been found in a number of different tumor cells and has been shown to faciliate the uptake of EM into cells. In this study the efficacy of EM in arresting cells at metaphase was studied, using four different human cell lines; the prostatic cancer cell line DU 145, the breast cancer cell line MDA 231, the colon cancer cell line Colon 320, and the urinary bladder cancer cell line RT4. The cells were incubated with EM at a concentration of 10 micrograms/ml for 24 hours. The data reveal an increase in metaphase arrests in the DU 145 and in Colon 320 cell lines. Both of these cell lines were found to contain high amounts of EMBP using a dot-blot assay. The other two cell lines, MDA 231 and RT4 had undetectable intracellular amounts of the protein and exhibited a low increase in metaphase arrests. The cell lines were analysed regarding S-phase fraction with flow cytometry (FCM) to exclude the growth rate of the cells as a limiting factor. The results from the FCM confirmed the cytogenic analysis, that is a higher percentage of cells were in the G2/M phase in both the DU 145 and Colon 320 cell line compared to MDA 231 and RT4. EM causes mitotic arrest in those cell lines that contain detectable amounts of EMBP. PMID- 8712707 TI - Growth-suppressive activities of serine protease inhibitors, ONO-3403 and ONO 5046, toward normal and transformed murine fibroblasts. AB - ONO-3403 and ONO-5046 are potent synthetic protease inhibitors of trypsin and elastase, respectively. These compounds suppressed the proliferation of polyoma virus- and Kirsten sarcoma virus-transformed BALB/c 3T3 cells more effectively than their normal counterparts. SV40-transformed 3Y1 and v-Ha-ras-transformed NIH3T3 cells were also more sensitive to ONO-3403 and ONO-5046 than the parent normal cells. These results suggest that ONO-3403 and ONO-5046 are useful for selective suppression of the proliferation of rapidly growing transformed cells. PMID- 8712709 TI - Antitumor activity of an alkylating derivative of dipyridamole. AB - Synthesis of 2,6-Bis[bis(2-chloroethyl)amino]-4,8-dipiperidino-pyrimido [5,4 d]pyrimidine (DIP-C1) was carried out, and the new derivative showed cytotoxic activity comparable to other alkylating drugs on cultured P388 leukaemia cells and HeLa cells. The present paper reports the effects of DIP-C1 on respiration of Ehrlich ascites tumor cells and on survival of the mice implanted with Ehrlich ascites tumor cells. The compound showed a significant activity in both experimental models. PMID- 8712708 TI - Effects of coffee cherry on the immune system in SHN mice. AB - Previously, we found that chronic ingestion of the extract of coffee cherry (CC), the residue after the removal of coffee beans, induced a marked suppression of the development and the growth of spontaneous mammary tumours in a high mammary tumour strain of SHN mice. As a possible step to clarify the mechanism of this effect, the immunomodulating role of CC was examined in this study. CC treatment resulted in significant weight gain in the spleen. CC treated experimental mice showed a significant decrease in the proportion of immature (CD4+8+) thymocytes and an increase in the percentages of mature cells expressing helper/inducer (CD4+8-) or cytotoxic/suppressor (CD4-8+) phenotypes. The proportion of T cells expressing CD25, a lymphocyte activation marker, in the spleen and peripheral blood tended to increase in the CC treated group. The natural killer activity of the spleen cells was not affected by CC ingestion. These results have revealed that CC can enhance the differentiation of thymocytes and the activation of peripheral T lymphocytes. PMID- 8712710 TI - BK virus-induced osteosarcoma (Os515) as a model of human osteosarcoma. AB - BK virus was isolated by Gardner et al in 1971 from the urine of an immunosuppressed patient with a kidney allograft. Antibodies to this virus are ubiquitous in the general populations worldwide, but the oncogenic capacity of BKV in humans had not been reported. The virus transformed in vitro permissive human and non-permissive animal cells, and the transformed cells had the T antigen. Intracerebral and intravenous inoculation of BKV in newborn hamsters induced malignant tumours (mainly ependymomas, malignant insulinomas, and osteosarcomas). Subcutaneous and intraperitoneal routes were also effective. The virus was only rescued from a few tumours by fusion with human embryonic cells or Vero cells. Brain tumours appeared earlier and osteosarcomas developed in animals which survived for more than 6 months. Many of the osteosarcomas were bony and grew slowly with frequent lung metastases, and a few osteosarcomas were soft and grew rapidly without lung metastases. Experimental targeting chemotherapy with doxorubicin (DX)-containing immunoliposomes was performed against Os515 osteosarcoma. In in vitro experiments, DX-Lip-MoAb29 showed a more significant inhibitory effect on cultured Os515 cells than free Dx and DX-Lip. DX-Lip DNP had less effect. In in vivo experiments, DX-Lip-MoAb29 suppressed the growth of Os515 tumour isografts in hamsters and prolonged the survival of recipients more significantly than free DX. PMID- 8712711 TI - Growth control of human prostatic cancer cells by the phorbol ester TPA: possible involvement of protein kinases. AB - To gain insights into the involvement of kinases in the growth control of prostate carcinoma cells by the phorbol ester 12-O-tetradecanoylphorbol-13 acetate (TPA), immunoblot analysis and in vitro phosphorylation assays were performed using kinase discriminating effectors and extracts prepared from the androgen-dependent LNCaP and the androgen-independent JCA-1 human prostate cells. The down-regulation of PKC-alpha and -beta in JCA-1 cells was correlated with the effects of TPA. In LNCaP cells, proliferation may involve DNA-PK, proposed to act possibly via phosphorylation of the androgen receptor. PMID- 8712712 TI - The activity of N-(hydroxymethyl) melamines in fresh human ovarian tumour cells and xenografts. AB - Trimelamol (TM) was developed as a water soluble analogue of the oral chemotherapeutic agent hexamethyl-melamine (HMM), to be administered i.v., in an effort to avoid dose limiting emesis. Because of formulation difficulties due to its inherent instability the development of TM was halted. In vivo studies using a human ovarian cancer xenograft model PXN/65 showed TM to be curative in the dose range of 15-60 mg/kg i.p. daily x 5, for 4 weeks. Conversely, HMM given at the highest dose (60 mg/kg i.p., or 90 mg/kg p.o.) indicated only very modest tumour growth delays. In vitro chemosensitivity testing using primary ovarian tumour cultures showed that in 12/23 cases indicating reduced sensitivity to cisplatin or carboplatin, sensitivity to TM was increased. TM was curative in the carboplatin-resistant HX 110P human ovarian cancer xenograft and promising activity was seen in the MX-1 human breast cancer xenograft. In spite of enhanced stability in aqueous solution and good in vitro cytotoxicity, the TM analogues CB 7669 (triscyanomethyl) and CB 7639 (tristrifluoroethyl) showed disappointing in vivo antitumour activity which may be explained by the need for prolonged exposure. TM analogues with intermediate stability are currently under development in an effort to further the clinical development of this promising group of antitumour agents. PMID- 8712713 TI - Metabolism, protein binding and in vivo activity of the oral platinum drug JM216 and its biotransformation products. AB - This study evaluates the metabolism of the oral platinum drug JM216 [bis(acetato) amminedichloro (cyclohexylamine)platinum (IV)] following oral administration to Balb C- mice. JM216 was detectable 1 h post administration in mice but not in patients. Also, a late eluting metabolite observed in patients was not detected in mice. JM118 [amminedichloro(cyclohexylamine) platinum (II)], the platinum II species which is the major metabolite in patients was rapidly converted following i.v. administration to a compound having the same retention time as JM383 [bis(acetato)ammine(cyclohexylamine)dihydroxo platinum(IV)] indicating that the levels of JM383 following JM216 administration have probably been overestimated. The metabolite D observed in patients for which a structure has not been assigned, was also detected in mice. However, it did not originate from any of the identified biotransformation products. The protein binding evaluated in plasma, and buffer with physiological levels of albumin and globulin showed that only Platinum (II) species have significant binding and that Jm118 showed the same affinity to albumin and globulin (t 1/2 of 4.2 and 4.8 h) while cisplatin bound more readily to albumin (t 1/2 3.4 h) than globulin IV (t 1/2 8.2 h). JM216 itself failed to bind to either of the proteins tested indicating extensive reduction in patients, animals or plasma incubation medium. JM118 and JM518 [bis(acetato)amminechloro(cyclohexylamine) hydroxoplatinum (IV)] were significantly more active than the platinum IV complexes JM216 and JM383 when given i.p. to ADJ/PC6 plasmacytoma bearing mice (ED90 of 1.0 and 0.4 versus 5.7 and 4.2 mg/kg, TI (therapeutic index) of 14 and 37 versus 5.3 and 4.2). When given orally, JM216 was the most potent drug (ED90 of 5.8 versus 11,12 and 42 mg/kg and TI of 57 versus 12 12 and 16) for JM118 and JM383. There data indicates that JM216 biotransformation products are potent but that the levels of JM383 determined in our analytical conditions could have been overestimated. PMID- 8712714 TI - Experimental colonic carcinogenesis: a new animal model. AB - The aim of this work was to develop an animal model of accessible colonic tumours simulating human carcinogenesis. A segment of the colon was exteriorized to the skin in 60 rats and swabbed with N-methyl-N-nitrosourea. All the animals developed tumoral abnormalities or frank dysplasic tissue 7 months after the beginning of the experiment. This animal model which closely mimics human colonic carcinogenesis is particularly interesting because tumour development can be monitored in live animals without interrupting normal intestinal functions. PMID- 8712715 TI - Human DNA topoisomerase II: biochemistry and role in chemotherapy resistance (review). AB - Recently, it has been discovered that DNA topoisomerases are the target of many anti-cancer drugs which were already widely used in the clinic. Using the latest molecular biological techniques much has been learned about the function of DNA topoisomerases in normal cells and in cancer cells, and about how anticancer drugs inhibit these enzymes. In this review we present an overview of the function of human type II DNA topoisomerases, how they are inhibited by certain drugs and on how cancer cells may become resistant to these drugs. PMID- 8712716 TI - Discrimination between tumour and normal cells by staining with 3,4,5,6,16,17 hexadehydro-16-(methoxycarbonyl)-19 alpha-methyl-20 alpha-oxayohimbanium: the uracil ring as a target for the specific interaction between RNA(s) and the fluorescent probe. AB - 3,4,5,6,16,17-Hexadehydro-16-(methoxycarbonyl)-19 alpha-methyl-20 alpha oxayohimbanium (Alstonine) is a fluorescent alcaloid which has been known to stain tumour cells more efficiently than normal ones. In this paper the spectral properties of Alstonine were first investigated and its capability for preferential staining of tumour cells verified in culture using SK-OV-3 cells as tumour cells and Mouse 3T3 fibroblasts as controls. Then interactions between Alstonine and biological macromolecules were investigated to provide the rationale for preferential labelling. Molecular filtration techniques have demonstrated that binding occurs only with RNA molecules. Similar experiments were performed with different isopolynucleotides to find an explanation for that specificity. They provide evidence that binding occurs only in the presence of a uridyl ring. This is consistent with the specificity of the linkage to RNA. As the linkage of Alstonine with RNA did not induce any shift or obvious change in the intensity of its fluorescence spectrum, it is concluded that the binding might involve the side chain of the fluorescent compound. PMID- 8712717 TI - Evidence for different mechanisms of EMT-6 tumor necrosis by photodynamic therapy with disulfonated aluminum phthalocyanine or photofrin: tumor cell survival and blood flow. AB - A comparison was made of photodynamic therapy (PDT) mediated by two photosensitizers, the disulfonated aluminum phthalocyanine (AlPcS2) and Photofrin* (PII) with regard to their mechanism of action on murine tumors. Balb/c mice bearing intradermally growing EMT-6 tumors were injected intravenously with either 1 mumol kg-1 body weight of AlPcS2 or 5 mg/kg of PII 24 h prior to red light irradiation from a Xenon lamp (650-700 nm, 200 mW cm-2, for AlPcS2 and 600-650 nm, 400 J cm-2 for PII. Tumor cell survival following in vivo PDT was determined by an in vitro clonogenicity assay on the dissociated tumors. Immediately after the completion of light irradiation, a reduction of approximately 72% in the number of clonogenic cells was seen with AlPcS2-treated tumor versus approximately 24% of that for PII-treated tumor. Further loss of clonogenic cell survival progressed as a function of time following PDT, and was considered to be the consequence of indirect PDT action, however, the decline in cell viability was steeper in the first 6 h with PII-PDT than with AlPcS2-PDT. 24 h after PDT, the clonogenic capacity of both AlPcS2-and PII-PDT treated tumor fell to approximately 3% of the control tumor. The PDT effect on tumor blood flow as a measure of the tumor vascular damage was monitored by the retention of 99mTc MIBI in the tumor. Little effect on tumor blood flow was seen with AlPcS2-PDT at 0 h after the completion of light treatment. Thereafter the blood flow declined slowly and remained at approximately 50% the level of the control by 24 h post PDT. In contrast, PII provoked a approximately 40% reduction of tumor blood flow immediately after the completion of photo irradiation, which then fell to approximately 20% within 2 h and approximately 7% by 24 h post-PDT. These results indicate the involvement of both direct and indirect mechanisms in the PDT induced tumor necrosis. However, AlPcS2-PDT exerted a larger direct tumor cell phototoxic effect, whereas PII-PDT induced tumor cell death to a greater extent via an indirect effect that parallels the extensive damage to the tumor vasculature. PMID- 8712718 TI - Relative efficacy of the opioid antagonist, naltrexone, on the initiation and promotion phases of rat mammary carcinogenesis. AB - We have shown earlier that naltrexone, a long acting opioid antagonist, inhibits the development and growth of 7,12-dimethylbenz(a)anthracene (DMBA)-induced rat mammary tumors. We extended these studies to determine the independent effects of naltrexone when fed at 75 mg/kg diet during the initiation (I), promotion (P), or initiation plus promotion (I+P) phases of DMBA-induced rat mammary carcinogenesis. The percentage of rats with palpable tumors as well as the number of tumors per rat were determined during the 130 day experiment. When fed during the 1, P, or I+P phases, mammary tumor incidence was significantly inhibited by 27%, 60% and 45% respectively, as compared to the control group. Similarly, tumor multiplicity was significantly reduced by naltrexone treatment. Tumor multiplicity was reduced by 40%, 73%, and 70% when fed naltrexone during I, P, and I+P respectively, as compared to the controls. These results indicate that naltrexone acts on the I and P phases, with the effects being maximal when fed during the P and I+P phases. PMID- 8712719 TI - A small RNA is associated with dermatan sulfate proteoglycan. AB - A specific RNA, called pgRNA, was bound to purified dermatan sulfate proteoglycan from porcine skin. The pgRNA was approximately 20 bases in length and contained greater than 80% guanosine. The pgRNA-proteoglycan complex was dissociated into pgRNA and apo-proteoglycan by denaturing conditions. The complex was reconstituted from apo-proteoglycan and pgRNA, but not by other RNA molecules. Both pgRNA and an additional smaller RNA, perhaps derived from pgRNA, were isolated complexed to the glycosaminoglycan dermatan sulfate derived from dermatan sulfate proteoglycan. Both the chromatographic and physical properties of the dermatan sulfate were influenced by the pgRNA binding. PMID- 8712721 TI - Interferons inhibit EGF-stimulated cell growth and reduce EGF binding in human breast cancer cells. AB - CG-5 estrogen-sensitive human breast cancer cells contain specific Epidermal Growth Factor receptors (EGF-R, Kd = 0.09-0.17 nM) and respond to the mitogenic effect of EGF. The increase in cell proliferation has been observed starting with very low concentrations of EGF (10-12M) and was statistically significant at all doses. Nevertheless, cell growth stimulation was emphasized when cells were grown under stringent culture conditions. When cells were exposed to 100 IU/ml of natural beta-interferon (n beta-IFN) the binding of EGF to the cell membrane was reduced after 72 hours of treatment, while the exposure of CG-5 cells to 1000 IU/ml of n beta-IFN resulted in an EGF-R reduction which started after 48 hours and became statistically significant after 72-120 hours. If CG-5 cells were treated with 1000 IU/ml of recombinant alpha 2b-interferon (ra2b-IFN) this reduction was observed after 168 hours of exposure to the drug. Both the IFNs abolished EGF-stimulated cell growth. Our results indicate that IFN treatment down-regulates EGF-R in estrogen-sensitive breast cancer cells and suggests that this down-regulation may be involved in the inhibitory action of IFN on cell growth. PMID- 8712720 TI - Antitumor activity of phenothiazine-related compounds. AB - One of the biggest challenges in health care is the fight against tumors. Some phenothiazines have antitumor activity on HEp-2 tumor cells. In this study, we tested the antitumor effects of three series such as 10-nonsubstituted phenothiazines, 10-[n-(phthalimido)alkyl]-2-substituted-10H-phenothiazines and 1 (chloroethyl)-3-(2-substituted-10H-phenothiazines-10-yl)alkyl-1-ureas with H, Cl and CF3 substitution at position C2. The TCID50 of phenothiazines was affected by the H, Cl and CF3 at C2. Trifluoromethyl derivative of phenothiazine showed potent (R = CF3, TCID50 = 4.7 micrograms) activity, whereas the chlorine derivative of phenothiazine (R = Cl, TCID50 = 62.5 micrograms) had a relatively weak effect. In the group of 10-[n-(phthalimido)alkyl]-2-substituted-10H phenothiazines, 10-[3-(phthalimido)propyl]-10H-phenothiazine (R = H, n = 3, TCID50 = 11.5 micrograms), 10-[4-(phthalimido)butyl]-10H-phenothiazine (R = H, n = 4, TCID50 = 7.8 micrograms) and 10-[3-(phthalimido)propyl]-2-trifluoromethyl 10H- phenothiazine (R = CF3, n = 3, TCID50 = 11.5 micrograms) was very effective. On the other hand, TCID50 of 10-[3-(phthalimido)propyl]-2-chloro-10H phenothiazine (R = Cl, n = 3, TCID50 = 75.0 micrograms), 10-[4 (phthalimido)butyl]-2-chloro-10H-phenothiazine (R = Cl, n = 4, TCID50 = 31.3 micrograms) and 10-[4-(phthalimido)butyl]-2-trifluoromethyl-10H-phenothiazine (R = CF3, n = 4, TCID50 = 50.0 micrograms) were about 4-8 times less effective than 10-[4-(phthalimido)butyl]-10H-phenothiazine (R = H, n = 4, TCID50 = 7.8 micrograms). Among six 1-(chloroethyl)-3- (2-substituted-10H-phenothiazin-10 yl)alkyl-1-ureas, two chlorine compounds such as 1-(2-chloroethyl)-3-(2-chloro 10H-phenothiazin-10-yl)propyl-1-urea (R = Cl, n = 3, TCID50 = 6.3 micrograms), 1 (2-chloroethyl)-3-(2-chloro-10H-phenothiazin-10-yl) butyl-1-urea (R = Cl, n = 4, TCID50 = 7.8 micrograms), and 1-(2-chloroethyl)-3-(2-trifluoromethyl-10H phenothiazin-10-yl)buty l-1-ur ea (R = CF3, n = 4, TCID50 = 7.8 micrograms) were significantly active. Tests showed that the substitution at 2C position apparently affected the anti-HEp-2 tumor cell activity; that the length of the aliphatic side chain at 10N contributes to the anti-tumor activity; and that the TCID50 values of the derivatives with butylene group (-C4H8-) were lower than those with propylene group (-C3H6-) except 10-[4-(phthalimido) butyl]-2 trifuoromethyl-10H-phenothiazine and 1-(2-chloroethyl)-3-(2-chloro-10H-pheno thiazin-10-yl) butyl-1-urea. PMID- 8712722 TI - In vitro chemosensitivity testing of human tumours by collagen gel droplet culture and image analysis. AB - It is necessary to develop an in vitro test to overcome the problems often associated with in vitro chemosensitivity tests on individual human tumours. We have developed a collagen gel droplet culture technique that allows for a three dimensional in vitro growth and drug response assay for human solid tumour cells. Important features of chemosensitivity testing by the collagen gel droplet culture technique include the maintenance of high cloning efficiencies resulting in the need for fewer tumour cells, sufficient suppression of the in vitro proliferation of contaminating non-malignant cells by serum-free medium, and the application of the image analysis system which automatically discriminates between cancer cell colonies and fibroblasts. We described in vitro-in vivo correlations for drug response using 7 human lung cancer xenografts grown in the collagen gel droplet culture and as xenografts in nude mice. Results demonstrated significant correlations with the in vitro drug concentration at 1/10 of the peak plasma concentrations (1/10 Cmax) with the correlation coefficient 0.84 for all four drugs tested. We have cultured 206 tumours thus far obtaining 86% of evaluability for drug response. The drug response data of the fresh lung cancers were similar to not only to data for lung cancer lines but the reported clinical pattern. These results suggested that the collagen gel droplet culture at 1/10 Cmax may have potential in predicting clinical drug responses. PMID- 8712723 TI - Expression of CDK inhibitor genes in immortalized and carcinoma derived breast cell lines. AB - The expression of the CDK inhibitor (CDI) genes p15(INK4B), p16(INK4A), p18 and p21Cip1 was examined in immortalized, non-tumorigenic cell lines derived from human breast epithelium, and in breast carcinoma derived lines. An increase in p16 expression, suggesting loss of pRb function, was recorded in two immortalized lines, and complete absence of p16 mRNA was observed in the third. In contrast, high levels of p21Cip1 mRNA were found in two immortalized lines. In addition to differences in p16 and p21Cipl, variations in the expression of p15 and p18 mRNA were observed between different cell lines. Immortalized A1N4 and HBL100 cells, as well as ER+, MCF-7 carcinoma cells, expressed high levels of p15 mRNA. A1N4, HBL100 cells and highly malignant ER MDA-MB-231 cells expressed high levels of p18 mRNA. Inhibition by genistein indicated that p18 mRNA expression was dependent on cellular tyrosine kinases in these cells. We conclude that the pattern of p15 and p18 mRNA expression was distinct from that of p16 and p21Cip1, suggesting different modes of regulation. PMID- 8712724 TI - Effect of the type of serum in the medium on sodium ascorbate-induced cytotoxicity. AB - Sodium ascorbate induced cytotoxicity against human glioblastoma T98G cells in RPMI1640 medium supplemented with fetal bovine serum or human serum samples was studied. Several human serum samples significantly reduced the cytotoxic activity of sodium ascorbate, regardless of sex, age or the disease of the serum donor with or without heat-inactivation of the serum. ESR spectroscopy revealed that this serum effect was not simply due to the alteration of the ascorbyl radical intensity, produced from sodium ascorbate. The present study suggests that the apoptosis-inducing activity of sodium ascorbate might be significantly affected by human serum. PMID- 8712725 TI - Induction of programmed cell death (apoptosis) in human lymphoid leukemia cells by catechin compounds. AB - The need for antitumor compounds with novel mechanisms of action is great. The exposure of human lymphoid leukemia Molt 4B cells to epigallocatechin (EGC), epigallocatechin gallate (EGCG), and persimmon extract (PS) led to both growth inhibition and the induction of programmed cell death (apoptosis). The fragmentation of DNA to oligonucleosomal-sized fragments, characteristic of programmed cell death, was determined to be concentration- and time-dependent. These data provide the first evidence that catechin compounds and persimmon extract containing all these catechin compounds induce programmed cell death. PMID- 8712726 TI - Overexpression of p53 protein in benign and malignant laryngeal epithelial lesions. AB - This study investigated the immunohistochemical expression of p53 protein in various laryngeal lesions using Kambic's classification from simple hyperplasia to invasive squamous cell carcinoma (SCC), and analyzed the relationship between p53 protein overexpression and the severity of epithelial abnormalities. p53 overexpression was observed in 10/19 (53%), in 9/16 (56%), and in 9/13 (69%) cases of simple, abnormal, and atypical hyperplasia respectively, and also in 8/10 (80%) cases of SCC. The proportion of immunoreactive cells and staining intensity increased with the progression of the epithelial hyperplastic lesions. Our study confirms the association of p53 protein overexpression with laryngeal epithelial hyperplastic lesions which have the potential to transform into malignancy, but considering the follow-up of the patients, p53 expression cannot be considered a reliable prognostic factor for any group of laryngeal epithelial hyperplastic lesions regardless of the severity of the lesions. PMID- 8712727 TI - Modifications of interphasic NORs as a diagnostic parameter of atypical lesions of the female breast. AB - Seventy-five breast samples including normal tissue, hyperplastic, metaplastic, atypical and neoplastic lesions were employed for the determination of interphasic Nucleolar Organizer Regions (NORs) modifications and Proliferating Cell Nuclear Antigen (PCNA) immunoreactivity. Interphase NORs were quantitatively and qualitatively modified in atypical lesions and breast carcinomas, whereas only modifications in the Ag-NORs count were found in benign samples. Our results investigated the nature of interphase NORs in the hope of finding a use for their evaluation in the diagnosis and biological clarification of breast epithelial atypia. PMID- 8712728 TI - Effects of c-myc oncogene modulation on drug resistance in human small cell lung carcinoma cell lines. AB - Small cell lung carcinoma (SCLC) is characterized by rapid development of resistance to drugs, such as cisdiamminedichloroplatinum (II) (cDDP) and anthracyclines. The molecular basis for resistance to cDDP and adriamycin (Adr) is poorly understood. One of the genetic alterations observed in SCLC, which is correlated with poor prognosis, is amplification and overexpression of c-myc oncogene. Therefore, activation of the c-myc oncogene might form a basis for resistance. The relationship between c-myc and cDDP as well as Adr resistance was analyzed by down-regulation of endogenously expressed c-myc in the human SCLC cell line GLC4, and its cDDP and Adr resistant sublines (GLC4-cDDP and GLC4-Adr). Cells were incubated with an unmodified antisense (AS) oligodeoxynucleotide complementary to the first 5 codons of the c-myc mRNA in serum free culture medium. Pre-incubation with 15 microM AS c-myc, reduced c-myc protein expression and induced 30-35% growth inhibition in all 3 cell lines. It resulted in increased cDDP sensitivity in GLC4-cDDP but not in GLC4. However, this pre incubation did not affect Adr sensitivity in all lines. The effect of AS c-myc pretreatment on cDDP resistance was not mediated by changes in cell cycle distribution. These findings suggest that c-myc plays a role in cDDP resistance by effects other than those on cell cycle distribution. PMID- 8712729 TI - Involvement of macrophages and cytokines into rejection mechanism of the drug resistant and immunogenic murine lymphoma P388/adria. AB - Macrophages and their products may exert either inhibitory or stimulatory effects on malignant cells,thus preventing or supporting tumor growth, however, the mechanisms of this interaction are not fully understood. It was the aim of the present study to elucidate the role of macrophage activation during the growth and rejection of highly immunogenic murine leukemia P388/adria cell line which was made resistant by suboptimal treatment of mice with adriablastin during the serial passaging of parental P388 cells. The functional activity of peritoneal macrophages and the serum level of cytokines IL-1 beta, IL-6 and TNF-alpha were studied in different groups of mice. Mice from group 1 (control) received saline. Mice from group 2 (tumor bearers) with fast subcutaneous (s.c) 100% tumor growth were compared with animals from group 3 that had been twice previously immunized with lethally irradiated P388/adria cells and later inoculated with viable tumor cells. Tumors grew in only 25% of group 3 animals with a significant delay. The activity of peritoneal macrophages was studied by NO2- production and the NBT test. Both tests revealed the early high systemic activation of macrophages in group 2. This coincided with the elevation of serum TNF-alpha and IL-6 levels. This effect was not dependent on whether alive or lethally irradiated tumor cells were inoculated. The NO2- production by peritoneal macrophages correlated well with the dynamics of serum cytokine levels while the NBT-test did not. Studies on group 3 showed total abrogation of early macrophage and cytokine reactions. The production of inhibitory factors by macrophages in previously immunized mice is suggested. The fact that the early activation of macrophages and increase of serum levels of proinflammatory cytokines occurred in animals with fast growing tumors, which was decreased or absent in animals with tumor delay or rejections, allows us to suppose that this reaction plays more a supporting than a protecting role for tumor growth. PMID- 8712730 TI - Apoptosis in human breast and gastrointestinal carcinomas. Detection in histological sections with monoclonal antibody to single-stranded DNA. AB - We report application of a novel immunohistochemical procedure for the staining of apoptotic (AP) cells in paraffin sections using monoclonal antibody (MAb) to single-stranded DNA. MAb differentiated between apoptosis and necrosis and in contrast to in situ end labelling specifically stained only AP cells. AP carcinoma cells stained with the antibody were detected in 32 of 58 infiltrating human breast carcinomas and in 9 of 15 colon adenocarcinomas. Stromal cells stained with the MAb were observed in all carcinomas, including those in which no AP carcinoma cells were detected. There was a strong positive correlation between the presence of AP cells, loss of hormone receptors and a high proliferation rate in breast carcinomas. AP cells were present in 80-87% of receptor-negative carcinomas, while most of receptor-positive breast carcinomas did not contain AP cells. Apoptosis in tumor cells was detected significantly more frequently among breast carcinomas with high, than among carcinomas with low S-phase fraction. AP cells were present in 93-95% of breast carcinomas which were receptor-negative and had a high S-phase fraction. Immunostaining demonstrated a strong positive correlation between the loss of bcl-2 protein and intensive apoptosis in breast carcinomas. Association between apoptosis and markers of poor prognosis in breast cancer (loss of hormone receptors, intensive proliferation, loss of bcl-2 protein) indicates that apoptotic cell death is typical of more aggressive carcinomas. PMID- 8712731 TI - Inherited predisposition to breast carcinoma. Results of first round examination of 537 women at risk. AB - Five hundred and thirty seven women at risk for breast carcinoma were identified. Family history was detailed and each woman given genetic counselling. Diagnostic examination for breast carcinoma was performed at the major hospitals of Norway, and included physical examination by expert surgeon, mammography and/or ultrasonography, and fine needle aspiration cytology when appropriate. Altogether 8 carcinomas and 5 cases of atypical hyperplasia were found, compared with 1.6 and 0.3 expected, respectively, from population studies. The finding exceeded the expected numbers described by autosomal dominant inheritance. In addition we found one carcinoma in situ. It is concluded that the methods employed are suitable to identify and examine women at risk for breast carcinoma. It is suggested that atypical hyperplasia may be the precancerous lesion, and should be treated as such. PMID- 8712732 TI - IL-2 gene therapy of advanced lung cancer patients. AB - We report here Phase I clinical-trial studies of retroviral-mediated interleukin 2 (IL-2) gene transfer to tumor-infiltrating lymphocytes that are re-infused to advanced lung cancer patients with pleural effusions. Ten lung cancer patients with malignant pleural effusions for whom all conventional therapy had failed were included in this Phase I protocol. Tumor infiltrating lymphocytes (TIL) from the patients were exposed to the retroviral plasmid pL(IL-2)SN containing the human IL-2 gene. Approximately 1-6 x 10(10) TIL cells transfected with IL-2 were re-infused into the chest cavity of each patient. The toxicity of this treatment with TIL/IL-2 gene therapy in these patients was minimal with transient slight fever of approximately 37.5. Pleural effusions did not re-accumulate for at least 4 weeks in six of ten patients. One patient was observed to have not only the resolution of the pleural effusions, but in addition the size of the original tumor decreased as seen by CT. The clinical results indicate this method of cancer gene therapy is safe and possibly efficacious against pleural effusions due to advanced lung cancer. PMID- 8712733 TI - Transforming growth factor alpha (TGF-alpha) expression in biopsies of colorectal carcinoma is a significant prognostic indicator. AB - BACKGROUND: Predicting the outcome of patients with colorectal adenocarcinoma (CRCA) prior to surgery would be valuable in selecting high-risk individuals who may benefit from pre-operative adjuvant therapy. The aim of this study is to determine whether the expression of TGF-alpha in preoperative biopsies of patients with CRCA constitutes a significant prognostic indicator. METHODS: We studied the expression of TGF-alpha in preoperative biopsies of 106 patients with CRCA, who had at least 5 years follow-up, using an anti-TGF-alpha monoclonal antibody and utilizing the ABC immunoperoxidase technique. For survival analysis, we used the actuarial survival method, and the Log Rank test for statistical significance. RESULTS: CRCAs with low TGF-alpha expression (less than 25% of the tumor cells immunoreactive for TGF-alpha) had a significantly poorer survival than those with high TGF-alpha expression (more than 25%). After excluding from analysis biopsies showing mucinous or poorly differentiated CRCA, known predictors of poor prognosis, the results remained significant (p = 0.0289). CONCLUSION: It is concluded, therefore, that low or absent expression of TGF alpha in pre-operative biopsies of patients with CRCA, as detected by immunohistochemistry, is a significant predictor of an unfavorable outcome. PMID- 8712734 TI - Effect of radiotherapy on serum level of interleukin 6 in patients with cervical carcinoma. AB - To determine the inflammatory state of cervical cancer after radiotherapy, we examined serum interleukin 6 levels (sIL-6) and C-reactive protein (CRP) before and after radiation therapy in sixteen patients with cervical cancer. External radiation therapy did not cause changes in sIL-6 and CRP during the examined periods. On the other hand, brachytherapy caused transient elevation of sIL-6 on the day after treatment by 6.39 +/- 1.89 pg/ml to 13.41 +/- 2.34 pg/ml (p < 0.05) while CRP did not show any significant change. Therefore, brachytherapy would induce a small inflammatory reaction. However, we confirmed that radiotherapy is a less invasive treatment than surgery from the point of view of cytokine related inflammation. PMID- 8712735 TI - Loss of heterozygosity and tumor suppressor gene mutations in chondrosarcomas. AB - Loss of heterozygosity (LOH) of chromosomes 13q and 17p and mutations of the retinoblastoma (Rb) and p53 gene were studied in 28 tumors from 22 patients with chondrosarcomas. Allele loss at polymorphic loci on 13q was found in 36% of tumors and the frequency was much higher in grade II (56%) or high grade (40%) tumors than in grade I tumors (18%). LOH of 17p, which was detected in 25% of all tumors, was of low frequency in low grade tumors (8% in grade I and 20% in grade II), whereas 80% of tumors with high grade tumors were positive for LOH. These observations may imply that LOH on 13q and 17p contributes to the tumor development at different stages. In low grade cases (grade I and II), tumor recurrence was observed more frequently in primary tumors with LOH on 13q (86%) than those without (8%), suggesting the significance of LOH analysis in the assessment of biological behavior of tumors. Structural alteration of the Rb gene was found in one dedifferentiated tumor, and point mutations of the p53 gene were found in all of five high grade tumors, indicating that high grade chondrosarcomas were genetically equivalent to other high grade sarcomas such as osteosarcomas. PMID- 8712736 TI - Conventional isolated hyperthermic antiblastic perfusion in the treatment of recurrent limb melanoma. AB - Hyperthermic Antiblastic Perfusion (HAP) is a widely used method for the treatment of recurrent limb melanoma. In terms of tumor response, locoregional control and survival HAP has led to better results than those achieved with any other treatment. The aim of this report is to analyze our own experience with HAP in locally advanced limb melanoma. Thirty-two patients were submitted to HAP. HAP lasted 60 minutes, with maximal local temperature of 40.5-42 degrees C using melphalan 10 mg/L limb volume as antiblastic agent. Twenty patients had in transit metastases and 12 local recurrence. Regional nodes were involved in 12 patients. Systemic leakage monitored with 125I or 99Tc ranged between 5-30% (mean 14%). No operative mortality nor major complications occurred. Local toxicity scored Wieberdink grade I in 8 patients, grade II in 17, grade III in 6 and grade IV in 1 case. Response rate (UICC) in the 16 patients treated with unexcised lesions was 94% (56% complete responses). With a median follow-up of 29 months (2 126) 14 patients relapsed after a median time of 10 months, and 17 patients are currently disease free, 3 of these are being reexcised and 3 re-perfused. Actuarial 5 years survival was 64%, with 39% disease free to the first relapse. Our results are consistent with the literature indicating HAP as a safe procedure with a high evidence of clinical responses. PMID- 8712737 TI - Prevention of radioinduced cystitis by orgotein: a randomized study. AB - On the basis of previous experiences indicating that the anti-oxidant agent Cu/Zn superoxide dismutase (SOD) is an effective drug in reducing acute and late radiation-induced tissue injury, in the Center of Radiotherapy and Oncology of Catalonia, Barcelona, Spain in 1990 we implemented a randomized prospective study to analyze the incidence and grade of side effects in a group of bladder cancer patients. After surgery patients were randomly allocated to receive either: Option A: Radiotherapy or Option B: Radiotherapy + SOD 8 mgr/IM/day, after each radiotherapeutic application. Between January 1990 and January 1995 a total of 448 patients were included (226 A/ 222 B). Apart from cutaneous side effects, a highly significant incidence of radioinduced acute cystitis and rectitis was detected in patients not treated by SOD. Which was similar to the delayed side effects. From our data we can conclude that SOD is effective in decreasing acute radioinduced damage, and also in preventing the appearance of more delayed disorders. PMID- 8712738 TI - Expression of the adhesion molecule CD44v3 is a prognostic factor in vulvar carcinoma. AB - Our aim was to evaluate the prognostic value of CD44v3 overexpression in vulvar cancer patients. Twenty-five cases of squamous cell carcinoma of the vulva with International Federation of Gynecology and Obstetrics (FIGO) stages I to III were examined for overexpression of the CD44 isoform CD44v3. The correlation of CD44v3 overexpression with clinical stage, histologic grade and overall survival was investigated. Follow-up ranged from 36 to 120 months. Clinical stage and histologic grade did not correlate with CD44v3 overexpression. CD44v3 overexpression was associated with poorer overall survival (log rank: P < 0.03). Immunohistochemically detected CD44v3 overexpression is significantly correlated with a reduced overall survival rate for patients with vulvar carcinoma. PMID- 8712739 TI - The pineal gland and cancer. AB - The pineal gland is considered today as one of the main organs involved in the transduction process which converts environmental light information into an endocrine response. The gland and its hormone melatonin seem to be important chronoimmunomodulators, and a reduction of the latter was associated with experimental and clinical immunodeficiencies and over the control of the neoplastic process. Moreover, melatonin can be an oncostatic or oncostimulating hormone, depending on the timing of its administration. The melatonin circadian rhythm is altered in cancer patients, and this rhythm could be modified as a consequence of certain therapies. Also Electromagnetic Fields (EMF) can affect the pineal function, perhaps working as synchronizers or, as this paper proposes, also through action of the "antenna effect" suggested for artificial human models, with a major energetic transfer over the cephalic area. The pineal could play an important role in the appearance and development of some forms of apparently EMF-related cancers. PMID- 8712740 TI - Urinary gonadotropin peptide in patients with cancer of digestive organs. AB - Urinary gonadotropin peptide (UGP) has been measured in gynecological and urological cancers, but its usefulness in the diagnosis of cancers of digestive organs has not been investigated. In this report, UGP was measured by sandwich enzyme immunoassay in 311 patients, including 166 patients with cancers of digestive organs and 43 healthy controls. Positive rates of UGP in various cancers of digestive organs were as follows: biliary tract 61.5%, pancreas 61.5%, esophagus 50.0%, liver 38.7%, colon and rectum 24.2%, and stomach 23.9%. The positive rate of UGP in benign diseases was 8.1%, and most false-positive patients were postmenopausal females. Positive rates of UGP were increased at advanced stages of gastric cancers, and UGP was decreased after tumor resection. From these results, it is suggested that UGP can be used as a tumor marker for the cancers of digestive organs. PMID- 8712741 TI - Analysis of p53 abnormalities in endoscopic gastric biopsies. AB - We performed immunohistochemical staining for p53 in 152 endoscopic gastric biopsy specimens, including 39 adenomas, 80 carcinomas, and 33 cases of regenerative atypia. Direct DNA sequencing of exons 5,7 and 8 of the p53 gene was performed on 8 specimens. Nuclear p53 immunoreactivity was observed in 36 of 80 carcinomas, 7 of 39 adenomas and none of the cases of regenerative atypia. Three of the adenomas demonstrated diffuse or focal p53 immunoreactivity. All three cases were associated with severe degrees of epithelial dysplasia, and two of them harbored carcinoma on subsequent polypectomy. Among 6 p53 positive carcinomas and adenomas sequenced, 3 showed DNA mutations, all G:C to A:T transitions. These results indicate that positive p53 immunoreactivity in intraepithelial atypical cells on gastric biopsies represents true dysplasia or carcinoma rather than regenerative atypia. PMID- 8712742 TI - Immunohistochemical evaluation of type IV collagenase (72-kd metalloproteinase) in prostatic intraepithelial neoplasia. AB - The aim of this study was to investigate the expression of type IV collagenase (72-kd metalloproteinase, MMP-2) in prostatic intraepithelial neoplasia (PIN) in relation to normal prostate (NP) and prostatic adenocarcinoma (PAc). Twenty formalin-fixed, paraffin-embedded prostatectomy specimens, in which NP, PIN and PAc were present, were immunohistochemically examined. The NP ducts and acini not contiguous with PIN and PAc showed slight MMP-2 immunostaining in the secretory cells, with some increase in intensity at the apical border, and moderate to strong immunoreactivity of some basal cells. In NP adjacent to PIN and PAc, rare ducts and acini showed strongly stained cells either isolated or in small groups of two, located within the thickness of the epithelium, close to the basement membrane. In the majority of PIN ducts and acini, the stratified secretory cells showed moderate staining. Most of these ducts and acini also showed strongly stained cells, which were mostly isolated, and either in contact with the basement membrane or scattered among the secretory cells. Low and high grade PIN showed some difference in the frequency of dark cells, which were more numerous in the latter. A small group of neoplastic acini adjacent to high grade PIN (early invasive adenocarcinoma) was observed in one of the 20 cases. Intense immunostaining was present in the acini originating from the PIN lesion. MMP-2 immunostaining of PAc was heterogeneous in intensity and location. Cribriform and solid/trabecular PAc showed weak cytoplasmic immunostaining; both moderately and intensely stained cells were seen in the cell layer adjacent to the stroma, intense immunostaining was shown by small clusters of neoplastic cells or single neoplastic cells located in the stroma. In acinar PAc, weak cytoplasmic immunostaining for MMP-2 was seen throughout most areas of the tumours, whereas moderately and intensely stained cells were observed less frequently than in cribriform and solid/trabecular adenocarcinoma. Intense immunostaining of single or small clusters of neoplastic cells located in the stroma was occasionally observed and, as with cribriform and solid/trabecular PAc, mainly located towards the periphery of the tumour nodules. Occasional ducts and acini with PIN and foci of PAc were either completely negative or very weakly stained. In conclusion, MMP 2 immunostaining increases progressively from NP, through PIN, up to invasive PAc. These results directly support the hypothesis that increased expression of metalloproteinases is a marker of malignant conversion. PMID- 8712743 TI - Ineffectiveness of sequential high dose methotrexate and 5-fluorouracil combined with epirubicin (FEMTX regimen) as a salvage therapy in advanced colorectal cancers and other gastrointestinal tumors. AB - The combination of sequential high dose methotrexate and 5-fluorouracil (FU) combined with epirubicin (FEMTX regimen) has shown some clinical efficiency as a first line therapy in advanced gastric cancer. We evaluated the therapeutic activity of this combination in advanced colorectal cancer patients who failed the FU + folinic acid regimen, and in advanced gastrointestinal cancer situation where no standard regimen is available. Twelve patients with measurable metastatic colorectal cancer refractory to first line (n = 6) or second line (n = 6) chemotherapy, as well as 9 patients with advanced gastrointestinal cancer, entered this pragmatic pilot study. Eighty-six per cent and 38% of the patients experienced severe haematological or non-haematological toxicities, respectively. Four patients were not evaluable for response due to their early departure from the scheme due to acute toxic events (n = 3) or toxic death (n = 1). No objective response was observed, but 9 stabilisations and 8 progressive diseases were noted. The FEMTX regimen is not of clinical value as a salvage therapy in colorectal and gastrointestinal cancer, because of its toxicity and its lack of efficacy. PMID- 8712744 TI - Relationship between thyroid-pituitary function and response to therapy in patients with recurrent breast cancer. AB - In this study, thyroid (T3, T4, free T3, free T4) and pituitary function (thyrotropin (TSH), growth hormone (GH), prolactin (PRL)) in 38 patients with recurrent breast cancer were examined. The patients were divided into three groups according to their response to the therapy. There were 16 partial response (PR), 10 no change (NC) and 11 progressive disease (PD) patients. The maximum and the minimum value for each hormone throughout the course of treatment were compared between three groups. The PD group showed significantly lower minimum T3 levels than the other two groups (p < 0.05). The maximum TSH level in the PD group was significantly higher than that of the other groups. The minimum TSH level in the PD group was significantly lower than that in the PR group (p < 0.05). The minimum TSH level in the NC group was also lower than that in the PR group. The maximum PRL level in the NC and the PD group was higher than that in the PR group (p < 0.05, p < 0.01, respectively). The tumors of the patients with temporal increase of TSH level were resistant to all subsequent therapies. These five patients died within four months followed by decreasing of the TSH level. It is concluded that thyroid and pituitary function, especially free T4, TSH and PRL, are predictive indicators of therapeutic response and the prognosis of the patients with recurrent breast cancer. PMID- 8712745 TI - Evaluation of two dose individualisation methods for carboplatin. AB - The pharmacokinetics of carboplatin are usually evaluated by measuring plasma concentrations of ultrafiltered platinum (UP). This approach, however may be less reliable than measuring the plasma concentration of total platinum (TP). In a group of 14 patients, which constituted a reference group, the clearance of TP was highly correlated with creatinine clearance, as estimated by the method of Cockroft and Gault. This relationship, together with only morphological and biological parameters, was used to estimate TP clearance, Vc and AUC, in a validation group of 8 patients. Estimated TP clearance was 97.9 +/- 18% of the actual value. The TP pharmacokinetic parameters of the reference group were used to estimate those of the validation group, using only two or three plasma concentration measurements (Bayesian approach). With the Bayesian approach, the estimated TP clearance was up to 99.9 +/- 2.7% of the actual value. In conclusion, estimation of TP pharmacokinetics may be reliably estimated as an alternative to UP in clinical practice. PMID- 8712746 TI - Multidrug resistance-associated protein (MRP) gene expression in human lung cancer. AB - Multidrug resistance-associated protein (MRP) is a 190 kD transmembrane protein and a potentially important drug-transporter protein in human cancers. While the MRP gene is expressed in normal cells and tissues, the expression in solid tumors is not sufficiently determined. MRP and mdr1 mRNA expressions were examined in normal lung parenchyma and in tumor tissues from six small cell lung cancer (SCLC) patients who had received preoperative chemotherapy and eleven nonsmall cell lung cancer (NSCLC) patients. The reverse transcriptase polymerase chain reaction was used. Normal lung tissues and all SCLCs expressed abundant levels of MRP mRNA, while the NSCLCs expressed a wide range of levels from low to high. Most tumor tissues coexpressed both MRP and mdr1, but the levels of mdrl expression was low except in two SCLCs and one NSCLC. MRP is more likely than mdr1 to be one of the clinical multidrug resistance mechanisms found in lung cancer. PMID- 8712748 TI - Electrophoretic pattern of serum glycoproteins on polyacrylamide disc gel in patients with breast cancer. AB - Numerous investigators have identified, isolated and characterized serum glycoproteins that are claimed to be specifically associated with malignancy. We have carried out serum glycoprotein electrophoresis on polyacrylamide disc gel in 53 breast cancer patients, at diagnosis as well as during and after therapy. Follow-up samples were divided into complete responders (CR) (n = 138) and nonresponders (NR) (n = 44). Glycoprotein electrophoresis showed multiple bands for each sample which were categorized into four groups: albumin, alpha, beta and gamma. The results revealed a decreasing number of CR and increasing number of NR with elevated (as compared to pretreatment levels) albumin fraction glycoproteins. Gamma region glycoproteins showed the reverse pattern to that of albumin region glycoproteins. The alpha and beta region glycoproteins revealed an increasing number of CR having higher values with increase in follow-up duration. In comparison with their pretreatment values CR showed significantly increased (Paired "t" test) values of albumin, alpha and beta region glycoproteins (p < 0.01, p < 0.001 and p < 0.001, respectively) and decreased gamma region glycoproteins (p < 0.001). The albumin, alpha, beta and gamma region glycoprotein levels were comparable between NR and untreated cancer patients. The variations in albumin, alpha, beta and gamma region glycoproteins correlate with treatment response, which might be useful in the treatment monitoring, and prediction of recurrence in breast cancer patients. PMID- 8712747 TI - Itraconazole can increase systemic exposure to busulfan in patients given bone marrow transplantation. GITMO (Gruppo Italiano Trapianto di Midollo Osseo). AB - Busulfan (BU) is an alkylating drug frequently used to prepare patients for bone marrow transplantation (BMT). Several studies have documented that there is important interpatient variability in BU disposition and systemic exposure, and that other drugs with a common metabolic pathway are capable of influencing BU clearance. We compared the BU pharmacokinetics and pharmacodynamics of 13 patients given BMT and receiving BU and itraconazole, with those of 26 matched controls who did not receive any anti-fungal agent, and with those of 13 matched patients treated with fluconazole as prophylaxis against fungal infections. The effect of itraconazole was best reflected in BU clearance since the BU dose was modified in some patients. BU clearance was decreased by an average of 20% in patients receiving itraconazole as compared to control patients and patients receiving fluconazole (p < 0.01). Mean BU clearance was 7.653 +/- 1.871 l/hr.m2 in the itraconazole patients, 10.103 +/- 2.007 l/hr.m2 in the fluconazole group and 9.373 +/- 1.702 l/hr.m2 in the control group. In this study itraconazole, but not fluconazole, markedly affected the pharmacokinetics of BU as an increase of BU plasma concentrations was observed. The nature of this interaction has not yet been fully characterized. Itraconazole and its analogues are inhibitors of both cytochrome P450 and lipoxygenase and since itraconazole can modulate BU pharmacokinetics, oxidative catabolism is probably a determinant of BU metabolism. This hypothesis should be tested in human metabolic studies. PMID- 8712749 TI - A prognostic score for prostatic adenocarcinoma based on clinical, histological, biochemical and cytometric data from the primary tumour. AB - The aim of this study was to create a multivariate prognostic score for prostatic adenocarcinoma. A retrospective analysis of clinical, histological and cytometric prognostic factors in 325 cases of prostatic adenocarcinoma followed up on average over 13 years was performed. A multivariate prognostic score was built by using the independent prognostic factors. M-category, T-category, Gleason score and patient age were independent prognostic factors in the entire cohort. In MO tumors T-category, density of tumor infiltrating lymphocytes, the presence of apoptotic cells, and patient age were independent prognostic factors. In T1-2MO tumors the density of tumour infiltrating lymphocytes, mitotic index, standard deviation of maximum nuclear diameter and the serum level of acidic phosphatase were independent prognostic factors. By combining the coefficients of the regression model, a prognostic score was built for each of the tumour categories. The subsequent survival analysis based on the prognostic score indicated that it is a highly significant prognostic factor superior to individual prognostic parameters. The results show that the combination of prognostic data is a valuable tool in assessing the correct prognostic category for prostatic adenocarcinoma. PMID- 8712750 TI - Evaluation of CYFRA 21-1 in malignant and benign pleural effusions. AB - CYFRA 21-1 was evaluated in 115 untreated patients with malignant pleural effusions (96 with primary lung cancer and 19 with non lung cancer) and 99 patients with benign pleural effusions. The levels of pleural fluid CYFRA 21-1 were from 1 to 385 times higher than those in serum, in all the examined patients. The mean level of pleural fluid CYFRA 21-1 was significantly higher in cancer patients than in patients with benign pleural effusion (96.1 ng/ml vs 26.2 ng/ml, p < 0.001). At 92% specificity for benign pleural effusion (> 50 ng/ml) the overall sensitivity of CYFRA 21-1 in malignant pleural effusions was 69.6%. When the histology was considered the highest sensitivity was found in squamous cell lung cancer (90%), followed by adenocarcinoma cell lung cancer (74%), non lung cancer (54%) and small cell lung cancer (25%). These results indicate that CYFRA 21-1 could be a useful pleural fluid marker in discriminating benign from malignant pleural effusion and particularly from those due to squamous and adenocarcinoma cell lung cancer. PMID- 8712751 TI - Antibody response to keyhole limpet hemocyanin (KLH) treatment in patients with superficial bladder carcinoma. AB - The dynamics of specific KLH-antibody production after intracutaneous and intravesical instillation was analysed. Nine patients (male, n = 7; female, n = 2, mean age 68.6 years, range 47-75) with primary superficial carcinomas of the bladder were intracutaneously immunized with 1 mg Keyhole limpet hemocyanin (KLH) after the complete resection of the tumors. Treatment was continued for 6 consecutive weeks, monthly for one year and thereafter bimonthly for 2 subsequent years, consisting of 20 mg KLH in 20 ml saline introduced intravesically. The antibodies against KLH in patient sera were determined by means of a specially developed direct enzyme-linked immunosorbent assay (ELISA; according to H. von der Kammer, Max Planck Institute for Biophysical Chemistry, Goettingen, Germany). Blood was taken for antibody-titer examination before treatment and 8 weeks after treatment. The KLH-antibody titer increased significantly (Mann-Whitney-Test P = 0.02) after KLH therapy in bladder cancer patients, however the level varied considerably from patient to patient. 6 of 9 patients (67%) presented increased serum antibody titers to KLH after immunotherapy. 4 patients (44.4%) remained free of tumor during the established follow-up period of 10-45 months (median 30.7 months). One patient without increased antibody titer to KLH was free of tumor, 2 patients however, suffered from tumor recurrence after the KLH course. 2 patients presented with tumor recurrence in spite of increased antibody titers. No evidence of tumor progression occurred in patients with recurrence after KLH therapy. 4 of 5 patients (80%) without tumor recurrence presented with a positive skin test. Of patients with tumor recurrence, 50% had a negative skin test. 44.4% KLH-treated patients had tumor recurrence The recurrence rate was 1.6. The time to recurrence was 8.75 months. KLH instillation did not induce major side effects. Positive skin test reactivity and KLH antibody response were more commonly seen in responding patients (i.e. those who remained tumor free after therapy) than in non-responders. The production of KLH antibodies, apparently is the biological response to the antigen stimulus of KLH. PMID- 8712752 TI - Hypermethylation of the myogenic gene Myf-3 in human breast carcinomas. AB - The methylation status of Myf-3 was studied in 34 human primary breast carcinomas and 9 normal breast tissues. One third of the carcinomas contained hypermethylated Myf-3. All normal tissues contained unmethylated Myf-3. Myf-3 hypermethylation was more common in poorly differentiated grade 3 tumours than in better differentiated grade 1 and 2 tumours. PMID- 8712753 TI - Tumor vascularization and prognosis in squamous cell carcinomas of the esophagus. AB - Quantification of tumor vascularization has recently been shown to be a parameter of potential prognostic significance in various types of malignant tumors. To determine the prognostic value of tumor vascularization in esophageal cancer, tumor samples from 150 patients with squamous cell carcinoma of the esophagus and 10 samples of normal esophageal mucosa were stained immunohistochemically with the monoclonal antibody QBEND/10 (CD34), which recognises endothelial cells. Using light microscopy, the number of microvessels was counted in the areas with the highest microvessel density (MVD). The microvessel density was significantly higher in the normal esophageal mucosa (mean: 130/mm2) than in the tumor samples (mean: 69/mm2, p = 0.0001). Correlation of the MVD in the tumor tissue with other prognostic factors showed significantly lower microvessel counts in tumors with lymphatic-vessel invasion (p = 0.0076) and in high pT-stages (p = 0.0081). No significant correlation was found between the MVD and pN stage, tumor size, tumor grade, blood-vessel invasion and proliferative activity. In the univariate survival analysis no significant differences were found between poorly vascularized tumors and highly vascularized tumors. A Cox proportional hazard regression selected the parameters lymphatic-vessel invasion (p = 0.0001), pT stage (p = 0.0034) and pN stage (p = 0.0256) but not MVD as independent prognostic variables. PMID- 8712754 TI - Increased serum 72 KDa metalloproteinase in serous ovarian tumors: comparison with CA 125. AB - 72 KDa metalloproteinase (MMP-2) is an enzyme present in neoplastic cells and also in normal fibroblasts. It specifically cleaves type IV collagen, and therefore may play a critical role in tumor invasion and metastasis mechanisms. The aim of the present study was to determine serum levels of MMP-2 in serous ovarian tumors, and compare these with serum levels of CA 125. Ten primary ovarian serous cystadenocarcinomas, 5 borderline tumors, and 10 serous cystadenomas, all treated with primary surgery, were recruited from our series of serous ovarian tumors, and studied. Patients' serum samples were obtained before surgery, and the MMP-2 levels were measured by the substrate capture enzyme linked immunosorbet assay. The analysis of serum MMP-2, gave values significantly higher in cystadenocarcinomas than in borderline tumors and cystadenomas (one way analysis of variance, P < 0.001); in particular, serum MMP-2 was significantly correlated to the MMP-2 immunostaining of the tumor (Spearman correlation, r = 0.82, and P < 0.001). An arbitrary cutoff of the median value of normal adult female samples (0.22 units) was chosen, and all except for one patient with cystadenocarcinoma was shown to have serum MMP-2 levels above the cutoff value, with 90% sensitivity, 70% specificity, and a 75% positive predictive value (50% of Cohen's Kappa); on the other hand, CA 125 showed 80% sensitivity, and a 73% positive predictive value. The association of serum MMP-2 with CA 125 increased sensitivity to 100% in patients with cystadenocarcinoma, with 70% persisting specificity and a 77% positive predictive value (54% of Cohen's Kappa). Serum MMP 2 levels were found to be significantly increased in patients with cystadenocarcinoma in comparison with borderline tumors and cystadenomas, showing a direct relationship with tissutal MMP-2 expression in serous ovarian tumors. Although our results were preliminary, they clearly suggested that serum MMP-2 may be an interesting diagnostic marker for cystadenocarcinomas. PMID- 8712755 TI - [Problems posed by asymptomatic pheochromocytoma and malignant tumors of the adrenal medulla]. AB - The authors analyse a group of 38 patients with a diagnosis of pheochromocytoma, in particular the topics of atypical symptomatology and malignancy. Three patients did not suffer from any subjective symptoms or other signs of pheochromocytoma ("incidentalomas"). Another three patients, although having giant tumours, only complained of atypical symptoms. Signs suspicious of malignancy were detected in 6 out of 38 patients. The authors recommend a practical approach to modern diagnostic and imaging tools for early primary diagnosis and for the early diagnosis of local recurrence and distant metastases. PMID- 8712756 TI - Pre-operative management for surgery of pheochromocytoma. AB - The authors analyze 15 patients who underwent pre-operative management prior to the pheochromocytoma removal. The purpose of this analysis is to evaluate the relation between clinical and haemodynamic criteria evaluating the level of alpha adrenergic blockade. The authors believe that pre-operative management based on clinical criteria does not necessarily ensure adequate alpha-adrenergic blockade. Invasive monitoring of haemodynamic variables is considered an important strategy for the quantification of actual haemodynamics status of patients with pheochromocytoma. PMID- 8712757 TI - Tumours of the adrenal cortex. AB - The authors report their experience of 34 benign and 17 malignant tumours of the adrenal cortex. Both occurred more frequently in women (79.4% of cortical adenomas and 70.6% of adrenocortical carcinomas). Moreover, females were significantly younger (average age: adenomas: females 44.9 years and males 54.4 years, adrenocortical carcinomas: females 34.1 years and males 58.3 years). Some tumours presented in the form of increased hormone production, while others were hormonally inactive and did not cause clinical signs until later. Pain was the first symptom in the cases of malignant adrenocortical tumours. Adenomas occurred as frequently twice in the left adrenal gland (24 versus 12), whereas carcinomas were more than twice are frequent on the right (10 versus 7). Ultrasonography and CT were appropriate and fully sufficient methods for the diagnosis of adrenal tumours. Arteriography was valuable in the differential diagnosis of large upper abdominal masses when the organ of origin could not be identified by CT and for determination of anatomic conditions and subsequent surgical tactics. For small tumours of the adrenal cortex, the classic lumbar approach through the bed of the resected eleventh rib is adequate. For larger tumours, extended lumbotomy, laparotomy or thoracotomy is necessary. PMID- 8712758 TI - [Malignant adrenocortical tumors. Apropos of 9 cases]. AB - The authors report 9 cases of adrenocortical carcinoma: 6 women and 3 men. 5 cases (55%) had a "nonfunctioning" tumor and 4 cases (45%) presented a clinical endocrinologic syndrome. According to the MacFarlane classification, patients were classified as: stage I: 1 case, stage III: 6 cases and stage IV: 2 patients. Adrenalectomy was performed in 7 cases. There was one postoperative death. 5 patients obtained clinical improvement. PMID- 8712759 TI - [Bilateral cysts of the adrenal glands. Apropos of a case]. PMID- 8712761 TI - [New diagnostic possibilities]. PMID- 8712760 TI - [A case of adrenal gland metastasis of a cutaneous melanoma treated by excision with 3-year survival]. AB - Case report of a 38-year-old man who, 14 months after resection of a malignant melanoma from the right shoulder in September 1990, presented with a tumour of the right adrenal gland. Resection confirmed metastatic melanoma. Chemotherapy was administered for 3 months. The patient was followed until his death in March 1995, i.e. three years after resection of the secondary adrenal tumour. PMID- 8712762 TI - [Detection of patients with pheochromocytoma using a cybernetic analysis method of the signs and symptoms of the disease]. AB - The diagnosis of pheochromocytoma is established more frequently in recent years due to better diagnostic possibilities. The aim of this study was to determine whether it is possible to distinguish the diagnosis of pheochromocytoma from mild, moderate and severe hypertension by analysis of symptoms and signs of pheochromocytoma using mathematical methods such as the pattern recognizing procedure before investigation of catecholamines in serum and urine. We retrospectively analysed the presence or absence of 45 symptoms and signs of pheochromocytoma except for investigation of catecholamines in 20 randomly selected patients with mild hypertension (HT), 20 patients with moderate HT, 20 patients with severe HT, and 20 patients with known pheochromocytoma. This procedure improved screening of pheochromocytoma and allowed early detection of the disease on a large scale. PMID- 8712763 TI - Role of phosphatidylinositol-linked proteins in paroxysmal nocturnal hemoglobinuria pathogenesis. AB - Patients with paroxysmal nocturnal hemoglobinuria have one or more mutant hematopoietic stem cell clones deficient in glycosylphosphatidylinositol (GPI) anchor synthesis owing to somatic mutations in the X-linked gene PIG-A. The progeny of mutant stem cells dominates the peripheral blood. The presence of a large number of GPI-anchor deficient, complement-sensitive erythrocytes leads to hemolytic anemia. The somatic mutations in PIG-A are small, various, and widely distributed in the coding regions and splice sites, indicating they occur randomly. Profiles of the mutations vary geographically, suggesting the presence of mutagen-induced mutations. The clonal dominance by the mutants does not seem to be solely due to the PIG-A mutation but may be caused either by autonomous expansion of the mutants due to a combination of the PIG-A mutation and some other genetic change(s) or by selection that preferentially suppresses normal stem cells. PMID- 8712764 TI - Ethical issues of fertility and reproduction. AB - How reproductive technologies are used has consequences not just for immediate users but for others as well. This article discusses issues that should be taken into consideration in coming to decisions about whether or not it is appropriate to use a particular reproductive technology, or whether to use a technology in a particular way. PMID- 8712765 TI - Gene transfer to hematopoietic stem cells: implications for gene therapy of human disease. AB - Transfer of new genetic material to hematopoietic stem cells and expression of the gene product in daughter cells of various lineages is an exciting approach to the treatment of congenital and acquired human diseases. This review summarizes the current status of retroviral and adeno-associated viral vectors for gene transfer to human hematopoietic cells, including extensive preclinical data as well as preliminary results from ongoing clinical trials. PMID- 8712766 TI - Pulmonary complications of HIV infection. AB - Pulmonary disease is a major source of morbidity and mortality in HIV-infected persons. Pneumocystis carinii pneumonia has decreased substantially during the last eight years, but in the United States it remains the most common disorder that announces the onset of AIDS. In contrast, tuberculosis is by far the most important AIDS-associated indicator disease in developing countries. Community acquired acute bacterial pneumonia is a common HIV-linked complication throughout the world; pneumonia occurs at all levels of immune suppression but increases in frequency as CD4 counts decrease. Fungal infections mainly afflict persons who live or have lived in the various endemic areas. AIDS-related Kaposi's sarcoma and lymphoma generally do not involve the lungs until the malignancies are advanced. The increasing use of successful chemoprophylaxis against many important HIV-associated infections is increasing the incidence of other end stage complications such as cytomegalovirus and disseminated MAC disease. PMID- 8712767 TI - Complications of laxative abuse. AB - Laxative abuse is an uncommon but clinically important cause of chronic diarrhea, a condition often associated with other gastrointestinal symptoms, as well as with disturbances in electrolyte and acid-base balance. A high index of suspicion, a detailed history, and the detection of laxative in stool and/or urine will establish the diagnosis once routine laboratory, endoscopic, and radiologic investigations have excluded common causes of chronic diarrhea. Confirmation of the diagnosis may rule out the need for more extensive, invasive, and costly investigations. However, management is frequently difficult owing to the laxative abuser's complex underlying psychopathology. PMID- 8712768 TI - The cardiac ion channels: relevance to management of arrhythmias. AB - The electrical activity of cardiac tissue is determined by the highly regulated flow of ions across the cell membrane during the cardiac action potential. Ion channels are pore-forming proteins through which these electric currents flow. In this review, the ion currents that underlie the action potential are first described. Then, the way in which expression of individual ion-channel genes results in such ion currents is discussed. Finally, the concept that arrhythmias may be due to abnormalities of structure, function, or number of ion channels, or the way in which they respond to abnormalities in their environment (such as acute ischemia), is reviewed. Further understanding of the molecular mechanisms underlying normal and abnormal cardiac electrophysiologic behavior should allow the development of safer and more effective antiarrhythmic interventions. PMID- 8712769 TI - Alcohol, coronary disease, and hypertension. AB - Disparities in the relationships between alcohol consumption and various cardiovascular conditions are now evident, with complex interrelationships between conditions. An inverse relationship of alcohol use to coronary heart disease is supported by many population studies. Interpretation of these data as a protective effect of alcohol against coronary disease is strengthened by plausible mechanisms. Although some experimental data suggest the hypothesis that wine, in particular, has additional protective benefit, prospective studies show no consensus on this point. Strong, consistent epidemiologic data support a relationship of heavier drinking to hypertension. Intervention studies show a pressor effect of alcohol, which appears and regresses within several days, but a mechanism has not yet been established. As with most aspects of alcohol and health effects, the data do not suggest monotonic relationships of alcohol with these conditions. Thus, amount of alcohol taken is a crucial consideration. Advice to concerned persons needs to take into account individual factors in drinkers or potential drinkers. PMID- 8712770 TI - Fatal asthma. AB - This brief review highlights the rise in asthma mortality witnessed in many countries. It outlines the multifactorial character of this phenomenon, which in most instances is a complex interplay between physician, patient, and environment. The pathophysiology of airflow obstruction and in particular the role of hyperinflation are outlined. These data may allow better prediction of who is at risk of fatal asthma (FA). Finally, potential approaches to the prevention of FA are outlined. PMID- 8712771 TI - Antimicrobial-resistant pathogens in the 1990s. AB - Streptococcus pneumoniae, Enterococcus faecalis, Enterococcus faecium, Staphylococcus aureus, and Klebsiella pneumoniae have become increasingly resistant to antimicrobial agents. This chapter reviews the epidemiology of this resistance, its detection in the laboratory, the mechanisms of resistance, and the options for therapy and infection control. PMID- 8712772 TI - Sun exposure and skin disease. AB - Sunlight exposure produces a variety of adverse cutaneous effects. Erythema, photosensitivity, and immunologic alterations represent acute events, whereas photoaging and carcinogenesis are long-term consequences. These adverse cutaneous sequelae can be minimized by photoprotection in the form of sun avoidance, regular cover-up with clothing, and sunscreen application. This chapter reviews the diagnosis and treatment of sun-related skin disorders and recommendations for reducing photodamage. PMID- 8712773 TI - Harnessing the power of the placebo effect and renaming it "remembered wellness". AB - The placebo effect yields beneficial clinical results in 60-90% of diseases that include angina pectoris, bronchial asthma, herpes simplex, and duodenal ulcer. Three components bring forth the placebo effect: (a) positive beliefs and expectations on the part of the patient; (b) positive beliefs and expectations on the part of the physician or health care professional; and (c) a good relationship between the two parties. Because of the heavily negative connotations of the very words "placebo effect, " the term should be replaced by "remembered wellness." Remembered wellness has been one of medicine's most potent assets and it should not be belittled or ridiculed. Unlike most other treatments, it is safe and inexpensive and has withstood the test of time. PMID- 8712774 TI - Trinucleotide instability: a repeating theme in human inherited disorders. AB - In recent years, a completely new mechanism of mutation has emerged in a number of disorders that display perplexing and paradoxical features of genetic inheritance. This mechanism involves the expansion and intergenerational instability of stretches of consecutive identical nucleotide triplets that also exist as shorter stable segments on normal chromosomes. The unstable nature of the trinucleotide segments has solved many of the genealogic puzzles in these disorders and has provided a new tool for predictive testing. Treatments for the disorders await a better understanding of the different pathogenic processes that are triggered by various expanded repeats. The existence of numerous other disorders with peculiarities of genetic inheritance suggests that this mutational mechanism may be a major cause of human inherited disease. PMID- 8712775 TI - Myocardial preconditioning promises to be a novel approach to the treatment of ischemic heart disease. AB - In the phenomenon termed "ischemic preconditioning," a brief period of ischemia prior to a more prolonged one improves myocardial function (after reperfusion) and diminishes infarction. This phenomenon has been described extensively in experimental animals and now in humans. It is triggered by several agents released by ischemic cells and can be reproduced by infusion of agonists coupled to protein kinase C (PKC), e.g. adenosine, angiotensin, phenylephrine, bradykinin, and endothelin. The intracellular signaling pathway involves a phospholipase, either C or D, which metabolizes membrane phospholipids to produce diacylglycerol, a necessary endogenous cofactor for PKC activation. Which protein(s) is phosphorylated by PKC is not yet known, nor is the identity of the end-effector that actually mediates protection of the ischemic cell. Identification of the end-effector may make it possible in the routine treatment of patients with ischemic heart disease to precondition and thereby salvage ischemic myocardium and improve survival. PMID- 8712776 TI - Management of patients with intermediate-thickness melanoma. AB - It is important to understand the management of patients with melanoma because of a rising incidence of this cancer in the United States. The most important prognostic factor is tumor thickness. Patients with early melanoma (thickness < 1.0 mm) have an excellent prognosis and are effectively treated with narrow local excision (1-cm radius). Those with advanced melanomas (tumor thickness > 4.0 mm) have a high risk of metastases and are treated with a wider local excision (2-3 cm). Controversies surrounding the management of patients with intermediate thickness melanoma (thickness 1-4 mm) center on the issues of local excision and management of regional lymph nodes. Randomized trials have shown that a 2-cm radius of excision will minimize the risk for local recurrence. Although retrospective analyses have indicated a survival advantage for elective regional lymph node dissection, two randomized trials have not shown a benefit for the early removal of lymph nodes. Two more randomized trials are in progress. Techniques are now available to identify the sentinel lymph node, which will help to limit lymph node dissections to those patients with nodal metastases. Postoperative surveillance for recurrence is most effectively performed with a combination of patient self-examination and regular physician examinations rather than frequent X-rays and laboratory tests. Adjuvant therapy has not been proven effective for melanoma patients; however, many investigational trials are available. PMID- 8712777 TI - Nitric oxide synthase: role as a transmitter/mediator in the brain and endocrine system. AB - Nitric oxide is a unique biological messenger molecule. It is produced by endothelial cells to mediate blood vessel relaxation; it mediates, in part, the immune functions of activated macrophages; and in the central and peripheral nervous system it serves as a neurotransmitter. In the nervous system, nitric oxide may regulate neurotransmitter release, it may play a key role in synaptic plasticity and morphogenesis, and it may regulate sexual and aggressive behavior. Under conditions of excessive formation, nitric oxide is emerging as an important neurotoxin. PMID- 8712778 TI - The molecular basis of X-linked severe combined immunodeficiency: defective cytokine receptor signaling. AB - X-linked severe combined immunodeficiency (XSCID) is an inherited disease characterized by profoundly diminished cell-mediated and humoral immunity. XSCID was found to result from mutations in the interleukin-2 (IL-2) receptor gamma chain. Knowledge of the genetic defect has important implications for prenatal and postnatal diagnosis, carrier female identification, and the possibility of gene therapy. The fact that the phenotype and clinical manifestations in XSCID are more severe than the abnormalities found in humans or mice deficient in IL-2 led to the speculation and subsequent confirmation that the IL-2 receptor is not the only receptor to contain the gamma chain. Instead, the gamma chain is also a component of the receptors for IL-4, IL-7, IL-9, and IL-15 and is now denoted as the common cytokine receptor gamma chain, gamma c. The role of gamma c in signaling and lymphoid development and the implications of a shared receptor component are discussed. PMID- 8712779 TI - Explaining fatigue in congestive heart failure. AB - Fatigue is a prominent symptom in patients with chronic heart failure, limiting physical activity and impairing quality of life. Although the underlying mechanisms are not clearly identified, alterations associated with peripheral adaptation in heart failure appear to play an important role, including a variably impaired peripheral perfusion during exercise, reduced oxidative capacity of skeletal muscle, impaired muscle strength, and possibly reflex mechanisms associated with alterations in the metabolism of skeletal muscle. Exercise training can, in part, reverse these peripheral alterations, improve exercise capacity, and alleviate fatigue. PMID- 8712780 TI - Hepatitis E: an overview. AB - Hepatitis E has a worldwide distribution and causes substantial morbidity and mortality in some developing countries, particularly among pregnant women. Hepatitis E virus (HEV) has recently been cloned and sequenced and new diagnostic tests have been developed; these tests have been used to begin to characterize the natural history and epidemiologic features of HEV infection. Experimental vaccines have also been developed that offer the potential to prevent hepatitis E. However, to develop effective strategies to prevent this disease, much remains to be learned about HEV, including the vehicles of transmission, the reservoir(s) of the virus, and the natural history of protective immunity. PMID- 8712781 TI - High-altitude pulmonary edema: current concepts. AB - High-altitude pulmonary edema (HAPE) occurs in unacclimatized individuals who are rapidly exposed to altitudes in excess of 2450 m. It is commonly seen in climbers and skiers who ascend to high altitude without previous acclimatization. Initial symptoms of dyspnea, cough, weakness, and chest tightness appear, usually within 1-3 days after arrival. Common physical signs are tachypnea, tachycardia, rales, and cyanosis. Descent to a lower altitude, nifedipine, and oxygen administration result in rapid clinical improvement. Physiologic studies during the acute stage have revealed a normal pulmonary artery wedge pressure, marked elevation of pulmonary artery pressure, severe arterial unsaturation, and usually a low cardiac output. Pulmonary arteriolar (precapillary) resistance is elevated. A working hypothesis of the etiology of HAPE suggests that hypoxic pulmonary vasoconstriction is extensive but not uniform. The result is overperfusion of the remaining patent vessels with transmission of the high pulmonary artery pressure to capillaries. Dilatation of the capillaries and high flow results in capillary injury, with leakage of protein and red cells into the alveoli and airways. HAPE represents one of the few varieties of pulmonary edema where left ventricular filling pressure is normal. PMID- 8712782 TI - Clinical implications of the p53 gene. AB - The capacity for malignant growth is acquired by the stepwise accumulation of defects in specific genes regulating cell growth and tissue homeostasis. Although several hundred genes are known to control growth, molecular genetic studies in cancer show that few of these are consistently involved in the natural history of human cancer, and those typically in only certain types of malignancy. Prospects for development of molecular-based diagnostic and therapeutic strategies with widespread application did not look promising, until it was realized that the p53 tumor suppressor gene was defective in approximately half of all malignancies. This discovery generated research efforts of unparalleled intensity to determine how p53 functions at the molecular level, and how to apply this knowledge to clinical ends. PMID- 8712783 TI - Antineutrophil cytoplasmic antibodies in the classification of vasculitis. AB - Two important types of antineutrophil cytoplasmic antibodies (ANCA) have been identified: anti-proteinase 3 and anti-myeloperoxidase antibodies. In the appropriate clinical setting, the presence of either is virtually diagnostic of the subset of vasculitis that includes Wegener's granulomatosis, microscopic polyangiitis (microscopic polyarteritis), the Churg-Strauss syndrome, idiopathic pauci-immune necrotizing and crescentic glomerulonephritis, and related and overlapping forms of these vasculitidies. The finding of ANCA throughout this group identifies these syndromes as belonging to a single category or spectrum of disease. PMID- 8712784 TI - Complications of laparoscopic cholecystectomy. AB - Video technology allows surgeons to offer patients operations through small incisions. While these minimally invasive techniques have been employed for a wide range of operations for many years, only recently has a laparoscopic cholecystectomy been possible. Compared to the same operation performed via laparotomy, it is deemed the "gold standard." Overall mortality after laparoscopic cholecystectomy ranges from 0-1%, and the rate of major complications is less than 5%. As a group, general surgeons are beyond their initial learning curve for laparoscopic cholecystectomy, but the majority of iatrogenic injuries can be successfully avoided by appreciating the limitations and pitfalls of laparoscopic surgery, and by carefully dissecting the hepatocystic triangle before dividing any structure. Early identification and management of complications will minimize potentially devastating complications of small, unrecognized injuries. Physicians need to accurately advise patients of the proposed advantages and potential problems of laparoscopic cholecystectomy before referring them to a surgeon. PMID- 8712785 TI - Role of endothelium in thrombosis and hemostasis. AB - Vascular endothelium is strategically located at the interface between tissue and blood. It is pivotal for protecting against vascular injury and maintaining blood fluidity. Normal endothelium releases prostacyclin and nitric oxide, potent inhibitors of platelet and monocyte activation and vasodilators. Their syntheses are governed by isoforms of enzymes. Normal endothelial surface expresses ecto adenosine diphosphatase, which degrades adenosine diphosphate and inhibits platelet aggregation; thrombomodulin, which serves as a binding site for thrombin to activate protein C; and heparin-like molecules, which serve as a cofactor for antithrombin III. Normal endothelium secretes tissue plasminogen activator, which activates the fibrinolysis system. Endothelium produces and secretes von Willebrand factor, which mediates platelet adhesion and shear-stress-induced aggregation. Injury to endothelium is accompanied by loss of protective molecules and expression of adhesive molecules, procoagulant activities, and mitogenic factors, leading to development of thrombosis, smooth muscle cell migration, and proliferation and atherosclerosis. PMID- 8712786 TI - The geneticist's approach to complex disease. AB - Many studies are in progress worldwide to elucidate the genetics of complex diseases. Nevertheless, few articles are available that provide the scientific rationale and give guidelines for such ambitious endeavours. We describe the methodology and background necessary to study the genetics of complex disease and discuss how to analyze the data. We also provide a table of some ongoing studies. In particular, we wish to emphasize the analysis of intermediate, heritable, quantitative traits as a means of dissecting the genetic basis of a complex trait. PMID- 8712787 TI - Bartonella (Rochalimaea) infections: beyond cat scratch. AB - Five species of Bartonella have been found to infect humans, henselae, quintana, elizabethae, bacilliformis, and vinsonii. The most common of these in North America are Bartonella quintana and Bartonella henselae, the agents of trench fever, bacillary angiomatosis, and parenchymal peliosis, and in the case of B. henselae cat-scratch disease. B. bacilliformis is endemic in Peru and Ecuador, where it causes oroya fever or Carrion's disease. New methods of diagnosing Bartonella infections have resulted in increased recognition of the many manifestations of these infections. Early recognition is crucial, as these are potentially fatal opportunistic infections that usually respond rapidly to appropriate antimicrobial therapy. PMID- 8712788 TI - The vascular biology of nitric oxide and its role in atherogenesis. AB - Nitric oxide (NO), the biologically active component of endothelium-derived relaxing factor, has critical roles in the maintenance of vascular homeostasis. Decreased endothelial NO production, as a result of endothelial dysfunction, occurs in the early phases of atherosclerosis. NO appears to inhibit atherogenesis by inhibiting leukocyte and platelet activation and by inhibiting smooth muscle cell proliferation. Endothelial denudation is a prominent feature of vascular injury associated with percutaneous angioplasty, and decreased NO production appears to contribute to the restenosis process. Manipulation of the NO/cGMP signal transduction system may provide novel therapeutic approaches for limiting atherogenesis and neointimal proliferation in the future. PMID- 8712789 TI - Goodpasture's disease and Alport's syndromes. AB - Goodpasture's, or anti-glomerular basement membrane, disease is an uncommon, usually severe disease caused by autoimmunity to a component of certain basement membranes. Alport's syndrome is an inherited, degenerative disorder that affects specific basement membranes. The two are linked by the involvement of type-IV collagen (basement membrane collagen) in their pathogenesis. PMID- 8712790 TI - Apolipoprotein E alleles as risk factors in Alzheimer's disease. AB - Alzheimer's disease (AD) is unique in medicine in that millions of people suffer from what appears to be the same form of disease, and unlike most other late onset diseases, the genetic etiologies have been well identified. Three early onset forms of AD inherited as autosomal dominant traits account for less than 2% of prevalent AD. A major susceptibility locus, apolipoprotein E (APOE, gene; apoE, protein) is associated with risk and age of onset distributions for the common familial and sporadic late-onset AD. The identification of additional genetic susceptibility genes in the etiology of AD and the metabolic mechanisms leading to differences in age of onset and disease pathogenesis are active areas of current research. PMID- 8712791 TI - The role of anti-inflammatory drugs in the prevention and treatment of Alzheimer's disease. AB - Risk factor intervention is a useful strategy for prevention of poorly understood diseases. Fifteen studies have examined the relation of glucocorticoid and nonsteroid antiinflammatory treatments and onset or progression of Alzheimer's disease (AD). Fourteen of these studies suggest that such treatments (especially nonsteroidal agents) prevent or ameliorate symptoms of AD. Abundant circumstantial evidence implicates inflammation in the pathogenesis of AD. Inhibition of cyclooxygenases, the central action of nonsteroidal antiinflammatory drugs (but not a prominent effect of steroids), limits inflammation, but it may also alter neural metabolic pathways, resulting in cell death from excitotoxicity or oxidative stress. Randomized controlled trials are needed to determine whether steroids, nonsteroidal antiinflammatory drugs, or both can prevent or treat the symptoms of AD. PMID- 8712792 TI - Endothelial-dependent mechanisms in chronic inflammatory leukocyte recruitment. AB - Peripheral blood leukocytes interact with the vascular endothelium in a wide range of physiologic and pathophysiologic situations. A current working concept is that activation of vascular endothelium is an important event during the inflammatory response, conferring spatial and temporal localization and leukocyte type selectivity to the recruitment process. This chapter highlights recent advances in our understanding of the endothelial-dependent molecular mechanisms that mediate recruitment of mononuclear leukocytes (lymphocytes and monocytes) and discusses these advances in the context of chronic inflammatory diseases and their potential therapeutic interventions. PMID- 8712793 TI - The role of lipoprotein(a) in atherogenesis and thrombosis. AB - Lipoprotein(a) [Lp(a)] represents an important independent risk factor for atherosclerotic cardiovascular disease. Lp(a) constitutes a class of low-density lipoprotein-like particles that are structurally heterogeneous due to variability within the distinguishing apoprotein, apolipoprotein(a) [Apo(a)]. Apo(a) bears a high degree of homology to the fibrinolytic zymogen, plasminogen, the parent molecule of the serine protease plasmin. Apo(a) contains a variable number of tandemly repeated triple-loop units called kringles, which appear to mediate Lp(a)'s interactions with fibrin and cell surface receptors. Although the mechanism of its atherogenicity is unknown, Lp(a) has been implicated in the delivery of cholesterol to the injured blood vessel, in blockade of plasmin generation on fibrin and cell surfaces, and as a stimulus for smooth muscle cell proliferation. In addition, new members of the plasminogen/Apo(a) gene family have been defined, creating a potential link between Lp(a) and the control of angiogenesis in both health and disease. Pharmacologic therapy of elevated Lp(a) levels has been only modestly successful; apheresis remains the most effective therapeutic modality. PMID- 8712794 TI - Differential diagnosis and management of Cushing's syndrome. AB - The diagnosis of endogenous Cushing's syndrome requires demonstration of an increased cortisol secretion rate, best achieved by urinary free cortisol excretion determinations. In borderline or confusing cases, loss of diurnal cortisol rhythmicity, a combined dexamethasone/corticotropin releasing hormone (CRH) test, or close monitoring of the patient for a few months will be helpful in ruling out pseudo-Cushing's. Primary adrenal Cushing's syndrome can be ruled out on the basis of a normal or elevated basal and/or CRH-stimulated plasma adrenocorticotropin (ACTH) and a negative adrenal computed tomography. ACTH dependent Cushing's syndrome can then be differentiated on the basis of a CRH test and imaging procedures. A discrete pituitary lesion on magnetic resonance imaging and a standard CRH test with results consistent with such a lesion are sufficient to proceed to transsphenoidal surgery. If no discrete pituitary lesion is present, or if the CRH test is equivocal, bilateral simultaneous inferior petrosal sinus sampling with CRH administration is necessary to distinguish between a pituitary and an ectopic source. Surgery is the treatment of choice for all types of Cushing's syndrome. In the few cases in which transsphenoidal surgery fails or the disease recurs, repeat transsphenoidal surgery, or radiation therapy in association with mitotane treatment, is a reasonable alternative. Bilateral adrenalectomy effectively cures hypercortisolism if resection of the ACTH-secreting tumor is unsuccessful and radiation/medical therapy fails. PMID- 8712795 TI - The evaluation of diagnostic tests: principles, problems, and new developments. AB - This article describes the use of probability in reasoning about diagnostic test results and the importance of accurate measures of test performance. Most studies of test performance are retrospective analyses of unplanned observations. Retrospective studies are likely to result in biased estimates of test performance. This article shows how to adjust for biases in the results of retrospective studies and how to design a prospective study that will accurately measure the sensitivity and specificity of a test. PMID- 8712796 TI - How does lithium work on manic depression? Clinical and psychological correlates of the inositol theory. AB - How lithium works in manic-depressive illness is unknown. Recently, however, a powerful hypothesis has been gaining momentum. Distinguished by its testability and clinical implications, the inositol depletion hypothesis of lithium action is relevant to treatment of lithium side effects, to the development of new compounds with the clinical profile of lithium, and to new experimental treatment of depression. PMID- 8712797 TI - The epidemiology of psychiatric disorders and the de facto mental health care system. AB - Recent national epidemiologic studies have provided data on the number of people in the United States with mental and addictive disorders. Many of these people receive their care in the general medical care sector. This has important implications for diagnosis and treatment of mental and addictive disorders. PMID- 8712798 TI - The immunotherapy of solid cancers based on cloning the genes encoding tumor rejection antigens. AB - Cellular immune reactions play a major role in the host reaction to growing cancers. Tumor infiltrating lymphocytes (TIL) can be isolated from melanomas and can specifically recognize unique tumor antigens. The adoptive transfer of TIL plus interleukin-2 can mediate tumor regression in patients with metastatic melanoma. TIL capable of mediating tumor regression have been used to clone and sequence a variety of the genes that encode the tumor-regression antigens recognized by these TIL. This information has opened new opportunities for the development of cancer immunotherapies. These gene products can be used to generate lymphocytes, in vitro, with improved antitumor activity for use in adoptive transfer. Active immunization can be performed using either the immunodominant peptides present in these proteins or by incorporating the tumor antigen genes into recombinant viruses. Cancer vaccine trials using many of these approaches have recently begun. Attempts to apply a similar strategy to epithelial tumors such as breast and ovarian cancer are underway. PMID- 8712799 TI - Nicotine addiction and treatment. AB - The persistence of cigarette smoking despite widespread awareness of adverse health effects results from an underlying addiction to nicotine. Unaided attempts to quit smoking are generally unsuccessful. This article discusses nicotine addition and therapeutic techniques that have been or are being developed to relieve smoking withdrawal symptoms and promote abstinence from smoking. These techniques include nicotine chewing gum, skin patches, nasal sprays, and inhalers, as well as pharmacotherapies such as mecamylamine and clonidine, serotonergic treatments such as buspirone, and antidepressants such as buproprion. A nondrug approach using cigarette substitutes that mimic the airway sensations produced by cigarette smoke is also discussed. PMID- 8712800 TI - Genetics of non-insulin-dependent (type-II) diabetes mellitus. AB - Both genetic and environmental factors contribute to the etiology of non-insulin dependent diabetes. The genetic component is heterogeneous and in some patients is probably complex, involving multiple genes. Specific genetic defects have been identified for rate monogenic forms of NIDDM: maturity-onset diabetes of the young, or MODY (which is due to glucokinase mutations in about 40% of families), syndromes of extreme insulin resistance (which often involve the insulin receptor), and diabetes-deafness syndromes (with defects in mitochondrial genes). In contrast, the genes involved in common forms of NIDDM are still uncertain. Mutations have been extensively searched in genes regulating insulin signaling and secretion. Some evidence of involvement has been produced for insulin receptor substrate-1, glycogen synthase, the glucagon receptor, a ras-related protein (Rad), histocompatibility antigens, PC-1, and fatty acid binding protein, but the contributions of these genes to NIDDM is probably small. Other candidate genes (e.g. insulin, insulin receptor, glucose transporters) have been excluded as major diabetogenes. New insights are expected in the near future from the systematic scanning of the genome for linkage with NIDDM. PMID- 8712801 TI - The lupus anticoagulant/antiphospholipid syndrome. AB - Lupus anticoagulants and anticardiolipin antibodies have been strongly associated with the risk of thrombosis, recurrent fetal loss, thrombocytopenia, and a number of other clinical manifestations that together have been referred to as the antiphospholipid syndrome. Despite growing evidence of the significance of this relationship, the pathogenetic mechanisms involved are largely unknown. Recent data suggest strongly that lupus anticoagulants (LACs) and anticardiolipin antibodies (ACAs) are antibodies to protein-phospholipid complexes rather than to phospholipids, as had originally been thought, and that other protein phospholipid complexes, not recognized by standard assays for LACs or ACAs, may also exist in patients with the antiphospholipid syndrome. Although very recent experimental data may lead to new therapeutic approaches in this syndrome, at present we can only deal with the thrombotic risk by the use of long-term anticoagulation. This chapter reviews current methods of diagnosis, concepts of pathogenesis, and the basis for an approach to anticoagulation in patients at risk for thrombosis or other manifestations of the antiphospholipid syndrome. PMID- 8712802 TI - Pathogenesis and management of acute promyelocytic leukemia. AB - Acute promyelocytic leukemia, formerly a highly lethal disease, was recently shown to be highly responsive to treatment with the natural retinoid, all-trans retinoic acid. This compound induces terminal differentiation of the malignant cells and produces complete remission in most patients with this disease. The clinical response is due in part to the presence of a chromosomal break that occurs in a gene that encodes a nuclear retinoic acid receptor. PMID- 8712803 TI - Endocrine tumors of the gastrointestinal tract. AB - Endocrine tumors of the gastrointestinal tract produce a variety of secretory products that cause unique clinical syndromes. Diagnosis, which is often delayed, requires a strong index of suspicion and must be confirmed by biochemical tests. Precisely where these tumors originate remains a topic of controversy. However, several growth factors that may be involved in tumor development have been identified, and genetic abnormalities in patients with multiple endocrine neoplasia have been described. New pre- and intraoperative localization techniques have greatly increased the ability to identify and resect these tumors. The long-acting somatostatin analogue octreotide is frequently useful as a tracer to localize tumors and as symptomatic therapy for limiting release of secretory products produced by the tumors. In some instances it may also have direct anti-tumor activity. PMID- 8712804 TI - An integrated view of beta-cell dysfunction in type-II diabetes. AB - Type-II (non-insulin-dependent) diabetes mellitus (NIDDM) is a heterogeneous disease resulting from insulin resistance and beta-cell dysfunction. beta-Cell dysfunction in Type-II diabetes is characterized by a specific lack of first phase glucose-induced insulin secretion. This defect is readily reversible upon normalization of blood glucose levels. Chronic hyperglycemia itself is harmful to the beta-cell and affects both insulin biosynthesis and exocytosis. No unique intracellular defect has been demonstrated to be responsible for all common forms of the disease. However, mutations of the glucokinase gene have been identified in maturity onset diabetes in the young, a particular form of NIDDM. PMID- 8712805 TI - The current status of the implantable cardioverter defibrillator. AB - Since its initial application in 1980, the implantable cardioverter defibrillator (ICD) has evolved into a highly sophisticated device that offers programmable tiered therapy (antitachycardia pacing, cardioversion, and defibrillation) for ventricular tachyarrhythmias and backup pacing for bradycardia. It also provides diagnosis information about detected events. Successful implantation of nonthoracotomy systems is now routine. The availability of nonthoracotomy systems has reduced perioperative morbidity and mortality as well as hospital costs. Although the ICD has virtually eliminated sudden cardiac death in recipients, overall cardiac mortality remains considerable in these patients. Several prospective randomized clinical trials comparing ICD therapy to antiarrhythmic drug therapy are under way. PMID- 8712806 TI - Diagnosis and management of nonfunctioning pituitary tumors. AB - It is now recognized that most clinically nonfunctioning pituitary tumors express gonadotropin hormones or their subunits in vitro, and sometimes in vivo. Many of these tumors are diagnosed by sensitive imaging techniques while patients are still asymptomatic. We outline an overview of the diagnosis and clinical management of these tumors. PMID- 8712807 TI - Clinical issues. PMID- 8712808 TI - Surgical management of patients with severe head injuries. AB - Minutes can make the difference between life and death when patients with severe head injuries require surgery. Subdural, epidural, and intracerebral hematomas and cerebral contusions and gunshot wounds are the pathologic entities encountered most frequently during emergency surgery in patients with severe head injuries. Neurosurgical team members frequently use hyperventilation, mannitol and barbiturates, and sophisticated monitoring modalities to manage patients with severe head injuries during and after surgery. Although monitoring a patient's intracranial pressure (ICP) through a ventriculostomy catheter remains the most widely used gauge of cerebral metabolism, neurosurgical teams also are using fiber-optic ICP monitoring catheters, cerebral blood flow measurement probes, microdialysis catheters, jugular venous oxygen saturation catheters, and brain oxygen content measurement electrodes. Coordinated teamwork by perioperative nurses, neurosurgeons, anesthesia care providers, and emergency department staff members helps ensure the best possible outcomes for patients who require surgery for management of severe head injuries. PMID- 8712809 TI - Surgical treatment of patients with open tibial fractures. AB - Open tibial fractures are true surgical emergencies because of the risk of extensive infection to bone and devitalized soft tissue. The most serious consequence of open tibial fractures is osteomyelitis, which usually can be prevented by prompt surgical intervention within six to eight hours after injuries occur. Open tibial fractures often are the result of trauma from motor vehicle collisions, farm accidents, falls from heights, or gunshot wounds. Initial management of patients with multiple trauma injuries focuses on their life-threatening injuries before or during orthopedic surgical intervention for open tibial fractures. Orthopedic surgeons often work in collaboration with general, vascular, and plastic surgeons and perform multiple surgical procedures (eg, fasciotomy procedures for compartment syndromes, irrigation and debridement of wounds, application of external fixation devices, placement of intramedullary nails, possible limb amputations). The type and extent of open tibial fractures and soft tissue injuries determine the best treatment options for patients. Perioperative nurses should help patients focus on treatment choices for their open tibial fractures that ensure optimal surgical outcomes and maintain their quality of life. PMID- 8712810 TI - Management of upper abdominal solid organ injuries. AB - Solid organ injuries of the upper abdomen consist primarily of injuries to the spleen and liver. Several years ago, these injuries were fairly straightforward to manage. Recently, however, combinations of new diagnostic modalities and the advent of nonsurgical treatments have added complexity to the management of splenic and hepatic injuries. Splenic injuries that require surgical repairs are managed by splenic salvage when feasible and by splenectomy procedures when the injuries are complicated. Hepatic injuries may require a variety of therapeutic modalities, depending on their complexity. This article reviews some of these treatment options for the management of upper abdominal solid organ injuries. PMID- 8712812 TI - In search of perioperative nursing data elements. AB - A 14-member task force appointed by the AORN Board of Directors began the process of identifying perioperative nursing data elements in November 1993. During the ensuing two years, the task force members identified perioperative nursing intervention data elements, linked these data elements to specific patient outcomes, and asked 232 AORN members to validate the list of intervention data elements. This article explains the value of data elements to perioperative nurses, describes the consensus process used by the task force, and presents the Perioperative Nursing Data Elements: Interventions, Version 1.0. PMID- 8712811 TI - Traumatic rupture of the descending thoracic aorta. AB - Traumatic rupture of the descending thoracic aorta is a highly lethal injury caused by sudden, rapid deceleration in high-speed motor vehicular collisions and falls from heights. Deceleration causes the mobile ascending aorta and aortic arch to move forward while the descending thoracic aorta remains fixed by the mediastinal pleura. This opposing movement creates a tear at the aortic isthmus just distal to the origin of the left subclavian artery. Patients with traumatic ruptured descending thoracic aortas often have serious multisystem injuries, and approximately 85% of these patients die at accident scenes or during emergency department resuscitation. The most frequent cause of death is free rupture of periaortic tissues that temporarily have provided tamponade to the sites of aortic ruptures. Surgical mortality rates for patients who survive initial resuscitation are between 10% and 25%. Advances in field triage and emergency medical transportation systems and new preoperative evaluation modalities have improved the survival of patients with traumatic descending thoracic aorta ruptures. PMID- 8712813 TI - Perioperative care or elderly trauma patients. PMID- 8712814 TI - Recommended practices for selection and use of packaging systems. Association of Operating Room Nurses. PMID- 8712815 TI - [Telomerase]. AB - Telomerase is expected to be a powerful tool for cancer diagnosis and cancer chemotherapeutics because of its selective expression exclusively in various cancer tissues at a high frequency (90%) in human somatic tissues. Some precancerous tissues also express telomerase at a high frequency. Normal somatic tissues are mostly telomerase negative. Cancer cells die after a loss of telomerase activity. PMID- 8712816 TI - [Clinico-pathological predictive indicators in squamous cell carcinoma of the head and neck]. AB - The significance of prognostic factors that may predict the clinical outcome of patients with head and neck cancer was discussed. Many indicators can be grouped into three categories, patient factors, tumor factors and treatment factors. The most significant indicator of prognosis seems to be pathological nodal stage. Factors such as clinical stage, resectability, and depth of invasion may also affect the patient outcome. Recent research development has revealed biological phenotypes of cancer cells to predict the effect of cancer treatment and the clinical course in head and neck cancer. Possible predictive indicators include DNA ploidy, Tpot, EGFR and cyclin D1. C erbB2 and p53 may not predict the survival of patients with head and neck cancer. PMID- 8712817 TI - [Recent advance in head and neck cancer surgery]. AB - In order to remove a tumor radically, it is necessary to resect the tumor en-bloc together with surrounding normal tissues as much as possible. This results in a large tissue defect. Since the head and neck region has important functions for daily life such as respiration, speech, chewing, swallowing, the size of tissue defect directly affects post-operative functions and cosmetic results. Recently, the use of free flaps has made it possible to resect the advanced tumor which had been thought to be inoperable before. The introduction of free flaps has permitted reconstruction of the skull base, the nasopharynx, the mandible and the face. These were impossible to reconstruct with pedicled flaps such as delto pectoral flap and pectoral major musculo-cutaneous flap. Therefore, the indication of reconstructive surgery has been remarkably enlarged for advanced head and neck cancer. We employed reconstructive surgery using free flaps for head and neck cancer in 1982, and, in August 1994, the number of patients operated on with free flaps reached 1,000. Through our experience of head and neck reconstructive surgery, we can conclude that the use of free flaps for the reconstruction in the head and neck region has permitted enlarging surgical indication for far advanced head and neck cancers. And the free flaps have made it possible to assure oral functions such as speech and chewing after reconstruction of the palate or the mandible. The use of free flaps has also decreased the incidence of postoperative complications such as fistula, infection, etc. It is incontestable that reconstructive surgery is indispensable for the treatment of head and neck cancer. PMID- 8712818 TI - [Treatment of head and neck cancer: radiation therapy]. AB - Radiation therapy is an effective modality for curative treatment with organ preservation, palliative treatment for pain control and combined/prophylactic treatment for local-regional control after surgery in head and neck cancer. For early lesions, 70-90% of local control may be expected in radiation therapy. However, some hurdles must be cleared with regard to advanced lesions. We reviewed the prospective randomized controlled trials (phase III trial) concerning adjuvant chemotherapy and multiple fractions per day of radiotherapy for advanced head and neck cancer. Adjuvant chemotherapy may be used in three ways in the treatment, i.e., induction therapy, concomitantly with radiotherapy and maintenance therapy. We reviewed the former two treatments because no "pure" phase III trial could be found in maintenance therapy after radiotherapy. As for multiple fractions per day of radiation therapy, the following definition is used. Hyperfractionation: Total dose is increased; dose per fraction is reduced; fractian number is increased,; overall time is relatively unchanged. Accelerated fractionation: Overall time is reduced; number of fraction total dose, and dose per fraction are either unchanged or somewhat reduced, depending on the extent of overall time reduction. Adjuvant chemotherapy combined with radiotherapy and multiple fractions per day of radiotherapy have been attempted to increase local regional control and survival in a few randomized studies. However, these survival results were disappointing despite good local control. We need a more effective chemotherapy regime, fractionation schedule and also to clarify the indications for these treatments by analyzing the prognostic factors. PMID- 8712819 TI - [Chemotherapy of head and neck cancer]. AB - Squamous cell carcinoma of the head and neck is a chemotherapeutically responsive tumor. Methotrexate, cisplatin, 5-fluorouracil and bleomycin in various combinations have achieved tumor regression, with complete or partial response, in 60-90% of previously untreated patients. The response to chemotherapy, however, is often short-lived, and conventional treatment has produced little impact on survival. To date, neo-adjuvant and/or adjuvant chemotherapy have also failed to demonstrate any impact on the survival rate, although subset analysis has indicated a possible survival benefit. Furthermore, larynx preservation and reduction in incidence of metastasis have been demonstrated. Concomitant chemoradiotherapy has some positive results in disease free survival and/or overall survival. In this report, we reviewed the literature on neoadjuvant chemotherapy, adjuvant chemotherapy, salvage chemotherapy and concomitant chemoradiotherapy for advanced head and neck cancer, and analyzed their efficacy. PMID- 8712820 TI - [Immunotherapy of patients with head and neck carcinomas]. AB - The decline of immunity has been reported in head and neck carcinomas in the advanced stage. On the basis of this finding in head and neck carcinomas, additional immunotherapy in curative treatment, i.e., surgical therapy and/or radiation therapy, has been applied for patients with advanced tumors to improve their poor prognosis. Until now, only a few controlled randomized studies have evaluated the clinical efficacy of non-specific immunotherapy, i.e., Ubenimex and OK-432. Each result using these drugs showed that the survival rate in those receiving non-specific immunotherapy was higher than in the control. Cytokines, e.g., IFN-alpha, gamma, TNF-alpha, had immunologically modulating actions on the immunological conditions of patients and on the tumor cells, whereas immunotherapy using these cytokines has not shown a good clinical response, as with clinical administration of IL-2 alone. Furthermore, the clinical results of combined therapy of IL-2 and adoptive immunotherapy for head and neck carcinomas have not been satisfactory. In future, to improve the prognosis in patients with these carcinomas, it is indispensable to develop a new immunotherapy including transfer of cytokine-transfected cells having tumor-immunization ability and capable of inducing cytolytic effector cells in the peritumoral site, and specific antibody therapy against tumor cells. PMID- 8712821 TI - [Antitumor effects of Behenoyl-ara-C (BH-AC) in combination with Idarubicin (IDA) in P 388 leukemic cell bearing mice]. AB - The antitumor effects of Behenoyl-ara-C (BH-AC) in combination with Idarubicin (IDA) on leukemia were studied. First, a combination of IDA with Ara-C, which is the main metabolite of BH-AC, was evaluated with regard to its in vitro cytotoxic activity on mouse P 388 leukemic cells. The effect of this combination proved to be additive according to isobologram analysis. Secondly, the antitumor activity of an intravenous bolus-administration of a combination of BH-AC and IDA was evaluated by the life span of P 388 bearing mice, and compared with the activity of the Ara-C and IDA combination. The antitumor activity of Ara-C administered alone was clearly dependent on the administration schedule and was most intense when Ara-C was administered with the most frequent injections (3 bolus injections/day x 3 days), whereas antitumor activity of BH-AC was less dependent on the schedule. IDA administered alone showed dose-dependency in its antitumor activity up to 3 mg/kg. The maximum effects of IDA were observed with amounts of 3 - 4 mg/kg. In the same leukemia model, the combination of frequent injections of BH-AC and a single injection of IDA (increased life span: ILS>300%; cure ratio: CR = 3/5) conferred a more potent effect compared to the results of BH-AC (ILS = 133%, CR = 0/5) or IDA (ILS = 67%, CR = 2/5) alone. The effect of BH-AC and IDA combination was comparable or superior to that of the Ara-C and IDA (ILS = 233%, CR = 2/5) combination. These results indicated the possibility of clinical usefulness with a combination therapy of BH-AC and IDA against leukemia. PMID- 8712822 TI - [Effects of carboplatin combination with etoposide against leukemia/lymphoma cell lines]. AB - We studied the effects of carboplatin in combination with etoposide in human B cell lymphoma cell lines, BALL-2, Dauji and human T-cell leukemia cell lines, CEM, HSB and MOLT-3 cells. Cells were incubated for 3 days in the presence of carboplatin and etoposide, and the combined drug and cell growth inhibition was determined by MTT assay. The effects of drug combinations at ID80 were analyzed by an improved isobologram method (Steel and Peckham). In the combination of carboplatin with etoposide, the data points fell in the envelope of additivity (additive effect) in all five cell lines. Synergistic and antagonistic effects were not observed. These findings suggest that the combination of carboplatin and etoposide are as effective as expected. PMID- 8712824 TI - [The indications for postoperative adjuvant therapy in node-negative breast cancer patients]. AB - A clinicopathological study on prognosis has been carried out in 233 breast cancer patients with more than twenty inspected nodes and without lymph node metastasis. Multivariate analysis of clinicopathological findings in node negative breast cancers showed that the best combination of clinical features predicting prognosis were age, menstruation status, tumor location and tumor diameter; and from 11 pathological features, three factors (histological type, histological grade and the grade of tumor infiltrating lymphocytes) were selected under condition of p<0.05. With solid-tubular carcinoma, histological grade III and negative tumor infiltrating lymphocytes as the 3 poor prognostic factors, node-negative breast cancers were classified. The recurrence rates of subgroups with the number of 3, 2, 1 and 0 were 28%, 16%, 7% and 0%, respectively. We conclude that the grade of tumor infiltrating lymphocytes is especially significant as the prognostic factor in node-negative breast cancers, and that postoperative adjuvant chemoendocrine therapy must be performed for patients with node-negative breast cancer with more than two poor prognostic pathological factors. PMID- 8712823 TI - [A randomized controlled study of post-operative adjuvant therapy in non-small cell lung cancer]. AB - The efficacy of postoperative adjuvant chemo- and chemoimmunotherapy in non-small cell lung cancer was evaluated in a multicentric prospective randomized study. From September 1987 to June 1990, resected lung cancer patients were randomly stratified into three groups. Group A received 2 courses of chemotherapy with CDDP and VDS following operation. Group B was administered UFT daily for 1 year after 2 courses of CDDP. Group C received intrapleural administration of OK-432 after lung resection, then UFT and OK-432 once every 2 weeks for 1 year. Out of 94 cases, analyses were carried out on 87 of eligible cases. The five-year survival rate was 56.8% in stage I (43 cases), 73.3% in stage II (12 cases), 18.8% in stage IIIA (24 cases), 50% in stage IIIB (2 cases) and 33.3% in stage IV (6 cases). The five-year survival rate in group A was 32.2%, 55.2% in group B and 53.9% in group C, and no statistical difference was recognized between 3 groups. But in the cases of noncurative resection, the 5-year survival rate was significantly low in group A compared with Group B or C. Similarly, the cases with low-grade TP (<6.0 g/dl) or low response of PPD skin reaction (< 12mm) showed a significantly low 5-year survival rate only in group A. From these results, it was suspected that aggressive chemotherapy provides no benefit for postoperative lung cancer patients with advanced disease. PMID- 8712825 TI - [Phase II study of paclitaxel (BMS-181339) in patients with ovarian cancer by 3 hour intravenous infusion]. AB - A phase II study of Paclitaxel in patients with ovarian cancer by 3-hour intravenous infusion was undertaken by a cooperative study group of 30 institutes. Of 66 cases enrolled, 57 cases were evaluable for efficacy, and 63 cases were evaluable for safety. In spite of the fact that all cases for efficacy evaluation were previously treated with chemotherapy including platinum-based drugs, 2 cases of complete response (CR) and 15 cases of partial response (PR) were observed, with a response rate of 29.8% (The 95% confidence interval of response rate was 18.4-43.4%). Paclitaxel also showed 28.2% (11/39) response rate in patients refractory to treatment by platinum-based drugs. Histologically, the response rates were 28.9% (11/38) in serous adenocarcinoma, 40.0% (2/5) in clear cell adenocarcinoma and 25.0% (1/4) in mucinous adenocarcinoma. As the major laboratory abnormalities, leukopenia, neutropenia and decrease in hemoglobin were observed with incidence rates of 98.4% (62/63), 95.2% (59/62) and 85.7% (54/63), respectively. However, these abnormalities were clinically manageable by either withdrawal of medication, administration of antibiotics, G-CSF or metachysis etc. In addition, thrombocytopenia, elevation in GOT and GPT were seen with moderate incidence. Peripheral neuropathy was a major adverse symptom with an incidence of 79.4% (50/63), followed by alopecia, myalgia, arthralgia and fever. However, the majority of these adverse reactions were less than grade 3. From these findings, we confirmed that 3-hour intravenous infusion of Paclitaxel was a clinically useful chemotherapeutic agent in patients with ovarian cancer. PMID- 8712826 TI - [Randomized controlled study on adjuvant immunochemotherapy with carmofur (HCFU) for noncuratively resected and unresected gastric cancer]. AB - To evaluate the efficacy of adjuvant immunochemotherapy, especially the long-term administration of HCFU to patients with noncuratively resected and unresected gastric cancer, a randomized controlled study was conducted by 16 institutions in Hokkaido. After surgery, 185 patients were divided into two groups using the envelope method; group A (84 patients) treated by MMC (4 mg, twice a week, total 1 mg/kg)+OK-432 (or PSK): group B (101 patients) treated with MMC+OK-432+HCFU (600 mg, per day). The 5- and 10-year survival rates were 9.8 and 3.1% in group A, and 11.1 and 11.1% in group B (p=0.062). Better survival rates were obtained in those cases with unresected, S3, or peritoneal dissemination in group B. These results suggested the long-term administration of HCFU was effective for noncuratively resected and unresected gastric cancer. PMID- 8712827 TI - [Development of quality of life questionnaire for patients with colorectal cancer in surgical area--a study of reliability and validity of Tokyo Yamabuki Forum Version]. AB - We developed a new questionnaire in the surgical area based on a core quality of life (QOL) questionnaire for patients with gastrointestinal cancer. In this study, we investigated the validity and reliability of a QOL questionnaire (Tokyo Yamabuki Forum Version) for patients with colorectal cancer. The questionnaire was composed of 17 items including 5 scales (basic sensory scale, psychological scale, physiological scale, defection-related scale and active scale) and a face scale as an global scale. The time needed to answer questionnaires was expected to be around 7 minutes and the questionnaires should basically be answered by the patients themselves everyday in the hospital. The study was performed in 10 hospitals in the Tokyo area, and 394 samples collected from 21 patients with rectal and colonic cancers were analyzed. A number of respondents failed to answer the question "Do you feel your foods tasty?", so we judged this item inappropriate and deleted it from the analysis. Fifteen items, including 5 scales showed satisfactory internal consistency and construct validity in correlation and factor analyses. Performance status showed a low correlation between each item, each scale and the global scale, while SDS and STAI showed an inordinately negative correlation with the fundamental and physical scales. Especially, SDS revealed an extremely close correlation with the active scale, and STAI showed an excessive correlation with the psychological scale. In the time course of QOL under chemotherapy, reductions (aggravations) were observed in both the total score of 15 items and global scale within one week postoperatively, but after that recovered to preoperative levels at 2 weeks postoperatively. A tendency to QOL improvement was observed 2 weeks after starting chemotherapy or chemoimmunotherapy. QOL of 13 patients was measured over 3 months, and the longest term was 8 months. The results suggested that this QOL questionnaire has sufficient reliability and validity to be usable for patients with colorectal cancer in the surgical area and that this model is applicable for long-term QOL surveys and frequent measurement. PMID- 8712828 TI - [Somatostatin analogue treatment for malignant hypercalcemia associated with advanced breast cancer]. AB - We describe the successful treatment with octreotide, a somatostatin analogue, of a patient with malignant hypercalcemia associated with advanced breast cancer. A 70-year-old female with advanced breast cancer was admitted to our department for treatment of hypercalcemia. The administration of pamidronate disodium was effective to decrease serum calcium from 6.2 mEq/l to 4.0 mEq/l for the first time, but her hypercalcemia later responded less to pamidronate, and her serum calcium remained raised in spite of the administration of pamidronate and elcatonin. Then, her condition deteriorated with hypercalcemic symptoms, such as nausea vomiting and drowsiness. After octreotide treatment (100 microg/body/day, s.c.) with a combination of prednisolone, her serum calcium level improved from 6.7 mEq/l to 5.0-5.5 mEq/l, Leading to a dramatic improvement in her symptoms. During these treatments, anti-cancer therapy, hydration and the administration of diuretics have been continued. We think octreotide is very useful for the treatment of malignant hypercalcemia associated with advanced breast cancer. PMID- 8712829 TI - [A case of carcinomatous peritonitis for which combination therapy of 5-FU, leucovorin and cisplatin was effective]. AB - This case report describes a case of carcinomatous peritonitis which showed a good response to the combination therapy of 5-FU, leucovorin and cisplatin. A 55 year-old man was admitted with complaints of meteorism due to ascites fluid. Ascitic examination showed a high concentration of CEA and CA19-9, and adenocarcinoma cells. Laboratory and radiological examination did not reveal the origin of the carcinoma, and laparotomy suggested an appendiceal origin. The combination chemotherapy with 5-FU, leucovorin and cisplatin was conducted for a total of 11 courses for 2 years with good QOL until his death. This therapy was suggested to be useful for inoperable carcinomatous peritonitis. PMID- 8712830 TI - [A case of advanced gastric cancer remarkably responding to preoperative UFT-E therapy]. AB - We here reported a case of advanced gastric cancer remarkably responding to preoperative short-term UFT-E chemotherapy. UFT-E was orally administered preoperatively for about a month to the patient with type 2 advanced gastric cancer. After the chemotherapy the cancer was found to be remarkably decreased in size and denatured. The amount of residual cancer cells was limited by histopathological examination following the operation and diagnosed as Grade 3 based on the criteria of histological evaluation of chemotherapy for cancer. We continued to administer UFT-E postoperatively and the patient is still alive without symptoms. PMID- 8712831 TI - [Three cases of drug-induced hemolytic uremic syndrome]. AB - We report three cases of drug-induced hemolytic uremic syndrome (HUS). Three patients with advanced gastrointestinal cancer underwent a curative operation and adjuvant chemotherapy with Mitomycin C (MMC), 5FU and Ara-C. Later, progressive anemia, thrombocytopenia, renal dysfunction and elevation of serum LDH were recognized. A diagnosis of HUS was made. As they had no symptoms of infectious diseases or relapse of cancer, the cause of HUS was thought to be MMC. Treatment with antiplatelet drugs and fresh frozen plasma was effective for two patients. However, one patient died of pulmonary edema. PMID- 8712832 TI - [Rhabdomyosarcoma of the bladder: a case report]. AB - A 15-year-old man with the chief complaint of general fatigue was referred to our hospital on November 11, 1993. Bilateral percutaneous nephrostomy was performed for postrenal anuria. X-ray examinations revealed a huge intrapelvic tumor, and it was histopathologically diagnosed as rhabdomyosarcoma by transrectal needle biopsy. He was initially treated with combination chemotherapy regimen of vincristine, actinomycin-D and cyclophosphamide (VAC therapy). Pelvic exenteration was performed on December 15, 1993. Histopathological findings were alveolar rhabdomyosarcoma with degenerative change and partial necrosis. After the operation, he was given two course of VAC therapy. In May, 1994, brain metastasis occurred, so 4 courses of VAC therapy were administered. For a very short period, neurological symptoms improved, but he died of pneumonia on November 15, 1994. PMID- 8712833 TI - [Neoadjuvant therapy followed by administration of UFT for breast cancer patients]. PMID- 8712834 TI - [Fourth Early Breast Cancer Trialists' Collaborative Group meeting in Oxford- meta-analysis of early breast cancer clinical trials]. PMID- 8712835 TI - [Nedaplatin]. AB - Nedaplatin is a derivative of cisplatin which produced less nausea & vomiting and nephrotoxicity. In the phase I study, the MTD was 120 mg/m2 and the DLF was a bone marrow suppression. The optimal dose in a phase II study was judged to be 100 mg/m2 repeated every 4 weeks. In the phase II studies, response rates obtained were 42.2% for head & neck ca., 40.9% for small cell lung ca. (SCLC), 20.5% for non-SCLS (NSCLC), 12.5% for breast ca., 51.7% for esophageal ca., 8.3% for stomach cancer. 0 for colon ca., 38.1% for bladder ca., 14.3% for pyelo ureter tract ca., 18.8% for prostatic ca., 80.0% for testicular tumor, 37.3% for ovarian ca., 46.3% for cervical ca. Grade 3.4 thrombocytopenia, leukopenia, anemia and nausea & vomiting were found in 28.5%, 21.1%, 16.8% and 18.5% respectively. In an additional phase II study for cervical ca. at a dose reduced to 80 mg/m2, a response rate was comparable together with less thrombocytopenia. In a randomized controlled study of nedaplatin plus vindesine vs. cisplatin plus vindesine in NSCLC, there was no significant difference in response, however mephro and G.I. toxicity were significantly less in the nedaplatin group. Thrombocytopenia was found more frequently in the nedaplatin groups. Based on the results, the indication was approved in ca. of the head & neck, SCLC, NSCLC, esophagus, bladder, testicular tumor, ovary and cervix. Dose schedule is 80 - 100 mg/m2 every 4 weeks at more 1,000 mL drip infusion repeated. PMID- 8712836 TI - Early detection of melanoma metastasis using fludeoxyglucose F 18 positron emission tomography. PMID- 8712837 TI - The melanoma epidemic. Is increased surveillance the solution or the problem? AB - Melanoma has a very bad reputation, and rightfully so. Until relatively recently, the prognosis for melanoma was dismal and, very reasonably, early detection has been proposed as a viable approach to preventing melanoma deaths. On the surface, this simple approach appears to have been a stellar success, with melanoma survival rates improving markedly. There is no question that there has been a dramatic increase in the number of people with a diagnosis of melanoma. On careful scrutiny, the numbers reflected in the increased incidence may not add up. Mortality rates and the diagnosis rate for thick tumors ( > 1.5 mm thick) have not gone up commensurate with the growing rate of diagnosis. The obvious explanation for this discrepancy is that we are doing a great job! However, this explanation must assume that an overwhelming percentage of those at risk for melanoma are in a position to benefit from our service. It is very unlikely that this is the case. It is not clear whether the increase in the number of melanoma cases diagnosed reflects an actual increase in real disease. Could this increase be in part a product of aggressive surveillance resulting in the identification of "atypical" pigmented skin tumors of limited or nonexistent potential for malignant behavior? Could the dramatic improvement in the prognosis of melanoma simply be a consequence of removing biologically benign pigmented tumors that are inadvertently classified as malignant? PMID- 8712838 TI - From bedside to the bench and back. Nitric oxide and the cutis. AB - When the discoverer of dynamite (trinitrotoluene [TNT]), Alfred Nobel, was prescribed nitroglycerin for angina in 1895, he was certainly taken aback. Almost a century later, organic nitrates and their gaseous metabolic end product, nitric oxide (NO), were implicated in a vast array of biologically diverse activities. About 10 years ago, a series of discoveries from different avenues of research converged on NO, thrusting it into the limelight as a neurotransmitter, vasodilator, toxin, and modulator of immune function and inflammation. Nitric oxide has thus managed to capture the interest of scientists from a number of fields and holds center stage attention. Interest in NO among dermatologists has been slow to appear, however, and the literature on NO with respect to the skin is sparse when compared with the steep escalation in the number of articles published generally on NO since 1987 (Figure 1). PMID- 8712839 TI - Comparative percutaneous absorption of lindane and permethrin. AB - BACKGROUND AND DESIGN: Because of the concern for potential neurotoxic effects (central nervous system excitation, convulsions) in the treatment of scabies using 1% lindane lotion, 5% permethrin cream has been suggested as an alternative scabicide. Using the finite dose technique, in vitro percutaneous absorption of 5% permethrin cream or 1% lindane lotion was measured in human and guinea pig skin following a single application. In vivo blood and brain levels of the scabicides were measured in guinea pigs following three daily applications of 5% permethrin cream or 1% lindane lotion. Permethrin and lindane levels were quantified by gas chromatography/mass spectroscopy. RESULTS: In vitro percutaneous absorption of the two scabicides was identical in guinea pig skin; however, human skin was 20-fold more permeable to lindane than to permethrin. In vivo guinea pig blood and brain levels of lindane were fourfold greater than permethrin levels. CONCLUSION: The risk for toxic effects, as assessed by systemic exposure during overuse conditions, is projected to be 40 to 400 times lower for 5% permethrin cream than for 1% lindane lotion. PMID- 8712841 TI - Deposition of granular IgA relative to clinical lesions in dermatitis herpetiformis. AB - OBJECTIVE: To compare the deposition of IgA and C3 in the skin of patients with active dermatitis herpetiformis relative to the sites of disease. DESIGN: In the phase 1 study, skin biopsy specimens were obtained from erythematous perilesional skin, nonerythematous perilesional skin, and never-involved skin. In the phase 2 study, specimens from the nonerythematous perilesional and uninvolved skin from the same anatomic region were sampled. SETTING: The Dermatology Clinic at the University of Utah Health Sciences Center, Salt Lake City. PATIENTS: Patients with known dermatitis herpetiformis: 19 patients in the phase 1 study and 15 patients in the phase 2 study. Suppressive medications were stopped for 48 to 72 hours after biopsy specimens were obtained. All patients had active disease at the time that biopsy specimens were taken. MAIN OUTCOME MEASURE: The intensity of IgA and C3 immunofluorescent staining in 6 sections from each skin biopsy specimen was graded by using a semiquantitative scale (0 to 3+) in a blinded fashion by a single observer. RESULTS: Deposition of IgA was more intense in noninflamed perilesional skin in 11 of 19 patients compared with that in erythematous skin (P < .05). Erythematous skin was negative for IgA in 16% (3/19) of the specimens. Noninflamed perilesional skin showed more intense IgA deposition in 18 of 19 specimens compared with that in never-involved skin (P < .01); C3 was more intense in erythematous skin (P < .01). In the phase 2 study, skin from the same anatomic region revealed greater deposition of IgA near lesions in 12 of 15 patients (P < .001). CONCLUSIONS: In patients with dermatitis herpetiformis, IgA is not uniformly distributed throughout the skin, and IgA is present in greater amounts near active lesions. The preferred biopsy site for the diagnosis of dermatitis herpetiformis is normal-appearing skin that is adjacent to an active lesion. PMID- 8712840 TI - Abnormal serum porphyrin levels in patients with the acquired immunodeficiency syndrome with or without hepatitis C virus infection. AB - OBJECTIVE: To define the contributions of human immunodeficiency virus (HIV) and hepatitis C virus infections to the development of porphyria cutanea tarda. DESIGN: Analysis of serum porphyrin levels in a cohort of 167 subjects. Serum samples were divided into 4 groups corresponding to the status of HIV and hepatitis C virus infections: positive-positive, positive-negative, negative positive, and negative-negative. SETTING: Serum samples positive for HIV were obtained from the serum bank of an acquired immunodeficiency syndrome-HIV research center, and HIV-negative samples were obtained from a regional blood center. MAIN OUTCOME MEASURES: Spectrofluorometric measurement of serum porphyrin levels. RESULTS: The median values of porphyrin were 2.31 nmol/L (interquartile range [difference between the 25th and 75th percentiles]: 4.55) in the positive positive group, 1.99 nmol/L (interquartile range: 1.63) in the positive-negative group, 1.31 nmol/L (interquartile range: 1.58) in the negative-positive group, and 1.14 nmol/L (interquartile range: 0.92) in the negative-negative group. The fluorescence emission spectra of samples with elevated porphyrin levels were identical with that reported for porphyria cutanea tarda. Elevated porphyrin levels were significantly associated with HIV infection (P < .001) and were observed in patients with an elevated level of alanine aminotransferase (P = .03). Infection with hepatitis C virus was also associated with an elevation in porphyrin levels, although the increase was not statistically significant (P = .16). Porphyrin levels in patients positive for HIV were not correlated with serum urea nitrogen or creatinine levels. None of the patients had symptomatic porphyria cutanea tarda. CONCLUSIONS: Factors associated with increased serum porphyrin levels included HIV infection, elevated alanine aminotransferase levels, and, to a lesser extent, hepatitis C virus infection. These findings suggest that patients with the above risk factors are potentially predisposed to the development of symptomatic porphyria cutanea tarda on further exposure to hepatotoxic agents. PMID- 8712843 TI - Metastatic Crohn's disease. Report of 3 cases with special reference to histopathologic findings. AB - BACKGROUND: Metastatic Crohn's disease is a rare, often unrecognized cutaneous disorder lacking definite histopathologic criteria. The purpose of this study was to document clinicopathologic and immunologic findings in 3 patients with metastatic Crohn's disease. The histopathologic findings are evaluated in correlation to those reported in the literature in an attempt to better define the histopathologic features. OBSERVATIONS: None of the patients showed signs of depressed cell-mediated immune response as evaluated with skin tests and T-cell subtyping of blood samples. One of the patients had antineutrophil cytoplasmic antibodies. Polymerase chain reaction, a highly efficient method of amplifying low levels of specific DNA sequences, did not show mycobacterial DNA in the samples studied. Granulomas of the sarcoid type with numerous foreign body and Langhans giant cells were the dominating features. In accordance with previous results, we found vascular involvement in 2 cases, manifested as granulomatous perivasculitis in both. We also found necrobiotic areas in all 3 cases. CONCLUSIONS: We propose that both necrobiosis and granulomatous perivasculitis be added to the histopathologic characteristics of metastatic Crohn's disease. Patients may even have a positive antineutrophil cytoplasmic antibody test result. PMID- 8712842 TI - Absence of detectable alpha 6 integrin in pyloric atresia-junctional epidermolysis bullosa syndrome. Application for prenatal diagnosis in a family at risk for recurrence. AB - BACKGROUND AND DESIGN: The expression of basement membrane-related antigens was surveyed in 2 Japanese siblings who died of pyloric atresia-junctional epidermolysis bullosa syndrome in early infancy. RESULTS: The skin specimens of both patients demonstrated complete absence of detectable alpha 6 integrin and markedly reduced amounts of beta 4 integrin. All the other subtypes of epidermolysis bullosa used as controls demonstrated normal intensity of expression of alpha 6 and beta 4 integrin. In contrast to the negative immunoreactivity of monoclonal antibody GB3 in gravis-Herlitz junctional epidermolysis bullosa (n = 4), a bright linear pattern along the epidermal basement, membrane was demonstrated in the skin of both siblings with pyloric atresia-junctional epidermolysis bullosa syndrome. Based on these data, a monoclonal antibody against alpha 6 integrin was successfully used as a prenatal diagnostic probe for a skin biopsy specimen from a fetus at risk for pyloric atresia-junctional epidermolysis bullosa syndrome in this family. CONCLUSION: The absence of detectable alpha 6 integrin, but not beta 4 integrin, in these cases raises the possibility that alpha 6 integrin or its ligands are responsible for the pyloric atresia-junctional epidermolysis bullosa syndrome phenotype seen in this family. PMID- 8712844 TI - Serious adverse reactions induced by minocycline. Report of 13 patients and review of the literature. AB - BACKGROUND: Minocycline has been reported to cause serious, albeit rare, adverse events, including serum sickness-like reaction, hypersensitivity syndrome reaction, and drug-induced lupus. A retrospective review of patients seen in our Adverse Drug Reaction Clinic as well as information obtained from the Health Protection Branch was done to identify patients with minocycline-induced reactions. In addition, the literature concerning serious reactions to minocycline was reviewed. OBSERVATIONS: Six patients with a hypersensitivity syndrome reaction, 6 patients with a serum sickness-like reaction, and 1 patient who had symptoms consistent with drug-induced lupus were identified. A review of the literature identified 11 cases of hypersensitivity syndrome reaction, 1 case of serum sickness-like reaction, and 24 cases of drug-induced lupus. Serum sickness-like reactions occur sooner than hypersensitivity syndrome reactions (15.6 vs 23.7 days, P = .04). Drug-induced lupus occurs on average 2 years after the start of minocycline therapy. CONCLUSIONS: Dermatologists need to be aware of the serious adverse reactions that can develop after minocycline use. In patients who may require long-term therapy with minocycline ( > 1 year), we suggest that antinuclear antibody and hepatic transaminase levels be determined at baseline. Rechallenge with minocycline or other tetracyclines is currently not recommended for patients who develop these serious reactions. PMID- 8712845 TI - Learning and teaching in dermatology. A practitioner's guide. AB - BACKGROUND: The explosion of medical knowledge, the influence of communications technology, and the pressures for health care reform challenge dermatologists to learn efficiently and teach effectively. OBSERVATIONS: The vast body of material on teaching and learning in the education literature contrasts sharply with the limited information on these topics in the dermatology literature. Gaining an understanding of these approaches to learning provides a foundation for the design of personal continuing medical education plans and lesson plans for students. Practical teaching skills are emphasized in this review. CONCLUSIONS: Learning and teaching skills can be improved through study. Successful teaching often produces positive feedback, which can lead to renewed enthusiasm for educating clinicians in the field of dermatology. PMID- 8712846 TI - What every dermatologist should know about homeopathy. PMID- 8712847 TI - Safety of permethrin vs lindane for the treatment of scabies. PMID- 8712848 TI - Disseminated ulcerated papules and nodules. Cutaneous disseminated sporotrichosis. PMID- 8712849 TI - Vesicular eruption on the scalp. Varicella-zoster virus with Torulopsis glabrata colonization. PMID- 8712850 TI - A palmar chancre and multiple proximal erythematous nodules. Nodular lymphangitis due to Nocardia brasiliensis. PMID- 8712851 TI - Widespread annular erythematous plaques. Multibacillary leprosy. PMID- 8712852 TI - Mycosis fungoides palmaris et plantaris. PMID- 8712853 TI - Possible neural basis for the field effect in local recurrence of melanoma in situ. PMID- 8712854 TI - Classification of cutaneous lymphomas. PMID- 8712855 TI - Detection of antineutrophil cytoplasmic antibodies in patients with hepatitis C virus-induced cutaneous vasculitis with mixed cryoglobulinemia. PMID- 8712856 TI - Papular-purpuric 'gloves and socks' syndrome. PMID- 8712857 TI - Disseminated strongyloidiasis in a patient with acquired immunodeficiency syndrome. PMID- 8712858 TI - Successful treatment of Sezary syndrome with lymphomatous transformation to large cell lymphoma with fludarabine phosphate. PMID- 8712859 TI - Spiking of the tibial tubercles--a radiological feature of osteoarthritis? AB - OBJECTIVE: To determine whether 'spiking' or angulation of the tibial tubercle is associated with other radiographic markers of osteoarthritis (OA) or pain in the knee joint, and could be taken as a reliable marker for early OA, in a large general population sample. METHODS: A total of 950 women from the Chingford general population survey underwent anteroposterior extended weight bearing radiography of the knees. Angulation of the tip of the medial and lateral tubercles, and height of the tubercles above the tibial plateau were measured. These measures were compared with standard radiographic indices including qualitative Kellgren and Lawrence global score, individual scores of osteophytes and joint space narrowing, and pain score. Intraobserver and interobserver reproducibility for assessment of spiking was tested in a subgroup of 50 films using two observers and two readings. Tibial spiking (angulation and height) was defined for this study as the top 10th centile for the whole population. Patients with normal radiographs (Kellgren and Lawrence grade 0) were allocated to quartile groups on the basis of spiking to define severity. Odds ratios were then calculated for the association of spiking and knee pain. RESULTS: The majority of the measures of tibial spiking were highly reproducible. There was a significant correlation between tibial spike angulation and the presence of osteophytes, but not joint space narrowing. The correlations for spike height with osteophytes and joint space narrowing were poor. There was an association between spike angulation at the lateral tubercle and reported knee pain (odds ratio 1.45 (95% confidence interval 1.03 to 2.03)) after adjustment for age, body mass index, and Kellgren and Lawrence score. There was no association between medial spike angulation or spike height and pain. Among the 950 women, 683 (72%) had normal radiographs (Kellgren and Lawrence = 0); in this group there was a similar association between pain and lateral spike angulation, but not medial spike angulation or spike height. CONCLUSIONS: Tibial spiking is associated with the presence of knee osteophytes and is reproducible, but does not have a strong independent relationship with knee pain. In patients with normal radiographs there is no useful correlation between tibial spiking and pain. Isolated tibial spiking is not a reliable sign of early knee OA, and should not routinely be reported. PMID- 8712860 TI - Prevalence and possible pathological significance of calcium phosphate salt accumulation in tendon matrix degeneration. AB - OBJECTIVES: To investigate the prevalence of calcium phosphate mineral salt accumulation in degenerative supraspinatus 'tendinitis' compared with a normal sample of human tendons, and to determine whether there is an association of calcium salt deposition with pathological changes in the tendon extracellular matrix. METHODS: Cadaver tendons (supraspinatus and common biceps tendons, n = 96) and fragments of supraspinatus tendons obtained during shoulder surgery (n = 31) were analysed for calcium content by atomic absorption spectroscopy, phosphorous content using a spectrophotometric assay, and matrix composition (collagen, glycosaminoglycans and DNA) using standard biochemical techniques. RESULTS: We established baseline values of calcium concentration in macroscopically normal cadaver tendons (mean 1.1 (SD 0.35) micrograms/mg dry wt, n = 60) and found that 33% (nine of 27) of ruptured tendons from patients with 'degenerative tendinitis' contained an excess of calcium (more than 2SD greater than the normal sample mean). Five of these specimens had increased concentrations of phosphorous and calcium:phosphorous (molar) ratios consistent with a variety of possible calcium crystals, including calcium pyrophosphate, hydroxyapatite, and tricalcium phosphate, in addition to mixed or amorphous calcium phosphate deposits. Four of these specimens contained normal concentrations of phosphorous, consistent with deposits of calcium oxalate or calcium carbonate, although this was not confirmed biochemically. In contrast, surgical specimens (n = 4) from patients with 'calcifying tendinitis' (radiographically detected calcium deposits) all contained salts with a mineral composition consistent with hydroxyapatite. The presence and identity of crystal deposits was subsequently confirmed in five specimens by radiographic microanalysis. Analysis of the tendon matrix demonstrated a number of significant differences between normal and degenerate (ruptured) tendons, including a reduction in collagen content, an increase in sulphated glycosaminoglycans (predominantly dermatan sulphate) and an increase in DNA (cellular) content. However, there were no significant differences between degenerate tendons that were 'calcified' and those degenerate specimens that contained normal concentrations of calcium. CONCLUSIONS: Although there was a relatively high prevalence of calcium salts in degenerate tendons, which might contribute to the pathological process (such as increased matrix collagen degradation), these data are consistent with the hypothesis that 'dystrophic calcification' of degenerate tendon matrix is a pathological entity distinct from cell mediated 'calcifying tendinitis'. Calcification is probably one possible outcome (or end point) of chronic tendon injury, although the possibility exists that in many cases, the presence of calcium salts may contribute to the tendon matrix degeneration. PMID- 8712861 TI - Assessment of the European classification criteria for Sjogren's syndrome in a series of clinically defined cases: results of a prospective multicentre study. The European Study Group on Diagnostic Criteria for Sjogren's Syndrome. AB - OBJECTIVE: To assess the recently proposed preliminary criteria for the classification of Sjogren's syndrome (SS) in a multicentre European study of a new series of clinically defined cases. METHODS: The criteria included six items: I = ocular symptoms; II = oral symptoms; III = evidence of keratoconjunctivitis sicca; IV = focal sialoadenitis by minor salivary gland biopsy; V = instrumental evidence of salivary gland involvement; VI = presence of autoantibodies. Each centre was asked to provide five patients with primary SS, five with secondary SS, five with connective tissue diseases (CTD) but without SS, and five controls (patients with ocular or oral features that may simulate SS). The preliminary six item classification criteria set was applied to both the SS patients and the non SS controls, and the performance of the criteria in terms of sensitivity and specificity was tested. RESULTS: The criteria set was tested on a total of 278 cases (157 SS patients and 121 non-SS controls) collected from 16 centres in 10 countries. At least four of the six items in the criteria set (limiting item VI to the presence of Ro(SS-A) or La(SS-B) antibodies) were present in 79 of 81 patients initially classified as having primary SS (sensitivity 97.5%), but in only seven of 121 non-SS controls (specificity 94.2%). When the presence of item I or II plus any two of items III-V of the criteria set was considered as indicative of secondary SS, 97.3% (71 of 73) of the patients initially defined as having this disorder and 91.8% (45 of 49) of the control patients with CTD without SS were correctly classified. CONCLUSION: This prospective study confirmed the high validity and reliability of the classification criteria for SS recently proposed by the European Community Study Group. PMID- 8712862 TI - Von Willebrand factor, thrombomodulin, thromboxane, beta-thromboglobulin and markers of fibrinolysis in primary Raynaud's phenomenon and systemic sclerosis. AB - OBJECTIVE: To determine whether measurement of different markers of endothelial damage, activation of coagulation, and platelet activation might differentiate between patients with primary Raynaud's phenomenon (PRP), limited cutaneous and diffuse systemic sclerosis (lcSSc and dSSc), and healthy control subjects. METHODS: Under carefully controlled conditions, fasting blood was drawn from 19 healthy control subjects, 10 patients with PRP, 17 with lcSSc and nine with dSSc for measurement of the following: von Willebrand factor (VWF) and soluble thrombomodulin as markers of endothelial damage/activation, thromboxane (as thromboxane B2) and beta-thromboglobulin as markers of platelet activation, and tissue plasminogen activator antigen, tissue plasminogen activator activity and plasminogen activator inhibitor-1 (PAI-1) as markers of fibrinolysis. RESULTS: VWF was increased significantly in patients with SSc, and there was also a linear trend for thromboxane and tissue plasminogen activator antigen (in addition to VWF) to differentiate between different subgroups of patients with Raynaud's phenomenon. Patients with dSSc had the highest values. A combined index of VWF and thromboxane showed a highly significant trend across the four groups studied. CONCLUSION: VWF, and to a lesser extent thromboxane and tissue plasminogen activator antigen, are associated with disease severity in patients with Raynaud's phenomenon. Prospective studies are now required to establish if these parameters can be used as markers of disease progression. PMID- 8712863 TI - Neutrophil adhesion to endothelial cells and factors affecting adhesion in patients with Behcet's disease. AB - OBJECTIVES: To study the in vitro adhesion of polymorphonuclear leucocytes (PMNLs) to endothelial cells in patients with Behcet's disease (BD), and the humoral and cellular factors which may contribute to adhesion. METHODS: A total of 118 patients with BD and 60 healthy controls were studied. In vitro adhesion of chromium-51 labelled normal neutrophils to human umbilical vascular endothelial cell (HUVEC) monolayers were studied in the presence of normal serum or serum obtained from patients with BD. Adhesion of neutrophils from patients with BD to HUVEC stimulated with tumour necrosis factor (TNF), interleukin-1 (IL 1), and lipopolysaccharide (LPS) and adhesion molecule (CD11a, CD11b, CD18 and L selectin) expression on the patient's neutrophils and lymphocytes were determined, and the serum concentration of IL-8 was measured. RESULTS: Sera from patients with BD were found to enhance the adherence of normal PMNLs to HUVEC monolayers in vitro. Patients' sera induced an increase in surface expression of CD11a and CD18 on normal neutrophils and intercellular adhesion molecule-1 (ICAM 1) expression on HUVECs. The number of CD11a positive neutrophils was greater in the blood of patients with BD than in that of healthy controls (89.4% v 71%; p < 0.001). Pretreatment of HUVECs with IL-1 alpha, TNF alpha or LPS resulted in an increased adhesion of patients' PMNLs greater than that observed for normal PMNLs. Monoclonal antibodies to CD11a, CD11b, CD18, and ICAM-1 caused varying degrees of inhibition of neutrophil adhesion. The concentration of IL-8 was also found to be significantly increased in sera of patients with BD (490 (SD 470) pg/ml) compared with normal controls (97.5 (56.3) pg/ml). CONCLUSION: Abnormalities of neutrophils, endothelial cells, or both, have been suggested to be responsible for many of the clinical manifestations of BD. Our findings may explain the underlying mechanism of neutrophil accumulation in Behcet's lesions. PMID- 8712864 TI - Choosing the best method for radiological assessment of patellofemoral osteoarthritis. AB - OBJECTIVE: To assess the reproducibility of different methods of radiological assessment of patellofemoral osteoarthritis (OA) and to determine which is the best view as a research tool in epidemiological studies of knee OA requiring explicit diagnostic criteria to classify the disease in the general population. METHODS: A population based study of 252 unrelated, normal individuals (504 knees) was performed. Lateral and skyline radiographs from each individual were graded for joint space narrowing and osteophytes using a standard atlas. Reproducibility was assessed by two observers on 50 knees. Radiographic features were assessed on their ability to predict knee pain. RESULTS: The skyline views performed better than the lateral views in the assessment of patellofemoral joint OA. The reproducibility for osteophytes was high (kappa > 0.8) and that for joint space narrowing moderate (kappa > 0.6) for both lateral and skyline views. Although the specificity for detecting knee pain was similar in both views, the sensitivity of skyline views in the assessment of knee pain was greater (52.8% versus 30%). The odds ratio for skyline osteophytes as a predictor of knee pain was 7.66 (95% confidence interval (CI) 3.68 to 15.90); that for osteophytes seen on lateral view was 1.83 (95% CI 0.96 to 3.49). Narrowing on both views was a poor predictor of pain. There was frequent disagreement between the lateral and skyline views for detecting osteophytes. CONCLUSION: In a community based study, skyline views performed better than lateral views in terms of reproducibility and for identifying symptomatic patellofemoral joint OA. Skyline radiographs should be the preferred method for examining the patellofemoral joint in such studies. PMID- 8712865 TI - Calcification of the transverse ligament of the atlas in chondrocalcinosis: computed tomography study. AB - OBJECTIVE: To seek an association between articular chondrocalcinosis (AC) and calcification of the transverse ligament of the atlas (TLA), and to evaluate the frequency and the main computed tomography appearances of such calcification. METHODS: Axial computed tomography slices of the cervico-occipital hinge were performed routinely in 21 patients with AC (three men, 18 women; mean age 79 years, range 67-87) and compared with those from a control group of 21 age and gender matched patients without AC. RESULTS: Calcification of the TLA was present in 14 of the 21 patients (66%) in the AC group and in none of the 21 patients (0%) in the control group (chi 2 test: p < 0.001). Calcification was localised behind the odontoid process, inserted upon the osseous tubercles of the lateral masses of C1, and had a curvilinear profile; it varied in height (1.5 to 9 mm) and appearance (thin = < 1 mm; thick = > 1 mm) and formed either a single or a double band. CONCLUSION: This study has demonstrated a relationship between AC and calcification of the TLA. Although such calcification often remains asymptomatic (nine of 14 patients in our study), it may be associated with attacks of acute neck pain with segmentary stiffness, fever, and an increased erythrocyte sedimentation rate, sometimes revealing AC. PMID- 8712866 TI - Prediction of fractures in perimenopausal women: a comparison of dual energy x ray absorptiometry and broadband ultrasound attenuation. AB - OBJECTIVE: To consider whether bone mineral density (BMD) measurements can predict traumatic fractures occurring in perimenopausal women. METHODS: One thousand perimenopausal women called up for screening underwent both dual energy x ray absorptiometry (DXA) of the spine and hip, and broadband ultrasound attenuation (BUA) of the heel. Two years later, they were sent a questionnaire to discover those who had since had a fracture, and compare them with those who had not. RESULTS: About 2% of the women had sustained a fracture in the two years since attendance for screening. Fractures in this age group can be predicted weakly, but significantly, by bone mass measurements using DXA and BUA (odds ratios from 1.4 to 2.1). The lumbar spine appeared to be one of the best predictive sites (odds ratio for 1 SD reduction in BMD 2.1 (95% confidence interval 1.2 to 3.8)), but no significant differences were found between the areas under the curve in receiver operator characteristic (ROC) analysis. CONCLUSION: In this preliminary study it appeared that bone mass measurements are predictive of perimenopausal traumatic fractures in addition to postmenopausal fractures related to osteoporosis. DXA of the lumbar spine did not perform significantly better than BUA. The number of fractures occurring was low, however, and further long term follow up is required to confirm the finding. PMID- 8712867 TI - Changes in bone mass during low dose corticosteroid treatment in patients with polymyalgia rheumatica: a double blind, prospective comparison between prednisolone and deflazacort. AB - OBJECTIVE: To compare the long term effects of low dosage prednisolone or deflazacort treatments on bone mass in patients with polymyalgia rheumatica. METHODS: Thirty patients with polymyalgia rheumatica were allocated on a random double blind basis to receive treatment with prednisolone or deflazacort. Bone mineral content (BMC) was measured in the lumbar spine and in the distal forearm before treatment and three, six, and 12 months after treatment. RESULTS: At three months the decrease in lumbar BMC and bone mineral density (BMD) was significantly greater in the deflazacort group than in the prednisolone group (p < 0.05), but at six and 12 months there was no difference between the two groups. In all patients after one year there was a significant loss of BMC: a 6.4% loss in lumbar BMC and a 1.8% loss in distal forearm BMC. Loss in lumbar BMC after six months was correlated to the cumulative dose of corticosteroid (r = 0.4; p < 0.05) and was significantly greater in the group of patients who had persisting symptoms of polymyalgia at six weeks, three months, or both, after treatment started (p = 0.05). CONCLUSION: This low dose study failed to reveal any calcium sparing properties of deflazacort compared with prednisolone. Possible explanations for this finding are discussed. PMID- 8712868 TI - Increased type I collagen degradation is associated with a need for total joint replacement surgery in rheumatoid arthritis. PMID- 8712869 TI - Polymyalgic presentation of enterovirus infection: a cause of diagnostic confusion. PMID- 8712870 TI - Tenidap: not just another NSAID? PMID- 8712871 TI - The language of rheumatology. II: classification and grouping. PMID- 8712872 TI - Occurrence of rheumatoid arthritis is not increased in the first degree relatives of a population based inception cohort of inflammatory polyarthritis. AB - OBJECTIVE: To determine the risk of rheumatoid arthritis (RA) in first degree relatives of a true population based sample of probands with inflammatory polyarthritis. METHODS: In a case-control study, a two stage screening procedure was used to ascertain the prevalence of RA in 518 first degree relatives of 207 Norfolk Arthritis Register cases registered in 1990 and 414 first degree relatives of 180 local controls. An initial joint symptom and medical history questionnaire was followed by a physical examination, and serological and radiological evaluation of those with symptoms. RESULTS: The prevalence of RA in the first degree relatives of all the Norfolk Arthritis Register cases was 7.7/1000, compared with 4.8/1000 in the first degree relatives of the controls, with a risk ratio of 1.6 (95% confidence interval 0.3 to 8.7). This very modest increase was also seen when the analysis was restricted to the first degree relatives of Norfolk Arthritis Register cases who satisfied the American Rheumatism Association criteria for RA: prevalence rate 7.2/1000. CONCLUSION: There was no evidence of an important increased familial risk of RA in this community based sample. These data are compatible with others from immunogenetic studies showing only weak HLA associations with community ascertained RA. PMID- 8712873 TI - Oral contraception, parity, breast feeding, and severity of rheumatoid arthritis. AB - OBJECTIVE: To investigate the influence of breast feeding, use of the oral contraceptive pill (OCP), and parity on rheumatoid arthritis (RA). METHODS: One hundred and seventy six women with RA were compared with 145 control subjects; all had at least one child. RA patients were classified as having severe (n = 82) or mild disease (n = 89) according to clinical joint evaluation, radiological score, biological inflammation, and the presence of HLA-DR1 or -DR4 alleles. RESULTS: The mean age of RA patients was 58 years, and the mean age at the time of diagnosis of RA was 46 years. The mean time between onset of RA and the first birth was 23.6 (SD 3.8) years. The OCP user rates were 33% in the RA group and 47.6% in the control group (p < 0.02). OCP use was related to the mother's year of birth. The relative risk for developing RA was 0.598 (95% confidence interval (CI) 0.33 to 1.1) in women who had used OCP for more than five years compared with those who had never used OCPs. In contrast, the age at which the first pregnancy occurred, the number of children breast fed, and the duration of breast feeding were comparable in RA patients and healthy subjects. Among the RA patients, parity, duration of breast feeding, and the number of breast fed children were significantly increased in those with severe disease. Having more than three children increased the risk of developing severe disease 4.8-fold when adjusted for age and OCP use. Forty six percent of women with severe RA had a history of breast feeding duration greater than six months before disease onset, compared with 26% of patients with mild disease (p < 0.008). Having more than three breast fed children increased the risk of poor disease prognosis 3.7-fold. In contrast, OCP use had a protective role in the course of RA (44% of RA patients with mild disease were OCP users, compared with 21.7% of those with severe RA; p < 0.001). Among those using OCP for more than five years, the relative risk of developing severe disease was 0.1 (95% CI 0.01 to 0.6), after adjustment for age, parity, and breast feeding. CONCLUSION: Our results suggest that parity, and to a lesser extent breast feeding, before RA onset worsened RA prognosis, whereas OCP use had a protective role. Prolactin and oestrogen may have a role in these effects. PMID- 8712874 TI - Cervical spine surgery in patients with rheumatoid arthritis: an appraisal. AB - OBJECTIVES: To review the outcome of surgery undertaken to stabilise the neck in patients with rheumatoid arthritis performed over a five year period, to compare the results with those of previous reports, and to identify factors that may predict surgical outcome. METHODS: Outcome was assessed at time of discharge from hospital after surgery by review of patients' notes, and at follow up by patient interview, clinical examination, anonymous questionnaire, and cervical spine radiograph. The Ranawat classification of neurological impairment and Steinbrocker functional classification were used. RESULTS: Thirty nine patients underwent 44 procedures; 28 patients were available for review after a mean period of 29.8 months (range 12-65 months). Fourteen patients had preoperative neurological impairment and were available for follow up; 13 returned the questionnaire. Four (29%) had improved Ranawat class, nine were unchanged, and one had deteriorated. Nine (69%) reported a subjective improvement in neurological symptoms by questionnaire, even though the Ranawat class was unchanged in five. Twenty five of the patients reviewed had pain before operation; 21 returned the questionnaire. Pain relief was reported by direct questioning and questionnaire in 76% and 67% of patients, respectively. Overall, 67% felt that surgery had been successful. Surgery was more successful in producing symptomatic relief in patients with neck or radicular pain than in those with neurological deficit, but did prevent progression of neurological symptoms. CONCLUSIONS: Our results are similar to those from other centres. Overall patient satisfaction with surgery was good. Surgery was more likely to produce symptomatic relief in patients with neck or radicular pain before operation than in those with neurological deficit. The greater subjective improvement in neurological symptoms as judged by questionnaire probably reflects the relative insensitivity of the Ranawat classification in detecting change in neurological status; previous reports of poor outcome for patients with neurological symptoms who undergo surgery may in part be a reflection of the insensitivity of this method of assessment. No clear factors emerged which allowed prediction of those patients at greatest risk of operative mortality. In particular, an increased risk of neurological compromise appeared to confer no additional risk of immediate perioperative death. Our data support the suggestion that early surgery to correct symptomatic atlantoaxial subluxation may prevent progression of instability. PMID- 8712875 TI - Haemopoietic stem and progenitor cells in the treatment of severe autoimmune diseases. PMID- 8712876 TI - Chest pain in patients with rheumatoid arthritis. PMID- 8712877 TI - Diagnostic and prognostic characteristics of the enzyme linked immunosorbent rheumatoid factor assays in rheumatoid arthritis. AB - OBJECTIVE: To determine the diagnostic and prognostic test qualities of the enzyme linked immunosorbent assays (ELISA) for rheumatoid factor isotypes in rheumatoid arthritis (RA), and to compare them with the latex fixation test. METHODS: Rheumatoid factor tests were performed in 1988 consecutive new rheumatology outpatients within two months after their first visit to the outpatient clinic of the Department of Rheumatology of Leiden University hospital. The sensitivity, specificity, accuracy, and predictive values of the tests in discriminating RA from non-rheumatoid arthritis and erosive from non erosive disease after two years of follow up were determined and presented as receiver operating characteristic curves and post-test probability curves. RESULTS: The sensitivity of the ELISA for IgG, IgA, and IgM rheumatoid factor for RA versus all controls at optimal cut off titres was 72%, 44%, and 69%, respectively; the specificity was 52%, 84%, and 86%. For the latex fixation test the sensitivity was 66% and the specificity 91%. The post-test probability of RA, at a clinical prevalence rate of 12%, given a positive test result in the ELISAs for IgG, IgA, and IgM rheumatoid factor and the latex fixation test, was 17%, 27%, 40%, and 49%, respectively; with negative test results the probability was 7%, 8%, 5%, and 5%, respectively. The specificity of all tests in discriminating erosive from non-erosive RA at two years was low: 41%, 44%, 47%, and 58% for the ELISAs for IgG, IgA, and IgM rheumatoid factor and the latex fixation test, respectively. CONCLUSION: The ELISAs for IgG and IgA rheumatoid factor are of no significance in diagnosing RA and in the prediction of erosive disease. The ELISA for IgM rheumatoid factor is a reasonable alternative for the latex fixation test when age and gender are taken in to consideration. The specificity of all rheumatoid factor tests in discriminating erosive from non-erosive RA is low. PMID- 8712878 TI - Course and characteristics of anaemia in patients with rheumatoid arthritis of recent onset. AB - OBJECTIVE: To describe the incidence, cause, and course of anaemia in rheumatoid arthritis (RA). METHODS: Medical records of 225 patients who received a diagnosis of RA between 1990 and 1992 were reviewed longitudinally for mention of anaemia. Anaemia was classified as anaemia of chronic disease if ferritin concentrations reflected adequate body iron stores. Among iron depleted anaemic patients, iron deficiency anaemia was identified using the response to iron supplementation. RESULTS: Anaemia developed in 64% of the patients, mostly within 18 months of follow up, but disappeared again in 54% of those patients. The prevalence of anaemia varied from 39% to 53% throughout follow up. Iron depletion was found in 38% of anaemic patients; 40% of them did not recover from their anaemia after iron supplementation and were classified as having anaemia of chronic disease. Anaemia of chronic disease thus caused 77% and iron deficiency anaemia 23% of observed anaemia. Recovery from anaemia occurred in 42% of the patients with anaemia of chronic disease and in 72% of iron depleted patients after iron supplementation. Anaemic patients, particularly those with anaemia of chronic disease, had a significantly greater number of the American College of Rheumatism criteria for RA, significantly more erosive joint damage, and significantly increased concentrations of serum rheumatoid factor than patients without anaemia. CONCLUSION: Anaemia appeared as a frequent and dynamic manifestation. Recovery and recurrence of anaemia was observed throughout follow up, leading to a longstanding and relatively high prevalence of the condition. Iron deficiency was diagnosed frequently and follow up revealed a considerable overlap with anaemia of chronic disease, making this the most important cause of anaemia in RA. Recovery from anaemia occurred more frequently in iron depleted anaemic patients than in those with anaemia of chronic disease. Anaemic patients, particularly those with anaemia of chronic disease, seemed to have a more serious course of their RA compared with non-anaemic patients. PMID- 8712879 TI - A 10 year follow up of parenteral gold therapy in patients with rheumatoid arthritis. AB - OBJECTIVES: To study the long term tolerance of parenteral gold and subsequent drug treatment in patients with rheumatoid arthritis, including prediction of outcome and 'survival' of sequential treatments. METHODS: A retrospective cohort study of 376 patients was made, including a detailed screening of 237 patients treated in 1989. Reasons for discontinuing treatment were analysed in life table analyses, which were used to compare patients receiving parenteral gold treatment in 1985 and 1989, and two groups of patients receiving disease modifying antirheumatic drugs after parenteral gold treatment. The causes of discontinuation were followed in sequential treatments. RESULTS: The estimated probability of discontinuation of parenteral gold treatment was 29% after six months and 42%, 55%, 74%, and 92% after 1, 2, 5, and 10 years, respectively. Mucocutaneous side effects were the main cause of discontinuation of parenteral gold treatment during the first three years, while the probability of discontinuation because of inefficacy dominated after four years. Side effects also constituted the main cause of discontinuation of treatments given after parenteral gold treatment during the first three years of follow up. No significant differences were found when comparing the termination rates between the first and the second and subsequent treatments after parenteral gold treatment. The main reasons for discontinuing one treatment could not predict the cause of discontinuation of the next treatment. CONCLUSION: Mucocutaneous side effects dominated initially, while inefficacy was the dominating cause of discontinuation of long term parenteral gold treatment. No serious side effects were registered. The cause of discontinuation of one treatment did not predict the cause of discontinuation of the following drug. Drug 'survival' was the same in both treatments after parenteral gold treatment. PMID- 8712880 TI - Osteoarticular manifestations of pustulosis palmaris et plantaris and of psoriasis: two distinct entities. AB - OBJECTIVE: To test the hypothesis that pustulosis palmaris et plantaris and psoriatic arthritis (PsA) are two distinct diseases, and that the associated dermatoses are therefore also distinct diseases. METHODS: We prospectively performed clinical, radiological, biological, and bone scan investigations in 23 outpatients with pustolotic arthritis and 23 outpatients with PsA, matched by gender, age (+/- one year) and duration of arthritis (+/- two years). RESULTS: The anterior chest wall, especially the sternocostoclavicular joints, was more frequently involved in pustulotic arthritis than in PsA, both clinically (82% v 43%; p < 0.001) and radiologically (47% v 17%; p < 0.05). Sternocostoclavicular joints generally presented with erosive lesions in PsA, and with large ossifications in pustulotic arthritis. Peripheral joint involvement was mono- or oligoarticular, affecting proximal joints, in pustulotic arthritis (74% v 21%; p < 0.01), and polyarticular, involving small distal joints, in PsA (60% v 0%; p < 10(-4)), in which condition it was also more often erosive (43% v 8%; p < 0.01). The frequency of sacroiliitis and of spine involvement was similar in pustulotic arthritis and PsA. Biology and bone scan did not help distinguish between the two groups. CONCLUSIONS: Pustulotic arthritis and PsA are clinically and radiologically different, therefore pustulosis palmaris et plantaris and psoriasis are most probably distinct dermatological diseases. PMID- 8712881 TI - Effects of transforming growth factor beta s and basic fibroblast growth factor on articular chondrocytes obtained from immobilised rabbit knees. AB - OBJECTIVE: To clarify the effects of transforming growth factor beta 1 (TGF beta 1), TGF beta 2, and basic fibroblast growth factor (bFGF) on cell proliferation and proteoglycan (PG) synthesis in articular chondrocytes obtained from immobilised rabbit knees. METHODS: The right knees of rabbits were immobilised in full extension for up to 42 days using fiberglass casts. Specimens for histology were stained with safranin O. Chondrocytes were isolated from the weight bearing regions of the femur and tibia of the immobilised knees and cultured with combinations of growth factors. Cell proliferation and PG synthesis were determined by 3H-thymidine and 35S-sulphate incorporations. RESULTS: Histological study revealed loss of metachromasia in the articular cartilage at seven days, fissuring and cell clusters at 28 days, and loss of cartilage layers 42 days after immobilisation. Radioisotope assay of the chondrocytes revealed no remarkable change in DNA synthesis in the presence of either TGF beta 1 or TGF beta 2 alone. bFGF markedly stimulated cell proliferation in specimens obtained 0 to seven days after immobilisation. The combination of either TGF beta 1 or TGF beta 2 with bFGF had a synergistic effect, inducing significant increases in DNA synthesis four, seven, and 14 days after immobilisation. PG synthesis by chondrocytes from immobilised joints was not significantly altered by these agents. CONCLUSION: TGF beta 1 or TGF beta 2 in combination with bFGF exert synergistic effects on cell proliferation in articular chondrocytes obtained from the rabbit knee during the early days after immobilisation by a cast. These results suggest a critical role of cytokine combinations in the development of articular cartilage degeneration after immobilisation. PMID- 8712882 TI - Retinal toxicity in long term hydroxychloroquine treatment. AB - OBJECTIVE: To report clinical experience from patients with rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) who were receiving recommended doses of hydroxychloroquine for more than six years, and were monitored for evidence of hydroxychloroquine related retinopathy every six months. METHODS: A prospective (and continuing) evaluation was made of the potential retinal toxicity of hydroxychloroquine in a cohort of 360 Greek patients followed for RA and SLE, 58 of whom have received long term treatment ( > six years). Fundoscopy, colour vision tests, dark adaptation tests, visual field testing, automated perimetry, and electroretinogram were performed every six months. RESULTS: Among 58 patients receiving hydroxychloroquine for more than six years, two relatively young women (3.5%), one treated for RA and the other treated for SLE, developed characteristic hydroxychloroquine related toxic retinal lesions after cumulative doses of 700 g (6.5 years) and 730 g (8 years) of hydroxychloroquine, respectively. Bilateral visual acuity was 6/6 and 6/7.5, respectively; both patients had normal colour perception. Despite an early diagnosis and cessation of treatment, permanent visual field paracentral scotomata in both patients, and persisting lesions in fluorescein angiography in the patient with SLE, were observed at 4.5 and 3 years of follow up, respectively. No other specific cases of hydroxychloroquine related retinopathy have to date been identified in the remaining 302 patients. CONCLUSION: Cases of irreversible, hydroxychloroquine related retinopathy in patients who did not receive overdoses have not been reported previously. The present observations in two relatively young patients should raise our concern regarding the long term usage of an increasingly popular medication in rheumatology practice. PMID- 8712883 TI - Factors associated with the development of vasculitis in rheumatoid arthritis: results of a case-control study. AB - OBJECTIVE: To investigate those characteristics of patients with rheumatoid arthritis (RA) that are associated with the development of rheumatoid vasculitis (RV). METHODS: Demographic and clinical data of 69 patients who had been diagnosed as having RV were compared with those of 138 contemporaneous control patients with RA who were not suspected to have vasculitis. Vasculitis was confirmed histologically in 96% of the subjects with RV. RESULTS: Variables associated with the development of RV were: 1) male gender, presence of increased serum concentrations of rheumatoid factor, joint erosions, subcutaneous nodules, number of disease modifying antirheumatic drugs previously prescribed, treatment (ever) with D-penicillamine or azathioprine; 2) presence of nail fold lesions and any other extrarticular feature one year before the time of diagnosis of RV; 3) treatment with corticosteroids at the time of diagnosis of RV. CONCLUSIONS: The development of RV is associated with male gender, extra-articular features, and a severe course of RA as indicated by the presence of joint destruction and need for intensive treatment with antirheumatic drugs. The strongest association was found with the presence of increased concentrations of rheumatoid factor. PMID- 8712884 TI - High prevalence of arthropathy in HTLV-I carriers on a Japanese island. AB - OBJECTIVE: To determine the aetiological relationship between human T cell leukaemia virus type-I (HTLV-I) and arthritis, by performing an epidemiological study of the inhabitants of Tsushima, an island northwest of the main island of Kyushu, Japan, which is an endemic area of HTLV-I. METHODS: A total of 7087 people underwent an annual health check, and those with arthropathy had further physical and radiological examinations by rheumatologists. The presence of HTLV-I antibody was determined by the particle agglutinin method, and integration of the proviral DNA in peripheral lymphocytes was confirmed by polymerase chain reaction. RESULTS: HTLV-I was positive in 26.1% of inhabitants; the incidence increased to 37.0% in patients with symptoms of polyarthritis. The prevalence of rheumatoid arthritis (RA) was significantly different between HTLV-I carriers (0.56%) and non-carriers (0.31%). The stage of bone destruction in HTLV-I carriers with articular symptoms was milder than that in RA patients without HTLV I. The relative risk of HTLV-I infection for polyarthritis was 1.66 (p < 0.05). CONCLUSION: This is the first epidemiological report clarifying the association between HTLV-I and polyarthritis. Our results suggest that this viral infection has a relationship to RA. PMID- 8712885 TI - Lateral bone density measurements in osteoarthritis of the lumbar spine. AB - OBJECTIVE: To investigate whether spinal osteoarthritis (OA) is responsible for the common finding that lumbar spine bone mineral density (BMD) is greater when measured in the anteroposterior plane than when measured in the lateral plane. METHODS: We studied lateral spine radiographs from 63 women who attended a hospital outpatient department for bone density measurement and who also underwent lumbar spine radiography. Osteoarthritis was assessed using both the Kellgren and Lawrence scale and a scoring system for osteophytosis. Bone density was measured in the anteroposterior and lateral planes using a Hologic QDR-2000 instrument. RESULTS: The mean anteroposterior BMD (0.92 g/cm2) was significantly greater than the lateral BMD (0.59 g/cm2) (p < 0.01), and the difference between antero-posterior and lateral measurements was significantly associated with both increasing Kellgren and Lawrence score and osteophyte score, even after adjustment for age. CONCLUSION: These data suggest that spinal OA is a major cause of the difference between anteroposterior and lateral BMD and that lateral BMD may provide a more accurate representation of true vertebral body bone density in patients with OA of the lumbar spine. PMID- 8712886 TI - Intra-articular guanethidine injection for resistant shoulder pain: a preliminary double blind study of a novel approach. AB - OBJECTIVE: To study the effect of intraarticular (IA) sympathetic blockade for the relief of resistant shoulder pain. METHODS: Eighteen patients with shoulder pain resistant to conventional treatment were allocated randomly to two groups, to receive either IA guanethidine 20 mg or IA saline. They were assessed for pain, and range of active movements, before injection and at one, four, and eight weeks after injection. RESULTS: There were no significant differences between groups, but the group receiving guanethidine showed greater improvement in pain relief at all three follow up visits compared with those receiving placebo (9% v 7% at one week; 15% v 6% at four weeks and 36% v 16% at eight weeks). The improvement reached significance (p < 0.05) at the eight week visit compared with baseline. The range of movement was not significantly improved in either group. CONCLUSION: The results suggest that IA guanethidine produced measurable improvement in resistant shoulder pain and that further studies of this novel approach are indicated. PMID- 8712887 TI - Salivary eicosanoid concentration in patients with Sjogren's syndrome. AB - OBJECTIVE: To investigate eicosanoid concentrations in the saliva of patients with primary Sjogren's syndrome (SS). METHODS: Whole mixed saliva of 36 subjects was assayed for eicosanoid concentrations using a radioimmunoassay. Patients with primary SS having positive lip biopsy served as the study group; their results were compared with data from patients with dry mouth and negative lip biopsy (dry mouth group), and with a group of normal healthy controls. RESULTS: Concentrations of thromboxane B2 were significantly (p < 0.01) increased in 18 patients with primary SS compared with 10 patients with dry mouth and eight healthy normal controls (1.95 (SD 0.51) ng/ml saliva compared with 0.52 (0.1) ng/ml and 0.3 (0.1) ng/ml, respectively). Similarly, prostaglandin E2 concentrations were also significantly increased (p < 0.01) in 11 patients with primary SS compared with five patients with dry mouth and eight normal controls (3.75 (0.82) ng/ml saliva compared with 0.32 (0.1) ng/ml and 0.41 (0.1) ng/ml, respectively). CONCLUSION: Salivary concentrations of eicosanoids are significantly increased in patients with primary SS, and this may prove helpful in the diagnosis of this disease. PMID- 8712888 TI - Antimyeloperoxidase antibodies in individuals with occupational exposure to silica. AB - OBJECTIVE: To determine the prevalence of autoantibodies to myeloperoxidase (MPO) in a series of patients exposed to silica. METHODS: The study included 52 patients with occupational exposure to silica (mean exposure time seven years) and a control group comprising seven patients with progressive systemic sclerosis (PSS), six patients with systemic lupus erythematosus (SLE), and 15 healthy individuals. Antibodies to MPO were detected using commercial enzyme linked immunosorbent assay (ELISA) plates coated with MPO. Indirect immunoflurescence studies for antineutrophil cytoplasmic antibodies were performed using ethanol and formol fixed neutrophils. Clinical and biological data of individuals exposed to silica were recorded (published previously). RESULTS: Antibodies to MPO were detected in 14 individuals exposed to silica (27%). There was a statistically significant difference in anti-MPO ELISA units between the healthy subjects and patients (SLE, PSS, silica exposed individuals) (p < 0.01), but no difference between the different disease groups. CONCLUSIONS: Individuals chronically exposed to silica, whether or not they have a connective tissue disease, have levels of antibodies to MPO (as detected by ELISA) that are greater than those found in the normal population, but similar to those in patients with systemic diseases not induced by silica (SLE/PSS). PMID- 8712890 TI - Infectious disease pathology and emerging infections: are we prepared? PMID- 8712889 TI - Gastrointestinal involvement in Behcet's syndrome: a controlled study. AB - OBJECTIVE: To make a retrospective and prospective analysis of the frequency of symptomatic inflammatory bowel disease in patients with Behcet's syndrome (BS). METHODS: The medical records of the first 1000 patients with BS were reviewed retrospectively for past or present history of diarrhoea. The past and present history of diarrhoea was also elicited prospectively among 147 consecutive patients with BS and 78 diseased controls (42 with rheumatoid arthritis, 17 with systemic lupus erythematosus, seven with seronegative spondylarthropathy, and 12 with miscellaneous rheumatic diseases). Inflammatory mucosal changes were sought in rectal biopsy specimens from 75 patients with BS, 47 diseased controls (29 with nephrotic syndrome, eight with rheumatoid arthritis, six with familial Mediterranean fever, and four with ankylosing spondylitis), and 14 patients with ulcerative colitis. RESULTS: In chart review there were only seven Behcet's patients with diarrhoea; none of them had inflammatory bowel disease. In the prospective survey there were no significant differences between the BS and control groups in the past and present history of diarrhoea. There were no significant differences in the rectal mucosal histology between patients with BS and controls, while patients with ulcerative colitis showed pronounced differences. CONCLUSION: Symptomatic inflammatory bowel disease is not common in BS patients from Turkey. PMID- 8712891 TI - Emerging infectious agents and the forensic pathologist: the New Mexico model. PMID- 8712892 TI - The Army Medical Museum/Armed Forces Institute of Pathology and Emerging Infections: from camp fevers and diarrhea during the American Civil War in the 1860's to global molecular epidemiology and pathology in the 1990s. PMID- 8712893 TI - Retrospective diagnosis of hantavirus pulmonary syndrome, 1978-1993: implications for emerging infectious diseases. AB - OBJECTIVE: To investigate the occurrence of unrecognized cases of hantavirus pulmonary syndrome preceding the detection of the 1993 outbreak in the southwestern United States and the initial description of the syndrome. DESIGN: Retrospective clinicopathologic and immunohistologic study. PATIENTS: Eighty-two patients who died prior to April 1993 with histologically unexplained noncardiogenic pulmonary edema. METHODS: Clinicopathologic review and immunohistochemical evaluation of autopsy tissues for evidence of hantaviral infection. RESULTS: Twelve retrospective fatal cases of hantavirus pulmonary syndrome were identified through clinicopathologic review and immunohistochemical testing of tissues. Patients' ages ranged from 16 to 49 years. The earliest identified case occurred in 1978, 15 years prior to the outbreak of hantavirus pulmonary syndrome in the southwestern United States. Immunohistochemical testing showed widespread deposition of hantaviral antigens, primarily within endothelial cells, similar to the pattern observed with current hantavirus pulmonary syndrome cases. CONCLUSIONS: Although hantavirus pulmonary syndrome was first recognized in 1993, the findings from this study document the earlier existence of this disease. These findings underscore the need for systematic archiving and analysis of clinical information and specimens from patients with diseases of unknown etiology to facilitate the study of new clinical entities and their associated etiologic agents. PMID- 8712894 TI - Lethal experimental infection of rhesus monkeys with Ebola-Zaire (Mayinga) virus by the oral and conjunctival route of exposure. AB - OBJECTIVE: The source of infection or mode of transmission of Ebola virus to human index cases of Ebola fever has not been established. Field observations in outbreaks of Ebola fever indicate that secondary transmission of Ebola virus is linked to improper needle hygiene, direct contact with infected tissue or fluid samples, and close contact with infected patients. While it is presumed that the virus infects through either breaks in the skin or contact with mucous membranes, the only two routes of exposure that have been experimentally validated are parenteral inoculation and aerosol inhalation. Epidemiologic evidence suggests that aerosol exposure is not an important means of virus transmission in natural outbreaks of human Ebola fever; this study was designed to verify that Ebola virus could be effectively transmitted by oral or conjunctival exposure in nonhuman primates. MATERIALS AND METHODS: Adult rhesus monkeys (Macaca mulatta) were exposed to Ebola-Zaire (Mayinga) virus orally (N=4), conjunctivally (N=4), or by intramuscular inoculation (N=1, virus-positive control). RESULTS: Four of four monkeys exposed by the conjunctival route, three of four monkeys exposed by the oral route, and the intramuscularly inoculated positive control monkey (one of one) were successfully infected with Ebola-Zaire (Mayinga). Seven monkeys died of Ebola fever between days 7 and 8 postexposure. One monkey was given aggressive supportive therapy and a platelet transfusion; it lived until day 12 postexposure. CONCLUSIONS: Findings in this study experimentally confirm that Ebola virus can be effectively transmitted via the oral or conjunctival route of exposure in nonhuman primates. PMID- 8712895 TI - Pathology of experimental pneumonic plague produced by fraction 1-positive and fraction 1-negative Yersinia pestis in African green monkeys (Cercopithecus aethiops). AB - OBJECTIVE: The protein capsule of Yersinia pestis, known as Fraction 1 or F1, is a protective immunogen and is an assumed, but not proven, virulence factor. Our objectives were to determine if inhaled F1-negative and/or F1-positive strains of Y pestis were virulent in the African green monkey and, if so, to differentiate F1-negative from F1-positive monkeys. Because F1-negative strains have been isolated from natural sources and have caused experimental fatal disease, we felt that this information was crucial to the development of future vaccines and diagnostic tests. MATERIALS AND METHODS: Adult African green monkeys were exposed by aerosol to F1-positive (CO92, n=15) or F1-negative (CO92-C12, n=6; Java-9, n=2) Y pestis strains. RESULTS: All monkeys died 4 to 10 days postexposure and had lesions consistent with primary pneumonic plague. Antibodies to F1 antigen and other Y pestis antigens allowed us to differentiate F1-positive from F1 negative Y pestis strains in fixed tissues. CONCLUSIONS: In this study, F1 antigen was not a required virulence factor. Therefore, there may be a need for vaccines and diagnostic assays that are not solely based on the F1 antigen. PMID- 8712896 TI - Venezuelan equine encephalitis in BALB/c mice: kinetic analysis of central nervous system infection following aerosol or subcutaneous inoculation. AB - OBJECTIVE: To investigate the routes of entry of Venezuelan equine encephalitis (VEE) virus into the brain, we infected BALB/c mice with a virulent strain (V3000) by aerosol or subcutaneous inoculation. METHODS: Immunohistochemistry and in situ hybridization methods were used to detect VEE virus in tissues taken at daily intervals postinfection. RESULTS: In both groups, virus in the brain first appeared in olfactory regions. Aerosol exposure caused early massive infection of olfactory epithelium, which developed into bilaterally symmetrical infection of the olfactory nerves, olfactory bulbs, and lateral olfactory tracts by day 2 postinfection. After subcutaneous inoculation, VEE in the brain also appeared first in olfactory regions, but was not detected until day 3 postinfection. By day 4 postinfection, VEE viral infection had spread throughout the brain in both groups. Vascular endothelium and the choroid plexus remained uninfected during the entire study. CONCLUSIONS: Our findings suggest that VEE virus, whether given by aerosol or subcutaneously, first enters the brain through the olfactory tract. PMID- 8712897 TI - Pathology of microsporidiosis: emerging parasitic infections in patients with acquired immunodeficiency syndrome. AB - OBJECTIVE: Microsporidiosis is a group of rapidly emerging protozoan infections that have thus far been reported predominantly from severely immunosuppressed persons with the acquired immunodeficiency syndrome (AIDS). The four genera that have been identified in AIDS patients (Enterocytozoon, Encephalitozoon, Septata, and Pleistophora) are an increasingly common source of both localized and disseminated infections. However, the clinical and pathologic features of these agents are being described with such rapidity that many pathologists are unaware of the histologic, immunologic, and molecular methods for diagnosing these infections. This article summarizes the clinical and morphologic spectrum of the microsporidian species that infect patients with AIDS. Additionally, the role of ultrastructural, immunologic, tissue culture, and molecular techniques for the diagnosis of microsporidian infections are discussed. DATA SOURCES: Clinical and pathologic findings were obtained from patients with AIDS who were evaluated for microsporidian infections at the Grady Memorial Hospital in Atlanta. Selected laboratory studies were performed at the Division of Parasitic Diseases of the Centers for Disease Control and Prevention and at the Department of Physiology at Morehouse University. Additionally, some cases were sent for consultation to the Infectious Disease Pathology service at Emory University. These data were combined with the published studies of microsporidian infection from the medical literature. DATA SYNTHESIS: The pathologic appearance of microsporidian infections in each major organ system (ocular, respiratory, genitourinary, gastrointestinal) is illustrated using routine and special histochemistry and immunofluorescence. The differential diagnostic features of the four genera of microsporidia infecting AIDS patients are illustrated using transmission and scanning electron micrographs from biopsy, autopsy, and tissue culture materials. Cytologic evaluation of body tissues is emphasized as a sensitive method for microsporidian diagnosis. CONCLUSIONS: Microsporidian infections can be expected to remain an increasingly important cause of morbidity and mortality in patients with AIDS. It is important that pathologists and microbiologists become acquainted with the clinicopathologic spectrum of these emerging protozoal infections, ensuring timely diagnosis and subsequent treatment. PMID- 8712899 TI - Autopsy verification of Encephalitozoon intestinalis (microsporidiosis) eradication following albendazole therapy. AB - Microsporidian infections are increasingly recognized as an important cause of morbidity for persons infected with the human immunodeficiency virus. Encephalitozoon (formerly Septata) intestinalis is a recently described microsporidian that causes intestinal and disseminated infections in severely immunocompromised patients with acquired immunodeficiency syndrome. Several studies suggest that albendazole is an effective therapy for E intestinalis infection. However, relapses of symptoms and reappearance of microsporidian spores in diagnostic specimens have been reported following treatment in some cases. Because these results are based on examination of feces or cytologic specimens with an inherent sampling bias, it would be ideal to have autopsy data on the complete tissue evaluation of major organ systems of patients who had antemortem E intestinalis infection treated with albendazole. This report describes an acquired immunodeficiency syndrome patient with diarrhea and wasting syndrome associated with E intestinalis infection. Treatment with albendazole produced relief of his clinical symptoms and eliminated microsporidian spores in his feces. Following his death from other causes, an autopsy was performed. Comprehensive microscopic examination of all major organs revealed no evidence of residual microsporidian infection, suggesting parasitologic cure of E intestinalis with albendazole. The postmortem finding of complete clearance of microsporidia from body tissues is significant for future albendazole treatment of patients infected with E intestinalis and provides strong support for the value of the autopsy in evaluating the therapeutic efficacy of antimicrobials in emerging infections. PMID- 8712900 TI - Granulomatous appendicitis in acute myeloblastic leukemia: expanding the clinicopathologic spectrum of invasive candidiasis. AB - The incidence of systemic candidiasis appears to be on the rise, and unusual or hitherto undocumented clinicopathologic features of invasive candidiasis continue to be described. This report adds further to this spectrum by describing granulomatous appendicitis due to invasive candidiasis. The patient was a 23-year old woman with acute promyelocytic leukemia. Her other risk factors included the use of aggressive chemotherapy, immunosuppressives, broadspectrum antibiotics, and invasive instrumentation. In view of the acquired immunodeficiency syndrome epidemic, as well as the improved survival of patients with prolonged neutropenia owing to better intensive care units, there is reason to believe that other as yet undocumented clinicopathologic manifestations of invasive candidiasis or other opportunistic infections may come to light. PMID- 8712898 TI - Visceral leishmaniasis emerging as an important opportunistic infection in HIV infected persons living in areas nonendemic for Leishmania donovani. AB - BACKGROUND: Visceral leishmaniasis is an important infection in patients infected with human immunodeficiency virus and living in areas endemic for Leishmania sp. Leishmaniasis, however, is rarely suspected in patients residing in nonendemic countries. METHODS: Retrospective case analysis of 15 patients with human immunodeficiency virus infection and leishmaniasis treated at seven German clinics. The clinicopathological features and the diagnostic role of biopsy and/or cytology as compared to serology were evaluated. RESULTS: All patients were severely immunocompromised. One patient was first diagnosed at autopsy. One patient with mucocutaneous disease was diagnosed by nasal biopsy. All others had amastigotes detected in bone marrow (13/13), liver (3/3), and gastrointestinal mucosa (4/4). Serology was positive in only 6 or 13. CONCLUSION: Visceral leishmaniasis is an important opportunistic infection in patients with acquired immunodeficiency syndrome and it must be ruled out in every patient with fever and/or pancytopenia and an appropriate travel history. Because serological diagnosis is often insufficient, pathologists must be aware of the association between human immunodeficiency virus infection and leishmaniasis. Diagnosis depends on detection of the parasite in submitted specimens. PMID- 8712901 TI - The use and abuse of routine stool microbiology: a College of American Pathologists Q-probes study of 601 institutions. AB - OBJECTIVE: To examine the efficiency with which physicians use routine stool microbiology tests. DESIGN: Questionnaire and structured review of 100 consecutive stool bacteriology and parasitology examinations at each participating institution. SETTING: Six hundred one institutions enrolled in the College of American Pathologists Q-probes Program. RESULTS: Of 59500 bacteriology specimens, 3808 (6.4%) contained a pathogen. The vast majority (99%) of bacterial pathogens were detected in either the first or second specimen submitted. Almost 40% of inpatient specimens were collected after the third day of hospitalization, but only 0.6% of these specimens were positive for enteric pathogens that had not been previously recovered. More than half of the laboratories reported having no limits on the number of bacteriology specimens per patient that could be submitted for testing, and fewer than 8% of laboratories rejected specimens from inpatients after a certain number of days in the hospital. The frequency with which laboratories performed tests for Clostridium difficile varied widely. Of 58500 parasitology specimens, 1463 (2.5%) contained a pathogen; 97.6% of pathogens were detected by the second stool specimen, and 99.8% were detected by the third specimen. Only 0.7% of specimens from inpatients hospitalized more than 4 days contained a new pathogen. CONCLUSIONS: We recommend that no more than two bacteriology specimens and no more that two or three parasitology specimens be processed per patient without consultation. Standard stool examination for a bacterial pathogens has a low yield and should not be performed after 3 days of hospitalization. Likewise, parasitology examinations should not be performed after 4 days of hospitalization. PMID- 8712902 TI - Retrospective review of colorectal cancer in ulcerative colitis at a tertiary center. AB - OBJECTIVE: To review patient characteristics, surgical indications, operative procedures, and survival of patients with ulcerative colitis who develop colorectal cancer. DESIGN: Retrospective medical record review. SETTING: Tertiary referral center. PATIENTS: Of 493 patients who underwent surgery for ulcerative colitis between 1978 and 1994, 25 patients had colorectal cancer. INTERVENTION: All patients underwent surgical exploration and either a biopsy, segmental resection, total abdominal colectomy, or restorative proctocolectomy was performed. MAIN OUTCOME MEASURE: Duration of postoperative disease-free survival. RESULTS: The average duration of illness prior to surgery was 18.5 years (range, 0.25-40 years). Surgical indications were intractability for 3 patients (12%); dysplasia as revealed by colonoscopic biopsy for 8 patients (32%); and preoperatively diagnosed cancer for 14 patients (56%). Three patients (12%) underwent exploratory laparotomy, 3 (12%) underwent right hemicolectomy, 5 (20%) underwent total proctocolectomy with ileostomy, 3 (12%) underwent total proctocolectomy with continent ileostomy, and 12 (48%) underwent restorative proctocolectomy. Pathological stage was carcinoma in situ for 3 patients (12%), stage I for 7 patients (28%), stage II for 4 patients (16%), stage III for 6 patients (25%), and stage IV for 5 patients (20%). Five-year survival was 100% (3/3) for patients with carcinoma in situ, 100% (4/4) for those with stage I disease, 50% (2/4) for those with stage II, 25% (1/4) for those with stage III, and 0% (0/5) for those with stage IV. Of the 12 patients treated with a restorative proctocolectomy, 6 are 5-year survivors, and 4 are alive at 24, 36, 38, and 48 months. CONCLUSIONS: Twelve percent of the patients in this series had no preoperative evidence of colorectal malignant neoplasms but had invasive cancer in the resected specimen. Therefore, duration of disease alone may be an indication for surgery. A restorative proctocolectomy is a satisfactory procedure in selected patients with malignant neoplasms. PMID- 8712903 TI - Assessment of resectability of pancreatic head and periampullary tumors by color flow Doppler sonography. AB - OBJECTIVE: To examine the sensitivity of color flow Doppler ultrasonography in assessing resectability of pancreatic head and periampullary tumors. DESIGN: Validation cohort study. SETTING: Tertiary care public hospital. PATIENTS: Thirty seven patients with pancreatic head or periampullary cancer were studied by color flow Doppler examination of the relevant blood vessels. MAIN OUTCOME MEASURE: A pancreatic Doppler score (PDS) was defined as the closest circumferential contact of the tumor to the superior mesenteric vein, superior mesenteric artery, or portal vein. A PDS of 1 indicated no contact (n = 9); PDS 2, less than 50% contact (n = 10); PDS 3, 50% to 99% contact (n = 7); and PDS 4, encasement (n = 11). The PDS was compared with operative and histologic resection margins. RESULTS: The lack of vascular invasion was confirmed operatively in 7 of 7 patients with a PDS of 1, and 6 patients who underwent resection had clear histologic margins. Nine (90%) of 10 patients with a PDS of 2 were confirmed to have no vascular invasion, and 3 (43%) of 7 patients who underwent resection had clear margins. Five (83%) of 6 patients with a PDS of 3 had correct operative findings, and both patients who underwent resection had positive margins. Operative confirmation of encasement was found in all 7 patients with a PDS of 4 who had operative exploration, and none underwent resection. CONCLUSIONS: Color flow Doppler sonography and PDS predicted resectability and the histologic margin status (positive predictive value, 97%). Patients with a PDS of 1 are predicted to have clear histologic margins after resection. Patients with a PDS of 4 have unresectable tumors, and nonoperative palliation should be considered. Patients with a PDS of 2 or 3 have a high likelihood of positive histologic margins after resection and may be candidates for neoadjuvant chemotherapy. PMID- 8712904 TI - The role of neoadjuvant therapy in surgically resectable esophageal cancer. AB - OBJECTIVE: To determine the effect of neoadjuvant therapy (NT) (preoperative chemotherapy, radiation therapy, or both) in surgically resectable esophageal cancer. DESIGN: A retrospective review over a 20-year period. SETTING: A tertiary academic medical center. PARTICIPANTS: All patients undergoing surgical resection for esophageal cancer (N = 316) over this time period. MAIN OUTCOME MEASURES: Perioperative morbidity and mortality, local and distant recurrences, and overall survival. RESULTS: Patients undergoing NT (n = 106) had prognostic factors similar to those treated with surgery alone (n = 210). No increase was noted in surgical morbidity with NT (anastomotic leaks, reoperation rates, complications, or extended hospital stays). Overall survival was not improved by NT (median survival, 14 months) except in the subset of patients (11/83) who responded completely (100% histological necrosis) to preoperative chemotherapy (median survival, 79.2 months; P < .02). Complete response to radiation therapy alone was not associated with improved survival. Partial necrosis of the primary tumor was seen in 13 (15%) of 83 patients but conferred no survival advantage. Complete response to preoperative chemotherapy was associated with squamous cell pathological features and excellent performance status as measured by preanesthesia evaluation. CONCLUSIONS: The addition of NT did not increase perioperative morbidity or mortality. Only the subset of patients who had a complete response to preoperative chemotherapy showed a survival advantage. Excellent performance status and squamous cell pathological features were associated with an increased chance of complete pathological response following preoperative chemotherapy. PMID- 8712905 TI - Safe laparoendoscopic approach to biliary pancreatitis in older patients. AB - OBJECTIVE: To evaluate management strategies for biliary pancreatitis in different age groups. DESIGN: Retrospective review. SETTING: Large private metropolitan teaching hospital. PATIENTS: Patients seen between January 1991 and December 1994 with a diagnosis of biliary pancreatitis (N = 136) divided into 2 groups (group 1, aged < 65 years; group 2, aged > or = 65 years). INTERVENTIONS: Primary treatments included endoscopic retrograde cholangiography (n = 36) alone or with endoscopic sphincterotomy (n = 27); operative procedures, including cholecystectomy by laparoscopic (n = 54) or open (n = 16) approaches; or no definitive therapy (n = 22). Secondary treatments of common bile duct stones included laparoscopic transcystic bile duct exploration (n = 5), open common bile duct exploration (n = 4), or postoperative endoscopic retrograde cholangiography (n = 10). MAIN OUTCOME MEASURES: Success of interventions, incidence and treatment of common bile duct stones, morbidity and mortality rates, frequency of retained stones, and length of hospitalization. RESULTS: Numbers of Ranson criteria were higher for older patients (group 1, 0.83 +/- 0.12 vs group 2, 1.57 +/- 0.11 [mean +/- SEM]; P < .001). Primary endoscopic retrograde cholangiography with or without endoscopic sphincterotomy was performed earlier than operative procedures, with a significantly higher incidence of common bile duct stones (72% vs 19%; P < .001). Number of primary procedures and complication and mortality rates for endoscopic retrograde cholangiography with or without endoscopic sphincterotomy were 36, 8%, and 3%, respectively; for laparoscopic cholecystectomy, 54, 9%, and 2%, respectively; and for open cholecystectomy, 16, 6%, and 19%, respectively. For complication and mortality rates, there were no statistical differences between groups or among treatments. Deferred therapy was used in 30 patients, with 20% readmitted for recurrence of biliary pancreatitis. Length of intensive care unit and total hospital stay were similar for all groups and treatments. CONCLUSIONS: Older patients with biliary pancreatitis may be safely treated with a combined laparoendoscopic approach. Management of common bile duct stones depends on age, with laparoscopic transcystic duct exploration or open common bile duct exploration preferred for younger patients and laparoscopic transcystic duct exploration or postoperative endoscopic sphincterotomy for older ones. Deferred therapy has a substantial relapse rate. PMID- 8712906 TI - Left-upper-quadrant devascularization for 'unshuntable' portal hypertension. AB - BACKGROUND: No simple solution exists for the patient with bleeding due to diffuse splanchnic venous thrombosis (so-called unshuntable portal hypertension). Radical gastroesophageal devascularization or extended esophagogastrectomy has been considered obligatory in this setting. OBJECTIVE: To examine the use of 1 stage, left-upper-quadrant devascularization for unshuntable portal hypertension. DESIGN: A retrospective call-back survey. SETTING: A regional referral center. PATIENTS: Eight consecutive patients with recurrent bleeding from esophagogastric varices due to diffuse splanchnic venous thrombosis. INTERVENTIONS: Splenectomy, staple transection of the esophagus, and proximal gastric devascularization. MAIN OUTCOME MEASURES: Operative complications, recurrent bleeding, survival, and quality of life. RESULTS: No operative deaths occurred, and 7 of 8 patients who were treated for unshuntable portal hypertension and who were followed-up for 1 to 15 years (mean, 4.7 years) are alive. No patient has had a recurrent variceal hemorrhage. A second endoscopy has demonstrated small varices in 4 patients. Early and late complications occurred in 3 and 1 of the patients, respectively. CONCLUSIONS: Left-upper-quadrant devascularization is a technically straightforward, safe, effective, and durable alternative to the Sugiura procedure or to radical esophagogastrectomy in patients with unshuntable portal hypertension. PMID- 8712907 TI - Improved results of liver transplantation in patients with portal vein thrombosis. AB - OBJECTIVE: To analyze the impact of preexisting portal vein thrombosis (PVT) on the operative management and outcome of liver transplantation. DESIGN: Retrospective review of 1423 patients who received transplants over 11 years. SETTING: Tertiary referral center. PATIENTS OR OTHER PARTICIPANTS: Seventy patients who underwent liver transplantation who had preexisting PVT. INTERVENTIONS: Portal vein thromboendovenectomy, vein grafting, or use of portal collateral veins for inflow during liver transplantation. MAIN OUTCOME MEASURES: Postoperative PVT, intraoperative transfusion, retransplantation rate, 30-day and 1-year actuarial survival rates. RESULTS: Operative management consisted of thromboendovenectomy in 61 cases, vein graft to the superior mesenteric vein in 6 cases, and vein graft to other mesenteric veins in 3 cases. The incidence of posttransplant PVT was 3% (n = 2). The mean +/- SD transfusion requirement was 23 +/- 18 U. The 1-year actuarial survival rate was 74% but improved from 66% in the first 35 cases to 82% in the latter 35 cases. CONCLUSIONS: Thromboendovenectomy is the procedure of choice for PVT. Results of liver transplantation in patients with PVT improve significantly with experience gained and are equivalent to results in patients without PVT. PMID- 8712908 TI - Moments in surgical history: Arthur Dean Bevan. PMID- 8712909 TI - Necrotizing soft-tissue infections. AB - OBJECTIVES: To evaluate the causes of necrotizing soft-tissue infections (NSTls) and to determine the outcomes of patients with NSTls. DESIGN: A restrospective survey of the causes and factors associated with the outcomes. SETTING: An urban community hospital serving an indigent population. PATIENTS: A consecutive series of patients with NTSls who were treated between December 11, 1990, and December 28, 1995. INTERVENTIONS: Patients were treated with operative debridement, intravenous antibiotics, and supportive measures. MAIN OUTCOME MEASURES: Patient outcomes, causes, the extent of infection, the health status of the patients, causative organisms, and treatment delays. RESULTS: Forty-five patients with NSTls were identified. Twenty-eight cases (62%) have occurred since January 13, 1994. Parenteral drug abuse, the causative event in 25 cases (56%), accounted for 21 (75%) of the 28 cases identified since January 13, 1994. Skin flora were the primary isolates in 18 (40%) of the cases; 78% of these flora were polymicrobial. Clostridial species were isolated in 8 (18%) of the cases. The overall mortality was 27%. Survivors had less extensive infections and were more stable hemodynamically than nonsurvivors. Patients with necrosis and cellulitis greater than 250 cm2 were less likely to survive than those with less extensive infections. Logistic regression analysis identified the extents of infection, the initial blood pressure, and the initial temperature as independent predictors of outcome in this patient series. CONCLUSIONS: This is 1 of the largest reported series of patients with NSTls in which parenteral drug abuse is a prevalent causative factor. The proportion of NSTls attributable to the injection of illicit substances has increased notably in the past 2 years and has reached epidemic proportions. Survivors of NSTls had less extensive infections and were more often hemodynamically stable than nonsurvivors. Clostridial species were common in patients with NSTls related to parenteral drug abuse, underscoring the need for awareness of the potential for wound botulism in these patients. PMID- 8712910 TI - Inhaled nitric oxide prevents experimental platelet activating factor-induced shock. AB - OBJECTIVE: To investigate whether inhaled nitric oxide (INO) can prevent platelet activating factor (PAF)-induced pulmonary hypertension and shock. DESIGN: Randomized controlled animal trial. SETTING: Laboratory. SUBJECTS: Yorkshire swine. INTERVENTIONS: Animals received general anesthesia and invasive hemodynamic monitoring, then PAF only, 2.5 micrograms/kg intravenously over 45 minutes (PAF group, n = 9) or PAF in addition to INO, 20 ppm (PAF-INO group, n = 6). MAIN OUTCOME: Vascular pressures (mean arterial and mean pulmonary), vascular resistance indexes (systemic and pulmonary), cardiac indexes, and oxygen delivery and oxygen consumption. RESULTS: Mean arterial pressures, cardiac indexes, and oxygen delivery and consumption were significantly higher in the PAF-INO group. Mean pulmonary arterial pressures and systemic and pulmonary vascular resistance indexes were significantly lower in the PAF-INO group. There were 4 deaths (44%) in the PAF group vs none (0%) in the PAF-INO group (P = 10). CONCLUSIONS: The use of INO prevents pulmonary hypertension, circulatory failure, and death during PAF induced shock. PMID- 8712911 TI - Reoperation for persistent or recurrent primary hyperparathyroidism. AB - OBJECTIVE: To analyze the causes and outcomes of reoperation for persistent or recurrent primary hyperparathyroidism. DATA SOURCES: Medical records of 102 patients with persistent or recurrent primary hyperparathyroidism who underwent reoperation by 1 surgeon between 1985 and 1995. STUDY SELECTION: Only patients with persistent or recurrent primary hyperparathyroidism were selected; patients with secondary hyperparathyroidism, parathyroid cancer, familial hyperparathyroidism, and previous thyroid operations were omitted. DATA EXTRACTION: Performed by a single unblinded researcher. DATA SYNTHESIS: Reasons for failed parathyroid operations included tumor in ectopic position (53%), incomplete resection of multiple abnormal glands (37%), adenoma in normal position missed during previous surgery (7%), and regrowth of previously resected tumor (3%). Of the ectopic glands, 28% were paraesophageal, 26% in the mediastinum (nonthymic), 24% intrathymic, 11% intrathyroidal, 9% in the carotid sheath, and 2% in a high cervical position. Eighty-three percent of ectopic glands were accessible via cervical incision. The success rate of reoperations was 95%. One patient (1%) became permanently hypocalcemic after reoperation; 1 patient (1%) suffered permanent unilateral vocal cord paralysis. The sensitivities of preoperative localization studies were as follows: technetium Tc 99m sestamibi scan, 77%; magnetic resonance imaging, 77%; selective venous catheterization for intact parathyroid hormone, 77%; thallium-technetium scan, 68%; ultrasonography, 57%; and computed tomography, 42%. CONCLUSIONS: Repeated parathyroidectomy can be avoided in more than 95% of patients if an experienced surgeon performs bilateral cervical exploration during the initial parathyroid operation. For patients with persistent or recurrent primary hyperparathyroidism, preoperative localization studies and a focused surgical approach can result in a 95% success rate with minimum complications. PMID- 8712912 TI - Laparoscopic adrenalectomy. Comparison of the lateral and posterior approaches. AB - OBJECTIVE: To compare the lateral transabdominal and posterior retroperitoneal laparoscopic methods for performing adrenalectomy. DESIGN: Nonrandomized. SETTING: Hospitals affiliated with the University of California, San Francisco. PATIENTS: Thirty-six patients (15 men and 21 women), aged 5 to 78 years (mean age, 49 years), were treated for the following conditions: aldosteronoma, 18 patients; pheochromocytoma, 4 patients; Cushing syndrome, 6 patients; androgen secreting tumor, 1 patient; nonfunctioning adenoma, 3 patients; adrenal hemorrhage, 1 patient; metastatic neoplasm, 2 patients; and myelolipoma, 1 patient. INTERVENTIONS: Twenty-three lateral and 14 posterior laparoscopic adrenalectomies. MAIN OUTCOME MEASURES: Success rate, operating time, complications, and length of hospital stay. RESULTS: The tumors, which ranged in size from 1 to 13 cm (mean, 4.2 cm; median, 2.5 cm), were all successfully resected laparoscopically. All 8 tumors larger than 6 cm were resected by the lateral approach. One critically ill patient died. No patient required blood transfusions or conversion to laparotomy. Mean operating time was 3.8 hours vs 3.4 hours (median, 3.5 hours vs 3 hours) and mean hospital stay was 2.2 days vs 1.5 days (median, 2 days vs 1 day) for the lateral and posterior approaches, respectively. All patients without concomitant procedures were ready to be discharged within 48 hours. CONCLUSIONS: Both approaches were effective and safe. We prefer the lateral approach for tumors larger than 6 cm and the posterior approach for bilateral tumors. PMID- 8712914 TI - Long-term results after colectomy and ileoanal pull-through procedure in children. AB - OBJECTIVE: To review the clinical experience with colectomy and ileoanal pull through procedure in children from 1 hospital. DESIGN: Since 1977, 116 children 18 years of age or younger underwent colectomy and ileoanal pull-through procedure at University of California-Los Angeles Medical Center. Ninety-four children had ulcerative colitis, 17 had familial polyposis coli, and 5 had Hirschsprung disease. Sixty-two children had a lateral pouch, 47 a J-pouch, and 7 a straight pull-through. A diverting ileostomy was used for 4 months for all patients except 9 with polyposis coli and 2 with Hirschsprung disease. During the same period, an additional 414 patients older than 18 years underwent the ileoanal pull-through procedure. RESULTS: Forty-eight children (41%) developed complications; the most common was pouchitis, which occurred in 18 patients with ulcerative colitis. Forty-two children underwent reoperation; 38 had ulcerative colitis. There were no deaths. Six children (5.2%) (3 with Crohn disease) required a permanent ileostomy. Six straight pull-throughs were converted to J pouches because of stool frequency; 19 patients with lateral pouches underwent pouch reconstruction or spout resection because of stasis. With a mean follow-up of 7.1 years, 107 children (92.2%) were progressing well. CONCLUSIONS: Ulcerative colitis is a more severe disease in children. The ileoanal pull-through procedure is the preferred operation for children with ulcerative colitis, polyposis coli, and selected patients with Hirschsprung disease. The J-pouch is preferred because of simplicity of construction and scarcity of complications. PMID- 8712913 TI - Preresection chemotherapy improves survival for children with Askin tumors. AB - OBJECTIVE: To test whether patients with Askin tumor treated with aggressive neoadjuvant chemotherapy have a better clinical outcome. DESIGN: Retrospective case series. SETTING: Pediatric referral center. PATIENTS: All children diagnosed with malignant small-cell tumors of the chest wall (Askin tumor) and treated from 1975 to September 1987 (phase 1, n = 6) and from September 1987 to the present (phase 2, n = 9). MAIN OUTCOME MEASURES: Survival as a function of extent of disease and response to therapy as measured by tumor volume, survival, and recurrence. RESULTS: All phase 2 patients had significant reduction of tumor volume and improved survival by Kaplan-Meier estimates compared with phase 1 patients. No phase 1 patients are still alive. CONCLUSION: Patients with Askin tumor treated with aggressive preresection chemotherapy have smaller tumors to resect and improved survival. PMID- 8712915 TI - Decreased mortality from technical failure improves results in pediatric liver transplantation. AB - BACKGROUND: Until recently, pediatric liver transplantation was associated with a high rate of technical failure, which contributed substantially to the overall prognosis. OBJECTIVE: To assess the impact of technical failure on outcome in pediatric liver transplantation. DESIGN AND SETTING: We retrospectively analyzed 90 pediatric transplant procedures in a university medical center. PATIENTS: Between February 1988 and December 1995, 80 children ( < 15 years old) received 90 transplants. Fifty-three percent (n = 42) were less than 2 years of age, 45% (n = 36) had cholestatic liver disease, 26% (n = 21) had metabolic errors, and 11% (n = 9) had fulminant hepatitis. INTERVENTION: Patients underwent grafting using previously reported techniques, including cadaveric whole (61% [n = 55]), reduced-size (17% [n = 15]), and living related (22% [n = 20]) liver transplantation. MAIN OUTCOME MEASURES: Patient and graft survival and selected surgical complications. Outcomes were compared before (group 1) and after (group 2) the introduction of living related transplantation in July 1992. RESULTS: In group 1, 32 patients received 36 grafts (4 retransplants [13%]), and in group 2, 48 patients received 54 grafts (6 retransplants [13%]). Six- and 12-month patient survival rates were 78% (n = 25) and 75% (n = 24), respectively, for group 1 and 98% (n = 47) and 94% (n = 45) for group 2. Of the 9 deaths in group 1, 6 occurred early as a consequence of surgical complications, while in group 2, all 5 deaths that occurred were caused by the consequences of immunosuppression (lymphoproliferative disease, n = 2; late infections, n = 3). CONCLUSIONS: These results suggest that mortality caused by surgical complications has been reduced by improvement in management in recent years. Living related grafts have supplemented the graft supply and may be associated with the improved overall results. Despite these advances, children receiving transplants continue to experience the consequences of imperfect immunosuppression. PMID- 8712916 TI - Peroneal bypass is equivalent to inframalleolar bypass for ischemic pedal gangrene. AB - OBJECTIVE: To determine if peroneal bypass is a suitable alternative to inframalleolar bypass in patients with ischemic pedal gangrene. DESIGN: Review of a prospectively acquired vascular registry. SETTING: University practice limited to vascular surgery. PATIENTS: Patients with chronic lower-extremity ischemia and pedal gangrene evaluated between 1985 and 1995 in whom the only options for arterial reconstruction were bypass to the peroneal or an inframalleolar artery. INTERVENTIONS: Peroneal or inframalleolar reverse vein bypass. MAIN OUTCOME MEASURES: Time to healing and lifetable analyses of survival, primary patency, and limb salvage. RESULTS: Eighty-three peroneal and 46 pedal bypasses were performed for ischemic foot gangrene. The groups were equivalent for sex, diabetes mellitus, heart disease, hypertension, renal failure, hypercoagulable states, previous ipsilateral bypass, smoking, and preoperative ankle-brachial indices. Patients with inframalleolar bypass were younger than patients with peroneal bypass (63.9 vs 71.6 years, P = .005) and had higher postoperative ankle brachial indices (1.02 vs 0.91, P = .004). However, 3-year survival rates (69.1% inframalleolar vs 60.0% peroneal, P = .35), limb salvage rates at 2 years (70.3% vs 85.8%, P = .10), and time to wound healing (19.7 vs 21.6 weeks, P = .66) were equivalent. CONCLUSION: Peroneal and inframalleolar bypass for ischemic pedal gangrene have equivalent intermediate-term survival, limb salvage, and wound healing. Surgeons should not feel obliged to perform inframalleolar bypass for pedal gangrene if peroneal bypass is possible. PMID- 8712917 TI - Code of Ethics. Royal Australasian College of Surgeons, September 1993. PMID- 8712918 TI - Mapping of transregulatory domains of pseudorabies virus early protein 0 and identification of its dominant-negative mutant. AB - Pseudorabies virus (PRV) early protein 0 (EP0) is a transactivator containing the RING finger domain. Analysis of transactivating activity of truncated forms of the EP0 molecule consisting of 410 amino acids revealed that amino-terminal region containing the RING finger domain, amino acids 1 to 84, and the region between amino acids 114 to 242 containing acidic amino acid sequences were required for the transactivation. On the other hand, the mutant consisting of amino acids 1 to 113 exhibited a dominant-negative property. PMID- 8712919 TI - Selective cleavage by trypsin of the capsid protein VP1 of type 3 poliovirus results in improved sorting of cell bound virions. AB - A large proportion of host cell-bound virions of poliovirus type 1 strain Mahoney (PV1/M) is known to elute to the culture medium during incubation at 37 degrees C, and only a fraction of the virions remaining cell-associated will successfully uncoat and contribute to the new replication cycle. We found that while the proportion of inoculum type 3 poliovirus strain Saukett (PV3/S) bound to GMK cells was of the same order as that of PV1/M, the bound PV3/S virions uncoated much less efficiently, as judged by velocity sedimentation analysis of virion disintegration. Rather, the majority of the cell-associated PV3/S viruses remained apparently unaffected for several hours within an unidentified intracellular compartment. Incubation of PV3/S with intestinal trypsin is known to result in selective cleavage of the capsid protein VP1 and striking antigenic changes. Trypsin treatment of stock PV3/S preparations did not affect the infectivity titre or modify single-cycle progeny virus yields significantly. However, the fate of the cell-bound inoculum virus was profoundly altered. Trypsin-treated PV3/S virions (PV3/S-Try) attached to GMK cells less tightly than the untreated PV3/S virus or PV1/M, and a relatively larger proportion of the cell-bound virus eluted to the medium during subsequent incubation at 36 degrees C. However, the fraction of virions remaining cell-associated rapidly disintegrated suggesting efficient uncoating. In accordance with these observations, one step growth curves of PV3/S-Try in all cell lines tested showed lowered eclipse phase titres compared to those obtained with the untreated PV3/S inocula. Similar effects were also demonstrated for type 3 poliovirus strain Sabin while trypsin-sensitive strains of the other two serotypes of poliovirus remained unaffected in this sense. The putative biological significance of the altered sorting of cell-bound PV3/S-Try virions is not known. It might be related to the observations that sensitivity of type 3 poliovirus strains to trypsin is conserved in spite of the fact that the target site of trypsin action is flanked by highly variable motives in an immunodominant antigenic site. PMID- 8712920 TI - Genome polymorphism of human adenoviruses of subgenus C. AB - The genome of adenovirus (AV) types 1, 2, and 5 is known to be very variable as evidenced by the great number of genome types described (36, 61, and 35 for AV1, 2, and 5 respectively). Physical maps were constructed for nearly all of the restriction or R-variants by biochemical methods and by adapting restriction fragments of defined molecular weight. The alterations found with seven DNA restriction endonucleases were mapped on the genome. Altered restriction sites, found for the genome types of each of the three serotypes, appeared to be randomly distributed over the genome, although mutations seemed to occur preferentially on distinct sites of the genome. Many strains, especially those of AV5, were genomically much more related with one another than with the prototype. This finding is compatible with the interpretation that the related strains were derived from an unknown common ancestor. PMID- 8712921 TI - HA1 domain of influenza A (H3N2) viruses in Finland in 1989-1995: evolution, egg adaptation and relationship to vaccine strains. AB - The HA1 gene sequences of 22 MDCK cell-derived influenza A (H3N2) strains, ten of their egg-derived counterparts and three vaccine strains were determined. Antigenic and sequence differences between the epidemic and vaccine strains were recorded, most striking in 1992/93; a minority of the amino acid differences in 1989-95 was involved in egg-adaptation. Changes in the assortment of amino acid substitutions produced during egg-adaptation of field strains may account for the difficulty encountered in isolating these viruses in embryonated eggs. Six revertant amino acids, characteristic of field strains prevalent in 1969-71 were recorded in 1994/95. Their genome sequence was interpreted to have been maintained over the interval years among low abundant sequences of the viral quasispecies. Potential changes of carbohydrate moieties were recorded in two glycosylation sites, suggesting that oligosaccharides at these sites are not necessarily advantageous for the H3N2 subtype virus currently. PMID- 8712922 TI - Genetic basis of the neurovirulence of type 1 polioviruses isolated from vaccine associated paralytic patients. AB - We examined four type 1 polioviruses isolated from the stools of patients with vaccine-associated paralytic poliomyelitis in China. All of these isolates were shown to be Sabin derived viruses by restriction fragment length polymorphism assay after polymerase chain reaction and by sequencing of the viral genome encoding the viral coat protein, VP1. However, the same analysis of the 3D coding region suggested that two of the four isolates had the sequence of wild type poliovirus in the tested region. Furthermore there were also point mutations in the 5' non-coding region. One was a single base change from U to C at nucleotide position 525, and the other three were from G to A at position 480. All the four strains were more neurovirulent that Sabin type 1 virus in transgenic mice with human poliovirus receptor gene. The data showed that the nucleotide positions of type 1 poliovirus which were identified to be in favor of the high neurovirulence were indeed changed during natural transmission, and suggested that the point mutation alone or a recombination of the vaccine type with wild type genome results in an acquisition of neurovirulence. PMID- 8712923 TI - Immunogenicity and relative attenuation of different vaccinia-rabies virus recombinants. AB - Immunogenicity and relative attenuation were examined for the following Tian Tan strain vaccinia-rabies recombinant viruses: 1) NGc-1, which coexpresses the glycoprotein (G) and nucleocapsid protein (N) of the rabies virus Challenge Virus Standard (CVS) strain; 2) Nc-1, which expresses the CVS N; 3) Gc-2, Gc-3, Gc-4, and Gc-5, which express CVS G via promoters from different vaccinia strains or from different vaccinia genome loci; 4) Ga-1, which expresses the G of rabies virus strain aG; and 5) Gas-1; which expresses the carboxyltruncated G ectodomain (Gs) of strain aG. All but Nc-1 and Gas-1 induced rabies virus neutralizing antibodies (VNAs) and protected groups of mice at very high frequencies from intramuscular (IM) or intracranial (IC) challenge with CVS or SW1 Shanghai dog street rabies virus (SRV); Nc-1 and Gas-1 were partly protective, more frequently against IM challenge. NGc-1 and Gc-5 appeared to induce high levels of VNAs sooner after immunization than the other constructs in mice. Relative attenuation assessed by IM infection of neonatal mice, IC infection of adult mice, and intradermal infection of rabbits with varying doses was best for NGc-1. All the recombinants were at least 100-fold more attenuated than the parent, Tian Tan vaccinia virus. Gc-2, Gc-3, Gc-4, Gc-5, and NGc-1 induced VNAs after immunization of dogs, and a subset of VNA-positive animals vaccinated with NGc-1 or Gc-3 were protected against an otherwise lethal IM injection of SRV at 21 days after vaccination. PMID- 8712924 TI - Determination of the taxonomic position and characterization of yam mosaic virus isolates based on sequence data of the 5'-terminal part of the coat protein cistron. AB - The sequences of the N-terminal part of the coat protein cistron from six isolates of yam mosaic virus (YMV-TOG, YMV-COT, YMV-12, YMV-CAR, YMV-BU1 and YMV BU2) were determined. The analysis of the deduced amino acid sequences revealed the presence of consensus motifs characteristic of the potyvirus genus supporting the classification of YMV as a potyvirus member. The alignment of the N-terminal part of the coat protein of YMV-TOG, YMV-COT, YMV-12 and YMV-CAR showed that they were identical in size (152 aa) while YMV-BU1 and YMV-BU2 were shorter (140 aa) due to a deletion of 12 aa. These amino acid sequences exhibit an overall sequence identity ranging from 70.4% to 97.4% while the identity level with the other potyviruses sequenced in the considered region is below 50%, confirming that YMV is a distinct member of the potyvirus genus. The detailed analysis of the amino acid sequence alignment and of the identity levels observed between the N-terminal part of the coat protein of the six YMV isolates lead us to suggest that they have to be considered as distantly related strains of YMV rather than closely related but distinct viruses. PMID- 8712925 TI - Prevalence of G and P serotypes among equine rotaviruses in the faeces of diarrhoeic foals. AB - Variant types of VP4 and VP7 gene segments of faecal rotaviruses from diarrhoeic foals were identified by restriction endonuclease digestion of reverse transcription/polymerase chain reaction (RT/PCR) products. The variants observed were correlated with serotypes by determination of the sequence of representative RT/PCR products (entire coding sequence for VP7 and the VP8 region of VP4) and comparison to published sequences of equine G and P serotype genes. Both G and P serotypes could be predicted for 95/116 (82%) strains, P serotype only for a further 8 (7%) strains and G serotype only for 1 (1%) strain. All characterised strains belonged to the same P serotype, P12, although minor sequence variations were observed. Of those strains able to be assigned to G serotypes, 84/96 (87.5%) belonged to serotype G3A, and 12/96 (12.5%) belonged to serotype G14. Comparison of G serotyping by ELISA to the RT/PCR method showed that serotyping equine rotaviruses by currently available ELISA methods was prone to error. This study establishes the restricted serotypic diversity of equine rotaviruses, and the significance of serotype G14. PMID- 8712926 TI - Phenotypic expression of HA-NA combinations in human-avian influenza A virus reassortants. AB - Human-avian and human-mammalian influenza A virus reassortant clones with the neuraminidase (NA) gene of the A/USSR/90/77 (H1N1) strain and hemagglutinin (HA) genes of H3, H4 and H13 subtypes had been shown in an earlier publication to produce low HA yields in the embryonated chicken eggs. The low HA titers had been shown to be due, at least in part, to the formation of virion clusters at 4 degrees C; the clustering was removed by the treatment with bacterial neuraminidase [Rudneva et al., Arch. Virol (1993) 133: 437-450]. By serial passages of the reassortants in chick embryos non-aggregating variants were selected: the variants produced HA titers of the same order as A/USSR/90/77 parent virus. The assessment of the virus yields by the analysis of the partially purified virus preparations from fixed volumes of the allantoic fluid revealed that actual virion yields of the initial reassortants were lower than the yields of their passaged variants or of the parent viruses. The passaged variant of a reassortant possessing the HA gene of A/Duck/Ukraine/1/63 (H3N2) virus differed from the original (non-passaged) reassortant and from the parent A/Duck/Ukraine/1/63 virus in the reaction with a panel of monoclonal antibodies against H3 hemagglutinin. The data suggest that some HA-NA combinations may lead to an incomplete functional match between HA and NA and to the formation of low yield reassortants, thus representing a possible limiting factor in the emergence of new HA-NA combinations in natural conditions. PMID- 8712927 TI - Usefulness and limitation of phylogenetic analysis for hepatitis C virus core region: application to isolates from Egyptian and Yemeni patients. AB - We report here the nucleotide sequences of the core region of HCV isolates from Egyptian and Yemeni patients and the method for classifying these HCV isolates by phylogenetic analysis. Sequence comparison suggested that the genotypes of these isolates were the same. Preliminary phylogenetic analysis of the HCV core region indicated that the genotypes of both isolates were 1c. However, an additional phylogenetic tree of the HCV core region constructed using a greater number of HCV isolates than that used in the preliminary analysis and on the basis of alignment of nucleotide sequences in an appropriate length indicated that the genotypes of these isolates were 4 and not 1c. For a more detailed analysis, the nucleotide sequences of the HCV E1 region as well as the core region for the same Yemeni patient were determined. A phylogenetic tree of the E1 region confirmed that the genotype of the HCV isolate from the Yemeni patient was 4. These data indicate that even when classifying HCV isolates using phylogenetic analysis, the misclassification would occur if care is not taken regarding the number and sequence lengths of the isolates included in the analysis. PMID- 8712928 TI - Conservation of coat protein sequence among isolates of potato mop-top virus from Scotland and Peru. AB - Coat protein gene sequences of eight isolates of potato mop-top virus from the Peruvian Andes and of three isolates from Scotland were compared. Despite wide geographical separation, there was little sequence variation among all isolates. PMID- 8712929 TI - Structural and functional analysis of the S proteins of two human coronavirus OC43 strains adapted to growth in different cells. AB - The receptor-binding activity of strain CU (grown in MDCK I cells) and of strain VA (adapted to Vero cells) of human coronavirus OC43 was analyzed and compared with the binding activity of bovine coronavirus (BCV) and of the OC43 strain provided by the American Type Culture Collection (AT). Results obtained with resialylated erythrocytes indicated that the ability of the viruses to recognize 9-O-acetylated sialic acid in an alpha 2,6-linkage decreased in the following order: AT > CU > BCV > VA. Only minor differences were observed with respect to the alpha 2,3-linkage. The amino acid sequence of the S protein of strain CU and VA was derived from the nucleotide sequence of the cloned gene. Strain VA differed from strain CU in 34 positions, 18 in the S1 and 16 in the S2 subunit. PMID- 8712930 TI - Determination of the 5' and 3' terminal noncoding sequences of the bi-segmented genome of the avibirnavirus infectious bursal disease virus. AB - Terminal sequences of the bi-segmented dsRNA genome of 3 different strains of infectious bursal disease virus (IBDV) were analyzed by the rapid amplification of cDNA 5' ends (5'RACE) procedure. Both segments are 85% homologous in a 32 nucleotide sequence comprising the 5' end, whereas the 3' end has a conserved pentamer. Comparison to published terminal sequences of other IBDV strains revealed high conservation between the two segments but more serotype-specific nucleotide changes (5 on segment A and 3 on segment B) in the 5' noncoding region compared to the 3' noncoding region (none on segment A and 1 on segment B). PMID- 8712931 TI - Site-specific mutations in the NS2 protein of epizootic haemorrhagic disease virus markedly affect the formation of cytoplasmic inclusion bodies. AB - The importance of a conserved amino acid motif in the nonstructural protein NS2 of different orbiviruses was investigated with regard to virus inclusion body (VIB) formation. A number of epizootic haemorrhagic disease virus NS2 deletion and substitution mutants were prepared and expressed as baculovirus recombinants. Deletion of the motif or substitution of at least three residues within the region, had a detrimental effect on VIB formation in insect cells. Furthermore, these NS2 mutants were not complexed with single-stranded RNA in infected cells and appeared to be cytotoxic. Cumulatively, the results suggest that this motif is an essential structural determinant for NS2 function. PMID- 8712932 TI - Deletion of the carboxy-terminus of herpes simplex virus type 1 (HSV-1) glycoprotein B does not affect oligomerization, heparin-binding activity, or its ability to protect against HSV challenge. AB - A recombinant vaccinia virus designated VgBt which expresses a truncated secreted herpes simplex virus gB (gBt) was constructed and compared to V11gB, a vaccinia recombinant previously studied which expresses gB exclusively on the surface of infected cells. Indirect immunofluorescence assay (IFA) revealed that gBt was strongly associated with the surface of infected cells despite being released slowly into the cell culture medium. Both gB and gBt existed as oligomers, and both membrane bound and secreted forms of gBt exhibited heparin-binding activity. In protection studies VgBt and V11gB conferred equivalent protection against both homologous (HSV-1) and heterologous (HSV-2) challenge with HSV. PMID- 8712933 TI - Accumulation kinetics of CMV RNA 3-encoded proteins and subcellular localization of the 3a protein in infected and transgenic tobacco plants. AB - The complete nucleotide sequence of RNA 3 of a Spanish isolate of cucumber mosaic virus (CMV-24) has been determined. The encoded putative cell-to-cell movement protein (3a protein) and the coat protein are 279 and 218 amino acids long, respectively. The 3a protein was expressed in Escherichia coli using the vector pT7-7 and was used to raise an immunoserum. We have followed the time course of accumulation of the 3a protein, in parallel to that of the coat protein, and its subcellular localization as a function of time after CMV-24 infection on tobacco plants. The maximum accumulation level of the 3a protein was reached at early stages of infection, being detected in the cytosolic and the cell wall fractions. At later stages of infection, a decline in accumulation levels of the 3a protein was observed, and the protein was essentially associated with the cell wall fractions. These data were corroborated by immunocytochemistry performed in both infected and 3a-expressing transgenic tobacco plants. PMID- 8712934 TI - [Immunohistochemical characterization and ultrastructural features of cervical squamous cell carcinoma of ectocervical and endocervical (cambial cell or metaplastic) genesis]. AB - Among 199 invasive squamous cell carcinomas immunophenotype of ectocervical carcinoma (true squamous cell one) was found in 18 tumors. Immunophenotype of metaplastic, endocervical squamous cell carcinoma was established in 181 cases. There was a pronounced expression of cytokeratins N 1, 2, 10, 14 and negative or weak expression of N 8 and 17. Ultrastructural features of this carcinoma type were desmosomes on microprocesses, abundant bundles of cytoplasmic tonofibrils. A high level of cytokeratins N 8 and 17, carcinoembryonic antigen was found in squamous cell endocervical carcinoma; at the same time keratins N 1, 2, 10 and 14 were weakly expressed, there was also a characteristic ultrastructure of intercellular contacts (desmosomes on flat surfaces) and microvilli. The results allow to use immunohistochemical and ultrastructural methods for differential diagnosis of squamous cell carcinoma originating from different sources. PMID- 8712935 TI - [Current methods for studying the functional morphology of apud cells]. AB - The paper reviews methods for studying functional morphology of endocrine cells histochemistry and immunohistochemistry, radioautography and methods of ultrastructural verification of secretory granules. Methodological approach is illustrated by the authors' results on the influence of ionizing radiation and tumor growth on the cells of the diffuse endocrine system. PMID- 8712936 TI - [Differential immunohistochemical diagnosis of tumors of the human central nervous system using monoclonal antibodies]. AB - 67 brain tumors were studied using monoclonal antibodies against glial fibrillary acid protein (GFAP), neurofilaments, proteoglycan, laminin and vimentin. All the astrocytic tumors gave positive reaction to GFAP, but anaplastic tumors of this type were less reactive. Medulloblastomas when treated with GFAP showed two cell lines, astrocytic and neuronal. A positive reaction to laminin and proteoglycan was observed in the vessels of neuroepithelial tumors. Anaplastic astrocytomas were more reactive to vimentin. PMID- 8712937 TI - [Apoptosis and its role in the mechanisms of growth regulation of tumor cells with multiple drug resistance]. AB - Development of apoptosis was followed up in the cell line LIM-1863 of human colon carcinoma and in the same cell line with multiple drug resistance (MDR) related to the expression of gene mdr I and hyperproduction of protein P-glycoprotein 170 encoded by this gene. The number of cells with histological and ultrastructural features of apoptosis increased with acquirement by tumor cells of typical MDR. The enhancement of apoptosis in tumor cells with MDR results probably from the increase of cells with signs of terminal differentiation which is one of apoptosis inducers. Mitotic activity in both lines did not change essentially, but the original line had a more rapid growth than its analogue with MDR. Elimination of cells through apoptosis may be the cause of slower growth of the cell line with apoptosis. The rate of tumor growth depends on the balance between proliferative activity and apoptosis. PMID- 8712938 TI - [Immunohistochemical study of the expression of proliferating cell nuclear antigen in astrocytic gliomas of the cerebral hemispheres]. AB - Proliferating cell nuclear antigen (PCNA) was studied in 136 biopsies of brain astrocytic gliomas (AG) subdivided according to the degree of their malignancy into 3 groups: astrocytomas, anaplastic astrocytomas and glioblastomas. There were statistically significant differences in labelling index (LI) between these groups: 5.2, 34.1 and 49.5%, respectively. Mean LI of PCNA in the anaplastic astrocytomas was 6.5 times higher than in benign astrocytomas. Expression of PCNA in the tumor endotheliocytes was observed in 11 of 24 histologically benign astrocytomas and LI in these tumors was considerably higher than in the astrocytomas without endothelial expression. This indicates that proliferative processes in the tumor cells of AG and in the endotheliocytes of tumor vessels are interconnected. PMID- 8712939 TI - [Ultrastructural pathology of the lymph nodes in tick-borne encephalitis]. AB - Lymph nodes of humans and experimental animals with tick-borne encephalitis (TBE) were studied using electron microscopy. TBE virus reproduction occurred in macrophages, reticular and plasma cells. Severe organelle destruction was found in the above cells and lymphocytes. The results suggest that macrophage, reticular and plasmatic cell death is due to the virus reproduction, while lymphocyte death is due to the toxic effect. It is evident that the outcome is determined by pathological process in lymphoid organs. PMID- 8712940 TI - [Endotoxin-induced injury to the endothelium]. AB - Literature and own data on endotoxin- induced injuries to endothelium are reviewed. It is shown that endotoxin can cause, hyperactivation of granulocytes, activation of complements, local endothelial injuries and some increase of vascular cell wall permeability, oxidation of low-density lipoproteins (LDL) and LDL-LPS complexes, binding of LPS with high-density lipoproteins (HDL) and some decrease of HDL ability to bind cholesterol, stimulation of endothelial and smooth muscle cell replication in local injuries to vessel wall. Low doses of endotoxin were found in blood plasma and on granulocytes surface in healthy and sick subjects. It is concluded that intestinal microflora endotoxin may play an essential role in pathogenesis of atherosclerosis. PMID- 8712941 TI - [Cardiomyocyte cytoskeleton in dilated cardiomyopathy and ischemic heart disease]. AB - Summary. Vinculin, the protein of cardiomyocyte cytoskeleton in the myocardium of patients with a dilated cardiomyopathy and ischemic heart disease, was revealed by means of monoclonal antibodies and by a method of indirect immunofluorescence. Cytoskeleton of patients with dilated cardiomyopathy is disorganized, hypertrophic and contains greater amounts of vinculin than cytoskeleton of normal cardiomyocytes. This most likely negatively influences cell contractile capacity. The vinculin content was also higher in cardiomyocyte cytoskeleton of patients with ischemic heart disease but at a less degree than in patients with dilated cardiomyopathy. PMID- 8712942 TI - [Ultrastructure of intermitochondrial contacts of human cardiomyocytes in alcoholic cardiomyopathy and ischemic heart disease]. AB - Intermitochondrial contacts in human cardiomyocytes are revealed. In alcoholic cardiomyopathy they have the same morphological features as those previously described in the cardiomyocytes of vertebrates. The ultrastructure of these contacts in patients with circulation deficiency in ischemic heart disease is altered. This alteration of the ultrastructure may represent a direct reaction of the cardiomyocyte mitochondrial system to the change in the level of energetic metabolism. PMID- 8712943 TI - [Low-energy laser therapy of chronic gastritis in children]. AB - Morphological and morphometric study of gastric mucosa was performed before and after laser therapy in children with chronic gastritis. Features of childhood gastritis are provided and laser therapeutic action in different after treatment periods is characterized. PMID- 8712944 TI - [Mathematical modeling of the dynamics of structural thyroid parameters in thyroid disease with immune disorders]. AB - The material of the study was surgical thyroid biopsies from 126 patients, males and females, aged 14-69 years. 5 groups were studied depending upon the histological picture and the degree of lymphoid infiltration. Thyroids from 10 persons without thyroid pathology who died a violant life served as control. As a result of the factor analysis 5 main factors were distinguished. The first one is lymphoid infiltration, the 2nd and 3rd ones-factors of thyroid epithelium characterizing the structure and the size of thyrocytes, the 4th and the 5th factors characterize the structure of the thyroid as an organ. The analysis confirmed lymphoid infiltration and thyrocyte dimensional parameters are the most important morphological criteria in the diagnosis of thyroid pathology with immune disturbances. Six equations are derived allowing to classify new cases of thyroid pathology. PMID- 8712945 TI - [Pathology of congenital bromine deficit (experimental observation)]. AB - Pathology of congenital bromine deficiency is described in a she-goat whose mother was kept for two years on a bromine deficient diet. Anomalies of the bones and joints of the anterior limbs, disturbances of carbohydrate, lipid and mineral metabolism in the form of liver, kidney and lymph nodes glycogenosis, liver fat degeneration, haemosiderosis of the liver, spleen and lungs were found. Moderate thymomegaly with hyperplasia of the cortex and an increase in the number of Hassal bodies, microcystic ovary transformation, adrenal adenomatosis, cheilitis, flossitis, focal proliferative extracapillary glomerulonephritis were also observed. PMID- 8712946 TI - [Issue of certification of clinical pathologists (specialists, doctor of second, first and highest category)]. PMID- 8712947 TI - [Analysis of mortality in outpatient departments]. AB - Outpatient mortality has been analyzed basing on the study of 470 case histories and autopsy protocols for the period 1984-1993. Cardiovascular diseases predominated among the death causes (86.1%). Ischemic heart disease accounted for 87.4% deaths due to the above diseases, Clinical and post-mortem diagnosis disagreed by the prior disease in 8.9%, by location of the tumor in 0.6% of cases. PMID- 8712948 TI - [Puncture cytologic diagnosis of thyroid diseases: potentialities and limitations of the method]. AB - The role of fine-needle aspiration biopsy in cytological diagnosis of thyroid diseases is reviewed. This method allows not only to determine whether the condition is benign or malignant, but also diagnose nosological variants of thyroid pathology. The difficulties lie in the diagnosis of combined thyroid pathology, differential diagnosis of follicular adenoma with follicular carcinoma, lymphoma with autoimmune thyroiditis, follicular tumor with adenomatous goiter. PMID- 8712949 TI - [Role of gram-negative bacterial endotoxins in infectious and non-infectious diseases]. AB - The role of gram-negative bacteria endotoxin in infectious and non-infectious pathology is reviewed. It is shown that endotoxin may induce some injuries to the lungs, liver, kidney and blood vessels. It is suggested that intestinal microflora endotoxin may take part in the development of many pathological processes. The basis of the endotoxin effect is its capacity to interact with cell membrane. Intravascular blood coagulation and endotoxin shock resulting in the patient's death may occur at high doses of endotoxin. PMID- 8712950 TI - Small milestones in my artificial heart research. PMID- 8712951 TI - Cytoplasmic calcium levels and membrane fluidity of platelets in contact with polyether-polyamide multiblock-copolymer surfaces. AB - Cytoplasmic calcium levels and the membrane fluidity of rabbit platelets stored in mini blood bags of crystalline-amorphous microstructured polymers (polyether polyamide multiblock-copolymers) were studied. Fluorescent dye (Fura 2 or 1,6 diphenyl-1,3,5-hexatriene)-loaded platelet suspensions were stored at 37 degrees C for 1 h in the blood bags, and metabolic changes in the platelets during storage were evaluated by the fluorescent spectroscopic technique. The surfaces of poly(vinyl chloride) and polyolefin elastomers, which are used for commercially available blood bags, enhanced the progress of platelet metabolism; i.e., there was a dramatic decrease in membrane fluidity and an increase in [Ca2+]i. Furthermore, the decrease in membrane fluidity was observed prior to the increase in [Ca2+]i. These results suggest that the decrease in membrane fluidity of platelets in contact with polymer surfaces can be the dominant stage in the activation of these platelets. In contrast, the surfaces of polyether-polyamide multiblock-copolymers exhibited few changes in either membrane fluidity or [Ca2+]i levels. These results suggest that the platelets in contact with the crystalline-amorphous microstructured copolymer surfaces can be inert and inactivated in terms of the prevention of a decrease in membrane fluidity. PMID- 8712952 TI - Marine propellers: the latest topics. AB - The impeller of the axial flow blood pump in an artificial heart is essentially based on the same principle as a marine propeller. Impellers designed for artificial hearts and marine propellers have a number of points in common. Decreased cavitation and relieved fluctuation load are only representative of them. As for a distinct concept of pressure distribution, the inverse method could be very useful. Skew may led to a more mild and natural character in the blood. Highly skewed blades and super elastic blades have the potential to decrease the burden on the entire circulatory system. This paper will address the main points and latest issues in propeller design concluding with a discussion of the implications of these issues for blood pump impellers. PMID- 8712953 TI - Skeletal muscle ventricles: frontiers in 1995. AB - Skeletal muscle ventricles (SMVs) constructed from electrically conditioned latissimus dorsi muscle (LDM) may become an alternative for assisting the failing heart. Left and right heart circulatory assist using SMVs has been performed successfully in both acute and chronic animal models. The configurations used to connect SMVs to the circulation have included a left atrium to aorta bypass, a left ventricle apex to aorta bypass, aortic counterpulsators, a cavopulmonary bypass, and a right ventricle to pulmonary artery bypass. One SMV used as an aortic counterpulsator functioned effectively in the circulation for more than 27 months. Recent application of the pericardium to the SMV as an inner layer and design changes in the connection of the SMV to the circulation have reduced the risk of thrombus formation and SMV rupture. Although several problems have yet to be solved, the goal of the SMV as a permanent circulatory assist device without the limitation of an external power source seems within reach. PMID- 8712954 TI - Current status of extracorporeal membrane oxygenation for severe respiratory failure. AB - Extracorporeal membrane oxygenation (ECMO) for respiratory failure was reviewed. ECMO progressed from laboratory research to initial clinical trials in 1972. Following a decade of clinical research, ECMO is now standard treatment for neonatal respiratory failure refractory to conventional pulmonary support techniques worldwide. The application of neonatal ECMO has been extended with improved outcome to premature and low birth weight infants as well as older children and adults. As of July 1994, 9,258 neonates, 754 pediatric, and 130 adult patients with respiratory failure treated with ECMO were entered in the registry of the Extracorporeal Life Support Organization (ELSO). Overall survival rates were 81% in neonates, 49% in pediatric, and 38% in adult patients. Recently the adult and pediatric populations treated with ECMO have increased rapidly, and the outcome has improved significantly. PMID- 8712955 TI - Long-term animal survival with an implantable axial flow pump as a left ventricular assist device. AB - We are developing an axial flow blood pump with Nimbus Inc. (AxiPump). For in vivo evaluation the AxiPump has been used as a left ventricular assist device with a left ventricular and descending aorta cannulation and implantation in a small pocket on the left lateral abdominal wall just posterior to the costal margin. Electrical and flow probe leads exit the body transcutaneously. A purge line that delivers the purge fluid for lubrication of the seal between the rotor and stator bodies in the purge fluid bearing system is tunneled with the other leads. Following acute animal studies, 3 animals have been supported for over 1 month with this AxiPump system. All laboratory results were within normal limits except during a recovery period from surgical damage. Hemolysis was not a serious problem. In the first case, the purge system failed at 28 days, and in the second and third cases, the nonpurge bearing system worked well for 57 and 52 days, respectively. Bearings are still under development in this kind of pump. However, this success encourages us to improve the AxiPump as a long-term assist device. PMID- 8712956 TI - A new magnetically suspended centrifugal pump: in vitro and preliminary in vivo assessment. AB - To overcome problems derived from the shaft within the conventional centrifugal pump, we have been developing a new centrifugal pump, namely a magnetically suspended centrifugal pump (MSCP), which has no shaft and operates as a noncontacting and bearingless pump. The impeller is suspended magnetically between the magnetic bearing and the driving motor. Hemolysis tests were performed in comparison with the Biopump (BP80, BioMedicus). The index of hemolysis (IH) was significantly lower in the MSCP than in the Biopump. In addition, a smaller gap in the MSCP induced lower hemolysis. In preliminary studies using mongrel dogs, the layer of thrombus adherent to the impeller was observed in a few hours, which impaired the pumping efficiency. However, by using an impeller coated with silicone, no aggregations of platelets or fibrin on the impeller were observed in 24 h of continuous pumping. In conclusion, the MSCP had a gentler influence on blood cells than the Biopump, and the impeller coated with silicone may contribute to the long-term pumping of the MSCP. PMID- 8712957 TI - A quantitative visualization study of flow in a scaled-up model of a centrifugal blood pump. AB - A quantitative flow visualization study of a scaled-up model of a centrifugal blood pump was performed. Since the size of the scaled-up model was three times as large as the original pump under development, and the kinematic viscosity of the saline solution used as the working fluid was approximately one-third that of the blood, we obtained a similar flow at one twenty-seventh the angular velocity of the original pump. The flow was visualized by seeding the saline solution with neutrally buoyant particles and by illuminating the model with a laser light sheet. Since the gap flow behind the impeller is important for thrombus formation, it was recorded by a high-speed video camera, and the velocity field was evaluated automatically by particle tracking velocimetry. It was shown that in the gap behind the impeller there existed a region where the velocity profile was almost flat which can be called a core region. The results indicated the effectiveness of the present visualization technique for centrifugal blood pumps. PMID- 8712958 TI - Comparison studies of major organ microcirculations under pulsatile- and nonpulsatile-assisted circulations. AB - In these comparison studies, we examined changes in major organ microcirculations during circulatory support using pulsatile and nonpulsatile pumps. Acute myocardial infarction was created by left anterior descending (LAD) branch ligation. After the animals in these studies fell into cardiogenic shock, they were supported by mechanical assist devices: a pulsatile ventricular assist device (VAD) in 4 pigs, a nonpulsatile VAD in 4 pigs, and an intraaortic balloon pumping (IABP) plus nonpulsatile VAD in 4 pigs. Each group was supported for 3 h with an identical mean aortic pressure being maintained. As for parameters, the stomach mucosa, liver regional blood flow, stomach mucosal pH, and arterial blood keton body ratio (AKBR) were measured. Both regional blood flow and AKBR increased in the pulsatile group as compared with the nonpulsatile group. There were no differences in the stomach mucosal pH among the 3 groups. These results suggest that pulsatile assist rather than the nonpulsatile assist plays a significant role in the recovery of deteriorated splanchnic organs due to cardiogenic shock. PMID- 8712959 TI - Influence of ventricular fibrillation on sympathetic nerve activity under biventricular bypass circulation. AB - Autonomic nerves play an important role in circulatory control. The characteristics of sympathetic nerve activity (SNA) during artificial circulation, however, are not well understood. In this study, we examined the influence of natural heart beating on the renal SNA (RSNA) using pulsatile 100% biventricular assistance and electrically induced ventricular fibrillation in anesthetized goats, whose heart rate and size are similar to the human heart's. Following induction of the ventricular fibrillation, the pulse-synchronous discharges in the RSNA did not change their periodicity and quantity significantly. It was concluded that the beating of the heart had little influence on the RSNA and was presumably not essential for maintaining nervous control of circulation as long as the circulation was maintained by an artificial heart. PMID- 8712960 TI - A percutaneously accessible pulsatile left ventricular assist device: modified assist device type 5. AB - To provide percutaneous access, a new circulatory assist system was developed. We call this newly developed system the modified assist device (MAD). The system is composed of a sac-type blood pump and cannula. Inflow and outflow valves are mounted in the apex and at the side wall 10 cm from the apex of the cannula, respectively. During systole, the blood is sucked from the left ventricle through the inflow valve of the cannula connected to the blood pump, and during diastole, the blood is ejected to the root of the aorta through the outflow valve. In vitro and in vivo evaluations of the pump performance were performed. The maximum flow rate of 1.9 L/min was obtained in the mock circulatory system. In our animal experiment, effective systolic unloading and diastolic augmentation were observed by activation of this system during regular sinus rhythm. In conclusion, the MAD 5 is thought to be percutaneously accessible and increases systemic and coronary flow. PMID- 8712961 TI - In vitro and in vivo testing of an implantable motor-driven left ventricular assist device. AB - A totally implantable motor-driven left ventricular assist device (LVAD) has been developed and tested. The performance of this LVAD was tested in a mock circulatory system. This pump provided 8 L/min of output against a mean afterload of 120 mm Hg with a filling pressure of 20 mm Hg when the pump was operated in the fill/empty mode. The right and left pumps were tested in a mock loop. The right pump afterload was kept in the range from 23-32 mm Hg. With increase in the left pump afterload, the pump power output varied from 1.64 to 2.37 W. The instantaneous motor power input varied from 22.6 to 30.6 W with the total system efficiency ranging from 6.7 to 9.4%. To date, 4 in vivo studies have been conducted for up to 12 h. Two animals survived 12 and 10 h, respectively. Termination was due to bleeding in 1 animal, vent tube obstruction in 1, and respiratory failure in 2. All animals died of technical failure. Another experiment is to be undertaken, and a newly designed cannula is now being manufactured. PMID- 8712962 TI - Linear skeletal muscle ventricle: a pilot study. AB - We have developed a new skeletal muscle ventricle (LSMV) powered by linear contraction of the latissimus dorsi muscle (LD). This consists of two bellows of different diameters joined by a connector containing a valve. The smaller bellows is connected to the left atrium with a valve, and the larger bellows is connected to the aorta. The caudal tendon of the LD is attached to the connector. LD contraction pulls the connector to compress the larger bellows and to elongate the smaller bellows. This motion closes the valve in the connector, ejects blood in the larger bellows, and draws blood into the smaller bellows. LD relaxation allows aortic blood to regurge into the larger bellows. Therefore, the LD is stretched. In acute canine experiments, the stroke work and power output of the LSMV measured 0.214 J and 0.135 W with a filling pressure of 4.3 mm Hg. PMID- 8712963 TI - Double balloon catheter for concomitant augmentation of abdominal organ perfusion during intraaortic balloon counterpulsation. AB - A newly devised double balloon catheter to augment abdominal organ perfusion concomitant with original cardiac assist during intraaortic balloon counterpulsation provided satisfactory hemodynamics and urinary output around cardiac surgical procedures in 2 patients with coronary artery disease. Some optional advantages in clinical application and problems arising in the catheter design which need to be investigated are discussed. PMID- 8712964 TI - Evaluation of intramyocardial coronary blood flow waveform during intraaortic balloon pumping in the absence or presence of coronary stenosis. AB - Our aim was to evaluate the effects of intraaortic balloon pumping (IABP) on the blood velocity waveform in the absence or presence of coronary artery stenosis. Using anesthetized open-chest dogs, the septal arterial blood flow velocities were measured with a 20 MHz 80-channel ultrasound pulsed Doppler velocimeter in the absence (n = 5) or presence (n = 3) of left main coronary artery stenosis. The blood velocity waveform was analyzed by calculating the systolic retrograde velocity integral (SR) and the diastolic antegrade velocity integral (DA). A slosh ratio was defined as SR/DA. The left anterior descending arterial flow (CBF), aortic pressure (AoP), and poststenotic distal coronary pressure (DiP) were also measured simultaneously. We compared the effect of IABP on the velocity waveforms in the absence and in the presence of coronary artery stenosis. In the absence of stenosis, IABP increased DiP during diastole and augmented DA while it also increased SR. IABP augmented the net CBF because of the greater increase in DA than SR. In the presence of stenosis, however, IABP did not increase DiP and resulted in no significant effect on the net CBF. PMID- 8712965 TI - Optimal oxygen tension conditions for functioning cultured hepatocytes in vitro. AB - With a view toward furthering the development of artificial liver systems, we have been culturing hepatocytes in vitro. The object of this research was to investigate the ideal conditions of oxygen tension for the efficient functioning of hepatocytes. Viable hepatocytes isolated from rat livers were cultured under five different oxygen tensions: 5, 10, 20, 50 and 90% O2. DNA contents, gluconeogenesis, urea synthesis, adenosine triphosphate (ATP) levels, and lipid peroxidation of hepatocytes were evaluated. Under the 5% oxygen conditions, the function of hepatocytes was very inferior and was accompanied by a low ATP level. However, hepatocytes cultured under 90% oxygen tension functioned less effectively than the control (20% O2) with elevated lipid peroxidation. The data in this study suggest that the optimum oxygen condition for cultured hepatocytes is 10 approximately 50%, and that especially under conditions of 20% oxygen tension, i.e., that of the ordinary atmosphere, hepatocytes can function most effectively. PMID- 8712966 TI - Development of a bioartificial liver with sandwiched-cultured hepatocytes between two collagen gel layers. AB - We have been developing a multiplate type of bioartificial liver (BAL) using primary cultured hepatocyte monolayers since 1987. This BAL has been shown to prolong the survival time of anhepatic dogs and rabbits. Initially, hepatocytes were cultivated on collagen-coated plates. To increase the multiplated BAL function, a sandwich cultivation method was employed in which rat hepatocytes were cultured between two collagen gel layers and were then evaluated in both stationary and perfusion cultures. In the stationary culture, the sandwich method showed a higher activity in urea synthesis than in the other culture methods (culture on a collagen coating, culture on a collagen gel, and culture between a collagen coating and a collagen gel) for 14 days. In the perfusion culture, a BAL housing cultured hepatocytes (6.5 x 10(7) cells) in the sandwich system showed urea synthesis activity ranging from 17.5 to 22.6 micrograms/2 x 10(6) cells/90 min. This activity was maintained for 5 days in the perfusion culture. The sandwich type of cultivation is applicable to the multiplated BAL. PMID- 8712968 TI - Should kidney failure patients be treated for kidney function or urenic syndrome? PMID- 8712967 TI - Function of culturing monolayer hepatocytes by collagen gel coating and coculture with nonparenchymal cells. AB - Since 1987, we have been developing a bioartificial liver (BAL) using multiplated cultured hepatocyte monolayers. With the goal of promoting hepatic functions of cultured hepatocyte monolayers, we combined the use of a collagen gel layer over the monolayers of hepatocytes and/or cocultured hepatocytes with nonparenchymal cells (NPCs). The study was divided into four groups according to culture configurations: Group 1: hepatocyte monolayer culture (control); Group 2; coculture of hepatocytes and NPCs; Group 3: hepatocyte monolayer with a overlaid collagen gel layer; and Group 4: coculture with a overlaid collagen gel layer. The culture continued for 14 days. Morphological changes and hepatic functions were evaluated by urea and albumin syntheses. The morphological status of the hepatocytes remained for 2 weeks in Groups 3 and 4. Deterioration and detachment of hepatocytes and/or NPCs started in Group 1 and 2 on the third day in culture. Significantly high urea synthesis was noted in Group 4 (p < 0.001 compared with Group 1 and 2: p = 0.0014 compared with Group 3). Although there was no significant difference in albumin synthesis among the four groups, those hepatocytes covered by the collagen gel (Groups 3 and 4) tended to secrete albumin throughout the observation period. These results indicted that the environment, although artificial (but close to the in vivo state), supplied with collagen gel and the coculture, enhanced the activities of the cultured hepatocyte monolayers. We suggest that use of cocultured hepatocytes under a collagen gel is a promising candidate for a bioreactor of multiplated BAL. PMID- 8712969 TI - Ultrasonographic assessment of the swelling of the human masseter muscle after static and dynamic activity. AB - Work-related fatigue, pain and disorders in skeletal muscles have been related to prolonged static and dynamic activity. Such contractions have been shown to impair blood flow and increase muscle thickness and fluid. In the present study the effect of static and dynamic activity was evaluated from changes in masseter thickness as a measure of oedema, simultaneously with assessment of perceived pain/discomfort and cardiovascular responses. As static activity, fourteen young healthy women bit at 15% maximal voluntary contraction on bite-force transducers in the molar regions until exhaustion or 20 min at maximum (median endurance time 7.1 min). For dynamic activity, the same individuals chewed gum unilaterally until exhaustion or 40 min at maximum (all endured 40 min) with a cycle time of 725 ms, an average load of 9.3% of maximal electromyographic activity (maxEMG) and a peak mean voltage of 54.3% of maxEMG. Muscle thickness was measured by ultrasonography at the mid-portion of the ipsilateral masseter. Immediately after exercise, muscle thickness was significantly increased, more after static (14.0%) than dynamic (8.6%), and returned to pre-exercise values after 20-min recovery. Visual analogue scales (VAS) revealed the concomitant occurrence of pain (static 11.9 VAS%; dynamic 5.9 VAS%), and discomfort (static 8.1 VAS%; dynamic 5.9 VAS%), and both sensations decreased to pre-exercise values after 20-min recovery. Systolic blood pressure increased significantly, more during static (12.5%) than dynamic activity (4.3%), whereas heart rate rose significantly only during dynamic exercise (13.3%). Hence, activity was associated with muscular swelling and pain, and, despite the relatively small size of the masticatory muscles, also with general cardiovascular responses. PMID- 8712970 TI - Human salivary glucose analysis by high-performance ion-exchange chromatography and pulsed amperometric detection. AB - The aim of this project was to develop an analytical procedure with the required level of sensitivity for the determination of glucose concentrations in small volumes of unstimulated fasting whole saliva. The technique involves high performance ion-exchange chromatography at high pH and pulsed amperometric detection. It has a high level of reproducibility, a sensitivity as low as 0.1 mumol/l and requires only 50 microliters samples (sensitivity = 0.002 pmol). Inhibition of glucose metabolism, by procedures such as collection into 0.1% (w/v) sodium fluoride, was shown to be essential if accurate results are to be obtained. Collection on to ice followed by storage at -20 degrees C was shown to be unsuitable and resulted in glucose loss by degradation. There were inter- and intraindividual variations in the glucose concentration in unstimulated mixed saliva (range; 0.02-0.4 mmol/l). The procedure can be used for the analysis of other salivary carbohydrates and for monitoring the clearance of dietary carbohydrates from the mouth. PMID- 8712971 TI - Changes in the fibre arrangement of the rat incisor periodontal ligament in relation to various loading levels in vitro. AB - The relationship between the fibre arrangement of the periodontal ligament and the load-deformation behaviour was investigated at various loading levels. Transverse sections of the rat incisor were loaded in the eruptive direction in vitro and the deformation fixed at predetermined loads. Sections were prepared at these deformation levels. The periodontal ligaments were examined by polarized light and scanning-electron microscopy. At the initial ("toe') part of the load deformation curve, the periodontal fibres were gradually pulled and bent towards the direction of loading; their wavy pattern and periodic dark and bright bands became indistinct. At the next, linear part of the curve, the running direction of the fibres changed gradually until they were straightened and stretched. At the yielding part of the curve, the periodontal fibres began to rupture. Ruptured fibres adhering to the bone surface returned to their original obliquity and showed periodic dark and bright bands. The fibrous components of the rat incisor periodontal ligament thus transmit forces to bone at the linear part of the curve when the tooth is axially loaded. PMID- 8712972 TI - Stimulation of murine tooth development in organotypic culture by the neurotransmitter serotonin. AB - Serotonin (5-hydroxytryptamine; 5-HT) uptake sites are transiently expressed in craniofacial epithelia and mesenchyme, including the tooth germ, during mouse embryogenesis. Based on malformations and patterns of cell proliferation and death in cultured mouse embryos exposed to 5-HT uptake inhibitors, it has been hypothesized that 5-HT acts as a dose-dependent morphogenetic signal for craniofacial development. The present study was designed to investigate the effect of 5-HT on tooth-germ formation in serum-free mandibular explant cultures prepared from embryonic day-13 (plug day = embryonic day-1) mouse embryos. In the absence of serum or a 5-HT supplement, tooth germs develop only to the bud stage in these cultures. When explants were cultured for 8 days in a defined medium supplemented with 5-HT, late bell-stage tooth germs were stimulated to develop in a dose-dependent manner. This effect was reversed by addition of the 5-HT uptake inhibitor fluoxetine. Anti-5-HT immunocytochemistry demonstrated specific uptake of 5-HT by developing tooth germ and mandibular epithelium, which could also be blocked by fluoxetine. These results suggest that 5-HT may regulate dental differentiation, and that intracellular uptake is required for this action. PMID- 8712973 TI - Oral colonization by more than one clonal type of mutans streptococcus in children with nursing-bottle dental caries. AB - By ribotyping the genetic diversity of mutans streptococci in six 1.5-3-yr-old children with nursing-bottle caries and in six caries-free, age-matched children and in their mothers was examined. The proportion of mutans streptococci in the dental plaque of the children and their levels in the saliva of the mothers were also examined. For ribotyping, chromosomal DNA of isolates obtained from the plaque of the children (3-12 isolates per child) and from the saliva of the mothers (4-13 isolates per mother) was digested with restriction endonuclease HindIII. The DNA fragments were hybridized to the plasmid pKK3535 which contains the rRNA operon of the Escherichia coli chromosome. The results showed that children with nursing-bottle caries exposed to frequent consumption of sucrose had a high proportion of mutans streptococci in plaque and four of them were colonized with more than one ribotype, whereas caries-free children had a low proportion of mutans streptococci in plaque and only one of them harboured more than one ribotype. Mothers of children with nursing bottle caries had similar levels and numbers of ribotypes of mutans streptococci in saliva as the mothers of the caries-free children. In both child groups, mothers were probably the main source of infection with mutans streptococci. Thus, children with nursing-bottle caries were not only heavily infected with mutans streptococci but also often colonized with more than one clonal type. In the child's acquisition of such clones, frequent sugar consumption may have an important role. PMID- 8712974 TI - Natural bites, food consistency and feeding behaviour in man. AB - Complete feeding sequences (bite to terminal swallow) were recorded in 11 individuals (Class I occlusions) feeding on apple, banana and biscuit (hard cookie) to establish whether jaw movements characteristic of the stage I transport, chewing, stage II transport and swallowing processes found in non human mammals could be identified in man. All participants took large natural bites (known weight) of each food, but seven were supplied standard 6-g pieces of apple. Jaw movements (three axes) were recorded using a locally calibrated Sirognathograph. Discrete stage I transport movements (bite moved from incisors to postcanines), were found in most records, but least frequently for banana. The number of chewing cycles before the first in-sequence swallow increased in the order: banana, ( = ) apple without peel, apple with peel, biscuit. Chewing cycles showed subtle changes in gape profile but significant variation in mediolateral movement cycle to cycle. Morphologically distinct stage II transport cycles could not be identified. Swallows occurred intermittently during chewing (79% of all sequences, n = 146) with cycle durations in excess of 1 s in contrast to the range for chewing (0.6-0.7 s). Almost all sequences ended with a period, clearance, of highly irregular jaw movements often including swallows. Clearance has not been described in other mammals. Food consistency affected feeding behaviour. The "chew/swallow' ratio (total number of chews/total number of swallows) increased with food hardness. The number of chewing cycles before the first in-sequence swallow also increased. Food type significantly affected sequence duration. The duration of clearance was also affected by bite weight and food hardness. It is concluded that initial food consistency determines the number of chewing cycles before the first swallow and overall sequence duration. The change in the pattern of jaw movement in both the vertical and mediolateral directions cycle to cycle suggests continuous sensory modulation of the motor output to the mandibular musculature. PMID- 8712975 TI - Absence of detrimental effects of fluoride exposure in diabetic rats. AB - This study is part of a comprehensive programme to investigate fluoride toxicity and the hypothesis that fluoride ingested by "medically compromised' animals will result in altered physiological function. Its objectives were to monitor fluoride retention, tissue fluoride concentrations and genetic variables in diabetic and control rats chronically exposed to fluoride, and to determine whether or not adverse effects occurred. Male, Zucker fatty diabetic rats and Zucker age-matched lean controls were fed a low-fluoride diet ( < 1.2 parts/10(6) F-) ad libitum and received 0, 5, 15 or 50 parts/10(6) fluoride in their drinking water for 3 or 6 months. Fluoride metabolic balance was determined for 4 days before the end of each study phase. Plasma and urine were analysed for biochemical markers of tissue function, and plasma, urine, faeces and tissues were analysed for fluoride. Bone marrow cells from animals killed after 6 months of treatment were examined for frequency of sister chromatid exchange, a marker of genetic damage. The diabetic rats consumed, excreted and retained significantly greater amounts of fluoride than the controls (p < 0.05). There were dose-related increases in fluoride excretion, retention and tissue concentrations in both classes of animals, which were significantly greater in the diabetic rats. In spite of greater amounts of fluoride in the tissues of diabetic animals, there was no evidence, under these experimental conditions, that any of the fluoride exposures tested caused measurable adverse effects on the physiological, biochemical or genetic variables that were monitored. PMID- 8712976 TI - Analysis of amelogenin mRNA during bovine tooth development. AB - The amelogenins are highly conserved enamel-matrix proteins that are essential for proper mineral formation. Transcriptionally active genes encoding the bovine amelogenin proteins reside on both the X and Y chromosomes. Comparison of relative levels of amelogenin mRNAs at various stages of development indicated that the X-chromosomal amelogenin message is at least six fold more abundant than the Y. Alternative splicing generates at least seven messages, five from the X primary transcript, and two from the Y. The two most abundant X-chromosomal amelogenin messages are approx. 850 and 450 nucleotides long, and nearly 10-fold more 850-nucleotide mRNA can be measured than 450 nucleotide, which has lost most of exon 6 by splicing. The predominant small message encodes leucine-rich amelogenin protein (LRAP), and amounts of LRAP message are relatively constant during development. However, the amelogenin message from which exon 3 has been spliced declines approximately 2.3-fold, when compared to total X chromosomal amelogenin transcripts, suggesting differential regulation of alternative splicing. In addition, a new exon was identified within genomic DNA, which was shown to be expressed by the use of reverse transcriptase-polymerase chain reaction, and the exons were renamed accordingly. This new exon-4 sequence is unusual in that it is not highly conserved between species. PMID- 8712978 TI - Induction of adenocarcinomas in the submandibular salivary glands of female Wistar rats treated with 7,12-dimethylbenz(a)anthracene. AB - In 12 male and 12 female Wistar rats, 7-9 weeks of age, a solution of 0.05 ml of 1% 7,12-dimethylbenz(a)anthracene (DMBA) dissolved in acetone was injected directly into the submandibular glands biweekly, after the gland had been injured. In some rats the carcinogen was injected 1 week after the injury. Each rat received six to seven injections. Swelling was observed in the submandibular gland region as early as 4 weeks after the last injection. The animals were killed 4-8 weeks after the last injection and glands with tumour tissues were processed for light microscopy. The control rats of the same age that received a corresponding amount of acetone only (three male, three female), carcinogen injection alone (three male, five female), and injury only (four male, four female) were killed at the same time. Histological examination revealed adenocarcinomas of the submandibular gland in 12/12 (100%) female rats, six of which were associated with fibrosarcomas. The adenocarcinomas basically consisted of ductal and glandular structures. Sometimes tubular, cystic, papillary-cystic and cribriform-like structures were also observed. Male rats mainly developed fibrosarcomas, although there was one squamous-cell carcinoma. The reasons for the sex difference are not known. One adenocarcinoma developed in the submandibular gland of a female rat (1/5) that received carcinogen injections alone. Thus direct injections of DMBA into the submandibular gland produce adenocarcinomas in female Wistar rats. PMID- 8712979 TI - Immunohistochemical localization of kallikrein in salivary glands of the Japanese monkey, Macaca fuscata. AB - The immunofluorescence staining using anti-human urinary kallikrein polyclonal antiserum was intense in the luminal rim of the striated duct cells, but decreased in the excretory duct cells and was weak in the intercalated duct cells. Western blot analysis demonstrated the monospecificity of the antiserum to glandular kallikrein in Macaca submandibular gland homogenate. The staining was observed to a similar degree as that in salivary glands with long and numerous striated ducts in the following order: the submandibular, the parotid and the sublingual gland. These findings suggest that kallikrein is secreted by exocytosis from the duct cells, mainly from the striated duct cells. PMID- 8712980 TI - Influence of oestrous cyclicity on certain metabolic alterations in the submandibular glands of female rats. AB - This report concerns changes brought about in the submandibular salivary gland during different stages of the oestrous cycle in the concentrations of total lipids, cholesterol and protein, together with an assessment of the activities of cyclic AMP-specific phosphodiesterase, ATPase, succinate dehydrogenase and an estimation of plasma glucose concentrations. As it is known that the ovarian steroids influence metabolic processes, it was thought desirable to study such metabolic alterations. During the transition from a dioestrous to oestrous stage, lipid accumulation occurred. However, during the metoestrous stage, catabolic tendencies predominated although there was some protein synthesis. PMID- 8712977 TI - Determination of enamel protein synthesized by recombined mouse molar tooth germs in organ culture. AB - Epithelial-mesenchymal interaction is a prerequisite for tooth morphogenesis. To study this interaction, inner enamel epithelium and dental papilla mesenchyme of molar tooth germs from a 16.5-day mouse embryo were dissociated enzymatically and cultured alone or after recombination. Characteristic matrix protein synthesized and secreted by recombined tooth germ was determined quantitatively by enzyme linked immunosorbent assay. The protein was detected in the culture of recombined tooth germ but not of dissociated enamel epithelium alone. The amount of enamel protein increased until 8 days in culture. Morphological differentiation of the recombined epithelial rudiment into ameloblasts and enamel protein production were confirmed. PMID- 8712981 TI - Complications of closed laparoscopy:is the Veress needle safe? PMID- 8712982 TI - Best practice parameters for management of rectal cancer: recommendations of the Colorectal Surgical Society of Australia. PMID- 8712983 TI - Carcinoembryonic antigen in the evaluation of therapy of primary and metastatic colorectal cancer. PMID- 8712984 TI - Colorectal resection in Victoria:a comparison of hospital based and individual audit. AB - BACKGROUND: The results of personal audit have not been tested against a hospital based audit previously and the results of two such audits of colorectal resection in the State of Victoria have provided this opportunity. In addition, data reflecting the results of colorectal resection across a range of hospitals and surgeons in the Victorian community have been obtained. METHODS: A total of 535 patients undergoing a colorectal resection, with an anastomosis performed, were studied in two serially conducted prospective audits arranged by the Standards Sub-Committee of the Victorian State Committee. One study was public hospital based and the second was based on voluntary reporting by individual surgeons. RESULTS: Similar results were obtained in each study, demonstrating the accuracy of individual reporting. The combined results (wound infection rate 12.3%, anastomotic leak rate 3.7% and mortality 4.5%) are compared to previously published data. CONCLUSIONS: In the State of Victoria the results of audit by individual surgeons performing colorectal resection were similar to the hospital based audit. The results obtained compare favourably with previously published data. PMID- 8712985 TI - A long-term follow-up of patients undergoing colectomy for chronic idiopathic constipation. AB - BACKGROUND: Chronic idiopathic constipation is a condition that mainly affects young women and is commonly associated with symptoms of abdominal pain and bloating. It has been proposed that patients with severe symptoms who are unresponsive to conservative measures can be managed by colonic resection. The aim of the present study was to assess the long-term outcome of such surgery on patients with a diagnosis of chronic idiopathic constipation. METHODS: Ninety-six patients (92 females, 4 males) underwent either a total colectomy and ileorectal anastomosis (n = 86) or subtotal colectomy and caecorectal anastomosis (n = 10) between 1986 and 1994. RESULTS: Postoperative mortality was 2.1%, 3.1% suffered from an anastomotic leak and 11.5% developed a pelvic abscess. Follow up was completed in 92.7% of patients at a mean of 5.0 +/- 2.3 years. Following surgery, symptomatic improvement was reported by 81.6% of patients. However, 51.2% still experienced difficulty with straining, 50.6% had some degree of anal incontinence, 55.2% continued to experience abdominal pains and 75.9% continued to be troubled by abdominal bloating. Reoperation was performed on 35.6% of patients (mainly for division of adhesions), and 9.2% of patients required an ileostomy. CONCLUSIONS: Colectomy is associated with relief of constipation in a majority of patients with chronic idiopathic constipation. However, it is associated with a considerable morbidity and is less effective in resolving symptoms of abdominal pain and bloating. PMID- 8712986 TI - Rib fractures in major trauma. AB - BACKGROUND: To determine the mortality, hospital and intensive care unit (ICU) stay of rib fractures in patients admitted to Victorian hospitals for more than 1 day. METHODS: All patients fitting the entry criteria for the Victorian Major Trauma Study with fractured ribs were identified between 1 March 1992 and 28 February 1993. Aetiology, age, sex, associated injury and outcome were analysed. RESULTS: Patients with rib fractures had a higher mortality and length of hospital stay, but this was not significantly different from other trauma. A significantly higher percentage of patients required ICU care for rib fractures (44%) compared with the total group with blunt injury (24%). The majority of rib fractures resulted from motor vehicle accidents 361/541 (67%). Injuries occurring on the street/highway resulting in rib fractures were more likely to be major; 62% had Injury Severity Score (ISS) > 15. Fractured ribs occurred more commonly with increased age. Mortality for patients with fractured ribs versus total trauma group was higher in elderly patients. Univariate analysis showed rib fractures were a positive predictor of death but when adjusted for ISS and age, rib fractures became a negative predictor. Rib fractures were not predictors for length of ICU or hospital stay. CONCLUSION: The sample of rib fractures collected in this study underestimates the overall incidence. For those patients admitted to hospital with identified rib fractures, there is a trend towards higher mortality and morbidity. However, this association is better predicted by ISS and age. PMID- 8712987 TI - Is penetrating injury on the increase in south-western Sydney? AB - BACKGROUND: Few Australian studies describe the epidemiology of penetrating trauma. This study describes the incidence and demographic features of penetrating injuries with emphasis on trends and severity analysis. METHODS: Case analysis was performed utilizing data from the Liverpool Hospital Trauma Registry (1989-94), NSW Department of Health Hospital Separations (1991-93), and the NSW Bureau of Crime Statistics (1991-93) with reference to the Liverpool and Fairfield Local Government Areas (LGA). RESULTS: The Trauma Registry revealed 251 of penetrating trauma. The median age was 26 years (interquartile range 21-33). Ninety-one per cent of the victims were male. Fourteen per cent of patients had an Injury Severity Score (ISS) > 15. Sixty-five per cent of cases were stab injuries and 20% gunshot injuries with the abdomen being the most commonly injured site. Twenty-one per cent of patients underwent laparotomy, 1.6% thoracotomy and 1.2% thoracotomy and laparatomy. There were 10 (4%) deaths. Trends in incidence of penetrating trauma and violent crime involving weapons were analysed. Static trends were observed for the annual incidence of penetrating trauma from the Liverpool Hospital Trauma Registry. Separations for penetrating trauma from Liverpool and Fairfield hospitals showed a slightly increasing trend. Violent crimes involving weapons in the Liverpool and Fairfield LGA showed a static trend. Nevertheless, separations for penetrating trauma and rates of violent crimes involving weapons were higher in south-western Sydney than metropolitan Sydney or NSW. Eight per cent of the LGA population are Vietnamese but this study identified 16% of victims as being Vietnamese. CONCLUSIONS: This study found no significant increase in penetrating trauma or violent crime predisposing to penetrating injury in south-western Sydney. PMID- 8712988 TI - Bleeding giant gastric ulcer. AB - BACKGROUND: A consecutive series of 269 cases of bleeding gastric ulcer were studied prospectively from 1979 to 1993 inclusive. METHOD: Fifty-five (21%) had a giant gastric ulcer with a diameter of 3 cm or more. These cases were compared with those with ulcers less than 3 cm in diameter in terms of clinical details, prognostic factors, urgent operation and outcome. RESULTS: Death occurred in 13 cases (5%), urgent surgery was performed in 75 cases (29%) and there were 11 postoperative deaths (15%) within a month of surgery. The patient details in the two groups matched in terms of age, sex distribution, ulcer history, previous complication and recent ingestion of analgesics. Clinical comparison showed that giant ulcer had a poorer prognosis with a higher mortality (10 vs 3%, P < 0.01), urgent surgery rate (65 vs 12%, P < 0.01) and operative mortality (23 vs 11%, difference not significant). Study of risk factors in patients with giant ulcer revealed significantly more with concurrent illness, shock, anaemia and endoscopic stigmata of recent haemorrhage. CONCLUSION: More severe bleeding and poorer general condition in the giant ulcer group stresses the importance of early diagnosis and accurate resuscitation in these patients. Survival depends on optimal condition and prompt and timely surgery. PMID- 8712989 TI - Does the colon adopt small bowel features in a small bowel environment? AB - BACKGROUND: Isoperistaltic colon interposition is one of several surgical options available in the treatment of short bowel syndrome. It functions primarily by slowing the rapid intestinal transit that follows small bowel resection (SBR). Previous studies have interposed distal colonic segments and although there is evidence of adaptation by the interposed colon it is not known whether these segments develop histologic or functional small bowel characteristics. METHODS: In this study evidence was sought of metaplasia in the interposed segments after 80% SBR and, because proximal colon is a midgut derivative, the differences between proximal and distal colonic segments were examined. RESULTS: There was no qualitative histological evidence of metaplasia by the interposed segments. There was a significant increase in crypt depth, mucosal thickness and maltase concentration of the interposed segments compared with the nontransposed colon of the controls. The maltase concentations were increased to the extent that they were not significantly different from the concentration present in normal ileum. CONCLUSIONS: Although there was no gross evidence of small bowel intestinalization by the interposed segments, there was evidence of adaptation which was not merely an extension of the adaptive process seen after SBR alone. These changes resulted in the colon taking on some small bowel features. There was no significant difference between proximal vs distal interposed segments. PMID- 8712990 TI - Preclinical trial of a bioartificial liver support system in a porcine fulminant hepatic failure model. AB - BACKGROUND: This study describes the pre-clinical trials of an extracorporeal bioartificial liver support system (BALSS). It includes the biochemical changes which occur in the plasma and blood of pigs with devascularized livers when the plasma is treated in a BALSS, and the testing of the system for presence or absence of infective agents, pyrogens and for toxicity. METHODS: Hepatic cells were prepared from littermate juvenile white landrace pigs with a double-step collagenase digest technique. The cell preparations were incubated with collagen coated dextran microspheres (CDM) for 3 h and the medium was tested to determine cellular metabolic activity. Incubation continued for a further 20 h during which the hepatic cells attach to the CDM. The CDM-attached cells were inoculated into a hollow fibre bioreactor which was part of an extracorporeal liver support system. RESULTS: Hepatic cell content of the bioreactor was 6 x 10(9) +/- 3 x 10(8) cells, equivalent to those present in half a pig's liver. The system was tested in a controlled trial with the plasma of pigs with fulminant hepatic failure (FHF) due to devascularized livers. When plasma from FHF pigs was circulated through the device there was significantly less of an increase in the accumulation of ammonia, lactate and most amino acids when hepatic cells were included in the circuit compared with those in control experiments when they were excluded. Similar changes occurred in procine blood. There were few infections diagnosed and an absence of pyrogens, endotoxins and toxicity in the bioreactor contents or in the terminating reservoir or animal blood samples. CONCLUSIONS: We believe that the results, demonstrating function of the porcine hepatic cells in the circuit, together with low risks, justify a clinical trial of use of the BALSS in Australia. PMID- 8712991 TI - Minimally invasive neurosurgery. AB - BACKGROUND: The philosophy and practice of minimally invasive surgery have fundamentally altered the practice of general and gynaecological surgery, and are currently transforming the practice of neurosurgery. The goal of minimally invasive surgery is to reduce tissue disruption and thus morbidity. This is a review of the development, applications, and benefits of minimally invasive neurosurgery, and its wider surgical implications. METHODS: A synthesis of the literature is presented. RESULTS: Interactive computer imaging with frame-based and frameless stereotaxy, ultrasonographic and endoscopic techniques are increasingly being used independently and in combination in order to execute minimally invasive approaches and to navigate a safe path within the cranium or spine. Interaction using an interlinking stereotactic instrument or robot between the surgeon, a graphic interface (such as a computer workstation) and the patient, enables precise planning and execution of surgery with exact correspondence of imaging data and the living pathology. The direct influence of virtual reality, cybernetics, robotics, and telepresence will further revolutionize the practice of neurosurgery and will impact increasingly on other surgical disciplines. Safety and precision are further enhanced by intra operative physiological monitoring techniques. Interventional neuroradiology and stereotactic radiosurgery add further dimensions to the minimally invasive approach and may not only diminish the need for open surgery, but in selected cases obviate the need altogether. CONCLUSIONS: Minimally invasive neurosurgery is a major force in contemporary neurosurgery and many of the current neurosurgical applications will have far-reaching effects on the practice of surgery in general. PMID- 8712992 TI - Recent developments in upper gastrointestinal surgery. PMID- 8712993 TI - Controlled duodenostomy for difficult duodenal stump. PMID- 8712994 TI - Bone allograft banking in South Australia. PMID- 8712995 TI - Avulsion of the ischial apophysis. AB - Avulsion of the ischial apophysis is a rare injury. We report the successful treatment of an avulsion of the ischial apophysis by open reduction and internal fixation. PMID- 8712996 TI - Pseudomembranous panenteritis and septicaemia in a patient with ulcerative colitis. AB - Pseudomembranous colitis may occur in the immunocompromised chronic colitic patient without a prior history of antibiotic use. The entire gastrointestinal tract can be involved and the presentation can be that of a severe systemic infection that warrants prompt recognition and aggressive management. PMID- 8712997 TI - Left scrotal abscess complicating laparoscopic appendicectomy. PMID- 8712999 TI - Metachronous colorectal and biliary carcinoma: the aetiological implications of k ras oncogenes. PMID- 8712998 TI - Idiopathic chondrolysis of the hip. AB - A case of idiopathic chondrolysis of the hip is described in a 12 year old female who has the characteristics of arthroophthalmopathy (Stickler's syndrome). This common connective tissue dysplasia may go unrecognized and is possibly an aetiological factor in the development of this disease process. A review of the literature is presented. PMID- 8713000 TI - Neuropeptides, nerve growth factor and eczema. AB - Considerable evidence indicates that peripheral nerve fibres play an active regulatory role during eczematous reactions. Many basic features of the inflammatory process are potentially subject to neuropeptidergic modulation, such as vascular changes, cellular trafficking activation and trophism in vivo and in vitro investigations suggest that neuropeptides are part of the complex network of mediators that initiate and maintain the eczematous process in the skin. In general, peptidergic fibres seem to exert a global protective role, but with specific and selective effects in the different phases. Evidence that the inflammatory process induces bio-chemical changes at a neuronal level has also been obtained. The possible mediators of the neuronal recruitment in the course of the eczematous reaction are still unknown. Nevertheless, nerve growth factor, which is locally increased during inflammatory processes and may induce peptidergic alterations, should be considered as a potential candidate molecule. PMID- 8713001 TI - The Australasian College of Dermatologists and Japanese Dermatological Association joint meeting. Cairns, Australia, 3-4 November 1995. Proceedings. PMID- 8713002 TI - Compliance problems in paediatric atopic eczema. AB - Compliance problems are a common cause of apparent treatment failure in atopic eczema. The results of a survey of parents with children attending dermatologists for atopic eczema is presented. Major reasons for failure to comply include a poor understanding of the chronic nature of the disease, a fear of topical corticosteroids, the belief that atopic eczema is due to an allergy, and the time and cost of treatment. The explanation that treatment is a control and not a cure is also important in the parent's perception of treatment results. Suggested solutions to some of these points are discussed. PMID- 8713003 TI - Viral exanthems in childhood. AB - Viral exanthems can be classified as erythematous vesicular and papular. The majority are erythematous with the most common viral causes being non polio enteroviruses, respiratory viruses, acute. Epstein-Barr virus, human herpes viruses 6 and 7, and parvovirus B-19. Measles, rubella, mumps, acute GMV, hepatitis viruses. HIV seroconversion, Ross River and Barmah Forest viruses are less commonly seen. The differential diagnosis includes drug eruption, erythematous bacterial exanthems and Kawasaki's syndrome. PMID- 8713004 TI - Pathological twists in association with infectious agents and infestations of the skin. AB - The capacity of micro-organisms and infestations to induce flare or mimic a wide variety of dermatological disorders has long been appreciated. The development of new techniques such as in situ hybridization and the polymerase chain reaction has permitted a re-examination of old concepts such as the tuberculids and bacterids. In addition new discoveries have highlighted the biological potential of micro-organisms to stimulate or depress the immune system through potent biological active products and has led to new insights in the mechanisms through which organisms may produce skin changes. Histological simulants of dermatological disease may also be induced by micro-organisms and these may need to be distinguished by clinical correlation. PMID- 8713005 TI - Human T lymphotropic virus-1 infection. AB - Human T lymphotropic virus-1 (HTLV-1) is a retrovirus which infects T lymphocytes (CD4+) to cause adult T cell leukaemia/lymphoma (ATL), tropical spastic para paresis and several other HTLV-1 associated disorders. ATL has been reported worldwide but areas of high incidence include Japan (particularly the south west), Central and South America, northern Iran, West and Central Africa and Melanesia. In the general Japanese population, HTLV-1 carriage is 0.1% but this can be as high as 50% in endemic areas. Six per 10000 carriers are estimated to progress to ATL each year. The three major routes of infection are mother to baby through breast-feeding, sexual intercourse and blood transfusion. There is a lengthy latency period of up to 40 years before the development of ATL. Up to 50% of ATL patients present with a cutaneous eruption. Diagnosis is established by the detection in lymphocytes of monoclonal integration of HTLV-1 proviral DNA. Even with aggressive treatment, ATL patients generally have a poor prognosis. PMID- 8713006 TI - Marine stingers in far north Queensland. AB - Two box jellyfish in particular cause problems in tropical Queensland waters. Chironex fleckeri inhabit calm waters close to the shore between November and May. The venom includes three major components: haemolytic dermatonecrotic and myocardial. The dermatonecrotic toxin causes a ladder pattern of whiplash lesions to the skin which ulcerate become necrotic and heal very slowly over months: Neuromuscular paralysis and cardiovascular collapse may be fatal within minutes of envenomation. Emergency treatment comprises inactivation of stinging capsules by vinegar removal of tentacles analgesia, cardiopulmonary resuscitation and the administration of the specific antivenom. Carukia barnesi ('Irukandji') are found in both coastal and open waters. A patch of erythema with papules at the sting site is characteristically followed 30 min later by the onset of a catecholamine mediated syndrome. Headache and severe abdominal and back pain are usual and may be followed by hypertension, tachyarrhythmias and cardiogenic shock. PMID- 8713007 TI - Psoriasis: changing clinical patterns. AB - Although psoriasis has been recognized at least since Biblical times new forms, associations and influences continue to be described in the twentieth century. New forms include the rupioid erythema annulare centrifugum-like and follicular patterns. Associations with vitiligo bullous pemphigoid and lupus erythematosus have been recently described. Endoscopic surgery has increased para umbilical psoriasis while Sun Smart campaign have reduced photo-aggravated psoriasis. Infections such as paediatric perianal streptococcal cellulitis and drugs including angiotensin converting enzyme inhibitors and cytokines exacerbate psoriasis. PMID- 8713008 TI - A quiet revolution: a surgical approach to skin cancers of the forehead. AB - Dermatologists are performing increasing numbers of procedures for skin cancers with specialized training in a number of treatment modalities. As an example, repairs of surgical defects of the forehead are presented including rotation flaps, O to T closures and Burow's triangle flap. PMID- 8713010 TI - Skin phototyping in Asian Australians. AB - Skin phototype was assessed in 257 Asian Australians by self-reporting questionnaire. Minimal erythema dose, minimal melanogenic dose and minimal immediate pigment darkening dose were measured in a subgroup of 50 subjects. About 15% of Asian Australians in this study report that they have skin type I or II. Phototesting confirms that there is a UV-sensitive group and a wide spectrum of UV-sensitivity in this population. Whether Fitzpatrick's skin typing system adequately identifies this UV-sensitive group needs assessment by a larger study. The relationship between burning tendency and tanning capacity in Asians may differ from Caucasians. PMID- 8713009 TI - Treatment of B cell lymphoma of the skin. AB - Ten patients with B cell lymphoma of the skin have been treated in a 5 year period at the Keio University Hospital, Tokyo. Most present with an asymptomatic solitary nodule or multiple tumours of the skin but limitation to the skin is rare. Histopathology showed medium to large lymphoid cells in the dermis and subcutis. Radiotherapy was given for cutaneous lesions in conjunction with chemotherapy if there were metastases. PMID- 8713011 TI - Putative linkage between HLA class I polymorphism and the susceptibility to malignant melanoma. AB - Human leukocyte antigen (HLA) frequencies in 67 melanoma patients were compared to 120 controls in Japan. The frequency of HLA-B13 was increased in melanoma patients compared to controls and was particularly common in nodular melanoma HLA B61 was decreased in melanoma patients. Almost 50% of acral lentiginous melanoma patients expressed HLA-B51 compared to 0% of nodular melanomas. PMID- 8713012 TI - Psoriasis: mechanisms and entry points for possible therapeutic interventions. AB - Psoriasis is a heterogeneous disorder not only in regard to its variability in clinical expression and age at onset, but also in the degree to which genetic and immunological factors such as T cells and cytokines contribute. A Psoriasis Treatment Index' is proposed according to clinical severity and hence suggested treatment regimens. Recent and new treatments include FK 506, cyclosporin, a fusion protein of human interleukin 2 with fragments of diphtheria toxin, topical tumour necrosis factor inhibitors, topical retinoids, T cell receptor peptide vaccines and intercellular adhesion molecule 1 (ICAM-1) antisense oligonucleotides. PMID- 8713013 TI - Rheological analysis of peripheral blood neutrophils in a patient with cutaneous and arthropathic psoriasis accompanying hypopyon-iridocyclitis. AB - A 32 year old Japanese male has had several synchronized attacks of psoriatic arthritis and hypopyon-iridocyclitis. During exacerbations, he has a leucocytosis with neutrophilia and elevated C-reactive protein. Assessment for adhesiveness, aggregation and deformation of peripheral neutrophils revealed strikingly high rheological activity whereas during periods of remission the activity was only slightly elevated above normal. PMID- 8713014 TI - Two cases of protothecosis in Nagoya, Japan. AB - Two cases of protothecosis caused by Prototheca wickerhamii have been reported from Nagoya in a 12 year period in both cases the infection presented on the cheeks of otherwise healthy women. Biopsies showed numerous PAS positive staining organisms with the distinctive mulberry like endosporulation in the dermis. Prototheca wickerhamii was identified on sugar assimilation tests of colonies isolated from tissue on Sabouraud agar. Case 1 responded to 11 months of oral ketoconazole therapy. Case 2 might not respond to itraconazole. The source of the infections has not been identified. PMID- 8713015 TI - Time-course analyses of exercise-induced lesions in a patient with urticarial vasculitis. AB - Using immunoperoxidase techniques, we examined the histological changes of exercise-induced lesions of urticarial vasculitis over a 24 h period. At 3 h, endothelial swelling and an eosinophilic infiltrate was seen. Neutrophils were increased in number by 10 h. Typical leucocytoclastic vasculitis with red blood cell extravasation developed by 24 h. Maximal deposition of eosinophil peroxidase was at 10 h compared to significant neutrophil elastase deposits at 24 h. This demonstrates the importance of the eosinophil in the development of the lesions of urticarial vasculitis. PMID- 8713016 TI - Advice about management of skin conditions in the community: who are the providers? AB - A survey was undertaken in the central Victorian city of Maryborough to investigate the use of pharmacies and general practitioners as sources of advice about skin problems. In a 2 week period consumers purchasing either prescription or over the counter (OTC) products for skin, hair and nails in pharmacies were approached for interview. Simultaneously, each of the city's general practitioners (GP) completed questionnaires on all patients presenting with conditions of skin, hair and nails. Seventy per cent of the 315 consumers interviewed were purchasing OTC products and 50% prescription items. Of the OTC products, 42% were originally recommended by pharmacy staff and 18% by doctors. Over one-third of consumers buying OTC products described symptoms to pharmacy staff and in about half of the cases it was to the pharmacy assistant. One-third of people describing symptoms had already seen a doctor. Problems most frequently reported were dermatitis, skin dryness, acne and tinea. General practitioners recorded 265 conditions (of which 54% were new) in patients. The most common condition treated was solar keratosis, and GP wrote a prescription for 45% and recommended OTC products for 7% of conditions. People in Maryborough are seeking advice for their skin conditions from a variety of sources including GP, pharmacies and others. PMID- 8713017 TI - Tissue factor and thrombomodulin expression on keratinocytes as coagulation/anti coagulation cofactor and differentiation marker. AB - Immunohistochemistry was used to investigate the expression of tissue factor (TF) as a coagulation factor and thrombomodulin (TM) as an anticoagulation cofactor in the epidermis. TM was expressed on Malpighian layer keratinocytes while TF was located on the supra-Malpighian layer. Epidermal shave extracts were biologically active for both factors. Keratinocytes cultured in high calcium, but not low calcium media, expressed both TF and TM. In pemphigus vulgaris pemphigus foliaceus and bullous pemphigoid, TF expression was increased on keratinocytes shielding the blister compared to the down-regulated TM expression by keratinocytes around the blister. PMID- 8713018 TI - The frequency and nature of skin conditions seen in a private dermatology practice in central Victoria, 1991-95. AB - A dermatology diagnostic database was established in order to determine the frequency and nature of skin conditions in a private dermatological practice in Central Victoria, Australia. Between August 1991 and June 1995, a total of 3346 dermatological diagnoses were recorded for 3000 new patient presentations. More women (58.9%) were seen than men (41.1%). Dermatitis and tumours of the skin accounted for 50% of new consultation(s), the highest attendance being for solar keratoses (11.9%). The frequency of patients seen was comparable to the previous 1970s survey of private dermatological practice in Australia, with the exception of melanocytic naevi, which was seen with twice the frequency compared to previously. The proportion of patients presenting with dermatitis was higher in the colder seasons of the year (winter and spring compared with summer and autumn). The proportion of people seen with melanocytic naevi was higher in summer and autumn compared with winter and spring. These results illustrate the usefulness of recording clinical patient data in practice as a way of recording patterns of referral as well as having a role in determining causation of disease. PMID- 8713019 TI - Immune response to Streptococcus pyogenes and the susceptibility to psoriasis. AB - Monoclonal antibodies directed against type 12 Group A streptococcal cell wall antigens cross-react with nuclei and cytoplasm of cells from skin and synovium from controls, uninvolved skin of psoriatics and psoriatic plaques. Patients with psoriasis had high serum titres of antibody against the M12 (C-region) streptococcal antigen compared to controls. An abnormal immune response directed against a "self' antigen after initiation by Group A streptococcal infection may play an important role in the exacerbation or development of psoriasis. PMID- 8713020 TI - Atopic eczema of adult type in Japan. AB - Atopic eczema of adult type presents with the classical features of eczema in combination with a red face subject to oozing attacks rippling hyperpigmentation of the neck and an exudative erythema of the trunk and limbs. The red face and hyperpigmentation appear to follow topical corticosteroid use while the exudative erythema is postulated to be due to late phase reaction of IgE. Treatment involves the avoidance of exacerbating factors together with topical emollients. Topical corticosteroids are ineffective and appear to exacerbate the red face, which causes a significant social problem, affecting the patient's quality of life. New therapies promoted through the media have caused confusion. PMID- 8713021 TI - Cyclophosphamide (Cytoxan)-induced hematologic alterations in specific-pathogen free chickens. AB - Cyclophosphamide (Cytoxan) was given at 75 mg/kg body weight via daily intramuscular injections for 4 days to 3-week-old specific-pathogen-free (SPF) chickens in an attempt to determine if heteropenia could be induced in chickens. Control birds were given a like quantity of phosphate-buffered saline, the diluent for Cytoxan. Peripheral blood heterophil numbers were determined and monitored by total leukocyte and differential cell counts. Birds were grouped in pairs on day 0 based on total leukocyte count. The number of heterophils each bird had on day 0 served as a baseline heterophil count for that bird. Thereafter heterophil numbers were determined on the last day of drug treatment and every other day until blood heterophil numbers were 20% of that bird's baseline heterophil count (heteropenia). The effects of Cytoxan on trachea, lung, liver, kidney, bursa of Fabricius, bone marrow, spleen, and thymus were determined by microscopic examination of those tissues collected the day following heteropenia. Cytoxan had no effect on trachea, lung, liver, kidney, and thymus. Bursa of Fabricius and spleen had decreased amounts of lymphoid aggregates. Bone marrow of Cytoxan-treated chickens was hypocellular. The study was then repeated to determine the reversibility of Cytoxan-induced heteropenia. Cytoxan-treated birds were allowed to recover until blood heterophil numbers equaled or exceeded those of control birds. Cytoxan, through bone marrow suppression, induced a reversible heteropenia that developed between treatment days 10 and 12. In addition, Cytoxan induced a reversible lymphocytopenia between days 4 and 10 and a regenerative anemia between days 8 and 10. The ability to produce heteropenia in SPF chickens will allow the use of a heteropenic model for further study of the heterophil's contribution to the inflammatory response. PMID- 8713022 TI - Primary cell culture of turkey intestinal epithelial cells. AB - Primary cell culture has been widely used in various types of studies and proven useful for the isolation and identification of avian pathogens. Difficulties in growing intestinal epithelial cells in vitro have limited their use for such studies. In the present study, a co-culture system was developed for the primary culture of intestinal epithelial cells. A monolayer obtained from 14-to- 16-day old turkey embryo intestinal fibroblasts was used as a feeder layer. Feeder layers from turkey embryo fibroblasts and from a continuous cell line (mouse 3T3 fibroblasts) were also employed but were not as successful. The intestinal epithelial cells were isolated by dissociation from the intestinal tracts of 1 day-old turkey poults and grown on the feeder layers. Growth and maintenance media were supplemented with various components, including fetal calf serum, chicken serum, hormones, and other growth factors. The epithelial cells grown on feeder layers from the intestinal fibroblasts allowed the intestinal epithelial cells to be maintained in vitro for periods of 7 to 10 days. This technique may prove useful for various applications, including isolation of enteropathogens, and for basic studies of the intestinal tract concerning such subjects as physiology, immunology, and toxicology. PMID- 8713023 TI - Apoptosis in cell cultures induced by infectious bursal disease virus following in vitro infection. AB - Chicken embryo fibroblasts (CEFs) and Vero cells infected with infectious bursal disease virus (IBDV) exhibited the biochemical feature of apoptosis. Agarose gel electrophoresis of DNA extracted from IBDV-infected cells revealed the characteristic laddering pattern of DNA fragmentation, which was more intense in infected CEFs than in Vero cells. The appearance of apoptotic nucleosomal DNA fragments in IBDV-infected CEFs was independent of virus replication and occurred at an early stage following in vitro infection. PMID- 8713024 TI - Avian infectious bronchitis: viral persistence in the harderian gland and histological changes after eyedrop vaccination. AB - The histological changes in the harderian gland (HG) induced by the attenuated H 120 infectious bronchitis virus (IBV) vaccine strain and the persistence of this virus in the stroma of the gland was evaluated in chickens after eyedrop vaccination. Virus replication induced an increase in IBV-specific enzyme-linked immunosorbent assay antibody levels from marginal levels at vaccination (26 days of age) to significantly higher levels 10 days after exposure. IBV antigen was detected in the HG by both immunofluorescence using a monoclonal antibody and virus reisolation in embryonated chickens eggs until day 14 postvaccination. Lymphocytic, heterophilic, erythrocytic, and plasma cell infiltration as well as epithelial cell integrity in collecting tubules and acini were evaluated in the HG throughout the experimental period. IBV vaccination with the attenuated vaccine strain H-120 resulted in partial damage to the HG, as demonstrated by both the presence of plasma cells showing Russell bodies and by tubule epithelial cell exfoliation that occurs simultaneously with the presence of detectable IBV. The increase in plasma cell number and the enlargement of lymphoid foci appear to be expressions of the immunocompetence of this paraocular gland. PMID- 8713025 TI - Causes of morbidity in bustards in the United Arab Emirates. AB - The findings of 1746 clinical examinations of 594 bustards of six different species are presented, and the differences of causes of morbidity between imported adult, captive adult, and captive juvenile bustards are discussed. Wing tip trauma, keel trauma, and other soft tissue-related traumatic injuries were the most commonly diagnosed clinical conditions in captive adult bustards, accounting for 50.7% of the total findings in this category. Soft tissue-related related traumatic injuries accounted for 26.3% and 5.3% of the findings of imported adult and captive juvenile bustards, respectively. Musculoskeletal disorders were the most commonly diagnosed clinical conditions in captive juvenile bustards, accounting for 61.3% of the total findings in this category. Nutritional bone disease was the single most important musculoskeletal disorder in captive juvenile bustards, accounting for 24% of total findings. Musculoskeletal disorders accounted for 22.9% and 4.9% of findings in captive adult and imported adult bustards, respectively. Parasitic observations accounted for 24.7% of the total findings in imported adult bustards. Infectious viral diseases were almost exclusively confined to the imported adult bustards, making up 20.1% of findings in this group. Opthalmologic conditions accounted for 15.1% of findings in imported adult bustards. PMID- 8713026 TI - Comparison of enzyme-linked immunosorbent assay and hemagglutination-inhibition for detection of antibody to Mycoplasma gallisepticum in commercial broiler, fair and exhibition, and experimentally infected birds. AB - Hemagglutination-inhibition (HI) assay and a new affinity-purified enzyme-linked immunosorbent assay (ELISA) for detection of antibody to Mycoplasma gallisepticum (MG) compared for use as confirmatory tests for the National Poultry Improvement Plan program. Samples from three different poultry populations with different prevalences of MG infection were studied: commercial broiler breeder birds (low prevalence of infection), fair and exhibition birds (moderate prevalence of infection), and experimentally infected birds (high prevalence of infection). Western immunoblots were used to confirm infection status in samples that had discrepancies between HI and ELISA results. The prevalence of infection in commercial broiler birds and in exhibition and fair birds in Florida was determined. Samples from culture-positive Mycoplasma synoviae (MS) flocks also were tested and compared for potential cross-reactions. The prevalence of MG infection was very low (< 1%) in the commercial population, with no significant difference between the HI and ELISA test results (P = 0.3157). The prevalence of MG infection in the fair and exhibition birds tested was approximately 40%, and ELISA was more accurate than HI for confirmation of MG infection in this population (P = 0.036). In birds experimentally infected with MG, there was no significant difference between HI and ELISA results (P = 0.6542). Of the 195 sera collected from flocks confirmed positive for MS by culture, 15% cross-reacted with the MG serum plate agglutination (SPA) test. There were no cross-reactions observed with either the MG ELISA or HI. There was a positive correlation of HI titers to ELISA values (R = 0.621, P < 0.01). Results from this study showed there were no differences between ELISA and HI as confirmatory tests in populations with a low prevalence of MG infection. However, ELISA was superior to HI in a population with moderate levels of MG infection. PMID- 8713027 TI - Comparison of the genomic short regions of a vaccine strain (SA-2) and a virulent strain (CSW-1) of infectious laryngotracheitis virus (Gallid herpesvirus 1). AB - Restriction enzyme linkage maps were produced for the genomic short region of the virulent infectious laryngotracheitis virus (CSW-1 strain). After comparison with the equivalent restriction enzyme linkage maps for the infectious laryngotracheitis virus SA-2 strain (a vaccine strain), it was determined that the maps for the short regions of the two strains were identical, apart from a single section in each of the inverted terminal repeats. Each inverted terminal repeat of the SA-2 strain was discovered to contain 467 base pairs more DNA than the CSW-1 strain's inverted terminal repeats. This extra DNA was more precisely mapped entirely within the EcoRI fragments D and d of SA-2, which were found to form part of the SmaI fragments U and P of SA-2 and Q and b of SA-2 and to contain one SmaI restriction enzyme site. PMID- 8713028 TI - Characterization of the avian polyomavirus-associated glomerulopathy of nestling parrots. AB - The glomerulopathy occurring in nestling nonbudgerigar parrots with avian polyomavirus (APV) disease was examined in 10 parrots. Glomerular lesions were characterized by the presence of dense, periodic acid-Schiff (PAS)-positive material that expanded the mesangium and that narrowed and at times occluded capillary lumina. PAS-staining was found to be more sensitive than hematoxylin and eosin in the detection of the lesions. Ultrastructurally, finely granular electron-dense material was found in massive intracapillary and mesangial condensates. Capillary endothelial cells exhibited changes consistent with cellular swelling. No evidence of chronic glomerular changes was observed. Immunofluorescent staining demonstrated that the PAS-positive, electron-dense condensates were complexes of IgG, avian polyomavirus antigen, and, in one bird, IgM. Viral DNA was detected in the serum of all six birds examined. Anti-APV antibodies were also present in all five serum samples examined. These findings suggested that the pathogenesis of this acute immune complex glomerulopathy and other APV-associated lesions depends on the presence of an appropriate ratio of circulating virus and anti-APV antibody. PMID- 8713029 TI - Characterization of Bangor virus proteins by using monoclonal antibodies. AB - A new virus was isolated from a finch in quarantine in Northern Ireland in 1973. The virus had the morphological characteristics of a paramyxovirus, and was named Bangor virus (BaV). In order to identify the structural proteins of BaV and to investigate the biological characterization of the virus, 28 monoclonal antibodies (mAbs) directed against BaV were prepared. Eight of these mAbs reacted with the nucleocapsid protein (NP), 10 with hemagglutinin-neuraminidase (HN) protein, and 10 with fusion (F) protein. With the aid of these mAbs, the structural proteins of BaV were determined, namely, p52, gp74, gp63, and gp51 were identified as the NP, HN, F0, and F1 proteins, respectively. The biological activities of the mAbs directed against the envelope glycoproteins of BaV were examined. Intriguingly, it was found in the neutralization assay that four mAbs directed against the HN protein of BaV can enhance the fusion of HeLa cells infected with BaV, showing the presence of a potential third function of the HN protein that affects the fusion activity of the F protein. Furthermore, all of the anti-F protein mAbs showed neutralizing activity. PMID- 8713030 TI - Experimental reproduction of severe hypoglycemia and spiking mortality syndrome using field-derived and embryo-passaged preparations. AB - The clinical signs, enteritis, weight depression, and hypoglycemia of spiking mortality syndrome were experimentally reproduced in broiler breeders and broiler chicks. Inocula included 1) virus-like particles from intestines of chicks with spiking mortality syndrome that had been banded in a discontinuous Renograffin gradient, 2) homogenized darkling beetles collected from litter of farms where spiking mortality syndrome had occurred repeatedly, and 3) homogenized embryos which had been inoculated with the Renograffin-banded material. Arkansas variant infectious bronchitis virus and arenavirus-like particles were identified in the inocula. Serology on samples from surviving chicks suggested the presence of an avian encephalomyelitis virus in one of the inocula. One-day-old (n = 172) and 2.5-day-old (n = 30) chicks were inoculated orally, and some were also injected intraperitoneally or subcutaneously, with 0.5 ml of the inocula. Twelve to fourteen days postinoculation, chicks were fasted for 4-6 hours, then briefly stressed with a cool water spray. Within 1.5 hours, inoculated chicks began dying with severe hypoglycemia and clinical signs of spiking mortality syndrome. Body weights were significantly depressed. Uninoculated controls (n = 130) from the same hatches, also fasted and stressed, were unaffected clinically and were not hypoglycemic. One group (n = 52) of inoculated chicks exposed to a controlled lighting program was unaffected clinically, had significantly higher mean plasma glucose levels, and had significantly less body weight depression than chicks exposed to continuous lighting. We concluded that exposure to controlled amounts of light/darkness can ameliorate much of the hypoglycemia, mortality, and runting stunting associated with spiking mortality syndrome of chickens. The significance of the viruses and virus-like particles detected in the inocula is currently under investigation. PMID- 8713031 TI - Efficacy of a recombinant fowl pox-based Newcastle disease virus vaccine candidate against velogenic and respiratory challenge. AB - A fowl pox-based recombinant virus TROVAC-NDV (vFP96.5) was developed expressing the fusion and hemagglutinin-neuraminidase glycoproteins from a velogenic strain of Newcastle disease virus (NDV). Studies in specific-pathogen-free birds indicated that inoculation of a single dose of the recombinant led to the induction of significant levels of hemagglutination-inhibiting antibody that were maintained to 8 wk postinoculation. Further, the recombinant induced protective immunity against a combined intramuscular velogenic NDV challenge and respiratory NDV challenge. In commercial broiler chickens that were inoculated in the presence of maternally derived NDV immunity, the level of the NDV-specific humoral response was dampened, but significant levels of protection against both a lethal intramuscular NDV challenge and a fowl poxvirus challenge were obtained. PMID- 8713032 TI - Runting of ducklings associated with Cochlosoma anatis infection. AB - Ducklings, especially common goldeneye (Bucephala clangula), experiencing poor weight gain and delayed development were reported from a waterfowl park during June and July of 1990. Runting was first noticed between days 5 and 10 post-hatch in the "brooder" building, and although ducklings appeared active and were feeding, they developed at a slower rate than other members of their clutch. Many ultimately died of emaciation and gram-negative bacterial septicemia. Necropsies of affected ducklings revealed large numbers of the intestinal flagellate Cochlosoma anatis, in both the small and the large intestine; however, autolysis impaired histological interpretation in many cases. Inoculation of 1-day-old Muscovy ducklings with feces containing large numbers of C. anatis resulted in poor weight gain and delayed tail-feather development compared with uninoculated control ducklings. Weight gain improved after treatment with metronidazole. Cochlosoma anatis is associated with the runting syndrome observed in ducklings at the waterfowl park. PMID- 8713033 TI - Efficacy of Salmonella enteritidis (SE)-immune lymphokines from chickens and turkeys on SE liver invasion in one-day-old chicks and turkey poults. AB - We have shown previously that increased resistance to Salmonella enteritidis (SE) organ infectivity in 1-day-old chicks was conferred by the immunoprophylactic administration of SE-immune lymphokines (SEILK). These lymphokines have been found to be present in the cell culture media of concanavalin A-stimulated splenic lymphocytes obtained from SE-immunized chickens. In the present study we evaluated whether turkeys also produced SEILK and whether these lymphokines could protect 1-day-old chicks and turkey poults against SE liver invasion. In addition, we tested the ability of our original chicken SEILK to reduce SE liver invasion in turkey poults. Day-of-hatch chicks and turkey poults were injected intraperitoneally with immune lymphokines of either chicken or turkey origin. One hour later the birds were challenged per os with SE, and 20 hours later their livers were examined by bacteriological methods for the presence of SE. We found that SEILK induced from the splenic lymphocytes of SE-immunized turkeys reduced SE liver invasion in both chicks and turkey poults. Conversely, we also determined that SEILK produced by chicken splenic lymphocytes conferred protection against invasion by SE in turkey poults. This research is the first report of the production of SEILK in turkeys and also the first report on the cross-species activity of these effector molecules in chickens and turkeys. PMID- 8713034 TI - Tween 80-solubilized Newcastle disease virus prepared as a water-in-oil-in-water vaccine. AB - The Newcastle disease virus (NDV) was solubilized with 10% w/v Tween 80 and inactivated with 0.05% v/v formalin. The average molecular mass of the released antigenic subunits was 307 kD. The tested vaccine was prepared in the form of a water-in-oil-in-water emulsion (WOWE) vaccine. The oil-to-aqueous ratio was 1:2. The solubilized NDV was administered alone or built in a tetravalent WOWE vaccine. A dose of the monovalent vaccine containing an equivalent of 44.7 microliters of detergent-treated NDV-allantoic fluid (NDV-AF) was sufficient for the complete protection of the commercially available chickens vaccinated at the age of 5 wk and challenged 7 wk later. The anti-NDV-free chickens, vaccinated at 4 wk of age and challenged 2 wk postvaccination, were 100% and 73% protected by a vaccinal dose containing 178.6 and 89.3 microliters of detergent-treated NDV-AF, respectively. Commercially available light pullets, primary vaccinated with live lentogenic NDV vaccine, generated a protective level of NDV antibodies after revaccination with WOWE vaccine containing 89.3 microliters of detergent treated NDV-AF. Laying hens were revaccinated under field conditions at the beginning of the laying cycle by the tetravalent vaccine. A vaccinal dose/bird containing 11.2 microliters of detergent-treated NDV-AF elicited a long-lasting high level of NDV neutralizing antibodies. PMID- 8713035 TI - Multicentric histiocytosis in young chickens. Gross and light microscopic pathology. AB - During 1991-94, tissue specimens from 262 young chicken carcasses condemned at slaughter contained novel multicentric proliferations of histiocytelike cells. These tissues had been submitted to the USDA FSIS Eastern Laboratory because of grossly enlarged spleens, livers, or kidneys. The spleens were two to three times normal diameter and contained miliary white or yellow 1-3-mm foci. Similar miliary foci were present throughout the enlarged livers and kidneys. Microscopic examination of these tissues revealed discrete circular nodules expanding splenic periarteriolar lymphoid sheaths, hepatic periportal nodules, and discrete perivascular and more diffuse interstitial nodules replacing renal tubules. Nodules also were present in the pancreas, bone marrow, proventriculus, and lung, with more diffuse infiltrates in intestinal lamina propria. The cells composing these nodules contained irregularly oval, folded, or pleomorphic nuclei and relatively abundant eosinophilic cytoplasm. Mitotic figures and pyknotic nuclei were common. These cells were interpreted to be histiocytes (tissue macrophages or dendritic cells) and did not resemble lymphocytes. These proliferating cells also did not resemble the cell population of commonly diagnosed lymphoid neoplasms of young chickens. No intralesional organisms were detected and polymerase chain reaction analysis failed to detect Marek's herpesvirus DNA or leukosis/sarcoma and reticuloendotheliosis proviral DNA. PMID- 8713036 TI - Influence of chicken breed on pathogenicity evaluation of velogenic neurotropic Newcastle disease virus isolates from cormorants and turkeys. AB - Three isolates of Newcastle disease virus (NDV) from cormorants and turkeys were classified as velogenic neurotropic NDV (VNNDV) by standard pathotyping procedures in specific-pathogen-free white leghorn and white rock chickens. White leghorns inoculated by eye drop, cloaca, intravenous, or intracerebral routes had a higher frequency of nervous signs and mortality than was observed in similarly inoculated white rocks. Two backpassages in white rocks increased the intravenous pathogenicity index of all three isolates, but none had a value after backpassage that was as high as the classical VNNDV strain Texas GB. The breed of chicken used in standard procedures did influence the numerical values of pathotyping determinations, but the differences between breeds were not large enough to change the pathotype assigned to these isolates. PMID- 8713037 TI - Polymerase chain reaction optimization for Mycoplasma gallisepticum and M. synoviae diagnosis. AB - Mycoplasma gallisepticum- or M. synoviae-challenged chickens were monitored with serological assays (serum plate agglutination, hemagglutination inhibition, and enzyme-linked immunosorbent assay) and polymerase chain reaction (PCR). The tracheal swabs from M. gallisepticum-challenged chickens received three different treatments (phosphate-buffered saline [PBS], Frey's broth, or 10 mM Tris-HCl/250 mM ethylenediaminetetraacetic acid/ 2.5% sodium dodecyl sulfate [STE]) prior to DNA purification. A nonphenolic method for DNA extraction was utilized. The best PCR results were obtained with PBS swab treatment. The nonphenolic method for DNA extraction was compared with a phenolic method in an experiment with tracheal swabs from M. synoviae-challenged chickens and commercial flocks. Both methods gave comparable results. PMID- 8713038 TI - Acute cardiotoxicity of moniliformin in broiler chickens as measured by electrocardiography. AB - Electrocardiography was used to examine the acute cardiotoxic effects of moniliformin on 3-week-old broiler chickens. Each of the seven pairs of anesthetized birds (pentobarbital sodium, 40 mg/kg body weight, intramuscular) was injected intravenously with moniliformin (1 mg/kg body weight) or an equal volume of normal saline (1 ml/kg body weight), and changes in electrocardiogram were monitored for 50 minutes. Three of the seven birds injected with moniliformin died within 50 minutes post-injection. Moniliformin caused a bradycardia, which became highly significant (P < 0.05) within 15 minutes post injection. The P-R, Q-T, and S-T intervals of moniliformin-injected birds were significantly lengthened throughout the 50-minute observation (P < 0.05). The results indicate that the moniliformin-induced mortality is due primarily to cardiac failure. PMID- 8713039 TI - Lysis of erythrocytes by Trichomonas gallinae. AB - The hemolytic activity of five live isolates of Trichomonas gallinae was investigated. The isolates were subsequently tested against the erythrocytes of seven adult animal species. Each of the five isolates tested lysed all human blood groups, as well as rabbit, rat, chicken, horse, bovine, and sheep erythrocytes. No hemolysin released by the parasite could be detected. Our preliminary results suggest that the hemolytic activity is not due to the hemolysin release by T. gallinae or to a product of its metabolism. Pretreatment of live trichomonads with concanavalin A reduced levels of hemolysis by 40%. PMID- 8713040 TI - Receptivity and susceptibility of the domestic duck (Anas platyrhynchos), the Muscovy duck (Cairina moschata), and their hybrid, the mule duck, to an experimental infection by Eimeria mulardi. AB - The receptivity and susceptibility of the Muscovy duck, the domestic duck, and their hybrid, the mule duck, toward an experimental infection by Eimeria mulardi were studied. To appraise their susceptibility, female ducklings were infected with a dose of 75,000 oocysts per duckling (bringing about a clinical coccidiosis). In another group, to compare the multiplication rates of the parasite in the three species of ducks, thus assessing their receptivity, an infection with 750 oocysts per duckling was realized. Eimeria mulardi develops in the three species of ducks. In each case, a significant decrease in growth in the contaminated group vs. the control group was noted, even though the Muscovy and the mule ducks were more susceptible to the contamination. PMID- 8713041 TI - The chronic effects of Fusarium moniliforme culture material, containing known levels of fumonisin B1, in turkeys. AB - Fourteen 1-day-old male turkeys were randomly assigned to two adjacent floor pens and fed balanced rations containing 0 and 75 mg fumonisin B1 (FB1)/kg for 18 weeks. Inclusion of FB1 in the ration caused decreased body weight gain on weeks 4, 10, and 12 during the trial. Turkeys fed 75 mg FB1/kg had significantly heavier livers after treatment for 18 weeks. Chronic FB1 exposure resulted in an increased total white blood cell count, absolute heterophil count, absolute lymphocyte count, and heterophil-to-lymphocyte ratio. No mortality was noted in turkeys in either treatment group. Turkeys are relatively resistant to chronic FB1 exposure. PMID- 8713043 TI - Causes of losses including a Borna disease paralytic syndrome affecting young ostriches of one breeding organization over a five-year period (1989-1993). AB - Necropsy records and causes of mortality of ostriches up to 3 months old over a 5 year period (1989-1993) are presented. The data relate to one ostrich enterprise that comprises 10 breeding flocks, five rearing farms, and one hatchery. Causes of mortality are classified into nine major categories. The annual mortality percentages of all hatched ostriches over the 5-year period were 61%, 58%, 30%, 29%, and 16.6%, and the most significant cause of death was a paresis syndrome that accounted for 20%, 11%, 16%, 10.1%, and 2% mortality, respectively. Limb deformities and gastroenteritis were the other principal specific causes of mortality. The paresis syndrome was caused by an agent serologically related to Borna disease virus. Brain extracts from paralyzed ostriches, when given orally or intramuscularly to 5-week-old birds, reproduced the clinical signs and microscopic lesions. The mean time to death was less than 3 weeks for the intramuscularly infected group and was almost twice as long for the orally infected group. PMID- 8713042 TI - Copper sulfate toxicosis in commercial laying hens. AB - A flock of 51-week-old leghorn hens experienced a 16% drop in egg production in a single week. The layer ration contained 1477 ppm copper from the addition of copper sulfate. Severe oral ulcers were present in the pharynx. Oral ulcers, reduced feed intake, and a drop in egg production occurred when a ration containing 1437 ppm copper was evaluated experimentally. PMID- 8713044 TI - Mycobacterium genavense infection in canaries. AB - A case of mycobacteriosis in a collection of canaries (Serinus canarius) is described. The affected birds showed nonspecific clinical signs and macroscopic lesions (slight splenomegaly). Histologic lesions found in liver, spleen, lungs, and kidneys consisted of noncaseous nodules containing accumulations of large macrophages that showed a highly vacuolated cytoplasm in which numerous acid-fast organisms were detected. Attempts to isolate and culture the organisms using Lowenstein-Jensen and Coletsos media proved unsuccessful. Microorganisms belonging to the species Mycobacterium genavense were identified by means of polymerase chain reaction techniques in hepatic tissue from both birds. This finding confirms the existence of fastidiously growing mycobacterial infections, other than Mycobacterium avium, in birds. PMID- 8713045 TI - The effect of bacterial surface structures on the pathogenesis of Salmonella typhimurium infection in chickens. AB - Mutants of Salmonella typhimurium strain SR11 containing Tn10 insertions in genes encoding for the motility and fimbriation phenotypes were evaluated for adhesion, invasion, and virulence in avian models. Mutations abolishing mannose-sensitive or mannose-resistant hemagglutination did not influence adherence or invasion in epithelial cells in vitro. A double hemagglutinin-deficient mutant, lacking both mannose-sensitive and mannose-resistant hemagglutinins, was diminished in ability to adhere to chick kidney epithelial cells in vitro, but invasion in vitro was not significantly affected. Compared with the wild-type parent, mutations that decreased motility reduced invasion levels in vitro and increased the peroral LD50 in one-day-old chicks. A mutant deficient in both motility and mannose sensitive hemagglutination was greatly reduced in its ability to invade epithelial cells in vitro and persist in the liver and spleen of orally challenged chicks. Results of this study indicate that loss of motility in S. typhimurium attenuates peroral virulence in chicks. PMID- 8713046 TI - Immunity to eimeria tenella in chickens: phenotypical and functional changes in peripheral blood T-cell subsets. AB - The changes in peripheral blood leukocyte (PBL) T-cell subsets following Eimeria tenella infection in outbred white leghorn chickens were studied, using a panel of murine monoclonal antibodies specific for the chicken homologues of the mammalian CD3, CD8, and CD4 markers on day-to-day samples of PBLs. Both flow cytometric analysis (FCA) and immunofluorescence microscopy with fixed cells on slides were used as read-out systems. The changes in the composition of the T cell subsets measured with both techniques were similar. At 8 days post primary infection, a sharp transitory increase in the proportion of CD8-expressing cells was found. With FCA, CD8-expressing cells could be discriminated in CD8(Dim+) and CD8(Bright+) populations, which have not been described before. The proportion of CD4-expressing cells was decreased at days 9-10 after primary infection, which coincided with a less marked decrease in CD3-expressing cells. Such effects were not seen after secondary infection. When PBLs collected at day 8 post primary infection were stimulated in vitro with E. tenella sporozoite antigen, the response was higher than that in uninfected control chickens. The effects we observed coincide with the onset of recovery from primary infection. We speculate that the increase in CD8-expressing PBLs is the result of stimulation and expansion of a specific subset involved in the induction of protective immunity against Eimeria tenella. PMID- 8713047 TI - Characterization of monoclonal antibodies against infections laryngotracheitis virus. AB - Nine monoclonal antibodies (MCAs) produced against two different strains of infectious laryngotracheitis virus (ILTV) were characterized and compared to previously characterized MCA 131-6, produced against a third ILTV strain. In western blotting experiments, MCAs C, E, and 11 resembled MCA 131-6, detecting proteins of 205, 160, 115, and 90 kD as well as several proteins less than 49 kD. The other six MCAs differed from previously described ILTV MCA. MCA D detected a 90-kD protein along with several less than 49 kD. MCAs 4 and 5 each detected proteins of 205, 160, 100, 90, and 70 kD. MCA 9 detected the same proteins detected by MCAs 4 and 5 except the 160-kD protein. MCA 10 detected proteins of 100, 90, and 70 kD and several proteins less than 49 kD. MCAs C, D, and E, like MCA 131-6, failed to react with any ILTV grown in the presence of tunicamycin, suggesting that those MCAs are specific for carbohydrate-based epitopes. MCA 6 reacted with only a 100-kD protein in the presence or absence of tunicamycin. The remaining MCA detected only a 70-kD protein in the presence of tunicamycin except MCA 5, which reacted with proteins of 70 and 90 kD. Only MCA 4 and 6 neutralized ILTV infectivity. PMID- 8713048 TI - Development of a polymerase chain reaction and a nonradioactive DNA probe for infectious laryngotracheitis virus. AB - The polymerase chain reaction (PCR) was developed using infectious laryngotracheitis virus (ILTV) primers made from a portion of the ILTV thymidine kinase gene. DNA from various ILTV field isolates, from the USDA challenge strain of ILTV, and from commercial ILTV vaccines was specifically amplified. No amplification occurred using template DNA from uninfected chicken-embryo liver cells (CELC), several nonavian alphaher-pesviruses, Mycoplasma gallisepticum, Mycoplasma synoviae, Pasteurella hemolytica, Escherichia coli, a group I avian adenovirus, fowl poxvirus, or a psittacid herpesvirus. The 647-base pair amplified ILTV PCR product was labeled to create a nonradioactive, biotinylated DNA probe. Hybridization using the probe detected ILTV DNA. Both PCR and hybridization yielded positive results with ILTV DNA but not with the DNA of other pathogens. Hybridization was specific for ILTV using a stringent salt solution for a 30-min wash step or a somewhat less stringent salt solution for a 60-min wash step. However, slight hybridization occurred with CELC DNA when the less stringent salt solution was used in a 30-min wash step. PMID- 8713049 TI - Increasing the yield of Eimeria tenella oocysts in primary chicken kidney cells. AB - The present study was undertaken to increase the yield of Eimeria tenella oocysts in primary chicken kidney cells (PCKCs) using a comparatively inexpensive cell culture system. PCKCs growing on coverslips positioned on the bottoms of individual wells in 24-well tissue-culture plates were infected with sporozoites of E. tenella. The effects of changing the culture medium (RPMI 1640), medium pH, serum type, and serum concentration in the wells were determined by counting newly produced oocysts at 7 days postinoculation. There were significantly more (P < 0.01) oocysts produced when the medium was supplemented with 10% fetal bovine serum (FBS) and changed either daily or every other day compared with not changing the medium. When the same medium was changed daily, significantly more (P < 0.05) oocysts were produced at pH 7.4 than at pH 8.0 but not at pH 6.0. If the medium was changed daily, significantly more (P < 0.05) oocysts were produced when medium was supplemented with 10% FBS only rather than 5% and 10% chicken serum. The cell-culture system described in this study offers a convenient and efficient method for investigating the biological, biochemical, and immunological relationships between E. tenella and the host cell. PMID- 8713050 TI - Effect of semduramicin, salinomycin, and monensin on performance, shank pigmentation, and coccidial lesions in broiler chickens in floor pens. AB - The effects of 25 ppm semduramicin, 66 ppm salinomycin, 110 ppm monensin, and unmedicated treatments on performance, shank pigmentation, and coccidial lesion scores in broiler chickens were evaluated in two floorpen trials in the United States. On day 24 of each test, birds in each treatment were inoculated via the feed with a mixture of recent field isolates of Eimeria spp. at a dose rate calculated to provide 2 x 10(5) E. acervulina, 3 x 10(4) E. maxima, and 2 x 10(4) E. tenella sporulated oocysts per bird. Weight gain and feed conversion were significantly (P < or = 0.05) improved in the semduramicin-treated broilers in comparison with the monensin-treated and unmedicated broilers. These performance variables for the salinomycin-treated birds were intermediate between the semduramicin- and monensin-treated birds. Shank pigmentation scores were significantly (P < or = 0.05) improved in the three anticoccidial treatments compared with unmedicated birds, with the highest scores (P < or = 0.05) occurring in the semduramicin-treated broilers. Semduramicin was more efficacious (P < or = 0.05) than salinomycin in controlling upper intestinal lesions and more efficacious than monensin in controlling mid-intestinal lesions. All three drugs were comparable in controlling lesions in the ceca. PMID- 8713051 TI - Heat destruction of Salmonella in poultry feed: effect of time, temperature, and moisture. AB - Salmonella enteriditis in poultry feed declines with increasing time of exposure to heat. The interactions of temperature, time, and moisture and their effect on the thermal death of S. enteriditis were established in a factorial randomized experiment. Two other serotypes were tested, and though there was some variation, the thermal death rate followed the same basic pattern. A number of samples of poultry feed were collected and dried. After drying, the water was added back to give specific percentages of moisture contents. The feed was then inoculated with salmonella and heated at specific temperatures, with samples being removed at certain time intervals. These samples were then cultured, and the surviving salmonella were counted. A linear relationship was obtained when the logarithm of survivors was plotted against the logarithm of exposure time. These results permitted the construction of a graph depicting that the rate of reduction in numbers of S. enteriditis when plotted against increasing temperatures is linear. This linear relationship is apparent for other salmonella serotypes such as S. typhimurium and S. haardt. Our results show that the thermal death rate of salmonella in poultry feed can now be predicted at varying time, temperature, and moisture contents. PMID- 8713052 TI - Interferon modulation of Marek's disease virus genome expression in chicken cell lines. AB - Lines of chicken lymphoblastoid cells were established from local lesions induced by simultaneous injection of Marek's disease virus and various stimulants of T cell activation. Lines developed with regular medium had relatively high mean rates of spontaneous expression of viral internal antigen (6.2%). In contrast, lines developed and maintained with conditioned medium generated by mixed lymphocyte reaction had a 62-fold reduction in the mean rate of viral internal antigen expression (0.1%). The expression rate could be modulated by the removal or re-addition of conditioned medium to the growth medium. Down regulation involved proteins classified as immediate-early (a 14-kDa polypeptide), early (a 38-kDa phosphoprotein), and late (glycoprotein B homologue) antigens, indicating that the block is very early in virus replication. Once initiated in a given cell, replication apparently proceeded unimpeded. Interferon was determined to be largely responsible for the suppressive activity of the conditioned medium, although involvement of other cytokines could not be ruled out. Also, chicken interferon from other sources, including recombinant interferon, was able to similarly suppress viral antigen expression. PMID- 8713053 TI - Immunohistochemical study of a chondroitin-6-sulfate in growth plates of broiler chickens with high and low genetic predispositions to tibial dyschondroplasia. AB - The distribution of a chondroitin-6-sulfate (C6S) epitope, which is a biochemical marker of chondrocyte hypertrophy, was studied in the growth plates of two lines of 3-week-old broiler chickens with low and high genetic predispositions to tibial dyschondroplasia (TD). Ultrathin sections of growth plates from both groups were subjected to immunolocalization with monoclonal antibody 3-B-3(-), the epitope of which is increased on proteoglycans made by hypertrophic chondrocytes. Bound antibody was localized with colloidal gold-labeled protein A for observation with an electron microscope. The 3-B-3(-) epitope was localized in pericellular and interterritorial matrix of growth plates of both lines. In the low-TD-incidence birds, the concentration of 3-B-3(-) bound to C6S progressively increased from the proliferative zone to the hypertrophic zone. However, in the high-TD-incidence line, the epitope expression remained at a low level in all zones. The increase of the 3-B-3(-) epitope produced by maturing growth-plate chondrocytes is indicative of changes in the glycosaminoglycan chains of proteoglycans that may be important in the process of matrical calcification. Thus, failure of chondrocytes of the high-TD-incidence line to produce this change in post-translational modification of their proteoglycans could be important in the pathological process. PMID- 8713054 TI - A pathogenic challenge model for adult northern bobwhite (Colinus virginianus) using a vaccine strain of Pasteurella multocida type 3. AB - A pathogenic challenge model causing approximately 50% mortality was developed in adult Northern bobwhite (Colinus virginianus) using Avichol, a live vaccine containing the Clemson University (CU) strain of Pasteurella multocida Type 3. A dose of 2300 or 3000 colony-forming units (CFU) of Avichol injected intramuscularly resulted in 30 to 75% mortality, whereas a dose of 230 CFU or less resulted in no mortality, and 58,720 CFU or more resulted in death in all birds challenged. Primary and secondary vaccination of Northern bobwhite with a formalinized anaculture of Avichol -derived P. multocida resulted in protection against challenge in three separate experiments. Dexamethasone treatment of birds during vaccination resulted in decreased protection against challenge exposure. PMID- 8713056 TI - Phosphatidylserine-mediated adhesion of T-cells to endothelial cells. AB - Phosphatidylserine was exposed on the surface of human umbilical endothelial cells (ECV304) a few minutes after adding thrombin in vitro, as monitored by prothrombinase assays with and without annexin V. Jurkat T cells adhered to the thrombin-treated cells. The adhesion was inhibited by annexin V, indicating that it was mediated by exposed phosphatidylserine on the endothelial cells. PMID- 8713057 TI - Thrombin receptors modulate insulin-stimulated phosphatidylinositol 3,4,5 trisphosphate accumulation in 1321N1 astrocytoma cells. AB - Thrombin and insulin receptor signaling via phosphoinositide (PI)-specific phospholipase C (PLC) and PI 3-kinase was studied in [3H]inositol-labelled 1321N1 cells. Thrombin stimulated a dramatic, transient activation of PLC which is probably mediated via receptors of the 'tethered-ligand' type, since it was both reproduced by, and abolished following, pretreatment of cells with a synthetic peptide (SFLLRN) corresponding to the ligand domain of the human thrombin receptor. However, neither thrombin nor SFLLRN stimulated PI 3-kinase. By contrast, insulin did not influence [3H]InsP3 concentration but stimulated accumulation of [3H]PtdIns(3,4,5)P3 and [3H]PtdIns(3,4)P2, the relative steady state concentrations of which may indicate degradation of [3H]PtdIns(3,4,5)P3 by 5- and 3-phosphatases. The independent coupling of thrombin and insulin receptors to PLC and PI 3-kinase respectively in 1321N1 cells allowed interactions between these systems to be examined. Thus insulin-stimulated [3H]PtdIns(3,4,5)P3 accumulation was attenuated on co-stimulation of the thrombin receptor, whereas concentrations of [3H]PtdIns(3,4)P2 were transiently enhanced but then reduced. These results indicate that thrombin receptors in 1321N1 cells do not activate PI 3-kinase, but can modulate signalling by this enzyme. PMID- 8713058 TI - Hexose uptake in Trypanosoma cruzi: structure-activity relationship between substrate and transporter. AB - The gene encoding a hexose transporter, TcrHt1, from Trypanosoma cruzi has been functionally expressed in mammalian Chinese hamster ovary cells. Kinetic parameters of the heterologously expressed protein are very similar to those of the transporter identified in T. cruzi epimastigotes, confirming that TcrHT1 is the major transporter functioning in these parasites. A detailed analysis of substrate recognition using analogues of D-glucose substituted at each carbon position has been performed. The glucose transporter of T. cruzi does not recognize C-3 or C-6 analogues of D-glucose, whereas these analogues were recognized by the glucose transporter of bloodstream-form T. brucei. As for other kinetoplastid transporters, but in stark contrast to the mammalian GLUT family, TcrHT1 can also transport D-fructose, with relatively high affinity (Km = 0.682 +/- 0.003 mM). Amino acid side-chain-modifying reagents were also used to identify residues of the transporter present at the substrate-binding site. While specific modifiers of cysteine, histidine and arginine all inhibited catalytic activity, protection using substrate was only observed using the arginine specific reagent, phenylglyoxal. Reagents which modify lysine residues had no effect on transport. PMID- 8713059 TI - Effect of dietary protein restriction on liver transcription factors. AB - The transcription of several genes that are preferentially expressed in the liver, including the serum albumin, transthyretin and carbamyl phosphate synthetase-I genes, is specifically decreased in animals consuming inadequate amounts of dietary protein. The high level of transcription of these genes in the liver is directed in part by a number of liver-enriched transcription factors, including hepatocyte nuclear factors (HNF)-1, -3, and -4, and proteins of the CCAAT/enhancer-binding protein (C/EBP) family. In the present study, we investigated the possibility that the co-ordinate decrease in transcription of the nutritionally sensitive genes in protein-deprived rats results from altered activity of one or more of the liver-enriched transcription factors. For HNF-4, Western blots indicated no change in the level of nuclear HNF-4 protein in liver of protein-deprived animals, whereas we observed a 40% reduction in the DNA binding activity of HNF-4 as measured by electrophoretic mobility shift assay (EMSA). Furthermore, the binding affinity of HNF-4 for DNA was unaltered by dietary protein deprivation, while the number of HNF-4 molecules able to bind to DNA (Bmax) was reduced, as determined by Scatchard analysis. This indicates that in the protein-restricted rats a portion of the pool of HNF-4 protein is inactivated or otherwise prevented from binding to DNA. The overall DNA binding activity of C/EBP alpha and beta was increased in protein-restricted animals. This change occurred in the absence of a change in the amount of the full-length forms of these two proteins, quantified by Western blotting. Interestingly, dietary protein restriction specifically increased the level of a truncated form of C/EBP beta (liver-enriched transcriptional inhibitory protein, LIP), which is a protein dominant negative inhibitor of C/EBP function. Analysis of HNF-3 DNA binding activity by EMSA revealed that HNF-3 alpha and beta DNA binding was increased and that HNF-3 gamma DNA-binding activity was unchanged in protein restricted animals. We also detected two apparently novel shift complexes with the HNF-3 probe by EMSA, both of which were decreased in protein-restricted animals. HNF-1 DNA-binding activity was increased by dietary protein restriction. We also examined the effect of protein restriction on the DNA-binding activity of two ubiquitous transcription factors, NF1 and Sp1. The DNA binding activity of the major NF1 isoforms was unchanged whereas the binding activity of Sp1 was increased in the protein-restricted animals. In summary, restriction of dietary protein resulted in a number of specific changes in the DNA-binding activity of various transcription factors. Because transcriptional activation typically involves the synergistic action of more than one transcription factor, small changes in the amount/activity of several factors, could have a strong net effect on the transcription of many genes. PMID- 8713055 TI - Multiple steps in the regulation of transcription-factor level and activity. AB - This review focuses on the regulation of transcription factors, many of which are DNA-binding proteins that recognize cis-regulatory elements of target genes and are the most direct regulators of gene transcription. Transcription factors serve as integration centres of the different signal-transduction pathways affecting a given gene. It is obvious that the regulation of these regulators themselves is of crucial importance for differential gene expression during development and in terminally differentiated cells. Transcription factors can be regulated at two, principally different, levels, namely concentration and activity, each of which can be modulated in a variety of ways. The concentrations of transcription factors, as of intracellular proteins in general, may be regulated at any of the steps leading from DNA to protein, including transcription, RNA processing, mRNA degradation and translation. The activity of a transcription factor is often regulated by (de) phosphorylation, which may affect different functions, e.g. nuclear localization DNA binding and trans-activation. Ligand binding is another mode of transcription-factor activation. It is typical for the large super-family of nuclear hormone receptors. Heterodimerization between transcription factors adds another dimension to the regulatory diversity and signal integration. Finally, non-DNA-binding (accessory) factors may mediate a diverse range of functions, e.g. serving as a bridge between the transcription factor and the basal transcription machinery, stabilizing the DNA-binding complex or changing the specificity of the target sequence recognition. The present review presents an overview of different modes of transcription-factor regulation, each illustrated by typical examples. PMID- 8713060 TI - The respiratory-chain poison antimycin A promotes the formation of DNA single strand breaks and reduces toxicity in U937 cells exposed to t-butylhydroperoxide. AB - Antimycin A at levels that abolish oxygen consumption had a slight, although statistically significant, inhibitory effect on the toxicity elicited by t butylhydroperoxide in U937 cells. The protective effect was observed after 6 h of post-treatment incubation, but was no longer apparent after 24 h. Unexpectedly, these events were associated with a marked accumulation of DNA single-strand breaks produced by low levels of t-butylhydroperoxide. Both an oxygen- and a carbon-centred radical were found to arise during treatment with t butylhydroperoxide, and their formation was significantly lowered by antimycin A. Thus inhibition of electron transport at the level of complex III appears (a) to decrease the formation of toxic species which mediate, at least partially, the lethal effects elicited by t-butylhydroperoxide, and (b) to enhance the formation of DNA-damaging species generated at low concentrations of t-butylhydroperoxide. Rotenone and cyanide, which respectively inhibit complexes I and IV, did not affect DNA damage elicited by t-butylhydroperoxide. These results suggest that DNA single-strand breaks do not mediate the toxicity of t-butylhydroperoxide, and that specific mitochondrial functions might modulate the formation of the toxic and of DNA-damaging species generated by organic hydroperoxides. PMID- 8713061 TI - Activation by phosphorylation of phosphofructokinase from the annelid Lumbricus terrestris and comparison of phosphorylated sites in invertebrate phosphofructokinases. AB - Purified phosphofructokinase from the earthworm Lumbricus terrestris was phosphorylated in vitro by the catalytic subunit of cAMP-dependent protein kinase from the same organism to an extent of approx. 0.5 mol/mol of subunit. Activation of the enzyme occurred in parallel to the incorporation of covalently bound phosphate and was reversed by the action of the catalytic subunit of protein phosphatase 2A. Phosphorylation decreased the co-operativity of fructose 6 phosphate saturation in the presence of inhibitory concentrations of ATP, and increased the apparent Vmax obtained with saturating concentrations of the activators 5'-AMP and fructose 2,6-bisphosphate. The phosphorylated sites of phosphofructokinase from L. terrestris and from two molluscs (Helix pomatia and Mytilus edulis) were sequenced and shown to exhibit distinct similarity to sequences located near to the N-terminus of nematode phosphofructokinases [Klein, Olson, Favreau, Wintertowed, Hatzenbuhler, Shea, Nulf and Geary (1991) Mol. Biochem. Parasitol. 48, 17-26. PMID- 8713062 TI - Apoptosis induction resulting from proteasome inhibition. AB - Proteases are known to be involved in the apoptotic pathway. We report here that benzyloxycarbonyl (Z)-Leu-Leu-leucinal(ZLLLal), a leupeptin analogue, can induce apoptosis in MOLT-4 and L5178Y cells. ZLLLal is a cell-permeant inhibitor of proteasome. Among the protease inhibitors tested, only calpain inhibitor I (acetyl-Leu-Leu-norleucinal) and ZLLLal caused a marked induction of apoptosis in MOLT-4 cells. In contrast Z-Leu-leucinal, a specific inhibitor of calpain, did not induce apoptosis. When MOLT-4 cells were incubated in the presence of ZLLLal, p53 accumulated in the cells. These results strongly suggest that inhibition of proteasome induces p53-dependent apoptosis and that proteasome can protect cell from apoptosis. PMID- 8713063 TI - Characterization of calf liver Cu,Zn-metallothionein: naturally variable Cu and Zn stoichiometries. AB - Cu,Zn-metallothioneins were purified from bovine calf liver in order to examine the stoichiometry of metal binding to the protein. Copper and zinc analyses were carried out by atomic absorption spectrophotometry. Consistent quantitative thiolate analyses were obtained spectrophotometrically with Ellman's reagent and amperometrically with phenylmercuric acetate. These were used to define protein concentration. A complementary method to assess the sum of the thiol and Cu(I) content of metallothionein involved titration of the reducing equivalents of the protein with ferricyanide. The stoichiometry of reaction was consistent with the oxidation of all the sulphydryl groups to disulphides and all of the bound Cu from the cuprous to the cupric oxidation state. Accordingly to these methods, total numbers of zinc plus copper ions bound to metallothionein isolated from a number of calf livers centred on about 7, 10-12, or 15 g-atoms of metal per mol of protein. The reactivity of ferricyanide and 4,7-phenylsulphonyl-2,9-dimethyl 1, 10-phenanthroline (BCS) with Cu,Zn-metallothioneins of various metal ratios was assessed. Zinc metallothionein reacted almost entirely in two slow steps with ferricyanide. As the Cu content of the protein increased, the fraction of reaction occurring in the time of mixing increased in parallel. BCS was able to remove 70-80% of metallothionein-bound Cu as Cu(I). The rest was resistant to reaction. PMID- 8713064 TI - Stoichiometry and cluster specificity of copper binding to metallothionein: homogeneous metal clusters. AB - Experiments were done to define the stoichiometry of binding of Cu(I) to metallothionein (MT) and to determine its sites of binding in mixed-metal species. Spectrophotometric titrations of rabbit liver Cd7-MT 2, apoMT, and Cd4 alpha-domain with Cu(I) revealed endpoints of 3-4, 4 and 8, and 4 and 6-7 added Cu(I)/mol of MT for the three species respectively. Observed endpoints depended on conditions of the titration and the wavelength chosen for absorbance measurement. Nevertheless, from metal and sulphydryl analyses of titrated proteins that were pretreated with Chelex-100 to remove metal ions from solution, almost all of the cadmium was displaced from Cd7-MT by the addition of 7 Cu(I)/mol of MT. Similarly, 4 Cu(I)/mol of Cd4-alpha-domain completely displaced bound cadmium. The Cu4-alpha-domain was converted into a Cu6-alpha species upon addition of two equivalents of Cu(I)/mol of alpha-domain. Reaction of Cd7-MT with 7, 12 and 20 Cu/mol of MT, followed by reaction with Chelex resin, generated protein samples in each case with about 7 Cu/mol of MT. 111Cd-NMR analysis of the reaction of 111Cd7-MT with Cu(I) showed that nearly co-operative one-for-one replacement of 111Cd occurred and that the beta-domain cluster reacted before the alpha-domain cluster. Two mixed-metal MTs with Cu to Zn ratios approximating 3 to 4 and 6 to 4 were isolated from calf liver. After substitution of Zn with 111Cd, NMR spectra of each protein showed that 111Cd was confined almost completely to the alpha-domain. By inference, about 3 or 6 Cu were bound in the beta-domain of these proteins. Supporting this segregation of metal ions into domains, reaction of Cu6, Zn4-MT with nitrilotriacetate removed zinc exclusively, whereas reaction of Cu6,Cd4-MT with 4,7-phenylsulphonyl-2,9-dimethyl-1,10-phenanthroline extracted only Cu(I). Proteolytic digestion of both products followed by gel filtration demonstrated that Cu(I) and Cd were bound to fragments of the intact protein. Finally, reaction of rabbit liver 111Cd7-MT 2 with Cu10-MT 2 resulted in interprotein metal exchange in which 111Cd-moved from the beta- to the alpha domain according to NMR analysis. In contrast with the prevalent view that six Cu(I) bind to each domain of MT, the present results show that Cu(I) binds to MT with a minimum stoichiometry of about 7 Cu(I)/mol of MT and can bind to the alpha domain with stoichiometries of 4 or 6 Cu(I)/mol of MT. Although MTs interacting with 12 or 20 Cu(I)/mol of MT are less stable than that binding about 7 Cu(I)/mol, it appears that MT can bind Cu(I) in multiple stoichiometries. PMID- 8713065 TI - Ca2+ signalling mechanisms of the beta 2 integrin on neutrophils: involvement of phospholipase C gamma 2 and Ins(1,4,5)P3. AB - Engagement of beta 2 integrins triggers a tyrosine kinase-dependent intracellular mobilization and influx of Ca2+ in human neutrophils. However, the transduction pathway involved in generating this Ca2+ signal is obscure. In the present study we identified phospholipase C gamma 2 (PLC gamma 2) as one of the major proteins that was phosphorylated on tyrosine in response to beta 2 integrin activation. This beta 2 integrin-induced phosphorylation of PLC gamma 2 occurred in parallel with an increased accumulation of Ins(1,4,5)P3. The relevance of these observations for the beta 2 integrin-induced Ca2+ signal was investigated using an inhibitor of PLC signalling pathways, 1-(6-{[17 beta-3-methoxyoestra-1,3.5(10) trien-17-yl] amino}hexyl)-1H-pyrrole-2,5-dione(U73122). U73122 dose-dependently (IC50, approx. 0.15 microM) inhibited both the beta 2 integrin-induced release of Ca2+ from intracellular stores and the subsequent influx of Ca2+ across the plasma membrane. These effects were not observed with the inactive analogue 1-(6 {[17 beta-3-methoxyoestra-1,3,5(10)-trien-17-yl] amino}hexyl)-pyrrolidine-2,5 dione (U73343). To gain further support for an involvement of PLC-induced Ins(1,4,5)P3 formation in the beta 2 integrin-induced Ca2+ signal, we searched for the molecular event(s) underlying the effects of U73122. Our experiments revealed that U73122 had no effect on either beta 2 integrin-induced tyrosine phosphorylation of PLC gamma 2 (or any of the other proteins) or on the formation of Ins(1,4,5)P3, but it reduced the Ins(1,4,5)P3-induced release of 45Ca2+ from intracellular stores of electropermeabilized cells. Taken together, the present data suggest that the beta 2 integrin-induced Ca2+ signal in human neutrophils is generated through activation of a PLC gamma 2-dependent pathway. PMID- 8713066 TI - Alteration in gene expression of branched-chain keto acid dehydrogenase kinase but not in gene expression of its substrate in the liver of clofibrate-treated rats. AB - We previously showed that the oxidation of branched-chain amino acids is increased in rats treated with clofibrate [Paul and Adibi (1980) J. Clin. Invest. 65, 1285-1293]. Two subsequent studies have reported contradictory results regarding the effect of clofibrate treatment on gene expression of branched-chain keto acid dehydrogenase (BCKDH) in rat liver. Furthermore, there has been no previous study of the effect of clofibrate treatment on gene expression of BCKDH kinase, which regulates the activity of BCKDH by phosphorylation. The purpose of the present study was to investigate the above issues. Clofibrate treatment for 2 weeks resulted in (a) a 3-fold increase in the flux through BCKDH in mitochondria isolated from rat liver, and (b) a modest but significant increase in the activity of BCKDH. However, clofibrate treatment had no significant effect on the mass of E1 alpha, E1 beta, and E2 subunits of BCKDH or the abundance of mRNAs encoding these subunits. On the other hand, clofibrate treatment significantly reduced the activity, the protein mass and the mRNA levels of BCKDH kinase in the liver. In contrast to the results obtained in liver, clofibrate treatment had no significant effect on any of these parameters of BCKDH kinase in the skeletal muscle. In conclusion, our results show that clofibrate treatment increases the activity of BCKDH in the liver and the mechanism of this effect is the inhibition of gene expression of the BCKDH kinase. PMID- 8713067 TI - Differential regulation of neuronal nicotinic acetylcholine receptor subunit gene promoters by Brn-3 POU family transcription factors. AB - The regulatory region of the neuronal nicotinic acetylcholine (nACh) receptor alpha 2 subunit gene is activated by the Brn-3b POU family transcription factor but not by the closely related factors Brn-3a and Brn-3c. This pattern of regulation has not previously been observed for other neuronally expressed genes, several of which, such as those encoding alpha-internexin or SNAP-25, are activated by Brn-3a and Brn-3c but repressed by Brn-3b. The alpha 3 nACh receptor subunit gene is also shown to be activated by Brn-3a but is repressed by Brn-3b and Brn-3c. In contrast, the Brn-3 POU family transcription factors have no effects on either the alpha 7 or beta 4 nACh receptor subunit genes. The actions of Brn-3b on the alpha 2 subunit are thus in contrast to the inhibitory actions of Brn-3b on several promoters that are activated by Brn-3 alpha. The different actions of the Brn-3 POU factors on the range of nACh receptor genes tested suggests that the novel stimulation of the alpha 2 subunit by Brn-3b is specific to this subunit and not a general feature of nACh receptor genes. PMID- 8713068 TI - Interaction of dibutyltin-3-hydroxyflavone bromide with the 16 kDa proteolipid indicates the disposition of proton translocation sites of the vacuolar ATPase. AB - The organotin complex dibutyltin-3-hydroxyflavone bromide [Bu2Sn(of)Br] has been shown to bind to the 16 kDa proteolipid of Nephrops norvegicus, either in the form of the native protein or after heterologous expression in Saccharomyces and assembly into a hybrid vacuolar H(+)-ATPase. Titration of Bu2Sn(of)Br against the 16 kDa proteolipid results in a marked fluorescence enhancement, consistent with binding to a single affinity site on the protein. Vacuolar ATPase-dependent ATP hydrolysis was also inhibited by Bu2Sn(of)Br, with the inhibition constant correlating well with dissociation constants determined for binding of Bu2Sn(of)Br complex to the proteolipid. The fluorescence enhancement produced by interaction of probe with proteolipid can be back-titrated by dicyclohexylcarbodiimide (DCCD), which covalently modifies Glu140 on helix-4 of the polypeptide. Expression of a mutant proteolipid in which Glu140 was changed to a glycine resulted in assembly of a vacuolar ATPase which was inactive in proton pumping and which had reduced ATPase activity. Co-expression studies with this mutant and wild-type proteolipids suggest that proton pumping can only occur in a vacuolar ATPase containing exclusively wild-type proteolipid. The fluorescent enhancement of affinity of Bu2Sn(of)Br for the mutant proteolipid was not significantly altered, with the organotin complex having no effect on residual ATPase activity. Interaction of the probe with mutant proteolipid was unaffected by DCCD. These data suggest an overlap in the binding sites of organotin and DCCD, and have implications for the organization and structure of proton-translocating pathways in the facuolar H(+)-ATPase. PMID- 8713069 TI - Labelling the Ca(2+)-ATPase of skeletal-muscle sarcoplasmic reticulum with the cross-linker o-phthalaldehyde. AB - The Ca(2+)-ATPase in the sarcoplasmic reticulum of skeletal muscle reacts with o phthalaldehyde (OPA) to form a fluorescent isoindole product. The stoichiometry of labelling of the ATPase is 9 nmol of isoindole/mg of ATPase, corresponding to a 1:1 molar ratio of isoindole: ATPase. There is no evidence for any intermolecular cross-linking. Isoindole formation is faster in the presence of methylamine, but the stoichiometry of labelling is unchanged, whereas in the presence of 2-mercaptoethanol the level of labelling is much higher. It is concluded that OPA reacts with a single Cys residue (defining the specificity of the reaction) in a fast step, subsequent reaction with a Lys residue to form the isoindole being rate-controlling. Labelling the ATPase with OPA in the absence of methylamine leads to total loss of ATPase activity, whereas in the presence of methylamine, the decrease in ATPase activity on reaction is small. We conclude that the loss of ATPase activity probably follows from formation of the intramolecular cross-link rather than from the initial modification of the Cys residue. Reaction with OPA is not affected by the presence of ATP, ADP or Ca2+, so that the reactive Cys is not part of a ligand-binding site. The fluorescence emission spectrum of the labelled ATPase indicates a hydrophobic environment for the isoindole ring. PMID- 8713070 TI - The mechanism of inhibition of the Ca(2+)-ATPase of skeletal-muscle sarcoplasmic reticulum by the cross-linker o-phthalaldehyde. AB - Labelling the Ca(2+)-ATPase of skeletal-muscle sarcoplasmic reticulum with o phthalaldehyde (OPA) results in loss of ATPase activity at a 1:1 molar ration of label to ATPase. The affinity of the ATPase for CA2+ is unaffected, as is the E1/E2 equilibrium constant. The rate of dissociation of Ca2+ from the Ca(2+) bound ATPase is also unaffected and Mg2+ increases the rate of dissociation, as for the unlabelled ATPase. Effects of Mg2+ on the fluorescence intensity of the ATPase labelled with 4-(bromo-methyl)-6,7-dimethoxycoumarin are also unaffected by labelling with OPA, consistent with the fluorescence change reporting on Mg2+ binding at the gating site on the ATPase. The affinity of the ATPase for ATP is reduced by labelling, as is the rate of phosphorylation. The rate of phosphorylation is independent of the concentration of ATP above 25 microM ATP, so that the slow step is the first-order rate constant for phosphorylation by bound ATP. The rate of the back reaction between phosphorylated ATPase and ADP is little affected, suggesting that the slow step in phosphorylation could be the slow conformation step before phosphoryl transfer. The rate of dephosphorylation of the phosphorylated ATPase is also decreased, suggesting that a similar conformation change could be involved in the dephosphorylation step. The rate of the Ca(2+)-transport step appears to be unaffected by labelling. The net result of these changes is that the labelled ATPase is present predominantly in a Ca(2+) free, phosphorylated form at steady state in the presence of ATP. PMID- 8713071 TI - Magnetic and optical properties of copper-substituted alcohol dehydrogenase: a bisthiolate copper (II) complex. AB - Replacement of the catalytic Zn(II) in horse liver alcohol dehydrogenase (HLADH) with copper produces a mononuclear Cu(II) chromophore with a ligand set consisting of two cysteine sulphurs, one histidine nitrogen plus one further atom. The fourth ligand to the metal ion and the conformation of the protein may be altered by addition of exogenous ligands and/or the cofactor NADH. Absorbance, CD, low-temperature magnetic CD (MCD) and EPR spectra are presented of copper substituted HLADH samples in both 'open' and 'closed' conformations and in the presence and absence of the exogenous ligands pyrazole and DMSO. The EPR spectra indicate a strong, predominantly axial field about the copper(II) ion with high copper-thiol (cysteine) covalence. The optical and MCD spectra are interpreted in terms of four d-d transitions to low energy, also reflecting the axial ligand field, and four charge-transfer transitions to copper(II) between 30000 and 16000 cm-1 arising from the two cysteine sulphur atoms which give two pairs of oppositely signed MCD C-terms. These transitions are polarized mainly in the axial plane defined by Cys-46, Cys-174 and His-67. The binary complex formed with pyrazole displays quite different EPR and optical spectra which can be understood in terms of a rotation of the copper hole-orbital away from the axial plane thus decreasing sharply the copper-thiol covalence. The magneto-optical spectra in the presence and absence of DMSO are indistinguishable. PMID- 8713072 TI - Functional characterization of a recombinant sodium-dependent nucleoside transporter with selectivity for pyrimidine nucleosides (cNT1rat) by transient expression in cultured mammalian cells. AB - We have demonstrated that monkey kidney (COS-1) cells have a single type of nucleoside transport process, which, because it was equilibrative, sodium independent and could be inhibited by nitrobenzylthioinosine (NBMPR), was identified as the 'equilibrative sensitive' or 'es' transporter. Using NBMPR or dilazep to inhibit the endogenous nucleoside transport activity, we have transiently expressed a cDNA that encodes an inhibitor-insensitive, concentrative nucleoside transporter protein (cNT1rat) of rat intestine in COS-1 cells. The production of recombinant cNT1rat was examined by immunoblotting using an epitope tagged construct and by analysis of inward fluxes of 3H-labelled nucleosides. Recombinant cNT1rat was sodium-dependent and selective for pyrimidine nucleosides, with approximately Km values of 21 microM, 12.5 microM and 15 microM for uridine, thymidine and adenosine, respectively. Although adenosine exhibited high affinity for the recombinant transporter, its Vmax value was low. A variety of anti-viral and anti-cancer nucleoside drugs inhibited cNT1rat-mediated uptake of uridine by transfected COS-1 cells although to different extents (Floxidine > Idoxuridine > Zidovudine > Zalcitabine > Cytarabine > Gemcitabine), suggesting that the concentrative pyrimidine-selective nucleoside transporters, of which cNT1rat is a representative, may play a role in cellular uptake of these drugs. The cNT1rat/COS-1 expression system is a useful tool for analysis of cNT1rat mediated transport processes. PMID- 8713073 TI - Evidence that residues exposed on the three-fold channels have active roles in the mechanism of ferritin iron incorporation. AB - Iron is thought to enter the ferritin cavity via the three-fold channel, which is lined in its narrowest part by the residues Asp-131 and Glu-134. We describe here variants of human ferritins with active and inactive ferroxidase centres having Asp-131 and Glu-134 substituted with Ala and Ala or with Ile and Phe respectively. The two types of substitution had similar effects on ferritin functionality: (i) they decreased the amount of iron incorporated from Fe(II) solutions and decreased ferroxidase activity by about 50%; (ii) they inhibited iron incorporation from Fe(III) citrate in the presence of ascorbate; (iii) they resulted in loss of Fe and Tb binding sites; and (iv) they resulted in a marked decrease in the inhibition of iron oxidation by Tb (but not by Zn). In addition, it was found that substitution with Ala of Cys-130 and His-118, both of which face the three-fold channel, decreased the capacity of H-ferritin to bind terbium and to incorporate iron from Fe(III) citrate in the presence of ascorbate. The results indicate that: (i) in three-fold channels are the major sites of iron transfer into the cavity of H- and L-ferritins; (ii) at least two metal binding sites are located on the channels which play an active role in capturing and transferring iron into the cavity; and (iii) the permeability of the channel is apparently not affected by the hydrophilicity of its narrowest part. In addition, it is proposed that iron incorporation from Fe(III) citrate complexes in the presence of ascorbate is a reliable, and possibly more physiological, approach to the study of ferritin functionality. PMID- 8713074 TI - Synergistic activation of PtdIns 3-kinase by tyrosine-phosphorylated peptide and beta gamma-subunits of GTP-binding proteins. AB - Stimulation of differentiated THP-1 cells by insulin led to rapid accumulation of PtdIns(3,4,5)P3, a product of PtdIns 3-kinase. Stimulation of the GTP-binding protein-linked receptor by N-formylmethionyl-leucyl-phenylalanine (fMLP) also induced the accumulation of PtdIns(3,4,5)P3 in the cells. The effect of insulin was, while that of fMLP was not, accompanied by increased PtdIns 3-kinase activity in the anti-phosphotyrosine immuno-precipitate. The combination of insulin and fMLP induced more PtdIns(3,4,5)P3 production than the sum of the individual effects. The insulin-induced recruitment of PtdIns 3-kinase activity in the anti-phosphotyrosine immunoprecipitate was unaffected by the combined treatment with fMLP. To investigate the mechanism underlying the synergistic accumulation of PtdIns(3,4,5)P3, we separated the cytosolic proteins of THP-1 cells on a Mono Q column. PtdIns 3-kinase activities were eluted in two peaks, and one of the peaks markedly increased on the addition of beta gamma-subunits of GTP-binding proteins (G beta gamma). The other peak was affected only slightly by G beta gamma, but was synergistically increased by G beta gamma and a tyrosine phosphorylated peptide which was synthesized accordingly to the amino acid sequence of insulin receptor substrate-1. The activity in the latter fraction was completely immunoprecipitated by an antibody against the regulatory subunit of PtdIns 3-kinase (p85). These results suggest that the conventional PtdIns 3 kinase (p85/p110), which has been implicated in insulin-induced cellular events, or a closely related isoenzyme is controlled by a combination of a tyrosine phosphorylated protein and a GTP-binding protein in intact cells. PMID- 8713075 TI - Genetic analysis of the phenobarbital regulation of the cytochrome P-450 2b-9 and aldehyde dehydrogenase type 2 mRNAs in mouse liver. AB - The aim of this study was to investigate the effect of the genetic background on the phenobarbital inducibility of cytochrome P-450 2b-9, cytochrome P-450 2b-10 and aldehyde dehydrogenase type 2 mRNAs in mice. We analysed the basal expression and the phenobarbital inducibility of both cytochrome P-450 mRNAs by semi quantitative specific reverse transcription-PCR analyses in five inbred mouse strains (A/J,BALB/cByJ,C57BL/6J, DBA/2J and SWR/J). Male mice constitutively expressed cytochrome P-450 2b-9 and cytochrome P-450 2b-10 mRNAs, but a number of differences in their response to phenobarbital were observed. In all these mouse strains, phenobarbital induced cytochrome P-450 2b-10 mRNA whereas it could have either a positive or a negative effect on cytochrome P-450 2b-9 expression, depending on the strain and the sex of the mice. Specifically, phenobarbital increased cytochrome P-450 2b-9 expression in C57BL/6J males while it decreased it in DBA/2J mice. Interestingly, dexamethasone was able to mimic the phenobarbital effect on both cytochromes P-450 in these two strains. Aldehyde dehydrogenase type 2 mRNA was always induced by phenobarbital, except in the C57BL/6J strain. Genetic analysis revealed that the phenobarbital-inducible phenotype was either a semi-dominant or a recessive trait in F1 animals from a C57BL/6J x DBA/2J cross for the cytochrome P-450 2b-9 and the aldehyde dehydrogenase type 2 genes, respectively. This study suggests that the genetic basis for phenobarbital induction in mice depends on the target gene, and that more than one regulatory step would by involved in this response pathway. PMID- 8713076 TI - Characterization of the effects of Ca2+ depletion on the synthesis, phosphorylation and secretion of caseins in lactating mammary epithelial cells. AB - We have examined the effects of depleting lumenal Ca2+ on the synthesis, phosphorylation and secretion of caseins in lactating mouse mammary cells by using inhibitors of the endoplasmic reticulum Ca(2+)-ATPase or the ionophore ionomycin in the absence of external Ca2+. Treatment with these drugs resulted in a transient increase in the cytosolic Ca2+ concentration due to Ca2+ mobilization. Protein synthesis over a 1 h period was substantially inhibited by Ca2+ depletion, but in a pulse-chase protocol secretion of pre-synthesized proteins was unaffected by Ca2+ depletion. Analysis of polysome profiles showed that Ca2+ depletion resulted in a loss of polysomes, consistent with an inhibition of initiation of protein synthesis. Neither treatment with Ca(2+) ATPase inhibitors to deplete endoplasmic reticulum Ca2+ nor treatment with ionomycin/EGTA had any effect on an early phase of phosphorylation of alpha- or beta/gamma-caseins, but Ca2+ depletion resulted in a decrease in a late phase of casein phosphorylation. These results indicate that lumenal Ca2+ is required to maintain protein synthesis in lactating mammary cells but is not required for protein secretion, and that Ca2+ accumulation in the Golgi cisternae is required for a late but not for an early phase of casein phosphorylation. PMID- 8713077 TI - Evidence for the presence of essential histidine and cysteine residues in platelet cGMP-inhibited phosphodiesterase. AB - Camp is a major regulator of platelet function. cGMP-inhibited phosphodiesterase (cGI-PDE) is the predominant platelet enzyme hydrolysing cAMP. The pH-rate profile plot for this enzyme yields pKa values of 6.5 and 9.0, consistent with histidine and cysteine residues respectively. Diethyl pyrocarbonate (DEP) inactivates cGI-PDE in a time- and concentration-dependent manner, and this effect was rapidly reversed by hydroxylamine. It was estimated that 2 mol of histidine residues per mol of enzyme were responsible for the loss of catalytic activity, as deduced from the correlation of the difference spectrum at 240 nm of the DEP-modified cGI-PDE with the enzyme activity. N-Ethylmaleimide (NEM) and 5.5'-dithiobis-(2-nitrobenzoic acid) (DTNB) inactivate cGI-PDE in a time- and concentration-dependent manner, suggesting the selective modification of a cysteine residue. AMP protects the enzyme against DEP, NEM and DTNB, suggesting the presence of histidine and cysteine residues at the active site of cGI-PDE. [14C]DEP incorporation in the presence of AMP or cGMP indicates the protection of two histidine residues by each nucleotide. These residues are different for each agent, since the combination of AMP and cGMP protects four histidine residues. [3H]NEM incorporation showed that 1 mol of cysteine per mol of cGI-PDE was protected by AMP, but not only by cGMP. We conclude that cGI-PDE possesses two essential histidine residues for activity, two additional histidines for cGMP inhibition, and one cysteine residue at the active site. PMID- 8713078 TI - Effect of multiple cytokines plus bacterial endotoxin on glucose and nitric oxide production by cultured hepatocytes. AB - Treatment of cultured hepatocytes with a combination of cytokines, including tumour necrosis factor-alpha, interferon-gamma and interleukin-1 beta, plus lipopolysaccharide resulted in a time-dependent induction of nitric oxide (NO) synthase (as measured by NO2- (+) NO3- production) and inhibition of hepatic gluconeogenesis and glycogen breakdown. The inhibition of glucose release was comparable with the observed following treatment of rats with lipopolysaccharide or treatment of isolated hepatocytes with artificial NO donors. In addition, this effect was also evident with all substrates tested that enter the gluconeogenic pathway below the level of phosphoenolpyruvate carboxykinase, suggesting that this combination of cytokines may underlie the inhibition of gluconeogenesis observed in endotoxic shock. The maximal inhibition of glucose output required the presence of all the cytokines plus lipopolysaccharide, whereas the induction of NO synthase was independent of the lipopolysaccharide when the cytokines were employed. Inclusion of interferon-gamma was essential to obtain a maximal response for either parameter. Inclusion of 1 mM N(G)-monomethyl-L-arginine in the incubation abolished the increase in NO2- (+) NO3- observed with the complete cytokine mixture and various combinations; however, it failed to prevent the inhibition in glucose output, indicating that mechanisms other than NO underlie the cytokine-induced inhibition of glucose release. PMID- 8713079 TI - Characterization of the stilbenedisulphonate binding site on band 3 Memphis variant II (Pro-854-->Leu). AB - Band 3 Memphis variant II is a mutant anion-exchange protein associated with the Diego a+ blood group antigen. There are two mutations in this transporter: Lys-56 ->Glu within the cytoplasmic domain, and Pro-854-->Leu within the membrane-bound domain. The Pro-854 mutation, which is thought to give rise to the antigenicity, is located within the C-terminal subdomain of the membrane-bound domain. Yet, there is an apparent enhancement in the rate of covalent binding of H2DIDS (4,4' di-isothiocyanatodihydro-2, 2'-stilbenedisulphonate) to 'lysine A' (Lys-539) in the N-terminal subdomain, suggesting widespread conformational changes. In this report, we have used various kinetic assays which differentiate between conformational changes in the two subdomains, to characterize the stilbenedisulphonate site on band 3 Memphis variant II. We have found a significantly higher H2DIDS (a C-terminal-sensitive inhibitor) affinity for band 3 Memphis variant II, due to a lower H2DIDS 'off' rate constant, but no difference was found between mutant and control when DBDS (4,4'-dibenzamido-2,2' stilbenedisulphonate) (a C-terminal-insensitive inhibitor) 'off' rates were measured. Furthermore, there were no differences in the rates of covalent binding to lysine A, for either DIDS (4,4'-di-isothiocyanato-2,2'-stilbenedisulphonate) or H2DIDS. However, the rate of covalent intrasubunit cross-linking of Lys-539 and Lys-851 by H2DIDS was abnormally low for band 3 Memphis variant II. These results suggest that the Pro-854-->Leu mutation causes a localized conformational change in the C-terminal subdomain of band 3. PMID- 8713080 TI - Effects of steroids and verapamil on P-glycoprotein ATPase activity: progesterone, desoxycorticosterone, corticosterone and verapamil are mutually non exclusive modulators. AB - P-glycoprotein (P-gp) is a membranous ATPase responsible for the multidrug resistance (MDR) phenotype. Using membrane vesicles prepared from the highly resistant cell line DC-3F/ADX we studied the influence of P-gp ATPase activity of four progesterone derivatives which specifically bind to P-gp and reverse MDR. Progesterone and desoxycorticosterone stimulate P-gp ATPase activity with, respectively, apparent concentrations giving half-maximal activation of 20-25 microM and 40-50 microM, and activation factors of 2.3 (at 100 microM progesterone) and 1.8 (at 170 microM desoxycorticosterone). Hydrocortisone above 100 microM stimulates P-gp ATPase activity while corticosterone has no apparent stimulating effect. Our data are consistent with the location of the binding sites for the progesterone derivatives on the P-gp membranous domain. The effects of these steroids on verapamil-stimulated P-gp ATPase activity support a non competitive mechanism, i.e. the binding sites for verapamil and steroids are mutually non-exclusive for P-gp ATPase modulation. A similar non-competitive inhibition of progesterone-stimulated P-gp ATPase activity by desoxycorticosterone or by corticosterone leads to the conclusion that these steroids, although sharing related structures, have distinct modulating sites on P-gp. As expected from their mutually non-exclusive interactions on P-gp, progesterone and verapamil when mixed induce a synergistic modulation of P-gp ATPase activity. Since drug transport by P-gp is believed to be coupled to its ATPase activity, a corresponding synergistic effect of these two modulators for the inhibition of P-gp-mediated drug resistance can be expected. PMID- 8713081 TI - Affinity for the nuclear compartment and expression during cell differentiation implicate phosphorylated Groucho/TLE1 forms of higher molecular mass in nuclear functions. AB - The Drosophila protein Groucho is involved in embryonic segmentation and neural development, and is implicated in the Notch signal transduction pathway. We are investigating the molecular mechanisms underlying the function of Groucho and of its mammalian homologues, the TLE ('transducin-like Enhancer of split') proteins. We show that Groucho/TLE1 proteins are phosphorylated. We also show that two populations of phosphorylated Groucho proteins can be identified based on their interaction with the nuclear compartment. More slowly migrating proteins with an apparent molecular mass of roughly 110 kDa interact strongly with the nuclei, while faster migrating proteins displaying molecular masses roughly 84-85 kDa show lower affinity for the nuclear compartment. Similarly, TLE1 proteins with an apparent molecular mass of roughly 118 kDa exhibit higher affinity for the nuclear compartment than do faster migrating forms with apparent molecular masses of 90-93 kDa. Moreover, we show that the nuclear, more slowly migrating, TLE1 proteins are induced during neural determination of P19 embryonic carcinoma cells. These results implicate phosphorylation in the activity of Groucho/TLE1 proteins and suggest that phosphorylated forms of higher molecular mass are involved in nuclear functions. Finally, we show that different TLE proteins respond in different ways to the neural commitment of P19 cells, suggesting that individual members of this protein family may have non-redundant functions. PMID- 8713082 TI - An uncleaved glycosylphosphatidylinositol signal mediates Ca(2+)-sensitive protein degradation. AB - Inv-gp80 is a chimeric protein which contains a signal for the attachment of a glycosylphosphatidylinositol (GPI) anchor. When expressed in Dictyostelium discoideum, this protein fails to become GPI anchored and is retained within the cell as an integral membrane protein. We have compared the subcellular localization and degradation of Inv-gp80 with that of its intracellular but soluble counterpart, Inv-gp80sc. Inv-gp80sc lacks the hydrophobic C-terminal 22 amino acids of Inv-gp80. The N-linked oligosaccharides of both Inv-gp80 and Inv gp80sc remained sensitive to endoglycosidase H, and both proteins co-fractionated with endoplasmic reticulum marker enzymes on Percoll gradients. Under normal conditions, Inv-gp80 displayed a half-life (t 1/2) of 90 min, while Inv-gp80sc displayed a t 1/2 of 120 min. The degradation of both proteins required ATP, was inhibited by tosyl phenylalanylchloromethane (Tos-Phe-CH2Cl) and was insensitive to inhibitors of lysosomal function. While depletion of Ca2+ from the endoplasmic reticulum had no effect on the degradation of Inv-gp80sc, it stimulated the degradation of Inv-gp80. When the GPI anchor signal sequence of Inv-gp80 was replaced with the transmembrane domain of the interleukin-2 receptor, the degradation of the protein was no longer influenced by Ca2+ fluxes. The data suggest that while the GPI anchor sequence of Inv-gp80 does not contain determinants regulating the degradation of the protein under basal conditions, it targets Inv-gp80 for rapid degradation under conditions where Ca2+ is depleted from the endoplasmic reticulum. PMID- 8713083 TI - Characterization of a platelet-activating factor acetylhydrolase secreted by the nematode parasite Nippostrongylus brasiliensis. AB - Nippostrongylus brasiliensis, a small nematode parasite of the gastrointestinal tract of rodents, secretes an enzyme that cleaves the proinflammatory molecule platelet-activating factor to its inactive lyso-form. The enzyme activity of Ca(2+)-dependent and does not exhibit interfacial activation. It does not require the addition of reducing agents for maximal activity, and is not inhibited by thiol-active reagents. Sensitivity of inhibitors suggests the involvement of serine and histidine residues in the enzyme activity. As described for other platelet-activating factor acetylhydrolases, it cannot cleave, nor is it inhibited by, long-chain diacyl phospholipids that are typical substrates for phospholipases A2. The purified enzyme was resolved by SDS/PAGE as a heterodimer composed of two protein subunits with apparent molecular masses of 38 and 25 kDa. The properties of the nematode enzyme thus differ from those described for the mammalian enzymes, but are more closely related to those of an acetylhydrolase than a phospholipase. PMID- 8713084 TI - Secondary structure of NADPH: protochlorophyllide oxidoreductase examined by circular dichroism and prediction methods. AB - To study the secondary structure of the enzyme NADPH: protochlorophyllide oxidoreductase (PCOR), a novel method of enzyme isolation was developed. The detergent isotridecyl poly-(ethylene glycol) ether (Genapol X-080) selectively solubilizes the enzyme from a prolamellar-body fraction isolated from wheat (Triticum aestivum L.). The solubilized fraction was further purified by ion exchange chromatography. The isolated enzyme was studied by fluorescence spectroscopy at 77 K, and by CD spectroscopy. The fluorescence-emission spectra revealed that the binding properties of the substrate and co-substrate were preserved and that photo-reduction occurred. The CD spectra of PCOR were analysed for the relative amounts of the secondary structures, alpha-helix, beta-sheet, turn and random coil. The secondary structure composition was estimated to be 33% alpha-helix, 19% beta-sheet, 20% turn and 28% random coil. These values are in agreement with those predicted by the Predict Heidelberg Deutschland and self optimized prediction method from alignments methods. The enzyme has some amino acid identity with other NADPH-binding enzymes containing the Rossmann fold. The Rossmann-fold fingerprint motif is localized in the N-terminal region and at the expected positions in the predicted secondary structure. It is suggested that PCOR is anchored to the interfacial region of the membrane by either a beta-sheet or an alpha-helical region containing tryptophan residues. A hydrophobic loop region could also be involved in membrane anchoring. PMID- 8713085 TI - Structural investigation of the molybdenum site of the periplasmic nitrate reductase from Thiosphaera pantotropha by X-ray absorption spectroscopy. AB - The molybdenum centre of the periplasmic respiratory nitrate reductase from the denitrifying bacterium Thiosphaera pantotropha has been probed using molybdenum K edge X-ray absorption spectroscopy. The optimum fit of the Mo(VI) EXAFS suggests two ==O, three -S- and either a fourth -S- or an -O-/-N- as molybdenum ligands in the ferricyanide-oxidized enzyme. Three of the -S- ligands are proposed to be the two sulphur atoms of the molybdopterin dithiolene group and Cys-181. Comparison of the EXAFS of the ferricyanide-oxidized enzyme with that of a nitrate-treated sample containing 30% Mo(V) suggests that the Mo(VI)-->Mo(V) reduction is accompanied by conversion of one ==O to -O-. The best fit to the Mo(IV) EXAFS of dithionite-reduced enzyme was obtained using one ==O, one -O- and four -S-/-Cl ligands. The periplasmic nitrate reductase molybdenum co-ordination environment in both the Mo(VI) and Mo(IV) oxidation states is distinct from that found in the membrane-bound respiratory nitrate reductase. PMID- 8713086 TI - Cyclophilin B mediates cyclosporin A incorporation in human blood T-lymphocytes through the specific binding of complexed drug to the cell surface. AB - Cyclophilin B (CyPB) is a cyclosporin A (CsA)-binding protein located within intracellular vesicles and released in biological fluids. We recently reported the specific binding of this protein to T-cell surface receptor which is internalized even in the presence of CsA. These results suggest that CyPB might target the drug to lymphocytes and consequently modify its activity. To verify this hypothesis, we have first investigated the binding capacity and internalization of the CsA-CyPB complex in human peripheral blood T-lymphocytes and secondly compared the inhibitory effect of both free and CyPB-complexed CsA on the CD3-induced activation and proliferation of T-cells. Here, we present evidence that both the CsA-CyPB complex and free CyPB bind to the T-lymphocyte surface, with similar values of Kd and number of sites. At 37 degrees C, the complex is internalized but, in contrast to the protein, the drug is accumulated within the cell. Moreover, CyPB receptors are internalized together with the ligand and rapidly recycled to the cell surface. Finally, we demonstrate that CyPB-complexed CsA remains as efficient as uncomplexed CsA and that CyPB enhances the immunosuppressive activity of the drug. Taken together, our results support the hypothesis that surface CyPB receptors may be related to the selective and variable action of CsA, through specific binding and targeting of the CyPB-CsA complex to peripheral blood T-lymphocytes. PMID- 8713087 TI - Involvement of the N-terminal part of cyclophilin B in the interaction with specific Jurkat T-cell binding sites. AB - Cyclophilin B (CyPB) is secreted in biological fluids such as blood or milk and binds to a specific receptor present on the human lymphoblastic cell line Jurkat and on human peripheral blood lymphocytes. This study was intended to specify the areas of CyPB that are involved in the interaction with the receptor. A synthetic peptide corresponding to the first 24 N-terminal amino acid residues of CyPB was shown to specifically recognize the receptor. Moreover, modification of Arg18 of CyPB by p-hydroxyphenlglyoxal led to a dramatic loss of affinity for the receptor. However, when this residue was replaced by an alanine residue using site-directed mutagenesis, no modification of the binding properties was found, suggesting that Arg18 is not directly involved but is sufficiently close to the interaction site to interfere with the binding when modified. Competitive binding experiments using a chimaeric protein made up of the 24 N-terminal amino acid residues of CyPB fused to the cyclophilin A core sequence confirmed the involvement of this region of CyPB in receptor binding. PMID- 8713088 TI - Molecular recognition of peptide and non-peptide ligands by the extracellular domains of neurohypophysial hormone receptors. AB - This study was designed to ascertain whether the extracellular loops of vasopressin/oxytocin receptors bind ligands and, if so, to locate the molecular determinants of this ligand-receptor interaction. Ligand-binding studies were employed using a rat liver V1a vasopressin receptor preparation and both peptide and non-peptide receptor ligands. Synthetic peptides corresponding to defined regions of the extracellular surface of the neurohypophysial hormone receptors recognized radioligands. These receptor mimetics inhibited the binding of radioligands to the V1a receptor with apparent affinities (pKi) ranging from 3.1 to 6.75. The same mimetics had no effects on the binding of angiotensin II to the rat AT1 receptor, indicating specificity for V1a receptor ligands. A mimetic peptide (DITYRFRGPDWL) of the first extracellular loop (ECII) of the V1a vasopressin receptor also inhibited vasopressin-stimulated, but not angiotensin II-stimulated, glycogen phosphorylase in isolated rat hepatocytes. In contrast, scrambled ECII mimetics displayed greatly reduced affinity for vasopressin. In addition, the role of peptide side-chain versus main-chain atoms in the binding of ligands by vasopressin receptors was addressed using retro-inverso peptide mimetics. Our findings indicate a precise orientation of the extracellular receptor surface (particularly the ECII domain) which facilitates the initial 'capture' of both peptide and non-peptide ligands. Moreover, the data indicate that the main-chain atoms of both a major binding-site determinant in the first extracellular loop of the receptor and the neurohypophysial hormones contribute significantly to the ligand-receptor interaction. These findings also suggest that soluble receptor-binding domains have therapeutic potential. PMID- 8713089 TI - Activation of protein kinase C by lysophosphatidic acid: dependence on composition of phospholipid vesicles. AB - Lysophosphatidic acid (LPA) has attracted recent attention as a major serum derived regulator implicated in responses to vascular injury and inflammation, in tumour invasiveness and in neuronal signalling and remodelling. Although the possibility of a specific G-protein-coupled LPA receptor protein has been suggested, characterization of such a receptor is lacking. Since LPA can activate protein kinase C (PKC) pathways in many cells and PKC activators mimic many LPA effects, the possibility of more direct LPA effects on PKC was investigated. Phosphatidylcholine (PC)/phosphatidylserine (PS)/diacylglycerol (DAG) lipid vesicles of defined acyl chain composition were used to activate the enzyme. At total concentrations of saturated PC/PS + DAG vesicles (2-3 mM) that provided maximal PKC activation, 1-10 mol % [18:1]-LPA led to a further approx. 2-fold activation of PKC alpha. At lower lipid concentrations, a greater increase was observed with LPA concentrations up to 16-20 mol %. Higher concentrations of LPA were inhibitory. The LPA activation of PKC was dependent on the presence of DAG, PS and Ca2+. [18:1]-Lysophosphatidylcholine produced similar PKC activation in PC/PS/DAG vesicles. [14:0]-LPA was less effective, and longer-chain saturated lysolipids were ineffective. In unsaturated PC/PS vesicles, very little to no effect of LPA was discernable. These results suggest that physiologically or pathologically relevant concentrations of LPA can contribute to PKC activation depending on the composition of the lipid membrane. We hypothesize that LPA may affect the formation of lipid domains that are recognized by the enzyme. PMID- 8713090 TI - Synthesis of non-hydroxy-galactosylceramides and galactosyldiglycerides by hydroxy-ceramide galactosyltransferase. AB - Galactosylceramide (GalCer) is the major glycolipid in brain. In order to characterize the activity of brain UDPgalactose: ceramide galactosyltransferase (CGalT), it has been stably expressed in CGalT-negative Chinese hamster ovary (CHO) cells. After fractionation of transfected cells, CHO-CGT, on sucrose gradients, the activity resides at the density of endoplasmic reticulum and not of Golgi. A lipid chromatogram from CHO-CGT cells revealed two new iodine staining spots identified as GalCer, since they comigrate with GalCer standards, can be metabolically labelled with [3H]galactose, are recognized by anti-GalCer antibodies, and are resistant to alkaline hydrolysis. A third [3H]galactose lipid was identified as galactosyldiglyceride. In the homogenate CGalT displays a 25 fold preference for hydroxy fatty acid-containing ceramides. Remarkably, endogenous GalCer of transfected cells contains exclusively non-hydroxy fatty acids: fast atom bombardment and collision-induced dissociation mass spectrometric analysis revealed mainly C16:0 in the lower GalCer band on TLC and mainly C22:0 and C24:0 in the upper band. Our results suggest that CGalT galactosylates both hydroxy- and non-hydroxy fatty acid-containing ceramides and diglycerides, depending on their local availability. Thus, CGalT alone may be responsible for the synthesis of hydroxy- and non-hydroxy-GalCer, and galactosyldiglyceride in myelin. PMID- 8713091 TI - Molecular characterization of quail apolipoprotein very-low-density lipoprotein II: disulphide-bond-mediated dimerization is not essential for inhibition of lipoprotein lipase. AB - As part of the avian reproductive effort, large quantities of triglyceride-rich very-low-density lipoprotein (VLDL) particles are transported by receptor mediated endocytosis into the female germ cells. Although the oocytes are surrounded by a layer of granulosa cells harbouring high levels of active lipoprotein lipase, non-lipolysed VLDL is transported into the yolk. This is because VLDL particles from laying chickens are protected from lipolysis by apolipoprotein (apo)-VLDL-II, a potent dimeric lipoprotein lipase inhibitor [Schneider, Carroll, Severson and Nimpf (1990) J. Lipid Res. 31, 507-513]. To determine whether this protection depends on dimer formation and constitutes a general mechanism to ensure high levels of yolk triglycerides for embryonic utilization in birds, we have now molecularly characterized apo-VLDL-II in the Japanese quail, a frequently used avian species. Quail apo-VLDL-II shows 72% amino acid identity with the chicken protein, with most replacements being in the C-terminal region. Importantly, quail apo-VLDL-II lacks the single cysteine residue present eight residues from the C-terminus of chicken apo-VLDL-II, which is responsible for dimerization of the chicken lipoprotein lipase inhibitor. Nevertheless, monomeric quail and dimeric chicken apo-VLDL-II display, on a molar basis, identical inhibitory effects on lipoprotein lipase, underscoring the biological importance of their function. Furthermore secondary structure prediction of the 3'-untranslated region of the quail message supports a role for loop structures in the strictly oestrogen-dependent production of the lipoprotein lipase inhibitors. Our findings shed new light on the essential role of this small, hormonally regulated, protein in avian reproduction. PMID- 8713092 TI - Structural changes in subdomain 2 of G-actin observed by fluorescence spectroscopy. AB - The influence of DNase I binding to Ca-ATP-G-actin and of Ca2+/Mg2+ and ATP/ADP exchange on the conformation of G-actin were investigated by measuring the fluorescence of dansyl cadaverine (DC) conjugated to Gln41 in subdomain 2 of the protein. Fluorescence resonance energy transfer (FRET) between this probe and N [4-(dimethylamino)-3,5-dinitrophenyl]maleimide (DDPM) attached to Cys374 in subdomain 1 was also measured. Contrary to an earlier report [dos Remedios, Kiessling and Hambly (1994) in Synchrotron Radiation in the Biosciences (Chance, B., Deisenhofer, J., Ebashi, S., Goodhead, D. T., Helliwell, J. R., Huxley, H. E., Iizuka, T., Kirz, J., Mitsui, T., Rubenstein, E. et al., eds.), pp. 418-425, Oxford University Press, Oxford], the distance between these probes did not change significantly when DNase I was bound to actin. A small but reproducible increase in the quantum yield and a blue shift of the DC fluorescence maximum were observed when bound Ca2+ was replaced by Mg2+. A large increase (about 70%) in the quantum yield and an approx. 12 nm blue shift of the emission spectrum occurred when ATP in Mg-G-actin was replaced by ADP. These changes were not accompanied by any significant change in the FRET distance between the dansyl donor and DDPM acceptor probes. A substantial change in the fluorescence of DC actin was observed after proteolytic removal of the last three residues of actin, in accordance with earlier evidence suggesting that there is a conformational coupling between subdomain 2 and the C-terminal segment in subdomain 1 of actin. The results are discussed in relation to recently published data obtained with another fluorescent probe and to earlier observations based on limited cleavage using proteolytic enzymes. PMID- 8713093 TI - Measurement of the turnover of glycogen phosphorylase by GC/MS using stable isotope derivatives of pyridoxine (vitamin B6). AB - The majority of vitamin B6 in the body is in skeletal muscle, bound as the cofactor pyridoxal 5'-phosphate to one abundant protein, glycogen phosphorylase. Previous work has established that radiolabelled vitamin B6 can be used as a turnover label for glycogen phosphorylase. In this study, a stable isotope derivative of pyridoxine {dideuterated pyridoxine; 3-hydroxy-4-(hydroxymethyl) -5 [hydroxymethyl-2H2]-2-methylpyridine} ([2H2]PN) has been used as a metabolic tracer to study the kinetics of labelling of the body pools of vitamin B6 in mice. A non-invasive method was developed in which the isotope abundance of the urinary excretory product of vitamin B6 metabolism, 4-pyridoxic acid, was analysed by GC/MS. The change in isotope abundance of urinary 4-pyridoxic acid following administration of [2H2]PN reflects the kinetics of labelling of the body pools of vitamin B6, and yields, non-invasively, the rate of degradation of glycogen phosphorylase. PMID- 8713096 TI - Characterization of a Porphyromonas gingivalis gene prtR that encodes an arginine specific thiol proteinase and multiple adhesins. AB - Cysteine proteinases of Porphyromonas gingivalis have been implicated as major virulence factors in the development of periodontitis. Several groups have reported the characterisation of similar genes encoding the same arginine specific thiol proteinase from P. gingivalis; however, the reported size and structure of the genes have varied. We report here the complete nucleotide sequence of the gene prtR that encodes a polyprotein containing the Arg-specific proteinase and multiple haemagglutinins/adhesins. The nascent polyprotein consists of a putative leader sequence and a prosequence followed by the 45 kDa Arg-specific proteinase and 44, 15, 17 and 27 kDa sequence-related adhesins in that order. The size and structure of the prtR are consistent with the size of the mRNA transcript (5.3 kb) and the size and sequences of the individual protein components purified from P. gingivalis. PMID- 8713095 TI - Role of interleukin 6 and corticosteroids in the regulation of expression of glutathione S-transferases in primary cultures of rat hepatocytes. AB - The effect of recombinant interleukin 6 (rIL-6) on the transcript levels of rat glutathione S-transferase (GST) genes rGSTA2, rGSTP1, rGSTM1 and rGSTM2 was examined in primary cultures of rat hepatocytes. rIL-6 had little effect on the increase in expression of rGSTP1 that occurs in cultured hepatocytes. Dexamethasone (DEX), in contrast, prevented the expression of rGSTP1 by hepatocytes, and rIL-6 in combination with DEX had no additional effect. Neither rIL-6 nor DEX alone had a significant effect on the transcript levels of rGSTA2, rGSTM1 and rGSTM2 in cultured hepatocytes. However, when both were present (15 ng/ml rIL-6 and 10(-7) M DEX) the transcript levels of rGSTA2, rGSTM1 and rGSTM2 decreased significantly (P < 0.05) after 48 h in culture. If the rIL-6 was removed from the cultures after 24 h, the levels of transcripts recovered and were the same at 48 h as cells cultured without rIL-6 for the entire period. Dose response relationships of rIL-6 with 10(-7) M DEX were determined for transcripts of each GST isoenzyme and the IC50 values were between 1.5 and 7.5 ng/ml. Declines in transcript levels of rGSTA2 were observed with rIL-6 plus 10(-8) or 10(-7) M DEX but not with rIL-6 plus 10(-9), 10(-6), or 10(-5) M DEX. To determine if the cytokine and glucocorticoid effects were mediated by sequences in the 5'-flanking sequence of rGSTA2, a plasmid construct containing a 1.6 kb fragment of the 5'-flanking sequence of the rGSTA2 gene and the chloramphenicol acetyltransferase (CAT) reporter gene was used to transfect rat hepatocytes in primary culture. The addition of rIL-6 and DEX to the culture medium caused a significant (P < 0.05) decrease in CAT activity after 48 h in culture. If rIL-6 was removed after 24 h in culture, CAT activity after an additional 24 h in culture was greater than the CAT activity in cells cultured for 48 h without rIL 6. Therefore cytokines and glucocorticoids may be important physiological regulators of GST expression. PMID- 8713094 TI - 11 beta-hydroxysteroid dehydrogenase type 1 expression in 2S FAZA hepatoma cells is hormonally regulated: a model system for the study of hepatic glucocorticoid metabolism. AB - 11 beta-Hydroxysteroid dehydrogenase (11 beta-HSD) is a key enzyme in glucocorticoid metabolism, catalysing the conversion of active glucocorticoids into their inactive 11-keto metabolites, thus regulating glucocorticoid access to intracellular receptors. The type 1 isoform (11 beta-HSD 1) (EC 1.1.1.146) is widely distributed, with particularly high levels in liver, where accumulating evidence suggests that it acts as an 11 beta-reductase, regenerating active glucocorticoids. Investigation of the function and regulation of 11 beta-HSD 1 in liver has been hampered by the lack of hepatic cell lines which express 11 beta HSD 1. Here, we describe 11 beta-HSD 1 mRNA expression and activity in 2S FAZA cells, a continuously cultured rat liver cell line. In intact 2S FAZA cells 11 beta-HSD 1 acts predominantly as a reductase, with very low dehydrogenase activity. In 2S FAZA cells 11 beta-HSD 1 activity and mRNA expression are regulated by hormones, with dexamethasone increasing activity and insulin, forskolin and insulin-like growth factor 1 decreasing it. Transfection of 2S FAZA cells with a luciferase reporter gene driven by the proximal promoter of the rat 11 beta-HSD 1 gene demonstrates that sequences which can mediate the responses to insulin, dexamethasone and forskolin all lie within 1800 bp of the transcription start site. PMID- 8713097 TI - Nitrite reductase from Desulfovibrio desulfuricans (ATCC 27774)--a heterooligomer heme protein with sulfite reductase activity. AB - The membrane bound cytochrome c nitrite reductase from the sulfate reducer Desulfovibrio desulfuricans (ATCC 27774) was found to have a high specific activity in the reduction of sulfite, producing stoichiometric amounts of sulfide. The K(m) for sulfite in the MV+.:sulfite oxidoreductase assay is 0.75 mM, and the specific activity 2.06 mumolH2/min/mg. Visible and EPR spectroscopies studies indicate that the enzyme high-spin heme reacts with sulfite in the oxidised state, and that sulfide partially reduces the enzyme. The redoxcycled enzyme, using H2/Hydrogenase/MV+. as a reductant, is identical to the resting enzyme. This is the first time that a c-type nitrite reductase has been shown to reduce sulfite. These findings, besides revealing a new function for the nitrite reductase, raise a major question regarding the sulfur metabolism in the sulfate reducing bacteria as well as the cellular localization of the enzymatic activities involved in the dissimilatory reduction of sulfate. The purified nitrite reductase is a heterooligomer, containing two types of subunits of 62 kDa (+/- 5 kDa) and 18.8 kDa (+/- 1 kDa), and forms a complex or aggregate with a molecular mass of approximately 750 kDa. PMID- 8713098 TI - Expression of human lysosomal alpha-mannosidase activity in transfected murine cells and human alpha-mannosidase deficient fibroblasts. AB - We studied the human lysosomal alpha-mannosidase (MANB) by expressing the putative cDNA in mammalian cells, using the eucaryotic expression vector pCDE. The construct pCDE-MANB and pSV2-Neo were cotransfected into human alpha mannosidase deficient fibroblasts and into a murine cell line and selected by culture in the presence of G418. Six G418 resistant 3T3 clones had increased alpha-mannosidase activity 2 to 3 times above the controls. Two clones from transfected human fibroblasts showed a 2 fold increase in enzyme activity. The human MANB cDNA gene was demonstrated in the target cells by Southern blot analysis and the expression of the gene was shown by RT-PCR analysis. This study is the first to successfully express the MANB gene in a human and a murine cell line. The results confirm that the putative MANB cDNA encodes the full length of lysosomal alpha-mannosidase. Molecular characterization of mannosidosis and approaches to gene therapy are now possible using this cDNA. PMID- 8713099 TI - Salt-sensitive hypertension in bradykinin B2 receptor knockout mice. AB - The kallikrein-kinin system regulates water and sodium excretion and thus plays a role in blood pressure (BP) homeostasis. We tested the hypothesis that mice lacking the gene encoding for the bradykinin B2 receptor (B2-KO) have a greater hypertensive response to chronic high Na+ intake (salt sensitivity) compared to controls. We also obtained dose-response curves for different vasoactive substances in both groups. The hypertensive effect of high Na+ intake was almost doubled in B2-KO mice compared to controls. A high-Na+ diet increased heart and kidney weight in B2-KO, but not in controls, suggesting an increased afterload in B2-KO mice. The BP response to bradykinin was completely abolished in B2-KO, but that to acetylcholine was conserved. The hypertensive response to angiotensin II was not exaggerated in B2-KO mice. This study describes a new salt-sensitive animal model and suggests that in mice kinins play a role in preventing salt sensitive hypertension. PMID- 8713100 TI - Tumor necrosis factor alpha receptor p55 is under hormonal (follicle-stimulating hormone) control in testicular Sertoli cells. AB - Identification of TNF alpha receptors and their regulation by the endocrine system were examined in testicular Sertoli cells. By using cross-linking experiments as well as northern blotting analysis, we detected the p55 receptor type (MW: 55 kDa, mRNA: 2.3 kb), but not the p75 TNF alpha receptor type. Furthermore, we report that Sertoli cell TNF alpha receptors are under the control of FSH, but not of other hormones which affect Sertoli cell activity. FSH enhances (ED50: 2 ng/ml) TNF alpha binding to Sertoli cells. This stimulating effect of FSH occurs at the level of p55 TNF alpha receptor, as indicated by the increase in p55 mRNA after FSH treatment. Together, these observations show that the action of TNF alpha (which originates in the testis from germ cells and/or interstitial macrophages) on Sertoli cells remains under the control of the endocrine system as shown by the regulatory effect of FSH on TNF alpha p55 receptors. PMID- 8713101 TI - Functional analysis of the human guanylin gene promoter. AB - Guanylin (GCAP-I, guanylate cyclase activating peptide I) and uroguanylin (GCAP II, guanylate cyclase activating peptide II) are regulatory peptides involved in the regulation of the intestinal chloride / water balance. They share significant structural homology to the E. coli enterotoxin STa, which binds to the particulate guanylyl cyclase C causing diarrhea in mammals. In this study we report the functional analysis of the guanylin / GCAP-I gene promoter region. By means of the luciferase reporter gene assay, we demonstrate a strong promoter activity in T84 cells. Especially the first 160 bp of the 5'-flanking region of the gene seem to be essential for gene induction. Our findings are the basis for further identification of important regulatory elements of the corresponding gene. PMID- 8713102 TI - Phosphatidylserine synthesis in glioma C6 cells is inhibited by Ca2+ depletion from the endoplasmic reticulum: effects of 2,5-di-tert-butylhydroquinone and thimerosal. AB - The effects of 2,5-di-tert-butylhydroquinone (DBHQ) and thimerosal on phosphatidylserine synthesis by the base exchange reaction and on calcium mobilization in intact glioma C6 cells were compared with that of thapsigargin, a selective inhibitor of the endoplasmic reticulum Ca(2+)-ATPase. It has been found that all these agents inhibit phosphatidylserine synthesis by 70%, but their effectiveness are different. The data show that this inhibition is caused by Ca2+ depletion of the endoplasmic reticulum, indicating that phosphatidylserine synthesis requires high concentration of Ca2+ within this structure. On this basis and on literature data, a new model for the localization of the serine base exchange enzyme in the endoplasmic reticulum membrane is proposed. PMID- 8713103 TI - Differential effects of progestins and estrogens on long control regions of human papillomavirus types 16 and 18. AB - We have studied effects of a series of progestins and estrogens on regulation of human papillomavirus (HPV) types 16 and 18 gene expression via long control region (LCR), using a transient expression assay which represented the episomal state of HPVs in precancerous lesions. Results showed that HPV gene expression induced by these steroid analogs were diverse. Gene expression via HPV16-LCR was upregulated 2-3 fold under treatment with 10(-7) M estriol, 17 beta-estradiol, and several progestins including pregnenolone, 17 alpha-hydroxy-progesterone, norethynodrel and cyproterone acetate. However, transcription via HPV18-LCR was not or weakly stimulated by these analogs. Our results can divide these analogs into HPV-enhancing and HPV-nonenhancing groups and accordingly impact the relative risk of individual oral contraceptive ingredient in cervical carcinogenesis. PMID- 8713104 TI - Steady shear and step changes in shear stimulate endothelium via independent mechanisms--superposition of transient and sustained nitric oxide production. AB - We propose that fluid shear presents two distinct stimuli to endothelium-the rate of change of flow and flow itself, to which cells sense and respond via independent mechanochemical transduction pathways. We demonstrate that nitric oxide production occurs by two independent mechanisms; a G protein-dependent transient burst stimulated by rapid changes in flow, and a G protein-independent sustained production under steady or smoothly transitioned flow. The novel use of step, ramp, and impulse flow in this study to stimulate nitric oxide production allows the isolation of these individual production events. Impulse flow activates only the G protein-dependent transient burst, which ramp flow fails to stimulate yielding only the sustained response. Step flow, which contains both a rapid increase and a steady flow component, stimulates both pathways, with the response of the superposition of the transient burst and sustained production. PMID- 8713105 TI - Identification of the 70kD heat shock cognate protein (Hsc70) and alpha-actinin-1 as novel phosphotyrosine-containing proteins in T lymphocytes. AB - T cell antigen receptor (TCR) ligation results in the tyrosine phosphorylation of numerous intracellular protein substrates, and the identification of these substrates has been a major undertaking by several groups. We have used pervanadate treatment to artificially increase cellular phosphotyrosine levels and immobilized anti-phosphotyrosine monoclonal antibodies to partially purify tyrosine phosphorylated proteins in quantities suitable for amino acid sequencing. This strategy was used to identify three phosphotyrosine containing proteins, with relative molecular masses of 105, 81, and 76 kD by amino acid sequencing. Here we report the identification of pp105 as alpha-actinin-1, pp81 as the murine equivalent of the HS1 gene product, and pp76 as Hsc70. This is the first report that alpha-actinin-1 and Hsc70 are targets of activated tyrosine kinases. Furthermore, we show that Hsc70 is tyrosine phosphorylated in response to TCR ligation, which constitutes the first evidence that Hsc70 might be subject to regulation by tyrosine kinase signaling pathways. PMID- 8713106 TI - Biochemical properties of a novel U2AF65 protein isoform generated by alternative RNA splicing. AB - A variety of RNA binding proteins with one or more RNA recognition (RNP-CS) motifs play essential roles in the pre-mRNA splicing process. One such factor, the U2 snRNP auxiliary factor large subunit (U2AF65), contains three RNP-CS motifs each of which is required for high affinity binding to polypyrimidine tracts. Here we report the isolation of a natural cDNA variant of human U2AF65, U2AF65 (S), which is shortened by a 12 nucleotide in frame deletion between RNP CS2 and -CS3 motifs. A portion of the U2AF65 (S) cDNA was reported previously but was not characterized further. We observe that the U2AF65 (S) variant predominates in a variety of tissues and cell lines, and is generated together with the U2AF65 (L) form (2) by alternative 5' splice site selection from a single gene. The corresponding histidine-tagged recombinant proteins bind with similar affinities to model RNA substrates containing strong or weak polypyrimidine tracts. Both U2AF65 (S) and (L) protein isoforms reconstitute splicing activity with similar kinetic profiles in U2AF-depleted (splicing deficient) HeLa nuclear extracts. Finally, the thermal stabilities of the protein isoforms are essentially equivalent. Thus, the presence or absence of the peptide segment, VSPP (residues 345-348), in the linker region between RNP-CS2 and -CS3 does not detract from the intrinsic RNA binding and splicing properties of the U2AF65 protein. The biological implications of alternative splicing for the function and evolution of RNA binding proteins are discussed. PMID- 8713107 TI - Ca2+ induces an increase in cGMP-phosphodiesterase activity in squid retinal photoreceptors. AB - In invertebrate photoreceptors, light elicits the opening of cationic channels to produce a depolarizing receptor potential. One hypothesis is that cGMP is the agent that gates the channels. It has been previously proposed that the light induced rise in intracellular Ca2+ down-regulates phosphodiesterase activity, thereby eliciting an increase in intracellular cGMP concentration. cGMP phosphodiesterase activity from squid photoreceptors was monitored both by a continuous fluorescence assay using 2'-o-(N-methylanthraniloyl)-cGMP and by hydrolysis of [3H]cGMP. The activity of cGMP-phosphodiesterase was found to be increased by Ca2+ in the physiological range of concentrations. These findings suggest that the previously known light-induced increase in Ca2+ in invertebrate photoreceptors cannot directly account for light-elicited down-regulation of cGMP phosphodiesterase. PMID- 8713108 TI - Pseudosubstrate flexibility in chicken smooth muscle myosin light chain kinase. AB - Chicken smooth muscle myosin light chain kinase pseudosubstrate is contained within the C-terminal region of the sequence MLCK(774-807)amide. It forms a helical conformation which interacts with the connecting peptide contained within the N-terminal portion of the sequence. These regions are linked by a well defined turn around Ser-787 which supports a model of pseudosubstrate regulation involving the hinging of the pseudosubstrate or autoregulatory recognition sequence away from the active site of the catalytic core following the binding of calmodulin to the kinase. PMID- 8713109 TI - The reaction between nitric oxide and alpha-tocopherol: a reappraisal. AB - Recently Gorbunov et al. reported that nitric oxide (.NO) can directly oxidize alpha-tocopherol to alpha-tocopheroxyl radical (Gorbunov et al., Biochem. Biophys. Res. Commun., 219, 835-841, 1996). We have reinvestigated this reaction and report that a direct reaction between .NO and alpha-tocopherol does not occur. However, the reaction between .NO and oxygen generates an oxidant which oxidizes alpha-tocopherol to alpha-tocopheryl quinone. Exposure of alpha tocopherol to a low flux of .NO generated from spermine NONOate (100 microM) results in no consumption of alpha-tocopherol under either aerobic or anaerobic conditions. A higher flux of .NO, generated from 1 mM spermine NONOate, oxidizes alpha-tocopherol only under aerobic conditions. Artifactual oxidation of alpha tocopherol can be observed when using commercial .NO that is contaminated with higher oxides of nitrogen, such as dinitrogen trioxide and dinitrogen tetraoxide. PMID- 8713110 TI - Expression of inducible nitric oxide synthase mRNA in rat digestive tissues after endotoxin and its role in intestinal mucosal injury. AB - Nitric oxide (NO) production is increased in the intestine and may contribute to intestinal injury in sepsis. However, the tissue expression of inducible NO synthase (iNOS) mRNA throughout the digestive tract and its relation with the mucosal damage after endotoxin challenge remain unknown. We therefore measured tissue expression of mRNA encoding iNOS by Northern blot analysis and reverse transcription PCR. The iNOS mRNA was detectable at 1 h, peaked at 4 h, and remained faint at 24 h after endotoxin injection in esophagus, duodenum, jejunum, ileum, and colon, but not in the stomach. Pre-treatment with dexamethasone attenuated the rise of iNOS mRNA. Both dexamethasone and NOS inhibitor, L-NAME, ameliorated the endotoxin-induced increase in intestinal mucosal permeability. Our results indicate that there is tissue-specific expression of iNOS mRNA in the digestive tract. The manipulations that decrease NO production may have therapeutic potential in preserving intestinal mucosal integrity in sepsis. PMID- 8713111 TI - Ligand-induced dimerization of the extracellular domain of the TGF-beta receptor type II. AB - Transforming growth factor-beta (TGF-beta) signals by mediating the association of two distinct transmembrane serine/threonine kinase receptors, the type I (T beta RI) and II (T beta RII). Here, we took advantage of recombinant human T beta RII extracellular domain (T beta RII-ED) to analyze TGF-beta/T beta RII complex formation which is the initial event in the construction of a signaling complex. We found that recombinant T beta RII-ED binds TGF-beta 3 more efficiently than TGF-beta 2 and therefore maintains the native T beta RII binding selectivity for the different TGF-beta isoforms. Biochemical analysis showed that free T beta RII ED is expressed as a monomer. Upon ligand binding, both TGF-beta 3 and -beta 2 isoforms induce homodimerization of T beta RII-ED, each TGF-beta subunit being able to bind one T beta RII-ED molecule. These results suggested that ligand dependent receptor dimerization may be an important early step in the TGF-beta signaling complex formation. PMID- 8713112 TI - A novel series of DNA triple helix-binding ligands. AB - We have examined the effect of a series of substituted imidazothioxanthones on the stability of an intermolecular DNA triple helix by DNase I footprinting. We find that several of these compounds promote the formation of a complex between T5C5 and the target site A6G6.C6T6, suggesting that they bind specifically to triplex DNA. The only inactive derivative lacked a protonatable function in the side chain, suggesting that this is an essential feature for triplex stabilization. These compounds, which are amongst the first triplex-binding ligands which possess an uncharged chromophore, are selective for the T.AT rather than the C+.GC triplet. PMID- 8713113 TI - Neokyotorphin and neokyotorphin (1-4): cytolytic activity and comparative levels in rat tissues. AB - The content of biologically active hemoglobin fragment neokyotorphin (TSKYR) as well as that of neokyotorphin fragment (1-4) (TSKY) were determined in extracts of lung, heart, and brain tissue of rats. The content of both peptides as well as the neokyotorphin/neokyotorphin(1-4) ratio differed significantly from each other in these tissues. The respective parameters deviate considerably from those of erythocytes where these peptides are originally formed. Comparative analysis of cytolytic activity of peptides was performed at human erythroid leukaemia (K562) and murine transformed fibroblast (L929) cell lines. TSKY showed reliable cytolytic activity in both cell lines, while neokytorphin was not cytotoxic. The data obtained lead to speculation that endogenous hemoglobin fragments might participate in regulation of tumor growth in vivo. PMID- 8713114 TI - Susceptibility to the development of gastric mucosal damage in OLETF rat not expressing the CCK-A receptor. AB - Otsuka Long-Evans Tokushima Fatty (OLETF) rats are a new model of a congenital defect of the CCK-A receptor gene and should be useful for determining CCK-A receptor function. Since the CCK-A receptor plays an important role in gastrointestinal function, in the present study we examined, using OLETF rats, the hypothesis that a defect of the CCK-A receptor may influence gastric mucosal integrity. The gastric mucosal integrity was evaluated by the severity of gastric mucosal damage induced by ulcerogenic manipulations. The severity of gastric mucosal lesions seen after administration of intracisternal thyrotropin-releasing hormone analogue, subcutaneous indomethacin, intragastric HCl, or intragastric ethanol was evaluated in OLETF and control LETO rats. Administration of thyrotropin-releasing hormone analogue, indomethacin, HCl, or ethanol significantly increased the severity of gastric mucosal lesions in OLETF rats compared with control LETO rats. These results suggest for the first time that gastric mucosal integrity may be impaired in OLETF rats. It is furthermore speculated that individuals that lack of CCK-A receptors are highly susceptible to gastric ulceration. PMID- 8713115 TI - Cyclic AMP stimulates the phosphorylation and activation of p42 and p44 mitogen activated protein kinases in 3T3-F442A preadipocytes. AB - The effects of cyclic AMP on mitogen-activated protein (MAP) kinase activation were investigated in 3T3-F442A preadipocytes. Several agents, which raise intracellular cyclic AMP levels by distinct mechanisms, induced a transient activation of the p42 and p44 isoforms of MAP kinase and of a MAP kinase kinase. Activation of MAP kinase by cyclic AMP was prevented by two distinct inhibitors of cyclic AMP-dependent protein kinase and by PD 098059, a specific inhibitor of the activation of the MAP kinase kinase MEK 1. Therefore, in contrast to most cell types studied, cyclic AMP exerts a positive influence on the MAP kinase pathway in 3T3-F442A preadipocytes at a level upstream of the activation of MAP kinase kinase. PMID- 8713116 TI - Multiple inositol 1,4,5-trisphosphate receptor isoforms are present in platelets. AB - Platelets are activated by an increase in cytosolic Ca(2+), and a portion of this increase is derived from inositol 1,4,5-trisphosphate (InsP3)-mediated Ca(2+) release from internal stores via the InsP3 receptor. There is some uncertainty concerning the localization of the InsP3 receptor within platelets, and experiments were designed to help resolve this question. [3H]InsP3 binding to unphosphorylated and phosphorylated platelet internal membranes revealed both low and high affinity InsP3 binding sites, indicating the presence of more than one isoform of InsP3 receptor within the internal membranes. Phosphorylation did not significantly affect InsP3 binding. In contrast, a single class of high affinity sites was observed in plasma membranes indicating only one type of InsP3 receptor. Western blotting of platelet internal and plasma membranes with antibodies against the three major InsP3 receptor isoforms revealed that the internal membranes contain both type 1 and type 2 InsP3 receptors while the plasma membrane contains only InsP3 receptor type 2. PMID- 8713117 TI - Glutamic acid decarboxylase (GAD 67) gene expression in the pancreas and brain of the nonobese diabetic mouse. AB - A quantitative PCR was developed to measure GAD 67 mRNA levels in pancreas from NOD mice. Two nonautoimmune mouse strains were used as controls. An internal standard, rat GAD 67 mRNA, was prepared to quantify the reaction. In NOD mice GAD 67 expression in the pancreas was in the order of 2 x 10(6) molecules/microgram of total RNA; this expression was five times less in the two control mouse strains, whatever the age. The overexpression of GAD 67 characterized in the NOD mouse could be restricted to the pancreas since similar analysis performed in mouse brain did not reveal a large difference between the three mouse strains studied. PMID- 8713118 TI - Are cytoskeletal components involved in the control of hepatic carnitine palmitoyltransferase I activity? AB - The present work was undertaken to test whether cytoskeletal components are involved in the control of rat-liver carnitine palmitoyltransferase I (CPT-I) activity by cellular effectors. The microtubule stabilizer taxol abolished the changes in CPT-I activity induced by the effectors tested. Taxol also prevented OA-induced shrinkage of hepatocytes as well as the enhanced release of lactate dehydrogenase from digitonin-permeabilized hepatocytes. On the basis of its relative sensitivity to tautomycin and OA, the modulation of CPT-I activity seemed to involve mostly protein phosphatase 1. These data suggest that the short term control of hepatic CPT-I by cellular effectors may involve modulation of interactions between CPT-I and cytoskeletal components. PMID- 8713119 TI - Altered type VI collagen synthesis by keratoconus keratocytes in vitro. AB - Type VI collagen and gelatinase A are synthesized by human keratocytes in vitro. Keratoconus is a corneal disease that has increased gelatinase A activity. Normal keratocyte culture media analyzed by Western blotting with polyclonal antibody to type VI collagen showed a single 140 kDa band that increased in intensity after phorbol ester treatment. The same 140 kDa band was absent in media collected from keratoconus keratocytes cultured with or without phorbol ester. We speculate that in keratoconus keratocyte cultures, type VI collagen may be degraded or modified by the increased gelatinase A activity so as not to be recognized by the type VI antibody. PMID- 8713120 TI - Refolding parameters for the allosteric homodimeric guanylyl cyclase catalytic core from the atrial natriuretic peptide receptor. AB - Protein folding continues to be an important biophysical topic in molecular biology. We report the parameters for successfully refolding the guanylyl cyclase core of the ANP receptor, an allosteric homodimeric enzyme. Urea was a better chaotropic solvent than guanidine HCl, and physiological salt concentrations and pH were needed for optimal recovery of enzymatic activity. Renaturation was more sensitive to alkaline compared to acidic deviations in solvent conditions. The time course of refolding was sigmoidal producing an enzyme with a specific activity of 16,000 pmol cGMP/min/mg using 60 microM concentration of substrate. Additional factors are described in this unusual case of renaturing an allosteric homodimeric enzyme in vitro. PMID- 8713121 TI - Gene disruption of the P-glycoprotein related gene pgpa of Leishmania tarentolae. AB - Transfection of pgpA into Leishmania confers resistance to arsenite and antimonials. By gene targeting mediated by homologous recombination the two alleles of the pgpA gene of a L. tarentolae wild-type cell were disrupted sequentially with the neomycin and hygromycin phosphotransferase genes. This pgpA null mutant showed an increased sensitivity to arsenite and antimonite. In addition, the L. tarentolae pgpA null mutant exhibited a decreased intracellular survival inside murine macrophages. The observed phenotypes were reverted to levels not statistically different than wild-type when an intact pgpA gene was introduced into the null mutant. Disruption of the pgpA chromosomal locus in an arsenite resistant mutant indicated that PgpA is not essential for resistance to oxyanions, although it might be required in the early steps of selection when resistance is being established. PMID- 8713122 TI - Bacillus thuringiensis crystal proteins CRY1Ab and CRY1Fa share a high affinity binding site in Plutella xylostella (L.). AB - The future success of Bacillus thuringiensis based insecticides depends in part on our ability to prevent insects from developing resistance against their insecticidal crystal proteins. Two recent papers indicated cross-resistance between Cry1A proteins and Cry1Fa in two different insect species (Tabashnik et al., 1994, Appl. Environ. Microbiol. 60, 4627-4629; Gould et al., 1995, J. Econ. Entomol. 88, 1545-1559). Brush border membrane vesicles were prepared from Plutella xylostella and used in binding assays with 125I-labeled trypsin activated crystal proteins. Competition experiments showed that Cry1Fa competed with Cry1Ab for a same binding site, though the latter still bound to a different minor binding site with apparently the same affinity. Cry1Ca did not compete for Cry1Ab binding sites nor Cry1Fa for Cry1Ca binding sites. Based on these results, a modification of the receptor shared by Cry1Ab and Cry1Fa might confer multiple resistance to Cry1A and Cry1Fa proteins. PMID- 8713123 TI - The CYP52 multigene family of Candida maltosa encodes functionally diverse n alkane-inducible cytochromes P450. AB - The n-alkane-assimilating yeast Candida maltosa contains several structurally related cytochromes P450 (P450) encoded by the CYP52 multigene family, which are inducible by various long-chain hydrocarbons and fatty acids and which are responsible for the initial hydroxylation steps in the metabolism of these substrates. In the present work, the four major n-alkane-inducible C. maltosa P450 forms; CYP52A3, CYP52A4, CYP52A5, and CYP52A9, were enzymatically characterized, taking advantage of heterologous P450/reductase coexpression in Saccharomyces cerevisiae. Testing various alkanes and fatty acids, distinct preferences of the individual P450 forms concerning substrate class and chain length were detected, thus providing new insight into the functional diversity of the C. maltosa CYP52 family. Moreover, the results obtained emphasize these structurally related enzymes as a powerful tool for future studies on P450 structure-function relationships. PMID- 8713124 TI - High-yield production of Saccharomycopsis fibuligera glucoamylase in Escherichia coli, refolding, and comparison of the nonglycosylated and glycosylated enzyme forms. AB - The truncated GLA1 gene encoding the mature form of glucoamylase from the yeast Saccharomycopsis fibuligera has been over-expressed in Escherichia coli using the IPTG inducible pET system. Over-expression has led to the accumulation of insoluble glucoamylase in inclusion bodies from which an electrophoretically homogeneous active enzyme has been prepared yielding 30 mg per litre medium. This protein represents an N-terminus Met-free, non-glycosylated product which displays the identical specific activity of 45 units/mg and reduced thermal stability when compared to glycosylated enzymes isolated from Saccharomyces cerevisiae carrying the GLA1 gene. These data suggest that S. cerevisiae glycosylation of S. fibuligera glucoamylase does not play a critical role in enzymatic activity but that it does contribute to its thermal stability. PMID- 8713125 TI - Sodium butyrate induces tyrosine phosphorylation and activation of MAP kinase (ERK-1) in human K562 cells. AB - Butyrate is a naturally occurring 4-carbon fatty acid. Biologically, butyrate has been shown to affect the morphology and growth rate of mammalian cells, as well as induce gene expression. Moreover, butyrate has been proven to serve as an anticancer agent, which unlike others (methotrexate and hydroxyurea), is a nontoxic, safe alternative to cancer treatment. It also induces erythroid differentiation in K562 cells. However, its mechanism of action has yet to be determined. In this study we investigated the effects of sodium butyrate (NaB) on tyrosine phosphorylation in K562 erythroleukemic cells. We demonstrate that NaB induces both dose and time-dependent tyrosine phosphorylation of several proteins, the effects of which were blocked by the tyrosine kinase inhibitor genistein. Furthermore, NaB induces tyrosine phosphorylation and rapid activation of MAP kinase (ERK-1). These findings provide the first evidence that the signal transduction mechanism of NaB involves rapid tyrosine phosphorylation and activation of MAP kinase. PMID- 8713126 TI - Induction of dystrophin-associated proteins together with nicotinic acetylcholine receptors by denervation in the absence of dystrophin in skeletal muscles of mdx mice. AB - We purified the nicotinic acetylcholine receptor from digitonin-solubilized rabbit skeletal muscle by affinity chromatography and detected many proteins linked to AChR, including dystrophin, adhalin, beta-dystroglycan, utrophin, rapsyn, and actin. To determine whether or not AChR links to dystrophin associated proteins (DAPs) without dystrophin, we studied the effects of denervation on AChR and DAPs in the skeletal muscle of a mdx mouse. Following surgical denervation, the levels of adhalin and beta-dystroglycan dramatically increased at the extrajunctional sarcolemma with AChR, suggesting that their association is independent of dystrophin. Furthermore, the diffuse extrajunctional appearance of adhalin, beta-dystroglycan, and AChR was observed after pharmacological denervation through the subcutaneous administration of succinylcholine. Since the depletion of DAPs and the subsequent disruption of sarcolemmal linkage are believed to be a primary cause of muscle cell necrosis in dystrophinopathies, pharmacological denervation may have some beneficial effect on these diseases. PMID- 8713127 TI - Shear stress-induced release of PGE2 and PGI2 by vascular smooth muscle cells. AB - This study addresses the direct effect of fluid flow shear stress on production of the vascular mediators, PGE2 and PGI2 by vascular smooth muscle cells (SMC). Results indicate that shear stress increases PGE2 and PGI2 release in SMC. The production patterns, however, differ between PGE2 and PGI2. For PGE2, the rate of production is moderate for the first three hours after the onset of shear stress, then dramatically increases between the fourth and fifth hours, returning to basal levels in the sixth hour. On the other hand, the rate for PGI2 production is maximal right after the onset of shear and remains elevated for the first three hours. The rate then plateaus and remains at a moderate level during the next three hours. The results also indicate that SMC production of PGI2 is more sensitive to shear stress than PGE2 production since a level of 0.5 dynes/cm2 produces a maximal PGI2 release whereas 1 dyne/cm2 produces only 1/4 the response seen at 20 dynes/cm2 for PGE2. The physiological implications of fluid shear stress regulation of SMC are discussed. PMID- 8713128 TI - A novel strain, B/Gifu/2/73, differs from other influenza B viruses in the receptor binding specificities toward sialo-sugar chain linkage. AB - We reported in a previous paper (1. Xu et al, J. Biochem., 115, 202-207, 1994) that every human influenza B virus isolate determined, including B/Lee/40, B/Setagaya/3/56, B/Tokyo/7/66, B/Kagoshima/1/68, B/Kanagawa/3/76, B/Ibaraki/2/85, B/Yamagata/16/88, and B/Bangkok/163/90, restrictively recognizes lacto-series gangliosides containing Neu5Ac alpha 2-6Gal linkage. We found in this study a novel strain, B/Gifu/2/73, that showed remarkable receptor binding specificities to both Neu5Ac alpha 2-3 Gal and Neu5Ac alpha 2-6Gal linkages of lacto-series gangliosides. The amino acid sequence of the hemagglutinin (HA) in strain B/Gifu/2/73 was compared with that of B/Lee/40, B/Ibaraki/2/85, B/Yamagata/16/88, and B/Bangkok/163/90. Substitution of His116, Ala121, Arg141, Pro211, His253, and His271 in the HA1 subunit of strain B/Gifu/2/73 for Asn116, Thr121, Gly141, Gln211, Gln253, and Gln271 in the HA1 of the four strains above was found, indicating that these amino acid changes in the HA1 of B/Gifu/2/73 may play an important role in recognition of Neu5Ac alpha 2-3Gal and Neu5Ac alpha 2-6Gal linkages in the receptor sialo-sugar chains. PMID- 8713129 TI - Accumulation of 5-aminolevulinic acid-induced protoporphyrin IX in normal and neoplastic human endometrial epithelial cells. AB - The aim of this study was to evaluate 5-Aminolevulinic acid (ALA)-induced fluorescence of normal and neoplastic endometrial epithelial cells for diagnosis and photodynamic treatment. Fluorescence of ALA-induced PpIX in vitro was measured by flow cytometry in two different human endometrial adenocarcinoma cell lines and in normal cells cultivated from fresh endometrial tissue of three premenopausal patients. The cells were analysed after incubation with different concentrations of ALA during 3, 6, or 24 hours. Both tumor cell lines showed a statistically significant higher fluorescence of PpIX than normal epithelial cells after incubation with 1 mg ALA per ml medium during 24 hours. The well differentiated cancer cells produced significantly more PpIX than the poorly differentiated cancer cells. Relative PpIX intensity of the two cancer cell lines correlated with cell proliferation rate as measured by the doubling times of the cells. Higher accumulation of Pp IX in neoplastic endometrium compared to normal endometrial epithelial cells may provide targeted biopsies and selective photodynamic destruction of neoplastic micro-lesions. PMID- 8713130 TI - E-selectin binding promotes neutrophil activation in vivo in E-selectin transgenic mice. AB - E-selectin is a membrane protein expressed by endothelial cells activated by cytokines released during the inflammatory process; it plays an important role in neutrophil emigration into inflamed tissues. To further explore in vivo the function of E-selectin, we have generated transgenic mouse line expressing E selectin under the control of a chicken beta-actin promoter. In these mice, the number of blood neutrophils was reduced, without any other obvious phenotype or tissue damage. These neutrophils, however, displayed two significant changes: first, an alteration in the levels of expression of two membrane receptors involved in neutrophil adhesion to endothelial cells, namely a marked increased in the Mac-1 antigen (CD11b/CD18) and a decrease in the Mel-14 antigen (L selectin); second, an increased oxidative activity when compared to blood neutrophils of non-transgenic mice, as shown by their capacity to oxidize 2',7' dichlorofluorescein (DCFH) into a fluorescent compound. These observations indicate that the binding of E-selection with neutrophils bearing its ligands promotes neutrophil activation in vivo. PMID- 8713131 TI - Structure of catechol 2,3-dioxygenase gene encoded in chromosomal DNA of Pseudomonas putida KF715. AB - A catechol 2,3-dioxygenase gene in chromosomal DNA of P. putida KF715 was cloned and its nucleotide sequence analyzed. The enzyme gene was composed of 924 base pairs with ATG initiation codon and TGA termination codon, which can encode a polypeptide of molecular weight 35 kDa containing 307 amino acids. A promoter like sequence and a ribosome-binding sequence were identified upstream the enzyme gene. A deduced amino acid sequence of the catechol 2,3-dioxygenase exhibited 94% identity with that of corresponding enzyme in TOL plasmid and 25% identity with that of 2,3-dihydroxybiphenyl 1,2-dioxygenase from the same strain. Furthermore, sequence comparison of the catechol 2,3-dioxygenase with other extradiol-type dioxygenases has led to identify evolutionally conserved amino acid residues whose possible catalytic roles are proposed. PMID- 8713132 TI - Differential effects of retinoic acid (RA) and N-(4-hydroxyphenyl) retinamide (4 HPR) on cell growth, induction of differentiation, and changes in p34cdc2, Bcl-2, and actin expression in the human promyelocytic HL-60 leukemic cells. AB - Incubation of the HL-60 cells with 3 microM of RA and 4-HPR resulted in suppression of cell growth and decrease in cell viability. A significant percentage of the RA-treated cells also displayed differentiation towards neutrophils, as assayed by changes in nitroblue tetrazolium reduction (NBT) and alpha-naphthyl-acetate esterase (ANAE) activities, whereas the 4-HPR treated cells remained essentially undifferentiated. Flow cytometric analysis showed 4 HPR to cause partial cell arrest in the G2/M phase after a 3-day treatment and an additional G1 phase arrest after a 7-day treatment. With RA-treated cells, a reduction in the percentage of cells in the G1 phase was observed after 7 days of treatment. In 4-HPR-treated cells an extra peak, characteristic of cells undergoing apoptosis, was found in the cell cycle phase distribution analysis. Determination of specific protein expression changes by Western blot analysis showed that the p34cdc2 was down-regulated by both chemicals. Furthermore, RA induced bcl-2 but prevented the processing of actin, whereas 4-HPR had little effect on bcl-2 but increased the specific processing of actin. These results suggest that RA promotes neutrophil differentiation and the establishment of a semi apoptosis-resistant state, possibly through the overexpression of the bcl-2 gene. By contrast, 4-HPR may trigger apoptosis by inducing overall cyto architectural changes and specific DNA fragmentation subsequent to increased turnover of the protein actin. PMID- 8713133 TI - Mosaic type of the nontoxic-nonhemaggulutinin component gene in Clostridium botulinum type A strain isolated from infant botulism in Japan. AB - The gene encoding the nontoxic-nonhemaggulutinin (NTNH) component was amplified by the PCR technique using two primer sets and the DNA template from Clostridium botulinum type A strain 7I03-H isolated from infant botulism in Japan. The nucleotide sequence revealed that the NTNH gene was composed of 1,193 amino acids with a molecular weight of 130868.08. Furthermore, the N-terminal half side and C terminal half side of the NTNH component were similar to the NTNH component of type C and type A, respectively. These results indicate that the NTNH component gene codes the mosaic NTNH component composed of type A and type C. The hemaggulutinin gene, aha, and ORF-22 gene, orf-22a, were undetectable in the region upstream of the NTNH component gene, ant. Therefore, orf-22a is not thought to play a key role in the expression of botulinum type A progenitor toxin gene. PMID- 8713134 TI - Effect of cystathionine and cystathionine metabolites on the phosphorylation of tyrosine residues in human neutrophils. AB - The effect of cystathionine and cystathionine metabolites found in the urine of patients with cystathioninuria on the phosphorylation of tyrosine residues was studied with human peripheral blood polymorphonuclear leukocytes. Among the cystathionine metabolites, cystathionine ketimine markedly increased phosphorylation of a 45 kDa protein with time and the phosphorylation depended on the concentration of cystathionine ketimine, while cystathionine and the reduced form of cystathionine ketimine (cyclothionine) did not increase the phosphorylation of the 45 kDa protein. The phosphorylation of the 45 kDa protein induced by cystathionine ketimine was inhibited by genistein and herbimycin A, inhibitors of tyrosine kinase, but was not inhibited by 1-(5 isoquinolinesulfonyl)-2-methylpiperazine and staurosporine, inhibitors of protein kinase C. PMID- 8713135 TI - A novel gene iba1 in the major histocompatibility complex class III region encoding an EF hand protein expressed in a monocytic lineage. AB - We successfully isolated a rat cDNA clone encoding a novel EF hand protein with a molecular weight of about 17 kDa and designated this gene iba1 (ionized calcium binding adapter molecule 1). The genomic copy of the iba1 gene was located within a segment of the major histocompatibility complex class III region between the Bat2 and TNF alpha genes. The iba1 gene was shown to be highly expressed in testis and spleen, but weakly expressed in brain, lung, and kidney. Among brain cells, the iba1 gene was specifically expressed in microglia. A screening of hemopoietic cell lines showed that the Iba1 protein was clearly expressed in monoblastic cell lines but only very weakly expressed in myeloid cell lines. Iba1 protein is therefore suggested to act as an adapter molecule, mediating calcium signals that may function in a monocytic lineage including microglia. PMID- 8713136 TI - Bisphosphonates are incorporated into adenine nucleotides by human aminoacyl-tRNA synthetase enzymes. AB - Bisphosphonates are synthetic pyrophosphate analogues and are therapeutic inhibitors of bone resorption, although their exact mechanisms of action are unclear. Some bisphosphonates can be metabolised into non-hydrolysable ATP analogues by Dictyostelium discoideum amoebae, in a back-reaction catalysed by several Class II aminoacyl-tRNA synthetases. We have found that the same enzymes in cell-free extracts of several human cell lines are also capable of metabolising in vitro the same bisphosphonates that are metabolised by Dictyostelium. These results indicate that human cells, following drug internalisation, should be capable of metabolising certain bisphosphonates. The toxic effects of these bisphosphonates towards bone-resorbing osteoclasts may therefore be due to accumulation of non-hydrolysable ATP analogues or inhibition of aminoacyl-tRNA synthetase enzymes. PMID- 8713138 TI - Ethically important distinctions among managed care organizations. PMID- 8713137 TI - zhx-1: a novel mouse homeodomain protein containing two zinc-fingers and five homeodomains. AB - A cDNA encoding the complete reading frame of a novel homeodomain-containing protein, named zhx-1, has been cloned from a mouse bone marrow stromal cell line. The ORF encodes a protein of 873 amino acids, in which two C2-H2 zinc-fingers and five homeodomains have been identified. These features classify zhx-1 as a member of the ZF class of homeodomain transcription factors, zhx-1 mRNA is widely expressed in adult mouse, although it is preferentially expressed in brain. At least, two homologous mRNAs are present in humans suggesting that zhx-1 is the first member of a novel subclass of homeodomain factors. PMID- 8713139 TI - The legal subtext of the managed care environment: a practitioner's perspective. PMID- 8713140 TI - Business vs. medical ethics: conflicting standards for managed care. PMID- 8713141 TI - Moral justice and legal justice in managed care: the ascent of contributive justice. PMID- 8713142 TI - Managed care as regulation: functional ethics for a regulated environment. PMID- 8713143 TI - Authority in ethics consultation. PMID- 8713144 TI - Ethics consultation: persistent brain death and religion: must a person believe in death to die? PMID- 8713145 TI - Breast implants: the debate goes on. PMID- 8713146 TI - Sailing with the winds of change. AB - The theme for the 1994 Operating Room Nurse Week in the United States was, "We cannot change the wind, but we can adjust the sails." For the sailor, the wind is a powerful force. It must be constantly monitored and the sails adjusted to keep the vessel on course. Occasionally, if conditions are severe, the course may have to be altered. PMID- 8713147 TI - It's time to change visiting policies in critical care units. AB - For years, hospitals have been highly structured organizations--a state often attributed to their early associations with the Church and military. Nurses have come to accept hospitals' many policies and procedures as part of their everyday lives. Little wonder, then, that it has taken considerable time and energy to question and dismantle policies that were accepted as military orders or gospel from a greater authority. PMID- 8713148 TI - Floating: sink or swim. AB - Budget restrictions, fewer human resources and fluctuating patient populations have combined to increase the practice of floating general duty nurses. Floating occurs when a nurse from an overstaffed unit, based on patient load, is required to work in another unit that is understaffed. Floating is not a new practice, but it has always been an emotionally charged issue. Today, with more float pool nurses seeking the security of a permanent position in a home unit, the issue has increased in intensity. PMID- 8713149 TI - [Consent for organ transplantation]. AB - When patients are trying to decide whether to consent to organ transplantation or refuse treatment, respect for their legal rights is the minimum ethical consideration. Their decision must be a fully informed one. This article focuses on the differences between nurses' and physicians' perceptions of the patient's decision to undergo transplant surgery. The research is based on a survey of eight transplant teams in the province of Quebec. Eight physicians and eight nurses answered a questionnaire and participated in semi-structured interviews. The data obtained was then compared and interpreted using two action theories developed by Kohlberg and Gilligan. This approach enabled the authors to clarify various concepts relating to fairness, as interpreted by the two professional perspectives. The results underline that nurses view things differently than physicians. Their professional approach is based mainly on psychosocial criteria to ensure an adequate response to the expressed or stated needs of the patient. The fact that they do not make decisions regarding the cure sets them apart from physicians, who believe they are offering the best solution to a patient's problem, while respecting the patient's right to informed consent. PMID- 8713150 TI - The law, nurses and coffee breaks. AB - During nurses' shifts, whether they are seven or 12 hours long, breaks are scheduled. The number and time of these breaks varies among institutions but is usually reflected in collective agreements or employment contracts. These breaks are important so nurses can, rest but they need to be scheduled and taken with care. The three cases reviewed below outline the responsibilities of both the charge nurse and staff nurse when scheduling and taking breaks. PMID- 8713151 TI - Shiftwork strategies. AB - Nurses who learn to manage their sleep and diet while rotating shifts may feel a lot better. A pilot project aimed at improving shiftworkers' sleeping patterns and diet resulted in significant improvements in sleep, general health and psychological well-being. The five-month project, involving 10 oncology nurses in Edmonton, Alberta, included a workshop on sleep and diet, and strategies to address individual shiftwork-related problems. PMID- 8713152 TI - Community health for Rwandan refugees. AB - Health education and disease prevention programs are essential elements in every health care system. We normally envision community health programs in urban and rural settings across Canada. However, health education and disease prevention have also become a fundamental part of the health care system for refugee communities around the world. PMID- 8713153 TI - The gift of compassion. AB - Christmas Day in a hospital is always different from Christmas Day anywhere else. For one thing, there is a sense of isolation; family and friends are elsewhere and every patient who can be discharged, even temporarily, leaves for the day. PMID- 8713154 TI - Are science and caring compatible? AB - Scientific progress has led us to believe we are masters of our destiny and in control of our world. But, science and technology can destroy our humanity if they serve no higher power than their own advancement. We can be overwhelmed by a surplus of information that clouds our thoughts and hinders us as we try to find true meaning and purpose in our lives. PMID- 8713155 TI - Making tough decisions. AB - It is amazing that so much money is spent on health care with little understanding and measurement of its effect on people's health status. But tightened health care budgets and the explosion of new and often expensive health care technologies have forced governments, providers and taxpayers to examine how best to maximize health dollars. This has led to greater reliance on economic evaluations to help make difficult choices. PMID- 8713156 TI - Arborization of the inferior laryngeal nerve and internal nerve on the posterior surface of the larynx. AB - The morphological patterns of the inferior laryngeal nerve and internal laryngeal nerve display complex arborizations. This paper attempts to identify and clarify these patterns. Dissections were performed on 105 adult Japanese cadavers, and observations were made on 201 sides. Results showed that the communications between the inferior laryngeal nerve (ILN) and internal laryngeal nerve (ITLN) could be classified into two types and three subtypes. Also, the ITLN displayed three characteristic patterns at the arytenoid cartilage. These communications produce complex arborizations of the ILN as it enters the larynx. This may explain the variety of potential clinical symptoms observed after thyroid surgery or neck dissections. PMID- 8713157 TI - Variations in thickness of articular cartilage in the human sacroiliac joint. AB - Differences in articular cartilage thickness in the sacroiliac joint were investigated in different regions of the sacral and the iliac articular surfaces in the embalmed cadavers of five males and six females. The mean thickness of the sacral articular cartilage was greater than that of the iliac articular cartilage (P < 0.001) and the sacral articular cartilage of the female was thicker than that of the male (P < 0.02). Differences between thicknesses of the iliac articular cartilage in the male and female and in different regions of the sacral and iliac articular cartilages were found to be not significant. PMID- 8713158 TI - Extensor tendons of the fingers: arrangement and variations--II. AB - One hundred eighty-one dissected hands were examined to study the pattern of extensor tendons on the dorsum of the hand. Extensor digitorum often had multiple tendons for the middle and ring fingers. Its contribution to the little finger was usually by a bifurcating tendon common with that of the ring finger. The index finger always received a single tendon. Intertendinous connections between the various tendons of the extensor digitorum were variable but were most frequent between ring and middle fingers. Extensor indicis had one tendon in most of the specimens and it was always on the ulnar side of the extensor digitorum tendon. This remained true even when there were multiple tendons. Extensor digiti minimi had two tendons in most cases. It was always linked to extensor digitorum either by receiving one or part of its tendon or by an intertendinous connection. Two accessory muscles were seen, one was extensor indicis brevis replacing the proper muscle. The other, the extensor medii brevis, was distributed to the middle finger. PMID- 8713159 TI - Topography of the perforating veins on the medial side of the leg (Cockett's veins). AB - A macroscopic anatomical investigation was carried out on 67 embalmed dissecting room legs in order to determine the position, course, and origin of so-called Cockett's perforating veins. This information was considered to be of particular importance because of the frequency with which varicoses in this region are treated with sclerosing agents and the serious adverse effects that can follow a misplaced injection. In view of the almost universal credence given to the accuracy of Cockett's original description (Cockett FP, 1955, Br. J. Surg. 43:260 278), and the general acceptance of the constancy of "Cockett's veins," it was felt to be of importance to see to what extent we could confirm this description. It was found that the origins of these vessels and the places at which they penetrate the deep fascia are not constant, and that--in our series--only 7.3% of the veins belonging to these groups occupy the positions that are repeatedly cited in the literature as the most common. The clinical importance of any discrepancy between expectation and reality can hardly be exaggerated. PMID- 8713160 TI - The buccal pad of fat: a review. AB - Within the cheek, wedged between masseter and buccinator, is a biconvex pad of fatty tissue, the corpus adiposum buccae, or buccal fat pad (of Bichat). It contributes significantly to the prominence of the cheek of the newborn infant and is sometimes encountered in surgical procedures in the region of the ramus of the mandible or the maxillary tuberosity. This paper reviews the history of the study of the buccal pad of fat, its anatomical location, blood supply, and comparative anatomy. We have also reviewed the pathology of the buccal pad of fat, including traumatic herniation. The fat pad is of interest surgically as it can be used as a free or pedicled graft to close maxillary defects after excision of tumors. PMID- 8713161 TI - A graphic, three-dimensional teaching model to demonstrate the topography of the human heart. AB - A three-dimensional graphic model of the human heart and an accompanying description are presented as a teaching aid. Each of the four chambers is represented by a cube of equal size. Drawings of the model from various angles give views of each of the surfaces and show the interatrial, interventricular, and atrioventricular grooves, and superimposed coronary arteries and cardiac veins. PMID- 8713162 TI - Rotation of dissecting groups among different cadavers provides students of anatomy with increased opportunity to observe human variations. AB - One way to increase medical students' awareness of anatomical variability is the dissection of different cadavers throughout laboratory coursework. This report covers such a procedure successfully instituted in a human gross anatomy course. PMID- 8713163 TI - What future for dissection in courses of human topographical anatomy in universities in the UK? AB - In the United Kingdom most medical schools have traditionally included dissection as a major component of their anatomy courses. There is a dearth of substantive literature which can be used to demonstrate the educational values of dissection, although many authors have expressed opinions. Current scrutiny of the role of dissection has been prompted by many influences acting on medical schools in the UK, including changes in the medical curriculum promoted by the General Medical Council, changes in the mechanisms of funding for universities and their courses, and the introduction of research assessments and appraisal of teaching. It is probable that in the UK dissection will continue to decline but that its reduced role will be prompted by the interactions of many complicated processes, only some of which will be educationally driven. PMID- 8713164 TI - The terminology of early development: history, concepts, and current usage. AB - The meaning of some terms used in describing the development of embryos is discussed in the light of their historical origin and current usage. Attention is focused on the terminology associated with the early development of animals, particularly that concerned with gastrulation and germ layer theory. It is suggested that terms linked with outdated concepts are not always appropriate for, and unless defined with care may limit, our appreciation of the significance of subsequent observation and experiment. Recommendations are made concerning the use of the terms mesoblast, mesoderm, and mesenchyme. PMID- 8713165 TI - The official nomenclature of the superficial veins of the lower limb: a case for revision. AB - Attention is called to the discrepancy which exists between the official terminology of the Nomina Anatomica (PNA) as it relates to the superficial venous system of the lower limb, and anatomical reality. Suggestions are also put forward as to how the PNA could be altered to accord better with the systematic classification and topography of these vessels, and more usefully serve the needs of clinicians. PMID- 8713166 TI - An English translation of Alzheimer's 1907 paper, "Uber eine eigenartige Erkankung der Hirnrinde". PMID- 8713168 TI - Anatomy of the valve of the coronary (Thebesian valve) PMID- 8713167 TI - Anomalous flexor digitorum superficialis presenting as a painful palmar mass: a case report. AB - This case study presents an anomalous flexor digitorum superficialis indicis. The patient presented with a painful palmar mass. Physical exam and exploratory surgery were used to describe this anomaly. The anomalous muscle was intrinsic to the hand. PMID- 8713170 TI - Crossbridge dynamics under various inotropic states in cardiac muscle: evaluation by perturbation analyses. AB - As is evident in this review, significant advances have been made in the analysis of crossbridge cycling in cardiac muscle. It becomes apparent that the myocardial crossbridge cycling rate is altered by various factors such as: 1) changes in the intracellular constituents (Ca2+, MgATP, MgADP, Pi, and H+), 2) difference in the type of myosin isozyme, and 3) probably, troponin I, C-protein, and MLC2 phosphorylation. The physiological consequences of an altered crossbridge cycling rate may give rise to the changes in the contraction profile during twitch, tension cost (the ratio of ATPase activity to tension), and the thermal economy (the ratio of heat liberated to tension) of cardiac muscle. These findings and implications should be kept in mind when interpreting cardiac performance under different inotropic states. PMID- 8713171 TI - Modulation of phagocytosis by P2-purinergic receptors in mouse peritoneal macrophages. AB - The effects of adenosine 5'-triphosphate (ATP) and other nucleotides on phagocytosis of peritoneal macrophages were examined by means of flow cytometry (FCM). ATP and uridine 5'-triphosphate (UTP) enhanced phagocytosis in a dose dependent manner. Control phagocytosis was partially suppressed in Ca(2+)-free solution. Phagocytosis was suppressed by ATP in Ca(2+)-free solution but not by UTP. Agonist potency order revealed that ATP enhanced phagocytosis through P2u receptors in normal external solution (NES) and inhibited it through P2z receptors in Ca(2+)-free solution. The present study showed that ATP modulates the phagocytosis of macrophages, and that two types of receptors were identified by pharmacological experiments. PMID- 8713172 TI - Photometric determination of phenomenological correlation between osmotic behavior and hemolysis of red blood cells. AB - The osmotic behavior of red blood cells from a human and from several other mammalian species was studied by photometric measurements. When red blood cells were suspended in sodium chloride solutions with various osmotic concentrations, the optical density at 620 nm was reciprocally related to the relative volume of the red cells. Thus, we evaluated the osmotic volume changes in the red cells from optical density measurements. The Boyle-van't Hoff relation was applicable to the osmotic behavior of red cells which responded as a complete osmometer in hypertonic and slightly hypotonic (lower than about 240 mOsm) solutions. Also, we examined the rheological correlation between osmotic volume changes and hemolysis. Osmotic hemolysis occurred corresponding to breakdown of the Boyle van't Hoff relation in hypotonic solutions. The critical osmotic concentration for the breakdown of the Boyle-van't Hoff relation was that for osmotic hemolysis. In Na2SO4 solutions, although the critical osmotic concentration shifted towards a smaller value, the critical volume for the breakdown of the Boyle-van't Hoff relation and for osmotic hemolysis was maintained at a constant value, indicating that the onset of osmotic hemolysis depends exclusively upon the critical volume. In the samples from a human, the critical volume for the onset of hemolysis was estimated to be 1.25 +/- 0.05 in the ratio to the normal volume in iso-osmotic solution. From these obtained results, it is suggested that the red cell behaves in hypotonic solutions as a viscoelastic body of the type represented by the Voigt model, and the viscoelastic breakdown of the membrane results in osmotic hemolysis in hypotonic solution. PMID- 8713173 TI - Regional hemodynamic changes and vasopressin release induced by intracisternal injection of L-proline in the conscious rat. AB - Electromagnetic flow probes were used to measure regional blood flow responses to central injection of L-proline in conscious rats. Ten min after intracisternal injection of 10 mumol of L-proline, arterial pressure increased from 113 +/- 1.6 (mean +/- SEM, n = 23) to 142 +/- 2.9 mmHg, while the heart rate decreased from 383 +/- 9.6 (n = 23) to 313 +/- 5.4 beats/min, vascular resistance increased an average of 239% in the superior mesenteric artery (n = 8) and 72% in the renal artery (n = 8), without change in the terminal aorta (-6%, n = 7). However, 10 mumol of D-proline induced neither blood pressure increase nor bradycardia 10 min after its injection. After sino-aortic denervation, the heart rate was not decreased by L-proline injection. Ganglionic blockade with chlorisondamine evoked significantly greater pressor action by L-proline injection than that without treatment, but intravenous injection of the vasopressin antagonist, (d(CH2)5(1), O-Me-Tyr2, Arg8)-vasopressin (10 micrograms/kg), promptly dilated the superior mesenteric vascular bed and lowered the arterial pressure. In addition, pretreatment with the vasopressin antagonist alone almost abolished the pressor and vasoconstrictor action induced by L-proline injection. These results indicate that intracisternal injection of L-proline specifically elicited an increase in arterial pressure mainly through marked vasoconstriction of the superior mesenteric artery bed, and that bradycardia was elicited via the baroreflex. Thus, vasopressin release in the blood-stream was involved in these hemodynamic actions. PMID- 8713174 TI - Hindquarter vasodilation after intracisternal injection of D-arginine in the conscious rat. AB - The cardiovascular effects of the centrally injected D-arginine were investigated in the conscious rat chronically instrumented with cisternal, arterial, and venous cannulae, as well as an electromagnetic flow probe around the superior mesenteric artery, renal artery, or terminal aorta (hindquarter). Intracisternal injection of D-arginine (10 mumol) increased arterial blood pressure and hindquarter flow lasting for 60 min or more. Previous injection of the beta adrenoceptor blocker, propranolol (2 mg/kg, I.V.), markedly attenuated the hindquarter vasodilation caused by D-arginine; this response suggests the release of adrenaline. Peripheral resistance, calculated as arterial pressure divided by regional flow, increased in both the superior mesenteric artery (60%) and the renal artery (45%) 5 min after intracisternal injection of D-arginine. In the hindquarter, however, peripheral resistance decreased by 35% and arterial pressure increased by 25%. The pressor effect was significantly attenuated by producing ganglionic blockade with chlorisondamine (5 mg/kg, I.V.). The total peripheral blood flow increased from 12.9 to 15.9 ml/min/100 g body weight. This response indicates that the pressor effect of D-arginine is due to an increase of cardiac output rather than of peripheral resistance. Centrally administered D arginine appears to activate the sympatho-adrenal system, especially the release of adrenaline. PMID- 8713175 TI - The central effect of L-cysteine on cardiovascular system of the conscious rat. AB - The hemodynamic effects of intracisternal injection of the nonessential amino acid L-cysteine were studied in conscious chronically instrumented rats. Injections of L-cysteine (0.05-0.2 M in artificial cerebrospinal fluid, 10 microliters) into the cisterna magna dose-relatedly elicited an increase in arterial pressure but a decrease in superior mesenteric blood flow as measured by an electromagnetic flow probe. Injections of the excitatory amino acid transmitter L-glutamate at comparable doses caused much the same pressor and vasoconstrictor effects as did L-cysteine. Prior I.V. injection of vasopressin V1 receptor antagonist, (d(CH2)5(1), O-Me-Tyr2, Arg8)-vasopressin (10 micrograms/kg), markedly attenuated the effects of L-glutamate but not of L cysteine. Ganglionic blockade with chlorisondamine (5.0 mg/kg) failed to attenuate the effects of either amino acid, whereas an additional intravenous injection of vasopressin antagonist, completely abolished the effects. These results indicate that the circulatory effects of L-cysteine are probably due to autonomic nervous activation combined with vasopressin release, unlike those of L glutamate which acts mainly through vasopressin release. L-Cysteine may contribute to central cardiovascular control, since it induces the marked circulatory effects comparable to or greater than those of L-glutamate. PMID- 8713176 TI - Proximal tubular cell alkalinization induced by Na(+)-glucose cotransport. AB - To study the ionic mechanisms of crosstalk between luminal and peritubular membranes induced by Na(+)-glucose cotransport in the proximal tubular cells, we applied the ion-selective microelectrode techniques and measured intracellular ion activities of Na+ (Nai) and H+ (pHi), as well as peritubular membrane potential (EM) in doubly-perfused bullfrog kidney. In control conditions with peritubular HCO3- (15 mM), luminal perfusion with 10 mM D-glucose induced a 12 mV depolarization of peritubular membrane (glucose-induced depolarization) with a transient increase in Nai by 3 mM and an elevation in pHi by 0.1. In the absence of peritubular HCO3- (HEPES-Ringer at constant pH), the luminal glucose produced a sustained increase in Nai and suppression of pHi elevation without affecting glucose-induced depolarization. Peritubular perfusion with DIDS (10(-4) M) abolished the change in pHi induced by luminal glucose. Furthermore, the luminal glucose increased the transport number for K+ (gK/gm) in the peritubular membrane by 20%. These results suggest that 1) glucose-induced depolarization suppresses peritubular HCO3- exit, followed by an elevation in pHi, 2) the luminal glucose increases K+ conductance of the peritubular membrane, probably by the elevation in pHi, and 3) this increased K+ conductance repolarizes the peritubular membrane, which can support the driving force for Na(+)-glucose entry across the luminal membrane. Thus, we conclude that the crosstalk between peritubular and luminal membranes by Na(+)-glucose cotransport is mainly modulated by changes in cell pH and membrane potential in the proximal tubule. PMID- 8713177 TI - Effects of electrical stimulation of cervical sympathetic trunks on microcirculation in the facial nerve. AB - This study evaluates the circulatory effects of electrical stimulation of the cervical sympathetic trunks on blood flow in the common carotid artery and facial nerve tissue in dogs. Marked increases in arterial pressure and heart rate were observed due to electrical stimulation of the cervical sympathetic trunks, while blood flow volume in the common carotid artery and in the facial nerve tissue decreased markedly. It was assumed that microcirculation of the facial nerve is definitely impaired by electrical stimulation of the cervical sympathetic trunks, and the tonicity of the sympathetic nervous system appears to be a major factor in changes in the microcirculation of the facial nerve. It is well known that impaired circulation in the nutrient vessels of the facial nerve has an important effect on the pathogenesis of facial palsy. The hypertonicity of the sympathetic nervous system is closely involved in the onset of facial palsy. PMID- 8713178 TI - Chemical modification by maleimide of toxic and nontoxic ouabain actions on neuromuscular transmission in the frog. AB - A dual action of ouabain on the Mg(2+)-blocked frog neuromuscular transmission was studied in two different experimental procedures by measuring the mean quantal content (m) of endplate potentials evoked during repetitive nerve stimulation. The nontoxic action of low-dose (1 microM) ouabain was observed as a counterclockwise pivoting of the linear stimulation frequency versus the log m relation, termed frequency augmentation-potentiation (FAP), whereas the toxic effect of higher doses (> or = 10 microM) of ouabain was recorded as a slow exponential increase in m. Since neural Na+,K(+)-ATPase consists of two or three isoforms, the nontoxic action of low-dose ouabain was anticipated to be a consequence of the selective inhibition of ouabain-sensitive alpha(+) (alpha 2 and alpha 3 or neural type) Na+,K(+)-ATPase activity. Such a possibility was assessed by applying epsilon-maleimidocaproic acid (MCA), a membrane-impermeant sulfhydryl reagent. MCA 15 microM was found in the present study to suppress the nontoxic effect of ouabain without altering the toxic effect. The results strongly suggest that the toxic and nontoxic actions of ouabain are the result of actions on different entities. The ouabain-sensitive neural type Na+,K(+)-ATPase, which is abundant in neural tissues, seems to play an important role in modulation of transmitter release observable as FAP. PMID- 8713179 TI - The effects of aging on the rat bladder and its innervation. AB - 1) Measurements of the cystometrogram, of the responsiveness of bladder muscle to pelvic nerve efferent stimulation and of the sensitivity of the pelvic nerve afferents to pressure and volume during distensions have been made in the bladders of young adult (2-3 months) and aged (26-29 months) rats, anesthetized with mixtures of urethane and chloralose. 2) The pressure-volume relationship differed in young adult and aged rats. The bladders of the aged rats held up to nearly six times the volume of the young animals, and these volumes were accommodated at lower pressures in the aged animals. The pressure at which micturition contractions appeared was similar in young adult and aged animals. 3) The passive pressure associated with each of a series of distending volumes was recorded when a pelvic nerve was cut unilaterally. The distal cut end of this cut pelvic nerve was stimulated for 10 s at 20 Hz, using square wave pulses of 10 V and 1.0 ms. The active pressure-volume relationship was constructed from this data. Both the active and the passive relationships were shifted to the right in the aged animals, and it was evident that aging was associated with a reduction in the maximal pressure generated during pelvic nerve stimulation. Also the change in intravesical pressure induced by bladder contraction was less in aged animals. 4) The most sensitive mechanoreceptor afferents appear to have pressure and volume thresholds that do not change significantly during aging. While the distension-sensitive afferents in the pelvic nerve appear to have a similar sensitivity to intravesical pressure in young adult and aged rats, they were less able to monitor volume in the aged animals. The stimulus response relationship for volume was often less steep in the aged animals. 5) In this study, aging was shown to be associated with a large increase in bladder volume and a reduced sensitivity of pelvic nerve afferents to volume, and a reduced ability to raise bladder pressure during contraction of bladder smooth muscle. The changes in bladder function associated with aging are discussed. PMID- 8713180 TI - Sinusoidal and exponential decays of postextrasystolic transient alternans in excised blood-perfused canine hearts. AB - We have recently reported that the postextrasystolic contractile potentiation decays in alternans after a compensatory pause in canine left ventricles even under normal coronary and contractile conditions. The transient alternans appears to consist primarily of a small-magnitude exponential decay and a large-magnitude sinusoidal decay. We, therefore, hypothesized that the contractility (y) of the postextrasystolic alternans beats (beat number x) could be expressed as y = a x exp[-(x-1)/b] + c x exp[-(x-1)/d] x sin[pi(x-0.5)] + yo, where a and c are the normalized magnitudes (relative to the preceding regular beat) of the two exponential terms in the first postextrasystolic beat, b and d are their time constants, and yo is the normalized magnitude of the post-alternans regular beat (approximately 1). The first exponential term represents the monotonic decay. The sine term multiplied by the second exponential term represents the alternating decay. Mathematical curve-fitting indicated: 1) the above equation very closely fitted the alternans data with a squared correlation coefficient of 0.9996 on average, 2) c was 7 times on average greater than a, indicating dominance of the sine component, 3) b and d were 2.5 and 1.0 beats on average, indicating a faster decay of the sine component, and 4) this b was comparable to the time constant of the exponential decay of the postextrasystolic potentiation after no compensatory pause. This study suggests that myocardium has a mechanism to switch the postextrasystolic potentiation between the exponential and alternans decays depending on the first postextrasystolic interval. PMID- 8713181 TI - Responses mediated by excitatory and inhibitory amino acid receptors in solitary spiking cells from normal newt retina. AB - Whole-cell currents activated by L-glutamate, kainate (KA), alpha-amino-3-hydroxy 5-methyl-4-isoxazole-propionic acid (AMPA), N-methyl-D-aspartate (NMDA), gamma aminobutyric acid (GABA), and glycine were recorded from spiking cells enzymatically dissociated from adult newt retina. Most spiking cells responded to both AMPA and KA. KA tended to be a more potent agonist for them than AMPA. The reversal potential of both AMPA and KA responses was about -7 mV; this value was similar to that (about -5 mV) of glutamate response. AMPA- and KA-induced currents may be carried by monovalent cations, such as Na+, because their reversal potentials were sensitive to external Na+. About half of the spiking cells examined responded to mixtures of NMDA and glycine. Extracellular Mg2+ blocked completely the response to NMDA plus glycine in spiking cells held at negative membrane potentials, but not at positive membrane potentials. Almost all spiking cells responded to the inhibitory amino acids GABA and glycine. Dose dependent desensitization was observed in both GABA and glycine responses. Currents activated by GABA and glycine were carried by Cl-. Bicuculline and strychnine strongly suppressed the responses to GABA and glycine, respectively, suggesting the existence of GABAA receptors and conventional glycine receptors in the spiking cells. PMID- 8713182 TI - Development of responses to excitatory and inhibitory amino acids in spiking cells during retinal regeneration in the adult newt. AB - Responsiveness to amino acids in solitary spiking cells dissociated from regenerating newt retinae at different stages were studied by whole-cell patch clamp methods in comparison with that in the normal retina. Cells from regenerating retinae of 1-2 cell thickness ("early" stage) lacked the receptors for kainate (KA), alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid (AMPA), N-methyl-D-aspartate (NMDA), gamma-aminobutyric acid (GABA), and glycine. AMPA/KA, NMDA, GABAA, and glycine receptors, each of which were electrophysiologically and pharmacologically similar to those in the normal retina, appeared in spiking cells before regenerating retina had segregated into two synaptic layers. During subsequent regeneration stage, the time course of appearance of amino acid receptors seems to be different between excitatory and inhibitory ones. Cells which responded to GABA and glycine gradually increased in number to normal levels, and the current amplitude induced by these amino acids also increased monotonically. The number of cells responding to the excitatory amino acids appeared to be maximal at a period of synaptic segregation. During subsequent regeneration, however, cells responded to AMPA and NMDA decreased in number and returned to the number of cells in the normal retina, while cells which responded to KA were maintained at the same number as those in the normal retina. The amplitudes of the current induced by excitatory amino acids also became maximal at a period of synaptic segregation. During subsequent regeneration, AMPA- and KA-induced currents declined to the normal level, while NMDA-induced currents were maintained. PMID- 8713183 TI - Decline in maximal oxygen uptake on work performance in rats during the developmental phase. AB - Development-related changes in maximal oxygen uptake (VO2max) and work performance were examined in young sedentary rats 4-11 weeks after birth during exercise. The running speed to elicit VO2max increased from 4 to 8 weeks of age, whereas the exercise VO2max declined progressively. Therefore, the work performance during the developmental phase, when rapid growth occurs, seems to be little related to the decline in the relative VO2 max. PMID- 8713184 TI - Benzamil-induced conductive state of insulin-stimulated, amiloride-blockable cation channel in fetal lung epithelium. AB - Insulin activated a 28 pS amiloride-blockable nonselective cation (NSC) channel in rat fetal distal lung epithelium. Benzamil, an analogue of amiloride, decreased the open probability (Po) of the insulin-unstimulated channel from 0.06 +/- 0.02 to 0.0013 +/- 0.0006 (mean +/- SEM, n = 5-7; 100 microM benzamil application to extracellular surface), but increased the Po of the insulin stimulated channel from 0.10 +/- 0.02 to 0.69 +/- 0.02 (mean +/- SEM, n = 5-7; 100 microM benzamil application to extracellular surface). The effects of benzamil could be observed in either case that it was applied to the extracellular or cytosolic surface. Unlike benzamil, amiloride decreased the Po of both insulin-unstimulated and -stimulated channels. These observations suggest that benzamil acts as a blocker on the insulin-unstimulated NSC channel but as an opener on the insulin-stimulated NSC channel. PMID- 8713185 TI - Increased muscle sympathetic nerve activity during delayed auditory feedback in humans. AB - We examined the effect of mental stress on muscle sympathetic nerve activity (MSNA) by microneurographically measuring the MSNA of the tibial nerve while subjects read aloud. A delayed auditory feedback task (DAF) was employed with a delay interval of 200 ms and these performances were compared to those in the absence of DAF in five healthy men (28 +/- 2 years). The mean blood pressure was significantly elevated during reading both in the presence of DAF and in its absence, and the pressor response was more exaggerated in the presence of DAF than its absence. MSNA and plasma norepinephrine levels were significantly increased during reading in the presence of DAF but not in its absence, whereas heart rate was mildly increased at the same extent during reading in both the presence and absence of DAF. The results suggest that mental stress produces elevation in blood pressure and mediates the activation of sympathetic nerve activity. PMID- 8713186 TI - Does recurrent inhibition exist in the paraventricular magnocellular neurosecretory cells in the male rat? AB - Extracellular recordings were performed from the paraventricular (PVN) neurosecretory cells in the anesthetized male rats. After sub- and suprathreshold stimulations to the neural stalk, two of 38 neurons showed inhibitory phases and eleven of the remaining 36 neurons showed inhibitory responses after suprathreshold stimulations. These results indicate that the recurrent inhibitory system exists in the PVN. PMID- 8713187 TI - The public health workforce education and training study. PMID- 8713188 TI - Medical education and health for all: dream, mirage or nemesis? PMID- 8713189 TI - See no evil, fix no evil. PMID- 8713190 TI - Diabetes in aborigines and Torres Strait Islanders: effects, causes and strategies. PMID- 8713191 TI - Prevalence of cigarette smoking among Australian secondary school students in 1993. AB - The prevalence of cigarette smoking in Australian secondary school students in 1993 was estimated from a survey of 22 696 12- to 17-year-old students from 332 secondary schools (and feeder schools) in all states and the Northern Territory. Self-administered questionnaires were answered anonymously by groups of up to 20 students selected randomly from school rolls, a method that replicated previous surveys in 1984, 1987 and 1990. Current smoking (smoking at least one cigarette in the week preceding the survey) at 12 years of age was 8 per cent in boys and 7 per cent in girls, but in those 17 years of age the prevalence was much higher (28 per cent of boys, 31 per cent of girls). Age was also associated with the mean number of cigarettes smoked per week by current smokers (8.6 in boys and 7.0 in girls at age 12 and 43.8 in boys and 32.0 in girls at age 17). After controlling for sex, age, school type and state of residence, the percentage of 12- to 15-year-olds who were current smokers rose from 15.7 per cent to 17.5 per cent between 1990 and 1993, an effect that was more pronounced in boys. On the other hand, the mean number of cigarettes smoked by 12- to 15-year-old current smokers dropped from 23 per week to 19.5, and the reduced consumption was greatest in boys. Furthermore, there was no increase in the proportion of students who smoked on three or more days per week, which suggests the increase was limited to occasional, casual or social smoking. PMID- 8713192 TI - The cost-effectiveness of compulsory bicycle helmets in New Zealand. AB - This paper examines the cost-effectiveness for primary school children (age 5-12 years), secondary school children (13-18 years) and adults (over 18 years) of the legislation enacted on 1 January 1994 requiring road-cyclists in New Zealand to wear helmets. The cost to cyclists not in possession of a helmet before they became compulsory of either obtaining one or quitting cycling was compared with the number of deaths and hospitalisations expected to be prevented over the average life of a helmet. Corresponding to Victorian and United States estimates of the efficacy of cycle helmets at preventing serious head injuries, the cost per life saved was $88 379 to $113 744 for primary school children, $694 013 to $817 874 for secondary school children, and $890 041 to $1 014 850 for adults (New Zealand dollars = approximately 0.95 Australian dollars). The cost per hospitalisation avoided was $3304 to $4252, $17 207 to $20 278, and $49 143 to $56 035 respectively. These estimates are extremely sensitive to the estimated efficacy of helmets at protecting cyclists. Mainly anecdotal evidence for New Zealand suggests that they are not to be very effective at preventing serious head injuries; future research into the change in injury patterns as a result of the helmet regulation would be valuable. Nonetheless, the ranking of the abovementioned estimates does not contradict the policy in some parts of the world requiring helmets for children and/or teenagers, but not adults. PMID- 8713193 TI - Declining blood lead levels in Victorian children. AB - To investigate the distribution of blood lead levels in a sample of Victorian children, and to compare current levels with those from a similar survey in 1979, blood was tested for lead in 252 children (123 under five years) attending Royal Children's Hospital as outpatients and having venepuncture blood samples for medical reasons. Blood lead levels were determined by graphite furnace atomic absorption spectrophotometer. The mean blood lead level was 0.26 mu mol/L (5.4 mu g/dL). In the under-five age group, the mean was 0.28 mu mol/L (5.7 mu g/dL). Only 1.6 per cent of this group exceeded the National Health and Medical Research Council action level of 0.72 mu mol/L (15 mu g/dL). Levels in this age group have declined significantly since 1979, when the mean was 0.54 mu mol/L (11.1 mu g/dL) and 12.9 per cent exceeded 0.72 mu mol/L (15 mu g/dL). Average blood lead levels have halved since 1979, with likely contributing factors being reduced exposure from lead in diet, reduced access to lead in paint and reduced lead in ambient air. Children with elevated levels had identifiable risk factors such as pica or exposure to lead-based paint, suggesting the need for ongoing pubic health action to prevent exposure in these groups. PMID- 8713194 TI - Increasing attendance at immunisation clinics: lessons from a trial program that failed. AB - Previous studies suggested that the use of a baby-enrolment and reminder system for early childhood immunisation increased public immunisation clinic attendance. It was decided to run a trial to assess the effects of introducing a baby enrolment program on attendances at local government immunisation clinics. Enrolment leaflets were distributed to each mother of a new child in six areas of greater Sydney while the mother was in the postnatal ward of the local hospital. Clinic attendance figures were monitored, interviews were conducted with mothers as well as professionals involved in early childhood health services, and the conduct of clinics was observed. The trial failed to increase attendance at public clinics. Mothers' reasons for choice of service, particularly their desire for what they believed to be a more personalised service, and the attitude of the professionals (particularly community nurses) with whom they came in contact were considered to be more influential in determining their use (or lack of use) of public immunisation services. Baby enrolment may be effective in increasing attendance at public immunisation clinics only where there is willing cooperation of all stakeholders in supporting public immunisation services. PMID- 8713195 TI - A population-based survey of immunisation coverage in two-year-old children. AB - A cross-sectional, population-based, cluster-sample survey of 187 children was conducted in the Newcastle area to assess the proportion of two-year-old children who were fully immunised, to ascertain whether administration of these vaccines was age-appropriate and to look for factors predicting incomplete immunisation. Parents or guardians were interviewed at their homes and the immunisation status of the children was verified either by the parent-held record or by the immunisation-provider-held record. Levels of full immunisation were 77 per cent at the time of interview and 72 per cent at the second birthday if Haemophilus influenzae type b (Hib) vaccine was excluded. If it was included, the full immunisation level was 51 per cent. Coverage was highest for oral polio vaccine and lowest for Hib vaccine. Twenty-nine per cent of all immunisations were given early, 44 per cent were given on time and 20 per cent were given late. Doses of vaccines due at older ages were more likely to be given late or not at all. Factors predicting incomplete immunisation were: the principal caregiver being aged under 25 years, being born outside Australia, having post-secondary qualifications, being female and having more than one child in the household. Immunisation coverage levels were not high enough to protect against outbreaks of pertussis and measles and cases of Haemophilus influenzae type b. Immunisation providers should aim to increase coverage to protect the population against all vaccine preventable diseases, and aiming at high-risk groups could more effectively do this. PMID- 8713196 TI - Injury morbidity in Victoria among adults 25 to 64 years of age: implications for prevention. AB - The aim of this retrospective study was to complete the baseline descriptive epidemiology of hospital-treated injury for all ages in Victoria, by focusing on the age group, 25 to 64 years, to identify major categories of injury and the potential for prevention. We analysed injury databases for hospital admissions and emergency department presentations for major variables influencing injury frequency and rates. We found that nearly 40 per cent (158 537) of all hospitalisations for injury occurred in the study age range. The major causes of injury were health-care related causes (32 per cent), falls (15 per cent), transport (14 per cent), and intentional causes (10 per cent). The leading reason for emergency department presentation (excluding admission) was unintentional cutting or piercing injury. The home was the major location of injury. Vehicles, man-made and natural surfaces, knives, and floors and flooring materials were among the leading five factors potentially associated with both admissions and presentations. Injury prevention in this age range will be largely addressed by the injury-prevention strategy being implemented in Victoria. However, specific additional issues were identified, including falls from ladders and scaffolds, pedestrian and female passenger injury, and injury associated with power tools, chain saws, and knives. PMID- 8713197 TI - A community-based cervical screening program in a remote Aboriginal community in the Northern Territory. AB - We established a culturally appropriate, community-based women's health service in Yuendumu, Northern Territory, to improve women's health and to remedy the low rate of cervical screening. During the 16 months of the program 419 cervical smears were taken, increasing coverage of the women eligible from 51 to 78 per cent. Acceptance of the program was excellent, with only 2 per cent of the women approached refusing to have a smear. Over 70 per cent of the Pap smears were done by the nursing staff in the clinic; quality control was good, with 9 per cent of smears reported as having no endocervical cells. Sixty-four per cent of screened women had normal smears and 0.9 per cent showed evidence of cervical intraepithelial neoplasia (CIN). Three women were referred for treatment of CIN, two for CIN I and one for CIN III. The program illustrates how a combination of community involvement, staff stability, teamwork, and cross cultural understanding can achieve a comprehensive and successful cervical screening service in a remote Aboriginal community. PMID- 8713198 TI - High mortality from renal disease and infection in Aboriginal central Australians with diabetes. AB - Few studies have examined the consequences of the high prevalence of diabetes in Aboriginal communities. We aimed to determine the rates and causes of mortality in all Aboriginal central Australians with diagnosed diabetes, identified by a previous study (n =374). Cohort members were followed from 1 January 1984, or the date of diagnosis (to 31 December 1986), to 31 December 1991 or death. Death certificates, medical notes and autopsy reports were examined for cause of death. There were 130 deaths in 2280.7 person-years of follow-up. Standardised mortality ratios for Aboriginal people with diabetes, compared to the Northern Territory Aboriginal population, were 209 (95 per cent confidence interval (CI) 158 to 273) for men and 169 (CI 129 to 218) for women. The difference in ratios for men and women was not statistically significant when adjusted for age (P = 0.2). The eight-year survival rates for men and women diagnosed between 1984 and 1986 were 55.8 per cent (CI 32.6 to 73.7) for men and 80.3 per cent (CI 64.8 to 89.5) for women. Renal disease was the direct cause of death in 22.3 per cent. Infection accounted for 20.8 per cent of deaths and ischaemic heart disease for 13.8 per cent. Forty-four per cent of death certificates made no mention of diabetes. Diabetes confers an additional risk of death on a population whose mortality is already markedly worse than that of other Australians. Unlike Western diabetic populations, infections and renal disease were more common causes of death than macrovascular disease. Diabetes amplifies the effect of the community prevalence of infection and renal disease. PMID- 8713199 TI - Aborigines and tuberculosis: why they are at risk. AB - Aborigines have higher rates of Mycobacterium tuberculosis than the rest of the community. There are insufficient contemporary data to assess how much risk tuberculosis poses to the Aboriginal community. Tuberculosis is of particular concern because of its interaction with human immuno-deficiency virus (HIV). We aimed to ascertain the available data about tuberculosis in Australian Aborigines: to determine morbidity and mortality of tuberculosis in Australian Aborigines, to ascertain the extent of known risk factors for tuberculosis in Australian Aborigines and to consider the public health implications of our findings. Sparse evidence suggests that Aborigines have higher rates of infection and of clinical tuberculosis than non-Aboriginal Australians, along with a high prevalence of known risk factors for tuberculosis. However, there is a paucity of data about specific risk factors and tuberculosis in Aborigines. In addition, Aborigines have a high prevalence of risk factors for HIV infection. The existence of concurrent risk factors for tuberculosis and HIV, in a population that already has a high rate of infection with tuberculosis is cause for grave concern. Tuberculosis control is centred on correct and rapid diagnosis and appropriate treatment, as well as efficient contact tracing. These are the most important strategies for control of tuberculosis among Aborigines, and are especially important when there is concurrence of other risk factors. Appropriate preventive therapy for infected people should also be considered. PMID- 8713200 TI - Infection control and human immunodeficiency virus: perceptions of risk among nurses and hospital domestic workers. AB - In December 1993 the first case of patient-to-patient transmission of human immunodeficiency virus (HIV), at a doctor's surgery in New South Wales, was documented. In an environment of heightened anxiety about HIV transmission and the adequacy of infection-control measures taken by health providers, it is important to explore perceptions of occupational risk of exposure to infection among hospital workers, reasons why hospital domestic workers sometimes depart from standard procedure in infection control, and how they regard the patients who have infectious diseases. In this study, at an infectious diseases hospital where there is an acute awareness of such issues, nurses had accurate knowledge about control of infection, including HIV, but had limited trust of that knowledge. They gave rationales for why they sometimes departed from infection control procedures. They had low levels of fear of homosexuals and of acquired immune deficiency syndrome. The hospital domestic workers had lower levels of accurate knowledge about infection control, including HIV, and less trust of that knowledge and of protection by health provider from occupational exposure to infection. They had low levels of fear of homosexuals and HIV. Both groups sought regular, small-group, interactive education programs on infection control and HIV to allow them to discuss their concerns. Participatory education of workers should include eliciting concerns of participants, and should discuss concerns regarding administrators' and educators' interests in their safety and wellbeing. PMID- 8713201 TI - Medical and health stories on the Syndey Morning Herald's front page. AB - Front-page coverage of medical and health stories in the Sydney Morning Herald over the one-year period, April 1992 to March 1993, was analysed. Features of the front-page stories, such as major topics, geographic location, visual imagery, use of news actors and news sources, the representation of medicine and health, the use of language in headlines, and dominant and recurring discourses were examined. Although front-page coverage was often contradictory and paradoxical, allowing space for alternative views on the value or otherwise of medical treatment and preventive health measures, it was predominantly conservative, giving greater voice to elite groups than less powerful groups, such as advocacy, activist and community groups, and to men rather than women. It tended to individualise illness rather than place it in its broader socioeconomic and political contexts. PMID- 8713202 TI - Conducting regional health surveys using a computer-assisted telephone interviewing method. AB - After the creation of thirteen health regions within Queensland in 1991, the need arose for an information base at the regional level to assist regions with their role in planning, monitoring and evaluating health services. A series of regional health surveys was conducted in 1993 to provide this information, using a computer-assisted telephone interviewing method. Over 10 400 interviews were conducted throughout the state. This is the first time a computer-assisted method has been used on a large scale to collect health-related information in Australia. Interviews used list-directed or random-digit dialing, depending on the rate of unlisted numbers in a region. Response rates were not significantly different for the two methods, although the number of contactable numbers attempted and the noncontact rates were significantly higher for random-digit regions. The last-birthday method was used to select the adult for interview in each household. The method resulted in a bias toward female respondents. PMID- 8713203 TI - Prevalence and frequency of health service use: associations with occupational prestige and educational attainment. AB - The accessibility of health care services has been suggested to be one factor with the potential to ameliorate the health effects of socioeconomic disadvantage. From a randomly selected sample of households in the Lower Hunter Valley region, 2623 adults were surveyed in 1987-88 to identify their reported use of medical, allied and alternative health services during the previous four weeks. There was a higher prevalence of use of the 'usual' general practitioner and medical services among educationally disadvantaged respondents only. No significant differences were evident between educational or between occupational groups in the prevalence of use of either alternative services or health services generally. Fewer occupationally disadvantaged respondents reported using allied health services. There was no difference in the number of health services used. Disadvantaged respondents were more likely to use medical services exclusively. Only educationally disadvantaged service users reported using any health, medical or general practitioner services more frequently than expected. In contrast, only occupationally disadvantaged service users reported using allied health services (and allied health services other than dentists) more frequently. The lack of consistent differentials in use across health services in favour of disadvantaged respondents suggests that a number of health care services may not be responding to the greater need for health care among disadvantaged members of the community. PMID- 8713204 TI - Malignant mesothelioma in Pilbara Aborigines. AB - Malignant mesothelioma occurred in a female Aborigine after environmental exposure to asbestos. All known cases of the disease in Aborigines in Western Australia were reviewed; all occurred in Pilbara residents. Most were exposed while involved in the transport of asbestos from the Wittenoom crocidolite operation. Based on recent estimates of the size of the Aboriginal population in the Pilbara region, their incidence of this disease (250 per million for ages 15 and over) is one of the highest population-based rates recorded. PMID- 8713205 TI - Australian dietary targets in 1995: their feasibility and pertinence to dietary goals for 2000. AB - To ascertain whether the diet of young students in health-related courses conformed to Australian dietary targets for 1995 and to identify nutritional needs in view of dietary targets for 2000, 246 undergraduate students analysed their weighed diet for three or five days. Eight of ten male participants were meeting the dietary goals for 1995 for fibre and fat intake and six of ten were meeting the goals for 2000. More female participants were meeting the goals for fat, but less than half of the female were meeting the 1995 goals for fibre. Some 84 per cent of females, but only 29 percent of males, were meeting the 1995 dietary goals for sodium. Few participants derived 10 per cent or less of their energy from saturated fat or 10 per cent of their energy from polyunsaturated fatty acids. Substantial numbers of participants failed to meet the recommended daily intakes of zinc. calcium, magnesium and iron (females). Education of specific subgroups and modifications to food production and processing are proposed. PMID- 8713206 TI - Medical education and health for all. PMID- 8713207 TI - Public health in the graduate medical curriculum at Flinders University. PMID- 8713208 TI - Medical education at the University of Sydney. PMID- 8713209 TI - Medical education at the University of Queensland. PMID- 8713210 TI - State differences in Aboriginal health. PMID- 8713211 TI - Choice of denominator in studies of cigarette purchases by minors. PMID- 8713212 TI - Plain English for the journal. PMID- 8713213 TI - Oral health care in Australia. PMID- 8713214 TI - Should the association of physicians of India adopt family planning? PMID- 8713216 TI - Vasculitic neuropathy: profile of twenty patients. AB - Twenty patients with vasculitic neuropathy were analyzed. Sixteen of the twenty presented with classic mononeuritis multiplex but four had distal, symmetrical, sensorimotor polyneuropathy. Though vasculitic neuropathy is classically associated with Collagen vascular syndromes like, polyarteritis nodosa, rheumatoid arthritis and systemic lupus erythematosis, only 13/20 of our patients had definitive Collagen vascular disease. A large proportion (7/20) had vasculitic neuropathy as the only clinical feature. PMID- 8713215 TI - Leptospirosis in Madras--a clinical and serological study. AB - Leptospirosis was confirmed by Microscopic Agglutination Test (MAT) and/or ELISA in 57 patients admitted to the Government General Hospital, Madras, India, during November and December of 1990 and 1991 with symptomatology suggestive of the disease. Fifty (88%) of the 57 cases were males; the mean age of all the cases was 39.6 years (range 17-72). The main clinical features were: fever 100% jaundice 84%, Myalgia 82%, acute renal failure 72% and conjunctival suffusion 58%. Non-azotemic jaundice occurred in 19% of cases. Renal failure was non oliguric in 24% of cases. 3.5% of patients died. 23 patients underwent peritoneal and/or hemodialysis. ELISA IgM titres ranged from 1:80 to 1:10240 (geometric mean tire 911). MAT titres > or = 1:1600 and > or = 1:800 occurred in 39 of 54 and 51 of 54 cases respectively. Autumnalis was the serogroup most commonly recorded serologically, and Leptospira interrogans serovar autumnalis was isolated from one patient. This study shows that leptospirosis is a significant health problem in Madras, though normally grossly underestimated due to the absence of routine laboratory diagnostic facilities for the disease. Gross under-reporting is also likely in other high rainfall third world areas. PMID- 8713217 TI - Study of serum prealbumin and serum alpha fetoprotein in cases of fulminant hepatic failure. AB - We studied serum prealbumin (SPA) and serum alpha fetoprotein (AFP) to assess liver cell injury and prognosis in patients with fulminant hepatic failure (FHF). We studied 21 patients of FHF of viral etiology, 10 acute viral hepatitis (AVH) and 10 healthy controls. Initial (on the day of admission) AFP levels were significantly elevated in FHF group (30.28 +/- 63.58 ng/ml, p < 0.01 compared to AVH and control group in whom it was undetectable. Serum AFP correlated well with deranged Liver Functions (LFT). In the Survivors (n = 4) of FHF, serial estimations (on Day 5 and Day 10 of admission) revealed declining AFP Levels, correlating with clinical recovery. SPA on admission was significantly reduced in FHF group (15.10 +/- 9 mg/dl p < 0.05) compared to AVH (37.0 +/- 9.34 mg/dl) and control group (40.25 +/- 5.92 mg/dl). Low SPA also correlated with deranged LFT. Serial estimations in the survivors (Day 5, Day 10) revealed rising SPA which correlated with clinical recovery. PMID- 8713218 TI - A study of autonomic neuropathy in diabetes mellitus in relation to its metabolic control. AB - 50 Patients of diabetes mellitus (both IDDM and NIDDM) were selected with typical symptoms, signs and positive bed side tests of autonomic neuropathy. All the patients were followed for three months during which strict metabolic control was achieved by routine treatment with oral hypoglycaemic agents and/or insulin, simply by change in their previous treatment dosages and better attention to diet and physical activity. 22% patients showed significant improvement in symptoms of autonomic neuropathy. 42% showed partial improvement and 36% patients did not show any improvement. Improvement in objective test score was significant in 18%, partial in 46% and insignificant in 36%. Improvement in neuropathy did not correlate with HbA1C levels. 36% patients did not show any subjective or objective improvement in autonomic neuropathy inspite of good glycaemic control as indicated by normal HbA1C levels in them. PMID- 8713219 TI - Efficacy of zinc therapy in prevention of crisis in sickle cell anemia: a double blind, randomized controlled clinical trial. AB - 145 patients were recruited in the trial while 130 completed it. Patients were randomized to receive zinc sulphate capsules. 220 mgm three times a day or identical placebo. Major outcome variable was 'Sickle cell crisis'. After a follow up of 1.5 years, the mean number of episodes of crisis was 2.46 +/- 1.04 in the intervention group and 5.29 +/- 2.58 in the control group (p < 0.025; 95% CI for difference between groups: 1.98, 3.42). Mean duration of hospital stay was 4.3 +/- 2.2 days in the intervention group and 3.9 +/- 1.6 days in the control group. The difference was not significant (p > 0.05). There was a significant reduction of the mean number of infective episodes and associated morbidity in patients with sickle cell anaemia. PMID- 8713220 TI - Evaluating efficiency of bone marrow harvest and its manipulation--role of CD 34 positivity. AB - We evaluated harvested marrow cells for total nucleated cells (27.49 x 10(9)), absolute 'lymphocyte' count (6.29 x 10(9)) and CD 34 positive cells (3.57 x 10(9)). The same parameters were studied after in vitro manipulation to remove RBCs and plasma. Reinfused WBCs contained 12.87 x 10(9) nucleated cells, 4.25 x 10(9) absolute 'lymphocyutes' and 3.34 x 10(9) CD 34 positive cells. The corresponding figures for loss during in vitro manipulation (tubing, RBCs and plasma) are 14.62 (53.18%), 2.04 (32.43%) and 0.23 x 10(9) (6.44%) cells respectively. Therefore CD 34 positivity may be a better indicator of total yield, loss during manipulation and reinfusion of hemopoietic progenitor cells in bone marrow transplantation. PMID- 8713221 TI - Circulating immunoglobulin levels in pulmonary tuberculosis. PMID- 8713222 TI - Tolerability of enalapril in mild to moderate hypertension. PMID- 8713223 TI - Neurocysticercosis--an update. PMID- 8713224 TI - Myasthenia gravis. PMID- 8713225 TI - Magnetic resonance imaging is essential for cerebral venous thrombosis. PMID- 8713226 TI - Peripheral neuropathy due to small vessel vasculitis. PMID- 8713227 TI - Survival in thrombotic thrombocytopenic purpura. PMID- 8713228 TI - Rheumatoid vasculitis in India: a report of ten patients. PMID- 8713229 TI - Fibrous dysplasia of bone. PMID- 8713230 TI - Pulmonary manifestation in ankylosing spondylitis. PMID- 8713231 TI - Atypical case of cerebral venous thrombosis diagnosed by MRI. PMID- 8713232 TI - Therapeutic arterial embolization for control of severe hepatic haemorrhage. PMID- 8713233 TI - The memorable patient who changed our practice. PMID- 8713234 TI - Hypereosinophilic syndrome. PMID- 8713235 TI - Urinary symptoms in rabies. PMID- 8713236 TI - Microalbuminuria in non-insulin dependent diabetes and essential hypertension--a marker of severe disease. PMID- 8713237 TI - Withdrawal of antiepileptic drugs in epileptics. PMID- 8713238 TI - Laser hazards and prevention. PMID- 8713239 TI - Cytogenetic study of primary amenorrhoea. AB - A study of chromosomal pattern was done in 60 cases of primary amenorrhoea of different age groups to determine the incidences of chromosomal abnormalities in them and to detect those cases of classical 45, X Turner's syndrome and Turner mosaics that do not bear the Turner stigmata. Buccal smears were examined for sex chromatin followed by karyotype using leucocyte culture method. Majority (63.3%) of cases were found chromosomally incompetent of which the major abnormality was 45, X/46, XX mosaicism (33.3%) followed by 45, X Turner's syndrome (26.6%). But only 43.7% of these Turner's syndrome had classical Turner stigmata. Two cases of complete testicular feminisation syndrome with male genotype (46,XY) and inguinal testis were also detected. PMID- 8713240 TI - Role of antibiotics in clean wounds. AB - Two hundred twenty cases with clean surgical wounds were treated without administering any antibiotics. Majority of them (134) were hydroceles and hernias. The study included some major operation also. The infection rate was 3.6% in the present series, which is comparable with the study where antibiotics were used. PMID- 8713241 TI - Distribution of age-specific rotavirus antibody in human: a hospital-based study in Calcutta. AB - Sera obtained from 332 non-diarrhoeic individuals belonging to different age groups were tested by enzyme-linked immunosorbent assay method for detection of rotavirus antibody. All the sera were found to contain rotavirus antibody in varying titres, the highest being recorded in neonates which declined to the lowest by the age of 6 months with a rapid rise during the next 5 years. This study indicates the acquisition of rotavirus antibody in different age groups. PMID- 8713242 TI - Psychosocial aspect of planned induction of labour. AB - One hundred patients of planned induction and 25 cases of spontaneous delivery were interviewed on the various aspects of labour care in the postpartum period. Majority were satisfied with overall care and 65% were of the opinion of opting for induction next time. PMID- 8713243 TI - Transferable drug resistance in Salmonella typhi strains isolated from an outbreak at Calcutta in the recent past. AB - Transferable drug resistance, phage type, biotype and minimum inhibitory concentration to different drugs were studied in 50 Salmonella typhi strains isolated during recent outbreak of enteric fever at Calcutta. The antibiotic sensitivity test showed that 44% strains were sensitive to multiple drugs including chloramphenicol. The minimum inhibitory concentrations of the drugs tested were well above the normal level. The transferable drug resistance test suggested that all the resistant strains were carrying transferable multidrug resistant gene containing ampicillin, chloramphenicol, streptomycin, tetracycline resistance factor. From phage typing it was observed that 30% strains belonging to phage type 51, and among them 80% strains were carrying transferable drug resistant factor containing ampicillin, chloramphenicol, streptomycin and tetracycline. PMID- 8713244 TI - Role of transvaginal sonography in pelvic scan for female reproductive system. AB - Three hundred twenty-nine cases of early pregnancy and 116 pelvic pathology studied by transabdominal sonography were compared with 81 cases of early pregnancy and 88 cases of pelvic pathology subjected to transvaginal sonography. A high frequency vaginal probe, because of its close proximity with the target organ, produces remarkably sharp image. An accurate diagnosis is possible in great majority of cases within a short time. Vaginal sonography is done with an empty bladder. A close serial monitoring of ovarian follicles, endometrium and cervical mucus with transvaginal sonography offers an immense wealth of information about the structural and reproductive endocrinal status of the patient. Ovulation can be predicted in advance. Imaging of female reproductive system by transvaginal sonography is indispensable for any modern gynaecological care and for infertility assessment in particular. PMID- 8713245 TI - Histopathology of auricular appendages in rheumatic heart disease. AB - Forty-four left auricular appendages removed from patients presenting clinically as rheumatic heart disease (RHD) were subjected to histopathologic examination. The aim was to identify the changes so far reported in RHD. It was found that Aschoff's nodules, though specific for RHD, are much less frequent than the other changes reported. Viewing the histopathologic alterations in the perspective of clinical data is important in identifying the rheumatic process. PMID- 8713246 TI - Comparison of pleural fluid cytology and pleural biopsy in the evaluation of pleural effusion. AB - Pleural fluid cytology and pleural biopsy results were studied in 65 cases of pleural effusion. The efficacy of pleural biopsy in diagnosis of neoplastic and non-neoplastic pleural diseases was compared. Of the 24 cases with confirmatory evidence of cancer, 17(70.8%) has positive cytologic findings in pleural fluid, whereas pleural biopsy was diagnostic in only 13 cases (54.1%). For non-malignant pleural effusion in 41 cases 40(97.5%) has a definite diagnosis (tuberculous pleuritis, acute fibrinous pleuritis or hydatid cyst) which could be made by cytology while only 31(75.6%) out of 41 were diagnosed on pleural biopsy. The study indicates that cytologic evaluation of pleural fluid is more efficaceous in the diagnosis of malignant and non-malignant pleural disease than percutaneous pleural biopsy. PMID- 8713247 TI - Holistic concept of health. PMID- 8713248 TI - India's health - today and tomorrow. PMID- 8713249 TI - Leiomyoma of the jejunum presenting as peritonitis. PMID- 8713250 TI - Plexiform (multinodular) schwannoma in a child - a rare variant of schwannoma. PMID- 8713251 TI - Loudspeakers causing noise pollution. PMID- 8713252 TI - Guidelines for the diagnosis and treatment of dysentery. PMID- 8713253 TI - Parameatal cyst of penis. PMID- 8713254 TI - Lymphangioma of the parotid. PMID- 8713255 TI - Endomyometriosis. PMID- 8713256 TI - Secondary vesical calculus following translocated iucd in urinary bladder. PMID- 8713257 TI - Unilateral obstructive emphysema of the lung caused by mucus plugs. PMID- 8713258 TI - Buried rubber band: a cause for persistent penile discharge. PMID- 8713259 TI - Primary mucinous adencoarcinoma of the skin. PMID- 8713260 TI - Fibroma of the pinna. PMID- 8713261 TI - Hepatitis B e antigen in chronic HBV infection--a villain or a guide? PMID- 8713262 TI - Prevalence of hepatitis B e antigen in pregnant women and patients with liver disease. AB - Knowledge of HBeAg status is important in pregnant women to decide immunoprophylaxis to the infant and to decide therapy in patients with HBsAg positive chronic active hepatitis. Two thousand pregnant women were tested for HBsAg by the Hoechst IHA Cellognost Kit. Those who were HBsAg positive were tested for HBeAg by ELISA (Hoechst). One hundred consecutive HBsAg positive patients admitted in hospital were tested for HBeAg by Elisa (Abott). The prevalence of HBsAg and HBeAg in pregnant women was 5% (100/2000) and 12% (12/100) respectively. The prevalence of HBeAg in the 100 consecutive HBsAg positive patients with liver diseas was 30% (14/46) in acute viral hepatitis and 52% (28/54) in chronic liver disease. Routine screening of all pregnant women in the third trimester for HBsAg and immunoprophylaxis to infants of BHeAg positive mothers atleast is immediately recommended. PMID- 8713264 TI - Smoking and alcohol intake in a rural Indian population and correlation with hypertension and coronary heart disease prevalence. AB - 3148 persons (1982 males and 1166 females) aged more than 20 years in a cluster of three villages were examined. The overall prevalence of smoking was 51% in males (n = 1006) and 5% in females (n = 54). Among male smokers there were 26% light smokers (< or = 5 bidis/day), 51% moderate smokers (6-20/day) and 17% heavy smokers (> 20 day) and in females there were 54% light smokers, 41% moderate smokers and 5% heavy smokers. Smokers were less educated and had higher prevalence of work-related physical activity and alcohol intake. There was a higher prevalence of hypertension and of ECG Q-waves in male smokers. Regular alcohol intake was seen in 19% males (n = 377) and in 2% females (n = 26). Among males there were 43% light drinkers (< or = 28 gm ethanol/day), 32% moderate drinkers (28-56 gm ethanol/day) and 5% heavy drinkers (> 56 gm ethanol/day). Although this group had a higher prevalence of hypertension there was an insignificant difference in CHD prevalence and a significantly lower prevalence of ECG Q-waves. Subgroup analysis has also been performed taking non-smoker-non alcohol consuming group as controls. It was found that group which comprised of smokers-non-alcohol consumers had a significantly higher prevalence of hypertension and of ECG Q-waves. Alcohol intake-smoker group had a significantly higher prevalence of hypertension. The habits of smoking and alcohol consumption are widely prevalent among males in this rural community. Smoking and alcohol intake, both individually and collectively, are related to higher prevalence of hypertension as well as CHD. While the prevalence of hypertension is more among the alcohol consumers, smokers have a higher prevalence of CHD. PMID- 8713263 TI - Recurrent Guillain Barre' Syndrome: a clinical, electrophysiological and morphological study. AB - Of the 220 patients of acute idiopathic demyelinating polyneuritis (AIDP/GBS) seen over a seven year period, 15 patients (M:F:11:4) had a relapsing course (6.8%). Their ages ranged from 8 yrs to 70 yrs. They had 36 episodes at a variable interval of 3 months to 25 yrs. Relapse rate varied from one to four. Antecedent events were noted during 16 episodes in 9 patients but the triggering factors were varied. Clinical features of individual episodes were similar to the acute monophasic illness, although they differed inseverity from one episode to the other. Autonomic disturbances were rare. Albuminocytological dissociation was observed during 19 of the 24 episodes. Electrophysiological abnormalities were observed during 19 of the 24 episodes. Electrophysiological abnormalities were present in all and were comparable with patients of non-recurrent illness. Sural nerve biopsy in 3 patients showed evidence of demyelination, remyelination, Wallerian degeneration and myelin breakdown but none had features of inflammation. With the exception of one death, functional recovery was complete in the majority of patients, irrespective of the type of therapeutic intervention. Acute onset, frequent facial involvement, brief clinical course, near complete recovery and very long asymptomatic periods may distinguish these patients of acute relapsing demyelinating polyneuropathy (ARDP) from chronic relapsing demyelinating polyneuropathy. Relapses in GBS are however unpredictable and recurrent GBS is indistinguishable clinically, electrophysiologically and morphologically from the more frequently seen non-recurrent form of monophasic GB Syndrome. A biochemical or immunological marker may help in this distinction. PMID- 8713265 TI - Malaria--still a master killer? PMID- 8713266 TI - Neuromuscular changes in acute renal failure. PMID- 8713267 TI - Lipid peroxidation in acute aluminium phosphide poisoning. PMID- 8713268 TI - Drug induced emergencies. PMID- 8713269 TI - Newer calcium antagonists for treatment of systemic hypertension. PMID- 8713270 TI - Hemorheology in cerebrovascular accident: an overview. PMID- 8713271 TI - Dermatomyositis with malignancy. PMID- 8713272 TI - Double viral infection during pregnancy. PMID- 8713273 TI - The myopathic variety of arthrogryposis multiplex congenita. PMID- 8713274 TI - Basal ganglia calcification--the diversity within. PMID- 8713275 TI - Laurence-Moon-Biedl-Bardet syndrome with chorea. PMID- 8713276 TI - Priapism as a presentation of hematological malignancy. PMID- 8713277 TI - Lupoid sclerosis with raised anticardiolipin antibodies. PMID- 8713278 TI - Global aphasia without hemiparesis. PMID- 8713279 TI - ABO blood group distribution in kala-azar in Bihar, India. PMID- 8713280 TI - A case of symptomatic palatal myoclonus. PMID- 8713281 TI - Niemann Pick disease presenting as recurrent episodes of osteomyelitis. PMID- 8713282 TI - Anaemia and raised erythrocyte sedimentation rate. PMID- 8713283 TI - Three cases of acute porphyria-experience with heme therapy in one case. PMID- 8713284 TI - Rumination-treatment with cisapride. PMID- 8713285 TI - Myoglobinuric versus rhabdomyolytic ARF. PMID- 8713286 TI - Treatment of visceral leishmaniasis unresponsive to pentamidine with amphotericin B. PMID- 8713287 TI - HIV lymphadenopathy--a histopathological and immunomorphological study of 65 cases. AB - Lymph nodes from 65 HIV-infected persons were studied by morphological and immunomorphological methods. Early stages of HIV lymphadenopathy (follicular hyperplasia without fragmentation or follicular hyperplasia with severe fragmentation) were revealed in 92% of patients with PGL and the most advanced stadium of HIV-related lymphadenopathy (diffuse pattern) in 92% of AIDS patients. Striking B-cell activation and polyclonal proliferation, found in 19 lymph nodes (35% of advanced lymphadenopathy cases--follicular atrophy or diffuse pattern) justified separation of these cases as a particular subgroup of HIV lymphadenopathy. PMID- 8713288 TI - Environmental factors may regulate BCL2 associated lymphomagenesis: a very low incidence of BCL2-MBR translocation in Poland. AB - A characteristic feature of follicular non-Hodgkin's lymphomas (FL) is a chromosome translocation t(14;18)(q32;q21). In American patients the t(14;18) can be found in the large majority (approximately 70%) of FL and in a significant (10 40%) percentage of diffuse large cell lymphomas (DLCL). However there are reports suggesting that geographic or racial factors may regulate genesis of lymphomas with BCL2 abnormalities. Herein we present results of molecular analysis for t(14;18) in lymphomas in Poland. Analyses were performed on 37 cases of FL, 55 cases of diffuse lymphomas (DL) and 39 cases of Hodgkin's disease (HD). By Southern and polymerase chain reaction BCL2 translocation within major breakpoint region was found only in 3 of 37 FL, 1 of 55 DL and 2 of 39 HD. These results are the lowest reported ratio. We hypothesize that environmental factors may regulate BCL2 associated lymphomagenesis. PMID- 8713289 TI - The mast cells and an experimental gastric ulcer. Histochemical, ultrastructural and quantitative study in the rat. AB - The role of mucosal mast cells (MMC) in chronic non-allergic reactions of the gastric mucosa is unknown. The present study aimed to investigate the reaction of gastric MMC during healing of the acetic acid-induced gastric ulcer in the rat. Mast cells were stained with toluidine blue and alcian blue+safranin and their density and distribution were assessed. Morphometry of MMC and documentation of their contact with eosinophils were based on electron microscopy. In the normal non-injured rat oxyntic mucosa MMC are grouped in two populations: MMC1 concentrated near the glandular necks, and MMC2 in the basal lamina propria. There was a significant decrease in the number of MMC2 near the ulcer, whereas MMC1 lost their concentration in the neck zone. Also the MMC/connective tissue mast cells ratio was increased in the gastric wall adjacent to the ulcer. Eosinophils were commonly in close contact with MMC. Eosinophil cytoplasm adjacent to MMC was devoid of organelles, which were accumulated in the central cytoplasm. The significant redistribution of mast cell population as well as numerous close contacts between mucosal mast cells and eosinophils, taking place in the neighborhood of the chronic gastric ulcer, seem to be not only of morphological but also of functional significance. PMID- 8713290 TI - Leucine enkephalin level in rat hypothalamus, striatum, hippocampus and adrenal glands in experimental rat uremia. AB - The aim of the study was to analyze the effect of experimental uremia elicited in Wistar rats by 5/6 kidney resection on the leu-enkephalin level in hypothalamus, striatum, hippocampus and adrenal glands. We found, that in uremic rats leu enkephalin levels decreased in striatum and in adrenal glands. The level of leu enkephalin in adrenal glands was directly related to plasma creatinine. The weight of uremic rats was significantly lower than that of control rats. PMID- 8713291 TI - Controversies about the genetic model of colorectal tumorigenesis. AB - According to the genetic model, intestinal tumorigenesis is a result of the ordered in time inactivation of tumor suppressor genes and the activation of oncogenes. A tacit assumption is that both genes involved in the regulation of proliferation and growth factor-inducible genes, although inactivated, would not be changed during that process. The model requires that cancer cell population is homogenous, exists in a deterministic environment, and develops in a teleological manner. Meanwhile, tumorigenesis is rather a combination of both deterministic and stochastic molecular phenomena. Therefore, a novel notion of bifurcating point genes is defined as a generalization of the idea of tumor suppressor genes and oncogenes. Alternative stochastic mechanisms of tumorigenesis are discussed such as a decreased expression of intestinal-specific genes in cancer cells, most likely reflecting adaptation to survival within a heterogeneous, and non equilibrated cellular population. PMID- 8713292 TI - Quality control in cytopathology applied to screening for cervical carcinoma. AB - When screening for cervical carcinoma became one of the priorities of the European Community (1992), the "European Guidelines for Quality Assurance in Cervical Cancer Screening" proposed an organisation of the screening program. In the following years, Quality Assurance/Quality Control (QC) was organized in many countries. The whole staff of the laboratory has to participate in the organization of Internal QC programs: errors must be detected in all categories of smears and the same terminology has to be adopted. External QC is well accepted when organized by the pathologists themselves. The French program is presented. In the near future, pathologists should take into account the evolution of QC, and the role of modern methods such as telecytopathology and computer-assisted screening devices. PMID- 8713293 TI - Advances in cancer research: report from the eighty-sixth annual meeting of the American Association for Cancer Research. AB - A meeting of the American Association for Cancer Research was held in Toronto, Canada on March 18-22, 1995 to discuss advances in cancer research. Growing interest was shown in the molecular mechanisms of prostate cancer. Both breast cancer and colorectal cancer continued to be major scientific topics of interest. Research into the genetics of cancer was dominated by the issue of genomic instability due to failures in the mismatch DNA repair gene system of cells. Instability appeared to represent the key mechanism for the generation of mutations. Another common theme was the identification of putative tumor suppressor and metastasis loci using loss of heterozygosity and cell chromosome fusion approaches. A few novel sequences with a potential suppressive function were reported. Also, relationships between growth factor receptors, tumor suppressor genes, oncogenes, and proteins controlling cell cycle were analyzed. Progress in gene therapy and chemoprevention of malignant tumors was frequently discussed. PMID- 8713294 TI - Malignant melanoma of the larynx. AB - A case of laryngeal melanoma is presented. Difficulties in the differential diagnosis between primary and metastatic tumours are discussed. PMID- 8713295 TI - [The ultrastructural aspects of the stress-limiting action of a pulsed electrical current on the hypothalamic neuronal subpopulations that initiate the body's stress reactions]. AB - Electron-microscopic investigations of neuron subpopulations in paraventricular nuclei of rat hypothalamus demonstrated ultrastructural positive changes due to exposure of the stressed animals to impulse electric current (IEC). The latter is able to control synthesis and secretion of stress-releasing-hormones produced by neurons of hypothalamic medial small-cell and posterior large-cell subnuclei. There were indications of deposition of stress-initiating hormones in neuron bodies evidencing stress-limiting effect of IEC. There were also signs of elevated resistance of ultrastructures, endoplasmic reticulum, in particular, to stress-related damage. PMID- 8713296 TI - [The mechanism of the action of electromagnetic radiation in the nano- and millimeter ranges]. AB - It is suggested that there exists not only resonance but also interference interaction between laser radiation, millimetric electromagnetic radiation and dissipative systems. A noticeable role here is played by opiate receptors and their ligands determining cytoprotective, stress-limiting, immunomodulating and other effects of EHF therapy. PMID- 8713297 TI - [The function of the kallikrein-kinin system and the activity of proteinase inhibitors during physical loading and infrared laser irradiation]. AB - Experiments on 40 male rats were conducted to examine kallikrein-kinin system (KKS) and activity of proteinase activity under stress, impulse infrared laser radiation and their sequence. Infra-red laser radiation following stress prevents activation of KKS provoked by exercise through enhancing antiproteinase potential of blood serum. Thymic mass and activity of thymocyte genome were on the increase. PMID- 8713298 TI - [The effect of iodobromide baths on the physical work capacity and extrasystole in patients with ischemic heart disease and stable stenocardia]. AB - 68 patients with CHD and stable angina pectoris class 1-2 in the presence of extrasystole were examined (spiro-bicycle ergometry, Holter ECG monitoring) before and after a course of general iodobromide baths (38 patients) or general fresh-water baths (30 patients). General iodobromide baths exerted marked antiarrhythmic effect in coronary heart disease with stable angina pectoris, the occurrence of ventricular, supraventricular extrasystoles reduced at least by half. Painful and silent myocardial ischemia decreased. Due to training effect of iodobromide balneotherapy muscular performance and coronary heart reserves improved. PMID- 8713299 TI - [The use of laser therapy and physical exercises in myocardial infarct patients at the hospital stage]. AB - 89 postmyocardial infarction patients were exposed to low-intensity laser radiation combined with muscular training. The treatment improved clinical condition of the patients, myocardial contractility, exercise tolerance. PMID- 8713300 TI - [The efficacy of using an electromagnetic field of extremely high frequency (54 78 GHz) in treating patients with chronic nonspecific lung disease]. AB - After a trial of the therapeutic complex including extra high frequency electromagnetic field in 154 patients with chronic bronchitis and bronchial asthma high efficacy of EHF-therapy was stated in the above diseases. PMID- 8713301 TI - [The effect of the ultraphonophoesis of deresinated naphthalan in patients with a history of exudative pleurisy]. AB - Clinicoimmunological comparisons suggest the conclusion on positive effect of deresinated naphthalan phonophoresis when given early in the course of inpatient combined treatment of exudative pleurisy. The response was noted in the trend of clinical and immunological characteristics of T- and B-cell immunity. PMID- 8713303 TI - [The use of interference currents in the rehabilitative treatment of patients with fractures of the leg bones]. AB - Interference currents used in early treatment of patients with crural fractures accelarated recovery of the limb functions as a result of stimulation of involuntary muscular activity, reduced tissue adema and increased mobility of the ankle joint. PMID- 8713302 TI - [Experience with physiotherapy in a posttraumatic neurodystrophic syndrome of the extremities]. AB - Combined pathogenetic treatment including physiotherapy, drugs, massage, exercise was tested in 98 patients with posttraumatic neurodystrophic syndrome in phase II and III. The responses achieved made it possible to recommend the above regimen for wide application in rehabilitation centers and clinics. PMID- 8713304 TI - [A trial of the clinical use of a new method of electrosleep]. AB - A novel device and technique of interference electric sleep therapy have been developed and tried in 125 children and 324 adults with different diseases. The highest clinical response, especially in children, was achieved on Elson unit. PMID- 8713305 TI - [The use of an electrosleep method for restoring the performance capacity and relieving the psychoemotional stress of athletes in cyclic sports]. AB - 44 boat-racing sportsmen entered the study which aimed at elucidation of electric sleep action on restoration of muscular performance and relief of psychoemotional stress. The electric sleep procedures were found to stimulate regeneration processes, improve cardiovascular function and myocardial contractility, normalise arterial and capillary tonicity, to potentiate muscular performance and aerobic power of the body. PMID- 8713306 TI - [The assessment at a sanatorium of the mental status of children who were victims of the Chernobyl catastrophe]. AB - Psychic disorders are not infrequent findings in children who survived Chernobyl accident. This fact necessitates special psychotherapeutic, psychocorrective and psychoprophylactic treatment which should be added to standard therapeutic care in sanatoria. PMID- 8713307 TI - [Legal standards for the rational utilization of natural curative resources and their protection from pollution]. PMID- 8713308 TI - [The assessment of the biological status of sapropels by their enzymatic activity]. PMID- 8713309 TI - [The problems of physical therapy]. PMID- 8713310 TI - [The effect of adaptation to hypoxia on the results of the health resort treatment of children with bronchial asthma]. PMID- 8713311 TI - [Laser therapy of patients with climacteric disorders]. PMID- 8713313 TI - [Medical support of sports for the disabled]. PMID- 8713312 TI - [Conceptual approaches to the problems of the seasonal health promotion of children and adolescents]. PMID- 8713314 TI - [The therapeutic use of physical factors in the combined therapy of patients with psoriatic arthritis]. PMID- 8713315 TI - [Level of glucose in blood, gas exchange, and hematologic indicators of rats with experimental diabetes mellitus after carotid glomectomy]. PMID- 8713316 TI - [Comparative analysis of the effects of prolonged peripheral and intracerebral administration of beta-endorphin]. PMID- 8713317 TI - [Role of ATP as a mediator of the sympathetic nervous system in smoothing out rapid changes in arterial pressure]. PMID- 8713319 TI - [Ulcer damage of the stomach in August and Wistar rats under acute emotional stress]. PMID- 8713318 TI - [Effects of L-arginine and its functional antagonist N-nitro-L-arginine on behavior]. PMID- 8713320 TI - [Free radical damage to the liver during transplantation in oxidative stress]. PMID- 8713321 TI - [Correlation between blood supply and contraction of myocardial segments from intact and ischemic hearts]. PMID- 8713322 TI - [Comparative study of the dynamics of hemodynamic effects of immobilization stress in normotensive, stress-sensitive, and spontaneously hypertensive rats]. PMID- 8713323 TI - [Adaptation to hypoxia, as opposed to adaptation to stress, does not protect the isolated heart from reperfusion after total ischemia. A nuclear magnetic resonance study]. PMID- 8713324 TI - [The effect of methylcholanthrene and sovol induction on the functional activity of rat blood neutrophils in vivo]. PMID- 8713325 TI - [Protective effect of M- and N-cholinergic receptor blockaders during acute ammonia poisoning]. PMID- 8713326 TI - [Change in activity of enkephalinase from various structures of the rat brain during administration of naloxone and in morphine tolerance]. PMID- 8713327 TI - [Functional status of endothelium and production of nitric oxide in the rat, adapted to periodic hypoxia]. PMID- 8713328 TI - [Effect of beta-casomorphine-7 on different types of training of white rats]. PMID- 8713330 TI - [Features of the course of the candida infection process during combined effect of medications]. PMID- 8713329 TI - [Comparative study of the effect of amiridine on biological membranes]. PMID- 8713331 TI - [Stimulating effect of a polypeptide substance (Cuban stimulator) on hemopoiesis and formation of humoral immunity to murine thymus-dependent antigen]. PMID- 8713332 TI - [Cytotoxicity of carotene compounds on human cultured epidermal carcinoma cells]. PMID- 8713333 TI - [Lifetime quantitative assessment of intracellular distribution of doxorubicin in tumor cells]. PMID- 8713334 TI - [Experience in use of biologically active substances from human fetal tissue in treating oncological diseases]. PMID- 8713335 TI - [Role of Ehrlich ascites carcinoma adrenergic receptors in regulating proliferative processes in it by adrenergic ligands]. PMID- 8713336 TI - [Collagen-based biologically active coatings]. PMID- 8713337 TI - [Design of a Yersinia pestis strain with increased protection]. PMID- 8713338 TI - [Morphological changes in erythrocytes exposed to perfluorodecalin and oxygenation]. PMID- 8713339 TI - [Ultrastructural-cytochemical analysis of cardiomyocytes from young and old rats during generalized overheating]. PMID- 8713340 TI - [Isolation of soluble immune complex in biopsied tissues]. PMID- 8713341 TI - [Changes in cardiohemodynamic indicators during use of verapamil for preventing and treating hypertensive complications of pregnancy]. PMID- 8713342 TI - [Metastasis of induced uterine sarcoma]. PMID- 8713343 TI - [Possible circulatory paths of endogenous human retroviruses, similar to murine mammary cancer virus]. PMID- 8713344 TI - [Comparison of the level of endogenous paramagnetics in blood serum in normal conditions and pathological states by proton magnetic relaxation]. PMID- 8713345 TI - [Radioimmunological determination of glucagon in Bulgarian insulin substances]. AB - The contents of glucagon in insulin substances ("Pharmachim" Ltd.) is determined in accordance to the needs of pharmaceutical production. The middle quantitative values and standart deviation are specified. Nevertheless that monocomponent and bovine conventional insulin substances pertain to groups of different purity they exibit the highest purity. Bovine conventional insulins exibit the greatest variability, while monocomponent insulins are destinctively homogenous. Single peak insulin substances exhibit less purity, but this group is more homogenous. The middle quantitative value is within the admissible limitations. Porcine conventional insulins have high contents of glucagon as well as high values of standard deviation. PMID- 8713346 TI - [The reactivity of canine colonic and ileal smooth muscle to carbachol (Jestryl)]. AB - Experiments were carried out on longitudinal and circular smooth-muscle strips isolated from the ileum and colon of healthy dogs (controls) and of dogs with local peritonitis. The changes in the contractile responses of the strips to carbachol administered cumulatively were studied, and the EC50 and pD2 values were calculated from the dose-response curves. For the colonic longitudinal strips the EC50 was 6 x 10(-8)M and the pD2 was 7.58. The pD2 for the ileal longitudinal strips was 7.83. The pD2 value for the circular strips from the ileum and colon was 7.19 and 8.87 respectively. In the dogs with local peritonitis, the sensitivity of the longitudinal strips to carbachol was higher (the pD2 for the ileum was 7.86 and for the colon-7.21) as compared to the circular strips (the pD2 for the ileum was 6.22 and for the colon-6.65). The affinity of the cholinoreceptors in the ileal circular strips to carbachol was decreased as compared to controls. PMID- 8713347 TI - [The determination of the somatostatin content in insulin substances]. AB - 30 different lots (batches) of insulin substances ("Pharmachim" Ltd.) are analysed and the contents of somatostatin is established. The middle quantitative values and the standard deviations are specific. Though that monocomponent and bovine conventional insulin substances pertain to groups of different purity they exhibit highest purity but also greatest variability, while single-peak and porcine conventional insulins show less purity but these groups are more homogenous. PMID- 8713348 TI - [The morphological changes in the liver in experimentally induced hypercholesterolemia]. AB - The authors examined the liver of 20 rabbits "Newzeland white" and "Chinchila", undergone atherogenic diet with cholesterol (0.3-0.5 g/kg body weight) for 30 and 60 days. They determined cholesterol, lipids, triglicerides and beta-lipoproteins in the serum. The morphologic study revealed accumulation of cholesterol in the hepatocytes, fibrosis, proliferation of the bile ducts and signs of hepatic cirrhosis. PMID- 8713349 TI - [Morphological studies of the lungs in experimental burns]. AB - The authors studied light- and electronmicroscopically the lungs of 20 rats after experimental burns in closed space. Twenty one histological parameters were evaluated. Two mechanisms of quickly developed fatal outcome were established- asphyxia and acute injury of shock lung. Electronmicroscopically there was an increased secretion of surface active materials and degenerative changes in the alveolar epithelium and capillary endothelium. Carbon particles found in the upper respiratory tract were considered as signs of vitality. PMID- 8713350 TI - [The functional and morphological changes in the liver and kidneys of white rats treated with aluminum]. AB - A total of 126 white male Wistar rats under subacute conditions were studied. Of them, 76 were perorally treated with AlCl3 in a dose of 3 mg/kg b.w. daily for 40 days. Activities of acid phosphatase (AP), succinate dehydrogenase (SDH), adenosine triphosphatase (ATP-ase) and the contents of glycogen, glucoproteins and RNA were dynamically followed-up in the liver and kidney. Serum activities of ASAT and ALAT were estimated by Borhringer's tests. Morphological changes were histochemically investigated. There was an initial elevation followed by reduction mostly manifested for SDH and ATP-ase and an increase of AP at the end of trial. Histochemical data argued for disorders of protein and carbohydrate metabolism. Morphological alterations more outlined in the liver became more severe until the end of the experiment. PMID- 8713351 TI - [The polarization microscopy characteristics of collagen in dura mater transplants]. AB - In order to establish the polarizing microscopical characteristics of collagen in preserved dura mater transplants and in controls, a study was performed. Attempt was made for an intrpretation of velocity of collagen rehydratation by means of data of total anisotropy. The results show a faster rehydratation of collagen fibrils in control (unpreserved) dura mater. Changes in the velocity of rehydratation of collagen fibrils in lyophilized dura mater were connected with structural changes during lyophilization. The most appropriate period of time for rehydratation of lyophilized dura mater transplant was determined from the results of the polarizing microscopical study. PMID- 8713352 TI - N-methyl-D-aspartate-induced 45Ca2+ release from pre-labelled adult rat hippocampus in vivo. AB - We report the results of microdialysis experiments investigating the NMDA-induced release of intracellular Ca2+ in different brain regions. Microdialysis probes were implanted stereotaxically into the striatum, thalamus and hippocampus dentate gyrus (DG) of adult rats. Dialysates were analysed for alterations in the concentration of ionized Ca2+ in an initially calcium-free medium and for changes in 45Ca efflux from the pre-labelled endogenous Ca2+ pools. The application of 5 mM of NMDA to the dialysis medium for 20 min in the striatum, resulted in increases in Ca2+ and 45Ca concentrations by 25% and 35% respectively. After NMDA perfusion in the hippocampus DG and in the thalamus, decreases in the Ca2+ concentration to 65.6% and 38.6% of the basal level respectively, were accompanied by increases in 45Ca efflux, exceeding 1,500% of the basal level in the hippocampus. Cell swelling, and the corresponding reduction of the extracellular space volume was insufficient to explain the huge increase in 45Ca efflux. Thus, our experiments demonstrated that in vivo in the rat hippocampus DG, NMDA induces the release of 45Ca to the extracellular space from unidentified intracellular calcium stores. PMID- 8713353 TI - Temporal dynamics and regional distribution of [14C]serine uptake into mouse brain. AB - In order to examine the uptake of L-serine into brain structures and brain metabolic compartments, L-[U-14C]serine was injected into tail vein of mice. The uptake was examined 30 min, 90 min, 3 h and 5 h after injection by both quantitative autoradiography of coronal brain sections and by biochemical analysis. Brain radioactivity was extracted and partitioned into protein associated pellets, metabolites soluble in aqueous phase and lipids soluble in the organic phase. Most of the radioactivity was found in the aqueous phase, about 10% was incorporated into lipids. Among phospholipids the highest label was found in phosphatidylserine, then in phosphatidylethanolamine and in phosphatidylcholine, it amounted to 52%, 30% and 18% of label by 90 min after injection, respectively. The brain distribution of L-serine uptake resembled that described for strychnine-insensitive [3H]glycine binding, with cortical structures being preferentially labelled. PMID- 8713354 TI - Participation of different 5-HT receptors in the memory process in rats and its modulation by the serotonin depletor p-chlorophenylalanine. AB - The memory effects of agonists and antagonists of some serotonin (5-HT) receptor subtypes were examined in experiments on rats using an active avoidance method (shuttle-box). The 5-HT receptor antagonists NAN 190 (1 mg/kg i.p.) and pindolol (6 mg/kg i.p.) improved some indices for memory; the 5-HT2 and 5-HT3 receptor antagonists ritanserin (1 mg/kg i.p.) and ondansetron (0.1 mg/kg i.p.) exerted a favourable effect on the mastering of active avoidance performance. The tryptophan hydroxylase inhibitor para-chlorophenylalanine (300 mg/kg i.p.) alone produced no significant changes in the indices for retention of learned behaviour but in combination with the 5-HT-receptor agonists and antagonists influenced some of their effects. The results obtained show different participation of 5 HT1A, 5-HT2 and 5-HT3 receptors in the mechanisms of the memory process; the nature of this involvement is modulated by the brain level of serotonin. PMID- 8713355 TI - Spermine protects in vivo the antioxidant enzymes in transiently hypoperfused rat brain. AB - The antioxidant enzymatic system in brain hypoperfusion/reperfusion model in rats after spermine administration was evaluated. Incomplete cerebral ischemia/reperfusion induced by temporal occlusion of common carotid arteries caused a decrease in the activities of superoxide dismutase and glutathione reductase as well as total and free sulfhydryl groups, while thiobarbituric acid reactive substances became elevated. Administration of spermine after the reperfusion led to restoring all above parameters to normal values. Protective effect of spermine in transiently hypoperfused and subsequently reperfused rat brain is briefly discussed. PMID- 8713356 TI - Survival and maturation of heterotopic fetal brain stem xenografts after treatment with 2-chlorodeoxyadenosine and cyclosporine A. AB - Brain stem halves from fetal rabbits were transplanted to the caudate nucleus area of adult rats. The animals were treated postoperatively with cyclosporine A (CsA) and 2-chlorodeoxyadenosine (CdA) for three days, and with CdA alone for the next 13 days. The treatment started at the day of implantation, and in some animals it was repeated starting at day 36 after grafting (at the time when signs of a light inflammatory reaction appeared in some grafts). Grafts survived and matured histologically, and no signs of acute rejection were observed up to the 90th day. In some grafts we recorded phasic neuronal activities similar to the respiratory-related neural activities characteristic for the adult brain stem. Immunosuppressive with CdA and CsA deserves further evaluation in fetal brain grafting. PMID- 8713357 TI - Social interactions in cats: regional brain monoamine distribution in dominant and submissive cats. AB - Regional brain concentration of monoamines (NA, DA and 5-HT) and their metabolites (MHPG, DOPAC, HVA and 5-HIAA) were measured in dominant and submissive cats in a predatory competition test and in predatory behaviour of single cats. A submissive position in predatory hierarchy produced an increase in concentration of NA in the hypothalamus, a decrease of DA, 5-HT, MHPG and MHPG/NA ratio in the hippocampus and a decrease of 5-HT in the prefrontal cortex in comparison with dominant cats. In the predatory test the social situation increased the concentration of DA and its metabolites in the hypothalamus of the submissive and dominant cats as compared to single cats. Additionally, a diminution of NA and MHPG concentration in the midbrain in dominant cats compared to single animals in the predatory situation was observed. The data obtained demonstrate a considerable differentiation in the effects of predatory aggression and predatory behaviour in a social situation on brain distribution of monoamines. The results indicate that predatory behaviour in single and paired cats is regulated by different catecholamine mechanisms. PMID- 8713358 TI - Effects of repeated systemic penicillin injections on nonconvulsive and convulsive epileptic seizures in the rat. AB - Changes in the spontaneous and induced epileptic activity in the course of repetitive systemic i.p. administration of crystalline penicillin (Pc) were examined in imp-DAK rats. In all the rats used, nonconvulsive seizures characterized by bursts of spike and wave discharges (SWD) in the neocortex occurred spontaneously. A single i.p. injection of Pc at doses of 1,000,000, 1,500,000 or 2,000,000 IU/kg resulted in a transient increase in SWD activity. For the latter two doses, Pc injections also induced convulsive activity, i.e. single spikes and trains of spikes accompanied by myoclonies. When 1,500,000 IU/kg of Pc was administered repeatedly (six injections, one every 48 h), the amount of the convulsive activity induced by successive injections decreased but the increase in the number of SWD bursts became more pronounced. This result gives rise to some questions about the development of tolerance to the epileptogenic Pc effects in the course of repeated administration of this antibiotic. PMID- 8713360 TI - Perseverative errors and reversal of a visual discrimination following basal forebrain lesions in the rat. AB - Rats with electrolytic lesions of the nucleus basalis magnocellularis were compared to sham-lesioned rats in the retention of a continuously reinforced lever-pressing response and in the acquisition and reversal of a visual discrimination task. The muscarinic agonist pilocarpine, in conjunction with the peripheral muscarinic antagonist methyl-scopolamine, was administered in three doses to subsets of each group during acquisition. The lesion interfered with retention of the lever-pressing response. It did not affect the rate of acquisition of the visual discrimination, but facilitated reversal, and increased the number of perseverative errors made by the rats. Pilocarpine's only notable effect was to increase the latency to respond. PMID- 8713359 TI - Sensory conditioning and sensory stimulation do not affect GABAA receptor binding in the barrel field of mice. AB - The whisker-to-barrel system of adult mice was used in a study on the effects of short-lasting tactile stimulation and sensory conditioning training on GABAA receptor binding in the barrel field of somatosensory cortex. In vitro receptor binding autoradiography was used to examine the pattern and intensity of [3H]muscimol binding to GABAA receptors. A well-defined pattern of GABAA receptors in the barrel field remained unaffected after both procedures used. Also, no differences in intensity of GABAA receptor binding were observed. These results suggest that GABAA receptors are not involved in the plastic changes developing during sensory conditioning training. PMID- 8713361 TI - Two components of long-term memory. AB - The existence of two independent components of long-term memory has been demonstrated by the authors. The evidence has been derived from the authors' findings related to the optimum spacing of repetitions in paired-associate learning. The two components are sufficient to explain the optimum spacing of repetitions as well as the spacing effect. Although the molecular counterparts of the two components of memory are not known, the authors provide a collection of guidelines that might facilitate identification of such counterparts. PMID- 8713362 TI - PC based EEG mapping system. AB - A PC based program has been designed for analysis of EEG data using brain electrical activity mapping technique. It operates under control of MS-DOS operating system and can be used as a stand alone program or incorporated into a digital EEG system. The program can apply different map interpolation algorithms from nearest neighbours methods to spherical spline functions. It performs analyses in the time and frequency domain. It applies different mapping parameters like amplitude, spectral values and a new one--first time derivative of amplitude. The program is being successfully used in clinical tests of epilepsy and Parkinson disease in our laboratory. PMID- 8713363 TI - [Coronary assessment and surgery for aortic aneurysm: a pragmatic attitude]. AB - High prevalence of coronary artery disease in patients with AAA leads to a high rate of peri-operative cardiac complications. Coronary insufficiency is thus the cause of 40 to 60% of post-operative deaths after aortic surgery. Demonstration of coronary insufficiency depends on the clinical history, electrocardiographic evidence, non-invasive examinations and coronarography. Diagnosis is based on non invasive tests, and of primary importance exercise tests, which have a high sensitivity. Specificity for predicting post-operative cardiac complications remains low but can be improved by combining with other tests (for example exercise test and Holter recording) or with other clinical parameters. Coronarography provides a precise map of the coronary status but gives little information on functional impairment of encountered lesions. Finally, besides the cost and a certain degree of morbidity, coronarography increases the number of indications for revascularizations with the inconvenience of its intrinsic mortality and also retards the operation increasing the risk of rupture. The evaluation of cardiac risk before surgery must be based on correct use of non invasive tests, limiting coronarography to cases with frankly positive tests. PMID- 8713364 TI - [Coronary assessment and surgery for aortic aneurysm: systematic coronarography]. AB - Epidemiologic studies have shown that patients with peripheral vascular disease have a high risk of coronary heart disease. The vascular surgeon must therefore search for latent lesions which might lead to decompensation during or after the surgical procedure. Coronarography can be proposed in symptomatic patients or those with a past history of coronary heart disease, but such an indication is questionable for asymptomatic patients. Non-invasive exploration could theoretically provide information of the coronary risk but are not always possible or reliable in patients with peripheral vascular disease. A practical attitude can be established by analyzing the lesions on the basis of large international series. Cardiac exploration evaluating ischaemia can be proposed after the initial work-up to establish management strategy for intermediary cases. PMID- 8713365 TI - [Blood salvage: decisive progress in vascular surgery]. AB - There are two different methods of autotransfusion during an operation; one without blood washing and the other with wash red cells obtained after the filtering and concentration of the recovered blood. The first method has the advantage of being simple and cheap while recovering 1500-2000 ml of blood. Nevertheless it is insufficient in the case of an important or rapid bleeding. The weight of haemoglobin in the recovered blood is relatively low and the hemolysis is without clinical consequences. The rate of the coagulation factors is reduced. The autotransfusion with wash red cells requires a more important investment at the beginning. But the washing process eliminates the cell micro aggregates and nearly all of the substances in the recovered blood. Therefore concentrated units of red cells may be obtained with hematocrits between 45 and 65%. Autotransfusion with washing offers a greater security when the bleeding is important or violent. The autotransfusion is useful for a bleeding between 1 and 3 litres. The average volume recovered corresponds to 2.5 pockets of blood extract, thus covering the costs for the kits. If the bleeding exceeds 1.5 times the blood volume, blood extract and plasma will be required in addition. The risk of homologue blood transfusion can only increase the development of autotransfusion techniques and particularly the recovery of blood during the operation process. PMID- 8713366 TI - [Blood salvage: a conditional safety]. AB - Recovery of shed blood is part of the allogenic blood saving policy of particular importance even though the risk of viral infection via transfusion has been considerably reduced. Blood transfusion requirements in vascular surgery are discussed together with alternatives to allogenic transfusion. Differed withdrawal of autologous blood can involve pre-operative autologous plasmapheresis and cytapheresis. Per-operative haemodilution is another variant of pre-operative isovolemic haemodilution and erythrocytapheresis. Recovery of shed blood can be done with or without lavage. Technical and pharmacologic measurements complete the method. Simultaneous use of different techniques can be useful. Recovery is particularly interesting in highly haemorhagic vascular procedures or those which must be done quite rapidly. Care must be taken to avoid the "recovery syndrome". Improvement in material will assure safety. PMID- 8713367 TI - [Pros and cons of use of collagenated arterial prostheses]. AB - Use of bovine collagen for collagen arterial grafts raises the problem of possible contamination with the agents causing subacute spongiform encephalopathy. The safety of these products is the confirmed by the manufacturers and the risk of contamination is practically zero. A prospective non-randomized study conducted from January 1991 to December 1992 compared non impregnated arterial stents with collagen arterial stents. Among the parameters studied, only the amount of blood transfused was significantly different. Collagen arterial grafts generated less blood loss. PMID- 8713368 TI - [Impregnated polyester prostheses: a theoretical advantage]. AB - Leak-proof polyester grafts impregnated with collagen, gelatine or albumin are routinely used in vascular surgery. Theoretically, there are several advantages: no need for pre-coagulation, reduced operation time and graft manipulation, less blood loss during the operative period, better healing with a potentially greater resistance to infection. We analyzed all the publications in scientific journals to verify whether these theoretical advantages are validated by clinical trials. There have been 12 non-comparative series reporting favourable results but the lack of control groups makes it difficult to draw conclusions. Five comparative series with random assignment of patients have been reported. None of these series showed a substantial benefit from impregnation. Thus the use of an impregnated polyester graft would not appear to be justified for routine standard aorto-iliac surgery. Conversely, although no evidence has been provided by a comparative study, the major risk of haemorrhage in procedures such as extra corporal circulation, the thoracic or abdominal aorta and in patients with a coagulation disorder, impregnated graft probably constitute an important progress. PMID- 8713369 TI - [Treatment of abdominal aortic aneurysm: importance of the aorto-aortic tube?]. AB - We compared retrospectively the immediate and mid-term outcome in 203 operations for AAA performed at the Saint-Michel Hospital in the vascular surgery department. Two groups of patients were treated either with a tube graft or with a Dacron bifurcated graft. In our experience, the tube was prefered when there were no haemodynamically significant stenosis or aneurysmal dilatations in the iliac arteries. Tube implantation was shorter and caused less haemorrhage (p < 0.01), but post-operative mortality and morbidity was not significantly different. Patients with bifurcated graft had late vascular complications requiring reoperation more often. Most were anastomotic false-aneurysms. Three patients with a tube developed iliac lesions requiring an aorto-femoral or aorto bifemoral graft. Cure of an AAA with a tube graft is the choice treatment in patients free of severe iliac lesions or an aneurysmal bifurcation. The risk of subsequent iliac lesions requiring reoperation is low. PMID- 8713370 TI - [Treatment of abdominal aortic aneurysms: importance of the bifurcated prosthesis]. AB - Despite the increasing use of tube grafts to treat aortic aneurysms, bifurcated prostheses remain the most frequent solution. Advocates of the tube graft emphasize faster positioning and lower operative morbidity and mortality rates. However, the condition of the aortic orifice (where atheromatous lesions are maximal) and the aneurysmal or occlusive iliac disease frequently associated with aortic aneurysms usually require use of a bifurcation prosthesis for complete treatment of aortoiliac lesions. PMID- 8713371 TI - [Infrarenal aneurysm: traditional surgery]. AB - Abdominal aortic aneurysm surgery has been widely performed for more than forty years. Indications, contraindications, mortality-morbidity are clearly known. Surgical results are grossly reproducible whatever the centre. This classical technique of graft inclusion is superior to recent endovascular technique in terms of practicability and reproducibility. Classical technique seems safer for prevention of secondary aneurysms, for simultaneous treatment of abdominal non vascular lesions, for assessment and cure of associated renovascular abnormalities, and for avoiding distal emboli and colorectal ischemia. PMID- 8713372 TI - [Morbidity and mortality after elective surgery of complicated aneurysms]. AB - The post-operative mortality and morbidity after elective surgery of a complicated (difficult) infrarenal abdominal aortic aneurysm are reported. The authors compare a group of 126 patients with a simple uncomplicated aneurysm to a group of 126 patients with a complicated aneurysm. Mortality (3.2%) is similar in both groups but the morbidity is very high in the group of patients with difficult aneurysms. The most frequent complication was renal insufficiency determined by preoperative renal insufficiency. The length of the hospital stay was much longer for the patients after a surgery for a difficult aneurysm. A preoperative correct diagnosis is always possible and the surgical technique varies according the type of anomaly. Even elective surgery of a complicated aneurysm remains a challenge. PMID- 8713373 TI - [Medical treatment of arm embolisms]. AB - Emboli in the upper limb occur much less often than in other territories. They usually cause subacute ischaemia due to the importance of the arterial anastomotic network. Medical treatment is thus often sufficient. An analysis of the data in the literature show that medical treatment can give results comparable to surgery. Medical management should only be proposed as a complementary treatment when acute sensitivomotor acute ischaemia threatens the upper limb since priority must be placed on immediate restoration of the arterial flow. PMID- 8713374 TI - [Leg embolisms: treatment via surgical embolectomy]. AB - Arterial thrombembolectomy with the Fogarty balloon catheter was the standard therapy until now. We present fifty three patients who underwent 63 embolectomies. The rate of leg salvage was 95%, mortality was 18.8%. There is no controversy about the use of the Fogarty balloon catheter in the iliofemoral segment, particularly in patients with acute ischemia. In popliteal and tibial embolic occlusions, associated intraoperative fibrinolytic therapy obtained good results. Percutaneous thrombo aspiration was a complementary technique for few of distal embolic occlusions. PMID- 8713375 TI - [Leg embolisms: treatment via percutaneous thrombo-aspiration]. AB - PURPOSE: Benefits of the local thrombolytic therapy in the treatment of acute peripheral arterial ischemia have been proved (13), but the place of thrombo aspiration in this percutaneous treatment is not well defined. This study evaluates the results of thromboaspiration with or without local thrombolysis for the treatment of arterial embolism associated with a lower limb arteriopathy. METHODS: From May 1991 to May 1992, ten arterial embolisms (below the common femoral artery) associated with a lower limb arteriopathy have been treated by thromboaspiration with or without local thrombolysis. RESULTS: Nine immediate successes have been obtained. One failure has been operated on with success. On the third day, an occlusion of the popliteal artery occured and a femoroperoneal bypass was performed. But the occlusion of the bypass obligated to an amputation above the knee. This patient died one month later (hospital mortality 10%). During the follow-up (2 months), other procedures were patent. CONCLUSIONS: Thromboaspiration with or without thrombolysis is efficient for the treatment of arterial embolism associated with a lower limb arteriopathy. PMID- 8713376 TI - [Acute carotid occlusion: thrombolytic treatment]. AB - The occurrence of a neurologic deficit at the time of an acute obstruction of the internal carotid does not equate with neurons death. The size of the residual infarct depend on the duration and the depth of ischemia. The goal of fibrinolytic therapy is to obtain a fast reperfusin of the ischemic areas to limit the size of the residual infarct. The risk of reperfusion depend on the depth of the blood-brain barrier ischemia. The indications of reperfusion in emergency settings are based on pretherapeutic CTscan and angiographic assessment with cerebral digitalized parenchymography. Between 1984 and 1994, 100 ischemic strokes have been treated on emergency by local intra-arterial thrombolysis. The results depend on the condition of lenticulostriate arteries: --when the lenticulostriate arteries are not involved in the occlusion, arterial thrombolysis is very efficient (75% good results; 0% bad results) and has been performed up to the 12th hour. --when the lenticulostriate arteries are involved, the results are not as good (58% good results; 23% bad results); the hemorrhagic risk has dramatically dropped in this group when the decision was taken to do not treat the patients after the 5th hour (16.7% to 2.3%). There has been 7 deaths, 6 were due to non efficient revascularization of the parenchyma with vasogenic oedema. In conclusion, we think that ischemic stroke is an emergency; the cerebral digitalized parenchymography appears to be a major diagnostic and prognostic tool; intra-arterial thrombolysis is a very efficient technique when used at the right site and time. PMID- 8713377 TI - -Emergency surgical treatment of acute carotid occlusion-. AB - Conservative medical treatment of acute occlusion of the extracranial internal carotid artery usually gives mediocre results. When a major neurological deficit is involved, mortality can reach 16 to 55%, morbidity due to definitive deficit 40 to 69% and cure only 2 to 12%. It is thus logical to attempt revascularization as an emergency procedure. In situ intraarterial fibrinolysis is appropriate for acute occlusion in the intracranial territory of the internal carotid involving severe neurological deficits but surgery is more adapted and safer for acute occlusion of the extra-cranial internal carotid. In a personal series of 8 patients, we had 1 death, 1 aggravation, 1 improvement and 5 "cures" (62.5%). Based on data in the literature and our experience, we assessed the advantages of emergency surgery (immediate and definitive re-establishment of the carotid flow and vascularization of the hemisphere before installation of irreversible brain damage) and conditions suggesting chances of success: 1) diagnosis by noninvasive echo-Doppler of the cervical vessels and transcranial Dopler, without preoperative arteriography or CT-scan. 2) operation before 6 hours, 3) quality of the desobstruction, 4) no post-operative anti-coagulant treatment, 5) control of post-operative episodes of hypertension. PMID- 8713378 TI - [Atheromatous embolisms and cholesterol embolisms: medical treatment]. AB - In patients with an acute arterial occlusion, identification of the type of mechanism is important, because both prognosis and treatment differ for each type. The aorta is the most frequent source of arterial atheromatous emboli. Aortic arch plaques are therefore recognized as an independent risk factor for stroke, and plaques located on the thoracoabdominal aorta embolize in the visceral arteries or limb circulation. The treatment of risk factors seems the most effective preventive treatment. When atherosclerosis is patent, an anti platelet drug such as aspirin or ticlopidine is useful. When the embolus actually occurs, heparin avoids extension of thrombus and prevents its recurrence. Surgical treatment is logical but has not been supported by any randomized trial. Cholesterol cristal embolization evolves in 3 clinical forms: 1-the paucisymptomatic form, not diagnosed during subject's lifetime and only recognized in autopsy studies; 2-a benign form such as the blue toe syndrome or cutaneous livedo, with a spontaneous mild prognosis, and 3-a diffuse multisystemic form with a very poor prognosis. More than 80% of patients with the diffuse form die. When there is renal involvement, only 25% are still alive, with renal function after 6 months of follow up. Vascular surgery is limited to patients with aneurysms, which in themselves constitute a surgical indication. For all other patients, surgery is rarely indicated because 1-the source of cholesterol cristal embolization is not certain, 2-patients are usually too weak for a major surgical intervention, and 3-the necessary aortic clamping during surgery would induce a major risk of recurrence. Prevention is the most effective treatment because in 30% of patients, embolization is due to one of the following: anticoagulant drug, recent fibrinolysis, percutaneous angioplasty, vascular surgery, diagnosis angiography and/or coronarography. The medical treatment is mostly symptomatic: rest, warm conditions, appropriate dressing, antiplatelet drugs, hydration, and organ supply when necessary, principally to ensure renal function. In diffuse and multi-visceral embolization, either colchicine or corticosteroids adjuvant therapy might be useful Prostanoid drugs are also a possible adjuvant treatment. PMID- 8713379 TI - [Emboligenic aortopathies. Cholesterol embolisms: surgical treatment]. AB - Emboligenic aortopathies are defined as lesions of the aortic wall leading to the production of thromboatheromatous material which can migrate in fragments or entirely. Emboligenic aortopathy can occur in all parts of the aorta. Localized lesions are rare and usually involve the subrenal abdominal aorta or the isthma. Diffuse lesions involving several segments of the aorta are encountered more often. There are three anatomic types of lesions: ulcerated plaques, atheromatous ulcerations, an evrysmal disease of the aorta. Therapeutic indications depend on: 1) clinical presentation: fibrinocruoric emboli, distal microemboli, disseminated cholesterol emboli; 2) the unique or multiple nature of the emboli; 3) the anatomic lesion; 4) localization. PMID- 8713380 TI - [Protected carotid angioplasty and carotid stents]. AB - PURPOSE: Endovascular treatment of carotid stenoses. MATERIAL AND METHOD: 259 angioplasties of carotid stenosis are reported. The stenosis was an inflammatory stenosis or a restenosis post endarterectomy in 71 cases, atherosclerotic stenosis in 188 cases. Angioplasty was performed without cerebral protection in 123 cases including a group of 38 with atherosclerotic bifurcation stenosis. A cerebral protection (triple coaxial catheter) was used in 136 atherosclerotic bifurcation stenoses. RESULTS: There has been no complication related to the procedure in the 71 cases of nonatherosclerotic stenosis and the 14 cases of proximal carotid and siphon atherosclerotic stenosis. There has been 5% of dissection and 8% embolic complication in the group of 38 atherosclerotic bifurcation cases treated without cerebral protection. There has been no embolic complication in the 136 atherosclerotic bifurcation cases treated with cerebral protection. There has been 5% dissection in this group before stents and 0% after stents have been used in less then satisfactory results (61 stents placed). Restenosis has also decreased after stents (15% to 4%). CONCLUSION: Endovascular approach can now be considered for all types of carotid stenosis. A cerebral protection with temporary carotid occlusion is mandatory in atherosclerotic carotid bifurcation cases to rule out embolic complication. Complementary stents, when necessary, have reduced the immediate risk of dissection and of restenosis later on. PMID- 8713381 TI - [Endovascular treatment of arteries with cerebral destination: failures and limits]. AB - OBJECTIVE: Evaluate endovascular treatment of vessels irrigating the brain. Assess risks and indications of balloon angioplasty and stents. METHODS: Retrospective study in 38 patients (6 females, 32 males) who underwent revascularization from December 1990 to July 1995: 47 balloon angioplasties and 17 stents (36%). Three patients were asymtomatic, 17 had a past history of transient ischaemia, 5 had amauraosis, 9 signs of vertebrobasilar insufficiency and 2 had an ischaemia of the upper limbs. Endoluminal treatment was performed in 4 brachiocephalic trunks with implantation of 1 stent, in 7 common carotid arteries with 4 stents, in 24 internal carotid arteries with 9 stents, 2 osteal stenosis of the vertebral artery and one external carotid. RESULTS: There were no complications in patients treated for lesions of the brachiocephalic trunk, the subclavian artery and the vertebral arteries. Among the 7 patients with a stenosis of the common carotid artery, there was one death after reperfusion due to cerebral oedema. For the carotid internal, two groups of patients could be distinguished. In one group of 13 patients with restenosis of the internal carotid artery who were treated by balloon angioplasty, there were 3 transient episodes of ischaemia, one reversible hemiplegia and one silent infarction. A second group of 8 patients had atheromatous stenosis. One was treated by balloon angioplasty with one transient episode of ischaemia and the 7 others were treated with a stent without complications. The rate of neurological complications was 15.7% (6 deficits in 38 patients). The permeability after revascularization was verified at mid-term with repeated echo-Doppler examinations and by angiography one year after operation. Restenosis occurred early after one subclavian stent covered with a patch. Among the 16 Palmaz stents, one implanted in a post irradiation common carotid occluded after 2 months. The other 15 stents were patent at a mean follow-up of 18 months (2-56), i.e. 93%. There were 2 restenoses after balloon angioplasty in the group of carotid restenosis, i.e. 15%. CONCLUSIONS: Risk in balloon angioplasty of arteries irrigating the brain is a serious problem. Stenosis of the subclavian artery and the vertebral arteries appears to be a good indication. Lesions of the carotid bifurcation should not be treated with balloon angioplasty due to the risk of neurological complications. Among the restenosis after endarterectomy, only those lesions situated in the distal internal carotid are good indications. Stents have greatly improved treatment possibilities. They should be implanted whenever there is a risk of supra-aortic lesions and in certain lesions of the carotid bifurcation in high risk, patients. Their application in all situations cannot be proposed yet before long-term outcome is established. PMID- 8713382 TI - The role of angioplasty and stenting in the treatment of occlusive lesions of supra-aortic trunks. AB - PURPOSE: To review the authors' four-year experience with endoluminal treatment of stenotic and occlusive lesions of supra-aortic trunks, and to compare the results obtained with those achievable with more conventional surgical reconstruction. METHODS: The authors' four-year experience (7/1/91-6/30/95) with 30 endoluminal brachiocephalic procedures on 26 patients was reviewed retrospectively. The type of occlusive lesion encountered, arterial involvement, symptoms, and indications for treatment were noted. Patients were followed and reexamined at six-month intervals and information from this assessment constituted the source of information to determine success and patency rates. Only one patient was lost to follow-up at three months post intervention, at which time the recanalized subclavian artery was patent and the patient was asymptomatic. RESULTS: There were no operative mortalities or strokes. One patient developed a large cervical wound hematoma which required surgical evacuation. Immediate (technical) success was achieved in 27 arterial segments out of 30 which were approached with intention to treat, for a success rate of 90%. The three immediate failures involved totally occlusive lesions of the left proximal subclavian artery which proved unresponsive to retrograde transluminal recanalization. Long-term arterial patency was achieved in 24 of 30 instances, for a success rate of 80%. The three failure occurred respectively eight (right common carotid artery), 12 (right subclavian artery), and 18 months (left subclavian artery) after the initial procedure, and they all involved restenosis following angioplasty/stenting. CONCLUSIONS: The retrospective review reported herein demonstrates that angioplasty/stenting of focal stenotic and occlusive lesions of supra-aortic trunks would seem to produce immediate and long-term success rates which are quite acceptable, and may approach those achievable with more conventional surgical reconstruction. Further reporting of larger series of patients followed up for longer periods of time will be necessary for more definitive conclusions regarding these less invasive therapeutic options. PMID- 8713383 TI - [Antibiotic-impregnated prostheses: eclectic indications]. AB - OBJECTIVE: Infection is a major complication in vascular stents. Stents impregnated with gelatine and dipped in Rifampicin have been shown to resist methicillin-resistant Staphylococcus aureus in both animal experiments and in man. It has been suggested that all aorto-ilio-femoral stents should be treated. To evaluate this method, we reassessed all stent infections observed in our patients who had undergone revascularization of the lower limbs from January 1985 to 1994. We excluded stents implanted for ruptured aneurysms or implanted in patients with a past history of local infection on vascular stents. RESULTS: The rate of septic complications observed during the first year was 1% for all patients in the series, 0% for aorto-aortic and aorto-biiliac stents and 0.7% for aorto- bifemoral stents. These rates are similar to those reported in the multicentric study directed by Goeau Brissoniere using antibiotic impregnated stents. The extra cost involved in using such stents for aorto-ilio-femoral revascularization was estimated in this series at 2,180,000 Francs. The costs resulting from the three infections was estimated at 960,000 Francs. CONCLUSION: Based on the findings in this series, antibiotic impregnated stents should be indicated only in selected patients due to the extra cost: past history of local infection, ruptured aneurysms, femoro-tibial stents, cross or axillo-femoral revascularization for which the rate of stent infection is 6.3 - 3.2 and 1.4%, immunodeficient patients, multiple reoperations, post-irradiation arteritis and situations known to involve major risk of infection. PMID- 8713384 TI - [Antibiotic bonding on arterial prostheses: a necessity]. AB - The introduction of arterial grafts made of polyester impregnated with a protein network is an important advance. Besides preventing leakage, impregnation makes it possible to bind active molecules, particularly antibiotics. Rifampicin is one of the antibiotics with an activity spectrum adapted to micro-organisms involved in graft infections and with a particular affinity for the protein network impregnating certain grafts. Rifampicin's affinity for gelatine means that large amounts of the antibiotic can be bound simply by dipping the graft in an antibiotic solution. This technique has been shown to be useful for both prevention and treatment in an experimental model. These preliminary findings have led to a multicentric randomized European trial which has provided encouraging results showing a decrease in the overall and early infection rates. PMID- 8713385 TI - [Treatment of an exposed femorol-popliteal bypass: ex-situ replacement]. AB - From December 1990 to July 1995 we performed 171 sub-inguinal revascularizations including 35 popliteal revascularizations and 146 revascularizations of an artery in the leg or foot. Five cases of infection were observed within a delay of 7 and 25 days after the operation. There were 3 men and 2 women (mean age 78 years). Four femoro-tibial bypasses were made for critical ischaemia (2 necroses of the toes, one eschar of the heal, one stage III). There was one femoro-popliteal bypass which was associated with a femoro-femoral for necrosis of the toes. Two bypasses were made with polytetrafluoroethylene, one with Dacron and two with the greater saphenous vein. Signs of sepsis were bleeding in 2 patients who had a venous bypass and septicaemia in 2 patients. Local skin necrosis and/or apparently infected discharge or patent pus were seen in all patients. Staphylococcus aureus was found in 4 patients and Enterobacter cloacae in one. Revascularization was done with an extra-anatomic bypass in 4 patients and with a cryopreserved in situ allograft in 1. Mortality was 20% and amputation rate was 40%. All exposed bypasses were infected but the severity of the infection varied depending on the causal germ, general signs and ischaemia of the limb. Conservative treatment has its limits: 1) intact anastomoses, 2) absence of bleeding, 3) patent bypass, 4) absence of generalized sepsis. Results of in situ revascularization depend on the virulence of the causal germ. Radical treatment (explanation + extra-anatomic revascularization) still has indications in infected infra-inguinal bypass surgery. PMID- 8713386 TI - [Exposed infra-inguinal bypass: conservative treatment or in situ replacement]. AB - Infection or exposure of a infra-inguinal bypass is a severe complication which can lead to death or amputation. With conservative treatments, the bypass is left in situ or replaced with an autologous vein or an allograft. Results in a series of 393 consecutive conservative treatments for infra-inguinal revascularizations were analyzed retrospectively. Sixteen patients were studied and divided into 2 groups: Group 1 included 8 patients who had no bleeding or signs of infection and who were treated locally. Group II included 8 patients with bleeding or signs of infection in whom the bypass was replaced by an in situ autologous vein (n = 4) or an arterial allograft (n = 4). Mortality was 12.5% and the rate of amputation was 12.5%. These percentages are in agreement with reports from other authors who also propose this therapeutic option. Local treatment should be reserved for infection free of complications due to thrombosis, hemorrhage or signs of a septic syndrome. Other cases should benefit from in situ replacement with a vein or an arterial allograft. In some unsuccessful cases there are a few indications for extra-anatomic prosthetic bypasses. PMID- 8713387 TI - [Treatment of prostheto-digestive fistulas using in situ prosthetic bypass]. AB - Secondary aorto-enteric fistula is one of the most serious complications of abdominal aortic reconstruction. Conventional management includes removal of all infected prosthetic graft, oversewing of aortic stump and restoration of lower limbs blood flow by extraanatomic bypass grafting, reporting high rates of mortality, limb loss, and even infection of the extraanatomic grafts. Dissatisfied by these results, frequently, due to aortic stump blowout or extraanatomic by-pass reinfection, some authors attempted a more conservative approach with au in situ replacement by a new synthetic graft. The aim of this paper was to verify the role of in situ graft replacement. From December 1989, 8 patients with secondary aorto-enteric fistula underwent in situ PTFE graft replacement. One patient (12.5%) died perioperatively for acute myocardial infarction. No limb loss occurred. One patient died after 44 months from pulmonary neoplasia without signs of graft infection. The others are doing well at 34 months follow-up. The authors suggest that, in selected patients, in situ prosthetic graft replacement provides better early and late results than extranatomic bypass. PMID- 8713388 TI - [Treatment of prostheto-digestive using arterial allograft]. AB - From October 1988 to March 1995, we operated 22 patients for fistulization between the prosthesis and the digestive tract to remove the in situ allograft. The delay between the initial operation and treatment for fistulization was 7.3 +/- 4 years. In these patients who had undergone multiple operations (2.5 +/- 1.9 operations per patient), the infected prosthesis was made of Dacron in 21 cases and polytetrafluoroethylene in one. The procedure was planned beforehand in 21 cases who benefited from a complete preoperative work-up and was required in an emergency situation in 6 for digestive bleeding (5 cases) or an abscess of the Scarpa (1 case). Among the patients with an emergency operation, three of the procedures were conducted within a single operative time and three with two separate procedures. The allografts were aorto-aortic tubes (n = 3), aortobifemoral bypasses (n = 14), aorto-iliac bypasses (n = 4) and one aorto femoral-iliac bypass. Organ revascularization was associated in 8 patients. Seven patients (32%) died post-operatively. Five of them had undergone an emergency procedure. An amputation was required in 2 patients, one at the time the allograft was implanted and the second due to ischaemia despite a permeable allograft. None of the patients had to be amputated due to failure of the allograft. Mean follow-up was 36.6 +/- 20 months. There were 4 deaths post operatively due to digestive bleeding in 2. The aortic allograft was dilated in 4 patients without re-operation. Thrombosis of the allograft branch occurred in 4 patients, including 3 who had been re-operated successfully. Despite these still perfectable results, treatment of secondary digestive-prosthesis fistulae with an in situ allograft constitutes a real progress in terms of patient survival and preservation of the limb in high-risk patients. PMID- 8713389 TI - [Forefoot gangrene and infra-crural bypass: simultaneous amputation]. AB - Patients presented for amputation mostly have chronic limb ischaemia caused by atherosclerosis, with signs of severe arterial insufficiency including rest pain, non-healing skin lesions, ulceration or gangrene. Foot infections, especially in diabetic patients, are often multimicrobial, deeply invasive and frequently require aggressive measures, like debridement and drainage or partial open forefoot amputation in addition to broad-spectrum antibiotics, in patients with critical limb ischaemia and limited necrosis and forefoot gangrene, distal bypass surgery is the treatment of choice. The main question is whether amputation should be performed simultaneously or in a secondary stage. Our own experience deals with 342 femorocrural and femoropedal bypass grafts for the treatment of critical limb ischaemia. The results showed no significant difference in graft patency between crural and pedal grafts. Clinical factors like diabetes mellitus, poor distal run-off and site of the distal anastomosis had no adverse effect on the functioning and patency of the graft. In this series we found that in diabetic patients significantly more amputations were required because of persistent foot infection. Since in these patients amputation was performed in a secondary stage, we changed our policy to simultaneous amputation. After completion of the bypass, closure and coverage of all the wounds, the gangrenous part is amputated. In case of deep, wet or infectious gangrene of the forefoot, an open transmetatarsal amputation is performed. Using this approach we have further increased limb-salvage and especially the number of usuable limbs. PMID- 8713390 TI - [Forefoot gangrene and infra-crural bypass: sequential amputation]. AB - When revascularizing for gangrene, deciding on amputation depends on the risk of infection due to trophic injury both for the foot and for the bypass and on the requirement to attempt amputation at the correct level at the first operation. In our experience, the importance of infection has led us to propose the following sequence: if the lesions are infected--immediate amputation, differed revascularization after the infection has been controlled; if the lesions are dry early vascularization followed by amputation either during the same operation with strict separation of the two operative fields or a few days later, particularly if the level of the amputation has been determined. PMID- 8713391 TI - Interposition vein cuffs. AB - A vein cuff interposed at the distal anastomosis between a prosthetic vascular graft and a recipient infrageniculate artery improves the chances of continued patency of the graft, at least in the short and medium term. The mode of effect appears to be suppression or modification of anastomotic myointimal hyperplasia (MIH). In the event of graft failure the recipient artery and run-off vessels remain free from MIH and their patency is preserved thereby improving the prospects for further vascular reconstruction and limb salvage. The mechanisms by which interposition vein cuffs might modulate MIH are reviewed. Experimental evidence is described to show that the geometry of a cuffed anastomosis promotes a characteristic haemodynamic flow structure with a stable vortex. It is suggested that this vortex exerts greater shear stress upon the wall of the artery than the normal laminar flow observed in conventional anastomoses. High shear stress is known to inhibit MIH. PMID- 8713392 TI - [Importance of proper implementation of the case history in the prevention of stomatological diseases]. AB - In connection with ageing of population and an increasing number of older patients visiting the dental practice with multiple diseases, special prevention of possible medical complication during dental treatment is necessary. The advances in dentistry increase the chance of these complications. There is however no consensus about history taking in dental practice. For getting the maximum amount of information about the patient's health a Risk-Related Medical History (RRMH) patient-administered is proposed, that is checked by the dentist. The RRMH was composed in accordance with the modified ASA physical risk score. It allows to dyfine general patient's physical condition and to advise measures for prophylaxis of medical complication. The medical questionnaire showed a high sensitivity and specificity. In a study among 5000 patients the results of this history was analysed. The RRMH is simple and comfortable, it allows to obtain relible medical information in short time both for the dentist and for the patient. PMID- 8713393 TI - [Certification of oral hygiene agents from a microbiological viewpoint]. AB - The authors validate the necessity of hygienic certification including microbiological control of perfumes and cosmetics, to which oral hygiene means belong. They consider that the absence of sanitary regulations for some goods, toothpastes included, is one of the causes, responsible for the absence of a proper sanitary control on the part of State Epidemiological Survey of the Russian Federation. In accordance with the resolution "On Certification of Perfumes and Cosmetics" issued in 1994, Central Research Institute of Stomatology is appointed the central institution responsible for certification of means of oral hygiene, and at present its main task is development of sanitary regulations, standards, methods of investigation, and training of personnel for institutions which are to carry out certification. PMID- 8713394 TI - [The effect of pinocadinene on the clinico-immunological status of patients in the combined treatment of periodontitis]. AB - Local immunity parameters of patients with periodontitis were studied before and after pinocadinen therapy. Local application of pinocadinen had a favorable impact on local immunity and protective properties of the saliva. The drug promoted a rapid arrest of the inflammatory process in the periodontium. PMID- 8713395 TI - [Laser physiotherapy of stomatological diseases]. AB - Modern methods of laser prophylaxis and physiotherapy of numerous dental diseases in cariesology, endodontia, periodontology, surgery, orthodonty, pedodontics, implantology, etc., using new-generation laser technologies, are reviewed. PMID- 8713396 TI - [Bacterial activity in the products of laser destruction of human dental enamel and dentin]. AB - Viability of Streptococci, Staphylococci, and Bacillus subtilis in products of destruction of hard dental tissues of man by submillisecond pulses emmitted by erbium lasers YAG:Er and YSGG:Cr,Er was investigated. Bacterial viability was found to depend on the energy density and duration of laser exposure. Possible mechanisms of bacterial destruction by pulses of submillisecond duration emmitted by erbium laser are discussed. PMID- 8713397 TI - [Analgesic action of ketorol in clinical maxillofacial surgery]. PMID- 8713398 TI - [Asymmetry of the human facial nerve]. AB - Measurements of the right and left facial skull were carried out in 6 different series of skulls: two europeoid, Russian and Georgian; two mongoloid, Buryat and Eskimo; and two skulls of mixed origin, Khakass and Udmurt ones, in order to assess the bilateral asymmetry. The measurements included 18 signs characterizing the width, height, and length of the facial skull. Sums of all right-side and left-side signs were the integral parameters of the right and left halves of the facial skull. A compensatory type of asymmetry variations was revealed. All the components of the facial skull regarded individually demonstrate clear asymmetry, whereas the total summary size of both halves of the facial skull is similar. The cause of the symmetrical combination of whole parts in the presence of evident asymmetry of various elements of the facial skull is differently directed asymmetry of various extent. The results are discussed in the context of the evolution of human functions and the relevant transformation of the morphologic structure in the anthropogenesis. PMID- 8713399 TI - [A method for the surgical treatment of maxillary micro- and retrognathism]. AB - The proposed method of surgical treatment of maxillary micro- and retrognathia radically differs from the known methods: osteotomy of the osseous base of the hard palate is carried out parallel to the vomer to reach the level of the first premolars by a median dissection of the mucosa and periosteum, and vertical osteotomy of the anterior portion of the vomer is carried out. Horizontal osteotomy is performed from the vestibule of the mouth. Subsequent osteotomy of the anterior and lateral surfaces of the upper jaws under mucosal periosteal "tunnels" and detachment of the maxillary tuberosities from the coracoid processes permit a full-scale displacement of the bone fragment forward to attain the position planned for it. The method was clinically used in 7 patients with good functional and esthetic results. PMID- 8713400 TI - [Removal of the sagging of the tissues in the submental area]. AB - Twenty out of 100 patients operated on to remove skin excess on the face and neck had slackening tissue under the chin. Based on published data and their findings, the authors developed operations to remove slackening tissue under the chin and carried out such operations in 64 patients. Isolated lipectomy failed to bring about good results in cases when there were indications for removal of skin excess on the face neck (10 patients). Simultaneous removal of skin excess on the face and lipectomy helped attain good results in 50 patients. The operation is possible in patients with a low position of the sublingual bone, although the results are not so good. Lipectomy in the area under the chin is recommended as a method supplementing operation for removal of skin excess on the face and neck. PMID- 8713401 TI - [Reconstructive operations on the head and neck using a microsurgical technic]. AB - Eighty-one patients with defects and deformations of the head and neck of various origin were examined and treated. Microsurgical autotransplantation of vascularized tissue complexes was carried out in all the cases. Profound preliminary examinations of patients helped objectively define the indications for the use of this method. Special attention was paid to the choice of donor zone. Due to correct selection of a complex graft, good functional and cosmetic results were attained. Despite repeated corrective operations in 16 patients, virtually complete social and communal rehabilitation was attained in 74 (91.4%) patients by using a limited number of donor areas. The authors persuasively demonstrate the advantages of microsurgery in repair of complex and extensive defects on the head and neck. PMID- 8713402 TI - [The polymerization process of Ethacryl acrylic plastic studied by infrared spectroscopy]. AB - The kinetics of hardening of Ethacryl plastic used in making the base of removable dentures was studied by infrared spectroscopy. The authors demonstrate the potentialities of this method for a reliable control of the residual monomer influencing the physicomechanical and hygienic properties of dentures in the course of wearing. PMID- 8713403 TI - [Nonspecific resistance factors of the body in middle-aged and elderly patients with denture stomatitis]. AB - Various manifestations of intestinal dysbacteriosis, from latent to diffuse ones involving the entire gastrointestinal tract, were detected in elderly and senile patients wearing removable dentures; in 36% of them nonspecific bacteremia was detected. These manifestations were most expressed in patients with allergic stomatitis. Patients wearing removable dentures and suffering from prosthesis induced stomatitis should be followed up by gastroenterologist. PMID- 8713404 TI - [Methods for estimating and predicting the demand for stomatologic personnel]. AB - Analysis of the WHO Global Data Bank and Monitoring of Dental Diseases Prevention (tendencies of dental caries incidence in the world and in all regions of the former USSR) helped the author derive a formula for estimating the need in dentists. The number of dentists needed may be predicted for up to 15 years by assessing the tendencies in caries intensity in the population of a region. PMID- 8713405 TI - [The characteristics of the anti-inflammatory action of preparations used in dentistry]. PMID- 8713406 TI - [Hydroxyapol and Colapol in dental practice]. PMID- 8713407 TI - [A contact-free method for computerized functional assessment of mandibular movements]. AB - A specialized computer system for real-time study of some dynamic characteristics of the maxillodental system has been designed. The system consists of a TV unit compatible to IBM PC/AT and software. The method of non cintact recording of dynamic characteristics of the mandible permits, among other things, measurements and comparison of mandibular movements before and after orthodontic treatment and, hence, may be used for assessing the results of occlusion correction. PMID- 8713408 TI - [Autonomic regulation in periodontitis]. AB - Presents analysis of the status of different levels of autonomic innervation in patients with generalized periodontitis. Sympathetic activation at both suprasegmentary and segmentary levels revealed in all the patients is a pathogenetic component in the disease development. A stable correlation has been detected between the degree of segmentary sympathetic activation and severity of generalized periodontitis. PMID- 8713409 TI - [A rare case of a preauricular fistula]. AB - A female patient with a preotic fistula caused by a defect in the development of the first branchial arch is described. Repeated surgeries for lateral fistula of the neck brought no success. The pathogenesis of the defect and difficulties in its diagnosis are discussed; the authors claim that fistulography with contrast agents may be effective. Histologic verification is needed in all the cases. PMID- 8713410 TI - [Delivery of care to patients with facial wounds in Russia and the Soviet Union in the 20th century]. PMID- 8713411 TI - [The endogenous intoxication syndrome in sepsis]. PMID- 8713412 TI - [Assessment of the severity of patient status in sepsis and septic shock]. AB - A score for quantitative evaluation of the severity of clinical status has been created for a more objective assessment of the clinical status of patients with sepsis and septic shock. The system includes five sections: physiological parameters, level of respiratory support, age, localization of the focus, concomitant diseases. The final score represents a sum of scores for the above sections. ROC analysis demonstrated a higher prognostic value of the proposed score in comparison with the APACHE-II for patients with grave sepsis. PMID- 8713413 TI - [The use of perftoran in clinical medicine]. PMID- 8713414 TI - [Aspects of perfluoro-organic hemo-protection]. AB - As effective gas-transporting substances, organoperfluorine compounds are characterized by hemoprotective properties. Pretreatment with fluorocarbon helps level the direct unfavorable effect of oxygen on the blood upon its contact oxygenation, which was confirmed by the results of studying red cell structure and function. PMID- 8713415 TI - [Changes in hemodynamics and the sympathetic-adrenal system during combined transcutaneous electric nerve stimulation in minor surgical interventions]. PMID- 8713417 TI - [Epidural electric nerve stimulation]. PMID- 8713416 TI - [The interrelationship of changes in carbohydrate-energy metabolism and potassium homeostasis in children with diffuse suppurative peritonitis]. AB - The parameters of potassium homeostasis (potassium concentration in the plasma, red cells, and urine and potassium to sodium ratio in the urine) and of carbohydrate-energy metabolism (CEM) assessed from the activities of the key enzymes of glycolysis and pentose phosphate route of glucose oxidation were determined in 52 patients on day 2 postoperation. Relationships between potassium metabolism and CEM status were revealed in children with diffuse peritonitis. Disorders in potassium homeostasis were associated with CEM changes. In children with a favorable course of the disease the adaptation changes in the metabolic system, characterized by increased intensity of glucose oxidation and intactness of cellular energy homeostasis, permit the maintenance of potassium homeostasis at optimal potassemia of low therapeutic value. In case of a lethal outcome failure of therapeutic measures aimed at correction of potassium homeostasis is coupled with CEM disorders manifesting by reduced glucose oxidation after the pentose phosphate route, reduced concentration of adenyl nucleotides, disordered membrane permeability, and accumulation of organic acids. PMID- 8713418 TI - [High-resolution ECG in the diagnosis of cardiac electrical instability and the prognosis of sudden death]. PMID- 8713419 TI - [Catheterization of the superior bulb of the internal jugular vein and its identification based on blood gas analysis in heart surgery patients]. AB - Methods of catheterization of the upper internal jugular bulb are described. The authors recommend to identify it basing on the difference between gaseous composition of blood samples collected simultaneously at two levels. The informative value of catheterization of the upper internal jugular bulb for assessment of brain metabolism is demonstrated. PMID- 8713421 TI - [Indirect electrochemical detoxication]. PMID- 8713420 TI - [Intensive therapy in diabetic ketoacidotic coma]. PMID- 8713422 TI - [A program of combined intensive therapy in critical ischemia of the lower extremities]. AB - Ultraviolet irradiation (UVI) of the blood, adsorption methods of detoxication, hyperbaric oxygenation (HBO), and antioxidant therapy are pathogenetic and etiotropic methods of treatment in multiple-modality conservative therapy of patients with ischemia of the lower limbs. The EKROMI UVI method is rapid and no less effective then the traditional exposure of the blood after Knott. Enterosorption (ES) may replace hemosorption (HS) in lower limb ischemia with good effect; noteworthy that it is a safer method of detoxication. Addition of antihypoxants and antioxidants (AO) to therapeutic complexes administered before HBO improved the efficacy of this procedure. Use of the UVI-HS(ES)-AO-HBO-HS(ES) UVI scheme for intensive care for obliterating diseases of the arteries levels the shortcomings of each of the methods of the complex and potentiates the efficacy of each of them. PMID- 8713423 TI - [The correction of post-poisoning damages to the myocardium caused by carbophos]. AB - Lipid peroxidation processes and levels of oxidative metabolism enzymes succinate and lactate dehydrogenases, as well as morphometric parameters of cardiomyocytes were studied in rats after exposure to malathion. The efficacy of oxymethacil and atropin correction of postintoxication injuries to the myocardium was assessed. The new drug oxymethacil is recommended to be included in the treatment protocols during the first 48 h after grave poisoning with malathion. PMID- 8713424 TI - [Severe neurogenic pulmonary edema as a result of rupture of a giant intracranial arterial aneurysm]. PMID- 8713425 TI - [Legal problems in the activities of the anesthesia-resuscitation specialist]. PMID- 8713426 TI - [Termination of long-term artificial pulmonary ventilation ("weaning from an APV apparatus"). Functional criteria and methodological principles]. PMID- 8713427 TI - [Gastrointestinal tract paralysis in surgery and methods for its treatment]. PMID- 8713428 TI - [Magnesium and its balance]. PMID- 8713429 TI - [Activity of threonine deaminase and biosynthesis of avermectins in the culture of Streptomyces avermitilis]. AB - The influence of ammonium, threonine, isoleucine and valine on the activity of threonine deaminase and the biosynthesis of avermectins in the culture of two mutants of Streptomyces avermitilis, i.e. a sensitive one and a resistant one with respect to alpha-amino-beta-oxyvaleric acid, a threonine antimetabolite, was studied. It was shown that the synthesis of threonine deaminase was induced by threonine and valine in the mycelium of both the mutants. The level of threonine deaminase was higher in the mycelium of the antimetabolite resistant mutant. The antibiotic activity of the resistant mutant was lower while the relative content of the group B avermectins in the pool of the synthesized avermectins was higher than that in the culture of the sensitive mutant. PMID- 8713430 TI - [Reaction of acylation of the antineoplastic antibiotic bleomycetin]. AB - The reaction of benzoylation of bleomycetin, a glycopeptide antibiotic, and its cuprum complex was studied with the chloranhydride and carbodiimide methods. Acylation of the antibiotic cuprum complex by an equivalent quantity of benzoyl chloride resulted in formation of a mixture of mono- and disubstituted derivatives while with the use of a 3-fold excess of chloranhydride it was possible to obtain dibenzoylbleomycetin as a sole reaction product. Interaction of the bleomycetin cuprum complex with activated benzoic ether resulted in formation of a product monosubstituted by the antibiotic spermidine fragment. After acylation of the free antibiotic by the carbodiimide and chloranhydride methods tri- and tetrabenzoyl derivatives of bleomycetin were isolated respectively. PMID- 8713431 TI - [Action of lovastatin--an inhibitor of cholesterol biosynthesis on bacterial bioluminescence]. AB - The action of lovastatin, a competing inhibitor of 3-hydroxy-3-methylglutaryl CoA reductase, on bacterial bioluminescence was studied. The lovastatin lactone form and sodium salt of mevinolinic acid inhibited bacterial luciferase in vitro but did not affect bioluminescence of the intact cells of the luminous bacteria. The inhibition was found to be of a competing character in regard to aliphatic aldehyde, the bacterial luciferase substrate. Conditions under which the bioluminescence inhibition was proportional to the lovastatin concentration in the incubation mixture and a bioluminescence method for quantitative determination of the inhibitor were developed. PMID- 8713432 TI - [Cell cultured of Macleaya cordata--a producer of antimicrobial benzo[c]phenanthridine alkaloids]. AB - It was shown that a suspension culture of Macleaya cordata accumulated benzo(c)phenanthridine alkaloids in the cell biomass. By the composition of the alkaloids and their content the cell culture markedly differed from the intact plant and callus tissue. The benzo(c)phenanthridine compounds in the cell culture were mainly represented by the reduced forms: dihydrosanquinarin and dihydrochelirubin. In the green plant they were mainly represented by the oxidized forms: sanquinarin and chelerithrin. The cell culture practically contained no chelerithrin characteristic of the intact plant. PMID- 8713433 TI - [Antibiotic sensitivity of hospital strains of Staphylococcus]. AB - Seven hundred and six clinical strains of Staphylococcus spp. isolated in different cities of the former USSR in 1986-1992 were investigated. It was shown that the isolates had multiple drug resistance. The hospital strains of Staphylococcus aureus more frequently contained the plasmids with the molecular weight of 21.0 and 18.0 MD as well as the plasmids with the molecular weight of 1.4 MD determining erythromycin resistance. The plasmids with the molecular weight of 3.0 and 2.6 MD were detected which determined streptomycin and tetracycline resistance respectively. Lincomycin and carbenicillin resistance was determined by the chromosomal genes. The elimination studies demonstrated that gentamicin resistance could be determined by the extrachromosomal genes. PMID- 8713434 TI - [Biological characteristics and antibiotic sensitivity of Acinetobacter strains]. AB - Fifty Acinetobacter strains were investigated. Their differentiation according to the species position showed that 30 strains belonged to A. calcoaceticus and 20 strains belonged to A. lwoffi. It was found that the Acinetobacter strains had multiple drug resistance. They showed 100 per cent resistance to ampicillin, carbenicillin, erythromycin, methicillin, oxacillin, tetracycline, oleandomycin, streptomycin, rondomycin and lincomycin. According to the results of the study the most efficient antibiotics were cefazolin, cefotaxime, ceftazidime and polymyxin. The Acinetobacter hospital strains contained conjugative plasmids of different incompatibility groups. 50 per cent of the Acinetobacter strains contained conjugative R plasmids determining the resistance to streptomycin, kanamycin and tetracycline and referring to the T, J, M and N incompatibility groups. Their molecular weights were 42.0, 35.0, 30.0 and 50.0 MD respectively. PMID- 8713435 TI - [Survival of Escherichia coli in human blood serum as a function of R-plasmid content]. AB - Thirty four strains of Escherichia coli of the pathogenic and nonpathogenic serogroups were tested for their survival in normal human blood serum as dependent on the content of the R plasmids. It was shown that the capacity for the survival of the E. coli strains of different origin was widely distributed. The value of the survival associated with the content of the R plasmids was higher in the E. coli strains of the pathogenic serogroup by comparison to that in the ordinary cultures. The capacity for the survival of the E. coli strains in the serum was more frequently accompanied by the tetracycline resistance. PMID- 8713436 TI - [Evaluation of the effectiveness of the use of pefloxacin in combined radiation thermal injury]. AB - The efficacy of pefloxacin, a novel promising antibacterial drug of the group of fluoroquinolones, was studied on mice against doxycycline in the prevention and treatment of infectious complications of radiation/thermal injury. According to the data on the effect of the antibiotics on the resistance to exogenic infection and the development of dysbacteriosis pefloxacin was recommended for the use in the scheme of combined therapy of radiation/thermal injury. PMID- 8713437 TI - [Microflora of purulent necrotic leisions of the foot in patients with diabetes mellitus]. AB - Microflora of purulent necrotic lesions of the foot in 14 patients with diabetes mellitus was investigated by aerobic and anaerobic procedures. It was shown that the process was due to the same species of pathogenic microorganisms irrespective of the lesion severity. However, the number of the pathogens in the associations and the dissemination level per a patient were higher when the lesions were extended. The tissue microbial dissemination in the purulent foci was also higher in the patients with wounds complicated by marked inflammation of the surrounding tissues and the tissues under the lesion. The time course of the changes in the foot microflora during the target-aimed antibacterial therapy was followed up. PMID- 8713438 TI - [Effectiveness of pefloxacin in the surgical clinic]. AB - Pefloxacin (Rhone-Poulenc, France) was used prophylactically and therapeutically in 8 and 21 surgical inpatients respectively. The majority of the Enterobacteriaceae isolates were susceptible to pefloxacin (91 to 100 per cent). The isolates of Acinetobacter spp. and Pseudomonas aeruginosa were less susceptible (77 and 33.3 per cent respectively). The isolates of Staphylococcus aureus and S. epidermidis were mainly susceptible to the drug. The high antimicrobial activity of pefloxacin used therapeutically well correlated with the favourable time course of the main disease. When pefloxacin was used prophylactically the results were positive in all the cases. The drug tolerance in the majority of the cases was good. Only 2 patients developed allergic reactions as multiple minipunctate rash. After discontinuation of the drug use and short-term course of the desensitization the clinical sings of the allergic reactions disappeared in 3 days. PMID- 8713439 TI - [Use of unified methods in microbiologic monitoring of penicillin resistance in Pneumococcus]. AB - The microbiological surveillance of pneumococci according to the Protocols for the International Surveillance of Pneumococci Resistant to Penicillin and Other Antimicrobial Agents developed by Prof. M. Jacobs (USA) and Prof. P. Appelbaum (USA) made it possible for the first time in Russia to detect the strains with moderate resistance to penicillin. It is necessary to revise the current Russian recommendations for the laboratory diagnosis of pneumococcal infection and the determination of antibiotic susceptibility so as to participate in the united programme of the International Surveillance of Penicillin Resistance in Streptococcus pneumoniae. PMID- 8713440 TI - [Effect of fluoroquinolones on the interaction of phagocytic cells with pathogens, lymphocyte function and immune response]. PMID- 8713441 TI - [Breast feeding and the nutritional status of Chilean children]. AB - The purpose of this work was to describe the diet of children under 18 months of age and its relationship to nutritional status. For this purpose, in 1993 a prevalence study was carried out on children who received care from the National Health Services System of Chile. The Chilean public health system provides care to 75% of children under 6 years of age and especially to children of families in the lower strata of income distribution in the country. Participants in the study were 9330 children under 18 months old who were randomly selected from 102 of the 320 urban clinics throughout the country. The type of food these infants had received the day prior to the interview (breast milk exclusively, breast milk plus solid food, formula exclusively, formula plus solid food) and their nutritional status relative to the standards of the National Center for Health Statistics (United States of America) and of WHO were determined. Children with z values for weight-for-age between -1.0 and -2.0 standard deviations were considered at risk for malnutrition, and those with z values under 2 standard deviations were classified as malnourished. The prevalences of exclusive breast feeding at the first, third, and sixth months of life were 86.5%, 66.7%, and 25.3%, respectively. Some 12.1% of the participants showed low weight for age; 30.7%, low height for age; and 35.7%, overweight. The magnitude of weight-for-age deficiency was 1.2 to 5 times greater among children who were fed milk substitutes than among those who received breast milk. Breast-feeding also had a positive effect on height. These results confirm the benefits of exclusive breast feeding until 6 months of age, the need to supplement the child's diet with solid food after that age, and the breast-feeding's protective effect on the nutritional status of children of all the ages studied. PMID- 8713442 TI - [Evaluation of a training program for traditional birth attendants in Quetzaltenango, Guatemala]. AB - In 1992, a training program for traditional birth attendants (TBAs) was initiated in Quetzaltenango, Guatemala, in order to improve the identification and treatment of neonatal disorders and high-risk pregnancies. This program was evaluated by comparing the pattern of hospital referrals by TBAs before and after the training. The study design consisted of a community intervention evaluated by means of several cross-sectional surveys. Comparisons were made between a group of TBAs who had received the training (intervention group) and another group of TBAs from nearby communities who had not received the training (control group). The outcome variables included the TBAs' ability to correctly identify obstetric complications, the point in time at which they sent mothers to the hospital, and perinatal mortality among the children whose mothers were referred to the hospital. Of the 854 women eligible, 845 participated in the study. Following the training program, there was a 200% increase in the number of mothers referred to the hospital by the TBAs. There was also improvement in the ability of the TBAs in the intervention group to refer women with obstetric complications to the hospital, although the effect of the program appeared to have been minimal on both that ability and the reduction of perinatal mortality. Nevertheless, the latter effect should be more precisely determined, since a decrease in perinatal mortality was observed in both the intervention and control groups, with no statistically significant difference. PMID- 8713443 TI - [Influence of PAHO publications on the scientific health production in Latin America and the Caribbean]. PMID- 8713444 TI - [The 30th Meeting of the Advisory Committee on Health Research]. PMID- 8713445 TI - C-fos expression during vocal mobbing in the new world monkey Saguinus fuscicollis. AB - In order to find brain areas involved in the vocal expression of emotion, we compared c-fos expression in three groups of saddle-back tamarins (Saguinus fuscicollis). One group, consisting of three animals, was made to utter more than 800 mobbing calls by electrical stimulation of the periaqueductal grey of the midbrain (PAG). A second group, consisting of two animals, was stimulated in the PAG with the same intensity and for the same duration as the first group but at sites that did not produce vocalization. These sites lay somewhat medial to the vocalization-eliciting sites. A third group, consisting of two animals, was stimulated at vocalization-eliciting sites in the PAG but with an intensity below vocalization threshold. Fos-like immunoreactivity that was found in the vocalizing but not in the non-vocalizing animals was located in the dorsomedial and ventrolateral prefrontal cortex, anterior cingulate cortex, ventrolateral premotor cortex, sensorimotor face cortex, insula, inferior parietal cortex, superior temporal cortex, claustrum, entorhinal and parahippocampal cortex, basal amygdaloid nucleus, anterior and dorsomedial hypothalamus, nucleus reuniens, lateral habenula, Edinger-Westphal nucleus, ventral and dorsolateral midbrain tegmentum, nucleus cuneiformis, sagulum, pedunculopontine and laterodorsal tegmental nuclei, ventral raphe, periambigual reticular formation and solitary tract nucleus. For some of these structures (e.g. anterior cingulate cortex and periambigual reticular formation), there is evidence also from electrical stimulation, lesioning and single-unit recording studies that they are involved in vocal control. For other structures (e.g. lateral habenula, Edinger-Westphal nucleus), the available evidence speaks against such a role. Fos activation in these cases is probably related to non-vocal reactions accompanying the electrically elicited vocalizations. A third group of structures consists of areas for which a role in vocal control cannot be excluded but for which the present study presents the first evidence for such a role (e.g. claustrum and sagulum). These structures deserve further studies using more specific methods. PMID- 8713446 TI - Accurate synapse regeneration despite ablation of the distal axon segment. AB - In each body ganglion of the leech Hirudo medicinalis there is a single S-cell. After an S-cell axon is severed, it regenerates along its surviving distal segment and reconnects with its synaptic target, the axon of the neighbouring S cell. In approximately half the cases the regenerating axon forms a temporary electrical synapse specifically with the distal segment, which remains active and connected to the target, thereby functioning as a splice until regeneration is complete. To determine whether the distal axon segment is required for successful regeneration, distal segments of severed S-cell axons were ablated by intracellular injection of bacterial protease. Fifty-seven preparations were examined from 2 to 212 days after injection of the axon segment. The extent of S cell axon regeneration was assessed electrophysiologically by intracellular and extracellular recording, and anatomically by intracellular injection of markers followed by light microscopy and electron microscopy. The S-cell axons regenerated successfully in almost 90% of animals examined after 2 weeks or more. In a further four animals the target S-cell was ablated in addition to the distal axon segment, permanently disrupting conduction along the S-cell pathway. Nevertheless, the regenerating axon grew along its usual pathway and there was no evidence that alternative connections were formed. It is concluded that, although the distal axon segment can provide a means for rapid functional repair, the segment is not required for reliable regeneration of the axon along its usual pathway and accurate formation of an electrical synapse. PMID- 8713447 TI - Olfactory bulb repetitive stimulations reveal non-homogeneous distribution of the inhibitory processes in the rat piriform cortex. AB - Optical signals were recorded in the in vivo rat piriform cortex in response to a burst of seven electrical stimulations (100 ms interval) delivered in the olfactory bulb. Based on the recorded responses, three types of signal could be identified according to the relative amplitude of their monosynaptic and disynaptic components. The dysynaptic component had a larger (type 1) or an equal amplitude (type 2) compared with the monosynaptic one. Type 3 exhibited only the monosynaptic component. Type 1 represented 96% of the first response. The second response was characterized by an increase in type 3 signals (39%). The remaining type 1 signals were lower in amplitude when compared with the first response. The responses to the last five stimulations did not differ from one another but were different from the first two (type 1, 74%; type 2, 7.8%; type 3, 18.2% on average). The spatial distribution of these three types of signal was analysed by dividing the piriform cortex into several areas. These areas were not homogeneous in the percentage of each signal type: the percentage of type 3 signals was highest (approximately 30%) in the area near the lateral olfactory tract and < 10% in the most posterodorsal area. Thus the level of inhibition remained high in some piriform areas whereas it decreased rapidly in others, suggesting that the inhibitory processes were not homogeneously distributed in the whole piriform cortex. Functional implications are discussed. PMID- 8713448 TI - Functional demarcation of a border between areas V6 and V6A in the superior parietal gyrus of the macaque monkey. AB - We have compared physiological data recorded from three alert macaque monkeys with separate observations of local connectivity, to locate and characterize the functional border between two related but distinct visual areas on the caudal face of the superior parietal gyrus. We refer to these areas as V6 and V6A. The occupy almost the entire extent of the anterior bank of the parieto-occipital sulcus, V6A being the more dorsal. These two areas are strongly interconnected. Anatomically, we have defined the border as the point at which labelled axon terminals first adopt a recognizably 'descending' pattern in their laminar characteristics, after injections of wheatgerm agglutinin-horseradish peroxidase into the dorsal half of the gyrus (in presumptive V6A). A similar principle was used to recognize the same border by the pattern of input from area V5, except that in this case the relevant transition in laminar characteristics is that between an 'intermediate' pattern (in V6) and an 'ascending' pattern (in V6A). V6A was found to be distinct from V6 in a number of its physiological properties. Unlike V6, it contains visually unresponsive cells as well as units with craniotopic receptive fields ('real-position' cells), units tuned to very slow stimulus speeds, units with complex visual selectivities and units with activity related to attention. V6A was also found to have a larger mean receptive field size and scatter than V6. By contrast, response properties related to the basic orientation and direction of moving bar stimuli were indistinguishable between V6 and V6A, as was the influence of gaze direction on cell activity in the two areas. Two-dimensional maps of the recording sites allowed reconstruction of the V6/V6A border. For comparison, the anatomical results were rendered on two dimensional maps of identical format to those used to summarize the physiological data. After normalizing for relative size, the physiological and connectional estimates of the border between V6 and V6A were found to coincide, at least within the range of individual variation between hemispheres. An architectonic map in the same format was also made from a hemisphere stained for myelin and Nissl substance. Area PO, defined by its general density of myelination was not distinct in this material, but several architectural features were traceable and one of these was also found to approximate the V6/V6A border. The particular criteria that distinguish V6 from V6A differ from a recent description of areas PO and POd in the Cebus monkey; we believe it most likely that PO and POd together may correspond to V6. PMID- 8713449 TI - Ontogenetic development of 5-HT1D receptors in human brain: an autoradiographic study. AB - The pattern of pre- and postnatal appearance of 5-HT1D receptors throughout the different areas of the human brain was studied by quantitative in vitro autoradiography, using [125I]GTI (serotonin O-carboxymethyl-glycyl [125I]tyrosinamide) as a ligand. The anatomical distribution of 5-HT1D receptors in neonatal, infant and children's brain was in good agreement with that observed in the adult, the basal ganglia and substantia nigra being the most intensely labelled areas. The development of these receptors throughout the human brain was mainly postnatal: low densities of [125I]GTI binding sites were observed at the fetal/neonatal stage in most regions analyzed, in contrast with the high levels of labelling found in infant and children's brains. Indeed, in a number of regions, including the globus pallidus, substantia nigra and visual cortex, a peak of overexpression of 5-HT1D receptors was observed in the first decade of life. Such overexpression could support a regulatory role for 5-HT1D receptors in advanced periods of the CNS developmental process. Our results also indicate that the administration of drugs acting on 5-HT1D receptors during the early postnatal period of life could result in modifications of their properties, as these receptors are already functional in this period. PMID- 8713450 TI - Differential distribution of MAP1a and aldolase c in adult mouse cerebellum. AB - MAP1a is a microtubule-associated protein with an apparent molecular weight of 360 kDa that is found in the axonal and dendritic processes of neurons. Two monoclonal anti-MAP1a antibodies anti-A and anti-BW6, revealed different epitope distributions in the adult mouse cerebellum. Anti-A stained Purkinje and granule cells uniformly throughout the cerebellum. In contrast, anti-BW6 selectively stained the dendriites of a subset of Purkinje cells, revealing parasagittal bands of immunoreactivity in the molecular layer. The compartmentation of the BW6 epitope was compared to the Purkine cells as revealed by immunostaining with anti zebrin II, a well known antigen expressed selectively by bands of Purkinje cells. The anti-BW6 staining pattern was complementary to the zebrin II bands, the zebrin II- Purkinjke cells having BW6+ dendrites. These results demonstrate that MAP1a is present in two forms in the mouse cerebellum, one of which is segregated into parasagittal bands. This may indicate a unique MAP1a isoform or may reflect differences in the metabolic states of Purkinje cell classes, and regional differences in their functions. PMID- 8713451 TI - Characterization of excitatory amino acid neurotoxicity in N-methyl-D-aspartate receptor-deficient mouse cortical neuronal cells. AB - Roles and mechanisms of N-methyl-D-aspartate (NMDA) receptors in glutamate neurotoxicity were investigated in cultures of NMDA receptor-deficient cortical neuronal cells. Mutant mice lacking a functional NMDA receptor were generated by gene targeting of the NR1 NMDA receptor subunit. Cortical neuronal cells prepared from wild-type NR1+/+, heterozygous NR1+/- and homozygous mutant NR1-/- mice at 15-17 days of gestation grew indistinguishably from each other. Brief exposures (5 min) of both NR1+/+ and NR1+/- neuronal cells to glutamate or NMDA, but not kainate or alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA), resulted in widespread neuronal degeneration by the following day. In contrast, neither glutamate nor NMDA treatment caused neuronal degeneration in NR1-/- cells, indicating that NMDA receptors are responsible for rapidly triggered glutamate neurotoxicity. The above four compounds were all effective in inducing the death of NR1+/+ and NR1+/- neuronal cells after prolonged exposure (20-24 h). However, NMDA had no neurotoxic effects on NR1-/- cells, although the other three compounds wer neurotoxic with potencies comparable to those for NR1+/+ and NR1+/- cells. The AMPA and kainate receptors are thus sufficient for inducing slowly triggered glutamate neurotoxicity. Brief exposure of a mixed population of NR1+/+ and NR1-/- neuronal cells to NMDA selectively killed the NMDA receptor-expressing cells without any appreciable effects on neighbouring NMDA receptor-deficient cells. This finding further supports a direct and indispensable role for NMDA receptors in NMDA-evoked neuronal cell death. PMID- 8713452 TI - Immunocytochemical evidence that glutamate is a neurotransmitter in the cochlear nerve: a quantitative study in the guinea-pig anteroventral cochlear nucleus. AB - The large so-called type I afferents of the cochlear nerve carry the majority of the auditory input from the cochlea to the cochlear nuclei in the brainstem. These fibres are excitatory and previous studies have suggested they may use glutamate as their neurotransmitter. In the present investigation therefore, antibodies to glutamate and to the glutamate precursor, glutamine, were applied to resin sections of perfusion-fixed brains and of in vitro brain slices subjected to depolarizing levels of potassium before fixation to study glutamate handling and synaptic release. Ultrathin sections were labelled by the immunogold technique, and the immunoreactivity was quantified by recording the density of gold particles over the various tissue profiles. Non-primary, presumably inhibitory, terminals and glial processes were used as reference structures. The cochlear primary terminals proved to be strongly immunoreactive for glutamate. The density of glutamate labelling was higher in primary terminals than in non primary ones, and lowest in glial processes. The ratio between the mean glutamate and glutamine labelling densities was also higher in primary terminals than in non-primary ones, and lowest in glial processes in each case. In the primary terminals, the glutamate immunoreactivity was higher over vesicle-containing regions than over vesicle-free regions, whilst glutamine was evenly distributed throughout. The in vitro brain slices showed a potassium-induced, partly calcium dependent depletion of glutamate from the primary terminals but not from the non primary ones. These observations strongly support the conclusion that glutamate is a neurotransmitter of type I cochlear afferents. PMID- 8713453 TI - Responses of neurons of the nucleus of the optic tract and the dorsal terminal nucleus of the accessory optic tract in the awake monkey. AB - The nucleus of the optic tract (NOT) and the dorsal terminal nucleus of the accessory optic tract (DTN) are essential nuclei for the generation of slow-phase eye movements during horizontal optokinetic nystagmus. We recorded from 101 neurons (all directionally selective) in four NOT/DTN of three trained and behaving rhesus monkeys. Neuronal activity increased when stimuli moved ipsiversively with respect to the recording site and decreased below spontaneous activity when stimuli moved contraversively. While the monkey fixated a small spot, some NOT/DTN neurons did not respond at all to the retinal image slip of a whole-field random dot pattern; others showed a monotonic increase of activity to increasing velocities of that stimulus. The velocity range tested was up to 100 degrees/s. During the execution of optokinetic nystagmus, 39 of 73 cells tested showed a velocity-tuned response with an average optimum at 21 degrees/s retinal image slip. Following saccades during optokinetic nystagmus (quick phases), the NOT/DTN neuronal activity briefly attained the level of spontaneous activity, as predicted from the velocity selectivity during optokinetic nystagmus. Immediately upon cessation of optokinetic stimulation in the preferred direction, NOT/DTN activity returned to the spontaneous level and did not reflect the ongoing optokinetic afternystagmus in darkness. Most NOT/DTN neurons displayed direction selectivity also during smooth pursuit. Twenty-one of 50 cells tested (42%) always responded to the retinal slip of the target (target velocity cells), 16 cells (32%) responded to the retinal slip of the background (background velocity cells), and 13 cells (26%) did not respond at all during smooth pursuit. We conclude from our results that the NOT/DTN is an essential structure for the processing of the direction and speed of retinal image slip. This information is then used for the generation and maintenance of slow eye movements, preferentially during horizontal optokinetic nystagmus but also during pursuit eye movements. PMID- 8713454 TI - The effects of lesions to the mammillary region and the hippocampus on conditional associative learning by rats. AB - Rats with extensive lesions to the mammillary body region, the hippocampus, or rats which had received a control operation were trained postoperatively on two visuo-spatial conditional associative learning tasks in which they had to learn to associate spatial cues with particular visual/auditory stimuli. The animals were subsequently trained on a spatial working memory task, the eight-arm radial maze. Rats with lesions to the mammillary body region were able to acquire the conditional associative learning tasks at a rate comparable to that of operated control animals, whereas those with hippocampal lesions were not. By contrast, rats with a lesion of the mammillary body region or the hippocampus were significantly impaired in comparison with the operated control animals in the radial maze. The findings suggest that lesions to the mammillary body region impair spatial working memory without affecting the capacity to associate particular exteroceptive cues with spatial locations. PMID- 8713455 TI - Glutamate depresses release by activating non-conventional glutamate receptors at crayfish nerve terminals. AB - The present study shows that release of glutamate from crayfish nerve terminals is inhibited at low depolarizing current pulses by glutamate, N-methyl-D aspartate (NMDA) and quisqualate. These agonists elicit inhibitory effects at concentrations as low as 10(-8) M (quisqualate) and 10(-7) M (glutamate and NMDA). The NMDA-mediated inhibition is blocked by (+/-)-2-amino-5 phosphonovaleric acid (APV). The quisqualate-mediated inhibition is blocked by 6 cyano-7-nitroquinoxaline-2,3-dione (CNQX). Both CNQX and APV are needed to block glutamate-mediated inhibition. The inhibition of release is not accompanied by a detectable change in presynaptic membrane conductance at the secondary branch. Using fura-2, Ca2+ accumulation during repetitive stimulation (100 Hz) was monitored in single release boutons. Inhibition of release, elicited by 10(-4) M glutamate, was not associated with a reduction in the accumulation of Ca2+. We show that the glutamate released from a single or a few release boutons during normal activity acts similarly to glutamate added externally, i.e. it inhibits its own release. PMID- 8713456 TI - Distribution and sites of synthesis of NTT4, an orphan member of the Na+/Cl(-) dependent neurotransmitter transporter family, in the rat CNS. AB - The distribution and sites of synthesis in rat CNS of NTT4, a novel orphan member of the Na+/Cl(-)-dependent neurotransmitter transporter family, were determined by immunohistochemistry and hybridization histochemistry. Antibodies raised against recombinant fusion proteins, corresponding to residues of NTT4, and 35S labelled oligodeoxyribonucleotide probes, were used to delineate the cellular distribution of the transporter at the protein and mRNA levels. High levels of immunoreactivity (mainly in the neuropil) were found in the olfactory bulb, cerebral cortex, striatum, hippocampus, thalamus, substantia nigra, pontine nuclei, cerebellum and spinal cord. The lowest levels were associated with the lateral hypothalamic area and deep mesencephalic nuclei. In situ hybridization signals correlated well with the immunoreactivity, and demonstrated a widespread distribution of NTT4 transcripts exclusively in neurons. NTT4 transcripts appeared widely codistributed with the N-methyl-D-aspartate receptor subunit 1 (1 4b), i.e. spliced variants characterized by a common 5' 63 bp insertion. These results indicate that the transporter was associated with neuronal processes in specific glutamate innervated CNS regions. Although the substrate transported by NTT4 remains unknown, our findings suggest a possible role for this carrier protein in glutamate/glycine neurotransmission. PMID- 8713457 TI - DCG-IV selectively attenuates rapidly triggered NMDA-induced neurotoxicity in cortical neurons. AB - Molecular cloning has revealed the existence of at least eight subtypes of metabotropic glutamate receptors (mGluRs). We examined the effect of (2S,1'R,2'R,3'R)-2-(2,3-dicarboxycyclopropyl)glycine (DCG-IV), a selective agonist of the mGluR 2/3 subtype, on excitotoxicity in mouse cortical cell cultures. Addition of DCG-IV to the exposure medium partially attenuated the rapidly triggered excitotoxic death induced by a 5 min exposure to 200 microM NMDA. This neuroprotective effect was reversed by coapplication of alpha-methyl-4 carboxyphenylglycine (MCPG), an antagonist of mGluRs, by pertussis toxin pretreatment and also by preincubation with dibutyryl cAMP, a stable analogue of cAMP. These results suggest that the activation of mGluR 2/3 is neuroprotective in our system. However, DCG-IV did not attenuate the slowly triggered neuronal death induced by 24 h exposure to low concentrations of NMDA, alpha-amino-1,3 cyclopentanedicarboxylic acid (AMPA) or kainate. The failure of DCG-IV to block slowly triggered NMDA neurotoxicity is likely due to weak NMDA agonist activity, as demonstrated in whole-cell recording. PMID- 8713458 TI - Further observations on the role of nitric oxide in the feline lateral geniculate nucleus. AB - We have examined the responses of a population of 77 cells in the dorsal lateral geniculate nucleus (dLGN) of the anaesthetized, paralysed cat. Here the synthetic enzyme for the production of nitric oxide, nitric oxide synthase, is found only in the presynaptic terminals of the cholinergic input from the brainstem. In our hands, iontophoretic application of inhibitors of this enzyme resulted both in significant decreases in visual responses and decreased responses to exogenous application of NMDA, effects which were reversed by coapplication of the natural substrate for nitric oxide synthase, L-arginine, but not the biologically inactive isomer, D-arginine. Nitroprusside and S-nitroso-N-acetylpenicillamine (SNAP), nitric oxide donors, but not L-arginine, were able to increase markedly both spontaneous activity and the responsiveness to NMDA application. Furthermore, SNAP application facilitated visual responses. Responses of cells in animals without retinal, cortical and parabrachial input to the LGN suggest a postsynaptic site of action of nitric oxide. This modulation of the gain of visual signals transmitted to the cortex suggests a completely novel pathway for nitric oxide regulation of function, as yet described only in primary sensory thalamus of the mammalian central nervous system. PMID- 8713459 TI - Neurotensin excitation of serotonergic neurons in the dorsal raphe nucleus of the rat in vitro. AB - Neurotensin-containing terminals and radioligand binding sites are present in the dorsal raphe nucleus. The purpose of this study was to test, in brain slices containing this nucleus, the effect of neurotensin on the electrical activity of serotonergic neurons. In extracellular recordings, the cells were identified by the ability of the alpha 1-adrenoceptor agonist phenylephrine to induce firing, and serotonin to reduce this effect. After washout of phenylephrine, neurotensin (10 nM to 10 microM) induced a concentration-dependent increase in the firing rate of serotonergic neurons (EC50 = 142 nM; maximum effect approximately 1 microM). The neurotensin excitation, which was mimicked by neurotensin fragments 8-13 but not neurotensin peptide fragment 1-8 and selectively blocked by SR 48692 (100 nM), was observed mainly in the ventral part of the nucleus. Most serotonergic neurons showed marked desensitization to neurotensin, even at low concentrations. The neurotensin response was occluded by supramaximal concentrations of phenylephrine. In intracellular recordings using KCl-containing electrodes, neurotensin induced an inward current associated in some cases with a decrease in apparent input conductance. In conclusion, neurotensin was found to have an excitatory action on serotonergic neurons in the ventral part of the dorsal raphe nucleus, an effect which could be subject to desensitization and was occluded by phenylephrine. This occlusion phenomenon may be important for the physiological role of neurotensin in the dorsal raphe nucleus. PMID- 8713460 TI - Rab3 proteins and SNAP-25, essential components of the exocytosis machinery in conventional synapses, are absent from ribbon synapses of the mouse retina. AB - GTP-binding rab proteins, present in synaptic vesicles and endocrine secretory granules, have been shown to be involved in the control of regulated exocytosis. We found rab3 proteins in immunoblots of diverse areas of the mouse central nervous system (spinal cord, olfactory bulb, hippocampus, cerebellum and neocortex). Immunohistochemical observations at light- and electron-microscopical levels in the hippocampus and other areas revealed rab3 proteins in virtually all synaptic fields and terminals of the areas investigated. In the retina, rab3A immunoreactivity was confined to the inner and outer plexiform layers. Ultrastructural examination revealed that rab3A was present in conventional terminals in the inner plexiform layer and in horizontal cell processes of the outer plexiform layer. In contrast ribbon synapses, which play a key role in transferring information from the photoreceptor cells to the central nervous system, were immunonegative. We also tested whether other proteins of the rab3 family are present in ribbon synapses. However, using an antibody recognizing rab3B and rab3C in addition to rab3A, we found no immunoreactivity in these synapses. Interestingly, we observed also no immunoreactivity for synaptosomal associated protein 25 (SNAP-25) in ribbon synapses, but conventional synapses and horizontal cell processes were heavily stained. Our data show that the known rab3 and SNAP-25 isoforms, which are components of the secretory apparatus of conventional synapses, are absent from ribbon synapses of the retina. Our observations suggest different mechanisms of transmitter exocytosis in conventional and ribbon terminals. PMID- 8713461 TI - Differential oscillatory properties of cholinergic and noncholinergic nucleus basalis neurons in guinea pig brain slice. AB - Evidence has suggested that the nucleus basalis magnocellularis has the potential to influence the functional state of the cerebral cortex through topographically organized, widespread projections of the cholinergic cells in that nucleus. It has also been shown that, in addition to the cholinergic neurons, other non cholinergic magnocellular basal forebrain neurons, some of which have been identified as gamma-aminobutyric acid-ergic, project into the cerebral cortex and thus may also participate in the modulation of its activity. We have performed a comparative study of the intrinsic rhythmic properties of immunohistochemically and morphologically characterized choline acetyltransferase (ChAT)-positive and ChAT-negative cells of the nucleus basalis by means of intracellular recordings in guinea pig brain slices. Our results demonstrate that relatively large, multipolar cholinergic and non-cholinergic neurons each display differential voltage-dependent properties that allow them to discharge rhythmically in spike bursts and spike clusters, respectively, at low frequencies (< 10 Hz). Cholinergic cells display bursts of 2-4 action potentials (at approximately 200 Hz) riding on low-threshold spikes recurring at a low frequency (< 5 Hz) when depolarized from a membrane potential more negative than -55 mV and display low frequency (< 10-15 Hz) tonic firing when depolarized from a more positive level. In contrast, non-cholinergic cells fire in a unique mode, displaying non-adapting clusters of spikes interspersed with rhythmic subthreshold membrane-potential oscillations when depolarized from levels less negative than -55 mV. The spike clusters repeat rhythmically at relatively low frequencies (2-10 Hz). The intracluster spiking frequency is relatively high and coincides approximately with that of the intervening membrane-potential oscillations (approximately 20-70 Hz). The cluster frequency of the non-cholinergic cells corresponds, in the same manner as the burst frequency of the cholinergic cells, to a delta (1-4 Hz) or theta (4-10 Hz) range of activity, whereas the intra-cluster and tonic spike frequencies of the non-cholinergic cells correspond to high beta to gamma ranges of electroencephalographic activity (19-30 Hz and 30-60 Hz, respectively). We propose that the different modes of oscillatory firing by the cholinergic and non cholinergic basal forebrain cell populations could collectively contribute to the rhythmic modulation of slow and fast rhythms within the cerebral cortex. PMID- 8713462 TI - Potassium channel mRNA expression in prevertebral and paravertebral sympathetic neurons. AB - The expression of eighteen different voltage-activated potassium channel genes in rat sympathetic ganglia was quantitatively analysed using an RNase protection assay. Eleven alpha-subunit genes and two beta-subunit genes were expressed in sympathetic ganglia. The relative level of potassium channel mRNA expression was compared between the superior cervical ganglion (SCG) and two preverteabral sympathetic ganglia, the coeliac ganglion (CG) and the superior mesenteric ganglion (SMG). Four mRNAs were differentially expressed: Kv1.2, Kv1.4, Kv2.2 and Kv beta 1. Transcripts from all four genes were more abundant in the prevertebral ganglia. From comparisons with previous electrophysiological studies it was concluded that genes encoding the channels underlying the M-current and D2 current, which are both prominent in sympathetic neurons, have yet to be identified. It was also concluded that members of the Kv4 family are likely to underlie the low-threshold A-current in sympathetic neurons. PMID- 8713463 TI - Mechanisms for synchronous calcium oscillations in cultured rat cerebellar neurons. AB - Removal of Mg2+ caused oscillations of the cytosolic Ca2+ concentration ([Ca2+]i) and the membrane potential in cultured cerebellar granule neurons. Oscillations of [Ca2+]i were synchronous in all the cells, and were restricted to the neurons (immunocytochemically identified) that responded to exogenous N-methyl-D aspartate (NMDA). Oscillations were blocked by Ca2+ removal, nickel, NMDA receptor antagonists, omega-agatoxin IVA, tetrodotoxin, sodium removal and gamma aminobutyric acid, but not by dihydropyridines, omega-conotoxin M VIIA or by emptying the intracellular Ca2+ stores with thapsigargin or ionomycin. The upstroke of the [Ca2+]i oscillations coincided in time with an increase in manganese permeability of the plasma membrane. Propagation of the [Ca2+]i wave followed more than one pathway and the spatiotemporal pattern changed with time. Membrane potential oscillations consisted of transient slow depolarizations of approximately 20 mV with faster phasic activity superimposed. We propose that the synchronous [Ca2+]i oscillations are the expression of irradiation of random excitation through a neuronal network requiring generation of action potentials and functional glutamatergic synapses. Oscillations of -Ca2+-i are due to cyclic Ca2+ entry through NMDA receptor channels activated by synaptic release of glutamate, which requires Ca2+ entry through P-type Ca2+ channels activated by action potentials at the presynaptic terminal. PMID- 8713464 TI - TGF-beta rescues target-deprived preganglionic sympathetic neurons in the spinal cord. AB - Transforming growth factors beta (TGF-beta), a family of pleiotropic cytokines, are widely distributed in the developing and adult nervous system. In order to further determine the neural functions of TGF-beta, we have localized the TGF beta isoforms 1, 2 and 3 in the adult rat adrenal medulla and studied the neuroprotective capacity of one representative family member, TGF-beta 2, for those spinal cord neurons which innervate adrenal chromaffin cells and which die after destruction of the adrenal medulla. Unilateral electrothermal destruction of the adrenal medulla led to the disappearance of 25% of sympathetic preganglionic neurons, which are located in the intermediolateral (IML) column of thoracic spinal cord segments 7-10 and can be selectively marked by NADPH diaphorase. The neurons which disappeared following adrenomedullectomy constitute the full set of neurons that innervate the adrenal medulla. Implantation of gelfoam soaked with 0.5 micrograms TGF-beta 2 into the adrenal wound cavity rescued all spinal cord neurons in the IML ipsilaterally to the lesioned side. Cytochrome c was not effective. Injections of [125I]TGF-beta 2 into the adrenal medulla did not result in retrograde transport and subsequent labelling of spinal cord neurons, suggesting that TGF-beta may exert its neuroprotective actions by indirect mechanisms. TGF-beta applied to cultured adrenocortical cells did not overtly increase the amount of mRNA for fibroblast growth factor-2, an established trophic molecule for sympathetic preganglionic spinal cord neurons. The mechanisms by which TGF-beta exerts its neurotrophic effect are therefore unclear. Even so, our data provide the first evidence that TGF-beta may play an important role in vivo in the control of maintenance of a population of spinal cord neurons. PMID- 8713465 TI - Cholinergic modulation of the action potential in rat hippocampal neurons. AB - The cholinergic input to the hippocampus from the medial septum is important for modulating hippocampal activity and functions, including theta rhythm and spatial learning. Neuromodulation by transmitters in central nervous system neurons usually affects cell excitability by modifying the membrane potential, discharge pattern and spike frequency. Here we describe another type of neuromodulation: changing the action potential waveform. During intracellular recordings from CA1 pyramidal cells in hippocampal slices from rats, the cholinergic agonist carbachol caused several reversible changes in the action potential: low doses (2 microM) caused an increase in spike duration; high doses (10-40 microM) or long lasting applications also reduced the spike amplitude and rate of rise, and raised the spike threshold. These effects are similar to those of metabotropic glutamate receptor agonists or phorbol esters, both of which activate protein kinase C. The effects were blocked by the muscarinic antagonist atropine, and were prevented by Ca(2+)-free medium and by Ca(2+)-channel blockers. However, the cholinergic spike modulation was not occluded or mimicked by blocking the Ca(2+) dependent K+ currents IC or IAHP, suggesting that these K+ currents are not involved in the modulation. We conclude that muscarinic receptor activation modulates the action potential in CA1 pyramidal cells via a Ca(2+)-dependent mechanism, possibly involving protein kinase C. This modulation and the similar effects mediated by metabotropic glutamate receptors to our knowledge provide the only examples of neuromodulation of the action potential in the vertebrate central nervous system-a form of modulation known to regulate Ca2+ influx and transmitter release, and to mediate synaptic plasticity and learning in invertebrates. PMID- 8713466 TI - Unique distribution of somatostatin-immunoreactive cells in the retina of the tree shrew (Tupaia belangeri). AB - Somatostatin-like immunoreactive cells in the tree shrew retina were studied with the monoclonal antibody S8 against the neuropeptide somatostatin 14. As in some other mammals, immunoreactive somata are exclusively found in the ganglion cell layer. Immunoreactive processes form a sparse main plexus in the inner plexiform layer near the border of the inner nuclear layer; fewer additional processes are found closer to the ganglion cell layer. With retrograde labelling of retinal ganglion cells by injections of the tracer Fast Blue into the superior colliculus and lateral geniculate body and counterstaining of the retinae with S8, approximately 5% of the immunoreactive somata were double-labelled at any retinal location. The vast majority of somatostatin-like immunoreactive cells are thus displaced amacrine cells. Their somata are distributed over the entire retina. Their population density is highest in the temporal retina, with peak densities of approximately 5000 cells/mm2 near the central area and a steep density gradient. In the remaining retina densities are 200-400 cells/mm2, falling to approximately 100 cells/mm2 at the retinal margins. This is in stark contrast to the somatostatin-like immunoreactive cells in other mammalian retinae which have densities of 10-40 cells/mm2 and are confined to restricted retinal regions (inferior retina and/or retinal margin). PMID- 8713467 TI - Differential localization of AMPA glutamate receptor subunits in the two segments of the globus pallidus and the substantia nigra pars reticulata in the squirrel monkey. AB - The subthalamic nucleus has long been known as the main source of glutamatergic afferents to the pallidum and the substantia nigra in primates. Recent findings showed that the excitatory effects induced by the subthalamic nucleus in pallidal cells are mediated through the activation of non-NMDA receptors in the rat. The objective of the present study was to analyse the distribution of alpha-amino-3 hydroxy-5-methyl-4-isoxazole-propionate (AMPA) glutamate receptor subunits in the external pallidum (GPe), the internal pallidum (GPi) and the substantia nigra pars reticulata (SNr) in squirrel monkeys (Saimiri sciureus). This was achieved by means of immunohistochemistry using antibodies raised against the GluR1 and the GluR2/3 subunits of the AMPA receptor. Our results show that all neuronal perikarya in GPe and GPi display immunoreactivity for GluR2/3 subunits whereas GluR1 is confined exclusively to cells in the GPe. The proportion of GluR1 immunoreactive neurons is not uniform throughout the rostrocaudal extent of GPe; in the rostral third all GPe cells display GluR1 immunoreactivity, whereas in the caudal third the proportion of GluR1-positive cells decreases to 50%. The intensity of GluR1 immunostaining associated with GPe cells is lower than that associated with neighbouring large-sized neurons in the nucleus basalis of Meynert. In contrast to GPi cells, the neurons in the SNr display immunoreactivity for both GluR1 and GluR2/3 subunits. In conclusion, our results provide the first evidence for a different distribution of the GluR1 subunit of the AMPA receptors in the two segments of the globus pallidus in monkeys. These findings imply that the control of the basal activity of GPe and GPi cells by the subthalamic nucleus is exerted via the activation of AMPA receptors composed of different subunits. These data reinforce the view that the two segments of the globus pallidus are different entities that possess their own functional characteristics in primates. PMID- 8713468 TI - Prolactin as an immunoreactive agent. AB - Prolactin (PRL), an endocrine hormone from the anterior pituitary, is also synthesized and secreted by activated immunocytes, mostly T cells and thymocytes. In vitro PRL acts as an autocrine or paracrine growth factor which regulates the proliferation of previously stimulated immunocompetent cells. PRL acts through its specific cell surface receptors expressed on different immunocytes including T cells, B cells, monocytes, NK cells and neutrophils. In vivo the immunoregulatory role of PRL is still the subject of intense investigation, especially in the conditions of stress and reproduction. The conflicting results suggest that PRL concentration, sex hormones and some other factors influence the immunomodulatory role of PRL. PMID- 8713469 TI - Genetic approaches to the study of cytokine regulation of mucosal immunity. AB - Efforts to design effective mucosal vaccines have been hampered by an incomplete understanding of factors controlling the development of mucosal immunity. It is now clear, however, that T cell-derived cytokines play a major role. Recent developments in 'gene knockout' technology have allowed the generation of strains of mice in which particular genes have been inactivated. The availability of mice rendered deficient for production of Th2 cytokines has facilitated studies of the induction and development of mucosal immune responses in the absence of these factors. We have used several genetic approaches, including cytokine-deficient mice and recombinant vectors constructed to express genes for a range of different cytokines, to demonstrate the importance of these factors in the mucosa. Such genetic approaches appear to represent powerful tools for in vivo studies of the influence of cytokines in mucosal immunoregulation. PMID- 8713470 TI - The ecology and pathology of Epstein-Barr virus. AB - Epstein-Barr virus achieves its ubiquitous and uniform epidemiological distribution by a dual strategy of latency to guarantee lifelong persistence and intermittent replication to guarantee transmission. These two functions appear to dictate residence in different cell types: latency in B lymphocytes and replication in epithelial cells. Both of these cell compartments are potential sites for EBV-associated malignancies. PMID- 8713471 TI - Characterization of immunorelated peptides to porcidin P1. AB - Porcidin P1, an antimicrobial peptide purified from the granules of porcine polymorphonuclear neutrophils (PMN) using ultrafiltration and reverse phase high performance liquid chromatography (RP-HPLC), was covalently conjugated to BSA and used to generate monospecific polyclonal ascites. Antibodies raised against porcidin P1 were covalently coupled to an Affi-gel Hz affinity column and used for immunoaffinity chromatography of peptides from porcine PMN cell extract. Eleven immunorelated peptides were eluted from the column from neutrophil cell extracts and purified to homogeneity by HPLC. The molecular weights of the immunorelated peptides were determined by mass spectral analysis and ranged in size from 1.91 to 10.65 kDa. Of the 11 immunorelated peptides which were bound to the affinity column, only six peptides were recognized by the anti-porcidin antibodies after HPLC purification. Three immunoreactive peptides displayed potent antibacterial activity towards Staphylococcus aureus and Escherichia coli, reducing viability by as much as 99.9% (> 3 log reduction in CFU) when 5 mu g/mL of each purified peptide was used. The polyclonal monospecific antibodies also reacted with proteins from ovine and human PMN, illustrating possible structural relationships between small antibacterial peptides from the different species. PMID- 8713472 TI - Altered calcium signal transduction in B-1 malignant cells. AB - Lymphocyte proliferation is guided by receptor-mediated signal transduction pathways that dictate the immunological response/clonality of that cell. We have previously reported that NZB-derived malignant B-1 cells, which serve as a murine model for human chronic lymphocytic leukaemia, demonstrate altered expression of surface IgM and CD45 signalling molecules, and a failure to proliferate following membrane IgM stimulation. To examine receptor-mediated cytosolic calcium (Cai) signalling in B cell leukaemia, we studied IgM-induced Cai responses in malignant B-1 cells and B cells from non-leukaemic mice. Basal Cai was slightly lower in malignant B-1 cells than in non-leukaemic cells. Anti-IgM stimulation induced a sustained increase in Cai to levels 1.3-fold greater than basal Cai in conventional B cells. In contrast, leukaemic B-1 cells demonstrated a sharp but transient rise in Cai followed by a gradual increase to levels 2.3-fold greater than basal [Ca]i Ca influx from extracellular sources contributed to the early and late Cai signal in both sets of cells. Pre-incubation (2-30 min) with anti CD45 had no effect on basal Cai or the anti-IgM Cai signal in B cells, but reduced the Cai transient in malignant B-1 cells. Additional experiments characterized the effects of phosphorylation/dephosphorylation events on the Cai profile following anti-IgM stimulation. Protein tyrosine kinase inhibitors decreased the anti-IgM-induced Cai transient in malignant B-1 cells by 80%, but only moderately affected (40%) of the Cai response in non-leukaemic B cells. Protein tyrosine phosphatase inhibitors and protein kinase C (PKC) activators attenuated the Cai response to the same degree in normal and leukaemic B cells. These results show that Cai signalling differs widely between non-malignant B cells and malignant B-1 cells, and that tyrosine phosphorylation and CD45 modulation of IgM signalling are involved in the altered Cai responses in malignant B-1 cells. PMID- 8713473 TI - Expression of rabbit C-reactive protein in transgenic mice. AB - C-reactive protein (CRP) is a prototypic acute phase reactant in humans and rabbits whose serum concentration can increase up to 1000-fold following an acute inflammatory stimulus. CRP binds to many phosphate ester-containing compounds including phosphorylcholine, nucleotides, chromatin and snRNP. To examine the in vivo function of this protein, we produced transgenic mice capable of significant CRP synthesis. In contrast to most other vertebrates, mice synthesize CRP in only trace amounts. The transgenic animals express rabbit CRP from either the phosphoenolpyruvate carboxykinase promoter (PEPCK-CRP) or the mouse metallothionein I promoter (MT-CRP). Manipulating the diet in one of the PEPCK CRP lines led to a rise in serum CRP levels from < 5 mu g/mL to 100-200 mu g/mL over a period of 2 days. The two MT-CRP lines examined expressed CRP constitutively which could be further elevated 2-4-fold following an inflammatory stimulus. Transgenic CRP bound phosphorylcholine was pentameric, had a circulating half-life of 30-60 min and was capable of activating mouse complement when bound to a ligand. We conclude that these transgenic lines express CRP with many of the properties of authentic rabbit CRP, and that the expression of CRP can be controlled to be dependent or independent of the acute phase response. PMID- 8713474 TI - The T cell antigen receptor beta chain interacts with the extracellular domain of CD3-gamma. AB - Selective pairwise interactions between a number of CD3 chains and the clonotypic T cell antigen receptor (TCR-alpha, -beta) chains have recently been reported. What still remains unanswered is the site of interaction between TCR-beta and CD3 gamma chains. To examine the region of interaction between TCR-beta and CD3-gamma chains, a variety of genetically altered TCR-beta and CD3-gamma chains were constructed using recombinant cDNA techniques. Non-T cells (COS-7) were transfected with cDNA constructs, metabolically labelled, and immunoprecipitates were analysed for assembly using non-equilibrium pH gel electrophoresis (NEPHGE)/SDS-PAGE. The results demonstrated that assembly between TCR-beta and CD3-gamma chains was localized to their extracellular domain. These findings, when coupled with the information on pairwise interactions and formation of higher order subcomplexes, extend our model of the structure of the TCR complex. PMID- 8713475 TI - Dendritic cell presentation of PPD and 19 kDa protein of Mycobacterium tuberculosis and emergent T helper cell phenotype. AB - Protection against infection with Mycobacterium tuberculosis is preferentially associated with the development of the T helper 1 subset, IFN-gamma production and a cell-mediated response, rather than with T helper 2 cells, 4 (IL-4) and antibody production. The type of APC interacting with T cells responsive to mycobacterial peptides may influence which of these responses predominates. This investigation focuses on the role of dendritic cells (DC) because they are the most potent APC in both primary and recall immune responses. Our results show that splenic DC-enriched suspensions prepared from C57BL/6 mice and pulsed with either purified protein derivative (PPD) or the immunodominant 19 kDa protein from M. tuberculosis, can activate antigen-primed T cells in vitro, whereas spleen cell suspensions depleted of DC cannot. DC pulsed with PPD or 19 kDa antigen are able to prime naive T cells in vivo. Supernatants collected from cultures containing T cells from mice injected with PPD-pulsed DC and then challenged in vitro with PPD-pulsed DC were found to contain more IL-2 and IFN gamma than those from control mice which received either DC or PPD alone. No such antigen-specific IFN-gamma response occurred if DC pulsed with 19 kDa were used in place of PPD-pulsed DC. IL-4 was not detected in any of the culture supernatants. We conclude that DC can induce production of cytokines associated with a protective immune response when presenting peptides derived from heterogeneous mycobacterial antigens but not when exposed to the single 19 kDa immunodominant protein. PMID- 8713476 TI - Hierarchy of T cell antigen receptor assembly. AB - The formation of partial complexes in an African green monkey kidney cell line (COS cells) has been used to examine assembly of newly synthesized T cell antigen receptor (TCR) chains. The identification of assembly interactions between multiple subunits and the formation of higher order subcomplexes has led to the development of a model for the hierarchy of subunit assembly leading to a complete TCR. These assembly interactions suggest likely nearest neighbour relationships in the assembling structure and probably reflect the quarternary structure of the TCR complex. This may be important when examining the association of other molecules with TCR proteins or when trying to discern structural components involved with signal transduction. PMID- 8713477 TI - Autoimmune diabetes: caught in the causality trap? AB - The lack of concordance between genotype and clinical diabetes has prompted a search for the infectious agent that precipitates this autoimmune disease. However, this approach may be misleading. It assumes that the disease-prone individuals that do not develop diabetes do not have autoimmunity. In the non obese diabetic (NOD) mouse, the genotype is a primary determinant of autoimmunity. Not all animals of the disease-prone genotype develop clinical disease; however, all have autoimmunity. This is expressed as a destructive or non-destructive process. Multiple pathways are open to the immune system and whether or not the immune response is destructive and leads to the development of clinical disease, appears to be a random process. If this is the case, the most important questions relating to autoimmune disease are not those concerning the 'causative' agents. Instead we should be asking what are the differences between pathways open to the immune system and what factors affect the probability that one or another pathway is finally selected? PMID- 8713478 TI - Depletion of IFN-gamma, CD8+ or Tcr gamma delta+ cells in vivo during primary infection with an enteric parasite (Trichostrongylus colubriformis) enhances protective immunity. AB - In order to examine the role of CD8+ and WCI+ T cells and of IFN-gamma in the development of protective immunity against infection with the enteric nematode parasite Trichostrongylus colubriformis in sheep, mAb were administered during induction of the immune response to deplete or neutralize these components. Protection against the primary and challenge infections were assessed by faecal egg count and total worm count. Prolonged administration of mAb recognizing IFN gamma and CD8+ resulted in significantly increased protection during the 6-week primary infection. CD8+ cells were depleted from blood but not intestinal mucosa. After injection of mAb (CC15) recognizing the surface antigen WCI, WCI+ and T cell receptor (Tcr) gamma delta+ cells were depleted from blood but not from enteric mucosa, and protection against challenge, although variable, was increased by up to 88%. It appears that CD8+ and WCI+/gamma delta+ cells and IFN gamma all retard the potential development of naturally-acquired immunity against the parasite. PMID- 8713479 TI - Role of the PS integrins in Drosophila development. AB - The PS1 and PS2 integrins of Drosophila are heterodimers of alphaPS1betaPS and alphaPS2betaPS subunits, respectively, with very strong structural similarity to vertebrate integrins. Cell transfection experiments indicate that the PS integrins are receptors for extracellular matrix components and are functionally analogous to vertebrate integrins. Matrix ligands from Drosophila tissues have been identified for PS1 and PS2 integrins, using transformed cells and a cell spreading assay. Mutations in all three subunit genes have been identified, and the phenotypes of mutants indicate that PS integrins are required for the proper morphogenesis of a number of embryonic tissues. Using methods to produce genetic mosaics and transformation of integrin transgenes into whole animals, integrin functions in adult morphogenesis also have been examined. In the pupal wing, integrins are critically required to maintain the connection between dorsal and ventral epithelia, and recent results suggest that in early pupal development, the integrins are acting as specific receptors, as opposed to a non-specific cell matrix glue. PMID- 8713480 TI - Contributions of an orthopteran to the understanding of neuronal pathfinding. AB - During the development of the nervous system neurons extend axons through a complex embryonic environment. To find a correct target, often located at a long distance, the neuronal growth cones travel along highly specific and stereotyped pathways. Proper neuronal pathfinding is thought to be accomplished by the specific interaction of receptors on the neuronal surface with molecular cues in the environment. We review the information obtained in an invertebrate model system, the grasshopper embryo, about the specific role of the cell surface in wiring the nervous system. PMID- 8713481 TI - Human melanoma integrins contribute to arrest and stabilization potential while flowing over extracellular matrix. AB - To form distant metastases, tumour cells must stabilize adhesive interactions that prevent detachment at secondary sites. Primary receptor-ligand interactions alone may not maintain prolonged adhesive contacts without secondary events that lead to adhesion stabilization. Computerized imaging methods enable us to examine various substrates for: (i) the wall shear adhesion threshold (WSAT), a measure of the dynamic adhesive potential of tumour cells; (ii) the number of tumour cells that adhered; and (iii) the adhesion stabilization lag time (ASLT) or length of time required for tumour cells to stabilize adhesive contacts capable of withstanding high wall shear force (up to 100 dynes/cm2). The relative WSAT ratios found were: wheat germ agglutinin (WGA) > laminin > fibronectin > vitronectin > collagen I > collagen IV > von Willebrand factor (vWF) (the greater the shear rate the higher the adhesive potential). The relative stabilization ratios found were as follows: laminin < fibronectin < vitronectin < collagen IV < collagen I < vWF < WGA (shorter times correlate with greater stabilization potential). Stabilization data using fibronectin as a substrate correlated the best with metastatic potential. Using three melanoma lines of different metastatic potential semiquantitative reverse transcriptase-polymerase chain reaction (PCR) showed a two- to four-fold increase in alpha1, alpha3, alpha4, alpha5, alpha6, and ICAM-1 in the highly metastatic 70W cells compared to the MeWo and non-metastatic 3S5 melanoma cells. There were no differences in alphav, beta1 and beta3 levels among the three melanoma lines, and PCR products for alphaIIb, alpha2, CD36, or ICAM-2 were not detected. The 70W cells also had higher levels of alphax and beta2 (CD11/CD18 and p150 leukocyte antigen) than either the MeWo or 3S5 cells. The data indicate that melanoma cells exhibit differences in the adhesion properties under fluid shear and differences in the expression of adhesion components that correlate with their metastatic potential. PMID- 8713482 TI - Receptors for fibroblast growth factors. AB - The recent discovery of the involvement of heparan sulfate proteoglycans (HSPG) in the activation of fibroblast growth factor receptors (FGFR) has led to an intensification of study of this field. It appears that the HSPG act as low affinity receptors to which the fibroblast growth factors (FGF) must bind in order to successfully activate the high affinity FGFR. Heparan sulfate chains consisting of alternately arranged N-acetylated or N-sulfated glucosamine and uronic acid disaccharide regions, covalently attached to a core protein are found in two major families of cell surface HSPG, the syndecans and glypicans. A high affinity bFGF binding region has been isolated from fibroblast HS. There are four basic members of the FGFR family (FGFR 1-4), as well as a wealth of splice variants. The alternative forms of the basic receptors can have altered ligand binding or signalling qualities, depending on the region of the gene which is spliced. Investigations with null FGFR, incapable of signalling, have demonstrated the requirement for FGF in the organization of mammalian tissues and in embryonic patterning. Mutation of the FGFR genes has been recognized recently in human craniosynostoses where a single base pair mutation in the FGFR gene results in skeletal malformations specific to each syndrome. One suggestion is that the interaction of the mutant FGFR with the HSPG/FGF complex somehow contributes to the disease phenotype. PMID- 8713483 TI - Home intravenous anti-infective therapy (HIVAT): do the benefits outweigh the risks? AB - Home intravenous anti-infective therapy (HIVAT) has become an accepted mode of treatment worldwide. There are a number of risks intrinsic to such therapy which can be reduced by a team approach, coupled with careful patient selection criteria, a structured follow-up process and an emphasis on patient education. The benefits of HIVAT include the well-being accrued as a result of being at home, the possibility of returning to work or school, and a reduced risk of infectious and intravenous (IV) cannula-related problems. From a societal standpoint, economic cost-benefit analyses provide ample evidence of cost effectiveness. Studies indicate that potential risks, such as noncompliance, interruption of therapy, drug and IV catheter-associated problems, are infrequent. In the US, reimbursement for HIVAT services for the elderly remains a problem. PMID- 8713484 TI - HMG-CoA reductase inhibitors: issues in assessing their benefits in coronary heart disease. PMID- 8713486 TI - Cyproterone acetate: is it hepato- or genotoxic? AB - The preclinical safety assessment of cyproterone acetate (CPA) with regard to liver tumorigenesis was based on tumorigenicity studies, which revealed no mutagenic potential. Recently, in vitro studies on the formation of adducts and the enhancement of DNA repair synthesis with CPA have been published. These results are not unique to CPA, and the role of adducts and increased DNA synthesis in mutagenesis is still not clear. Dose-related hepatic toxicity has been reported with the prolonged use of CPA. An active surveillance study of patients taking long term CPA treatment has shown no correlation between the duration of CPA treatment and the prevalence of liver enzyme elevations. In a multicentre surveillance study of long term CPA use in 2506 patients included so far, not a single case of hepatocellular carcinoma has been observed. These findings do not support the theory of an elevated risk of hepatocellular carcinoma as a result of CPA treatment. In conclusion, there have been no observations which could point to an increased risk of proliferative liver change as a result of CPA treatment. PMID- 8713485 TI - Benefits and risks of HMG-CoA reductase inhibitors in the prevention of coronary heart disease: a reappraisal. AB - Although several cholesterol-lowering interventions have reduced coronary heart disease (CHD) events in clinical trials, drug therapy for hypercholesterolaemia has not been as widely used as the US and European guidelines recommend, mainly because until recently there was insufficient clinical trial evidence for improved survival. The Scandinavian Simvastatin Survival Study (4S) is the first trial of lipid-lowering therapy to demonstrate an unequivocal reduction in total mortality. Largely as a result of this study, there is now little disagreement on the necessity to reduce low density lipoprotein (LDL) cholesterol effectively in hypercholesterolaemic patients with CHD. Many physicians believe it is also important to reduce elevated levels of LDL cholesterol in patients without overt coronary disease, but more clinical trial evidence will be required before this is universally accepted. Inhibitors of HMG-CoA reductase are the most effective class of agents for this purpose, and have become widely used. It is likely that the magnitude of risk reduction produced by lipid-lowering therapy is proportional to the degree of cholesterol lowering achieved, which is an important consideration when selecting an agent and deciding the dosage to use. The results of several multicentre comparative trials have clearly established that the 4 members of the class are not all equipotent on a mg basis in terms of their effects on lowering LDL cholesterol. They have shown that the hypolipidaemic effect of simvastatin 5 mg approximately equals that of pravastatin 15 mg and lovastatin 15 mg and that of fluvastatin 40 mg, all given once daily. The tolerability profiles of HMG-CoA reductase inhibitors are excellent. Five-year data are available for simvastatin and lovastatin, and to date there is no good evidence for important differences in safety or tolerability among the class. PMID- 8713488 TI - Fetal abnormalities and use of substances sold in 'herbal remedies' shops. PMID- 8713489 TI - Hypothesis: an angiotensin converting enzyme genotype, present in one in three caucasians, is associated with increased mortality rate. AB - 1. This review argues that the deletion (D) allele of an insertion (I)/deletion polymorphism of the angiotensin I-converting enzyme (ACE) gene is a marker for a variant associated with increased ACE expression, as well as myocardial infarction (MI) and other life-threatening conditions. 2. By examination of I/D frequency in different age groups of individuals having well-known risk factors, it appears that homozygosity for the D allele may be associated with an increased risk of premature death in subjects at high-risk of cardiovascular events. For the risk factor hypertension, the odds ratio for DD vs II in patients aged > or = 60 years was 6.6. 3. Besides in MI itself, the DD genotype appears to be also more prevalent in MI patients who develop restenosis several months after balloon angioplasty, patients with various forms of heart failure, those with ventricular hypertrophy and diabetic patients who develop nephropathy. 4. Particular genotypes of other components of the renin-angiotensin system may add to the risk conferred by the ACE DD genotype. 5. Emerging evidence therefore suggests that the ACE genotype may eventually be placed on the list of common, well-known risk factors for fatal cardiovascular events. PMID- 8713490 TI - Urinary NO3- excretion as an indicator of nitric oxide formation in vivo during oral administration of L-arginine or L-name in rats. AB - 1. Endothelium-derived nitric oxide (NO), a major modulator of vascular tone, is synthesized from the terminal guanidino nitrogen of L-arginine. This reaction is inhibited by analogues of L-arginine, such as N-nitro-L-arginine methyl ester (L NAME). Many of the biological effects of NO are mediated by the second messenger cGMP. NO is rapidly oxidized to NO3-, which, like cGMP, is eliminated via excretion into the urine. In a placebo controlled study, we investigated whether oral bolus administration of L-arginine and L-NAME affects the urinary excretion rates of NO3- and cGMP in Munich Wistar Frommter (MWF) rats. 2. Twenty MWF rats were kept in metabolic cages and received L-arginine (3 g/kg bodyweight), L-NAME (50 mg/kg), or placebo (0.9% saline) in randomized order. Urine samples were sequentially collected for 10 h and analysed for creatinine, NO3- and cGMP. 3. L Arginine inducted a slight, but prolonged increase in urine flow, whereas L-NAME induced an early, transient increase in urine flow which was followed by a decrease. Creatinine clearance decreased by 65% after L-NAME, but was not affected by L-arginine or placebo. 4. Urinary NO3- and cGMP excretion rates transiently increased after L-arginine (NO3-: + 29%; cGMP: +16%) for 4-5 h, whereas L-NAME induced an immediate, pronounced and lasting inhibition of urinary NO3- and cGMP excretion (NO3-: -76%; cGMP: -46%). Urinary NO3- and cGMP excretions were significantly correlated (r = 0.755; P < 0.001). 5. Urinary excretion rates of NO3- and cGMP, expressed as mu mol/h, were correlated to urine flow (mL/h; r = 0.617 and 0.649, respectively; both P < 0.05), whereas after correction by urinary creatinine (mu mol/mmol creatinine) no correlation with urine flow was observed, indicating that these excretion rates were independent of renal excretory function. Thus we conclude that changes in the urinary excretion rates of NO3- and cGMP represent changes in NO production rates in vivo when expressed in relation to urinary creatinine. Urinary NO3- and cGMP excretion is modulated by acute NO synthase inhibition or substrate provision. PMID- 8713487 TI - Adverse drug events related to dosage forms and delivery systems. AB - While some of the adverse events caused by the administration of medicines are specifically attributable to the drug molecule, a proportion arises because of the chemical, biological and physical nature of the formulation. The effects may be compounded by certain patient factors, an incomplete understanding of the behaviour of the formulation or the coadministration of other drugs. This review examines adverse drug reactions and other adverse events arising from the nature of the dosage form or formulation used. These adverse effects may be the result of local irritation/toxicity, hypersensitivity or allergic reactions, systemic effects from essentially local therapies, or idiosyncratic reactions in a small number of individuals. In certain cases where the exact nature of the formulation is unknown, adverse events cannot be attributed to any single ingredient. In addition, the total of all ingredients of a formulation, even where details of the formulation are clear, may give rise to abnormal behaviour of the formulation in vivo. Often the desired objective of a particular specialised formulation leads to an unforseen but related adverse effect, and in certain instances these events are completely unpredictable and at variance with the perceived objectives of the formulation. PMID- 8713492 TI - Functional responses of the rat isolated seminal vesicle to electrical field stimulation: a pharmacological analysis. AB - 1. Electrical field stimulation (EFS) of the rat isolated seminal vesicle elicited frequency-dependent and tetrodotoxin sensitive contractions which were unaltered by hexamethonium or mecamylamine. 2. Prazosin alone was not sufficient to abolish these responses, but a combination of atropine and prazosin was fully effective, indicating involvement of both noradrenergic and cholinergic mechanisms. 3. Responses were predominantly cholinergic (blocked by atropine, potentiated by ecothiopate but not significantly altered by prazosin or guanethidine) at 1-8 Hz but became increasingly noradrenergic (blocked by prazosin or guanethidine but relatively unaltered by atropine or ecothiopate) with increasing frequencies of stimulation. 4. Electrical field stimulation of seminal vesicles removed from reserpine or 6-hydroxydopamine (6-OHDA)-pretreated rats produced contractions that were clearly cholinergic in nature. 5. After exposing the seminal vesicles to guanethidine, or after pretreatment of rats with 6-OHDA, responses to EFS remained, indicating that activation of discrete cholinergic and noradrenergic innervations seem to underlie the contractile responses observed. 6. Yohimbine and prazosin potentiated the predominantly cholinergic responses at 1, 2 and 4 Hz in tissues from untreated rats, but not in those from animals pretreated with reserpine or 6-OHDA, indicating the possibility of an interaction between the two innervations. 7. No inhibitory responses to EFS could be demonstrated in tissues precontracted with KCl in the presence of a combination of atropine and prazosin suggesting the absence of a nonadrenergic, noncholinergic inhibitory innervation. PMID- 8713491 TI - Selectivity of omega-conotoxin GVIA for n-type calcium channels in rat isolated small mesenteric arteries. AB - 1. The selectivity of omega-conotoxin GVIA (omega-CTX) for prejunctional N-type voltage-operated calcium channels (VOCC) was examined in rat isolated small mesenteric arteries mounted in a Mulvany-Halpern myograph. Contractile responses to perivascular nerve stimulation, noradrenaline (NA) and potassium (K+) were obtained before and after treatment with omega-CTX. The effects of omega-CTX were compared with those of felodipine, an L-type VOCC blocker. 2. Omega-CTX (3 nmol/L 10 mu mol/L) inhibited contractions to electrical field stimulation by up to 94%, compared with the corresponding time control group. Felodipine (0.1 mu mol/L) had little effect on the contractions to electrical stimulation compared with the vehicle-treated vessels. 3. Concentration-response curves to exogenous NA (0.1 mu mol/L) and contractions to a submaximal concentration of K+ (50 mmol/L) were unaffected by omega-CTX (3 nmol/L-10 mu mol/L). In contrast, the maximum contraction to NA in vessels exposed to felodipine (0.1 mu mol/L) was reduced by 37%, and the contraction to K+ (62 mmol/L) was reduced by 84% compared with vehicle-treated arteries. 4. The results indicate that even at concentrations up to 10 mu mol/L (10 000-fold higher than required to inhibit prejunctional N-type VOCC), omega-CTX inhibits only neurotransmitter release. Its effects are clearly different to felodipine as omega-CTX has no effect on post-junctional alpha1 adrenoceptor-mediated vasoconstriction or direct smooth muscle depolarization considered to be mediated by L-type VOCC. Therefore, at least at the vascular neuroeffector junction, omega-CTX appears to be highly selective for N-type VOCC with no effect on L-type VOCC. PMID- 8713493 TI - Endogenous nitric oxide production is augmented as the severity advances in patients with liver cirrhosis. AB - 1. Since endothelium-derived nitric oxide (NO) is a potent vasodilator and degraded into nitrous ions, we measured the serum nitrate ion (NO3-) and the amount of urinary excretions of NO3- as an index for endogenous NO to ascertain whether NO formation is augmented in patients with chronic liver diseases. 2. Using inpatients suffering from chronic liver diseases, serum levels and urinary excretions of NO3- were measured by using high-performance liquid chromatography with an anion exchange column. 3. Among the four patient groups of normal controls, and those with chronic liver diseases such as chronic active hepatitis, compensated cirrhosis, and decompensated cirrhosis the serum level of NO3- showed the highest level in a patient group with decompensated cirrhosis. The amount of urinary excretion of NO3- was significantly increased in both groups of patients with liver cirrhosis compared with the control group and patients with chronic active hepatitis. Patients with chronic active hepatitis did not show any difference between the normal control group. The amount of urinary excretion of NO3- correlated significantly and negatively with the level of serum albumin (P < 0.05) and counts of platelets (P < 0.01) in patients with compensated cirrhosis. 4. These findings suggest that the production of endogenous NO is augmented in patients with liver cirrhosis, particularly in a decompensated subgroup. Increases in the production of endogenous NO correspond to the progress of liver cirrhosis, but not in patients with chronic hepatitis. PMID- 8713494 TI - Comparison of effects of vasoactive agents on adenylate cyclase activity in endothelial and smooth muscle cells from the same specimen of fetal bovine aorta. AB - 1. Adenylate cyclase activity in vascular endothelial cells (EC) has not been adequately defined. We compared adenylate cyclase activities in EC and smooth muscle cells (SMC) from the same specimen of fetal bovine aorta. 2. The basal adenylate cyclase activities of EC and SMC did not differ significantly (18.9 +/- 0.8 and 21.4 +/- 1.7 pmol/mg protein per min, n = 10, respectively). 3. The adenylate cyclase of EC responded dramatically to catecholamines, with the ED50 value for isoproterenol being 0.036 mu mol/L, and was also more sensitive to calcitonin gene-related peptide than that of SMC. 4. The adenylate cyclase of SMC was more sensitive to prostaglandins (with the ED50 for PGI2 being 0.024 mu mol/L) and glucagon than that of EC, and responded modestly to catecholamines and was almost selectively sensitive to beta1-adrenoceptor agonists. 5. Maximum responses of adenylate cyclase to F-, guanosine 5'-o-(3-thiotriphosphate) and forskolin were greater in SMC than in EC. 6. Based on these findings, it was concluded that EC and SMC differed significantly in adenylate cyclase responsiveness to agonists including hormones, prostaglandins, peptides and substances which may modify the effects of G proteins, although they shared a common developmental origin. Presumably, beta-adrenoceptor agonists in EC and prostaglandins in SMC may appear to play an important role in cellular functions which are mediated by increases in cAMP. PMID- 8713495 TI - Effects of ACE inhibition and beta-blockade on collagen remodelling in the heart of Bio 14.6 hamsters. AB - 1. The effects of angiotensin converting enzyme (ACE) inhibition and beta blockade on collagen in the heart and on plasma catecholamines and tissue angiotensin (Ang) I and II were examined in Bio 14.6 Syrian hamsters. Male hamsters (76-79 days old) were given low-dose enalapril (3 mg/kg per day), high dose enalapril (30 mg/kg per day), atenolol (50 mg/kg per day) or vehicle for 65 days. Age and sex matched healthy F1b hamsters were used as controls. Collagen concentration was determined by measuring hydroxyproline content and the relative proportion of type I, III, and V collagens was obtained by non-interrupted sodium dodecyl polyacrylamide gel electrophoresis (SDS-PAGE). Per cent collagen area (PCA) was measured by pixel counting in myocardial tissue by a personal computer. 2. Although heartweight (HW) and bodyweight (BW) in F1b controls were significantly higher compared with drug-treated groups and vehicles, the HW/BW ratio in cardiomyopathic Bio 14.6 hamsters tended to be high compared with F1b controls and was decreased by each drug treatment. 3. Collagen concentration, total collagen content and PCA in the heart of Bio 14.6 hamsters were significantly higher than F1b controls. Collagen concentration and total collagen content were significantly decreased in all drug-treated groups compared with vehicles. 4. The proportion of type I collagen tended to decrease while that of type III collagen tended to increase in all drug-treated groups compared with vehicles. Type V collagen in vehicle-treated group was significantly higher than in F1b controls, while it tended to decrease in all drug-treated groups compared with vehicles. 5. Plasma concentrations of catecholamines (adrenaline and noradrenaline) were decreased significantly by atenolol and high-dose enalapril, but not by low-dose enalapril. Tissue AngI remained unaltered in any of the drug treated hamsters. Tissue AngII was decreased by the high-dose enalapril and beta blockade, and tended to be decreased by low-dose enalapril treatment. 6. These results reveal that enalapril and atenolol produced similar beneficial effects on collagen remodelling in Bio 14.6 hamsters by decreasing the total amount of collagen, and also by changing collagen phenotypes through the inhibition of the renin-angiotensin system. Both drugs also improved myocardial morphological integrity. PMID- 8713496 TI - N-nitro-L-arginine and indomethacin do not affect endothelin-induced constriction of large and small coronary arteries in the anaesthetized greyhound. AB - 1. The aim of this study was to investigate whether endothelin-1 (ET-1)-induced constriction of large and small coronary arteries in the anaesthetized greyhound is modulated by the endogenous release of nitric oxide or prostanoids. 2. ET-1 (1 100 ng/kg) and the alpha1-adrenoceptor agonist phenylephrine (0.5-2 mu g/kg), when injected directly into the circumflex coronary artery, caused dose-dependent decreases in epicardial coronary artery diameter and coronary vascular conductance without affecting systemic arterial pressure or the rate and force of cardiac contraction. 3. Inhibition of NO synthesis with N-nitro-L-arginine (NOLA, 5 mg/kg, i.c.) decreased coronary artery diameter, coronary conductance and heart rate and increased arterial pressure. The coronary vasoconstrictor response to ET 1 was unaffected by NOLA. By contrast, NOLA significantly increased the phenylephrine-induced constriction of the epicardial coronary artery but not the resistance vessels. 4. Indomethacin (5 mg/kg, i.v.), an inhibitor of cyclo oxygenase, significantly decreased epicardial coronary artery diameter but did not affect coronary conductance. Indomethacin had no effect on the coronary vascular responses to ET-1 or phenylephrine. Combined treatment with NOLA plus indomethacin also failed to affect the coronary vasoconstrictor effects of ET-1. 5. Basal release of NO and vasodilator prostanoids modulated resting coronary vascular tone but did not influence the vasoconstrictor responses to endothelin in either large or small coronary arteries. PMID- 8713497 TI - Alterations in fetal urine production during prolonged hypoxaemia induced by reduced uterine blood flow in sheep: mechanisms. AB - 1. Our aim was to identify mechanisms whereby prolonged fetal hypoxaemia alters renal function and urine production in fetal sheep. 2. Fetal hypoxaemia was induced for 24 h by reducing uterine blood flow at 129.0 +/- 2.1 days of gestation (term 145-147 days), causing a reduction in fetal arterial O2 saturation (SaO2) from 52.5 +/- 2.3 to 22.0 +/- 1.3% (P < 0.05). This hypoxaemia was initially associated with a mild acidaemia (pH 7.23 +/- 0.03). 3. The glomerular filtration rate (GFR) increased from a control value of 1.8 +/- 0.3 mL/min per kg to a maximal value of 2.8 +/- 0.6 mL/min per kg (P < 0.05) at 4-5 h of hypoxaemia, returning to control levels by 6-9 h of hypoxaemia. After 4 h of hypoxaemia renal blood flow was no different to control values (144 +/- 8 mL/min per 100 g kidney weight) but after 24 h of hypoxaemia it had increased to 190 +/- 8 mL/min per 100 g kidney weight (P < 0.05). Fractional reabsorption of Na+ in the proximal tubules decreased from a control value of 81.5 +/- 2.2 to 65.2 +/- 3.9% at 2-3 h of hypoxaemia (P < 0.05) and remained reduced (68.5 +/- 3.1%) at the end of hypoxaemia (P < 0.05). Fetal mean arterial pressure transiently increased (P < 0.05) but returned to control values by 4-5 h of hypoxaemia. Fetal renal vascular resistance was not significantly altered during hypoxaemia. Fetal urine production increased from a control value of 12.3 +/- 2.1 mL/h per kg to a maximal value of 19.1 +/- 4.2 mL/h per kg at 4-5 h of hypoxaemia (P < 0.05) and returned to control by 24 h of hypoxaemia. 4. Our results indicated that prolonged fetal hypoxaemia leads to the inhibition of Na+ reabsorption in the proximal portion of the renal tubules. Changes in GFR induced by hypoxaemia were similar to those in fetal urine production and were not associated with changes in renal blood flow. We conclude that prolonged fetal hypoxaemia affects renal haemodynamics and the reabsorptive capacity of the renal tubules, resulting in a diuresis. PMID- 8713498 TI - Calcium and calcium-related signalling pathways in glomerular mesangial cells. AB - 1. Calcium is an important cation for many of the physiological and pathophysiological functions of the mesangial cell. Calcium binds to many proteins in the cell. some of these proteins seemingly serve, primarily, as intracellular buffers for calcium, whose intracellular concentration must be strictly controlled. In addition to these buffering proteins, a large number of proteins are regulated by calcium. These proteins are important for mesangial cell structural integrity, maintenance of internal ionic composition, contractility, effector responses to a variety of hormones and growth factors, and production of inflammatory mediators. 2. Calcium homeostasis in mesangial cells is complex. Cells respond to vasoactive peptides and growth factors with increases in intracellular free calcium ([Ca2+]i) due to Ca2+ release from intracellular storage sites and entry across the cell plasma membrane. 3. Individual mesangial cells respond to arginine vasopressin with repetitive [Ca2+]i spikes. Increasing the concentration of vasopressin up to 10 nmol/L increases the frequency of the repetitive Ca2+ spikes. The amplitude of the oscillations does not vary with the concentration of vasopressin used. 4. The presence of Ca2+ oscillations whose frequency varies with ligand concentration suggests that the Ca2+ signal may encode its information through a frequency dependent mode in addition to, or rather than, an amplitude-dependent mode. Calcium can change the conformation of proteins and can change catalytic activity of enzymes directly. 5. Another important mode of regulation by calcium is demonstrated by cytosolic phospholipase A2, where calcium is important not for catalytic activity directly but for the apposition of enzyme with substrate so that the catalytic activity may be manifest. PMID- 8713499 TI - Regulation of rat kidney mesangial cell phospholipase A2. AB - 1. The precursor of eicosanoids is arachidonic acid, which emanates from the cleavage of the sn-2 position of phospholipids by phospholipase A2 (PLA2). Eicosanoids have diverse physiological and pathophysiological effects in the kidney. The regulation of phospholipase A2 has important implications for kidney function. 2. In the current communication we focus our attention on mesangial cell cytosolic PLA2 (cPLA2) and its regulation at the post-translational and post transcriptional level. 3. At the post-translational level, using site directed mutagenesis of cPLA2 and a dominant negative ras, we have demonstrated that cPLA2 can be phosphorylated by mitogen activated protein (MAP-2) kinase leading to increased cPLA2 enzymatic activity. 4. At the post-transcriptional level we show that the half-life of cPLA2 mRNA in mesangial cells is significantly increased when mesangial cells are stimulated by mitogens. We further demonstrate the presence of three ATTTA motifs in the 3' untranslated region (3' UTR) of the cPLA2 cDNA. 5. Using chimeric constructs bearing the 3' UTR from rat cPLA2 fused downstream of the luciferase reporter, we demonstrate that this region exerts a destabilizing effect on cPLA2. 6. We have isolated and mapped genomic DNA and polymorphic markers for cPLA2 in the human and rat. PMID- 8713500 TI - Physiological role of large, Ca2+-activated K+ channels in human glomerular mesangial cells. AB - 1. Contraction assays and patch clamp methods were used to determine the role of K+ channels in the regulation of contractile tone of human mesangial cells (MC) in culture. 2. MC contraction was induced by vasoconstrictor agents, such as angiotensin II (AngII; 100 nmol/L) and glybenclamide (Glyb), but not by iberiotoxin (IbTX), a blocker of large Ca2+-activated K+ channels (BK(Ca)). These results suggest that Glyb-sensitive K+ channels, but not BK(Ca) channels, were active at rest. 3. In the presence of 100 nmol/L IbTX, contraction by AngII was slightly, but not significantly, enhanced, indicating that BK(Ca) has a minimal role as a negative feedback regulator of contraction. Nitroprusside (NP; 100 mu mol/L) a nitric oxide (NO) donor, atrial natriuretic peptide (ANP; 1.0 mu mol/L) and db-cGMP (10 mu mol/L) attenuated AngII-induced contraction in the absence, but not in the presence, of IbTX, suggesting that BK(Ca) channels were activated by cGMP. 4. In patch clamp experiments, three distinct K+-selective channels of 9, 65 and 150 pS (outward currents) were found in excised, inside-out patches. The 150 pS channel was completely inhibited by 100 nmol/L IbTX and displayed voltage- and calcium-dependent gating qualitatively similar to BK(Ca) in other cell types. 5. In cell attached (CA) patches, the response of BK(Ca) to bath AngII (100 nmol/L) was relatively minor in control solutions, but was considerably greater in the presence of db-cGMP. 6. In excised patches, Mg-ATP (1 mmol/L) plus db-cGMP (1 mu mol/L) activated BK(Ca) in the absence, but not the presence, of the non-specific kinase inhibitor, staurosporine. 7. Separate experiments showed that BK(Ca) were also activated by arachidonic acid and high ambient glucose concentrations. 8. These results indicate that: (i) resting MC tone is sensitive to glybenclamide and apamin; and (ii) the role of BK(Ca) as a negative feedback regulator of contraction is minimal under normal conditions but is markedly enhanced by cGMP-stimulating relaxants and arachidonic acid. PMID- 8713501 TI - Angiotensin II-induced tyrosine phosphorylation in mesangial and vascular smooth muscle cells. AB - 1. Angiotensin II (AngII)-induced, activation of phospholipase C (PLC) and Ca2+ dependent Cl- channels is an important signal transduction pathway for the regulation of vascular smooth muscle cell (VSMC) and glomerular mesangial cell contraction and growth. While AT receptors are traditionally thought to be G protein coupled to the beta isoform of PLC, recent evidence suggests that in some tissues AT receptors may also activate the PLC-gamma isoform via tyrosine phosphorylation. 2. By western analysis, we identified PLC-gamma1 in the above cell types. We found that within 3 min of exposure to 10(-7) mol/L AngII, tyrosine phosphorylation of PLC-gamma1 was observed; however, peak response (>3 fold increase) occurred within 0.5 min. In addition, pre-incubation of these cells with the tyrosine kinase inhibitor genistein blocked the tyrosine phosphorylation of PLC-gamma1 by AngII. In contrast, preincubation with the tyrosine phosphatase inhibitor sodium vanadate increased the levels of tyrosine phosphorylation of PLC-gamma1. Similar results were found when intracellular levels of 1,4,5-IP3 were measured after AngII exposure. 3. By using patch clamp techniques on cultured rat mesangial cells exposed to AngII, we found that the release of 1,4,5-IP3-sensitive intracellular Ca2+ stores stimulated low conductance Cl- channels. Preincubation with genistein, abolished the usual 10 fold increase in Cl- channel activity observed with AngII. 4. Therefore, we conclude that in VSMC and glomerular mesangial cells (i) AngII transiently stimulates PLC activity via tyrosine phosphorylation of the gamma1 isoenzyme, (ii) tyrosine phosphorylation of PLC-gamma1 and production of 1,4,5-IP3 in response to AngII is dramatically inhibited by tyrosine kinase inhibition and stimulated by tyrosine phosphatase inhibition, (iii) activation of Ca2+-dependent Cl- channels by AngII-induced release of 1,4,5-IP3-dependent intracellular Ca2+ stores is also abolished by tyrosine kinase inhibition. In summary, this AngII induced signal transduction cascade provides a possible mechanism for both the contractile and growth-stimulating effects of AngII on VSMC and glomerular mesangial cells. PMID- 8713502 TI - Regulation of mesangial chloride channels by insulin and glucose: role in diabetic nephropathy. AB - 1. In response to vasoactive peptides (e.g. angiotensin II (AngII), vasopressin, endothelin-1, platelet-activating factor), glomerular mesangial cell contraction is mediated through activation of a Ca2+-dependent Cl- conductance that, in turn, promotes membrane depolarization and voltage-activated Ca2+ entry. 2. Using patch clamp technology, our laboratory was the first to characterize a candidate Ca2+ dependent, 4 pS Cl- channel that is stimulated by vasoactive peptides in cultured rat mesangial cells. In the absence of extracellular insulin, the activation of Cl- channels by AngII is abolished. We find that Cl- channel sensitivity to intracellular Ca2+ and the membrane density of AngII receptors is also dependent on the presence of insulin. 3. Our studies also show that high extracellular glucose interferes with mesangial cell IP3 generation and Cl- channel stimulation. Importantly, we find that the insulin-dependency of Cl- channels occurs within the range of plasma insulin concentrations observed in normal, obese, hypertensive and diabetic humans (i.e. 1-100 mu U/mL). Similarly, normal regulation of Cl- channel activity is also modulated by glucose concentrations commonly observed in the plasma of diabetic humans (5-30 mmol/L). 4. There is substantial evidence, both in diabetic humans and animal models, that the provision of insulin and improved glycaemic control corrects or prevents glomerular hyperfiltration. The requirement for normal insulin and glucose levels, for the proper regulation of the 4 pS Cl- channel, provides a mechanism for impaired Ca2+ uptake and contraction observed in glomerular mesangial cells in association with insulin deficiency and hyperglocaemia. PMID- 8713503 TI - Increasing resting vascular tone in conscious dogs does not alter the mesenteric vasoconstrictor responses to ANP. AB - 1. There is considerable in vitro evidence that, at high concentrations, atrial natriuretic peptide (ANP) acts directly on pre-constricted blood vessels to cause vasorelaxation. Previously, we have seen vasoconstriction rather than vasodilatation in conscious dogs at physiological levels of the peptide. It is possible that the low resting vascular tone in our conscious, unstressed animals prevented the manifestation of the relaxant properties of ANP in vivo. 2. In the present study in conscious, instrumented dogs, we studied the mesenteric vascular responses to 10 min infusions of ANP (10, 25, 50 and 100 ng/kg per min, i.v.) when resting vascular tone was enhanced with a continuous infusion of AVP (75 pg/kg per min, i.v.) and compared these with responses in the normal condition (no added AVP). 3. Mesenteric vascular resistance was increased by ANP (10, 25, 50 and 100 ng/kg per min) by 9 +/- 2, 20 +/- 6, 29 +/- 7 and 32 +/- 9%, respectively. Increased resting vascular tone did not alter the mesenteric vasoconstrictor response to ANP. Thus, the discrepancy between in vitro (vasorelaxation) and in vivo (vasoconstriction) findings may be the result of the widely different concentrations of ANP used, rather than the state of resting vascular tone. PMID- 8713504 TI - [Nutrition and bone mineral density in premenopausal women]. AB - Environmental factors have an important role in osteoporosis. Diet and, in particular, nutrients like calcium, vitamin D or phosphorus were extensively studied as determinants of bone mineral density, but the results remain conflicting and there is no clear evidence for an independent effect of such factors in the bone density of premenopausal women. We studied 66 healthy premenopausal women (20-40 years-old) aiming to relate bone mineral density, as measured in three different sites (distal forearm, lumbar spine and femoral neck) using single X ray and dual energy X-ray absorptiometry, with nutritional intake as estimated by a semi-quantitative food frequency questionnaire. Demographic, anthropometric and other life style variables were also assessed. There was a significant correlation between distal forearm and femoral neck (r = 0.57) or lumbar spine (r = 0.45) bone mineral density. No significant effect of age was observed for distal forearm bone mineral density in these women. In a stepwise multiple linear regression model, evaluating the contribution of all the variables studied, only body mass index (p=0.038) and vitamin A ingestion (p = 0.020) had an independent contribution for the variation in distal forearm bone mineral density. Mean bone mineral density, assessed in the femoral neck (p = 0.003) or the lumbar spine (p = 0.056) was different across tertiles of alcohol ingestion, being higher in non-drinkers. However, among regular drinkers there was a significant positive correlation between alcohol ingestion and femoral neck bone mineral density (Spearman's r = 0.53, p = 0.015). This study shows that the effect of nutrition seems dependent on the anatomical site assessed and that there is a weak correlation between nutritional intake and the actual bone mineral density. PMID- 8713505 TI - [Quality control of the micro-ELISA technic applied to the diagnosis of human and canine visceral leishmaniasis]. AB - Visceral leishmaniasis (VL) or kala-azar is, in Portugal, a zoonosis with the dog as reservoir. A quality control of the technique of micro-ELISA was carried out, using as reference the technique of IFI, the most commonly used for the diagnosis of this protozoosis, both for human and canine sera. Three different methods were used to estimate the cut-off point: X + 2sd (average for negative sera plus two standard deviation), P/N and J index. As quality parameteres were used sensitivity, specificity, efficacy and positive predictive value. The cut-off point for human sera was established at 0.100 A, with 100% sensitivity, 90.5% specificity, 95.3% efficacy and 91.4% positive predictive value, and for canine sera in 0.200 A, with 80.0% sensitivity, 94.3% specificity, 87.7% efficacy and 96.6% positive predictive value. Reproducibility was not fully satisfactory and two different ways of improving it are proposed: P/N and a correction factor. A statistically significant correlation was observed between micro-ELISA's absorbances and IFI titres regarding human sera, though it was not possible to do the same for dog sera. PMID- 8713506 TI - [Testicular relapse in acute lymphoblastic leukemia. The experience of a pediatrics service]. AB - Testicular relapse in Acute Lymphoblastic Leukemia is a problem which has been developing more and more, as the therapeutic protocols of acute lymphoblastic leukemia are improving. Its appearance, specially during chemotherapy, makes the prognosis of the disease poorer. There have been some diagnostic methods, used in the screening of this situation. The use of more aggressive therapeutic protocols can lead to more encouraging results. The authors of the research present seven children with testicular relapse, which represent 17% of the total group of fourty one boys with acute lymphoblastic leukemia, treated at St. o Antonio Pediatric Ward. They analyse the time of onset, the diagnostic approach of the testicular lesion, the therapeutic protocols chosen for each case and the results. PMID- 8713507 TI - [Total cholesterol and high-density lipoprotein cholesterol in patients with non insulin-dependent diabetes mellitus]. AB - Non-insulin-dependent diabetics often have quantitative changes in plasma lipid profiles characterised by higher triglycerides and lower HDL-cholesterol than the average population. This paper summarises the cross-sectional data (reported by the general practitioners participating in Medicos-Sentinela) concerning total and HDL-cholesterol in a cohort of non-insulin-dependent diabetics treated at primary care settings in Portugal. Total cholesterol and High Density Lipoprotein (HDL) associated cholesterol were significantly higher in women. Total cholesterol increased significantly with age (in women), regular alcohol intake, body mass index, systolic blood pressure and diastolic blood pressure (in males). HDL-cholesterol showed significant increase with age (both sexes and males only), gender, and alcohol intake in males. The increase in total cholesterol found in patients with regular alcohol intake is an infrequently reported finding. PMID- 8713508 TI - [Frontal lobe-type dementia associated with motor neuron disease]. AB - A case of presenile dementia with dominant frontal disfunction, progressive aphasia and Motor Neuron Disease with prominent bulbar signs is reported. Considering the clinical examination, the measurements of the regional cerebral flow (SPECT) and the histological appearances, we suggest the diagnosis of Dementia of Frontal Lobe Type and Motor Neuron Disease. We reviewed other disorders labelled Primary Frontal or Fronto-temporal Dementias and we discuss this new dementia and the difficulty in its classification. PMID- 8713509 TI - [Intestinal endometriosis]. AB - Pelvic endometriosis affects 5 to 10% of all menstruating women, and of these 3% to 34% have intestinal involvement. Commonly, intestinal endometrial lesion affects only the serosa. Although the invading endometrial tissue may extend into the intestinal wall, it does not usually reach the mucosa. The majority of the patients are asymptomatic. The most common symptom is lower recurrent abdominal pain. It has been estimated that 0,9% of women with intestinal endometriosis have obstructive symptoms. Occasionally deeper and more extensive intestinal wall involvement results in cyclic menstrual rectal bleeding. The authors describe a case with obstructive symptoms associated to rectal bleeding. This is an uncommon finding. Histological confirmation of deep muscular and mucosal involvement was obtained. This case led us to some considerations about the diagnostic difficulties and therapeutic possibilities in the present. PMID- 8713510 TI - [A diaphragmatic pacemaker in the treatment of ventilatory failure]. AB - Diaphragmatic Pacing was used to treat ventilatory failure in a young boy with bilateral diaphragmatic palsy secondary to cervical cord injury, on chronic mechanical ventilation for 6 months. The indications for diaphragmatic pacing in ventilatory failure were discussed, as well as the assessment of phrenic nerve integrity and the pacing schedules utilized to slowly improve the diaphragm endurance and achieve a full-time weaning from mechanical ventilation. PMID- 8713512 TI - [How to make a surgical curriculum]. AB - Discussion on curriculum vitae is a highly valorized parameter in the final exam to obtain the degree of medical specialist in Portugal. However, the lack of guidelines for making up a curriculum causes difficulties for everyone involved: residents, juries and institutions which govern the equity of the process. This paper presents guidelines for a curriculum which can be applied to any surgical speciality. The proposal is based on the regulations which govern post-graduate medical training. A broad debate on this issue is also suggested in this article. PMID- 8713511 TI - [Spinal epidural empyemas]. AB - We report ten patients harbouring spinal epidural abscess, aiming to evaluate the factors that may lead to an early diagnosis and that can eventually influence the prognosis. There were seven males and 3 females, with ages comprised between 17 and 66 years. Abscesses were localised mainly in the dorsal region. The most important predisposing factors were infections or other disorders know to be related with compromise of the immunological system. Back pain with or without signs of spinal cord involvement was the most frequent clinical presentation. The pretreatment average time was 16,3 days. Staphylococcus aureus was the most commun organism isolated. Erythrocyte sedimentation rate (ERS) was uniformly elevated, being the most important laboratory data for the diagnosis of this situation. Diagnosis was frequently made on clinical grounds but it was always confirmed by myelography with computed tomography or magnetic resonance. All patients were submitted to surgical drainage. Two patients recovered totally, 5 partially, 2 did not recover at all and 1 died. We conclude that the prognosis is related to the surgical delay and that it depends on the identification of the predisposing factors and the recognition of a clinical picture of spinal cord involvement associated to an elevated ESR. Magnetic resonance is the most reliable imaging technique. PMID- 8713514 TI - Emerging and re-emerging microbial threats to human health. PMID- 8713513 TI - [AIDS: the view from Africa]. AB - Epidemiological data on AIDS in several zones in Africa is presented. The clinical and diagnostic aspects of HIV infection in Africa, based on the definition of Bangui's AIDS (1986), are analysed and the criteria is presented. The treatment and follow-up of a patient with infusion by HIV in the African context is discussed. Reference is then made to some data on the status of AIDS in Angola. PMID- 8713515 TI - Systematic reviews and the Cochrane collaboration. PMID- 8713517 TI - Upper urinary tract stone analysis using X-ray diffraction: results from a tertiary referral centre in northern India. AB - BACKGROUND: The spectrum of urinary stone disease has changed considerably in India from the common childhood bladder stone to the more frequent upper tract calculi. We analysed the gravel retrieved from the upper urinary tract using X ray diffraction analysis in an attempt to evaluate the composition of the stones. METHODS: We analysed 434 upper urinary tract calculi from May 1993 to June 1994 obtained endourologically, as well as by extracorporeal shock wave lithotripsy and open surgery. The stones were analysed using a Phillips compact X-ray diffractometer (PW1840). The PC-APD software was used for data collection and peak search. The phase matching was done by the software using the JCPDS reference database. RESULTS: Oxalate stones comprised 97% of the total stones with calcium oxalate monohydrate forming 90% and calcium oxalate dihydrate and mixed stones forming the remainder. Struvite stones were found in 1.4%, while uric acid and apatite stones were less than 1%. There were no cystine calculi. Seventy per cent of calcium oxalate monohydrate and 40% of calcium oxalate dihydrate stones were pure. All the struvite and apatite calculi were almost pure. Only 15% of staghorns did not consist of oxalate. Nine of the ten stones in children were of the calcium oxalate monohydrate variety. The stone composition in females was similar to that in males. CONCLUSIONS: X-ray diffraction data indicate that urinary stone disease in north India is different from that in the western world. Calcium oxalate monohydrate stones predominate. These stones are hard to break and have a different metabolic origin from those consisting of calcium oxalate dihydrate. These findings might help in selecting the most appropriate method of treatment in north India and they indicate directions in which further metabolic studies might be planned. PMID- 8713516 TI - Long term morbidity in survivors of the 1984 Bhopal gas leak. AB - BACKGROUND: The extent and nature of long term health sequelae among survivors of the Bhopal gas disaster are not known. In 1994 an International Medical Commission was set up with the aim of assessing respiratory, neurological and other health effects attributable to gas exposure. METHODS: An epidemiological survey of a representative sample of gas-exposed inhabitants of Bhopal was conducted in January 1994; for reference, a group of unexposed persons in the same city were surveyed. Questionnaires regarding health and exposure were administered to 474 persons, and a random sample (n=76) were subjected to respiratory and neurological testing. Responses to the questionnaire and the results of clinical testing were analysed according to a measure of individual gas exposure. RESULTS: A large number of subjects reported general health problems (exposed v. unexposed; 94% v. 52%) and episodes of fever (7.5/year v. 2.5/year); adverse outcome of pregnancy (e.g. still-births, 9% v. 4%) and respiratory symptoms (81% v. 38%), with a strong gradient by exposure category. This was not accounted for by differences in smoking, and was consistent with the results of spirometric testing. Neurological and psychiatric symptoms were reported more frequently by subjects in high exposure categories and the results of neurological examination and testing tended to confirm this finding. Ophthalmic symptoms demonstrated a similar pattern. Although a number of other symptoms were reported (with the possible exception of gastrointestinal disease), there was no clear evidence of other organ system damage attributable to gas exposure. CONCLUSION: The gradient of reported symptoms and clinical test results with estimates of exposure among these survivors of the gas leak suggests that a proportion of their current respiratory and neurological disease was due to gas exposure. PMID- 8713518 TI - Correlation of computed tomography with second-look laparotomy in ovarian carcinoma. AB - BACKGROUND: Cancer of the ovary is the second commonest gynaecological malignancy after cancer of the cervix. Surgery followed by cisplatin-based chemotherapy is the standard treatment approach. In patients with persistent disease, second-look laparotomy offers an opportunity to debulk the tumour. This is usually followed by an alternative method of chemotherapy. We compared the findings at surgery (second-look laparotomy) with the preoperative computed axial tomography scan assessments. METHODS: Thirty-seven patients with epithelial ovarian carcinoma were assessed with computed axial tomography scans of the abdomen and pelvis prior to undergoing a second-look laparotomy. RESULTS: Tumour was correctly identified on computed axial tomography scan in 11 patients who had macroscopic evidence of cancer at laparotomy. In 6 patients, both computed axial tomography scan and surgery showed no disease recurrence. In the remaining 20 patients, there was a mismatch between the computed axial tomography scan and the surgical findings. In 16 of the 20 (80%) patients, computed axial tomography scans were negative but tumour was present. When the tumour was less than 1.5 cm in diameter it was missed in 8, and when equal to or greater than 1.5 cm, it was missed in 5 patients. These small tumour deposits were located in the retroperitoneal area, under the dome of the diaphragm, omentum or peritoneum, liver surface, and in the pouch of Douglas. In one case each, infiltration of the urinary bladder, sigmoid colon and rectum was also not detected. In 4 patients, computed axial tomography scans showed tumour when none was present. CONCLUSION: Computed axial tomography scan cannot detect small nodules often present in ovarian cancer, and thus even if a computed axial tomography scan is normal it should not exclude a second-look laparotomy. PMID- 8713519 TI - Cryptosporidium, isospora and strongyloides in AIDS. AB - BACKGROUND: Diarrhoea is a common presentation in patients with AIDS. It occurs due to a number of parasites which are seldom seen in immunocompetent hosts. METHODS: Between January 1993 and July 1994, faecal specimens from 77 patients with AIDS presenting with diarrhoea of more than a month's duration were examined by direct wet mount microscopy of saline and iodine preparations and by the modified Ziehl-Neelsen stain. RESULTS: Cryptosporidium, Isospora and Strongyloides stercoralis alone or in combination were present in 29 of the 77 patients. CONCLUSION: The presence of these parasites highlights their pathogenic potential for immunocompromised patients with AIDS. Diarrhoea due to Isosporais responsive to treatment, hence their identification has therapeutic implications as well. PMID- 8713520 TI - Dengue haemorrhagic fever and the dengue shock syndrome in India. AB - The clinical spectrum of dengue fever ranges from asymptomatic infection through severe haemorrhage and sudden fatal shock. Increased capillary permeability is the diagnostic feature of dengue haemorrhagic fever (DHF). The pathophysiology of DHF/dengue shock syndrome (DSS) is related to sequential infection with different serotypes of the virus, variations in virus virulence, interaction of the virus with environmental or host factors and a combination of various risk factors. Infection due to low virulence strains is assumed to be the reason for the infrequent incidence of serious dengue disease in India. Since all four serotypes of the dengue virus have been implicated in various outbreaks in this country and several outbreaks of DHF/DSS have been recorded since the first report in 1963, further epidemics of the disease are likely. The situation is aggravated by the recent emergence of DHF/DSS in Sri Lanka. In view of the potential of this disease to spread, effective preventive and control measures should be a priority. PMID- 8713521 TI - Chemotherapy for advanced non-small cell lung cancer. AB - Less than 20% of all cases of non-small cell lung cancer are operable. The treatment option available for such patients is either radiotherapy or chemotherapy. Various combinations of chemotherapeutic agents have been tried. The combination of cisplatin and 5-fluorouracil (5-FU) has been proved to have a synergistic antitumour effect in many experimental and clinical studies. In this paper the authors report the results of a trial using tegafur (which is a prodrug of 5-FU; 1-[2-tetrahydrofuryl]-5-FU) and uracil along with cisplatin in inoperable non-small cell lung cancer. Thirty-one patients were entered into the study, all of whom were less than 75 years old (mean age 61 years). The patients (except for 2) had either stage IIIB (12) or stage IV (17) disease with adenocarcinoma and squamous cell carcinoma in equal proportions (15 each). A combination of uracil and tegafur (400 mg/m2) in 100 mg capsules (100 mg tegafur and 224 mg of uracil), was given orally for 21 days. Most of the patients received 300 mg of the combination tablets twice a day. Cisplatin (80 mg/m2) was given as an infusion over 90 minutes on day 8 after adequately hydrating the patients. This cycle was repeated every 4 weeks. At least 2 treatment cycles were given, unless there was disease progression or toxicity. The response and survival rates were assessed after a follow up period which ranged between 9 and 30 months. The patients received 1 to 4 cycles of therapy and the response rates were: complete response--nil,partial response--11, no change--11 and disease progression--9. The overall response rate was 35% (95% CI: range 19-52%). Five patients achieved at least 50% tumour reduction after 2 cycles. Of the 11 patients who showed partial response, 5 (35%) had stage IV disease and 6 (36%) had other stages; 6 (40%) had squamous cell carcinomas and 5 (31%) had tumours of other histological types. The median duration of response was 6 months (3-13 months). In stage III disease, the 1-year survival rate was 31%, with a median survival time of 11 months, while in stage IV disease it was 29% with a median survival time of 8 months. There was a low incidence of toxicity, with anaemia (10%), leukopenia (6%) and thrombocytopenia (6%) being the most common side effects. There were also no treatment-related deaths. The authors concluded that oral uracil and tegafur were as effective as other combinations with cisplatin. They also caused very few side-effects. They suggest that a larger trial needs to be carried out. PMID- 8713522 TI - Spinal tuberculosis with neurological deficits. AB - Two hundred patients suffering from tuberculosis (TB) of the spine with neurological complications were the subjects of this review. They were graded according to the Frankel system into--A: complete neurological deficit; B: sparing of some sensation; C: sparing of sensation but no useful motor function; D: sparing of sensation and useful motor function; and E: no deficits. Investigations carried out included detailed neurological assessment, radiography, contrast myelography and, in the later stages of the study, spinal computerized tomography (CT) scan. The authors believe that contrast myelography provides the best indication of spinal compression in TB spine. Treatment was by surgical decompression followed by chemotherapy. The surgical approach for thoracic spine disease was by the anterior transthoracic route. Cervical lesions were also approached by the anterior route, lateral to the carotid vessels. Unexpected findings during operation included lymphoma, plasmacytoma, non tuberculous granulation tissue, salmonella osteomyelitis and tumour metastasis. Ten patients died during the postoperative period, all of whom had extensive systemic TB. When indicated, antituberculous drugs were administered postoperatively for two years and immobilization was done in a plaster cast for 3 months. Only 30 patients showed partial recovery. Improvement was found to be related to the grade of deficit; thoracic lesions with severe neurological deficits showed the least improvement while lumbar disease had the best outcome. The study recommended a combination of surgery and chemotherapy for all cases of TB spine with neurological deficits after a short delay during which respiratory function should be improved. The surgery should aim at decompression of the cord by removal of pus, granulation tissue and sequestra, with internal splintage with bone grafts to reduce the hospital stay. Conservative treatment is unwise because it is not always possible to distinguish between TB and neoplastic lesions. PMID- 8713523 TI - Pyrexia of unknown origin. PMID- 8713524 TI - Ethical issues in perinatology. PMID- 8713525 TI - Letter from Johannesburg. PMID- 8713526 TI - Letter from London. PMID- 8713527 TI - Letter from North America. Medical practice and criminal law. PMID- 8713528 TI - Call for specialization in health journalism, New Delhi, 28 August 1995. PMID- 8713529 TI - Medical College, Baroda. PMID- 8713530 TI - Too many people. PMID- 8713531 TI - Mushrooming nursing homes. PMID- 8713532 TI - Reply to Dr. Ritu Priya. PMID- 8713533 TI - Medical council of India. PMID- 8713534 TI - Are clinical skills redundant? PMID- 8713535 TI - Chronic ulcerative gingivo-stomatitis due to Corynebacterium ulcerance. PMID- 8713536 TI - A comparison of radiographic and anatomical evidence of tooth development in infant apes. AB - This study assesses dental development in a sample of 8 known-age infant orangutans and chimpanzees in order to examine discrepancies between radiographic analyses of tooth development and the actual developing teeth. All crania were radiographed and any developing teeth then extracted for comparison. The height of deciduous teeth was measured both on the radiographs and on the extracted teeth to assess the accuracy of radiographic tooth measurements. A comparison between the radiographic and actual measurements of the deciduous teeth reveals substantial discrepancies. For permanent teeth, the degree of radiographic underestimation of tooth development was found to vary depending on the tooth being studied, its stage of development (early stages are particularly problematic) and the radiographic techniques used. Examination of the dissected teeth also revealed earlier times of onset of calcification for I2(2), C1(1), P3 4(3-4) and M2 than reported in previous studies. For the canine and M2(2), upper and lower homologues appear to begin development at different times, but there is no apparent difference in onset times between other upper and lower homologues. PMID- 8713537 TI - Interpositions in conflicts between males in bimale groups of mountain gorillas. AB - In bimale groups of mountain gorillas, conflicts between the silverbacks sometimes induce infants and females to interpose between the opponents. This paper describes a set of 40 such interpositions, using data from 380 h of focal observation of the silverbacks in 2 groups at the Karisoke Research Center. Aggressive interactions between males can be disruptive for females and infants: they might prevent them from feeding and resting normally. Interpositions occurred in 10-25% of the conflicts between silverbacks. None of the males seemed to receive a greater proportion of interpositions. Interpositions increased the interval between conflicts in one of the research groups. As a mechanism for resolving conflicts, interpositions could be advantageous when control interventions are not possible, i.e. when the individuals in conflict are the most dominant of the group. Because they might reduce the rate of aggression between males, these interpositions could also be one of the mechanisms allowing male coexistence in gorillas. PMID- 8713538 TI - Polygyny in a free-ranging group of buffy-tufted-ear marmosets, Callithrix aurita. PMID- 8713539 TI - Grooming and social rank by birth: the case of Macaca fascicularis. PMID- 8713540 TI - Observations on task-related behaviour of Macaca arctoides. PMID- 8713541 TI - 'Peat-bathing' by captive white-faced capuchin monkeys (Cebus capucinus). PMID- 8713542 TI - A note on scavenging by wild chimpanzees. PMID- 8713543 TI - Benign prostatic hyperplasia in the nonhuman primate Leontopithecus. PMID- 8713544 TI - Cytogenetic study in a specimen of Cebus apella with an atypical phenotype. PMID- 8713545 TI - Planning, preparation, execution, and imagery of volitional action. AB - There are different motor sets, which a human subject can be in or act from: he or she can be in a self-initiated voluntary movement set (action) or in a response set (re-action). Also, imagery sets are available that are necessary for the acquisition and practice of skill. Most important are such imagery sets for rehearsal in theatre, dance, music, sports, combat, etc. PMID- 8713546 TI - On knowing how to do things: a theory of motor imagery. AB - The distinction between 'knowing how' and 'knowing that' is fundamental to current theories of cognition. Two distinct encodings or representations are implied, one conscious and verbalisable and the other normally unconscious yet demonstrable in behaviour. The paper discusses the nature of these two kinds of representation and relations between them. It is shown that imagery forms an essential mediating link between the two encodings and a theoretical model-the Action-Language-Imagination or ALI model-is presented. An important feature of the model is the role attributed to the motor system in generating imagery and principal features of motor imagery are reviewed in the context of the ALI model and with reference to recent experimental findings. Problems in mapping conscious representations of action onto physical brain mechanisms are briefly discussed. It is proposed that the physical basis of imaginal representations of actions is best understood in terms of the mechanisms of motor control. A two stage theory of motor imagery is proposed in which the first stage, the generation of a prototypical action is virtually identical to that involved in overt actions whilst the second stage depends on the retrieval of sensory impressions from memory. PMID- 8713547 TI - The bailiwick of visuo-spatial working memory: evidence from unilateral spatial neglect. AB - An accurate representation of the visual environment is crucial for successful interaction with objects in that environment. The means by which that representation is formed in working memory is a major focus of the issues and research discussed in this paper. The discussion draws on experimental studies of healthy adults and of patients with impairments of visual perceptual processing or of visuo-spatial mental representation. These disorders are most commonly linked with the disorder referred to clinically as unilateral spatial neglect. We discuss the observed dissociation between perceptual neglect and representational neglect, and the phenomenon of implicit processing of information in the neglected hemifield. In so doing we explore the implications of the findings from this literature for the development of theories of visuo-spatial working memory. PMID- 8713548 TI - Imagery and perception-action mediation in imitative actions. AB - This paper describes two lines of research exploring a hypothetical function of imagery in the context of imitative actions: the mediation between perceptual and motor processes. Both experimental approaches, a sequence learning task and a timing imitation task, demonstrate that engagement into imagery as a temporally distinct activity between observation and performance is not required for accurate imitation. Moreover, evidence is provided that generative processes can take place during event observation itself, thus making a separate recoding stage redundant. Nevertheless, in the absence of a visual display, imagery of a movement sequence exerted similar learning effects as physical and observational practice, and visual and motor imagery were found to be equally effective rehearsal strategies for maintenance of temporal information in short-term memory. PMID- 8713549 TI - Do imagined and executed actions share the same neural substrate? AB - This paper addresses the issue of the functional correlates of motor imagery, using mental chronometry, monitoring the autonomic responses and measuring cerebral blood flow in humans. The timing of mentally simulated actions closely mimic actual movement times. Autonomic responses during motor imagery parallel the autonomic responses to actual exercise. Cerebral blood flow increases are observed in the motor cortices involved in the programming of actual movement (i.e. premotor cortex, anterior cingulate, inferior parietal lobule and cerebellum). These three sources of data provide converging support for the hypothesis that imagined and executed actions share, to some extent, the same central structures. PMID- 8713550 TI - Ideomotor apraxia and cerebral dominance for motor control. AB - Ideomotor apraxia is a symptom of left hemisphere damage. Patients with ideomotor apraxia commit errors when imitating movements with their left, non-paralyzed hand. This has been taken as evidence for a motor dominance of the left hemisphere. It has been hypothesized that the left hemisphere is dominant for internal preprogramming of skilled movements of either hand. We investigated the kinematics of movement trajectories of imitation of meaningless gestures. Group analysis confirmed that hesitant, feedback-controlled movement prevail in patients with apraxia, but analysis of single cases revealed the existence of kinematically normal movements leading to apractic errors. Enhanced reliance on feedback-control appears to be a compensatory strategy rather than the source of apractic errors. In a second study we explored the alternative hypothesis that patients with apraxia lack a general concept of the human body which is necessary to mediate the translation of a target position seen on the model into a target position on the patient's body. Imitation of movements was examined on oneself and on a mannikin. Patients with apraxia who made errors when imitating on themselves committed errors also when imitating on the mannikin. Taken together, both studies support the view that the source of errors in the imitation of gestures is to be sought at a conceptual level. This casts doubts on the alleged dominance of the left hemisphere for motor control. PMID- 8713551 TI - The role of inhibition in the hierarchical gating of executed and imagined movements. AB - A theory is presented concerning the neuronal mechanisms which may underlie the organisation of imagined versus executed movements. A review is first presented of previous theoretical and experimental evidence suggesting that the brain can use the same mechanisms for the imagination and the execution of movement. In particular the fact that adaptation of the vestibulo-ocular reflex can be obtained by pure mental effort and not solely by conflicting visual and vestibular cues has been suggestive of the fact that the brain could internally simulate conflicts and use the same adaptive mechanisms used when actual sensory cues were in conflict. The saccadic system is taken as a good model for the study of this question because the mechanisms which underlie saccade generation are now partially understood at different levels from the brain stem to the cortex. The central idea of the theory is based upon the fact that, in parallel with the excitatory mechanisms underlying saccade generation, several inhibitory mechanisms in cascade allow the selective modulation and blockage of saccades. Synaptic inhibition is therefore supposed to play a major role in a hierarchical selective gating of saccade execution not at one but at several levels allowing a variety of different types of "imagined movements' some involving only the higher levels some in which the execution is only blocked at the very immediate premotor level. But in all cases the theory proposes that imagination and execution have many mechanisms in common. PET data showing that indeed the same structures are activated in both types of movements support this idea although the final answer will have to be brought by neuronal data. PMID- 8713552 TI - EEG aspects of mentally playing an instrument. AB - This pilot study examines the possibility to detect activities of SMA by means of EEG coherence analysis in a female professional violoncellist. The proband was asked, for 5 min each, to listen to a piece of music (she knew by heart), to imagine playing this piece and to imagine playing scales. The experiment was repeated after 5 days. Consistent significant coherence changes with respect to the averaged EEG at rest were plotted as probability maps. For each of these three tasks different coherence patterns emerged. Among the electrodes next to SMA, Fz was most involved while playing scales, less while imagining playing the same piece and still less while just listening to it. PMID- 8713553 TI - Electric and magnetic fields of the brain accompanying internal simulation of movement. AB - Methods of functional brain imaging have been used to identify brain structures which are active during internal simulation of movements (ISM). Between 1977 and 1993 it was consistently reported that the primary motor cortex (MI) is not active during ISM whereas other cortical areas, in particular the supplementary motor area (SMA) are active. ISM was assumed to be a situation of "internal programming'. Brain systems involved in ISM or 'programming' were hypothesized to be superior to and separable from 'executive system' including MI. We have studied electric and magnetic fields of the brain when subjects internally simulated either a single movement or a sequence of movements. Results of the studies are consistent with the assumption that MI is active with ISM. Internally subjects experienced effort which was required to inhibit overt movements during ISM. A recent EEG study showed different patterns of cortical activity with ISM and with movement inhibition suggesting that different brain structures may be active during ISM and movement inhibition [23]. PMID- 8713554 TI - Premotor cortex and the recognition of motor actions. AB - In area F5 of the monkey premotor cortex there are neurons that discharge both when the monkey performs an action and when he observes a similar action made by another monkey or by the experimenter. We report here some of the properties of these 'mirror' neurons and we propose that their activity 'represents' the observed action. We posit, then, that this motor representation is at the basis of the understanding of motor events. Finally, on the basis of some recent data showing that, in man, the observation of motor actions activate the posterior part of inferior frontal gyrus, we suggest that the development of the lateral verbal communication system in man derives from a more ancient communication system based on recognition of hand and face gestures. PMID- 8713555 TI - Comparison of neuronal activity in the supplementary motor area and primary motor cortex. AB - Neuronal activity in the supplementary motor area (SMA) and primary motor cortex (MI) have been compared in many experiments during performance of many different motor tasks. On one hand, the activity in both areas may appear similar, especially when the motor task is simple. On the other hand, if the motor tasks are more demanding, neuronal activity in the SMA exhibits a variety of complex relationship to many different aspects of motor behavior, while the activity in MI is mostly related to execution of motor task itself. Of particular interest is the neuronal activity in the SMA during preparation and execution of motor tasks when no external cues for the retrieval of appropriate motor act is available. Temporal sequencing of multiple movements is a typical example of the kind of motor task that requires profound activity in the SMA. PMID- 8713556 TI - On the translation of directional motor cortical commands to activation of muscles via spinal interneuronal systems. AB - I discuss in this paper some of the neural mechanisms by which directional motor cortical commands could be potentially translated into multi-muscle activations to generate a directed force (and initial movement) in space. Specifically, I review the results of recent studies in the motor cortex of monkeys and the spinal cord of the frog, and propose a possible mechanism by which these results could be formally connected. It is suggested that spinal mechanisms of the kind described in the spinal frog could serve as substrates for the operation of directionally tuned motor cortical activity to produce an appropriately directed motor output by the limb. PMID- 8713557 TI - Detrusor contractility in hyperreflexic bladders (from intracerebral neuropathies) versus unstable and stable unobstructed ones. AB - Three male patient groups of 20 subjects each have been assessed urodynamically by pressure flow study, namely, patients with detrusor hyperreflexia (DH) due to intracerebral neuropathies were compared with subjects bearing a condition of idiopathic detrusor instability, as well as with a control group of males showing normal urodynamic findings. Detrusor contractility proved greater in the unstable patients, but did not differ significantly between those with DH and the controls. Such results may suggest that intracerebral neuropathies leading to DH have no influence on, whilst the factors responsible for detrusor instability are strongly related to, the degree of activation of bladder muscle mechanical capability. PMID- 8713558 TI - Effect of intravenous alfuzosin on urethral pressure in patients with neurogenic bladder dysfunction. DORALI Study Group. AB - In order to assess the ability of a single intravenous (i.v.) injection of alfuzosin, a selective alpha-1 blocker, in reducing high urethral tone in patients with symptomatic neurogenic bladder dysfunction (NBD), 163 patients (mean maximal urethral pressure [MUP] 108 +/- 46 cm H2O) were enrolled in a double-blind, placebo-controlled, parallel-group trial and were randomly allocated to receive 0.5 mg (n = 45), 1 mg (n = 41), 2 mg (n = 39) alfuzosin or placebo (n = 38). The decrease in MUP was dose-dependent and statistically significant (P < or = 0.05) for 1 and 2 mg alfuzosin (respectively, 43 +/- 28 cm H2O and 46 +/- 27 cm H2O decreases vs. baseline) in comparison with placebo (23 +/- 30 cm H2O). The 2 mg dose level was the most effective leading to a > or = 30 or 50% decrease in MUP in, respectively, 69 and 44% of patients. The safety of all three alfuzosin dose levels was satisfactory and comparable to placebo. I.v. alfuzosin induces, in a dose-related manner, a clinically significant decrease in urethral pressure in patients with NBD and high urethral tone, and may be safely used as a pharmacological test as part of an urodynamic investigation. PMID- 8713559 TI - Maximal electrical stimulation in children with unstable bladder and nocturnal enuresis and/or daytime incontinence: a controlled study. AB - The aim of this study was to investigate clinical and urodynamic effects of anal MES in children with unstable bladder and micturition problems (nocturnal enuresis and/or daytime incontinence). Seventy-three girls, aged 5 to 17 years, mean age 9.7 years, with cystometrically proved idiopathic detrusor instability and nocturnal enuresis and/or daytime incontinence, were treated by maximal electrical stimulation (MES) for 1 to 2 months. Twenty-one girls, aged 6 to 14 years, mean age 9.3 years, with unstable bladder and micturition problems used only the anal plug without a battery for 1 month and served as the control group. Four and a half months (1-36 months) after the end of treatment, 75% of the stimulated patients were cured or improved by 50% or more. In the control group, 86% of the girls remained unchanged (P < 0.01). One month after the completion of anal MES the average number of monthly nocturnal enuretic episodes fell from 14 to 6.5 (P < 0.001) and the number of daytime incontinence episodes diminished from 3 to 0 (P < 0.001). On an average of 14.5 months after the end of anal MES, enuresis recurred in 20% of cases. Post-MES cystometry showed intensified first desire to void (P < 0.05), as well as an increase in maximum cystometric capacity (P < 0.0001), bladder compliance (P < 0.0001), and volume of the first detrusor contraction (P < 0.01). A statistically significant decline in the number of uninhibited contractions was also noticed (P < 0.001). In the control group, the anal plug did not produce any significant cystometrical changes. Anal MES can be recommended as an effective method for treating nocturnal enuresis and/or daytime incontinence and unstable bladder in children. PMID- 8713560 TI - Comparison between urinary diaries completed with and without intensive patient instructions. AB - The evaluation of the incontinent patient relies on accurate assessment of urinary symptoms. Although the 7 day urinary diary is a reproducible method of data collection, the optimal means of implementing this diary is unknown. The urinary diary is usually employed after the initial clinical pathophysiologic evaluation has been performed and the patient has received intensive instructions on the correct method of diary completion. This study aims to determine if a urinary diary provided to the patient prior to the initial clinical evaluation along with minimal instructions will provide symptom data comparable with that obtained by conventional methods. Two hundred seventy-eight women were recruited to participate in one of three clinical trials for urinary incontinence treatment. All subjects completed a diary prior to the initial clinical evaluation, the Minimal Instruction Diary, and a second diary after clinical evaluation, the Intensive Instruction Diary. The Minimal and the Intensive Instruction Diaries were compared for number of episodes of diurnal and nocturnal voluntary micturition and incontinence. Pearsons' correlation coefficients ranged from 0.67 to 0.78 for each of the urinary symptoms. Intra-subject comparison indicated a decline in reports of nocturnal voluntary micturitions from the Minimal to the Intensive Instruction Diary. No demographic or urodynamic parameters could account for the difference. The 7 day urinary diary is a reliable tool to asses urinary symptoms, which can be utilized prior to the initial clinical evaluation. Its case of use and practicality make this diary promising for use in a wider patient population. PMID- 8713561 TI - Laparoscopic implantation of temporary electrodes for bladder stimulation in dogs. AB - Five female mongrel dogs were used in an acute animal experiment. Under general anesthesia two monopolar wire electrodes armed with needles on either side were implanted into the bladder wall, the leads of both electrodes were then pulled through the abdominal wall. Filling of the bladder and intravesical pressure measurement were achieved through a suprapublic percutaneous cystostomy. Electrostimulation was carried out using the XEJ-2 experimental electroejaculator (biphasic pulses, pulse width 2.5 msec, approximately 70 mAmp, 50 Hz). The pressure changes within the bladder following electrostimulation were recorded, the effect of stimulation was observed endoscopically. Electrostimulation of the bladder wall resulted in micturition in all cases, showing an initial pressure peak at the commencement of electrostimulation followed by a decrease after onset of evacuation. We believe that laparoscopic implantation of temporary wire electrodes followed by percutaneous electrostimulation may open up new possibilities for bladder rehabilitation as well as for diagnostic investigations into the contractile capabilities of the bladder. PMID- 8713562 TI - Effects of long-term partial outflow obstruction on bladder function in the rat. AB - The effects of chronic partial outflow obstruction in rats were investigated. The urethra of male rats was partially obstructed for 3 or 6 months and bladder function was compared with that of age-matched controls. Bladder function was studied in vivo by infusion cystometry and in vitro by measuring the response of bladder muscle strips to stimulation. Cystometrograms of outflow-obstructed bladders were categorized into three types: type 1 was equivalent to a normal bladder; type 2 was characterized by large capacity, enhanced voiding pressure, and some residual urine; type 3 had the largest capacity, an impaired voiding pressure, and considerable residual volume (overflow-type of incontinence). The type 3 bladder was the most frequently observed type in rats obstructed for 6 months. Bladder weight increased significantly in rats with outflow obstruction. When five in vivo cystometric parameters (pressure at which micturition was induced, capacity, maximum voiding pressure, voided urine volume, and residual urine volume) were analyzed according to duration of obstruction, only two parameters (capacity and residual urine volume) in rats obstructed for 6 months differed significantly from those in age-matched controls. Evaluation of these values according to cystometric type showed a significant deterioration in four of five parameters in type 3 bladders. Contractile responses of the bladder in vitro to field stimulation, bethanechol, ATP, and KCI were significantly impaired in those obstructed for 3 or 6 months. When in vitro responses were analyzed according to the classification of cystometric type, deterioration of contractility was confirmed in both types 2 and 3 bladders. The present animal model of outflow obstruction can serve as a model of benign prostatic hyperplasia in humans. PMID- 8713563 TI - Cryptorchidism and infertility: retrospective study of 18 post puberal patients affected with monolateral cryptorchidism. AB - The Authors report their experience on patients with monolateral inguinal cryptorchidism, comparing the clinical picture with the morphological pattern of both the testicular parenchima and the semen. Light and electron microscopic analysis of testicular biopsy and spermatozoa from the ejaculate have been taken into account. 4 patients out of 18 (22.2%) showed a normal spermiogram, while 14 showed a pathological spermiogram. The morphological study of the testicular parenchima showed a spectrum of lesions, most of which advanced, confirming that they are irreversible lesions of the germinal epithelium. PMID- 8713564 TI - [Extrinsic ureteral endometriosis: diagnostic laparoscopic contribution]. AB - A case of extrinsic ureteral endometriosis diagnosed with laparoscopy is described. Endometriosis, a common disease in premenopausal women, should be considered in differential diagnosis in patients with ureteral obstruction. Usually, the diagnosis of ureteral involvement is made at surgery. Laparoscopy is, therefore, an important diagnostic aid in women with distal ureteral obstruction not attributable to calculous. PMID- 8713565 TI - [Leydig cell tumor of the testis]. AB - It represents a rare and mostly unilateral pathology with benign istological patterns. However, since the biological behaviour of the tumor is not foreseable from the istological characteristics, the follow-up must be prolonged. The treatment of this cancer is orchidofuniculectomy, sometimes associated, in selected cases, with retroperitoneal lymphadenectomy. In presence of tumor of small dimensions, above all if bilateral ones, it may be indicated a conservative treatment. Authors report six cases of Leydig cell tumors of the testis observed in sixteen years. All patients had orchidofuniculectomy by inguinal approach. In only one case has been associated retroperitoneal lymphadenectomy for the suspect of lymphatic metastasis. Follow-up of 3 months to 15 years is available for all patients: no one has died for the tumor and all are without signs of disease. PMID- 8713566 TI - [A possible therapeutic improvement of the antibacterial effect by monitoring the production of leukocyte free oxygen radicals during antibiotic treatment of infertile patients with chronic bacterial prostatitis]. AB - In the male chronic accessory gland bacterial infections (AGBI), antibiotic treatment (AT) efficacy usually evaluates the antimicrobial outcome through one or more spermiocolture (SC) become negative. Recently, bacterial olipeptide fMPL has been used to detect a specific Radical Oxygen Species production by leukocytes (L-RLO), even when they are present at low concentrations in sperm fractions specimens. In the male AGBI could be present endogenous fMLP at sufficient levels to produce L chemiotaxis in the semen, and secondly active their specific L-RLO production, till when bacteriospermia remained positive. In the Percoll 50% (Pc50%) fraction, at higher L concentrations, from 22 infertile patients affected by chronic bacterial prostatitis (BP) and enrolled to a randomly AT treatment with a Quinolone (n = 12) or a Macrolide (n = 10), through two secondary milestones (possible elevated basal L-RLO in the pre-treatment, T0; significant changes in the basal and fMLP-stimulated L-RLO production within each treatment group and between groups), we would verify if a normalized L-RLO production could be taken as break-point for the AT withdrawal before checking SC for control. Indeed, in T0 all patients had positive SC and exhibited both basal and fMLP-stimulated L-RLO levels higher than those observed in 2 control groups (group cA = 10 fertile men, without chronic BP; group cB = 10 patients affected by chronic abacterial prostatitis (AP). On the 3rd AT cycle for 14 days, together in 20/22 AT-treated patients, basal and fMLP-stimulated L-RLO levels become low or normal, as well as their SCs become negative (CFU/ml = 0), whilst in a third control group (group cC) of 10 not-AT-treated BP patients, through matched-follow up observations, these L-RLO values were always elevated and their SC remained positive (CFU/ml > or = 10(5)). In chronic BP patients, AT seems to demonstrate both antimicrobial effectiveness and reduction of L-RLO production with values similar to those of control group cA. The monitored L-RLO values during AT could be useful in order to ottimize antimicrobial effect: this tool being able to previse SC outcome, could be assumed to define clearly AT break-point and/or cycle numbers. PMID- 8713568 TI - Urethral sleeve advancement in repairement of distal hypospadias. AB - Thirtyfive boys with distal hypospadias were operated on using urethral sleeve advancement procedure between 1990 and 1994. Early complications such as hematoma and wound infection, and late complications such as fistula formation and meatal retraction were not seen in the patients except meatal stenosis in first month control in only eight patients. Four of them underwent dilatation and meatotomy was performed in the rest. Both cosmetic and functional results were excellent in the patients in sixth month control. PMID- 8713567 TI - Isthmus hypertrophy mimics tumour in horseshoe kidney: a case report. AB - We present a case of horseshoe kidney. He was operated because of nephrolithiasis, and was found to have a tumour-like mass in the isthmic localization 2 years after the operation. The tumour-like mass was proven to be an isthmus hypertrophy after detailed research. PMID- 8713569 TI - Renal oncocytoma. Pathological evaluation and clinical implications. AB - Renal oncocytoma is a benign tumour of renal tubular origin; oncocytes are transformed epithelial cells rich in mitochondria, probably representing senescent degenerative cellular changes. Most of renal oncocytomas usually follow a benign clinical course and partial nephrectomy or enucleation has been advocated as curative. By immunohistological staining of tissue sections using monoclonal antibodies (DBA, SBA, PNA, UEA, Cytocheratine), we can suppose the histogenetic origin of renal oncocytomas from a region other than the proximal tubular epithelium, and in particular from the collecting duct epithelium. We believe that it is most important to perform flow cytometry to study the chromosomal pattern of the tumour, once intra-operative frozen sections have advanced the suspicion of renal oncocytoma; if oncocytic cells show a diploid pattern, and the tumour mass is well circumscribed and has not an excessive diameter, we favour renal sparing surgery. PMID- 8713570 TI - Three consecutive cases of adenomatoid tumour of the epididymis: histological considerations and therapeutical implications. Review of the literature. AB - Adenomatoid tumour is an uncommon neoplasm of the male genital tract. The Authors report their recent observation of three consecutive cases of adenomatoi tumour of the epididymis. In all cases the diagnosis was difficult and done only by an accurate histological examination. The treatment was conservative, consisting in the removal of the neoplasm. After a follow up of at least two years the patients do not show any sign of relapses confirming the validity of the surgical therapeutical approach performed. Because of the unfrequent observation of these forms and the difficulty in the diagnosis, the Authors stress the necessity of the surgical exploration with an accurate microscopic examination of the specimens. Moreover a review of the literature from a histological and therapeutical point of view was done and reported. PMID- 8713571 TI - [Our experience concerning scrotal traumatology]. AB - The Authors present their experience on 24 blunt scrotal traumas observed since January 1991. They underline the importance of an ecographic scanning that enables a precise evaluation of the traumatic event. Patients that present positive ultrasonic findings are promptly operated thus permitting bleeding and infection control. In this way gonadic function is restored and hospital care reduced. The diagnostic approach is presented, cases reported and results discussed. After a review of the literature on the topic they stress the role of Eco-Color-Doppler examination in scrotal blunt trauma for the intrinsic characteristics of precision, rapidity and non invasivity. This permits a nosologic evaluation of all types of blunt trauma and selects, for surgery, only those patients with a well definied diagnosis. PMID- 8713572 TI - Cell cycle regulatory proteins are targets for induced differentiation of transformed cells: Molecular and clinical studies employing hybrid polar compounds. AB - Considerable progress has been made toward elucidating the pathway of induction of terminal differentiation of transformed cells by hybrid polar compounds such as hexamethylene bisacetamide (HMBA). HMBA alters factors controlling G1-to-S phase transition, leading to G1 arrest and inhibition of DNA synthesis. Among the inducer-mediated changes, suppression of cyclin-dependent kinase cdk4, which may be required for phosphorylation of the retinoblastoma protein pRB and perhaps p107, is critical in the pathway of terminal differentiation. HMBA induces an increase in the level of p21 which inhibits cyclin-dependent kinase activity and, in turn, may cause cells to arrest in G1. p107 complexes with transcription factor E2F, which may alter E2F-dependent gene transcription. the relationship of the inducer-mediated changes in cyclins, cdks, cyclin-cdk inhibitors and transcription factors to the expression of differentiation-specific genes has not yet been established. The hybrid polar compounds are potent inducers of differentiation of a wide variety of transformed cells. HMBA has been shown to induce differentiation of neoplastic cells in patients. A second generation of hybrid polar compounds have been synthesized which are up to 1000 fold more potent than HMBA on a molar basis as inducers of murine erythroleukemia (MEL) cells and other transformed cells in vitro. The potential of these compounds as clinically useful inducers of differentiation of cancer cells is under study. PMID- 8713573 TI - Pharmacokinetics of all-trans-retinoic acid in Japanese patients with acute promyelocytic leukemia. AB - All-trans-retinoic acid (ATRA) was administered at 45 mg/m2 to 4 Japanese patients with acute promyelocytic leukemia (APL). A pharmacokinetic study revealed that the mean peak plasma concentration was 208 ng/ml and was reached at 150 min after ingestion. The mean area under the concentration x time curve (AUC) was 498 ng.h/ml. Two patients showed a good hematological response to ATRA, and they had higher peak plasma concentrations of ATRA and larger AUCs. In one patient, dose escalation of ATRA (90 mg/m2) increased the plasma concentration markedly. In another patient, the plasma concentration decreased markedly in the fasting state. A larger pharmacokinetic study is necessary to examine the influence of food on the absorption of this agent and an optimum administration schedule of ATRA in Japanese APL cases. PMID- 8713574 TI - Attenuation of cardiotoxicity of daunomycin using a complex with heparin. AB - Daunomycin (DM) is one of the most important antitumor agents. However, the cardiotoxicity of DM limits it's clinical use. We have made an ionic complex with heparin to decrease the cardiotoxicity. Cardiotoxicities of DM and DM-heparin complex were compared in hamsters. On the electrocardiogram (ECG), two of the four hamsters given DM showed the serious abnormality, bidirectional ventricular premature contraction, while the hamsters given DM-heparin or saline had no abnormalities. On pathological examination, cardiac tissue in hamsters given DM showed deposition of basophilic materials, mild eosinophilic change of myofibrils and microvascuolization, whereas no change was observed in hamsters given DM heparin complex or saline. Acute toxic effects on survival rates and body weights were more profound in DM-infused mice than in DM-heparin-infused mice. DM and DM heparin complex showed similar anticancer activity both in vivo and in vitro. Thus, the present study suggests that the DM-heparin complex may attenuate the cardiotoxicity of DM without affecting it's antitumor effect in humans. PMID- 8713575 TI - Analysis of circulating hematopoietic progenitors in patients with chronic renal failure under hemodialysis. AB - Circulating hematopoietic progenitors were analyzed in patients with chronic renal failure (CRF) under hemodialysis (HD) by methylcellulose culture containing interleukin-3 (IL-3) to clarify the differences in hematopoiesis between patients with and without CRF-associated anemia and between good responder whose hematocrit (Ht) was preserved in more than 25% under erythropoietin (Epo) treatment and poor responders whose Ht remained less than 25% even under Epo treatment. The numbers of peripheral blood (PB) erythroid burst-forming units (BFU-E) and granulocyte-macrophage colony-forming units (CFU-GM) in HD patients without Epo treatment, whose Ht levels were greater than 30%, were similar to those in normal subjects. However, these numbers in HD patients who required Epo treatment were significantly lower than those in normal subjects. The number of PB BFU-E in HD patients who showed a poor response to Epo was significantly lower than that in HD patients who showed a good response to Epo. The number of PB BFU E was well correlated with the number of PB CFU-GM in all groups of HD patients. There also existed a definite correlation between these numbers and the Ht levels in HD patients without Epo treatment, but not those in HD patients with Epo treatment. The sensitivity of PB BFU-E to IL-3 was lower in HD patients who showed a poor response to Epo than in the other HD patients and normal subjects. These findings indicate that hematopoiesis in HD patients with CRF associated anemia is suppressed in both the erythroid and myeloid lineage at primitive stages, and that the lower sensitivity of PB BFU-E to IL-3 in HD patients with a poor response to Epo may be associated with this poor response. In addition, the level of the serum transferrin receptor (sTfR) in HD patients without severe anemia was higher than that in normal subjects and HD patients who required Epo treatment, indicating that erythropoiesis in HD patients who do not require Epo treatment is more active than that in normal subjects and other HD patients. PMID- 8713576 TI - Clinicopathological characteristics of primary Ki-1 anaplastic large cell lymphoma. AB - The clinical findings and prognosis in 15 patients with primary Ki-1 anaplastic large cell lymphoma (ALCL) were analyzed and compared with those of patients with T cell and B cell lymphoma and Hodgkin's disease. Clinical data revealed lymphadenopathy in 13 patients (87%) and skin lesions in eight (53%). Other organic involvements were hepatomegaly in two patients (13%), splenomegaly in five (33%), and bone marrow involvement in three (20%). The rate of skin involvement was significantly higher than that in B cell lymphoma and Hodgkin's disease. In laboratory findings the gamma-globulin concentration was significantly higher than that in T cell lymphoma, and the erythrocyte sedimentation rate (ESR) was significantly higher than that in B cell lymphoma. Complete remission was achieved in 11 patients (73%) and the five-year relapse free survival was 27%. The overall survival was 4.0-69.8 months (mean 30.6 months). The mean survival was compatible with that of T cell lymphoma and was significantly shorter than that in Hodgkin's disease. Ki-1 ALCL can be distinguished from other lymphomas clinically as well as pathologically. Because Ki-1 ALCL is chemosensitive and the prognosis is as poor as that of T cell lymphoma, aggressive chemotherapy should be employed for the treatment of this disease. PMID- 8713577 TI - The inhibition of human factor VIIa-tissue factor by antithrombin III-heparin is enhanced by factor X on a human bladder carcinoma cell line. AB - Previous studies have shown that antithrombin III-heparin effectively inhibited the factor VIIa-tissue factor complex. Herein, we show that the neutralization of factor VIIa in complex with the cell surface tissue factor by antithrombin III heparin was markedly enhanced by plasma levels of factor X. Active site-mutated factor X (S376A factor X) and factor Xa previously inactivated with dansyl-Glu Gly-Arg-chloromethyl ketone were as effective as plasma-derived factor X in this reaction, indicating that the active site serine residue of factor Xa was not involved in this mechanism. Furthermore, Gla-domainless factor X had no effect in this system, emphasizing the importance of the factor X Gladomain in this reaction. Antibody experiments revealed that this effect was not due to trace levels of a tissue factor pathway inhibitor contaminating either the factor X or antithrombin III preparations. The presence of heparin in this system was essential, as deletion of heparin resulted in a factor VIIa-tissue factor neutralization rate essentially identical to that observed for antithrombin III alone. Plasma levels of factor IX also accelerated the inhibition of factor VIIa tissue factor by antithrombin III-heparin, although its effect was not as pronounced as that of factor X. Other vitamin K-dependent plasma proteins including protein S, protein C and prothrombin failed to augment the inhibition of factor VIIa-tissue factor by antithrombin III-heparin. Factor X did not enhance the neutralization rate of factor VIIa-tissue factor by antithrombin III heparin when a carboxyl-terminal truncated tissue factor construct (TF1-219) was used, even in the presence of mixed phospholipids. Our collective finding suggest that antithrombin III and factor X bind to heparin at distinct sites on the heparin molecule resulting in a transient ternary complex of antithrombin III heparin-factor X that represents the anticoagulant species. Factor X conceivably guides complex to a phosphatidylserine-rich site on the cell surface in close proximity to the factor VIIa-tissue factor complex and facilitates rapid neutralization of factor VIIa. Our findings also suggest that the effect of heparin on the regulation of the extrinsic pathway of blood coagulation may be more profound than previously recognized. PMID- 8713578 TI - Chronic neutrophilic leukemia with dysplastic features mimicking myelodysplastic syndromes. AB - A 52-year-old male patient with chronic neutrophilic leukemia (CNL) with dysplastic features is described. He had a 2-year history of anemia followed by marked leukocytosis up to 57.5 x 10(9)/l with 88% segmented neutrophils. Bone marrow aspiration and biopsy showed hypercellular marrow with myeloid hyperplasia and 16% myeloblasts. There were also significant morphological abnormalities which included neutrophils with few granules, hypersegmented nucleus or Pelger Heut anomaly, and micromegakaryocytes. Cytogenetic analysis disclosed a deletion of the long arm of chromosome 7 (7q-). He was diagnosed as having CNL with dysplastic features and was treated conservatively. However, leukemic transformation to acute myelogenous leukemia occurred within a year and he died 16 months after diagnosis. Neutrophilia is a feature not of myelodysplastic syndromes (MDS) but rather of myeloproliferative disorders such as CNL. However, this patient was considered to have MDS with increased proliferation and differentiation of neutrophilic lineage, because of marked myelodysplasia and poor prognosis. PMID- 8713580 TI - [Unilateral prolapse of the laryngeal pouch due to the deformity of the thyroid cartilage]. AB - Our examination included 10 male patients in the age of 40-72. Microlaryngoscopic examination revealed smooth prominence of the larynx vestibule always on the left side concealing partly the vocal fold but with its mobility preserved. Our greatest attention was focused on CT since we expected possible explanation of the asymmetry of the larynx vestibule. That examination disclosed: left-side shift of the thyroid cartilage lamina to the larynx, elongation of the thyroid cartilage lamina and measurements of interlaminar angles revealed contraction of that angle to the value 60-78. PMID- 8713579 TI - Aggressive CD5-positive diffuse large B cell lymphoma showing c-myc rearrangements developed in a patient with autoimmune hemolytic anemia. AB - A case of CD5-positive diffuse large cell lymphoma in a patient with autoimmune hemolytic anemia (AIHA) is reported. The patient was diagnosed with AIHA in December 1988. Three and a half years later, the patient complained of fever and left sided flank pain. Abnormal lymphocytes appeared in the peripheral blood and were positive for HLA-DR, CD5, CD19, CD20, and surface immunoglobulin (mu, lambda). The pathological diagnosis of the cervical lymphnode was non-Hodgkin lymphoma; diffuse large cell type with a starry sky-like appearance. Although the 8q24 translocation was not detected by karyotypic analysis of the peripheral blood mononuclear cells (PBMNC), Southern blot analysis revealed that the c-myc rearrangements had occurred. This case showed two rearranged bands with Eco RI, Bam HI, or Bgl II digestion, and a germline band with Hin dIII digestion using a second exon fragment of the c-myc gene as a probe. Despite intensive chemotherapy, this patient died 6 months after being diagnosed with malignant lymphoma. We discuss the c-myc rearrangements in this aggressive CD5-positive diffuse large B cell lymphoma. PMID- 8713581 TI - [Intranasal microsurgery in treatment of cerebrospinal rhinorrhea]. AB - The authors describe two female patients with CSF-leakage. The first patient showed spontaneous CSF rhinorrhea from a leak in the posterior wall of the sphenoid sinus and the second one showed a traumatic fistula in "lamina cribrosa" area. The treatment of choice is closure of the CSF leakage by nasal surgeon (intranasal and with help of binocular microscope under hypotensive anaesthesia) using either intranasal-transseptal or intranasal-transethmoidal approach). This preserves olfaction and avoids the risk of a neurosurgical or paranasal methods. The diagnosis of CSF rhinorrhea can be established by beta-2 transferrin test, isotope scanning or fluorescence endoscopy. The high-resolutions CT is the best method for localization of the bone defect. The intranasal interposition of fascial or conchal grafts with use of fibrigen glue in cases of small defect (< 1 2 cm) is always successful. PMID- 8713582 TI - [The usage of light density heparin (fraxiparin) in the treatment of orbital phlegmonous cellulitis with orbital veins and thrombophlebitis of the cavernous sinus]. AB - A case of 72 years old man, diabetic, suffering from phlegmonous cellulitis with orbital veins and sinus cavernous thrombophlebitis as a nasal vestibule furuncle complication is presented. Apart from the application of antibiotics and surgical procedure, the patient was treated from beginning with thrombolytics specimen, fraxiparine. The methods of treatment of cavernous sinus thrombosis with thrombolytic specimens evoke in the literature controversial opinions. In the author's opinion the early application of fraxiparine in presented case could be regarded as responsible for stopping the process orbital veins and sinus cavernous thrombosis and even causing it to recess. The complete cure without complications (e.g. blindness) is attributed by the authors to the anticoagulation treatment. The authors also stress that fraxiparine may by used for treatment of cavernous sinus thrombophlebitis even when the surgical procedure is planned. PMID- 8713583 TI - [The use of osseointegration of hearing aids]. AB - The principles of osseointegration defined by Branemark are reviewed. The authors describe the otological applications of titanium implants: bone anchored hearing aids (BAHA). The good results obtained depend on the selection of the patients and the observance of a rigorous surgical technique. The present paper summarizes our results with 32 BAHA implanted in 27 patients. PMID- 8713584 TI - [Comments on the clinical picture of Warthin's tumors]. AB - The author present a clinical analysis of 23 cases of Warthin's tumor treated in the I Clinic of Otolaryngology Silesian Medical Academy in years 1982-1991. In 16 cases the tumor was localised in the superficial part of the parotid gland, in 3 cases a multifocus tumor was found. The Warthin's tumor in 3 patients was localised in the deep part of the parotid gland. In one patient the tumor was beyond the parotid gland. PMID- 8713585 TI - [The treatment of subglottic defect with the help of myoperisteal flap from sternum]. AB - Reconstruction of subglottic defect with the help of sternocleidomastoid myoperiosteal flap was performed in 6 patients. Four of them were successfully decannulated. The average time from reconstruction to decannulation was 67 days. Technic of operation was described and discussed in light of our own and literature experiences. PMID- 8713586 TI - [Juvenile angiofibroma as a diagnostic and therapeutic problem (from the materials of the ENT clinic in Warsaw in 1980-1990)]. AB - Author described results of surgical treatment of 28 patients with histologically proven juvenile angiofibroma treated at the ENT Clinic of Warsaw Medical Academy between 1980 and 1990. Histology, morphology and clinical course of this rare tumor was shortly discussed as well as current diagnostic and therapeutic methods. PMID- 8713587 TI - [Head and neck tuberculosis: a still urgent problem]. AB - The upper respiratory tract is an unusual site for tuberculous infection. Most of the cases are secondary to active pulmonary tuberculosis. We present thirty five cases of tuberculosis localize in the head and neck region. There were twelve patients with lymphonodular tuberculosis, eleven patients with laryngeal tuberculosis, six patients with oral and pharyngeal tuberculosis, three patients with partoid gland tuberculosis, two patients with nose and paranasal sinuses tuberculosis and one patient with middle ear tuberculosis. This cases exemplifies the difficulty in diagnosis of tuberculosis in such an unusual sites. PMID- 8713588 TI - [The point of view of otolaryngologist and head and neck surgeon on thyroid diseases]. AB - In the paper authors have analyzed 70 cases of different neck, throat and larynx tumors connected with thyroid gland (44 female and 26 male, age 14 to 78), treated surgically in ENT Department of Country Hospital No 1 in Rzesow from 1974 to 1994. The material consists of 35 (50%) patients with neck tumors of thyroid origin, 20 (28,6%) patients with the tumor of thyroid gland or with nodular goitre, 8 (11,4%) patients with advanced cancer of the thyroid with shortness of breath and 7 (10%) patients with ectopy or splitted of thyroid. Based on the histopathological examination it was revealed that the highest group consists of patients with thyroid cancer because all of them live in the south-east part of Poland, which is known to be the area of higher risk of thyroid cancer. Neck tumor was the first symptom of thyroid disease in half of patients from the group mentioned above and the first symptom of 57,5% of patients with malignant neoplasms of thyroid gland were metastases to the neck lymph nodes. Among patients with thyroid malignant neoplasm the male:female ratio was 1:1,35, and the mean age-53,5 years was about 10 years higher than in the rest of patients. In 7 patients with thyroid cancer effective radical surgical treatment was applied. In 5 cases surgical treatment exceeded thyroid gland. It required very careful dissection and identification of recurrent laryngeal nerves and parathyroid glands to avoid iatrogenic damage. PMID- 8713589 TI - [Side effects of drug on the central part of the vestibule]. PMID- 8713590 TI - [Current issues of pediatric audiology. Part I. Etiology and diagnosis of congenital and perinatally acquired hearing disorders]. AB - The results of present studies on etiology, time of diagnosis and fitting of hearing aids in Germany are described. Possibilities of hearing screening examination and basic clinical procedures in hearing loss diagnosis are presented. PMID- 8713591 TI - [A case of glomus caroticum malignum]. AB - In the paper by the case of glomus caroticum malignum of a woman aged 54 was presented. The tumor was taken out operatively without any difficulties by splitting mandibulae and cutting the muscles of the mouth bottom with wide exposure of the parapharyngeal space. This way of operations is considered and proved us a practicable method concerning the localization of the tumor. PMID- 8713592 TI - [The application of microspot micromanipulators in carbon dioxide laser surgery of laryngeal cancer]. AB - The authors present the advantages of using a micromanipulator of new generation (small working spot) in laser surgery of larynx. This micromanipulator are very usefull in laser surgery of small changes on vocal cords. PMID- 8713593 TI - [Xanthogranuloma juvenile of the maxillary sinus]. AB - The authors describe a rare case of xanthogranuloma juvenile of a nose and maxillary sinus in 3 years old girl. The tumor was situated in the left nasal cavity and the left sinus. Surgical treatment was done. Histological examination disclosed xanthogranuloma juvenile. The duration of follow up was 7 months without any sight of recurrence. This report documents the first known case of maxillary sinus involvement. PMID- 8713594 TI - [ENT department on the 50th anniversary of the Academy of Medicine in Gdansk]. PMID- 8713595 TI - [Nineteenth century physicians against drum perforation]. AB - The trials of "organic" closure of drum perforation are described. The achievements of Adam Politzer, Hermann Schwartze, Joseph Gruber are presented. The first who used term "myringoplasty" was Emil Berthold. The "epochal" method of Wasilij Okuniew and achievements of Beniamin Gomperz are also depicted. The scientific activities of Polish otologists: Ludwik Guranowski and Rafai Spira were presented. PMID- 8713596 TI - [Hand support for nasal endoscopy and functional endonasal surgery]. AB - The authors present hand support for nasal endoscopy and functional endonasal surgery which prevents fatique of the surgeons hand during operation. The advantages of this device are that it provides precession and shorten time of operation. PMID- 8713597 TI - [Cytodiagnostics of the cancer and precancerous states of the larynx]. PMID- 8713598 TI - [Report on the 9th British Academic conference in otolaryngology (BACO), Manchester, July 9-14, 1995]. PMID- 8713599 TI - [Report on the 5th European craniofacial congress, Copenhagen, June 15-17, 1995]. PMID- 8713600 TI - [Report on the 68th congress of Czech otolaryngologists, Plzen, June 28-30, 1995]. PMID- 8713601 TI - [Clinico-epidemiologic profile of Chagas disease patients attending an ambulatory referral center and proposal of a model of care for the Chagas patient under the perspective of a comprehensive health care]. AB - A case-control study to analyse clinical and epidemiological data of 842 patientes seen in an outpatient ward reference center for chagas disease in Belo Horizonte, Brazil, from 1985 to 1992. It was verified that these patients were in a average age of 37.78 years; lower literacy regardless of race, and, performed activities that demanded greater muscle effort (p < 0.05). The main mechanism for disease transmission proved to be the vectorial route. Disease-especific complaints prevailed in the above mentioned patients (p < 0.05). The most important abnormalities found in clinical examination were with cardiac sounds (OR: 2.96 95% CI: 1.29-4.09) and the presence of extrasystole (OR: 7.16 95% CI: 1.59-45.07). Imunofluorescence Test was more sensitive (94%) and specific (96%). It also had a PPV of 99% and e NPV of 83%. ELISA test carried out in 43 patientes did not yield false results. Indeterminate chronic phase (56%) or early stages of the disease prevailed. The findings of normal EKGs and abnormalities in 30% of the Holter tests, 41% of the exercise tests, 33% of the ECO tests and, 48% of normal chest x-ray with abnormal ECO, high lights the importance of caring out more detailed exams in specific situations. We hereby propose a care model for the Chagas disease patient, integrated to our State Health Plan stressing serologic tests, and training capable human resources committed to the development of integrated health care and the optimization of our referral and counter-referral system, thus issuing total care to the Chagas disease patient. PMID- 8713602 TI - [Hospital morbidity for Chagas disease in Brazil]. AB - The aim of this research was to describe the delivery and evolution of Chagas disease with in-patients in Brazil during the last decade. It also attempted to discuss the limits and possibilities of Internal Hospitalization Authorization (AIH) as a source of epidemiologist information. From 1984 to 1993, the number of in-patients diagnosed with "tripanossomiase" was about 1,700 yearly, declining in absolute numbers during the entire historic series. The majority concentration of in-patients occuring in this study was recorded in Sao Paulo, Goias, Minas Gerais and Distrito Federal. The public sector importance increased and the university hospitals improved their participation with in-patients with "tripanossomiase". Concerning hospitals expenses for in-patients, this study has observed that a hegemony in Sao Paulo was strengthened. The average cost was varied: in Sao Paulo and Distrito Federal, the average cost was much higher than that of Minas Gerais and Goias. Finally, this study concluded that the most suitable usefulness of the AIH forms would be for disease surveys that haven't been presented to many clinics, for which the SINTESE files would be sufficient. PMID- 8713603 TI - [Evaluation of a program for the control of schistosomiasis in municipalities located on the basin of the Sao Francisco River, Minas Gerais, Brazil]. AB - An evaluation of the control program on schistosomiasis (PCE/PCDEN) was performed in the region of Sao Francisco river in MInas Gerais. The study area comprises six municipalities, with 130,000 inhabitants and 916 localities situated in an area with 10,722 km2. The activities initiated in 1983-85 in four municipalities and in 1987 in the other two. The main measures of control were repeated treatment with oxamniquine and use of niclosamide. The prevalence of infection by Schistosoma mansoni in the first four municipalities, that was initially around 18 and 32%, dropped abruptly after the first intervention (1983/85) and remained in levels below the initial ones until the last assessment (1990-94); similar trends were observed for the proportion of infected snails. In these municipalities, the proportion of localities without infection or with prevalence below 5% increased in relation to those with higher prevalence. In the other two municipalities, with initial prevalence below 5%, there were no substantial changes in S. mansoni prevalence or proportion of infected snails; the cost benefit of the program in these municipalities need to be assessed and the priorities redirected to eradicate focal areas and prevent spread to non infected localities. The authors call attention to the difficulties in the long term of a control program based on repeated treatments. Information on factors associated with S. mansoni infection in each locality, or in groups of similar localities, would allow to develop additional measures to treatment that could last longer and be less dependent on the continuous use of chemotherapy. PMID- 8713604 TI - [The Queixadinha Project: morbidity and control of schistosomiasis in an endemic area in the northeast of Minas Gerais, Brazil]. AB - In an endemic area for schistosomiasis in the northeast of the state of Minas Gerais in Brazil 516 individuals have been submitted to clinical and laboratory examination, ultrasonography of the abdomen and dopplerecocardiography in order to define the morbidity of schistosomiasis before and after treatment. A high prevalence of schistosomiasis (66.3%) and of severe disease (9.5% with palpable spleens) were recorded. Ultrasonography classified liver periportal fibrosis as light (19.4%), moderate (27.6%) and intense (6.8%), and 46.0% presented no periportal fibrosis. Twenty one out of the 53 individuals (39.6%) with palpable spleens did not present liver fibrosis on ultrasound. Periportal lymph nodes were described in 33.8% of the population and anti-KLH antibodies were found in the serum of 40.7%. Urinary alterations compatible with the glomerulopathy of schistosomiasis were observed in 4.5% of the population, and 11.7% of the individuals examined by dopplerecocardiography had pulmonary hypertension. Twelve months after treatment for schistosomiasis the prevalence of the disease dropped from 66.3% to 25.0%. In Queixadinha, a profile of the morbidity of schistosomiasis has just been established. PMID- 8713605 TI - [Serodiagnosis of Chagas disease: new reagent for the indirect hemagglutination test (THAI(IAL))]. AB - A new reagent was designed to the indirect hemagglutination test (IHATIAL), utilizing goose red blood cells as inert matrix and standardized for the field diagnosis of American trypanosomiasis. The objective was to substitute the lyophilized or frozen reagent of IHAT produced routinely using human erythrocytes in the Adolfo Lutz Institute (Sao Paulo/Brazil). The standardized reagent presented a long stability in liquid suspension, and was evaluated in 137 serum samples from patient with and without Chagas' disease, by IHATIAL. The diagnostic performance of this test was similar to the IHAT utilizing human erythrocytes and to that of a commercial IHAT kit. The sensitivity was 1.00, specificity 0.98, predictive value of positive 0.96 and of negative 1.00. Different batches of reagent successively produced proved to be reproducible in a quality control method. The new reagent is more economic than the former reagent, it can be produced easily and may be applicable to the seroepidemiologic studies. PMID- 8713606 TI - [Immunodiagnosis of schistosomiasis mansoni with a low parasitic load]. AB - Presently, the schistosomiasis mansoni with low worm burden is frequent, thus immunologic assays of interest for the field diagnosis of Schistosoma mansoni light infections were evaluated here. Assays not assessed before (group I) and those requiring better validation (group II) for the screening of light infections were included in this study. In the group I, the immunofluorescence assays for the detection of IgM antibodies to worm antigens (IgM IFAw) and IgG antibodies to egg antigens (IgG IFAe) gave high levels of sensitivity, specificity, efficiency and predictive value of positive. However, the immunoenzymatic assays for the detection of IgM antibodies to worm antigens (IgM ELISAw) and to egg antigens (IgM ELISAe) had lower levels than the former assays. The assays from the group II designed mostly for the detection of IgG antibodies to same parasite antigens showed good diagnostic performance. The data obtained here contributed to evidenciate at least three category of immunoassays, and we concluded that those from the category I are suitable for seroepidemiologic purposes by keeping their diagnostic features unchanged even varying significantly the intensity of S. mansoni infection. PMID- 8713608 TI - [Direct agglutination tests in the serodiagnosis of visceral leishmaniasis in the state of Para]. AB - The direct agglutination test (DAT) was evaluated for serodiagnosis of visceral leishmaniasis (VL) in human and canids (dogs and foxes Cerdocyon thous). The results were compared with those of the immunofluorescent antibody assay (IFAT) and enzyme-linked immunosorbent assay (ELISA). The sera used were from: humans (303): confirmed VL (16), suspected VL (65), other conditions (102), negative controls (15) and individuals from an endemic area (105); dogs (82): from an endemic area (68), Salvaterra/Marajo/Para (21 of which were parasitologically positive), and negative controls (14), from Belem; foxes (9): caught on Marajo Island. Antigens for DAT were prepared from promastigots of L. (L.) donovani, L. (L.) chagasi. Antigens used in ELISA and IFAT were prepared from promastigotes (soluble antigen) and amastigotes respectively of L. (L.) chagasi. In humans, the specificity and sensitivity of DAT using L. (L.) donovani were high (98.4% and 100% respectively) and comparable to that of IFAT (97.5% and 100%). ELISA was less specific (84.8%) although similarly sensitive (100%). In dogs, DAT was more specific using L. (L.) donovani as antigen than using L. (L.) chagasi. However, both DAT and ELISA were less sensitive (both 71.4%) than IFAT (100%). This difference was reflected in the results from endemic dogs, 87% of which were positive by IFAT but only 54% by ELISA and 49% by DAT. Similarly, all 9 fox sera were positive by IFAT, 7 of 9 (78%) by ELISA but none by DAT. In conclusion, DAT using L. (L.) donovani antigen can provide a useful test for human VL; utilization on a large scale would be possible with a suitable reference laboratory to monitor antigen quality. However, DAT appears less useful for canine studies, as it was less sensitive than ELISA and especially IFAT in detecting canine infection. PMID- 8713607 TI - The FML (Fucose Mannose Ligand) of Leishmania donovani. a new tool in diagnosis, prognosis, transfusional control and vaccination against human kala-azar. AB - The Fucose-Mannose Ligand (FML) of Leishmania donovani is a complex glycoproteic fraction. Its potential use as a tool for diagnosis of human visceral leishmaniasis was tested with human sera from Natal, Rio Grande do Norte, Brazil. The FML-ELISA test, showed 100% sensitivity and 96% specificity, identifying patients with overt kala-azar (p < 0.001, when compared to normal sera), and subjects with subclinical infection. More than 20% apparently healthy subjects with positive reaction to FML developed overt kala-azar during the following 10 months. In the screening of human blood donnors, a prevalence of 5% of sororeactive subjects was detected, attaining 17% in a single day. The GP36 glycoprotein of FHL is specifically reconized by human kala-azar sera. The immunoprotective effect of FML on experimental L. donovani infection was tested in swiss albino mice. The protection scheemes included three weekly doses of FML, supplemented or not with saponin by the subcutaneous or intraperitoneal routes and challenge with 2 x 10(7) amastigotes of Leishmania donovani. An enhancement of 80.0% in antibody response (p < 0.001) and reduction of 85.5% parasite liver burden (p < 0.001) was detected in animals immunized with FML saponin, unrespectively of the immunization route. PMID- 8713609 TI - [Evaluation of direct agglutination tests in the detection of Leishmania (Viannia) braziliensis in possible reservoirs of cutaneous American leishmaniasis in the state of Ceara]. AB - In order to evaluate the aplicability of the Direct Agglutination Test (DAT) as a method of detection of the natural canine infection for Leishmania (Viannia) braziliensis, it was realized a study involving the serum of naive dogs from Serra de Baturite, Ceara State, an endemic area of cutaneous leishmaniasis, and others from Curitiba, Parana State, a non endemic area. The results indicate the possibility of using this sorologic test to the epidemiologic research of the infection in the reservoir host; in this case, the dog. PMID- 8713610 TI - [Control of schistosomiasis infection in the localities of Cachoeira-Bahia, basin of the Paraguacu, 1982-1992]. AB - This study evaluates the effectiveness of the Schistosomiasis Control Program, PCE, in Cachoeira, state of Bahia-Brazil, in Paraguacu Basin Area. Data comes from PCE-111 cards of SUCAM/FNS, which are routinely registered in the 111 villages identified during the study follow up, 1982-1992. The S. mansoni infection was diagnosed through Katz-Kato technique for stool exams. Temporal trends of the prevalence of infection were analyzed for each village. Four trend patterns were identified, and the most common was those of recurrence of infection prevalence (67.6%). Control of the endemics was achieved in 22.5% of the villages, while expansion trends were found in only 9.9% of them. In general, there was found a decrease in the prevalence, particularly, from 1982 to 1985, followed by an increase that do not reach the initial levels. It was observed a relative reduction on the number of villages that shows prevalence below or equal to 5%, that indicates control of the infection. It was not possible to evaluate trends of infection intensity neither the occurrence of severe shistossomiasis disease or related deaths, that may allow a more complete evaluation of this program. PMID- 8713611 TI - [Morbidity of Chagas disease in areas of Sertao da Paraiba and Caatinga do Piaui]. AB - A clinical and electrocardiographic case control study was carried out with 186 pairs of persons with positive and negative serology for T. cruzi infection from the Sertao Paraiba and in 200 seropositive cases from the region of Caatinga in the State of Piaui, North-eastern Brazil. The predominant clinical manifestations in seropositive cases in both areas were: palpitations, dyspnea on effort, precordial pain, dysphagia, odynophagia, pyrosis and intestinal constipation. The EKG abnormalities rates suggestive of chronic chagasic cardiopathy were respectively in Paraiba and Piaui: AV block 3.8% and 2%, RBBB III 6.4% and 7%, RBBB III+ LAB 10.7% and 10.5%, and multifocal extrasystoles 2.7% and 3%. Xenodiagnosis in a sample of 54 seropositive individuals in the Sert-ao of Para iba and in 120 in the Caatinga of Piaui was revealed 13% and 34% positive; PCR tests in a sample of 47 seropositives in Paraiba and 101 in Piaui revealed positives in 44.6% and 59.5% respectively. Blood culture in LIT media of 101 seropositive cases from the Caatinga of Piaui was positive for T. cruzi in 25.7%. A triatomine survey carried out in a sample of 132 domiciles and peridomiciles in the Sertao of Paraiba and in a sample of 159 in the Caatinga of Piaui showed the following results: In Paraiba, 16 specimens of T. brasiliensis, not infected with T. cruzi, were captured. In Piaui, 750 triatomines were captured, of these 625 were examined: 49 were T. pseudomaculata, not infected with T. cruzi (19 in peridomiciles and 30 in the domiciles), and 576 were T. brasiliensis (371 in the domiciles and 205 in the peridomiciles) and of this latter specie 32 (5.5%) were infected with T. cruzi (31 in the domiciles and one in the peridomicile). PMID- 8713612 TI - [Preliminary results of a project about the ecology of Phlebotomus vectors of cutaneous leishmaniasis in the state of Bahia]. AB - Preliminary data of a project about the ecology of the Phlebotominae (sandflies) in a tegumentary leishmaniasis area of coconut plantation in the south of the State of Bahia, Brazil are presented. There are 60 dwellings far of one another, where existed 31 dogs and 229 inhabitants. Among them 41.5% were Montenegro positive; 37.5% from these had scars of healed ulcers and 8.8% had active ulcers. 53% of the house had dogs from which 22% were seropositive; from the 7 dogs with ulcers only 3 were seropositive. 14% of the inspected house harboured sandflies inside them. During two years of observations 72 sentinels hamsters were maintened at houses of patients with leishmaniasis ulcers, but they did not get the infection. During two years of observation, monthly collectings of sandflies were made inside house, chicken pen, curral, tree trunks and open field around a house with a patient with leishmaniasis. The following results were already obtained about the vectors: 5,614 specimens were identified as belonging to fourteen different species of sandflies. Among them Lutzomyia whitmani (92%) and Lutzomyia intermedia (4.8%) were the most abundant species. They are very anthropophilic and can be found inside houses and at peridomestic sites. Probably they are the principal vectors of the disease at domestic places. The other twelve species were less frequent and more found at sylvatic places in inspite they also bite the man. Generally the biting activity of most of the species of the area begins at 5 p.m. in the dusk and reaches its peak at 0 hour a.m., when begins declining until disapear at 7 a.m. L. whitmani was similarly collected with the same density in all lunar phases while L. intermedia was more abundant during the new moon phase. Most of the hundreds sandflies collected during the second year of observations, remains preserved in liquid nitrogen, watching for the adjustment of PCR molecular techniques to be processed for determination of the vector natural infection rates with leishmanias. Final results on all the project will be published as soon as the examination of such material has been processed. PMID- 8713613 TI - [Aminosidine in the treatment of mucocutaneous leishmaniasis caused by Leishmania (Viannia) braziliensis]. PMID- 8713614 TI - [Submental introduction of tracheal tube in endotracheal anesthesia. (Modified tracheal intubation method for cases of simultaneous maxillofacial injuries and osteotomies)]. AB - This article reports the modification of laterosubmental tracheal intubation in single-step surgery of panfacial fractures or osteotomies. In the modified method -after the submental transversal midline skin incision--the tube is guided through a submental tunnel to the oral floor, just behind to caruncula sublingualis. PMID- 8713615 TI - [Incidence of structural disorders of teeth in hereditary epidermolysis bullosa]. AB - Authors present a 12-year old boy, whose diagnosis was epidermolysis bullosa. Beside the general characteristics of the illness they discuss the dental status of the patient and sum up the literary references together with other anomalies occurring in epidermolysis bullosa hereditaria. PMID- 8713616 TI - [In vitro comparative study of instruments for root canal preparation. IV. Correlations between the original shape of the root canal and the asymmetry resulting from the preparation]. AB - Certain asymmetry of root canal preparation can be observed in the straight root canal group, as well. The modification of original root canal shape in this group usually is not substantial. Using hand instruments for shaping curved canals special tecniques are required to minimize the asymmetry. Though tendencies of different shaping characteristics can be observed on the mesiodistal and oro vestibular views there are no statistically significant differences between the asymmetries measured on the oro-vestibular and mesio-distal views. The three dimensional form of canals usually exclude the possibility of forming the same asymmetry on both projections, but the difference between the two views is not striking, therefore the conventional clinical radiographs usually do not give complete information about the asymmetry of preparation. PMID- 8713617 TI - [Social and behavioral factors influencing caries intensity and oral hygiene in an adult population in Budapest (sociological implications of a dental sociological survey)]. AB - Authors examined 868, 15-69 age old, non-selected patients. The findings of dental aspects of the investigation were reported in this periodical (88, 95-112, 1995). The sociological analysis verified strong connection between oral status and social stratification. The authors found that differences in oral status on the basis of educational qualification are larger than the differences on the basis of occupations. The effects of social factors on oral health behaviour are stronger than on oral status. The authors found strong connection between patients' oral health behaviour and parents motivations for patients' body culture. PMID- 8713618 TI - [Genetics of sexual development and its disorders]. AB - Different hypotheses for genetic control of sexual differentiation in humans are discussed. Information on structure and manifestation of genes determining development of internal and external male genitals is considered. The process of discovering the gene responsible for differentiation of the indifferent fetal gonad into testes is described. The etiological problems of sexual development disorders are considered on the basis of knowledge obtained by modern molecular and genetic methods. PMID- 8713619 TI - [Molecular cloning of mutant loci obtained by microinjection of oncogenic viral DNA into polar plasma of early embryos of Drosophila melanogaster]. AB - Mutations WRSV-1 and WRSV-4 obtained by microinjection of RSV cDNA into early embryos of Drosophila melanogaster were shown to be caused by the insertion of mobile elements jockey and B104(roo), respectively partial reversion of WRSV-lr arose due to the deletion of a 5'-region of mobile element jockey and of the adjacent region of the genomic DNA. It is concluded that genetic instability induced by oncoviral DNAs is based on the ability of these DNAs to induce transpositions of various mobile elements in the recipient genome. PMID- 8713620 TI - [Construction and analysis of transgenic plants of Nicotiana tabacum L. expressing a bacterial gene for beta-1,3-glucanase. I. Construction of vector plasmids for transfer into plants and expression of a modified gene for beta-1,3 glucanase from Clostridium thermocellum in tobacco protoplasts]. AB - We constructed two vectors, pC27-glc and pC29-glc, that allow expression of the beta-1,3-glucanase gene (glc) in plant cells. The glc gene was previously cloned from anaerobic thermophilous bacterium Clostridium thermocellum. To increase the efficiency of expression, the N-terminal fragment of the glc gene encoding bacterial transient peptide was deleted, and hybrid variants of lacZ-glc were obtained. Analysis of expression of the hybrid genes in Escherichia coli showed that deletion of the fragment corresponding to 31 amino acids (a.a.) of beta glucanase affected neither activity nor thermostability of the enzyme. The modified gene was subcloned into two vectors, pC27 and pC29, in which its expression was controlled by the TR2' promoter of the 2' gene of T-DNA and the rbcS promoter from Arabidopsis, respectively. Each of the resulting plasmids, pC27-glc and pC29-glc, was transfected into protoplasts of Nicotiana plumbaginifolia. Both the plasmids were shown to allow a high level of activity of the thermostable beta-1,3-glucanase. We plan to use the vectors obtained for transformation of agrobacteria and construction of transgenic plants. PMID- 8713621 TI - [Construction and analysis of transgenic plants of Nicotiana tabacum L. expressing a bacterial gene for beta-1,3-glucanase. II. Transgenic tobacco plants expressing the bacterial beta-glucanase gene from Clostridium thermocellum,--a model for studying the differential expression of stress response-related genes]. AB - The modified hybrid beta-1,3-glucanase gene (glc) of Clostridium thermocellum was expressed in tobacco Nicotiana tabacum. The glc gene was cloned into two plasmids, pC27-glc and pC29-glc, in which its expression was controlled by the TR2' promoter of the 2' gene of T-DNA and the rbcS promoter of Arabidopsis, respectively. These constructions were used for transformation of agrobacteria followed by transfer into plants. In transformed plants, each plasmid caused a high level of activity of thermostable bacterial glucanase not observed in reference plants. The plants obtained were used to study activation of some defense-related genes induced by their interaction with either tobacco mosaic virus (TMV) or a pathogenic fungus. PMID- 8713622 TI - [Genetic control of juvenile hormone metabolism in Drosophila melanogaster]. AB - Juvenile hormone (JH) metabolism in two lines of Drosophila melanogaster adults was studied under normal conditions and after short-time heat stress. Females of the line Cy/L; D/Sb were shown to have high JH hydrolysis levels which to decreased when they experienced heat stress by decreasing its level, Genetic analysis of differences in the level of JH metabolism in lines 921283 and Cy/L; D/Sb of D, melanogaster under normal conditions was performed. It was established that the trait studied was inherited according to the incomplete dominance type. It was shown that these differences were controlled by the gene (genes) localized in chromosome 2. PMID- 8713623 TI - [Genotoxic and mutagenic activity of antineoplastic anthracyclines and their aglycones: study in two test-systems]. AB - Mutagenic (Ames tests) and genotoxic (SOS chromotest) activities of highly efficient natural anthracycline monosaccharides possessing antitumor activity daunorubicin (also known as daunomycin or rubomycin), doxorubicin (adriamycin), and carminomycin-were studied. At the same time, the hypothesis was tested that intercalation of the antibiotic moiety into the helix of cell DNA, which was mediated by the saccharide amino group, played a crucial role in genotoxicity of these anthracyclines. The hydrolysis products of these antibiotics (the corresponding aglycones) and aclacynomycin A (an anthracycline trisaccharide), as well as aclavinone (its derivative aglycone), were studied. All these compounds lacked the saccharide amino group necessary for intercalation. It was found that all anthracycline monosaccharides studied had a strong mutagenic effect on strain TA98 and a moderate effect on strain TA100 of Salmonella typhimurium. Aclacynomycin A was found to have no mutagenic effect on any strain. Lack of the glycoside amino group did not necessarily result in loss of mutagenic activity in the derivative aglycones of anthracycline monosaccharides: they exhibited moderate mutagenic activity in strain TA98 and low but significant activity in strain TA100. The S9 microsomal fraction did not alter the mutagenic activity of either anthracycline monosaccharides or their aglycones; however, it dramatically increased the mutagenic activity of aclavinone: correspondence between positive responses in Ames tests and the SOS chromotest was found. Apparently, the mutagenic activity of the substances studied in bacterial cells was mediated by inducing the SOS-repair process. If the compound contained the amino glycoside moiety, functional and structural precursors of the SOS response were formed via intercalation of the reagents into the DNA duplex; if the substance did not contain this moiety, the precursors were formed via ionic interaction. PMID- 8713624 TI - [Beta-heterochromatin in Drosophila: molecular organization and function. Molecular biological analysis of a MAR/SAR DNA sequence in centromere heterochromatin of Drosophila melanogaster]. AB - A molecular biological characteristic of a DNA fragment delta 20p1.4, which is a moderately repetitive sequence of the Drosophila melanogaster genome, is presented. The fragment is present in about 120 copies per haploid genome. The main pool of the delta 20p1.4 homologous DNA can be isolated, along with the nuclear matrix DNA, and consists of Hind III-EcoR I monomers 1.4-1.6 kb in length. The monomers may occur in the genome as both single copies and tandem clusters forming chromosomes fragments up to 6-10 kb in length. The region of the delta 20p1.4 fragment between nucleotides 350 and 905 polymerizes with purified lamin from D. melanogaster. The sites of the fragment that had physical contact with lamin in vitro were determined using an Exo III protection. It was demonstrated that ATATTT, A, and T boxes located in four nonperfect tandem repeats were involved in the contact, both DNA strains reacting with lamin. PMID- 8713625 TI - [Genes of human viruses are less redundant than human genes]. AB - Statistical parameters of nucleotide sequences of mature human RNAs and those of human viruses were compared. The redundancy values of the appropriate genes were compared. The redundancy of virus genes was shown to be, on the average, less than that of human genes. The distribution of human genes according to redundancy values is bimodal, and that of human virus genes is trimodal. This fact suggests possibility of a novel gene classification according to statistical characteristics of nucleotide sequences. PMID- 8713626 TI - [Absence of DNA polymorphisms on the region of the APOV gene coding the proposed low density lipoprotein-binding domain of the ApoB-100 protein]. AB - To reveal the molecular basis of hereditary diathesis to hypercholesterolemia, single-strand conformation polymorphism (SSCP) analysis and subsequent sequencing were used to study structural organization of the APOB gene for apolipoprotein B 100 (ApoB-100). The gene region encoding a putative low density lipoprotein (LDL) receptor binding domain was analyzed in patients with myocardial infarction (MI). Differences in structure of the gene between patients with MI and healthy people were not detected. PMID- 8713627 TI - [Genetic sequelae of the accident at the Chernobyl nuclear power plant in house mice (Mus musculus)]. AB - Genetic disorders were studied in house mice caught from 1986 to 1993 in areas contaminated by radionuclides after the Chernobyl disaster. Dose rates on soil surface ranged from 0.02 to 200 mR/h. Frequency of reciprocal translocations in spermatocytes of the mice studied was relatively low, but increased with dose rate. In populations, frequency of mice heterozygous for recessive lethal mutations decreased with time after the accident. The data obtained allow us to assume that induced mutations may lead to elimination of germ cells and decreased viability in mice heterozygous for the mutations. These processes result in removing excess mutations from the population. PMID- 8713628 TI - [Genetic differentiation of helminths on the basis of data of polymerase chain reaction using random primers]. AB - The genetic variability of helminths belonging to the three classes Cestoda, Nematoda and Trematoda was analysed by means of polymerase chain reaction with randomly amplified polymorphic DNA (RAPD-PCR). Dendrograms of genetic relationships between some representatives of the genus Trichinella, Fasciola hepatica, Cysticerus ovis, Echinococcus granulosus, and Nematodirus spathiger were constructed on the basis of DNA amplification patterns. The formation of ecological and genetic adaptations in the helminth groups studied is discussed. PMID- 8713629 TI - [Effect of mutations on the dynamics of numbers of individuals with different genotypes in an isolated heterogeneous population]. AB - The overall (throughout the range) dynamics of the number of diploid individuals possessing n possible gene alleles in a given autosomal locus is studied in an isolated heterogeneous population. The significant influence of new mutations on overall dynamics is shown. It is demonstrated that, under certain conditions, the selection mechanism might have a greater effect on the dynamics of number of individuals with different genotypes in the isolated heterogeneous population than on mutation and "death-reproduction" mechanisms. PMID- 8713630 TI - Reflex sympathetic dystrophy revisited. PMID- 8713631 TI - Cancer pain. AB - An accurate assessment and where possible a precise diagnosis of the underlying mechanism in necessary for the effective management of pain in cancer. The majority of patients respond to oral analgesics, and the range of drugs required is not vast. Adjuvant analgesic drugs will be needed for the treatment of bone pain, visceral pain and neuropathic pain. For the remaining patients who present with difficult and occasionally intractable pain, a multidisciplinary approach using drug and non-drug measures will usually allow significant relief of symptoms. PMID- 8713632 TI - C.P.R. in the curriculum. PMID- 8713633 TI - Impact of anesthetic management on the outcome of cardiac patients for non cardiac surgery. PMID- 8713634 TI - Modified interface between pediatric ventilator and Ayre's T-piece for use in neonates. AB - The lowest tidal volume delivered by most of the pediatric anesthesia ventilators could be large for neonates, and may generate unacceptably high peak airway pressures. The effective tidal volume can however be reduced by connection of a small rubber bag between the ventilator and the expiratory limb of the Ayre's T piece circuit to divide the delivered gas mixture between the added bag and the neonate's lungs. The delivered tidal volume by the ventilator can then be manipulated to achieve the desired peak inspiratory pressure; 18-20 cm H2O in neonates with normal lung compliance. The modification was found to be simple, safe and convenient. PMID- 8713636 TI - Total intravenous anesthesia for diagnostic endoscopy in myasthenia gravis child- a case report. PMID- 8713635 TI - Diazepam serum concentration-sedative effect relationship in patients with liver disease. AB - Sedative effects and drug elimination characteristics after intravenous bolus administration of diazepam 0.15 mg/kg were investigated in 15 patients with liver disease (Group I), and in 15 normal patients (Group II) during diagnostic laparoscopy. Diazepam induced heavier sedation in patients with liver disease (P < 0.02). Serum concentration of diazepam 30 minutes after administration was significantly lower in Group I (group with liver disease) when compared with Group II (control group) (210.68 +/- 112.65 ng/ml vs 451.57 +/- 239.87 ng/ml, p < 0.02). The sedation scores during the laparoscopy procedures correlate negatively with serum benzodiazepine levels. The benzodiazepine concentration-time profiles of the groups vary significantly (p < 0.02). Heavier sedative effect and lower benzodiazepine concentrations in patients with liver diseases suggests that the permeability of blood brain barrier increases and that higher affinity to benzodiazepine receptors exists. Differences in concentration-time courses of diazepam in patients with liver diseases is a warning indicating the accumulation of the drug when infused or used frequently. PMID- 8713637 TI - Depression of excitatory effects of propofol induction by fentanyl. AB - Propofol is an intravenous anesthetic drug commonly used in outpatient anesthesia for its rapid and smooth onset of action, short recovery period and its minimal perioperative side effects. However, propofol like other intravenous anesthetic drugs can produce excitatory signs during induction without signs of epilepsy on EEG. We studied 64 patients scheduled for various outpatient procedures. The patients were allocated to 2 groups: Group A (n = 32) received on induction xylocaine 1 mg.Kg-1 followed by propofol 2 mg.Kg-1 over 20 seconds, Group B (n = 32) received xylocaine 1 mg.Kg-1 and fentanyl 1.5 ug.Kg-1 followed by propofol 2 mg.Kg-1 over 20 seconds. The incidence of excitatory movements (cough, hiccup, hypertonus, twitching or tremors) was in group A (13) significantly higher than that in group B (1), there was no significant difference between the two groups in the time of stay in the post anesthesia care unit. Fentanyl 1.5 ug.Kg-1 used in outpatient anesthesia with propofol 2 mg.Kg-1 provides cardio-vascular stability, deepens the anesthesia level, decrease the awareness and decrease the excitatory effects or propofol. PMID- 8713638 TI - Oral midazolam in pediatric dentistry. AB - The study was aimed at observing the changes in cooperation level of children undergoing dental procedures following the administration of a single oral dose of 7.5mg of Midazolam. Children were multinational, and ranged in age between 24 to 59 months. All subjects were considered to be resistant to routine behavioural management techniques. There was a 77.8% treatment visit success. The success rate was statistically insignificant between the sexes. Due to the small population, differences could not be elicited based on nationality and age group of subjects. There were however noticeable differences in behaviour based on the dental procedure being performed. Dental extraction had the poorest cooperation followed by cavity preparation. Cooperation improved significantly when the rubber dam was removed (where applied). Midazolam was found to increase treatment visit success rates in the present study. PMID- 8713639 TI - Effects of growth hormone and insulin-like growth factor I on salinity tolerance and gill Na+, K+-ATPase in Atlantic salmon (Salmo salar): interaction with cortisol. AB - The potential roles of growth hormone (GH) and insulin-like growth factor I (IGF I) in seawater (SW) acclimation of juvenile Atlantic salmon (Salmo salar) were examined. Compared to controls, fish in 12 ppt seawater given one or three injections (2-6 days) of GH (ovine, 0.2 microgram.g-1) or IGF-I (recombinant bovine, 0.05-0.2 microgram.g-1) had significantly greater salinity tolerance as judged by lower plasma sodium, osmolality, and muscle moisture content following transfer to 34 ppt. Single injections of GH and IGF-I in fish in fresh water failed to improve salinity tolerance following transfer to 25 ppt SW. Treatment of fish in 12 ppt with GH or IGF-I for 2-6 days did not increase gill Na+, K(+) ATPase activity, but treatment with GH prevented decreases in gill Na+, K(+) ATPase activity that occurred in controls following transfer to 34 ppt seawater. Fish in fresh water administered GH by implants (5.0 microgram.g-1) or osmotic minipumps (0.5 micrograms.g-1 day-1) for 7-14 days had greater gill Na+, K(+) ATPase activity and salinity tolerance than controls. IGF-I administered by implants (0.5-1.0 microgram.g-1) or osmotic minipumps (0.1 microgram.g-1 day-1) for 4-14 days did not increase salinity tolerance or gill Na+, K(+)-ATPase activity. Cortisol implants (50 micrograms.g-1) also increased gill Na+, K(+) ATPase activity and salinity tolerance after 14 days, and in combination with GH had a synergistic effect, Although IGF-I and cortisol implants had no significant effect after 7 days, in combination they significantly increased gill Na+, K(+) ATPase activity. The results indicate that GH and cortisol can increase salinity tolerance and gill Na+, K(+)-ATPase activity of Atlantic salmon and together act in synergy. Although IGF-I can increase salinity tolerance in short-term treatments (2-6 days) in 12 ppt, it is less effective than GH in increasing salinity tolerance and gill Na+, K(+)-ATPase activity in long-term treatments (7 14 days) and in interacting with cortisol. PMID- 8713640 TI - Isolation and structural characterization of proglucagon-derived peptides, pancreatic polypeptide, and somatostatin from the urodele Amphiuma tridactylum. AB - The expression of the preproglucagon gene in vertebrates is markedly species- and tissue-dependent. Three peptides derived from the posttranslational processing of preproglucagon were isolated from an extract of the pancreas of the urodele Amphiuma tridactylum (threetoed amphiuma). The primary structures of the peptides indicated identity with glucagon (HSQGTFTSDY10 SKYLDNRRAQ20 DFIQWLMST), glucagon like peptide-1 (GLP-1) (HADGTLTSDI10 SSFLEKQATK20 EFIAWLVSGR30 GRRQ), and glucagon-like peptide-2 (GLP-2) (HADGSFTSDI10 NKVLDTIAAK20 EFLNWLISTK30 VTE). Thus, in a urodele, as in the bullfrog but in contrast to the chicken and all nontetrapod species yet studied, pancreatic preproglucagon mRNA encodes a GLP-2 sequence. The amino acid sequence of glucagon has been better conserved during evolution of tetrapods (3 substitutions between amphiuma and human) than the sequences of either GLP-1 (7 substitutions) or GLP-2 (15 substitutions). Pancreatic polypeptide was also isolated from the extract and its primary structure (APKEPEHPGD10 DASPEQLEKY20 YQDLFQYIIF30 ITRPRY.NH2) indicates that the amino acid sequence of this peptide has been very poorly conserved, even among the amphibia. Amphiuma pancreatic somatostatin is identical to mammalian somatostatin-14. PMID- 8713641 TI - Elevations of somatolactin in plasma and pituitaries and increased alpha-MSH cell activity in red drum exposed to black background and decreased illumination. AB - The effects of different backgrounds and illumination levels on somatolactin (SL) concentrations in juvenile red drum (Sciaenops ocellatus) plasma and pituitaries were investigated using a homologous radioimmunoassay. Both plasma and pituitary SL concentrations were significantly higher in red drum exposed for 1 week to black backgrounds than in fish exposed to light backgrounds. Plasma SL levels decreased from 5.39 +/- 1.18 ng/ml in red drum adapted to a dark-blue background to 0.73 +/- 0.05 ng/ml within 1 hr of transfer to light-background tanks. Plasma SL concentrations were significantly elevated in fish 3 hr after transfer from light-background tanks (0.41-0.99 ng/ml) to black-background tanks (5.38 +/- 1.60 ng/ml) or black-background tanks without illumination (3.54 +/- 1.34 ng/ml). Maximum circulating SL levels (9.47 +/- 0.78 ng/ml) were observed 1 day after transfer of red drum to a black-background tank without illumination. Fish exposed to black backgrounds had the darkest body coloration, the greatest dispersion of melanin in their melanophore cells, and the most numerous and active alpha-melanophore-stimulating hormone (alpha-MSH) cells. These results provide evidence that SL may be involved in adaptation of red drum to different backgrounds and illuminations. However, the relative importance of SL and alpha MSH in the regulation of this process remains to be elucidated. PMID- 8713642 TI - Reduction in penis size and plasma testosterone concentrations in juvenile alligators living in a contaminated environment. AB - The development of the male reproductive ducts and external genitalia in vertebrates is dependent on elevated androgen concentrations during embryonic development and the period of postnatal growth. We have observed that a population of juvenile alligators living on Lake Apopka exhibit significantly smaller penis size (24% average decrease) and lower plasma concentrations of testosterone (70% lower concentrations) when compared to animals of similar size on Lake Woodruff. In addition to smaller phalli, no relationship exists between plasma testosterone concentrations and penile size in males from Lake Apopka, whereas a positive relationship exists for males from Lake Woodruff. The alligators on Lake Apopka are known to have elevated concentrations of the antiandrogenic DDT breakdown product p.p'-DDE stored in their fat. We suggest a number of hypotheses that could explain the modification in the phenotype of the juvenile male living in Lake Apopka. These modifications in phenotype include a smaller penis size, lower plasma androgen concentrations, and lack of responsiveness of the penis to the plasma androgens present. PMID- 8713643 TI - The effects of growth hormone on avian skeletal muscle satellite cell proliferation and differentiation. AB - Growth hormone receptor (GH-R) mRNA was expressed in avian skeletal muscle tissue and satellite cells in culture, and was capable of binding chicken growth hormone (cGH). In the satellite cells, GH-R gene expression was regulated by cGH in a biphasic manner which correlated with the GH effect on cell proliferation: 2-10 ng/ml of the hormone increased GH-R mRNA and DNA synthesis, whereas higher concentrations attenuated these effects. GH induced insulin-like growth factor I (IGF-I) mRNA, a potential factor for satellite cell proliferation and differentiation. However, GH inhibited the gene expression of myogenin and the expression of muscle-specific proteins in a dose-dependent manner. These results suggest a role of GH for inhibiting satellite cell differentiation in an IGF-I independent manner. During satellite cell differentiation, both GH-R mRNA expression and cGH binding peaked when cells were still proliferating and beginning to fuse, and then declined as cells fully differentiated. GH-R mRNA expression in muscle tissue and the satellite cell fraction was evaluated during chicken growth. In both fractions, GH-R mRNA peaked at 4 days of age and then declined in correlation with the reduction of muscle regulatory gene expression. Our results are in contrast with previous studies on rat muscle satellite cells, suggesting a difference between mammalian and avian species in the mode of action of GH in these cells. Our notion is that GH, via its own receptor, promotes more satellite cells to proliferate by inhibiting their differentiation, leading to the addition of more nuclei to the growing muscle. PMID- 8713644 TI - Influence of photoperiod, time, and sex on hormone concentrations in collared lemmings (Dicrostonyx groenlandicus). AB - Collared lemmings (Dicrostonyx groenlandicus) of both sexes show seasonal changes in body mass and body composition. Previous studies using single-point sampling indicated that, in young males, these photoperiod-mediated changes are associated with changes in circulating growth hormone (GH), corticosterone (B), and thyroid hormones. The present study was conducted to (1) examine daily fluctuations in serum levels of GH, B, and thyroxine (T4) in animals exposed to long (22L:2D, "LD"), intermediate (16L:8D, "ID"), and short (8L:16D, "SD") photoperiods, (2) confirm that conclusions based on single-point sampling are valid when photoperiod-related differences in hormone concentration are examined over 24 hr, (3) examine the effect of photoperiod on hormone concentrations in adults of both sexes, and (4) characterize the daily pineal melatonin rhythm in this species. Adult male and female collared lemmings housed in SD had higher levels of GH, and lower levels of B and T4, even when the diurnal variations in serum concentrations of these hormones were taken into account. A significant effect of time was observed on serum B (ID animals only) and serum T4. ID lemmings had B levels that were similar to those of SD animals, but concentrations of GH that more closely resembled those of LD animals. Females had lower GH and T4 than males. Pineal melatonin concentration closely tracked the dark phase of the day in each of the three photoperiods. Photoperiod-mediated changes in melatonin synthesis may mediate observed day length-related differences in serum concentrations of metabolic hormones, which in turn may contribute to the seasonal changes in body composition observed in collared lemmings. PMID- 8713645 TI - Radioligand and immunochemical studies of turtle oviduct progesterone and estrogen receptors: correlations with hormone treatment and oviduct contractility. AB - Progesterone (PR) and estrogen (ER) receptors were previously identified and characterized in the reproductive tract of the turtle, Chrysemys picta, and changes in PR levels were monitored during the seasonal cycle. To understand the hormonal regulation of PR, intact and ovariectomized animals were treated with estradiol, progesterone, and a combination of estradiol and progesterone, and high affinity PR and ER levels were determined by radioligand binding studies. Ovariectomy significantly decreased ER levels; in contrast, PR levels increased following ovariectomy. In both intact and ovariectomized animals, estradiol alone did not elevate PR levels above control; however, the PR was down-regulated by progesterone. ER levels in ovariectomized animals were not restored by any of the steroid regimens. By Western blot analysis, PR levels appeared to increase following ovariectomy, were unaffected by estradiol, and were somewhat decreased following progesterone treatment in estradiol-primed ovariectomized animals. While not quantitative, these results are supportive of radioligand binding studies. Immunocytochemical studies of oviduct PR followed the same pattern showing increased immunoreactivity following ovariectomy, no change with estradiol, and a decrease following progesterone treatment of estradiol-primed animals. Oviduct contractility was monitored as a physiological index of progesterone action. Estradiol significantly increased the amplitude of the contractions both in vivo and in vitro, whereas progesterone in combination with estradiol significantly inhibited the estrogen effect. This study suggests that estradiol alone may not be adequate for regulation of both ER and PR. While progesterone down-regulates its own receptor, it does not appear to influence the ER. These data are in contrast to mammalian and avian studies which show that estradiol increases both the ER and PR in the reproductive tract, and progesterone down-regulates both receptors. PMID- 8713646 TI - Immunocytochemical localization of androgen receptor in the comb, uropygial gland, testis, and epididymis in the domestic chicken. AB - Nuclear, but not cytoplasmic androgen receptors (AR), were localized immunocytochemically in the comb, uropygial (preen) gland, testis, and epididymis of juvenile and adult cockerels. Androgen receptor immunoreactivity (AR-ir) was seen in the comb, in the stratum germinativum of the epidermis and in fibromucoid cells in the dermis in juvenile and adult cockerels. AR-ir was observed in the glandular epithelial (sebum-producing) cells lining the peripheral and middle sections of the tubules in the uropygial gland. AR-ir was not detected in innermost part of the tubules which conduct the sebum to the surface of the skin. AR-ir labeling was not observed in the uropygial gland of juveniles. In the testis, AR-ir was seen in the Leydig cells, in adults but not juveniles. The epithelial cells lining the tubules in the epididymis contained AR-ir in both juveniles and adults. PMID- 8713647 TI - Immunochemical detection of parathyroid hormone-related protein in the saccus vasculosus of a teleost fish. AB - Using antisera to regions of human parathyroid hormone-related protein (PTHrP) the saccus vasculosus (SV) of the sea bream (Sparus aurata) has been shown to contain immunoreactive PTHrP. By immunohistochemistry (IHC) the epithelial coronet cells in fixed and wax-embedded SV tissue reacted with antisera to the prepro region of human PTHrP (-13 to +2), the N-terminus PTHrP (1-16), and the midmolecule PTHrP (50-69). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of saccus extracts and incubation media contained two major proteins of 14.3 and 15 kDa. By Western blotting these two proteins both reacted with the three antisera used for IHC, suggesting that they are immunochemically similar to human PTHrP (1-84). Ultrastructurally the coronet cells of Sparus saccus vasculosus resembled coronet cells described for other teleosts, with an abundant smooth endoplasmic reticulum (SER) which was more highly organized in the coronets. IHC at EM level showed reaction mainly with the membranes of the SER. These results suggest that S. aurata saccus vasculosus may produce a PTHrP like molecule similar to human PTHrP. PMID- 8713648 TI - Roles of calcium and calmodulin in the mediation of acute and sustained GnRH stimulated gonadotropin secretion from dispersed goldfish pituitary cells. AB - The possible involvement of extracellular Ca2+ ([Ca2+]o) in mediating the acute gonadotropin (GtH) response to salmon gonadotropin-releasing hormone (sGnRH) and chicken gonadotropin-releasing hormone-II (cGnRH-II) in goldfish was examined using dispersed pituitary cells in perifusion. Perifusion with Ca(2+)-deficient medium reduced the GtH responses to 5-min pulses of either GnRH, indicating the participation of [Ca2+]o in acute GnRH action. Using a 10-min GnRH pulse application protocol, the dependence of the acute GtH responses to the two GnRHs on [Ca2+]o entry through voltage-sensitive Ca2+ channels (VSCC) was examined using the dihydropyridine VSCC blocker nifedipine and the cation Co2+. Treatment with nifedipine consistently reduced the acute GtH response to either sGnRH or cGnRH-II. Similarly, perifusion with CoCl2 reduced the sGnRH-induced GtH release. In contrast to its effects on sGnRH, CoCl2 abolished the cGnRH-II-induced GtH release. These results indicate that [Ca2+]o entry through VSCC participates in the acute GtH response to both native GnRHs; however, the cGnRH-II-stimulated acute release is relatively more dependent on [Ca2+]o and VSCC functions than sGnRH-induced secretion. The involvement of calmodulin (CaM) in mediating GnRH action was also examined. Treatment with a CaM antagonist, calmidazolium, or with a Ca2+/CaM-dependent protein kinase II inhibitor, KN62, reduced GtH responses to sGnRH and cGnRH-II in 2-hr static incubation, but not in perifusion studies with dispersed goldfish pituitary cells using 5- or 10-min GnRH pulses. These results suggest that CaM-dependent mechanisms participate in mediating the long-term, but not the acute, GtH response to GnRH. Compared to sGnRH, cGnRH-II-induced GtH release was more sensitive to inhibition by KN62, indicating a higher degree of dependence of cGnRH-II action on CaM. These results extend our understanding of the differential involvement of [Ca2+]o and CaM in mediating the short-term and long-term actions of the two native GnRH peptides on GtH release in goldfish. PMID- 8713649 TI - Estrogen receptors in the stingray (Dasyatis akajei) ultimobranchial gland. AB - On the basis of our previous observation that estrogen stimulates calcitonin secretion by the ultimobranchial gland in the stingray, Dasyatis akajei, experiments were conducted to examine the presence of estrogen receptor (ER) and its mRNA in the gland, employing exchange assay and Northern blot analysis, respectively. The optimal incubation conditions of the exchange assay for cytosolic ER were found to be 25 degrees for 2 hr. Scatchard analysis of cytosolic ER revealed two components with dissociation constants (Kd) of 0.090 +/ 0.015 and 17.780 +/- 1.910 nM and with a maximum number of binding sites (Bmax) of 62.2 +/- 7.9 and 2290 +/- 180 fmol/mg protein, respectively. Furthermore, Sephadex G-100 filtration of the cytosol and immunoblot analysis using antiserum against a recombinant rat ER confirmed the presence of two specific binding components for estrogen (41 and 57 kDa). Total RNA extracted from the ultimobranchial gland was subjected to Northern blot analysis using a rat ER cDNA as a probe. Two positive signals were detected at 2.5 and 2.0 kb. Thus, the presence of ER was confirmed for the first time in nonmammalian calcitonin secreting cells. PMID- 8713650 TI - The effect of ovariectomy on concentrations of plasma prolactin and LH and parental behavior in the domestic fowl. AB - The role of ovarian hormones in the expression of parental behavior and in the regulation of LH secretion was investigated in incubating commercial meat-type hens. After ovariectomy, incubating hens continued to incubate eggs normally and brooded day-old chicks given to replace eggs, in a manner similar to sham ovariectomized control hens. The concentration of plasma LH increased significantly in incubating hens after ovariectomy while the concentration of plasma prolactin remained high. Plasma LH remained depressed in sham ovariectomized incubating control hens. The increase in plasma LH in incubating hens after ovariectomy (3.92 +/- 0.7 ng/ml) was less than that following the ovariectomy of nonincubating, nonlaying hens (5.3 +/- 1.2 ng/ml). The two groups of hens differed in that plasma prolactin concentrations were high (527 +/- 7.4 ng/ml) in the incubating hens and low (70 +/- 9 ng/ml) in the nonincubating hens. Nest deprivation resulted in an increase in plasma LH in both ovariectomized and sham-ovariectomized incubating hens with a significantly larger increase occurring in the ovariectomized hens (8.5 +/- 1.41 ng/ml compared to 2.48 +/- 0.65 ng/ml). Nest deprivation resulted in a similar rapid decrease in plasma prolactin in both ovariectomized and sham-ovariectomized hens. Replacement of eggs with day-old chicks in ovariectomized or sham-ovariectomized incubating hens resulted in a rapid decrease in plasma prolactin and after 6 days, in an increase in plasma LH in the ovariectomized but not sham-operated hens. It is concluded that once incubation behavior is established, ovarian hormones are not required for its maintenance or the readiness to brood day-old chicks. Ovarian hormones do, however, suppress LH release during incubation while the high concentration of plasma prolactin supplements this suppression. PMID- 8713651 TI - Developmental transitions in the myosin heavy chain phenotype of human respiratory muscle. AB - We studied the expression of myosin heavy chain (MHC) isoforms in the costal diaphragm (DIA) and the genioglossus (GG) muscles from 16 to 42 weeks gestation in the human using Western blotting techniques. Embryonic/neonatal MHC (MHCemb/neo) was the predominant isoform expressed in the DIA and GG at 16-24 weeks gestation. Subsequently, MHCemb/neo expression declined and the expression of MHCslow and MHC2A increased. At term, the DIA MHC phenotype was a composite of MHCemb/neo (15% of the total MHC complement), MHCslow (32%), MHC2A (47%), and MHC2B (6%); whereas, the GG was largely comprised of MHC2A (74%). We conclude that human DIA and GG demonstrate temporally dependent changes in MHC expression during gestation- and muscle-specific MHC phenotypes as they approach term. PMID- 8713652 TI - Prolactin and growth hormone in perinatal asphyxia. AB - To assess the pituitary response to perinatal asphyxia, the prolactin (PRL) and growth hormone (hGH) serum concentrations were measured in 55 asphyctic (15 preterm and 40 full-term) and 35 control (15 preterm and 20 full-term) newborns at 2-4, 24, 48 and 98 h of life. At 2-4 h the median PRL in the preterm asphyctic neonates was 5.2 U/l, whereas in the preterm control newborns it was 3.2. In the full-term newborns with and without hypoxic-ischaemic encephalopathy (HIE) it was 5.8 and 3.4, respectively. In the full-term neonates with HIE the PRL remained significantly higher than in the controls up to the fourth day. At 2-4 h the median hGH in the preterm asphyctic and control newborns was 106.2 and 54.8 mU/l, respectively. In full-term newborns it was 85.0 and 40.8, respectively. The lowest hGH concentrations were found in the severe HIE. The increased PRL and hGH in asphyxia may result from a stress-related hormone release, whereas the very low hGH concentrations in severe HIE may result from a damage at the hypothalamic hypophyseal axis. PMID- 8713653 TI - Differential role of endothelium-derived nitric oxide in the modulation of the systemic and pulmonary circulation in lambs. AB - We have studied the differential role of endothelium-derived nitric oxide (EDNO) in the regulation of the systemic and pulmonary circulations of the lamb. Hemodynamic effects of NG-nitro-L-arginine methyl ester (L-NAME, 1 mg/kg i.v.), an inhibitor of NO synthesis, were determined in juvenile (6 +/- 1 weeks old) lambs, under conditions of basal and elevated vasomotor tone. Under basal conditions, L-NAME raised both systemic (SVR) and pulmonary vascular resistances (PVR) by 20-30% (increasing SVR from 0.318 +/- 0.013 to 0.385 +/- 0.015 mm Hg.min.ml-1.kg and PVR from 0.050 +/- 0.003 to 0.067 +/- 0.010 mm Hg.min.ml 1.kg). When tone was elevated in the pulmonary circulation with hypoxia (PVR was elevated by 60%, from 0.059 +/- 0.010 to 0.094 +/- 0.019 mm Hg.min.ml-1.kg), L NAME treatment resulted in an augmented increase in PVR (PVR increased by greater than 50% to 0.140 +/- 0.024 mm Hg.min.ml-1.kg). However, when tone was elevated to a comparable degree in the systemic circulation with angiotensin infusion (SVR was elevated by 60%, from 0.432 +/- 0.065 to 0.065 to 0.634 +/- 0.113 mm Hg.min.ml-1.kg), the response to L-NAME was not augmented. Our data suggest that the role of EDNO in the modulation of the pulmonary circulation is dependent on the level of vasomotor tone, whereas its role in the systemic circulation is small and is independent of the level of vasomotor tone. PMID- 8713654 TI - Lung hypoplasia and surfactant system immaturity induced in the fetal rat by prenatal exposure to nitrofen. AB - We studied the biochemical maturity of the lungs of fetuses born to rats exposed to nitrofen on day 9.5 of gestation. In comparison with controls, nitrofen treated fetuses had pulmonary hypoplasia (decreased lung/body weight), lung hypocellularity (low DNA content) and cellular atrophy (low protein/DNA and phospholipid/DNA) on gestational days 19 and 21. Treated animals with congenital diaphragmatic hernia (CDH) also had cell atrophy and surfactant immaturity (decreased disaturated phosphatidylcholine/DNA) near term. Our data demonstrate that nitrofen causes lung hypoplasia and some degree of surfactant system immaturity that is particularly prominent in fetuses with CDH. PMID- 8713656 TI - The development of preatherosclerotic coronary artery lesions in perinatal piglets. AB - The fine structure and ultrastructure of the anterior descending coronary artery were studied in a series of perinatal piglets at 1 week prior to birth, and at 8, 24, 72 and 168 h after birth. In the anterior descending coronary artery, at or just distal to the branch point of the left circumflex artery, early plaque-like intimal lesions were present in the majority of animals. These consisted of subendothelial edema, fragmentation and dissolution of the internal elastic lamella, and the appearance of intimal myoid cells known as modified smooth muscle cells (MSMCs). These changes were present in all piglets at and before 8 h of age. They persisted and progressed during the first week of life in about half of the piglets. Beginning at 72 h and continuing through 168 h, there was an increase in MSMCs and the appearance of fibroblasts. Both fibroblasts and MSMCs were associated with the elaboration of dense collagen fibrils. Foamy macrophages appeared within the subendothelial intima having the appearance of lipophages. While the prevalence of these changes at birth indicates that they may be part of normal development, their persistence in half the piglets and structural features suggest reaction to intimal injury beginning prenatally. The lesions may be precursive of coronary atherosclerosis later in life and may parallel the early stages of atherogenesis in humans. PMID- 8713655 TI - Effects of the cyclooxygenase inhibitor ibuprofen on retinal and choroidal blood flow during hyperoxia in newborn piglets. AB - PURPOSE: The effect of the cyclooxygenase inhibitor ibuprofen (IB) on choroidal (ChBF) and retinal (RBF) blood flow during hyperoxia was examined in 21 spontaneously breathing newborn piglets. METHODS: ChBF and RBF were measured using radiolabelled microspheres before and 30 min after either saline or IB (30 mg/kg, i.v.) infusion in room air and subsequently, after 90 min of hyperoxia (O2). RESULTS: The basal RBF and ChBF did not change after IB infusion. However, during hyperoxia a significant decrease in RBF was observed in the IB group (54 +/- 5 to 37 +/- 3 ml/min/100 g, p < 0.02) and in the control group (54 +/- 3 to 37 +/- 5 ml/min/100 g, p < 0.02). Also, ChBF decreased in the IB group (2,635 +/- 446 to 1,670 +/- 199 ml/min/100 g, p < 0.02) and in the control group, (2,997 +/- 346 to 2,014 +/- 246 ml/min/100 g,p < 0.02) during hyperoxia. CONCLUSIONS: Despite cyclooxygenase inhibition with IB, RBF and ChBF decreased to the same extent as in the control group following exposure to O2. These data suggest that hyperoxia decrease RBF and ChBF through mechanisms and/or mediators other than the cyclooxygenase by-products of arachidonic acid metabolism. PMID- 8713657 TI - Rapid tracheal infusion of surfactant versus bolus instillation in rabbits: effects on oxygenation, blood pressure and surfactant distribution. AB - Surfactant bolus instillation may be associated with a drop in blood pressure. Platelet-activating factor (PAF) has been found in surfactant preparations. The aim of this study was to evaluate rapid tracheal infusion of surfactant during 5 min as an alternative to bolus instillation and to examine whether a PAF receptor antagonist is able to prevent the decrease in blood pressure. METHODS: Surfactant deficiency was induced in 16 adult rabbits by lung lavages with saline. Six animals received a bolus of a porcine surfactant preparation (Curosurf (CS); 200 mg/kg), labeled with red microspheres to assess pulmonary distribution. In another 5 rabbits, the same amount of labelled CS was instilled by tracheal infusion within 5 min. A third group of 5 animals received 3 mg/kg body weight of the PAF antagonist WEB 2170 before CS bolus instillation. RESULTS: After CS bolus administration, mean PaO2 increased by 44.7 +/- 8.3 kPa (mean +/- SD) within 2 min and remained at this level. Mean arterial blood pressure dropped transiently by 2.3 +/- 2 kPa within 5 min. Pulmonary distribution of surfactant was even. After infusion, mean PaO2 rose by 22.4 +/- 16.3 kPa within 15 min. Blood pressure dropped by 1.8 +/- 1.1 kPa within 15 min. The distribution was extremely uneven. Blood pressure decreases also occurred after pretreatment with PAF receptor antagonist. CONCLUSION: Rapid tracheal infusion of surfactant results in poorer oxygenation, an inhomogeneous distribution and a similar decrease in blood pressure compared to the bolus instillation method. Blood pressure changes could not be prevented by a PAF receptor-specific antagonist. PMID- 8713658 TI - Deficient quantitative expression of CD45 isoforms on CD4+ and CD8+ T cell subpopulations and subsets of CD45RA(low)CD45RO(low) T cells in newborn blood. AB - Deficiencies in the quantitative expression of CD45RA and CD45RO on CD4+ and CD8+ T cells and in a population of CD45RA(low)CD45RO(low) T cells in blood from term newborn infants were found by flow cytometry. The relative frequencies of CD45RO on CD4+ T cells from adults and newborn infants were 72 and 58%, respectively. However, in newborn infants greater than 70% of T cells expressing CD45RO also expressed CD45RA. In addition, the quantitative expression of CD45RA and CD45RO on newborn T cells was significantly less than that found on adult blood T cells. PMID- 8713659 TI - A community model of ciliate Tetrahymena and bacteria E. coli: Part I. individual based models of tetrahymena and E. coli populations. AB - The dynamics of a microbial community consisting of a eucaryotic ciliate Tetrahymena pyriformis and procaryotic Escherichia coli in a batch culture is explored by employing an individual-based approach. In this portion of the article, Part I, population models are presented. Because both models are individual-based, models of individual organisms are developed prior to construction of the population models. The individual models use an energy budget method in which growth depends on energy gain from feeding and energy sinks such as maintenance and reproduction. These models are not limited by simplifying assumptions about constant yield, constant energy sinks and Monod growth kinetics as are traditional models of microbal organisms. Population models are generated from individual models by creating distinct individual types and assigning to each type the number of real individuals they represent. A population is a compilation of individual types that vary in a phase of cell cycle and physiological parameters such as filtering rate for ciliates and maximum anabolic rate for bacteria. An advantage of the developed models is that they realistically describe the growth of the individual cells feeding on resource which varies in density and composition. Part II, the core of the project, integrates models into a dynamic microbial community and provides model analysis based upon available data. PMID- 8713660 TI - A community model of ciliate Tetrahymena and bacteria E coli: Part II. interactions in a batch system. AB - Premised on relatively simple assumptions, mathematical models like those of Monod, Pirt or Droop inadequately explain the complex transient behavior of microbial populations. In particular, these models fail to explain many aspects of the dynamics of a Tetrahymena pyriformis-Escherichia coli community. In this study an alternative approach, an individual-based model, is employed to investigate the growth and interactions of Tetrahymena pyriformis and E. coli in a batch culture. Due to improved representation of physiological processes, the model provides a better agreement with experimental data of bacterial density and ciliate biomass than previous modeling studies. It predicts a much larger coexistence domain than rudimentary models, dependence of biomass dynamics on initial conditions (bacteria to ciliate biomasses ratio) and appropriate timing of minimal bacteria density. Moreover, it is found that accumulation of E. coli sized particles and E. coli toxic metabolites has a stabilizing effect on the system. PMID- 8713661 TI - A new bell-shaped function for idiotypic interactions based on cross-linking. AB - Most recent models of the immune network are based upon a phenomenological log bell-shaped interaction function. This function depends on a single parameter, the "field," which is the sum of all ligand concentrations weighted by their respective affinities. The typical behavior of these models is dominated by percolation, a phenomenon in which a local stimulus spreads globally throughout the network. The usual reason for employing a log bell-shaped interaction function is that B cells are activated by cross-linking of their surface immunoglobulin receptors. Here we formally derive a new phenomenological log bell shaped function from the chemistry of receptor cross-linking by bivalent ligand. Specifying how this new function depends on the ligand concentrations requires two fields: a binding field and a cross-linking field. When we compare the activation functions for ligand-receptor pairs with different affinities, the one field and the two-field functions differ markedly. In the case of the one-field activation function, its graph is shifted to increasingly higher concentration as the affinity decreases but keeps its width and height. In the case of the two field activation function, the graph of a low-affinity interaction is nested within the graphs of all higher-affinity interactions. We show that this difference in the relations among activation functions for different affinities radically changes the network behavior. In models that described B cell proliferation using the one-field activation function, network behavior was dominated by low-affinity interactions. Conversely, in our new model, the high affinity interactions are the most significant. As a consequence, percolation is no longer the only typical network behavior. PMID- 8713662 TI - Biometrics, biomathematics and the morphometric synthesis. AB - At the core of contemporary morphometrics--the quantitative study of biological shape variation--is a synthesis of two originally divergent methodological styles. One contributory tradition is the multivariate analysis of covariance matrices originally developed as biometrics and now dominant across a broad expanse of applied statistics. This approach, couched solely in the linear geometry of covariance structures, ignores biomathematical aspects of the original measurements. The other tributary emphasizes the direct visualization of changes in biological form. However, making objective the biological meaning of the features seen in those diagrams was always problematical; also, the representation of variation, as distinct from pairwise difference, proved infeasible. To combine these two variants of biomathematical modeling into a valid praxis for quantitative studies of biological shape was a goal earnestly sought though most of this century. That goal was finally achieved in the 1980s when techniques from mathematical statistics, multivariate biometrics, non Euclidean geometry and computer graphics were combined in a coherent new system of tools for the complete regionalized quantitative analysis of landmark points together with the biomedical images in which they are seen. In this morphometric synthesis, correspondence of landmarks (biologically labeled geometric points, like "bridge of the nose") across specimens is taken as a biomathematical primitive. The shapes of configurations of landmarks are defined as equivalence classes with respect to the Euclidean similarity group and then represented as single points in David Kendall's shape space, a Riemannian manifold with Procrustes distance as metric. All conventional multivariate strategies carry over to the study of shape variation and covariation when shapes are interpreted in the tangent space to the shape manifold at an average shape. For biomathematical interpretation of such analyses, one needs a basis for the tangent space compatible with the reality of local biotheoretical processes and explanations at many different geometric scales, and one needs graphics for visualizing average shape differences and other statistical contrasts there. Both of these needs are managed by the thin-plate spline, a deformation function that has an unusually helpful linear algebra. The spline also links the biometrics of landmarks to deformation analysis of the images from which the landmarks originally arose. This article reviews the history and principal tools of this synthesis in their biomathematical and biometrical context and demonstrates their usefulness in a study of focal neuroanatomical anomalies in schizophrenia. PMID- 8713663 TI - A model for treatment strategy in the chemotherapy of AIDS. AB - Mathematical models are developed for the chemotherapy of AIDS. The models are systems of differential equations describing the interaction of the HIV infected immune system with AZT chemotherapy. The models produce the three types of qualitative clinical behavior: an uninfected steady state, an infected steady state (latency) and a progression to AIDS state. The effect of treatment is to perturb the system from progression to AIDS back to latency. Simulation of treatment schedules is provided for the consideration of treatment regimes. The following issues of chemotherapy are addressed: (i) daily frequency of treatment, (ii) early versus late initiation of treatment and (iii) intermittent treatment with intervals of no treatment. The simulations suggest the following properties of AZT chemotherapy: (i) the daily period of treatment does not affect the outcome of the treatment, (ii) treatment should not begin until after the final decline of T cells begins (not until the T cell population falls below approximately 300 mm-3) and then, it should be administered immediately and (iii) a possible strategy for treatment which may cope with side effects and/or resistance, is to treat intermittently with chemotherapy followed by interruptions in the treatment during which either a different drug or no treatment is administered. These properties are revealed in the simulations, as the model equations incorporate AZT chemotherapy as a weakly effective treatment process. We incorporate into the model the fact that AZT treatment does not eliminate HIV, but only restrains its progress. The mathematical model, although greatly simplified as a description of an extremely complex process, offers a means to pose hypotheses concerning treatment protocols, simulate alternative strategies and guide the qualitative understanding of AIDS chemotherapy. PMID- 8713664 TI - [Hepatitis C virus antibody in the serum of blood donors]. AB - Authors investigated the presence of antibody to hepatitis C virus (anti-HCV) at 67,534 blood donations, and alanin-amino-transferase (ALT) levels were analysed at the same time. The prevalence of anti-HCV antibody was 0.73% at 15,864 blood donors. Frequency of positive reactions of anti-HCV antibody increased paralelly with advance of age. Increased or high serum ALT level was found at more than 50% of samples with positive HCV reactivity. At the base of follow-up the blood donors were screened out from blood donation temporarily or definitively. Anti HCV donors were taken to hepatic care. The treatment with interferon of HCV seropositive donors proved by PCR (polimerase chain reaction) was started. Transfusion and tattou are underlined as risk factors of HCV infection. The data were analysed according to sex and ages. PMID- 8713665 TI - [Six cases of basal cell nevus carcinoma in three families]. AB - Six cases of three families had basal cell nevus cacinoma syndrome of autosomal dominant inheritance. Five characteristics of this genetic disease are stressed: (1) 40% of cases had sporadic occurrence due to de novo mutations; (2) there are three phases in the manifestation of the disease: congenital abnormalities diagnosed after birth; nevoid phase during childhood with increase at adolescence; oncogen phase after the second decade; (3) symptoms have a variability and age-dependency, (4) this mutant gene can cause both congenital abnormalities and tumours; (5) these patients are very sensitive for environmental mutagens thus it is necessary to limit or to exclude UV and X-rays, cytostatic and immunosuppressive drug treatments. PMID- 8713666 TI - [Correlations between clinical picture and coronary angiography in unstable angina]. AB - In a one year period (from 01.07. 1993 to 30. 06. 1994) 103 patients were admitted to the Central Intensive Care Unit of the Albert Szent-Gyorgyi Medical University with the diagnosis of unstable angina. In this cohort of patients the authors assessed the correlation of clinical and angiographic data. Significant coronary artery disease was found in 84% (85 patients), single-vessel disease in 23% (24 patients), double-vessel disease in 38% (29 patients), triple-vessel disease in 20% (21 patients), left main stenosis in 8% (8 patients). The culprit lesion was determined in 73 cases. The distribution of the culprit lesion severity was the following: 50-70% in 17% (12 cases), 70-90% in 27% (20 cases), greater than 90% in 44% (32 cases), 100% in 12% (9 cases). Simplex lesions were seen in 43 cases, complex lesions in 9 cases, diffuse irregularities in 5 cases and total occlusions in 9 cases. Abnormalities indicating intracoronary thrombin us were seen on 5 coronarograms. No correlation could be demonstrated between the clinical classes according to Braunwald and the angiographic morphology. PMID- 8713667 TI - [Fluorescence in situ hybridization in prenatal diagnosis. First experiences]. AB - The authors examined the potential use of non-radioactive in situ hybridization in prenatal screening. Probes for chromosomes 18, 13/21, 21 and X were applied on fourteen samples of peripheral lymphocytes and nine samples of chorionic villi. The aim of the study was to compare the analyzability of the two different DNA probes for chromosome 21 on six samples of chorionic villi. Six of the nine samples of chorionic villi were hybridized with probes specific for chromosome 21 and all six cases were diagnosed properly. However, we need more data to establish a screening protocol for routine prenatal cytogenetics. PMID- 8713668 TI - [Thrombosis in the small branches of the superior mesenteric artery in the early stages of essential thrombocythemia]. AB - A tumour in the large bowel in a 67-year-old female patient was observed for the first time in 1991, and that is why a hemicolectomy was performed. Microscopic examination showed mucosal ulceration with chronic inflammation, but no malignancy was found. Small arteries in the bowel wall were occluded by organized and recanalized trombi. The pathologist thought it could be a rare visceral form of Buerger's disease. Two and a half years later essential thrombocythemia was diagnosed in the patient. On investigating her case-history the authors found that the myeloproliferative disease had already been in its early clinical stage in 1991. So the thrombotic events of the small arteries can be considered as the first sign of essential thrombocythemia. The authors concluded the observation of unusual and unexplained thrombotic events may suggest myeleproliferative disorders, even if their diagnostical criteria are not yet well manifested. PMID- 8713669 TI - [Friedrich Hoffmann and Georg Ernst Stahl, classic representatives of baroque medicine]. PMID- 8713670 TI - [Hippokrates: the "sacred disease" (morbus sacer)]. PMID- 8713671 TI - Action of cholinesters on sensory nerve endings in skin and muscle. AB - 1. This is a review of the literature on the subject of the effects of cholinesters and their agonists on sensory nerve endings. 2. The present-day view is that acetylcholine (ACh) has an excitatory action on some cutaneous receptors. Responses appear to be limited to receptors served by small myelinated and un myelinated axons where responsiveness is multimodal; that is, the receptors are activated by noxious thermal and mechanical stimulation. 3. The possible role played by acetylcholine in sensory transduction processes is discussed, as are other explanations for the presence of nicotinic cholinergic receptors on the terminals of cutaneous receptors. 4. The excitatory action of ACh and succinylcholine (SCh) on muscle spindles is described. Two possible mechanisms are considered: a direct depolarizing action on the nerve terminals and indirect excitation, brought about by a contracture of the intrafusal fibres on which the sensory endings lie. 5. The technique of using SCh in combination with fusimotor stimulation is described. This has provided new information about the internal workings of muscle spindles. Brief mention is also made of the action of SCh on tendon organs and joint receptors. 6. It is concluded that a direct action by cholinesters is restricted to receptors served by small axons with multimodal functions. The precise role of such an action remains the subject of speculation. Possible clinical significance is discussed. PMID- 8713672 TI - Vascular remodelling and molecular biology: new concepts and therapeutic possibilities. AB - 1. Over the past decade major advances in molecular cell biology have greatly increased our understanding of the way in which many growth factor genes are expressed and regulated. This knowledge is currently being translated into investigations of the cardiovascular system. 2. Two growth factor families appear to play particularly important roles, the fibroblast growth factors and the transforming growth factors-beta. These are multifunctional growth factors capable of remodelling the vasculature through their effects on cell migration, proliferation and matrix formation. 3. An understanding of their regulation, properties and nature of their receptors is providing novel insights into the physiology and pathobiology of the vasculature. It is also providing highly specific targets for future therapy. PMID- 8713673 TI - Corticosteroids in essential hypertension: multiple candidate loci and phenotypic variation. AB - 1. The role of genetically determined changes in adrenal steroid production, metabolism and action in the pathogenesis of cardiovascular disease in man is considered by studying three loci that are important in corticosteroid function. 2. Variation at the glucocorticoid receptor locus can be identified as a biallelic restriction fragment length polymorphism (Bcl1); subjects with contrasting genotypes show altered skin vasoconstrictor responses to topically applied budesonide without any significant change in leucocyte receptor binding characteristics. 3. In a case control study of patients with essential hypertension, we have shown evidence of reduced 11 beta-hydroxysteroid dehydrogenase activity, with an elevated ratio of cortisol to cortisone metabolites in urine. 4. The genes encoding 11 beta-hydroxylase and aldosterone synthase are highly homologous. Studies in the Milan hypertensive rat show variation at this locus, which may account for the increased steroid synthesis noted in the hypertensive strain; in man, a chimaeric gene comprising 5' regulatory regions from 11 beta-hydroxylase and 3' coding sequence from aldosterone synthase accounts for the autosomal dominant condition Dexamethasone Suppressible Hyperaldosteronism. Variation in the precise location of the crossover site between the two genes does not account for the observed phenotypic heterogeneity in this condition. 5. Measurement of basal plasma steroid levels in subjects with essential hypertension show an increased ratio of 11 deoxycortisol/cortisol, consistent with reduced activity of 11 beta-hydroxylase in the zona fasciculata. 6. In summary, three loci involved in corticosteroid synthesis, metabolism and action can independently affect cardiovascular phenotypes; their roles in determining pathophysiological changes, including hypertension, remain to be studied. PMID- 8713674 TI - Sodium status, corticosteroid metabolism and blood pressure in normal human subjects and in a patient with abnormal salt appetite. AB - 1. A patient with severe hypertension was found to have mildly impaired 11 beta hydroxysteroid dehydrogenase (11 beta-HSD) activity on the basis of urinary steroid metabolite ratios, low plasma aldosterone, angiotensin II and renin levels and marginally low levels of plasma potassium. 2. The patient also had a compulsively high salt intake. 3. We tested the hypothesis that high salt intake may affect 11 beta-HSD activity. 4. High salt intake in normal subjects did not significantly alter either blood pressure or 11 beta-HSD activity. 5. We suggest that the potentially small hypertensive effect of the partial enzyme deficiency in our patient, also reported in patients with essential hypertension, has been markedly amplified by the very high salt intake. PMID- 8713675 TI - Endothelins contract guinea-pig pulmonary artery and enhance its adrenergic response via ET(A) receptors. AB - 1. This study has pharmacologically characterized endothelin (ET) receptor subtype(s) mediating contraction and enhancement of adrenergic contraction in guinea-pig pulmonary artery. Isometric tension of the isolated endothelium denuded ring preparations was measured in the presence of indomethacin (10(-5) mol/L) and N(G)-nitro-L-arginine methyl ester (L-NAME; 3 x 10(4) mol/L) to exclude a mechanism via endothelium, cyclo-oxygenase-generated eicosanoids and nitric oxide. 2. In the additional presence of tetrodotoxin (TTX; 3 x 10(-7) mol/L), ET-1 (10(-11)-10(-7) mol/L) concentration-dependently contracted the preparations. The rank order of potency to contract the preparations among ET receptor agonists was ET-1, sarafotoxin (STX)6b > ET-3 > IRL 1620, STX 6c. BQ-123 (7 x 10(-7)-7 x 10(-6) mol/L) concentrations-dependently shifted the concentration-contraction curve for ET-1 to the right in a parallel manner. Pretreatment with STX 6c (3 x 10(-7) mol/L for 30 min) did not significantly desensitize contractions to ET-1, ET-3 or IRL 1620 (P > 0.05; t-test, 10 d.f). 3. ET-1 (10(-10)-10(-9) mol/L) and STX 6b (10(-9)-10(-8) mol/L) significantly enhanced the electrical field stimulation-induced contraction in a BQ-123 sensitive manner (P < 0.05: t-test, 24-38 d.f), while ET-3 (10(-11)-10(-8) mol/L) and STX 6c (10(-11)-10(-7) mol/L) did not affect contractions. ET-1 (10(-11) mol/L) significantly enhanced contractions to exogenous noradrenaline in the presence of TTX (3 x 10(-7) mol/L) (P < 0.05; t-test, 16 d.f.). 4. These data indicate that the BQ-123-sensitive ET(A) receptor mediates both contraction and enhancement of adrenergic contractions in the guinea-pig pulmonary artery. PMID- 8713676 TI - Interstitial adenosine and function in rat heart in vivo: effects of adrenaline and 8-cyclopentyl-1,3-dimethylxanthine. AB - 1. Left ventricular interstitial adenosine and cardiac function were studied in open chest rats during adrenaline stimulation and P1-purinoceptor antagonism with 8-cyclopentyl-1,3-dimethylxanthine (8-CPT). 2. Cardiac microdialysate adenosine concentration was 0.10 +/- 0.01 mumol/L (n = 24) under basal conditions, giving an estimated interstitial adenosine concentration of 0.27 mumol/L. Stimulation with 3.2 and 8.0 micrograms/kg per min adrenaline increased the rate-pressure product (heart rate x systolic blood pressure) by 72 and 157%, respectively, and increased dialysate adenosine to 0.26 +/- 0.04 and 0.65 +/- 0.11 mumol/L (n = 12), respectively (interstitial concentrations of approximately 0.70 and 1.76 mumol/L). 3. Treatment with 60 micrograms/kg per min 8-CPT did not alter basal adenosine concentrations, but potentiated elevations in dialysate adenosine during infusion of 3.2 and 8.0 micrograms/kg per min adrenaline to 0.54 +/- 0.10 and 1.30 +/- 0.22 mumol/L, respectively (n = 12). Basal function and the response to 8.0 micrograms/kg per min adrenaline were unaltered by 8-CPT, whereas elevations in heart rate and rate-pressure product during stimulation with 3.2 micrograms/kg per min adrenaline were enhanced by 8-CPT (by up to 30%). 4. Studies in isolated hearts confirmed the inhibitory potency of 8-CPT at A1 vs A2 P1-purinoceptors (e.g. pK(B) of 7.7 +/- 0.2 and 6.4 +/- 0.1 for 5'-N-ethyl carboxamidoadenosine-mediated bradycardia and vasodilatation, respectively; n = 6). Studies in intact animals verified effective A1 blockade by 60 micrograms/kg per min 8-CPT, but also revealed some inhibition of A2-mediated responses. 5. In conclusion, the data show that cardiac interstitial adenosine levels exist within a physiologically active range in vivo and increase dose-dependently during graded adrenaline stimulation. Adenosine receptor antagonism enhances elevations in interstitial adenosine and modifies functional responses to moderate, but not high, doses of adrenaline. Whether 8-CPT-dependent elevations in interstitial adenosine are due to A1 inhibition vs inhibition of A2-mediated vasodilatation requires further investigation. PMID- 8713677 TI - Effect of 8-sulfophenyl theophylline on endogenous noradrenaline release from sympathetic nerves of the rabbit ear artery. AB - 1. The release of endogenous noradrenaline (NA) and adenyl purine (ATP, ADP, AMP and adenosine) from the rabbit ear artery, evoked by electrical stimulation (ES; 16 Hz), was examined. 2. ES evoked a significant release of NA and purine; the ratio of the amount of total purine released to NA released was approximately 180 on a molar base. 3. ES-evoked purine release was significantly reduced by the denudation of the endothelium and abolished by the alpha 1-adrenoceptor antagonist, prazosin (1 mumol/L). 4. ES-evoked NA release was significantly reduced by a P1-purinoceptor antagonist, 8-sulfophenyl theophylline (8SPT). Purine release was slightly reduced by 8SPT. 5. These results suggest that endogenous NA released by ES results in the release of a large amount of purine, which may, in turn, increase the release of NA by acting on prejunctional purinoceptors on sympathetic nerve terminals. PMID- 8713678 TI - Relationship of cigarette smoking to blood pressure and serum lipids and lipoproteins in men. AB - 1. The relationship of cigarette smoking to blood pressure and serum lipids and lipoproteins was studied in 7608 men, ranging from 40 to 59 years of age. Analyses were performed separately for non-drinkers and drinkers. 2. After adjusting age and body mass index (BMI) in non-drinkers and age, BMI and alcohol intake in drinkers in forward stepwise multiple regression analysis, there was a dose-dependent negative relationship between cigarette smoking and diastolic blood pressure (DBP) and high density lipoprotein cholesterol (HDL-C), regardless of drinking habit. There was a dose-dependent positive relationship between cigarette smoking and the ratio of total cholesterol (TC) to HDL-C (TC:HDL-C) in non-drinkers, but not in drinkers. There was a dose-dependent negative relationship between cigarette smoking and TC and a positive relationship between cigarette smoking and triglycerides (TG) in drinkers, but not in non-drinkers. 3. After matching age and BMI in non-drinkers, subjects who smoked more than 30 cigarettes/day had significantly lower mean values of systolic blood pressure (SBP; 4.3%; P < 0.05), DBP (3.0%; P < 0.01) and HDL-C (15.5%; P < 0.01) and higher mean values of TC:HDL-C (25.0%; P < 0.01), TG (46.8%; P < 0.01) and beta lipoprotein (12.0%; P < 0.01) than non-smokers. In drinkers, after matching age, BMI, and alcohol intake, subjects who smoked more than 30 cigarettes/day had significantly lower mean values of SBP (2.8%; P < 0.05), DBP (4.8%; P < 0.01), HDL-C (17.3%; P < 0.01) and TC (4.4%; P < 0.01) and higher mean values of TC:HDL C (15.4%; P < 0.01) and TG (45.1%; P < 0.01) than non-smokers. 4. Although the results are somewhat variable, the present study reveals that cigarette smoking is negatively associated with SBP and DBP and unfavourably associated with lipids and lipoproteins, regardless of drinking habit. PMID- 8713679 TI - Nitric oxide inhibition in an ovine model of heart failure. AB - 1. The role of nitric oxide (NO) in congestive heart failure was investigated by studying the acute haemodynamic, hormonal and renal effects of N(G)-monomethyl-L arginine (L-NMMA(, a nitric oxide inhibitor, given as incremental bolus doses in six sheep before (normal) and after induction of heart failure (HF) by rapid left ventricular pacing (LVoff+). 2. 6-NMMA caused significant initial dose-dependent rises in left ventricular systolic pressure, mean arterial pressure (MAP), peripheral resistance (PR) and left atrial pressure and declines in cardiac output in both normal and HF states (maximum response in 2-6 min). These responses were all but abolished when L-arginine was given concurrently with L NMMA. The dose-response curve for the L-NMMA-induced rise in MAP was shifted to the right following LVP (P < 0.05), which is consistent with previous observations of blunted NO synthase activity in HF. A subsequent decline in MAP and PR to below prebolus levels was observed 30-60 min after L-NMMA administration in the paced state. No significant hormonal or renal effects were observed. 3. In conclusion, the present study confirms the important haemodynamic role played by endogenous NO in the normal state and demonstrates a blunted pressor response to NO inhibition in this model of heart failure. PMID- 8713680 TI - Enhanced tissue polyamine content in the spontaneously hypertensive rat. AB - 1. Endogenous polyamines play a key role in mediating cellular growth and differentiation. Hypertension is associated with structural modifications of the circulatory system, a process that may be facilitated by polyamines. In this study, we examined whether there are elevated polyamine concentrations in the cardiovascular tissues of spontaneously hypertensive rats (SHR) relative to Wistar-Kyoto (WKY) rats. We also determined the chronic effect of 2% difluoromethylornithine (DFMO; a polyamine biosynthesis inhibitor) on tissue polyamines and hypertension. 2. SHR and WKY rats were treated with either 2% DFMO or drug-free drinking water; blood pressure was measured on alternate days and tissue polyamines were analysed at the end of the study. 3. We found that spermidine and spermine concentrations were markedly raised in the ventricles, resistance vessels and liver of the SHR, in comparison with corresponding tissues of WKY rats. DFMO did not affect SHR resistance vessel and liver polyamines, although spermidine in the ventricles was reduced. The blood pressure of neither SHR nor WKY rats was affected by DFMO. 4. In conclusion, this study shows for the first time a raised concentration of polyamines in the resistance vasculature of the SHR, in relation to their normotensive counterparts. The inability of DFMO to significantly reduce tissue polyamines in this study is in contrast to the effectiveness of 2% DFMO in other hypertension models, suggesting that polyamine homeostasis in this model may be stringently regulated. PMID- 8713681 TI - Angiotensin II activates pressor and depressor sites of the pontomedulla that react to glutamate. AB - 1. In cats anaesthetized with a mixture of alpha-chloralose (40 mg/kg) and urethane (400 mg/kg) and in rats anaesthetized with a mixture of alpha-chloralose (60 mg/kg) and urethane (800 mg/kg), changes in systemic arterial pressure (SAP), heart rate (HR) and sympathetic activities of vertebral (VNA) and renal (RNA) nerves were determined following the microinjection of angiotensin II (AngII; 0.16 mmol/L; 50 nL) into the pressor and depressor sites of the pontomedulla previously reacted to a microinjection of monosodium L-glutamate (Glu; 0.1 mol/L; 50 nL). Pressor sites included gigantocellular tegmental field (FTG) and dorsal medulla (DM) and rostral ventrolateral medulla (VLM). The depressor site was the caudal VLM (CVLM). The effects of losartan (1 mmol/L; 50 nL), a specific AT1 receptor non-peptide antagonist for AngII, on responses induced by AngII in the VLM, DM and CVLM were also determined. 2. In 30% of pressor sites in the FTG, 55% in the VLM and 67% in the DM and in 76% of depressor sites in the CVLM previously exposed to Glu, microinjection of AngII to the same site produced pressor or depressor responses similar to that of Glu, but smaller in magnitude, particularly in the pressor VLM. Changes in both VNA and RNA induced by AngII were also smaller than those induced by Glu, particularly RNA from DM activation. 3. In the dorsal motor nucleus of the vagus, AngII, as Glu, produced marked bradycardia, but again this was smaller in magnitude than the bradycardia produced by Glu. 4. In rats, in the DM near or around the nucleus of the solitary tract where Glu increased SAP, microinjection of AngII (0.8 mmol/L; 60 nL) produced a depressor response, while the microinjection of 1.6 mmol/L (60 nL) AngII produced a pressor response. 5. Losartan blocked the increase in SAP induced by AngII in the VLM and DM. Decreases in SAP induced by AngII in the CVLM, however, were only slightly decreased by losartan. 6. Our data suggest that a significant portion of pressor and depressor sites of the pontomedulla contain neurons responsive to both AngII and Glu. In neurons in the VLM and DM, AngII produced pressor responses that were primarily mediated through AT1 receptors, while the depressor actions of AngII in the CVLM were not mediated by AT1 receptors. PMID- 8713682 TI - Structure-activity relationships for 4-anilinoquinazolines as potent inhibitors at the ATP binding site of the epidermal growth factor receptor in vitro. AB - 1. Structure-activity relationships are described for the inhibition of the tyrosine kinase activity (phosphorylation of a fragment of phospholipase Cg1) of the epidermal growth factor receptor (EGFR) by 4-anilinoquinazolines. These compounds are competitive inhibitors at the ATP binding site. 2. The preferred side chain is anilino-, substituted at the 3-position with small lipophilic groups. The quinazoline moiety is absolutely required for activity, but substituents on the quinazoline greatly modulate potency, with electron-donating groups favoured. The most potent analogue, the 6,7-dimethoxy derivative (compound 20), has an IC50 of 29 pmol/L and a very high selectivity for the EGFR over other tyrosine kinase enzymes. 3. The present study shows that it is possible to identify small molecules that are very potent, yet highly selective, inhibitors of a single component of the growth signal transduction pathway in cells. PMID- 8713683 TI - Effect of atrial natriuretic factor on corticotrophin-releasing hormone-induced adrenocorticotrophin release in the mature ovine foetus. AB - 1. Atrial natriuretic factor (ANF) and its analogues have been shown previously to inhibit corticotrophin-releasing hormone (CRH)-induced adrenocorticotrophin (ACTH) release, both in vivo and in vitro, and it has been suggested that ANF may be a, or the, physiological ACTH-inhibitory factor. To determine whether ANF is relevant in the regulation of ACTH secretion in the ovine foetus, the present study examines the effect of ANF on CRH-stimulated ACTH release in the mature ovine foetus. 2. Five chronically cannulated foetuses, studied between 129 and 140 days of gestation (term 145-150 days), received intraarterial infusions of ovine CRH (4 micrograms/h) 120 min after the start of a sustained infusion of human ANF5-28 (10 micrograms/h) or saline (1.2 mL/h). Appropriate control experiments were performed, with foetuses receiving ANF or saline infusion only. CRH, ACTH, ANF and cortisol levels were measured by sensitive and specific radioimmunoassays. Each animal received all four treatments, with the order being randomized and 2-3 days being allowed between experiments. 3. It was found that pretreatment (120 min) with ANF5-28 (at levels devoid of significant cardiovascular actions) had no effect on mean basal or peak ACTH and cortisol levels during CRH infusion. Given the current experimental parameters, these results suggest that ANF does not acutely modulate basal or CRH-stimulated ACTH and cortisol release in the mature ovine foetus. PMID- 8713684 TI - Indomethacin damage to rat gastric mucosa is markedly dependent on luminal pH. AB - 1. There is good evidence that acid is a prerequisite for aspirin induced gastric mucosal damage; however, there is inconsistent information available for non salicylate NSAID. The present study examines the effect of gastric luminal pH on indomethacin-induced gastric mucosal damage. 2. Macroscopic gastric mucosal damage induced by indomethacin (40 mg/kg) or vehicle, administered intraduodenally to male pylorus-ligated rats (n = 5-10/group), was assessed at four different levels of luminal pH (2,4,5.5 and 7) by means of digital planimetry. 3. There was a marked difference in the extent of damage induced by indomethacin at the different luminal pH levels (P = 0.001). There was no difference between the percentage of haemorrhagic lesions at pH 2 and 4 (P > 0.05), nor between pH 5.5 and 7 (P > 0.05). However, the damage at the high levels of luminal acidity (pH 2 and 4) was strikingly different from that at pH 5.5 and 7 (P < 0.05). 4. Gastric mucosal damage induced by indomethacin, a non salicylate NSAID, is augmented by the presence of high concentration of acid in the gastric lumen. The main finding, that indomethacin injury is markedly less above pH 4, may have clinical implications in the prevention of NSAID-induced mucosal injury. PMID- 8713685 TI - Dehydroepiandrosterone does not prevent adrenocorticotrophin-induced hypertension in conscious rats. AB - 1. We tested the hypothesis that dehydroepiandrosterone (DHEA), which prevents dexamethasone-induced hypertension in rats, may block adrenocorticotrophin (ACTH) hypertension, which has been presumed due to corticosterone excess. The blood pressure and metabolic effects of DHEA (18 mg/kg per day) were examined in sham and ACTH-treated (0.5 mg/kg per day) conscious Sprague-Dawley rates (n = 20). 2. ACTH but not sham injection increased blood pressure, water intake and urine output and decreased bodyweight. 3. DHEA administration for 10 days did not alter blood pressure or metabolic effects in either sham or ACTH-treated rats. 4. DHEA, which is known to block dexamethasone-induced hypertension, did not modify ACTH induced hypertension in the rat. This suggests either that ACTH-induced hypertension is not simply a consequence of glucocorticoid activity or that the action of DHEA in dexamethasone hypertension is not through blocking the glucocorticoid receptor. PMID- 8713686 TI - Dose dependent in vivo inhibition of human colorectal cancer (LoVo) by the gastrin receptor antagonist, CI-988. AB - 1. The dose-response relationship of a new, potent and selective cholecystokinin B receptor antagonist, CI-988, on the growth of LoVo, a human colon cancer cell line, was studied in vivo. 2. We have previously shown the growth of LoVo to be inhibited both in vitro and in vivo by CI-988. 3. LoVo was grown for 10 days in nude mice that were then treated with CI-988 at 1, 5, 10, 25 and 50 mg/kg per day orally for 20 days. 4. CI-988 significantly inhibited the growth of xenografts at all concentrations except the highest concentration of 50 mg/kg per day (P < 0.00001; ANOVA). The largest inhibitory activity was caused by the 10 mg/kg per day dose, causing a 47% inhibition at day 29 (P = 0.00003: Newman-Keul's test). 5. CI-988 may be of use in inhibiting the growth of colorectal cancer. PMID- 8713687 TI - A study of angiotensin II receptors after chronic inhibition of nitric oxide synthase in the spontaneously hypertensive rat. AB - 1. Nitric oxide (NO) synthase inhibition, induces a sustained increase in blood pressure and amplifies the pressor response to infused angiotensin II (AngII). This study was designed to investigate the contribution of AngII receptors in the elevated blood pressure and enhanced pressor response to AngII in the spontaneously hypertensive rat (SHR) chronically treated with N(G)-nitro-L arginine-methyl ester (L-NAME). 2. Two groups of 13 week old female SHR were housed four to a box. Group I rats received L-NAME for 7 days (2.5 mg/kg per day) in their drinking water. Group II rats received water only. Blood pressure was monitored daily by tail-cuff plethysmography. Plasma AngII was measured by radioimmunoassay. Aortic and uterine receptor binding was determined by saturation analysis using [125I]-Sar8, Ile1)AngII. Data was analysed using the computer program LIGAND. 3. Mean systolic blood pressure was significantly elevated in rats treated with L-NAME compared with the control group. Plasma AngII concentration was slightly decreased in rats treated L-NAME compared with control. Densities of both aortic and uterine AngII receptors increased significantly following NO synthase inhibition. Receptor affinity in the aorta decreased in the L-NAME group compared with control. However, uterine AngII receptor affinity was unchanged. 4. We conclude that the increased blood pressure and enhanced pressor responsiveness that occurs with chronic inhibition of NO synthesis may result partly from increased vascular AngII receptor expression. PMID- 8713688 TI - Blood pressure lowering in patients with cerebrovascular disease: results of the PROGRESS (Perindopril Protection Against Recurrent Stroke Study) pilot phase. AB - 1. Among patients with cerebrovascular disease, there is a direct and continuous association between blood pressure and the risk of stroke, but previous trials of blood pressure lowering in this patient group have been inconclusive. 2. PROGRESS (Perindopril Protection Against Recurrent Stroke Study) is a multicentre, randomized, placebo-controlled trial that aims to determine reliably the effect of angiotensin converting enzyme (ACE) inhibitor-based blood pressure lowering on stroke risk in patients with a history of cerebrovascular disease. If a 4 week run-in period on active perindopril is well tolerated, participants are randomized to either perindopril (4 mg) +/- indapamide (2.5 mg) or matching placebo(s). The primary study outcome is stroke and follow-up is for a minimum of 4 years. 3. For the pilot study nearly 5000 medical records were screened and 60 patients with recent cerebrovascular events were approached directly in hospital or at a clinic visit. Sixty-seven patients entered the run-in phase (52 from retrospective screening and 15 by prospective approach) and 60 patients proceeded to randomization. Treatment with perindopril was well tolerated; only three patients were withdrawn due to side effects and four were withdrawn for other reasons. The mean age of randomized patients was 68 years; 70% were male and 55% were 'non-hypertensive'. The mean entry blood pressure was 142/83 mmHg and following pre-randomization treatment this was reduced by 7/4 mmHg. 4. Most patients were identified by retrospective review of medical records, but this was less efficient than prospective methods. Blood pressure lowering was well tolerated by both hypertensive and non-hypertensive patients with cerebrovascular disease. The small numbers of patients and the non-randomized nature of the data reported limit the conclusions that can be drawn, but the results confirm the feasibility of the main study. PMID- 8713689 TI - Cotrimoxazole. Rationale for re-examining its indications for use. AB - Trimethoprim was specifically developed in the late 1960s as a sulphonamide potentiator and was launched in combination with sulfamethoxazole as cotrimoxazole. Laboratory data showed synergy of antimicrobial action for the combination and suggested that the use of both agents would delay the emergence of resistance. However, the tissue distribution of trimethoprim and sulfamethoxazole does not favour synergy, and resistance among common pathogens to sulfamethoxazole is high. Clinical studies comparing trimethoprim alone with cotrimoxazole for the treatment of respiratory tract and urinary tract infections have failed to show any benefit from the combination. The development of delayed resistance by use of the combination has not been substantiated. The common adverse effects seen with cotrimoxazole are gastrointestinal disturbances and skin rashes which are well described adverse effects of sulphonamides. Comparative studies suggest that these are less common with trimethoprim alone. Serious adverse effects such as liver disorders and Stevens-Johnson syndrome appear more common with cotrimoxazole. Where there is little evidence for benefit from the use of the combination, the exposure of patients to the additional risk from the adverse effects and drug interactions of 2 drugs cannot be justified. Therefore use of cotrimoxazole should be restricted to those situations such as Pneumocystis carnii pneumonia where the combination has been shown to be beneficial. PMID- 8713690 TI - Monoamine oxidase inhibitors. An update on drug interactions. AB - After initial enthusiasm, the use of monoamine oxidase inhibitors (MAOIs) has been limited by the wide range of MAOI-drug and MAOI-food interactions that are possible, particularly with sympathomimetic medications or tyramine-containing foods, resulting in hypertensive reactions. Despite their clinical benefits, this has led to a reduction in use of such medications. Discovery of the 2 main subgroups of monoamine oxidase, types A and B, led to the synthesis of MAOIs selective for one or other of these isoenzymes. Consequently, selegiline (deprenyl), a selective MAO-B inhibitor, was developed for the treatment of idiopathic Parkinson's disease. This drug is useful in the treatment of the early stages of the disease and later on as an adjunct to other drug therapies. Although the selective MAO-A inhibitor, clorgiline (clorgyline), was found to be effective in the treatment of depression, it still retained the potential to cause hypertensive reactions. Recently, agents that are not only selective, but reversible in their inhibition of MAO-A (RIMAs) have been synthesised (e.g. moclobemide and toloxatone), and have proven antidepressant efficacy. Whilst they are less likely to induce hypertensive reactions with the concomitant administration of sympathomimetic drugs or with tyramine-rich foodstuffs, it still seems wise to advocate care in co-prescribing potentially interacting medications and to advise a degree of caution with regard to the dietary intake of foodstuffs likely to contain a high tyramine content. Although these newer drugs represent an advance in safety, their use has, as yet, only been established in the treatment of depression. RIMAs also retain a potential for adverse interaction with other drugs. Concomitant prescription of serotonin enhancing drugs should only be undertaken with caution for patients on moclobemide, toloxatone or selegiline. Coprescription of sympathomimetic drugs should also be avoided with these newer MAOIs and patients should be advised against purchasing over-the-counter preparations that may contain sympathomimetic drugs. PMID- 8713691 TI - A comparative review of the tolerability profiles of dopamine agonists in the treatment of hyperprolactinaemia and inhibition of lactation. AB - Dopamine agonists are the treatment of choice for the majority of patients with hyperprolactinaemic disorders. Although characterised by a relatively high incidence of adverse effects, most commonly gastrointestinal, cardiovascular and neurological, these are usually mild and transient, and can be minimised by starting with a low dose and gradually increasing it, or taking the drug with food or while recumbent. Bromocriptine, introduced in 1971, is the reference preparation against which newer dopamine agonists are compared. It is effective in suppressing prolactin secretion, reducing prolactinoma size and restoring gonadal function. However, up to 12% of patients cannot tolerate the drug at therapeutic dosages. Cabergoline, a long-acting dopamine agonist administered once or twice weekly, has been shown to be significantly more effective than bromocriptine in suppressing prolactin secretion in hyperprolactinaemic patients, and is better tolerated, particularly in terms of nausea and vomiting. In suppressing physiological lactation, cabergoline is at least as effective as bromocriptine, and is associated with significantly fewer rebound symptoms and adverse effects. Quinagolide is a non-ergot dopamine agonist that is administered once daily. It has similar efficacy to bromocriptine, but is probably less effective than cabergoline in hyperprolactinaemic patients; it is not licensed for suppression of lactation. It is better tolerated than twice-daily bromocriptine, but is probably inferior to cabergoline in this regard. Neither bromocriptine, cabergoline nor quinagolide has been associated with any detrimental effect on pregnancy or fetal development. However, experience with bromocriptine is far more extensive; thus, for women requiring treatment for subfertility, this drug remains the treatment of choice in most centres, with cabergoline and quinagolide as acceptable second-line drugs in bromocriptine intolerant patients. In hyperprolactinaemic men, hyperprolactinaemic women not wishing to become pregnant, and for suppression of physiological lactation, cabergoline is recommended as first-line treatment. PMID- 8713693 TI - Photosensitivity associated with antipsychotics, antidepressants and anxiolytics. AB - Photosensitivity reactions evoked by systemic agents are the result of the effects of the agent combined with subsequent exposure to light. Photosensitivity induced by exogenous parenteral agents accounts for an increasing portion of the total undesirable effects caused by environmental chemicals. The exponential increase in the number of new drugs introduced each year may be one of the factors explaining the increased number of reports describing photosensitivity induced by exogenous agents. There are many reports of photosensitivity caused by antipsychotic and antidepressant agents. Although the majority of the research was focused on the photosensitising potential of chlorpromazine, other antipsychotics and antidepressants have been shown to cause cutaneous photosensitivity. An extensive drug history must be taken whenever a patient presents with a reaction limited to, or accentuated in, light-exposed areas. It should be remembered that these reactions may present with a wide morphological spectrum ranging from sunburn-like responses to eczematous, lichenoid and even bullous lesions, resembling porphyria cutanea tarda. In order to properly diagnose photosensitivity to systemic drugs it is important to prove photosensitivity by phototesting and to rule out other causes of systemic photosensitivity such as systemic lupus erythematosus and porphyria cutanea tarda. PMID- 8713692 TI - Labour analgesia. A risk-benefit analysis. AB - The pain associated with labour can be severe. The ideal labour analgesic does not exist and systemic opioids provide little relief. Nausea, vomiting and sedation are common adverse effects of systemic opioids. Paracervical block can relieve only the pain of the first stage of labour. The duration of analgesia obtained using paracervical block is limited and repeat blocks increase the risk of direct fetal injection. Epidural analgesia effectively relieves labour pain. The insertion of an epidural catheter can provide continuous analgesia throughout labour. In addition, the catheter can be used to provide surgical anaesthesia, should operative delivery be required. Epidural local anaesthetics commonly produce maternal hypotension and motor blockade. However, opioids potentiate the effect of epidural local anaesthetics. Thus, concomitant epidural opioid injection allows the use of lower concentrations of local anaesthetics, decreasing the frequency and severity of hypotension and motor blockade. Epidural analgesia has other, potentially catastrophic, adverse effects but, with safe clinical practice, these problems are extremely rare. Intrathecal injection of opioids or local anaesthetics also effective labour analgesia. However, no single intrathecal drug or drug combination reliably provides analgesia for the duration of labour. Many clinicians use both intrathecal and epidural analgesia as a combined spinal-epidural technique. This approach provides the rapid onset of intrathecal drugs and the flexibility of continuous epidural block. Fetal heart rate decelerations occasionally follow the use of any of the above labour analgesic techniques. Most studies of the aetiology of fetal heart rate decelerations have focused on factors unique to each analgesic technique. However, the similar timing and appearance of fetal bradycardia suggests a common cause. Induction of maternal analgesia may transiently alter the balance between factors encouraging and inhibiting uterine contraction. A temporary increase in the uterotonic effects of endogenous or exogenous oxytocin may then produce a tetanic uterine contraction with subsequent decrease fetal oxygen delivery and resultant fetal bradycardia. Regardless of aetiology, these bradycardias are transient and should not produce maternal or fetal morbidity. Much controversy surrounds the effects of analgesia, especially epidural block, on the course and outcome of labour. Various studies have reported that epidural analgesia slows labour, increases the incidence of malposition of the fetal head, increases the need for forceps delivery and increases the risk of caesarean delivery. Most of the studies reporting these effects are retrospective and nonrandomised. More careful studies suggest that specific anaesthetic techniques (i.e. local anaesthetic-opioid mixtures) or obstetrical management can limit or eliminate these 'risks' of epidural labour analgesia. PMID- 8713695 TI - Inhibition of colon cancer precursors in the rat by sulindac sulphone is not dependent on inhibition of prostaglandin synthesis. AB - The non-steroidal anti-inflammatory drug, sulindac, inhibits the growth of colorectal tumours in animal models of colon cancer and causes regression of polyps in patients with familial adenomatous polyposis. The mechanism by which sulindac exerts this inhibitory effect is not known, but it has been postulated to be via the inhibition of prostaglandin synthesis. However, two recent studies have indicated that sulindac sulphone, the non-prostaglandin inhibiting metabolite of sulindac, may be important in tumour inhibition. In the present study, we examined the effect of sulindac sulphone on the formation of aberrant crypt foci, the earliest identifiable lesions in the development of colorectal cancer, in the rat colon. We have previously shown that sulindac causes a dose dependent inhibition of aberrant crypt formation in this model. Aberrant crypt foci were induced with two oral doses of 1,2-dimethyl hydrazine at 25 mg/kg per dose. Treatment with sulindac sulphone at either 10 mg/kg b.d., or 20 mg/kg, b.d., was started on the day following administration of the first carcinogen dose and was continued for 3 weeks. Colons were then removed and examined for aberrant crypt foci. Colonic crypts were visualized by staining the unsectioned colon in 0.2% methylene blue solution. There was a significant reduction in the number of aberrant foci in rats treated with sulindac sulphone at 20 mg/kg, b.d. (ANOVA, P = 0.0054). The mechanism by which non-steroidal anti-inflammatory drugs inhibit formation of aberrant crypt foci is not clear; however, these data suggest that it is not due to the inhibition of prostaglandin synthesis. PMID- 8713694 TI - Oral contraceptives and cancer. A review of the evidence. AB - The following review considers epidemiological data published from 1990 onwards on oral contraceptives (OCs) and the risk of cancers of the breast, cervix uteri, endometrium, ovary, liver and skin. In several studies, breast cancer risk was seen to be elevated among women who were current users of an OC, or had recently stopped using an OC, whereas there was no residual risk 5 or more years after stopping OC use. No interaction was observed between type of OC, or with any recognised risk factor for breast cancer, or time-factor, except for some potential excess risk for women who started OC use at a young age. Most studies have confirmed that OCs moderately increase the risk of cervical cancer, particularly in human papilloma virus (HPV)-positive women, thus suggesting that OCs may act as a promoter for HPV-induced carcinogenesis. Recent epidemiological studies have confirmed that combined OCs provide substantial protection against endometrial and ovarian cancers, and results suggest that such protection is long lasting, and may persist for 15 years or more after stopping OC use. Most case control studies have shown a relationship between OC use and hepatocellular carcinoma. However, data from cohort studies or analysis of vital statistics indicate that the public health impact of such an association is modest, if not negligible. No association was observed between combined OC use and the incidence of skin melanoma, or any other common skin neoplasm. In terms of clinical and public health implications, the most relevant points regarding OC use are: (i) recent data confirm that OCs confer presistent protection against ovarian cancer; and (ii) any increased risk of breast cancer in OC users is moderate and is restricted to current/recent users. This is reassuring for younger women, whose baseline risk of this disease is extremely low. PMID- 8713697 TI - DNA flow-cytometric analysis in colorectal cancer: a comparison of metastasizing and non-metastasizing tumours. AB - The most common cause of death in patients with colorectal cancer is metastatic liver disease. In order to identify patients at a high risk of developing hepatic secondaries from colorectal cancers, DNA content was measured in metastasizing colorectal primaries (Group I, n = 32) as well as in their subsequently resected liver secondaries and in sections of non-metastasizing colorectal cancers (Group II, n = 25). A modified interpretation system involving both a DNA index and percentage of cycling cells (those in S and G2 + M phases) was developed. DNA content was measured in paraffin-embedded sections by flow cytometry using internal controls (human peripheral blood mononuclear cells) and non-malignant tissue controls (19 patients with diverticular disease). In Group I there were significantly more tumours with both abnormal ploidy (aneuploid or abnormal tetraploid peak) and > 15% cycling cells compared with Group II (Chi-squared; P = 0.034). The combination of abnormal ploidy and > 15% cycling cells was superior to Dukes' classification for identifying metastasizing tumours (Logistic Regression; P = 0.047). However, it was not possible to discriminate between the two groups using either DNA ploidy or the percentage of cycling cells alone. The metastasizing colorectal cancers exhibited similar DNA ploidy characteristics and had a similar percentage of cycling cells compared with their liver metastases. These results suggest that tumour DNA ploidy plus the percentage of cycling cells may predict the development of liver metastases and thus survival in patients with colorectal cancer. PMID- 8713696 TI - Measurement of faecal alpha 1-antitrypsin: methodologies and clinical application. AB - Presence of the serum protein, alpha 1-antitrypsin, in the faeces is a potentially useful marker for gastrointestinal disease and/or blood loss. In an effort to simplify faecal sampling procedures, we evaluated the performance of measuring alpha 1-antitrypsin in eluants of thin smears of faeces made on filter paper, relative to conventional measurement in aqueous extracts of stool. Faecal specimens and smears were collected from healthy subjects (n = 22) and from patients with gastrointestinal bleeding (n = 12) or inflammation (n = 22), colorectal neoplasia (n = 15), or miscellaneous diseases with a low risk of excessive faecal protein loss (n = 30). alpha 1-Antitrypsin was measured by ELISA and haem porphyrins (as a measure of blood loss) by HemoQuant. Results from smears were highly correlated (r = 0.81; P < 0.001) with faecal alpha 1 antitrypsin. The smear method detected an elevated faecal alpha 1-antitrypsin with 93% specificity and 75% sensitivity. Sensitivity was high ( > 88%) where levels were markedly elevated in inflammatory and bleeding groups and low ( < 62%) where abnormal levels were mildly elevated (neoplasia and miscellaneous groups). Elevated alpha 1-antitrypsin detection by either method positively predicted > 90% of patients with gastrointestinal inflammatory disease when levels were elevated 6-fold and 1.5-fold or more, respectively. In 15 patients with colorectal neoplasia, faecal alpha 1-antitrypsin was elevated in 10 patients, haem porphyrins in nine patients and either in 12; however, smear eluant levels were elevated in only six patients. Blood loss was probably a major contributor to elevated faecal alpha 1-antitrypsin in some patients but not in the inflammatory group as a whole. The sampling and aesthetic advantages of the smear eluant method are offset by reduced sensitivity, precluding its use as a screening test for colorectal neoplasia. However, its performance in predicting inflammatory disease is equivalent to that of conventional measurement and warrants a prospective evaluation as an early investigative test. Concurrent evaluation of blood loss may improve its interpretation. PMID- 8713698 TI - Regeneration of nerve fibres across a colonic anastomosis in the guinea-pig. AB - Resection and re-anastomosis of the bowel interrupts enteric neuronal pathways. The reestablishment of neuronal connections across a colonic anastomosis was studied using immunohistochemical, retrograde tracing and physiological techniques. In control guinea-pig proximal colon, retrograde labelling with 1,1' didodecyl-3,3,3,3'-tetramethylindocarbocyanine perchlorate (DiI) revealed that enteric neurons with anally-directed projections are more numerous and have longer axons than orally-projecting neurons. In resected bowel, up to 26 weeks after re-anastomosis, descending neuronal pathways were substantially interrupted. Immunohistochemical labelling of nerve fibres revealed that some enteric nerve fibres did regenerate across narrow regions of the anastomosis, growing preferentially in the oral to anal direction. However, nerve fibres immunoreactive for neurofilament protein triplet were substantially depleted in myenteric ganglia anal to the anastomosis, even after the longest recovery periods, demonstrating that axonal regrowth was limited. This was confirmed in retrograde tracing studies, as no nerve cell bodies oral to an anastomosis were labelled when DiI was placed on myenteric ganglia just anal to the anastomosis. Physiological studies confirmed that regrowth of nerve fibres across the anastomosis occurred and that it was asymmetric, as electrical stimulation led to aboral conduction across the anastomosis more reliably than oral conduction, as measured by circular muscle contraction. After resection and re-anastomosis of the colon, the disruption of neuronal pathways in the enteric nervous system was observed, with limited and preferential re-establishment of aborally-directed long connections. PMID- 8713699 TI - Effects of leminoprazole, omeprazole and sucralfate on indomethacin-induced delayed healing of kissing gastric ulcers in rats. AB - We have examined whether or not repeated treatment with indomethacin delays the healing of kissing gastric ulcers induced in rats. The effects of leminoprazole, omeprazole and sucralfate on any delay in ulcer healing caused by indomethacin were also determined in relation to myeloperoxidase activity. Kissing gastric ulcers were induced by luminal application of an acetic acid solution. Indomethacin significantly delayed ulcer healing in a dose-dependent manner. Leminoprazole and omeprazole decreased the size and depth of ulcers, the healing of which was delayed by indomethacin, while sucralfate only decreased the ulcer depth. Histological studies showed that indomethacin inhibited tissue contraction and regeneration of the ulcerated mucosa. Leminoprazole and omeprazole prevented the inhibition of these parameters. The myeloperoxidase (MPO) activity of the ulcer portion in animals treated with indomethacin was markedly higher than in the control group. Both leminoprazole and omeprazole, but not sucralfate, significantly reduced MPO activity in contrast to the control value (in the presence of indomethacin). There was a significant relationship between the ulcerated area and myeloperoxidase activity. These results suggested that: (i) leminoprazole and omeprazole prevent the indomethacin-induced delay in ulcer healing by promoting tissue contraction and regeneration of the ulcerated mucosa; (ii) sucralfate prevents the indomethacin-induced delay in ulcer healing via the promotion of the formation of granulation tissue; and (iii) MPO activity will be useful to biochemically ensure the healing state of ulcers. PMID- 8713700 TI - Cold activation of complement in sera from patients with persistent hepatitis C virus infection on interferon therapy. AB - The loss of haemolytic activity in sera during storage at low temperature (the cold activation of complement) was observed in 136 of 184 (74%) patients with chronic liver disease associated with hepatitis C virus (HCV) infection. This was more frequent than observed in the three of 40 (8%) patients with chronic hepatitis B (P < 0.001) or none in 43 normal controls (P < 0.001). Of 103 patients with chronic hepatitis C who had completed a full course of recombinant interferon-alpha 2a therapy (total dose: 516 x 10(6) U), 40 responded completely and 21 responded partially, as judged by the normalization or decrease of alanine aminotransferase levels 6 months after the completion of therapy; 42 patients did not respond at all. The cold activation of complement persisted in five (13%) complete responders, less often than in 33 (79%) non-responders (P < 0.001). At the completion of interferon therapy, the cold activation of complement persisted in 12 of 54 patients despite the normalization of alanine aminotransferase. Spontaneous exacerbation of hepatitis occurred in seven of 12 (58%) patients with cold activation, which was more frequent than in the four of 42 patients (10%) without it (P < 0.01). The cold activation of complement disappeared along with the loss of HCV-RNA in five of six responders during the 6 month period after the completion of interferon therapy, while both cold activation and HCV-RNA persisted in all eight non-responders. These results indicate that the cold activation of complement may be useful as a marker of HCV viraemia for monitoring the response to interferon in patients with HCV infection. PMID- 8713701 TI - Increased urinary endothelin excretion in patients with liver cirrhosis. AB - The urinary endothelin level in patients with chronic liver disease was determined in order to explore its possible involvement in renal function. The plasma endothelin level was significantly higher in patients with liver cirrhosis (LC) than in those with chronic hepatitis (CH) or in control patients (C). Similarly, urinary endothelin excretion in LC was significantly increased, compared with CH and C. Urinary endothelin demonstrated a significant positive correlation with creatinine clearance. The ratio of endothelin clearance/creatinine clearance did not differ statistically among the three groups. Urinary sodium excretion in LC was positively correlated with plasma endothelin, but not with urinary endothelin. Urinary endothelin excretion demonstrated a significant negative correlation with urinary kallikrein in LC. The present data suggest that increased urinary endothelin excretion in cirrhotic patients primarily depends upon elevated plasma levels of endothelin, but not renal production. Also, a possible link between endothelin and the kallikrein kinin system in liver cirrhosis is indicated. PMID- 8713703 TI - Microheterogeneity of serum transferrin in the diagnosis of hepatocellular carcinoma. AB - Heterogeneous reactivity of human serum transferrin (Tf) with lectins was analysed using patient sera to determine whether it can be used to distinguish patients with hepatocellular carcinoma (HCC) from those with liver cirrhosis (LC). Microheterogeneity of Tf was analysed by crossed immunoaffinity electrophoresis (CIAE) with concanavalin A (Con A) and Lens culinaris agglutinin (LCA). Sample sera from 58 patients with HCC, 43 patients with LC and 10 normal controls were used in this study and the results were evaluated statistically. The increments of Con A-non-reactive (C1) and -weakly reactive (C2) species of Tf were observed in HCC compared with those of LC and Norm. Significant increase in the combined percentage of Con A- C1 + C2 species was also revealed in HCC (35.5 +/- 8.5%, mean+/-s.d.) compared with those of LC (29.1 +/- 6.8%; P < 0.001) and normal controls (17.1 +/- 2.3%; P < 0.001). The elevation of LCA-reactive (L2) species of Tf was recognized in HCC (8.2 +/- 3.8%) in comparison with those of LC (4.8 +/- 3.1%; P < 0.001) and normal controls (1.3 +/- 1.7%; P < 0.001). The increment of C1 + C2 species and/or L2 species of Tf was observed in 78% (sensitivity) of patients with HCC. The specificity, the positive predictive value and the overall accuracy were 81, 88 and 72%, respectively. Positive ratio of C1 + C2 and/or L2 species was 77 and 70% in alpha-fetoprotein low and -high producing HCC patients, respectively. These results indicate that the microheterogeneity analysis of human serum Tf is useful for distinguishing patients with HCC from those with LC and normal controls. PMID- 8713702 TI - Response to interferon in chronic hepatitis C due to mixed genotype infection. AB - We examined the response to interferon (IFN) in patients with chronic hepatitis C (CHC) due to two different genotypes of hepatitis C virus (HCV) infection. Among 64 CHC patients studied, one (2%) had HCV-RNA genotype I, 36 (56%) had genotype II, 19 (30%) had genotype III, 2 (3%) had genotype IV and 6 (9%) had both genotypes II and III. There was no significant difference in age, sex, history of blood transfusion and liver histology among patients with genotypes II, III and II + III. The HCV-RNA titre of genotype II patients was significantly higher than that of genotype III patients (P < 0.05). However, there was no significant difference in the HCV-RNA titre between genotype II + III and the other groups. The complete response rate achieved with IFN therapy was significantly higher in genotype III patients (74%) than in genotype II patients (17%; P < 0.01). Of the six patients with genotype II + III, a complete response to IFN was only achieved by two patients (33%), both of whom had a low HCV-RNA titre ( < or = 10(4.5) copies/mL) and HCV serotype 2. The remaining four patients had HCV serotype 1 and three of the patients had a high HCV-RNA titre ( > or = 10(5) copies/mL). The HCV genotype III was lost in two patients after IFN therapy. These data suggest that HCV-RNA titre and HCV serotype are important factors for predicting the efficacy of IFN therapy in patients with mixed genotype infection and show direct evidence of higher susceptibility towards CHC of patients with genotype III than genotype II. PMID- 8713704 TI - Improvement of peripheral blood lymphocyte subsets in primary biliary cirrhosis after ursodeoxycholic acid therapy. AB - Immunological abnormalities frequently observed in patients with primary biliary cirrhosis are considered to be related to the pathogenesis of this disease. We performed a prospective trial to evaluate whether immune mechanisms play a role in the effectiveness of ursodeoxycholic acid (UDCA) therapy. Fifteen female patients with primary biliary cirrhosis were followed for 1 year and were then treated with UDCA (600 mg/day) for another year. Laboratory tests, including peripheral blood lymphocyte subsets assessed by dual colour fluorescence analysis using monoclonal antibodies against respective T cell markers, were evaluated at the beginning of the study, at the start of therapy and at the end of therapy. In primary biliary cirrhosis, the proportion of cytotoxic T cells, suppressor inducer T cells and alpha beta-receptor bearing T cells were significantly lower than in healthy controls. No significant changes were observed in the proportions during the year before the therapy. These reductions, however, recovered to normal ranges after 1 year of UDCA therapy. These changes were associated with an improvement in the serum levels of aspartate aminotransferase, alkaline phosphatase, gamma-globulin and IgM. The close correlation between the improvement in the imbalance of lymphocyte subsets after the therapy and the clinical status suggests that an immunological process may play a role in the effectiveness of therapy in primary biliary cirrhosis. PMID- 8713705 TI - UDP glucuronosyltransferase in the cirrhotic rat liver. AB - In patients with cirrhosis, the elimination of drugs metabolized by glucuronidation is relatively preserved, in comparison with the metabolism of drugs by oxidation. This study explores this phenomenon at a molecular level. In cirrhotic rat livers the content of UDP-glucuronosyltransferase (UGT) was examined by immunohistochemistry and immunoblotting using three antibodies: (i) a polyclonal antibody directed against a broad number of UGT isoforms from both family 1 and family 2; (ii) a family 2-specific antibody; and a (iii) family 1 specific antibody. The steady state mRNA level of UGT of a family 2 isoform was also detected by northern blot analysis. The results demonstrate normal or increased UGT protein by immunohistochemistry and immunoblot in cirrhotic livers compared with controls. This was accompanied by increased steady state mRNA encoding the UGT isoform UGT2B1. In contrast, an isoform of cytochrome P450 (CYP2C11) was reduced markedly in both immunohistochemical staining and immunoblot analysis. These results suggest that in cirrhosis there is a comparative increase or at least a maintenance of UGT enzyme content and that this most likely occurs at a pretranslational level. PMID- 8713706 TI - Gastric mucosal energy metabolism and intracellular mucin content changes in patients with liver cirrhosis. AB - Biopsy samples from patients with liver cirrhosis were investigated for changes in gastric mucosal energy metabolism and intracellular mucin content using high performance liquid chromatography and an image analysing system. The test group consisted of eight non-cirrhotic patients with endoscopically normal mucosa (controls) and eight cirrhotic patients with oesophageal varices. The amount of ATP, energy charge level and intracellular mucin content were all significantly decreased in cirrhotic patients when compared with those of the controls. The decrease in energy charge also correlated well with the decrease in intracellular mucin content in the gastric mucosa. The results indicate that gastric mucosal energy metabolism is impaired in cirrhotic patients concomitantly with a decrease in the intracellular mucin content in the gastric mucosa. These changes may weaken defensive mechanisms against acid and NSAID, resulting in gastric mucosal injury in cirrhotic patients. PMID- 8713707 TI - Increase of sulfated ursodeoxycholic acid in the serum and urine of patients with chronic liver disease after ursodeoxycholic acid therapy. AB - The present study was undertaken in order to investigate the influence of ursodeoxycholic acid (UDCA) on the composition of sulfate-conjugated bile acids in the serum and urine of patients with chronic active hepatitis and compensated liver cirrhosis. After a 12 week UDCA treatment (600 mg/day), total serum bile acid concentration increased two-fold in patients with compensated liver cirrhosis and increased slightly in patients with chronic active hepatitis. The percentage of sulfated bile acids significantly increased in patients with both compensated liver cirrhosis and chronic active hepatitis. UDCA made up 63% of the total serum bile acids in compensated liver cirrhosis and 61% in chronic active hepatitis after UDCA treatment. Of the serum bile acids after UDCA treatment, 35.2 and 53.9% of UDCA was sulfate conjugated in compensated liver cirrhosis and chronic active hepatitis, respectively. Urinary excretion of total bile acid and UDCA after UDCA treatment in compensated liver cirrhosis were higher than in chronic active hepatitis. UDCA made up 68% of the total urinary bile acids in compensated liver cirrhosis and 64% in chronic active hepatitis after UDCA treatment. Of the urinary bile acids after UDCA treatment, 51.8 and 54.8% of UDCA was sulfate conjugated in compensated liver cirrhosis and chronic active hepatitis, respectively. UDCA treatment for compensated liver cirrhosis was less effective than for chronic active hepatitis. We found that sulfate conjugation is one of the major metabolic pathways for UDCA after UDCA treatment in chronic liver diseases. PMID- 8713708 TI - Effect of verapamil on renal haemodynamics in a portal hypertensive rat model. AB - In order to investigate the effects of verapamil on renal haemodynamics in rats with portal hypertension, verapamil was given at either a low (0.2 mg/kg) or high (2 mg/kg) dose to rats after portal vein ligation. An approximate 12% decrease in mean arterial pressure followed administration of low dose verapamil, with a significant decrease in cardiac output and renal blood flow, as well as reduced portal pressure, observed; these signs were all indicative of a rise in renal vascular resistance. In contrast, the marked fall in both mean arterial pressure and cardiac output with high dose verapamil, accompanied by a significant decrease in portal pressure and no change in renal blood flow, suggests a reduction in renal vascular resistance. This study shows that the acute effects of verapamil on renal haemodynamics may vary with the dose used. Also, acute verapamil administration tends to decrease renal blood flow to alter the autoregulation of the kidney; thus, caution should be taken in the clinical use of verapamil in the treatment of cirrhosis with portal hypertension. PMID- 8713709 TI - Current concepts of hepatic uptake, intracellular transport and biliary secretion of bile acids: physiological basis and pathophysiological changes in cholestatic liver dysfunction. AB - Hepatic sinusoidal uptake of bile acids is mediated by defined carrier proteins against unfavourable concentration and electrical gradients. Putative carrier proteins have been identified using bile acid photoaffinity labels and more recently using immunological probes, such as monoclonal antibodies. At the sinusoidal domain, proteins with molecular weights of 49 and 54 kDa have been shown to be carriers for bile acid transport. The 49 kDa protein has been associated with the Na(+)-dependent uptake of conjugated bile acids, while the 54 kDa carrier has been involved in the Na(+)-independent bile acid uptake process. Within the hepatocyte, cytosolic proteins, such as the glutathione S-transferase (also designated the Y protein), the Y binders and the fatty acid binding proteins, are able to bind bile acids and possibly facilitate their movement to the canalicular domain. At the canalicular domain a 100 kDa carrier protein has been isolated and it has been shown by several laboratories that this particular protein is concerned with canalicular bile acid transport. The system is ATP dependent and follows Michaelis-Menten kinetics. Interference with bile acid transport has been demonstrated by several chemicals. The mechanisms by which these chemicals inhibit bile acid transport may explain the apparent cholestatic properties observed in patients and experimental animals treated with these agents. Several studies have shown that Na+/K(+)-ATPase activity is markedly decreased in cholestasis induced by ethinyloestradiol, taurolithocholate and chlorpromazine. However, other types of interference have been described and the cholestatic effects may be the result of several mechanisms. Cholestasis is associated with several adaptive changes that may be responsible for the accumulation of bile acids and other cholephilic compounds in the blood of these patients. It may be speculated that the nature of these changes is to protect liver parenchymal cells from an accumulation of bile acids to toxic levels. However, more detailed quantitative experiments are necessary to answer questions with regard to the significance of these changes and the effect of various hepatobiliary disorders in modifying these mechanisms. It is expected that the mechanisms by which bile acid transport is regulated and efforts to understand the molecular basis for these processes will be among the areas of future research. PMID- 8713710 TI - Classical Kaposi's: a rare cause of gastrointestinal haemorrhage. AB - Classical Kaposi's sarcoma (KS) is an indolent neoplasm involving mucocutaneous sites predominantly in elderly Mediterranean or Jewish persons. Whilst gastrointestinal involvement is common, it is usually asymptomatic. This case report presents a case of massive gastrointestinal haemorrhage in a patient with stable cutaneous disease and outlines options for the investigation and management of this rare complication. PMID- 8713711 TI - Idiopathic adulthood ductopenia. AB - In 1981, a 26 year old man occasionally demonstrated elevated serum transaminase concentrations. He had no history of medication, or a personal or family history of jaundice, except for prolonged physiological jaundice as a neonate. Serum hepatitis B surface antigen, hepatitis C virus antibody and anti-mitochondrial antibody were absent. A wedge biopsy specimen revealed ductular proliferation, mild inflammation of the portal area and disappearance of bile ducts from 80% of the portal tracts. Serial sections demonstrated a vanishing bile duct. Endoscopic retrograde choledochopancreatography, portography and arteriography demonstrated no abnormalities. In 1994, the patient died of hepatic failure following a 12 year observation period. He was subsequently diagnosed with idiopathic adulthood ductopenia on the basis of the criteria proposed by Ludwig. PMID- 8713712 TI - Health evaluation of 1,4-dioxane. PMID- 8713713 TI - Developmental toxicity of orange B when given to rats by gavage. AB - The pyrazolone dye Orange B was given by gavage to pregnant Osborne-Mendel rats throughout gestation. Dose levels of 0, 15, 30, 100, 200, 400, or 700 mg/kg body weight were given daily. On gestation day 20, the females were killed and cesarean sections were performed. Feed consumption and maternal weight gain were not affected. No dose-related changes were seen in maternal clinical findings, implantations, fetal viability, or fetal size (weight and length). No compound related effects were seen in sternebral development. No dose-related effect was seen in the incidence of skeletal variations in fetuses or in the number of litters containing fetuses with skeletal variations. Skeletal development, as measured by the average number of ossified vertebrae, was similar in all groups. No compound-related effects were seen in soft-tissue development. PMID- 8713714 TI - Dose- (and time-) dependent blockade of pregnancy in Sprague-Dawley rats administered ammonium dinitramide in drinking water. AB - Ammonium dinitramide (ADN) is a class 1.1 explosive oxidizer that can be used in solid rocket propellant mixtures and explosives. A 90-day general toxicity/ reproductive screen performed on this compound at doses of 162, 103, 29, and 0.0 mg ADN/kg/day resulted in a treatment-related adverse effect on litter production. Incidences of animals producing litters (1/11, 3/12, 12/12, and 11/12, respectively) and mean numbers of pups per litter (7, 7, 16, and 15, respectively) both were statistically significantly less than controls. In a follow-up study, mated dams treated with ADN at the same doses and examined at gestation days (GDs) 10 and 20 showed an effect in fetus loss similar to that seen in the reproductive screen. A pre- versus postimplantation dosing regimen indicated that implantation is vulnerable to ADN effects during the preimplantation period (GDs 1-3). No implantation sites were found in the rats treated with 2000 mg ADN/L drinking water (target dose of 160 mg ADN/kg/day) during GDs 1-3. Numbers of implantation sites found in the rats treated during GDs 4-8 were similar to those found in the control group. The pituitary was not identified specifically in this study as the site of primary action, but serum progesterone was reduced by 27%, which may have resulted in reduced fertility. PMID- 8713715 TI - Reproductive and neurobehavioral effects of Sunset yellow FCF administered to mice in the diet. AB - Selected reproductive and neurobehavioral parameters were measured in mice given the color additive Sunset Yellow FCF in the diet. The additive was given at levels of 0 (control), 0.15, 0.30, and 0.60%, from five weeks of age in the F0 generation to nine weeks of age in the F1 generation. There were few adverse effects on litter size, weight, or sex ratio. Average body weight of offspring during the late lactation period was significantly increased in the low- and middle-dose groups of each sex. In the neurobehavioral parameters, swimming direction was significantly affected in a dose-related manner in male and female offspring during the early lactation period. Also in the early lactation period, surface righting and negative geotaxis were significantly affected in male offspring in the middle-dose group, and swimming head angle was significantly affected in female offspring in a dose-related manner. The dose levels of Sunset Yellow FCF in this study did produce some adverse effects in reproductive and neurobehavioral parameters. PMID- 8713716 TI - Urinary 2-thiothiazolidine-4-carboxylic acid (TTCA) as the major urinary marker of carbon disulfide vapor exposure in rats. AB - Male Sprague-Dawley rats (200-250 g; 60 per exposure group) were exposed to carbon disulfide (CS2) air concentrations of 0, 50, 150, and 500 ppm(v/v) for 6 hr/day, 5 days/week over six months. Following the exposures, nine rats from each exposure group had four sets of cumulated urines collected (between 0-8, 8-16, 16 24, and 24-48 hr). The urinary parameters measured were: 2-thiothiazolidine-4 carboxylic acid (TTCA), total thioethers (TE), and the compounds responsive to the iodine-azide (IA) test. Urinary TTCA elimination obeyed pseudo-first-order, one-compartment model kinetics of half-time (t0.5) 5.2 +/- 0.3 hr up to 16 hr of collection. The elimination of TE within 16 hr had a t0.5 of 8.5 +/- 0.6 hr. TTCA, IA, and TE were correlated highly in the first 16 hr. After 16 hr, the t0.5 for TE lengthened to 13.1 hr. At CS2 concentrations of 50, 150, and 500 ppm, the respective t0.5 for IA-responsive compounds were 12.6, 6.1, and 4.4 hr. TTCA had the highest correlation coefficient and p-value relative to CS2 exposure concentration, and also was the most sensitive, precise, and selective urinary marker. PMID- 8713717 TI - Neutral endopeptidase (NEP) and its role in pathological pulmonary change with inhalation exposure to JP-8 jet fuel. AB - Through a simulated flightline exposure protocol, Fischer 344 rats (F344) were subjected to an aerosol/vapor mix of the military jet fuel, JP-8. Previous studies with this model of lung injury have revealed significant increases in pulmonary resistance, increased alveolar clearance of 99mTcDTPA, and a decrease in bronchoalveolar lavage fluid (BALF) concentration of the neuropeptide substance P (SP). Exposures to JP-8 were nose-only and for one hour daily. Six groups of Fischer 344 rats were exposed for 7, 28, or 56 days at two JP-8 concentrations (low dose = 469-520 mg/m3/hr, high dose = 814-1263 mg/m3/hr). Exposed groups were matched with longitudinal controls. In response to JP-8 inhalation, exposure animals demonstrated a dose-dependent as well as duration determined reduction in BALF SP concentration. Both JP-8 concentrations caused significant pathological changes in lower pulmonary structures. PMID- 8713718 TI - Tissue distribution, metabolism, and excretion of 2,4,4'-trichlorobiphenyl (CB 28) in the rat. AB - The tissue distribution, metabolism, and excretion of 14C-labeled 2,4,4' trichlorobiphenyl (CB-28) were studied in the rat. Following intravenous administration, radioactivity was removed from the blood rapidly and distributed to tissues. The time-courses of varying concentrations of CB-28 in tissues and blood were described mathematically using nonlinear regression analysis. The highest concentrations of radioactivity were found in the adipose and skin, which showed distinct uptake phases. High levels were seen at 30 minutes in liver and brain, but quickly declined. Fifty-two percent of the given dose was excreted in feces within 14 days. Intraperitoneal administration to three female rats for five consecutive days provided tissues which were analyzed for metabolites. PMID- 8713719 TI - Development of the hypothalamic-pituitary-thyroid axis in humans. PMID- 8713720 TI - Regulation of fetal breathing. AB - Traditionally, the idea of transient asphyxia plus some degree of cold stimulation has been used to explain the establishment of continuous breathing at birth. This idea was nurtured by observations made in the acute fetal preparation at a time when fetal breathing was considered absent. Experimental observations made in the past two decades have challenged this traditional view. First, complete peripheral chemodenervation, essential to the hypoxic stimulus theory, did not affect fetal breathing or the establishment of continuous breathing at birth. Second, occlusion of the umbilical cord in utero, as long as some oxygenation is provided to the fetus in order to avoid fetal hypoxaemia, establishes continuous breathing in utero, in the absence of all sensorial input thought to be important for the establishment of continuous breathing. These observations led us to hypothesize the presence of a placental factor responsible for the inhibition of breathing in utero. This placental factor appears to be a peptide with a molecular mass between 3.5 and 10 kDa. This review will also explore some new observations regarding the generation of central respiratory activity in the fetus, and suggests that the rhythm generator is a neuronal network in which the unit is a pacemaker-like cell uniquely responsive to CO2. PMID- 8713722 TI - Ventricular fibrillation in the neonate: elusive or illusive? AB - Ventricular fibrillation (VF) in mature large mammals is usually a terminal event. It constitutes the most common mode of exitus and the main cause of sudden cardiac death in humans. In neonates, VF is of particular interest because it often reverse spontaneously and promptly to potentially become a non-lethal event. Accordingly, the study of VF and spontaneous defibrillation (SDF) in the neonate is key for the understanding of the developmental anatomical and functional characteristics enabling mammalian myocardium to initiate and sustain this lethal arrhythmia. Such characteristics may include age-related changes of myocardial tissue properties, mass and geometry, autonomic cardiac innervation and myocardial perfusion/hydration. In the human neonate, VF/SDF could provide a plausible framework for explaining and directing the search for the elusive evidence underlying the mechanism and age distribution of the sudden infant death syndrome. PMID- 8713721 TI - Development of the newborn GI tract and its relation to colostrum/milk intake: a review. AB - During the immediate postnatal period, the gastrointestinal (GI) tract undergoes profound growth, morphological changes and functional maturation. The oesophagus shows an accelerated cell proliferation in the epithelium and an increased production and accumulation of mucus in the glands. The stomach shows a rapid tissue growth and a marked increase in acid secretion capacity. The intestine shows increased tissue growth and marked epithelial modifications; the latter include the loss of the ability by the epithelial cells of the small intestine to absorb macromolecules, and the loss of the ability by the epithelial cells of the large intestine to synthesize digestive enzymes and to absorb amino acids and glucose. These changes are apparently related to the onset of colostrum ingestion, because starved or water-fed newborns showed little changes in the GI tract. A number of hormones and growth-promoting peptides, such as insulin, cortisol, epidermal growth factor (EGF) and insulin-like growth factor I (IGF-I), have been found at high concentrations in the maternal colostrum. There is evidence that colostrum-bone EGF and IGF-I play a role in postnatal GI development in newborns. The role of other colostrum-borne hormones and growth promoting peptides remains to be assessed. Further studies are also required to demonstrate if colostrum-borne EGF and IGF-I can be used therapeutically to those newborns with immature or diseased GI tract, such as in cases of premature birth or prenatal growth retardation or cases requiring total parenteral nutrition. PMID- 8713723 TI - Imaging of the fetal heart. PMID- 8713724 TI - Developmental changes in Ca(2+)-uptake, Na+,Ca(2+)-exchange and Ca(2+)-ATPase in freshly isolated embryonic, newborn and adult chicken heart. AB - Developmental changes in cellular Ca(2+)-transport mechanisms were studied in chick heart by determining cellular Ca(2+)-uptake and Na+,Ca(2+)-exchange activity in freshly isolated ventricular tissues of embryonic (5-18 days old), newborn (1-2 days old) and young adult (90-100 days old) heart by monitoring 45Ca influx. Ca(2+)-ATPase activity was determined in microsomal fractions at different stages of development. The Ca(2+)-uptake (per g wet tissue weight) increased with the development of embryonic as well as post-hatch chick heart, reaching a maximum in the young adult chicken. The overall increase in Ca(2+) uptake, from embryonic day 5 to young-adult stage, was more than 3 fold. The Na+,Ca(2+)-exchange activity, determined as Na(+)-gradient-induced Ca(2+)-uptake in presence of either ouabain or zero [Na+]0, showed a 6-fold increase during development of heart from the embryonic day 5 to the young adult stage. Amiloride, an inhibitor of Na+,Ca(2+)-exchange, caused a dose-dependent reduction in a ouabain-induced rise in 45Ca influx at different stages of development. The inhibitory effect of amiloride was, however, greater during later stages of development. A progressive increase in Ca(2+)-ATPase activity was also seen during development. Ca(2+)-ATPase exhibited about a 4-fold increase in activity from embryonic day 7 to the young adult. The concomitant increase in Ca(2+) uptake, Na+,Ca(2+)-exchange and Ca(2+)-ATPase activities suggests age-dependent changes in Ca(2+)-transport and storage systems of developing heart during embryogenesis and post-embryonic life. During embryogenesis the developmental increase in Na+,Ca(2+)-exchange activity was greater than that during post-hatch development of heart. However, the increase in Ca(2+)-ATPase activity was greater during post-hatch development than during embryogenesis. It is suggested that Na+,Ca(2+)-exchange and Ca(2+)-ATPase play a prominent role in maintaining cellular Ca2+ homeostasis during embryogenesis and after hatching. PMID- 8713725 TI - Respiratory responses to preoptic/anterior hypothalamic warming during sleep in kittens. AB - The preoptic/anterior hypothalamic (POAH) area of 12-48-day-old unanaesthetized, unrestrained kittens was warmed with a diathermic probe to assess respiratory responses to a central thermal challenge during sleep. During quiet sleep (QS), warming of the POAH by 1.4-3.8 degrees C induced periods of tachypnoea (panting) interspersed with periods of slower respiration; the percentage of time spent in panting increased as the kittens matured. During rapid eye movement (REM) sleep, POAH warming induced a much smaller increase in respiratory rate and no sustained panting. Analysis of the dynamics of panting (in QS only, since panting did not occur in REM) revealed several changes from breathing patterns of normal respiration. During panting, the increased respiratory rate did not result from equal changes in inspiratory and expiratory times; inspiratory times accounted for a larger portion of the decline in total respiratory cycle time. Amplitude of diaphragmatic electromyographic activity showed an age-dependent response to thermal panting, which resulted in age-dependent changes in ventilation and inspiratory drive. The interspersion of slower with faster respiratory rates suggests a competition of thermoregulatory and respiratory homeostatic mechanisms. The age-dependent ventilatory responses to thermal panting suggest greater vulnerability to thermal stress at particular ages, and may have implications for the sudden infant death syndrome. PMID- 8713726 TI - Optical imaging of the ventral medullary surface of developing kittens during ventilatory challenges. AB - We used large-array optical recording procedures to examine maturation of regional neural activity within the ventral medullary surface (VMS) of anaesthetized kittens during graded hypercapnic and hypoxic challenges. The VMS was exposed through a ventral surgical approach in 10, 20, 30, and 45-day-old kittens and in adult cats under sodium pentobarbital anaesthesia. Arterial pressure, costal diaphragmatic EMG, and ECG were continuously monitored. A coherent image conduit with 12 mu fibre resolution was attached to a charge coupled-device camera and positioned over the VMS. Reflected 660 nm light was digitized continuously at 2-s intervals during a baseline period, hyperoxic hypercapnia, (3, 5, and 10% CO2 in O2), and poikylocapnic hypoxia (6%, 9%, and 12% O2 in N2), and recovery. Sixty to seventy-five images within each epoch were averaged, and subtracted from baseline. Regional differences within the image were determined by ANOVA procedures (alpha = 0.05). During hypercapnia, an overall decrease in neural activity (increase in scattered light) occurred, which was marginally age-dependent. By 30 days, regional bidirectional reflectance changes in response to CO2 emerged in a small proportion of animals, and were similar to adult responses. Hypoxia induced a dose- and age-dependent decrease in overall scattered light. Transient "on" and "off" responses were common under both ventilatory stimuli. In 20-30-day kittens, marked rebound responses in reflectance accompanied cessation of hypoxic stimuli; such patterns were absent at other ages. At 30 days, a caudal-rostral bidirectionality in response to mild hypoxia (12% O2) began to emerge in a subset of animals. We conclude that dose dependent response to ventilatory stimuli occur in the VMS at all post-natal ages of the kitten; however, in hypoxia, the magnitude of the overall reflectance changes is diminished relative to adult patterns. Rebound responses to hypoxia are present at particular ages, and older kittens begin to show a topographical organization of neural activation. PMID- 8713727 TI - Immunocytochemical localization of renin in the developing ovine fetus. AB - The ontogeny of renin distribution in the outer cortical segments was studied by immunocytochemistry in two groups of ovine fetal kidneys; one set of fetal kidneys was obtained at 104-106 days (0.73 gestation, n = 6), and the other at 138-140 days (0.96 gestation, n = 6). Similar studies were performed in kidneys obtained from a lamb (2 weeks old) and from non-pregnant adult sheep, n = 4. Using rabbit anti-mouse renin antiserum that was proven to cross react with sheep renin and 0.033% 3',3'-diamino benzidine tetrachloride as a chromogen, immunoreactivity was found to be localized in the classical juxtaglomerular apparatus and the afferent arteriole in the immature fetuses, newborn lamb and adult sheep. In the mature fetuses a more extensive distribution was noted. Immunoreactivity was found in the afferent arteriole and the juxtaglomerular apparatus as well as other segments of the arterial vascular tree. These findings suggest that renal renin distribution in the lamb fetus is developmentally regulated. The results also correlate well with reports about renal cortical renin content and plasma renin activity at the stages studied. These observations further support the hypothesis that increased renal renin expression occurs in the fetus just prior to birth. PMID- 8713728 TI - Changes in M-CSF-like activity during chicken embryonic development. AB - We have established a method for separation of chicken bone-marrow cells using Percoll density gradient centrifugation, and have developed a new method for determining chicken M-CSF-like activity employing a liquid culture. Using this method, we determined M-CSF-like activities in egg yolk, chorioallantoic fluid (CAF) and amniotic fluid (AmF), and studied the effects of M-CSF on development of chicken embryos. M-CSF-like activity in egg yolk was at a high level before the incubation of the egg; it began to decrease on the third day of incubation and rapidly decreased on the fourth day, and no significant activity was detected after the tenth day of incubation. M-CSF-like activity in CAF was very low, and it exhibited almost no change during development. No M-CSF-like activity was detected in AmF throughout the experimental period. PMID- 8713729 TI - Effects of corticotrophin-releasing factor and vasopressin on plasma adrenocorticotrophin molecular forms, aldosterone and corticosterone in young and adult rats and rabbits. AB - Synthetic ovine corticotrophin-releasing factor (CRF) and arginine vasopressin (AVP) were injected alone or in combination (for each peptide 1 microgram/kg body weight) in 7-day-old and adult rats and rabbits. Fifteen minutes after the interscapulary injection, blood was collected for plasma adrenocorticotrophin (ACTH), corticosterone and aldosterone evaluation by RIA. The different circulating forms of ACTH were isolated by Sephadex G50 column chromatography with 1% formic acid and measured by RIA using 1-24 ACTH as standard. Such experiments were previously described in lambs and guinea-pigs using the same schedule. In young and adult rabbits the predominant circulating IR-ACTH form was 'big' ACTH; after stimulation with CRF, AVP or CRF + AVP the 'intermediate' IR ACTH was greatly increased in adults, but no change was observed in young rabbits. In young and adult rats the predominant circulating form was "intermediate' ACTH in control and injected animals; ACTH increased after CRF alone or in combination with AVP, but not after AVP alone. In both species the 'intermediate' forms of IR-ACTH were not eluted at the same time by chromatography, and calculated molecular weights were different: 14500 in rats and 9500 in rabbits. Plasma corticosterone and aldosterone were increased in rat and rabbit adults after CRF and AVP; however, they remained unchanged in young rabbits and slightly increased only after CRF in young rats in which corticosterone remained at a very low concentration compared with that in adults. Hence, the pituitary-adrenal axis of 7-day-old rabbits and rats in less reactive than that of sheep and guinea-pig of the same age. PMID- 8713730 TI - Changes in thoracic dimensions induced by breathing movements in fetal sheep. AB - The dimensions of the three major axes of the thorax (longitudinal, transverse and antero-posterior) were monitored in utero from the ultrasonic transit-time between pairs of piezo-electric transducers chronically implanted at opposite sides of the thorax in five fetal sheep at 119-122 days of gestation; tracheal and vascular catheters and diaphragmatic EMG electrodes were also implanted. To measure thoracic length, ultrasound transducers were implanted on the diaphragm and over the upper sternum and manubrium sterni. A pair was implanted on opposite sides of the chest to measure thoracic width, and another pair was implanted over the lower thoracic spine and lower sternum (antero-posterior dimension). The width of the thorax either decreased (mean 0.6 +/- 0.2 mm) or increased (mean 0.7 +/- 0.1 mm) during fetal breathing movements (FBM). The depth of the thorax (antero-posterior dimension) usually decreased (mean 0.9 +/- 0.1 mm) during FBM. The distance between the upper sternum and the diaphragm decreased by 1.0 +/- 0.1 mm (left side) and 1.6 +/- 0.3 mm (right side) during inspiratory efforts. The distance between the upper thorax and a fixed point on the lower thoracic spine decreased by a similar amount. Overall, fetal thoracic dimensions changed by 1 3%. The largest changes measured, and the most surprising, were reductions, rather than increases, in the separation between the dome of the diaphragm and the upper thorax; this suggests that, during inspiratory efforts, the upper thorax moves caudally by a greater distance than the diaphragm. FBM induce complex and variable changes in thoracic dimensions; these are likely to induce small alterations in the shape of the lungs that may act as a stimulus to lung growth. PMID- 8713731 TI - Cerebral, tympanic and colonic thermometry in the piglet. AB - The temperature of the brain is crucial for the outcome of hypoxic/ischaemic brain damage. In clinical medicine and in animal experiments involving survival after hypoxia/ischaemia, non-invasive measurement of cerebral temperature is needed. We have therefore compared tympanic and colonic temperature with cerebral temperature in the newborn piglet during hypothermia. Ten piglets aged 12-60 h were cooled to 35 degrees C (mild hypothermia) for 150 min and rewarmed. Thereafter, four of the piglets were again cooled to approximately 29 degrees C for less than one hour (moderate hypothermia). During stable mild hypothermia and normothermia the cerebro-tympanic temperature difference in individual piglets was less than +/- 0.4 degrees C (95% confidence intervals < or = 0.18 degrees C) and the cerebro-colonic temperature difference was -0.7 to 0.4 (95% confidence interval < or = 0.28 degrees C). The differences were larger during moderate than during mild hypothermia and largest during rapid changes in body temperature. Then the tympanic temperature correlated with the cerebral temperature significantly better than did the colonic temperature (95% confidence interval 0.3 to 0.3 versus -0.6 to 1.4 for the ten minutes with the least good correlation). PMID- 8713732 TI - Developmental changes in beta-adrenergic and muscarinic modulations of Ca2+ currents in fetal and neonatal ventricular cardiomyocytes of the rat. AB - Developmental changes of the beta-adrenergic and muscarinic modulations acting on the slow Ca2+ channels in rat ventricular cardiomyocytes were investigated using Ba2+ as the charge carrier (IBa(L)). Single cardiomyocytes from fetal and neonatal rats were freshly isolated, and currents were recorded by whole-cell voltage clamp in Na(+)-free, K(+)-free external solution. Basal current densities showed a large increase between fetal days 15 and 18, and a small decrease after fetal day 20. The ability of isoproterenol (ISO, 1 microM) to enhance IBa(L) increased dramatically between fetal days 18 and 20, and remained almost constant thereafter during the neonatal period. The ability of forskolin (FOR, 10 microM) to stimulate IBa(L) also increased markedly between fetal day 18 and neonatal day 1. Carbachol (CCh), in a dose-dependent manner, reversed the stimulation of the IBa(L) produced by ISO in neonatal cardiomyocytes. The IC50 values for CCh inhibition of IBa(L) was almost the same on all neonatal days tested (neonatal days 1, 5 and 10). The results indicate that there are developmental changes in beta-adrenergic modulation of IBa(L) in the late fetal period, including a large jump in sensitivity to ISO on about fetal day 19. These findings are consistent with the steep increase in number of beta-adrenergic receptors that occurs between fetal days 18 and 20 (Kojima, Sperelakis & Sada, 1990a: Kojima, Ishima, Taniguchi, Kimura, Sada & Sperelakis, 1990C). PMID- 8713733 TI - Brown adipose tissue and liver development during early postnatal life in hand reared and ewe-reared lambs. AB - This study examined the effects of modest changes in ambient temperature in hand reared lambs (experiment one) and in ewe-reared lambs (experiment two). Lambs were killed at either 8 or 31 days of age and perirenal adipose tissue was identified as being brown adipose tissue (BAT) from measurements of thermogenic activity (i.e. GDP binding to uncoupling protein in isolated mitochondria) or thermogenic capacity (i.e. detection of uncoupling protein by immunoblotting). In addition, type I and II iodothyronine 5' monodeiodonase (5'MDI) activities were assayed in perirenal adipose tissue, plus type I 5'MDI activity in liver. Plasma samples were also taken for measurements of glucose, lactate, insulin, triiodothyronine (T3) and thyroxine (T4) concentrations. In experiment one, lambs were hand-reared at either warm (WR; 25 degrees C) or cool (CR; 10-15 degrees C) ambient temperatures. Mean growth rate over the first 8 days of life in CR lambs was 88 g/day and increased to 128 g/day over the first month of life. Growth rate in WR lambs was constant at 141 g/day. Thermogenic activity of BAT was significantly higher in CR than WR lambs, but total weight and tissue lipid content of perirenal adipose tissue were significantly lower in the CR group. In both WR and CR lambs, the thermogenic activity of BAT fell by an average of 71% between 8 and 31 days. At 31 days of age, uncoupling protein in mitochondria could be detected only by immunoblotting in adipose tissue sampled from CR lambs. There was no effect of ambient temperature on type I or type II 5'MDI activity in BAT or liver; it decreased in adipose but not liver tissue between 8 and 31 days of age. The plasma concentrations of glucose, insulin and T3 tended to decline with age in CR but not in WR lambs. In ewe-reared lambs perirenal adipose tissue weight and tissue lipid content more than doubled between 8 and 31 days of age, but the level of GDP binding decreased from 85 to 5 pmol/mg mitochondrial protein over this period. Liver weight increased by 55% between 8 and 31 days of age, but hepatic 5'MDI activity remained unchanged. The plasma concentrations of T3, T4 and lactate, but not glucose or insulin, increased between 8 and 31 days of age. It is concluded that hand-rearing lambs at a cool ambient temperature significantly delays postnatal development, to the extent that BAT characteristics are retained. Ewe-rearing lambs enhances the rate at which BAT adopts the characteristics of white adipose tissue, and it prevents the postnatal decline in plasma concentrations of thyroid hormones. PMID- 8713734 TI - Effect of thyroid status in the perinatal period on oxidative capacities and mitochondrial respiration in porcine liver and skeletal muscle. AB - Regulatory thermogenesis is reduced in newborn piglets which have been made hypothyroid during late gestation by giving the sow a high glucosinolate rapeseed diet (test animals). Thereafter, the progressive increase in thermogenic capacity parallels the development of a marked postnatal hyperthyroid state. To explain these effects of thyroid hormones at the tissue and mitochondrial levels, we have examined both liver and skeletal muscle to determine possible underlying changes in (i) tissue oxidative capacities (cytochrome oxidase (CO) activity), between 80 d of gestation and 48 h after birth, and (ii) mitochondrial content and respiratory capacities at 24 h of life. In control piglets, CO activity increased sharply during late gestation and the first 2 d of life in liver and rhomboideus (RH) muscle (P < 0.01), whereas only a prenatal increase was observed in longissimus dorsi (LD) muscle. Test fetuses were hypothyroid and had lower CO activities than controls during late gestation in RH muscle (P < 0.06, at 110 d of gestation; P < 0.08, at birth) and in liver (P < 0.001, at birth). The postnatal increase in CO activity in RH muscle and liver was higher (P < 0.05) in test than in control piglets, and as a result the difference between the 2 groups was not significant by 24-48 h of life. There was no effect of treatment on LD muscle. At 24 h, hyperthyroid test piglets had lower amounts of mitochondrial proteins than controls (P < 0.05) in all three tissues, possibly reflecting reduced mitochondrial protein synthesis during fetal life and suggesting that high postnatal T3 levels did not bring about major increases in protein synthesis within 24 h. However, test piglets exhibited higher rates of mitochondrial respiration than controls in liver and RH muscle, as shown by increases in State III and FCCP-stimulated respirations (P < 0.05), and mitochondrial CO and creatine kinase activities (P < 0.05). In RH muscle, both subsarcolemmal and intermyofibrillar mitochondria showed the same trends. No changes were observed in LD muscle. Our results describe for the first time the effect of thyroid hormones on perinatal oxidative capacities and neonatal mitochondrial respiration in liver and skeletal muscle of the pig, through both the short-term regulation of mitochondrial respiration and the long-term control of mitochondrial biogenesis. The differential sensitivity of LD and RH muscles to thyroid hormones is discussed. PMID- 8713735 TI - Prediction of cerebral blood flow in fetal lambs by carotid artery ultrasonic flow transducer. AB - To determine whether common carotid artery blood flow measured with an ultrasonic flow transducer would predict brain blood flow in fetal sheep, we measured unilateral common carotid artery blood flow and compared this to simultaneous measurements of total brain blood flows made by radioisotope-labelled microsphere techniques. We studied anaesthetized, exteriorized fetal sheep with intact umbilical circulation after ligation of extracranial, extracerebral arteries and placement of a common carotid artery flow transducer; five fetuses at 120 d gestation had 19 total comparison measurements. As measured by microsphere technique, mean basal blood flow during undisturbed conditions to regional brain areas were similar to normal values reported for the exteriorized ovine fetus; these flows were highly correlated to fetal PaCO2 and successfully varied over a wide range (total brain 9.1-200.4 ml/min/100g and total cortex 6.1-153.1 ml/min/100g) in subsequent experimental conditions of hypercapnia or occluded blood flow. Blood flow as measured by flow transducer significantly correlated (P < or = 0.01) with microsphere measurements of blood flow to total brain (r = 0.56) and total cortex (r = 0.62); regional flow to cerebellum (r = 0.70) and thalamus (r = 0.60) also correlated to transducer measurements. Stronger correlations were observed at low-flow conditions to total brain (r = 0.83) and to total cortex (r = 0.90). As measured by microsphere technique, right and left cortical blood flows were highly correlated (P = 0.0001, r = 0.97), indicating that the flow transducer or surgical manipulation did not disturb the distribution of cerebral blood flow. The mean values for zero flow reference of the transducer were < 1.5% of mean basal flow values. It is concluded that the common carotid artery flow transducer technique developed in this study provides an accurate prediction of blood flow to total brain and total cortex over a wide range of values in fetal sheep. This technique provides a methodologic advantage to sequential experimental interventions and may prove advantageous to studies of fetal sheep cerebral circulation. PMID- 8713736 TI - Pulsatile release of oxytocin in newborn infants. AB - Recently, a pulsatile, basal oxytocin blood level has been described in human adults. To investigate if such a hormone profile exists already during the neonatal period, we collected blood samples continuously with 20-s fractions for 4 min in 1-day-old infants (n = 26). Plasma oxytocin concentrations were determined by a specific radioimmunoassay. The values were further analysed by the pulse detection program PULSAR. We found that 42% of the infants presented one peak in the oxytocin level during a 4-min period. The peak constituted a 111 +/- 66% (mean +/- SD) increment of the baseline value. It is suggested that the release of oxytocin during basal conditions occurs in a pulsatile way in newborn infants and that these hormone pulses reflect fluctuations in the activity of the hypothalamic neurosecretory cells. PMID- 8713737 TI - Fetal insulin-like growth factor (IGF)-I and IGF-II are regulated differently by glucose or insulin in the sheep fetus. AB - We investigated the effect of restoration of normoglycaemia or normoinsulinaemia in fetuses of starved ewes on plasma IGF-I and IGF-II concentrations. Paired maternal and fetal blood samples were taken during an initial 2-day control period, after 48 h of maternal starvation, during 24 h fetal infusion of glucose (n = 6) or insulin (n = 4) while maintaining maternal starvation and after 48 h maternal refeeding. After 48 h starvation maternal and fetal plasma IGF-I, insulin and blood glucose fell (maternal IGF-I 38.9 +/- 3.6 to 16.4 +/- 1.8 nM and fetal IGF-I 13.2 +/- 0.8 to 7.1 +/- 0.7 nM, both P < 0.05). Fetal plasma IGF II also fell (147.8 +/- 9.1 to 112.2 +/- 3.8 nM, P < 0.05), but maternal plasma IGF-II rose (71.8 +/- 6.3 to 88.8 +/- 9.2 nM, P = 0.10). Fetal glucose replacement raised fetal plasma IGF-I (11.4 +/- 1.2 nM), IGF-II (149.7 +/- 6.5 nM), insulin and blood glucose to near control values (all P < 0.05). Fetal insulin replacement raised fetal plasma IGF-I (9.0 +/- 0.6 nM) and insulin (all P < 0.05) while IGF-II (105.2 +/- 8.4 nM) and blood glucose remained depressed. Neither fetal infusion had any significant effect on maternal plasma IGF-I (13.1 +/- 1.6 nM), IGF-II (77.5 +/- 8.7 nM), insulin or blood glucose. After 48 h maternal refeeding fetal IGF-I (12.4 +/- 0.4 nM), fetal IGF-II (158.4 +/- 8.9 nM), maternal IGF-II (67.1 +/- 3.0 nM), maternal and fetal insulin and glucose had returned to near control values in both groups. Maternal IGF-I remained below control values (24.7 +/- 2.5 nM, P < 0.05). The data suggest that fetal IGF-I and IGF-II are independently regulated in the fetal circulation. While glucose plays an important role in the regulation of both IGF-I and IGF-II, the influence of glucose on fetal IGF-I is likely to be mediated by insulin, whereas for IGF-II the effect of glucose is insulin-independent. PMID- 8713738 TI - Long-term cycling of surfactant films in Wilhelmy balance. AB - Surface properties of porcine surfactant were observed during long-term cycling in Wilhelmy balance. Various amounts of surfactant were applied onto the surface as dry particles or droplets, or were suspended in the hypophase, and the films generated by this material were subjected to 50% cyclic compression at a rate of 1 cycle per min. Film spreading was faster from a droplet than from a particle of lyophilized surfactant, but the "stable period" during which minimum surface tension of the compressed film remained below 5 mN/m was significantly longer for the dry material. For surfactant suspensions the period of film "refinement", defined as the number of cycles required to reduce minimum surface tension to a level below 5 mN/m, was inversely correlated with the concentration of surfactant in the hypophase. Thirteen batches of porcine surfactant, used successfully in clinical trials for treatment of neonatal respiratory distress syndrome, were evaluated in the same system suspended in the hypophase at a concentration of 0.5 mg/ml. Films adsorbed from these batches had a median refinement period of 4 cycles, and a median stable period of 2160 cycles (36 h). In the same assay system, surfactant activity was inhibited in a dose-dependent manner by serum, fibrinogen, meconium, and bilirubin, but the specific inhibitory activity was significantly higher for bilirubin than for the other substances tested. PMID- 8713739 TI - Transvascular albumin transport and protein replenishment after haemorrhage in the chronically catheterized pregnant rabbit. AB - Pregnancy is characterized by increases in both blood and interstitial volumes, but the mechanisms are unknown. To test the hypotheses that blood volume (BV) recovery after haemorrhage in pregnant (P) is faster than in nonpregnant (NP) rabbits, and that this can be explained by a higher capillary filtration, a 20% BV haemorrhage was produced in a group of 7 P and 7 NP rabbits. We determined the BV recovery (measured by 99Tc), the total plasma protein mass and the haematocrit at 10, 20, 30, 60 and 1440 min after haemorrhage. Arterial and venous pressures and heart rate were measured during a 30-min control period, during the 10 min of haemorrhage, and for the following 70 min. In a separate group of P and NP rabbits, 125I-labelled albumin was infused and blood samples were obtained at 10, 20, and 30 min. Capillary filtration was expressed as the percentage of radioactive albumin that left the intravascular space after 30 min. No differences were found in the rate of BV recovery, arterial and venous pressures, or heart rate response between P and NP animals (P > 0.1). By 24 h total plasma protein mass (TPPM) was significantly increased above baseline in the P rabbits by an average+SEM of 13 + 3.7%, but not in NP rabbits (5 + 4.5%). At 30 min, the amount of labelled albumin in the intravascular space decreased by 15.4 + 3.4% for Pv. 8.2 + 1.2% for NP rabbits (P < 0.05). There was a high correlation between TPPM recovery and BV recovery in the P (r = 0.96) and NP rabbits (r = 0.95). In conclusion, we did not find differences in the rate at which the BV recovered during pregnancy, despite the increased capillary permeability. During pregnancy, the combination of the latter with an increase in the TPPM elevation above pre-haemorrhage levels suggests a faster rate of protein mobilization. PMID- 8713740 TI - Developmental characteristics in the daily rhythm of norepinephrine concentration within rabbit brainstem regions. AB - To examine the development of daily variations in norepinephrine levels, norepinephrine concentrations were measured within five distinct brainstem regions in 3-day-old, 21-day-old, and adult rabbits at 6-h intervals throughout the day. Norepinephrine was measured by radioenzymatic assay, and norepinephrine concentration was expressed relative to wet tissue weight. The data suggest that daily variations for norepinephrine concentrations are established by the third day of life. In the brainstem as a whole, there was an early nocturnal peak (2130 hours) for 3-day-old animals in contrast to a late nocturnal peak (0330 hours) for 21-day-old animals. Adult animals showed a late diurnal (1530 hours) peak. These gross daily variations constitute the sum of distinct region-specific patterns in the development of daily variations in norepinephrine concentration. Norepinephrine is involved in cardiorespiratory regulation and in the regulation of sleep/wake cycles. The observed developmental patterns may relate to the maturation and integration of these physiologic processes. PMID- 8713741 TI - ACTH-like bioactivity and immunoactivity in fetal lamb pituitaries at 0.65 and 0.95 gestation. AB - We wished to determine if the concentration of bioactive ACTH-like activity increased during development and if there was heterogeneity in ovine fetal anterior pituitary ACTH activity as measured by bioassay and radioimmunoassay (RIA). We obtained anterior pituitaries from eight sheep fetuses (four at 0.65 and four at 0.95 gestation; term 145 +/- 5 days) and extracted and homogenized them in ice-cold 5N acetic acid, 0.3% phenylmethanesulfonyl fluoride (PMSF) and 0.2% BSA. Fractionation of each pituitary extract was performed by size-exclusion chromatography using Sepadex G-50. The ACTH-like immunoactivity (ALI) profile for each pituitary showed two well-defined peaks. One eluted with human ACTH1-39 and the other eluted with the high molecular weight fraction in the void volume. Four fractions from the first peak representing the high molecular weight forms of ACTH activity and four fractions from the second peak representing the low molecular weight forms of ACTH activity were pooled separately. These two pools were subjected to reverse-phase chromatography (RPC) on a C-8 column using a linear gradient of 70% acetonitrile in 0.8% trifluoroacetic acid over a 60 min period. Based upon the RIA, the high molecular weight forms of ACTH from the G-50 column were resolved into three main fractions, one eluting similar to the standard ACTH1-39 and the remaining two eluting after that. The low molecular weight forms of ACTH from the G-50 column were resolved into three peaks, before, with, and after the standard. We used collagenase-dispersed rat adrenal cells to test the ACTH-like bioactivity (ALB) of the crude extracts and of the different fractions obtained from the RPC of the high and low molecular weight material. The concentration of ACTH-like bioactivity in the crude extracts was similar at the two stages of gestation. However, there was a trend for the low molecular weight peak to have more peptide eluting with human ACTH1-39 and higher ratios of ALB/ALI than did the high molecular weight peak. These results suggest that multiple ACTH molecular forms with different ALB/ALI ratios are present in the ovine fetal pituitary and that there is no selective increase in ACTH1-39 concentration in the fetal pituitary in late gestation. PMID- 8713742 TI - IRF-1 and IRF-2 as regulators of the interferon system and cell growth. AB - Interferons (IFNs) are families of cytokines which have been discovered and extensively characterized in the context of host defense against viral infections. We have discovered two structurally related transcription factors, Interferon regulatory factor-1 (IRF-1) and IRF-2. These two factors, however, function not only as regulators of the IFN system, but are also key transcription factors in the regulation of cell cycle and apoptosis. These studies uncover a complex gene transcription network, by which the fate of cellular responses are determined depending on how the IRF transcription factors function in conjunction with other factors, and on the promoters of distinct genes under different conditions of the cells. PMID- 8713743 TI - A technique for identification of cellular proteins by western immunoblotting and immunoautoradiography of 2-D PAGE. AB - A major effort of our work has been devoted to the identification, using two dimensional polyacrylamide gel electrophoresis (2-D PAGE), of lymphoid polypeptides that are involved in lymphoid proliferation or differentiation. We have encountered problems during this effort pertaining to the exact localization of a polypeptide(s) in the silver stained gels that is recognized by western immunoblotting or proteins detected by autoradiography. In this paper, we present a method, using India ink stained/immuno-staining replica of 2-D gel nitrocellulose membrane (NCM) or India ink stained coupled with autoradiography in the case of phosphoproteins, which allows us to exactly localize the polypeptide spots detected by these methods in the silver stained 2-D PAGE. This method is expected to popularize and widen the use of 2-D PAGE technology in the investigation of cellular polypeptides. PMID- 8713744 TI - Efficacy of passive transfer of proteins from preparative polyacrylamide gels to nitrocellulose membranes. AB - Passive transfer of proteins from preparative polyacrylamide gels to nitrocellulose membrane (NTC) was investigated. The transfer was carried out without the removal of sodium dodecyl sulphate from the denaturing gels. The transfer was quick, easy to perform, and does not require any apparatus. More than one replica of the same band can be obtained by successive transfer to more than one NTC without significantly affecting the Coomassie Brilliant Blue (CBB) staining ability of protein bands left in the gel. A fraction of protein transferred passively in the absence of electrical current from as low as 1.5 micrograms of protein per band in a 2 mm thick gel was found sufficient for detection with amido black staining, and more so by immunostaining. The method was found highly satisfactory for seven proteins (14 kDa to 66 kDa) checked in the present study and can be adopted, after checking its efficacy for any protein, for routine qualitative tests. PMID- 8713745 TI - A direct ELISA technique to detect antibodies against polioviruses. AB - A direct ELISA test was developed to detect circulating antibodies specific to poliovirus types 1, 2 and 3. Specificity of the test was established by using known polio negative and positive sera. The assay was compared with the standard microneutralization test using sera from polio and non polio cases vaccinated and unvaccinated subjects and standard positive sera obtained from WHO/EPI (Geneva). The reproducibility of the results was tested using different batches of the antigen prepared from Sabin strain of poliovirus supplied by ERC Bombay and from Sabin strain of virus obtained from WHO/EPI. ELISA was found to be as sensitive as microneutralization test in detecting seronegatives and was found to be specific to polio by giving negative results with non polio cases. ELISA is thus a rapid and simple method that may be useful for mapping seroepidemiology of poliomyelitis and as a tool for effective surveillance of the disease by offering rapid diagnosis in acute cases. PMID- 8713746 TI - A spin decay assay for tumor necrosis factor cytotoxicity. AB - Tumor necrosis factor (TNF) is believed to elicit its response primarily by an oxidative process. We present a sensitive biophysical assay system to monitor TNF induced cell killing by electron paramagnetic resonance (EPR) spectroscopy using the spin label 2,2,6,6-tetramethyl-1-piperidine-n-oxyl (TEMPO). The rate of TEMPO spin decay by the TNF sensitive L929 cells was hypothesized to be an indicator of TNF cytolytic response. The cell-induced rate of loss of TEMPO signal was sensitive to concentration as well as time of incubation with TNF. The TEMPO spin decay first derivative plot correlated well with corresponding TNF cytotoxicity curve as obtained by a standard tetrazolium dye reduction bioassay. While dimethyl sulfoxide (DMSO) (up to 500 mM) and alpha-tocopherol (1 mM) inhibited the TNF induced inhibition of rate of loss of TEMPO decay, superoxide dismutase (SOD) (100 U/ml), catalase (1000 U/ml), and histidine (6 mM) had little effect on the TEMPO decay rate. The loss of TEMPO signal was determined to be at an outer domain of the lipid bilayer as illustrated by its decay and broadening in presence of NiCl2. Thus the reduction of a lipid soluble spin label like TEMPO as monitored by EPR could be a useful reporter to study TNF cytotoxicity in target cells. PMID- 8713747 TI - Activity of alkaline phosphatase in water-in-oil microemulsions containing vegetable oil. AB - The hydrolysis of p-nitrophenyl phosphate by the enzyme alkaline phosphatase has been studied in vegetable oil containing water-in-oil (W/O) microemulsions of six different compositions at four different (water)/(surfactant) mole ratios of 10, 17.6, 24.7 and 37. The vegetable oils used are ricebran oil (RO) and clove oil (CO) and the amphiphiles used are Aerosol OT (AOT), cinnamic alcohol (CA) and Tween-20 (T-20). The hydrolytic process does not follow conventional Michaelis Menten equation normally observed for enzymatic process. In the water/vegetable oil microemulsions, the enzyme seems to lose its activity when AOT is the amphiphile. The amount of p-nitrophenol generated as a result of hydrolysis is independent of the presence of the enzyme. With Tween-20 as the amphiphile, the microemulsion produces an initial retarding effect which ultimately gets appreciably compensated. PMID- 8713748 TI - Molecular inactivation of testicular hyaluronidase in solid state after proton irradiation: a study based on target size, substrate binding and thermodynamic analysis of heat denaturation. AB - Dose response activity curve of testicular hyaluronidase (HDase) following proton irradiation in dry state follows complicated mechanisms which may involve multiple hits and multiple targets of variable sizes giving a constant G value of 1.66. Target analysis appears to be modified by slow recovery of activity when irradiated enzyme is brought to aqueous phase. However, pattern of irradiation at a dose of 1 x 10(5) to 8 x 10(5) Gy reveals that though binding affinity of enzyme to the substrate (hyaluronic acid) increases as shown by declining Km from 500 mg/l to 300-70 mg/l, the reaction rate of catalysis by irradiated HDase is decreased due to decrease in reaction velocity (Vmax: 266 versus 76 units at 8 x 10(5) Gy). Activation analysis of heat denaturation of nonirradiated HDase suggested the involvement of 78 kcal/mole of energy of activation (E*a) which declined to 63-52 k cal/mole after irradiation at 1 x 10(5) to 8 x 10(5) Gy for residual enzyme. The corresponding change in entropy of activation (delta S*) increased from a control value of -291 eu to -236 eu at 8 x 10(5) Gy. From thermodynamic analysis in association with recovery in aqueous phase, it is concluded that HDase is inactivated due to dissipation of proton energy among weak forces including H bonds associated with secondary/tertiary structure of molecules. PMID- 8713749 TI - Effect of Ca(2+)-channel blockers, Ca(2+)-ionophore and increased pyrene excimer formation on the microsomal glucose-6-phosphatase. AB - The dependence of microsomal glucose-6-phosphatase (G-6-Pase) activity on Ca2+ as well as the membrane lipid microviscosity was studied by the effect of Ca(2+) channel blockers (namely verapamil and nifedipine), Ca(2+)-ionophore, A23187 and pyrene excimer formation. Channel blockers depressed the G-6-Pase and Ca(2+) ATPase while the ionophore increased these activities. Dimethyl sulfoxide, a known membrane surface active agent showed no change. Ca(2+)-uptake into the membrane has expectedly been lowered by the channel blockers while the ionophores facilitated the ion flux. Excimer formation of the fluorescent probe, pyrene as an indicator of increased membrane fluidity, and microviscosity calculated from there on, showed that Ca(2+)- and lipid microenvironment in the membrane significantly influenced the activity of G-6-Pase. Membrane lipid composition such as phospholipid/cholesterol molar ratio which also indicates an increased membrane fluidity is markedly increased with the ionophore but decreased with the channel blockers, while protein/phospholipid ratio remained unchanged. Microsomal G-6-Pase is a multicomponent multifunctional protein. It is argued that Ca2+ may play the role of an obligatory cofactor not only for the hydrolysis of G-6-P (catalytic part of the enzyme) but also involved in the regulation of substrate and product transport in or out of the endoplasmic reticulum lumen. PMID- 8713750 TI - Physico-chemical properties of brain cathepsin H. AB - Cathepsin H (EC 3.4.22.16) from cow brain, purified to approximately 1800-fold with approximately 26% activity yield, hydrolysed BANA, Leu-2-NNap, Arg-2-NNap, and Met-2-NNap maximally at pH 6.5, 6.8, 7.0 and 7.2, respectively. It was activated by sulphydryl compounds and EDTA while sulphydryl alkylators and blockers were found to inhibit the enzyme activity. Met-2-NNap was found to be the best substrate followed by Thr-2-NNap, His-2-NNap, Leu-2-NNap, Arg-2-NNap and Ala-2-NNap, respectively. The Km values for hydrolysis of various substrates viz., Met-2-NNap, Leu-2-NNap, Arg-2-NNap, Arg-NNapOMe, Thr-2-NNap, His-2-NNap, BANA, Arg-pNA and Lys-pNA were 0.128, 0.167, 0.169, 0.288, 0.428, 0.500, 0.667, 0.195 and 0.476 mM, respectively. The temperature optima for hydrolysis of BANA and Leu-2-NNap were approximately 45 degrees C and approximately 50 degrees C with activation energies of approximately 13.7 and approximately 11.0 kcal mole 1, respectively. The enzyme was fairly stable upto 50 degrees C and between pH 4.0-7.5. PMID- 8713751 TI - Incorporation of synthetic tetramethylene diamine 8,8'-bis(riboflavin tetraacetate) in DMPC vesicles. AB - The tetramethylene diamine 8,8'-bis(riboflavin tetraacetate) is synthesized by condensation of 8 alpha-bromo riboflavin tetraacetate and 1,4 diaminobutane. The incorporation of bis(riboflavin tetraacetate) in bilayer of DMPC vesicles has been studied by gel permeation chromatography, UV-visible and fluorescence spectroscopy. PMID- 8713752 TI - Lipid composition and membrane fluidity of monkey small intestinal brush border membrane: regional differences. AB - Brush border membrane vesicles (BBMV) isolated from various regions of monkey small intestine were compared for lipid composition, membrane fluidity and sodium dependent D-glucose transport. Total phospholipid content was same in all regions whereas cholesterol content was high in duodenum and jejunum as compared to ileum resulting in higher cholesterol/phospholipid molar ratios. Regional differences in individual phospholipid subclasses and fatty acids in total lipids were also observed. Fluidity measurements showed that the membranes of duodenum and jejunum were less fluid than ileum. The change in BBMV fluidity appears to be due to alteration in cholesterol/phospholipid ratio and phospholipid composition. The sodium dependent D-glucose uptake was more in duodenum and jejunum as compared to ileum. These results suggest that there is a regional difference in the lipid composition and fluidity of BBMV in monkey small intestine which may influence their function. PMID- 8713753 TI - Synthesis and phosphorylation of macrophage membrane proteins in protein deficient rat. AB - Changes in the biosynthesis and phosphorylation of rat peritoneal macrophage membrane proteins induced by protein malnutrition have been studied. The results clearly indicate that the biosynthesis of high molecular weight proteins (45-200 kDa) and their phosphorylation are significantly reduced in the macrophages isolated from protein deficient (4% protein-fed) rats compared to the control group fed 20% protein diet. Lipopolysaccharide (LPS) treatment both in vivo and in vitro enhanced the synthesis and phosphorylation of these proteins in both control and protein deficient groups; however, the extent of enhancement was much less in the deficient group. These results indicate that besides the down regulation of these membrane proteins, protein malnutrition seems to make these macrophages less responsive to potent immuno stimulants like LPS. PMID- 8713754 TI - Correlation of biological activity (therapeutic and toxic) with solvochromic properties of metronidazole, emetine hydrochloride and diloxanide furoate. AB - Goat blood, when incubated for different periods with diloxanide furoate, metronidazole and emetine hydrochloride, underwent changes in fatty acid constituents and their peroxidation products measured as malonaldehyde. These findings, together with the changes noted in the drug-lipid partition coefficient, are discussed in an attempt to correlate the lipid constitution and biological activity of the drugs. PMID- 8713755 TI - Withaperuvin-E, a new free radical scavenger isolated from Physalis peruviana. PMID- 8713756 TI - The road to professionalism in medical informatics: a proposal for debate. PMID- 8713757 TI - Potholes in the road to professionalism in medical informatics. PMID- 8713758 TI - The synergism of health/medical informatics revisited. PMID- 8713759 TI - Theory and practice of medical informatics professionally balanced in problems and projects. PMID- 8713760 TI - "The road to professionalism in medical informatics: a proposal for debate". PMID- 8713761 TI - Medical informatics: a key to the future of medicine and health care? PMID- 8713762 TI - Managing different time granularities of clinical information by an interval based temporal data model. AB - In the field of databases, time management at different levels of granularity has been an issue for several years, for instance when dealing with clinical information from different databases using different time units, dealing with natural language expressions, or when dealing with temporal uncertainty. A temporal data model is proposed to manage the temporal aspect of data, presented at various and mixed levels of granularity. The concept of temporal assertions shapes the entire temporal information. The model provides a temporal dimension to the data by using intervals that can be specified at different granularities. The model supports a three-valued logic, where True, False and Undefined are the truth values. The temporal data model allows to manage some degrees of uncertainty when establishing temporal relationships between intervals or between temporal assertions, expressed at different granularities. The logical connectives and quantifiers can manage each of the three truth-values. We applied the temporal data model by implementing an object-oriented database system for managing follow-up clinical data from patients who underwent percutaneous transluminal coronary angioplasty. PMID- 8713763 TI - A real time control architecture for continuously managing patients in a care unit. AB - The monitoring and treatment of patients in a care unit is a complex task in which even the most experienced clinicians can make errors. A hemato-oncology department in which patients undergo chemotherapy asked for a computerized system able to provide intelligent and continuous support in this task. One issue in building such a system is the definition of a control architecture able to manage, in real time, a treatment plan containing prescriptions and protocols in which temporal constraints are expressed in various ways, that is, which supervises the treatment, including controlling the timely execution of prescriptions and suggesting modifications to the plan according to the patient's evolving condition. The system to solve these issues, called SEPIA, has to manage the dynamic, processes involved in patient care. Its role is to generate, in real time, commands for the patient's care (execution of tests, administration of drugs) from a plan, and to monitor the patient's state so that it may propose actions updating the plan. The necessity of an explicit time representation is shown. We propose using a linear time structure towards the past, with precise and absolute dates, open towards the future, and with imprecise and relative dates. Temporal relative scales are introduced to facilitate knowledge representation and access. PMID- 8713764 TI - Digital optical archiving of medical records in hospital information systems--a practical approach towards the computer-based patient record? AB - The large number of inpatients and outpatients in university hospitals leads to high costs of medical documentation and to an increasing number of medical documents. Due to legal regulations, these medical records have to be stored for 30 years. This implies spatial, organizational, and economical problems. At present, conventional archiving in hospitals often does not satisfy the need to make medical records available for health-care professionals in a systematic and timely manner. From 1989 to 1993 a pilot study on "digital optical archiving of medical records" was carried out at Heidelberg University Hospital. The study has shown the feasibility of digital optical archiving in hospital s if done under certain conditions. In 1995, Heidelberg University Hospital adopted a procedure for "digital optical archiving of medical records". The digital optical archive will first be filled with the medical records of the department of neurosurgery and the endoscopic and echographic images and reports of the department of internal medicine. It is to be expected that this procedure will gradually lead to an integrated functionality on health-care professional workstations, to a hospital-wide use of an electronic patient record, and to media-independent document management systems. The paper focuses on the potentials of digital optical archiving as an integral part of hospital information systems, and on the requirements for the systematic managements of hospital information systems with respect to digital optical archives. PMID- 8713765 TI - Utilization of neural network in the elaboration of an evaluation scale for pain in cerebral palsy. AB - An interesting aspect of neural networks is shown in the elaboration of an evaluation scale for pain in cerebral palsy with severe mental retardation. Because of the diversity of cases, the number of items had to be limited in the final step of statistical validation. Classical analysis on prior data did not allow to decide whether the variability in results is more likely due to the type of disability (i.e., the possibility of pain expression) than to the actual presence of pain. A neural network was used to find implicit relations between the data, with the advantage of having total control on the variables' status by applying variations in the network architecture. This allowed for the rapid identification more significant item combinations as a function of degree of relationship to pain in cerebral palsy. PMID- 8713766 TI - Correction of risk estimates for measurement error in epidemiology. AB - Epidemiologists are under considerable pressure to acknowledge the presence of measurement error in the determination of risk factors. Repeatability and validation studies are often prescribed in conjunction with epidemiological studies. We describe some practical uses for repeatability and validation study data, in terms of correcting risk estimates for measurement error. Commonly available methods are described, with their advantages and shortcomings. A user friendly computer program to carry out the analyses described accompanies the paper. PMID- 8713767 TI - Methodology for the construction of a disease nomenclature and classification system for clinical use. AB - The nature and problems of the linguistic representation of clinical data are discussed, using the linguistic theory of Ferdinand de Saussure. Based on the conclusions, a methodology for the construction of a disease nomenclature and a classification system, suitable for use in clinical information systems, is developed using set theory. PMID- 8713768 TI - The CARE telematics network for the surveillance of influenza in Europe. AB - Since the 1950s, national networks for the surveillance of influenza have been progressively implemented in several countries. New epidemiological arguments have triggered changes in order to increase the sensitivity of existent early warning systems and to strengthen the communications between European networks. The WHO project CARE Telematics, which collects clinical and virological data of nine national networks and sends useful information to public health administrations, is presented. From the results of the 1993-94 season, the benefits of the system are discussed. Though other telematics networks in this field already exist, it is the first time that virological data, absolutely essential for characterizing the type of an outbreak, are timely available by other countries. This argument will be decisive in case of occurrence of a new strain of virus (shift), such as the Spanish flu in 1918. Priorities are now to include other existing European surveillance networks. PMID- 8713769 TI - Age-related reference ranges for growth parameters. AB - We propose an approach to the computation of age-related reference ranges which guarantees that the proportion of sampled data points covered by the reference band has an arbitrarily specified value. Unlike other approaches in the literature, this method does not have to rely on parametric distributional assumptions. The boundaries of the bands are given by smooth curves, and for modeling the mean as a function of age, a sufficiently flexible class of monotonically increasing functions is used, since in the applications all measured variables are biological growth parameters. The width of the reference band is allowed to be proportional to any suitably fixed function of age which we propose in practice to be taken linear. The approach is illustrated by examples from a large-scale study of the distribution of fetal size measurements obtained by routine prenatal sonography. PMID- 8713770 TI - Aprotinin in aortocoronary bypass surgery: increased risk of vein-graft occlusion and myocardial infarction? Supportive evidence from a retrospective study. AB - To assess the thrombotic risk of aprotinin in aortocoronary bypass surgery, we retrospectively analyzed the results of a trial, originally designed to compare the effects of one-year treatment with various antithrombotic drugs in the prevention of vein-graft occlusion. Graft patency at one year was assessed by angiography. Myocardial infarction, thromboembolism, major bleeding, and death were clinical endpoints. Of 948 randomized patients, 42 received aprotinin, all enrolled by one of the participating centres. Occlusion rates of distal anastomoses were 20.5% in the aprotinin group and 12.7% in the non-aprotinin group (p = 0.091). The proportions of patients with occluded grafts were 44.1% versus 26.3% (p = 0.029). Perioperative myocardial infarction occurred in 14.3% and 7.0%, respectively (p = 0.12). Mean postoperative blood loss was 451 ml in the aprotinin group compared with 1039 ml in the non-aprotinin group (p < 0.0001). Mean transfusion requirements were 1.1 U versus 2.1 U of red blood cells (p = 0.004). Aprotinin decreases blood loss and transfusion requirement. Our data suggest that this benefit may be associated with a reduction of graft patency and an increased risk of myocardial infarction. PMID- 8713771 TI - The 1691 G-->A mutation in the factor V gene: relationship to activated protein C (APC) resistance and thrombosis in 65 patients. AB - Four hundred fifty subjects were screened for the 1691 G-->A mutation in the factor V gene. Two hundred ninety-seven patients were referred to us for unexplained thrombosis, 133 were family members of these patients and 20 were normal subjects. We studied the relationships between the mutation, resistance to APC and thrombosis. Among the 450 subjects tested, 65 belonging to 42 families were found to have the 1691 G-->A mutation in one (n = 61) or both alleles (n = 4). The prevalence of the mutation in the thrombotic patients was 13%. Resistance to APC was tested for in 247 subjects not on anticoagulant treatment (4 homozygous and 44 heterozygous for the mutation, and 199 individuals without the mutation). Incomplete cosegregation of heterozygosity for the 1691 G-->A mutation with APC resistance (APC-SR < 2.4 or n-APC-SR < 0.75) was observed, showing that the functional assay alone is insufficient for a firm diagnosis. In patients carrying the mutation, elevated levels of prothrombin fragment 1 + 2 and D-dimers pointed to increased thrombin generation in vivo. Clinical manifestations in the heterozygous subjects were very similar to those reported in heterozygous PC or PS deficiencies, but the first thrombotic event occurred later than in PC- or PS deficient patients. Homozygosity for the factor V gene mutation appears to be a far more benign thrombotic disorder than homozygous PC and PS deficiencies. PMID- 8713773 TI - The effect of cigarette-smoking on cardiovascular risk factors: a study of monozygotic twins discordant for smoking. AB - The association of cigarette smoking with the development of occlusive vascular disease is firmly established. Unfavourable changes in a series of variables held independent risk factors for the development of vascular lesions (HDL cholesterol, haematocrit, white blood cell count, fibrinogen and plasminogen activator inhibitor-1 (PAI-1)) are thought to be directly influenced by cigarette smoking. However, the role played by the genotype in the effect of smoking on the above parameters has not been investigated. To control the genotype, we studied the relationship between cigarette smoking and a series of cardiovascular risk factors in 27 monozygotic twin pairs (7 male and 20 female pairs, mean age +/- SD: 47.4 +/- 12.9 yrs) with a life-long discordance for smoking. Smoking twins had a life-long dose of smoking (Brickman index) of 287.3 +/- 241.5. Body mass index, blood pressure, haematocrit, haemoglobin and red blood cell counts, total cholesterol levels and the acute phase reactants alpha 1-acid glycoprotein and C reactive protein were similar in smokers and non-smokers. Triglyceride was higher by 12.6% (9.5-35%, 95% confidence interval, p = 0.02) and HDL-cholesterol lower by 7.5% (0.2-15%, p = 0.04) in the smoking co-twins, who also had 8.4% (-0.2-17%, p = 0.06) higher white blood cell counts and 4.1% (1.2-7%, p < 0.01) larger mean platelet volume. There was no significant difference in clottable fibrinogen (by two methods) or in the activity of plasminogen activator inhibitor-1 between the two groups, nor was the within-pair difference in these parameters related to the smoking dose. Echo-doppler examination of the carotid arteries of 24 twin pairs showed mostly minor atherosclerotic lesions in 46% and 42% of the smoking and non smoking co-twins. After adjustment for age, systolic blood pressure and platelet count and volume were the only variables significantly associated to the presence of vascular lesions. Cigarette smoking is associated with an atherogenic lipid profile and with changes in platelets and white cells potentially reflecting endothelial cell damage. When controlling the genotype, fibrinogen and PAI-1 activity levels did not seem directly influenced by cigarette smoking. PMID- 8713772 TI - Contribution of a new, rapid, individual and quantitative automated D-dimer ELISA to exclude pulmonary embolism. AB - The performance of a new automated ELISA for a rapid, individual and quantitative measurement of plasma D-dimer (VIDAS D-dimer) has been evaluated. First, a study of 100 patients was performed in order to choose the best couple of antibodies in comparison with an already clinically validated ELISA. Then the results were certified in a prospective study including 195 consecutive patients suspected of pulmonary embolism (PE). For a cut-off level of 500 ng/ml VIDAS D-dimer showed a sensitivity of 100% (95% confidence interval 92-100), a specificity of 37.6%, a negative predictive value of 100% (95% CI 93.3-100) and a positive predictive value of 33.1%. During a 6 months' follow-up no patient (95% CI 0-6.4) with D dimer < 500 ng/ml presented a new suspicion of venous thromboembolic disease. These results suggest that this rapid and single-dose ELISA provides a very useful tool for the clinician to exclude on a day-to-day basis the diagnosis of PE. PMID- 8713774 TI - Smoking, haemostatic factors and the severity of aorto-iliac and femoro-popliteal disease. AB - To determine relationships between haemostatic and rheological factors and severity of peripheral atherosclerosis and differences by site, an angiographic cross-sectional survey was carried out on 192 men and women with intermittent claudication or rest pain. 34 patients were classified as having aorto-iliac disease, 85 femoro-popliteal disease and 73 dual-site disease. Mean levels of haemostatic or rheological factors did not differ significantly between the three site groups. In all 192 patients, disease severity in the femoro-popliteal segments was correlated with plasma nephelometric fibrinogen (r = 0.20, p < or = 0.01), von Willebrand factor (r = 0.14, p < or = 0.05) and fibrin D-dimer (r = 0.22, p < or = 0.001). On multiple regression analyses, fibrinogen was independently associated with disease severity in the femoro-popliteal segments (p < or = 0.05), but not in the aorto-iliac segments. Adjustment for packyears or serum thiocyanate had little effect on the association of fibrinogen with severity of disease. An inverse relationship between plasminogen activator inhibitor and disease severity in the femoro-popliteal segments was found only in men (r = 0.24, p < or = 0.01). We conclude that elevated fibrinogen and disturbed fibrinolytic activity may be related to the extent of disease within the femoro popliteal arteries, more so than in the aorto-iliac arteries. PMID- 8713775 TI - Safety profile of porcine factor VIII and its use as hospital and home-therapy for patients with haemophilia-A and inhibitors: the results of an international survey. AB - A multicentre retrospective survey was conducted to re-assess the use of porcine factor VIII (HYATE:C), its side effects and the selection of patients for regular or home-therapy. 15,152,000 units of HYATE:C were used by 154 patients. The median inhibitor cross-reactivity to porcine VIIIC of 137 patients was 15%, 27% of patients lacking cross-reactivity. An absent, intermediate or brisk specific antiporcine anamnestic response was observed in 29, 40 and 31% of patients respectively. Seven patients were treated on-demand as home-therapy for a median 6.2, range 1.5-13 years, 23 further patients were treated regularly in hospital for a median of 3, range 2-7 years. This group used 8,319,000 U of porcine VIIIC for 2,000 bleeding episodes. The incidence of transfusion reactions was 0.001%, 0.64% and 2.3%, for domiciliary infusions, infusions in multiply treated in patients, and unselected in-patient infusions, respectively. The risk of reactions was dose-related. A post-infusion fall in platelet count was common, but usually transient and clinically insignificant. This was also dose-related (r = -0.64, p = 0.002). Marked reductions in platelet count were occasionally seen, usually with intensive replacement therapy. The relative lack of side effects observed amongst patients treated at home is attributable to the low, median 33 U/kg, dose used by this group. A subgroup of inhibitor patients, identifiable by their absent or modest anamnestic response to porcine factor VIII may be treated regularly and safely with this product in small doses, over a period of years. PMID- 8713776 TI - Safety and efficacy of monoclonal antibody purified factor IX concentrate in previously untreated patients with hemophilia B. AB - The safety and efficacy of a monoclonal antibody purified factor IX concentrate were evaluated in two continuing trials of 32 previously untreated patients with mild, moderate, or severe hemophilia B. Patients were evaluated every 2 weeks for 24 weeks and every 3 months thereafter for at least 1 year. No patients became positive for human immunodeficiency virus antibody or hepatitis C virus antibody during the trial. Two patients developed a false-positive hepatitis B core antibody, one transiently, but neither had elevated levels of alanine aminotransferase (ALT). None of the 25 patients evaluable for non-A, non-B, non-C hepatitis by strict International Society of Thrombosis and Hemostasis criteria developed elevated levels of ALT indicative of posttransfusion infection. Anaphylaxis occurred in one subject who also developed an inhibitor to factor IX (19.3 Bethesda units). Five of the eight adverse events reported (63%) were mild in severity, and the relationship of three of these to therapy was considered remote. Hemostasis with monoclonal antibody purified factor IX concentrate was excellent in all patients. PMID- 8713777 TI - Effect of heterologous factor V heavy chain sequences on the secretion of recombinant human factor VIII. AB - Factor VIII and factor V share a repetitive domain structure of A1-A2-B-A3-C1-C2. To define the region(s) within the factor VIII heavy chain that result in inefficient expression of the recombinant protein, we expressed a series of factor VIII/factor V chimeras that contained heterologous sequences from the A1 and/or A2 domains. Substitution of the factor VIII A1 domain dramatically reduced secretion of factor V approximately 500-fold, whereas substitution of the factor VIII A2 domain had minimal effect on secretion. Conversely, substitution of the factor V A1 domain increased secretion of factor VIII approximately 3-fold, whereas substitution of the factor V A2 domain actually reduced secretion approximately 4-fold. Pulse chase experiments confirmed that reduced expression levels were due to decreased secretion rather than instability of secreted protein. Smaller substitutions did not further localize within the A1 domain the regions responsible for inefficient secretion. PMID- 8713778 TI - Detection of new polymorphic markers in the factor V gene: association with factor V levels in plasma. AB - Three novel polymorphisms were found in the repeated region of the large exon 13 of factor V gene, one giving rise to a codon dimorphism (Ser1240) and two causing aminoacid substitutions (His1299Arg, Leu1257Ile). An increasing frequency of the Arg1299 (R2 allele) correlated with a decreasing mean plasma factor V activity in the groups of subjects under study, which included 26 unrelated subjects with partial factor V deficiency. Family studies supported the co-inheritance both of low factor V activity and of R2 allele. The reduction of factor V activity associated with the R2 allele was not clinically symptomatic even in the homozygous condition and was characterized by a parallel reduction of antigen in plasma, in which abnormal molecules were not detected. Data suggest that the R2 allele represents a marker in linkage with an unknown defect rather than a functional polymorphism. These studies provide the first evidence of a genetic component in determining factor V levels in plasma and of a genetic linkage between the factor V gene and factor V deficiency. They also define specific haplotypes which are associated with factor V deficiency or with APC resistance (Arg506Gln) and are valuable tools for the study of factor V defects. PMID- 8713779 TI - beta 2-Glycoprotein I modulates the anticoagulant activity of activated protein C on the phospholipid surface. AB - The objective of this study was to determine whether beta 2-glycoprotein I (beta 2GPI) has procoagulant activity by inhibiting the anticoagulant activity of activated protein C (APC). beta 2GPI inhibited significantly the APC-catalyzed inactivation of factor Va in an assay using factor V-deficient plasma and physiological levels of protein S and factor Va. This inhibitory effect was diminished by the addition of increasing concentrations of phospholipids, suggesting that beta 2GPI competitively inhibits the binding of APC to the phospholipid surface. beta 2GPI inhibited weakly factor Va- and phospholipid dependent prothrombinase activity at concentrations similar to those to inhibit APC activity. The depletion of beta 2GPI from plasma led to only a slight shortening of the diluted Russell's viper venom-dependent clotting time, but to a strong and significant potentiation of the anticoagulant activity of APC. These results suggest that under certain physiological conditions beta 2GPI has procoagulant property by inhibiting the phospholipid-dependent APC anticoagulant activity. PMID- 8713780 TI - Quantification of circulating activated protein C in human plasma by immunoassays -enzyme levels are proportional to total protein C levels. AB - We have developed a simple assay that measures the circulating activated protein C (APC) in plasma. The assay requires collection of duplicate blood samples, one in citrate plus heparin and the other in citrate plus inhibitors of the enzyme. In the heparin tube, APC reacts completely and irreversibly with its major plasma inhibitors, protein C inhibitor (PCI) and alpha 1-antitrypsin (alpha 1AT), and the complexes formed are measured by ELISAs. The amount of circulating APC is calculated from the difference between the total amount of complexed APC (sample in citrate plus heparin) and the amount of APC complexed in vivo (sample in citrate plus inhibitor). Over 95% of the APC added to blood collected with heparin was recovered in the assay. The assay can easily be performed in four hours, and had a detection limit of 0.1 ng/ml APC. The mean APC level in 18 protein C heterozygous members from seven kindreds was significantly lower (0.6 +/- 0.3 ng/ml) than in 20 healthy controls (1.1 +/- 0.3 ng/ml) (p < 0.001), whereas the mean level in 10 non-affected members from the kindreds studied was 1.5 +/- 0.3 ng/ml. In the group of 12 nonanticoagulated heterozygous protein C deficient individuals, the three patients with a history of venous thrombosis had a mean APC level significantly lower than the nine asymptomatic members (p < 0.01), both subgroups showing similar protein C levels. There was a significant correlation in all groups between the levels of APC and the levels of protein C antigen (r = 0.758, p < 0.0001) and activity (r = 0.745, p < 0.0001), which means that APC circulating levels are proportional to protein C levels and suggests that the protein C level is the limiting factor in the rate of protein C activation in vivo. PMID- 8713781 TI - A two-allele polymorphism in protein C inhibitor with varying frequencies in different ethnic populations. AB - cDNAs for protein C inhibitor (PCI), prepared from human liver RNA, contained two forms of PCI, designated PCI*A and PCI*B. While PCI*A is identical to the published PCI sequence, PCI*B differs in 4 of 1221 bp and two amino acids, A36V and K86E. Frequencies for the PCI*B allele, determined from genomic DNA, differed among ethnic groups. Frequency distribution and historical migration of modern man suggest that PCI*A arose from the PCI*B allele. Antigen levels in plasma homozygous for PCI*A or PCI*B equalled that of pooled normal plasma. K86E in PCI*B causes a charge alteration in helix D which is likely involved in heparin binding in antithrombin III but not likely involved in glycosaminoglycan binding in PCI. Kinetic studies showed that plasmas homozygous for PCI*A and PCI*B are similar in their APC inhibiting properties and in their heparin sensitivity, consistent with the idea that helix D in PCI is not involved in heparin binding. PMID- 8713782 TI - Mutations which introduce free cysteine residues in the Gla-domain of vitamin K dependent proteins result in the formation of complexes with alpha 1 microglobulin. AB - We have previously described a genetic factor IX variant (Cys18-->Arg) for which we demonstrated that it had formed a heterodimer with alpha 1-microglobulin through formation of a disulphide bond with the remaining free cysteine residue of the disrupted disulphide bond in the Gla-domain of factor IX. Recently, we observed a similar high molecular weight complex for a genetic protein C variant (Arg-1-->Cys). Both the factor IX and the protein C variants have a defect in the calcium induced conformation. In this study we show that the aminoterminus of this protein C variant is prolonged with one amino acid, cysteine. This protein C variant, as well as protein C variants with Arg9-->Cys and Ser12-->Cys mutations which also carry a free cysteine residue, are shown to be present in plasma as a complex with alpha 1-microglobulin. A prothrombin variant with a Tyr44-->Cys mutation, had not formed such a complex. Furthermore, complexes between normal vitamin K-dependent clotting factors and alpha 1-microglobulin were shown to be present in plasma at low concentrations. The data suggest that the presence of an unpaired cysteine residue in the propeptide or the N-terminal half of the Gla domain has strongly promoted the formation of a complex with alpha 1 microglobulin in the variants. PMID- 8713783 TI - Nafamostat mesilate, a broad spectrum protease inhibitor, modulates platelet, neutrophil and contact activation in simulated extracorporeal circulation. AB - Activation of humoral and cellular participants in inflammation enhances the risk of postoperative bleeding and multiple organ damage in cardiopulmonary bypass (CPB). We now compare the effects of heparin alone in combination with nafamostat mesilate (NM), a protease inhibitor with specificity of trypsin-like enzymes, in an extracorporeal circuit which simulates CPB. NM significantly inhibits the release of platelet beta-thromboglobulin (beta TG) at 60 and 120 min. Platelet counts do not differ. ADP-induced aggregation decreases in circuits with NM, which is due to a direct effect of NM on platelet function. NM prevents any significant release of neutrophil elastase; at 120 min, plasma elastase-alpha 1 antitrypsin complex is 0.16 micrograms/ml in the NM group and 1.24 micrograms/ml in the control group. NM completely inhibits formation of complexes of C1 inhibitor with kallikrein and FXIIa. NM does not alter markers of complement activation (C1-C1-inhibitor complex and C5b-9), or indicators of thrombin formation (F1.2). However, at 120 min, thrombin activity as measured by release of fibrinopeptide A is significantly decreased. The data indicate that complement activation during CPB correlates poorly with neutrophil activation and that either kallikrein or FXIIa or both may be more important agonists. The ability of NM to inhibit two important contact system proteins and platelet and neutrophil release raises the possibility of suppressing the inflammatory response during clinical CPB. PMID- 8713784 TI - Fat emulsion infusion potentiates coagulation activation during human endotoxemia. AB - Intravenous fat emulsions are frequently given to malnourished patients who are prone to suffer from infectious complications. As injection of low dose endotoxin represents a model to study the human response to acute infection, we sought to determine the effect of lipid emulsion infusion on endotoxin-induced activation of the hemostatic mechanism in man. Ten healthy men received a bolus intravenous injection of endotoxin (lot EC-5; 20 U/kg) midway through a 4-h infusion (125 ml/h) of either dextrose 5% (n = 5) or Intralipid 20% (n = 5). Lipid infusion potentiated endotoxin-induced coagulation activation, as indicated by higher plasma levels of the prothrombin fragment F1 + 2 and of thrombin-antithrombin III complexes (both p < 0.05 for the difference between groups). However, lipid infusion did not influence the fibrinolytic response to intravenous endotoxin, as reflected by similar increases in the levels of tissue-type plasminogen activator and plasmin-alpha 2-antiplasmin complexes in both groups. Endotoxin-induced appearance of plasminogen activator inhibitor type I was enhanced by lipid infusion (p < 0.05). These data suggest that fat emulsion infusion may enhance the tendency towards thrombotic complications in patients with infections. PMID- 8713785 TI - The importance of platelets in the expression of monocyte tissue factor antigen measured by a new whole blood flow cytometric assay. AB - Previous methods for the determination of monocyte tissue factor (TF) have been technically complex, difficult to standardize, prone to spuriously elevated results and difficult to implement in a clinical laboratory environment. We report the development of a two-color whole blood cytometric technique that overcomes many of these disadvantages. The assay uses small volumes of citrated blood (1.0 ml), can be performed in under one hour (if endotoxin stimulation is not performed), is reproducible (CV = 5%) and uses methodology commonly available in clinical laboratories. Baseline (mean +/- SD) expression of monocyte TF in normal subjects was very low (1.1 +/- 0.95%, Mean Fluorescence [Mean FL] 0.20 +/- 0.01) making relatively small increases easy to detect. Monocyte TF expression following endotoxin (LPS) stimulation for 1 h was 34.6 +/- 11.2% (Mean FL 0.32 +/ 0.04). LPS-stimulated activity varied between subjects (21-68%) but was remarkably consistent for individual subjects (CV = 5.4%). Stimulated monocyte TF expression was directly proportional to the platelet count and was reduced by platelet protective anticoagulants and by ingestion of aspirin. Non LPS stimulated monocyte TF was markedly increased, in a dose-dependent manner, by adding collagen to whole blood. This was apparently associated with platelet monocyte binding and could be abolished by anti-P-Selectin. We conclude that the whole blood flow cytometric assay of monocyte TF may be a valuable tool for clinical use and a useful model system for evaluating the humoral and cellular factors governing monocyte TF expression in a natural environment. PMID- 8713786 TI - A novel missense mutation in two families with congenital plasminogen deficiency: identification of an Ala675 to Thr675 substitution. AB - We used a polymerase chain reaction (PCR) strategy and restriction fragment polymorphism analysis to evaluate all 19 exons of the plasminogen (PLG) gene in a Japanese patient with congenital PLG deficiency and her family members (family C). Sequence analysis following amplification of each exon and its flanking regions showed a single G to A transition in exon 17, resulting in the conversion of an Ala675 codon (GCT) to Thr675 codon (ACT). Since this mutation generates a new Mae III site, the Mae III digestion patterns of the PCR-amplified exon 17 fragments from each family member were analyzed. In all cases, the patterns correlated with the activities and antigen levels of plasma PLG in those members. The identical G to A transition in the same codon of exon 17 was detected by a Mae III digestion experiment in another proband and her family members with congenital PLG deficiency (family K). Furthermore, 20 normal individuals examined had no Mae III restriction site at this location. We conclude that a G to A transition in exon 17 is responsible for the congenital PLG deficiency inherited in these two Japanese families. PMID- 8713787 TI - Platelet adhesion following the administration of tissue-type plasminogen activator and its reversal by vitamin E in rats. AB - Thrombolytic therapy is known to induce platelet-related side effects. We used a parallel-plate flow chamber, which was connected to the femoral artery of the rat, to measure platelet adhesion ex vivo. A collagen-coated arterioarterial shunt between two carotid arteries was used to measure shunt patency duration as an index of antithrombotic efficacy. Tissue-type plasminogen activator (t-PA), vitamin E, and the combination of these two were intravenously administered for 60 min. Measurements were performed before drug administration, and at 30, 60, 120 min after the initiation of drug infusion. Our results indicated that (1) treatments with t-PA or t-PA/vitamin E prolonged the time to shunt occlusion at 30 and 60 min; (2) t-PA enhanced platelet adhesion at 60 and 120 min; (3) vitamin E tended to reduce platelet adhesion; (4) t-PA/vitamin E reduced the t-PA enhanced platelet adhesion; (5) at the high-density area of platelet adhesion under t-PA treatment, the adherent platelets demonstrated severe morphological changes which could be blocked by vitamin E. These data suggest that t-PA may enhance platelet adhesion in rats and that this adverse effect can be suppressed by co-administration of vitamin E. PMID- 8713788 TI - Substitution of serine or threonine at position 473 of tissue-type plasminogen activator increases its stability in plasma. AB - Inactivation by slow acting inhibitors in plasma is of little consequence for thrombolysis with wild type t-PA, since it is rapidly cleared from the blood stream and constantly replenished through infusion. However, it becomes increasingly important as the clearance rate of t-PA is reduced, through mutagenesis, to enable the molecule to be long acting and administered by a single bolus injection. The substitution of serine for alanine at position 473 substantially reduced the slow inactivation that occurs at pharmacological levels of t-PA in plasma. Approximately 70% of the activity of A473S remained after 4 h incubation in human plasma compared to approximately 25% for wild type t-PA. Wild type t-PA and A473S showed the same stability in alpha-2-antiplasmin depleted plasma, indicating that the resistance of A473S to inactivation is a result of reduced reactivity towards alpha-2-antiplasmin, the primary slow acting inhibitor of t-PA. The second order rate constant for the inactivation of A473S by purified alpha-2-antiplasmin was approximately 4 fold less than that of wild type t-PA, which is consistent with the results obtained in plasma. Substitution of threonine at position 473 also produced inhibitor resistance, but glycine did not. The substitution of charged or bulky residues at position 473 destroyed enzymatic activity. The mechanism of inhibitor resistance for A473S and A473T appears to be a reduced reactivity towards substrates with arginine at the P1 position. The A473S mutation adds well to T103N, a mutation that causes an approximate 9 fold reduction in the clearance rate of t-PA. The double mutation variant, T103N, A473S had normal plasma clot lysis activity, and was stable in plasma over a 4 h incubation period at 37 degrees C in vitro. PMID- 8713789 TI - Fibrinolytic response to interferon-alpha in healthy human subjects. AB - Interferons (IFNs) are used for a variety of disorders. It has been postulated that part of the effects of IFN may be mediated by IFN-induced modulation of endothelial cells. Since the principal activating and inhibiting factors of the fibrinolytic system are synthesized and stored in endothelial cells, we have studied the effects on fibrinolysis and coagulation of the administration of recombinant IFN-alpha (5 x 10(6) U/m2) to healthy human subjects (n = 8) in a randomized controlled cross-over study. IFN-alpha significantly increased plasma levels of tissue-type plasminogen activator (t-PA) and urokinase-type plasminogen activator (u-PA). Simultaneously, plasma levels of the inhibitor of plasminogen activation, PAI-1, sharply increased. The net effect on plasma plasminogen activator activity (PA-activity) was a modest increase to 116% of baseline, however without a significant effect on plasmin generation, as reflected by plasma levels of plasmin-alpha 2-antiplasmin complexes. IFN-alpha had no effect on the plasma levels of thrombin-antithrombin III (TAT) complexes. We conclude that despite considerable effects on endothelial cells, IFN-alpha does not significantly alter the coagulant-fibrinolytic balance, although the occurrence of such changes under pathological circumstances is not excluded. PMID- 8713790 TI - Anti-thrombotic effect of a PAI-1 inhibitor in rats given endotoxin. AB - The aim of this study was to investigate the anti-thrombotic effects of an inhibitor of the plasminogen activator inhibitor-1 (PAI-1) in rats given endotoxin. In studies in vitro, PRAP-1, a Fab-fragment of a polyclonal antibody against human PAI-1, was shown to inhibit PAI-1 activity in rat plasma as well as to stimulate clot-lysis of the euglobulin fraction derived from rat plasma. Endotoxin administered to anaesthetised rats produced a marked increase in plasma PAI-1 activity. To study fibrin formation and lysis in vivo after intravenous (i.v.) injection of the coagulant enzyme batroxobin, 125I-fibrinogen was administered to the animals. The thrombi formed by batroxobin were rapidly lysed in control animals, while the rate of lysis was markedly attenuated in rats given endotoxin. PRAP-1 was administered i.v. (bolus+infusion) to rats given endotoxin and batroxobin and the PAI-1 inhibitor caused a dose-dependent decrease in the 125I-fibrin deposition in the lungs. An immunohistochemical technique was used to confirm this decrease in density of fibrin clots in the tissue. Furthermore, PRAP 1 decreased plasma PAI-1 activity in the rats and this reduction was correlated to the decrease in lung 125I-fibrin deposition at the corresponding time point. It is concluded that in this experimental model the PAI-1 antibody PRAP-1 may indeed inhibit thrombosis in animals exposed to endotoxin. PMID- 8713791 TI - Proteins of the fibrinolytic system in human thrombi. AB - The proteins of fibrinolysis have been quantified in human thrombi, to assess the balance between plasminogen activators and their major inhibitor PAI-1. The relative roles of PAI-1 and alpha 2-AP were also examined since we have previously shown that both platelet PAI-1 and plasma alpha 2-AP are important determinants of clot lysis in vitro. Extracts and sections were prepared from human thrombi for quantitative immunoassay and immunohistochemical staining respectively. PAI-1 and alpha 2-AP were present at high concentrations. Levels of t-PA and t-PA-PAI-1 complex were relatively low. Staining confirmed the presence of abundant PAI-1, associated primarily with platelet material within the thrombus and also with fibrin. Staining for alpha 2-AP was also intense and demonstrated strong association with fibrin. The alpha 2-AP concentration was similar to its high plasma concentration, whereas PAI-1 levels were up to 30 times greater than that in circulating blood, suggesting that active recruitment of platelets contributes to the high PAI-1 concentration in thrombi. PMID- 8713792 TI - Fibrin binding of plasminogen coated liposomes in vitro. AB - In this study, the fibrin binding properties of liposomes containing a number of plasminogen (Plg) molecules on the outside were compared to those of free (non liposomal) Plg in an in vitro model system. Fibrin monolayer coated 96-wells plates were used, containing fibrin monomer at a density of around 3.4 to 3.9 x 10(-4) nmol/cm2. These densities are similar to liposomal Plg-densities, thus allowing multivalent interactions to occur. In the panel of experimental conditions that was chosen, binding of free Plg and liposomes with Plg showed three main differences in characteristics. Firstly, in the fibrin binding of Plg liposomes not all Plg may be involved, but on the average 40% of the total amount of liposomal Plg. This was shown by lysing the liposomes after binding to the fibrin and estimation of truly bound Plg. With Plg-densities on the liposomes below the fibrin binding sites density, the maximal number of bound Plg molecules remains below the amount of available fibrin binding sites. Secondly, a higher binding rate by at least one order of magnitude was observed for liposomes with Plg compared to free Plg. Thirdly, liposomes with Plg exhibit a fibrin binding affinity which increases with Plg-density, because of the multivalent character of interaction. Liposomal Plg can successfully compete for fibrin binding sites with a 100 fold higher concentration of free Plg. These in vitro findings indicate that in view of avid and rapid fibrin binding, liposomes with attached plasminogen may be suitable for in vivo targeting to fibrin based thrombi. PMID- 8713793 TI - Quantitative comparison of fibrin degradation with plasmin, miniplasmin, neurophil leukocyte elastase and cathepsin G. AB - The relative contribution of plasmin, miniplasmin, PMN-elastase and cathepsin G to the fibrin-gel dissolution is studied. The global kcat/KM ratios are determined as a measure of the fibrinolytic catalytic efficiency using spectrophotometric kinetic analysis of the competition between fibrin and synthetic peptide substrates for the proteases, turbidimetric assay for fibrin dissolution and gel-filtration of the partially degraded fibrin. When the substrate is fibrin polymerized in the presence of 3 mM Ca2+, the value of this ratio is 4.3 x 10(5) M-1.s-1 for plasmin, 1.9 x 10(5) M-1.s-1 for miniplasmin, 5.0 x 10(4) M-1.s-1 for PMN-elastase and 2.2 x 10(3) M-1.s-1 for cathepsin G. When fibrin is polymerized without addition of Ca2+, the kcat/KM values are increased by a factor of 2.3 for plasmin, 2.0 for miniplasmin and 1.6 for cathepsin G, whereas that of PMN-elastase is unchanged. Progressive cross-linking of fibrin decreases the catalytic action of all studied proteases, but no change in their relative contribution to fibrinolysis is observed. When plasmin inhibitor (at physiological concentration) is also cross-linked to fibrin, the most efficient fibrinolytic enzymes are miniplasmin and PMN-elastase. The effect of 6-aminohexanoate on the formation of fibrin degradation products by plasmin and miniplasmin suggests that the high-affinity lysine binding site in the N terminal kringle domain of plasmin is involved in the interactions with the native polymerized fibrin, whereas the fifth kringle found in both enzymes participates in binding to newly exposed lysine residues. These results provide a quantitative basis for the evaluation of fibrinolytic efficiency and support the concept of synergistic fibrinolysis. PMID- 8713794 TI - Diabetes mellitus alters the effect of peptide and protein ligands on membrane fluidity of blood platelets. AB - The increased nonenzymatic glycosylation of platelet membrane proteins has been suggested to underlie platelet hypersensitivity in diabetes and the relationship of this to the reduced membrane lipid fluidity has been reported. As the modulation in membrane fluidity may determine the degree of accessibility of membrane receptors, the consequent alterations in membrane lipid-protein interactions in diabetes mellitus may also underlie the differentiated effects of various thrombotic and fibrinolytic agents on platelet membrane lipid bilayer. In the present study we employed electron paramagnetic resonance and fluorescence spectroscopy to explore the ligand-induced platelet membrane fluidity changes in diabetic state, i.e. under conditions when the membrane architecture is considerably altered. The yield of the excimer formation of pyrenemaleimide (PM), which depends directly upon the collisional rate and distances between molecules, was elevated in diabetic platelet membranes, thus pointing to the occurrence of some constraints in the structure/conformation of platelet membrane proteins in diabetes mellitus. Such an immobilization of PM was accompanied by the significant elevation in membrane protein glycation in diabetic platelets. The effects of various interacting ligands on platelet membrane fluidity were significantly lower in diabetic platelets, and the differences were much more distinct at the lower depths of a lipid bilayer. Nevertheless, the alterations in membrane lipid fluidity observed upon the interaction of a given ligand occurred with an approximately equal frequency in control and diabetic platelets. Moreover, the probability that these alterations were less profound in diabetic platelets was the same for all types of ligands studied. In diabetic patients the interaction of RGDS and tissue-type plasminogen activator with platelet membranes resulted in much smaller reductions of the h+/h0 parameters in 5-DOXYL-Ste acid labelled platelets, thus indicating a lesser rigidization of membrane lipid bilayer in diabetes. Likewise, the fluidizing effect of both fibrinogen itself and fibrinogen-derived peptides containing gamma-chain carboxy-terminal sequence H-12-V was less pronounced in diabetic platelet membranes. PMID- 8713796 TI - Promotion of the crosslinking of fibrin and alpha 2-antiplasmin by platelets. AB - Factor XIII (FXIII) is of high importance in the regulation of fibrinolysis. It crosslinks alpha 2-antiplasmin (alpha 2AP) and fibrin and by this way protects fibrin from the prompt elimination by plasmin. Although FXIII of platelets has been implicated in this protective mechanism, the role of platelets and platelet FXIII in the crosslinking process is far from being elucidated. As demonstrated by SDS PAGE and by immunoblotting for alpha 2AP, intact normal platelets resuspended in FXIII-free plasma or FXIII-free fibrinogen solution catalyzed the crosslinking of fibrin chains and also the crosslinking of alpha 2AP to fibrin alpha-chains. With FXIII-deficient platelets no crosslinking reaction could be observed indicating that the crosslinking with normal platelets was, indeed, due to platelet FXIII and not to another, putative platelet transglutaminase. However, the crosslinking of alpha 2AP to fibrin induced by the FXIII of intact platelets resuspended in FXIII-free plasma was considerably less extensive than the crosslinking carried out by the FXIII of normal plasma in the presence of FXIII-free platelets. Furthermore, the replacement of FXIII-free platelets by normal platelets in normal FXIII-containing plasma resulted in little, if any, difference in the crosslinking process. When crosslinking was induced by highly purified plasma FXIII the presence of intact FXIII-free platelets significantly accelerated the formation of alpha-chain polymers as well as the incorporation of alpha 2AP-fibrin alpha-chain hetero-dimer into these polymers. The results indicate that, in physiological conditions, platelet FXIII plays only a minor role in the crosslinking of alpha 2AP and fibrin; however, platelets, independently of their FXIII content, promote the crosslinking reaction by providing a catalytic surface on which the formation of highly crosslinked fibrin polymers is accelerated. PMID- 8713795 TI - Inhibition by Argatroban, a specific thrombin inhibitor, of platelet activation by fibrin clot-associated thrombin. AB - Clot-associated thrombin retains amidolytic activity, and is resistant to inhibition by heparin, but not to low molecular weight thrombin inhibitors. We show that clot-associated thrombin induces platelet aggregation, is resistant to heparin:antithrombin III, less so to recombinant hirudin (rHV2Lys47) but not to argatroban, an active-site directed thrombin inhibitor. Fibrin clots prepared with human fibrinogen and thrombin were used to aggregate rabbit washed platelets assessed by single platelet counting, thromboxane B2 (TXB2) immunoassay and scanning electron microscopy. Fibrin clots decreased platelet counts, and released TXB2. Electron microscopy showed platelet aggregates on the clot surface. Argatroban concentration-dependently inhibited such aggregation with IC50s of 21 nM and 13 nM versus aggregation and TXB2 release respectively. The IC50s of Argatroban against fluid-phase thrombin producing similar aggregation were 12 nM (aggregation) and 33 nM (TXB2). rHV2Lys47 was less active against clot induced aggregation (IC50 = 1.8 nM) than against fluid-phase thrombin (IC50 = 0.06 nM). Heparin had an IC50 of 0.02 mU/ml against aggregation induced by fluid phase thrombin, but much greater concentrations are required to inhibit clot induced aggregation (IC50 = 48 mU/ml). These data provide a basis for the superiority of direct-acting thrombin inhibitors over heparin in platelet rich thrombi. PMID- 8713798 TI - Epinephrine augments platelet recruitment to immobilized collagen in flowing blood--evidence for a von Willebrand factor-mediated mechanism. AB - Although elevated plasma epinephrine (epi) levels are associated with clinical atherothrombosis, the role of epi in platelet-vessel wall interaction has not been established. Our aim was to study the effect of high physiological epi (10 nM) in an experimental model which tests the interaction between platelets and immobilized collagen in whole blood. Shear forces and anticoagulation were modulated. Epi significantly enhanced platelet deposition, but only at high shear rate (1,600 s-1). In PPACK- or LMWH-anticoagulated blood, the increase in platelet deposition was 32 to 85% (p < 0.02-0.05). Furthermore, platelet aggregation was cotriggered with subthreshold concentrations of epi and ristocetin, and monoclonal antibodies against glycoprotein (GP) Ib (AN 51 and SZ 2) attenuated epi-induced aggregation. We conclude that epi is capable of augmenting platelet functions, which are dependent on the interaction of vWF with GP Ib and GP IIb/IIIa. Via this mechanism epi may promote arterial thrombosis in vivo. PMID- 8713797 TI - Specific cross-reaction of IgG anti-phospholipid antibody with platelet glycoprotein IIIa. AB - To study the pathological functions of anti-phospholipid (anti-PL) antibodies, we have analyzed their effect on platelet function. We identified an IgG anti-PL mAb, designated PSG3, which cross-reacted specifically with glycoprotein (GP) IIIa in human platelets and inhibited platelet aggregation. PSG3 bound also to certain polyanionic substances, such as double-stranded DNA, heparan sulfate, dextran sulfate and acetylated-LDL, but not to other polyanionic substances. The binding of PSG3 to GPIIIa was completely inhibited by heparan sulfate and dextran sulfate, indicating that PSG3 recognizes a particular array of negative charges expressed on both GPIIIa and the specified polyanionic substances. Since neither neuraminidase- nor endoglycopeptidase F-treatment of GPIIIa had any significant effect on the binding of PSG3, this array must be located within the amino acid sequence of GPIIIa but not in the carbohydrate moiety. Reduction of the disulfide bonds in GPIIIa greatly reduced its reactivity, suggesting that the negative charges in the epitope are arranged in a particular conformation. PSG3 inhibited platelet aggregation induced by either ADP or collagen, it also inhibited fibrinogen binding to activated platelets in a dose-dependent fashion. PSG3, however, did not inhibit the binding of GRGDSP peptide to activated platelets. These results suggest that the PSG3 epitope on GPIIIa contains a particular array of negative charges, and possibly affects the fibrinogen binding to GPIIb/IIIa complex necessary for platelet aggregation. PMID- 8713799 TI - Fibrinogen binding to ICAM-1 on EA.hy 926 endothelial cells is dependent on an intact cytoskeleton. AB - Fibrinogen is a ligand for Intercellular Adhesion Molecule-1 (ICAM-1), and enhances monocyte-endothelial cell interaction by coupling Mac-1 on monocytes to ICAM-1 on endothelial cells. We investigated the role of the cytoskeleton in fibrinogen binding to the human endothelial cell line EA.hy 926 using immunofluorescence techniques. In this cell line TNF alpha induced the simultaneous appearance of stress fibers and of ICAM-1, which was clustered predominantly on endothelial cell projections. Incubation of TNF alpha-stimulated endothelial cells with fibrinogen resulted in binding of fibrinogen to ICAM-1 on these cell projections. Disruption of the cytoskeleton by cytocholasin B abolished fibrinogen binding. Activation of protein kinase C with 12-O tetradecanoyl phorbol-13-acetate resulted in simultaneous loss of both stress fibers and fibrinogen binding. These results suggest that a connection between ICAM-1 and the cytoskeleton results in clustering of ICAM-1 on cell projections, which is required for fibrinogen binding. PMID- 8713800 TI - Heparin reverses the procoagulant properties of stimulated endothelial cells. AB - We examined the ability of unfractionated heparin to modulate the procoagulant activities of stimulated endothelial cells (EC). Confluent human venous umbilical EC were incubated with heparin before or after stimulation, then rinsed extensively to eliminate any heparin in the solution. EC, stimulated for 4 h with endotoxin and interleukin 1 beta, expressed tissue factor and prothrombinase activities. When EC were treated with heparin (6 and 60 micrograms/ml) during the last 10 min of the stimulation period, EC-related procoagulant activities were inhibited in a dose-dependent manner (80-90% inhibition at 60 micrograms/ml). The inhibition was antithrombin-dependent and it disappeared after heparin removal in less than 15 min at 37 degrees C but persisted at 4 degrees C. When EC were treated with heparin (60 micrograms/ml) for 24 h then extensively washed before stimulation, the anticoagulant effect was more modest (50% inhibition). The effect was antithrombin-dependent. Inhibition was maximum after 18-24 h of pretreatment of EC with heparin and was stable for at least 7 h. The cell surface displayed a "heparin-like" activity: treatment by heparin doubled the rate of thrombin-antithrombin complex formation and this effect was heparinase sensitive and chondroitinase ABC insensitive. Thus, heparin modulates the procoagulant properties of stimulated EC according to two distinct mechanisms. The first one is rapid and transient, probably related to the presence of heparin molecules bound at the membrane surface. The second is delayed and persistent, and our results suggest that it is mediated by an increase in the membrane heparan sulfate molecules. PMID- 8713801 TI - Primary binding domain of bovine von Willebrand factor fragment expressed in E. coli. AB - Bovine vWF cDNA has been cloned from a bovine endothelial cell library. A fragment of this cDNA, corresponding to amino acid sequence Leu 469-Ser 723, called primary adhesion domain (PAD-1), and containing the binding sites for platelet glycoprotein Ib (GPIb), heparin and collagen, has been expressed in E. coli. The reduced and alkylated form of fragment PAD-1 inhibited native vWF binding to GPIb. Fragment PAD-1 bound to heparin and botrocetin in a specific and dose dependent manner as did the native vWF. In a solid-phase assay, fragment PAD 1 bound to calf skin collagen in contrast to a human vWF recombinant fragment (Ser 445-Val 733) which was inactive in the same assay. The studies presented in this paper demonstrated that the A1 domain of bovine vWF contained the GPIb, heparin, botrocetin as well as collagen binding sites and that integrity of the disulfide bond (Cys 509-Cys 695), did not seem to be essential for binding of bovine vWF fragment to GPIb. PMID- 8713802 TI - The antithrombotic effect of aurin tricarboxylic acid in the guinea pig is not solely due to its interaction with the von Willebrand factor-GPIb axis. AB - Commercial aurin tricarboxylic acid (ATA) has been reported to interfere specifically with von Willebrand factor-glycoprotein Ib (vWF-GPIb) axis. This study was designed to explore the antithrombotic effects of ATA by examining its effects on guinea pig platelet function in vitro, in vivo and ex vivo. In vitro, addition of various concentrations of ATA to platelet-rich guinea pig plasma totally inhibited ristocetin-induced platelet aggregation, as expected. Unexpectedly, however, ATA similarly inhibited the aggregation induced by ADP, PAF, collagen, I-BOP (a thromboxane A2/prostaglandin H2 analogue) and arachidonic acid. In vivo, the antithrombotic action of ATA was assessed in a model of acute platelet-dependent guinea pig mesenteric artery thrombosis triggered by laser induced intimal injury. As the thrombotic response of arteries to such injury is a spontaneous cyclic recurrent process, 5 arteries in each animal were consecutively studied for 15 min each after i.v. bolus injection of 5, 7.5 or 10 mg/kg of ATA, which reduced the number of recurrent thrombi per artery in a dose dependent manner. The highest dose of 10 mg/kg induced maximal inhibition of thrombus formation (72%, p < 0.001) 5 min after injection. Ex vivo, platelet aggregation was assessed in blood samples taken before and after i.v. bolus injection of 10 or 15 mg/kg ATA. Ten mg/kg significantly inhibited collagen induced aggregation, and 15 mg/kg, the aggregation induced by ristocetin, ADP, PAF, collagen, I-BOP and arachidonic acid. The results of the in vivo studies confirmed that ATA is an effective antithrombotic agent. In the in vitro and ex vivo studies, ristocetin-induced platelet aggregation confirmed that ATA interacts with the vWF-GPIb axis, and suggests that the final common pathway of the aggregation induced by other agents tested consists of fibrinogen binding to the platelet GPIIb/IIIa receptor. We conclude that ATA interferes with vWF binding to GPIb, that it may interact with fibrinogen binding to GPIIb/IIIa, and that it might possess potent antithrombotic properties in platelet-mediated thrombosis. PMID- 8713803 TI - Heparin-induced thrombocytopenia: discrepancy between the presence of IgG cross reacting in vitro with fraxiparine and its successful clinical use. PMID- 8713804 TI - Autoimmune protein S deficiency and deep vein thrombosis after chickenpox. PMID- 8713829 TI - Remembering Robert J. Stoller (1924-1991). PMID- 8713830 TI - Notes on Foucault. PMID- 8713831 TI - The phenomenon of previewing and its contribution to the emerging sense of self. AB - Based on stringent measures of the infant's social interactions, Stern has postulated a four-step process of self-development. These internal factors contribute to the infant's emerging sense of the self as an entity separate and apart from others. There are, however, also significant external factors that contribute to the evolution of the self. The phenomenon of previewing provides infant with a means by which they develop, with the aid of an adaptive caregiver, their own abilities to achieve a sense of coherence and competence. The previewing process, combined with Stern's internal factors, provides a comprehensive theoretical basis for exploring how infants achieve selfhood. PMID- 8713832 TI - Psychoanalytic neonate models and non-Cartesian frameworks. PMID- 8713833 TI - Doubling back on Dennis Potter's doubles: the invisible man and the invisible woman. PMID- 8713834 TI - Masculinity and its vicissitudes: reflections on some gaps in the psychoanalytic theory of male identity formation. PMID- 8713835 TI - [The effect of 2',5'-oligoadenylates on the expression of the cellular proto oncogene c-myc during the development of Svec's leukemia in rats]. AB - The effects of 2',5'-oligoadenilates (2,5A) on the survival of Wistar rats inoculated with Svec leukemia cells and the proliferation of tumoral cells were investigated. An agreement between the expression of c-myc protooncogene and inhibition of leukemic cell proliferation by 2,5A was found. PMID- 8713837 TI - [Genotype identification by kappa-casein and BLAD mutation using the polymerase chain reaction in cattle. Bovine leukocyte adhesion deficiency]. AB - The investigation of BLAD mutation and genotypes for kappa-casein locus in groups of imported Holstein and Simmentals, as well as in Ukrainian Black-and-White cattle was carried out. The carriers of BLAD mutation were not detected. The frequencies of allelic B variant of kappa-casein (valuable trait associated with high the production of top quality cheese) were similar in both groups of imported cattle and substantially higher in Ukrainian cattle. PMID- 8713836 TI - [Alcohol dehydrogenase polymorphism and the genotypic adaptation of D. melanogaster to the action of selective factors]. AB - Analysis of experimental populations of Drosophila has shown different selective value of S- and F-alleles of Adh locus under selection for delayed aging, resistance to elevated temperature and ethanol. Selection for delayed aging and resistance to hyperthermia was accompanied by an increase of Adhs frequency in populations and genotypic adaptation to ethanol- by an increase of AdhF frequency. PMID- 8713838 TI - [Interbreed differences in cattle by "phenes"]. AB - Significant interbreed differences (between Simmentals and Black-and-White cattle) for 16 phenotypes of molar surface structure were detected in milk-cows. In Black-and-White cattle, 15 phenotypic variants of the head colour marks correlating with the body colour were found. PMID- 8713839 TI - [An analysis of the mutations in the mucoviscidosis gene: the origin and distribution of the delF508 major mutation in Ukraine]. AB - Seven mutations in CFTR gene have been screened in patients with cystic fibrosis from Ukraine. The frequency of major mutation delF508 among patients and healthy donors from different regions was studied. Data of age, origin and possible ways of spreading of this mutation in Ukraine have been discussing. PMID- 8713840 TI - [The status of the locomotor system in children living in localities with a high coefficient of inbreeding]. AB - Orthopedic examination of 1100 school-children was conducted in Rakhiv Region, Zakarpats'ka Oblast for detection of inbreeding. Significant correlation between the coefficient of inbreeding and the number of abnormalities in locomotor apparatus was detected. PMID- 8713841 TI - Basic DNA and RNA protocols. Introduction. PMID- 8713842 TI - The simultaneous isolation of RNA and DNA from tissues and cultured cells. PMID- 8713843 TI - Restriction endonuclease digestion of DNA. PMID- 8713844 TI - Agarose gel electrophoresis. PMID- 8713845 TI - Capillary blotting of agarose gels. PMID- 8713846 TI - Random primed 32P-labeling of DNA. PMID- 8713847 TI - Hybridization and competition hybridization of Southern blots. PMID- 8713848 TI - Utilization of DNA probes with digoxigenin-modified nucleotides in southern hybridizations. PMID- 8713849 TI - The preparation of fluorescein-labeled nucleic acid probes and their detection using alkaline phosphatase-catalyzed dioxetane chemiluminescence. PMID- 8713851 TI - The preparation of horseradish peroxidase-labeled nucleic acid probes and their detection using enhanced chemiluminescence. PMID- 8713850 TI - Monitoring incorporation of fluorescein into nucleic acid probes using a rapid labeling assay. PMID- 8713852 TI - 3'-end labeling of oligonucleotides with fluorescein-11-dUTP and enhanced chemiluminescent detection. PMID- 8713853 TI - The preparation of riboprobes. PMID- 8713854 TI - Native polyacrylamide gel electrophoresis. PMID- 8713855 TI - Use of silver staining to detect nucleic acids. PMID- 8713856 TI - End-labeling of DNA fragments. PMID- 8713857 TI - Northern blot analysis. PMID- 8713858 TI - RNA slot blotting. PMID- 8713859 TI - The RNase protection assay. PMID- 8713860 TI - Primer extension analysis of mRNA. PMID- 8713861 TI - S1 mapping using single-stranded DNA probes. PMID- 8713862 TI - Nonradioactive in situ hybridization for cells and tissues. PMID- 8713863 TI - In vitro packaging of DNA. PMID- 8713864 TI - Construction of mammalian genomic libraries using lambda replacement vectors. PMID- 8713865 TI - The production of double-stranded complementary DNA for use in making libraries. PMID- 8713866 TI - Construction of cDNA libraries. PMID- 8713867 TI - Screening lambda libraries. PMID- 8713868 TI - Subcloning strategies and protocols. PMID- 8713869 TI - Purification of DNA fragments from agarose gels using glass beads. PMID- 8713870 TI - Transformation of E. coli. PMID- 8713871 TI - Transformation of bacteria by electroporation. PMID- 8713872 TI - Preparation of plasmid DNA using alkaline lysis. PMID- 8713873 TI - The rapid boiling method for small-scale preparation of plasmid DNA. PMID- 8713874 TI - Plasmid preparations with diatomaceous earth. PMID- 8713875 TI - Polymerase chain reaction. PMID- 8713876 TI - Inverse polymerase chain reaction. PMID- 8713877 TI - Polymerase chain reaction with degenerate oligonucleotide primers to clone gene family members. PMID- 8713878 TI - Cloning PCR products using T-vectors. PMID- 8713879 TI - Direct radioactive labeling of polymerase chain reaction products. PMID- 8713880 TI - A rapid PCR-based colony screening protocol for cloned inserts. PMID- 8713881 TI - Use of polymerase chain reaction to screen phage libraries. PMID- 8713882 TI - Cloning into M13 bacteriophage vectors. PMID- 8713883 TI - Ordered deletions using exonuclease III. PMID- 8713884 TI - M13 phage growth and single-stranded DNA preparation. PMID- 8713885 TI - Preparation of ssDNA from phagemid vectors. PMID- 8713886 TI - A rapid plasmid purification method for dideoxy sequencing. PMID- 8713887 TI - DNA sequencing using sequenase version 2.0 T7 DNA polymerase. PMID- 8713888 TI - Pouring linear and buffer-gradient sequencing gels. PMID- 8713889 TI - Electrophoresis of sequence reaction samples. PMID- 8713890 TI - Direct sequencing of PCR products. PMID- 8713891 TI - Thermal cycle dideoxy DNA sequencing. PMID- 8713892 TI - Using the automated DNA sequencer. PMID- 8713893 TI - Terminal labeling of DNA for Maxam and Gilbert sequencing. PMID- 8713894 TI - DNA sequencing by the chemical method. PMID- 8713895 TI - Site-directed mutagenesis of double-stranded plasmids, domain substitution, and marker rescue by comutagenesis of restriction enzyme sites. PMID- 8713896 TI - A protocol for site-directed mutagenesis employing a uracil-containing phagemid template. PMID- 8713897 TI - In vitro translation of messenger RNA in a rabbit reticulocyte lysate cell-free system. PMID- 8713898 TI - In vitro translation of messenger RNA in a wheat germ extract cell-free system. PMID- 8713899 TI - One-step purification of recombinant proteins with the 6xHis tag and Ni-NTA resin. PMID- 8713900 TI - Structural allografting in revision total knee arthroplasty. AB - Between 1985 and 1991, 15 patients underwent structural allografting as part of revision total knee arthroplasty. All patients had large segmental, cavitary, or combination defects of the femur and/or tibia. Seven distal femurs and 12 proximal tibias required allografting. Patients were evaluated with physical examination, radiographs, and The Hospital for Special Surgery knee rating scale. Three patients died, leaving 15 allografts for follow-up study. The average age at surgery was 63 years. The follow-up period averaged 47 months (range, 30-101 months). Average range of motion before surgery was 4 degrees to 93 degrees, and after surgery, 2 degrees to 104 degrees. Average knee score was 47 before and 86 after surgery. Preoperative alignment averaged 5 degrees varus, ranging from 25 degrees valgus to 20 degrees varus, and postoperative alignment averaged 4 degrees valgus, ranging from neutral to 6 degrees valgus. All patients, except one, had improvement of pain and stability. All of the 15 allografts healed to host-bone and 13 showed evidence of incorporation. There were no infections or fractures of the allografts. One complication directly related to the allograft occurred; that patient had a tibial component fracture over a proximal tibial allograft 3 years after surgery. Three other complications occurred. One was tibial loosening in a patient who received a distal femoral allograft, the second was a proximal tibial fracture in a patient who received a distal femoral allograft, and the third was an intraoperative patellar tendon avulsion. These results suggest that structural allografting can provide a satisfactory method of managing large bone defects in the failed total knee arthroplasty. PMID- 8713901 TI - Outcome of the contralateral hip following total hip arthroplasty for osteoarthritis. AB - One thousand one hundred and sixteen patients who underwent unilateral total hip arthroplasty osteoarthritis between 1970 and 1980 were examined for development, progression, and total hip arthroplasty in the contralateral joint. At the time of the original surgery, 452 patients were diagnosed with bilateral osteoarthritis and 664 had a normal contralateral hip. The probability of osteoarthritis progressing in the contralateral joint is 78.6% at 10 years, with the chance of arthroplasty being 53.8%. The probability of a hip diagnosed as normal developing osteoarthritis is 36.5% at 10 years, with an 8.3% chance of requiring total hip arthroplasty. PMID- 8713902 TI - Uncemented press-fit total hip arthroplasty using the Identifit custom-molding technique. A prospective minimum 2-year follow-up study. AB - Fifty-three primary uncemented custom-molded Identifit (Depuy, Warsaw, IN) hip arthroplasties were evaluated prospectively at a mean follow-up period of 30 months. The custom technique provided the capability to reproduce the unique femoral offset, version, and height in each hip and to achieve high percentages of femoral canal fill. Surgical time for unilateral cases was a mean 153 minutes. Clinical results, however, were disappointing. Nine hips (17%) required stem revision for persistent thigh pain and limping. Of the remaining 44 hips, the mean Harris hip score was 83, and 20% experienced moderate to severe thigh pain and 50% had a limp. Radiographically, 65% of the stems had subsided and 27% had migrated into valgus. Survivorship analysis predicted an 80% stem survival rate at 43 months. A precise fit and fill of the femoral canal is not in itself sufficient for femoral implant stability in total hip arthroplasty surgery. PMID- 8713903 TI - A modified direct lateral approach for primary and revision total hip arthroplasty. A prospective analysis of 453 cases. AB - A prospective analysis of 453 consecutive primary and revision total hip arthroplasties (THAs) were reviewed. The modified direct lateral approach was used in 306 of 319 (96%) primary THAs and 115 of 134 (86%) revision cases for a total of 421 of 453 (93%) procedures. There were three dislocations in the revision group of patients. All three patients had at least two previous arthroplasties prior to their index surgery and at the time of surgery were noted to have very poor tissue quality, making the repair difficult. There were no postoperative dislocations in the primary THA group, with an overall dislocation rate of 0.7%. The incidence of postoperative limp in the primary THA group was 18%. The overall incidence, including revision surgery, was 27%. The presence of a persistent limp after initial rehabilitation was dependent on the number of previous surgeries, leg-length discrepancy greater than 2.7 cm, revision THA requiring a proximal femoral allograft, and preexisting deformities. Limp was therefore believed not to be related to the use of the modified direct lateral approach. PMID- 8713904 TI - Cemented total hip arthroplasty in patients with osteonecrosis. A 6-year minimum follow-up study of second-generation cement techniques. AB - Twenty patients with osteonecrosis of the femoral head underwent 28 total hip arthroplasties using cement from 1981 to 1985. Femoral reconstruction was by use of second-generation cement techniques. Twenty-four hips in 17 patients were available for review at a mean follow-up period of 7.7 years. The mean age at surgery was 55 years. Clinical evaluation demonstrated 79% excellent, 4% good, and 4% fair results. Three hips (12.5%) required revision for loosening. The cumulative probability of survival was estimated to be 85.7% at 10 years. Second generation cement techniques and implant designs did improve the clinical results in this high-risk group; however, the overall mechanical failure rate remained high. PMID- 8713905 TI - Inter- and intraobserver study of radiographic assessment of cemented total hip arthroplasties. AB - Radiographs are commonly used to identify loosened total hip prostheses. Interobserver and intraobserver variation was studied in an attempt to interpret 60 random pairs of radiographs with a time interval of 6 months to 12 years between the radiographs. Each of the four observers evaluated all pairs of radiographs independently, and the evaluations were repeated after 2 to 4 weeks. Each observer classified the femoral stem and the acetabular cup separately as stable or loose. Radiolucent lines of more than 2 mm were recorded. For the acetabular components, any change in inclination or migration was noted, and for the femoral components, subsidence or any change in varus or valgus of the stem was noted. There was considerable disagreement in observations of the acetabular component; agreement by all four observers was obtained in only 34 of 60 cases (57%). For the femoral component, agreement was obtained in 37 cases (62%). The observers agreed two and two (pairwise) on the acetabular component in 70 to 83% of cases and on the femoral component, in 72 to 82% of cases. Intraobserver variation was great with regard to the acetabular component, with kappa values ranging from 0.489 to 0.633. As to the femoral component, kappa values ranged from 0.737 to 0.800. Interpretation of radiographs of artificial hip arthroplasty is difficult, and comparisons between different series of implants should be made by the same person. PMID- 8713906 TI - Radiographic cost reduction strategy in total joint arthroplasty. A prospective analysis. AB - A consecutive series of 222 patients who underwent cemented total knee arthroplasty (124) and uncemented total hip arthroplasty (98) were evaluated prospectively. The purpose of this study was to determine if routine radiologic interpretation of postoperative total hip and total knee radiographs is cost effective. Also, the study was designed to determine if routine predischarge radiographs, in conjunction with recovery room radiographs, are worthwhile. There were no changes in postoperative patient management based on orthopaedic or radiologic review of either radiograph. No additional information was gained from review of the radiologic evaluations. Therefore, obtaining one series of routine inpatient postoperative total joint radiographs and eliminating postoperative radiologic consultation will significantly reduce costs without compromising patient care. PMID- 8713907 TI - Inpatient cost of primary total joint arthroplasty. AB - The financial and medical records of 98 patients undergoing either primary total hip or total knee arthroplasty were analyzed to determine the inpatient cost of their surgery. Eleven patients were excluded, leaving a total of 87 patients available for analysis who underwent 50 primary total hip and 37 primary total knee arthroplasties. Ten separate cost centers such as implant, anesthesia/operating room, nursing/hospital room, physical therapy, and laboratory were included in the evaluation. Correlations between total cost and demographic parameters such as patient sex, age, and length of stay were determined. Seventy-six percent of the inpatient cost of all total joint procedures consisted of implant, anesthesia/operating room, and nursing/hospital room costs. Reductions in the cost of these components represent a potential for cost containment. The strongest correlate for total cost was hospital length of stay (P < .0005). Unfortunately, there is evidence that length of stay has reached a plateau and cannot be reduced further without affecting the quality of care and the incidence of complications. PMID- 8713908 TI - Uncemented total hip arthroplasty using the CLS stem: a titanium alloy implant with a corundum blast finish. Results at a mean 6 years in a prospective study. AB - An uncemented titanium alloy stem with a corundum blast finish and an uncemented titanium fibermetal mesh socket were implanted in a series of 57 hips. These prostheses were selected for use in the youngest, most active, and/or heaviest candidates for total hip arthroplasty. Fifty hips were available for study at a minimum 60 months. At a mean 6 years, 92% of the hips were rated good or excellent. The mean Harris hip score was 92. One patient experienced mild thigh pain. The corundum blast finish was associated with reliable implant stability. Survival analysis predicted a 96% rate of implant survival at 92 months. Loss of bone density was rated mild, minimal, or none in 88% of the hips. Three hips developed severe bone loss due to systemic disease. Polyethylene wear was measurable in 86% of the hips. Twenty hips developed focal proximal femoral bone erosions. One hip had endosteal cavitation distal to zone 7. The presence of proximal femoral erosions or endosteal cavitation correlated positively with the presence of measurable polyethylene wear. The limited and proximal distribution of femoral bone erosion despite evidence of extensive polyethylene wear suggested that bone apposition to the corundum blast finish resulted in a barrier to migration of wear debris. PMID- 8713909 TI - Arthroscopic treatment of peripatellar fibrosis after total knee arthroplasty. AB - Forty-eight total knee arthroplasties (42 patients) were treated arthroscopically for symptomatic peripatellar fibrosis. All patients complained of knee pain before surgery. In addition, 47 knees (98%) had clicking or clunking, 35 (73%) had difficulty climbing stairs, and 9 (19%) had motion problems. The results of arthroscopy were 20 good (42%), 9 fair (19%), and 19 poor (40%). The mean follow up time was 32.8 months (range, 11-75 months). A fourth portal was required in 20 knees (17 patients, 42%), indicating the complexity and difficulty of debridement. No components were found to be loose at the time of arthroscopy. A constellation of symptoms are caused by peripatellar fibrosis. In contrast to past reports, the arthroscopic treatment of peripatellar fibrosis was found to be unpredictable. Debridement of the offending soft tissue did not necessarily guarantee a good result. Arthroscopic management of peripatellar fibrosis is recommended; however, limited and specific surgical goals should be established prior to intervention. PMID- 8713910 TI - Cementless hemispheric acetabular components. A 4- to 8-year follow-up report. AB - A retrospective radiographic analysis was performed on 71 cementless acetabular components followed from 4 to 8 years after surgery. A Harris-Galante (Zimmer, Warsaw, IN) cup was used in 46 cases and an Optifix (Smith & Nephew Richards, Memphis, TN) cup was used in 25 cases. There were 66 primary and 5 revision cases. Nonprogressive radiolucencies were common in both types of cups (64% of Optifix, 70% of Harris-Galante). Radiolucencies were almost exclusively less than or equal to 1 mm in width and were most common in zones 1 and 3. Ten cups had continuous but nonprogressive radiolucencies, none greater than 1 mm in all three zones. No radiolucent lines wider than 2 mm were seen in any case. Four cups had progressive radiolucency that stabilized. Radiolucency around fixation screws was seen in one case, and demonstrated stable ingrowth at revision surgery. No cases of osteolysis, screw breakage, migration, or loss of fixation surface occurred. A single case of a broken locking mechanism of a Harris-Galante cup 2 years after liner exchange is reported. PMID- 8713911 TI - Incidence of thigh pain after uncemented total hip arthroplasty as a function of femoral stem size. AB - Two hundred seventy-one patients (297 primary total hip arthroplasties) were evaluated for thigh pain 2 years after surgery. All femoral components were identical in wedge-shaped geometry but differed in size. All components were radiographically stable. Regression analysis revealed the presence of thigh pain to be directly correlated to increasing stem size (P = .014, r = .857). An even more significant positive correlation to thigh pain was present for proximal and distal component moments of inertia for bending in the mediolateral plane. Femoral stem size has a significant effect on the incidence of thigh pain. PMID- 8713912 TI - Accuracy of using radiographs for custom hip stem design. AB - If the shape of the femoral canal could be predicted with sufficient accuracy for the design of custom femoral stems, this would reduce the cost and provide a simpler design method than using computed tomography scans. Five groups of femurs were used for the study. The first two groups were used to determine an average femur shape, described by 25 transverse sections. In the next group of femurs, the shape was predicted from radiographs by distorting the shape of the average femur to conform to the radiographic outlines. The profiles were accurate to within 1 mm on average, the larger errors being in the trochanteric regions. In the proximomedial region, and in the distal canal, the accuracy was better than 0.3 +/- 0.8 mm. When femoral stems were designed from the actual and predicted canals, there was minimal difference in the sections of the two stems. The stems had a particular geometric form with a straight distal end, and flares at the proximomedial and proximoanterior locations; however, when stems designed by either method were inserted into canals prepared by distal reaming and proximal rasping, there was an error in fit in certain regions of approximately 1 mm. It was concluded that for femurs without serious abnormality of shape, a biplane radiographic method was sufficiently accurate for the prediction of canal shape and for the design of a standard type of custom uncemented stem. PMID- 8713913 TI - Long-duration metal-on-metal total hip arthroplasties with low wear of the articulating surfaces. AB - The 20-year performance of metal-on-metal hip articulations has not been reported. Five McKee-Farrar total hip prostheses and one Sivash prosthesis were obtained at revision surgery after a mean implantation time of 21.3 years. A radiographic, histologic, implant, and wear analysis was performed on these total hip implants with cobalt-chrome metal-on-metal articulations. All cases were associated with femoral component loosening, but the bearing surfaces performed remarkably well. The worst case estimate of combined femoral and acetabular linear wear was 4.2 microns per year, about 25 times less than that typically seen with polyethylene. Metal particles and foreign-body inflammation were seen in all cases, but the volume of reactive tissue was small compared with what is generally seen at revision of hips with a polyethylene acetabular bearing. This may be due to a reduced particle burden or a decreased inflammatory reaction to particulate metal, or both. In addition to articular wear, other sources of metal particles included femoral neck impingement on the acetabular rim, stem burnishing, and corrosion. Prosthetic hip reconstructions can fail for many reasons, including suboptimal femoral stem and/or acetabular cup design and/or fixation. By today's standards, the McKee-Farrar and Sivash stem and acetabular component designs are suboptimal; however, after more than 20 years of use, the metal-on-metal bearing surfaces in these cases demonstrated low wear and do not appear to be the cause of failure. Recent advances in total hip arthroplasty, which include improved implant design, materials, manufacturing, and fixation, combined with a better understanding of the mechanisms of implant loosening and failure, suggest that the cobalt-chrome metal-on-metal bearing be reexamined as an alternative to polyethylene when exceptional durability is required. PMID- 8713914 TI - Modularity in primary total hip arthroplasty. PMID- 8713915 TI - Modularity is unnecessary in primary femoral THA but has some advantages in primary acetabular THA. PMID- 8713916 TI - Management of the chronic irreducible patellar dislocation in total knee arthroplasty. AB - Neglected dislocation of the patella with gonarthrosis, genu valgum, flexion, and external rotation deformity is rarely encountered. Experience with five total knee arthroplasties in three patients with chronic patellar dislocation and gonarthrosis is reported. All knees had a modified proximal patellar realignment and arthroplasty with a constrained prosthesis. Preoperative Hospital for Special Surgery knee scores averaged 55. Average follow-up period was 40 months. At latest follow-up examination, the average Hospital for Special Surgery knee score was 83, the Knee Society knee score was 95, and the functional score averaged 50. There was one complication: a full-thickness lateral skin necrosis requiring flap coverage. The patellar score was zero in all knees. Four knees had mild quadriceps weakness. Three knees rated as excellent and two as good on both The Hospital for Special Surgery and Knee Society rating systems. Radiographic analysis revealed no radiolucent lines or osteolysis. The patellas were centralized in the trochlear groove in all patients. Patellar height averaged 14 mm (range, 12-17 mm). In conclusion, satisfactory results were obtained by restoring axial alignment with a constrained implant and realigning the patella with an extensive proximal realignment. PMID- 8713917 TI - Occlusion of the external iliac vein by cement. AB - The case of an 80-year-old woman who sustained occlusion of the external iliac vein by methyl methacrylate cement following total hip arthroplasty is presented. The cement was removed via a pelvic approach without the need for revision of the acetabular cup and led to resolution of her symptoms. PMID- 8713918 TI - Femoral artery and nerve compression by bulk allograft used for acetabular reconstruction. An unreported complication. AB - Absent pulses in the ipsilateral limb and a femoral nerve palsy occurred after revision total hip arthroplasty. Emergent arteriogram revealed an occlusion of the femoral artery inferior to the inguinal ligament. The femoral artery was found to be compressed by the bulk allograft bone used for reconstruction of an anterior segmental acetabular defect. No laceration or thrombosis of the artery was present. A high-speed burr was used to remove the anterior portion of the offending allograft. Pulses were immediately restored and the patient had no long term vascular sequelae. The femoral nerve palsy has slowly resolved over 2 years. PMID- 8713919 TI - Early failure of Hylamer acetabular inserts due to eccentric wear. AB - The authors performed a retrospective review of the survivorship of Hylamer (DePuy Dupont Orthopaedics, Warsaw, IN) acetabular liners following primary arthroplasty of the acetabulum. Between 1990 and 1992, 193 total hip arthroplasties were performed in which Hylamer acetabular liners were implanted. One hundred forty-three of these hips (74%) were available for clinical and radio graphic follow-up evaluation after a minimum of 2 years. The mean follow-up period was 2.5 years. Five Hylamer liners have been revised for eccentric wear at a mean of 37 months postimplantation and one additional liner is scheduled to be revised at the 34-month follow-up evaluation (total, 4.2% failure rate due to wear). Survivorship of the Hylamer liner, with failure defined as revision due to eccentric wear, was calculated to be 86% (SE, 5.5%) at 4 years. The authors have discontinued the use of Hylamer acetabular liners because of this high early failure rate. PMID- 8713920 TI - Levels of lactate dehydrogenase in osteoarthritic and failed total knee joints. AB - Synoval fluid lactate dehydrogenase may be used as a marker of joint inflammation. Macrophage phagocytosis may result in cell llysis and release of intracellular lactate dehydrogenase. This study demonstrates that synovial lactate dehydrogenase levels are markedly increased in failed total knee arthroplasty as compared to end-state osteoarthritic knee joints. PMID- 8713922 TI - Managing acetabular defects. PMID- 8713921 TI - Postoperative blood salvage and reinfusion in total knee arthroplasty. PMID- 8713923 TI - Tear film changes associated with normal aging. AB - We examined a group of normal subjects to evaluate the tear film function by measuring reflex tear capacity using the Schirmer test without anesthetic, steady state tear flow using fluorophotometry, meibomian gland function based on gland drop-out, expressible lipid volume and viscosity, tear loss from evaporation, and tear osmolarity. We found the average tear flow to be 0.19 +/- 0.19 microliters/min, lower than previously measured. The average tear volume was also correspondingly less, 2.74 +/- 2.0 microliters. We found there was a decline associated with advancing age for all parameters except the decay constant and lipid volume. The highest correlations with aging were tear volume (-0.64), Schirmer test (-0.63), and tear osmolarity (0.59). Our results indicate that there is a decline in tear film function throughout life and that tear flow is considerably less than previously estimated. PMID- 8713924 TI - Decrease in corneal sensitivity and change in tear function in dry eye. AB - Our purpose was to investigate the relationship between insufficient tear secretion and decreased corneal sensitivity. We studied 59 patients with dry eye, 15 of whom had Sjogren's syndrome (SS), and 26 healthy subjects. Corneal sensitivity was measured by the esthesiometer of Cochet and Bonnet. Schirmer test with or without anesthesia, tear clearance rate, tear function index, and rose bengal and fluorescein staining were also evaluated. The mean corneal sensitivity of either dry-eye group (4.6 +/- 1.2 and 4.5 +/- 1.2 cm for non-SS and SS dry eye, respectively) was significantly lower than that of the control (5.8 +/- 0.4 cm, p < 0.001). Corneal sensitivity correlated significantly with the Schirmer values with anesthesia and the tear function index in the two dry-eye groups and the control (p < 0.05). There were significant relationships between corneal sensitivity and the rose bengal and fluorescein scores in the three groups (p < 0.05). Hyposecretion of tears in dry eye may lead to pathologic changes in corneal epithelium and a decline in corneal sensitivity. Prompt treatment of dry eye is essential to maintain a normal corneal protective mechanism. PMID- 8713925 TI - The effect of osmolality on the shedding rate of the corneal epithelium. AB - It has been stated that in dry eye hyperosmolality damages the epithelium of the ocular surface by increasing the rate at which cells are shed. To test this hypothesis, paired excised rabbit corneas were superfused with balanced salt solutions in which all conditions were held constant except the osmolality in contact with the epithelium. For a period of 400 min the epithelium of one of the corneas was exposed to one of seven test osmolalities (200-425 mOsm/kg), whereas the other cornea was used as a control (305 mOsm/kg). The number of cells shed from each corneal surface was counted, and thickness changes in the epithelium and stroma were measured. Only hypoosmotic solutions of 260 mOsm/kg or less increased the number of cells shed relative to the control. None of the hyperosmotic solutions significantly increased the shedding rate. There was no significant change in the thickness of the epithelium, whereas the stroma swelled in hypoosmotic solutions and thinned in hyperosmotic solutions. It is concluded that hyperosmolality in the range encountered in dry eyes is not sufficient in itself to increase the shedding rate. PMID- 8713926 TI - Effect of pharmacological dilation and constriction of pupil on corneal topography. AB - A prospective study was undertaken to investigate the effect of pharmacological pupillary dilation and constriction on corneal topography. Corneal topography was performed using the EyeSys system on 28 normal subjects before and after pupil dilation, constriction, and then with the pupil intact (no medication). Our results show that both flattening and steepening of the corneal curvature were found in all three examination periods. The numbers of corneas with flattening and steepening in each examination period were compared and found not to be statistically different. Mean change of corneal flattening was compared with mean change of corneal steepening in each examination period and again no significant difference was found. These results indicate that the slight changes in corneal power and radius following pharmacological pupillary dilation or constriction may be due to either the inability to repeat by the subjects, or error by observers or instrument. PMID- 8713927 TI - Intrastromal epithelial accretion follows deep excimer annular keratectomy. AB - The purpose of this study was to evaluate corneal reepithelialization and wound healing following annular excimer keratectomy. Two sets of experiments were performed on 35 rabbit eyes. In the first set of experiments, experiment I, deep Fresnel excimer keratectomy was performed, with a 6-mm outer and 3-mm inner diameter. Animals were sacrificed at 1, 5, 12, and 16 weeks, and corneas were examined by light and electron microscopy. In experiment II, the central epithelium was left intact, and superficial and deep mid-peripheral excimer annular keratectomies were performed measuring 6 mm in outer and 3 mm in inner diameter. Animals were sacrificed at 1 week, and corneas were examined by light microscopy. Following deep Fresnel excimer keratectomy (experiment I), corneas showed stromal edema in the central 3-mm zone. Intrastromal islands of epithelial cells with PAS positive basement membrane-like structures were seen histologically at 1 week. Electron microscopy showed loss of stromal collagen in areas adjacent to epithelial islands; in areas distant from the epithelial islands, the stromal collagen appeared normal. The overlying central stroma sloughed after 5 weeks. Anterior stromal scarring was observed. In experiment II (mid-peripheral annular keratectomy), intrastromal epithelial accretion was present in corneas with deep annular keratectomy but not in superficial annular keratectomy. Intrastromal epithelial accretion follows deep excimer annular keratectomy and is associated with adjacent stromal degradation. PMID- 8713928 TI - Inefficacy of collagen shields in the rabbit corneal wound-healing process. AB - A controlled study in the rabbit eye was performed to clarify the suitability of collagen shields in accelerating the wound healing process in damaged cornea. After a standardized bilateral keratectomy, the spontaneous evolution of the healing process in the right eye (control eye) was compared with the evolution in the left eye, treated with a collagen shield (a shield of the 12- or 24-h type). The healing area was measured by planimetry after fluorescein staining at 0, 24, 48, and 72 h after keratectomy. Histological and immunohistochemical analysis of the healing process was also performed. Reepithelization of the damaged cornea was almost complete at 72 h, and there were no differences in the time course of the healing process between control and treated eyes. There was an important polymorphonuclear infiltration in treated eyes, mainly composed of eosinophiles, which was not present in control eyes. This indicates a subacute inflammatory immunological reaction. It is concluded that the usefulness of collagen shields should be reappraised, especially in injured corneas. PMID- 8713929 TI - Corneal wound healing in the rabbit eye: activity of a fibronectin-splitting product. AB - The corneal wound-healing properties of fibronectin (FN) and a fibronectin hydrolysate (FNH) have been evaluated in comparison with commonly used drugs. Nonpenetrating bilateral surgical keratectomy was performed in male albino rabbits. The left eye was treated with the active product, whereas the right eye served as a control (vehicle). The healing area was measured by planimetry after fluorescein staining at 0, 24, 48, and 72 h after keratectomy. Histological and immunohistochemical analysis of the healing process was also performed. Results were as follows: (a) Nandrolone (p < 0.005) and asiaticoside (p < 0.001), both at 10 mg/ml, in eyedrops delayed the healing process. (b) An ointment containing vitamin A and amino acids also delayed the process but at the limit of statistical significance (p = 0.055). (c) FNH (20-80 mg prot/ml) significantly improved the quality and shortened the time of the healing process at 60 mg prot/ml and above. (d) Human FN (100-800 micrograms/ml) did not affect the healing process. Immunohistochemical analysis demonstrated that FNH accelerated the appearance of endogenous FN in the damaged cornea earlier than in the control eyes. It is concluded that FNH may be useful in the management of corneal wounds, whereas the effectiveness of FN is doubtful. PMID- 8713930 TI - The Topcon SP 1000 and Image-NET systems. A comparison of four methods for evaluating corneal endothelial cell density. AB - The objective of this study was to compare four methods of counting corneal endothelial cell density using the Topcon SP 1000 microscope and Image-NET digital graphic software. Two independent observers quantified cell density from 63 endothelial photomicrographs using the various standardized techniques. The first method was that suggested for use with Topcon SP 1000, employing five different reference grids to compare subjectively with the endothelial cell density of the unknown sample. The other three methods involved the automated, semiautomated, and manual procedures developed for use with the Image-NET system software. The manual Image-NET system is presently considered to yield accurate cell morphology data. Confidence limits and standard errors of mean differences between values obtained by different methods were used to evaluate agreement and reproducibility of computerized methods. Sensitivity and specificity were calculated for two different threshold limits of endothelial cell density. Results showed that the automated Image-NET system is not reliable for clinical use because of its poor agreement with other methods and its lack of sensitivity and specificity at the selected threshold limits of endothelial cell density. The comparative Topcon method of using reference grids, although inexpensive and accurate for 500 cell/mm2 increments, was considered too imprecise for most clinical situations. The semiautomated Image-NET system, in half the analysis time required by the manual method, provided endothelial cell count estimates that were not clinically different from those obtained with manual counting. PMID- 8713931 TI - A numerical test of the normal incidence uniaxial model of corneal birefringence. AB - The suggestion that the central cornea can be modeled as a uniaxial birefringent material with its optic axis normal to the surface is explicitly tested by numerical calculations. A theoretical framework is presented to model the corneal stroma as a series of stacked, uniaxial birefringent layers (lamellae). Calculations are then made of the transmission of normally incident, linearly polarized light through model systems having various azimuthal orientations of the layers, motivated by the suggestion of an overall "random" organization of the stromal lamellae. It is concluded that the uniaxial description, and the assumptions upon which that description is based, do not hold for the cornea. In particular, the calculations are in agreement with recent experiments in which one always observes a non-zero cross-polarized transmission (hence birefringence) at normal incidence. PMID- 8713932 TI - The immunohistochemical composition of the human corneal basement membrane. AB - Frozen sections of human cornea and conjunctiva were stained by immunofluorescence and immunoperoxidase methods with a panel of antibodies against various basement membrane (BM) and extracellular matrix proteins to study the composition of the human corneal BM. Laminin (Ln) immunoreactivity was detected in the BMs of the blood vessels and conjunctival, limbal, and corneal epithelia by using a polyclonal antiserum and monoclonal antibodies (MAbs) against Ln chains alpha 1, beta 1, and gamma 1, but in the cornea the reaction with MAbs against alpha 1, beta 1, and gamma 1 chains was clearly weaker than in the other BMs and in some sections totally absent. Immunoreactivity to Ln alpha 2 chain was detected only in the BM of the limbus, whereas Ln beta 2 chain was detected in the BMs of blood vessels and that of conjunctival and limbal epithelia. Ln-5 and Ln alpha 3 and beta 3 chains were seen in the BMs of the corneal, limbal, and conjunctival epithelia coaligning the integrin beta 4. Immunoreactivity for collagen type IV was lacking in the BM of the corneal epithelium, but present in the BMs of the limbal and conjunctival epithelia as well as in the vascular BM. Collagen type VII immunoreactivity was seen in the BM areas of the corneal, limbal, and conjunctival epithelia. BMs of corneal, limbal, conjunctival, and vascular epithelia expressed fibronectin (Fn), including extradomain A cellular Fn and oncofetal cellular Fn isoforms, all lacking, however, extradomain B cellular Fn. The present results give new information on the Ln chain distribution and show that the BM of the corneal epithelium differs from that of surrounding epithelia by the lack of collagen type IV. PMID- 8713933 TI - Chronic localized conjunctival chemosis. AB - Conjunctival chemosis is a commonly encountered clinical finding stemming from the presence of excess fluid in the conjunctiva. It is typically self-limiting or reversible if the underlying condition is treated. The authors present a series of seven cases of chronic localized conjunctival chemosis. Each patient had a localized area of dependent conjunctival edema for >or= 6 months. Evaluation of each patient included clinical examination, laboratory studies, and neuroimaging to attempt to elucidate the pathogenesis of the chemosis. Conjunctival biopsy was performed in six of the seven patients. In all of the patients studied, clinical examination uncovered no definitive signs of local inflammation. Laboratory evaluation was normal, and neuroimaging failed to confirm obstruction of venous or lymphatic drainage. Conjunctival biopsies showed chronic tissue inflammation or lymphangiectasia. The diagnosis of chronic localized conjunctival chemosis (CLCC) can be made if localized conjunctival edema persists for 6 months. Evaluation of biopsy specimens supports the theory that CLCC may result from scarring and structural alteration of conjunctival lymphatics, which appears to irreversibly affect the distribution of fluid in the conjunctiva. PMID- 8713934 TI - Biofilm formation in infectious crystalline keratopathy due to Candida albicans. AB - Infectious crystalline keratopathy (ICK) is an uncommon, indolent corneal infection in which the slow clinical course contrasts with the rapid laboratory growth and microbiological sensitivities of the infecting organism. This prospective study aimed to determine whether biofilm production was the cause of this disparity. A case of failed medical management of ICK in a patient with Stevens-Johnson syndrome is presented. A penetrating keratoplasty yielded corneal tissue that was freshly fixed for electron microscopy using 0.05% ruthenium red and 2.5% gluteraldehyde. Candida albicans was grown from 3/3 broths, and fungi with morphology consistent with Candida were seen on histological examination. Electron microscopy revealed microorganisms morphologically typical of Candida surrounded by a polysaccharide-rich glycocalyx consistent with a biofilm. We concluded that Candida albicans is capable of producing a biofilm and is a known cause of ICK. This case is supportive evidence that biofilm production is associated with cases of ICK and may explain the chronic, pauciinflammatory features of ICK and its relative resistance to antibiotic treatment. PMID- 8713935 TI - Acquired immunodeficiency syndrome and ocular calcification. AB - We report the presence of peculiar nonbanded calcification of the cornea in three patients with acquired immunodeficiency syndrome, discovered on postmortem histological examination of the globes. In one patient, multiple calcium salt precipitates also affected other ocular and orbital structures. Calcium deposits were present in the corneal stroma but spared Bowman's layer, as is usually seen in primary and secondary corneal calcification. The calcified areas were positive to von Kossa, alizarin red, alcian blue, and colloidal iron stains. Electron probe analysis of the three cases showed the presence of calcium and phosphorus in a ratio characteristic for hydroxyapatite. No predisposing factors could be found. The possible role of associated alterations in the mucopolysaccharide content or composition in the calcified areas is unclear. PMID- 8713936 TI - Superior limbic keratoconjunctivitis in association with scarring of the superior tarsal conjunctiva. AB - Three patients are reported with clinical findings compatible with a diagnosis of superior limbic keratoconjunctivitis (SLK) that developed in association with scarring of the palpebral conjunctiva. Although the etiology of SLK is unknown, these patients support the theory of a mechanical cause for SLK. PMID- 8713937 TI - Co-occurring adenoviral keratitis and hemorrhagic cystitis. A case report. AB - Adenoviruses are ubiquitous viruses, commonly causing ocular infections. They are also known to cause multisystem infections. External ocular infections caused by adenovirus are most often due to contaminated secretions. We report a patient in whom genitourinary symptoms developed and subsided along with ocular symptoms. Adenovirus was isolated on ocular culture. This is the first reported case of concurrent adenoviral keratitis and hemorrhagic cystitis. Eliciting a history of genitourinary and respiratory symptoms is important in patients with ocular adenoviral infections. PMID- 8713938 TI - A histochemical and x-ray diffraction study of keratoconus epikeratoplasty. Report of two cases. AB - Two epikeratoplasty lenticules (Epi-L) with keratoconus host corneas (Epi-H) were removed by penetrating keratoplasty because of poor visual acuity although they were clinically clear. Light microscopy showed that the keratocytes were underpopulated and unevenly distributed in Epi-L. Transmission electron microscopy showed that abnormal sized proteoglycans (PGs) were unevenly accumulated in the stromas of both Epi-L and Epi-H. The low-angle x-ray diffraction results from case 1 showed lower collagen interfibrillar spacings, compared with those from one keratoconus cornea, seven cryolenticules, and four normal human corneas. The fibril diameters and the collagen axial periodicity D were similar in all tissues. Allowing selected samples to swell, the interfibrillar spacing increased by 30.6-45.5% in Epi-H and by 29.8% in the keratoconus control. However, it increased by only 7.6-9.7% in Epi-L and by 11.4% in a cryolenticule. The uneven distribution of PGs and the lower interfibrillar spacings in the swollen lenticules suggest that there is an uneven distribution of water in different parts of the Epi-L and Epi-H stromas. This may lead to a variation of the refractive index or optical properties in the stroma and the deterioration of the visual image, which may explain the poor visual results. PMID- 8713939 TI - Perfluorodecalin corneal toxicity: five case reports. AB - Perfluorodecalin is a perfluorocarbon liquid used intraoperatively in retinal detachment repair. It is usually removed at the end of the procedure; however, residual amounts may be retained when poor corneal clarity or intraocular hemorrhage obscures the view. No clinical reports exist on the consequences of retained perfluorodecalin in the anterior segment. We report five cases in which perfluorodecalin was in prolonged contact with the cornea. The period of time for corneal pathology to occur and the role perfluorodecalin played in the etiology of such changes is discussed. A total of 348 patients with retinal detachments in one retinal practice underwent repair using pars plana vitrectomy combined with intraoperative perfluorodecalin between January 1992 and May 1994. Postoperatively, residual perfluorodecalin was observed in the anterior chamber in contact with the corneal endothelium in five patients. The patients were followed clinically for a period of up to 18 months. Four of five patients developed corneal changes from prolonged contact with perfluorodecalin. Corneal edema developed in the area perfluorodecalin-endothelial contact in three of five eyes. The period of perfluorodecalin-endothelial contact before corneal decompensation occurred ranged from 4 to 13 weeks. Two eyes required penetrating keratoplasties for progressive corneal edema. Corneal edema was reversed in one eye after removal of perfluorodecalin from the anterior chamber via multiple paracentesis. One of the remaining eyes developed deep corneal vascularization without edema in the area of perfluorodecalin contact after 12 months. These observations suggest that corneal toxicity may be induced by intraocular perfluorodecalin if it is allowed direct contact with the corneal endothelium for periods as short as 1 month. Some of these changes may be reversible if perfluorodecalin is aspirated from the anterior chamber. Further investigations are required to examine perfluorodecalin-induced corneal toxicity. PMID- 8713940 TI - [Dental complications of radiotherapy of tumors of the nasal cavity in childhood]. AB - Authors have presented a case-report of a child with no or hardly developed roots of permanent teeth in the maxilla. Histological examinations of the removed teeth showed amelogenesis imperfecta and dentinogenesis imperfecta. The disturbance in root development is considered to be the consequence of the radiological treatment which was given to the child at the age of four because of the endodermal sinus tumor of the left middle nasal passage. PMID- 8713941 TI - [Clinical comparison of the calculus inhibiting effect of three commercially available toothpastes]. AB - A twelve-week independent and double-blind clinical study was conducted on a sample of 143 calculus forming adult male and female subjects--with the average age of 39.11 years--to compare directly the anticalculus efficacy of three commercially-available dentifrices, as compared to a placebo dentifrice. The three commercially-available dentifrices were: Colgate Total toothpaste, Colgate Tartar Control toothpaste and Pepsodent Ultra toothpaste. All these three dentifrices provided statistically significant reductions in supragingival calculus formation, as compared to a placebo dentifrice. There was no statistically significant difference among the three commercially-available dentifrices with regard to anticalculus efficacy. PMID- 8713942 TI - [The use of Dormicum in general anesthesia for oral surgery]. AB - This paper reports on the use of Dormicum for anaesthetization in 385 oral surgical operations on patients older than 45 years. These patients included high proportions of heavy smokers and heavy drinkers. These habits not only play an important role in the development of the basic disease, but also lead to diseases of the heart, vascular system, lungs and liver. Dormicum was applied for premedication and to introduce and maintain anaesthesia. The changes occurring in the circulatory and respiratory parameters in response to the drug were not significant clinically. Dormicum is rated as an excellent agent for the anaesthetization of high-risk oral surgical patients. PMID- 8713943 TI - [Current trends in antibiotic therapy in dentistry]. AB - The place and role of surgical, dental physio- and pharmacotherapy in oral and head and neck diseases has been debated for decades. In addition the price and reimbursement system had been changed lately in our country that also underlines the relevance of this issue. The overconsumption (1.7 packages per inhabitant per year) of antibiotics is also proven in Hungary. Beside surgical and dental interventions the treatment of dental inflammations caused by pathogenics requires the administration of different kinds of antibiotics. The first choice antibiotic is determined on the base of clinical features and general resistance of oral flora in the given period. When the first choice antibiotic proves to be ineffective, treatment has to be followed by antibiotics determined on individual resistance. In stomatology antibiotics are administered for prevention or treatment of dental inflammations. The length of therapy has a great importance which is usually 4 days in prevention and 5 days in treatment of dental inflammations. The modern antibiotics have to comply with the following requirements: efficacy, safety and cost-effectiveness. The typical mistakes in the administration of antibiotics can be avoided by the application of therapeutical schedules. The macrolide and cephalosporin derivates have become more and more popular. The mixed bacterium flora has been changing in time and also influenced by geographical factors depending on the habit of prescriptions. Therefore the experiences obtained in other countries are not applicable. PMID- 8713944 TI - [Comparative evaluation of root canal enlarging instruments studied on extracted human teeth. II. Micro-motor operated instruments]. PMID- 8713945 TI - [Esthetic and social indications for the "redressement force" technic and the hazards of this surgical procedure]. AB - The authors report on-, and evaluate the type of operation known as "redressement force" for the correction of retained upper canines. The risk of the surgical intervention is extremely high, but there are good reasons for the indications, too. PMID- 8713947 TI - Strategies for cell biological studies in oxytocinergic neurons. AB - Hypothalamic magnocellular neurons expressing the neurohypophysial peptides, oxytocin (OT) and vasopressin (AVP), have historically served as valuable models for cell biological studies of peptidergic neurons. This is because of the relative ease with which the perikarya, axons, and nerve terminals of these cells can be isolated for anatomical, biochemical, and physiological analysis. A popular strategy is to harness the regulatory elements in the OT and AVP genes in order to target specific molecules to these cells in vivo, so as to elucidate various cell biological issues, e.g., pacemaker, prohormone processing, axonal transport, and secretion mechanisms. The peptide genes are good sources for regulatory controls since they are relatively abundantly and specifically expressed in these neurons in the CNS. The major problem has been to identify the critical regulatory elements in these genes which are responsible for the cell specific expression. Recent transgenic studies have implicated various 3' untranslated nucleotide sequence regions in the OT and AVP genes which appear to contain these regulatory elements. A structural analysis of these regions in the mouse genes will be discussed. A second problem has been the availability of appropriate model systems to test constructs for "cell-specific" expression. Transgenic mice have been effective models in this regard but are costly and labor-intensive. An alternative approach using biolistic techniques to transfect slice-explants containing the primary OT and AVP neurons in culture will be described. PMID- 8713948 TI - Functional domains in the oxytocin gene for regulation of expression and biosynthesis of gene products. AB - In the oxytocin (OT) gene several regions can be discerned that have a function in regulating its expression. Firstly, in the proximal 5' flanking region regulatory elements have been discovered that are targets for transcription factors of the nuclear hormone receptor family. Through these elements the OT gene of rat and man is responsive to estrogens, thyroid hormones and retinoids. Furthermore, these elements can be employed by the nuclear hormone orphan receptor family for repressive or inductive actions. In the distal 5' flanking region the POU class III proteins Brn-1, Brn-2, Brn-4, that are expressed in magnocellular neurons, and Oct-6 are able to bind, but do not display a significant regulatory activity on the OT gene in heterologous expression systems. Secondly, the OT precursor harbours both the biologically active hormone and the protein neurophysin that is able to associate with the hormone. Heterologous expression of wild-type and mutant vasopressin cDNAs in peptidergic cell lines shows that the highly homologous vasopressin-associated neurophysin domain associates with the hormone domain within the prohormone. This intramolecular interaction between two prohormone domains serves an essential intracellular function, i.e. the proper sorting of the prohormone into the regulated secretory pathway. PMID- 8713950 TI - Stimulus-specific expression of inducible transcription factors in identified oxytocin neurones. AB - The activation of the magnocellular oxytocin system by different physiological stimuli will require specific genomic responses that may or may not reflect the electrical and short-term secretory activity of the neurones. One of the main determinants of synthetic activity is the rate of transcription and this can be altered acutely by the action of inducible transcription factors (iTFs). Having shown that the expression of two iTFs, the protein products of the c-fos and c jun genes, does not correlate directly to the electrical activity of magnocellular neurones (Luckman et al., 1994) the expression of leucine zipper iTF mRNAs was measured following different stimuli using combined radioactive and non-radioactive in situ hybridization. Stimuli that are dependent on brainstem afferents such as parturition and systemic injection of cholecystokinin caused co induction of c-fos and c-jun in oxytocin neurones. Mild osmotic stimulation, a stimulus dependent on forebrain afferents, induced c-fos, fos B and jun B, but inhibited c-jun. Similar patterns of leucine zipper iTF expression have been noted in cultured cells following activation of protein kinases C and A, respectively. Input from the brainstem appears to be mediated, at least in part, by noradrenaline acting on alpha(1)-adrenoceptors. While the forebrain inputs are not well characterised they do appear to include a glutaminergic component that may activate a variety of receptors. Interestingly, another member of the leucine zipper family known to be induced by protein kinase A, inducible cAMP early repressor (ICER), that was previously thought to be restricted to the pineal gland, was expressed in magnocellular neurones following osmotic stimulation but not parturition. Furthermore, the differential expression of iTFs is not limited to this family. Osmotic stimulation influences c-fos, but it also causes the expression of NGFI-A and NGFI-B, members of the zinc finger family of iTFs. By contrast, an acute suckling stimulus is able to induce c-fos and NGFI-A, but not nGFI-B. PMID- 8713949 TI - Effect of gonadal steroids upon hypothalamic oxytocin expression. AB - Oxytocin (OT) has been implicated in reproductive events. Gonadal steroids, which are also essential for reproduction, may in these instances influence the expression of the OT gene. The relationship between gonadal steroids and OT expression is incompletely understood. First, although estrogen (E) induces OT expression in heterologous in vitro systems and the OT gene contains an estrogen response element, evidence for a stimulatory effect of E upon OT mRNA in vivo as in the rat hypothalamus has been lacking. Second, an enhanced and coordinated expression of OT and vasopressin (AVP), mRNA during late pregnancy and lactation is not expected. This is especially true in lactation, during which suckling selectively activates OT, not AVP, neurons. We now report that E does increase OT mRNA levels in the rat hypothalamus. Ovariectomized rats were implanted initially with E, followed by progesterone (P) 48 hrs later; both were sustained for 12 days and the P implants were removed 48 hrs prior to sacrifice. By in situ and Northern blot hybridizations, OT, but not AVP, mRNA was increased in the hypothalamus of animals receiving this regimen compared to sham treated cohorts. A similar phenomenon associated with exposure to E and P followed by P withdrawal occurs in physiologic states such as day 21 of pregnancy, day 10-12 of lactation, and with interruption of the suckling stimulus for 48 hrs. If E was not administered or if P was not withdrawn prior to sacrifice, no increase was found in OT mRNA. Our data highlight the pivotal role of P withdrawal in the induction of the hypothalamic OT gene in the E-primed rat. PMID- 8713951 TI - Production of hybrid oxytocin/vasopressin precursors and accumulation of oxytocin precursors in the rough endoplasmic reticulum of rat magnocellular neurons. AB - Most magnocellular hypothalamic neurons synthesize the precursor for either vasopressin (AVP) or oxytocin (OT). The AVP precursor is cleaved to give AVP, AVP associated neurophysin (AVP-NP) and a glycopeptide (GP), whereas the OT precursor gives OT and OT-NP. In Brattleboro rats a frame-shift mutation in the AVP-NP encoding region of the gene prevents the secretion of AVP by the cells and, in most AVP neurons, AVP itself is virtually undetectable. A small number of magnocellular neurons in homozygous Brattleboro rats contain very large accumulations of peptide in distended saccules of rough endoplasmic reticulum (RER), and this peptide is immunoreactive for AVP and C-terminal OT-NP, but not for OT, AVP-NP or GP (Pow et al., 1992). We have now shown that this results from somatic non-homologous crossing over of the AVP and OT genes, resulting in the production of hybrid mRNA molecules with the 5'end of the AVP sequence and the 3' end of the OT sequence (AVP/OT transcripts). In most cases, the crossing over occurs within the highly homologous B exons (Mohr et al., 1994). In addition to the production of AVP/OT hybrid transcripts, polymerase chain reaction (PCR) amplification of mRNA from the hypothalami of homozygous rats also reveals OT/AVP hybrid transcripts, with 5' OT sequences and 3' AVP sequences. Furthermore, both types of hybrid transcript are not restricted to homozygous Brattleboro rats but can also be found in normal Long Evans animals. To date, we have not been able to locate cells in which the OT/AVP hybrids are produced; all the magnocellular neurons with hybrid peptide accumulations in the RER so far studied have been shown by immunocytochemistry to be of the AVP/OT type. In both normal and homozygous Brattleboro rats large accumulations of peptide do occur in the RER of OT-producing neurons but the peptide is immunoreactive for OT and OT-NP but not for AVP, AVP-NP or GP. Such cells increase in number 10-fold after injection of 20 micrograms estradiol daily for 7 days (Pow et al., 1991). Why this apparently normal gene product accumulates within the RER remains to be determined. PMID- 8713952 TI - Sense- and antisense-targeting of oxytocinergic systems in the rat hypothalamus. AB - 3' phosphorothioate modified sense or antisense oligonucleotides to oxytocin transcripts were used for in vivo targeting of oxytocin (OT) neurons in the rat hypothalamus. Intracerebroventricular injections of antisense probe resulted in a loss of systemic OT. However, abundant immunoreactive OT as well as oxytocin mRNA hybridization signal was visualized in the hypothalamo neurohypophysial system (HNS) of these animals. RT-PCR of hypothalamic homogenates revealed clearly detectable amounts of cytoplasmic OT mRNA in spite of sense or antisense treatment. Immunostaining with an antibody to DNA-RNA triple helix resulted in cytoplasmic reaction product in the HNS in the antisense group, which was not found when tissue sections had been pretreated with RNase. Animals injected with the sense probe showed a less pronounced but significant loss of systemic OT while immunoreactivity for this peptide in the posterior lobe seemed to be unaffected. RT-PCR of OT encoding mRNA extracted from sense injected rats indicated that these transcripts were of smaller size than samples from antisense treated animals or controls. Immunostaining with the triple helix antibody revealed distinct immunoreactive dots in cellular nuclei throughout the brain in the sense group. Our findings suggest that sense and antisense probes may not readily be employed as "functional antagonists" since peptidergic neurons are probably capable of responding in various ways to the treatment. RNase H may be less important in hypothalamic neurons as commonly suggested. Targeted transcripts are likely to form complexes which may somehow interact with secretion. Triple helix formation in the nucleus may not be able to induce an efficient transcriptional arrest. Although endocrine and behavioral changes observed in antisense treated animals seem to confirm the hypothesis that a selective translational "knock out" can be achieved with in vivo hybridization strategies, the actual underlying molecular events are far from being understood. On the other hand, sense or antisense strategies may provide valuable insights into molecular and cellular events associated with neurosecretion. PMID- 8713953 TI - Oxytocin transgenic mice. AB - We have compared the expression patterns in transgenic mice of bovine oxytocin constructs consisting of the 0.9 kilobase pair (kbp) structural gene flanked by varying lengths of upstream and downstream sequences. Over 200 offspring were derived from fertilized one-cell mouse eggs injected with construct bOT6.5, which consists of 3 kbp of upstream sequences and 2.6 kbp of downstream sequences. However, no transgenic founders were identified. In parallel experiments with other constructs, 30% of pups carried integrated copies of the injected transgene DNA. It therefore appears that bOT6.5 is toxic to mouse embryos. As previously reported (Ang et al., 1991), bOT, consisting of 2.6 kbp of downstream sequences and 0.6 kbp of upstream sequences, was expressed in lung and in testicular Sertoli cells, but no expression was detected in the hypothalamus. bOT3.5, which consists of 0.6 kbp of upstream sequences and 1.9 kbp of downstream sequences, retains testis and lung expression, but, surprisingly, is also expressed in the hypothalamus. These data suggest that the 0.7 kbp of downstream sequences that are present in bOT, but which are absent from bOT3.5, contain elements that mediate the repression of hypothalamic expression. The activity of this repressor must be itself overcome in the normal genomic context of the bovine OT gene. Within the hypothalamus, in situ hybridisation analysis has revealed expression of bOT3.5 in the supraoptic nucleus (SON) and paraventricular nucleus (PVN), but not in the suprachiasmatic nucleus (SCN). In parallel with the response of the endogenous murine OT RNA, 7 days of salt-loading resulted in a significant increase in the level of transgene RNA in the SON. In the PVN, neither the endogenous OT RNA nor the transgene RNA responded significantly to salt-loading. Transgene RNA levels in the hypothalamus have also been shown to be elevated during late pregnancy and lactation. PMID- 8713954 TI - Regulation of oxytocin expression in the bovine corpus luteum. Orphan receptors and the oxytocin promoter. AB - The bovine oxytocin gene is massively up-regulated during the early development of the corpus luteum. Oxytocin transcription is induced in a highly synchronous fashion in the granulosa cells of the dominant follicle at the time of ovulation. The possibility to isolate large numbers of differentiating granulosa-luteal cells from exactly defined stages of development allows the investigation of the factors controlling oxytocin expression in vivo by molecular and cell biology methods. Using primary cultures of bovine granulosa cells the synergistic activation of oxytocin transcription by the cAMP pathway and stimulation of IGF-I or insulin receptors could be established. Analysis of transcription factors isolated from the nuclei of bovine granulosa cells and corpus luteum led to the identification of the tissue-specific orphan receptor SF-1 binding to the promoter of the actively transcribed oxytocin gene. The luteinizing bovine granulosa cells provide the only easily accessible experimental system established so far in which the endogenous oxytocin gene is expressed. Although the link between increased cAMP level and receptor tyrosine kinase activation on the one hand and the induction of oxytocin transcription on the other has not been established yet, these experiments constitute one of the few direct approaches to investigate the complexity of events that regulate oxytocin expression in vivo. PMID- 8713955 TI - Oxytocin mRNA in the human paraventricular nucleus in AIDS. PMID- 8713956 TI - Daily rhythms of oxytocin secretion. PMID- 8713957 TI - Opioid-noradrenergic interactions in the control of oxytocin cells. AB - The nucleus tractus solitarii (NTS) projects directly to the oxytocin neurones of the supraoptic nucleus (SON), and relays afferent stimuli arising from the birth canal during parturition. About 80% of these projecting neurones are noradrenergic, and these same neurones are activated following systemic administration of cholecystokinin (CCK), which also results in an increased electrical and secretory activity in oxytocin neurones. Oxytocin release in response to CCK is abolished following selective neurotoxic destruction of these noradrenergic neurones. Oxytocin release following CCK (and that during parturition) is potently inhibited by morphine, which blocks the local noradrenaline release in the supraoptic nucleus. This acute opiate action involves presynaptic inhibition of the noradrenergic terminals, and occurs without marked suppression of the activity of noradrenergic cells in the NTS. During chronic exposure to morphine the oxytocin system becomes tolerant to, and dependent upon morphine. In the course of tolerance, oxytocin cell activation in response to CCK recovers from initial inhibition. However, the pathway that mediates this response does not appear to become dependent: the oxytocin cell response to CCK is unchanged by opiate withdrawal induced by naloxone, despite a large increase in the background electrical activity of oxytocin cells provoked by withdrawal. Nevertheless, expression of withdrawal excitation by oxytocin neurones is curiously contingent upon the activity of the noradrenergic input in that prior lesioning of this input has no effect upon the subsequent withdrawal excitation of oxytocin cells. Yet under urethane anaesthesia, acute pharmacological blockade of the noradrenergic input suppresses withdrawal. We discuss how these paradoxical observations might be reconciled, and note that the difference may be related to differing levels of tonic activity in the noradrenergic input. It is possible that dependence relies upon the input when it is there, but not when it is not. PMID- 8713958 TI - Oxytocin suppresses the GABAergic synaptic input in supraoptic neurones from the rat. AB - Oxytocin, acting as an autotransmitter, gives rise to release of calcium from intracellular store(s) within magnocellular neurons in the supraoptic nucleus (SON). A possible target for a rise in intracellular calcium is the GABAA receptor, since it is known that the functioning of this receptor may depend (directly) on the intracellular free calcium concentration. Therefore the effect of oxytocin on the GABAergic synaptic input in the SON was analyzed. In situ patch clamp recordings from individual neurons of the SON were performed. Spontaneously occurring, bicuculline sensitive GABAergic inhibitory synaptic currents (IPSCs) were pharmacologically isolated from the excitatory glutamatergic synaptic input. This isolated GABAergic synaptic input was spontaneously and tonically active, arising from both somatic as well as from dendritic synaptic contacts and operated a chloride conductance. Application of oxytocin during such recordings, strongly reduced the amplitude of the IPSCs in 73% of the recordings. This reduction was (i) completely reversed by washing, (ii) blocked by a specific oxytocin receptor antagonist, and (iii) observed in slices from both female and from male animals. Thus autotransmission involving disinhibition of magnocellular neurons may explain why oxytocin facilitates it own release. PMID- 8713959 TI - Influence of reproductive state and ovarian steroids on facilitation of the milk ejection reflex by central oxytocin. AB - In the rat, the synchronous bursting activity of oxytocin neurones associated with the milk-ejection reflex displays important changes during the peri-partum and lactational periods. The most dramatic of these changes is the appearance of a facilitatory response to centrally-administered oxytocin, involving an increase in the frequency and amplitude of bursting in the oxytocin neurones, as well as elevation of their background activity. Studies of rats at different times in the pre- and post-partum period show that this response first appears on day 3 of lactation. Ovariectomy on day 21 of gestation, or treatment with the anti oestrogen tamoxifen on day 22, does not prevent the appearance of this response. However, ovariectomy and treatment with ovarian steroids for 3 days prior to parturition can dramatically alter the character of the facilitatory response. Oestradiol treatment causes an early (pre-partum) appearance of the facilitatory response, whereas progesterone causes the appearance of an inhibitory response (reduction in milk-ejection frequency) to central oxytocin. A major target for the central effects of oxytocin are the bed nuclei of the stria terminalis (BST) and modulation of the neuronal responses in this region may, in part, underlie the changing facilitatory effects. In vitro recordings indicate that sensitivity of BST neurones to oxytocin is increased between pregnancy and lactation, and oestradiol treatment enhances responsiveness coincident with the appearance of a facilitatory response. Progesterone pre-treatment also increases the ability of BST neurones to respond to oxytocin in vitro (although less than oestradiol), an unexpected result given the absence of oxytocin-induced facilitation of the milk ejection reflex in late pregnancy or following progesterone treatment in vivo. In vivo recordings of BST neurones suggest that one explanation of this lack of correlation may reside in the presence of a mechanism which attenuates the excitatory response to oxytocin, perhaps serving to prevent premature expression of the facilitatory action of oxytocin. Collectively, these data show that there are dramatic reproductive state and steroid-dependent changes in the central action of oxytocin on the synchronous bursting of magnocellular oxytocin neurones. These changes, which have important consequences for the optimization of bursting in oxytocin neurones, may involve plasticity of transduction mechanisms in the oxytocin-responsive elements of the limbic system. PMID- 8713961 TI - Physiologically-linked structural plasticity of inhibitory and excitatory synaptic inputs to oxytocin neurons. AB - The adult oxytocinergic system undergoes extensive synaptic and neuronal-glial remodelling in response to differing conditions of secretion and has become a remarkable example of activity-dependent structural plasticity in the adult mammalian brain. Under stimulation (parturition, lactation, chronic dehydration), glial coverage of oxytocin neurons is significantly reduced and their surfaces become extensively juxtaposed; concurrently, they are contacted by an increased number of synapses. These changes are reversible with cessation of stimulation. We here present observations showing that putative inhibitory and excitatory afferents contribute to this synaptic plasticity. The data are derived from several different comparative analyses of ultrathin sections of the rat supraoptic nucleus (SON) in which presynaptic (gamma-amino butyric acid (GABA) or glutamate) and postsynaptic (oxytocin or vasopressin) partners were identified with postembedding immunogold staining. We thus found that in virgin rats, under basal conditions of oxytocin release, 30-40% of synapses on oxytocinergic or vasopressinergic somata in the SON are GABAergic and about 20% glutamatergic. On the other hand, in lactating rats, in which oxytocin secretion is greatly enhanced, there was an increase in the incidence of both types of synapses, and in particular, on those impinging on oxytocinergic somata. PMID- 8713960 TI - The central oxytocin pulse generator: a pacemaker for luteolysis. AB - Oxytocin (OT) is released from the neurohypophysis into the jugular vein of sheep in small 1-2 min pulses (ca. 10 pg/ml) in both cyclic and ovariectomized sheep. In intact cycling sheep, additional hour long bursts of OT (up to 200 pg/ml) occur in peripheral blood during luteolysis at intervals of 6 to 9 hrs which appear to regulate large luteolytic pulses of uterine prostaglandin F2a (PGF2a). Since the ovine corpus luteum (CL) also synthesizes OT, experiments were performed to distinguish between the relative contributions of the neurohypophysis and the CL to the large bursts of OT secreted during luteolysis. Two models were used. First, ovariectomized sheep were given exogenous E and/or P by constant infusion to simulate levels during the estrous cycle. Second, in tact cycling sheep, the CL was surgically excised during the luteal phase to exclude the CL as a source of OT and, at the same time, subject the animals to the withdrawal of P. Pulses of OT in jugular vein plasma were determined by RIA or biometry of the uterus. The findings are summarized as follows: In ovariectomized sheep, maintained on low E (0.05 g/hr) to preserve the OT pulse generator, infusion of E (1 microgram, 2 micrograms or 4 micrograms/hr) for 12 to 36 hr, caused a series (4 to 6) of rapid increases in OT pulse frequency each lasting 1 to 2 hrs at intervals of 3 hrs. The time of onset of high frequency pulses was dose-dependent. Withdrawal of 10 day infusions of P (500 micrograms/hr) superimposed on low E (0.05 microgram/hr) also evoked a series of high frequency episodes of OT pulses beginning 24 hrs after P withdrawal. In intact sheep, surgical removal of the CL resulted in a series of high frequency pulses similar in duration and frequency to those following the withdrawal of P in the ovariectomized animal. We conclude that: (1) an increase in E or returning E action causes the OT pulse generator to alter its frequency intermittently thus producing a series of 4 to 6 episodes of high frequency pulses of OT. (2) Similar changes can be evoked by withdrawal of P either by terminating an infusion of P in the presence of E in the ovariectomized sheep or by surgically removing the CL from the ovary in the intact sheep. (3) At the end of the reproductive cycle, the central OT pulse generator appears to act as a pacemaker which, acting on the endometrial OT receptors, triggers a series of pulses of PGF2a from the uterus and hence causes regression of the CL. In the sheep and other ruminants, an intermittent supplemental secretion of OT from the CL, triggered via the central OT pulse generator, may also be required to amplify the luteolytic pulses of PGF2a from the uterus. (4) In addition to the well established interaction of ovarian steroid hormones, and the hypothalamic pituitary system for the initiation of the reproductive cycle via the gonadotrophins, there is now good evidence for an interaction of ovarian steroids and the posterior pituitary for terminating the reproductive cycle. PMID- 8713962 TI - Release of oxytocin within the supraoptic nucleus. Mechanisms, physiological significance and antisense targeting. AB - In addition to its secretion from neurohypophysial terminals, oxytocin (OT) is released within the hypothalamic magnocellular nuclei--the supraoptic (SON) and paraventricular nuclei--in response to various stimuli as mainly shown by microdialysis studies. During parturition and suckling, OT is released within the SON in a positive feedback role to further amplify its own local and/or neurohypophysial release depending upon the reproductive conditions. Whereas, for instance, intra-SON OT is important for appropriate OT secretion and milk ejection during suckling, it seems to be involved in the timing of the parturition process without directly affecting OT secretion during birth. As a consequence of various morphological and physiological adaptations of the OT system during lactation, the release pattern of OT into blood and within the SON is altered in response to other stimuli, like swim stress and peripheral osmotic stimulation. In addition, in lactating, but not virgin female or male rats, unilateral stimulation of the SON results in the release of OT, but not vasopressin or various amino acids, within the contralateral SON, which implicates a functional role of intranuclear OT in the plastic coupling of the paired SON during lactation. To manipulate OT synthesis and release, antisense targeting was used. Novel, acute effects (within 5 hours) of an OT antisense oligodeoxynucleotide on suckling-related parameters and, specifically, on the electrophysiological excitability of OT neurons as well as their ability to express the Fos-protein after CCK-8 are described. Since the OT content in the hypothalamo-neurohypophysial system was still unaffected at this time, antisense oligonucleotides may induce feedforward alterations in neuronal responsiveness which precede or parallel effects on neuropeptide synthesis. Taken together, the functional significance of OT released within the brain and into blood during reproduction provides an example of how a neuropeptide might regulate complex physiological and behavioral performances in a synergistic manner. PMID- 8713963 TI - Somatodendritic autoreceptors on oxytocin neurones. AB - Oxytocin (OT) is known to exert a facilitatory effect on its own pulsatile release during the milk ejection reflex. Several experimental approaches strongly suggested that this effect occurred in the supraoptic and paraventricular magnocellular hypothalamic nuclei where the OT neurones are concentrated. In these nuclei autoradiographic techniques failed however to detect OT receptors. In the presently reported data, OT binding sites were visualized in the supraoptic, paraventricular and accessory magnocellular nuclei in lactating rats, suckled or not, after acute intracerebroventricular injection of an OT antagonist. No such labelling occurred either in untreated lactating rats or in OT antagonist treated male rats. The combined application of histoautoradiographic and immunocytochemical techniques revealed that the OT binding sites were concentrated on magnocellular OT cell bodies and dendrites but not on their axons and endings in the pituitary neural lobe. The presently detected somatodendritic autoreceptors on OT neurones probably mediate the facilitatory effect of OT on its own release during the milk ejection reflex. PMID- 8713964 TI - Progesterone suppression of activation of supraoptic nucleus (SON) neurones in late pregnancy in the rat. PMID- 8713965 TI - Concurrent changes in the response of limbic neurones and mammary gland to exogenous oxytocin in late pregnancy. PMID- 8713966 TI - Appearance of the facilitatory action of central oxytocin on milk-ejection. Independence from peri-partum steroidal changes. PMID- 8713967 TI - Importance of noradrenergic input in morphine withdrawal excitation of supraoptic oxytocin neurones in urethane-anaesthetised rats. PMID- 8713968 TI - Rapid stimulation of oxytocin gene expression in supraoptic nucleus neurones during morphine withdrawal in rats. PMID- 8713969 TI - The excitatory effect of systemic cholecystokinin on oxytocin neurones is unchanged by morphine dependence and withdrawal. PMID- 8713970 TI - Establishing behavioral and physiological functions of central oxytocin: insights from studies of oxytocin and ingestive behaviors. AB - Plasma oxytocin (OT) levels are strongly correlated with inhibition of ingestion in many models of stimulated food and NaCl intake in rats, but peripheral administration of OT or OT antagonists has little or no effect on these behaviors. These findings led us to propose that central OT secretion from parvocellular neurons occurring in parallel with pituitary secretion from magnocellular neurons acts to inhibit ingestion of both food and salt. Multiple lines of evidence now support this hypothesis: 1) intracerebroventricular (icv) OT administration inhibits food intake in fasted rats and NaCl intake in hypovolemic rats; 2) icv administration of OT-receptor antagonists significantly blunts the effects of anorexigenic agents on food intake and the action of naloxone to inhibit hypovolemia-induced intake of NaCl, but not water; 3) most treatments that inhibit food and/or NaCl intake stimulate expression of c-fos in parvocellular as well as magnocellular OT neurons, indicating simultaneous activation of both centrally-projecting and pituitary-projecting OT neurons; 4) icv treatment with cytotoxic conjugates of ricin A and OT to disable cells bearing OT receptors leads to a disinhibition of NaCl intake similar to that produced by OT antagonists; 5) administration of ethanol, a well known inhibitor of OT secretion, produces effects on stimulated food and NaCl intake in rats analogous to those produced by OT-antagonists and ricin-OT conjugates. In conjunction with studies demonstrating natriuretic effects of circulating OT, these results therefore support the concept of coordinated central and peripheral OT secretion as a mechanism for regulating body solute homeostasis in rats. These phenomena will be used as a framework to discuss and critically evaluate the criteria that are both necessary and sufficient to firmly establish behavioral and physiological functions of centrally-secreted peptides such as OT. PMID- 8713971 TI - Oxytocin and the molecular basis of monogamy. AB - Previous studies in rats have implicated central oxytocin (OT) pathways in the onset of maternal behavior, female sexual receptivity, and the response of the pups to social separation. However, the rat is not ideal for studying effects of OT on attachment as rats fail to form selective, enduring social bonds. To study male-female pair bonds, our laboratory has focused on a microtine rodent, the prairie vole, which is monogamous and highly affiliative. Adult prairie voles form pair bonds after mating (with prolonged, repeated bouts of copulation). As mating releases OT in several species of mammals, we hypothesized that this release was important for pair bond formation in the prairie vole. Central administration of an OT antagonist (but not a V1 antagonist) prevents pair bonding without interfering with the mating behavior. Moreover, central infusion of OT (but not vasopressin, AVP) facilitates pair bonding n the absence of mating. In males, it is AVP (not OT) that appears necessary for pair bond formation. The pattern of OT (and AVP) receptor distribution in the prairie vole brain is entirely distinct from the pattern observed in the closely related non monogamous montane vole. OT receptors (OTR) in these two species show virtually identical kinetics, specificities, and cDNA sequences (RNA from parturient uterus). In current studies, we are screening genomic libraries from prairie and montane voles to determine if species differences in OTR promoters account for the strikingly different patterns of regional expression in brain. These studies should ultimately provide insight into a neuroendocrine mechanism for pair bond formation. PMID- 8713972 TI - Estrogen modulation of oxytocin and its relation to behavior. AB - Establishment of the behavioral significance of oxytocin neurotransmission in the brain has been a leading component in the emerging concept of neuropeptide regulation of behavior. Elucidating the behavioral effects of oxytocin has been facilitated by its profound regulation by estrogen in discrete brain areas and its subsequent role in estrogen-dependent responses. For example, female sexual behavior is estrogen dependent, estrogen markedly increases oxytocin binding in the ventromedial nucleus of the hypothalamus and infusion of oxytocin in this brain area increases female sexual behavior. A similar correlation exists for the role of oxytocin and estrogen in the regulation of maternal behavior. A possible underlying cause of these behavioral effects is that by acting as an anxiolytic, oxytocin reduces the inhibition inherent in social encounters. Behavioral tests in the laboratory frequently involve the exposure of the animal to a novel environment, such as a pup-retrieval apparatus or mating arena, combined with exposure to an unfamiliar conspecific. These stimuli are likely to induce a stress response and perhaps this anxiety is reduced by oxytocin. Recent evidence in mice suggests that oxytocin has anxiolytic properties in estrogen-treated females (McCarthy and Goldman, 1994) and supports the hypothesis that a unifying principal in oxytocin action in the brain is to facilitate social encounters by reducing the associated anxiety. PMID- 8713973 TI - Oxytocin-induced penile erection. Role of nitric oxide. AB - In order to investigate the mechanism of action by which oxytocin induces penile erection, the effect of NG-nitro-L-arginine methyl ester (NAME) and NG-monomethyl L-arginine (NMMA), inhibitors of nitric oxide (NO) synthase, injected into the paraventricular nucleus of the hypothalamus (PVN) on the response to oxytocin injected into the PVN was studied in male rats. NAME and NMMA, but not NG-mono methyl-D-arginine (D-NMMA), which does not inhibit NO-synthase, prevented in a dose-dependent manner the response to oxytocin. NAME was 4-5 times more potent than NMMA. NAME prevention of the oxytocin effect was not observed when NAME was given together with L-arginine but not with D-arginine. Oxytocin-induced penile erection was prevented by the oxytocin antagonist d(CH2)5Tyr(Me)-Orn8-vasotocin and by methylene blue, an inhibitor of guanylate cyclase, but not reduced hemoglobin, a NO scavenger, given intracerebroventricularly (i.c.v.). In contrast, both methylene blue and hemoglobin were ineffective when injected into the PVN, unlike d(CH2)5Tyr(Me)-Orn8-vasotocin. Penile erection was induced also by sodium nitroprusside and hydroxylamine, two NO donors, injected into the PVN. Like the oxytocin effect, the NO donor response was prevented by i.c.v. d(CH2)5Tyr(Me)-Orn8-vasotocin and methylene blue, but not hemoglobin. In contrast, the three compounds were ineffective in preventing the NO donor response when injected into the PVN. The present results suggest that oxytocin induces penile erection by activating NO synthase in the PVN. NO in turn activates oxytocinergic neurons projecting to extra-hypothalamic areas that control the expression of this male sexual function by a guanosine cyclic 3':5' monophosphate (cGMP) independent mechanism at least in the PVN. PMID- 8713974 TI - Guinea pigs (Cavia apera f. porcellus): do they show pair-bonding behavior in relationship to oxytocin? PMID- 8713975 TI - Reduction of plasma oxytocin levels in patients suffering from major depression. PMID- 8713976 TI - Investigation of the oxytocin receptor at the molecular level. AB - Before the onset of labor, the myometrium becomes extremely sensitive to oxytocin, for which it is a primary target tissue, because of a tremendous increase in the number of oxytocin receptors. To study this dynamic expression and regulation of the receptor, we cloned the human oxytocin receptor cDNA in 1992 and characterized its expression in female reproductive organs and in the rat brain using the receptor cDNA and anti-oxytocin receptor monoclonal antibodies. In the uterus, we revealed that oxytocin receptor expression is mainly regulated at the transcriptional level. For the investigation of this transcriptional regulation, we isolated and characterized the human oxytocin receptor gene. The gene contains 3 introns and 4 exons and is assigned to 3p26.2 of human chromosome 3. In the 5'-upstream region of this gene, we found several known binding sites for transcription regulation factors including the nucleofactor-interleukin 6 binding consensus sequence and an acute phase reactant responsive element. Our findings on the structure of the oxytocin receptor gene should help to elucidate the mechanism by which expression of the gene is induced dramatically at parturition in the uterus and how the gene is regulated in other organs such as the mammary gland and central nervous system. PMID- 8713977 TI - CNS oxytocin receptor mRNA expression and regulation by gonadal steroids. AB - A genomic clone was used to generate a rat oxytocin receptor (OTR) probe for in situ hybridization studies in order to monitor changes in expression of OTR mRNA in the CNS of male and female rats in brain regions reported to be rich in OTR binding. This probe predominantly detects expression in the ventromedial hypothalamus (VMH). Quantitatively greater expression of OTR mRNA was more evident in the male VMH than in that of the female. OTR mRNA expression in the VMH is also enhanced in gonadectomized rats treated with either estrogen or testosterone. Previous studies of VMH OTR mRNA expression during the estrous cycle revealed a role of estrogen in proestrous OTR mRNA expression and also provided evidence for progesterone's modulatory effect on OTR expression. A related study which examined OTR mRNA expression during gestation, parturition, and lactation further supported the regulation of the OTR by gonadal steroids, as a pronounced induction of expression occurred at parturition when estrogen levels were highest. These results support previous studies which have documented the regulation of OTR binding by gonadal steroids and suggest that this regulation may be the result of altered expression of the OTR gene. Induction of OTRs in anatomically distinct regions of the VMH may be important in promoting OT'ergic neurotransmission required for sexual behavior or the induction of gonadotropin release by OT, which requires estrogen and progesterone priming. The anatomical variation in OTR mRNA localization and its restricted detection in the VMH by this probe suggests potential CNS OTR heterogeneity. PMID- 8713979 TI - Structure and organization of the bovine oxytocin receptor gene. AB - A DNA probe specific for the V and VI transmembrane domains of the bovine oxytocin receptor was initially prepared by reverse transcription PCR, and its structure and specificity confirmed by DNA sequencing. This probe was then used to screen a bovine genomic DNA library in bacteriophage lambda, and three positive clones were purified, subjected to restriction analysis and relevant fragments sequenced. Parallel to this, a cDNA library prepared using bovine endometrial RNA at the time of ovulation was screened by PCR employing the same primers as above. The longest cDNA clone was also fully sequenced. This clone still lacked, however, a substantial stretch of 5'sequence. The full transcript structure, and hence also the exon-intron organization, was then obtained by RT PCR using primer oligonucleotides deduced from the cloned genomic sequence. All nucleotide sequence information was derived from a combination of two independent genomic clones, a cDNA clone and several independent RT-PCR reactions programmed by myometrial RNA, all in both strand orientations. The structural organization of the bovine oxytocin gene essentially conforms to that described for the human gene. Unlike the human gene, however, the 5'non-coding region of the primary transcript is interrupted by only a single intron, with a further intron in the coding region separating the sequences encoding the transmembrane domains VI and VII. The difference between this structure and that for the human gene suggests the existence of a differential splicing of 5' non-coding sequences. PMID- 8713978 TI - The sheep endometrial oxytocin receptor. AB - The sheep endometrial oxytocin receptor plays a central role in determining the time at which luteolysis occurs during the oestrous cycle, and in the events leading to the establishment of pregnancy (the maternal recognition of pregnancy). Expression of the receptor in the uterus is controlled by ovarian steroid hormones, and by trophoblast interferon (IFN-tau). We report here studies on the second messengers involved in the effect of IFN-tau, and on the structure and expression of the oxytocin receptor. The receptor is expressed in ovine endometrial explants during culture, when the explants are taken during the luteal phase of the cycle; this process is partially blocked by inhibitors of protein kinase C, or by down-regulation of protein kinase C. Therefore it is suggested that protein kinase C, rather than tyrosine kinases, is involved in the effect of IFN-tau on oxytocin receptor expression. Northern blotting shows that in common with uterine oxytocin receptor mRNA in other species, the message is heterogeneous. cDNA sequencing indicates the sheep uterine oxytocin receptor is at least 2 amino acids longer than those of other species, and expression of the receptor in Cos-7 cells induces oxytocin responsiveness in terms of phosphoinositide turnover. PMID- 8713980 TI - Identification of agonist binding sites of vasopressin and oxytocin receptors. AB - The present study aims at delineating residues in the vasopressin/oxytocin receptor family responsible for the high affinity binding of the hormone. Therefore, we have constructed a computer-generated 3 dimensional model of the rat V1a vasopressin receptor subtype which allowed us to propose residues likely to be involved in agonist binding. Among these residues, several are highly conserved in the receptor family. They were selected for site-directed mutagenesis on the basis of putative direct interaction with bound ligands. The present model and experimental results led us to conclude that the hormone is docked in a pocket completely buried in the transmembrane core of the receptor. Large polar residues, such as glutamine and lysine, located in transmembrane regions 2,3,4 and 6 are involved in the binding of the neurohypophysial hormone. Since all the mutated residues are highly conserved in AVP and OT receptors, we propose that the agonist binding site is similar in all members of the receptor family; only minor changes were found in antagonist potencies, suggesting that agonist and antagonist binding sites do not completely overlap. PMID- 8713981 TI - Conversion of the myometrial oxytocin receptor from low to high affinity state by cholesterol. AB - Reconstitution experiments with the myometrial oxytocin receptor from guinea pig showed a strong requirement of the oxytocin receptor binding function for the presence of cholesterol in preformed liposomes. To investigate the effect of cholesterol on the oxytocin receptor in the plasma membrane, a new method to modify the membrane cholesterol content was developed. With substituted cyclodextrins we were able to selectively deplete the myometrial plasma membrane of cholesterol. Vice versa, incubation of cholesterol-depleted membranes with a soluble cholesterol cyclodextrin complex restored the cholesterol content of the plasma membrane. Binding experiments showed, that with the removal of cholesterol from the membrane, the oxytocin receptor was converted from high to low affinity state. Increasing the cholesterol content of the membrane again restored the high binding affinity. Substitution of membrane cholesterol with other steroids showed a strong dependence of the oxytocin receptor function on the structure of the cholesterol molecule. Experiments with the detergent solubilized oxytocin receptor and with oxytocin receptors expressed in the Baculovirus/Sf9 cell system provided further evidence for a direct interaction of the oxytocin receptor with cholesterol. PMID- 8713982 TI - Molecular basis for agonist selectivity in the vasopressin/oxytocin receptor family. AB - Vasopressin (AVP) and oxytocin (OT) are two nonapeptides differing at position 3, in the cyclic part of the peptide, and at position 8, in the C-terminal tripeptide. In this study, we have evaluated the interactions between these two positions of the hormones and the oxytocin receptor (OTR), the V1a and the V2 vasopressin receptors. The contribution of these two positions to receptor selectivity was analyzed by using several peptide analogues bearing substitutions at either position 3 or 8. The putative interactions between receptor residues and hormone residues at position 3 and 8 were then deduced by using a three dimensional model of the neurohypophysial hormones docked into their respective receptors. On the basis of this model, we found that the lateral chain of residue 8 might interact with residues located in the first extracellular loop. By using site-directed mutagenesis on the cloned receptors, we identified a non-conserved residue in the first extracellular loop that interacts with the lateral chain of residue 8 in the hormone. We demonstrated that this interaction is crucial for receptor selectivity to the different agonists. PMID- 8713983 TI - Oxytocin receptor gene expression in female rat kidney. The effect of estrogen. AB - Using in situ hybridization methods that discriminate mRNAs encoding rat vasopressin V1a, V1b, V2 and oxytocin receptors in hepatic, brain and renal tissues, experiments were done to determine whether estrogen and/or progesterone influence renal vasopressin receptor (VR) or oxytocin receptor (OTR) transcripts. Estrogen induced OTR gene expression in the outer stripe of the outer medulla and increased expression of OTRs in macula densa cells. Outer stripe OTR mRNA peaked with 4 days of estrogen treatment, and decreased to undetectable levels with 31 days of treatment of ovariectomized females. Estradiol's induction of outer stripe OTR mRNA expression was blocked by the antiestrogen, tamoxifen, but was not affected by high levels of circulating oxytocin. A role for OTRs in regulating renal function independently of adrenal steroids was suggested by findings that adrenalectomized males showed high levels of OTR transcripts in outer stripe proximal tubule and cortical macula densa cells after 5 and 10 micrograms/100g of estradiol. Consistent with specialized roles for OTRs during female reproduction, OTR transcripts could not be detected in renal tissues of peri-parturient females, at times when OTR mRNA levels were very high in uterus. OTR gene expression in macula densa cells reappeared 4-8 days into lactation and attained control levels by day 20. Physiological experiments showed that estrogen + oxytocin decreased plasma [Na+] levels in ovariectomized rats at a time when proximal tubule OTR expression is maximal. These data are consistent with 1) cell specific regulation by estrogen of renal OTR gene expression and 2) the possibility that OTRs may be important mediators of steroid-induced alterations in renal fluid and/or solute reabsorption. PMID- 8713984 TI - Functional properties of a cloned ovine endometrial oxytocin receptor cDNA. PMID- 8713985 TI - Structure/function studies of the human oxytocin receptor by site-directed mutagenesis. PMID- 8713986 TI - The rat oxytocin receptor. cDNA cloning and immunocytochemical localization in brain, pituitary, mammary gland and uterus. PMID- 8713987 TI - Histoautoradiographic detection of oxytocin- and vasopressin-binding sites in the amygdala of the rat. PMID- 8713988 TI - Neurone-astrocyte interaction. Inhibitory influence of neurones on astrocytic oxytocin receptor expression. PMID- 8713989 TI - In the rat kidney the specific oxytocin binding sites have different selectivity for vasopressin in the cortex and the medulla. PMID- 8713991 TI - Oxytocin and the anterior pituitary gland. AB - Release of oxytocin into the vicinity of the long portal vessels connecting the hypothalamus with the anterior pituitary gland and the presence of short portal vessels connecting the posterior lobe to the anterior pituitary established the potential for the peptide to act in a neuroendocrine fashion controlling the release of one or several adenohypophyseal hormones. Indeed, oxytocin receptors are present in the gland and numerous trophic effects of the peptide have been described, some with apparent physiological relevance. Thus under defined physiologic conditions a participatory role for oxytocin in the physiologic regulation of at least two hormones, prolactin and adrenocorticotropin, has been evidenced and there is developing credence to the concept that oxytocin may indirectly control gonadotroph function as well. Most recently, novel technologies have identified subpopulations of oxytocin receptive cells within a given cell type in the anterior pituitary gland and the concept of a modulatory effect of the nonapeptide on primary regulatory events has arisen. PMID- 8713990 TI - Natriuretic role of oxytocin in sodium homeostasis in rats. PMID- 8713992 TI - Synthesis and metabolism of oxytocin in late gestation in human decidua. AB - Several studies in the past few years have supported the hypothesis that oxytocin (OT) is synthesized in a paracrine system within the pregnant human uterus and that this paracrine system may be an important regulator of the timing of human parturition. Using ribonuclease protection assays, we have demonstrated a three fold increase in the rate of synthesis of OT mRNA in human decidua around the time of parturition. We also have shown that a similar increase in OT mRNA and peptide synthesis can be stimulated in vitro by physiological concentrations of estradiol. This increase is inhibited by concomitant use of the estrogen receptor (ER) blocker tamoxifen or by transcription inhibitors. Progesterone had little, if any effect. We also detected mRNAs for ER and progesterone receptor (PR) in amnion, chorion and decidua with the same relative tissue concentrations as OT mRNA. The concentrations of ER but not PR increased significantly around the time of labour onset. To determine if local OT concentrations may be regulated by changes in OT metabolism, we determined kinetic parameters for OT metabolism in decidua, chorion and placenta. [3H]tyrosyl-OT was used as substrate. Metabolites were separated using HPLC and identified using amino acid analysis and mass spectrometry. Metabolism in decidua and chorion occurred predominantly via a cytosolic post-proline endopeptidase and the activity was comparable to placenta. In microsomal fractions, cystine aminopeptidase activity predominated and placenta had significantly more activity than decidua and chorion. There were no changes in any Km or apparent vmax values around the time of parturition. These findings support the existence of a paracrine system within human decidua that involves sex steroids regulating synthesis of OT and that undergoes significant changes around the time of parturition. Changes in local OT concentrations are controlled by rates of synthesis rather than rates of metabolism. PMID- 8713993 TI - Pathways to parturition. AB - Oxytocin is secreted during parturition to stimulate myometrial contractions and birth. Prior to the start of labour, oxytocin neurones undergo changes to prepare for optimal secretion during labour. Thus, during late pregnancy oxytocin secretion is limited by endogenous opioid inhibition. This does not appear to act at the oxytocin nerve terminals in the neural lobe since they in fact become desensitised to opioid inhibition, responding less to either the general opioid antagonist, naloxone, or to the specific kappa-opioid agonist U50,488, and kappa receptor binding decreases. However, removal of opioid inhibition on oxytocin neurones by naloxone activates oxytocin cell bodies and there is an increase in the number of cells expressing Fos protein in the supraoptic nucleus. This action is mediated via mu- and not kappa-opioid receptors since norBinaltorphimine (kappa-antagonist) is ineffective. Endogenous opioids are likely to act pre synaptically on inputs to oxytocin neurones, especially those from the brainstem since naloxone potentiates the firing rate response of oxytocin neurones to intravenous cholecystokinin administration which acts via noradrenergic neurones. The endogenous opioid, beta-endorphin may be responsible for inhibition of oxytocin neurones as both the peptide content and its precursor proopiomelanocortin mRNA content increase in the arcuate nucleus during pregnancy, whereas expression of the co-localized opioids, prodynorphin or proenkephalin A, in magnocellular neurones does not alter. PMID- 8713994 TI - Gonadal steroid regulation of oxytocin and oxytocin receptor gene expression. AB - By Northern blot analysis and in situ hybridization, we have determined that, at term, the rat uterine epithelium represents a major site of oxytocin (OT) gene expression. OT mRNA levels increase > 150-fold during pregnancy and, at term, exceed hypothalamic OT mRNA by a factor of 70. By cryoultramicroscopy, OT immunoreactivity was localized to transport vesicles in the apical compartment of uterine epithelial cells. Estrogens (E) act as a strong inducer of uterine OT gene expression in vivo, and this effect is potentiated 7-fold by concomitant progesterone (P) administration. We have also cloned the rat OT receptor (OTR) gene and developed a polymerase chain reaction (PCR)-based assay to measure OTR mRNA. Whereas OTR mRNA is strongly induced by E, P does not potentiate but slightly attenuates the E-induced rise. However, E-induced OT binding is completely reversed by concomitant P administration, suggesting an additional post-transcriptional effect of P. The mechanisms of E-induction of the uterine OT gene remain unclear, inasmuch as the OTR gene promoter does not contain a classical estrogen response element (ERE). Moreover, transfection analysis of a 3.1 kb OTR gene promoter fragment linked to a luciferase reporter gene indicates that promoter activity is induced 5-fold by calcium ionophore A23187 but not by E. PMID- 8713995 TI - Oxytocin and the timing of parturition. Influence of oxytocin receptor gene expression, oxytocin secretion, and oxytocin-induced prostaglandin F2 alpha and E2 release. AB - Oxytocin receptor (OTR) gene expression was studied in various tissues of the reproductive tract of pregnant cows and compared with ligand binding activity. Myometrium, intercaruncular endometrium, caruncular endometrium, cotyledons and fetal membranes, as well as the uterine cervix of pregnant cows expressed the bovine OTR gene. Receptor concentrations, measured by ligand binding to crude microsomal pellets, were comparable to OTR mRNA signal strength in all instances indicating that the receptor protein formation is probably regulated at the transcriptional level. During bovine pregnancy OTR gene expression was initiated at different times depending on the tissue. The expression of the gene for OT peptide was not found in any of the bovine uterine tissues but was found in the corpora lutea at term and during parturition and then at relatively low levels. Therefore endogenous OT is derived almost exclusively from the pituitary during bovine pregnancy. OT secretion occurred in a pulsatile manner during pregnancy; a significant increase in pulse amplitude was observed during the last days before delivery and a large surge was associated with active labor and delivery. We postulate that the temporal order of OTR gene expression in the uterine and intrauterine tissues is a factor in the synchronization of the events that eventually lead to the onset of parturition. Because OT receptor mediates different actions in different tissues OT has multiple functions in the mechanism of parturition. The peptide initiates and maintains myometrial contractions, it stimulates release of PGF2 alpha from the endometrium and fetal membranes and, as demonstrated in this study, OT induces PGE2 release from cervical tissues in an OTR dependent manner. We conclude that in pregnant cows, OT participates both in the events that prepare the reproductive tract for birth and initiate the birth process. PMID- 8713996 TI - A proposed role for oxytocin in regulation of endometrial prostaglandin F2 alpha secretion during luteolysis in swine. AB - Pulsatile secretion of endometrial prostaglandin (PG)F2 alpha is stimulated by oxytocin (OT) during late diestrus in domestic ruminants (i.e., cattle, sheep and goats) and results in corpus luteum (CL) regression leading to the onset of a new estrous cycle. Pulsatile PGF2 alpha release is also responsible for CL regression in swine, but the stimulus for its secretion from the uterine endometrium is not known. We propose that OT binds to specific OT receptors (OTR) on the endometrium to stimulate phosphoinositide (PI) hydrolysis, thereby activating the inositol trisphosphate (IP3)-diacylglycerol (DAG) second-messenger system to promote pulsatile PGF2 alpha secretion. Exogenous OT administered to cyclic gilts during late diestrus (days 10-16) decreased interestrous interval in three of four experiments. However, OT did not promote CL regression in hysterectomized gilts indicating that the effect of OT was uterine-dependent. Circulating concentrations of 13,14-dihydro-15-keto PGF2 alpha (the major stable metabolite of PGF2 alpha) were increased (p < 0.01) 10 min after i.v. injection of OT on days 14 and 16 in cyclic gilts and on days 10-16 in pregnant gilts, but the magnitude of the response to OT on all days in pregnant gilts was markedly reduced compared to the response in cyclic gilts on days 14 and 16. Mean density and Kd of OTR detected on endometrium of cyclic pigs 15 days post-estrus were 29.2 +/- 5.5 fmol/mg protein and 1.59 +/- 0.23 nM, respectively. Density of OTR was correlated with OT-stimulated PI hydrolysis (r = 0.83, p < 0.05) and PGF2 alpha secretion (r = 0.87, p < 0.10). Endometrial IP3 was increased within 30 seconds after OT treatment and preceded the increase in PGF2 alpha release stimulated by OT. Endometrial PI hydrolysis and PGF2 alpha secretion were similarly increased by AIF4-(phospholipase C activator), but not by cholera toxin (adenylyl cyclase activator). Although OT binding to OTR could be displaced by lysine-vasopressin and lysine-vasopressin stimulated PI hydrolysis, lysine vasopressin did not stimulate PGF2 alpha release. Distinct receptors for OT and lysine-vasopressin on pig endometrium were confirmed by treatment with 100 nM OT + 100 nM lysine-vasopressin which stimulated PI hydrolysis more than 100-200 nM OT or lysine-vasopressin alone. These results support the hypothesis that OT stimulates phospholipase C to hydrolyze PI, yielding IP3 and DAG second messengers which promote endometrial PGF2 alpha release during CL regression in pigs. PMID- 8713997 TI - Parturition: activation of stimulatory pathways or loss of uterine quiescence? AB - Parturition results from the establishment of phasic regular uterine contractions. Contractility in myometrial smooth muscle is stimulated by an increase in intracellular calcium ([Ca2+i]) which activates myosin light chain phosphorylation leading to increased myosin ATPase activity and enhanced rate of acto-myosin cross bridge formation. G proteins play a pivotal role in smooth muscle activation and relaxation by coupling cell membrane receptors to effector enzymes and ion channels. G alpha(s) and G alpha(i) stimulate and inhibit adenylyl cyclase, respectively and control cAMP formation. G alpha(q) stimulates phospholipase C resulting in the formation of two second messengers: inositol 1,4,5-trisphosphate (InsP3) which releases Ca2+ from the sarcoplasmic reticulum, and 1,2-diacylglycerol which activates protein kinase C. The oxytocin receptor stimulates myometrial contractility by increasing [Ca2+i] through both pertussis toxin resistant (G alpha(q)) and pertussis toxin sensitive (?G alpha (i)) pathways. beta-Adrenoceptors and prostaglandin EP2 receptors promote relaxation via G alpha(s)-adenylyl cyclase. The concentration of myometrial oxytocin receptors is five-times higher in pregnant compared to non-pregnant myometrium but decreases in samples obtained during labour. When myometrial slices are challenged with oxytocin there is a rapid increase in InsP3 levels with a time course which is similar to the rise in [Ca2+i] provoked by oxytocin in cultured myometrial cells. The formation of InsP3 in response to oxytocin in myometrial tissue at term is similar in samples obtained before and after the onset of labour. G alpha(q) and G alpha(i) are expressed at similar levels in non-pregnant and in pregnant myometrium obtained before or during labour. By contract, G alpha(s) levels are higher in pregnant compared to non-pregnant myometrium and decrease in samples obtained during labor. These changes in G alpha(s) are paralleled by prostaglandin E2-induced adenylyl cyclase activity in the same tissues. Parturition may be the consequence of downregulation of pathways that favour uterine quiescence by increasing cAMP formation, resulting in a relative dominance of stimulatory receptors that increase InsP3/Ca2+ availability. PMID- 8713998 TI - Oxytocin signalling in human myometrium. AB - A physiological role for oxytocin in stimulating uterine contractions during labour is well accepted, but has not yet been well defined. Oxytocin activates phospholipase C to produce inositol 1,4,5-trisphosphate, which releases Ca2+ from intracellular stores. There is considerable evidence that G-proteins are involved in this signalling pathway. The objectives of the present study were to determine the mechanisms of action of oxytocin in human myometrium. We have measured the effect of oxytocin on the formation of inositol phosphates (InsPs) in cultured human myometrial cells labelled with [3H] inositol and on changes in intracellular free Ca2+ concentration ([Ca2+i]) in single cells using a dynamic calcium imaging system. Pertussis toxin was used to obtain information on the G proteins involved. Oxytocin induced InsPs formation and [Ca2+i] mobilisation in a concentration-dependent manner in human myometrial cells. Our data suggest that two distinct types of G-proteins are involved in the oxytocin response: one most probably a member of the Gq family (pertussis toxin-resistant) and another of the Gi family (pertussis toxin-sensitive). Using Western blotting, we have found that the pertussis toxin-resistant G-proteins alpha(q), alpha(11) and alpha(2), and pertussis toxin-sensitive alpha(i1), alpha(i2), and alpha(i3) are expressed in these cells. We have also detected the phospholipase C isoforms beta(1), beta(2) and beta(3) which are regulated by G-proteins, and phospholipase C isoforms gamma(1) and gamma(2), regulated by receptor tyrosine kinase pathways. However, oxytocin does not stimulate tyrosine phosphorylation in myometrial cells. Extracellular Ca2+ does not play a direct role in the activation of phospholipase C by oxytocin. Protein kinase C causes a strong inhibitory feedback on the oxytocin pathway: protein kinase C activators abolish the response to oxytocin while inhibitors potentiate it. Oxytocin responsiveness is upregulated by incubating the cells in the presence of oestradiol. This effect is reversed by the anti-oestrogen tamoxifen. Oestrogens exert their effects on the oxytocin pathway at a postreceptor level, possibly by affecting the expression of G proteins and/or phospholipase C isoforms. PMID- 8713999 TI - Mechanisms regulating oxytocin receptor coupling to phospholipase C in rat and human myometrium. AB - Oxytocin increases myometrial intracellular free calcium by promotion of calcium entry and release of calcium from intracellular stores. Calcium release from intracellular stores is secondary to an increase in phosphoinositide (PI) turnover and generation of IP3. We have explored the biochemical basis for the coupling of oxytocin (OT) to phospholipase C (PLC). Rat myometrial membranes contain PLC beta, gamma, and delta isoforms as well as the GTP-binding proteins G alpha(q) and G alpha(11). Oxytocin stimulates both GTPase and PLC activity in rat and human myometrial membranes. These data and available structural information suggest that the oxytocin receptor couples to PLC through a GTP-binding protein. In support of this hypothesis, an antibody generated against the specific C terminal region of G alpha(q) and G alpha(11) inhibits both the oxytocin stimulated GTPase and PLC activities. This inhibition is reversed by neutralization of the antibody with the antigenic peptide. The data indicate that the oxytocin receptor couples to PLC, presumably of the beta subclass, via interaction with proteins of the G alpha(q/11) subclass. In the nonpregnant, estrogen-primed rat, the stimulation of PI turnover by oxytocin is inhibited by the hormone relaxin and by pertussis toxin. The effects of both of these agents are mediated by the action of cAMP-dependent protein kinase. In plasma membranes, GTP-stimulated PLC activity can also be inhibited by treatment with protein kinase A. These data suggest that cAMP-dependent phosphorylation at a step involving GTP-binding protein/PLC coupling can exert a negative effect on the stimulation of IP3 formation by oxytocin and thereby affect contraction/relaxation in the myometrium. PMID- 8714000 TI - Desensitization of oxytocin receptors in cultured human myometrial cells. PMID- 8714001 TI - Activation of inositol phosphates in rat myometrium by bradykinin. PMID- 8714002 TI - Oxytocin receptors in human ovaries during the menstrual cycle. PMID- 8714003 TI - Quantitative polymerase chain reaction for determination of different oxytocin receptor mRNA expression in non-pregnant and pregnant uterine tissue. PMID- 8714004 TI - Detection of oxytocin receptor in endocervical scrapings during pregnancy. PMID- 8714005 TI - Gene expression of oxytocin receptor in human ectopic endometrial tissues. PMID- 8714006 TI - Luteotropic and luteolytic effects of oxytocin in the porcine corpus luteum. AB - The presence and the release of oxytocin (OT) by corpora lutea (CL) of a number of species (Wathes et al. 1986, Watkins and Choy 1988) including ruminants (Ivell and Richter 1984, Hirst et al. 1986, Rodgers et al. 1983, Sawyer et al. 1986), primates (Dawood and Khan-Dawood 1986, Khan-Dawood 1987, Maas et al. 1992, Khan Dawood et al. 1993), and the pig (Pitzel et al. 1984, Einspanier et al. 1991, Jarry et al. 1992) have been amply verified. Conflicting results concerning the effects of OT on steroidogenesis have been published; the peptide has been shown to be luteotrophic (Sawyer et al. 1986, Maas et al. 1992, Jarry et al. 1990), to have no effects (Rodgers et al. 1985) or to be luteolytic (Auletta et al. 1984, Auletta et al. 1988, Pitzel et al. 1988) and it appears that this confusion is only in part due to species differences but also the age of the luteal tissue seems to be of crucial importance for the understanding of the effects of OT (Schams et al. 1983, Wuttke et al. 1993, 1994). In the present contribution we will focus largely on our results obtained in the pig and where applicable, compare them with those obtained in other species. We will thus demonstrate that OT is released by luteal cells (Jarry et al. 1990, Einspanier et al. 1991, Jarry et al. 1992) and that luteal cells have OT receptors (Sernia et al. 1989, Pitzel et al. 1993a) which mediate the effects of the peptide on steroidogenesis. Finally, we will address the question whether OT is inhibitory or stimulatory to progesterone (P) and estradiol (E2) release, and we will come to the conclusion that OT is both luteotropic and luteolytic (Wuttke et al. 1993, 1994). The CL of all species investigated so far consists of two steroidogenic cell types. The so called large luteal cells stem from follicular granulosa cells and they appear to be barely responsive to luteinizing hormone (LH)/human chorionic gonadotrophin (hCG) but they are highly receptive to prostaglandin F2 alpha (PGF2 alpha) (Hansel and Dowd 1986, Pitzel et al. 1990). Furthermore, they appear to produce OT (Rodgers et al. 1983, Theodosis et al. 1986). The small luteal cells are believed to derive from the follicular theca cells (Hansel and Dowd 1986, Pitzel et al. 1990). They are LH-receptive but synthesize few, if any, regulatory peptides. In the last few years it has become increasingly evident that cells deriving from the white blood cell line are involved in processes such as ovulation and luteolysis. Of crucial importance for the understanding of luteolysis is the morphological observation that macrophages invade the CL at the time of luteal regression (Adashi 1990, Paavola 1977, Kirsch et al. 1981). PMID- 8714007 TI - Potential role of oxytocin in cell to cell communication in the corpus luteum. AB - Although oxytocin (OT) was identified in the human and primate corpus luteum (CL) over a decade ago, a definitive role for this peptide has not been elucidated. Several in vitro models have been utilized to examine the most obvious role for OT in luteal function, that of its effect on progesterone (P) production. Using dispersed cells in short term incubations, cultured cells and microdialysis procedures utilizing intact tissue, variable effects of OT on P production have been obtained. We therefore hypothesized that OT may have other role(s) in this tissue. The follicle cells remaining after a successful ovulation has occurred undergo rapid luteinization and the tissue is extensively remodeled to form the CL. At this time, in most species, two types of luteal cells have been identified based on their morphology, biochemistry and size. Both cell types produce P, however only one cell type responds to the long distance modulator LH. Since both cell types appear to be needed for the overall synthesis of P in adequate quantities required for the preparation of the endometrium "information trafficking" must occur between the two cell types. Our recent studies have shown the presence of gap junctions and connexin-43, E-cadherin, an adhesion molecule and ZO-1 protein associated with tight junctions in the baboon and human CL. The effect of OT on connexin-43 in the baboon CL has been examined. PMID- 8714008 TI - Oxytocin: a follicular luteinisation factor in the marmoset monkey. AB - The distributions of oxytocin (OT) and the oxytocin receptor (OTR) have been characterised in preovulatory follicles of the marmoset monkey and are described in relation to the process of luteinisation using a combination of in vitro and in vivo techniques, including immunohistochemistry and granulosa cell culture. Ovaries were collected in the periovulatory phase before and 22h after exogenous gonadotropin (hCG) treatment, but prior to ovulation. Before hCG treatment, OT immunoreactivity (OT-ir) was found mostly in granulosa cells of antral follicles, especially in the layers nearest the antrum. In contrast, oxytocin receptor immunoreactivity (OTR-ir) was observed principally in basal granulosa cells of antral follicles. Progesterone receptor (PR) and 3 beta hydroxysteroid dehydrogenase (3 beta HSD), markers for luteinisation, were negative for all ovarian tissues collected before hCG treatment. After hCG treatment, almost all granulosa cell layers in antral follicles stained positively for both OT-ir and OTR-ir, most prominently in preovulatory follicles and especially in the cumulus oophorus. This increase in staining was associated with an induction of PR and 3 beta HSD activity in the granulosa cells of preovulatory follicles. OT production by granulosa cells in culture was stimulated by the addition of hCG, but only in cultures derived from preovulatory follicles; whereas FSH was without any effect. Addition of extrinsic OT to the cultures elicited an increase in progesterone production only for the granulosa cells derived from preovulatory follicles. Together, the results of the in vivo and in vitro studies point to an involvement of OT in the process of leuteinisation in the marmoset monkey. PMID- 8714009 TI - A novel biological aspect of ovarian oxytocin: gene expression of oxytocin and oxytocin receptor in cumulus/luteal cells and the effect of oxytocin on embryogenesis in fertilized oocytes. AB - Recently, several authors have reported the existence of oxytocin (OT) in mammalian granulosa-luteal cells after ovulation. The purpose of this study was to examine the evidence for gene expression and localization of OT and OT receptor (OTR) in the cumulus cells surrounding the oocytes. Cumulus cells with mature oocytes were obtained from experimental and clinical in vitro fertilization-embryo transfer (IVF-ET) programs. OT and OTR gene expression was analyzed with reverse transcriptase-polymerase chain reaction (RT-PCR) and RT PCR/single strand conformation polymorphism (SSCP). OT gene expression was detected in mouse and human cumulus cells. The results of RT-PCR/SSCP showed that the structure of OT mRNA in cumulus cells was similar to that in the hypothalamus. Furthermore, OTR gene expression was clearly demonstrated in human cumulus cells, and a weak positive signal was observed in human oocytes. Immunocytochemical staining of OTR was clearly detected in human cumulus cells. The rate of mouse blastocyst development was significantly higher in the group cultured with OT than that without OT. These results are the first observations of simultaneous OT and OTR gene expression in cumulus cells, suggesting that ovarian OT might have some physiological role in the early stage of embryo development. PMID- 8714010 TI - Oxytocin and prostatic function. AB - It is now well established that oxytocin is present in the mammalian testis and there is growing evidence that the peptide plays a role in the male reproductive tract by both assisting sperm transport and modulating steroidogenesis. In the testis, oxytocin has been shown not only to modulate testosterone production but also to increase the activity of the enzyme 5 alpha-reductase which converts testosterone to dihydrotestosterone (DHT). The prostate is an androgen-dependent organ with DHT being the active steroid. Oxytocin is present in the mammalian prostate. We have shown in the rat that levels of the peptide can be regulated by androgens, prostatic oxytocin concentrations being decreased by testosterone and increased following castration or treatment with an antiandrogen. Oxytocin treatment increases 5 alpha-reductase activity in the prostate of healthy young rats but, unlike the testis, this rise in enzyme activity is only transient. We thus propose that a local feedback mechanism may act to control prostatic levels of DHT and hence prostatic growth. Benign prostatic hyperplasia (BPH) is a common disease which affects both men and dogs. The aetiology of the disease is complex but both DHT and aging are important factors. Oxytocin levels are raised in prostatic tissue from dogs with BPH and the increase in peptide is accompanied by increased 5 alpha-reductase activity. Preliminary findings also suggest that prostatic oxytocin levels are raised in tissue from men with BPH. These data lead us to suggest that oxytocin may be involved in the pathophysiology of the prostate gland. PMID- 8714011 TI - In vivo dynamics of oxytocin secretion by the ovine corpus luteum. PMID- 8714012 TI - Oxytocin in bovine and porcine granulosa cells. Regulation of secretion and biological effects. PMID- 8714013 TI - Luteotropic role of bovine luteal oxytocin in vitro. PMID- 8714014 TI - Pharmacological evidence for an oxytocin-like hormone in early pregnancy corpus luteum in sheep. PMID- 8714015 TI - Melatonin and serotonin directly regulate oxytocin, insulin-like growth factor-I and progesterone secretion by cultured human granulosa cells. PMID- 8714016 TI - Oxytocin regulates the secretory activity of human granulosa cells in vitro. PMID- 8714017 TI - Myometrial oxytocin messenger RNA is not increased at the onset of human labour. PMID- 8714018 TI - Effect of oxytocin on breast carcinoma cell growth. PMID- 8714019 TI - Expression of the oxytocin receptor in clinical human breast cancer tissues. PMID- 8714022 TI - Novel approach to oxytocin induction-augmentation of labor. Application of oxytocin physiology during pregnancy. AB - New information and understanding about the physiology of oxytocin (OT) have provided the bases of novel and rational approaches to the induction and augmentation of labor with OT. Based on a half-life of 8-10 min and time to steady state of 20 min, the interval between intravenous OT dose increments during uterine stimulation should be 20-30 min. Feto-maternal blood plasma gradients of OT concentration in spontaneous labor at term indicate a feto uterine flow of 2-4mU OT/min. Labor can be successfully induced in most women with such physiological doses (2-6 mU/min) of OT given intravenously. Direct and indirect evidence show that OT is released in pulses with the frequency and/or amplitude of the pulses increasing with or during spontaneous labor. Further OT interaction with its receptor renders it occupied and temporarily unavailable. We have therefore employed these observations to induce and augment labor with pulsatile intravenous administration of OT. In a prospective randomized study of 106 patients undergoing labor induction with pulsatile versus continuous administration of OT, the total dose, the average dose/min, and the peak or highest dose required/min were significantly reduced with pulsatile OT compared with continuous OT. These significant differences remained when controlled for parity, Bishop's score of cervical ripeness, and the number of days for induction. Similarly in a prospective randomized study on augmentation of labor by intravenous OT, we found the dose of OT for successful augmentation was significantly reduced with pulsatile OT compared with continuous OT. Thus, pulsatile OT not only is safe and requires less OT for uterine stimulation, but also constitutes a more physiological basis for uterine stimulation. PMID- 8714021 TI - Advances in the design of selective antagonists, potential tocolytics, and radioiodinated ligands for oxytocin receptors. AB - Despite intensive efforts over three decades in many laboratories, attempts to design peptide antagonists of oxytocin (OT) which are more selective for OT uterine receptors than for vasopressin (AVP), vasopressor V1a receptors, have met with only limited success. We will review the current status of the field and report on studies in our laboratories which have led to the design of highly potent non-selective and selective OT antagonists. Virtually all are more potent (2-6 fold) and a number are more selective (10-12 fold) than Atosiban, currently in clinical trial as a tocolytic agent. Many of these new published and unpublished OT antagonists are thus promising candidates for development as potential tocolytic agents for the prevention of pre-term labor. We also report on promising new radioiodinatable ligands for OT receptors. All the new OT antagonists are valuable new tools for studies on the physiological roles of OT and as probes for OT and AVP receptors. PMID- 8714023 TI - Potential use of oxytocin and vasopressin V1a antagonists in the treatment of preterm labour and primary dysmenorrhoea. AB - In order to study the involvement of oxytocin (OT) and vasopressin (AVP) in mechanisms of uterine activation and to clarify the therapeutic potential of antagonists to these hormones in preterm labour and primary dysmenorrhoea, studies of human uterine contractility in vivo and in vitro as well as measurements of OT and AVP V1a receptors were performed. Good correlations between OT receptor concentrations and effects on contractility were observed in both the pregnant and non-pregnant states, which indicates that OT acts specifically on its own receptor in the uterus. For AVP there was lack of such correlation which may suggest that this hormone influences both the OT and AVP V1a receptor sites. At the onset of labour both preterm and at term no marked increase in the OT receptor concentration was observed, but OT may still be involved in the initiation of labour, being produced locally in the uterus and not detectable in plasma. We observed a reduced OT receptor concentration in advanced labour and after OT infusion, which suggests that OT influences its own receptor. The AVP V1a receptor concentration and the effect of AVP on the uterus were about equal to those for OT, and the concentration of AVP V1a receptors also tended to decrease in advanced labour, observations which support an involvement also of AVP in the mechanisms of labour. In non-pregnant women AVP receptors as well as uterine effects of AVP in vitro and in vivo were about five times higher than those for OT, and the effect of AVP was increased premenstrually. This firmly supports an aetiological role of this peptide in the uterine hyperactivity of primary dysmenorrhoea. We have also shown that the analogue 1-deamino-2-D-Tyr (OEt)-4-Thr-8-Orn-OT, which blocks both the OT and AVP V1a receptor sites, given by intravenous infusion inhibits both preterm labour and dysmenorrhoea, and this is in agreement with our receptor and contractility findings. PMID- 8714024 TI - Progress in the development of oxytocin antagonists for use in preterm labor. AB - From a targeted screening effort and medicinal chemistry program, L-368,899 was selected as the first orally-active oxytocin (OT) antagonist to enter clinical trials. In animal studies, L-368,899 was shown to be a potent and selective OT antagonist and was orally bioavailable in rats, dogs and chimpanzees. L-368,899 was further shown to be a potent OT antagonist in pregnant rhesus and to inhibit spontaneous nocturnal uterine contractions. In Phase I human studies, L-368,899 was generally well-tolerated given intravenously and showed significant plasma levels after oral administration. In addition, L-368,899 blocked OT-stimulated uterine activity in postpartum women with a potency similar to that in the pregnant rhesus monkey. More recently, another structural series has been pursued, represented by L-371,257 [1-(1-(4-(N-acetyl-4-piperidinyloxy)-2 methoxybenzoyl)pip eridin-4-yl)- 1,2-dihydro-4(H)-3,1-benzoxazin-2-one]. L 371,257 exhibits high affinity (Ki, 4.6 nM) for human uterine OT receptors with high selectivity vs. human vasopressin receptors. In rat tissues in vitro, L 371,257 is a potent and competitive OT antagonist (pA2, 8.4) and, in vivo, blocks OT-stimulated uterine activity given both i.v. and intraduodenally. L-371,257 highlights the promise of this novel structural class. PMID- 8714025 TI - Enzyme immunometric assay (SPIE-IA) for oxytocin. PMID- 8714026 TI - Man and the chimaera. Selective versus neutral oxytocin evolution. AB - The oxytocin/vasopressin superfamily encompasses vertebrate and invertebrate peptides and therefore the ancestral gene encoding the precursor protein antedates the divergence between the two groups, about 700 million years ago. The preserved nonapeptide pattern indicates that both the precursor structures and the processing enzymatic machinery were greatly conserved to ensure the building of a specific conformation. Substitutions, which may be neutral or selective, occurred in precise positions. Virtually all vertebrate species possess an oxytocin-like and a vasopressin-like peptide so that two evolutionary lineages can be traced. Because a single peptide, vasotocin ([Ile3]-vasopressin or [Arg8] oxytocin) has been found in the most primitive Cyclostomata, a primordial gene duplication and subsequent mutations are assumed to have given rise to the two lineages. They started with vasotocin and isotocin ([Ser4,Ile8]-oxytocin) in bony fishes and culminated with vasopressin and oxytocin in placental mammals. Mesotocin ([Ile3]-oxytocin), found in lungfishes, amphibians, reptiles, birds and marsupials, appears as an evolutionary intermediate. The change from isotocin ([Ser4,Ile8]-oxytocin) into mesotocin ([Ile8]-oxytocin), can be observed in African and Australian lungfishes, species making the transition from bony fishes to land vertebrates. On the other hand the replacement of mesotocin by oxytocin can be detected in marsupials, particularly in the North-American opossum and the Australian Northern bandicoot that have both mesotocin and oxytocin whereas placental mammals possess only oxytocin. The invariability of this peptide in placentals can be explained by receptor-fitting selective pressure. In contrast to bony vertebrates in which neurohypophysial hormones revealed a remarkable structural stability, cartilaginous fishes displayed an unique oxytocin-like hormone evolution with variability and duality. Aside from vasotocin, in the subclass Selachii, rays have glumitocin ([Gln8-oxytocin]) and sharks possess two peptides: aspargtocin ([Asn4-oxytocin]) and valitocin ([Val8-oxytocin]) for the spiny dogfish, asvatocin ([Asn4,Val8]-oxytocin) and phasvatocin ([Phe3,Asn4,Val8] oxytocin) for the spotted dogfish. In the other subclass Holocephali, the chimaera (ratfish) has oxytocin, the typical hormone of placental mammals. Cartilaginous fishes used urea rather than salts for their osmoregulation and oxytocin-like hormones could have been relieved from osmoregulatory functions and able to accept many neutral variations. PMID- 8714027 TI - Structure and organization of the isotocin and vasotocin genes from teleosts. AB - Oxytocin and vasopressin are structurally related mammalian neurohypophysial hormones with distinct physiological activities. Homologues of these two hormones are found in all vertebrate groups except the primitive cyclostomes in which only a vasopressin-like hormone has been reported so far. Genes encoding the mammalian oxytocin and vasopressin hormone precursors exhibit a similar genomic organisation, and are closely linked (3.5 kb to 12 kb apart) in a tail-to-tail manner suggesting a common evolutionary origin. However, no linkage among genes of the vasopressin/oxytocin family has been demonstrated in any of the lower vertebrates. The Japanese pufferfish, Fugu rubripes, has been shown to have a very compact genome of about 400 Mb with very few repetitive sequences. We used this model vertebrate genome to analyse the genomic organisation of vasotocin and isotocin genes, the teleost homologues of vasopressin and oxytocin. We have cloned and mapped the vasotocin/isotocin locus in Fugu and obtained complete nucleotide sequences of the vasotocin and isotocin genes. The coding sequences of the Fugu vasotocin and isotocin genes are interrupted by two introns at identical positions, similar to their homologues in higher vertebrates. The deduced amino acid sequences as well as the coding sequences of the two genes show high homology in the neurophysin region. Furthermore, the two genes are linked in a tandem manner with an intergenic region of about 20 kb. Random sequencing has shown that there are at least two known vertebrate genes in the intergenic region. Genomic organisation of the Fugu vasotocin/isotocin locus suggests that the teleost vasotocin and isotocin genes have evolved from a common ancestor through tandem duplication and that this locus seems to have undergone a localised reorganisation during vertebrate evolution. PMID- 8714028 TI - Comparative aspects of oxytocin-like hormones in marsupials. AB - Unlike eutherian mammals which secrete only oxytocin (OT), marsupials secrete the typically reptilian mesotocin (MT) and/or OT as their oxytocic peptides. Our laboratories have been conducting research on various aspects of the roles of OT like peptides in three marsupials, the brushtail possum, the northern brown bandicoot and the tammar wallaby. By providing information on the functions of OT like peptides in these species we hope to provide some clues as to the evolution of neurohypophysial hormones in marsupials. Brain and peripheral distributions of OT-like peptides have been studied in the possum and bandicoot. As in eutherian mammals, OT-like peptides are distributed throughout the brain and are present in the testis, corpus luteum, prostate and adrenal glands. Studied on the regulation of release of MT into plasma in the possum show that it is regulated by similar mechanisms to OT release in eutherian mammals. OT receptors have been characterized and localized in the possum and the tissue distributions and pharmacological characteristics of the receptor are similar to both the sheep and rat OT receptors. The marsupial OT receptor shows no pharmacological specificity for MT over OT which is reflected in the similar potency of these peptides in eliciting contractions of the uterus of the tammar wallaby in vitro. MT seems to play an important but not essential role in parturition in the tammar. MT concentrations are increased immediately after delivery in the tammar but infusion of an OT antagonist before expected birth delays but does not prevent parturition. The presence of OT receptors in the marsupial mammary gland and the sensitivity of the gland to exogenous OT and stimulation of mesotocinergic neurones demonstrates that these peptides are important for marsupial lactation. Our data suggest that the presence of MT with or without OT in marsupials is a result of a neutral mutation rather than functional evolution. PMID- 8714029 TI - The hypothalamo-neurohypophyseal system in birds. AB - The hypothalamus is the central integrative structure for the neuroendocrine regulation of body fluid homeostasis. Despite many different modulatory endocrine and peptidergic inputs there are three main neuroendocrine 'channels' involved in osmoregulation: 1. corticotropin releasing hormone (CRH)-adrenocorticotropic hormone (ACTH)-adrenal steroids (aldosterone, corticosterone); 2. renin angiotensin; 3. arginine-vasotocin (AVT). In this paper the important role of the AVT synthesizing hypothalamo-neurohypophysial system (HNS) in the control of osmoregulation is described. Although the onset of AVT gene expression and peptide synthesis is as early as around day 6 of embryonal life in the chick, osmoregulatory function may not occur before the end of the second week of incubation. Rapid maturational processes during the last week of embryonal development lead to a 'close-to-mature' response of the AVT secretory system to physiological (osmotic) stimulation in the one-day-old chicken. Stable AVT plasma concentrations imply continuous secretion and/or degradation of this peptide in the blood of the chicken. Osmotic challenge causes not only an increase in plasma osmolality, plasma sodium and AVT concentration, but also AVT gene expression is upregulated. The AVT system participates in adaptational] processes during the perihatching period and its secretory activity depends on modulatory effects caused by suboptimal humidity during incubation. PMID- 8714030 TI - [Changes in the left ventricular diastolic function in acute myocardial infarction]. AB - Changes of left ventricular systolic function in acute myocardial infarction are well known. However, diastolic function may also have a role for patients' symptoms and prognosis. The purpose of this study was to evaluate changes of diastolic function non-invazively and to delineate the relation between systolic and diastolic function. 52 patients with acute myocardial infarction were examined by Doppler echocardiography within the first 48 hours and one week later. Systolic function was defined by left ventricular outflow velocity measurements, diastolic function was characterized by mitral inflow parameters and patterns. Four types of mitral inflow were determined: normal pattern, patterns suggesting prolonged relaxation, pattern suggesting elevated filling pressure and "normalized" pattern. In the last situation the influence of prolonged relaxation and elevated filling pressure equalized each others effect producing a "normalized" inflow pattern. Diastolic function improved in 18 cases and impaired in 9 pts, systolic function improved only in 11 cases and decreased in 28 pts by the time of the second examination. Worsening of diastolic function was always accompanied by worsening of systolic function. Despite improving diastolic function, worsening in systolic function could be observed in 8 cases. Monitoring of patients with acute myocardial infarction by Doppler echocardiography offers a possibility to select high risk patients with worsening left ventricular function for further closer follow-up. PMID- 8714031 TI - [Polipoid melanoma]. AB - Polypoid melanoma is a less known aggressive variant of nodular melanoma. It is characterised by earlier manifestation, exophytic growing, more frequent secundary alteration and whorse prognosis. On the basis of clinical and histological data of 14 patients, authors present characteristics of polypoid melanoma its differential-diagnosis and draw attention to the stressed importance of clinical diagnosis and treatment. PMID- 8714032 TI - [Incidence of post-transfusion hepatitis in Hungary 1987-1993]. AB - The authors describe the epidemiological characteristics of hepatitis cases occurred after administration of blood or blood-preparations in Hungary, based on data collected between January 1987 and December 1993. The epidemiologists of the public health network reported 868 acute posttransfusion hepatitis within this seven years period. The number of the cases decreased year by year, and in accordance with the rapid development of virological diagnostics the rate of cases with uncovered aetiology increased gradually. Nevertheless the aetiology of more than half of the reported cases (466 patients, i.e. 53.6%) remained unknown. The results of the examinations were negative in 167 cases (19.2%), and no etiological examinations were carried out in 299 cases (34.4%). Hepatitis A was reported in 17 cases (2%), hepatitis B in 129 cases (14.9%), whilst non-A, non-B hepatitis was diagnosed in 188 cases based on examinations with an experimental NANB antigen and antibody tests or by exclusion of hepatitis A and B infectious (21.7%); from 1991 67 cases (7.7%) were diagnosed by standard tests as hepatitis C, and Epstein-Barr virus infection was reported in 1 case (0.1%). During the seven years 11 patients of the 868 (1.3%) died in the acute phase of the illness. PMID- 8714033 TI - [Absence of unilateral pulmonary circulation, detected by perfusion scintigraphy of the lung, and its clinical significance]. AB - Single acute shortage of pulmonary circulation may cause a life threatening condition, its definitive lack--in consequence of developmental trouble- decreased loading and inclination to infections. It's recognition is most easy by non invasive isotope perfusion pulmonary scintigraphy. Frequency, diagnostic difficulties are a little bit known in Hungary, therefore retrospective investigations were performed by the authors. Examining 961 perfusion pulmonary scintigraphic material of a four year (1990-1994) period-among them that of 52 children-in 8 cases total perfusion-lack of one lung-side was found. Surveyng history of 5 children and 3 adults, besides conventional physical and X-ray suspicion-sings, attention is drawn to signs of non-invasive perfusion and aerosol inhalation pulmonary scintigraphy which are more suitable to demonstrate deviations. In the background of severe circulatory trouble there have been found in one infant grave bronchial stenosis, in 4 children--with different age- arteria pulmonalis agenesia at right, and in 1 young adult (with Fallot tetralogy), ateria pulmonalis agenesia at left. One of the other adults had bronchial carcinoma, and the other massive pulmonary embolisation. In their diagnostic schedule besides isotope investigations, importance of cardiologic echo-examinations, and the bronchoscopy has been stressed respectively. Catheterisation of the heart is important only in selected cases. PMID- 8714034 TI - [Diagnostic problems of Wilson disease]. AB - A family (three siblings) of Wilson's-disease is described. The authors review the pathogenesis, diagnostics, pathology and treatment of Wilson's-disease. The diagnostic difficulties are emphasised. The variety of liver lesions are demonstrated in the different grades of the disease. The importance of the early diagnosis is stressed. PMID- 8714035 TI - [Remembering Gabor Petri]. PMID- 8714036 TI - [The number of physicians in Hungary 1841-1989]. PMID- 8714037 TI - [Pre-symptomatic diagnosis of Huntington disease by polymerase chain reaction]. AB - The authors are reporting on the presymptomatic diagnosis of the autosomal dominant Huntington's disease applying the polymerase chain reaction for the first time in Hungary. The detection of the products yielded by polymerase chain reaction was carried out by silver stained polyacrylamide gel. This silver staining gave appropriate result without using radioactive technique. Cause of the disease is the greater number of CAG repeat sequence in the region of the Huntington's disease gene. The number of CAG repeats in Huntington patients is more than 37, while in not affected persons it is less than 34. Four members of two families each were examined and presymptomatic diagnoses were performed in two cases of both families. Normal alleles were detected in all cases. The tests supported the diagnoses of these patients suffering of clinically proved Huntington's disease. PMID- 8714038 TI - [Low doses of acetylsalicylic acid effectively inhibits thrombocyte aggregation after ischemic stroke]. AB - Platelet aggregation was examined in 43 patients after ischemic stroke and in 16 healthy subjects using multiparametric aggregation index (MAI). The value of MAI was significantly higher in stroke patients (3.15 in patients and 0.92 l/mumol in controls, p < 0.0001). Patients who had increased MAI (n = 26) were treated with a daily dose of 100 mg acetilsalicylic acid (ASA). Platelet activity was measured before and on the 7th and 28th day of treatment measuring three parameters: MAI, spontaneous dysaggregation and collagen induced aggregation. All 3 methods showed a significant decrease in platelet aggregation on the 7th day of treatment, but further changes were not found on the 28th day. Serum levels of thromboxane-A2 (TXA2) and prostacycline (PGI2) metabolites (TXB2 and 6-keto-prostaglandin-F1 alpha) were determined before and on the 28th day of treatment. The effect of 100 mg ASA per day proved to be selective: comparing the serum levels before and after treatment, a significant decrease of TXB2 concentration was found without changes in the concentration of 6-keto-prostaglandin-F1 alpha. Evaluating MAI and the value of dysaggregation might reflect ineffectiveness of antiplatelet therapy in patients not responding to a daily dose of 100 mg of ASA. For these patients the increase of the daily dose of ASA, or changing to another antiplatelet drug might be recommended. PMID- 8714039 TI - [Palliative treatment of large bile duct stones by endoscopic implantation of endoprosthesis in high risk patients]. AB - Endoscopic insertion of biliary stents is an established method in the palliative treatment of malignant bile duct stenoses. However, there is less experience concerning the long term effect of endoprostheses in patients with large bile duct stones. PATIENTS AND METHOD: Straight polyethylen endoprostheses of 12 Fr diameter were transpapillary placed because of large bile duct stones in 25 patients during a 5-year-period. The stents were inserted for transitoric decompression and for permanent treatment in 2 and 23 cases, respectively. Unsuccessful endoscopic stone removal (inc. mechanical lithotripsy) in high risk patients or extremely limited tolerability of patients during the endoscopic procedure were regarded as indications of stenting. The patients were followed clinically, biochemically and endoscopically for a period of 6 months to 5 years (mean 22.7 months). RESULTS: No early complication was observed. Late cholangitis occurred in three patients due to endoprosthesis dislodging and clogging in two cases and one, resp. All complications were successfully managed by endoscopic route. In further three patients elective stent exchange was performed. 8 patients died during the follow-up period, the cause of death was independent of gallstone disease in all cases. Without any stent exchange, 13 patients are symptom-free for 6-60 months. CONCLUSIONS: Endoscopic endoprothesis placement is a simple, cost-effective, relatively safe and well tolerable method for transitoric biliary decompression and for long term treatment in high risk patients with endoscopically not removable large bile duct stones. In contrast with malignant stenoses, in cases of bile duct stones elective exchange of the stent is not necessary, only clinical and biochemical follow-up suggested. PMID- 8714040 TI - [The level of use of statistical data in articles in the Orvosi Hetilap]. AB - The objective was to assess the level of use of statistics in the articles of the Orvosi Hetilap (OH). Eight clinical and 25 randomly selected general medical papers (published in 1992) were evaluated with the help of checklists used in the British Medical Journal (BMJ). The result was compared with the survey conducted in the BMJ (Gardner and Bond, JAMA, 1990, 263, 1355-1357). Two clinical studies were found suitable for publication without further revision. All the general papers needed revision and in 8 studies no changes appeared to be sufficient for publication. For the clinical and general articles, respectively, loglinear modelling showed a 4 and 6 times higher likelihood for satisfactory qualification in the BMJ than in the OH. In the OH, the quality of the analysis and presentation was rated the least satisfactory while the design was assessed the best among all checklist items. The main source of methodological flaws were either the lack of any statistical method or the inappropriate selection of them. The validity of the results published in the papers evaluated was often diminished by multiple testing or neglecting potential confounders. Contribution of statisticians appears desirable in planning and evaluating the investigation. Following the practice in the major medical journals, introduction of statistical review in the OH was suggested. In the view of the authors, reforms in medical education and research seem to be unavoidable for responding to the challenge of today's medical methodology. PMID- 8714041 TI - [Angiocentric T-cell lymphoma of the lung]. AB - Angiocentric T-cell lymphoma of the lung. The case history of a patient with primary angiocentric T-cell lymphoma of the lung having an unusually long survival period (> 10 years) is presented. Attention is paid to the possibilities of differential diagnosis that should be taken into account in the analysis of certain lymphocytic infiltrates of the lung. In accordance with relevant data of the literature, this case shows that pleiomorphic small cell T-lymphomas may have a protracted course, and the disease free periods repeatedly achieved in this patient by irradiation and chemotherapy are thought to be noticeable. Authors refer to some recent findings which may give new insights in the pathobiology of extranodal T-cell lymphomas, and result in recognition of new disease entities. PMID- 8714042 TI - [Problems in the pathogenesis, clinical picture and timely diagnosis of a diphtheritic lesion of the ENT organs]. PMID- 8714043 TI - [The physiological functions of the vestibular apparatus]. AB - Physiological functions of the vestibular apparatus were considered in terms of functional systems, permanent physiological activity and robot theories. A working hypothesis is assumed on vestibular receptor as a sensory detector of some functional systems of the body which enables creation of the internal coordinate system together with proprioreceptors of ocular, cervical, trunk and limb muscles, articular and tendinous receptors for adequate spacial perception via ocular and tactile analyzers; extrapolation of somatic responses. Vestibular reactions in disease and experiment are believed to be systemic responses primarily of functional and oculomotor systems. Isolation of vestibular afferentation from the total afferent flow is impossible already at the level of vestibular nuclei. Perspectives of further usage of rotatory and caloric tests for investigation of vestibular function are outlined. PMID- 8714044 TI - [The use of computer expert systems in studying the olfactory analyzer in the diagnosis of organic brain pathology]. PMID- 8714045 TI - [Positioning nystagmus]. AB - 256 patients with postural vertigo were examined clinically and neurootologically. Positioning nystagmus (PN) was detected in 209 (81.6%) of them. Postural vertigo arose as a result of inner ear diseases or traumas (n = 52), affection of cervical spine (n = 24), cardiovascular diseases (n = 97), organic impairment of the posterior cranial fossa (n = 20), endocrine disorders (n = 5), intoxication from chronic infection foci (n = 24), neuroses with vegetative dysfunction (n-34). PN emerged, as a rule, in patients having 2-3 of the above conditions. A genesis scheme of PN is outlined with regard to interaction of the main pathogenic factors influencing the vestibular system. Horizontal-rotatory or rotatory, vertical, diagonal, bilateral horizontal or multiple PN are characteristic for peripheral vestibular dysfunctions and central vestibular dysfunction, respectively. It is reasonable to perform position tests after assessment of the patients' nervous and cardiovascular systems, the condition of cervical spine. PMID- 8714046 TI - [Low-energy laser radiation in the combined treatment of sensorineural hearing loss and Meniere's disease]. AB - 59 patients with neurosensory hypoacusis and 45 with Meniere's disease underwent helium-neon laser intra- or supravascular radiation of blood. The treatment proved effective in acute neurosensory hypoacusis and Meniere's disease. In chronic neurosensory hypoacusis the effect was insignificant. PMID- 8714047 TI - [A model for the study of the ototoxic effects of drugs in an experiment]. AB - The authors compared sensitivity of the acoustic analyzer elements to ototoxic drugs in cat experiments. The model proposed assesses primary responses of the brain cortical acoustic zone (BCAZ) to sound stimulation. The drugs effect on the analyzer's periphery was determined from changes in bioelectric responses of the cochlea to sound stimulation (microphone potential, potential of the acoustic nerve action). The arguments are presented for feasibility of using primary BCAZ responses evoked by electric stimulation of thalamocortical radiation fibers coming from the medial geniculate body for examination of the drug effects on the analyzer's cortex. Parallel registration of the above bioelectrical reactions in the same animal for a long time makes it possible to compare sensitivity of different elements of the acoustic analyzer to ototoxic medication. PMID- 8714048 TI - [Upper respiratory tract function in workers in modern dye (cotton) manufacture]. AB - Clinical and functional examination of the upper respiratory tracts (URT) was performed in 240 employees exposed to novel active vinylsulfone dyes at cotton dye works. Those in contact with active dyes developed affections of URT mucosa leading to chronic subatrophic and catarrhal rhinopharyngitis occurring in dye exposed subjects 3 times more frequently than in the nonexposed. The severity of URT lesions depended on the occupation and length of service. Special tests provided evidence for early harmful action of the dye aerosols on URT in the majority of the examinees. PMID- 8714049 TI - [The characteristics of the membrane-destabilizing mechanisms in chronic suppurative and suppurative-polypous sinusitis in children]. AB - It is found that chronic purulent and pyopolypous sinusitis in children causes activation of lipid peroxidation and endogenic phospholipases in red cell membranes. There are shifts in structural and functional pattern of cytomembranes in inflammation focus (exudate from the sinuses). Membrane destabilization was maximal in pyopolypous sinus inflammation. PMID- 8714050 TI - [A method for studying the transport function of the ciliated epithelium of the nasal cavity mucosa in warm-blooded animals]. PMID- 8714051 TI - [Adoptive immunotherapy methods in ENT oncology]. AB - The search for methods providing selective action on some subpopulations of immune system cells is an important task of oncologic immunology. Adaptive immunotherapy [AI] may create new options in this field. AI was used as adjuvant in combined therapy of 180 patients with ENT cancer. The response was confirmed by correction of some immunological, cytological, biochemical, immunochemical, radioimmunological, biophysical and rheological parameters. Practical guides are presented on AI use at different stages of combined treatment. PMID- 8714052 TI - [Primary and delayed combined plastic repair of pharngoesophagostomies in patients with laryngopharyngeal cancer]. AB - Primary and delayed combined plastic surgery using massive musculocutaneous pedicle flaps composed of musculus pectoralis major, musculus sternocleidomastoideus, musculus trapezius in combination with cellulocutaneous and skin flaps provides appropriate plastic repair of total and subtotal defects of the pharynx and cervical esophagus after combined interventions for oropharyngeal cancer stage III-IV. Composite flaps warrant good and satisfactory functional and cosmetic outcomes in the majority of patients without additional hospital stay. PMID- 8714053 TI - [The use of pilastin for treating patients with chronic suppurative otitis media]. AB - Insertion of turundas covered with pilastin into the acoustic meatus alleviates exacerbation of chronic otitis media purulenta. As a result, disability does not last more than 2-12 days and was followed by a persistent remission and activation of immunity. Pilastin enhances proliferation and division of lymphocytes, increases blood levels of IgG and IgA. PMID- 8714054 TI - [The use of a combined prosthesis for the auditory ossicle chain of the author's own design in tympanoplasty]. PMID- 8714055 TI - [The use of autologous fibrin adhesive in septoplasty]. AB - Autologous fibrinous adhesive (AFA) was used in 40 patients with septal defects. It makes septoplasty more effective and reliable, is well tolerated, inexpensive because can be produced from autologous blood. PMID- 8714056 TI - [Characteristics of the course of temporal lobe abscesses in patients with multiple otogenous intracranial complications]. PMID- 8714057 TI - [Management of patients with chronic productive forms of sinusitis combined with a deviated nasal septum]. PMID- 8714058 TI - [Late seqelae of surgical treatment for ozena]. PMID- 8714059 TI - [A combination of exacerbated chronic suppurative epitympanitis and tick-borne encephalitis]. PMID- 8714060 TI - [A case of tuberculous meningitis erroneously interpreted as otogenous]. PMID- 8714061 TI - [Malignant paraganglioma (chemodectoma) of the larynx]. PMID- 8714062 TI - [The rating evaluation of student knowledge in a department of otorhinolaryngology]. PMID- 8714063 TI - [Gestational trophoblastic tumors]. PMID- 8714064 TI - [Laparoscopic surgery in gynecology. Experience at the Luis Castelazo Ayala Hospital]. AB - The purpose of the present study was to demonstrate the various applications of the laparoscopic surgery in the field of gynecology. 40 patients who were submitted to laparoscopic surgery were studied and divided into 5 groups accordingly to the surgical procedure: adhesiolysis, surgery of ovarian tumors, hysterectomy, surgery of the Fallopian tube and myomectomy. All the patients had a preoperative study and once completed they were selected for laparoscopic surgery. The laparoscopic procedure was able to solve the gynecologic pathology. The mean overall surgical time was of 66.8 min with a maximum time of 180 and a minimum of 30 min, accordingly to the groups the surgical mean times were as follows: 1) adhesiolysis 111 min 2) ovarian surgery 69 min 3) hysterectomy 113 min 4) tubal surgery 60 min and 5) myomectomy 58 min. There were no surgical complications and they all had a brief hospitalization period and a fast return to active life. All procedures were done with a bipolar cautery. We conclude that laparoscopic surgery is a useful resource for the resolution of the majority of the benign gynecologic pathology and when indicated in the proper way there is no rise in the morbid-mortality, and offers a short hospitalization period and a quick return to active life. PMID- 8714065 TI - [Post-cesarean necrotizing fasciitis. Bacterial synergism, or mixed infection of the soft tissues?]. AB - It has been demonstrated recently that most of the disemminated infections of the dermis, necrotizing fascitis included are due to a mix bacteria infection working sinergistically, demonstrated by clinical studies as well as in experimental animals. Treatment based on wide spectrum antibiotics and extensive debridement of the necrotic tissue performed by a team of several specialists have diminished mortality to 10%. We report a case of necrotizing fasciitis following a cesarean section in the absence of risk factors. We discuss risk factors, classification, etiology, diagnosis and therapy in light of the current knowledge. PMID- 8714066 TI - [HELLP syndrome. Critical state. Current concepts]. AB - The HELLP syndrome is an English acronym, for describing patients with Pregnancy Induced Hypertension (PIH), who also has hemolysis, elevated hepatic enzymes and low platelets, its presence is associated with a frequency of maternal mortality from 5 to 25% and when it is associated with hepatic rupture it increases to 35% and a perinatal one from 30 to 60%. It is present in 2 to 12% among women with PIH. Its etiology has not been elucidated completely, the base of its pathophysiology is an unbalance in prostanoid metabolism which conduces to generalize vasopasm with the subsequent endothelial damage and platelets activation. Sibai describes the diagnostic parameters of the syndrome. Its initial treatment is the hemodinamic stabilization of the maternal status and the evaluation of fetal well-being for decide whether immediate delivery is indicated. HELLP appears in the puerperium in 30% of the cases. There are both maternal and perinatal severe complications, and a recurrence risk of a 4 to 27%. We emphasize the importance of early detection of the syndrome which improves maternal-fetal prognosis. PMID- 8714067 TI - [Vaginal evisceration following radiotherapy and surgery for cervico-uterine cancer. Report of a case]. AB - A 60-year-old woman with past history of perineorrhaphy due to vaginal prolapse; squamous cell carcinoma of the uterine cervix stage IIB, treated with radiation theraphy at doses of 7500 rads administered as teletherapy and brachytherapy with Cesium 137. Ten weeks later a Piver II hysterectomy was performed, her posoperative morbidity was a vaginal abscess. Twelve years later, she developed a pulmonary metastasis. One year thereafter she had vaginal discharge of cetrinous fluid and prolapse of greater omentum through the vagina. She was treated by a midle exploratory celiotomy, primary closure of the vaginal defect and the pelvic floor was covered with an omental pedicle flap. The postoperative period was unremarkable. PMID- 8714068 TI - [Progressive systemic sclerosis and pregnancy]. AB - Nine cases with the association of systemic progressive sclerosis and pregnancy at Instituto Nacional de Perinatologia, from 1984 to 1994, are presented. All the patients were referred with the confirmed diagnosis by histopatological study. The variety CREST type was seen in three patients. Average age of patients was 29.2 +/- 5.4 years. Acute hypertensive disease of pregnancy; pre-term birth; and retarded intrauterine growth, were the most frequent complications. Most of the pregnancies were resolved by abdominal via, due to maternal and fetal indications. No maternal deaths. PMID- 8714069 TI - [Incidence and indication for cesarean section at the Central Military Hospital of Mexico]. AB - During the past two decades cesarean section rate has increased in recent years, various factors have been identified as responsible for increasing of this operation. The objective of this investigation was to determine the frequency, indications and complications of cesarean section performed at Hospital Central Militar (HCM). A prospective open study was performed during 2 years, 6977 pregnant women were included, of these, 5,105 patients (73.16%) had vaginal delivery and 1,872 (26.84%) cesarean sections were performed. The most frequent indications were failure to progress in labor (29.87%), repeated cesarean sections (22.86%) and breech presentation (9.61%). Endometritis was the major cause of positive morbidity (13.24%). Cesarean section rate had a significant increase of 11% in a 12-year period (1982 through 1994), almost of 1% annual. Attempt should make to halt the rise by marking recommendations as vaginal delivery after a previous cesarean section, breech deliveries. Also it is necessary to use appropriate intrapartum electronic fetal heart rate monitoring to avoid increased rates of unnecessary intervention. PMID- 8714070 TI - [Puncture of the dura mater, diagnosis and treatment in obstetrics]. AB - During the daily practice of Gynecoobstetrics there is a great relationship with anesthetic procedures. It is very important to know one of the most frequent complications of peridural blockade, as is accidental puncture of duramater. A review of this complication will be done, in order to help the physician to face this complication with an ample knowledgement, in order to be able to start opportune and adequate treatment. PMID- 8714071 TI - [Factors related to the recognition of alarm signals during pregnancy]. AB - INTRODUCTION: The medical literature has noted the relationship between perinatal morbidity and mortality and cultural factors for more than twenty years. During this time, however, rates of diseases such as hypertension, which complicate pregnancy and delivery, threatened preterm delivery and premature rupture of the membranes have decrease little. These conditions required other, complementary actions such as health education. The purpose of this research was to explore existing knowledge about warning signs during pregnancy in a population using third level medical facilities as the first step in designing an educational program. MATERIAL AND METHODS: We applied a 17 item multiple choice questionnaire regarding alarm sings during pregnancy to a group of 100 pregnant women. The sample was classified into six groups according to the age of the patient and the presence of aggregate risk. The analysis focused on the proportion of correct responses, applying different statistical tests according to the nature of the variables. RESULTS: In stepwise multiple regression, years of education explained 8.1% of the variance of correct responses, the number of pregnancies 5.5%, and gestational age 6.6%. The ANOVA of the complete regression was significant (F = 9.41; df = 99; p < 0.001) with a standard error of 2.23 ans a final adjusted r = 0.45. The number of consultations was positively associated with gestational age but had no relationships with correct responses. Groups with the greatest information deficits were adolescents and patients of adequate reproductive age with low reproductive risk. CONCLUSIONS: Patients with the least obvious alarm signs failed the most items in the alarm sing questionaire. The educational relationship between the physician and the patient should be more carefully explored. We are in agreement with other authors that educational level of patients is a determining to the attitude and knowledge of the population towards its own health. PMID- 8714072 TI - Rat epididymal sperm exhibit on dithiothreitol treatment in vitro quantifiable differences in patterns of light scatter, uptake of 14C-iodoacetamide and binding of ethidium bromide to DNA. AB - The extent to which chromatin of rat caput (CAP), corpus (COR), cauda (CAU) spermatozoa undergo condensation and compaction is known to be a function of progressive increase in the formation of inter- as well as intra-protamine disulphide bridges during their transit through the epididymis. Relative compaction undergone by the nuclear chromatin of these sperm populations was studied by monitoring their susceptibility to in vitro decondensation induced by varying concentrations (0, 0.01, 1, 5, 10, 50 mM) of disulphide reducing agent, dithiothreitol (DTT) after an initial exposure to 0.01% papain. Following this treatment and staining with the nucleic acid specific fluorochrome, ethidium bromide (EB), it was observed that irrespective of the epididymal region from which they were collected, spermatozoa exhibited DTT dose-dependent (a) increase in nuclear size as seen under fluorescence microscopic examination, (b) decrease in flow cytometrically quantifiable light scatter parameters--forward scatter (FSc, 'nuclear size') and side scatter (SSc, nuclear 'granularity'), (c) increase in individual cell EB binding when analyzed by DNA flow cytometry, and (d) increase in thiol specific 14C-iodoacetamide (14C-IA) uptake. The decrease in both FSc and SSc occurring in spite of actual increase in nuclear size has been attributed to increase in translucency of spermatozoan nuclei consequent to decondensation. The FSc, SSc and EB bindability were studied by monitoring both the channels of maximal cell concentration detected in the flow cytograms as well as by digitally quantitating the numbers of cells within specific channels (1-64, 65-128, 129-192 and 193-256) of the flow cytogram. The latter indicated a measure of the variability in the response of populations of sperm within each sample to DTT induced decondensation. At any given concentration of DTT, especially between 5-10 mM, the differences observed between sperms of different regions were consistent and significant (P < 0.01-P < 0.001), maximal changes being shown by CAP and minimal by CAU sperm, COR sperm appearing in between. The effective concentration of DTT required to elicit 50% of maximal (i.e. that exhibited by CAP sperm when taken as 100%) effect (ED50) varied significantly among CAP, COR and CAU sperms for each of the parameters studied (P < 0.01-P < 0.001). It is concluded that the differences observed among the three epididymal sperm populations are due to differences in the extent of susceptibility to decondensation in vitro and that this is dependent upon the variation in the -S-S content of their chromatin during different stages of epididymal transit. All the parameters used (with the exception of fluorescence microscopy) can be quantified and as all of them show a similar dose dependency to DTT treatment, any one of these parameters can be conveniently used to determine the mature/immature status of the sperms voided. Application of such a method to determine the quality of sperms voided by man appears feasible. PMID- 8714073 TI - Assessment of developmental retardation and abnormality of in vivo produced preimplantation embryos in rat. AB - In most mammals studied, a substantial numbers of preimplantation embryos are believed to be lost in vivo. In vitro, embryos develop slowly and lose viability. Hence, there is a need to assess the extent and cause of embryonic loss both in vivo and in vitro. In this study, we assessed the quality of in vivo produced ovulation products/embryos, recovered on days 1-5 pregnancy, from naturally bred wistar rats. From day 1 pregnant rats (n = 24), 226 ovulation products were recovered which included 52% (117) unfertilized oocytes and empty zonae with/without cell debris (UFO-EZ:CD) and 48% (109) 1-cells. Flushings of day 2 rats (n = 27) contained 229 ovulation products, consisting of 70% (160) 2-cells and 30% (69) UFO-EZ:CD. Flushings of day 3 rats (n = 27) had 23% (56) 2-cells, 6% (15) 3-cells, 23% (57) 4-cells, 1% (2) 5-7 cells, 2% (5) 8-cells and 45% (112) UFO-EZ:CD, total being 247. Flushings of day 4 rats (n = 28) had 193 ovulation products comprising of one morula, 45% (86) 8-cells, 5% (9) 5-7-cells and the rest were 4-cells (2), 3-cells (2), 2-cells (1) and 48% (92) UFO-EZ:CD. Day 5 flushings (n = 27) had 202 ovulation products which included 13% (27) morulae, 17% (34) early, 36% (73) mid and 2% (5) late blastocysts; additionally, 4-cells (1), 8-cells (2) and 30% (60) UFO-EZ:CD were also recovered. On day 4, embryos (8 cells) migrated from the oviduct to the uterus. When pregnant rats (n = 25) were allowed to term, only 15 females (60%) delivered pups (128) with variable litter size (2-12). These results indicate that 56% (619/1097) of recovered rat preimplantation embryos are of expected developmental age with a mixture of asynchronously cleaving embryos. The remaining 44% (478) is comprised of 38% (417) UFO-EZ:CD and 6% (61) abnormal and developmentally retarded embryos, which are unlikely to produce viable pups at term. PMID- 8714074 TI - Radioimmunoassay of angiotensin-II: methodology and standardization. AB - The study was planned to set up and standardize the radioimmunoassay of Ang-II and to validate the procedure in terms of precision, sensitivity, specificity and recovery. The application of the developed assay was studied in normal healthy volunteers and in patients of renovascular hypertension (RVHT) and renal hypertension (RH). Synthetic human Ang-II was coupled to BSA by carbodimide condensation to get the hapten carrier conjugate which was injected in rabbits to raise the antibodies. The titre of 1:250 showed significant binding (56.79%) and was used for the assay. The sensitivity of the assay was 2 pg/ml and cross reactivity with analogues of Ang-II was minimum. Mean Ang-II levels in normal subjects was 16 +/- 3.6 pg/ml. In patients of RVHT and RH, the peripheral blood Ang-II levels were found to be 876 +/- 8.6 and 108 +/- 2.3 pg/ml respectively. In patients of unilateral RVHT, renal vein Ang-II levels of the affected side were significantly higher than the unaffected side (P < 0.001). The results indicate that unextracted samples can be successfully utilized to estimate Ang-II levels. PMID- 8714075 TI - Isolation and characterization of intraepithelial lymphocytes from rat small intestine. AB - A modified procedure for isolation of intraepithelial lymphocytes (IEL) from rat small intestine was developed which makes use of a protease inhibitor-phenyl methyl sulfonyl fluoride (PMSF) in the isolation medium. Yield and viability of IEL isolated in presence of PMSF were significantly higher as compared to those isolated in absence of PMSF. IEL isolated in presence PMSF demonstrated a significantly higher proliferative response to concanavalin A (con A) compared to those isolated in absence of PMSF. The natural killer cell activity of IEL population isolated in presence of PMSF using YAC-1 lymphoma cells as targets was also significantly higher compared to those isolated in absence of PMSF. Phenotypic analysis using monoclonal antibodies specific for rat T lymphocyte subsets revealed that majority of IEL were cytotoxic/suppressor phenotype with a minor subset of helper/inducer phenotype. The method described yielded a population of IEL which is suitable for further functional studies in vitro. PMID- 8714076 TI - Comparative evaluation of anticonvulsant activity of calcium channel blockers in experimental animals. AB - Pentylenetetrazole (PTZ)-induced convulsions and the maximal electroshock (MES) seizure test were employed to study the anticonvulsant effects of nifedipine (2, 3.5 and 5 mg kg-1), flunarizine (10, 20 and 40 mg kg-1) and diltiazem (10, 15 and 30 mg kg-1). Nifedipine and flunarizine prolonged the latent period and reduced the mean duration of PTZ induced seizures. They also reduced the severity of convulsions and the number of deaths due to PTZ significantly. Nifedipine was more potent in this regard (P < 0.01). All these drugs prolonged the latent period and reduced the duration of tonic extensor phase of MES seizures in a significant manner. Flunarizine was most potent in this test. Complete protection from tonic extensor phase was observed in 10-50% animals pretreated with nifedipine and flunarizine in a dose dependent manner. The response of diltiazem was weak in both these tests. It is concluded that all three calcium channel blockers possess an important but different anticonvulsant effect and their significant clinical use can be made while keeping in view the characteristics of their pharmacological action. PMID- 8714077 TI - Synergistic effect of alcohol and nicotine on glycosaminoglycan metabolism in rats. AB - Oral administration of alcohol along with nicotine decreased all the glycosaminoglycan (GAG) fractions except hyaluronic acid in aorta and liver of rats. Decreased activity of the enzymes involved in the biosynthesis of precursors of GAG and increased activity of many of GAG hydrolysing enzymes indicate decreased biosynthesis and increased degradation of GAG. Sulphate metabolism in liver was also significantly altered by administration of both alcohol and nicotine showing considerable decrease in the concentration of sulphated GAG. PMID- 8714078 TI - Studies on glucoamylase produced from Aspergillus awamori (NRRL-3112) and their effect on saccharification of potato starch. AB - Efficiency of an enzymatic starch saccharification process depends not only on the activity of the glucoamylase but also its purity. About 96.82% unwanted proteins present in 2-day old culture broth of A. awamori NRRL-3112 (grown in MYGP medium) were separated by precipitation with ammonium sulphate which was followed by dialysis. More than 80% activity of the glucoamylase was recovered. Three protein fractions (A, B, C) were identified using gel permeation chromatography. Fractions A and B showed comparatively higher glucoamylase activity than fraction-C. Moreover, fraction-B showed no product inhibition. The optimum temperature and pH of the purified enzyme (fraction-B) were 60 degrees C and 4.0 respectively. The saccharification efficiency of potato pulp was more in case of using purified glucoamylase (fraction-B) as compared to that of crude enzyme. PMID- 8714079 TI - Tamoxifen increases the proliferation of human endometrial stromal cells in in vitro. A model for evaluation of endometrial hyperplasia. AB - Tamoxifen given for breast cancer therapy, has a complex and an unclear action on the endometrium. A large number of literatures has attributed the proliferous changes in the endometrium caused by tamoxifen (Tam). No report has appeared on the endometrial cellular changes induced by Tam. The present study shows a significant (P < 0.001) increase in the proliferative activity due to Tam in endometrial stromal cells over control and estradiol (E2). This in vitro model is useful for the study of the hyperplasic effect of Tam at the cellular level. PMID- 8714080 TI - Quantitative assessment of amphetamine induced behavioural changes in rhesus monkeys, Macaca mulatta. AB - Graded dose of amphetamine (AMP; 1-4 mg/kg, im) induced suppression of approach, contact, body jerk, grooming and food forage, hypervigilance (checking), stereotyped behaviour and oral hyperkinesia in rhesus monkeys. Pretreatment with haloperidol--a dopamine receptor blocking agent (0.04 mg/kg, im)--significantly suppressed the AMP induced hypervigilance, stereotypy and oral hyperkinesia but was unable to reverse other AMP induced behavioural effects in the monkeys. Haloperidol (0.01-0.04 mg/kg, im) per se produced decrease in social and solitary behaviour and marked cataleptic posture. The results suggest that dopamine may be playing an important role in the mediation of many AMP induced behavioural changes in primates. PMID- 8714081 TI - Laboratory and field evaluation of an insecticidal trap for houseflies. AB - Several substances/food baits and admixtures thereof were tested in a perspex trap, treated with insecticide K-othrine@0.01 mg/cm2. Bioefficacy for housefly trapping was evaluated. Combination of ingredients was found to be more effective than individual ingredients. A combination of some easily available and cheap ingredients were employed for field trials. The admixtures of milk powder, dog biscuit, jaggery and banana peeling was found very effective. The trap efficacy including persistence of the toxicant K-othrine was observed for more than 6 months. Observations on number of flies trapped and utility of such a trap in actual field situation are discussed. PMID- 8714082 TI - Association of a cucumber mosaic virus strain with mosaic disease of banana, Musa paradisiaca--an evidence using immuno/nucleic acid probe. AB - Virus causing severe chlorosis/mosaic disease of banana was identified as a strain of cucumber mosaic virus (CMV). Association of CMV with the disease was established by Western immunoblot using polyclonal antibodies to CMV-T and slot blot hybridization with nucleic acid probe of CMV-P genome. PMID- 8714083 TI - Effect of quaternary benzo[c]phenanthridine alkaloids sanguinarine, chelerythrine and fagaronine on some mammalian cells. AB - The effects of benzo[c]phenanthridine alkaloids sanguinarine (SA), chelerythrine (CHE), fagaronine (FA) and their dihydroderivates were tested on human leukocytes and lymphocytes, rat peritoneal mastocytes and primary cultured hepatocytes. The cytotoxicity of SA and CHE on hepatocytes is dose (35-100 microM) and time (1-3 h) dependent. Both alkaloids decrease chemiluminiscence of leukocytes and inhibit a creation of active E-rosets. The degranulation of mastocytes is inhibited only by CHE. Dihydroderivates of SA and CHE did not display any effect on studied cells, dihydrofagaronine exhibits a hepatotoxicity after 3 h. PMID- 8714084 TI - The effect of soil fertilization on the formation and the amount of cannabinoid substances in Cannabis sativa L. in the course of one vegetation period. AB - The study followed the effect of the soil fertilization on the growth of plants and on the formation and the amount of extractible substances as well that of two main cannbinoid substances (CBD, delta-9-THC) in the Czechoslovak variety of hemp, Rastislavice, cultivated in Czechoslovakia for fibre production in the course of the vegetation period of 1988. In fourteen various vegetation stages of the plant growth, the samples of the plant tops, cultivated on five fields with different soil fertilization, were collected and analyzed in the dried state. PMID- 8714085 TI - Chitin assay in aspergillus fumigatus. AB - Our study is an attempt to make the determination of chitin possible for large amount of samples with high sensitivity and reproducibility. These requirements are fulfilled if using acid hydrolysis to release glucosamine from chitin, evaporating the solution to remove hydrochloric acid, dissolving it in distilled water to obtain optimal concentration and assaying it fluorimetrically. The described method was used for determination of chitin in spores of Aspergillus fumigatus. PMID- 8714086 TI - Effect of the fluorescent brightener Rylux BSU on the cell wall chitin content in Basidiobolus ranarum. AB - In Basidiobolus ranarum an artificial cell dimorphism was found if cultivated in presence of Rylux BSU previously. We have found an increase of glucosamine content in purified cell walls of Basidiobolus ranarum grown in presence of Rylux BSU in SGA. The relative increase in glucosamine content did correspond with the increase of Rylux BSU present in SGA. The results are discussed with the conclusion that not only the chitin synthesis but also the mechanisms of polarized growth are influenced if Basidiobolus ranarum is cultivated in presence of Rylux BSU. PMID- 8714087 TI - Development and validation of an in-house radioimmunoassay for erythropoietin. AB - A sensitive radioimmunoassay (RIA) for the determination of erythropoietin (Epo) in blood serum and amniotic fluid has been developed and validated. The assay uses a polyclonal antibody obtained in rabbits immunized with recombinant human Epo by one of the authors. Tracer and second antibody are commercially available. With 100 microliters of sample, the assay can detect 3 U/l. The reproducibility of the assay compares well with commercial Epo RIAs. The normal values estimated in a group of blood donors are slightly higher than those reported for various RIA determinations. The clinical usefulness of the assay was demonstrated by observing the expected increases or decreases of Epo levels in various clinical states known to alter Epo production. The assay meets all requirements of a routine clinical assay at significantly reduced costs, which makes it particularly suitable for use in research applications. PMID- 8714088 TI - Expression of p53 protein in tissues and organs of human and mouse embryo. AB - The expression of p53 was demonstrated in the cells of some tissues in the various stages of human embryo development. The distribution of this protein correlates in some cases with sites of higher proliferative activity as demonstrated by PCNA/cyclin detection. It is necessary to prove whether the p53 protein participates in the regulation of cell division, or it is the aberrant expression of unfunctional p53 which is induced in sites of higher proliferation by feed back regulatory mechanism. PMID- 8714089 TI - New alternatives for the treatment of fistulas in Crohn's disease. AB - The authors refer on the combination of the administration of artificial nutrition with enterohormone Somatostatin in a group of 16 patients with Crohn's disease (CD), complicated by the occurrence of fistulas. The sole administration of total parenteral or enteral nutrition led in each case to a significant reduction of secretion from the fistulas, and in two cases to their complete closing. Apart from this, the nutritional status of the patients improved. The administration of Somatostatin i.v. in the form of the preparation Stilamin led to a further reduction of secretion from the fistulae. Complete closing of the fistulae due to a combination of total parenteral nutrition (TPN) or total enteral nutrition (TEN) and Stilamin, was achieved in only 2 patients, but the combination use of tissue sealant Histoacryle with the treatment resulted in a closure of the fistulas in a further 2 patients. Altogether the application of this treatment resulted in closing 6 fistulas (37.5%). The authors consider that the above mentioned methods can open new prospectives, but this requires further experience. PMID- 8714090 TI - Epidemiology of bronchial asthma in children in Olomouc district. AB - We investigated prevalence of bronchial asthma in children during the period 1973 1992 in Olomouc district. Bronchial asthma was detected in 839 children, which is 1.2% of all children's population. It has risen 4 times since 70ties. 80% of the disorders were detected during 1st-4th year of life. PMID- 8714091 TI - [Solving crisis situations in psychiatry]. AB - Provision within the area of medical care has been made for psychological and social intervention in some crisis situations. A Help line telephone service called "Linka nadeje" was established in 1965 and a crisis intervention centre in 1989. Client characteristics, and an analysis of telephone calls made during the course of three ten year periods (N = 27,268 total phone calls) are compared with 10,021 contacts and 1,781 first admissions during a five year period at the crisis centre. Data showed that both institutions help meet current crisis and in the long term they should both reduce the risk of psychological crisis, reduce the incidence of suicide, they should also help prevent both primary and secondary illness and supply sufficient information on the surrounding area. PMID- 8714092 TI - A comparison of various methods for cooling the kidney. AB - Various methods for achieving preoperative renal hypothermia were tested and evaluated. A kidney immersed in an oil bath solution at 4 degrees C resulted in the highest rate of contact hyperthermia at a determined safe temperature of cooling medium. The acquired basic temperature curve served as a model for the comparison of other methods tested. The results showed that perfusion hypothermia is most rapid but this method is limited by the number and state of the renal arteries and displays all the risks associated with cannulation of arteries. Covering the kidney with crushed ice led to a sufficiently rapid decrease in temperature but lack of regulation carries the danger of renal damage caused by excessive temperature reduction. The disadvantages of these methods were eliminated using a device developed and tested by the authors. PMID- 8714093 TI - Kidney temperature measurement. Thermistors for temperature measurement. AB - During a study of the protective effects of cold on kidney cells under conditions of interrupted blood circulation, it was found that attention must be paid to the temperature of kidney tissue in order to prevent cell damage caused by an excessively reduced temperature. The authors correspondingly developed an instrument for the measurement of kidney tissue temperature where thermal sensors located in a needle are able to measure deep temperature in live tissue under conditions of external hypothermia. Thermal sensors were also placed in a hypothermal coil for contact measurement of surface temperature of a cooled kidney and in a duct for supply of cooling medium into a coil. Construction of these devices is described and basic the characteristics of the thermistor and thermocouple sensors for temperature measurement are presented. After evaluation of advantages of the tested sensors, indications regarding their application are given. PMID- 8714094 TI - A revised instrument for the regulated external cooling of the kidney. AB - The authors describe the instrument they have developed for the regulated external cooling of the kidney. This consists of a cooling thermostat fitted with a circulating pump, coils of their own construction for the surface cooling of the kidney and equipped with sensors for the measurement of kidney surface temperature, tube connectors, and a sensor for the measurement of the cooling medium temperature. A cooling liquid is supplied by means of a circulating pump from a thermostat through elastic tubes into the flow channels of the cooling coils. The temperature of the coolant is recorded by means of a thermistor sensor built into the system of tubes while the temperature of the kidney surface itself is monitored by a sensor installed in a coil. This permits the regulation of both the rate and depth of temperature reduction in order to prevent possible damage to the kidney caused by excessive temperature reduction. The efficiency and safety of this instrument were verified both experimentally and clinically. PMID- 8714095 TI - A revised coil for the external cooling of the kidney. AB - The authors describe a coil they have developed for contact peroperative hypothermia of the kidney. This new type of coil eliminates the disadvantages of existing cooling coils, namely slow thermal transfer from kidney to cooling agent due to imperfect contact between the coil and the concave surface of the kidney, use of an inadequately conductive material and thermal losses from the outer surface of the coil. Special features of the coil described here include its construction to match the contours of the kidney, isolation of external shell to reduce thermal losses and continuous monitoring of renal tissue to reduce the danger of overcooling. PMID- 8714096 TI - Protection of kidney cell against ischaemic damage by decreased kidney temperature. AB - At normal body temperature, kidney cells are highly sensitive to hypoxia. However, a bloodless operative field is advantageous in some interventions on the kidney. Kidney cells can be protected against the damage that might occur once blood flow has been interrupted pharmacologically or physically by reducing their temperature. The authors compare several methods for achieving renal hypothermia, including a method based on a device they developed themselves to eliminate defects found in existing equipment. The thermal curve obtained when the kidney is immersed in an oil bath cooled to 5 degrees C served as a model for other methods. Using the revised coil developed by the authors, renal hypothermia was achieved safely and efficiently. A combination of pharmacological and physiological methods is preferrable however in order to potentiate the effect of decreased temperature. This method was tested experimentally and then successfully applied in clinical practice. PMID- 8714097 TI - The initial experience with doppler diagnosis of scrotum disease abstract. AB - Blood flow parameters in the scrota of 83 men with acute and chronic infra scrotal disease were compared with values from a sample of 70 healthy controls, using doppler ultrasonography. The results confirm the validity of this method in the diagnoses of scrotum disease when combined with other methods of examination. Technical difficulties are discussed. PMID- 8714098 TI - [Benefits of percutaneous interference sonographic performance in kidneys]. PMID- 8714099 TI - Contribution to the clarification of heterogeneous foci within the prostate. AB - The determination of the nature of heterogeneous foci within the prostate is up till now insufficient. With the help of a standard method using a scanner with a changeable plane of cut and frequency of 7.0 MHz, 292 patients were examined with an enlarged or standard size of prostate and with morphologically proved prostate cancer. A high level of sensitivity and quite a low specification of examinations were found out. To distinguish the kind of heterogeneous foci within the prostate is of decisive importance for the carcinoma diagnostics. In 152 patients, the following characteristics were evaluated: the size, shape and contour of the focus, its localisation, echogenicity and homogeneity of echostructure. 70 patients were examined using a method evaluating the vascularisation of a tumour with CFM and the frequency analysis according to Furier. The results of examinations performed with the help of the above mentioned methods yielded some new experience, however, they did not contribute to the qualitative determination of heterogeneous intracapsular foci within the prostate. Their character can be defined only by punctual or aspirational biopsy. PMID- 8714100 TI - New possibilities of miniinvasive treatment of esophageal stenoses. AB - We report our first very good experiences with the treatment of inoperable, especially malignant stenoses of the esophagus, with perorally inserted metallic self-expanding stents. During the period since August 1993 up to now, we have introduced 17 stents in 15 patients. We have compared the introduction of stents with other modalities of palliative treatment of esophageal stenoses, as actinotherapy, contact or focused laser, palliative by-pass surgery, classic surgical or endoscopical stenose-pertubation. In the last place for the patient stands the dismal nutrition-gastrostomy. From this point of view, the miniinvasive treatment with metallic stents is very successful, easy, well tolerated by the patients, and practically without complications. The therapeutical effect, elimination of severe swallowing-discomfort, allows especially to patient with malignancies to live terminal months of life with a relatively maximum quality of deglutition. PMID- 8714101 TI - Mediastinitis due to perforation. AB - The oesophageal perforation is a begining of tragedy. We present a group of patients with serious complication-mediastinitis after operations of the oesophagus or after instrumental diagnostics and instrumental treatment. PMID- 8714102 TI - Professor Emil Starkenstein and the clinical pharmacology. PMID- 8714103 TI - [Ophthalmology at a generational change]. PMID- 8714104 TI - [Endocrine exophthalmos]. AB - The basedowian exophthalmy is a self-immune originating affection, in which the antigen remains still unknown. It may appear besides the Basedow disease, evoluating in a total independent manner. The clinical diagnosis is done on the basis of small ocular characteristic signs: exopthalmy, oculomotor deficit, optical neuropathy, corneo-conjunctival alterations, being of course supplemented with the complementary endocrinological and immunological explorations. The endocrine exophthalmy represents a serious complication because of the visual risks it implies, and afterwards because of the implied esthetical prejudice. A spontaneous amelioration is possible, but the treatment often proves to be necessary. It will be medical, local and general, and also orbitar radiotherapy and surgical treatment. PMID- 8714105 TI - [The pathogenesis of diabetic dyschromatopsia]. AB - Essential data about the physiology of colour vision are reminded, with a stress layed on the possible contribution of the rodes in perception of blue and green colours. The main features of the diabetic dyschromatopsia are reviewed, together with their pathomorphological support and the two existing theories regarding its pathogenesis: vascular origin versus neuronal origin of the defect. The contribution of the photocoagulation to these alterations is also discussed. PMID- 8714106 TI - [A family with glaucomatous excavation but without glaucoma]. AB - The authors present the case of a family consisting of eight members (belonging to three generations). Among them there are three cases with increased glaucomatous excavation (C/D 0.7-0.8) but normal visual field and ocular blood pressure. Their clinical tracking, till the age of 16, excepts the possibility of glaucoma in these cases, only a genetically transmitted abnormality being implied. PMID- 8714107 TI - [The ocular hypotonic effect induced by an angiotensin-converting enzyme inhibitor (captopril)]. AB - The investigation aims at analysing in healthy and glaucoma subjects the effects of an angiotensin conversion enzyme inhibitor (Captopril) on intraocular pressure, local tolerance, influence on systemic circulation and to ascertain (or suggest) the mechanisms inducing this pressure lowering. The study is prospective, masked, in acute test (10 days) and includes a series of 14 healthy subjects and 18 patients with open angle primitive glaucoma. Following numerous experimental and clinical tests, Captopril isotonic solution 2.5% administered in ocular instillation 4 times/day proved to be the most appropriate. RESULTS: Captopril solution 2.5% lowers intraocular pressure both in the normal subjects (average relief of 6.5 mm Hg) and in the glaucoma patients (8.9 mm Hg); hypotonic effects lasts for 6-8 hours; local tolerance is good; has no influence on systemic arterial pressure; tonography with constant Pt shows a 12% increase of C (insufficient to explain such a relief in intraocular pressure). The hypotesis that pressure lowering occurs (partially) by the stimulation of prostaglandin activity and consequently by improved uveoscleral drainage is suggested. CONCLUSIONS: Captopril solution 2.5% administered topically lowers intraocular pressure without adversely influencing systemic and local circulation; local tolerance is good; the mechanisms of ocular hypotonizing effect are unknown. PMID- 8714108 TI - A gliacentric view of the human retina. AB - The structure of retina was historically described using light microscopy histological sections. This description placed prime importance on neural elements and their interconnections is establishing retinal structure. Of the 10 retinal layers, 7 are neural elements. This neurocentric description is, however, not the only way to view the retina. After considering various pathological conditions, we believe that an alternative description of the retina exists, based on glial cells. The gliacentric view may better enhance our understanding of vascular pathologies of the retina. In contrast to the classical view of a 10 sheet neurocentric structure, we propose a gliacentric structure composed of two sheet. This model is supported by the embriological development of the retina, originating from two neuroepithelial sheets (the two layers of the optic vesicle). These neuropithelial layers later develop into two monostratified stheets composed of epithelial cells with the following characteristics: a basement membrane at the basal side, villi at the apical side and intercellular junction complexes at the lateral aspects of the cells. The neural network of the retina lies embedded within these sheets. This gliacentric model has not only theoretical implications but bay also explain why various pathologies of the retina are limited to one compartment rather than extending the entire thickness of the retina. PMID- 8714109 TI - [Biometry in cataract surgery with an implant]. AB - In cataract surgery, the replacement of the opaque lens with an artificial lens is the ideal correction. For the present, the intraocular lens power determination is an essential part of the preoperative evaluation of an eye. The paper presents a short history about the development of implant power calculation, the main formulas used in present, the precise choice of postoperative refraction, the factors affecting accuracy of implant power calculation and some preoperative considerations (patients considerations, intraocular lens characteristics considerations). The final part contains some conclusions about the methods of implant power calculation. PMID- 8714110 TI - [Exudative uveitis, chronic labial herpes and interstitial pneumonia in an HIV positive child]. AB - The paper presents the case of a 6 and a half year-old child, with marked hipotrophy in height and weight (15 kg weight, which make a deficit of about 6 kg). The right eye presented pupilar sinekies, loaded vitreous and two white yellow exsudative coroidal placards. One of them was inter-maculo-papilar having 4DP in diameter, difuse margins and aspect, the other was infero-macular. The right eye vision was only 2/50 n.c. The child also presented cronical labial herpes and interstitial pneumony. The ELISA test (+) confirmed the HIV suspicion. The paper insisted on the herpetical ethiology of the corio-retineal exsudative lesions. PMID- 8714111 TI - [Chronic ischemic ophthalmopathy in Takayasu-Onishi disease]. AB - The cronical ischemical ophthalmopathy represents an ocular clinical syndrome with well-defined clinical particularities. It is the effect of the decrease of the perfussion pressure at ACR level, during a long time period, caused by the stenosis of the aortic arch and his main branches. The paper presents a clinical observation of cronical ischemical ophthalmopathy having in ethiology the unspecific aortitis or the Takayasu-Onishi diasease. It is extensively discussed the ocular clinical syndrome and the particularities of the Takayasu-Onishi disease. PMID- 8714112 TI - The relative distribution of retinal and choroidal blood in the human retina. AB - In this paper a model is presented to explain the relative distribution of retinal and choroidal blood to the human retina. According to this model the retina between the inner limiting membrane and the outer limiting membrane is metabolically controlled and nourished by the retinal vasculature, through Muller cells. The retina between the outer limiting membrane and Bruch's membrane is nourished by the choroidal vasculature, through the retinal pigment epithelium. Muller cells have the ability of metabolic uptake from the interphotoreceptor space as demonstrated in pathological situations suchs as central retinal artery occlusion, when both the Muller cell and the photoreceptor nuclei survive. This demonstrates an overlap existing between the two distributions. Neural elements in the retina depend on Muller cells and RPE for metabolic support. It is only reasonable to expect that the cleavage plane between the two blood supplies is the outer limiting membrane, the outer limit of the Muller cell. In this model both the photoreceptor nuclei (the outer nuclear layer) and the foveal retina are assumed to be supplied by retinal. and not choroidal, vasculature. One consequence is that after long standing retinal detachment the photoreceptor outer segments degenerate (since they become deprived of choroidal blood supply) but the outer nuclear layer, nourished by the Muller cell, continues to survive. PMID- 8714113 TI - [Accuracy in the measurements of the glaucomatous papilla]. AB - The authors review different methods for quantifying papilae's modifications in glaucoma. For each method the possible errors are discussed. The conclusion is that C/D evaluation based only on biomicroscopical measurements is acceptable in everyday practice, the nervous fiber ring's measurement being not yet accurate and simple enough to be widely applied. PMID- 8714114 TI - [The ocular manifestations in Felty's syndrome]. AB - The Felty syndromme is a rare clinical form of the rheumatoid poliarthritis, which involves, besides the articular manifestations specific to the latter, other manifestations such as: splenomegalia with hypersplenism, poliadenopathy, ulcerations at the low leg level and skin pigmentation. In some cases, the Felty syndromme may be associated with ocular modifications: episcleritis, nodular scleritis, corneal ulcer, Sicca syndromme, iridocyclitis, retineal vasculite. A clinical observation of Felty syndromme associated with ocular modifications is presented. Some aspects regarding the nosologic classification of Felty syndromme, its clinical and ethiopathogenical particularities are discussed; the accent is put on the immunopathological component. The ocular modifications particularities, their diagnosis and the therapeutical aspects are presented. PMID- 8714115 TI - [Epithelial-cystic nevus of the conjunctiva (Parinaud's dermo-epithelioma)]. AB - The epithelio-cystic nevus of the conjunctiva, initially improperly denominated by Parinaud dermo-epithelioma, is a dysplasia circumscribed at the level of bulbar conjunctiva, characterized by its forming of epithelial cysts which develop in a base layer made of nevoid cells. This affection evolves by forming epithelial cysts mostly muciparus ones, when the tumor is sited rather far from the limbus, where multiple chaliced cells are to be found in the epithelium, and as simple epithelial cysts, mostly in the epidermis, when it is sited near the limbus where chaliced cells are scarcer. In this condition the setting of the epithelial compound takes the form of full lobules. The nevoid compound represents the capital element which establishes the diagnosis. The epitheliocystic nevus of conjunctiva is a benign dysembryo with excessive development of epithelial cells in a basic layer of nevoid cells. This affection is benign, but its morphologic diversity and its many denominations give rise to periodical debates. PMID- 8714116 TI - [Marginal ectasia of the cornea resolved surgically]. AB - The paper presents the case of a patient with Terrien's marginal degeneration. The moment of the exam, the corneal degeneration was in an advanced stage, being associated with secondary ectasia. The gutter developed as a results of the stromal progressive thinning was already circumferential. The only efficient treatment in this situation is an annular lamellar keratoplasty. We present the mais operatory moments, and also the main intra- and postoperatory incidents and complications. In spite of the fact that this operative procedure was already mentioned in literature, there are no certain proves; so, we are able to consider the achievement of the annular lamellar keratoplasty unique for this moment. PMID- 8714117 TI - [The secondary implant: yes or no?]. AB - The paper presents the arguments for and against the secondary implant. It is considered that the best results were obtained with anterior chamber flexible implant, excepting the young pacients, for who the "iris-chip" lens is recommended. The minimal interval between the primary and secondary implant is unanimously estimated to be one year. The diminishing of risks for this kind of intervention rely on the correctness of the pacients selection. This implies a careful previous clinical and paraclinical analysis, as well as patients antecedents examination. PMID- 8714118 TI - [The ophthalmological echograph]. AB - The ophthalmological echographical device is presented, its principles and functionning are reviewed in order to allow a better understanding of the functional limits. This is useful also for the process of selection from a large variety of existing variants and offers. PMID- 8714119 TI - [Orthopoxvirus genome]. PMID- 8714120 TI - [Polymorphism of enzymes involved in drug metabolism: gene structure and enzymatic activity]. PMID- 8714121 TI - [Malaria parasite DNA and RNA changes connected with acquired resistance to chloroquine]. PMID- 8714122 TI - [Intragenomic polymorphism of ribosomal RNA genes from human chromosome 13]. PMID- 8714123 TI - [Determination of the nucleotide sequence of the part of the Mycoplasma gallisepticum genome, containing the rpoB gene, during the use of the Bal-pTM method for obtaining sequential deletions of the sequenced fragment]. PMID- 8714124 TI - [Expression of fragments of Venezuelan equine encephalomyelitis virus E2 protein in a system based on phage T7 RNA polymerase]. PMID- 8714125 TI - [Construction of a large-scale physical map of the human genome. I. Preparation of the NotI-"binding" clone library]. PMID- 8714126 TI - [Transcription factors C-EBP and AP-1 modulate the promotor activity of the regulatory region of human p53 gene]. PMID- 8714127 TI - [Analysis of the 3'-terminal region of the tick-borne filament-like virus substantiates its affiliation with the shallot X-virus group]. PMID- 8714128 TI - [Design of secretion vectors for Escherichia coli]. PMID- 8714130 TI - [Calculation of the probability of connection in a ring of a short DNA fragment with proper curvature and proper axial torsion]. PMID- 8714129 TI - [Modeling DNA hydration using the Monte Carlo method. Interaction of adenines with water in a nonglycoside A-tract grove stabilizes the B'- conformation]. PMID- 8714131 TI - [Turning speed and stability of a native structure in "random" and "edited" chains]. PMID- 8714132 TI - [Differences in recognition of peptides of the MHC H-2Kb molecule between cytotoxic T-lymphocytes, memory cells, and T-suppressor cells, specific for alloantigen H-2Kb]. PMID- 8714133 TI - [Interaction of metastazin I with the heavy chain of nonmuscle myosin]. PMID- 8714134 TI - [Protein engineering of uridine phosphorylase from Escherichia coli K-12. Study of the role of cysteine residues in enzyme function]. PMID- 8714135 TI - [Thermodynamic analysis of the interaction of ethidium bromide with the deoxytetraribonucleotide 5'-d(GpCpGpC) from proton magnetic resonance data]. PMID- 8714136 TI - [Effect of sequences flanking the "-10" homology region of the spc-promoter from the starting point, on the interaction with Escherichia coli RNA polymerase]. PMID- 8714137 TI - [Processing results of a univariate fluorescence analysis of human chromosomes by flow cytometry]. PMID- 8714138 TI - [Interaction of uracil-DNA-glycosylase from human placenta with single-stranded deoxy- and oligoribonucleotides and their complexes]. PMID- 8714139 TI - [Apurine/apyrimidine endonuclease from human placenta. Recognition of apurinized DNA by the enzyme]. PMID- 8714141 TI - Cluster of cases of clinical cholera due to Vibrio cholerae 010 in east Delhi. AB - A total of 514 samples of acute diarrhoeal stools received over a period of four months yielded 315 isolates morphologically and biochemically resembling V. cholerae. Out of 315 isolates, 223 (70.8%) were identified as V. cholerae 01, 20 (6.4%) as 0139 and 42 (13.3%) as 010. Thirty (9.5%) isolates did not agglutinate with any of the available antisera. All V. cholerae 010 isolates showed complete homogeneity in their biochemical and physiological properties. This strain appears to be closely related to El Tor biotype of V. cholerae 01, since it was positive for some of the tests used for identification of El Tor. The ability of strain 010 to grow in the presence of 6 per cent salt provides it the status of an important environmental pathogen. Acquisition of some virulence genes from El Tor vibrios by this strain 010 appears to be one of the mechanisms involved in the emergence of this serogroup. PMID- 8714140 TI - [Interaction of primers with mutant forms of human immunodeficiency virus reverse transcriptase]. PMID- 8714142 TI - Outbreak of food poisoning due to Salmonella paratyphi A var durazzo (2,12:a:-) in Yavatmal (Maharashtra) in May 1995. AB - Thirty three patients presented with acute diarrhoea and vomiting, 12-24 h after consuming vegetarian food. Twenty three patients developed high grade fever, with two patients developing complications. Salmonella paratyphi A var durazzo was isolated from 12 faecal samples. This serovar of Salmonella as a causative organism of food poisoning is unusual. PMID- 8714143 TI - Development of an antigen capture enzyme immuno assay using genus specific monoclonal antibodies for detection of Chlamydia in clinical specimens. AB - An antigen capture enzyme immuno assay (EIA) for Chlamydia antigen detection was developed using polyclonal rabbit immunoglobulins against C. trachomatis, a genus specific antichlamydial murine monoclonal antibody (IgG) against major outer membrane protein (MOMP) antigen of C. trachomatis and a commercial anti mouse IgG immunoglobulin conjugated with HRPO. The test was evaluated against a direct immunofluorescence assay (DFA). Conjunctival specimens from 178 patients with follicular conjunctivitis and cervical specimens from 82 patients with cervicitis were tested for Chlamydia antigen detection by both the tests. Chlamydia antigen was detected in 69/178 (38.76%) and 68/178 (38.20%) of the conjunctival specimen by using EIA and DFA tests respectively. It could also be detected in 24/82 (29.27%) and 22/82 (26.83%) of the cervical specimens by EIA and DFA tests respectively. The sensitivity of the EIA test was 92.64 per cent and 86.36 per cent for the conjunctival and cervical specimens respectively against the reference DFA test. The specificity of the EIA test was found to be 94.54 and 91.66 per cent respectively against the reference DFA test. PMID- 8714144 TI - Development of malathion resistance in Culex quinquefasciatus Say (Diptera: Culicidae). AB - A malathion resistant colony of C. quinquefasciatus was developed in the laboratory. LC50 and LC90 for larvae were calculated at every generation and the values were 0.3 ppm and 1.13 ppm for first generation and 61.09 ppm, 136.3 ppm for 25th generation respectively. The fold increase in LC50 and LC90 were 2036 and 2726 folds respectively. Cross resistance against propoxur and chlorpyrifos showed 6.64 and 6.52 fold and 600 and 720 fold increase in their LC50 and LC90 values respectively. Triphenyl phosphate (TPP) and piperonyl butoxide (PB) were used as synergists and TPP indicated proportional decrease in LC50 and LC90 values while not much change was observed with PB. No change in biotic potential (larval hatchability, adult emergence and male and female ratio) between susceptible and malathion resistant colonies was observed. PMID- 8714145 TI - Insecticide susceptibility of Phlebotomus papatasi to organochlorine, organophosphate & carbamate compounds in some arid areas of western Rajasthan. AB - Insecticide susceptibility status of P. papatasi to organochlorine, organophosphate and carbamate compounds has been estimated in Pali and Barmer districts of Rajasthan. Tests revealed that this species was resistant to DDT but susceptible to dieldrin, malathion, fenitrothion and propoxur. Efficacies to these compounds at 50 and 95 per cent levels have been estimated by probit analysis. The LC50 values for both DDT and dieldrin, were found much lower than those reported from other parts of the country. Heterogeneity of the response was found highly significant [X2 = 43.8(3)] in case of propoxur only. PMID- 8714146 TI - Reversible non enhancing lesions without focal neurological deficits in eclampsia. AB - Nine patients with eclampsia, were subjected to computerized tomographic scan (CT scan) of the head to ascertain the changes in the brain that accompany seizures and encephalopathy of eclampsia. Only those patients who had a seizure within the past 24 h were included in this study. None of the patients had any focal neurological deficit. Six of the nine patients had abnormal findings on CT scan consisting of multiple non enhancing hypodensities in the cerebral white matter. One patient also had subependymal haemorrhage over the lateral ventricle. These changes were found to have disappeared when the CT scan was repeated on the seventh day. There was no correlation between the mean arterial blood pressure or the number of seizures and the presence of hypodensities in the brain. These findings suggest that subclinical changes in the form of reversible hypodensities and rarely bleeding can occur in eclampsia even when patients have no focal neurological deficits. It appears that these lesions represent focal areas of cerebral oedema, secondary to failure of autoregulation of cerebral blood flow. PMID- 8714147 TI - Valvular dysfunction in uraemia. AB - Regurgitation of the pulmonary, mitral, tricuspid and aortic valves have been observed frequently in chronic renal failure (CRF) and dialysis patients. Two dimensional, M mode and doppler echocardiography were performed on 35 CRF patients and 37 end stage renal failure (ESRD) patients on maintenance haemodialysis. Though structurally normal, valvular dysfunction was noted in 50 per cent of the patients with renal failure. Mitral regurgitation was the commonest abnormality, occurring in 36.1 per cent of the patients. Calcification of the valve was observed in only 5.6 and 16.7 per cent of CRF and dialysis patients respectively. Multiple regression analysis underscored the large contribution of diabetic status in the development of valvular dysfunction. Though end systolic volume was higher in patients with valvular abnormalities, the ejection fraction was well preserved. However, follow up studies are required to assess the significance of the functional valvular regurgitation on the cardiac function of the patients. PMID- 8714148 TI - Immunohistochemical study of the expression of human groEL-stress protein in human nervous tissue. AB - Monoclonal antibody (ML-30) directed against 65 kDa stress protein of mycobacteria, is shown to identify human cellular protein homologous with the groEL heat shock protein in many prokaryotes. Immunohistochemical survey of nervous tissue, both central and peripheral, from patients dying of various inflammatory, degenerative and neoplastic conditions and from experimental animals, using this antibody showed punctate granular staining of the cells to a variable degree. The astrocytes showed strong immunolabelling. The normal neurons and oligodendroglia stained variably, while abnormal neurons were darkly labelled. Ependymal cells showed apical granular positivity. The ubiquitinated inclusion bodies in amyotrophic lateral sclerosis, Alzheimer's disease and Parkinson's disease were not recognised by the ML-30 antibody. In diseased and stressed nervous tissue from experimental animals, the expression of the ML-30 recognisable stress protein was variable. The epitope recognised by ML-30 was found stable in postmortem tissues collected up to 36 h after death and processed for paraffin sectioning, after fixation in formalin for many years. Enhanced expression of the human groEL stress protein homologue in mammalian nervous tissue following various forms of stress may play a role in modulating the extent of tissue damage by autoimmune mechanism because of its high immunogenic nature and constitutive presence in the cells. PMID- 8714149 TI - Genetic & cultural determinants of high-density lipoprotein cholesterol & serum triglycerides among Marwaris of Calcutta. AB - A genetic epidemiological study of serum lipid and lipoprotein levels was conducted among families of Marwaris residents in Calcutta. A total of 210 families, comprising over 100 individuals, were studied. Analyses were performed to estimate the genetic and environmental effects on the determination of high density lipoprotein cholesterol (HDL-C) and serum triglycerides (TG). Familial correlations for HDL-C and TG were estimated: parent-child and sib-sib correlations were found to be significant. Spouse correlations were not significant. Correlations between environments of siblings were significant. Genetic analysis of data on HDL-C and TG performed under a path model, taking genetic transmission and possible environmental associations among family members into account, indicated that lipid and lipoprotein levels adjusted and standardized for age, gender, education, occupation and disease status are primarily determined by genetic factors. The effects of environmental factors were also significant, although in comparison with genetic factors these effects were much smaller. The estimated genetic heritability for HDL-C was approximately 80 per cent, while that for TG was approximately 55 per cent. The genetic effects and environmental effects were not significantly different between adults and children. PMID- 8714150 TI - The anti-inflammatory activity of some sulphonamides in albino rats. AB - Clinically equivalent doses of dapsone, sulphasalasine and sulphamethizole in albino rats showed significant (P < 0.05) anti-inflammatory activity in carrageenan and cotton pellet induced inflammation. Their activity was comparable to that of aspirin (200 mg/kg) and was confirmed by granuloma histology. Further, these compounds also showed significant (P < 0.01) analgesic activity which was comparable to that of aspirin. The ulcer index for sulphamethizole was comparable to that of control animals, whereas dapsone and sulphasalazine showed significant ulcerogenicity (P < 0.01). Other sulphonamides like sulphadiazine, sulphanilamide, sulphamoxole and cotrimoxazole did not show significant anti inflammatory and analgesic activities. PMID- 8714151 TI - [Clostridium difficile: an increasing nosocomial pathogen]. PMID- 8714152 TI - [Frequency of the clinical manifestations of Lyme borreliosis in Spain]. AB - INTRODUCTION: There are differences in the clinical manifestations of Lyme borreliosis (LB) in different parts of the world. The aim of this work was to analyze its clinical manifestations in Spain. METHODS: We evaluated by ELISA and IFI the sera of 1,500 patients with clinical suspicion of LB between January 1987 to February 1993. Spanish criteria of LB (amplified CDC criteria of LB for epidemiological purpuse) were used. Clinical and serological data were evaluated with the patients' physicians and other etiologies were reasonably excluded in accepted cases of LB. RESULTS: Sixty-four patients of 138 with a positive serology (46%) met the LB criteria. Neurological manifestations were presented by 40 patients (62.5%) (in control group 23%, p < 0.05) cutaneous lesions by 20 patients (31%), articular manifestations by 18 patients (28%) (in control groups 56%; p < 0.05) and cardiac manifestations in two. Cutaneous manifestations included 17 erythema migrans, 2 acrodermatitis chronica atrophicans and 1 lymphocytoma). Artritis was present in 18 cases. Neurological manifestations included 16 cases of meningitis (2 with encephalitis), 11 of craneal neuropathy and 25 of peripheral neuropathy (13 of polyneuropathy). Cardiac manifestations acepted was 2 atrioventricular blockade. We detected 21% of false positive serology and in 33% of the positive cases, the LB criteria was not met. CONCLUSIONS: The low percentage of cutaneous manifestations is probably due to the fact that this series consist predominantly of hospital patients. In Spain, as in the rest Europe the predominant manifestations are neurological, however artritis are not infrequent manifestations. PMID- 8714153 TI - [Tuberculin reactivity in the elderly. Comparison of PPD-RT23 and PPD-CT68]. AB - BACKGROUND: Tuberculin reactivity varies on the basis of the PPD lot and the population studied. AIMS: To compare tuberculin reactivity in the elderly following successive applications of PPD from 2 different lots. POPULATION: Elderly nursing-home residents, from the province of Guadalajara, Spain. METHODS: The elderly were randomly selected and administered with a first Mantoux intradermoreaction of 2 UT of PPD-RT23 or 2 UT of PPD-CT68. The study was performed in a scaled basis, eliminating the reactors (transversal induration > or = 5 mm at 72 h). The non reactors were restudied with a second and third test in alternative arms at a 7-day interval with the same lot and initial dosis of PPD. At one year the scaled study was repeated with 2 UT of PPD-CT68. RESULTS: Eight hundred and thirty elderly persons (79 +/- 9 years) were studied, with 410 receiving PPD-RT23 and 420 receiving PPD-CT68. No differences were observed in the percentage of initial reactors (31% vs 29%; p = 0.26) or in the booster reactors (23% vs 20%; p = 0.16). The PPD-RT23 was found to trigger more intensive reactions than the PPD-CT68 leading to a greater frequency of indurations > 14 mm. After one year, the percentage of stable reactions was similar in both groups (50.5% vs 49.5%; NS). CONCLUSIONS: 1) In elderly nursing-home residents 2 UT of PPD-RT23 and 2 UT of PPD-CT68 showed similar results in regard to the number of reactors and the frequency of booster reactions. 2) The proportion of intense reactions was greater with 2 UT of PPD-RT23 than with 2 UT of PPD-CT68. 3) The stability of the reactions at one year was independent of the lot used in the first phase. PMID- 8714154 TI - [Antibiotic resistance to erythromycin, clindamycin and tetracycline of 573 strains of Streptococcus pyogenes (1992-1994)]. AB - BACKGROUND: The aim of this study was to know the antibiotic resistence of Streptococcus pyogenes to erythromycine, clindamycine and/or tetracycline in community samples. The second aim was to determine the existence of multiresistant strains and to know the relationship between resistant strains, clinical samples and age of the patient. METHOD: A retrospective analysis was performed in all the strains of S. pyogenes isolated from January 1992 to December 1994. Antibiotic sensitivity was studied by MIC by the microdilution method using the Pasco semiautomatic system. RESULTS: During the study period 573 beta hemolytic streptococci were identified as S. pyogenes. The global resistance to erythromycine (2.8%), clindamycine (1.4%) and tetracycline (7.3%) remains at low levels but has significantly increased in the case of erythromycine (p < 0.05) and tetracycline (p < 0.05) over these 3 years. The incidence of strains resistant to clindamycine has also increased slowly although this rise is not significant. Five strains (0.9%) were not sensitive to the three antibiotics studied, 4 being isolated in the last trimester of 1994 in pharyngeal exudates. S. pyogenes resistant to erythromycine was most frequently isolated from cutaneous lesions and in pediatric patients (under the age of 14 years). CONCLUSIONS: These results confirm the trend towards an increase in the number of strains of S. pyogenes resistant to erythromycine, clindamycine and/or tetracycline, being most often found in cutaneous lesions and pediatric patients. PMID- 8714155 TI - [Tuberculin test in HIV infection]. AB - BACKGROUND: Intravenous infected-HIV drug abusers (IDAP/HIV+) are known to have a higher risk of tuberculosis (TB) than others. The effectiveness of tuberculin testing (Mantoux) in the detection of the TB infection in HIV(+) patients is probably lower than HIV(-) individuals. The aim of this paper is to assess tuberculin testing in the active search of cases of TB infection in IDAP individuals and their differences between HIV(+) and HIV(-) ones. METHODS: We studied 332 patients with intravenous drug use belonging to a therapeutic community and to in-patients. Tuberculin testing was performed on all of them. They were examinated if had been previously vaccinated with BCG and we carried out the counting of linfocites CD4 in HIV(+) ones. In the cases of negative tuberculin skin test a second test was administered after 7 days. The patients were grouped in HIV(+) and HIV(-) and the variables were statistically assessed by the chi 2 and a logistic regression model. RESULTS: Tuberculin testing resulted positive in 16.9% IDAP/HIV(+) in comparison with 39.9% of IDAP/HIV(-). The induration size was significantly smaller in HIV(+) than HIV(-) individuals. In both cases, we found that in BCG vaccinated patients positive tuberculin testing was significantly lower than negative tuberculin. The booster effect was detected in 8.9% of HIV(+), whereas in HIV(-) not BCG vaccinated it was of 21.4% and in HIV(-) BCG vaccinated it increased to 30.9%. When the level of the linfocites CD4 exceeds 500/mm3 in HIV(+) the percentage of Mantoux test (+) equals HIV(-) individuals. CONCLUSIONS: A decrease in sensitivity of tuberculin testing in the diagnosis of TB infection in IDAP/HIV(+) was detected. A high percentage of booster effect in IDAP was also demonstrated. We recommend the performance of tuberculin testing at the beginning of HIV infection. PMID- 8714156 TI - [Clostridium difficile and diarrhea associated with the use of antibiotics in the origin of nosocomial and community-acquired diarrhea]. AB - BACKGROUND: Clostridium difficile is currently recognized as an important nosocomial enteric pathogen. The significance as etiologic agent of community and nosocomial diarrhea is not well known in Spain. METHODS: Retrospective study of all cases of community diarrhea that required admission in the hospital and all nosocomial diarrhea observed in a period of three months in a 450-bed university hospital. We performed conventional coprocultives and detection of toxin-A of C. difficile with the method ELISA Premier. RESULTS: During the period of study were included 66 patients, 19 pediatrics and 47 adults patients (23 males, 24 females, age: 54.5 +/- 21.8 years). Three cases (15.8%) of pediatrics patients were diagnosticated of antibiotic-associated diarrhea, only one case were of nosocomial origin. Toxin A of C. difficile were detected in 6 cases, all were patients under two years old, represented 60% of these patients. The origin of diarrhea were: community in 32 cases and nosocomial in 15 of adults patients, in 18 cases (38.3%) were diagnosticated antibiotic-associated diarrhea, 11 were nosocomial. Toxin A of C. difficile were detected in 12 patients, 25.5% of adults, and 4 cases had criteria of C. difficile associated diarrhea, representing 8.5% of the diarrhea. None of this cases were suspected during admission. CONCLUSIONS: Antibiotic-associated diarrhea and C. difficile associated diarrhea were not infrequent cause of diarrhea of nosocomial and of community origin in our environment. We recommended culture and/or detection of toxins of C. difficile in patients who were treated with antibiotics and diarrhea of more than 72 hours of evolution. PMID- 8714157 TI - [Primary cutaneous cryptococcosis and bacterial pneumonia caused by Pseudomonas aeruginosa in AIDS: clinical case and review of the literature]. AB - BACKGROUND: Clinically symptomatic infection due to Cryptococcus neoformans is found in 5-10% of patients with AIDS. It usually appears as meningitis with or without associated blood stream infection. In the last years, severe Pseudomonas aeruginosa infections in HIV (+) patients, with a high morbidity and mortality, are increasingly reported. MATERIALS AND METHODS: A 27 year-old male HIV (+) patient, with previous opportunistic infections, was referred because of a nodular lesion on his left wrist, with an axillar homolateral lymph node of long term evolution. Cryptococcus neoformans var. neoformans was isolated in pure culture from the lesion. Disseminated infection or affection of other organs was ruled out. Therapy with intravenous amphotericin B was initiated, but the patient developed an early nosocomial bacteremic pneumonia due to P. aeruginosa with a fatal outcome. A review of the literature about both complications in AIDS is carried out. RESULTS: The fungal clinical picture is compatible with primary cutaneous cryptococcosis that is a rare condition (12 clinical reports until 1993), of which only one case report in an AIDS patient has been described in 1994. Since 1990 severe infections due to P. aeruginosa in HIV(+) patients are increasingly reported, and pneumonias, bacteremias, catheter-related sepsis, urinary tract infections, intraabdominal infections, central nervous system and othorrhinolaryngologic infections have been described in patients with AIDS in the English-language literature. CONCLUSIONS: Primary cutaneous cryptococcosis is another rare form of infection with C. neoformans in patients with AIDS. The increasingly reported cases of severe infections due to P. aeruginosa could have important implications in the therapy of patients with AIDS. PMID- 8714158 TI - [Rotavirus infection: clinical characteristics and time of elimination of the rotavirus antigen in the feces]. AB - BACKGROUND: A prospective study was performed during three years with 76 children hospitalized with the diagnose of rotaviral gastroenteritis. The aim of our study were to evaluate the clinical features of the rotaviral gastroenteritis, the excretion time of rotavirus in stools and the nosocomial rotavirus infection. METHODS: The detection of the rotaviral antigen in stool was performed using a rapid test based on latex aglutination (Rotalex). For the determination of the excretion time of rotavirus in stools the test was performed daily in 69 children until the results was negative. Furthermore, a clinical study of the patients was carried out. RESULTS: In 39 cases (51.3%) the gastroenteritis was acquired in the community and 37 cases (48.7%) were nosocomially acquired. The most frequent clinical presentation was an acute diarrhea (96%) with vomiting (63.2%) and fever (63.2%). Only three patients did not have diarrhea. The mean duration of diarrhea was between 3-4 days with a highest duration of 10 days. Most of the patients (58%) had an excretion time of rotaviral antigen lower or equal than 3 days, with an extreme value of 14 days. CONCLUSIONS: In our study rotaviral infection occurred mainly in the cooler winter months and in children younger than 6 months of age. A high nosocomial infection level (48.7%) was detected. Vomiting often preceded the onset of diarrhea and fever. The excretion time of rotaviral antigen was independent of the clinical course. PMID- 8714159 TI - [Mechanism of action and resistance to fluoroquinolones]. PMID- 8714160 TI - [Cutaneous lesion and bacteremia of fatal outcome]. PMID- 8714161 TI - [Study of nosocomial infections: incidence or prevalence]. PMID- 8714162 TI - [Prevalence of hospital infections in a regional hospital in Valencia]. PMID- 8714163 TI - [Blood culture systems]. PMID- 8714164 TI - [Isolation of a strain of Campylobacter jejuni resistant to fluoroquinolones and sensitive to nalidixic acid]. PMID- 8714165 TI - [Acute renal insufficiency caused by a sulfadiazine in a patient with cerebral toxoplasmosis and AIDS]. PMID- 8714166 TI - [RIBA 3.0 in positive anti-HCV samples using enzyme immunoassay]. PMID- 8714167 TI - [Septic shock caused by Escherichia coli resistant to a ciprofloxacin after extracorporeal shockwave lithotripsy]. PMID- 8714168 TI - [Rhabdomyolysis and Q fever]. PMID- 8714169 TI - [Clinical experience in switching patients from standard levodopa to a controlled release levodopa]. AB - We switched 50 patients with Parkinson's disease and motor fluctuations from standard levodopa (L-S) to controlled-release carbidopa/levodopa (L-CR). Ten (20%) patients stopped treatment due to adverse events. Data on the 40 patients who completed 6 months of treatment were analyzed. Parkinsonian assessments were performed using the Unified Parkinson's Disease Rating scale (UPDRS) and a nocturnal rating scale. During treatment with L-CR the total daily dose of levodopa increased and the number of daily doses decreased. After changing to L CR, patients also experienced significant improvement on sub-scales II and III of UPDRS and on the Schwab and England and nocturnal mobility scales. The proportion of "off" time decreased but the duration and severity of dyskinesias increased. Most of the patients who dropped out of the L-CR group had already experienced some type of adverse event during treatment with L-S. At the end of the study, the number of patients preferring L-CR was significantly higher than the number preferring L-S. PMID- 8714170 TI - [Drug-induced or aggravated parkinsonism: clinical signs and the changing pattern of implicated drugs]. AB - To determine the prevalence, clinical signs and course of drug-induced Parkinsonism (DIP) in a general neurology practice, as well as to study the changing pattern of drugs implicated. Retrospective study of DIP patients seen between January 1981 and December 1993. Of the 306 cases of parkinsonism seen, 56.8% were induced or aggravated by drugs. This side effects is more frequent in women and often occurs in old age. The drugs implicated most often were cinnarizine, sulpiride and flupentixol. Forty-two patients took 2 drugs simultaneously, whereas 6 took 3. The number of DIP seen increased after 1986 and then remained stable through 1993. Between 1981 and 1988, the drug most often implicated was cinnarizine, though its relative impact decreased in later years. The most frequently seen form of presentation was rigidakinetic syndrome. Neither drug nor age influenced clinical presentation. Parkinsonism disappeared completely within a mean of 5 months in 142 (82%) patients. Twenty-eight (16%) developed Parkinson's disease. Six of them were symptom-free for 12 to 72 months (mean 40 months), whereas 22 never experienced relief from parkinsonism. Sixteen suffered tardive dyskinesia, a complication that was not associated with the use of any particular drug. More than half of the cases of parkinsonism seen in a neurology practice are drug induced or aggravated, generally by psychotropic drugs. The frequency has held steady for the past 6 years. The drugs implicated change as knowledge of their inducement of parkinsonism becomes known. The clinical picture is usually reversible. PMID- 8714171 TI - [The effect of piracetam on involuntary movements in Huntington's disease. A double-blind, placebo-controlled study]. AB - Piracetam and its derivatives, a group of "nootropic" drugs thought to act as metabolic enhancers in the brain, have been widely used to treat a variety of cognitive disorders. Two studies have reported that this compound reduces the severity of involuntary movements in acute onset hemichorea. We therefore randomly assigned 11 patients with Huntington's disease to receive high doses of intravenous piracetam (12 g in 250 ml of saline) or placebo on different days. A blinded observer then rated the severity of involuntary movements over a period of 4 hours. Significant aggravation (p < 0.05) of chorea was seen in patients taking piracetam, in comparison with ratings for both baseline and placebo, 4 hours after administration but not during earlier assessments. Although the mechanism causing this effect is unidentified, we believe that our findings indicate that piracetam should not be used to treat the cognitive disorders accompanying Huntington's disease. PMID- 8714172 TI - [Migraine]. PMID- 8714173 TI - [Functional MRI of the motor area]. PMID- 8714174 TI - [Simulated craniotomy]. PMID- 8714175 TI - [Botulinum A toxins in the treatment of spasticity in cerebral palsy during childhood]. AB - We evaluated the safety and efficacy of Botulinum- A toxin (BTX) in patients with equinus deformity associated with cerebral palsy (CP). We prescribed BTX to six patients (all of whom had previously participated in a multicenter, randomized, double blind study of the same drug); treatment continued for at least 15 months. Four children showed striking improvement, being converted from toe-toe to consistent or occasional heel-toe gait. No side effects were observed. BTX appears to be safe and effective in patients with CP. PMID- 8714176 TI - [Parkinsonism after lethargic encephalitis: possibly the first Spanish case in the second half of the century]. AB - We present the case of a 38-year-old patient who developed symptoms are parkinsonian syndrome at the age of 14, after meningoencephalitis meeting the criteria for a diagnosis of lethargic encephalitis (LE). The extrapyramidal symptoms are asymmetric are quiescent and have responded successfully to levodopa therapy for over 20 years. This is the first such case of parkinsonism secondary to LE to have been described in the Spanish medical literature in recent decades. PMID- 8714177 TI - [Subependymal nodular heterotopia and epilepsy]. PMID- 8714178 TI - [Subependymal nodular heterotopia: morphological variability]. PMID- 8714179 TI - [Riluzole for everybody or for nobody]. PMID- 8714180 TI - [Pneumococcal meningitis and third generations cephalosporins]. PMID- 8714181 TI - [Hemorrhagic colitis caused by verotoxigenic Escherichia coli. Presentation of 9 cases]. AB - OBJECTIVE: To describe the clinical and epidemiological characteristics of nine patients with enteritis caused by verocytotoxin-producing E. coli O157. PATIENTS AND METHODS: Clinical data of patients was collected retrospectively, the isolated strains were tested for verotoxin production (VT) using Vero cell culture line, and presence of VT1 and VT2 gene sequences was detected using amplification techniques (PCR), biotype was also determined using twelve biochemical tests, and genomic macrorestriction profile (PFGE). RESULTS: The patients' age ranged from 11 months to 70 years. The mean duration of diarrhea was 4.7 days. All patients but one had abdominal cramps, seven of nine reported hemorrhagic stools and six had fever. Three patients were affected of haematologycal neoplasia and two of them developed hemolytic-uremic syndrome as a complication. All strains produced VT2 and two of them also produced VT1. Epidemiological link between patients has not been established. Three different biotypes had been distinguished between the nine isolated strains. All but two had different macrorestriction profiles. DISCUSSION: The results obtained showed that clinical manifestations are rather inespecific, including fever (6/9 patients) and there is high association of severe complications. The heterogeneity in PFGE results obtained confirms that the cases are not related. PMID- 8714182 TI - [Necrotizing infections of the soft tissues]. AB - OBJECT: In necrotizing soft tissue infections (NSTI) may be implicated skin, subcutaneous tissue, fascia and skeletal muscle, and include different clinical entities associated with high morbidity and mortality rates. Diagnosis must be early and its initial management must be common. METHODS: A retrospective study of 24 patients diagnosed of NSTI is made. Associated factors with the development of these infections, and its treatment are analysed. It is effectuated an statistical analysis of mortality and related factors. RESULTS: Five cases were Necrotizing Cellulitis, fourteen Necrotizing Fasciitis, three Myonecrosis, one gangrenous pyoderma and one case was a disseminated gluteus abscess. Surgical treatment was resection and debridament--in every case reaching to healthy tissue. Broad-spectrum antibiotics were always associated to surgery. E. coli and Bacteroides have been the most frequent microorganism isolated. 23 patients were operated once at least; 66 surgical interventions were made (mean: 2.86; range: 1 7). Overall mortality of this series was 29%. CONCLUSIONS: Early and aggressive treatment are the corner stone in the management of these patients. With such therapeutic premise, in our series we had a mortality of 29%. Factors associated with a worse outcome have been advanced age, the presence of an underlying disease in the moment of developing the infection and high levels of urea at admittance. PMID- 8714183 TI - [Description of 31 pediatric cases of infection caused by human Parvovirus B 19]. AB - FUNDAMENTALS: [corrected] Human Parvovirus B19 has been identified as the etiological agent implicated in several clinical forms of disease. The aim of this study was to define the clinical aspects observed in the children with human parvovirus infection in our hospital. METHODS: A retrospective study was done in 31 pediatric cases observed in our hospital from January-92 to July-94; all had positive IgM against Parvovirus (determined by enzymoinmunoanalysis). RESULTS: The most frequent signs and symptoms were fever in 66.6% of the cases and skin findings in 48%, manifested as typical infectious erythema in 26% and polimorphic nonspecific exanthema in the rest. There were variable blood manifestations, observed in 11 children (35%): Immune Thrombocytopenic purpura, Chronic anemia in immunocompromised hosts, bone marrow erithrophagocytosis and hemolytic crises in non-hemolytic anemia patients. The abnormal blood findings were mostly observed in children with compromised immunity. Articular disease was presented in four children (12%). Long lasting fever was observed in only one case. CONCLUSIONS: In our sample the diversity of clinical forms of presentation of Parvovirus B19 infection is well documented as it is reported by several authors. Although other studies as polymerase chain reaction (PCR), nucleic acid hybridization, direct visualization of the virus or its particles, or viral antigen detection are desirable to confirm the serologic studies. PMID- 8714184 TI - [Pyomyositis in Zamora]. AB - BACKGROUND: Pyomyositis is a purulent infection involving the skeletal muscle. Although it was initially described in tropical countries, is it ever more frequently found in warm climates. METHODS: A review of the clinical histories of the admission of patients in the Internal and Infectious Medicine Units over the period from May 1992 to April 1994 (1818 admissions) was carried out analyzing those in whom the diagnosis was of pyomyositis. This diagnosis was performed by clinical data and radiologic and microbiologic confirmation. RESULTS: Five patients diagnosed with pyomyositis were found. Of the five cases, four were men with ages ranging from 27 to 64 years. In most of the cases more than one muscle group was involved and only two cases showed predisposing factors (history of injury and DM). The existence of abscesses was shown by echography in 3 cases and by CAT in 2 cases. Staphylococcus aureus was the most frequently isolated microorganism. The five patients underwent medical treatment and surgical treatment was also performed in 2 achieving satisfactory evolution. CONCLUSIONS: Imaging techniques (echography and CAT) effectively contribute to the diagnosis of pyomyositis. Medical treatment, associated with surgery in some cases, achieved satisfactory evolution. The incidence of pyomyositis is probably greater than what has been recognized to date. PMID- 8714185 TI - [Detection of resistance to the amoxicillin-clavulamic acid combination in Salmonella and Shigella]. AB - PURPOSE: To study the emergence of amoxicilline/clavulanate resistance in Salmonella and Shigella. MATERIAL AND METHODS: We have studied 118 strains of Salmonella enterica and 11 strains of Shigella flexneri isolated in our laboratory along 1993. The MICs values for amoxicilline/clavulanate were determined by the E-Test method and confirmed by the standard agar dilution method. RESULTS: We have observed that 6% (7 out of 118 strains) of Salmonella enterica and 42% (5 out of 11 strains) of Shigella flexneri isolates exhibited an intermediate susceptibility or were resistant to amoxicilline/clavulanate with MICs values ranging between 16 mg/l and 32 mg/l. CONCLUSIONS: Along 1993 we have detected the emergence of an amoxicilline/clavulanate resistance in salmonellae and shigellae strains. To our knowledge this fact has not been published up to now (Medline, 1990-October 1994). We don't know the clinical importance of this fact. PMID- 8714186 TI - [Neurosurgical infections]. PMID- 8714187 TI - [Current status of antifungal prophylaxis in opportunistic mycoses]. PMID- 8714189 TI - [Cefprozil]. PMID- 8714188 TI - [Tuberous lesion in the dorsum of the hand in an immunocompetent patient]. PMID- 8714190 TI - [What do we know about hantavirus?]. PMID- 8714191 TI - [Disseminated histoplasmosis in a patient infected with the HIV]. PMID- 8714192 TI - [Bacteremia caused by Campylobacter jejuni associated with spontaneous abortion]. PMID- 8714193 TI - [Bacteremia and spontaneous peritonitis caused by serogroup C Neisseria meningitidis in a cirrhotic patient]. PMID- 8714194 TI - [Coxofemoral infectious arthritis caused by Candida albicans in an HIV seropositive patient]. PMID- 8714195 TI - [Pneumocystis carinii pneumonia. Diagnosis using immunofluorescence with monoclonal antibodies]. PMID- 8714196 TI - [Nocardia asteroides pneumonia in a patient undergoing a kidney transplant]. PMID- 8714197 TI - [Intrafamily infection caused by Parvovirus B19: different manifestations according to age]. PMID- 8714198 TI - How and why ribosome? PMID- 8714199 TI - Time-resolved emission spectroscopy as a tool to follow nucleic acid-protein interaction. AB - Fluorescence spectroscopy is undoubtedly a useful tool to study the structural and functional aspects of nucleic acids-protein interactions as well as the catalytic functions of particular residues of multi-subunit enzyme complexes. The dynamic interaction of nucleic acids and proteins occurring at nanosecond time scale can now be monitored by making life-time measurements or by time-resolved emission spectroscopy. These measurements are made by exploiting the intrinsic fluorescent residues in proteins i.e. W or by the use of extrinsic fluorophores which are tagged on to particular residues and that are sensitive to the microenvironment changes. In this study we describe the use of time resolved emission spectroscopy to (a) analyse the transient binding between sigma 70 and DNA by monitoring the quenching of W residues and (b) monitor the various states which nucleosomes of active, inducible or inactive chromatin may adopt in vivo. PMID- 8714200 TI - Molecular analysis of the promoter region of the gene encoding the beta-amyloid precursor protein. AB - The amyloid beta-peptide (approximately 4 kDa-M(r)) is generated by the proteolytic cleavage of a larger beta-amyloid precursor protein (beta APP) encoded by a gene on chromosome 21. The abnormality in gene regulation of beta APP may be an important factor in the neuropathology of Alzheimer's disease. The control of transcription is mediated by different DNA regulatory elements (cis acting) present in the promoter of the gene. There are about 26 DNA motifs, present in the immediate 5'-flanking region of the beta APP gene, through which various cell/tissue-specific factors (trans-acting) can exert their influence on transcription. Here, the effects of nerve growth factor (NGF), basic fibroblast growth factor (bFGF), phorbol 12-myristate 13-acetate (PMA), interleukin-1 (IL-1) and retinoic acid (RA) on promoter activity of the beta APP gene were analyzed. To investigate the effect of these factors on promoter activity, a recombinant plasmid which contained sequences of -489 base pairs (bp) from the 5'-flanking region of the beta APP gene was used. The truncated region of the promoter was linked upstream to a reporter gene, chloramphenicol acetyl transferase (CAT). Promoter activity was tested by transient transfection of the fusion plasmid in PC12 cells using the electroporation method (960 microF at 350 V). The treatment of PC12 cells with either NGF, bFGF, PMA, IL-1 or RA stimulated the activity of the beta APP promoter. Treatment of cells with either NGF or bFGF resulted in a higher level of stimulation in the basal level of promoter activity than when cells were treated with either PMA, IL-1 or RA. The pre-treatment of cells with these factors for a duration of 4 days prior to transfection with the promoter plasmid was necessary for the stimulatory effect. The cells that were treated with either of these factors after transfection showed no significant change in the basal level of promoter activity. Thus, certain growth factors and a cytokine could enhance the basal level of promoter activity of the beta APP gene, suggesting a possible participation of a growth-factor (s)-mediated transcriptional element in the control of gene expression of beta APP. PMID- 8714201 TI - Interaction of yeast elongation factor 3 with polynucleotides, ribosomal RNA and ribosomal subunits. AB - In addition to the two usual eukaryotic elongation factors (EF-1 alpha and EF-2) fungal ribosomes need a third protein, elongation factor 3, for translation. EF-3 is essential for in vivo and in vitro protein synthesis. Functionally, EF-3 stimulates EF-1 alpha dependent binding of aminoacyl-tRNA to the ribosomal A site when E site is occupied by deacylated tRNA. EF-3 has intrinsic ATPase activity which is regulated by the functional state of the ribosome. EF-3 ATPase is activated by both 40S and 60S ribosomal subunits. However intact 80S ribosomes are needed for efficient activation of EF-3 ATPase. EF-3 appears to be an RNA binding protein with high affinity for polynucleotides containing guanosine rich sequences. To determine whether guanosine rich sequence of ribosomal RNA is involved in EF-3 binding, an antisense oligonucleotide dC6 was used to block EF-3 interaction with the ribosome. The oligonucleotide suppresses activation of EF-3 ATPase by 40S ribosomal subunit and not by the 60S or the 80S particles. Poly(U) directed polyphenylalanine synthesis by yeast ribosomes is inhibited by dC6. To define the binding site of the oligonucleotide and presumably of EF-3 on 18S ribosomal RNA, hydrolysis of rRNA by RNase H was followed in the presence of dC6. These experiments reveal an RNase H cleavage site at 1094GGGGGG1099 sequence of 18S ribosomal RNA. This guanosine rich sequence of rRNA is suggested to be involved in EF-3 binding to yeast ribosome. Data presented in this communication suggest that the activity of EF-3 involved a direct interaction with the guanosine rich sequence of rRNA. PMID- 8714202 TI - Identification of the elongation factor Tu binding site on 70S E. coli ribosomes by chemical crosslinking. AB - Elongation factor Tu (EF-Tu), in the presence of Phe-tRNA, GMPPCP, and Poly (U), binds to 70S ribosomes at the recognition (R) site. In order to identify the ribosomal proteins adjacent to the EF-Tu occupying the R site, EF-Tu:Phe tRNA:GMPPCP:ribosome complexes were crosslinked by modification with 2 iminothiolane and mild oxidation to form disulfide bridges between neighbouring proteins whose endogenous or introduced SH groups were appropriately located. The binding of Phe-tRNA to the ribosome was shown to be largely dependent on the presence of Poly(U). The total protein from the complexes was extracted and separated by two-dimensional gel electrophoresis by non-equilibrium pH gradient electrophoresis (NEpHGE) in the first dimension, followed by gradient SDS gel electrophoresis in the second dimension. Comparison of control samples crosslinked without Poly(U) to those crosslinked with Poly(U) present showed a single crosslinked complex in the region of the gel near EF-Tu. No cross-links in the vicinity of EF-Tu were visible in the absence of Poly(U). The crosslinked proteins in this region were recovered by electroelution, radiolabeled and their identity was confirmed by 2D gel electrophoresis and immunoblot analyses. Two major 50S ribosomal proteins, L7/L12 and L10 were found to be covalently linked to EF-Tu. The isolated crosslinked complex did not contain any protein from the 30S subunit. These results demonstrate that L7/L12 and L10 are the major, if not only, ribosomal protein cross-links to EF-Tu in the R site. In contrast to previous crosslinking results obtained by others, our results define a unique location for the EF-Tu binding site, one compatible with functional data and near that of the EF-G binding site on the ribosome. PMID- 8714203 TI - Tat mediates transcriptional activation of HIV-1 gene in vitro. AB - Gene expression of human immunodeficiency virus (HIV-1) is greatly enhanced by a viral transactivator, the Tat protein, which interacts with R region sequences of the HIV-1 long terminal repeat (LTR). There is no direct evidence to indicate transcriptional activation of HIV-1 by Tat. Using an in vitro transcription system, we demonstrate that an established mouse cell line, which constitutively expresses Tat protein, selectively stimulates the steady state levels of the transcripts directed from the long terminal repeat (LTR) sequences of HIV-1. The gel binding retardation assays further demonstrate a stable activated complex, formed due to direct binding of Tat to DNA elements of the HIV-1 LTR. These data implicate transcription as the site of Tat action in trans-activation and could play an essential role in human immunodeficiency virus replication, similar to the nuclear trans-activators of other viruses. PMID- 8714204 TI - Expression of the Japanese encephalitis virus NS3 and NS2b proteins as glutathione S-transferase fusions. AB - Flaviviruses generate their structural and nonstructural proteins by proteolytic processing of a single large polyprotein precursor. These proteolytic events are brought about both by host cell signalase and a virally encoded protease. The virally encoded proteolytic activity has been shown to reside within the nonstructural protein 3 (NS3) and requires the product of the nonstructural 2b (NS2b) gene. In order to obtain sufficient quantities of pure NS2b and NS3 proteins for kinetic analysis, we have expressed both these proteins in recombinant systems as fusions to glutathione S-transferase (GST). The fusion constructs were driven by the strong bacteriophage T7 promoter. Transfection of these constructs into the African green monkey kidney cell line CV-1 previously infected with a recombinant vaccinia virus expressing the T7 RNA polymerase resulted in synthesis of the fusion proteins. Both the fusion proteins could be purified to homogeneity in a single step using a glutathione agarose affinity matrix. PMID- 8714205 TI - Cloning and characterization of mycobacteriophage I3 promoters. AB - Restriction fragments of mycobacteriophage I3 DNA capable of initiating transcription have been cloned into a promoter selection vector of Escherichia coli, and selected on the basis of development of resistance to chloramphenicol. The growth pattern of these 'promoter clones' on a concentration gradient of chloramphenicol and the biochemical assays of the chloramphenicol acetyl transferase have permitted the assessment of their relative promoter strengths. DNA sequence analysis revealed significant homology of these promoters to the -35 regions of the mycobacterial--and E. coli promoter consensus, but less so the -10 region. Based on the sequence of phage I3 promoters identified here and the reported sequences of mycobacterial promoters, a promoter consensus for mycobacteria has been generated. PMID- 8714206 TI - Exclusion of temperature bacteriophage P22 by virulent bacteriophage MB78 of Salmonella typhimurium: competition for association with the replication complex. AB - MB78, a virulent bacteriophage of S. typhimurium does not allow other bacteriophages like P22 and 9NA to multiply in its presence. The exclusion of P22 by MB78 is found to be due to competition for common binding site(s) in the host cell membrane. As a result, P22 DNA fails to replicate in presence of MB78 DNA. Further, the sedimentation profile of P22 DNA in cells infected simultaneously with P22 and MB78 suggested fragmentation of P22 DNA. This may also contribute to the exclusion phenomenon. PMID- 8714207 TI - Designing of peptides with immuno-modulatory properties using protein A as a probe. AB - A series of reports from our laboratory have described the multifarious properties of protein A of Staphylococcus aureus Cowan I, apart from its IgG binding affinity. Original reports regarding its anti-tumor, anti-toxic, anti carcinogenic and immunomodulatory properties published earlier by the authors have implicated some uniqueness of this bacterial protein. It was conceived that such diversified properties must lie in its specific peptide sequences, rendering it to act and behave as a multipotent "Biological Response Modifier" (BRM). The high resolution X-ray structure of protein A-Fc complex has been delineated earlier, and has been the foundation of many protein engineering studies. This structure along with the amino acid sequence data of its four repetitive domains provided us the basis for designing an octapeptide. This octapeptide was synthesized by solid phase peptide synthesis considering it as the probable site through which PA binds IgG. This octapeptide (NH2-Gln-Asn-Ala-Phe-Tyr-Glu-Ile-Leu COOH) is present in the first helical segment of B-domain of protein A, and also is a part of domain D, A and C. This octapeptide has been shown to bind IgG by the immunoblotting technique. The binding affinity of the octapeptide appears to be significantly higher than that of intact protein A, as was revealed by calculation of Ka (association constant) and Kd (dissociation constant) values. This octapeptide might serve as a good immunoadsorbant for IgG and/or immune complexes. PMID- 8714208 TI - Deduced amino acid sequence of 2S storage protein from Brassica species and their conserved structural features. AB - 2S seed storage albumin coding regions from five Brassica species, namely Brassica campestris, B. oleracea, B. nigra, B. juncea, and B. carinata have been cloned by PCR amplification of genomic DNA using oligonucleotide primers and their nucleotide sequences have been determined. These sequences showed more than 85% homology amongst themselves and considerable homology with some other crucifer 2S protein coding sequences. The deduced amino acid sequences showed more homology due to some inconsequential mutations in codons without changing the amino acids. Computer analysis of the protein sequences for possible secondary structure revealed a high degree of conservation of hydrophilic and hydrophobic domains and the invariant positions of cysteine residues. Unrooted phylogenic tree based on the coding region of 2S albumin from different Brassica species cloned by us and published sequences from other Cruciferae indicated that these genes originated before the evolutionary divergence of different Brassica species and were conserved due to some stringent structural and functional features required for seed metabolism. PMID- 8714209 TI - Intra- and interloop interactions in the folded G quartet structure of Oxytricha telomeric sequence. AB - The antiparallel intramolecular G quartet structure for the 3.5 copy Oxytricha telomeric sequence d(G4T4)3G4 has been established using a combination of spectroscopic and chemical probing methods. In the presence of Na+ ions, this sequence exhibits a circular dichroism spectrum with a positive band at 295 nm and a negative band around 265 nm, characteristic of an antiparallel G quartet structure. Further, we show that d(G4T4)3G4 adopts an antiparallel intramolecular G quartet structure even in K+ unlike d(G4T4G4). KMnO4 probing experiments indicated the existence of intra and interloop interactions in the Na+ induced structure. We have found that K+ not only increases the thermal stability of G quartet structure but also binds to the loop region and disrupts stacking and interloop interactions. Biological consequences of such cation-dependent conformational micro-heterogeneity in the loop region of G quartet structures is also discussed. PMID- 8714210 TI - Molecular mechanics studies of homopolymeric and mixed sequence Py.Pu*Pu triple helices. AB - DNA triple helices containing two purine strands and one pyrimidine strand (C.G*G and T.A*A) have been studied, using model building followed by energy minimisation, for different orientations of the third strand resulting from variation in the hydrogen bonding between the Watson-Crick duplex and the third strand and the glycosidic torsion angle in the third strand. Our results show that in the C.G*G case the structure with a parallel orientation of the third strand, resulting from Hoogsteen hydrogen bonds between the third strand and the Watson-Crick duplex, is energetically the most favourable while in the T.A*A case the antiparallel orientation of the third strand, resulting from reverse Hoogsteen hydrogen bonds, is energetically the most favourable. These studies when extended to the mixed sequence triplexes, in which the second strand is a mixture of G and A, correspondingly the third strand is a mixture of G and A/T, show that though the parallel orientation is still energetically more favourable, the antiparallel orientation becomes energetically comparable with an increasing number of thymines in the third strand. Structurally, for the mixed triplexes containing G and T in the third strand, it is seen that the basepair non isomorphism between the C.G*G and the T.A*T triplets can be overcome with some changes in the base pair parameters without much distortion of either the backbone or the hydrogen bonds. PMID- 8714211 TI - The precision of regulation in Dictyostelium discoideum: implications for cell type proportioning in the absence of spatial pattern. AB - We have made careful counts of the exact number of spore, stalk and basal disc cells in small fruiting bodies of Dictyostelium discoideum (undifferentiated amoebae are found only rarely and on average their fraction is 4.96 x 10(-4)). (i) Within aggregates of a given size, the relative apportioning of amoebae to the main cell types occurs with a remarkable degree of precision. In most cases the coefficient of variation (c.v.) in the mean fraction of cells that form spores is within 4.86%. The contribution of stalk and basal disc cells is highly variable when considered separately (c.v.'s upto 25% and 100%, respectively), but markedly less so when considered together. Calculations based on theoretical models indicate that purely cell-autonomous specification of cell fate cannot account for the observed accuracy of proportioning. Cell-autonomous determination to a prestalk or prespore condition followed by cell type interconversion, and stabilised by feedbacks, suffices to explain the measured accuracy. (ii) The fraction of amoebae that differentiates into spores increases monotonically with the total number of cells. This fraction rises from an average of 73.6% for total cell numbers below 30 and reaches 86.0% for cell numbers between 170 and 200 (it remains steady thereafter at around 86%). Correspondingly, the fraction of amoebae differentiating into stalk or basal disc decreases with total size. These trends are in accordance with evolutionary expectations and imply that a mechanism for sensing the overall size of the aggregate also plays an essential role in the determination of cell-type proportions. PMID- 8714212 TI - Contextual constraints in the choice of synonymous codons. AB - From EMBL Nucleotide Sequence Database, protein coding sequences of all E. coli and its DNA phages, were extracted using our computer programme. Same programme has been used to form a database of sequence of oligonucleotides of length 18 nucleotides on both sides of each of the 61 codons. From analysis of this database and study of variations in twist parameter (Tw) values, as an indicator of sequence dependent variations in B-DNA helix, a method is developed to fix the codon among the set of synonymous codons. The accuracy of the method was checked on enlarged data set by adding data from more prokaryotes. Our method assign the codon 85-90% times correctly if the selection has to be made between codons having different sequence in terms of R and Y. The accuracy of the method is somewhat lower when choice of the codon has to be made between codons having same codes in terms of R and Y. This study points out that the major factors which decide the choice of a codon from a set of synonymous codons are contextual constraints arising from flanking regions. PMID- 8714213 TI - Codon usage in the prototype baculovirus--Autographa californica nuclear polyhedrosis virus. AB - The overall codon usage profile of Autographa californica nuclear polyhedrosis virus (AcNPV) was analyzed, using UWGCG sequence analysis software package from the known protein coding gene sequences available in GenBank Release 72. The analysis revealed that although only 45% of the codon used by AcNPV have G/C at wobble base position, 15 out of 20 AcNPV codons over-utilized for their given amino acids has G/C at the wobble position indicating a possible selection of these codons. The differences in codon usage profile were studied using a parameter called D squared value, calculated with the aid of CORRESPOND program of UWGCG software package. While most of the codon usage profile of the individual genes was very similar to the overall AcNPV codon usage profile (D squared value less than 1.5), there were notable differences (D-squared value greater than 1.5). These genes were polh, p10, ub, sod, gp41, core, 25k, 39k, ie n, etm, ets most of which, interestingly, belonged to late or very late class and were expressed relatively more efficiently. The two highly expressed genes of AcNPV, polh and the p10, differ from the overall AcNPV codon usage profile with respect to at least nine amino acids (Val, Ala, Ser, Lys, Ile, Thr, Leu, Phe, Arg). Our findings that the two highly expressed late genes polh and p10 utilize a codon usage profile different from the early genes have important implications. PMID- 8714214 TI - Utility of a Mycobacterium tuberculosis GC-rich repetitive sequence in the diagnosis of tuberculous pleural effusion by PCR. AB - A GC-rich repetitive sequence (GCRS) of Mycobacterium tuberculosis was identified in our laboratory which displayed a high homology with GC-rich sequences of M. tuberculosis and M. bovis. A PCR assay based on the amplification of the proximal 150 bp of GCRS and its detection by non-radioactive hybridization was developed. The accuracy of the GCRS-based PCR assay was evaluated in a clinical setting for the detection of mycobacterial DNA in pleural fluids for the diagnosis of tuberculosis (TB) using clinical criteria and pleural biopsy histology as gold standard. In a blind study, a total of 67 pleural fluid samples (38 tuberculous and 29 nontuberculous) were analysed by PCR and the results were compared with pleural biopsy, Ziehl-Neelsen staining and culture. Mycobacteria could not be detected by either smear or culture techniques in any of the pleural fluids samples. Out of 38 tuberculous pleural effusions, 24 were positive by PCR (63.2% sensitivity). When PCR results were compared with pleural biopsy histology, an increased sensitivity of 73.3% was obtained. Out of the 29 nontuberculous pleural effusions, 2 false positive results were obtained accounting for an overall specificity of 93.1%. The GCRS-based PCR assay thus has a definite role in the diagnosis of tuberculous pleural effusion in contrast to smear and/or culture techniques. PMID- 8714216 TI - A study of aggregation of 9-(dicyano-vinyl)julolidine. AB - Di(cyano vinyl)julolidine (DCVJ) is a fluorescent probe which has been used to monitor the local mobility of its binding sites on proteins. It shows a concentration dependence of its emission spectrum in water. At higher DCVJ concentrations, a longer wavelength band appears. The latter increases relative to the shorter wavelength band as a function of increased DCVJ concentration. Absorption and excitation spectra indicate that the concentration dependent emission in the longer wavelength is a consequence of association in the ground state and subsequent excimer formation. DCVJ forms two types of complexes with gamma-cyclodextrin, one of which shows the longer wavelength emission band. Analysis of stoichiometry of association also suggests that longer wavelength emission band may be a consequence of association of two molecules of DCVJ in the gamma-cyclodextrin cavity. Possible uses of such excimer formation in biological systems have also been discussed. PMID- 8714215 TI - Kinetoplast DNA minicircle binding proteins in a Leishmania Spp: interference of protein DNA interaction by berenil. AB - A kinetoplast DNA minicircle of a Leishmania Spp. binds to several proteins of the kinetoplast Lysates of kinetoplasts of Leishmania grown in the presence of berenil show complete disappearance of some of these protein bands, while the rest of the proteins present appear as much less intense bands in South Western blots when probed with either the conserved or variable regions of the minicircle or whole minicircle DNA. The conserved region of minicircle DNA complexed with berenil in vitro also fails to interact with the DNA binding proteins of kinetoplast of untreated cell in South Western blots. Since berenil induces dyskinetoplasty of kinetoplastidae, the results indicate that interference of protein-DNA interaction in the presence of berenil may be the primary event in making organisms dyskinetoplastic. PMID- 8714217 TI - Nucleic acid binding properties of a helix stabilising nucleoid protein from the thermoacidophilic archaeon Sulfolobus acidocaldarius that condenses DNA into compact structures. AB - Helix stabilising nucleoid protein (HSNP-C') from an acidothermophilic archaeon Sulfolobus acidocaldarius has been characterised with respect to interaction with nucleic acids by gel retardation assay, binding to nucleic acid columns, fluorescence titrations and electron microscopy. The protein exists in solution as very large multimeric aggregates as indicated by cross-linking studies. The protein binds strongly and co-operatively to double stranded DNA. Electron microscopy of the complexes of the protein with DNA shows compact structures suggesting that the protein condenses DNA. PMID- 8714218 TI - Properties of oligonucleosomes from active genes of rat liver. AB - Active chromatin fraction from rat liver nuclei has been isolated under the lower ionic strength conditions, to prevent salt induced rearrangement and exchange of linker histone H1. Salt induced higher order folding in these oligonucleosomes was determined by sedimentation, viscosity, aggregation and circular dichroism studies. Sedimentation studies indicate that upon raising the ionic strength from 25 mM to 65 mM (mainly NaCl), active oligonucleosomes show intrafragmental interaction and formation of soluble oligomers. These oligomers disaggregate into unfolded monomers at 90 mM ionic strength. In contrast, oligonucleosomes from inactive genes show gradual increase in intrafragmental higher order folding without any interfragmental interaction on raising salt concentration. A much higher decrease in viscosity of active oligonucleosomes in comparison to bulk oligonucleosomes also support the above conclusion. However, on raising salt concentration above 100 mM NaCl, both the chromatin fractions are capable of forming insoluble aggregates. Decrease in the molar ellipticities of bulk oligonucleosomes at 273 and 282 nm is observed on raising ionic strength from 25 mM to 65 mM. A different pattern of this decrease is observed in case of active oligonucleosomes, indicating adaptation of a different type of salt induced secondary structure of DNA in these oligonucleosomes. Melting profile of DNA from active and bulk chromatin suggests that the base composition of both the chromatin fractions is same. PMID- 8714219 TI - Risks from occupational and dietary exposure to mevinphos. AB - Mevinphos (trade name, Phosdrin), a category 1 organophosphorus insecticide, has been used mainly as a cleanup pesticide for vegetable crops. A risk assessment for occupational and dietary exposure to mevinphos was initiated because of the high acute toxicity of the compound. Repetitive dosing with mevinphos did not cause any discernible histopathological effects in mice or rats, nor was it oncogenic in either species. The principal toxic effects of mevinphos, both short and long term, were due to inhibition of cholinesterase activity. Consequently, potential adverse effects from short-term exposures were the primary concern. A human no-observed-effect level (0.025 mg/kg) for cholinergic signs was used as the regulatory basis for calculating margins of safety (MOSs) for potential acute dietary and short-term occupational exposures. Estimates of exposure to mixer/loaders, pilots, and flaggers associated with aerial application of mevinphos were based on passive dosimetry. Because no acceptable exposure studies for work tasks associated with ground application of mevinphos were available, surrogate data based on ground application of oxydemeton-methyl were used. Exposure estimates for field workers and harvesters relied on measured dislodgeable foliar residues of mevinphos and transfer factors generated from studies of other active ingredients. MOSs for mean acute occupational exposure of mixer/loader/applicators associated with ground application and of harvesters working in fruit trees were less than the value conventionally recommended to protect people from the toxic effects of mevinphos. MOSs for the 95th percentile of short-term worker exposure for all mixer/loader work categories associated with mevinphos application were also inadequate. Calculated MOSs for potential acute dietary exposure to measured residue levels of mevinphos were adequate for the various population subgroups. However, 25 of the USEPA tolerances for mevinphos on agricultural commodities were not adequate to protect for the toxic effects of mevinphos from theoretical acute dietary exposure to one or more population subgroups if commodities are consumed with residues at the tolerance level. When the mean short-term occupational exposures were combined with potential acute dietary exposure, the MOSs for mixer/loaders engaged in aerial applications, as well as ground applications, were inadequate to protect people from the toxic effects of mevinphos. As mitigation of the estimated excessive occupational exposures did not appear possible, both California and the USEPA were preparing to cancel registration of the product. However, an agreement was worked out between the manufacturer and the two agencies that ended production for domestic use but allowed existing stocks in the channels of trade to continue to be used for a limited period. PMID- 8714220 TI - Chlorophenols in the terrestrial environment. AB - Chlorophenols are presently widespread in the environment. Even in the most remote natural environments, the presence of chlorophenols in both aquatic and terrestrial food chains has been recorded. These pervasive compounds have been used for a wide range of domestic, agricultural, and industrial purposes for more than 50 years. In addition to industrial production and usage, chlorophenols are produced from naturally occurring phenols as a result of chlorine bleaching of wood pulp in the paper industry and through the chlorination of domestic water supplies and swimming pools. It must be emphasized that chlorophenols, although a local problem in some areas, generally cannot be considered a major environmental problem today because their use is prohibited or restricted in many countries. Future reduction in the use of the herbicidal phenoxy acids will further minimize their levels in the terrestrial environment. Because of the long persistence and high toxicity of pentachlorophenol in particular, however, it is important that the discharge of chlorophenols to the terrestrial environment by way of sewage sludge or pulp mill effluents be maintained at their current levels or even reduced to lower levels. PMID- 8714221 TI - Uranium in the environment: occurrence, transfer, and biological effects. AB - The assessment of uranium chemical and radiological consequences depends on the physicochemical properties of these radioelements and the knowledge of their environmental fate. Although uranium is the source of all these fissionable isotopes, its fate in ecosystems has been poorly investigated. In this review, we have updated information concerning the fate of uranium in the different compartments of the environment, the possibility of transfer to man through the food chain, and the biological and toxicological effects of this metal at cellular, tissular, or organism levels. The physicochemical characteristics of uranium, as well as its regulatory statutes, were reviewed. The fate of uranium in the environment was presented by indicating sources of uranium emission and the possible routes of transfer to man. The biological alterations caused by uranium exposure were discussed, and finally, we presented results collected during our recent study. Some propositions on research to be done to advance the understanding of uranium occurrence in the environment were also given. PMID- 8714222 TI - Epidemiological analysis of persistent organochlorine contaminants in cetaceans. AB - Information is provided to test the hypothesis that organochlorines introduced into the environment since the early 1940s could threaten the reproductive potential of baleen whales and other cetaceans. Comparisons are made using data on the role of organochlorines in a model system, the Great Lakes region of North America, and in model animals, including humans, pinnipeds, and other wildlife. DDT and PCB are used as model organochlorines with the caveat that there may be thousands of other chemicals in the environment also involved. Improved sensitivity in analytical quantification of synthetic chemicals in biological tissue has been accompanied by an increase in knowledge about biochemical processes that control development and function. The effects described in this review are the result of disrupted gene expression, not damage to the gene. The mechanisms of action of the organochlorines reveal their ability to affect developing organisms at very low concentrations during critical life stages: embryonic, fetal, and early postnatal. Exposure during early development can disrupt the organization of the endocrine, reproductive, immune and nervous systems, effecting irreversible damage that may not be expressed until the individuals reach adulthood. The recent discovery that human sperm count is declining worldwide at a rate of 1 x 10(6) sperm/(mL.yr) suggests common exposure to estrogen-like chemicals during prenatal and early postnatal development. This raises concern for other top predator species that also share the same exposure. Periods of intense feeding followed by long periods of fasting are common among species of baleen whales. This unique strategy places the embryonic and nursing calves in vulnerable positions, because under both situations maternal blood levels are elevated as a result of absorption from food intake or as a result of mobilization as fat is metabolized. Estimates of Toxic Equivalents (TEQs) based on the occurrence of four PCB congeners (118, 183, 153, 180) in sigma PCB reported in whales are highest for St. Lawrence belugas and Faroe Island long finned pilot whales. This conservative approach reveals that some whale species are within the range of enzyme-induced TEQs at which effects have been associated with adverse health effects in other aquatic species. The epidemiological approach was used for analysis because it was developed to handle multiple exposure scenarios in which direct causal links are virtually impossible to isolate. The analysis includes the tenets of timeorder, strength of association, specificity of cause and effect, consistency, coherence, and predictive performance. PMID- 8714223 TI - Surgical technique, curability and postoperative quality of life in an extended pancreatectomy for adenocarcinoma of the pancreas. AB - BACKGROUND/AIMS: In the treatment of adenocarcinoma of the pancreas, we have succeeded in both decreasing the locoregional recurrence and improving the patient's survival by adding a wide range of lymphatic and connective tissue clearance to the conventional pancreatectomy. The present paper shows the detailed techniques of this procedure and the factors associated with long-term survival to provide a better understanding of the indications and limitations. MATERIALS AND METHODS: The postoperative survival rate in association with the background factors of the tumor and the quality of life is analyzed in 62 patients who received an extended pancreatectomy of ductal carcinoma of the pancreas. In this procedure, a wide range of lymphatic and connective tissue clearance was cleared in addition to the pancreatectomy, skeletonizing the major vessels. RESULTS: The long-term survivors had the following background factors: tumor diameter 4 cm or less; no nodal involvement or involvement limited to the immediate region of the tumor (n1-group); PV/SMV-invasion was absent or slight (2 cm or less in length and hemilateral). Long-term survival was rarely expected for any patients who had more advanced factors. Since removing the nerve plexi and connective tissues skeletonize major vessels, most patients did not recover their health easily within one post-operative year. CONCLUSION: While our extended pancreatectomy is the most promising procedure at present, providing the candidate is appropriate, its indication should be carefully selected because of the patient's quality of life and cost-performance. PMID- 8714225 TI - Brachytherapy for pancreatic cancer: a review. AB - Brachytherapy has been used for pancreatic cancer for over eight decades. Appositional, interstitial and intraluminal applications have been employed. Data from approximately three hundred cases are summarized in this review. The most common form of brachytherapy. Although this procedure, when combined with external beam radiation therapy and systemic chemotherapy, provides the best possible local control of pancreatic cancer, it is a hazardous procedure in most hands. Hopefully, the new techniques, such as intraluminal brachytherapy and infusional brachytherapy, will less prone to serious complications and they will prove to be efficacious in the management of cancer of the pancreas. PMID- 8714224 TI - Intraoperative radiotherapy and hyperthermia for unresectable pancreatic cancer. AB - Intraoperative radiotherapy (IORT) is a safe and efficient method of delivering a high single dose of radiation to a target. IORT has a significant effect on abdominal pain and back pain in patients with unresectable pancreatic cancer, but IORT alone is not so effective in improving the prognosis. The combination of IORT with external beam radiotherapy (EBRT) can achieve a higher local control rate than EBRT alone, although most previous studies have not necessarily indicated any improvement in survival. We recently found that high-dose IORT (30 Gy) + EBRT (40-60 Gy) was more effective than EBRT alone in patients with a CA 19 9 level < 1000 U/ml. Thus, IORT plus EBRT can be recommended for unresectable but localized pancreatic cancer associated with low tumor marker levels. Hyperthermia combined with radiation or chemotherapy is another possible treatment strategy, but no effective method of delivering external regional hyperthermia has yet been established. Thermometry is another problem with external hyperthermia. At present, intraoperative hyperthermia given in combination with IORT seems to be the best method of heating a pancreatic tumor adequately, but its efficacy is still unclear. PMID- 8714226 TI - A new method of intra-arterial regional chemotherapy with more selective drug delivery for locally advanced pancreatic cancer. AB - BACKGROUND/AIMS: In order to deliver the anti-cancer drugs more selectively into the cancer tissues, we have developed a new method of intra-arterial regional chemotherapy for locally unresectable cancer of the exocrine pancreas. MATERIALS AND METHODS: This method involved placing the catheters selectively into the splenic artery and/or into the gastroduodenal artery during laparotomy. Postoperatively, via the catheter, we infused 50-100 mg of Methotrexate mixed with 10 micrograms of Angiotensin-II with an intent of increasing the blood flow in the tumor tissue but decreasing that to the non-tumor tissues. Simultaneously, a bolus iv-infusion of 5-fluorouracil (5-Fu, 500 mg) was performed. One day after each chemotherapy, citrovorum factor (Leucovorin, 30mg) was given per orally, For 15 patients with locally non-resectable pancreatic cancer, this treatment was repeated weekly or biweekly at our out-patient clinic. RESULTS: As a result, the toxicity was so slight that all patients could tolerate this treatment as long as the catheter was patent (11 +/- 8 postoperative months). The survival period was 16 +/- 9 months (range: 5-36 months; median: 14 months), and one-, two- and three year survival rates were 60%, 23% and 11%, respectively. Patients could take care of themselves within 12 +/- 9 months, and either complete (50%) or partial (50%) pain-relief was obtained among the 12 patients with severe pain. Only one patient experienced local tumor regression during the chemotherapy, and the incidence of liver metastasis was as low as 13%. CONCLUSIONS: Comparing with the previously reported data in the traditional chemo- and/or radio-therapies, we consider that our method of intra-arterial chemotherapy is quite useful not only for the prolongation of patient's survival but also for improving the quality of life. Thus, this new treatment seems worthy of entering into the prospective randomized study. PMID- 8714227 TI - Regional chemotherapy with hemofiltration: a rationale for a different treatment approach to advanced pancreatic cancer. AB - BACKGROUND/AIMS: Since 1989, thirty-two patients with advanced, intra-abdominal pancreatic cancer were treated with regional chemotherapy in combination with extracorporeal hemofiltration. PATIENTS AND METHODS: Eleven patients had locally advanced, unresectable cancer, and ten had advanced disease with liver metastases. Three patients had developed liver metastases following a radical resection. One patient had an incomplete resection with local residual disease, and a second had developed a local recurrence after a radical resection. One patient had an unresectable cystadenocarcinoma. Five patients had failed prior systemic therapies for unresectable pancreatic cancer. The patients underwent 85 treatments with regional chemotherapy plus hemofiltration, an average of 2.7 treatments per patient. RESULTS: Of 21 patients treated primarily with regional chemotherapy plus hemofiltration, there were two complete responses (9%) and eight partial responses (38%), an overall total response rate of 47%. The average survival for patients with Stage II/III localized, unresectable disease is 13 months and that for Stage IV unresectable disease with liver metastases is 9 months. CONCLUSIONS: Patients with recurrent disease following a radical resection or having failed prior systemic therapies generally had no benefit from regional chemotherapy plus hemofiltration. PMID- 8714228 TI - The role of chemotherapy in the treatment of adenocarcinoma of the pancreas. AB - Adenocarcinoma of the pancreas is a very disappointing disease. By the time of diagnosis, most patients have extensive metastasis, primarily to the liver, peritoneum, and the lymph system. At this point, the disease is usually too advanced to allow resection. Even if resection is performed, the recurrence rate is high. Therefore, there is an urgent need to develop non-surgical therapies. This paper reviews the current role of chemotherapy in pancreatic adenocarcinoma. Various single agent and multiple agent therapies for advanced and locally advanced pancreatic adenocarcinoma are reviewed and discussed. The role of chemotherapy as an adjuvant therapy to surgical resection is also covered in this paper. PMID- 8714229 TI - Small bowel transplantation: current progress and clinical application. AB - Total parenteral nutrition (TPN) is used routinely to maintain patients with the Short Bowel Syndrome (SBS). Until recently, TPN has been the only available therapeutic modality for patients with SBS. Currently, it is the treatment of choice for such individuals and occasionally, when the loss of bowel is extensive, it may be the only way of maintaining life. Unfortunately, TPN is expensive and markedly restrains an individual's lifestyle. Despite the overall success of TPN, the numerous risks associated with its use and the many complications of having an intravenous indwelling for years have served as the stimulus for alternative treatments such as small bowel transplantation (SBT). The first attempts at small bowel transplantation in clinical medicine were by Detterling almost 25 years ago. Patient death or graft loss in these early attempts was caused by the failure to control graft rejection and/or the inability to prevent Graft Versus Host Disease (GVHD). A stimulus for renewed clinical interest in SBT was provided by Starzl et al in 1988 with a report of prolonged graft survival without graft rejection or GVHD in a patient who was the recipient of a multivisceral graft consisting of the entire small bowel and other abdominal organs. Since 1964, 78 Small Bowel transplants have been performed in humans. Several variations of the multivisceral procedure in which the liver and the small bowel constitute the major components of the graft were adopted. The longest survival has been in a child who is still alive with a working graft for more than two years, as reported by Goulet from Paris in 1989. The introduction in SBT of the new immunosuppressive agent FK 506 had provided results which are superior to those achieved with Cyclosporine A (CsA). This latter observation prompted the Pittsburgh group to initiate a large series of isolated and composite intestinal grafts. The remarkable results have demonstrated the clinical utility of intestinal transplantation. This paper will try to summarize the history of the small bowel transplantation until the end of the year 1992, with the current progress in use today. PMID- 8714230 TI - Sphincterotomy of Oddi's muscle through posterior distal duodenum: a modified technique with low morbidity and mortality. AB - BACKGROUND/AIMS: Today, most common bile duct stones are successfully removed with endoscopic and/or laparoscopic techniques; however, in a small number of cases these procedures fail and the stones have to be removed with older techniques. These techniques involve laparotomy and supraduodenal and/or transduodenal common bile duct approaches and are associated with complication rates that may be as high as 12%. We describe in this report a modification of the classic supraduodenal and transduodenal technique that results in substantial decrease of postoperative morbidity and mortality. MATERIALS AND METHODS: Rather than approaching the common bile duct through the anterior wall of the duodenum we reflect the duodenum anteriorly and approach the common bile duct through a 5 10 mm long incision in the posterior wall of the third, horizontal duodenal segment. Prior to the advent of endoscopy and laparoscopy, we used this technique for sphincterotomy and bile duct stone removal in 212 patients. RESULTS: Four to six weeks postoperatively, the common bile ducts were free of stones in 99% of patients, and there were no instances of duodenotomy dehiscence, fistula formation, pancreatitis, infection, hemorrhage, duodenal constriction, ascending cholangitis, or death. CONCLUSIONS: The results shows that sphincterotomy and bile duct stone removal via a duodenotomy in the posterior wall of the horizontal duodenal segment has substantially lower complication rates than if the common bile duct is approached through the anterior duodenal wall. They also suggest that it is the procedure of choice in cases in which endoscopic and/or laparoscopic techniques have failed. PMID- 8714231 TI - Environmental risk factors in inflammatory bowel disease. AB - Besides a genetic predisposition, a causal role of various environmental factors have been taken into consideration in the etiology of inflammatory bowel disease (IBD). The most consistent association of environmental factors so far identified is the association between non smoking and ulcerative colitis (UC) as well a between smoking and Crohn's disease (CD). Other factors such as oral contraceptives, refined sugar, perinatal events, childhood infections, microbial agents, and domestic hygiene have been found to be associated with ulcerative colitis and Crohn's disease but further evaluation is required to confirm the consistency and to define the strength of the association. Recent data also suggest that measles virus may persist in intestinal tissue and early exposure to the virus may be a risk factor for development of CD. The further investigation of environmental factors on IBD and the explanation of their role is expected to open new avenues for basic scientific research and may lead to the development of a more rational approach to the prevention and treatment of IBD. The available data suggest that UC and CD are heterogeneous disorders of multifactorial etiology in which hereditary and environmental factors interact to produce the disease. PMID- 8714232 TI - Prospective study of esophageal motor abnormalities in patients with gastroesophageal disease reflux according to the severity of endoscopic esophagitis. AB - BACKGROUND/AIMS: This prospective study evaluates the peristaltic activity of the thoracic esophagus in a group of patients with gastroesophageal reflux disease grouping them according to the endoscopic findings. MATERIALS AND METHODS: The peristaltic activity and the manometric features of the lower esophageal sphincter were prospectively evaluated in 109 controls and 321 patients with reflux esophagitis, divided according to the degree of endoscopic esophagitis employing Savary's classification. RESULTS: Gastro esophageal sphincter pressure was progressively lower according to the severity of the tissue damage. Motor peristaltic activity of the thoracic esophagus measured by several different parameters was significantly altered in Grade IV esophagitis compared to other groups (p < 0.01). The manometric changes were more pronounced at the distal esophagus compared to the proximal esophagus. The percentage of patients with severe motor disturbances, defined as lower esophageal sphincter (LES) pressure less than 6 mm, less than 50% of peristaltic waves after swallowing and amplitude of contractile waves of less than 50 mmHg were significantly greater in patients with severe esophageal injury (p < 0.001). CONCLUSION: Gastroesophageal esophageal reflux may exhibit a wide spectrum of motor disturbances in the distal part of the esophagus even before the presence of the tissue damage, but it increases in proportion to the severity of endoscopic esophagitis. PMID- 8714233 TI - Gallbladder sensitivity to CCK in duodenal ulcer disease, highly selective and truncal vagotomy. AB - BACKGROUND/AIMS: Following truncal vagotomy, a heightened contractile response of the gallbladder to cholecystokinin (CCK) has been reported in patients. We investigated whether the gallbladder responsiveness to the CCK analog cerulein is also affected in patients with a highly selective vagotomy (HSV) and in duodenal ulcer patients, since most patients had truncal vagotomy for recurrent peptic ulcer disease. MATERIALS AND METHODS: Gallbladder emptying (cholescintigraphy) and plasma cholecystokinin like immunoreactivity (CCK-LI) levels were studied during infusion of graded doses of the CCK analog cerulein. RESULTS: In duodenal ulcer patients (n = 9), patients with HSV (n = 9), patients with truncal vagotomy (n = 9) and control subjects (n = 9), infusion of stepwise increasing doses of cerulein (1-16 ng.kg-1.h-1) induced dose related changes in plasma CCK-LI. In patients with truncal vagotomy, the gallbladder contraction in response to 1, 2 and 4 ng.kg-1.h-1 of cerulein was significantly increased over controls; whereas the gallbladder contraction to cerulein in duodenal ulcer patients and patients with HSV was not significantly different from controls. CONCLUSIONS: Thus, in patients with truncal vagotomy, gallbladder contractile response to CCK is significantly enhanced, possibly due to denervation of hepatic vagal branches since gallbladder contraction after CCK infusion shows no difference between post HSV or duodenal ulcer patients and the controls. PMID- 8714234 TI - Duplication of the gallbladder with advanced adenocarcinoma: diagnostic value of MRI and angiography. AB - We present a very rare case of duplicated gallbladder in which the accessory gallbladder was involved by advanced adenocarcinoma with stones. In this case, MRI was the most useful modality in revealing the cystic formation of the accessory gallbladder with packed stones beside the main gallbladder. Additionally, celiac angiography demonstrated the unusual form of the intrahepatic artery which enveloped the cystic tumor, that might be a diagnostic characteristic of accessory gallbladder. PMID- 8714235 TI - Diagnostic yield of upper gastrointestinal endoscopy in relation to age and gender: a study of 10112 Saudi patients. AB - BACKGROUND/AIMS: The availability of fiberoptic endoscopy has made it relatively easy to evaluate symptoms of upper gastrointestinal disorders with a significant degree of accuracy. MATERIALS AND METHODS: To determine the pattern of upper gastrointestinal lesions in the Saudi population with particular reference to the influence of age and gender, 10,112 patients were studied. RESULTS: No abnormal findings were detected in 26.9%. The frequency of ulcer lesions was 13.7% while non-ulcer, mucosal inflammation was diagnosed with a frequency of 33.7%. In the young (less than 20 years of age), the most common lesion was duodenal ulcer or duodenitis, whereas in the elderly (above 60 years of age), esophagitis, hiatus hernia and esophageal varices were the most frequently diagnosed. The highest mean ages were observed in the group of patients with gastric and esophageal cancers (mean ages of 58.8 and 65.1 years, respectively). There was no gender related differences with regard to the mean age of patients in all the diagnostic categories. CONCLUSIONS: This study provides baseline data on the age distribution of major upper gastrointestinal diseases among the Saudi population. The peculiarities of upper gastrointestinal endoscopic findings in advanced age are clarified. The influence of demographic changes on the pattern of these diseases in Saudi Arabia can be evaluated against our findings in the future. PMID- 8714236 TI - Clinicopathological features of gastric cancer in the remnant stomach. AB - BACKGROUND/AIMS: The clinicopathological features of 34 cases of gastric cancer in the remnant stomach were examined. PATIENTS AND METHODS: Twenty-three patients who underwent primary gastrectomy for benign gastric disease (Group I) and 11 patients who received gastrectomy because of primary gastric cancer (Group II) were studied. RESULTS: The median age at diagnosis of gastric remnant cancer was 62.2 years in Group I and 63.7 years in Group II. The median time interval between the primary operation and the appearance of gastric cancer was 12.6 years in Group I and 6.5 years in Group II. Almost all patients were symptomatic; presenting epigastralgia and constriction. The first diagnostic procedure for these patients was barium examination of the remnant stomach except one patient diagnosed by emergency gastroscopy for hematemesis. Seventy percent of patients in Group I had advanced gastric cancer, most of which were invasive to seromuscular layers, whereas in Group II early gastric cancer was dominant (55%). The prognostic evaluation after the second surgery showed that patients with early stages of cancer had longer survival in both Groups. CONCLUSIONS: These results suggest that early detection of gastric cancer in the remnant stomach by periodical follow up is important and that the application of aggressive surgery for this disease will provide better patient survival. PMID- 8714237 TI - Absence of serologic evidence for hepatitis C virus infection in patients with Hashimoto's thyroiditis. AB - BACKGROUND/AIMS: This paper explores the reported association between Hashimoto's thyroiditis and hepatitis C virus (HCV) infections. MATERIALS AND METHODS: The results of serologic testing for anti-HCV antibody levels were documented in 28 patients (19 female, 9 male) with Hashimoto's thyroiditis and 23 age and gender matched controls with non-Hashimoto's thyroid disorders. RESULTS: The anti-HCV results were negative in both groups. CONCLUSIONS: These findings suggest that despite a reported high prevalence of Hashimoto's thyroiditis in patients with chronic HCV infections, the reverse is not true and hence routine anti-HCV screening cannot be advocated for patients with Hashimoto's thyroiditis. PMID- 8714238 TI - Indications for surgery in the treatment of hepatic hemangioma. AB - BACKGROUND/AIMS: Hepatic hemangiomas are controversial tumors, especially as far as treatment is concerned. This paper analyses a series of 26 cavernous hemangiomas of the liver diagnosed between 1982 and 1993. MATERIALS AND METHODS: A descriptive study is made of cases, their treatment and subsequent follow-up averaging 5 years. RESULTS: A preoperative diagnosis was made in 18 patients. Fifteen of the 26 patients underwent operation, and no deaths occurred. One patient with Kasabach-Merritt syndrome (3.8%) had complications due to rupture of the hemangioma. Postoperative complications were limited to two abscesses (13%). There were no recurrences in the follow-up (average 4.4 years). Three of the 15 surgery patients revealed residual hemangiomas. There were no complications in the 11 non-surgery patients (five of which had giant hemangiomas) during a follow up averaging 5.9 years. One patient initially without pain and with a hemangioma of 7.4 cm became symptomatic, and the hemangioma grew to 9.6 cm; another patient with pain became asymptomatic, with no change in hemangioma size. CONCLUSION: Most hemangiomas can be managed conservatively. Indication for surgery should be assessed in units with experience in liver surgery, where low morbidity and mortality rates can justify the intervention. PMID- 8714240 TI - An audit of ultrasound guided liver biopsies: a need for evidence-based practice. AB - BACKGROUND/AIMS: This report examines complication rates associated with liver biopsies performed under ultrasound guidance. MATERIAL AND METHODS: Following the introduction of a policy of performing all liver biopsies under ultrasound guidance in the radiology department, 168 such procedures were reviewed. RESULTS: The post biopsy bleeding rate 4.2% was high compared to published national figures, which varied widely. CONCLUSION: Our audit did not necessarily support the argument that ultrasound guidance leads to safer liver biopsy, but did support the need for randomized controlled trials of blind and guided liver biopsy to compare safety. The clinical impression gained from the audit was that the change in policy has reduced mortality associated with liver biopsy at this hospital and in the absence of further evidence, the present policy will continue. A further issue raised by the audit was the appropriateness of liver biopsy in terminally ill patients. PMID- 8714239 TI - Hepatic inflow vessels caliber as an index for liver function. AB - BACKGROUND/AIMS: Little is known about the serial change of the hepatic inflow vessels' caliber in the chronically diseased liver. We have analyzed the caliber change in patients with esophageal varices and examined relationship between hepatic inflow vessels diameter and age of patients who had no liver disease. MATERIALS AND METHODS: (i) Diameters of the hepatic inflow vessels was measured on angiogram in five patients with chronic liver disease. They underwent liver angiography twice with an interval of more than 10 months. The caliber change was analyzed. (ii) Thirty-nine patients without chronic liver disease were analyzed to examine vessel diameter change with aging. RESULTS: (i) The left hepatic artery diameter increased in size during the follow-up periods. On the other hand, the portal vein caliber increased in three cases excluding two cases whose Child grade did not change from A. (ii) The hepatic artery and the portal vein diameter had a tendency to become smaller with aging in thirty-nine patients who underwent hepatic angiography and had no liver disease. CONCLUSIONS: Hepatic inflow vessels' caliber change, particularly increase in hepatic artery diameter, has a potential of suggesting the liver function to some extent. PMID- 8714241 TI - Liver transplantation with a twenty-four hour delay and an initial low dose of cyclosporine. AB - BACKGROUND/AIMS: Cyclosporine A based immunosuppression protocols have improved the results of liver transplantation. However, there is no general agreement concerning the most appropriate initial dose of cyclosporine or the precise moment we should start its administration. MATERIALS AND METHODS: Two cyclosporine A administration procedures in liver transplantation were analyzed by means of a prospective study using 91 consecutive patients and dividing them into two groups: Group A: 50 consecutive transplants in which cyclosporine was started since the surgery at 4 mg/kg/day, and Group B: the following 41 consecutive transplants in which cyclosporine was started 24 hours after transplantation at 2 mg/kg/day. RESULTS: Cyclosporine levels were higher in Group A in the first month (without significant differences). There were differences in the need for hemodialysis (14% vs 0%, p < 0.01), in the length of time (h) on mechanical ventilation (147.5 +/- 36 vs 48.7 +/- 15.7, p < 0.05) and in the time (d) spent in intensive care (10.5 +/- 1.6 vs 6.5 +/- 0.8, p < 0.05). There were no differences in the incidence of acute rejection, arterial blood pressure, septic and neurological complications, or in the actuarial survival rate for patients and grafts at 36 months. CONCLUSIONS: Delayed administration of cyclosporine simplifies the treatment of patients in the first 24 hours, it has several beneficial features and does not appear to be associated with a high acute rejection rate. On the basis of these results, our group has adopted delayed and low-dosage cyclosporine procedure. PMID- 8714242 TI - Locoregional immuno-chemotherapy transplenic and transtumoral in vivo immunostimulation. AB - The need for adjuvant therapies for treating the cancer patient has stimulated a great deal of research as well as controversy. Much of the current research is investigating chemotherapy and immunotherapy. One aspect of this research is directed at overcoming drug resistance by the target cells. This paper highlights some of the most recent advances in the fields of chemotherapy and immunotherapy, and discusses the effects of multi-drug resistance protein gene expression. PMID- 8714243 TI - Vater's papilla and periampullary area villous adenoma: personal experience about nine cases and review of the literature. AB - Villous tumor of the duodenum is a rare disease. The authors report their personal experience from 1987 to 1993 with nine cases of duodenal villous tumors. The treatment was pancreatoduodenectomy in four cases, segmental resection of the duodenum in two cases and submucosal excision in three cases. The authors review the literature of the last 15 years in which only 241 cases of duodenal villous tumors are reported; in this review the clinical, diagnostic and therapeutic characteristics of the disease are analyzed. This malignancy presents peculiar characteristics for histologic diagnosis and pathology classification. For this reason, even surgical treatment deserves careful reflection to guarantee a curative procedure and to formulate a positive prognosis. PMID- 8714244 TI - Delayed complications of pancreatic injury: role of CT in detection and management. AB - Pancreatic injury is rare. There is however a recent increase of incidence due to high velocity motor vehicle accidents and urban violence. Diagnosis can be missed during initial assessment. Complications are common due to associated organ injuries. We report on the role of CT in the diagnosis and management of four cases with pancreatic injury of different mechanisms and delayed complications. Advantages and pitfalls of CT are discussed. PMID- 8714245 TI - A new method of duodenum-preserving subtotal resection of the head of the pancreas based on the surgical anatomy. AB - BACKGROUND/AIMS: Duodenum-preserving resection of the head of the pancreas has been performed for benign and, sometimes, malignant diseases of the pancreas. We propose a new procedure of duodenum-preserving subtotal pancreatectomy of the pancreas according to the precise anatomy of the pancreatoduodenal region, especially of the pancreaticoduodenal arteries which provide blood to the duodenum. MATERIAL AND METHODS: After a complete Kocher's maneuver is performed, the pancreas is cut above the portal vein and removed from the third portion of the duodenum, followed by the removal of the posterior surface of the pancreas head from a connective tissue membrane. The main pancreatic duct is identified at its junction with the terminal portion of the bile duct from the posterior surface of the head of the pancreas and is cut at the junction. The pancreas is cut in the line of the ASPD. This line is almost the same as the left side of the common bile duct. The ASPD and the common bile duct should be preserved in this procedure. RESULTS: The reason for leaving part of the pancreas between the duodenum and the anterior superior pancreaticoduodenal artery and the common bile duct is that the artery toward the papilla of Vater runs along the right side of the common bile duct and would be difficult to be preserved with the removal of this part of the pancreas. The most important technique of this procedure is in keeping the connective tissue membrane of the posterior surface of the pancreas intact so as to preserve pancreaticoduodenal arteries and veins, because all the pancreaticoduodenal arteries and veins are situated on this membrane. Complete Kocher's maneuver should cause no problem in this procedure. CONCLUSIONS: Benign lesions as well as low-grade malignancy of the head of the pancreas may possibly be the indication of this procedure. PMID- 8714246 TI - Nonmenstrual toxic shock syndrome: new insights into diagnosis, pathogenesis, and treatment. PMID- 8714247 TI - Antibiotic resistance in Enterococcus species: an update. PMID- 8714248 TI - Resistant nosocomial gram-negative bacillary pathogens: Acinetobacter baumannii, Xanthomonas maltophilia, and Pseudomonas cepacia. PMID- 8714249 TI - Human immunodeficiency virus type 2: human biology of the other AIDS virus. PMID- 8714250 TI - Community-acquired pneumonia. PMID- 8714251 TI - Infections of the eyelid, lacrimal system, conjunctiva, and cornea. PMID- 8714252 TI - Novel plasmid-mediated beta-lactamases in enterobacteriaceae: emerging problems for new beta-lactam antibiotics. PMID- 8714253 TI - Human ehrlichiosis in the United States. PMID- 8714254 TI - Emergence of Vibrio cholerae O139. PMID- 8714255 TI - Hospital-acquired pneumonia. PMID- 8714256 TI - Acute bacterial meningitis in adults. PMID- 8714257 TI - Cysticercosis. PMID- 8714258 TI - Reactivation and exogenous reinfection: their relative roles in the pathogenesis of tuberculosis. PMID- 8714259 TI - Diagnosis and management of invasive aspergillosis. PMID- 8714260 TI - Myeloma: new aspects of biology, prognosis and treatment. PMID- 8714261 TI - Intravascular fasciitis: a case report and review of the literature. AB - A 49 year old man presented with a mass in the inguinal region that for 3 mths had been associated with slight discomfort. Clinically the mass was thought to be an enlarged lymph node. Histology however showed nodular fasciitis. There was also an associated smaller lesion that was histologically identical and which involved the wall of a medium sized vein with protrusion into its lumen. These appearances were typical of intravascular fasciitis. Review of the Royal Brisbane Hospital Pathology files for 12 yrs from 1982, revealed no other such cases. We describe here the clinical and pathological features of this case and review the literature on this rare condition. PMID- 8714262 TI - Proliferating cell nuclear antigen (PCNA) in non-Hodgkin's lymphomas: correlation with working formulation and Kiel classification in formalin-fixed paraffin embedded material. AB - PCNA is a 36-KD proliferating cell nuclear antigen associated with the cell cycle. The immunocytochemical detection of PCNA represents a useful tool for the study of tumor proliferation activity. This study documents the detection of PCNA, using antibody PC 10 in formalin-fixed, paraffin-embedded tissue, and correlates the proliferative activity of the non-Hodgkin's lymphomas (NHL) with histological grading assessed by the International Working Formulation (WF) and Kiel classification. In 92 cases of NHLs we found a strong correlation between the PCNA index and lymphoma grading. Statistically significant differences were also found between the proliferative index (PI) in low and high grade lymphomas according to the Kiel classification (t = 9.519; p < 0.001) and between low, intermediate and high grade lymphomas according to the WF classification (F = 79.01; p < 0.001). In the Kiel classification the mean of low grade lymphomas was 39.5% and of high grade 75.7%. In the WF the average of low grade lymphomas was 29.7%, intermediate 53.1% and high 75.1%. Although the differences among the groups had been significant, we found variations inside each histological subgroup in both classifications. The intermediate lymphomas were the most heterogeneous group, with PI inside the same histologic subtypes coincident with low and high grade lymphomas. Since PCNA may be used as a marker of cell proliferation in clinical studies to estimate the biological aggressiveness of lymphomas, its determination in intermediate grade NHL could be very useful to evaluate individual cases in this group and determine prognosis and probably the appropriate therapy. PMID- 8714263 TI - An immunohistochemical study of p53 protein in cervical intraepithelial neoplasia and squamous cell carcinoma. AB - p53 mutations are known to occur frequently in human cancers where they are considered to be an important event in the stepwise progression towards malignant transformation. It is therefore interesting to compare p53 expression in the uterine cervix for non-neoplastic/metaplastic squamous epithelium, cervical intraepithelial neoplasia (CIN) and invasive squamous cell carcinoma. One hundred and nineteen biopsied and resected specimens of the uterine cervix were stained with an anti-human p53 protein monoclonal antibody by the streptavidin-biotin immunoperoxidase method. Histologically these could be categorized into non neoplastic/dysplastic conditions, including condyloma (34 cases), CIN 1-3 (66 cases) and invasive squamous cell carcinoma (19 cases). Fifty eight per cent (11/19) of the invasive squamous cell carcinomas and 11% (7/66) of the CIN stained positively for p53. Except for 3 cases of invasive squamous cell carcinoma, there was only sporadic intranuclear staining of less than 5% of the cells. No staining was observed in all non-neoplastic/metaplastic squamous epithelial cells. The pattern of p53 staining is significantly different for all 3 categories. However it is undetermined as to whether the positive immunoperoxidase staining is a direct consequence of p53 gene mutation or otherwise. PMID- 8714264 TI - Malignant pulmonary lymphoid disease: case reports illustrating anatomical pattern of disease as a prognostic marker. AB - Although a well documented phenomenon in Hodgkin's disease, malignant pulmonary lymphoid disease in other lymphoproliferative diseases, such as non-Hodgkin's lymphoma (NHL) and chronic lymphocytic leukemia (CLL), is less frequently described. We present 3 patients, 2 with NHL and one with CLL, all demonstrating pulmonary malignant lymphoid involvement. We briefly review the probable mechanism underlying the development of this disease process and identify anatomical distribution of malignant pulmonary lymphoid disease as a prognostic marker in this condition. PMID- 8714265 TI - Angiotropic large cell lymphoma: simultaneous peripheral nerve and skeletal muscle involvement. AB - Angiotropic large cell lymphoma is a rarely encountered disorder characterized by a proliferation of malignant lymphoid cells within vascular lumina. Sites usually affected include the skin and central nervous system, although involvement of multiple other organ symptoms have been described including peripheral nerve and skeletal muscle. Simultaneous involvement of both skeletal muscle and peripheral nerve has rarely been reported. In most of those cases the diagnosis was made only at the time of autopsy. A case of an elderly male with angiotropic large cell lymphoma and in whom the diagnosis was made antemortem on a peripheral nerve and skeletal muscle biopsy is reported. PMID- 8714266 TI - Human papilloma virus and P53 overexpression in carcinomas of the uterine cervix, lower uterine segment and endometrium. AB - Inactivation of the wild type p53 protein through complexing of protein synthesized by specific subtypes of human papilloma virus (HPV) or mutation in the p53 gene in considered to play an important role in cervical carcinogenesis. The association between p53 overexpression and the presence of HPV 16 and 18 DNA was assessed in 29 cervical carcinomas, 15 carcinomas of the lower uterine segment and 30 endometrial carcinomas. In 29 cervical carcinomas (21 adenocarcinomas and 8 adenosquamous carcinomas), 7 cases were positive for HPV 16 DNA while 5 were positive for HPV 18 DNA. Nine cases (31%) showed p53 overexpression. An inverse association was seen between the presence of HPV DNA and the p53 overexpression. In 15 carcinomas of the lower uterine segment, one case was positive for HPV 16 DNA while 2 were positive for HPV 18 DNA. Overexpression of p53 was seen in 8 (53%) carcinomas of the lower uterine segment. Two of 3 HPV DNA positive carcinomas of the lower uterine segment revealed p53 overexpression. HPV DNA was not detected in endometrial carcinomas and p53 overexpression was shown in 12 (40%) cases. The association between HPV DNA and p53 overexpression differs among the tumors arising in the cervix, lower uterine segment and endometrium. PMID- 8714267 TI - The pattern of glomerular disease in New Caledonia: preliminary findings. AB - Two hundred and two renal biopsies from 181 patients in New Caledonia were classified into either primary glomerulonephritis or glomerulopathy associated with systemic disease. These were then compared with 670 similar biopsies from 634 in-patients at Sydney's Royal Prince Alfred Hospital (RPAH). The most prevalent primary glomerular disease among the New Caledonian cases was focal segmental glomerulosclerosis, compared with IgA disease among the RPAH cases. Mesangiocapillary glomerulonephritis, post-infectious glomerulonephritis and minimal lesion nephropathy were all relatively commoner among the New Caledonian biopsies, but the numbers were small. The most prevalent systemic glomerulopathy in the New Caledonian cases were amyloidosis. This was the least common among our RPAH group. Diabetes mellitus and lupus nephritis were also slightly more common in the New Caledonian group. Focal necrotizing/crescentic glomerulonephritis was unusual in the New Caledonian samples, while it was the most common systemic glomerulopathy among the RPAH group. PMID- 8714268 TI - Malignant lymphomas in Papua New Guinea: an immunohistological study of 125 cases. AB - A total of 179 cases of lymphoma, both nodal and extranodal, recorded in the Papua New Guinea Tumor Registry were reviewed and classified by the updated Kiel classification. Fifteen (8.4%) were Hodgkin's disease, 39 (21.8%) probable Burkitt's lymphoma (poorly preserved), 46 (25.7%) Burkitt's lymphoma, 33 (18.4%) Burkitt's-like lymphoma, 36 (20.1%) other B-cell subtypes and 10 (5.6%) T-cell lymphoma. No case of follicular B-cell lymphoma was encountered. PMID- 8714269 TI - Test and teach. Number Eighty. Diagnosis: Malignant eccrine poroma of scalp. PMID- 8714270 TI - Cardiac pathology and adult aboriginal mortality: a coronial study of sudden and external cause deaths in the top end of the Northern Territory in 1990. AB - A retrospective review of coronial records was performed for 179 adult decedents who came to coronial autopsy in 1990 and who had been living in the Top End of the Northern Territory at the time of death. The directly standardized rate of autopsy-examined sudden death was 7.4 times higher for Aboriginal people than for non-Aboriginal people. Coronary atherosclerosis was the cause of 37% (14/38) of the Aboriginal sudden deaths and 52% (16/31) of the non-Aboriginal sudden deaths. Evidence of coronary atherosclerosis rose with age and was seen more frequently in those dying suddenly compared to those dying of external causes. When directly standardized, the rate of autopsy-examined sudden death attributable to ischemic heart disease was 5.5 times higher for Aboriginal people than for non-Aboriginal people. Contrary to a previously published statement, there is considerable overlap between Aboriginal and non-Aboriginal heart weights, with 75% (55/73) of Aboriginal decedents and 84% (89/106) of non-Aboriginal decedents having heart weights between 250 and 500 g. We concluded that the rate of autopsy-examined sudden death from all causes, and specifically from ischemic heart disease, was much higher in Aboriginal people. However in a case of sudden death a presumptive clinical diagnosis of ischemic heart disease prior to autopsy is not justified in either Aboriginal or non-Aboriginal people, due to the high prevalence of other fatal conditions. The diagnosis of pathologically increased heart weight, which is critical in the assessment of sudden death, should be made on the same basis in both Aboriginal and non-Aboriginal people. PMID- 8714271 TI - Antigen retrieval by microwave oven heating for immunohistochemical analysis of bone marrow trephine biopsies. AB - Immunohistochemical analysis of bone marrow trephine (BMT) biopsies with monoclonal antibodies for the analysis of hemopoietic disorders has been hindered by the fixation and decalcification regimens which mask or destroy tissue antigens. This study evaluated the effect of microwave oven treatment on the quality of immunostaining of fixed decalcified trephine biopsies. The aim was to establish whether this method of pre-treatment would enable additional antigens to be detected. Fifty-eight monoclonal and 4 polyclonal antibodies to hemopoietic antigens were assessed to compare no tissue pre-treatment, proteolytic (trypsin) enzyme digestion and microwave oven heating. The microwave heating of the sections was performed by placing them in a boiling solution of 0.01M tri-sodium citrate for a total of 10 mins. Following microwave heating 14 antibodies that previously showed no reactivity in BMT biopsies gave positive staining and 9 antibodies previously known to detect antigens in the absence of pre-treatment gave enhanced staining. Other antibodies showed no staining improvement with microwave heating and some failed to give a positive reaction by any of the pre treatment methods. Antigen retrieval utilizing microwave oven heating can expose antigenic sites for antibody binding in bone marrow trephine sections. However not all antigens are retrieved and there is variation between epitopes on the one molecule and their ability to be exposed by microwave heating. Utilizing antigen retrieval methods, the range of antibodies applicable to BMT sections is greatly expanded enabling the immunophenotypic analysis of the majority of hemopoietic disorders. PMID- 8714272 TI - Donor leukocyte infusions in the treatment of chronic myeloid leukemia in relapse post bone marrow transplantation. AB - A 25-yr-old Caucasian man presented in 1988 with Philadelphia chromosome (Ph) negative, bcr-abl rearranged, chronic phase chronic myeloid leukemia (CML). He was treated with human leukocyte antigen matched sibling allogeneic bone marrow transplantation but relapsed 5 yrs later. At this time he was given donor leukocyte infusions from the original bone marrow donor, seeking an immune anti leukemic effect. This treatment induced graft versus host disease and severe bone marrow aplasia, requiring immunosuppression and repeat donor marrow infusion (without prior conditioning). Graft versus host disease was controlled and full donor hematopoiesis was restored, resulting in complete eradication of the leukemic clone at a molecular level. The patient remains in complete clinical and molecular remission and off all immunosuppression 24 mths later. This emphasizes a potentially powerful graft versus leukemia effect in CML. PMID- 8714273 TI - Rapid and sensitive detection of anti-Ro (SS-A) antibodies by indirect immunofluorescence of 60kDa Ro HEp-2 transfectants. AB - Anti-Ro (SS-A) antibodies are important diagnostic markers for primary Sjogren's syndrome and systemic lupus erythematosus, but their detection by indirect immunofluorescence (IF) in the diagnostic laboratory is hindered by the low cellular abundance of 60kDa Ro protein (Ro60). The approach we used to overcome this problem was to transfect and over-express the Ro60 gene into HEp-2 cells. In this study we have used a mixture of Ro60 transfectants and untransfected HEp-2 cells (HEp-Ro60) as a substrate for IF-antinuclear antibody (ANA) testing in a hospital laboratory. Screening of 240 routine serum specimens identified 14 Ro transfectant-positive sera which were confirmed by counterimmunoelectrophoresis (CIE); 3 of these sera were ANA-negative on untransfected cells and regular HEp 2. A comparison of HEp-Ro60 and regular HEp-2 showed strong concordance of the different ANA patterns between the 2 substrates. No increase in background staining was observed on the Ro transfectants when reacted with normal human sera. A comparison between HEp-Ro60 and CIE for 53 sera from patients with primary Sjogren's syndrome showed that HEp-Ro60 were a sensitive and specific substrate for detection of anti-Ro antibodies. Masking of positive Ro transfectants was observed rarely in sera containing multiple ANA specificities, but the Ro60 staining on these transfectants were unmasked at higher serum dilutions. We conclude that HEp-Ro60 are a suitable substrate for IF-ANA in the routine laboratory and that they have the additional advantage over regular HEp-2 slides of being able to detect anti-Ro in ANA-negative sera. HEp-RO60 are also a valuable confirmatory test for sera giving equivocal precipitin reactions or ELISA results. PMID- 8714274 TI - The occurrence of macrophage-like cholera toxin uptake cells in the intestinal villi of suckling rats. AB - An oral administration of cholera toxin (CT. 10m g) caused diarrhea in infant rats ranging in age from 1 to 14 days. After administration of the toxin a time sequence study was carried out using highly sensitive immunohistochemical procedures. CT was exclusively incorporated into a type of macrophage-like (ML) phagocytic cell. These cells were identified within the intestinal epithelium of rats suffering choleraic diarrhea. After 2 hrs cells taking up the toxin markedly increased in number and were found in both the mucosa and the lamina propria mucosae. After 4 hrs a small number of ML cells containing CT were still present in the mucosal epithelium, but were no longer observed in the lamina propria. Two kinds of monoclonal antibodies against rat macrophages were used to gain a clue as to the cytological characteristics of ML cells. ED1- or ED2-positive macrophages were demonstrable in the lamina propria and submucosa of the small intestines from control rats. In CT-treated rats a considerable number of cells positive for CT and ED1, or CT and ED2 antisera, were found within the epithelial cell layer and the lamina propria of intestinal villi. It is suggested that many ML cells responsive to CT, if not all, are ED1 and ED2 macrophages and are resident in the villous lamina propria where they can migrate to uptake CT in the intestinal lumen. CT B-subunit and heat-labile toxin (LT) B'-subunit from a mutant strain Escherichia coli were given to the rats in order to know the onset mechanism of toxin uptake. It seems likely that the toxin receptor, GM1 ganglioside, participates in the initiation of CT-uptake mechanism. A possible role of the intestinal ML cells was discussed. PMID- 8714275 TI - Evaluation of the Roche Amplicor polymerase chain reaction system for detection of Mycobacterium tuberculosis complex in specimens. AB - Roche Diagnostic Systems Inc. have recently developed a commercial PCR (Amplicor) for direct amplification of Mycobacterium tuberculosis Complex (MtbC) from sputum and bronchial washes. Detection of MtbC specific sequence is achieved by hybridization with an oligonucleotide probe. The aim of this study was to compare the Amplicor PCR system for detection of MtbC with microscopy, culture and an in house PCR method. The commercial assay correctly identified 35/37 microscopy positive specimens compared to 34/37 with the in-house method and 7/13 microscopy negative, culture positive samples compared to 2/13 with the in-house procedure. Negative PCR results were obtained for 32 culture negative specimens and 6 specimens which yielded mycobacteria other than MtbC, indicating a specificity of 100%. The sensitivity of the commercial assay was determined to be approximately 10 organisms compared to 100 organisms with the in-house method. The Amplicor PCR system is specific, sensitive and easy to perform. It also has the advantages of being standardized and quality controlled. PMID- 8714276 TI - Plesiomonas shigelloides and sucrose-positive Edwardsiella tarda bacteremia in a man with obstructive jaundice. PMID- 8714277 TI - Clostridium tertium bacteremia: 2 cases and review. AB - Clostridium tertium bacteremia is unusual, seen most often with gastrointestinal disease and/or neutropenia. Two cases are described. The first was a 19-yr-old female with acute leukemia, who developed gastrointestinal symptoms and C. tertium bacteremia while neutropenic. The second was a 57-yr-old female with quiescent ulcerative colitis, who presented with fever, rigors and epigastric pain. Four organisms including C. tertium were isolated from blood cultures. This patient responded to broad spectrum antimicrobial therapy, whereas the first patient required the addition of specific agents to recover. C. tertium is aerotolerant and thus can be misidentified as a Bacillus or Corynebacterium spp. Our isolates had a distinctive Gram stain morphology, were catalase negative and failed to sporulate aerobically--this aided in the recognition of this significant Gram-positive bacillus. PMID- 8714278 TI - Comparison of PCR/nucleic acid hybridization and EIA for the detection of Chlamydia trachomatis in different populations in a regional centre. AB - Culture on McCoy cell monolayers has been accepted as the reference method for the detection of Chlamydia trachomatis. Recent studies have shown that polymerase chain reaction (PCR)/nucleic acid hybridization based methods have increased sensitivity over culture while still retaining specificity. In situations where organism viability is of concern, due to factors such as transportation delays, culture is inappropriate. Regional laboratories therefore have not been able to utilize the reference method and have been forced to use less reliable methods. The aims of our study were to assess the feasibility of performing PCR to diagnose infections due to C. trachomatis in a regional laboratory using a new commercial kit--Amplicor (Roche Molecular Systems, Branchburg, NJ) and to compare the current enzyme-immunoassay (EIA) based-methods used in our laboratory (VIDAS [bioMerieux Vitek, Hazelwood MO] and IDEIA [Novo Nordisk Diagnostics, Cambridge, UK]) against PCR. Thirteen positive Amplicor specimens were found in 267 urine specimens collected from asymptomatic adolescent males and females. All 13 were confirmed positive using major outer membrane protein gene PCR (MOMP). VIDAS and IDEIA showed 100% correlation to each other but only detected 5/13 positives. Of 140 consecutive patients attending the regional sexual health clinic, 13 were Amplicor positive, 11/13 MOMP positive and 10/13 positive by VIDAS. Five of 254 patients attending the hospital antenatal clinic were positive by Amplicor, all being confirmed by MOMP. No PCR inhibition was detected in a random sample of 100 varied negative Amplicor tests using a modification of the Amplicor kit. No contamination was experienced. The Amplicor kit was shown to be suitable for use in the routine clinical laboratory with minimal disruption to workflow. For regional laboratories this kit should provide more accurate results than EIA based methods, particularly in the detection of asymptomatic persons. PMID- 8714279 TI - Bartonella (Rochalimaea) quintana causing fever and bacteremia in an immunocompromised patient with non-Hodgkin's lymphoma. AB - A 48-yr-old man with stage IV non-Hodgkin's lymphoma, became neutropenic following chemotherapy and developed a fever. His blood cultures were processed to enhance the yield of fastidious bacteria. A slow-growing, capnophilic Gram negative rod was isolated. The febrile episode was treated with cefotaxime, imipenem and vancomycin and resolved. The bacterial isolate was identified as Bartonella (Rochalimaea) quintana by 16S-rDNA gene sequencing. The isolate showed 99.8% sequence homology with the type strain. This is the first isolation of Bartonella (Rochalimaea) quintana from a bacteremic patient in Australia. This bacterium is a fastidious Gram-negative rod requiring prolonged culture for its isolation. Patients with culture-negative pyrexia, especially immunocompromised patients, may need to be investigated for infection with this agent. PMID- 8714280 TI - The frequency of co-infection with Neisseria gonorrhoeae and Chlamydia trachomatis in men and women in eastern Sydney. AB - The frequency of simultaneous infection with Neisseria gonorrhoeae and Chlamydia trachomatis in men and women in Eastern Sydney was examined prospectively over a 21 month period in patients from whom appropriate specimens were obtained. In men gonococcal disease (113 cases) and chlamydial infections (124 cases) occurred in approximately equal frequencies, but there were only 4 instances (3.5%) where C. trachomatis was detected at the same time as N. gonorrhoeae. Demographic data and phenotypic profiles of the gonococci isolated suggested that much of the gonorrhea occurred in homosexual men who are generally considered to have a low incidence of chlamydial disease. Gonococcal disease (34 cases) was far less common than chlamydial infection (112 cases) in women, but co-infection with chlamydia was noted in 6 (17.6%) of the cases of gonorrhea (p < 0.01). The current recommendation for the routine addition of tetracyclines to gonococcal treatment regimens for men requires revision. PMID- 8714281 TI - FNB diagnosis of breast carcinoma associated with HIV infection: a case report and review of HIV associated malignancy. AB - A 48 yr old HIV seropositive female presented with a right breast mass and bilateral axillary lymphadenopathy. Fine needle biopsy (FNB) revealed an adenocarcinoma with abundant mucin production and features suggestive of a cribriform and micropapillary ductal carcinoma in situ (DCIS). Histopathological examination of the tumor confirmed an invasive mixed colloid carcinoma with extensive DCIS. There have been 4 previous reports in the literature of breast carcinoma associated with HIV seropositivity. This case initially diagnosed by FNB is the first case reported in Australia. In spite of the somewhat more favourable histological type of breast carcinoma, this tumor shows numerous unfavourable prognostic factors and has had an aggressive clinical course with relapse of disease in the contralateral breast and distant metastases within 4 wks of surgery, probably related to the patient's immunodeficiency. PMID- 8714282 TI - Squamous cell carcinoma of the distal urethra in a patient with congenital hypospadias. AB - A previously well 43-yr-old man with congenital hypospadias presented with a fungating lesion of 2 mths duration and surrounding the ventral urethral opening. After biopsy showed this to be a squamous cell carcinoma, a definitive procedure of partial penile amputation and urethroplasty was performed. This is believed to be the first reported case of squamous cell carcinoma of the distal urethra in conjunction with hypospadias. PMID- 8714283 TI - Primary intracerebral small lymphocytic non-Hodgkin's lymphoma with plasmacytoid differentiation, associated with prominent extracellular proteinaceous deposits. AB - A case of primary intracerebral non-Hodgkin's lymphoma in a 67-yr-old immunocompetent female is presented. The histopathological diagnosis was supported by immunohistochemical, flow cytometric and electron microscopic findings, and by clinical staging. This tumor is unusual in its morphological features of a low grade, small lymphocytic lymphoma with plasmacytoid differentiation (Working Formulation Classification), and its association with extensive, local, extracellular, proteinaceous deposits. Primary central nervous system non-Hodgkin's lymphomas are briefly discussed and it is postulated that the extracellular proteinaceous deposits in this case originated from immunoglobulins secreted by the neoplastic cells. To our knowledge the massive degree of local immunoglobulin deposition present in this primary central nervous system lymphoma has not been previously reported in the literature. PMID- 8714284 TI - Dementia pugilistica in an alcoholic achondroplastic dwarf. AB - Dementia pugilistica is classically seen in boxers. We describe the case of a 33 yr-old achondroplastic dwarf who developed the pathological hallmarks of the condition, probably as a result of chronic occupational trauma. Dementia pugilistica has not been previously described in achondroplasia. PMID- 8714285 TI - Development of collagenous colitis in a cecoplasty of a patient with interstitial cystitis. AB - We present the case of a 45-yr-old female patient with a 23 yr history of interstitial cystitis that multiple surgical procedures, including a ceco cystoplasty, were unable to cure. After a cystectomy was performed, histological examination of the bladder showed the development of collagenous colitis. Although the etiology of both interstitial cystitis and collagenous colitis is unknown, this report postulates an association between these two conditions. PMID- 8714286 TI - An overview of epithelio-mesenchymal transformation. AB - Epithelium is the tissue phenotype of early embryos and primitive adults of the chordate phylum. A second tissue type, however, is produced by epithelial mesenchymal transformation (EMT) in higher chordates, such as vertebrata. Mesenchymal cells have the ability, which true epithelia do not, to invade and migrate through the extracellular matrix (ECM) to create dramatic cell transpositions. The first-formed or primary mesenchymal cells in amniote vertebrates migrate from the primitive streak to differentiate into the mesodermal and endodermal epithelia. Definitive mesenchyme with connective tissue and muscle potentials arises from the epithelial mesoderm at about the same time as the neural crest mesenchyme forms from the ectoderm. Later on in embryogenesis. EMT is used to remodel unwanted epithelia, such as that of the palate medial edges. We discuss the mechanisms by which epithelial cells transform into mesenchyme and vice versa. On the one hand, cells activate putative mesenchymal master genes, turn off epithelial genes, and acquire motility machinery that allows them to interact in 3 dimensions (3D) with ECM via actin cortex while sliding their endoplasm into their new front ends. On the other hand, primary mesenchymal cells can reactivate epithelial regulatory genes, such as E-cadherin, turn off the motility machinery for invading ECM, and reexpress apical-basal polarity. We review the genes, such as FSP1, src, ras, and fos, that are activated in cells transforming to mesenchyme and the genes their neighbors activate to induce EMT, such as those for TGF beta, NT-3, and sonic hedgehog. Suspension in 3D collagen gels can induce adult epithelium to undergo EMT; alpha 5 beta 1 integrin is activated on surfaces in contact with collagen, including apical surfaces that do not normally express integrins. In vivo, it is possible that pathological manipulations of a cell's environment likewise induce EMT. Of the examples we give, the creation of invasive metastatic carcinoma cells by EMT is the most fearful. Interestingly, transfection of either metastatic cells or normal embryonic fibroblasts with the E-cadherin gene converts them to the epithelial phenotype. It may be possible in the future to manipulate the tissue phenotype of diseased cells to the advantage of the animal. PMID- 8714287 TI - Cell-cell adhesion molecules in epithelial-mesenchymal transformations. AB - The functional units in most inductive and morphogenetic processes in the embryo are not single cells, but rather collectives of interacting cells that give rise to the tissues and organs. Cell adhesion molecules (CAMs) are involved in defining cell collectives and their borders as they interact during inductive events in morphogenesis. The expression patterns of CAMs are highly dynamic and changes are known to occur during epithelial-mesenchymal transformations. Alterations in CAM expression are correlated with changes in morphology. Conversely, experimentally induced changes in morphology result in changes in CAM expression. The structure, function, distribution, and control of CAM gene expression are presented in this review, and discussed with regard to their importance to normal developmental processes, particularly epithelial-mesenchymal transformations. PMID- 8714288 TI - Recent insights into ligand binding, activation and signalling by integrin adhesion receptors. AB - In recent years, analyses of the structure and function of membrane-intercalated adhesion molecules have shown them to play key roles in determining cellular phenotype. As expected, adhesion has an important role in regulating cellular positioning, but there is also compelling evidence that information transduced via adhesion molecules affects the differentiation status of cells. Cell surface adhesion molecules can be classified into a number of gene families, including immunoglobulins, cadherins, selectins, proteoglycans, and integrins. All of these types of molecule are co-expressed on most cells, and therefore the overall contribution of adhesion to cell phenotype is likely to be a net effect of the individual contributions of each of these groups. In this review, we will focus on the role of the integrins, which appear to be particularly important mediators of cell migration and adhesion-dependent intracellular signalling. A great deal is now known about the extracellular faces of integrins, including their structure and ligand-binding mechanisms, and in recent years, our knowledge of integrin-dependent signalling via cytoplasmic domains has improved considerably. An emerging picture is one of a dynamic family of receptors than can be expressed in different states of activation. Alterations in activity are apparently mediated by conformational changes that can be induced from both outside and inside cells. In turn, these changes in activity have concomitant consequences for adhesion and signalling. PMID- 8714289 TI - Molecular interactions in the submembrane plaque of cell-cell and cell-matrix adhesions. AB - Adhesion of cells to their neighbors or to the extracellular matrix has multiple effects on cell shape, dynamics and fate. The most obvious and direct one is the assembly of single cells into ordered multicellular tissues and organs. This process requires specific transmembrane adhesion molecules which mediate the binding to the external surface, cytoskeletal filaments which attach to the cytoplasmic faces of the adhesion site, and a submembrane plaque which interconnects the two. The co-assembly of these junctional domains is essential for the formation of stable cell adhesions with the proper mechanical properties. In addition, adhesive interactions have prominent, global consequences on cell behavior and fate, affecting such processes as differentiation, growth and survival. To gain insight into the molecular basis for both the local and global effects of adhesive interactions, we have chosen to focus on one specific junctional domain, the submembrane plaque of microfilament-bound adhesions, namely cell-cell and cell-matrix adherens junctions. Based on both biochemical and morphological evidence we would like to propose that the junctional plaque plays a key role in mediating and regulating transmembrane junctional interactions and adhesion-dependent signaling. It offers multiple modes of linkage between the cytoskeleton and the membrane, and its assembly can be controlled at either the biosynthetic or posttranslational levels. Furthermore, recent data demonstrate that the submembrane plaque is involved in the transduction of transmembrane signals. We will show that this structure is the residence of an array of signaling enzymes (mostly kinases), that its structure and composition may be affected by activation of various signaling systems, and that adhesion itself may activate specific signal transduction pathways. PMID- 8714290 TI - Epithelium-mesenchyme transition during neural crest development. AB - The neural crest is the organ system whose presence defines vertebrates. The onset of migration of neural crest cells is an archetypal epithelium to mesenchyme transition (EMT), and this event identifies the cell lineage. Little is known yet of the establishment of the neural crest, although the zinc finger gene Slug seems to be involved in specifying EMT competence. The details, especially the temporal order of events in neural crest EMT, vary between different species and between different axial levels, but several important features have emerged from observations in situ and experiments in vitro and in vivo. EMT seems to be strongly associated with decrease in cell-cell adhesion, and particularly with loss of N-cadherin on the surface of neural crest cells at the time of onset of migration. The related adhesion molecule T-cadherin is also present, but correlated changes have not yet been described, while the unrelated adhesion molecule N-CAM also declines on neural crest cells, but with a time course unrelated to EMT. The extracellular matrix is also important: EMT-related changes in matrix receptor (i.e. integrin) activity are recorded in avian crest cells, while the nature of the matrix itself changes in urodele amphibians. Changes in cell shape and in cell motility also occur at the time of EMT, consistent with changes in the cytoskeleton. These concerted changes can be triggered by TGF-beta family growth factors, of which dorsalin-I appears particularly important. These may act through pathways involving controlled alterations in phosphorylation to effect the complex of responses that make up EMT. Although much remains to be understood, the spatiotemporal definability of this system makes it a very useful model for studying EMTs in general. PMID- 8714291 TI - Epithelio-mesenchymal transformation during formation of the mesoderm in the mammalian embryo. AB - The earliest example of epithelio-mesenchymal transformation during embryonic development is the generation of the third germ layer, the mesoderm, from the epiblast (or primitive ectoderm), which marks the beginning of gastrulation. Although it has been regarded as most likely that the principles of this transformation in invertebrates and lower vertebrates also apply to amniotes, morphological and molecular details of mesoderm formation in birds and, in particular, in mammals, which may support this assumption, have only recently been clarified. This chapter thus brings together the light- and electron microscopical morphology of epithelio-mesenchymal transformation during initial mesoderm formation in the mammalian embryo. Cellular differentiation during this process with regard to the cytoskeleton, cell adhesion molecules and the extracellular matrix are also covered as are cell kinetic studies and the candidate growth factors and genes most likely to be involved in the regulation of mesoderm formation in mammals. Finally, a model is presented which summarizes these morphological and molecular changes and which links the promoting and inhibiting influences of regulatory factors to some of the changes observed during epithelio-mesenchymal transformation. PMID- 8714292 TI - [In memoriam Prof. Lev Germanovich Leibson]. PMID- 8714293 TI - [The identification of repetitive sequences and internal symmetry in the primary structure of N-acetylglucosaminyl-1-phosphotransferases]. AB - UDP-GlcNAc: Dol-P GlcNAc-1-P-transferases (GPT) are the high conservative family of enzymes which catalyze the first reaction of Dol-PP-GlcNAc2Man9Glc3 biosynthesis. This oligosaccharide is necessary for protein N-glycosylation taking place in the endoplasmic reticulum of eucaryotic cells. The analysis of amino acid codon roots made it possible to identify symmetrical segments in the primary structure of GPT. The centres of symmetry of these segments are mainly localized in potential membrane-spanning hydrophobic regions of proteins participating in dolychol-binding regions forming both catalytic and allosteric domains of GPT. Two types of repeating homologous amino acid sequences of yeast GPT are revealed by using graphic method of primary structure analysis. The first type is mainly characterised by the prevalence of hydrophobic amino acids and forms potential transmembrane domains. Hydrophilic amino acids forming hydrophilic loop in C-terminal region of GPT are present in the second type. These sequences consist of 15-18 amino acids residues and occupy more than two thirds of enzymic molecule. PMID- 8714294 TI - [The species characteristics of the intramolecular structure of hemoglobin in vertebrates]. AB - Using fluorescence spectroscopy species peculiarity of hydrophobic and microviscosity degree in central and peripheral molecular areas of the main electrophoretic haemoglobin fractions in representatives of different vertebrate classes gas been studied. It is established that central areas of haemoglobin molecule are characterised by twice lesser variability of size and hydrophobic degree and 2.4 times greater variability of package strength as compared with peripheral areas of protein globule. PMID- 8714295 TI - [The effect of excitatory amino acid antagonists on postsynaptic potentials in the motoneurons of the frog Rana ridibunda]. AB - The effects of excitatory amino acids on postsynaptic potentials (PSP) generated by activation of three synaptic inputs in isolated frog Rana ridibunda spinal cord motoneurons were investigated. PSP evoked by dorsal root (DR) and reticular formation (RF) stimulation were blocked by kynurenate (0.5-1 mM) and CNQX (1 mkM). However, the resistivity of RF PSP and DR PSP to kynurenate and CNQX was recorded in 7 and 4 cases respectively. APV blocked the late phase of DR and RF PSP, but did not decrease the amplitude of the early phase. Lateral column (LC) PSP generated in the same cell by LC stimulation was not decreased by kynurenate or CNQX at all. LC PSP was blocked by APV in 1 of 6 cells and it was blocked by the mixture of strychnine and APV in 2 of 6 cells. DR and RF PSP resistant to kynurenate were also blocked by the mixture of strychnine, picrotoxin and APV in 3 of 17 cells. Strychnine itself did not block LC PSP or resistant DR and RF PSP. Moreover, LC PSP was depressed, whereas RF and DR PSP were potentiated while glutamate was added into superfusing solution. PMID- 8714296 TI - [A comparative histochemical study of cytochrome oxidase activity in the somatosensory and auditory brain centers in the normal rat and after exposure to superhigh-frequency electromagnetic fields]. AB - Using histochemical method mitochondrial cytochrome oxydase (CO) in acoustic and somatosensory centers of rat brain has been studied to reveal CO activity distribution in norm and after impulse-modulated high-ultra-high frequency influence. After ultra-high frequency influence the increase of enzymic activity in a number of regions of rat brain centers with relationship to processing ecologically important sensory signals is revealed. PMID- 8714297 TI - [The vasopressin- and oxytocinergic systems of rat strains selected for their capacity to acquire active avoidance]. AB - Vasopressin- and oxytocinergic structures of the hypothalamus were stained immunochemically. Volume of the nucleoli of the neurosecretory cells in the paraventricular, supraoptic and anterior commissural nuclei was calculated as a measure of neurohormone synthesis. The amount of the neurosecretory material in the neurohaemal organs (external median eminence and posterior pituitary) was estimated cytophotometrically. It was shown that both synthesis and secretion of the neurohormones were higher in intact KLA rats as compared to KHA rats. In three days after inescapable electroshock opposite changes in synthesis and secretion of both oxytocin and vasopressin were revealed: a rise in KHA rats and a fall down in KLA ones. It is suggested that differences between KHA and KLA rats in the reaction of vasopressin- and oxytocinergic systems to stress is due to different strategy of rats behavior at electroshock. PMID- 8714298 TI - [Nontraditional "large-cell" neuroendocrine formations (accessory nuclei) in the brain of Anamnia and Amniota]. AB - Using immunochemical PAP-method nonapeptidergic neuroendocrine formations in the hypothalamus and adjacent brain areas of fishes (the sterlet Acipenser ruthenus, the shark Scylliorhinus canicula), amphibians (the frog Rana temporaria), reptiles (the snake Natrix natrix), mammals (rats and dogs) and human have been studied. In Amniota and human accessory nuclei (AN) in addition to main "magnocellular" nuclei (supraoptic, postoptic and paraventricular) were discovered. Two AN, circular and dorsolateral ones, were found in snakes, and circular, dorsolateral, forniceal and extrahypothalamic AN were revealed in rat, dog and human brain. In Anamnia, sharks and frogs, in contrast to sterlets, the dorsolateral sub-nucleus inside preoptic nucleus was identified. AN similarity in the phylogenetic row of vertebrates and mechanisms of AN creation in phylo- and ontogenesis were discussed. PMID- 8714299 TI - [The effect of water deprivation on the mother-fetus system at different stages of pregnancy in rats]. AB - Female Wistar rats were exposed to stress (water deprivation) during 3 days starting from the 8th, 14th or 19th day of pregnancy. In all cases serious changes in the mother's organism were observed and they were manifested in the essential delay of body mass increment. The main indexes of reproductive function (the number of alive new-born rats, relationship between males and females in broods) did not change. The signs of lagging behind in development were revealed for experimental groups of new-born rats (less body mass, delay of bone formation) and they were the most pronounced in the group exposed to stress at the 14-17th day of pregnancy. In postnatal period no essential differences between experimental groups and control in terms of teeth appearance, hearing onset, as well as in the indexes of physical endurance were observed. Behavioral reactions of progeny in the "open field" appeared to be changed only for the generation of female rats exposed to stress in the second period of pregnancy and were different on the exposures as of the 14-17th and 19th-22nd day of pregnancy. The results obtained have been discussed in the context of views about critical periods of development. PMID- 8714300 TI - [The reproductive function of young male silver foxes Vulpes vulpes after long term selection for domesticated behavior]. AB - Sperm morphology of spermaries, testosterone plasma level and sexual activity of young silver fox males selected for domestic behavior have been studied. It is established that a number of spermatozoa in semen of males from selected population is lower as compared with the control one, whereas abnormal spermatogenesis level is significantly increased. The testosterone plasma level in males after a female introduction increased to the same extent in both groups. Sexual activity in males from selected population was lower than in the control one during the first mating season. The data obtained suggest a depressing effect of selection for domestic behavior on spermatogenesis and sexual activity in the young silver fox males and point to the hereditary changes of some chains of reproductive functions of males under the selection for domestic behavior. PMID- 8714301 TI - [Cognitive activity during the recognition of a different number of objects in the adolescent chimpanzee]. AB - The ability of adolescent chimpanzee (5-7 years old) to recognise and generalize flat pictures of discrete amounts has been studied. A set of tasks (19 signal pairs) with pictures of different number of objects was used. Situational conditional reflexes of chimpanzee were developed in conditions of free behavior. Chimpanzee were taught to choose a large number of pictures. Reversion ability at conversion of signal value of irritants was examined. It is shown that chimpanzee are able to transfer the early developed knowledge to choose larger picture to larger number of objects. It points to rather developed function of generalization, and ability to make abstracts. Resolution of tasks with reversion appeared to be difficult for chimpanzee. Correlation between the ability to solve tasks of different complexity, latent period of choice and etiological indexes is revealed. Both common regularities and individual differences in formation of complex behaviour of chimpanzee are established. PMID- 8714302 TI - [The neurobiological bases of acoustical communication. A comparative physiological review of structures and mechanisms]. PMID- 8714303 TI - [The submerged electro-olfactogram of the clawed toad Xenopus laevis]. AB - Relative effectiveness of 17 amino acids solutions (5 x 10(-4) mol/l) for olfactory receptors of the clawed frog has been determined by measuring underwater (EOG) amplitude in ventro-medial and lateral areas of the nose. It was revealed that hydrophilic amino acids (cysteine, glycine, threinine, tyrosine) produced larger EOG in ventro-medial area as compared with hydrophobic ones (alanine, valine, leucine, phenylalanine, proline, tryptophan). Hydrophobic amino acids were more effective in lateral area. It was shown that olfactory receptors in ventro-medial area but not in the lateral one in regard to stimulating amino acids were similar to olfactory receptors of the frog Rana temporaria. Amino acids which induced the greatest increase of mobility in the frog Xenopus laevis (the reactions to these amino acids were similar to the effects of food stimuli) produced the largest EOG in ventro-medial area. PMID- 8714304 TI - The impact of the quality of family planning services on contraceptive use in Peru. AB - Through linkage of a Demographic and Health Survey to a situation analysis, this article explores whether current contraceptive use in Peru is affected by the service environment in which a woman resides. The investigation focuses explicitly on the impact of the quality of family planning services and finds that, net of personal and household characteristics, a significant, albeit small, effect exists for one specification of quality in the total sample and for the other specification a nearly significant (p = .053) effect exists. The analysis reveals that contraceptive prevalence would be 16 to 23 percent greater if all women lived in a cluster with the highest quality of care compared with the lowest. Methodological problems that arise in measuring quality of care at the cluster level and in linking quality to individual contraceptive use are also addressed. PMID- 8714305 TI - The effect of operations research on program changes in Bangladesh. AB - This article is based on the ten-year experience of an operations research project in Bangladesh. It assesses how, and under what circumstances, research based advice and results of pilot projects contribute to change in large-scale public programs. It discusses project research on issues facing the national family planning program: recruitment and training of field-workers; delivery of injectable contraceptives; management information; field-workers' use of service registers; field supervision; satellite clinics; and contraceptive user fees. These issues are used to illustrate the advantages and disadvantages of a long term institutionalized project, and to describe the diversity of means for communication with policymakers. The analysis shows that research, policy decision, and implementation can occur in any sequence. Policy advice that disrupts long-standing power relationships and organizational culture takes a great deal of effort to implement. Operations research can produce useful changes in organizational behavior, even when large-scale problems remain. PMID- 8714306 TI - Reweighting DHS data to serve multiple perspectives. AB - Information about health and family planning infrastructures is collected through the service availability module (SAM), an important feature of the Demographic and Health Surveys (DHS) conducted in many developing countries. The DHS samples were designed to provide a representative sample of households and women of reproductive age. Using the weights routinely provided with DHS data sets, service accessibility can be described straightforwardly at the individual and household levels. However, without further adjustment, SAM data do not provide a representative picture of service delivery at the community, or primary sampling unit, level, where the data are collected. This report proposes a methodology for reweighting the SAM data, using rural data from the Egypt DHS as an illustration, so that available family planning facilities at this level may be usefully characterized at little additional cost. PMID- 8714307 TI - The impact of outreach on the continuity of contraceptive use in rural Bangladesh. AB - In 1978, the Bangladesh family planning program launched a national program of outreach services that continues to the present. Young married women were hired and trained to visit women in their homes, offer contraceptive services, provide information, and support sustained use over time. This report uses data from two rural districts to assess the effect of the household visitation program on the continuity of contraceptive use. Results of a multivariate analysis show that household outreach has had a pronounced net effect on the continuity of contraceptive use throughout the study period and that the magnitude of this effect has increased with time. This finding suggests that sustained contraceptive use continues to benefit from home-based outreach even after a decade of service encounters. Policy implications of this finding are discussed. PMID- 8714308 TI - Ideal-family-size and sex-composition preferences among wives and husbands in Nepal. AB - This study tests the hypothesis that, in Nepal, measures of ideal family size mask an underlying preference for sons, making some people willing to have families larger than their ideal. Existing evidence suggests that men are likely to have stronger preferences for sons than are women. This research uses empirical evidence to examine the hypothesis that husbands are more willing than their wives to pursue the birth of sons at the cost of an increasingly large completed family size. A Multiple-response Fertility Preference Scale was developed to test these propositions among a sample of couples. The methodology was successful in demonstrating differential patterns of decisionmaking between husbands and wives that are otherwise obscured by more simplistic, single response measures (for example, ideal family size). The results indicate that husbands are consistently more willing than their wives to pursue the birth of sons at the expense of larger family sizes, and that the birth of daughters is not pursued to a similar degree by wives or husbands. PMID- 8714310 TI - The multiple personality debate. PMID- 8714309 TI - Indonesia 1994: results from the Demographic and Health Survey. PMID- 8714311 TI - The multiple personality debate. PMID- 8714312 TI - The multiple personality debate. PMID- 8714313 TI - Catecholamines in attention-deficit hyperactivity disorder: current perspectives. AB - OBJECTIVE: To provide an update on the "catecholamine hypothesis" of attention deficit hyperactivity disorder (ADHD). METHOD: Recent work examining the measurement of the norepinephrine, epinephrine, and dopamine systems in ADHD and normal subjects is reviewed and discussed in the context of recent neuroimaging and animal studies. RESULTS: While data clearly indicate a role for all three of the above neurotransmitters in ADHD, a hypothesis suggesting "too much" or "too little" of a single neurotransmitter will no longer suffice. The central norepinephrine system may be dysregulated in ADHD, such that this system does not efficiently "prime" the cortical posterior attention system to external stimuli. Effective mental processing of information involves an anterior "executive" attention system which may depend on dopaminergic input. The peripheral epinephrine system may be a critical factor in the response of individuals with ADHD to stimulant medication. CONCLUSION: A multistage hypothesis is presented which emphasizes the interaction of norepinephrine, epinephrine, and dopamine in modulation of attention and impulse control. PMID- 8714314 TI - Humpty Dumpty. PMID- 8714315 TI - Prediction of adolescent affective disorder: effects of prior parental affective disorders and child psychopathology. AB - OBJECTIVE: To examine the role of major parental and child diagnostic factors in predicting episodes of serious affective disorder in adolescents in a nonreferred sample. METHOD: The sample included 139 youngsters (average age 14 years at enrollment) drawn from a health maintenance organization and evaluated at two points in time 4 years apart. Both parents and adolescents were assessed using structured diagnostic instruments scored according to criterion systems. Parent and child lifetime diagnoses identified in the first assessment were used to predict the onset of episodes of serious affective disorder in the adolescents which occurred between the first and second assessment. RESULTS: Stepwise multiple regression analyses of the significant univariate factors showed that the most powerful predictors of episodes of affective disorder were total number of diagnoses the adolescents received prior to first assessment, lifetime duration of parental major depressive disorder, and total number of lifetime nonaffective disorders of the parents. CONCLUSION: Broad risk factors from different domains best predict episodes of affective disorder in children and adolescents. PMID- 8714316 TI - Depressed mothers: they don't always look as bad as they feel. AB - OBJECTIVE: The relationship of maternal depression to self-report and observational data was investigated in a sample of depressed (n = 30) and nondepressed women (n = 32) and their 3-year-old children. Depression characteristics (diagnostic subtype/remission status) were related to maternal self-report and mother-child interactions. METHOD: Mothers completed standardized questionnaires and the dyad was observed in the laboratory. Observations were rated for maternal behavior and child attachment. RESULTS: There were no differences between depressed and nondepressed groups on observational measures; depressed mothers' self-report was consistently more negative. Within the depressed group, women with more severe/chronic depression showed behavioral differences but were no different from less depressed women on self-report measures. Women in remission improved for self-report, but not for observational data. CONCLUSIONS: The consequences of maternal depression vary depending on type of depression, severity, chronicity, current mood status, and on how the impact of depression is measured. PMID- 8714317 TI - Thyroid hormone concentrations in depressed and nondepressed adolescents: group differences and behavioral relations. AB - OBJECTIVE: To examine thyroid hormone concentrations and the influence of these hormones on mood and problem behaviors in adolescents with depression. METHOD: The sample included 21 depressed adolescents and 20 matched control adolescents. Blood was drawn to measure thyroid-stimulating hormone (TSH), free thyroxine (FT4), thyroxine (T4), and triiodothyronine (T3). Major depression (MD), attention deficit (AD), and obsessive-compulsive (OC) symptom scores were abstracted from the Diagnostic Interview Schedule for Children. Total behavior problem scores from the Youth Self-Report also were obtained. RESULTS: Paired analysis revealed there were no significant group or gender differences or group by gender interactions for TSH, T4, or T3. For FT4, however, there were significant group differences (p = .008) showing lower concentrations in depressed adolescents than control subjects, suggesting that the former might be functionally hypothyroid. Although there were no significant correlations of TSH with any of the psychological measures obtained, in the depressed group correlations were negative (although not always significant) with FT4 and total behavior problems (r = -.40, p = .09), as well as with symptom scores of MD (r = .25, p = .288), OC (r = -.56, p = .011), and AD behaviors (r = -.57, p = .008). Higher numbers of symptom scores of OC and AD were related to lower concentrations of FT4. CONCLUSIONS: FT4 concentrations were lower in depressed adolescents. These findings suggest a relationship between negative behaviors and dysfunction of the hypothalamic-pituitary-thyroid axis in adolescents with depression. PMID- 8714318 TI - Structural abnormalities in brain magnetic resonance images of depressed children. AB - OBJECTIVE: Brain magnetic resonance images (MRIs) of 65 children and adolescents who were hospitalized with depressive disorders (DD) were compared with the brain MRIs of 18 hospitalized psychiatric controls (PC) without a depressive disorder. METHOD: Volumetric analyses were used to measure frontal lobe volumes (FLV), lateral ventricular volumes (VV), and total cerebral volumes (CV) for all subjects. To correct for differences in absolute cerebral volume associated with different body and head size, the ratios of FLV/CV and VV/CV were used to compare differences between the two groups. A multivariate analysis was used to control for the effects of several independent variables (age, sex, diagnosis). RESULTS: Significant differences were seen in the FLV/CV ratio and the VV/CV ratio when the results were compared between the two groups (DD versus PC). The DD group had a significantly smaller FLV/CV ratio (t = 2.148, df = 79, p = .035) and a significantly larger VV/CV ratio (t = -2.093, df = 79, p = .040). CONCLUSION: The findings are consistent with previous reports in depressed adults and may implicate the frontal lobes in the pathogenesis of early-onset depressive disorders. PMID- 8714319 TI - Case study: electroconvulsive therapy in adolescents. AB - OBJECTIVE: Recognizing the limited experience with electroconvulsive therapy (ECT) in adolescent psychiatric patients, the authors evaluated the efficacy, indications, and benefit of the treatment in this population. METHOD: The records of patients in the ECT rosters at University Hospital at Stony Brook between 1983 and 1993 were reviewed. Thirteen adolescents between the ages of 16 and 18 years who received ECT were identified. ECT followed the institution's standard adult protocol. RESULTS: Ten patients were evaluated as responders, and three as nonresponders. Patients with diagnoses of an affective illness, unspecified psychosis, and catatonia benefited the most. Some improvements were seen in the patients with schizophrenia. Eight patients (62%) were contacted by telephone for follow-up. Five remain asymptomatic 3 years after the treatment, and three relapsed within 12 months despite maintenance pharmacotherapy. CONCLUSION: Within the limitation of record review and partial follow-up, ECT was found to be an effective and useful procedure in the treatment of adolescents with severe psychiatric disorders. PMID- 8714320 TI - Comparison of diagnostic criteria for attention-deficit hyperactivity disorder in a county-wide sample. AB - OBJECTIVE: To examine teacher-reported prevalence rates for attention-deficit hyperactivity disorder (ADHD) based on DSM-III-R and DSM-IV in the same population. METHOD: Teachers completed questionnaires in which they rated all their students on all DSM-III-R and DSM-IV symptoms for disruptive behavior disorders except for seven conduct disorder symptoms but including seven symptoms screening for anxiety or depression. This constituted all children in kindergarten through fifth grade in a middle Tennessee county during the 1993 1994 academic year (16 schools, 398 teachers, and 8,258 children). Also included were questions about the children's diagnosis of ADHD, treatment with stimulants, and the presence of behavior or academic problems. RESULTS: The prevalence rates were 7.3% for ADHD (DSM-III-R); 11.4% for ADHD, total (TOT); 5.4% for ADHD, inattentive type (AD); 2.4% for ADHD, hyperactive-impulsive type (HI); and 3.6% for ADHD, combined type (CT). Factor analysis identified five factors: opposition/defiance-conduct, inattention, hyperactivity/impulsivity, anxiety/depression, and stealing-truancy. The rates of problems differed mostly between ADHD-AD and ADHD-HI (40% versus 80%) for behavior and (75% versus 23%) for academics. Few (15% to 40%) had an ADHD diagnosis or stimulant treatment (21% to 32%). CONCLUSION: DSM-IV criteria are likely to increase the prevalence of this disorder in comparison with DSM-III-R rates, but they may better characterize its heterogeneity. PMID- 8714321 TI - Validity of DSM-IV ADHD predominantly inattentive and combined types: relationship to previous DSM diagnoses/subtype differences. AB - Since 1980, three different diagnostic nomenclatures have been published regarding attention-deficit hyperactivity disorder (ADHD). These changing conceptualizations and diagnostic criteria have generated considerable confusion. OBJECTIVE: To examine the multidimensional DSM-IV ADHD criteria in relation to how children and adolescents with a previous DSM-III ADD diagnosis or a DSM-III-R ADHD diagnosis are diagnosed according to DSM-IV criteria. METHOD: Children whose original diagnoses were according to DSM-III and DSM-III-R criteria received retrospective diagnoses according to DSM-IV criteria. RESULTS: Predominantly inattentive (n = 30) and combined types (n = 26) were compared on their previous DSM-III and DSM-III-R diagnoses and on demographic, behavioral, cognitive, and comorbidity variables. Predominantly inattentive and combined type diagnoses corresponded with DSM-III ADD/WO and ADD/H diagnoses, respectively. The DSM-III-R ADHD diagnosis did not correspond with either DSM-IV subtype. Children with the combined type diagnosis had more externalizing codiagnoses, and their parents reported more externalizing, delinquent, and aggressive behaviors. Children with the predominantly inattentive type had more math learning disability codiagnoses. CONCLUSION: Results support a multidimensional conceptualization of ADHD. There exists close correspondence between the DSM-III ADD/WO type and the DSM-IV predominantly inattentive type and between the DSM-III ADD/H type and the DSM-IV combined type. PMID- 8714322 TI - Objective measurement of hyperactivity and attentional problems in ADHD. AB - OBJECTIVE: To precisely describe movement abnormalities in seated children with attention-deficit hyperactivity disorder (ADHD) while they were engaged in a continuous performance task (CPT). METHOD: Diagnoses were made by using structured interviews (Schedule for Affective Disorders and Schizophrenia for School-Age Children-Epidemiologic Version) and DSM-IV criteria. Movement patterns of 18 boys with ADHD (9.3 +/- 2.4 years) and 11 normal controls (8.6 +/- 1.8 years) were recorded using an infrared motion analysis system that tracked the position of four markers 50 times per second to a resolution of 0.04 mm. RESULTS: Boys with ADHD moved their head 2.3 times more often than normal children (p < .002), moved 3.4 times as far (p < .01), covered a 3.8-fold greater area (p < .001), and had a more linear and less complex movement pattern (p < .00004). They responded more slowly and with greater variability on the CPT. Complexity of head movement and variability in response latency significantly correlated with teacher ratings. A predefined composite of movement and attention discriminated 16 of 18 patients from 11 of 11 controls. CONCLUSIONS: The relative inability of boys with ADHD to sit still can be objectively verified, and "fidgeting" appears to consist of more frequent, larger amplitude, whole body movements. PMID- 8714323 TI - Predictors of persistence and remission of ADHD into adolescence: results from a four-year prospective follow-up study. AB - OBJECTIVE: To evaluate the predictors of persistence and the timing of remission of attention-deficit hyperactivity disorder (ADHD). METHOD: Subjects were 6- to 17-year old Caucasian, non-Hispanic boys with and without ADHD. DSM-III-R structured diagnostic interviews and blind raters were used to examine psychiatric diagnoses, cognitive achievement, social, school, and family functioning at a 4-year follow-up assessment. RESULTS: At the 4-year follow-up assessment, 85% of children with ADHD continued to have the disorder and 15% remitted. Of those who remitted, half did so in childhood and the other half in adolescence. Predictors of persistence were familiality of ADHD, psychosocial adversity, and comorbidity with conduct, mood, and anxiety disorders. CONCLUSIONS: The findings prospectively confirm that the majority of children with ADHD will continue to express the disorder 4 years later. For a minority of children, ADHD was a transient disorder that remits early in development. In addition, we have shown that persistence of ADHD is predictable. Familiality, adversity, and psychiatric comorbidity may be clinically useful predictors of which children with ADHD are at risk for a persistent disorder. PMID- 8714324 TI - Carbamazepine use in children and adolescents with features of attention-deficit hyperactivity disorder: a meta-analysis. AB - OBJECTIVE: In the United States approximately 750,000 children receive psychostimulants to treat attention-deficit hyperactivity disorder (ADHD); up to 25% may not respond. The purpose of this study was to evaluate the reports in the international literature concerning the efficacy of carbamazepine (CBZ) in children with ADHD features by means of meta-analysis. METHOD: A review of the world literature located 29 reports that dealt with behavior problems, activity levels, and CBZ in children. Of these, only 10 reports provided sufficient or pertinent information for the meta-analysis. RESULTS: In all seven open studies, therapeutic responses were significant (ranging from p = .05 to .0001, two-tailed t test). Meta-analysis using weighted variables revealed a significant positive correlation (r = .88; p < .02) between duration of treatment and positive outcome. In three double-blind placebo-controlled studies, treatment effects for CBZ's superiority over placebo ranged from p = .07 to .0001. A meta-analysis of these three studies revealed that CBZ was significantly (p = .018) more effective than placebo at controlling target symptoms. CONCLUSIONS: Despite the general lack of attention that CBZ has received for treating ADHD, there is preliminary evidence that CBZ may be an effective alternate treatment in children with features of ADHD. PMID- 8714325 TI - Twenty-one-month follow-up study of school-age children exposed to Hurricane Andrew. AB - OBJECTIVE: To explore the 21-month course of posttraumatic stress symptomatology (PTSS) and psychological morbidity in 30 school-age children (7 to 13 years) after exposure to Hurricane Andrew. METHOD: Pynoos' Posttraumatic Stress Disorder Reaction Index and Achenbach's Teacher's Report Form were administered at 8 and 21 months after Hurricane Andrew. RESULTS: At 21 months 70% of the children endorsed moderate-severe PTSS. The reduction in PTSS was greater for boys than girls. Psychopathology as measured by the Teacher's Report Form increased over the 19-month period. Boys demonstrated significant increases in internalizing symptoms and in Withdrawn, Anxious/Depressed, Social Problems, and Attention Problems scales, and girls showed a significant increase in the Anxious/Depressed scale. CONCLUSIONS: Twenty-one months after exposure to Hurricane Andrew, there were continuing high levels of PTSS and evidence of increasing emotional and behavioral problems. While girls sustained higher levels of PTSS, boys demonstrated higher indices of other psychopathology. The enduring effects of disaster associated with secondary stressors and "traumatic reminders" continue to be etiologically important for continuing psychological morbidity. PMID- 8714326 TI - Seven-year follow-up of child survivors of a bus-train collision. AB - OBJECTIVE: To assess the long-term effects of a traumatic bus-train collision and to examine the effect of levels of exposure and immediate reactions on long-term adjustment. METHOD: Seven years after the accident, 389 subjects, all doing compulsory army service, filled out self-report questionnaires assessing symptoms of posttraumatic stress disorder (PTSD), psychiatric symptomatology, and military functioning. In addition, subjects were questioned about their immediate reactions to the traumatic event. RESULTS: The most highly exposed subjects reported the highest levels of somatization, depression, phobic anxiety, and psychoticism and more PTSD symptoms. Acute stress symptoms and manifestations of fear immediately after the accident were strongly related to long-term maladjustment CONCLUSIONS: The results suggest that the investigation and assessment of long-term adjustment after traumatic events should take into account both contextual factors, such as the level of exposure to the event, and personal factors, such as the victims' immediate reactions. PMID- 8714327 TI - Relocation after a disaster: posttraumatic stress disorder in Armenia after the earthquake. AB - OBJECTIVE: To explore the relationship between exposure to the earthquake in Armenia on December 7, 1988, and relocation from the disaster zone, and the subsequent development of posttraumatic stress disorder (PTSD), depression, and behavioral difficulties in children. METHOD: The PTSD module of the Diagnostic Interview for Children and Adolescents-Revised and the Depression Self-Rating Scale were administered to 25 children, aged 11 through 13 years, who had high exposure to the earthquake and remained in the earthquake city. They were compared with a demographically similar group of 24 children exposed to the earthquake who were relocated to another city after the earthquake and 25 nonexposed children. For each child the mothers responded to the Child Behavior Checklist and the teachers responded to the Teacher's Report Form. RESULTS: The hypothesis that relocated children would present with less PTSD, depression, and behavioral problems was not confirmed. Both groups of children with high exposure to the earthquake, one remaining in the earthquake city and one relocating, demonstrated significantly higher rates of PTSD, depression, and behavioral difficulties than the comparison group. There were no differences between the relocated children and those who remained in the earthquake zone. CONCLUSION: Children who were relocated after a natural disaster did no worse than children who remained in the disaster zone. Relocation should be considered as an alternative after catastrophic natural disasters in situations where resources are so limited that rebuilding cannot take place for an indefinite period of time. PMID- 8714328 TI - The Khmer Adolescent Project: III. A study of trauma from Thailand's Site II refugee camp. AB - OBJECTIVE: To determine the prevalence rates of posttraumatic stress disorder (PTSD) and depression in a sample of 99 Cambodian youths, aged 18 to 25 years, living in the Site II refugee camps along the Thai-Cambodian border; to compare these rates to data collected in a similarly aged sample of Cambodian refugees living in the United States; and to illustrate the findings with case vignettes and a brief description of the refugee camp at Site II. METHOD: The senior author describes the main features of life in the Site II camp while being employed in one of its medical clinics. A Khmer translated version of the depression section of the Schedule for Affective Disorders and Schizophrenia for School-Age Children Epidemiologic Version and the PTSD section of Diagnostic Interview Schedule for Children and Adolescents were used in interview format by trained bilingual research assistants. Khmer versions of the Beck Depression Inventory and the Impact of Events Scale were also administered. RESULTS: The enduring nature of PTSD was evident in this sample of Khmer youths who had survived the Pol Pot regime as children. Similar rates of Pol Pot-related PTSD were found when compared to rates from the US sample. Subclinical forms of PTSD were found in those who reported their worst trauma during life in the camp, while the full PTSD syndrome was associated with those who reported trauma occurring during the earlier Pol Pot regime. Extremely high rates of depressive disorder were found which were interpreted as related to the repatriation back to Cambodia as this study was undertaken. CONCLUSION: PTSD in this sample appears to be specifically related to earlier war trauma, while depressive symptoms appear more related to recent stressors. As with other findings from the Khmer adolescent project, this study reaffirms the strong connection between the diagnosis of current PTSD and earlier war trauma in an additional sample of youths at Site II, Thailand. Depressive symptoms, on the other hand, appear to be related to the vicissitudes of recent stressful events in this refugee population. PMID- 8714329 TI - Lessons from a hematology/oncology psychosocial conference. PMID- 8714330 TI - The interplay of national, state, and local policy in financing care for drug affected women and children in California. AB - Recent prevalence studies in California indicate that perinatal alcohol and other drug use remains a serious issue for large numbers of women and their children. In response, national, state and local policymakers have taken steps to address the problem, including increasing funding for treatment services. To gauge the impact of policy attention to this problem, the Center for the Vulnerable Child at Children's Hospital, Oakland, California, surveyed state and local administrators of programs that serve drug-affected women and children in California. Information collected included the scope of program services, indicators of access, and sources of program funding. Surveyed programs were funded through federal, state, county, and foundation sources. Despite new policy and funding initiatives to serve this population, the study found wide gaps between the kinds of services that are believed to be appropriate for drug affected women and children and the service system that currently exists. Problems in access to care included long waiting times, exclusion of women from programs based on their pregnancy or parenting status, and exclusion of drug exposed children from programs with medically based eligibility criteria. Program funding sources appeared to impede access, as traditional federal, state, and county funding sources do not support programs that are comprehensive, family centered, and easily accessible to these women and children. Analysis of the study data suggests that policymakers consider new approaches to promote access to care for these underserved women and children, particularly now as Congress and the states redesign health and social service funding mechanisms and delivery systems. PMID- 8714331 TI - Access to substance abuse treatment for homeless women of reproductive age. AB - Homeless women in the United States comprise a subpopulation at high risk for substance abuse, with rates of substance use disorder ranging from 16% to 67%. Despite the need for treatment that such high rates imply, relatively few substance-abusing homeless women avail themselves of formal treatment. The fact that they tend not to utilize formal treatment services is especially problematic among homeless women of reproductive age, who are not only themselves at risk of health-related problems but who place their fetuses and children in danger of multiple negative consequences. The imbalance between treatment need and treatment access suggests that homeless, substance-abusing women are facing severe barriers to care. Although identifying barriers to their treatment access is crucial if this imbalance is to be remedied, very little empirical research has been done in this area. This article presents an overview of current knowledge about barriers to substance abuse treatment for women generally and for homeless women specifically, and proposes a comprehensive empirical strategy for redressing the lack of information on homeless women's access to substance abuse treatment. PMID- 8714332 TI - Options for recovery: promoting success among women mandated to treatment. AB - In recent years imprisonment has been used increasingly for a wide range of nonviolent and petty offenses committed by women. Among incarcerated women, particularly those who are pregnant or parenting, substance use and its deleterious consequences are often exacerbated by imprisonment. Women who have been identified as chemically dependent are also at high risk for losing custody of their children. In California, the Options for Recovery (OFR) treatment program provided an alternative to incarceration or relinquishment of custody of children for chemically dependent pregnant and parenting women. This three-year pilot project offered alcohol and other drug abuse treatment and case management to these women, and included special training and recruitment of foster parents for their children. Findings from a three-year, multimethod evaluation study showed that women who were mandated to OFR treatment programs were more likely to successfully complete treatment than women who had enrolled in OFR voluntarily. An economic analysis of the costs associated with women in OFR compared with the combined costs of incarceration and alcohol and other drug abuse treatment produced a ratio in favor of OFR. Additionally, some innovative service alternatives for women mandated to treatment were developed during the project. The impact of such changes have implications for improving women's and family health. PMID- 8714333 TI - Challenges in moving from a traditional therapeutic community to a women and children's TC model. AB - With the advent of specialized programs for addicted women and their children, some of the traditional methods used by therapeutic communities have been undergoing significant changes. This article examines the issues that are important for treatment providers to consider as they move from individual client and community orientation to a mother-child/family-centered and community approach. The major adaptations suggested are divided into three categories: structural design issues, including living arrangements and models of childcare; treatment issues, including acting-out behavior by the children; and staff and training issues, including staff composition. PMID- 8714334 TI - Involuntary smoking cessation: a treatment option in chemical dependency programs for women and children. AB - Recent research has indicated that a relationship exists between nicotine addiction and the abuse of other substances. This relationship, as well as the severe impact of nicotine addiction on the health of women, their children, and the developing fetus, provides the basis of a rationale for developing chemical dependency programs for women where smoking is not allowed. Involuntary smoking cessation (ISC) programs have been tried recently in mixed-gender and male-only programs, and have met with strong resistance from clients. In most published reports this resistance was strong enough to force the programs to eliminate the ISC policy. This article describes the development of an ISC program at a residential substance abuse treatment center for pregnant and postpartum women and their children. It traces the evolution of tactics to defuse resistance and enlist client support for the program. The development of techniques to measure the effectiveness of the program are also presented. Preliminary results indicate that a properly designed program can be instituted at a residential treatment center for women without excessive program disruption and with positive results. PMID- 8714335 TI - Family functioning of perinatal substance abusers in treatment. AB - This prospective study examined family characteristics of perinatal substance abusers enrolled in an intensive outpatient treatment program both during and after treatment. Data is provided on family functioning in three areas: current family, family of origin, and love/partner relationship, and at six time points up to 24 months postdischarge. Standard family assessment measures, including the Self-Report Family Inventory (SFI) and Family-of-Origin Scale (FOS) were utilized to measure current family functioning and family of origin health, respectively. A new instrument, the Relationship Assessment Form (RAF), was used to measure the level of unhealthy, codependent behaviors in the subject's love/partner relationship. A mixed models repeated measures ANOVA was used to determine if subjects' family functioning changed over time. Multivariate and univariate methods examined differences between the subjects in the current study and previously reported means for the SFI and FOS. Results were that perinatal substance abusers' perception of the overall health of their families of origin remained stable over time. Subjects rated their families of origin as more pathological than community adults, but similarly to adults presenting for psychotherapy. Although current family functioning was relatively stable, SFI scores that changed worsened over time after treatment. Perinatal substance abusers scored more pathologically at intake than normal, nonclinic families, but were similar to families presenting for family therapy on the SFI. In contrast, enduring changes of greater magnitude occurred in the subjects' self-reported codependent behavior over time. All RAF scores that changed moved in the direction of healthier, more autonomous functioning. Change and stability in aspects of family functioning may be due to specific factors of the treatment program, which are discussed. Standard family measures may need further refinement for use in this population. Recommendations for clinical programs and future research are provided. PMID- 8714336 TI - Making evaluation work in a substance abuse treatment program for women with children: Olivia's House. AB - Olivia's House represents the development and implementation of an innovative substance abuse treatment program for drug-dependent, primarily African-American women with children. This article discusses the prevalence of problems related to women and drug abuse; describes the philosophy and continuum of care of Olivia's House; delineates the program, process, outcome, dissemination, and training components of the evaluation protocol; presents preliminary evaluation findings as they relate to the overall goals of Olivia's House: and points out the obstacles to the evaluation of a residential drug abuse treatment program from women and children. This article is targeted to substance abuse service providers and evaluators challenged with the effective delivery and accountability of meaningful treatment services to persons with substance use disorders. PMID- 8714337 TI - Women offenders and reentry issues. AB - Women parallel men in their profile of physical disease, psychosocial configuration, addictive patterns, and criminal deviance. For women offenders in particular, the prison environment reinforces a victim role that originated in childhood and adolescence. In addition, such settings discourage both emotional expression (except for aggression) and responsibility, since basic needs such as food, lodging, and clothing are provided. Incarcerated women have significant treatment issues, including the lack of social and vocational role definition, psychological dependence and psychic craving, poorly developed social skills, and conflicts in social, family, and intimate relationships. This article describes the unique psychoeducational and skills-training needs of women offenders as they adjust to community living, and outlines specific treatment interventions that have proven to effect successful patient outcomes. Case studies are used to illustrate the reentry experiences of three women offenders with distinct backgrounds. One example illustrates how a woman who had been involved in the criminal justice system for 24 years overcame her addiction and self-confidence issues. A second case study profiles an offender with three children who had experienced sexual trauma during her childhood and adult years. A third case reports on an African-American woman whose crack-cocaine addiction resulted in the birth of a drug-exposed son. The treatment model tested in all three cases emphasized the practical and often overlooked treatment issues of incarcerated women. PMID- 8714338 TI - Sexual abuse, physical abuse, and posttraumatic stress disorder among women participating in outpatient drug abuse treatment. AB - Findings from a prospective, longitudinal study of 182 women and 148 men in outpatient drug abuse treatment programs indicate that women are significantly more likely than men to experience sexual and physical abuse. Sexual and physical abuse are associated with higher levels of posttraumatic stress disorder (PTSD) symptomatology. Moreover, women are more likely than men to possess an array of psychological factors that predict relapse to drug use after treatment, including low self-esteem, depression, anxiety, and suicidal behavior, among others. But contrary to expectation, PTSD is not associated with relapse to drug use, nor are women more likely than men to relapse within a six-month posttreatment interval. Further analysis indicates that while women have more psychological risk factors associated with relapse, they are more likely than men to engage in the treatment process. Engagement in treatment, notably frequent participation in group counseling, appears to mitigate the higher risk of relapse for women. PMID- 8714339 TI - Normal sleep architecture in infants and children. AB - Because the collection of normative sleep data in the pediatric age group largely depends on technical issues, this article discusses various aspects of sleep recordings in infants and children. Polygraphic monitoring of sleep and waking behavior contributes to a better description of maturational stages in children, from the prenatal period to the end of adolescence. This article reviews the general characteristics of normal children's sleep and the environmental factors that influence these maturational processes. The use of polygraphic recordings to identify neurological deficits and to predict behavioral and neurological outcome in infants and children, however, does not hold great promise. The importance of polysomnographic evaluation is admitted for some major clinical conditions, such as risks for abnormal breathing or narcoleptic attacks. PMID- 8714340 TI - Obstructive sleep apnea in infants and young children. AB - The obstructive sleep apnea syndrome (OSAS), is a common cause of morbidity during childhood. Childhood OSAS usually stems from adenotonsillar hypertrophy. OSAS in infants is usually related to craniofacial anomalies. Other risk factors include obesity and neuromuscular disease. Symptoms include snoring and difficult breathing during sleep. Infants may have problems with feedings and experience failure to thrive. Definitive diagnosis is made by polysomnography. Normative polysomnographic parameters vary with age; thus age-appropriate norms must be used. In contrast to adults, children often manifest a pattern of persistent partial airway obstruction during sleep, rather than cyclical, discrete obstructive apneas. Most children are cured by tonsillectomy and adenoidectomy. However, some children require further therapy, such as continuous positive airway pressure. PMID- 8714341 TI - The laboratory assessment of daytime sleepiness in childhood. AB - Excessive daytime sleepiness is a disabling symptom of diverse etiologies. This article reviews the various, currently available diagnostic techniques that can be applied in the evaluation of childhood hypersomnolence, along with their merits and limitations. Normal polysomnographic data of children differ from those of adults. Serial studies may be required in evolving childhood narcolepsy for establishing a definitive diagnosis. PMID- 8714342 TI - Clinical neurophysiology of cortical sensorimotor function: yesterday, today, and tomorrow. PMID- 8714343 TI - Automatic Oxford-Medilog 9200 sleep staging scoring: comparison with visual analysis. AB - We report the results of a comparison study between automatic sleep staging using the ambulatory Oxford-Medilog 9200 system and visual analysis in a group of 30 subjects. Sleep analysis was performed during the second nighttime period of each 48-h polygraphic monitoring. The automatic analysis determined a significant underestimation of total sleep time and stage 2 non-rapid eye movement (NREM) sleep and an overestimation of stage 1. No significant differences were found between the two analyses regarding slow-wave sleep and REM sleep. The differences between automatic and visual analysis of stage 2 were correlated positively with the amplitude and duration of K-complexes and negatively with the amplitude and frequency of spindles. Differences in electroencephalographic characteristics of these sleep figures were correlated negatively with age. Our results showed that (a) the Oxford-Medilog 9200 sleep staging system did not improve the scoring of stages 1 and 2 NREM sleep and (b) the degree of differences between visual and automatic analysis are related to the amplitude and frequency of sleep figures, i.e., spindles and K-complexes, used in the scoring of stage 2. PMID- 8714344 TI - The effects of early and late preterm birth on brainstem and middle-latency auditory evoked responses in children with normal neurodevelopment. AB - In preterm and term infants, brainstem and middle latency auditory evoked responses (ABR and MLR) were obtained at 40 and 52 weeks conceptional age (CA) and at 5 years of age. A neurological and neuropsychological evaluation was performed at 5 years of age. To study the effect of preterm birth on the maturation of the ABR and MLR, the preterm infants were divided into early and late preterm groups. Only children with a normal neurodevelopmental outcome at 5 years of age were entered into the study. For ABR, the late preterm group showed significantly longer mean latencies IIc, III, V, and Vc when compared with the term group at 52 weeks CA. There was a trend to longer ABR latencies I in the early preterm group compared with the term group. At 52 weeks CA, the late preterm group showed longer mean interpeak latencies III-I and V-I when compared with the term as well as the early preterm group. At 5 years, the late preterm group showed significantly longer mean ABR latencies IIc and III when compared to the early preterm group. For MLR, the early preterm group showed significantly longer mean latencies of MLR component PO when compared with the term group at 40 weeks CA. At 52 weeks, the late preterm group also had longer mean MLR latencies P0 than the term group. At 5 years of age, the term group showed higher mean peak to-peak amplitudes Na-P0 than the early as well as the late preterm group. To a large extent, the ABR results support the hypothesis that middle ear effusions in combination with retarded myelination of the central auditory pathway are responsible for the ABR differences found between term and preterm infants with a normal neurodevelopmental outcome at 5 years of age. The longer latencies and interpeak latencies found in late preterm infants when compared with early preterm infants might be explained by an augmented vulnerability of the auditory pathway between 30 and 34 weeks CA. The MLR differences found between term and preterm infants might be explained by a difference in the maturation of primary and nonprimary MLR components. PMID- 8714345 TI - Magnetic coil suppression of extrafoveal visual perception using disappearance targets. AB - We used magnetic brain stimulation with a butterfly coil over the occipital lobes to study extrafoveal visual field effects in six subjects. The visual test pattern was a grid of asterisks around a central fixation point, and the target was the disappearance of one asterisk for a single frame of the video monitor. Using single magnetic pulses at stimulator outputs of 55-95%, we noted robust interference effects at latencies < or = 100 ms, peaking at approximately 50-90 ms. Suppression of visual perception occurred with both transverse and sagittal alignments of the coil. When the coil was moved laterally over either occipital lobe, perception of target disappearance was consistently suppressed in the contralateral visual field. Movement of the coil rostrally produced consistent suppression in the lower and middle field, but preferential suppression of the upper field could not be obtained. This altitudinal asymmetry may be correlated with the anatomy of the occipital lobe in relation to the scalp surface. PMID- 8714346 TI - Magnetic coil suppression of visual perception at an extracalcarine site. AB - Perception of extrafoveal visual targets can be suppressed by magnetic stimulation over the occipital lobes, but the site of interference for this and similar phenomena has not been well defined. We modified a previously used technique to determine the locus of magnetic activation. Using butterfly stimulus coils of different sizes and electric field profiles, we determined a scalp position of minimum threshold and a level of stimulator output that produced 50% error rates for each coil. Intersection of the corresponding electric field profiles in air and in a saline model head was similar and identified superficial occipital cortex rather than the primary visual area as the site of perceptual suppression. Less direct analyses involving the distribution of induced electric fields produced the same conclusion. These results suggest specific hypotheses about the effects of magnetic stimulation on visual physiology. PMID- 8714347 TI - Changes in motor unit estimates with aging. AB - Elderly persons usually exhibit some degree of muscle atrophy, together with a reduction in voluntary strength, but there is still argument concerning the nature of the cellular events involved. This issue was reexamined by estimating the numbers and relative sizes of motor units in three limb muscles, using a fully automated system (Galea et al., 1993). In 79 healthy volunteers aged 20-98 years, estimations of motor unit numbers were performed on the thenar, biceps brachii, and extensor digitorum brevis muscles. Motor unit populations were noted to decrease significantly with age in the distal muscles but appeared to remain constant in the biceps. The excitable muscle fiber mass, as reflected in the peak to-peak amplitude and area of the maximum M-wave, was diminished in all three muscles. Although the area of the average motor unit action potential was not significantly different between groups, the ratio of this potential to the M-wave area increased with increasing age. The results suggest that muscle deterioration in the elderly is due to a combination of changes in the muscle fibers and in their nerve supply and that the extent may differ between proximal and distal muscles. PMID- 8714348 TI - [The formation and development of the denaturation theory of injuries and irritation]. AB - The main idea of the denaturation theory (Nasonov, Aleksandrov, 1940) is that the reversible denaturation changes in the most unstable cellular proteins lay in the basis of the similar nonspecific injuries in the cell caused by different agents. Changes of this type also may be developed in the cell as the result of physiological stimuli. The latter suggestion is confirmed by current data about increasing of the conformational flexibility of the protein receptors under specific challenge. Conformational changes of the proteins to "the molten globule state" which is intermediate between native and denaturated states take place in many biological processes. Decrease of the conformational flexibility of the proteins connected with increase of their thermostability also plays an important role in the cell activity at normal and stress conditions. Denaturation theory of the injuries and irritation led to the understanding of the correlation in the ectothermic organisms between life temperature and the level of the conformational flexibility of their proteins which should be supported in the semi-stable state. PMID- 8714349 TI - [The bioenergetic, cytochemical, and staining properties of normal hepatocytes and under functional loads, adaptations to environmental changes and in nonspecific injuries]. AB - Two groups of cells were found among mouse liver cells and among hepatocytes in culture after their fixation and staining: weakly stained cells ("light" cells LC), and strongly stained cells ("dark" cells- DC), the latter making about 30% of population. DC and LC do not differ in their capacity for DNA, RNA and protein synthesis, in contents of proteins, carbohydrates, lipids etc., and also in their ultrastructure. DC differ from LC in lower acidity (pH 6.5) of live cell cytoplasm, and in higher level of mitochondrial fluorescence after staining with Rhodamin 123. The live culture hepatocytes were seen in two alternative states: DC are able to become LC, and vice versa. The amount of DC increases after a high functional cell activation (partial hepatectomy, phlebotomy and hormonal stimulation), and after cell adaptation to modification of cultural medium conditions (modification of pH, hypo- and hypertonia, hyperthermia). In these cases hepatocytes maintain the capacity to granule formation after vital staining with Neutral red. After nonspecific moderate alteration (6% ethanol) of cultured hepatocytes, the acidity of cytoplasm and a temporary elevation of mitochondrial fluorescence may occur, in addition to the increase in the total number of DC. It is suggested that such a cell activation, both in normal physiological conditions and in the case of cell adaptation to modified cultural media, represents a general stress reaction of the cell preceding the "preparanecrosis" state. PMID- 8714350 TI - Optical monitoring of activity in the nervous system: a brief history. PMID- 8714351 TI - [The role of ionic transporters in the long-term regulation of the water content in animal cells. The mathematical model and real lymphoid cells]. AB - Ion and water balance as well as flux balance in the animal cell, equilibrated to anisosmotic solutions, are computed for a cell model supplied by Na,K-ATPase pump, electroconductive ion channels and symporters Na-Cl, K-Cl and NaK2Cl. It is shown how the potency of each of the principal transporters to regulate cell volume depends on such conditions as the state of other ion transporters and channels, initial ion distribution and membrane potential. The obtained data are applied to studying changes in cation and water balance in the Jurkat lymphoid cells equilibrated to hyposmotic solutions. It is concluded that a steady state volume regulation in Jurkat cells, maintained in hyposmotic media 180 mosM for up to 24 h, is achieved due mainly to non-ionic mechanisms. Under the lower osmolarity of the media, 155 mosM, the ionic cell volume regulation occurs. The increase in PCl and/or the decrease in Na-Cl symport (or its equivalent, e.g. Na/H and Cl/HCO3 exchange) are considered as the most probable cause of the long term volume decrease in lymphoid cells. PMID- 8714352 TI - [The Na,K-ATPase pump and blast transformation of human peripheral blood lymphocytes]. AB - A transition of phytohemagglutinin (PHA)-activated human lymphocytes from resting state to proliferation is accompanied by an early and a long-term, delayed increase in ouabain-sensitive Rb influx. The long-term (between 16 and 48 h of PHA action) activation of the Na,K-ATPase pump is concomitant with the enlargement of lymphocytes and precedes the onset of DNA synthesis. When inhibiting the protein synthesis in activated lymphocytes, cycloheximide fails to suppress the early rise in Rb influx; however, it abolishes completely the increase in ouabain-sensitive Rb fluxes after 2 h of PHA stimulation. The inhibitors of transcription, actinomycin D and alpha-amantin, in doses suppressing the increase in the PHA-induced RNA synthesis, do not abolish the elevated Rb influx within 20-24 h of PHA action, and inhibit a growth-related increase in Rb influx during the 2nd day of activation. It is concluded that in mitogen-stimulated human lymphocytes the protein synthesis dependent enhancement of Na,K-ATPase pump activity is different in its nature at different phases of G0/G1/S progression. Unlike the early pump stimulation (0.5-2 h), the late elevation of K influxes associated with the growth of lymphocytes, is due to the regulation of the Na,K-ATPase pump at translational (2-20 h) and transcriptional (after 20 h) levels. PMID- 8714353 TI - [A morphofunctional analysis of the changes in the Golgi apparatus in the epitheliocytes of the frog bladder under conditions of the vasopressin stimulation of water transport]. AB - Structural and chemical peculiarities of the Golgi apparatus elements in granular cells of the normal frog urinary epithelium and under vasopressin stimulation of water transport have been studied with different electron microscopic methods: standard chemical fixation, prolonged osmification, freeze-substitution, freeze fracture, immunocytochemistry, and electron-probe X-ray microanalysis. The structure of the main Golgi elements and its derivatives in normal cells and under the stimulation of water transport has been described. The association of microtubules with the Golgi cisternae was shown. Microtubules are supposed to participate in the support of integrity of the Golgi complex (in normal cells). Under stimulated water transport, depolymerization of microtubules seems to occur, resulting eventually in the Golgi fragmentation. Participation of some specific granules, that are the Golgi derivatives, in the increase of apical membrane water permeability has been shown as the insertion of water channels. Besides, under big water flows, the Golgi cis-cisternae were shown to participate in the formation of large vacuoles containing low potassium. A supposition is put forward that these vacuoles may perform an osmoregulative function in the cell, similar to that of contractile vacuoles of Protozoa. PMID- 8714354 TI - [The characteristics of the transport into the nucleus of conjugates with normal and mutant nucleophilic sequences]. AB - A study was made of the transport of conjugates of a fluorescently labeled protein and a synthetic peptide, corresponding to the nuclear localization sequence of the SV 40 large T-antigen, into the nuclei of cultured human (HeLa and A431) and murine (HER14) cells. A possibility for such conjugates to be transported into the nuclei of digitonin-permeabilized cells, without addition of exogenous cytosol, was demonstrated. A quantitative comparison of the transport levels of constructions with normal or altered (K128T) peptide sequence was performed, and a low selectivity of nuclear transport with this alteration in the digitonin-permeabilized cell system was revealed, whereas constructions with the mutant sequence, when injected into live cells, remained in the cytoplasm. ATP dependent transport of constructions with the mutant sequence into permeabilized cell nuclei was demonstrated, with a considerable part of this transport being NEM-insensitive. A suggestion is put forward that there are several variants of the nucleophilic sequence containing protein transport in the digitonin permeabilized cell system. PMID- 8714355 TI - [The tyrosine kinase activity of the internalized epidermal growth factor receptor in A-431 cells]. AB - The interaction between epidermal growth factor (EGF) and specific membrane receptors in A-431 cells results in stimulation of its tyrosine kinase (TK) activity. The intrinsic tyrosine kinase phosphorylates both the receptor itself and different intracellular substrates. Upon EGF binding, clustering and internalization of EGF-receptor (EGF-R) complexes are observed. In the present study by incubating A-431 cells with EGF at 4 or at 37 degrees C, eluting surface bound EGF we were able to compare the TK-activity of internalized EGF receptors to those localized on the plasma membrane. By the method of the phosphorylation of exogenic substrate poly (Glu/Tyr) 4:1 in vitro it was shown that the total TK activity was equal to 3 microM PO4- incorporated/mg protein/min. Under conditions, when 40% EGF-R complexes were internalized their activity consisted of 46% of the total after inactivation 60% surface-bound receptors. We draw a conclusion that the TK-activity of EGF-receptors is remained during their internalization. PMID- 8714356 TI - [The functional state of denatured proteins: the principles of modelling and the first results]. AB - The aqueous environment near cell membranes should be characterized by a local low pH value (due to attraction of protons by the membrane electrostatic potential) and by a local dielectric constant value (due to proximity of the water-organic boundary). It was proposed to simulate the behaviour of proteins near membranes by their behaviour in water-alcohol mixtures at moderately low pH values. Cytochrome c and retinol-binding protein are shown to be denatured at the values of pH and dielectric constant typical for the aqueous environment near the membrane surfaces. The conformational state of both these proteins under mentioned conditions is similar to the molten globule state. The basic function of the retinol-binding protein (transfer and release of vitamin A) is tightly associated with the transition of this carrier protein into the molten globule like state, as in this case the release of retinol and protein denaturation are coupled and represent a common cooperative process. PMID- 8714357 TI - Extramural grant program. Chicago, Illinois, May 9-10, 1995. Abstracts. PMID- 8714358 TI - [The Hungarian nifedipines]. AB - The short history of the nifedipine drug substance containing Cordaflex product family--produced by EGIS Pharmaceuticals Ltd.--is presented. PMID- 8714359 TI - [A new synthesis of 1,4-dihydropyridines: cyclocondensation of aldimines]. AB - The reaction of 2-nitro-benzaldehyde with ammonia gives different products (1a-c) depending on the reaction conditions. All of them were suitable to prepare nifedipine, the well known Ca-antagonist compound. This new synthesis route is independent of the hitherto well-known methods. Moreover, the purity and stability of the received material prevails over the other marketed products. The reaction of 1a was studied with methyl acetoacetate and 3-aminocrotonate. PMID- 8714360 TI - [1H and 13C NMR investigation of nifedipine]. AB - 1H-13C shift-shift correlation based assignment of the 13C and 1H NMR spectra of nifedipin are presented along with some results of MM+ and PM3 conformation analysis. Both computation methods yield asymmetric low energy conformers. MM+ molecular dynamics simulations give similar results. A lower limit to the interconversion rate of the asymmetric low energy conformers is determined using magnetic shielding differences obtained by TNDO/2 calculations. PMID- 8714361 TI - [Photostability of nifedipine]. AB - Photochemical degradation reactions and the chemical structure of the decomposition products were studied in the 1,4-dihydropyridine series for Nifedipine drug substances and for various dosage of it (Cordaflex product family). The efficiency of the used primary and secondary light protection implemented for the Cordaflex retard 20 mg tablet were studied using 48-month long-term stability trials. The dissolution profiles and impurity levels (HPLC) obtained during the whole trial period met the strictest compendial requirements for this dosage form. PMID- 8714362 TI - [Investigation of the impurity profile of nifedipine by LC/MS]. AB - The manufacture-characteristic impurity profile of nifedipine drug substance was studied by HPLC and on-line LC/MS coupling. Using a new gradient HPLC procedure and MS detection, spectroscopic evidences for the chemical structure of some previously unknown minor impurities (all under 0.1%) were obtained. PMID- 8714363 TI - [Comparative human pharmacokinetic studies of 20 mg nifedipine-containing Cordaflex and Adalat film coated retared tablets]. AB - Comparative pharmacokinetic studies have been carried out with two 20 mg nifedipine active substance-containing retard film coated tablets, Cordaflex produced by EGIS Pharmaceuticals Co., Ltd. and Adalat of Bayer AG. The pharmacokinetic parameters and the relative bioavailability were determined in 15 and 16 healthy male volunteers, respectively after single and repeated administration in open, randomized cross over study. The plasma concentration of nifedipine was determined by HPLC-ED method, using laboratory robot for automated sample preparation. On the basis of graphical and statistical comparison of the pharmacokinetic parameters (AUC0-infinity, AUCss,0-tau, tmax, Cmax, Css,min, Css,av, MRT, etc.) calculated from the time-plasma concentration curve, moreover on the basis of clinical results, there was no significant difference between the two preparations. In conclusion, the relative bioavailability of Cordaflex and Adalat 20 mg retard tablets did not show significant difference after single and repeated administration. PMID- 8714364 TI - [Pharmaceutical technology of Cordaflex tablets]. AB - First the possibilities of solubilization and the photosensibility of nifedipine (the active ingredient of Cordaflex tablets) were investigated. The technology of retard tablet involves the preparation of a coprecipitate through spraying a solution containing nifedipine, a hydrotropic and a reardizing substance on carrier. After drying the produced granulated material was blended with common auxiliary ingredients, compressed into tablet and coated. The ratio of the two types of coprecipitating substances has a direct effect on the dissolution, so it proved predictable. The reproducibility of technology was good. PMID- 8714365 TI - [Selected passages from the clinical trials of Cordaflex]. AB - After a short review of the pharmacological characteristics of nifedipine this article presents a selection from the postmarketing clinical trials performed with Cordaflex (nifedipine) in Hungary. It summarizes the most favourable clinical effects and also the side effects observed with the different pharmaceutical formulations after short-term and long-term application. PMID- 8714366 TI - The role of growth factors in preimplantation development. AB - It has become clear that the mammalian embryo participates in a complex dialogue with the maternal physiology. The language of the dialogue is growth factor signalling. The embryo expresses receptors for insulin, IGFs, GH, EGF and cytokines including LIF, and CSFs; whilst ligands are secreted by the supporting tissues of the oviduct and uterus, and in some cases, the embryo itself. In the preimplantation period when the embryo is travelling to the uterus and passing through its first differentiation, these ligands affect embryonic physiology, apparently in ways that optimise developmental potential and synchronise embryonic and maternal physiologies. It is not yet clear in most cases whether this is by autocrine, paracrine or endocrine mode. In the crucial peri implantation phase the embryo is preparing to invade the maternal system for which extensive uterine remodelling is necessary. A model is proposed in which a cascade of growth factor activities, orchestrated by the ovarian steroid patterns, choreographs the biochemical players (ECM proteinases and their inhibitors) which initiate this activity. PMID- 8714367 TI - Biochemical and mitogenic properties of the heparin-binding growth factor HARP. AB - Heparin affin regulatory peptide (HARP), also called Pleiotrophin (PTN), is a polypeptide that displays a high affinity for heparin and that shares approximately 50% sequence homology with Midkine (MK). According to this structural homology, these two molecules constitute a new family of heparin binding proteins. The biological properties of HARP and MK remain largely a subject of debate. Both proteins have been described as neurite outgrowth promoting agents whereas until recently the mitogenic activity has been controversial. The aim of this review is to summarize the information on HARP with special focus on the recent data relating to its mitogenic properties. PMID- 8714369 TI - Bibliographic update. PMID- 8714368 TI - The role of fibroblast growth factor-2 (FGF-2) in hematopoiesis. AB - Basic fibroblast growth factor (bFGF or FGF-2) is an angiogenic and pleiotropic growth factor involved in the proliferation and differentiation of numerous cell types. It is expressed mostly in tissues of mesoderm and neuroectoderm origin, and is thought to play an important role in the mesoderm induction. Although hematopoietic cells derive from the mesoderm, relatively few studies have, until recently, addressed the role of FGF-2 in hematopoiesis. FGF-2 is expressed in cells of the bone marrow including stromal cells, and possibly cells from several hematopoietic cell lineages. It is stored in the bone marrow extra-cellular matrix and released by enzymes such as heparanase, plasmin, or phospholipase C and D. FGF-receptors (FGF-Rs) are expressed in leukemic cell lines and in hematopoietic cells. FGF-2 positively regulates hematopoiesis, by acting on stromal cells, on early and committed hematopoietic progenitors, and possibly on some mature blood cells. The action of FGF-2 is most likely indirect since its action, on megakaryocytopoiesis for example, is abrogated by anti-IL6 antibodies. It synergizes with hematopoietic cytokines, or antagonizes the negative regulatory effects of TGF-beta. Taken together, these results demonstrate that FGF-2 is a potent hematopoietic growth factor that is likely to play an important role in physiological and pathological hematopoiesis. PMID- 8714370 TI - [The maturation of intestinal digestive enzymes in ontogeny]. PMID- 8714371 TI - [The effect of stimulation of the suprachiasmatic and supraoptic nuclei of the anterior hypothalamus on the evoked activity of the visual cortex in rabbits]. AB - Stimulation of the hypothalamus' suprachiasmatic (SCH) and supraoptic (SO) nuclei induced short-latency hypothalamo-cortical responses in the visual cortex affecting the light-induced responses. Possible mechanisms of the differential effects of the stimulation upon different phases of the visual EPs, and participation of the RF in these mechanisms, are discussed. There seems to exist an interconnection between the hypothalamic and light stimuli in the visual cortex. PMID- 8714372 TI - [The effect of armin on neuromuscular transmission in the frog against a background of hexadecamethonium action]. AB - Hexadecamethonium as well as its alkylating derivatives significantly decrease the effect of organophosphorous acetylcholinesterase inhibitor armine on the amplitude and duration of the EPPs. It seems possible that these substances prevent inhibition of acetylcholinesterase. This property of the substances seems to be due to a polymethelene chain with two nitrogens as the basic part of the molecule structure. PMID- 8714373 TI - [An electrophysiological analysis of the processes in the evaluation of successfulness in the choice of behavioral programs]. AB - The magnitude of the N1 and P3 waves of event-related potentials (ERPs) recorded in the dog medial frontal cortex were shown to depend on the behavioural context of separate trials. The most prominent P3 waves occurred in two behavioural situations: 1) when an unexpected non-reward followed previously rewarded conditioned responses, and 2) when an absence of the reward resulted in behavioural type shift in the next trial. The enhancement of the P3 seems to be due to a common factor: the discordance between the CR or its results and the previously developed behavioural trend. PMID- 8714374 TI - [The characteristics of amino acid transport in the small intestine of chicks with avitaminosis A]. AB - An increased amino acid absorption occurred early in chicks in the vitamin A deficiency. This data suggests an effect of the vitamin A upon differentiation of amino acid channels within apical membrane of the absorbing cells. PMID- 8714375 TI - [The effect of serotonin on the receptors of the cat small intestine]. AB - 5-HT was shown to increase dose-dependently the afferent activity in the cat mesenteric nerves. Metergolin suppressed the effects of serotonin. The data obtained suggest that the small intestine afferents may take part in reception of immune response. PMID- 8714376 TI - [The effect of calcitrin and corticotropin on pulmonary surfactant and engorgement in rats during activation of the lateral hypothalamic area]. AB - The level of phospholipids, cholesterol, lipase activity in bronchoalveolar washes and lungs' blood capacity were studied in rats. An action exerted upon the hypothalamus led to a decrease in the surfactant lipids and the blood capacity of lungs. Possible peptidergic mechanisms of this are discussed. PMID- 8714377 TI - [The systemic hemodynamics of waking and anesthetized male cats during noradrenaline infusion]. AB - Dynamics of arterial pressure and its hemodynamic components were shoen to dramatically differ in corscious and anesthetised animals. The data obtained suggest that corscious male cats, along with a baroreflex mechanism of resistance against involvement of the heart into the pressor response, have another mechanism acting in deficiency of baroreflex effects. PMID- 8714378 TI - [The effect of epiphysectomy on the circadian dynamics of the cardiointervalogram indices of rats]. AB - Daily variations the heart rate variability were observed. Pinealectomy increased the sympathetic tone and shifted its acrophase to earlier hours in rats. PMID- 8714379 TI - [The dynamics of cardiac vagal sinus arrhythmia in cats under the action of met enkephalin]. AB - When stimulating the peripheral and of the right n. vagus in anesthetised cats, the sinus arrhythmia occurred in case the vagal bursts were beyond the limits of the controlled bradycardia range. I. v. administration of met-enkephaline suppressed the arrhythmia immediately but the latter reappeared within 15-20 minutes. The data obtained suggests a biphasic effect of met-enkephaline upon the vagal sinus arrhythmia. PMID- 8714380 TI - [The characteristics of the impulse activity of the motor units of the muscles in the pelvic floor in the cat]. AB - The investigation have been designed to elucidate the background and thermoregulatory activity (TA) of the motor units (MUs) of pelvic floor muscles (m. spincter ani externus, m. levator ani) and hind limb muscle (m. sartorius) in cats under anesthesia. The MUs of m. sphincter ani externus presented two patterns of background activity--the stationary and the rythmical ones. M. levator ani and m. sartorius presented no activity at room conditions (+22 degrees C). General cooling evoked TA in all the investigated muscles. The discharge frequencies of pelvic floor MUs (4-8 imp/s) during TA were shown to overlap with those of hind limb, muscles (5-9 imp/s). The correlation of MUs bioelectrical characteristics with histochemical and contractile properties of m. sphincter ani externus as fast-twitch muscle is discussed. The possible mechanisms responseble for MUs control in skeletal muscles with different specialization in modulated thermoreceptory afferent input are proposed. PMID- 8714381 TI - [The effect of polypeptides released by the thrombocytes during a liberation reaction on immunity and hemostasis]. AB - Low molecular peptides contained in the platelets manifested an anticoagulant and antifibrinolytic action, and affected the lymphocyte receptor expression. The polypeptides seem to take part in immune as well as hemostatic processes. PMID- 8714382 TI - [The effect of the duration of daylight on human hormonal and biochemical indices in the north]. AB - Change of the duration of the light day was shown to tell on biochemical and hormonal parameters in humans living on the North. The contents of T3 significantly increased in peripheral blood, the levels of insulin and urea decreased in March and October. Complex rearrangements of the metabolism in the course of year specific for high latitudes were revealed in humans. PMID- 8714383 TI - [The interrelation between the secretory function of the bovine udder and the amount of prolactin receptors on the membranes of the milk fat globules during lactogenesis]. AB - The prolactin receptors number reaches its maximal level in proestrus. The data obtained suggest a correlation between high-affinity prolactin receptors concentration on the milk fat globule membranes during proestrus and the cow milk yield. The membranes may provide a model for assessing the functional state and secretory activity of the cow mammary gland. PMID- 8714384 TI - [Epithelial-smooth muscle interaction in the regulation of the tonus of the airways in rats]. AB - The rat trachea epithelium was found to affect a catecholamine-induced relaxation of the trachea smooth muscles, to release a relaxing factor, the producing of which depends on calcium and is regulated by calmoduline and proteinkinase C. The epithelium-induced relaxation seems to depend on the animal age. PMID- 8714385 TI - [Seasonal changes in the functional state of the liver mitochondria in different animals]. AB - A difference in velocity of oxydation of the ATP substrates synthesis, conjugation and efficacy of oxydation of phosphorylation in animals with different functional systemic state, and seasonal differences in these processes, were found in different animals. PMID- 8714386 TI - [The effect of docosatrienoic acid on the potassium channels of outward rectification in the membrane of rat peritoneal macrophages]. AB - Application of docosatrienic acid was shown to dose-dependently decrease the peak K+ current amplitide and accelerate the potassium activation and inactivation kinetics at all membrane potentials. The data obtained suggests a direct effect of docosatrienic acid on the K+ channels in peritoneal macrophages. PMID- 8714387 TI - [The auditory perception of frequency-localized acoustic on-events]. AB - The quick changing of amplitude envelope (amplitude modulation) is the inherent properties of speech signals. They are analyzed by the auditory system and are reflected in the auditory sensation as a sequences of some "events", that participate in creating of phonetic images. The dynamics of frequency distribution of events is considered to be used by listeners as a determinants of some consonants. However, this supposition are inconsistent with the psychoacoustics experiments showed that listeners can't determine the frequency of amplitude irregularities of tonal components. Current work consists of the discussion of this problem and results of experiments where the perception of an amplitude jumps in different frequency format area was studied using the synthetic quasi--syllable. The results obtained give evidence that the phoneme interpretations "know" the frequency of amplitude irregularieties. PMID- 8714388 TI - [The perception of the tone and noise quality of a sound signal]. AB - Perception of tonalness and noisiness of tonal complexes with flat amplitude spectrum, was studied. The variable parameters involve: bandwidth, spacing of components, position on the frequency axis, and the depth of the amplitude modulation. The findings suggest that the tonalness and noisiness perception depends on a periodicity ad spectral processing. PMID- 8714389 TI - [Glycosaminoglycans of erythroblastic islands in the suppression and subsequent stimulation of erythropoiesis]. AB - The GAG types of the rat bone marrow erythroblastic islands were found to be similar to those of human macrophages. Activation of erythropoiesis with the erythropoietin induced an expression of heparin-sulfate. PMID- 8714391 TI - [Immunity--technical aspects of vaccines. I. Immunoprophylaxis in dogs and cats]. PMID- 8714390 TI - [Age-related changes in the activity of the peptidergic endings in the skin of rats]. PMID- 8714393 TI - [Vaccination failures. III. Postvaccinal complications and immunization schedules. Immunoprophylaxis in dogs and cats]. PMID- 8714392 TI - [Vaccination in practice. II. Immunoprophylaxis in dogs and cats]. PMID- 8714394 TI - [Current developments in the field of vaccines]. PMID- 8714395 TI - [ Veterinary drug reactions in aquaculture]. PMID- 8714396 TI - [Bovine spongiform encephalopathy (BSE). Examination in abattoirs compulsory as per March 1 1996]. PMID- 8714397 TI - Ukrainian National Collection of Microorganisms. AB - A list which describes microbial pools of the Ukrainian National Collection of Microorganisms created on the basis of the Institute of Microbiology and Virology of the National Academy of Sciences of Ukraine is presented. Collection guarantees maintenance of over 3500 strains of microscopic fungi, yeasts, coryne- and nocardioforms, streptomycetes, methane-oxidizing and methylotrophic bacteria, bacilli, phytopathogenic and nitrogen-fixing bacteria, gram-positive catalase positive aerobic cocci, pseudomonads, lactic-acid bacteria and mycoplasmas. Over 250 type strains and over 300 strains of microorganisms useful for medicine, economy and biotechnology are maintained in the Collection. PMID- 8714398 TI - [The use of immunomodulators to enhance postvaccinal immunity to experimental poliomyelitis]. AB - A possibility to use immunostimulator dibasole and methyluracil to increase [correction of decrease] the level of neutralising antibodies to polioviruses [correction of polyviruses] of types I, II, III in response to immunization by the alive poliomyelitis vaccine was studied in experiments on rabbits. The higher level of immune response when using the mentioned drugs, especially methyluracil, has been established; repeated immunization is needed to develop antibodies to poliovirus [correction of polyavirus] of type III. PMID- 8714399 TI - [The antibiotic properties of the phytotoxic metabolites of Botrytis cinerea Pers]. AB - Antibiotic properties of substances of a phytotoxic complex from Botrytis cinerea have been studied for a number of phytopathogenic bacteria, phytopathogenic and toxigenic fungi as well as saprophytic yeast strains. High fungistatic activity of preparations of phytotoxic metabolites (PTM) has been stated for Dendrodochium toxicum, Myrothecium verrucaria, M. roridum, Aspergillus fumigatus, Penicillium urticae, agents of heavy human and cattle mycotoxicoses. The studied representatives of phytopathogenic fusaria differed significantly in sensitivity to PTM: all the strains studied of the species from section Discolor (Fusarium gibbosum 54,624 and 51,463, F. graminearum 108,269, F. sambucinum 54,968, F. culmorum 54,951) were actively inhibited by the PTM concentrations studied; F. sporotrichiella 52,290 proved to be insensitive to them; the strains studied of the species from sections Elegans (F. oxysporum 55,715, F. moniliforme 54,262) and Martiella (F. javanicum 54,075, F. solani 54,719) possessed different sensitivity to PTM. The overwhelming majority of the yeast strains proved to be resistant to the PTM concentrations studied, except for the strains of Trichosporon cutaneum. In this connection the fact of high sensitivity of one of the strains of Kluyveromyces marxianus var. marxianus to B. cinerea metabolites arouses great interest. The strains of studied phytopathogenic bacteria (Erwinia carotovora subsp. carotovora 8,982 and Clavibacter michiganense 13) were sensitive to higher concentrations of PTM. PMID- 8714400 TI - [The effect of a combined preparation based on live microbial cultures from the genus Bacillus on the course of experimental infectious endometritis]. AB - A new model of experimental endometritis of rats, accessible for a wide use has been suggested. It gives a possibility to objectively estimate the effect of applied drugs on the course of the disease. The expressed treatment effect of the developed associated microbe preparation of Bacillus genus bacteria under staphylococcal endometritis was established in experiment on the basis of clinic observations, bacteriological and histological analysis. PMID- 8714401 TI - [New hydrogels of methylsilicic acid with metals and the in-vitro evaluation of their sorption activity in relation to rotaviruses]. AB - Processes of adsorption of rotaviruses of different strains by enterol sorbents on the basis of hydrogel of methylsilicic acid with bi- and trivalent ions of 3d metals (iron, cobalt, copper) fastened in their matrix have been studied. PMID- 8714402 TI - [Cytokines and nonviral infections]. AB - Cytokines (interferons, tumour necrosis factor and interleukin-1) with marked polyfunctional properties are described. They are synthesized by different cells of both immune and non-immune systems of organism and are able to stimulate, induce or inhibit various immunological and biological reactions and processes. The effect exerted by these cytokines on the course of bacterial infections is analyzed. The cytokines possess protective properties which are demonstrated in activation of neutrophils, intensification of bactericidal and phagocytic activity of macrophages and natural cells-killers. PMID- 8714403 TI - [Clinical importance of monophasic action potential registration in long QT syndrome]. AB - For more than 30 years, the monophasic action potential has been used as an experimental tool for the study of myocardial repolarisation. With recent improvements in catheter design, the utility of the tool as a means to identify the bases for ventricular arrhythmias in humans has been greatly improved. Abnormalities of repolarisation leading to ventricular arrhythmia formation can be identified and specific pharmacological therapies may be evaluated. The pathomechanism of major arrhythmias (ventricular tachycardia, ventricular fibrillation) occurring in long QT syndrome (LQTS) is not yet fully elucidated. The authors have recorded the monophasic action potentials (MAP) of the right ventricle in three patients with LQTS and with previous episodes of major ventricular arrhythmias. The changes in MAP duration and after depolarisation in response to spontaneous arrhythmias, programmed electrostimulation, atrial pacing and isoproterenol treatment were studied. In all of the three patients the early afterdepolarisation was present, which in two cases exhibited pause-dependent features. Thus in these patients dual chamber pacemaker implantatious were performed. These were the first permanent recordings of MAP during electrophysiological examination in Hungary and also the first evidences that the early afterdepolarisation does play a pathogenic role in the development of idiopathic LQTS. Further evaluation of the technique by cardiac electro physiologists may improve both the diagnosis and the treatment of ventricular arrhythmias dependent upon afterdepolarisation formation. PMID- 8714404 TI - [Endemic iodine deficiency in Hungary and possibilities of iodine substitution]. AB - Following a short historical review the authors summarize the latest results regarding domestic iodine supply. Iodine content in drinking waters, samples of mother's milk, urine and goiter screening data from all regions of Hungary unanimously verify that significant part of the population is deficient in iodine, yet supplementation of iodine is still even today an unsolved problem in our country. In the search for the reasons the authors review the theoretical possibilities of iodine supplementation by discussing advantages and disadvantages of the individual models. They establish that alimentary iodine supplementation in itself is not suitable due to both theoretical and practical reasons for the complete elimination of iodine deficiency, therefore they make recommendations to supplement iodine in other ways. They also discuss the effect of selenium deficiency on the metabolism of thyroid hormones. They call attention that domestic, endemic selenium deficiency may be related to the frequency of Hungarian occurrences of iodine deficiency disorders. The authors review the iodine deficiency disorders and their effects on the health of present and future generations. They compare the practical benefit and costs of iodine supplementation, and furthermore discuss in detail the possible side effects of iodine supplementation. PMID- 8714405 TI - [Endoscopic management of benign strictures of the common bile duct]. AB - 3 females with benign hepatic duct strictures were treated endoscopically. The treatment consisted of placement of one or two endoprostheses in all cases and endoscopic balloon dilatation in two cases. The stent was removed from the first patient--who had hepatic duct stones as well--after a 15 months stenting period and mechanical lithotripsy and balloon dilatation were performed with good results. The second and third patient-after stent insertion--was operated with suspicion of malignancy. Tumor was not proved. In the second patient the endoscopically placed stent was changed by a large diameter teflon stent surgically. The third patient's stents were left in place. These stents were removed after 3 months and balloon dilatation was performed. Patients' jaundice subsided and cholangitis improved following the stent insertion. COMPLICATIONS: Stent occlusion developed and common bile duct stones occurred once. There was not observed any other complication. Benign hepatic duct strictures can be treated by combination of prolonged endoscopic biliary stenting and balloon dilatation successfully. PMID- 8714406 TI - [Splenic metastases]. AB - Splenic metastases were found in 58 cases (3.1%) at autopsy out of 1842 widely disseminated solid tumors: melanomas, carcinomas of the lung those of the breast, uterus and liver being the most common primary tumors. This rare hematogenous metastatic spread could develop due to portal congestion. PMID- 8714407 TI - [A case of herpes simplex encephalitis requiring surgical management]. AB - The authors report about surgical treatment of herpes simplex encephalitis, although conservative management of this disease is the method of choice in the first place. They draw a lesson from this case that certain cases of large space occupying lesions of infectious origin leading to brain stem compression may require surgical management, even if they customary treatment is conservative. PMID- 8714408 TI - [From the history of the obstetric-gynecologic department of the Szent Margit Hospital in Hungary (1923-1963)]. PMID- 8714409 TI - [Relationship between physicians and art]. PMID- 8714410 TI - Partial identification of underdetermined compartmental models: a method based on positive linear Lyapunov functions. AB - A class of models is considered in which the number of unknown parameters exceeds the number of independent equations that are derived from the input/observation configuration. It is shown that, under certain circumstances, if the number of unknown parameters is sufficiently decreased by setting some of the exit parameters to zero, then the fallaciously derived model has the following interesting properties. The functions of time that give the amounts in the compartments are constant multiples of their counterparts in the fallacious model. A subset of the fallacious fractional transfer coefficients have the same values as their counterparts in the true model whereas others yield interval bounds. These and related results are obtained from a method based on positive linear Lyapunov functions. PMID- 8714411 TI - A stochastic model for circulatory transport in pharmacokinetics. AB - A new stochastic model for the residence time distribution of a drug injected instantaneously into the circulatory system is proposed and analyzed. The properties of the residence time are derived from the assumptions made about the cycle time distribution and the rule for elimination. This rule is given by the probability distribution of the number of cycles needed for elimination of a randomly selected molecule of the drug. Only the geometric distribution has been previously used for this purpose. Its transformation is applied here to get a boundary for the residence time. Other discrete distributions are applied with a view to describing different experimental situations. Suitable continuous probability distributions for the cycle time description are discussed. PMID- 8714412 TI - Population dynamic models generating the lognormal species abundance distribution. AB - This paper deals with a new class of stochastic species abundance models where the abundances are the points of an inhomogeneous Poisson process. These models are the result of a dynamic approach in which the changes in abundances through time are described by a multivariate diffusion and speciation constitutes a homogeneous Poisson process. In particular, the lognormal model is generated by assuming that the density regulation within each species is given by the Gompertz curve and that the environmental variances are constant. A substantial generalization is obtained by introducing a general type of interspecific density regulation and correlated environmental noise. This more general mechanism also generates the lognormal species abundance distribution. PMID- 8714413 TI - Carrier-borne epidemic models incorporating population mobility. AB - A study was made of multigroup epidemic models in which individuals are able to move between groups, infectious contact occurring only between an infective and a susceptible in the same group. Because of the mathematical intractability of such models; we look mainly at the number of susceptibles directly contacted by the infectives that are initially introduced into the population, ignoring subsequent infections caused by these newly infected individuals. We thus have a generalization of the carrier-borne epidemic model of Weiss [Biometrics 21:481 491 (1965)]. We consider first a model in which only infectives are able to move, then one in which both infectives and susceptibles move between groups. In each case we study both deterministic and stochastic versions of the model, concentrating mainly on the effect of varying the speed at which individuals move between groups on the number of initial susceptibles contacted. For the case in which only infectives move, the model is compared with a suitably matched model in which there is no movement between groups but infectives are able to infect outside their own group. The paper concludes with remarks on the behavior of the epidemic process if initially susceptible individuals that become infected are able to contribute to the further spread of the disease. PMID- 8714414 TI - Immunization levels for preventing epidemics in a community of households made up of individuals of various types. AB - A method is proposed for computing an epidemic threshold parameter for the spread of a communicable disease in a community of households in which individuals are of p different types. The threshold parameter is the largest eigenvalue of a p x p matrix whose elements depend on the rates of disease transmission between types and the distribution of the household size. More explicit expressions are given for diseases that are highly infectious within households, to the point that the infection of any member of a household results in the infection of all susceptible members of that household. For a variety of vaccination strategies it is described how this approach can be used to determine the level of immunity required to prevent epidemics. A numerical example illustrates the results. PMID- 8714415 TI - Hemotympanum. PMID- 8714416 TI - Endoscopic view of the recirculation phenomenon of sphenoid sinus drainage. PMID- 8714417 TI - Changing the nasal tip: Part III (of VI parts). PMID- 8714418 TI - Vocal fold paralysis. PMID- 8714419 TI - Sinus tympani: retrofacial approach for the removal of cholesteatomas. AB - The epithelium basement membrane of cholesteatomas in cases of chronic otitis media often extends into the sinus tympani. In the majority of cases it can be safely and completely removed by dissection through the ear canal and tympanum. In 2% of cases, not all squamous epithelium basement membrane can be removed because the sinus tympani extends more than 3 or 4 mm posterior to the anterior edge of the facial nerve and because the membrane is adherent to the irregular bony surface. Surgical exposure of the sinus tympani through the mastoid posterior and medial to the facial nerve allows complete removal of all basement membrane with a direct view. The technique using a retrofacial approach to expose this obscure sinus is described and a series of 48 cases treated successfully by this method are reported. PMID- 8714420 TI - To abandon or not abandon: antesigmoid craniotomy with retrolabyrinthine vestibular nerve section. AB - Increasing popularity of the retrosigmoid approach would have one believe this is the preferred posteriorfossa approach for a vestibular nerve section. The authors take issue with this. The antesigmoid posterior lateral retrolabyrinthine approach is a gentle craniotomy, easily performed and with low morbidity. A review of 70 consecutive cases revealed no procedural alteration because of surgical exposure, an absence of meningitis, no VIIth nerve weakness, minimal headache, average hospital stay of five days, and only one patient with a significant CSF leak. Hyperventilation anesthesia, patience during delicate VIIIth nerve exposure, incorporation of abdominal fat into dural closure sutures and prophylactic antibiotics have made this operation easy for the patient...and the surgeon. Don't abandon it! PMID- 8714421 TI - Kimura's disease (angiolymphoid hyperplasia with eosinophilia) of the tympanic membrane. PMID- 8714422 TI - Asthma: an important disease to otolaryngologists--Part I: Suspecting and diagnosing asthma. AB - Asthma is a chronic inflammatory disease of the lower respiratory tract which is triggered by exposure to allergens or other airway irritants. This inflammation results in airway hyperresponsiveness, bronchial muscle spasm, mucous gland hypersecretion and mucosal edema, which combine to create symptoms such as cough, wheezing and respiratory distress. Because the inflammatory process is highly variable, asthma is a disorder with many possible presentations. It may therefore proceed for years without clinical recognition, and may challenge the most astute diagnostician. It is important for otolaryngologists to be able to suspect, diagnose and treat asthma. This is so because asthma is a common disease in the otolaryngologic patient population, both as one of the options in the differential diagnosis of respiratory complaints, and as a comorbid condition which may complicate the treatment of other medical or surgical problems. Furthermore, both the understanding of asthma's pathophysiology, and its optimum treatment methods have undergone radical changes during the past decade. This three-part discussion reviews our modern understanding of asthma, and proposes diagnosis and treatment guidelines which can assist otolaryngologists in effectively managing their asthmatic patients. Part one summarizes current information on the pathophysiology and increasing prevalence of asthma, its clinical variability, the assessment of asthma sensitivity, and methods for diagnosis of asthma. Parts two and three cover the strategy for asthma management, and the use of both adjunctive and anti-inflammatory therapies for asthma control. PMID- 8714423 TI - Gelcap-assisted fiberoptic nasogastric feeding tube placement. PMID- 8714424 TI - Chondroid syringoma of the head and neck: clinical management and literature review. AB - This report describes the case of a chondroid syringoma occurring in the nasofacial groove of a 60-year-old woman. This benign, mixed epithelial tumor is infrequently seen by the otolaryngologist-head and neck surgeon, and therefore may not be included in the differential diagnosis of a nodular lesion on the skin of the head and neck. However, the most frequent site of occurrence for these relatively rare cutaneous lesions is the head and neck region. Histologically, these tumors are quite similar to pleomorphic adenomas of salivary gland origin, and optimal surgical management similarly requires removal of a cuff of normal tissue, rather than a simple shelling out of the lesion. The clinical presentation, pathology and recommended management of this rare tumor are discussed. PMID- 8714426 TI - Physical attractiveness of girls with gender identity disorder. AB - University students, masked to group status, judged the physical attractiveness of girls with gender identity disorder and clinical and normal control girls, whose photographs were taken at the time of assessment (mean age, 6.6 years). Each student made ratings for all girls for five traits: attractive, beautiful, cute, pretty, and ugly. A multivariate analysis of variance showed a significant group effect. Multiple comparisons of the significant univariate effects showed that the girls with gender identity disorder had significantly less attractive ratings than the normal control girls for the traits attractive, beautiful, and pretty who, in turn, had less attractive ratings than the clinical control girls. Girls with gender identity disorder and the normal controls also had less attractive ratings than the clinical controls for the trait cute. Correlational analyses showed that age was substantially negatively related to the attractiveness ratings in the group of girls with gender identity disorder, but was considerably less so in the two control groups. The extent to which the group differences in attractiveness were due to objective, structural differences in facial attractiveness vs. socially created, or subjective, processes is discussed. PMID- 8714425 TI - Effects of yohimbine on sexual experiences and nocturnal penile tumescence and rigidity in erectile dysfunction. AB - The therapeutic effect of the alpha 2-antagonist yohimbine in erectile dysfunction was studied in a double-blind placebo-controlled design. Thirty-one male patients underwent extensive clinical, urological, and psychiatric diagnosis and were dichotomically classified into an organic and a nonorganic subgroup. Following a 1-week placebo run-in period, patients were randomly assigned to a placebo or a verum group (yohimbine 15 mg daily) for a treatment period of 7 weeks. The Clinical Global Impression (CGI) scale was used as the primary efficacy parameter. Additionally, nocturnal penile tumescence and rigidity (NPTR) were measured. Global assessment of erectile function applying the CGI scale revealed, beyond a placebo effect in both organic and nonorganic patients, a therapeutic effect in the subgroup of nonorganic patients, with a significantly greater improvement in the yohimbine group compared to the placebo group. No superiority of yohimbine compared to placebo was found in the organic patients. These findings on the subjective level had no correlate in the NPTR recordings. The NPTR parameters were unchanged under yohimbine treatment in both the nonorganic and organic subgroup. No interrelation was found between subjective improvement and NPTR alterations. Polysomnographic control of the NPTR registrations ensured that the duration of REM sleep under treatment was not influenced. PMID- 8714427 TI - Ethnic and gender differences in sexuality: variations in sexual behavior between Asian and non-Asian university students. AB - Seven hundred and two (346 non-Asian, 356 Asian) undergraduate volunteers were assessed in a confidential laboratory setting on levels of interpersonal sexual behavior (e.g., petting, intercourse), intrapersonal sexual behavior (e.g., fantasy, masturbation), and sociosexual restrictiveness (e.g., lifetime number of partners, number of "one-night stands"). The purpose was to examine possible differences in sexual behavior between Asian and non-Asian Canadian university students and to determine the association between North American residency and the sexual behavior of Asians. The role of gender on sexual behavior both across and within ethnic groups was also examined. Statistical analyses revealed that Asian students were significantly more conservative than non-Asian students on all measures of interpersonal sexual behavior and sociosexual restrictiveness. Significant differences were also noted between Asian and non-Asian students on most measures of intrapersonal sexual behavior. With the exception of two fantasy items, length of residency in Canada was unrelated to interpersonal sexual behavior, intrapersonal sexual behavior, or sociosexual restrictiveness among Asians. Although gender differences were substantial for intrapersonal sexual behaviors such as fantasy and masturbation, no significant gender differences were found for measures of interpersonal sexual experience, with the exception of reported number of one-night stands. PMID- 8714429 TI - Gender differences in the organization of sexual information. AB - One widely used model of knowledge representation is that of a network in which concepts are portrayed as nodes with links between the nodes representing associations. Schvaneveldt (1990) developed a method (Pathfinder) that generates associative networks from individual's ratings of similarity of word pairs. We had 51 female and 47 male undergraduates rate for similarity all paired combinations of 16 words judged as relevant to the domain of sexuality. Using a measure of network similarity, we found that each gender's networks were more similar to each other than they were to the other gender's. Using the number of links on words as the dependent variable there were gender differences in the number of links within clusters of words, between clusters of words, and on specific words. These differences, for the most part, are consistent with gender stereotypes and prior research showing gender differences in the processing of sexual information. PMID- 8714428 TI - Oral contraceptives and sexuality in university women. AB - The relationship between use of oral contraceptives and sexuality in university women who completed the McCoy Female Sexuality Questionnaire was examined. Pill users reported sexual intercourse earlier, were less likely to be virgins, more likely to have a sex partner, more apt to be engaging in petting and intercourse than nonusers, and reported more frequent intercourse than sexually active nonusers. Nonusers reported a greater frequency and enjoyment of anal intercourse than pill users. The prediction that pill users would have less vaginal lubrication than nonusers was supported. Contrary to prediction, pill users reported a higher frequency of sexual thoughts and fantasies, and level of sexual interest than active nonusers. Triphasic Orthonovum 7/7/7 (OR7/7/7) users reported more sexual thoughts and fantasies and had higher Sexual Interest (Factor 1) scores than monophasic Orthonovum 1/35 (OR1/35) users. Triphasic users as a group enjoyed sexual activity more, were more aroused during sexual activity, and had higher Sexual Interest factor scores than monophasic users. Triphasic users reported more sexual thoughts and fantasies, sexual interest and had higher Sexual Interest factor scores than nonusers, while monophasic users did not. Monophasic users reported less vaginal lubrication than nonusers, whereas triphasic users did not. Comparisons of OR7/7/7 and OR1/35 users with nonusers revealed the same findings. Results suggest that women using triphasic pills experience greater sexual interest and response than those using monophasics. Possible reasons for these differences are discussed. PMID- 8714430 TI - A strategy to promote the rational use of laboratory tests. International Federation of Clinical Chemistry. AB - These guidelines suggest how clinical chemists may promote rational laboratory use by critically evaluating the tests they offer. This may be done by documenting the clinical uses and limitations of tests in the same way that properly written up method descriptions outline analytical procedures in a particular laboratory. This information can then be promoted on result report forms, in discussion or through investigational protocols or clinical guidelines. For these to be of value they should be the result of clinical and laboratory input reflecting the local practice and facilities. They should be clearly written, easily accessible to the users and updated whenever there is a change of methodology or clinical practice. Much can be achieved by applying common sense to common knowledge and laboratories of all sizes can prepare their own material. However, national scientific societies and other professional organisations can assist local endeavour by avoiding and disseminating educational material on test evaluation, selection and use. PMID- 8714431 TI - A one-step sandwich enzyme immunoassay for human matrix metalloproteinase 8 (neutrophil collagenase) using monoclonal antibodies. AB - A one-step sandwich enzyme immunoassay (EIA) system for human matrix metalloproteinase 8 (MMP-8, neutrophil collagenase, EC 3.4.24.7) has been established with a pair of monoclonal antibodies prepared against the zymogen of MMP-8 purified from human neutrophils. MMP-8 in samples simultaneously reacted with both solid-phase and peroxidase-labeled antibodies. Sensitivity of this EIA system was 0.34 micrograms/l (5.7 pg/assay) and linearity was obtained between 0.5 and 500 micrograms/l (8.3-8300 pg/assay). The EIA system recognized both precursor and active forms of MMP-8 but not MMP-8 complexed with tissue inhibitors of metalloproteinases. There was no difference in the MMP-8 levels between the plasma samples from patients with rheumatoid arthritis or osteoarthritis and those from healthy subjects (median 6.2 micrograms/l, range 1.5-28 micrograms/l). However, the level in synovial fluids from patients with rheumatoid arthritis (median 345 micrograms/l, range 84-2860 micrograms/l) was shown to be higher than that from osteoarthritic patients. MMP-8 levels in human whole saliva from patients with periodontal diseases (median 282 micrograms/l, range 0-1420 micrograms/l) were also significantly higher than those from clinically healthy subjects (median 25 micrograms/l, range 0-100 micrograms/l). Immunoreactivity analyses showed that MMP-8 species in normal human plasma exists as a precursor but not as a complex form with tissue inhibitor of metalloproteinases (TIMP)-1 or TIMP-2. PMID- 8714432 TI - Use of beta-maltosides (p-nitrophenyl-beta-D-maltoside, 2-chloro-4- nitrophenyl beta-D-maltoside and 4-methylumbelliferyl-beta-D-maltoside) as substrates for the assay of neutral alpha-glucosidase from human kidney and urine. AB - The method of assay of neutral alpha-glucosidase from human kidney and urine using beta-maltosides (p-nitrophenyl-beta-D-maltoside [NP-beta-D-maltoside], 2 chloro-4-nitrophenyl-beta-D-maltoside]) [CNP-beta-D-maltoside] and 4 methylumbelliferyl-beta-D-maltosides ([MU-beta-D-maltoside]) as substrates and beta-glucosidase as an auxiliary enzyme is proposed. All three beta-maltosides are suitable substrates for the determination of neutral alpha-glucosidase activity but MU-beta-D-maltoside is the most sensitive due to its methylumbelliferyl moiety. The method is simple, convenient and 10-fold more sensitive than the commonly used alpha-glucosidase assay procedure with the corresponding synthetic alpha-glucosides, p-nitrophenyl- alpha-D-glucoside (NP alpha-D-glucoside) and 4-methylumbelliferyl-alpha-D-glucoside (MU-alpha-D glucoside). A modification of the method, with p-nitrophenyl-beta-D-maltoside as substrate, was applied to the semiautomatic assay of urinary alpha-glucosidase in 96-well microtitre plates. PMID- 8714433 TI - Evaluation of endotoxiuria for diagnosis of urinary tract infection after major surgical procedures. AB - In 148 patients after major surgical procedures urinary endotoxin levels were determined and compared with bacteriological results. The study was designed as a screening study. Urine samples were collected once by suprapubic or transurethral catheters. In a first series of 49 patients urine bacteriology was positive (mainly, Gram-negative rods were found) in 3 cases. However, endotoxin determination was positive in these 3 patients and in a further 10 patients receiving antibiotic therapy for other reasons. Therefore, the following 99 patients were studied also by urinalysis by reagent strips for leukocytes and nitrite. In the second series, 12 urine cultures positive for bacteria were observed. Eleven samples were also endotoxin positive. Five more patients were endotoxin positive and had pathological but unspecific reagent strip results. These patients were treated with antibiotics for other reasons. Patients with candida found in the urine culture (n = 5) were endotoxin negative. Thus, endotoxin determination in urine obtained by suprapubic or transurethral catheters proved to be a very sensitive method for diagnosis of bacterial contamination, even during antibiotic treatment. PMID- 8714434 TI - Plasma parathyroid hormone related-protein levels in patients with cancer, normocalcemic and hypercalcemic. AB - Hypercalcemia in patients with cancer may reflect the synthesis and secretion into circulation of parathyroid hormone-related protein (PTHrP) produced by the tumor. In the present study, we have measured circulating PTHrP concentrations in healthy subjects and patients using a new immunoradiometric assay (IRMA) that is specific for the 1-86 amino acid sequence of molecule, and in plasma collected with protease inhibitors. Plasma concentrations of PTHrP(1-86) were greater than the detection limit of the assay (0.3 pmol/l) in healthy subjects. All patients with hypercalcemia-associated cancer had PTHrP(1-86) levels significantly greater (median 7.74 pmol/l, P < 0.05) than healthy subjects or patients with cancer and normal serum calcium, primary hyperparathyroidism and hyperparathyroidism secondary to chronic renal failure. Plasma PTHrP and corrected serum calcium were correlated in patients with hypercalcemia-associated cancer. In one patient, a marked decrease in PTHrP and calcium levels was observed following surgery. Our results suggest that this IRMA for PTHrP(1-86) may be useful for diagnosis and monitoring of PTHrP-producing tumors induced hypercalcemia. PMID- 8714435 TI - Investigation of erythrocyte membrane lipid peroxidation and antioxidant defense systems of patients with coronary artery disease (CAD) documented by angiography. AB - In the present study, erythrocyte membrane lipid peroxidation, erythrocyte superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and serum vitamin C levels of subjects with and without coronary artery disease (CAD) documented by coronary angiography have been determined. Patients consisted of 42 subjects (31 male, 11 female) aged between 41 and 77 years and controls consisted of 35 subjects (21 male, 14 female) aged between 40 and 69 years. Erythrocyte lipid peroxidation of the patients was significantly higher (P < 0.005) whereas erythrocyte SOD, GSH-Px activities and serum vitamin C levels were lower than those of controls. Our results show significant alteration in erythrocyte membranes and antioxidant status of patients with CAD. However, whether such alterations are a primary cause or consequence of CAD and whether these alterations can be used in the diagnosis and management of the disease needs further investigation. PMID- 8714436 TI - A one-step sandwich enzyme immunoassay for human matrix metalloproteinase 7 (matrilysin) using monoclonal antibodies. AB - A one-step sandwich enzyme immunoassay (EIA) system for human matrix metalloproteinase 7 (MMP-7, matrilysin, EC 3.4.24.23) was established with a pair of monoclonal antibodies prepared against the zymogen of MMP-7 (proMMP-7) purified from CaR-1 human rectal carcinoma cells and against an oligopeptide corresponding to the C-terminal domain of human proMMP-7. ProMMP-7 in samples was allowed to simultaneously react with both solid-phase and peroxidase-labeled antibodies. Sensitivity of the EIA system in the presence of EDTA was 0.05 microgram/l (1.5 pg/assay) and linearity was obtained between 0.156-10 micrograms/l (4.68-300 pg/assay). ProMMP-7 levels in human sera from healthy subjects were shown to be in the range of 10.7 +/- 18.8 micrograms/l. Gel filtration analyses indicated that proMMP-7 makes polymers after treatment with EDTA and suggested that proMMP-7 exists as polymers in normal human sera. PMID- 8714437 TI - Increased serum levels of neuron-specific enolase in epileptic patients and after electroconvulsive therapy--a preliminary report. AB - Serum neuron-specific enolase (NSE) levels were studied by an enzymo-immunoassay method in 2 groups of patients: a group of epileptic patients, and a group of patients with refractory major depression after electroconvulsive therapy (ECT). In patients without organic neurological disease (n = 274) the mean serum NSE level (+/- S.D.) was 8.4 +/- 3.4 micrograms/l. No correlation with sex or age was observed. No significant difference was observed between epileptic patients without seizure or major electroencephalogram (EEG) abnormality, and a reference group. Significant increases were observed in 32 samples collected from patients with interictal EEG without spikes and waves before the 7th day after a seizure, in whom mean NSE was 21.5 +/- 9.4 micrograms/l, and in 26 samples from 4 patients without seizures but with spikes and waves in the interictal EEG, whose mean NSE was 20.6 +/- 11.5 micrograms/l. The increases of serum NSE levels in epileptic patients seem therefore to be linked to seizures and/or to EEG abnormalities. The consequences of these observations for the survey of epileptic patients, and for the diagnosis of cerebral tumors (mainly neuroblastoma) or for monitoring treatment after surgical resection, are discussed. In only 1 patient out of 6, an increase in serum NSE levels was observed with a peak about 12 h after ECT. No significant correlation with the ECT features (length of seizures, one- or two sided electrodes) was observed. PMID- 8714438 TI - Radioimmunoassay of free thyroxine (T4) using 125I-labeled T4-IgG complex with very large molecular weight. AB - We describe a double-antibody radioimmunoassay for the determination of free thyroxine (FT4) in serum using a 125I-labeled T4-IgG conjugate of very large molecular weight (approximately 1,000,000) as tracer. The binding of this conjugate to the anti-T4 antibody was found to be satisfactory and independent of the T4-binding protein concentration. The T4 sequestration by the antibody was limited and the sample dilution used in the final assay mixture was low. The results determined by the assay developed for samples from hypo-, hyper-, euthyroid subjects and from non-thyroidal-illness patients were well correlated with those obtained by a free T4-equilibrium dialysis kit (Nichols). The detection limit of the assay was 0.41 pmol/l and the range covered by assay standard solutions was 2.4-121 pmol/l. The FT4 concentrations in serum as determined by this assay were 9.6-31 pmol/l (mean 21 pmol/l) for 46 euthyroid serum samples; 6.7-47 pmol/l (mean 23.5 pmol/l) for 27 non-thyroidal illness; and 4.2-25.8 pmol/l (mean 11.6 pmol/l) for 19 pregnant women. PMID- 8714440 TI - On the stability of human lysosomal enzymes at room temperature in normal and acidified plasma and serum. PMID- 8714439 TI - Leukocyte lipid peroxidation, superoxide dismutase, glutathione peroxidase and serum and leukocyte vitamin C levels of patients with type II diabetes mellitus. AB - In the present study, leukocyte lipid peroxidation, superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and serum and leukocyte vitamin C levels of patients with type II diabetes mellitus and healthy controls were investigated. Patients consisted of 53 cases (23 male, 30 female) aged 35-75 years and controls of 34 subjects (15 male, 19 female) aged 34-66 years. Leukocyte lipid peroxidation of diabetics was significantly increased (P < 0.05) whereas vitamin C level was decreased (P < 0.05) compared to those of controls. There was no significant difference in the other parameters. Also, there was no correlation between the above parameters and HbA1c and glucose levels. Our results show that leukocytes of diabetics are affected by oxidative stress which might be a reason for decreased microbicidal activity. PMID- 8714441 TI - Demonstration of a biliary-associated glycoprotein in human serum. PMID- 8714442 TI - Motivation and gambling involvement. AB - It is argued that motivation is a key determinant of gambling involvement. The hypothesis--that participants who exhibit a high self-determined motivational profile (i.e., engage in gambling for fun and have a sense of choice) will report a higher degree of involvement--was supported in a study conducted in Canada, with 245 gamblers who bet on horses. Also, the male participants were more involved in gambling than the female participants were. PMID- 8714443 TI - The motivation of Australian adolescent schoolchildren to engage in group discussions about bullying. PMID- 8714444 TI - [Latency times and preadmission procedures]. AB - During focal cerebral ischemia, irreversible changes occur at the core of the lesion, whereas energy metabolism and ion homeostasis are preserved in the surrounding penumbra. Not all brain cells are killed at the start of a stroke; some are destroyed hours or days later, usually by toxins released during the early stages. Time is therefore the critical factor in treating acute ischemic stroke. Early management by the neurologist is associated to better functional outcome and shorter hospital stays. Patient evaluation and treatment must therefore take place as soon as possible. PMID- 8714445 TI - [Emergency assessment in the hospital: the ability of early diagnostic tests to predict short-term prognosis, progression of infarct, hemorrhagic conversion and cerebral edema]. AB - Acute stroke should be assessed by a specialist on an emergency basis. This article reviews the ability of clinical data and the results of diagnostic tests performed within 6 hours of onset to predict short-term prognosis, neurological deterioration, hemorrhagic transformation of the infarct and cerebral edema. Scores on neurological deficit scales, early signs of infarct on cranial computerized tomography, biochemical parameters such as glycemia, and the characteristics of vascular occlusion on Doppler ultrasonography or angiography are good predictors of acute phase complications and can be used to help identify patients that might benefit from new stroke therapies. PMID- 8714446 TI - [Treatment of medical complications: hypertension/hypotension, hyperglycemia, hyperthermia, hypoxia, cardiac complications and pulmonary embolism]. AB - The stroke patient often suffers additional medical complications related to age, heart and artery damage and immobility. Pulmonary embolism is one condition that can sometimes be prevented. Heart disease must be detected and treated, as it is the principal cause of death after stroke. Recent data describing the physiology of stroke suggest that hypotension, hyperglycemia and hyperthermia should be prevented, as these conditions increase the likelihood of death or sequelae. Stroke is no longer a "wait and see" disease. PMID- 8714447 TI - [Treatment of neurological complications: cerebral edema and epileptic seizures]. AB - Acute ischemic stroke is a true medical emergency, as early diagnosis and treatment is the only viable way to open the so-called "therapeutic window" and prevent devastating consequences. Although neurological complications are few at onset and in the early hours, they are potential causes of death and morbidity. The main cause of death in the acute phase is cerebral edema. Early diagnosis and treatment, mainly with hyperosmolar agents, hyperventilation and/or surgery, can save lives and reduce the impact of sequelae. Early epileptic seizures are seen in 4-5% of stroke patients. Treatment is only needed in status epilepticus or recurrent episodes. We review the most important features and current therapeutic strategies for the management of complications. PMID- 8714448 TI - [Neuroprotection in cerebral ischemia]. AB - It have been reviewed the different mechanisms involved in the development of cerebral ischemia and infarct, as well as the possible interaction and potentiation among them. Experimental and clinical trials undertaken with different drugs, such as glutamate antagonists, calcium channel blockers and free radical antagonists are discussed on the light of their possible application in prophylactic neuroprotection. PMID- 8714449 TI - Thrombolysis in acute stroke. AB - The use of fibrinolytic agents in acute ischemic stroke has received increasing attention due to the completion of several prospective studies examining the efficacy and safety of these drugs in this clinical setting. Experience with plasminogen activators indicates that recanalization of carotid and vertebrobasilar territory occlusion is feasible within 4-6 hours of symptom onset. The optimal plasminogen activator, its dose-rate and delivery system, however, is not known. The phase III trial of the European Cooperative Acute Stroke Study (ECASS) suggests potential modest benefit in outcome, although recombinant tissue plasminogen activator (rt-PA) had an increased risk of hemorrhage attributable to those patients with early CT-scan signs of ischemia. Complete cervical internal carotid artery occlusions by in situ thrombosis appear more resistant to thrombolysis than occlusions of the stem and major branches of the middle cerebral artery. The issue of hemorrhagic transformation is unsettled. It seams to be increased by delayed intervention on the one hand, but it is not different from that observed in placebo patients in phase I/II series. PMID- 8714450 TI - [Antithrombotic treatment in acute ischemia stroke]. AB - Antithrombotic drugs (especially heparin and aspirin) are widely used to treat patients during the acute phase of ischemic stroke, yet there is no evidence of any beneficial effect on the course of the disease. Although these drugs can reduce the risk of such complications as deep venous thrombosis and, possibly, the number of recurrences of stroke, they can also give rise to systemic or cerebral hemorrhage. Given the uncertainty about the efficacy and safety of antithrombotic drugs, and specifically about the risk/benefit ratio, large clinical trials are needed to determine the indications for their use in the management of acute ischemic stroke. We stress the importance, because of its magnitude, of the International Stroke Trial (IST), the results of which will be available in 1996. PMID- 8714451 TI - [Early therapeutic intervention: thromboendarterectomy and percutaneous transluminal angioplasty]. AB - Occlusion of the internal carotid artery (ICA) due to atherosclerosis, or more rarely thromboembolism, is a fairly frequent cause of stroke. Spontaneous ICA occlusion is associated with a 40 to 69% incidence of profound neurologic deficit and a 16 to 55% death rate. Management of this entity, however, continues to be controversial, as currently used treatments have not yet been shown to modify morbidity/morbility rate. Acute-phase performance of thromboendarterectomy or percutaneous transluminal angioplasty may play a role in ICA treatment. Emergency revascularization has not generally been considered to benefit stroke patients: clinical improvement is only slight in many cases and the incidence of perioperative complications, particularly brain hemorrhage, is high. Review of the literature, however, demonstrates that adequate restoration of flow is achievable in a subgroup of patients with spontaneously occluded symptomatic ICAs without severe neurological deficit, without decreased level of consciousness and without intracerebral hemorrhage manifest in computed tomography or magnetic resonance images. The outcome of emergency intervention also seems to depend on the duration of symptoms, individual collateral blood flow and the angiographic reflux pattern in the ICA. Given the potential usefulness of these surgical techniques, controlled clinical comparison of operated and non-operated groups seems highly recommendable in spite of the significant difficulties involved. PMID- 8714453 TI - Age-related changes in the tip-of-the-tongue experience. AB - The tip-of-the-tongue (TOT) experience was compared in younger and older adults, using picture and definition cues to elicit TOTs. Older adults (a) experienced more TOTs, (b) had less target word information available during the TOT, and (c) had more related words (blockers) come to mind during the TOT. There were no age differences in either the speed or likelihood of immediate resolution of the TOT. Picture and definition cues were equally likely to evoke TOTs, although picture cues elicited more interlopers. The idea that TOTs are more likely to be elicited by less common target words was not supported. PMID- 8714454 TI - Effect of facial familiarity and task requirement on electrodermal activity. AB - We examined the effect of facial familiarity and task requirement on electrodermal activity (EDA). Proposed models of facial recognition suggest a sequential process wherein a recognition of familiarity precedes any identity specific search. Prior research has indicated that an automatic increase in EDA occurs to familiar faces. We reexamined this effect while manipulating the task requirement. One group of subjects was required to identify (name) faces, and a control group was required to rate facial attractiveness. The results indicated that an increase in EDA to familiar faces occurred only when coupled with the identification task. No increase in EDA occurred when subjects were rating facial attractiveness and presented with a familiar face. PMID- 8714455 TI - [The mechanism of muscle contraction: along the path carved by Academician G.M Frank]. AB - On the issue of the mechanism of muscle contraction, the views of the late G.M. Frank, to whom this symposium is dedicated, differed fundamentally from those of the then-current orthodoxy. Frank could not accept the idea of filament sliding brought about by swinging cross-bridges. He seemed particularly moved by two observations from his laboratory: the first, that the A-band shortened during active contraction; and the second, that sarcomere shortening occurred in stepwise fashion. From these findings principally, Frank came to the view that contraction involved a stepwise shortening of thick filaments. Although this view is broadly held to be incorrect, I will present evidence that it may constitute at least a partial answer. I will consider the evidence that both of Frank's principal findings-stepwise shortening and thick filament shortening-may be valid attributes of the contractile mechanism. I will then go on to show how these two attributes fit into an overall mechanism-not dissimilar to the one put forth by Frank. PMID- 8714456 TI - [Statistics on actin globule chains, stretched by hydrogen bonds, and the Hill law in quantum mechanics theory of muscle contraction]. AB - The process of sarcomere membranes approaching under the action of actin and myosin filament stretching is studied theoretically. In elaboration of previous studies it is supposed that this stretching is due to the formation of hydrogen bonds excited by the energy of ATP splitting. A possibility for the hydrogen bond breakage at different stages of contraction is considered to be the greater the more are the polymers stretched, and the larger statistical scatter in the moments of H-bonds formation at different filaments if it is supposed that sarcomere contraction occurs much slower than stretching of each separate filament. As a result, one obtains a relationship between the rate of contraction and the external force which is close to the Hill's law, and the external force efficiency of a muscle contraction and heat extraction value are related to parameters of polymers and their properties. PMID- 8714457 TI - [Some new perspectives on V. I. Deshcherevskiy's kinetic theory of muscle contraction]. AB - Deshcherevsky's kinetic theory of muscle contraction (Deshcherevsky, 1968, 1971) is considered in the light of the modern experimental data. The data are discussed, which support the validity of some propositions and conclusions of the theory, including the postulate on the negligibly small probability of pulling cross-bridge detachment. It is shown that if this postulate is accepted and the distance between the neighbouring actin sites for the given myosin cross-bridge attachment on the actin filament is taken as an actin filament half-turn (Sydorenko, 1982, 1984), then simple explanations, which don't contradict the cross-bridge hypothesis of force generation, could be given to a number of muscle behaviour features such as the high efficiency, stepwise shortening, repeated tension recovery after repeated forced step-wise shortening of activated muscle with the time-interval less than 10 ms, step-size up to 90nm powered by a single ATP molecule and so on. PMID- 8714458 TI - [Structure and functional significance of the thick filament]. AB - New models of the thick filament structure and muscle contraction are proposed in the work. Association of myosin molecules into filament occurs by intertwisting between them. The model is in good consistency with experimental data. A computer analysis of the widely accepted concept of parallel packing of myosin molecules is presented. It is shown that the rowing movement of crossbridges resulting from parallel packing is, on the one hand, impossible in terms of sarcomere construction, and on the other hand, is in conflict with available experimental data. The crossbridge movement by unwrapping easily agrees with recent experimental data as well as given possibility to interpret such phenomena as the change of the myofilament dimensions under certain conditions or step-wise manner of sarcomere contraction. PMID- 8714459 TI - [Correlation between Ca2+-dependent movement of crosslinks in myosin filaments and Ca2+-sensitive actin-activated ATPase of skeletal muscle myosin]. AB - The dependence of actin-activated ATPase activity and myosin filament structure have been studied on Ca(2+)-concentration in the range between pCa 7.5 and pCa 4.6. Rabbit skeletal muscle myosin with dephosphorylated regulatory light chains column-purified with DEAE-Sephadex A-50 for the removal of minor proteins and actin without regulatory proteins have been used. Considerable increase in actomyosin ATPase activity (by 70%) is revealed with increasing Ca(2+)-level from pCa 7.5 up to pCa 4.6. Electron microscopic observations on the structures of reconstituted myosin filaments have revealed Ca(2+)-dependent movement of myosin cross-bridges (head + subfragment-2) from and to the backbone of myosin filaments. The correlation between the manifestation of Ca(2+)-sensitivity of ATPase properties of myosins and Ca(2+)-dependent mobility of cross-bridges has been established. In particular, the increase in the mobility of cross-bridges and their moving away from the surface of myosin filaments at pCa 4.6 correlates with the increase in actin-activated ATPase of the same myosin preparations. It is supposed that the interrelation between the above properties observed in in vitro system can be of importance for force generation and its regulation in muscle. PMID- 8714461 TI - [Structural changes in actin filaments during binding with phosphofructokinase (F protein), detected using an optical diffraction method]. AB - It has been previously shown by us that phosphofructokinase (F-protein) binds to rabbit skeletal muscle F-actin and reconstituted thin filaments forming ordered bundles. Upon low molar ratios of phosphofructokinase to actin in the complex bundles the enzyme molecules are arranged between actin filaments regularly in the form of crossbridges. The increase of phosphofructokinase: actin ratio up to equimolar one and more leads to the filling of the space between actin filaments with phosphofructokinase-molecules. In these bundles the inclined cross striation with axial repeat of about 7.0 nm is clearly seen. Optical diffraction analysis of the micrographs revealed new features in the structure of such complexes in comparison with F-actin and reconstituted thin filaments. Optical diffraction patterns from F-actin and reconstituted thin filaments exhibit the first (35.5 nm) and sixth (5.9 nm) layer lines typical of F-actin helix structure. On the optical diffraction patterns from the complex bundles the meridional reflection of about 7.2nm is present, that is not observed on the optical diffraction patterns from F-actin alone. This reflection is characteristic of optical diffraction- and X-ray patterns of myosin filaments and is the sixth order of axial period of their structure (42.9 nm/6). The structural changes occurring upon binding is supposed to be important for the mutual regulation of functional activity of enzymes and actin-containing filaments in muscle. PMID- 8714460 TI - [Calorimetric study of stable complexes of myosin subfragment I with adenosine diphosphate and phosphate analogs]. AB - This short review is concerned with the application of the method of differential scanning calorimetry to study the conformational changes of isolated myosin head (myosin subfragment 1, S1) caused by the formation of the S1 complexes with Mg(2+)-ADP and P(i) analogues such as orthovanadate (V), aluminium fluoride (AIF4 ) or beryllium fluoride (BeFx). These changes of the whole S1 molecule are reflected in a significant increase of S1 thermal stability and in a pronounced increase of the cooperativity of the thermal denaturation. Since the complexes S1 ADP-V, S1-ADP-AIF4- and S1-ADP-BeFx are stable analogues of the S1**-ADP-P(i) transition state of the S1-catalyzed ATP hydrolysis, it is concluded that DSC studies with these complexes offer a new and promising approach to investigate the structural changes which occur in the myosin head during Mg(2+)-ATPase reaction. PMID- 8714462 TI - [The role of histidine residues in conformational changes in the sarcoplasmic reticulum Ca-ATPase active site]. AB - Conformational pH-induced changes of Mg-ATP binding site of the sarcoplasmic reticulum Ca-ATPase (SR-ATPase) were investigated by fluorescence energy transfer between covalently bound fluorescent label (fluorescein-5-isothiocyanate, FITC) and lanthanide ion (Nd3+). These changes were approximated by simple Henderson Hasselbach equation with the apparent pK 7.0 +/- 0.1 which is similar that of a histidyl residue [3]. In this work it was used the double chemical modification of SR-ATPase to research the role of histidyl residues in this conformational transition. Diethyl pyrocarbonate was used to modify the histidyl residues of the SR-ATPase. The influence of histidyl modification on the functional parameters (the rates of ATP and p-nitrophenyl phosphate hydrolysis, the Ca transport and the level of Ca2+ accumulation) was monitored by the fluorescent probes (Quin-2, chlortetracycline) using fluorescent, spectrophotometric and pH-metric measurements. In the result of these experiments it was found the appropriate conditions to carry out the second modification. The DEPC-SR-ATPase was labeled by FITC. The pH-dependent conformational changes in the active site of FITC-DEPC SR-ATPase were studied by the method of the fluorescence energy transfer between FITC and Nd3+ in the region of pH 6-8. The histidyl modification of FITC-DEPC-SR ATPase resulted in the significant shift of the curve of fluorescence energy transfer efficiency (the apparent pK > 7.5). These results suggest that the conformational transition in the active site of SR-ATPase was controlled by the histidyl residues. PMID- 8714463 TI - [Phallotoxins as modifiers in the study of muscles]. AB - This review concerns to modification of muscle contractile system functioning by phallotoxins (phalloidin and its derivatives). Binding to actin in thin filament, phallotoxins can change active tension, the spectrum of mechanical relaxation times, ATPase activity, Ca2+ sensitivity. Under some conditions these changes are very pronounced (tens of percents). As a whole phallotoxins action has the following properties: 1) muscle specificity. Qualitatively different changes have been caused by phalloidin in skinned fibers of cardiac, skeletal and smooth muscle. These differences seem to reflect, at least partially, the differences in the muscle specific proteins of the thin filament: 2) dependence on physiological state of the contractile system (thin filament activation, cross-bridge cycle kinetics); 3) specificity in action of different phalloidin derivatives. Due to such properties phallotoxins suppose to be a probe some essential steps in contractile proteins interaction and a potentially power tool for study of mechanism of muscle contraction. PMID- 8714464 TI - [Effect of verapamil and anesthetics on the level of sodium and potassium ions in frog muscles. Role of hydrophobic interactions]. AB - Verapamil, general and local anesthetics influences on ionic contents of frog sartorius and cardiac muscles in respect with these drugs hydrophobicity were studied to clarify the role of verapamil hydrophobic interactions in its physiological activity. It was found that concentration thresholds of the agents toxicities were linearly linked with their hydrophobicity (in logarithmic scale). This relationship provides support for our conclusion that verapamil, general and some local anesthetics are the members of a single drug family. It is a reason to believe that a hydrophobic mechanism of general anesthetic action on cellular structure is the same as a mechanism of verapamil activity in muscles. It is possible that verapamil is accepted as medical drug exactly due to optimum combination of high verapamil hydrophobicity with its structural complementarity to receptor site responsible for use-dependent channel blocking. PMID- 8714465 TI - [Changes in structural organization of reorganized myosin filaments from skeletal muscles of the winter-hibernating suslik Citellus undulatus during awakening]. AB - Electron microscopic observations of myosins, isolated from skeletal muscles of ground squirrels at the different states (hybernation-Mhyb; winter activity- M(act); the beginning of arousal-M(begin); the end of arousal-M(end)) have revealed the differences in the structure of their filaments. The filaments reconstituted from Mhyb, M(act) and M(end) (rectal temperature 27 degrees C) exhibit the ordered structure with the myosin heads arranged regularly (period of 14.3 nm) on the filament surface. The filaments of M(begin) (rectal temperature 12 degrees C) have irregular structure with random arrangement of the myosin head clusters alternating with regions of different length devoid of heads. To elucidate the molecular bases of the structural differences observed by us the solubility of the above myosins, the paracrystal structure of corresponding light meromyosins as well as the composition and state of myosin light chains have been studied. M(begin) appeared to have the greatest solubility in comparison with the other ones. This points to the possible changes in myosin heavy chains during hybernation and arousal although no differences have been revealed in paracrystals of light meromyosins. The amount of light chains (LC3) decreased up to 40% in Mhyb in comparison with Mact and then increased up to 60-70% in Mbegin within 1-1.5 hours of arousal. It is supposed that the rapid changes in isoforms of heavy and light chains in Mbegin lead to disturbance of regular mode of filament assembly in vitro, and can result in the lowering of muscle contraction efficiency and in the increasing of the heat release. The later can contribute to thermogenesis of the animal during arousal. PMID- 8714467 TI - [Behavior of the mitotic apparatus after ultraviolet microirradiation of the spindle pole]. AB - UV-microirradiation of the centrosome (spindle pole) alters behaviour of mitotic cell. Cell reaction strongly depends on the stage of mitosis when irradiation has been made. Irradiation of one pole at metaphase blocked cell in c-mitosis for several hours. The same or even more strong irradiation of the pole at anaphase slightly postponed cytokinesis, but didn't prevent chromosome separation and normal exit of cell into interphase. Irradiation at metaphase resulted in immediate shift of chromosomes towards the nonirradiated pole. Afterwards spindle disassembled. Irradiation at anaphase slowed down chromosome motion towards irradiated pole without prominent effect on the opposite halfspindle. Electron microscopy demonstrated that immediately after irradiation centrosome lost association with kinetochore microtubules. At metaphase irradiated pole moved towards chromosomes, while at anaphase--away from them. Same irradiation of other besides spindle poles regions had no effect on continuation of mitosis. PMID- 8714466 TI - [Patterns of adaptive mobilization of proteins from skeletal muscles during development of compensatory hypertrophy of internal organs]. AB - These studies were dealing with muscle protein depletion in rats under the heart hypertrophy induced by aorta coarctation or under liver regeneration after partial hepatectomy. The last experimental model was also used in order to establish how far such protein mobilization from muscles is realized during their simultaneous adaptation to local overload or disuse induced by a surgical method. 3 and 7 days after induction of hypertrophy of organs investigated, the quantities of sarcoplasmic, myofibrillar and stroma proteins were measured in last-twitch (m. extensor digitorum longus, m. biceps brachii) and slow-twitch (m. soleus) muscles. The contributions of protein biosynthesis and/or protein degradation change into muscle protein loss were evaluated by specially elaborated radiotracer method. The data obtained demonstrate that the pattern of muscle protein mobilization depends on nature of organ and rate of its hypertrophy. The process occurs differently in muscle of different type and in various muscle structures. It appears to be potentiated in overloaded muscles and to a lesser extent in unused ones. The loss of muscle proteins is certainly associated with inhibition of their biosynthesis. Increased degradation (or release) of preformed (non-labelled) protein molecules may also contribute to this loss whereas degradation of newly-formed (labelled) molecules does not change appreciably, possibly because their distribution in muscle structures is non-uniform. PMID- 8714468 TI - [Biomechanics of actomyosin-dependent mobility of keratinocytes]. AB - Migration of epidermal skin cells, keratinocytes, plays an important role in processes as wound healing or melanoma formation. Thus, characterization of their spontaneous motility in vitro could give indications about abnormalities of the responsible motor system, namely the cortical actomyosin layer and its interaction with the plasma membrane. Videomicroscopical observations and image data evaluations of keratinocytes spread on glass have revealed typical patterns of peripheral shape changes as rhythmical protrusions and retractions of lamellipodia, periodic waves of ruffles moving towards the cell body and, finally, pulsating or rotating waves of cell protrusions. We present two models of the behavior of actomyosin network taken as a highly viscous reactive and contractive polymer fluid. One describes the dynamics of lamellipodial protrusions transversal to the cell periphery, the other considers a simplified circular model along the whole cell circumference. Simulations of these partial differential equation models reproduce the observed spatio-temporal patterns. PMID- 8714469 TI - [Organization of mobile structures in the blood stream--basis for the function of perfluorocarbon "artificial blood"]. AB - Research into the synthetic blood substitute of the Perftoran emulsion was shown that particle dynamical chains were formed by the turbulization of a particle motility in a blood flow. This chain display an activity of the oxygen carrier channel into the tissue of vessels from the erythrocyte which remained after a bloodletting. Moreover the review presented other results of safety and efficacy of perfluorocarbon blood substitutes and theirs prognosis of future progress. PMID- 8714470 TI - [Splitting of the back edge of a propagating excitation pulse]. AB - A new type of unstability of action potential propagating in spatially inhomogeneous electrically excitable media is announced. Numerical experiments with the FitzHugh-Nagumo model show that if the medium contains a region where membrane slow, outward currents are partially depressed then the back front of action potential passing through such a region can split forming a "burning drop" solitary standing wave of depolarisation with finite "life duration". PMID- 8714471 TI - [Effect of thiol reagents on the structure of mucus glycoproteins (mucins)]. AB - The structure of both native mucins and subunits obtained by thiol reduction was studied by small-angle X-ray diffraction. X-ray pattern from the subunits significantly differs from that of the native mucins in the number, spacings and width of the reflections observed. Decrease in the number of the reflections and increase of their width pointed out that isolated subunits are less regular structure than the whole mucin molecule. This may be due to the two reasons: 1) conformation of the subunits in native molecule differs from that of in isolate state; 2) native mucin molecule is not a simple polymer, formed by the subunits. PMID- 8714472 TI - [Modification of the activity of murine peritoneal neutrophils upon exposure to millimeter waves at close and far distances from the emitter]. AB - The comparison of horn, dielectric and channel antennae on their matching with various types of loads, including a biological object, is carried out. The channel antenna in contrast to dielectric and horn ones provides the uniform spatial distribution of specific absorbed rating in the frequency range used and wide-band matching with the object both in near field and far field zones of the radiator. It is shown, that low-intensity electromagnetic radiation of extremely high frequency in near field zone of the channel radiator modifies the activity of mouse peritoneal neutrophils on a quasi-resonance manner. The interaction of electromagnetic radiation with the biological object has been revealed in the narrow-band frequencies of 41.8-42.05 GHz and consists in inhibition of luminol dependent chemiluminescence of neutrophils activated by opsonized zymosan. It is not found any frequency dependence of the electromagnetic radiation effects in the far field zone of the radiator. The results obtained suggest, that the quasi resonance dependence of the biological effect on the frequency of the electromagnetic radiation in the near field zone is conditioned by structure and nature of the electromagnetic radiation in this zone. PMID- 8714473 TI - [Resolution of fluorescence spectra by degree of quenching accessibility]. AB - The novel algorithm QUENCH is described which resolves fluorescence spectra onto two components differing in the accessibility (the Stern-Volmer constants) for small quenchers. In contrast to the known algorithms, the QUENCH first estimates Stern-Volmer constants using the total arrow of spectra measured at different quencher concentrations and, then, calculates contributions of each component into the emission intensities at every wavelength value. The component spectra, resolved with QUENCH from the tryptophan fluorescence of ribosomal S7 protein, were very similar to the log-normal components revealed using the SIMS program [16]. The QUENCH algorithm may be of use in the component analysis of fluorescence spectra of any system containing fluorophores with differing accessibilities for quenchers. PMID- 8714474 TI - [Bioeffects of weak static and alternating magnetic fields]. AB - The model for the interaction of weak magnetic fields with biosystems, which has been suggested earlier by the author for the case of impulse-like excitation of oscillators (i.e. of ions bound in the Ca(2+)-dependent enzymes or in the protein enzyme complexes) is extended for the more realistic case of continuous excitation of oscillators. The expressions for the polarization degree of oscillator's vibrations in combined, static and alternating magnetic fields are derived. It is postulated that the value of bioeffect induced by magnetic field in a biosystem is proportional to the polarization degree of the ion's vibration. The available experimental data are shown to be in a remarkably good fit with theoretical predictions. PMID- 8714476 TI - [Statistics of periodic regularities in sequences of human introns]. AB - A formula for the calculation of average power of a Fourier harmonic of a nucleotide sequence with given composition by means of the matrix Fourier analysis method is suggested. A search for the periodic regularities with the periods from two to ten nucleotides was carried out on a sample set of human exons and introns. It is shown that all the periodicities researched, including the three nucleotides period, are random in introns, whereas in exons the 3 periodic regularity is much stronger. The periods that are not multiples of 3, however, are also random in exons. The two nucleotides periodicity, which is, according to the literature, over-represented in introns, appears to be also random in the case under study. The results are important for the development of the algorithms for the coding regions search in DNA, and for the understanding of the global statistic regularities in human genome. PMID- 8714475 TI - [Ecology and problems of the search for life on Mars]. AB - Global gases-exchange metabolic cycles, which on the Earth connect autotrophic and heterotrophic organisms in indivisible biosphere of Earth, are absent on Mars as a result of rarely atmosphere. That is why Life on Mars way exist only in form of autotrophic-heterotrophic organisms-symbionts with closed metabolic cycles. Dust storms on Mars may produce continuous settling of Life in a new places which is favourable for Life. The methods of Life-searching on Mars mast take in account that fundamental difference of Life formation on Mars from so on Earth. PMID- 8714477 TI - [A specific complex between a polypeptide in the left helical conformation of polyproline type II and the B-DNA minor groove]. AB - The model of complex formation between polypeptide adopted lefthanded conformation of polyproline II type and minor groove of B-DNA presented. The specific recognition is attained due to possibility of hydrogen bond between Gly NH-group and Thymine O2-group and in addition between Pro CO-group and Guanine NH2-group and between Gly CO-group and Guanine NH2-group. Stereochemical satisfactory of the complex is observed. PMID- 8714478 TI - [Stroke in young people]. AB - We retrospectively studied 75 consecutive patients with acute stroke. They made up 3.7% of all acute cerebrovascular pathologies. Average age of patients was 37.45 (SD 7.37) and the number of men exceeded women at a ratio of 1.8:1 73% had ischaemic pathology. Those risk factors most often found were tobacco and alcohol consumption and migraine. The most frequent causes were arteriosclerosis and cardioembolism. Etiological diagnosis was not reached in a third of cases. Most patients were treated with antiagregants with aspirin being the most used at a dose of about 200 mg per day. Initial death rate was 4% and more than 60% of patients were asymptomatic or with minimal deficiencies upon discharge. PMID- 8714479 TI - [Cerebrovascular pathology: inpatient study in a community hospital. A prospective study]. AB - We present the preliminary results of six months cerebrovascular pathology follow up carried out at the Sant Boi Local Hospital as from when a Neurological Unit was set up within the Internal Medecine Service. For this purpose we drew up a prospective study based on questionnaires. A total of 69 neurological patients (2.45% of all those hospitalized) were studied, 62.3% of whom suffered acute stroke. Diagnostic and therapeutic activities were analyzed with regards to the afore mentioned pathology, as well as various data of an epidermiological nature. Lastly we present the experimental multidisciplinary approach to acute stroke as performed during hospitalization by the Internal Medecine Service, Convalescence Unit and an Interdepartmental Socio-Sanitary Functional Unit. PMID- 8714480 TI - [Intraparenchymatous hemorrhagic pathology in patients aged over seventy: a comparative study with younger subjects]. AB - The aim of our study was to compare a group of over 70 year-old patients suffering from spontaneous intracerebral haemorrhage with a similarly affected group under 70 years of age. With this in mind we carried out a prospective study of all patients admitted to the Neurology Service at the Virgen de la Arrixaca Hospital between March and December 1993. They totalled some 67 patients, 33 being over 70 years of age and 34 being younger than 70. Although our data clearly show that age is a factor indicative of bad prognosis, we found no other differences between these two groups which might justify these data with the exception of a greater prevalence of consciousness abnormalities at the moment of onset in those patients aged over 70. This fact, ascertained using the Canadian scale, may, we feel, be useful in dealing with patients presenting clinical pictures compatible with brain haemorrhage who attend hospitals not having computerized tomography. PMID- 8714481 TI - [Spontaneous dissection of the vertebral artery]. AB - Spontaneous dissection of the vertebral artery (VA) is an infrequent cause of vertebral-basilar ischaemia in children and young adults, being responsible for 4% of cases of ischaemic infarct in this age group. The distinction between spontaneous dissection and traumatic dissection helps to clarify its vascular tendency, not brought on by laceration of the vessel wall secondary to traumatism. It appears clinically with neck pain and/or headache, followed by a clinical picture of ischaemia in the vertebral-basilar area. Diagnosis is based on clinical suspicion and identification of the angiographic signs of dissection prognosis is favourable with good recuperation in 88% of cases and low recurrence risk. We present a series of seven patients with ischaemic stroke of the brainstem brought about by spontaneous VA dissection. We studied the patients using computerized tomography (CT) scan, magnetic resonance (MR) and brain angiography. The group comprised six men and one woman aged from 9 to 44 years. In one case localization was intracranial, in five there was earlier arterial pathology (hypoplasia or dysplasia) and in the remaining case dissection was bilateral. After a long-term follow-up of between one and seven years, not one of our patients showed any recurrence of ischaemic signs. We would suggest that spontaneous VA dissection should be considered in differential diagnosis in clinical pictures of vertebral-basilar ischaemia in children and young adults since most probably its frequency is greater than that currently supposed. The discussed data would support underlying arterial pathology as a tendency factor. PMID- 8714482 TI - [Eight cases of extracranial vertebral artery dissection]. AB - We present eight cases of extracranial vertebral artery dissection. One of these had traumatic antecedents at the neck level while undergoing massage treatment. Of the rest, in four cases there was only a history of commonplace traumatism at neck level, consisting of twisting or stretching. Most presented pain at this level both before and during symptoms. Five had symptoms compatible with lateral bulbar infarct, two with cerebral infarct and one at the protuberance level. Angiography showed irregular stenosis of the spine on the affected side in five cases, occlusion in three cases. Nuclear magnetic resonance (NMR) was performed on five, with findings compatible with dissection. Six received anticoagulant treatment and two received platelet antiagregants with good recovery except in one patient who died twelve months later without any indication of the existence of dissection. We also carried out a review of the literature with special emphasis on the etiology of spontaneous cases, on clinical and neuroimaging findings and on treatment. PMID- 8714483 TI - [Stroke and hematological disorders in young people]. AB - In this work we analyse the information about one group of young patients with cerebrovascular disease, studying the association between haematological pathology and cerebrovascular disease. The objective is to establish a comparison between our results and those previously reported in the medical literature. PMID- 8714484 TI - [Clinical pathogenic heterogeneity and subcortical infarcts prognosis]. AB - We present a descriptive study of 55 consecutive patients with subcortical infarct seen during the year 1993. We divided our sample into two groups: the first included a total of 49 patients with lacunar infarct (ischaemic lesion equal to or less than 20 mm in diameter or lacunar syndrome with no objective lesion). The second group consisted of six patients with subcortical infarct greater than 20 mm at its largest diameter. We touch upon vascular risk factors, probable etiology, clinical history and prognosis, trying to determine as to whether differences existed between the two groups or not. The sole case showing statistically significant difference was the clinical presentation syndrome although results are hardly conclusive given the small size of our sample groups. PMID- 8714485 TI - [Cerebral hemorrhage and migraine]. AB - The connection between migraine and brain haemorrhage is controversial. We present the case of eight nonhypertense patients all aged under 57 with migraine antecedents who suffered brain haemorrhage during an attack. All underwent analytical study, chest X-ray, electrocardiography, computerized tomography scan and brain panangiography. The study was completed in six cases with an immunological analysis and in a further five with brain magnetic resonance. Seven patients habitually took vasoactive drugs to relieve migraine. The results do not show any other cause of brain haemorrhage. It is possible haemorrhage may be related to vascular lesion brought about by ischaemia secondary to vasospasms. PMID- 8714486 TI - [The prognostic value and evolution of blood pressure during an acute phase of stroke]. AB - We studied the prognostic influence and evolution of blood pressure during the acute phase of stroke in 89 patients (50 men and 39 women) with an average age of 69.4 +/- 10.8 years. Seventy-two were diagnosed as having ischaemic infarct and 17 as having spontaneous intracerebral haemorrhage. Blood pressure was taken every four hours for twelve days. Clinical situation was evaluated using the Rankin scale. Systolic and diastolic blood pressure progressively decreased without needing any medication in the first two weeks of evolution. The decrease was greatest in hypertense patients and in those with left ventricle hypertrophy. We found the initial figures for systolic and diastolic blood pressure significantly higher in those patients with brain infarct who had not died and in those in a better functional position the second week of evolution. Blood pressure did not influence the prognosis of intracerebral haemorrhage patients. PMID- 8714487 TI - [The prognostic value of analytical hemorheological factors in stroke]. AB - Plasmatic hyperviscosity is a known vascular risk factor which depends, among other parameters, upon total proteins, haematocrit value and fibrinogen. The aim of this study was to determine the relationship between these analytical haemorrological parameters and mortality and acute stroke sickness rate. We studied 216 patients with brain infarct and spontaneous intracerebral haemorrhage out of a sample of 256 acute stroke patients. We evaluated neurological affectation using the Canadian scale upon admission, on the seventh day and after three months. We determined plasmatic haemorrological factors (haematocrit, fibrinogen and total proteins) upon admission and again after one week and then at three months High fibrinogen levels in plasma were related to brain infarct mortality regardless of the cause of death. PMID- 8714488 TI - [Predictive mortality factors and the development of major complications in non traumatic subarachnoid hemorrhage]. AB - Our aim was to analyze death prognostic factors in the first six months or the development of major neurological complications (delayed brain ischaemia and rebleeding) or general complications (cardiorespiratory, hydroelectrolytic or infectious) in non-traumatic subarachnoid haemorrhage. We carried out an observational study of a historical cohort of 111 patients admitted consecutively to our Centre between 1986 and 1992 and who were diagnosed as having subarachnoid haemorrhage based upon clinical data and confirmed in all cases by brain computerized tomography (CT) scan and/or lumbar puncture. Clinical variables upon admission were collected prospectively and radiological data were obtained retrospectively. We performed a logistic regression analysis which showed as significant predictive variables for death in the first six months a score on the Hunt-Hess scale greater than 2, a score on the Glassow scale of less than 9, female, aged over 50 and rebleeding. As predictive factors for delayed brain ischaemia we identified the female sex and the existence of earlier general complications. In the development of rebleeding a significant predictive factor was high blood pressure antecedents although this may have been controlled upon admission. For the appearance of general complications, significant predictive factors were more than 2 on the Hunt-Hess scale and hypertension antecedents. The use of these predictive parameters may help evaluate a prognosis in subarachnoid haemorrhage patients and also help make decisions in the management of this pathology. PMID- 8714489 TI - [3 year survival in patients hospitalized for acute cerebrovascular disorders]. AB - For three years we studied the mortality and functional situation of all patients admitted in 1991 to the Neurology Service suffering from acute stroke with the exception of subarachnoid haemorrhage cases. We analyzed the cause of death whether directly related to the initial illness or not. Out of 134 patients admitted for acute stroke, 48 (41.02% of the 117 patients examined after excluding 17 whom we did not obtain complete information from) had died after three years. The main causes of death were directly related to acute stroke (37.5%) and pneumonia (37.5%). Death occurred mainly in the first month (79.16% of deaths). Predictive variables for mortality directly related to acute stroke during the first month include severe weakness, brain haemorrhage, dysphasia and earlier incidence of acute stroke. Variables related to higher mortality rate due to other causes in the first month were dysphasia, age and angina antecedents, whereas earlier incidence of acute stroke was associated with a lesser mortality rate for these causes, as distinct from acute stroke itself. Greater levels of weakness and sphincteral incontinence are the best predictive signs of dependency functional situation at the end of the first month and, along with diabetes, after one and three years. PMID- 8714490 TI - [The advantages of associating ischemia/reperfusion injury inhibitors with intravenous thrombolysis when treating focal brain ischemia: a description of experiments using rats]. AB - Restoration of blood flow is necessary when treating brain ischemia. However, except in very early cases, this is insufficient to prevent the cascade of mediators of cell damage unleashed by ischemia which, on the other hand, is boosted by the deleterious effects of reperfusion. We therefore consider adequate treatment for brain ischemia ought to associate, early on, reperfusion with pharmacological inhibition of those intermediaries in damage caused by ischemia/reperfusion (cytoprotection), basically an excess of cytosolic calcium and of free radicals. In this way, spreading of the infarct may be avoided more effectively than just with reperfusion alone. Our aim was to demonstrate this hypothesis using an experimental model. In order to do so, we shall embolize the right carotid artery territory of 50 Long Evans rats with autologous blood thrombi and check the location of the embolus using arteriography. Forty rats will receive thrombolytic treatment intravenously (rTPA at a dose of 20 mg/kg) two hours after embolization, while 10 control rats will receive similar treatment with saline serum. The thrombolysis treated animal group will be divided into four subgroups (A, B, C, D, ni = 10). Group A will be considered as the thrombolysis control group; group B will additionally receive calcium antagonists (dihydropyridines); group C will undergo antioxidant treatment (21 aminosteroids) and group D both calcium antagonists and antioxidants in association with thrombolysis. The size of infarct produced in each group, estimated following Cavalieri's principle, will be compared using nonparametric statistical tests (Mann-Whitney test and Willcoxon test). We present the preliminary results so obtained in a group of 14 control rats studied with the aim of assessing the usefulness of the proposed model. PMID- 8714491 TI - [Antiaggregant treatment for cerebral ischemia: ticlopidine versus aspirin]. AB - INTRODUCTION: There at present exists marked controversy as to the possible greater efficacy of ticlopidine as opposed to aspirin in brain ischaemia secondary prophylaxis. In our study we compare the efficacy and safety of antiaggregant treatment with ticlopidine as opposed to aspirin in a group of 310 patients admitted to the 'Hospital Mutua de Terrassa' between the years 1990 and 1994. RESULTS: In the group of patients treated with ticlopidine we found a larger number of new cerebrovascular incidents (p = 0.02) and peripheral vascular incidents (p = 0.01). New cerebrovascular incidents were more frequent in males (p = 0.03), in those patients with substantiated infarct (p = 0.02) and in patients with ischaemia in the carotidal region (p = 0.04). On the other hand, the group of patients treated with ticlopidine presented more secondary effects than the group treated with aspirin at the digestive (p < 0.001) and haematologic (p < 0.001) levels. The most frequent digestive secondary effects were diarrhoea (p < 0.001) and hepatopathy (p = 0.02); abnormalities in the leucocyte count were more frequent in patients treated with ticlopidine (p < 0.001), neutropenia being found in 0.8% of cases. CONCLUSION: In the patients we studied ticlopidine was less efficacious than aspirin in the secondary prophylaxis of new vascular incidents (both cerebrovascular and peripheral vascular) and also presented a greater incidence of secondary (digestive and haematological) effects. PMID- 8714492 TI - [A retrospective study of potential benefits of thrombolytic drugs in a stroke]. AB - We carried out a retrospective analysis into the possible uses of thrombotic drugs on 190 consecutive patients with ischaemic stroke who had been admitted to the Neurological Service of the Barcelona 'Hospital de la Santa Creu i Sant Pau' in accordance with exclusion criteria commonly accepted for the use of this type of medication. We thus analyzed the initial symptoms, type of instauration, etiological diagnosis and topographical diagnosis. In those patients who could be possible candidates for such treatment with these drugs we assessed their disability variations (on the Rankin scale) at the end of two weeks using currently available therapeutic means. 70% of patients would subsequently be excluded from our study. Among reasons for exclusion were especially minor neurological deficiencies, age and latency time. As for patients included, most predominant factors were immediate instauration, cardioembolic etiological mechanism and severe onset symptomatology (motor disorder, awareness and language abnormalities). We observed a substantial spontaneous improvement in 7% of patients. This fact is especially clear in cardioemboligenic etiology patients. Bearing the spontaneous improvement cases in mind, we concluded that some 23% of patients with ischaemic cerebrovascular pathology could benefit from thrombotic drugs in our milieu. PMID- 8714495 TI - [Who should coordinate a society for sleep disorders?]. PMID- 8714493 TI - [The effect of angioplasty on cerebral vasomotor reactivity in patients with important stenosis of the internal carotid artery]. AB - INTRODUCTION: We present the preliminary results of the effects on percutaneous translumenal angioplasty (PTA) cerebral haemodynamics in thirty patients with important stenosis of the internal carotidal artery. MATERIALS AND METHODS: Using transcranial doppler (TCD) with acetazolamide (ACZ) as a vasodilatory stimulus, we assessed vasomotor reactivity (VR and pulsatility index (Pl) both before and three months after angioplasty. We also carried out single photon emission computerized tomography (SPECT) and cartography and frequency measurement by electroencephalogram (EEG) again both before and after PTA. RESULTS: Of the first twelve patients who completed the study, VR was maintained in nine and PI increased after PTA in seven. In three patients VR was exhausted and in two recovered after PTA. CONCLUSIONS: PTA improved cerebral haemodynamics. Similar more exhaustive studies are needed if revascularization techniques are to be useful in treating patients with this level of stenosis but who are asymptomatic as to the risk of haemodynamic cerebral vasomotor angioplasty. PMID- 8714494 TI - [Who should coordinate a society for sleep disorders?]. PMID- 8714496 TI - [Who should coordinate a society for sleep disorders?]. PMID- 8714497 TI - 'STAR' war against coronary thrombosis. PMID- 8714498 TI - Unexplained syncope: diagnosis, mechanisms and treatment. PMID- 8714499 TI - Proximal flow convergence method in the assessment of mitral stenosis. AB - Mitral valve area (MVA) was calculated using the proximal flow convergence method in 60 patients with severe mitral stenosis. Using the apical 4 chamber view, colour Doppler was used to measure the peak forward flow rate. From this, the mitral valve area was calculated according to the formula: mitral valve area = peak forward flow rate/peak mitral velocity. This method compared well with the established 2D planimetry derived valve area (r = 0.89) and Doppler pressure half time method (r = 0.88). Hence, this can be used as an alternative method to estimate the valve area in patients with mitral stenosis. PMID- 8714500 TI - Risk factors for coronary heart disease in noninsulin dependent diabetes mellitus (NIDDM). AB - Plasma levels of fasting and post-prandial blood sugar, serum levels of total cholesterol (TC), triglycerides (TG), HDL cholesterol (HDLc), LDL cholesterol (LDLc) and free fatty acids (FFA) were estimated in 213 diabetic patients (NIDDM) with coronary heart disease (CHD-group 4), 252 CHD patients (non-diabetic CHD group 3), 164 non-insulin dependent diabetics (NIDDM-group 2) and 173 healthy subjects (controls-group 1) who did not have any clinical evidence of CHD, diabetes mellitus or any family history of the above diseases. Data was analysed by ANOVA along with the Duncan procedure and multiple logistic regression. Lipid profile of diabetic CHD patients was characterised by significantly higher concentration of TC, TG, LDLc, FFA, LDLc/HDLc ratio and lower concentration of HDLc. However, in a multivariate logistic regression analysis using 14 known risk factors, diastolic blood pressure (BP), body mass index (BMI), alcohol consumption and higher FFA levels seemed to be predictors of CHD in diabetics, overriding the influence of lipoprotein abnormalities. The same was true for nondiabetic patients also in whom BMI, FFA and alcohol consumption were found to be significant predictors of CHD. Thus, even though lipid abnormalities are more prominent in diabetics, the coexistence of obesity and hypertension seem to be important factors in diabetics for the development of CHD. PMID- 8714501 TI - Plasma insulin levels in coronary artery disease. AB - Seventy two consecutive patients without a history of diabetes and normal fasting plasma glucose were included in this study of insulin levels. Standard oral glucose tolerance test with 75 gm glucose and fasting and two hour insulin levels were estimated in all patients. Coronary artery disease (CAD) was confirmed or excluded by selective coronary arteriography. In 20 patients, CAD was diagnosed by electrocardiographic (ECG) and clinical evidence of earlier myocardial infarction. Mean fasting plasma insulin was 31.40 +/- 22.2 IU/dl in the CAD positive and 32.3 +/- 13.6 IU/dl in the CAD negative group. The mean two hour plasma insulin was 274.6 +/- 301.1 IU/dl in the CAD positive and 104.8 +/- 74.9 IU/dl in the CAD negative group (p < 0.04). Two hour plasma insulin levels were significantly higher in patients with atherosclerotic coronary artery disease. It is concluded that the estimation of a two hour plasma insulin level after 75 gm of glucose load, could help differentiate CAD from normals. PMID- 8714502 TI - PTCA, thrombolysis or atherectomy: rational treatment of recently occluded saphenous vein grafts. AB - Recently occluded saphenous vein grafts (SVG) contain abundant thrombus. Distal embolization and myocardial infarction often occur when recanalization of such SVG is attempted. In 80 patients with occluded SVG, we employed transcatheter devices to lyse, compress or extract thrombus. Primary treatment for these SVG was performed in the following manner; PTCA 29, intragraft urokinase 12, TEC atherectomy 39. Following urokinase or atherectomy, adjunctive PTCA was performed to diminish the residual stenosis. All patients had class III or IV angina. Clinically, SVG occlusions were 3 days to 3 months old. TIMI flow was grade 0, and occlusion length was greater than 6 cm for all SVG. Each strategy resulted in a similar procedure success rate. However, when used as a primary treatment, TEC may be associated with lower rates of distal embolization and myocardial infarction. PMID- 8714503 TI - Comparison of anti-ischemic effects of isosorbide-5-mononitrate, metoprolol and diltiazem: a study based on serial treadmill exercise test. AB - A prospective double blind cross over study compared the anti-ischemic effect of 20 mg of isosorbide-5-mononitrate (IS-5-MN), 60 mg of diltiazem and 100 mg of metoprolol by performing serial treadmill tests (TMTs) following oral administration of single dose of each drug in 28 patients of ischemic heart disease with positive exercise TMT. The anti-ischemic effect of IS-5-MN, diltiazem and metoprolol was evaluated by assessing the time to onset of ischemia (400.8 +/- 41.1 vs 394.1 +/- 36.0 vs 412.7 +/- 36.6 secs respectively, all p > or = 0.05), duration of ischemia (390.9 +/- 58.5 vs 447.6 +/- 65.9 vs 419.4 +/- 58.7 secs respectively, all p > or = 0.05), duration of exercise 464.0 +/- 32.9 vs 476.8 +/- 32.5 vs 502.6 +/- 31.3 secs respectively, all p > or = 0.05) and severity of ischemia by scoring system (10.0 +/- 1.5 vs 10.7 +/- 1.4 vs 9.8 +/- 1.5 respectively, all p > or = 0.05). Thus, the present study suggests that there is no significant difference in the anti-ischemic effect of IS-5-MN, diltiazem and metoprolol and also suggests that the anti-ischemic effect of metoprolol is primarily by decreasing the heart rate while IS-5-MN and diltiazem exert anti ischemic effect probably by decreasing oxygen demand and improving coronary blood flow. PMID- 8714504 TI - Radionuclide (MUGA) studies of left ventricular functional abnormalities in asymptomatic patients with sickle cell anemia. AB - Radionuclide ventriculography was performed on 10 normal subjects and 39 patients with sickle cell anemia (10 homozygous and 29 heterozygous sicklers) at rest and after exercise. Their left ventricular (LV) function was assessed in both these situations. The results were then compared within the subgroups. The reduction in ejection fraction (EF) response (47.5 +/- 7 at rest and 46.4 +/- 8 at exercise in homozygous patients, and 52.4 +/- 8 at rest and 54.3 +/- 8 at exercise in heterozygous patients) was significant in both the homozygous and the heterozygous groups but more so in the former group. The diastolic filling was also significantly impaired in the homozygous group (PER 2.64 +/- 0.74, PFR 2.13 +/- 0.42 and PFR/HR 0.014 +/- 0.001). The study statistically demonstrates, that LV filling patterns are altered in the sickle cell patients, even in the absence of clinical symptoms relating to LV dysfunction. This fact may prove to be a marker of sickle cell heart disease. Frequent and significant sickling is probably the cause of more pronounced LV functional abnormalities in homozygous sicklers. PMID- 8714505 TI - Primary stenting for unprotected isolated left main ostial stenosis. PMID- 8714506 TI - Percutaneous balloon aortic valvuloplasty with anterograde crossing of aortic valve. PMID- 8714507 TI - Subaortic left ventricular aneurysm presenting with complete heart block. PMID- 8714508 TI - Left ventricular myxoma with adhesions to mitral valve leaflets--case report. PMID- 8714509 TI - Mitral valve aneurysm without infective endocarditis. PMID- 8714510 TI - Radiofrequency catheter ablation of intra-atrial reentrant tachycardia. PMID- 8714511 TI - Prenatal diagnosis of cardiac rhabdomyoma. PMID- 8714512 TI - Angiotensin converting enzyme inhibitors: present status and future perspectives. PMID- 8714513 TI - National PTCA registry of India--1994. PMID- 8714514 TI - Trials for treatment of myocardial infarction ending in tribulation. PMID- 8714515 TI - Distinct positive and negative regulatory elements control neuronal and hepatic transcription of the human transferrin gene. AB - Transferrin (Tf), the iron-transport protein, plays an essential role in the central nervous system development, plasticity, and aging. As a first step toward elucidating the role of each transcription factor involved in the regulation of Tf gene expression, we have recently shown that similar promoter elements direct cell-type specific transcription in oligodendrocytes, epithelial choroid plexus cells, and in the neuronal cell line B103. Here we have analyzed the regulatory elements that control the level of expression of the Tf gene in neuronal cells. Transient expression experiments in B103 cells revealed that the -164/+1 promoter region is stimulated by a position-dependent -1140/-1000 upstream region. DNase I footprinting, gel retardation assays, and antibody reactivity data allowed us to characterize the nuclear factors interacting with this region. The upstream region I-binding protein (URI-BP) belongs to the steroid/retinoid receptor family, while URII-BP is a member of the nuclear factor I (NF-I) family. Interestingly, no enhancer nor silencer activity is detected in B103 cells. This contrasts with our findings in hepatoma cells, where the activity of the -125/+1 promoter can be repressed by a -1000/-819 upstream negative-acting region and stimulated by the -3600/-3300 enhancer. We demonstrate that the negative-acting region presents the characteristics of a silencer that interacts with a nuclear protein present in liver and absent in B103 cells. Similarly, B103 cells lack a nuclear protein able to bind to an essential site of the enhancer. This shows that in B103 cells, the inactivity of the silencer and the enhancer regions results from the absence of at least one essential nuclear protein. PMID- 8714516 TI - Monoclonal autoantibody SCH94.03, which promotes central nervous system remyelination, recognizes an antigen on the surface of oligodendrocytes. AB - A monoclonal antibody SCH94.03, made in syngeneic mice by injection of spinal cord homogenate, promotes central nervous system remyelination when injected into SJL/J mice chronically infected with Theiler's virus. To elucidate the mechanism of antibody-mediated remyelination, SCH94.03 was investigated by immunocytochemistry, flow cytometry, immunoelectron microscopy, Western blotting, and immuno-thin layer chromatography. All cell types investigated in vitro showed strong cytoplasmic staining with a pattern resembling a cytoskeletal protein. In contrast, among the primary cultured cells studied, only oligodendrocytes showed strong surface reactivity. Other cell types, including astrocytes, microglia, Schwann cells, myoblasts, and T and B lymphocytes, were negative. Mouse and rat oligodendrocytes which showed strong surface reactivity exhibited a well differentiated morphology, and approximately 50% expressed myelin basic protein. Since oligodendrocyte progenitors were negative for surface staining, the expression of the antigens recognized by this monoclonal antibody appears to be developmentally regulated, i.e., transiently expressed on younger, terminally differentiating oligodendrocytes. Among the cell lines studied, only two rat oligodendrocyte lineage cell lines showed surface reactivity with SCH 94.03. Western blotting of secondary isolated oligodendrocytes lysates revealed reactivity with multiple protein bands of 27, 32, 50, 100, and 106 kDa, whereas there was no reactivity to lipid antigens by immuno-thin layer chromatography. These results raise the possibility that SCH94.03 recognizes a novel oligodendrocyte-specific surface antigen, and may act directly on oligodendrocytes to promote remyelination. PMID- 8714517 TI - Inhibition of long-term potentiation development in rat hippocampal slice by alpha 2-macroglobulin, an acute-phase protein in the brain. AB - Alpha-2-macroglobulin (alpha 2M) in the rat and human brain is an acute-phase protein synthesized primarily by astrocytes, and it has been implicated in Alzheimer's disease and other neuropathological processes. The activated forms of alpha 2M, but not the native form, can suppress the neurite outgrowth of the central neurons, presumably through binding to neurotrophic factors and through direct inhibition of neurotrophic factor receptor signal transduction. Since neurotrophic factors are known to be involved in synaptic plasticity, we tested the effect of both the native and methylamine-activated (MA-alpha 2M) forms of alpha 2M on long-term potentiation (LTP) in area CA1 of adult rat hippocampal slice. Neither native alpha 2M nor MA-alpha 2M had an effect on baseline synaptic transmission. LTP induced by 200-Hz trains in the presence of 1.4 microM or 0.14 microM native alpha 2M was indistinguishable from control LTP. Although the presence of MA-alpha 2M at the same concentrations did not interfere with LTP induction, the development and maintenance of potentiation was blocked in a concentration-dependent time course. Results of this study indicate that the accumulation and activation of alpha 2M with inflammatory neuropathologies such as Alzheimer's disease can inhibit synaptic plasticity, which might partly account for the memory deficits seen in these patients. PMID- 8714518 TI - Ciliary neurotrophic factor selectively protects human oligodendrocytes from tumor necrosis factor-mediated injury. AB - Oligodendrocytes (OLs) and their myelin membranes are the apparent injury targets in the putative human autoimmune disease multiple sclerosis. The basis for this selective injury remains to be defined. OLs in vitro have been shown to be susceptible to both tumor necrosis factor (TNF) and non-TNF-dependent immune effector mechanisms. The former involves initial nuclear injury (apoptosis); the latter, when mediated by activated T cells, involves initial cell membrane injury (lysis). In the current study, we determined whether human adult CNS-derived OLs could be protected from the above immune effector mechanisms by selected neurotrophic factors (CNTF, BDNF, NGF, NT-3, and NT-4/5) or cytokines demonstrated to protect from human or experimental autoimmune demyelinating diseases (beta-interferon [IFN], IL-10, and TGF-beta). Nuclear injury was assessed in terms of DNA fragmentation using a DNA nick-end-labelling technique; cell membrane injury was assessed by lactate dehydrogenase or chromium 51 release. MTT and cell counting assays were used to assess cell viability and cell loss, respectively. Amongst the neurotrophic factors and cytokines tested, only CNTF significantly protected the OLs from TNF-mediated injury. CNTF also protected the OLs from serum deprivation-induced apoptosis. CNTF, however, did not protect the OLs from injury induced by activated CD4+ T cells. CNTF also did not protect human fetal cortical neurons from serum deprivation or TNF-induced DNA fragmentation, nor did it protect the U251 human glioma cell line from DNA fragmentation induced by a combination of TNF and reduced serum concentration in the culture media. Our results indicate that potential protective effects of neurotrophic factors or cytokines on neural cell populations can be selective both for cell type involved and mechanism of immune-mediated injury. CNTF is the protective factor selective for nuclear-directed injury of OLs. PMID- 8714519 TI - Co-localization of NG2 proteoglycan and PDGF alpha-receptor on O2A progenitor cells in the developing rat brain. AB - A detailed comparison in the developing rat central nervous system between the distribution of the NG2 proteoglycan and the alpha-receptor for platelet-derived growth factor (PDGF) shows that these two molecules are co-expressed by glial progenitor cells of the O2A lineage and can serve as reliable markers for identification of O2A cells in vivo. Our mapping experiments indicate that NG2 positive, PDGF alpha-receptor positive O2A cells are abundant throughout the developing central nervous system in both white and gray matter. The earliest cells immunoreactive for either of the two markers are found adjacent to the central canal of the embryonic day 15 (E15) spinal cord. These cells express only PDGF alpha-receptor and not NG2. By E17, process-bearing cells expressing both NG2 and PDGF alpha-receptor in a highly co-localized fashion are found throughout the central nervous system. The first postnatal week marks the peak in the number of NG2 and PDGF alpha-receptor immunoreactive cells, as well as the peak in the level of expression and the extent of co-localization of the two molecules. After the first week, the level of expression of both NG2 and PDGF alpha-receptor declines, although both molecules continue to be expressed in the adult brain. On O2A cells in the mature brain, NG2 and PDGF alpha-receptor are not as well co localized at the subcellular level as they are on O2A cells in the younger brain. The functional consequences of co-localization and subsequent dissociation of NG2 and PDGF alpha-receptor on maturing O2A progenitors are investigated in the accompanying paper (Nishiyama et al.: J Neurosci Res 43:315-330, 1996). PMID- 8714520 TI - Interaction between NG2 proteoglycan and PDGF alpha-receptor on O2A progenitor cells is required for optimal response to PDGF. AB - Previous studies on the NG2 chondroitin sulfate proteoglycan have shown that NG2 is expressed on A2B5-positive O2A progenitor cells, which are known to respond to platelet-derived growth factor (PDGF). In the accompanying paper (Nishiyama et al.; J Neurosci Res 43:299-314, 1996) we show that on O2A progenitors in the embryonic and newborn rat brain, NG2 and PDGF alpha-receptor display an extensive co-localization which becomes less pronounced as the brain matures past the first postnatal week. The present communication describes the relationship between NG2 and PDGF alpha-receptor in vitro. NG2 and PDGF alpha-receptor are highly co localized on A2B5-positive O2A cells isolated from neonatal rat cerebrum. Mimicking the situation in vivo, the level of expression of the two molecules and the extent of co-localization decline as these cells differentiate into O4 positive pre-oligodendrocytes. However, maintenance of the cells in a progenitor state by treatment with bFGF results in increased levels of both NG2 and PDGF alpha-receptor on the cell surface, suggesting that expression of the two molecules may be coordinately regulated. Furthermore, NG2 can be co immunoprecipitated from radiolabeled O2A extracts with a rabbit antibody to PDGF alpha-receptor, indicating the presence of a molecular complex that includes NG2 and the receptor. Finally, antibody-patching and subsequent down-regulation of NG2 results in reduced expression of PDGF alpha-receptor and diminishes the proliferative response of the cells to PDGF. These findings suggest that correct co-expression of the NG2 proteoglycan and PDGF alpha-receptor on the surface of O2A progenitor cells is important for the cells' ability to respond effectively to the mitogenic stimulus of PDGF. PMID- 8714521 TI - Dopamine differentiation factors increase striatal dopaminergic function in 1 methyl-4-phenyl-1,2,3,6-tetrahydropyridine-lesioned mice. AB - We have previously shown that muscle-derived differentiation factors (MDF) and human recombinant acidic fibroblast growth factor (aFGF) have beneficial behavioral and neurochemical effects on the nigrostriatal dopaminergic neurons of 6-hydroxy-dopamine (6-OHDA)-lesioned rats (Jin and Iacovitti: Neurobiol Dis 2:1 12, 1995). In the present study, we determined the effects of similar treatments on mice treated with the neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). Five days after unilateral striatal infusion of MDF or aFGF into MPTP lesioned mice, striatal tyrosine hydroxylase (TH) activity and dihydroxyphenylacetic acid (DOPAC) levels were bilaterally increased (20-35%) compared to untreated (lesion only) or control (phosphate buffered saline + bovine serum albumin) mice. These increases, however, were not accompanied by change in dopamine (DA) levels, indicating an elevation of DA synthesis (TH/DA) and turnover (DOPAC/DA). The present findings that MDF and aFGF may have neurochemical effects in vivo on the lesioned nigrostriatal dopaminergic system suggest their potential pharmacological role in the treatment of Parkinson's disease. PMID- 8714522 TI - Expression of mRNA species encoding heat shock protein 90 (hsp90) in control and hyperthermic rabbit brain. AB - Northern blot and in situ hybridization were employed to investigate regional and cell type differences in the expression of hsp90 mRNA species in control and hyperthermic rabbit brain. Riboprobes specific to hsp90 alpha and beta mRNA species were utilized in time-course Northern blot studies on cerebral hemispheres and the cerebellum. Following hyperthermia, levels of hsp90 alpha and beta mRNA were elevated in both brain regions; however, the magnitude of induction was more robust in the cerebellum than in cerebral hemispheres. The pattern of expression of hsp90 genes in rabbit brain was analyzed by in situ hybridization. These studies revealed that hsp90 genes are preferentially expressed in neuronal cell populations in the unstressed mammalian brain. The distribution of hsp90 alpha and beta mRNA was similar, though the signal for the latter was stronger. Following hyperthermia, changes were not detected in the pattern of hsp90 beta mRNA expression in the hippocampus. In the cerebellum, a rapid induction of hsp90 beta mRNA was apparent in the neuron-enriched granule cell layer, followed by a delayed accumulation in Purkinje neurons. Unlike hsp70, induction of hsp90 was not detected in glial cells of hyperthermic rabbit brain. The localization of hsp90 to neurons suggests that this heat shock protein plays an important role in neuronal function. PMID- 8714523 TI - Glycine increases the number of somatostatin receptors and somatostatin-mediated inhibition of the adenylate cyclase system in the rat hippocampus. AB - The glycine and somatostatin (SS) neurotransmission systems in the brain have been implicated in the function of sensory, motor, and nociceptive pathways. To investigate a possible relationship between these two components, we studied the influence of glycine on the binding of 125I-Tyr11-SS to its receptors and on SS like immunoreactivity (SSLI) levels in the rat hippocampus and frontoparietal cortex. An intracerebroventricular (i.c.v.) dose of 16 or 160 nmol of glycine induced an increase in the total number of specific SS receptors in the hippocampus but not in the frontoparietal cortex at 15 min following injection, with no changes in the affinity constant. This effect seems to be mediated by inhibitory strychnine-sensitive glycine receptors since pretreatment with the antagonist strychnine (80 micrograms/100 g body weight, intravenously) abolished this response. No significant changes in SSLI content were detected in either brain region of glycine- and strychnine plus glycine-treated rats as compared to control values. Since SS receptors are coupled via guanine nucleotide-binding G proteins to the adenylyl cyclase (AC) system, we also examined the inhibitory effects of SS and the guanine nucleotide Gpp(NH)p on AC activity in hippocampal membranes of control, glycine- and strychnine plus glycine-treated rats since the increase in SS receptors was observed only in this brain area. No significant differences were observed for the forskolin (FK)-stimulated AC enzyme activities in hippocampal membranes from all the experimental groups studied. In the hippocampus of the glycine- (160 nmol) treated group, however, basal AC activity was significantly lower, and the capacity of SS to inhibit FK-stimulated AC activity was increased as compared to the control group. Pretreatment with strychnine prevented the increase in SS-mediated inhibition of AC activity. The functional activity of the inhibitory guanine nucleotide-binding protein Gi, as determined by the inhibitory effect of the stable GTP analogue Gpp(NH)p on FK stimulated AC activity, was significantly higher in hippocampal membranes of glycine- (160 nmol) treated rats as compared to controls. This suggests that the increased inhibition of AC activity by SS in the glycine-treated group may be due to the increase in Gi activity and/or the increase in the number of SS receptors observed. Alternatively, the greater Gi activity may be responsible for the increased binding of 125I-Tyr11-SS to its receptors observed after glycine administration. Altogether, these data suggest that the hippocampal somatostatinergic system can be regulated by strychnine-sensitive glycine receptors in the rat. PMID- 8714524 TI - PC12h-R cell, a subclone of PC12 cells, shows EGF-induced neuronal differentiation and sustained signaling. AB - Unlike nerve growth factor (NGF), epidermal growth factor (EGF) does not induce neuronal differentiation but promotes proliferation of the rat pheochromocytoma PC12 cells. We found that PC12h-R, a subclone of PC12 cells, differentiated into neuron-like cells in response to EGF as well as to NGF. PC12h-R cells treated with EGF extended neurites, attenuated cell proliferation, and increased the levels of tyrosine hydroxylase protein synthesis and of acetylcholinesterase activity as those treated with NGF. The EGF-induced differentiation of PC12h-R cells was not mediated by the indirect activation of p140trkA by EGF. In addition, EGF induced the sustained tyrosine phosphorylation of the EGF receptor, mitogen-activated protein (MAP) kinases, and 46 and 52 kDa proteins, and the prolonged activation of MAP kinases in PC12h-R cells compared with the parent PC12h, which does not show EGF-induced differentiation. The response of PC12h-R cells to EGF was not simply due to an increase in the level of EGF receptor protein. These results indicated that the duration of EGF-induced signaling might determine the cellular response of PC12 cells between cell proliferation and neuronal differentiation. PMID- 8714525 TI - In vitro accumulation of glucocerebroside in neuroblastoma cells: a model for study of Gaucher disease pathobiology. AB - Gaucher disease is the most common lysosomal glycosphingolipid storage disease; decreased activity of glucosylceramide beta-glucosidase (GCase) results in the accumulation of glucocerebroside (GlcCer) in macrophage-derived cells. The most devastating types of Gaucher disease also involve neuronopathology, thought to be mediated by intracellular GlcCer accumulation in the brain. In this study, we developed an in vitro neuronal cell model for accumulation of endogenous GlcCer to enable studies on the cellular basis for the neuronopathology of this disease. A human neuroblastoma cell line (SH-SY5Y) was selected because it produced appreciable GCase. When these cells were treated with conduritol B epoxide (CBE), a competitive, irreversible inhibitor of this enzyme, GCase levels fell precipitously, while other lysosomal hydrolase levels were unaffected. Relative to untreated control cells, the CBE-treated cells accumulated higher levels of GlcCer, but not other related glycolipids, over time. Thus, this in vitro system displayed many essential biological parameters relevant for studies on cellular events responsible for the neurologic damage that occurs in some types of Gaucher disease. This model should also be useful in investigations of the normal role of sphingolipids in neuronal cell function. PMID- 8714527 TI - Central glial and neuronal populations display differential sensitivity to ceramide-dependent cell death. AB - Ceramide is a lipid second messenger implicated in the mechanism of apoptotic cell death. The effect of the cell-permeable ceramide analogue C2 has been tested on primary cortical cultures of neurons, astrocytes, and oligodendrocytes as well as on the bipotential glial precursor cell line CG-4. After 24 hr of treatment, C2 ceramide induced a dose-dependent cell death in primary oligodendrocytes and precursor cells, with a maximum effect at 10 microM. Commitment of oligodendrocytes to cell death occurred within the first 6 hr of treatment. Ultramicroscopic analysis of primary oligodendrocytes exposed to C2 ceramide for 3.5 hr revealed extensive membrane blebbing in the absence of nuclear condensation. In contrast, similar treatment of primary neuronal or astrocytic cortical cultures had no effect on cell survival. Neurons and astrocytes were resistant to 10 microM C2 ceramide. Furthermore, bipotential progenitors that were differentiated toward astrocytes also became resistant to ceramide treatment as they acquired a mature astrocytic phenotype. These experiments suggest that cell type specific factors are required for ceramide-mediated cell death in the nervous system. PMID- 8714526 TI - Glycine and GABA receptors: molecular mechanisms controlling chloride ion flux. AB - We have been able to show that the three clearly identified atoms common to the inhibitory neurotransmitters glycine and GABA, that we previously hypothesized to serve as attachment points at the glycinergic and gabanergic receptor, can indeed interact through both electrostatic and hydrogen bonding to several amino acids, which have been identified in molecular biological investigations as both present and critical in the physiological functioning of key polypeptides common to these inhibitory receptors. In addition, amino acids also involved in stabilizing the interaction between the antagonists strychnine and R5135 at the glycinergic and gabanergic receptors, respectively, have been shown to fit our complex model. We identify in detail molecular mechanisms to explain how glycine and GABA initiate chloride ion movement from extraneuronal fluid in the synaptic cleft to intraneuronal volume. In addition, we also identify the molecular mechanisms involved in the blocking of chloride ion movement by strychnine at the glycinergic receptor and by R5135 at the gabanergic receptor. We also present two computer-generated color prints, one for the glycine receptor and one for the GABA receptor, which show the quantum mechanically geometry optimized complex formed between receptor side chains, i.e., the part of the amino acids in the polypeptide that interacts with the zwitterionic inhibitory neurotransmitters. These computer-generated color figures also show a) the important electrostatic and hydrogen bonding in these interactions, b) a van der Waals model of this complex to illustrate that no steric repulsions exist, and c) the molecular electrostatic potential energy map showing the electrostatic potentials of neurotransmitter bound to the receptor model. Finally, we show with computer calculations that the pseudo-rings, formed between the positive quanidinium group in arginine and one of the oxygen atoms in the carboxyl group in both glycine or GABA, result in a positive planar region which appears to be involved in a charge transfer complex with aromatic benzene groups in amino acids such as phenylalanine and tryosine. PMID- 8714528 TI - F-wave in patients with hemifacial spasm: observations during microvascular decompression operations. AB - In patients with hemifacial spasm (HFS), the spasm is due to cross compression of the facial nerve by a blood vessel. There are currently two hypotheses how the cross compression can cause HFS: 1. the spasm is caused by ephaptic transmission and hyperexcitability at the site of compression; and 2. the spasm is caused by hyperexcitability in the facial motonucleus. In peripheral nerves, F-waves, which result from the backfiring of antidromically activated anterior horn cells, have been proposed as indices of anterior horn cell excitability. Enhancement of the F waves in facial muscles also indicates increased excitability of the facial motonucleus. On the other hand, abnormal muscle response (AMR), which can be elicited by stimulating one branch of the facial nerve and recording electromyographically from muscles innervated by other branches of the facial nerve, is specific for patients with HFS. We have therefore measured the AMRs and the F-waves in the facial muscle of HFS patients under anesthesia in order to investigate the excitability of the facial motonucleus. We obtained facial nerve evoked electromyograms from 14 HFS patients during microvascular decompression (MVD) operation. The F-waves, obtained with surface electrodes from the mentalis muscle, were defined as the second response after the M-wave. The F-waves in facial muscles cannot usually be elicited during surgical anesthesia using inhalation anesthetics. However, the F-waves were elicited on the spasm side in 10 out of 14 patients with HFS and the F-waves disappeared after MVD under anesthesia, as the early responses (R1) of the blink reflex were elicited on the spasm side before MVD under anesthesia. The F-waves elicited during anesthesia were suppressed significantly, compared with those before MVD. These results suggest that excitability in facial motonucleus increased on the spasm side. PMID- 8714529 TI - Vision in dementia: contrast effects. AB - Contrast sensitivity has been shown to be affected in Alzheimer's disease (Ad). We investigated low contrast acuity and contrast sensitivity using clinical test charts in this patient population. Additionally, we tested patients with vascular dementia (vd) and mixed dementia (md), (Alzheimer' with vascular dementia). Contrast sensitivity was assessed using the Vistech VCTS 6500 test chart. Low contrast acuity was measured using the Regan charts at four contrast levels (96%, 50%, 25% and 11%). The patient population consisted of 19 Ad patients, 9 vd patients and 10 md patients. Reduction in acuity was found with contrast level in all cases. Regression lines were fit to the data and statistical analysis was performed. We did not find a statistically significant difference between the Ad and vd or md groups. We did, however, find a difference between the vd and md groups. We did find reduction in contrast sensitivity at all spatial frequencies when compared to the elderly normal. Correspondingly, we found a significant difference in acuity when compared with normal data at the four contrast levels tested. Acuity is reduced with contrast in all patient groups. Our contrast sensitivity results are similar to those reported in the literature. This study points out the importance of using simple clinical test charts and further underscores the idea that there is a primary visual deficit in Ad. PMID- 8714530 TI - A reproducible assay of polymerase chain reaction to detect trinucleotide repeat expansion of Huntington's disease and senile chorea. AB - A simple and reproducible method of polymerase chain reaction (PCR) assay was established to detect trinucleotide repeat expansion for Huntington's disease (HD) using a new DNA polymerase and buffer system. The system consists of an extremely heat stable DNA polymerase (Pfu), and a buffer supplemented with ammonium sulfate and dimethyl sulfoxide. Previous methods to amplify expanded alleles for HD have been very complex in PCR conditions, but the reproducibility was sometimes very low because of repetitive sequences around the primer sequences. With the present method, strong bands for the disease alleles were reproducibly visible in a conventional agarose gel stained with ethidium bromide without using isotopes. Three cases with sporadic HD and a case with senile chorea showed expanded alleles for HD with smaller sizes of the expansion than cases with typical HD. These results showed that the present method provides a simple and reproducible way to detect HD allele, and some cases with sporadic HD and senile chorea had expanded HD alleles. PMID- 8714531 TI - Rejection of the 'filtered noise' hypothesis to explain the variability of transcranial Doppler signals: a comparison of original TCD data with Gaussian scaled phase randomized surrogate data sets. AB - Until the last few years the correlation dimension (D2) or the Lyapunow exponent were the two dominant mathematical methods which were applied to identify possible chaotic behavior in biological systems. Detection of deterministic chaos is important, because it suggests that a relatively simple nonlinear model might explain the data. It was however discovered that these methods could give rise to an erroneous detection of chaos. For this reason a new method was proposed in which the originally measured data set was directly compared with a computer generated 'surrogate' data set with exactly the same linear correlations as the original. The basic idea is then to compute a nonlinear statistic for the original data and for each of the surrogate data sets. In principle any statistic can be used. We used the correlation dimension (D2), which measures the complexity of a time series. In this study we applied this surrogate method to estimate whether the variability of the transcranial Doppler (TCD) waveforms is the result of nonlinearity or not. From 10 healthy volunteers, left middle cerebral artery (MCA) blood flow velocities were measured by TCD examinations. An artifact free epoch of each TCD was used for analysis. From each original data set 50 surrogate data sets were constructed using the Gaussian-scaled phase randomized Fourier transform. For both the original and the surrogate data sets the D2 was measured. The D2 values of the original TCD waveforms differed significantly from the mean D2 of the surrogate data sets. Therefore the null hypothesis, which stated that the original TCD time series arise from filtered noise, is rejected and nonlinearity is detected. The clinical significance and implications are discussed. PMID- 8714532 TI - Variability in occurrence of embolic signals in long term transcranial Doppler recordings. AB - Albeit still unproven, it is supposed that the presence and number of asymptomatic circulating cerebral microemboli detected by transcranial Doppler ultrasound (TCD) may be an indicator of stroke risk. Little research has been done to assess the reproducibility of these data and the required time for recording. We examined one middle cerebral artery in 7 subjects with carotid, aortic or cardiac embolic source for 24 h by TCD. Analysis for embolic signals was done off line completely blinded to the diagnosis and the time of the day during the recording. Embolic signals were found in all 7 subjects varying from 0 per h to 13 per h. Embolic signals occurred throughout the day with a nonsignificant tendency towards higher values in the early morning hours. Half an hour recording would not have been suitable to rule out or to confirm the presence of embolic signals as there were too many gaps between embolic signals of more than 1/2 h. One hour is the required minimum. Concerning the number of embolic signals, even a recording of four hours yielded variabilities of 0.25 and 8 embolic signals per hour in the same person. We recommend to perform follow-up studies the same time of the day. In patients with a low number of embolic signals longer recordings or a lower detection threshold with a higher number of detected embolic signals are necessary to compare frequencies of embolic signals. PMID- 8714533 TI - Compton scatter correction in case of multiple crosstalks in SPECT imaging. AB - A strategy for Compton scatter correction in brain SPECT images was proposed recently. It assumes that two radioisotopes are used and that a significant portion of photons of one radioisotope (for example, Tc99m) spills over into the low energy acquisition window of the other radioisotope (for example, Tl201). We are extending this approach to cases of several radioisotopes with mutual, multiple and significant photon spillover. In the example above, one may correct not only the Tl201 image but also the Tc99m image corrupted by the Compton scatter originating from the small component of high energy Tl201 photons. The proposed extension is applicable to other anatomical domains (cardiac imaging). PMID- 8714534 TI - Human cardiovascular evolution and intracranial saccular aneurysms: an hypothesis. AB - Current predominant theories for the pathogenesis of intracranial saccular aneurysms have a congenital or a degenerative basis. Complementing these theories, the predilection of intracranial aneurysms for the human species has been attributed to a variety of human neurovascular morphological traits, which however, on closer scrutiny are not unique to the human species. Human evolution to a bipedal primate has required significant adaptation in several organ systems, with significant consequent biological costs. We hypothesize that susceptibility to intracranial aneurysms is part of this latter cost of adopting an upright stance with respect to the neurovascular systems. Evidence supporting the above hypothesis, is presented. PMID- 8714535 TI - Management of unruptured cerebral aneurysms. AB - A series of 62 patients treated surgically for one or several unruptured intracranial aneurysms is reported. 83 aneurysms were treated in 65 operations. The main locations of the aneurysms were: MCA 35%, ICA (posterior communicating) 22%, carotido-ophthalmic segment 12%, carotid bifurcation 11%, anterior communicating artery 11%, verterbro basilar artery 5%. The circumstances of discovery were: incidental 28%, multiple aneurysm 22%, headache 18%, ischemic episode 9%, mass effect 8%, seizures 6%. Overall, 8% of these unruptured aneurysms were certainly symptomatic, 58% were certainly asymptomatic, and for 34% the relationship with the mode of discovery was uncertain. The overall outcome of surgery was: good recovery 94%, moderately disabled 1.5%, severely disabled 1.5%, and death 3%. The post-operative complications were related to surgical technique in 2 cases, to a severe atherosclerotic state of the ICA in 1 case, and to the general arteriopathy of the patient in 1 case. The discussion reviews in the literature the various arguments developed in favor of an active treatment of the unruptured cerebral aneurysms. Three arguments are proposed. 1. The overall severity of the aneurysm rupture, with a mortality rate over 60%. 2. The cumulative risk of rupture of an unruptured aneurysm, which may be high in young patients (from 16 to 30% lifetime risk). 3. The good outcome of the surgical treatment of the unruptured aneurysm (mortality rate under 4%, morbidity rate approximately 6%). The operative risk is higher for large or giant aneurysms, for a patient with a history of ischemic cerebrovascular accident as mode of discovery, for elderly patients with arteriosclerotic thickening of ICA wall and aneurysm neck. The decision to treat or not to treat may be easier (mass effect, multiple aneurysm, acute headache) or more difficult (chronic headache, hemorrhage of other origin, seizures, incidental discovery). The endovascular treatment with occlusion of the aneurysms sac by means of coils is more and more an alternative to surgical treatment, but requires a long follow-up to ensure the absence of reexpansion of the coil-embolized aneurysms. The screening for unruptured aneurysms, especially in cases with familial intracranial aneurysms is more and more often proposed. The authors' opinion now is surgical clipping of small and middle-sized aneurysms in young patients, without severe associated pathology, and clearly agreeing with surgery. The limit of age for surgery is usually 65 years except for those aneurysms discovered after a mass-effect. Elderly patients, giant aneurysms, patients with contra-indication for surgery, are proposed for endovascular treatment. PMID- 8714536 TI - Combination tissue plasminogen activator and ticlopidine therapy in a rabbit model of acute thromboembolic stroke. AB - The successful application of thrombolytic therapy to acute myocardial infarction has prompted a reinvestigation of thrombolytic therapy for acute stroke. However, an examination of safety and efficacy of thrombolytic therapy in acute thromboembolic stroke has precluded the entry of patients taking either antiplatelet or anticoagulant therapy. It was therefore of interest, in an established rabbit model of thromboembolic stroke, to examine the use of tissue plasminogen activator therapy in combination with ticlopidine treatment. Following ticlopidine administration (10 mg kg-1, i.v., daily for 5 days), rabbits (n = 7) were embolized by injecting a tin-laden clot into the internal carotid artery with clot placement confirmed by x-ray. Three hours later, t-PA was initiated as a square-wave pulse (6.3 mg kg-1 total dose, given as a 20% bolus, with the remainder administered over 2 h as a continuous infusion). The protocol was continued for a total of 7 h following embolization. Complete clot lysis was demonstrated in 6 of 7 animals. Brain infarct size (triphenyltetrazolium chloride staining) was 36.0 +/- 12.9% hemisphere (mean +/- SEM). Both clot lysis rate and infarct size were very similar to that previously seen following administration of t-PA alone (58% and 31.6 +/- 6.4% hemisphere, respectively) but in marked contrast to previous results seen with intravenous aspirin (no clot lysis). These results suggest that antiplatelet agents used clinically for stroke prophylaxis (aspirin or ticlopidine) may influence the success rate of thrombolysis following initiation of thrombolytic therapy for acute thromboembolic stroke. PMID- 8714537 TI - Endovascular treatment of pericallosal artery aneurysms. AB - Recent technical developments have made endovascular treatment using controlled detachable coils suitable for intracranial aneurysms. However, endovascular approach of the aneurysms of the pericallosal artery remains difficult because of their usual small size and their anatomical situation. PMID- 8714538 TI - Brain tissue oxygen pressure, carbon dioxide pressure and pH during ischemia. AB - In this case we evaluated brain tissue pO2, pCO2 and pH during ischemia and arterial oxygen desaturation (hypoxia). In both situations brain pO2 decreased. During ischemia, tissue pCO2 increased while pH decreased; but both pCO2 and pH were stable during hypoxia. These results suggest that brain tissue pO2, pCO2 and pH measures provide information on tissue perfusion and oxygen availability during ischemia and hypoxia. PMID- 8714539 TI - The fine structure of large dense-core organelles in human locus coeruleus neurons. AB - Protein bodies, the characteristic spherical organelles present in human monoamine neurons, have been shown in previous electron microscope studies to originate as dense bodies in mitochondria. This study was designed to investigate the presence of catecholamine reaction products in the dense bodies of locus coeruleus neurons, in frozen fresh post-mortem brain tissue with the use of potassium permanganate (KMnO4) fixation. This fixation procedure forms a dense KMnO4/catecholamine reaction product, visible in the electron microscope, in the large dense-core vesicles of experimental animals. Our results demonstrate the localization of KMnO4 dense product in the cores of double membrane-bound spherical organelles, as well as in spherical structures in the matrix of typical mitochondria. No typical large dense-core vesicles were observed in these catecholamine neurons of the tissues studied. Our findings are consistent with the notion that altered mitochondria may contribute to the formation of a new type of large dense-core vesicle in the locus coeruleus neurons of man, which is probably an evolutionary adaptation of amine-storing organelles. PMID- 8714540 TI - Mechanism of heat shock protein 72 induction in primary cultured astrocytes after oxygen-glucose deprivation. AB - Induction of stress proteins is thought to be important in the protection of cells from a variety of environmental insults including heat, hypoxia and ischemia. The aim of this study was to compare the mechanism of induction of heat shock protein 72 (HSP72) in primary cultures of murine cortical astrocytes by heat and combined oxygen-glucose deprivation (OGD), a model of in vitro ischemia. 35S-methionine labeling and immunoblotting showed increased HSP72 synthesis and accumulation lasting for up to 24 h following heat or OGD. Heat induced a markedly greater amount of HSP72 mRNA and protein than did OGD. We then sought evidence of heat shock transcription factor-1 (HSF-1) activation. An increase in apparent molecular weight of nuclear HSF-1 after heat or OGD was observed, consistent with increased phosphorylation. To seek an explanation of the difference between heat and OGD as inducers of HSP72 we examined the binding activity of HSP72 + 73 to other proteins. More cellular protein was found to co immunoprecipitate with HSP72 + 73, and more HSP72 + 73 was found in the pellet fraction after heat shock compared to OGD. These results suggest that HSP72 induction is regulated in astrocytes at least in part at the level of HSF activation, by both heat and OGD. Reduced availability of free HSP72 + 73 in heated cells could be responsible for the greater magnitude of HSP72 induction after heat compared to OGD. PMID- 8714541 TI - Reversible focal ischemia model in the rat: correlation of somatosensory evoked potentials and cortical neuronal injury. AB - Somatosensory evoked potentials (SEPs) are valuable in experimental stroke studies, but only a few reports have dealt with small rodent models. Our experiments aimed to reproduce SEP monitoring during reversible middle cerebral artery occlusion (MCAo) in a surgical model of open craniotomy and vessel manipulations in the rat. The changes of median nerve SEPs were correlated to the degree of local cortical neuronal injury in the forelimb area. Based on a pilot group (n = 11) both parameters were examined at 2 h after reperfusion in a subsequent study with increase of MCAo time from 1 (n = 7) to 2 h (n = 7); 5 rats were sham operated. A significantly delayed and incomplete recovery of SEPs, expressed as a percentage of the mean preocclusion values was observed with 2 h compared to 1 h MCAo (20.8% versus 6.0% change in the latency and 59.5% versus 2.2% in the amplitude of the primary cortical response). The functional outcome was found to correlate consistently with the degree of neuronal damage in 1 and 2 h reversible MCAo (2.5% versus 79.2% severely damaged neuronal types III and IV). Our findings suggest, that 2 h MCAo followed by 2 h reperfusion produces a submaximal neuronal injury and partial electrical recovery in the periphery of the occluded vascular territory and could be therefore used as a reliable model for assessment of cerebroprotective drug efficacy. PMID- 8714542 TI - Time courses of changes in cerebral blood flow and blood-brain barrier integrity by focal proton radiation in the rat. AB - In order to know the pathophysiological mechanisms underlying radiation brain injury, cerebral blood flow and blood-brain barrier integrity were studied using N-isopropyl-p-[123l]iodoamphetamine (IMP) and [14C]-alpha-aminoisobutyric acid (AIB), respectively, in the rat focal proton radiation model (a single dose of 30 or 60 Gy radiation with 70 MeV proton beams). One, 2, 4, and 5.5 months after irradiation, [123l]IMP and [14C]AIB were intravenously injected and uptake of IMP and AIB in the cerebral cortex, striatum, hippocampus, and thalamus was measured. Significant decreases in IMP uptake were observed in the cerebral cortex and thalamus of the irradiated side at 4 and 5.5 months after 60 Gy irradiation; the effects at 5.5 months were more prominent than those at 4 months. AIB uptake markedly increased in all the brain regions of the irradiated side at 5.5 months after 60 Gy irradiation, and at 4 months, only in the hippocampus. The results suggest that there are dose- and time-dependent responses in radiation effects and regional differences in tissue vulnerability to radiation. Proton focal radiation model appears to be a useful model for studies of radiation brain injury in small animals such as rats. PMID- 8714543 TI - Brain effect of intracarotid administration of cisplatinum. AB - The antitumoral effect of Cisplatinum is well known, including its effect on brain tumors. With the purpose of establishing the effect of intraarterial cis dichlorodiammine platinum (CDDP) on the brain, Cisplatinum was injected in rabbits that were sacrificed at different intervals. The general clinical and neurological condition of the rabbits was unaffected, and histopathological examination of the rabbit's brain was normal. PMID- 8714545 TI - Introduction to special section on termination and group therapy. PMID- 8714544 TI - Interrelation between protein kinase C and nitric oxide in the development of vasospasm after subarachnoid hemorrhage. AB - This study was undertaken to investigate how protein kinase C (PKC) and nitric oxide (NO) interact to regulate the vascular tone, and how their interaction contributes to the development of vasospasm after subarachnoid hemorrhage (SAH). For these purposes, vasospasm was conducted with a canine model. We investigated the following subjects with arteries from intact animals and those from the SAH model, and compared the results between the two; tension at rest of isometric tension study, the effect of PKC inhibitors and of an inhibitor of NO synthesis on the tension at rest, and levels of guanosine 3',5'-cyclic monophosphate (cGMP) as an indicator of NO production. The tension at rest was enhanced in the artery from the SAH model compared to that from intact animals, and it was PKC dependent. Arteries from intact animals but not those from the SAH model developed tonic tensions by NO inhibitors, and these tonic tensions were suppressed by PKC inhibitors, and also by cGMP. An enzyme immunoassay revealed a decreased cGMP level in the SAH model. The evidence indicates that NO exerts a negative feedback control on PKC activation. Subarachnoid hemorrhage interferes with this feedback control, resulting in PKC-dependent enhanced vascular tone and vasospasm. PMID- 8714546 TI - Premature termination of group therapy: a clinical perspective. AB - Premature terminations are an inevitable if complex aspect of any therapy group, as they are of life. They can be destructive to the life of a group, and damaging to its members. It is possible, however, given proper preparation and thoughtful reflection, to make such premature endings effective. And sometimes, understanding the wish to leave may change potentially premature endings into turning points in a continuing therapy. I use two clinical events to illustrate these processes. Premature terminations are described as part of a continuum of endings that include dropouts, early endings, and good-enough terminations. PMID- 8714547 TI - The process of termination in long-term psychoanalytic group therapy. AB - Termination is a complex phase in the total therapeutic process. Criteria for termination of long-term analytic group therapy encompass both intrapsychic and behavioral changes. One of the useful ways of conceptualizing the termination phase is in terms of the issues of separation-individuation. Three different kinds of termination that can occur and the techniques for working with each are described. Multiple transferences are worked with in the long-term analytic group and both real and transferential losses acknowledged. The ability to leave group therapy with a stronger, better defined sense of self is one of the goals of therapy. PMID- 8714548 TI - Time-limited group psychotherapy. AB - The presence of a time limit shapes the entire therapeutic process and culminates in the management of termination. This paper outlines a protocol for dealing with termination in time-limited psychotherapy groups: (1) Select suitable members with similar capacity to use a time-limited format; (2) use a closed group format; (3) clarify the time boundaries at the beginning; (4) reinforce the time frame during the last half of the group; (5) provide focused interventions; (6) forestall premature termination; (7) reinforce termination themes of deprivation, resentment and anger, rejection, grief and loss, responsibility for self; (8) structure the final session; and (9) plan a 4-month follow-up visit to encourage ongoing application. These guidelines provide the therapist with a general structure to ensure that major termination themes are systematically addressed. They deal with powerful concepts of maturation and self-responsibility and incorporate the principal strategies that distinguish time-limited from time unlimited group psychotherapy. PMID- 8714549 TI - Limiting time-unlimited group psychotherapy. AB - For a number of reasons, we decided to bring three long-term, open-ended psychodynamic therapy groups to termination. We examine the process that began with the announcement of the change to a time-limited format, and concluded with the termination of the group. The transition had a powerful impact on the members and the group-as-a-whole. The therapists used supervision to deal with the intensity of the group process and the countertransference demands posed by the transition. Groups facing a similar change due to economic pressures may benefit from these observations. PMID- 8714550 TI - Termination in inpatient groups. AB - This article provides the therapist with a three-factor framework for understanding and constructively responding to members' termination in the inpatient group. The first factor is the context in which the inpatient group takes place. It will be argued that how the broader system defines and treats termination affects the group process at the time of termination. The second factor is the therapist's countertransference. Three especially common reactions (disappointment, abandonment anxiety, and powerlessness) will be described. The third factor is the characteristic need of inpatients at termination time. Each factor is described in terms of its potential impact on the termination process. Techniques and suggestions are offered that aim to mitigate the destructive forces of each factor and to enhance the therapeutic value of the termination event for the patient. PMID- 8714551 TI - Termination in group psychotherapy from the perspectives of contemporary object relations theory and self psychology. AB - Throughout the development of object relations theory and self psychology, the importance of attachments, availability, absences, limitations, and endings have been factors that have informed theory and practice. This article examines the multiple meanings and impacts of terminations in group psychotherapy at the intrapsychic, interpersonal, and group-as-a-whole levels, from the perspective of contemporary object relations theory and self psychology. Emphasis is placed on the initiation of a mourning process, with the dynamics of guilt, shame, and aggression toward objects who are separate or inaccessible, and the experience of a shortcoming or absence, associated with a loss of self-object oneness, soothing, and idealization from the narcissistic sector of the personality. Therapeutic opportunities afforded by terminations and the importance of resolving countertransference reactions are highlighted. PMID- 8714552 TI - Confidentiality in group psychotherapy: empirical findings and the law. AB - Empirical research suggests that group therapists generally discuss with prospective clients the importance of maintaining confidentiality, but are unlikely to inform them of the significant potential for violations of confidentiality. Therapists believe information about the risk of unauthorized disclosures will reduce the number of patients willing to enter group therapy and will inhibit the therapeutic dialogue. Therapists' failure to provide information sufficient to obtain informed consent, however, produces serious ethical problems and potential legal problems as well. The law of informed consent varies in different jurisdictions such that identical factual scenarios could produce different legal outcomes depending on the jurisdiction in which the case occurs. In spite of the proliferation of group interventions, empirical studies of confidentiality in group therapy have lagged behind similar research in individual psychotherapy. PMID- 8714554 TI - Response to group-as-a-whole article and discussion. PMID- 8714553 TI - Mitigating the power imbalance in supervisee/supervisor co-leadership teams. PMID- 8714555 TI - Spontaneous firing of NG108-15 cells induced by transient exposure to ammonium chloride. AB - 1. We report that NG108-15 (neuroblastoma x glioma) cells differentiated in defined serum-free media are capable of exhibiting stable automaticity (the spontaneous occurrence of regenerative action potentials) following exposure to extracellular perfusates containing NH4Cl. 2. Membrane depolarization (4-5 mV) concomitant with an increased pHi during NH4Cl exposure are followed by hyperpolarization (5-7 mV), sub-threshold oscillations, and spontaneous firing after the removal of NH4Cl. 3. Cells cultured in 10% serum did not exhibit automaticity. Cells cultured in serum-free media are twice as likely to show automaticity as those cultured in reduced (1.5%) serum media. 4. We have examined factors that contribute to the events following NH4Cl exposure, namely, membrane depolarization and hyperpolarization, subthreshold oscillations, and automaticity. The inward currents activated at more negative potentials and the ionic currents associated with pronounced afterhyperpolarization in NG108-15 cells cultured in serum-free media provide a basis for the repetitive activity in general and automaticity in particular. PMID- 8714556 TI - Aging does not affect steady-state expression of the Na+/Ca2+ exchanger in rat brain. AB - 1. Steady-state protein and mRNA levels of the Na+/Ca2+ exchanger were studied in old (24-month) and young (3-month) brains of male Fischer 344 rats by Western and Northern analysis. 2. Northern analysis with a cDNA proble for the Na+/Ca2+ exchanger amplified from human brain RNA indicated the presence of two transcripts for the Na+/Ca2+ exchanger (6 and 16 kb). Both transcripts were present in similar abundance in the cerebrum and hippocampus. In the cerebellum the 6-kb transcript predominated. The cerebellum had the highest overall level of expression. There were no significant age-related effects seen on the level of expression of either transcript in each of the brain areas tested. 3. Western analysis of plasma membrane vesicles purified from cerebral cortex identified a single protein of 116 kDa. Consistent with the Northern analysis, no age-related effect on protein levels was seen. 4. The mechanisms underlying altered Na+/Ca2+ exchange activity in aging rat brain (Michaelis, 1989) most likely do not involve changes in gene expression and are therefore more likely to represent posttranslation modifications of the protein. PMID- 8714558 TI - Neural control of the expression of a Ca(2+)-activated K+ channel involved in the induction of myotonic-like characteristics. AB - 1. Expression of the apamin-sensitive K+ channel (SK+) in rat skeletal muscle is neurally regulated. The regulatory effect of the nerve over the expression of some muscle ion channels has been attributed to the electrical activity triggered by the nerve and/or to a trophic effect of some molecules transported from the soma to the axonal endings. 2. SK+ channels apparently are involved in myotonic dystrophy (MD), therefore understanding the factors that regulate their expression may ultimately have important clinical relevance. 3. To establish if axoplasmic transport is involved in this process, we used two experimental approaches in adult rats: (a) Both sciatic nerves were severed, leaving a short or a long nerve stump attached to the anterior tibialis (AT). (b) Colchicine or vinblastine (VBL), two axonal transport blockers of different potencies, was applied on one leg to the sciatic nerve. To determine whether electrical activity affects the expression of SK+ channels, denervated AT were directly stimulated. The corresponding contralateral muscles were used as controls. PMID- 8714557 TI - Diurnal variation of corticotropin-releasing factor binding sites in the rat brain and pituitary. AB - 1. Corticotropin-releasing factor (CRF) is thought to be involved in the regulation of the diurnal activity of the hypothalamus-pituitary-adrenal (HPA) axis and to act as a neurotransmitter in the brain. To date it is unknown whether the binding sites of the central CRF system are subject to diurnal variations. 2. We measured the number of CRF binding sites over the course of a complete 24-hr light-dark cycle in the pituitary, amygdala, bed nucleus of the stria terminalis (BNST), cingulate cortex, visceral cortex, paraventricular nucleus of the hypothalamus, hippocampus, and locus ceruleus of rats by in vitro receptor autoradiography with iodinated ovine CRF. A 24-hr time course was also established for plasma CRF and corticosterone. 3. The diurnal pattern of plasma CRF does not correlate with the pattern of plasma corticosterone. Within the brain, CRF binding in the basolateral nucleus of the amygdala showed a U-shaped curve with maximum levels in the morning and a wide hallow between 1500 and 0100. A biphasic profile with a small depression in the afternoon and a more pronounced depression in the second half of the activity period is characteristic for the other brain areas and the pituitary. The profile for the pituitary correlates with those for the BNST and the area of the locus ceruleus. Furthermore, the diurnal pattern of CRF binding sites in the BNST correlates with that of the hippocampus, and the daytime pattern of the visceral cortex is similar to that of both the hippocampus and the BNST. 4. Since the CRF-binding profiles in the brain and the pituitary clearly differ from the profiles of both plasma CRF and corticosterone, one may assume that the diurnal pattern of central CRF binding sites is not directly coupled to the activity of the HPA axis. PMID- 8714559 TI - The PH domain of Ras-GAP is sufficient for in vitro binding to beta gamma subunits of heterotrimeric G proteins. AB - 1. The noncatalytic domain of Ras-GAP can affect signaling through G protein coupled receptors by a poorly understood mechanism. 2. In this study, fusion proteins containing elements of the noncatalytic domain of ras-GAP were examined for their ability to bind beta gamma subunits of heterotrimeric G proteins and phosphotyrosine-containing polypeptides. 3. Our results demonstrate that purified beta gamma dimers associated with bacterially expressed GAP proteins and that this association does not require SH2 or SH3 domains but is dependent on the presence of the GAP pleckstrin-homology (PH) domain. In contrast, only the SH2 domains are necessary for binding to tyrosine phosphorylated proteins. 4. These findings raise the possibility that heterotrimeric G proteins might affect functioning of ras-like proteins through beta gamma subunits acting on their regulatory molecules. PMID- 8714560 TI - The 21-aminosteroid U-74389F increases the number of glial fibrillary acidic protein-expressing astrocytes in the spinal cord of control and Wobbler mice. AB - 1. Wobbler mice suffer an autosomal recessive mutation producing severe motoneuron degeneration and dense astrogliosis, with increased levels of glial fibrillary acidic protein (GFAP) in the spinal cord and brain stem. They have been considered animal models of amyotrophic lateral sclerosis and infantile spinal muscular atrophy. 2. Using Wobbler mice and normal littermates, we investigated the effects of the membrane-active steroid Lazaroid U-74389F on the number of GFAP-expressing astrocytes and glucocorticoid receptors (GR). Lazaroids are inhibitors of oxygen radical-induced lipid peroxidation, and proved beneficial in cases of CNS injury and ischemia. 3. Four days after pellet implantation of U-74389F into Wobbler mice, hyperplasia and hypertophy of GFAP expressing astrocytes were apparent in the spinal cord ventral and dorsal horn, areas showing already intense astrogliosis in untreated Wobbler mice. In control mice, U-74389F also produced astrocyte hyperplasia and hypertophy in the dorsal horn and hyperplasia in the ventral-lateral funiculi of the cord. 4. Given in vivo U-74389F did not change GR in spinal cord of Wobbler or control mice, in line with the concept that it is active in membranes but does not bind to GR. Besides, U-74390F did not compete for [3H]dexamethasone binding when added in vitro. 5. The results suggest that stimulation of proliferation and size of GFAP expressing astrocytes by U-74389F may be a novel mechanism of action of this compound. The Wobbler mouse may be a valuable animal model for further pharmacological testing of glucocorticoid and nonglucocorticoid steroids in neurodegenerative diseases. PMID- 8714561 TI - Cells expressing preproenkephalin mRNA in the rat pineal gland are not serotonin producing pinealocytes: evidence using in situ hybridization combined with immunocytochemistry for serotonin. AB - 1. Preproenkephalin (PPEnk) mRNA expressing cells have been identified in rat pineal gland using radioactive in situ hybridization histochemistry. 2. Approximately 7% of the cells in the pineal gland (7.5 +/- 0.86, mean +/- 95% CI) express PPEnk mRNA. These cells are distributed throughout the pineal as either scattered single cells or small groups of cells with large round or oval nuclei. 3. Using in situ hybridization combined with ABC immunocytochemistry for serotonin (5-HT) in the same pineal sections, the PPEnk mRNA labeling cells are found not to be serotonin-immunoreactive cells. These data indicate that the PPEnk mRNA is expressed in a certain discrete subpopulation of cells in the rat pineal gland and these cells are not serotonin-producing pinealocytes. 4. The physiologic role of PPEnk-derived peptides in the pineal remains unknown. It is possible that these peptides either are synthesized and secreted as hormones or act as pineal paracrine signals. PMID- 8714562 TI - [Diagnosis of laryngeal cancer]. AB - The basic methods of larynx cancer diagnosis were presented. It included subjective and objective examination, and laboratory examination. The importance of detailed history of cases determined by the tumor localization was underlined. Indirect and direct laryngoscopy have not lost it importance despite the development of modern methods of larynx cancer diagnosis. Microlaryngoscopy provides the most valuable information about the localization and advancement of neoplasmatic change. Its also allows for precise taking of specimen for histopathological examination. Radiological method in larynx cancer diagnosis were also taken into consideration particularly laryngotomography which allows for evaluation of subglottic region, ventrical and recesus piriformis. The histological examination remains a decisive factor in the surgery of larynx cancer. Great diagnostic possibilities of neck lymphatic system and larynx are being provided by the ultrasonographic examinations, computer tomography and magnetic tomography of nuclear resonance which allow for detailed evaluation of neoplasm extend and appearance of recurrence and metastases. The greatest responsibility in diagnosis of larynx cancer is put on a doctor who first advised the patient. PMID- 8714564 TI - [Why is it worth to take care of IGA?]. AB - The protective role of IgA against infections of the upper respiratory tract is described. The action of IgA formation and the possibility of its stimulation by applying oral vaccines is presented. PMID- 8714563 TI - [Estimation of aromatic DNA adduct levels in laryngeal tumors in relation to cancer staging]. AB - Genotoxical agents as PAH, nitrosamines, aromatic amines located in tobacco smoke are responsible for disorders of structure and function of DNA chromosomes, proteins and also initiation of carcinogenesis. DNA adducts are recognized as measure of the biological effective dose of exposure to environmental genotoxicant. So far most studies on DNA adducts after human exposure to tobacco smoke have ben carried out on white blood cells, lungs and oral cavities tissue. The aim of this work was estimation of relationship between state of neoplastic disease (TNM system) and number of aromatic DNA adducts. The subject were 37 patients with primary larynx tumor. In every case histopathological investigation revealed squamous cell carcinoma. There were 33 total and 4 partial laryngectomies performed. From tumour and nontumour larynx tissue DNA was isolated. Analysis of DNA adducts was performed by the 32P-postlabelling method. The results were characterized by individual differentiation. The highest level of DNA adducts was found in larynx tumour cells. In case of strong smokers (30-40 cigarettes per day) the level of aromatic adducts was higher then in non- or ex smokers. In both tissues (tumour and non-tumour) the highest level of aromatic adducts was in T3 stage, the lowes in cause of T2 stage. In tissues with T4 stage the level of DNA adducts was intermediate. One has been observed decrease tendency in level of DNA adducts which is connected with increase of TNM stage. PMID- 8714565 TI - [Metastases in the prelaryngeal (delphian) lymph nodes in the course of the laryngeal cancer]. AB - The metastases in the pre-laryngeal (Delphian) lymph nodes in the course of laryngeal cancer are uncommon. Between 1984 and 1994, 1573 patients with the diagnosis of laryngeal carcinoma were operated at the Department of Otolaryngology of Academy of Medicine in Poznan (Poland). Total laryngectomies were performed in 1069 cases and partial laryngectomies in 504 cases. Pre laryngeal lymph nodes were identified in 109 patients. Histologically, metastases were found in 8 cases (7.48%). The prognosis of the patients with metastases in these lymph nodes is poor; only 2 patients are alive and free of neoplastic disease. PMID- 8714566 TI - [The endoscopic assessment of the pharyngeal opening of the auditory tube in recurrent exudative otitis media in children]. AB - In a group of 227 children endoscopic assessment of the auditory tube function and its pharyngeal opening anatomy was performed. The mentioned group was isolated from 2965 undergoing adenoidectomy. Preoperative endoscopy revealed impaired functioning of the auditory tube and also abnormal appearance of its pharyngeal opening in 71.3% of cases. Within the group adenoidectomy wasn't effective in 32 patients. PMID- 8714567 TI - [The usefulness of cupular-otolith integration examination in postrotary nystagmus for estimation of otolith function]. AB - The aim of work was the influence of otolith irritation on duration of postrotatory nystagmus. The otolith excitation was performed by the head bend on the shoulder homolateral or heterolateral for the direction of postrotaroty nystgamus. The results show the difference in time duration of nystagmus in these two manoeuvres. In author opinion this differentiation makes possible separate ototlih examination. PMID- 8714568 TI - [Hearing impairment in children with Down's syndrome]. AB - The purpose of this study was the examination of 60 children with Down's syndrome with particular attention paid to the organ of hearing. The investigation included anamnesis concerning the risk factors of hearing loss, psychomotoric development, otolaryngological examination, pure tone, impedance and speech audiometry. The main conclusion is: hearing impairment is more frequent in children with Down's syndrome (82.8%) than in normal population (0.5-2%). The type of hearing impairment is mainly conductive and mild degree. PMID- 8714569 TI - [Original technical inventions in ENG]. AB - Authors showed own original vestibular testing instruments. The instruments have been used in the Department of Otolaryngology of the Medical Academy in Bialystok for many months. PMID- 8714570 TI - [Otogenic sigmoid sinus thrombosis in the course of chronic otitis media]. AB - A four cases of sigmoid sinus thrombosis due to chronic exacerbated otitis media were presented. In all patients surgical intervention involved a radical mastoidectomy and exposure of the sinus. In two cases sigmoid sinus was incised and the internal jugular vein ligated. In the two others, sinus was only needled. A high dosages of antibiotics were administered parenterally. All patients were discharged in good state of heals. PMID- 8714571 TI - [Aspergillosis in the anterior ethmoid: case report]. AB - The case of aspergillosis in the anterior ethmoid and maxillary sinus was described. The clinical course, diagnostic and treatment were discussed. PMID- 8714572 TI - [Granular cell myoblastoma (Abrikossoff tumor) of the larynx]. AB - Authors described a case of a 62 year old woman with non-typical laryngeal seat of granular-cell myoblastoma. In this case Abrikossoff tumor was located within the vestibular wall of the larynx (in the aryepiglottic and false folds). It was removed by laryngofissurae. During eight years of post-operative period no symptoms of recurrence were observed. While discussing literature of granular cell myoblastoma the authors draw attention to histopathology, clinical course of this disease and stress the rarity of occurrence of Abrikossoff tumor in the larynx. PMID- 8714573 TI - [Pneumocephalus following facial injury]. AB - Pneumocephalus or intracranial air its commonly detected following serious head injury and posterior fossa surgery. In laryngology the frequent reason of its appearance is also herniation of air through a break in the wall of skull due to advanced benign and malignant tumors. The pathophysiology involves the presence of craniodural fistula allowing ingress of air. A ballvalve mechanism may allow air to enter not exit the cranium or like in turned upside down bottle leak permit air entrance as fluid leaves the intracranial space. The presence of air is usually asymptomatic but caries a potential risk of increased intracranial pressure or meningitis which require an immediate therapy. Although various ways of treatment were reported prevention and early diagnosis are the most important. We report 3 cases of cerebral air that occurred secondary to the head trauma. The diagnosis every time promptly confirmed by skull roentgenogram or CT. PMID- 8714575 TI - [A new biopsy needle in the laryngological diagnosis]. AB - An own constructed biopsy needle is described. The technique is applied in the deep tissue pathological lesions (neoplastic, mycotical and for bacterial infiltrations). The mean application is of endophitic infiltration diagnosis of the larynx. The authors stress the fact, that the needle seems to be useful in the pathomorphological diagnosis of the neck and breast tumors. PMID- 8714574 TI - [Adenoid cystic carcinoma of the larynx, trachea and thyroid]. AB - The aim of this article was to describe rarely occurred carcinoma adenoides cysticum located in the larynx, trachea and thyroid. Epidemiology, clinical course and medical therapy was demonstrated. In reported case concerning 65-year old woman early bilateral paresis of vocal folds was observed as results of the perineural infiltration. That caused sudden laryngeal dyspnea. It is important to pay attention on local extensiveness of neoplasm. Besides changes in larynx, neoplastic infiltration was found in trachea, preesophageal tissues and right lobe of thyroid. For above reasons there is no possible to exclude thyroid as a primary tumor of carcinoma adenoides cysticum. PMID- 8714576 TI - [Nineteenth century physicians against drum perforation. I. The method of "mechanical" closure of drum perforation]. AB - The trials of closure of drum perforation since a long are described. The first, who noticed drum perforation was Astley Paston Cooper. The first, who used artificial ear-drum was Marcus Banzer. The achievements of James Yearsley, Juliusz Erhard Camile Miot, Adam Politzer, Joseph Toynbee and others are presented. The achievements of Polish physicians: Teodor Heiman, Aleksander Zebrowski, Jan Szmurlo, Edmund Modrzejewski, Rafal Spira and others are depicted. The indications and contrinications for artificial ear-drum employment are also described. PMID- 8714577 TI - [The actuality of Jan Miolonski's pioneering ideas in otolaryngology]. PMID- 8714578 TI - [The prognostic value of electronystagmographic and electroencephalographic tests in patients in brain coma]. PMID- 8714579 TI - [European federation of audiological societies (EFAS). Goals and purposes of the organization]. PMID- 8714580 TI - [Obituary to Professor Leon Handzel, PhD]. PMID- 8714581 TI - Oral iron chelation. PMID- 8714582 TI - Hemoglobin switching and fetal hemoglobin reactivation. PMID- 8714583 TI - Reactivation of fetal hemoglobin in patients with beta-thalassemia. PMID- 8714584 TI - Towards gene therapy of hemoglobinopathies. PMID- 8714585 TI - Psychological and sociological aspects of the thalassemias. PMID- 8714586 TI - Clinical experience of management of thalassemia: the Sardinian experience. PMID- 8714587 TI - Current status of thalassemia and the sickle cell syndromes in Greece. PMID- 8714588 TI - Fluorogenic peptide substrates for assay of aspartyl proteinases. AB - Via a combination of chemical and enzymatic synthesis, new hexapeptide substrates convenient for use in activity assessment of several aspartyl proteinases- porcine pepsin, human pepsin, gastricsin, and cathepsin D--were prepared. These peptide derivatives, o-aminobenzoyl-Ala-Ala-Phe-Phe-Ala-Ala-p-nitroanilide and N (o-aminobenzoyl-Ala-Ala-Phe-Phe-Ala-Ala)-N'-2,4-dinitrophenyl ethylenediamine, contain a fluorescent o-aminobenzoyl moiety as well as p-nitroaniline or N-2,4 dinitrophenyl ethylenediamine--the groups that cause fluorescence quenching. Aspartyl proteinases hydrolyze the Phe-Phe peptide bond in the substrates, which diminishes quenching due to separation of the fluorescent and quenching moieties and leads to an increase in the fluorescence intensity of o-aminobenzoyl residue. Abz-Ala-Ala-Phe-Phe-Ala-Ala-Ded, being fairly well hydrolyzed by HIV proteinase, might be used for assay of this enzyme. PMID- 8714589 TI - A mass spectrometry-based approach for probing enzyme active sites: identification of Glu 127 in Cellulomonas fimi exoglycanase as the residue modified by N-bromoacetyl cellobiosylamine. AB - We have identified the residue in Cellulomonas fimi exoglycanase modified by N bromoacetyl cellobiosylamine as Glu 127 using a new combination of experimental approaches. The enzyme was quantitatively inhibited with the affinity label N bromoacetyl cellobiosylamine and cleaved with pepsin. The N-acetyl cellobiosylamine-modified peptide was identified by comparative peptide mapping of the digests derived from labeled and unlabeled proteins by reverse-phase high performance liquid chromatography connected online to an electrospray ionization mass spectrometer. The modified residue in the labeled peptide was determined by using a novel protein sequencing chemistry which is based on monitoring the amino acid derivatives released by stepwise peptide degradation using electrospray ionization mass spectrometry. Tandem mass spectrometry was used for further structural characterization of the cleaved residue. We show that the residue modified by N-bromoacetyl cellobiosylamine is Glu 127. This residue has been identified previously as the acid-base catalyst by using a combination of mutagenic and kinetic analyses. Our results therefore demonstrate the usefulness of this type of affinity label in identifying important catalytic residues in glycosidases and suggest that this new experimental approach can be applied generally to any labeled protein in which the mass of the label is known and thus represents an alternative approach to the current methods used to identify labeled residues within proteins. PMID- 8714590 TI - Combinatorial peptide libraries: robotic synthesis and analysis by nuclear magnetic resonance, mass spectrometry, tandem mass spectrometry, and high performance capillary electrophoresis techniques. AB - Combinatorial peptide libraries are a new source of compounds from which a large number of pharmacological leads will emerge in the next few years. A large body of literature shows that this approach is of considerable interest in many areas of biological sciences for the search of enzyme substrates and inhibitors, of receptor agonists or antagonists, or of antigen sequences. Nevertheless, the analytical investigation of such complex mixtures as libraries which contain up to millions of individual molecules is still poorly documented. In this work, we present analytical solutions for their characterization. NMR and tandem mass spectrometry (MS/MS) can provide an in deep description of any type of combinatorial libraries, while MS and high-performance capillary electrophoresis can bring a rapid and overall information at the routine level, sufficient to ensure a first assessment of their composition. MS in the fast atom bombardment mode was used to describe the libraries O1X2X3X4X5 or O1X2X3X4 (Oi and Xi are fixed and random residue in position i, respectively). Advantage was taken of the high proton affinity of arginine and of its induction of charge remote fragmentation to interpret the MS spectra of whole libraries and neutral losses (MS/MS) in the model sublibraries ArgGlyX3X4 and NipValX3X4 (Nip,4 nitrophenylalanine). Two-dimensional NMR allowed the incorporation of the individual residues during synthesis to be tested in 24 sublibraries O1X2X3X4. While from the pharmacological point of view, impressive discoveries made with combinatorial peptide libraries have already been reported, our results show that they should be complemented by appropriate analytical tools, crucial for the proper characterization and exploitation of these libraries. PMID- 8714591 TI - Water activity by 1H nuclear magnetic resonance spectroscopy: application to the study of water exchange in biphasic media. AB - The water activity (alpha w) of the liquid phase is investigated by means of 1H NMR for both monophasic and biphasic systems. The chemical shift or the area of the signal of the hydroxylic hydrogens is compared to calibration curves obtained from mixtures equilibrated at different water activities, thus allowing determination of the alpha w of the system. The chemical shift varies linearly as a function of the alpha w of the system. Through simple calculation the hydroxylic hydrogen concentration obtained from the area gives the water concentration in the medium; the sorption curve is thus obtained through NMR. The application of this method to a biphasic system composed of a liquid phase (an equimolar mixture of acid-alcohol) and a solid phase (a lipase), equilibrated separately at two different water activities, gives information on the water exchange between both phases during the lag phase of the esterification reaction. Light and slow water exchanges are observed from the solid phase at high alpha w toward the liquid phase at low alpha w. The lag phase of the esterification reaction is too short for the water activity equilibrium to be reached before the reaction starts. PMID- 8714592 TI - Immobilized metal ion affinity capillary electrophoresis of proteins--a model for affinity capillary electrophoresis using soluble polymer-supported ligands. AB - An affinity capillary electrophoresis method has been developed that employs small ligands covalently bound to a replaceable soluble polymer matrix. The metal chelate iminodiacetate-Cu(II) coupled to polyethylene glycol was used as a model system and the interactions of different model proteins, namely ribonucleases, cytochromes c, chymotrypsin, and kallikrein, were investigated. The method allowed for easy determination of dissociation constants using a modified Langmuir adsorption isotherm equation which is applicable to interactions with fast on/off kinetics. It is shown that the general rules for protein interaction with matrix-bound metal chelates established for immobilized metal ion affinity chromatography (IMAC) are maintained in immobilized metal ion affinity capillary electrophoresis (IMACE). In contrast to gel electrophoresis, IMACE allowed for using similar conditions as in IMAC, especially regarding the high salt concentrations, usually employed with the latter technique. The usefulness of the method for quantification of low-affinity interactions and studying protein surface structure and structure/function relationship using the metal affinity is demonstrated. PMID- 8714594 TI - "Doublex" fluorescent DNA sequencing: two independent sequences obtained simultaneously in one reaction with internal labeling and unlabeled primers. AB - The novel "doublex" DNA sequencing technique that makes it possible to obtain simultaneously two independent sequences from one sequencing reaction with the use of unlabeled primers and internal labeling is described. The different sequencing products are labeled in parallel with fluorescein-15-dATP and Texas red-5-dCTP present in the same tube. The characteristics of T7 DNA polymerase are exploited to ensure that only either of the labeled dNTPs is incorporated into the corresponding sequencing products. Specificity of labeling is ensured by the selection of primers. One of the unlabeled primers is chosen to be followed by an "A," the other by a "C" to be incorporated immediately downstream from the primer binding site. The doublex sequencing technique is applicable to the simultaneous sequencing of either the same DNA template/strand or a mixture of different templates. Combinations of unlabeled and labeled primers in the same sequencing reaction are also possible. The two sequences can be determined in parallel and on-line in the same lanes of a gel with a novel automated DNA sequencer, which was previously described for use with labeled primers. PMID- 8714593 TI - Competition BIAcore for measuring true affinities: large differences from values determined from binding kinetics. AB - In attempting to use the BIAcore instrument for the determination of binding constants of several haptens or peptides to different antibodies by measuring on- and off-rates, we found that neither the absolute nor the relative values of the binding constants corresponded to the measurements in solution. Even at the lowest coupling densities useful for measurements, rebinding and bivalency effects offset the measurements by a factor of up to 500. We caution therefore about using on- and off-rates for the determination of absolute or even relative binding constants without controlling for rebinding and avidity effects. Instead, we show that binding constants in solution can be reproduced well by using on rate determinations of antibody preincubated with antigen, and we derive the conditions under which such an approach is valid. PMID- 8714595 TI - A general progress curve method for the kinetic analysis of suicide enzyme inhibitors. AB - The efficiency of suicide inhibitors is expressed in terms of the kinetic parameters KI and kinact and the partition ratio, which are commonly determined by the well-known dilution assay method. Progress curve analysis methods have been widely used in the determination of the kinetic parameters of active-site directed affinity labels and have also been applied to a few suicide inhibitors where the turnover rate of the regular substrate could be measured independently of the turnover rate for the inhibitor, when both the substrate and the inhibitor are present in the assay medium. However, the progress curve analysis method for suicide inhibitors has not been applied to the most common case where the progress curve is a result of the turnover of both substrate and inhibitor. In the present study we have attempted to apply this method to a well-characterized suicide inhibitor of dopamine beta-monooxygenase (EC 1.14.17.1) where the progress curve is a result of the turnover of both substrate and inhibitor. These results demonstrate that the kinetic constants determined by this method are highly reproducible and are also in excellent agreement with those previously determined by the dilution assay method. Efficiency and reproducibility, as well as the adaptability of commercially readily available curve-fitting programs such as Sigma Plot for routine use, are major advantages of this procedure. PMID- 8714597 TI - Electrophoresis of polyethylene glycols and related materials as sodium dodecyl sulfate complexes. AB - Polyethylene glycol (PEG) and polyethylene glycol derivatives are analyzed by a modification of the sodium dodecyl sulfate-polyacrylamide stacking gel electrophoresis procedure of Kurfurst. Gels using a discontinuous buffer system but which do not have a separate stacking gel are used without loss of resolution and with less tendency to form artifactual multiple or distorted bands. Examination of several commercial preparations of PEGs and PEG derivatives on such gels indicates heterogeneity other than the expected unimodal size distributions. SDS-gel electrophoresis of proteins or other materials in samples containing PEGs may yield gels with zones of contamination from the PEGs. Methods of reducing such contamination are suggested. PMID- 8714596 TI - Determination of benzidine, N-acetylbenzidine, and N,N'-diacetylbenzidine in human urine by capillary gas chromatography/negative ion chemical ionization mass spectrometry. AB - We report an isotope dilution mass spectrometry method using capillary gas chromatography/negative ion chemical ionization to quantitate urine concentrations of benzidine (BZ) and of its acetylated metabolites N acetylbenzidine (ABZ) and N,N'-diacetylbenzidine (DABZ). Urine samples were purified by solid-phase extraction columns, reduced with LiAlH4/THF, and derivatized with pentafluoropropionic anhydride. The derivatives were measured by selected ion monitoring relative to deuterium-labeled internal standards. A detection limit as low as 0.5, 0.8, and 1.5 ppt for BZ, ABZ, and DABZ, respectively, can easily be achieved. Urinary concentrations of ABZ substantially exceed those of either BZ or of DABZ in workers exposed to BZ or BZ-based dyes. This method has been successfully used to measure BZ, ABZ, and DABZ in 1.0-ml urine samples collected from workers involved in manufacturing BZ and BZ-based dyes. The method should be applicable to the measurement of other aromatic amines and their acetylated metabolites. PMID- 8714598 TI - Semiautomated method for the quantitation of plasma or sera androstenedione, testosterone, and dihydrotestosterone in population studies. AB - A method is presented that analyzes quantitatively and reproducibly the androgens testosterone, androstenedione, and dihydrotestosterone from human sera or plasma. The chromatographic separation step generates an unattended throughput of one preparative separation per hour. Controls are built into the method to account for changing chromatographic conditions that otherwise result in shifts in retention characteristics. Separation factors for the three androgens are as follows (mean +/- standard deviation): alpha = 1.23 +/- 0.011 between androstenedione and testosterone and alpha = 1.38 +/- 0.025 between testosterone and dihydrotestosterone. Sensitivities of the method are androstenedione 5 pg, testosterone 3 pg, and dihydrotestosterone 14 pg. A study of procedural losses associated with initial sample processing, a validation, and application to two sample sets which demonstrates the methods utility for the analysis of hypoandrogenic populations (postmenopausal women) and hyperandrogenic groups (prostate cancer patients) is also reported. The precision for replicate aliquots of control plasma is androstenedione and testosterone = 5-11% CV and dihydrotestosterone = 10-20% CV. PMID- 8714599 TI - Candidate reference method for determining serum cortisol based on isotope dilution-gas chromatography/mass spectrometry using heptafluorobutyrilation as derivatization method. AB - We present a new candidate reference method for determining cortisol in serum. The method is based on isotope dilution-gas chromatography/mass spectrometry and makes use of derivatization with heptafluorobutyric anhydride and selected ion monitoring at m/z 489 and 491. A detection limit of 0.08 pmol (30 pg) was achieved. Twenty-four structurally related steroids were tested for interference and found negative. Verification of the analytical quality specifications was done from measurement in two laboratories of the certified reference materials 192 and 193 of the Bureau Communautaire de Reference and a number of commercial quality control materials. The maximum systematic error was estimated to be 1.0% and the mean imprecision was 1.0%. The total error was lower than 2.05%. The method was applied for target-setting in external quality assessment and internal accuracy control and for measurement of patient samples. PMID- 8714600 TI - Polymerase chain reaction-based site-directed mutagenesis using magnetic beads. AB - Thyroglobulin double mutants with various substitutions were obtained with a new polymerase chain reaction-based mutagenesis technique. Maximum length of megaprimer and efficiency of mutagenesis were improved by purification of single stranded DNA, using the avidin-biotin interaction. This method might allow the construction of large libraries of DNA, mutated at different sites. PMID- 8714601 TI - Esterase coupled with the H2O2/horseradish peroxidase system triggers chemiluminescence from 2-methyl-1-propenylbenzoate: a potential analytical tool for esterase analysis. AB - The hydrolysis of 2-methyl-1-propenylbenzoate catalyzed by esterase produces 2 methyl-1-propenol, which can be subsequently oxidized by the H2O2/horseradish peroxidase (HRP) system to yield electronically excited triplet acetone. The level of luminescence elicited by this species is proportional to total esterase used, making it possible to determine as little as 2 pmol of esterase. Yet, its intensity can be enhanced several orders of magnitude by fluorescent acceptors like sodium 9,10-dibromoanthracene-2-sulfonate. The system works as a chemiluminescent reaction triggered by esterase and can be used to elaborate analytical assays to determine its activity. This chemiluminescence is also promoted by HRP conjugates instead of free HRP and, hence, this simple reaction system can also be used to develop sensitive chemiluminescent immunoassays based upon peroxidase activity. PMID- 8714602 TI - Methylglyoxal assay in cells as 2-methylquinoxaline using 1,2-diaminobenzene as derivatizing reagent. PMID- 8714603 TI - Quantitative reverse transcriptase-polymerase chain reaction of eukaryotic mRNA with the prokaryotic chloramphenicol acetyltransferase RNA as a universal, internal standard. PMID- 8714605 TI - An apparatus for immunoblotting using a magnetic rubber sheet. PMID- 8714604 TI - The histidine tail of recombinant DNA binding proteins may influence the quality of interaction with DNA. PMID- 8714606 TI - Effect of the concentration of 8-anilinonaphthalenesulfonic acid on the detection of folding intermediates in alpha-urease. PMID- 8714607 TI - Utilization of glass HPLC autosampler vials as cuvettes in an iodometric assay of hydroperoxides. PMID- 8714608 TI - Patient selection and predicting response to recombinant human erythropoietin in anemic cancer patients. PMID- 8714609 TI - Transfusion requirements in patients with malignancy. PMID- 8714610 TI - Mobilization of hematopoietic progenitor cells with Epoetin alfa. PMID- 8714611 TI - Recombinant human erythropoietin in combination with other hematopoietic cytokines in attenuating chemotherapy-induced multilineage myelosuppression: brief communication. PMID- 8714612 TI - Practice policies for surgical red blood cell transfusion: surgical transfusion policies. PMID- 8714613 TI - Current results and prospective trials of cladribine in chronic lymphocytic leukemia. PMID- 8714614 TI - The Scripps Clinic experience with cladribine (2-CdA) in the treatment of chronic lymphocytic leukemia. PMID- 8714615 TI - Cladribine in the treatment of low-grade non-Hodgkin's lymphoma. PMID- 8714616 TI - Cladribine for the treatment of indolent non-Hodgkin's lymphomas. PMID- 8714618 TI - Medical ethics, Islam and our society. PMID- 8714617 TI - Treatment of multiple sclerosis and other autoimmune diseases with cladribine. PMID- 8714619 TI - Erythrocyte glucose 6 phosphate dehydrogenase deficiency and neonatal jaundice. AB - Four years data from Special Care Baby Unit revealed neonatal jaundice (NNJ) as the commonest cause of hospitalization (1944 cases of NNJ out of 6454 admitted neonates). Majority (47.5%) of babies with NNJ presented between 4-7 days of birth. One hundred and sixty infants with NNJ were positive for Glucose 6 Phosphate dehydrogenase (G6PD) deficiency, of whom 153 were males and 7 females. Eighty five G6PD deficient babies required exchange transfusion and 23 developed bilirubin encephalopathy (BE) of which 7 died. PMID- 8714620 TI - Obstructed labour: the preventable factors. AB - Sixty-two cases of obstructed labour were reviewed during 6 months at a district head quarter hospital, Faisalabad to highlight the prevalence of obstructed labour and ascertain how to manage and possibly avoid this complication with special emphasis on preventable factors. There were 1584 deliveries during this period giving an incidence of obstructed labour as one in 25 deliveries. Five patients had rupture of uterus and there was one maternal death. PMID- 8714621 TI - A study on serum triglyceride and lipoproteins cholesterol in coronary heart disease patients. AB - Sera obtained from 42 patients of myocardial infarction, 28 of unstable angina and 25 healthy control subjects was assayed for total triglycerides, cholesterol, LDL cholesterol (LDL-C), HDL Cholesterol (HDL-C) and VLDL Cholesterol (VLDL-C). The level of cholesterol was also determined among the sub-fractions of HDL. The levels of total triglycerides, cholesterol, LDL-C and VLDL-C were significantly raised in both groups of patients. Total high density lipoprotein cholesterol was low in patients, which was highly significant in HDL2-C and not in HDL3-C. These results suggest that serum lipids are directly correlated with the severity of coronary heart disease (CHD). Total cholesterol when measured in major lipoprotein classes and further partitioned into subfractions, increase their sensitivity, specificity and hence their predictive value as sensitive markers for the assessment of CHD risk. PMID- 8714622 TI - Clinical features of infantile diarrhea associated with single or multiple enteric pathogens. AB - Clinical features of infantile diarrhea were studied among 603 infants from birth to 12 months of age to determine the predominant clinical feature(s) seen in infantile diarrhea associated with a specific enteric pathogen. Among the major clinical features, fever was most often seen in diarrhea due to Yersinia spp. (61.5%) followed by that in rotavirus (26.1%). Vomiting was mostly associated with Vibrio cholerae infection (90.9%) and shigellosis (64.6%). Dehydration was predominant in Vibrio cholerae (90.9%) and Salmonella (84.9%) infections. Bloody diarrhea was mostly due to Shigella infection (74.3%). As regards diarrhea with multiple pathogens, vomiting and dehydration were most frequent with Campylobacter+Enteropathogenic Escherichia coli (EPEC) (88.9% and 77.8%, respectively), while fever was more common with rotavirus+Shigella+Escherichia coli and rotavirus+Giardia. Infection with invasive organisms lead to vomiting, 4 10 stools per day and dehydration significantly more often as compared to infections with non-invasive organisms. Similarly more stools of patients infected with invasive organisms showed presence of blood and more than 5 leukocytes/HPF as compared to those infected with non-invasive organisms. PMID- 8714624 TI - A case of severe ischemic colitis caused by systemic lupus erythematosus. PMID- 8714623 TI - Prevalence of antibody to hepatitis C virus in Pakistani thalassaemics by particle agglutination test utilizing C 200 and C 22-3 viral antigen coated particles. AB - Exposure to hepatitis C virus (HCV) and its effect on ALT levels was studied in 35 transfusion dependent cases of thalassaemia major. Twenty-one (60%) cases were anti HCV positive and also showed raised Alanine Transaminase (ALT) levels. Of 14 anti HCV negative, Hepatitis B Surface Antigen (HBs Ag) negative seven showed raised ALT levels, indicating the chances of acute viraemia. Thus there is an urgent need to start anti HCV screening on all blood donations. PMID- 8714625 TI - Pneumonia caused by nocardia caviae in a healthy neonate. PMID- 8714626 TI - Primary amyloidosis of gut presenting with dysphagia. PMID- 8714627 TI - An update on hyperlipidemia and its management. PMID- 8714628 TI - Is yoghurt a source of gastroenteritis? PMID- 8714629 TI - A young boy with abdominal pain. PMID- 8714630 TI - Secnidazole in the treatment of giardiasis. PMID- 8714631 TI - Classifying illusory contours by means of orientation discrimination. AB - 1. Thresholds for discrimination of orientation were measured for solid and interrupted lines, and for illusory borders defined by offset abutting lines of a variety of configurations. 2. Illusory contours defined by orthogonal lines with a clear gap or overlap manifest excellent orientation discrimination even for very short exposure durations, and are robust to pattern motion and masking. The performance is almost as good as that for real lines. 3. When the contour is made up of abutting lines without gap or overlap, orientation discrimination is not nearly as good, and deteriorates further with short exposure duration, with target motion, and with masking. 4. The introduction of disparity in such illusory borders does not improve the discrimination of its orientation, so that concepts such as surface elaborations do not yet have to be invoked. 5. The results suggest that there is one class of illusory contour whose orientations can be discriminated by virtue of its delivering adequate stimuli to orientation selective units in primary visual cortex. For contours with a uniform space averaged luminance, the orientation discrimination is helped by the orthogonality of the generating lines, and this gives support to the concept that orthogonally oriented end-stopped neurons may participate in the orientation discrimination. PMID- 8714632 TI - Whole body withdrawal circuit and its involvement in the behavioral hierarchy of the mollusk Clione limacina. AB - 1. The behavioral repertoire of the holoplanktonic pteropod mollusk Clione limacina includes a few well-defined behaviors organized in a priority sequence. Whole body withdrawal takes precedence over slow swimming behavior, whereas feeding behavior is dominant over withdrawal. In this study a group of neurons is described in the pleural ganglia, which controls whole body withdrawal behavior in Clione. Each pleural withdrawal (Pl-W) neuron has a high threshold for spike generation and is capable of inducing whole body withdrawal in a semi-intact preparation: retraction of the body-tail, wings, and head. Each Pl-W neuron projects axons into the main central nerves and innervates all major regions of the body. 2. Stimulation of Pl-W neurons produces inhibitory inputs to swim motor neurons that terminate swimming activity in the preparation. In turn, Pl-W neurons receive inhibitory inputs from the cerebral neurons involved in the control of feeding behavior in Clione, neurons underlying extrusion of specialized prey capture appendages. Thus it appears that specific inhibitory connections between motor centers can explain the dominance of withdrawal behavior over slow swimming and feeding over withdrawal in Clione. PMID- 8714633 TI - Cerebral serotonergic neurons reciprocally modulate swim and withdrawal neural networks in the mollusk Clione limacina. AB - 1. A pair of serotonin-immunoreactive neurons has been identified in the cerebral ganglia of the pteropod mollusk Clione limacina, which produce coordinated, excitatory/inhibitory effects on neurons controlling two incompatible behaviors, swimming and whole body withdrawal. These cells were designated cerebral serotonergic ventral (Cr-SV) neurons. 2. Activation of Cr-SV neurons produces a prominent inhibition of the pleural withdrawal neurons, which have been previously shown to induce whole body withdrawal in Clione. In addition, the cerebral neurons produce weak excitatory inputs to swim motor neurons, pedal serotonergic neurons involved in the peripheral modulation of swimming, and to the serotonergic heart excitor neuron. 3. Inhibitory and excitatory effects appear to be produced by serotonin because they are mimicked by exogenous serotonin and are blocked by the serotonin antagonist mianserin. 4. All serotonergic neurons identified thus far in the CNS of Clione appear to function in a coordinated manner, altering a variety of neural centers all directed toward the activation of swimming behavior. PMID- 8714634 TI - Quantitative effects of GABA and bicuculline methiodide on receptive field properties of neurons in real and simulated whisker barrels. AB - 1. Carbon fiber multibarrel glass microelectrodes were used to record extracellular single-unit activity during microiontophoretic application of gamma aminobutyric acid (GABA) or bicuculline methiodide (BMI) onto layer IV barrel neurons in the somatosensory cortex of fentanyl-sedated rats. Excitatory and inhibitory aspects of the neurons' receptive fields were quantified with the use of controlled whisker stimuli. The principally activating whisker and one of its immediately adjacent neighbors were deflected alone or in paired combinations involving a condition-test paradigm. 2. Units were distinguished electrophysiologically on the basis of the time course of their action potential waveforms. Data were obtained from 26 regular-spike units (RSUs; presumed spiny stellate cells) and 7 fast-spike units (FSUs; presumed GABAergic neurons). An average of 15.0 nA of GABA produced a one-third to one-half reduction in RSU responses evoked by the maximally effective stimulus. An average of 8.7 nA of BMI was needed to counteract this reduction. This amount of BMI, in the absence of exogenous GABA, was found to increase average RSU and FSU responses by 98 and 53%, respectively, relative to predrug levels. 3. For RSUs, the BMI-induced twofold increase in responses evoked by moving the principal whisker at the neuron's best deflection angle was accompanied by an almost threefold increase in responses evoked by similarly moving an adjacent whisker. Disproportionately large percentage increases were also seen for responses to nonpreferred directions of principal and adjacent whisker movement. BMI thus effectively increased receptive field size and decreased angular tuning. Similarly, responses to stimulus offsets, which are normally smaller than ON responses, were increased proportionally more. 4. Predrug responses of FSUs were more vigorous than those of RSUs. However, FSUs showed a similar inverse relationship between percentage increase with BMI and initial response magnitude, although the proportional increases were less pronounced. 5. GABA, like BMI, had the greatest proportional effects on those responses that were initially smallest. It produced results opposite those of BMI, effectively decreasing receptive field size and sharpening angular tuning. 6. A previously described computational model of a barrel was tested for its ability to reproduce quantitatively the effects of BMI and GABA. The application of BMI was simulated by decreasing the strength of the inhibitory inputs onto the particular cell under study in the model network. GABA microiontophoresis was simulated by adding a constant hyperpolarizing voltage. The model RSUs and FSUs displayed proportional changes in response magnitude that were quantitatively similar to those of their biological counterparts. 7. Surround inhibition was greatly attenuated by BMI application, both for the real and simulated barrel neurons. Disinhibition was less pronounced for the former, perhaps because, unlike the simulated neurons, they also possess GABAB receptors, which are unaffected by BMI. 8. We conclude that the inhibitory receptive field properties of barrel neurons can be explained by intrabarrel inhibition and that the expansion of receptive field size and loss of angular tuning with BMI is due to an enhanced effectiveness of convergent, multi-whisker thalamocortical input. Examination of the model neurons' behavior suggests that the altered activity in response to GABA or BMI application, respectively, can be explained by the nonlinear effects of shifting somal membrane potential away from or toward the neuron's firing threshold. PMID- 8714635 TI - Plasticity in the multifunctional buccal central pattern generator of Helisoma illuminated by the identification of phase 3 interneurons. AB - 1. The mechanism for generating diverse patterns of buccal motor neuron activity was explored in the multifunctional central pattern generator (CPG) of Helisoma. The standard pattern of motor neuron activity, which results in typical feeding behavior, consists of three distinct phases of buccal motor neuron activity. We have previously identified CPG interneurons that control the motor neuron activity during phases 1 and 2 of the standard pattern. Here we identify a pair of interneurons responsible for buccal motor neuron activity during phase 3, and examine the variability in the interactions between this third subunit and other subunits of the CPG. 2. During the production of the standard pattern, phase 3 excitation in many buccal motor neurons follows a prominent phase 2 inhibitory postsynaptic potential. Therefore phase 3 excitation was previously attributed to postinhibitory rebound (PIR) in these motor neurons. Two classes of observations indicated that PIR was insufficient to account for phase 3 activity, necessitating phase 3 interneurons. 1) A subset of identified buccal neurons is inhibited during phase 3 by discrete synaptic input. 2) Other identified buccal neurons display discrete excitation during both phases 2 and 3. 3. A bilaterally symmetrical pair of CPG interneurons, named N3a, was identified and characterized as the source of phase 3 postsynaptic potentials in motor neurons. During phase 3 of the standard motor pattern, interneuron N3a generated bursts of action potentials. Stimulation of N3a, in quiescent preparations, evoked a depolarization in motor neurons that are excited during phase 3 and a hyperpolarization in motor neurons that are inhibited during phase 3. Hyperpolarization of N3a during patterned motor activity eliminated both phase 3 excitation and inhibition. Physiological and morphological characterization of interneuron N3a is provided to invite comparisons with possible homologues in other gastropod feeding CPGs. 4. These data support a model proposed for the organization of the tripartite buccal CPG. According to the model, each of the three phases of buccal motor neuron activity is controlled by discrete subsets of pattern-generating interneurons called subunit 1 (S1), subunit 2 (S2), and subunit 3 (S3). The standard pattern of buccal motor neuron activity underlying feeding is mediated by an S1-S2-S3 sequence of CPG subunit activity. However, a number of "nonstandard" patterns of buccal motor activity were observed. In particular, S2 and S3 activity can occur independently or be linked sequentially in rhythmic patterns other than the standard feeding pattern. Simultaneous recordings of S3 interneuron N3a with effector neurons indicated that N3a can account for phase-3-like postsynaptic potentials (PSPs) in nonstandard patterns. The variety of patterns of buccal motor neuron activity indicates that each CPG subunit can be active in the absence of, or in concert with, activity in any other subunit. 5. To explore how CPG activity may be regulated to generate a particular motor pattern from the CPG's full repertoire, we applied the neuromodulator serotonin. Serotonin initiated and sustained the production of an S2-S3 pattern of activity, in part by enhancing PIR in S3 interneuron N3a after the termination of phase 2 inhibition. PMID- 8714636 TI - Determinants of semicircular canal afferent response dynamics in the toadfish, Opsanus tau. AB - 1. Present results determine the relative contributions of the biomechanical and the posttransduction-current (PTC) mechanisms to the sensory process carried out by the horizontal semicircular canal (HC) in the oyster toadfish, Opsanus tau. The role of each element was estimated using in vivo measurements of hair cell receptor potentials and afferent responses elicited by electrical stimuli and mechanical HC indentation. Individual afferent response dynamics are defined here using first-harmonic transfer functions presented in the form of response gain and phase for sinusoidal stimuli from approximately 0.02-30 Hz. Comparison of the response dynamics for the two types of stimuli distinguishes the mechanical and the PTC transfer functions leading to the neural response. The results show that both mechanisms contribute significantly to the overall signal processing performed by the semicircular canals. 2. Endolymphatic polarization and HC indentation. Modulation of the endolymphatic potential by current injection induces a differential voltage across the apical face of the hair cells that drives the transduction current directly via the Nernst-Planck potential. Results show that the electrical impedance of the apical tight junctions is much larger than the basal impedance to ground in O. tau, such that leakage current to the basolateral space is negligible and the voltage-sensitive basolateral currents remain fully functional during polarization of the endolymph (in the frequency range tested). Extracellular afferent responses to endolymphatic polarization were combined with responses to HC indentation to separate the relative contributions of the mechanical and the PTC mechanisms to the overall afferent response dynamics. Data show that more than one-half of the overall signal processing, as defined by the first-harmonic transfer function, persists even when canal mechanics is bypassed. 3. Hair-cell receptor potential modulation during HC indentation. Sharp microelectrodes were used to record the modulation of hair-cell receptor potentials (intracellular voltages) in vivo during physiological levels of sinusoidal HC indentation. Receptor potentials exhibit modulations dominated by the first harmonic and centered about the resting potential. The average gain of the receptor-potential modulation for HC indentation is approximately 0.88 mV/microns indent, corresponding to a value of 0.22 mV/deg/s head velocity, centered near zero phase over the range tested from 0.1-10 Hz. The present receptor potential data fall well short of spanning the full range of gain and phase present in the afferent population. Rather, intracellular hair-cell responses are consistent with the frequency-dependent mechanical activation of the transduction current as determined above. 4. Origins of individual afferent responses. The population of afferent responses forms a continuous distribution that is discussed here in terms of three groups as defined by Boyle and Highstein: velocity-sensitive low gain (LG) afferents, velocity/acceleration-sensitive high gain (HG) afferents, and acceleration sensitive (A) afferents. The response dynamics of individual afferents were found to be determined by a mix of biomechanical and biophysical factors that vary systematically between these afferent groups. All afferents show low-frequency phase advancement and gain decrease during HC indentation associated with the mechanical lower-corner frequency and high-frequency phase and gain enhancements associated with the PTC processing. In highly phase-advanced afferents (A type), the mechanical response is additive with the PTC processing to achieve broad-band acceleration sensitive neural responses.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8714637 TI - Monte Carlo simulation of fast excitatory synaptic transmission at a hippocampal synapse. AB - 1. A simulation of fast excitatory synaptic transmission at a hippocampal synapse is presented. Individual neurotransmitter molecules are followed as they diffuse through the synaptic cleft and interact with the postsynaptic receptors. The ability of the model to reproduce published results of patch-clamp experiments on CA3 pyramidal cells is illustrated; parameters of the model that affect the time course and variability of the excitatory postsynaptic current (EPSC) are then investigated. 2. To simulate an EPSC, we release 4,000 neurotransmitter molecules simultaneously from a point source centered 15 nm above a rectangular grid of 14 x 14 postsynaptic receptors. The simulated EPSC at room temperature has a 10-90% rise time of 0.28 ms and a peak open probability of 0.27, and decays with a time constant of 2.33 ms, comparing well with values in the literature. 3. To simulate changes in temperature, we use a 10 degrees temperature coefficient (Q10) for diffusion of 1.3 and apply a Q10 of 3.0 to all the rate constants of the kinetic scheme. At 37 degrees C, the 10-90 rise time is 0.07 ms, the peak open probability is 0.56, and the decay time constant is 0.70 ms. The coefficient of variation (CV) at the peak of the EPSC is 9.4% at room temperature; at 37 degrees C, the CV at the peak drops to 6.6%. 4. We use the diffusion coefficient of glutamine, 7.6 x 10(-6) cm2/s, to model the random movement of glutamate molecules in the synaptic cleft. Slower rates of diffusion increase the peak response and slow the time course of decay of the EPSC. 5. Random variations in release site position have little effect on the time course of the average EPSC or on the CV of the peak response. We simulate a dose-response curve for the effects of releasing between 100 and 7,500 neurotransmitter molecules per vesicle. The half-maximal response occurs for 1,740 molecules. For a simulation with 100 postsynaptic receptors and a diffusion coefficient of 2.0 x 10(-6) cm2/s, 4,000 molecules approaches a saturating dose. 6. Changes to the width of the synaptic cleft, or to the number and spacing of the postsynaptic receptors, have marked effects on the peak height of the simulated EPSC. 7. We extend the model to include a spherical vesicle (50 nm diam) connected to the synaptic cleft by a cylindrical pore 15 nm long. Neurotransmitter molecules are randomly distributed within the vesicle and allowed to diffuse into the synaptic cleft through the pore, which opens to its full diameter in one time step. We find that the pore must open to a diameter of > or = 7 nm within 1 microsecond in order to match the time courses of EPSCs in the literature. PMID- 8714638 TI - Whole cell and single-channel properties of a unique voltage-activated sustained calcium current identified in teleost retinal horizontal cells. AB - 1. A voltage-activated, sustained calcium current in white bass retinal cone horizontal cells was characterized on the basis of electrophysiological and pharmacological criteria. Studies were performed with the use of a combination of whole cell and single-channel analysis of outside-out excised patches from isolated, cultured retinal horizontal cells. 2. We found that the white bass sustained calcium channel represents a unique type of calcium channel. On the basis of our analysis, it does not fall into any current classification scheme. The horizontal cell channel shares some biophysical and pharmacological properties with the typical high-voltage-activated L-type channel, but it also has features in common with the P-type channel. 3. The biophysical characteristics of the channel were most typical of an L-type channel. It activated above -30 mV membrane potential and only very slowly inactivated. It had a single-channel conductance of 25 pS. 4. Like the typical L-type current, the horizontal cell current was sensitive to the dihydropyridine agonist Bay K 8644. It prolonged the channel open time, which resulted in a large increase in macroscopic current flow into the cell. However, unlike the typical L current, dihydropyridine antagonists (nifedipine, nimodipine, etc.) as well as the specific L-channel inhibitor diltiazem were only moderately effective at best. 5. In a previous study, we found the current was antagonized by a factor found in funnel-web spider toxin. Here we show that the current is completely blocked by low doses of omega-agatoxin IVA. These are characteristics of the P-type calcium channel. But unlike the P current, the horizontal cell current is relatively insensitive to low or high doses of omega-conotoxin MVIIC. 6. The overall combination of calcium channel characteristics sets apart the calcium channel in bass horizontal cells from previously described channels. It appears to be a unique, tissue-specific ion channel, which we have labeled the PL channel. PMID- 8714639 TI - Restoration of extensor excitability in the acute spinal cat by the 5-HT2 agonist DOI. AB - 1. The decerebrate cat preparation with an intact spinal cord is characterized by a high degree of excitability in extensor motoneuron pools, which is eliminated by acute spinalization. Subtype-specific agonists for serotonin (5-HT) were investigated in terms of their effectiveness in restoring the extensor excitability following spinalization. 2. Our hypothesis was that 5-HT2 receptors have the primary role in enhancement of extensor reflex excitability, whereas 5 HT1A and 5-HT1B/D receptors are relatively unimportant. Reflex excitability was assessed from the tonic levels of force and electromyographic (EMG) output from the ankle extensors medial gastrocnemius (MG) and soleus (SOL), and from the reflex forces in both these muscles generated by ramp-and-hold stretches of MG. 3. Before spinal transection, MG and SOL usually exhibited a small amount of tonic background EMG activity and force output. Ramp-and-hold stretch of MG generated a large-amplitude reflex response. Spinal transection at the level of T10 virtually abolished tonic background activity in both extensors and greatly attenuated the MG stretch reflex. Ventral topical application of the selective 5 HT2A/2C agonist (+-)-1-(2,5-dimethoxy-4-iodophenyl)-2-amino-propane hydrochloride (DOI) restored the amplitude of the MG stretch reflex in a dose-dependent fashion. However, a considerable portion of the DOI-mediated restoration of MG stretch reflex force was due to elevation of tonic background force levels above previous intact cord levels. 4. The DOI-induced increase in extensor tonic background excitability and facilitation of MG stretch reflex were reversed by ventral topical administration of the selective 5-HT2 antagonist ketanserin. No increase in extensor excitability was observed in spinalized preparations after administration of either the 5-HT1A agonist (+-)-8-hydroxy-dipropylaminotetralin hydrobromide or the 5-HT1B/1D agonist 7-trifluoromethyl-4-(4 methyl-1 piperazinyl)-pyrrolo[1,2- a]quinoxaline maleate. These data strongly suggest that the DOI-induced facilitation of extensor stretch reflex and tonic activity in spinalized preparations is mediated through an action on spinal 5-HT2 receptors. 5. One important difference between the actions of DOI in spinalized versus intact states was that the DOI-induced tonic and reflex forces in the spinalized state were subject to irregular oscillations. In contrast, DOI did not noticeably affect the smoothness of reflex force generation in the intact state. This discrepancy was probably due to the effects of clasp knife inhibition from muscular free nerve endings, which have potent reflex actions in the spinalized but not intact states. Thus DOI elevated excitability levels but did not alter the effects of spinalization on stretch reflex patterns. PMID- 8714640 TI - Activation of NMDA receptors produces dopamine-mediated changes in fish retinal horizontal cell light responses. AB - 1. The action of N-methyl-D-aspartate (NMDA) on cone horizontal cells was studied in in vitro goldfish retinas superfused with a bicarbonate-based Ringer solution that contained D-serine (0.5 microM), a glycine analogue, but no added Mg2+. Horizontal cell light responses and electrical coupling were assessed by monitoring responses to full-field stimuli and to small, centered (0.4 mm diam) spots of light, respectively. 2. NMDA uncoupled horizontal cells, reduced their light responsiveness, and acted in a dose-dependent manner, with threshold at 10 microM and maximum effect at 100 microM. 3. Application of the NMDA antagonists DL-2-amino-7-phosphonoheptanoic acid-AP-5) or D-2-amino-5-phosphonopentanoic acid AP-5) (50 microM) blocked the uncoupling action of NMDA (100 microM), as did prior application of SCH23390, a dopamine D1 antagonist, or prior treatment of the retinas with 6-hydroxydopamine, a procedure that destroys dopaminergic neurons. 4. Addition of Mg2+ (1 mM) partially blocked the effects of NMDA at 100 microM and completely blocked the effects of 50 microM NMDA. The effects of NMDA (50 or 100 microM) were also reduced if it was applied without D-serine. 5. Both flickering (5 Hz) and sustained light stimulation uncoupled horizontal cells and reduced their light responsiveness. Application of AP-7 blocked the effects of flickering light stimulation, but did not block the effects of sustained light. 6. These results suggest that activation of NMDA receptors in the fish retina uncouples cone horizontal cells and decreases their light responsiveness by increasing dopamine release. The results further suggest that flickering light, but not sustained light, increases the release of dopamine through activation of NMDA receptors. PMID- 8714641 TI - Spontaneous rhythmic bursts induced by pharmacological block of inhibition in lumbar motoneurons of the neonatal rat spinal cord. AB - 1. The effects of blocking gamma-aminobutyric acid- and glycine-mediated synaptic transmission by bicuculline and strychnine on the neonatal rat isolated spinal cord were investigated by intracellular recording from motoneurons with the use of current-clamp and voltage-clamp techniques and by extracellular recording from homologous ventral roots of the L5 segment. 2. Bicuculline per se evoked irregular bursts of motoneuron membrane potential, often comprising individual events fused together. Strychnine alone did not elicit spontaneous bursting in the large majority of preparations. Simultaneous application of bicuculline and strychnine consistently induced regular rhythmic bursts (frequency approximately 2 per min, duration approximately 7 s), comprising a rapid depolarization followed by large-amplitude oscillations. 3. Burst frequency, duration, and intraburst oscillation time course were independent of motoneuron membrane potential. Burst and oscillation amplitude decreased with membrane depolarization and, under voltage-clamp conditions, inverted polarity near 0 mV. 4. The regular bursts produced by bicuculline and strychnine were blocked by 6-cyano-7 nitroquinoxaline-2,3-dione, tetrodotoxin, or Cd2+. 5. N-methyl-D-aspartate antagonists [R-5-aminophosphonovalerate or 3-((RS)-2-carboxypiperazine-4-yl) propyl-1-phosphonate (CPP)] reversibly blocked or slowed down bursting induced by bicuculline and strychnine. Addition of cyclothiazide to the bicuculline and strychnine solution increased bursting frequency while preserving the regular burst structure; under these conditions bursts became insensitive to CPP. 6. In the presence of bicuculline and strychnine, 5-hydroxytryptamine (5-HT) increased burst frequency and decreased burst duration in a dose-dependent fashion. 7. In the presence of bicuculline and strychnine, L5 ventral roots developed synchronous rhythmic activity with a time course similar to that recorded from individual motoneurons. The rhythmic activity was accelerated by 5-HT on both roots, in accordance with observations on single motoneurons. 8. Rhythmic bursts thus appear to result from large, synchronous synaptic events generated by a network modulated by 5-HT and highly sensitive to variations in efficacy of glutamatergic synaptic transmission. These results show that in the rat spinal cord highly patterned motor output can occur despite block of inhibition. PMID- 8714642 TI - Serotonin modulates N- and P-type calcium currents in neocortical pyramidal neurons via a membrane-delimited pathway. AB - 1. The effects of serotonin (5HT) on neocortical pyramidal neurons were studied using whole cell and ON-cell patch-clamp recordings from acutely dissociated neurons. 2. 5HT decreased high voltage-activated calcium channel currents in a dose-dependent and reversible manner in acutely dissociated neocortical pyramidal neurons. The maximum block was 30% of the peak whole cell current (at -10 mV). 3. The 5HT modulation was mimicked by 5HT1A agonists and was reduced by 5HT1A antagonists. 5HT2 antagonists had no effect on the modulation. These data suggest that the 5HT effects were mediated by 5HT1A receptors. 4. The 5HT1A modulation was reduced in the presence of the specific N-type blocker omega-conotoxin GVIA (CgTx) and by the P-type channel blocker omega-agatoxin IVA (AgTx), but not by the L-type blocker nifedipine. 5HT did not modulate the slowed tail currents in the presence of the dihydropyridine agonist Bay K 8644. These data suggest that N and P-type channels (but not L-type channels) were targeted by 5HT. 5. The modulation involved G proteins and utilized a membrane-delimited pathway. The modulation was rapid in onset (tau approximately 600 ms) and offset. About 50% of the reduction in current by 5HT1A agonists was overcome by prepulses to 120 mV. 6. Slowing of current onset kinetics in response to 5HT1A agonists was seen rarely in neocortical pyramidal neurons (11% of cases). The presence of slowing depended on agonist concentration, being evident only with high micromolar doses. PMID- 8714643 TI - Two ion currents activated by acetylcholine in the ARC muscle of Aplysia. AB - 1. This work continues our examination of the electrophysiology and contractions of single fibers dissociated from a widely studied molluscan muscle, the accessory radula closer (ARC) muscle of Aplysia californica, aimed at understanding its excitation-contraction mechanisms and their modulation. 2. Extensive previous work has characterized a number of basal ion currents present in the fibers and effects of transmitters and peptide cotransmitters that modulate ARC-muscle contractions in vivo. Here we use current clamp, voltage clamp, and contraction measurements to examine the actions of acetylcholine (ACh), the transmitter that induces the contractions. 3. As in the whole ARC muscle, ACh depolarizes unclamped fibers maximally to about -25 mV where, no matter how much ACh is applied, the depolarization saturates. 4. The underlying ACh-activated current is in fact the sum of two quite distinct components, IACh,cat and IACh,Cl. 5. IACh,cat is itself a mixed current carried by cations (physiologically mainly by Na+, but to a significant degree also by Ca2+), reverses near +20 mV, rectifies inwardly, exhibits prominent voltage-dependent kinetics of activation with hyperpolarization, and is selectively blocked by hexamethonium. 6. In contrast, IACh,Cl is carried by Cl-, reverses near -60 mV, exhibits little rectification or voltage-dependent kinetics, is activated selectively by suberyldicholine, and is blocked by alpha-bungarotoxin. 7. Both currents activate fast when ACh is applied, desensitize relatively slowly in its presence, then deactivate fast. Both currents are activated at similar ACh concentrations, half-maximally at approximately 10 microM. Both currents also are activated by carbachol and propionylcholine and blocked by d-tubocurarine, bicuculline and paraoxon. Picrotoxin and atropine block IACh,cat better, 4 acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic acid (SITS), 4,4' diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS), and anthracene 9-carboxylic acid IACh,Cl better. 8. The two currents are virtually identical to ACh-activated cationic (Na) and Cl currents that are ubiquitous in molluscan neurons. As has been proposed for the neuronal currents, IACh,cat resembles vertebrate neuronal nicotinic ACh-receptor (nAChR) currents, whereas IACh,Cl resembles vertebrate skeletal muscle nAChR currents. 9. Functionally, we believe that IACh,cat serves primarily to depolarize the ARC muscle to open voltage-activated L-type Ca channels, allow Ca2+ influx, and initiate contraction. Physiologically significant Ca2+ may also enter through the ACh,cat channels themselves. 10. By superimposing on IACh,cat, IACh,Cl brings the reversal potential of the combined current to around -25 mV and thereby sets a relatively negative upper limit to the ACh-induced depolarization. We propose that this is its physiological role. By limiting the depolarization, IACh,Cl limits the degree of activation of the Ca current and Ca2+ influx, and so prevents excessive contraction. More importantly, it moderates the voltage during contraction to a range where small voltage changes can finely grade contraction amplitude in this nonspiking muscle. 11. Indeed, in contraction experiments on the single fibers, there is an inverse correlation between the IACh,Cl/IACh,cat ratio and the magnitude of the ACh induced depolarization and contraction. Furthermore, increased pharmacological activation of IACh,Cl depresses, and block of IACh,Cl enhances, both the depolarization and contraction. 12. Obligatory simultaneous coactivation of IACh,cat and IACh,Cl in the ARC muscle may be part of a peripheral control mechanism that automatically keeps the size of its contractions within behaviorally optimal limits. PMID- 8714644 TI - Adaptive control for backward quadrupedal walking VI. metatarsophalangeal joint dynamics and motor patterns of digit muscles. AB - 1. We compared the dynamics of the metatarsophalangeal (MTP) joint of the cat's hind paw and the motor patterns of two short and four long muscles of the digits for two walking forms, forward (FWD) and backward (BWD). Kinematic (angular displacements) data digitized from high-speed cine film and electromyographic (EMG) data were synchronized and assessed for bouts of treadmill walking. Kinetic data (joint forces) were calculated from kinematic and anthropometric data with the use of inverse-dynamic calculations in which the MTP joint net torque was divided into gravitational, motion-dependent, ground contact (absent for swing), and muscle torque components. Swing-phase kinetics were calculated from treadmill steps and stance-phase kinetics from overground steps in which one hind paw contacted a miniature force platform embedded in the walkway. 2. The plantar angle at the intersection of the metatarsal and phalangeal segmental lines was used to measure MTP angular displacements. During swing for both walking forms, the MTP joint flexed (F) and then extended (E); however, the F-E transition occurred at the onset of FWD swing and at the end of BWD swing. For FWD walking, the MTP joint extended at a constant velocity during most of stance as the cat's weight rotated forward over the paw. During the unweighting phase at the end of stance, the MTP joint flexed rapidly before paw lift off. For BWD walking, the MTP joint extended briefly at stance onset (similar to a yield) and then flexed at a constant velocity as the cat's weight rotated backward over the paw. At the end of stance, the MTP joint extended and then flexed slightly as the paw was unweighted before paw lift off. 3. For both forms of walking, three of the six muscles tested were recruited just before paw contact and remained active for most (75-80%) of stance for both walking forms: plantaris (PLT), flexor hallucis longus (FHL), and flexor digitorum brevis (FDB). Their recruitment contributed to the flexor muscle torque at the MTP joint during most of FWD and BWD stance and was responsible for the absorption of mechanical power at the MTP joint for FWD stance and generation of mechanical power at the MTP joint during BWD stance. Also, the FHL and PLT, along with the soleus (SOL; also recorded in this study), contributed to an extensor muscle torque (described in paper IV of this series) and the generation of mechanical power at the ankle joint during stance of FWD and BWD walking. 4. The timing of activity for three muscles recruited during FWD swing was distinct for the two walking forms. The hallmark burst of the flexor digitorum longus (FDL)--a single burst, brief in duration and high in amplitude- occurred at the end of FWD swing (as the toes flexed rapidly) but shifted to the onset of BWD stance (as the claws protruded and toes extended) during paw weighting. The extensor digitorum longus (EDL) was recruited after paw off and was active for most of FWD swing; its activity contributed to an extensor muscle torque at the MTP joint and a flexor muscle torque at the ankle joint. For BWD walking, EDL recruitment shifted to an earlier phase in the step cycle and coincided with toe extension, which occurred at the end of stance before paw lift off. This pre-lift off activity continued into the first part of swing and contributed to an extensor muscle torque at the MTP joint and a flexor muscle torque at the ankle.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8714645 TI - Role of the afterhyperpolarization in control of discharge properties of septal cholinergic neurons in vitro. AB - 1. The properties of the cholinergic neurons of the rat medial septum and nucleus of the diagonal band of Broca (MS/DBB) were studied using whole cell patch-clamp recordings in an in vitro slice preparation. 2. Both the transmitter phenotype and the intrinsic membrane properties of 56 MS/DBB neurons were determined post hoc by visualizing intracellularly deposited biocytin with fluorescent avidin and endogenous choline acetyltransferase with immunofluorescence. Twenty seven of 28 MS/DBB neurons exhibiting both a prominent slow afterhyperpolarization (sAHP) following a single action potential and anomalous rectification were identified as cholinergic. The remaining 28 neurons exhibited other intrinsic membrane properties and none were choline acetyltransferase immunoreactive. 3. The sAHP in MS/DBB cholinergic neurons was blocked reversibly either by reducing extracellular calcium or addition of 100 microM cadmium and irreversibly blocked by 30 nM apamin, suggesting that the sAHP is produced by an apamin-sensitive calcium-activated potassium conductance. 4. MS/DBB cholinergic neurons also exhibited a postspike depolarizing afterpotential (DAP) preceeding the sAHP. Both the DAP and the sAHP were blocked when extracellular calcium was lowered as well as in the presence of 10-50 microM NiCl2. Application of 500 nM omega-conotoxin also reduced the sAHP, while leaving the DAP intact. These data suggest that both transient and high-threshold calcium conductances contribute to generation of the sAHP. 5. When depolarized, cholinergic neurons fired slowly (2-4 Hz) and regularly with little evidence of spike frequency adaptation. When the sAHP was blocked with apamin, the instantaneous frequency of firing increased and the neuron now exhibited prominent spike frequency adaptation. 6. Serotonin (5-HT) reversibly suppressed the sAHP in MS/DBB cholinergic neurons and altered the firing pattern from slow regular discharge to one which exhibited modest spike frequency adaptation. 7. It was concluded that the sAHP limits the firing rate of MS/DBB cholinergic neurons and that physiologically relevant supression of the sAHP by 5-HT may result in state-dependent changes in the discharge pattern of MS/DBB cholinergic neurons. PMID- 8714646 TI - Histamine depolarizes cholinergic septal neurons. AB - 1. Bath application of 10 microM histamine (HA) resulted in a depolarization or inward current in 58/59 cholinergic neurons located in the medial septum and nucleus of the diagonal band of Broca (MS/DBB) in a slice preparation of rat brain. 2. In bridge mode, the histamine-induced depolarization consisted of both fast and slow phases; inward currents that followed the comparable time course were observed under voltage-clamp conditions. The fast depolarization was associated with variable changes in input resistance, while the slow depolarization always was associated with an increase in input resistance. 3. Both fast and slow responses persisted in the presence of tetrodotoxin (TTX), but only the fast response persisted when transmitter release was abolished by bathing the slice in either a low-Ca(2+)-, high-Mg(2+)-containing medium or one containing Cd2+. 4. When ramp voltage-clamp commands were applied during the fast depolarization, the resultant current-voltage (I-V) curves did not intersect over the range of membrane potentials from -130 to -30 mV. Ionic substitution experiments suggested that the bulk of the ionic current flowing during the fast depolarization was carried by sodium ions. 5. The I-V characteristics of the slow inward current identified it as a reduction in an inwardly rectifying potassium conductance. 6. The fast depolarization was significantly reduced by the H1 receptor antagonists pyrilamine and promethazine, but not by the H2 receptor antagonist cimetidine. Neither the H2 receptor agonist impromidine nor the H3 receptor agonist R-alpha-methylhistamine mimicked the response to HA. None of the agonists or antagonists had any observable effect upon the slow depolarization. 7. We conclude that HA directly depolarizes cholinergic MS/DBB neurons by acting as an H1 receptor, which primarily couples to an increase in a TTX-insensitive Na+ conductance. Additionally, HA evokes a slow depolarization mediated by a decrease in an inwardly rectifying potassium conductance but is not generated by activation of classically defined HA receptor subtypes. PMID- 8714647 TI - Role of sympathetic postganglionic neurons in synovial plasma extravasation induced by bradykinin. AB - 1. Plasma extravasation through the endothelium of blood vessels is an integral component of the inflammatory response and is dependent to a large extent on the inflammatory mediator bradykinin (BK). We studied plasma extravasation induced by BK perfusion (BK-induced PE) of the rat knee joint before and after various interventions that affect the sympathetic supply to the knee joint. We tested the hypothesis that plasma extravasation is dependent on the local sympathetic terminal supply to the synovia but not directly dependent on more proximal activity in the sympathetic neurons. As a control we used plasma extravasation induced by platelet activating factor (PAF), which acts directly on the endothelium of the blood vessels, that is, its action is independent of any innervation. Plasma extravasation into the knee joint cavity was determined spectrophotometrically by measuring, over time, the concentration of Evans blue dye extravasation into the joint perfusate following intravenous injection of the dye. 2. Surgical sympathectomy at the lumbar level (L2-L4), performed 4 and 14 days previously, reduced BK-induced PE by approximately 55-70%. 3. Decentralization of the lumbar sympathetic chain (cutting the preganglionic axons that innervate the postganglionic neurons to the hindlimb), interruption of the lumbar sympathetic chain during infusion of BK, or coperfusion of tetrodotoxin into the knee joint cavity did not reduce BK-induced PE. All three interventions abolish the activity in the sympathetic neurons but leave the peripheral postganglionic terminals in the joint capsule intact. 4. Surgical sympathectomy and decentralization did not affect plasma extravasation induced by the intra articular perfusion with PAF. 5. Electrical stimulation of the lumbar sympathetic chain at frequencies of 0.25-5 Hz, which probably also significantly decreases blood flow through the joint capsule, reduced basal plasma extravasation, BK induced PE and PAF-induced PE. This reduction was frequency dependent and was almost maximal at a stimulation frequency of 1 Hz. 6. In conclusion, BK-induced PE into the rat knee joint is dependent on the presence of intact sympathetic postganglionic nerve terminals innervating the joint capsule and not directly dependent on excitation of these neurons. However, electrical stimulation of the sympathetic neurons reduces the level of plasma extravasation, presumably because of vasoconstriction and decrease of blood flow through the joint capsule. These results indicate that peripheral action of inflammatory mediators on terminals of sympathetic neurons produces a facilitative effect on vascular permeability, whereas centrally generated excitation of these neurons, which depresses blood flow (vasoconstrictor function), decreases plasma extravasation. The effect on blood flow is presumed to occur at the precapillary resistance vessels by vesicular release of transmitter(s). The facilitative effect on permeability occurs at the venules and includes inflammatory-mediator-stimulated, non vesicular-dependent production and release of a chemical substance (probably prostaglandin E2). Whether both functions are represented in the same class of sympathetic postganglionic neuron or in distinct ones remains to be elucidated. PMID- 8714648 TI - Anatomy and physiology of the primate interstitial nucleus of Cajal I. efferent projections. AB - 1. The efferent projections of the interstitial nucleus of Cajal (NIC) were studied in the squirrel monkey after iontophoretic injections of biocytin and Phaseolus Vulgaris leucoagglutinin into the NIC. To ensure the proper placement of the tracer, the same pipettes were used to extracellularly record the discharge pattern of NIC neurons. 2. Three projection systems of the NIC were distinguished: commissural (through the posterior commissure), descending, and ascending. 3. The posterior commissure system gave rise to dense terminal fields in the contralateral NIC, the oculomotor nucleus, and the trochlear nucleus. 4. The descending system of NIC projections deployed dense terminal fields in the ipsilateral gigantocellular reticular formation and the paramedian reticular formation of the pons, as well as in the ventromedial and commissural nuclei of the first two spinal cervical segments. It also gave rise to moderate or weak terminal fields in the vestibular complex, the nucleus prepositus hypoglossi, the inferior olive, and the magnocellular reticular formation, as well as cell groups scattered along the paramedian tracts in the pons and the pontine and medullary raphe. 5. The ascending system of NIC projections gave rise to dense terminal fields in the ipsilateral mesencephalic reticular formation and the zona incerta as well as moderate or weak terminal fields in the ipsilateral centromedian and parafascicular thalamic nuclei. It also provided dense bilateral labeling of the rostral interstitial nucleus of the medial longitudinal fasciculus and the fields of Forel, and moderate or weak bilateral labeling of the mediodorsal, central medial, and central lateral nuclei of the thalamus. 6. Models of saccade generation that rely on feedback from the velocity-to-position integrators and include the superior colliculus in their local feedback loop are contradicted because no fibers originating from the NIC traveled to the superior colliculus to deploy terminal fields. 7. Consistent with its morphological and functional diversity, these data indicate that the primate NIC sends signals to a multitude of targets implicated in the control of eye and head movements. PMID- 8714649 TI - Effects of glucose deprivation on NMDA-induced current and intracellular Ca2+ in rat substantia nigra neurons. AB - 1. The effects of glucose deprivation on N-methyl-D-asparate (NMDA)-induced current (INMDA) and the intracellular free Ca2+ concentration ([Ca2+]i) in the acutely dissociated rat substantia nigra neurons were investigated using the nystatin-perforated patch-clamp technique under voltage clamp and the microfluometry with a fluorescent probe, Indo-1. 2. Application of NMDA induced a peak and a successive steady-state inward current, and an outward current immediately after washout at a holding potential of -40 mV. The amplitudes of the three current components of INMDA were increased by increasing the concentrations of NMDA with half-maximum concentrations (EC50s) of 1.1 x 10(-4) M, 1.2 x 10(-4) M, and 1.6 x 10(-4) M, respectively. 3. The reversal potentials of the peak inward and outward currents were -4 +/- 3 (SE) mV and -76 +/- 2 mV, respectively. The latter was close to the theoretical K+ equilibrium potential (-82 mV). 4. The outward current was potentiated by increase in extracellular Ca2+ concentration and was blocked by Cs+ internal solution and suppressed by 5 x 10(-3) M tetraethylammonium chloride and 10(-7) M charybdotoxin, indicating that it was Ca(2+)-activated K+ current. 5. Application of NMDA increased [Ca2+]i in a concentration-dependent manner with an EC50 of 3.9 x 10(-5) M. 6. Depriving the external solution of glucose induced a slowly developing outward current and increased the basal level of [Ca2+]i. It also prolonged the NMDA-induced outward current without affecting the peak inward current, and prolonged the NMDA-induced increase in [Ca2+]i without changing the peak [Ca2+]i. 7. These findings suggest that the deprivation of glucose did not affect the NMDA-induced influx of Ca2+ into the cells, but it inhibited Ca2+ clearance by affecting the efflux of Ca2+ to the extracellular space, reuptake into the intracellular Ca2+ stores, and/or active extrusion from intracellular stores. PMID- 8714650 TI - Propagation of spindle waves in a thalamic slice model. AB - 1. We study the propagation and dynamics of spindle waves in thalamic slices by developing and analyzing a model of reciprocally coupled populations of excitatory thalamocortical (TC) neurons and inhibitory thalamic reticular (RE) neurons. 2. Each TC neuron has three intrinsic ionic currents: a low-threshold T type Ca+2 current (ICa-T), a hyperpolarization-activated cation ("sag") current (Ih) and a leak current. Each RE cell also has three currents: ICa-T, a leak current, and a calcium-activated potassium current (IAHP). Isolated TC cells are at rest, can burst when released or depolarized from a hyperpolarized level, and burst rhythmically under moderate constant hyperpolarizing current. Isolated RE cells are at a hyperpolarized resting membrane potential and can burst when depolarized. 3. TC cells excite RE cells with fast alpha-amino-3-hydroxy-5-methyl 4-isoxazolepropionic acid (AMPA) synapses, and RE cells inhibit TC cells with fast gamma-aminobutyric acid-A (GABAA) and slow GABAB synapses and inhibit each other with GABAA synapses only. GABAB postsynaptic conductances operate far from saturation, and the slow inhibitory postsynaptic potentials (IPSPs) increase with the width of the presynaptic burst. The model network is a one-dimensional cellular array with localized coupling. The synaptic coupling strength decays with the distance between the pre- and postsynaptic cells, either exponentially or as a step function. 4. The "intact" network can oscillate with partial synchrony and a population frequency of approximately 10 Hz. RE cells emit bursts almost at every oscillation cycle, whereas TC cells do so almost at every other cycle. Block of GABAB receptors hardly changes the network behavior. Block of GABAA receptors leads the network to a slowed oscillatory state, where the population frequency is approximately 4 Hz and both RE and TC cells fire unusually long bursts at every cycle and in full synchrony. These results are consistent with the experimental observations of von Krosigk, Bal, and McCormick. We obtain such consistency only when the above assumptions regarding the synaptic dynamics, particularly nonsaturating GABAB synapses, are fulfilled. 5. The slice model has a stable rest state with no neural activity. By initially depolarizing a few neurons at one end of the slice while all the other cells are at rest, a recruitment process may be initiated, and a wavefront of oscillatory activity propagates across the slice. Ahead of the wavefront, neurons are quiescent; neurons behind it oscillate. We find that the wave progresses forward in a lurching manner. TC cells that have just become inhibited must be hyperpolarized for a long enough time before they can fire rebound bursts and recruit RE cells. This step limits the wavefront velocity and may involve a substantial part of the cycle when no cells at the front are depolarized. 6. The wavefront velocity increases linearly with the characteristic spatial length of the connectivity (the footprint length). It increases only gradually with the synaptic strength, logarithmically in the case of an exponential connection function and only slightly for a step connection function. It also decreases gradually with a potassium leak conductance that hyperpolarizes RE cells. 7. To reproduce the experimentally measured wavefront velocity of approximately 1 mm/s, together with other in vitro observations, both the RE-to-TC and the TC-to-RE projections in the model should be spatially localized. The sum of the RE-to-TC and the TC-to-RE synaptic footprint lengths should be on the order of 100 microns. (ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8714651 TI - Intracellular injection of a Ca2+ chelator prevents generation of anoxic LTP. AB - 1. The effects of intracellular injection of Ca2+ chelator 1,2-bis (2 aminophenoxy) ethane N,N,N',N'-tetra-acetic acid (BAPTA, 50 mM) on anoxia aglycemia-induced long-term potentiation (LTP) were investigated in the CA1 region of hippocampal slices with the use of extra- and intracellular recording techniques. Experiments were performed in artificial cerebrospinal fluid (ACSF) containing 10 microM bicuculline and 10 microM 6-cyano-7-nitroquinoxaline- 2,3 dione (CNQX) to pharmacologically isolate N-methyl-D-aspartate (NMDA)-receptor mediated responses. NMDA-receptor-mediated excitatory postsynaptic potentials (EPSPs) and field potentials were evoked by stimulation of the Schaffer collateral/commissural pathway in the presence of 0.3 mM MgCl2 and 10 microM glycine to promote NMDA-receptor-mediated responses. Under these conditions, application of 50 microM D-2-amino-phosphono-valerate (D-APV) abolished EPSPs and field potentials. 2. Anoxic-aglycemic (AA) episodes (duration 2-2.5 min) potentiated the initial slope (measured within 3 ms from the onset of the synaptic responses) of EPSPs by 108 +/- 14.3% (mean +/- SE, P = 0.0012, n = 7). We refer to this LTP of NMDA-receptor-mediated synaptic responses as anoxic LTP. 3. Intracellular injection of the Ca2+ chelator BAPTA (with the intracellular recording electrode filled with 50 mM BAPTA in 3 M KCl) prevented anoxic LTP. Thirty to 40 min after the AA episode, in BAPTA-loaded cells, the initial slope of the EPSPs was not significantly changed (+7.12 +/- 5%, P = 0.35, n = 5). In contrast, the initial slope of the field potentials, measured at the same time in the same slices, was persistently increased (+49 +/- 2.8%, P = 0.0022, n = 5). 4. High-frequency tetanic stimulation (100 Hz for 500 ms, 2 times, 30 s apart) of the Schaffer collateral/commissural pathway, applied > 0.5 h after the AA episode, induced an additional significant and persistent increase in the initial slope of the field potential (tetanic LTP, +35.4 +/- 9.8%, P = 0.012, n = 5). In BAPTA-loaded cells, there was no further change in the initial slope of the EPSP (+3.9 +/- 3.4%, P = 0.205, n = 5) after the tetanic stimulation. 5. We also report that AA episodes or tetanic stimulation induced a persistent increase in a late synaptic component that was blocked by 50 microM D-APV. This late component was mediated polysynaptically, because its time to peak decreased with increasing stimulation intensities and it was strongly reduced by high-divalent-cation superfusate (ACSF containing 7 mM Ca2+). This component, which had a delay of approximately 8-30 ms, contaminated mainly the peak amplitude and the decay of the monosynaptic response without affecting its initial slope. Thus the measure of the initial slope takes into account only the early phase of the monosynaptic response. 6. We conclude that 1) a rise in intracellular Ca2+ is necessary to generate anoxic LTP of NMDA-receptor-mediated responses, as is the case for tetanic LTP; and 2) in the presence of bicuculline and low extracellular Mg2+, AA episodes and tetanic stimulations induced a long-lasting enhancement of a polysynaptic component mediated or controlled by NMDA receptors. PMID- 8714652 TI - Responses of ventral cochlear nucleus onset and chopper units as a function of signal bandwidth. AB - 1. The responses of units in the ventral cochlear nucleus in anesthetized guinea pigs have been measured to best-frequency tones, noise bands geometrically centered around the unit best frequency, and noise bands asymmetrically positioned around the best frequency. 2. Each unit isolated was characterized using peristimulus time histograms (PSTHs) to best-frequency tones at 20 and 50 dB suprathreshold, frequency-intensity response areas and rate-versus-level functions in response to best-frequency tones and wideband noise. The data reported here are derived from full analyses of 5 chopper units and 17 onset units. The onsets were divided into onset-I (OnI), onset-L (OnL), and onset-C (OnC) by the criteria described by Winter and Palmer: the PSTHs of OnI units show only an onset response, OnL units respond with a single spike at onset followed by a low level of sustained activity, and OnC units have PSTHs with one to four onset peaks and low levels of sustained discharge. 3. In response to geometrically centered noise bands of constant spectral density, the discharge of chopper units and one OnI unit increased over a relatively narrow range of bandwidths, corresponding to the equivalent rectangular bandwidth calculated from their response area, and then became constant. In contrast, OnL and OnC units showed increases in discharge rate with noise bandwidth over very wide ranges of bandwidth. The growth of the discharge rate with noise bandwidth was approximately linear on double logarithmic axes and therefore could be described by a power function with an exponent of 0.37. This relation held even for noise levels near threshold. 4. When noise bands with constant spectral density (at the input to the earphone) were presented with one edge fixed at the unit's best frequency, the discharge rate of most chopper units and the one OnI unit increased over a narrow range of bandwidths and then became constant. This pattern was observed irrespective of whether the second edge of the noise was progressively increased above, or decreased below, the best frequency. For two of the chopper units, in which lateral inhibitory sidebands could be demonstrated, increasing the noise bandwidth led first to increases and then to decreases in the discharge rate as the noise energy impinged upon the sideband. The chopper units act like energy detectors with a filter corresponding to their single tone response area, but, for some units, with the addition of inhibitory sidebands. 5. For the OnL and OnC units, increasing the noise bandwidth above or below best frequency caused progressive increases in the discharge rate over wide ranges of bandwidth. These increases occurred even for low noise spectral densities. The growth in discharge rate for these onset units was well fitted at all spectral density levels by power functions: one above best frequency and one below. At levels of the noise 40 dB above the unit threshold, the point at which the discharge rate reached 90% of its maximum was, on average, about 2 octaves below best frequency and 1 octave above. For some onset units, changes in the discharge rate were seen as the noise bandwidth was varied over about 14 kHz, which is about one-third of the total frequency hearing range of the guinea pig. 6. The data for onset units is consistent with the hypothesis that onset units in the ventral cochlear nucleus achieve their precision in the temporal domain by integration of the inputs from auditory nerve fibers with a wide range of best frequencies. The range of frequency over which onset units integrate frequency matches that of the inhibitory input to dorsal cochlear nucleus neurons, suggesting a possible role as an inhibitory interneuron. PMID- 8714653 TI - Electroresponsive properties and membrane potential trajectories of three types of inspiratory neurons in the newborn mouse brain stem in vitro. AB - 1. The electrophysiological properties of inspiratory neurons were studied in a rhythmically active thick-slice preparation of the newborn mouse brain stem maintained in vitro. Whole cell patch recordings were performed from 60 inspiratory neurons within the rostral ventrolateral part of the slice with the aim of extending the classification of inspiratory neurons to include analysis of active membrane properties. 2. The slice generated a regular rhythmic motor output recorded as burst of action potentials on a XII nerve root with a peak to peak time of 11.5 +/- 3.4 s and a duration of 483 +/- 54 ms (means +/- SD, n = 50). Based on the electroresponsive properties and membrane potential trajectories throughout the respiratory cycle, three types of inspiratory neurons could be distinguished. 3. Type-1 neurons were spiking in the interval between the inspiratory potentials (n = 9) or silent with a resting membrane potential of -48.6 +/- 10.1 mV and an input resistance of 306 +/- 130 M omega (n = 15). The spike activity between the inspiratory potentials was burst-like with spikes riding on top of an underlying depolarization (n = 11) or regular with no evidence of bursting (n = 12). Hyperpolarization of the neurons below threshold for spike initiation did not reveal any underlying phasic synaptic activity, that could explain the bursting behavior. 4. Type-1 neurons showed delayed excitation after hyperpolarizing square current pulses or when the neurons were depolarized from a hyperpolarized level. This membrane behavior resembles the response seen in other CNS neurons expressing an IA. The response to 1-s long depolarizing pulses with a large current strength showed signs of activation of an active depolarizing membrane response leading to a transient reduction in the spike amplitude. The relationship between the membrane potential and the amplitude of square current pulses (Vm-I) showed a small upward rectification below -70 mV, and spike adaptation throughout a 1-s pulse had a largely linear time course. 5. Type-1 neurons depolarized and started to fire spikes 398 +/- 102 ms (n = 20) before the upstroke of the integrated XII nerve discharge. The inspiratory potential was followed by fast hyperpolarization, a short fast-repolarizing phase (1,040 +/- 102 ms, n = 5) and a longer slow-repolarizing phase (lasting until the next inspiratory discharge). 6. Type-2 neurons were spiking in the interval between the inspiratory potentials with no evidence of bursting behavior and had an input resistance of 296 +/- 212 M omega (n = 26). The response to hyperpolarizing pulses revealed an initial sag and postinhibitory rebound depolarization. This membrane behavior resembles the response seen in other CNS neurons expressing an Ih. The Vm-I relationship was linear at depolarized potentials and showed a marked upward rectification below -60 mV. Spike trains elicited by 1-s long pulses showed a pronounced early and late adaptation. 7. Type-2 neurons depolarized and started to fire spikes 171 +/- 87 ms (n = 23) before the upstroke of the integrated XII nerve discharge. The inspiratory potential had a variable amplitude from cell to cell and was followed by a short hyperpolarization in the cells displaying a large amplitude inspiratory potential.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8714654 TI - Thyrotropin-releasing hormone (TRH) depolarizes a subset of inspiratory neurons in the newborn mouse brain stem in vitro. AB - 1. To extend the classification of respiratory neurons based on active membrane properties and discharge patterns to include responses to respiratory modulators, we have studied the effect of thyrotropin-releasing hormone (TRH, 1-5 microM) on the spontaneous respiratory-related neural activity in a thick brain stem slice preparation from the newborn mouse. The action of TRH on the respiratory output from the slice was investigated by recordings from the XII nerve. Cellular responses to TRH were investigated using whole cell recordings from hypoglossal motoneurons and three types of inspiratory neurons located in the rostral ventrolateral part of the slice. 2. Bath-applied TRH (1 microM) decreased the time between inspiratory discharges recorded on the XII nerve from 12.3 +/- 3.3 s to 4.9 +/- 1.1 s (n = 28; means +/- SD), i.e., caused an approximate threefold increase in the respiratory frequency. The coefficient of variation of the time between the inspiratory discharges decreased by one-half. Thus the respiratory output became more stable in response to TRH. The duration of the inspiratory discharges increased from 474 +/- 108 ms to 679 +/- 114 ms, and the amplitude decreased by 24%. An increase in the interdischarge noise on the XII nerve was recorded in the early phase of the TRH application. 3. Anatomically identified hypoglossal motoneurons (7 cells) responded to bath applied TRH with a depolarization eliciting spikes between the inspiratory potentials. The depolarization was accompanied by an increase in spontaneous excitatory synaptic activity that disappeared late during the TRH application. The duration of the inspiratory potentials was increased, indicating that the hypoglossal motoneurons received a longer duration synaptic input from the respiratory rhythm generator. 4. Type-1 inspiratory neurons showed a prolonged depolarization (3 cells), a transient depolarization (2 cells), or no change in membrane potential (2 cells) during 10 min of continued superfusion with a TRH-containing solution. The duration of the inspiratory potentials was increased during the TRH superfusion. With tetrodoxin (TTX, 1 microM) present in the superfusing solution TRH induced a prolonged depolarization (3 cells) or a transient depolarization (1 cell), demonstrating that type-1 inspiratory neurons are depolarized postsynaptically by TRH. The input resistance was not changed during the depolarizing response to TRH. 5. Type-2 inspiratory neurons showed a transient depolarization (7 cells) in response to bath-applied TRH. The duration of the inspiratory potentials was increased markedly during TRH. The transient depolarization was not the result of a postsynaptic action of TRH, because type-2 neurons (9 cells) showed no depolarization to TRH with TTX present in the superfusing solution. 6. Type-3 inspiratory neurons showed a transient depolarization (4 cells) with a partial recovery of the membrane potential late during the TRH application. The duration of the inspiratory potentials increased markedly during TRH. Four cells showed a transient depolarization with an increase in input resistance during TRH with TTX present in the superfusing solution. Thus type-3 neurons are depolarized postsynaptically by TRH. 7. We conclude that TRH increases the frequency of the respiratory rhythm in newborn mice through an action at the level of the brain stem.(ABSTRACT TRUNCATED AT 250 WORDS) PMID- 8714655 TI - Characteristics of action potentials and their underlying outward currents in rat taste receptor cells. AB - 1. Taste receptor cells produce action potentials as a result of transduction mechanisms that occur when these cells are stimulated with tastants. These action potentials are thought to be key signaling events in relaying information to the central nervous system. We explored the ionic basis of action potentials from dissociated posterior rat taste cells using the patch-clamp recording technique in both voltage-clamp and current-clamp modes. 2. Action potentials were evoked by intracellular injection of depolarizing current pulses from a holding potential of -80 mV. The threshold potential for firing of action potentials was approximately -35 mV; the input resistance of these cells averaged 6.9 G omega. With long depolarizing pulses, two or three action potentials could be elicited with successive attenuation of the spike height. Afterhyperpolarizations were observed often. 3. Both sodium and calcium currents contribute to depolarizing phases of the action potential. Action potentials were blocked completely in the presence of the sodium channel blocker tetrodotoxin. Calcium contributions could be visualized as prolonged calcium plateaus when repolarizing potassium currents were blocked and barium was used as a charge carrier. 4. Outward currents were composed of sustained delayed rectifier current, transient potassium current, and calcium-activated potassium current. Transient and sustained potassium currents activated close to -30 mV and increased monotonically with further depolarization. Up to half the outward current inactivated with decay constants on the order of seconds. Sustained and transient currents displayed steep voltage dependence in conductance and inactivation curves. Half inactivation occurred at 20 +/- 3.1 mV (mean +/- SE) with a decrease of 11.2 +/- 0.5 mV per e-fold. Half maximal conductance occurred at 3.6 +/- 1.8 mV and increased 12.2 +/- 0.6 mV per e-fold. Calcium-activated potassium current was evidenced by application of apamin and the use of calcium-free bathing solution. It was most obvious at more depolarized holding potentials that inactivated much of the transient and sustained outward currents. 5. Potassium currents contribute to both the repolarization and afterhyperpolarization phases of the action potential. These currents were blocked by bath application of tetraethylammonium, which also substantially broadened the action potential. Application of 4-aminopyridine was able to selectively block transient potassium currents without affecting sustained currents. This also broadened the action potential as well as eliminated the afterhyperpolarization. 6. A second type of action potential was observed that differed in duration. These slow action potentials had t1/2 durations of 9.6 ms compared with 1.4 ms for fast action potentials. Input resistances of the two groups were indistinguishable. Approximately one-fourth of the cells eliciting action potentials were of the slow type. 7. Cells eliciting fast action potentials had large outward currents capable of producing a quick repolarization, whereas cells with slow action potentials had small outward currents by comparison. The average values of fast cells were 2,563 pA and 1.4 ms compared with 373 pA and 9.6 ms for slow cells. Current and duration values were related exponentially. No significant difference was noted for inward currents. 8. These results suggest that many taste receptor cells conduct action potentials, which may be classified broadly into two groups on the basis of action potential duration and potassium current magnitude. These groups may be related to cell turnover. The physiological role of action potentials remains to be elucidated but may be important for communication within the taste bud as well as to the afferent nerve. PMID- 8714656 TI - Adaptive control for backward quadrupedal walking V. Mutable activation of bifunctional thigh muscles. AB - 1. In this, the fifth article in a series to assess changes in posture, hindlimb dynamics, and muscle synergies associated with backward (BWD) quadrupedal walking, we compared the recruitment of three biarticular muscles of the cat's anterior thigh (anterior sartorius, SAa; medial sartorius, SAm; rectus femoris, RF) for forward (FWD) and BWD treadmill walking. Electromyography (EMG) records from these muscles, along with those of two muscles (semitendinosus, ST; anterior biceps femoris, ABF) studied previously in this series, were synchronized with kinematic data digitized from high-speed cine film for unperturbed steps and steps in which a stumbling corrective reaction was elicited during swing. 2. During swing, the relative timing of EMG activity for the unifunctional SAm (hip and knee flexor) was similar for unperturbed steps of FWD and BWD walking. The SAm was active before paw lift off and remained active during most of swing (75%) for both forms of walking, but there was a marked decrease in EMG amplitude after paw off during BWD and not FWD swing. In contrast, the relative timing of EMG activity for the SAa and RF, two bifunctional muscles (hip flexors, knee extensors), was different for FWD and BWD swing. During FWD swing, the SAa and the RF (to a lesser extent) were coactive with the SAm; however, during BWD swing, the SAa and RF were active just before paw lift off and then inactive for the rest of swing until just before paw contact (see 3). Thus the swing-phase activity of the SAa and RF was markedly shorter for BWD than FWD swing. 3. Activity in SAa and RF was also different during FWD and BWD stance. The RF was consistently active from mid-to-late stance of FWD walking, and the SAa was also active during this period in some FWD steps. During the stance phase of BWD walking, however, the onset of activity in both muscles consistently shifted to early stance as both muscles became active just before paw contact (the E1 phase). Activity in RF consistently persisted through most of BWD stance. The duration of SAa recruitment during BWD stance was more variable across cats with offsets ranging from mid- to late stance. 4. The activation patterns of the biarticular anterior thigh muscles during stumbling corrective reactions were, in general, similar to their different activations during FWD and BWD swing. The initial response to a mechanical stimulus applied to the dorsum of the paw that obstructed FWD swing was an augmentation of knee flexion and increased activity in ST and SAm. A mechanical stimulus applied to the ventral surface of the paw to obstruct BWD swing resulted in an initial conversion of hip extension to flexion and a slowing of knee flexion. There was a corresponding recruitment of SAa and RF and an enhancement of background activity in SAm. 5. The two forms of walking are differentiated by posture and limb dynamics, yet muscles participating in the basic flexor and extensor synergies are unchanged. Although central pattern generating (CPG) circuits determine the basic timing of these synergies, changes in the duration and waveform of muscle activity may depend on unique interactions among the CPG, supraspinal inputs that set posture and the animal's goal (to walk BWD or FWD) and motion-related feedback from the hindlimb. Output mutability to each muscle may depend on the balance of this tripartite input; muscles with immutable patterns may rely heavily on input from CPG circuits, whereas muscles with mutable patterns may rely more on form-specific proprioceptive and supraspinal inputs. PMID- 8714657 TI - Threshold serotonin concentration required to produce synaptic facilitation differs for depressed and nondepressed synapses in Aplysia sensory neurons. AB - 1. The modulatory actions produced by the neurotransmitter serotonin (5HT) in Aplysia sensory neurons (SNs) can be distinguished on the basis of their concentration requirement for 5HT, their activation and recovery kinetics, and their dependence on the relative contribution of different second messenger pathways. In addition, some of the facilitatory mechanisms activated by 5HT appear to be different depending upon the recent activation history of synaptic transmission from the SNs. In this study, we examined the concentration requirements of 5HT-induced facilitation of depressed and nondepressed synapses. 2. In isolated pleural-pedal ganglia, we produced facilitation of monosynaptic connections between tail SNs and motor neurons (MNs), using different concentrations of 5HT. As a measure of each preparation's greatest sensitivity to 5HT, we first determined the lowest 5HT concentration that produced increased excitability in the SNs ("threshold" 5HT). Then, in one series of experiments, we applied 5HT sequentially to the same synapse, first in the nondepressed and then in the depressed state. In a second series, we applied 5HT simultaneously to two SNs connecting to the same MN; one synapse was depressed, the other nondepressed. 3. In both series of experiments, we found that the 5HT concentration required to produce facilitation of depressed excitatory post-synaptic potentials (EPSPs) was invariably lower than the 5HT concentration that produced facilitation of nondepressed EPSPs. In the first series, 'threshold' 5HT (1.6 microM) was sufficient to facilitate the synapse in the depressed state, but not the nondepressed state. However, the nondepressed synapse could still be facilitated by higher concentrations of 5HT (10 microM). In the second series, increased excitability of SNs, facilitation of depressed synapses, and facilitation of nondepressed synapses were progressively recruited as a function of increasing 5HT concentration (4.1, 6.7, and 10-15 microM, respectively). 4. These data are consistent with previous studies suggesting that different cellular mechanisms contribute to the facilitation of depressed and nondepressed synapses. In addition, our results provide a way to experimentally separate the two processes and to analyze them simultaneously and independently. Taking advantage of this dissociation, in future experiments it may be possible to directly compare the relative contributions of different intracellular mechanisms to synaptic facilitation and to relate them to the degree of recent synaptic activation. PMID- 8714658 TI - Pharmacological and kinetic characterization of two functional classes of serotonergic modulation in Aplysia sensory neurons. AB - 1. Modulation of mechanoafferent sensory neurons (SNs) by the neutrotransmitter serotonin (5HT) plays a significant role in behavioral sensitization of several withdrawal reflexes in Aplysia. The modulatory effects of 5HT on these SNs include increased excitability, increased input resistance, action potential broadening, and increased synaptic transmission. Based on a previously described dissociation of some of these modulatory effects, revealed with the 5HT-receptor antagonist, cyproheptadine, we investigated whether a similar dissociation could be found by systematically varying the concentration of the endogenous agonist, 5HT. 2. We first applied a range of 5HT concentrations to isolated pleural/pedal ganglia (containing tail SNs and tail motor neurons, respectively), and measured the magnitude of 5HT-induced modulation of spike broadening and increased excitability. The resulting dose-response curve showed that both forms of modulation increase monotonically as a function of 5HT concentration, but that excitability has a lower threshold for modulation by 5HT than does spike duration. 3. We further characterized the modulatory effects of 5HT on Aplysia SNs by comparing the time course of onset of modulation by 5HT and the time course of recovery after washout. Independent of 5HT concentration, modulation of excitability increases rapidly in the presence of 5HT and recovers rapidly (< 3 min) after washout. Similarly, input resistance increases and recovers rapidly, mirroring the profile of increased excitability. However, modulation of spike duration exhibits two profiles, dependent on 5HT concentration. Low concentrations of 5HT (0.5 and 1 microM) induce a rapid-onset and transient recovery form of spike broadening, which resembles the kinetics of increased excitability and increased input resistance. Higher concentrations of 5HT (2.5 and 5 microM) induce a more slowly developing and prolonged-recovery form of spike broadening (> 9 min). At these higher concentrations, the recovery profile for prolonged spike broadening is significantly different from those observed for both increased excitability and increased input resistance. 4. We next compared the relationship between spike broadening and short-term synaptic facilitation. We found that significant facilitation of synaptic transmission requires a high 5HT concentration, which is comparable with that required to induce prolonged spike broadening. Similarly, the recovery profiles for spike broadening and synaptic facilitation are strikingly similar, recovering in parallel. 5. Our experiments show that the modulatory effects of 5HT in the tail SNs can be dissociated both by their sensitivity to different concentrations of 5HT and by their kinetics of serotonergic modulation. Based on these results, together with extensive evidence from other laboratories, we propose that the short-term modulatory effects of 5HT fall into two distinct functional classes. The first class, which includes excitability, input resistance, and transient spike broadening, has a low threshold for 5HT modulation and recovers rapidly. The second class, which includes prolonged spike broadening and short-term synaptic facilitation, has a higher threshold for modulation and recovers more slowly. It now will be of interest to determine the functional contribution of each of these classes to different aspects of sensitization. PMID- 8714659 TI - Role of syringeal muscles in gating airflow and sound production in singing brown thrashers. AB - 1. The role of syringeal muscles in song production, particularly in regulating airflow through the syrinx, was studied in singing brown thrashers (Toxostoma rufum). In nine individuals, muscle activity was recorded electromyographically together with bilateral syringeal airflow, subsyringeal air sac pressure, and vocal output. 2. Dorsal muscles, m. syringealis dorsalis (dS) and m. tracheolateral dorsalis (dTB), are consistently activated during ipsilateral closing of the syrinx or increasing syringeal resistance, suggesting that their main role is adduction. This interpretation is supported by the motor patterns accompanying syllables with rapid oscillations in the rate of airflow. Bursts of electrical activity (2-10 ms) in dorsal muscles are precisely synchronized with decreasing airflow. 3. Electrical activity in m. tracheobronchialis ventralis (vTB) and m. tracheolateralis (TL) is associated with active abduction. An important contribution of vTB is to open the syringeal lumen for short inspirations in between syllables. In syllables with oscillatory flow modulations, vTB bursts show variable alignment with the phase of increasing flow. From this and activity during other syllables, it appears that, during phonation, vTB activity fine tunes the syringeal configuration, which is set by action of the dorsal muscles into a partially constricted state. 4. Activity in the ventral portion of TL, an extrinsic muscle, is strikingly similar to that of vTB, an intrinsic muscle, suggesting that the two muscles have a similar functional role. This supports the notion that intrinsic syringeal muscles of songbirds evolved from extrinsic muscles of nonpasserines. 5. M. syringealis ventralis (vS) does not appear to contribute directly to gating of airflow. Its activity is not consistently correlated with active changes in syringeal resistance. 6. Activity in m. sternotrachealis (ST) is most prominent during rapid changes in the rate of airflow or when switching between expiratory and inspiratory flow, suggesting a role in stabilizing the syringeal framework. PMID- 8714660 TI - Low-frequency stimulation at the troughs of theta-oscillation induces long-term depression of previously potentiated CA1 synapses. AB - 1. The induction of long-term weakening of synaptic transmission in rat hippocampal slices was examined in CA1 synapses during cholinergic modulation. 2. Bath application of the cholinergic agonist carbachol (50 microM) activated an oscillation of the local field potential in the theta-frequency range (5-12 Hz), termed theta. It was previously shown that a stimulation train of 40 single shocks (at 0.1 Hz) to the Schaffer collateral-commisural afferents, each synchronized with positive peaks of theta, caused homosynaptic long-term enhancement in CA1. Furthermore, long-term depression (LTD) was sporadically observed when the stimulation train was given at negative troughs of theta. Here we have sought to determine stable conditions for LTD induction during theta. 3. Synaptic weakening was reliably obtained, by giving 40 shocks (at 0.1 Hz) at theta-troughs, only in pathways that had been previously potentiated. This decrement, termed theta-LTD, was synapse specific because it did not occur in an independent pathway not stimulated during theta. The interval between the initial potentiating tetanus and theta-LTD induction could be as long as 90 min. 4. theta LTD could be saturated; after consecutive episodes of theta-LTD induction, no significant further depression was obtained. Moreover, theta-LTD could be reversed by tetanic stimulation. 5. theta-LTD could prevent the induction of LTD by 600-900 pulses at 1 Hz. This suggests that the two protocols may share common mechanisms at the synaptic level. 6. We conclude that single presynaptic spikes that occur at low frequency and are properly timed to the troughs of theta may be a relevant mechanism for decreasing the strength of potentiated synapses. PMID- 8714661 TI - Heterologous modulation of inhibitory synaptic transmission by metabotropic glutamate receptors in cultured hippocampal neurons. AB - 1. Whole cell patch-clamp recordings of monosynaptically connected pairs of hippocampal neurons in very low-density culture were performed to determine the effects of the activation of metabotropic glutamate receptors (mGluRs) on inhibitory terminals. The mGluR agonist (1S, 3R)-1-aminocyclopentane-1,3 dicarboxylic acid [(1S, 3R)-ACPD] and the recently described mGluR antagonist (RS)-alpha-methyl-4-carboxyphenylglycine (MCPG) were used. In addition, the glutamate uptake inhibitors L-trans-pyrrolidine-2,4-dicarboxylate (L-trans-PDC) and D,L-beta-threo-hydroxyaspartate (THA) were used to determine whether endogenous agents (presumably glutamate) could activate mGluRs at inhibitory terminals. Previous reports of the role of mGluRs on inhibitory terminals were performed in slice preparations; our use of patch-clamp recordings from isolated pairs of hippocampal neurons is uniquely useful for the study of inhibitory synaptic transmission in the absence of polysynaptic connectivity. 2. The mGluR agonist (1S, 3R)-ACPD (100 microM) reversibly decreased the amplitude of evoked inhibitory postsynaptic currents (IPSCs) in all pairs tested; this effect was completely blocked by coapplication of the mGluR antagonist MCPG (500 microM) with (1S, 3R)-ACPD. MCPG (500 microM) alone had no effect on IPSC amplitude. These results indicate that all inhibitory neurons in our cultures express functional mGluRs in their terminals. 3. Examination of the frequency and the distribution of amplitudes of miniature IPSCs (mIPSCs) provide indications of changes in the sensitivity of postsynaptic receptors and/or of changes in the process of presynaptic transmitter release. Recordings of miniature currents from hippocampal neurons cultured at very low density makes possible the analysis of mIPSCs that arise from a single input, whereas in high density or slice preparations, spontaneous miniature currents reflect numerous synaptic inputs. No change in the amplitudes or frequency of the mIPSCs were observed upon application of (1S, 3R)-ACPD (100 microM). Thus we conclude that the depression of the evoked IPSC amplitude by (1S, 3R)-ACPD is mediated by a presynaptic mechanism in these isolated pairs of hippocampal neurons. 4. The glutamate uptake inhibitor L-trans-PDC also reduced IPSC amplitude in 8 of 13 pairs. In these eight pairs, an increase in N-methyl-D-aspartate (NMDA) receptor-mediated membrane noise indicated an increase in ambient concentrations of glutamate induced by L-trans-PDC. In the remaining five pairs, membrane noise remained unaffected by L-trans-PDC, and IPSCs were not attenuated. Similar results were observed with the use of the uptake inhibitor THA. The mGluR antagonist MCPG blocked the effects of L-trans-PDC and THA on IPSC amplitude. We propose that inhibition of glutamate uptake mechanisms results in activation of mGluRs on GABAergic terminals via endogenous sources of glutamate and that the uptake inhibitors (L-trans-PDC and THA) do not directly activate the metabotropic receptor. 5. Presynaptic receptors and active modulation of uptake mechanisms are clearly involved in a wide range of physiological and pathological synaptic events. The data presented here suggest that heterosynaptic modulation of inhibitory synaptic transmission by metabotropic glutamate receptors may be important for the maintenance and plasticity of the balances between excitatory and inhibitory synaptic transmission in the CNS. PMID- 8714662 TI - Presynaptic GABAB receptors modulate IPSPs evoked in neurons of deep cerebellar nuclei in vitro. AB - 1. Recording from deep cerebellar nuclei neurons, we investigated the role of presynaptic gamma-aminobutyric acid-B (GABAB) receptors in the modulation of monosynaptic inhibitory postsynaptic potentials (IPSPs) evoked by stimulation of Purkinje cells in rat slice cultures. 2. Bath application of the GABAB receptor agonist, baclofen (10 and 100 microM) induced two effects in cerebellar nuclei neurons: a postsynaptic hyperpolarization of 4.2 +/- 1.7 (SD) mV and a reduction in the amplitude of evoked IPSPs (30 +/- 10%). 3. When the postsynaptic GABAB response was blocked by filling the electrode with cesium methanesulfonate (2 M), or with a solution containing QX 314 (50 mM), bath application of baclofen (10 microM) reversibly depressed the evoked IPSPs by 36.7 +/- 18.7% and 42 +/- 20.3%, respectively. Under these experimental conditions, baclofen (10 microM) also reduced the amplitude of spontaneous IPSPs (10.2 +/- 9.5%) and decreased their frequency by 45.6 +/- 8.8%, suggesting a presynaptic site of action. 4. The presynaptic action of baclofen was not due to activation of receptors on the somata of Purkinje cells: baclofen (100 microM) failed to alter membrane holding current in Purkinje cells, and it had no effect on the rate of spontaneous action potential discharge in Purkinje cells in the presence of ionotropic glutamate receptor antagonists (6-cyano-7-nitroquinoxaline-2,3-dione, 20 microM; D-2-amino 5-phosphonovalerate, 40 microM). 5. IPSPs could be evoked by extracellular stimulation of the Purkinje cell layer or by direct stimulation of the fiber bundle connecting Purkinje cells to deep cerebellar neurons. In both situations, baclofen (10 microM) reduced the amplitude of evoked IPSPs by 32.7 +/- 8.8% and 31.2 +/- 10.2%, respectively. 6. These results demonstrate that GABAB receptors are present on the terminals of Purkinje cells. Their activation causes a decrease in the amplitude of evoked IPSPs recorded in deep cerebellar nuclei and also reduces the frequency of spontaneous inhibitory events. PMID- 8714663 TI - Contribution of GABA- and glycine-mediated inhibition to the monaural temporal response properties of neurons in the inferior colliculus. AB - 1. To determine the contribution of inhibition to the generation of the temporal response patterns of neurons in the inferior colliculus (IC), the effects of iontophoretically applied gamma-aminobutyric acid (GABA), glycine, and the GABAA and glycine receptor antagonists, bicuculline and strychnine were studied on 121 neurons in the IC of urethan-anesthetised guinea pig. 2. The neurons temporal discharge patterns were classified into six categories on the basis of their peristimulus time histograms (PSTHs). 1) Onset units fired at the stimulus onset and could be divided into two subtypes: narrow (1-2 spikes only) or broad (response lasting up to approximately 30 ms). 2) Pauser units had a precisely timed onset peak separated from a lower level of sustained activity by either a marked reduction or complete cessation of firing. 3) Chopper units had three or more clearly defined peaks near stimulus onset or evidence of regularly spaced peaks over the duration of the stimulus. 4) Onset-chopper units had three clearly defined peaks at onset but no sustained firing. 5) On-sustained units had a clearly defined single onset peak followed by a lower level of sustained activity. 6) Sustained units fired throughout the stimulus, but lacked an onset peak. 3. Iontophoretic application of GABA and glycine produced a dose-dependent reduction in firing rate in 76% (42/55) and 79% (11/14) of units, respectively. Application of bicuculline or strychnine increased the discharge rate in 91% (64/70) and 94% (16/17) of neurons, respectively. 4. The effects of bicuculline and strychnine on PSTH class were studied in detail on 70 neurons. Changes in discharge rate were accompanied by changes in PSTH in 49% (34/70) of neurons tested with bicuculline and 41% (7/17) tested with strychnine. Pauser units were the most affected with 69% changing their PSTH class, but some units in all PSTH classes, except the chopper group, exhibited changes in PSTH pattern after application of bicuculline. The majority of units (approximately 50%) that changed PSTH pattern in the presence of bicuculline became chopper units. Units of all PSTH classes could become choppers, but the proportion of units showing this change was dependent on the unit's control response pattern. All seven units that changed PSTH class with strychnine also became choppers. Changes in PSTH, including the appearance of a chopper pattern, did not depend on either a unit's control discharge rate or the magnitude of the change in discharge rate induced by the antagonists. 5. Bicuculline and strychnine had no significant effect on latency for units in the chopper, onset-chopper, onset, pauser, and on-sustained groups. A few sustained and unclassified units that had long predrug latencies did show marked reductions in latency when tested with bicuculline. 6. The majority of units did not fire spontaneously, and neither bicuculline or strychnine produced a significant increase in spontaneous rate. 7. In many units, the changes in firing rate did not occur equally over the duration of the response. Firing rates at the onset and in the last quarter of the sustained response were compared. Three effects of bicuculline and strychnine were observed. For 80% of units the largest change in firing rate occurred in the sustained response, while in 14% of units the change was greatest at onset. PMID- 8714664 TI - Importance of NMDA receptors for multimodal integration in the deep layers of the cat superior colliculus. AB - 1. Many sensory events contain multimodal information, yet most sensory nuclei are devoted to the analysis of single-modality information. In the deep superior colliculus (DSC), visual, auditory, and somatosensory information converges on individual multimodal neurons. The responses of multimodal neurons are determined by the temporal and spatial correspondence properties of the converging inputs such that stimuli arising from the same event elicit a facilitated multimodal response. 2. N-methyl-D-aspartate (NMDA) receptors may underlie the detection of spatial and temporal coincidence and could be involved in the generation of multimodal facilitatory responses because of the nonlinear properties of NMDA receptor-mediated events. To assess the role of NMDA receptors in multimodal integration, we made extracellular recordings from single multisensory neurons in the DSC of the cat. 3. The responses to visual, auditory, and somatosensory stimuli alone and to multimodal combinations of stimuli were challenged with iontophoretically applied D-2-amino-5-phosphonovalerate (AP5), an NMDA receptor antagonist. All responses to visual stimuli presented alone (n = 9) were greatly reduced. Somatosensory responses (n = 25) were usually decreased. In contrast, the responses to auditory stimulation were decreased (n = 9), unaffected (n = 3), or enhanced (n = 5). 4. Responses to multimodal stimulus presentations were consistently reduced during iontophoretic application of AP5, irrespective of the modalities that made up the stimulus. The reductions of multimodal responses were significantly greater than the sum of the reductions of responses to single modality stimuli. 5. The data suggest that for unimodal stimuli, the importance of NMDA receptors in synaptic transmission of sensory responses in DSC may be dependent on the stimulus modality. Furthermore, NMDA receptors are of major importance in the integration of input from different modalities for the generation of multimodal responses. PMID- 8714665 TI - Electrical properties of neocortical neurons in slices from children with intractable epilepsy. AB - 1. The intrinsic electrical properties of human neocortical neurons were studied with current-clamp and single-electrode voltage-clamp techniques in slices obtained from children, aged 3 mo to 15 yr, undergoing surgical treatment of intractable epilepsy. Neocortical samples were classified as most or least abnormal based on clinical data. Recorded neurons were labeled with biocytin for correlation of electrical properties with morphological characteristics and laminar position. All recorded neurons were divided into three cell types--fast spiking, low-threshold spiking (LTS) and non-LTS cells--on the basis of their electrical characteristics. 2. Fast-spiking cells generated brief, rapidly repolarizing action potentials. Most of these cells showed only weak spike frequency adaptation. Fast-spiking cells labeled with biocytin were aspiny or sparsely spiny nonpyramidal neurons located in cortical layers 2-4. 3. LTS cells generated Ca(2+)-dependent low-threshold potentials and were the most numerous of the three cell types. Their Na(+)-dependent action potentials were broader than those of fast-spiking cells and showed marked spike-frequency adaptation. The size of low-threshold Ca2+ potentials and currents varied across cells, but they never supported more than two or, occasionally, three fast action potentials. LTS cells were pyramidal neurons located throughout cortical layers 2-6. Unlike the bursting neocortical cells described in lower mammals, LTS neurons in neocortex from children failed to generate bursts of inactivating Na+ action potentials. 4. Non-LTS cells also had relatively broad Na(+)-dependent action potentials and showed spike-frequency adaptation, but they did not generate detectable low threshold potentials or currents. Non-LTS cells were also pyramidal neurons located throughout layers 2-6. 5. The electrical properties of cells from different age groups (< or = 1, 2-8, and 9-15 yr) and from most-abnormal and least-abnormal tissue samples were compared. A statistically significant trend toward a lower input resistance, a faster membrane time constant, and a decreased spike duration was observed with increasing age. There were no significant differences between the electrical properties of cells from the most-abnormal tissue and cells from the least-abnormal tissue. 6. These data indicate that the intrinsic electrical properties of neocortical neurons from children vary according to cell morphology and change with increasing age, as has been observed in rodent and feline neocortical neurons. No obvious evidence of epileptogenicity was detected in the intrinsic electrical properties of any of the neurons studied. PMID- 8714666 TI - Neural mechanisms of blood flow regulation during synaptic activity in cerebellar cortex. AB - 1. We investigated the neural mechanisms of the increases in blood flow produced by synaptic activity using the parallel fiber (PF) system of the cerebellum as a model. The midline cerebellum was exposed in anesthetized rats and the PFs were stimulated with tungsten microelectrodes. Cerebellar blood flow (BFcrb) was recorded using a laser-Doppler probe, whereas field potentials were recorded using glass micropipettes. PF stimulation produced increases in BFcrb that were related to the frequency and intensity of stimulation (+60 +/- 9%, mean +/- SE, at 100 microA and 30 Hz; n = 6). The greatest increases were confined to a band stretching along the major axis of the stimulated folium and corresponding to the beam of activated PFs. The increase in evoked by PF stimulation was associated with a corresponding increase in glucose utilization, assessed by the 2 deoxyglucose method. The increases in BFcrb and the field potentials evoked by PF stimulation were abolished by tetrodotoxin (1 microM; n = 6). Ringer solution containing 12 mM Mg2+ and 0 mM Ca2+ blocked synaptic activity in the PFs and abolished the increases in flow (P > 0.05 from baseline; n = 5). The broad spectrum glutamate receptor antagonist kynurenate (5 mM) prevented depolarization of Purkinje cells and interneurons and abolished the increase in BFcrb evoked by PF stimulation (P > 0.05; n = 6). Treatment with tetrodotoxin, Mg2+, or kynurenate did not affect the increase in BFcrb elicited by systemic hypercapnia or by topical application of the nitric oxide donor 3-morpholino sydnonimine (P > 0.05 from Ringer solution). We conclude that the increases in flow produced by synaptic activity are linked to glutamate-induced depolarization of Purkinje cells and interneurons. These findings provide evidence that activation of glutamate receptors participates in the mechanisms of functional hyperemia, and they support the validity of the PF system as a model for study of the relationship between synaptic activity and blood flow in the CNS. PMID- 8714667 TI - Two types of network oscillations in neocortex mediated by distinct glutamate receptor subtypes and neuronal populations. AB - 1. Two distinct forms of spontaneous synchronous oscillations were investigated with field potential recordings in slices of rat somatosensory cortex in vitro. 2. The first type of synchronous oscillation was activated by low extracellular [Mg2+] and had dominant frequencies of 8-12 Hz. It was abolished reversibly by the N-methyl-D-aspartate (NMDA) receptor antagonists D-2-amino-5-phosphonovaleric acid and was relatively unaffected by the non-NMDA receptor antagonist 6,7 dinitroquinoxaline-2,3-dione (DNQX). The duration of oscillatory events was increased by blocking gamma-aminobuturic acid-A receptors with bicuculline or by activating metabotropic glutamate receptors with trans-1-aminocyclopentane-1,3 dicarboxylic acid. 3. A second form of synchronous oscillation was activated by acute application of kainic acid (10 microM), had dominant frequencies of 1-5 Hz, and was abolished reversibly by DNQX. Low concentrations of domoic acid mimicked the effects of kainate, but alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid or quisqualic acid did not, suggesting a role for the GluR5-7 and KA1-2 glutamate receptor subunits. 4. Surgical isolation of cortical layers showed that spontaneous NMDA receptor-dependent oscillations originated within layer 5 exclusively, but kainate receptor-dependent oscillations were uniquely generated by neurons in layers 2/3. 5. Our results suggest that neocortical neurons in layers 2/3 and layer 5 can independently generate two distinct forms of rhythmic population activity, each dependent upon activation of a different subtype of glutamate receptor. PMID- 8714668 TI - Bistability and its regulation by serotonin in the endogenously bursting neuron R15 in Aplysia. AB - 1. Previous computational studies of models of neuron R15 in Aplysia have indicated that several distinct modes of electrical activity may coexist at a given set of parameters, that this multistability can be modulated by transmitters such as serotonin (5-HT) and that brief perturbations of the membrane potential can induce persistent changes in the mode of electrical activity. To test these predictions, the responses of R15 neurons to injections of brief (1.5 s) current pulses were recorded intracellularly in the absence and presence of 5-HT. 2. In the absence of 5-HT, brief perturbations induced abrupt transitions in the electrical activity from bursting to beating. Such transitions were observed in approximately 20% of the cases. The duration of beating activity varied from several seconds to tens of minutes. In the presence of low concentrations (1 microM) of 5-HT, both the probability of mode transitions and the duration of induced beating activity increased significantly. 3. These results indicate that at least two stable modes of electrical activity can coexist in R15 neurons and that this bistability can be regulated by 5-HT. In general, these conclusions agree with the results from analyses of mathematical models of R15. Although the function of these dynamic properties in R15 is speculative, our results, interpreted on the background of the model, support the notion that nonlinear dynamical properties of individual neurons can endow them with richer forms of information processing than has generally been appreciated. PMID- 8714670 TI - Alterations in glomerular permeability in streptozotocin-induced diabetic rats. AB - The alteration in glomerular basement membrane permeability associated with microangiopathy in streptozotocin-induced diabetic rats was studied by determining the movement across the glomerular basement membrane of anionic ferritin probes injected into rats at different points in the development of the disease. Visualization of the concentration gradient of anionic ferritin and changes in ultrastructure was accomplished by electron microscopic examination of renal tissue prepared from both control and diabetic rats. In all control rats, the anionic ferritin did not leave the glomerular capillary lumen, nor were there any changes in the normal morphology of the glomerular capillary wall. In the diabetic animals, the concentration of anionic ferritin shifted from the capillary lumen in the abluminal direction. Distinct morphologic changes, such as widening of endothelial intercellular junctions, focal detachment of podocyte foot processes, and extensive thickening of the glomerular basement membrane, were noted in the diabetic rat, and these changes appear to correlate with the observed increase in permselectivity of anionic ferritin across the glomerular capillary wall. PMID- 8714669 TI - Cellular short-term memory from a slow potassium conductance. AB - 1. We use the dynamic clamp to add the slowly inactivating and slowly recovering K+ conductance Kv1.3 to cultured stomatogastric ganglion neurons. 2. Introduction of Kv1.3 produced long delays to firing during depolarization. Additionally, the slow recovery from inactivation produced an increase in neuronal excitability after a depolarizing input that outlasted the input by many seconds. Finally, when introduced into bursting neurons, Kv1.3 produced a long-lasting depolarization-induced switch between tonic and burst firing. 3. These data demonstrate that the slow kinetics of a K+ conductance can produce a form of cellular short-term memory that is independent of any changes in synaptic efficacy. PMID- 8714671 TI - Tailor's bunion. Is fixation necessary? AB - Tailor's bunion or bunionette are terms that describe a pathologic enlargement occurring laterally on the fifth metatarsophalangeal joint. Regardless of the etiology that precipitates the deformity, the resulting abnormal protrusion of soft tissue or bone can result in pain for the patient. Symptoms can range from mild discomfort to severe, debilitating pain. The patient may present with pain dorsolaterally, laterally, or plantarly. The symptoms are mechanically induced, and are often associated with hyperkeratotic lesions and adventitious bursae. Patients complain most often that they cannot find comfortable shoes. The authors compare the effectiveness of fixated versus nonfixated distal osteotomies of the fifth metatarsal for the correction of tailor's bunion. This study shows that fixation can help control postoperative dorsal displacement of the fifth metatarsal capital fragment (p < 0.0001) and produce less shortening of the metatarsal resulting in fewer complications. PMID- 8714672 TI - Integer-driven relational database for clinical outcomes research. AB - A working database program specifically designed to organize and analyze data for the purpose of generating clinical outcomes reports is presented. Following the author's hypothesis, the database accepts only integer input but can generate reports in textual format. The pilot model database presented here uses only five clinic data items, three outcomes items, and can generate 90 different outcomes reports in the form of charts, graphs, or grids. The program consists of a graphical user interface front end that drives a relational database. PMID- 8714674 TI - Foot pain and disability in older persons. Pilot study in assessment and education. PMID- 8714673 TI - Tarsal tunnel syndrome. A retrospective study. AB - Many etiologies of tarsal tunnel syndrome exist, but it is difficult to determine the specific etiology for a patient. Surgical success rates vary in the literature from 44% to 100%. This retrospective study reviewed 40 patients with 45 data sets. Thirty-five patients (87.5%) in the study were female; 35 patients (87.5%) were caucasian. Thirty-two feet (71.1%) showed improvement following the tarsal tunnel surgery. The Tinel's sign parameter improvement was statistically significant. The differences in the subjective parameters of burning, radiating, and sharp sensations were statistically significant, while the differences in pins and needle sensation were nearly significant. The Takakura index and its constituent parameters were all statistically significant postoperatively with the exception of muscle atrophy, which did not have sufficient data for testing. PMID- 8714675 TI - Tumoral calcinosis of the plantar forefoot: a case study. PMID- 8714676 TI - Neurofilament phosphorylation. AB - Neurofilament proteins (NFPs) are highly phosphorylated molecules in the axonal compartment of the adult nervous system. The phosphorylation of NFP is considered an important determinant of filament caliber, plasticity, and stability. This process reflects the function of NFs during the lifetime of a neuron from differentiation in the embryo through long-term activity in the adult until aging and environmental insult leads to pathology and ultimately death. NF function is modulated by phosphorylation-dephosphorylation in each of these diverse neuronal states. In this review, we have summarized some of these properties of NFP in adult nervous tissue, mostly from work in our own laboratory. Identification of sites phosphorylated in vivo in high molecular weight NFP (NF-H) and properties of NF-associated and neural-specific kinases phosphorylating specific sites in NFP are described. A model to explain the role of NF phosphorylation in determining filament caliber, plasticity, and stability is proposed. PMID- 8714677 TI - A role for neurofilaments in the pathogenesis of amyotrophic lateral sclerosis. AB - Amyotrophic lateral sclerosis (ALS) is a late-onset degenerative disease of motor neurons, characterized by abnormal accumulation of neurofilaments (NFs) in perikarya and proximal axons. Two lines of evidence suggest that neurofilament accumulation can play a crucial role in ALS pathogenesis. First, transgenic mouse models overexpressing NF proteins were found to develop motor neuron degeneration and, second, variant alleles of the NF heavy-subunit (NF-H) gene have been found in some human ALS patients. Our axonal transport studies with transgenic mice overexpressing the human NF-H gene, a model of ALS, revealed defects of intracellular transport not only for neurofilament proteins but also for other cytoskeletal proteins and organelles such as mitochondria. Therefore, we propose that neurofilament accumulation in mice causes neurodegeneration by disrupting axonal transport, a mechanism that may account for the pathogenesis of ALS. PMID- 8714678 TI - Changes in retinal ganglion cell axons after optic nerve crush: neurofilament expression is not the sole determinant of calibre. AB - After injury in the central nervous system of adult mammals, many of the axons that remain attached to their intact cell bodies degenerate and decrease in calibre. To understand this process better, we have investigated the relationship between axonal loss, cell loss, and the time course of changes in axonal calibre. Optic nerves (ONs) were crushed and the numbers and sizes of axons remaining close to the cell bodies (2 mm from the eye) and near the site of the lesion (6 mm from the eye) were determined for nerves examined between 1 week and 3 months after injury. Comparison with the retinal ganglion cell (RGC) counts from the same animals revealed that axonal loss was concomitant with cell body loss for at least the first 2 weeks after injury. However, there was no significant change in the calibre of the surviving neurons until 1 month after injury. Thereafter, the axonal calibre was decreased equally along the ON. No progressive somatofugal atrophy was observed. These decreases in axonal calibre occur much later than the immediate drop in neurofilament (NF) expression that also follows injury. The late effect of injury on axonal calibre suggests that NF expression is not the sole determinant of axon size of the RGC fibers in the ON. Other factors are likely additional contributing factors, such as the decreased rate of axonal transport that would help maintain the axonal neurofilament content. PMID- 8714679 TI - Dystonin transcripts are altered and their levels are reduced in the mouse neurological mutant dt24J. AB - Dystonia musculorum is a hereditary mouse neurodegenerative disorder that primarily affects the sensory arm of the nervous system. We have recently cloned and identified a candidate gene for this disorder and designated it dystonin. The sequence of dystonin predicts a rod-shaped cytoskeletal-associated protein with an actin-binding domain at the N-terminal end and a hemidesmosomal protein sequence (bpag1) at the C-terminal end. Here we show that abnormal dystonin transcripts are present in neural tissues of a spontaneous dystonia musculorum mutant, dt24J. We further show that dystonin transcript levels are reduced 2- to 3-fold in dt24J mice. PMID- 8714680 TI - Overexpression of human keratin 16 produces a distinct skin phenotype in transgenic mouse skin. AB - Human cytokeratin 16 (K16; 48 kDa) is constitutively expressed in postmitotic keratinocytes in a variety of stratified epithelial tissues, but it is best known for the marked enhancement of its expression in stratified squamous epithelia showing hyperproliferation or abnormal differentiation. Of particular interest to us, K16 is strongly induced at the wound edge after injury to the epidermis, and its accumulation correlates spatially and temporally with the onset of reepithelialization. To examine the properties of K16 in its natural cellular context, we introduced a wild-type human K16 gene into the germ line of transgenic mice. Several transgenic lines were established and characterized. Under most conditions, the human K16 transgene is regulated tissue specifically in the skin of transgenic mice. Animals that feature low levels of transgene expression are indistinguishable from controls during the first 6-8 months of life. In contrast, transgenic animals expressing the transgene at higher levels develop skin lesions at 1 week after birth, coinciding with the emergence of fur. At a cellular level, alterations begin with the reorganization of keratin filaments and are first seen at the level of the hair follicle outer root sheath (ORS), where K16 expression is known to occur constitutively. The lesions then progressively spread to involve the proximal epidermis, with which the ORS is contiguous. Elevated transgene expression is associated with a marked thickening of these two epithelia, along with altered keratinocyte cytoarchitecture and aberrant keratinization but no keratinocyte lysis. The implications of this phenotype for epithelial differentiation, human genodermatoses, and wound healing in skin are discussed. PMID- 8714681 TI - Cytokeratin expression, fibrillar organization, and subtle function in liver cells. AB - Cytokeratins (CKs) constitute a diverse group of intermediate filament (IF) proteins, expressed as pairs in keratinized and nonkeratinizing epithelial cells. Much is known now about the expression, assembly, and function of CKs in keratinized epithelial cells, the main features being the tight coupling between CK pair switch and cell terminal differentiation (protection barrier) and the vital role of CK IFs in cell mechanical integrity. However, the picture about nonkeratinizing epithelia, like the hepatic tissue, remains quite unclear. The liver forms a multicellular system, where parenchymal cells (i.e., hepatocytes) exert diverse metabolic function(s) and nonparenchymal epithelial cells (e.g., biliary epithelial cells) usually serve structural (or accessory) purposes. In terms of differential CK gene expression, the data accumulated so far demonstrated that parenchymal cells can contain as few as one single CK pair, whereas nonparenchymal cells contain more than two CKs, one of them being a representative of those found in epidermis. Moreover, the distribution of the CK IF networks present in the different cell types varies a lot and can often be linked to the cell specialization. However, the function(s) played by these IF proteins in this multicellular tissue remains a major issue. The use of new experimental approaches, largely based on gene transfer technology, indicates that it is quite subtle. PMID- 8714682 TI - Alterations of intermediate filaments in various histopathological conditions. AB - Intermediate filament proteins belong to a multigene family and constitute an important cytoskeletal component of most vertebrate cells. Their pattern of expression is tissue specific and is highly controlled during embryonic development. Numerous pathologies are known to be associated with modifications of intermediate filament organisation, although their precise role has not yet been elucidated. The present review focuses on the most recent data concerning the possible causes of intermediate filaments disorganization in specific pathologic conditions affecting the epidermis, the liver, and the nervous system. We discuss the formation of abnormal intermediate filament networks that arise as a consequence of mutations that directly affect intermediate filament structure or are induced by multifactorial causes such as modifications of post translational processes and changes in the levels of expression. PMID- 8714683 TI - Localization of Merkel cells at hairless and hairy human skin sites using keratin 18. AB - Merkel cells are neurosecretory cells of the skin with epithelial features such as desmosomes and expression of keratins 8, 18, 19, and 20. Merkel cells are scarcely distributed in adult human skin. Although they are present in hair follicles, their density is higher at hairless anatomic sites such as palms and soles. These cells are often innervated by sensory nerve fibers and are thought to be specialized mechanosensory skin receptor cells. However, their precise origin and function are not clearly established. The aim of this study was to localize Merkel cells in human hairless and hairy skin by immunohistochemistry with antibodies Ks18.174 and Ks19.1 directed against keratins 18 and 19, respectively. In glabrous skin of palm and sole, Merkel cells have been localized at the bottom of the rete ridges, in the epidermal basal layer. To study Merkel cell distribution at hairy anatomic sites, we have chosen breast skin, a tissue containing small hair follicles typical of those covering most of the body's surface. Merkel cells were present in the interfollicular epidermis. In hair follicles, they have been identified in the isthmus region. PMID- 8714684 TI - Biliary secretion and actin-cytokeratin filament distribution in rat hepatocytes during phalloidin-induced cholestasis. AB - The relationship between bile secretion (bile flow, bile acids, phospholipids, and cholesterol) and distribution of actin microfilaments (MFs) and cytokeratin (CK) intermediate filaments (IFs) was examined in hepatocytes of rats injected with a single low dose of phalloidin. This treatment induced a transient cholestasis characterized by a rapid development period (0-90 min postinjection) and a slow recovery period (24 h and 5 days postinjection). No significant changes were observed in bile acid secretion during the 5-day period. The phospholipid output dropped to less than 25% at 90 min and was back to the normal value at 24 h postinjection. In a parallel way, the cholesterol secretion dropped to 30% but came back to only 60% of the control level. Nile Red staining demonstrated a concomitant accumulation of lipids both in the cytoplasm and at the surface membrane. Immunostaining of the actin MFs and CK IFs showed that, in contrast with controls where both cytoskeletal networks were preferentially and uniformly localized at the surface membrane (i.e., sinusoidal, basolateral, and canalicular regions), the toxin treatment led to a major targeting of actin to the pericanalicular region at 24 h and a massive accumulation of well-preserved CK IFs in the cytoplasm at 5 days. Interestingly, this accumulation of CK IFs was not linked to any significant variations in CK isoforms. Together, these data indicate that a selective binding of the toxin to sinusoidal membrane actin at the time of injection triggers a sequence of events that culminate in delayed accumulation of actin MFs at the canalicular pole and of CK IFs in the cytoplasm. Moreover, the reversible perturbation of the bile secretory activity implies a functional adaptation of the hepatocytes that parallels the phalloidin-induced reorganization of both cytoskeleton networks. PMID- 8714685 TI - Purification of developmentally regulated avian 400-kDa intermediate filament associated protein. Molecular interactions with intermediate filament proteins and other cytoskeleton components. AB - IFAPa-400, a 400-kDa developmentally regulated protein thought to be associated with intermediate filaments, has been purified from chick embryo hearts to investigate its interaction with vimentin and other IF proteins and to identify other cellular components to which this cytoskeletal protein associates. Previous studies suggested that this protein was associated with the vimentin-containing intermediate filament lattice of myoblasts and neuroblasts before their terminal differentiation, providing these cells with a particular intermediate filament cytoskeleton that could satisfy specific mechanical requirements during their intense morphogenetic activities. Although IFAPa-400 partially reassociated with vimentin and desmin in disassembly-reassembly experiments using crude IF preparations from chick embryo hearts, in vitro recombination of purified IFAPa 400 with vimentin and desmin failed to demonstrate any direct association. When purified IFAPa-400 was used as a probe in blot overlay assays, however, specific binding to vimentin and desmin was observed, providing the first evidence of a physical association between IFAPa-400 and intermediate filament proteins. The blot overlay experiments also demonstrated that IFAPa-400 binds to two unidentified polypeptides of 19 and 32 kDa. These results are thus consistent with the hypothesis that a structural lattice requiring a vimentin-IFAPa-400 combination constitutes the intermediate filament system of myogenic and neurogenic cells. PMID- 8714686 TI - Tubulin expression and axonal transport in injured and regenerating neurons in the adult mammalian central nervous system. AB - Microtubules are essential components of the cytoskeleton required for axonal growth. To investigate how changes in tubulin transport and expression may affect axon regeneration, injury in the adult mammalian central nervous system was studied. Axotomized retinal ganglion cells (RGCs) that do not regenerate were compared with RGCs that regenerate their axons when the optic nerve is replaced with a peripheral nerve graft. When RGC axons regenerated through peripheral nerve grafts, the rate of slow transport increased but decreased when no regrowth occurred. To investigate the molecular mechanisms that mediate these responses, alterations in tubulin mRNA levels after injury were examined. Total tubulin mRNA levels fell after injury in the optic nerve but increased in those RGCs that regenerated their axons into a peripheral nerve graft. Further, the expression of four separate beta-tubulin isotypes in injured rat RGCs was characterized. mRNA levels for all four isotypes decreased in RGCs after injury in the optic nerve. How the autoregulation of tubulin expression may contribute to the changes in beta-tubulin isotype expression after injury is discussed. PMID- 8714687 TI - Mechanics of motility: distinct dynein binding domains on alpha- and beta tubulin. AB - Microtubules (MTs) interact with force-generating proteins to generate a variety of intracellular movements, including intracellular particle transport, ciliary flagellar beating, and chromosome-spindle movements during mitosis-meiosis. Relatively little is known about the mechanics of these motor-MT interactions, in part because the motor binding domains of the MT and the corresponding MT binding domains of the motor have not been well characterized. Using a flagellar motility assay, we report that the MT subunits, alpha- and beta-tubulin, each contain a dynein binding domain located near the C-termini of their respective tubulin subunits. Blocking either alpha- or beta-tubulin binding domains of dynein attenuates motility in demembranated sea urchin sperm up to 50%. Interestingly, blocking both alpha- and beta-tubulin binding domains on dynein produces much greater decreases in motility. These data suggest that flagellar dynein binds to both subunits of the MT polymer, alpha- and beta-tublin. In addition, the two subunits appear to contribute equivalent, but functionally separate, roles to flagellar motility. PMID- 8714688 TI - Production and utilization of detyrosinated tubulin in developing Artemia larvae: evidence for a tubulin-reactive carboxypeptidase. AB - The reversible, enzymatically driven removal and readdition of its carboxy terminal tyrosine are major posttranslational modifications of alpha-tubulin. To study these processes isoform-specific antibodies were produced and subsequently used to characterize tyrosinated and detyrosinated tubulin in the brine shrimp, Artemia. Tyrosinated tubulin existed in relatively constant amounts on western blots of cell-free protein extracts from Artemia at all developmental stages examined, whereas detyrosinated tubulin was present after 20-24 h of postgastrula growth. In agreement with the blots, the detyrosinated isoform was observed in immunofluorescently stained larvae after 24 h of incubation, appearing first in structures of a transient nature, namely spindles and midbodies. The elongated muscle cells encircling the gut and the epithelium bordering the gut lumen were stained extensively with antibody to detyrosinated tubulin. Detyrosination was accompanied by the appearance of a tubulin-reactive carboxypeptidase, which used both nonpolymerized and polymerized tubulin as substrate. The enzyme bound to microtubules very poorly, if at all, under conditions used in this work. Several inhibitors of carboxypeptidase A had no effect on the carboxypeptidase from Artemia and revealed similarities between this enzyme and others thought to be tubulin specific. The use of inhibitors also indicated that the carboxypeptidase from Artemia recognized aspects of tubulin structure in addition to the carboxy terminal tyrosine. Our results support the idea that detyrosinated tubulin appears in microtubules of varying stability, and they demonstrate that Artemia possess a carboxypeptidase with the potential to detyrosinate tubulin during growth of larvae. PMID- 8714689 TI - Effects of taxol on the polymerization and posttranslational modification of class III beta-tubulin in P19 embryonal carcinoma cells. AB - Undifferentiated P19 embryonal carcinoma cells and P19 cells induced to differentiate along a neuronal pathway by 10(-6) M retinoic acid were treated with taxol to examine the effects of this microtubule-stabilizing drug on the subcellular sorting of class III beta-tubulin and on neurite outgrowth. P19 cells were grown on cover slips and then treated with taxol at concentrations of 10(-6) to 10(-9) M for 24 h. The microtubule cytoskeleton was examined after double immunofluorescence labelling with a monoclonal antibody to alpha-tubulin (YOL 1/34) and a monoclonal neuron-specific class III beta-tubulin antibody (TuJ1). Treatment of undifferentiated P19 cells with concentrations of taxol greater than 4 x 10(-8) M caused microtubule bundling and multiple aster formation and promoted polymerization of the low levels of class III beta-tubulin found in these cells. In neurons, at 2 x 10(-8) M taxol, bundling of microtubules at the base of the neurite was apparent. At taxol concentrations greater than 1 x 10(-7) M, enhanced assembly of class III beta-tubulin was apparent, although long neurites were not observed. Using isoelectric focusing followed by western blotting, we detected an additional isoform of class III beta-tubulin after treatment with 10(-6) M taxol. The results indicate taxol treatment alters the normal subcellular sorting of tubulin isotypes, promotes the polymerization and posttranslational modification of class III beta-tubulin, and interferes with neurite outgrowth. PMID- 8714690 TI - Accumulation of microtubule-associated protein 1A (MAP1A) in differentiating P19 embryonal carcinoma cells. AB - We have examined the accumulation of MAP1A in retinoic acid induced P19 embryonal carcinoma (EC) neurons. By immunofluorescent confocal microscopy, MAP1A was detected in the mitotic spindle of undifferentiated cells but was not evident in association with the interphase microtubules in most cells. By day 4 of differentiation, when neurite outgrowth was underway, MAP1A was co-localize with microtubules in all neurites but was absent from growth cones. By day 8, substantial neurite outgrowth had occurred and MAP1A was seen in all processes. At day 12, no further neurite outgrowth was evident and existing neurites were organized into fascicles. Western blotting and ELISA showed that MAP1A protein levels increased during differentiation. Peak accumulation occurred no later than day 8, coinciding with the period of neurite outgrowth, and then decreased after day 8. The results suggest that in differentiating P19 EC cells MAP1A modulates microtubule dynamics during neurite outgrowth. PMID- 8714691 TI - Modulation of actin dynamics during stress and physiological stimulation by a signaling pathway involving p38 MAP kinase and heat-shock protein 27. AB - HSP27, like other proteins of the heat-shock protein family, accumulates to high levels after exposure of cells to a short period of hyperthermia and contributes to the development of a transient state of thermoresistance. In vitro, HSP27 behaves as an actin cap-binding protein and can inhibit actin polymerization. In vivo, the protective function of HSP27 is exerted mainly at the level of the microfilaments and appears as an extension of a normal function of the protein. This function is regulated by phosphorylation in a mitogen- and stress-sensitive signaling pathway involving the newly characterized p38 MAP kinase. The phosphorylation-modulated function of HSP27 can contribute to agonist-induced reorganization of the actin cytoskeleton and, in the case of stress activation, provides an actin-based adaptive response of cells to the new environmental conditions. PMID- 8714692 TI - Microfilament organization and wound repair in retinal pigment epithelium. AB - Several systems of microfilaments (MF) associated with adherens-type junctions between adjacent retinal pigment epithelial (RPE) cells and between these cells and the substratum play an important role in maintaining the integrity and organization of the RPE. They include prominent, contractile circumferential MF bundles that are associated with the zonula adherens (ZA) junctions. In chick RPE, these junctions are assembled from smaller subunits thus giving greater structural flexibility to the junctional region. Because the separation of the junctions requires trypsin and low calcium, both calcium-dependent and independent mechanisms are involved in keeping adjacent RPE cells attached to one another. Another system of MF bundles that crosses the cell at the level of ZA junctions can be induced to form by stretching the epithelium. The MF bundles forming this system are oriented in the direction in which the RPE is stretched, thereby preventing the overextension of the cell in any one direction. The system may be useful as an indicator of the direction in which tension is experienced by RPE during development of the eye, in animal models of disease and during repair of experimentally induced wounds. Numerous single-cell wounds resulting from death of RPE cells by apoptosis at various stages of repair are normally present in developing chick and adult mammalian RPE. These wounds are repaired by the spreading of adjacent RPE cells and by the contraction of MF bundles oriented parallel to the wound edge, which develop during this time. As a result of the spreading in the absence of cell proliferation, the RPE cells increase in diameter with age. Experimentally induced wounds made by removing 5-10 RPE cells are repaired by a similar mechanism within 24 h. In repair of larger wounds, over 125 microns in width, the MF bundles oriented parallel to the wound edge characteristic of spreading cells are later replaced by stress fibers (SFs) that run perpendicularly to the wound edge and interact with the substratum at focal contacts (FCs) as RPE cells start to migrate. Cell proliferation is induced in cells along the wound edge only when the wounds are wide enough to require cell migration. In the presence of antibodies to beta-1-integrins, a component of FCs, cell spreading is not prevented but both cell migration and cell proliferation are inhibited. Thus, only the organization of the cytoskeleton characteristic of migrating RPE cells that have SFs that interact with the substratum at FCs, is associated with the induction of cell proliferation. PMID- 8714693 TI - Determination versus differentiation and the MyoD family of transcription factors. AB - The myogenic regulatory factors (MRFs) form a family of basic helix-loop-helix transcription factors consisting of Myf-5, MyoD, myogenin, and MRF4. The MRFs play key regulatory roles in the development of skeletal muscle during embryogenesis. Sequence homology, expression patterns, and gene-targeting experiments have revealed a two-tiered subclassification within the MRF family. Myf-5 and MyoD are more homologous to one another than to the others, are expressed in myoblasts before differentiation, and are required for the determination or survival of muscle progenitor cells. By contrast, myogenin and MRF4 are more homologous to one another than to the others and are expressed upon differentiation, and myogenin is required in vivo as a differentiation factor while the role of MRF4 remains unclear. On this basis, MyoD and Myf-5 are classified as primary MRFs, as they are required for the determination of myoblasts, and myogenin and MRF4 are classified as secondary MRFs, as they likely function during terminal differentiation. PMID- 8714694 TI - Schwannomin: new insights into this member of the band 4.1 superfamily. AB - Neurofibromatosis type 2 (NF2) is an autosomal dominant disease characterized by the development of central nervous system tumours. The NF2 gene was recently cloned and found to encode a protein, schwannomin (or merlin), with homology to the band 4.1 superfamily. This superfamily of proteins includes ezrin, moesin, radixin, and talin, as well as several protein tyrosine phosphatases. How does a cytoskeleton-associated protein act as a tumour suppressor? While this fundamental question remains unanswered, recent studies have begun to address key questions regarding the function of schwannomin. In this review, we examine what is known about the band 4.1 superfamily and how this information pertains to schwannomin. In addition, we summarize recent studies of schwannomin itself. PMID- 8714695 TI - An analysis of an axonal gradient of phosphorylated MAP 1B in cultured rat sensory neurons. AB - The present study investigated the cellular distribution of a developmentally regulated phosphorylated form of MAP 1B recognized by monoclonal antibody (mAb) 150 in cultures of dorsal root ganglia. The cell soma and the whole axon, when it first appears, are labelled, but longer axons label with a proximodistal gradient, such that the cell soma and proximal axon become unlabelled, whilst the distal axon and growth cone label strongly. Double-labelling experiments with mAb 150 and a polyclonal antibody (N1-15) that recognizes all forms of MAP 1B demonstrated that MAP 1B is distributed along the entire length of axons with gradients, so the gradient of phosphorylated MAP 1B is not due to a loss or absence of MAP 1B from the proximal axon. The proportion of axons from 20 h cultures that were labelled with a mAb 150 gradient was at least 80% and this proportion was independent of the nerve growth factor concentration of the culture medium. Analysis of axons ranging in length from 100 to 700 microm and labelled with a gradient showed that the unlabelled proximal portions of axons increased in length more slowly than the labelled distal axon. Axons labelled along their entire length accounted for no more than 19% of th axonal population and analysis of these showed them to be frequently < 400 microm long. After simultaneously fixing and detergent-extracting cultures this proportion rose significantly to 93%, suggesting that in the proximal axon the mAb 150 epitope is masked by some factor(s) that is removed by detergent extraction. The possibility that mAb 150 could not access the epitope in the proximal axon was discounted because another IgM, mAb 125, which recognizes a different phosphorylation epitope on MAP 1B, labelled the proximal axon of conventionally fixed cultures. In growth cones of fixed and extracted neurons examined by immunofluorescence, the mAb 150 labelling strongly colocalized to bundled microtubules in the distal axon shaft and the C-domain. In the P-domain, mAb 150 staining was weaker and more widely distributed than the microtubules. Immunogold electron microscopy confirmed that antibody N1-15 and mAb 150 strongly labelled the bundled microtubules in the C-domain and also showed that individual microtubules in the P-domain, some of which lie alongside actin filament bundles of filopodia, were labelled lightly and discontinuously with both antibodies. This suggests that the phosphorylated isoform of MAP 1B recognized by mAb 150 may be microtubules and actin filaments in the P-domain. PMID- 8714696 TI - The role of spinal neurokinin-2 receptors in the processing of nociceptive information from the joint and in the generation and maintenance of inflammation evoked hyperexcitability of dorsal horn neurons in the rat. AB - In spinal cord neurons in anesthetized rats, the role on neurokinin A and neurokinin-2 receptors in the processing of nociceptive information from the knee joint was studied. The specific non-peptide antagonist at the neurokinin-2 receptor, SR48968, its inactive R-enantiomer, SR48965, neurokinin A, substance P and (RS)-alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA), were administered ionophoretically close to neurons with input from the knee joint. SR48968 reduced the effects of exogenous neurokinin A, but not those of exogenous substance P and AMPA, indicating selective blockade of neurokinin-2 receptors. In most neurons with input from the normal knee joint, SR48968 reduced dose dependently the responses to noxious pressure with applied to the knee, and in approximately 50% of the neurons the responses to innocuous pressure. The administration of SR48968 during the induction of an experimental joint inflammation markedly attenuated the development of inflammation-evoked hyperexcitability. In hyperexcitable neurons with input from the inflamed joint, SR48968 reduced the responses to noxious and innocuous pressure. The relative reduction of the responses was more pronounced than in neurons with input from the normal joint. None of the effects of SR48968 was mimicked by SR48965. These data show that neurokinin-2 receptors are involved in the spinal processing of nociceptive information from the normal joint. Furthermore, neurokinin-2 receptors must be coactivated at an early stage of inflammation, to allow the generation of hyperexcitability. Finally, neurokinin-2 receptors are involved in maintenance of hyperexcitability during inflammation. In summary, spinal neurokinin-2 receptors are important in the generation of pain in the normal and inflamed joint. PMID- 8714697 TI - Kinematic analysis of the cat shoulder girdle during treadmill locomotion: an X ray study. AB - A quantitative kinematic analysis of the movements of the shoulder girdle in the three dimensions of space during treadmill locomotion (velocity range 0.33-1.2 m/s) was performed in two cats. Since the movement patterns of the scapula and the humeroscapular joint can only vaguely be estimated through the overlying skin we used implanted metal spheres placed on the scapula in combination with three dimensional pulsed X-ray cinematography (time resolution 20 ms) to reconstruct the excursions of the scapula, the humerus and the elbow and to calculate the respective angular amplitudes and velocities. The movements of the scapula relative to the Th4 spinous process consist of four major components:(i) a monophasic flexion (caudocranial movement of glenoid fossa during swing)/extension (craniocaudal movement of the glenoid fossa during stance) sequence, the fulcrum for which sequence is situated near the vertebral border of the scapula at the medial elongation of the scapular spine; (ii) a vertical monophasic up/down sequence of the fulcrum relative to the trunk, the highest vertical position being reached during mid-stance and the lowest vertical position during mid-swing; (iii) a biphasic abduction/adduction sequence during swing and during stance respectively; and (iv) small rotations along the scapular spine. The trajectory recordings of the scapula indicate that the scapula yields relative to the trunk under the body weight after ground contact. The angular excursions of the humeroscapular joint consist of : (i) a flexion/extension sequence during swing, a yield after ground contact and a final extension at the end of stance; (ii) an adduction and outward rotation during the early swing phase flexion; (iii) an abduction and inward rotation during the late swing phase extension; and (iv) an adduction during the yield with only minor rotations during the whole stance phase. The movement patterns are discussed in view of the muscular synergies necessary to guide the scapula and the humerus during stance and swing, and in relation to the implications for the organization of these patterns in spinal neuronal systems. PMID- 8714698 TI - A p65/p95 neural surface receptor is expressed at the S-G2 phase of the cell cycle and defines distinct populations. AB - A surface receptor complex of Mr approximately 65 000 (p65) and approximately 95 000 (p95) is expressed in cells of the central nervous system of mice. This receptor is recognized by monoclonal antibody 87.92.6 or by reovirus type 3 haemagglutinin as unnatural ligands. The p65/p95 receptor is expressed mostly in neural embryonic precursors undergoing proliferation, especially those in the S G2 phase of the cell cycle. Receptor expression decreases progressively throughout embryogenesis to low but detectable levels in the adult brain. Biochemical characterization revealed that the neural p65/p95 receptor complex is indistinguishable from the p65/p95 receptor expressed in T cells, where receptor ligation leads to a mitogenic block. In neural and lymphoid tissues the p65/p95 receptor (or an associated protein) possesses a tyrosine kinase enzymatic activity. Receptor ligation in neural cells resulted in the rapid tyrosine phosphorylation of cellular proteins which are different from substrates phosphorylated in T cells. Differential substrate coupling to the receptor may account for differences in signal transduction and biology between neural cells and T cells. Further study of this receptor complex may help define important features of neural proliferation, differentiation and survival. PMID- 8714699 TI - Neurotrophin-4/5 maintains the cholinergic phenotype of axotomized septal neurons. AB - We examined the effect of intraseptal or intracerebroventricular (i.c. v.) infusions of NT-4/5 or intraseptal infusions of NGF on the level of immunohistochemical staining of choline acetyltransferase (ChAT)and the low affinity nerve growth factor receptor (LNGFR)in the rat medial septum following unilateral transection of the fimbria. The extent of cell loss in the septum ipsilateral to the lesion, determined by cell counts of ChAT-immunopositive neurons and expressed as a ratio comparing the lesioned to the intact sides, was 0.28 in animals that received an infusion of phosphate-buffered saline (PBS). The ratios were 0.97 and 1.07 in animals that received an infusion of NT-4/5 into the ipsilateral ventricle and septum respectively. Septal infusions of NGF produced a ratio of ChAT-immunopositive cells of 1.03. The ratios of LNGFR-immunopositive neurons increased from 0.50 in PBS-infused animals to 0.79 and 0.83 in animals infused with NT-4/5 via the i.c. v. infusion of NT-4/5 or septal infusion of NT 4/5 or NGF. As determined by immunohistochemical staining, NT-4/5 infused into the lateral ventricle was detected in the periventricular portions of the forebrain ipsilateral to the infusion, while NT-4/5 or NGF infused intraseptally was detected in much of the septum, bilaterally. Furthermore, exogenous NT-4/5 or NGF was detected in numerous neuronal perikarya in the medial septal and diagonal band nuclei. These data demonstrate that, as with NGF, i.c.v. as well as septal infusions of NT-4/5 can maintain the phenotype of basal forebrain cholinergic neurons following axotomy. PMID- 8714701 TI - Less segregated processing of visual information in V2 than in V1 of the monkey visual cortex. AB - To test the possibility of cross-talk between parallel pathways dealing with different aspects of visual information such as orientation, direction of motion and colour in cortical area V2, we quantitatively analysed visual responses of 121 V2 cells recorded from anaesthetized and paralysed macaques and compared them with those of 147 V1 cells. A selectivity index of visual responses was calculated for each neuron, which was then classified as selective or not to a particular attribute of visual stimuli. Twenty-one percent of the V2 neurons had dual selectivity to both colour and direction of stimulus motion (C&D cells). In V1, only 5% of the cells were C&D cells. Thus, the proportion of C&D cells significantly increased from V1 to V2. We also carried out cross-correlation analysis of spike trains recorded simultaneously from pairs of V2 neurons or pairs of V1 neurons. In V2, correlated firings could be observed between cells with completely different optimal orientation, such as orthogonal, while it was never observed in V1. The cross-correlation analysis further indicted that functional interactions in V2 were more widespread than those in V1. These results suggest that neurons which have different functional properties become less segregated, and that functional interactions become more widespread in V2 than in V1. PMID- 8714700 TI - Leukaemia inhibitory factor or related factors promote the differentiation of neuronal and astrocytic precursors within the developing murine spinal cord. AB - Previously we have shown that leukaemia inhibitory factor (LIF) potentiates the development of murine spinal cord neurons in vitro, suggesting that it, or related factors, may play an important regulatory role in neuronal development. We have further investigated this role and show here that the generation of neurons in cultures of embryonic day 10 spinal cord cells is inhibited by antibodies to the beta subunit of the LIF receptor. Since there are more undifferentiated precursors in antibody-treated cultures than in control and LIF treated cultures, it is concluded that the primary action of LIF, or related molecules, is to promote neuronal differentiation, not precursor survival. In addition, the failure of LIF to support neuronal survival in the period immediately following differentiation suggests that the increased numbers of neurons generated with LIF are not attributable to its neurotrophic action. By selecting neuronal precursors on the basis of their inability to express class 1 major histocompatibility complex molecules, it was shown that LIF acted directly upon these cells and not via an intermediary cell. LIF also appears to be involved in regulating the differentiation of astrocytes, since it increases the number of glial fibrillary protein (GFAP)-positive cells present in the cultures and since the spontaneous production of GFAP-positive cells is blocked by antibodies to the LIF beta receptor. These findings suggest that LIF or related factors promote the differentiation of neural precursors in the spinal cord, but that they are not involved in preferentially promoting precursors down a specific differentiation pathway. PMID- 8714702 TI - Tissue-specific and developmental expression of pituitary adenylate cyclase activating polypeptide (PACAP) receptors in rat brain. AB - The two forms of pituitary adenylate cyclase-activating polypeptide, PACAP27,and PACAP38, are novel members of the vasoactive intestinal peptide (VIP)/secretin/glucagon family of peptides. PACAP receptors that are positively coupled to adenylate cyclase and phospholipase C have been recently identified. We examined the expression of PACAP receptors in the rat cortex, hippocampus, cerebellum and hypothalamus during postnatal development. Functional studies revealed PACAP stimulation of cAMP formation in all the brain areas examined and [3H]inositol monophosphate ([3H]InsP) accumulation only in the cerebellum and hypothalamus. Throughout development, the efficacy or PACAP in stimulating cAMP formation slightly increased in the cortex and hypothalamus and decreased in the hippocampus and cerebellum; PACAP stimulation of [3H]InsP formation decreased in the cerebellum and remained steady in the hypothalamus. The effects of PACAP27 and PACAP38 on cAMP levels and inositol phospholipid hydrolysis were dose dependent between 1 and 100 nM. In the same brain areas, treatment with VIP increased cAMP formation at doses greater than 100nM and failed to affect [3H]InsP content, thus suggesting the existence of type-1 PACAP receptors. The reverse transcription polymerase chain reaction (RT-PCR) was used to analyse the mRNA expression of type-1 PACAP receptor splice variants. PACAP receptor gene expression in the central nervous system was regulated in a developmental- and tissue-specific manner. The PACAP-R transcript was detected in all the brain areas examined whereas PACAP-R-hop mRNA ocurred only in the cerebellum and hypothalamus. The different expression profiles and functional properties of PACAP receptors in the developing rat brain suggest an involvement of PACAP in histogenesis, maturation and neurotransmission. PMID- 8714703 TI - The intrinsic optical signal evoked by chiasm stimulation in the rat suprachiasmatic nuclei exhibits GABAergic day-night variation. AB - Infrared light transmittance imaging was used in rat hypothalamic slices to record an intrinsic optical signal (IOS) of the cell ensemble in the suprachiasmatic nuclei (SCN), the locus of the endogenous circadian clock. Upon optic chiasm stimulation, a transient IOS was observed in an area conforming to the known retinohypothalamic tract innervation in the ventral SCN. An increase in extracellular Mg2+ concentration to 10 mM reduced th IOS, suggesting that the elicited IOS is dependent on synaptic transmission. D-2-amino-5 phosphonopentanoic acid and muscimol suppressed the elicited IOS, indicating that NMDA and GABAA receptor-mediated mechanisms were involved in cell ensemble activity reflected in the IOS. The extracellularly recorded spiking of SCN neurons located outside the IOS area remained largely unaffected by the afferent stimulus. Neurons located within the IOS area responded with a depressed electrical discharge, manifesting an inverse relationship between single-unit discharge and the optical measure. The influence of the endogenous circadian rhythm on the elicited IOS was assessed by carrying out daytime-dependent concentration-response experiments. NMDA and non-NMDA receptor specific compounds did not exhibit significant day-night differences, whereas GABA-specific ligands showed a significant day-night variation in activity. The competitive GABAA receptor antagonist bicuculline enhanced the IOS exclusively in the daytime SCN. 5alpha-Pregnane-3alpha,21-diol-20-one (allotetrahydrodeoxycorticosterone), a neuroactive steroid that potentiates GABAergic inhibition, suppressed the IOS in the night-time SCN more than in the daytime SCN. This suggests that in the rat the level of extracellular GABA is higher in night-time SCN compared to the daytime SCN. PMID- 8714704 TI - A single-cell study of the axonal projections arising from the posterior intralaminar thalamic nuclei in the rat. AB - Thalamostriatal projections arising from the posterior intralaminar nuclei (P1; the parafascicular nucleus and the adjacent caudalmost part of the posterior thalamic group) were studied in rats by tracing the axons of small pools of neurons labelled anterogradely with biocytin. Thirteen P1 cells were also stained by juxta cellular application of the tracer. Relay cells of P1 nuclei have a morphology that differs radically from the classical descriptions of the bushy cells which represent the main neuronal type of the sensory thalamic relay nuclei. P1 cells have ovoid or polygonal somata of approximately 20-25 microm, from which emerge four or five thick, long and poorly branched dendrites bearing spines and filamentous appendages; their dendritic domains extend for up to 1.5 mm. Before leaving the nucleus 20% of axons give off collaterals that ramify locally. All axons course through the thalamic reticular nucleus, where they also distribute collaterals, and arborize massively in the striatum and sparsely in the cerebral cortex. At the striatal level four or five collaterals leave the main axon and terminate in patches scattered dorsoventrally within a rostrocaudally oriented slab. As revealed by calbindin D-28k immunohistochemistry, only the matrix compartment receives terminations from P1 axons. The cortical branch form small terminal puffs centred upon layer VI of the motor cortex. Before entering the striatum some axons of the parafascicular nucleus give rise to descending collaterals that arborize in the entopeduncular nucleus, in the subthalamic nucleus and in the vicinity of the red nucleus. Other axons arising from the caudal part of the posterior group send descending branches only to the entopeduncular nucleus. These findings show that P1 cells belong to a distinct category of thalamic relay neurons which, beside their massive projection to the striatum, also distribute collaterals to other components of the basal ganglia. Moreover, these results provide the first direct evidence that virtually all P1 cells project to both striatum and cerebral cortex. Finally, it is proposed on the basis of morphological, histochemical and hodological criteria that the caudal part of the posterior thalamic group in the rat is homologous to the suprageniculate-limitans nuclei of cats and primates. PMID- 8714705 TI - Calpain inhibitors protect against depolarization-induced neurofilament protein loss of septo-hippocampal neurons in culture. AB - We examined the effect of a 6 min depolarization with 60 mM KCl and 1.8, 2.8 or 5.8 mM extracellular CaCl2 on neurofilament proteins of high (NF-H), medium (NF M) and low (NF-L) molecular weight in primary septohippocampal cultures. One day after depolarization, Western blot analyses revealed losses of all three neurofilament proteins. Increasing the extracellular calcium concentration from 1.8 to 5.8 mM CaCl2 in the presence of 60 mM KCl produced increased losses of all three neurofilament proteins to approximately 80% of control values in the absence of cell death. Calcium-dependent losses of the neurofilament proteins correlated with calcium-dependent increases in calpain 1-mediated breakdown products of alpha-spectrin. Calpain inhibitors 1 and 2, applied immediately after depolarization and made available to cultures for 24 h, reduced losses of all three neurofilament proteins to approximately 14% of control values. The protective effects of calpain inhibitors 1 and 2 were influenced by different levels of extracellular calcium. Qualitative immunohistochemical evaluations confirmed semiquantitative Western blot data on neurofilament loss and protection by calpain inhibitors 1 and 2. We propose that brief depolarization causes loss of neurofilament proteins, possibly due to calpain activation. Thus, calpain inhibitors could represent a viable strategy for preserving the cytoskeletal structure of injured neurons. PMID- 8714706 TI - Planning and spatial working memory: a positron emission tomography study in humans. AB - Previous work with both neurosurgical and neurodegenerative patient groups has suggested that high level planning is mediated by neural circuitry which includes both the prefrontal cortex and the striatum. In this study, the functional anatomy of cognitive planning was investigated further, using positron emission tomography (PET). Regional cerebral blood flow (rCBF) was measured in 12 normal volunteers while performing easy and difficult versions of (i) a modified Tower of London planning task; (ii) a mnemonic variant of this task that required short term retention and reproduction of problem solutions; and (iii) a control condition that involved identical visual stimuli and motor responses. Significant increases in rCBF were observed in the left hemisphere, in both the mid dorsolateral frontal cortex and in the head of the caudate nucleus, when the difficult planning task was compared with the control condition. Moreover, subtraction of a simple planning condition from the more difficult one revealed focal increases in rCBF in the caudate nucleus and the thalamus only. During both mnemonic variants of the planning task, changes were also observed in the mid dorsolateral frontal cortex and in more ventral frontopolar regions, bilaterally. When compared directly, the planning and memory conditions differed in terms of these ventral activation foci, but not in the pattern of activation observed in mid-dorsolateral frontal cortex. These findings further implicate frontostriatal circuitry in high-level planning and provide evidence for functionally distinct contributions from ventral and dorsolateral frontal regions to spatial working memory. PMID- 8714707 TI - Chronic alterations in dopaminergic neurotransmission produce a persistent elevation of deltaFosB-like protein(s) in both the rodent and primate striatum. AB - Using an antibody that recognizes the products of all known members of the fos family of immediate early genes, it was demonstrated that destruction of the nigrostriatal pathway by 6-hydroxydopamine (6-OHDA) lesions of the medial forebrain bundle produces a prolonged (>3 months) elevation of Fos-like immunoreactivity in the striatum. Using retrograde tract tracing techniques, we have previously shown that this increase in Fos-like immunoreactivity is located predominantly in striatal neurons that project to the globus pallidus. In the present study, Western blots were performed on nuclear extracts from the intact and denervated striatum of 6-OHDA-lesioned rats to determine the nature of Fos immunoreactive protein(s) responsible for this increase. Approximately 6 weeks after the 6-OHDA lesion, expression of two Fos-related antigens with apparent molecular masses of 43 and 45 kDa was enhanced in the denervated striatum. Chronic haloperidol administration also selectively elevated expression of these Fos-related antigens, suggesting that their induction after dopaminergic denervation is mediated by reduced activation of D2-like dopamine receptors. Western blot immunostaining using an antibody which recognizes the N-terminus of FosB indicated that the 43 and 45 kDa Fos-related antigens induced by dopaminergic denervation and chronic haloperidol administration may be related to a truncated form of FosB known as deltaFosB. Consistent with this proposal, retrograde tracing experiments confirmed that deltaFosB-like immunoreactivity in the deafferented striatum was located predominantly in striatopallidal neurons. Gel shift experiments demonstrated that elevated AP-1 binding activity in denervated striata contained FosB-like protein(s), suggesting that enhanced deltaFosB levels may mediate some of the effects of prolonged dopamine depletion on AP-1-regulated genes in striatopallidal neurons. In contrast, chronic administration of the D1-like receptor agonist CY 208243 to 6-OHDA-lesioned rats dramatically enhanced deltaFosB-like immunoreactivity in striatal neurons projecting to the substantia nigra. Western blot immunostaining revealed that deltaFosB and, to a lesser extent, FosB are elevated by chronic D1-like agonist administration. Both the quantitative reverse transcriptase-polymerase chain reaction and the ribonuclease protection assay demonstrated that deltafosB mRNA levels were substantially enhanced in the denervated striatum by chronic D1-like agonist administration. Lastly, we examined the effects of chronic administration ofD1-like and D2-like dopamine receptor agonists on striatal deltaFosB expression in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) primate model of Parkinson's disease. In monkeys rendered Parkinsonian by MPTP, there was a modest increase in deltaFosB-like protein(s), while the development of dyskinesia produced by chronic D1-like agonist administration was accompanied by large increases in DeltaFosB-like protein(s). In contrast, administration of the long acting D2-like agonist cabergoline, which alleviated Parkinsonian symptoms without producing dyskinesia reduced deltaFosB levels to near normal. Taken together, these results demonstrate that chronic alterations in dopaminergic neurotransmission produce a persistent elevation of deltaFosB-like protein(s) in both the rodent and primate striatum. PMID- 8714708 TI - Effects of repeated sensory stimulation (electro-acupuncture) and physical exercise (running) on open-field behaviour and concentrations of neuropeptides in the hippocampus in WKY and SHR rats. AB - The effects of repeated sensory stimulation (electro-acupuncture) and physical exercise (running) on open-field behaviour and on hippocampal concentrations of neuropeptide Y, neurokinin A, substance P, galanin and vasoactive intestinal peptide (VIP)-like immunoreactivities were studied in WKY (wistar-Kyoto) and SHR (spontaneously hypertensive) rats. Significantly higher concentrations of substance P-like immunoreactivity, neurokinin A-like immunoreactivity and neuropeptide Y-like immunoreactivity were found in the hippocampus immediately after 3 weeks of treatment (electro-acupuncture and running), but not 1 week after the last (tenth) changes in neuropeptide concentrations were similar in the two rat strains. Open-field behaviour was significantly reduced during the treatment period in both strains. There were significant negative correlations between behaviour and neuropeptide concentrations in SHR rats, suggesting interdependency with sympathetic activity. It is proposed that the effects of electro-acupuncture and physical exercise in rats are related to increases in neuropeptide Y, neurokinin A and substance P in the hippocampus. PMID- 8714709 TI - Organization of the visual reticular thalamic nucleus of the rat. AB - The visual sector of the reticular thalamic nucleus has come under some intense scrutiny over recent years, principally because of the key role that the nucleus plays in the processing of visual information. Despite this scrutiny, we know very little of how the connections between the reticular nucleus and the different areas of visual cortex and the different visual dorsal thalamic nuclei are organized. This study examines the patterns of reticular connections with the visual cortex and the dorsal thalamus in the rat, a species where the visual pathways have been well documented. Biotinylated dextran, an anterograde and retrograde tracer, was injected into different visual cortical areas [17; rostral 18a: presumed area AL: (anterolateral); caudal 18a: presumed area LM (lateromedial); rostral 18b: presumed area AM (anteromedial); caudal 18b: presumed area PM (posteromedial)] and into different visual dorsal thalamic nuclei (posterior thalamic, lateral geniculate nuclei), and the patterns of anterograde and retrograde labelling in the reticular nucleus were examined. From the cortical injections, we find that the visual sector of the reticular nucleus is divided into subsectors that each receive an input from a distinct visual cortical area, with little or no overlap. Further, the resulting pattern of cortical terminations in the reticular nucleus reflects largely the patterns of termination in the dorsal thalamus. That is, each cortical area projects to a largely distinct subsector of the reticular nucleus, as it does to a largely distinct dorsal thalamic nucleus. As with each of the visual cortical areas, each of the visual dorsal thalamic (lateral geniculate, lateral posterior, posterior thalamic) nuclei relate to a separate territory of the reticular nucleus, with little or no overlap. Each of these dorsal thalamic territories within the reticular nucleus receives inputs from one or more of the visual cortical areas. For instance, the region to the reticular nucleus that is labelled after an injection into the lateral geniculate nucleus encompasses the reticular regions which receive afferents from cortical areas 17, rostral 18b and caudal 18b. These results suggest that individual cortical areas may influence the activity of different dorsal thalamic nuclei through their reticular connections. PMID- 8714710 TI - Full-lenth cloning, expression and cellular localization of rat plasmolipin mRNA, a proteolipid of PNS and CNS. AB - We have isolated a 1.476 bp cDNA (NTII11) representing a transcript that is differntially expressed during sciatic nerve development and regeneration in the rat. Nucleotide sequence comparison indicates partial identity with a recently isolated plasmolipin cDNA. However, our clone extends the published sequence by 234 bp at the 5' end and predicts a protein that contains an additional 25 amino acids at th N-terminus. The open reading frame of th NTII11 transcript encodes a 19.4 kDa protein with four putative transmembrane domains. Northern blot analyses revealed a tissue-specific expression was confirmed by in situ hybridization, and cellular localization of plasmolipin mRNA was demonstrated in Schwann cells of the sciatic nerve and in glial cells of myelinated brain structures. The steady state levels of plasmolipin mRNA were markedly altered (i) during development of sciatic nerve and brain. (ii) after sciatic nerve injury, and (ii) in cured Schwann cells maintained under different conditions of cell growth and arrest. Our data indicate a function of plasmolipin during myelination in the central as well as in the peripheral nervous system. PMID- 8714711 TI - Acquisition of conditioned inhibition in rats is impaired by ablation of serotoninergic pathways. AB - The effects of chronically ablating the serotoninergic inputs to various regions of the rat brain on the ability to solve a feature-negative discrimination was measured. After intracerebroventricular administration of the specific neurotoxin 5,7-dihydroxytryptamine, the rats exhibited an impaired capacity to solve such a discrimination, irrespective of whether auditory or visual stimuli were used. Further behavioural analysis revealed that this effect was not due to a reduced capacity to form excitatory associations, since both groups responded equally to reinforced stimuli. By contrast, the lesion more likely resulted in a failure to endow the non-reinforced stimuli with inhibitory properties. This suggestion was supported by the observation that, in a retardation test, the conditioned inhibitor aroused less inhibition in the lesioned group than in vehicle-injected controls. Furthermore, the conditioned inhibitor failed to pass a summation test in lesioned animals, again indicating that their hampered ability to master the discrimination was the result of an impairment in the formation of inhibitory associations. It is concluded that destruction of central 5-hydroxytryptamine containing pathways impairs the functioning of brain areas underlying inhibitory associative learning. PMID- 8714712 TI - Macrophage inflammatory protein-1beta (MIP-1beta) produced endogenously in brain during E. coli fever in rats. AB - Macrophage inflammatory protein-1 (MIP-1) evokes an intense fever, independent of a prostaglandin mechanism, and is now thought to play an important role in the defence response to bacterial pyrogens. The purpose of this study was 2-fold: (i) to determine whether the potent doublet of this cytokine, MIP-1beta, is actually produced in the brain in response to a pyrogenic dose of a lipopolysaccharide of Escherichia coli and (ii) to determine the anatomical site of synthesis of this cytokine in the brain. Following the intense fever produced by intraperitoneal administration of lipopolysaccharide in the unrestrained rat, MIP-1beta immunoreactivity was identified post mortem in two regions of the brain implicated in fever: the organum vasculosum laminae terminalis (OVLT) and the anterior hypothalamic, preoptic area (AH/POA). Microinjection of goat anti-mouse MIP-1beta antibody (anti-MIP-1beta) directly int the AH/POA markedly suppressed fever in rats in response to lipopolysaccharide. Further anti-MIP-1beta administered 180 min after the injection of lipopolysaccharide acted as an antipyretic and reversed the fever induced by the endotoxin. anti-MIP-1beta or control immunoglobulin G antibody microinjected into the hypothalamus immediately before the intraperitoneal injection of the control saline did not alter the temperature of the rats. Taken together, the present results demonstrate that MIP 1beta is produced in the brain in response to a bacterial endotoxin. These observations, in the light of earlier data on fever induced by MIP-1beta, further support the hypothesis that endogenously synthesized MIP-1beta acts as an intermediary factor in the evocation of fever by acting on the thermosensitive cells of the brain. PMID- 8714714 TI - Bionic artificial kidney. PMID- 8714713 TI - Expression of glycine receptor alpha subunits and gephyrin in cultured spinal neurons. AB - The inhibitory glycine receptor is a pentameric membrane protein composed of alpha and beta subunits. In the postsynaptic membrane, the glycine receptor and the copurifying peripheral membrane protein gephyrin are clustered underneath glycine-releasing nerve terminals. Here, we describe the expression of gephyrin and the neonatal and adult glycine receptor alpha subunit isoforms alpha1 and alpha2 during in vitro differentiation of rat spinal neurons. Analysis by immunoassays and the reverse transcriptase-polymerase chain reaction showed that gephyrin and alpha subunit mRNA and protein levels exhibited a marked increase from 1 to 5 days in vitro, i.e. prior to the formation of functional synaptic contacts. Using confocal and standard immunofluorescence, we determined the number of immunoreactive cells and the cellular localization of the alpha subunits and gephyrin. At 3 days in vitro, glycine receptor immunoreactivity revealed by the monoclonal antibody mAb4a was found in < 10% of cells and was mainly localized intracellularly; in contrast, gephyrin was detected in in vitro, gephyrin was essentially localized at the neuronal surface. At this stage, the number of glycine receptor-positive cells approached that of gephyrin-containing neurons (50%), and glycine receptor antigen was found both intracellularly and at the periphery of the cells. The antibody mAb2b, which binds exclusively to the alpha1 subunit, revealed aggregates at the surface of a few neurons. At 10 days in vitro, glycine receptor and gephyrin staining was localized in clusters at the periphery of the soma and the neurites. This quantitative analysis corroborates temporal differences in the cellular distribution of gephyrin and glycine receptor alpha subunits, the former being accumulated first at the neuronal surface. PMID- 8714715 TI - A variety of clinical applicabilities of immobilized dextran sulphate as lipoprotein adsorbent and avoidance of anaphylactoid (anion-blood contact) reaction in its use. AB - Dyslipidemia including hyper-LDL(low density lipoprotein) cholesterol which is very often refractory to dietary/medical treatments is known to be a risk factor of many arteriosclerotic lesions. An extracorporeal procedure of plasma adsorption, LDL adsorption, utilizing dextran sulphate as a ligand immobilized on cellulose gel beads has been clinically applied in a variety of dyslipidemic conditions as listed below. Its usefulness in secure reduction of the serum LDL level and consequent symptomatic improvements has been confirmed. Familial hypercholesterolemia(FH): A regular repetition of the LDL adsorption ameliorates hyper-LDL cholesterolemia as resulting in regression of the multiple stenoses in the coronary arteries. Focal glomerulosclerosis(FGS): A seesion of the LDL adsorption improves kidney function and reduces a urinary protein excretion in FGS patients with dyslipidemia. Arteriosclerosis obliterans(ASO): More than 60 ASO patients with dyslipidemia have been treated by the LDL adsorption in our center. In over 80% of the patients, marked improvement in clinical symptoms such as leg pain/intermittent claudication has been brought out. Hemodialysis-relevant dyslipidemia(HDDL): HDDL which develops in the long-term HD patients has been treated. Transplantation-relevant dyslipidemia(TXDL): TXDL with deterioration of the transplanted kideny function has been treated by the LDL adsorption. Kidney function improves. Anaphylactoid reaction which is tentatively explained as a result of release of bradykinin in contact of blood with polyanionic material of the adsorbent, dextran sulphate, develops, in particular, while an angiotensin converting enzyme inhibitor is administered as a depressant. However, it can be avoided in a use of nafmostat mesilate, a protease inhibitor, as an anticoagulant. PMID- 8714716 TI - Systemic responses to SFHS-infusion in hemorrhaged dogs. AB - Anesthetized mongrel (weight range: 16-27 Kg) dogs were prepared for monitoring hemodynamics, blood flow distribution, plasma colloid osmotic pressure and renal functional parameters at various intervals. Removal of 35 ml/Kg blood resulted in marked drop and only partial spontaneous recovery in systemic and pulmonary arterial pressures, cardiac output and organ blood flows (> 50% flow-decrements occurred in kidney, spleen, heart, gut and pancreas); plasma colloid osmotic pressure as well as urine output and creatinine clearance also fell. Group I (n = 6) of dogs was transfused after 45 minutes of hypovolemia with their own anticoagulated blood, while Group II (n = 6) received an equal volume of unmodified 6% stromafree hemoglobin solution (SFHS). Comparison of the two groups' responses to resuscitation yielded some differences. There was a significant overshoot (30 mmHg) in systemic arterial blood pressure accompanied by bradycardia in Group II only. Cardiac output recovered in both groups but was less well sustained in Group II. Cerebral blood flow rose higher and hepatic arterial flow-increment was less in Group II than in Group I; the responses to resuscitation in the other organs were comparable. Colloid osmotic pressure decreased in Group I whereas it rose immediately after resuscitation in Group II, declining thereafter with a converging trend and 30 minutes thereafter the differences were not significant between the groups. Urine excretion and creatinine clearance recovered to comparable extents in both groups, but N-acetyl beta-D-glucosaminidase (N.A.G.) excretion rose over 10-fold higher in Group II than in Group I. These experiments have defined the response of bled animals to resuscitation with unmodified, unpurified SFHS, when compared to resuscitation with whole blood, showing a less well sustained but adequate hemodynamic and renal functional recovery while revealing indications of early renal tubular cellular injury, providing baseline comparison for testing highly purified and modified hemoglobin solutions. PMID- 8714717 TI - Alteration of cerebral microcirculation by hemodilution with hemosome in awake rats. AB - Our study showed that hemodilution with modified fluid gelatin resulted in an increase in local cerebral blood flow (LCBF), but no change at all in local cerebral oxygen delivery (LCOD) in rats. Hemosome, a lecithin encapsulated hemoglobin having the oxygen-carrying capacity, was developed to improve LCOD by hemodilution. Therefore, we have hypothesized that LCBF & LCOD would be increased by hemodilution with hemosome. To test this hypothesis, adult male Sprague-Dawley rats weighing approximately 350g were used and divided into the hemodilution and the control groups. Hemosome was made from pig red blood cells and lecithin. It's mean diameter was approximately 0.3 um and hemoglobin concentration was approximately 4g/dl. Isovolemic hemodilution, which lowered the systemic hematocrit from approximately 50% to approximately 30%, was achieved by rapidly replacing blood with the same volume of hemosome. Ten min later, LCBF in 14 brain structures were measured using the 14C-iodoantipyrine technique. Our results showed that LCBF of the control group ranged from 115 +/- 11 ml/100g/min in the medulla to 260 +/- 31 ml/100g/min in the occipital cortex. LCBFs were generally higher (p < 0.05, MANOVA) by 16% in the hemodilution group than in the control group. However LCODs were generally decreased (p < 0.05, MANOVA) by 18% in the hemodilution group than in the control. In conclusion, hemodilution with hemosome indeed improves LCBF but lowers LCOD in awake rats. PMID- 8714718 TI - Study on in vitro stability of polymerized liposomes. AB - We studied the stability of liposomes composed of polymerized phospholipid, 1,2 bis-(octadeca-2,4-dienoyl)-sn-glycero-3-phosphocholine (DENPC), to pH changes and the presence of bile salts, monitoring the leakage of 3H-sucrose entrapped in liposomes. The stability of polymerized DENPC-liposomes to acidic media (pH 3.3 and 2), and the presence of bile salts (20 mM), was much higher than that of nonpolymerized DENPC-liposomes and liposomes composed of conventional phospholipids. The stability of polymerized DENPC-liposomes was increased in the presence of carboxymethyl chitin (CM-chitin). PMID- 8714719 TI - Transplantation of human endothelial cell monolayer on artificial vascular prosthesis: the effect of growth-support surface chemistry, cell seeding density, ECM protein coating, and growth factors. AB - The failure rates of synthetic vascular grafts, when placed in low blood flow environments in humans, are not acceptable. Thus, endothelial cell (EC) seeding technology of vascular grafts was developed to prepare prostheses lined with a human monolayer expressing optimal thromboresistant properties. In a clinical setting, endothelialization of a graft can be achieved using higher cell seeding densities, or by creating a surface on which EC can adhere and grow to confluence. But, human endothelial cells show little or no proliferation on the currently available graft materials. In this study, surface modification of PTFE and ePTFE by ammonia plasma treatment was carried out to enhance its interactions with ECM protein, EC growth factors, and with EC harvested from human umbilical vein (HUVEC), and from human saphenous veins (HSVEC). Our data shows that various vascular graft materials generated from ammonia plasma treated PTFE and ePTFE exhibited statistically significant improvements in HUVEC and HSVEC growth when compared to their respective controls (p values < 0.001). Growth of HSVEC on ammonia plasma treated ePTFE without ECM protein coating was also found to be statistically significant in comparison to that on fibronectin coated ePTFE (p < 0.001). The final HSVEC cell densities found on various ePTFE surfaces prepared from ammonia plasma treated ePTFE, suggests that transplantation of HSVEC monolayers on vascular prostheses can be established within clinically relevant times. Ammonia plasma treatment process provides an unique opportunity to surface modify prosthetic materials of various construct to transplant mammalian cells including those that have undergone ex vivo gene transfer, and to deliver angiogenic molecules to a target area for tissue development. PMID- 8714720 TI - Antibodies released from immunoadsorbents: effect of support, activation and elution conditions. AB - Immunoadsorption is an application of affinity chromatography, as a therapeutic method to specifically deplete biological fluids such as blood plasma from proteins in excess, or to extract a biomolecule from a complex mixture. However, the leakage of small amounts of antibodies covalently immobilized on the support hampers the practical use of this method. In fact, these released antibodies contaminate the purified proteins or depleted media and, when they are of animal nature, they may lead to immunization of patients, or cause an anaphylactic shock when a clinical use is concerned. It is therefore of prime importance that the immunoadsorbents exhibit a satisfactory stability over the whole range of chemical and biochemical conditions involved during their clinical handling. To determine optimal conditions for the preparation of stable immunoadsorbents designed to remove selectively Low Density Lipoproteins (LDLs) from the plasma of patients affected by familial hypercholesterolemia, various immunoadsorbents were prepared by covalent immobilization of goat anti-apolipoprotein B polyclonal antibodies on different supports (Sepharose CL-4B, Sepharose 6 Fast Flow, Spherodex and Fractogel) previously activated by various chemical reagents (cyanogen bromide, divinyl sulphone, tresyl chloride and trichloro-s-triazine). Their adsorption capacity, specificity, stability and the amount of immobilized antibodies were compared in terms of the activation method and the support used. It turns out that the immunoadsorbents prepared with Sepharose 6 Fast Flow lead to optimal yield of coupling, adsorption capacity, and an excellent stability at neutral pH. TC-activated-Fractogel turns out as well to afford an excellent coupling yield, a good adsorption capacity and an optimal stability in the whole pH range tested. PMID- 8714721 TI - -60-th anniversary of the Lodz section of the Polish Neurological Society-. PMID- 8714722 TI - -The evaluation of the effectiveness and tolerance of sumatriptan in abortive treatment of migraine attacks (open multicenter trial). AB - The authors review data on sumatriptan obtained in the results of their own studies performed on 78 patients in 10 Polish neurological centres. In all 154 migraine attacks were treated with 6 mg of subcutaneous sumatriptan. Significant decrease of pain intensity was achieved in 30% of cases after 15 min and 60%- after 30 min. According to the patient's own opinion, 62% of them evaluated the drug as very efficient, 23% of patients--as efficient. Adverse effects were reported by 11 patients (15%), but they were mild and transient. Headache recurred in 16% of patients. PMID- 8714723 TI - [Current views on pathophysiology of migraine: Part I. Genetics of migraine. Genesis of the vascular theory]. AB - The studies on the pathogenesis of migraine have developed into a vast scientific movement in last years. The author discusses throughly the results of these studies. In the first part of the paper the genesis and development of the vascular theory of migraine pathogenesis is presented. PMID- 8714724 TI - [Current views on pathophysiology of migraine. Part II: Further development and current status of the vascular theory. Migraine and allergy]. AB - In this part further arguments advocating the vascular theory are discussed, among others: the influence of drugs (ergotamine, sumatriptan), so called "migrainous stroke", and the results of the last neuroimaging studies on the migraine. The authors state in summary that vascular disturbances do take place in migraine attack, but they should be regarded in connection with biochemical and central neuronal disorders. In the end the authors discusse the current views on the impact of allergy in the pathophysiology of migraine. PMID- 8714725 TI - [Current views on pathophysiology of migraine. Part III: Biochemical hypothesis. The role of blood platelets]. AB - Systematic studies on the biochemistry of migraine were first undertaken in early 1960s; they initially concerned serotonin metabolism. The studies were next extended, covering catecholamines, histamine, free fatty acids, selected peptides and hormones. The results were mostly disappointing and such studies were mostly given up. In the last years we have been witnessing the come back of the study on the role of serotonin in migraine as the neurotransmitter involved in the antinociception and vascular innervation. In the 1970s a great deal of studies on blood platelets as serotonin reservoir were undertaken in migraine patients. PMID- 8714726 TI - [Current views on pathophysiology of migraine. Part IV: autonomic disturbances. Central neural hypothesis. Conclusions]. AB - The last part of the article concerns the role of the disturbances of autonomic system and central neural hypothesis of migraine pathogenesis. The author presents the results of studies on pupillometry and autonomic heart regulation, that seem to indicate hypofunction of the autonomic system in migraine. The views on "neurogenic inflammation", as the basic pathologic phenomenon in migraine attack are also discussed, as well as the hypothesis of cortical spreading depression. In the conclusions the author find that neither theory can convincingly explain the pathogenesis of migraine. The updated studies seem to indicate both vascular, autonomic and central neural factors in, possibly multifactorial, pathogenesis of migraine. PMID- 8714727 TI - [Automatic regulation of sinus rhythm in patients with migraine]. AB - The clinical symptoms of migraine point to autonomic disturbances, especially to disrupted regulation of the circulatory system and autonomic balance. Searching for more accurate autonomic system studies we turned to the spectral analysis of cardiac rhythm changes, that allows to estimate the autonomic balance in sinus node. 44 patients with migraine were studied and the results were compared with those obtained in 74 healthy subjects. The reduced influence of both sympathetic and parasympathetic part of autonomic system on the sinus rhythm in migraine patients was found. The autonomic balance is shifted to the parasympathetic innervation side in patients with migraine. PMID- 8714728 TI - [Ultrasound studies of blood flow velocity in main cranial arteries in idiopathic headaches]. AB - The published results of transcranial Doppler (TCD) studies of main cerebral arteries, performed in the patients with idiopathic headaches, especially with migraine, are controversial. Some authors could not find any changes, others observed increased blood flow. These results were, however, based on single examination, and the comparison on mean velocities calculated for the whole groups. In the present study the authors decided to perform a series of TCD studies in each patient and to analyse the individual cases. It was found that blood flow velocity in the main cerebral arteries in chronic tension-type headache (8 cases) and cluster headache (7 cases) showed normal values, established for 35 healthy subjects. In migraine without aura (20 cases) and with aura (11 cases) a considerable increase (above 30%) in cerebral blood flow was found in some studies. In around 50% of the cases increased blood flow in anterior and middle cerebral arteries was found unilaterally. The authors believe that the above findings might be specific for migraine and reflect transient vasoconstriction of the vessels that, on the other head, may be the premonitory laboratory sign of the attack. PMID- 8714729 TI - [Alternating hemiplegia as a migraine equivalent in a small child]. AB - The authors present the diagnostic criteria, clinical course and additional examinations in alternating hemiplegia (ah). According to International Headache Classification ah belongs to the syndromes which precede or accompany migraine. We give a description of the case which is the first published in Polish literature and discuss the etiology supporting the connection of ah with migraine. PMID- 8714730 TI - [The comparison of sodium valproate and ergotamine titrate plus caffeine in the abortive treatment of migraine attacks]. AB - The contemporary measures against migraine attacks are not fully satisfying, thus the migraine episode is still a challenge in treatment. In 1993 Hering and Steiner proposed valproic acid for the interruption of migraine attacks. The authors present the results of their own study comparing the effects of sodium valproate (SV) and ergotamine plus caffeine (Coffecorn) in this respect. 82 attacks in 20 patients were treated altogether. The effectiveness of SV was statistically not different from that of ergotamine--the drug recognized formerly as the first choice one in the abortion of migraine attacks. PMID- 8714731 TI - [Sodium valproate versus propranolol in the prophylactic treatment of migraine]. AB - In the recent years sodium valproate (SV) has been proposed as a prophylactic drug in migraine. Several reports documented a positive effect of SV in migrainous patients. The authors present the results of the open study in 35 women with migraine without aura treated with the daily dose of 1000-1500 mg of SV, during 10 weeks. The results were compared with the effect of propranolol administered to the same patients, in daily dose of 120-160 mg during 10 weeks. The effects were similar: in both methods more than 50% reduction of frequency and severity of attacks was obtained. The side effects were generally mild; in no case the treatment was stopped. The authors conclude that in the future SV might be administered in migraine prophylaxis as the first choice drug. PMID- 8714732 TI - [Results of surgical treatment of patients after spinal trauma with partial spinal cord damage]. AB - Treatment of spinal injuries is a special line in the Dept. of Neurosurgery of Military Academy in Lodz. The authors reviewed the results of surgical treatment of cervical spine injuries in the Dept. between 1982-1987. Follow-up was performed in a group of 49 cases, who vosited the Dept. between 5 to 1 year after surgery. This group of patients underwent detailed clinical analysis in terms of long-term results of treatment. Radiological results were assessed as well. The authors analyzed the relationship between timing of surgical decompression of the spinal cord and clinical improvement of neurological status. The concluded that early decompression in case of incomplete spinal cord injury resulted in better neurological outcome. PMID- 8714733 TI - [Statistics of brain metastases]. AB - Frequency and epidemiology of brain metastases are discussed on the basis of autopsy and clinical reports from different countries. Polish bibliography on this subject is also reviewed. Combining data on the frequency of brain metastases from a large autopsy series and the number of cancer deaths from Polish Cancer Registry, the table was built showing the number of probable patients with brain metastases in Poland. The author suggests the necessity of further research on the frequency and results of treatment of brain metastases in Poland. PMID- 8714735 TI - [Comments on satellite symposium on the treatment of patients with multiple sclerosis with interferon-beta-1b, Munich (Germany), June 19, 1995]. PMID- 8714734 TI - [Automatic analysis of frequency spectrum in localization of topography of epileptic discharges at early stages of the disease]. AB - The authors present the value of frequency spectrum analysis in the method of cerebral mapping for the diagnosis and localization of epileptic focus. The patient aged 17 years showed atypical clinical symptoms. The study made possible confirmation of epilepsy, demonstration of two independent epileptogenic foci, and correlation of the atypical clinical syndrome with the location of the foci. The results are presented on topographic maps of the brain. PMID- 8714736 TI - Imaging of single antigens, antibodies, and specific immunocomplex formation by scanning force microscopy. AB - The most sensitive analytical techniques available today for detecting immuno assay complexes are radio or enzyme immuno analytical techniques, by which quantities of 10(7)-10(8) analyte molecules can be detected. With the introduction of scanning force microscopy, a new method for detecting biological processes became available. Here, we examine the feasibility of using scanning force microscopy as a biosensitive tool. We demonstrate that single or multiple rabbit anti-human serum albumin molecules form complexes with preadsorbed single human serum albumin molecules on mica. However, no interaction is observed between human immunoglobulin G molecules and preadsorbed single albumin molecules; only separate antigens and antibodies are observed at random positions on the mica. This shows the ability of scanning force microscopy to act as a biosensor for detection of immunocomplexes, and to act as a very powerful tool to study molecule-surface interactions in general. PMID- 8714737 TI - Imaging of the cytoplasmic leaflet of the plasma membrane by atomic force microscopy. AB - The cytoplasmic face of ventral cell membranes of Madin-Darby canine kidney (MDCK) cells grown on glass coverslips was imaged by atomic force microscopy (AFM) in air and under aqueous medium, in "contact" mode. Micrometer range scans on air-dried samples revealed a heterogeneous structure with some filaments, likely corresponding to actin filaments that abut the inner leaflet of the membrane, and a few semi-organized lattice structures that might correspond to clathrin lattices. Experiments in phosphate-buffered saline confirmed the heterogeneity of the inner membrane surface with the presence of large (> 100 nm) globular structures emerging from the surface. Using sub-micrometer scan ranges, protruding particles, that occupy most of the membrane surface, were imaged in liquid medium and in air. These particles, 8 to 40 nm x-y size, were still present following ethanol dehydration which extracts a large fraction of membrane lipids, indicating their proteic nature. Due, at least partly, to the presence of some peripheral proteins, high magnification images of the inner membrane surface were heterogeneous with regard to particle distribution. These data compare with those previously reported for the external membrane leaflet at the surface of living MDCK cells. They show that details of the cytosolic membrane surface can be resolved by AFM. Finally, the images support the view of a plasma membrane organization where proteins come into close proximity. PMID- 8714738 TI - High resolution electron microscopy of the interface between dental calculus and denture resin. AB - Dental calculus may grow on the denture surface. In order to demonstrate the mechanism of deposition, the interface between calculus and denture resin was investigated using a high resolution electron microscope. Ultrathin sections were also used for electron diffraction of selected areas to reveal any mineral phase. The mineral layers without mineralized bacteria adjacent to the denture surface revealed a marked variation in thickness and crystal shape. Three types of crystal shape were observed at the junction: needle-like, rod-like and plate-like crystals. High resolution electron microscopy (HREM) showed that both rod-like and plate-like crystals were an aggregation of fine crystallites. The lattice fringes of the fine crystallites were observed among the near atomic structures of resin polymer at the interface in all three types of crystals. The electron diffraction patterns of selected areas revealed that needle-like and rod-like crystals were composed of hydroxyapatite (OH-AP), while plate-like crystals were composed of a mixture of OH-AP and whitlockite. These findings indicate that, after saliva penetrates through the acrylic resin, calcium and phosphate ions in the saliva are trapped in the molecular chains of the resin polymer, while the local ion concentration then increases to reach supersaturation, whereas a spontaneous precipitation would occur at the superficial layer of the denture resin. Furthermore, a thin intermediate layer of crystallites might be indispensable for the scaffolding process in the calculus formation on the denture surface. PMID- 8714739 TI - Mammalian enamel prism patterns and enamel deposition rates. AB - Enamel prism patterns and enamel deposition rates were compared for specimens representing six mammalian orders. Enamel samples were characterized by either pattern 1 or pattern 3 prisms. Each prism pattern category contained prisms from at least two mammalian orders. Enamel deposition rate was estimated for each sample by measuring prism cross striation repeat intervals. Statistical analysis of cross striation repeat intervals illustrates significant differences in deposition rate between prism patterns 1 and 3. No statistically significant differences were found in deposition rate between the higher-level taxa represented within each prism pattern category. That enamel deposition rate is not taxonspecific reinforces the close association between deposition rate and prism morphology. In accord with previous studies, pattern 1 enamel is deposited more slowly than is pattern 3 enamel. Correlation analyses illustrated a lack of association between enamel deposition rate and body mass, tooth size, and estimated ameloblast size. Evidence that enamel deposition rate is associated with enamel prism morphology, coupled with evidence that deposition rate is not correlated with size parameters, points to developmental homology (i.e., homogeneous deposition rate) within each prism pattern. PMID- 8714740 TI - Scanning electron microscope study of the healing molar tooth extraction socket in the rat. AB - Healing molar tooth extraction wounds in rats were examined by scanning electron microscopy from 15 minutes to 40 days following tooth removal. The wound epithelium, which was derived mainly from the gingiva but also from the cheek and hard palate, migrated beneath the superficial socket contents. The contents were lost between 5 to 11 days, thus leaving a central epithelial-lined depression. This decreased in width with time as the level of the wound epithelium approached that of the hard palate but was still present at 40 days. Between 5 and 7 days, the wound epithelium became more regular. However, from 11 days on, it became more irregular with increasing numbers of saucer-shaped depressions, circular defects and circular whorls of epithelial cells. The surface structure of the epithelial cells changed as it migrated and matured. The initially plump, then flattened cells mostly had smooth areas along with variable numbers of irregular microridges and microvilli, although cells derived from the cheek had only smooth surfaces. With further maturation, all cells developed a regular honeycomb surface pattern of interconnecting microridges similar to that on the hard palate. Why the wound epithelium became more uneven after 11 days is not known. PMID- 8714741 TI - Revascularization of an excisional wound in gingiva and oral mucosa. A scanning electron microscopic study using corrosion casts in rats. AB - The purpose of this study was to examine microvascular regeneration associated with gingival wound healing. A full-thickness piece of gingiva and oral mucosa was excised along the palatal aspect of the right maxillary first and second molars in 20 young Wistar rats. The contralateral side served as unoperated control. After 2, 4, 7, 10 or 20 days of healing, microvascular corrosion casts were produced and examined by scanning electron microscopy. At 2 days, vessels surrounding the wound were dilated and impressions representing sites of leukocyte margination were prominent in the walls of venules. Capillary buds were emerging from venules and capillaries. At 4 days, the vessel buds had lengthened and connected in pairs to produce capillary loops. At 7 days, new vessels extended deeply into the wound space, mainly from the medical side, in a palisade like pattern. At 10 days, the denuded bone surface was still not completely revascularized and Volkman's canals opening to the wound area were empty. At 20 days, the bone surface was covered by large, irregular vessels which originated mainly from the palatal mucosa. The periodontal ligament was less important in the tissue repair process, while the bony vasculature contributed little or not at all to revascularization of the healing gingiva and palatal mucosa. PMID- 8714742 TI - The effect of ArF-excimer laser irradiation of the human enamel surface on the bond strength of orthodontic appliances. AB - This study investigated enamel laser conditioning as an alternative to acid etching in bracket therapy. In preliminary experiments optimal laser parameters for achieving a bond strength of 6-10 N/mm2 were defined. Enamel surface morphology was assessed and the ablation depth was measured on serial enamel sections. Thirty human molars were exposed to 193 nm ArF-excimer laser radiation (energy density: 260 mJ/cm2) by single pulse application of 23 nanoseconds. Thirty molars were etched with phosphoric acid (37%) for 60 seconds. The brackets from the treated molars and 30 untreated molars were debonded vertically for tensile bond strength measurement. Roughened enamel surfaces were attained by 450 and 900 laser pulses with a mean ablation depth of 10.13 +/- 4.84 microns. After 1-10 laser pulses, the enamel surface appeared intact. The tensile bond strength was 6.63 +/- 2.18 N/mm2 in the laser-treated group (1 pulse), 8.75 +/- 3.61 N/mm2 in the acid-etched group, and 4.61 +/- 3.15 N/mm2 in the untreated group. We conclude a laser-selective ablation of the membranous enamel pellicle. Since the irradiated area can be adapted to bracket base and the enamel surface remains morphologically intact, pulsed ArF-excimer laser treatment seems to be superior to the acid etching technique. PMID- 8714743 TI - Low concentrations of mercury induce changes in ion composition of cultured myoblasts. AB - The effects of low concentrations (1 pM, 1 nM, 1 microM) of mercuric chloride on ion distribution in cultured myoblasts were analysed by energy dispersive X-ray microanalysis. An increase in intracellular sodium concentration was observed five minutes after addition of HgCl2 to the culture medium. This increase was dose dependent and accompanied by a transient decrease in potassium concentration. Exposure to 1 nM and 1 microM HgCl2 led to a two-fold increase in the cytoplasmic chlorine concentration. The higher HgCl2 concentration (1 microM) induced morphological alterations in the form of cell membrane blebs, perforations and shrinkage or flattening of the myoblasts. It was concluded that even low concentrations of mercuric chloride cause elemental and morphological changes in cultured myoblasts, which may reflect effects of the metal on membrane permeability. PMID- 8714744 TI - Heterogeneous distribution of ecto-Ca-ATPases in primary cultures of human adenohypophyseal cells. AB - The aim of this study was to investigate the localization of ecto-Ca-adenosine triphosphatase (ecto-Ca-ATPase) in different parenchymal cells of the human pituitary in tissue culture. The distribution of ecto-ATPases on the surface membrane of a particular parenchymal cell varied with the type of cells in contact with this parenchymal cell; the membrane portions immediately exposed to the medium showed low if any ecto-ATPase activity. These results suggest that ecto-Ca-ATPases of the parenchymal cells may be involved in cell adhesion processes and may be of crucial importance in the organization (in vivo) and reorganization (in vitro) of human adenohypophyseal tissue. PMID- 8714745 TI - The morphological effects of two antimicrobial peptides, hecate-1 and melittin, on Escherichia coli. AB - The effects of the 26 amino acid, cationic, amphipathic, antibacterial peptide melittin and hecate-1, a 23 amino acid analog of it, on the gram negative bacterium Escherichia coli were investigated using scanning electron microscopy (SEM), transmission electron microscopy (TEM), and freeze-fracture. Both peptides killed virtually all bacteria at the peptide concentration and cell density used. TEM and SEM revealed aggregates of bacteria entangled with material extruded from the bacterial surfaces. SEM revealed irregular bacterial surfaces with bleb-like projections. TEM and freeze-fracture indicate that the bacterial inner and outer membranes, as well as the peptidoglycan layer between, were extensively damaged. The cytoplasmic contents of the cells, however, did not appear radically disturbed, providing little evidence for osmotically induced cytolysis. PMID- 8714746 TI - Apoptosis induced by microtubule disrupting drugs in normal murine thymocytes in vitro. AB - Disruption of cytoplasmic and spindle microtubules by colchicine or nocodazole increases mitotic index, but it also enhances apoptosis in isolated mouse thymocytes; the apoptotic index exceeds 20% after 4 hours of incubation with either drug (5% in controls). Apoptosis was confirmed by DNA fragmentation, and was blocked by calcium chelators and inhibitors of protein synthesis. The apoptotic effect of microtubule disrupting drugs (MDD) was directed to interphase thymocytes and was independent on MDD action on mitotic cells. However, cell death of mitotically arrested cells showed ultrastructural changes similar in many aspects to apoptosis. PMID- 8714747 TI - Cell death in the embryonic developing limb. AB - In amniote vertebrates, the development of form and structure of the limb bud is accompanied by precise patterns of massive mesodermal cell death with morphological features of apoptosis. These areas of cell death appear to eliminate undifferentiated cells which are required only for a limited time period of limb development. Predictable skeletal and morphological anomalies of the limb occur when the pattern of cell death is modified in mutant species or under experimental conditions. Most evidence points to the occurrence of local triggering mechanisms to account for the establishment of the areas of cell death and the subsequent activation of cell death genes. Modifications of the extracellular matrix and diminution in the contribution of growth factors by neighbouring tissues appear as the most likely potential candidates for triggering the cell death program. Information on the genetical basis of cell death in the developing limb is very scarce. Among the increasing number of cell death genes identified in other cell death systems, such as p-53 and the ced 3/ICE and ced-9/ bcl-2 gene families, only bcl-2 has been studied in detail during limb development and yet, the information obtained is contradictory. Bcl-2 is not expressed in the areas of cell death of the developing limb, but normal limbs develop in mice with disruption of the bcl-2 gene. Obviously, the clarification of the role of the cell death genes constitute a major task in future studies of cell death in the developing limb. PMID- 8714748 TI - Radiation nephropathy: a review. AB - The marked radiosensitivity of renal tissue represents a limitation on the total radiotherapeutic dose that safely can be applied to treatment volumes that include the kidneys. Radiation nephropathy is characterized by a progressive reduction in renal hemodynamics associated with a severe anemia. The latter is often normochromic normocytic in character, but can progress to a microangiopathic hemolytic anemia. The pathogenic mechanisms responsible for the development of radiation nephropathy remain ill-defined. Experimental studies which allow serial determinations of functional, morphologic, and cell kinetic radiation-induced changes indicate that primarily glomerular but also tubular alterations occur in the primary stages of radiation nephropathy. Glomerular capillary endothelial cell loss is seen within several weeks of irradiation. Remaining endothelial cells exhibit increased permeability leading to a subendothelial transudate. Mesangiolysis also is observed. In contrast, podocytes appear to be relatively unaffected at this stage. The endothelial changes appear to resolve, but the mesangial lesions progress, with hypercellularity and/or hypertrophy, increased mesangial matrix, mesangial sclerosis, and ultimately, glomerulosclerosis. These mesangial changes are similar to those observed in other chronic glomerulopathies. Dietary protein restriction, corticosteroids, and ACE-inhibitors all can reduce the severity of experimental radiation nephropathy. PMID- 8714749 TI - The influence of 5-fluorouracil on the endothelium in small arteries. An electron microscopic study in rabbits. AB - 5-Fluorouracil (5-FU) is a widely used antineoplastic agent. 5-FU induced cardiotoxicity is a still relatively unknown side-effect of this drug. This phenomenon could be due to a direct cytotoxic effect on the endothelial cells. We tested this hypothesis in an experimental study in rabbits, by scanning or transmission electron microscopic evaluation of endothelium in small arteries (the central artery of the ear) after in vivo treatment with 5-FU. Both local and systemic effects of 5-FU on endothelium were studied 15, 30, 60 and 120 minutes after intra-arterial or intraperitoneal treatment. Perfusion fixation at physiological pressure and temperature was used in order to minimize damage to the endothelium during the preparation procedure. Eighteen rabbits weighing 2.5 3.0 kg were used, and 6 animals served as controls. The following parameters were evaluated: vessel wall and endothelial cell contraction, cell edema, cytolysis, occurrence of denuded areas, platelet adhesion/aggregation and fibrin formation. For the description of each parameter a scale of negative points was used. Irreversible cell damage was observed in 5-FU treated animals: disruption of the endothelial sheet and patchy exposure of the subendothelium, sometimes as a focus for thrombus formation. Our findings support the hypothesis that the thrombogenic effect of 5-FU secondary to its direct cytotoxic effect on endothelium might be one of the pathophysiological mechanisms behind 5-FU induced cardiotoxicity. PMID- 8714750 TI - In vitro formation of "urinary stones": generation of spherulites of calcium phosphate in gel and overgrowth with calcium oxalate using a new flow model of crystallization. AB - Calcium phosphate (CaP) has been detected in the majority of urinary stones containing predominantly calcium oxalate (CaOx). Therefore, crystal phases of CaP might play an important role with respect to the formation of urinary calcium stones in general. Very often, CaP found in stones or tissue of human kidney occurs in the shape of small spherulites. In this paper, we report on a new flow model of crystallization (FMCG), which has been used to generate spherulites of CaP in a gel matrix of 1% agar-agar at 37 degrees C from a supersaturated, metastable solution continuously flowing over the gel surface. Scanning electron microscopy (SEM), X-ray diffraction and microscopic Fourier transformed infrared spectroscopy (FTIR) revealed that the particles formed (diameter: up to 200 microns) consisted of a poorly crystalline core of carbonatoapatite which was partly surrounded by a well-crystallized shell of octacalcium phosphate (OCP) showing radially oriented sheet-like structures. Subsequently, CaOx was grown on these spherulites from a flow of a correspondingly supersaturated solution conducted over the gel matrix. It could be shown by SEM that growth of calcium oxalate monohydrate (COM) was characteristically induced by the OCP shell. Radial sheet-like forms of OCP were directly continued by COM showing a certain radial orientation. The model of crystallization in gel matrices applied here should be well-suited to simulate the process of urinary stone formation under in vitro conditions. PMID- 8714751 TI - Alterations in MDCK and LLC-PK1 cells exposed to oxalate and calcium oxalate monohydrate crystals. AB - Structural analysis of human kidney stones reveals the presence of cellular membranes and other cell fragments. Experimentally, calcium oxalate crystallization is facilitated when an exogenous nephrotoxin is given with ethylene glycol, thus providing cellular degradation products to act as heterogeneous nuclei. In this report, we tested whether oxalate alone could act as a cell toxin capable of producing damaged cells without the presence of an exogenous agent. Cultured LLC-PK1 and MDCK cells, when exposed to 1.0 mmol KOx, a concentration at the limit of metastability for calcium oxalate nucleation, were severely damaged as measured by specific lactate dehydrogenase (LDH) release in the spent media and by trypan blue exclusion. This effect was magnified by the addition of pre-formed calcium oxalate monohydrate crystals; the injury was significantly amplified when compared to exposure to oxalate alone. Scanning electron microscopy studies illustrated attachment of crystals to cells with loss of cell-to-cell and cell-to-substrate contact, as cells were released from the monolayer. In both oxalate and combined crystal-oxalate studies, more cells were released from the monolayer and exhibited considerably more damage when compared to controls. Oxalate, at the limit of metastability for calcium oxalate, is a cell toxin and can produce cellular degradation products. This effect is increased significantly by the addition of calcium oxalate monohydrate crystals. PMID- 8714753 TI - Forty years of neuropsychopharmacology. PMID- 8714752 TI - Heterogeneous nucleation of calcium oxalate crystals in mammalian urine. AB - It is generally recognized that calcium oxalate crystal formation in urine is induced by heterogeneous nucleation. However, there is no consensus as to the nature of the nucleation substrate. Evidence is provided in this paper that membranous cellular degradation products are the most likely candidates because they: (1) are ubiquitous in urine and urinary stones; (2) are found in close association with crystal deposits in the kidneys; and (3) can induce nucleation of crystals from a metastable solution of calcium oxalate in vitro and metastable urine in vivo. PMID- 8714754 TI - Dopamine receptors--the present state of research and perspectives. AB - The aim of the review is a brief description of the present status of research and perspectives in the field of multiple dopamine receptors, as well as of the importance of new discoveries for the treatment of schizophrenic psychoses and Parkinson's disease. Since the discovery of D1 and D2 dopamine receptors in the seventhies, evidence has accumulated for the existence of some new dopamine receptors. Among them the D3, D4 and D5 receptors, as well as their isoforms have been cloned. Molecular biology and pharmacology of these receptors have been exhaustively studied in recent years. These studies have prompted a number of hypotheses which open new perspectives for investigation into pathophysiology and therapy of schizophrenia and Parkinson's disease. PMID- 8714755 TI - 5-Hydroxytryptamine in the central nervous system. AB - The review summarizes the results of studies into the role of 5-HT as a neurotransmitter in the central nervous system. The biosynthesis, release, metabolism and distribution of 5-HT; classification and localization of 5-HT receptors; electrophysiological effects of 5-HT and its influence on second messengers; autoregulation of 5-HT release; effects of 5-HT on the release of other neurotransmitters and pituary hormones; behavioral effects induced by 5-HT receptor agonists and the functional interaction between different receptor subtypes have been described. Additionally, some data on functional changes in the 5-HT system in psychiatric disorders have been cited, and a possibility of clinical utilization of 5-HT-active related drugs in neuropsychiatric and other diseases has been reviewed. PMID- 8714756 TI - Recent advances in the GABA-A-benzodiazepine receptor pharmacology. AB - Gamma-aminobutyric acid (GABA) acts on pharmacologically and functionally distinct receptors. These sites designated GABA-A and GABA-B receptors, differ with regard to their ionic characteristic and pharmacological properties. The most important distinction is, that the GABA-A receptor is associated with chloride channel and with membrane recognition sites for benzodiazepines. During the past decade numerous studies have made it possible to obtain more detailed knowledge of the structure and properties of the GABA-benzodiazepine receptor complex, which is made up of at least two distinct sites designated as BZD-1 and BZD-2 receptors. Third type, designated as peripheral type is located in the mitochondrial membrane and may regulate a steroidogenesis in the CNS. Molecular cloning studies showed a heterogeneity of GABA-A receptors, which are composed of multiple subunits (alpha, beta, gamma, delta and ro), which form distinct isoreceptors. New classes of GABA-BZD agonists such as zolpidem act selectively upon certain isoreceptors thus showing characteristic pharmacological properties. Recent studies provided detailed information on the interaction of ethyl alcohol (Et-OH) with GABA-BZD receptor complex. The most important finding is, that there are Et-OH sensitive and Et-OH resistant GABA-A receptor isoforms. Recent evidence reinforces the possibility, that reduced activity of the brain GABA-ergic system is associated with mechanism of depression. PMID- 8714757 TI - Excitatory amino acid antagonists and the anticonvulsive activity of conventional antiepileptic drugs. AB - There is a great body of evidence, that excitatory amino acid antagonists, apart from their anticonvulsive properties per se, potentiate the protective activity of conventional antiepileptics against maximal electroshock-induced seizures in mice. It is worth stressing, that combinations of valproate with either CGP 37849 (a competitive NMDA antagonist) or dizocilpine (MK-801, a non-competitive NMDA antagonist), providing a 50% protection against maximal electroshock, resulted in no adverse effects, as measured in the chimney test (motor coordination) or passive avoidance task (long-term memory). On the other hand, valproate administered alone at its ED50, to protect against maximal electroshock, produced profound adverse effects. However, some NMDA antagonists (D-CPP-ene, memantine, procyclidine or trihexyphenidyl) did enhance the protection offered by common antiepileptics but these combined treatments were associated with considerable side-effects on motor coordination and long-term memory. Interestingly, ifenprodil (an antagonist of the polyamine site within the NMDA receptor complex) possessed some anticonvulsive activity against electroconvulsions but failed to enhance the antielectroshock efficacy of conventional antiepileptics. AMPA/KA receptor antagonists (NBQX and GYKI 52466), similarly to NMDA antagonists, potentiated the protective action of antiepileptic drugs against maximal electroshock and these combinations were generally devoid of unwanted effects. PMID- 8714758 TI - Receptor and voltage-operated ion channels in the central nervous system. AB - Ion entry into neurons occurs either through receptor-operated channels (ROC) or voltage-operated channels (VOC). The function of ROC depends crucially on the action of agonists, antagonists or compounds modulating particular types of receptors (GABA A, NMDA, Ach N receptors). The function of VOC is closely connected with the activity of protein kinases and the processes of phosphorylation of membrane proteins (K+, Na+, Ca2+ channels). Gamma aminobutyric acid (GABA) is the main inhibitory neurotransmitter in the vertebrate brain. The GABA A receptor is a oligomeric complex of multiple binding sites and chloride channel. This complex contains recognition sites for GABA, anxiolytics such as benzodiazepine, anxiogenic--beta-carboline, and convulsant such as picrotoxin. Chloride ion channel plays a crucial role in anxiogenic, anxiolytic and convulsant activities. Glutamic acid is the main endogenous neurotransmitter for N-methyl-D-aspartate (NMDA)-type excitatory amino acid receptor. NMDA receptors connected with Ca2+ channel, have multiple modulatory sites which are affected by a wide range of compounds. There are NMDA and competitive NMDA antagonists site, the glycine site, the phencyclidine (PCP) site and the binding site of Mg2+ ions in this receptor complex. Calcium entry through NMDA receptors may be important in the etiology of many psychiatric disorders. VOC mediate rapid, voltage-gated changes in ion permeability during action potentials in neurons. Electrophysiological studies indicate the existence of three types of VOC (K+, Na+, Ca2+ channels). In number of neurons various subtypes of Ca2+ channels (P, T, N and L-type) occur together. Among them, the L-type calcium channel has been first described and most thoroughly studied. The L-type calcium channel is localized on nerve terminals in the pre and postsynaptic parts, as well as on cell bodies and may be involved in the mechanism of action of psychotropic drugs. Chronic treatment with various psychotropic drugs changes the density of voltage dependent Ca2+ channels in the central nervous system. Thus calcium entry through both VOC and ROC may be important in the etiology of many psychiatric disorders. PMID- 8714759 TI - Metal sensitivity of some symbiotic N2-fixing bacteria and Pseudomonas strains. AB - An investigation was carried out to determine the sensitivity of different soil microbes (Rhizobium, Bradyrhizobium and Pseudomonas) to various metals (Cu2+, Zn2+, Co2+, Mn2+, Mo2+ and Fe2+) in vitro. Sulphate and chloride forms of these microelements were used (except Mo2+ as Na2Mo04) in 0.1, 1.0 and 10 micrograms/ml concentrations in modified YEM and nutrient broth. Growth (optical density, OD550 and OD640) of bacterium inoculated (approx. 10(6) CFU/ml) tubes, was measured spectrophotometrically after 48 h of incubation of 28 degrees C in a rotary shaker (150 rpm). Data of triplicate samples are shown as percent of control tubes (inoculated, free from treatments) and after an analysis of variance SE was calculated. Strains of Rhizobium leguminosarum proved to be the most sensitive to Cu2+, Zn2+ and Co2+. The slow growing Bradyrhizobium and plant growth promoting (PGPR) Pseudomonas isolates, however, were affected only at the highest (10 micrograms/ml) dose of these elements. In contrast Mn2+, Mo2+ and Fe2+ microelements were stimulatory for the growth of all investigated soil microbes. Sulphate forms of the most harmful Cu2+ and Zn2+ cations were more toxic than the chloride forms. An especially high diversity was found among the R. leguminosarum bv. viceae isolates. Monitoring the sensitivity of these microbes has a primary importance for selection of ecologically diverse isolates, as potential inocula in heavy-metal affected soils. PMID- 8714760 TI - Some environmental factors influencing the survival of Rhizobium leguminosarum bv. viceae. AB - Investigations were carried out to monitor the sensitivity of Rhizobium leguminosarum bv. viceae strains to various environmental stress-factors (salinity, pH, Al3+, Ca2+, Mg2+) and select them as potential candidates for Vicia faba inoculation. In the range between pH 5.0 and 10.0, the salt effect of 10-500 mM NaCl, Ca2+, Mg2+ (as chlorides and sulphates), Al2O3 and KAl (SO4)2 (25 400 microM) were tested in modified yeast-mannitol (YEM) liquid medium. Cell density of the suspensions inoculated with R. leguminosarum bv. viceae strains (initial cell-number 10(6) CFU/ml) were measured spectrophotometrically after 48 h incubation in a rotary shaker (rpm 150) at 28 degrees C. Data of optical density (OD550) are shown as percent of control (inoculated, free from treatments, pH 7.0) tubes. It was established, that results of tolerancy were in agreement with those found earlier for Rhizobium sensitivity. Strain of Lobab Z (isolated in Hungary) however proved to have an especially outstanding survival in any media tested in vitro. Lowest observed effect concentrations (LOECs) were: 500 mM NaCl, 200 microM Al3+ (as Al2O3 or KAl (SO4)2), 50-100 mM Mg2+, and 200 300 mM Ca2+. For the Al3+, Ca2+ and Mg2+ cations, there was no variation between the stress-effect of sulphate and chloride anions. Both forms of Ca2+, however significantly reduced the growth potential of R. leguminosarum bv. viceae strains. PMID- 8714761 TI - The effect of dimethyl sulphoxide on CCl4-induced damage to the liver and its effects on hepatic glutathione and glucose. AB - This investigation was undertaken to determine if an interaction of toxicologic importance might occur during a prolonged exposure of rats to carbon tetrachloride (CCl4) and Dimethyl Sulphoxide (DMSO). CCl4 administration produced a significant decrease in hepatic microsomal glucose-6-phosphatase activity accompanied by a small increase in alkaline phosphatase activity, Glutathione depletion was highest when CCl4 was administered alone. DMSO, did not increase hepatic uptake of glucose. These findings suggest that DMSO given at low dose can prevent the decrease of hepatic glucose-6-phosphatase but may indirectly affect the level of tissue glucose. PMID- 8714762 TI - Effects of an organophosphate on the antioxidant systems of fish tissues. AB - The effect of a Hungarian made and applied insecticide, organophosphate (Dichlorvos) on antioxidant enzymes and other oxidative and redox parameters of two different fish species, carp (Cyprinus carpio L.) and catfish (Ictalurus nebulosus) was studied. The two fish species have different ways of life. From antioxidant enzymes, changes in superoxide dismutase, catalase, glutathione peroxidase and, in the case of carp, acetylcholinesterase activities were studied in tissue homogenates. Other parameters studied: changes of lipid peroxidation, reduced glutathion and two radicals, which are superoxide and hydroxyl radicals. Our results showed that the organophosphate tested, besides its inhibitory effect on acetylcholinesterase-or together with it-induces changes characteristic of "oxidative stress". PMID- 8714763 TI - High efficiency callus induction and plant regeneration in petiole culture of four poplar genotypes. AB - In order to develop a simple and feasible approach to achieve high frequency plant regeneration for protoplast isolation and transformation experiments, a method was elaborated by using a new type of explant (petiole segments) for the four Populus nigra genotypes. Callus initiation from the petioles took place on N6 medium containing 2,4-D (0.1-1 mg/l). The highest rate of callus initiation (100%) was achieved when the basic medium was supplemented with 0.5 mg/l of 2,4 D, in all tested genotypes. For shoot regeneration, calli were transferred to MS and WPM medium supplemented with BA (1.0-2.5 mg/l) and NAA (0.2 mg/l). Multiple shoot regeneration was observed in each shoot induction medium. The highest rate of shoot regeneration (6.83 shoot/callus) was observed on MS medium containing 2.5 mg/l BA and 0.2 mg/l NAA. The results showed highly significant differences between the media. There was no significant difference between the genotypes and genotype x medium interaction. PMID- 8714765 TI - Effects of 6-hydroxydopamine on testosterone production by mouse leydig cells in vitro. AB - Increasing doses (15 to 1000 micrograms/ml) of 6-hydroxydopamine (6-OHDA) stimulated, then suppressed testosterone production by mouse leydig cells incubated for 3 h in vitro. The stimulating doses ranged between 50 and 100 micrograms/ml, with maximal effects occurring at 30 to 60 minutes after the start of the treatments. At doses of 500 micrograms/ml, 6-OHDA exhibited inhibitory effects. When added to leydig cell incubations together with stimulating doses of luteinising hormone (LH), 1-(3',4'-dihydroxyphenil)2-isopropylaminoethanol (L isoproterenol) or 8-bromoadenosine 3':5'-cyclic monophosphate (8-Br-cAMP), 6-OHDA abolished the effects of the latter compounds. Prolactin and prostaglandin E2 (PGE2) inhibited the stimulating effects of 8-Br-cAMP but not LH. It is proposed that the actions of 8-OHDA affect intracellular sites yet to be identified, thereby inhibiting testosterone production by mouse Leydig cells. Some of the actions of 6-OHDA seem to be medicated via beta-adrenergic receptors as the latter abolishes the stimulatory effects of L-isoproterenol, a potent beta adrenergic agonist. However, the inability of stimulatory doses of LH and 8-Br cAMP to reverse the inhibitory effects of 6-OHDA point to the possibility that other actions of 6-OHDA may be relayed via a second messenger system separate from that involving cAMP. PMID- 8714764 TI - Role of cholinergic receptors in melanophore responses of amphibians. AB - Responses of isolated skin melanophores of Rana tigerina and Bufo melanostictus to cholinergic drugs were studied using the Mean Melanophore Size Index assay to explore the nature and role of cellular receptors in melanophore regulation activity. Acetylcholine (Ach) in a concentration of 10(-7) to 6.4 x 10(-6) g/ml caused dispersion of the skin melanophores of R. tigerina and B. melanostictus. These effects were blocked by both atropine and hyoscine in a concentration of 8 x 10(-6) g/ml each. Eserine augmented the melanophore dispersal effects of Ach. This potentiation of the dispersal effect of Ach by eserine was also antagonised by hyoscine. Carbachol another specific cholinergic agonist significantly caused dispersion of the melanophores of both the amphibian species. The effects were also blocked by atropine and hyoscine. These data indicate that cholinergic receptors of muscarinic type are present on the melanophores of R. tigerina and B. melanostictus which mediate dispersion of integumental melanophores leading to darkening of the skin. PMID- 8714766 TI - Ultrastructural variations of the rat fetal adrenal cortex after maternal bilateral adrenalectomy and exogen glucocorticoid administration. AB - Rat fetal adrenal cortex ultrastructure was studied in fetuses of 20 days of gestation coming from control (C) pregnant rats and from mothers undergoing two experimental conditions: maternal bilateral adrenalectomy on day 1 of gestation (ADX) and maternal bilateral adrenalectomy plus posterior dexamethasone administration the days 15, 16 and 17 of gestation (A+D). Our results show ultrastructural changes in the ADX rat fetal adrenal cortex, consisting of hyperactive steroidogenical features as response to the absence of maternal glucocorticoids, and the restoration of the control structural characteristics after dexamethasone administration to adrenalectomized animals. PMID- 8714767 TI - Similar localization of immunoreactive glutamate and aspartate in the pineal organ and retina of various nonmammalian vertebrates. AB - The localization of immunoreactive glutamate and aspartate was compared in the pineal organ and retina of various vertebrates (Raja clavata, Carassius auratus, Salvelinus alpinus, Triturus vulgaris, Triturus cristatus, Lacerta muralis, Lacerta agilis, Lacerta viridis, Columbia livia and white leghorn chicken) by postembedding immunoelectron microscopy. Immunoreaction of both excitatory amino acids was detected in the pinealocytes in a localization similar to that of retinal photoreceptors. The reaction was intense in the axonal processes of pinealocytes as well as retinal rods and cones, further in their terminals on secondary pineal and retinal neurons. Subsequent immunoreaction on the same section showed a colocalization of glutamate and aspartate. The accumulation of these amino acids in the presynaptic part of pinealocytes suggests that they act as synaptic mediators in the neural efferentation of the pineal organ. In reptiles and birds where the hormonal efferentation of the pineal is well developed, glutamate and aspartate was also found to be accumulated in neuroendocrine terminals of pinealocytes. Therefore, glutamate and aspartate may have a role in both the hormonal and neural efferentation of the pineal organ. PMID- 8714768 TI - Testicular morphology and testosterone levels in dogs immunized with antibodies generated against an androgen binding antigen in the epididymal cauda. AB - Adult dogs (Alsatian crosses) were passively immunised intrascrotally or intramuscularly with antibodies generated against an epididymal cauda androgen binding antigen (CABA) for 10 days. The testes were examined histologically 10 or 30 days posttreatment. Testosterone levels were determined on blood obtained daily for 10 days before, during and after treatment. A dose-dependent exfoliation of sperm, spermatids and spermatocytes, in that order, was observed in immunised animals. This was concomitant with the ballooning of luminal ends of Sertoli cells. The effects were, however, reversible. The Leydig cells were not affected by the treatments and plasma testosterone levels before, during and after treatment remained unchanged. It is concluded that an antigen similar to CABA is present in the testis, the neutralisation of which causes testicular lesions without interfering with testosterone levels. PMID- 8714769 TI - The structure of the dart and dart sac of the snail Eremina ehrenbergi, Roth. 1839. AB - The dart of Eremina ehrenbergi is a small white delicate structure. It consists of funnel-shaped corona with two blades; a short flare and an extended shaft which is surrounded by two lateral vanes. Dart formation was investigated by examining the histological structure of the dart sac of snails before and after copulation. Dart sac is a finger like muscular protrusion. Its internal epithelial lining forms at its blind edge an extended tubercle which gives its lumen a Y-shape. Dart formation begins at the tip of the tubercle in which the epithelial cells secrete a crystallite and homogeneous material which forms the mineralized part of the corona. The subsequent secretory activity of the tubercle epithelium results in an increase in the diameter and anterior lengthening of the corona and the flare. The continued secretory activity of the dart sac epithelium forms the layers of the shaft and the lateral vanes. The fine sculpturing structure of the dart had been investigated by using SEM. PMID- 8714770 TI - Proteins of bovine leukemia virus and human T-cell leukemia viruses. PMID- 8714771 TI - Natural human interferon-alpha enhances the expression of intracellular adhesion molecule-1, integrin alpha 2 and beta 1 by a mucosal epithelial cell line. AB - We have found that pretreatment of a human oral mucosal epithelial cell line KB with natural human interferon-alpha (IFN-alpha) augments the expression of intercellular adhesion molecule-1 (ICAM-1), and the complex of CD29 and CD49b, which is known as very late antigen (VLA)-2, in a dose-dependent manner, whereas the expression of other adhesion molecules such as CD44 leukocyte function associated antigen-3 (LFA-3), VLA-4 and endothelial leukocyte adhesion molecule-1 (ELAM-1) did not show any significant changes under the same conditions. The pretreatment of KB cells with IFN-alpha also induces the adhesion of these cells to the human leukemia T cell line MOLT-16 and to peripheral T lymphocytes in a dose-dependent manner. However, the adhesion of the above-mentioned T cells to KB cells was not inhibited by specific antibodies against ICAM-1, CD29, and CD49b. The results thus show that adhesion molecules other than ICAM-1, CD29, AND CD49b are responsible for the induced adhesion between T cells and IFN-alpha-pretreated KB cells. PMID- 8714772 TI - Distribution of gamma delta T cells in the pig foetus. AB - The distribution of gamma delta T cells during different developmental stages of pig foetuses was studied using monoclonal antibodies directed against a ruminant WC1 antigen, a T-cell receptor (TCR) subset epitope, and pig Null T-cell antigen. Binding was revealed by the avidin/biotin/peroxidase technique on cryostat sections and flow cytometry analysis. The extrathymic origin of gamma delta T cells was confirmed: gamma delta T cells appeared in the liver, spleen and peripheral blood sooner than in the thymus. In the course of the second half of gestation, these T cells were found in the dermis and intestinal mucosal compartments which represented the predominant loci of gamma delta T cells after birth. PMID- 8714773 TI - Dual role of fibroblasts in tumor cell growth. AB - Vast experience with cultivating biopsies from human tumors indicates that in most cases the admixture of fibroblasts has a negative effect on growth of tumor cells. Only rarely is observed help provided by the fibroblasts. It has also long been known that fibroblasts can inhibit by contact themselves and also produce growth factor(s) promoting cell proliferation. We have used three permanent squamous cell carcinoma lines and fibroblasts derived from biopsies of trachea to study this paradox. The inhibitory activity of fibroblasts is contact-dependent in a cell membrane-bound factor, which is trypsin sensitive. We prepared microsomal fractions (MF) from aged (more than 40 passages) fibroblasts and followed their effect on proliferation of the tumor cell lines in MTT assay. MF from all three fibroblast lines inhibited the tumor cells. Most regularly was this phenomenon observed with line UM-SCC 22B. Not all tumor cells are immortal. On the contrary, a large portion of them undergoes terminal differentiation. With the line UM-SCC 22B we tested the possibility that MF from fibroblasts can increase the portion of tumor cells which will senescence after few mitoses. The immortal cells were defined as cells capable in 8 or 13 days of forming colonies of more than 50 or 200 cells. The senescent cells were defined as cells not capable of producing within the same period of time colonies of more than 12 or 30 cells. The MF was able to increase the number of small colonies, i.e. the number of senescent tumor cells. The fibroblasts of the same passage level were releasing soluble growth factor(s) promoting growth of cells. Fibroblasts can apparently simultaneously inhibit growth by contact and release factor(s) promoting growth of cells. The final outcome is a result of the balance between these two forces. This balance is regulated by many intrinsic and extrinsic forces. PMID- 8714774 TI - Haemopoiesis-enhancing effects of repeatedly administered carboxymethylglucan in mice exposed to fractionated irradiation. AB - Carboxymethylglucan (CMG), a water-soluble glucan derivative, enhanced the number of granulocytes in the peripheral blood as well as other indices of haemopoietic recovery (total cellularity and the number of granulocyte-macrophage progenitor cells in femoral marrow, spleen weight) investigated after fractionated gamma irradiation in mice (five doses of 2 Gy each, or three, four and five doses of 3 Gy each given at 24-h intervals). An increased liver weight and a more pronounced anaemia found in the CMG-treated mice suggested, however, that also inflammatory side effects were evoked by the repeated administration of CMG. On the other hand, the development of tolerance, i.e., a decreased effectiveness of the treatment with CMG upon its repeated administration, did not seem to play any major role under the experimental conditions studied, because the protective effects of CMG increased with the increasing number of CMG injections. PMID- 8714775 TI - Effect of antituberculous drugs, isoniazid, pyrazinamide and rifampicin, on chemiluminescence of the human polymorphonuclear leukocytes. AB - Luminol-dependent chemiluminescence (CL) was used to assay the effect of three antituberculous drugs [isoniazid (INH), pyrazinamide (PZA), rifampicin (RM)] on the production of reactive oxygen species (ROS) by polymorphonuclear leukocytes (PMN) isolated from blood of 5 healthy donors. Drugs were added directly to the medium in concentrations; INH 5 micrograms/ml, PZA 40 micrograms/ml, RM 7 micrograms/ml. These concentrations correspond to peak serum concentrations after usual doses. RM was found to inhibit FMLP (N-formyl-methionyl-leucyl phenylalanine)-induced CL response of PMN, PZA did not influence the metabolic activity of neutrophils, and INH stimulated PMN CL. PMID- 8714776 TI - Development of the human striatum: implications for fetal striatal transplantation in the treatment of Huntington's disease. AB - Fetal neural transplantation has recently been demonstrated to ameliorate motor and other behavioral deficits in animals models of Huntington's disease, and reconstruct many of the damaged striatal circuits. However, there has been significant variability in the histological appearance of these grafts, most likely related to differences of the regions of dissection of the donor tissue. Selective dissection and transplantation of the lateral ventricular eminence in rodents has resulted in grafts consisting of primarily striatal-like tissue. This data, combined with data from our own and other laboratories has led to a description of the development of human striatum, with a particular emphasis on the relevance of human striatal development to the field of fetal tissue transplantation for the treatment of Huntington's disease. If the goal of transplantation is to graft GABAergic striatal projection neurons, it is our impression that optimal grafting results will occur when transplants are derived from the lateral ventricular eminence and the lateral aspect of the body of the ventricular eminence anterior to the foramen of Monro. Optimal results are likely to occur when donor ages range from Stage 19 to 23, with possible graft success when donor age extends to as late as postovulatory week 22. PMID- 8714777 TI - Sources of human hematopoietic stem cells for transplantation--a review. AB - Transplantation of hematopoietic stem cells provides a means of replacing a defective hematopoietic system in patients with a wide range of malignant and nonmalignant disorders that affect the blood forming tissue. The same procedure has also allowed dose-escalation of standard chemotherapy and radiotherapy in the treatment of malignant disease of nonhematological origin. Unit recently, bone marrow has been the sole source of hematopoietic stem cells, but limitations of conventional bone marrow transplantation have stimulated a search for alternative sources and uses of stem cells. Fetal tissues (especially liver) are a recognized source of transplantable stem cells and offer the great advantage of reduced immunogenicity, potentially removing the problems of tissue type matching. Umbilical cord blood is also a rich source of stem cells and, although it contains alloreactive cells, it is readily available without special ethical constraints. Both fetal tissue and cord blood suffer the disadvantages of limited numbers of stem cells per donation, and there is much interest in the development of technologies for the safe and reliable expansion and/or pooling of stem and progenitor cells. The observation that small numbers of stem cells are found in the peripheral blood of adults has led to the exploitation of the blood as a further source of stem cells. The ability to mobilize these cells from the medullary compartment into the periphery by the use of chemotherapy and/or recombinant hematopoietic growth factors has enabled the collection of sufficient numbers of cells for transplantation purposes. All of these advances are increasing the options and the range of choices available to clinicians and patients in the arena of hematopoietic stem cell transplantation. PMID- 8714778 TI - Effects of hibernation or cryopreservation on the survival and integration of striatal grafts placed in the ibotenate-lesioned rat caudate-putamen. AB - Tissue storage prior to intracerebral transplantation would represent a major advantage when conducting clinical transplantation trials in that the procurement of the embryonic donor tissue and the timing of neurosurgery could be planned more efficiently. In the present study, the effects of storing rat embryonic striatal tissue at either +4 degrees C or below freezing temperature prior to grafting to the adult striatum, were assessed with regard to transplant survival, morphology and integration. Eleven days following a unilateral injection of ibotenic acid into the head of the caudate-putamen, a control group of rats received grafts of striatal primordium prepared immediately after dissection from rat embryos (embryonic day 16). A second group of rat embryonic striatal tissue was stored at 4 degrees C (hibernation) for 5 days and then transplanted. A third group of the striatal donor tissue was cryopreserved in liquid nitrogen for 5 days before implantation surgery. Six to seven weeks following transplantation surgery, the grafts were analysed in brain sections processed for acetylcholinesterase histochemistry, DARPP-32 (dopamine and cyclic AMP regulated phosphoprotein with a molecular weight of 32 kDa) and tyrosine hydroxylase (TH) immunocytochemistry. The mean total graft volume and the relative size of the AChE-positive regions were not significantly different between the three groups. Striatal-specific graft regions, positively stained for AChE and DARPP-32, generally exhibited TH immunoreactivity, suggesting that they had received dopaminergic afferents from the host brain. We conclude that embryonic rat striatal tissue can be cryopreserved or hibernated over 5 days without significant impairment in the yield of striatal neurons following intrastriatal implantation and without markedly affecting transplant morphology. PMID- 8714780 TI - Phenotypic reversion of smooth muscle cells in hybrid vascular prostheses. AB - Our purpose was to evaluate whether or not and when phenotypic modulation of smooth muscle cells (SMCs) in hybrid vascular prostheses preincorporated with SMCs occurs upon implantation. Two types of hybrid vascular grafts incorporated with vascular cells derived from canine jugular veins were prepared: grafts containing a collagen gel layer covered with an endothelial monolayer at the luminal surface (Model I graft) and those containing an endothelial monolayer and SMC multilayer (Model II graft). They were bilaterally implanted into carotid arteries of the same dogs from which the cells had been harvested for 2 wk (n = 3) and 12 wk (n = 3). The time-dependent changes in populations of three SMC phenotypes (synthetic, intermediate, and contractile) in the neoarterial layers were quantified by morphometric evaluation using a transmission electron microscope in hybrid vascular grafts. Before implantation, all the SMCs were of the synthetic phenotype. In Model II grafts at 2 wk, synthetic and intermediate SMCs were dominant especially in the luminal layer. On the other hand, neoarterial layers at 12 wk were dominated by contractile SMCs, which were evenly distributed throughout the entire neoarterial tissues. A markedly delayed phenotypic reversion was noted for the Model I grafts at 12 wk. In the hybrid grafts, during about 3 mo of implantation, neoarterial SMCs transformed from the synthetic to the contractile phenotypes, which was promoted by SMC incorporation. PMID- 8714779 TI - Effective cryopreservation and long-term storage of primary human hepatocytes with recovery of viability, differentiation, and replicative potential. AB - Despite reports of successful cryopreservation of primary human hepatocytes, existing methods do not produce sufficient recovery of viable cells to meet the needs of basic research or clinical trials of hepatocellular transplantation. We now describe a protocol for efficient cryopreservation of primary human hepatocytes using University of Wisconsin (UW) solution, fetal bovine serum, and dimethyl sulfoxide (DMSO). This method provides > 90% viability of differentiated, primary human hepatocytes 8 mo after cryopreservation as measured by trypan blue exclusion, preserves hepatocyte morphology, liver-specific gene expression (alpha 1 antitrypsin), and replication. The effectiveness of UW solution as a cryopreservative agent suggests that metabolic as well as ultrastructural factors may be important in the effective cryopreservation of primary human hepatocytes. The present method represents an effective protocol for cryopreserving differentiated primary human hepatocytes for research. This method may allow characterization and banking of human hepatocytes for clinical applications, including hepatocellular transplantation and hepatic assist devices. PMID- 8714781 TI - Self-organized, tubular hybrid vascular tissue composed of vascular cells and collagen for low-pressure-loaded venous system. AB - A tubular, hierarchically structured hybrid vascular tissue composed of vascular cells and collagen was prepared. First, a cold mixed solution of bovine aortic smooth muscle cells (SMCs) and Type I collagen was poured into a tubular glass mold composed of a mandrel and a sheath (example of dimensions: inner diameter, 1.5 mm; outer diameter, 7 mm; length, 7 cm). Upon incubation at 37 degrees C, an SMC-incorporated collagenous gel was formed. After the sheath was removed, the resulting fragile tissue, when cultured in medium, thinned in a time-dependent manner to form an opaque, dense tissue. Higher SMC seeding density and lower initial collagen concentration induced more rapid and prominent shrinkage of the tissue. Morphologic investigation showed that over time, bipolarly elongated SMCs and collagen fiber bundles became positioned around the mandrel. Both components became circumferentially oriented. When the mandrel was removed, a tubular hybrid medial tissue was formed. A hybrid vascular tissue with a hierarchical structure was constructed by seeding endothelial cells onto the inner surface of the hybrid medial tissue. Prepared tissues tolerated luminal pressures as great as 100 mmHg and mechanical stress applied during an anastomotic procedure. This method allowed us to prepare a tubular hybrid medial tissue of predetermined size (inner diameter, wall thickness, and length) by selecting appropriate mold design, initial collagen concentration, and SMC seeding density. Such hybrid vascular tissues may provide physiological functions when implanted into the venous system. PMID- 8714782 TI - Amount and distribution of collagen in pancreatic tissue of different species in the perspective of islet isolation procedures. AB - Because collagen is the major target in the enzymatic dissociation of the pancreas for islet isolation, we determined the amount of collagen and its distribution in a comparative study comprising normal pancreata of rat, dog, man, young pig, and adult pig. Collagen content was determined using a colorimetric method and its distribution was assessed in tissue sections stained with Sirius red. The collagen content is relatively low in the rat and adult pig pancreas, and the amount of collagen is relatively low in the septa of the rat and dog pancreas. Not the amount of collagen in the septa but collagen in the rest of the pancreas, mainly located between the acini, seems to determine the dissociation of the pancreatic tissue. This can be exemplified by the higher islet yields obtained from the adult vs. the young pig pancreas; the latter contains a higher total amount of collagen but a similar, relatively high, amount of collagen in the septa. A high amount of collagen surrounding the islets seems to be of secondary importance in islet isolations, because yields of the same magnitude are obtained from the canine and human pancreas containing a relatively low vs. high amount of collagen around the islets but a similar total collagen content. The rat pancreas contains both a low total amount of collagen and a high amount of collagen around the islets; therefore, the general experience that islet isolation procedures are effective in rats can be readily understood. PMID- 8714783 TI - The effect of UW solution and its components on the collagenase digestion of human and porcine pancreas. AB - University of Wisconsin (UW) solution is used extensively as a cold storage solution during the procurement and transport of the pancreas prior to islet isolation. However, it has been observed that UW inhibits the collagenase digestion phase of human but not porcine islet isolation, resulting in poor islet yields and islets of poor viability. The aim of this study was, therefore, to confirm this species difference and to determine which components of UW are responsible for the inhibition in the human. In the initial experiment, blocks of human and porcine pancreas (n = 7) were incubated in test tubes containing collagenase at a concentration of 4 mg/mL at 37 degrees C dissolved in 4 mL of either Hanks' solution or UW. Every 5 min the tubes were manually shaken and the degree of tissue dissociation scored on a scale of + and 3+. Our results confirm the inhibition of collagenase digestion in the human but not the pig. Using the same methodology, we then investigated the components of UW that were causing the observed inhibition in the human pancreas (n = 7). This time the collagenase was dissolved in individual or combinations of UW components. Using Hank's as a control, the results were then expressed as a median ratio. The components found to be most inhibitory were magnesium, the Na+/K+ ratio, hydroxyethyl starch (HES), and adenosine. Allopurinol in combination with either lactobionate or glutathione was markedly inhibitory (i.e., median ratio 1.8 and 1.9, respectively). The most inhibitory solution tested was combination of the three components raffinose, glutathione, and lactobionate (median ratio 2.1). This combination was almost as inhibitory as UW itself (median ratio 2.7). These findings are essential for the development of effective cold-storage solutions for the human pancreas that do not inhibit the subsequent collagenase digestion phase of islet isolation. PMID- 8714784 TI - A method for mass harvesting islets (Brockmann bodies) from teleost fish. AB - In certain species of fish, the insulin-producing tissue is uniquely located in separate structures called Brockmann bodies (BBs). Tilapia BBs have been shown to be a simple and inexpensive source of islet cells for xenotransplantation research. Each donor tilapia contains roughly 12-15 BBs, measuring from 0.3 to 5.0 mm in maximum dimension, in a triangular region of adipose tissue bounded by the liver, stomach, and spleen/gallbladder. At present, the larger BBs (usually 2 4) are harvested by microdissecting these "BB regions" using jeweler's forceps and microvascular scissors while being visualized with the aid of a dissecting microscope. It is a simple but time-consuming task that would not be applicable for harvesting massive amounts of BB tissue for large animal studies. Therefore, we have developed an easier and more efficient method of harvesting BBs based on a standard enzymatic method for isolating human adipocytes. BB regions are harvested from donor fish and pooled into a 50 mL plastic tube containing collagenase Type II (3 mg/mL) in Hank's balanced salt solution (HBSS); the tube is then placed in a 37 degrees C waterbath/shaker for roughly 15 min. The exact length of the digestion interval is determined by visual inspection of the tube to determine whether the BBs have been liberated. The digestion is then stopped by adding excess cold HBSS. The adipocytes float while the BBs and residual connective tissue (i.e., a few blood vessels, nerves, and bile ducts) form a pellet. The pellet is washed several times in HBSS and then placed in a culture dish. The BBs are easily handpicked with a siliconized pipette. Based on functional data and DNA content, this new method roughly doubles or triples our yield of BB tissue per donor fish. To determine whether BBs harvested in this manner functioned in a manner similar to those harvested by microdissection, we performed a series of transplants using mass-harvested BBs. Long-term normoglycemia was achieved in streptozotocin-diabetic nude mice and mean graft survival time was not altered in streptozotocin-diabetic euthymic balb/c mice. However, the total weight of donor fish required per recipient was decreased by 50% in both strains. PMID- 8714785 TI - Anti-plasticizer specific IgE is present in the serum of transfused patients. AB - Sera from 29 patients who had reacted to a platelet (27) or packed red cell (2) transfusion and from 5 patients who had received platelets without reacting were collected over a 13-month period. The sera were assayed for total IgE, and IgE specific for ethylene oxide, phthalic anhydride, hexamethylene diisocyanate, methylene diphenyl diisocyanate, toluene diisocyanate, and mast cell tryptase. Three patients with reactions had elevated total IgE levels, and specific IgE was positive for hexamethylene diisocyanate in 2 of 28 (7.1%), methylene diphenyl diisocyanate in 1 of 29 (3.4%), and toluene diisocyanate in 14 of 27 (51.9%). No positives were found in patients without reactions and no patients had an elevated tryptase level. It is unlikely that anti-plasticizer hypersensitivity was responsible for the transfusion reactions, but the prevalence and significance of such antibodies in both the hospital population and the general population would merit further investigation. PMID- 8714787 TI - Influence of applied brachial recording forces on pulse wave velocity and transmission in the brachio-radial arterial segment. AB - Arterial stiffness in hypertension and heart failure may increase afterload on the left ventricle. Pulse wave velocity and transmission ratio measurements are noninvasive methods to assess arterial stiffness. Since noninvasive pressure pulse recording requires sufficient applied force to distort the vessel wall, we hypothesized that the pulse wave velocity, transmission ratios, and distal pulse amplitudes and contours may be altered by the recording technique. Brachial and radial arterial pressure pulses were recorded simultaneously using a piezoelectric pulse transducer in 14 young, normal, male subjects using 10 brachial artery recording forces (0.35-3.58N, approximately equal to 0.36N increments) applied in a random order and a constant radial force (2.35N). Pulses were Fourier analyzed. One subject was excluded from analysis because of improper transducer positioning over the brachial artery. In 8 subjects, no significant changes occurred in any variable over all brachial recording forces. In the remaining 5 subjects, the measured variables remained constant until brachial artery recording forces exceeded 2.42 +/- 0.03N. The pulse wave velocity (p < 0.04), transmission ratios (harmonics 2-5, p < 0.0001), radial pulse amplitude (p < 0.0003), and relative powers (harmonics 2-5, p < 0.02) then decreased. In these subjects, brachial artery depths were less than the other subjects (5.9 +/- 0.4 vs. 7.7 +/- 0.4 mm, p < 0.05). The brachial and radial artery recording forces normally used during clinical measurements by 2 investigators were 1.43 +/- 0.01N (95% confidence intervals (CI) = 1.23N, 1.62N) and 1.88 +/- 0.11N (95% CI = 1.65N, 2.10N), respectively. Therefore, at forces normally used by clinical investigators, the pulse wave velocity, harmonic transmission ratios, and pulse amplitudes and contours obtained at the brachial and radial artery are not significantly influenced by forces applied at the brachial artery. However, these variables may be decreased in subjects with more superficial arteries when higher recording forces are used. PMID- 8714786 TI - Urinary angiotensin I-converting enzyme activity is increased in experimental acute renal failure. AB - The angiotensin I-converting enzyme (ACE) activity was studied in 2 experimental models of acute renal failure: (a) rats treated with a single injection of mercuric chloride (1.5 mg/kg) and (b) rats treated with a single injection of potassium dichromate (15 mg/kg). Rats were sacrificed 24 and 48 h after mercuric chloride or potassium dichromate injection. ACE activity was measured in urine, serum, and kidney. These data were compared with vehicle-treated rats. Rats with acute renal failure had proteinuria, polyuria, and decreased creatinine clearance. The damage to the kidney proximal tubule was evident by (a) the histological analysis at light and electron microscopy, (b) the augmentation in the urinary excretion of dipeptidyl aminopeptidase IV and N-acetyl-beta-D glucosaminidase, and (c) the low molecular weight proteinuria pattern. In addition, the histological analysis at the ultrastructural level showed normal glomeruli appearance. The above data suggest that the increased urinary excretion of enzymes and proteins in rats with acute renal failure is a consequence of tubular injury. Urinary and serum ACE activities increased and kidney ACE activity decreased. Our data suggest that the increase in urine ACE activity may be due to the kidney proximal tubule damage. This work supports the contention that an increase in urine ACE may be an indicator of injury to the proximal tubule. PMID- 8714788 TI - Relation of cholesterol esterification rate to the plasma distribution of high density lipoprotein subclasses in normal and hypertensive women. AB - We studied the particle size distribution of plasma high-density lipoproteins (HDL) by gradient gel electrophoresis and by assay of cholesterol esterification rate (FERHDL) in plasma depleted of very low (VLDL) and low-density (LDL) lipoproteins in 32 hypertensive women (53 +/- 10 y old) and in an age-matched group of 21 apparently healthy women. There were no significant differences between the groups with respect to their plasma total, HDL- and LDL-cholesterol. The plasma triglyceride (TG) concentration was significantly higher in the group of hypertensive women, and HDL-free cholesterol was significantly lower in the hypertensive group. There were highly significantly differences in the relative proportions of HDL subclasses between the hypertensive and control women: HDL2b was significantly lower and HDL3b,c was significantly higher in hypertensive patients compared to controls. FERHDL was 12 +/- 4%/h in the control group and 18 +/- 6%/h in patients with hypertension (p < 0.001). There was a strong positive correlation between FERHDL and the relative content of HDL3b,c, and a strong negative correlation between FERHDL and HDL2b in both groups. Comparison of subgroups of healthy and hypertension pre- and postmenopausal women revealed, apart from age, no significant differences between the healthy subjects in any of the above parameters. PMID- 8714789 TI - Cost analysis of dialysis treatments for end-stage renal disease (ESRD). AB - The cost of alternative dialysis modalities for the treatment of end-stage renal disease (ESRD) was evaluated, using a societal viewpoint, in a regional nephrology program in south-western Ontario. The dialysis treatments compared were hospital hemodialysis, home hemodialysis, self-care hemodialysis, and continuous ambulatory peritoneal dialysis (CAPD). The participants were all patients treated by the same dialysis modality for the fiscal year April 1990 to March 1991. Fully allocated costs are expressed in 1993 Canadian dollars. The average costs per patient year were $88,585 for hospital hemodialysis, $55,593 for self-care hemodialysis, $44,790 for CAPD, and $32,570 for home hemodialysis. The dialysis treatment costs were $54,929 for hospital hemodialysis, $43,313 for self-care hemodialysis, $31,918 for CAPD, and $26,048 for home hemodialysis. These data quantify the magnitude of the differences between fully-allocated costs among the dialysis modalities in a regional nephrology program in Canada. The methodology used in this economic analysis can be applied to programs which differ in structure and scale. The breakdown of dialysis treatment costs into overhead, support department, personnel, supplies, and medication identifies potential areas for cost reduction strategies. PMID- 8714790 TI - Comprehensive typing of HLA-DPB1 alleles by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). AB - Comprehensive typing of 53 HLA-DPB1 alleles was performed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method using 78 polymerase chain reaction-sequence-specific oligonucleotide probe (PCR-SSOP) defined DNA specimens (14 retrospective, 64 prospective). A single primer pair was used to amplify the second exon to obtain DPB1-specific amplified product of 294 bp. A combination of RFLPs and cleaved/uncleaved patterns of various endonucleases was employed to resolve DPB1 alleles. A panel of 13 endonucleases (RsaI, Sau96I, BsrBI, DdeI, BsaJI, BssHII, ScaI, ScaI, BbvI, BsgI, FokI, Bsp1286I and BstUI) yielded unique RFLP patterns for all but 2 pairs of DPB1 alleles. However, these remaining 2 pairs of rare alleles could be resolved by an additional digestion with AciI (DPB1*3901 from 4001 and DPB1*4901 from 5301). The unique RFLP patterns of 21 DPB1 alleles using PCR-SSOP typed DNA specimens had been verified. Of the 1,378 possible heterozygotic patterns, 69 pairs and a triplet had been identified that would yield identical RFLP patterns. However, all but one pair, DPB1*3901/5301 from 4001/4901, of these heterozygotes could be resolved by double digestions with appropriately selected endonucleases from the panel used here. Thus, PCR-RFLP remains a simple and effective method for high resolution DPB1 typing. PMID- 8714791 TI - Hereditary breast and ovarian cancer: epidemiology, genetics, screening and predictive testing. AB - Inherited predisposition to cancer has become an increasingly important part of the practice of clinical genetics. Hereditary breast and ovarian cancer form a significant part of this new field. There are approximately 13,000 new cases of breast cancer diagnosed every year in Canada, and 5% of these can be expected to have a hereditary basis. Large studies implicated a dominant gene with a population frequency of 0.33% and high penetrance. Linkage analysis localised one such gene to chromosome 17q21 in 1990, and 1994 BRCA1 was cloned. The BRCA1 gene is large, and mutations are spread over the entire gene. There is evidence of common origins for some of the mutations. Early detection of hereditary breast and ovarian cancer is problematic. There is no evidence that early diagnosis by population-based mammography improves survival in sporadic premenopausal breast cancer. Multimodal screening for ovarian cancer has not been adequately assessed. Moreover, there is little evidence that such procedures are more likely to be effective in high-risk groups. Women who may gain initial benefit from the cloning of BRCA1 are from BRCA1-linked families who have living affected relatives. Over the next few years, it is likely that predictive testing for hereditary breast and ovarian cancer will become a routine clinical service in large centres. PMID- 8714792 TI - The 1994 G. Malcolm Brown Lecture. Biological barriers and medicine. AB - Career support provides the essential element needed for the detailed development of thematic areas of inquiry. Dr Goresky outlines how career support provided for him the substratum for the investigation and definition of tracer exchange processes in the liver. Classical interstitial substances (labeled albumin, inulin, sucrose, and sodium) and labeled water were found to undergo flow limited distribution in the liver. An excluded volume phenomenon created systematic variation in accessible interstitial space (that is, Disse space) values, smaller for larger molecular weight substances. Labeled rubidium entered liver cells in a concentrative fashion (return from cells was very small, early in time). Labeled glucose liver cell entry was found to exhibit the characteristics of a carrier mediated membrane transport process. Labeled galactose showed saturation of intracellular sequestration, as well as of the cell entry process, the former at much lower galactose concentrations. Straight-chain labeled monohydric alcohols were found to enter liver cells in a flow-limited fashion. Labeled ethanol consumption, when related to underlying steady ethanol values, exhibited Michaelis-Menten kinetics. A substantial extra intracellular enzymic space of distribution was found for the C3- and C4- straight-chain alcohols, predicted by the kinetics. Development of the tracer approach allowed the differentiation and characterization of the processes by which substances penetrate the cell membrane barrier and those resulting in intracellular sequestration. PMID- 8714793 TI - How should excess admission laboratory test utilization be curtailed? - paediatricians' preferences. AB - Routine admission casefinding tests are costly and rarely lead to changes in management. Strategies have been proposed to decrease them. However, it is unclear which tests are considered routine at admission. Our objectives were to clarify which tests are considered to be routine and to determine which methods of reducing test utilization are most acceptable. Sixty-five physicians from a paediatric teaching hospital were selected by random stratified sampling. A questionnaire was administered asking about the role of routine tests and preferences for 6 strategies to reduce test ordering. Sixty percent of respondents felt that more casefinding was occurring than should be, and 89% would agree to a trial of test reduction strategies. Just over half the respondents felt that hemoglobin determination and urinalysis were routine admission tests; fewer physicians listed a wide variety of other tests. Physicians who thought that all admissions should be screened were older, graduated earlier, and tended to be community based. Educational strategies to reduce testing were more acceptable than administrative ones. We conclude that many physicians would support the trial implementation of test reduction strategies in childrens' hospitals. Hospitals wishing to reduce excess admission casefinding should implement educational strategies, as physicians prefer these. PMID- 8714794 TI - The underestimated long term medical and economic consequences of rheumatoid arthritis. AB - Rheumatoid arthritis is generally a progressive disease, with radiographic evidence of joint damage, declines in functional status, and premature mortality. In addition, the disease has severe long term economic consequences, including direct costs of medical care, indirect costs of work disability and interference with social roles, as well as the intangible costs of pain, fatigue, helplessness, loss of self-efficacy, and other psychological difficulties. The consequences of rheumatoid arthritis have often been underestimated by health professionals, reimbursement agencies, the general public, and even rheumatologists. Furthermore, the adverse effects of potential therapies have often been regarded as more severe than the 'adverse effects' of untreated disease. More than 50% of patients with rheumatoid arthritis who are younger than 65 years and who were working at onset of disease receive work disability payments, and 0.8% of the US population eligible to work are individuals who have symmetrical polyarthritis but who, unfortunately, are not working. The total earnings gap between individuals with symmetrical arthritis and those in the general population was $US8.9 billion for women and $US8.7 billion for men, for a total of $US17.6 billion (1986 US dollars). The projected costs of knee replacement in patients with rheumatoid arthritis in the US are between $US600 million and $US900 million annually. New approaches to therapy, which include earlier and more aggressive intervention, new drugs, and combinations of drugs, appear required to provide adequate control of inflammation, so that the long term damage of rheumatoid arthritis might be prevented and the considerable costs ameliorated. The possible adverse effects and costs of treatment must be balanced against the adverse effects and underestimated costs of rheumatoid arthritis. PMID- 8714795 TI - Prognostic markers in rheumatoid arthritis and classification of antirheumatic therapies. AB - Rheumatoid arthritis is characterised by a generally poor outcome and high morbidity, and has a variable course. Identifying those patients most likely to have a poor prognosis is of key clinical significance. Disease outcome can be predicted from a variety of prognostic markers. Some of these are simple demographic features of the patients, and include age, disease duration, and gender. Others are more specific features of rheumatoid arthritis, including the presence of early erosive changes on plain radiographs, high rheumatoid factor titres, high levels of C-reactive protein, and high scores for disease activity. Although no single marker has adequate specificity or sensitivity to form the basis of clinical decisions, the presence of several is predictive of more severe disease. Thus, patients with early erosive damage who are seropositive for rheumatoid factor and have high levels of C-reactive protein are more likely to have a poor outcome. New markers and imaging techniques are likely to become the prognostic tools for the future. These include genetic markers, and a combination of magnetic resonance imaging and dual energy x-ray absorption scans for localised osteoporosis. PMID- 8714796 TI - Clinical use of cyclosporin in rheumatoid arthritis. AB - Rheumatoid arthritis is a chronic immune-mediated disease characterised by an inflammatory synovitis and extra-articular manifestations. There is an expanding body of evidence to indicate that the activation of T lymphocytes is central in the initiation and perpetuation of this disease. Cyclosporin is an immunomodulator and a highly specific inhibitor of T-lymphocyte function, and has demonstrated disease-modifying properties in clinical studies in patients with rheumatoid arthritis. A concern with the use of cyclosporin has been the development of dose-dependent adverse effects, in particular renal dysfunction. Cyclosporin is lipophilic by nature and the conventional oral formulation (Sandimmun) was subject to incomplete and highly variable absorption, resulting in substantial inter- and intrasubject variations in peak concentrations and systemic bioavailability. A microemulsion-based formulation of cyclosporin (Neoral) has recently been developed, and possesses more predictable and improved absorption with a consequent increased peak concentration and systemic bioavailability. The improved predictability of absorption, and hence blood concentrations, facilitates the ability to 'tailor' therapy to an individual patient, which, in theory, could translate into an improved efficacy and safety profile. PMID- 8714797 TI - Combination therapy with cyclosporin in rheumatoid arthritis. AB - During the last few years, there have been rapid developments in the drug treatment of rheumatoid arthritis. This has led to research into the role of combination therapy. While past combinations of slow-acting antirheumatic drugs have resulted in either an excessive number of adverse events or a lack of increased efficacy over single agent therapy, newer combinations appear to be more promising. It has recently been demonstrated that the combination of cyclosporin and methotrexate is more effective than methotrexate alone, with no increase in adverse effects. Improved trial methodology and a better understanding of the mechanism of action of these newer agents are assisting in the development of new combinations. PMID- 8714798 TI - Safety aspects of cyclosporin in rheumatoid arthritis. AB - While cyclosporin has an established role in the management of patients with rheumatoid arthritis, its use can be accompanied by various adverse events. In daily practice, the potential for cyclosporin to induce nephropathy is of most concern and has, to some extent, limited the use of this drug. However, these effects on renal function are dose dependent. Two international meetings have resulted in the publication of guidelines aimed at limiting the risk of adverse events when cyclosporin is used in patients with rheumatoid arthritis. The 1994 International Consensus guidelines with respect to renal safety recommend the principal dosage regimen of 'go low, go slow', which requires starting patients on a daily dose of between 2.5 and 3.5 mg/kg with the maximum daily dose not exceeding 5 mg/kg. The cyclosporin dosage chosen should be titrated against the highest acceptable increase in serum creatinine, i.e. a 30% increase over the baseline value. Renal biopsy data support these guidelines, and when the latter are followed by physicians, it is very unlikely that histological damage will occur. The new oral microemulsion-based formulation of cyclosporin (Neoral) has a more predictable absorption and a 20% greater bioavailability than the conventional formulation (Sandimmun). Therefore, the microemulsion formulation may enable more patients to receive effective treatment at lower doses. This could have interesting safety and pharmacoeconomic implications. PMID- 8714799 TI - International consensus recommendations on cyclosporin use in rheumatoid arthritis. AB - Guidelines for the use of cyclosporin in patients with rheumatoid arthritis have been formulated as a result of 2 International Consensus meetings. Clinical experience since the last meeting in June 1994 indicates that the clear and simple recommended guidelines remain valid. Experience in early identification of patients with a poor prognosis is growing, and trials currently underway will provide further evidence regarding the use of cyclosporin earlier in the course of severe disease. Confirmation of the potential benefits of the microemulsion based formulation of cyclosporin (Neoral) relative to the conventional formulation (Sandimmun) will further strengthen the position of the drug in the management of patients with rheumatoid arthritis. PMID- 8714800 TI - The reliability of depression diagnosis in chronic low back pain. A pilot study. AB - The variability in estimated rates of major depressive disorder (MDD) in chronic pain samples may be accounted for by sample and methodological differences. Most studies rely on a single measure of depression and lack independent or repeated measures. This study investigated the prevalence of psychiatric disorders in a convenience sample of 18 patients disabled by low back pain (LBP) referred to, evaluated, and treated in a comprehensive pain rehabilitation program specifically designed to restore work readiness and return to work. Subjects had one or more spinal conditions, had work-related back pain for more than 6 months (x = 23.8), were Caucasian, predominantly young (x = 37.5) and male (55%), and were disabled on workers' compensation and approved for rehabilitation. The performance of three diagnostic procedures, the Pain Medicine Evaluation, an independent Structured Clinical Interview for DSM-III-R (SCID), and Longitudinal Clinical Diagnosis were compared with the "gold standard" of the Final Diagnosis which incorporated information from the other three. Also, the performance of a questionnaire, The Pain & Mood Structural Interview (PMSI), designed to assess the causal relationship of pain and mood and specifically the confounding effects of physical factors, was examined. Thirteen of 18 patients (72%) were diagnosed with MDD at Final Diagnosis. MDD diagnosis was made on 4 of 18 patients (22%) on the initial Pain Medicine Evaluation and on 9 of 18 patients (50%) on the SCID, with only 1 ruled out by the confounding questionnaire, for a SCID current prevalence of 44%. Both the SCID and Pain Medicine Evaluation accurately diagnosed the MDD cases they identified; 9 of 13 MDD cases (69%) were precipitated by a physical factor other than their pain. Three episodes of MDD were ruled out by confounding. These findings are discussed in relation to their implications for assessing pain patients for psychiatric comorbidity. PMID- 8714801 TI - The efficacy of psychosocial treatments in primary care. A review of randomized clinical trials. AB - Depression and anxiety are the most common psychiatric disorders among ambulatory medical patients and are associated with significant functional disability. However, they remain underrecognized and/or inadequately treated. The AHCPR Depression Guideline Panel recently reviewed the efficacy of pharmacologic interventions for mood disorders in the primary care section, but there are as yet no comparable analyses of the efficacy of psychosocial interventions for primary care patients experiencing depression or anxiety. This review of randomized, controlled trials conducted in primary care settings generally supports the efficacy of psychosocial treatments (PSTs) provided to ambulatory medical patients with psychiatric disorders. However, methodologic deficiencies in these trials (i.e., diagnostic classification of study subjects, attrition patterns, and criteria for assessing treatment response) limit the generalizability of their findings to routine practice. Studies evaluating the effect of PSTs on health care cost and utilization found changes in physicians' prescribing practices, patients' use of psychotropic medication, and number of patient visits to primary care physicians. However, results varied across studies because of methodologic deficiencies similar to those noted previously. Suggestions are offered for improving the internal and external validity of randomized PST trials in primary care settings. PMID- 8714802 TI - Detecting psychological distress among patients attending secondary health care clinics. Self-report and physician rating. AB - A study was conducted to determine the prevalence of psychological distress, as reported by patients and their physicians, in orthopedic, neurology, dermatology, and ophthalmology clinics; to study their accuracy in detecting psychological distress; and to determine if there is any connection among psychological distress, accuracy of detecting distress, and use of mental health and primary health care physicians' prognosis for the somatic complaints. Five hundred and fifty-six patients, ages 18-21, responded to the Psychiatric Epidemiology Research Interview Demoralization Scale (PERI-D), a measure of psychological distress, and to questions about their mental health and use of mental health and primary health services. Physicians, who were blind to patients' responses, were asked to what extent they thought the cause of patients' complaints was physical and to what extent they thought it was psychological in nature, and to prognosticate. Based on the PERI-D, about 25% of patients were distressed, this was less for females than males and varied between clinics. Based on self reporting, about 14% of patients (males and females) were distressed. Based on physician reporting, about 17% (males less) were distressed. Physicians identified 35% of the PERI-D-distressed cases and 79% of nondistressed cases. About 66% of patients identified their distress and 83% their lack of distress. Increased use of primary health care and mental health care was related to distress. The prognosis was negatively related to distress. Based on this study, there is a need for more attention to psychological distress among secondary health care patients. Patients' ability to identify their distress suggests the importance of involving the patient in the diagnostic process. Correct detection of distress alone does not appear to decrease the use of primary medical and mental health services. PMID- 8714803 TI - Recent life events, gender differences, and the control of insulin-dependent diabetes mellitus. A 2-year follow-up study. AB - In an earlier study of patients with insulin-dependent diabetes mellitus (IDDM), males who reported predominantly negative life events over the previous year and had a poorer social support situation showed poorer HbA1C values than those who reported fewer or no negative life events. For the females it was found that the greater the number of life events reported, especially positive ones, the greater the change for the better was HbA1C over the event year studied. The present study aimed at following up, during the next event year period, various gender specific patterns obtained in the previous study. For the males, negative life events and HbA1C values were found to be positively related this second event year as well. In addition, more negative life events were reported by those males who, in the previous study, were defined as high-negative eventers. In contrast, for the females, no significant correlations were obtained between life events and HbA1C values for the second event year. The results are discussed in terms of possible differences in psychosocial environment and coping strategies between males and females. PMID- 8714804 TI - Psychiatric evaluation of physical rehabilitation patients. AB - We conducted a study to demonstrate the frequency and types of psychiatric/psychological symptoms. A Structured Interview according to the DSM III-R was conducted which demonstrated that 46 (41.4%) of 111 rehabilitation inpatients met the criteria for some forms of psychiatric disorders: 34 patients for major depression, 10 for adjustment disorder with anxious mood, and 2 for posttraumatic stress disorder. The remaining 65 patients (58.6%) showed normal reactions to their diseases. Average length of hospital stay for patients with major depression was significantly longer than those with no or the other types of psychiatric disease. They were also tested with Zung's Self-Rating Anxiety Scale (SAS), Zung's Self-Rating Depression Scale (SDS), and Profile of Mood States (POMS). Three psychological tests were useful in detecting depression or adjustment disorder among rehabilitation patients; however, these tests are not always specific to the type of psychiatric disorders. Patients with higher scores in those three tests should be referred to a psychiatric consultant for detailed examinations and proper treatments, if necessary. PMID- 8714805 TI - Treating the patient who is disfigured by head and neck cancer. AB - A 46-year-old man with laryngeal carcinoma was admitted to the medical service for lethargy. The medical team requested a psychiatric consultation to assist with the patient's depression, substance abuse, and noncompliance. The case is presented and discussed with reference to the issues of depression, disfigurement, dysfunction, and substance abuse in the patient with head and neck cancer. PMID- 8714806 TI - Patients' threats. Expanded definition of assault. AB - Most studies of patient assaults against staff operationally define violence as episodes of unwanted physical or sexual contact. This study empirically assessed a broader range of patient violence by including verbal and nonverbal threats in addition to sexual and physical assaults. Preliminary data from a statewide survey suggested that patient threats were frequent events. Data from a team of staff assaulted by patients in one hospital suggested that some verbal threats produced as much psychological distress for staff victims as did some physical assaults. These findings suggest the need to consider including threats in future studies of patient assaults. PMID- 8714807 TI - The cost of antidepressant overdose. AB - Ninety percent of suicide attempts referred to a general hospital are by self poisoning. Among women, drug overdose is the commonest means of suicide. In a retrospective naturalistic review of 200 patients who were treated in the Critical Care Unit of a general hospital following medication overdose, 12% were antidepressant overdoses. The mean duration of hospital stay for overdose with tricyclic antidepressants (TCA) was more than double that for overdose with selective serotonin reuptake inhibitors (SSRI) (7 vs 3 days; z = 2.20, p < 0.05). The dollar cost of hospital treatment for patients who overdosed on TCAs was four times greater than that for patients who overdosed on SSRIs ($22,923 vs $5,379; z = 2.30, p < 0.05). The tricyclic compounds clearly have a price advantage over more recently introduced antidepressant agents fluoxetine, sertraline, paroxetine, venlafaxine, and bupropion. The apparent cost advantage of prescribing a less expensive drug may be nullified by the cost associated with adverse consequences. PMID- 8714808 TI - ECT-valproic acid interaction. PMID- 8714809 TI - Psychotic episode related to phenylpropanolamine and amantadine in a healthy female. PMID- 8714810 TI - Hyponatremia and Stevens-Johnson syndrome in a patient receiving carbamazepine. PMID- 8714811 TI - Acute confusion in a chronic benzodiazepine patient. Withdrawal-related nonconvulsive status epilepticus misdiagnosed as acute intoxication. PMID- 8714812 TI - Microvascular architecture and exchange in teeth. AB - Dental pulp is a low-compliance tissue surrounded by an avascular hard tissue case. Resin casts of the microvasculature in teeth of limited growth show arterioles and venules arranged axially in the pulp with capillary loops extending out toward the dentine. The capillary density is high. There is ultrastructural evidence for lymphatics in pulp. Pulpal blood flow has been estimated in intact teeth using radiolabeled microspheres and found to be in the range 20-60 ml/min per 100 g tissue. One of the difficulties of studying this tissue is that a cavity has to be cut into the tooth with the inherent risks that this may affect the parameters under study. Measurements from exposed pulp indicate that the tissue fluid pressure is high and pulsatile. Furthermore, micropuncture studies have shown that the arteriolar pressure is lower and the venular pressure higher than in other tissues. When dentine is exposed in vivo, fluid moves out through the dentinal tubules and this appears to be formed by a process of ultrafiltration from the pulpal interstitial fluid. The flow is sufficient to retard significantly the diffusion of chemicals into dentine from the oral cavity. PMID- 8714813 TI - Early in reperfusion following myocardial ischemia, leukocyte activation is necessary for venular adhesion but not capillary retention. AB - OBJECTIVE: The pathobiology of leukocyte sequestration in the coronary microcirculation following ischemia is unclear. We examined the location(s) and persistence of leukocyte sequestration of unactivated and preactivated blood in the coronary microcirculation early during reperfusion following ischemia. METHODS: Isolated rat hearts were subjected to 30 min of 37 degrees C, no-flow ischemia. Hearts were initially reperfused with diluted whole blood containing fluorescent leukocytes (DWB*). At 5, 20, and 35 min of reperfusion (R), the deposition of leukocytes in the coronary capillaries and venules was observed directly using intravital fluorescence microscopy. Four groups were studied: a nonischemic control group (Gr I), and postischemic groups reperfused with DWB* treated with vehicle (Gr II) or preactivated with 10(-8) M N-formylmethionyl leucyl-phenylalanine (fMLP) (Gr III) or 10(-6) M fMLP (Gr IV). RESULTS: At R5, postischemic reperfusion with unactivated blood caused a significant trapping of leukocytes in coronary capillaries (Gr I = 2.2 +/- 0.4 versus Gr II = 5.6 +/- 0.6 leukocytes per capillary field, P < 0.05). Hearts in Gr IV exhibited significantly greater leukocyte retention in capillaries compared to all other groups at R5 (R5, Gr IV = 8.8 leukocytes per capillary field, P < 0.05) and at R35. At R5, although more leukocytes were observed adhered to the venules in Gr II compared to Gr I, the difference was not statistically significant (Gr I = 1.7 +/- 0.7 versus Gr II = 3.4 +/- 0.5 leukocytes per 100 microns venule, P = 0.23). DWB* preactivated with the lower concentration of fMLP (10(-8) M) resulted in a significant increase in venular leukocyte adhesion at R5 compared to Gr I and Gr II (Gr III 6.1 +/- 0.5, P < 0.05). After 35 min of reperfusion, a greater percentage of leukocytes remained in the capillaries than in the venules. CONCLUSIONS: These direct observations suggest that early in reperfusion after ischemia, both leukocyte and endothelial activation are necessary for venular adhesion, but that ischemia-induced coronary microvascular alterations are sufficient to promote leukocyte retention in coronary capillaries. These results also indicate that during 35 min of reperfusion, the degree of leukocyte washout is greater in the venules than in the capillaries. These results suggest that the mechanisms contributing to leukocyte retention early in reperfusion following myocardial ischemia are, indeed, different in the capillaries and venules and that the mechanisms affecting retention in capillaries are more persistent than those in the venules. PMID- 8714814 TI - Blood vessel growth in the endometrium. AB - Angiogenesis, or formation of new blood vessels by sprout formation from existing vessels, is generally considered to be the only mechanism by which blood vessel growth occurs. This traditional concept of angiogenesis has been derived largely from observations of experimental systems. Relatively fewer studies on angiogenesis have been carried out using normal angiogenic situations where vessel growth occurs in a controlled three-dimensional fashion throughout the tissue. Recent advances in the treatment of infertility and outpatient gynecological procedures have led to greater accessibility to normal human endometrium, thus providing new opportunities to study the process of angiogenesis in a physiological context. However, to date, it appears that very little work had been done in relation to endometrial angiogenesis apart from the location of numerous angiogenic and other growth factors with potential to influence angiogenesis in the endometrium, and here there have been few attempts to link these observations with actual angiogenic events. The purpose of this review is to summarize the literature regarding angiogenesis in the endometrium, including work from our own laboratory, and to suggest that blood vessel growth in the endometrium may occur by a mechanism that differs from classical angiogenesis. PMID- 8714815 TI - Plasma skimming in vascular trees: numerical estimates of symmetry recovery lengths. AB - A new model of plasma skimming in vascular trees that includes the history of disturbances at previous bifurcations is developed. The model calculates the shift in the red blood cell concentration profile by mapping streamlines through the junction; it then computes the dissipation of that disturbance through random collisions among the cells. A numerical solution of this model is compared to experimental data on plasma skimming in serial bifurcations. Numerical values of symmetry recovery lengths downstream of the bifurcation are calculated using the model. Comparison of the computed symmetry recovery lengths with anatomical measurements in hamster cremaster muscle indicate that asymmetries in red blood cell concentration profiles are important in vessels with diameters below 55 microns. PMID- 8714816 TI - Responses of cremasteric arterioles of spontaneously hypertensive rats to changes in extracellular K+ concentration. AB - OBJECTIVE: The goal of this study was to determine the effect of changes in extracellular K+ concentration ([K+]o) on active tone in cremasteric arterioles of spontaneously hypertensive rats (SHR) and their normotensive Wistar-Kyoto (WKY) and Wistar controls. METHODS: Diameters of third- and fourth-order arterioles were measured in the cremaster muscle of hypertensive and normotensive rats during abrupt changes in superfusate K+ concentration from 4.7 mM to 0 mM to 15 mM K+. RESULTS: Arterioles constricted in response to superfusion with 0 mM K+ and exhibited a large, transient dilation in response to an abrupt change from 0 mM to 15 mM [K+]o. Arteriolar dilation in response to 15 mM K+ was significantly larger in 12-15-week-old SHR than in WKY or Wistar controls. Arteriolar responses to 15 mM K+ were not significantly different in 4-6-week-old SHR and WKY. Dilator responses to 15 mM K+ were generally inhibited by 1 mM ouabain, although ouabain was less effective in inhibiting 15 mM K(+)-induced dilation in arterioles of SHR and WKY than in Wistar rats. CONCLUSIONS: Dilation of cremasteric arterioles in response to 15 mM [K+]o is mediated, at least in part, via stimulation of the electrogenic Na+/K+ pump, although Na+/K(+)-pump-independent components may also contribute to the response. Arterioles of SHR with established hypertension exhibit an altered response to elevated [K+]o which is not present in SHR in the early stage of hypertension. PMID- 8714817 TI - Use of Ruthenium Red staining to detect mast cell degranulation in vivo. AB - OBJECTIVE: To establish a method of detecting mast cell degranulation in tissues during in vivo microscopy. METHODS: Hamster tissues were prepared for intravital microscopy. Ruthenium red (RR) was superfused over the cheek pouch at concentrations of 0.0001-0.01% to determine the optimal concentration. Mast cells were stimulated with compound 48/80, as well as with vasoactive agents not known to be stimulatory to mast cells, following which, mast cell staining was observed. Mesenteries were stained with Toluidine Blue (TB) or RR and mast cell degranulation was assessed during treatment with compound 48/80, or control. RESULTS: During superfusion with varying concentrations of RR, a dose dependence for background staining of unstimulated cells was observed. A RR concentration of 0.001% was optimal for in vivo detection of mast cell degranulation. Mast cells exposed to 0.001% RR were stained following stimulation with compound 48/80 but not after treatment with KCl or acetylcholine. The latter agents are not known to stimulate mast cells. Thus, arteriolar vasomotor responses, per se, did not appear to play a role in mast cell RR uptake. Comparable results were obtained with RR versus TB in control or 48/80-treated mesenteries. CONCLUSIONS: This RR technique facilitates rapid detection of mast cell degranulation in vivo and provides an opportunity to assess both mast cell and microvascular function simultaneously. PMID- 8714818 TI - A role for dietary copper in nitric oxide-mediated vasodilation. AB - OBJECTIVE: This study was designed to investigate the role of dietary copper in nitric oxide-mediated arteriolar dilation. METHODS: Male weanling Sprague-Dawley rats were fed a purified diet that was either copper-adequate (6.0 micrograms Cu per g diet) or copper-deficient (0.3 microgram Cu per g diet) for a period of 4 weeks. Each rat was anesthetized with pentobarbital and its cremaster muscle was positioned in a Krebs'-filled bath to which graded concentrations of vasoactive agents were added. In the first series, responses to norepinephrine (NE 10(-9) 10(-6) M) and acetylcholine (ACH 10(-7)-10(-4) M) were compared in third-order arterioles. Second, the dilator response to 10(-5) M ACH in the absence and presence of 240 U/ml Cu, Zn-superoxide dismutase (SOD) was determined. Third, arteriolar dilation was determined in response to NO-independent stimulation of soluble guanylate cyclase with hydrogen peroxide (10(-7)-10(-5) M) and to dibutyryl cGMP (10(-6)-10(-4) M), dibutyryl cAMP (10(-6)-10(-4) M), and papaverine (10(-4) M). RESULTS: The arteriole constrictor response to NE and the dilator response to hydrogen peroxide, dibutyryl cGMP and cAMP, and papaverine were not different between the dietary groups. Copper deficiency attenuated the ACH-induced dilation, but the response was restored in the presence of SOD. CONCLUSIONS: The inactivation of cytosolic Cu, Zn-SOD by restriction of dietary copper results in the depression of NO-mediated vascular smooth-muscle relaxation probably by interaction of NO with superoxide. PMID- 8714819 TI - A system for culture of endothelial cells in 20-50-microns branching tubes. AB - OBJECTIVE: To construct an in vitro endothelial cell culture system which would mimic the geometry and hemodynamic conditions of the arteriolar microcirculation. METHODS: Using a photolithography technique, semicircular channels (20-50 microns in diameter) were etched in mirror-image patterns on pairs of borosilicate microscope slide glass. One-half of each plate pair was predrilled with perfusion port holes at funnel-shaped fluid entrance regions. The perfusion system was constructed of micropipette glass and Teflon tubing, and imbedded in Sylgard. Two types of endothelial cells were grown to confluence within the half-channels: rabbit lung microvascular endothelial cells (a gift of Dr. M.E. Gerritsen, Miles Inc.) and human umbilical vein endothelial cells. After the cells were confluent, the two mirror images were aligned and clamped together to form a complete branching system of tubes lined with endothelial cells. RESULTS: This cell culture system can be perfused at physiological flow rates corresponding to wall shear stress values in the range 0.03-48 dyn/cm2. The fluid velocity profiles can be measured in this system by tracking the velocity and flow paths of 0.5-micron fluorescently labeled microspheres. Endothelial cells which grow within the channel exhibit F-actin alignment along the long axis of the channel by 3 days after seeding. Scanning electron micrographs indicate that 4 hr after seeding, endothelial cells commonly form cellular projections extending across the half channel; by 5 days after seeding, the projections appear to have flattened out along the bottom of the channel. CONCLUSIONS: An in vitro endothelial cell culture system was constructed which mimics the geometry and hemodynamics conditions of resistance arterioles. This system can be used to examine endothelial cell responses to flow and flow gradients under defined and controllable conditions which mimic the arteriolar microcirculation. PMID- 8714820 TI - [Current concepts of the interactions of regulating systems: nervous, endocrine and immune]. AB - Three regulating systems, nervous, endocrine and immune, are involved in maintenance of homeostasis. They are regarded as interacting, with interaction between nervous and endocrine systems being well defined and giving rise to development of independent realm of knowledge--the neuroendocrinology. As concerns interaction between the neuroendocrine and immune systems it seems to be exciting and intensely developing trend of up-to-date investigation. A great deal of evidence is adduced bringing new sight into the mechanisms of bidirectional exchange of signals among the nervous, endocrine and immune systems. This paper provides analytical consideration of the vast information, that allowed the development of the new integrative area of biology--the neuro-immuno endocrinology, with special reference being paid to some general regularities underlying integration of each regulatory constituents into the single operating system. Attention is also paid to up-date information concerning mechanisms of neurotransmition and immune response in terms of the problem under consideration. PMID- 8714821 TI - [Neurospecific protein-hormonal complexes]. AB - New cardioactive protein-hormone complexes (PHC) were identified, and then separated and purified in magnocellular foci of hypothalamus of oxen and some other animals. It was proved that the PHC are specific for nervous tissue, their subsell localisation was studied, concentration and distribution in an organism were identified. It was discovered, that under dissociation of PHC, its high molecular "apoforms" represent new specific glicoproteins, and the low-molecular combinations related to them in a non-covalent way are glicopeptides with clear cardioactive properties inherent in the respective native PHC. The above glicoproteins are transport molecules for the cardioactive neurohormones of hypothalamus, and are the basis for mediation of the inter-neurone interaction and the finction of intracellular connection. In addition, it was proved that these glicoproteins can exist in the form of preprohormone forms of cardiotropic neurohormones of hypothalamus: "K", "S", and "G". PMID- 8714822 TI - [The peptidergic correction of hemorrhagic shock]. AB - The study of haemorrhagic shock mechanisms and methods of its correction is of great special theoretical and practical interest. The authors described the new data about the action of opioid agonists and antagonists by the haemorrhagic shock with the special attention of the effects of paraopioid peptide FMRFa. The new field of scientific interests in the use of peptide mixture in the subthreshold doses by the haemorrhagic shock correction and the influence of opioids and its antagonists on the neurologic status in the posthaemorrhagic period. PMID- 8714823 TI - [The thyroid gland as a modulator of ovarian development and histophysiology]. AB - On the base of the complex of experiments modeling disfunctional states of the thyroid gland were proved the most importance of the thyroid gland hormones in regulation of development and histophysiology of the ovary. There were analyzed possible mechanizmes by with thyroid gland hormones influence on this biological processes and its role in general system of the neurohumoral regulation of sexual function. PMID- 8714824 TI - [Evoked potentials: their spatial physiological characteristics and the problem of their functional assessment]. AB - Basic data about physiological evoked responses properties of the brain structures highest level (telencephalon) are presented. Contemporary ideas concerned to mechanisms of these responses are summed up. Data about their distribution in telencephalic structures and morphological basis of the distribution are adduced. The evoked responses especialities of telencephalic structures in awake animals in chronic experiments are revealed. The functional significance of processes that form this bioelectrical reaction is observed. Special attention is paid to earlier (to 100 ms) components. The possibility to use the evoked responses as the functional instruments of investigations of the brain is discussed. PMID- 8714825 TI - [The individual manifestations of respiratory rhythms]. AB - The information about the individual peculiarities of the respiratory rhythmics of health people is produced in this article. It is shown, that the functional tests: voluntary hyperventilation, hypoxic hypoxia, their combined influence and natural sleep in normal and hypoxic conditions may be the convenient models for researching this problem. The dependence of the respiratory responses on the individual type of respiration in normal conditions is revealed. The authors concluded that not only the character of normal breathing but also the reactions on functional tests should be taken into account during the prognosis of alterations of the respiratory rhythmics in various conditions. PMID- 8714826 TI - [Behavioral activity in an uncertain environment (the methodological characteristics of an experimental study)]. AB - A fundamental resemblance between processes of probabilitive training and conditional reflex forming exist wile using lowintensive signal stimulus and probabilitive way of confirmation. A base of intrinsic unity of these methodical variants is a complicated way of elicitation of bond between signal stimulus and confirmation, absence of one-way interpreted information about an environment presuming probabilitive estimate using. In this case informative interaction between an individual and subjectively accidental environment takes place or the same, a process of subjectively probabilitive training. Accordingly probabilitive training is a dinamic process of training under conditions of subjective uncertainty based probabilitive estimate application and feedback elements existence. PMID- 8714827 TI - [A shift in types of nutrition and digestion in ontogeny]. AB - The article generalises research data from experiments with animals and observations on man. These allow to identify specific features of digestion, as well as of alimentation starting from a zygote stage, which are organised and developed by the organism itself. The following stages were outlined: hystotrophic stage--that of an embryo; hematrophic, which is combined in the foetal stage with the omntrophic and lactrophic after birth. The article discusses the extreme importance of the latter, both in the trophogenic aspect, as well as for determining the future post-natal formation of the organism advanced in the biological, as well as in the socio-economic aspects. PMID- 8714828 TI - [Nikolai Nikolaevich Sirotinin and his concept of the evolution of body resistance and reactivity (on the centenary of his birth)]. PMID- 8714829 TI - [Neurosecretion--a fundamental pattern of neurobiology (the results of the 12th International Symposium Neurosecretion-95, Germany, Kiel, 19-23 September 1995)]. PMID- 8714830 TI - [Results and main tasks of the activities of this journal]. PMID- 8714831 TI - [The functional characteristics of the cardiovascular system in children exposed to air pollution]. AB - Clinicophysiological survey of the children living in polluted ambient air areas has revealed the high incidence of cardiovascular functional, adaptative, and regulatory disorders at rest and after graded exercise. PMID- 8714832 TI - [The hygienic characteristics of drinking water consumption in ecologically unhealthy regions]. PMID- 8714833 TI - [The hygienic assessment of a new technology for decontaminating electroplating wastes]. PMID- 8714834 TI - [Hygienic assessment of the quality of drinking water conditioned by low-voltage pulsed electrical discharges]. AB - The purpose of this study was to make experimental and semifield examinations of the physical and chemical indices of the quality of water disinfected and preserved by low-voltage (2.8-3.0 kV) impulsive electric discharges by means of a MEI portable plant. The findings showed that the changes in the physical, chemical, and organoleptic properties of the treated water were insignificant. The water kept during 2 months met the requirements for drinking water. The above portable plant is recommended for water disinfection in self-contained objects. PMID- 8714835 TI - [Characteristics of the progression of silicosis under alpine conditions]. AB - Siliceous dust during hypobaric hypoxia under the conditions of high mountains causes a rapid onset of silicosis in miners. Nodular silicosis types are prevalent. Lower activity of antioxidative protective enzymes and activated free radical oxidation and autoimmune responses are detectable. PMID- 8714836 TI - [The causes, diagnosis and treatment of iron-deficiency states in the population (a review)]. AB - Investigation has indicated that the main cause of blood iron deficiency is insufficient dietary intake of iron, vitamins, and proteins. The comprehensive examination of all iron metabolism-associated indices may identify iron deficiency, hematological parameteres being much more preferred. Iron drugs in combination with other trace elements, vitamins, and amino acids should be prescribed in this abnormality. PMID- 8714837 TI - [Population monitoring of the physical development of the pediatric population]. PMID- 8714838 TI - [The effect of the life style on the physical development and health status of schoolchildren]. AB - Questionnaires were used in children to study risk factors (diets, motor activity, bad habits, awareness of their own health status) for non-infectious diseases. Conclusions on the impact of lifestyle on their physical development and development of abnormalities were made. PMID- 8714839 TI - [The personality characteristics of today's adolescents and means for their correction]. AB - Experimental design for organizing the activity of 14-17-year-old adolescents was proposed. The design includes various exercises, games, visual promotion, etc. The rational organization of leisure time will make goal-oriented correction of adolescents' personal traits and exert a positive effect on the processes of their formation. PMID- 8714841 TI - [The methodological aspects in the ecological hygienic assessment of modern building and finishing materials]. PMID- 8714840 TI - [The assessment of the effect of radiation on the body adaptation potentials in people living on the Techa River]. AB - The functional change index was calculated to assess the strain of adaptative mechanisms in inhabitants on the Techa river polluted with radioactive waste. Radiation was shown to have an impact on the strain of adaptative mechanisms, but the degree of this impact was moderate. PMID- 8714842 TI - [Hygienic characteristics of the state of the environment and of the health of the population in areas with a developed petrochemical industry]. PMID- 8714843 TI - [The experimental evaluation of the potential blastomogenic properties of substances and materials used in medicine]. PMID- 8714844 TI - [The use of the local press for informing the population on health and ecology]. PMID- 8714845 TI - [Characteristics of the hygienic habits of students during an academic course on health education]. PMID- 8714846 TI - [The problem of controlling smoking among schoolchildren]. PMID- 8714847 TI - [A chromatographic method for determining diphenylolpropane in the air of a work area]. PMID- 8714848 TI - [Gas chromatographic determination of ethylene oxide residue in sterilized medical items]. PMID- 8714849 TI - [A uniform quantitative measure of human exposure to harmful substances in ecological calculations]. PMID- 8714850 TI - Similarity in standards and safety of practice within Europe: reality or fantasy? PMID- 8714851 TI - Malignant hyperthermia suggestive hypermetabolic syndrome at emergence from anesthesia. PMID- 8714852 TI - The effect of calcium channel antagonist administered by iontophoresis on the pain threshold. AB - Three calcium (Ca) channel antagonists were administered to healthy adult volunteers by iontophoresis alone or in combination with lidocaine to study their effects on the pain threshold. Nicardipine, verapamil or diltiazem administered alone elevated the pain threshold to the same degree as lidocaine. When Ca channel antagonists were administered in combination with lidocaine, the pain threshold elevation was not changed, but the duration of the analgesic action was prolonged when compared with lidocaine alone. These results suggest a role of the calcium channel in the pain control mechanism and the possible clinical application of Ca channel antagonists delivered by iontophoresis in pain management. PMID- 8714853 TI - EMLA prevents pain during local anesthesia for cataract surgery. AB - In an observer blind study, the efficacy of lignocaine-prilocaine eutectic mixture was established in alleviating the pain of needle insertion during retrobulbar and periorbital tissues block in cataract surgery. Thirty-three patients were allocated randomly into two groups. Patients in group 1 (n = 17) received EMLA cream 60 min. prior to local anesthesia while those of group 2 (n = 16) were used as a control group. Pain was graded by the patients on a four-point verbal rating scale (VRS) as well as by an observer, unaware of the treatment patients received, on a 4-point scale. In both groups there were no differences in pain scores between patient's and observer's assessments. In EMLA group significantly (p < 0.05) less pain was registered during performance of the local anesthetic procedure. Our results demonstrate that the eutectic mixture may be very effective in reducing pain associated with locoregional procedures for eye surgery. PMID- 8714854 TI - Performance of the anesthesiologists in 12 Departments of Anesthesia in Greece. AB - To assess the performance of anesthesiologists in 12 Departments of Anesthesia a questionnaire was completed by 132 anesthesiologists. The questionnaire included preoperative visit, preoxygenation, the anaesthetic record, time of stay in the operating room, management of apnea at the end of surgery and rapid sequence induction. Anesthesiologists were also asked if they keep update and how. Eighty nine percent of the anesthesiologists visit the patients preoperatively, 77% preoxygenate them routinely, 22% preoxygenate only the emergency patients, 0.8% never preoxygenate, 83% keep an anesthetic record, and the average time they stay in the operating room is 84% of the procedures. If the patient is apneic at the end of surgery, 43% of the anesthesiologists use peripheral nerve stimulator, 41% give atropine/neostigmine, 13% repeat the neostigmine, and 82% continue mechanical ventilation. The 12 Departments of Anesthesia differed significantly between them in all the above variables (P < 0.01-P < 0.0001 for each of the variables between them). To induce anesthesia in patients with full stomach, 89% give thiopentone and succinylcholine, 4% thiopentone and nondepolarizing relaxant, 76% apply cricoid pressure and 5% inflate the lungs via a face mask, (P < 0.0001, P = 0.018 and P < 0.0001 respectively). Eighty-seven percent of the anesthesiologists keep update attending congresses, 76% departmental meetings (P = 0.0015 within the 12 Hospitals), 86% reading journals and 78% reading textbooks. The 12 Departments of Anesthesia presented a uniformity in updating knowledge but not in practicing anesthesia. PMID- 8714855 TI - Continuous cervical intrathecal administration of morphine with a new infusion pump, the Anschutz IP 35.1: a case report. AB - The continuous intrathecal administration of low-dose morphine is an effective treatment for chronic neuropathic pain. Several implantable devices can be used for such treatments but they all have limitations. In this case report we opted for a new type of implantable pump, the Anschutz, for the combined cervical intrathecal administration of morphine and clonidine in a patient with severe cervicobrachialgia. PMID- 8714856 TI - Malignant hyperthermia suggestive hypermetabolic syndrome at emergence from anesthesia. AB - A 6-year old female child received succinylcholine (1 mg.kg-1) and isoflurane (concentrations of 1.5-2 percent) and developed at the end of surgery a hypermetabolic syndrome suggestive of malignant hyperthermia (MH) with masseter muscle spasm, muscle rigidity, tachypnea, systolic hypertension (140 mm Hg), tachycardia (205 beats.min-1), hypercarbia (end expiratory CO2 71 mmHg), and an increase in body temperature (39.2 degrees C). The child responded well to therapy which included cooling, hyperventilation with pure oxygen and dantrolene administration. However, blood creatine kinase and myoglobin elevations were moderate (respectively 375 IU.L-1 and 114 micrograms.L-1) and an in vitro halothane and caffeine contracture test was negative. Differential diagnostic proposals are discussed and compared to the clinical incident. PMID- 8714857 TI - Mild pre- and posttraumatic hypothermia attenuates blood-brain barrier damage following controlled cortical impact injury in the rat. AB - Recent studies have demonstrated a neuroprotective effect of mild/moderate hypothermia in models of cerebral trauma and ischemia. In contrast, hypotension is known to exacerbate CNS injury. To better understand the mechanisms whereby hypothermia and hypotension influence secondary neural injury, the present study assessed the effects of these two variables upon blood-brain barrier (BBB) permeability following controlled cortical impact injury. Rats were subjected to either 0, 15, or 30 min of hypotension under normothermic or slightly hypothermic brain temperature conditions. Brain temperature was maintained within 0.5 degrees C of baseline (normothermic) or allowed to float freely (e.g., become hypothermic) throughout the study. Hypotension was induced immediately after head injury by rapid hemorrhage down to a mean arterial pressure of 50 mm Hg and held there for 15 or 30 min. Blood-brain barrier permeability was measured by the extravasation of plasma protein-bound Evan's blue dye into the injured cortex at 60 min postinjury. The results revealed that mild hypothermia (< 1.6 +/- 0.2 degrees C), right before and 15-30 min following head injury, significantly reduced BBB permeability 28.0, 21.8, and 26.2% in rats subjected to 0, 15, or 30 min hypotension, respectively (all p values < or = 0.05). Hypotension did not increase BBB permeability nor did it significantly interact with the brain temperature effect. Previous results, using this same model, have shown that the progressive posttraumatic increase in BBB permeability is preceded by an increase in cortical .OH and lipid hydroperoxides at the site of injury and is attenuated by the lipid peroxidation inhibitor tirilazad mesylate. Thus, the present results are discussed in terms of the role of free radical-induced lipid peroxidation in the genesis of posttraumatic BBB damage and the possible effects of hypothermia upon this injury process. PMID- 8714858 TI - Reduction of the neurological deficit in mice with traumatic brain injury by nitric oxide synthase inhibitors. AB - This study investigates the effect of the NO synthase inhibitors, NG-nitro L arginine methyl ester (L-NAME) and 7-nitro indazole (7-NI), on the neurological deficit 24 h after a moderate closed head injury in mice. Low doses of L-NAME or 7-NI given soon after the injury significantly reduced the neurological deficit compared to the vehicle-treated group. L-Arginine (300 mg/kg) did not alter the neurological deficit, but reversed the protective effects of both L-NAME and 7-NI when given at the same time. Both L-NAME and 7-NI had dose-related effects. The neuroprotective effects of L-NAME and 7-NI occurred when the drugs were given 5, 30, or 60 min after brain injury, but not when treatment was begun 2 h after brain injury, suggesting a short therapeutic window for both drugs. These results suggest that NO synthesis by neuronal NO synthase plays an important role in the early neurotoxic cascade leading to neurological deficit following traumatic brain injury. PMID- 8714859 TI - Thresholds for cerebral ischemia after severe head injury: relationship with late CT findings and outcome. AB - Cerebral ischemic insults in at least 30% of severely head injured patients at a very early stage following trauma and are associated with early death. To date, the threshold for ischemia of 18 mL/100g/min used in human head injury studies has been adopted from animal studies (by temporary occlusion of the middle cerebral artery). Since the traumatized brain becomes more susceptible to irreversible damage if accompanied by ischemia one may question whether the threshold for ischemic vulnerability is higher than 18 mL/100 g/min. Cerebral ischemia can cause atrophy. Therefore, the authors obtained computerized tomography (CT) scans in 33 comatose head-injured patients (Glasgow Coma Score of 8 or less) at least 3 months following injury and compared ventricle sizes (as a reflection of atrophy) with cerebral blood flow (CBF) obtained within 4 h (average 2.3 +/- 0.8 h) after injury. Ventricular measurements were performed in three fashions: the third ventricular size (cm), the bicaudate cerebral ventricular index (BCVI), and the hemispheric ventricular index (HCVI). No significant correlation was found between early CBF and any of the ventricule sizes. Applying a multiple correlation analysis with four independent parameters [CBF, CBF/time postinjury, CBF/(time postinjury)2, age], only age emerged as a significant indicator for predicting ventricle size (p < 0.001). We also compared CBF data, obtained within 4 h after trauma, from survivors at 3 months after injury (mean CBF of 32 mL/100 g/min) with CBF data from non-survivors (CBF 20 mL/100 g/min). The difference in CBF between survivors and nonsurvivors was significant at p < 0.001 (Wilcoxon rank-sum test). The proportion of patients with CBF less than or equal to 20 mL/100 g/min was 56% in the nonsurvivors and only 5% in survivors. The difference in the proportions was significant at p < 0.001 (chi-square test). We conclude that a measure of atrophy does not correlate with ultra-early CBF. However, based on the clear distinction between survivors and nonsurvivors, we suggest the threshold for ischemia after head injury be redefined as a CBF of 20 mL/100 g/min. PMID- 8714860 TI - Zinc supplementation is associated with improved neurologic recovery rate and visceral protein levels of patients with severe closed head injury. AB - Sixty-eight patients were entered into a randomized, prospective, double-blinded controlled trial of supplemental zinc versus standard zinc therapy to study the effects of zinc supplementation on neurologic recovery and nutritional/metabolic status after severe closed head injury. One month after injury, the mortality rates in the standard zinc group and the zinc-supplemented group were 26 and 12%, respectively. Glasgow Coma Scale (GCS) scores of the zinc-supplemented group exceeded the adjusted mean GCS score of the standard group at day 28 (p = 0.03). Mean motor GCS score levels of the zinc-supplemented group were significantly higher on days 15 and 21 than those of the control group (p = 0.005, p = 0.02). This trend continued on day 28 of the study (p = 0.09). The groups did not differ in serum zinc concentration, weight, energy expenditure, or total urinary nitrogen excretion after hospital admission. Mean 24-h urine zinc levels were significantly higher in the zinc-supplemented group at days 2 (p = 0.0001) and 10 (p = 0.01) after injury. Mean serum prealbumin concentrations were significantly higher in the zinc-supplemented group (p = 0.003) at 3 weeks after injury. A similar pattern was found for mean serum retinol binding protein level (p = 0.01). A significantly larger number of patients in the standard zinc group had craniotomies for evacuation of hematoma; thus a bias may have been present. The results of this study indicate that zinc supplementation during the immediate postinjury period is associated with improved rate of neurologic recovery and visceral protein concentrations for patients with severe closed head injury. PMID- 8714861 TI - Lesion volume, injury severity, and thalamic integrity following head injury. AB - Magnetic resonance scans of 63 TBI patients were analyzed to examine the relationship between injury severity, lesion volume (nonthalamic cortical/subcortical lesions), ventricle-to-brain ratio (VBR), and thalamic volume. For comparison, 33 normal control subjects were used. Patients with visible nonthalamic structural lesions showed significantly smaller thalamic volumes than patients without visible lesions or control subjects. Results also indicated that patients with visible lesions had significantly more severe injuries than patients without lesions. Patients with moderate-severe injuries had significantly smaller thalamic volumes and greater VBRs than patients with mild-moderate injuries. Although several variables related to thalamic volume, the presence of nonthalamic lesions was sufficient to result in smaller thalamic volume. Decreased thalamic volume following head injury suggests that subcortical brain structures may be susceptible to transneuronal degeneration following cortical lesions, and that this can be detected by in vivo MR-based volumetric analysis. PMID- 8714862 TI - Exposure to environmental complexity promotes recovery of cognitive function after traumatic brain injury. AB - This study was designed to determine whether exposure to a complex environment after traumatic brain injury (TBI) would promote the recovery of cognitive function. Rats were injured at a moderate level of fluid percussion injury (2.1 atm) or were prepared for injury but were not injured (sham injury). Immediately after the injury or sham injury, the injured/complex (n = 8) and the sham/complex (n = 7) groups were placed into a complex environment. The complex environment was a 89 x 89-cm enclosure with different types of bedding and objects that provided motor, olfactory, tactile, and visual stimulation. The injured/standard (n = 8) and the sham/standard (n = 8) groups were returned to the animal vivarium where they were housed individually in standard wire mesh cages (24 x 20 x 18 cm). On days 11-15 (postinjury), performance in the Morris water maze was assessed. Analysis of the latency to reach the goal platform indicated that injured animals recuperating in the complex environment performed significantly better than injured animals recovering in the standard environment (p < 0.01). In fact, injured animals in the complex environment performed as well as both sham injured groups. The improved performance of injured rats recovering in the enriched environment occurred in the absence of environmentally induced alterations in brain weight. These results indicate that exposure to environmental complexity enhances recovery of cognitive function after TBI. PMID- 8714863 TI - HU-211, a nonpsychotropic cannabinoid, produces short- and long-term neuroprotection after optic nerve axotomy. AB - HU-211 is a novel synthetic analog of tetrahydrocannabinol that was recently shown in animal models to be nonpsychotropic. In this study we show that HU-211 can potentially be used as a neuroprotective compound in the CNS. Using a calibrated crush injury of adult rat optic nerve, we show that HU-211 can reduce injury-induced metabolic and electrophysiological deficits. Energy metabolism was monitored by measuring the intramitochondrial nicotine-amine adenine dinucleotide redox state hourly for 6 h after injury and treatment. Electrophysiological activity was assessed by compound action potential and visual evoked potential response. Beneficial effects were dose-dependent, being optimal at 7 mg/kg, administered intraperitoneally. The time window during which treatment was effective was found to be from the time of injury for at least 5 h, with treatment most effective at the time of injury. These results strongly suggest that HU-211, given immediately after CNS injury at the optimal dosage, may possess neuroprotective activities. PMID- 8714864 TI - Perspectives on methylmercury as a global health hazard. PMID- 8714865 TI - The Seychelles study of fetal methylmercury exposure and child development: introduction. AB - Studies of outbreaks of methylmercury poisoning in Japan and Iraq from consumption of methylmercury (MeHg)-contaminated fish or bread proved that brain was the target organ, the toxic effects were dose-related, and the fetal brain was especially susceptible. Previous population studies suggested that a 5% risk of minimal fetal effect may be associated with a maternal hair mercury concentration during pregnancy of 10-20 ppm (mu g/g), a level that can be readily achieved by frequent consumers of fish. However, these studies had limitations, and as a result no definite conclusion concerning the lowest effect level could be reached and the question of a possible hazard to public health remained unanswered. There was a clear need for a more definitive study that would be prospective, involve a large enough cohort for risk analysis and adhere to accepted epidemiological principles. An appropriate site for such a study is the Republic of Seychelles, a location that afforded successful collaboration between the Seychelles Ministry of Health and faculty of the University of Rochester. A pilot study of 804 infant-mother pairs was followed by a main study of 779 mother infant pairs. In the pilot study children were examined once between 5 and 109 weeks of age; in the main study they are being evaluated longitudinally starting at 6 1/2 months of age. This paper introduces both the pilot and main studies, describes their design, and summarizes the findings through 6 1/2 months of age. When the Seychelles study is completed, the analyses will provide a database for those nations that choose to regulate their citizens' fish consumption and/or provide dietary education. PMID- 8714866 TI - The Seychelles child development study on neurodevelopmental outcomes in children following in utero exposure to methylmercury from a maternal fish diet: background and demographics. AB - Studies in Japan showed that fetal exposure to methylmercury during pregnancy can lead to severe neurodevelopmental changes in the infant while the mother suffers no or minimal effects. Fish contains methylmercury and there is concern that adverse neurodevelopmental effects may occur secondary to low-dose methylmercury exposure in utero from maternal fish consumption. The Seychelles Child Development Study has been examining the relationship between prenatal exposure to methylmercury during pregnancy in a population with high fish consumption and the neurodevelopmental outcome. Over 80% of Seychellois women eat fish daily, and the median fish meals per week during pregnancy is 12. Following a pilot study of 804 mother-infant pairs, a longitudinal main study of another 779 mother-infant pairs was initiated. The main study design includes collection of educational and socioeconomic information about the family and periodic standardized neurodevelopmental tests at specific ages from 6 1/2 months to 66 months of age. In this paper, we describe the background to the studies and give demographic characteristics of both the pilot and main study cohorts. PMID- 8714867 TI - The biological monitoring of mercury in the Seychelles study. AB - The concentration of total mercury in maternal hair during pregnancy was used as a measure of fetal exposure to methylmercury in a study of a fish-eating population in the Seychelles islands. A segment of scalp hair approximately 10 centimeters in length, that grew during pregnancy, was selected for measurement. Total and inorganic mercury were measured by cold vapor atomic absorption (CVAA) using the Magos reagents (Magos, 1971). For comparative purposes, total mercury was measured by X-ray Fluorescent Spectrometry (XRF) and methylmercury by gas chromatography/atomic fluorescence detection (GC/AFD) in a subset of hair samples. A limited number of fish samples were also analyzed. Extensive interlaboratory testing was conducted to ensure accuracy of the mercury measurements. Concentrations of organic mercury calculated as the difference between total and inorganic mercury as measured by CVAA agreed with those of methylmercury measured on the same samples of hair by GC/AFD. Methylmercury measured by GC/AFD and organic mercury measured by CVAA accounted for over 80% of the total mercury in hair and over 90% of the total mercury in fish muscle. To test the accuracy of recapitulation by hair sampling, hair samples were collected from mothers at the time of delivery and 6 months later. The segment corresponding to the pregnancy term was selected for measurement assuming a hair growth rate of 1.1 cm/month. Results from both samples were in close agreement. As part of both a pilot investigation followed by the main study, maternal hair samples were collected each year from 1986 to 1989 for a total of 1604 samples. The median and mean values for each year's collection fell in the range of 5.9 to 8.2 ppm and exhibited no statistically significant trend with time. The highest recorded concentration was 36 ppm. In hair samples from 654 mothers, the mean concentration of total mercury was compared with the mean concentrations in segments corresponding to each trimester, approximately 3.3 centimeters in length. A high degree of correlation was found between mean levels in each trimester versus the entire pregnancy segment. PMID- 8714868 TI - A pilot neurodevelopmental study of Seychellois children following in utero exposure to methylmercury from a maternal fish diet. AB - It is not known if fetal neurodevelopmental damage occurs in humans at the low level methylmercury exposure achieved by eating fish. To address this question, a cohort of 804 children in the Republic of Seychelles was identified who had fetal methylmercury exposure from a maternal diet high in oceanic fish. Mercury was determined by measuring the maternal total hair mercury during pregnancy, a standard index of methylmercury exposure. The median fetal mercury exposure was 6.6 ppm. Children were evaluated once between 5 to 109 weeks of age. Testing included the revised Denver Developmental Screening Test (DDST-R) and a neurological examination. The association between maternal hair mercury levels and developmental outcome was evaluated by multiple logistic regression analysis. Covariates for the child included gender, birth weight, one and five-minute Apgar score, age at testing, and medical problems, and, for the mother, age, tobacco and alcohol consumption during pregnancy, and medical problems. An association between fetal mercury exposure and development was found when DDST-R scores of questionable and abnormal were combined, a procedure used by previous investigators. These results should be viewed with caution since the association disappeared when DDST-R scores of questionable were treated in the standard manner as passes. PMID- 8714869 TI - Neurodevelopmental outcomes of Seychellois children sixty-six months after in utero exposure to methylmercury from a maternal fish diet: pilot study. AB - The Seychelles Child Development Study (SCDS) is testing the hypothesis that prenatal exposure to low concentrations of methylmercury from a maternal diet high in fish is related to the child's developmental outcome. In this report, 217 children from a pilot cohort were reevaluated at 66 months of age. The evaluation included the McCarthy Scales of Children's Abilities, the Preschool Language Scale, and age-appropriate sub-tests from the Woodcock-Johnson Tests of Achievement. Maternal hair total mercury, measured by cold vapor atomic absorption in a maternal hair segment corresponding to pregnancy, revealed a median exposure of 7.1 ppm. The association between maternal hair mercury levels and neurodevelopmental outcomes at 66 months of age was examined by multiple linear regression analysis with adjustment for important confounding variables. The results indicated that mercury exposure was negatively associated with four endpoints (the McCarthy General Cognitive Index and Perceptual Performance subscale and The Preschool Language Scale Total Language and Auditory Comprehension subscale). After normalizing the data by removal of a small number of outliers or highly influential scores, the mercury effects were no longer significant except for auditory comprehension. These results should be viewed as preliminary and interpreted with caution, since the SCDS main study 66-month evaluations, which are better controlled with more detailed endpoints are being analyzed. This study highlights the difficulties in interpreting epidemiologic studies of this type and the degree to which overall results in multivariate analyses can be influenced by a very small number of cases. PMID- 8714870 TI - Main neurodevelopmental study of Seychellois children following in utero exposure to methylmercury from a maternal fish diet: outcome at six months. AB - Methylmercury (MeHg) is a human neurotoxin to which the developing fetal brain is especially sensitive. The lowest dose of MeHg that impairs neurodevelopment in the human fetus is not known. The Seychelles Child Development Study (SCDS) is testing the hypothesis that fetal MeHg exposure from a maternal diet high in oceanic fish is related to child neurodevelopmental outcomes. Fish is the major protein source in the Republic of Seychelles, where a cohort of 779 mother-infant pairs was enrolled in a prospective longitudinal study. Maternal total hair mercury values during pregnancy were determined by cold vapor atomic absorption and ranged from 0.5 ppm to 26.7 ppm with a median of 5.9 ppm. When the children were 6 1/2 months of age, an examiner blinded to the maternal mercury value performed a neurological examination, the Fagan test of visual recognition memory, and the Denver Developmental Screening Test-Revised (DDST-R). On the DDST R 2% scored other than normal while 3.4% had an overall neurological score other than normal. The Fagan test of visual recognition memory showed a median score of 60.5%, and the Rose attention measure from that test showed a median score of 37.9. The association between fetal mercury exposure and neurodevelopmental endpoints was examined by multiple regression analyses. After adjusting for covariates, no association between the maternal hair mercury level during pregnancy and an adverse neurodevelopmental outcome of the child was identified at 6 1/2 months of age. PMID- 8714871 TI - Neurodevelopmental test selection, administration, and performance in the main Seychelles child development study. AB - The Seychelles Child Development Study was designed to provide data on normal neurodevelopment of Seychellois children and to examine the relationship of their neurodevelopmental outcomes to in utero fetal exposure to low concentrations of methylmercury from a maternal diet high in fish. This paper outlines the strategies used to select, modify, and field test evaluation tools used in the main longitudinal prospective study of 740 children (95% of the cohort of 779 initially enrolled in 1989). It also gives population statistics and quality assurance data for the tests administered and the evaluation of the home environments. PMID- 8714872 TI - Longitudinal neurodevelopmental study of Seychellois children following in utero exposure to methylmercury from maternal fish ingestion: outcomes at 19 and 29 months. AB - Despite the importance of defining developmental consequences for humans of in utero exposure to low levels of methylmercury, it is not yet clear if there are postnatal effects in fish-eating populations. The Seychelles Child Development Study (SCDS), now underway in the Republic of Seychelles, is following children to test the hypothesis that prenatal exposure to low concentrations of MeHg through maternal ingestion of fish is related to child development outcomes. In this study, children were evaluated with the Bayley Scales of Infant Development (BSID) at 19 mos. of age (N = 738). The cohort was evaluated again at 29 mos. (N = 736) with the BSID and the Bayley Infant Behavior Record. Mercury exposure determined by cold vapor atomic absorption analysis of maternal hair segments corresponding to pregnancy revealed a median exposure of 5.9 ppm (Range 0.5 - 26.7 ppm). The association between maternal hair mercury concentrations and neurodevelopmental outcomes at 19 and 29 mo. of age was examined by multiple regression analysis with adjustment for confounding variables. RESULTS: BSID Intertester reliability was ascertained by the Kappa statistic and was high. The mean BSID Mental Scale Indexes at both 19 and 29 mo. were comparable to the mean performance of US children. The mean BSID Psychomotor Scale Indexes at 19 and 29 mo. were 2 SD units above US norms, but consistent with previous findings of motoric precocity in children reared in African countries. No effect of mercury was detected on BSID scores at either age. On the Bayley Infant Behavior Record, activity level in boys, but not girls, decreased with increasing mercury exposure. Only one subjective endpoint was correlated with prenatal exposure to mercury. This study may have implications for environmental health policies concerning mercury in fish or fish consumption during pregnancy. Follow-up data are needed to determine if adverse effects occur at older ages and if such effects are determined to be related to mercury. PMID- 8714873 TI - An analysis of autopsy brain tissue from infants prenatally exposed to methymercury. AB - Brains from 32 neonatal autopsies from the Seychelles were examined histologically and analyzed for mercury levels. Six brain regions were sampled: frontal and occipital cortex, temporal cortex with hippocampus, basal ganglia with thalamus, cerebellum, and pons with medulla. Tissue blocks for histology and mercury analysis were taken from opposing faces to provide for correlation of findings. Similar studies were performed on 12 reference neonatal brains from Rochester, New York. No clear-cut developmental abnormality was found, but some brains exhibited low-grade, non-specific destructive changes. Total mercury levels, most of it in the organic form, were elevated in many of the Seychelles specimens. No correlation was demonstrated between mercury levels and degree or type of histologic change. There was considerable variability in total mercury for each anatomic region among the 32 Seychelles cases, as well as from one region to another in individual brains. All values of total mercury were under 300 ppb. Statistical analysis of mean mercury levels for each region demonstrated higher values in deep subcortical nuclei, brain stem, and cerebellum, phylogenetically older parts of the brain. When total mercury concentration of each region was paired with all other areas in the same brain and the paired values plotted for the entire group of brains, high correlations were obtained for all brain pairs, suggesting a strong concentration-dependent relationship between mercury intake and brain content. Analysis of mercury levels in separately dissected blocks of grey and white matter from 12 specimens revealed no significant differences between grey and white. In comparison with other human developmental studies and with experimental developmental studies in animals, where toxicity has been demonstrated with total mercury brain levels above 1,000 ppb, this study found no evidence of toxicity within a range of mercury levels below 300 ppb. Submicroscopic changes, subcellular alterations, subtle disturbances in the unfolding of brain architectonics -- none of these are excluded with methods used in this report. Further studies of threshold effects of MeHg on fetal brain are essential. That approximately half of the mercury resides in glial elements in white matter reinforces the need to focus attention upon glia as well as neurons during development. PMID- 8714875 TI - Summary of the Seychelles child development study on the relationship of fetal methylmercury exposure to neurodevelopment. AB - The Seychelles Child Development Study is examining the association between fetal methylmercury exposure from a maternal diet high in fish and subsequent child development. The study is double blind and uses maternal hair mercury as the index of fetal exposure. An initial cross-sectional pilot study of 804 infants aged 1 to 25 months suggested that mercury may affect development. A follow up of 217 pilot children at 66 months of age also suggested that neurodevelopmental effects might be present, but the result was dependent on outcomes in a small number of children. On the basis of initial results in the pilot study a prospective, longitudinal main study with more covariates and expanded endpoints was begun on a new cohort of 779 children. No association with neurodevelopment was seen at 6 1/2, 19, or 29 months of age, but there was an inverse relationship at 29 months in boys only between mercury level and activity as judged by the examiner. Adverse neurodevelopmental effects from fetal mercury exposure in the pilot study are highly dependent on how the data are analyzed and no definite effects have been detected through 29 months of age in the main study. In a related study, 32 brains were obtained at autopsy from Seychellois infants. These were examined histologically and analyzed for mercury. No clear histological abnormalities were found. Mercury levels ranged from a background of about 50 ppb up to 300 ppb, and correlated well between brain regions. For 27 brains maternal hair from delivery was available and hair mercury correlated well with brain mercury. PMID- 8714874 TI - Monitoring methylmercury during pregnancy: maternal hair predicts fetal brain exposure. AB - Autopsy brains were obtained from infants dying from a variety of causes within a few days of birth in a population exposed to methylmercury in fish. Infant and maternal blood and hair samples were also obtained. The concentration of total mercury in 6 major brain regions were highly correlated with maternal hair levels. This correlation was confirmed by a sequence of comparisons of maternal hair to maternal blood to infant blood and finally to infant brain. The results lend support to the use of maternal hair in assessing fetal exposure to methylmercury in fish-eating populations. PMID- 8714876 TI - Fetal methylmercury study in a Peruvian fish-eating population. AB - Maternal consumption during pregnancy of methylmercury (MeHg)-contaminated fish in Japan and of MeHg-contaminated bread in Iraq caused psychomotor retardation in the offspring. Studies in Iraq suggested adverse fetal effects when maternal hair mercury concentrations were as low as 20 ppm. This prospective study involved 131 infant-mother pairs in Mancora, Peru with peak maternal hair MeHg levels during pregnancy from 1.2 ppm to 30.0 ppm, geometric mean 8.3. The MeHg was believed to be derived from marine fish in the diet. There was no increase in the frequency of neurodevelopmental abnormalities in early childhood. The possible role of selenium or other protective mechanisms in marine fish is discussed. This previously unpublished study was conducted between 1981 and 1984. Our report of August 1985 to the funding agencies has been circulated, and the data were presented at the Twelfth International Neurotoxicology Conference in Hot Spring, Arkansas, October 30 to November 2, 1994. The current account has not been modified or updated since 1985. For reference to interim publications on fetal MeHg studies in Iraq and New Zealand see Marsh et al., 1995. PMID- 8714877 TI - Analysis of data on delayed development from the 1971-72 outbreak of methylmercury poisoning in Iraq: assessment of influential points. AB - In this paper we report additional results from the analysis of dose-response data on delayed development from the outbreak of methylmercury poisoning in rural Iraq during the winter of 1971-72. The discussion is focused on the importance of four influential points for the estimation of the population threshold for the effects of methylmercury. The point is made that while this study was important for establishing a dose-response relationship in humans, the uncertainty in the parameters of this relationship is large, and further human studies are needed. PMID- 8714878 TI - Collaboration between nurses and physicians: myth or reality? PMID- 8714879 TI - Telephone follow-up of gynecology day surgery patients. On line for quality and continuity of care. AB - Traditionally, patients who received minor gynecology surgery remained in the hospital for a few days, during which time post-operative care was given by nurses. However, since patients are now often discharged the same day, the responsibility for this care has been transferred to the patients and their families. Because the nursing staff of the Gynecology Day Surgery Unit at the Royal Victoria Hospital believes there is a need for patient-monitoring after discharge and CLSCs do not offer services for this generally healthy clientele, they developed, with the head nurse and the assistant head nurse, a post operative telephone follow-up program. This idea stemmed from the general need to adapt care as a result of the increase in day surgery. In addition to extending care beyond the hospital walls and improving its quality, this program has increased the job satisfaction of the nurses involved. PMID- 8714880 TI - [Clinical care for children and their families]. PMID- 8714881 TI - [Management of the length of stay of patients. Nurses presenting a memorandum to the administrative council]. PMID- 8714882 TI - [Nutrition a la carte]. PMID- 8714883 TI - [Susan Drouin, closely associated with the evolution of the nurse's role in neonatology. Interview by Helene Levesque]. PMID- 8714884 TI - [Discovering the nursing metaparadigm: care, person, health, environment]. PMID- 8714885 TI - [Allergy to natural latex. Strategies for preventive interventions and nursing care]. PMID- 8714887 TI - 1995 annual meeting of the National Council sets organization's course for coming year. Revisions to NCLEX-PN test plan adopted. PMID- 8714886 TI - [Parenteral hyperalimentation; a technique dependent on the nurse's competence]. PMID- 8714888 TI - An overview of CACCN Inc's history. PMID- 8714889 TI - Report from the twenty-second annual National Teaching Institute of the American Association of Critical-Care Nurses--New Orleans. (AACN NTI 1995 New Orleans). PMID- 8714890 TI - The development of the Australian Basic Intensive Care Knowledge Test. AB - The Basic knowledge Assessment Tool (BKAT), a test developed in the United States, has been presented as a valid and reliable test of basic knowledge for critical care nursing. However, it was necessary to determine the BKAT's validity and reliability in the Australian intensive care (IC) context. The Delphi technique, utilising a panel of eleven experts, was used to determine the content validity of the BKAT. The Delphi process resulted in the development of a test with 105 questions. These questions consist of 49 original BKAT questions, 25 original BKAT questions slightly modified, 3 original BKAT questions with major modifications and 28 new questions. A criterion group design was used to establish the modified test's reliability and decision validity. Item analysis was undertaken using item difficulty and item discrimination indexes. The modified test was completed by 14 registered nurses with no IC experience, 18 registered nurses with intermediate IC experience and 25 registered nurses qualified as intensive care specialists. The mean score (and standard deviation) for the test for each of the respective groups was 41(9), 69(9) and 86(7). These results were significantly different (p < 0001). The reliability was established with a Cronbach Alpha coefficient of .96. The modified test is a reliable and valid measure of IC basic knowledge and can be used as a valuable adjunct in the assessment of IC orientation programs. PMID- 8714891 TI - Quality improvement and critical care nursing. AB - Within the field of quality in health care there is a move away from quality assurance to a process of Continuous Quality Improvement. This change needs to be considered when designing a program to ensure quality within a critical care environment. There are several approaches available, one of which is the Dynamic Quality Improvement program. This involves solving problems and improving everyday practice whilst focusing on the recipients of the service. It utilises the Dynamic Standard Setting System (DySSSy) to define a professionally agreed level of performance for a chosen population. The standard must be achievable, observable, desirable and measurable. Patient focused nursing activities such as eye care and the prevention of pressure sores can be the basis for standard setting, audit and action to improve the care of the critically ill patient in a comprehensive program of quality initiatives. PMID- 8714892 TI - Computer-assisted learning (CAL) for general and specialist nursing education. AB - Computer-assisted learning (CAL) may be defined as any learning that is mediated by a computer and which requires no direct interaction between the user and a human instructor in order to run. Instead, CAL presents the user with an interface (constructed by an educator skilled in the field of study) which allows the user to follow a lesson plan or may allow self-directed access to particular information of interest. CAL has been claimed to improve knowledge retention and achievement scores, enhance clinical judgement skills and reduce required instruction time; performing as well (if not better) when compared to other more traditional education techniques. The advantages of the utilisation of CAL in nursing education can be made clear by consideration of adult education theory and curriculum design, as well as the particular learning needs of nurses themselves. Research and development into a theoretical framework for CAL design and implementation has allowed the identification of beneficial aspects of CAL resources. Although the cost of commercial software may be prohibitive to some institutions, possibilities exist for educators to create their own CAL packages relatively simply. PMID- 8714893 TI - Babies co-sleeping with parents. PMID- 8714894 TI - "Free speech". RCM stewards' network. PMID- 8714895 TI - Turn, baby, turn! PMID- 8714896 TI - Reach. The association for children with hand or arm deficiency. PMID- 8714897 TI - Nurses/midwives call for more HIV/AIDS protection. PMID- 8714899 TI - Staying the course in turbulent times. PMID- 8714898 TI - A letter from the '30s. PMID- 8714900 TI - [Learning nursing techniques in the clinical environment: a source of anxiety]. PMID- 8714901 TI - [Nursing care tied to the mourning process. Beyond palliative care]. PMID- 8714902 TI - [A program of nursing of the dying aged. A challenge in the field of voluntary services]. PMID- 8714903 TI - [Jeanne d'Arc Lebel. Inevitably life, love, death. Interview by Mariette Danis]. PMID- 8714904 TI - [The grief of nurses]. PMID- 8714906 TI - [From the power to tell to the power to do]. PMID- 8714905 TI - [Taking care of one's self]. PMID- 8714907 TI - [Health, social life and sexuality]. AB - This research study looked at the health, social life and sexuality of a portion of the University of Montreal student population. More than 600 resident and non resident students in the 18-26 age group were interviewed by the authors in an attempt to better define prevention strategies in the battle against AIDS and STDs. PMID- 8714908 TI - Self-directed learning experiences for senior BScN students. PMID- 8714909 TI - History lessons. AB - Interest in nursing history has grown in North America over the past two decades, in part because of the increase in feminist and nurse historians, but also because of the realization that nursing history can contribute much to the study of health and social issues. PMID- 8714910 TI - History of Canadian nursing: a legacy for the future. PMID- 8714911 TI - Enuresis treatment made simple. AB - Enuresis, the involuntary release of urine during sleep, is one of the most common disorders of childhood. Most children experience uncomplicated enuresis- the focus of this article. The condition, which is more common in boys than in girls, is characterized by nighttime wetting in the presence of a normal urinalysis and physical examination. PMID- 8714912 TI - Reducing the occupational risk of TB. AB - Tuberculosis, an infectious disease caused by the micro-organism Mycobacterium tuberculosis, has wrought devastation for centuries. According to estimates, in the 1800s alone, say estimates, it was responsible for almost one-third of the deaths in Europe. In the 20th century, the incidence of TB declined dramatically in developed countries, especially after the development of anti-TB drugs in the late 1940s. Yet in under-developed countries today, roughly one-third of the world's population is thought to be infected with the tubercle bacillus. PMID- 8714913 TI - Tuberculosis: the threat lingers. AB - Worldwide, tuberculosis has reached epidemic proportions. An estimated one-third of the world's population is infected with the disease. It kills three million people each year, and more adults die from TB than from any other infectious disease. In April 1993, The World Health Organization (WHO) declared a global TB emergency. For many people, this crisis state is incomprehensible. Didn't we get rid of TB years ago? PMID- 8714914 TI - [Family nursing interventions during hospitalization]. AB - More and more, families are being included in the nursing process. However, defining family nursing interventions still presents a challenge. Specific knowledge, abilities and attitudes are required to intervene with more than one person at a time. Observing the family in action enables the nurse to identify specific needs that generally fall into one of four types: cognitive, affective, physical and spiritual. These needs are then validated with family members to develop an appropriate care plan. If the care plan is based on Roy's model, family behaviors and contributing factors will be noted. If Orem's model is chosen, family abilities will be analysed to identify the problem solving difficulties family members have. A nursing diagnosis will then be made. As well, co-evolving objectives will take into account elements of change theory such as allowing time for change, being conscious of everyone's perception of the problem and not expecting every family member to change in the same way. Nurses play various roles with families. They offer support, and act as role models and educators. Their responsibilities can be shared with colleagues through care plans and nursing notes. PMID- 8714915 TI - [Learning to provide home care]. AB - More and more social policies are directed at community-based care. As a result, a family member, usually a woman, is called upon to become the caregiver of the sick person at home. In the case of a female caregiver who finds herself providing home care to a recently diagnosed cancer patient, major existential problems must be resolved. Without professional training, this woman must assume major responsibilities for the daily needs of her family member. To maintain a certain degree of control, she must learn to deal with her new situation. Facilitating the learning process becomes an important part of the nurse educator's role. By applying the Knowles theory of adult learning, the nurse educator is better able to understand the learning patterns of adults faced with such problems. This model helps develop a better perception of the adult learner, a clearer vision of the learning involved, and a better understanding of the nurse's role as educator. Before providing information, the nurse takes into account the perceptions of the caregiver and focuses on the person herself--thus recognizing her uniqueness. She helps the learner to become aware of her learning needs, to explore her motivations and to share her experiences. Through her attitude, the nurse communicates her intellectual and emotional availability, and encourages the caregiver to rely on and use her expertise. While providing necessary assistance, the nurse educator is instrumental in allowing the caregiver to take control of her own learning. PMID- 8714916 TI - A community approach to breastfeeding. PMID- 8714917 TI - Reduced resources and liability risks. AB - Cost-containment strategies have meant the loss of many registered nurse positions in health care facilities across Canada. Nurses are concerned about whether there will be enough adequately prepared professional health care staff to ensure safe, competent care consistent with professional nursing standards. Fears have arisen that substituting unregulated health care workers for nurses may pose a risk to the public that will lead to more lawsuits against nurses. PMID- 8714918 TI - Canada needs accessible masters' programs. AB - There are 14 masters in nursing programs in Canada, according to the Canadian Nurses Association. It sounds like enough, but unfortunately, most programs are inaccessible to nurses living outside the metropolitan regions. And though part time studies are offered at many sites, students must live near the universities. PMID- 8714919 TI - Managing health care. AB - By now we've heard about managed care at the client level, but what does the term mean in the context of the overall health system? As governments around the world strive to control health care costs, they are searching for better ways to manage the system and improve its efficiency. One approach has been the introduction of managed care models that integrate the financing and delivery of a range of health care services. So how do these models lead to better control and management of health care costs? PMID- 8714920 TI - The card. PMID- 8714921 TI - Research methodology. Part III: understanding multiple correlation analysis and multiple regression. AB - This is part III of a case series on research methodology with additional case demographic information. The relationship of a dependent variable with two or more independent variables is explained and used to illustrate multiple correlation and multiple regression. PMID- 8714923 TI - Videos and film clips as a medium for enhancing critical thinking. AB - Teaching as an art involves the use of varied methodology which is tried, true and creative. This article presents the use of video and film clips to develop/enhance critical thinking skills at the college level in two different disciplines, nursing and pre-service teacher education. PMID- 8714922 TI - The relationship between critical thinking and how time is structured in the clinical setting for students in a baccalaureate nursing program. AB - This experimental study was undertaken to investigate the relationship between critical thinking and how time is structured in the clinical setting. A total of 93 respondents completed the Watson-Glaser Critical Thinking Appraisal (CTA) Form: 35 in the comparison group, 29 in the 2-hour/8-hour clinical groups, and 29 in the 5-hour/5-hour clinical groups. Results of the ANOVA test shows that, at Time 1, students in the comparison group had the lowest mean score (9.23) and the 2-hour/8-hour clinical groups had the highest mean score (10.94), suggesting that the three groups were not equal at Time 1 in terms of their critical thinking ability. Nevertheless, the within subjects effects shows that while all three groups indicate positive Time 1-Time 2 changes, only the 5-hour/5-hour groups show a substantial degree of improvement (10.04 to 10.75), i.e., the study groups x time interaction effect is significant (p < .001). This pattern of significance between subjects effects and differential changes in CTA scores over time, with the 5-hour/5-hour groups showing the largest degree of change is consistent among the fine sub-component of the Watson-Glaser Scale. Students in the 5-hour/5-hour groups showed the most improvement in their scores in the area of Interpretation and Evaluation of arguments which was 9.55 at Time 1 and 10.52 at Time 2. PMID- 8714924 TI - Duplicate publication: guidelines for nurse authors and editors. PMID- 8714925 TI - The impact of the Occupational Safety & Health Administration regulations on management decision-making strategies. AB - Employee exposure to bloodborne pathogens is a potentially devastating event, for the institution and the employee. Managers, who have responsibilities to both the institution and the employee, can play a key role. Managers need to be thoroughly familiar with the Occupational Safety & Health Administration standard and their institution's exposure control plan. They must provide training to ensure compliance, initiate disciplinary measures for non-compliant employees, and identify potentially dangerous situations. In the event of an exposure, they must provide support to the employee. This article will describe the Occupational Safety & Health Administration standard and exposure control plan in detail, and will discuss the ways in which managers can ensure that the regulations are met and that employees are protected. PMID- 8714926 TI - Integrating the Occupational Safety & Health Administration mandates on bloodborne pathogens in the practice setting. AB - The Occupational Safety & Health Administration Bloodborne Pathogen Standard was developed to reduce the risk of exposure of healthcare workers to hepatitis B and C, human immunodeficiency virus, and other bloodborne pathogens. This can only be achieved if all healthcare workers continue to work to understand, adhere to, and enforce this mandate. The requirements of the Occupational Safety & Health Administration Standard will be discussed, including an exposure control plan, engineering and work practice controls, training, and hepatitis B vaccination. The risk to healthcare workers will be discussed in terms of the relevant pathogens, the extent of contamination, the risks associated with different occupations, and transmissibility. Finally, assessment and management of actual exposure incidents, as mandated by the Occupational Safety & Health Administration Standard, will be described. PMID- 8714927 TI - Intravenous modalities in the treatment of bloodborne pathogen illnesses: antibiotics, antifungals, and antivirals. AB - A variety of intravenous antimicrobial agents are used to treat infections with bloodborne pathogens. In using these agents to manage our patients, as well as any accidental employee exposure, it is important to understand how these agents work, and in which settings they are appropriate. This article will briefly review the historic development of antimicrobial agents and examine the agents according to the five basic mechanisms of action. Resistance to antimicrobial agents also will be discussed, as will the clinical and other factors that can positively or negatively affect the outcome of therapy. PMID- 8714928 TI - Patients, needles, and healthcare workers: understanding the epidemiology, pathophysiology, and transmission of the human immunodeficiency virus, hepatitis B and C, and cytomegalovirus. AB - The risk of transmission of bloodborne pathogens in the healthcare environment is determined by three main factors: the nature and frequency of exposure to blood or body fluids; the risk of transmission of infection after a single exposure to the pathogen; and the prevalence of infected and susceptible patients and healthcare workers. This article will examine all three parts of the risk equation by addressing the risks and pathologic consequences of infection with four important bloodborne viruses: human immunodeficiency virus, hepatitis b, hepatitis C, and cytomegalovirus. Approaches to the management of occupational exposures to bloodborne pathogens and prospects for prevention also will be discussed. PMID- 8714929 TI - The challenges of teaching universal precautions to multicultural, diverse patients and their family members. AB - Teaching universal precautions to culturally diverse groups, whether healthcare workers, patients, or family members, is a unique challenge. In the environment of acquired immune deficiency syndrome, it involves honest and realistic appraisal of the modes of transmission and the risks involved. In addition, we must understand the fears involved, as well as the cultural perspectives that may alter the perception of the situation. This article will discuss the many cultural problems, briefly review what information needs to be included in any educational program, and focus on how we can realistically assess the risks involved and communicate those risks to healthcare workers, patients, and family members. When these risks are addressed honestly and without fear, implementation of and compliance with universal precautions will become easier, both in acute care and home care settings. PMID- 8714930 TI - Ethical issues in the handling of bloodborne pathogens: evaluating the Occupational Safety & Health Administration Bloodborne Pathogen Standard. AB - The Occupational Safety & Health Administration Bloodborne Pathogen Standard is designed to help provide protection from contamination in the workplace. Healthcare professionals must evaluate the standard and the behaviors mandated therein, according to the basic professional ethical principles of professional autonomy, beneficence, and cost-benefit considerations. The patient's wellbeing provides the focus for making such an assessment. PMID- 8714931 TI - The development of a risk management program in response to the spread of bloodborne pathogen illnesses. AB - Risk management and total quality management are both focused on prevention, and together they provide the means to achieving improved outcomes. In total quality management, we are constantly looking for ways to improve processes, by continuously evaluating systems, identifying root causes, and providing employees with training, resources, and empowerment. Total quality management identifies opportunities for improvement, and risk management is used to prioritize these. Risk management involves identification and analysis of risk, development of action plans to deal with risk, examination of financial issues, and constant evaluation of the system. This article will describe the development and implementation of a risk management program, both in general and regarding bloodborne pathogen illnesses. PMID- 8714932 TI - Student's perceptions of psychiatric nurses' roles. PMID- 8714933 TI - Raising the profile of Maori. PMID- 8714935 TI - The increasing regulation of nursing. PMID- 8714936 TI - Being a good employer: what does it mean? PMID- 8714934 TI - Reforms round-up. Where the money goes. PMID- 8714937 TI - Public beds for private patients. PMID- 8714938 TI - Storytelling at work. PMID- 8714939 TI - Primary nursing does have its place. PMID- 8714941 TI - Deafness offers new insights to care. Interview by Sue Townsend. PMID- 8714940 TI - A nurse's story. PMID- 8714942 TI - The best of times and the worst of times... PMID- 8714943 TI - Who leads nursing in 1995? PMID- 8714945 TI - Nursing care of infants on phototherapy. PMID- 8714944 TI - The stories must be told. PMID- 8714946 TI - Tips on resume writing. PMID- 8714947 TI - Graduate education in nursing. PMID- 8714949 TI - Questions and answers on preparing for the NCLEX. PMID- 8714948 TI - Nursing assistant jobs; a strategy for experience and employment. PMID- 8714950 TI - Why I chose nursing as a career. PMID- 8714951 TI - Community health nursing internships for new generic graduates: the University of Kentucky. PMID- 8714966 TI - Update on NCLEX using computerized adaptive testing (CAT). PMID- 8714968 TI - Delegation: concepts and decision-making process. National Council of State Boards of Nursing, Inc. PMID- 8714967 TI - An NCLEX item reviewer shares her experience. PMID- 8714969 TI - Thoughts on nursing roles and keeping the workforce intact. PMID- 8714970 TI - Every patient deserves a nurse. PMID- 8714971 TI - Tis the season to be ... codependent! And stressed! Or how to get over yourself. PMID- 8714972 TI - Nurses can be heard--here is how: grassroots lobbying tips. PMID- 8714973 TI - Children and health. PMID- 8714975 TI - Advanced practice nurse practitioners: a legacy of leadership. PMID- 8714974 TI - Look alike/sound alike drug names. PMID- 8714976 TI - Background checks for nurse aides. PMID- 8714977 TI - Dear Nurse Newheart. PMID- 8714978 TI - Changes in the program for impaired nurses. PMID- 8714979 TI - Peer assistance. PMID- 8714981 TI - Can you compete? PMID- 8714982 TI - Registered nurse perceptions of nursing practice. PMID- 8714980 TI - How to communicate effectively with your legislator. PMID- 8714983 TI - First national EN conference a success. PMID- 8714984 TI - ANF national delegates conference. PMID- 8714985 TI - Saving money with waste. PMID- 8714986 TI - The cultural life of a hospital. PMID- 8714987 TI - Are level 2 clinical nurses being used appropriately? PMID- 8714988 TI - Back to basic values. PMID- 8714989 TI - RNABC's third annual member survey. PMID- 8714990 TI - Nurses as partners in care. Going the distance for women with breast cancer. PMID- 8714991 TI - Nurses as partners in care. Teaching women self-defence. PMID- 8714992 TI - Nursing centre; practicing what we preach. PMID- 8714993 TI - Standards in action: maintaining competence. PMID- 8714994 TI - Using research to improve the quality of nursing care. PMID- 8714995 TI - A chapter with a view. PMID- 8714997 TI - Nursing in the year 2000. PMID- 8714996 TI - Dispensing revisited. PMID- 8714998 TI - October is Breast Awareness month. New Mexico Breast and Cervical Cancer Detection and Control Program. PMID- 8714999 TI - Meeting with AHCA director of medical quality assurance addresses nursing issues. PMID- 8715000 TI - Ideas on leadership. PMID- 8715001 TI - Journey of discovery. PMID- 8715002 TI - Through the eyes of a student. PMID- 8715003 TI - Inclusion of students with disabilities in nursing education programs in Florida. PMID- 8715004 TI - Professional nursing and unlicensed assistive personnel: a practice dilemma for the 90s. PMID- 8715005 TI - Staff involvement critical in enhancing a safe environment for care. PMID- 8715006 TI - Rx: mentorship--the drug of choice for professional FTT. PMID- 8715007 TI - News along the Rialto. PMID- 8715008 TI - "Caring" is not enough: ethical paradigms for community-based care. PMID- 8715009 TI - Ethics, nursing, and health care in the age of "re-form". PMID- 8715010 TI - Clinical failure or clinical folly? A second opinion on student performance. PMID- 8715011 TI - Research & nursing: ethical reflections. PMID- 8715012 TI - Art in practice. The Attentive Nurse. PMID- 8715013 TI - President's message: riding the breaker waves. PMID- 8715015 TI - Dressings for work. PMID- 8715014 TI - Post-raid debriefing. PMID- 8715016 TI - Seeing the light. PMID- 8715017 TI - The best laid schemes. PMID- 8715018 TI - Discrimination is bad news. PMID- 8715019 TI - Philanthropic Societies. PMID- 8715020 TI - Aqueous shunt implant surgery for refractory glaucoma. AB - The newer generation of aqueous shunt implant for glaucoma filtration surgery has increased our ability to manage more successfully glaucoma refractory or uncontrollable by medical therapy or standard surgical means. These clinical results suggest that surgery with such devices may even provide greater efficacy than standard filtration operations. The development of modern sophisticated device designs incorporating unidirectional valves has reduced complication and re-operation rates. Confidence in the use of these newer devices will increase as surgeons gain wider experience. Presently, some surgeons are considering aqueous shunt implant procedures before resorting to adjunctive antimetabolites to avoid scleral degeneration that may limit subsequent shunt surgery. Although glaucoma remains a disorder without a cure, the advent of aqueous shunt implants for glaucoma filtration surgery has greatly increased the prognosis for this visually threatening disorder. PMID- 8715021 TI - Glaucoma: an overview. AB - Glaucoma is one of the leading causes of blindness. It is a public health problem. No race is exempted from getting glaucoma. In the United States there are more than 2 million people afflicted with this disease. Every effort should be made to detect those people at risk early, and adequately manage them to prevent blindness. Therefore, it is important than an awareness of this disease should be created among health care professionals and the general public. This will lead to the early detection and proper management of this disease to prevent blindness. Glaucoma is a group of ocular conditions that will lead to blindness if it is not controlled. Although our knowledge about this group of diseases has advanced much in the past few decades--as compared with earlier times--we still are ignorant about the finer points and details regarding the disease. A lot of basic and clinical research is required to further and completely understand it. Much research is being done toward better management of glaucoma. Many multicenter clinical trials are being conducted. Collection of data regarding glaucoma is ongoing and will contribute to a better understanding of glaucoma- and thereby better management of patients who develop glaucoma. PMID- 8715022 TI - Freezing of preservation media during corneal tissue transport. AB - It is important to evaluate donor corneas and media carefully prior to surgery. The eye banks that are responsible for preservation and transportation of donor corneal tissue should continue their efforts to prevent tissue damage, which includes careful placement of vials in properly prepared containers. If placed in direct contact with ice, the ice should be relatively warm and glistening. Direct contact with hard, super-cooled ice should be avoided. If living tissue is frozen without special treatment, ice crystals form in the cells and cell membrane rupture, causing death of the cell. Also, when ice crystals are present, tissue damage could take place when physical impact is caused by motion of the container during transportation. PMID- 8715023 TI - Silicone oil's properties make it a versatile tool for vit/ret surgery. AB - Silicone oil initially was perceived as a surgical tool to separate proliferative tissue from the retinal surface. The role of silicone oil as an intraocular tamponade became more clearly established following the introduction of trans pars plana vitrectomy in 1970 and the significant subsequent refinements in surgical techniques and technology. Silicone oil maintains an important place in the vitreoretinal surgical armamentarium. It may help preserve sight in some otherwise inoperable eyes. Future research may utilize silicone oil as a reservoir for the slow release of a therapeutic concentration of lipophilic drugs that inhibit viral retinitis or the proliferation of epiretinal membranes. PMID- 8715024 TI - Computers and eyestrain. AB - Computer video display terminals have revolutionized the home and office work habits of millions of people. Although no verifiable organic ocular diseases have been shown to derive from computer monitors, symptoms related to eyestrain are very common. Some 10% to 15% of patients presenting for routine eye exams complain of computer-related headache and eyestrain. Comprehensive ocular examination including distance, intermediate and near refraction, as well as uncovering evidence of ergonomic and lighting inadequacy, often is clinically quite helpful. A careful eye exam with particular attention to middle and near range refraction, oculomotor balance, the workstation, lighting, and seating comfort will assist measurably in alleviating eyestrain. PMID- 8715025 TI - An interview with Margaret Koffler, RN, BSN, MAEd. Interview by Todd A. Hostetter. PMID- 8715026 TI - A new look at extended wear. PMID- 8715027 TI - What's your diagnosis? Astrocytic hamartoma. PMID- 8715028 TI - [Decreasing the risk]. PMID- 8715029 TI - [STANDARD enteral feeding using a catheter or feeding pump in patients with a gastric tube]. PMID- 8715030 TI - [Pathogenesis and differential diagnosis of "diabetic foot lesions"]. PMID- 8715031 TI - [How well does diabetes have to be controlled?]. PMID- 8715032 TI - [Resuscitation by hospital personnel. Self assessment and educational status]. PMID- 8715033 TI - [High fluid absorption with alginates. Better quality with less work]. PMID- 8715035 TI - [Impulses from Switzerland--electronic service planning]. PMID- 8715034 TI - [Thrombolysis therapy in cerebral ischemia]. PMID- 8715037 TI - Scope and standards for nurse administrators. American Nurses Association. PMID- 8715038 TI - Statement on the scope and standards of pediatric clinical nursing practice. American Nurses Association. PMID- 8715039 TI - On getting consent: how do recent legislative changes on consent affect you? PMID- 8715040 TI - Organizational restructuring. PMID- 8715041 TI - Computer connection. PMID- 8715042 TI - Nurse-client relations: how far is too far? PMID- 8715043 TI - Promoting good practice. PMID- 8715044 TI - Preventing undesirable practice. PMID- 8715045 TI - Intervening when necessary. PMID- 8715046 TI - Rural/frontier nursing: the challenge to grow. PMID- 8715047 TI - Importance of human relations in providing effective nursing care. PMID- 8715049 TI - Community health nursing: an overview of teaching and training. PMID- 8715050 TI - The genome imperative. PMID- 8715048 TI - Sex education for adolescents. PMID- 8715051 TI - Genetic information and health insurance: state legislative approaches. PMID- 8715052 TI - Genetic privacy. PMID- 8715053 TI - Testing children for genetic predispositions: is it in their best interest? PMID- 8715054 TI - Beyond "genetic discrimination": toward the broader harm of geneticism. PMID- 8715055 TI - Conflicts in the biotechnology industry. PMID- 8715056 TI - Drafting the Genetic Privacy Act: science, policy, and practical considerations. PMID- 8715057 TI - The attempt to pass the Genetic Privacy Act in Maryland. PMID- 8715058 TI - Legislating privacy: the HIV experience. PMID- 8715059 TI - Why the use of anonymous samples for research matters. PMID- 8715060 TI - The impact of the Genetic Privacy Act on medicine. PMID- 8715061 TI - Ethical issues raised by needle exchange programs. PMID- 8715063 TI - Ethics consultation: anencephaly and organ donation. PMID- 8715062 TI - Stemming the tide: assisted suicide and the constitution. PMID- 8715064 TI - Nurses and consumers: partners in assuring quality care in the home. PMID- 8715065 TI - Nursing tasks today. PMID- 8715066 TI - Doing transcultural nursing research. PMID- 8715067 TI - [AIDS: rights and obligations, equally divided]. PMID- 8715068 TI - [Organizing preventive measures]. PMID- 8715069 TI - [School vaccination. Experiences in the local dispensary]. PMID- 8715070 TI - [The nurse and the children (II). Identity, criticism and ethics in children's toys and books]. PMID- 8715071 TI - [Education in geriatric dentistry]. PMID- 8715072 TI - [How to make them listen to what they do not want to hear: use of cultural tradition as a methodological instrument for learning the history of nursing]. PMID- 8715073 TI - [Controversy on the effectiveness of vaccinations]. PMID- 8715074 TI - [The interchange of residents: a felicitous initiative]. PMID- 8715075 TI - [INTERNET (Y II). Aids to nursing]. PMID- 8715076 TI - [Cutaneous distribution of the thoracic nerves]. PMID- 8715077 TI - [Health problems cared for by nurses of the primary health care teams]. PMID- 8715078 TI - [Natural family planning. An alternative]. PMID- 8715079 TI - [Prostatic changes. Preventing complications]. PMID- 8715080 TI - [When we need imagination]. PMID- 8715082 TI - [Standardized control of vital signs in a surgical intensive care unit]. PMID- 8715081 TI - [Symptoms depending on the menstrual cycle in female personnel at an university hospital]. PMID- 8715083 TI - [Planning and organization of a newly formed ward]. PMID- 8715084 TI - [Comparative study after conventional and laparoscopic removal of the gallbladder]. PMID- 8715085 TI - [Basic concepts in the treatment of patients with chronic pain]. PMID- 8715086 TI - [Adaptation to change]. PMID- 8715087 TI - A call on geriatric nursing. PMID- 8715088 TI - Leadership development and training in nursing and midwifery. PMID- 8715089 TI - Nursing education for the betterment of family health. PMID- 8715090 TI - Integrated health care. A program to enhance nurse-physician relations. PMID- 8715091 TI - Managing in an age of discontinuous change. PMID- 8715092 TI - Finding the balance. PMID- 8715093 TI - Case management: the promise, the process, and the vision. PMID- 8715094 TI - [Cystic fibrosis. Epidemiology, physiopathology and symptoms]. PMID- 8715095 TI - [Cystic fibrosis: current treatment and perspectives]. PMID- 8715097 TI - [Cystic fibrosis: enteral nutrition at home. Role of the coordinating nurse]. PMID- 8715096 TI - [Cystic fibrosis: nutritional strategy]. PMID- 8715098 TI - [The pump for refrigerated nutrition]. PMID- 8715099 TI - [Cystic fibrosis. The nurse coordinating the care plans]. PMID- 8715100 TI - [Cystic fibrosis: the nurse coordinating home care. Intravenous antibiotic therapy at home]. PMID- 8715101 TI - [Gene therapy; a necessity]. PMID- 8715102 TI - [Encopresis]. PMID- 8715103 TI - [Children and death]. PMID- 8715104 TI - [Test tube babies and illness]. PMID- 8715105 TI - [Pediatric emergencies: heart failure]. PMID- 8715106 TI - Teaching RNs how to select a nursing assistant partners. AB - In today's world of health care restructuring, most RNs will work with a Nursing Assistant (NA) or other unlicensed nursing personnel. Problems from lack of the RN's supervision skills can create difficulties in the NA's performance, interfere with the working relationship of the RN/NA pair, and decrease nurse satisfaction in working with assistants. Teaching staff to choose a NA partner is an important first step in increasing the RN's ability to supervise the NA. This experienced author describes key strategies for teaching RNs interviewing and hiring skills needed for skillmix changes and partnerships. PMID- 8715107 TI - Dispelling myths about the new NCLEX exam. AB - The new computerized NCLEX system is working well. Most new candidates, employers, and board of nursing representatives like the computerized adaptive testing system and the fast report of results. But, among the candidates themselves some myths have grown which cause them needless anxiety. PMID- 8715108 TI - [Caring for myocardial infarction of simple evolution]. PMID- 8715109 TI - [Care of myocardial infarction with complications]. PMID- 8715110 TI - [Coronary angiography and coronary dilatation]. PMID- 8715111 TI - [Care of a patient treated by fibrinolysis for myocardial infarct]. PMID- 8715112 TI - [Hospitalization for myocardial infarction]. PMID- 8715113 TI - [Intensive care in cardiology]. PMID- 8715114 TI - [Mobilizing patients. Spirit and practice]. PMID- 8715115 TI - [Neurotic disorders in old age. A psychodynamic approach]. PMID- 8715116 TI - [Diseases transmissible by blood. Occupational risks and prevention]. PMID- 8715117 TI - [Anthropology of pain. Notes after reading the work of David Le Breton]. PMID- 8715118 TI - [The clinical aspect of nursing care]. PMID- 8715119 TI - [The LH-RH analogs. Drug treatment of prostate cancer]. PMID- 8715120 TI - Charge nurses: critical change agents for successful restructuring. AB - Charge nurses know a unit's needs, difficulties, strong points, and successes. Therefore, charge nurses are often the key people that make restructuring work at the unit level. This article describes how to empower charge nurses so they can be the drivers for today's changes. PMID- 8715121 TI - Future nursing career fair: helping nurses prepare for the future. AB - Nursing administrators today are faced with the problems of downsizing, loss of current nursing jobs, and discouraged nurses. A program to show growth areas in health care can help nurses look toward the future to expand and build expertise in the growing fields. This article describes steps in developing a medical center based, career fair program for nurses. PMID- 8715123 TI - Nurses take lead in consumer education. PMID- 8715122 TI - Report from nursing practice--what Alberta nurses are asking about. PMID- 8715124 TI - ANA challenges Pew Health Professions' findings. PMID- 8715125 TI - 1996--speak out for quality care. PMID- 8715127 TI - Unity serves as key to promote nursing's agenda. PMID- 8715126 TI - Who's taking care of Mama? As I see it. PMID- 8715128 TI - OSHA targeted for "reform". PMID- 8715130 TI - Poll shows SNA members more liberal than public. PMID- 8715131 TI - ANA works to prevent medication errors. PMID- 8715129 TI - MSNA conducts nursing research survey. PMID- 8715132 TI - Feds threaten to nix NLN's accreditation. PMID- 8715133 TI - Conference stresses crucial role of vaccines. PMID- 8715134 TI - CDC funds immunization grant: a nursing implementation for vaccine service. PMID- 8715135 TI - ANF project addresses HIV, mental health link. PMID- 8715136 TI - Euthanasia: the slippery slope. PMID- 8715137 TI - So much is at stake. PMID- 8715138 TI - Casemix--only part of the big picture. PMID- 8715139 TI - Nurse leaders heading for parliament. Interview by Amanda Tattam and Sam Prenesti. PMID- 8715140 TI - Indonesia: workers fight to join a union. PMID- 8715142 TI - A new path to documentation. PMID- 8715141 TI - Nursing and the law. Dealing with sexual harassment. PMID- 8715143 TI - [Nurses in hospital hygiene...]. PMID- 8715144 TI - [The fight against nosocomial infections. French policies]. PMID- 8715145 TI - [Nosocomial infections. Organization of care within a policy of prevention]. PMID- 8715146 TI - [Nosocomial infections. Introduction of a program of supervision]. PMID- 8715147 TI - [Prevention of nosocomial infections. Place of a manager in a center of coordination]. PMID- 8715148 TI - [Implantation site and risk of infection]. PMID- 8715149 TI - [Short venous catheters. Evaluation of the quality of its setting and supervision]. PMID- 8715150 TI - [Cleaning and disinfection of endoscopic material. 1. Current recommendations]. PMID- 8715151 TI - [Cleaning and disinfection of endoscopic material 2. How to unify techniques for flexible and rigid endoscopes in a CHU?]. PMID- 8715152 TI - [Why do we not have an optimal model of hospital hygiene! Experience of an interaction between 2nd year nursing students and the nursing service]. PMID- 8715153 TI - [How to make one's home safe against theft]. PMID- 8715154 TI - [Young and dynamic nurses]. PMID- 8715155 TI - [Among coconuts and bananas]. PMID- 8715156 TI - [Reflections about the changes in our society (1). A civilization to be invented]. PMID- 8715157 TI - [Caring for depressed patients. Should we leave them alone or activate them?]. PMID- 8715158 TI - [Colloquium "life in an institution, institutional life. Encouraging living projects]. PMID- 8715159 TI - [Good education is no protection from unemployment. The uneducated unemployed--a prejudice]. PMID- 8715160 TI - [What kind of a world do we want?]. PMID- 8715161 TI - [Parents at the pediatric intensive care unit. With guidance and sympathy]. PMID- 8715162 TI - [Visit of clowns in a pediatric hospital. Laughing and cheerfulness in spite of illness]. PMID- 8715163 TI - [How did the clowns get into the hospital?]. PMID- 8715164 TI - [In the obstetrics and gynecology department of the Eritrea hospital "Place for Life", even the twelfth child is precious]. PMID- 8715165 TI - [Introduction of new hygiene guidelines. The resistance to change]. PMID- 8715166 TI - [Motivation of aged residents of homes ... so they will again participate in life]. PMID- 8715167 TI - [Language must not degenerate into unpronounceable bilge]. PMID- 8715168 TI - [A difficult task successfully solved]. PMID- 8715169 TI - [A patient from a different culture. Care in the intercultural reality]. PMID- 8715170 TI - Family caregivers: a crucial element in our long-term care system. PMID- 8715171 TI - Struggling to find the exit in the dark. AB - Every one must learn to deal with a devastating illness such as Alzheimer's disease in his or her own way. When so little is known, however, each person's experiences can add to the knowledge of us all. In this article, the author eloquently urges us to find ways to maintain the humanity of the dementia impaired--to see beyond bizarre behavior or very physical needs and remember the mind and soul that remain there still. The author offers professionals and nonprofessionals alike a poignant, yet positive, view of what is possible in a seemingly impossible situation. PMID- 8715172 TI - To care, not to cure. AB - With a diagnosis "presumed" to be Alzheimer a commitment was made by the family to focus on care; to help Virginia deal with daily living and to help her find as much enjoyment for the rest of her life as possible. As she moved toward increasing dependence over the 16 plus years, her independence and other qualities were protected and nurtured. Virginia was kept stimulated by being integrated into the daily schedule of the primary caregiver while at the same time supporting her special needs. This paper is not intended as a statement against institutionalization but suggests that caregiving can be managed and integrated into the normal routine of life. PMID- 8715173 TI - A case manager's practical tips for family caregivers. AB - Care plans are developed to meet the needs of persons suffering from a dementia or a chronic, debilitating disease. Nurses, social workers, physicians, physical therapists, and other health care professionals create the care plans, primarily with the care recipient in mind. Yet, often, it is the family caregiver who must provide the majority of home care. Who considers this caregiver's needs? This article focuses on the informal, family caregiver, often untrained, who must follow the care plan, with or without help from the professionals. It offers practical suggestions to help these caregivers carry out their role with productive and satisfying results. PMID- 8715174 TI - The dilemmas of dependency. AB - Dependency can engender a host of negative feelings. Anger or irritation at caregivers, a burdening sense of indebtedness, and fear of increasing frailty are emotions commonly experienced by those receiving care. These negative reactions may stem from early life experiences in which parents failed to fulfill basic needs for protection and nurturance. Older patients who were abusive to their children when young may worry about "paybacks" during dependency. Still others recall the strain of taking care of their own parents years earlier and are afraid of anyone feeling toward them what they once felt. Differing expectations between generations, as well as sibling conflicts over parental care, can add to the difficulties. Case managers who are aware of these potential issues can help clients through them to a resolution. PMID- 8715175 TI - Gerontology for companions: a model curriculum. AB - Although paid companions spend many hours in direct contact with their older clients, the majority have little or no grounding in gerontological developmental theory. This article describes a project that provided academic training for paraprofessionals through a pilot curriculum. Goals were to provide companions who were already working with socially isolated clients with a sound theoretical and practical grounding in communication skills, the aging process, and issues of loss and grief. To accomplish these goals, partnership was developed between an academic institution, a case management agency, and a direct service provider. Preliminary analysis of the pretest and posttest data measuring the outcome of the modules found that, as companions learned about continued growth and development in their older clients, these workers achieved a greater sense of self-fulfillment and job satisfaction. PMID- 8715176 TI - Service use by caregivers of elderly receiving case management. AB - This article reports on findings from survey interviews with 401 informal caregivers of impaired older persons who were clients of nonprofit case management agencies in Cleveland, OH. The issue addressed is the role of the case manager as a facilitator of caregiver service use when the older person receives case management service. Caregiver services under investigation include respite care, help finding or arranging for service, emotional support or counseling, caregiver education and training, and assistance with other responsibilities such as child care. The study questions are: to what extent do caregivers in this sample utilize these services? What are the predictors of caregiver service use? What implications do the findings have for case management practice? PMID- 8715177 TI - When "the family" is not one voice: conflict in caregiving families. AB - Caregiving families rarely have one voice. This article presents a theoretical framework for normative family conflict in chronic care, identifies types of family conflict, and describes educational and case management models to prevent, identify, buffer, or potentially help resolve family care conflicts that pose barriers to care. Successful application of these models can result in greater family member accessibility for support to each other and enhanced family capacity to distribute equitably and effectively care responsibilities for chronically ill relatives especially those with Alzheimer's disease and other dementia. PMID- 8715178 TI - Family caregiver education and training: enhancing self-efficacy. AB - Families who face the necessity of caring for an elderly family member need information and education. In particular, case managers and other professionals can provide information in seven key areas: the care receiver's medical condition; coping skills; family disruption, anger, and other relationship issues; communication; community services; emotions, including guilt; and long term planning. Each family will have its priorities, and professionals assisting the caregiver need to avoid assuming one issue is more important than another without consultation. Education through videocassettes, audiocassettes, printed materials, workshops, and one-on-one sessions can be a powerful influence on the caregiver's self-efficacy and confidence. Stress reduction, diminished guilt feelings, and more satisfactory family relationships are among the potential results. PMID- 8715179 TI - Facilitators and barriers to caregiver support group participation. AB - This study examined the facilitators and barriers to support group participation among lower socioeconomic status African-American and Caucasian caregivers of persons with chronic mental illness. Data were collected through in-person interviews with family caregivers of 198 adults with chronic mental illness. The results indicated that a substantial numbers of caregivers had little knowledge about support groups. Mental health professionals were cited as important facilitators of support group participation. Barriers to support group participation pertained to accessibility and personal factors including lack of time, lack of need, difficulties sharing feelings, concern for confidentiality, and not knowing anyone who was a group member. There were few differences by caregiver race. Implications for mental health case managers and agencies and for future research are discussed. PMID- 8715180 TI - [Thalassemia major--home care for a 6 year old boy]. PMID- 8715181 TI - [Baptism--emergency baptism]. PMID- 8715182 TI - [Cooperation--organization--communication]. PMID- 8715183 TI - [Care of dying children--it raises our anxieties facing finiteness and stresses our feelings]. PMID- 8715184 TI - [Thermal care in the thermal bed with a gel-mattress]. PMID- 8715185 TI - [The problem of HIV infections in children in the daily life in the hospital]. PMID- 8715186 TI - [Confidentiality--the basis of trust in nursing. I]. PMID- 8715188 TI - [Main points in hospital care. Discussions by the governmental coalition on the third phase of Health Reform]. PMID- 8715187 TI - [Toxic and less toxic plants. 23]. PMID- 8715189 TI - [Preparation for intramuscular injections]. PMID- 8715190 TI - [Hepatitis B as an occupational risk]. PMID- 8715191 TI - [The use of nursing records in the Veneto region]. AB - Use and quality of nursing records used in the Regione Veneto hospitals was evaluated. An open ended questionnaire was sent to the nurses managers of each hospital. The questionnaire asked some general information on the hospital (number of beds, number and qualification of nursing personnel) and nurses managers were asked to send a blank copy of the nursing records in use. 21 nurses managers answered, overall responsible for 28 hospitals. Nurses records are used in 78 wards over 412 (19.1%). The wards are different for specialty and characteristics. Apparently, the only factor associated with the implementation of nursing records is the number of beds, with a wider implementation of nursing records in smaller hospitals. No association was found with the level of qualification of the nursing personnel. 11 nursing records were received. The items to be compulsorily monitored were mostly physical needs (i.e. elimination, hygiene, nutrition, mobilization), with the exception of the two records used in the psychiatric wards, where data on the behaviour and reaction of the patient was available. PMID- 8715193 TI - [Cases: when the hospital becomes alive]. PMID- 8715192 TI - [Epidemiology in nursing routine: data and hypotheses of longterm elderly patients]. AB - Drugs prescription in patients > or = 65, admitted to the Geriatric ward (48 beds) of the Hospital of Crespano del Grappa (VC) from 1st january to 15th October 1993, was retrospectively analyzed examining clinical records. Respiratory and cardiovascular problems affect 18% of patients. The most frequently prescribed at discharge are cardiovascular drugs (71%) and gastrointestinal drugs (23% in women and 18% in men). Only 10 active principles (over 150 prescribed) lack a scientific documentation of efficacy. The survey is presented as one of the ways for producing an epidemiological profile of the patients cared for, beginning from easily available data. Such a profile would allow to focus on specific problems (the different pharmacologic management of men and women; the complex patients; the patients that would need specific information on drugs), and plan intervention strategies. PMID- 8715194 TI - [Methods and instruments of research: analysis of various articles]. AB - Research papers published on some of the most well known medical sud nursing journals are presented and discussed. The main aim of the contribution, which opens an arena for discussion on the Rivista dell'Infermiere is to critically appraise published research works focusing both on strengths novelties and weaknesses in the hypothesis formulation, on the methods and instruments used, on the discussion of the results. A critical analysis should enable nurses to start learn to read and eventually write a research protocol, possibly avoiding some common mistakes. PMID- 8715195 TI - [Evaluation of the risk of decubitus lesions in hospitalized patients]. PMID- 8715197 TI - [International observations. The era of limits. The rationing of health care]. PMID- 8715196 TI - [The good and the bad in prison]. PMID- 8715198 TI - Making a difference. A health delivery model for rural Saskatchewan. PMID- 8715199 TI - Making a difference. PMID- 8715200 TI - Making a difference. Bringing the wellness message to teens. PMID- 8715201 TI - Making a difference. Northern Saskatchewan: television drama occurs in real life. PMID- 8715202 TI - Making a difference. Facing the unknown: elected nurses take on new challenges. PMID- 8715203 TI - Making a difference. Aboriginal nurses and health professionals in high demand. PMID- 8715204 TI - Making a difference. Home-based nursing: making the shift from hospital to home. PMID- 8715205 TI - Making a difference. Made-in Saskatchewan guidelines for cholesterol testing and treatment. PMID- 8715208 TI - Nursing image still outdated. PMID- 8715207 TI - Florence Huey: a voice for nurses yesterday and today. PMID- 8715210 TI - Andrea Weipert. Interview by Angela Pearch. PMID- 8715211 TI - Psychiatric nursing. PMID- 8715213 TI - Eating disorders: a response to shame. PMID- 8715212 TI - Breast cancer: lessons from an epidemic. PMID- 8715214 TI - Women and stress. PMID- 8715215 TI - For victims of domestic abuse--a shattered world. PMID- 8715216 TI - We are family. PMID- 8715217 TI - [Quality in nursing]. AB - Different definitions of quality of health care were reviewed. These difinitions are considered to be the fulfillment of desired results in the health of individuals and overall population and some refer to the health definition itself and to the influences of the patient, family and health system. In our country the objectives of health care quality are implied in the definitions revised. Because of the importance given to the evaluation of quality for health policy definitions some methods of analysis for the structure, process, and results of the system are reported from the very beginning of their application. Factors which involve nursing care in the quality of health care are described taking into account that its representation at administrative levels of the Health Care System and its enhancement impels of promotes the development of nursing actions in the structure and process of the system related to the results. PMID- 8715218 TI - [The effect of breast feeding and its duration on the health of the infant (December 1992-February 1993)]. AB - A study was carried out from a sample of 300 infants born at "Mariana Grajales" Obstetric and Gynecological Teaching hospital, Santa Clara, from December, 1992 to February, 1993. Some factors which influence upon breast feeding were studied. A survey including personal identity data and others was applied. Factors aforementioned were analyzed and it was found that breast feeding was maintained during the first 4 months of life in 58.0% of infants. In addition, acute diarrheal and acute respiratory diseases were found to be more frequent in those infants breast fed during a shorter period of time. A significant relationship between maternal occupation and lactation was also confirmed. No relationship was observed between nutritional status and lactation, although the greatest percentage is found in infants with normal body weight with 38.0% of breast-fed infants and 25.3% of non-breast fed infants. PMID- 8715219 TI - [Health problems in the population of a dispensary in Cienfuegas]. AB - A study was performed on the entire population served by the medical office #36 from the teaching area V of Cienfuegos municipality with the aim of determining the main health problems and the effectiveness of measures designed to eliminate or improve such problems. Among the principal results are: the incorporation of elderly subjects to "Grandparents Clubs", the reduction of smoking and an improvement of sanitary education, although certain difficulties are still present regarding the latter due to increased intestinal parasitic infections. PMID- 8715220 TI - [Implantation of definitive pacemakers: experience with 500 patients]. AB - Five hundred patients to whom definitive cardiac pacemakers were implanted from June, 1978 to June, 1992 were studied. A predominance of males and the age group of 61-80 years was observed. The most frequent reason for seeking medical care was syncope, and III grade atrioventricular block is the most frequent cause requiring this kind of treatment whose etiology was associated with the sclerodegenerative disease of the conduction system. The right cephalic vein was the most used via of approach; among the complications recorded only 2 resulted in death of the patient (perforation of the left ventricle and traumatic pneumotorax.) PMID- 8715221 TI - [Artificial respiration in status asthmaticus. Five years experience in an intensive care unit]. AB - A restrospective study was performed on patients presenting with status asthmaticus and admitted to the Intensive Care Unit of the Provincial Clinical and Surgical Teaching hospital, Pinar del Rio, within a 5-year period and who underwent mechanical ventilation representing 32.65% of the overall number of admissions in the ICU. Fifty seven percent of the patients were women with an average age of 48 years. The mean intubation time was 60 hours. At admission, the patient's clinical condition was assessed, as well as the cause for intubation and ventilatory parameters that are being used up to date. Complications and the mortality rate which increased to 9.38% were also analyzed. All results were compared to the ones from other authors, as well as from the national and international literature. PMID- 8715222 TI - [Diagnostic methods used in breast neoplasms. Structure of the National Subprogram for Breast Cancer]. AB - Different diagnostic methods used for the screening of breast cancer such as clinical examination of breasts, self-examination and mammography are reported. A detailed explanation is given regarding reasons for the indication of mammography which include latent symptoms in non-palpable lesions and asymptomatic cases, as well as the advantages of this diagnostic method. The structure and design of the Subprogram for Early and Preclinical Diagnosis of Breast Cancer are explained and data on the situation of breast cancer in the country are reported. The goal of the survey applied and the structure of the mobile car is briefly presented, as well as the management of the different classifications such as low, mid, high and very high risk. The purpose of this paper is to describe the developmental and supporting actions undertaken by the health staff, especially by the nursing and health promotion staff. PMID- 8715223 TI - [Vaginal parasites and acute cervicitis: local treatment with propolis. Preliminary report]. AB - A prospective study is performed in patients who sought for medical care at 2 selected medical offices from "26 de Julio" Teaching Polyclinics, Playa municipality, during July 1st, 1991 and July 31, 1992. These patients presented with acute cervicitis and had vaginal smears with positive cultures of some kind of infection. A study group and a control group were formed and the study was double-blind according to a random sample created for that purpose. Vaginal dressings using 5% propolis were applied to the study group, while daily dressings with Lugol were applied to the control group for 10 consecutive days. A survey was designed and results collected. It is concluded that all patients from the study group did not present with any other symptoms after treatment was completed since negative results were attained in 100% of the smears and 90% achieved a total epithelization of the cervix within 10 days of treatment. The importance of nursing care for the success of treatment is highlighted. PMID- 8715224 TI - [Focus on risk in adolescent pregnancy]. AB - Results from the application of risk approach for the analysis of early maternity are reported. This analysis is made from the available medical records. Based on data obtained from the Registry of Deliveries of the Obstetric ward of "Madariaga de Posadas" hospital, Misiones, Argentina, a cross-sectional study of analytical observation was conducted. All records regarding deliveries occurring during the first semester of 1991, i,e, 507 cases were considered for the study and 3 characteristics were chosen: age, parity, and complications due to an inadequate prenatal monitoring. The first two characteristics, in combination, were used as an indicator of risk level and the third was crossed in a contingency table. The most significant findings were that 20% of 2 513 deliveries cared for within the aforementioned period of time corresponded to early maternity. Average age of the adolescents was 17 years ranging from 12-19 and standard deviation 1.5 years, modal parity was placed in the rank of 0. Of 507 deliveries at risk, 69 were considered high risk deliveries because of the age and 305 because of parity. Overall, 61 deliveries presented with complications due to inadequeate prenatal care. In general, it is concluded that the technique and the data source used are quite useful to quantitate the occurrence of early maternity and to design strategies for nursing intervention in the community. These women who receive medical care in the public hospital come from popular sectors and represent a well defined risk group requiring specific preventive medical care for the reproductive process as a whole. PMID- 8715225 TI - [Revision and considerations on the writing of scientific articles]. PMID- 8715226 TI - Watch your language! PMID- 8715227 TI - Salespeople and the Lactation Army: taking a stand for health and Human milk. PMID- 8715228 TI - Visual assessment of the baby's wide open mouth. PMID- 8715229 TI - Developmental and nutritional concerns relating to timing of supplemental infant foods. PMID- 8715230 TI - Timing of solid food introduction and safety of radiopharmaceuticals: agreement and disagreement. PMID- 8715231 TI - References: a conflict of scientific interest? PMID- 8715232 TI - Caution urged about assumptions relating to breastfeeding women in Africa. PMID- 8715233 TI - Does prenatal breastfeeding skills group education increase the effectiveness of a comprehensive breastfeeding promotion program? AB - A control/intervention study carried out in Santiago, Chile assessed the impact of five interventions on breastfeeding patterns and duration, and demonstrated a significant increase in full breastfeeding at six months (32 percent to 67 percent). Fifty-nine of 422 post-intervention women were included in a sixth intervention: prenatal group educational sessions emphasizing the skills necessary to initiate and maintain breastfeeding past the neonatal period. A significantly higher percentage of this subset of women were fully breastfeeding at six months compared to those who received only the five basic interventions (80 percent and 65 percent, respectively). The effect was greater among primiparous women. We conclude that prenatal group education with hands-on skills reinforcement is a significant and additive component of breastfeeding support, especially among those who have no previous breastfeeding experience. PMID- 8715234 TI - Estimating milk intake of hospitalized preterm infants who breastfeed. AB - This study describes the accuracy of clinical indices to estimate the milk intake of breastfed preterm infants. Twenty-nine mother-infant pairs were studied for a total of 39 breastfeeding sessions. Two NICU nurses (RN1 and RN2) and one certified lactation educator (CLE) collected data, each blind to the others' measures. RN1 and RN2 performed test-weights using the Smart Model 20 electronic scale. The CLE observed the breastfeeding and estimated milk intake, using clinical indices of milk transfer. Clinical indices did not provide an accurate estimate of milk intake (r = .48). The mean absolute difference between the test weights and clinical estimates was 5.79 ml, with a maximal difference of 20 ml. These differences were random in that clinical indices did not consistently over- or underestimate milk intake. None of 17 clinical indices of milk intake significantly lowered the magnitude of error in the clinical estimate. These data suggest that clinical indices cannot serve as a replacement for test-weighing of preterm infants when an accurate estimate of milk intake is necessary. PMID- 8715235 TI - The effect of exclusive breastfeeding on development and incidence of infection in infants. AB - Of 145 normal fullterm infants studied during the first year after birth, those exclusively breastfed for the first four months differed significantly from those not exclusively breastfed in physical and behavioral development, and resistance to infection. At four months of age, the mean weight of the exclusively breastfed group was higher (p < 0.05). At one year, the exclusively breastfed group showed more advanced. Personal-Social (p = 0.05) and gross Motor development (p < 0.05) on the Denver Developmental Screening TEst, and lesser cumulative incidence of infectious diseases (p < 0.05). This study demonstrated the beneficial effects of breastfeeding on development and resistance to infection in infants. PMID- 8715236 TI - Effects of storage, time, temperature, and composition of containers on biologic components of human milk. AB - Effects of steel and glass storage containers, duration of storage up to seven hours, and storage temperature on the concentration of humoral and cellular factors in 357 samples of human breastmilk were examined. Leukocytic functions were estimated by E-rosetting of sheep RBCs by T-lymphocytes and phagocytosis of S. cerevisieae by phagocytes. Immunoglobulin (Ig) levels were studied by single radial immunodiffusion technique (SRID). Viability and cell counts reduced during the storage period, but were greater than 60 percent of baseline values. IgG, IgA, and IgM levels in milk did not show significant decline after storage (p > 0.01). Milk stored in glass yielded a greater number of functional cells after storage at 4 degrees C. PMID- 8715237 TI - Breastfeeding in Belgium: tentative evaluation based on neonatal screening cards. AB - In Belgium as in most developed countries, the proportions of breastfed to bottle fed newborns is poorly known. A compilation of infant feeding information, available on cards which were completed at the time newborns were screened for inborn metabolic errors, indicates that in the years 1992 and 1993, 62.5 percent of infants were breastfed on day five of life. The data collected in this way were consistent for the years studied. Other countries may also be able to use screening cards to estimate breastfeeding initiation rates. PMID- 8715238 TI - An assessment tool for evaluating hospital breastfeeding policies and practices. AB - In 1991, UNICEF and WHO launched the Baby Friendly Hospital Initiative, a global initiative promoting breastfeeding within hospitals. This article describes the development and use of a detailed questionnaire to acquire baseline data on hospital breastfeeding policies in the southeastern Pennsylvania Delaware Valley and to classify hospitals by their level of implementation of the Baby Friendly Hospital Initiative. The purpose of the questionnaire is twofold: a) to describe and analyze hospitals' breastfeeding policies and practices, and b) to classify hospitals with respect to their level of implementation of each of the Ten Steps and the implementation of the Ten Steps overall. PMID- 8715239 TI - A breastfeeding evaluation and education tool. AB - New parents needs a way to assess the adequacy of their baby's breastfeeding. Health care providers, in addition, need a tool for teaching breastfeeding technique. This article describes a tool that focuses parental attention on eight elements relating to early breastfeeding: feedings, positioning, latch, suck, milk flow, intake, output, and infant weight gain. PMID- 8715240 TI - Twenty questions for the consumer: a quality assurance tool for the lactation consultant. AB - A simple post-service consumer survey is presented with which the lactation consultant can gather and analyze data to determine the extent to which services are meeting the needs of clients. The brief survey can be used to gather data from all clients or from a sample of clients. The survey can be administered by telephone or by postal return form. After analyzing the data obtained, the lactation consultant can identify where improvement in practice is needed. An additional " self assessment questionnaire" enables the lactation consultant to identify areas where personal growth is needed. PMID- 8715241 TI - Galactogogues: drugs to induce lactation. PMID- 8715242 TI - Possibilities for donor milk use in adult clinical settings--a largely unexplored area. PMID- 8715244 TI - Statistical report of the 1995 IBLCE examination. PMID- 8715243 TI - Percentage of fat in human milk: influence of the method of extraction. 1934. PMID- 8715245 TI - [Law for the Prevention of Risks at Work]. PMID- 8715247 TI - [The lateral surface of the sacral bone and the coccygeal bone]. PMID- 8715246 TI - [The necessity of learning in order to advance]. PMID- 8715248 TI - [Bacteriuria and pregnancy. Role of the midwife in primary care]. PMID- 8715249 TI - [Preventive measures. Accidental exposure to blood and body fluids (I). Risk associated with nursing activities]. PMID- 8715251 TI - [Discrepancy between nursing theory and nursing practice]. PMID- 8715250 TI - [Increasing the preoccupation with labor risks and protecting the health of the workers]. PMID- 8715252 TI - [History of nursing. Nurses in the 19th century. Academic-legislative developments]. PMID- 8715253 TI - [Hostile trait; cause of cardiovascular disorders and peptic ulcers]. PMID- 8715254 TI - [The proctoscope]. PMID- 8715255 TI - [Technical language in health care]. PMID- 8715256 TI - [Women's health: nurses are opening the way]. PMID- 8715257 TI - [Use of Cuban recombinant streptokinase. Nursing care]. AB - A study was performed on the application of thrombolytic agents during the first hours of the onset and diagnosis of a myocardial infarction with the aim of recovering coronary reperfusion and preserving the cardiac muscle. The therapeutic use of the Cuban recombinant streptokinase (Heberkinase) is analyzed in this paper. This is the fourth investigation conducted about streptokinase and its application in patients with a myocardial infarction. PMID- 8715258 TI - [Coronary risk factors related to ergometric and radiographic changes, September 1992 to May 1993]. AB - The population comprising all subjects over 15 years of age from a health area served by a medical office was analyzed by a survey with the aim of determining which patients were at risk of developing an ischemic cardiopathy and including them in the Coronary Disease Program implemented at the Cardio Center of Villa Clara. Nursing interventions in performing the different tests are reported. PMID- 8715259 TI - [Nursing care for patients with kidney transplants during the immediate postoperative period]. AB - Twenty medical records of patients who underwent a renal transplantation were reviewed with the aim of analyzing nursing care during the first 24 hours of the transplantation. The most significant aspects regarding these patients, as well as the most relevant actions undertaken with respect to nursing procedures were also analyzed. It is interesting to note that arterial hypertension is found to be among the leading causes of chronic renal failure. During the immediate postoperative period 5 patients required mechanical ventilation for more than 8 hours. Regarding diuresis surveillance and monitoring, 14 patients presented with an enhanced diuresis, but in 4 the urinary volume was lower than 50 mL/h. The need to treat those patients in an Intensive Care Unit by a highly trained and specialized nursing staff is highlighted. PMID- 8715260 TI - [Geriatric assessment]. AB - The geriatric assessment includes the evaluation of the patient's functional capacity, and it can be applied at any health care level, even at home. This allows to approach to the biological, psychological and social aspects, and facilitates the systematic observation of the therapeutic process and of the results from medical and social interventions with the aim of preventing, or at least, delaying the occurrence of functional disabilities. In addition, this evaluation allows to design an adequate programming of nursing care at any health care level, including the patient's home. PMID- 8715261 TI - [Program of primary care in school health. Historical report]. AB - The authors by means of this work report on the history of the Primary Health Care Program for School Children That was initiated in the 1980s in the city of Ribeirao Preto, Brazil. PMID- 8715263 TI - [Adequate programming of weight loss and liquid extraction during dialysis]. PMID- 8715262 TI - [Education in primary health care. Diagnostic evaluation of teaching using microcomputers]. AB - The use of microcomputers in medical education to assess the training process in Primary Health Care is analyzed. The experimental study consisted of 8 modules related to immunization campaigns, nursing care of healthy children, sanitation, nursing care of women, epidemiology and statistics, and nursing care of those diseases presenting the highest prevalence. The program is based on the basic software "Evaluation of Training", and designed in basic language. Each module consisted of a set of questions and each set had 3 different forms of feedback: the given form, the controlled form, and the form for searching. The independent variable of this investigation was the evaluation of training by microcomputers, and the dependent variable was the cognitive domain and development. The sample comprises 16 individuals studying on the eighth semester of a nursing course, and each group is made up by 8 students (a control group and an experimental group). Data obtained from this investigation were assessed by the test of nonparametric indicators. Results evidenced that the evaluation of training by a microcomputer provided a significantly higher performance in comparison with noncomputer assisted evaluation. Students were satisfied regarding the interaction with the microcomputer and with the performance of the evaluation. Further investigations in this field are recommended. PMID- 8715264 TI - Screen test: mobilizing the community against breast cancer. Interview by Michaleen Elabdi. PMID- 8715265 TI - Support for unemployed nurses. PMID- 8715267 TI - Increased direct access to services provided by registered nurses--need for new initiatives. PMID- 8715266 TI - Nurses take heart--your profession rates high in public awareness polls. PMID- 8715268 TI - Confronting unsafe practice issues in the workplace. PMID- 8715269 TI - Charting the future--"scope of nursing practice discussion paper". PMID- 8715270 TI - Expanded role for ENs. PMID- 8715271 TI - Industrial relations--the heart of the difference between the two major political parties. PMID- 8715272 TI - New York State to give mothers the right to HIV test results. PMID- 8715273 TI - In medias res: the ethicist as mediator. PMID- 8715274 TI - Ethics: a matter of judgment. PMID- 8715275 TI - Doing ethics at the bedside: reflections on the practice of ethics consultation. PMID- 8715276 TI - How abstract is my thinking as an ethicist in clinical settings. PMID- 8715277 TI - Whose story is it anyway? The roles of narratives in health care. PMID- 8715279 TI - Medicare managed care: the next frontier? PMID- 8715280 TI - [Reserved tasks in the health professions: the current and future legal status]. PMID- 8715278 TI - Beliefs and attitudes about cardiopulmonary resuscitation and DNR orders: a survey of attending physicians. PMID- 8715281 TI - [Reforms are necessary .... Interview by Karin Wagner]. PMID- 8715282 TI - [A common basis for continuing education]. PMID- 8715283 TI - [Barbers, hairdressers and people who took care of the sick]. PMID- 8715284 TI - [A decisive step forward.... Interview by Dr. Frank Weidner]. PMID- 8715285 TI - [Nursing and medicine in the Internet]. PMID- 8715286 TI - [A way to better utilization of patient resources]. PMID- 8715287 TI - [Dealing with feelings of disgust during nursing training]. PMID- 8715288 TI - [As a guest in Russia]. PMID- 8715289 TI - [The care of patients with AIDS]. PMID- 8715290 TI - [Children with burns. Surgical and orthopedic treatment]. PMID- 8715291 TI - [Children with burns. The story of Jessica]. PMID- 8715292 TI - [The nursing process]. PMID- 8715293 TI - [Nursing care. Bathing a child with burns]. PMID- 8715294 TI - [Children with burns. The necessary work]. PMID- 8715295 TI - [Reeducation of children with burns. Kinesitherapy and ergotherapy]. PMID- 8715296 TI - [Children with burns. Specialized care]. PMID- 8715297 TI - [Children with burns. Jessica's schooling]. PMID- 8715298 TI - [Social service. A link between hospital and family]. PMID- 8715299 TI - [Children with burns. The reconstruction work]. PMID- 8715300 TI - [Follow-up of children with burns]. PMID- 8715301 TI - [Drugs used in pediatrics. Pediatric therapy: practical utilization of drugs]. PMID- 8715302 TI - [Overweight]. PMID- 8715303 TI - [Children who have murdered and nursing ethics]. PMID- 8715304 TI - [Psychosomatic therapies]. PMID- 8715305 TI - [The place of psychosomatic therapy in psychiatric hospitals]. PMID- 8715306 TI - [Psychosomatic therapy in day care]. PMID- 8715307 TI - [Introducing psychosomatic therapies into day care centers]. PMID- 8715308 TI - [A promising first therapy]. PMID- 8715309 TI - [Being a therapist, one is a teacher]. PMID- 8715310 TI - [Killing father and mother. The tragic journey of a double parricide]. PMID- 8715311 TI - [Introducing a medico-psychological center into a rural area. Methodology]. PMID- 8715312 TI - [The stakes in nursing diagnosis for the pluridisciplinary team practicing in a medico-psychological center]. PMID- 8715313 TI - [An unjustified fear]. PMID- 8715314 TI - Understanding and empowering minority students in higher education. AB - A number of forces and trends confront America's educational institutions. None are as pervasive and encompassing as the issue of diversity which implies change in the daily operation of these universities. The change that must occur includes, but is not limited to, new policies that recognize and respect the tremendous ethnic and social diversity both on and off campus. Helping to fuel these changes are minority students on White campuses who are expected to participate in mainstream activities at the same time that they are encountering barriers and attitudes which stand in their way of doing so. This conflict exacerbates the attrition rate for minority students. This article address issues of diversity and change relative to minority students on predominantly White campuses. It provides insight for educational personnel on issues of diversity and change, ethnic identity development and development of minority student leaders in the broad collegial settings. By empowering minority leaders and organizations, institutions strengthen their ability to recruit, develop and retain minority students. PMID- 8715315 TI - Health promotion practices of smoking and non-smoking black women. AB - This descriptive study utilized Pender's Health Promoting Lifestyle Profile (HPLP) to survey a sample of Black women age 18 and older who attended three churches located in the southcentral United States. The prevalence of cigarette smoking is increasing among females, and Black females are at greater risk for complications related to smoking than are their White counterparts. The purpose of this study was to describe the health promotion practices of Black women who smoke cigarettes and those who do not smoke cigarettes. If differences were identified in the health promotion practices of these two groups, the findings would provide a basis for developing a plan for overall health promotion specifically tailored for Black women who smoke. The data analysis revealed no significant difference between the HPLP and subscale mean scores of the smokers and non-smokers. Scores were higher on a minimum of five subscales and the overall HPLP for women > 40, those with some college level education, or having annual household incomes of > $20,000, regardless of smoking status. MANOVA results revealed significant main effects of age on three subscales. Two-way ANOVA's revealed significant main effects of age and household income on HPLP scores. The results in this study indicate that education is the strongest predictor of health promoting behavior. Other demographic variables such as age and household income accounted for a variance in the subscale categories and overall HPLP scores. Nursing interventions with a health promotion focus must be developed with an emphasis on Black women who are younger than 40, those having a high school or lower educational level, and those in the lower socioeconomic class. PMID- 8715316 TI - Research methodology. Part IV: Understanding canonical correlation analysis. AB - Canonical correlation is presented as a technique to determine how sets of dependent variables are related with sets of independent variables. Canonical correlation reveals the strength of the relationship between the clusters using case data as illustration, three pairs of clusters (factors or profiles) emerged. Interpretation of the clusters are presented. As indicated in the case presentation, Canonical Correlation (CA) is the fourth in a series of methodologies selected for illustration as precursors to advanced statistics and modeling. In this paper, background will be given, a schematic example presented, sample size and CA, SPSS procedure to perform CA, and interpretation of CA and possible uses of CA in nursing research. PMID- 8715317 TI - Enjoying your work. AB - This article discusses eight strategies nurses can use to make the most out of work situations emotionally and to bring out the enjoyable in their work. The author stresses that enjoying your work can be an important aspect of unleashing energy and creativity and advises against separating work and fun for a healthy integration makes for a more healthy life. PMID- 8715318 TI - Ethical dilemmas. PMID- 8715319 TI - Technical and nursing roles in patient care for photorefractive surgery. AB - 1. The University of Ottawa Eye Institute is conducting clinical research protocols using the excimer laser. The state-of-the-art technology currently available ensures an ever-evolving role for the staff, and working as a team allows the patient the best possible care experience. 2. The role of the technologist (at Ottawa Eye Institute) includes ensuring the basic laser maintenance and calibration of the energy levels delivered by the laser; carrying out pre- and postoperative patient testing according to protocols; and assisting with the surgical procedure. 3. The role of the nurse (at Ottawa Eye Institute) includes ensuring the informed consent form has been signed and patient education been given. PMID- 8715320 TI - Ophthalmology's interaction with the Internet (you say, 'Internet what?'). AB - 1. You can be connected to the information Highway by a local phone call. So, you will have a monthly charge for your access, but not a more expensive phone bill. 2. Mail lists are discussion groups; actually, they are "computer remailers." When you join a mail list, your e-mail address is entered into a computer, along with a group of people with common interest on the Net. 3. The World Wide Web is the graphical and multimedia part of the Internet--color, sound, video, and all! To access the World Wide Web, you'll need a browser. A browser is a piece of software that allows you to look at all the Web's various home pages and websites. Organizations, individuals, universities, and corporations have Web addresses (called URLs) that allow you to look at their "roadside stand" on this Information Highway. PMID- 8715321 TI - Improving drop administration by patients. AB - 1. Patients that have difficulties with drop application may have poor compliance with therapy. We studied the effectiveness of teaching a technique of drop administration that allows the patient to steady the hand and observe the dropper tip during application. 2. Comparing the group taught the technique of application with another group who had their drops applied by a home caregiver, we found that those taught the technique were as reliable as the others in the delivery of their drops. 3. Although some patients will be physically incapable of applying drops, we found that the technique we describe in the present study enables effective and safe delivery of drops. PMID- 8715323 TI - Troubleshooting the postoperative IOL result. AB - 1. We all experience disappointment when a patient's postoperative visual result differs from the anticipated refraction. Although it is easy to initially place blame on an A-scan measurement, keratometer reading or an incorrect IOL calculation, our frustration may be tempered by applying a few basic troubleshooting steps. 2. There are several reasons for determining the cause for the discrepancy. First is to address the potential need for an IOL exchange if the refraction is dramatically unacceptable. Second, information gained from analysis of the first eye's results can aid in more accurately predicting an IOL power for the patient's fellow eye. A third reason is to learn from our mistakes to prevent their recurrence. A step-by-step analysis provides an unemotional method for reaching a much needed conclusion. 3. Ask yourself the following questions: Do pre- and postoperative measurements make sense and correlate with patient history? Are A-scan echo patterns and electronic caliper placement correct? Have correct values been entered into the formula? PMID- 8715322 TI - Vitamins for cataracts and macular degeneration. AB - 1. In some instances, the lay and scientific community already have concluded that antioxidants confer protection against cataracts and macular degeneration, and have been using them toward that end for some years. 2. Because vitamins are not considered drugs, their prescription and dosage are not subject to the regulation of the Food and Drug Administration. While some studies support their use, others do not. 3. Until sound randomized, controlled clinical trials have been completed in the use of antioxidant supplements and the development of cataracts and macular degeneration, the exact therapeutic role and dosage of antioxidants will remain less than clear. As is the case with any drug, short- and long-term side effects, as well as potential toxicity, have to be carefully weighed against the protective benefits in each individual case. PMID- 8715324 TI - There's a lot more to keratometry than just K-readings. PMID- 8715325 TI - [The nursing anamnesis. A concept for nursing assessment of the admission status in a surgical intensive care unit]. PMID- 8715326 TI - [Infections in surgical patients: etiology diagnosis, prevention. 1]. PMID- 8715327 TI - [Surgical concepts of continent bladder replacement]. PMID- 8715328 TI - [Pharmacotherapy in the aged]. PMID- 8715330 TI - [Satire. Male and female speaking behavior under the magnifying glass- introduction and warning]. PMID- 8715329 TI - [Therapy of defects in the oral mucosa in patients with hematologic or oncologic diseases]. PMID- 8715333 TI - President's message: mentorship: a key to promoting spinal cord injury leaders. PMID- 8715335 TI - The skeleton after spinal cord injury. Part 2: management of sublesional osteoporosis. PMID- 8715334 TI - Risk factors for pressure ulcers in the spinal cord injured in the community. AB - A retrospective review was performed on the medical records of 29 patients with traumatic spinal cord injury who were admitted for pressure ulcer research protocols within the Division of Plastic Surgery at a university medical center in Southeast Texas. The purpose of the study was to identify common factors in the community that may contribute to the development of pressure ulcers in the spinal cord injured person at home, as a prelude to development of a risk assessment scale for use by the home care nurse. Oot-Giromini's Web of Causation was used as the theoretical framework for this study. Findings support current pressure ulcer research literature, implicating a diverse array of physical and psychosocial elements such as nutrition, past history of pressure ulcers, and social support issues as risk factors. Further research is needed before a risk assessment scale is formulated. PMID- 8715336 TI - Collaborative efforts in achieving patient outcomes in central cord injury syndrome in the acute care setting. AB - Central cord injury results in motor impairment that is more prominent in the upper extremities than lower extremities with greater distal than proximal weakness. This altered health status directly impacts an individual's self care abilities. The combined focus of nursing care and occupational therapy delivered in a collaborative approach facilitates progress toward independence. An individual who has experienced central cord damage may be able to walk but not be able to eat, toilet, or dress without assistance. Most basic needs cannot be met independently without use of the arms. This type of injury has major implications for self care abilities. PMID- 8715337 TI - Designing patient education materials to increase independence. AB - The SCI nurse often has the responsibility to develop and utilize patient education materials related to various self-care routines and issues dealing with spinal cord injury that are comprehensive, understandable, and economical. The design and content of the material needs to be something that can be used as a teaching resource during the rehabilitation program as well as an information source after discharge. The SCI InfoSheets developed as a project of the Medical Rehabilitation Research and Training Center (RRTC) in Secondary Complications in Spinal Cord Injury at the University of Alabama at Birmingham can be used to facilitate the goals of the SCI nurse in patient education. PMID- 8715338 TI - AASCIN research needs assessment and interest survey: report to membership. PMID- 8715339 TI - [A plea for freedom of thought]. PMID- 8715340 TI - [The end of meekness. Men's fear of strong women]. PMID- 8715341 TI - [Nursing and environmental protection. VI. A look across the fence: alternatives in the disposition of of refuse]. PMID- 8715343 TI - [Alginates in septic surgery. Dressings can speed wound healing]. PMID- 8715342 TI - [Ambulatory surgery in the hospital. Legal and management aspects are not yet definitely settled]. PMID- 8715344 TI - [According to x-ray regulations, operating room personnel must have special training]. PMID- 8715345 TI - [A working group is establishing nursing standards. Nursing is organized on a common basis in all units]. PMID- 8715346 TI - [Care of patients with colostomies or ileostomies. Social isolation is prevented by comprehensive education]. PMID- 8715347 TI - [Ovations for Validation therapy. Naomi Feil's way to the understanding of the aged is newly convincing]. PMID- 8715348 TI - [Violence in nursing. Sensitize yourself by understanding your own aggression]. PMID- 8715349 TI - [Congress on "alternative healing" almost sold out. Gentle medicine on an up current]. PMID- 8715350 TI - [Organization in nursing. Use your time meaningfully]. PMID- 8715351 TI - [Occupational disease in nursing personnel. Psychological stress in increasing for nurses]. PMID- 8715352 TI - [Nursing education in Finland. Research and administration are emphasized in the courses]. PMID- 8715353 TI - [The adjustment to a new nursing organization could be an enrichment]. PMID- 8715354 TI - [The Board of German Assistance for the Aged fears a decrease of services. The position of those needing care is drastically deteriorating]. PMID- 8715355 TI - [Nurses state considerable shortcomings in wound care]. PMID- 8715356 TI - [HIV and AIDS. Legal and social aspects of care]. PMID- 8715357 TI - [Mouth care in intubated patients with nothing-by-mouth diet in an in tensive care unit]. AB - The objective of this study is to value the effectiveness of a care plan of mouth care and at the same time, compare the effects of the use of two different solutions: physiologic serum and non-diluted hexetidine (Oraldine). The sample studied was formed by 40 intubated patients or patients with an absolute oral diet who were in the Intensive Care Unit of the University Clinic. 50% of the patients underwent oral hygiene with physiological serum, following the established protocol and the other 50% were treated with non-diluted hexetidine with a frequency of five times a day. The mouth status was checked at the moment of admission and then daily: lips, tongue, gums, palate, presence of residua and saliva characteristics. Also, a series of factors which could alter the buccal integrity were registered: oxygenotherapy, orotraqueal intubation, anemia, plaquetopenia, medicines, etc. Bacteriological controls of traqueal secretion were performed periodically. From the analysis of the results we can conclude that the care plan established is efficient, although it is not enough for patients with coagulation alterations (plaquetopenia). From the solutions used for oral hygiene we can say that although there have not been significant differences, the physiologic serum is the chosen product, as it maintains the mouth status better and helps to keep the oral microbiota. PMID- 8715358 TI - [Care plan for patients in prone decubitus. An experience from practice]. AB - Offering a specific integral attention to patients with SDRA in prone decubitus positions makes us establish a performance plan with the aim to know the problems derived from the change in position, the time staying in prone decubitus and to standardize a care plan. We review the clinic records of the patients admitted in our unit from March '93 to March '95 who were positioned in prone decubitus. Taking as a base the nursing care model of V. Henderson and the taxonomy of NANDA, we analyse the needs which have been altered, and determine the nursing diagnosis, complications and most frequent interdependent problems establishing the aim to accomplish, planning the performance and rationalization. Five patients were positioned in prone decubitus before planning the performance and four more afterwards. All the patients tolerated SNG diet keeping a correct bowel transit. One patient showed an ulcera at frontal level. There were neither comeal ulceras nor alterations in the oral mucossa. The vascular accesses remained permeable. DP caused facial and periorbital edema in all the patients. We did not observe any increase in the amount of bronchial secretions. The eight patients who tolerated the change in position stayed in prone decubitus for an average of 77 hours, with a range of 10 to 216 hours. Four patients were discharged from the hospital, two of whom showed movility alterations, independently of the time staying in prone decubitus. We state explicitly the nursing care, determine five nursing diagnosis, one problem and seven interdependent complications. Establishing the nursing care from the experience and review of the records has allowed us to be more specific and objective. Standardizing the specific care plans makes the nursing care easier when dealing with real problems as well as with the care of complications derived from this situation. PMID- 8715359 TI - [Decubitus ulcers in intensive care units. Analysis and care]. AB - The fact that intensive care patients suffer from ulcera is a daily evidence which has a negative repercussion. We have analysed prospectively a sample of 215 patients to know the incidence, prevalence, levels, and placement of the decubit ulceras to observe whether there is an association between the variables age, sex, staying end, diagnosis, diabetes, risk level and postural changes and ulceration incidence. To do so, we have created a nursing care protocol for decubit ulceras to unify criteria and norm the performances. One out of every five I.C.U. patients suffers from ulcera and 30% of them show four or more ulceras, being the sacro and the heels the most usual places. There is an association between the patient's age, number of days staying in I.C.U. and diabetes and a higher incidence of ulceration. On the other hand, patients with politraumatisms diagnosis, infections and respiratory pathologies suffer from ulcera more than others. There is a clear association between the time of staying without postural changes and the incidence of ulceration. The same thing happens with the high risk stay. Our population is over 61% of I.C.U. stay in high risk, and its incidence of ulceration is 21%. Comparing both parametres we obtain an idea of the prevention which nursing professionals perform. PMID- 8715360 TI - [Calcium antagonists]. PMID- 8715361 TI - [Nursing facing the new propositions for hospital organization]. PMID- 8715362 TI - [45th World Peace Conference of Religionists in Italy]. PMID- 8715363 TI - [Japan Visiting Nursing Foundation has started activities to promote home health service]. PMID- 8715364 TI - [Hospital management and role of nursing profession. Nursing management--a strategic approach in practice settings]. PMID- 8715365 TI - [Hospital management and role of nursing profession. Nursing contribution to hospital management]. PMID- 8715366 TI - [Hospital management and role of nursing profession. Hospital management participation through nursing project teams]. PMID- 8715367 TI - [Hospital management and role of nursing profession. Quality care supported by successful hospital management]. PMID- 8715368 TI - [Hospital management and role of nursing profession. Nursing philosophy contributes to successful hospital management]. PMID- 8715370 TI - [Liberation of medical service--medical information service]. PMID- 8715369 TI - [Hospital management and role of nursing profession. Hospital management should learn from Japanese industrial management system]. PMID- 8715371 TI - [Life of elderly patients at Farnham Hospital in England]. PMID- 8715372 TI - [Government committee to deal with issues of society and with fewer children and increasing elderly population]. PMID- 8715373 TI - [Report of a hospital director of nursing--my concern for nursing staffing level under the new nursing service]. PMID- 8715374 TI - [History of Japanese nursing that I lived: nursing in the 1930s and JNA admission to the International Council of Nurses]. PMID- 8715375 TI - [Medical topics. Recent advances in colon cancer]. PMID- 8715376 TI - [SCOPE--radiography treatment]. PMID- 8715377 TI - [Socioeconomic analysis of nursing services--nursing personnel demand and supply estimate and preparation of nursing personnel]. PMID- 8715378 TI - [Specialist nurses in private practice--home infusion therapist]. PMID- 8715380 TI - [The nursing shortage and the 1990s: realities and remedies]. PMID- 8715379 TI - [New medical fee system and nursing service--additional nursing fee for night shift, etc]. PMID- 8715381 TI - [Proposition for ethical consideration in nursing research. Japanese Nursing Association Research Review Committee]. PMID- 8715382 TI - [Committee on issues of society with fewer children and increasing elderly population]. PMID- 8715383 TI - [Committee to support care and independence of elderly people]. PMID- 8715384 TI - ["Uterus Day" in Kagoshima Pref. to provide telephone counseling]. PMID- 8715386 TI - [Median longitudinal section of a finger of an adult]. PMID- 8715385 TI - [Computers and decubitus ulcers. DecubituSW. Application of a program]. PMID- 8715387 TI - [Vaccination of children. How to increase coverage. Experiences with the vaccination program in Andalusia]. PMID- 8715388 TI - [Nursing as scientific knowledge, as technological knowledge, as technical activity]. PMID- 8715389 TI - [Planning, methods and development of a course in basic cardiopulmonary resuscitation]. PMID- 8715390 TI - [Nurses can contribute to the quality of life for the elderly in various ways]. PMID- 8715391 TI - [Cardiopulmonary resuscitation. How much do nurses know?]. PMID- 8715392 TI - [Preventive measures. Accidental exposure to blood and body fluids. II. Risk associated with nursing activities]. PMID- 8715393 TI - [Back pain. How nurses can identify and prevent the problem]. PMID- 8715394 TI - [Public health and nursing. New implications]. PMID- 8715395 TI - [Communication with the postpartal patient. An instrument after Roy's Model]. PMID- 8715397 TI - [There is no such thing as "it was always like this" in nursing]. PMID- 8715396 TI - [The Bulau thoracic drainage system]. PMID- 8715398 TI - [Start to be politically active]. PMID- 8715399 TI - [Injections. Correct preparation means avoiding injury]. PMID- 8715400 TI - [The nursing concept is being transferred into practice. I. The care of patients with tumors carries special demands]. PMID- 8715401 TI - [The rediscovery of a neglected part of care. Palliative care is one of the main tasks in nursing]. PMID- 8715402 TI - [Economy and quality of care on review: index--an electronic data processing program helps to save]. PMID- 8715403 TI - [Nursing and environmental protection. V. The inner life of cellulose is assessed by laboratory tests]. PMID- 8715404 TI - [Continuing education in intensive care and anesthesia. Work satisfaction and patient-centered care are guaranteed]. PMID- 8715405 TI - [Neighborhood help in Weil der Stadt. A visiting service assists the dying]. PMID- 8715406 TI - [Confidentiality among hospital staff. Nurses have to keep patients' secrets]. PMID- 8715407 TI - [Occupational diseases in nurses. 2. Back and neck pains are foremost]. PMID- 8715408 TI - [The influenza epidemic of 1918. Nursing personnel was doubly involved]. PMID- 8715409 TI - [We decided on patient-oriented care--what has changed?]. PMID- 8715411 TI - [The teaching of key qualifications. Claims and reality]. PMID- 8715412 TI - [Teaching and learning geriatric care. A patient-oriented integrative model]. PMID- 8715413 TI - [Consequences of nursing insurance for education in geriatric nursing]. PMID- 8715414 TI - [Can the ability to cooperate be learned or taught? An integrative teaching concept for geriatric care]. PMID- 8715415 TI - [Cancer of the larynx. Diagnosis]. PMID- 8715416 TI - [Cancer of the larynx. Treatment and therapeutic indications]. PMID- 8715417 TI - [First consultation at the ORL. Admission of the patient]. PMID- 8715418 TI - [Laryngectomy. Preoperative endoscopic findings]. PMID- 8715419 TI - [Partial laryngectomy. Nursing care in the operating room]. PMID- 8715420 TI - [Partial laryngectomy. Hospital nursing care]. PMID- 8715421 TI - [Reconstructive laryngectomy. Kinesitherapy]. PMID- 8715422 TI - [Organ procurement and religion]. PMID- 8715424 TI - [She was a nurse and my student in writing class]. PMID- 8715423 TI - [The female perineum. Low pathways of approach]. PMID- 8715425 TI - [Proposed new qualification "psychiatric health social worker" and its implication for the nursing profession]. PMID- 8715426 TI - [Increased nursing fees in revised medical treatment remuneration system. Staffing arrangement to meet nursing care needs under new system]. PMID- 8715427 TI - [Increased nursing fees in revised medical treatment remuneration system. Staffing requirement in hospital caring for cerebral-neuro patients]. PMID- 8715428 TI - [Increased nursing fees in revised medical treatment remuneration system. Increased dependency on nursing auxiliaries]. PMID- 8715429 TI - [Increased nursing fees in revised medical treatment remuneration system. Part of nursing care costed as auxiliaries' fees]. PMID- 8715430 TI - [Increased nursing fees in revised medical treatment remuneration system. Efforts to go on with current staffing level]. PMID- 8715431 TI - [Increased nursing fees in revised medical treatment remuneration system. New system does not favor psychiatric hospitals]. PMID- 8715432 TI - [Increased nursing fees in revised medical treatment remuneration system. How to balance quality of care and cost effectiveness]. PMID- 8715433 TI - [Increased nursing fees in revised medical treatment remuneration system. Nursing fee increase necessary for quality care]. PMID- 8715434 TI - [Health policy on care of the elderly, home health service, care of confused elderly patients]. PMID- 8715435 TI - [Report of director of nursing: looking back on the year 1994]. PMID- 8715436 TI - [We admire nurses: we are making progress]. PMID- 8715437 TI - [Development of independent nursing practice: ET nurse practitioner]. PMID- 8715439 TI - [Care of patients with AIDS and nursing education]. PMID- 8715438 TI - [History of Japanese nursing that I lived: graduating from Keio University Hospital School of Nursing and entering midwifery school]. PMID- 8715440 TI - [Pain control during operation and postoperative quality of life of TAE patients]. PMID- 8715441 TI - Shifting financial burdens: consequences for patients. PMID- 8715442 TI - Barriers to care and treatment: the challenge of daily radiation therapy. PMID- 8715443 TI - Breast cancer and mammography: an American Cancer Society profile of Philadelphia and Montgomery Counties. AB - The American Cancer Society, Philadelphia Division, has completed a comprehensive profile of breast cancer and mammography in Philadelphia and Montgomery Counties. The profile consists of three segments: (1) data on incidence of breast cancer and trends in stage at diagnosis; (2) estimates of breast cancer screening practices based on surveys of the population; and (3) results from a survey of mammography providers assessing the region's capacity for and access to mammography. The profile demonstrated that the Philadelphia Division is already close to achieving the national American Cancer Society's goals for breast cancer detection for the year 2000 for the percent of women screened and percent of cancers diagnosed in early stages. New and more ambitious goals for the Division must be set. Although capacity for mammography is high, outreach programs by mammography providers represent only 1% to 2% of all mammograms performed. The profile has been the cornerstone of new programmatic initiatives for the Division. PMID- 8715444 TI - Portraits of cancer survivorship: a glimpse through the lens of survivors' eyes. AB - Cancer survivorship is a tumultuous experience of balancing the elation of surviving life-threatening illness with the demands of chronic health concerns and altered life meaning. Increased awareness of the needs of cancer survivors is enhanced by large studies of this growing population, but also by appreciation of individual stories of survivorship. "Portraits" of survival, in the form of personal narratives, contribute to our understanding of the experience of cancer. Select portraits from a study exploring quality of life in cancer survivorship are shared to convey themes of rendering an old life, transition to a different life, and living an altered life after cancer diagnosis. PMID- 8715445 TI - Quality of life and nutritional support in patients with cancer. AB - Nutritional support is often initiated in patients with cancer who are unable to meet their nutritional needs by the oral route. Much has been written about the effect of nutritional support on physiological outcomes in patients with cancer. However, less is known about the relationship between improvement of nutritional status and quality of life. Trends in the treatment of cancer highlight the need for an examination of home nutrition support and quality of life. The purpose of this article is to describe the state of knowledge about the relationship of home enteral and parenteral nutrition support and quality of life. Research exploring the dimensions of quality of life (physical functioning, psychological status, interpersonal relationships and social functioning, financial concerns, and symptoms, and complications of nutritional support) is presented. Implications for clinical practice and research are identified. The trend for increased numbers of patients on home nutrition support emphasizes the need to understand the patient's and family's experience in managing this treatment in the home. PMID- 8715446 TI - Improving organizational performance: an introduction to the 1995 Joint Commission on Accreditation of Healthcare Organizations Standard. AB - The 1995 Joint Commission on Accreditation of Healthcare Organizations Standards represent a significant change in philosophy. Specific expectations for each discipline and department are no longer listed; instead, functions critical to patient care are described. With this change comes a new approach to the accreditation process; it is now focused on evaluation of the organization's performance of each of these functions. In addition, the Joint Commission on Accreditation of Healthcare Organizations expects that improvement activities be undertaken to systematically measure and assess the level at which the organization carries out these functions. Quality assurance and quality assessment have been replaced by performance improvement. The specific 1995 Joint Commission on Accreditation of Healthcare Organizations requirement is that "the organization designs processes well and systematically measures, assesses, and improves its performance to improve patient health outcomes." This article present an overview of the new standard, Improving Organizational Performance, and discusses issues to consider in implementing the standard in relation to the care of patients with cancer. PMID- 8715447 TI - Children's experiences with mothers' early breast cancer. AB - This article describes children's experiences with mothers' early stage breast cancer, a neglected area of study. Of 55 families from a larger study, 12 families had children between the ages of 2 and 21 years living at home. Family interviews, including the children when possible, at diagnosis and four additional times over the first year, were transcribed and content-analyzed. Two major themes emerged: awareness (entailing the child's cognitive awareness and emotional/behavioral response) and dependence. The child's developmental level influenced his or her understanding and the nature of the demands on the child and family. Because of the dependency needs of preschoolers, a chief concern for their parents was child care. Parents thought preschoolers were too young to understand so they gave simple explanations. Minor behavior problems were noted. More information was shared with school-aged children who believed their mother's situation was serious. Although children's concerns decreased, they remained evident months later. Effects of stress were noted. Home and caregiving demands usually increased and often interrupted adolescent's moves to independence and created role confusion and increased tension. Implications for parents and health professionals about interacting with children are suggested. PMID- 8715448 TI - Financial resources for adults with cancer. PMID- 8715449 TI - Does an alliance between a hospital cancer center and a community-based agency work? PMID- 8715450 TI - A survey of older Americans to determine frequency and motivations for eating fast food. AB - Senior citizens were surveyed at 11 senior centers in two states to determine their use of fast food restaurants for food and socialization purposes. Six and 11 percent of respondents in New Jersey and Texas, respectively, eat in fast food restaurants at least once a week. One-fifth of respondents in both states frequent fast food restaurants so they do not have to cook. One-quarter of respondents cited economics as the reason for fast food restaurant patronage. The study provided evidence that seniors patronize fast food restaurants in large numbers, have strong reasons for doing so, and their patronage and motivations do not differ geographically. PMID- 8715451 TI - Fibre and nutrient intakes of chronic care elderly patients. AB - It has been previously suggested that chronic care elderly patients are at increased nutritional risk. However dietary intake studies have not completely supported this statement. To determine usual dietary intakes, 32 elderly patients, mean (+/- SEM) age 84 +/- 1 years, from two hospitals, had 3-day dietary intakes estimated or weighed and analyzed for nutrient composition. The group as a whole had low intakes of dietary fibre but intakes of other nutrients were equal to or greater than the Canadian recommendations. The distribution of macronutrients also met recommended guidelines. In general, this group of elderly patients appeared to be eating well, however, some individuals results suggest nutritional risk. PMID- 8715452 TI - Diabetic diets revisited--time for a change. PMID- 8715453 TI - [Video-thoracoscopy. Principles and indications]. PMID- 8715454 TI - [Video-thoracoscopy. Preparations in the hospitalization service]. PMID- 8715455 TI - [Thoracoscopy. Preparations in the operating room]. PMID- 8715456 TI - [Video-thoracoscopy. Postoperative care]. PMID- 8715458 TI - [Supervision of thoracic drainage]. PMID- 8715457 TI - [Pleurotomy. Placing a thoracic drain at the patient's bedside]. PMID- 8715459 TI - [Removal of a thoracic drain]. PMID- 8715460 TI - [Aponeurotomy of the lower extremity after revascularization]. PMID- 8715461 TI - [Surgical activity in health facilities. First study in France]. PMID- 8715462 TI - [The curare-like agents]. PMID- 8715463 TI - [Ways of fixation of the uterus. Genital prolapse and surgical treatment]. PMID- 8715465 TI - Hereditary breast cancer. AB - An overview is provided of the genetics of hereditary breast cancer, as well as the clinical aspects and the management of this disease. In September 1994 a new breast cancer gene, BRCA 2, was located and BRCA 1, discovered in 1990, was sequenced (Miki et al., 1994; Wooster et al., 1994). The implications of these discoveries are immense. Tests are now available to women who have a family history of breast cancer that can determine if they carry one of these defective genes. The genetic nurse specialist can play a huge role in counselling women who wish to have predictive testing and in advising them about screening and risk. PMID- 8715464 TI - Clinical incidence of lymphoedema in breast cancer patients in Jonkoping County, Sweden. AB - The clinical incidence of lymphoedema of the arm in breast cancer patients was studied before and after general mammography screening in the county of Jonkoping, Sweden. There was a significant decrease of incidence of lymphoedema from 15% in 1983 to 8% in 1988. Clinical and pathological therapy data have been analysed in relation to lymphoedema. Significant factors contributing to lymphoedema were the number of lymph nodes examined and the number of pathologically positive lymph nodes. A higher proportion of patients receiving post-operative radiotherapy to the axilla developed lymphoedema, compared with those with surgery alone. A comparison between the two groups is impossible due to the fact that post-operative radiotherapy was prescribed only to patients with pathologically positive lymph nodes. PMID- 8715466 TI - Improvement of supportive nursing care in cancer patients autografted with bone marrow and mobilized peripheral blood haematopoietic progenitors. AB - We report that in cancer patients a dramatic reduction in infection rate, days of isolation, oral mucositis and hospitalization due to high-dose chemotherapy is achievable by autografting with haematopoietic progenitor cells (CPCs) circulating in peripheral blood following cancer therapy with high-dose cyclophosphamide (HD-CTX) and administration of recombinant haematopoietic cytokines. Thirty patients (29 lymphomas, one breast cancer) were treated with total body irradiation and high-dose melphalan followed by either: (i) bone marrow transplant (Group A); (ii) bone marrow plus HD-CTX-mobilized CPC transplant (Group B); or (iii) bone marrow plus HD-CTX- and cytokine-mobilized CPC transplant (Group C). Nursing care load was remarkably higher in Group A patients compared to Group B and C patients, thus demonstrating clinical advantages of transplantation of HD-CTX-f and cytokine-mobilized CPCs. PMID- 8715467 TI - Developing a new method to record care at home for children with cancer: an example of research and practice collaboration in a regional paediatric oncology unit. AB - Academics need to address the facilitation of nurses in practice to research and develop the care given to patients if nursing is to move closer towards a research-based profession. This paper describes a project involving collaboration between an academic department of nursing and nurses working in clinical practice providing care at home for children with cancer. The project involved an audit of the nursing care records for the service, and the development of a new style of record-keeping which described and quantified complex nursing care. The authors discuss the method and results of the project, describing the benefits of research and practice collaboration. PMID- 8715468 TI - A descriptive study of a cancer support group. AB - Six cancer support groups in the West of Scotland were studied. All 31 patients attending these groups were interviewed. The majority of participants were married females, middle class, 50-65 years old and were long-time survivors. Fifty-two per cent stated that they were very satisfied with the support they received. A high proportion of patients reported that they did not receive any counselling during their treatment from their cancer specialists (90%) or at present from a professional counsellor (84%). the general perceived health and 'global quality of life' among participants were found to be moderately good or good. Concerns of patients were studied and they reported optimistic changes. Needs assessment indicated that most patients (52%) require symptom relief and family support. Most patient (32%) reported that they had no problem at that time while the rest identified difficulties in home duties, shopping and transportation, and financial problems. The study suggests there is value in encouraging cancer patients and their relatives to take part in existing cancer support groups. PMID- 8715470 TI - Clinical supervision: a plea for 'pit head time' in cancer nursing. AB - The concept of clinical supervision has received considerable attention in the nursing literature of late, enjoying favour not only with many practitioners and managers, but academics and policy makers alike. The following paper considers the utility of clinical supervision within the cancer nursing setting, concentrating particularly on the development of self-awareness and the facilitation of responsive, reflective, therapeutic relationships. The paper is predicated on the belief that clinical supervision not only provides a valuable means of peer review and support, for nurses both in the pre- and post registration spheres, but that structured process of systematic supervision sustains the necessary environment for the protection and enhancement of quality nursing interventions (Butterworth, 1992). The potential of clinical supervision, as regards the wider development of nursing as a practice discipline, is illustrated by the incorporation of an extensive literature review which explores the theoretical and practical imperatives pertaining to the notion of clinical supervision, not only in relation to the individual, but to cancer nursing and nursing per se. PMID- 8715469 TI - The experience and impact of contacting a cancer information service. AB - Our objective was to review the quality of service delivered by a national cancer information service--the British Association of Cancer United Patients (BACUP)- and to consider the implications for future policy and practice. We used a sample of callers over a 10-day period, responding to a structured postal questionnaire. The sample included patients, relatives, friends, and general public who called the service with any concern relating to cancer. Of the 406 invited to take part, 282 responded (69%). The questionnaire was designed to evaluate the callers' perceptions of the information they received, their perceptions of the nurse's communication skills, the emotional impact of the call, and their overall satisfaction with the service. The results indicate that over 90% of callers evaluated the information they received and the nurse's communication skills positively, and there was no difference between the two major groups of callers, i.e. patients and relatives/friends. Both groups reported that the call had a positive emotional impact on them and that they were satisfied with the service. Emotional impact was predicted by quality of information for patients and nurse's communication skills for friends/relatives; overall satisfaction was predicted by nurse's communication skills for patients and both measures for relatives/friends. We conclude that the telephone is an effective medium for conveying information about cancer, and BACUP is meeting its needs. The needs of patients and relatives/friends are different--principally information for patients and support for relatives/friends--and it may be possible to improve training still further by focusing on those differences. The main challenge for cancer information services is to meet the ever-increasing demand. PMID- 8715471 TI - An audit of palliative care in dementia. AB - A retrospective case note audit was conducted in order to determine the most prevalent symptoms in terminal dementia and to assess the palliation given. Seventeen case notes were audited. Pain and dyspnoea were the most common symptoms. The palliation and treatment of constipation and oral candidiasis was within current accepted practice. Palliation of other symptoms were inadequate compared to current accepted practice. There appears to be a reluctance to prescribe opiate analgesia, and when this was prescribed the doses were not modified to achieve full pain or symptomatic relief. Many patients were unable to take medication orally, but syringe drivers were not used. The conclusions include the need for education of both nursing and medical staff as to the current principles of palliative care. PMID- 8715472 TI - A 5-year review of a lymphoedema service. AB - Data for 714 patients referred during a 5-year period to a specialist lymphoedema service in Oxford, UK, have been recorded on a database. The annual number of new patients almost doubled from 103 to 195. In addition to initial assessments, there were over 1000 follow-up appointments per year. Most patients were seen as out-patients. Fifty-six per cent of the referrals were from Oxfordshire and the rest from further afield. The two largest groups referring patients to the service were general practitioners (43%) and the radiotherapy department (26%). Most referrals were female (84%), and the main cause of lymphoedema was cancer and cancer treatments (68%). After 2 years, only 15% of patients were still attending the clinic regularly. The number of patients receiving intensive treatment increased by almost 40%. These findings emphasize the need to target resources and expertise selectively in order to provide an optimal service to patients needing specialist help. PMID- 8715473 TI - Changes to the NCLEX-PN test plan. PMID- 8715474 TI - Menopause update: change is good. PMID- 8715475 TI - Diabetes education: what every LPN should know. PMID- 8715476 TI - Educated as an adult: a competency based practical nurse program. PMID- 8715477 TI - The best medicine. PMID- 8715478 TI - Mum died in a nursing home ... thank God. PMID- 8715479 TI - Bronchodilators in chronic obstructive pulmonary disease. PMID- 8715480 TI - Biotechnology: a new dimension to pharmaceutical care. PMID- 8715482 TI - [JNA's position on creation of "psychiatric social worker" as a new category of health worker]. PMID- 8715481 TI - [Tender touch of nurses gave me the power to recover in ICU]. PMID- 8715483 TI - [New directions for hospital nursing department. Nurses to do nursing work only]. PMID- 8715484 TI - [New directions for hospital nursing department. Better nursing practice through better education]. PMID- 8715485 TI - [New directions for hospital nursing department. Nursing's contribution to hospital management]. PMID- 8715486 TI - [New directions for hospital nursing department. Changing service to meet the changing society]. PMID- 8715487 TI - [New directions for hospital nursing department. Nursing service to meet with people's expectations]. PMID- 8715488 TI - [New directions for hospital nursing department. Keeping up with social changes]. PMID- 8715490 TI - [Creation of psychiatric social worker as a health profession]. PMID- 8715489 TI - [New directions for hospital nursing department. Round-table talk: study visit to Beth Israel Hospital in U.S]. PMID- 8715491 TI - [Workshop for nursing practicum instructors: seminar for nursing administrators; national nursing examination review committee]. PMID- 8715492 TI - [Report of a hospital director of nursing--recruitment and placement of new graduate nurses]. PMID- 8715493 TI - [Life of elderly patient at Farnham Hospital in England]. PMID- 8715494 TI - [History of Japanese nursing that I lived: early nursing leader]. PMID- 8715495 TI - [Medical topics. Operation on an AIDS patient and not knowing it]. PMID- 8715496 TI - [SCOPE--hydrocolloid dressing for wound care]. PMID- 8715497 TI - [Socioeconomic analysis of nursing service--socioeconomic changes surrounding nursing]. PMID- 8715498 TI - [New medical fee system and nursing service--role of nursing in promoting home health service]. PMID- 8715499 TI - [Specialist nurses in private practice--ET nurse]. PMID- 8715500 TI - [AIDS nursing and nursing education in France--private practice]. PMID- 8715501 TI - [The nursing shortage and the 1990's: realities and remedies]. PMID- 8715502 TI - [Regulation amendment of death certificate]. PMID- 8715503 TI - [How gargling improves mouth problems]. PMID- 8715504 TI - [Evaluation of the nursing care for children 0-2 years old in a health center]. AB - The study is an exploratory-descriptive research that has as a main goal to evaluate the nursing consultation to 0 to 2 years old children in a health center. To do that the authors analysed the data collected in the children records from 1985 to 1988 and from interviews with mothers whose children had the nursing consultation in that time. PMID- 8715505 TI - [Women and nurses in domestic/family surroundings and in public. A theoretical and contextual approach]. AB - The study deals with the historical evolution of the woman in the society particularly in the familiar and public aspects. The intention to establish the relation, over the years, between the woman role and the nursing, a predominant female profession. PMID- 8715506 TI - [Prejudices and traditions in nursing]. AB - The authors try to identify the prejudices and traditions in nursing, thinking about the maintenance of the myths, stigmas and stereotypes throughout its history, facing the GADAMER's ideas. PMID- 8715507 TI - [Buying and distribution of drugs: perceptions of officers of a network of primary health care in the interior of the state of Sao Paulo]. AB - The study present analyse the process to buy and distribution of medicaments for the Basic Unit of Health in municipal district of state Sao Paulo. To achieve some general considerations about the National Politic of Medicaments in Brazil, to emphasize feature relative the its structuration in the Unique System of Health. PMID- 8715509 TI - [Qualitative approach: a contribution to nursing]. AB - This paper focus on some characteristics of the qualitative methodology. Some of these methods are explored such as: participatory research, phenomenology, grounded theory and ethnography critical theory Perspectives of their utilization in nursing research are examined. PMID- 8715508 TI - [Communicable diseases and isolation: a nursing student's perceptions]. AB - The Communicable Diseases have been associated to stigma and segregation for a long time. This investigation was carried out in order to detect the undergraduation students' perception concerning communicable diseases and isolation, as well as their expectation in working at isolation units. A questionnaire was elaborated and then validated by 3 referees. We attested that some of these students have stereotyped and negative conotations concerning the bearer of such diseases and isolation. Just a few of them mentioned that they would choose this specific unit as the workplace. It is important for us, professors, to know their previous knowledgement about the concepts of communicable diseases and isolation, so that we can reinforce the positive features and work with the negative ones during the courses. It can contribute to prepare these students to give an effective nursing care without fears and stigmas. PMID- 8715510 TI - [Elaboration of the concept of "Systematization of care": a stage in the consolidation of nursing intervention]. AB - The study analyse the implementation process of a discipline in Graduation Course which principal interest is "systematization of care to hospitalized adult". PMID- 8715511 TI - [Posture taken by members of the nursing staff during blood collection, intravenous drug administration and serotherapy]. AB - The members of the Nursing Team are submitted, many times, to posture attacks either due to the own requirement of the action or the utilization of improper corporeal posture during its execution. The present study aims at inquiring the type, length, frequency and posture changes adopted by the members of the Nursing Team during the execution of blood collection techniques, administering of intravenous medication and serotherapy. Through direct observation were recorded 10 activities corresponding to each one of the mentioned techniques, realized in attendance of patients from a University Hospital internment unit. The results evidenced that blood collection activity was 50, 35% effectuated in the inclined standing position, followed by the standing straight position, 49, 64%. Intravenous medication administering was executed with frequent adoption of the standing straight posture, 51, 09% (in special with the arm in frontal expansion), 45, 74% in the inclined standing position and 3, 15% seated. Serotherapy technique was executed with predominant adoption of standing position (86, 88%) in special with the arms in frontal expansion and 13, 11% in the inclined standing position. The unnecessary adoption, many times, of inclined standing posture indicates that Nursing personnel have been making possible the happening of spinal column attacks mainly due to the bad utilization of corporeal mechanic. The authors suggest more attention to these aspects, mainly at Nursing courses. PMID- 8715512 TI - Why the issue of patient information is important. PMID- 8715513 TI - Issues that influence information-giving in Europe. PMID- 8715514 TI - Patients' rights to information: the legislative framework. PMID- 8715515 TI - Dora's story. Dora Paraskevopoulou, president of the Hellenic Society of Mastectomy, Greece. PMID- 8715516 TI - The special case of clinical trials. PMID- 8715517 TI - Patient information as a necessary therapeutic intervention. PMID- 8715518 TI - Where do we go from here? PMID- 8715519 TI - Calling all researchers. PMID- 8715520 TI - An oasis of health for poor or homeless people who are HIV-positive. PMID- 8715521 TI - Independent practice in the outback. PMID- 8715522 TI - Looking out for family caregivers. AB - Nurses are regularly advised to become involved in community development initiatives, but what exactly does this entail and what are the precautions that should be considered? A 10-week, government sponsored, community-development program for family caregivers allowed nurses to offer education and support to six under-serviced areas of B.C. They offer a critique of the program and recommendations for the future. PMID- 8715523 TI - Debt profiles of new grads. AB - The economic climate in Ontario in particular and Canada in general (especially, severe problems with government budget deficits and accumulated long-term debt) has led to increasing pressures to off-load financial responsibilities for postsecondary education from governments to those who reap the primary benefits of such education--namely, the students. As a result, there have been sizable increases in tuition fees for postsecondary programs in recent years. It is reasonable to assume that cash-strapped governments will continue to expect students in university and community college programs to pay an increasing share of their education. In fact, this is one of the major options set out in the recent federal government paper for reform of social programs. PMID- 8715524 TI - Who cares for the caregivers? AB - Like the rest of Canada, the city of Ottawa is facing the "age wave"--a rapidly growing on-rush in the number of seniors. Indeed, Ottawa-Carleton has one of the fastest growing senior populations in the province of Ontario. About 70,000 of the residents, or 11 per cent of the population, are over 65, and this number is expected to increase to 15 per cent by the year 2011. PMID- 8715525 TI - [Reconsidering the approach to native peoples]. AB - Reconsidering nursing interventions among native people. The object of this study was to develop appropriate material to illuminate ways of enhancing interactions between native clients and clinical nurses. The chosen site was Roberval Hospital in Quebec, which serves the western and northern shores of Lac-Saint-Jean as well as the Chibougamau-Chapais region. A survey, using both a questionnaire and semi structured interviews, was conducted among a sample of 48 nurses working at that hospital and 29 native clients representing three distinct generations of the Montagnais people residing on an Indian reserve located eight miles from the hospital. Results underline that various cultural elements are at play when nurses and native people interact with each other. On the one hand, natives view the main problems as relating to communication, definition of values and family dynamics. For most nurses, communication is the main stumbling block. They readily recognize that their initial training has not prepared them to maintain effective relationships with native people. To enhance nursing competency and smooth over cultural barriers, the authors recommend appropriate transcultural training for nurses. Meanwhile, native people would benefit from information sessions on the functions of various health professionals in a hospital setting as well as the role and organizational structure of such an institution. PMID- 8715526 TI - Release of confidential information to the police. PMID- 8715527 TI - Mandatory practice hours. PMID- 8715528 TI - Balancing act. PMID- 8715529 TI - The growing importance of health research. PMID- 8715530 TI - Charter marks the way for children's services. PMID- 8715531 TI - How to write an essay. PMID- 8715532 TI - Child health in Moscow. PMID- 8715533 TI - Evaluation of a home care renal nursing service. PMID- 8715534 TI - Peripheral stem cell transplants in children. PMID- 8715535 TI - Nurse stress in cancer and leukaemia care. PMID- 8715536 TI - Reflective practice (continuing education credit). PMID- 8715537 TI - [Introductory report]. PMID- 8715538 TI - [Identity and specificity in nursing]. PMID- 8715539 TI - [Professional responsibility and autonomy]. PMID- 8715540 TI - [The role of nursing education: analysis of new education courses, proposals for improvement and development]. PMID- 8715541 TI - [Nursing education in the Apulia region]. PMID- 8715542 TI - [Possible consequences of nursing care in response to personal care needs]. PMID- 8715543 TI - [Possible effects of the organization and management of the nursing services on nursing care]. PMID- 8715544 TI - [Possible effects of basic, specialist and superior training on nursing care]. PMID- 8715545 TI - [Possible effects of nursing research]. PMID- 8715546 TI - [The quality of nursing care]. PMID- 8715547 TI - [Evaluation of the quality of nursing care: principles and objectives]. PMID- 8715548 TI - [Norms and criteria for the evaluation of the quality of nursing care]. PMID- 8715549 TI - [Determination of the work load for the evaluation of the quality of nursing care]. PMID- 8715550 TI - [Importance of nursing rules consistent with ethical disciplinary values]. PMID- 8715551 TI - Ethical, social and legal issues in Huntington disease: the nurse's role. AB - The nurse's role will be discussed in relation to the issues which may present as the result of our ability to use predictive tests for neurodegenerative disease. Huntington disease is an autosomal dominant inherited disease, characterised by emotional problems, abnormalities of movement and dementia. The disease is slowly progressive leading to a severely debilitated state and finally death in ten to twenty years. In 1983, DNA testing became available for persons at risk for Huntington disease and for confirmation of diagnosis for those showing symptoms. The availability of testing presents many ethical, social and legal issues for persons at risk, health care professionals and other segments of society. This paper will briefly review the genetic transmission and profession of Huntington disease. It will outline some of the benefits as well as some of the risks and problems DNA testing presents. PMID- 8715552 TI - Progressive supranuclear palsy: managing the disabilities and providing nursing support. AB - Progressive Supranuclear Palsy is an uncommon progressive neurodegenerative disorder which may initially present as Parkinson's disease. It is characterized by an abnormality of voluntary eye movement, pseudobulbar palsy, axial dystonia/rigidity, postural instability, mental changes and bradykinesia. The symptoms may be present for up to 4.5 years before diagnosis with an average survival rate after diagnosis of about two years. However, the diagnosis is often make much sooner when the individual is assessed in a specialty Movement Disorder Clinic. At the time the diagnosis is made the individual and family will have no awareness of the devastating impact this illness will have on their lives. Often they do not understand the mechanisms of deterioration, nevertheless, they must learn how to manage the severe disabilities over a short time period. This article will address the symptomatology of Progressive Supranuclear Palsy, how it differs from Parkinson's disease, the pathophysiology, treatment, recommendations for managing the disabilities, providing nursing support, care for the caregiver, future care planning, the legal options of Enduring Power of Attorney, Trusteeship, Guardianship, Advance Directives, and consideration for autopsy. PMID- 8715554 TI - [Breast feeding--no thank you! Or maybe?]. PMID- 8715555 TI - [Review--preview]. PMID- 8715553 TI - A self-care assessment tool (SCAT) for persons with a spinal cord injury: an expanded abstract. AB - The purpose of this study was to develop a reliable and valid tool that assesses the cognitive and functional skills needed for self-care in persons with spinal cord injury (SCI). The Self-Care Assessment Tool (SCAT) assesses cognitive and functional skills in eight self-care areas: bathing/grooming, nutritional management, medications, mobility/transfers/safety, skin management, bladder management, bowel management and dressing. The tool was carefully developed and has demonstrated content validity. Using two samples of veterans seen in the SCI clinics of two Southern Veterans Affairs Medical Centers, n = 13 and n = 15, interrater reliabilities computed by Pearson product moment correlations for the cognitive, functional and total scores ranged from .69 to .94. Test-retest reliabilities using Pearson product moment correlations for the cognitive, functional and total scores of two groups (n = 14 and n = 15) ranged from -.06 to .86. Regarding predictive validity, R2 was found to be .61 to .90 for the cognitive, functional and total scores. Although continued reliability and validity studies are needed, the SCAT has potential to measure patient rehabilitation outcomes, to evaluate nursing care approaches and to serve as a quality assurance indicator for nursing care. PMID- 8715556 TI - [To be or not to be. Questions about the future of health care. Attempt at a critical analysis of the system from the viewpoint of nursing, accompanied by visions for the future of the profession]. PMID- 8715557 TI - [Motives for choosing the nursing profession in the 19. and 20. centuries]. PMID- 8715558 TI - [Patients' expectations from women in nursing]. PMID- 8715559 TI - An argument for outsourcing. PMID- 8715560 TI - Campaigning for the working skin. PMID- 8715561 TI - Smoke gets in your eyes. PMID- 8715562 TI - Trike: a cause for concern? PMID- 8715563 TI - Medico-legal questions. PMID- 8715564 TI - Workplace continence awareness. PMID- 8715566 TI - [Changes of nursing paradigms. 1. Martha Rogers' theory]. AB - Change of paradigm in nursing. Part 1: Theory of Martha Rogers This article is an attempt to describe changes which are taking place in nursing. These changes are explained within the framework of Martha Rogers' theoretical basis of nursing. As a first step theoretical concepts are defined and three paradigmes identified by Margaret Newman are being explored. As a next step the scientific basis of nursing as defined by Martha E. Rogers is summarized. Elizabeth A.M. Barrett's "Theory of Power" and theoretical concepts of Margaret Newman are used to explain the practical relevance of Rogers' theory. Examples of Rogerian science-based nursing practice conclude this article. PMID- 8715567 TI - [Learning from mistakes]. AB - Learning from mistakes. Moral aspects in nursing Medication errors are experiences which may serve to demonstrate variables of moral action in nursing. Qualitative research in nursing is asking philosophical questions by exploring the meaning of experiences. Still, philosophical answers cannot be expected from the use of research tools that have been developed by the social sciences. The usefulness of such research is found foremost in the thorough understanding of the problems relating to a certain topic and in making complex realities more transparent by analysing concrete data. Recent feminist philosophy emphasises thinking in the context of empathy and concrete relationships. Ethics, taking recourse to these basic elements will be contextual. In this paper I offer some results of discourse analysis as a qualitative research approach to the understanding of the experiences of nurses who have been involved with medication errors. The interpretation of narrative texts allows a close look at everyday nursing experiences. Herewith, a contribution is made to a contextual ethics that is of value to nursing. I show how different modalities of identification are likewise compatible with a teleological and a deontological approach to ethics. Even though an ethic of care may well dominate human relationships, such an ethic does not have to dispense with rules and policies. PMID- 8715568 TI - [Moral action in nursing. Considerations from an ethical viewpoint]. AB - Moral action in nursing: reflections from an ethical point of view Does nursing need specific ethics? This question will be answered in two stages. First the history of how the profession came into being will be summarised. In the second stage the opportunities and the limitations will be touched on, regarding the shape of the profession. A short presentation of Kohlberg's theory is the starting point for an attempt to reconstruct the origins of moral competence. How far nursing will accept this is questioned. Ambivalence in the ideology of the caring-relationship is examined. PMID- 8715569 TI - [A scientific discourse on theories of morals and ethics]. AB - An academic discourse on ethical and moral theory The papers in this issue of Pflege were delivered by the authors at a conference on Moral Action, held in Osnabruck on Nov. 11th 1994. In the argument which follows, Marianne Arndt's critique points to the fact that Bondolfi's paper is based mainly on Kohlberg's theory and offers, in contrast, research by Gilligan. Bondolfi stresses that men and women are equally capable of rational and emotionally motivated action. Being "human" is a characteristic of members of both genders. PMID- 8715570 TI - [To what extent do patients with coronary heart disease want to be informed?]. AB - To what extent do patients suffering from coronary heart disease want to be informed? New legislation on patients' rights, passed in Austria in 1993, laid down what information patients are entitled to received. This raises dilemmas. On the one hand the patient has the right to be fully informed about his condition, on the other hand, disclosure could cause sufficient stress to impair the prognosis. The doctor must balance his duty to inform against the therapeutic reasons not to inform, in his duty to care. The empirical part of this research is concerned with the wish to be informed, of patients with coronary heart disease. A questionnaire was administered to patients after bypass operation. It is hoped that the results will be helpful to doctors. Nurses, who are not legally entitled to give information may, nevertheless, find it helpful to know about patients' individual needs for information, when they deliver professional nursing care. PMID- 8715571 TI - [The challenge of the health sciences to nursing]. AB - Health science, a challenge to nurses! We live in an age of old people who are chronically ill and in need of nursing care. Modern medicine's reply to the panorama of sickness and demography is to renew attempts to conquer the somatic progression with technology and chemistry. Will nursing be sucked into these developments or will it tackle new duties, arising, for example, in health promotion and rehabilitation? There are also problems of reshaping the health care systems. Nursing will have to be involved in this because of special professional experience and the developing nursing sciences. This contribution deals with the human face of medicine and nursing and touches on proposals for a patient-oriented redesign of the health-care system. PMID- 8715572 TI - [What kind of teachers are needed for nursing? Aspects of an educational concept specific for nurses]. AB - What kind of teachers does nursing need? Issues specific to nursing education Conditions which pertain to the future of education are discussed first. It is also necessary to look at reforms in nursing and in nursing education which have become necessary, and at the importance of the nursing sciences to the educational system and to the structure of the profession. One has to examine the function of nursing science in linking theory and practice. The author believes that only then can one examine the issue of educating nurse teachers. PMID- 8715573 TI - [Professionalism in nursing education--considerations on university education for nursing teachers]. AB - Professionalism in nursing education. Thoughts about higher education for nurse teachers The current discussion about higher education for teachers of nursing contains various aspects such as those belonging to education policy, public health or didactics of higher education. The first step will be a report on the situation and the second step reintroduces a conception of teacher training. The first part of this essay contains politico-educational and educational considerations as well as those refering to a theory of professionalization. It gives a description of the alternative of college (Fachhochschule) or university training for teachers of nursing and its consequences on the training of nurses. The second part presents a conception for teachers training, which results from educational theory and refers to the theory of professionalization: the training process taken as a autopoetical process founded on relation. PMID- 8715574 TI - [Motives of the career choice and expectations of the profession in nursing students]. AB - Motivation in the choice of a career. What expectations do students-to-be have of nursing? Various groups of nursing students (n = 267) were questioned about reasons for their choice of career, and comparisons were made. In contrast to earlier research findings, there was a multitude of motives. One can no longer assume that nursing students are a homogeneous group. The intrinsic motivation, that of wanting to be useful, is still high, as in previous investigations. Extrinsic reasons (s.a. pay, conditions of work, etc.) which were previously hardly mentioned at all, have increasing importance. A further new reason given for the choice of the career is the opportunity it offers for personal development. It is no longer enough to assume that for future students the motivation of wanting to be useful can be taken for granted, and that all that is necessary for maintaining it, are satisfactory working conditions. In order to ensure job satisfaction and to prevent frustration it will be essential to create conditions which ensure that the motivation to be useful remains intact but which also promote personal growth and development. PMID- 8715575 TI - [Nursing and power--use of the so-called poststructuralism theory for the analysis of the power-relationships in the "female" nursing professional]. AB - Nursing and power. Poststructuralism and its application to the power relationships in the "female" occupation of nursing The theory of Foucault is used to outline a "genealogy" of nursing, historically a female occupation, subordinate to medicine. Foucault's ideas about power, discourse and panopticon are analysed in relation to nursing. The concept of discourse enables the writer to see the "feminine ideology" of society as the effect of power on the identity, behaviour and expression of emotion of women in nursing. The hospital and the "Mutterhaus" can be understood as panoptical institutions. Hospital nurses are in a position in which they have powers of disciplining patients and at the same time are themselves subject to surveillance and normalisation. As a concrete example, the techniques of using power, which originate from monasteries, form "norm-traps" when applied to nursing. Finally, not only the question of power, but also that of resistance to power, is addressed. Professionalisation and the move towards science-based nursing can be seen as "counter discourse". This is meant to set up the nurses' own knowledge base, their understanding of the nurse patient relationship and of nursing practice, against the view of nursing as a profession subordinate to medicine. Foucault's view of power and resistance leads the writer to align herself with "the other side", in this case professionalisation in nursing. But at the same time one is invited to take the offensive in the debate of old and new contradictions. PMID- 8715576 TI - NACLNC Conference--a laboratory in strategic alliances. PMID- 8715577 TI - Weaving a home on the World Wide Web. PMID- 8715578 TI - On-line services can make or break your on-line experience. PMID- 8715579 TI - Indigenous nurses. PMID- 8715581 TI - Lights, camera ... nursing! Nursing in focus. PMID- 8715580 TI - Clinical education: a system under a pressure. PMID- 8715582 TI - Nursing and the law. Infection control and the role of nursing care plans. PMID- 8715583 TI - An adult survivor's story. PMID- 8715584 TI - Psychiatric care: change means opportunity. PMID- 8715585 TI - Nurse educators visit former Soviet Republic. Interview by Verna A Rhodes. PMID- 8715586 TI - [Reorganization of the graduate course in pediatric nursing at the nursing school at the Federal University at Minas Gerais. Evaluation of the process]. AB - In this paper, the faculty from the Pediatric Nursing Subject (Undergraduate Course) at the Nursing School of Universidade Federal de Minas Gerais evaluate a teaching project. Its main purpose was to reorganize the Subject based on ABATT; MCMAHON's approach to the teaching-learning process. The authours evaluate the results and highlight the positive aspects occurred due to the reorganization and confirm the need to continue the process. PMID- 8715587 TI - [Effects of Cartesian and emergent paradigms on the approach to health care]. AB - It deals with the approach to the health-disease and patient process, according to the Cartesian and emergent paradigms. It is taken into consideration that, due to the influences of the new paradigm, that is, the emergent one, the professionals of health, including nurses, are adopting more and more the holistic approach in the treatment of their patients. PMID- 8715588 TI - [Nutritional and disease diagnosis in institutionalized children. Day care center Elisio Teixeira Leite, Perus, SP 1993]. AB - As a part of one survey about health conditions in institutionalized children, the nutritional status were evaluated. Data were obtained through antropometric assessment in a sample of 111 pre-school children matriculated in a small official day-care center in Perus-SP. According to Gomez, 50.0% of all that children were classified as malnourished, 23.4% of them in the I degree; 22.4% in II degree and only 4.3% in III degree. Taking into account the Waterlow's classification there was a highest frequency of cronic-actual malnutrition (30.8%) concentrated in children between 12-48 months of age. Additionally, these children presented highest incidence of infective-respiratory and intestinal diseases. These evidences lead on to the improvement of the follow-up of children growth and food consumption, specially that under 24 meses, to attempt for an early identification of malnutrition, thus increasing the chance of nutritional recuperation. PMID- 8715589 TI - [Acute myocardial infarct: orientation manual for patients]. AB - The authors present a booklet for the patients with acute myocardial infarction. The main purpose of the booklet is to be a complement in the treatment for myocardial infarction patients, the leading cause of death in Brazil. It contains information about the disease, risk factors, return to work and treatment. PMID- 8715590 TI - [Nursing diagnosis of sexual dysfunction. Defining characteristics in patients with spinal cord injuries]. AB - The alterations of sexual function after spinal cord injury demands assistance because of cultural, psychological and biological aspects to keep. This study shows the defining characteristic, relacioned to nursing diagnosis sexual dysfunction (NANDA), in spinal cord injured patients on rehabilitation program. PMID- 8715591 TI - [Social roles of hospitalized patients]. AB - The research brings emotions, behaviors and feelings analysis of the nurses, patients and parents relationship that occurs during the hospitalization, and shows the consequence of this for all involved persons. PMID- 8715592 TI - [Nursing process in the ambulatory care of adults with arterial hypertension: concepts and contradictions]. AB - This study deals with the health assistance phenomena in general, and the nursing assistance in particular, as it occurs in Hypertension League. Its main theoretical framework, supported by the historical and dialectical materialism, it the labour process theory and its elements: Finality, Object and Means/Instruments. The investigation reveals that the workers conceptions about their labour process express a reiterative, non-creative and non-reflexive praxis. The labour process of nursing assistance was reiterated with Mean to the medical labour process, and to be seem emptied of the an scientific knowledge. PMID- 8715593 TI - [Care of mothers of newborns in intensive care units: experiences, feelings and expectations of the mothers]. AB - The purpose of the study was to examine the experiences, feelings and expectation of mothers of high risk newborns. The population was a group of 20 mothers of high risk newborns of three hospitals in the City of Sao Paulo. Interview with the mothers was the method of data collection containing opened and structured questions. It was verified that most of the mothers had none or only a little interaction with the newborn after delivery; the eye contact was the most referred during the staying of the newborn in the Intensive Care Unity; all of them demonstrated interest in participating in the care of the newborn and expressed the need of information concerning to the health status of the newborn, the Intensive Care Unity environment and the hospital team. Several were the feelings expressed and the motives that indicated the needs of the mothers. PMID- 8715594 TI - [From experience to science: the evolution of nursing knowledge]. AB - The aim of this paper is to show the evolution of the nursing knowledge. Some relations with the nursing history are proposed, in order to indicate the ways the people and ideas of today in nursing was influenced by the past. The authors make a correlation between the development of the nursing as profession and of the nurse as professional. PMID- 8715595 TI - [Pattern of recovery of victims of cranio-encephalic injuries at 6 months and 1 year]. AB - A prospective and longitudinal study of head injury victims was conducted to identify the pattern of recovery 6 months and 1 year post-trauma. Patients with variety levels of injury and age between 12 and 60 years were observed. Functional limitation was analyzed using the eight-point Glasgow Outcome Scale (GOS). The majority of these victims had the following characteristics: age were < or = 35 years (68.6%); ISS > or = (51.9%) and Glasgow Coma Scale > or = 13 (72.2%) when they were examined by team of neurosurgeons at first time. Complete recovery or GOS 0 was achieved in 21.3% of the victims about 6 months after onset of injury and 38.6%, at the 1 year mark, Overall, the majority of victims made good recovery after 6 months post-injury, that is, GOS 0 or 1, and had returned to productivity in that period. The majority (65.0%) of victims reached their highest GOS outcome category by 6 months, although significant improvement was shown between 6 months and 1 year post-trauma. Similarly, return to productivity was higher 1 year after injury later than at 6 months. Findings indicated that GOS-GCS correlation was negative, and in some conditions GOS was correlated or associated with ISS and age. Return to work was not related to education level or job type. PMID- 8715596 TI - [Hospital infections in Brazil. Government activities for their control as an expression of socio-politics in the health field]. AB - The author studies the hospital infections evolution in Brasil and the demands for its control until its establishment by governmental actions. For a better understanding and analysis of the social actors' dispute process, their needs and interests and the relation with the governmental sector to the establishment of a certain state politics, a theoretical reference that enabled to elaborate the whole of social structure elements was searched. This theoretical reference should also take the filled of the relation between Society, State, Actors and Social Politics as its main question. In order to allude to these specific actions and its intervention range, it was necessary to make a distinction between the state politics modalities and the implementation responsibility. Based on empirical material, it was possible to trace these demands, conforming on technological-assistancial models through the working process indicators. Other empirical categories levels was demanded to approach to the theoretical reference: specific conjectures, welfare medical assistance, the clinical intervention model and the quality movement. PMID- 8715597 TI - [Forty years of history. Two or three reference points]. PMID- 8715598 TI - [Clothing symbols and professional identity]. PMID- 8715599 TI - [A new journal ... why? What does it tell about the professional identity of nurses?]. PMID- 8715600 TI - [A prospective glance at nursing care]. PMID- 8715601 TI - [Nursing diagnosis. It has come a long way]. PMID- 8715602 TI - [Urinary incontinence. Evolution of care from the dark period to nursing]. PMID- 8715604 TI - [Recognition of oncologic nurses]. PMID- 8715603 TI - [From care to education. The teaching nurse]. PMID- 8715605 TI - [Stoma therapy in France. Twenty years of evolution]. PMID- 8715606 TI - [The hospital hygiene nurse. Yesterday, today and tomorrow]. PMID- 8715607 TI - [From the neolithic to the 20th century. From the birth of agriculture to therapeutic nutrition]. PMID- 8715608 TI - [Stakes in education. Continuing education: why?]. PMID- 8715609 TI - [1955-2035]. PMID- 8715610 TI - [Nursing activities in type 1 diabetes mellitus]. PMID- 8715611 TI - [Child-oriented nutrition in diabetes mellitus]. PMID- 8715612 TI - [Requirements for ambulatory surgery from the viewpoint of the pediatric surgeon- experiences with ambulatory surgery in children]. PMID- 8715614 TI - [The scope of mentors' work at the Wurzburg University Hospital--education, tasks and activities]. PMID- 8715613 TI - [Experiences with ambulatory surgery in children--nursing care]. PMID- 8715615 TI - [Full-time work of the mentor: its sunny and its shady sides]. PMID- 8715616 TI - [A new era of acellular pertussis vaccines]. PMID- 8715617 TI - [Acute and rehabilitative care of children with prenatal infections]. PMID- 8715618 TI - [Confidentiality--the basis of trust in the care of the sick. II]. PMID- 8715619 TI - [Results of a basic questioning of pediatric hospitals, pediatric departments, pediatric-surgical hospitals and pediatric-surgical departments--the nursing part of the report]. PMID- 8715620 TI - Surviving the ax. Developing an education allocation system to verify productivity and resource consumption. AB - Workloads of nursing educators and nursing education departments as a whole have not always been captured accurately in the past, making it difficult to monitor and communicate resource consumption. Recognizing the need to determine an accurate Relative Value Unit in fiscally reliable terms, a Nursing Education Allocation System was developed to verify productivity and resource consumption. Standard values for each workload were established after a 6-month time study and entered into an automated allocation system. Productivity standards were then derived, making it possible to standardize measurement of workload from year to year and have good decision support data when determining staffing models in response to forecasted educational priorities. PMID- 8715621 TI - The retention of advanced cardiac life support knowledge among registered nurses. AB - In an investigation into the retention of advanced cardiac life support (ACLS) knowledge over time, the authors found that ACLS scores significantly decreased for a subgroup of a sample of 40 RNs employed in critical care areas in the first year after certification. Several variables that influenced scores were identified. Recommendations for inservice educators include reconstructing the ways in which ACLS courses are taught, conducting mock mega code scenarios every 6 months, constructing mega code scenarios that reflect the reality of practice, and routinely conducting refresher courses based on problems identified in a particular group. PMID- 8715622 TI - Preceptors' needs for faculty support. AB - When practicing nurses accept responsibility for preceptoring nursing students, they use and develop the teaching skills that are part of their professional practice. However, they need both support in the role and mentoring to develop their skills. Inservice educators and experienced preceptors often provide that support and education. Based on a qualitative study of preceptoring nurses, the author explores preceptor needs for support, the type of support required, and faculty roles in the provision of this support. PMID- 8715623 TI - The relation of work stress, hardiness, and burnout among full-time hospital staff nurses. AB - In this descriptive study, the author examined the relation of work stress, hardiness, and burnout among 113 full-time hospital staff nurses. It was hypothesized that among hospital staff nurses: 1) the greater personality hardiness, the less job stress; and 2) the greater personality hardiness, the less burnout. Instruments used were Personal Views Survey, Nursing Stress Scale, and Tedium Burnout Scale. Promoting hardiness through nursing staff development programs may help manage stress and reduce burnout in the healthcare setting. PMID- 8715624 TI - Head nurse to nurse manager. Easing the transition. AB - At a large, teaching healthcare facility, a nursing leadership task force designed an expanded role for head nurses, broadening their organizational responsibilities to enhance the quality of care, quality of work life, and operations. Acknowledging the limited educational background in management skills, the nurses needed additional instruction to perform competently and efficiently. An educational program was developed to facilitate the transition from head nurse to nurse manager. PMID- 8715626 TI - Continuing health education via interactive television: a pilot project. PMID- 8715625 TI - Certification. Hospital versus national standards. AB - "Certification" and "credential" are frequently used terms in nursing. How these terms are used and what they mean to nursing practice is the focus of this article. Definitions, history, and application of these concepts are explored. Recommendations are made that a hospital not assume responsibility for the granting of certification or credentials to nursing staff members. Nurses and nursing administrators are urged to support nationally recognized certification programs. PMID- 8715627 TI - Telling your story. PMID- 8715628 TI - Right to die issue raises complicated ethical questions. PMID- 8715629 TI - ONS drafts position statement on the use of placebos for pain management in patients with cancer. PMID- 8715630 TI - Professional liability: to have? Or not to have? Part 2: Qualified immunity. PMID- 8715631 TI - [Revisiting our values. A critical look at the ordinary life]. PMID- 8715632 TI - [Hospital: values of a teaching enterprise]. PMID- 8715634 TI - [Values in professional and social life]. PMID- 8715633 TI - [More care for less expense?]. PMID- 8715635 TI - [What did I do to my values?]. PMID- 8715636 TI - [The uniform as a nursing value]. PMID- 8715637 TI - [New perspectives for the profession of nurses' aides. Evolution towards new values. Mutations?]. PMID- 8715638 TI - [Taking care of oneself ... in order to take care of others]. PMID- 8715640 TI - [Nursing diagnosis. On what really depends its integration into nursing practice?]. PMID- 8715639 TI - [University diploma in health ethics]. PMID- 8715641 TI - [From nursing delegation to collaboration. New study program for nurses' aides]. PMID- 8715642 TI - [Letter to a school nurse]. PMID- 8715643 TI - [Pedagogy and pedagogue]. PMID- 8715644 TI - [The place of taboos in nursing practice: results]. PMID- 8715645 TI - Parents' values: a missing link in the neonatal intensive care equation. AB - Neonatal intensive care has become a widely accepted component of our health care system. Its acceptance and growth are related to increased technology and specialization. However, little consideration has been given to the process of making decisions about the use of neonatal intensive care from the perspective of the patient's family. This article reviews the existing literature related to parents' opinions and perceptions of care in the NICU and proposes a framework for the exploration of the value systems of parents of infants requiring neonatal intensive care. PMID- 8715646 TI - Percutaneous central venous catheter-related sepsis in the neonate: an analysis of the literature from 1990 to 1994. AB - Percutaneous central venous catheters (PCVCs) have been used in neonates since the 1970s. During the 1980s, they were introduced in many NICUs. Most studies published to date employ a descriptive methodology. There are very few randomized clinical trials with PCVCs in neonates, and no integrated literature reviews have been published to date. Furthermore, infection in neonates with PCVCs has become a major concern. This integrated literature review was conducted to delineate the scope of the problem of sepsis in neonates with central venous catheters and to identify current research directed at methods to reduce catheter-related sepsis (CRS) in high-risk neonates. Twenty-five references were found and analyzed for this review. The definitions used to identify CRS in neonates varied greatly among studies. The method of calculating the CRS rate varied as well. The CRS rates ranged from 0 to 29 percent or from 0 to 15.3 infections per 1,000 catheter days. Strategies employed to reduce CRS rates are presented along with recommendations for future research. PMID- 8715647 TI - Liquid ventilation: a comprehensive overview. AB - Despite advances in neonatology, some infants do not respond to current pharmacologic and ventilatory techniques. Others suffer chronic lung disease, require prolonged ventilatory support, and experience significant morbidity during infancy due to the elevated inflation pressures used to treat their respiratory problems. Over the past 30 years, results of studies in premature animals as well as clinical trials have demonstrated that ventilation with oxygenated perfluorochemical (PFC) fluids provides effective gas exchange and improved lung mechanics. PFC fluids are biologically inert, have a high gas solubility and a low surface tension, and are nonbiotransformable. With liquid ventilation, alveolar pressures are low because the high surface tension of the gas-lung interface in eliminated. Potential neonatal applications include surfactant deficiency, persistent pulmonary hypertension, meconium aspiration, diaphragmatic hernia, pneumonia, and a vehicle for drug delivery. In order to develop a nursing care plan for the liquid-ventilated infant, nurses need knowledge of the physiologic changes involved in liquid ventilation, as well as its mechanics. PMID- 8715648 TI - Sources of support reported by mothers and fathers of infants hospitalized in a neonatal intensive care unit. AB - Over the past two decades, awareness of the importance of social support for individuals faced with a major life transition or a stressful event has increased. The purpose of this study was to identify the perceptions of mothers and fathers of critically ill infants about the helpfulness of support provided to them by family, health care professionals, and other when their infants were in a neonatal intensive care unit (NICU). Social support in this study is conceptualized as the interpersonal transactions that parents of preterm infants perceive as helpful in reducing their stress and coping with their child's illness. Data were collected using the Illness Support Scale, on which subjects rated the helpfulness of support from individuals across their network. Findings indicate that these parents experienced a moderately high level of support and perceived NICU nurses as very helpful. Nurses need to continue to develop their role in helping families by identifying specific supportive interventions. PMID- 8715649 TI - Securing ET tubes. PMID- 8715650 TI - What is Healthy People 2000? PMID- 8715652 TI - Neonatal intensive care in the Ukraine. PMID- 8715651 TI - Developmental care: first things first. PMID- 8715654 TI - Neonatal immune thrombocytopenia: isoimmune or autoimmune? PMID- 8715656 TI - Research utilization: the NICU environment. PMID- 8715655 TI - Morphine sulfate. PMID- 8715657 TI - Loving Stephanie. PMID- 8715658 TI - [Isolation room and intensive care room]. PMID- 8715659 TI - [Working out a research project]. PMID- 8715660 TI - [The regulatory problem]. PMID- 8715661 TI - [A meaning to be clarified]. PMID- 8715662 TI - [Practical modalities]. PMID- 8715664 TI - [A clinical position]. PMID- 8715665 TI - [Use of isolation rooms]. PMID- 8715663 TI - [One isolation may bring on another]. PMID- 8715666 TI - [Psychiatry without writing]. PMID- 8715667 TI - [A threatened man]. PMID- 8715668 TI - [Reflections on the "creature nurse"]. PMID- 8715669 TI - What you can tell women about hormone replacement therapy. PMID- 8715670 TI - [The interaction between language and profession]. PMID- 8715671 TI - [Professional lingo in nursing]. PMID- 8715672 TI - [Linguistic competence and emancipation]. PMID- 8715673 TI - [A message from expert to expert: nursing documentation]. PMID- 8715674 TI - [Career planning within the nursing service]. PMID- 8715675 TI - [Stopping treatment in patients with incurable diseases--mercy killing or homicide?]. PMID- 8715676 TI - [Nursing measures for the prevention of constipation]. PMID- 8715677 TI - [Funeral rites in memory of the elderly]. AB - This a phenomenologial study rites in the memory of elderly people, originated from the discomfort lived by the authors in their professional life dealing with death and dying. Verbal information from elderly people was collected with the objective of recovery and decoding mortuary rites. Nine themes originated from these informations: feelings and meanings in relation to death, the time of death, the annunciation death, the body's preparations, the watcher, the funeral procession, the grave, the return to home, the remembered death. The results gave the authors opportunity to understand better the attitudes of health professionals in caring for patients and their families in this existential experience of to-be-for-death. The death rationalized by scientific knowledge and nonpersonal technological care hides new rites, transmuted by new representations which the society built. PMID- 8715678 TI - [Psychiatric nursing practice in Ceara and its relationship with socio-historical contexts]. AB - The study focuses on the practice of nursing assistance in psychiatric in Ceara. The data was collected by interviews and documents regarding history of nursing and of the assistance psychiatric in Ceara. On conclusion, the nursing practice is essentially supervision to the nursing team in delivering psychiatric medicine. The nursing practice has specific data to be considered on its study as whole: the relationship between psychiatric assistance and society. PMID- 8715679 TI - [Perspective evaluation of the post-graduate courses of the Nursing Department at UFRN (1980-1992)]. AB - The purpose of the study was to evaluate the "lato sensu" graduate programs of study of the Department of Nursing in the UFRN, and to know the perspectives of its students and faculty. A total of 67 questionnaires were applied to 14 professors and 53 ex-students. The sources of data were: a survey of the scientific knowledge produced by the faculty; specialization program final reports; partial 1989-1991 program evaluations; and, the final report of the first Graduate Program Evaluation Seminar (1992). Eventhough the results of the evaluation were positive, some aspects of the program were found to need further analysis so that future graduate studies might be improved. These aspects were: revision of the curriculum content, improvement of the space environment, and improvement of the bibliographic resources. PMID- 8715681 TI - [Preoperative orientation of cataract patients and indications for ambulatory surgery--report of experiences]. AB - The study describes the nursing care of patients with cataract before they are submitted to ambulatory surgery. The procedure involves a systematic orientation program which is being developed by the authors, in groups with 15 patients each. The objectives are: to provide technical advice on cataract, to instruct the patients as to nursing orientation care, to enable a nursing evaluation of these patients and to help reducing the number of absent patients and cancelled surgeries. The authors analyze the patients' concerns with their ocular problem, their surgery and post-operative care. The limitations were pointed, out, concluding that de program is relevant for the patient with a more effective participation and adhesion to the treatment. PMID- 8715680 TI - [Teacher-student communication across the Tizadora pedagogical problem]. AB - Report of experience in the use of a social-cultural approach during the development of a work done by graduate students of Ribeirao Preto School of Nursing-USP. As a form of reference for the didactic strategy, it was used the scheme of arch proposed by Charles Maguerez and presented by BORDENAVE, in an attempt to systematize the communication teacher-student talking about "Aspects related to the Undergraduate Teaching Problems". PMID- 8715682 TI - [Self-care in treatment following the Ilizarov method--a case study]. AB - The authors performed a case study using Dorothea Orem's self-care model as a framework in the treatment by llizarov's method, administering care and specific orientations for recovery and prevention of further disorders. The operational system of the study consisted of procedures related to demands of universal and therapeutic care during hospitalization. Demands were detected in the pre operative and care developed in the postoperative phase, providing the patient with the knowledge and achievement of the necessary procedures to be followed at home. PMID- 8715683 TI - [Education and social exclusion of women: a gender problem?]. AB - Using gender as an analytic category, this paper shows the way the social relations between men and women operate historically and how the female values give place to the male ones. In such context, the educational process has a special importance as one of the determinants of the woman social inclusion/exclusion. PMID- 8715684 TI - [Ostomy: limitless rehabilitation? The nurse's point of view]. AB - Knowing about the concept of rehabilitation and the semantic and philosophic meaning of the word limit (frontier), the author gives five basic propositions to justify the process of caring for rehabilitation of the ostomy patient. Concluding, the author states that this process has as limit its starting point. Therefore, it has to begin at preoperative stage. On the other hand, the limit as a point of arrival will established by the ostomy patient's motivation and his/her capacity of being involved in the self care activities and social participation. The author also declares that when rehabilitation is reached as a result of rehabilitation assistance process, it will be part of the dynamics of interpersonal relationships focusing on a new life style or a new social reality and, finally, as part of the process of living the ostomy person. PMID- 8715685 TI - [Support system for patient care planning and prescription for nursing care]. AB - The use of information technology is reduced in health area. Its introduction as a management tool and decision support can change the actual kinds of planning within health organizations. The purpose of this work is to develop a system in order to enable and process the nursing prescription. The system named DSSPPNC Decision Support System in Planning and Prescription of Nursing Care-is a computational tool that enables the information access under the form of Nursing Assistance Protocols. The results of the present work will involve sensible changes in the nursing assistential practice, with repercussion in the type of assistance management and to care the individual hospitalized, making opportune and approaching the nurse to the patient, through raising of problems, aiming care planning and accomplishment, results valuation and supervision and control of the staff engaged to in assistance. PMID- 8715686 TI - [Modality of support for research in the field of nursing]. AB - This paper deals with the organization of the Operational Research Committee implemented at the University Hospital of the University of Sao Paulo in 1993 with the objective of giving logistic support to nurses in the development of their scientific work. It shows the strategies adopted to encouage research among nurses to reach the success of the proposal. PMID- 8715687 TI - [Ethics in nursing]. AB - Synopsis about the evolution of moral principles adopted by brazilian nursing since 1923 to nowadays. It indicates a metaphysical, abstract and spiritual tendencies in the codes of 1958 and 1975, and a change in the code of 1993, which searches to observe valuables of historic form and based on material conditions of existence. PMID- 8715688 TI - [Emotional support offered by the nursing team to children with cancer and their family]. AB - The purpose of this paper is to unveal of the emotional support provided to children with cancer and their family by the nursing staff. By this, a qualitative methodology was applied, making possible a comprehensive analysis of the statements of the nursing staff that living this situation. The convergence of these speeches were analyzed, enabling the identification of some significant units that can give subsidies to guide the emotional support provided by the nursing team. PMID- 8715689 TI - The sacred cows of asepsis. PMID- 8715690 TI - Bowel technique in the O.R. Is it really necessary? AB - Existing inconsistencies in the practice of bowel technique prompted a study to validate this operating room procedure. Minimal reference to intra-operative bowel technique was found in the literature. Needle drivers and tissue forceps utilized to anastomose large bowel were cultured, and the results were analysed. Needle drivers and tissue forceps used to anastomose small bowel were also cultured and used as a control group. Only elective bowel surgery cases were included in the study. The study demonstrated that there was consistent contamination to those instruments used for bowel anastomosis, with significantly greater contamination for large bowel resections. These results indicate that isolating those instruments and equipment which come into direct contact with the bowel lumen is a perioperative practice which will reduce the surgical patient's risk of postoperative wound infection. PMID- 8715691 TI - Planning for the profession: recruitment, retention, research, resources. PMID- 8715692 TI - Measuring quality: evaluating personnel competencies. AB - Historically, quality and performance evaluation have been considered a management responsibility and have been completed retrospectively. In this article, I am proposing a radical shift. Specifically, I am encouraging the elimination of performance appraisals and, in their place, the introduction of continuous progress reviews and competency assessments which are initiated by each individual, not by management. Quality care will continuously improve in direct relationship to improved personnel competence which can be measured by using established quality indicators and expected client outcomes. PMID- 8715693 TI - Eye glasses and dentures in the O.R.: the choice is yours! AB - Historically, patients have been required to remove their dentures and eye glasses on the pre-surgical ward before being taken to the Operating Room (OR) on the day of surgery. This descriptive study examined the choices made by patients pre-operatively regarding dentures and/or glasses, the reasons given for the choice, and the relative satisfaction with the choice post-operatively. Of the 213 patients enrolled in this study, 66% were female with an average age of 57 years. Only patients wearing eye glasses or dentures were included. Sixty-nine percent of the patients wearing glasses and 85% wearing dentures chose to wear them to the OR. When asked post-operatively about their satisfaction with their choice, 95% indicated that they were very satisfied with their choice and that it was the best decision they could possibly have made. No glasses or dentures were broken or lost during the study. PMID- 8715694 TI - The future of long-term-care case management. PMID- 8715695 TI - When case management isn't enough: a model of paraprofessional advocacy for drug- and alcohol-abusing mothers. AB - The Seattle Birth to 3 Project is a paraprofessional advocacy model of enhanced case management designed to effectively intervene with high-risk, drug-dependent mothers. Postpartum women were enrolled based on their heavy use of drugs or alcohol during pregnancy and lack of connection to community services, including prenatal care. Each participant was assigned a paraprofessional advocate who worked intensively on a one-to-one basis with her and her family for 3 years postpartum. Six components of the model advocacy program are identified: establishing the relationship; identifying client goals; establishing linkages with service providers; using written agreements; role modeling and teaching of basic skills; and evaluating the outcome. Results for 51 women assessed after 1 year indicate significant areas of improvement including increased involvement with drug/alcohol treatment agencies, decreased drug use, increased use of birth control, and increased involvement with supportive and skill-building groups such as parenting classes. PMID- 8715696 TI - Ohio's options for elders initiative: cutting corners or the cutting edge? AB - This article presents results from the evaluation of Ohio's community-based long term-care demonstration project. The initiative, named "Options for Elders," was an attempt to develop a more comprehensive and rational service delivery system for disabled older people. The demonstration involved a single point of entry to long-term-care services, a telephone screen, varying levels of care management, and funding for services. The needs of clients were initially assessed by telephone, and clients were assigned to levels of care management and services based on the concept of medical triage. By placing additional responsibility on a telephone screener, the Options approach attempted to reduce the amount of in home care management provided. The model is a unique and controversial attempt to develop a more cost-effective system of care management. PMID- 8715697 TI - Downsizing and diversion: strategies to reduce Medicaid long-term care expenditures. AB - In their efforts to control escalating Medicaid costs, some states have focused on shifting their long-term care system from predominantly institutional services to home- and community-based services. Two strategies states have employed that may be of interest to case managers are: (a) downsizing the overall supply of nursing home beds and (b) diverting consumers from nursing facilities before they are admitted. Both strategies of downsizing and diversion can impact case managers in that the array of services from which they can choose and the process by which they plan their clients' care can be affected. This article describes some of the approaches that states have used to decrease the use of institutional services including bed moratoria, bed buy-back, and pre-admission screening; the problems that they have encountered; and the role of case managers in assisting states to ensure access to services while controlling costs. The article summarizes some of the findings presented in Reducing the Cost of Institutional Care: Downsizing, Diversion, Closing and Conversion of Nursing Homes, which was developed for a meeting sponsored by the University of Minnesota/National Academy for State Health Policy National LTC Resource Center, entitled "The Cost of Institutional Care: How to Get Around the Roadblock to Medicaid Reform," funded by the U.S. Administration on Aging. PMID- 8715698 TI - A method to determine case manager caseloads in long-term care. AB - Case management has come to the forefront of home- and community-based long-term care (LTC) as an attempt to reduce the costs of LTC and the fragmentation of the home- and community-based LTC system. The cost of case management itself, however, must also receive attention. A key variable contributing to the cost of a case management program is the size of the case managers' caseloads; the higher the caseloads, the lower the costs, all other things being equal. However, caseloads that are too high can reduce the quality of care. Thus, a program's recommended caseload is an important factor. This article presents a method for determining a recommended caseload for a home- and community-based LTC program, using a case study. This method can be adapted for use by programs that have a different caseload history from that of the program in the case study. PMID- 8715699 TI - Care planning in home care: an exploratory study. AB - Care Planning is one of the most important responsibilities of the case manager in home care. Research on how care plan decisions are made is scarce in the case management literature. The purpose of this exploratory study was to determine the factors that influence the decision-making process of case managers within the Edmonton Home Care Program (EHCP) when developing care plans. A volunteer sample of six EHCP case managers from three different disciplines was selected: two registered nurses (BScN), two occupational therapists (BScoT), and two social workers (BSW). With the use of fictional case studies, these professionals were asked to construct care plans. Individual semi-structured interviews and a group session permitted in-depth exploration of their decision-making process. Findings suggest that client and/or caregiver characteristics appear to be the most influential in determining care plans. It was also found that case managers are not comfortable with fiscal accountability. PMID- 8715700 TI - Nurse entrepreneurs--is the public willing to buy their services? PMID- 8715701 TI - Ethical decision-making for nurses in Alberta: guidelines and recommendations. Alberta Association of Registered Nurses. PMID- 8715702 TI - Working women and health problems. PMID- 8715703 TI - Report on the Fourth World Conference on Women, '95. PMID- 8715704 TI - Nurses make the difference in health care. PMID- 8715705 TI - Finding the multimedia resources you need. PMID- 8715706 TI - Strategic planning: outmoded concept or current imperative? PMID- 8715707 TI - A shared nursing vision: essential in a learning organization. PMID- 8715708 TI - Reinvent your staff development department. PMID- 8715709 TI - Measuring education: tracking for change and business results. PMID- 8715710 TI - Rebecca reminds me of the essence of oncology nursing. PMID- 8715711 TI - We need to understand "change + human beings = transitions". PMID- 8715712 TI - If I can do it, so can you. PMID- 8715713 TI - [Nursing science in the service of the community]. PMID- 8715714 TI - [Choosing nursing]. PMID- 8715715 TI - [Postgraduate nursing studies/third millennium]. PMID- 8715716 TI - [Team work: a basic instrument and a challenge to nursing]. AB - Following the literature review, it was verified in previous study, the way the nurses working governmental hospitalization unit, characterize and experience team work. PMID- 8715717 TI - [Closed endotracheal suction system for the prevention of hypoxemia]. AB - This paper shows a revision of the studies about indications efficacy and dangerous effects from the use of the closed tracheal system suctioning. In spite of the difficulties in the generalization of the results of the analysed works it was possible to conclude that the "trach-care" has its comproved efficiency in the maintenance of PaO2 in most intubated and ventilated patients. PMID- 8715718 TI - [Mothers' clubs: a place for women's health care]. AB - The article relates an investigation realized next to integrant of Mother's Club of the Sanitary District of Perus, in the Municipal of Sao Paulo. It has how aims to know the its organizational shape and the relation between locals Municipals health institutions and with other women movements. With this to intended to know the concepts about health disease from their members to verify the possibilities of intervention in coletive health. The results reveal in the conceptions of health-disease of their integrants. Observe, however, spaces to intervention in the feeling to mplify the breast-plate of the women about of the their social identity and of the woman's health. PMID- 8715719 TI - [Nursing care for women during climacteric]. AB - Climateric is a transitory stage in women's life, which causes remarkable alterations. This article brings into focus how important the nurse performance is, giving protection, promotion and recuperation for women's health. The authors suggest some recommendations during nursing care. PMID- 8715720 TI - [Aspects of the social reintegration of stoma patients]. AB - The social rehabilitation represents a challenge to ostomy patient and a concern for health professional team. This study intends to verify the difficulties related to returning to the daily domestic, leisure, job and sexual activities showed by 45 ostomizeds after hospital discharge in different periods of late post operative. The data was obtained from patients assisted by two Outpatient Services of Sao Paulo city, through the interviews. The patients were selected by previous established criteria agreement and then subdivided in three groups as the post operative period. It's possible to verify that the majority of the clients didn't returned totally or returned only partially to the daily activities like before the disease or surgery, specially in the sexual area. They justified these difficulties by physical problems, insecurity and the use of inadequate appliances. PMID- 8715721 TI - [Nurses' uniforms as seen by patients, nurses and nursing faculty]. AB - The exact role of the way people dress up on the nonverbal communication is still unknown. Nevertheless, we know that it influences the interpersonal answers and, in some situations, they are the main determinants of those answers. The objective of this study was to determinate differences between the perception os the nurse uniform through the answers of 100 patients, 30 nurses and 15 nursing school faculty. The data were collected by showing nine photographs of hospitals of the city of Sao Paulo (Brazil). White trousers and blouse was the favorite one among patients, nurses and nursing school faculty regarding personal care. White skirt above knees, white blouse and blazer was the most rejected one by three groups. PMID- 8715722 TI - [The nurse's role in the recovery room--the best assistance during the perioperative period]. AB - The article describes the nurse's role in the recovery room about assistance, educational, administrative and research areas. Those areas explored in our country to improve the assistance in perioperative period. PMID- 8715723 TI - [Living with diabetes: an ethnographic study]. AB - The purpose of this study was to learn the experience of diabetic clients who live health-disease phenomena in an outpatient service of endocrinology in a school hospital. The objective of the research was to obtain a comprehensive view of their way of thinking their practices and their feelings lived during treatment. I adopted SPRADLEY's ethnographic methodology which includes: participant observation and ethnographic interview which are carried out simultaneously. Symbolic interactionism was also a strategy to clarify the themes, the validation of domains pointed out to the central theme: "The disease came to stay". The diseases metaphor emerged from this cultural universe as dangerous, impossible that destroys and stays. The disease is lived as a illness that provokes different feelings like sadness, panic, pain and death. The emic knowledge obtained from this ethnographic research gave to nursing practice important episthemologic and onthologic bases. PMID- 8715724 TI - [Nursing graduation: the new curriculum]. AB - The research shows the up date curriculum of nursing Graduation and the basis of this proposition. PMID- 8715725 TI - [Evaluation of the quality of public health care for children from the user's viewpoint]. AB - This paper presents results from research that used a qualitative approach to identify evaluation criteria from an user viewpoint, and tries also to evaluate the work of the Whole Care for Children's Health Program (WCCHP), which directs actions in the children care services at the basic level. Dates were obtained th partly structured interviewing at home with ten adults in charge of children registered in October 1992 at four units of the Barra-Rio Vermelho District. Through a simultaneous process of gathering and analyzing dates, using a content analysis technique it was possible to classify the results in five groups: "motivations" to look for the service; having expectations; "choosing" a health service; evaluating the service used; and suggesting improvements. The evaluation criteria found are closely related to: access; interpersonal relation; results; and continuing care, which possess different value degrees according to the motives for service need. The results of the evaluation show that the model of clinical assistance and the form organization of the services do not fulfill the objectives of PAISC, breeding access difficulties, and poor results on account of the lack of material resources, coordination and care continuity. PMID- 8715726 TI - [Study of the AIDS epidemic in the city of St. Carlos]. AB - This study emerged in the face of the need to consolidate the information from the clinical HIV+ and AIDS cases recorded at the Ambulatorio Regional de Especialidades de Sao Carlos (Special Clinic). Its aim is to identify the population according to sex, age group, marital status, level of education, occupation and the variables related to the epidemic means of exposure, laboratory tests and disease status-as well as to provide the health service with information. The data from 204 patients, 147 males and 57 females were analysed; most people were between 20 and 30 year of age and single. For females, the predominant mean of exposure was heterosexual sex, and for males blood transmission (intravenous drug use) predominated. With this analysis it will be possible to follow the development of the epidemic in the region and to propose preventive measures, which are considered to be only option for control of the disease at present. PMID- 8715727 TI - [Surgical wound infection--a complication of mastectomy]. AB - The present study contains a retrospective analysis of thirty medical reports of patients that were submitted to mastectomy at a school hospital, and has the objective of knowing the frequency of surgical injury infection, relating it to predisposed factors to infection. The frequency founded was 40%, a percentage above the level pointed in the literature, suggesting that can be included in it cases of seroma, due the indicators utilized to define infection and the absence of laboratory exams registers in the consulted medical reports. PMID- 8715728 TI - [A play technique as a new approach to health education]. AB - This paper presents the premisses and results reached in the first phase of a community project of the College of Health Sciences of the University of Brasilia, whose purpose is to develop a new approach to health education based on the use of instruments and teaching techniques involving play. For this phase the objectives were: to set up a play room in the satellite city of Paranoa-Federal District; to make the teachers aware of the importance of play in the development of children; to improve their role in this kind of work with children; to develop new maintenance of the play center. The authors relate their experiences in the project in the period of a year and a half, pointing out certain factors which facilitated and inhibited the results, as well as explaining how they propose to use this space to develop a new approach to health education with a focus in health promotion. PMID- 8715729 TI - [Use of papain in visceral infections]. AB - This research was carried out at the University Hospital of the University of Sao Paulo and deals with the experiment of papain utilization for visceral irrigation in patients with severe infection. It was observed that seventy two hours after treatment, there was a considerable reduction of purulent secretion, and that the medium time of cicatrization of all lesions was thirty days. PMID- 8715730 TI - [Research in the past]. AB - The present study based on the literature, has the purpose of exploring research, it stages, and the types of potential problems that can interfere on its achievement. It also tries to bring some studies about Brazilian nursing history, pointing out some considerations, as well as the interrelationship of nursing and its history in the search for science. PMID- 8715731 TI - [Philosophy of nursing education. Various considerations]. AB - The study has objective to promote a reflection about the importance of philosophy for nurse's formation. For this purpose, we search of questions in the literature that could give foundations about the theme, focusing on the importance for scientific nursing research. PMID- 8715732 TI - [Research as an instrument in practice]. AB - This paper speaks about the utilization of research as an instrument in the nursing professional practice in the areas of assistance, administration, and teaching. It situates science as a process and emphasizes nursing responsibility in the production and utilization of the scientific knowledge. Also stands out factors that have same positive or negative influence in the scientific research, besides ethic aspects involved. PMID- 8715734 TI - [Reflections concerning the concept of daily work (according to Agnes Heller and Michel Maffesoli)]. AB - Synopsis about writings of Agnes Heller concerning the work category, with the purpose to point out parallel between her opinions and Michel Maffesoli's. There is evidence that in both are manifests the analysis of daily life, historic space of individual and social development. Heller works this category-daily-into an approaching "constructivist - dialectic", where the subject of historic is visualized in its temporality (past, present and future) into the process of individual and collective changes. The approaching of Maffesoli has a nucleus "phenomenological Comprehensive", where the historic subject is persued in the dimension of present ("usual life/banal life). PMID- 8715733 TI - [Osteoporosis: a silent epidemic which will turn public]. AB - This paper presents a bibliographic review about osteoporosis, point out the concept, physiopathology, types and risk factors. It emphasizes how the health team must develop the assistance according the different types of osteoporosis, mainly the preventive measures that can retard or even stop this disease, like: the supplementation of calcium and vitamin D in the diet if necessary, the hormonal therapy and physical activity. PMID- 8715735 TI - [The students' page. Notes about patient records and the production and reproduction of knowledge. Written and oral presentations]. AB - This paper relates a description about nursing notes in pactients promptuaries in relation to the distribution/reproduction/production process of knowledge in nursing. It was developed on an Intensive Care Unity from a Hospital in the teaching/assistance Sanitary District Barra/Rio Vermelho (Salvador-Bahia). The basic premise is that the nursing personal work superficial and sporadic notes just related to patients. Distribution/ reproduction/production process, that happen in the nursing work process isn't considered by it devaluation or just because, for nursing agents, only information about technical procedures related to patient care, expresses the nursing knowledge and so, must be registered. In order to reach the objective, 30% of the promptuaries of patients discharge from the ICU were studied and the nursing team was interviewed, during the months november, december 1994. The analysis indicates that the way how the nursing notes are been made, results in hard communication among agents of health area and contributes to disqualify the nursing assistance that is given to patients, besides to limit the advance of the distribution/reproduction/production of knowledge in nursing. PMID- 8715736 TI - [Facial expressions and human emotions. Bibliographic survey]. AB - The present work intends to divulge some Nursing and Psychology' publications about human facies and emotions. Intends do help the health professional thinking about the facial expressions of emotion in the interpersonal relationships, showing the researches more cited at specific bibliography, first regarding to children and to adults. PMID- 8715738 TI - Improving the standards of care delivery in mental health. PMID- 8715737 TI - [The "crazy" person, the expert revision: the humanization of psychiatry]. PMID- 8715739 TI - Clinical guidelines: risk management and legal issues. PMID- 8715740 TI - Nursing management of patients receiving gastrostomy feeding. AB - A feeding gastrostomy should be considered for patients who require enteral tube feeding for more than 2 weeks. This article describes three types of gastrostomy: percutaneous endoscopic gastrostomy; replacement/surgically placed gastrostomy; and skin level/button gastrostomy. The choice of tube and method of insertion are governed by the patient's clinical condition and lifestyle. General guidelines on the decision to initiate gastrostomy feeding are given, together with indications, insertion and nursing management for each tube. PMID- 8715741 TI - Treatment of venous leg ulcers: 2. AB - This article, the second in a two-part series, discusses the treatment options available for venous leg ulceration, and their implications for the nursing profession. Current views on the use of compression therapy, drug therapy, growth factors and surgery are examined. Successful management of patients with venous leg ulcers depends on thorough assessment, effective treatment and prevention of recurrence. Many health-care professionals lack the necessary knowledge, understanding and training to treat such patients effectively. Improvements in the care of venous leg ulcers will lead to significant reductions in treatment costs. PMID- 8715742 TI - Self-administration of drugs on an oncology ward. AB - This article focuses on the development of a programme of self-administration of medicines within an oncology setting. Previous studies which have developed this concept demonstrate that such a programme has a positive impact on a patient's knowledge of his/her illness and treatment. Compliance with drug regimens increases and patients are given back control over one aspect of their lives while in hospital. The development of an individualized drug administration programme within an oncology unit and how it will be evaluated in the future are described. PMID- 8715743 TI - The role and criteria of an advanced nurse practitioner. PMID- 8715744 TI - The research role of the advanced nurse practitioner. AB - The research role of the advanced nurse practitioner (ANP) is poorly developed in the literature. Clarification of the role is essential if individual nurses are not to be burdened with unrealistic expectations. Organizations need to prepare themselves for research activities and must not expect the ANP to address all the issues involved alone. PMID- 8715745 TI - Advanced practice within learning disability nursing. AB - This article describes the application of advanced nursing practice within the field of learning disabilities. It discusses the difference between advanced practice and specialist practice and proposes that managers' perceptions of the role of advanced nurse practitioners (ANPs) within the management of developing services will need to change if such practitioners are to be effective. Six defined areas are explored to show how advanced nursing practice within learning disabilities may be developed to allow ANPs to assert their rightful position and influence. PMID- 8715746 TI - Role of the nurse teacher in advanced nursing practice. AB - Education and experience are the foundations on which practitioners should base their clinical judgment and broaden the scope of their professional practice. PREP has secured a professional market for nurse education, the viability of which depends on closer collaboration between nursing education and nursing practice. Nurse teachers who are committed to practice are in a strong position to assist the development of practitioners, and work towards research-based practice. This article examines the role of the nurse teacher in the domain of advanced nursing practice. PMID- 8715747 TI - Study of moving and handling practices on two medical wards. AB - This small exploratory pilot study was undertaken to explore the moving and handling practices of nurses working on two medical wards and to determine nurses' perceptions of factors that might influence those practices. Data were collected by means of non-participant observation and semi-structured interviews. The results suggest that risk assessment of the task, load, environment and individual capabilities, carried out in the clinical area, was often incomplete. Factors influencing moving and handling practice included insufficient equipment, lack of space, unsuitable uniforms, and negative attitudes towards changing practice. In view of changing practices, nurses need to be aware of the factors that promote or hinder moving and handling practice if they are to address these issues. PMID- 8715748 TI - Are nurse administrators losing touch with nursing? PMID- 8715749 TI - Pressure sore prevention: using and evaluating risk assessment tools. AB - The Department of Health issued clinical practice guidelines in June 1995 concerning the prevention and treatment of pressure sores (United Leeds Teaching Hospital NHS Trust, 1995). The guidelines were produced through a combination of review of the research evidence, debate in expert panels and wide consultation (Hitch, 1995). This article reviews the section of the guidelines concerning risk assessment. It looks at the research evidence evaluating risk assessment tools, noting problems which exist in interpreting the evidence, and the conclusions that can be reasonably drawn from it. It also discusses the idea of developing and evaluating risk assessment protocols based on routinely used risk assessment tools to ensure that the information acquired through such assessments is used appropriately to allocate effective preventive interventions. PMID- 8715750 TI - Meal skipping patterns and nutrient intakes of rural southern elderly. AB - Dietary practices, a 24-hr food recall, self-reported health status, perceived well-being, and socioeconomic data were collected from 2890 rural elders selected from 11 southern states by a stratified randomization method based on income and population. Overall nutrient intakes were low, especially energy and calcium, and 16.3% of respondents consumed fewer than three meals daily. Though meal skippers snacked more frequently, their nutrient intakes were significantly lower than those of three-meal eaters. Meal skippers were more likely to be smokers, younger elders, females, those who ate alone and those with less education, higher BMI, and lower self-rated quality of well-being, especially economic and social well being. PMID- 8715751 TI - Food management for the aging population. AB - The older population is becoming more important to our society everyday. These individuals are being studied for their past, present, and potential impact on markets and marketing. Evaluated as a user of products or services in the marketplace or an employee or volunteer within the marketing system, this segment is gaining a visibility and importance. An interview was conducted with five Nutrition Project Directors to obtain an overview of Federally Funded Nutrition Programs for the Elderly. The areas which were highlighted were service delivery, site activities, management styles, barriers to service, clientele composition, food planning and preparation, staffing, USDA funding, coordination, marketing, transportation, and volunteerism. The Second Quarter Service Provider Output Reports for 1991, which are compiled by the Nutrition Projects and submitted to the Area Agency on Aging, were utilized to obtain client profile information (Reports, 1991). The analysis sought to compare the programs offered in the five counties on a number of factors which could be quantified. It was hoped that by looking at the numerical ratios, and depicting them graphically, any trends or unique characteristics of the programs could be identified. In that the percentage of Florida's present elder population (17%) far exceeds the national average (12%) these findings could be utilized by nutrition programs outside of Florida to plan for future funds. Analysis of quantitative information on the five programs yielded information on cost comparisons and on services. PMID- 8715752 TI - The resident dining room--a CQI project. AB - The resident dining room was noisy, food was served cold and late, food containers were hard to open, and food preferences were not respected. Verbal and physical confrontation was common, there were no specified meal hours, and care givers were not available to assist residents during meals. A multidisciplinary Continuous Quality Improvement (CQI) committee was convened to look at the situation. Brainstorming and prioritizing resulted in a list of items to be addressed. A plan was developed and implemented in 2 phases: divide the dining room and residents into smaller groups, improve traffic flow, open a cafeteria line, focus services to meet resident needs, make tray delivery and clean up more efficient, and extend meal hours. PMID- 8715753 TI - [Cardiovascular diseases. Risk factors]. PMID- 8715754 TI - [Cardiovascular diseases. Diet and prevention]. PMID- 8715755 TI - [Dietetics. Messages to be transmitted to patients]. PMID- 8715756 TI - [Cardiac insufficiency. Clinical aspects and treatment]. PMID- 8715757 TI - [Decompensated heart failure in the aged. Care by the nursing team]. PMID- 8715758 TI - [The post-phlebitis disorder. The nurse's role in the prevention of trophic problems]. PMID- 8715759 TI - [Patients with vascular diseases. Healing of skin lesions. Preparing a form on the clinical course]. PMID- 8715760 TI - [Transthoracic Doppler echography]. PMID- 8715762 TI - [Electrocardiography of effort]. PMID- 8715761 TI - [Transesophageal Doppler echocardiography]. PMID- 8715763 TI - [Veinous thrombosis. Patient education]. PMID- 8715764 TI - [Status asthmaticus at home]. PMID- 8715765 TI - [Writing in order to tell]. PMID- 8715766 TI - [Quinine. Treatment of malaria]. PMID- 8715767 TI - Valuing nursing work. PMID- 8715768 TI - Reforms round-up. Reducing infection rates. PMID- 8715769 TI - Dealing with stress at work. PMID- 8715770 TI - Reducing redundancy pain. PMID- 8715771 TI - Who are the mothers at risk? PMID- 8715772 TI - Aides need more recognition. PMID- 8715773 TI - Nursing education directions. PMID- 8715774 TI - Leaving the female body intact. AB - More than 80 million women in Africa and around the world have been damaged by the practice of female genital mutilation. As Somali refugees are accepted into New Zealand and others immigrate from African countries where the practice is still prevalent, health professionals need to be aware of what legal, ethical and cultural considerations are involved. PMID- 8715775 TI - Cutting costs and improving care. PMID- 8715776 TI - Working together on ethics. PMID- 8715777 TI - Abusing patient trust. PMID- 8715778 TI - Paving the way to discrimination. PMID- 8715779 TI - A political dilemma concerning a faculty of caring sciences in the year 2000; fact or fiction. PMID- 8715780 TI - A Swedish version of the appraisal of Self-Care Agency (ASA) scale. AB - The "Appraisal of Self-care Agency" scale (the ASA scale) is designed to measure self-care agency in accordance with Orem's self-care deficit theory of nursing. The aim of this study was to obtain a reliable Swedish version of the ASA scale by measuring the internal consistency. ASA-A for self-appraisal and ASA-B for appraisal of another were tested with a convenience sample of 52 elderly in patients and 53 caregivers in six geriatric care units. Cronbach's alpha reliability coefficient was 0.59 for the ASA-A scores, 0.77 for ASA-B scores obtained by caregivers comprising different professions, and 0.87 for scores obtained by registered nurses. Further testing of both validity and reliability is needed if this version of the scale is to be used in nursing research and practice. PMID- 8715781 TI - Influence of dinner music on food intake and symptoms common in dementia. AB - The influence of dinner music on food intake and symptoms common in dementia such as depressed mood, irritability and restlessness was studied. The study was carried out in a nursing-home ward in Sweden. Soothing music was played as dinner music for weeks, Swedish tunes form the 1920s and 1930s for two weeks and pop music for two weeks. Prior to these periods, there was one week without music, and at the end of the intervention there was a two-week control period. The effects of the intervention were assessed by psychological ratings and by weighing the food helpings. It was found that during all three music periods the patients ate more in total. The difference was particularly significant for the dessert. The staff were thought to be influenced by the music, as they served the patients more food, both main course and dessert, whenever music was played. The patients were less irritable, anxious and depressed during the music periods. The results of the study suggest that dinner music, particularly soothing music, can reduce irritability, fear-panic and depressed mood and can stimulate demented patients in a nursing-home ward into eating more. PMID- 8715782 TI - Burnout, empathy and sense of coherence among Swedish district nurses before and after systematic clinical supervision. AB - The relationships between, on the one hand, burnout, empathy and sense of cohoerence (SOC) and, on the other, personality traits were investigated, together with the effects of systematic clinical supervision on these phenomena among Swedish district nurses. The results in the supervisory group (n = 21) were compared with those of a comparison group (n = 12) in a quasi-experimental design. Personality traits were assessed by means of the Karolinska Scales of Personality. The results indicated some correlations between personality traits and burnout, empathy, and SOC, as well as correlations between the latter three phenomena. There were no significant effects of clinical supervision on burnout, empathy, or SOC. More research is needed regarding the effects of clinical nursing supervision. PMID- 8715783 TI - Nursing documentation in patient records. AB - The correct documentation of nursing care is a very important prerequisite for safe care. An extensive survey (n = 380 records), was conducted, using the NoGa protocol for a review of the nurses' documentation. The documentation revealed considerable deficiencies in most of the wards, and the nursing history, status and planned interventions were inadequate in two-thirds of the records. Furthermore, the nursing diagnosis, goals and discharge notes were especially poorly documented. The NoGa protocol was easy to use as an audit tool, useful for screening the nurses' documentation and useful for evaluation of the outcomes of educational programmes in nursing documentation. PMID- 8715784 TI - Well-being and its relation to coping ability in patients with colo-rectal and gastric cancer before and after surgery. AB - The aims of this study were, first, to describe and compare the perceived well being and general health, symptoms and coping ability of a group of patients with colo-rectal and gastric cancer before and after surgery; secondly, to describe the patients' perceptions of the hospital stay and their difficulties after discharge from the hospital; and thirdly, to investigate a possible relationship between sense of coherence and well-being. Seventy-nine (36 men, 43 women) consecutively selected patients diagnosed with colo-rectal or gastric cancer participated in the study. The Health Index (HI), the symptom checklist, the Sense of Coherence Scale (SOC scale), a study-specific questionnaire and a single item were used. Six weeks after surgery many of the cancer patients perceived that their well-being was poorer on the HI subscales energy, sleep and mobility than before. Bowel function had improved, and pain, which was a common symptom before surgery, was perceived as having lessened after surgery. Furthermore, the sense of coherence was shown to be related to the cancer patients' well-being as measured by the HI. Patients living with relatives rated their well-being as better than that of patients living alone. The problem areas identified after discharge concerned mobility, bowel function, fatigue, pain, nutrition, worry, difficulties in sleeping and problems with the wound. The instruments used in the study are seen as screening instruments to further structure the nursing-care plan, so that the patients' perceptions of the disease situation can also be taken into consideration. PMID- 8715785 TI - A nurse in preventive work--dilemmas of health information talks. AB - The possibility of making scientifically objective statements about health risks is part of a new complex of ideas pervading our Western social institutions. The concept of prevention includes an assessment of risk and dangers which lay persons have difficulties in comprehending. Advances in medical technology have made it possible to detect a growing number of anomalies in the human body and thereby to define more and more people as being at risk of various diseases. Even if it seems desirable for society to be able to detect the risks menacing its members, it calls for delicate management of the test procedures and of the preventive intervention. This article presents some perspectives on preventive work in connection with a directed health survey. The primary aim of the survey in question was to diagnose particular risk factors, for instance a high level of blood cholesterol, and inform the patient about how to prevent or reduce the risk of cardiovascular disease. The preventive measures are commonly undertaken by a nurse; her encounters with the patients--to obtain test samples, communicate the results and provide information about health risks--raise complex issues. These and other dilemmas in preventive work are discussed and analyzed. PMID- 8715786 TI - Ritualized practices among caregivers at meals in geriatric care. AB - The aim of this study was to examine the procedures of the caregivers in their daily work with meals in geriatric care, emphasizing ritualized practices, praxis and norms. An ethnographic approach was used. Registered nurses, enrolled nurses and nurse's assistants observed in the study responded to a questionnaire and, finally, were interviewed. The findings indicated that the meals mainly took the form of ritualized practices, and praxis depended on the ability and needs of the patients as well as on the working conditions. The guiding norms identified were 'to provide a home-like situation with fellowship for training purposes', 'to ensure each patient gets what he/she needs/wants by means of a fair method of serving', and 'to keep things in order and to be responsible'. These norms were transferred from the traditional family in the society, giving the caregivers the role of the parent when training patients. The enrolled nurses, responsible for the meal performance, strove to keep the eating milieu in order, and the meal as a whole was performed as a ritual. Its static form made it difficult to individualize the meal in accordance with the different needs of the patients. PMID- 8715787 TI - Nursing is not a profession that has, or does, always value leadership. PMID- 8715788 TI - Minority networks. PMID- 8715790 TI - Survival of the fittest. PMID- 8715789 TI - Leadership and you. PMID- 8715792 TI - Agenda for training. PMID- 8715791 TI - Leadership and nursing: Part One: History and politics. PMID- 8715793 TI - A manager's career path. PMID- 8715794 TI - Home front leadership. PMID- 8715795 TI - Gender in leadership. PMID- 8715796 TI - Time to act--community mental health. PMID- 8715797 TI - Leaders or managers? PMID- 8715798 TI - Service level agreements. PMID- 8715799 TI - Settling the question: Sandimmune or Neoral? PMID- 8715800 TI - Deciding on a transplant requires a close look at the benefits, drawbacks. PMID- 8715801 TI - Transplant: the next step? After 15 years on home hemodialysis, will a kidney transplant offer a better life? PMID- 8715802 TI - How an exercise program can benefit patients and the dialysis facility. PMID- 8715803 TI - Establishing training standard for dialysis technicians an important goal for NANT. PMID- 8715804 TI - Proactive approach needed to face challenges in ESRD nursing. PMID- 8715805 TI - A blueprint for re-engineering: implementing CQI in a large dialysis chain. PMID- 8715806 TI - Clinical data software in the dialysis unit: what are the essential ingredients? PMID- 8715807 TI - OIG on the warpath. Part II. When the OIG knocks on your door. PMID- 8715809 TI - Clients' health needs: nurses' concern. PMID- 8715808 TI - Nursing and the changing approach to health care. PMID- 8715810 TI - Ethical and legal issues in nursing. PMID- 8715811 TI - The midwifery nursing course: a study of problems faced by students. PMID- 8715812 TI - A day-care-centre. PMID- 8715813 TI - [Developing new concepts and ideas]. PMID- 8715814 TI - [Did you ever hear anything about ICD-10?]. PMID- 8715815 TI - [Health center hospitals: coordination, networking an interconnection]. PMID- 8715816 TI - [Case management]. PMID- 8715817 TI - [Possibilities and chances for geriatric rehabilitation]. PMID- 8715818 TI - [Let us act on behalf of our own rights]. PMID- 8715819 TI - [The importance of language in the nursing process]. PMID- 8715821 TI - [Enteral feeding, I. Advantages and problems for the patients]. PMID- 8715820 TI - [Prevention of thrombosis, Part I, II and III by Angelika Zegelin and Andreas Gerlach, "Pflege Actuell" II and 12/95 and 1/96]. PMID- 8715822 TI - [Health reform at a cross roads]. PMID- 8715823 TI - Professional development. Electrocardiography: revision notes (continuing education credit). PMID- 8715824 TI - Leading lights. PMID- 8715825 TI - No sanctuary from HIV funding cuts. PMID- 8715826 TI - Highly charged. PMID- 8715827 TI - The reluctant patient. PMID- 8715828 TI - Radical support. PMID- 8715829 TI - Ward sisters' views of the effect of NHS changes. AB - The role of the ward sister or charge nurse has long been acknowledged as being central to the provision of high-quality patient care. Health service reforms since the 1980s have led to greater managerial responsibility for ward sisters and charge nurses, who are often now known as ward managers. This paper reports on a study carried out in one hospital which described how 15 charge nurses and ward sisters saw their roles in the light of recent changes. While the informants spoke of areas of their jobs with which they were satisfied, such as control over financial resources, there were also areas of concern. These included the pace of change and the sense of isolation that they experienced. The ward sister's responses are described and recommendations for practice outlined. PMID- 8715830 TI - Leucocytes and the value of the differential count test. AB - The most frequently requested laboratory test is the full blood count. This is not one single test but a group of tests, including a count of three functionally distinct cellular elements that circulate in peripheral blood: platelets, red cells (erythrocytes) and white cells (leukocytes). This paper focuses on the function of leucocytes and the clinical significance of abnormal changes in their numbers. PMID- 8715831 TI - Prevention of complications related to contact lens wear. AB - This paper outlines how contact lenses help correct vision and identifies some ocular problems that are associated with their use. It also examines how nurses can help educate the public on issues concerning contact lenses in order to help combat ocular disease. PMID- 8715832 TI - Women's sexual health care: bringing down the barriers. AB - This paper follows the course of a health promotion project aimed at encouraging sexually active women to carry and use barrier contraceptives to maintain positive sexual health. It was carried out by four nursing students on a night diploma course at the University of Portsmouth. Having established a need and target group, an academic course project was transformed into a local community health project, thus crossing the space between education and the service sector. PMID- 8715833 TI - Is nursing anti-social? PMID- 8715834 TI - The big sleep. PMID- 8715835 TI - Wound care. Care audit cuts time and costs. PMID- 8715836 TI - Wound care. Management of leg ulcers. PMID- 8715837 TI - Wound care. Principles of Doppler. PMID- 8715838 TI - Wound care. The stigma of smell. PMID- 8715839 TI - Wound care. Trauma injury nursing in A&E. PMID- 8715840 TI - What's happened to the "care" in health care? PMID- 8715841 TI - Staff attitudes toward parent participation in the care of children who are hospitalized. AB - A nonrandom sample of 62 staff who care for children on four pediatric wards at an urban women's and children's hospital in South Australia participated in a study to identify staff attitudes toward parents participating in the care of their child in the hospital and the personal and professional characteristics that may influence these attitudes. Participants completed two questionnaires, which included the Parent Participation Attitude Scale (PPAS) and a Personal and Professional Characteristics Data questionnaire. Analysis of variance indicated that the majority of staff had a neutral attitude (PPAS score M = 72; 61-84 = neutral range) toward parents participating in their hospitalized child's care. Staff members who were registered nurses, parents, married, or associated with the burns, craniofacial, and general surgery ward or the infectious diseases ward had more positive attitudes toward parent participation than did other staff members who did not possess these characteristics. The main difference in attitude related to professional characteristics: registered nurses scored a higher mean score reflecting a more accepting attitude than any other professional group participating in the study. The results provided direction to support attitudinal and practice change focused on developing a collaborative model of care in which nurse, children, and parents work together in partnership. PMID- 8715842 TI - Easing the wait: development of a pager program for families. AB - Waiting is inevitable for patients and families seeking health care services. Anxiety, frustration, and stress are exacerbated. A pager program, where pagers are loaned free of charge to families, uses a family-centered approach to counteract the aggravation of waiting. Patients and families can leave the hospital room or clinic setting and then be paged to return as needed. In any clinical setting, having choices and control will enhance coping and satisfaction for both patients and families. PMID- 8715843 TI - A clinical case study: parent-present induction of anesthesia in children. AB - Until recently, the doors to the operating room (OR) remained closed to parents. It is well known that children undergoing surgery experience distress when separated from their parents. Like other hospitals throughout the country, Children's Hospital in Boston has responded to this distress by implementing a parent-present induction of anesthesia (PPI) program. This program has changed significantly the surgical experience for many children and families. A composite case study based on several children and parents who have participated in PPI illustrates collaboration between nursing and anesthesia staff, preparation of the parent and child, the parent's OR experience, and the generally successful outcomes of this PPI program. PMID- 8715844 TI - Ibuprofen suspension: pediatric antipyretic. AB - Pediatric ibuprofen suspension has been available by prescription for relief of fever in children age 6 months and older since 1989. A review of several studies shows this drug to be a safe and effective antipyretic in children as young as 3 months of age. Doses of 10 mg/kg have been demonstrated to provide more rapid antipyresis of longer duration than the same dose of acetaminophen, especially in the presence of high fever (> or = 101.9 degrees F). These factors prompted manufacturers to seek approval for sale of this drug without prescription. PMID- 8715845 TI - The falling grace of axillary temperatures. AB - This study compared axillary temperatures with either rectal or oral temperatures in 173 infants and young children. Axillary temperatures indicated a sensitivity of only 27.8% in their ability to detect fever in the febrile group. The age-old gold standard of estimating rectal temperature by adding 1 degree F to the oral and 2 degrees F to the axillary temperature was verified with afebrile group mean temperatures, but did not hold up with individual children's temperatures nor with febrile mean temperatures. PMID- 8715846 TI - Perceptions of Jordanian children about nurses. AB - Although children's drawings have been widely studied, a few have identified children's perceptions about nurses. Through interviews and drawings, Jordanian children were studied to examine their perceptions of nurses. These children had positive images of nurses and viewed the profession as being an attractive career choice. PMID- 8715847 TI - When cost-saving strategies are unacceptable. AB - With health care redesign, nurses in every setting are challenged to provide more cost-effective care. While quality models for health care have been developed, nurses are at times asked to implement unsafe practices in the name of cost reduction or profit building. Both legal and ethical dilemmas arise when an emphasis on economic considerations supersedes patient well-being. Nurses must recognize their primary duty as patient advocates, and work to modify unsafe plans. The ANA's Code for Nurses and Guidelines for Reporting Incompetent, Unethical or Illegal Practices, and the Nursing Practice Act are primary resources in this effort. PMID- 8715848 TI - Conducting the pediatric health history: a guide. AB - Although obtaining thorough and accurate pediatric health histories can be a challenging task, they are critically important to the health and well-being of children. A thorough history is a crucial component to both acute and episodic visits. Components of the health history are reviewed. PMID- 8715849 TI - Parent partners: a parent-to-parent support program in the NICU. Part II: Program implementation. AB - A well-organized parent-to-parent support program can provide both support and resources to help parents deal with the stress of having a baby in the neonatal intensive care unit (NICU). In order to implement a support program, hospital personnel must commit to the program, volunteer parents must be recruited and trained, and ongoing parent-professional collaboration must support program growth and maintenance. The Parent Partners Program offers a parent training curriculum and suggested procedures to establish a program. PMID- 8715850 TI - Comparisons of pain ratings from postoperative children, their mothers, and their nurses. AB - PURPOSE: To identify whether nurses and mothers of pediatric patients accurately assess the child's pain intensity as determined by the child. METHOD: A descriptive correlational design examined the perception of pain from a nonrandom convenience sample of 20 postoperative school-aged children, their mothers, and their nurses. A 100 mm visual analogue scale was completed by all participants at 3 separate observations. Pearson's Product Moment Correlation was used to analyze relationships between the dyads. FINDINGS: The mother/child dyad showed significant relationships at observations 1 and 2; these correlations were higher than other correlations at each observation. There were significant relationships for the child/nurse dyad at observations 1 and 2, and for the mother/nurse dyad at observation 1. CONCLUSIONS: Mothers may be a valuable source of information in assessing their child's pain. The use of pain assessment tools may need to be examined and incorporated into nurses' practice, and assessments of children's pain may need to integrate more data pertaining to developmental level. PMID- 8715851 TI - Pediatric management problems. What is your assessment? Scarlet fever. PMID- 8715852 TI - One year later: a look at the contract with America. PMID- 8715853 TI - Children's services still inadequate--37 years on. PMID- 8715854 TI - Children first: the local audit. PMID- 8715855 TI - Do siblings need a special day? PMID- 8715856 TI - Changes in Russian nurse education. PMID- 8715857 TI - Imperforate anus: nutritional care. PMID- 8715859 TI - Vaseline gauze dressings after circumcision. PMID- 8715858 TI - Failure to thrive in children with chronic illness. PMID- 8715860 TI - Children with cancer: managing stress in staff. PMID- 8715861 TI - Pain assessment and the use of morphine. PMID- 8715862 TI - Midwifery practice: revolution or evolution? PMID- 8715863 TI - Ageism: the challenge for nursing. PMID- 8715864 TI - Depressed elderly: who cares? PMID- 8715866 TI - Quality care in crisis--the American experience. PMID- 8715865 TI - Asthma & elderly people. PMID- 8715867 TI - Managed care and case management. PMID- 8715868 TI - Weaning from mechanical ventilatory support. AB - Weaning patients from mechanical ventilatory support continues to be a major challenge in critical care units. This article discusses recent research in the area of weaning and identifies specific parameters related to successful weaning in clinical practice. PMID- 8715869 TI - Caring for the ICU patient receiving propofol. AB - This article, the first in the new DCCN department for acute-care nurse practitioners, clinical nurse specialists, and advanced practice nurses, focuses on the pharmacodynamics of propofol. The advanced practice nurse working in the ICU is likely to institute propofol protocols, teach staff about the drug, and collaborate with the bedside nurse on the patient's outcome to this treatment. PMID- 8715870 TI - Characteristics related to DNR orders for pediatric ICU patients. AB - Although technology exists to sustain vital functions of almost any patient, not every pulse-less patient is a candidate for resuscitation. Criteria needs to be developed to guide health care professionals in identifying DNR situations and teaching families about DNR options. The first step is to identify characteristics that are related to current DNR selection, so prescriptive criteria can be developed to aid in DNR decisions in the future. This article presents a descriptive study of characteristics associated with DNR statistics for pediatric ICU patients. PMID- 8715871 TI - Achieving collaboration in ethical decision making: strategies for nurses in clinical practice. AB - Ethical decision making is a process that combines justice and caring in moral reflection to select sound choices. Nurses and physicians often have different perspectives on how to resolve ethical dilemmas. Satisfactory resolution depends on overcoming conflict and achieving collaboration between members of the health care team. Conflict can occur on a number of different levels. For example, it can be between nurses, nurses and physicians or the entire health care team and the patient. This article helps prepare critical care nurses to handle ethical dilemmas in crisis situations by providing them with specific strategies that help promote collaboration in resolving ethical dilemmas. PMID- 8715873 TI - A new role for critical care nursing: the acute care nurse practitioner. PMID- 8715872 TI - Adapting a thesis to publication style: meeting editors' expectations. AB - Every year hundreds of thesis reports are completed by graduate students. Many of these end up on nursing journal editors' desks, only to be rejected. What characteristics of a thesis lead to rejection? How can authors develop quality research manuscripts? How can faculty teach graduate students the difference between a thesis and manuscript. This article answers these questions by providing advice from 15 critical care and research journal editors. PMID- 8715874 TI - Continuing education: continuing competence. PMID- 8715875 TI - Factors associated with critical thinking among nurses. AB - The ability to think critically is increasingly hypothesized to play an important role in a nurse's ability to function competently in complex health care settings. This cross-sectional exploratory survey used secondary data (Gendrop, 1989) to assess the critical thinking ability of a convenience sample of 160 nurses from two urban hospitals. Four variables hypothesized to be associated with critical thinking ability were explored: age, level of education, years of nursing experience, and area of expertise in nursing. Critical thinking was assessed using the Watson Glaser Critical Thinking Appraisal Tool (WGCTA). Study findings suggest that age and years of experience were negatively correlated with critical thinking ability. However, the level of education was positively related to critical thinking. Future needs for research were identified and implications for nursing education and practice were provided. PMID- 8715876 TI - Pathway to excellence: a peer based program in continuing education. AB - Maintaining the professional knowledge and competency needed to practice in today's acute care settings is becoming increasingly difficult. This article introduces a continuing education program developed to assist staff nurses in meeting their learning needs. "Pathway to Excellence: Continuing Professional Education" is a peer education program or nursing education for nurses, by nurses, that can be implemented in all areas of clinical practice. PMID- 8715877 TI - Creative teaching strategies. AB - Today's nursing students feel pressured to complete research-based professional papers. Reasons that these assignments cause feelings of anxiety may be due to learners' poor writing/thinking skills, low literacy levels, lack of attention when traditional strategies are used, or expectations of entertaining teaching methods. Creative teaching strategies, using a story-telling and personification approach, may assist nurse educators to improve students' attention, retention and comprehension of primary concepts in their nursing courses. PMID- 8715878 TI - Assessment of a continuing education evaluation framework. AB - This study analyzes a systems model for evaluation research of continuing education (CE) and reports on the result of a quasi-experimental study of a stoma care CE program. Holzemer's conceptual framework for evaluating CE consisting of the learner, setting, and program along with a systems model was used as an organizing framework. A quasi-experimental design (n = 24 experimental group and n = 27 control group) was used to evaluate the impact of the CE program. Pre- and post-tests of participants' knowledge and skills of stoma care were evaluated. Participants' satisfaction with the course was surveyed by questionnaire during the course, on completion and four months after the course. No significant differences were found between the two groups. The revised framework included the concept of needs assessment and the impact of the CE program on patient care. PMID- 8715879 TI - Enculturation of foreign nurse graduates: an integrated model. AB - Graduates of foreign nursing schools are a unique and valuable resource of the U.S. health care delivery system. Due to inadequate communication skills and a lack of cultural enculturation of many foreign nurse graduates (FNGs), some may not reach their full potential as a professional nurse. Agencies who employ FNGs can greatly improve their integration by providing continuing education to enhance communication and cultural enculturation. This model of a communications skills course addresses all aspects of communication: therapeutic, verbal and non verbal, while integrating a cultural component. Thus, the implementation of this model can enhance foreign nurse graduates' ability to practice in the U.S. health care system. PMID- 8715880 TI - Nurse administrators' perceptions of the impact of continuing nursing education in underserved areas. AB - Qualitative inquiry can provide new dimensions to existing knowledge about how nurses use information from continuing education participation in practice. This study offers descriptions from the perspective of nurse administrators in an underserved area and a time of shrinking resources of nurses' application and use of knowledge gained from continuing education participation. Summarized within a model of change, learning was expressed in personal and professional dimensions including stimulus for innovation, educational and career mobility, networking, sharing knowledge, problem-solving, and improving patient care. Change indicators were the infrastructure linking the implementation in professional practice. PMID- 8715881 TI - Nonpharmacologic interventions to manage children's pain: immediate and short term effects of a continuing education program. AB - A two-group pretest, posttest design was used to test the effects of a two-hour educational program about using five nonpharmacologic pain management techniques with children on nurses' knowledge of, comfort with, attitude toward, and use of the discussed techniques. Five pediatric patient care units were randomly assigned to a treatment or control group. Pretest responses from 54 nurses indicated that attitude was the only significant determinant of their intention to guide children's imagery. A subset of 35 nurses actually attended the program and completed posttest responses. On posttest, the treatment group reported significantly greater knowledge of and comfort with using the techniques. Comparison of pretest with posttest responses across both groups indicated a significant increase in knowledge of and comfort with using nonpharmacologic techniques. Two months after the program, 24 nurses from both groups reported increased use of the techniques in practice compared with prior to the program. PMID- 8715883 TI - Opposition health spokesperson: Ms Wendy Edmond. PMID- 8715882 TI - Queensland's new Health Minister: Mr Mike Horan. PMID- 8715884 TI - This is an emergency. PMID- 8715885 TI - Rural community nursing. PMID- 8715886 TI - Infectious disease control: midwifery services and obstetric procedures. PMID- 8715887 TI - Coincidental guidance. PMID- 8715888 TI - Musings on the AHMA/AHNA conference: the doctor & nurse perspective. PMID- 8715889 TI - Surfing the holistic wave: resources on the Internet for health & healing. PMID- 8715890 TI - Imagery & magic: new AHNA program brings vision to life. PMID- 8715891 TI - Passage meditation and the use of a mantram. PMID- 8715892 TI - [Analysis of relative factors on paralytic patients with pressure sore complications]. AB - In this study, risk factors for bedsore were studied on 100 paralytic patients with neurologic diseases. The patient's assessment scores based on the risk factors for bedsore were evaluated and analyzed. According to the result, the critical point of assessment score for bedsore was established. The patient will be put into the high risk group of bedsore if his/her assessment score is higher than the critical point and special nursing intervention will be given. PMID- 8715893 TI - [Care of alleviating therapy of nephrotoxicity due to high-dose cisplatin chemotherapy]. PMID- 8715894 TI - [Nursing care of direct microsurgery of intracranial aneurysm]. PMID- 8715895 TI - [Care of pericardial biopsy with indwelling catheter drainage]. PMID- 8715896 TI - [Care of induction of labor by intravenous drip of oxytocin]. PMID- 8715897 TI - [Study of negative pressure drainage fluid and the fluid cellular ingredients after laryngeal carcinoma radical neck dissection]. PMID- 8715898 TI - [Nutritional care of severe burn patients during the hypermetabolic stage]. PMID- 8715899 TI - [Care of plastic surgery of neck and face during awakening stage of general anesthesia]. PMID- 8715900 TI - [Care of children after percutaneous balloon pulmonary valvuloplasty]. PMID- 8715901 TI - [Analysis of the causes of 230 cases of neonatal asphyxia]. PMID- 8715902 TI - [Clinical study of digital blood pressure monitor]. PMID- 8715903 TI - [Quantifying the information of nursing management for prediction of nursing work]. PMID- 8715904 TI - [The application of change theory on nursing administration]. PMID- 8715905 TI - [Teaching nursing theory and acculturate the person]. PMID- 8715906 TI - [Care of catheter radio frequency in children]. PMID- 8715907 TI - [Analysis of the articles published in the Chinese Journal of Nursing from 1989 1994]. PMID- 8715908 TI - [Battle against pressure ulcers]. PMID- 8715909 TI - [A plan of teaching and research. Ciudad Sanitaria Universitaria de Bellvitge]. PMID- 8715911 TI - [Fetus in the uterus in the 5th and 6th months]. PMID- 8715910 TI - [Health and the woman. Menopause: evaluation of a health program]. PMID- 8715912 TI - [Preventive measures. The risk of hypothermia during prolonged anesthesia and surgery. Study of methods of warming]. PMID- 8715913 TI - [Voluntary intoxications that require hospitalization. Study at the Navarra hospital]. PMID- 8715914 TI - [Transparent hydrocolloid dressings for neurosurgical wounds]. PMID- 8715915 TI - [Evaluation of a convex two-piece ostomy device]. PMID- 8715916 TI - [Incontinence in a home for the aged]. PMID- 8715917 TI - [Backache in nurses. What is the cost to the system?]. PMID- 8715918 TI - [Habeas corpus--give me my body!]. PMID- 8715919 TI - [Demand supervised care]. PMID- 8715920 TI - [No research without ethics]. PMID- 8715921 TI - [Incontinence--the practical side. Incontinence seen as a nursing problem]. PMID- 8715923 TI - [Yes to life-saving interventions in the elderly]. PMID- 8715922 TI - [Confusion about sucking behavior--a study in 3 countries. Don't confuse the baby!]. PMID- 8715924 TI - [The nursing student makes a mistake. Who is responsible?]. PMID- 8715925 TI - [At the service of patients with stomas: the stoma therapist]. PMID- 8715926 TI - [Allergy to natural latex: how to prevent it?]. PMID- 8715927 TI - [Reflections about the changes in our society. 2. Liberating the future]. PMID- 8715928 TI - [Ethical dilemmas in intensive care: when one has to chose between loyalty and charity]. PMID- 8715929 TI - A fight for childbearing women to be treated with dignity and respect. PMID- 8715930 TI - Obstetric care: competition or co-operation. AB - OBJECTIVE: The aim of this study was to determine the feasibility of co-operation within maternity and obstetric care between midwives, general practitioners (GPs) and obstetricians. DESIGN: descriptive correlational study. SETTING: The Netherlands. Policy is towards more co-operation between primary and secondary health care. However, in Dutch health care midwives, GPs and obstetricians may also have conflicts of interests. PARTICIPANTS: members of obstetric co-operation groups (91 midwives, 53 GPs, 31 obstetricians) completed a questionnaire. MEASUREMENTS AND FINDINGS: in the questionnaire information was collected on what members of obstetric co-operative groups expect from co-operation. Findings indicated that consensus existed about experienced advantages and disadvantages of co-operation, tasks that should be achieved, and how obstetric co-operative groups could be set up. Nevertheless, there was evidence of competition and there were also conflicting ideas about co-operation. KEY CONCLUSIONS AND IMPLICATIONS FOR PRACTICE: it is concluded that the findings have implications for the organisation of an obstetric co-operative group. If professionals want to start such a group it is preferable to start with topics that benefit all participants (win-win situation) and motivate them to participate actively. A second step may be an attempt to reach agreement about how to communicate with each other in the case of referrals and consultations. During this phase mutual trust and respect may grow, so that finally more difficult problems can be discussed (mixed-motives situation). PMID- 8715931 TI - Women's experience of the encounter with the midwife during childbirth. AB - OBJECTIVE: to describe women's experience of the encounter with the midwife during childbirth. DESIGN: a qualitative study using a phenomenological approach. Data were collected via tape-recorded interviews. SETTING: the Alternative Birth Care Centre, Sahlgrenska University Hospital, Gothenburg, Sweden in 1994. PARTICIPANTS: 18 women, six primiparous and 12 multiparous who were two to four days post delivery. KEY FINDINGS: the essential structure of the studied phenomenon was described as 'presence' and included three themes: to be seen as an individual, to have a trusting relationship and to be supported and guided on one's own terms. IMPLICATIONS FOR PRACTICE: the need to be seen as an individual can be realised by affirmation and familiarity with the midwife and surroundings. A trusting relationship can be obtained by good communication and proficient behaviour. By providing a sense of control the women can be supported and guided on their own terms. Above all they must feel that the midwife is present. PMID- 8715932 TI - Pre-registration midwifery programmes: a case study evaluation of the non midwifery placements. AB - OBJECTIVE: to evaluate the effectiveness of non-midwifery placements in enabling pre-registration (direct entry) student midwives to learn about caring for childbearing women with medical, surgical or mental health problems and needs. DESIGN: case study. SETTING: a large midwifery education department and three acute general hospitals in England. PARTICIPANTS: 15 student midwives in the first intake of one college's three-year diploma programme in midwifery plus the practitioners involved in their education. MAIN OUTCOME MEASURES: the learning needs of student midwives, who have no previous health-care experience, to enable them to care effectively for women with medical/surgical/mental health problems; the teaching processes and learning opportunities related to these aspects of care; how the views of teachers and practice placement staff compare with those of the students; the factors that influence the teaching and learning processes and experiences for pre-registration student midwives. FINDINGS: the broad range of experiences in medical/surgical/metal health placements enabled students to increase in maturity, and confidence and develop their communication skills. The variety of placements enhanced student understanding of the multi-disciplinary team's contribution to health care, students learned new practical skills which were transferable to maternity care contexts and all students had opportunities to care for adults with most of the medical/surgical/mental health problems seen in childbearing women. CONCLUSION: this action research project provided data for curriculum development and helped to avoid premature reaction to individual staff and student response. The value of the medical/surgical/mental health placements and the importance of staff and student preparation for effective learning was established. Whilst it might be essential to identify what and where student midwives should learn, it would appear that developing each student must be an equal, if not greater, priority for curriculum designers. PMID- 8715933 TI - Midwives' and senior student midwives' knowledge of haemaglobinopathies in England. AB - OBJECTIVE: to examine midwives' and senior student midwives' knowledge concerning sickle-cell anaemia and beta-thalassaemia. DESIGN: survey using the 'Dyson' questionnaires. SETTING: study days on 26 sites across England over three months. PARTICIPANTS: 850 questionnaires were completed by a convenience sample of midwives and senior students: 401 on sickle-cell anaemia and 449 on beta thalassaemia. In each case just under two-thirds were qualified midwives. MEASUREMENTS AND FINDINGS: most respondents underestimated the number of ethnic groups affected by these disorders. A majority gave the wrong answers to basic questions on the inheritance of these disorder, fewer than one-third answered two simple genetics questions correctly. KEY CONCLUSIONS: in order to provide an equitable service midwives need more education in this area, particularly after qualification. IMPLICATIONS FOR EDUCATION PRACTICE: education on haemoglobinopathies was linked with improved knowledge levels and appears most effective when undertaken by counsellors in haemoglobinopathies. Such education should therefore be available, especially to qualified midwives who had not previously received it. PMID- 8715934 TI - Marketing midwifery education: findings from a survey. AB - OBJECTIVE: to collect information from a sample of identified customers (midwives and midwifery managers employed by health authorities and trusts) in order to develop a marketing strategy for a Department of Nursing, Midwifery and Health Care. DESIGN: two descriptive and analytical surveys using questionnaires. SETTING: mid- and west Wales, UK. SUBJECTS: randomly selected sample of 75 midwives and census sample of eight midwifery managers working within the National Health Service. MEASUREMENTS AND FINDINGS: quantitative and qualitative data collection methods. KEY CONCLUSIONS: organisations have to define, and listen carefully, to their customers and offer courses which are appropriate, related to clinical midwifery practice and have titles that accurately reflect the content. Midwives are enthusiastically committed to continuing education despite the current difficulties. Word of mouth and personal recommendation remains the most effective form of advertising. University departments of nursing and midwifery can easily become removed from clinical practice. Education assists practitioners in changing and adapting to a new order; it is also crucial in developing skills in critical thinking and analysis. New skills are needed if practice is to improve. IMPLICATIONS FOR PRACTICE: academic departments can easily become remote and out of touch with the needs of clinicians and midwifery managers. Departments must take steps to define the customer, listen carefully to what they want and make every effort to provide continuing education for midwives that is relevant, responsive, accessible and attainable. The benefits of education also must be marketed. PMID- 8715935 TI - Conclusions of a round-table that took place during a seminar on the prevention of TB and HIV transmission in health care settings, Douala, Cameroon, January 1995. PMID- 8715936 TI - Competencies: a tool for continuous learning and change. PMID- 8715937 TI - Tips for creating a customer listening system. PMID- 8715938 TI - Information technology. PMID- 8715939 TI - [Study on preventing alopecia caused by chemotherapy with cold pillow compresses]. AB - Thirty cancer patients were studied by using cold pillow compress to prevent alopecia resulting from chemotherapy. The result indicated that persistent use of cold pillow compress, which made the patients comfortable, could reduce the hair follicles inhibition or damage which caused by chemotherapeutic agents. So alopecia can be prevented or decreased. PMID- 8715940 TI - [Care of posture during trans-cutis trans-pedicle vertebral external fixation of the thoracic-lumbar spine]. PMID- 8715941 TI - [Nursing of implantation of prosthesis of whole scapula and shoulder joint]. PMID- 8715942 TI - [Care of continuous passive motion after semi-joint reconstruction]. PMID- 8715943 TI - [Studies on methods of venipuncture for patients with traumatic and hemorrhagic shock]. PMID- 8715944 TI - [Care of bleeding of acute radiation disease]. PMID- 8715945 TI - [Care of stubborn simple nephrosis using pharmacotherapy in children]. PMID- 8715946 TI - [Care of the repairing of periodontal bone defects with ceramic bovine bone compounded with bBMP]. PMID- 8715947 TI - [Care of internal plombage with inert gases to treat old retinal detachment]. PMID- 8715948 TI - [Survey of heterophoria of 1500 normal male youth]. PMID- 8715949 TI - [Nursing care of pregnant and parturient patients associated with intracerebral hemorrhage]. PMID- 8715950 TI - [The design and implementation of nursing care plans]. PMID- 8715951 TI - [Developing and applying of the microcomputer managing system for nursing]. PMID- 8715952 TI - [The formation of head nurses' influence and in practice]. PMID- 8715953 TI - [Teaching strategies in conducting nursing psychology courses]. PMID- 8715954 TI - [Progress of epinephrine in resuscitation of the heart]. PMID- 8715955 TI - [Introduction to the assessment methods of the at-risk pressure sore patient]. PMID- 8715956 TI - [Nursing courses in Germany. 1. The diversity of expectations causes problems]. PMID- 8715957 TI - [Nursing seminar in Ulm: "I would take part again anytime"]. PMID- 8715958 TI - [Self care for nurses. 1. Getting fit for spring]. PMID- 8715959 TI - [Nosocomial urinary tract infections: the drainage system has a key position]. PMID- 8715960 TI - [Pain monitoring: only the joint visit by physician and nurse is meaningful]. PMID- 8715961 TI - [Congress: "Alternative healing methods"; great interest in compresses and bandages]. PMID- 8715962 TI - [Caring for oncological patients: transitional care simplifies the transition from hospital to home]. PMID- 8715963 TI - [An exchange of letters with Bonn makes it clear: nurse administrators must not also run the nursing school]. PMID- 8715964 TI - [Ethical limits in intensive care: don't unnecessarily prolong the dying process]. PMID- 8715965 TI - [Quality assurance on the ward. Increasing their knowledge increases the nurses' self esteem]. PMID- 8715966 TI - [Marketing in care: is humaneness being driven out of the hospital?]. PMID- 8715967 TI - [Federal decree in nursing economics 1995. 1. Economic problems are moving toward center]. PMID- 8715968 TI - [Reimbursement of course fees: employers have poor cards]. PMID- 8715969 TI - [Improvement of student instruction: a manual assures the interlocking of theory and practice]. PMID- 8715970 TI - [Occupational disorders of nursing personnel. 4. Nurses are particularly stressed during night shift]. PMID- 8715971 TI - [Professionalizing nursing: how would you like the term "health expert"?]. PMID- 8715972 TI - [Nurses between vocation and profession]. PMID- 8715973 TI - [An instrument for better dealing with costs: minimal data for nursing care]. PMID- 8715975 TI - [New federal health insurance legislation. What is changing for private practice ?]. PMID- 8715974 TI - [Pharmaceutic assistance and interprofessional management of health costs. An experience in Fribourg]. PMID- 8715976 TI - [Economic evaluation of health interventions. Spending health money: making good choices]. PMID- 8715977 TI - [January 21, 2010: the day the world exploded]. PMID- 8715978 TI - [The powerlessness of nurses--and their power. Power as a chance]. PMID- 8715979 TI - [With money, man immortalizes himself on this earth. The dangerous magic of money]. PMID- 8715980 TI - [Conjectures on the effects of the new health insurance legislation. The health insurance legislation and the Gretchen problem]. PMID- 8715981 TI - [Profitability is influencing decisions. We must not calculate costs only but also the results]. PMID- 8715982 TI - ['Floor nursing' at the start of rehabilitation--report of experiences. Mattress camping for the brain damaged]. PMID- 8715983 TI - [Just talking]. PMID- 8715984 TI - A video worth a thousand words. PMID- 8715985 TI - The benefits of Tai Chi. PMID- 8715986 TI - Caring touch: for the frail, elderly and dying. PMID- 8715987 TI - Certificate program in holistic nursing. Healing touch schedule. PMID- 8715988 TI - [Head and brain injury--clinical picture and diagnosis]. PMID- 8715989 TI - [Nursing aspects in brain injuries]. PMID- 8715990 TI - [Interviewing parent at a neonatal intensive care unit--a project from practice]. PMID- 8715991 TI - [Nursing tasks in surfactant therapy for premature infants and newborns]. PMID- 8715992 TI - [Nosocomial infections in pediatrics]. PMID- 8715993 TI - [Viral hepatitis--still a current topic]. PMID- 8715994 TI - [Dialog with the physician--between helplessness of the participants and desirable partnership]. PMID- 8715995 TI - [The introduction of nursing standards]. PMID- 8715996 TI - [Toxic and less toxic plants. 24]. PMID- 8715997 TI - [From school to an artificial kidney--from the everyday life of a child in dialysis treatment]. PMID- 8715998 TI - [Cystic fibrosis--increasing life expectancy of patients with cystic fibrosis, supported by data]. PMID- 8715999 TI - [Dirty hands. Bulletin of the European Association for the Promotion of Hand Hygiene]. PMID- 8716000 TI - [New rules for working conditions--the important contractual conditions must always be in writing]. PMID- 8716001 TI - [The war, consequences and no end--traumatism in wartime children]. PMID- 8716003 TI - "Is there a future for nursing"? PMID- 8716002 TI - [Vaccines are biologicals that need care]. PMID- 8716004 TI - Nursing students see light at end of health care reform. PMID- 8716005 TI - Publication of nurse conduct hearings--lifting the shroud of secrecy. PMID- 8716006 TI - Understanding the effects of torture. PMID- 8716007 TI - Diversity in nursing has its merits. PMID- 8716009 TI - Continuing competence: a constant challenge. PMID- 8716008 TI - Clinical practice guidelines. Registered Nurses Association of British Columbia. PMID- 8716010 TI - Code of ethics review. PMID- 8716011 TI - Patient care delivery systems: implications for nursing practice. PMID- 8716012 TI - Alternative/complementary therapies. PMID- 8716013 TI - How nurses can help supporting victims of torture. PMID- 8716014 TI - The reuse process: a continuous quality improvement project. PMID- 8716015 TI - A look at worldwide use of peritoneal dialysis. PMID- 8716016 TI - Be aware: OIG, other agencies bearing down on renal care, health care industry. PMID- 8716017 TI - The monthly capitated payment/Part II. Establishing relative work values, looking at practice expenses. PMID- 8716018 TI - Taking capitation to the nephrologist: an update on the RPA project. PMID- 8716019 TI - Capitol views. Capitation: will it work for ESRD? PMID- 8716020 TI - Nurses, technicians face the future. At the beginning: how we created the NNTCB exam. PMID- 8716021 TI - Nurses, technicians face the future. Case management: where ESRD nursing is heading. PMID- 8716022 TI - [Paragraphs on decubitus ulcers--the legal situation with reference to decubitus ulcers]. PMID- 8716023 TI - [Autonomy, definition and legal consequences]. PMID- 8716024 TI - [Potentials of the new wound treatments]. PMID- 8716025 TI - [Skin]. PMID- 8716026 TI - [Aroma therapy--a new pathway in health care and nursing]. PMID- 8716027 TI - [Decubitus risk: how important are risk scales?]. PMID- 8716028 TI - [A useful item: a new method for the acceleration of wound healing]. PMID- 8716029 TI - [Nursing education--somewhat different]. PMID- 8716030 TI - Three nursing home patients who, due to inadequate care and poor nutrition, developed severe bed sores. PMID- 8716031 TI - Using a gentler hand: reflections on patients with pressure ulcers who experience pain. AB - Nurses who care for patients with pressure ulcers who experience pain were asked to reflect and write a story about this phenomenon. The objective of this study was to determine the meanings derived from nurse caregivers' reflections about patients' pressure ulcer pain. A Heideggerian hermeneutical approach was the methodology used for this inquiry. Stories from 42 participants, including nurse generalists and advanced practice nurses, were analyzed. Input was solicited from two qualitative research teams. Findings consist of three constitutive patterns and eight related themes: Nursing expertly (reading the pain, attending to the pain, and acknowledging and empathizing); Denying the pain (assuming it doesn't exist, not hearing the cries, and avoiding failure); Confronting the challenge of pain (coping with the frustrations and being with the patient). Lessons learned from this analysis are to stay connected with your patients; listen, attend ("be with") and comfort; and use a gentler hand. PMID- 8716032 TI - Breaking the cycle: the etiology of incontinence dermatitis and evaluating and using skin care products. AB - Dermatitis is a common problem for incontinent patients; a cycle of moisture, friction, bacteria and breakdown can be established. To break this cycle, the clinician must understand the etiology of incontinence dermatitis and choose appropriate skin care products. Healthy skin has a mean acid mantle of 5.5 pH. This natural acidity discourages bacterial colonization and provides a moisture barrier. Aging skin experiences increased dryness (allowing cracks in which bacteria colonizes) and slower recovery from the effects of alkaline substances. Incontinence results in elevated friction coefficient, exposure to moisture, bacteria and ammonia (leading to alkaline conditions), and increased enzymatic activity. When choosing products to protect the skin, clinicians should check that the product's pH level is within the desirable range (4 to 7), that ingredients are appropriate and not contraindicated for particular patients, and that the product has been tested for dermal irritation and antimicrobial efficacy. Three basic product types are cleansers, moisturizers and barriers. Skin should be cleansed gently without high alkalinity (as in some bar soaps) or rubbing (which denudes fragile skin), patted or air dried, moisturized after bathing, and protected with a barrier. Using the appropriate products can reduce the risk for breakdown and enhance the skin health of your patients. PMID- 8716033 TI - Arterial ulcers: assessment and diagnosis. AB - Arterial ulcers occur because of inadequate perfusion of skin and subcutaneous tissue at rest. Arterial occlusive disease, common among smokers, diabetics and the elderly, can lead to claudication, rest pain and gangrene, in addition to localized ulceration. Other processes, such as venous stasis, pressure, trauma, and vasculitis, can also cause ischemia. However, a thorough patient history and physical assessment can help discriminate between ischemic ulcers caused by arterial disease and other types of ulcers. The key to the diagnosis of arterial occlusive disease is the patient history. Pain while walking is the most common presenting complaint and can indicate intermittent claudication. Physical assessment should include both a general exam, looking for problems relating to lungs, heart and nervous system, and a focused exam of the affected extremities and arterial pulses. Vascular laboratory findings can also help confirm a diagnosis of arterial ischemic ulceration. The key to treatment is improvement in the vascular perfusion to the affected area. Surgical revascularization is the mainstay of treatment, with some interventional procedures becoming accepted. Medical options, in addition to correction of underlying medical problems, include good wound and supportive care, but pharmaceutical interventions have generally not proven effective, and should be considered only if interventional procedures are not possible. With an adequate blood supply reestablished, most arterial ulcers will progress to healing unless there are complicating factors. PMID- 8716034 TI - Strategic planning for the wound care clinic in a managed care environment. AB - The present trend in healthcare delivery is away from classic fee-for-service practices toward providing tightly controlled "products" in the form of Health Maintenance Organizations (HMO's). To thrive in this changing marketplace, wound clinic practitioners must understand the background of managed care and its terms, and be familiar with the concepts of capitation and incentives. Part of managed care's impact upon the specialty of wound care is that practitioners must define outcomes, healing rates and standards of care. Patient referral will depend on the cost effective delivery of care from all specialists in addition to the primary care physician. To meet these needs the authors created a hospital based wound care clinic with multiple interrelated areas of influence and have gathered statistical data. Managed care is here to stay and wound care practitioners must learn how to prepare for it and deal with it effectively. PMID- 8716035 TI - Collaboration in wound care: a nursing and physical therapy team approach. PMID- 8716036 TI - Managed care and wound care: proactive strategies for successful outcomes. PMID- 8716037 TI - Draft guideline: diagnosis and treatment of venous leg ulcers. PMID- 8716038 TI - The alluring world of cigars. PMID- 8716039 TI - Suggested enuresis treatment regressive. PMID- 8716040 TI - Surveying infant feeding practices. PMID- 8716041 TI - [Know how and self knowledge]. AB - This study aimed to optimize the education process by bringing about changes in the evaluation practices of nurse educators based upon changes in their representations of knowledge, know-how and self-knowledge. Results indicated that changes are more likely to occur among those teaching relation subjects and with previous institutional experience. PMID- 8716042 TI - Canuck place: a hospice for dying children. AB - Parents will never face a more stressful event than the news that their child will die. A child's terminal illness challenges every belief, emotion and dream that a parent has. In addition to the emotional drain, caring for a dying child is physically exhausting. Moreover, the child's illness affects the lives of all brothers, sisters, aunts, uncles and grandparents. The care of these children and their families demands a holistic program of services, including physical and emotional support, as well as tangible ways to relieve parents from the continuous 24-hour care of their child. And after the child has died, the emotional support must continue as families learn to face life without their child. PMID- 8716043 TI - Staff nurses' perspectives of hospital power structures. AB - After almost 20 years of working and teaching in nursing, I am left wondering why physicians get all or most of the credit for successful patient outcomes, yet nurses frequently do most of the work--a fact that goes quietly unrecognized. Then there are the puzzling queries I hear repeatedly from students during their clinical experiences. They question why nurses seem unclear about a common identity--what it is they are doing that is unique to their profession. They also claim that nurses in conflicts often side with other health care providers or management, instead of supporting each other. PMID- 8716044 TI - Premature infants and car seat safety. AB - Automobile accidents are one of the leading causes of mortality and morbidity in children. Indeed, the rate is increasing, even though most other causes of child death have decreased in the last five years. The misuse and non-use of child restraints are at the root of this problem. PMID- 8716045 TI - Alternative cancer therapies. AB - Conventional treatments for cancer are designed to cure the disease or slow its destructive effects, but they do little to establish a feeling of control in the patient. Because of the nature of the disease, patients often consent to treatment with the frightening knowledge that a cure cannot be promised. In addition, the side effects of surgery, chemotherapy and radiotherapy can make patients very ill, both physically and psychologically. PMID- 8716046 TI - [The existential meaning of time for the aged]. AB - The notion of time is particularly important to seniors. Because their future prospects are more limited, the past plays a very significant role. It serves as an anchor for the present and a guideline for the future. The passage of time is not uniform for everyone. It evolves over years and is based on accumulated experiences and personal factors such as age, activity and interests. For this reason, the older a person gets, the more each minute counts. Time is also indicative of change and is revered as the value of values. It is a useful tool for highlighting and valuing the present moment and the actual task to be accomplished in the here and now. It is essential that nurses understand the subjective reality of time to each individual patient if they are to give quality care. Comprehending time's existential meaning to seniors will add a further dimension to nursing interventions that are based on Watson's model of care. PMID- 8716047 TI - [Aging: better understanding, better care]. AB - Although aging is a developmental stage, bias against seniors is still pervasive. This phenomenon is not new; history has recorded a devaluing of the elderly. Perceptions of old age and aging also affect the delivery of health care and nursing services. A nurse's pessimistic viewpoint can translate into a loss of dignity, identity and decision-making power for seniors. It is essential that nurses use experiential and scientific knowledge to promote a better understanding of the aging process as part of normal development. These two types of knowledge will enable them to understand the process in its entirety and to grasp the true meaning of being old. Chronological age is not the only clue to biological aging. There is no unique model. Aging is a combination of various profiles of complex relationships between biological, psychological and social variables. A better knowledge of the normal aging process will encourage nurses to have a realistic and positive view, to facilitate elders' adjustment, and help them to maintain a better quality of life. PMID- 8716048 TI - Facsimile transmissions. AB - Some provincial regulators of nursing are setting out professional standards for the use of facsimile transmissions. While the facsimile machine is a tool that can improve client care through the timely, accurate transmission of vital information, nurses should recognize the potential hazards. Clear policies and procedures for the usage and management of facsimile transmissions are necessary to ensure that legal and professional standards are met. PMID- 8716049 TI - Community "wise" women. PMID- 8716050 TI - Putting control theory to work. PMID- 8716051 TI - The user fee debate. AB - The issue of user fees for health care generates a highly charged and emotional response from most Canadians. Access to health care services based on need rather than income is a feature of our system, of which we are extremely proud. Yet, there is increasing pressure on provinces to consider user fees. Are user fees the best method to cope with these financial pressures? Nurses can play a decisive role in ensuring that governments and the public are continuously well informed on this issue. PMID- 8716052 TI - [Nurses' patient transfer report. Concept for the determination of the nursing transfer status at a surgical intensive care unit]. PMID- 8716053 TI - [Rare urological tumors]. PMID- 8716054 TI - [After care and functional control in patients with pacemakers]. PMID- 8716055 TI - [Infections in surgery patients--etiology, diagnosis, prevention. 2]. PMID- 8716056 TI - [The depressive patient. Important points in nursing care]. PMID- 8716057 TI - [Personal hygiene in confused elderly. "More than cleanliness and freshness..."]. PMID- 8716059 TI - [Communication model. Wolf or giraffe--that is the question]. PMID- 8716060 TI - Confidentiality, human genetics and the OH nurse. PMID- 8716061 TI - Maintaining first aid skills, a question of time? PMID- 8716062 TI - Practise makes perfect: disaster management. PMID- 8716063 TI - Work and ageing. PMID- 8716064 TI - Disability Discrimination Act 1995: an occupational health nightmare? PMID- 8716065 TI - RIDDOR revisited. PMID- 8716066 TI - [Investigation of the contamination of air and environment in the blood dialysis department]. AB - As the blood dialysis has been applied more widely, the hygenic state of air and furniture surfaces in blood dialysis room and that of dialysis fluid would diretly affect the incidence of nosocomial infections. The results of investigation in three hospital dialysis units showed that: With people walking around, the number of air microbes presented a kenetic variation, ranged from 410 to 1755CFU/m3. That number was positively related to the number of dust particles in the air, which carring certain amount pathogenic of conditional-pathogenic microbes, among which, fungi, Stap. aureus and Pseud. aruginosa acount for 8.38%, 2.99% and 1.39% respectively. Physical of chemical disinfection treatment showed prompt effects (P < 0.01). The thermogenic reaction and infections in dialysis patients were mainly due to microbes containminated in the water for dialysis. Some of these microbes may produce intracellular or and exo-toxin and thermogenic factors. The microbes in dialysis machines could 100% be killed immediately after disinfecting the dialysis tubes with 5% available chlorine. But the germ numer could rise again as the time streched. Our results suggested that certain monitoring and disinfectious procedures must be carried out in hospital critical departments so as to reduce nosocomial infection cases. PMID- 8716067 TI - [Study of infant hair specimens for trace element contents under different conditions of feeding]. AB - The contents of 11 microelements (Cu, Zn, Fe, Mn, Ca, Mg, Al, P, Ba, Se, Sr) in breast milk of 53 postpartum feeding mothers and the hair specimen of 278 healthy infants with different feeding forms (breast feeding, milk and milk mixture) had been measured. The age of the infants was 72 days, the statistical analysis was shown that Zn value of infant hair specimen with breast feeding was significantly higher than the milk feeding (P < 0.01). Breast milk can provide enough Se for the infants. The dispensing milk and the dispensing milk powder were the satisfactory milk feeding substitute in the market now. The improved opinions were suggested. PMID- 8716068 TI - [Emergency treatment and care of hypokalemia paralysis due to gossypol poisoning]. PMID- 8716069 TI - [Promotion of esophagopleural fistula healing by taking food orally]. PMID- 8716070 TI - [Care of lower limb fracture treated with a multifunctional external fixator]. PMID- 8716071 TI - [Nursing care for trans-oral approach operation patients with articulate atlanto epistrophica dislocation during the perioperative period]. PMID- 8716072 TI - [Care of plastic surgery of the palate and pharynx for snoring]. PMID- 8716073 TI - [Open care for mental patients at acute treatment wards]. PMID- 8716074 TI - [Puncture of the Hegu acupoint to accelerate the second stage of labor and to reduce postpartum hemorrhage]. PMID- 8716076 TI - [Establishment of tertiary first class hospital and promotion of standardized control of key department]. PMID- 8716075 TI - [Observation of drainage volume after radical neck dissection]. PMID- 8716077 TI - [Applying and integral system to continuing education for junior nurses]. PMID- 8716079 TI - [An elementary introduction to objective management of the trainees attitude]. PMID- 8716078 TI - [Study of objective-based instruction in basic nursing teaching]. PMID- 8716080 TI - [Care of senior people with psychiatric reactions due to loss of their spouse]. PMID- 8716081 TI - [Care of barium meal treatment for newborn vomiting]. PMID- 8716082 TI - [Nursing care in general medicine]. PMID- 8716085 TI - RNs join first response teams. PMID- 8716086 TI - Nurses key link in the chain of survival. PMID- 8716087 TI - Midwifery in Saskatchewan: working in the community. PMID- 8716088 TI - Nurse abuse. PMID- 8716089 TI - [Message from the Holy Father on the occasion of the 4. World Day of the Patient. February 11, 1996]. PMID- 8716090 TI - [Historical notes on St. John of God. 500th Anniversary 1495-1995]. PMID- 8716091 TI - [Humanization and the quality of life]. PMID- 8716092 TI - [Technical progress in the service of man]. PMID- 8716093 TI - [Professional satisfaction of the nurses at the Lamego district hospital (an exploratory study)]. PMID- 8716094 TI - [Safe handling of antiseptics]. PMID- 8716096 TI - Sexual health care in British Columbia: a model of service delivery. AB - The Sexual Health Program at British Columbia Rehabilitation Society has been providing comprehensive sexual health care to people with spinal cord injuries for the past 20 years. The focus of this paper is to describe this model of service delivery to nurses who may be interested in implementing similar models of service delivery to other health care settings. Sexual health care occurs within the context of the readiness, values and beliefs of the client. The Sexual Health Clinician provides a continuum of consumer-driven services to assist individuals, couples and family members to understand and manage the sexual outcomes of spinal cord injury. As part of the client team, the Sexual Health Clinician provides comprehensive assessment, education, and management for clients and staff. Assessment involves clarifying sexual concerns, establishing present sexual function status using the Sexual Assessment Framework, and exploring past functioning and partnership issues. Concerns are normalized, validated, and support is provided to the client and/or family. This paper will assist nurses to identify the benefits of having a Sexual Health Program as part of spinal cord injury nursing care and be able to identify principles of sexual health care practice that may be able to be incorporated into their present clinical setting. PMID- 8716095 TI - [Nursing procedures in the use of anticoagulant therapy]. PMID- 8716097 TI - The day treatment program: meeting rehabilitation needs for SCI in the changing climate of health care reform. AB - Life expectancy for persons with spinal cord injury (SCI) is somewhat below normal but continues to increase (Devivo & Stover, 1994). Therefore, rehabilitation nurses must develop a way to provide interventions over an entire lifespan for the person with SCI. One setting in which to implement care is the day treatment program, and rehabilitation nurses can be a part of this new care delivery model (Stevens, 1994). Because of increasing regulations with reimbursement and the need for early discharge planning, the Rehabilitation Nursing Department at Spain Rehabilitation Center developed a day treatment program. Patients now have the opportunity to continue their rehabilitation program and become more functional if maximum potential is not reached during their inpatient rehabilitation stay. A nurse serves as the case manager to plan, coordinate, and monitor patient programs and patient care. The expectation of this author is that nurses may facilitate comparable programs in the agencies where they practice. PMID- 8716098 TI - Pain management and SCI--implications for nursing research. PMID- 8716099 TI - Sharing knowledge: an interdisciplinary approach in prevention of urinary tract infection. PMID- 8716100 TI - Cultural context, health and health care decision making. 1994. AB - Decisions about health promotion and illness prevention occur within a cultural context that is influenced by the contemporary context of community and family in addition to the ethnohistorical and language contexts, worldview and sociocultural factors of the particular culture and the available folk and professional health care resources. Using information about the health and care beliefs and values and health care decision making process in negotiating culturally congruent nursing and health care interventions is imperative, especially in a world with limited health care resources and an increasing demand for recognition of cultural diversity. This article uses data on health and health care decision-making from an ethnonursing study of the Old Order Amish to demonstrate the role of cultural context in health care practices and decision making. Leininger's cultural care theory and Hall's conceptualization of high context culture were used to investigate these phenomena. High context features of the Old Order Amish culture are used to explain how Amish are actively involved in decisions and actions taken to promote health and prevent and treat illness using a broad array of folk, alternative and professional services simultaneously. As nurses learn to involve clients in decisions and actions using the guiding principles of cultural care preservation, accommodation and repatterning they will provide culturally congruent care for Amish and other culture-specific groups. PMID- 8716101 TI - Patients', relatives', and nurses' experience of stroke in northern Vietnam. AB - Transcriptions were made of interviews held in Vietnam with five nurses, five people affected by stroke (aged 28-60), and with one relative each. Their stories were analysed as texts by means of a phenomenological hermeneutic method. The role of the nurses in Vietnam appeared to differ from that usually ascribed to nurses in Western institutionalized care. In the former a relative was expected to carry out basic care during the acute stage and to assist socioeconomically thereafter. The interviewees spoke of family bonds in the past, present, and future tenses when narrating their experiences of stroke as nurses, patients, and relatives. This was interpreted as an indication of a consciousness of an essential relatedness. Nurses were aware of playing a temporary and secondary role mainly as assistants: firstly to the doctor by carrying out orders and reporting, secondly to the stroke patient and his or her family by carrying out advanced nursing procedures and giving support. The advantages and disadvantages of involving the family in nursing care can be revealed by obtaining views from a different culture, which can provide a contrast against which constructive criticism of the Western nursing tradition can be made. PMID- 8716102 TI - Massage by nurses in the United States and the People's Republic of China: a comparison. AB - Massage performed by a Chinese nurse is described as observed by the author. A brief history of both Eastern and Western massage is presented. A comparison of theory, indications, methods, and outcomes of massage as performed by nurses from the United States (U.S.) and the People's Republic of China is made. The comparison is based on nursing texts and journals printed in the U.S., the author's personal experience in the People's Republic of China, correspondence with a nurse from the People's Republic of China, and texts and journals printed in English in the People's Republic of China. Massage as a nursing intervention differed on many points of comparison between the two countries. Chinese nurses perform massage over the meridians which are the channels for Chi--the life force of energy. Nurses in the U.S. perform massage over tense muscles. Massage performed by U.S. nurses involves three to five strokes while Chinese nurses use dozens of strokes. In China, massage is considered curative; in the U.S. it is mostly palliative. PMID- 8716103 TI - Leininger's model for discoveries at The Farm and midwifery services to the Amish. AB - This paper is a descriptive report and analysis of a transcultural nurse's experiences immersed in a hippie subculture at The Farm near Summertown, Tennessee. This subcultural group initially was established over 20 years ago as a community with a unique worldview which included pacifistic, vegetarian, and collective values and beliefs. This community prefers health care provided by their own community members who serve as generic care providers and also as folk midwives for home births. Leininger's (1991) Theory of Culture Care Diversity and Universality and her Sunrise Model provided the framework for discovering and understanding this unique subcultural group. The major components of Leininger's Sunrise Model including worldview, cultural values, and lifeways were used in the analysis. The important social structure factors discovered included environmental context, technological factors, religious and philosophical factors, political and legal factors, economic factors, and educational factors. The Farm community's culture care expressions, patterns and practices for health and well being were discovered including generic and folk systems of care. The farm midwives provide primary care and home birthing care to a nearby Old Order Amish community. The Amish culture and health care seeking patterns are discussed including their selective use of generic, folk, and professional care systems. The discoveries that resulted from the application of Leininger's Sunrise Model are presented including implications for transcultural nurse caregiving. PMID- 8716104 TI - A guide to searching on-line for information on transcultural nursing and health. PMID- 8716106 TI - [History of a service project]. PMID- 8716105 TI - Nursing theories and cultures: fit or misfit? PMID- 8716107 TI - [The relationship between nurse and child]. PMID- 8716108 TI - [The nurse-parent relationship]. PMID- 8716109 TI - [Inter-staff relations]. PMID- 8716110 TI - [Pediatrics and pediatric psychiatry]. PMID- 8716111 TI - [The district nurse in National Education]. PMID- 8716112 TI - [The severely asthmatic adolescent patient. Psychopathologic approach]. PMID- 8716113 TI - [Working for effective prevention]. PMID- 8716114 TI - [Volunteers in pediatric service]. PMID- 8716115 TI - [Asthmatic crisis]. PMID- 8716116 TI - [Whooping cough]. PMID- 8716117 TI - Coping with leg ulcers. PMID- 8716118 TI - A better life for the dying. Interview by Anne Manchester. PMID- 8716119 TI - Experimenting in Canterbury. Interview by Anne Manchester. PMID- 8716120 TI - Nursing needs research. Interview by Teresa O'Connor. PMID- 8716121 TI - Changes threaten nurses' role. PMID- 8716122 TI - Growing with the student unit. Interview by Teresa O'Connor. PMID- 8716123 TI - Rebuilding the health system. PMID- 8716124 TI - The challenge of change. PMID- 8716125 TI - 1995 fact sheet: supply of nurses in Florida. PMID- 8716126 TI - Through the eyes of a student. PMID- 8716128 TI - Medication errors: the nursing role. PMID- 8716127 TI - Ethical and legal bases of informed consent in Florida. PMID- 8716129 TI - Leadership in nursing. PMID- 8716130 TI - Yes, you can get there from here--career development (Part III). PMID- 8716131 TI - "A nurse is in charge": Susan Reinhard appointed Deputy Commissioner. PMID- 8716132 TI - Institutional credentialing and priveging for advance practice nurses (APN)--a step by step approach. PMID- 8716134 TI - You aren't getting hired? Better check your employment history. PMID- 8716133 TI - Impairment: today, more than just a matter of substance abuse. PMID- 8716135 TI - Legal issues for nurses. Minors and the ability to consent to HIV testing. PMID- 8716136 TI - RN examines the "death of public health". PMID- 8716137 TI - Teleconference on HIV and pregnancy set for June 18. PMID- 8716138 TI - What's in an hour? PMID- 8716140 TI - Career clinic. Strategies for becoming a consultant in your field. PMID- 8716139 TI - PNA member offers dramatic portrayal of Margaret Sanger--nurse, social reformer. PMID- 8716141 TI - Parental attitudes, knowledge, and behaviors and childhood injury in North Dakota. PMID- 8716143 TI - A JCAHO primer on the work of restructuring. PMID- 8716142 TI - An interview with Rep. Bill Oban. Interview by Bonnie Staiger. PMID- 8716144 TI - Interfacing labor laws with decision-making structures. PMID- 8716145 TI - Do we get what we deserve or what we create? PMID- 8716146 TI - ANA applauds NLRB ruling on RN status: ruling means RNs can advocate for patients without fear of firing. PMID- 8716147 TI - Union is not a dirty word. PMID- 8716148 TI - Clinical incident where I made a difference. PMID- 8716149 TI - Through the eyes of a student. PMID- 8716151 TI - Yes, you can get there from here--career development (Part II). PMID- 8716150 TI - Nurse attorney notes. PMID- 8716152 TI - The perception of self esteem in female victims of sexual assault. PMID- 8716154 TI - A rural nursing experience. AB - This paper describes student observations and responses to an introduction of nursing in a rural area. As many rural areas of the United States continue to face a shortage of nurses, working in a rural area is an option for employment upon graduation. Independence is a major factor in choosing a rural setting for a nurse anesthesia practice. PMID- 8716153 TI - Utilization of nursing research by public health nurses. PMID- 8716156 TI - Community-based nursing case management: from vision to reality. PMID- 8716155 TI - Use of nursing research in practice? Keep reading! PMID- 8716157 TI - Kansas State Nurses Association: violence assessment in hospitals project general summary and comparison. PMID- 8716158 TI - Celebrate Nurses' Week, reflect on personal responsibility to adapt. PMID- 8716159 TI - Program provides behind-scenes benefits to keep skills up to date. PMID- 8716160 TI - Nurses can do it all. PMID- 8716161 TI - Career clinic. Determining the causes of an unsuccessful job search. PMID- 8716162 TI - Behaviors, attitudes, and knowledge related to drug and alcohol prevention curricula in North Dakota seventh through twelfth grade students. PMID- 8716163 TI - 10 tips for a top interview: use these proven techniques to land the job you want. PMID- 8716164 TI - Initiating ethics consultations. PMID- 8716165 TI - Nursing unions: are they necessary? PMID- 8716166 TI - Transformational nursing. PMID- 8716167 TI - Look before you leap. PMID- 8716168 TI - Change in health care organizations: an overview. PMID- 8716169 TI - ANA position statement on restructuring, work redesign, and the job and career security of registered nurses. PMID- 8716170 TI - Critical paths: a tool to improve skiing on the slopes of managed care. PMID- 8716171 TI - Partnerships for primary care in a changing health care system: a Tennessee nursing model. PMID- 8716172 TI - To smoke or not to smoke--this is a question. To age or not to age is not a question but whether to age actively and involved is a question. PMID- 8716173 TI - Career clinic. Human relations skills--critical factors in career success. PMID- 8716174 TI - Unemployment law and the registered nurse: when do you have to accept a job? PMID- 8716175 TI - Methods for nurses to achieve empowerment. PMID- 8716176 TI - Is the enemy among us? It's my opinion. PMID- 8716177 TI - Expertise in caring. PMID- 8716178 TI - From the Ohio Board of Nursing: continuing nursing education. PMID- 8716179 TI - Taking the fear out of AIDS nursing: voices from the field. AB - This paper discusses the findings of a quantitative study of nurses' willingness to care for patients with AIDS. It identifies several factors that enable nurses to confront the fear of AIDS nursing and to turn fear into compassion, and negative attitudes into positive ones. These factors include professional values, organizational support, group identity, patients' responses, spirituality, family and friends with AIDS, knowledge about AIDS, and the importance of personal choice. The value of these factors are further supported through personal statements made by nurses on AIDS-dedicated units as they express their feelings, thoughts, and positive experiences and expectations regarding AIDS nursing. PMID- 8716180 TI - Group process among novice students. AB - This paper explores the experiences of a group of registered nurses returning to graduate school. The organizing framework used is an integration of Tuckman and Jensen's group process theory and Benner's novice to expert model. PMID- 8716181 TI - Concept mapping and synthesizers: instructional strategies for encoding and recalling. AB - The process of knowledge acquisition requires that nursing students obtain concepts which must be encoded, stored, and later recalled from long-term memory. To assist students with these processes, teaching strategies that structure content should be used throughout the curriculum. This article describes how the instructional strategies of concept mapping and synthesizers were used to facilitate nursing students' encoding and recalling of pharmacology concepts in an undergraduate pharmacology course. PMID- 8716182 TI - Nursing sites on the World Wide Web. PMID- 8716184 TI - HIV survival rates. PMID- 8716183 TI - Professional development. Dementia: knowledge for practice (continuing education credit). PMID- 8716185 TI - Adverse reactions. PMID- 8716186 TI - They talk, but do they communicate? PMID- 8716187 TI - Fans told to play the health card. PMID- 8716189 TI - Multiple sclerosis. A case of mind over matter? PMID- 8716188 TI - Multiple sclerosis. Meeting multiple needs. PMID- 8716190 TI - Multiple sclerosis. Living with the enemy. PMID- 8716191 TI - Multiple sclerosis. An exercise in hope. PMID- 8716192 TI - Planning and carrying out a clinical study: the first steps. AB - This paper is the first of two explaining the methods which may be used in clinical evaluation. It describes how a project or study is planned to meet its aims. It covers issues related to the sample, interventions and the measures employed to evaluate the interventions. PMID- 8716193 TI - Systems of life. Male sexual function. PMID- 8716194 TI - Hand-washing. Can ward-based learning improve infection control? AB - Post-registration education and practice (PREP) requires nurses to become responsible for their own professional updating. For those in clinical posts this may mean obtaining study leave. This paper argues that there is a real need for the NHS to look at new ways of allowing nurses to study and makes the case for ward-based learning. PMID- 8716195 TI - How new nurses manage work and family responsibilities. AB - The predominantly female nursing workforce means that issues such as combining work and child care responsibilities are central to people's careers and to workforce planning. A questionnaire survey of newly qualified registered general nurses provided valuable information about the circumstances, experiences and plans of 936 women. Implications for the availability of flexible working, career guidance and opportunities for professional development are discussed. PMID- 8716196 TI - Spasmodic detection. PMID- 8716198 TI - Welcome complaints. PMID- 8716197 TI - Stoma care: patient support. PMID- 8716199 TI - Continence. Charting new waters. PMID- 8716200 TI - Continence. Behind closed doors. PMID- 8716201 TI - The public health policies of the EU must continue to include cancer. PMID- 8716202 TI - Who has the power? Some problems and issues affecting the nursing care of dying patients. AB - Nurses play a central role in the care of dying people, yet they may find themselves marginalized, devalued or in conflict with other health professionals. This paper examines a number of problems and issues related to nursing power and control in the workplace. The factors we consider are: the relative effects of personal and structural influences; the 'medicalization' of dying; the demands and challenges of interdisciplinary work; ethical issues raised by euthanasia and AIDS; patients' rights to make informed decisions about their care; philosophies and policies of health care. PMID- 8716203 TI - Patient advocacy: a role for nurses? AB - There is much controversy surrounding the issue of nurses in the role of patient advocate. Cancer patients are vulnerable due to their diagnoses, the anxiety that it generates, the nature of the disease and subsequent treatment and may diminished autonomy as a result. It is questionable whether the nurse is able to promote the best interest of the patient in this situation. Models of advocacy and the nurse-patient relationship are discussed. The main arguments for taking on the role of patient advocate include: reference to long periods of time spent with patient and the chance to build a relationship; on duty to care and the duty to do no harm. Criticisms of the appropriateness of advocacy role for nurses include: inability for the patient to choose his or her nurse; promotion of self interests; charges of paternalism; challenges from other health care professionals and multiple and/or conflicting obligations. It is concluded that perhaps the most ethical way of caring is by empowering patients and promoting self-advocacy. This concept needs to be further explored. PMID- 8716204 TI - Ethical and practical problems of early anti-cancer drug trials: a review of the literature. AB - Early clinical trials for new anti-cancer drug treatments typically use patients with cancer as research subjects. This paper identifies some of the ethical and practical concerns that arise from the recruitment of a vulnerable group of patients and their exposure to a drug of unknown risk or benefit. This review discusses the ethical principles related to recruitment and informed consent in cancer trials, and indicates that there is a lack of consensus concerning the requirements, process and practice of informed consent. It is suggested that, as yet, little is known about patients' decision making framework in this situation, and the need for further work that concentrates on the patient's point of view is highlighted. The paper concludes by discussing some of the difficulties associated with obtaining patients' opinions, and suggests that the use of a qualitative approach may overcome some of these problems. PMID- 8716205 TI - Statutory and voluntary sector palliative care in the community setting: National Health Service professionals' perceptions of the interface. AB - Following the rapid service development brought about by the hospice movement, specialist palliative care services are involved with up to 50% of all patients dying with cancer in the United Kingdom, although the primary health care team remains the main provider of community based palliative care. This paper discusses findings from a survey of palliative care provision in the south west of England, and describes the perceptions of the primary care team (general practitioners and district nurses) about the interface between themselves and voluntary sector specialist palliative care providers (hospice in-patient units, hospice home care nurses and other charitably funded specialist palliative care nurses). The voluntary sector services are run with a mixture of funding from charitable sources (public donations, legacies, charitable trust moneys), and statutory funding (grants and recurrent contracts from central government, district health commissions, and local health care trust). The interview and questionnaire data suggest that the voluntary sector services are perceived variably as substituting, supplementing, complementing and duplicating the services provided by the primary care team. Drawing attention to these dimensions and the ambivalence sometimes felt by general practitioners and district nurses could provide a means of negotiating consensus on appropriate professional tasks and facilitating interprofessional practice in what is increasingly a mixed economy of statutory and voluntary funded health care. PMID- 8716206 TI - What do cancer patients identify as supportive and unsupportive behaviour of nurses? A pilot study. AB - Social support has been claimed to have positive effects on a variety of outcomes, including physical health, mental well-being and social functioning, and yet, its nature, meaning and measurement are still being debated in the literature. This pilot study set out to identify, within the theoretical framework of the social support literature, supportive and unsupportive nurse behaviours as perceived of by eight hospitalized patients diagnosed with a haematological malignancy. The findings of the semi-structured interviews appear to suggest that emotionally supportive behavior patterns are the most frequently identified helpful nurse interactions reported by individuals with cancer, followed by informationally supportive behavior. The most frequently identified unsupportive nurse behaviours were those perceived of as being devoid of an emotional component. Tentative conclusions for practice are drawn from these findings and considerations for future research are presented. In accordance with work by Dunkel-Schetter (1984) the terms supportive and unsupportive are operationalized throughout this paper as the more familiar terms helpful and unhelpful. PMID- 8716207 TI - Fatigue in cancer: a phenomenological perspective. AB - Fatigue is a frequently encountered symptom in cancer populations. This study aimed to describe the experience of fatigue from the perspective of cancer patients who had recently completed a course of chemotherapy. A phenomenological method was used. The themes which emerged from the data indicated both a shared and individual experience of fatigue. These incorporated: the nature of fatigue; the causes, consequences, strategies for coping with fatigue; and the trajectory of the fatigue experience. Issues arising from the nature of phenomenological inquiry and the research were also elicited from this study. An understanding of the fatigue experience for this population and the use of phenomenology have implications for the nursing profession's knowledge base and for clinical practice. PMID- 8716208 TI - Palliative care needs in a district general hospital: a survey of patients with cancer. AB - Although sophisticated treatment of cancer requires the resources of specialist cancer treatment centres, most patients with cancer still undergo initial diagnostic investigation and treatment in district general hospitals (DGHs). The DGH frequently remains the principle site of referral for management of symptoms and terminal care. This survey was carried out at a DGH which has no palliative care services. It aimed to demonstrate the need for such services by collecting data on inpatients with cancer and interviewing members of the nursing staff. Of 63 patients studied, 76% were admitted as emergencies, 50% had newly diagnosed cancer and 27% died in hospital. The majority (85%) were cared for on general medical, surgical and care of the elderly wards. Assessment of patients' symptoms suggested that 39 (62%) might have benefited from the services of a palliative care team. Interviews with nursing staff highlighted the need for improved communication between professionals, increased staff education and support, and highlighted the particular difficulties that exist in caring for patients with advanced cancer and their families on busy acute general hospital wards. Imaginative and flexible approaches to the design and delivery of palliative services are essential if patients with cancer in DGHs are to receive the highest standards of care at all stages of their illness. PMID- 8716209 TI - The effects of platinum chemotherapy on essential trace elements. AB - The effects of cisplatin chemotherapy on the metabolism of essential trace elements were investigated in 12 patients before and after treatment with cisplatin. In serum, the mean post-treatment concentrations of Cu 913.91 mumol 1 1), Zn (9.57 mumol 1-1) and Mg (0.54 mumol 1-1) were significantly reduced compared with the pre-treatment levels 919.35, 11.86 and 0.67) while Se, caeruloplasmin and C-reactive protein concentrations were unaltered. Urinary excretion of Cu, Mg and Zn were enhanced. The urinary N-acetyl-beta-D glucosaminidase activity (a marker of proximal renal tubular dysfunction) was also increased and suggests that the mechanism for decrease of certain trace elements in serum during treatment could be increased urinary excretion caused by impaired cellular metabolism. It is not clear whether the loss of trace elements via the urine has any implication for the clinical status of cancer patients treated with cisplatin. PMID- 8716210 TI - Board of Nursing holds hearings on unlicensed care. PMID- 8716211 TI - Teaching and learning through mentorship. PMID- 8716212 TI - Becoming a nurse entrepreneur. PMID- 8716213 TI - Intrapreneurship: bloom where you're planted. PMID- 8716214 TI - Nursing in the new millennium. PMID- 8716215 TI - Nurse practitioner research network: patterns of practice in Northeast Tennessee. PMID- 8716216 TI - Reflections on a 58-year career. Interview by Carol McShane. PMID- 8716217 TI - Nurses must tell nursing stories. Interview by Carol McShane. PMID- 8716218 TI - So, how important is knowing your rights? PMID- 8716219 TI - Case study: a personal look at health services in Bolivia. Omaha nurse practitioner experiences "dropped chin syndrome". PMID- 8716220 TI - Career corner. Creating an image, preparing your resume. PMID- 8716221 TI - Politically speaking: an interview with Representative Pepper. PMID- 8716222 TI - Restructuring: getting the big picture. PMID- 8716223 TI - Continuing education: should it be mandatory? PMID- 8716224 TI - To address the world from a political perspective. PMID- 8716225 TI - Leading the race. PMID- 8716226 TI - The pursuit of politics. PMID- 8716227 TI - The value of fair grading. PMID- 8716228 TI - Proceed with political care. PMID- 8716231 TI - Leadership and nursing: Part Three: Traditional attitudes and socialisation. PMID- 8716230 TI - Commissioning health: a fresh start for Wales. PMID- 8716229 TI - Towards a primary care-led NHS. PMID- 8716232 TI - Contracts for good morale. PMID- 8716233 TI - Doing the right thing? PMID- 8716234 TI - Incorporating cultural sensitivity into educational programs is an important consideration for nurses. PMID- 8716235 TI - Midwives in current debate and in the future. PMID- 8716236 TI - An unplanned birth: an appeal. PMID- 8716237 TI - Where there are no midwives. AB - Nepal is a country with immense health problems affecting women and their newborn infants. As a lecturer at the Nursing Campus, Maharjgunj, Kathmandu, I have been assisting with the midwifery education of student nurses. Student nurses in the second year of a three year certificate in nursing program are educated to care for antenatal women, conduct normal deliveries and provide postnatal care. On completion of their studies, the graduates are able to be employed in maternity hospitals and village health posts. This article outlines my experiences as a lecturer and clinical educator in Nepal, a country where there is no specialised educational program available in midwifery. PMID- 8716238 TI - Criteria for successful breastfeeding: mothers' perceptions. AB - A descriptive study was conducted to determine the criteria Western Australian mothers use to decide whether their breastfeeding was successful. Successful breastfeeding tends to be measured by duration with minimal research examining breastfeeding success from mothers' perspectives. Telephone interviews were conducted with 183 mothers who were asked to classify their breastfeeding as successful or unsuccessful and identify criteria used as a basis for their decision. Nineteen women (10 unsuccessful, 9 successful) were then interviewed in depth. Content analysis revealed breastfeeding success or failure to be a personally defined experience. Four categories of criteria used to determine breastfeeding success emerged: giving, persistence, meeting expectations and accomplishing personal goals. Nurses play a significant role in promoting breastfeeding. Therefore, encouraging understanding of the mother's perspective values the uniqueness of each experience and allows nurses to assess, acknowledge and reinforce a mother's perceptions of success. PMID- 8716239 TI - Dying for the cause. PMID- 8716240 TI - [Evolution of blood transfusion and monitoring]. PMID- 8716241 TI - [Facilities for blood transfusion. Their role and functioning]. PMID- 8716242 TI - [Blood transfusion and monitoring at the community hospices of Lyon]. PMID- 8716243 TI - [Blood transfusion and monitoring. Blood products for transfusion. Indications]. PMID- 8716244 TI - [Blood transfusion and monitoring. Immuno-hematologic data]. PMID- 8716245 TI - [Blood transfusion and monitoring. Immediate transfusion reactions. Erythrocyte transfusion]. PMID- 8716246 TI - [Blood transfusion and monitoring. Post-transfusion hemochromatosis]. PMID- 8716247 TI - [Blood transfusion and monitoring in the hospital. Logistical aspects]. PMID- 8716248 TI - [Immunological reactions connected with transfusion. Prevention]. PMID- 8716249 TI - [Transfusion check list]. PMID- 8716250 TI - [Organ donation. Study of 303 families]. PMID- 8716251 TI - [Forty years. Contribution and dynamics of the journal Soins]. PMID- 8716252 TI - [Forty years. Nursing care, AIDS and jurisprudence]. PMID- 8716253 TI - [Plastic materials. Their part in the quality of care and in drug administration]. PMID- 8716255 TI - [Anticholinergic anti-Parkinson agents]. PMID- 8716256 TI - [Very young autonomy]. PMID- 8716257 TI - [Hygiene in intensive medicine--2]. PMID- 8716258 TI - [Experiences of an Austrian nurse in Germany]. PMID- 8716259 TI - [Doing research is trying out]. PMID- 8716261 TI - [Nursing and care of diabetic children and adolescents]. PMID- 8716260 TI - [Diabetes--definition and classification]. PMID- 8716262 TI - [Discharge of a patient with diabetes mellitus]. PMID- 8716263 TI - [Immediate measures in diabetic accidents]. PMID- 8716264 TI - [Our child will be a nurse. A study shows that three quarters of all parents would support this career choice]. PMID- 8716265 TI - [Nursing research as a contribution to good health]. PMID- 8716266 TI - [Diabetes training: the Burgenland model]. PMID- 8716267 TI - The goodness of measurement. PMID- 8716268 TI - Pressure ulcers: a preventable American tragedy. PMID- 8716269 TI - Planning for health care and policy changes. PMID- 8716270 TI - 31P NMR spectroscopic analysis of wound healing: the effect of hydrocolloid therapy. AB - With the advent of managed care, wound care professionals have limited time to heal chronic wounds. They need to know whether the repair process is progressing or stagnating in response to treatments. Phosphorus-31 (31P) nuclear magnetic resonance (NMR) spectroscopic measurements of chronic wound biochemistry yields rapid knowledge of whether a wound is generating, storing, or using energy. We used 31P NMR analysis on biopsy samples to explore the energy status of two chronic non-healing leg ulcers, before and after the first week of treatment with two low-pH hydrocolloid materials. Energy generation (i.e., energy "charge") was initially low in both wounds and was significantly elevated after 1 week of treatment. Earlier work has shown that leg ulcer pathophysiology is altered during the first week of hydrocolloid treatment. This work traces the origins of such effects deeper into the cellular biochemistry and correlates the measures with the final healing outcome. 31P NMR spectroscopy may provide a real-time biochemical "fingerprint" that shows clinicians the healing status of a questionable wound. Further study is needed to confirm the reliability and validity of 31P NMR spectroscopy as a predictor of healing outcomes in other wound environments. PMID- 8716271 TI - Changes in occipital capillary perfusion pressures during coronary artery bypass graft surgery. AB - This study compared tissue interface pressures (TIPs) on the occiput of 25 volunteer patients undergoing coronary artery bypass graft (CABG) surgery. Pressures were measured using the Gaymar pressure gauge and electropneumatic sensor before and after induction of anesthesia, after the patient was placed on the cardiopulmonary bypass pump, every 30 minutes throughout the procedure, immediately after the patient came off the pump, and before the incision was closed. Significant differences were found over time on mean TIPs and mean arterial pressures (MAPs). Pre-induction mean TIP was significantly higher than TIPs recorded on-pump, post-pump, and prior to closure. The highest average MAPs were at pre-induction; these were significantly higher than MAPs recorded at all other times. The TIPs recorded during surgery were high enough to put the patient at risk for circulation impairment and pressure ulcer development. Therefore, we recommend repositioning patients' heads regularly during surgery and assessing skin postoperatively for alopecia and pressure ulcers. PMID- 8716272 TI - Comparison of 90 degrees and 30 degrees laterally inclined positions in the prevention of pressure ulcers using transcutaneous oxygen and carbon dioxide pressures. AB - Although proper positioning is an important measure in pressure ulcer prevention, few objective measurements have been reported of the effects of different positions on oxygen supply to the skin area in direct contact with the mattress. This study compared the effects of 30 degree and 90 degree laterally inclined positions on the cutaneous oxygen supply to the skin through the measurement of transcutaneous oxygen pressure (TCPO2) and transcutaneous carbon dioxide pressure (TCPCO2) on the trochanteric (T) and the retro-trochanteric (R) areas in 20 healthy volunteers. Resting values were TCPO2 (T) 69.4 mm Hg (SD = 10.4), TCPCO2 (T) 36.3 mm Hg (SD = 5.1), and TCPO2 (R) 69.9 mm Hg (SD = 9.2). In the 90 degree laterally inclined position, the TCPO2 (R) value (68.2 mm Hg [SD = 9.9]) did not differ significantly from the resting value. These results show a dramatic impairment of oxygen supply to the skin in the 90 degree laterally inclined position but not in the 30 degree laterally inclined position. Additional studies of patients immobilized for long periods are needed to confirm the best positioning to prevent pressure ulcers. PMID- 8716273 TI - Increased energy needs in patients with quadriplegia and pressure ulcers. AB - Health individuals with quadriplegia generally have a reduced metabolic rate. However, individuals with quadriplegia who develop pressure ulcers may have an elevated metabolic rate. In this study, energy expenditure in 16 individuals with quadriplegia and pressure ulcers (PU-QUAD) was compared to the energy expenditure in 16 individuals with quadriplegia but no pressure ulcers (NPU-QUAD) and 16 healthy non-spinal cord injured subjects (controls). Resting energy expenditure (REE) was measured by indirect calorimetry. Both measured REE (t(30) = 2.38, p = 0.24) and percent predicted REE (t(30) = 3.23, p = .003) were significantly higher in subjects with quadriplegia and pressure ulcers compared with subjects with quadriplegia but no pressure ulcers. On average, REE in the PU-QUAD subjects was nearly equal to the absolute energy expenditure of healthy non-spinal cord injured controls. To ensure optimal care of patients with quadriplegia and pressure ulcers, quantification of energy expenditure with provision of adequate caloric intake is recommended. PMID- 8716274 TI - Implementing patient-focused care: an eight week fast-track approach. PMID- 8716275 TI - Training nursing assistants for expanded roles as PCT's. PMID- 8716276 TI - Albendazole may reduce diarrhoea in HIV patients. PMID- 8716277 TI - Waste away. PMID- 8716278 TI - Epilepsy care using inside information. Interview by Catharine Sadler. PMID- 8716279 TI - Confined to barracks? PMID- 8716280 TI - Bowel management and nursing's hidden work. AB - This paper looks at the nature of nurses' role in bowel management. It discusses how this fundamental and important work is hidden from public view and is therefore undervalued and ignored. The reasons for this are highlighted. The author calls for nurses to publicise their continuous and substantial achievements in this area. PMID- 8716281 TI - Assessing constipation. AB - This paper discusses the generic view of defaecation and examines the mechanisms contributing to constipation. Although it is recognised that there are specific causes that will require medical intervention, for the purpose of this paper only the general clinical management will be addressed. PMID- 8716282 TI - Movement through change. AB - Constipation is often overlooked when it comes to assessing clients' needs. This paper, by a team caring for a group of clients with severe learning disabilities, looks at how constipation can be managed and calls for changes to be made in the care of patients experiencing this difficulty. PMID- 8716283 TI - Clients express preference for one-stop sexual health shop. AB - The concept of sexual health is becoming more prominent. However, health services relating to the maintenance of sexual health do not necessarily use the available resources to their best advantage. This paper examines developments in the area of sexual health and presents the results of a survey carried out in Eastbourne and Ipswich, which indicates that attenders of various clinics dealing with sexual health would welcome services such as family planning and genitourinary medicine being combined on one site. PMID- 8716284 TI - Youngsters get an introduction to sexual health clinics. AB - This paper describes the development of mock sexual health clinics for young people in an area with high rates of teenage pregnancy and poor awareness of sexual health services. The multi-disciplinary initiative was developed in partnership with local schools and aimed to promote access to sexual health and contraceptive services by breaking down barriers and misconceptions. Subsequent registrations of young people at local family planning clinics have increased dramatically and a full evaluation is being undertaken. PMID- 8716285 TI - Please believe me. PMID- 8716286 TI - Diversion tactics. PMID- 8716287 TI - Nursing abroad. Care against all the odds. PMID- 8716288 TI - Wheelchairs: what the NHS can offer. PMID- 8716289 TI - The death of ideals. PMID- 8716290 TI - Mental health. Partners, not adversaries. PMID- 8716291 TI - Mental health. General reflections. PMID- 8716292 TI - Mental health. Positive force for change. PMID- 8716293 TI - Scope for change. PMID- 8716294 TI - President's message: on paying now or later, and asking those who know. PMID- 8716295 TI - On the dark side of 'corporate care': conflicts of interest and the growing need for regulation. PMID- 8716296 TI - Another case of gamma hydroxybutyrate (GHB) overdose. PMID- 8716297 TI - A case of ciguatera fish poisoning. PMID- 8716298 TI - A study of factors influencing ED patients' length of stay at one community hospital. AB - OBJECTIVE In 1993 the administration of Kennewick General Hospital selected a facilitator to assist an ED task force in isolating and testing aspects of operations for quality improvement. The primary problem selected was reducing the length of stay (LOS) for patients. The department had an annual census in excess of 33,000, with seven rooms and 96 hours of registered nurse staffing per day. METHODS The task force selected 11 factors considered to possibly reduce LOS. The Taguchi method of robust quality improvement, an engineering quality improvement method, was used to determine the best average impact for each factor. Testing was conducted for an 11-day period. RESULTS The top three factors with significant impact on decreasing patient LOS were adding an additional physician, an additional laboratory professional, and implementation of a policy that allowed the emergency physician to hold a patient for possible admission in an inpatient unit while attempting to locate and communicate with the patient's private physician. DISCUSSION The study results met some expectations but also provided some unexpected findings for the ED staff. Some factors were implemented to reduce patient LOS and others for a different improvement value to patient care (i.e., a private triage room). Some highly effective factors were not implemented because of significant changes in department volume. PMID- 8716299 TI - A comparison of two noninvasive methods of blood pressure measurement in the triage area. AB - OBJECTIVE In triage of patients in the emergency department, nurses can identify conditions such as elevated blood pressure that can lead to serious health problems. It is imperative that the method used to evaluate blood pressure is dependable and accurate. The introduction of new technology may yield results that are not comparable to traditional measures. Therefore we compared the traditional auscultatory method of measuring blood pressure with the use of an automated device. METHODS The blood pressure of 100 healthy adult volunteers was measured first with the traditional cuff method and then with an automated device (IVAC model 4200). The measurements were recorded and then compared for each individual. RESULTS The mean difference between the paired systolic measurements was 4.3 mm Hg (p = 0.0001). The mean difference between the paired diastolic measurements was 1.3 mm Hg. There was a clinically significant difference (> 9 mm Hg) in 37% of the systolic readings and 26% of the diastolic readings. In 76% of those clinically significant systolic differences, the IVAC 4200 measurement exceeded the manual method. For the clinically significant diastolic differences, 58% of the IVAC 4200 measurements were greater than the manual method. CONCLUSION Clinicians must be cautious when using new technology to measure blood pressure. Results may not be comparable with results obtained with the traditional method. PMID- 8716300 TI - Competency verification for conscious sedation. PMID- 8716301 TI - Neonates and premature infants: overview of differences and ED management. PMID- 8716302 TI - Appropriate use of air bags: a word of caution. PMID- 8716303 TI - Reversing life-threatening cardiac glycoside intoxication with digoxin immune Fab. PMID- 8716304 TI - The mobile emergency department: one nurse's foresight into the future. PMID- 8716305 TI - Air bags and lower extremity trauma: implications for flight crew members. PMID- 8716306 TI - Bar coding technology and its applications in emergency care. PMID- 8716307 TI - Nurse-physician communication breakdown: is it a basis for nurse liability? PMID- 8716308 TI - I have heard that nurse practitioners are cost-effective providers of ambulatory care. PMID- 8716310 TI - Nursing research: our first time. PMID- 8716309 TI - Two important lessons: caution with telephone triage and believing the caregiver. PMID- 8716311 TI - Sexual assault: clinical issues. VOICES. PMID- 8716312 TI - Vaginal laceration caused by water skiing. PMID- 8716314 TI - Deployment to Haiti: an emergency nurse's story. PMID- 8716313 TI - A 52-year-old man with wheezing, fever, and abdominal pain. PMID- 8716316 TI - The difference in both our lives. PMID- 8716315 TI - An experience shared. PMID- 8716317 TI - Motor vehicle injury prevention grant projects under way. PMID- 8716318 TI - A look at our new emergency department: New England Medical Center, Boston, Massachusetts. PMID- 8716319 TI - The effect of intravenous fluid infusion on blood and urine parameters of hydration and on state of consciousness in terminal cancer patients. PMID- 8716320 TI - What is wrong with this picture? PMID- 8716321 TI - Elsa's door. PMID- 8716322 TI - Utilization review of hospice home care. PMID- 8716323 TI - Storytelling for grieving children. PMID- 8716324 TI - Training volunteers and professionals to work with grieving children and their families. AB - This article presents five foundational principles important for the training of professionals and volunteers who work with grieving children and their families. The principles discussed are: first, that expressing feelings is part of the healing process and requires safety and trust; second, that grieving is a process unique to each person; third, that working with grieving children and their families touches our own grief; fourth, that process is more important than content; and fifth, that working with grieving children and adults is a three fold commitment. The importance of training and concluding remarks concerning inspiration are also included. PMID- 8716325 TI - Recognizing delirium in terminal patients. PMID- 8716326 TI - A comparison of hospice programs based on Medicare certification status. AB - This article presents results from a study of 119 hospice programs in the United States. Personal interviews and questionnaires were utilized to collect data about hospice programs, their directors, nurses, social workers, and chaplains. Specifically, this article describes reasons programs sought Medicare certification, and the perceived advantages and disadvantages to being a Medicare certified program. Characteristics of both certified and non-certified programs are presented, and examined for differences. Potential access barriers such as restrictive admission criteria are examined in this article. Finally, perceptions of staff about hospice services in certified and non-certified programs are compared. Results from this study indicate that Medicare certified programs have longer lengths of stays, were more likely to include a nurse on the first visit, and billed patients more frequently than noncertified programs. Volunteer use was lower in the Medicare certified programs. Staff in Medicare certified programs were much more likely to view patients' medical needs as the primary focus of their programs. Results from this study suggest that Medicare certified programs may reflect a more medical model of palliative care. PMID- 8716327 TI - Pain, music creativity and music therapy in palliative care. AB - An analysis of the music therapy literature yields numerous reports to support the role of music in the alleviation of pain in palliative care. Four theoretical perspectives that support why many patients report reduced pain sensation after music therapy include: the psychological relationship between music and pain; the psychophysiological theory; spinal mechanisms involved in pain modulation; and the role of endorphins. Considerations significant to the use of music in pain relief include how music, used inappropriately, can aggravate pain sensation. Case studies, which include the use of creative music therapy techniques, point to the efficacy of music therapy in alleviating the pain experiences of both palliative care patients and their significant others. PMID- 8716328 TI - Massage therapy: an added dimension in terminal care. PMID- 8716329 TI - The Sacred Circle: a conceptual framework for spiritual care in hospice. AB - Hospice care has consistently recognized the need to integrate spiritual care into holistic plans of care for dying patients and their families. Designing and implementing spiritual care interventions can be potentially difficult for hospice practitioners who have not had specific training in theology or pastoral care. Matthew Fox, a theologian, has developed a model of spiritual development that utilizes an ecumenical, ethical framework that can be directly applied to the care of hospice patients and families. This model employs a Sacred Circle approach that begins with an emphasis upon the sense of awe and wonder (the Via Positiva), moves into the next cycle by recognizing problems and negative emotions (the Via Negativa), that then flows into the creative solutions to problems (the Via Creativa), which finally transforms the problem into a new level of understanding (the Via Transformativa). PMID- 8716330 TI - The importance of preventing medication errors. PMID- 8716331 TI - Human factors design--what is it and how can it affect you? AB - Human factors play a significant role in the design and production of medical devices, appealing to the user as easy to use as well as cost-effective. Incorporating human factors into the design process takes strategic planning and clinical observations to accurately determine the most safe and efficient delivery of care. This article explains the process in developing human factors that enhance product design and how they ultimately benefit the patient. PMID- 8716332 TI - Intravenous medication errors. AB - In a clinical study, it was reported that based on the number of adult patients admitted to two hospitals, medication errors amounted to almost 4,000, an alarming figure considering that many were thought to be preventable. Today's healthcare environment dictates the type of care patients receive and from whom, and the potential for medication errors. This article discusses the factors concerning the prescribing of i.v. medications, preparing the medications, and administering them competently by nurses and pharmacists. PMID- 8716333 TI - Legal implications/risk management. AB - The safe delivery of medication, regardless of how skilled the clinician, involves some risk and liability. To minimize these risks, nurses and i.v. professionals must practice prevention techniques. Risk management in today's healthcare environment involves implementing education and awareness programs as well as carefully identifying the problems and instituting solutions to positively affect all those involved. This article identifies the risks and legal implications involved in patient care and addresses how to promote positive patient outcomes. PMID- 8716334 TI - Continuum of care and human factors design issues. AB - Managed care now affects every facet of healthcare. Legislation and reform play a significant role in the reduction of nursing staffs, whereas added responsibilities fall on the shoulders of those who remain. As trends shift, so does the quality of care, along with the practice setting. This article focuses on managed care systems and the role of human factors engineering in the continuum of care. PMID- 8716335 TI - Implementation of physician orders for desired patient outcomes. AB - Human factors engineering was incorporated into the design of medical devices to reduce human errors and enhance the quality of patient care. This study will discuss medication delivery through the use of the IVAC Signature Edition infusion pump and its capability to deliver safe, efficient intravenous therapy. PMID- 8716336 TI - [Divergences and agreements between the nursing model for diabetic patients and Orem's self-care deficit theory]. AB - The divergences and convergences between the Nursing Assistance model for diabetic patients and the OREM's Self-care Deficit theory are presented. The model studied may be considered an educational program for diabetic patients, differing on the conceptual aspects of OREM's theory. PMID- 8716337 TI - [Care of children with cancer: analysis of the work process]. AB - This study tries to analyse some principle of the working process in the assistance of children with cancer, based on the Model of the Technological Organization of Work. It started from a theoretical principle that nursing is a social practice and so is historically structured, socially articulated, and so nursing is part of the collective process of work. We pointed out toward the fact that the process of the nursing work in assisting a hospitalized child, that it is under a transformation phase, going through the exclusively clinic model to another one, which has some characteristics of amplification to the theoretical reference and a new instrumentalization. PMID- 8716339 TI - [The diagnostic process and how to teach it]. AB - The subject of the present study is the teaching of the ability of diagnosing to nursing students. It has the general objective of verifying the adequacy of the elaboration of the analysis and synthesis process and its relation to the establishment of nursing diagnosis. After the development of the programmatic content of the Medical Nursing Discipline, students individually elaborated, facing a hypothetic situation, the diagnosis process (RISNER, 1986), obtaining a total of 106 diagnosis formulations (Taxonomy I of North American Nursing Diagnosis Association). The major difficulties were in the areas of "the establishment of relations and "data clustering". Authors suggest the stimulation of the development of the process of thinking inherent to analysis and synthesis since the beginning of undergraduate course. PMID- 8716338 TI - [Nursing research and positivism]. AB - The authors discuss positivism foundations; the most remarkable influences to comtian's philosophy structuration; how Comte's ideas, as doctrine in so far as method, dominate society's thoughts, passing beyond XIX century and reaching XX century; and how the positivist doctrine and empirical method influenced Nursing conceptions about science and about human being/environment/disease. PMID- 8716340 TI - [The politics of gender and of women's health in Latin America: Peru. 1]. AB - This study includes a presentation and analysis about the woman's problematic. It considers the woman social condition according to the androgynors ideology of the society. Briefly it presents a historical evolution of the woman in accord with the gender analysis. On the other hand, it presents too, an analysis of the health and the public politics in relation to the woman in Latin America, mainly in Peru, since the Incanato period, justifying the motives of the formulation and implementation of the Family Planning Program, the Responsible Paternity Program and Child and Mother Attention Program. PMID- 8716341 TI - [Reflections on the practice of breast feeding]. AB - The author makes analyses of breast feeding, doing a reflexion about the mainly indicators who has determined the actual situation, looking for identify ways for win the ideal cenary to future of breast feeding. PMID- 8716342 TI - [Views of nursing students on the curriculum development in night service]. AB - This preliminary work was based in the qualitative method, about the opinion of the College students nursing in the seventh (7th) stage of the course about the development of the curricular experience on the night shift. Thirty (30) relates were identified in the respective aspects: importance of the experiences, relationship between overseer nurse and multidisciplinary health care team; difference between day shift and night shift. The most part of the related demonstrates that it's been valid and they've given emphasis, only, in relationship between overseer nurse and multidisciplinary health care team, without emphasizing the effective contribution on the practice knowledge. The relating emphasized the difference among the shifts. It follows that is necessary an extense discussion of the teachers, nursing body and college students nursing body, under the effective objectives of the practice curricular experience for professional qualification. PMID- 8716343 TI - [The refusal of nursing consultation. A comprehensive analysis of the phenomenon]. AB - The purpose of this study was to understand the abandonment of nursing consultation focused on children 0 to 2 years old. A phenomenological approach was used to make apprehension of the essence of the phenomenon and to try to understand it without becoming attached to its determinant causes. Interviews were done with 12 mothers to describe their experiences on nursing consultation and, therefore, making possible to see the essence of the phenomenon. PMID- 8716344 TI - [Hospital infections in Brazil. Government actions for their control as expressions of social politics in the health area]. AB - The authors study the hospital infections evolution em Brasil and the demands for its control until its establishment by governmental actions. For a better understanding and analysis of the social actors' dispute process, their needs and interests and the relation with the governmental sector to the establishment of a certain state politics, a theoretical reference that enabled to elaborate the whole of social structure elements was searched. This theoretical reference should also take the filled of the relation between Society, State, Actors and Social Politics as its main question. In order to allude to these specific actions and its intervention range, it was necessary to make a distinction between the state politics modalities and the implementation responsibilities. Based on empirical material, it was possible to trace these demands, conforming on technological-assistencial models thorough the working process indicators. Other empirical categories levels was demanded to approach to the theoretical reference: specific conjectures, welfare medical assistance, the clinical intervention model and the quality movement. All the time, the analysis of the demands and the established project aimed the relation between the instruments and their purposes with themselves and with the health general politics, showing its implications. PMID- 8716345 TI - [Evaluation of the severity of head injuries using anatomical and physiological indices]. AB - The purpose of this study was the analysis of head injury severity based on physiologic and anatomic indexes, shown by hospitalized patients from the Intensive Care Unit. The study was characterized according to patients' age, gender, external cause and remaining time at the Intensive Care Unit, and it was verified that from the total of admitted patients the most of them were young adult males with a remaining time percentage of 6.28 days, Most frequent external cause was traffic motor vehicle accidents (65.72%). The study also considers the lesions and trauma severity according to the Abbreviated Injury Scale/ Injury Severity Score (AIS/ISS), reporting that the lesions were classified as severe lesions non-life threatening (AIS-3) and severe lesions life threatening (AIS-4). It was verified that the body region most affected was head and neck (57.65%). In relation to the trauma severity (ISS), most of the patients reached ISS 16. The study also presents the head injury severity by determining the consciousness level using the Glasgow Coma Scale (GCS); most of the patients showed severe head injury or GCS from 3 to 8. Analysis regarding a possible relation between the GCS and the AIS/ISS was performed to evaluate the head injury severity, considering the fact that the patient was victim of head injury or head injury and other body lesions. No meaning statistical element was observed regarding the relation between the GCS-1 and the AIS from the head region, except when analyzed by groups of severity, or GCS-1 3 to 8 and AIS-head 4 to 5; GCS-1 9 to 12 and AIS head 2 to 3. Similarly, no meaning statistical relation between the GCS-1 and the ISS, isolated head injury or head injury and other body lesions and the ISS or the GCS. It is expected that this study may contribute with new data for the continuous assistance improvement of patients, with head injury severity either related to other lesions or not, offered both by the Nursing and Health teams. PMID- 8716346 TI - [Situation of nursing students during their university courses appointed as crisis solvers]. AB - The study approaches the situations lived by the students of the various semesters of nursing in that University context, that, can often render negative feelings to the students, which are appointed as crisis and the actions developed in coping with difficulties. The method of collect was based answer on questions opened in order that the same ones could answer freely. The analysis of the testimonies carried in a classifying form thematic. The study showed the difficulties of the students concerning themselves, with the colleagues, of structural and financial order, etc. PMID- 8716347 TI - [An integrated approach to the planning of health promotion]. AB - This article addresses the specific problem of Health Promotion an Education planning, which involves three levels in an organization: the institution-wide level, the programmatic level and the user's level. Regardless of where the technical commitment is indicated, the ideal is to organize at all three levels, otherwise the program planning is likely to be incidental, fragmented, and ineffective. Comments are made on the theory of implementation and some barriers to implementation of Health Promotion and Education programs are discussed. PMID- 8716348 TI - [Comparative study of the indirect measurement of arterial blood pressure in pregnant women with a cuff of correct width and with a standard cuff]. AB - The hypertensive disorder of pregnancy is one of the most important complications of the pregnant cycle, presenting high risk of morbidity and mortality to the mother and the child. The marks of the systorical blood pressure and diastorical blood pressure are the parameters to diagnose the hypertensive disorder of pregnancy. Then, the precise measure of the blood pressure is an essential condition for the clinical evaluation of the pregnant woman during the antepartum course, in order to diagnose in advance, the hypertensive disorder of this period, since the raising in the blood pressure is normally the first clinical sign of the sickness (National High Blood Pressures Educational Program, 1990). The interest in the evolution of the blood pressure during the pregnancy is very old, since 1884, when Schedoff and Porockjakoff pointed out the attention to the association of the raising in the blood pressure with eclampsia (HENRY, 1936). Since then, many studies about the modification of the blood pressure caused by the pregnancy have been published, in order to recognize pathological diversions that characterize the hypertensive disorder of pregnancy. Although, the researchers are trying to identify an exact limit of the normal blood pressure during the pregnancy, there is a gap in the adaptation of the cuff to the circumference of the arm. In spite of the recommendation by the American Heart Association in 1951, which says that the width of the cuff should be 20% bigger than the diameter of the arm, no comparative study about the marks of the blood pressure during the pregnancy taken with a cuff of correct width and with a cuff of standard width, was found in the specific literature. (BORDLEY III et al., 1951) Comparing the indirect measure of the blood pressure taken with a cuff of correct width and taken with a cuff of standard width, ARCURI (1985) observed in 404 normotensive and not pregnant women between 17 and 59 years old underestimation of the blood pressure, especially in those women with thin arm. The co-relation is opposite to the circumference of the arm, what means that the smaller is the circumference the higher is the underestimation. Considering there is an underestimation of the levels of the blood pressure, when the large cuffs are used, it is called into question the jeopardy of the early diagnosis of high blood pressure of thin pregnant women. Based on the consideration presented, this study has the aim of evaluating the measurement of the blood pressure on pregnant women by using cuffs adequate to the circumference of the arm. The population of this study will be healthy pregnant women, with one alive fetus, registered in the antepartum course service of a maternity and of a community center in Sao Paulo city. PMID- 8716349 TI - [Breast feeding: assuming risks or guaranteeing benefits]. AB - The development of several action of promotion of the breastfeeding seems do not reach many of the variables that determinate the breastfeeding course. With the proposal of understanding what makes the woman decide about breastfeeding or wearing, this study was developed trying to get the conscious meaning of the breastfeeding experience for the woman, how the importance of the breastfeeding is shown by the woman's action and develop a theoretical sample that represents the experience of breastfeeding for the woman. Based on the theoretical presupposition of the Symbolic Interactionism and taking the Grounded Theory as the methodology, we described the breastfeeding based on the woman's point of view. The research shows that the woman experiences the breastfeeding from a process that evaluates and gives value to it, and that is understood by the themes: Evaluating the capability of breastfeeding, Evaluating the woman's feeling and Taking Decisions which the central phenomenon, representative of the process, is called WEIGHING THE RISKS AND BENEFITS. This study shows that the elements that give the policies for the breastfeeding result from the relation between the action of breastfeeding and several significant symbols shown in the interactive situations experienced by the woman. PMID- 8716350 TI - Predicting nurse staffing levels. PMID- 8716351 TI - Making our voice heard. Interview by Bill Doult. PMID- 8716352 TI - More demand but fewer nurses. PMID- 8716353 TI - It's good to talk. PMID- 8716354 TI - My hospital at home. PMID- 8716355 TI - Degrees of poverty. PMID- 8716356 TI - Who pays for his health care? PMID- 8716357 TI - Bearing the brunt of poverty. PMID- 8716358 TI - Ten steps to producing clinical guidelines. PMID- 8716359 TI - Caring for patients with motor neurone disease. PMID- 8716360 TI - Exploring confidentiality in nursing practice. AB - This article discusses the complex issues and dilemmas that nurses need to consider when dealing with confidential information. As well as reviewing situations where patients' rights to confidentiality conflict with public interest, the author discusses nurses' legal, ethical, moral and professional duties when handling confidentiality issues. PMID- 8716361 TI - Do birthplans empower women? A study of midwives' views. AB - Last week, we explored women's views of whether birthplans empower them in the birth process. This week, the second half of the study is presented. It explored the perceptions of empowerment from ten midwives, using the birthplan as a tool. Data were collected by semi-structured interviews. The findings indicated that several factors could disempower the women as well as the midwives. For women to be empowered, midwives need to provide open communication, adequate life skills, a nurturing and caring environment, and a democratic management structure. Advocacy and paternalism were found to be constraining factors for empowerment. PMID- 8716362 TI - Epilepsy (continuing education credit). PMID- 8716363 TI - Impressions of midwifery in New Zealand & Australia. PMID- 8716365 TI - Midwives' network. PMID- 8716364 TI - Thinking of working in New Zealand? PMID- 8716366 TI - A refreshing experience. PMID- 8716367 TI - Cambodia--daring traditional birth attendants to change tradition. PMID- 8716368 TI - Motherhood in Papua New Guinea. PMID- 8716369 TI - A tribute to hospice nurses. PMID- 8716370 TI - Hospice and skilled nursing facilities. PMID- 8716371 TI - An interrupted death. PMID- 8716372 TI - An interdisciplinary team approach toward death education. PMID- 8716373 TI - Enhancing volunteer effectiveness. AB - The characteristics of being genuine/congruent, having an unconditional positive regard and an empathic understanding, can be integrated into the training volunteers receive prior to assignment with patients/families. Understanding and practicing these concepts may further promote the interpersonal relationships volunteers have with patients/families and other hospice professionals. PMID- 8716374 TI - Oregon's assisted suicide law: a different perspective. PMID- 8716375 TI - Low-tech hospice home care for end-stage heart disease. PMID- 8716376 TI - SUPPORT study results--implications for hospice care. AB - This landmark study took place over a four year period of time and included 9,105 seriously ill hospitalized clients and their physicians. Its main objective was to improve end of life decision making and reduce the frequency of painful, mechanically supported, prolonged patterns of dying. Findings indicate that despite well planned, comprehensive interventions to improve study outcomes, clients in the intervention group fared no better than their counterparts in the control group. These findings have serious implications for those working in hospice and palliative care. PMID- 8716377 TI - The effect of an alphanumeric paging system on patient compliance with medicinal regimens. AB - There are numerous actual and potential impediments to patient compliance with medicinal regimens. The implications of poor compliance include worsening disease states or symptoms, with resultant increased costs of care. Using a population of hospice patients at high risk for noncompliance, this study evaluated the use of an alphanumeric paging system as a memory prosthetic, finding that compliance rose from a mean of 56 percent to 96 percent when the system was used. We conclude that such a program is of use in this setting, and that these findings warrant study and suggest applications in other populations. PMID- 8716378 TI - Ethical aspects in palliative care. PMID- 8716379 TI - Hepatitis A & B: the nurse's role (continuing education credit). PMID- 8716380 TI - Local and systemic effects of major traumatic wounds. AB - Major traumatic wounds cause a chain of complex localized and systemic events. Understanding these physiologic processes, potential complications, and treatment priorities can assist the nurse in facilitating wound healing and optimizing patient outcomes. PMID- 8716381 TI - Wound healing and critical illness. AB - Critically ill patients with acute wounds provide a multitude of challenges for the critical care nurse. Assessment of the patient's wound and associated-disease risk factors must be an ongoing process involving the evaluation of the treatment modalities and the stage of wound healing. Nightingale stated that the nurse's role "is to put the patient in the best condition for nature to act upon." The critical care nurse plays a pivotal role in establishing and maintaining the best condition or environment that will maximize the body's process of wound healing and expedite the patient's return to a state of wellness. PMID- 8716382 TI - Wound assessment and decision-making options. AB - The increasing presence of multiple, chronic disease states among critically ill patients compounds care of their acute or traumatic conditions. These patients may be admitted with existing wounds and they may develop impaired skin integrity due to a variety of factors related to their illness and treatment. Whereas maintenance of vital functions is the priority of care, critical care nurses are more frequently faced with the challenges presented by pressure ulcers and various other surgical, traumatic, and vascular wounds. These wounds increase morbidity and mortality and result in higher costs for health care. Education in assessment of risk, pathophysiology, the healing process, comprehensive assessment, and efficient, effective wound management contribute to optimal patient outcomes and cost-effective delivery of care. PMID- 8716383 TI - Using wound care products to promote a healing environment. AB - Healing is characterized by the synthesis of new tissue and scar formation. Despite the complexity of healing with full-thickness injury, the repair process occurs in a predictable manner. There are four basic principles of wound care: (1) debride necrotic tissue and cleanse the wound to remove debris, (2) provide a moist wound healing environment through the use of proper dressings, (3) protect the wound from further injury, and (4) provide nutritional substrates essential to the healing process. Most importantly, any underlying pathophysiology must be treated or the wound will not heal. Products selected to create a healing environment must be chosen thoughtfully and scientific rationale must support their use. Intensive care nurses have the opportunity to get the patient off to the right start by attending to the basic principles elucidated in this article. Accurate wound assessment and appropriate product choices can promote a healing environment. Intensive care of patients includes differentiating wound types and making appropriate wound care product decisions that ultimately affect patient outcomes. PMID- 8716384 TI - Nutritional needs of the acutely ill with acute wounds. AB - The purpose of nutritional support in the critically ill is to blunt the effects of the stress response, to preserve musculature and organ function, and to promote hepatic protein synthesis, tissue regeneration, and wound healing. Adequate nutritional support is vital to the patient with acute wounds. This article addresses the body's response to physiologic stress and reviews means of supporting nutritional status and the substrates used to maximize nutritional therapy. The critical care nurse will have a better appreciation for how and when to implement nutritional therapy, appropriate nutrients, and methods for monitoring nutritional status. PMID- 8716385 TI - Surgical options in wound healing. AB - This article addresses methods of reconstruction for major acquired and traumatic soft-tissue defects in adults. The reconstructive ladder is used as a basis for discussing the various options for surgical closure. Conditions such as facial trauma, head and neck cancer, median sternotomy, sternal osteomyelitis, bronchopleural fistula, necrotizing fascitis, pressure ulcers, and degloving injuries are used as examples of reconstruction. Nursing aspects, including postoperative flap monitoring, are reviewed. Outcomes of reconstruction, including flap failure, are addressed. PMID- 8716386 TI - Psychological considerations for patients with acute wounds. AB - The range of psychological considerations for patients with acute wounds is very great, to an extent that makes any generalization extremely difficult. Adverse psychological developments seem to occur most often when there are permanent changes in the body's structure or function. Reactions may be particularly troublesome if the wound occurs as a result of trauma, involves an emotionally loaded body part or function, or results in visible deformity. Critical care nurses can help patients begin the process of reintegrating the body image by providing congruent verbal and behavioral messages that communicate to the patient that his or her worth and value as a human being has not been lost. PMID- 8716387 TI - Diabetes and wound healing. AB - The effect of diabetes on wound healing is a very complex issue that continues to leave many unanswered questions. There continues to be an empiric fact that diabetic patients are at increased risk for the development of infection and impaired wound healing. The precise underlying defect(s) continue to be a focus of research; however, it is generally agreed that improved glycemic control plays a significant role in lowering the risk of wound healing complications. Diabetic patients need to pay special attention to their diabetes care in general as well as to the critical illness, wound, or ulcer that has brought them to our care. PMID- 8716388 TI - Advances in burn wound care. AB - The development of effective topical chemotherapy, the reintroduction of burn wound excision, and the use of biologic dressings have significantly decreased the incidence of invasive burn wound infection and have contributed to the improvement in the survival that has occurred over the past four decades. The currently available skin substitutes are imperfect, and research endeavors continue in the effort to develop a nonantigenic disease-free, readily available, physiologically effective tissue that will promptly effect wound closure, reduce scar formation and thereby improve cosmetic results, and reduce the need for reconstructive surgery. As monitoring and physiologic support techniques improve and additional advances in wound care occur, the morbidity and mortality of burn patients will be further reduced. PMID- 8716389 TI - Outpatient wound care. AB - As patients are discharged from the acute care setting to the home care setting at a much earlier time in their recovery, surgical wound care will be delivered by patients, family members, and home health care providers rather than by the hospital nurse in a traditional in-patient setting. This trend, which began in the mid-1980s, is expanding to include not just surgical wounds but also surgical complications such as wound dehiscence and traumatic wounds healing by secondary intention. Intensive care nurses are involved in discharge planning from the time the patient is admitted to the ICU. Early planning and teaching regarding wound care, universal precautions, and medical waste disposal have become a vital component of preparing the patient for optimal continuity of care as the transition is made into the community. PMID- 8716390 TI - Guidelines on resuscitation pass judicial test. PMID- 8716391 TI - Where have all the students gone? PMID- 8716392 TI - Doctors' assistant or a trojan horse? PMID- 8716393 TI - Need to know. PMID- 8716394 TI - Conned for the last time. PMID- 8716395 TI - Reliving the nightmare. PMID- 8716396 TI - Taking the lead. PMID- 8716397 TI - Stolen by aliens. PMID- 8716398 TI - Promoting research and education in dementia care. PMID- 8716399 TI - Telephoning a nursing department: callers' experiences. AB - This preliminary, exploratory study examined the level of satisfaction of patients and relatives with the telephone communication skills of nurses. Results indicate that callers experienced several difficulties, particularly with regards to knowing who they were speaking to, being treated as an individual rather than just another caller, having their calls dealt with efficiently, or redirected correctly. It is suggested that nurse educators include training in telephone use in courses on communication skills. PMID- 8716400 TI - Caring for surgical patients with sickle cell disease. AB - This article aims to enlighten nurses working on surgical wards and in the operating theatre to give appropriate and quality nursing care to patients with sickle cell disease. The author considers patients who experience a vaso occlusive crisis. PMID- 8716401 TI - Developing a direct access outpatient hysteroscopy service. AB - This article describes the development and running of an outpatient hysteroscopy service. The authors highlight the reasons why the service was seen as feasible and the benefits which patients and staff have found since its inception. PMID- 8716402 TI - Linking knowledge and practice through teacher-led placements for students. AB - Project 2000 and diploma level education for pre-registration students have been criticised for their lack of emphasis on practical nursing skills. This article describes how teacher-led placements can equip these students with the requisite practical skills in a reflective manner that enables them to relate theory to practice. The author argues that only through critical thinking can students obtain a relevant understanding of the knowledge which underpins both theory and practice. PMID- 8716404 TI - The right image: an important ingredient in success. PMID- 8716403 TI - Dysphagia in motor neurone disease (continuing education credit). PMID- 8716405 TI - The reminiscing game. PMID- 8716406 TI - A story of recovery and second chances. PMID- 8716407 TI - Putting the conceptual model into the research report. AB - Writing research can be challenging--especially writing the conceptual model. Deciding how much to include of the conceptual model and where to place it is difficult. This experienced research author presents four steps to guide the nurse author in writing the conceptual model for research reports. PMID- 8716408 TI - In-house nursing publication: tips on troubleshooting. AB - Many nurse authors and editors are involved in the production of newsletter publications for their hospital, health-care organization, or association. It is a labor-intensive job that can be very rewarding; however, all publications have problems that arise--no articles, poorly written ones, need for extensive editing, copyright concerns, and the administrative details of typing, typesetting, selecting paper and ink, missed deadlines, typos, or publication of the wrong names or titles. PMID- 8716409 TI - Comparison of the effectiveness of tutored videotape instruction versus traditional lecture for a basic hemodynamic monitoring course. AB - As nurse educators find themselves faced with increasing responsibilities and as nursing administrators seek to use nurse educators in the most cost-effective, resource-efficient manner, alternatives to the traditional lecture must be sought. Tutored videotape instruction is an educational method in which a tutor serves to clarify points of confusion and to stimulate discussion during a videotape presentation. Using a hierarchal nested research design, there were no significant differences in participant preference or posttest knowledge scores between the methods. As a result, tutored videotape instruction was adopted as an acceptable educational method. In addition, tutored videotape instruction can serve as a developmental tool for RN preceptors as tutors and for newly hired critical care educators. PMID- 8716410 TI - Working with lesbian, gay, and bisexual people. Reducing negative stereotypes via inservice education. AB - Basic components of an inservice program that concerns working with lesbian, gay, and bisexual people are presented. Staff nurses will encounter such people in every nursing specialty and will have lesbian, gay, and bisexual coworkers whether they know it or not. Inservice programs that deal with any sexuality issue must include cognitive and affective interventions, because negative or uncomfortable attitudes may have been ingrained since childhood. PMID- 8716411 TI - A comprehensive plan to meet the unit-based education needs of nurses from several specialty units. AB - Nursing staff development departments across the country have responded to the anticipated changes in health care in a variety of ways. This need to reexamine the traditional role of the clinical instructor has resulted in the development of creative strategies designed to meet the unit-based education needs of nurses. In this article, the authors describe a comprehensive approach that not only meets the learning needs of staff members from several specialty units but also fosters their professional development. PMID- 8716412 TI - Examining competency-based orientation implementation. AB - A competency-based orientation program provides a structured, comprehensive approach to the process of orientation. However, the program is only as good as the implementation. It is important for management, preceptors, and orientees to be knowledgeable of the details of the program and the procedures involved in the implementation process. when problems arise, it is important that all those involved solve the problems in an organized, systematic fashion. PMID- 8716413 TI - Evaluation of a hospital staff's knowledge regarding the use of physical restraint. AB - A hospital nursing staff's understanding of policy and practice when using physical restraints and safety devices was evaluated using the knowledge test contained in a restraint research project designed at the University of New York at Buffalo. Although overall understanding of restraint use and policy was good, this evaluation pointed out areas of need that will be addressed in future education programs. PMID- 8716414 TI - Quality of decision making by registered nurses. AB - This study investigated quality of decision making in a convenience sample of 26 registered nurses. Instruments used were the Joseph Decision Making Tool, Actual Decision Making instrument, and computer-assisted clinical simulations. Results were correlated with educational preparation, age, experience, practice area, and specialty certification. No differences among basic educational preparation with frequency of actual decision-making practices and willingness to make decisions were found. A positive correlation was found between practice area and frequency of actual decision making (r = 0.639, P < 0.001), and practice area and communication (r = 0.439, P < 0.05). PMID- 8716415 TI - Developing management skills: an experiential exercise. PMID- 8716416 TI - Planning a continuing education conference. PMID- 8716417 TI - Dilemmas. PMID- 8716419 TI - Sadness, tragedy and mass disaster in Oklahoma City: providing critical incident stress debriefings to a community in crisis. AB - Shortly after 09:00 h on 19 April 1995, the Alfred P. Murrah Federal Building, located in downtown Oklahoma City, was devastated with a bomb blast of such gigantic proportions that it was heard 60 miles away in neighbouring Norman, Oklahoma. Oklahomans routinely commuting to work on that sunny Wednesday morning went about their business as usual. A crude bomb chemically comprised of various organic compounds, chemical fertilizer, ammonium nitrate and diesel fuel, weighing an estimated 4800 pounds or more, was transported in a vehicle the size of a truck. It blew open a crater 6-8 ft deep in the street floor. The Murrah Federal Building was impacted immediately; floors, windows, communication equipment and almost all the innocent victims inside were razed to the ground. Outside the building, as far as 10 blocks away or more, hundreds of victims lay hurt, seriously injured or dead from shards of glass that flew from office windows hundreds of feet above the street floor. Without warning, the initial impact of the bomb immediately devastated the entire city. People were in a state of shock, disbelief and denial; acute symptoms of post traumatic stress disorder (PTSD) were commonplace. Oklahomans, 'numb' from the impact of the critical incident and ill-equipped to handle the chaos of such catastrophic proportions, struggled to regain control of their lives as friends, family and loved ones went unaccounted for or were found critically injured, dying or already dead. The critical incident on 19 April demanded the immediate attention of the nation, to come to the aid of the Oklahomans who were in desperate need. By 1 June, the exhaustive investigations revealed that 30 office buildings in downtown Oklahoma City had to be condemned, and as many as 300 others were damaged. In addition, 168 people had been found dead including 19 children and one nurse working as an emergency services rescue worker. Approximately 490 other victims had been reported injured from the blast. Countless others have been traumatised by the critical event and will need professional attention and care for weeks, months and years to come. The final extent of the impact on Oklahoma City is yet unknown and may never realistically be estimated in terms of personal trauma, loss and grief. PMID- 8716420 TI - Does debriefing after psychological trauma work? Time for randomised controlled trials. PMID- 8716422 TI - The Muslim Law (Shariah) Council and organ transplants. PMID- 8716421 TI - Knowledge of paediatric first aid in the general population. AB - If accidents cannot be prevented or ameliorated by protective devices, good first aid is essential to reduce the further effects of an injury. This paper highlights deficiencies in first aid knowledge among a random selection of the general population in Edinburgh. The results of this study indicate that first aid knowledge is generally deficient, with only 13% of people able to correctly treat all paediatric emergencies described. Recognised first aid courses, such as those run by voluntary agencies, focus their training on adult injuries, adapting care for children from this. There is no specific paediatric first aid course run by a recognised authority. Recommendations to improve this are made. PMID- 8716418 TI - Ophthalmic trauma in accident and emergency. AB - Many nurses and doctors have little ophthalmic experience and this leads, inevitably, to a lack of confidence when presented with patients with ophthalmic problems. Six per cent of all trauma presenting to the Accident & Emergency (A & E) department is ophthalmic and this, combined with a knowledge gap, can lead to potential management problems for a high number of patients. Most presenting ophthalmic trauma is minor in ophthalmic terms, if not to the patient, but the early identification of more serious problems is essential in order that vision is not permanently damaged. This article addresses some of the more common, minor injures which are seen in the A & E department and considers their presentation and treatment. The assessment and recording of visual acuity is addressed in detail as the main indicator of visual function and an essential component of any eye examination. The use of eyepads in the treatment of eye injury is also considered, as are issues surrounding the use of eyedrops. The article concludes with the suggestion that ophthalmic units be approached for advice about ophthalmic problems in order to further enhance the knowledge of A & E staff. PMID- 8716423 TI - Plaster checks by nurses: safe and efficient? AB - This prospective, blinded comparison study compared plaster checks performed by nurses with those performed by doctors. Nurses performed this task well. Allowing plaster checks to be performed by nurses, with referral to a doctor if they are concerned, would have reduced the number of plaster checks performed by doctors by two-thirds. Institution of this process could result in reductions in waiting time for patients presenting for plaster checks and in improved efficiency in the Emergency department. PMID- 8716424 TI - Education in emergency nursing: a new beginning. Part I. AB - The movement of nurse education to the tertiary sector is all but complete in Australia. Trends are changing as new partnerships are being forged and the focus of education is now responsive to economic and political factors. These factors have resulted in the regionalisation of health care facilities and the de institutionalisation of health services in Queensland, Australia. Moreover, de institutionalisation of services coupled with economic rationalism has pre-empted the offering of post-graduate clinical courses by university nursing facilities. This paper addresses the process of developing the content of new courses under the umbrella of the Graduate Diploma/Masters in Emergency Care Nursing offered at Griffith University Gold Coast, Australia. This initiative is the first in Queensland and has revisioned the education of emergency care nursing. This course includes a tapestry of knowledge enhancing, clinical and factual subjects presented by experienced clinicians teaching with academics, recognising the collaborative nature of nursing practice with academics and clinicians working jointly in course development, teaching and clinical placement. Strategies developed collaboratively addressed course content including logistics, time management, the critical selective selection of appropriate content, assessment criteria, registerability within Queensland and other states and clinical competence. Moreover, this process of developing the course utilised the Australian National Registering Authorities' Competencies and Benner's use of the Dreyfus model to guide the knowledge level of the clinician at course completion. In particular this paper will not only address the blending of theory and practice to consolidate the relationship inherent between the clinician and education, but will establish the fact that no gap exists. PMID- 8716425 TI - The lifeline of triage. AB - Most acute hospitals operate a triage system to evaluate and prioritize the care and needs of clients presenting in the Emergency department. The emergency nurse, therefore, employs a variety of cognitive processes to clinically categorize clients in relation to the most acute need. This ability to clinically assess is a combination of knowledge and intuition grounded in experience and formal education. Therefore, the optimal functioning of the Emergency department is dependent upon not only the protocols and policies of hospitals, but the skills, experience and the confidence of individual triage nurses. This paper will reconsider the role of the triage nurse by addressing a literature deficit related to triage and the triage nurse, as the front line mediator, who is in the prime position of being able to rise the professional profile of the emergency nurse by making visible the care provided by nurses in this department and, most significantly, in the triage waiting room. This care extends beyond pointing to the waiting room clock and a sign which informs clients of the purposes of the emergency care department. This paper will also discuss a range of strategies used by the authors to enhance effective triaging to ensure a quality standard in client care. PMID- 8716427 TI - Did I do the right thing? An accident and emergency nurse. PMID- 8716426 TI - Sudden bereavement in accident and emergency: the development and evaluation of a short educational course for qualified nurses. AB - Qualified nurses working in the Accident and Emergency (A & E) department are in a unique position to assist family members following the crisis event of sudden bereavement. However, there is evidence that many nurses lack specific educational preparation for this demanding role. This paper describes the development of a short course designed to meet the needs of A & E nurses with a variety of experience of working with the bereaved. Particular reference is made to content and the use of experiential teaching methods, as opposed to more didactic approaches. The results of a self administered questionnaire (n = 30) designed to evaluate the first three courses run, which was returned by the participants 4 weeks after the end of each course, are presented. A majority of the sample reported increased knowledge and confidence regarding the management of sudden bereavement following the courses. Twenty-seven (90%) of the respondents stated that their personal anxiety about death and dying had decreased. Some concern was expressed, however, about the particular value or role play as a teaching method. A number of possible explanations for this are discussed in the light of the literature. PMID- 8716428 TI - The organ donation process through a review of the literature. Part 2. AB - Several studies have attempted to investigate the role of culture and race in organ donation and transplantation. These studies go beyond ones conducted out of mere interest. The researchers have found that there are indeed human leukocyte antigens that are specific to individual races and that affect transplantation outcomes (Milford et al 1987). Similarly, Weller et al (1987) have shown that graft and patient survivals were improved when donors and recipients are of the same race. PMID- 8716429 TI - [Ethical aspects in nursing research]. PMID- 8716430 TI - [A contribution for the bridging of the chasm between research and practice. Why is nursing research so important?]. PMID- 8716431 TI - [When poverty becomes a disease. A big step into a better future]. PMID- 8716432 TI - [Treatment care and nursing insurance]. PMID- 8716434 TI - [Enteral feeding. II. Advantages and problems for those involved]. PMID- 8716435 TI - The betrayal. PMID- 8716433 TI - [Comparison of networking models]. PMID- 8716439 TI - Roles revamp. PMID- 8716438 TI - Heart to heart. PMID- 8716436 TI - Paying the price for long-term care. PMID- 8716437 TI - Healthy debate. PMID- 8716440 TI - Starting from scratch. PMID- 8716441 TI - Research and development in forensic mental health. PMID- 8716443 TI - Nursing care of the patient with a tracheostomy. AB - Patients who require a tracheostomy have particular needs associated with their care to ensure their safety and well-being. In this article, the author describes the reasons why a patient might have a tracheostomy and provides a detailed account of the anatomy, care and complications associated with it. PMID- 8716442 TI - Are nurses independent and autonomous practitioners? AB - The purpose of this paper is to review the present position of nursing as a developing profession. The author considers the forces stimulating change and the extent to which nurse practitioners hold professional independence and practise nursing from an autonomous base. PMID- 8716444 TI - Patient focused menu planning: an ENB project in a hospice. AB - This paper discusses how a small study of the meals provided for patients in a hospice was undertaken as part of the ENB 931 Care of the Dying course, and used to change practice. The authors describe how the study provided not only useful information about the types and aspects of food both enjoyed and disliked in this setting, but also stimulated interest among nursing and kitchen staff, resulting in a programme of changes in menu planning. It is argued that while the project was undertaken as a piece of research, it actually had more in common with audit, and that audit might be a more useful activity to incorporate into ENB courses. PMID- 8716445 TI - Resuscitation: basic life support (continuing education credit). PMID- 8716446 TI - Power nap rejuvenates body, mind. PMID- 8716448 TI - Nursing education administrator unravels confusion about CNE. PMID- 8716466 TI - Acronym acrimony. PMID- 8716453 TI - Back to life. Interview by Andrew Cole. PMID- 8716469 TI - The perceived impact of committee participation on job satisfaction and retention of staff nurses. AB - In 1989, in response to the nursing shortage and much literature on dissatisfiers for practising nurses, the Ontario Minister of Health introduced a new regulation to the Public Hospitals Act, requiring that staff nurses and nurse managers have widespread representation on hospital committees. While involvement of nurse managers on such committees was not new, the participation of staff nurses certainly was. PMID- 8716458 TI - Making sense of radiotherapy: curative and palliative. AB - Radiotherapy is one of a range of treatments available for the treatment of cancer. It involves the use of ionising radiation to interrupt cellular growth in malignancy. Its use can be either curative or palliative. This paper discusses how radiotherapy works, how it is applied and introduces its side-effects. A subsequent paper will look in more detail at side-effects and nursing care. PMID- 8716470 TI - Analysis of powerlessness in nursing work. AB - Despite the plethora of articles on work environment and nursing power in the recent nursing literature, we have yet to improve nurses' experience of work in any significant way. In fact, as hospital budgets come increasingly under attack in the name of health care reform, nursing work is becoming even more demanding and, to many, less satisfying This article describes the experience of powerlessness by a sample of hospital staff nurses in British Columbia. It analyses the themes which characterize powerless work situations and explores the impact of these on nurses. It suggests some ways of bringing about change in these circumstances of work. The article intends to illuminate, for those who care to understand, the complexity and often insurmountable challenges that nurses encounter in the course of their nursing work. It urges us to create opportunities to make the increasingly difficult and frustrating circumstances of nursing work visible to key decision makers. PMID- 8716460 TI - Working for the good life in rural west Somerset. AB - This paper outlines a community development project in rural Somerset that was instituted in response to identified health needs in the area. Organisational changes meant that funding for the project was curtailed so that completion of the planned programme and full evaluation could not be carried out. PMID- 8716457 TI - Crossing the line. PMID- 8716471 TI - Changing nursing practice--trisectoral collaboration in decision making. AB - In an age of cost containment, agency partnerships have become an essential element for future planning and program implementation. This paper describes a trisectoral collaboration of a hospital, health department, university and school of nursing to compare the efficacy and efficiency of referral decisions of hospital staff nurses to those of the public health liaison nurses (LNs). A process to identify decision criteria was undertaken and an educational programme was designed to assist the staff nurses with the referral process and to assure consistency of decision making. The two groups were then compared. The results of the study found staff nurses, using the decision criteria, identified more patients who required public health nursing visits than did the liaison nurses, refusal rate of the patients to participate was no different, staff nurses cost less than LNs and job satisfaction was not significantly altered for either group. In addition to providing information to guide administrative and clinical decision making, the project also provided a learning experience for the staff of three agencies in conducting research and in using evidence-based practice to change traditional practice. PMID- 8716461 TI - Guidelines and protocols: a chance to take the lead. AB - Protocols and guidelines have been heralded by many as a panacea for the problems associated with the rising costs of health care and variations in clinical practice. These variations occur despite very similar medical conditions and often indicate poor use of research-based evidence to inform patient care. This has resulted in a national push towards greater use of protocols and guidelines as means of informing and indeed improving clinical practice. In order that protocols and guidelines achieve their full potential, it is essential that nurses, who deliver the majority of direct patient care, grasp the opportunity to influence their development. They need to be used as a framework to guide existing practices and as a focus to shape future work on clinical effectiveness. This paper examines definitions of guidelines and protocols and considers their use for the development of nursing practice, improvement of patient care and introduction of greater clarity in the use of health-care resources. PMID- 8716447 TI - ANA calls for safety, quality measures. PMID- 8716451 TI - Anxiety and depression in older adults: an independent study (continuing education credit). PMID- 8716464 TI - Complementary therapies. Sleepless and unsettled. PMID- 8716465 TI - Pressure relief: a guide to what is on offer. PMID- 8716462 TI - New breast-feeding standards used as tool of change. AB - This paper describes a breast-feeding standard that was set and audited in the early stages of a trust's midwifery project five years ago. It describes the audit cycle as an example of how care can be improved through standard setting and as a way of highlighting issues for consideration. PMID- 8716454 TI - Life can go on after a serious back injury. Interview by Andrew Cole. PMID- 8716450 TI - When to use a letter only--without a resume. PMID- 8716449 TI - IOM study on nursing staffing falls short. PMID- 8716456 TI - Failing to turn the world upside down. PMID- 8716467 TI - Learning disabilities. Confusing diagnoses. PMID- 8716474 TI - RCN's conflict over its professional and trade union responsibilities. PMID- 8716473 TI - Registered nurses' perception of their autonomy and the factors that influence their autonomy in rehabilitation and long-term care settings. AB - To determine attributes of the registered nurse and the working environment that influence registered nurses' perception of autonomy (defined as independence within one's own practice), data were collected from questionnaires completed by 85 full time registered nurses at a 601 bed chronic/rehabilitation hospital located on two sites in the city of Toronto, Ontario, Canada. Registered nurses experienced a higher perception of autonomy given a more participative management style on the part of their nurse managers. Place of basic nursing education, which was used as an indication of cultural origin, was found to contribute to registered nurse perception of autonomy where basic education was in Canada or the United States. Increased workload (acuity) was found to detract from registered nurse perception of autonomy. Baccalaureate education, and primary or modular nursing care delivery models contributed to high autonomy, whereas diploma education, and functional or team nursing were associated with the low autonomy group in discriminant analysis. Administrative recommendations for nursing include the promotion of a participative management style on the part of nurse managers; attention to cultural diversity of nursing staff; support of advanced education in nursing; support of primary or modular nursing care delivery models; and management of registered nurses' workload in order to encourage a balance between client-focused professional practice activities and the routinized, provider-focused, task-driven activities. PMID- 8716476 TI - Metastasis: the deadly part of cancer. AB - The most malignant characteristic of any cancer cell is the cell's ability to metastasize. The movement of the malignant cell from the primary neoplasm to a distant site, and the cell's subsequent growth there, is the most dangerous characteristic of any tumour cell. Metastatic development occurs via a series of complicated events. This article explores the stepwise process of metastatic spread, known as the 'metastatic cascade', highlights the organs commonly involved in metastatic spread, and debates the seed and soil hypothesis of metastasis. The nurse is introduced to the importance of recent medical research which highlights the genetic links identified in metastatic cell formation. Tumour suppressor genes and oncogenes, which are genes known to induce or promote cancer, and their implications for prognosis, are explained. Finally, the nurse's responsibility in maintaining his/her knowledge and understanding of metastasis in order that he/she may provide appropriate care and support for the patient and family is discussed. PMID- 8716459 TI - Functional bracing of tibial shaft fractures. AB - This paper examines the reduced risk of non-union in tibial fractures by allowing controlled motion at the fracture site. It argues that this can also stimulate healing and can be achieved by using functional bracing, which holds the position of the limb, allows weight-bearing and mobilisation of neighbouring joints. It also describes how to apply such a brace. PMID- 8716463 TI - The modest giant of medicine. PMID- 8716477 TI - Intraperitoneal chemotherapy: a prolonged infusion of 5-fluorouracil using a novel carrier solution. AB - A novel carrier solution, icodextrin 20 (7.5%) has allowed exploration of prolonged intraperitoneal (IP) infusion of the cytotoxic drug, 5-fluorouracil. Eighteen patients with intraperitoneal carcinomatosis were entered into a feasibility and pharmacokinetic study of prolonged regional (IP) chemotherapy.. Specialist nurses trained the patients to self-administer their own treatment via a permanent i.p. catheter. A twin bag delivery system was used to perform one exchange daily. It proved possible to deliver continuous (5 days per week) i.p. 5 fluorouracil at doses of 200 mg/m2 and 300 mg/m2 for up to 12 weeks. The toxicities seen were infective peritonitis, nausea and vomiting, lethargy and anorexia. This was a nurse-led study and the home-based therapy holds promise for patients with malignant peritoneal disease. PMID- 8716478 TI - Causation of problematic behaviour in people with learning disabilities: 1. AB - This article, the first of two parts, examines the need for learning disability nurses to explicate the specific behaviours which are often grouped under the umbrella term 'challenging/problematic behaviours'. The prevalence of such behaviours in people who have learning disabilities is discussed. In addition, a review of the main theories of causation of problematic behaviour is presented, with a particular emphasis on the issues of violence and aggression in clients. The second article will offer suggestions on interpreting challenging behaviours in people who have learning disabilities. PMID- 8716472 TI - Staff nurses' perceptions of job empowerment and level of burnout: a test of Kanter's theory of structural power in organizations. AB - Kanter's structural theory of organizational behavior was used as framework to explore the relationship between perceptions of power and opportunity and level of burnout in a sample of 87 hospital staff nurses. Data were collected using a modified version of the Conditions for Work Effectiveness Questionnaire (Chandler, 1986) and the Human Services Survey (Maslach & Jackson, 1986). Consistent with Kanter's theory, perceived access to power and opportunity was significantly related to the three aspects of burnout: level of emotional exhaustion and depersonalization (r = -.3419, p = .004; r = -.2931, p = .02), and personal accomplishments (r = .3630, p = .002). The results of this study are useful for nurse administrators positioned to create organizational structures than empower staff nurses and subsequently decrease burnout. PMID- 8716479 TI - Accepting the challenges of pain management. AB - Despite advances in the treatment of pain, studies reveal that hospitalized patients continue to suffer under-recognised and unrelieved pain. Several factors contribute to this problem including health-care professionals' lack of knowledge about pain and analgesia, the inadequate use of pain assessment techniques and incomplete charting of patients' reports of pain. This article describes these challenges which must be overcome if the situation is to improve. It outlines the potential value of systematic assessment and documentation of pain so that the patient's pain is acknowledged as a unique experience. This should enable the health professional to individualize his/her approach to pain management. PMID- 8716455 TI - When training is a world apart. PMID- 8716452 TI - Specialist nurses face axe in AIDS care cuts. PMID- 8716481 TI - The basic experimental design. AB - This article considers the three characteristics of the basic experiment (random assignment of subjects, manipulation of relevant variables and control of irrelevant variables) and describes the two most common experimental designs: the pre-test/post-test design and the post/test only design. The necessity of controlling for subject and situational variables is highlighted and the challenge researchers face when using experimental approaches in other than laboratory situations are addressed briefly. PMID- 8716468 TI - Learning disabilities. Your case or mine? PMID- 8716475 TI - Infection control: human T-cell leukaemia virus infection. AB - Human T-cell leukaemia (HTLV) types I and II are blood-borne viruses which are transmissible by sexual contact, transfusion of infected blood, sharing equipment for intravenous drug use and breast-feeding. At present the estimated seroprevalence of these viruses in the UK is approximately 1 in 20,000. HTLV I/II infection carries a small and unpredictable risk of leukaemia or paralysis. In the absence of a treatment for HTLV infection, or effective therapy for associated diseases, the prevention of further transmission is of primary importance. Patients need sensitive and appropriate advice in order to understand the implications of their infection. PMID- 8716482 TI - Classification of pressure sores: 2. AB - Many different pressure sore classification scales are used in the UK. They all have differing strengths and weaknesses which may affect their use in research studies, prevalence and incidence surveys, and collaborative care planning. This article the second of a two-part series, examines which scales fit the NHS Executive's (1995) recommendations of practice guidelines for the classification of pressure sores and discusses what evidence is available relating to the validity, reliability and utility of existing classification scales. None of the scales comply fully with the practice guidelines' recommendation that they should be applicable to all skin types, and only two scales have demonstrated fair levels of inter-observer reliability. PMID- 8716480 TI - Elderly people's knowledge of and attitudes to pain management. AB - Pain is a frequent complaint of elderly people in hospitals and in the community, yet it is often not managed effectively. Barriers to effective management have included fear of narcotic addiction, inadequate assessment of pain, and attitudes of health professionals. Attempts have been made to improve the knowledge and attitudes of health-care professionals. This study examined the attitudes and knowledge of acute pain and pain management among well elderly people in rural and urban settings in the southeastern USA in order to identify possible barriers to effective pain management. Questionnaires on knowledge and attitudes towards pain and pain management were given to 62 rural and 63 urban well elderly people aged 65 years and older. Findings demonstrate a lack of knowledge as well as the presence of non-facilitative attitudes about pain and pain management. PMID- 8716483 TI - The Modular Propad and the Softform mattress. AB - Constant low-pressure supports such as soft mattresses can feel comfortable because they conform well to the shape of the body. When such mattresses are provided with a highly convoluted surface made up of contiguous foam pillars, their efficacy is improved. The Modular Propad is a highly convoluted, but shallow, foam mattress. It is used as an overlay for an ordinary mattress and seems to be superior to earlier forms of convoluted foam overlay. The Softform is a new mattress which has replaceable sections. The outermost sections of the Softform are convoluted foam sheets similar to the Modular Propad. This article discusses the advantages and disadvantages of these products and some potential problems with their covers. It also suggests that the use of a pressure sore risk assessment tool can facilitate the allocation of patients to these special supports. PMID- 8716485 TI - Ask yourself, "Why do I stay in oncology nursing?" and think about your response. PMID- 8716484 TI - Extremes of the nurse practitioner role. PMID- 8716487 TI - Fighting back with a badge of honour. PMID- 8716488 TI - Has anyone seen their top-up pay? PMID- 8716486 TI - Time to heal. PMID- 8716489 TI - Equal opportunities. Everybody's problem. PMID- 8716490 TI - Equal opportunities. Race into action. PMID- 8716491 TI - Equal opportunities. Warning: racism at work. Interview by Janet Snell. PMID- 8716492 TI - The impact of primary nursing on patient satisfaction. AB - Primary nursing is considered by some to be an appropriate vehicle for meeting the named-nurse standard of the Patient's Charter. The study reported here set out to determine whether patients identifying one nurse as being in charge of their care would express more positive experiences of and greater satisfaction with nursing care. Patients who could identify a nurse in charge of their care reported more positive experiences of that care. However, in the wards studied, primary nursing did not appear to be any more successful than team nursing in encouraging this process. Given the limited resources to implement primary nursing, the authors advise increased efforts to ensure each patient has one nurse responsible for their care, and that they know who this nurse is. PMID- 8716493 TI - Should people with HIV infection receive transplants? AB - This paper examines whether potential organ recipients should be screened for HIV/AIDS. Written by a former staff nurse at a liver unit, it outlines the results of a survey of transplant centres. The author argues for pre- and post test counselling and calls for a protocol to be set up as a guide to testing. PMID- 8716497 TI - Buyer beware. PMID- 8716495 TI - An evaluation of the role of the nurse practitioner. AB - There is a lack of clarity over where the role of the doctor ends and that of the nurse practitioner starts. In this paper the author discusses the problems associated with this confusion, most notably practice variation and argues that there is a need for guidelines for advanced nursing practice and rigorous evaluation. PMID- 8716494 TI - Floor patterns limit wandering of people with Alzheimer's. AB - The safety of people with dementia who walk into potentially dangerous areas, or away from areas where they can be monitored, is a problem in psychiatric settings for older people. Current methods of controlling these patients usually involve physical interventions such as restraint, locked ward doors and sometimes medication. Such methods can be costly, counter-productive and take carers away from other work. It has been observed that patients with Alzheimer's disease appear to be affected by certain visual stimuli that do not affect the wandering of non-demented people or staff members. Such visual stimuli can be a simple two dimensional grid pattern, which patients with Alzheimer's disease tend to perceive as a barrier. This effect is not usually seen in other types of dementia. Capitalising on this observation, black insulation tapes in two different grid configurations were laid out in an attempt to prevent these patients wandering through exit doors. Ten patients with differential diagnosis of dementia participated in this study. It was found that the use of a horizontal grid reduced exit door contact by up to 97% for four of these patients with a diagnosis of Alzheimer's disease. As had been predicted, the grid was less effective for patients with other types of dementia. Advantages and inherent dangers in this approach to managing hazardous walking by Alzheimer's patients are discussed. PMID- 8716496 TI - Vitamins: how much do we need? PMID- 8716498 TI - Reflections on the adult development and aging of the baby boomers. PMID- 8716499 TI - Poisonings in the elderly. PMID- 8716500 TI - How to maintain your ANCC certification. PMID- 8716501 TI - Library services for Oklahoma nurses. PMID- 8716502 TI - A day in the life of a public health nurse. PMID- 8716504 TI - 1995 Fuld Fellowship. Transforming nursing's vision: history, holism and the health care system. PMID- 8716505 TI - Position statement. Restructuring, work redesign, and the job and career security of registered nurses. American Nurses Association. AB - The American Nurses Association (ANA) is committed to safeguarding the public, protecting and advancing the careers of professional nurses, supporting individual and collective efforts by registered nurses to protect their clients and enhancing the professional development and job security of registered nurses. As the nation's health care system is restructured, ANA is actively engaged in initiatives to strengthen the economic and general welfare of registered nurses, the safety and care for the public, and, in partnership with the state nurses associations (SNAs), oppose efforts to replace registered nurses with inappropriate substitutes. PMID- 8716507 TI - [Will better hospital care be achieved in the near future?]. PMID- 8716506 TI - Position statement. The right to accept or reject an assignment. American Nurses Association. AB - The American Nurses Association (ANA) believes that nurses should reject assignment that puts patients or themselves in serious, immediate jeopardy. ANA supports the nurses obligation to reject an assignment in these situations even where there is not a specific legal protection for rejecting such an assignment. The professional obligations to the nurse to safeguard clients are grounded in the ethical norms of the profession, the Standards of Clinical Nursing Practice and state nurse practice acts. PMID- 8716509 TI - [Nursing courses in Germany. 2. The demand is great]. PMID- 8716508 TI - [International Hartmann-Wound Congress in Stuttgart. Physicians and nurses must become partners]. PMID- 8716510 TI - [Publicity work for administrators of old age homes. How to inform the press]. PMID- 8716511 TI - [Self-care for nurses. 2. A boon for overworked legs]. PMID- 8716512 TI - [Fixed work times and room care. I. A new model makes life easier for nurses]. PMID- 8716513 TI - [Holistic promoting process nursing in the ambulatory area. A model proves itself in empirical testing]. PMID- 8716503 TI - Pediatric AIDS amd camp nursing: a shared experience. PMID- 8716514 TI - [Functional urinary incontinence. Holistic promoting process nursing increases independence]. PMID- 8716515 TI - [Professional English for nurses: a new subject should become standard]. PMID- 8716516 TI - [A challenge for continuing education: a key to qualification]. PMID- 8716517 TI - [Federal regulation on nursing fees. The future of hospital financing is still open]. PMID- 8716518 TI - [Private practice infections and hospital infections: patients are suing for compensation]. PMID- 8716519 TI - [Second step of nursing insurance. Does an unconstitutional part-insurance ruin the nursing homes?]. PMID- 8716520 TI - [Dehospitalization. Where is the advantage for geriatric-psychiatric long-term patients?]. PMID- 8716521 TI - [Against stress and insomnia: "alternative relaxation" with mental systems]. PMID- 8716522 TI - [Critical considerations on the handing over of patients from one nurse to another at the bedside]. PMID- 8716523 TI - Neonatal individualized predictive pathway (NIPP): a discharge planning tool for parents. AB - The development of clinical pathways has paralleled the national movement to control health care dollars. This movement is particularly intense in states with extensive managed care plans, such as Arizona. Early infant discharge with minimal home nursing support is a reality that forces parents to anticipate their discharge needs well in advance of the actual discharge date. The 59-bed NICU at Phoenix Children's Hospital uses neonatal individualized predictive pathways (NIPPs) as a discharge planning tool with parents of premature infants less than 32-weeks gestation. The NIPPs were developed after an extensive retrospective record review identified time frames for a variety of medical therapies and clinical milestones. Parent and nursing staff expectations are specified on a weekly basis opposite the medical therapies. This article discusses the positive parental response to the NIPP and identifies future areas for development. PMID- 8716525 TI - The effects of swaddling versus standard positioning on neuromuscular development in very low birth weight infants. AB - A randomized control design was used to compare the effect of swaddling to standard positioning on neuromuscular development in very low birth weight (VLBW) infants (< 1,250 gm). The outcome of neuromuscular development was measured at 34 weeks postconceptional age using the Morgan Neonatal Neurobehavioral Exam (MNNE). The sample included 50 infants who met criteria for birth weight, age and who were classified as appropriate for gestational age. Exclusion criteria were also used. The infants were randomly assigned to the experimental group or the comparison group. Data analysis included descriptive and inferential statistical techniques. The results demonstrated that swaddled infants had higher total scores on the MNNE as compared to infants with standard positioning. Swaddling appears to be a positioning technique that can enhance neuromuscular development of the very low birth weight infant. PMID- 8716524 TI - Evaluation of oxygen delivery with the use of nasopharyngeal catheters and nasal cannulas. AB - The purpose of this pilot study was to examine the differences between nasopharyngeal catheter and nasal cannula oxygen delivery systems in infants. Eleven subjects requiring supplemental oxygen were enrolled in the study. All infants received both therapies at three separate gas flow rates (0.5, 0.3, and 0.2 liters per minute). Infants had higher heart rates when receiving nasal cannula oxygen than when receiving nasopharyngeal catheter oxygen (p < .05). There were no significant differences between the two therapies when comparing pulmonary mechanics, hypopharyngeal oxygen concentrations, and respiratory rates. However, when oxygen was delivered at the higher flow (0.5 liters), heart rate, tidal volume, and oxygen concentration were higher, and respiratory rates were decreased for both methods (p < .05). Because minimal differences were found between these two methods, either may be an appropriate choice for delivering supplemental oxygen. PMID- 8716526 TI - Validating assessment of neonatal jaundice with transcutaneous bilirubin measurement. AB - In response to current trends in maternity care, nurses are assuming a major role in providing quality care for new mothers and their babies postdischarge through follow-up home care. Jaundice has been identified as the most commonly encountered cause of morbidity among healthy infants during the first week of life, and nurses must possess accurate neonatal assessment skills. Unlike nurses who work in hospital settings, those who provide home care do not have ready access to colleagues for consultation and validation of assessments. The result of unrefined assessment skills of nurses may be requests for unnecessary invasive procedures for infants and, consequently, monetary charges to patients for unnecessary lab work. A quasi-experimental design was used to determine whether the use of a jaundice meter to validate nurses' visual assessments would help them develop more accurate assessments of neonatal jaundice. Findings revealed high correlations between visual scale rankings of jaundice and jaundice meter readings (RS = .85, p < .01). The results of this pilot study indicate that the use of a jaundice meter may be a noninvasive as well as a cost-effective way to validate nurses' visual assessments of neonatal jaundice in the home. PMID- 8716527 TI - UACs not culprit. PMID- 8716528 TI - State boards considering competency exams for nurse practitioners. NANN Board of Directors. PMID- 8716529 TI - Contemplating baby-friendly in the NICU? PMID- 8716531 TI - Research utilization: modifying the NICU light environment. PMID- 8716530 TI - The Canada-New Zealand connection. PMID- 8716532 TI - Kangaroo care videotape. PMID- 8716533 TI - Bronchopulmonary dysplasia. PMID- 8716535 TI - Learning disabilities nursing: developing new horizons. PMID- 8716534 TI - Incorporating physiologic principles into neonatal education. PMID- 8716536 TI - NVQs in nursing, midwifery and health visiting: a question of assessment and learning? AB - The nursing, midwifery and health visiting professions are faced with the question of whether there should be development of higher level NVQs within these professions. However, an examination of the assessment process in NVQs finds it to be bureaucratic, reductionist and fragmented in nature and unlikely to engender analysis and synthesis. Educational research has revealed that assessment has a powerful effect on learning. When applying the findings of this research to NVQs, the assessment in NVQs is found to have the potential to fragment and damage the learning striven for. It is unlikely to promote learning or maximize candidates potential capabilities in terms of development. The professions in Australia have developed an holistic approach to competence. The assessment process used there is in keeping with the assessment process in professional nursing, midwifery and health visiting education in England. It is important that the statutory bodies and the nursing, midwifery and health visiting professions consider the assessment process in NVQs, its potential effect on learning and the development of the professional practitioner within the context of future collaboration regarding the development of higher level NVQs. PMID- 8716537 TI - An exploration of the psychosocial perspectives on educational/developmental opportunities for enrolled nurses: the forgotten species? AB - The provision of further education opportunities is generally known to be limited for enrolled nurses. With the phasing out of the enrolled nurse qualification, it appears that many of them have to compete for limited places on conversion courses. Such a situation which has been placed on them appears to create problems in their lives. This study was undertaken with a randomized sample of 30 enrolled nurses to find out what the psychosocial problems may be for them who have not yet been able to convert to registered nurse status. The findings reveal a sense of betrayal, frustration, anger and helplessness at being coerced into getting onto conversion courses. That is compounded by the fact that places on such courses are extremely limited. Superimposed on those, fears were also expressed for their jobs. That was particularly so in the case of nurses from the area of learning disabilities. Although there are suggestions that they need to get onto conversion courses, there is a sense of disillusionment as places on the courses are extremely limited. Whilst the intention to convert may be present, many feel prevented from doing so because of their family commitments. In many instances, those commitments were non-existent when they first embarked on their nurse education/training. At the very least, the findings suggest a moral responsibility on the part of the relevant authorities to undertake a coordinated effort to help this group of nurses. After all it may be suggested that their predicament is not of their own making. PMID- 8716538 TI - Towards a philosophy of continuing professional education in nursing, midwifery and health visiting. AB - This paper outlines a justification for developing and articulating a belief system which encompasses continuing professional education and the theory and practice of nursing. It describes the nature and rationale for such a belief system, distinguishing it from the pursuit of an absolute or philosophy. Nevertheless, it is suggested that such a belief system owes and contributes much to both a philosophy of education and of nursing. PMID- 8716539 TI - Learning through outdoor activities as part of a P2000 mental health branch programme. AB - 'Outdoor activity' training was introduced into a P2000 Mental Health Branch Programme as a means of developing a range of personal and interpersonal qualities and skills. In this article, the authors outline the process of setting up this venture, identify aspects of the curriculum relevant to outdoor activities, giving examples of some of the activities and make links to nursing practice. Finally, an evaluation of this project is offered which suggests positive outcomes as a result of these teaching and learning strategies. PMID- 8716540 TI - Fast-tracking in baccalaureate nursing education: one Canadian response to students with advanced standing. AB - Accelerated nursing programmes for non-nursing college graduates were developed primarily to address a shortage of nurses in the USA. This article describes a unique Canadian programme which has evolved in response to demand from an increasingly older and well-qualified applicant pool for recognition of their relevant previous educational experience. In an 8-week period in the summer of 1993, 18 students completed the requirements for Year I of the 4-year baccalaureate nursing programme. When compared to their generic counterparts, these 'fast-trackers' were found to be older, more likely to be part-time and, at the end of Year 2 were academically equivalent. Students in the programme were highly enthusiastic, citing the benefits of saving a year of student costs and of entering the profession a year earlier. Faculty involved with the programme were positive about the benefits for students, although a drawback for full-time faculty was the impact of a heavy summer teaching load on other scholarly activities. Because of the programme's success, consideration is now being given to extending the fast-track option beyond Year I. PMID- 8716541 TI - The curriculum evolution and Project 2000: a critical examination. AB - The changes in nursing education underway in the UK and the USA are designed to educate a new type of practitioner. These new practitioners will be emancipated, critically reflective, creative and autonomous in their practice. This paper looks at the progress of the revolution, with particular reference to the UK, by critically examining the literature arising out of it. The implications for nursing education and educators are examined. PMID- 8716542 TI - Objectively assessing nursing practices: a curricular development. AB - In preparation for the changing needs of undergraduate nursing students undertaking a Project 2000 degree, it was necessary to rethink the nursing skills programme. After studying the literature a nursing skills laboratory was designed which provided both an institutional and a domestic setting. A progressive programme was developed, to help the students learn nursing practices, which was based on the Objective Structured Clinical Evaluation (OSCE). A small pilot study was set up using second and third-year students from the traditional nursing studies degree. A number of stations were set up comprising various nursing scenarios. The students who were being assessed rotated through these. Other students acted as patients, examiners and some volunteered to be novices being taught by the more senior students. A set of marking criteria was drawn up for each station to enable each student to be assessed objectively. One of the stations was filmed to provide the students with personal feedback. By the end of the session the students had rotated through each of the stations and received the marked criteria as feedback. At the end of the session a focused group interview took place with all the students and the two lecturers involved in setting up the project. Students were positive and felt the process had potential for future development as a means of integration and consolidation of skills prior to clinical experience. The early introduction of filming to the programme was though to be of benefit by reducing stress levels through regular use. Students felt that the role of teaching the 'novice' helped them focus on their knowledge and performance. This process is resource intensive in human and non human terms but enables small groups of students to learn in a realistic but safe, non-threatening environment and encourages them to take responsibility for their own learning. PMID- 8716543 TI - The 'art' of reflection: using drawing to uncover knowledge development in student nurses. AB - This paper describes how a nursing programme that is underpinned by the ideology of critical social theory used the medium of drawing to allow students to express learning that occurred on their clinical placement. The drawings were then examined to identify the different forms of knowing that the students depicted. It was found that students could demonstrate an awareness of different forms of knowledge. PMID- 8716544 TI - The need for change in nurse education: a literature review. AB - The implementation of the Project 2000 diploma course in 1989 meant that nurse education was moving rapidly from an apprenticeship system of preparation to an education led system in which students have student status. There is nothing new in the idea of student status for nurse learners; it had been suggested over 40 years before it actually came to fruition. This paper critically analyses the literature and looks at the reasons for the need for change in nurse education. An examination is made of the reasons why this change was not implemented earlier. The Project 2000 proposals are then explored with the benefit of hindsight, and the conclusion is reached that there is need for ongoing research to evaluate the impact of the course not that it is the established method of preparation for nurse learners. PMID- 8716546 TI - Gods, daemons and banshees on the journey to the magic scroll: the use of myth as a framework for reflective practice in nurse education. AB - This paper reports the use of a mythic journey scenario within a student nurse groupwork session. This approach was introduced as a means to facilitate personal and interpersonal sharing and self-knowledge. These processes are held as important for the development of critical reflective process as a cornerstone of contemporary nursing practice. The use of myth invoked a coherent and supportive student environment within which previously reticent students appeared confident enough to contribute and explore their educational experience in nursing. The mythic allegoric process also identified some poignant themes and enemies, common across the student body. Monsters no less! PMID- 8716547 TI - Preparing students for health promotion: the challenge for Australian nursing in the 90s. AB - Health promotion is increasingly recognized as a role to be taken up by all health workers, and nurses, in particular, have been recognized for their potential contribution in this area. Nurse educators around Australia are currently grappling with how best to reorient nurses towards health promotion, and how to prepare nursing students for a health promotion role. In this discussion paper, one university's preparation of their undergraduate students will be examined, as a means of stimulating further debate about how health promotion can be effectively incorporated into undergraduate nursing education. The Health Education and Promotion unit at University of New England, Armidale is offered in the third year of the Bachelor of Nursing. It incorporates traditional teaching strategies and a specific health promotion practicum. The practicum is vital in developing students' health promotion skills and helping them to integrate their theoretical learning into practice. It is argued that education in health promotion is vital if nurses are to prepare themselves for the challenge of promoting the health of their communities. PMID- 8716545 TI - Numeracy needs of the beginning registered nurse. AB - The literature indicates: Nursing students demonstrate serious numeracy skills deficits. Consensus that such skills are necessary, particularly for medication administration. Diagnostic testing and remediation are desirable, or basic mathematics is pre-requisite for entry into nursing courses. Calculation skills mastered at one point in time diminish if ongoing practise is not maintained. The frequency with which nurses perform drug calculation in the clinical setting is decreasing. This study aimed to identify and analyze numeracy skills required by beginning registered nurses to facilitate development of numeracy skills by nursing students. A convenience sample of metropolitan and rural health care facilities in New South Wales (NSW) was selected. Within each facility an area where nursing graduates customarily work was selected. Numeracy input required to complete patients' charts was identified. Additional counting, measuring or computation the nurses performed was also identified. Except for paediatric areas, the frequency with which a nurse has to perform other than a simple calculation in order to administer a medication, or calculate a per minute drip rate is low. However, the necessity to calculate drug doses does exist. Creative ideas are needed to enable nurses to do this accurately. Three general approaches are suggested. A number of recommendations are presented. PMID- 8716548 TI - Response to 'Midwives in current debate and in the future'. PMID- 8716549 TI - Midwifery--the turning point? AB - This paper endeavours to respond to suggestions about the 'fate' of midwifery in Australia and to provide stimulus for further discourse and analysis about where and how midwifery is going and growing, now and in the future. The tone is set as one of optimism and self-determination, as midwifery evolves within its history in Australia and finds and affirms, its own way from nursing and medico obstetrics its own genre with women, and its own unique heritage. It is argued that only with a self-determined commitment to the integrity of midwifery and of women (seeing the two as inseparable if we truly are with women). Will we as midwives be able to transcend notions of dependence, control, submission, and competitiveness among others. The writer has specifically chosen to present this paper without references to make clear the content of the response without the camouflage of other writers. It is hoped this paper may provoke other midwives to examine and debate their perspectives and ideas. PMID- 8716550 TI - Improving midwifery practice through clinical pathways. PMID- 8716551 TI - More than just pain relief: a study of postpartum, post-caesarean pain management. PMID- 8716552 TI - A theory of caring for midwifery. AB - It is time that Australian midwives developed and published their own theories and models for midwifery. This paper proposes a practical model for midwifery based on caring concepts. The needs of midwifery clients have been analysed and supported by various authors and four concepts of caring have been selected and applied to these needs to bring about a positive birth experience. Philosophical assumptions have been made to form the grounding for the theory, the core of which is a caring midwifery interaction. PMID- 8716553 TI - Changing patterns of care of pregnant women: the independent midwife. PMID- 8716554 TI - Focusing on men. PMID- 8716555 TI - Gloves: the last resort. PMID- 8716556 TI - Empowering clients: evaluating employee needs. PMID- 8716557 TI - Time management: getting the most out of the day. PMID- 8716558 TI - Professional misconduct update. PMID- 8716559 TI - Preventing osteoporosis: an independent study (continuing education credit). PMID- 8716560 TI - Empowerment as a leadership theory. AB - Leadership of today is quite different from that of Caesar, Plato, the ancient Greeks, or the ancient Egyptians. Today's leaders utilize power-sharing, trust, team-building, and empowerment. Methods to achieve empowerment include educating, leading, structuring, providing, mentoring, and actualizing. Empowerment requires change in leadership methods to be effective. One must be able to identify change and work with others for change. Through the leadership style of empowerment, vision provides a shared set of values with a customer focus. Successful organizations which practice empowerment delegate and expect people to initiate, create and to feel ownership. Nurses need to be involved in their organizations and utilize empowerment leadership to be successful leaders of the future. With empowered employees, the organization, the customer, and the employee are all winners. PMID- 8716561 TI - Mandatory drug testing in the workplace. AB - Although employees may object to mandatory drug testing due to concerns for an invasion of their privacy, employers are responsible for maintaining the public's safety. Education about substance abuse, testing, and treatment should be a part of every employer's anti-drug program. Costs for testing can range from $20 to $200; however, an employer, such as a hospital, that fails to test for substance abuse could face increased liability for an injury to a patient if a drug impaired employee caused it. PMID- 8716562 TI - Keeping competency up when patient numbers are down. PMID- 8716563 TI - Forty years of nursing: a perspective. PMID- 8716564 TI - Preparing camera-ready figures. AB - Today, authors can develop figures to illustrate their articles or books because of new computer graphics programs. However, some publication standards must be met in order for publishers to use the illustrations. This article describes current standards that will help authors develop high-quality figures. PMID- 8716565 TI - Write it now, or never! AB - Writers need knowledge and credentials to write, but one key to successful writing is to start writing now. Often the only difference between a published writer and a unpublished writer is simply that the published writer did the writing. PMID- 8716566 TI - Publication of a thesis: the relationship between graduate student and thesis advisor. AB - Identifying the roles the graduate student and the thesis advisor will undertake when preparing the thesis helps when the thesis is published. Such issues as bylines and copyrights should be agreed upon before embarking on the thesis work. The author, an Associate Professor in the nursing department at a state university, clarifies the roles of the graduate student and thesis advisor. PMID- 8716567 TI - Putting your toe in the water. PMID- 8716569 TI - Inventory management. PMID- 8716568 TI - Outcome-based practice: convince me! PMID- 8716570 TI - The grass may be greener in another pasture: moving outside the walls. PMID- 8716571 TI - New opportunities: beyond the walls of the hospital. PMID- 8716572 TI - Nurse leadership competencies for the 21st century. AB - Nurse managers and administrators must understand the competencies necessary to be successful in a changing health care environment. Critical leadership competencies have a significant impact on patient outcomes, cultivating and environment for continuous learning, interdepartmental and intradepartmental customer relationships within and across agencies, use of resources, strategic planning, and compliance with regulatory requirements. This article presents a review of literature and expert thought related to critical administrative and leadership competencies. The skills and competencies the experts discuss are universal, suggesting that the work venue is irrelevant to nurse leadership behavior, which translates across settings. PMID- 8716573 TI - Measuring competencies: development of the administrator competency survey. AB - Competency assessments is critical for nurse managers who must move between and among health care settings. This article reports development of a survey tool to measure administrative competencies to facilitate career movement based on competencies versus traditional structure of education and experience. The study was designed to determine specific competencies that were deductively derived and considered essential for successful leadership in the year 2000. These included eight domains: quality improvement, shared visions, global approach, serving community, mastering change, team learning, managing customer relationships, and managing a diverse workforce. The tool was used to measure nurse administrator competency in a select group. Results identified expertise and targeted priorities for training and improvement. PMID- 8716574 TI - Nurse manager learning styles in a learning environment. AB - This study focused on the development of guidelines for a training program for nurse managers in a newly formed integrated delivery system. Subjects' learning styles as defined in Kolb's Experimental Learning Theory were identified and graphically plotted in the appropriate learning environment or domain. Necessary training methods derived from Fry's Learning Environment table were applied to scores. A logical method is presented linking previous nursing student learning style studies with a cognitive schema experiment conducted with nursing novices and experts. The hypothesis that a subtle transition of learning style does occur from the concrete-divergent to the abstract-convergent as the nurse manager gains in education and moves through the management experience was supported. PMID- 8716575 TI - Nursing research in the hospital and beyond. AB - Outside is conceptualized as (1) settings in a larger system of care and (2) ways in which nursing transcends roles and relationships traditionally found in hospitals. The hospitals is one of many settings in the continuum of care. Although episodic hospitalization may be required, the health experience involves interface with sites outside the hospital. Nurses transcend the conventional expectation that they are implementors rather than orginators and that they are interdisciplinary team members rather than team leaders. A framework is presented that is compatible with this conceptualization in terms of nursing research, and scenarios illustrate strategies for nursing research using integration and maximization of existing resources. PMID- 8716577 TI - Allegiance to.....?? PMID- 8716576 TI - Psychiatric clinical nurse specialists as intensive case managers for the seriously mentally ill. AB - The government estimates the number of seriously mentally ill as high as three million, with annual cost for the care of this population approaching billions of dollars. This article describes a program for the seriously mentally ill in the community using psychiatric clinical nurse specialists as intensive case managers. Interventions regarding disease relapse symptomology and early recognition of medication side effects assist clients to seek earlier intervention. Desired outcomes include earlier hospitalizations at less serious stages, reduced length of stay, higher functioning, improved self-care skills, and improvement in obtaining and receiving medical service. With clinical nurse specialists in case management roles, clients receive treatment at more appropriate and less costly service levels. PMID- 8716578 TI - Preventing and controlling antimicrobial-resistant infections in hospitalized adults. PMID- 8716579 TI - Nutrition screening: an essential assessment parameter. AB - Nutrition plays a significant role in preventing the incidence and prevalence of disease and more importantly, promoting health. Nutrition screening is an important tool used by nurses and dietitians to identify and assess clients at risk for nutritional compromise. PMID- 8716581 TI - Nursing management of adults who have cystic fibrosis. AB - Cystic fibrosis has ceased to be a terminal illness for children and has become a chronic disease affecting both children and adults. Life expectancy, already over 29 years, is increasing with new therapeutic interventions. Pathophysiology, assessment, and treatment information is presented here to assist adult health nurses in caring for patients with cystic fibrosis. PMID- 8716580 TI - Evaluating physical functional outcomes: one category of the NOC system. AB - All professional health care disciplines are now being held accountable to demonstrate the outcomes they produce. It is imperative that nurses demonstrate the difference they make in patient care to justify the cost of professional nursing. Standardized language is needed to aggregate data and report information on nursing interventions and associated outcomes. The Nursing Outcomes Classification (NOC) system is the first comprehensive classification of nursing sensitive patient outcomes. In this review, the development of this classification system with specific results related to physical functional status and implications of NOC for nursing practice are discussed. PMID- 8716582 TI - Reducing post critical care infection. AB - Infection is a significant concern for seriously ill patients transferred to medical or surgical units following critical care. Understanding the factors that contribute to pulmonary, urinary, wound, and vascular access infections enables nurses to effectively intervene to lessen this risk. Key elements in reducing susceptibility are discussed. PMID- 8716584 TI - Motivation and the "E" word (exercise). PMID- 8716583 TI - Anticancer drugs for noncancer diseases. AB - An increasing body of evidence supports the use of anticancer drugs to control symptoms and slow the progress of immune-mediated disorders. Drugs such as nitrogen mustard, azathioprine, and methotrexate, among others, are now being used to treat several types of immune-mediated disorders. However, the use of anticancer drugs for noncancer diseases differs significantly from their traditional use in treating cancer and has important implications for nursing care. PMID- 8716585 TI - The case management role in utilization review. PMID- 8716586 TI - Psychotropic medications and the elderly. PMID- 8716587 TI - Peristomal pyoderma gangrenosum: review and case study. PMID- 8716589 TI - A progressive management tool. PMID- 8716588 TI - Sexual harassment: a practical guide for nurses. PMID- 8716591 TI - The Internet and nursing: a vital link. PMID- 8716590 TI - Deep vein thrombosis and pulmonary embolism prevention: what role do nurses play? PMID- 8716592 TI - Ethics and managed care: beyond helplessness. PMID- 8716594 TI - Beyond mechanical nursing care. PMID- 8716593 TI - The ethics of valuing nursing. PMID- 8716596 TI - Broad-based research agenda address regulatory issues faced by member boards. PMID- 8716595 TI - The greening of horticultural therapy. Interview by M Elabdi. PMID- 8716598 TI - Overview of the Computerized Clinical Simulation Testing (CST) project. PMID- 8716597 TI - National Council's Special Services Division (SSD) takes off. Nursing educator workshops and certification examination for practical and vocational nurses in long-term care first on the agenda. PMID- 8716601 TI - NCLEX-PN test plan to change: new test plan to be implemented in October 1996. PMID- 8716599 TI - Why disciplinary databanks? Why the National Practitioner Data Bank (NPDB)? PMID- 8716600 TI - Questions and answers about becoming an NCLEX item writer. PMID- 8716602 TI - Update on the Nurse Information System (NIS). PMID- 8716603 TI - [Nurses are angels to patients]. PMID- 8716604 TI - [Delegation of authority based on review of decision making process. Need for delegation of authority and successful administration of organization]. PMID- 8716606 TI - [Delegation of authority based on review of decision making process. Delegation of authority to nurse supervisors has served as incentives for staff nurses]. PMID- 8716605 TI - [Delegation of authority based on review of decision making process. Reform of report system of the nursing department]. PMID- 8716607 TI - [Delegation of authority based on review of decision making process. More authority delegated to deputy directors of nursing has proven its merits and demerits]. PMID- 8716608 TI - [Delegation of authority based on review of decision making process. Delegating authority facilitates professional development of individuals involved]. PMID- 8716609 TI - [Delegation of authority based on review of decision making process. Nursing reform at Boston's Beth Israel Hospitals]. PMID- 8716610 TI - [Life of elderly patients at Farnham Hospital in England]. PMID- 8716612 TI - [Report of a hospital director of nursing--function of top management in a hospital nursing service]. PMID- 8716613 TI - [History of Japanese nursing that I lived: the Ministry of Health and Welfare's health policy to combat TB in the 1930s]. PMID- 8716611 TI - [National nursing policy. Interim report on newly proposed caregiving system for the elderly]. PMID- 8716614 TI - [Medical topics. Gunshot wounds]. PMID- 8716617 TI - [Liberation of medical service--who is responsible for closure of medical information and why?]. PMID- 8716616 TI - [Court case of nursing malpractice--failure to practice mouth-to-mouth resuscitation to an asthmatic child with apnea and heart failure follow i.v. treatment for an asthma attack]. PMID- 8716615 TI - [Specialist nurses in private practice--activities of the Continence Center, Inc. for five years since its establishment]. PMID- 8716618 TI - [Creation of clinical nurse specialists to work in ICU--who should employ them?]. PMID- 8716619 TI - [Summary of report on the 1993 survey of actual status of nursing personnel by the Japanese Nursing Association]. PMID- 8716620 TI - [Report of the Ministry of Health and Welfare's Working Party on Informed Consent]. PMID- 8716622 TI - SNAs raise nursing's profile during Nurses Week. PMID- 8716623 TI - Reviewing and reflecting the President's perspective: 1992-1996. PMID- 8716621 TI - ANA's landmark patient safety legislation debuts on Capitol Hill. PMID- 8716624 TI - Nursing skills ideal for entrepreneurship. As I see it. PMID- 8716625 TI - Something to celebrate. PMID- 8716627 TI - ANA's position on assisted suicide. PMID- 8716626 TI - ANA advises nurses not to participate in assisted suicide: RNs should consider implications of possible legalization. PMID- 8716629 TI - Tipper Gore urges nurses to advocate for mental health services in managed care. Interview by Virginia Trotter Betts. PMID- 8716628 TI - Nurses play key role in war on tobacco: new guideline supports nurses' consumer education efforts. PMID- 8716630 TI - Consumers echo RN concerns about use of unlicensed aides. PMID- 8716631 TI - Today's home health nurses rely on new technology. PMID- 8716632 TI - North Carolina establishes first Council on Managed Care. PMID- 8716634 TI - ANA Centennial. Good old days? PMID- 8716635 TI - Purchasing health care in the UK. PMID- 8716633 TI - ANA's Centennial: the birth of a professional organization. PMID- 8716636 TI - Rudderless debates. PMID- 8716637 TI - Unlock the leadership potential. PMID- 8716638 TI - Managing change. PMID- 8716640 TI - Leading to primary care. PMID- 8716639 TI - The politics of quality. PMID- 8716641 TI - Purchasing home care. PMID- 8716642 TI - The New World of commissioning. PMID- 8716643 TI - Leadership and nursing: Part Two: Styles of leadership. PMID- 8716644 TI - A leap in the dark. PMID- 8716645 TI - Effective purchasing: the nurse's role. PMID- 8716646 TI - [Experimental test of the bacteriostatic and bactericidal effect of six disinfectants for Pseudomonas aeruginosa]. AB - Minimal inhibitory concentration (MIC) and minimal bacteriocidal concentration (MBC) of six chosen disinfectants against pseudomonas aeruginosa were tested by the tubular liquid double dilution method. Results showed that 0.06% Chang Kou Jing and 1.5% Supertime are fairly good disinfectants against pseudomonas aeruginosa in infected wound surfaces, postoperative wound dressing, disinfection of surgeons' arms and hands before operation, sterilization of instruments and scrub medical workers' hands after contact with infected patients. The MIC and MBC of the former were 7.5 micrograms/ml and 30.0 micrograms/ml separately, while those of the latter were 11.7 micrograms/ml and 93 micrograms/ml respectively. PMID- 8716647 TI - [Discharge advice for carcinoma of the esophagus and cardiac patients]. PMID- 8716649 TI - [Care of infant with posterior fossa tumor during surgery]. PMID- 8716648 TI - [Observation of 41 patients with diabetes insipidus after microsurgical transsphenoidal pituitary adenomectomy]. PMID- 8716650 TI - [Care of hypertension of pregnancy syndrome during parturition]. PMID- 8716652 TI - [Care of fluid infusion with indwelling arteriovenous needle in burn patients]. PMID- 8716653 TI - [Effect of stereopsis intensification training of amblyopia]. PMID- 8716651 TI - [Posture feeding for the treatment of neonatal peculiar stomach torsion]. PMID- 8716654 TI - [Care of semicircular canal cryosurgery for Meniere's disease before and after surgery]. PMID- 8716656 TI - [Several methods beneficial to the memory of students in studying basic nursing]. PMID- 8716655 TI - [Experience of controlling the quality utilizing the "head nurse manual book"]. PMID- 8716657 TI - [Care of one severe hypertension of pregnancy patient complicated with abruptio placentae]. PMID- 8716658 TI - [Care during transcatheter plaque atherectomy of coronary artery]. PMID- 8716659 TI - [Study on current status of cardiac rehabilitation nursing for acute myocardial infarction patients in China]. PMID- 8716660 TI - [Investigation analysis of tumor patients requirements for education]. PMID- 8716661 TI - [Relationship between an aged mother and grown-up son left to themselves]. PMID- 8716662 TI - [Meeting with the Empress to explain the situation about the Hyogo earthquake disaster]. PMID- 8716663 TI - [Medical and health care of disaster victims--lessons learned from the great Hanshin earthquake. Care of victims at the closet neighboring life-saving emergency care center]. PMID- 8716664 TI - [Medical and health care of disaster victims--lessons learned from the great Hanshin earthquake. Nursing activities at a large general hospital built on artificial land with lifelines and the transportation system destroyed]. PMID- 8716665 TI - [Medical and health care of disaster victims--lessons learned from the great Hansin earthquake. Faculties and students of College of Health, Science and Art, Hyogo, acted to support nursing volunteer activities]. PMID- 8716666 TI - [Medical and health care of disaster victims--lessons learned from the great Hansin earthquake. Two weeks of a hospital located in the most severely stricken area]. PMID- 8716667 TI - [Medical and health care of disaster victims--lessons learned from the great Hansin earthquake. From my four days' experience as a volunteer pharmacist]. PMID- 8716668 TI - [Medical and health care of disaster victims--lessons learned from the great Hansin earthquake. Post-disaster visiting nursing service for home-bound patients]. PMID- 8716669 TI - [Medical and health care of disaster victims--lessons learned from the great Hansin earthquake. Helicopter transportation of earthquake victims]. PMID- 8716670 TI - [Medical and health care of disaster victims--lessons learned from the great Hansin earthquake. Mental health care for PTSD patients]. PMID- 8716671 TI - [Medical and health care of disaster victims--lessons learned from the great Hansin earthquake. Things I did as a nurse to help town people at an evacuation center]. PMID- 8716672 TI - [Medical treatment of patients of sarin intoxication]. PMID- 8716673 TI - [Report of a hospital director of nursing--hospital-based primary care at Saiseikai Hospital]. PMID- 8716674 TI - [History of Japanese nursing that I lived: development of community health nursing and visiting nursing service]. PMID- 8716675 TI - [Medical topics. Colostomy and stoma]. PMID- 8716676 TI - [Specialist nurses in private practice--continence adviser]. PMID- 8716677 TI - [Life of elderly patients at Farnham Hospital in England]. PMID- 8716678 TI - [Court case of nursing malpractice--death caused by disconnected artificial respirator]. PMID- 8716679 TI - [Liberation of medical service--provision of medical information to improve patient-doctor communication]. PMID- 8716680 TI - [Creation of clinical nurse specialist to work at ICU]. PMID- 8716681 TI - [U.S. nurses march on street to protest against RN reduction]. PMID- 8716682 TI - Designing new workers for tomorrow's world. Interview by Dina Leifer. PMID- 8716683 TI - Efficiency team confident of cutting NHS red tape. Interview by Pat Healy. PMID- 8716684 TI - Awareness is the cure. PMID- 8716685 TI - Bumph busters. PMID- 8716686 TI - Tackling taboos. PMID- 8716687 TI - The power of public knowledge. PMID- 8716688 TI - Living with depression. PMID- 8716689 TI - A holiday from home. PMID- 8716690 TI - A view from the cell. PMID- 8716691 TI - Using scientific knowledge. PMID- 8716692 TI - Treating leg ulcers: the latest techniques. AB - Several advances in the care and treatment of leg ulcers are currently being developed, including laser therapy, ultrasound and the use of hyperbaric oxygen. In this article, the author describes the theory behind each therapy and the current level of research knowledge on the effectiveness and use of the methods. PMID- 8716693 TI - Sun exposure and skin cancer development: nurses' attitudes. AB - To mark Sun Awareness Week next week, this article highlights the fact that the major contributory factor in the development of skin cancer is exposure to ultraviolet radiation, and nurses are ideally placed to promote care in the sun and raise awareness of moles. The aim of this study was to determine whether there are any gaps in nurses' knowledge about the prevention and early detection of skin cancer. A total of 142 nurses were questioned about their own attitudes towards sun exposure, sun protection and mole awareness. The study showed that the nurses surveyed have a responsible attitude towards avoiding sunburn and the need for adequate sun protection, particularly when abroad. However, the study also revealed that they do not fully appreciate the extent to which the sun can cause skin cancer and they lack understanding about the need to protect the skin from sunburn and avoid long term sun exposure in the UK. PMID- 8716694 TI - Using reflective diaries to assess students. AB - This article looks at the problems of using reflective diaries to conduct formative assessments of students' performance. The author highlights the importance of the proper definition of terms, and suggests there are strengths and limitations associated with attaching significance to retrospective, reflective self-assessment. PMID- 8716695 TI - AIDS, euthanasia and nursing. AB - Requests for euthanasia can pose a dilemma for nurses. Some of the arguments for and against euthanasia are presented here together with a summary of a study into the phenomenon of 'underground' euthanasia. These are considered in relation to the law and nursing ethics. PMID- 8716696 TI - Can I have an AIDS test? AB - Nurses in all areas will increasingly come into contact with people concerned about HIV infection. Although pre- and post-test counselling should be performed by those specially trained for it, sound knowledge about the physical, psychological, emotional and social effects of HIV will help nurses offer reassurance and advice. PMID- 8716697 TI - AIDS. Sharing knowledge. AB - There is a large and growing demand for HIV/AIDS services among residents of east London. Many of those infected or affected are refugees or asylum seekers who often fear statutory agencies. PMID- 8716698 TI - Department of Transportation 1995 regulations for alcohol testing: policy changes and compliance issues. PMID- 8716699 TI - From idea to publication: the secrets of publishing. AB - Publishing in lay and professional journals is essential to a practice profession such as nursing. The process of publishing an article begins with a focused idea, aimed at a specific audience, transmitted through a particular journal. To actually write the body of the article, authors must complete a library search, develop topic and supporting sentences within an organized outline, and create companion tables and figures to further explain the content. Responding appropriately to acceptance, rejection, or request for revision of the manuscript is key to eventually seeing the article in print. PMID- 8716700 TI - Coping with stress in a cross cultural setting: the case of Japanese and American employees of a Japanese plant in the United States. AB - The purpose of this pilot study was to compare the psychosocial distress of Japanese auto workers with that of the American employees working in a Japanese managed plant, to describe mediating factors related to their distress, and recommend interventions. Japanese and American workers and spouses responded to five questionnaires and two open ended questions about changes in health, function, and time, and needs, problems, and recommendations for stress reduction. Japanese men had higher General Health Questionnaire scores than American workers. Psychosocial distress was related to work self confidence and individual responsibility for Japanese men, with social support for Japanese women, and with individual responsibility for American men and women. Stress was related to communication problems and misunderstanding about the management practices and cultural norms. Culturally appropriate worksite and community interventions are discussed. PMID- 8716701 TI - Occupational health nurses and occupational hygiene: a study of South African nurses' attitudes. AB - The objectives of this study were to assess the current occupational hygiene practices of occupational health nurses and to assess their attitudes to the identification and initial quantification of workplace hazards. A questionnaire was mailed to all occupational health nurses registered with the South African Society of Occupational Health Nurses. Responses were obtained from 221 (53.7%). Responders and non-responders did not differ on key characteristics. Only 14 (6%) of the respondents performed occupational hygiene tasks as part of their routine work and only 31 (14%) volunteered hazard identification and quantification as tasks that would significantly improve practice. Nevertheless, when asked directly, 120 (54%) agreed that occupational hygiene fell into the ambit of occupational health nursing. Over 70% were positive about receiving theoretical and practical hygiene training. Constraints to greater hazard identification included limited time and resources and concern about intruding into the domains of other practitioners. Sufficient numbers of occupational health nurses were interested in identifying hazards in the workplace for training courses to be planned and offered now; however, restraints to practice need to be clarified and removed for these new skills to be used effectively. PMID- 8716702 TI - REACH: an alternative early return to work program. AB - One of the most frequently cited strategies for successful return to work post injury or illness--early return to work--is often an unattainable goal for many employees. Returning injured or ill employees to an existing program outside of the company (where the injury or illness occurred) is a means of overcoming the obstacles often associated with early return to work programs. A significant reduction in workers' compensation costs was demonstrated for employees participating in the early return to work program that placed ill or injured workers in positions at workshops for the developmentally disabled. This reduction in costs was demonstrated while paying these employees 100% of their wages rather than the lesser compensation amount. PMID- 8716703 TI - Health on the road: developing a program for international travelers. PMID- 8716704 TI - A computerized surveillance data management system using Epi Info. PMID- 8716705 TI - Mobile worksite programs. PMID- 8716706 TI - [Precautionary education against extracorporeal shock wave lithotripsy complication]. AB - The purpose of precautionary education against Extracorporeal Shock Wave Lithotripsy complication in urinary system is to help patients build knowledge, trust, and collaboration. The methods of the education include 1) group education -reading pamphlet which we specially provide and receiving regular information, 2) individual education--giving advice prior to treatment and something special to the patient's needs. Educational content include drinking water, physical adjustment and postural drainage. Three years clinical practice showed that precautionary education could reduce unnecessary complication after Extracorporeal Shock Wave Lithotripsy and it needs further attention in this field. PMID- 8716708 TI - [An investigation of post-secondary nursing continuing education]. AB - An investigation was made in 20 hospitals in Nanjing to the abilities of the post secondary college nursing graduates in 4 aspects: (1) nursing technical practice; (2) the numbers of head nurse employment; (3) nursing teaching; (4) nursing research and academic paper writing. Meanwhile a comparison was made between them and college graduates and baccalaureate graduates. Result showed that they were evidently better than baccalaureate graduates (P < 0.01) in nursing technical practice, head nurse employment, and nursing research and obviously different from college graduates (P < 0.01) in technical practice and nursing research. Post secondary continuing education in nursing, as one of the ways of higher nursing education yet adopted, has been playing an extremely important role in the improvement and development of nursing in China. PMID- 8716707 TI - [Study on the collecting method of human umbilical cord blood]. AB - The methods of cord blood collection was investigated in this study. The purpose of this study was to evaluate human umbilical cord blood as a alternative to bone marrow in the provision of transplantable progenitor cells for hematopoietic reconstitution. The results showed that using modified technique with a sterile and closed system, the volume of cord and placental blood would all exceed 120 ml. The average amount of blood collected was 132.2 +/- 12.13 ml and the highest was 158 ml. Cord blood culture for bacteria and fungi were negative. Blood clots were not present. And in the examination of each single cord blood, the contents of nucleated cells (NS) and mono-nucleated (MNS) were, respectively, 18.9 X 10 +/ 1.7 X 10, 8.2 X 10 +/- 0.8 X 10. The result suggested that the collection of placental blood with the modified technique in a sterile and closed system is a simple, safe and efficient procedure. The average amount of blood collected and hematopoietic cells would satisfy the needs of cord blood transplantation. Immediate cut of the umbilical cord followed by a prompt puncturing is critical for the volume and quality of the sample. PMID- 8716709 TI - [Behavior therapy of schizophrenia patients during convalescent period]. AB - 58 of schizophrenia patients at restoration stage in the hospital were studied. The ineffective adaptive behaviors were assessed and behavior therapy plans were made. The behavior therapy was provided in group or individually. The result indicated that patients' ineffective adaptive behaviors such as laziness, passive, withdrawal, role conflict, sexual dysfunction, ineffective adaptation in the society, and aggressive were improved a lot than those before receiving therapy (P < 0.01). PMID- 8716710 TI - [Using SpO2 meters to observe the circulation of replantation of severed fingers]. PMID- 8716711 TI - [Care of carcinoma of esophagus and heart after surgery complicated with pylorochesis]. PMID- 8716712 TI - [Care of epikeratophakia for aphakia in children]. PMID- 8716713 TI - [Care for the patient during hydroxyapatite ocular prosthesis implantation]. PMID- 8716714 TI - [Care of newborns with HIE treated by high-pressure oxygen]. PMID- 8716715 TI - [Application of encouraging theory in nursing management]. PMID- 8716716 TI - [Quality control of baccalaureate students practice of nursing process through systems management]. PMID- 8716717 TI - [Design of microcomputer management of testing questions in obstetrics and gynecology nursing]. PMID- 8716718 TI - [Experience in organizing English speech competition for nurses]. PMID- 8716719 TI - [Care of psychological demands of hospice patients]. PMID- 8716720 TI - [The present state of prevention and treatment of bedsores]. PMID- 8716721 TI - [What did nurses do and wear a hundred years ago in Japan?]. PMID- 8716722 TI - [Our expectations of the Japanese Nursing Association. Larger voice in formation of strategic policy]. PMID- 8716723 TI - [Our expectations of the Japanese Nursing Association. Nursing leadership required in introducing a public health insurance system]. PMID- 8716724 TI - [Our expectations of the Japanese Nursing Association. JNA should be made more friendly and close to its members at large]. PMID- 8716725 TI - [Our expectations of the Japanese Nursing Association. Association should be more responsive and sensitive to the needs of nurses practicing in clinical settings]. PMID- 8716726 TI - [Our expectations of the Japanese Nursing Association. Association's preparedeness for emergencies]. PMID- 8716727 TI - [Report of a hospital director of nursing--orientation for newly employed nurses at the start of the new fiscal year]. PMID- 8716728 TI - [History of Japanese nursing that I lived: prevention of TB and care of TB patients]. PMID- 8716729 TI - [Specialist nurses in private practice--studying incontinence care in the U.K]. PMID- 8716730 TI - [Life of elderly patients at Farnham Hospital in England]. PMID- 8716731 TI - [Court case of nursing malpractice--midwife's failure to monitor on the machine the fetal heart beat which caused cerebral palsy of the newborn]. PMID- 8716732 TI - [Liberation of medical service--publication by Medical Brain for health care consumers to become wise consumers]. PMID- 8716733 TI - [The nursing shortage and the 1990's: realities and remedies]. PMID- 8716734 TI - [On March 19 at the Connecticut Hospice of Brandford, Virginia Henderson died. With her dies a great nurse and a great woman]. PMID- 8716735 TI - [Continuing education to bring about change in the health services]. PMID- 8716737 TI - [Characteristics of the requests in primary care facilities]. PMID- 8716736 TI - [Posture treatment in hemiplegic patients]. PMID- 8716738 TI - [Sun baths, long-term enemies]. PMID- 8716739 TI - [The current development of the "working better" philosophy to reduce back strain]. PMID- 8716740 TI - [Nursing diagnosis: is there a common language?]. PMID- 8716741 TI - [The placental circulation system]. PMID- 8716742 TI - [Biomechanical aspects of the spine. Vertebral health]. PMID- 8716743 TI - [Students' health habits]. PMID- 8716744 TI - [Special beds. Types and indications]. PMID- 8716746 TI - The really useful guide to portfolios and profiles (repeat) (continuing education credit). PMID- 8716745 TI - [Liver transplantation. Control of infection]. PMID- 8716747 TI - [A longitudinal study of growth and development of low birth weight infants]. AB - In order to investigate the growth and development of low birth weight infant (LBWI), 132 low birth weight infants (BW < 2500g) born in 1983 living in Xi Hui district were followed up from birth to three years. And 49 full-term infants with normal birth weight were selected as controls. The weight, length, and head circumference of infants of two groups were measured periodically. Their development abilities were assessed by using Gesell Development Diagnosis. Results showed the weight, length, and head circumference of LBWI were obviously less than that of the controls. The developmental quotient (DQ) of LBWI were lower that that of controls, particularly in gross motor field, Therefore, it is important to reduce the rate of IBWI and to give scientific guidance on child care and early training for promoting growth and development of infants. PMID- 8716748 TI - [The effect of air purification on 10,000 degree clean room in supply rooms]. AB - This article introduced that purifying equipments were installed in the major operating departments of supply rooms in hospital, and that experimental tests were done on the packing department. In statil state, the air humidity and the number of bacteria in the room after purification was obviously lower than that before purification (P < 0.01). In trending conditions, the main technical index on 10,000 degree clean rooms were examined, all of the results were up to the national standards. PMID- 8716749 TI - [The effect of early chest physiotherapy on blood gas and circulatory function in old patients after thoracotomy]. AB - In this study, 15 old patients undergoing thoracotomy were selected for observing the influence of effective chest physiotherapy started early in postoperation on blood gas and circulatory function. Following ultrasonic aerosol inholation for 20 minutes, turning the body, and chest percussion in head-up were started at 6th hour after operation. The result showed that PaO2, SaO2 were significantly increased and PaCO2 were decreased at 30 minutes after treatment comparied with it before treatment. In other side, although the hemodynamics could maintain stabilization, cardiac arrhythmias were occured during treatment in three cases, whose ECG were abnormal in preopration. It concluded that effectively chest physiotherapy, started early in postoperation, could prevent postoperative hypoxemia in old thoracotomy patients, But, ECG monitoring is necessary during treatment in patients whose ECG was abnormal before operation. PMID- 8716750 TI - [Continuous infusion of morphine in use of pain relief after upper abdominal surgery]. AB - This study was designed as a prospective randomized comparative trial of continuous infusion of morphine (CIM) controlled by a micro-electric pump and traditional intermitted intramuscle injection of pethidine (IMP) to relieve pain after surgery. 40 patients submitted from upper abdominal surgery were divided into two groups, 20 patient receiving CIM and others using IMP. The amount of morphine used during the 24 hours was 0.5 mg/kg in CIM group. In IMP group, 20 patients were given standard intramuscular dosages of 1.5 mg/kg of pethidine, then, every 4-6 hours repeatedly within 24 hours after operation. An analgesic scale was observed by WHO's pain score from I to IV degree. The result showed that all patients of CIM group had good analgesic effect (I and II degree), however, 10% of the patients in IMP group had only analgesic effect of I and II degree during the first 24 hours after operation. Their respiratory frequency, heart rate, BP and SpO2 had no significant difference in two groups. It woule appear that CIM is an efficacious and safe method for providing postoperative pain relief. PMID- 8716752 TI - [Aviation care of burn patients]. PMID- 8716751 TI - [Care of amebic hepatic abscess]. PMID- 8716753 TI - [Care of children after selective posterior rhizotomy]. PMID- 8716754 TI - [Care of Blakemore tube for acute hemorrhage from esophageal varices in children]. PMID- 8716755 TI - [Pre and postoperative care of intramedullary epidermoid and dermoid cyst in children]. PMID- 8716756 TI - [Carrying out standardized management, improving nursing quality]. PMID- 8716757 TI - [Microcomputer management of chronic hepatitis B virus asymptomatic patients]. PMID- 8716759 TI - [Clinical observation of constipation due to deficiency of vital energy treated by massage and finger pressure methods]. PMID- 8716758 TI - [Investigation and strategy of nurses' continuing education in the Beijing area]. PMID- 8716760 TI - [Investigation of ratio of breast feeding of 1-4 month old infants of community area]. PMID- 8716761 TI - [Diagnosis and nursing care of postoperative fatigue]. PMID- 8716762 TI - [Systematic approach of holistic nursing and model unit establishment]. PMID- 8716764 TI - [Role analysis of nursing personnel and qualified caregivers. What are the unique activities of nursing personnel?]. PMID- 8716763 TI - [Sexless couples are increasing recently]. PMID- 8716765 TI - [Role analysis of nursing personnel and qualified caregivers. Role of nursing personnel and caregivers at home for elderly people with health problems]. PMID- 8716766 TI - [Role analysis of nursing personnel and qualified caregivers. Training programs to become qualified caregivers]. PMID- 8716767 TI - [Role analysis of nursing personnel and qualified caregivers. Division of work between nursing personnel and caregivers]. PMID- 8716768 TI - [Role analysis of nursing personnel and qualified caregivers. Proposed introduction of a caregiving insurance system and its effect on the nursing profession]. PMID- 8716769 TI - [Role analysis of nursing personnel and qualified caregivers. Caregiving insurance in Germany]. PMID- 8716770 TI - [National nursing policy. Report on informed consent by the Ministry's Special Committee]. PMID- 8716771 TI - [Report of hospital director of nursing--career development for nurses practicing in hospitals]. PMID- 8716772 TI - [History of Japanese nursing that I lived: history of the National Sanatorium Muramatsu Seirannson]. PMID- 8716773 TI - [Medical topics. Clinical work to prevent trauma]. PMID- 8716774 TI - [Tests and diagnosis of allergy]. PMID- 8716775 TI - [Specialist nurses in private practice--acceptance and non-acceptance of a continence nurse in our health system]. PMID- 8716776 TI - [Life of elderly patients at Farnham Hospital in England]. PMID- 8716777 TI - [Court case of nursing malpractice--a child receiving i.v. injection ina high pressure oxygen room suffered an air embolism which caused him a central nerve disorder]. PMID- 8716778 TI - [Liberation of medical service--health service networking]. PMID- 8716779 TI - [We learned so many things from the great Hanshin earthquake]. PMID- 8716780 TI - [Statement by Ministry of Health and Welfare's Committee. Improvement of educational programs of medical and health professions]. PMID- 8716781 TI - [International Confederation of Midwives' Asia Pacific Regional Conference held in Saitama, Japan]. PMID- 8716783 TI - Nursing as a career. PMID- 8716782 TI - Advanced practice clinics redefine the primary care provider. PMID- 8716784 TI - Managed care and changing nurse practice. PMID- 8716785 TI - Evolving roles for advanced practice nurses. PMID- 8716786 TI - A time for plain talking. PMID- 8716788 TI - Be wise with medicines. PMID- 8716787 TI - Healing the pain of Port Arthur. PMID- 8716789 TI - Consumers: aware and empowered. PMID- 8716790 TI - Competency standards for the advanced nurse clinician--have your say. PMID- 8716791 TI - Let us pray. PMID- 8716792 TI - Follow the leader: mentoring and health care. PMID- 8716793 TI - The role of a children's hospice. PMID- 8716794 TI - 'Resus' skills for parents. PMID- 8716795 TI - Jejunal biopsy--seeking an alternative. PMID- 8716796 TI - Expanding the boundaries of care. PMID- 8716797 TI - Parents as partners in care. PMID- 8716798 TI - Paediatric life support. PMID- 8716799 TI - A brief mental health outcome scale: relationships between scale scores and diagnostic/sociodemographic variables in the long-term mentally ill. AB - The Global Assessment of Functioning (GAF) scale is a quick and simple measure of overall psychological disturbance. A recent study by our group reported the reliability and validity of the scale in a population of severely mentally ill people. The current study extends the analysis of this population to consider a number of questions, i.e. what are the associations for this group between variations in GAF scores and sociodemographic variables? Are lower scores associated with particular diagnoses for the group assessed? What are the characteristics of those patients whose scores were most variable? What were the completion rates for use of the scale? No associations were observed between GAF score and the sociodemographic variables used in this study. Lower GAF scores were associated with psychotic diagnoses. Those with most varying scores were, in the main, white European non-immigrant males with a diagnosis of schizophrenia. Completion rates for the study were reasonable at 73.8%. This study involved the repeated assessment of a large number of clients by a changing multidisciplinary team. Inspite of the extended nature of the study, completion rates remained reasonable, suggesting that the scale was acceptable for routine clinical use. The GAF in itself, however, appears to be unsuitable for making individual treatment decision but could be used as a first step before using more sophisticated scales. PMID- 8716800 TI - Reconciling scientific and religious discourse about madness during the age of reason: lessons for today? AB - This paper argues that the secularization of madness, during the seventeenth and eighteenth centuries, occurred as a consequence of cultural change that accompanied the social upheavals of the age. In examining the reconciliation of competing explanations for madness, from theological and empirical viewpoints, it is suggested that these paradigms were never totally separated and argued that developments during this period were a consequence of continual interaction and dialogue between these contrasting views. Furthermore, it is suggested that an understanding of these changing times can illuminate present debates surrounding mental illness. PMID- 8716801 TI - Taking the pith out of reality: a reflexive methodology for psychiatric nursing research. AB - Positivist knowledge, although well respected, is only one kind of knowing. Positivist methodology has made a partial contribution to knowledge of psychiatric nursing practice. Knowledge that relates directly to practice is the most sought after by practitioners themselves. Practitioners are in the best position to undertake 'insider' research that allows the process, rather than the outcome, of practice to be explored. A social constructionist approach is congruent with an insider research position. Social constructionism entails more tolerance concerning how we know our world. However, this tolerance breeds difficulty in evaluating research findings. Overall, knowing from practice is problematic. Tacit knowledge acquired through professional socialization is, by definition, difficult to articulate. Social realism deals with this issue by theorizing about mechanisms that would account for practice phenomena. However, social realism may simply represent a dilution of other inquiry positions without offering enough compensating strengths. A new methodology, based on reflexivity, is suggested. The researcher is required to continually reflect on her/his relation to the research process, both in relation to the choice of method and in the gathering and interpretation of data. PMID- 8716802 TI - A 'domains of practice' approach to the standards of psychiatric and mental health nursing. AB - Standards have considerable influence on the practice of a discipline. As the written and explicit expectations of nurses held by their professional group, nursing practice standards serve as a basis for performance evaluation and as a clarification of the areas of nurses' accountability. Delineated here are the issues that the Canadian Federation of Mental Health Nurses faced as they attempted to identify and describe the nursing behaviours essential to this specialty area. A rationale for the 'domains of practice' approach is provided. The problems inherent in creating such a national document are addressed. Seven practice domains are described; the standards for two domains, the 'Helping' and the 'Effective Management of Rapidly Changing Situations', are outlined in detail. PMID- 8716803 TI - How can clinical supervision become a real vision for the future? AB - Clinical supervision is increasingly cited as being a necessary part of the work and development of all nurses. This paper discusses an educational programme offered to nurses in one particular Trust and outlines the course philosophy, content, organization and background to this development. Emphasis is placed on the importance of clinical supervision, the student-centred approach to course delivery, and the need to recognize the importance of adequate training and education of all nurses in this area. PMID- 8716804 TI - Literature and psychiatry: a mutually interested view. AB - This discussion paper addresses a perceived gap between nurses and literature. The author contends that nurses are suspicious of literature. As a consequence nurses are disadvantaged when works of fiction are dismissed in relation to their thinking and to elements of their professional practice in emotional work with the people who use mental health services. The author contends that a cloud of suspicion hangs over literature in many nurses' minds, and that a long conditioning of a practical, problem-solving discipline obviates the need to read widely. Therefore the practical necessity of distinguishing between good or bad books, well or badly-written books, and useful books is often not addressed. A parallel is to be drawn between a creative writer's construction of his/her characters and the psychiatric nurse/therapist's efforts to empathize with clients' experience, as either activity can be an emotional experience. PMID- 8716805 TI - The idea of partnership is fine, but what about perceptions? PMID- 8716806 TI - Nurses' and occupational therapists' perceptions of their roles. PMID- 8716807 TI - A national helpline needs your help. PMID- 8716808 TI - The management of wandering in older people with dementia. PMID- 8716809 TI - Clinical diaries: a compelling necessity. PMID- 8716810 TI - Special care unit: audit of time out, enforced segregation and seclusion, St Nicholas Hospital. PMID- 8716811 TI - The concept of empowerment. PMID- 8716812 TI - [Growing more human by caring. Self-acualization and acquisition of competences by adequately performing ones tasks]. PMID- 8716813 TI - [The art of communicating. Interdisciplinary communication]. PMID- 8716814 TI - [What is the Superior Course Specializing in Care of Adults and the ElderLy with Chronic Diseases]. PMID- 8716815 TI - [Osteoporosis, a hidden threat]. PMID- 8716816 TI - [The impact of euthanasia on families]. PMID- 8716817 TI - [Treatment of bladder cancer. New methods, new management]. PMID- 8716818 TI - [Palliative treatments in the service of life]. PMID- 8716819 TI - [Human life and United States' politics]. PMID- 8716820 TI - [The Saint John of God shelter for alcoholics]. PMID- 8716821 TI - Kansas court refuses to ban lay midwifery. PMID- 8716822 TI - OB clinic rejects patient refusing HIV test: nurse sued. Case in point: Atapka v. Perimeter OB-GYN Associates, Inc. 912 F. Supp. 1566--GA (1996). PMID- 8716823 TI - Legal case briefs for nurses. VA: claiming has: concurrent disability": court orders compp. payments continued; CA: nurses sue for wrongful termination: judgment against "unnamed defendant" void. PMID- 8716824 TI - Nurse orders bystander in E.R. to "stay put". Case in point: Jack v. State 671 So. 2d 26--LA (1996). PMID- 8716825 TI - Outcomes and incomes. PMID- 8716826 TI - What to do about Harry? AB - This case study about a patient labeled "noncompliant" illustrates a common scenario that home care nurses face in everyday practice. The frustration a nurse experiences when attempting to help a patient who rejects assistance is common and demoralizing and is discussed professionally in this study. A nurse attorney, a nurse manager, and a nurse ethicist respond to the author's case to give insight on the many ways in which this case can be viewed. PMID- 8716827 TI - Problematic standards: improving organizational performance through the assess and improve phases. PMID- 8716828 TI - The efficacy of Bobath neurodevelopmental interventions at home for patients after postcerebrovascular accidents. AB - The traditional approach to rehabilitating patients after cerebrovascular accidents encourages the patient to use only the unaffected side for all activities. This article presents the Bobath approach, which focuses on patients using both sides to regain former capabilities. The rationale and implementation of this method are outlined so that the home care nurse can learn how to use this method, used so commonly by rehabilitative therapists. PMID- 8716829 TI - Medication problems in the elderly: a home healthcare nurse's perspective. PMID- 8716830 TI - Beyond traditional nursing care: cultural awareness and successful home healthcare nursing. AB - Culture affects all areas of clients' lives, including their beliefs about health and illness. Home care nurses must consider cultural values in planning effective and culturally competent nursing care. Getting in touch with one's own heritage helps nurses to understand and appreciate the cultures of others. PMID- 8716831 TI - The emergency department and home care: strange bedfellows. PMID- 8716832 TI - Nurse-client alliance: a pattern of home health caring. AB - Forming nurse-client alliances is the process of partnering with clients to elicit desired outcomes. Developing an alliance between nurse and client calls for contracting toward mutually agreed upon goals, a practice that can be satisfying to both. This article explains the alliance concept and applies it to real-life situations so the nurse can integrate this strategy into practice. PMID- 8716833 TI - Evaluating the quality of home care services using patient outcome data. AB - Home healthcare nurses increasingly must be focused on the outcomes of patient care. As home care is called on to deliver more services in fewer visits, it is important that clinical staff understand the concepts and apply them to patient situations. This article helps the home care nurse to understand the frameworks being used throughout the country in evaluating patient outcomes and how one agency used this framework to evaluate care. PMID- 8716834 TI - The usefulness of the HCFA form 485 in predicting home healthcare costs. AB - As prospective pay systems are created, many sources of home care information are being considered. This study examines items on the HCFA Form 485 for their ability to accurately predict home healthcare costs. The researchers deduced that the current information available on the HCFA Form 485 was not useful in anticipating, planning for, and, ultimately, controlling costs. PMID- 8716835 TI - Patient care conferences with pizazz. PMID- 8716836 TI - Measurement of outcomes: what does it really mean? PMID- 8716837 TI - A spirit-focused conceptual model of nursing for the advanced practice nurse. AB - A spirit-focused conceptual model of nursing views the person as a spiritual being whose life and purpose arise from a personal relationship with God. The individual's definition of God determines how he or she responds to stimuli, sets goals, and establishes purpose in life. Consequently, this definition influences the professional nursing relationship, and the nurse's and patient's concepts of health and perceptions of the environment. The pediatric nurse practitioner and client are the primary examples of practice examined in this study. PMID- 8716838 TI - Pediatric nurses' knowledge and practice related to infant pain. AB - A convenience sample of 181 registered nurses completed questionnaires about their knowledge, attitudes, and clinical practice related to infant pain. The majority of respondents thought infants felt pain with the same intensity as adults and were comfortable administering narcotic and nonnarcotic pain medications. The most frequently identified infant pain cues were: crying, irritability, and inability to be consoled. There were associations between greater numbers of pain cues and higher pain severity ratings and between postlicensure education about pain and number of strategies proposed to relieve pain. There were no associations for noncritical versus critical care unit assignment or participation in postlicensure education about pain with number of assessment cues. There were no associations for pain severity or noncritical versus critical care unit assignment with pain management strategies. Until the research base about management of infant pain is expanded, nurses should continue to use and evaluate a variety of pain management interventions on a case-by-case basis. PMID- 8716839 TI - Methods of newborn infant temperature monitoring: a research review. AB - Although research related to infant temperature measurement has been reported in the nursing literature since 1969, issues regarding site of thermometer placement, resultant normative values, and length of thermometer placement time in full-term and preterm infants still need clarification. This article reviews research findings relevant to temperature monitoring in the newborn, examines the impact of these findings on clinical practice, and identifies areas where further research is needed. PMID- 8716840 TI - The many faces of child sexual abuse. PMID- 8716841 TI - Indicators of child sexual abuse: children at risk. PMID- 8716842 TI - Parents as protectors. PMID- 8716843 TI - How will nurses make the transition to the information era? PMID- 8716844 TI - Continuity of care: discharge planning and community nurses. PMID- 8716845 TI - The scandalous neglect of mental health care ethics. PMID- 8716846 TI - Reflections on doing and writing interpretive research. PMID- 8716847 TI - Praxis methodology: action research without a group. AB - Critical action research is a compelling method for nursing research because it offers a sensitive and qualitative way to study change processes in action. Action research is of limited use to nurse researchers because it relies on group participation whereas most nursing work which we may want to research involves one to one relationships with clients. This paper presents a modification of critical action research. It is termed 'Praxis Research' and it is suitable for research projects that aim to empower individuals, rather than groups. The steps in the praxis research process are described as an on-going spiral of practice, self-reflection, scholarly inquiry and theorising which leads to changed practice. PMID- 8716848 TI - Pain management for children and infants. AB - In this overview of paediatric pain management the physiological and psychological effects of untreated pain are described. Reasons for the undertreatment of pain are identified as incorrect assumptions about children and pain; personal attitudes of families, nurses and physicians; inadequate research and training; and assessment difficulties. The problems of paediatric pain assessment are considered and an assessment strategy known as QUESTT is presented. Pharmacological and nonpharmacological interventions are discussed and the conclusion reached that despite the difficulties, infant and child pain can be controlled by using adequate nursing knowledge and treatment. PMID- 8716849 TI - Gerontic challenge. PMID- 8716850 TI - 'Doing clinical': the lived experience of nursing students. AB - This investigation looked at the phenomenon of 'doing clinical' from the nursing students' perspective. A qualitative descriptive research based on the phenomenological approach was chosen as the method to reveal the meaning of 'doing clinical', through the analysis of nursing students' descriptions of their everyday lived experience of undertaking the clinical program. Nine, second year Bachelor of nursing students with at least 13 weeks of clinical experience were interviewed. These intensive interviews were tape recorded and transcribed verbatim. A thematic analysis of significant statements of the text was conducted resulting in sixteen theme clusters deriving from specific themes. These sixteen themes clusters and specific themes fell into 6 categories and there were: being in a world of conflicting and confusing values, deciding whether nursing is for me, becoming a professional nurse, discovering what nursing is, being an active learner, and having mixed feelings and thoughts on entry to the clinical program. The essential structure of the meaning of 'doing clinical' from the students' perspective is described and implications for nursing education, practice and research is discussed. PMID- 8716851 TI - Environmental ways forward in a postmodern (nursing) world. AB - In this paper I am interested in showing how nurses' concerns for environmental issues reflect larger shifts in modern/postmodern thinking. Changes through premodern, modern, and postmodern times are highlighted. Focus is especially upon the natural world and damage to it in the name of human progress--from which humans also ultimately suffer. Nurses seeking to find environmentally sound ways forward in their practice are experiencing and enacting a postmodern 'mood', and by speaking out about the futility of perpetuating modernist environmental concepts, they are engaging in postmodern dialogue. Nurses recognise as well however, that when they challenge entrenched and environmentally damaging practices in their workplaces, they open themselves to disturbing currents of change in which stability and meaning may readily be lost. PMID- 8716852 TI - Staff rights. PMID- 8716853 TI - Passive patient? PMID- 8716854 TI - Infections in PD. PMID- 8716855 TI - 1995-1996 national directory of certified nephrology nurses. PMID- 8716856 TI - ANNA position statements. American Nephrology Nurses' Association. PMID- 8716857 TI - The Pooh theory. PMID- 8716858 TI - RNs being replaced by LPNs. PMID- 8716859 TI - Charting new ground. Interview by Helen Griffiths. PMID- 8716860 TI - Regulating the health professions. PMID- 8716861 TI - Self-regulation: a nursing tradition. PMID- 8716862 TI - Registered Nurses Association of British Columbia: position statement. Breastfeeding: promotion, protection and support. PMID- 8716863 TI - The made in B.C. approach. PMID- 8716864 TI - [Education and recycling for change. Reflections on an experience]. PMID- 8716865 TI - [Social accomplishments. Health education in school]. PMID- 8716866 TI - [Muscles of the thorax and the frontal part of the arm]. PMID- 8716867 TI - [Topical anesthesia by cold. Effects of its use on venipuncture with large caliber needles]. PMID- 8716868 TI - [Nursing research and research units]. PMID- 8716869 TI - [The ultimate purpose of a hospital is to resolve the health problems of the patients. Interview by Maria Jesus Nadal]. PMID- 8716870 TI - [It is not only my problem, it can happen to anybody. Interview by Maria Jesus Nadal]. PMID- 8716871 TI - [Prone position--a practical experience]. PMID- 8716872 TI - [Pathology of the back]. PMID- 8716873 TI - [Heparinized venous approaches]. PMID- 8716874 TI - [Hysteroscopy]. PMID- 8716875 TI - [Video projections. Effectiveness in nursing procedure demonstrations]. PMID- 8716876 TI - Testifying strategies: telling the truth as effectively as possible. PMID- 8716877 TI - Don't submit to a limiting label. PMID- 8716878 TI - Strategic alliances: the wave of the future. PMID- 8716879 TI - Strategies in third-party insurance lawsuits. PMID- 8716880 TI - Domestic violence: why do they stay. PMID- 8716881 TI - Reconceptualizing compliance in home health care. AB - The term "compliance" and its traditional definitions frequently are criticized in the literature. Although a number of authors advocate a collaborative model of compliance, with compliance decisions and responsibilities for health outcomes shared by provider and patient, few describe this model in practice. This study investigated compliance communication in two home care agencies. Findings from interviews with 6 nurses and observation during home visits to 25 patients (N = 31) revealed a prosocial, collaborative model of compliance that coincides with the participative model of medical care (Smith, 1989) and a redefinition of compliance advanced by Kontz (1989). PMID- 8716882 TI - Unveiling the mystique of gender dysphoria syndrome. AB - There are common myths surrounding the condition of gender dysphoria and individuals seeking therapy. The diagnosis of gender dysphoria syndrome is applied to individuals who feel ill at ease with their biological body and either seek hormonal reassignment therapy or hormonal therapy and sex reassignment surgery. The author discusses standards of care, diagnostic criteria, physiological and psychological changes incurred through hormonal therapy are introduced as well as surgical techniques. By assisting individuals with gender dysphoria syndrome to enhance the integration of their new gender role into their self-identity, nurses are in an excellent position to facilitate improvements in their overall health status as well. PMID- 8716883 TI - Presence: four ways of being there. AB - Presence is an important but confusing concept in nursing. At times, it is used to simply characterize a nurse's physical presence, while, at other times, it is used in a highly metaphysical sense to depict a nurse's full physical, psychological, and spiritual presence. As a concept, presence has not been clearly defined. Its core characteristics need to be identified and separated from those characteristics that reflect its variability. The authors establish a definition of presence as "being there" based on the essence of the concept as it is used in the nursing literature; identifying four ways in which presence varies based on the quality of being there, the focus of the nurse's energy, and the nature of the nurse-patient interaction; and address possible outcomes of each type. PMID- 8716884 TI - Speaking up: a moral obligation. AB - As we rush around attending to the essentials of our lives (family, friends, clients, employers), what is left? Nursing Forum invites readers to engage in thoughts and activities that may awaken an untouched place. We hope these writings will kindle your personal involvement in something that was previously avoided--because of bias, fear, or uneasiness--in order to stretch your mind and spirit. The purpose of this paper is to explore the act of speaking up as a moral obligation and its relationship to moral courage and habit. The difficulties of speaking up and the consequences of silence are examined. The benefits of speaking up are raising self-respect, gaining courage, forming good habits and passing on that legacy. PMID- 8716885 TI - The new equation. PMID- 8716886 TI - Whistleblowing. PMID- 8716887 TI - Language links. PMID- 8716888 TI - A clarification on the properties of aprotinin. PMID- 8716889 TI - Yes, you can give an inservice--with ease. PMID- 8716890 TI - A care planning tool that proves what we do. PMID- 8716891 TI - Eating disorders--still a threat. PMID- 8716892 TI - Chemo quick guide: plant alkaloids. PMID- 8716893 TI - The absolute skinny on the new fat substitute. PMID- 8716894 TI - Getting your patient off a ventilator. PMID- 8716895 TI - Critical care drug update. Part II. PMID- 8716896 TI - Hypnotherapy, not just hocus-pocus. PMID- 8716897 TI - Switches to OTC require more of patients. PMID- 8716898 TI - Letting patients go. PMID- 8716899 TI - The dawning of the new millennium in 2001. PMID- 8716900 TI - Eco-wellness nursing: getting serious about innovation and change. AB - Eco-wellness nursing represents a creative, challenging and empowering approach for facilitating a sustainable future for people and their environments, focusing beyond positivistic explanations and solutions, towards a more emancipating eco centric praxis. Underlying this approach are the principles of 'wellness', 'holism', and a 'reflexive and sustainable eco-culture'. The goal is helping people understand how to sustain their wellness, including their environments. Negotiating or brokering change from a traditional nursing model will require a revision of focus in roles, attitudes, contexts and actions of nurses. Crucial in this transformation is greater nursing independence in the private marketplace, which may involve consultancies, collaborative ventures, utilization of new technologies such as multimedia communications, and targeted nursing oriented research and development. PMID- 8716901 TI - Nursing scholarship: sense and sensibility. AB - This paper explores the relationship between nursing theory, research and practice. It suggests that the frequently discussed difficulties in operationalizing, testing and implementing nursing theories are perhaps indicative of their lack of fit with clinical practice. The insistence of fitting theories within the four metaparadigm concepts-person, environment, health and nursing-forces nursing theory to an inappropriate level of abstraction and to a level that dilutes its relevance for clinical application. It is possible that, while these theories are useful as philosophies, they lack the clinical relevance that is a critical criterion for theory. Confounding the status quo, the present practice of teaching theory as fact within education and using theory to drive nursing curricula removes the tentative and guiding contribution that a theory contributes to a profession. A plea is made for research that supports the exploration and identification of nursing concepts and for the development of appropriate theory that will guide both nursing research and nursing practice. Since nursing is an applied discipline, this responsibility necessarily falls on the shoulders of researchers who, in partnership with clinicians, can explore, test and refine theory. PMID- 8716902 TI - The discursive construction of the role of the nurse in medication administration: an exploration of the literature. AB - This paper draws on postmodern thought to interrogate the literature relating to nurses and medications. An examination of the representation of a specific nursing activity in the literature-the administration of medications-reveals much about the way in which the role of the nurse is discursively constructed. This is particularly evident from an analysis of the procedures shaping that role. These procedures, which nurses themselves develop and institute as rules to guide nursing practice, can have the effect of reducing nursing work to a series of rituals which contribute to the discursive construction of the nurse's role in medication administration. PMID- 8716903 TI - Nursing research methodology: transcending Cartesianism. AB - Nurses involved in research are concerned with methodological issues. This paper explores the Cartesian debate that has polarized the discourse on nursing research methodology. It is argued that methodologies exclusively based on objectivism, one pole of the Cartesian debate, or subjectivism, the other, do not provide nurses with adequate research foundations to understand the complexity of the lifeworld of nursing practice. This paper provides nurse researchers with an alternative methodological perspective, Gadamerian hermeneutics, which is in harmony with the clinical world of nursing practice. PMID- 8716904 TI - Nursing in quality space: technologies governing experiences of care. AB - This paper challenges contemporary portrayals in the nursing literature of the spaces within which care of patients in hospital settings is conducted. Within the wider discourse of fiscal restraint on health care spending, professional nursing has cast its disciplined eyes on details of the nurse-patient relationship for the ostensible purpose of repairing that which is treated as individual failings of nurses to practice in ways prescribed by nursing theories. Set aside in this approach to the so-called 'problems' of nursing practice has been an examination of the conditions within which nurses come in contact with patients, and a critical recognition that such contacts represent skilled accomplishments of social action. In this paper, these conditions are treated as resources available to both nurses and patients to generate accounts for the nurse's presence as well as the nurse's absence. Examples from an ethnographic study of nursing practice are used to illustrate how the spaces operating between nurses and patients are not empty voids but are social spaces through which particular meanings about nursing care can be conveyed. PMID- 8716905 TI - Through the looking glass: reflections on the authorship and content of current Australian nursing journals. AB - Over the past three years, Australian nurses have witnessed a proliferation of locally published refereed nursing journals. This paper considers the refereed content published in four selected Australian nursing journals over a recent twelve month period. Material appearing in the Australian Journal of Advanced Nursing (AJAN), Contemporary Nurse, Nursing Inquiry and Collegian was categorized according to subject matter and authorship by gender and discipline. Findings indicate that articles pertaining to clinical practice, the practice of research, and professional nursing issues are well represented in the literature. These findings mark a change from earlier analyses of Australian nursing journals, which suggested that the epistemologies of other disciplines were over represented in Australian nursing journals, and thus exerted a powerful influence over the development of nursing theory and scholarship. The increase in the number of refereed Australian nursing journals is indicative of the dynamic state of Australian nursing, in terms of confidence, scholarship and clinical practice development. PMID- 8716906 TI - Rethinking cultural sensitivity. PMID- 8716908 TI - One nurse's journey from LPN to PHD. PMID- 8716907 TI - Coming to doctoral research: a journey from confusion to confidence. PMID- 8716909 TI - I have no tolerance for impaired nurses. Melba Lee-Hosey addresses drug abuse among nurses. PMID- 8716910 TI - Nighttime pain: a major problem for one out of three adults. Gallup survey says pain interrupts sleep. PMID- 8716911 TI - The thyroid and its disease. PMID- 8716912 TI - Need for education among patients with vaginal symptoms. PMID- 8716913 TI - A look at schizophrenia: what it is, what it does. PMID- 8716914 TI - I've looked at death and it isn't so bad. PMID- 8716915 TI - [To give life and rediscover hope]. PMID- 8716916 TI - [Taking care of patients with multiple traumas]. PMID- 8716917 TI - [Teaching and evaluation: a few myths to be verified]. PMID- 8716918 TI - [Placing an intravenous catheter for the better prevention of nosocomial infections]. PMID- 8716919 TI - [Reinforce the patient's responsibility]. PMID- 8716920 TI - [Power to the patients--and to ourselves]. PMID- 8716921 TI - [The exhaustion of mothers--report on the meeting of mother's consultants. The difficult parting with the mother's ideals]. PMID- 8716922 TI - [New trends in psychiatry and psychiatric nursing. Professional nursing care is in the wings]. PMID- 8716923 TI - [Giant steps in psychiatric nursing]. PMID- 8716924 TI - [The specifics of nursing]. PMID- 8716925 TI - [Investigation on the specific characteristics of nursing. Nurses in the mirror]. PMID- 8716926 TI - [Good bye to Michael]. PMID- 8716927 TI - Audit Commission unfair to fundholders. PMID- 8716928 TI - Where are the specialist cancer nurses? PMID- 8716929 TI - Nursing vital to cancer care. PMID- 8716930 TI - Chance for choice. PMID- 8716931 TI - A fresh approach to emergency care. PMID- 8716932 TI - Antithesis of care. PMID- 8716933 TI - Pre-operative patient instruction: is it effective? PMID- 8716934 TI - Nurses' concerns about i.v. therapy and devices. AB - This article reports the findings of a survey of nurses' knowledge and anxieties about intravenous (i.v.) therapy. The anonymised questionnaire survey canvassed nurses' views on education in the use of i.v. devices, complications arising from i.v. therapy, and which side-effects caused them greatest concern for patient welfare. The author reports widespread dissatisfaction with the level of education that is provided and cites published guidelines and reports which can help to inform nurses. PMID- 8716935 TI - Transactional analysis: how to improve communication skills. AB - Transactional analysis (TA) can be a useful framework for promoting effective communication. This article outlines the theoretical constructs of TA and suggests a strategy for teaching the model to diploma student nurses. The authors argue that an understanding of TA will allow nurses to analyse and adjust their skills to communicate more effectively. PMID- 8716936 TI - Nursing care of patients undergoing pallidotomy. AB - This article describes the pathology of Parkinson's disease, and how it can be managed through drug therapy and surgical intervention, particularly pallidotomy. A detailed account of the peri-operative nursing care of patients undergoing pallidotomy is given to highlight the demanding nature of the procedure for patients and staff. PMID- 8716937 TI - The management of eczema in adults (continuing education credit). PMID- 8716938 TI - Assessing information needs of patients' relatives: a study of Nehru Hospital. PMID- 8716939 TI - Teaching: a profile. PMID- 8716940 TI - John Barnbas Memorial Lecture. Social policy and social movement: the child in independent India. PMID- 8716941 TI - Acute care of the spinal cord injured. AB - In 1993, there were 583 new spinal cord injuries in Canada (Canadian Paraplegic Association, 1993). Seventy-five percent of those were males and sixty-seven percent were between the ages of 16 and 40. Motor vehicle accidents account for the highest percentage of injuries, while falls and sports are second and third respectively. The impact of this type of injury has many far reaching implications, not only on the individual and their family but also on society as a whole. These patients are surviving and living longer than before. Care of these patients has advanced considerably. Nurses caring for these patients must be experts in their field, having a solid understanding of the multiple system effects of traumatic spinal cord injury (SCI) and their implications. PMID- 8716942 TI - Pragmatics: right brain communication. AB - Neuroscience nurses as well as other health care providers are called upon to assess and implement intervention strategies to assist clients and family members and significant others to communicate in the presence of communication deficits. Language, high level language skills and speech are frequently addressed in presentations and in the literature but pragmatics which includes prosody, attitude, emotions and gestural behaviors is rarely addressed. Pragmatic competence mediated by the right hemisphere, is an equally important and essential component of human communication. An understanding of pragmatics permits the neuroscience nurse to identify and evaluate pragmatic deficits so interventions to facilitate communication can be undertaken. Specifically prosody, that is melody, pause, intonation, stresses and accents, affective tone, subtle grades of meaning and varied emphasis; its components; and aprosodia (lack of prosody) will be discussed. Kinesics, that is limb, body and facial movements, and referred to as pantomime and gestures, will be examined also as will facial expression. Suggestions for pragmatic assessment and therapeutic nursing interventions will be given. PMID- 8716944 TI - You make the diagnosis. School nursing: a child with asthma. PMID- 8716943 TI - Advanced concepts in the management of cerebral vasospasm associated with aneurysmal subarachnoid hemorrhage. AB - Despite advances in the diagnosis and surgical treatment of aneurysmal subarachnoid hemorrhage (SAH), the morbidity and mortality rates remain disturbingly high. In those patients that survive the initial event, deaths due to re-bleeding have been reduced as a result of a trend to earlier surgical repair of the aneurysm. Unfortunately, a large percentage of patients go on to die or become permanently disabled from cerebral infarction as a consequence of vasospasm. The exact mechanisms that contribute to the development of cerebral vasospasm in these patients continue to elude researchers. The pathological processes implicated in the vasospasm phenomenon will be described. Even though no single pharmacological agent or treatment protocol has been identified that prevents or reverses this deadly complication, a number of promising intervention strategies are being employed in the critical care setting. These will be explored in light of the most current research findings. Clearly, early detection and prompt treatment are vital if the outcomes for persons suffering aneurysmal SAH are to improve. A sound knowledge base, well-developed assessment and clinical reasoning skills, and a deeper understanding of the pathological processes and treatment objectives will assist the neuroscience practitioner to more ably care for these most challenging patients. PMID- 8716945 TI - "Bowling alone" in the age of electronic communication. PMID- 8716946 TI - Fatigue: a summary of the definitions, dimensions, and indicators. AB - Fatigue is a universal complaint that may or may not be related to medical diagnoses or therapeutic treatment. Fatigue is a complex, multicausal, multidimensional, nonspecific, and subjective phenomenon for which no one definition is widely accepted. The absence of theories concerning the nursing diagnosis of fatigue limits insight into the phenomenon and hinders the development of valid and reliable instruments for the measurement of fatigue. Therefores, the authors have undertaken a concept of analysis of fatigue, to a) document how it is defined in the literature; b) which dimensions of the concept of fatigue are distinguished; and c) which indicators of both the content and forms of fatigue represent the concept in practice. This concept analysis has resulted in a theoretical frame of reference that can indicate directions not only for further research into the nursing diagnosis of fatigue, but also for the development of measuring instruments. PMID- 8716947 TI - Validation of the defining characteristics of the nursing diagnosis ineffective airway clearance. AB - The investigators designed this validation study to determine the defining characteristics of the nursing diagnosis ineffective airway clearance. Using Fehring's Diagnostic Content Validity Model, 546 nurses who care for respiratory clients validated one major and 19 minor defining characteristics. Nine characteristics previously described in the literature were eliminated. Clarification of the defining characteristics of this diagnosis improves communication in practice, education and research. PMID- 8716948 TI - Diagnostic language consistency among multicultural English-speaking nurses. AB - Cultural differences among nurses may influence the choice of terminology applicable to use of a nursing diagnostic statement. This study explored whether defining characteristics are consistently applied by culturally varied nurses in an English language setting. Two diagnoses, pain, and high risk for altered skin integrity, were studied within six cultures: African, Asian, Filipino, East Indian, African-American, and Anglo-American nurses. Overall, there was consistency between the cultural groups. Analysis of variance for the pain scale demonstrated differences among cultures on two characteristics of pain, restlessness and grimace. The only difference on the high risk for altered skin integrity scale was found on the distructor, supple skin. PMID- 8716949 TI - A phenomenological description of the experience of spiritual distress. AB - The purpose of this study was to describe the phenomenon of spiritual distress in order to clarify the nursing diagnosis of spiritual distress and to evaluate its defining characteristics. Phenomenological interview procedures were used. Ten participants were asked to describe a time in their life when they had been concerned about the meaning of life, death, and/or their beliefs (the major defining characteristics of this nursing diagnosis). The data were analyzed using a hermeneutical approach. The thematic structure of this experience indicated a biphasic process through which participants moved from distress to change and growth. Implications for nursing are discussed and suggestions made for further research on spiritual distress. PMID- 8716951 TI - Nursing care of a patient with chronic abdominal pain secondary to porphyria: a case study. AB - Patients with porphyria present a wide variety of challenges to healthcare providers. The nursing care required by patients with porphyria is similar to that needed by patients suffering from chronic illnesses. This nursing care includes pain management, management of nutrition, an appropriate bowel regimen, the recognition of possible complex neurological manifestations, and psychosocial support (Shively et al., 1994). A thorough understanding of the disease process is essential to accurately assess the patient and develop an effective plan of care. PMID- 8716950 TI - Working together works. PMID- 8716952 TI - Pediatric constipation. AB - The purpose of this article is to present an overview of pediatric constipation. Discussion of the definition from a medical and nursing point of view is included. Intestinal pathophysiology as well as etiological theories of pediatric constipation are reviewed. Current research to date and clinical treatment and experience in the area are presented. A rationale for further nursing research in this area is described. PMID- 8716953 TI - Healthcare workers and bloodborne pathogens: knowledge, concerns, and practices. AB - Occupational exposure of healthcare workers to bloodborne pathogens and the transmission of human immunodeficiency virus and hepatitis B virus warrant study. This study examined issues related to the transmission of bloodborne pathogens in practice. Findings provide information regarding knowledge level, practices of staff in applying universal precautions, and availability of supplies and equipment. The concerns of nurses along with content for educational programs and suggested approaches to education are outlined. PMID- 8716954 TI - Gilbert syndrome. AB - Two or three times a year, a healthy patient (> 1 year old) with mild unconjugated hyperbilirubinemia as the only abnormality on routine liver function testing will be referred to the gastroenterologist. A diagnosis of Gilbert Syndrome (GS) is made, and the patient may or may not be seen in the clinic or office because GS is a benign condition. Either way, the nurse needs to understand the pathophysiology of this condition so he/she can provide necessary patient/family education. The purpose of this article is to review the existing literature on GS and to summarize it in a manner useful to both nurses and families. PMID- 8716956 TI - Reprocessing of flexible gastrointestinal endoscopes--an American Society for Gastrointestinal Endoscopy white paper. PMID- 8716955 TI - Performance of flexible sigmoidoscopy by a clinical nurse specialist. AB - According to the National Cancer Institute, 138,200 new cases and more than 55,000 deaths from colorectal cancer occurred in the U.S. in 1995. A high percentage of early cancers can be detected by screening asymptomatic persons older than age 50 with a digital rectal exam, fecal occult blood testing, and sigmoidoscopy. Evidence suggests that a decrease in mortality from colorectal cancer is associated with regular screening by sigmoidoscopy in an average risk population. As the population ages, more persons are eligible for colorectal cancer screening. Physician endoscopists are not available in all areas, and changes in reimbursement rates have caused some to discontinue this service. For these reasons, the Veterans Administration Medical Center in Buffalo, NY, developed a Colorectal Cancer Screening Program managed by a gastroenterology Clinical Nurse Specialist (CNS). Through study and supervision, the CNS has become proficient in the performance of 60-cm flexible sigmoidoscopic techniques and the diagnosis of multiple colon disorders, including sessile and pedunculated polyps, diverticulosis, tumors, hemorrhoids, and other conditions. PMID- 8716957 TI - Glucophage (metformin hydrochloride). PMID- 8716958 TI - [Protocols of agreement and teaching regulations]. PMID- 8716959 TI - [Protocols of agreement and teaching regulations: but do we have to do it all anew?]. PMID- 8716961 TI - [Health policies for a healthier society. From a trip for documentation study in Finland]. PMID- 8716962 TI - [Proposal to introduce a permanent education service into the public health service, local number 3, in Nuoro]. PMID- 8716960 TI - [Nursing sciences as an academic discipline at the Finland University, a base for the assurance of good quality in nursing]. PMID- 8716963 TI - [The concept of need in nursing care]. PMID- 8716964 TI - [Patient information at the moment of admission to the hospital. Its use as shown in an investigation using questionnaires]. PMID- 8716966 TI - Public outcry greets plans for 'generic worker'. PMID- 8716965 TI - [Virginia Henderson, "the most loved nurse in the world" died at the age of 98 years]. PMID- 8716967 TI - Pressure on sponsored nurses. PMID- 8716968 TI - Change at last? PMID- 8716969 TI - Rebuild trust. PMID- 8716970 TI - I can do that. PMID- 8716971 TI - Accept the new. PMID- 8716972 TI - When Ecstasy means agony. PMID- 8716973 TI - New success for old treatment. PMID- 8716974 TI - Encouraging excellence. PMID- 8716975 TI - Incidence of infection after caesarean section: a study. AB - This article reports a study of the incidence of infection in patients undergoing Caesarean section in a district general hospital maternity unit. A literature review was carried out and data were collected on operating circumstances, catheterisation, rates of infection, antibiotic therapy and post-discharge infection surveillance. A high incidence of infection was discovered and there was no evidence of a systematic approach to antibiotic therapy. The author highlights the need for further research on a larger group of patients and describes changes in practice which have occurred locally since completion of the study. PMID- 8716976 TI - Nursing dependency scoring: measuring the total workload. AB - This article discusses the issues surrounding the measurement of nursing work. The authors highlight a Royal College of Nursing (RCN) survey of critical care units which identified the extent of the use of patient scoring systems. The authors also examine the implications of using scoring systems to determine nursing workload and estimate staffing requirements. PMID- 8716977 TI - Clinical skills laboratories: teaching practical nursing. AB - This article describes the development and subsequent use of a clinical skills laboratory in one school of nursing and midwifery. It aims to allow students more time to practise psychomotor skills which clinicians, managers, educationalists and students themselves have perceived to be lacking in students since the implementation of Project 2000. The author suggests that this teaching strategy facilitates skill acquisition, increases students' confidence, and may in the long term help to bridge the theory-practice gap. PMID- 8716978 TI - Rheumatoid arthritis (continuing education credit). PMID- 8716979 TI - Interferon: an overview for dermatology nurses. AB - Interferons are glycoproteins that have numerous immunologic effects. Over the last decade the use of interferon has grown dramatically. Interferon preparations are now commonly used in many dermatologic conditions. The types of interferon that are commercially available, their therapeutic applications, and the nursing care of patients who receive interferon are reviewed. PMID- 8716980 TI - The value and limitations of protective gloves in medical health service: Part I. AB - The value of protective gloves in the health care environment is well known. However, these gloves are not perfect. Some are permeable to microorganisms and various chemicals, and/or result in side effects for health care professionals. Gloves that offer the greatest protection and least side effects are highly desirable. Field of application rules and regulations, materials and manufacturing, and test methods of protective gloves are described in Part I of this three part series. PMID- 8716981 TI - What's your assessment? Porphyria cutanea tarda. PMID- 8716982 TI - Moist wound healing. AB - The optimum wound environment to enhance wound healing is a balance of nutrition, hypoxia, and removal of debris in an occlusive moist environment. With increasing knowledge of the healing process and the variety of dressings available, the end result of any wound management will be an expedited wound healing with maximum patient comfort. PMID- 8716983 TI - Lasers in dermatology: a nursing perspective. AB - The rapid growth in laser technology has led to an increasing number of conditions which are treatable with improved outcomes. Understanding the principles of lasers and the conditions which can potentially be treated allows the nurse to participate in all aspects of a laser practice from patient education to peri-operative care. PMID- 8716985 TI - Smoking cessation clinical practice guideline: implications for nephrology nurses. PMID- 8716984 TI - Federal health initiatives: a status report. PMID- 8716986 TI - President's message. Together, let's get the job done! PMID- 8716987 TI - Integrating research and practice for optimal clinical outcomes: an interview with Tim Porter-O'Grady. Interview by Gail S Wick. AB - Dr. Porter-O'Grady has written over 125 articles and book chapters and has published eight books and is completing a ninth. He has consulted with over 500 institutions and has spoken in 1300 settings in the U.S., Canada, Europe, and Asia and logs about 350,000 miles a year. Dr. Porter-O'Grady is listed in six different categories of Who's Who in America, serves on 7 editorial boards, and is a member of the New York Academy of Sciences and a Fellow in the American Academy of Nursing. He has served on a number of community boards and has been an elected officer in a variety of health related agencies and organizations. He is currently a health systems expert for the National Health Policy Council and is a member of the Georgia Health Care Reform Project. In this interview, which took place in Atlanta by ANNA Past President Gail Wick, Dr. Porter-O'Grady shares his wealth of knowledge and experience by challenging us to move beyond the old thinking of caring for a specific patient population to managing lives on a continuum of care in an interdependent relationship with other providers, to refine the term "patient care," letting go of the medical sickness model and returning to our life-centered, health-oriented nursing roots and to broaden our educational preparation to a systems perspective and a continuum of caring. PMID- 8716988 TI - Diabetes insipidus. AB - Diabetes insipidus (DI), an acute or chronic condition, results from either of two situations: An inadequate secretion of antidiuretic hormone (ADH) from the posterior pituitary gland or an insufficient renal response to adequate levels of ADH. Characterized by massive urinary output despite progressive serum hyperosmolality, hypernatremia, and dehydration, DI can be life-threatening if not promptly diagnosed and appropriately managed. Individuals with renal disease, surgical patients, and persons with cerebral trauma are at risk to develop this condition. PMID- 8716989 TI - A holistic protocol for management of fluid volume excess in hemodialysis patients. AB - Patients on hemodialysis as treatment for end stage renal disease (ESRD), are at high risk for ineffective management of their therapeutic regimen because it entails pervasive changes in lifestyle. This article describes the experiences of five patients who participated in a nursing protocol of 8-10 therapeutic interviews associated with dialysis treatment over a 5-week period. The Dungan Model of Dynamic Integration provided the conceptual framework for holistic management strategies of teaching, counseling, and social support to assist patients to integrate required changes into their lifestyles. Fluid volume excess (FVE) was used as an indicator of ineffective management of therapeutic regimens. Presence of FVE, indicated by weight gain, presence of peripheral edema, and abnormal lung sounds were assessed before each dialysis treatment. Findings confirmed that these patients were at high risk for ineffective self-management and that the approach of augmented professional support used in the protocol was valuable to patients and families during this difficult transition. PMID- 8716990 TI - Sexuality and fertility concerns of dialysis patients. AB - OBJECTIVES: The purposes of this study were to identify the frequency of sexual and fertility dysfunctions and satisfactions experienced by patients on dialysis therapy and their relationships to life quality variables, and whether patients desired to or received instruction regarding possible sexuality and fertility problems. DESIGN: The study used a descriptive, survey design. SAMPLE/SETTING: A convenience sample was obtained from 7 hospital-based dialysis centers in the state of Iowa. METHOD: A mailed questionnaire, composed of 7 instruments, was used to survey demographic and medical variables, activity level, fertility and sexual dysfunction, intimacy patterns, sexual attitudes, life and health quality, and satisfaction with sexual relationships of patients treated with dialysis therapy. RESULT: Sixty-five percent of the subjects (n = 135) were dissatisfied with their sexual relationship, 40% no longer engaged in intercourse or sexual activity, yet only 25% had received instruction regarding possible dysfunctions. Subjects satisfied with their sexual relationships differed significantly (p < .05) from those who were dissatisfied in activity level, intimacy patterns, and life and health quality with frequency of arousal, orgasm, and acceptance of partner's advances significantly related to sexual satisfaction. CONCLUSION: The standards of care identified by the American Nephrology Nurses' Association regarding sexuality had not been achieved in the sample measured. PMID- 8716991 TI - Hematologic and erythropoietin responses to iron dextran in the hemodialysis environment. AB - OBJECTIVE: To investigate the hematologic and economic advantages of using iron dextran as the sole supplemental agent to safely increase and maintain hematocrit levels and iron availability while optimizing erythropoietin dosing in patients on chronic hemodialysis. DESIGN: Iron dextran 100 mg (2 ml) was administered i.v. slow push, undiluted three times per week, sometime during the last 30 minutes of each hemodialysis treatment, until a total required ml (determined by using the package insert's formula) was attained. Maintenance doses of either 25 or 50 mg per week (dependent upon body weight) were administered ongoing to compensate for dialytic and gastrointestinal blood losses. The analysis duration was 12 months. SAMPLE/SETTING: A prospective analysis was performed on 13 clinically stable hemodialysis outpatients in a rural community hospital-based dialysis facility (mean age 56.4 years ranging from 24-76; sample included 9 males, 4 females). METHODS: The means and medians were calculated for each variable: hematocrit, ferritin, transferrin saturation, and erythropoietin dose. A one-tailed paired student t test was performed on doses of erythropoietin at -1 and 6 months, -1 and 9 months, and -1 and 12 months. Cost per patient of iron dextran loading dose and maintenance, as well as cost savings from actual erythropoietin dose reductions, were calculated at 3, 9, and 12 months. Cost savings reflected the cost of iron dextran. RESULTS: After 6 months on the protocol, erythropoietin doses decreased an average of 3100 units per patient with an 8% increase in hematocrit and 66% and 78% increase in transferrin saturation and ferritin, respectively. Based on averages in actual reduced erythropoietin dosing, a savings of +5,070 per patient per year was realized. CONCLUSIONS: This analysis found the use of iron dextran in the hemodialysis setting to be an effective and economic means to maintain hematocrit values and iron availability while optimizing erythropoietin dosing. PMID- 8716992 TI - Epoetin alfa--focus on nursing case management. Case study of the anemic patient. AB - Case management is a multidisciplinary collaborative approach that emphasizes continuity of patient care. Nursing case management can lead to improvements in the quality of care while affording nurses the opportunity to increase their expertise, autonomy, and authority in the health care system. Treatment of the anemia of end-stage renal disease (ESRD) is used as an example to illustrate the implementation of these principles in the dialysis unit. PMID- 8716993 TI - Transient hepatitis B antigenemia in hemodialysis patients following hepatitis B vaccination. PMID- 8716994 TI - Infection control measures used to prevent spread of drug-resistant organisms in chronic hemodialysis units. PMID- 8716995 TI - Liver transplantation in alcoholics. PMID- 8716996 TI - Liver transplantation in alcohol-related end-stage liver disease. PMID- 8716997 TI - View on Australian clinical chairs. PMID- 8716998 TI - The politics of pap smears. PMID- 8716999 TI - Nursing as healing work. PMID- 8717000 TI - Nine steps to better nursing management of incontinence. PMID- 8717001 TI - Writing qualitative research for publication. PMID- 8717002 TI - Promoting purposeful partnerships. AB - The clinical experience component of university based nursing courses has often attracted criticism from students, faculty and clinical agency staff. One way of addressing these difficulties is for stakeholders to engage in open and consistent dialogue. To this end, faculty from the School of Nursing at Griffith University (Gold Coast) initiated a trial of reflection sessions attended by students, clinical facilitators, faculty and clinical agency staff at the completion of each clinical experience. While the outcomes were essentially positive, personal, professional and political risks were revealed as inherent in the reflective process. Awareness of such risks is essential for the development of purposeful partnerships in clinical education. PMID- 8717004 TI - A difficult death. PMID- 8717005 TI - Nurses and health promotion: we really can make a difference. PMID- 8717003 TI - Emotional labour of nurses. AB - In the development of nursing as a profession there is a need for nurses to discuss the strategies they use to meet the emotional demands of the nurse patient relationship. These demands have changed over time. Hochschild's definition of emotional labour is used as a means to understand the content of nurses' emotional work. It is important that positive and supportive ward atmospheres are experienced by beginning nurse practitioners while they learn to cope with the emotional work involved in nursing. Emotional labour continues to be a very challenging aspect of nursing practice. PMID- 8717006 TI - Storytelling: a teaching-learning technique. AB - Nurses' stories, arising from the practice world, reconstruct the essence of experience as lived and provide vehicles for learning about nursing. The learning process is forwarded by combining storytelling and reflection. Reflection represents an active, purposive, contemplative and deliberative approach to learning through which learners create meaning from the learning experience. The combination of storytelling and reflection allows the creation of links between the materials at hand and prior and future learning. As a teaching-learning technique storytelling engages learners; organizes information; allows exploration of shared lived experiences without the demands, responsibilities and consequences of practice; facilitates remembering; enhances discussion, problem posing and problem solving; and aids understanding of what it is to nurse and to be a nurse. PMID- 8717007 TI - Assessing needs of people with cancer. AB - The provision of complementary cancer care is a developing field. Recent surveys into the provision of complementary therapies have targeted not only the consumer but also the purchasers and providers of health care. There is interest amongst health authorities regarding the provision of complementary medicine as a cost effective intervention in the management of chronic conditions. There are a number of recent studies which aimed to address this issue. The Royal London Homoeopathic Hospital NHS Trust (RLHH), offers homoeopathy and other complementary therapies to support people living with cancer. A Macmillan Nurse, specialising in Complementary Therapies, was appointed at the RLHH to further develop the program of complementary cancer care. A patient survey performed to assist the development of the program showed that people living with cancer most wanted psychological help from complementary therapies. Since the provision of designated programs of complementary cancer care, expansion in the range of therapies offered has taken place. PMID- 8717008 TI - Culturally appropriate for all? PMID- 8717009 TI - computer networking. PMID- 8717010 TI - Non-conventional topical therapies for wound care. AB - There seems to be a resurgence of interest among healthcare professionals in treating wounds with various topical agents. There are practitioners who are recommending and/or using topical therapies for wounds without having an understanding of why or how they affect a wound. Many of these products are used based upon here-say or clinical observations, since there is a lack of researched based practice regarding the use of such topicals in wound healing. This article reviews such non-conventional topical therapies as aloe vera, antacids, benzoyl peroxide, collagen, dilantin, gentian violet, impregnated gauzes, insulin, mercurochrome, oxygen therapy, sugar, and vinegar. Each agent is described, its mode of action is discussed, and clinical studies regarding the use of these topicals to promote wound healing are examined, if any. The reader should review this article along with other existing literature regarding topical therapies before making a decision about using such in the care of wounds. When caring for wounds, one should ask, what is the goal of wound care at this particular time? And then ask, based upon available research, what, if any, of these agents can help achieve this goal? The following information should assist one in answering these questions. PMID- 8717011 TI - Nurse caring behaviors: comparing five tools to define perceptions. AB - We asked which of five tools that measure perceptions of caring attributes (Larson's Care Q, Larson's Care/Sat, Nyberg's CAS, Duffy's CAT, Wolf's CBI) have the greatest usefulness in studies with patients and nurses. A convenience sample of 26 nurse administrators completed all five tools. Data was collected on completion time, demographic characteristics, and how results compared with earlier research using the same instruments. Larson's Care Q had similar results as previously described in the literature but problems included a forced choice format, difficulty with instructions, and it took the longest time to complete. Larson's Care/Sat was difficult to analyze, contained negatively worded items, but took the least time to complete. Nyberg's CAS was designed for use with nurses and would be difficult to use with patients. Duffy's CAT had negatively worded items. Similar results were obtained with Wolf's CBI as previously described in the literature and it had consistent language, short completion time, easy to understand instructions, and easy to analyze results which could be used in a correlational design study. We conclude that Larson's Care Q (using a simpler method of administration) or Wolf's CBI could be valuable in determining perceptions of caring in patients and nurses. PMID- 8717012 TI - Nutritional factors affecting wound healing. AB - The consistent relationship between poor nutritional status and risk of complications forms the cornerstone of nutritional support. Yet there is controversy about the ability of nutritional support to reduce complications or improve wound healing. This controversy stems from a number of issues. Diagnosing poor nutrition is not always easy and straight forward. There is sometimes a question whether a patient is malnourished or simply in overall poor health. Studies examining the relationship between nutrition and patient outcome are typically based on animal rather than human models. Even in clinical settings, aspects of care such as enteral or parenteral nutrient delivery may decrease the benefit of nutritional support, making outcomes even harder to measure. The effect of specific nutrients have been examined, such as protein, amino acids, vitamins C and A, and zinc. However, there are still questions regarding how much individual supplementation of a nutrient will positively affect overall outcomes. Although the relationship between specific nutrients and wound healing is not clearly defined by current studies, each patient should be provided with a complete, balanced therapeutic diet. There is at least suggestive evidence that improvement in nutritional status can improve outcomes of wound healing. PMID- 8717013 TI - [Industrial standards also for nursing?]. PMID- 8717014 TI - [Continuous improvements through the use of a quality commissioners]. PMID- 8717015 TI - [The reorganization of the operating room management]. PMID- 8717016 TI - [In the hospital as a patient]. PMID- 8717017 TI - [Following orders or refusal to act]. PMID- 8717018 TI - [Psychiatric home care services]. PMID- 8717019 TI - [On the way towards the 21st century. The management of the German Nursing Federation presents its yearly report]. PMID- 8717020 TI - [Nursing in pediatric oncology as shown for acute lymphatic leukemia]. PMID- 8717021 TI - American Nurses Association hails introduction of Patient Safety Act. PMID- 8717022 TI - A Proposition 187 in Florida: hazardous to public health. PMID- 8717023 TI - Study results show mother-infant time together can be increased with a change in infant's first bath. PMID- 8717025 TI - Nurse attorney notes. Documentation--an Rx for success. PMID- 8717024 TI - Through the eyes of a student. PMID- 8717026 TI - A comparative study of empathy of registered nurses based on academic preparation. PMID- 8717027 TI - Keeping faith in gene manipulation. PMID- 8717028 TI - Xenotransplantation trials 'should proceed but need guidelines'. PMID- 8717029 TI - US plan to scrap DNA panel splits genetic researchers. PMID- 8717030 TI - High standards urged for French charities. PMID- 8717031 TI - Something up Dawson's sleeve? PMID- 8717032 TI - A revolutionary philosopher of science. PMID- 8717033 TI - Smoking...harmful to the brain. PMID- 8717034 TI - Physiology: The acid test for band 3. PMID- 8717035 TI - Diabetes: Breaking the curse of the AGEs. PMID- 8717036 TI - A new member of the frizzled family from Drosophila functions as a Wingless receptor. AB - Receptors for Wingless and other signalling molecules of the Wnt gene family have yet to be identified. We show here that cultured Drosophila cells transfected with a novel member of the frizzled gene family in Drosophila, Dfz2, respond to added Wingless protein by elevating the level of the Armadillo protein. Moreover, Wingless binds to Drosophila or human cells expressing Dfz2. These data demonstrate that Dfz2 functions as a Wingless receptor, and they imply, in general, that Frizzled proteins are receptors for the Wnt signalling molecules. PMID- 8717037 TI - Entropically driven binding in a self-assembling molecular capsule. AB - The encapsulation of molecules within larger molecular or supramolecular cages (see ref. 1 for examples) brings about many interesting phenomena, among them the stabilization of reactive intermediates, new forms of stereoisomerism, and templating of the cage itself. Here we describe a self-assembling hydrogen-bonded molecular capsule that encapsulates guest molecules in a reversible, apparently entropy-driven process, giving rise to unusual temperature dependence of the formation process. The positive entropy of formation--at first sight unexpected- seems to be the result of the fact that more than one solvent molecule is included in the 'bare' cage, making their replacement by a single large molecule (here adamantane or ferrocene) entropically favourable. PMID- 8717038 TI - Role of leptin in the neuroendocrine response to fasting. AB - A total deficiency in or resistance to the protein leptin causes severe obesity. As leptin levels rise with increasing adiposity in rodents and man, it is proposed to act as a negative feedback 'adipostatic signal' to brain centres controlling energy homeostasis, limiting obesity in times of nutritional abundance. Starvation is also a threat to homeostasis that triggers adaptive responses, but whether leptin plays a role in the physiology of starvation is unknown. Leptin concentration falls during starvation and totally leptin deficient ob/ob mice have neuroendocrine abnormalities similar to those of starvation, suggesting that this may be the case. Here we show that preventing the starvation-induced fall in leptin with exogenous leptin substantially blunts the changes in gonadal, adrenal and thyroid axes in male mice, and prevents the starvation-induced delay in ovulation in female mice. In contrast, leptin repletion during this period of starvation has little or no effect on body weight, blood glucose or ketones. We propose that regulation of the neuroendocrine system during starvation could be the main physiological role of leptin. PMID- 8717039 TI - Consolidation in human motor memory. AB - Learning a motor skill sets in motion neural processes that continue to evolve after practice has ended, a phenomenon known as consolidation. Here we present psychophysical evidence for this, and show that consolidation of a motor skill was disrupted when a second motor task was learned immediately after the first. There was no disruption if four hours elapsed between learning the two motor skills, with consolidation occurring gradually over this period. Previous studies in humans and other primates have found this time-dependent disruption of consolidation only in explicit memory tasks, which rely on brain structures in the medial temporal lobe. Our results indicate that motor memories, which do not depend on the medial temporal lobe, can be transformed by a similar process of consolidation. By extending the phenomenon of consolidation to motor memory, our results indicate that distinct neural systems share similar characteristics when encoding and storing new information. PMID- 8717040 TI - Effects of nicotine on the nucleus accumbens and similarity to those of addictive drugs. AB - The question of whether nicotine, the neuroactive compound of tobacco, is addictive has been open to considerable scientific and public discussion. Although it can serve as a positive reinforcer in several animal species, including man, nicotine is thought to be a weak reinforcer in comparison with addictive drugs such as cocaine and heroin, and has been argued to be habit forming but not addictive. Here we report that intravenous nicotine in the rat, at doses known to maintain self-administration, stimulates local energy metabolism, as measured by 2-deoxyglucose autoradiography, and dopamine transmission, as estimated by brain microdialysis, in the shell of the nucleus accumbens. These neurochemical and metabolic effects are qualitatively similar to those of other drugs, such as cocaine, amphetamine and morphine, which have strong addictive properties. Our results provide functional and neurochemical evidence that there are specific neurobiological commonalities between nicotine and addictive drugs. PMID- 8717041 TI - Excitatory synaptic inputs to spiny stellate cells in cat visual cortex. AB - In layer 4 of cat visual cortex, the monocular, concentric receptive fields of thalamic neurons, which relay retinal input to the cortex, are transformed into 'simple' cortical receptive fields that are binocular and selective for the precise orientation, direction of motion, and size of the visual stimulus. These properties are thought to arise from the pattern of connections from thalamic neurons, although anatomical studies show that most excitatory inputs to layer 4 simple cells are from recurrently connected circuits of cortical neurons. We examined single fibre inputs to spiny stellate neurons. We examined single fibre inputs to spiny stellate neurons in slices of cat visual cortex, and conclude that thalamocortical synapses are powerful and the responses they evoke are unusually invariant for central synapses. However, the responses to intracortical inputs, although less invariant, are strong enough to provide most of the excitation to simple cells in vivo. Our results suggest that the recurrent excitatory circuits of cortex may amplify the initial feedforward thalamic signal, subserving dynamic modifications of the functional properties of cortical neurons. PMID- 8717043 TI - ICAM-2 redistributed by ezrin as a target for killer cells. AB - Very little is known about the receptors and target molecules involved in natural killer (NK) cell activity. Here we present a model system in which interleukin-2 activated killing by NK cells depends on the intercellular adhesion molecule ICAM 2 and is regulated by the distribution of ICAM-2. The level of ICAM-2 expression in NK-sensitive and resistant cells is similar, but in sensitive cells ICAM-2 is concentrated into bud-like cellular projections known as uropods, whereas in resistant cells it is evenly distributed. The cytoskeletal-membrane linker protein ezrin is also localized in uropods. Transfection of human ezrin into NK resistant cells induces uropods formation, redistribution of ICAM-2 and ezrin, and sensitizes target cells to interleukin-2-activated killing. These results reveal a new mechanism of target-cell recognition: cytotoxic cells recognize adhesion molecules that are already present on normal cells, but in diseased cells are concentrated into a biologically active cell-surface region by cytoskeletal reorganization. The results also highlight the importance of cytoskeletal interactions in the regulation of ICAM-2-mediated adhesive phenomena. PMID- 8717042 TI - Identification of the homologous beige and Chediak-Higashi syndrome genes. AB - Vesicular transport to and from the lysosome and late endosome is defective in patients with Chediak-Higashi syndrome (CHS) and in mutant beige (bg) mice. CHS and bg cells have giant, perinuclear vesicles with characteristics of late endosomes and lysosomes that arise from dysregulated homotypic fusion. CHS and bg lysosomes also exhibit compartmental missorting of proteins, such as elastase, glucuronidase and cathepsin G. Lyst, a candidate gene for bg, was identified by direct complementary DNA selection from a yeast artificial chromosome (YAC) clone containing a 650-kilobase segment of the bg-critical region on mouse chromosome 13. Lyst is disrupted by a 5-kilobase deletion in bg mice, and Lyst messenger RNA is markedly reduced in bg homozygotes. The homologous human gene, LYST, is highly conserved with mouse Lyst, and contains a frame-shift mutation at nucleotides 117 118 of the coding domain in a CHS patient. Thus bg mice and human CHS patients have homologous disorders associated with Lyst mutations. Lyst encodes a protein with a carboxy-terminal prenylation motif and multiple potential phosphorylation sites. Lyst protein is predicted to form extended helical domains, and has a region of sequence similar to stathmin, a coiled-coil phosphoprotein thought to act as a relay integrating cellular signal response coupling. PMID- 8717044 TI - Phosphorylation-dependent activation of the Ras-GRF/CDC25Mm exchange factor by muscarinic receptors and G-protein beta gamma subunits. AB - Muscarinic receptors activate Ras through a pathway distinct from that mediated through translocation of the exchange factor mSos1 by receptor tyrosine kinases. Here we report that muscarinic receptors can activate another Ras exchange factor, CDC25Mm, or p140Ras-GRF (refs 5,6). In NIH-3T3 cells expressing subtype 1 human muscarinic receptors (hm1), the agonist carbachol selectively increased the specific activity and phosphorylation state of epitope-tagged Ras-GRF. This stimulation was reversed by protein phosphatase 1 (PP1), and prevented by transducin alpha-subunits. Carbachol treatment of neonatal rat brain explants increasd Ras exchange factor activity and the phosphorylation state of endogenous Ras-GRF. In COS-7 cells, cotransfection of hm1 or hm2 receptors with Ras-GRF conferred carbachol-dependent increases in exchange-factor activity, whereas cotransfection with G-protein beta gamma subunits caused a constitutive activation that was sensitive to PP1. These results demonstrate a G-protein coupled mechanism for Ras activation, mediated by p140 Ras-GRF. PMID- 8717045 TI - Role for c-Abl tyrosine kinase in growth arrest response to DNA damage. AB - The c-Abl protein tyrosine kinase is activated by certain DNA-damaging agents, and its overexpression causes arrest in the G1 phase of the cell cycle by a mechanism dependent on the tumour-suppressor protein p53 (refs 2-4). Here we investigate the possible role of c-Abl in growth arrest induced by DNA damage. Transient transfection experiments using wild-type or inactivated c-Abl show that both induce expression of p21, an effector of p53, but only wild-type c-Abl downregulates the activity of the cyclin-dependent kinase Cdk2 and causes growth arrest. Exposure to ionizing radiation of cells that stably express active or inactive c-Abl is associated with induction of c-Abl/p53 complexes and p21 expression. However, cells expressing the dominant-negative c-Abl mutant and cells lacking the c-abl gene are impaired in their ability to downregulate Cdk2 or undergo G1 arrest in response to ionizing radiation. We also show that expression of c-Abl kinase in p21(-1-), but not in p53(-1-), cells results in downregulation of Cdk2. Our results suggest that c-Abl kinase contributes to the regulation of growth arrest induced by ionizing radiation by a p53-dependent, p21 independent mechanism. PMID- 8717046 TI - An agent cleaving glucose-derived protein crosslinks in vitro and in vivo. AB - Glucose and other reducing sugars react with proteins by a nonenzymatic, post translational modification process called nonenzymatic glycosylation or glycation. The sugar-derived carbonyl group adds to a free amine, forming a reversible adduct which over time rearranges to produce a class of products termed advanced-glycation end-products (AGEs). These remain irreversibly bound to macromolecules and can covalently crosslink proximate amino groups. The formation of AGEs on long-lived connective tissue and matrix components accounts largely for the increase in collagen crosslinking that accompanies normal ageing and which occurs at an accelerated rate in diabetes. AGEs can activate cellular receptors and initiate a variety of pathophysiological responses. They modify an appreciable fraction of circulating low-density lipoproteins preventing uptake of these particles by their high-affinity tissue receptors. Advanced glycation has also been implicated in the pathology of Alzheimer's disease. Because AGEs may form by a pathway involving reactive alpha-dicarbonyl intermediates, we investigated a potential pharmacological strategy for selectively cleaving the resultant glucose-derived protein crosslinks. We now describe a prototypic AGE crosslink 'breaker', N-phenacylthiazolium bromide (PTB), which reacts with and cleaves covalent, AGE-derived protein crosslinks. The ability of PTB to break AGE crosslinks in vivo points to the importance of an alpha-dicarbonyl intermediate in the advanced glycation pathway and offers a potential therapeutic approach for the removal of established AGE crosslinks. PMID- 8717047 TI - Structure of Taq polymerase with DNA at the polymerase active site. AB - The DNA polymerase from Thermus aquaticus (Taq polymerase) is homologous to Escherichia coli DNA polymerase I (Pol I) and likewise has domains responsible for DNA polymerase and 5' nuclease activities. The structures to the polymerase domains of Taq polymerase and of the Klenow fragment (KF) of Pol I are almost identical, whereas the structure of a vestigial editing 3'-5' exonuclease domain of Taq polymerase that lies between the other two domains is dramatically altered, resulting in the absence of this activity in the thermostable enzyme. The structures have been solved for editing complexes between KF and single stranded DNA and for duplex DNA with a 3' overhanging single strand, but not for a complex containing duplex DNA at the polymerase active-site. Here we present the co-crystal structure of Taq polymerase with a blunt-ended duplex DNA bound to the polymerase active-site cleft; the DNA neither bends nor goes through the large polymerase cleft, and the structural form of the bound DNA is between the B and A forms. A wide minor groove allows access to protein side chains that hydrogen-bond to the N3 of purines and the O2 of pyrimidines at the blunt-end terminus. Part of the DNA bound to the polymerase site shares a common binding site with DNA bound to the exonuclease site, but they are translated relative to each other by several angstroms along their helix axes. PMID- 8717049 TI - Optical mapping: a novel, single-molecule approach to genomic analysis. PMID- 8717050 TI - The human obese (OB) gene: RNA expression pattern and mapping on the physical, cytogenetic, and genetic maps of chromosome 7. AB - The recently identified mouse obese (ob) gene apparently encodes a secreted protein that may function in the signaling pathway of adipose tissue. Mutations in the mouse ob gene are associated with the early development of gross obesity. A detailed knowledge concerning the RNA expression pattern and precise genomic location of the human homolog, the OB gene, would facilitate examination of the role of this gene in the inheritance of human obesity. Northern blot analysis revealed that OB RNA is present at a high level in adipose tissue but at much lower levels in placenta and heart. OB RNA is undetectable in a wide range of other tissues. Comparative mapping of mouse and human DNA indicated that the ob gene is located within a region of mouse chromosome 6 that is homologous to a portion of human chromosome 7q. We mapped the human OB gene on a yeast artificial chromosome (YAC) contig from chromosome 7q31.3 that contains 43 clones and 19 sequence-tagged sites (STSs). Among the 19 STSs are eight corresponding to microsatellite-type genetic markers, including seven (CA)n repeat-type Genethon markers. Because of their close physical proximity to the human OB gene, these eight genetic markers represent valuable tools for analyzing families with evidence of hereditary obesity and for investigating the possible association between OB mutations and human obesity. PMID- 8717051 TI - Mechanically stretched chromosomes as targets for high-resolution FISH mapping. AB - When used with metaphase chromosomes, fluorescence in situ hybridization (FISH) makes it possible to localize probes to individual chromosome bands and to establish the order of probes separated by > or = 2-3 Mb in dual-color hybridizations. We evaluated the use of mechanically stretched chromosomes as hybridization targets for increased mapping resolution. Mapping resolution was tested by pair-wise hybridizations with probes from the 1p32-p33 region, spanning distances from 20 to approximately 1500 kb. Probes separated by > or = 170 kb could be ordered relative to one another and to the centromere-telomere axis of the chromosome. The advantages of the technique are the simple procedure for preparing the slides, the straightforward interpretation of the results, and the ability to score the predominant order from < 10 stretched chromosomes. However, because of the variability of stretching from one sample to another, the calculation of actual physical distances between probes is not possible. To illustrate the utility of this method, we showed that the gene for receptor tyrosine kinase TIE lies centromeric to COL9A2, RLF, and L-MYC genes at 1p32. The use of mechanically stretched chromosomes provides < or = 10-fold increased mapping resolution as compared with conventional metaphase FISH. Thus, the technique effectively bridges the gap between metaphase mapping and ultra-high resolution mapping (1-300 kb) techniques, such as the DNA fiber FISH. PMID- 8717052 TI - A second locus for hereditary hemorrhagic telangiectasia maps to chromosome 12. AB - Hereditary hemorrhagic telangiectasia (HHT) or Osler-Rendu-Weber (ORW) disease is an autosomal dominant vascular dysplasia. Initial linkage studies identified an ORW gene localized to 9q33-q34 but with some families clearly excluding this region. A probable correlation in clinical phenotype between the 9q3-linked families and unlinked families was described with a significantly lower incidence of pulmonary arteriovenous malformations observed in the unlinked families. In this study we examined four unrelated ORW families for which linkage to chromosome 9q33-q34 has been previously excluded. Linkage was established for all four families to markers on chromosome 12, with a combined maximum lod score of 10.77 (theta = 0.04) with D12S339. Mapping of crossovers using haplotype analysis indicated that the candidate region lies in an 11-CM interval between D12S345 and D12S339, in the pericentromeric region of chromosome 12. A map location for a second ORW locus is thus established that exhibits a significantly reduced incidence of pulmonary involvement. PMID- 8717053 TI - Quantitative trait loci that modify the severity of spotting in piebald mice. AB - Mice homozygous for the recessive mutation piebald (s) exhibit a white-spotted coat caused by the defective development of neural crest-derived melanocytes. The severity of white spotting varies greatly, depending on the genetic background on which s is expressed. A backcross between two inbred strains of s/s mice that exhibit large differences in the degree of spotting was used to identify six genetic modifiers of piebald spotting on chromosomes 2, 5, 7, 8, 10, and 13. The loci differed in their spatial contribution to spotting on the dorsal versus ventral surfaces of mice; nonadditive interactions were observed between loci on chromosomes 2 and 5. This study underscores the power of using genetic analyses to identify and analyze loci involved in modifying the severity of phenotypic traits in mice. PMID- 8717054 TI - Genes and languages in Europe: an analysis of mitochondrial lineages. AB - When mitochondrial DNA sequence variation is analyzed from a sample of 637 individuals in 14 European populations, most populations show little differentiation with respect to each other. However, the Saami distinguish themselves by a comparatively large amount of sequence difference when compared with the other populations, by a different distribution of sequence diversity within the population, and by the occurrence of particular sequence motifs. Thus, the Saami seem to have a long history distinct from other European populations. Linguistic affiliations are not reflected in the patterns of relationships of mitochondrial lineages in European populations, whereas prior studies of nuclear gene frequencies have shown a correlation between genetic and linguistic evolution. It is argued that this apparent contradiction is attributable to the fact that genetic lineages and gene frequencies reflect different time perspectives on population history, the latter being more in concordance with linguistic evolution. PMID- 8717055 TI - Mouse galactokinase: isolation, characterization, and location on chromosome 11. AB - Elevated galactose levels can be caused by several enzyme defects, one of which is galactokinase. Galactokinase deficiency cause congenital cataracts during infancy and presenile cataracts in the adult population. We have isolated the mouse cDNA for galactokinase, which shares extensive amino acid sequence homology, 88% identity, with a recently cloned human galactokinase. It is expressed in all tissues examined. In an interspecific backcross analysis galactokinase maps to the distal region of mouse chromosome 11, a region that is homologous to human chromosome 17q22-25. The availability of the mouse gene provides an opportunity to make a knockout model for galactokinase deficiency. PMID- 8717056 TI - Regional assignment of 68 new human gene transcripts on chromosome 11. AB - We have tested 80 expressed sequence-tagged site (eSTS) markers assigned to human chromosome 11 by the Genexpress program on a panel of somatic cell hybrids containing parts of this chromosome, characterized by cytogenetic data, reference markers, and with respect to the Genethon microsatellite genetic map. Sixty-eight new gene transcripts have been assigned to 25 subregions, one of which was newly defined by five of the eSTS markers. The markers are distributed on the short and long arms in agreement with their physical length. The genic map thus obtained has been integrated with the cytogenetic, genetic, and disease maps. Two eSTS markers have been further mapped with respect to a yeast artificial chromosome (YAC) contig close to the brain-derived neurotrophic factor (BDNF) gene and thus provide potential candidate genes for the mental retardation phenotype of WAGR (Wilms' tumor, aniridia, genitourinary abnormalities and mental retardation) syndrome. Altogether, the 68 new gene transcripts localized here represent more than a threefold increase in the number of unknown regionalized genes that could reveal potential candidate genes for the numerous orphan pathologies associated with chromosome 11. PMID- 8717057 TI - 130 kb of DNA sequence reveals two new genes and a regional duplication distal to the human iduronate-2-sulfate sulfatase locus. AB - Deficiency of IDs activity results in Hunter Syndrome (mucopolysaccharidosis type II), a fatal X-linked recessive disorder. We report characterization of 28 cosmids around the IDS locus in Xq28. Four overlapping cosmids have been sequenced in their entirety generating a 130-kb contig. These studies show the fine structure of the IDS gene and identify an IDS pseudogene-like structure located 20 kb distal to the active gene. Two novel genes have also been identified in this sequence, and one of these genes is also locally duplicated. Both homologs are expressed, and a number of alternative transcript products have been characterized. The presence of a highly conserved pseudogene-like structure within a larger duplicated region close to the IDS gene has significant implications for the study of mutations at this locus. PMID- 8717058 TI - Generation of entire human papillomavirus genomes by long PCR: frequency of errors produced during amplification. AB - Recently, several improvements of traditional PCR techniques have facilitated the amplification of significantly longer DNA target sequences. Here we report an improved method for amplification of entire human papillomavirus (HPV) genomes. Using rTth DNA polymerase, XL (Perkin-Elmer, Foster City CA), and the accompanying XL PCR buffer system, we have successfully amplified 8-kb genomes from approximately 10 copies of input reference strain HPV16 DNA. This long PCR (LPCR) method was subsequently used to amplify the entire HPV16 genome from clinical specimens. The fidelity with which the rTth DNA polymerase XL amplified target sequences under our chosen amplification conditions was estimated by partial sequencing of cloned LPCR products generated from cloned reference strain HPV16 genomes. A region spanning the HPV16 E6, E7, and part of the E1 open reading frames (ORFs) was sequenced in 29 clones. A total of 33 nucleotide substitutions were observed in the 23.5 kb sequenced. This corresponds to an error frequency of approximately one error per 700 bases. Finally, LPCR methods were used to amplify entire, novel HPV genomes from clinical specimens. LPCR primer pairs were designed for amplification of seven potentially novel HPV types. Amplicons of approximately 8 kb were generated from five of the seven HPV types targeted. One of the LPCR-generated novel genomes, CP141, was subsequently cloned and a partial sequence was determined. PMID- 8717059 TI - Heteroplasmy in the control region of human mitochondrial DNA. AB - The sequencing of a 360-nucleotide segment of the mt DNA control region in a population survey revealed the presence of one individual having heteroplasmy in positions 16,293 (A and G) and 16,311 (C and T). This case indicates that heteroplasmy in humans may not be as rare as generally believed. PMID- 8717060 TI - A commentary on the practical applications of competitive PCR. PMID- 8717061 TI - Anteroventral wall of the third ventricle and dorsal lamina terminalis: headquarters for control of body fluid homeostasis? AB - 1. The subfornical organ, median preoptic nucleus and the organum vasculosum of the lamina terminalis (OVLT) are a series of structures situated in the anterior wall of the third ventricle and form the lamina terminalis. The OVLT and ventral part of the median preoptic nucleus are part of a region known as the anteroventral third ventricle region. 2. Data from many laboratories, using techniques ranging from lesions, electrophysiology, neuropharmacology, Fos expression, immunohistochemistry and receptor localization, indicate that the tissue in the lamina terminalis plays a major role in many aspects of body fluid and electrolyte balance. 3. The subfornical organ and OVLT lack the blood-brain barrier and detect alterations in plasma tonicity and the concentrations of circulating hormones such as angiotensin II and possibly atrial natriuretic peptide and relaxin. 4. This information is then integrated within the lamina terminalis (probably in the median preoptic nucleus) with neural signals from other brain regions. The neural output from the lamina terminalis is distributed to a number of effector sites including the paraventricular (both parvo- and magno-cellular parts) and supraoptic nuclei and influences vasopressin secretion, water drinking, salt intake, renin secretion, renal sodium excretion and cardiovascular regulation. PMID- 8717062 TI - AT1 receptor antagonist, TCV 116, does not prevent cardiac hypertrophy in salt loaded Dahl salt-sensitive rats. AB - 1. We investigated the effects of direct blockade of angiotensin II (AngII) by a potent, non-peptide angiotensin II toff 1 (AT1) receptor antagonist, TCV 116, on the development of cardiac hypertrophy in salt-loaded Dahl salt-sensitive rats. 2. Six week old male Dahl salt-sensitive rats (n = 44) were fed a 4% salt diet and were simultaneously given various doses of TCV 116 orally for 7 weeks. Each control group received vehicle alone. 3. Dietary high salt intake elevated blood pressure continuously and caused left ventricular hypertrophy in rats treated with vehicle. TCV 116 had a minor effect on the rise in blood pressure. Left ventricular mass (left ventricular weight/body-weight) decreased slightly but not significantly, in rats treated with 3 mg/kg per day of TCV 116 (n = 8). Higher doses of TCV 116 (6 and 10 mg/kg per day; n = 8 each) did not show a decrease in left ventricular mass compared with the vehicle control. 4. These results suggest that AngII blockade has only minor effects on hypertension and on cardiac hypertrophy in salt-loaded Dahl salt-sensitive rats and that the renin angiotensin system may not contribute to the development of cardiac hypertrophy in Dahl salt-sensitive rats on a moderately high-salt diet. PMID- 8717063 TI - Effects of hyperinsulinaemia on renal function and the pressor system in insulin resistant obese adolescents. AB - 1. In the present study, using the euglycaemic hyperinsulinaemic glucose clamp technique, we investigated the effects of hyperinsulinaemia on sodium-water metabolism and the pressor system in obesity, both of which have been reported to be closely associated with insulin resistance and/or hyperinsulinaemia. 2. Sixteen obese young subjects and 24 non-obese young subjects who were all normotensives, participated in this study. The 2 h euglycaemic hyperinsulinaemic glucose clamp was performed in a fasting state. The mean glucose infusion rate needed to maintain a fasting blood sugar level (FBS) during the last 30 min of the clamp was used as an indicator of insulin sensitivity (M-value). Before and after the clamp, the following parameters were measured: creatinine clearance (Ccr); urinary excretion of sodium (UNaV); fractional excretion of sodium (FENa); plasma renin activity (PRA); plasma aldosterone concentration (PAC) and plasma noradrenaline concentration (PNA). 3. The M-value was significantly lower in obese subjects compared with non-obese subjects, although FBS and fasting immunoreactive insulin levels were similar in both groups. UNaV and FENa fell only in obese subjects during the clamp, while Ccr showed no significant change in either group. PNA and PRA increased significantly and PAC tended to increase in both groups. 4. These results suggest that obese subjects have insulin resistance with respect to glucose metabolism, but that urinary sodium excretion and the pressor system remain insulin-sensitive; the sensitivity of the sodium retaining action to hyperinsulinaemia was actually higher in obese subjects than in non-obese subjects. Therefore, if compensatory endogenous hyperinsulinaemia was raised by insulin resistance, these two factors may lead to chronic sodium retention and pressor system stimulation and, in turn, to hypertension in obesity. PMID- 8717064 TI - Autoradiographic localization of active renin in the juxtaglomerular apparatus of the dog kidney: effects of sodium intake. AB - 1. The effects of dietary sodium intake on active renin binding in the juxtaglomerular apparatus (JGA) of superficial and juxtamedullary cortex of the dog kidney were examined by quantitative in vitro autoradiography using a radiolabelled renin inhibitor [125I]-H77, which has high affinity for dog renin. 2. Changes in sodium intake resulted in marked alterations of active renin binding in the radiolabelled JGA. In comparison with the control kidney (190.8 +/ 7.7 Bq/mm3), a higher density of binding occurred in the labelled JGA of sodium depleted kidney (277.7 +/- 6.2 Bq/mm3), while a lower density of binding was found in the labelled JGA of sodium-loaded kidney (99.3 +/- 7.4 Bq/mm3). 3. Active renin binding in the labelled JGA was significantly higher in superficial JGA than in their juxtamedullary counterparts, irrespective of sodium intake. 4. Pre-incubation with trypsin (0.5 mg/mL), a procedure known to activate prorenin, markedly increased active renin binding in the labelled JGA of control (+ approximately 35%; P < 0.01) and sodium-loaded kidneys (+ approximately 75%; P < 0.01), but had little effect on binding in the labelled JGA of the sodium depleted kidney (+/- approximately 5-10%; NS). The proportions of active renin as a percentage of total renin were 60, 75 and 95% in the labelled JGA of sodium loaded, control, and sodium-depleted kidneys, respectively. 5. Emulsion microscopic autoradiography revealed that the binding was exclusively localized in the JGA, including the afferent and efferent arterioles, macula densa and extraglomerular mesangium. Labelling extended to the interlobular arteries in sodium depleted kidney. 6. These results indicate that autoradiography combined with the in vitro binding of radiolabelled renin inhibitors may provide a useful tool to measure active and prorenin renin and thereby study the physiological regulation of renin in the kidney. PMID- 8717065 TI - Effect of steady shear stress on fluid filtration through the rabbit arterial wall in the presence of macromolecules. AB - 1. Transport properties of the arterial wall in general and endothelial cells in particular are known to be sensitive to fluid flow generated shear stress. 2. The effect of steady wall shear stress was studied in vitro on transmural water filtration in the presence of a control perfusate (10 mg/mL bovine serum albumin) and a test perfusate (50 mg/mL Dextran 500). The latter is a macromolecule with certain physical properties comparable to those of the circulating low density lipoproteins. Dextran 500 is a neutral polysaccharide carrying no charge groups. 3. Male rabbits were anaesthetized with sodium pentobarbital and one of the common carotid arteries was cannulated and excised. 4. Fluid flux across the artery wall was measured at a constant transmural pressure (150 cm H2O) in the presence and absence of luminal flow of the perfusate. 5. Luminal flow produced an average increase by 21% in the filtration flux relative to its value under static conditions with 10 mg/mL bovine serum albumin in the perfusate. In the presence of 50 mg/mL Dextran 500 in the perfusate an average reduction by 36% was observed in fluid flux when luminal flow was imposed in the artery. 6. The increase in flux with the control perfusate may be due to a structural alteration of the glycocalyx fibre matrix caused by flow imposed shear stress. The observed decrease in flux with 50 mg/mL Dextran 500 may be accounted for by the deformation of the dextran polymer chains when subjected to shear stress. Although intact molecules are likely to be excluded by the glycocalyx, parts of the chain may insert themselves in the network and enhance the steric exclusion of water. Also, decreased hydration of the medial interstitium may increase medial resistance to fluid transport. PMID- 8717066 TI - Distribution and anti-inflammatory effect of mesalazine on carrageenan-induced colitis in the rabbit. AB - 1. A controlled-release preparation of mesalazine microgranules (PentasaR; Ferring AS, Vanlose, Denmark) releases the active ingredient over a wide area from the small intestine to the rectum and is consequently expected to bring about therapeutic benefits to patients with ulcerative colitis and Crohn's disease. 2. Mesalazine microgranules (50 or 150 mg/kg per day) were administered orally to each rabbit with carrageenan-induced colitis for six weeks. Its inhibitory effect on colonic mucosal damage was assessed in terms of the microscopic damage scores, leukotriene B4 concentrations and concentrations of mesalazine derivatives. 3. At the end of the experiment, the mesalazine 150 mg group had gained a significantly greater bodyweight than the control group. Microscopic damage was significantly lower in the 150 mg group than in the untreated control group. Tissue concentrations of 5-aminosalicylic acid and acetyl-5-amino-salicylic acid in the small and large intestine were higher in the 150 mg group than in the 50 mg group. Mucosal leukotriene B4 levels tended to be lower in rabbits receiving the larger dose of mesalazine. 4. The present study indicates that slow release 5-amino-salicylic acid at the larger dose reaches the large bowel in sufficiently high concentrations following oral administration and significantly reduces carrageenan-induced colitis in the rabbit. PMID- 8717067 TI - Relative contributions of extracellular Ca2+ and Ca2+ stores to smooth muscle contraction in arteries and arterioles of rat, guinea-pig, dog and rabbit. AB - 1. These studies describe the functional effects of modulation of the sarcoplasmic reticulum (SR) Ca2+ stores at three levels of the vasculature: (i) large arteries (rat and guinea-pig aorta); (ii) small resistance arteries (rat tail artery, rabbit mesenteric artery, dog mesenteric artery); and (iii) arterioles (guinea-pig submucosal arterioles of the small intestine). 2. All tissues responded to phenylephrine (PE; 10 mumol/L) with a transient contraction in Ca(2+)-free Krebs', reflecting Ca2+ release from PE-sensitive Ca2+ stores. After pretreatment with cyclopiazonic acid (CPA; 30 mumol/L) or thapsigargin (TSG; 1 mumol/L), putative SR Ca2+ pump inhibitors, the PE-induced contraction in a Ca(2+)-free medium was significantly inhibited in arterial tissues at all levels of the vasculature. Similarly, ryanodine (RYA; 30 mumol/L), an agonist that enhances Ca2+ release from the SR, also reduced the PE contraction in a Ca(2+)-free solution. 3. CPA or TSG alone in the presence of extracellular Ca2+, caused marked and sustained contraction in the rat and guinea-pig aorta and marked but transient or no contraction in the resistance arteries. In the rat and guinea-pig aorta, RYA caused a slowly developing tension. Little increase in basal tension was produced by RYA in resistance arteries and arterioles. 4. The findings show that an agonist-releasable Ca2+ pool is present at all levels of the vasculature that is independent of the size of the vessels and suggest that under normal physiological conditions there is an intimate balance between the roles of the plasma membrane and of the SR in the maintenance of vascular contractility. It appears that the role of the SR diminishes as the arteries become smaller, while Ca2+ fluxes across the plasma membrane predominates. PMID- 8717068 TI - Ibudilast reduces intracellular calcium elevation induced by in vitro ischaemia in gerbil hippocampal slices. AB - 1. A microfluorometry was carried out to investigate the effect of 3-isobutyryl-2 isopropylpyrazolo[1,5-a]pyridine (ibudilast) on changes in levels of intracellular calcium concentration ([Ca2+]i) induced by in vitro ischaemia in the CA1 field of gerbil hippocampal slices. 2. When slices, loaded with a calcium ion sensitive dye (rhod-2) were exposed to a glucose-free physiological medium equilibrated with a 95% N2/5% CO2 gas mixture (standard in vitro ischaemia), a large [Ca2+]i elevation was detected approximately 5 min after the beginning of in vitro ischaemia. 3. When slices were perfused with the in vitro ischaemic medium containing 43 mumol/L ibudilast, a [Ca2+]i elevation was still observed; however, the extent of the increase in [Ca2+]i was significantly depressed in all subregions of the hippocampal slices. 4. The extent of this inhibitory effect of ibudilast on the in vitro ischaemia-induced [Ca2+]i elevation was in a similar range as those of Ca2+ blockers, including (+)-5-methyl-10,11-dihydro-5H dibenzo[a,d]cycloheptan-5,10-imine maleate (MK-801), flunarizine and dantrolene. 5. Similar [Ca2+]i increases in the CA1 field were induced by a Ca(2+)-free in vitro ischaemia, a high concentration of KCl or by specific agonists for glutamate receptor subtypes (N-methyl-D-aspartate (NMDA), (s)-alpha-amino-3 hydroxy-5-methyl-4-isoxazolepropionate (AMPA) and kainate); these increases were also depressed with 43 mumol/L ibudilast present in the perfusion medium. 6. These results indicate that ibudilast may act by depressing the Ca2+ accumulation during and shortly after ischaemia, a possible pharmacological action of ibudilast that leads to the amelioration of ischaemic injury in the central nervous system. PMID- 8717069 TI - Endothelin receptors in normal and diseased kidneys. AB - 1. Endothelin-1 (ET-1), the most potent vasoconstrictor yet identified, mediates a multitude of responses in various tissues including the kidney. The biological responses of ET-1 are mediated by specific cell surface receptors classified as ET(A) and ETB. Species differences are observed in the distribution as well as function of these ET receptors. 2. Involvement of ET has been demonstrated in a number of renal diseases, including ischaemia-induced acute renal failure, chronic renal failure, radiocontrast and cyclosporin-induced nephrotoxicity. ET antibodies as well as ET receptor antagonists have been shown to be beneficial in these disease models. PMID- 8717071 TI - Endothelins: renal tubule synthesis and actions. AB - 1. Renal tubules and, in particular, the inner medullary collecting duct, produce endothelin and express cognate receptors. 2. Endothelins inhibit vasopressin stimulated cAMP accumulation and water reabsorption in the collecting duct; endothelins may also inhibit sodium reabsorption in the proximal tubule and collecting duct. 3. Autocrine inhibition of sodium and water reabsorption in the inner medullary collecting duct by endothelin may play a role in maintaining extracellular fluid volume homeostasis. 4. Derangements in autocrine inhibition of sodium and water reabsorption in the inner medullary collecting duct by endothelin may be involved in the pathogenesis of the hypertensive state. 5. Nephron-derived endothelins may function in a paracrine manner to regulate interstitial, juxtaglomerular and vascular smooth muscle cell function. PMID- 8717070 TI - Signalling pathways activated by endothelin stimulation of renal cells. AB - 1. Endothelin mediates its effects in a variety of renal cells via a multiplicity of intracellular signalling pathways. 2. Stimulation of phosphatidylinositol specific phospholipase C (PI-PLC), resulting in the activation of inositol trisphosphate and diacylglycerol, can be detected even at picomolar concentrations of peptide. 3. Endothelin activation of cPLA2 is sensitive to ambient [Ca2+]i, is not contingent upon protein kinase C activation and is independent of PI-PLC stimulation, being coupled to the endothelin receptor in a yet to be determined manner. 4. Activation by endothelin of phosphatidylcholine specific phospholipase D is under the dual regulation of protein kinase C and [Ca2+]i, with protein kinase C being the major regulator and [Ca2+]i playing a secondary, modulatory role. 5. Phosphatidylcholine-specific phospholipase C (PC PLC) is stimulated by endothelin and accounts for the prolonged activation of diacylglycerol by this peptide. PC-PLC activity is critically dependent upon [Ca2+]i, whereas protein kinase C plays no role in modulating the activity of this enzyme. 6. Endothelin enhances the phosphorylation of protein tyrosine kinases, with evidence that phosphorylation of pp60 Src may be an important early event. PMID- 8717072 TI - Role of endothelin in renal function and dysfunction. AB - 1. Endothelin has significant effects on renovascular, glomerular and tubular function. 2. Endothelin causes severe renal vasoconstriction, resulting in a decrease in renal blood flow and glomerular filtration rate. 3. Endothelin can inhibit sodium reabsorption and, in the rat, vasopressin-induced water transport. 4. The endothelin receptor subtypes mediating renovascular and tubular effects of endothelin may differ between species. 5. Renal endothelin production, metabolism and receptor binding is altered in a number of renal diseases, including acute and chronic renal failure and cyclosporine and radiocontrast nephrotoxicity. 6. Endothelin receptor antagonists or antibodies can attenuate the severity or progression of a number of renal diseases. PMID- 8717073 TI - Endothelin is a key modulator of progressive renal injury: experimental data and novel therapeutic strategies. AB - 1. Glomerulosclerosis and tubulointerstitial damage are common histological abnormalities of many renal diseases that progress to end-stage renal failure. 2. In some models of renal damage, glomerulosclerosis seems to be associated with increased glomerular capillary pressure. 3. Due to the positive correlation of glomerulosclerosis and proteinuria in both experimental models and in humans, abnormal permeability to macromolecules has also been considered a possible determinant of glomerulosclerosis. 4. Abnormally filtered macromolecules have an intrinsic toxicity to the kidney due to protein over-reabsorption, possibly leading to tubulointerstitial damage. 5. Endothelin-1 (ET-1) is a vasoconstrictor peptide that induces mitogenesis and the accumulation of matrix proteins by mesangial cells. 6. Evidence is available that ET-1 plays a role in progressive renal disease in different experimental models, including renal mass reduction, lupus nephritis, streptozotocin-induced diabetes and puromycin-induced nephrosis. PMID- 8717074 TI - [Residency, between wish and reality]. PMID- 8717075 TI - [The ocular manifestations in systemic lupus erythematosus]. AB - Retinal involvement in systemic lupus erythematosus is dominated by a microangiopathy of unknown aetiology, with occlusive consequences. Although cases of severe retinal ischaemia have been recently described, from a statistic point of view, the occlusive phenomena seem to have declined over the last decades, under the influence of a better therapy and follow-up of the disease. The relatively high incidence of papilloedema and the fact that this oedema might be unilateral or asymptomatic are pointed out. The importance of ophthalmic examination in patients with SLE-like syndromes or in seronegative cases of SLE is revealed. Other ocular manifestation of the disease are reviewed. PMID- 8717076 TI - [The clinical and anatomicopathological aspects in malignant choroid melanoma]. AB - Choroidal malignant melanoma is the most frequently among the intraocular malignant tumors, and its incidence is getting hier. Clinical aspects (stadialisation, clinical forms) are strictly corelated with the anatompathological aspects (macroscopic exam, clasic histology, optic microscopy, electronic microscopy, TEM or scanning-SEM). Prognose and survival of malignant melanoma, depends on the histological form and on others microscopic changes. The results are integrated in a schema of conservative, complex, codifical, sequental treatement. PMID- 8717077 TI - [Prismatic and surgical block in nystagmus]. AB - After the speech of some teoretical notions (classification, ethyology, clinical description, treatment) about nystagmus are presented the summaries of dossiers of five patients consultated in our clinic with the diagnostic of nystagmus. At two from them realised the complete blockade at the contractions with prisms and at two from en the partial blockade too with prisms. One of them presenting nystagmus associate with strabismus was surgical solved. PMID- 8717078 TI - [The loss of a right muscle in the orbit intra- or postoperatively]. AB - One child four and half years old remains with a divergent deviation by forty five degree with absence of adduction, after an operation of convergent strabismus at the left eye. Considering that is about by one postoperatorial paralytic strabismus, computer tomography praises the right intern muscle retracted at the back of orbit, but it doesn't adheres at the eyeball. It considers that the divergent strabismus was caused by the slide of postoperatorial muscle. Looking middle line of the muscle, parallel with the middle wall of the orbit, it praises the right intern muscle retracted and after the excision of the granulation tissue all round, displays the muscle with its normal lenght. It cuts 3.5 milimeters from this muscle and it practices the retroposition of the extern right muscle with 7 milimeters. It obtains postoperatorial orthophoria. It talks about possibility of lossen of a right muscle in orbit intra- and post-operatorial and the measures that are imposed in these situations. PMID- 8717079 TI - [Ocular refraction in Marfan's syndrome and its surgical problems]. AB - The superior-temporal subluxation of the crystalline lens at the patients with Marfan syndrome makes optic problems who need a surgical solution. It describes one aspiration technique with concomitant support of crystalline lens, underlining the specifics features what are meet here: exessive hardness of crystalloid and the content of crystalline's sack. At eight operated eyes, the postoperatorial visual acuity restored at values over 5/10. PMID- 8717080 TI - [Ocular manifestations in Peters' syndrome]. AB - The analysis of twenty-one observations with Peter Syndrome, shows that the disease was established by a disorder in the development of the mesoderm who gives birth at the posterior, stratum of cornea, iris stroma and angle of the posterior chamber. The disease was associated with cataract (nine cases), strabismus (seven cases), congenital glaucoma (five cases), microophthalmia (three cases), microcornea (four cases), remainders of the pupillary membrane (one case), vitreous opacities (one case), facial malformations (one case), congenital dacryocystitis (one case), nystagmus (four cases). The presence at the same patient of the posterior keratoconus or staphiloma of cornea at one eye and Peter syndromme at the other eye, is possible to say that these diseases are evolutionary stages by one and the same mesodermal disgenesia. PMID- 8717082 TI - [Uveal inflammation secondary to lens implantation]. AB - On the basis of the 460 cases of implant of lens, the author analyses the different types of uveal inflammatory reactions appeared during the post operation period. These inflammations were more numerous and graver in the cases of implants with pupilar suport and of C.A. The implants of C.P. produced moderate inflammations and a reduced number of cases. The attentively supervising and the treatment carried out with steroidian anti-inflammatories (dexametazone, betametazone) or nesteroidians (diclofenac, indometacine) are able to stop the inflammatory reactions and to improve the functional results. PMID- 8717081 TI - [Genetic aspects in pigmentary retinopathy]. AB - The analysis of 123 cases with pigmentary retinopathy shows in 26 cases hereditary transmission with the existence of several affected members within the same family (21.2%). The isolated cases represented 78.8% of the total number. From the 26 familial cases 24 (19.5% from the total) presented a recessive autosomal transmission, 1 case presented a dominant autosomal transmission (3.84%), and 1 case presented a recessive sex-related transmission (3.84%). The research of the family tree is necessary in all the cases with pigmentary retinopathy, in order to find the heterozigotes which carry the pathological gene and to efficiently make the profilaxy of the disease. PMID- 8717083 TI - [Tuberculosis of the tarsal conjunctiva]. AB - A woman sixty-five old presents one pronounced swelling of the eyelids at the both eyes with the appearance of the multiple chalasions. The patient at twenty six years old presented pulmonary tuberculosis, in the present stabilized. The morphopathology exam by a fragment of tarsus and conjunctiva shows an lymphocyt inflammatory infiltration with the presence of the epithelial cells and caseation areas. At level of conjunctiva it constats the presence of tuberculum follicles that are present too at the level of Meibomius canals of excretions. The disease is considered like an allergical reaction at endotoxin of Koch bacillus, secondary by some old tubercular lesions. PMID- 8717084 TI - [The incidence of refractive errors in strabismus with an early onset]. AB - The paper represents one study effected at a number of two hundred and fifty children under three years old, examinated in Ophthalmological Clinic between 1992 and 1995. It was examinated strabismic deviation, visual acuity, state of binocular-eyesight refraction. Most of children (59%) present to hypermetropia under three dioptry; 23.3% medium hypermetropia (3-6 dioptries). Little percentage under 10% present others refraction vices: big hypermetropia, astigmatism, anisometropia, myopia. After optic correction and occlusion, most of deviations remained unaltered requiring surgical subsequent correction. This fact certifys the preponderence of neurogenic pathogenesis and not adjustment in strabismus with precocious debut. PMID- 8717085 TI - [Capsulotomy through the pars plana in the eye with pseudophakia of the posterior chamber]. AB - It described one surgical technique for capsulotomy in the case of secondary cataract at an eye with pseudophakic by posterior chamber. The approaching makes with an simple instrument, through pars plana, the tearing of posterior capsule making under control with operator microscope, in area of visual axie, excluding any possibility to harm the lens, its luxation in vitreous or anothers complications. PMID- 8717086 TI - [A case of corneal mycosis]. AB - A case of corneal ulcer in the right eye and iridocyclitis of the left eye, with mycotic ethiology in a 50 year woman who works in humid conditions with undefinite organic compounds. The diagnostic has been put through laboratory tests (direct frotee and culture). After the treatment with antimycotics the lesions have been epithelised, the patient leaving the hospital with good vision (VOD = 2/3, VOS = 1/3). After two years, in the picture of a sicca syndromme, the condition reappears in the left eye, the ulcer has perforated needing conjunctival covering. PMID- 8717087 TI - [Raverdino's operation, a procedure to resolve lacrimal dacryostenosis with deficient pituitary mucosa]. AB - The paper presents the efficiency of the Raverdino operatorial technique, which creates a large anastomosis between the lacrimal sac and nose, followed by an effective drainage in those cases with persistent obstruction of the nasolacrimal duct. The paper also exposes clinical cases in which the mentioned technique was used as first intervention or as reintervention after an inefficient dacryocystorhinostomy. The Raverdino technique offers a successful, long lasting result, without major intra- or postoperative complications. PMID- 8717088 TI - [Complications of retinal detachment surgery]. AB - The surgical treatment of the retinal detachment is concerning surgical technics, procedures, devices--both "classics" or from a new modern group, wich may lead sometimes to adverse reactions or complications. Not being always iatrogenic, their risck must be known under all their aspects, in order to addopt a proper and individualized therapy. PMID- 8717089 TI - [Complications specific to the types of pseudophakia]. AB - The paper presents some complications specific to the IOL implant corelated with the 3 main groups of pseudofac: anterior chamber, irian fixation and posterior chamber pseudofac. The introducing, the fixation type and the position occupied by every IOL type determines the more frequent occurrence of some complications specific to the pseudofac implant. The therapy of these complications varies too with the implant type. PMID- 8717090 TI - On hen egg fractionation: applications of liquid chromatography to the isolation and the purification of hen egg white and egg yolk proteins. AB - Liquid chromatography has been used as a means of egg protein analysis or as a method for the purification of egg proteins. Several chromatographic methods, including gel permeation, ion-exchange, reversed-phase, hydrophobic interaction, and immobilized-ligand-affinity chromatography, have been carried out for the separation or the purification of egg yolk or egg white proteins. Ion-exchange chromatography appears to be the most frequently used method for protein isolation and it is the easiest to adapt to a process scale. From an analytical point of view reversed-phase chromatography is, at the moment, the recommended method for egg white analysis. Egg white has been fractionated more often by liquid chromatography than has egg yolk. Several chromatographic methods have been developed on a laboratory scale, but the application of these techniques on an industrial scale remains limited. PMID- 8717091 TI - Effects of continuous flow microwave treatment on chemical and microbiological characteristics of milk. AB - Raw cows' and goats' milks were heated by microwave in a continuous flow unit up to temperatures ranging from 73.1 to 96.7 degrees C. The effects of the heat treatments were estimated by measurements of lactose isomerization, protein denaturation, inactivation of alkaline phosphatase and peroxidase and the total bacterial count. Negative phosphatase tests and low bacterial counts, together with low degrees of whey protein denaturation, were achieved under several temperature/time combinations. The results indicate that continuous microwave processing may be an efficient and mild approach for the pasteurization of milk. PMID- 8717092 TI - Organochlorine pesticide analysis in oil seeds and vegetable oil: simplification of silica gel clean-up and elimination of chlorinated solvents. AB - For the determination of organochlorine pesticides (OCPs) in vegetable oils and oil seed samples, a solid-phase extraction method is described, modified from that developed by Steinwandter, with silica gel columns activated with a 10% water content. In comparison to the method developed by Steinwandter, we reduced the amount of solvents and chemicals required for sample preparation and substituted dichloromethane by the much less toxic tert-butylmethyl ether in the eluent mixture. In this manner the sample preparation technique becomes more convenient, cheaper and ecologically more justifiable. Under the specified sample preparation and elution conditions 1 g silica gel retained about 16 mg triglycerides whereas the OCPs were quantitatively eluted. The recoveries for hexachlorobenze, (HCB) alpha-HCH, Lindane (gamma-HCH) gamma-HCH, Heptachlor, Heptachlorepoxid, o,p'-DDE, p,p'-DDE, Dieldrin, o,p'-DDT, p,p'-DDT were between 75 and 90%. The reproducibility of the total method was excellent as well as its ruggedness. PMID- 8717093 TI - Evolution and kinetics of monosaccharides, disaccharides and alpha-galactosides during germination of lentils. AB - The effect of light and seed rinsing during the germination of lentil seeds (Lens culinaris var. vulgaris, cultivar Magda-20) on the level of monosaccharides, disaccharides and alpha-galactosides (raffinose, ciceritol and stachyose) was investigated. The total soluble sugar content corresponded to about 9% of the mature seed weight, about 65% of which was alpha-galactosides. Germination brought about a large decrease in alpha-galactosides: 18% to 40% losses after 3 days and 100% after 6 days. However, glucose, which was not detected in ungerminated seeds, as well as fructose and sucrose gradually increased during germination. The content of alpha-galactosides decreased more rapidly when germinating seeds were given 6 h light per day, but under these conditions there was also a major reduction in the levels of fructose, glucose and sucrose. Seeds rinsed daily showed a greater reduction of alpha-galactosides and an increase in the levels of fructose, glucose and sucrose. When seeds were germinated for 10 days in the dark with daily rinsing, the content of alpha-galactosides decreased gradually during the first 4 days and they were not detected after 6 days. Under these conditions, fructose, glucose and sucrose, which represented about 3% of the mature seed weight, started increasing after day 2 and represented more than 13% of the germinated seed dry weight after 10 days. PMID- 8717094 TI - Thermospray-LC-MS analysis of various groups of polyphenols in tea. I. Catechins, flavonol O-glycosides and flavone C-glycosides. AB - This paper describes the application of thermospray-HPLC-MS (buffer ionization mode, single-stage MS, positive ion detection) to the analysis of flavanols (catechins), flavonol O-glycosides, flavone C-glycosides as well as caffeine, theobromine, theogallin and theanine from tea. All compounds are detected as pseudo-molecular ions [M+H]+. Other molecular ion species are adducts with sodium, potassium, ammonium and solvent clusters. The catechin gallates and the flavonol glycosides are fragmented. The fragmentation is temperature dependent. The glycoside bond is labile and consequently the flavonol glycosides have the protonated aglycone as this base peak. The ester bonds in the catechin gallates are more stable and the fragmentation is limited. The fragment pattern contributes to the structural information. LC-thermospray-MS is a good analytical tool for identifying the polyphenols mentioned above both in tea and other foodstuffs, especially in method development and structural elucidation. PMID- 8717095 TI - Combined effect of gamma-irradiation and conventional cooking on Aeromonas hydrophila in meatball. AB - Irradiation combined with a conventional cooking procedure was applied to meatball and the effects on bacterial load and inoculated Aeromonas hydrophila were determined. Meatball samples were irradiated by using a 60Co source at the dose levels of 0, 0.30, 0.75, 1.50, 2.50 kGy and cold stored at 4 +/- 1 degrees C for 7 days. Bacterial load and the count of A. hydrophila decreased when the irradiation dose level increased. A minimum inhibition effect was found at the dose of 0.30 kGy. Irradiation in combination with a conventional cooking procedure was found to be more effective in reducing A. hydrophila and the bacterial load in meatball. This study indicated that a dose of 0.75 kGy was sufficient to destroy approximately 10(4) cfu/g of A. hydrophila in meatball. PMID- 8717096 TI - [Detection of treatment of chicken breast with ionized rays and gradation of radiation dosage with the help of headspace gas chromatography and discriminant analysis evaluation]. AB - Chicken breast was irradiated with doses of 3, 5, and 7 kGy. Headspace gas chromatographical analysis demonstrated the tendency that the amounts of volatile compounds (mainly pentanal, hexanal and heptanal) are higher in irradiated samples in comparison with non irradiated. Statistical evaluation of the gas chromatograms by discriminant analysis enabled the detection of irradiation. Unknown samples could be classified in the groups "un-irradiated" or "irradiated" in most cases. PMID- 8717097 TI - Evaluation of the stable reaction products of histidine with formaldehyde or with other carbonyl compounds in dairy products. AB - Combined formaldehyde (FA) can be evaluated in cheese and other dairy products by determining spinacine (6-carboxy-1,2,3,4-tetrahydroimidazopyridine), an imino acid arising from the reaction of FA with the alpha-amino group of histidine. Other carbonyl compounds usually produced by fermentations can react themselves with histidine, forming molecules which interfere with the determination of spinacine. A sensitive (minimum detectable amount, 5 fmol spinacine) and interference-free HPLC method, with precolumn derivatization with o-phtalaldehyde (OPA) and 9-fluorenylmethylchloroformate (FMOC), to evaluate these histidine reaction-compounds in dairy products is described. The clean-up of the FMOC derivatized sample has been performed for the first time with solid-phase extraction (SPE) on an amino cartridge. The method was applied to samples of casein and differently ripened cheese, the origins of which were either known or commercial, in order to quantify the natural level of spinacine (0.7-2.7 ppm) probably deriving from biogenic FA, and to detect whether FA was used in processing. PMID- 8717098 TI - Synthesis of pyrraline reference material. AB - A simplified and improved method is described for the preparation of pyrraline, a lysine derivative from the advanced Maillard reaction and potential indicator for heat treatment of foods. The compound was obtained in a high degree of purity and with a yield of 31% from N alpha-t-butyloxycarbonyl-L-lysine after heating with 3 deoxy-D-erythro-hexos-2-ulose for 2 h at 70 degrees C in the dry state, preparative fractionation of the resulting N alpha-t-butyloxycarbonyl pyrraline with reverse-phase liquid chromatography and final deprotection of the intermediate compound with acetic acid. PMID- 8717099 TI - A new parameter for checking the suitability of alpha-tocopherol standards. AB - The quality of an alpha-tocopherol standard can be checked easily by measuring the UV absorbance at minimum (255 nm, Amin) and maximum (292 nm, Amax) wavelengths in n-hexane. If the quotient Amin/Amax exceeds 0.18, the standard contains less than 90% alpha-tocopherol and the determination at 292 nm will yield inaccurate results. PMID- 8717100 TI - Improvement in rheumatoid arthritis following application of an extracorporeal granulotrap column, G-1. AB - Patients with rheumatoid arthritis (RA) showed significantly (P < 0.01) increased numbers of granulocytes in their peripheral blood compared with normal donors and patients with osteoarthritis, and this finding correlated with interleukin-6 and C-reactive protein levels and active joint score. Then, 17 patients with RA were treated eight times in 4 weeks with a newly developed extracorporeal granulotrap column containing cellulose acetate beads (G-1 column). This column reduced granulocytes in the outflow blood by 50.2% compared with inflow counts. To evaluate the efficacy of G-1 therapy, 17 patients were followed for 12 weeks from the beginning of this therapy. The modified Lansbury index (LI) for monitoring RA activity significantly improved from a pretreatment mean score of 60.8% to a posttreatment score of 51.3%. The lowered scores were maintained up to 12 weeks after the initiation of therapy. Of the four LI items, tender and swollen joint scores showed the most significant improvement, with the tender joint score showing a particularly significant decrease throughout the study period. No serious side-effects were observed. These findings suggested that G-1 therapy was effective for RA. PMID- 8717101 TI - Quantification of morphometric changes in murine experimental osteoarthritis using image analysis. AB - The aim of this study was to evaluate a method for the quantification of cartilage erosions and osteophyte sizes in a murine model of osteoarthritis (OA). Mice in which OA was induced in the knee joint by intra-articular injection of bacterial collagenase were used. With an interactive image analysis system, the areas occupied by osteophytes and the areas of erosions of the articular cartilage were measured on histological sections by two independent observers at two time points. Measurements of osteophyte areas and cartilage loss at the tibial plateau showed good reproducibility, whereas measurement of cartilage loss at the femoral condyles was less reproducible. Measurement of three frontal total knee joint sections from the middle part of the joint provided a reliable measure for cartilage damage and osteophyte size in the total joint. A cumulative score was developed, composed of both cartilage loss and osteophyte size, which can be used as a general measure for OA of the knee joint. The presented method of quantitative scoring makes it possible to perform correlation studies and to investigate the effect of therapeutic interventions on the osteoarthritis process. PMID- 8717102 TI - IgM rheumatoid factor estimation by ELISA in seronegative rheumatoid arthritis before and after IgM fractionation: does seronegative RA exist? AB - We looked for the presence of IgM rheumatoid factor (RF) using an enzyme-linked immunosorbent assay (ELISA) in 25 patients with active seronegative rheumatoid arthritis. In unfractionated sera, 12 patients (48%) were positive for IgM RF (classical), but after IgM fractionation of 23 samples (2 samples were not available) using high performance liquid chromatography for fractionating IgM, 12 patients were positive for RF by ELISA indicating the presence of hidden RF. Finally, three patients were labelled as truly seronegative for IgM RF. Classical IgM RF as detected by ELISA correlated significantly with erosive disease. Hidden RF did not correlate with disease activity or severity in this cross-sectional study, and though its presence was associated with shorter disease duration this did not reach statistical significance. PMID- 8717103 TI - Long-term tolerability of methotrexate at doses exceeding 15 mg per week in rheumatoid arthritis. AB - The objective of this study was to examine longitudinally the tolerability of methotrexate (MTX) treatment at doses exceeding 15 mg/week in an open-label, prospective study. One hundred and eighty-five patients with rheumatoid arthritis were randomized to receive 15 mg or 25 mg MTX per week initially, and were followed over 30 months. Subsequent dose adjustments according to efficacy and tolerability resulted in levelling off of the mean dose at 18 mg/week, and the original treatment groups were combined for a longitudinal study comparing toxic events during months 1-12 and months 13-30. Withdrawals due to side-effects amounted to 17% during months 1-12 and 4% during months 13-30; dose reductions due to side-effects were 9% and 7%, respectively. The annual incidence of gastrointestinal side-effects increased from 26% to 39% (P = 0.05), that of liver enzyme elevation dropped from 43% to 10% (P < 0.001) and haemocytopenia remained stable at 5% and 7%. MTX pneumonitis was only observed during the first year, while airway complaints without evidence of parenchymal lung involvement increased to 10% beyond the first year. Fifty-six patients experienced 65 major infectious episodes over the 30-month period, with the respiratory tract being the most frequent site. This study showed that MTX treatment at doses exceeding 15 mg/week is tolerated over extended period of time. Major toxicity and withdrawals due to side-effects occurred predominantly during the first year of treatment and thus showed a decreasing trend over time, while minor toxic events continued throughout the study with a progressive rate of mucous membrane toxicity. MTX-treated RA appears to be a risk situation for major infection. PMID- 8717104 TI - The levels of memory (CD45RA-, RO+) CD4+ and CD8+ peripheral blood T-lymphocytes correlate with IgM rheumatoid factors in rheumatoid arthritis. AB - We investigated whether, in rheumatoid arthritis (RA), the CD45 isoform expression of peripheral blood T-lymphocytes (T-PBL) is related to auto-immune processes (e.g. IgM rheumatoid factors) and to clinical manifestations. By three colour flow cytometry, we quantified three subsets of CD4+ or CD8+ T-PBL: "naive" CD45RA+,RO-, "transient" CD45RA+,RO+, and "memory" CD45RA-,RO+ cells, in 102 patients with RA and in 41 age- and sex-matched controls. The serum levels of rheumatoid factors (RF) were determined--besides conventional agglutination tests -by ELISA (IgM-RF). Extensive clinical examination was performed at the time of blood sampling. In RA, age, sex and drug therapy did not constitute major influences on the CD45RA/RO patterns. In "healthy" men, higher age significantly' correlated with fewer naive and more memory CD4+ T-PBL (P < 0.01). In RA, distinct correlations between the T-PBL subsets, autoimmune and clinical manifestations became obvious when patients with low and high levels of RF against human IgG Fc fragments, as determined by ELISA, were analysed separately. RA patients with high IgM-RF had elevated proportions of CD45RO+ T-PBL (P < 0.05), that correlated with clinical parameters of disease activity (tender joint count, Ritchie index, P < 0.05) and outcome (Health Assessment Questionnaire, Larsen radiographic scores, P < 0.05). The proportions of memory CD4+ and CD8+ T PBL correlated strongly (P < 0.001) with the IgM-RF levels. Within 1 year, only three of 34 patients (disease duration of 5-9 years) showed seroconversion from low to high levels of IgM-RF (and positive agglutination tests); this was paralleled by reductions in naive and increases in transient T-PBL (P < 0.02). Thus, in RA, the proportions of memory CD4+ and CD8+ T-PBL correlate with the level of IgM-RF and, together with transient T-PBL, with clinical parameters of disease activity and outcome. PMID- 8717105 TI - Quantitative photometrical assessment of iron deposits in synovial membranes in different joint diseases. AB - We investigated 86 synovial membranes from patients suffering either from rheumatoid arthritis (RA) or osteoarthritis (OA). Iron deposits in the synovial membrane were stained by the Prussian blue reaction, and the amount of stained iron was quantitatively assessed by microscope photometry. We found a statistically significant increase in iron deposits in the synovial membrane of RA patients when compared to OA patients. The amount of iron deposits correlated with the histological subtype of synovitis, those presenting with more exudative and proliferative features showing greater amounts of iron deposits. We also observed an inverse correlation between the haemoglobin concentration and erythrocytes in the serum and the amount of iron in the synovial membrane. From our data we concluded that iron deposits in the synovial membrane can contribute by several mechanisms, including activation of oxygen radicals, to the chronic inflammatory reaction in RA synovitis. PMID- 8717106 TI - Finger skin temperature in patients affected by Raynaud's phenomenon with or without anticentromere antibody positivity. AB - We evaluated finger skin temperature in 36 patients affected by Raynaud's phenomenon with or without anticentromere antibody positivity. The temperature measured under basal conditions and after dipping the finger in water at 10 degrees C for 5 min was similar in both groups; the temperature at the end of an 18-min recovery period was significantly lower in anticentromere antibody positive patients. This observation suggests a persistent digital vasospasm after the cold test in patients suffering from Raynaud's phenomenon with anticentromere antibody positivity. This autoantibody seems to identify a subgroup of patients affected by Raynaud's phenomenon with a severe involvement of the microcirculation. PMID- 8717107 TI - [The status of aggressive children and victims of aggression in school]. AB - In a representative sample of 1594 pupils of the eight grade in Vienna and lower Austria the experiences of conflicts among pupils and the other school experiences were explored. Based on the reports of their own involvement in aggressive acts 4 groups of pupils were formed--a group which was themself aggressive against other pupils in their class but was no target of aggression (bullies); a group which was frequently a target of aggression; a group which was both--aggressive and target of aggression and finally an uninvolved group. It was shown that aggressive children had a high self-reported readiness to solve conflicts through physical power and that they were often involved in other dissocial acts in school. A major part of these pupils had a rather negative image of their school and of the teachers. The self-concept and the perception of their own role in class depended very much on their being a target of aggression or not. Parallels between these two subgroups and the distinction between socialized and undersocialized aggression are highlighted. In addition the special situation of those pupils who are a target of aggression without being aggressive is considered. PMID- 8717108 TI - [Treatment failures and analysis of resistance in behavior therapy exemplified by a parent-child program for the treatment of hyperkinetic and oppositional children]. AB - Resistance is a central concept of psychoanalysis, which is also increasingly regarded within behavior therapy. This article distincts resistance (an intervention is not accomplished) from failure (an intervention is accomplished but symptoms are not reduced) in behavior therapy. Using the example of a parent child-program for the treatment of hyperactive and oppositional children potential causes for failure and resistance in behavior therapy are demonstrated and the analysis of failure and resistance is described. The consideration of these problems of implementation are an important prerequisite for the improvement of the efficacy of behavioral interventions. PMID- 8717109 TI - [Documentation of treatment and quality assurance: recommendations for a practical solution]. AB - It is proposed to establish an internal quality assurance with a practical care documentation which at the same time will also fulfill the requirements of external quality assurance. A graphically economically designed questionnaire is presented. The integration of the diagnostic standards for child and adolescent psychiatry makes the questionnaire--made concrete for every clinic--usable in a communicably and dialogically external quality assurance. With the example of concrete data from the clinic of child and adolescent psychiatry in the hospital Bremen-East is shown how suitable indicators for quality are included in the care documentation. PMID- 8717111 TI - On multiple authorship. PMID- 8717110 TI - [The early intervention team--a model for multi-institutional cooperation in diagnosis of child developmental delay in a rural district]. AB - The author introduces the work of a multiprofessional recruted team, which diagnoses development disorders. It should be pointed out, that the local quality standard of diagnosing development disorders can be improved by intensifying the cooperation between the social institutions, which train or therapy children. Especially in the country far away from diagnostic centers this kind of cooperation can complete the psycho-social assistance network without calling for additional public recources. PMID- 8717112 TI - Imaging of muscle injuries. AB - Although skeletal muscle is the single largest tissue in the body, there is little written about it in the radiologic literature. Indirect muscle injuries, also called strains or tears, are common in athletics, and knowing the morphology and physiology of the muscle-tendon unit is the key to the understanding of these injuries. Eccentric muscle activation produces more tension within the muscle than when it is activated concentrically, making it more susceptible to tearing. Injuries involving the muscle belly tend to occur near the myotendinous junction. In adolescents, the weakest link in the muscle-tendon-bone complex is the apophysis. Traditionally, plain radiography has been the main diagnostic modality for evaluation of these injuries; however, with the advent of MRI it has become much easier to diagnose injuries primarily affecting the soft tissues. This article reviews the anatomy and physiology of the muscle-tendon unit as they relate to indirect muscle injuries. Examples of common muscle injuries are illustrated. PMID- 8717113 TI - The impact of osteonectin for differential diagnosis of osteogenic bone tumors: an immunohistochemical and in situ hybridization approach. AB - Thirty-three osteosarcomas at various grades of histologic differentiation, including chondroblastic, osteoblastic, and fibroblastic variants, were investigated immunohistochemically for evidence of osteonectin. Twenty-two cases of varying types of osteosarcoma were examined with in situ hybridization for mRNA expression of osteonectin. Immunohistochemically, osteonectin was present in all the osteosarcomas in this study. With in situ hybridization, 12 out of 22 osteosarcomas showed a positive signal. Two osteochondrosarcomas, seven chondrosarcomas, and one mesenchymal chondrosarcoma were also studied with regard to the localization of osteonectin, either immunohistochemically or by in situ hybridization. Immunohistochemically, osteonectin was present in all the chondroid lesions except for one osteochondroma. However, in situ hybridization of osteonectin mRNA was negative in all the chondroid lesions we studied. This study revealed that immunohistochemical localization of osteonectin is not useful in providing conclusive diagnosis of osteosarcoma. In situ hybridization of osteonectin mRNA might be useful in differentiating osteosarcoma from nonsteogenic bone tumors. PMID- 8717114 TI - Can conventional radiographs be used to monitor the effect of neoadjuvant chemotherapy in patients with osteogenic sarcoma? AB - OBJECTIVE: The objective of this study was to assess the effectiveness of conventional radiography in predicting histopathologic response in patients with osteogenic sarcoma who were treated with preoperative chemotherapy. DESIGN AND PATIENTS: The radiographs of 22 patients with an osteogenic sarcoma, taken before and after neoadjuvant chemotherapy, were reviewed. Tumour location, size, radiographic appearance, margination, cortical destruction and periosteal reaction were evaluated. The findings were correlated with the histopathologic response of the surgical specimen. RESULTS: None of the findings proved to be of predictive value for the histopathologic response. Increase in tumour diameter and increase in ossification and/or calcification, which were seen in more than half of the patients, did not correlate with response. CONCLUSION: Conventional radiographs do not contribute to the identification of good or poor responders. PMID- 8717115 TI - Benign fibrous histiocytoma of the femur: review of three cases. AB - OBJECTIVE: Benign fibrous histiocytoma is a tumor with histologic features similar to those of xanthofibromas, which include fibrous cortical defects and nonossifying fibromas, but with different clinical features. This study investigated the findings of benign fibrous histiocytoma on various imaging modalities. It emphasized the importance of clinical and imaging features as well as histologic findings in establishing the diagnosis. DESIGN: We reviewed three cases of benign fibrous histiocytoma of the femur that had been diagnosed histologically as (fibro)xanthoma during a retrospective analysis of 187 primary benign and malignant bone lesions. PATIENTS: All the patients were middle-aged women with various knee complaints. RESULTS AND CONCLUSIONS: Radiologically, the lesions were all lytic with well-defined geographic margins and sclerotic rims. The tumors arose within the medullary cavity in the distal metaphysis of the femur and involved the epiphysis. CT showed lytic destruction with well-defined marginal sclerosis. T1-weighted MR images showed that the lesions had predominantly the same signal intensity as that of skeletal muscle. The lesions were hypovascular on angiography. Bone scintigraphy showed moderately increased uptake. It was concluded that multiple images as well as clinical features can be useful in differentiating benign fibrous histiocytoma from fibroxanthoma, giant cell tumor, and other fibrohistiocytic tumors of bone. PMID- 8717116 TI - CT findings in osteoradionecrosis of the mandible. AB - CT scans of ten patients in whom the diagnosis of mandibular osteoradionecrosis was proven pathologically or by clinical follow-up were reviewed. All ten patients had bony abnormalities (cortical interruptions and loss of spongiosa trabeculation) on the symptomatic side. These were predominantly seen in the body of the mandible (premolar and molar region, eight patients), in some of these cases extending into the retromolar triangle (two patients) or mandibular angle (two patients). In the remaining two patients the abnormalities were in the ramus and angle. The two patients treated with iridium implantation showed localized lingual-sided cortical destruction. Three patients had a pathological fracture. The cortical destruction was buccal-sided in two and both buccal- and lingual sided in three of the other five patients. Contralateral bony abnormalities were present in four patients. Soft tissue thickening on the symptomatic side was seen in nine patients. As the bony abnormalities in mandibular osteoradionecrosis are often associated with a soft tissue mass, CT differentiation from tumor recurrence can be difficult. The association with cortical defects distant from the position of the original tumor (buccal surface or opposite side of mandible) should evoke the possibility of mandibular osteoradionecrosis. PMID- 8717117 TI - Glenoid hypoplasia and its relationship to instability. AB - Glenoid hypoplasia, or dysplasia of the scapular neck, is usually a bilateral and symmetric finding. Associated findings include hypoplasia of the humeral head, elongation of the glenoid cavity, and alterations in the shape of the coracoid or clavicle. Glenoid hypoplasia has been reported sporadically in the radiologic literature, but the condition may be more frequent than previously thought. In this paper, we discuss the radiographic findings of glenoid hypoplasia and explore the possibility of an association between glenoid hypoplasia and shoulder instability. PMID- 8717118 TI - Magnetic resonance imaging of dialysis-related amyloidosis of the shoulder and hip. AB - OBJECTIVE: The purpose of this study was to evaluate the usefulness of MRI in identifying abnormalities of the periarticular structures in patients with symptoms of dialysis-related amyloid arthropathy. DESIGN AND PATIENTS: MR images of shoulders and hips in 5 dialysis patients with symptoms of amyloid arthropathy were compared to images of shoulders and hips in 4 asymptomatic dialysis patients, shoulders in 9 nondialysis patients, and hips in 12 nondialysis patients. These were evaluated for the presence of focal periarticular osseous lesions, tendinous and capsular thickening, and periarticular fluid. RESULTS: Increased thickness of the supraspinatus tendon was found in both symptomatic and asymptomatic patients. Capsular thickening at the hip was present in all symptomatic patients, and in 3 of 8 asymptomatic hips. Joint and bursal fluid was common in the symptomatic group, and was present in some asymptomatic individuals. Osseous lesions were detected in the absence of plain film findings, and demonstrated variable signal intensity. CONCLUSIONS: MRI is useful in detecting signs of dialysis-related amyloidosis. Findings of a milder degree in asymptomatic dialysis patients suggests that MRI may also be valuable in the early diagnosis of this syndrome. PMID- 8717119 TI - Trabecular trauma of the talus and medial malleolus concurrent with lateral collateral ligamentous injuries of the ankle: evaluation with MR imaging. AB - OBJECTIVE: The objective of this study was to elucidate the relationship between lateral collateral ligamentous injuries of the ankle (ankle sprain) and bone bruise (trabecular trauma) of the talus and/or malleoli in the patients with twisting injuries of the ankle. DESIGN: Magnetic resonance studies of the ankle were retrospectively reviewed, focusing on the presence or absence of lateral collateral ligamentous injuries and the location of bone bruise in the talus and/or malleoli. PATIENTS: Thirty-five patients with acute twisting injuries of the ankle were studied. RESULTS AND CONCLUSION: Four patterns of bone bruise were found in 14 patients: (1) bone bruise in the talar dome equivalent to osteochondral fracture in two patients, (2) bone bruise in the posteromedial aspect of the talus and the medial malleolus singly or in combination in four, (3) bone bruise in the anteromedial aspect of the talus in six, and (4) bone bruise with a combination of the second and third patterns in two. The second pattern was associated with tear of the anterior talofibular (ATAF) ligament in all patients and injury of the calcaneofibular (CF) ligament in one out of four. The third and fourth patterns were associated with tears of both ATAF and CF ligaments in all patients. The identification of bone bruise was of value in indicating lateral collateral ligamentous injuries. PMID- 8717120 TI - Assessment of normal patellar cartilage volume and thickness using MRI: an analysis of currently available pulse sequences. AB - OBJECTIVE: The objective of this study was to analyse the potential of magnetic resonance imaging for valid determination of patellar cartilage thickness, comparing currently available pulse sequences. DESIGN: In six patients and one cadaver the cartilage was repetitively imaged employing three spin-echo and six three-dimensional gradient-echo sequences. In the cadaveric specimen the total volume and the regional distribution of cartilage thickness were assessed and compared with the values obtained from anatomical sections by image analysis. RESULTS AND CONCLUSIONS: The FLASH and fat-suppressed FLASH sequences allowed the most accurate determination of the cartilage volume and thickness. Fat suppression considerably increased the contrast of the cartilage to the synovial fluid, fat and bone marrow, yielding higher reproducibility of the volumetric measurements. The remaining difference from the anatomical volume and thickness may be because the calcified cartilage is not delineated by magnetic resonance imaging. PMID- 8717121 TI - MRI features in de Quervain's tenosynovitis of the wrist. AB - De Quervain's stenosing tenosynovitis of the first dorsal extensor component is traditionally diagnosed clinically but may be encountered when performing MRI of the wrist. A retrospective review of wrist MR images was performed in cases where the diagnosis of de Quervain's synovitis was suggested (n = 5). Imaging findings were correlated with clinical findings in four cases and with wrist arthroscopy in one case. Increased thickness of the extensor pollicus brevis and abductor pollicis longus tendons was the most reliable finding on MRI, being present in all cases. Peritendinous edema was also a reliable finding. Surrounding subcutaneous edema and increased intratendinous signal were less reliable findings in confirmed cases of de Quervain's disease. De Quervain's tenosynovitis may be encountered when performing MRI of the wrist. Increased tendon thickness and peritendinous edema are the most reliable imaging findings. PMID- 8717122 TI - A 16-month history of low back pain and spinal claudication symptoms. PMID- 8717123 TI - Giant bone island. PMID- 8717124 TI - Rickets in a 6-month-old monkey. AB - A 6-month-old male monkey with rickets was found dead by its keeper. Radiographic studies showed osteopenia of the entire skeletal system, a cup-shaped concavity and poor mineralization of metaphyseal ends, and markedly widened and irregular epiphyseal growth plates of the long bones and ribs. Gross anatomic findings included bowing abnormalities and microfractures in the radius, ulna, tibia, and fibia, and enlarged and widened cartilage columns extending into the metaphyses in the long bones and ribs. Histologic features included deficient mineralization and irregular disordered columns of proliferating cartilage in growth plates extending into other zones, dilated vascular channels, and poor mineralization of chondroid and osteoid tissues. PMID- 8717125 TI - Malignant lymphoma. PMID- 8717127 TI - Chondromyxoid fibroma. PMID- 8717126 TI - Aneurysmal bone cyst of metacarpal. PMID- 8717128 TI - Intraosseous lipoma of the distal femur: MRI appearance. PMID- 8717129 TI - Pseudotumor and arthropathy in the knees of a hemophiliac. PMID- 8717130 TI - Fibrous dysplasia with secondary aneurysmal bone cyst. PMID- 8717131 TI - Skeletal cystic angiomatosis. PMID- 8717132 TI - Malignant fibrous histiocytoma of the mandible. AB - We report a case of MFH of the mandible in a 16-year-old girl. The mandible is a rare location of MFH of bone, with only 21 cases described in the literature. Clinical, pathologic, and radiologic features are discussed, with special emphasis on the unusual pathologic and radiographic features. PMID- 8717133 TI - AIDS-related Kaposi sarcoma involving the tarsal bones. PMID- 8717134 TI - Chilling, oxidative stress and antioxidant responses in Arabidopsis thaliana callus. AB - Chilling of Arabidopsis thaliana (L.) Heynh. callus tissue to 4 degrees C led to conditions of oxidative stress, as indicated by increased levels of the products of peroxidative damage to cell membranes. Cellular H2O2 was also observed to increase initially upon chilling but by day 8 cellular levels had declined to below control levels. Although levels of catalase activity remained similar to those in unchilled tissue, activity of ascorbate peroxidase increased between days 4 and 8 of chilling to 4 degrees C. In callus held at 23 degrees C, levels of reduced glutathione remained static whereas they rose in callus held at 4 degrees C. Levels of oxidised glutathione were initially low but increased significantly by day 4 in the chilled callus. At 23 degrees C, however, levels of oxidised glutathione remained low. Between days 1 and 3 at 4 degrees C, levels of glutathione reductase activity increased but by day 8 glutathione reductase activity was similar to that in cells held at 23 degrees C. Exposure of callus to abscisic acid at 23 degrees C also led to increased activities of ascorbate peroxidase and glutathione reductase. PMID- 8717135 TI - Isolation of high-chlorophyll-fluorescence mutants of Arabidopsis thaliana and their characterisation by spectroscopy, immunoblotting and northern hybridisation. AB - Thirty-four recessive photosynthetic mutants of the high-chlorophyll-fluorescence (hcf) phenotype have been isolated by screening 7700 M2 progenies of ethyl methane sulfonate-treated seeds of Arabidopsis thaliana. Most of the mutants isolated were found to be seedling-lethal, but could be grown on sucrose supplemented media. Chlorophyll (Chl) fluorescence induction, absorption changes in the reaction-centre chlorophyll of PS I (P700) at 830 nm and Chl a/Chl b ratios were recorded in order to probe the photosynthetic functions and to define the mutational lesion. These studies were complemented by immunoblot and Northern analyses which finally led to the classification of the mutants into six different groups. Four classes of mutants were affected in PS I, PS II (two different classes) or the intersystem electron-transport chain, respectively. A fifth mutant class was of pleiotropic nature and the sixth class comprised a Chl b-deficient mutant. Several of the mutants showed severe deficiencies in the levels of subunits of PS I, PS II or the cytochrome b6/f complex. Thus the mutational lesions could be located precisely. Only one mutant was defective in the transcript patterns of some plastid-encoded photosynthesis genes. Hence most of the mutants isolated appear to be affected in translational and post translational regulatory processes of thylakoid membrane biogenesis or in structural genes encoding constituent subunits of the thylakoid protein complexes. PMID- 8717136 TI - Purification and characterization of glycosyltransferases involved in anthocyanin biosynthesis in cell-suspension cultures of Daucus carota L. AB - The major anthocyanins accumulated by an Afghan cultivar of Daucus carota L. are cyanidin 3-(xylosylglucosylgalactosides) acylated with sinapic or ferulic acid. The formation of the branched triglycoside present as a common structural element requires an ordered sequence of glycosylation events. Two of these enzymic glycosylation reactions have been detected in protein preparations from carrot cell-suspension cultures. The first step is a galactosyl transfer catalyzed by UDP-galactose: cyanidin galactosyltransferase (CGT) resulting in cyanidin 3 galactoside. The putative second step is the formation of cyanidin 3 (xylosylgalactoside) catalyzed by UDP-xylose: cyanidin 3-galactoside xylosyltransferase (CGXT). Both enzyme activities were characterized from crude protein preparations. The CGT was purified 526-fold from the cytosolic fraction of UV-irradiated cell cultures by ion-exchange chromatography on diethylaminoethyl (DEAE)-Sephacel, affinity chromatography on Blue Sepharose CL 6B, gel permeation chromatography on Sephadex G-75 and elution from the gel matrix after non-dissociating PAGE. Its molecular mass was estimated by SDS-PAGE and by calibrated gel permeation chromatography on Sephadex G-75. In both cases a molecular mass of 52 kDa was determined, indicating that the native protein is a monomer of 52 kDa. The galactosyl transfer and the xylosyl transfer are presumed to be catalyzed by separate enzymes. PMID- 8717137 TI - Abscisic acid causes changes in gene expression involved in the induction of the landform of the liverwort Riccia fluitans L. AB - The conversion of the submerged form of Riccia fluitans to the landform either by transfer to a moist solid surface or by treatment with abscisic acid (ABA), is accompanied by the formation of a set of new polypeptides and concomitant down regulation of other polypeptides. Changes in gene expression were analyzed by two dimensional separations of proteins and differential screening of a cDNA library. One of the landform-specific proteins might depend on the expression of the newly discovered Ric 1 gene. The deduced amino acid sequence of the isolated Ric 1 cDNA clone codes for a protein with a molecular mass of 30.1 kDa. This polypeptide possesses two amino acid sequences which are repeated five times each and it is largely hydrophilic with the exception of a hydrophobic carboxyl-terminal region. Under ABA treatment the expression of the Ric 1 mRNA had already reached its maximum after 1 h of incubation. Transferring submerged thalli onto an agar surface resulted in a slower induction. The Ric 1 gene product shows homology to an embryo-specific polypeptide of carrot seeds and to the group 3 of late embryogenesis-abundant (LEA) proteins. Interestingly, ABA treatment improved the desiccation tolerance of the submerged thalli. Additionally, ABA stimulated the synthesis of a protein which is immunologically related to a tonoplast protein. This finding, together with the fact that the ABA-induced landform exhibits an increased activity of several vacuolar enzymes, may indicate a special role of the tonoplast and the vacuole during ABA-induced conversion of the thallus from the submerged to the terrestrial form. PMID- 8717139 TI - Benzylaminopurine induces phenocopies of floral meristem and organ identity mutants in wild-type Arabidopsis plants. AB - Arabidopsis thaliana (L.) Heynh. has been used as a model system to investigate the regulatory genes that control and coordinate the determination, differentiation and morphogenesis of the floral meristem and floral organs. We show here that benzylaminopurine (BAP), a cytokinin, influences flower development in Arabidopsis and induces partial phenocopies of known floral homeotic mutants. Application of BAP to wild-type inflorescences at three developmental stages results in: (i) increase in floral organ number; (ii) formation of abnormal floral organs and (iii) induction of secondary floral buds in the axils of sepals. These abnormalities resemble the phenotypes of mutants, clv1 (increase in organ number), ap1, ap2, ap3 (abnormal floral organs) and ap1 (secondary floral buds in the axils of first-whorl organs). In addition, BAP induces secondary floral buds in the axils of perianth members of ap2-6, ap3-1 and ag mutants, and accentuates the phenotype of the ap2-1 mutant to resemble the ap2-6 mutant. These observations suggest that exogenous BAP suppresses the normal functioning of the genes for floral meristem identity and thereby affects flower development and the later stages of floral organ differentiation. PMID- 8717138 TI - Characterization of a novel drought-induced 34-kDa protein located in the thylakoids of Solanum tuberosum L. plants. AB - Using two-dimensional electrophoresis and Coomassie Blue staining, the accumulation of a 34-kDa protein (named cdsp 34 for chloroplastic drought-induced stress protein) is shown in the thylakoids of Solanum tuberosum plants subjected to a progressive and reversible water deficit. In-vivo labeling experiments showed an increased synthesis of cdsp 34 from the early stages of drought stress (leaf relative water content around 85%) and throughout the constraint. Sequences of the N-terminal part and of four tryptic-digest peptides did not reveal significant homology between the cdsp 34 protein and other known proteins. Western blotting analysis, using a serum raised against the N-terminal part of cdsp 34, confirmed the accumulation of cdsp 34 in thylakoids upon drought stress. From immunoblot analysis of different chloroplastic subfractions, the cdsp 34 protein appears to be an extrinsic protein preferentially located in unstacked stroma thylakoids. Immunoprecipitation of in-vitro-translated products, as well as Southern analysis, showed that the cdsp 34 protein is nuclear encoded. After rewatering of water-stressed plants, the level of cdsp 34 synthesis was reduced, but remained substantially higher than in control plants. Western analysis showed the persistence of a high amount of cdsp 34 in rewatered plants for at least two weeks. Based on the abundance and on the location of cdsp 34 within thylakoids, a putative role for this novel chloroplastic protein is discussed in relation to the tolerance of the photosynthetic apparatus of higher plants to dehydration. PMID- 8717140 TI - [Isovolemic hemodilution in coronary heart disease--clinical and hemodynamic effects]. AB - Clinical and hemodynamic effects of isovolemic hemodilution (HD) were evaluated in 12 patients (aged 59 +/- 8 years) with severe multivessel coronary artery disease (CAD) and angina pectoris grade III (Canadian Cardiovascular Society classification) despite high-dose medical treatment. In none of these patients was aortocoronary bypass grafting or percutaneous transluminal coronary angioplasty possible. Prior to HD and after 3 months of HD the incidence of angina pectoris was determined by means of questionnaires; hemodynamic measurements were performed with right heart catheterization at rest and during exercise. After 3 months of HD hematocrit was reduced from 46.2 +/- 1.3% to 38.5 +/- 0.5%. The weekly incidence of angina pectoris was unchanged (19 +/- 7 before, 17 +/- 8 after HD). Cardiac index was 2.5 +/- 0.7 1/min/m2 at rest and 3.9 +/- 1.0 1/min/m2 during exercise before, 2.6 +/- 0.5 1/min/m2 at rest and 3.9 +/- 0.8 1/min/m2 during exercise after HD. Stroke volume index did not increase significantly neither at rest nor during exercise after HD. Initially, systemic vascular resistance decreased from 1659 +/- 603 to 1398 +/- 420 dyns/cm5 during exercise; after HD it was 1522 +/- 551 (rest) and 1283 +/- 348 dyns/cm5 (exercise). Mean pulmonary artery pressure (PAP) and wedge pressure (WP) were unchanged at rest (PAP: 19.9 +/- 6.7 mm Hg before, 19.2 +/- 6.5 mm Hg after HD; WP: 10.8 +/- 5.5 mm Hg before, 10.7 +/- 4.3 mm Hg after HD) and during exercise (PAP: 43.0 +/- 9.9 mm Hg before, 42.8 +/- 8.9 mm Hg after HD; WP: 30.8 +/- 4.6 mm Hg before, 30.6 +/- 6.5 mm Hg after HD). In conclusion, in patients with CAD isovolemic HD does not reduce angina pectoris but also does not induce clinical deterioration. Furthermore, isovolemic HD does not worsen the hemodynamic effects of severe CAD with impaired left ventricular function. PMID- 8717141 TI - [Coronary heart disease and impaired left ventricular function in patients over 80 years of age--is bypass operation justified?]. AB - Between April 1988 and February 1995, 35 patients aged 80 years or older underwent elective coronary artery bypass grafting. Concomitant aortic valve replacement was performed in eight patients. Fifteen patients with moderate to severe left ventricular dysfunction (left ventricular ejection fraction (EF) 20 to 45%) (group I) were compared to 20 patients with an EF > 45% (group II). There were no differences according to age, NYHA classification and associated diseases. Group I patients had significantly more myocardial infarctions compared to group II (1.3 +/- 1.1 vs. 0.5 +/- 0.8; p < 0.05). Also more bypass grafts per patient were used in group I (3.1 +/- 0.9 vs. 2.3 +/- 0.9; p < 0.01). No differences between the groups were observed in the postoperative course. One patient of group II died from multiorgan failure (hospital mortality 2.2%). A follow-up could be performed in 32 of 34 hospital survivors (94%) 12.0 +/- 11.2 months (1-41 months) postoperatively. There was one non-cardiac death in each group. All hospital survivors improved by at least one NYHA functional class. Actuarial survival at 3 years is 91% in both groups. In patients aged 80 years or older with or without left ventricular dysfunction coronary artery bypass surgery can be performed with an acceptable risk and a functional benefit. Thus, elective operations should be performed in this age group despite left ventricular dysfunction. PMID- 8717142 TI - [Anomalous origin of the left coronary artery from the right sinus of Valsalva with acute symptoms]. AB - The case of a 52-year-old man with an anomalous origin of the left coronary artery from the right sinus of valsalva coursing between the aorta and pulmonary trunk is reported with an acute myocardial ischemia. He had an additional high grade main-stem stenosis in the course behind the pulmonary artery. The anatomical variants, clinical significance, and cause of ischemia are discussed. Also, technical considerations in both diagnosis and management are given. PMID- 8717143 TI - [Diastolic left ventricular dysfunction with microvascular angina (Syndrome X)]. AB - Findings on left ventricular function in microvascular angina (syndrome X) are somewhat controversial. Recently, an increased prevalence of insulin resistance and hyperlipoproteinemia has been demonstrated as well as arterial hypertension potentially impairing the left ventricular diastolic function. In an attempt to analyze the diastolic function at rest, we investigated by Doppler echocardiography the transmitral blood flow in 16 patients (51 +/- 8 years). The diastolic data were compared with those of 12 healthy people (50 +/- 6 years) who were matched for age. The patients with microvascular angina revealed a slightly higher systolic blood pressure (134 +/- 18 mm Hg vs 125 +/- 9 mm Hg, n.s.), but imposed by higher left ventricular mass index (92 +/- 18 g/m2 vs 65 +/- 13 g/m2, p < 0.001). There was a trend to an elevated A-wave-peak during transmitral blood flow (61 +/- 14 cm/s vs 50 +/- 9 cm/s). The findings suggest impairment of the left ventricular relaxation before manifestation of left ventricular hypertrophy and hypertension takes place. PMID- 8717144 TI - [Dobutamine stress echocardiography and perfusion scintigraphy for detection of myocardial ischemia in patients with and without previous myocardial infarct]. AB - Dobutamine stress echocardiography has proven to be a method with high diagnostic accuracy in the detection of coronary artery disease. In case of previous myocardial infarction it is of importance to detect additional regions with inducible myocardial ischemia. This study aimed at the detection of inducible ischemia by dobutamine stress echocardiography and stress perfusion scintigraphy in patients without and with previous myocardial infarction. 50 patients without as well as 50 patients with previous transmural myocardial infarction were investigated. In all patients coronary angiography, technetium-99m methoxy isobutyl-isonitrile (MIBI)-SPECT after bicycle ergometry and dobutamine stress echocardiography (up to 40 mcg/kg/min dobutamine, 1 mg atropine) were performed within 14 days. In patients with previous myocardial infarction dobutamine stress echocardiography and MIBI-SPECT had similar sensitivities (91 vs. 94%, n.s.) and specificities (81 vs. 75%; n.s.) in the detection of significant coronary artery disease. Agreement on the presence or absence of inducible ischemia was 84% (Kappa = 0.60). In patients with previous transmural myocardial infarction sensitivity of stress echocardiography and perfusion scintigraphy in the detection of significant coronary artery disease is lower with 63% and 77%, respectively. In this patient group transient perfusion defects were found more frequently than inducible wall motion abnormalities, 76% and 60%, respectively. There was a lower agreement (76%; Kappa = 0.49) in the detection of abnormal or normal results between dobutamine echocardiography and stress perfusion scintigraphy for this group of patients. CONCLUSION: This study demonstrates high agreement of dobutamine stress echocardiography and stress perfusion scintigraphy in the evaluation of inducible ischemia in patients without previous transmural myocardial infarction and equal diagnostic accuracy in the detection of coronary artery disease. In patients with previous myocardial infarction there is a lower agreement in the interpretation of patients as having ischemia due to negative dobutamine echo results in patients having positive perfusion scintigraphies. PMID- 8717145 TI - [Feasibility and safety of dobutamine stress echocardiography: experiences with 1,000 studies]. AB - The use of dobutamine stress echocardiography for the evaluation of coronary artery disease is rapidly expanding. Despite its widespread use, the feasibility and safety of dobutamine stress echocardiography has not been sufficiently documented. Between November 1992 and June 1995, we performed 1000 dobutamine stress echocardiographies. There were 744 men and 256 women with a mean age of 59 +/- 11 years. Anti anginal medication was not routinely withdrawn before the test. The mean maximal dobutamine dose was 41,4 +/- 10 mu g/kg center dot min( 1). Atropine was given additionally in 440 patients, with a mean dose of 0.5 mg. In patients receiving beta-blockers additional atropine was more often necessary as compared to those not receiving beta-blockers (278/457 = 61% versus 162/543 = 30 %, p < 0.0001). Reasons for discontinuing dobutamine infusion were achievement of target heart rate (64 % of cases) and maximal dose (12 % of cases). In 791 (79,1 %) patients no side-effects of dobutamine stress echocardiography were noticed. Termination of the study because of adverse side-effects occurred in 6.6 %. A total of 103 (10,3 %) noncardiac side-effects were observed: dizziness or nausea 6.4 %, headache 1.7 %. In one patient a focal cerebral seizure occurred. 156 cardiac side-effects occurred: blood pressure decrease of more than 20 mm Hg in 25 patients, extreme palpitations in 16 patients and pulmonary edema in one case. Most common cardiac side-effects consisted of arrhythmias (11.4 %): 9.1 % ventricular and 2.3 % supraventricular arrhythmias. Most ventricular arrhythmias were less severe (uniform and multiform premature ventricular beats, ventricular bigeminy or couplets in 71 patients). Nonsustained ventricular tachycardia, with a maximum duration of 20 s, occurred in 18 patients. In one patient sustained ventricular tachycardia developed and progressed towards ventricular fibrillation. This patient could be successfully defibrillated. Supraventricular arrhythmias presented as new atrial fibrillation in 10 patients, supraventricular tachycardia in three patients, junctional rhythm with a short decline in heart rate in nine patients and a second-degree AV block in another case. Dobutamine stress echocardiography has proven to be a safe and feasible method in the diagnosis of coronary heart disease. Minor side-effects are common and sometimes unpleasant for the patient, but do not often require termination of the study. Severe side-effects are seldom (< 1 %), but nevertheless, adequate medical and technical (defibrillator) support should be rapidly available. PMID- 8717146 TI - [2D and Doppler echocardiography after atrial switch operation for transposition of great arteries; a review]. AB - Introduction of the atrial baffle procedure (Mustard, Senning) more than 30 years ago has dramatically improved the prognosis of patients with transposition of the great arteries. However, a range of late complications can occur even in clinically asymptomatic patients including: superior and inferior limb obstruction, pulmonary venous obstruction, residual baffle-leakage, left ventricular outflow tract obstruction, tricuspid valve regurgitation, right ventricular dysfunction and postoperative dysrhythmia. As an increasing number of patients has reached adulthood an accurate noninvasive technique is needed for routine follow-up studies. In most patients transthoracic two-dimensional (2D) and Doppler echocardiography allows a complete investigation of postoperative hemodynamic abnormalities -- in particular assessment of atrial baffle function. In adult patients with restricted echo-windows transoesophageal echocardiography may provide a more detailed assessment of atrial baffle morphology and function; TEE seems to be superior compared to angiography and magnetic resonance imaging. Furthermore TEE is a reliable guiding and monitoring technique during interventional cardiac catheterization procedures such as balloon dilatation or stent implantation and allows further surveillance. Even if compared to other noninvasive diagnostic procedures, 2D and Doppler echocardiography (transthoracic/transoesophageal) are still the most important and accurate diagnostic methods for routine investigation of the postoperative anatomy and potential late complications in patients who underwent an atrial baffle procedure. PMID- 8717147 TI - [Color Doppler echocardiography of the flow convergence region in vitro: effect of the orifice shape on proximal velocity profile]. AB - The flow convergence method serves to determine flow across orifices (like valve leaks) by color Doppler. Both the PISA method (proximal isovelocity surface areas) and the PVP method (proximal velocity profile) were developed in vitro at circular orifice plates. Therefore, we studied the influence of a non-circular orifice shape on the color map of the flow convergence. Steady flow across orifices of the following shapes was imaged by color Doppler: Oval (6 x 2 mm), slit (12 x 1.5 mm), three-star (diameter 100, area 30 mm2), circular twin-orifice (two circular orifices diameter 2 mm at 10 mm distance from each other) and oval twin-orifice (two ovals 6 x 2 mm at 10 mm distance). As reference we imaged circular orifices with a similar opening area. The alias method was used to locate discrete velocities within the color map, and the proximal velocity profile along the flow center line was analyzed (mean of 24 subsequent images). The local velocity was plotted (y-axis) against its distance to the orifice (x axis) providing proximal velocity profile curves. The more the orifice shape differed from circular, the more the proximal velocity profile was shifted downward: The profile proximal to the oval was not different from the reference profile proximal to the circular orifice. The profile proximal to the slit was considerably slowed, and proximal to the three-star was even slightly slower (local velocity -12 %, -23 % and -29 % at 14, 8 and 5 mm distance to the orifice). If the circular reference orifice corresponded to total flow across the twin-orifice, the proximal velocity profile of the latter was also shifted markedly downward (-20 %, -18 % and -23 % at 14, 8 and 5 mm distance to the circular twin-orifice). However, if the reference profile corresponded to flow across only one opening of the twin-orifice, the proximal velocity profile of the latter was shifted considerably upwards (+60 %, +71 % and +50 % at 14, 8, and 5 mm distance). Deviation of the orifice shape from circular leads to lower local velocities within the flow convergence; thus neglecting this orifice shape would result in underestimation of flow by the flow convergence method. However, presence of parallel neighboring flow increases the local velocities; neglecting this effect would lead to corresponding overestimation of flow. PMID- 8717148 TI - [Form fruste of long QT syndrome]. AB - The long QT syndrome (LQTS) is an inherited disorder associated with recurrent syncope and sudden death from ventricular arrhythmias. It is characterized by a prolonged QT interval on the ECG. A case is discussed in which a patient presented with recurrent syncopal episodes on exertion associated with a slightly prolonged QT interval. Family history revealed an increased incidence of sudden deaths in two generations. Some reports on similar patients suggest that there may be a forme fruste of the long QT syndrome. The additional diagnostic values of exercise stress test, Valsalva maneuver and genetic testing are evaluated. After analysis of the patient's and mother's ECGs the diagnosis of a forme fruste of LQTS is made. This case is presented to emphasize the importance of diagnosis of borderline cases of LQTS because of the high risk of sudden death for untreated patients. PMID- 8717149 TI - [Atrial flutter in young patients after corrective operation of congenital heart defects: therapy by high frequency catheter ablation]. AB - Radiofrequency catheter ablation has been used successfully in adult patients for treatment of atrial flutter. Three young patients (mean age 14.6 years) with common type atrial flutter (n = 2) and uncommon type atrial flutter (n = 1) underwent electrophysiologic study. Cardiac diagnoses included tricuspid atresia after Fontan operation, status after closure of an atrial septal defect of secundum type, and status after surgical valvotomy for valvular pulmonary stenosis, respectively. Indication for ablation were syncopal episodes in one and presyncopal episodes in two patients. Flutter cycle length ranged from 280 to 320 ms. Right atrial endocardial mapping revealed areas with local electrograms preceding the onset of the flutter-P-wave by 70 to 80 ms. In these regions pace mapping was performed with the intention to produce an identical P-wave morphology and short stimulus to P-wave interval. Radiofrequency current application (500 kHz) with 30 W for 30 s at theses sites terminated atrial flutter and prevented reinduction in all three patients. No complications were observed. Follow-up (7 to 11 months) revealed recurrence of uncommon type of atrial flutter in one patient who previously had had common type. The remaining two patients are in stable sinus rhythm. CONCLUSION: Radiofrequency catheter ablation appears to be an effective treatment of atrial flutter after surgery for congenital heart defects in young patients. PMID- 8717150 TI - [Heart transplantation: after-care and rehabilitation. Current status and results of a survey by the German Transplantation Program. Thoracic Organ Transplantation Study Group of the German Society of Cardiology]. PMID- 8717151 TI - Stimulation of histamine H2- (and H1)-receptors activates Ca2+ influx in all trans-retinoic acid-differentiated HL-60 cells independently of phospholipase C or adenylyl cyclase. AB - In human neutrophils, histamine H2-receptors mediate activation of adenylyl cyclase (AC) and inhibition of N-formyl-L-methionyl-L-leucyl-L-phenylalanine (FMLP)-induced superoxide anion (O2-) formation, and in HL-60 promyelocytes, H2 receptors mediate parallel activation of AC, phospholipase C (PLC) and non selective cation (NSC) channels. As all-trans-retinoic acid (RA) is successfully used in the differentiation therapy of acute promyelocytic leukaemia, we studied signal transduction in RA-differentiated HL-60 cells. Histamine and the H2 receptor agonist, impromidine, induced both rises in cAMP levels and cytosolic Ca2+ ([Ca2+]i). Substances acting at post-receptor sites to increase cAMP did not increase [Ca2+]i. H2- but not H1-receptor antagonists inhibited histamine-induced cAMP accumulation and rises in [Ca2+]i were more effectively inhibited by H2- than by H1-receptor antagonists. Histamine-induced rises in [Ca2+]i were completely dependent on the presence of extracellular Ca2+ and were abolished by the blocker of NSC channels, Gd3+, but were resistant to inhibition by pertussis toxin. Unlike FMLP, histamine did not activate PLC. The effects of FMLP on [Ca2+]i were less sensitive to blockade by Gd3+ than those of histamine, and there was no cross-desensitization between the two stimuli. FMLP, but not histamine, inhibited transiently thapsigargin-induced rises in [Ca2+]. Taken together, our results show that histamine activates AC-mediated cAMP accumulation in RA-differentiated HL-60 cells via H2-receptors and NSC channel-mediated Ca2+ influx via H2- (and H1)-receptors. Histamine-induced NSC channel activation is not the consequence of AC- or PLC stimulation and occurs, directly or indirectly, via pertussis toxin-insensitive guanine nucleotide-binding proteins. FMLP and histamine activate Ca2+ influx by different mechanisms. There are similarities in H2-receptor-mediated signal transduction between RA-differentiated HL-60 cells and HL-60 promyelocytes and differences between the former cells and neutrophils, indicating that RA-differentiated HL-60 cells must be considered as partially immature. PMID- 8717152 TI - The effect of endothelin-2 (ET-2) on migration and changes in cytosolic free calcium of neutrophils. AB - The effect of endothelin-2 (ET-2) on neutrophil migration and intracellular calcium was studied. Depending on the concentration, ET-2 enhanced or inhibited neutrophil migration. At low concentrations ET-2 caused a chemotactic stimulation of migration, in contrast with endothelin-1 (ET-1) which caused a chemokinetic stimulation of migration. At higher concentrations ET-2 inhibited formyl methionyl-leucyl-phenylalanine (fMLP)-activated migration. Both activation and inhibition by ET-2 were completely dependent on extracellular Ca2+. Unlike ET-1 which caused an increase in cytosolic free Ca2+ at a concentration which stimulated migration, ET-2 caused a measurable increase of cytosolic free Ca2+ at a concentration which did not stimulate migration. This strongly suggests that there is no correlation between maximal stimulation of cytoplasmic free calcium, and maximal stimulation of migration. Influx of extracellular Ca2+ was required for both activation of migration and change in cytosolic free Ca2+, because no effect was observed in the absence of extracellular Ca2+, and because blockers of Ca(2+)-influx inhibited ET-2-activated migration. The ETA-receptor antagonist cyclo-(-D-Trp-D-Asp-Pro-D-Val-Leu) (BQ123), and the ETB-receptor antagonist [Cys11-Cys15]-endothelin-1(11-21) (IRL1038) antagonized the stimulatory effect of ET-2 on migration, and the inhibitory effect of high concentrations of ET-2 on fMLP-activated chemotaxis. This suggests that both the ETA-receptor and the ETB receptor are involved in the stimulatory effect of low concentrations of ET-2, and in the inhibitory effect of high concentrations of ET-2. PMID- 8717153 TI - Differential effects of organotin compounds on voltage-gated potassium currents in lymphocytes and neuroblastoma cells. AB - Effects of organotin compounds were studied on voltage-gated K+ current in whole cell voltage clamped lymphocytes and in N1E-115 neuroblastoma cells. In human peripheral blood lymphocytes the immunotoxic compounds dibutyltinchloride (DBT, 2.5 microM) and triphenyltinchloride (TPhT, 2.5 microM) decrease the peak amplitude of the K+ current and prolong time to peak. Tributyltinchloride (TBT, 2.5 microM) decreases the K+ current to a greater extent than DBT and TPhT, without affecting the time to peak. The neurotoxic organotin compound trimethyltinchloride (TMT, 2.5 microM) does not affect the voltage-gated K+ current in lymphocytes. Similar effects of DBT were observed in freshly isolated and PHA-activated human lymphocytes and with rat thymocytes. On the other hand, in mouse N1E-115 neuroblastoma cells, none of the organotin compounds altered the voltage-dependent K+ current. In human lymphocytes DBT affects both the peak amplitude and the time to peak of the K+ current in a concentration-dependent manner. At the maximum concentration of 10 microM tested, the peak amplitude of the K+ current was reduced to 22 +/- 4% of the control current. The IC50 and slope factor for block of the peak outward current by DBT amounts to 6.7 +/- 0.4 microM, and 2.7 +/- 0.4, respectively. The delay in K+ current activation does not saturate. At 10 microM DMT increases the time to peak to 332 +/- 12% of the control value. The present results suggest that the effects by DBT originate from two separate interactions with the voltage-gated K+ channel at the extracellular site of the membrane: a direct effect on the closed K+ channel causing a delay in current activation and a membrane-related effect causing inhibition of the K+ current. The differential effects of the organotin compounds may relate to their differential toxicological action. PMID- 8717154 TI - Pharmacology of cloned human 5-HT1D receptor-mediated functional responses in stably transfected rat C6-glial cell lines: further evidence differentiating human 5-HT1D and 5-HT1B receptors. AB - This study was undertaken to investigate the pharmacology of human serotonin (5 HT)1D receptor sites by measuring two functional cellular responses, inhibition of forskolin-stimulated cAMP formation and promotion of cell growth, using transfected rat C6-glial cell lines and a broad series of 5-HT receptor agonists. Stable and separate transfection of a pcDNA3 or pRcRSV plasmid, each containing a cloned human 5-HT1D receptor gene, in rat C6-glial cells was confirmed with RT PCR of 5-HT1D receptor mRNA and radioligand binding with [3H] 5 carboxamidotryptamine (5-CT) and [3H] sumatriptan. The 5-HT1D receptor density was 350 and 1050 fmol/mg protein for the C6-glial/pcDNA3/5-HT1D and C6 glial/pRcRSV/5-HT1D cell line, and forskolin (100 microM)-induced cAMP formation was inhibited by 45 and 78% in the presence of 1 microM 5-HT, respectively. A comparison of the intrinsic agonist activities for sixteen 5-HT receptor ligands with their corresponding binding affinities for the human 5-HT1D receptor site showed similar results for both cell lines with the exception of the partial agonist m-trifluoro-phenyl-piperazine (TFMPP). Three classes of compounds were observed: 1) efficacious agonists, such as 5-CT, 5-methoxytryptamine, 5-HT, sumatriptan, bufotenine, 5-methoxy-3(1,2,3,6-tetrahydro-4-pyridinyl)1H-indole (RU 24,969), tryptamine and 8-hydroxy-2(di-n-propilamino)tetralin (8-OH-DPAT), with agonist potency close to their binding affinity; 2) the partial agonists metergoline, 7-trifluoromethyl-4(4-methyl-1-piperazinyl)-pyrolo-(1,2-a) quinoxaline (CGS 12066B), 1-naphthylpiperazine and 2'-methyl-4-(5-methyl [1,2,4]oxadiazol-3-yl)-biphenyl-4-carboxylic acid [4-methoxy-3-(4-methylpiperazin 1-yl)-phenyl]-amide (GR 127,935) with marked intrinsic agonist activity but at concentrations higher than their binding affinity; and 3) the silent antagonists ritanserin, ketanserin and methiothepin, apparently free of intrinsic agonist activity, with antagonist potency close to their binding affinity. The cAMP data were further supported by the observed promotion of cell growth by stimulation of both transfected cell lines with sumatriptan under serum-free conditions; half maximal stimulation was obtained at 4.4 nM (C6-glial/pcDNA3/5-HT1D) fully in agreement with its EC50-value (5.7 nM) for inhibition of cAMP formation. This growth promoting effect was antagonised by 1 microM methiothepin and not observed in pcDNA3-plasmid-transfected and non-transfected C6-glial cells. A comparative study with a C6-glial/pcDNA3/5-HT1B cell line expressing a similar amount of cloned human 5-HT1B receptors (Bmax: 360 fmol/mg protein) showed almost no intrinsic agonist activity for metergoline, 1-naphtylpiperazine and GR 127,935. Together with the 5-HT1D receptor binding selectivity and antagonist activity of ketanserin and ritanserin, the findings define important pharmacological differences between cloned human 5-HT1D and 5-HT1B receptor sites. PMID- 8717155 TI - Hypothalamic paraventricular nucleus involvement in the pressor response to N methyl-d-aspartic acid in the periaqueductal grey matter. AB - The aim of this study was to demonstrate paraventricular hypothalamic nucleus (PVN) involvement in the cardiovascular changes induced by N-methyl-d-aspartic acid (NMDA) microinjections at the level of periaqueductal grey (PAG) matter. The study was carried out in anaesthetized rats and the arterial blood pressure monitored by a polygraph. NMDA injections (0.68-6.8 nmol/rat) into the PAG area induced a significant increase in blood pressure. After pretreatment by injection of the NMDA receptor antagonist dl-2-amino-5-phosphonovaleric acid (2-APV, 0.05-5 nmol/rat) into the PVN, administration of NMDA (0.68 nmol/rat) into the PAG area elicited a decrease, rather than an increase, of blood pressure. We observed a significant reduction of the pressor effect induced by 6.8 nmol/rat NMDA after 2 APV injection into the PVN. 2-APV injection into the dorsomedial hypothalamic nucleus, an area near the PVN, did not modify the increase in blood pressure induced by NMDA in the PAG area. We suggest the existence of a glutamatergic connection between the PAG area and the PVN in the cardiovascular effects of NMDA. PMID- 8717157 TI - Behaviour-related effects of nicotine on slow EEG waves in basal nucleus-lesioned rats. AB - The basal magnocellular nucleus is assumed to play a crucial role in cholinergic activation of the cortical EEG. The aim of this study was to establish whether intraperitoneally applied nicotine may counteract the power asymmetry of the slow waves in the cortical EEG of both hemispheres after an unilateral lesion in the basal nucleus. In 17 rats the basal nucleus (substantia innominata/ventral pallidum) was unilaterally lesioned by ibotenic acid. The lesion produced unilateral power increases of all frequencies up to 20 Hz in the frontal EEG that increased with higher arousal level. Additionally, synchronized spike and wave discharges appeared in the frontal EEG. The results indicate that the basal nucleus suppresses especially the delta EEG waves in the frontal motor cortex during motor active behaviour. Nicotine (0.1 and 1 mg/kg) partially counteracts the power asymmetry of frontal slow waves (2-6 Hz) only during exploratory sniffing but not during grooming and waking immobility. Physostigmine (1 mg/kg) was also effective during exploratory sniffing. The results may indicate a role of nicotinic mechanisms in the information input component of exploratory behaviour. PMID- 8717156 TI - Altered in vivo catecholamine release in the hypothalamic paraventricular nucleus of the aged rat. AB - The effect of age on basal and stimulated noradrenaline release in the hypothalamic paraventricular nucleus (PVN) of the rat was examined by in vivo microdialysis. Microdialysis probes were inserted into the PVN of 3 and 18 month old anaesthetised Sprague Dawley rats and perfused with a modified Ringer solution. Following four basal 30-min collections, transmitter release was stimulated by perfusion with 100 mM potassium for one collection. After re equilibration, blood pressure was raised 60 mmHg for 30 min by phenylephrine infusion (1-1.3 mg/kg) then a 2-h recovery period followed. Dialysate collections were injected directly onto a reverse phase HPLC-ECD (HPLC with electrochemical detection). Basal extracellular noradrenaline concentrations were found to be similar in adult and old animals. Basal dihydroxyphenylacetic acid (DOPAC) concentrations were significantly greater in old compared to adult rats (P < 0.05). Potassium depolarisation induced a significant increase in noradrenaline concentrations in both age groups (P < 0.001), however the noradrenaline response to potassium stimulation was significantly reduced in the aged rats (P < 0.05). Potassium-induced decreases in DOPAC and homovanillic acid (HVA) concentrations were seen in both age groups. Following phenylephrine infusion, a modest delayed reduction in noradrenaline levels, which failed to reach statistical significance, was seen. Phenylephrine-induced hypertension was associated with decreased DOPAC and HVA concentrations in adult (P < 0.05) and old (P < 0.05) rats, respectively. These results indicate that ageing is associated with changes in dopaminergic and noradrenergic activity in the PVN of the rat. A reduction in noradrenaline response to maximal stimulation induced by potassium depolarisation was observed with ageing. The alteration in the activity of the catecholaminergic pathways to the PVN induced by phenylephrine infusion appears to be age dependent. PMID- 8717158 TI - Failure of tyramine to release neuronal ATP as a cotransmitter of noradrenaline in the guinea-pig vas deferens. AB - Contractions, release of noradrenaline and release of ATP elicited by the indirectly acting sympathomimetic amine tyramine and responses elicited by exogenous noradrenaline were studied in the isolated vas deferens of the guinea pig. Release of noradrenaline was assessed as overflow of tritium after preincubation with [3H]-noradrenaline. ATP was measured by means of the luciferin luciferase technique. In tissues pretreated with pargyline 1 mM, tyramine 300 microM, when added to the superfusion medium for 2 min, elicited contraction and an overflow of tritium (mainly [3H]-noradrenaline) and ATP. Contraction and ATP overflow responses were prevented and tritium overflow was greatly reduced by desipramine 10 microM. Prazosin 0.3 microM abolished contractions and evoked ATP overflow without changing tritium overflow. Blockade of postjunctional P2 purinoceptors by suramin 300 microM caused a marked decrease of tyramine-evoked contractions and a slight reduction of tritium overflow whereas evoked ATP overflow was markedly increased. The effect on contraction was not shared by two other P2-purinoceptor antagonists, namely pyridoxalphosphate-6-azophenyl-2',4' disulfonic acid (PPADS) 32 microM and diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS) 32 microM: PPADS increased contractions about fourfold, whilst DIDS had no effect at all. When the vas deferens was superfused for 24 min with medium containing tyramine 300 microM, evoked contractions and tritium overflow continued throughout whereas ATP overflow faded rapidly to basal values. In the presence of prazosin 0.3 microM, tyramine 300 microM again failed to elicit contractions as well as an overflow of ATP. Application of noradrenaline 10 microM instead of tyramine also resulted in prolonged contraction and an overflow of ATP that declined rapidly. It is concluded that all ATP released by tyramine is non-neuronal in origin, secondary to the activation of postjunctional alpha 1 adrenoceptors by released noradrenaline. The non-neural ATP does not seem to play a functional role in smooth muscle contraction and derives from a postjunctional source which is subject to a rapid depletion upon sustained alpha 1-adrenoceptor activation. PMID- 8717159 TI - Opposite modulation of cotransmitter release in guinea-pig vas deferens: increase of noradrenaline and decrease of ATP release by activation of prejunctional beta adrenoceptors. AB - Effects of isoprenaline on contraction, release of noradrenaline and release of ATP elicited by electrical field stimulation (210 pulses, 7 Hz) as well as on contractions elicited by exogenous noradrenaline and ATP were studied in the isolated vas deferens of the guinea pig. Release of noradrenaline was assessed as overflow of total tritium after preincubation with [3H]-noradrenaline. ATP was measured by means of the luciferin-luciferase technique. In [3H]-noradrenaline pretreated tissues, electrical stimulation elicited an overflow of tritium and ATP and a biphasic contraction. Isoprenaline (1-100 nM) reduced the contraction, mainly phase I, and enhanced the evoked overflow of tritium: evoked overflow of ATP was not changed significantly. No, or almost no, contraction remained in [3H] noradrenaline-pretreated tissues exposed to both prazosin (0.3 microM) and suramin (300 microM), and the evoked overflow of ATP was reduced by about 82%. Under these conditions, isoprenaline (1-100 nM) again enhanced the evoked overflow of tritium, but it now decreased the evoked overflow of ATP. Propranolol (1 microM), when added on top of prazosin and suramin, prevented the effects of isoprenaline (1-100 nM). In some tissues not pretreated with [3H]-noradrenaline, purinergic and adrenergic components of the neurogenic contraction (again to 210 pulses, 7 Hz) were isolated by exposure to prazosin (0.3 microM) and suramin (300 microM), respectively. Isoprenaline (1-100 nM) decreased the isolated purinergic component but did not change significantly the isolated adrenergic component. Contractions elicited by ATP (1000 microM) were not changed and contractions elicited by noradrenaline (100 microM) were slightly increased by isoprenaline (1 100 nM). Isoprenaline (100 nM) did not change the degradation of ATP (100 microM) by pieces of the vas deferens. It is concluded that, in the guinea-pig vas deferens, activation of prejunctional beta-adrenoceptors modulates the neural release of noradrenaline and ATP in opposite directions: release of noradrenaline is enhanced, whereas release of ATP is decreased. PMID- 8717160 TI - The contribution by monoamine oxidase and catechol-O-methyltransferase to the total-body and pulmonary plasma clearance of catecholamines. AB - To study the effects of inhibition of catechol-O-methyltransferase (COMT) and monoamine oxidase (MAO) on the removal of circulating catecholamines, anaesthetized rabbits were infused for 120 min with 3H-labelled noradrenaline, adrenaline and dopamine. Total-body plasma clearances (Cltot) and pulmonary fractional extractions (ERp) of the infused amines and the cardiac output of plasma (CO(p)) were determined under steady-state conditions at the end of each of two consecutive 60-min treatment periods. MAO and COMT were inhibited by treatment with pargyline (40 mg/kg) and tolcapone (3 mg/kg followed by 1.5 mg/kg given every 30 min), respectively. Two groups of animals were studied. Group I involved animals treated with tolcapone throughout and given pargyline at the beginning of the second treatment period. In group II, pargyline was given at the beginning of the first, and the treatment with tolcapone was started at the beginning of the second treatment period. As previous experiments had shown that COMT inhibition alone is without any effect on Cltot of the three catecholamines considered here, the results obtained in the first treatment period of group I can be taken to reflect control results. At the end of the first treatment period, Cltot of noradrenaline, adrenaline and dopamine (expressed as a percentage of CO(p)) was 88%, 85% and 142%, respectively, in group I (COMT inhibition) and 67%, 77% and 115%, respectively, in group II (MAO inhibition; P < 0.05 for the group difference regarding Cltot of noradrenaline and dopamine). MAO inhibition on top of COMT inhibition (group I) lowered Cltot of noradrenaline, adrenaline and dopamine by 23%, 12% and 26%, respectively, and COMT inhibition on top of MAO inhibition (group II) reduced Cltot of these catecholamines by 13%, 20% and 17%, respectively. At the end of the first treatment period, the pulmonary plasma clearance (Clp = ERp x CO(p)) of noradrenaline and dopamine was 13 and 25 ml kg-1 min-1, respectively, in group I and 12 and 28 ml kg-1 min-1, respectively, in group II. Clp of adrenaline did not differ from zero in either group. Clp of noradrenaline and dopamine was reduced by 74% and 70%, respectively, when both enzymes were inhibited in group I and by 70% and 67%, respectively, when both enzymes were inhibited in group II. Hence, inhibition of either MAO or COMT alone had little, if any, effect on the removal of noradrenaline, adrenaline and dopamine on passage through the systemic and pulmonary circulation. Combined inhibition of both MAO and COMT was highly effective in reducing the pulmonary clearance of noradrenaline and dopamine, but produced only minor decreases in the total-body clearance of all three catecholamines. PMID- 8717161 TI - Effects of selective phosphodiesterase inhibitors on platelet-activating factor- and antigen-induced airway hyperreactivity, eosinophil accumulation, and microvascular leakage in guinea pigs. AB - There is currently interest in the potential use of selective inhibitors of cyclic nucleotide phosphodiesterases (PDE) in the treatment of asthma. In this study we examined the effects of three selective PDE inhibitors, milrinone (PDE III), rolipram (PDE IV) and zaprinast (PDE V), on the broncoconstriction produced by antigen and histamine, the airway hyperreactivity and microvascular leakage after aerosol exposure to platelet-activating factor (PAF) and antigen, and the antigen-induced eosinophil infiltration in guinea-pig lung. Inhaled rolipram (0.01-10 mg ml-1) inhibited dose dependently the bronchospasm produced by aerosol antigen (5 mg ml-1) an anaesthetised, ventilated guinea-pigs. Rolipram (10 mg ml 1) produced maximal inhibition of antigen-induced bronchoconstriction but only partial inhibition of the response to aerosol histamine (1 mg ml-1). Milrinone and zaprinast (each 10 mg ml-1) showed weak, or no, inhibitory effects against bronchoconstriction produced by aerosol antigen or histamine. Pretreatment with rolipram (10 mg kg-1, i.p.) prevented airway hyperreactivity to histamine which develops 24 h after exposure of conscious guinea-pigs to aerosol PAF (500 micrograms ml-1) or antigen (5 mg ml-1). The pulmonary eosinophil infiltration obtained with 24 h of antigen-exposure was inhibited by rolipram. In contrast, milrinone and zaprinast (each 10 mg kg-1, i.p.) failed to reduce either the airway hyperreactivity of the eosinophil accumulation in these animals. Rolipram (1-10 mg ml-1) reduced the extravasation of Evans blue after aerosol PAF (500 micrograms ml-1) at all airway levels while a lower dose (0.1 mg ml-1) was only effective at intrapulmonary airways. Rolipram (0.01-1 mg ml-1) markedly reduced airway extravasation produced by inhaled antigen (5 mg ml-1). Zaprinast (1-10 mg ml-1) was also effective against airway microvascular leakage produced by aerosol PAF or antigen while milrinone (10 mg ml-1) had no antiexudative effect. These data support previous suggestions that pharmacological inhibition of PDE IV results in anti-spasmogenic and anti-inflammatory effects in the airways and may be useful in the treatment of asthma. PMID- 8717162 TI - Expression of cytochrome P 450 3A enzymes in human lung: a combined RT-PCR and immunohistochemical analysis of normal tissue and lung tumours. AB - We have previously demonstrated expression of cytochrome P 450 3A (CYP3A) protein in pulmonary carcinomas and surrounding normal tissue, using immunohistochemistry. These results suggested that different CYP3A enzymes may be expressed in normal and tumour tissue. Therefore, the aim of the present study was to identify specific CYP3A enzymes expressed in normal human lung and lung tumours. Both normal lung tissue and tumour tissue from eight patients was analyzed for CYP3A4, CYP3A5 and CYP3A7 mRNA using a specific RT-PCR (reverse transcriptase-polymerase chain reaction) method. Identical samples were subjected to immunohistochemical analysis of CYP3A protein. CYP3A5 was the major enzyme of the CYP3A subfamily present at the mRNA level in both normal human lung and lung tumours. CYP3A5 mRNA was detected in normal lung tissue in all eight cases and in tumour tissue in four cases. CYP3A7 mRNA was detected in five cases in normal tissue and in one tumour. Notably, no CYP3A4 mRNA was found in any of the samples. Immunohistochemical staining for CYP3A protein was found in normal lung tissue in each case. Interestingly, all pulmonary carcinomas showed immunostaining for CYP3A, while mRNA for CYP3A enzymes was found in only four cases. In summary, our study indicates a specific expression pattern of the members of the CYP3A subfamily in normal human lung and lung tumours. These findings have potential clinical significance, since it has been recently shown that CYP3A5 catalyzes the activation of the anticancer pro-drugs cyclophosphamide and ifosfamide. Thus, local activation of these agents may take place in pulmonary carcinomas and surrounding normal tissues. PMID- 8717163 TI - Chronic (-)-isoprenaline infusion down-regulates beta 1- and beta 2-adrenoceptors but does not transregulate muscarinic cholinoceptors in rat heart. AB - Regulation of beta-adrenoceptor (beta-ar) subtypes and transregulation of muscarinic cholinoceptors (mAchr) was examined in regions of rat heart after chronic infusion of (-)-isoprenaline (450 micrograms/kg per hour) for 14 days. Following (-)-isoprenaline infusion systolic blood pressure was reduced for 10 days but then gradually returned to control levels, whereas heart rate was increased for 7 days before declining to a level significantly above control. Heart weight to body weight ratio was increased in (-)-isoprenaline treated rats. beta-ar subtype densities were measured by quantitative autoradiography with [125I]-cyanopindolol (CYP) in sinoatrial node (SA), atrioventricular node (AV), bundle of His (BH), left (LB) and right (RB) bundle branches, interventricular (IVS) and interatrial (IAS) septa, right atria (RA), apex (AX) and mitral valve (MV). beta 1-ars were reduced by 59.1-74.2% in the AV conducting regions, 53.4% in the SA node and 43.3-53.4% in myocardial areas, beta 2-ars were markedly reduced in myocardial regions (93.2-98.5%) and in pacemaker and conducting regions (87.7-97.8%). No changes in mAchr densities measured using [3H]-N-methyl scopolamine (NMS) occurred in the AV node, BH, LB, RB, IVS and IAS following (-) isoprenaline infusion. Densities of beta 1- and beta 2-ars and mAchrs were also measured in ventricular homogenates from control and (-)-isoprenaline treated animals. beta-ar levels were significantly reduced (P < 0.05) in treated animals and the ratio of beta 1- to beta 2-ars increased after treatment. mAchr density in ventricular homogenates measured using either [3H]-NMS or [3H]-quinuclidinyl [phenyl-4-3H]benzilate (QNB) was unchanged. Homogenates of left and right ventricle also showed no change using [3H]-NMS. Organ bath studies were used to investigate the effect of (-)-isoprenaline infusion on negative inotropic and chronotropic effects of the non-selective muscarinic receptor agonist bethanechol in left and right atria, respectively. Lower concentrations of bethanechol (3 x 10(-10) to 10(-6) M) produced a negative inotropic response in isolated electrically driven left atria from (-)-isoprenaline treated rats, but not from control rats, with the slope of the curves being significantly different between groups (ANCOVA, P = 0.037). At concentrations of bethanechol from 10(-6) to 3 x 10(-4) M the negative inotropic response was not changed between (-)-isoprenaline treated and control animals. Bethanechol also produced a negative chronotropic response at lower concentrations (10(-10) to 10(-6) M) in (-)-isoprenaline treated rats, but not in controls. A second, steeper phase of the negative chronotropic response occurred at concentrations of bethanechol greater than 10( 6) M and was also seen in control rats. Expression of M2 (cardiac) mAchrs (m2Achr) in left and right ventricular tissues measured using a quantitative non competitive polymerase chain reaction (PCR) assay showed a significant (P = 0.001) 28.5% increase in expression in left ventricle and a significant (P = 0.003) 21.5% decrease in expression in right ventricle after (-)-isoprenaline treatment, compared to controls. There was no significant difference in total ventricular m2Achr expression between the two groups of rats. The results suggest that chronic beta-ar stimulation down-regulates both beta 1- and beta 2-ars, and appears to differentially transregulate m2Achr expression, but not mAchr protein. Following (-)-isoprenaline infusion, muscarinic receptor mediated responses were sensitised, with no change in receptor densities, suggesting changes occur in the cell signalling system beyond the level of the receptor. PMID- 8717164 TI - Effects of the class III antiarrhythmic drug ambasilide on outward currents in human atrial myocytes. AB - We have studied the inhibitory influence of the class III antiarrhythmic drug ambasilide (LU 47110) on the transient outward current Ito1 and the sustained current Iso following inactivation of Ito1, in human atrial myocytes. The two currents are separated by a mathematical procedure based on the amplitudes and time constants of the biexponential inactivation of the total outward current. The frequency dependence, the recovery from inactivation and the kinetics of activation and inactivation are described. Ambasilide reversibly and concentration dependently inhibited Ito1, Iso and the sodium current INa. Concentration required for half maximal inhibition (IC50) for the effects on Ito1 and Iso were 23.3 mumol/l and 45.7 mumol/l respectively, concentrations shown by others to be effective in terminating and preventing fibrillation in a dog atrial arrhythmia model. Ambasilide not only reduced the amplitude of Ito1 and Iso but also accelerated the time course of inactivation from 14.22 to 6.69 ms and from 202.3 to 87.9 ms respectively. The amplitude of Ito1 showed only a small dependence on stimulation frequency characteristic for human atrial myocytes, whereas Iso was reduced significantly at higher stimulation frequencies. Ambasilide did not change these relationships (0.1-4 Hz) and therefore did not show the reverse use-dependence known from other class III antiarrhythmic agents and which is an important property for a prospective antiarrhythmic drug. The lack of an effect of ambasilide on both steady-state activation and inactivation of Ito1, and the time constant of recovery from inactivation, suggests that ambasilide acts by changing conductance rather than by influencing the gating mechanism. The described characteristics make ambasilide an interesting substance in the group of class III antiarrhythmic drugs. PMID- 8717165 TI - RPR 101821, a new potent cholesterol-lowering agent: inhibition of squalene synthase and 7-dehydrocholesterol reductase. AB - RPR 101821 (trans-2-[4-(benzoxazol-2-yl)-phenylmethoxy] amino cyclohexane hydrochloride) is a potent cholesterol-lowering agent in rodents and marmoset. The compound inhibited rat liver microsomal squalene synthase (IC50 = 1 nM) and 7 dehydrocholesterol (7DHC) reductase (IC50 = 1 microM; Lewis et al. 1995). When RPR 101821 (10 mg/kg), the 7DHC reductase inhibitor BM 15.766 (4[2-[4-(4 chlorocinnamyl)piperazine-1-yl]ethyl] benzoic acid; 10 mg/kg) or the HMG-CoA reductase inhibitor lovastatin (30 mg/kg) was given orally to rats at -29 h, -21 h and -5 h, serum cholesterol was reduced by 56%, 46% or 15%, respectively. The reduction in cholesterol with RPR 101821 was associated with an accumulation of 7DHC in serum, suggesting an inhibition of 7DHC reductase. In the presence of BM 15.766, RPR 101821 reduced the serum accumulation of 7DHC in a dose-dependent manner, with complete inhibition at 30 mg/kg, p.o. In Balb-cJ mice, RPR 101821 and lovastatin (50 mg/kg, b.i.d., p.o., for 14 days) lowered serum cholesterol by 67% and 2%, respectively. In marmosets, RPR 101821 and lovastatin (both at a dose of 10 mg/kg, p.o., b.i.d., for 7 days) reduced cholesterol by 28% and 19%, respectively. In summary, RPR 101821 is an orally effective potent cholesterol lowering agent in rodents and a small primate species. The suggested mechanism of hypocholesterolemic effect is the inhibition of squalene synthase and 7DHC reductase. PMID- 8717166 TI - Cardiac ATP-sensitive K+ channel: a target for diadenosine 5',5''-P1,P5 pentaphosphate. AB - In numerous studies the intracellular mononucleotide-dependent gating of ATP sensitive K+ (KATP) channels has been demonstrated. However, it is not known whether dinucleotide polyphosphates, a family of endogenous compounds structurally-related to ATP, could also modulate this ion conductance. Therefore, in the present study we assessed the direct effect of diadenosine 5',5''-P1,P5 pentaphosphate (Ap5A) on cardiac KATP channel activity using the inside-out configuration of the patch-clamp technique. Addition of Ap5A (50 microM) to the internal side of membrane patches, excised from guinea-pig ventricular cells, strongly inhibited KATP channel activity. The estimated NPO (where N is the number of channels in the patch and PO the open probability of each channel) was 4.16 +/- 0.50 in the absence and 0.85 +/- 0.30 in the presence of Ap5A (50 microM). This effect of Ap5A was partially reversible, and the NP0 was 2.26 +/- 0.60 after washout of Ap5A. Exposure of KATP channels to increasing concentrations of Ap5A revealed that the Ap5A-induced inhibition is concentration dependent with the half-maximal effective concentration of 16 microM (Hill coefficient: 1.6). On the basis of these results, we conclude that Ap5A is a potent antagonist of the KATP channel activity. This represents a previously unrecognized property of Ap5A, as well as the discovery of a potentially novel endogenous ligand of myocardial KATP channels. PMID- 8717167 TI - [Behavior of the lactate level in occlusion and reperfusion of the right superior mesenteric artery. An animal experiment study]. AB - The value of lactate in the early clinical diagnosis of acute mesenteric arterial occlusion was evaluated in an animal model. The superior mesenteric artery of pigs (8-9 weeks old, 20-23 kg) was clamped for 1 h (group 2, n = 9) and 3 h (group 3, n = 6). Reperfusion was recorded for 2 h. Follow-up measurements were done in six animals (group 1). The systolic blood pressure was elevated significantly to about 18-40 mmHg in the initial phase of ischemia (P = 0.02). The intramucosal pHi measured tonometrically was decreased to below a value of 7.08 (P = 0.03), and the flow of superior mesenteric vein was reduced by about 90% ( P = 0.03) in groups 2 and 3. Within 30 min the lactate increased to about 2.05-3.8 mmol/l in the central venous blood sample and 2.8-4.8 mmol/l in the portal vein blood sample in 30 min. After 3 h of ischemia (group 3) the elevated lactate level returned to normal without any significant difference. In the reperfusion period (group 2) the systolic blood pressure stabilized at a depressed level (63-73 mmHg, P = 0.0054), and the flow of the superior mesenteric vein showed a reduction of 41% (P = 0.03). Intramucosal pHi and lactate values returned nearly completely to normal within 2 h. Reperfusion after 3 h of ischemia (group 3) caused marked shock without any sign of recovery at intramucosal pH or mesenteric vein flow. The lactate values increased for above the ischemic level. In conclusion, an elevated lactate level only correlates with the initial phase of acute mesenteric arterial occlusion. A normal lactate concentration cannot exclude the diagnosis of mesenteric ischemia. Reperfusion of the ischemic intestine is characterized by circulatory shock with secondary increased lactate concentration without predictive value for adequate intestinal perfusion. PMID- 8717168 TI - Ruptured hepatic angiolipoma in tuberous sclerosis complex. AB - Tuberous sclerosis complex (TSC) may be associated with hepatic angiomyolipoma. In contradistinction to renal angiomyolipoma this lesion is rare, and nontraumatic hemorrhage has not so far been described. This is the first report of a spontaneously ruptured, surgically treated and histologically verified, hepatic angiomyolipoma in TSC. Anomalies of the vessel walls and consequently the hazard of bleeding are known to increase with the diameter of the tumor in other localizations. It is concluded that in cases with rapid tumor growth and marked vessel anomalies elective surgery should be considered. PMID- 8717169 TI - [Alloplastic augmented femoral reinserted anterior cruciate liganent. Value of 2 channel augmentation for postoperative rehabilitation]. AB - The rupture of the anterior cruciate ligament (ACL) near its femoral origin is a common injury of the knee and can lead to lesions of the meniscus due to instability and to early gonarthrosis. One procedure applied in current orthopaedic practice to prevent such impairment of knee joint function is ACL repair reinforced with a synthetic intraarticular ligament. In this study we used twelve knees of cadavers and after sectioning the ACL in each repaired it according the Marshall technique with USP 1 PDS II sutures. We augmented the repair in each case with a 3-mm PET (Trevira hochfest) band inserted by the through-the-condyle (TTC) procedure and attached without preload to the femoral and tibial condyle with a 4-mm staple. We then measured the length of the ACL, the length of the Marshall sutures-ACL complex, the partial lengths, and the deviation angles and adherence-friction force of the 3-mm PET augmentation device, and applied the law of Hooke to calculate the load-sharing between the USP 1 PDS II sutures-ACL complex and the 3-mm PET band and between the ACL and the 3-mm PET band, respectively. We also evaluated the load on the femoral and the tibial fixation of the augmentation device. The results showed that the 3-mm PET band took over 60% of an externally applied load on the knee during the hypothetic period of ACL healing and 27% of the force acting on the knee thereafter. It was calculated that a maximum of 75% of the load taken over by the augmentation device was at the tibial staple and only up to 45% of the force at the femoral one. With due consideration for the requirement for absolute protection ("stress shielding") of the healing ACL but also for the aim of early postoperative accelerated functional rehabilitation without casts, splints or other restrictions of joint movement, we believe that a pretensioned 3-mm PET band is the best choice, since augmentation without preload cannot fulfil these requirements. PMID- 8717170 TI - [Therapy of carcinoids of the stomach]. AB - Carcinoids of the stomach are rare but have gained importance since the introduction of acid secretion inhibitors. The most important type is the tumor occurring in patients with chronic atrophic gastritis with and without pernicious anemia. The tumors are benign and can be treated by local endoscopic or surgical methods. Antrectomy reduces hypergastrinemia and may cause regression of the tumor. Sporadic carcinoids of the gastric antrum are malignant, however, and require radical surgical treatment. PMID- 8717172 TI - [Endoguide Reinbek--a device for trocarless insertion of instruments in endoscopic surgery]. AB - The Endoguide Reinbek (EGR) is a device that allows the insertion and use of laparoscopic instruments without a trocar-sheath unit. It consists of a steel rod 40 cm long and 10 mm in diameter, with pointed ends that can be screwed off. One end is 10 mm in diameter and the other, 6 mm. Both points can be covered by a removable handle. APPLICATION: The EGR is introduced into the abdominal cavity via a 10-mm port. Camera supervision is applied to select the exact position for any instrument. The tip of the EGR perforates the abdominal wall from inside, thus preventing any damage to blood vessels of viscera. A small skin incision is made over the tip once the skin bulges outward at the body surface. Once outside the abdomen the tip is removed, leaving a socket into which any instrument, e.g. a forceps, fits. The instrument is "guided" into the abdominal cavity by simply pushing it axially to the EGR. Inside the abdomen the EGR is disconnected from the instrument and withdrawn. The instrument is used without a port. There is no more loss of gas than when trocars are used. The EGR is especially useful for instruments that do not have to be changed during the operation and for the application of endoscopic drains. However, instruments can also be changed. The main advantage of the EGR is cost reduction, especially if disposable trocars have been used thus far. If reusable trocars have been applied so far there is a significant reduction in the maintenance effort (dismanting, cleaning, reassembly and resterilization of trocars). Moreover, endoscopic instruments can be positioned more safely and more precisely. PMID- 8717171 TI - [Malignant tracheal tumors--surgical experiences in 6 patients with primary malignancies of the trachea. Current diagnosis and therapy]. AB - In a retrospective study six patients with malignant tumours of the trachea located distal to the cricoid and proximal to the carina tracheae were operated on during the period of December 1983 to July 1995 in the Department of Thoracic Surgery in Hannover (Heidehaus). Histopathological examination revealed two adenoid cystic carcinomas, two squamous cell carcinomas, one mucoepidermoid carcinoma, and one low-grade sarcoma. Laser bronchoscopic resection was performed in one patient and tracheotomy one other avoid imminent asphyxia. Physical examination mainly showed a range of symptoms such as stridor and dyspnoea; spirography showed increased airway resistance, and X-ray of the trachea was important in the diagnostic process. The tracheal tumours were diagnosed by histopathological examination of excised material obtained by bronchoscopy. Three patients underwent resection and primary reconstruction of the trachea, with a length of resection between 2.0 and 3.5 cm and end-to-end anastomosis. Endotracheal afterloading was necessary in the case of one female patient with tumour infiltration of the proximal end of the upper trachea. Neoadjuvant irradiation was followed by resection of the whole trachea and implantation of a tracheal prosthesis (Neville) in the case of a patient with extensive endo- and extraluminal tumour infiltration. An extensive recurrence led to the death of this patient 5 months after the surgical intervention. Because of the distal location, resection of the tracheal bifurcation was necessary in two patients, in one combined with a pneumonectomy on the right side. Stenosis of the main bronchus and development of granuloma made stent implantation unavoidable. Dislocation of the stents and recurrent pneumonia caused the patients deaths 74 days and 18 months postoperatively. On follow up 17, 93, and 120 months postoperatively none of the other patients had recurrences. Current diagnostic and therapeutic options for malignant tracheal tumours are discussed. PMID- 8717173 TI - [Immune paralysis in acute pancreatitis--HLA-DR antigen expression on CD14+DR+ monocytes]. AB - Determination of the prognosis in acute cases of pancreatitis, particularly in its serious and necrotizing form, still presents problems. Patients require intensive care and suffer from severe septic complications that do not correlate with pancreatic enzyme levels (amylase lipase). METHOD: Thirty-one patients with acute pancreatitis were examined: group 1 -- necrotizing pancreatitis (lethal outcome n = 7); group 2 -- necrotizing pancreatitis (surviving n = 12); group 3 edematous pancreatitis (surviving n = 12). For 11 consecutive days after admission to a clinical ward, flow cytometric check-ups were carried out daily on all patients. The antigen-presenting system HLA-DR antigen expression on monocytes and C-reactive protein were examined. RESULTS: When groups 1 and 2 were compared with group 3, HLA-DR values on monocytes were significantly different following the third day after admission (P < 0.01). Comparison of groups 1 and 2 were significant from the third day of observation (P < 0.001). During all 11 days of observation, patients in group 1 remained in immune paralysis (HLA-DR expression on monocytes CD14+DR+20% antigen density). All of these patients had infected necroses. Patients in group 2 overcame their immune paralysis. HLA-DR depression of monocytes and a long-standing high C-reactive protein level are almost certain predictors of a fatal outcome in cases with severe pancreatitis. A routine passage cytometric check/FACS to determine the activity of monocytes (HLA DR) is of prognostic significance. PMID- 8717174 TI - [Long-term parenteral nutrition in vascular-induced short bowel syndrome]. AB - Mesenteric vascular occlusion with intestinal infarction is often regarded as a fatal illness. Often the diagnosis of severe ischemia is made during laparotomy. Sometimes resection is not performed because the conditions is thought to be incurable, and in such cases the mortality is 100%. If radical and aggressive resection is carried out even at the site of almost complete small bowel infarction and followed by an elective second-look operation, survival can be achieved in some patients. The motivation to attempt treatment should be further enhanced by the observation that the short-bowel syndrome following resection can be successfully treated, giving an acceptable quality of life for the patients. Between 1 January 1979 and 31 December 1979 we treated 9 male and 6 female patients with short-bowel syndrome after mesenteric occlusive disease; their ages ranged between 4 and 78 years, and the total duration of parenteral nutrition was 30,494 patient-days. With a median of 1503 days, the longest individual periods of parenteral nutrition were 4,919 and 5,015 days. We show the problems of long term parenteral nutrition, including catheter-associated and nutritional complications, with reference to the individual courses of patients with short bowel syndrome after mesenteric infarction. Despite some problems, patients of all ages can return to near-normality with an acceptable quality of life with the aid of parenteral nutrition at home. PMID- 8717175 TI - [Results of surgical therapy in renal hyperparathyroidism. Follow-up of 143 patients]. AB - Subtotal parathyroidectomy (SPTX) and total parathyroidectomy with autotransplantation (PTX and AT) are now equivalent and challenging surgical procedures, that now involve very low morbidity even in patients with serious secondary illness. In most cases, either brings about a significant amelioration of quality of life. It is rare for the illness to persist after such an operation. In most cases, this is the case only after an inadequate primary operation. An actual recurrence of the illness after at least 6 months is a much more serious problem and cannot usually be attributed to an inadequate operation. Our experience is based on operations performed in 143 patients and a follow-up of up to 8 years. In 68.7% of cases there was a noticeable improvement of bone pain immediately after the operation. This rate fell to 61.5% during subsequent years. Comparable results could not be achieved with any drug therapy. After SPTX the recurrence rate was 4.5%, and after PTX and AT, 5.5%. This is comparable to the results cited in the literature. PMID- 8717176 TI - [Tumor cell dissemination in bone marrow and peritoneal cavity. An immunocytochemical study of patients with stomach or colorectal carcinoma]. AB - The tumor spread and the radicality of surgical resection are the most important facts in a patient's prognosis. In spite of curative tumor resection many patients die from metastases or local tumor recurrence. One possible reason is early dissemination of tumor cells which cannot be detected with clinical methods of examination. For this reason the aim of our study was to examine both bone marrow and peritoneal lavage for disseminated tumor cells with an immunocytochemical technique in patients with a gastrointestinal carcinoma. We also wanted to find out whether there was any correlation between the incidence of tumor cell detection and the TNM classification, staging and tumor grading and whether disseminated tumor cells have any prognostic significance. Our study included 54 patients who underwent surgery in our clinic for a carcinoma of the stomach (20 patients) or the colorectum (34 patients) from November 1993 to December 1994. At the beginning of the operation bone marrow had been taken from the iliac spine, and the abdomen was irrigated with 1000 ml saline solution immediately after laparotomy or laparoscopy. After cell separation with Ficoll density centrifugation 5 x 10(5) cells were applied per slide by a cytospin technique. For detection of the tumor cells we used the APAAP technique and the following monoclonal antibodies: KL1, CK2, anti-CEA, 17-1A (bone marrow) and Ber EP4, B72.3, anti-CEA and 17-1A (peritoneal lavage). Altogether 77% of all patients had tumor cells in the bone marrow and 69% in peritoneal lavage fluid. It was possible to detect tumor cells in bone marrow (67%) and peritoneal lavage fluid (25%) even of patients with T1 tumors. The percentage increased with depth of wall infiltration. There was a marked difference in bone marrow aspirates between patients with lymph-node-negative tumors (N0) and those with lymph-node positive tumors (N+): 65% had tumor cells in N0 and 85% in N+ stages. This trend was also seen in patients with (M1) and without (M0) metastases, in both bone marrow aspirates and peritoneal lavage fluid. In bone marrow there was a good correlation of tumor cells with staging, but in peritoneal lavage fluid this was not so. Finally, we detected tumor cells more often in bone marrow and peritoneal lavage fluid of patients with poorly differentiated tumors (G3) or diffuse Lauren type than in patients with moderately differentiated tumors (G2) or intestinal Lauren type. After a median follow-up period of 12.5 months patients with disseminated tumor cells had a lower survival rate than patients without tumor cells. PMID- 8717177 TI - [Possibilities and limits of scores. Theoretical considerations about scores]. AB - Scores are intended to facilitate an accurate appreciation of the overall situation from a few characteristic, reasonably weighted parameters. This definite result in the form of a score means a reduction of reality. For the individual it cannot reasonably be applied in practice. Despite a long list of problems and limitations, a score can describe a collective and thus make it comparable to some extent. Because of the different compositions of the collectives, the value of scores for quality control is limited. The more physiological variables a score contains the more difficult it is to distinguish between the effect of therapy and the clinical course. Therefore, repeated measurement of scores is not useful to verify a therapeutic effect, but it can indicate a trend regarding the endpoint. PMID- 8717178 TI - [Mechanism of action of sennosides]. AB - A review of the recent progress in the study of the mode of action of the sennosides, the active constituents of the senna drug, is presented. An interaction between rhein anthrone, the active metabolite of the sennosides, and the immune cells of the colon is suggested as a base for laxative activity. PMID- 8717179 TI - [Listeriosis 1985-1995: microbiologic and epidemiologic aspects]. AB - Listeriosis is an emerging foodborne infection caused by L. monocytogenes (L. m.), mainly identified in industrialized countries. It is a severe disease (meningitis, septicaemia, abortion) which preferentially affects individuals whose immune system is perturbed (pregnant women, newborns, immunocompromized patients and the elderly). Epidemiology is characterized by a background of sporadic cases on which may be surimposed outbreaks. Progresses in microbiology during the last decade (detection and typing of L. m., better understanding of L.m. ecology...) and epidemiological investigations (increased use of case control studies) demonstrated that all kind of foods, at each step of the food chain, can transmit the disease. In many respects, L.m. differs from most recognized foodborne pathogens: it is ubiquitously present in nature, resistant to various kind of environments, microaerophilic and psychrophilic. Its tenacity in industrial environment and its capability to survive in food over extended period of time under adverse conditions made this bacterium the hottest topic for industrials during the last decade. PMID- 8717180 TI - [Total and prolonged parenteral nutrition at the hospital and at the patient's home: its socio-economic importance]. AB - Total parenteral nutrition with "All in one" nutritive mixtures is used in France from 1970. These advances ensure a more simple treatment for intensive care units. Nutritive mixtures provide the caloric and nitrogen daily requirements added with micronutrients (electrolytes, trace elements and vitamins) and this technique allows home parenteral nutrition. "All in one" nutritive mixtures have reduced the infectious complications and the industrial quality control during parenteral solutes admixing has decreased again this infectious risk. So, nutritive mixtures result in sparing of nurse time, material for preparation and antibiotic costs. Moreover, although the costs of home parenteral nutrition are considerable, this technique still reduces annual costs by 50% to 70%, avoids prolonged hospitalization and allows the patients to resume a more normal existence at home. Development of new technologies and nutritive mixtures for specific malnutrition are now required to improve treatment of these patients with insufficiency digestive tract. PMID- 8717181 TI - [Eulogy of Andre Soulairac (1913-1994)]. PMID- 8717182 TI - [Treatment of genital prolapse. Long term results]. AB - From 1975 to 1984 case histories of 237 women operated on for major genital prolapse were looked over. 142 formed the subject of prolonged observation of 4 years and more. 63 over ten years. Prevention of static disorders and tissue atrophy has shown itself to be very useful. The procedure for doing this surgical treatment has been dictated by the clinical, urodynamic and per-operative findings. Vaginal surgery treatment was selected for minor uterine prolapse and for very old patients. Most of them had an operation either through abdominal duct or, more often, mixed abdomino perineal duct. The uterine cervix replacement was realized through presacral cervicopexy, by means of non resolving prothesis. In that manner, an anatomical repair can be achieved. We report the excellent long dated results of this method. Posterior perineorraphy remains a high grad process. His result depends upon the quality of perineal muscles. More difficult is the vesical ptosis and urinary stress incontinence treatment, carried, at one and the same time, through intervesico-vaginal prothesis and/or retropublic pexy. Long dated failures depend usually more on the sphincteral insufficiency than on the recurrence of the cervico-vesical ptosis. Perineal and vesico-sphincteral re education on one hand, substitutive hormonotherapy on the other hand, add a "plus" essential to the surgical repair. Sphincteral bladder insufficiency remains and ill controlled handicap. These desorders are more the consequence of constitutional fragility of connective tissue rather than obstetrical traumatism. PMID- 8717183 TI - [Platelet factor 4, reversible inhibitor of megakaryocytogenesis, protector of megakaryocytes during chemotherapy]. AB - Development of megakaryocyte (MK) from CD34+ cord blood (CB) cells in both plasma clot culture and liquid culture was significantly inhibited by human platelet factor 4 (PF4) and human transforming growth factor beta 1 (TGF beta 1). Inhibition of cell growth by PF4 was reversible judging from the fact that the CD34+ cells preincubated with PF4 could regenerate colonies after washing and replating into the cultures. By contrast, TGF beta 1-pretreated CD34+ cells gave rise to few colonies following replating. Moreover, incubation of CD34+ cells with PF4 in liquid culture caused an increase in the number of both stem cell factor (SCF)-binding cells and CD34 antigen-bearing cells, and exhibited greater capacity to form MK colonies than control after the treatment of 5-FU. In vivo in mice, twice injections of PF4 at 40 micrograms/kg resulted in a significant increase in the number of colony-forming cells with high proliferative potential (HPP-CFC) and colony-forming unit-megakaryocyte (CFU-MK) in bone marrow. In exponentially growing human erythroleukemia cells (HEL), the addition of PF4 prolonged cell cycle progression and therefore resulted in an increased cell population in S phase, as determined by flow cytometric analysis. Different from PF4, TGF beta 1 blocked more cells in G1 phase. These results demonstrate that PF4 and TGF beta 1 inhibit MK development from CD34+ CB cells by different mechanisms and suggest that PF4, unlike TGF beta 1, exerts its inhibitory effect on cell growth in a reversible and S phase-specific manner by which it protects stem cells and MK progenitor cells from 5-FU cytotoxicity. PMID- 8717184 TI - [Recent data on the physiopathology of septic shock]. AB - This review is going on to emphasize recent advances of the pathophysiology of septic shock (SS) which goes-between the systemic inflammatory response syndrome (SIRS) and the multisystem organ failure (MOF). By several recent studies, our knowledge of the cellular and molecular pathophysiology of the SS has dramatically increased. Bacterial products, immunocompetent cells, soluble mediators, and cell-cell interactions between blood cells and endothelium have been reviewed. The metabolic disorders following SS can also be observed during SIRS. Still further, these abnormalities do not accurately predict prognosis, except some of them. A wealth of pre-clinical data suggests the efficacy and potentially useful therapeutic strategies like specific immunotherapy (anti endotoxin, antimediators). Discrepancies between animals models and unexpected and disappointing clinical trial results during SS are discussed. Blocking simultaneously the hole metabolic patterns of SS which is a complex, multisystem, and multifactorial pathologic process seems to be a utopian situation. Furthermore SS occuring more frequently as a complication of nosocomial infection, the high cost of such a novel therapeutic strategy has to be taken into account. PMID- 8717185 TI - [A new concept: treatment using differentiation of the malignant cell in man]. AB - Against the dogma of the irreversibility of malignant cells, although genetical abnormalities, human leukemic cells have been differenciated. According to Leo Sachs proposals on the proliferation-differentiation balance impairment in malignancies we have shown various orientation for the differentiation therapy: inhibition of malignant cells using low dose of Cytosine Arabinoside in acute myeloid leukemia; suppression of the growth factor effect such as the role of interferon in hairy cell leukemia; targetted differentiation process by all trans retinoic acid in acute promyelocytic leukemia with the obtention of complete remission in all cases; differentiation of malignant and normal cells by G-CSF in acute myeloid leukemia in elderly patients. Differentiation therapy induces a modulation in the cell program. Cells mature, becoming differentiated, die, disappear and are replaced by normal cells. The addition of differentiation treatment to chemotherapy greatly improves the chance of long term survival. PMID- 8717186 TI - [Results, evolution of assisted reproduction techniques]. AB - During the last decade IVF and other assisted conception treatment (ACT) methods have become an internationally accepted form of treatment of infertile couple. The final definition of success following ACT is the couple who having received treatment have a live child. As IVF treatment gradually becomes more successful and as the number of babies born as a result of this treatment increases more attention is being focused on the outcome of ACT pregnancies. Recent and futur development of ACT are related. PMID- 8717187 TI - [Capabilities and limitations of research]. AB - Assisted human reproduction (AHR) currently solves numerous difficulties caused by sterility in couples. However, it poses some problems whose solutions involve three main directions in research. Fundamental research explores spontaneously occuring phenomena. It shows that over and above common general points of sexual reproduction, mechanisms can differ from one to another. Technical research perfects artificial empirical methods which lead to fertilization, implantation and pregnancy. It shows that success varies from one species to another. Concerning some technologies such as intracytoplasmic sperm injection, successful results began with humans where they seem to be easier to obtain. Research of risks cheks, in animals, the absence of adverse effects of the above techniques for conceptus. It has confirmed the harmfulness of some environmental factors for gametes and zygotes. Moreover, it shows that these negative effects vary between species and within them for strains and sex. For instance, the cryopreservation of mouse embryos leads to changes in the offspring concerning morphophysiological and behavioral features, some of them appearing in elderly subjects. Indeed, these changes vary as a function of strain and sex. These results as a whole show that experimentation on animals can indicate research areas and also give rise to the need for caution. However, if we want to act on human reproduction, they also show, because of the variations in reactivity from one species to another, that the animal model is insufficient and that research in man is essential. From this point of view, a long term follow up of AHR children seems necessary, as well as the need for reflexion concerning the possibility and conditions of research on human embryos. PMID- 8717188 TI - [Maternal risks of medical assistance with procreation]. AB - It is now well demonstrated that assisted reproductive technologies, namely intra uterine insemination, in vitro fertilization and embryo transfer, and gamete intra fallopian transfer (GIFT) are the source of maternal complications, sometimes life-threatened. All documented hazards concern the short term. Few are related to the techniques: local, regional or general infections, severe hemorrhages. Most are the consequence of controlled ovarian hyperstimulation: ovarian hyperstimulation syndrome (OHSS), thrombosis, multiple pregnancies. High range multiple pregnancies could be prevented by a reduction in the number of embryo transfered, which is now currently limited to two or three in most centers. Similarly, good results can be obtained with intrauterine insemination, while maintaining the multiple pregnancy rate at the physiologic level, about 1%, by avoiding poly-ovulation. In comparison with spontaneous pregnancies, ART pregnancies are more complicated (toxemia, intra-uterine growth retardation, prematurity). The caesarean section rate is over 25% in singleton deliveries, 40% for twins and 89% for triplets. With a long term follow-up of 15 years, no hazard has been demonstrated. Concerns about ovarian epithelial cancer, breast cancer, and premature menopause remain hypothetic. Further studies are in development in this field. PMID- 8717189 TI - [Questions about the future of the child]. AB - Among all the new artificial reproductive techniques the intracytoplasmic sperm injection, ICSI, raises the most questions. Two kinds of potential hazards have to be discuted: 1/ the risks depending on the indications; a lot of male sterilities are of genetic cause, chromosomal abnormalities, cystic fibrosis, ciliary dyskinesia. In such cases the transmissibility is not limited to a risk of sterility. The infertility is part of a somatic syndrome. 2/ the technical risks; invasive procedure, intraoocyte drug injection, eventually use of immature gamete (spermatid). The first kind of potential hazards are predictable in matter of nature, much less in matter of incidence. The second kind of risk are yet unpredictable. The obligatory risk assessment of this new technique needs a long term surveillance plan. PMID- 8717190 TI - [Assisted reproduction vigilance]. AB - Reports on assisted reproduction outcome show that some complications are more frequent than in the general population. These complications include preterm birth, low birthweight and perinatal mortality. Concern has been expressed that exposure to fertility drugs might be associated with the risk of ovarian cancer. Therefore it is important that new assisted reproduction technics (A.R.T.) be evaluated as rigorously and completely as every new drug before it is on the market. Evaluation of short term efficacy and safety of new A.R.Ts should be done using the methodology proposed for the evaluation of new drugs. To evaluate long term safety it is recommended that every A.R.T. center keeps on a standardized form informations on every attempt and its outcome. This information could be the basis for case control surveys looking at the long term consequences of A.R.T. PMID- 8717191 TI - [Ethical reflections]. AB - In the practice of medically assisted conception it is important to avoid harming both the mother and the future child. A rigorous and permanent evaluation of the techniques and their consequences afford the basis of a detailed and complete information of the couples before obtaining written consent. Medical records must include the state of the newborn. Records can be consulted later on according to confidentiality, administrative rules and avoiding any action which can determine anxiety. PMID- 8717192 TI - [Conclusions of the meeting dedicated to problems posed by Assisted Reproduction]. PMID- 8717193 TI - [Driving of a motor vehicle after implantation of a cardioverter-defibrillator in malignant heart rhythm disorders. Criteria for the medical assessment of driving fitness in Europe]. AB - AIM OF STUDY: To investigate how medical permission to drive in patients with a cardioverter defibrillator is handled in various European countries. METHODS: A specially developed questionnaire was sent to all 46 delegates of the pacemaker groups of the European Society of Cardiology. They were asked to provide information on the procedures and criteria used in the various countries regarding a driving ban after cardioverter defibrillator implantation (ICD). RESULTS: 39 answers (83%) were received from representatives of 24 countries. 22 (56%) of those replying advised their patients not to drive. A permanent driving ban was recommended by 13 (33%), a temporary one of 3-18 (mean 9 +/- 4) months by 26 (67%). Presyncope was named as a criterion for a ban by 15 (38%), syncope by 13 (33%) and multiple defibrillator shocks by two (5%). Contrary to medical advice not to drive about one third of patients resumed driving, half of them after 6 months. 12 months after implantation most of the patients were again driving. Defibrillator discharges occurred in 2 patients, without consequence. One patient had a fatal car accident, unrelated to loss of consciousness or defibrillator shock. CONCLUSIONS: 1. ICD discharges are an extremely rare cause of driving accidents. 2. About half of the cardiologists recommended a driving ban for a mean of 9 months after ICD. 3. Despite medical ban about half of the patients resumed driving after 6 months. 4. There are as yet no uniform criteria in Europe for judging fitness to drive. PMID- 8717194 TI - [Thrombolysis of an extensive venous thrombosis of the lower body in an anomaly of the vena cava inferior]. AB - HISTORY AND CLINICAL FINDINGS: A 41-year-old man was admitted to hospital for acute pain in the right flank. At first urolithiasis was suspected but excluded by sonography and excretion urography. Computed tomography (CT) was performed because of increasing pain and swelling of the left leg. It showed bilateral thrombosis of the deep leg and pelvic veins as well as of the infrarenal inferior vena cava (IVC) and the right renal vein. The patient's general condition was impaired. The circumference of the left leg was greater than that of the right by 5 cm 15 cm above the knee joint space and both legs had marked varicosities with congestive dermatitis. INVESTIGATIONS: The concentration of the thrombin antithrombin complex was 13.4 micrograms/l, D-dimer 124 micrograms/l, prothrombin fragment F1+2 3,5 nmol/l and fibrin monomere 27.7 micrograms/ml, as expression of a manifest thrombosis. CT with contrast medium demonstrated the previously sonographically shown extent of the thrombosis. In addition the hepatic IVC segment was absent. The azygos vein was enlarged and there was an extensive collateral circulation. TREATMENT AND COURSE: Systemic thrombolysis (streptokinase for 3 days, then urokinase) for 11 days dissolved the thrombi and CT now demonstrated complete recanalization of the caudal deep vein system and the IVC abnormality (absence of hepatic segment). After intravenous thrombolysis oral anticoagulation with phenprocoumon was started and will be continued all the patient's life. PMID- 8717195 TI - [Trimethoprim-sulfamethoxazole-induced cholestatic hepatitis. Clinico immunological demonstration of its allergic origin]. AB - HISTORY AND CLINICAL FINDINGS: A 22-year-old woman was given trimethoprim plus sulphamethoxazole for a urinary infection (160 and 800 mg, respectively, daily), drugs she had not previously taken. After 2 weeks she noticed a rash of small spots on her trunk. In addition she had nausea and vomiting. The rash faded within 2 days of stopping the drug, but progressive jaundice developed. INVESTIGATIONS: SGPT and SGOT concentrations rose to maximally 328 and 83 U/l, total bilirubin to maximally 5.9 mg/dl. There was no evidence for viral hepatitis (B or C, cytomegalovirus, Epstein-Barr), autoimmune hepatitis or primary biliary hepatitis. Liver biopsy showed central acinar cholestasis, which suggested drug induced liver damage. COURSE: The patient's symptoms regressed over several weeks without any specific treatment and 8 weeks after onset of the rash the laboratory tests also became normal. The allergic cause of the cholestatic hepatitis was confirmed by a lymphocyte transformation test. CONCLUSION: Clinical suspicion of drug allergy as cause of a cholestatic hepatitis can be confirmed reliably and without any risk to the patient with the lymphocyte transformation test. PMID- 8717196 TI - [Diagnosis of lung abscesses]. PMID- 8717197 TI - [Therapy of lung abscesses]. PMID- 8717198 TI - [Adenosine as an anti-arrhythmia agent]. PMID- 8717200 TI - [Revocation of a work disability certificate and the medical duty of confidentiality. The judgement of the Oldenburg Regional Court of 24 January 1995]. PMID- 8717199 TI - [The sympathetic-adrenergic system in heart failure]. PMID- 8717201 TI - [Isolated elevated diastolic blood pressure]. PMID- 8717202 TI - [Follow-up phlebography after the conservative treatment of a deep venous thrombosis of the leg?]. PMID- 8717203 TI - [Recurrent immunothrombocytopenia]. PMID- 8717204 TI - [Lipoprotein(a) and hemostasis-activation markers in angina pectoris]. PMID- 8717205 TI - [Analysis of protein primary structures by mass spectrometry]. PMID- 8717206 TI - [Biochemistry of neurotrophins]. PMID- 8717207 TI - [Adenylyl cyclase associated protein (CAP) and their homologs]. PMID- 8717208 TI - [Possible mechanism of protein transport to nucleolus]. PMID- 8717209 TI - [Preparation of F(ab')2 mu fragments of IgM monoclonal antibodies]. PMID- 8717210 TI - [Role of nitric oxide (no) in urology]. PMID- 8717211 TI - [Early detection of prostate cancer-results of a prostate specific antigen-based detection program at a "multiphasic screening"]. AB - Determination was made of serum PSA in the early detection of prostate cancer in a population of 1,227 men aged 55 years or older who visited a "human dock". "Human dock" is a unique health examination facility open to public in Japan. Recommendation for biopsy was made based solely on this parameter when its value exceeded 2.0 ng/ml in IMx immunoenzymetric assay (Dinabot Co.). Of these, 162 (13.2%) had elevated values. The proportion of males with serum PSA greater than 4.0 ng/ml was only 3.6%. Of the 109 males who underwent ultrasound guided biopsies, 17 cancers were detected, the cancer detection rate thus being 1.4%. Most of these cases (82.4%) were clinically localized cancers. Eight patients with cancer had serum PSA levels below 4.0 ng/ml including 6 less than 3.0 ng/ml. Radical prostatectomy was conducted on 14 patients. All had histologic features of clinically significant cancer and 64.3% were pathologically confined. Oriental elderly males appear to have lower serum PSA than western counterparts. The optimal cutoff of serum PSA for early detection should be examined further in oriental male populations. Differences in the incidence of prostate cancer between ethnic groups may have been overestimated in previous studies owing in part to unawareness by the physician and general public. Though the application of PSA for early detection will likely disclose greater numbers of prostate cancers in Japan, whether early detection reduces mortality by minimizing risk of death from cancer remains a point to be clarified. PMID- 8717212 TI - [Preoperative use of erythropoietin (rH-EPO) and determination of optimal doses in patients with benign prostatic hypertrophy for future autologous blood transfusion]. AB - BACKGROUND: Recently, autologous blood transfusion has been widely endorsed, because of the adverse effects attributed to homologous blood transfusion. We found that the administration of recombinant human erythropoietin (rH-EPO) permitted the preoperative collection of an adequate volume of autologous blood in a short period of time. This reduced or eliminated the need for homologous blood transfusion. METHODS: To determine optimal dosage of rH-EPO, I conducted a randomized, controlled trial in 86 patients with benign prostatic hypertrophy (BPH) scheduled for transurethral resection of the prostate (TUR-P). The patients from whom approximately 10% of their total blood volume was removed, received six different doses of rH-EPO either intravenously or subctaneously. All patients received iron sulfate 100 mg orally once a day during the study. RESULTS: I found that 9,000 IU of rH-EPO given intravenously daily or 10,500 IU of rH-EPO injected subcutaneously every third day for a week were optimal dosages. These schedules ennabled us to withdraw approximately 400 ml of blood prior to operation without adverse effects. CONCLUSION: I conclude that the efficacy of rH-EPO is greater when administered subcutaneously rather than intravenously. It is clear that rH EPO increases the ability of patients about to undergo selective surgery to donate greater amounts of blood for future autologous transfusion. PMID- 8717213 TI - [Evaluation of reliability of IPSS by 24-hour uroflowmetry]. AB - BACKGROUND: Although the international prostate symptom score (IPSS) is now often used to assess the symptoms of BPH, whether or not patients answer the questions correctly has not been validated objectively. METHODS: Reliability of IPSS was evaluated by 24-hour uroflowmetry in 20 hospitalized male patients. Six of them had prostatic hypertrophy or cancer, and the evaluation was performed before and after TURP or hormonal therapy in these six patients. The objective scores for frequency and nocturia were obtained from the time recorded on IC card. The objective score for intermittency was calculated from the uroflow curves. RESULTS: The answer about frequency was not correct compared with the objective scores for frequency. The answers about nocturia and intermittency were almost the same as the corresponding objective scores. The answer for weak stream correlated with the average of peak flow rate. However, the threshold of peak flow rate for "weak stream" fluctuated markedly before and after the treatment in the same individuals. The answer for hesitancy had no correlation with the hesitation time. Patients seemed to understand the question translated in Japanese as "have you need force to urinate?" CONCLUSION: Before wider use of IPSS in Japan, the correct translation of the questions and verification of the usefulness of the questions in large number of Japanese patients seem necessary. PMID- 8717214 TI - [Pelvioureteral function after ureterotomy--experimental study simulating endopyeloureterotomy]. AB - BACKGROUND: Endopyeloureterotomy has been established as a valuable procedure for ureteropelvic junction or upper ureteral stenosis. In order to evaluate ureteropelvic function after endopyeloureterotomy, I examined electromyographic change in ureteral peristalsis, measured urine bolue volume, and monitored ureteral compliance in dogs following ureterotomy. METHODS: Twelve adult mongrel dogs were anesthetized with pentobarbital, and the upper part of the left ureters was incised longitudinally for a distance of 2 cm. After the incision of the left ureter, a polyurethane splint was inserted through the kidney to the lower part of the ureter and left for four weeks. At 5 (n = 4), 12 (n = 4), and 24 (n = 4) weeks after the operation, electromyography in ureteral peristalsis, urine bolue volume, and ureteral compliance at the incised part of the ureter were investigated. During the measurement of peristaltic frequency and urine bolus volume, saline was infused to the renal pelvis at a constant rate through a nephrostomy. I also investigated these factors in 4 dogs without ureterotomy and defined this group as control. RESULTS: When the infusion rate increased, ureteral peristaltic frequency and/or urine bolus volume gradually increased in the groups of 12, 24 weeks and control group after the operation; in the group of 5 weeks, ureteral peristaltic frequency and bolus volume had not increased significantly. Peristaltic velocity at the incised area decreased significantly at 5 weeks and 12 weeks following the operation compared to the velocity in the control group, but recovered at 24 weeks. Ureteral compliance decreased significantly at 5 weeks compared to that in the incised area of control dog ureter, but increased gradually and recovered at 12 weeks. In the incised ureter the defect was healed with granulation tissue within 5 weeks, and covered by smooth muscle within 12 weeks. CONCLUSION: These results indicate that ureterotomy had harmful effects on ureteral peristalsis 5 weeks following surgery, but that ureteral function generally returned at 12 weeks, and recovered completely at 24 weeks. It was supposed that endopyeloureterotomy would improve ureteral function in patients with ureteropelvic junction and upper ureteral stenosis. PMID- 8717215 TI - [Prognostic factor in patients with prostate cancer]. AB - BACKGROUND: Histological grading, especially the Gleason Score (GS), is considered to be of value in determining the prognosis of patients with prostate cancer. However, subjective histological grading is characterized by low reproducibility. On the other hand, estimate of Mean Nuclear Volume (MNV), which was developed by Gunderson and Jensen based on a sterological technique, is a very easy method with high reproducibility. Furthermore, it had been reported that MNV provides accurate prognosis of bladder cancer. In this study we compared MNV with two histological grading methods in determining the survival of prostate cancer. METHOD: A retrospective, prognostic study of 49 patients with prostate cancer diagnosed by Transurethral Resection of the Prostate (TUR-P) or Needle punch biopsy between Jan. 1983 and Jul. 1994 was performed. Unbiased estimates of MNV were compared with age at the time of diagnosis, clinical stage, histological grading according to, the general rules for clinical and pathological studies on prostatic cancer of the Japanese Urological Association (J.U.A.) and GS on the prognostic value. RESULTS: Although age at the time of diagnosis, J.U.A. classification and GS had no value as prognostic criteria, clinical stage (p = 0.0018) and MNV (p = 0.0087) correlated significantly with survival of prostate cancer. CONCLUSION: Results of this study indicate that estimates of MNV and clinical stage are prognostically superior to age at the time of diagnosis and morphologic grading of malignancy, such as J.U.A. classification and GS, in prostate cancer. PMID- 8717216 TI - [Murine "sympathetic orchitis" induced by unilateral testicular injury and autoimmune response]. AB - BACKGROUND: Experimental autoimmune orchitis (EAO) has been studied as an animal model for human male immunological infertility. Most EAO models have been induced by immunization with testis antigens in artificial adjuvants. In this paper, we report a more clinical EAO model. METHODS: Ten to 20 needle punctures were made to the unilateral testis of mice and it was crushed by a needle-holder. RESULTS: Contralateral EAO (so-called "sympathetic orchitis") was gradually induced starting on Day 28. Delayed type hypersensitivity (DTH), one of the cell-mediated immunities, to autologous testicular cells (TC) as well as anti-TC antibodies, humoral immunity, were both detected in those mice. Repeated crush(es) of the ipsilateral testis two weeks later (and four weeks later) as a booster did not enhance the contralateral lesion or autoimmune responses. CONCLUSION: Our present injury model mimics clinical testicular trauma; therefore, this testicular injury model can be very useful in studying the immunological mechanism of EAO and of human immunological male infertility. PMID- 8717217 TI - [Sclerotherapy with 3% polidocanol for hydrocele testis]. AB - BACKGROUND: We studied the clinical efficacy of sclerotherapy with injection of 3% polidocanol for hydrocele testis. METHODS: From July, 1992 to March, 1995 sclerotherapy with single injection of polidocanol was performed for 11 patients with 12 hydrocele testis on an outpatient basis. We instilled 3 or 5ml of 3% polidocanol after complete removal of fluid in the hydrocele testis. RESULTS: Complete disappearance on ultrasonography was observed in 75% of the hydrocele testis 6 months after this sclerotherapy. There was neither pain during instillation of 3% polidocanol nor any other complication. Two patients with fluid reaccumulation underwent hydrocelectomy 16 and 6 months after sclerotherapy, respectively. CONCLUSION: This procedure seems to be a safe and useful technique as primary treatment for hydrocele testis. PMID- 8717218 TI - [The experimental evaluation of the difference in closing dynamics of internal urethral ostium on the urinary continence and on the ejaculation induced by an administration of phenylephrine]. AB - BACKGROUND: The internal urethral ostium is usually closed both on the ejaculation and on the urine collection, although the difference of the closing characteristics have not been known. METHODS: The difference in closing style of bladder neck (internal urethral ostium) between on the phenylephrine administration corresponding state to the ejaculation and on the filling of urinary bladder corresponding state to the urinary continence was evaluated by Stereo-UPP method using mongrel dogs. RESULTS: On phenylephrine administration, the length between verumontanum and bladder neck was lengthened, and a remarkable pressure increase was observed in the area. On the urinary continence filling physiological saline in the bladder, the length of the urethra on the UPP curve of the anterior wall of urethra was lengthened (10%) and the base of bladder was gradually become flat. The pressure in the bladder neck was not relatively increased. CONCLUSIONS: The internal urethral ostium was closed both on the phenylephrine administration and on the urine collection, although the closing characteristics of each was very different. It was demonstrated that on the phenylephrine administration the area between the verumontanum and the bladder neck was lengthened, together with the rise of pressure in the area, whereas on the urine collection bladder neck was closed in low pressure by flattening the base of bladder. PMID- 8717219 TI - [Clinical experience with the artificial urinary sphincter AMS 800 for male incontinence]. AB - BACKGROUND: The aim of this study is to evaluate the therapeutic efficacy of artificial urinary sphincter AMS 800 implants for male patients with urinary incontinence. METHODS: Eleven male patients with urinary incontinence were treated by implantation of the artificial urinary sphincter AMS 800 between 1988 and 1992. Patient age at the surgery ranged from 14 to 79 years, with a mean age of 58 years. At presentation, 9 patients (82%) had true incontinence and 2 had overflow incontinence, and medical treatments and/or surgical procedures (Teflon injections in 2 and Sling procedure in 1) had been attempted previously elsewhere in all patients. The etiologies of incontinence were post-prostatectomy (transurethral resection in 4 and radical retropubic prostatectomy in 4) in 8 patients (73%) and myelomeningocele, spinal cord injury, pelvic trauma in one each. There were 5 patients with abnormal cystometrogram, and 2 of them were performing intermittent self catheterization. Vesicoureteral reflux was determined in 2 patients preoperatively, which were surgically corrected one year before AMS800 implant in a patient and simultaneously in another patient. The cuff was placed around the bulbous urethra (9), pendulous urethra (1) or bladder neck (1). The device was activated 6 weeks post implantation, and the assessment of therapeutic effects was started 2 weeks after activation. RESULTS: Follow up ranged from 3 weeks to 75 months, with a mean of 56 months, if 3 cases done explantation due to periprosthetic infections were excluded. After AMS 800 implantation 5 patients (45%) were completely continent, 4(36%) required the use of not more than 1 pad per day, while a patient was not satisfied with the results and another patient was not definitive because of early explantation before device activation. Hence complete or near complete continence was achieved in 9 patients (81%). A patient is performing intermittent self catheterization in conjunction with the AMS 800 without any complications up until now. There were 3 periprosthetic infections (27%) associated with 2 cuff erosions, that consequently required explantation. Besides periprosthetic infection, neither complications nor mechanical device failures was experienced. CONCLUSION: In conclusion, the AMS 800 artificial urinary sphincter was safe with reasonable mechanical reliability and offered acceptable and satisfactory urinary control to the selected patients with incontinence. PMID- 8717220 TI - [Clinical significance of PSA-density in differential diagnosis between BPH and early stages prostate cancer]. AB - OBJECT: In differential diagnosis of BPH and early stages prostate cancer (PC), PSA-density (PSAD) was evaluated in 63 cases with BPH and 82 cases of PC (stage A: 8, B: 17, C: 17, D: 40). METHODS: Serum PSA values were determined by MARKIT-F PA, and prostate volume was calculated by transabdominal ultrasonography, in which every glands including peripheral zone was visualized in the transverse and sagittal planes, and predicted prostate volume was calculated by three dimension (a x b x c x 0.52). PSAD was determined by serum PSA divided by prostate volume. RESULTS: PSAD values were 0.106 +/- 0.006 (mean +/- SD) in BPH, 0.538 +/- 0.094 in stage A and B of PC, and 2.973 +/- 0.764 in all PC cases. There was a statistical significance (p < 0.005) between BPH and each other groups of PC by student's t-test. In using 0.208 (mean +/- 2 SD of PSAD in BPH group) as a cut off value, the detection sensitivity was 84% in stage A and B of PC and the specificity was 97% using BPH groups as a control, therefore the efficacy was 93%. In 18 out of 19 cases with BPH having PSA values more than 3.6 ng/ml (false positive group), PSAD values were less than 0.208. CONCLUSION: PSAD is suggested to be a useful tool for differential diagnosis of BPH and early stages of PC. PMID- 8717221 TI - [Primary retroperitoneal pure choriocarcinoma]. AB - A thirty-year-old clerk presented with a history of left supraclavicular lymph node swelling of a month duration and newly detected multiple lung and mediastinal tumors. Abdominal computed tomography showed two retroperitoneal masses, but no abnormality in both testes was found by physical and ultrasonographic examination. Biopsy of supraclavicular lymph nodes revealed pure choriocarcinoma. Furthermore, beta-subunit of human chorionic gonadotropin was elevated up to 700 ng/ml but alfafetoprotein was normal, resulting in the diagnosis of primary retroperitoneal pure choriocarcinoma. Two cycles of combination chemotherapy with etoposide (VP-16), ifosfamide and cisplatin (VIP) was given and followed by bleomycin, etoposide and cisplatin (BEP) (2 cycles) because of cystitis symptom due to ifosfamide. Since post chemotherapeutic evaluation revealed only partial response, we surgically removed all residual masses in the lung, mediastinum and retroperitoneum, which histologically proved to be necrotic fibrous tissue. He has remained disease-free 15 months after surgery without additional therapy. In order to improve prognosis of primary retroperitoneal pure choriocarcinoma, intense chemotherapeutic regimen like VIP should be given, regularly repeated and followed by aggressive surgical resection of residual masses. PMID- 8717222 TI - [Spontaneous rupture of ureter caused by ureteral cancer]. AB - A 63-year-old man was admitted with right flank pain, nausea and chill. CT scan revealed right hydronephrosis and rupture of ureter, but tumor or stone was not detected in the CT scan. However retrogradepylelography revealed right lower ureteral tumor, and this patient was treated by right nephroureterectomy and partial cystectomy. Histopathological examination of ureteral tumor showed transitional cell carcinoma. Spontaneous rupture of ureter due to ureteral cancer is a rare case, which has not been reported in the Japanese literature. Clinical study was performed about cases of spontaneous rupture of ureter which have been previously reported. PMID- 8717223 TI - [Immunotoxicity of chemicals]. AB - Immunotoxicity is the detrimental effects of xenobiotics on immune functions of hosts, which result in enhanced susceptibility to infectious agents and tumours. A variety of environmental chemicals, such as pharmaceuticals, food additives, contaminating chemicals in food and drinking water, pesticides, and airborne chemicals, have been reported to exert immunotoxic effects on experimental animals and humans. In this paper, the mechanisms of immune responses, the effects of typical immunotoxic chemicals on the immune systems, and the methods for detecting immunotoxicity were described. In addition, the strategies and guidelines for immunotoxicity assessment were briefly outlined and discussed. PMID- 8717225 TI - [Teratogenicity study of stevioside in rats]. AB - Teratogenicity of stevioside was examined in rats. Stevioside dissolved in distilled water was given to pregnant Wistar rats by gavage once a day from day 6 through 15 of pregnancy at doses of 0, 250, 500 and 1000 mg/kg/day. The pregnant rats were sacrificed on day 20 of pregnancy and their fetuses were examined for malformation. Stevioside caused no increased incidences of fetal malformation, and no toxic signs in the pregnant rats and the fetuses. It was concluded that stevioside has no teratogenicity in rats when given by gavage. The no observable adverse effect level was estimated to be over 1000 mg/kg/day for both pregnant rats and rat fetuses. PMID- 8717224 TI - [Twenty-eight days repeated dose toxicity test of N (fluorodichloromethylthio)phthalimide in rats]. AB - N-(Fluorodichloromethylthio)phthalimide (Fluor-folpet) has been widely used as an anti-mold and anti-bacterial agent. In this study, 28 days repeated-dose oral toxicity study of fluor-folpet was carried out in Slc:Wistar rats. An oral toxicity study for fluor-folpet, the twenty-eight days test, repeated-dose, oral administration, was performed as follows: Five week-old rats, male and female, 10 rats, each/group, were treated with intragastric administration of fluor-folpet with a dose of 0 (1% Sodium CMC, control), 20, 80 and 320 mg/kg, body weight. Recovery test, for 14 days after the last treatment, was examined for the control and the 320 mg/kg groups. The 320 mg/kg groups, both males and females, showed significantly reduced their body-weight gain compared with the control group. In the 320 mg/kg group, five out of 20 male rats and four out of 20 female rats died from dyspnea during the treatment period. In the female rats in the 320 mg/kg group, serum ChE level was decreased to 50% of control level and gamma-GT was increased in a dose-dependent manner, but these serum levels recovered after 14 days non-treatment period. No histopathological change, relating to the treatment, in liver was observed. Increased weight of the kidney and vacuolation in renal tubules were found in both sexes of 320 mg/kg group. Hyperkeratosis and hyperplasia of the stomach epithelium were observed at the dose more than 80 mg/kg in male, and more than 20 mg/kg in female. A supplemental study, repeated dose, oral administration in rats carried out to examine the dyspnea revealed that severe acute toxic damages in epithelium of nasal cavity and meatus nasopharyngeus were induced by intragastric administration of fluor-folpet. Fluor folpet is shown to be cytotoxic. In conclusion, the no-observed-effect level (NOEL) for fluor-folpet was not found under the experimental conditions employed in this repeated-dose toxicity study. PMID- 8717226 TI - [Hepatic lesions in F344 rats treated orally with beta-cyclodextrin for 13 weeks]. AB - A 13-week oral toxicity study of beta-cyclodextrin was carried out in F344 rats at the dose levels of 10, 5, 2.5, 1.25, 0.6 and 0% in powdered diet. Each group consisted of 10 males and 10 females. All animals survived at the end of the experiment, while a slight decrease in body-weight gain was observed in males of the 10%- and 5%-groups. Dose-dependent increases in serum levels of GOT, GPT and alkaline phosphatase were observed in treated animals of both sexes, and increases in serum levels of urea nitrogen and relative liver weights in treated males. Histopathologically, a dose-dependent increase in the severity of inflammatory cell infiltration was seen in the liver of treated animals, focal hepatocellular necrosis being detected in both sexes of the 10%-group and females of the 5%-group. These findings indicate that beta-cyclodextrin causes hepatocellular injury to rats when it is orally administered. PMID- 8717227 TI - [A 13-week subchronic toxicity study of josamycin in F344 rats]. AB - A 13-week subchronic toxicity study of josamycin was performed in male and female F344 rats to determine the maximum tolerable dose (MTD) for subsequent investigation of the carcinogenicity. As animals refused to take diet containing 5.0% josamycin in our preliminary study, dose levels in the present study were determined as 0, 0.16, 0.32, 0.63, 125 and 2.5% in diet. Rats were randomly allocated to 6 groups, each consisting of 10 males and 10 females. No animal died during the administration period and no group showed significant changes in body weight gain. Definite toxicity of josamycin was not noted in hematological and serum biochemical examinations. Histopathological examinations revealed no particular findings related to josamycin administration except cecal enlargement in the 1.25 and 2.5% groups. based on the results of the present study, it was concluded that the MTD of josamycin in 2.5% in diet, because the dietary dose level of 2.5% proved to exert no significant toxicological signs. PMID- 8717228 TI - [Cell proliferative activities of cholangiofibrosis induced in rats treated with bromodichloromethane]. AB - To clarify whether bromodichloromethane (BDCM)-induced cholangiofibrosis progresses to cholangiocarcinoma, further morphological examinations were performed on the livers obtained from our previous experiment. The livers of Wistar rats fed diet containing 2200, 550, 140 or 0 ppm of microencapsulated BDCM up to 24 months were examined at months 6, 12, 18, and 24. The liver sections were stained with H-E, PAS and Azan, and were subjected to immunostaining using antiproliferating cell nuclear antigen (PCNA) monoclonal antibody for determination of the PCNA labeling index of bile duct epithelia, as well as silver staining for nucleolar organizer regions (AgNORs). At month 6, the severity of hyperplasia of atypical bile duct epithelia in the 2200 ppm group was marked, their PCNA labeling index being highest (68.5). The number of bile ducts gradually decreased, and the severity of fibrosis became more marked, with prolongation of the treatment. The PCNA labeling index in hyperplastic bile ducts in this group also decreased to 31.5 at month 24. The number of AgNORs in the nuclei of bile duct epithelia in the 2200 ppm group was highest at month 6, but decreased thereafter. The present study suggests that the possibility of the progression from cholangiofibrosis to neoplastic lesions is extremelly low. PMID- 8717229 TI - [Toxicity in rats fed diet containing iron lactate for 26 weeks]. AB - In order to characterize the toxicity of iron lactate, a 26-week feeding study was performed in male and female F344 rats. Animals were divided into 2 groups, and given diet containing iron lactate at concentration of 0 or 2%. No animals died during the administration period. Body weight gain was suppressed in both sexes of the 2% group compared with the 0% group. Hematologically, anemia was observed in male of the 2% group. Serum alkaline phosphatase decreased in both sexes of the 2% group. The spleen weight of both sexes and kidney weight of females were higher in the 2% group than in the 0% group. Lipid peroxide increased not only in the liver and the kidney homogenates of treated males and females, but also in the serum of treated females. Histopathologically, iron deposition was observed in the liver, the kidney and the spleen of treated males and females, and in the intestine of treated females. The present results indicate that the iron lactate administration caused iron deposition in the liver and the other several organs, resulting in lipid peroxidation in these organs. PMID- 8717230 TI - [Comparative studies on a single dose toxicity of microsomal Ca(2+)ATPase inhibitor, 2,5-di(tert-butyl)-1,4-hydroquinone and its related analog, mono(tert butyl)-1,4-hydroquinone, in rats]. AB - We performed comparative studies to determine an acute toxicity of microsomal Ca(2+)ATPase inhibitor, 2,5-di(tert-butyl)-1,4-hydroquinone (DTBHQ) and its related analog, mono(tert-butyl)-1,4-hydroquinone (MTBHQ), which are both used as antioxodants. Wistar rats, 5 weeks old, male and female, were used. By a single dose of oral administration, DTBHQ-induced LD50 values (obtained by Lorke method) in male and female rats were estimated 295.1 and 234.4 mg/kg BW, respectively, whereas each LD50 value for MTBHQ was 711.6 and 400.0 mg/kg BW, respectively. MTBHQ-induced deaths occurred from 8 to 20 minutes after administration, however, DTBHQ-induced deaths occurred more delayed from 1 to 5 days after administration. The observed toxic signs of DTBHQ included diarrhea (jelly like), prone position, lacrimation, salivation and abnormal gait (such as reluctance to walk, limping). Localized purpura and loss of the tail (perhaps as a result of necrosis) were also observed. In comparison, MTBHQ elicited prone position, panting, staggering gait and spastic gait. Without loss of the tail montioned above, dead and sacrified rats showed no remarkable changes in macroscopic examination due to exposure to both compounds. PMID- 8717231 TI - [Study on evaluating methods for the quality control of glycoprotein products. (1). Erythropoietin products]. AB - The ability of high performance anion exchange chromatography (HPAEC) with pulsed amperometric detection was studied for evaluation of carbohydrate moieties of erythropoietin (EPO) products. The N-linked oligosaccharides were released from EPO by the treatment with N-glycosidase F. HPAEC analysis of oligosaccharide standards revealed that elution time of sialylated oligosaccharides were dependent on the number of sialic acid, which contributed to the activity of EPO. Using HPAEC, N-linked oligosaccharides of two kinds of recombinant human EPO (rhEPO) produced in chinese hamster ovary (CHO) cells and baby hamster kidney (BHK) cells were compared. The HPAEC profiles of oligosaccharides of these EPO products indicated that there were some differences in the carbohydrate moieties between CHO rh-EPO and BHK rh-EPO. In conclusion, HPAEC method is useful to evaluate the quality of EPO products. PMID- 8717232 TI - [Inspection of carbon monoxide in imported tilapia]. AB - Carbon monoxide in imported tilapia was determined with a gas chromatograph equipped with a molecular sieve 13x column (2.3 m), a methanizer and a FID for the inspection of imported food. The concentration of carbon monoxide in the sample, which was vacuum packed and suspected to be treated with carbon monoxide, was 16 x 10 micrograms/kg in fish meat, and 37 x 10(3) microliters/l in the bubble in the package. On the other hand, carbon monoxide in the reference, which was vacuum packed but was not treated with carbon monoxide, was 10 micrograms/kg in fish meat and 76 microliters/l in the bubble in the package. Carbon monoxide was less than 4 micrograms/kg in two vacuum packed fish meat of tilapia sold in a market in Tokyo. From these results, the suspected sample was concluded to be treated with carbon monoxide for color fixating of protoheme in fish meat. PMID- 8717233 TI - [A study on the establishment of the specification of polyvinylpolypyrroridone as a food additive]. AB - Polyvinylpolypyrroridone (PVPP) has been used in many countries for removing polyphenols in beer and wine, and was permitted as a food additive in Japan in April 1995. Prior to this approval of the compound, we studied procedures for identifying PVPP and its water soluble substances by colorimetry and infrared (IR) spectrometry for the establishment of the specification. PMID- 8717234 TI - [Migration of lead and cadmium from lacquered tableware]. AB - The migration of lead and cadmium from lacquered tableware was investigated. Seven samples of bowls and five samples of chopsticks were purchased on the market. The migration test was carried out with 4% acetic acid at 60 degrees C for 30 min. Lead and cadmium in the test solution was determined by flameless atomic absorption spectrometry. Release of lead was 3.4 and 4.4 ppb from two samples and release of cadmium was 1.3 ppb from one sample. Those samples were chopsticks coated with natural lacquer and colored with pattern. No other samples released lead and cadmium. Evapolation residue was less than 13 ppm in all samples. PMID- 8717235 TI - [A new programme on safety evaluation of pesticides--the IPCS Joint Meeting on Pesticides (JMP)]. AB - International Programme on Chemical Safety (IPCS), an international collaboration in safety evaluation of chemicals, initiated a programme called Joint Meeting on Pesticides or JMP, last autumn. The JMP is an activity contributing to harmonizing evaluation procedures and saving expertise and financial resources, having several unique features. First, the outputs of the scientific evaluations and data covering all major areas related to pesticide safety in one JMP meeting can be applied to the areas of food safety, occupational health, and environmental protection. Second, succinct presentation of the outputs of reliable evaluations in a tabular form, supported by detailed information in the Environmental Health Criteria documents help people in countries, especially those in developing ones, understand the evaluations and rely on them in establishing their guidelines on pesticide safety. This format of the JMP report is a good example for short assessment reviews on chemicals (Concise International Assessment Document) which will be prepared by an international cooperation project based on Agenda 21 decisions. PMID- 8717236 TI - [First drafts of the Environmental Health Criteria (EHC) circulated for comments by IPCS in 1994-1995]. AB - Summaries of first drafts of Environmental Health Criteria (EHC), which were circulated for comments by IPCS in the period of 1994-1995, are presented. EHC drafts on 10 compounds were received in this period. PMID- 8717237 TI - [Estimated production by the official inspection of coal-tar dyes (including dye aluminum lakes) in 1994]. AB - The number of official inspection of coal-tar dyes and their lakes from April in 1994 till March in 1995 were 635 in total. The quantity which passed inspection amounted to 186 ton in Japan. The production of color in each month was summarised in Table 1, and by each producing company in Table 2. The food coal tar dye produced in the largest quantity was Food Yellow No.4, occupying 40.7% in this period. PMID- 8717238 TI - [Ergocalciferol reference standard (Control 941) of the National Institute of Health Sciences]. AB - The raw material for ergocalciferol was tested for preparation of the "Ergocalciferol Reference Standard (Control 941)". Analytical data obtained were as follows: melting point, 117.6 degrees C; UV and infrared spectra, the same as those for JP Cholecalciferol Reference Standard; specific absorbance, E1%1cm = 458 (265 nm); thin-layer chromatography and high-performance liquid chromatography (HPLC), no impurities were detected, respectively; assay, 100.6% by HPLC. Based on the above results, the candidate raw material was authorized as the Japanese Pharmacopoeia Reference Standard (Control 941). PMID- 8717239 TI - [Cholecalciferol reference standard (Control 941) of the National Institute of Health Sciences]. AB - The raw material for cholecalciferol was tested for preparation of the "Cholecalciferol Reference Standard (Control 941)". Analytical data obtained were as follows: melting point, 88.5 degrees C; UV and infrared spectra, the same as those for JP Cholecalciferol Reference Standard (Control 923), respectively; specific absorbance at 265 nm E1%1cm = 471; optical rotation, [alpha]20D = +107.3 degrees; thin-layer chromatography and high-performance liquid chromatography (HPLC), no impurities were detected, respectively; assay, 101.3% by HPLC. Based on the above results, the candidate raw material was authorized as the Japanese Pharmacopoeia Reference Standard (Control 941). PMID- 8717240 TI - [Prednisolone acetate reference standard (Control 941) of the National Institute of Health Sciences]. AB - The raw material for prednisolone acetate was tested for preparation of the "Prednisolone Acetate Reference Standard (Control 941)". Analytical data obtained were as follows: melting point, 237.5 degrees C (decomposition); UV and infrared spectra, the same as those for JP Prednisolone Acetate Reference Standard (Control 903), respectively; specific absorbance at lambda max E1%1cm = 379; optical rotation, [alpha]20D = +115.2 degrees; thin-layer chromatography and high performance liquid chromatography (HPLC), no impurities were detected, respectively; assay, 100.8% by HPLC. Based on the above results, the candidate material was authorized as the Japanese Pharmacopoeia Reference Standard (Control 941). PMID- 8717241 TI - [Betamethasone valerate reference standard (Control 941) of the National Institute of Health Sciences]. AB - The raw material for betamethasone valerate was tested for preparation of the "Betamethasone Valerate Reference Standard (Control 941)". Analytical data obtained were as follows: melting point, 194.3 degrees C (decomposition); UV and infrared spectra, the same as those for JP Betamethasone Valerate Reference Standard (Control 844); optical rotation, [alpha]20D = +79.4 degrees; thin-layer chromatography and high-performance liquid chromatography (HPLC), no impurities were detected; assay, 100.0% by HPLC. Based on the above results, the candidate material was authorized as the JP Betamethasone Valerate Reference Standard (Control 941). PMID- 8717242 TI - [Glycyrrhitinic acid reference standard (Control 941) of the National Institute of Health Sciences]. AB - The raw material of glycyrrhitinic acid was examined for preparation of the "Glycyrrhitinic Acid Reference Standard". The candidate material was evaluated physico-chemically by a collaborative study in which five laboratories participated. Analytical data obtained were as follows: UV spectrum, lambda max = 251 nm and specific absorbance E1%1cm (EtOH) at the lambda max = 145; IR spectrum, specific absorption numbers were at 1719, 1654, 1216, and 1170 cm-1; thin-layer chromatography, some laboratories detected a trace amount of one spot and others not detected; high-performance liquid chromatography, 2-6 impurities were detected and the amount of any impurities were estimated to be less than 0.2% and the total amount less than 0.5%. Based on the above results, the candidate material was authorized as the Glycyrrhitinic Acid Reference Standard of National Institute of Health Sciences. PMID- 8717244 TI - [Ulinastatin reference standard (Control 941) of the National Insutitute of Health Sciences]. AB - The raw material of ulinastatin was examined for preparation of the "Ulinastatin Reference standard". The candidate material was evaluated in collaboration with one domestic laboratory, and the potency of trypsin inhibiting activity was determined to be 3500 unit/vial. Other analytical data obtained were as follows: UV maximum absorption was observed at 276 nm, the molecular weight was estimated to be about 66000 +/- 5000 by gel filtration method. Maximum variance of material contents in 10 vials was 6.52% by means of the weight variation test in JP XII. Based on the above results, this raw material was authorized to be the first "Ulinastatin Reference Standard" of the National Institute of Health Sciences. PMID- 8717243 TI - [Berberine hydrochloride reference standard (Control 941) of the National Institute of Health Sciences]. AB - The raw material of berberine hydrochloride was examined for preparation of the "Berberine Hydrochloride Reference Standard". The candidate material was evaluated physico-chemically by a collaborative study in which five laboratories participated. Analytical data obtained were as follows: UV spectrum, lambda max = 228, 263, 345, and 421 nm and specific absorbance E1%1cm at each lambda max = 814, 794, 722, and 160, respectively; IR spectrum, specific absorption wave numbers at 2845, 1633, 1568, and 1506 cm-1; thin-layer chromatography, some laboratories detected a trace amount of one or two spot and others not detected; high-performance liquid chromatography, 2-5 impurities were detected and the amount of any impurities were estimated to be less than 0.05% and the total amount less than 0.2%. Based on the above results, the candidate material was authorized as the Berberine Hydorochloride Reference Standard of National Institute of Health Sciences. PMID- 8717245 TI - [The human chorionic gonadotrophin reference standard (Control 941) of the National Institute of Health Sciences]. AB - Raw human chorionic gonadotrophin material was examined for preparation of the "Human Chorionic Gonadotrophin Reference Standard (Control 941)". The candidate material was assayed against the 3rd International Standard by the rat ovarian weight method. The potency of the new standard was defined as 1180 international units per ampoule as the result of 18 assays in four collaborative laboratories. PMID- 8717246 TI - [Protamine sulfate reference standard (Control 941) of the National Institute of Health Sciences]. AB - Three kinds of the candidate raw material for protamine sulfate was tested for preparation of the "Protamine Sulfate Reference Standard (Control 941)". The candidates were evaluated by physicochemical tests and anti-heparin tests. Analytical data were summarized in Table 1. Based on the above results, the best one having the highest anti-heparin activity was selected and authorized as the Japanese Pharmacopoeia Reference Standard (Control 941). PMID- 8717247 TI - [Technical implications on the quality standard of new drugs]. AB - New guidelines on the establishment of quality specifications and analytical methods for a new drug application were notified from the Evaluation Division of the Pharmaceutical Affairs Bureau, Ministry of Health and Welfare, on September, 1994. General policies as well as an impurity test, drug release, and the uniformity of dosage unit are discussed from the viewpoint of international harmonization and technical advancement. PMID- 8717248 TI - [Improving on the Japanese pharmacopeia--on assays and determinations]. AB - Dr. Mitsuo Watanabe has offered many suggestions and points to be considered on the Japanese Pharmacopeia. His points of the matter include the nomenclature of reagents. Issues on electrodes and volumetric standard solutions to be used in nonaqueous potentiometric titrations are also involved, as well as the justification of a method for quantitative determination and analytical method validation. His concerns have prompted studies and careful surveys of these vital points to consider investigating that the nomenclature of reagents should be directed to prencipally follow the rules of IUPAC. Consideration has also been given to an issue of nonaqueous potentiometric titration, where problems many frequently be found with liquid junction between the electrode and the solution under titration. To work out the issue, investigations have started of an alternative technique characterized in using a platinum (or silver) electrode and a glass electrode as an indicator electrode and a reference electrode, respectively, without any liquid junction. For any titration system in question that can be regarded as nonaqueous titration from viewpoints of analytical chemistry. It has been suggested the use of any volumetric standard solutions of aqueous system should generally be avoided. Other points received consideration include sampling procedures for testing, interpretation of results from quantitation, and analytical method validation. PMID- 8717249 TI - [Globalization of herbal medicines]. AB - Herbal medicines, as the major remedy in traditional medical systems, have been used in medical practice and made a great contribution to maintaining human health. WHO is fully aware of the importance of herbal medicines of the health of many people throughout the world, and then WHO reported 3 kinds of guideline for herbal medicines. U.S.A. has the new bill for dietary supplement included herbal medicines, and been discussed the harmonizing North American Herbal Regulation. PMID- 8717250 TI - [Requirement for quality control of enzyme preparations]. AB - Enzyme preparations are medicine which prepared from natural and biotechnological enzymes or modified enzymes. The catalytic ability of the preparations display the efficacy of a medicine, and its chemical substance is protein. From these characteristics, the quality of enzyme preparations has to be evaluated at viewpoints what distinguish from synthetic drugs. Quality of enzyme preparations should be evaluated and assured from the following viewpoints: 1) definition of enzyme source, 2) identification of function for catalyst and substance as protein, 3) purity for functional enzyme protein, 4) biochemical evidence that justifies pharmacological action, 5) safety, and 6) contents. PMID- 8717251 TI - [Analysis of intracellular calcium concentration in tissues--special reference to synchronous analysis of the calcium and smooth muscle contraction]. PMID- 8717252 TI - [Diagnostic problems in clinical psychiatry]. PMID- 8717253 TI - [Aiming for the establishment of 10-year mental health planing]. PMID- 8717254 TI - [Learning from the Hanshin-Awajishima earthquake--report of the reality and future psychiatry in Japan]. PMID- 8717255 TI - [Caged compounds and measurement of calcium ion]. PMID- 8717256 TI - [Superior septal approach for mitral valve surgery]. AB - The superior septal approach (SSA) for mitral valve surgery is a useful approach, but it may bring about demage to sinus node function or an atrial electrical vulnerable condition because the sinus node artery is divided and a large portion of both atria is incised. From April 1992 to December 1994, we used this approach in 17 patients. There was no hospital death and none had perioperative complications associated with the approach. All nine patients in sinus rhythm preoperatively returned to sinus rhythm before discharge from the hospital and remained so at late follow-up. Five of 9 patients with sinus rhythm had electrophysiological examination preoperatively and all 9 patients had the examination postoperatively. Atrial vulnerability was not deteriorated after the operation in all cases. Under pharmacologic autonomic blockade, sinoatrial conduction time (SACT), sinus node recovery time (SNRT) after overdrive atrial pacing, and corrected SNRT were 73 +/- 35 msec, 1142 +/- 168 msec, and 365 +/- 122 msec, respectively. All these values were with in normal range and these were no significant differences between before and after the operation. All patients with sinus rhythm had 3 minutes of Bruce stage 1 exercise 9 months after the operation. The heart rate response to the exercise was as good as in healthy control people. In conclusion, as far as the midterm postoperative period, the effect of SSA on sinus node function and atrial vulnerability was acceptable. PMID- 8717257 TI - [Tricuspid valve closure for pulmonary atresia with intact ventricular septum]. AB - Tricuspid valve closure was performed on four children who had pulmonary atresia with intact ventricular septum. All of them had major right ventricle-coronary communications and severely hypoplastic right ventricle under 30% of normal right ventricle volume. Their coronary circulations were not right ventricle dependent. All of four patients survived tricuspid valve closure. Three children were performed Fontan procedure. One child, with severe change in her coronary arteries preoperatively, once improved her myocardial ischemia in early stage after tricuspid valve closure, however, the myocardial ischemia progrssed in one year later and died suddenly three years and five months after tricuspid valve closure. Three survivors remained good condition in their left ventricle function postoperatively. In this study, we investigated changes in LVEF, LVEDVI, LVEDP and mitral regurgitation as indicators of left ventricle functions through tricuspid valve closure. It revealed that tricuspid valve closure had advantages in preservation of left ventricle function and in preparation for Fontan procedure consequently. PMID- 8717258 TI - [Significance of hot shot in patients with unstable angina undergoing emergency coronary artery bypass graft surgery]. AB - We investigated the efficacy of terminal warm blood cardioplegia (hot shot) in patients with cardioplegic techniques. From January 1991 through April 1993, 68 patients recieved hypothermic cardioplegia induced by cold modified St. Thomas' Hospital solution containing 10mg/L of diltiazem hydrochloride followed by intermittent infusion of the cold crystalloid solution or cold blood cardioplegia without hot shot. From May 1993 through December 1994, 65 patients recieved hot shot before removal of aortic cross-clamp following the hypothermic cardioplegia. The earlier group consisted of 51 patients with stable angina undergoing elective CABG and 17 patients with unstable angina undergoing emergency of urgent CABG. The later group consisted of 44 patients with stable angina and 21 patients with unstable angina. The unstable angina undergoing coronary artery bypass graft surgery (CABG) using antegrade incidence of perioperative myocardial infarction was significantly higher in patients with unstable angina without hot shot (35%) compared to those with stable angina with and without hot shot (2% and 4%, respectively) and those with unstable angina with hot shot (5%). The level of maximum creatine kinase-MB (IU/L) was significantly greater in patients with unstable angina without hot shot (134 +/- 26) compared to those with stable angina with and without hot shot (57 +/- 7 and 65 +/- 4, respectively) and those with unstable angina with hot shot (57 +/- 8). The doses of dopamine and dobutamine (mg/kg) administered during 48 hours after CABG was not different between patients with stable angina with and without hot shot (13.9 +/- 0.9 vs 13.9 +/- 0.9), but tended to be lower in patients with unstable angina with hot shot (13.7 +/- 1.3) compared to those without hot shot (18.6 +/- 2.1). Left ventricular stroke work index (g.m/m2/b) immediately after CABG was comparable between patients with stable angina with and without hot shot (40 +/- 2.0 vs 36 +/- 1.5), but significantly greater in patients with unstable angina with hot shot (39 +/- 2.3) compared to those without hot shot (29 +/- 2.2). These results suggest that hot shot may provide a significant benefit in myocardial preservation during CABG especially in patients with unstable angina. PMID- 8717260 TI - [Expression of major histocompatibility complex of advanced non-small cell lung cancer (NSCLC)]. AB - Expression of major histocompatibility complex of lung cancer was examined to study the malignant potential of the tumor using immunochemical staining (anti HLA class I monoclonal antibody; w6/32). Nuclear DNA contents and the labeling index of proliferating cell nuclear antigen (PCNA) were measured as well. Forty three advanced (p-stage IIIa) lung cancers resected by lobectomy (33 cases), pneumonectomy (9 cases), and segmentectomy (1 case) entered the study. Mediastinal lymph node dissection was performed in every case during the operation. Of the 43 cases, expression of HLA class I was positive in 17 and negative in 26. The patients with positive HLA expression showed significantly better prognosis than those with negative HLA expression in terms of 5 year survival (p < 0.01). The patients with aneuploid pattern and positive HLA class I expression had the best prognosis on the survival curve (p < 0.01). When the PCNA positive rate was high, the prognosis was poor. And the patients with negative PCNA and positive HLA class I expression had significantly better prognosis than the others (p < 0.01). These findings suggest that the expression of HLA class I on the tumor cells is an important prognostic factor in the patients with NSCLC. PMID- 8717259 TI - [The experimental study of myocardial protection for warm myocardial ischemia in an isolated rat heart: the effect of a calcium antagonist]. AB - Although the protective effects of the calcium antagonists on ischemic and reperfused myocardium have been investigated, there have been only a few reports regarding their efficacy in relation to the degree of ischemic myocardium. This study was undertaken to investigate the efficacy of diltiazem, a calcium antagonist, in relation to the degree of ischemic myocardial injury in an isolated working rat heart. Three different models of ischemic injury were designed; Group A: 30 min global ischemia with a single dose infusion of St. Thomas' cardioplegic solution (STS), Group B: 60 min global ischemia with multidose infusion (every 30 min) of STS and Group C: 60 min global ischemia with multidose infusion (every 15 min) of STS. These groups received only STS, while Groups A-D, B-D and C-D (the treated groups) received the same solution with diltiazem (0.5 mumol/l). The recovery of post-ischemic cardiac function and the CPK leakage during reperfusion were evaluated, and the two groups were compared. For 30 min global ischemia, the addition of diltiazem to STS significantly improved the percentage recovery ratio of aortic flow (63.2 +/- 8.6% vs 79.9 +/- 5.9%, control vs. diltiazem, p < 0.01) and reduced CPK leakage during reperfusion (87.5 +/- 35.8 IU/20 min/g dry wt vs. 41.7 +/- 14.5 IU/20 min/g dry wt, control vs. diltiazem, p < 0.05). However, no differences in the post-ischemic functional recovery and CPK leakage were noted between the groups for 60 min global ischemia. In conclusion, for myocardial preservation, the addition of diltizaem to St. Thomas' cardioplegic solution was less effective for the 60 min global ischemia. Regarding severe myocardial ischemia, it was suggested that, inhibitation or suppression of calcium channel by diltizaem might insufficient to obtain additional protection of the St. Thomas' cardioplegic solution. Therefore, it would be necessary to control calcium entry through another pathway during ischemia and reperfusion. PMID- 8717261 TI - [Thoracoscopic surgery versus axillary thoracotomy for spontaneous pneumothorax]. AB - From October 1974 through March 1995, 453 patients with spontaneous pneumothorax admited to our institution and 416 patients underwent an operation. Among them, except bilateral pneumothorax, 204 patients under sixty years of age underwent an axillary incision (Group Ax) and 80 patients underwent a thoracoscopic surgery (Group Ts). In Group Ax, 178 male and 26 female, median age was 26.6 years old and in Group Ts, 68 male and 12 female, median age was 24.6 years old. The median operating time was significantly shorter for patients in Group Ts than for those in Group Ax (73.2 versus 111.3 minutes; p < 0.05). The median intraoperative blood loss was significantly less in Group Ts than in Group Ax (5.4 versus 65.6 g; p < 0.01). There was no significant difference in the duration of intercostal drain placement between the two groups (2.1 versus 2.7 days) but the median postoperative hospital stay was significantly shorter in Group Ts than in Group Ax (8.0 versus 12.5 days; p < 0.01). 58.9% of the patients needed postoperative analgesia in Group Ts, while 82.3% of the patients in Group Ax, and it was significantly less in Group Ts. The rate of recurrent pneumothorax was more in Group Ts (5.0% versus 2.9%), because of fail to notice the bulla under the thoracoscopy. Recently we observe the thoracic cavity using electric bronchoscope in order that we may not miss the bulla, and we take laser treatment for bullous emphysema with much effect. PMID- 8717262 TI - [Relationship between postoperative prognosis and preoperative immunological factors in aortitis syndrome]. AB - Valve detachment or pseudoaneurysm is a well known complication of surgical treatment for cardiovascular disease due to aortitis syndrome. Although the preoperative management of inflammation by steroid therapy is very important, occasionally, the operation in active phase might be recommended because of progressive heart failure or impending rupture of aortic aneurysm. The pathology of aortitis syndrome is related to immunological abnormality, but there has been little information concerning the immunological factors. In order to evaluate the relationship between immunological factors and surgical results in patients with aortitis syndrome, immunological examination was obtained before surgery in 12 patients. Cardiovascular lesions due to aortitis syndrome were aortic regurgitation in 6 patients, annuloaortic ectasia in 1 patient, aortic arch aneurysm in 1 patient, mitral regurgitation in 2 patients, and coronary artery disease in 2 patients. Hospital mortality including initial operative death was not found. However, 6 late death (50%) occurred at late time with the mean of 23.8 months because of severe complications such as valve detachement or pseudoaneurysm after operation. In a comparison of the preoperative immunological values between survival group and late death group, immunoglobulins such as IgG, IgA, C3 and C4 were significantly higher in late death group. According to the recurrence of inflammation, the patients were divided into three groups. Group A included 3 patients who remains in active phase after initial operation in active phase. Group B included 3 patients who changed to active phase after initial operation in inactive phase. Group C included 6 patients who remains in inactive phase after initial operation. In a comparison among 3 groups, IgG and C4 showed no significant difference. However, IgA and C3 of group C were significantly lowest among three groups. This outcome is considered to be related to existence of latent inflammation. At operation, insertion of rigid prosthesis to fragile position due to latent inflammation may stimulate immunological reactions. We conclude that the preoperative values of the immunological factors, especially IgA and C3, are reliable predictors in postoperative prognosis. PMID- 8717263 TI - [Basic amino acid, L-arginine aggravates ischemia-reperfusion injury]. AB - Basic amino acid, L-arginine has been known to have several biological actions including nitric oxide production. The real effects caused by L-arginine administration to heart has not been fairly understood in the setting of ischemia reperfusion process. The effects of L-arginine on recovery of myocardial contractile function and oxidative metabolism were studied in a model of reversible global normothermic ischemic injury using an isolated, buffer-perfused rabbit heart preparation. 1 mM L-arginine or vehicle was infused into hearts for 2 minutes at the onset of 35 minutes of ischemia in L-arg. group or in control group, respectively. Oxygen consumption (MVO2), lactate release into coronary vessels, and cardiac function including developed pressure, +dp/dt, -dp/dt and diastolic pressure-volume relationship (PVR) were measured and high energy phosphates were also evaluated by 31P-NMR spectroscopy. Endothelial function was also evaluated by acetylcholine (Ach) infusion with monitoring of constant perfusion pressure. L-arginine caused a significant increase in the PVR and profund decline in systolic function when compared to control group. MVO2 was significantly impaired to cause a decrease of high energy phosphates (phosphocreatine and ATP). Lactate release into coronary vessels on reperfusion was significantly higher in L-arg group than that in control group, suggesting a promotion of anaerobic glycolysis. Deterioration of endothelial function and smooth muscle function of artery were evidenced by Ach infusion test. Although the mechanisms of injury are speculatory, possible mechanisms of this injury are stimulation of nitric oxide production by L-arginine and cation change, especially calcium accumulation. We concluded that L-arginine was able to cause aggravation of ischemia-reperfusion injury with reduced contractile function and mitochondrial function and increased anaerobic glycolysis after ischemia reperfusion in an isolated model of reversible ischemic injury. PMID- 8717264 TI - [Evaluation of the prognosis of patients with stage I non-small cell lung cancer with respect to predicted postoperative lung function]. AB - One hundred and forty patients underwent absolute curative resction for stage I non-small lung cancer from 1982 to 1993 at our department. For these, prognosis and changes in quality of life (QOL) were evaluated retrospectively with respect to the predicted postoperative lung function. The average age of the patients was 62 years (range 31 to 84 years), and 103 males and 37 females were included. Seventy-five of the patients had adenocarcinoma, 61 squamous cell carcinoma, and 4 large cell carcinoma. These 140 patients were classified into two groups, H and N, according to the predicted postoperative %FEV1.0 and %VC. Group H patients (n = 39) had a predicted %FEV1.0 and/or %VC of 55% or less for postoperative respiratory disease. Group N patients (n = 101) had a predicted %FEV1.0 and %VC of 56% or more for expected normal respiratory conditions postoperatively. Group N patients showed a 98% one-year survival rate, and 72% five-year survival rate and good QOL postoperatively. On the other hand, group H patients showed 86% and 45% one- and five-year survival rates respectively, the same as those predicted for patients with stage II non-small cell lung cancer. Furthermore, group H patients more than 70 years old showed 80% and 17% one- and five-year survival rates, the same as those predicted for patients with stage IIIA non-small cell lung cancer, and poor prognosis in comparison with that of the group N patients more than 70 years old. Then, QOL was investigated one year postoperatively. The group H patients showed deterioration of performance status in comparison with the group N patients. Since there was a high incidence of postoperative respiratory disease in the patients with a predicted %FEV1.0 and/or %VC of 55% or less, physicians should avoid extended lung resection, which might cause deterioration of the cardiopulmonary reserve volume, in patients in whom respiratory disease is predicted to occur, particularly for patients more than 70 years old. In conclusion, physicians should consider limiting surgical intervention to preserve lung volume and QOL of patients more than 70 years old with restricted cardiopulmonary function. PMID- 8717265 TI - [Clinical study of nineteen thymic carcinomas]. AB - Nineteen cases of thymic carcinoma treated in our hospital (mean age 60.0 years, seven males and twelve females) were studied clinically. Thirteen cases (68.4%) had subjective symptom; for example, chest pain, face edema or cough. Two cases (10.5%) had the associated diseases; one had gammaglobulinemia, the other one had Cushing syndrome and hypogammaglobulinemia. The histological subtypes were eleven squamous cell carcinomas (SCC) (57.9%), three undifferentiated carcinomas, two small cell carcinomas, one papillary adenocarcinoma and one lymphoepithelioma like carcinoma. The 5 year survival rate of all cases was 42.7%. The 10 years survival rate was 21.4%. The median survival time was 56.1 months. There were one Stage I case, nine stage III cases, four stage IVa cases and five stage IVb cases. More than stage III cases were eighteen (94.7%). Five cases of stage IVb were T2N1M0, T3N1M0, T3N0M1 and T4N0M1. The 5 year survival rate of SCC was 65.6%, and that of the other subtypes was 14.3%. The cases resected completely were only eight cases (42.1%), and the 5 year survival rate of these was 70.0%. On the other hand, the rate of five cases resected incompletely was 53.3%. Metastasis occurred in 12 cases (63.2%). Metastasis occurred frequently in pleura, lymphnode, lung, bone and liver. The radiotherapy for SCC was effective and the 5 year survival rate was 83.3%. Furthermore, there were some long survivors in the cases undergone incomplete resections by postoperative radiotherapy jointly. On the other hand, the chemotherapy was not effective in our series. However, it was reported recently that the regimen including cisplatin was effective. So it was impressed that the combined therapy including surgery radiotherapy and induction chemotherapy would be important to obtain better results. PMID- 8717266 TI - [Comparison between retrograde cerebral perfusion and selective cerebral perfusion by intraoperative electroencepharogram]. AB - We used retrograde cerebral perfusion (RCP) to protect the brain in patients undergoing aortic arch replacement from May 1991 to July 1992. Subsequently, we have been using selective cerebral perfusion (SCP). In this study, a group of six patients undergoing RPC were compared with 5 patients undergoing SCP with regard to the intraoperative electroencepharogram (EEG) and post operative clinical course. There were no deaths in either group, but transient palsy of the right arm developed in one patient in the RCP group. The amplitude of intraoperative EEG was examined at four points: before cardiopulmonary bypass, during RCP or SCP, at 30 degrees C and at 35 degrees C during rewarming. The EEG amplitude at these four points was expressed as a ratio to the value before cardiopulmonary bypass. The ratios at these four points were 1, 0, 0.08, and 0.19 in the RCP group and 1, 0, 0.70, and 0.90 in the SCP group, respectively. In the SCP group, EEG recovered rapidly, whereas in the RCP group EEG did not recover until the completion of cardiopulmonary bypass in all but one patient. These results indicated that RCP does not provide reliable protection of the brain. PMID- 8717267 TI - [Case of benign metastaxsizing leiomyoma of the lungs]. AB - A 51-year-old woman with bilateral multiple pulmonary metastases of uterine leiomyoma was reported. The patient had undergone a hysterectomy for uterine myoma in 1976. In 1981, routine chest roentgenograms showed three nodular lesions in the right lung with well-defined margins. The patient received no treatment because she was asymptomatic. In January 1993, she consulted a gynecologist because of acute lower abdominal pain and an abdominal mass, and bilateral pulmonary tumors were found. The retroperitoneal solid tumor was extirpated in the first operation, and histological findings showed that it was a benign leiomyoma. The pulmonary tumors, extirpated in the second operation, were benign leiomyoma, also. The clinical course and histological findings suggested that this was a case of benign metastasizing leiomyoma. This disease is rare, 21 case have been reported in Japan. PMID- 8717268 TI - [Reoperation of a pseudoaneurysm eroding the sternum under hypothermic arrest of the brain through left thoracotomy]. AB - Reoperation of a pseudo aneurysm eroding the sternum was carried out in a 76-year old female, using partial cardiopulmonary bypass and hypothermic circulatory arrest of the brain through left thoracotomy. The pulsating aneurysm protruding over the sternum was about 3.5 x 3.5 cm in size. Therefore, median sternotomy was thought to be risky. Cardiopulmonary bypass was established with left atrium and pulmonary artery for venous cannulation and femoral artery for arterial one. The patient was cooled and then the bypass discontinued. Previous patcth of the aneurysm was partially splitting and new patch was sutured. She awaked without cerebral injury of the operative day. This technique can be used as one of an alternative surgical method in the treatment of the distal aortic arch aneurysms. PMID- 8717269 TI - [Case report of one pediatric patient surviving 6 years after heart transplantation in the U.K]. AB - In October 1988, a six year old Japanese girl with restrictive cardiomyopathy underwent heart transplantation (HTx) at Harefield Hospital in the U.K. Six years 9 months after her HTx, she is doing well and is the longest survivor among the Japanese HTx recipients. Cardiac catheterization 5 years after her HTx showed good cardiac function with LVEF of 54.7%. Coronary angiogram at that time showed no abnormal findings. Cardiac biopsy revealed no sign of rejection or myocyte degeneration. LVDd (Left Ventricular Diastolic Dimension) grew from 35.5 mm at the time of transplant to 42.6 mm 5 years later by echocardiogram, suggesting the growth of the transplanted heart. Her good progress, including catch-up growth and good QOL, reflects the excellent benefit of HTx today. The authors deeply appreciate the courtesy of Harefield Hospital in accepting to perform her HTx. PMID- 8717270 TI - [Intrathoracic transposition of the muscle flap of the trapezius and rhomboideus in treating empyema]. AB - Intrathoracic transposition of the muscle flaps of the trapezius and rhomboideus was performed simultaneously with an open-drainage thoracotomy in a 66-year-old male with empyema. These muscle flaps obliterated the upper empyema cavity and subsequently made it easier for obliteration the remaining lower empyema cavity. The muscle flaps of the trapezius and rhomboideus are useful in obliteration the upper empyema cavity. PMID- 8717271 TI - [Giant isolated extracadiac unruptured aneurysm of the right coronary sinus of valsalva--a case report]. AB - A giant isolated extracardiac unruptured aneurysm of the right coronary sinus of Valsalva was detected incidentally in a 24-year-old man with clinical findings of Ehlers-Danlos syndrome. The right coronary artery was occluded at its ostial resion. Surgical correction was performed by obliterating the orifice of the aneurysm with a Dacron patch. Microscopic examination of the diseased aortic wall revealed absence of medial elastic fibers. Collective review of 19 reported similar cases revealed that reports concerning unruptured, isolated giant sinus of Valsalva aneurysm were rare, especially in young patients, and that the aneurysm of this patient was the largest. PMID- 8717272 TI - [A case report of a bronchogenic cyst of the diaphragm causing rupturing into the left pleural cavity]. AB - A rare case of a bronchogenic cyst of the diaphragm was reported. The patient was a 56-year-old woman complaining of epigastric pain. A CT scan and a MRI examination revealed a cystic tumor at the left vertebro-phrenic angle. The cyst ruptured into the left pleural cavity causing chest pain and consequently reducing its size. As the cyst had enlarged again after 6 months, she received surgical treatment and the tumor was totally removed. The cyst was monolocular, originating from the diaphragm and proved to be bronchogenic cyst. It had bronchial epithelium, smooth muscle, bronchial gland and cartilage histologically. Japanese literature on the bronchogenic cysts of the diaphragm were discussed. Seven cases, including this case, of bronchogenic cysts of the diaphragm have been reported in Japan. It is rare for bronchogenic cysts to rupture into the pleural cavity. PMID- 8717273 TI - [Axillo-iliac bypass grafting for reoperation of graft stenosis after aortic arch reconstruction of type B interruption of the aortic arch: two cases report]. AB - Right axillo-iliac bypass grafting was performed in two females (10 and 14 years old) who had stenosis or obstruction of grafts after reconstruction of the aortic arch in type B interruption. The initial bypass operations were carried out at the age of 5 months and 5 years, with the use of a 5-mm EPTFE graft and a 10-mm Dacron graft. Nine years later, the EPTFE graft was completely obstructed, and the Dacron bypass has stenosis, kink and calcification. To minimize surgical invasion, axillo-iliac extra-anatomical bypass was employed in both patients. 8 mm or 10-mm Hemashield grafts were implanted between the right axillar artery and the right common iliac artery through the intrapleural-preperitoneal route, and favourable results were obtained in both patients. The axillo-iliac bypass is considered to be a beneficial conservative method of reoperation for stenosis after graft reconstruction of the aortic arch in the younger age patients. PMID- 8717274 TI - [Anomalous origin of the left coronary artery from the pulmonary artery coronary revascularization under normothermic coronary perfusion in an infant]. AB - A 5-months-old baby was hospitalized with congestive heart failure. Anomalous origin of the left coronary artery from the pulmonary artery was diagnosed on echo cardiography and coronary angiogram. Preoperative examination revealed severe left ventricular dysfunction (left ventricular end-diastolic pressure was 32 mmHg, left ventricular ejection fraction was 16%). Coronary revascularization was performed by anastomosis of the neo-left coronary artery to the aorta using an autologous pulmonary arterial roll under normothermic coronary perfusion. Although postoperative CPK-MB was within normal limits, the patient died on the 12th postoperative day due to ventricular arrythmia. Autopsy revealed severe endomyocardial fibroelastosis. PMID- 8717275 TI - [Radiation-induced coronary ostial stenosis, a case of redo coronary bypass for the restenosis following patch angioplasty]. AB - The patient was a 45-year-old woman who had had a tumor resection for thymic carcinoid and subsequent mediastinal irradiation (50 Gy) 3 years before the onset of angina pectoris during exercise. Coronary angiography (CAG) revealed an isolated ostial stenosis of the left main coronary trunk (LMT). Angiography also showed an occluded right internal thoracic artery (ITA) at its origin. The patient underwent patch angioplasty of the LMT orifice using a piece of the saphenous vein graft (SVG). One month after the operation, CAG revealed a success of operation with an enlarged LMT orifice and she was discharged. However, 3 months after the operation, angina pectoris recurred and a repeated CAG showed a 90% stenosis of the LMT at the place 1 cm distal to the orifice. Emergency CABG (the left ITA to the LAD and the SVG to the LCX) was accomplished with disappearance of angina. Post-CABG angiography revealed patent left ITA and SVG in association with complete obstruction of the LMT. One year after the second operation, she was free from symptoms. This case as well as other reports concerning radiation-induced coronary stenosis suggest that patch angioplasty for this specific lesions may have a high incidence of stenosis recurrence. Coronary bypass grafting that can be performed at the place away from the active proliferative lesion may be a better selection. PMID- 8717276 TI - [Comparative biochemical studies of cholanoids]. AB - Bile acids and bile alcohols are termed cholanoids. Biles of evolutionarily primtive vertebrates such as fishes, amphibians, and reptiles contain bile alcohols and higher bile acids in place of C24 bile acids in mammals. These higher cholanoids have the C24 bile acid type of nuclear structure and all or part of the side chain of cholesterol. The chemical structure, the natural distribution, and the biosynthetic pathways of bile alcohols and higher bile acids were studied. The results indicate that these compounds are evolutional precursors of the C24 bile acids found in mammalian species and the mechanism of the conversion of cholesterol to the C24 bile acids in mammals is a recapitulation of the evolution of cholanoid molecules. PMID- 8717277 TI - [Genes for gastric proton pump and their transcriptional regulation]. AB - The highly differentiated gastric parietal cell has a characteristic morphology and plays a specialized role in the hydrochloric acid secretion into the stomach lumen. The major enzyme in this system is an ATP-driven proton pump, the H+/K(+) ATPase, which is responsible for proton translocation across the apical plasma membrane. The primary structures of the catalytic alpha and glycosylated beta subunits, and their transmembrane topology are similar to those of the corresponding subunits of Na+/K(+)-ATPase, suggesting that the reaction mechanism of both ATPases would be essentially the same if not identical. Most of the positions of introns in the H+/K(+)-ATPase subunit gene. These findings suggest that the alpha and beta subunit genes, respectively, of the two ATPases were derived from the common ancestors. We found that a DNA sequence motif, (G/C)PuPu(G/C)NGAT(A/T)PuPy, was located in the upstream regions of both alpha and beta subunit genes from human and rat. This motif may be a binding site for a positive transcriptional regulator that functions specifically in the parietal cells. cDNA cloning and in situ hybridization demonstrated that novel zinc finger proteins (GATA-GT1 and GATA-GT2) are present in the gastric parietal cells. These proteins bind to the (G/C)PuPu(G/C)NGAT(A/T)PuPy motif. Furthermore, they activate the transcription of the reporter gene with the 5'-upstream region of the alpha or beta subunit gene. These results suggest that gastric GATA DNA binding proteins play important roles in transcriptional activation of H+/K(+) ATPase genes in the parietal cells. PMID- 8717278 TI - [New intracellular calcium antagonists. I. Synthesis and pharmacological evaluation of 2,3,4,5-tetrahydro-1,5-benzothiazepine analogs]. AB - A series of 2,3,4,5-tetrahydro-1,5-benzothiazepine and related compounds were prepared, and the intracellular Ca2+ inhibitory effects were examined using methoxamine- or caffeine-induced contraction of isolated rabbit arteries. Structure-activity relationship studies of these compounds are discussed and the results suggest that novel 5-[3-[2-(3,4-dimethoxyphenyl)ethyl]aminopropionyl] 2,3,4,5- tetrahydro-1,5-benzothiazepine fumarate (20d) showed the most potent inhibitory action on the intracellular Ca2+ release. PMID- 8717279 TI - [Pharmacognostical studies on adiantum plants. V. Classification based on spore morphology and distributional patterns of silicon and calcium in the ultimate pinnules]. AB - Crude drugs derived from Adiantum species are used as febrifuge, antidote, diuretic, tonic, etc. Some commercial samples of these drugs are composed of finely cut ultimate pinnules only, which have false indusia and spores. In this paper, in order to establish a classification method based on the characteristics of the ultimate pinnules and their attachments, the morphological study using stereoscope and scanning electron microscope, and X-ray microanalysis using an electron probe microanalyzer were carried out on the false indusia and spores, and the ultimate pinnules, respectively, of 19 Adiantum species. The results showed that examined all species could be distinguished from each other by the following characteristics: in the false indusium, the shape and the presence or absence of hairs; in the spore, the shape, the ornamentation, the ratio of the laesura, and the equatorial diameter; in the X-ray images of the ultimate pinnule, the distributional patterns of silicon and calcium. The distributional patterns of silicon were due to the presence of spicular cells, hairs and papillae, and calcium was present as crystals of calcium oxalate. The average content of silicon in the ultimate pinnules of Adiantum species was 1.99%. PMID- 8717280 TI - [Quality evaluation on Paeoniae radix. I. Quantitative analysis of monoterpene glycosides constituents of Paeoniae radix by means of high performance liquid chromatography. Comparative characterization of the external figures, processing method and the cultivated areas]. AB - The constituents of monoterpene in Paeoniae Radix were analyzed by means of high performance liquid chromatography (HPLC) using hitherto elucidated eight monoterpene glycosides, paeoniflorin 1, oxypaeoniflorin 2, benzoylpaeoniflorin, 3, benzoyloxypaeoniflorin 4, galloylpaeniflorin 5, galloyloxypaeoniflorin 6, albiflorin 7 and lactiflorin 8 and a monoterpene paeoniflorigenone 9. In sixty seven kinds of Paeoniae Radix collected in Japan, China, and South and North Korea those monoterpene constituents 1-9 were found to exist in the amounts of ranging 0.12-9.61% 1, 0.06-10.8% 2, 0.02-0.79% 3, 0.24-0.47% 4, 0.25-2.53% 5, 0.05-2.86% 6, 0.09-2.76% 7, 0.35-0.64% 8 and 0.01-0.49% 9 respectively. In addition, these results obtained by the quantitative analysis were discussed from the viewpoint of the external figures and processing methods. PMID- 8717281 TI - [The constituents of Taxillus kaempferi and the host, Pinus thunbergii. I. Catechins and flavones of Taxillus kaempferi]. AB - Four flavonoid compounds, avicularin (1), hyperin (2), quercitrin (3), taxillusin (4), were isolated from leaves of Taxillus kaempferi, and three catechin compounds, (+)-catechin (6), procyanidin B-1 (7), procyanidin B-3 (8), showing a reddish purple color on the reaction with Ehrlich reagent, were also isolated from the stems. Each of these seven isolated compounds were identified by comparing the spectral data with those of the references. Compound 8 was examined to substantiate the stereochemical configurations of conformer, a and b in the detailed 1H-nuclear magnetic resonance (1H-NMR) spectrum using the nuclear Overhauser effect (NOE) difference spectroscopy and spin decoupling experiment in the acetone-d6 solution. PMID- 8717282 TI - [Studies on Rehmanniae radix. III. The relation between changes of constituents and improvable effects on hemorheology with the processing of roots of Rehmannia glutinosa]. AB - The relation between constituents and effects on hemorheology with processing of root of Rehmannia glutinosa was investigated. With the processing to dried or steamed root from crude root, the content of stachyose, which was the highest component in crude root, was decreased and the content of manninotriose was increased. Iridoid glycoside, catalpol, was gradually decreased with the processing. From these results, the content of carbohydrates and catalpol made it possible to estimate the quality of Rehmanniae Radix by processings. On the other hand, the change of pharmacological activities with the processing was investigated. The 50% ethanolic extract (BJ-ext) from steamed root of R. glutinosa increased erythrocyte deformability and erythrocyte ATP contents, inhibited polybrene-induced erythrocyte aggregation, and promoted an activity of fibrinolytic system. The extracts from crude or dried root had weak or no effect. The crude or dried root and steamed root of R. glutinosa showed the different pharmacological activities, and the quality of Rehamanniae Radix by processings may be estimated by investigation of correlation between the changes of constituents and improvable effects of hemorheology. PMID- 8717283 TI - [Influences of a shiitake (Lentinus edodes)-fructo-oligosaccharide mixture (SK 204) on experimental pulmonary thrombosis in rats]. AB - Effects of the mixture (SK-204) consisting of dried shiitake mushroom (Lentinus edodes) treated with wet-heating and fructo-oligosaccharides (7:3) on the experimental models of pulmonary thrombosis induced by lactic acidosis in rats were evaluated. Chronic oral administration (10 weeks) of SK-204 significantly prevented the thrombus formation on this thrombosis model. However, decreases in the numbers of platelet and fibrinogen level by lactate were not changed by SK 204. These results suggest that SK-204 have an anti-thrombotic action, which is due to neither the inhibition of platelet aggregation nor coagulation, but probably due to the promotion of fibrinolysis and thrombolysis. PMID- 8717284 TI - [DNA strand breaks in epithelial cells from mice with bleomycin induced pulmonary fibrosis]. AB - Bleomycin-induced cytotoxicity is believed to be caused by single- and double strand DNA breaks. To examine the effect of bleomycin on DNA strand breaks and the role of these breaks in bleomycin induced pulmonary fibrosis in mice, we analyzed DNA strand breaks in situ by TdT-mediated dUTP-biotin nick end labeling (TUNEL), previously described by Gavrieli et al. The nuclei of bronchiolar epithelial cells were strongly stained 1 hr to 12 hr after bleomycin administration, and after that period DNA damage was repaired. Nuclei of alveolar epithelial cells showed positive signals correlated with progression of fibrosis. Although corticosteroids did not block the early DNA damage in bronchiolar epithelial cells, they did inhibit later damage to alveolar epithelial cells and fibrosis. We speculate that the DNA damage in alveolar epithelial cells and the progression of fibrosis in later stages are associated with inflammatory cytokines. These findings show the location and the time course of the DNA damage in bleomycin-induced pneumonitis in mice, and they indicate that the prolongation of DNA damage in alveolar epithelial cells is closely related to fibrinogenesis. PMID- 8717285 TI - [Immunohistochemical study of fibrosis and S-100 protein-positive cells in idiopathic pulmonary fibrosis and in interstitial pneumonia associated with rheumatoid arthritis]. AB - Immunohistochemical methods were used to distinguish idiopathic pulmonary fibrosis (IPF) from interstitial pneumonia associated with rheumatoid arthritis (RA lung). The subjects were six patients with IPF and seven with RA lung, in whom the pathological findings were consistent with usual interstitial pneumonia. Antibodies to vimentin (Vim), alpha-smooth muscle actin (alpha-SMA), and S-100 protein were used for immunohistochemical studies done by the streptavidin-biotin peroxidase complex method. In fibrosis associated with RA lung, proliferation of both Vim and alpha-SMA-positive myofibroblasts was widely observed, despite pathological findings of honeycombing, usual interstitial pneumonia, and BOOP. Fibrosis in cases of IPF was found to be characterized mainly by Vim-positive fibroblasts, and on occasion was associated with hyperplasia of smooth muscle. Lung tissues from patients with acute exacerbations of RA lung, especially when associated with a BOOP pattern, had many cells positive for S-100 protein. However, such cells were generally hard to find in cases of IPE. Similar results were obtained with regard to the honeycomb pattern in both IPF and RA lung. These findings suggest that IPF and RA lung can be fairly clearly differentiated based on the proliferation of myofibroblasts and on the presence or absence of cells positive for S-100 protein. PMID- 8717286 TI - [Effects of substance P and neurokinin A on pulmonary circulation in the isolated perfused guinea pig lung]. AB - We studied the effect of sensory nerve peptide substance P (SP) and neurokinin A (NKA) in isolated perfused guinea pig lungs. SP and NKA increased pulmonary arterial pressure, capillary pressure, pulmonary venous resistance, and lung weight, but they did not change pulmonary arterial resistance or pulmonary vascular permeability. The effects of SP on pulmonary vascular dynamics were greater than those of NKA. Ozagrel hydrochloride, a thromboxane synthase inhibitor, partially attenuated the effect of SP. This indicates that thromboxane contributes to SP-induced pulmonary vasoreactivity. However, ozagrel hydrochloride did not change the effects of NKA. FK224, an NK-1/NK-2 receptor antagonist, abolished both SP- and NKA-induced pulmonary vasoconstriction. This indicates that SP and NKA acted on the pulmonary vasculature through the NK-1 or NK-2 receptor, or both. Papaverine, a smooth muscle relaxant, abolished the effects of SP. The SP-induced increase in lung weight was caused by a rise in pulmonary hydrostatic pressure, especially that caused by pulmonary venoconstriction. PMID- 8717287 TI - [Eosinophil cationic protein in serum from adults with asthma and with chronic obstructive pulmonary disease]. AB - Levels of eosinophil cationic protein (ECP) in serum are abnormally high in bronchial asthma, and may reflect the degree of eosinophilic inflammation in the lung. The purposes of this study were to evaluate whether serum ECP levels are more closely related to the severity of asthma than are peripheral blood eosinophil counts, and whether these levels can be used to distinguich bronchial asthma from pulmonary emphysema and chronic bronchitis. Serum ECP levels were measured and blood eosinophil counts were obtained in 46 symptomatic adults with asthma (31 atopic and 15 non-atopic), 10 patients with pulmonary emphysema, 11 patients with chronic bronchitis, and 30 non-smoking healthy subjects. Both serum ECP levels and blood eosinophil counts were higher in patients with asthma than in those with pulmonary emphysema, chronic bronchitis and in healthy subjects. Neither serum ECP levels nor blood eosinophil counts differed between atopic and non-atopic patients with asthma. After therapy, serum ECP levels were found to have decreased to have decreased significantly (p < 0.025), but blood eosinophil counts were not. We conclude that both serum ECP levels and blood eosinophil counts are useful for distinguishing bronchial asthma from chronic obstructive pulmonary diseases. These data also indicate that in adults with asthma serum ECP levels may be more closely related to clinical severity than are blood eosinophil counts. PMID- 8717288 TI - [Radiologic-pathologic correlation between tumor edge and surrounding inherent structures in peripheral lung cancer]. AB - PURPOSE: To evaluate the radiologic-pathologic relationship between the tumor edge of peripheral lung cancers and surrounding normal structures. MATERIALS AND METHODS: Tissue samples from 16 lung cancers with diameters less than 2 cm were examined: 7 adenocarcinomas, 5 squamous cell carcinomas, 2 small cell carcinomas, 1 large cell carcinoma, and 1 that had metastasized to the lung. Inflation-fixed lung specimens containing tumor tissue were sliced to a thickness of 2 mm: The tumor edge of each section was traced under a stereomicroscope and its relationship to marginal structure, including bronchi, pulmonary arteries, and pulmonary veins containing interlobular septa, was analyzed. The tumor edge and marginal structures were classified into four types: Type 1, clear margin formed by marginal structures; Type 2, tumor involving the marginal structures; Types 3 and 4, marginal structures penetrating the tumor with and without a notch, respectively. RESULTS: Type 1 was found in 23 of 47 pulmonary arteries (49%), 23 of 49 bronchi (47%), and 12 of 31 pulmonary veins or interlobular septa (39%). Type 2 was found in 11% (14/127), Type 3 in 15% (19/127) and Type 4 in 28% (36/127) of marginal structures. Type 1 was common in all histological types of carcinomas studied. CONCLUSION: In small peripheral lung cancers, the tumor edge sometimes has a clear margin formed by surrounding structures such as interlobular septa. This can make it difficult to differentiate benign from malignant lesions. PMID- 8717289 TI - [Effects of thromboxane A2 on chronic hypoxic pulmonary hypertension in the rat]. AB - Chronic hypoxia (10% O2 for 2-3 weeks) causes pulmonary hypertension and vascular remodeling in the rat. To study the role of thromboxase A2 in chronic hypoxic pulmonary hypertension, the hemodynamic effects of intravenous administration of a thromboxane analogue (STA2) were measured in chronic hypoxic (H) and normoxic (N) Sprague-Dawley rats. During anesthesia baseline pulmonary arterial pressure (PAP) was higher in H rats (34.6 +/- 1.0 mmHg) than in N rats (18.4 +/- 1.2 mmHg). Intravenous STA2 (0.3 microgram) acutely increased pulmonary artery pressure by 74% +/- 11% (25 +/- 4 mmHg) in H rats and by 47% +/- 2% (9 +/- 1 mmHg) in N rats, which indicates that both the absolute and relative acute pulmonary vasoconstriction caused by STA2 were greater in H rats. The changes in systemic arterial pressure caused by STA2 were smaller than the changes in pulmonary arterial pressure both in H rats (11% +/- 3%) and in N rats (17% +/- 3%). Lungs were isolated and perfused with saline, and the vasoconstrictive response to 0.05 microgram of STA2 in lungs (14.5 +/- 2.4 mmHg) from H rats was greater than the response to 0.1 microgram of STA2 (5.6 +/- 1.3 mmHg) in lungs from N rats. To examine whether blockade of calcium channels could suppress the vasoconstrictor response to STA2, the effects of the calcium channel blocker nicardipine hydrochloride on vasoconstriction caused by STA2 were measured in H and N rats. In vivo, the blockade of calcium channels suppressed the increase in pulmonary artery pressure caused by STA2. This suppression was greater in H rats (56% +/- 11%) than in N rats (25% +/- 4%). Similar results were obtained with isolated perfused lungs. Blockade of calcium channels suppressed the vasoconstriction caused by STA2 and this suppression was greater in H rats than in N rats. The finding that thromboxane A2 induced greater vasoconstriction in H rats than in N rats indicates that thromboxane A2 may play an important role in pulmonary hypertension, and suggests that blockade thromboxane A2 may benefit some patients with primary and secondary pulmonary hypertension. Furthermore, the finding that suppression of thromboxane-induced vasoconstriction by blockade of calcium channels was greater in H rats than in N rats indicates that such treatment may also benefit some patients. PMID- 8717290 TI - [Compliance with long-term home oxygen therapy]. AB - Long-term home oxygen therapy has been shown to benefit patients with hypoxemic chronic obstructive pulmonary disease. However, to obtain the expected maximal benefit it is important for the oxygen to be used correctly and for a sufficient length of time. We examined compliance with home oxygen therapy in patients with chronic obstructive pulmonary disease, pulmonary fibrosis, late sequelae of pulmonary tuberculosis, and pulmonary hypertension who used oxygen concentrations. Compliance was defined as the ratio of the amount of oxygen used to the amount prescribed. The average daily length of time the concentrator actually ran was measured from the concentrator meters. These were read every 6 months by an engineer from the company that installed the concentrator. Factors thought to affect compliance were studied. These factors included age, the degree of dyspnea, arterial blood gases, and pulmonary function. Weak positive correlations were found between compliance and age and between compliance and PaCO2. A weak negative correlation was observed between compliance and PaO2. Compliance in patients with chronic obstructive pulmonary disease was higher than in patients with pulmonary fibrosis or pulmonary hypertension. Among those given prescriptions for 24-hr oxygen therapy, compliant patients had more severe dyspnea on excertion than did noncompliant patients. These data suggest that the compliant patients had more severe gas exchange problems. PMID- 8717291 TI - [Effects of subacute hypoxia on alveolar epithelial ion transport in rats]. AB - To study the effects of subacute hypoxia on alveolar epithelial ion transport, alveolar fluid clearance was measured in isolated fluid-filled rat lungs. After instillation of a solution containing about 5% bovine albumin, an increase in alveolar fluid clearance was measured over 2 hours. Alveolar fluid clearance was lower when rats were kept in hypoxic conditions (FiO2 = 0.1) for 48 hours. Neither 10(-5) M amiloride (a Na(+)-channel blocker) nor 10(-3) M ouabain (an inhibitor of Na(+)-K(+)-ATPase) inhibited fluid clearance in the hypoxia group, but they did in the normoxia group. These results indicate that subacute hypoxia may down-regulate both the Na(+)-channl and Na(+)-K(+)-ATPase, and thus decrease the absorption of excess alveolar fluid. PMID- 8717292 TI - [Middle lobe syndrome--incidence and relationship to atypical mycobacterial pulmonary disease]. AB - We evaluated the incidence of middle lobe syndrome in the Haibara area, and its relationship to atypical mycobacterial infection. Of the 30,588 persons who underwent annual mini-chest roentgenography in 1992 or 1993 or both, 51 (0.17%) had middle lobe syndrome, diagnosed from posteroanterior and lateral chest X-ray films. The incidence was significantly higher in persons over 50 years old than in persons under 50 years old (0.26% vs 0.02%: p < 0.001), and was higher in femals than in males (0.20% vs 0.11%: p = 0.527). Of 16 patients examined by bronchoscopy and computed tomography, 7 showed evidence of cylindrical bronchiectasis, and four had mycobacterium avium complex pulmonary disease presenting as middle lobe syndrome. All four were women who were 51 years of age or older and none had predisposing pulmonary disorders. Computed tomography showed multiple nodular shadows with or without bronchiectasis located in the middle lobe or the lingula. Cavitary lesions were not seen. These results indicate that middle lobe syndrome is not rare, and that infection with mycobacterium avium complex should be considered when multiple nodular shadows are seen in the middle lobe or the lingula. PMID- 8717293 TI - [Pneumothorax from a nasogastric feeding tube]. AB - An 83-year-old woman was admitted to our hospital because of fever and disturbance of consciousness. Pneumonia and multiple organ failure were diagnosed from symptoms, a chest X-ray film, and laboratory findings. With the administration of antibiotics and an antiprotease inhibitor, the pneumonia and multiple organ failure began to resolve, but consciousness remained disturbed. A nasogastric feeding tube was inserted to provide parenteral nutrition. Neither abnormal resistance nor reflex coughing were noted during insertion. Bubbling rales were not heard on auscultation, and a chest X-ray film revealed that the tube had traversed the right bronchial tree, perforated the lung, coiled up in the right pleural vavity, and caused a right pneumothorax. The tube was immediately removed. The pneumothorax was treated with an intercostal drainage tube for 1 day and resolved without further problems. This case shows one of the dangers involved in inserting nasogastric feeding tubes. The medical community should be aware that pulmonary complications may develop after such tubes are inserted in patients with risk factors. PMID- 8717294 TI - [A high concentration of itraconazole in an aspergilloma]. AB - A 66-year-old woman was given 100 mg/day of itraconazole for eight months to treat pulmonary aspergillosis with an aspergilloma. The amount of purulent sputum decreased, but open drainage was done because the fever continued and because no improvement was seen on the chest X-ray film. No aspergillus was cultured from the surgical specimen, but fungi were observed microscopically. The concentration of itraconazole in plasma was 249 ng/ml. The concentrations in specimens of the lung and of the aspergilloma obtained by thoracotomy were 81 ng/g and 837 ng/g, respectively. The high concentration in the aspergilloma had made the fungus inactive. The itraconazole concentration may have been very high for three reasons: (1) the concentration increased in purulent fluid, (2) itraconazole easily entered the aspergilloma through the root at the cavity wall, and (3) itraconazole dissolved in lipid derived from destroyed fungus. PMID- 8717295 TI - [Intrathoracic extramedullary hematopoiesis in a case of hereditary spherocytosis]. AB - A 62-year-old man with a history of hereditary spherocytosis had an abnormal shadow on a chest X-ray film, but the shadow was not examined further. The patient was admitted to our hospital because of severe anemia, multiple gallstones, and splenomegaly. Acute cholecystitis developed due to gallstones. A smear of peripheral blood showed spherocytosis, and the osmotic fragility as measured by Parpart's method was abnormally high. These findings are consistent with hereditary spherocytosis. Splenectomy and cholecystectomy were done. The chest X-ray film, CT scan, and MRI revealed multiple well-demarcatd paravertebral masses. A biopsy of a mediastinal mass was done with an ultrasonically guided needle, and hyperplasic erythroid hemopoietic tissue was obtained. This finding led to the diagnosis of extramedullary hematopoeisis. We think extramedullary hematopoeisis should be included in the differential diagnosis of posterior mediastinal masses. PMID- 8717296 TI - [Squamous cell carcinoma of the lung associated with palmo-plantar hyperkeratosis]. AB - Palmo-plantar hyperkeratosis (PPH) is a diffuse keratoderma of the palms and soles that has been reported to be associated with internal malignancies such as lung and esophageal carcinomas. When PPH is associated with malignancy the prognosis is poor. The present patient was a 48-year-old man with PPH who suffered from squamous cell carcinoma of the lung. The pathological stage was T4N2M1 and the patient died of recurrent tumor 17 months after surgery. Because the prognosis of lung carcinoma associated with PPH is poor, patients with these conditions should undergo frequent medical examinations, which should include chest radiography and cytologic examination of sputum. PMID- 8717297 TI - [Meigs syndrome presenting as diaphragmatic pleural nodules]. AB - A 52-year-old woman was admitted to our hospital because of non-productive coughing and dyspnea on exertion. A chest X-ray film revealed a moderate pleural effusion. The effusion was persistent and progressive in spite of repeated thoracentesis. Because examinations of the effusion did not result in a specific diagnosis, thoracoscopy was done under local anesthesia. Two nodules were found in the surface of the diaphragmatic pleura. Examination of biopsy specimens revealed mesothelial hyperplasia and lymphatic duct proliferation without malignant or granulomatous lesions. Systemic examinations revealed a tumor in the right ovary. The tumor was removed, and examination revealed that it was a serous adenofibroma without malignant lesions. Meigs syndrome was diagnosed. The pleural effusion disappeared quickly, two weeks after the operation. We know of no previous report of diaphragmatic pleural nodules in a case of Meigs' syndrome. Meigs' syndrome should be considered in the differential diagnosis of pleural nodules. PMID- 8717298 TI - [Pancoast syndrome in a patient with malignant lymphoma arising from the pleura]. AB - A 79-year-old man was admitted to the hospital because of left shoulder pain. He presented with signs and symptoms typical of Pancoast syndrome. A chest roentgenogram and computed tomogram revealed a mass lesion that was connected to old pleural thicking due to tuberculosis in the left superior sulcus. A cervical lymph node biopsy was done and non-Hodgkin's lymphoma was diagnosed. Chemotherapy was effective but the patient died of severe pneumonia. Pancoast syndrome associated with malignant lymphoma is very rare. Care must be taken in the management of patients with shoulder pain, chest pain, and old pleural tuberculosis. PMID- 8717299 TI - [Diffuse tracheobronchial amyloidosis with airflow limitation and repeated pneumonias]. AB - A 67-year-old man was first admitted to our hospital complaining of a productive cough, and repeated episodes of pneumonia in different sites. Physical examination revealed expiratory wheezing and airflow limitation wlas documented with lung function tests. A chest X-ray film showed patchy pneumonic infiltrates. Chest computed tomography revealed a marked thickening of the tracheal and bronchial wall with linear calcification. Fiberoptic bronchoscopy revealed a diffuse infiltrative process in the tracheobronchial tree, which uniformly reduced the bronchial lumen. Bronchial biopsy specimens showed amyloid deposits with focal calcification, which was confirmed by Congo red and Dylon staining. Extensive examinations including rectal biopsy were negatie for systemic amyloidosis. We suggest that diffuse tracheobronchial amyloidosis should be included in the differential diagnosis of repeated pulmonary infections with airflow obstruction. PMID- 8717300 TI - [Moderately differentiated adenocarcinoma of the lung presenting as multiple intrapulmonary metastases with thin-walled cavities]. AB - A 63-year-old woman complaining of a non-productive cough was referred to our hospital. A chest X-ray film and computed tomographic scan showed a large mass in the S9 region of the right lung, and many intrapulmonary nodules with thin-walled cavities. A transbronchial biopsy specimen revealed moderately differentiated adenocarcinoma of bronchial gland origin. By the end of four cycles of chemotherapy with vindesine, ifosfamide, and cisplatin, the primary mass had markedly regressed, the many metastases had disappeared, and a few bullous lesions remained. On the second admission, many intrapulmonary metastases and cavities were seen again. Although some of the cavities may have been associated with regrowth of residual cancer cells around the remaining bullae, some nodules showed newly-developed thin-walled cavities, and in others bullous lesions developed again. These observations indicate that a check valve mechanism may operate in the formation of the thin-walled cavities. PMID- 8717301 TI - [Interstitial pneumonia associated with mixed connective tissue disease--marked improvement with azathioprine]. AB - A 55-year-old man who complained dyspnea on exertion was given a diagnosis of interstitial pneumonia associated with mixed connective tissue disease, based on symptoms and on physical and laboratory findings. Soon afterward, he experienced an acute exacerbation of the interstitial pneumonia. He was given three courses of steroid pulse therapy and cyclophosphamide, but his condition worsened. After administration of azathioprine in addition to corticosteroids, his respiratory status improved. The dose of corticosteroids was tapered and the patient has been free of disease for two years. Pulmonary involvement in mixed connective tissue disease has been considered to be relatively benign and easily treated with corticosteroids. Recently, however, many cases of corticosteroid-resistant disease with poor outcome have been reported. Azathioprine was very effective against the interstitial pneumonia in this case, but general conclusions about the most effective type of immunosuppressive agent and the timing of its administration must await the results of further study. PMID- 8717302 TI - [Acute pulmonary disease in systemic lupus erythematosus successfully treated with prednisolone pulse therapy]. AB - A 46-year-old man was admitted to the hospital because of chest pain and left pleural effusion. Fever and dyspnea developed on the third hospital day. Interstitial shadows and pleural effusions in both lower lung fields appeared on chest roentgenograms. Microscopic examination of transbronchial lung biopsy specimens taken on hospital day 5 showed thickening of the alveolar walls and desquamation of macrophages into the alveolar spaces. Analysis of bronchoalveolar lavage fluid revealed many cells and macrophages. Tests for anti-nuclear antibody and anti-DNA antibody were positive, which, in addition to serositis and proteinuria, established the diagnosis of systemic lupus erythematosus. The interstitial shadow on chest roentgenograms was believed to have reflected an acute pulmonary manifestation of systemic lupus erythematosus. The symptoms and the abnormality on chest roentgenograms were relieved within 1 month of the start of pulse therapy with prednisolone. PMID- 8717303 TI - [Pryce type I intralobar pulmonary sequestration]. AB - A 53-year-old woman had a abnormal shadow in the left lower lung field on a chest radiograph, and occasional bloody sputum. CT scans and an MRI showed an abnormal vessel shadow between the aorta and the heart. An aortogram showed an abnormal artery originating from the descending aorta and entering the left lower lobe, and a pulmonary angiogram revealed no blood supply to the left basal segments. Pryce type I pulmonary sequestration was diagnosed. The patient underwent surgery, and the abnormal artery, 13 mm in diameter, was found to be connected from the descending aorta to the left lower lobe. The abnormal artery was dissected, and the left lower lobe was removed. The postoperative course was uneventful. We collected data on 24 cases of Pryce type I pulmonary sequestration reported in Japan. Such cases are comparatively rare and males are affected more often than females. The patient in the present case was the oldest woman to have undergone surgery for this condition in Japan. In patients who underwent surgery the greatest diameter of the abnormal artery was 18 mm. PMID- 8717304 TI - [Nodular and cavitary pneumocystosis in a patient with hemophilia A and the acquired immunodeficiency syndrome]. AB - A 25-year-old man with hemophilia A who had received concentrated plasma and plasma factor VIII products since childhood presented with a productive cough and a fever. The CD4/CD8 ratio of peripheral lymphocyte subsets was very low and the serum was positive for anti-HIV antibodies. The chest roentgenogram showed bilateral multiple nodules with cavity formation. The patient underwent transbronchial lung biopsy, and the specimen obtained had Pneumocystic carinii organisms. The patient was treated with sulfamethoxazole and trimethoprim compounds, and with inhaled pentamizine. His condition improved. Patients with pulmonary pneumocystosis usually present with interstitial infiltrates spreading from the hilium to the periphery of the lungs, and nodular and cavitary lesions are unusual pulmonary radiographic findings in this condition. PMID- 8717305 TI - [Ocular metastasis necessitating enucleation of an eyeball after surgery for primary lung cancer]. AB - A 68-year-old man was referred because of an abnormal shadow in the right upper lung field on a chest X-ray film. After the diagnosis of adenocarcinoma of the lung, left upper lobectomy was done. Histologically, adenocarcinoma with foci of squamous cell carcinoma was seen. About 11 months after that operation, he suddenly noticed a decrease in visual acuity on the left side. The fundus of the eye was tested and metastatic choroidal cancer was suspected. To prevent rupture, the eyeball was enucleated. The choroidal tumor appeared to have components of squamous cell carcinoma from primary lung cancer. About 7 months after enucleation of his eyeball, the patient was disease-free with no decrease in his quality of life. As a result of a recent increase in the incidence of lung cancer, it is important to pay attention to ophthalmological tests in patients with primary lung cancer. PMID- 8717306 TI - [Acute lupus pneumonitis presenting as multiple nodular shadows]. AB - A 24-year-old woman consulted a physician because of fever, dry cough, and arthralgia of one month duration. She was transferred to our hospital for more detailed examination of pancytopenia and diffuse infiltrative shadows on a chest X-ray film. On admission she was depressed, and had oral ulcers. Pleural and pericardial effusions were observed. Laboratory studies were positive for anti nuclear antibody, anti-DNA antibody, and lupus erethymatosus. Casts were found in the urine, and systemic lypus erythematosus was diagnosed. Chest radiography and CT scan showed nodular shadows in the right S8 (2nodules with 1 cm diameter)(1 cm x 2), right S10 (0.5 cm diameter), and left S8 (0.5 cm diameter) regions, as well as diffuse infiltrative shadows in both lower lung fields. A specimen obtained by transbronchial lung biopsy revealed acute interstitial pneumonia with arteritis. After treatment with prednisolone (60 mg/day), the nodular and infiltrative shadows rapidly disappeared. Cases of lupus pneumonitis presenting as nodular shadows are very rare and are valuable in diagnostic imaging. PMID- 8717307 TI - [Tyrosine phosphorylation of focal adhesion kinase (p125FAK) and paxillin in glomeruli from diabetic rats]. AB - Focal adhesion kinase (p125FAK) is a novel non-receptor cytosolic tyrosine kinase which is activated through the phosphorylation of its tyrosine residue by ligands that bind to integrins and ligands that activate protein kinase C (PKC). In diabetic glomeruli, extracellular matrix proteins such as fibronectin, laminin and type IV collagen, which bind to integrins, were found to be increased in the mesangial area. Furthermore, PKC was shown to be activated in diabetic glomeruli. These changes might be able to cause the activation of p125FAK in diabetic glomeruli. To test this hypothesis, we examined tyrosine phosphorylation of p125FAK and paxillin, a proposed substrate of p125FAK, in glomeruli isolated from streptozotocin (STZ)-induced diabetic rats. Tyrosine phosphorylation of p125FAK or paxillin was evaluated by immunoblot analysis using anti-phosphotyrosine antibody after immunoprecipitation with anti-p125FAK or anti-paxillin antibody. Three and seven weeks after STZ injection, tyrosine phosphorylation of both p125FAK and paxillin was increased in diabetic glomeruli. The increase in tyrosine phosphorylation of p125FAK and paxillin was not observed in glomeruli from diabetic rats treated with insulin. To investigate the mechanism of increase in tyrosine phosphorylation of p125FAK, we examined tyrosine phosphorylation of p125FAK in mesangial cells plated on a fibronectin-coated dish or cultured under conditions of high glucose concentration (conditions under which PKC can be activated). Attachment of the cells to fibronectin induced tyrosine phosphorylation of p125FAK, while a high glucose concentration did not modulate tyrosine phosphorylation of p125FAK. In conclusion, tyrosine phosphorylation of p125FAK and paxillin was increased in diabetic glomeruli and these alternations may have been caused by changes in extracellular matrix proteins in diabetes. PMID- 8717308 TI - [An experimental study on the pathogenetic role of acquired resistance to acute renal failure--Enzymochemical investigation]. AB - It is an established fact that animals recovering from prior acute renal failure (ARF) are resistant to subsequent renal failure challenge with the same toxic agents, although the detailed mechanisms responsible for this phenomenon remain unclear. In this study, the mechanism underlying acquired resistance to gentanmicin (GM) was investigated from the viewpoint of kidney tissue enzymology. Sprague-Dawley rats (N = 40) were administered GM subcutaneously at the dose of 80mg/day consecutively for 40 days. Blood urea nitrogen (BUN) reached the maximum mean concentration of 36 mg/dl on day 14. Thereafter, it decreased to a level within the normal range on day 21. The change in fractional excretion of sodium (FENa) showed a curve virtually identical to the change in BUN. In renal tissue, the elevation of malondialdehyde (MDA) levels was transient during continued administration of GM. The shingomyelin (SPH)/phosphatidylcholine (PC) ratio significantly decreased on day 4, but there was no marked change thereafter. The levels of total phospholipids (PLs), phosphatidylcholine (PC), and phosphatidylethanolamine (PE) increased, whereas SPH decreased mostly on day 4. The levels of phosphatidylinositol (PI) showed a continued fall during the 40 days of the experiment. On day 40, these changes in composition recovered. Phospholipase A2 (PLA2) activities decreased gradually, whereas a distinct increase in phospholipase C (PLC) activity was maintained after day 21. Furthermore, glutathione (GSH) levels also showed two distinct cycles of decrease and increase. PLs levels correlated well with PLC activities. It was concluded that accelerated lipid peroxidation occurs early in the course of GM administration and enhances changes in the phospholipid composition, which has an influence on membrane fluidity. Thus, acquired resistance to ARF induced by GM may be due to the supply of GSH and the maintenance of alteration in phospholipid composition, which are induced by PLC activities. PMID- 8717310 TI - [Clinicopathological study of interstitial foam cells in idiopathic membranous nephropathy. Consideration of the appearance of interstitial foam cells in renal tissue]. AB - We conducted an immunohistological investigation on the pathogenesis of interstitial foam cell formation in patients with idiopathic membranous nephropathy (MN). The patients were divided into two groups: Group I consisted of 23 MN patients with interstitial foam cells; Group II consisted of the other 159 patients without foam cells. Age at renal biopsy, duration of proteinuria, blood pressure and other clinical parameters were not significantly different between the two groups. The proportion of nephrotic patients in Group I was 52.2% (12/23), and was not significantly different from that in Group II (48.4%, 77/159). Renal biopsy specimens were examined by immunoperoxidase studies using monoclonal antibodies. The interstitial foam cells were positive for EBM11 (CD68) and 25F9, which are markers of macrophage (M phi) and mature M phi, respectively, but did not express markers of T cells. In interstitial infiltrating cells, both M phi and T cells were observed, but mature M phi were seldom seen. Furthermore, LFA-1 and ICAM-1, but not ICAM-3 (the third ligand for LFA-1) were observed in the interstitial foam cells. LFA-1 and ICAM-3 were observed mainly in interstitial infiltrating cells, but ICAM-1 was observed to a much lesser extent in these cells. These results suggest that interstitial foam cells in MN may be independent of severe hyperlipidemia and proteinuria, and that there may be different mechanisms underlying the accumulation of interstitial foam cells and infiltrating m phi s. Further investigations are required to clarify the pathogenesis of interstitial foam cells in renal tissue. PMID- 8717309 TI - [Study on a predictive marker for evaluating the effectiveness of steroid therapy in IgA nephropathy-significance of extracapillary changes]. AB - In order to obtain a useful index that can predict the effectiveness of steroid therapy in IgA nephropathy, we investigated the relationship between steroid therapy and extracapillary change. We analysed 51 cases consisting of a group of 24 cases in which steroid was administered (initial dose: prednisolone 20-60 mg/day orally, or pulse therapy), and a group of 27 cases in which steroid was not administered. First of all, we compared these two groups in terms of a clinical improvement rate over 3, 12 and 60-month-periods after the therapy, respectively. It was found that steroid therapy led to a higher improvement rate (p < 0.01-0.05), indicating that steroid is effective for IgA nephropathy. Among the group undergoing steroid therapy, the frequency of cellular crescents (C) was significantly higher (p < 0.05-0.01) in the cases in whom steroid was effective and that of fibrous crescents or adhesion (F) was significantly lower (p < 0.05 0.01). We then assessed C/F in each case of the steroid-administered group, and studied its relationship with the improvement rate. As a result, the improvement rate was below 50% in the group of C/F < 0.25, over 50% of the group of 0.25 < or = C/F < 0.75, and 100% of the group of 0.75 < or = C/F. Accordingly, we concluded that C/F could be used a parameter for predicting the effectiveness of steroid therapy in IgA nephropathy and for determining indications for steroid therapy. PMID- 8717311 TI - [Excellent antiproteinuric effect by LDL apheresis in a case with severe renal dysfunction due to focal segmental glomerulosclerosis]. AB - We report here a 19-year-old man with intractable nephrotic syndrome due to focal glomerulosclerosis (FGS) treated by low-density lipoprotein apheresis (LDL-A). The patient had been receiving several drugs, including steroids, cyclophosphamide, mizoribine and deoxysparguarine, for the past ten years, but the nephrotic syndrome was resistant to these drugs. Although the initial renal biopsy specimen showed minimal change-type lesions, the second biopsy specimen obtained 6 years later revealed typical FGS findings accompanied by lipid deposition (apoB) and macrophage infiltration (CD68) in the involved area. LDL apheresis was performed ten times per course using a dextran sulfate cellulose column (Liposorba LA-15) as the LDL absorber and polysulfone hollow-fibers (Sulflux) as the plasma separator, processing a total of 3,000 ml of plasma during each apheresis. After treatment the serum levels of LDL and total cholesterol decreased to 50% and 58% of their initial levels, respectively. Immediately after the first course of treatment, the renal dysfunction did not improve, but a decrease in urinary protein was observed (from 43.7 g/day to 8 g/day). Two months later, because urinary protein increased and renal function decreased (Ccr 7 ml/min), a second course of treatment was started. However, his renal dysfunction did not improve and urinary protein did not decrease. In conclusion, in FGS with-progressive renal failure, renal histological findings of positive APO-B, CD68 (macrophage) in sclerotic lesion may be indications of effective LDL-apheresis. PMID- 8717313 TI - [A patient with Goodpasture's syndrome revealed a poor prognosis regardless of receiving intensive therapy from an early stage after onset]. AB - Goodpasture's syndrome has been reported as a disease that has a favorable prognosis when the patient receives intensive immunosuppressive drug-therapy from an early stage after onset. The present report describes a 50-year-old woman, who exhibited progressive renal failure accompanied by pulmonary hemorrhage, and an increase in serum level of antiglomerular basement membrane (GBM) antibody. Initial histological examination of a renal specimen indicated a severe fibrocellular crescentic glomerulonephritis with a linear deposition of Ig-G and C3 along the glomerular capillary wall. The patient was thus diagnosed as having Goodpasture's syndrome. Therapy with pulse treatment of steroid (corticosteroid hormone), immunosuppressive agents, or plasma-exchange for the removal of anti GBM antibody was adopted a week after the clinical onset. However, histological amelioration of the glomeruli did not occur with this treatment in the second biopsy, while glomerular damage advanced progressively. In contrast to other patients with Goodpasture's syndrome, our case revealed an unfavorable outcome regardless of receiving intensive therapy from an early period after onset, which suggests that more intensive therapy of another approach to this patient may have been necessary. PMID- 8717314 TI - [A case of mitochondrial encephalomyopathy (MELAS)]. AB - Mitochondrial encephalomyopathy is a hereditary syndrome showing impairment of muscle and the central nervous system. In this disorder, the following three syndromes have been identified on the basis of characteristic symptoms: Kearns Sayre syndrome (KSS), mitochondrial encephalomyopathy with lactic acidosis, and stroke-like episodes (MELAS), and myotonic epilepsy with ragged-red fibers (MERRF). In this report, we describe a case of mitochondrial encephalomyopathy with renal disease. A 25-year-old man was referred to our hospital in May, 1992 for evaluation of long-standing proteinuria. He had a small stature, exotropia and no pretibial edema. No mental retardation was observed. Urinary protein excretion was 2.0 g/day and urine sugar was negative. Laboratory examination revealed a serum urea nitrogen 19 mg/dl, and a creatinine value of 1.5 mg/dl. Creatinine clearance was 45.8 ml/min. His serum and spinal fluid lactate value were elevated. Biopsied muscle showed an absence of ragged-red fibers, and the presence of an A-to-G point mutation at nucleotide pari 3243 in the mitochondrial tRNA(Leu(UUR)) in peripheral blood leucocytes. He was thought to have MELAS. On the renal biopsy specimens, light microscopic examinations showed minor glomerular abnormalities with two glomerular collapses and tubulo-interstitial damage. Electron microscopic examinations showed partial thickening of the glomerular basement membrane. We report here this rare case of MELAS with renal disease, and also review seventeen cases of mitochondrial encephalopathy associated with renal disease. The existence of a relationship between mitochondrial disorder and renal damage remains obscure. PMID- 8717312 TI - [A case of hemolytic uremic syndrome accompanied by massive proteinuria and ascites]. AB - A 50-year-old male was admitted to our hospital because of proteinuria, thrombocytopenia and moderate renal dysfunction. On admission, he had massive ascites, which was transudatory. During his clinical course, renal function deteriorated and the urine volume was decreased. Hemolytic uremic syndrome with massive ascites was diagnosed based on the finding of thrombocytopenia, acute renal failure and hemolytic anemia. Methylpredonisolone pulse therapy was not effective, but plasma exchange given 22 times in total combined with vincristine sulfate, PGI2 analogue and vitamin E administration was very effective for thrombocytopenia, renal dysfunction and hemolytic anemia. Massive ascites disappeared at the same time. After complete recovery of renal function, renal biopsy was performed, revealing the reticulation of mesangial matrix and mesangiolysis, which correspond to hemolytic uremic syndrome. PMID- 8717315 TI - Epidemiological studies of radio-frequency radiation: current status and areas of concern. PMID- 8717316 TI - Cancer morbidity in subjects occupationally exposed to high frequency (radiofrequency and microwave) electromagnetic radiation. AB - Cancer morbidity was registered in the whole population of military career personnel in Poland during a period of 15 years (1971-1985). Subjects exposed occupationally to radiofrequencies (RF) and microwaves (MW) were selected from the population on the basis of their service records and documented exposures at service posts. The population size varied slightly from year to year with a mean count of about 128,000 persons each year; each year about 3700 of them (2.98%) were considered as occupationally exposed to RF/MW. All subjects (exposed and non exposed to RF/MW) were divided into age groups (20-29, 30-39, 40-49 and 50-59). All newly registered cases of cancer were divided into 12 types based on localisation of the malignancy; for neoplasms of the haemopoietic system and lymphatic organs an additional analysis based on diagnosis was performed. Morbidity rates (per 100,000 subjects annually) were calculated for all of the above localisations and types of malignancies both for the whole population and for the age groups. The mean value of 15 annual rates during 1971-1985 represented the respective morbidity rate for the whole period. Morbidity rates in the non-exposed groups of personnel were used as 'expected' (E) rates for the exposed subjects, while the real morbidity rates counted in the RF/MW-exposed personnel served as 'observed' (O) rates. This allowed the calculation of the observed/expected ratio (OER) representing the odds ratio for the exposed groups. The cancer morbidity rate for RF/MW-exposed personnel for all age groups (20-59 years) reached 119.1 per 100,000 annually (57.6 in non-exposed) with an OER of 2.07, significant at P < 0.05. The difference between observed and expected values results from higher morbidity rates due to neoplasms of the alimentary tract (OER = 3.19-3.24), brain tumours (OER = 1.91) and malignancies of the haemopoietic system and lymphatic organs (OER = 6.31). Among malignancies of the haemopoietic/lymphatic systems, the largest differences in morbidity rates between exposed and non-exposed personnel were found for chronic myelocytic leukaemia (OER = 13.9), acute myeloblastic leukaemia (OER = 8.62) and non-Hodgkin lymphomas (OER = 5.82). PMID- 8717317 TI - The influence of 50-Hz magnetic fields on cytoplasmic Ca2+ oscillations in human leukemia T-cells. AB - We have studied the effects of 50-Hz 100-microT rms magnetic fields on intracellular Ca2+ concentration in the Jurkat T lymphocyte variant E6.1 using fluorescent probes Indo-1 and Fura-2. We found, however, that the pattern of intracellular Ca2+ fluctuations also depended on the agent used for cell attachment, in our case the polypeptide poly-L-lysine. In order to isolate possible effects of magnetic field exposure from those of poly-L-lysine, the action of polypeptide on cytosolic Ca2+ was studied as well. It was found that a 10(-7)% concentration of polypeptide triggered prolonged Ca2+ spiking. Higher (10(-4)%) concentrations induced rapid increases in intracellular Ca2+ followed by high, unstable Ca2+ levels. The response of these cells to the monoclonal antibody anti-CD3 was also inhomogeneous, similar to one caused by poly-L-lysine. The effect of magnetic field exposure was studied on cells initially exhibiting (1) non-oscillating, low Ca2+ concentration and (2) prolonged Ca2+ concentration oscillations. In case (1) the result was negative. In case (2), statistically significant changes were found: the oscillation amplitude was reduced on average by 30%, and the frequency composition was shifted towards higher frequencies. PMID- 8717318 TI - Experimental study of the effects of radiofrequency electromagnetic fields on animals with soft tissue wounds. AB - The effect of radio frequency electromagnetic fields (RF EMF) was studied on Wistar rats with excised full-thickness dermal wounds in the interscapular region. The wounded regions of experimental animals were subjected to EMF for 30 min daily during the first 5 days after wound infliction. Control animals received no treatment. We used RF EMF with (1) frequency 53.53 GHz without modulation; (2) frequency 42.19 GHz without modulation; (3) frequency 42.19 GHz, but with a frequency modulation band 200-MHz wide. On the 7th day the animals were terminated and the granulation-fibrous tissue (GFT) developed in the wounds was subjected to complex quantitative biochemical analysis. RF EMF without frequency modulation decreased the amounts of glycoprotein macromolecules, diminishing the inflammatory exudation. In striking contrast, under the influence of RF EMF with frequency modulation, hexoses and especially sialic acid concentrations were significantly elevated (P < 0.001). This indicated intensification of exudative phenomena. As a consequence of inflammation inhibition in the treatment without frequency modulation, the total collagen accumulation was lowered. However, when frequency was modulated, the inflammatory phenomena were intensified, and pronounced accumulation of collagenous proteins was noted. Thus, our experiments confirm the effects of non-thermal EMF on the reparative-proliferative processes of animals with soft tissue wounds. PMID- 8717319 TI - Assessment of radio-frequency electromagnetic radiation by the micronucleus test in bovine peripheral erythrocytes. AB - Previous bioindicative studies in the Skrunda Radio Location Station area have focused on the somatic influence of electromagnetic radiation on plants, but it is also important to study genetic effects. We have chosen cows as test animals for cytogenetical evaluation because they live in the same general exposure area as humans, are confined to specific locations and are chronically exposed to radiation. Blood samples were obtained from female Latvian Brown cows from a farm close to and in front of the Skrunda Radar and from cows in a control area. A simplified alternative to the Schiff method of DNA staining for identification of micronuclei in peripheral erythrocytes was applied. Microscopically, micronuclei in peripheral blood erythrocytes were round in shape and exhibited a strong red colour. They are easily detectable as the only coloured bodies in the uncoloured erythrocytes. From each individual animal 2000 erythrocytes were examined at a magnification of x 1000 for the presence of micronuclei. The counting of micronuclei in peripheral erythrocytes gave low average incidences, 0.6 per 1000 in the exposed group and 0.1 per 1000 in the control, but statistically significant (P < 0.01) differences were found in the frequency distribution between the control and exposed groups. PMID- 8717320 TI - Motor and psychological functions of school children living in the area of the Skrunda Radio Location Station in Latvia. AB - This paper presents the results of experiments on school children living in the area of the Skrunda Radio Location Station (RLS) in Latvia. Motor function, memory and attention significantly differed between the exposed and control groups. Children living in front of the RLS had less developed memory and attention, their reaction time was slower and their neuromuscular apparatus endurance was decreased. PMID- 8717321 TI - [Prof. Dr. of medicine Johannes Staudt 1932-1994]. PMID- 8717322 TI - Simple or repeated induction of superovulation: a study on ovulation rates and microvessel corrosion casts in ovaries of golden hamsters. AB - Repeatedly stimulated ovaries are reported to decrease the ovulation rate. One cause among others might be that the microvascular bed has been insufficiently developed. Therefore, 30-day-old golden hamsters were superovulated either once or repeatedly. At the light microscopic level, the ovulation rate in serially sectioned ovaries was indirectly determined by the occurrence of corpora lutea and of abnormal follicle rupture with oocyte release into the cortical stroma (IOR). For the study with scanning electron microscopy (SEM), the microvascular bed of the ovaries was cast with a polyester resin, and the corrosion casts of mature follicles observed. The histological sections of once-stimulated ovaries showed a large number of corpora lutea and IOR follicles. This indicated hyperovulation. In corrosion casts of once-stimulated ovaries, large-sized antral follicles with two layers of a dense capillary meshwork were observed. Capillary sprouts were aligned around the antrum 0 to 12 h after administration of human chorionic gonadotrophin (hCG), and these radiated towards the center of the antrum after 12 to 36 h had elapsed. The ovulation site was recognized at the follicle apex by three similarly sized structures which were either a sinusoid, an oocyte replica, or an opening. Repeatedly stimulated ovaries produced a low number of corpora lutea and almost no IOR follicles. This was judged as hypoovulation. The microvessels of mature follicles were reduced in number and incompletely cast. Widespread resin leakages were conspicuous in the follicle wall 36 h after hCG injection, but the capillary sprouts radiated towards the center of the antrum. No ovulation site was detectable. It is concluded, that capillary sprouts are induced before luteinization. The ovulation site is indicated by particular changes in its microvascular bed. An insufficiently developed microvascular bed may be responsible for hypoovulation in repeatedly stimulated ovaries. PMID- 8717323 TI - Development of the motor endplates in the masseter muscle in the human fetus. AB - The development of the motor endplate (MEP) and the structure of the masseter muscle in the human fetus were examined by light and electron microscopy. At 12 weeks of gestation, the masseter muscle was composed mostly of irregularly arranged myotubes. The number of muscle fibers increased while that of the myotubes decreases during development. After 28 weeks of gestation, the masseter muscle was composed only of muscle fibers, and these fibers rapidly increased in size in comparison with those prior to 28 weeks of gestation. The MEP appeared during the first 12 weeks of gestation, and were of an undeveloped simple type, with only one axon branch. After 20 weeks of gestation, MEPs were classified as complex or simple types in terms of the branching of the axons. The complex type of MEP was found in muscle fibers of large diameter, while the simple type was found in muscle fibers of small diameter. Schwann-like cells appeared at the surface of the MEP. After 28 weeks of gestation, only the complex type was found in the masseter muscle. These results suggest that, the development of the MEP is closely related to the development of muscle, and that the timing of the development of the masticatory muscles differs from that of other skeletal muscles, such as those in the trunk and limb. PMID- 8717324 TI - Demonstration of amoeboid and ramified microglial cells in pre- and postnatal bovine brains by lectin histochemistry. AB - In embryonic, fetal and postnatal bovine brains the development and distribution of microglial cells was examined by lectin histochemistry, using the isolectin B4 from Griffonia simplicifolia (GSA I-B4), the lectin from Ricinus communis (RCA I), and mistletoe lectin (ML I). With GSA I-B4 and ML I, different types of microglial cells, i.e., amoeboid, intermediate and ramified cells, were specifically stained. On sections fixed in Bouin's fluid significantly higher numbers of microglial cells were labelled than on sections fixed in formalin. On the latter, proteolytic pretreatment was required. With RCA-I, no staining of microglial cells was achieved. This finding may indicate the presence of very low concentrations of beta-D-galactose residues on bovine microglial cells in comparison with other species studied so far. In the fetal telencephalon, the highest numbers of amoeboid microglial cells were found in transitory structures (subependymal regions of the lateral ventricles, cavum septi pellucidi, intermediate zone) and in areas of developing axon tracts (corpus callosum, internal and external capsules) between three and five months of gestational age. From 3-4 months of gestational age onward, the appearance of ramified microglial cells was noted. In 7-8 month-old fetuses, a complete change of the microglial cell picture occurred. Ramified cells clearly predominated, whereas amoeboid cells had markedly decreased. In 8-9 month-old fetuses, amoeboid microglial cells had almost disappeared from fetal brains. In brains from subadult and adult cattle, lectin-positive ramified microglial cells with up to five cellular processes were seen in all brain areas, located adjacent to vessels or surrounding neuronal perikarya. PMID- 8717325 TI - The expression of VLA integrins in the human thymus. AB - The integrin receptors are a family of transmembrane glycoproteins comprising non covalent heterodimers. They interact with a wide variety of ligands including extracellular matrix glycoproteins, complement and other cells while their intracellular domains interact with the cytoskeleton. They participate in cell matrix and cell-cell adhesion in many physiologically important processes including embryological development, hemostasis, thrombosis, wound healing, immune and nonimmune defense mechanisms, and oncogenic transformation. This investigation is focused on the histological distribution of the beta 1-integrins in the human thymus, using an indirect immunoperoxidase method. With the exception of VLA-4, none of the beta 1 integrins were expressed on thymocytes which were strongly positive in the cortex and perivascular compartment, somewhat weaker in the medulla. Thymic epithelial cells were positive for VLA-1, VLA-2, VLA-3 and VLA-6, but the distribution pattern of these molecules in epithelial cells at certain locations was quite different. VLA-1 was weakly expressed by both cortical and medullary epithelial cells. VLA-2 was strongly positive in cortical epithelial cells forming a dense framework at the peripheral cortex. VLA 3 and VLA-6 selectively stained a single flattened epithelial cell layer (perilobular epithelial cells) demarcating the peripheral cortex from the surrounding perivascular compartment. VLA-1,3,5,6 were also demonstrated in the endothelial cells and subendothelial layer of the thymic vasculature. IN CONCLUSION: the distribution of integrins in human thymus tissues is of special interest. Such distribution shows that the VLA integrins may have different functions in different areas. The data presented in this study may be important in evaluating the functional role of the VLA integrins in thymocyte maturation in different compartments of the thymus. PMID- 8717327 TI - Morphology of the antebrachial musculature of the American kestrel, Falco sparverius (aves). AB - The antebrachial musculature of the American kestrel (Falco sparverius) is described. This fills a gap in the avian morphology literature, and provides a reference for future comparative, functional and systematic studies. A table of synonyms-homologs is provided for each muscle as a reference frame for over 100 years of avian anatomical literature. PMID- 8717326 TI - Morphological features and kinetics of primary cultures of BPH tissue supported by TV1 medium. AB - 222 human prostatic biopsies were used to prepare cell cultures by means of a medium--colony formation permissive--containing fetal calf serum, called TV1. After 7, 14 and 21 days, the cultures were examined by optical and scanning electron microscopy. TV1 medium induces the formation and growth of two types of colonies, one mainly composed of epithelioid cells and distinguished by early growth; the second one made up exclusively of fibroblastoid cells which appear later in the culture. Epithelioid colonies, comprising three different cell types, appear to be arranged as a growth halo concentric to the bioptic fragment with a large central area, formed by a monolayer, and a pluristratified edge. Fibroblastoid cells weakly adhere to the substrate and form "satellite growth halos" separated from the primitive bioptic fragment. All the epithelioid cells were positive to cytokeratin LP34 Mab and negative to anti-smooth muscle-actin and anti-proline-4-hydroxylase antibodies. Fibroblastoid cells were only anti proline-4-hydroxylase positive. The cell kinetics of epithelioid cells were also studied, revealing an extension of the S phase, in contrast to what happened with WAJC 404, and consequently a reduction of the percentage of cells entering mitosis. For this reason, the addition of fetal serum to the culture medium does not allow the use of prostate primary cultures for more than 14 days. PMID- 8717328 TI - Bilateral duplication of the optic canals. AB - During an autopsy survey of 369 human skulls, one with bilateral (0.27%) and one with unilateral (0.27%) duplication of the optic canal were reported. The anatomical findings in case of this rare anomaly are presented. PMID- 8717329 TI - Histochemical study of muscle fiber types in Synbranchus marmoratus bloch, 1795. AB - The myotomal muscle of Synbranchus marmoratus was investigated using histochemical and immunohistochemical reactions. This musculature is composed of a superficial red compartment, uniformly distributed around the trunk circumferentially and also in the lateral line. The red compartment fibers are small in diameter and have an oxidative metabolism, a high rate of glycogen and a negative reaction to alkaline and acid myofibrillar ATPase (mATPase). The white muscle forms the bulk of the muscle mass. Its fibers are large in diameter and have a glycolytic metabolism, a negative reaction to glycogen, a strong reaction to alkaline mATPase and a negative reaction to acid mATPase. Between these two compartments there is an intermediate layer of fibers presenting a mosaic metabolism pattern with a high rate of glycogen. These fibers stained moderately for alkaline and acid m-ATPase. Several clusters of red muscles were observed inside the white muscle. Each cluster is composed of three fiber types, with a predominance of red and intermediate fibers. Reactivity to anti-MHC BA-D5 was positive only in the intermediate fibers. Reactivity to anti-MHC SC-71 was negative in all fiber types. PMID- 8717330 TI - Hard tissue of teeth and their calcium and phosphate content in Ambystoma mexicanum (Urodela: Ambystomatidae). AB - The wall of the pulp cavity, fracture faces and the demineralized surfaces of teeth from larvae and adults of Ambystoma mexicanum were investigated by scanning electron microscopy (SEM). Calcium and phosphate contents were determined by microanalysis. The apical part of the tooth (crown, tooth apex) contains dentin canals. In the larva, these do not reach the enamel-dentin border but end below this border in front of a denser hard substance, possibly enameloid. The pedicel in the adult and the basal portion of the tooth in the larva (base) are without dentin canals. These parts of the teeth are characterized by longitudinally arranged collagen fibres as visualized on the demineralized surfaces. These observations indicate a congruency in early-larval and adult teeth between base and pedicel as well as apex and crown. This partition is also confirmed by the calcium and phosphate values which were identical in larvae and adults. Highest values are found in enamel and lowest values in the tooth-bearing bone. Calcium and phosphate content show a clear difference between dentin and the basal part of the tooth (pedicel and base). The ring-like dividing zone in the adult tooth is less well mineralized. PMID- 8717331 TI - Distribution pattern of HSP60 immunoreactivity in the testicular tissue of infertile men. PMID- 8717332 TI - Classification of the vascular patterns of the thoracic limb muscles of cats. AB - The vascular patterns of the thoracic limb muscles were studied in 20 limbs of 10 cats. Gross dissection and radiography were used to identify the vascular pedicles. It is proposed that the latissimus dorsi, the cervical portion of the trapezius, the caudal portion of the omotransversarius, the deep pectoral, the deltoid, the biceps brachii, the lateral digital extensor and the extensor carpi ulnaris muscles are suitable for making muscular and musculocutaneous flaps in cat. PMID- 8717333 TI - Development of the human fetal visceral pleura. An ultrastructural study. AB - The visceral pleura of human fetuses aged from 9 to 36 weeks of gestation was studied by means of transmission electron microscopy. The main components of the visceral pleura (mesothelium, basal lamina and submesothelial connective tissue layer) are formed in the fetal period. They develop asynchronously in different pleural areas, and do not reach maturity. Fetal differentiation of the lung pleura can be divided in two stages--early (until 17 gestation week) and late stage--up to birth. The high mesothelial cells appear later than the flat cells, but the first type predominates in the final covering layer during the period investigated. The most significant developmental phenomena of the mesothelium involve membranous differentiation (the microvillous covering, vesicular system and intercellular contacts). The different transport and secretory potentials of the mesothelial cells during the various prenatal periods are discussed. The mode of development of the basal lamina suggests its mesothelial origin. The elastic membrane appears during the late stage of fetal life. The components of the submesothelial connective tissue layer (fibroblasts, collagen and elastic fibres, blood and lymph vessels) undergo several phases of differentiation. PMID- 8717334 TI - Correlation between increased AP-1NGF binding activity and induction of nerve growth factor transcription by multiple signal transduction pathways in C6-2B glioma cells. AB - Transcription mechanisms regulating nerve growth factor (NGF) gene expression in the CNS are yet to be thoroughly understood. We have used C6-2B rat glioma cells to characterize the signal transduction pathways that contribute to transcriptional and posttranscriptional regulation of NGF mRNA. Because the NGF promoter contains an AP-1 consensus sequence, we have investigated whether increases in AP-1 binding activity correlate with enhanced NGF mRNA expression. Gel mobility shift assays using an oligonucleotide homologous to the AP-1 responsive element of the rat NGF gene (AP-1NGF) revealed that 12-O-tetradecanoyl phorbol-13-acetate (TPA) and, to a lesser extent, isoproterenol (ISO) and thapsigargin, a microsomal Ca(2+)-ATPase inhibitor, stimulated binding to AP-1NGF within 2 h. All of these stimuli increased NGF mRNA levels within 3 h. Cycloheximide pretreatment blocked the TPA and ISO-mediated binding to AP-1NGF suggesting that de novo synthesis of c-Fos/c-Jun may be required for the transcriptional regulation of NGF gene. Nuclear run-on assays and NGF mRNA decay studies revealed that TPA increases NGF transcription whereas ISO affects both transcription and mRNA stabilization. We propose that (i) different signal transduction mechanisms regulate the expression of the NGF gene in cells derived from the CNS, and (ii) both mRNA transcription and stability account for the cAMP mediated increase in NGF mRNA levels. PMID- 8717335 TI - Developmental expression of functional GABAA receptors containing the gamma 2 subunit in neurons derived from embryonal carcinoma (P19) cells. AB - The expression of the gamma 2 subunit into functional GABAA receptors has been examined in the embryonal carcinoma (EC) cell line P19, a pluripotent cell line which differentiates into a neuronal phenotype after exposure to retinoic acid. Whole-cell voltage-clamp recordings were used to examine the characteristics of the GABA receptors expressed in P19 cells at different times after exposure to retinoic acid. Messenger RNA for both the gamma 2L and gamma 2S splice variants of the GABAA receptor increased dramatically following differentiation of P19 EC cells with retinoic acid. By 12 days after retinoic acid treatment, while both mRNAs were present, there was an approximately 10-fold greater abundance of gamma 2S mRNA compared to gamma 2L. However, at this same time point neurons derived from P19 cells stained intensely with a polyclonal antibody raised against a peptide fragment specific for the gamma 2L subunit. A significant increase in both the affinity for GABA and the maximum current amplitude elicited by GABA occurred between 7 and 12 days after retinoic acid treatment. In contrast, the ability of the benzodiazepine agonist flurazepam to potentiate GABA-induced membrane current was the same at 7 and 12 days after retinoic acid treatment. These data suggest that the gamma 2 subunit of the GABAA receptor is expressed early following differentation of P19 cells into a neuronal phenotype, and that this subunit is incorporated into functional GABAA receptors. Moreover, the gamma 2S and gamma 2L splice variants of this subunit may be co-expressed in neurons derived from P19 cells. The observed affinity change for GABA may reflect a time dependent change in the expression of alpha and/or beta subunits of the GABAA receptor, as occurs in developing neuronal tissue both in vitro and in vivo. PMID- 8717336 TI - Gamma-radiation-induced cell death in the fetal rat brain possesses molecular characteristics of apoptosis and is associated with specific messenger RNA elevations. AB - Low-dose ionizing irradiation of 16-18-day pregnant rats rapidly kills stem cells in the fetal forebrain. We have examined gamma-irradiated 17-day fetal rat brain tissue for molecular characteristics of apoptosis and changes in levels of mRNAs relevant to apoptosis. In many forebrain cells radiation elicits within 5 h nuclear condensation and fragmentation consistent with apoptosis. An electrophoretic DNA ladder indicative of internucleosomal chromatin cleavage was prominent within 3 h after irradiation. Pretreatment of pregnant rats with cycloheximide, or pretreatment of dissociated fetal brain cells in culture with actinomycin D, abolished the radiation-induced internucleosomal DNA fragmentation, demonstrating requirements for protein and RNA synthesis. Irradiation dramatically increased the level of the p53 transcription factor and the abundances of mRNAs coding for the cell-cycle inhibitor p21/Waf-1/Cip-1 and the AP-1-associated transcription factors Fos and JunB. Irradiation moderately increased the level of mRNA for the positive apoptosis regulator Bax. In contrast, irradiation reduced by 50-70% the abundances of most other mRNAs tested, including those for housekeeping proteins, p53, Jun, Myc, interleukin-1 beta-converting enzyme, and the negative apoptosis regulators Bcl-2 and Bcl-xL. These results indicate that radiation-elicited apoptosis of fetal brain cells is associated with activation of the p53 system, probable increases in AP-1 Fos/JunB heterodimers, and an increased ratio of Bax to Bcl-2 + Bcl-xL. PMID- 8717337 TI - Molecular cloning of novel leucine-rich repeat proteins and their expression in the developing mouse nervous system. AB - It is well established that leucine-rich repeat (LRR) proteins such as connectin, slit, chaoptin, and Toll have pivotal roles in neuronal development in Drosophila as cell adhesion molecules. However, to date, little information concerning mammalian LRR proteins has been reported. In the present study, we sought LRR proteins of the mouse brain, based on the assumption that fundamental mechanisms are conserved between different species. We screened a neonatal mouse brain cDNA library with a human partial cDNA encoding LRR protein as a probe. We obtained two independent cDNAs encoding LRR proteins, designated NLRR-1 and NLRR-2 (Neuronal Leucine-Rich Repeat proteins). We analyzed the whole sequence of NLRR-1 and partial sequence of NLRR-2. Sequence analysis showed that these two clones are about 60% homologous to each other, and that NLRR-1 protein is a transmembrane protein. Northern blot analysis and in situ hybridization histochemistry showed that both NLRR-1 and NLRR-2 mRNAs were expressed primarily in the central nervous system (CNS); NLRR-1 mRNA was also detected in the non neuronal tissues such as cartilage, while NLRR-2 mRNA expression was confined to the CNS at all developmental stages. These results suggest that there is at least one LRR protein family in the mouse and that these molecules may play significant but distinct roles in neural development and in the adult nervous system. PMID- 8717338 TI - Alpha 2 noradrenoceptors in the anterior piriform cortex decline with acute amino acid deficiency. AB - The responses of the brain to the amino acid deficiency that occur after eating imbalanced amino acid diets (IMB) have been associated with decreased concentrations of norepinephrine (NE) and cAMP in the anterior piriform cortex (APC), an area essential for the initial feeding responses to amino acid deficiency. In addition, the anorectic responses to IMB were decreased after injections of the alpha 2 agonist, clonidine, and increased after injections of the alpha 2 antagonist, idazoxan, into the APC. Therefore, to study the role of the alpha 2-noradrenergic receptor further in this model, we measured alpha 2 noradrenergic receptor binding in the APC of rats fed two levels of threonine IMB or a low-protein basal control diet. After basal prefeeding for 10 days, rats were given either a mild IMB, a severe IMB, or the basal diet for 2.5 h. The APC, anterior cingulate cortex (AC), ventromedial hypothalamus (VMH), and lateral hypothalamus (LH) were assayed. Binding of [3H]p-aminoclonidine to alpha 2 receptors determined that alpha 2 binding was decreased the most in APC (P < 0.0003). Binding in APC was significantly correlated with food intake in the anorectic response to IMB (P < 0.001). In AC, binding was also significantly decreased, but less dramatically (P = 0.012), and was not correlated with food intake. There were no significant changes in LH or VMH, although alpha 2 noradrenergic binding in VMH tended to decrease with the severe IMB in a pattern similar to APC. Plasma glucose values did not differ after the same feeding protocol. These data support our hypothesis that NE activity in the APC plays a role in initiating the anorectic response to IMB, perhaps via the alpha 2 noradrenergic receptor. PMID- 8717339 TI - Developmental expression in the mouse nervous system of the p493F12 SAP kinase. AB - Mitogen-activated protein (MAP) kinases are proline-directed, serine/threonine kinases that respond to a variety of extracellular signals. A subgroup of these kinases, stress-activated protein (SAP) kinases, phosphorylate c-jun in response to cellular stress. Using monoclonal antibody (MAb) 3F12, we have cloned and partially characterized p493F12 kinase, a mouse homologue of the rat SAP beta kinase and described its expression in the adult and developing mouse. Unlike previously reported MAP and SAP kinases, it is primarily expressed as a 2.7 kb transcript in neurons in the nervous system of the adult mouse. A 2.4 kb transcript is also expressed in the testis. Immunocytochemically, MAb 3F12 decorates a loop-like structure encircling the nucleus in the cytoplasm of neurons in the adult brain, and distinct perinuclear dots in the embryos. In situ hybridization first reveals expression in post-mitotic neurons, on embryonic day 11. The mRNA is also expressed in the Neuro-2A neuroblastoma cell line and is not upregulated in response to differentiating agents. The neuronal specificity of this kinase suggests the presence of a signal transduction cascade unique to neurons. As the amino acid sequence is highly conserved in the human and mouse, the latter may serve as a model for regulation and expression of this kinase. PMID- 8717340 TI - Production and increased detection of amyloid beta protein and amyloidogenic fragments in brain microvessels, meningeal vessels and choroid plexus in Alzheimer's disease. AB - Recent advances indicate soluble amyloid beta (A beta) protein is produced constitutively during normal metabolism of the amyloid precursor protein (APP). This has not been directly examined in human brain vascular tissues. Using a panel of well-characterized antibodies, here we show that increased amounts of soluble A beta were found in isolated vascular tissues from AD subjects compared to age-matched controls without significant Alzheimer pathology. Immunocytochemical analyses of isolated vessel preparations showed characteristic transverse patterns of A beta deposits in large vessels with smooth muscle, however, fine A beta deposits were apparent even in capillaries. A proportion of such A beta protein and potentially amyloidogenic carboxyl terminal fragments were released by solubilization and disruption of the vascular basement membrane by collagenase treatments. We further demonstrated by in vitro metabolic labelling that soluble A beta or an A beta-like peptide is associated and produced by cerebral microvessels, meningeal vessels and the choroid plexus isolated postmortem from human as well as rat brain. Compared to those from young rats, cerebral microvessels from aging rats showed increased release of carboxyl terminal fragments of APP and A beta-like peptide. Our observations provide the first direct demonstration that human vascular tissues produce soluble A beta, a product of the secretory pathway in APP processing. Our findings also suggest that aging associated alterations in the basement membranes are a factor in A beta accumulation that results in vascular amyloid deposition, the principal feature of cerebral amyloid angiopathy. PMID- 8717341 TI - Amphetamine and vigabatrin down regulate aromatic L-amino acid decarboxylase mRNA levels. AB - Aromatic L-amino acid decarboxylase (AADC) has previously been shown to be up regulated at the level of its protein activity and its mRNA abundance by antipsychotic drugs. Its activity has also been shown to be down-regulated by dopamine agonists including amphetamine. In this study we have injected rats for up to 32 days with amphetamine and the anti-epileptic drug vigabatrin, both of which can cause psychosis with similarities to schizophrenia. We have shown that AADC mRNA levels are reduced in most brain regions by both drugs. Cocaine and other non-psychotogenic anti-epileptic drugs had no effect in this paradigm. Two products of this enzyme are implicated in psychotogenesis. PMID- 8717342 TI - Hippocampal somatostatin receptors and modulation of adenylyl cyclase activity in histamine-treated rats. AB - In the present study, the effects of an intracerebroventricular (i.c.v.) dose of histamine (0.1, 1.0 or 10.0 micrograms) on the hippocampal somatostatin (SS) receptor/effector system in Wistar rats were investigated. In view of the rapid onset of histamine action, the effects of histamine on the somatostatinergic system were studied 2 h after its administration. Hippocampal SS-like immunoreactivity (SSLI) levels were not modified by any of the histamine doses studied. SS-mediated inhibition of basal and forskolin (FK)-stimulated adenylyl cyclase (AC) activity was markedly increased in hippocampal membranes from rats treated with 10 micrograms of histamine (23% +/- 1% vs. 17% +/- 1% and 37% +/- 2% vs. 23% +/- 1%, respectively). In contrast, neither the basal nor the FK stimulated enzyme activities were affected by histamine administration. The functional activity of the hippocampal guanine-nucleotide binding inhibitory protein (Gi protein), as assessed by the capacity of the stable GTP analogue 5' guanylylimidodiphosphate (Gpp[NH]p) to inhibit FK-stimulated AC activity, was not modified by histamine administration. These data suggest that the increased response of the enzyme to SS was not related to an increased functional activity of Gi proteins. In fact, the increased AC response to SS in hippocampal membranes from histamine (10 micrograms)-treated rats was associated with quantitative changes in the SS receptors. Equilibrium binding data obtained with [125I]Tyr11 SS indicate an increase in the number with specific SS receptors (541 +/- 24 vs. 365 +/- 16 fmol/mg protein, P < 0.001) together with a decrease in their apparent affinity (0.57 +/- 0.04 vs. 0.41 +/- 0.03 nM, P < 0.05) in rat hippocampal membranes from histamine (10 micrograms)-treated rats as compared to control animals. With the aim of determining if these changes were related to histamine binding to its specific receptor sites, the histaminergic H1 and H2 receptor antagonists mepyramine and cimetidine, respectively, were administered 1 h before histamine injection. The pretreatment with mepyramine or cimetidine induced an increase in the number and affinity constant of the SS receptors whereas the simultaneous pretreatment with both histamine antagonists prevented the histamine induced changes in SS binding to its receptors. Since the hippocampal SS receptor/effector system is modulated by histamine, it is tempting to speculate that in the hippocampus, SS could be involved as a mediator of the histamine effects on behaviors such as learning and memory. PMID- 8717343 TI - Altered brain sodium channel transcript levels in human epilepsy. AB - Normal, and perhaps pathological, characteristics of neuronal excitability are related to the distribution and density of voltage-gated ion channels such as the sodium channel. We studied normal and epileptic human brain using the ligase detection reaction to measure the relative quantities of mRNAs encoding sodium channel subtypes 1 and 2. Normal brains exhibited characteristic 1:2 ratios which varied by brain region, but the ratios were invariate among individuals. These normal values were altered as much as threefold in anatomically corresponding regions of epileptic brain tissues. Changes of this magnitude in such a highly conserved value support a potential role for sodium channels in the pathophysiology of epilepsy. PMID- 8717344 TI - Altered expression of gamma 2L and gamma 2S GABAA receptor subunits in the aging rat brain. AB - Aging-related alterations in both protein and mRNA expression of gamma 2S and gamma 2L subunits of the GABAA receptors have been observed in several brain areas of Sprague-Dawley and Fischer 344 rats. Subunit-specific antibodies to gamma 2S and gamma 2L as well as a riboprobe to the large intracellular loop of gamma 2, which recognizes both gamma 2S and gamma 2L mRNAs, in conjunction with computerized image analysis were used for quantitative immunocytochemistry and in situ hybridization. In addition, specific oligonucleotide probes to gamma 2S or gamma 2L mRNA were used for quantitative dot blot hybridization. A large increase in the number of heavily immunostained neurons with the anti-gamma 2L antibody was detected in the cerebral cortex (115%) of old rats. However, only a small (but significant) aging-related increase in the density of gamma 2L immunostaining (7%) was observed throughout the cerebral cortex whereas no significant aging-related change in gamma 2L mRNA was detected in this brain region. Contrary to gamma 2L, the gamma 2S immunostaining did not show aging related increased number of heavily immunostained neurons in cerebral cortex. Moreover, the density of gamma 2S immunostaining and the expression of gamma 2S mRNA were significantly decreased in the cerebral cortex (9-24%). Important aging related changes were also found in the cerebellum of old rats where the expression of both gamma 2S and gamma 2L peptides was significantly decreased (24% and 23% respectively). This decrease in gamma 2 protein expression was accompanied by decreased expression of gamma 2S (16-38%) and gamma 2L (24%) mRNAs. Nevertheless, the most important decrease of gamma 2S (48%) and gamma 2L protein (20%) was revealed in the molecular layer of the cerebellum. In addition, the expression of gamma 2S protein was increased (14%) whereas the expression of gamma 2L was decreased (13%) in the granule cell layer. Therefore, the relative expression of gamma 2S protein in both layers was reversed in old animals. The observed aging-related changes in the expression of GABAA receptor subunits might lead to altered GABAA receptor/benzodiazepine receptor subunit composition. PMID- 8717345 TI - Regulation of somatostatin gene expression by veratridine-induced depolarization in cultured fetal cerebrocortical cells. AB - The stimulatory effect of veratridine (VTD) depolarization upon somatostatin mRNA (SS mRNA) levels in primary cultures of fetal cerebrocortical cells was analyzed. Depolarizing stimuli, such as 100 microM VTD exposure for 30 min, elicited an increase in immunoreactive somatostatin (IR-SS) release to the media without affecting SS mRNA levels. These levels increased when exposure to depolarization stimuli was prolonged up to 3 or more hours. At this time, veratridine acted as a secretagogue, stimulating somatostatin secretion, but was also effective in stimulating somatostatin mRNA levels. These changes were blunted by the Na+ channel blockade tetrodotoxin (TTX), and partially abolished by the Ca2+ channel antagonist, verapamil (VPM). To study whether VTD may affect mRNA stability we determine the rate of disappearance of SS mRNA after inhibition of transcription by actinomycin D and demonstrated that VTD stimulation did not stabilize the SS mRNA. These results indicate that the induction of SS mRNA expression by VTD involves the modulation of Ca2+ and Na+ channels. The time course study confirmed that the VTD-induced SS mRNA accumulation is time-dependent, and requires a prolonged exposure to stimulate SS gene expression. VTD stimulation does not modify the SS mRNA rate of degradation. PMID- 8717346 TI - Regulation of actin and tubulin gene expression by thyroid hormone during rat brain development. AB - In the developing brain the active neurite outgrowth during the early phase of synaptogenesis is associated with a thyroid hormone dependent expression of tubulin and actin. In this study, the molecular mechanism of thyroid hormone (TH) action on actin and tubulin gene expression in the developing rat brain has been investigated by comparing the steady state levels of both mRNAs with their respective rates of transcription in cerebra from normal and hypothyroid animals. The developmental profile of actin as well as tubulin mRNAs in both normal and hypothyroid brains display a biphasic pattern, increasing progressively during the first week after birth and declining thereafter. However, hypothyroidism resulted in a significant reduction in the steady state levels of both mRNAs during the first postnatal week. During the second and third weeks, in contrast to their rapid decline in the normal controls, the corresponding decrease in the hypothyroid cerebra was retarded and prolonged resulting in their higher levels under TH-deficient condition. Kinetics of stimulation of actin and tubulin mRNAs in the 5-day hypothyroid cerebra following injection of the optimal dose of TH (200 micrograms T3/100 g body wt.) demonstrated elevation of both mRNAs within 1 h indicating a possible role of TH at the transcriptional level. In vitro transcription experiments by nuclear run off assay unambiguously confirmed that actin gene transcription is depressed in the hypothyroid cerebra compared to normal control. This reduced rate of transcription could be significantly induced in the hypothyroid cerebra by incubation of hypothyroid nuclei with T3 prior to transcription. In contrast, except for a reduced transcription in 5-day hypothyroid nuclei, no effect on tubulin gene transcription was evident at any other age. Moreover preincubation of hypothyroid nuclei from all three ages with T3 had no stimulatory effect on tubulin gene transcription. Analysis of age related changes in the rates of transcription of actin and tubulin genes with their corresponding steady state mRNA levels in normal and hypothyroid developing brain provides strong evidence that although additional modes of regulation may be operative, transcription represents an important level of control for thyroidal regulation of actin gene expression while tubulin gene expression is primarily regulated at post-transcriptional level. PMID- 8717347 TI - Characterization of two fish glutamate receptor cDNA molecules: absence of RNA editing at the Q/R site. AB - Two cDNA clones encoding putative ionotropic glutamate receptor subunits were isolated from a brain cDNA library of a freshwater fish, Oreochromis sp. The deduced amino acid sequences of these two cDNAs, fGluR2 alpha and fGluR2 beta, display the highest sequence identity (85%) to that of the rat GluR2 (AMPA receptor subunit) and they contain an arginine codon at the Q/R editing site of the TM2 segment. Genomic sequence analysis of the exons encoding the TM2 reveals the presence of an arginine codon at the Q/R site, suggesting that the RNA editing mechanism acting in the mammalian GluR2 does not operate at the homologous site in these two fish genes. In contrast to the absence of RNA editing at the Q/R site, transcripts of fGluR2 alpha and fGluR2 beta are subjected to RNA editing at a second site, the R/G site. A splicing variant of fGluR2 alpha, fGluR2 alpha-c, with a shorter C-terminal sequence was found; however, no C-terminal splicing variant of fGluR2 beta was detected in the mature fish. Similar to the mammalian AMPA receptor, variants created by the alternate choice of flip and flop modules were found among transcripts of fGluR2 alpha-c and fGluR2 beta. The amino acid sequences of flip and flop modules of fGluR2 beta are identical to that of the rat GluR2, whereas the amino acid sequences of the flip and flop modules of fGluR2 alpha-c differ from the invariant consensus sequences of the rat AMPA receptor subunits. PMID- 8717348 TI - Sex differences in androgen receptor mRNA levels and regulation in hamster facial motoneurons. AB - We have previously shown that testosterone propionate augments hamster facial nerve regeneration to a greater extent in males than females. Further, sex differences in facial nerve regeneration have been observed. From those studies, we hypothesized that sex differences in nerve regeneration could be due to inherent differences in androgen receptor (AR) mRNA content within facial motor neurons (FMN) of male and female hamsters. In the present study, that hypothesis was tested using in situ hybridization, and computerized image analysis to quantify levels and regulation of AR mRNA in individual FMN of hamsters of both sexes. Intact and gonadectomized (gdx) male and female hamsters were used in the initial experiments in the study. In subsequent experiments, exogenous testosterone propionate (TP) was administered to the aforementioned groups of animals by subcutaneous implantation of one 10-mm Silastic capsule for 1, 2 or 7 days. FMN of intact females contained approximately 50% less AR mRNA than their male counterparts. Gonadectomy in males downregulated AR mRNA levels by approximately 50%, whereas no effects of gonadectomy were observed in females. Thus, in all paradigms where TP levels were low relative to the intact males, AR mRNA levels were approximately half of those in the intact male FMN. TP administration induced AR mRNA levels in gdx males within 1 day. Significant effects of TP were not detected in gdx females, and only after 7 days in the intact females. To our knowledge, the results of this study are the first quantitative demonstration of sex differences in steroid receptor mRNA content in a given neuronal population and substantiate the idea that sex differences in the effects of androgens on peripheral nerve regeneration are based on intrinsic sex differences in the levels and regulation of receptor mRNA in motor neurons. PMID- 8717349 TI - Localization of urokinase-type plasminogen activator mRNA in the adult mouse brain. AB - Urokinase-type plasminogen activator (uPA) is an inducible serine protease, secreted by a variety of cell types, that functions in fibrinolysis and has been implicated also in events such as cell migration and tissue remodeling and repair. To explore the role of uPA in the adult brain we have now screened the whole mouse brain for cells expressing the uPA gene through in situ hybridization using 35S-complementary RNA. uPA mRNA was visualized predominantly in three regions: (1) the subicular complex, (2) the entorhinal cortex, (3) the parietal cortex, where the signal was somewhat lower and confined to layers IV and VI. Weaker signals were seen in the basolateral nucleus of the amygdala and in the anterodorsal thalamic nucleus, and also in the hilus of the dentate gyrus where labeling was slightly over background. Cells exhibiting uPA mRNA signaling were large neurons according to morphological criteria. These results support the view of uPA being involved in neuronal functions of the adult brain, specifically in the hippocampal formation and the parietal cortex. PMID- 8717350 TI - MARCKS and protein F1/GAP-43 mRNA in chick brain: effects of imprinting. AB - The phosphorylation of MARCKS, but not protein F1/GAP-43, is increased in the intermediate and medial portion of the hyperstriatum ventrale (IMHV) after chick imprinting. Here we investigated if MARCKS, but not F1/GAP-43, gene expression would also be altered after imprinting. We first investigated the constitutive mRNA distribution of MARCKS and F1/GAP-43 in chick brain. MARCKS mRNA was expressed in most cells and exhibited a relatively homogeneous distribution. In contrast, F1/GAP-43 mRNA levels were elevated in discrete brain regions, as we had observed in mammals. The highest F1/GAP-43 mRNA levels in the chick brain were in sensory and associational structures such as the hyperstriatal complex and neostriatum, and lower levels were in structures involved in motor control, such as paleostriatum. These results in chick are consistent with the previously drawn generalization that F1/GAP-43 mRNA is expressed in those brain regions which exhibit synaptic plasticity. After imprinting, MARCKS mRNA levels in IMHV were higher in good learners than poor learners. In contrast, analysis of F1/GAP 43 mRNA levels revealed no differences related to training in any brain region sampled. These selective results for MARCKS but not F1/GAP-43 parallel the prior findings on their phosphorylation, and are consistent with our hypothesis that the very same proteins that are post-translationally modified in association with learning and memory also undergo alterations in their gene expression. PMID- 8717351 TI - Expression of the proenkephalin A gene in organotypic cultures of neocortex from newborn rats. AB - In rats, the proenkephalin A gene is expressed in proliferating cells of the neuroepithelial zone which later give rise to neocortical neurones and glial cells. Therefore, organotypic cultures of neocortex of newborn rats were used in the present study to examine whether neurones as well as glial cells expressed the gene. The slices were prepared at birth and kept in culture for 7-13 days. Proenkephalin mRNA was visualised by in situ hybridisation, while immunocytochemical staining for MAP-2 and GFAP was used to identify neurones and astroglial cells, respectively. In the analysed slices, only neurones contained proenkephalin mRNA. Activation of protein kinase C with tetradecanoylphorbol acetate (1 mumol/l) caused a strong increase in the number of neurones expressing proenkephalin mRNA. Our results indicate that a large number of neurones is able to express the proenkephalin gene under these conditions. However, only a few of them have a basal expression which is strong enough to be detected with in situ hybridisation. PMID- 8717353 TI - Distinct structure-activity relations for stimulation of 45Ca uptake and for high affinity binding in cultured rat dorsal root ganglion neurons and dorsal root ganglion membranes. AB - The [3H]resiniferatoxin (RTX) binding assay using membrane preparations has been used to identify and characterize the vanilloid receptors in the central and peripheral nervous system of different species. In the present study, using cultured adult rat dorsal root ganglion neurons either in suspension or attached to the tissue culture plates, we developed an assay to measure specific [3H]RTX binding by the intact cells. We were able to characterize the vanilloid binding characteristics of the neurons and compared those to the properties of vanilloid binding sites present in rat dorsal root ganglia membrane preparations. We found that [3H]RTX bound with similar affinity and positive cooperativity to attached neurons (cultured for 5 days before being assayed), neurons in suspension (using a filtration assay) and dorsal root ganglion membrane preparations. Dissociation constants obtained in the three assays were 47.6 +/- 3.5 pM, 38.4 +/- 3.1 pM and 42.6 +/- 3.1 pM, respectively. The cooperativity indexes determined by fitting the data to the Hill equation were 1.73 +/- 0.11, 1.78 +/- 0.12 and 1.78 +/- 0.09, respectively. The maximal binding capacity was 0.218 +/- 0.026 fmol/10(3) cells and 0.196 +/- 0.021 fmol/10(3) cells in the case of the attached cells and cells in suspension, respectively. Nonradioactive RTX, capsaicin, capsazepine and resiniferonol 20-homovanillylamide fully displaced specifically bound [3H]RTX from cells in suspension with Ki and Hill coefficient values of 42.5 +/- 5.3 pM, 2.06 +/- 0.16 microM, 3.16 +/- 0.21 microM and 32.4 +/- 4.1 nM and 1.79 +/- 0.17, 1.68 +/- 0.06, 1.72 +/- 0.11 and 1.81 +/- 0.12, respectively. Structure-activity analysis of different vanilloid derivatives revealed that the various compounds have distinct potencies for receptor binding and inducing 45Ca uptake in rat dorsal root ganglion neurons. Affinities for receptor binding and stimulation of 45Ca uptake of RTX, resiniferonol 20-homovanillylamide, RTX-thiourea, tinyatoxin, phorbol 12,13-dibenzoate 20-homovanillylamide and capsaicin were 38.5 +/- 2.9 pM, 25.7 +/- 3.0 nM, 68.5 +/- 3.8 nM, 173 +/- 25 pM, 7.98 +/- 0.83 microM and 4.93 +/ 0.35 microM as compared to 0.94 +/- 0.12 nM, 26.5 +/- 3.5 nM, 149 +/- 30 nM, 1.46 +/- 0.25 nM, 1.41 +/- 0.48 microM and 340 +/- 57 nM. Computer fitting of the data yielded Hill coefficient values indicating positive cooperativity of receptor binding; however, stimulation of 45Ca uptake appeared to follow a non cooperative mechanism of action. The competitive capsaicin antagonist capsazepine inhibited specific binding of [3H]RTX by rat dorsal root ganglion membrane preparations with Ki and Hill coefficient values of 3.89 +/- 0.38 microM and 1.74 +/- 0.11. On the other hand it inhibited the induction of 45Ca uptake into the cells induced by capsaicin and RTX in a non-cooperative fashion with Ki values of 271 +/- 29 nM and 325 +/- 47 nM. Our results show that the membrane binding assay relates to the reality of receptor function in the intact, cultured neurons, both in terms of affinity and positive cooperativity. However the different vanilloid derivatives displayed markedly distinct structure-activity relations for high affinity receptor binding and stimulation of 45Ca uptake into rat dorsal root ganglion neurons. Among various explanations for this discrepancy, we favor the possibility that the two assays detect distinct classes of the vanilloid (capsaicin) receptor present in primary sensory neurons. PMID- 8717352 TI - Promoter elements and second messenger pathways involved in transcriptional activation of tyrosine hydroxylase by ionomycin. AB - Membrane depolarization, or agents which increase intracellular calcium, elicit transcriptional activation of tyrosine hydroxylase (TH). In this study we analyze the factors involved in the regulation of the TH promoter by a calcium ionophore. PC12 cells were transiently transfected with plasmids containing wild type or mutated 5' flanking sequences of the rat TH gene, fused to bacterial chloramphenicol acetyl transferase (CAT). Point mutations introduced into the consensus cAMP-regulatory element (CRE) abolished the induction of CAT by ionomycin indicating that it is essential for mediating the calcium response. An intact and functional AP1 site did not confer calcium inducibility when the CRE/CaRE sequence was mutated. The extent and kinetics of the increase in intracellular calcium as well as the induction of CAT activity under the control of TH promoter by ionomycin were similar in PC12 cells and in the A123.7, protein kinase A (PKA) deficient cell line. In both cell lines addition of ionomycin rapidly increased the phosphorylation of transcription factor CREB at Ser-133. These results suggest that the activation of TH transcription by ionomycin does not require PKA. However, KN62 an inhibitor of Ca2+/calmodulin dependent (CaM) kinases prevented the induction indicating possible involvement of CAM kinases in the calcium response. PMID- 8717354 TI - Caffeine-induced expression of c-fos mRNA and NGFI-A mRNA in caudate putamen and in nucleus accumbens are differentially affected by the N-methyl-D-aspartate receptor antagonist MK-801. AB - Caffeine (100 mg/kg, i.p.) induces a rapid increase in the expression of mRNA for the immediate early genes (IEGs) c-fos and NGFI-A in rat striatum. We have examined how this response is affected by pretreatment with either the noncompetitive N-methyl-D-aspartate (NMDA) receptor antagonist MK-801 (1 and 3 mg/kg, i.p.), the competitive NMDA receptor antagonist D-CPP (6 mg/kg, i.p.), or the non-selective excitatory amino acid receptor antagonist kynurenic acid (300 mg/kg, i.p). The two NMDA receptor antagonists significantly reduced the caffeine induced expression of both c-fos mRNA and NGFI-A mRNA in the medial part of the caudate putamen. The effect was less pronounced in the lateral part of the caudate putamen. MK-801 caused an enhancement of c-fos and NGFI-A mRNA expression in nucleus accumbens. Pretreatment with kynurenic acid caused no marked alterations in the caffeine-induced expression of c-fos mRNA and NGFI-A mRNA in any brain region. These findings suggest that glutamatergic transmission via NMDA receptors contributes to the induction of c-fos mRNA and NGFI-A mRNA by caffeine in striatum. In addition we show that MK-801 can either increase or decrease the caffeine effect on IEGs depending on the region studied. PMID- 8717355 TI - Circadian variation of EAAC1 glutamate transporter messenger RNA in the rat suprachiasmatic nuclei. AB - Using in situ hybridization, we examined temporal changes of the EAAC1 glutamate transporter mRNA within the suprachiasmatic nuclei (SCN) of rats in constant darkness. Film autoradiographs showed that the SCN and supraoptic nuclei (SON) contained a marked density of hybridization signal. Analysis of silver grains per cell in emulsion-dipped sections indicated that cellular expression of EAAC1 mRNA in the SCN was elevated during the latter part of the subjective night and at the beginning of the subjective day, with a peak at circadian time 23.1 as determined by cosinor analysis. The times at which EAAC1 mRNA is highest correspond to the time points at which extracellular glutamate, a neurotransmitter that putatively mediates photic entrainment, has been reported to be low within the SCN. The presence of EAAC1 mRNA in the SCN and SON may partially explain the resistance of these nuclei to glutamate receptor-mediated excitotoxins; furthermore, the raised level preceding subjective dawn in the SCN may ensure sub-toxic levels of extracellular glutamate at the onset of photic stimulation during the LD cycle. In contrast, cellular expression of EAAC1 mRNA in the cingulate cortex and reticular thalamus remained constant at all time points studied. These results suggest that there is circadian control of the EAAC1 mRNA by the clock intrinsic to the SCN. PMID- 8717356 TI - Dopaminergic regulation of striatonigral tachykinin and dynorphin gene expression: a study with the dopamine uptake inhibitor GBR-12909. AB - The present study examined the modulatory role of dopamine (DA) on striatonigral preprotachykinin (PPT) and prodynorphin (PD) gene expression, employing the DA uptake inhibitor, GBR-12909 (GBR), as a tool. The striatal and nigral levels of tachykinin (substance P (SP), neurokinin A (NKA)) and dynorphin (dynorphin A(1-8) (DYN)) peptides were determined by radioimmunoassays. The abundance of mRNAs in the striatum was quantified by Northern blot analysis. The rate of transcription of PPT and PD genes in the striatum was measured by transcription run-on assays. A regimen of repeated administration of GBR (20 mg/kg/day, i.p., for 1-4 days) to female Sprague-Dawley rats increased striatal and nigral SP, NKA, and DYN peptide levels. The increased peptide levels were associated with increases in the abundance of PD mRNA and PPT mRNA and increases in the rate of transcription of PD and PPT genes in the striatum, suggesting a GBR-induced activation of the striatonigral tachykinin and dynorphin neurons. Dopaminergic denervation with 6 hydroxydopamine (6OHDA) blocked the GBR-induced increases in SP and DYN and PPT and PD mRNAs. The concurrent administration of the D1 DA antagonist, SCH-23390, blocked the GBR-induced increases in SP, NKA and PPT mRNA but failed to affect DYN or PD mRNA levels; the concurrent administration of the D2 DA antagonist, spiperone, blocked the GBR-induced increases in SP, NKA and PPT mRNA and also DYN and PD mRNA. The study reveals that repeated administration of GBR enhances the levels of tachykinin and dynorphin peptides in striatonigral neurons by a stimulus-transcription-biosynthesis coupling mechanism. The GBR-induced effects are dependent on the integrity of nigrostriatal dopaminergic neurons and the presence of D1 and/or D2 DA receptors. PMID- 8717357 TI - The human glycine receptor beta subunit: primary structure, functional characterisation and chromosomal localisation of the human and murine genes. AB - The inhibitory glycine receptor (GlyR) is a pentameric receptor comprised of alpha and beta subunits, of which the beta subunit has not been characterised in humans. A 2106 bp cDNA, isolated from a human hippocampal cDNA library, contained an open reading frame of 497 amino acids which encodes the beta subunit of the human GlyR. The mature human GlyR beta polypeptide displays 99% amino acid identity with the rat GlyR beta subunit and 48% identity with the human GlyR alpha 1 subunit. Neither [3H]strychnine binding nor glycine-gated currents were detected when the human GlyR beta subunit cDNA was expressed in the human embryonic kidney 293 cell line. However, co-expression of the beta subunit cDNA with the alpha 1 subunit cDNA resulted in expression of functional GlyRs which showed a 4-fold reduction in the EC50 values when compared to alpha 1 homomeric GlyRs. Glycine-gated currents of alpha 1/beta GlyRs were 17-fold less sensitive than homomeric alpha 1 GlyRs to the antagonists picrotoxin, picrotoxinin and picrotin, providing clear evidence that heteromeric alpha 1/beta GlyRs were expressed. The beta subunit appears to play a structural rather than ligand binding role in GlyR function. Fluorescence in situ hybridisation was used to localise the gene encoding the human GlyR beta subunit (GLRB) to chromosome 4q32, a position syntenic with mouse chromosome 3. In situ hybridisation using the human GlyR beta subunit cDNA showed that the murine GlyR beta subunit gene (Glrb) maps to the spastic (spa) locus on mouse chromosome 3 at bands E3-F1. This is consistent with the recent finding that a mutation in the murine GlyR beta subunit causes the spa phenotype. It also raises the possibility that mutations in the human beta subunit gene may cause inherited disorders of the startle response. PMID- 8717358 TI - Evidence for the involvement of non-androgenic testicular factors in the regulation of hypothalamic somatostatin and GHRH mRNA levels. AB - The growth hormone (GH) secretory pattern is dependent on sex and developmental stage. It is generally accepted that in the male rat this pattern is markedly influenced by androgens secreted by the Leydig cells. Recent findings, however, point to the existence of other non-androgenic testicular factors produced by the Sertoli cells and which regulate in vivo the GH responses to growth hormone releasing hormone (GHRH). The aim of this work was to investigate the role played by non-androgenic testicular factors on hypothalamic somatostatin (SST) and GHRH mRNA levels. Seventy-day-old male Sprague-Dawley rats were used throughout the work. They were divided into five groups: (1) control rats; (2) gonadectomized rats; (3) gonadectomized rats supplemented with exogenous administration of dihydrotestosterone (DHT); (4) ethylene dimethane sulphonate (EDS)-treated rats; (5) EDS-treated rats supplemented with exogenous administration of DHT. EDS is a cytotoxic agent that specifically destroys the Leydig cells. The rats were killed after 15 days of treatment. Hypothalamic SST mRNA levels were determined by Northern blot and by in situ hybridization, and GHRH mRNA levels assessed by Northern blot. We found that selective removal of Leydig cells with EDS greatly reduced the SST mRNA content in the periventricular nucleus of the hypothalamus. These levels were significantly lower than those found in gonadectomized rats. Furthermore, replacement treatment with dihydrotesterone (DHT) did not completely restore SST mRNA levels in EDS-treated rats, contrasting with the complete recovery of SST mRNA levels in gonadectomized rats. On the other hand, gonadectomy and EDS treatment produced a significant reduction in GHRH mRNA levels. DHT administration reversed the action of gonadectomy, but did not restore GHRH mRNA content in EDS-treated rats. These data suggest that, in addition to testosterone, as yet unidentified non-androgenic testicular factors can significantly influence SST and GHRH mRNA levels. This may indicate that non androgenic testicular factors acting at hypothalamic level may be important in the neuroregulation of GH secretion and in the maintenance of sexual dimorphism in GH secretory pattern. PMID- 8717359 TI - A herpes simplex virus-1 vector containing the rat tyrosine hydroxylase promoter directs cell type-specific expression of beta-galactosidase in cultured rat peripheral neurons. AB - A defective herpes simplex virus-1 (HSV-1) vector system was used to study cell type-specific expression of the tyrosine hydroxylase (TH) gene. HSV-1 particles containing 663 bp (pTHlac 663), 278 bp (pTHlac 278), or 181 bp (pTHlac 181) of the rat TH promoter driving E. coli LacZ were used to infect superior cervical ganglia (SCG: TH-expressing tissue) and dorsal root ganglia (DRG:non-TH expressing tissue) cultures. One day after infection, expression of beta galactosidase was visualized by X-gal cytochemistry. Following viral transduction with pTHlac 663 at a multiplicity of infection of 0.2, 14.4% of the SCG neurons were X-gal positive whereas only about 0.9% of DRG neurons were X-gal positive. Infection with either pTHlac278 or 181 resulted in 3-fold more X-gal-positive DRG neurons. These results suggest that (i) the defective HSV-1 vector system may be useful in defining regulatory promoter motifs; (ii) 663 bp of the rat TH promoter contains sufficient information for cell type-specific expression in peripheral nervous system neurons; and (iii) sequences between -278 and -663 contain an element(s) that represses gene expression in non-catecholamingeric neurons. PMID- 8717360 TI - The effects of D1 or D2 dopamine receptor blockade on zif/268 and preprodynorphin gene expression in rat forebrain following a short-term cocaine binge. AB - Selective D1 or D2 dopamine receptor antagonists were used to investigate the transynaptic regulation of mRNAs coding for the opioid peptide, preprodynorphin, and the nuclear transcription factor, zif/268 after an acute cocaine binge. Rats were injected intraperitoneally with the D1 receptor antagonist, SCH 23390, or the D2 receptor antagonist, sulpiride, 30 min prior to 3 hourly injections of saline or 20 mg/kg cocaine and killed 1 h after the final injection. Behavioral ratings indicated that SCH 23390 blocked, whereas sulpiride augmented, cocaine induced stereotypical behaviors. Striatal sections were hybridized with oligonucleotides coding for zif/268 and preprodynorphin. Quantitative image analysis of autoradiograms revealed that (1) SCH 23390 completely suppressed basal and cocaine binge-induced zif/268 mRNA in the striatal and cerebral cortical areas examined; (2) sulpiride enhanced basal levels of zif/268 mRNA in the medial caudate and dorsomedial shell of the nucleus accumbens; (3) sulpiride partially blocked cocaine binge-induced levels of zif/268 mRNA in the dorsal striatum but had no effect in sensory cortex; (4) SCH 23390, but not sulpiride, significantly reduced the constitutive expression of preprodynorphin mRNA; and (5) SCH 23390 and sulpiride blocked cocaine binge-induced expression of preprodynorphin mRNA in the dorsal striatum. PMID- 8717361 TI - Neuropeptide Y gene expression and receptor autoradiography in hypertensive and normotensive rat brain. AB - Neurones containing neuropeptide Y (NPY) may participate in central cardiovascular control by tonically influencing barosensitive neurones within the nucleus tractus solitarius. The present study has employed both in situ hybridisation histochemistry and receptor autoradiography, to visualise the expression of prepro-NPY mRNA in the forebrain and to determine the NPY receptor subtype(s) in the brainstem, respectively. Prepro-NPY gene expression was visualised in the hypothalamus, cortex, dentate gyrus and lateral reticular thalamus from age-matched spontaneously hypertensive rats (SHR) and normotensive Don Ryu rats (DRY) and Wistar Kyoto rats (WKY). Quantitative densitometry revealed an increase in the NPY transcript in the arcuate nucleus of SHR rats compared to their normotensive counterparts. Autoradiography using [125I]Bolton Hunter-NPY (BH-NPY, 15 pM) demonstrated NPY binding sites in the area postrema, the commissural nucleus tractus solitarius (cNTS) and the inferior olivary complex. NPY (1 microM) and peptide YY (1 microM), but not [Leu31,Pro34]NPY (10 100 nM), fully inhibited the binding of [125I]BH-NPY. These results indicate that NPY receptors of the Y2 subtype predominate in the dorsal vagal complex. Unilateral nodose ganglionectomy resulted in a partial loss of NPY binding sites in the commissural NTS, but not the area postrema, suggesting that a proportion of binding sites (Y2 subtype) are present on central vagal terminals. While all three rat strains appear to have the same relative proportions of NPY receptor subtypes in the brainstem, the relevance of the differential NPY gene expression in the arcuate nucleus regarding central cardiovascular control mechanisms and/or the pathogenesis of hypertension remains to be elucidated. PMID- 8717362 TI - Changes in expression of lymphocyte amyloid precursor protein mRNA isoforms in normal aging and Alzheimer's disease. AB - We measured, by employing a quantitative reverse-transcriptase PCR procedure, the relative (to beta-actin) levels of amyloid precursor protein APP751 and APP770 mRNA isoforms in lymphocytes obtained from 64 cognitively intact subjects ranging in ages from 20 to 91 years and in 19 patients with sporadic Alzheimer's disease. A positive correlation was observed between the relative lymphocyte APP751 mRNA levels and subject age for the cognitively intact cohort. No difference in lymphocyte APP751 mRNA levels was observed between Alzheimer's disease patients and their age-matched controls (> 55 years of age). However, the ratio of lymphocyte APP751:APP770 mRNA levels was significantly lower in Alzheimer's disease subjects compared to the > 55-year-old cohort. This decreased ratio is most likely due to an average 31% increase in the lymphocyte APP770 isoform in Alzheimer's disease patients compared to 12% in the > 55-year-old cognitively intact group. Marked individual differences in amount of APP mRNA isoforms were encountered among all the subject groups and in the < or = 55-year-old cohort, a 10-fold variation in individual APP751 mRNA levels was observed. The relevance of these findings in lymphocytes to the pathogenesis of Alzheimer's disease is discussed. PMID- 8717363 TI - Differential expression of alpha-CGRP and beta-CGRP genes within hypoglossal motoneurons in response to axotomy. AB - In this study we have analysed, by in situ hybridization, the expression of the genes for both alpha-CGRP and beta-CGRP in hypoglossal motor nuclei following transection of the left hypoglossal nerve. Our results show that the gene for alpha-CGRP displays a peculiar sequence of regulation (a successive up-down-up recovery sequence) within ipsilateral hypoglossal motoneurons in response to axotomy. It is initially up-regulated, then down-regulated (displaying mRNA levels below basal), and later again up-regulated before recovery. By contrast, the gene for beta-CGRP displays a successive and distinct up-down-recovery sequence of regulation (it does not display a second increase in mRNA production). The first up-regulation of the alpha-CGRP gene occurs just during the early period of perineuronal glial reaction and the second up-regulation just during the period of delayed astrocyte reaction and muscle reinnervation. Because alpha-CGRP is a neuron-derived factor for many types of cells, including astrocytes and skeletal myocytes, our results suggest that the pleiotropic alpha CGRP may be a motoneuron-derived trophic signal for both glial and skeletal muscle cells in order to maintain the motoneuron itself and, in consequence, might be of therapeutic interest in treating degenerative disease of motoneurons. beta-CGRP might be redundant within the hypoglossal motoneurons. PMID- 8717364 TI - Changes in cholecystokinin mRNA expression after amygdala kindled seizures: an in situ hybridization study. AB - Cholecystokinin (CCK) can be a potent anticonvulsant neuropeptide in certain seizure models. Therefore, we examined whether seizures produced by electrical kindling of the amygdala or electroconvulsive seizures (ECS) would affect the expression of CCK mRNA in rat brain. Following a single kindled seizure, CCK mRNA expression was decreased about 20-58% in the amygdala. In contrast, after multiple consecutive kindled seizures, CCK mRNA expression was increased in the amygdala, cerebral cortex, CA1 pyramidal cell layer of the hippocampus and dentate hilus. A single ECS produced no effect on CCK mRNA expression, but multiple ECS increased expression in the interneurons of the hippocampus 24 h after the last seizure. Since seizures produced by ECS can be anticonvulsant to further ECS or kindled seizures, the CCK increases in the hippocampus may represent a compensatory anticonvulsant adaptation observed in both models. Overall, the kindling-induced alterations in CCK expression appear to be more complex involving multiple brain regions and distinct temporal properties. PMID- 8717365 TI - The RNA binding protein HuD: rat cDNA and analysis of the alternative spliced mRNA in neuronal differentiating cell lines P19 and PC12. AB - HuD belongs to a family of neurospecific RNA binding proteins found in man, frog and fly [49]. To investigate whether this protein is involved in regulation of neuronal differentiation of rodent cells in vivo and in vitro, the cDNA of the rat homolog gene (r-HuD) was cloned, its expression was studied in rat brain and in neurogenic cell lines, and the splicing of its RNA was analyzed. Coding sequences of HuD from man and rat were found to be 99.5 and 95% identical at protein and DNA level, respectively. In rat brain r-HuD transcripts 3.7 and 4.2 kb in length were detected by Northern blot analysis. RT-PCR and in situ hybridization revealed that rodent homologues of HuD transcripts are present in P19 mouse embryo carcinoma and in PC12 rat pheochromocytoma cell lines both able to differentiate into neurons. In contrast, r-HuD transcripts were not detectable in the rat glioma cell line C6. In P19 cells a strong induction of HuD mRNA was observed after triggering neuronal differentiation by retinoic acid, whereas in PC12 cells the mRNA was present before and after nerve growth factor (NGF) induced neuronal differentiation. In both neuronal cell lines and in brain of adult rat and mouse HuD mRNA is alternatively spliced in a region which encodes a proline rich linker domain between the second and third RNA recognition motif. This RNA processing event seems to be differently regulated in PC12 cells on the one hand, and in P19 cells and brain of rat and mouse on the other. PMID- 8717366 TI - Anesthesia during hormone administration abolishes the estrogen induction of preproenkephalin mRNA in ventromedial hypothalamus of female rats. AB - Estrogen treatment increases preproenkephalin (PPE) mRNA levels in the ventromedial nucleus of the hypothalamus (VMH). Roy et al. (Brain Res., 337 (1985) 163-166) discovered that anesthesia during estrogen priming could reduce female rat sexual receptivity. In the present study we tested whether the action of estrogen to induce PPE gene expression in the VMH could be similarly affected by anesthesia. By quantitative in situ hybridization and slot-blot analysis techniques we found a 1.8-fold increase in PPE mRNA levels in the VMH after 1 hour of estrogen treatment in ovariectomized (OVX) Sprague-Dawley female rats. Anesthetizing the rats with pentobarbital for 1 h during the exposure to estrogen blocked the estrogen induction of PPE mRNA in the VMH. By way of contrast no changes in the PPE mRNA levels were observed in the caudate putamen. A similar trend was seen using chloral hydrate. It appears that neuronal activity is required for the early phase of estrogen induction of PPE mRNA levels in the VMH. This in turn could be correlated with changes in female sociosexual behaviors. PMID- 8717367 TI - The helix-loop-helix transcription factor USF interacts with the basal promoter of human amyloid precursor protein. AB - Nuclear factors from HeLa, PC12, NG108-15 and SK-N-SH cell lines recognized an oligonucleotide (-56 to -37: APP-E1) containing an E box (CANNTG) which had previously been characterized as a DNase I-protected sequence in the basal promoter of the human amyloid precursor protein (APP) gene. Binding to APP-E1 was competed with an oligonucleotide encompassing the recognition site of the transcription factor USF. Antibodies directed against USF interacted with the APP E1-protein complex and in vitro synthesized USF could bind APP-E1. Co-expression of USF cDNA transactivated a human APP-reporter gene construct. These results suggest that USF may play a role in the expression of the APP gene. PMID- 8717368 TI - Antisense oligonucleotide to NOR-1, a novel orphan nuclear receptor, induces migration and neurite extension of cultured forebrain cells. AB - We previously identified a novel orphan nuclear receptor referred to as NOR-1 from rat forebrain cells. This study examined the role of NOR-1 in primary cultured forebrain cells by selectively inhibiting NOR-1 expression by addition of antisense oligonucleotide to the culture media. Treating cells with the antisense oligomer resulted in the following dramatic morphological changes: (i) cell migration, (ii) extension of processes, and (iii) formation of cellular aggregates. Immunocytochemistry for microtubule-associated protein 2 revealed that the processes were filled with neurites growing from neuronal cells. These findings suggest that NOR-1 may be involved in the molecular mechanisms regulating neural differentiation. PMID- 8717369 TI - Induction of SPI-3 mRNA, encoding a serine protease inhibitor, in gerbil hippocampus after transient forebrain ischemia. AB - We cloned genes the expression of which is induced in the Mongolian gerbil (Meriones unguiculatus) hippocampus after transient forebrain ischemia by a differential display technique. Among these genes, a rat serine protease inhibitor SPI-3 homologue was isolated. Present analyses suggested that the expression of gerbil SPI-3 mRNA was closely associated with delayed neuronal death and may block activities of proteases leaking from degenerating neurons or may support neuronal survival. PMID- 8717370 TI - Expression of Na+/myo-inositol cotransporter mRNA in the inner ear of the rat. AB - We have demonstrated the cellular localization of Na+/myo-inositol cotransporter (SMIT) mRNA in the rat inner ear by in situ hybridization. In the cochlea, the most intense SMIT mRNA signals were observed in fibrocytes of the spiral ligament, moderate signals were found in the spiral limbus, inner hair cells and spiral ganglion cells, while the hybridization signals were almost undetectable in the marginal cells of the stria vascularis and outer hair cells. In the vestibular system, moderate hybridization signals were found in the sensory epithelium, fibrocytes and vestibular ganglion cells. These findings suggest that SMIT plays an important role in maintenance of intracellular ionic balance and cell volume in the inner ear, especially in the fibrocytes associated with generation of the ion gradients between the endolymph and perilymph. PMID- 8717371 TI - Heat shock factor-1 mediates the transcriptional activation of Alzheimer's beta amyloid precursor protein gene in response to stress. AB - Stress may be involved in the pathogenesis of Alzheimer's disease. There is a heat shock element located at position -317 bp on the beta-amyloid precursor protein (beta-APP) gene promoter. Recently we demonstrated [4] that stress, in the form of heat shock, ethanol and sodium arsenite treatment, transcriptionally activates the beta-APP gene. In this report we demonstrate that the nuclear factor that mediates this activation is heat shock factor-1. PMID- 8717372 TI - Expression of tachykinin NK1 receptor mRNA in dorsal root ganglia of the mouse. AB - We examined whether mRNA coding for tachykinin NK1 receptor is expressed in the dorsal root ganglion (DRG) of the mouse, using reverse transcription-polymerase chain reaction (RT-PCR). Both the RT-PCR of the total RNA from the DRGs using four pairs of primers and the digestion of these products with the restriction enzymes gave bands with the predicted length. Further amplification (nested PCR) of part of one PCR product also gave a band with the predicted length. Southern blot hybridization of RT-PCR products of total RNA from the DRG and several CNS regions revealed that the expression level of NK1 receptor mRNA in the DRG was similar to the cerebellum and less than the olfactory bulb, cerebral cortex, medulla oblongata and spinal cord. The present results suggest that NK1 receptor mRNA is expressed in the mouse DRG, although the level is relatively low. PMID- 8717373 TI - Expression of Na+/myo-inositol cotransporter mRNA in normal and hypertonic stress rat eyes. AB - We studied the localization of Na+/myo-inositol cotransporter (SMIT) mRNA in normal and hypertonic stress rat eyes by in situ hybridization histochemistry using cRNA probes. SMIT mRNA signals were observed in the iris-ciliary body, the lens epithelial cells, and the ganglion cell layer and the inner nuclear layer of the retina. There was a rapid increase on SMIT mRNA in the retina of hypertonic stress rats compared with control rats. These findings suggest that Na+/myo inositol cotransporter gene expression is osmotically regulated in vivo to protect retinal neuronal function against hypertonic stress. PMID- 8717375 TI - Improved lipid-mediated gene transfer into primary cultures of hippocampal neurons. AB - We have examined lipids as transfection agents to introduce recombinant plasmids into primary cultures of rat hippocampal neurons. By modifying the protocol for transfection mediated by the commercial reagent DOTAP, we were able to achieve a transfection efficiency of about 3%. Expression of various transfected gene products was sustained for several weeks in culture, the neurons developed normally and the transfected gene products were targeted to the appropriate subcellular compartment. PMID- 8717374 TI - Induction of PGH synthase and c-fos mRNA during early reperfusion of ischemic rat brain. AB - We examined the effect of reversible ischemia on the transcription of prostaglandin endoperoxide synthase (PGHS-1) and c-fos mRNA in rat cerebral cortex. The level of PGHS-1 mRNA climaxed after 30 min of ischemia whereas transcription of c-fos mRNA peaked after 60 min of postischemic reperfusion. We conclude that cerebral ischemia causes early transcription of PGHS-1, without modulation by the c-fos gene or its translated product. PMID- 8717376 TI - Repeated ECS induces GluR1 mRNA but not NMDAR1A-G mRNA in the rat hippocampus. AB - The neural basis underlying the cognitive side effects of ECT is unknown. Recent studies suggest that the memory dysfunction may be caused by alterations in hippocampal synaptic efficacy [20]. In situ hybridisation was used to examine the possible receptor mechanisms responsible for this effect. Repeated ECS markedly increased mRNA expression for the GluR1 subunit of the AMPA receptor, but not the NMDAR1A-G subtypes of the NMDA receptor, relative to control treatments. This effect was present 24 h after the last seizure and may be responsible for the expression of the ECS-induced increase in synaptic efficacy. PMID- 8717377 TI - Identification and partial characterization of the high-affinity choline carrier from rat brain striatum. AB - [3H]Choline mustard aziridinium ion binds irreversibly to the sodium-coupled high affinity choline transport protein in a sodium-dependent and hemicholinium sensitive manner, and thus is a useful affinity ligand. In rat striatal synaptosomal membranes, it radiolabels two polypeptides with apparent molecular masses of 58 and 35 kDa. Based upon the use of two different experimental approaches, it appears that neither of these polypeptides is glycosylated. PMID- 8717378 TI - New approaches to vaccine development. PMID- 8717379 TI - Towards a strategy of universally efficacious vaccination against pathogens uniquely susceptible to cell-mediated attack. AB - Infection by some intracellular parasites is contained only by cell-mediated immunity, and yet antibody is produced at the expense of the cell-mediated response upon natural infection, leading to chronic or fatal disease. Effective vaccination must therefore generate an immunological imprint ensuring a strong and stable cell-mediated response upon infection. Such diseases include leprosy, tuberculosis, the leishmaniasis and AIDS (Kaplan and Cohn (1986) Int. Rev. Exp. Pathol. 28, 45-78; Surcel et al. (1994) Immunology 81, 171-176; Pearson et al. (1983) Rev. Infect. Dis. 5, 907-927; Clerici and Shearer (1993) Immunol. Today 14, 107-111). BALB/C mice are susceptible to Leishmania major, a protozoan that causes cutaneous leishmaniasis in man, by the criterion that substantial infection results in antibody production and progressive disease (Locksley and Scott (1991) Immunoparasitology Today, A58-A61; J.N. Menon and P.A. Bretscher, unpublished data). Infection of BALB/C mice with very few parasites results in an exclusive cell-mediated, Th1-like response and resistance to an ordinarily pathogenic, high dose challenge. This resistance is associated with a strong and stable cell-mediated response (Bretscher et al. (1992) Science 257, 539-542; J.N. Menon and P.A. Bretscher, unpublished data). The generation of this Th1 imprint by low dose infection has been achieved with three very different strains of the parasite. There is a similar dependency of susceptibility and resistance on relative parasite dose in 'susceptible' and 'resistant' mice and in mice of 'intermediate susceptibility'. For example, 'resistant' mice are resistant to substantial infection but succumb to infection with very high doses of parasites. We therefore propose that infection of a genetically diverse population with a very low dose of viable parasites, that does not induce antibody in any individual, will either induce cell-mediated immunity and contain the parasite, or the parasite will grow until it reaches the threshold required to induce cell mediated immunity, thereby generating the required imprint. Low dose infection may thus constitute universally efficacious vaccination. The pertinence of these observations to improving the efficacy of BCG vaccination against tuberculosis is discussed. PMID- 8717380 TI - Antigen handling in the gastrointestinal lamina propria. AB - Enteric infections associated with bacteria, viruses and parasites are a world health problem accounting for at least one billion episodes of diarrhoea and up to 10 million deaths in the Third World each year. The widely held conclusion that responses which are protective in the gastrointestinal tract are best stimulated by mucosal presentation of antigen remains largely unchallenged. Despite this there are but a few effective mucosal vaccines. The mucosal immune system of the gastrointestinal tract is presented with a contrasting array of antigens, ranging from harmless dietary components to highly pathogenic organisms, and has evolved a battery of responses from which an appropriate response may be orchestrated. We have postulated that the microenvironment in which an antigen is processed and presented to the mucosal immune system may profoundly influence the nature of the response and the purpose of this review is discuss a possible role for the lamina propria in this process. PMID- 8717381 TI - Manipulating systemic and mucosal immune responses with skin-deliverable adjuvants. AB - Most medically important bacterial and viral pathogens gain entry into the body either via the skin or a mucosal surface. Vaccination provides a viable and cost effective strategy for the prevention of such diseases and it has always been a principal aim with vaccinologists, to be able to promote simultaneously, protective immune responses both systemically and at mucosal surfaces. The paradigm that mucosal immunity is best stimulated by exposure to antigen via a mucosal route simply because inductive sites such as Peyer's patches and bronchial associated lymphoid tissues are located in the mucosal epithelium, has promoted a plethora of immunizing strategies aimed at delivering both antigen and adjuvant to mucosal surfaces. We have developed a novel adjuvant system capable of intradermal delivery of antigens complexed in an ISCOSOME delivery vehicle. This adjuvant, referred to as a skin and mucosal adjuvant or SAMA4, was efficacious in eliciting both systemic and mucosal IgG and IgA antibodies in sheep, pigs and mice. SAMA4 does not induce granulomatous lesions at the site of vaccine delivery and can be used to deliver adjuvanted antigens by other routes including intranasal, oral and intravaginal. Using ovalbumin as a test antigen, intradermally delivered ovalbumin-SAMA4 complexes was found to be very effective in promoting a cytotoxic T cell response. Attempts to dissect the mode of action of SAMA4 by flow cytometric analysis of lymphocyte populations from the spleen, lung, liver and thymus revealed an effect of route of vaccine delivery upon the composition of specific lymphocyte subsets in these various organ compartments. From this, it can be inferred that SAMA4 induced a route-dependent re mobilization and alteration in lymphocyte trafficking patterns. Other mucosal adjuvants such as cholera toxin B and microspheres, when injected intradermally, tended to promote primarily, an IgG and not an IgA response against hte carrier antigen. PMID- 8717382 TI - Clinical and immunological characteristics of the emulsion form of inactivated influenza vaccine delivered by oral immunization. AB - Prophylaxis of human respiratory diseases caused by influenza viruses is actually a problem of infectious pathology because of their wide prevalence. In our investigations, safety, reactogenicity and immunological activity of the orally administered emulsion-inactivated influenza vaccine prepared from influenza virus strains of types A(H1N1), A(H3N2) and B have been studied. Clinical studies of the emulsion-inactivated influenza vaccine on volunteers has shown its safety and nonreactogenicity. The orally administered vaccine did not cause weak, middle or strong general or local reactions including clinical, biochemical, haemotological and immunological reactions. The emulsion-inactivated vaccine has high immunological activity and induces reliable increases in the level of secretory immunoglobulin A to influenza viruses A and B in protective titers in nasal secretions and saliva of volunteers after one oral administration. The obtained results indicate the expediency of further investigation and improvement of inactivated influenza vaccine for oral administration. PMID- 8717383 TI - Poly(lactide-co-glycolide) microencapsulation of vaccine antigens. AB - Fimbriae from Bordetella pertussis have been encapsulated in poly(lactide-co glycolide) (PLG) microspheres of a size appropriate for oral administration. The binding of antibodies which react with conformational or linear fimbrial epitopes, to fimbriae released from microspheres, suggested that the process of was not detrimental to the native integrity of the protein. Mice were immunised by oral gavage with a single dose of microencapsulated fimbriae, or with fimbriae adsorbed onto alhydrogel and administered by intraperitoneal injection. The resulting immune responses in serum were comparable but only oral administration of microencapsulated fimbriae elicited specific immune responses in external secretions. Six weeks after immunisation, both groups of immunised animals were protected against challenge with live B. pertussis. PMID- 8717384 TI - Particle-mediated nucleic acid immunization. AB - Nucleic acid immunization involves the direct in vivo administration of antigen encoding plasmid DNA molecules that results in the de novo production of correctly folded microbial antigens at the site of DNA delivery. While this process can lead to the development of neutralizing antibody responses recognizing authentic protein conformations, in vivo antigen production also results in epitope presentation via the MHC class I antigen processing pathway, leading to the elicitation of cytotoxic cellular immune responses. Recent efforts in the authors' laboratories have focused on use of the Accell gene delivery system (gene gun) to achieve the direct, intracellular delivery of small quantities of DNA into cells of the epidermis. The gene gun approach to nucleic acid vaccination capitalizes on the synergistic combination of an effective DNA delivery system and a target tissue that serves as a major immunological inductive site. Experimental gene gun-based nucleic acid vaccines can achieve potent humoral and cytotoxic cellular immune responses in rodent models following immunization with as little as 16 ng of DNA. Equally strong responses have also been elicited in larger animals, such as pigs and monkeys, following epidermal immunization with as little as 2 to 4 micrograms of DNA. PMID- 8717385 TI - Improved method for the removal of endotoxin from DNA. AB - Contaminating endotoxin in solutions used in gene therapy and genetic immunization can result in various deleterious effects both in vitro and in vivo. In order to avoid such complications, attempts were made to characterize the extent of the problem of endotoxin contamination and develop a solution to this problem. After screening for endotoxin in plasmid DNA preparations using the Limulus Amoebocyte Lysate (LAL) assay, nearly half of all samples displayed high endotoxin levels. Therefore, a simple one-step procedure was developed for the removal of endotoxin using a polymyxin B resin. PMID- 8717386 TI - DNA-based immunization against the envelope proteins of the hepatitis B virus. AB - Intramuscular injection in mice of DNA expression vectors encoding the envelope proteins of the hepatitis B virus induced humoral responses specific to several antigenic determinants of the viral envelope. The use of different promoter elements in the plasmid vectors influenced the kinetics and specificity of antibodies produced to the envelope proteins. The first antibodies appeared within 1-2 weeks after injection of DNA and included antibodies of the IgM isotype. Over the following weeks, an IgM-to-IgG class switch occurred, indicating helper T-lymphocyte activity. Peak IgG titers were reached by 4 weeks after a single DNA injection and were maintained for at least 6 months without further DNA injections. The antibodies to the envelope proteins reacted with both group- and subtype-specific antigenic determinants of the HBV surface antigen (HBsAg). The nature of the immune response to the envelope proteins provides indirect evidence that the proteins have adopted a native conformation and have probably been assembled into particles after intramuscular expression from the plasmid vectors. These results indicate that it may be possible to rationally design DNA expression vectors to induce a particular type of immune response for vaccination against hepatitis B or other pathogens. PMID- 8717387 TI - Induction of antibodies against SIV antigens after intramuscular nucleic acid inoculation using complex expression constructs. AB - By studying the infection of rhesus macaques with simian immunodeficiency virus (SIVmac) the potential of nucleic acid immunisation against AIDS can be evaluated. As a first step towards the development of suitable expression constructs, the levels and the durations of expression elicited by the house keeping gene promoters of the murine phospho-glycerate kinase (PGK) gene and rat proto-ras 1Ha, a lentiviral LTR and the CMV-intron A promoter were tested in BALB/c mice intramuscularly inoculated with marker gene constructs encoding luciferase. The expression levels achieved by the CMV-intron A and the lentiviral promoter were comparably high, and also the PGK promoter induced a high level of expression for at least 64 days. Following the inoculation of plasmids comprising single or multiple genes of SIV, the induction of specific antibodies directed against SIV antigens was demonstrated. We previously showed in vitro that int- and nef-defective mutants of SIVmac were able to initiate a limited and self abortive infection of permissive cells in the absence of chromosomal integration of the viral DNA. Intramuscular inoculations in monkeys using int-defective proviral DNA of SIV will show whether an increased immune response may be induced by expression of viruses undergoing a self-limited replication in vivo. PMID- 8717388 TI - Novel approaches to vaccine development against HBV. AB - This review summarizes studies from my laboratory based on two novel approaches towards vaccine development against hepatitis B surface antigen. The first approach deals with the ability of a monoclonal anti-idiotypic antibody to functionally mimic the a determinant of HBsAg. The second approach exploits the strategy of genetically engineering plants so that they can be used as inexpensive alternatives to fermentation systems for the production of candidate vaccine antigens. PMID- 8717389 TI - The use of marker vaccines in eradication of herpesviruses. AB - Marker vaccines are vaccines that allow serological differentiation between infected and vaccinated individuals. This differentiation is based on the absence of one or more microbial proteins in the vaccine that are present in the wild type micro-organism. Consequently, after infection, but not after vaccination, an antibody response against that specific protein(s) can be detected. With a protein-specific antibody test infected individuals can thus be distinguished from vaccinated individuals. Marker vaccines against pseudorabies virus (PRV) and against bovine herpesvirus 1 (BHV1) infections have been developed, along conventional routes and by recombinant DNA technology. These vaccines have been shown to be efficacious in reducing (a) clinical signs after infection, (b) wild type virus replication after infection, and (c) transmission of wild-type virus in the laboratory and in the field. At present, PRV vaccines that lack the gene for the glycoprotein gE are used worldwide in novel eradication programmes. The first phase of such a programme consists of systematic vaccination of pigs on a farm, in a region or an entire country. Experiences in the Netherlands show that it is feasible to eradicate PRV by the intensive use of marker vaccines. Whether, this also holds true for BHV1 is now under investigation. PMID- 8717390 TI - Strategy for producing new foot-and-mouth disease vaccines that display complex epitopes. AB - Widely used inactivated vaccines for foot-and-mouth disease (FMD) induce protective immunity, but vaccine production plants and residual virus in the vaccine itself have been implicated in disease outbreaks. The structure of the FMD virion has been determined, and although much of the surface of the viral particle is produced by complex folding of the three surface-exposed capsid proteins (VP1-3), some surface regions representing important linear epitopes can be mimicked by recombinant proteins or synthetic peptides. Vaccine candidates based on these products stimulate immune responses to foot-and-mouth virus (FMDV), but do not always protect livestock from disease. The basis of protective immunity to FMDV has been explored using genetic engineering to produce antigenic chimeras of the virus. Studies with these chimeras have shown that a strong and protective immune response can be generated in livestock to epitopes outside the sequential epitopes incorporated into previous subunit vaccine candidates. Genetic engineering of the virus has also been used to demonstrate that changes within the sequence encoding an arginine-glycine-aspartic acid (RGD) sequence in VP1 abrogate virus binding to cells in culture, confirming the role of RGD as the receptor binding site. Based on this information, genetically stable viruses which cannot bind to cells have been created by deleting the nucleotides coding the RGD sequence. The receptor binding site-deleted viruses have been shown to be non-infectious in tissue culture, mice, and swine. Cattle vaccinated with these viruses are protected from disease when challenged with virulent FMDV, demonstrating that they could serve as the basis for safer FMD vaccines. PMID- 8717391 TI - Hybrid hepatitis B virus core antigen as a vaccine carrier moiety: I. presentation of foreign epitopes. AB - Hepatitis B virus (HBV) core antigen (HBcAg) is a highly immunogenic subviral particle. Here, we review recent progress in the use of HBcAg as a carrier moiety for heterologous epitopes. To define surface exposed and immunogenic insertion sites for foreign epitopes in HBcAg, peptidic epitopes representing binding sites for virus neutralizing antibodies on the HBV surface antigens were inserted at different positions within HBcAg using genetic engineering in an Escherichia coli expression system (Schodel et al. (1992) J. Virol. 66, 106-114). While fusion to the N-terminus required a linker to become surface accessible, both fusion to the N-terminus and to the C-terminus was compatible with particle assembly and preserved the native antigenicity and immunogenicity of HBcAg. Fusion to an immunodominant internal site of HBcAg reduced the HBcAg immunogenicity and antigenicity and most drastically enhanced the immunogenicity of the inserted foreign epitope. This internal site of HBcAg was used to express circumsporozoite antigen (CS) repeat epitopes of two rodent malaria parasites and of Plasmodium falciparum (Schodel et al. (1994b) J. Exp. Med. 180, 1037-1046 and Schodel et al. (1995a) 95th ASM General Meeting, Washington DC, Abstr. E61). When purified from recombinant Salmonella typhimurium, the hybrid HBcAg-CS proteins were particulate and displayed CS antigenicity as well as reduced HBc antigenicity, as compared to native HBcAg. Immunization of several mouse strains with HBcAg-CS hybrid particles resulted in high titered serum anti-CS antibodies representing all murine IgG isotypes. Immunization of mice with HBcAg or HBcAg-CS particles formulated on alum, complete Freunds or incomplete Freunds adjuvant resulted in equivalent anti-CS and anti-HBc serum antibody titres. The possible influence of carrier-specific immunosuppression was examined and pre-existing immunity to HBcAg did not significantly alter the immunogenicity of hybrid HBcAg particles suggesting that they would be useful carrier moieties for repeated immunizations against multiple haptens or in immune subjects after HBV infection. Examination of T cell recognition of HBcAg-CS particles revealed that HBcAg-specific T cells were universally primed and CS-specific T cells were primed if the insert contained a CS-specific T cell recognition site. This indicates that the internal amino acid position in HBcAg is permissive for the inclusion of heterologous functional T helper as well as B cell epitopes. BALB/c mice immunized with HBcAg CS1 were protected against P. berghei challenge to 90% and 100%, respectively, in two independent experiments. PMID- 8717392 TI - Immunization with recombinant vaccinia viruses expressing structural and part of the nonstructural region of tick-borne encephalitis virus cDNA protect mice against lethal encephalitis. AB - Three recombinant vaccinia viruses containing different fragments of tick-borne encephalitis virus (TBEV) cDNA representing the 5'-noncoding region (5'NCR), all structural and part of the nonstructural regions were constructed. Western blot analysis showed that E and NS1 proteins were expressed and processed correctly in cells infected with recombinant viruses vC-NS1 (coding for C-prM-E-NS1 region) and vC-NS3 (coding for C-prM-E-NS1-NS2A-NS2B-NS3 region). In contrast, in cells infected with recombinant virus v5'C-NS2A (coding for 5'NCR and C-prM-E-NS1-NS2A regions) expression of NS1 protein was greatly reduced and no E protein was detected. Immunization of mice with vC-NS3 induced high levels of TBEV-specific antibodies and protected them against intraperitoneal challenge with 10(7) LD50 of TBEV. The level of protection was very similar to the level of protection achieved by immunization with commercially available inactivated TBEV vaccine. Although the immunization of mice with recombinants vC-NS1 and v5'C-NS2A induced much lower levels of TBEV-specific antibodies, they were still protected against intraperitoneal challenge with 10(4) and 10(3.6) LD50 of TBEV, respectively. The high level of protection against TBEV infection achieved by the immunization of mice with the recombinant vaccinia virus vC-NS3 makes this virus a very attractive candidate for development of a live recombinant vaccine against TBEV. PMID- 8717393 TI - Cell-mediated immune response of macaques immunized with low doses of simian immunodeficiency virus (SIV). AB - Many uninfected people at high risk of HIV infection developed an HIV-specific cellular immune response despite their lack of seroconversion. Therefore, they must have been exposed to HIV without subsequent infection. It has been concluded from these data, that cell-mediated immunity (CMI) rather than humoral immunity might confer protection to HIV infection. Therefore, we tried to induce such a strong CMI in macaques by different immunization strategies. Five or seven animals were immunized with high or low doses of a whole SIV split vaccine. The lower dose of the vaccine provoked a stronger T-helper cell (TH) proliferation than the higher dose, which led to a pronounced humoral immune response. To induce a strong CMI without any specific antibody response, five macaques were inoculated with low doses of infectious SIV. None of these animals seroconverted but each animal developed a SIV-specific TH response. Interestingly, we could neither detect an SIV-specific CTL activity in the animals nor did we find typical TH1- or TH2-like cytokine profiles investigating stimulated bulk-cultures from SIV-exposed animals by RT-PCR. 24 weeks after the first low dose SIV exposure the animals were boosted by a second low dose of SIV followed by a subsequent intravenous challenge with a high dose of SIV 12 weeks later. Unexpectedly, none of the animals was found to be protected against infection and the development of AIDS-like symptoms. PMID- 8717394 TI - Inactivated Marburg virus elicits a nonprotective immune response in Rhesus monkeys. AB - In the present work the kinetics of some indices of immunity (tumor necrosis factor (TNF), interferon (IFN), natural killer cells (NK), lymphocyte proliferation activity, virus-specific antibodies, CD4/CD8 ratio) in response to Marburg virus infection in Macaca mulatta were studied. The different kinetics of immunological parameters for animals which survived Marburg virus infection and for animals which died after infection are shown. A comparison of the indices of IFN, TNF and spontaneous lymphocyte proliferation activity in Macaca mulatta infected with Marburg virus at different stages of life shows the relationship between the increase of these indices and the decrease in the animals' lifespan. Marburg virus immunosuppressive properties were corroborated by studying lymphocyte proliferation activity in response to antigenic stimulation in vitro and the CD4/CD8 index during experimental Marburg virus infection in Macaca mulatta. We conclude that the disease outcome depends on the dynamics of certain immunologic indices such as TNF and IFN. PMID- 8717395 TI - Modification of translational control elements as a new approach to design of attenuated picornavirus strains. AB - The translation machineries of different host cells may exhibit varying requirements for a specific structure of cis-acting control elements in the viral RNA templates. Thus, the appropriately spaced oligopyrimidine/AUG tandem (OAT), a conserved control element in the 5' noncoding region of the picornavirus genomes, is dispensable for the growth of Theiler's murine encephalomyelitis virus (TMEV) in BHK-21 cells, but is essential for the neurovirulence of this virus. Also, the replacement of the cryptic (non-initiator) AUG moiety of the wild-type poliovirus OAT by the initiator AUG affects the viral reproduction in cultured cells only slightly, whereas neurovirulence of the relevant mutants is dramatically suppressed. These observations allow us to propose a rational way to construct novel attenuated viral strains by elimination or severe modification of host specific regulatory regions in their genomes. The relevant genetic rearrangements may be so extensive that the probability of reversion to the virulent phenotype should be negligible. The feasibility of engineering of highly attenuated and genetically stable TMEV and poliovirus variants is illustrated. PMID- 8717396 TI - Design of immunogens as components of a new generation of molecular vaccines. AB - Three new approaches to design effective immunogens are considered. At first, we derived an expression vector from bacteriophage M13 allowing the exposure of short peptides on the virion surface. EIA demonstrates that antibodies against a recombinant phage carrying the antigenic determinant of the HIV-1 gag protein reacted with the 17-kDa core protein of the virus and also with its polyprotein precursor p55 in immunoblotting. In another approach, we chose the hepatitis B core antigen (HBcAg) particle as a vehicle for the presentation of foreign antigenic determinants to the immune system. Chimerical particles of HBcAg containing epitope of the VEE virus were obtained. A vector system for insertion of foreign antigenic determinants and production of both hybrid and wild HBcAg proteins were also obtained. The third approach relies on construction of immunogens from different T- and B-cell epitopes of the HIV-1. We suggested to construct HIV-1 vaccines in a form of the TBI (T- and B-cell epitopes containing Immunogen) with a predetermined tertiary structure, namely, a four-alpha-helix bundle. The gene of the TBI protein consisting of nine HIV-1 epitopes was synthesized and expressed in Escherichia coli cells. Mice immunized with TBI showed humoral and cellular immune responses to HIV-1. Anti-TBI antibodies displayed HIV-1 neutralizing activity. These new approaches offer promise in the development of new effective vaccines. PMID- 8717397 TI - Immunity and vaccine development in Pasteurella multocida infections. AB - The role of LPS in immunity was studied using monoclonal antibodies (MAbs) and active immunisation experiments. A panel of six MAbs produced against Pasteurella multocida serotype B:2 reacted with the LPS of serotypes B:2 and B:5, but not with other serotypes. The MAbs could opsonise P. multocida for phagocytosis by mouse macrophages, but were not bactericidal in the presence of complement. They conferred only partial passive protection in mice. Similar results showing only partial protection were obtained when purified LPS was used to actively immunise mice prior to challenge, suggesting that LPS plays a partial role in immunity to infection. The aroA gene from P. multocida serotypes A:1 and A:3 was cloned and inactivated by insertion of a kanamycin resistance gene. The mutated gene was re introduced onto the chromosome by allelic exchange. The resultant aroA mutants were highly attenuated in a mouse model system, with a 6-log decrease in ID50. Virulence could be restored by complementation with a functional aroA gene. Mice immunised with two doses of the live mutants were protected against lethal challenge with the homologous parental strain, but not against the heterologous strain. P. multocida A:1 and A:3 expressed unique proteins when grown in iron restricted medium. Moreover, the outer membrane (OM) fractions of these cells contained novel proteins of 75 kDa, 85 kDa and 94 kDa molecular mass. Mice were immunised with OM fractions prepared from serotype A:3 grown in iron-restricted (OM Fe-) or iron-replete (OM Fe+) media. When low challenge doses were used, both immunogens protected mice against serotype A:3, but only the OM Fe- fraction protected mice against heterologous challenge with serotype A:1. When higher challenge doses were used, only partial protection was observed. PMID- 8717398 TI - Outer membrane proteins of Pseudomonas aeruginosa as vaccine candidates. AB - We tested the ability of the recombinant outer membrane proteins of Pseudomonas aeruginosa to serve as a protective vaccine against this Gram-negative pathogen in the presence of two main pathophysiological events leading to P. aeruginosa sepsis: (i) systemic infection during immunosuppression; and (ii) bacterial translocation. A hybrid vaccine was cloned which combined the protective epitopes of outer membrane protein F (OprF) and outer membrane protein I (OprI). This vaccine proved to be highly protective against an intraperitoneal challenge with P. aeruginosa in immunosuppressed mice. Oral immunization of mice with recombinant OprI expressing Salmonella dublin, induced s-IgA antibodies in the gut mucosa against OprI. These provided protection against translocation of P. aeruginosa in an immunosuppressed mouse model. To test whether OprI is effective in man, recombinant OprI was purified and used for the immunization of human volunteers. Immunization was tolerated well, and no side effects were observed. Antibody titers against OprI were measured in 90% of the volunteers after immunization. PMID- 8717399 TI - Development of novel vaccines against anthrax in man. AB - It has been shown that antianthrax immunity induced by the novel vaccine proposed has not only antitoxic, but also antispore character. The whole complex of antigens, namely surface spore antigens, surface antigens of cell wall and toxin components is required for the induction of strong and stable immunity against anthrax. The STI-1 vaccine strain with introduced resistance to several antibiotics seems to be promising for prophylaxis and treatment of anthrax in case of emergency, especially if antibiotic pretreatment could be expected. The technology for submerged cultivation of Bacillus anthracis vaccine strain and for the development of an anthrax vaccine to be used in human medicine is proposed on the basis of the conception of the immunogenesis. PMID- 8717400 TI - Bacterial ghosts: non-living candidate vaccines. AB - Expression of cloned PhiX174 gene E in bacteria results in lysis of bacteria. It is unique among phage lysis systems as it introduces a transmembrane tunnel structure through the cell envelope complex of Gram-negative bacteria. The resulting bacterial ghosts have intact envelope structures devoid of cytoplasmic contents. E-mediated lysis has been achieved in a variety of Gram-negative bacteria including Escherichia coli, Salmonella typhimurium, Vibrio cholerae, Klebsiella pneumoniae, and Actinobacillus pleuropneumoniae. Such ghosts, derived from human or animal pathogens, have been proposed as non-living candidate vaccines and represent an alternative to heat or chemically inactivated bacteria. In 'recombinant ghosts', foreign proteins (e.g., viral proteins) are inserted into the inner membrane via specific N-, or C-, or N- and C-terminal anchor sequences prior to lysis. Relevant advantages of (recombinant) bacterial ghosts as immunogens include: (i) inactivation procedures that denature relevant immunogenic determinants are not employed in the production of ghosts used as vaccines or as carriers of relevant antigens; (ii) the recombinant proteins are inserted into a highly immune stimulatory environment; (iii) there is no size limitation of the foreign protein moieties: multiple antigenic determinants can be presented simultaneously; (iv) bacterial ghosts can be produced inexpensively in large quantities; (v) (recombinant) ghosts are stable for long periods of time and do not require the cold chain storage system. Intraperitoneal, subcutaneous or intramuscular applications of recombinant ghosts in experimental animals induced specific humoral and cellular immune responses against bacterial and viral components. Initial aerosol vaccinations of swine with ghosts from Actinobacillus pleuropneumoniae showed that protective immunity can be established by this route of application and that the well-preserved surface structures of ghosts obtained by E-mediated lysis are able to target the mucosal immune system. PMID- 8717401 TI - Induction of protective immunity by aerosol or oral application of candidate vaccines in a dose-controlled pig aerosol infection model. AB - In order to outline basic concepts for the design of a bacterial aerosol infection model, the development of a pig model with Actinobacillus pleuropneumoniae is described. First, reproducibility of aerosol parameters should be maintained by optimizing generating and sampling conditions. Survival rates of the chosen strain must be predictable. Secondly, inhalation conditions for the recipients have to be standardized to enable the determination of deposition sites and the dose administered. Subsequently, dose-response relationship should be evaluated to find a suitable challenge dose. Furthermore, it seems necessary to establish methods to obtain local specimens for determination of the local immune responses. The present study demonstrates that after aerosol challenge pigs were completely protected after inhalation and partially protected after oral application of A. pleuropneumoniae vaccines and describes techniques to administer bacteria in a dose-dependent, viable way. Using the infection model several stages of the disease from acute pleuropneumonia to chronic infection can be induced for research purposes. PMID- 8717402 TI - The potential of Lactobacillus as a carrier for oral immunization: development and preliminary characterization of vector systems for targeted delivery of antigens. AB - Oral administration of lactobacilli evokes mucosal and systemic immune responses against epitopes associated with these organisms (Gerritse et al., 1990, 1991). The adjuvant function of different Lactobacillus species was investigated under the conditions of intraperitoneal (i.p.) injection or oral administration. After i.p. injection of trinitrophenylated chicken gamma-globulin, high DTH responses were observed with Lactobacillus casei and Lactobacillus plantarum, but low responses with Lactobacillus fermentum and Lactobacillus delbrueckii subsp. bulgaricus. In different experimental model systems L. casei and L. plantarum consistently showed significant adjuvanticity. A series of expression and expression-secretion vectors containing the strong constitutive promoter of the L. casei L-ldh gene or the regulatable promoter of the Lactobacillus amylovorus amy gene (Pouwels and Leer, 1995) was used for the intracellular, extracellular and surface-bound expression of an influenza virus antigenic determinant fused to Escherichia coli beta-glucuronidase. Intracellular expression of the fusion protein amounted to 1-2% of total soluble protein. Lactobacilli synthesizing the fusion protein intracellularly evoked an oral immune response after subcutaneous priming. PMID- 8717403 TI - Attenuated Salmonella as live oral vaccines against typhoid fever and as live vectors. AB - Attenuated Salmonella typhi vaccine strain CVD 908, which harbors deletion mutations in aroC and aroD, has been shown to be well-tolerated and highly immunogenic, eliciting impressive serum antibody, mucosal IgA and cell-mediated immune responses. A further derivative prepared by introducing a deletion in htrA (which encodes a heat-shock protein that also has activity as a serine protease in CVD 908 (Chatfield et al., unpublished data) resulted in CVD 908-htrA. In phase 1 clinical trials, CVD 908-htrA appears very attractive as a live oral vaccine candidate. Both CVD 908 and CVD 908-htrA are useful as live vector vaccines to deliver foreign antigens to the immune system. Conditions that enhance the expression and immunogenicity of foreign antigens carried by CVD 908 and CVD 908-htrA are being investigated. PMID- 8717404 TI - Mixed population approach for vaccination with live recombinant Salmonella strains. AB - Attenuated strains of enteropathogenic species, such as Salmonella, represent useful carries for the delivery of heterologous recombinant antigens to the immune system. A frequently encountered obstacle, however, is the negative influence of high-level antigen production on the stability of carrier strains and the maintenance of their specific properties concerning tissue colonization and viability during infection. To solve this problem we have established an expression system based on genetic variation. This generates two sub-populations of a recombinant vaccine strain, i.e., one consisting of viable cells which maintain all characteristics of the native carrier strain and generate a second population of cells producing antigen(s) of interest at a very high level. This novel expression system offers unique applications and advantages over common live recombinant vaccine approaches. PMID- 8717405 TI - Optimization of live oral Salmonella-HIV-1 vaccine vectors for the induction of HIV-specific mucosal and systemic immune responses. AB - Recent evidence suggests that live oral Salmonella-HIV vaccine vectors have the potential to elicit HIV-specific T cell-mediated immunity in both the mucosal and systemic compartments. We are using the mouse-typhoid model to identify Salmonella::HIV vaccine vector constructs that elicit HIV-specific mucosal and systemic immune responses. Oral immunization of mice with a Salmonella strain that expresses recombinant gp120 (rgp120) in the cytoplasm of the vector elicits a modest gp120-specific T cell proliferation response in the spleen. However, such Salmonella constructs did not stimulate the development of gp120-specific serum IgG or cytotoxic T lymphocytes (CTLs). Interestingly, the majority of cytoplasmically-expressed rgp120 forms inclusion bodies in Salmonella. We believe that in this form rgp120 is highly susceptible to protease degradation by the vector. As such, cytoplasmic rgp120 may not persist in the host after vaccination, resulting in the modest immunogenicity of rgp120 in these constructs. To circumvent this problem we constructed Salmonella strains that express rgp120 on the surface of the vector. Preliminary data suggest that surface-expressed rgp120 is significantly more immunogenic in both the mucosal and systemic compartments than cytoplasmic rgp120. These results, therefore, support the proposal that Salmonella vectors will be a safe and inexpensive means for delivery of HIV antigens to, and the elicitation of HIV-specific T cells in, the mucosal and systemic compartments. PMID- 8717406 TI - Mucosal immunity: regulation by helper T cells and a novel method for detection. AB - The mechanisms which regulate mucosal IgA responses to orally administered protein vaccines are not yet fully elucidated. We have used two delivery systems, soluble tetanus toxoid (TT) with the mucosal adjuvant cholera toxin (CT) and recombinant Salmonella expressing Tox C, a fragment of TT, to assess the nature of CD4+ T helper (Th) cells and derived cytokines which support mucosal IgA responses in both normal and cytokine knockout (interferon gamma knockout; IFN gamma-/- and IL-4-/-) mice. Our results provide important new information regarding Th cell and cytokine regulation of mucosal IgA responses. Whereas TT coadministered with CT induces predominant TT-specific Th2-type responses, rSalmonella delivery of Tox C induced dominant Th1-type responses along with synthesis of the Th2-cytokine IL-10. Both vaccine regimen elicited high levels of mucosal S-IgA and IL-6 production by macrophages. Further oral immunization of IFN-gamma-/- and IL-4-/- mice with rSalmonella Tox C also induced macrophage derived IL-6 and Th2-derived IL-10 as well as S-IgA responses, suggesting that IFN-gamma from Th1-type cells as well as traditional Th2 cells producing IL-4 and IL-5 are not essential for mucosal IgA responses. Rather, induction of second level Th2 cells producing IL-10 together with high levels of IL-6 from other cell sources may be sufficient for mucosal IgA responses in the absence of traditional Th2 cells. These studies were facilitated by the development of a sensitive new luminometry assay which allowed detection of cytokines and cell surface molecules which are below the levels of detection by current solid phase assays. PMID- 8717407 TI - Peripheral blood antibody-secreting cells in the evaluation of the immune response to an oral vaccine. AB - Specific antibody-secreting cells (ASC) appear in the blood as a response to oral vaccination in humans. Based on information from animal experiments, these cells are believed to be migrating to the mucosa. This review summarizes a series of studies aimed at a detailed characterization of the ASC response to a prototype oral vaccine Salmonella typhi Ty21a, with respect to its kinetics, Ig-class distribution, antigen specificity, influence of the administration route and nature of the antigen, and the corresponding antibody responses in serum. Different vaccine formulations as well as dosage schedules are compared, and the response to booster immunization is described. The response manifested by ASC in blood is shown to be independent from serum antibody responses. Moreover, it is shown to parallel with the results obtained for protection in field trials. Finally, some data on the homing receptor expression of these cells are presented, giving further evidence for the mucosal homing of these cells. The ASC assay offers a practical means for assessing immune response to oral vaccines in humans. It can be used as a laboratory parameter correlated with protection conferred by an oral typhoid vaccine. It can even be applied to measure active mucosal immunity, i.e., protective immunity by showing the relative reduction of the ASC response to an oral dose of live vaccine. PMID- 8717408 TI - Toward selective elicitation of TH1-controlled vaccination responses: vaccine applications of bacterial surface layer proteins. AB - Bacterial surface layer proteins have been utilized as combined vaccine carrier/adjuvants and offer a number of advantages in these applications. The crystalline protein arrays contain functional groups in precisely defined orientations for coupling of haptens. Conventional applications of S-layer vaccines do not cause observable trauma or side effects. Depending on the nature of the S-layer preparations, antigenic conjugates will induce immune responses of a predominantly cellular or predominantly humoral nature. Immune responses to S layer-hapten conjugates are also observed following oral/nasal application. In the present contribution, the status of investigations with S-layer conjugates in three main immunological projects is reviewed. In a project aimed at immunotherapy of cancer, conjugates of S-layer with small, tumor-associated oligosaccharides have been found to elicit hapten-specific DTH responses. An enlarged program of chemical synthesis has now been initiated to prepare a complete set of mucin-derived, tumor-associated oligosaccharides and their chemically modified analogues for elicitation of cell-mediated immune responses to certain tumors in humans. In another application, oligosaccharides derived from capsules of Streptococcus pneumoniae type 8 have been linked to S-layer proteins and have been found to elicit protective antibody responses in animals. Most recently, allergen S-layer conjugates have been prepared with the intention to suppress the TH2-directed, IgE-mediated allergic responses to Bet nu 1, the major allergen of birch pollen. In the former two applications, the S-layer vaccine technology appears to offer the versatility needed to direct vaccination responses toward predominant control by TH1 or TH2 lymphocytes to meet the different therapeutic or prophylactic requirements in each case. In the third application, work has progressed to a preliminary stage only. PMID- 8717409 TI - Intracellular, genetic or congenital immunisation--transgenic approaches to increase disease resistance of farm animals. AB - Novel approaches to modify disease resistance or susceptibility in livestock are justified not only by economical reasons and with respect to animal welfare but also by recent advancements in molecular genetics. The control or elimination of infectious pathogens in farm animals is historically achieved by the use of vaccines and drugs and by quarantine safeguards and eradication. Currently, research on the improvement of disease resistance based on nucleic acid technology focuses on two main issues: additive gene transfer and the development of nucleic acid vaccines. The strategies aim at the stable or transient expression of components known to influence non-specific or specific host defence mechanisms against infectious pathogens. Thus, candidates for gene transfer experiments include all genes inducing or conferring innate and acquired immunity as well as specific disease resistance genes. Referring to the site and mode of action and the source of the effective agent the strategies are termed 'intracellular', 'genetic' and 'congenital' immunisation. The targeted disruption (deletive gene transfer) of disease susceptibility genes awaits the establishment of totipotential embryonic cell lineages in farm animals. The cytokine network regulates cellular viability, growth and differentiation in physiological and pathophysiological states. The identification of the JAK-STAT pathway used by many cytokines for their intracellular signal propagation has provided not only new target molecules for modulating the immune response but will also permit the further elucidation of host-pathogen interactions and resistance mechanisms. PMID- 8717411 TI - Biochemical and immunological identification and enrichment of poly(A) polymerase from human thymus. AB - Human thymus poly(A) polymerase (EC 2.7.7.19) activity has been investigated using poly(A) and oligo(A) as initiators. All obtained fractions reveal more than one polypeptide as detected by immunoblotting after SDS-PAGE. In addition to the homogeneously purified (Tsiapalis et al., J Biol Chem 250: 4486-4496, 1975 and Wahle, J Biol Chem 266: 3131-3139, 1991), about 60 kDa polypeptide, a larger polypeptide, about 80 kDa, that comigrates in the region of poly(A) polymerase activity was detected, enriched and partially characterized; it appears having similar size with bovine poly(A) polymerase cloned in E. coli. Polyclonal antiserum produced against recombinant bovine poly(A) polymerase reacts more efficiently with the about 80 kDa polypeptide upon immunoblotting, and can precipitate the poly(A) polymerase activity. This enzyme form, from human tissue, is novel in terms of size and may reflect intact or physiological form of poly(A) polymerase in human thymus, and supports and substantiates recent reports on the enzyme from other sources. PMID- 8717410 TI - Decrease in the glycosaminoglycan content in the skin of diabetic rats. The role of IGF-I, IGF-binding proteins and proteolytic activity. AB - The results of our previous studies demonstrated that acute streptozotocin induced diabetes in rats evoked a decrease in skin collagen content with little effect on glycosaminoglycans (GAG) content. In our present study we employed the model of chronic diabetes in order to check its effect on skin GAG content. It was found that the skin of diabetic rats showed a significant decrease in almost all the investigated GAGs by 50-70%. The decrease in heparan sulfate content was slight and statistically insignificant. We sought to determine whether the insulin-like growth factor-I (IGF-I) and IGF-binding proteins (IGF-BPs) levels are altered in animals with experimental diabetes and might contribute to the decrease in tissue GAG content. Circulating IGF-I level was found to be reduced in animals with diabetes and significant changes in serum IGF-BPs were observed. The amount of high molecular weight binding proteins (HMW-BPs) was decreased and the content of low molecular weight binding proteins (LMW-BPs), known as IGF-I inactivating substances, markedly increased. Furthermore, diabetic rats demonstrated an increase of skin proteolytic activity. We conclude that the decrease of GAG content in the skin of diabetic rats is a result of three co existing phenomena: decreased circulating IGF-I level, increased plasma content of LMW-BPs and increased proteolytic activity of the skin. PMID- 8717412 TI - Role of thyroid hormones in renal tubule acidification. AB - Renal tubule acidification was studied in thyroparathyroidectomized rats which had the parathyroids reimplanted into cervical muscle tissue, by stopped-flow microperfusion using ion-exchange resin microelectrodes. Hypothyroid rats had decreased rates of proximal and late distal bicarbonate reabsorption. This reduction occurred in the absence of changes in pH gradients, and was due mostly to decreases in acidification half-times, that is, of the rate of bicarbonate exit from the tubule lumen. H+ back-flux from the lumen measured during luminal perfusion with solutions at pH 6 (below stationary pH) was decreased in proximal tubule of hypothyroid rats, showing that the acidification defect was not due to an increased H+ shunt across the epithelium. These data indicate that in hypothyroid rats the proximal tubule luminal density of Na+/H+ exchangers or their turnover is decreased in the absence of alterations in the driving force (H+ and Na+ gradients across the luminal membrane) for H+ secretion. The effect observed in distal tubule may be due to action on Na+/H+ exchangers that are present also on this site, or to an impairment of the action of other H+ transporters such as H(+)-ATPases, including the provision of energy for them. 9 PMID- 8717413 TI - Oxygen-dependent leishmanicidal activity of stimulated macrophages. AB - Peritoneal macrophages pretreated with different stimulants were analysed and compared with their respective controls for their ability to kill intracellular pathogenic L. donovani, (MHOM/IN/1983/AG83) an isolate from Indian subcontinent. Stimulation of macrophages by zymosan showed a higher microbicidal activity as compared to that by PMA. A correlation between microbicidal activity of the macrophages and the parameters related to respiratory burst activity such as liberation of O2-, production of H2O2 and consumption of O2 was sought. All the parameters showed a decrease in case of infected macrophages in comparison to those of the non-infected ones. Thus, it is possible that the impairment of macrophage activation by intracellular Leishmania contributes to their survival in the toxic environment of the host. PMID- 8717414 TI - Pharmacology of endothelins: vascular preparations for studying ETA and ETB receptors. AB - Three rabbit vessels, the carotid and pulmonary arteries and the jugular vein were investigated to identify vascular monoreceptor systems (either ETA or ETB) to be used in structure-activity studies on endothelins and their antagonists. The RbCA has been found to behave as a monoreceptor ETA preparation, since it shows much greater sensitivity to ET-1 than to ET-3 and is insensitive to IRL 1620. The contractile response of the RbCA to ET-1 is reduced in the presence of BQ-123 but is not influenced by BQ-788. The RbPA behaves as a pure ETB system when stimulated with the ETB selective agonist IRL 1620. The contractile effect of IRL 1620 is reduced in the presence of BQ-788 but is not influenced by BQ-123. The RbJV responds to ET-1 and to IRL 1620 with contractions that are reduced by both BQ-123 and BQ-788, respectively. THe RbJV appears to be a mixed ETA and ETB system in which the two functional sites play an equivalent role in the stimulatory contractile response. Thus, contractile ETA and ETB receptors have been found in arterial and venous vessels of the rabbit and some of these vessels provide sensitive and selective (either ETA or ETB) preparations that appear to be adequate for pharmacological studies on ET receptor agonists or antagonists. PMID- 8717415 TI - tBHP induced in vitro swelling of rat liver mitochondria. AB - Tert-butyl hydroperoxide induced swelling of freshly isolated rat liver mitochondria was inhibited by butylated hydroxytoluene, butylated hydroxyanisole and alpha-tocopherol by acting at the initial phase. EDTA was more effective than EGTA in reducing the initial swelling and so were desferal and bipyrridyl. Spermine, an allosteric activator of calcium uptake, enhanced swelling whereas lanthanum and ruthenium red, the Ca2+ uniport blockers, reduced it. Inhibition of phospholipase A2 by dibucaine and Ca2+ activated proteases by antipain and leupeptin also reduced t-BHP induced swelling. The data indicate that peroxidative mitochondrial swelling involves an iron mediated initial rapid phase and a subsequent calcium dependent propagation phase. PMID- 8717416 TI - Partial purification of an IGF-II receptor/binding protein from the erythroleukemia cell line K562. AB - We previously reported that insulin-like growth factor II (IGF-II) stimulated clonal growth of an erythroleukemia cell line, K562, in semi-solid agar, an effect not mimicked by insulin-like growth factor I (IGF-I), as IGF-I receptors are generally not expressed in this cell line. Affinity crosslinking of intact K562 cells with 125I-IGF-II revealed that the labeled hormone predominantly bound to a protein with a molecular weight of approximately 75 K. We report here the partial purification of the 75 K IGF-II binding protein from K562 cells. Triton X 100-solubilized K562 cells were subjected to Sephacryl-400, followed by Sephacryl 200 chromatography. Fractions of interest were collected and applied to a Sepharose-IGF-II column or an immuno-affinity column. The immuno-affinity column was prepared using an antiserum against placental membrane-derived material eluted from the Sephacryl-400 column in the elution volume, corresponding to the IGF-II binding protein from K562 cells. An affi-gel 10 affinity column, prepared with a protein A purified IgG fraction of this antiserum (antibody-29), retarded proteins showing binding specificity for IGF-II, with apparent molecular weights of 76 K, 87 K, and 70 K under reducing conditions. These protein bands were similar to the proteins retarded in the IGF-II affinity column, when evaluated by affinity crosslinking and SDS-PAGE. Fractionation of the purified material from the antibody-29 affinity column on Superose 12 revealed 6 protein peaks. Affinity crosslinking of the peak fractions from FPLC resulted in single bands with a molecular weight of 75 K under reducing conditions with variable specificity for IGF-II. PMID- 8717417 TI - Evaluation of hydroxyl radical-scavenging property of garlic. AB - Garlic has been reported to provide protection against hypercholesterolemic atherosclerosis and ischemia-reperfusion-induced arrhythmias and infarction. Oxygen free radicals (OFRs) have been implicated as causative factors in these diseases and antioxidants have been shown to be effective against these conditions. The effectiveness of garlic in these disease states could be due to its ability to scavenge OFRs. However, the OFR-scavenging activity of garlic is not known. Also it is not known if its activity is affected by cooking. We therefore investigated, using high pressure liquid chromatography, the ability of garlic extract (heated or unheated) to scavenge exogenously generated hydroxyl radical (.OH). .OH was generated by photolysis of H2O2 (1.2-10 mumoles/ml) with ultraviolet (UV) light and was trapped with salicylic acid (500 nmoles/ml). H2O2 produced .OH in a concentration-dependent manner as estimated by .OH adduct products 2,3-dihydroxybenzoic acid (DHBA) and 2,5-DHBA. Garlic extract (5-100 microliters/ml) produced an inhibition (30-100%) of 2,3-DHBA and 2,5-DHBA generated by photolysis of H2O2 (5.00 pmoles/ml) in a concentration-dependent manner. Its activity is reduced by 10% approximately when heated to 100 degrees C for 20, 40 or 60 min. The extent of reduction in activity was similar for the three heating periods. Garlic extract prevented the .OH-induced formation of malondialdehyde in the rabbit liver homogenate in a concentration-dependent manner. It alone did not affect the MDA levels in the absence of .OH. These results indicate that garlic extract is a powerful scavenger of .OH and that heating reduces its activity slightly. PMID- 8717418 TI - Torsionally-strained DNA and intermolecular purine-purine-pyrimidine triple-helix formation. AB - A potentially powerful pharmacological approach to modulating the expression of specific, disease-related genes involves the inhibition of transcription factor binding to promoter or enhancer elements through oligonucleotide-mediated triple helix formation. In vivo, the typical target for intermolecular triplex formation would most likely be torsionally-strained rather than relaxed duplex DNA. To determine the effects of strained DNA on triplex formation, we investigated the interactions between a G/T-rich oligonucleotide and both supercoiled and relaxed plasmid DNA using a restriction endonuclease protection assay. Both the kinetics of formation and dissociation of purine-motif triplexes were unaffected by the conformational state of the duplex DNA. Similarly, the topological state of the plasmid targets was not affected by triplex formation. Taken together, these observations suggest that stable intermolecular triplexes can form in vivo under conditions of moderate torsional strain. PMID- 8717419 TI - Plasma alpha- and gamma-tocopherol have different pattern during normal human pregnancy. AB - Plasma alpha- and gamma-tocopherol were monitored in pregnant women throughout healthy gestational periods and after delivery and were compared with that of non pregnant women. The mean plasma alpha-tocopherol and gamma-tocopherol concentrations in non pregnant Saudi women (15.2 +/- 1.3 and 1.8 +/- 0.2 mumol/l respectively) were found within normal range. The maternal plasma alpha tocopherol level steadily increased reaching maximum level (19.1 +/- 1.6 mumol/l) at late gestation and then gradually decreased after delivery. On the contrary, the optimum level of gamma-tocopherol (2.1 +/- 0.2 mumol/l) was at mid gestation, followed by a progressive decrease until one month after delivery (1.5 +/- 0.1 mumol/l). This study shows that the maternal plasma alpha- and gamma-tocopherol have different profiles that may be attributed to their different responses to the changes in maternal lipids during pregnancy. PMID- 8717420 TI - Vitamin A supplementation inhibits chemiluminescence and lipid peroxidation in isolated rat liver microsomes and mitochondria. AB - In the present study we investigated if administration of vitamin A could protect rat liver microsomes and mitochondria from in vitro peroxidation. Appreciable decrease of chemiluminescence and lipid peroxidation was measured in microsomal membranes from rats receiving vitamin A, with respect to control animals. In membranes derived from control animals, the fatty acid composition was profoundly modified when subjected to in vitro peroxidation mediated by ascorbate-Fe++, with a considerable decrease of 20:4 n6 and 22:6 n3 in mitochondria and 18:2 n6 and 20:4 n6 in microsomes. As a consequence the peroxidizability index, a parameter based on the maximal rate of oxidation of specific fatty acids was higher in supplemented animals than in control group when both kind of membranes were analyzed. These changes were less pronounced in membranes derived from rats receiving vitamin A. These results are in agreement with previous results that indicated that vitamin A may act as an antioxidant protecting membranes from deleterious effects. PMID- 8717421 TI - Fructose-1,6-bisphosphate as a metabolic substrate in hog ileum smooth muscle during hypoxia. AB - Exogenously applied fructose-1,6-bisphosphate has been reported to be effective in preventing some damage to the small intestine during ischemia. To determine whether exogenously applied fructose-1,6-bisphosphate protects ileum smooth muscle from damage from hypoxia and from reoxygenation, we examined the effect of fructose-1,6-bisphosphate on the ability of hog ileum smooth muscle to maintain isometric force during hypoxia and to generate isometric force after reoxygenation in the presence of 5 mM glucose. After 180 min of hypoxia, tissues incubated with 20 mM fructose-1,6-bisphosphate maintained significantly greater levels of isometric force than tissues incubated in the absence of exogenous substrate (23% of pre-hypoxia force compared to 16%). During the first contraction following reoxygenation there was a significantly greater force generation in tissues incubated with 20 mM fructose-1,6-bisphosphate during the hypoxia period compared to tissues with no exogenous substrate included during the hypoxia period (29% of pre-hypoxia force compared to 19%). However, glucose always was a better metabolic substrate compared to fructose-1,6-bisphosphate under all experimental conditions. The presence of fructose-1,6-bisphosphate during hypoxia likely improved tissue function by fructose-1,6-bisphosphate entering the cells and acting as a glycolytic intermediate, since during a 120 min period of hypoxia, unmounted ileum smooth muscle metabolized 1,6-13C-fructose 1,6-bisphosphate to 3-13C-lactate. This conversion of 1,6-13C-fructose-1,6 bisphosphate to 3-13C-lactate was inhibited by the addition of 1 mM iodoacetic acid, a glycolytic inhibitor. We conclude that exogenously provided fructose-1,6 bisphosphate does provide modest protection of ileum smooth muscle from hypoxic damage by functioning as a glycolytic intermediate and improving the cellular energy state. PMID- 8717422 TI - The possible relevance of autoxidative glycosylation in glucose mediated alterations of proteins: an in vitro study on myofibrillar proteins. AB - The present work was carried out to examine the role of glycation and transition metal catalysed autoxidation of sugars in glucose-mediated alterations of myofibrillar proteins. Myofibrils were prepared from rat skeletal muscle and incubated with 1) sugar alone 2) sugar and micromolar concentrations of transition metals (Cu2+ or Fe3+) 3) transition metals alone and the control remained without sugar or transition metals. A significant increase in extent of glycation and decrease in ATPase activity of myofibrils incubated under autoxidative conditions were observed over the other three incubations. Reducing agent 2-mercaptoethanol was highly effective in preventing the alterations induced by glucoxidation, compared to EDTA and aminoguanidine, suggesting the involvement of thiol group oxidation in the reduced function of the protein. Free radical scavengers like catalase, benzoic acid and mannitol were also effective in preventing glucose mediated alterations. Although a high concentration of glucose alone has an insignificant effect on myofibrils in vitro, the results from the present work suggest that glucose in combination with transition metals could lead to functional alterations of myofibrils, and this process by generating free radicals may contribute to the overall complications of diabetes and aging. PMID- 8717423 TI - Ethanol-dependent oxygen consumption and acetaldehyde formation during vanadyl oxidation by H2O2. AB - Sequential addition of vanadyl sulfate to a phosphate-buffered solution of H2O2 released oxygen only after the second batch of vanadyl. Ethanol added to such reaction mixtures progressively decreased oxygen release and increased oxygen consumption during oxidation of vanadyl by H2O2. Inclusion of ethanol after any of the three batches of vanadyl resulted in varying amounts of oxygen consumption, a property also shared by other alcohols (methanol, propanol and octanol). On increasing the concentration of ethanol, vanadyl sulfate or H2O2, both oxygen consumption and acetaldehyde formation increased progressively. Formation of acetaldehyde decreased with increase in the ratio of vanadyl:H2O2 above 2:1 and was undetectable with ethanol at 0.1 mM. The reaction mixture which was acidic in the absence of phosphate buffer (pH 7.0), released oxygen immediately after the first addition of vanadyl and also in presence of ethanol soon after initial rapid consumption of oxygen, with no accompanying acetaldehyde formation. The results underscore the importance of some vanadium complexes formed during vanadyl oxidation in the accompanying oxygen-transfer reactions. PMID- 8717425 TI - Implication of the nucleus in excitation contraction coupling of heart cells. AB - In the present study, Fluo-3 Ca2+ measurement and confocal microscopy techniques were used in order to localize cytosolic []c and nuclear []n free Ca2+ distribution in resting and spontaneously contracting single heart cells from 10 day-old chick embryos. In resting single cells, the concentration of Ca2+ in the cytoplasm was lower than that in the nucleus. Increasing cytosolic free Ca2+ from 100-1600 nM gradually increased [Ca2+]n with a maximum capacity near 1200 nM. Results from Fura-2 microfluorometry and Fluo-3 confocal microscopy suggest a potential cross talk between the increase of cytosolic free Ca2+ and the uptake and release of Ca2+ by the nucleus during spontaneous contraction of single myocytes. Calcium waves in spontaneously contracting cells were found to spread from one cell to the next with the nucleus acting as a fluorescent beacon in which Ca2+ levels remained elevated for several milliseconds even after cytosolic Ca2+ had returned to near basal values. These results strongly suggest that the nucleus plays a negative and positive feedback role in controlling cytosolic free Ca2+ concentration during excitation-contraction coupling in heart cells. PMID- 8717424 TI - Influence of thyroid hormones on the human ATP synthase beta-subunit gene promoter. AB - The action of thyroid hormones on the expression of the mitochondrial ATP synthase beta-subunit gene (ATPsyn beta) is controversial. We detected a binding site for the thyroid hormone receptor between -366 and -380 in the human ATPsyn beta gene by DNase I footprint analysis and band-shift assays. However, expression vectors in which the chloramphenicol acetyl transferase (CAT) reporter gene is driven by the 5' upstream region of ATPsyn beta gene were unresponsive to T3 when transiently transfected to HepG2 or GH4C1 cells. CAT constructs driven by the rat phosphoenolpyruvate carboxykinase (PEPCK) or the growth hormone (GH) promoters were stimulated several fold by T3 in parallel experiments. It is proposed that the biological effects of thyroid hormones on the ATPsyn beta expression occur through indirect mechanisms. PMID- 8717426 TI - The role of magnesium in regulating CCK-8-evoked secretory responses in the exocrine rat pancreas. AB - This study investigates the effect of magnesium (Mg2+) on the secretory responses and the mobilization of calcium (Ca2+) and Mg2+ evoked by cholecystokinin octapeptide (CCK-8) in the exocrine rat pancreas. In the isolated intact perfused pancreas CCK-8 (10(-10) M) produced marked increases in juice flow and total protein output in zero and normal (1.1 mM) extracellular Mg2+ [Mg2+]o compared to a much reduced secretory response in elevated (5 mM and 10 mM) [Mg2+]o. Similar effects of perturbation of [Mg2+]o on amylase secretion and 45 Ca2+ uptake (influx) were obtained in isolated pancreatic segments. In pancreatic acinar cells loaded with the fluorescent bioprobe fura-2 acetomethylester (AM), CCK-8 evoked marked increases in cytosolic free Ca2+ concentration [Ca2+]i in zero and normal [Mg2+]o compared to a much reduced response in elevated [Mg2+]o. Pretreatment of acinar cells with either dibutyryl cyclic AMP (DB2 cAMP) or forskolin had no effect on the CCK-8 induced changes in [Ca2+]i. In magfura-2 loaded acinar cells CCK-8 (10(-8) M) stimulated an initial transient rise in intracellular free Mg2+ concentration [Mg2+]i followed by a more prolonged and sustained decrease. This response was abolished when sodium (Na+) was replaced with N-methyl-D-glucamine (NMDG). Incubation of acinar cells with 10 mM Mg2+ resulted in an elevation in [Mg2+]i. Upon stimulation with CCK-8, [Mg2+]i decreased only slightly compared with the response obtained in normal [Mg2+]o. CCK-8 caused a net efflux of Mg2+ in pancreatic segments; this effect was abolished when extracellular sodium [Na+]o was replaced with either NMDG or choline. The results indicate that Mg2+ can regulate CCK-8-evoked secretory responses in the exocrine pancreas possibly via Ca2+ mobilization. Moreover, the movement of Mg2+ in pancreatic acinar cells is dependent upon extracellular Na+. PMID- 8717427 TI - Stimulation of human cheek cell Na+/H+ antiporter activity by saliva and salivary electrolytes: amplification by nigericin. AB - Proton-dependent, ethylisopropylamiloride (EIPA)-sensitive Na+ uptake (Na+/H+ antiporter) studies were performed to examine if saliva, and ionophores which alter cellular electrolyte balance, could influence the activity of the cheek cell Na+/H+ antiporter. Using the standard conditions of 1 mmol/l Na+, and a 65:1 (inside:outside) proton gradient in the assay, the uniport ionophores valinomycin (K+) and gramicidin (Na+) increased EIPA-sensitive Na+ uptake by 177% (p < 0.01) and 227% (p < 0.01), respectively. The dual antiporter ionophore nigericin (K(+) H+) increased EIPA-sensitive Na+ uptake by 654% (p < 0.01), with maximal Na+ uptake achieved by 1 min and at an ionophore concentration of 50 mumol/l, with an EC50 value 6.4 mumol/l. Pre-incubation of cheek cells with saliva or the low molecular weight (MW) components of saliva (saliva activating factors, SAF) for 2 h at 37 degrees C, also significantly stimulated EIPA-sensitive Na+ uptake. This stimulation could be mimicked by pre-incubation with 25 mmol/l KCl or K(+) phosphate buffer. Pre-incubating cheek cells with SAF and the inclusion of 20 mumol/l nigericin in the assay, produced maximum EIPA-sensitive Na+ uptake. After pre-incubation with water, 25 mmol/l K(+)-phosphate or SAF, with nigericin in all assays, the initial rate of proton-gradient dependent, EIPA-sensitive Na+ uptake was saturable with respect to external Na+, with Km values of 0.9, 1.7, and 1.8 mmol/l, and Vmax values of 13.4, 25.8, and 31.1 nmol/mg protein/30 sec, respectively. With 20 mumol/l nigericin in the assay, Na+ uptake was inhibited by either increasing the [K+]o in the assay, with an ID50 of 3 mmol/l. These results indicate that nigericin can facilitate K+i exchange for H+o and the attending re acidification of the cheek cell amplifies 22Na+ uptake via the Na+/H+ antiporter. The degree of stimulation of proton-dependent, EIPA-sensitive Na+ uptake is therefore dependent, in part, on the intracellular [K+]i. PMID- 8717428 TI - Non-proline-dependent protein kinases phosphorylate several sites found in tau from Alzheimer disease brain. AB - Of 21 phosphorylation sites identified in PHF-tau 11 are on ser/thr-X motifs and are probably phosphorylated by non-proline-dependent protein kinases (non-PDPKs). The identities of the non-PDPKs and how they interact to hyperphosphorylate PHF tau are still unclear. In a previous study we have shown that the rate of phosphorylation of human tau 39 by a PDPK (GSK-3) was increased several fold if tau were first prephosphorylated by non-PDPKs (Singh et al., FEBS Lett 358: 267 272, 1995). In this study we have examined how the specificity of a non-PDPK for different sites on human tau 39 is modulated when tau is prephosphorylated by other non-PDPKs (A-kinase, C-kinase, CK-1, CaM kinase II) as well as a PDPK (GSK 3). We found that the rate of phosphorylation of tau 39 by a non-PDPK can be stimulated if tau were first prephosphorylated by other non-PDPKs. Of the four non-PDPKs only CK-1 can phosphorylate sites (thr 231, ser 396, ser 404) known to be present in PHF-tau. Further, these sites were phosphorylated more rapidly and to a greater extent by CK-1 if tau 39 were first prephosphorylated by A-kinase, CaM kinase II or GSK-3. These results suggest that the site specificities of the non-PDPKs that participate in PHF-tau hyperphosphorylation can be modulated at the substrate level by the phosphorylation state of tau. PMID- 8717430 TI - Alterations in certain lysosomal glycohydrolases and cathepsins in rats on dexamethasone administration. AB - Glucocorticoids have been used in the treatment of a number of diseases where immunological intolerance plays a predominant role. Since immunological intolerance points to the involvement of lysosomal enzymes and glucocorticoids are known to affect their activities, we have attempted to study the effect of these steroids on cardiac and renal enzymes. Dexamethasone, a glucocorticoid, is administered subcutaneously to male Wistar rats at a dosage of 2.5 mg/kg/week on alternate days for two weeks. After withdrawing the steroid, the animals are monitored for one week to oversee the recovery process. Total and free activities of glycohydrolases and cathepsins in serum, heart and kidney are assayed on the days 4, 8, 12, 16 of dexamethasone administration and also on days 4 and 8 following discontinuation of the steroid. During dexamethasone administration, a significant decrease in both the free and total activities of beta-glucuronidase, beta-N-acetyl glucosaminidase, beta-galactosidase, alpha-galactosidase, alpha mannosidase, cathepsin B and cathepsin D are observed in heart and kidney, but the enzyme levels are shown to increase in serum. On withdrawal of the steroid, the activities of beta-glucuronidase, beta-N-acetyl glucosaminidase, beta galactosidase are found to be increased in heart and kidney, whereas, the activity of alpha-mannosidase remains within normal values. Thus, it could be seen that dexamethasone alters the pattern of glycohydrolases and cathepsins, which are involved in protein degradation. PMID- 8717429 TI - The association of phosphorylase kinase with membranes of rat liver smooth endoplasmic reticulum. AB - Upon fractionation of a post mitochondrial supernatant from rat liver, phosphorylase kinase activity was largely recovered in the cytosol and the smooth endoplasmic reticulum (SER) fraction. The presence of phosphorylase kinase in SER vesicles was not due to an interaction of the enzyme with glycogen particles, since previous elimination of SER glycogen either by 48 h animal starvation or by treatment of the membrane fraction with alpha-amylase did not significantly alter phosphorylase kinase activity content. Washing of the initial pellet of SER fraction (crude SER) by dilution and recentrifugation, released in the supernatant an amount of phosphorylase kinase activity, which is dependent on: i) the degree of dilution, ii) the number of washes, iii) the ionic strength of the washing solution and iii) the presence or absence of Ca2+. Crude SER-associated phosphorylase kinase was marginally affected by increased concentrations of antibody against rabbit skeletal muscle holoenzyme which nevertheless drastically inhibited cytosolic enzyme activity, while it showed a higher resistance to partial proteolysis and a different Western blotting profile with anti phosphorylase kinase when compared with the soluble kinase. A small but significant fraction of SER phosphorylase kinase was strongly associated with the microsomal fraction being partly extractable only in presence of detergents. This membrane-bound enzyme form exhibited an alkaline pH optimum, in contrast to the neutral pH optima of both soluble and weakly associated phosphorylase kinase. PMID- 8717431 TI - Effects of angiotensin II and aldosterone on collagen gene expression and protein turnover in cardiac fibroblasts. AB - Earlier studies have demonstrated angiotensin II (AngII) and aldosterone (ALDO) each augment cultured adult rat cardiac fibroblast (CFb) collagen synthesis. Whether this involves type I collagen, the major structural protein of the myocardium, and represents a transcriptional event, is uncertain. Accordingly, the influence of AngII and ALDO on transcription and synthesis of fibrillar collagen and on collagenolytic activity was examined in cultured CFb maintained in serum-deprived media. Using concentrations for AngII (10(-7) M) or ALDO (10( 9) M), shown to influence collagen turnover in these cells, we found: a) total collagen synthesis was significantly (p < 0.05) increased (5.4 +/- 0.41 and 4.8 +/- 0.37 vs. control 3.1 +/- 0.55); b) type I collagen production (6590 +/- 710 and 6150 +/- 410 vs. control 4700 +/- 490 ng/mL) in the medium were significantly (p < 0.01) increased; c) type I collagen mRNA expression was also significantly (p < 0.01) increased by AngII (2.0 fold) and ALDO (1.8 fold) compared with control; d) AngII, but not ALDO, significantly (p < 0.05) decreased collagenolytic activity (0.5 fold) compared with control. Thus, AngII and ALDO each increase CFb type I collagen synthesis at the level of transcription and protein synthesis and AngII, but not ALDO, alters collagenolytic activity. Such hormonally mediated alterations in CFb collagen turnover may contribute to the adverse accumulation of fibrillar collagen found in the myocardium in various disease states, where circulating AngII and/or ALDO are increased. PMID- 8717432 TI - Identification of guanine nucleotide binding regulatory proteins in bovine tracheal smooth muscle. AB - The identity of G-proteins in airway smooth muscle is not well elucidated. In the present study, by immunoblotting using AS/7 antibody specific for Gi alpha-1/2, EC/2 antibody specific for Gi alpha-3 and RM/1 antibody specific for Gs alpha, we identified, respectively, Mr 39, 41, 46 and 52 KDa, Mr 41 and 43 KDa, and Mr 43 and 46 KDa polypeptides of conventional (heterotrimeric) G-proteins in purified membranes of bovine tracheal smooth muscle. The identity of the Mr 41, 43 and 52 KDa Gi alpha, and the Mr 43 and 46 Gs alpha was also confirmed by ADP ribosylation with pertussis and cholera toxins, respectively. In addition, the common antibody (AG/1) for both Gi alpha and Gs alpha revealed the presence of all the above polypeptides, except the Mr 52 KDa band. By nitrocellulose blot overlay with [35S]s alpha GTP gamma S, we also detected seven low molecular weight GTP-binding proteins of Mr 18-30 KDa in the bovine tracheal smooth muscle. Photoaffinity crosslinking of [alpha-32P]GTP demonstrated the presence of high molecular GTP-binding proteins of Mr 55, 75 and 110 KDa. It is concluded that plasma membranes of bovine tracheal smooth muscle contain various types of conventional, low molecular weight and high molecular weight G-proteins. This warrantes further attention to elucidate the functional roles of G-proteins in airway smooth muscle. PMID- 8717433 TI - Role of cardiac renin-angiotensin system in the development of pressure-overload left ventricular hypertrophy in rats with abdominal aortic constriction. AB - Possible involvement of cardiac renin-angiotensin system (RAS) in pressure overload induced left ventricular hypertrophy (LVH) was investigated. Rats were subjected to abdominal aortic constriction (AAC) and examined the effects of 4 weeks treatments with an angiotensin converting enzyme (ACE) inhibitor, captopril and a vasodilator, hydralazine on haemodynamics and ventricular RNA, DNA, protein and myosin isoform pattern in sham and hypertrophied rats. AAC increased the mean arterial pressure (MAP) and systolic blood pressure (SBP), and resulted in increased left ventricle/body weight ratio, LV thickness, RNA and protein content, however total DNA was not changed. The expression of fetal isogene, beta myosin heavy chain (beta-MHC), was markedly enhanced where as alpha-MHC was reduced. High-dose captopril (100 mg/kg p.o.,) significantly prevented the increase in haemodynamics, development of LVH, LV remodeling, increase in total protein, RNA and antithetical expression of myosin isoforms. Hydralazine (15 mg/kg p.o.,), did not modulate hypertrophic changes and low-dose captopril (1.5 mg/kg p.o.,) which has not produced any marked fall in MAP and SBP also modulated favourably the development of LVH and its biochemical markers. Thus, the prevention of the development of LVH and induction of beta-MHC by non-hypotensive doses of captopril may be related to the blockade of intracardiac production of angiotensin II rather than circulating system. These results suggest that cardiac RAS may play an important role in pressure overload induced LVH. PMID- 8717434 TI - Matrix metalloproteinase activity expression in infarcted, noninfarcted and dilated cardiomyopathic human hearts. AB - In the normal myocardium matrix metalloproteinases (MMP) are present in the latent form. To examine whether MMP are activated following infarction or idiopathic dilated cardiomyopathy (DCM), we extracted and measured MMP activity in tissue derived from 7 explanted, failing human hearts due to either previous myocardial infarction (MI) or DCM. MMP activity in infarcted left ventricle (LV), noninfarcted LV and right ventricle (RV) from MI patients, as well as tissue from either ventricle of DCM patients, were compared to the activity of donor heart tissue. SDS-PAGE and dye-binding assays were used to determine total protein concentration, while collagenase activity was measured by SDS-PAGE type substrate gels embedded with type I gelatin (zymography). Accuracy of the zymographic technique was shown for tissue samples as small as 0.05 mg and was comparable to results obtained by a spectrophotometric method. After normalization for total protein concentration, we found 3 +/- 1% collagenase activity in normal atrial tissue which could be activated to 80-90% by trypsin or plasmin, indicating that collagenase is normally inactive or in a latent form in human heart. In endo- and epimyocardium of infarcted LV, on the other hand, collagenase activity was 85-95% and 10-20%, respectively, while 5-10% and 3-5%, respectively, in noninfarcted LV. In DCM, collagenolytic activity in the endo and epimyocardium was 75 +/- 5 and 35 +/- 5% in the LV and 35 +/- 7 and 20 +/- 5% in the RV, respectively. Thus, in dilated failing human hearts secondary to previous MI or DCM, MMP activity is increased. This is particularly the case within the endomyocardium of the infarcted and noninfarcted portions of either ventricle with MI and in both ventricles in DCM. This suggests that an activation of collagenase throughout the myocardium may contribute to its remodeling that includes ventricular dilatation and wall thinning. PMID- 8717435 TI - Purification and characterization of the carboxyl-domain of human hexokinase type III expressed as fusion protein. AB - In mammalian tissues hexokinase (ATP:D-hexose 6-phosphotransferase, EC 2.7.1.1) exists as four isoenzymes encoded by distinct genes. These proteins are homologous and are organized in two homologous domains, with the exception of hexokinase type IV which has only one. This organization is believed to be the result of a duplication and tandem fusion event involving the gene encoding for the ancestral hexokinase. In this study, we cloned the carboxyl-domain of human hexokinase type III and expressed it in Escherichia coli as a glutathione S transferase fusion protein, using the pGEX-2T expression vector. The recombinant protein showed catalytic activity. A comparative study of the kinetic properties of the expressed carboxyl-domain and the enzyme partially purified from human lymphocytes is also shown. The results now allow a better understanding of the role of the carboxyl-domain in determining the catalytic properties of the enzyme. PMID- 8717436 TI - Expression of c-Fos in subcutaneous adipose tissue of the fetal pig. AB - Subcutaneous adipose tissue from fetal pigs was examined for c-Fos expression in developing adipocytes. Enhanced c-Fos expression was found in the nuclei of adipocytes and cells closely associated with fat cell clusters from 75 and 105 day old fetuses. 50 day old fetuses which had no adipocytes showed no enhanced c Fos expression in undifferentiated cells. c-Fos expression in adipocytes was not transient, but persisted through the gestational period from 75 to 105 days. The timing of c-Fos induction appears to be closely related to adipocyte differentiation and was found only in developing adipocytes and cells closely associated with fat cell cluster and was not found in independent stromal vascular cells. PMID- 8717437 TI - Differential time course of liver and kidney glucose-6 phosphatase activity during long-term fasting in rat correlates with differential time course of messenger RNA level. AB - We have studied the role of Glc6Pase mRNA abundance in the time course of Glc6Pase activity in liver and kidney during long-term fasting in rat. Refered to the mRNA level in the fed state, Glc6Pase mRNA abundance was increased by 3.5 +/- 0.5 and 3.7 +/- 0.5 times (mean +/- S.E.M., n = 5) in the 24 h and 48 h-fasted liver, respectively. Then, the liver Glc6Pase mRNA was decreased to the level of the fed liver after 72 and 96 h of fasting (1.0 +/- 0.3 and 1.4 +/- 0.3). In the kidney, Glc6Pase mRNA abundance was increased by 2.7 +/- 1.0 and 5 +/- 1.2 times at 24 and 48 h of fasting, respectively. Then, it plateaued at the level of the 48 h fasted kidney after 72 h and 96 h of fasting (4.5 +/- 1.0 and 4.3 +/- 1.0). After 24 and 48 h-refeeding, the abundance of Glc6Pase mRNA in 48 h-fasted rats was decreased to the level found in the liver and kidney of fed rats. The time course of the activity of Glc6Pase catalytic subunit during fasting and refeeding was strikingly parallel to the time course of Glc6Pase mRNA level in respective tissues. These data strongly suggest that the differential expression of Glc6Pase activity in liver and kidney in the course of fasting may be accounted for by the respective time course of mRNA abundance in both organs. PMID- 8717438 TI - Gender-related regional antioxidant profiles in the gastrointestinal tract of the rat. AB - This study compared the activities of the antioxidant enzymes glutathione peroxidase, glutathione reductase and superoxide dismutase and the levels of glutathione in the mucosa of the body of the stomach, proximal and distal parts of the small intestine and the colon in male and female Sprague Dawley rats. Basal glutathione levels were significantly (p < 0.05) higher in the small intestine as compared with those in the other portions studied in both sexes. Except for colonic mucosa in females, the activity of glutathione reductase was similar in all the other tissues examined. Glutathione peroxidase showed the largest regional differences, with activities in the gastric segment being several-fold greater than those in small intestine or colon. This enzyme also showed marked gender-related differences, activity being greater in females than males in gastric mucosa and colon, while the converse was true for distal small intestine. In contrast, activities of superoxide dismutase showed minimal regional or gender-dependent variations. PMID- 8717439 TI - Thermal response in murine L929 cells lacking alpha B-crystallin expression and alpha B-crystallin expressing L929 transfectants. AB - We investigated the role of alpha B-crystallin expression in the development of thermotolerance in murine L929 cells. An initial heat-shock of 10 min at 45 degrees C induced thermotolerance in these cells to a heat challenge at 45 degrees C administered 24 h later. The thermotolerance ratio at 10(-1) isosurvival was 1.7. Expression of alpha B-crystallin gene was not detected during the 24 h incubation at 37 degrees C following heat shock by either northern or western blots. In contrast, inducible HSP70 synthesis was observed during this time period. Thus, this cell line provided an unique system in which to examine the effects of transfected alpha B-crystallin on thermoresistance and thermotolerance. Cells stably transfected with alpha B-crystallin under the control of an inducible promoter did not show a significant increase in the ability to develop thermotolerance. However, a stably transfected L929 clone expressing high levels of constitutive alpha B-crystallin exhibited an approximately 50% increase in thermal resistance over parental and control cells. Though expression of alpha B-crystallin is not requisite for the development of thermotolerance in L929 cells, overexpression of transfected alpha B-crystallin can contribute to increased thermoresistance. PMID- 8717440 TI - Effect of interleukin-1 receptor antagonist (IL-1RA) on histamine and serotonin release by rat basophilic leukemia cells (RBL-2H3) and peritoneal mast cells. AB - Recently, it has been appreciated that cultured mast cells are significant sources of cytokines. However, the role of interkeukin-1 (IL-1) on mast cells and/or basophil degranulation is still unclear. In this report we provide evidence that rat basophilic leukemia cells (RBLC) cultured with a natural inhibitor of IL-1, interleukin-1 receptor antagonist (IL-1RA) (500 ng/ml) for 48 h, strongly inhibited the spontaneous release of serotonin (5HT) and histamine (from 22.50 to 43.49%), compared to untreated cells (control). When IL-1RA treated and untreated RBLC were stimulated with a secretagogue (anti-IgE), no difference was found in the percent of 5HT and histamine release. Moreover, in another set of experiments using rat peritoneal mast cells (RPMC) treated and untreated with IL-1RA, we found that IL-1RA did not affect the release of 5HT or histamine, even when the secretagogue anti-IgE or compound 48/80 (C48/80) were used. The present studies describe an additional biological activity of IL-1RA, inhibiting histamine and 5HT release from RBLC cultures. PMID- 8717441 TI - Expression of human N-myristoyltransferase in Escherichia coli. Comparison with N myristoyltransferases expressed in different tissues. AB - Myristoyl CoA:protein N-myristoyltransferase catalyzes the addition of myristate to the amino-terminal glycine residue of a number of eukaryotic proteins. Escherichia coli transformed with human NMT expression construct produced high levels of N-myristoyltransferase. Using the combination of ammonium sulfate precipitation, chromatography on SP-Sepharose fast flow and fast protein liquid chromatography on Mono-S, the enzyme was purified more than 100 fold with 40% yield. The hNMT fusion protein exhibited an apparent molecular weight of 53 kDa on SDS-polyacrylamide gel electrophoresis. Upon cleavage by the Enterokinase [(Asp)4-Lys], the hNMT exhibited an apparent molecular mass of 49 kDa without loss of catalytic activity. The hNMT activity could be greatly activated severalfold with the use of Tris, SDS, ethanol and acetonitrile. The catalytic activity of hNMT was potently inhibited in a concentration dependent manner by NIP71, a bovine brain NMT inhibitory protein with a half maximal inhibition of 31.0 nM. The E. coli expressed hNMT was homogeneous and showed enzyme activity. PMID- 8717442 TI - Preferential uptake of long chain polyunsaturated fatty acids by isolated human placental membranes. AB - Fatty acid uptake by the placenta is thought to be a carrier-mediated process, however the mechanism by which long chain polyunsaturated fatty acids (LCPUFA) are preferentially accumulated from the maternal circulation to the fetal tissues is still unclear. To examine the role of the placenta in this process, binding of four different radiolabelled fatty acids (-14C-oleate, -14C-linoleate, [14C]a linolenate and [14C]arachidonate) to human placental membranes was studied. Binding of fatty acid was found to be time- and temperature dependent. At equilibrium, the total binding of oleate was highest (5.1 +/- 0.1 nmoles/mg protein) followed by linoleate (2.8 +/- 0.31 nmoles/mg protein) and arachidonate (2.06 +/- 0.4 nmoles/mg protein) and alpha-linolenate binding was lowest (0.5 +/- 0.1 nmoles/mg protein). However, oleate had the lowest specific binding (37% of the total binding) whereas arachidonate had the highest specific binding (approximately 86% of the total binding) followed by linoleate and a-linolenate (62%, and 69% of the total binding, respectively). Binding of each [14C] fatty acid was also assessed in the presence of 20-fold excess of other unlabelled ligands. Binding sites seem to have preference for the binding of [14C] fatty acids in the following order: arachidonic acid >>> linoleic acid >> a-linolenic acid >>>>> oleic acid, whereas BSP and a-tocopherol did not show any competition with any of the [14C] fatty acids. These data suggest that the fatty acid binding sites in placental membranes are specific for the fatty acids but that they have heterogeneous affinities. Trans fatty acids (elaidic and linoelaidic acids) also competed very strongly for the [14C] fatty acid binding. Polyclonal antiserum raised against placental FABPpm inhibited binding of these [14C] fatty acids but with variable degrees of inhibition; EFA/LCPUFA binding was much more than that of oleate. Our data suggest that EFA/LCPUFA bound to albumin are preferentially transported by human placental membranes and that the placental FABPpm may be involved in the sequestration of EFA/LCPUFA by the placenta. PMID- 8717444 TI - Regulation of endothelial IGFBP-3 synthesis and secretion by IGF-I and TGF-beta. AB - We have examined the regulation of endothelial IGFBP-3 production by IGF-I and TGF-beta, two growth factors thought to play a major roles in the complications of diabetes mellitus. In addition, we developed a sensitive method for IGFBP-3 mRNA quantitation by adapting the fluorescent modification of the competitive PCR strategy. Our results using both Northern analysis and the fluorescent competitive PCR method indicate that: (1) IGFBP-3 mRNA is increased 2- to 10-fold by IGF-I and maximally reduced to 20% of control by TGF-beta; (2) the changes in mRNA levels correlate with the levels of IGFBP-3 protein secreted into the media by these cells; (3) the induction of IGFBP-3 mRNA and protein by IGF-I analogs was directly related to their ability to bind to the type I IGF receptor, reflecting an IGF-I receptor-mediated process; and (4) steady state IGFBP-3 mRNA levels did not change significantly after a 6 h incubation with actinomycin D in the presence or absence of the growth factors suggesting that the observed IGF I/TGF-beta effects occur at the level of gene transcription rather than mRNA stability. PMID- 8717445 TI - Regulation of insulin-like growth factors and IGF-binding proteins in bone tumours. AB - Insulin-like growth factors (IGFs) and IGF-binding proteins (IGFBPs) are important regulators of growth and development in normal bone and bone tumours. IGFs contribute to about 50% of basal bone cell proliferation. Their mitogenic actions is either enhanced or inhibited by specific IGFBPs which are also regulated by a variety of factors. This system is further complicated by the presence of specific proteases for IGFBPs. Serine proteases are secreted by malignant bone tumours and the hydrolysis of IGFBPs increases the bioavailability of IGFs. In addition, the autocrine production of IGFs may facilitate the development of bone tumours and metastatic lesions. PMID- 8717443 TI - Expression of calcium-binding protein regucalcin mRNA in hepatoma cells. AB - Whether the gene expression of hepatic Ca(2+)-binding protein regucalcin is altered in hepatomas was investigated. The change in regucalcin mRNA levels was analyzed by Northern blotting using liver regucalcin complementary DNA (0.9 kb). Rat hepatoma was induced by continuous feeding of basal diet containing 0.06% 3' methyl-4-dimethylaminoazobenzene (3'-Me-DAB). After 35 weeks feeding, rats were sacrificed, and the non-tumorous and tumorous tissues of the livers were removed. In individual rats, the regucalcin mRNA levels in the tumorous tissues were generally decreased in comparison with that of the non-tumorous tissues of the chemical-fed rats, although the chemical administration might decrease the mRNA expression in normal rat liver, suggesting that the chemical administration causes a suppressive effect on the mRNA expression. When the genomic DNA extracted from the liver tumorous tissues was digested with restriction enzymes (EcoRI, BamHI and HindIII) and analyzed by Southern blotting, no rear-ranged band was found in the regucalcin gene from the hepatoma. Interestingly, in the transplantable Morris hepatoma cells, the regucalcin mRNA was markedly expressed, while the albumin mRNA was expressed only slightly. The present study demonstrates that regucalcin mRNA is clearly expressed in the transformed cells (Morris hepatoma cells). PMID- 8717446 TI - Ontogeny of growth hormone protein and mRNA in the gilthead sea bream Sparus aurata. AB - The ontogeny of growth hormone (GH) protein and mRNA was studied in the gilthead sea bream Sparus aurata by Northern blot, reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analyses. Total RNA was prepared from a pool of larvae collected on different days after hatching. When hybridized to Sparus aurata GH cDNA, GH specific mRNA was first seen on day 3 post-hatching. Levels increased on subsequent days and reached the highest levels on day 9; thereafter a decrease was noted to similar levels between days 10 and 16. RT-PCR amplification of larval RNA showed an amplified fragment on day 3, which hybridized to Sparus aurata GH cDNA but not on days 1 and 2 after hatching. Soluble proteins were electrophoresed and immunoreacted with antibodies raised against purified Sparus GH. A faint band was detected on day 2 after hatching, which had an electrophoretic mobility very similar to recombinant tilapia GH or Sparus pituitary extract. The immunoreactive material increased on days 3, 4 and 6. Specificity of the immunoreaction was demonstrated by the ability of recombinant tilapia GH to complete with GH bound to the membrane and inability of normal rabbit serum (NRS) to detect GH on the membrane. Our results suggest that both GH mRNA and protein are expressed in Sparus aurata shortly after hatching. PMID- 8717447 TI - KGF and EGF differentially regulate the phenotype of prostatic epithelial cells. AB - Previous studies indicate that keratinocyte growth factor (KGF) acts as a paracrine factor in the prostatic epithelium and epidermal growth factor (EGF) acts as an autocrine factor. In serum-free medium, KGF or EGF promoted similar growth of human prostatic epithelial cells. Response to two growth-inhibitory factors (suramin and transforming growth factor-beta), and expression of keratins and prostate-specific antigen (PSA), were similar with either mitogen. However, colonies in medium with KGF were very compact with extensive intercellular bonds, whereas colonies with EGF consisted of widely-separated cells. Growth was decreased to a greater extent by deletion of growth factors from medium with KGF versus EGF, and retinoic acid was 10-fold more potent at inducing growth inhibition and differentiation-associated keratin with KGF compared with EGF. We conclude that regulation of growth and differentiation in the prostate might vary depending on the availability of KGF versus EGF. PMID- 8717448 TI - Rapid clearance of human insulin-like growth factor binding protein-3 from the rat circulation and cellular localization in liver, kidney and stomach. AB - The gastrointestinal tract represents a major site for the trophic actions of insulin-like growth factors (IGFs), which may be derived in vivo from a large circulating pool. The high capacity binding protein for IGFs in blood is IGF binding protein-3 (IGFBP-3), which is largely complexed with an acid-labile subunit (ALS). However, we and others have shown that IGFBP-3 not complexed with ALS can rapidly leave the circulation, and may carry IGFs to peripheral tissues. In this study we investigated the transfer of recombinant, glycosylated human (h)IGFBP-3 from the rat circulation to the stomach and intestine, compared with liver and kidney. [125I]-labeled IGFBP-3 was administered into the tail vein of conscious male rats, which were killed between 5 min and 2 h later. Blood was taken for the preparation of plasma, and the liver, kidneys, stomach and intestine were removed either for estimation of the associated radioactivity, or fixed for autoradiographic analysis of histological sections. Following injection, [125I]-labeled IGFBP-3 was associated, in part, with a 150 kDa complex in plasma within 10 min when analyzed by gel filtration chromatography. However, 84% of the administered IGFBP-3 had already left the circulation, and 40% of the initial injected dose was accumulated in liver by 5 min, with a further 4% localized in the kidneys. Autoradiographic analysis showed that IGFBP-3 was selectively accumulated within Kupffer cells of the liver, and by the glomeruli and proximal tubules of the kidney. Little radiolabeled IGFBP-3 was recovered from the small intestine, but 14% of the initial injected dose was found within the stomach after 2 h, and a further 12% within the stomach contents. Autoradiographic localization within the stomach showed that the [125I]-labeled IGFBP-3 was primarily associated with the mucosal lining and gastric glands. Separation on sodium dodecyl sulphate polyacrylamide gel electrophoresis showed that the majority of the radioactivity associated with the stomach contents represented small, degraded peptides. These results suggest that while a rapid clearance of IGFBP-3 is achieved by the liver and kidney, a longer term accumulation occurs in the stomach with a luminal secretion. This may represent a delivery system by which circulating IGFs may reach gastric tissue. PMID- 8717449 TI - Insulin-like growth factor binding protein-3: factors affecting binary and ternary complex formation. AB - IGF binding protein-3 (IGFBP-3), the major serum carrier of IGF-I and IGF-II in adults, circulates predominantly in a ternary complex with IGF and the acid labile subunit (ALS). The affinities of IGF and ALS binding have not previously been determined under conditions of temperature, ionic strength and pH which approximate those in the circulation. IGF-I and -II binding was optimal at pH 4.0 5.5, and relatively poor at pH 7.4. The addition of 0.1 mol/l NaCl to 50 mmol/l phosphate buffer increased the affinity of IGF-II for IGFBP-3 by 2-fold, and of IGF-I by 4-fold. ALS binding peaked at pH 5.5-6.0, and was markedly reduced at pH 7.4, where only approximately 0.4 mol ALS was bound per mol IGFBP-3, with greatly reduced affinity. ALS affinity was further reduced at 37 degrees C compared to 22 degrees C, and in the presence of 0.1 mol/l NaCl compared to its absence. Under 'physiological' conditions, the affinities of ALS binding to IGF-I-IGFBP-3 and IGF-II-IGFBP-3 complexes (2.5 x 10(8) l/mol and 5.8 x 10(7) l/mol, respectively) are 300-fold and 2000-fold lower than the constants for the formation of the corresponding binary complexes. Thus, ALS binding to IGFBP-3 complexes is much weaker than previously recognised, emphasising the importance of ALS dissociation as a controlling factor in the regulation of IGF bioavailability. PMID- 8717450 TI - The effect of epidermal growth factor on circulating levels of free and total IGF I and IGF-binding proteins in adult rats. AB - In neonatal rats, systemic administration of epidermal growth factor (EGF) results in reduced body weight gain and decreased levels of circulating IGF-I, which suggests that it be involved in the EGF-induced growth retardation. We investigated the effect of 4 weeks of EGF administration on circulating free and total IGF-I and IGF-binding proteins (IGFBPs) in adult rats treated with saline (controls), 30 (low dose group) and 150 (high dose group) microgram/kg/day EGF. Serum IGF-I was determined in ultrafiltrates (free) and acid-ethanol extracts (total), and serum IGFBPs using Western ligand blotting, which yielded four distinct molecular bands. The IGFBPs were tentatively identified as IGFBP-3 (a double band at 42 and 38 kDa), IGFBP-1 and/or IGFBP-2 (a single band at 30 kDa) and IGFBP-4 (a single band at 24 kDa). EGF administration did not change the body weight, tibia length, or liver, heart and lung weight. In contrast, serum total IGF-I and IGFBP-3 decreased dose-dependently: total IGF-I averaged 1470 +/- 100 micrograms/l (controls; mean +/- SEM), 1030 +/- 60 micrograms/l (low dose group; P < 0.005) and 760 +/- 40 micrograms/l (high dose group; P < 0.005), whereas differences between IGFBP-3 levels reached significance (P < 0.05) between controls and high dose rats, only. When compared to controls, levels of IGFBP-1 and/or IGFBP-2 were increased in the low dose group (P < 0.05), but unchanged in the high dose group. IGFBP-4 was unaffected by EGF. Free IGF-I averaged 74 +/- 6 micrograms/l in controls, and was reduced to 35 +/- 6 micrograms/l (low dose group; P < 0.005) and 57 +/- 5 micrograms/l (high dose group; P < 0.05). Free IGF I was inversely correlated (r = -0.49, P < 0.05) with IGFBP-1 and/or IGFBP-2. We conclude that in adult rats prolonged EGF administration has a marked depressing effect on circulating total and free IGF-I. Nevertheless, we did not observe any somatic growth retardation. PMID- 8717451 TI - Prostate Cancer: Current Issues, New Approaches. Proceedings of an international congress. London, England, October 19-21, 1995. PMID- 8717452 TI - Incidence of prostate cancer will double by the year 2030: the argument for. AB - Prostate cancer is a frequent cancer in old men which increases with age and is associated with wide geographical variation. Even if age-specific incidence rates remain stable, the problem of prostate cancer seems certain to increase in absolute terms simply because of the ageing of the population. This will have major economic implications for the future, particularly in Westernised societies, with any temporal trend in risk serving only to further compound the problem. However, since the risk of dying from prostate cancer has remained unchanged in many countries throughout this century, it may be that the increased incidence may be the result of increasing transurethral resection of the prostate procedures or screening techniques which have increased the detection and reporting of prostate cancer. PMID- 8717453 TI - Incidence of prostate cancer will double by the year 2030: the argument against. AB - No one doubts that the incidence of histologically diagnosed prostate cancer will increase markedly in the future; the increasing ageing population, greater awareness, and meticulous pathological search will all confirm and exaggerate known trends. The crucial question of whether the incidence of biologically significant prostate cancer diagnosed in asymptomatic younger men (< 65 years) will double by 2030 and whether urologists are comfortable with the concept of radical therapy for each and every one of these anxious patients remains debatable and unanswered at this time. PMID- 8717454 TI - Healthcare resources should be targeted to chemoprevention in prostate cancer: the argument for. AB - The search for localised disease increases the incidence of histologically determined prostate cancer, termed 'minimal' disease. Moreover, the number of locally advanced cases found at surgery increases the push towards surgery at earlier stages of disease. We already have knowledge of risk factors and epidemiological autopsy incidences, and have developed hypotheses to prevent clinical prostate cancer, therefore we should now be performing chemoprevention studies in time to save the next generation from unneeded treatment and healthcare deficits. PMID- 8717455 TI - Healthcare resources should be targeted to chemoprevention: the argument against. AB - Chemoprevention of prostate cancer is an attractive idea that appeals to many struggling with the treatment of this disease. Many details inherent in any preventive strategy must be considered before embarking on this course, no matter how seductive it may appear. Critical issues of effectiveness, cost, difficulties in implementation, side effects, duration of the preventive cycle, and public acceptance may thwart prevention of prostate cancer from being implemented in the near future. PMID- 8717456 TI - Screening and early detection of prostate cancer will decrease morbidity and mortality from prostate cancer: the argument for. AB - Since prostate cancer represents the most common male malignancy and the second most frequent cause of cancer-related mortality, there is a significant need for methods to either reduce the incidence, improve therapy, or provide early detection. Of these, only the latter appears to be currently at hand. Many investigators have demonstrated that early detection programmes, incorporating a carefully performed digital rectal examination and prostate-specific antigen level testing, have resulted in a significant detection rate of clinically relevant malignancy at a time when it is curable. Although problems such as overdetection, lead- and length-time bias, and significant financial burden are associated with prostate cancer screening, if we are to significantly impact the mortality from this disease, such approaches are probably warranted. PMID- 8717457 TI - Screening and early detection of prostate cancer will decrease morbidity and mortality from prostate cancer: the argument against. AB - The ability of prostate specific antigen, digital rectal examination and transurethral ultrasound, either individually or in combination, to detect unsuspected prostate cancer is beyond doubt. Early detection programmes have revealed prostate cancer in 6% of men over 50 years of age. Most are stage T1c, or confined to the prostate but, protagonists claim that, on the basis of size, they are 'clinically significant'. Mortality from this disease should be reduced by their treatment. Available statistics suggest that the lifetime incidence of 'screen-detected' cancers will far exceed the likelihood of dying from prostate cancer. Given the current operative mortality of total prostatectomy and the physical and psychological morbidity of screening, biopsy and treatment, it is far from certain that mortality or morbidity from prostate cancer will be decreased. Long-term outcome data for T1c prostate cancer do not exist. Is the likelihood of prostate cancer death the same for prostate-specific antigen thresholds of 2, 3 or 4 ng/ml, or from tumours found in 1 of 6, or 1 of 12 needle biopsies? None of this is known. European men deserve the benefit of scientifically based information before being exposed to another North American fashion. Prospective randomised trials investigating mortality, quality of life and cost benefit are the only solution. PMID- 8717458 TI - The risks outweigh the benefits of radical prostatectomy in localised prostate cancer: the argument for. AB - Prostate cancer represents a major menace to Western society. Since it is asymptomatic in its early stages and there is no curative therapy for advanced stages, our only hope to decrease the mortality rate from prostate cancer is through annual prostatic evaluations of men who agree to undergo such testing. It is important to emphasise that these patients should be carefully informed about the possible consequences. Since prostate cancer is a slow growing tumour and its clinical significance decreases with advancing in age, only young men with a life expectancy of 10-15 years or more should participate in early detection efforts. These men should be evaluated as diligently as possible through the combined use of digital rectal examination and the newly described prostate-specific antigen testing, i.e. analysis of the ratio between different forms of serum prostate specific antigen. Radical prostatectomy, when performed on men with a life expectancy of 15 years or more who truly have organ-confined prostate cancer, is an acceptable and effective treatment for this group of patients who would benefit most from a radical approach to their disease. PMID- 8717459 TI - The risks outweigh the benefits of radical prostatectomy in localised prostate cancer: the argument against. AB - The value of early detection, via screening and/or treatment of localised prostate cancer has not been established in well-designed clinical trials. However, radical prostatectomy in localised disease can be justified for a number of reasons. Cure of localised prostate cancer is necessary: 3% of men over 50 years of age will die of prostate cancer. Cure is possible: prostate tumours take 10-15 years to grow from 0.5 cm3 to, for example, 8 cm3--the window where cure is possible by radical prostatectomy. Large series of studies show that the probability of remaining progression free, as determined by prostate specific antigen, can be up to 90% for pT2 tumours and 60-75% for pT3a/b tumours. A radical, anatomic technique is necessary with a low positive margin rate (zero in pT2 tumours) to achieve such results. Radical prostatectomy has a low morbidity: perioperative mortality is far below 1%, complete continence rate is 80-90%, complete incontinence rate 4%, stricture rate 6%, and relaparotomy rate <5%. To conclude, in suitable patients (for example, younger than 70 years of age) with tumours at suitable stages, cure is necessary, possible, and safe using the right technique of radical prostatectomy. PMID- 8717460 TI - Combined androgen blockade is the treatment of choice for patients with advanced prostate cancer: the argument for. AB - Combined androgen blockade (CAB) is becoming standard treatment for patients with newly diagnosed advanced prostate cancer. This statement is based on the outcome and long-term evaluation of patients treated in prospective randomised trials. Particularly, patients with 'good' prognostic signs at the time of diagnosis benefit most from this approach. Patients with metastatic prostate cancer treated with CAB can be stratified on the basis of clinical prognostic parameters such as performance status, the presence or absence of pain, the levels of alkaline phosphatase, and the levels of prostate-specific antigen (PSA) during the first months of CAB: for combined androgen deprivation, if they fall in the good prognostic category (+/- 30-35% of the patients); for surgical castration and palliative treatment or (experimental) cytotoxic therapy if they belong to the poor prognostic category (+/- 20% of the patients), and, for initial CAB during the first 3-6 months of therapy, followed by an evaluation of the regression of PSA under therapy. If PSA normalises (rapidly) during this period, patients could be considered as good prognostic candidates and continue CAB. If PSA does not decrease or normalise sufficiently, patients might be classified as belonging to the poor prognostic category and should be treated with palliative measures and/or experimental cytotoxic therapy after (surgical) castration. PMID- 8717461 TI - Combined androgen blockade is the treatment of choice for patients with advanced prostate cancer: the argument against. AB - The concept that patients with advanced prostate cancer will survive longer if treated by a combination of castration and the blockade of androgens of non testicular origin cannot be substantiated. When all of the available studies are reviewed, meta-analysis does not show any significant improvement in mortality when the combined approach is used. Some studies have suggested that combined treatment is particularly useful in patients with minimal disease, or with a good prognosis; it must be remembered that these are smaller groups which have been taken from large numbers of patients in which the endpoint was observations on the larger group and not the small subgroup. PMID- 8717462 TI - Supportive care is not the only option in prostate cancer patients resistant to hormone therapy: the argument for. AB - In multivariable analyses conducted by several groups, a post-therapy decline in prostate specific antigen was the most significant factor associated with survival of patients with relapsed prostate cancer. Contemporary combination regimens result in consistent declines in prostate-specific antigen in up to 40% of cases, along with objective tumour regression, and the improvement in cancer related symptoms. It is, therefore, apparent that prostate cancers are not as resistant to treatment as previously believed. PMID- 8717463 TI - Supportive care is not the only option in prostate cancer patients resistant to hormone therapy: the argument against. AB - Hormone-resistant prostate cancer patients are elderly, frail and in pain. They have a median survival of 6 months. There is no convincing evidence from controlled trials that anything we do will increase life expectancy. Any attempt to do so with currently available agents may either kill them earlier or decrease the quality of the short life left to them. The alternatives for management include the simple, non-toxic, supportive measures of better analgesic use, antiandrogen withdrawal, external beam radiation and steroids, which can produce significant symptomatic improvement. There is little evidence that the benefits of more aggressive therapy exceed those achieved with supportive care. PMID- 8717464 TI - Epidemiology, prevention and screening for prostate cancer. AB - Prostate cancer is a growing social problem. The economical impact is high and is likely to become greater in future years, not only because the number of patients diagnosed with prostatic neoplasias is increasing steadily, but also because of the promotion of screening programmes and the rise in the number of surgical procedures available. It can be concluded that, at the present stage, widespread implementation of screening cannot be recommended. The answer to the question of screening can only be provided by large randomised trials, comparing either screening versus no screening, or early versus delayed treatment in patients in whom early cancer is detected by screening. PMID- 8717465 TI - Combined androgen blockade. AB - Combined androgen blockade (CAB), the addition of an antiandrogen to castration (medical or surgical), is becoming a well-established option for advanced prostate cancer. CAB with leuprolide and flutamide was found to improve both progression-free and overall survival, compared with leuprolide alone, particularly in patients with minimal metastatic disease. These findings have led to an intergroup study in more than 1,300 men, including 300 with minimal disease, of orchidectomy with flutamide versus orchidectomy with placebo, which will examine which patient subgroups benefit most from CAB. This intergroup study has already provided important information regarding prostate-specific antigen as a surrogate marker for response and progression. The success with CAB in lengthening survival and improving quality of life in metastatic prostate cancer has stimulated enthusiasm for broadening the base of clinical studies for the management of this disease. PMID- 8717466 TI - The management of localised cancer of the prostate. AB - Localised prostate cancer (T1/T2) can be managed using a conservative (expectant) approach or by radical treatment with curative intent. Expectant management involves observation until symptoms of local/distant progression appear. Radical treatment with intent to cure is based on the destruction (radiation therapy or cryosurgery) or removal (surgery) of the tumour. Many options are offered to patients with T1/T2 prostate cancer. Expectant therapy may be considered in patients with limited life expectancy, curative treatment should be considered in the others, taking into account the high rate of cancer-related deaths after 10 years. Moreover, accurate staging should lead to a better selection of patients likely to be cured by monotherapy (surgery or radiation therapy depending on the patient's and doctor's biases). Surgery probably offers the advantage of the certainty of removal over the hypothetical destruction of the whole tumour provided by radiation therapy. PMID- 8717467 TI - The management of hormone refractory prostate cancer. AB - Prostate cancer patients deemed unsuitable for curative therapy by means of radical prostatectomy or radiation are offered androgen ablation or suppression therapy, which normally results in an inhibition of prostate cellular growth for a period of 12-18 months. Death of patients from prostate cancer is directly related to the development of clones of cells capable of multiplying and metastasising without androgen stimulation, and to date efforts to suppress these cells have been singularly unsuccessful. There are a number of treatment options available, and these include secondary hormone therapy, growth factor inhibition, chemotherapy, combination therapies and interstitial radiotherapy. The merits and disadvantages of these management approaches are discussed. PMID- 8717468 TI - Androgens, antiandrogens and androgen receptor abnormalities. AB - Information on the molecular structure of the human androgen receptor has increased insight into the molecular mechanism of action of androgens and antiandrogens. It has also facilitated the study of molecular defects in the androgen receptor gene associated with prostate cancer. Several somatic mutations have been detected in tumour specimens of patients with prostate cancer. Most of the reported mutations are localised in the ligand binding domain. A relatively high frequency of the Thr868Ala mutation (originally reported for the human prostate cancer cell line androgen receptor) is particularly found in metastatic lesions (bone metastases) of prostate cancer and can be considered as a hot spot. It could be speculated that this specific mutant androgen receptor provides a selective growth advantage in a subset of advanced prostate cancers. For a limited number of mutations it has been shown that ligand responsiveness to adrenal androgens, progestagens, oestrogens and some antiandrogens of the mutant receptors has been increased. The consequences of these mutations could be that the androgen receptor can still be activated in castrated patients and during antiandrogen therapy. The observation, therefore, that antiandrogen withdrawal can be beneficial for some prostate cancer patients, might be understood in the light of an altered ligand responsiveness of mutant androgen receptors. PMID- 8717469 TI - The development of Casodex (bicalutamide): preclinical studies. AB - Casodex (bicalutamide, Zeneca Limited) was developed for the treatment of prostate cancer from a series of nonsteroidal compounds related to flutamide. Casodex is a selective antiandrogen that binds to rat, dog and human prostate androgen receptors, and has approximately a 4-fold higher affinity for the rat androgen receptor than hydroxyflutamide, the active metabolite of flutamide. Casodex also binds to androgen receptors found in the LNCaP human prostate tumour and the Shionogi S115 mouse mammary tumour cell line, as well as androgen receptors transfected into CV-1 and HeLa cells. In all cases, Casodex behaves as a 'pure' antiandrogen and inhibits gene expression and cell growth stimulated by androgens. Studies in vivo show that Casodex is a potent antiandrogen in the rat. In contrast to flutamide, which produces dose-related, marked increases in serum luteinising hormone (LH) and testosterone, Casodex has little effect on serum LH and testosterone; that is, it is peripherally selective. The peripheral selectivity of Casodex has now been shown to be due to poor penetration across the blood-brain barrier. In dogs, Casodex has exquisite potency and causes dose related atrophy of the prostate gland and epididymis; with an oral ED50 of 0.1 mg/kg, it is about 50 times as potent as flutamide in this species. Casodex is also peripherally selective in the dog. In addition, magnetic resonance imaging studies have shown that Casodex is a potent antiandrogen in the monkey. Casodex, at a daily oral dose of 25 mg/kg effected a highly significant reduction in the growth of Dunning R3327H transplantable rat prostate tumours that was equivalent to that achieved by either surgical or medical castration with the LH-releasing hormone agonist Zoladex (goserelin). In a comparative study, flutamide was shown to be both less potent and less active than Casodex. In these preclinical studies, Casodex was well tolerated. The preclinical properties of Casodex give it advantages, with respect to potency, tolerability and the maintenance of effective antiandrogen serum concentrations, over other available antiandrogens. Moreover, it has a half-life that is compatible with once-daily administration. PMID- 8717470 TI - Clinical progress with a new antiandrogen, Casodex (bicalutamide). AB - Casodex (bicalutamide, Zeneca Limited) is a nonsteroidal competitive inhibitor of androgens at the androgen receptor. The drug was developed to fulfil a number of needs for the treatment of prostate cancer. The specific aim was to demonstrate pharmacological activity, which would translate into clinical efficacy, good tolerability in the context of its use, oral availability, a convenient and forgiving dosing regimen, and clinical acceptability. Casodex has been shown to be orally bioavailable and well absorbed, with a plasma half-life of around 1 week. Although steady-state levels are not reached for 1 month, there is evidence that the androgen receptor blockade achieved with Casodex is equivalent to that of flutamide by the end of the first day. The dose of Casodex was established in a series of dose-ranging studies using the surrogate endpoints of prostatic acid phosphatase (PAP) and prostate-specific antigen (PSA). Initial studies suggested that a dose of 50 mg daily gave a fall in PAP equivalent to that seen with castration. This dose was, therefore, evaluated as monotherapy and subsequently as a component of combined androgen blockade. Higher doses were evaluated using PSA as a surrogate endpoint and, although doses up to 450 mg have been given to man, 150 mg daily is well tolerated with demonstrable evidence of activity. Although trials of the drug at 150 mg in monotherapy have, to date, not shown survival equivalence with castration, Casodex has been well tolerated with evidence of good symptomatic response and quality-of-life benefits including the potential of retaining libido. In combination treatment, Casodex is associated with significantly less gastrointestinal effects (diarrhoea) than the nonsteroidal antiandrogen flutamide (Eulexin, Schering-Plough International). Casodex is not associated with alcohol intolerance, pneumonitis and ocular defects which have been seen with the antiandrogen nilutamide (Anandron, Roussel). Moreover, since Casodex is a nonsteroidal antiandrogen, no steroidal effects have been seen. PMID- 8717472 TI - Appraisal of current therapies for prostate cancer. AB - All aspects of the treatment of prostate cancer remain controversial. Even the simplest question--is early detection of prostate cancer beneficial?--remains the subject of heated debate. While it would appear intuitive that early detection should increase the survival of patients with prostate cancer, no high quality evidence actually supports this assumption. Indeed decision-analysis models have suggested that early detection and treatment may impair quality-adjusted survival. More practical issues such as the effectiveness and side effects of traditional treatments for prostate cancer, such as radical therapy for localised disease and androgen ablation therapy, are now under great scrutiny. There are proponents and opponents of all treatments brandishing widely divergent analyses designed to support their views. In an era of cost containment these various treatments are all being subjected to outcome analyses which will determine their true benefit. Nonetheless, the judicious selection of patients most likely to benefit from each individual therapy will assure the best possible outcome. PMID- 8717471 TI - A controlled trial of Casodex (bicalutamide) vs. flutamide, each in combination with luteinising hormone-releasing hormone analogue therapy in patients with advanced prostate cancer. Casodex Combination Study Group. AB - Between January 1992 and September 1993, 813 patients with stage D2 prostate cancer were enrolled in a multicentre, double-blind (for antiandrogen therapy) trial and randomised to antiandrogen therapy with Casodex (bicalutamide, 50 mg once daily) or flutamide (250 mg three times daily) and to luteinising hormone releasing hormone (LHRH) analogue therapy with Zoladex (goserelin, 3.6 mg every 28 days) or leuprolide (7.5 mg every 28 days). Time to treatment failure was the primary efficacy endpoint. At a median follow-up time of 49 weeks, there was a significant (p = 0.005) difference between groups in time to treatment failure in favour of Casodex plus LHRH analogue. Overall, 168 (42%) of 404 patients in the Casodex plus LHRH analogue group and 218 (53%) of 409 patients in the flutamide plus LHRH analogue group reached a treatment failure endpoint. Although a cause specific treatment-failure analysis was not performed, the difference between groups in treatment failure attributed to adverse events (mainly diarrhoea) was evident primarily in the first 7 months of therapy. The difference between groups in treatment failure for objective progression was most evident after 1 year of therapy. With further follow-up (median time of 95 weeks), the result for time to treatment failure, although no longer statistically significant, were consistent with the previous finding of an improvement in time to treatment failure associated with Casodex plus LHRH analogue therapy. With a median of 95 weeks of follow-up, 34% of deaths had occurred. The survival analysis was not dissimilar between the 2 groups. At 49 weeks median follow up, the incidence of diarrhoea was significantly (p < 0.001) lower among patients in the Casodex plus LHRH analogue group. Diarrhoea led to withdrawal from therapy for 2 patients in the Casodex plus LHRH analogue group, compared with 25 patients in the flutamide plus LHRH analogue group. In conclusion, Casodex plus LHRH analogue is well tolerated and effective with an improvement in time to treatment failure over flutamide plus LHRH analogue. Survival was not dissimilar between the 2 treatment groups. PMID- 8717473 TI - The role of induction androgen deprivation prior to radical prostatectomy. AB - In patients with localised prostate cancer and a life expectancy of at least 10 years, radical prostatectomy is increasingly being performed. Pre-operative clinical staging of prostate cancer is, however, inaccurate with around 10-20% of clinical T1, 10-30% of clinical T2a, and 30-60% of clinical T2b cases having tumour at the surgical margin. There is no documented best treatment for patients with a positive surgical margin. Options include close observation, adjuvant radiation therapy or androgen deprivation. The role of induction androgen deprivation has, therefore, evolved in an attempt to improve upon the cure rate for radical prostatectomy, and in this review results from clinical trials are discussed. In the largest United States randomised trial in patients with stage T2b prostate cancer, pre-operative androgen deprivation resulted in a significantly lower rate of positive surgical margins, normalisation of prostate specific antigen and a decrease in the size of the prostate. Sufficient follow-up is required from all of the completed prospective randomised trials before pre operative androgen deprivation can be justified in all T2b patients. PMID- 8717474 TI - Quality of life in prostate cancer: what are the issues and how are they measured? AB - Improvement of quality of life (QoL) has become a major endpoint in clinical trials of patients with prostate cancer. However, there still exist considerable methodological problems regarding the development of optimal instruments and methods for presenting the results. Within the European Organisation for the Research and Treatment of Cancer (EORTC) a core questionnaire is generally used for QoL evaluation, combined with a treatment- and disease-specific module. So far, no official EORTC prostate cancer module has been developed. Previous and ongoing trials in prostate cancer have addressed the following issues: for localised prostate cancer, micturition and sexuality, and for metastatic prostate cancer, bone pain, micturition, sexuality, vitality, hot flushes and gynaecomastia. The future challenge is to incorporate QoL results into the overall evaluation of treatment in combination with survival results and economical considerations. PMID- 8717475 TI - Patient compliance with crucial drug regimens: implications for prostate cancer. AB - Understanding of patient compliance with crucial drug regimens has improved markedly since 1986, based on data from two objective methods for monitoring drug dosing by ambulatory patients. Electronic monitoring records times and dates of drug package use, and chemical markers, incorporated into drug dosage forms, are assayed in plasma. These methods remove the camouflage that masks many poor compliers. In contrast, other methods (returned tablet counts, interviews, questionnaires) allow patients easily to censor evidence for delayed or omitted doses. The new methods show many more and larger errors of omission in both trials and practice than previously believed. One patient in about six is punctually compliant, but a modest majority of patients make errors probably too small to attenuate or otherwise modify the actions of all but the most unforgiving medicines. About a third of patients delay or omit many prescribed doses, thus attenuating or otherwise modifying the actions of all but the most forgiving drugs. One patient in about six takes little medicine, though camouflaged as a good complier. Similar patterns of delayed and omitted does prevail, essentially independent of drug, disease, prognosis, or symptoms. In summary, patients take the prescribed dose at intervals longer than prescribed- often by hours, sometimes by days, occasionally by weeks. The clinical and economic consequences of these lapses in dosing are unique to the treatment situation and the severity of disease and comorbidity. The new methods have not yet been applied to androgen-blocking agents, but if the findings resemble those with, e.g. tamoxifen in breast cancer, it will doubtless trigger some rethinking about failed treatment, trial design, and clinical management. PMID- 8717476 TI - The future management of prostate cancer? AB - In improving the management of prostate cancer, cryotherapy, interstitial laser ablation and high intensity focused ultrasound may prove to be preferable to radical prostatectomy. For patients with advanced, metastatic or recurrent disease, the mainstay of therapy is likely to remain combined androgen blockage, that is the use of a luteinising hormone-releasing hormone analogue and antiandrogen combination. Other possible new therapeutic advances include the use of growth factor inhibitors such as suramin in hormone escaped prostate cancer and the use of gene therapy. In conclusion, the best hopes for managing prostate cancer lie in earlier detection of the disease, and there is a need for improved diagnostic indicators. It is also important, as a general principle, to increase the efficacy and reduce the morbidity of therapies. However, increased survival is not the only issue as improvements in quality of life are fundamental in patients with prostate cancer. PMID- 8717477 TI - Anna Freud: her life and her biography. PMID- 8717478 TI - Predictors of outcome in child psychoanalysis: a retrospective study of 763 cases at the Anna Freud Centre. AB - Under Anna Freud's guidance, the Anna Freud Centre developed a rigorous approach to the collection of child psychoanalytic data. Material contained in detailed diagnostic assessments and weekly written reports of 763 cases treated in intensive and nonintensive therapy has now been subjected to systematic study. This is the first, retrospective stage of a major investigation of child psychoanalytic outcome, carried out in collaboration with Yale University Child Study Center, New Haven, CT. The main findings of the work are reviewed. The study showed child analysis to be particularly effective for seriously disturbed children under 12 years suffering from a variety of psychiatric disorders, particularly those which involve anxiety. PMID- 8717479 TI - Language, communication, and transference in child analysis. I. Selective mutism: the medium is the message. AB - Selective mutism is a psychiatric condition in which children stop speaking to all except their closest family members. Analytic material from a 4 1/2-year-old selectively mute child demonstrates how his symptom of mutism was embedded in a character structure in which not only verbal dialogue but play and other nonverbal communication were disrupted. Because communication was severely limited, I modified my analytic technique. Despite obstacles to communication, conflicts from different developmental levels were revived in the transference. Even in these difficult circumstances, analytic work in the transference resulted in significant change. PMID- 8717480 TI - Language, communication, and transference in child analysis. II. Is child analysis really analysis? AB - Is child analysis really analysis? The child patient does not free associate, communicates to the analyst in large measure nonverbally, and operates from a different psychic organization than the adult. Nevertheless, child and adult analysis share the same basic principles, despite different techniques. Using clinical examples from Jeremy's analysis, I discuss how child analysis could benefit from greater use of analysis of transference and analysis of defense. I then discuss recent questions about the place of verbalization and interpretation in child analysis. This issue parallels polarities in adult theory concerning insight and relationship. Analysts should pay more attention to the similarities between child and adult analysis, so that information can be shared between both subspecialties. PMID- 8717481 TI - Children's developing theory of mind. AB - Studies from the developmental research perspective inform analytic understanding of the neuropsychological preconditions necessary for children's increasing awareness of their own and others' mental lives. Conversely, psychoanalytically informed observations of children's early development demonstrate how the child's emerging understanding of other minds is contextualized in the earliest interactions between parent and child. The notion of a gradually developing understanding of mind that becomes most evident in observational studies between 4 and 6 years of age provides a developmental context for considering children's capacity for internalization, their creation of an inner, psychic reality, and their response to therapeutic interventions during an analytic hour. Significant for psychoanalytic work with children is the suggestion that only after children are able to understand how their own and other's mental states are constructed and how such states are behind all actions and language can they then reflect upon their own thoughts and mental life. Developing a theory of the mind of the other integrates psychoanalytic notions of self-other differentiation, of the development of internalized representations of others, and ultimately of the capacity for self-reflection. PMID- 8717482 TI - Neurosis in childhood and in psychoanalysis: a developmental reformulation. AB - It is widely believed that the transference neurosis, understood as both a repetition of the infantile neurosis, and as an opportunity for new forms of object relations, is the hallmark of psychoanalysis. Because the concept has become caught up in such polemics over the years and has acquired such a variety of meanings, some call for its abandonment; some also call for abandoning the concept of the infantile neurosis. This paper argues that both these concepts refer to significant clinical phenomena and so rather than being abandoned, they should be clarified and redefined in the light of contemporary developmental theory. PMID- 8717483 TI - The transference and the zone of proximal development. AB - The Zone of Proximal Development (ZPD) is discussed, a construct that, when introduced into psychoanalysis, advances understanding of the key clinical relationship between the intrapsychic and the interpersonal. Strands from several psychoanalytic formulations are brought together and forged into a coherent construct, which is then contrasted with the transference. It is shown how the ZPD provides the transference with its mutative potential. Just as the transference provides the motivation for the recruitment of objects to accomplish its purposes (repetition), the ZPD leads to the recruitment of objects in order to accomplish its purposes (to learn by ushering individuals into a speech and internalization community). Under the sway of the transference objects are sought so that early disregulating experiences can be repeated and an opportunity provided for a better resolution. The ZPD works in tandem with the transference, capitalizing on the impetus provided, allowing for the possibility of internalization, a beneficial outcome to transference repetition which otherwise would have no agent of conflict resolution. In analysis, when the transference and the ZPD enjoin smoothly, the potential outcome is "insight" in a broad sense. The processes of the ZPD define the optimal interpersonal context of psychoanalysis, one that allows the intrapsychic to be best reached by analytic interventions. Given the inevitability of mutual influences between analyst and analysand, the analyst strives simultaneously to be in the ZPD yet outside the transference with the analysand, a crucial tension that is a constant, precarious technical factor. This useful tension casts light on such procedural guides as optimal frustration and abstinence. PMID- 8717484 TI - The role of the testicles in male psychological development. AB - In this paper the role of the testicles in male psychological development is thoroughly investigated. All previous psychoanalytic studies are reviewed; new clinical material is presented; critical research data from pediatric urology and physiological sex research are utilized. A number of significant conclusions emerge. (1) Certain cases of pathological castration anxiety are explained by a psychosomatic phenomenon (the "testicular vicious cycle") caused by actual and perceived threats to the genitals. (2) Testicular factors may be decisive in various neurotic symptom formations beginning with the early toilet training period. The testicular contribution then takes place along certain preferred pathways of symbolic displacement. (3) Testicular symptoms are observed in many cases of physical and sexual abuse. This finding has application to the treatment of male abuse survivors and to their related sexual dysfunctions. (4) Masturbation involving the testicles is not infrequent in young boys; the most common practice consists of inserting the testes inside the body. (5) The condition of cryptorchism may contribute to psychopathology, especially in the area of masculine self-image; however, cryptorchid boys do not tend to be gender disordered, effeminate, or prehomosexual. (6) Male body image formation after the phallic stage is a complex emotional and intellectual task involving temporary denial of the inner body and the testicles. Finally it is suggested that a revised account of male sexuality, including both penis and testicles, is a prerequisite for any psychoanalytic theory of gender. PMID- 8717485 TI - Psychoanalytic process: comparison of child, adolescent, and adult analyses. Panel report. PMID- 8717486 TI - Attitudes and experiences of psychoanalysts in analyzing homosexual patients. PMID- 8717488 TI - Relationship between burst properties and sensitivity to input: a theoretical analysis. AB - This paper examines the sensitivity of endogenous bursters to a brief input pulse. The interneurons of the lobster cardiac ganglion were selected as a case study. Using a mathematical model specifically developed for the neurons in the cardiac ganglion of the lobster (Av-Ron et al., 1993), we show a tight link between burst characteristics and certain other parameters. We show that cells with different burst properties differ in their sensitivity to an input of a brief pulse. Irrespective of these differences, all cells display a bimodal response to a brief pulse applied during the quiescent period. During the first three-quarters of the quiescent period, they respond by producing a single spike at most. During the remaining one-quarter, the brief pulse can initiate the cells' intrinsic burst. Our predictions fit experimental results obtained by Tazaki and Cooke (1979). The results obtained herein are discussed with respect to fault tolerance considerations. PMID- 8717487 TI - Chaos and synchrony in a model of a hypercolumn in visual cortex. AB - Neurons in cortical slices emit spikes or bursts of spikes regularly in response to a suprathreshold current injection. This behavior is in marked contrast to the behavior of cortical neurons in vivo, whose response to electrical or sensory input displays a strong degree of irregularity. Correlation measurements show a significant degree of synchrony in the temporal fluctuations of neuronal activities in cortex. We explore the hypothesis that these phenomena are the result of the synchronized chaos generated by the deterministic dynamics of local cortical networks. A model of a "hypercolumn" in the visual cortex is studied. It consists of two populations of neurons, one inhibitory and one excitatory. The dynamics of the neurons is based on a Hodgkin-Huxley type model of excitable voltage-clamped cells with several cellular and synaptic conductances. A slow potassium current is included in the dynamics of the excitatory population to reproduce the observed adaptation of the spike trains emitted by these neurons. The pattern of connectivity has a spatial structure which is correlated with the internal organization of hypercolumns in orientation columns. Numerical simulations of the model show that in an appropriate parameter range, the network settles in a synchronous chaotic state, characterized by a strong temporal variability of the neural activity which is correlated across the hypercolumn. Strong inhibitory feedback is essential for the stabilization of this state. These results show that the cooperative dynamics of large neuronal networks are capable of generating variability and synchrony similar to those observed in cortex. Auto-correlation and cross-correlation functions of neuronal spike trains are computed, and their temporal and spatial features are analyzed. In other parameter regimes, the network exhibits two additional states: synchronized oscillations and an asynchronous state. We use our model to study cortical mechanisms for orientation selectivity. It is shown that in a suitable parameter regime, when the input is not oriented, the network has a continuum of states, each representing an inhomogeneous population activity which is peaked at one of the orientation columns. As a result, when a weakly oriented input stimulates the network, it yields a sharp orientation tuning. The properties of the network in this regime, including the appearance of virtual rotations and broad stimulus dependent cross-correlations, are investigated. The results agree with the predictions of the mean field theory which was previously derived for a simplified model of stochastic, two-state neurons. The relation between the results of the model and experiments in visual cortex are discussed. PMID- 8717490 TI - A simple model of human foveal ganglion cell responses to hyperacuity stimuli. AB - We developed a physiologically plausible model of the first steps of spatial visual information processing in the fovea of the human retina. With the predictions of this model we could support the hypothesis that, for moderate contracts (< or = 40%), hyperacuity is mediated by the magnocellular (MC-) pathway. Despite the lower sampling density in the MC pathway, as compared to the parvocellular (PC-) pathway, the information that is transferred by the MC ganglion cells is sufficient to achieve thresholds comparable to those of human subjects in psychophysical tasks. This is a result of the much higher signal-to noise ratio of the MC pathway cell signals. The PC pathway cells do not transfer enough information for hyperacuity thresholds. PMID- 8717489 TI - Coding of odor intensity in a steady-state deterministic model of an olfactory receptor neuron. AB - The coding of odor intensity by an olfactory receptor neuron model was studied under steady-state stimulation. Our model neuron is an elongated cylinder consisting of the following three components: a sensory dendritic region bearing odorant receptors, a passive region consisting of proximal dendrite and cell body, and an axon. First, analytical solutions are given for the three main physiological responses: (1) odorant-dependent conductance change at the sensory dendrite based on the Michaelis-Menten model, (2) generation and spreading of the receptor potential based on a new solution of the cable equation, and (3) firing frequency based on a Lapicque model. Second, the magnitudes of these responses are analyzed as a function of odorant concentration. Their dependence on chemical, electrical, and geometrical parameters is examined. The only evident gain in magnitude results from the activation-to-conductance conversion. An optimal encoder neuron is presented that suggests that increasing the length of the sensory dendrite beyond about 0.3 space constant does not increase the magnitude of the receptor potential. Third, the sensitivities of the responses are examined as functions of (1) the concentration at half-maximum response, (2) the lower and upper concentrations actually discriminated, and (3) the width of the dynamic range. The overall gain in sensitivity results entirely from the conductance-to-voltage conversion. The maximum conductance at the sensory dendrite appears to be the main tuning constant of the neuron because it determines the shift toward low concentrations and the increase in dynamic range. The dynamic range of the model cannot exceed 5.7 log units, for a sensitivity increase at low odor concentration is compensated by a sensitivity decrease at high odor concentration. PMID- 8717491 TI - Cleavage of structural components of mammalian vitreous by endogenous matrix metalloproteinase-2. AB - Our goal was to determine if the major endogenous vitreous matrix metalloproteinase (MMP-2) could digest known collagenous components of the vitreous body. Matrix metalloproteinase-2 and its associated inhibitors were isolated from porcine vitreous by affinity column chromatography. The inhibitors were inactivated by chemical modification with dithiothreitol and iodoacetamide. The latent MMP-2 was then activated with the organomercurial, p-aminophenyl mercuric acetate (APMA). Bovine vitreous fibrillar collagens (types II, V/XI and IX) were isolated by pepsin extraction and differential salt precipitation. Intact type IX collagen was purified by selective salt precipitation followed by ion exchange and size exclusion chromatography. These isolated collagens were incubated for 6 to 24 h with different concentrations of activated MMP-2, and the extent of collagen degradation was analyzed. Activated MMP-2 was also introduced into freshly isolated vitreous gels and the degree of liquefaction was determined. Our results showed that the activated MMP-2 has no apparent effect upon type II collagen but can degrade type V/XI collagen and type IX collagen fragments (COL2 and COL2 + COL3). In addition, when the type IX collagen was in the intact helical form, MMP-2 appeared to selectively digest alpha 3 (IX) chains. This suggested that vitreous MMP-2 preferentially cleaved certain vitreous collagen chains into large fragments rather than small peptides. MMP-2 also disrupted the vitreous gel in vitro, releasing proteins but not hexuronic acid or sulfated glycosaminoglycans into the liquefied supernatant. We conclude that MMP-2 activity should be considered as a potential mechanism of vitreous liquefaction that is seen in aging and various pathological states. PMID- 8717492 TI - [Functional role and properties of AMP-deaminase]. AB - AMP-deaminase (AMPDA) catalyzes the deamination of AMP to IMP and ammonia. Being an integral enzyme of the purine nucleotide cycle (PNC), AMPDA participates in catalytic deamination of amino acids and provides their involvement in a carbohydrate metabolism, fumarate being one of the end products of PNC. Since AMPDA competes with 5'-nucleotidase for AMP, it is responsible for regulation of a physiologically important active product of purine nucleotide metabolism, such as adenosine. Thus, this enzyme plays an important role in determining the physiological state of the organism in normal conditions as well as under the influence of some environmental factors and in some pathologies. The review sums up the information concerning the AMPDA participation in PNC operation in animal tissues, coding genes and enzyme activity regulation by various effectors, including, reversible phosphorylation and binding to myofibrils and myosin. Special attention is being given to a possible relationship of AMPDA activity deficiency to some neuromuscular pathologies. PMID- 8717493 TI - [Non-native conformational states of immunoglobulins: thermodynamic and functional analysis of rabbit IgG]. AB - By changing pH within the pH range 2-7, the processes of intramolecular rearrangement in the multidomain structure of rabbit IgG were induced which resulted in the formation of four IgG conformers - N, NA, I and NI, differing in the amount of structure in the CH2 domain and as well as in its interaction with the neighboring domains. The stability of the IgG conformers was studied by differential scanning calorimetry and expressed in terms of thermodynamic parameters, delta H, Tm, and delta G. The pH-dependent changes in thermal stability of IgG and the range of stability for the conformers were described by conformational (phase) diagrams. The native N-conformer exists at pH 7.0-5.5. At pH < 5.5 the formation of the NA conformer showing significant decooperation (lower extent of interaction) of the domains occurs. Decooperation permits the CH2 domain in the NA conformer to undergo conformational transitions independently of the rest of the molecule-the property which cannot be observed for the more cooperative N-conformer of IgG. The formation of the intermediate I state occurs at pH < 3 as a consequence of unfolding of the tertiary structure of the CH2 domain, while its secondary structure and compact tertiary structure of the remainder of the IgG domains remain unchanged. Such partially structured and stable states of immunoglobulins have not yet been described. Reversibility of pH induced transitions in IgG was studied using calorimetry and ligand-binding assays involving the C1q component of the complement, protein A, antigen and monospecific anti-CH2 domain antibodies as conformational probes reporting local conformational changes in the CH2 domain and its interdomain interfaces. Refolding of IgG from the I-state by increasing pH results in the formation of a functionally active N1 conformer differing from native IgG by a changed balance of interdomain interactions. In addition to previously reported data suggesting a greater extent of the CH2 and CH1 domain interactions in the NI conformer, this conformer was shown to possess a lower extent of interaction of the CH2 and CH3 domains. The changed stability and a balance of domain interactions in the NI conformer results in changes in some of the effector functions including enhanced two-site binding to protein A and antigen-dependent binding to C1q with a concomitant decrease in the affinity of protein A binding site and reduced antigen binding capacity of the high affinity IgG. Low affinity IgG does not change the antigen-binding parameters after refolding to the N1 conformer. PMID- 8717494 TI - [Effect of plasma lipoproteins and apolipoprotein A-I on lipopolysaccharide induced production of reactive oxygen metabolites by Kupffer cells of the rat]. AB - The effects of plasma lipoproteins on LPS-induced production of reactive oxygen intermediates (ROI) in Kupffer cells isolated from livers of zymosan-treated rats, have been studied. ROI production was low in normal Kupffer cells but increased 18-fold by LPS in Kupffer cells from zymosan-treated rats. However, preincubation of LPS with rat serum, low density lipoproteins, high density lipoproteins (HDL3) and apolipoprotein A-I (apoA-I) decreased the LPS-induced production of ROI by 1.2-, 1.3-, 2.8-, and 1.5-fold, respectively. In contrast, very low density lipoproteins and HDL2 had no significant effect on the LPS induced production of ROI. These results provide evidence for LPS-HDL3 complex formation and suggest that HDL3 and apoA-I play an important role in host defense against endotoxemia and infection. PMID- 8717495 TI - [Energy-dependent processes and metabolism of adenylates in human erythrocytes]. AB - Amphotericin B (1-3 mg/l) decreases the ATP content in erythrocytes by 11-26% and stimulates the K+ efflux but has no effect on the adenylate pool. Adenosine added to the erythrocyte suspension increases the adenylate pool, maintains a high intracellular ATP level for 6-8 hours of incubation at 37 degrees C and diminishes the amphotericin B-induced leakage of K+. Incubation of erythrocytes without glucose for 4-5 hours leads to a 20-50% loss of ATP accompanied by a significant reduction of the adenylate pool. Further addition of glucose partly restores the ATP level. In the presence of adenosine the ATP concentration is restored far more pronounced reaching nearly the original level due to the increase of the adenylate pool. PMID- 8717496 TI - [Spectral studies of the ceruloplasmin active site during its copper repletion and depletion]. AB - Ceruloplasmin (CP; ferro-O2-oxidoreductase, EC 1.16.3.1) was gradually depleted of different type Cu2+ by dialysis against KCN. Protein samples were taken 2, 4, 6, 22 and 28 hrs after the beginning of dialysis. The number of copper atoms per CP molecule was estimated using atomic absorption spectrophotometry. Light absorption in UV and visible regions, fluorescence and EPR spectra were also recorded. The experimental results allowed to trace the sequence of release of certain Cu2+ ions from the CP molecule. The same methods used in the course of CP active center reconstitution made it possible to determine the dependence of its repletion with certain Cu2+ on the degree of the preceding catalytic center destruction. It was shown that efficient recurrence of the CP oxidase activity is achieved if after dialysis against KCN the enzyme retains type III Cu2+ in its active center. The data obtained allow to specify more precisely the role of different types of Cu2+ in the assembly of the complex catalytic center of CP and in exercising by the enzyme of its multiple functions. PMID- 8717498 TI - [Effect of degree of aerosol OT reverse micelle hydration on the catalytic activity of peroxidase, its conjugate with cortisol and their immune complexes]. AB - In Aerosol OT (AOT) reversed micelles in heptane, the oxidation kinetics of ortho phenylenediamine (PDA) catalyzed by peroxidase (HRP), its conjugate with nine cortisol molecules and their immunocomplexes with monoclonal antibodies AT-2C specific to domain I of HRP has been studied with regard to the hydration degree of micelles W(0) = [H2O]/[A0T] and the concentration of solubilized biocatalysts. The profiles of dependencies of the initial PDA oxidation rate, v(0), on W(0) varied with an increase in the concentrations of HRP, HRP-cortisol and their immunocomplexes in micellar systems. The rise in HRP concentration from 0.5 up to 1.5 nM increases the v(0) values at a constant hydration degree, W(0) and is characterized by a limiting value of the initial rate at W(0) > 30. At HRP cortisol concentrations of 1.2 and 1.5 nM, the dependences v(0)-W(0) have three maxima with gradually increasing values of the initial rate which may be accounted for by self-association of the hydrophobized conjugate, HRP-cortisol, in AOT micelles. The profiles of dependencies, v(0)-W(0) for HRP immunocomplexes (HRP-cortisol) with monoclonal antibodies, AT-2C, at various relationships of the components are characterized by three maxima with increasing values of v(0) which may by related to initial biocatalysts (HRP or HRP-cortisol) and their immunocomplexes with one or two molecules of AT-2C. PMID- 8717497 TI - [Ferritin--a biocatalyst of aromatic amine oxidation]. AB - The tetramethylbenzidine (TMB) oxidation kinetics dependent on spleen ferritin (FERR) has been studied in TMB-FERR-H2O2 (1) and TMB-FERR-O2 (2) systems at pH 4.2 and 6.0. At TMB concentrations below 6 mM, the dependencies of the initial rate for protein oxidation in system (1) on initial concentrations of TMB and H2O2 are described by the Michaelis-Menten kinetics. In terms of (kcat/K(m)), the maximum FERR efficiency during TMB oxidation is equal to 2.8-10(-1) s-1. Mannitol, urea, ethanol and sodium fluoride strongly inhibit TMB oxidation in system (1). Competitive type oxidation inhibition by urea is characterized by K(i) = 12.5 mM. At TMB concentrations above 6 mM in system (2) the autocatalytic oxidation of the amine is observed. In both systems the initial rate of TMB oxidation increases strongly with a decrease in pH from 9.3 down to 4.2. In system (1) the initiating role is played by Fe3+ ions interaction with H2O2, that in system (2)-by the interaction of the same ions with the amine proper. In both cases, TMB oxidation is a catalytic chain process including elementary ion radical stages. The main oxidizing agents in system (2) are HO radicals and Fe3+ ions. The TMB interaction with FERR may be the reason for the lipid peroxidation increase following administration of aromatic amines. PMID- 8717499 TI - [Proteolytic enzymes from Streptomyces fradiae: a metalloendopeptidase, subtilisin-like, and trypsin-like proteinases]. AB - Three proteolytic enzymes-the metalloproteinase, SFMP, and two serine proteinases, SFSP and SFTP-have been isolated and purified from the culture fluid of Streptomyces fradiae using chromatography on bacitracin-silochrome, bacitracin Sepharose, DEAE-cellulose and fractionation by ammonium sulfate. Study of physico chemical and functional properties of the enzymes and structural analysis revealed that SFMP is a cysteine-containing metalloendopeptidase with M(r) of 36 kDa, has a peak activity for synthetic substrates at pH 7.0-7.5 and at 60-65 degrees C and is stable at pH 7.0-9.0. The serine proteinase SFSP is related to subtilisin-like enzymes, has a M(r) of 29 kDa and a pH optimum at 7.5-8.5 at temperature up to 50 degrees C. The proteinase is stable at pH 4.0-9.0 and retains 30% of its activity at 70 degrees C. The other serine proteinase, SFTP, has a M(r) of 26 kDa and is related to trypsin-like enzymes. Its activity for synthetic substrates of trypsin is maximal at pH 6.8-8.8 at 50 degrees C. The enzyme is stable at pH 4.5-8.5 and at temperature below 50 degrees C. It has been shown that Streptomyces fradiae, like Streptomyces griseus and other Streptomycetes, possesses an ability to secrete serine proteinases (SFSP and SFTP) related to two evolutionally distinct families of serine proteinases, i.e., subtilisin and chymotrypsin families. SFMP and SFSP have been isolated and characterized for the first time. PMID- 8717500 TI - [Bacillus cereus chitinases: isolation and characteristics]. AB - Three chitinases (M(r) = 68, 52 and 38 kDa) have been isolated from the cultural filtrate of Bacillus cereus strain VKPM B-6838 by stepwise hydrophobic chromatography on butyl-Toyopearl and gel filtration on Superdex 75 (FPLC). The chitinases are stable in the pH range 4-10 and have the same pH optimum of activity. The 68 and 38 kDa enzymes display the highest activity at 60 degrees C. while the 52 kDa chitinase-at 50 degrees C. In contrast with the 68- and 52 kDa enzymes, the 38 kDa chitinase hydrolyzes not only colloidal but also "crystalline" chitin and chitosan. None of the chitinases hydrolyzes chitobiose. The N-terminal sequences (10 amino acids) of the 52 and 38 kDa chitinases do not reveal structural similarity between themselves or to other known bacterial chitinases. The 68 kDa chitinase is not immunologically related to the 52 and 38 kDa enzymes. The results obtained suggest that the 68, 52 and 38 kDa chitinase are the unique proteins. PMID- 8717501 TI - [Gnathologic concepts and their practical role in dentistry]. PMID- 8717502 TI - [Familial case of Papillon-Lefevre syndrome]. PMID- 8717503 TI - [Hypnotic desensitization as a supplemental method in dental care of patients with panic disorder. Report of a case]. PMID- 8717504 TI - Report of the National Commission on Allied Health. PMID- 8717505 TI - Altered peptide ligand-induced partial T cell activation: molecular mechanisms and role in T cell biology. AB - The elucidation of the phenomena of T cell antagonism and partial activation by altered peptide ligands has necessitated a revision in the traditional concepts of TCR recognition of antigen and subsequent signal transduction. Whereas previous models supported a single ligand specificity for any particular T cell, many studies using analogs of immunogenic peptides have now demonstrated a flexibility in this recognition. Moreover, interaction with such altered peptide ligands can result in dramatically different phenotypes of the T cells, ranging from inducing selective stimulatory functions to completely turning off their functional capacity. Investigations of the biochemical basis leading to these phenotypes have shown that altered peptide ligands can induce a qualitatively different pattern of signal transduction events than does any concentration of the native ligand. Such observations imply that several signaling modules are directly linked to the TCR/CD3 complex and that they can be dissociated from each other as a direct result of the nature of the ligand bound. Interestingly, many in vivo models of T cell activation are compatible with a selective signaling model, and several studies have shown that peptide analogs can play a role in various T cell biologic phenomena. These data strongly suggest that naturally occurring altered peptide ligands for any TCR exist in the repertoire of self peptides or, in nature, derived from pathogens, and recent reports provide compelling evidence that this is indeed the case. The concept of altered peptide ligands, their effects on T cell signaling, the hypothesized mechanisms by which they exert their effects, and their possible roles in shaping the T cell immune response are the scope of this review. PMID- 8717506 TI - Early T lymphocyte progenitors. AB - The earliest steps along the pathway leading to T cells in mice and humans are reviewed. These are the steps between the multipotent hemopoietic stem cell (HSC) and the fully committed precursors undergoing T cell receptor (TCR) gene rearrangement. At this level significant differences between adult and fetal lymphopoiesis have been demonstrated. The extent of lymphoid commitment of precursors within bone marrow is still unresolved, although HSCs clearly undergo developmental changes before migration to the thymus. Both multipotent and T restricted precursors have now been isolated from fetal blood, suggesting both may seed the thymus. Within the thymus, several minute but discrete populations of T precursors precede the stage of TCR gene rearrangement. They include precursors that are not exclusively T-lineage committed, although they are distinct from HSCs. These precursors have a potential to form NK cells, B cells, dendritic cells, and sometimes other myeloid cells. Some factors that control early lymphoid development are discussed, including IL-7 and the Ikaros transcription factors. These will eventually help to clarify the process of T lineage commitment. PMID- 8717507 TI - Immunotoxins: an update. AB - The use of immunotoxins (ITs) in the therapy of cancer, graft-vs-host disease (GvHD), autoimmune diseases, and AIDS has been ongoing for the past two decades. ITs contain a targeting moiety for delivery and a toxic moiety for cytotoxicity. Theoretically, one molecule of a toxin, routed to the appropriate cellular compartment, will be lethal to a cell. Newly developed MoAbs, toxins, and molecular biological technologies have enabled researchers to construct ITs that can effectively kill many different cell types. In fact, phase I/II clinical trials have given promising results. Although nonspecific toxicity and immunogenicity still limit the use of IT therapy, these agents hold enormous promise in an optimal setting to treat minimal disease. PMID- 8717508 TI - Cellular interactions in thymocyte development. AB - Interactions between stromal cells and thymocytes play a crucial role in T cell development. The thymic stroma is complex and consists of epithelial cells derived from the pharyngeal region during development, together with macrophages and dendritic cells of bone marrow origin. In addition, fibroblasts and matrix molecules permeate the whole framework. It is now apparent that these individual stromal components play specialized roles at different stages of T cell differentiation. Thus, at the early CD4-8- stage of development, T cell precursors require fibroblast as well as epithelial cell interactions. Later, at the CD4+8+ stage, as well as providing low avidity TCR/MHC-peptide interactions, thymic epithelial cells have been shown to possess unique properties essential for positive selection. Dendritic cells, on the other hand, are probably efficient mediators of negative selection, but they may not be solely responsible for this activity. Alongside the functional roles of stromal cells, considerable progress is being made in unraveling the nature of the signaling pathways involved in T cell development. Identification of the pre-T cell receptor (pre TCR) and associated signaling molecules marks an important advance in understanding the mechanisms that control gene rearrangement and allelic exclusion. In addition, a better understanding of the signaling pathways that lead to positive selection on the one hand and negative selection on the other is beginning to emerge. Many issues remain unresolved, and some are discussed in this review. What, for example, is the nature of the chemotactic factor(s) that attract stem cells to the thymus? What is the molecular basis of the essential interactions between early thymocytes and fibroblasts, and early thymocytes and epithelial cells? What is special about cortical epithelial cells in supporting positive selection? These and other issues are ripe for analysis and can now be approached using a combination of modern molecular and cellular techniques. PMID- 8717509 TI - One step ahead of the game: viral immunomodulatory molecules. AB - For decades cell biologists have relied on viruses to facilitate the study of complex cellular function. More recently, the tragedy of the AIDS epidemic has focused considerable human and financial resources on both virology and immunology, resulting in the generation of new information relating these disciplines. As the miracle of the mammalian immune system unfolds in the laboratory, the elegance of the mechanisms used by co-evolving viruses to circumvent detection and destruction by the host becomes inescapably obvious. Although many observation of virus-induced phenomena that likely contribute to the virus's escape of immune surveillance are still empirical, many other such phenomena have now been defined at the molecular level and confirmed in in vivo models. Immune modulators encoded within viral genomes include proteins that regulate antigen presentation, function as cytokines or cytokine antagonists, inhibit apoptosis, and interrupt the complement cascade. The identification of such gene products and the elucidation of their function have substantially strengthened our understanding of specific virus-host interactions and, unexpectedly, have contributed to the recognition of potent synergy between viruses, which can result in an unpredictable exacerbation of disease in co infected individuals. Because many viral immune modulators clearly have host counterparts, viruses provide a valuable method for studying normal immune mechanisms. It is conceivable that an improved understanding of virus-encoded immunomodulators will enhance our ability to design reagents for use in therapeutic intervention in disease and in vaccine development. PMID- 8717510 TI - Genetic analysis of tyrosine kinase function in B cell development. AB - B lymphopoiesis is regulated by multiple signals from stromal cell contact, soluble cytokines, antigen, and T helper cells. In vitro and biochemical experiments have implicated tyrosine kinases as key components of many of these signaling pathways. Genetic analysis of the role of these tyrosine kinases has been facilitated by recent advance in transgenic and gene targeting technology as well as by the identification of the genetic basis of several human and murine immune deficiencies. This review discusses the effect of gain and loss of function mutations of selected tyrosine kinases and their regulators and substrates on B cell development and function. PMID- 8717511 TI - Orchestrated information transfer underlying leukocyte endothelial interactions. AB - The specificity and efficiency of leukocyte binding to endothelial cells (ECs) depends on coordinated information transfer from the underlying tissue to endothelium and from there to the leukocyte. We address three distinct information-transfer points in this system: 1, How does the leukocyte read information from the EC? This process is best accounted for by the paradigm of a multi-step adhesion cascade optimized for rapid information readout; it consists of primary adhesion (rolling/tethering), triggering, and strong adhesion. Recent studies with T cells, monocytes, and eosinophils confirm the generality of the paradigm. The concept of primary adhesion has been expanded to involve not only the selectins, but also certain integrins; furthermore, it depends on receptor concentration on leukocyte microvilli. 2. What information from the underlying tissue does the EC transform into signals for the leukocytes? And what rules govern that process? We illustrate the principles with chemokines, believed to participate in the triggering step. The endothelium displays chemokines either (a) directly by "posting" them from other cells or (b) by integrating a variety of tissue and environmental signals and "relaying" that information by producing its own chemokines and surface adhesion molecules. The rules for this endothelial transduction include specificity coupled with redundancy, amplification, synergy, and coordinated induction of ensembles of molecules. Finally, 3. How does the relevant information reach the endothelium? Simple diffusion is sufficient to deliver signals from cells close to the vessel. However, longer range soluble mediator transport appears to be facilitated by fiber bundles, particularly those ensheathed by fibroblastic reticular cells in the lymph node. PMID- 8717512 TI - The interleukin-2 receptor gamma chain: its role in the multiple cytokine receptor complexes and T cell development in XSCID. AB - Interleukin 2 (IL-2), a T cell-derived cytokine, targets a variety of cells to induce their growth, differentiation, and functional activation. IL-2 inserts signals into the cells through IL-2 receptors expressed on cell surfaces to induce such actions. In humans, the functional IL-2 receptor consists of the subunit complexes of the alpha, beta and gamma chains, or the beta and gamma chains. The third component, the gamma chain, of IL-2 receptor plays a pivotal role in formation of the full-fledged IL-2 receptor, together with the beta chain, the gamma chain participates in increasing the IL-2 binding affinity and intracellular signal transduction. Moreover, the cytokine receptors for at least IL-2, IL-4, IL-7, IL-9, and IL-15 utilize the same gamma chain as an essential subunit. Interestingly, mutations of the gamma chain gene cause human X-linked severe combined immunodeficiency (XSCID) characterized by a complete or profound T cell defect. Among the cytokines sharing the gamma chain, at least IL-7 is essentially involved in early T cell development in the mouse organ culture system. The molecular identification of the gamma chain brought a grasp of the structures and functions of the cytokine receptor and an in-depth understanding of the cause of human XSCID. To investigate the mechanism of XSCID and development of gene therapy for XSCID, knockout mice for the gamma chain gene were produced that showed similar but not exactly the same phenotypes as human XSCID. PMID- 8717513 TI - Molecular mechanisms of lymphocyte-mediated cytotoxicity and their role in immunological protection and pathogenesis in vivo. AB - Studies with perforin-deficient mice have demonstrated that two independent mechanisms account for T cell-mediated cytotoxicity: A main pathway is mediated by the secretion of the pore-forming protein perforin by the cytotoxic T cell, whereas an alternative nonsecretory pathway relies on the interaction of the Fas ligand that is upregulated during T cell activation with the apoptosis-inducing Fas molecule on the target cell. NK cells use the former pathway exclusively. The protective role of the perforin-dependent pathway has been shown for infection with the noncytopathic lymphocytic choriomeningitis virus, for infection with Listeria monocytogenes, and for the elimination of tumor cells by T cells and NK cells. In contrast, perforin-dependent cytotoxicity is not involved in protection against the cytopathic vaccinia virus and vesicular stomatitis virus. LCMV induced immunopathology and autoimmune diabetes have been found to require perforin-expression. A contribution of perforin-dependent cytotoxicity to the rejection of MHC class I-disparate heart grafts has also been observed. Its absence is efficiently compensated in rejection of fully allogeneic organ or skin grafts. So far, evidence for a role of Fas-dependent cytotoxicity as a T cell effector mechanism in vivo is lacking. Current data suggest that the main function of Fas may be in regulation of the immune response and apparently less at the level of an effector mechanism in host defense. Further analysis is necessary, however, to settle this point finally. PMID- 8717514 TI - CD28/B7 system of T cell costimulation. AB - T cells play a central role in the initiation and regulation of the immune response to antigen. Both the engagement of the TCR with MHC/Ag and a second signal are needed for the complete activation of the T cell. The CD28/B7 receptor/ligand system is one of the dominant costimulatory pathways. Interruption of this signaling pathway with CD28 antagonists not only results in the suppression of the immune response, but in some cases induces antigen specific tolerance. However, the CD28/B7 system is increasingly complex due to the identification of multiple receptors and ligands with positive and negative signaling activities. This review summarizes the state of CD28/B7 immunobiology both in vitro and in vivo; summarizes the many experiments that have led to our current understanding of the participants in this complex receptor/ligand system; and illustrates the current models for CD28/B7-mediated T cell and B cell regulation. It is our hope and expectation that this review will provoke additional research that will unravel this important, yet complex, signaling pathway. PMID- 8717515 TI - T cell antigen receptor signal transduction pathways. AB - The T cell antigen receptor (TCR) regulates the activation and growth of T lymphocytes. The initial membrane proximal event triggered by the TCR is activation of protein tyrosine kinases with the resultant phosphorylation of cellular proteins. This biochemical response couples the TCR to a divergent array of signal transduction molecules including enzymes that regulate lipid metabolism, GTP binding proteins, serine/threonine kinases, and adapter molecules. The ultimate aim of studies of intracellular signaling mechanisms is to understand the functional consequences of a particular biochemical event for receptor function. The control of cytokine gene expression is one of the mechanism that allows the TCR to control immune responses. Accordingly, one object of the present review is to discuss the role of the different TCR signal transduction pathways in linking the TCR to nuclear targets: the transcription factors that control the expression of cytokine genes. PMID- 8717516 TI - Antigen sampling across epithelial barriers and induction of mucosal immune responses. AB - Epithelial barriers on mucosal surfaces at different sites in the body differ dramatically in their cellular organization, and antigen sampling strategies at diverse mucosal sites are adapted accordingly. In stratified and pseudostratified epithelia, dendritic cells migrate to the outer limit of the epithelium, where they sample antigens for subsequent presentation in local or distant organized lymphoid tissues. In simple epithelia, specialized epithelial M cells (a phenotype that occurs only in the epithelium over organized lymphoid follicles) deliver samples of foreign material by transepithelial transport from the lumen to organized lymphoid tissues within the mucosa. Certain pathogens exploit the M cell transport process to cross the epithelial barrier and invade the mucosa. Here we review the features of M cells that determine antigen and pathogen adherence and transport into mucosal lymphoid tissues. PMID- 8717517 TI - Regulation of MHC class II genes: lessons from a disease. AB - Precise regulation of major histocompatibility complex class II (MHC-II) gene expression plays a crucial role in the control of the immune response. A major breakthrough in the elucidation of the molecular mechanisms involved in MHC-II regulation has recently come from the study of patients that suffer from a primary immunodeficiency resulting from regulatory defects in MHC-II expression. A genetic complementation cloning approach has led to the isolation of CIITA and RFX5, two essential MHC-II gene transactivators. CIITA and RFX5 are mutated in these patients, and the wild-type genes are capable of correcting their defect in MHC-II expression. The identification of these regulatory factors has furthered our understanding of the molecular mechanisms that regulate MHC-II genes. CIITA was found to be a non-DNA binding transactivator that functions as a molecular switch controlling both constitutive and inducible MHC-II expression. The finding that RFX5 is a subunit of the nuclear RFX-complex has confirmed that a deficiency in the binding of this complex is indeed the molecular basis for MHC-II deficiency in the majority of patients. Furthermore, the study of RFX has demonstrated that MHC-II promoter activity is dependent on the binding of higher order complexes that are formed by highly specific cooperative binding interactions between certain MHC-II promoter-binding proteins. Two of these proteins belong to families of which the other members, although capable of binding to the same DNA motifs, are probably not directly involved in the control of MHC-II expression. Finally, the facts that CIITA and RFX5 are both essential and highly specific for MHC-II genes make possible novel strategies designed to achieve immunomodulation via transcriptional intervention. PMID- 8717518 TI - On immunological memory. AB - Immunological memory is a hallmark of the immune system. Evolution can teach us which effector arms of immunological memory are biologically relevant against which virus. Antibodies appear to be the critical protective mechanism against cytopathic viruses. Since these viruses cause cell damage and disease directly, particularly in the absence of an immune response, mothers protect their offspring during a critical immunoincompetent period (a consequence of MHC- restricted T cell recognition) by passive transfer of neutralizing antibodies. In contrast, CTL appear to be the crucial effector mechanism against noncytopathic viruses. Since MHC polymorphism has made vertical transmission of T cell memory impossible, immunoincompetent offspring are not, and need not be, protected against such noncytopathic viruses. During the primary response and again during secondary infection, the most important function of CTL is to eliminate noncytopathic viruses, which may otherwise cause lethal immunopathology. Increased precursor frequencies of B and T cells appear to remain in the host independent of antigen persistence. However, in order to protect against cytopathic viruses, memory B cells have to produce antibody to maintain protective elevated levels of antibody; B cell differentiation into plasma cells is driven by persisting antigen. Similarly, to protect against infection with a noncytopathic virus, CTL have to recirculate through peripheral organs. Activation and capacity to emigrate into solid tissues as well as cytolytic effector function are also dependent upon, and driven by, persisting antigen. Because no convincing evidence is available yet of the existence of identifiable B or T cells with specialized memory characteristics, the phenotype of immunological memory correlates best with antigen-driven activation of low frequency effector T cells and plasma cells. PMID- 8717519 TI - Antigen processing and presentation by the class I major histocompatibility complex. AB - Major histocompatibility complex (MHC) class I molecules bind peptides derived from cellular proteins and display them for surveillance by the immune system. These peptide-binding molecules are composed of a heavy chain, containing an antigen-binding groove, which is tightly associated with a light chain (beta 2 microglobulin). The majority of presented peptides are generated by degradation of proteins in the cytoplasm, in many cases by a large multicatalytic proteolytic particle, the proteasome. Two beta-subunits of the proteasome, LMP2 and LMP7, are inducible by interferon-gamma and alter the catalytic activities of this particle, enhancing the presentation of at least some antigens. After production of the peptide in the cytosol, it is transported across the endoplasmic reticulum (ER) membrane in an ATP-dependent manner by TAP (transporter associated with antigen presentation), a member of the ATP-binding cassette family of transport proteins. There are minor pathways for generating presented peptides directly in the ER, and some evidence suggests that peptides may be further trimmed in this location. The class I heavy chain and beta 2-microglobulin are cotranslationally translocated into the endoplasmic reticulum where their assembly may be facilitated by the sequential association of the heavy chain with chaperone proteins BiP and calnexin. The class I molecule then associates with the lumenal face of TAP where it is retained, presumably awaiting a peptide. After the class I molecule binds a peptide, it is released for exocytosis to the cell surface where cytotoxic T lymphocytes examine it for peptides derived from foreign proteins. PMID- 8717520 TI - Role of cytokines in rheumatoid arthritis. AB - Analysis of cytokine mRNA and protein in rheumatoid arthritis tissue revealed that many proinflammatory cytokines such as TNF alpha, IL-1, IL-6, GM-CSF, and chemokines such as IL-8 are abundant in all patients regardless of therapy. This is compensated to some degree by the increased production of anti-inflammatory cytokines such as IL-10 and TGF beta and cytokine inhibitors such as IL-1ra and soluble TNF-R. However, this upregulation in homeostatic regulatory mechanisms is not sufficient as these are unable to neutralize all the TNF alpha and IL-1 produced. In rheumatoid joint cell cultures that spontaneously produce IL-1, TNF alpha was the major dominant regulator of IL-1. Subsequently, other proinflammatory cytokines were also inhibited if TNF alpha was neutralized, leading to the new concept that the proinflammatory cytokines were linked in a network with TNF alpha at its apex. This led to the hypothesis that TNF alpha was of major importance in rheumatoid arthritis and was a therapeutic target. This hypothesis has been successfully tested in animal models, of, for example, collagen-induced arthritis, and these studies have provided the rationale for clinical trials of anti-TNF alpha therapy in patients with long-standing rheumatoid arthritis. Several clinical trials using a chimeric anti-TNF alpha antibody have shown marked clinical benefit, verifying the hypothesis that TNF alpha is of major importance in rheumatoid arthritis. Retreatment studies have also shown benefit in repeated relapses, indicating that the disease remains TNF alpha dependent. Overall these studies demonstrate that analysis of cytokine expression and regulation may yield effective therapeutic targets in inflammatory disease. PMID- 8717521 TI - Accessibility control of antigen-receptor variable-region gene assembly: role of cis-acting elements. AB - Antigen receptor variable region genes are assembled from germline variable (V), diversity (D), and joining (J) gene segments. This process requires expression of V(D)J recombinase activity, and "accessibility" of variable gene segments to this recombinase. The exact mechanism by which variable gene segments become accessible during development is not known. However, several studies have shown that cis-acting elements that regulate transcription may also function to regulate accessibility. Here we review the evidence that transcriptional promoters, enhancers, and silencers are involved in regulation of accessibility. The manner in which these elements may combine to regulate accessibility is addressed. In addition, current and potential strategies for identifying and analyzing cis-acting elements that mediate locus accessibility are discussed. PMID- 8717522 TI - Immunopharmacology of rapamycin. AB - The potent immunosuppressive drugs FK506 and rapamycin interfere with signal transduction pathways required for T cell activation and growth. The distinct inhibitory effects of these drugs on the T cell activation program are mediated through the formation of pharmacologically active complexes with members of a family of intracellular receptors termed the FK506 binding proteins (FKBPs). The FKBP12.FK506 complex specifically binds to and inhibits calcineurin, a signaling protein required for transcriptional activation of the interleukin (IL)-2 gene in response to T cell antigen receptor engagement. The FKBP12. rapamycin complex interacts with a recently defined target protein termed the mammalian target of rapamycin (mTOR). Accumulating data suggest that mTOR functions in a previously unrecognized signal transduction pathway required for the progression of IL-2 stimulated T cells from G1 into the S phase of the cell cycle. Here we review the immunopharmacology of rapamycin, with particular emphasis on the characterization of mTOR. PMID- 8717523 TI - Recognition by gamma/delta T cells. AB - In contrast with the study of alpha beta T cells, that of gamma delta T cells is relatively recent and stems from the discovery of their rearranged genes, rather than from any knowledge of their biological function. Thus, experiments designed to characterize their specificity and function have drawn heavily on our knowledge of alpha beta T cells. During the past few years, many studies, especially with mice lacking either alpha beta or gamma delta T cells, have demonstrated that gamma delta T cells can contribute to immune competence, but they do so in a way that is distinct from alpha beta T cells. It is also evident that gamma delta T cells may not recognize antigen the same way as do alpha beta T cells. Analysis of three protein antigens-the murine MHC class II IEk, the nonclassical MHC T10/T22, and the Herpes virus glycoprotein gI-indicates that gamma delta T cell recognition does not require antigen processing and that the proteins are recognized directly. In all three cases, recognition by these T cell clones involves neither peptides bound to these proteins nor peptides derived from them. Moreover, a group of small phosphate-containing nonpeptide compounds derived from mycobacterial extracts has been found to stimulate a major population of human peripheral gamma delta T cells in a T cell receptor (TCR) dependent manner. This indicates that gamma delta T cells can respond to ligands that are different from those of alpha beta T cells. Analysis of complementarity determining region (CDR3) length distributions of gamma and delta chains indicates that they are more similar to those of immunoglobulins than to TCR alpha and beta. This further supports the idea that gamma delta and alpha beta T cells recognize antigens differently and suggests that gamma delta T cells may be more like immunoglobulins in their recognition properties. gamma delta T cells share many cell surface proteins with alpha beta T cells and are able to secrete lymphokines and express cytolytic activities in response to antigenic stimulation. These, together with the results cited above, indicate that gamma delta T cells can mediate cellular immune functions without a requirement for antigen processing. Thus, pathogens, damaged tissues, or even B and T cells can be recognized directly, and cellular immune responses can be initiated without a requirement for antigen degradation or specialized antigen-presenting cells. This would give gamma delta T cells greater flexibility than the more classical type of alpha beta T cell-mediated cellular immunity. PMID- 8717524 TI - Salmonellosis: host immune responses and bacterial virulence determinants. AB - The lifestyle of bacterial pathogens requires them to establish infection in the face of host immunity. Upon entering a potential host, a variety of interactions are initiated, the outcome of which depends upon a myriad of attributes of each of the participants. In this review we discuss the interactions that occur between pathogenic Salmonella species and the host immune systems, but when appropriate to broaden perspective, we have provided a general overview of the interactions between bacterial pathogens and animal hosts. Pathogenic Salmonella species possess an array of invasion genes that produce proteins secreted by a specialized type III secretion apparatus. These proteins are used by the bacteria to penetrate the intestinal mucosa by invading and destroying specialized epithelial M cells of the Peyer's patches. This maneuver deposits the bacteria directly within the confines of the reticuloendothelial system. The host responds to these actions with nonspecific phagocytic cells and an inflammatory response as well as by activating specific cellular and humoral immune responses. Salmonella responds to this show of force directly. It appears that the bacteria invade and establish a niche within the very cells that have been sent to destroy them. Efforts are underway to characterize the factors that allow these intracellular bacteria to customize intracellular vacuoles for their own purposes. It is the constant play between these interactions that determines the outcome of the host infection, and clearly they will also shape the evolution of new survival strategies for both the bacterium and the host. PMID- 8717525 TI - The structure of the T cell antigen receptor. AB - Recent crystallographic studies of T cell antigen receptor (TCR) fragments from the alpha and beta chains have now confirmed the expected structural similarity to corresponding immunoglobulin domains. Although the three-dimensional structure of a complete TCR alpha beta heterodimer has not yet been determined, these results support the view that the extracellular region should resemble an immunoglobulin Fab fragment with the antigen-binding site formed from peptide loops homologous to immunoglobulin complementarity-determining regions (CDR). These preliminary results suggest that CDR1 and CDR2 may be less variable in structure than their immunoglobulin counterparts, consistent with the idea that they may interact preferentially with the less polymorphic regions of the molecules of the major histocompatibility complex. The region on the variable beta domain responsible for superantigen recognition is analyzed in detail. The implications for T cell activation from the interactions observed between domains of the alpha and beta chains are also discussed in terms of possible dimerization and allosteric mechanisms. PMID- 8717526 TI - Immune regulation by CD40 and its ligand GP39. AB - Over the past three years, CD40 and its ligand (gp39, CD40L, TBAM) have been shown to be essential for humoral immune responses to thymus-dependent antigens. However, as the tissue distribution widens for those cells that express CD40 and gp39, we can now show that this ligand-receptor pair also plays an important role in the selection of self-reactive T cells in the thymus (central tolerance) and the regulation of tolerance in mature T cells (peripheral tolerance). Advances in our understanding of the molecular basis for CD40 biology is based in two areas of research. First, a major breakthrough in our understanding of how CD40 transduces biological events centers on the identification of a novel protein that binds to the cytoplasmic tail of CD40 and may act as a signal transducing molecule. Secondly, advances in molecular modeling and mutagenesis of this ligand receptor pair have helped to identify the critical receptor/ligand contacts in the gp39/CD40 complex. Advances in each of these areas are discussed. PMID- 8717527 TI - Receptors for HLA class-I molecules in human natural killer cells. AB - Natural killer cells are likely to play an important role in the host defenses because they kill virally infected or tumor cells but spare normal self-cells. The molecular mechanism that explains why NK cells do not kill indiscriminately has recently been elucidated. It is due to several specialized receptors that recognize major histocompatibility complex (MHC) class I molecules expressed on normal cells. The lack of expression of one or more class I alleles leads to NK mediated target cell lysis. During NK cell development, the class I-specific receptors have adapted to self-class I molecules on which they recognize epitopes shared by groups of class I alleles. As such, they may fail to recognize either self-molecules that bound unusual peptides or allogeneic class I molecules unrelated to self-alleles. Different types of receptors specific for groups of HLA-C or HLA-B alleles have been identified. While in most instances, they function as inhibiting receptors, an activating form of the HLA-C-specific receptors has been identified in some donors. Molecular cloning of HLA-C- and HLA B-specific receptors has revealed new members of the immunoglobulin superfamily with two or three Ig-like domains, respectively, in their extracellular portion. While the inhibiting form is characterized by a long cytoplasmic tail associated with a nonpolar transmembrane portion, the activating one has a short tail associated with a Lys-containing transmembrane portion. Thus, these human NK receptors are different from the murine Ly49 that is a type II transmembrane protein characterized by a C type lectin domain. A subset of cytolytic T lymphocytes expresses NK-type class I-specific receptors. These receptors exert an inhibiting activity on T cell receptor-mediated functions and offer a valuable model to analyze the regulatory mechanisms involved in receptor-mediated cell activation and inactivation. PMID- 8717528 TI - The NF-kappa B and I kappa B proteins: new discoveries and insights. AB - The transcription factor NF-kappa B has attracted widespread attention among researchers in many fields based on the following: its unusual and rapid regulation, the wide range of genes that it controls, its central role in immunological processes, the complexity of its subunits, and its apparent involvement in several diseases. A primary level of control for NF-kappa B is through interactions with an inhibitor protein called I kappa B. Recent evidence confirms the existence of multiple forms of I kappa B that appear to regulate NF kappa B by distinct mechanisms. NF-kappa B can be activated by exposure of cells to LPS or inflammatory cytokines such as TNF or IL-1, viral infection or expression of certain viral gene products, UV irradiation, B or T cell activation, and by other physiological and nonphysiological stimuli. Activation of NF-kappa B to move into the nucleus is controlled by the targeted phosphorylation and subsequent degradation of I kappa B. Exciting new research has elaborated several important and unexpected findings that explain mechanisms involved in the activation of NF-kappa B. In the nucleus, NF-kappa B dimers bind to target DNA elements and activate transcription of genes encoding proteins involved with immune or inflammation responses and with cell growth control. Recent data provide evidence that NF-kappa B is constitutively active in several cell types, potentially playing unexpected roles in regulation of gene expression. In addition to advances in describing the mechanisms of NF-kappa B activation, excitement in NF-kappa B research has been generated by the first report of a crystal structure for one form of NF-kappa B, the first gene knockout studies for different forms of NF-kB and of I kappa B, and the implications for therapies of diseases thought to involve the inappropriate activation of NF-kappa B. PMID- 8717529 TI - Inhibitory effects of bisphosphonate (YM175) on bone resorption induced by a metastatic bone tumor. AB - The effects of a third-generation bisphosphonate, YM175 (disodium dihydrogen (cycloheptylamino)-methylene-1,1-bisphosphonate), on bone resorption induced by a metastatic human melanoma cell line (A375) were investigated morphologically using an experimental model of bone metastases in nude mice. An injection of A375 in the left cardiac ventricle produced multiple osteolytic lesions. Then, 4 weeks after the cell injection, we administrated YM175 (1 mg/kg) intravenously once and sacrificed the animals 3 days later. On histochemical observation, there was a layer of stromal cells with numerous mononuclear and multinucleated tartrate resistant acid phosphatase (TRAPase)-positive cells in the untreated control group. In contrast, this layer was extensively reduced in most areas, and only a few TRAPase-positive cells were seen around tumor nests and on the bone surface in the experimental group. Most of the TRAPase-positive cells were stained only weakly and/or homogeneously, and there was little evidence of cell polarity. Some of them were vacuolated. Ultrastructurally, they were round and devoid of ruffled borders and clear zones. The findings suggest that YM175 decreases the number and activity of osteoclasts. In addition, a few showed the morphology of cell death, which seemed to be one of the reasons leading to the decrease of osteoclasts. There was no substantial change in the morphological relationships or ultrastructure of osteoclast precursor cells, stromal cells, extracellular matrices, and tumor cells between the experimental and the control groups. In the experimental group, the distribution of extracellular matrices (heparan sulfate proteoglycan and fibronectin) was less conspicuous, but the localization of osteotropic cytokines (interleukin-6 and prostaglandin E2) was essentially similar to that of the control group. The cause leading to the decrease of osteoclast precursor cells remains to be clarified. In conclusion, YM175 inhibits bone resorption induced by tumor, by decreasing the activity of mature osteoclasts and possibly affecting the production of osteoclast precursor cells. PMID- 8717530 TI - Bisphosphonates act on osteoblastic cells and inhibit osteoclast formation in mouse marrow cultures. AB - We examined the mode of action of bisphosphonates on osteoclastic cell recruitment using mouse marrow cultures with or without osteoblastic cells. Tartrate-resistant acid phosphatase-positive multinucleated cells [TRAP(+)MNC] formed in cultures were determined to be osteoclastic cells. In marrow cultures, TRAP(+) MNC formation in the presence of 10(-8) mol/L 1,25(OH)2D3 was not affected by the addition of 10(-6) mol/L dihydrogen (cycloheptylamino) methylenebisphosphonate monohydrate (YM175). However, it was inhibited in cocultures of marrow cells with osteoblastic cells. The inhibitory effect was evident throughout the entire culture period. YM175 dose dependently inhibited TRAP(+) MNC formation, and other bisphosphonates--pamidronate and alendronate- also inhibited TRAP(+) MNC formation in the coculture. Similar observations were also made in the coculture of spleen cells with osteoblastic cells. The conditioned media of osteoblastic cells treated with 10(-6) mol/L YM175 inhibited TRAP(+) MNC formation in marrow cultures. The presence of YM175 in methylcellulose cultures affected neither the colony formation of monocyte macrophage lineage, nor TRAP(+) MNC formation in the succeeding cocultures of recovered cells with osteoblastic cells. These results indicate that YM175 and probably other bisphosphonates as well preferentially inhibit the later stage of osteoclastogenesis through its action on osteoblastic cells. Our findings suggest that part of the inhibitory action by osteoblastic cells in the presence of bisphosphonates is mediated through soluble factor(s). PMID- 8717531 TI - No diffuse osteoporosis in lumbar scoliosis but lower femoral bone density on the convexity. AB - The aims of our study were to find out, in 15 female volunteers with untreated structural lumbar idiopathic scoliosis, through dual-photon X-ray absorptiometry measurements, whether femoral bone mineral density (BMD) was different between the right and the left side and whether this difference was related to scoliosis convexity. We showed that no statistical significant difference between femoral neck BMD measured on both sides of the same patient (p = 0.6). However, neck BMD from the same side as the convexity side was significantly lower than the opposite one. PMID- 8717532 TI - Changes of bone mineral mass and soft tissue composition after hip fracture. AB - The aim of this prospective longitudinal study was to measure prospectively the bone mineral density (BMD) and anthropometric variables after a hip fracture. In particular, we studied changes in the BMD in both the injured and uninjured hips, and examined if the postoperative mortality rate and complications, including pseudarthrosis of the fracture and late segmental collapse of the head of the femur, could be predicted by early bone mass measurements. The bone mineral density and the body composition were measured with dual energy X-ray absorptiometry in 102 consecutive hip fracture patients, 31 men and 71 women, with a mean age of 74 and 79 years, respectively. All cases were operated on within 3 days. The measurements were undertaken within 10 days after the fracture, after 4 and after 12 months. The BMD of the hip fracture cases decreased, especially in the lower extremities where the patients lost 7%, during the first year after the fracture. The patients also lost lean body mass (5%) but gained fat (11%) during the same period. They lost significantly more bone mass in the fractured hip than in the uninjured hip (p < 0.05). No difference was found between those patients who survived and those who died within 2 years after their hip fracture in neither the initial measurement nor in the follow-up measurements. Also, we found no difference between those patients whose hip fracture healed and those who developed late segmental collapse or pseudarthrosis. In conclusion, osteoporotic hip fracture cases lose bone mass at an increased rate, especially in the fractured hip. Also, their soft tissue composition changes, gaining fat while losing muscle mass. Furthermore, it seems that early bone mineral measurements cannot predict postoperative failures or postoperative mortality. PMID- 8717533 TI - Effects of altered diet on serum levels of 1,25-dihydroxyvitamin D and parathyroid hormone in X-linked hypophosphatemic (Hyp and Gy) mice. AB - X-linked hypophosphatemia is a metabolic bone disease occurring in both humans and mice. In mice, two different mutations (Hyp and Gy), occurring at separate but closely linked loci, have been proposed as models for this disease. Varying reports of the Vitamin D status of these two mutants has led us to reexamine the influence of diet on circulating calcitrophic hormones and mineral metabolism in both mutants. Hyp and Gy mice were raised on the B6C3H background, and both normal females and heterozygous mutant females were studied at 10 weeks of age. Animals were fed one of three diets at random: high (1.5% Ca and 1.0% P); medium (0.6% Ca and 0.6% P); or low (0.0% Ca and 0.6% P). After 3 days, serum and urine samples were collected. In comparison to mutant mice fed the high diet, both Hyp and Gy mice fed the low diet had decreased serum calcium levels, and further elevations in both serum alkaline phosphatase and serum parathyroid hormone (PTH). Serum 1,25-dihydroxyvitamin D levels were elevated by both the medium and low diets in all groups of mice over values obtained with the high diet. Mutant mice were significantly higher in serum PTH on all diets compared to normal mice fed the same diet. Mutant mice were not elevated in serum 1,25-dihydroxyvitamin D over normal mice when fed the high diet. However, both Hyp and Gy mice fed the medium and low diets were elevated in serum 1,25-dihydroxyvitamin D over normal mice. Serum PTH levels were correlated to serum 1,25-dihydroxyvitamin D levels with Hyp and Gy mice lying on the same line (r = 0.86; p < 0.0001). In summary, when Hyp and Gy mice are studied on the same genetic background and fed the same diet, similar responses are seen in PTH levels and 1,25-dihydroxyvitamin D levels. Both mutants should be useful in elucidating the pathophysiology of this poorly understood human disease. PMID- 8717534 TI - Immunohistochemical and ultrastructural localization of CGRP-positive nerve fibers at the epiphyseal trabecules facing the growth plate of rat femurs. AB - We performed immunohistochemical and ultrastructural studies to disclose a possible relationship between nerve fibers and bone metabolism. Immunohistochemical distribution of calcitonin gene-related peptide (CGRP) positive nerve fibers during bone development was assessed in the femurs of rats. CGRP-positive nerve fibers were denser in the epiphysis than in the metaphysis. These nerve fibers particularly ran along the epiphyseal trabecules facing the growth plate and came in contact with osteoclasts. Many osteoclasts at the epiphyseal trabecules facing the growth plate contained abundant toluidine blue and periodic acid-Schiff-positive granules. Electron microscopy revealed that these osteoclasts have many membrane-bound, electron-dense granular structures and dilated cisterns of rough endoplasmic reticulum containing electron-dense material. They were often surrounded by clear cells displaying features of nerve fiber and had no ruffled border. Furthermore, ultrastructural observations revealed electron-dense structures coating the cytoplasmic side of plasma membranes of the nerve fibers. We also observed coated pits in the cytoplasm of the osteoclasts facing the nerve fibers. To further clarify the role of innervation, we compared trabecules of rats undergoing denervation of the sciatic nerve with those from unoperated rats. Denervation resulted in a significant increase in the number of cement lines on the epiphyseal trabecules facing the growth plate. These results suggest that the osteoclasts at the epiphyseal trabecules facing the growth plate are in part regulated by CGRP-positive nerve fibers. Thus, CGRP-positive nerve fibers could be a crucial element in bone metabolism during bone growth and development. PMID- 8717535 TI - Determinants of hip axis length in women aged 10-89 years: a twin study. AB - Hip axis length (HAL), a measure of femoral geometry, has been shown to predict hip fracture in white women over the age of 67 years, independently of bone mineral density at the femoral neck. A cross-sectional study of 304 pairs of female twins [176 monozygous (MZ) and 128 dizygous (DZ)], aged between 10 and 89 years, was performed to examine the influence of age, constitutional, lifestyle, and genetic factors on HAL. HAL was calculated from dual energy X-ray absorptiometry scans of the proximal femur using an automated technique with an Hologic QDR-1000W. Lean mass, fat mass, height, and weight were also measured. Maximum mean HAL was achieved by the age of 15 years. After this age there was no discernible dependency of mean HAL on age. Using within-pair differences, after adjusting for height there were no other independent constitutional or lifestyle predictors. Cross-sectionally, after adjustment for height, MZ and DZ correlations were 0.79 (95% CI: 0.73-0.84) and 0.54 (95% CI: 0.39-0.68), respectively, and independent of age. The MZ correlation exceeded the DZ correlation (p < 0.001). The best-fitting model apportioned 79% (SE 7%) of variation in height-adjusted HAL to additive genetic factors. There was marginal evidence that an environmental influence shared by twins explained 31% (SE 16%) of height-adjusted variance (p = 0.07), in which case the genetic variance was reduced to 51% (SE 15%). Adjustment for height had reduced the magnitude of total variance by 26%, and 95% of this reduction was in the additive genetic component. Applying a previously described theoretical model, approximately 10% of the increased risk of hip fracture associated with a maternal history of hip fracture could be attributed to the genetic factors determining HAL. We conclude that, in women, adult HAL is achieved by midadolescence. After adjustment for height, which is itself largely under genetic influence, other genetic factors appear to play the predominant role in explaining variation in HAL. PMID- 8717536 TI - Erythroid-specific expression of human growth hormone affects bone morphology in transgenic mice. AB - The regulation of bone deposition and remodeling is highly complex. To further understand the influence of growth hormone on bone deposition, several lines of transgenic mice were generated that expressed the human growth hormone gene (hGH) driven by beta-globin regulatory elements. In situ hybridization confirmed that the hGH gene in these mice was expressed in an erythroid tissue-specific manner; in the fetus hGH was expressed in the liver and in the adult mice hGH was expressed in the bone marrow. The bones of mice in two lines were visualized radiographically by mammography, and relative bone densities were measured. The transgenic mice had detectably more bone density than nontransgenic littermate controls by approximately 3 weeks of age and the relative difference in density increased with age. Histological cross-sections of the tibia showed that adult transgenic mice had increased average cortical bone thickness when compared to their controls. The hypothesis is that the local effect of hGH release from differentiating erythroid cells in the bone marrow is a major contributor to the increased bone deposition in these transgenic mice. PMID- 8717537 TI - Bone morphogenetic protein-2 but not bone morphogenetic protein-4 and -6 stimulates chemotactic migration of human osteoblasts, human marrow osteoblasts, and U2-OS cells. AB - Bone morphogenetic proteins (BMPs) have important functions for the differentiation of bone cells, but the exact role for bone remodeling and bone healing still needs to be defined. Migration of bone forming cells is an important physiological event both during bone healing and bone remodeling. The chemotatic properties of the bone morphogenetic protein family of growth factors have not been investigated. In this study the chemotactic effects of the bone morphogenetic proteins BMP-2, -4, and -6 have been quantitated toward human osteoblasts, human marrow stromal osteoblasts, and U2-OS human osteosarcoma cells. BMP-2 stimulated the migration of human stromal osteoblasts, human osteoblasts, and U2-OS cells with bell-shaped response curves in a dose-dependent manner with a 300% increase in cell migration at 1.0 ng/mL for human stromal osteoblasts and a 170-180% increase for human osteoblasts and U2-OS cells. At higher concentrations, migration decreased to background levels. BMP-4 and -6 did not show any effect on cellular migration. This study shows that BMP-2 can stimulate in vitro migration of human osteoblasts and human osteosarcoma cells. BMP-2 might play a role in the chemotactic recruitment of especially undifferentiated osteoblasts during bone remodeling and bone healing. PMID- 8717538 TI - Effects of parathyroid hormone and agonists of the adenylyl cyclase and protein kinase C pathways on bone cell proliferation. AB - The anabolic effect of parathyroid hormone (PTH) on bone is partly due to a stimulation of osteoblast proliferation. The PTH signal is transduced by the pathways of adenylyl cyclase (AC)/protein kinase (PK) A and phospholipase C/PKC/Ca++. There is still uncertainty about the relative contribution of the two pathways to the proliferative effects of the hormone. In our study, PTH(1-34), AC/PKA agonists, and phorbol 12-myristate-13-acetate (PMA, a PKC activator) stimulated cell proliferation in cultured mouse calvariae. In isolated osteoblasts, only PMA stimulated proliferation, whereas AC/PKA agonists and PTH(1 34) inhibited it. As already known, PTH in the presence of supramaximal concentrations of transforming growth factor-beta (TGF-beta) stimulated osteoblast growth; under these same conditions, AC/PKA agonists reproduced the stimulatory effect of PTH(1-34), whereas PMA became inhibitory. PTH(1-31), which stimulates AC without affecting PKC, acted similarly to the fully active PTH(1 34) in both calvaria and isolated osteoblasts. On the contrary, midregion fragments that activate only PKC stimulated calvaria cell proliferation faintly in comparison with PTH(1-34); no effect was seen in osteoblasts, either with or without TGF-beta. Our study shows that the effects of PTH on proliferation can be mimicked by agonists of the AC/cAMP pathway. Although PMA is indeed able to stimulate cell growth in tissue explants, its effects on isolated osteoblasts markedly diverge from those of PTH. We conclude that activation of the AC/PKA pathway is the main component of the proliferative effects of PTH. PMID- 8717539 TI - Cortical bone remodeling in autosomal dominant osteopetrosis: a study of two different phenotypes. AB - Previous analyses of cancellous bone remodeling in autosomal dominant osteopetrosis (ADO) have suggested a diminished osteoclastic resorption at the endocortical surface. Consequently, cortical width increases with age in both ADO type I and II. By means of histomorphometric methods, the remodeling cycle of the Haversian surface in the iliac crest has recently been reconstructed in normal individuals. The aim of the present investigation was to study cortical bone remodeling in ADO type I and II. Transcortical iliac crest bone biopsies obtained from 21 ADO patients were studied. Of these, 14 had ADO type I and seven had ADO type II. Twenty of the patients had received intravital double labeling with tetracycline before the biopsies were obtained. The static histomorphometric parameters for cortical bone mass and structure were compared with a control group of 21 sex- and age-matched individuals. Regarding the dynamic histomorphometric parameters for reconstruction of the cortical bone remodeling cycle, the ADO type I material was compared with control material from 14 sex- and age-matched individuals, whereas the ADO type II material was compared with control material from six sex-matched but younger individuals. Significant increases were seen in absolute as well as fractional cortical widths in both ADO types. Furthermore, an age-related increase was observed in both absolute and fractional cortical widths in ADO type I, whereas ADO type II was nonsignificant with regard to age. The fractional cancellous width was reduced in both type I and type II. However, only in ADO type I did the fractional cancellous width significantly correlate inversely with age. In the control group, neither cortical dimensions nor cancellous width correlated significantly with age. No significant differences were observed between patients and the control group in osteon dimensions; fractional resorptive, formative, or quiescent sites; resorptive or formative remodeling rates; remodeling periods; or activation frequency. An age-related increase in cortical porosity was seen in the group of normal individuals, but not in the two patient groups. The fractional remodeling space was increased in the ADO type II group. In conclusion, cortical dimensions are increased in both ADO type I and II and positively correlated to age in type I. However, cortical remodeling at the Haversian surface is essentially normal in both ADO type I and II. This could be explained by a defective endocortical bone resorption, as no periosteal accretion was observed, and because cancellous bone remodeling previously has been described to be normal. PMID- 8717540 TI - Physical loading, exercise, and bone. PMID- 8717541 TI - Mechanical factors in bone growth and development. AB - Mechanobiologic factors strongly influence skeletal ossification and regulate changes in bone geometry and apparent density during ontogeny. We have developed computer models that implement a simple mathematical rule relating cyclic tissue stresses to bone apposition and resorption. Beginning at the fetal stages of the femoral anlage, these models successfully predict the appositional bone growth and modeling observed in the development of the diaphyseal cross section. The same basic mechanobiologic rule can also predict the architectural construction of the proximal cancellous bone formed in regions of endochondral ossification. Geometry and density changes in adult diaphyseal and cancellous bone as a result of changes in physical activity can be simulated by invoking the same rule used during development. Future clinical and experimental work is needed to provide more quantitative data for mechanobiologic rules and elucidate the interactions between chemical and mechanical factors influencing bone biology. PMID- 8717542 TI - Endogenous and nutritional factors affecting bone. PMID- 8717543 TI - Biochemical assessment of bone resorption and formation. PMID- 8717544 TI - Estimation of various mechanical characteristics of human bones using dual energy X-ray absorptiometry: methodology and precision. AB - The mechanical competence of bone is determined by its macroscopic geometric characteristics [size, shape, apparent density (BMAD), cortical thickness (CWT), cross-sectional area, trabecular architecture], intrinsic material properties (stiffness, strength), and loading conditions (mode, direction, rate) at a given skeletal site. The main objective of this study was to introduce precise methods for further analysis of dual energy X-ray absorptiometric (DXA) data and for estimating macroscopic mechanical characteristics of bone at several skeletal sites representing both the axial and appendicular skeleton. This study showed that using computational BMAD, CWT and dimensional parameters, different site specific mechanical characteristics (stiffness and strength indices) of a typical long bone (a bone consisting of both the cortical shaft and trabecular metaphyses and epiphyses at both ends) can be approximated with a low precision error (generally < 2%). The efficacy of a study applying DXA may be further enhanced by applying relevant site-specific parameters (different parameters for bone ends and shafts) for evaluation, the parameters based on the anatomic ROIs. Evidently high-quality operator performance is a prerequisite for effective accomplishment of longitudinal studies, especially when small changes in bone characteristics are expected. It should be kept in mind that to some extent the inherent two dimensional nature of the DXA measurement compromizes the potential efficacy of the presented methods since some simple assumptions had to be made regarding the geometry and structure of bone. Nevertheless, in this study the observed values were consistent with those found by other investigators. Further experimental studies are needed for direct site-specific validation of the proposed analytic methods. PMID- 8717545 TI - In vivo assessment of bone mechanical properties by vibration and ultrasonic wave propagation analysis. AB - Vibration analysis and ultrasonic wave propagation analysis were evaluated as noninvasive techniques for the in vivo assessment of bone mechanical properties. The relation between the resonant frequencies, obtained by vibration analysis, and geometrical and material properties of long bones is explained using a simple beam model. This simple beam model was validated experimentally in previous work on excised animal bones. In vitro measurements were performed on human and animal excised bones from specific osteopenic cases and control groups. Using specific protocols for in vivo vibration and ultrasound measurements of the tibia, a population of osteoporotic patients and age-matched controls were tested. From these measurements, it was concluded that the bending rigidity, calculated from the resonant frequencies, in osteoporotic tibiae had decreased as compared to the control group. Also the ultrasound velocity in the tibial cortex was lower in the osteoporotic group. The latter indicates a change in the bone tissue material properties. On the other hand, immobilization osteoporosis appeared to lead to a decrease in bending rigidity without an observable change in bone tissue material properties. By the combination of vibration analysis and ultrasound velocity measurements, the whole bone's mechanical characteristics as well as the bone tissue properties can be assessed in vivo. Since both techniques are noninvasive, they can be used in longitudinal studies for the assessment of bone response on physical loading. PMID- 8717546 TI - Using functional loading to influence bone mass and architecture: objectives, mechanisms, and relationship with estrogen of the mechanically adaptive process in bone. AB - There is increasing evidence that load-bearing is an important, if not the most important, functional influence on bone mass and architecture. Load-bearing most probably exerts its influence through the dynamic strains engendered in the bone tissue. Mechanically adaptive bone modeling and remodeling can be regarded as a homeostatic mechanism regulating functional bone strains at each location throughout the skeleton. Because most long bones are loaded in a certain amount of bending normal function, strains vary across the bones' cross-section. Both the longitudinal curvature and cross-sectional shape of a number of bones engender strains during functional loading rather than reduce them. Bone's adaptive response to load-bearing therefore results in functional strains which are neither uniform in distribution nor minimal in magnitude. Not all aspects of bone's strain environment are equally effective as influences on bone architecture. Unusual strain distributions, high strains, and high strain rates seem to be particularly osteogenic. The osteogenic response which follows exposure to such strains appears to saturate after only a few loading cycles. This is consistent with adaptive bone (re)modeling being sensitive to strain "errors," which are not repeated frequently, rather than the repetitious strain cycles engendered during normal predominant activities. Exercise regimens designed to control bone architecture can usefully capitalize on this feature of the adaptive (re)modeling response. Each exercise session need not be prolonged but should include as many novel strain distributions as possible, preferably involving high peak strains and strain rates. To maintain any level of bone mass requires a continued, loading-related osteoregulatory stimulus. Exposure to appropriate load-bearing exercise needs therefore to be repeated, probably at daily or alternate-daily intervals. In short-term experiments in rat bones, estrogen amplifies the osteogenic response to a single period of loading. The features of postmenopausal bone loss are consistent with the etiology of the condition being primarily withdrawal of estrogen's contribution to bone's mechanically adaptive response. PMID- 8717547 TI - Dose-response relationship between physical loading and mechanical competence of bone. AB - A dose-response relationship between mechanical loading and bone can be inferred by the fact that bone response is proportional to the applied load in cell and organ culture and in animal models where a known load at a given magnitude and frequency is applied. Proportional responses have been observed in second messengers, growth factors, bone matrix, and bone strength. In the human model, however, the determination of a dose-response relationship is hindered by the lack of appropriate technology to directly evaluate the mechanical load and the skeletal competence. In vitro loading of bone does not duplicate the in vivo physiologic conditions, in particular the neuromuscular responses to the loads applied to the skeleton. While animal studies aid in determining the mechanisms of bone response to mechanical loading and dietary interactions, they do not obviate the need for human clinical trials to evaluate the effects of physical activity on both skeletal competence and fall prevention. The literature reviewed shows that physical activity increases the competence of the skeleton to resist fracture by the maintenance and improvement of bone mineral density (BMD) and neuromuscular competency, thus reducing skeletal fragility, pre-disposition to falls, and fall impact. PMID- 8717548 TI - Exercise and bone mass in young and premenopausal women. AB - The studies that investigate exercise effects on bone mass in young and premenopausal women are sparse. This population of women represents three distinct subgroups: the young woman just at the end of longitudinal growth, the adult woman who is beginning to lose bone, and the competitive female athlete. The majority of research has focused on the female athlete and most investigators report lower than normal bone mass among women who experience exercise-associated oligo- and amenorrhea. More recent investigations have reported that the type of exercise has a profound effect on bone mineral density and activities that deliver high loads appear to offset the effects of low reproductive hormones. Information gleaned from these studies has important implications for the design of exercise programs to build bone mass in the young adult woman and offset bone loss in the older premenopausal woman. PMID- 8717549 TI - Epidemiology of hip fractures. AB - There were an estimated 1.66 million hip fractures world-wide in 1990. According to the epidemiologic projections, this worldwide annual number will rise to 6.26 million by the year 2050. This rise will be in great part due to the huge increase in the elderly population of the world. However, the age-specific incidence rates of hip fractures have also increased during the recent decades and in many countries this rise has not leveled off. In the districts where this increase has either showed or leveled off, the change seems to especially concern women's cervical fractures. In men, the increase has continued unabated almost everywhere. Reasons for the age-specific increase are not known: increase in the age-adjusted incidence of falls of the elderly individuals with accompanying deterioration in the age-adjusted bone quality (strength, mineral density) may partially explain the phenomenon. The growth of the elderly population will be more marked in Asia, Latin America, the Middle East, and Africa than in Europe and North America, and it is in the former regions that the greatest increments in hip fracture are projected so that these regions will account for over 70% of the 6.26 million hip fractures in the year 2050. The incidence rates of hip fractures vary considerably from population to population and race to race but increase exponentially with age in every group. Highest incidences have been described in the whites of Northern Europe (Scandinavia) and North America. In Finland, for example, the 1991 incidence of hip fractures was 1.1% for women and 0.7% for men over 70 years of age. Among elderly nursing home residents, the figures can be as high as 6.2% and 4.9%. The lifetime risk of a hip fracture is 16%-18% in white women and 5%-6% in white men. At the age of 80 years, every fifth woman and at the age of 90 years almost every second woman has suffered a hip fracture. Since populations are aging worldwide, the mean age of the hip fracture patients are increasing rapidly, too. Between 1970 and 1991, the mean age of male Finnish patients increased dramatically from 52.9 years to 69.0 years. In women, the corresponding figures were 71.6 and 78.9 years. This change is likely to cause increasing problems in the treatment and rehabilitation of the patients. In 1990, 72% of the hip fractures worldwide occurred in women. All over the world, the hip fracture incidences are about two times higher in women than in men. Women's overrepresentation has been explained by women's lower bone mass and density and higher frequency of falling. Epidemiologic studies show that trochanteric fractures are an increasing problem since compared with cervical fractures their relative number increases progressively with age in women after the age of 60 years and since their incidence has been shown to increase in both sexes and all age groups during the recent decades. This may have direct public health implication since mortality, morbidity, and costs caused by trochanteric fractures are higher than those of the cervical fractures. Reduced bone density (strength) by age and over the recent decades has been the most frequently mentioned reason for the increase of trochanteric fractures. Also, the fall characteristics of the elderly may have changed during the recent decades resulting in increasing numbers of this type of hip fractures since the type of the hip fracture (cervical or trochanteric) also depends on the impact angle of the greater trochanter at the moment of the floor contact. PMID- 8717550 TI - Hip fractures: incidence, risk factors, energy absorption, and prevention. AB - The present review summarizes the pathogenic mechanisms leading to hip fracture based on epidemiological, experimental, and controlled clinical studies. The estimated lifetime risk of hip fracture is about 14% in postmenopausal women and 6% in men. The incidence of hip fractures increases exponentially with aging, but the time trend in increasing age-specific incidence may finally reach a plateau. Postmenopausal women suffering earlier non-hip fractures have an increased risk of later hip fracture. The relative risk is highest within the first years following the fracture. Nursing home residents have a high risk of hip fracture (annual rate of 5-6%), and their incidence of falls is about 1.5 falls/person per year. Most hip fractures are a result of a direct trauma against the hip. The incidence of falls on the hip among nursing home residents is about 0.29 falls/person per year and about 20% of these traumas lead to hip fracture. Women with hip fractures have a lower body weight compared with controls, and they may also have less soft tissue covering the hip, even when adjusted for body mass index, indicating a more android body habitus. Experimental studies show that the passive energy absorption in soft tissue covering the hip may influence the risk of hip fracture and be an important determinant for the development of hip fracture, perhaps even more important than bone strength. External hip protectors were developed and tested in an open randomized nursing home study. The rate of hip fracture was reduced by 50%, corresponding to 9 of 247 residents saved from sustaining a hip fracture. This review points to the essentials in the development of hip fracture: risk of fall; type of fall; type of impact; energy absorption; and last, bone strength, which is the final permissive factor leading to hip fracture. Risk estimation and prevention of hip fracture may prove realistic when these issues are taken into consideration. PMID- 8717551 TI - Etiology and prevention of age-related hip fractures. AB - Falls and fall-related injuries are among the most serious and common medical problems experienced by the elderly. Hip fracture, one of the most severe consequences of falling in the elderly, occurs in only about 1% of falls. Despite this, hip fracture accounts for a large share of the disability, death, and medical costs associated with falls. As measured by their frequency, influence on quality of life, and economic cost, hip fractures are a public health problem of crisis proportions. Without successful international initiatives aimed at reducing the incidence of falls and hip fractures, the implications for allocations of health resources in this and the next century are staggering. Identifying those at risk for harmful falls requires an understanding of what kinds of falls result in injury and fracture. In elderly persons who fall, in most of whom hip bone mineral density is already several standard deviations below peak values, fall severity (as reflected in falling to the side and impacting the hip) and body habitus are important risk factors for hip fracture and touch on a domain of risk entirely missed by knowledge of bone mineral density. These findings clearly suggest that factors related to both loading and bone fragility play important roles in the etiology of hip fracture. We provide a strategy, based on engineering approaches to fracture risk prediction, for determining the relative etiologic importance of loading and bone fragility and to summarize some of what is known about both sets of factors. We define a factor of risk, phi, as the ratio of the loads applied to the hip divided by the loads necessary to cause fracture and summarize available data on the numerator and the denominator of phi. We then provide an overview of the complex interplay between the risks associated with the initiation, descent, and impact phases of a fall, thereby suggesting an organized approach for evaluating intervention efforts being used to prevent hip fractures. The findings emphasize the continuing need for combined intervention strategies that focus on fall prevention, reductions in fall severity, and maintaining or increasing femoral bone mass and strength, either through targeted exercise programs, optimal nutrition (Ca, Vitamin D), and/or in the use of osteodynamic agents. By developing and refining the factor of risk, a property that captures both the contributions of bone density and the confounding influences of body habitus and fall severity, we believe these intervention strategies can be targeted more appropriately. PMID- 8717552 TI - Prevention of falls in the elderly. AB - The purpose of these symposium presentations was to examine the role of physical activity as a means of preventing hip fractures through the prevention of falls. Risk factor identification is necessary to develop preventive strategies. Risk factors related to physical activity and other risk factors for falls were identified. Intervention studies aimed at reducing, preventing or delaying falls were identified and discussed. A literature search from 1976-1994, identified 52 studies examining risk factors for falls, recurrent falls and/or falls resulting in injury. Nine intervention studies were identified with the primary outcome of falls. Physical activity-related risk factors for falls include limitations in general functioning, such as ambulation and mobility problems, difficulty or dependence in activities of daily living, and exposures to the risks of falling as indicated by the nature and frequency of daily activities. Impairments in gait and balance as well as neuromuscular and musculoskeletal impairments frequently underlie changes in physical activity in old age. Reduced activity level may occur as a result of these impairments, leading to further declines in physical functioning and an increased risk of falls. A relatively high level of activity in old age is also associated with risk of falls. Other risk factors for falls, such as cognitive impairment, visual deficits and medication use, may combine with physical activity-related risk factors to increase the risk of falls. Intervention studies directed at nursing home populations did not prevent falls but had other statistically and clinically significant outcomes. Studies among the community dwelling that targeted potential or current risk factors and included an exercise component reported a significant reduction in falls, prevented the onset of new disabilities and reduced baseline risk factors. Prevention of falls and subsequent injuries in the institutionalized population remains a challenge. Further development of interventions for community-dwelling elders that facilitate maintenance of physical activity without unduly increasing the risk of falls is also critical. The potential for maintenance of benefits gained from all fall interventions needs further examination. PMID- 8717553 TI - Prevention of preeclampsia with heparin and antiplatelet drugs in women with renal disease. AB - In a retrospective cohort study of women with renal disease in pregnancy we investigated if: 1. low dose aspirin reduced the prevalence of preeclampsia and improved fetal outcome compared to no anticoagulant therapy. 2. heparin plus low dose aspirin and/or dipyridamole reduced the prevalence of preeclampsia and improved fetal outcome compared to i. no treatment ii. low dose aspirin alone. Women with renal disease were allocated into 3 groups according to the treatment received during their pregnancies: I. no prophylactic heparin or antiplatelet drugs, n = 76 II. prophylactic low-dose aspirin 75(50-150)mg, n = 27 III. prophylactic subcutaneous heparin 10,000 (5000-12,500) IU b.d. combined with low dose aspirin 50 (50-150)mg and/or dipyridamole 400 (200-400)mg, n = 44. Preeclampsia and fetal outcome was analysed according to treatment group. Preeclampsia was less common in the heparin group (2.3%) compared with 27.6% in the no treatment group [O.R. 0.06 (0.01-0.30)] and 25.9% in the aspirin group [O.R. 0.07 (0.01-0.38)]. Women on aspirin, who developed preeclampsia, delivered later in pregnancy [35.4 (33-38.2) weeks] than preeclamptic women on no treatment [29 (22-38) weeks], p = 0.04. There was a trend to reduced perinatal deaths in the heparin + antiplatelet drug group, [2.3%; O.R., 0.17 (0.02-1.4)] and in the aspirin group [0%, O.R., 0.13 (0.01-2.3)] compared with 11.7% perinatal deaths in the no treatment group. Heparin with anti-platelet drugs may prevent preeclampsia in high risk women with renal disease. Further investigation in a randomized trial is indicated. PMID- 8717554 TI - Transient blindness associated with hypertensive disorders in pregnancy. PMID- 8717555 TI - Inadequacy of dipstick proteinuria in hypertensive pregnancy. AB - The objective of this study was to determine the accuracy of ward urinalysis and the sensitivity of dipstick testing in the assessment of proteinuria in hypertensive pregnant women. Subjects were 230 consecutive hypertensive pregnant women who were admitted to hospital over a 2-year period. Routine ward urinalyses for protein, obtained on a mid-stream sample before and after a 24-hour urine collection for quantitating proteinuria, were compared with the 24-hour urine protein excretion. As a control for dipstick accuracy, urinalysis was also performed on a mixed aliquot of each of the 24-hour samples by a single observer experienced in urinalysis. True proteinuria was considered as > 300 mg/day. The positive predictive value for urinalysis ranged from 38% (for the precollection test) to 60% (for tests on the aliquot). Negative predictive values were 86-88%. The false negative rates at 'nil' or 'trace' proteinuria ranged from 8-18%. The false positive rates at '3+' (3 g/L) or '4+' (> or = 20 g/L) ranged from 0-17%, at '2+' (1 g/L) from 18-50% and at '1+' (0.3 g/L) from 67-83%. Best results for urinalysis were obtained on the aliquot testing but even under these ideal circumstances there was a high false positive rate (67%) at '1+' (0.3 g/L) urinalysis level. These studies show that in routine clinical practice 'nil' or 'trace' proteinuria will miss significant proteinuria in approximately 1 out of 8 hypertensive pregnant women while '3+' (3 g/L) or '4+' ( > or = 20 g/L) will rarely be a false positive. At urinalysis of '1+' or '2+' a 24-hour urine collection is required to be certain about the presence or absence of proteinuria. Research studies should demand 24-hour urine protein quantitation and not rely solely upon urinalysis results. PMID- 8717556 TI - Low dose aspirin for the treatment of fetal growth restriction: a randomized controlled trial. AB - The purpose of this study was to investigate the hypothesis that maternal administration of 100mg aspirin each day will improve birth-weight and other measures of neonatal size when given as a treatment to pregnancies complicated by fetal growth restriction and umbilical-placental insufficiency. A randomized, double-blind, placebo controlled study design was employed; 51 pregnant women were enrolled. The entry criteria were a fetal abdominal circumference < 10th per centile together with an umbilical artery Doppler systolic/diastolic ratio > 95th per centile between 28 and 36 weeks' gestation. Compliance was assessed by serial measurement of maternal serum thromboxane B2 levels. The mean gestational age at enrolment was 32 weeks and at delivery was 36 weeks. There were no differences between the 2 groups in gestational age at birth; birth-weight or birth-weight ratio; circumferences of the head, chest or abdomen; skin fold thicknesses; or neonatal morbidity. Low dose aspirin therapy did not alter Doppler systolic/diastolic ratios. After 14 days therapy, mean thromboxane B2 levels fell more than 80% from baseline values; 10.5% of women did not demonstrate biochemical confirmation of aspirin ingestion, despite verbal confirmation of compliance. We conclude that low dose aspirin therapy is not of benefit in the treatment of pregnancies complicated by fetal growth restriction and umbilical placental insufficiency between 28 and 36 weeks' gestation. PMID- 8717557 TI - The prevalence and timing of obstetric ultrasound in Victoria 1991-1992: a population-based study. AB - A statewide study to ascertain the number of ultrasound scans received by women in pregnancy, to identify the proportion having a scan at 16 to 20 weeks' gestation, and to establish where the scan at 16 to 20 weeks was performed was carried out between January, 1991 and June, 1992 in Victoria. Additional data were collected by midwives and entered on the perinatal morbidity statistics form routinely completed for all births. Of 52,319 women providing responses, 3.1% did not have a scan. Of the remaining 96.9% who had a scan, 73.5% were scanned at 16 to 20 weeks' gestation. Predictors of not having a scan were maternal birthplace and higher parity: previous perinatal death(s), and attendance at nonteaching hospitals predicted the opposite. Predictors of being scanned were location of hospital (country), maternal birthplace, higher parity and maternal age (< 20 years). Substantial differences in frequency and timing were found between hospitals attended. Factors associated with the pattern of scanning are not readily explicable in terms of risk of malformations or women's choices. PMID- 8717558 TI - The effect of work activity in pregnancy on the risk of severe preeclampsia. AB - The aim of this study was to evaluate the impact of type of employment and level of physical activity at work on the risk of severe preeclampsia. For this purpose, we carried out a case control study of 160 nulliparous pregnant women with severe preeclampsia and 320 normotensive nulliparous controls who received prenatal care from members of one medical staff. The type of employment and the level of physical activity sustained at work were ascertained at birth through an interview based on a standard questionnaire. The degree of physical activity at work was assessed by a 4-level activity score based on type of work, physical intensity, posture at work, and weekly working hours. In logistic regression analysis, after adjustment for potential confounders (maternal age, time of stopping work, prepregnancy body mass index, social status of the partner, history of previous abortion, and prepregnancy smoking status), clerical workers had a significantly lower risk of severe preeclampsia than women who were unemployed at the beginning of pregnancy (OR 0.53, 95% CI 0.30, 0.96). In multivariate analysis, there was a significant linear trend relating the degree of physical activity at work, to the risk of preeclampsia (likelihood chi-square = 9.38, 3 df, p = 0.002). We then restricted the analysis to women who had ever worked in pregnancy (n = 339) also adjusting for confounders, and found that clerical workers were still at significantly lower risk of severe preeclampsia than women not formally employed (OR 0.2, 95% CI 0.08, 0.49). In addition, moderate/high physical activity at work was associated with a 2-fold increase in the risk of severe preeclampsia compared to mild activity (OR 2.08, 95% CI 1.11, 3.88). We conclude that moderate to high physical activity at work seems to increase the risk of severe preeclampsia. PMID- 8717560 TI - Gestational diabetes: do all women need to be tested? AB - The results of glucose tolerance testing in 1,185 consecutive women were examined with respect to historical risk factors for gestational diabetes mellitus (GDM). GDM was present in 79 of 1,185 (6.7%) of the women. GDM was present in 8.5% of women aged > or = 30 years, in 12.3% of women with a preconception BMI > or = 30 and in 11.6% of women with a family history of diabetes in a first degree relation. A combination of one or all of these risk factors predicted GDM in only 48 of 79 (60.8%) cases. GDM was present in 4.8% of women without risk factors. Testing for GDM on the basis of these historical risk factors alone, and using the Australasian Diabetes in Pregnancy Society (ADIPS) criteria for diagnosis, would miss more than one-third of all cases. This study supports the ADIPS recommendation that there should be universal testing. PMID- 8717559 TI - Anti-D administration in early pregnancy - time for a new protocol. AB - The use of anti-D immunoglobulin has dramatically reduced the incidence of rhesus isoimmunization arising from pregnancy. A critical shortage of anti-D has developed and consumption must be reduced, preferably without increasing the risk of isoimmunization. The standard dosage of anti-D currently given at all gestational ages is 1 ampoule containing 125 micrograms of anti-D. Of the anti-D administered half is during the first trimester when a much lower dose could be used; 30 micrograms would remove from the maternal circulation the entire fetoplacental blood volume (3 ml) of a 12 week's pregnancy. A dose of 30 micrograms would also be adequate for most fetomaternal transfusions after 12 weeks but prior to the third trimester. A larger dose could be administered if maternal Kleihauer testing suggests the need for a supplementary dose. In the third trimester, the current regimen is appropriate - a dose of 125 micrograms of anti-D supplemented when needed on the basis of maternal Kleihauer testing. A lower dose ampoule of anti-D should be available. In addition, existing protocols for anti-D administration should be modified so that smaller more appropriate doses are given, better utilizing the limited supply of anti-D. PMID- 8717561 TI - The effects of antenatal steroid use in premature rupture of membranes. AB - We sought to determine if antepartum steroid treatment offers any clinical benefits to patients with premature rupture of membranes. One hundred and forty five maternal-neonatal pairs were studied. Forty-five maternal-neonatal pairs with premature, preterm rupture of membranes received steroids during 24-35 weeks' gestation. One hundred maternal-neonatal pairs received no antenatal steroids. The 2 groups were identical with regard to gestational age at rupture of membranes, gravity, parity, race, fetal gender, socioeconomic status, smoking, and preterm labour risk factors. Study of the data revealed that maternal chorioamnionitis was less frequent in the steroid group (p < 0.001). Bronchopulmonary dysplasia (oxygen dependent at discharge at term gestational age) was less frequent in the steroid group (p < 0.05). The remainder of the data revealed no statistically significant differences in preterm delivery rate, necrotizing enterocolitis, respiratory distress syndrome, intraventricular haemorrhage rate or severity of haemorrhage, hospital days, latency to delivery, or ventilator days. Antepartum steroid use in preterm rupture of membranes appears to offer clinical benefit in premature infants by lessening the rate of bronchopulmonary dysplasia in those infants receiving antepartum steroids. PMID- 8717562 TI - A study of breast feeding rates at a large Australian obstetric hospital. AB - A prospective breast feeding survey in a large obstetric hospital was carried out from 1988 to 1991. For each year, a sample of women were interviewed following delivery and data was collected which included the method of feeding, patient status (public or private patients), age, parity, (including previous breast feeding experience), marital status, country of birth and the number of babies. The mother's feeding method after delivery and on discharge from hospital were recorded. Women who were breast feeding on discharge were interviewed at 3 months. When putting the figures for the 4 years together, the breast feeding commencement rate was 88%, the breast feeding rate on discharge was 80% and the breast feeding rate at 3 months was between 51% and 57%. Factors found to be affecting the breast feeding rate at 3 months included patient status, age and parity. Problems experienced by the mothers after discharge from hospital included nipple pain, nipple trauma and mastitis. Private patients reported a significantly higher rate of mastitis than public patients. PMID- 8717563 TI - A pain in the neck in pregnancy: cervical spinal tuberculosis. AB - This paper describes a case of spinal tuberculosis diagnosed at 29 weeks' gestation in a primigravid Indian woman. The report illustrates the primary problems in suspecting disease, recognizing the diverse clinical presentations and initiating investigations to confirm the diagnosis. In most cases, M. tuberculosis, once recognized and appropriately treated, should result in an excellent prognosis. PMID- 8717564 TI - Management guidelines for M. tuberculosis in pregnancy. AB - Unrecognized M. tuberculosis in pregnancy can have an adverse outcome for women and their babies. With world-wide trends of increasing incidence of M. tuberculosis and increasing migration to Australia of women of childbearing age from countries with a high incidence of this disease, it is imperative that clinicians recognize and be aware of appropriate management of the disease. Management guidelines for M. tuberculosis occurring in pregnancy are outlined. PMID- 8717565 TI - Indications for blood transfusion in pregnancy with sickle cell disease. AB - In order to assess the value of prophylactic blood transfusion in the management of pregnant patients with sickle cell disease, a study was conducted on 571 sickle cell disease pregnancies. These patients were divided into 2 groups, one with prophylactic transfusion and the other with restricted transfusion. In spite of the reduction in the incidence of blood transfusion the rate of preterm labour, intrauterine growth retardation, perinatal mortality and maternal mortality were similar in both groups. A restricted blood transfusion policy can be safely followed without compromising fetal or maternal well-being with an additional benefit of reducing the incidence of transfusion related crises and other complications. PMID- 8717566 TI - Invasive cervical cancer in pregnancy. AB - Cervical cancer is the commonest malignancy which complicates pregnancy, but the management remains controversial. We reviewed our patients in an attempt to identify the best management options which resulted in long-term survival for the mother and a live baby. The total number of pregnancies managed between January, 1981 and March, 1995 was obtained from the hospital records, and patients with invasive cervical cancer diagnosed during pregnancy or within 12 months of delivery were identified. The case records were reviewed. Between January, 1981 and March, 1995 there were 22 cases of cervical cancer diagnosed either during pregnancy or within 12 months postpartum. This gave an incidence of cervical cancer associated with pregnancy of 1 in 3,817 pregnancies or 0.26 per 1,000 pregnancies. Eleven patients had microinvasive disease. Nine were treated by cone biopsy and 2 by radical hysterectomy. Nine patients had Stage 1B and 1 had Stage 2A disease and all were treated with radical hysterectomy. One patient had Stage 3B disease and was treated with radiotherapy and chemotherapy followed by simple hysterectomy. Fourteen patients delivered vaginally. Twenty of the 22 patients were delivered of live babies which survived. The patients have been followed from 1 month to 13 years with only 1 recurrence, and all 22 remain alive. We conclude that all pregnant women should have a Pap smear performed antenatally. Cone biopsy can be safely performed in pregnancy and may be adequate treatment for microinvasive squamous cell carcinomas. Treatment, including the timing of delivery, must be individualized, with the patient playing an important decision making role. PMID- 8717567 TI - A randomized controlled study of prostaglandin 15-methyl F2 alpha compared with syntometrine for prophylactic use in the third stage of labour. AB - A randomized controlled study of 112 women with singleton pregnancies at term, and no antenatal complications, admitted in spontaneous labour were randomized to receive either an intramuscular injection of 0.5 mg of Syntometrine or an intramuscular injection of 125 ug of prostaglandin 15-methyl F2 alpha at delivery of the anterior shoulder of the baby. Blood lost in the first 2 hours, and subsequent 22 hours postdelivery were collected separately and measured by colourimetric measurement of haemoglobin content. Other parameters in the third stage were measured, including need for transfusion of blood or blood products, length of the third stage, and change in haemoglobin concentration before and 24 hours after delivery. The incidence of side-effects with administration of either prostaglandin 15-methyl F2 alpha or Syntometrine were documented. The prophylactic use of intramuscular prostaglandin 15-methyl F2 alpha (Carboprost) in the active management of the third stage of labour gave similar results to prophylactic intramuscular Syntometrine in terms of length of the third stage of labour, incidence of postpartum haemorrhage and total blood loss in the first 2 hours and subsequent 22 hours after delivery. However it has the disadvantage of higher cost, as well as statistically significant increase in the incidence of profuse and frequent diarrhoea. Based on these results intramuscular injection of prostaglandin 15-methyl F2 alpha offers no advantage over intramuscular Syntometrine for routine prophylactic use to reduce blood loss in the third stage of labour. PMID- 8717568 TI - Efficacy of bupivacaine delivered by wound catheter for post-Caesarean section analgesia. AB - A prospective, randomized, double-blind trial was conducted to assess contribution to postoperative analgesia of intermittent instillation of 0.25% bupivacaine beneath the rectus sheath in 70 women delivered by lower uterine segment Caesarean section. The operations were performed via a Pfannenstiel incision under spinal anaesthesia. Background intravenous narcotic analgesia was provided with a patient controlled analgesia system (PCAS) using a standard morphine regimen. Overall (44 hr) mean morphine consumption was significantly greater in the placebo (saline) group compared to the treatment group (84.2 mg versus 63.3 mg. Two tailed t test p < 0.001). The most significant intergroup differences in narcotic use were found in the first 4 hours and between 24 and 36 hours after commencing PCAS (Two tailed t test p = 0.014 and 0.003 respectively). Subjective pain scores were assessed with a 10 cm visual analogue scale (VAS). The mean peak VAS score was greater in controls (5.37) than the treatment group (4.25) between 18 and 24 hours postoperatively (Mann-Whitney U = 424, p = 0.027). There were no intergroup differences in pain scores for any other time period. The overall incidence of nausea was lower in the treatment group compared to the control group (Chi squared with Yates' correction p = 0.046) and a lower degree of sedation was seen in those receiving bupivacaine between 4 and 8 hours after commencing PCAS (Mann-Whitney U = 427, p = 0.028). No differences in other narcotic related side-effects (vomiting and pruritus) were shown between groups. Regular instillation of 0.25% bupivacaine beneath the rectus sheath of women delivered by Caesarean section reduces their morphine requirements by 25% in the 44 hours after operation, with an associated reduction in both nausea and early sedation. PMID- 8717569 TI - Terminations of pregnancy for exposure to oral retinoids in South Australia, 1985 1993. AB - The oral retinoids, isotretinoin and etretinate, are highly teratogenic drugs which have been available in Australia since 1985 because of their unique effectiveness in severe cystic acne, psoriasis and other keratinization disorders. Only dermatologists can prescribe them, but in spite of strict guidelines by the manufacturers and the College of Dermatologists, exposed pregnancies have occurred. We attempted to determine the circumstances of exposure to these drugs for the 18 pregnancies terminated in South Australia in 1985-1993, using questionnaires to medical practitioners who notified the terminations under legislation. The main reason for their occurrence was the lack of compliance with the use of effective contraception by the women. In South Australia, some prescriptions of oral retinoids by unauthorized doctors have been dispensed during this period. Using statistics on prescriptions dispensed, an estimate was made of 1 termination of pregnancy for 319 courses of treatment for women with isotretinoin. Extension of guidelines for prescription and more detailed counselling in relation to the use of contraception are recommended. It is also suggested that doctors intending to recommend terminations obtain consent to discuss exposure with the dermatologists concerned, who may not be aware of the exposed pregnancy. PMID- 8717570 TI - Vertical transmission of HIV-1 in mid-trimester gestation. AB - Between July, 1994 and March, 1995, 23 heart blood samples from fresh abortuses of HIV-1 seropositive pregnant women after elective termination of pregnancy between 18 and 25 weeks of gestation by prostaglandin E1 analogue vaginal administration were examined for polymerase chain reaction (PCR) of HIV-1 genome and p24 antigen to investigate the transplacental transfer of HIV-1 infection. All samples of fetal heart blood were positive for HIV-1 antibody (ELISA), but negative for PCR and HIV-1 p24 antigen assay. These negative results could be due to the lack of the virus in the peripheral blood or to a viral load low enough to be undetectable by PCR method at mid-trimester gestation and suggest that HIV-1 vertical transmission occurs mostly during the last trimester of pregnancy and/or at delivery. PMID- 8717571 TI - HIV infection presenting as chronic postpartum perineal sepsis. PMID- 8717572 TI - Perforated duodenal ulcer-an unusual and often forgotten cause of an acute abdomen in pregnancy. PMID- 8717573 TI - A rare cause of an acute abdomen in late pregnancy. AB - A case is presented where spontaneous rupture of a uterine arteriovenous malformation into the peritoneal cavity presented as an acute abdomen with fetal distress in late pregnancy. This is thought to be the first reported case of its type. PMID- 8717574 TI - Combined chemotherapy in the medical management of tubal pregnancy. AB - The medical management of early unruptured tubal ectopic pregnancies is gaining acceptance internationally as an alternative to surgical procedures. This method has been shown to be effective and safe in properly selected cases and with adequate supervision. Most of the work however has been done using either methotrexate or prostaglandins. The present study aimed to evaluate the efficacy of a combined therapeutic regimen. The efficacy was noted to be at least as good but with fewer side-effects. The treatment was effective in 18 out of 19 cases of tubal pregnancies (94.7%). One patient complained of a gastritis which resolved with antacids. One patient experienced abdominal cramps and transient hypotension probably as a profound vagal response during tubal abortion. The median time to resolution varied directly with the initial serum beta HCG level at diagnosis. All patients who responded to the therapy described the experience as painless and viewed the treatment positively as it spared them the need for surgery and its attendant anaesthetic risks. However, we strongly recommend caution and close supervision and to keep the patient in the hospital at least for the first week of therapy. PMID- 8717575 TI - The human papillomavirus and the lower genital tract. PMID- 8717577 TI - A randomized comparison of the 3 Papanicolaou smear collection methods. AB - The purpose of this study was to compare the effectiveness of 3 cervical smear sampling devices for obtaining acceptable yields of endocervical cells in Papanicolaou smears. In a randomized, controlled clinical trial, we collected 346 smears over a 6-month period, 110 with the Cervex brush, 125 with the cotton swab plus Ayre spatula and 111 with the Cytobrush plus Ayre spatula. The Cytopathology Laboratory, blinded to the Papanicolaou smear method, screened the smears using the Bethesda system guidelines. Statistical analyses were performed using the Pearson X2 and analysis of variance tests. There was an increased detection of endocervical cells in the Cytobrush plus Ayre spatula and the Cervex brush methods, compared with the cotton swab plus Ayre spatula group (p = 2.39 x 10( 6), p = 4.49 x 10(-7) respectively). There was no statistically significant difference between the Cytobrush-Ayre spatula and the Cervex brush sampling methods were equally effective in obtaining endocervical cells in Papanicolaou smears. PMID- 8717576 TI - The significance of mild squamous atypia on cytology. AB - Between January, 1991 and February, 1993 inclusive, 396 Papanicolaou smears were reported to show Mild Squamous Atypia with or without Human Papilloma Virus (MSA +/- HPV). All women with MSA +/- HPV smears were routinely recalled for colposcopy. To determine the significance of MSA +/- HPV on routine smear screening, the records of all patients were reviewed. Three hundred and thirty seven women (85.1%) attended the colposcopy clinic and are the subjects analyzed for this report. The remaining 59 (14.9%) failed to attend. Intraepithelial neoplasia was found in 61 patients (18.1%), of whom 27 (8.0%) had a high grade lesion (CIN 2 or 3 or GIN 2). No patient had invasive cancer of the cervix. Only 1 of the 45 pregnant women had a significant lesion. It is concluded that all asymptomatic women with MSA +/- HPV on cervical smear may be managed in accordance with the current NH and MRC recommendations (1) and have a repeat smear in 6 months and colposcopy if the abnormality persists at 12 months. PMID- 8717578 TI - Absence of fetal corpus callosum associated with intrapartum cardiotocographic nonreactivity. PMID- 8717579 TI - Cervico-isthmic pregnancy with placenta percreta ending in a livebirth. PMID- 8717580 TI - Thyrotoxicosis in pregnancy presenting as pancytopenia. AB - Delay in diagnosis of thyroid disease can result in morbidity and mortality. Thyroid disorders are not uncommon in the reproductive age group and therefore in pregnancy; recognition of both the common and the more unusual manifestations will optimize outcome. PMID- 8717581 TI - Uterine prolapse and acute renal failure in a Chinese patient. PMID- 8717582 TI - Incarcerated procidentia: a rare complication of carcinoma cervix. AB - A rare case of a huge carcinoma of the cervix leading to sudden uterovaginal prolapse and incarcerated procidentia, in a postmenopausal woman is presented. The woman presented as an emergency with severe distress. The problem was successfully managed by reducing the uterovaginal prolapse under deep sedation and starting radiotherapy as an emergency procedure at the same sitting. The role of immediate radiotherapy in the management of such an emergency situation is discussed. PMID- 8717583 TI - Vaginal carcinoma associated with haematocolpos: a case report and review of vaginal adenosis as a precursor lesion. PMID- 8717584 TI - Falciparum malaria in pregnancy. PMID- 8717585 TI - Caesarean section and instrumental delivery rates. PMID- 8717586 TI - Laparoscopically-assisted vaginal hysterectomy. PMID- 8717587 TI - Laparovaginal procedures replacing abdominal hysterectomy. PMID- 8717588 TI - Diagnostic laparoscopy with minilaparotomy. PMID- 8717589 TI - Is expected utility theory normative for medical decision making? AB - Expected utility theory is felt by its proponents to be a normative theory of decision making under uncertainty. The theory starts with some simple axioms that are held to be rules that any rational person would follow. It can be shown that if one adheres to these axioms, a numerical quantity, generally referred to as utility, can be assigned to each possible outcome, with the preferred course of action being that which has the highest expected utility. One of these axioms, the independence principle, is controversial, and is frequently violated in experimental situations. Proponents of the theory hold that these violations are irrational. The independence principle is simply an axiom dictating consistency among preferences, in that it dictates that a rational agent should hold a specified preference given another stated preference. When applied to preferences between lotteries, the independence principle can be demonstrated to be a rule that is followed only when preferences are formed in a particular way. The logic of expected utility theory is that this demonstration proves that preferences should be formed in this way. An alternative interpretation is that this demonstrates that the independence principle is not a valid general rule of consistency, but in particular, is a rule that must be followed if one is to consistently apply the decision rule "choose the lottery that has the highest expected utility." This decision rule must be justified on its own terms as a valid rule of rationality by demonstration that violation would lead to decisions that conflict with the decision maker's goals. This rule does not appear to be suitable for medical decisions because often these are one-time decisions in which expectation, a long-run property of a random variable, would not seem to be applicable. This is particularly true for those decisions involving a non-trivial risk of death. PMID- 8717590 TI - Why expected utility theory is normative, but not prescriptive. PMID- 8717591 TI - The strengths and limitations of expected utility theory. PMID- 8717592 TI - Is risk neutrality rational? PMID- 8717593 TI - Expected utility theory--is it normative or simply "practical"? PMID- 8717594 TI - Eccentric diagnostic tests: redefining sensitivity and specificity. AB - Diagnostic tests exist for which the current definitions of sensitivity and specificity are inadequate. Such tests are called eccentric to distinguish them from normal diagnostic tests, for which the standard definitions are satisfactory. It is shown that when the standard definitions are used with eccentric tests there will be an appreciable range of cutoff values over which the test has no discriminating power. This is demonstrated with ROC analysis and information theory. New definitions of sensitivity and specificity that are valid for both normal and eccentric tests are proposed. These definitions require the use of two cutoffs to distinguish test-positive from test-negative individuals. When the new definitions are used with eccentric tests, performance improves significantly and the anomalous regions of zero discriminating power disappear. In the limit of zero eccentricity the new definitions become identical to the old. PMID- 8717595 TI - Empirical Bayes combination of estimated areas under ROC curves using estimating equations. AB - A synthesis of the empirical Bayes method and the method of estimating equations is used to combine individual receiver operating characteristic (ROC) area estimates from different studies of the same diagnostic test into a single estimate. This single estimate represents the population mean from which individual areas under the ROC curves were sampled. The only data needed to carry out the method are estimated areas under the ROC curves and the corresponding standard errors. PMID- 8717596 TI - Validating literature-based models with direct clinical trial results: the cost effectiveness of secondary prophylaxis for PCP in AIDS patients. AB - OBJECTIVE: To compare literature-based estimates of the cost-effectiveness ratios of strategies for secondary prophylaxis of Pneumocystis carinii pneumonia (PCP) in AIDS patients with estimates obtained using data from a recent comparative clinical trial. DESIGN: A decision-analytic Markov model with data on drug efficacy and toxicity from both the medical literature and a national randomized clinical trial. Drug costs were from average wholesale prices. Discounted life expectancy, total direct medical costs, and cost-effectiveness were projected in dollars per year of life saved (YLS). SETTING: Hypothetical for the literature based model, then the clinical trial results from the multicenter AIDS Clinical Trials Group (ACTG Protocol 021). PATIENT POPULATION: Patients with AIDS and a prior episode of PCP. INTERVENTIONS: Strategies included no prophylaxis, TMP-SMX (160/800 mg) daily, or aerosolized pentamidine (300 mg) monthly. Patients experiencing major toxic reactions to either medication would cross over to the other agent. MAIN RESULTS: In the literature-based model no prophylaxis was associated with a projected life expectancy of 1.430 years, and total direct cost of $42,080. TMP-SMX increased life expectancy to 2.051 years and cost to $42,300; for aerosolized pentamidine life expectancy was 2.066 years and cost $43,960. TMP SMX had an incremental cost-effectiveness ratio of $350 per YLS compared with no prophylaxis; the incremental ratio for aerosolized pentamidine was $2,950 per YLS when compared with no prophylaxis, but rose to $110,880 per YLS compared with TMP SMX. When data from ACTG clinical trial 021 were utilized in the model, the incremental cost-effectiveness ratio for TMP-SMX compared with no prophylaxis was $720 per YLS; aerosolized pentamidine was not cost-effective, and was "dominated" by TMP-SMX because it was associated with higher costs and shorter life expectancy. CONCLUSIONS: Literature-based cost-effectiveness models are useful in developing health policy before clinical trials are completed. Clinical trial results, when available, can be used to validate and revise these models. For secondary PCP prophylaxis in AIDS patients, TMP-SMX is substantially more cost effective than aerosolized pentamidine. PMID- 8717597 TI - Proportional heuristics in time tradeoff and conjoint measurement. AB - The time-tradeoff (TTO) test is widely used to measure quality of life for different health states. Subjects are asked to equate the value of living a given period in an inferior health state to the value of living a shorter period in good health. Applications of TTOs have been criticized based on the fact that the value of future life duration is taken as the future life duration itself. The authors show that for a health state in which a subject does not want to live longer than a specified amount of time, subjects' responses do not comply with the assumption that the value of the period in inferior health is equated to the value of the shorter period in good health. Actually, preference reversals with respect to such a health state point to the use of a proportional heuristic in the TTO test. Comparisons of the TTO test in these subjects with category scaling and difference measurements also favor a proportional interpretation of the TTO test. In tests based on conjoint measurement, these subjects also appear to use a proportional heuristic. Consequences of the use of the TTO test and conjoint measurement in quality-of-life models are discussed. PMID- 8717598 TI - Justifying prenatal screening and genetic amniocentesis programs by cost effectiveness analyses: a re-evaluation. AB - A strength of cost-effectiveness analysis is the presentation of all relevant clinical options, variables, and outcomes and the placement of values on the outcomes. Despite success in many fields of health policy, cost-effectiveness analysts have failed to provide a complete evaluation of prenatal screening and genetic amniocentesis programs. Specifically, all published cost-effectiveness analyses of these programs at best only partially acknowledge the potential life of the aborted fetus. These incomplete evaluations not only violate some basic principles of cost-effectiveness analysis, they also produce conclusions that may be misleading. Ultimately, society must answer two basic questions regarding the use of cost-effectiveness to justify prenatal screening programs: Is cost effectiveness analysis an appropriate tool for the evaluation of these programs? If so, then what are the outcomes of interest? This paper first shows that the current literature does not present a complete evaluation of these programs. The ramifications of this incomplete evaluation are presented from several perspectives. Finally, given the problems associated with the use of cost effectiveness analysis, the article suggests that cost-effectiveness analysis is not yet an appropriate tool to justify prenatal screening or genetic amniocentesis programs. PMID- 8717599 TI - Modeling boolean decision rules applied to multiple-observer decision strategies. AB - A model that derives multiple-observer decision strategy ROC curves for boolean decision rules applied to binary decisions of two or three observers is presented. It is assumed that covert decision variables consistent with ROC models of observer performance underlie decisions and that readers' decision criteria are in a fixed relationship. The specific parameters of individual ROC curves and the correlational structure that describes interobserver agreement have dramatic effects upon the relative benefits to be derived from different boolean strategies. A common strategy employed in clinical practice, in which the overall decision is positive if any observer makes a positive decision, is most effective when the readers are of similar ability, when they adopt similar decision criteria, when interreader agreement is greater for negative than for positive cases, and when the individual ROC slope is <<1.0. Different multiple observer decision strategies can be evaluated using the model equations. A bootstrap method for testing model-associated hypotheses is described. PMID- 8717600 TI - Patient satisfaction with health care decisions: the satisfaction with decision scale. AB - Patient satisfaction measures have previously addressed satisfaction with medical care, satisfaction with providers, and satisfaction with outcomes, but not satisfaction with the health care decision itself. As patients become more involved in health care decisions, it is important to understand specific dynamics of the decision itself. The Satisfaction with Decision (SWD) scale measures satisfaction with health care decisions. It was developed in the context of postmenopausal hormone-replacement therapy decisions. The six-item scale has excellent reliability (Cronbach's alpha = 0.86). Discriminant validity, tested by performing principal-components analysis of items pooled from the SWD scale and two conceptually related measures, was good. Correlation of the SWD scale with measures of satisfaction with other aspects of the decision-making process showed the SWD scale was correlated most highly (0.64) with "decisional confidence," and least with "desire to participate in health care decisions" and "satisfaction with provider." The SWD scale predicts decision certainty in this study. Use in an independent study showed that the SWD scale was correlated with the likelihood of patients' intentions to get a flu shot. Further investigation in relation to other health decisions will establish the utility of the SWD scale as an outcome measure. PMID- 8717601 TI - Measuring heart patients' willingness to pay for changes in angina symptoms. AB - Willingness-to-pay (WTP) measures of the effects of changes in health on a person's welfare are more comprehensive than traditional cost-of-illness (COI) measures, but they are sometimes difficult to obtain. The authors investigated two approaches for measuring heart patients' WTP for changes in their angina symptoms. First, actual expenditures and perceived angina episodes avoided were used to infer an averting-behavior measurement of WTP. Second, a contingent valuation approach was used to ask direct WTP questions regarding a hypothetical medical treatment that could be purchased to avoid additional angina episodes. The results indicated that although negligible COI changes were expected with small changes in angina frequency, the subjects had significant WTP to avoid increases in angina. The average WTP to avoid additional angina episodes revealed by the averting-behavior questions was comparable to the directly-elicited WTP, providing a test of the validity of the contingent-valuation approach. PMID- 8717602 TI - The relationship between cardiac functional capacity and patients' symptom specific utilities for angina: some findings and methodologic lessons. AB - Forty-one angina patients with coronary disease were interviewed to examine the correlation between prespecified and individualized weights for disease-specific measures of the effects of angina on the patients' well-being. Modifications of the Canadian Cardiovascular Society (CCS) scale for angina and the Duke Activity Specific Index (DASI) were used to rate functional capacity with prespecified items. Disease-specific utilities based on descriptions of functional status were obtained directly, and again indirectly with different anchoring conditions to control for noncardiac comorbidity. Correlations between the functional-capacity measures and the derived utilities were not strong, ranging from -0.25 (p > 0.1) to -0.35 (p = 0.02). Correlation between the two prespecified measures was higher (r = -0.51 or -0.69, both p < 0.01, for DASI versus CCS graded from walking and stair-climbing, respectively). The direct and indirect disease-specific utility scores were similar (r = 0.92, p < 0.01). The method described provides an approach to measuring disease-specific utilities by adapting existing scales for use in a standard gamble. It confirms that prespecified functional status scores inconsistently reflect patients' valuations of functional states. Further investigation should address whether the observed null effect of comorbidity on disease-specific utilities arose from inadequate coverage of the comorbidity issues in patient interviews or from lack of power given the small size of the feasibility study. PMID- 8717603 TI - The college student, the dentist, and the North Carolina senator: risk analysis and risk management of HIV transmission from health care worker to patient. AB - The risk of acquiring human immunodeficiency virus (HIV) infection from a health care worker is 2,000 times less than that of dying from a car accident. It is 700 times less probable than perishing from being struck by lightning or suffering a fatal fall. Despite the rarity of this occurrence, reduction of the risk of health-care-worker-to-patient HIV transmission in the workplace has been the focus of congressional, federal, state, and local agencies. If all HIV transmission from health care workers to patients were prevented using current guidelines and legislation, the epidemic of AIDS would be reduced by 0.0006%. Current efforts to prevent HIV transmission from health care workers to patients are the result of incomplete risk analysis and management. In a society of limiting resources and of cherished freedoms, sanctions imposed on health care workers to prevent HIV transmission to patients may benefit no one. PMID- 8717604 TI - Neuropeptide deficits in schizophrenia vs. Alzheimer's disease cerebral cortex. PMID- 8717605 TI - Neuropeptide deficits in schizophrenia vs. Alzheimer's disease cerebral cortex. AB - Neuropeptide concentrations were determined in the postmortem cerebral cortex from 19 cognitive-impaired schizophrenics, 4 normal elderly subjects, 4 multi infarct dementia (MID) cases, and 13 Alzheimer's disease (AD) patients. Only AD patients met criteria for AD. The normal elderly and MID cases were combined into one control group. Somatostatin concentrations were reduced in both schizophrenia and AD. Neuropeptide Y concentrations were reduced only in schizophrenia, and corticotropin-releasing hormone concentrations were primarily reduced in AD. Concentrations of vasoactive intestinal polypeptide and cholecystokinin also were reduced in schizophrenia, although not as profoundly as somatostatin or neuropeptide Y. In AD, cholecystokinin and vasoactive intestinal peptide were unchanged. Neuropeptide deficits in schizophrenics were more pronounced in the temporal and frontal lobes than in the occipital lobe. The mechanisms underlying these deficits in schizophrenia and AD are likely distinct. In schizophrenia, a common neural element, perhaps the cerebral cortical gaba-aminobutyric acid (GABA)-containing neuron, may underlie these deficits. PMID- 8717606 TI - Symptom stability in geriatric chronic schizophrenic inpatients: a one-year follow-up study. AB - The results of previous studies of symptom stability in schizophrenia suggest that negative symptoms manifest traitlike characteristics while positive symptoms fluctuate over time. Various prospective studies of chronic schizophrenic patients have found consistent results, regardless of the follow-up period, yet there is little research addressing symptomatology in geriatric schizophrenic patients. Since these patients have a very poor outcome and more severe negative symptoms, their symptoms might differ from younger patients. This study examined the course of symptomatology in 178 geriatric schizophrenic inpatients who were assessed twice at a 1-year interval with the Positive and Negative Syndrome Scale (PANSS). Intraclass correlations revealed that the distribution of negative symptoms was considerably more stable than that of positive symptoms over the interval, and subtypes based on negative symptoms were the only ones that manifested consistent stability over time. There was also a significant increase in negative symptom severity for the sample, with a slight decrease in positive symptom severity. Thus, even in chronic inpatients, with a very extended illness, positive symptom severity is not particularly stable within patients. These data indicate that the characteristics of negative and positive schizophrenic symptoms are similar in younger and geriatric schizophrenic patients, suggesting a continuity of the illness process. Tentative evidence for increasing severity of negative symptoms over a brief follow-up period suggests the possibility of a steady worsening of clinical state in very elderly patients who remained hospitalized. PMID- 8717607 TI - Brain serotonin (5-HT) neuroendocrine function in patients taking cholesterol lowering drugs. AB - Brain serotonin (5-HT) neuroendocrine function, plasma tryptophan, and platelet 5 HT content were examined in 20 patients treated in a lipid clinic for hypercholesterolaemia with combined drug and diet therapy and in 20 healthy matched controls. Treatment had produced a substantial decrease in total cholesterol concentrations in the patients, but they still had higher cholesterol and triglyceride levels than control subjects. The patients were somewhat more depressed than controls but did not differ from them in degree of hostility, free or total plasma tryptophan, or prolactin response to 30 mg of d-fenfluramine. This study does not reveal evidence of abnormal brain 5-HT neuroendocrine function in hypercholesterolaemic patients receiving cholesterol-lowering medications and diet. PMID- 8717608 TI - [123I]IBZM SPET analysis of dopamine D2 receptor occupancy in narcoleptic patients in the course of treatment. AB - Elevated levels of central D2 dopamine receptors were found on postmortem examination in cases of human narcolepsy. In vivo investigations using positron emission tomography (PET) and single photon emission tomography (SPET) found no changes of D2 binding in the striatal structures. To investigate whether the elevated D2 receptors in postmortem investigations are due to long-term treatment effects, we applied 123I-labeled (S)-2-hydroxy-3-iodo-6-methoxy-([1-ethyl-2 pyrrolidinyl]methyl) benzamide (IBZM) ([123I]IBZM, a highly selective CNS D2 dopamine receptor ligand) and SPET in narcoleptic patients in the course of treatment with stimulants and/or antidepressants. Before treatment we found no changes in D2 binding in 10 patients (in comparison to 10 normal controls). After treatment (performed in five patients for 3 months) we found changes in D2 binding in four of them, indicating that the results of the postmortem studies could have been influenced by long-term medications. Human narcolepsy seems not to be related to a striatal D2 dopaminergic disturbance. PMID- 8717609 TI - Fewer pigmented locus coeruleus neurons in suicide victims: preliminary results. AB - Studies of the noradrenergic system in suicide victims have found evidence of alterations in cortical beta- and alpha-adrenergic receptor binding. Since these receptor changes may be secondary to altered noradrenergic input, we sought to determine whether the pigmented neurons of the locus coeruleus (LC), which provide the noradrenergic innervation to the cerebral cortex, are altered in suicide victims. We studied 11 controls without known psychiatric or neurologic disorders and six suicide victims. LC neuron number, LC volume, and neuron density were determined by computer-assisted mapping. The suicide group had 23% fewer LC neurons and a 38% lower density of LC neurons than controls. The reduction in neuron number was localized to the rostral two thirds of the LC. Neither the LC length nor the LC volume in suicide victims differed from controls. Altered brain noradrenergic neurotransmission in suicide victims may be due to fewer noradrenergic neurons in the LC. Further studies are needed to determine whether this noradrenergic neuron loss is associated with an underlying major depression or specifically with suicidal behavior. PMID- 8717610 TI - Effects of dietary selenium on mood in healthy men living in a metabolic research unit. AB - Eleven healthy men were confined in a metabolic research unit for 120 days in a double-blind study of the effects of dietary selenium on mood as assessed by the Profile of Mood States-Bipolar Form. At an intake of 2800 kcal/day, the diet of conventional foods provided 80 micrograms/day of selenium for the first 21 days, then either 13 or 356 micrograms/day for the remaining 99 days. There were no significant changes in any of the mood scales due to dietary selenium. However, in the low-selenium group, the changes in the agreeable-hostile and the elated depressed subscales were correlated with initial erythrocyte selenium concentration; that is, the lower the initial selenium status, the more the mood scores decreased. These results suggest that persons with low selenium status might experience relatively depressed moods and support the idea that selenium plays a special role in the brain. However, these studies do not support the notion that selenium supplementation could promote improvements in mood in persons eating a typical U.S. diet. PMID- 8717611 TI - A behavioral animal model of posttraumatic stress disorder featuring repeated exposure to situational reminders. AB - The purpose of this study was to evaluate an animal model of posttraumatic stress disorder (PTSD) in mice. The model featured repeated exposures to situational reminders of a traumatic stress, which consisted of a brief electric shock, and included assessment of two behavioral parameters and the startle reflex. The findings indicated an initial, but unsustained, increase in locomotor activity in a neutral environment due to traumatic stress. Exposure to situational reminders was associated with a persistent bidirectional abnormal behavioral pattern in a fear-provoking environment and a progressive increase over time in the magnitude of the startle reflex. Exposure to situational reminders also produced an increase in aggressive behavior. This animal model appears to produce behavioral changes analogous to those seen in patients with PTSD. PMID- 8717613 TI - Revisiting the Ehlers and Kupfer hypothesis: the growth hormone cortisol secretion ratio during sleep is correlated with electroencephalographic slow wave activity in normal volunteers. PMID- 8717612 TI - Clozapine withdrawal effects and receptor profiles of typical and atypical neuroleptics. AB - Withdrawal effects of neuroleptics have not received much attention. Clozapine withdrawal phenomena have been attributed to psychosis arising from D2 supersensitivity, which is unlikely since it has minimal action on D2 receptors. The time course and clinical features of this phenomenon suggest that cholinergic overdrive and gamma-aminobutyric acid (GABA) supersensitivity occurs after withdrawal, since it is strongly anticholinergic and has a GABAergic action. Recently, a number of patients showed marked decompensation when they were switched from clozapine to risperidone, especially when they were rapidly tapered off clozapine. This was probably more due to withdrawal effects than the primary psychosis or a lack of efficacy of risperidone. A slow withdrawal schedule would facilitate homeostatic mechanisms; anticholinergics would be useful in clozapine withdrawal. This area has not received any attention from researchers, nor are there any guidelines for clinicians. This will be particularly important with the widespread use of atypical agents in the future. PMID- 8717614 TI - Attention-related brain potential and cognition in alcoholism-associated organic brain disorders. PMID- 8717616 TI - Pilot study of hirsutism in women with psychiatric disorders. PMID- 8717615 TI - Gender differences in response to intranasal cocaine administration to humans. PMID- 8717617 TI - Chronic rejection: is allograft destruction an inevitable phenomenon? PMID- 8717618 TI - Immune mechanisms of long-term graft acceptance. PMID- 8717619 TI - Potential involvement of growth factors in the proliferation of arteriolar smooth muscle cells during chronic graft rejection. PMID- 8717620 TI - Nonimmunologic factors involved in long-term renal allograft deterioration. PMID- 8717621 TI - Discovery, development, and clinical trial strategies for immunosuppressive agents. PMID- 8717622 TI - Relapse of focal segmental glomerulosclerosis after kidney transplantation. PMID- 8717623 TI - Donor-specific bone marrow cells suppress lymphocyte reactivity to donor antigens and differentially modulate TH1 and TH2 cytokine gene expression in the responder cell population. PMID- 8717624 TI - The long walk toward the PKD1 gene. The European PKD1 Consortium. PMID- 8717625 TI - Autosomal recessive polycystic kidney disease: clinical features and genetics. PMID- 8717626 TI - Adhesion molecules and acute renal failure. PMID- 8717627 TI - Prevention of experimental acute tubular necrosis: current clinical applications and perspectives. PMID- 8717628 TI - TH1 and TH2 lymphocytes in autoimmunity. PMID- 8717629 TI - A Ca(2+)-sensing receptor: from physiology to inherited disorders of calcium homeostasis. PMID- 8717630 TI - Aquaporins: from physiology to nephrogenic diabetes insipidus. PMID- 8717631 TI - The renal epithelial sodium channel: new insights in understanding hypertension. PMID- 8717632 TI - In vivo studies of parathyroid gland function in secondary hyperparathyroidism. AB - In contrast to studies in vitro, in vivo tests of parathyroid gland function in human subjects with secondary hyperparathyroidism present a number of technical and theoretical difficulties. The four-parameter model was originally developed to characterize the secretion of PTH in vitro by a defined number of parathyroid cells in tissue culture, whereas comparisons among in vivo studies in patients with varying degrees of parathyroid gland hyperplasia may not be feasible with this model. Maximum serum PTH levels during hypocalcemic stimuli in vivo reflect not only the amount of PTH released by each parathyroid cell but also the total number of cells available for hormone secretion. Parathyroid gland size varies markedly in patients with secondary hyperparathyroidism, and there is currently no reliable technique for accurately measuring the amount of parathyroid tissue present. Accordingly, in vivo comparisons of calcium-regulated PTH release among subjects with parathyroid glands of substantially different sizes may not be valid with the four-parameter model. Kwan et al. have suggested that parathyroid gland function be assessed over a wide range of different calcium concentrations. In this model, the set point for calcium-regulated PTH secretion is not calculated, but the relationship between serum ionized calcium and PTH levels is examined by linear regression after semilog transformation of the data. Linearizing the calcium-PTH curve eliminates the need for more complex curve fitting analyses. Differences in parathyroid gland function have been reported before and after calcitriol therapy when this approach is used, but the severity of secondary hyperparathyroidism varied widely in the patients evaluated. Alternatively, the Cica-clamp technique to quantify PTH secretion has been developed by Schwarz et al. By employing sequential standardized hypocalcemia and hypercalcemia, set point values obtained with this method closely agree with in vitro data obtained by Brown in normal parathyroid tissues. Advantages of the Cica-clamp technique include a shorter duration of study, reproducibility of the method, and absence of adverse effects; its utility in evaluating patients with either primary or secondary hyperparathyroidism remains to be determined. Based on data currently available, separate assessments of the pattern of change in serum PTH levels during hypocalcemia and hypercalcemia, expressed as the percent change from baseline values (see Fig 4), may provide more useful information about disturbances in the regulation of PTH release by calcium in patients with secondary hyperparathyroidism. Nevertheless, alterations in the set point for calcium-mediated PTH secretion do not adequately explain the excess PTH secretion of patients with secondary hyperparathyroidism, nor do they account for the reduction in serum PTH levels after treatment with calcitriol. PMID- 8717633 TI - Advanced glycosylation in nephropathy of diabetes and aging. PMID- 8717634 TI - Can cardiovascular complications be prevented in dialysis patients? PMID- 8717635 TI - An analysis of contemporary morphological concepts of synaptic remodelling in the CNS: perforated synapses revisited. AB - Perforated synapses refer to a synaptic type found in the central nervous system. They are characterized by their large size and by a discontinuity of the postsynaptic density when viewed in transverse sections, and by a doughnut or horseshoe shape when viewed in en face views. Of recent morphological studies, one approach has followed their characteristics throughout development and maturity, while others have concentrated on their probable roles in activities including kindling, long-term potentiation, spatial working memory, differential rearing, and the functioning of neuroleptics. An assessment is made of the hypotheses and models that have proved determinative in the emergence of perforated synapses as being significant in synaptic plasticity. Their distribution and frequency are summarized, with emphasis on the importance of unbiased stereological procedures in their analysis. Using three-dimensional approaches various subtypes are recognized. Of these, a complex or fragmented subtype appears of especial significance in synaptic plasticity. Ideas regarding the life-cycle of perforated synapses are examined. The view that they originate from conventional, non-perforated synapses, enlarge, and subsequently split to give rise to a new generation of non-perforated synapses, is critically assessed. According to an alternative model, perforated and non-perforated synapses constitute separate populations from early in their development, each representing complementary forms of synaptic plasticity. An attempt is also made to discover whether synaptic studies on the human brain in normal aging and in Alzheimer's disease throw light on the role of perforated synapses in synaptic plasticity. The loss of synapses in Alzheimer's disease may include a loss of perforated synapses - of particular relevance for an understanding of certain neuropathological conditions. PMID- 8717636 TI - GABAergic deafferentation hypothesis of brain aging and Alzheimer's disease; pharmacologic profile of the benzodiazepine antagonist, flumazenil. AB - Recent experiments have shown that: 1) A chronic 10 month daily administration to rats of the benzodiazepine (BDZ) receptor antagonist, flumazenil (FL; 4 mg/kg in drinking water), from the age of 13 through 22 months, significantly retarded the age-related loss of cognitive functions, as ascertained by the radial arm maze tests conducted two months after FL withdrawal. 2) An equal number of 8 rats died in the control and FL-treated group before the behavioral tests were completed and the animals were sacrificed; the life span of the FL-treated 8 rats equaled 24.0 (+/- 0.6 SEM) months, while that of the control 8 rats equaled 22.3 months (+/- 0.7 SEM), and the group difference was marginally significant (p = 0.04 Mann Whitney test). 3) In rats sacrificed 3 months after FL withdrawal and behavioral testing, the protective action of FL, relative to age-matched controls, was revealed by a significant reduction in the age-related loss of neurons in the hippocampal formation. 4) In the time period of 3 months between the drug withdrawal and sacrificing of the animals, stress experienced by the aging rats during behavioral testing, related to excessive daily handling of the animals and partial food deprivation to motivate them to perform in the radial arm maze, apparently had excitotoxic effects on the hippocampal neurons, as indexed by the presence of 30% neurons in a state of moderate pyknosis found both in the FL group and the age-matched controls. In the 6 months "young" control group, the number of pyknotic neurons equaled only 3.5%. It was concluded that the drug withdrawal and stress of behavioral testing unleashed the previously FL controlled age-related degeneration. On the basis of these results and the literature, showing that the tone of the GABAergic system increases with age, and particularly in Alzheimer's disease (AD), the hypothesis of brain aging was formulated. It postulates that in mammals, with growing age, and prematurely in humans with AD, the increasing tone of the BDZ/GABAergic system interferes with antero- and retrograde axonal transport through a chronic depolarizing block of preterminal axon varicosities of the ascending aminergic and cholinergic/peptidergic systems, which are indispensable for normal metabolic/trophic glial-neuronal relationships. Such a state leads to discrete anatomic deafferentation of forebrain systems, and particularly of the neocortex, where block of the anterograde axonal transport results in induction of the cortical mRNA responsible for synthesis of the beta-amyloid precursor protein (beta APP). The simultaneous block of retrograde transport from chronically depolarized preterminal axon varicosities may account for toxic accumulation in cortex of the nerve growth factor (NGF) and other trophins, without which the basal forebrain cholinergic neurons degenerate. The general pharmacologic profile of FL has been discussed on the basis of FL administration to animals and healthy and diseased humans. This profile shows that FL: 1) increases brain metabolic functions; 2) reduces emotional responses, thereby stabilizing the functions of the autonomic system in both humans and animals challenged by adverse environmental stimuli; 3) improves cognitive and coordinated motor functions in both humans and animals; 4) uniquely combines anxiolytic, vigilance and cognitive enhancing, i.e. nootropic, properties, which may, in part, stem from FL-induced emotional imperturbability (ataraxy); 5) facilitates habituation of healthy humans and animals to novel but inconsequential environmental stimuli, and promotes non-aggressive interactions among animals; 6) in single i.v. doses, and administered chronically to humans, FL has antiepileptic actions in the Lennox Gastaut syndrome and other forms of epilepsy characterized by "spike-and-dome" EEG patterns; these actions are likely to depend on FL's disinhibition of the serotonin system; 7) administered in single i.v... PMID- 8717637 TI - Long-term depression: a learning-related type of synaptic plasticity in the mammalian central nervous system. AB - Studies of various forms of synaptic plasticity in the central nervous system have provided insights into the cellular and molecular mechanisms for certain types of learning and memory. Activity-induced decreases and increases in synaptic efficacy can be elicited in mammalian neurons. Long-term depression (LTD) and long-term potentiation (LTP) are two major forms of activity-dependent synaptic plasticity in the brain. LTD of excitatory synaptic transmission in the cerebellum in the most well studied form of synaptic depression. The induction of cerebellar LTD requires conjunctive activation of alpha-amino-3-hydroxy-5-methyl 4-isoxalepropionate (AMPA) receptors, metabotropic glutamate receptors (mGluRs) and L-type voltage-dependent Ca2+ channels. Several intracellular second messengers and protein kinases are critical for cerebellar LTD, including cGMP, cGMP-dependent protein kinase and protein kinase C (PKC). A novel intercellular messenger, nitric oxide (NO), is found in the cerebellum, is released durinng synaptic stimulation, and may contribute to cerebellar LTD. The expression of cerebellar LTD is mediated by postsynaptic desensitization of AMPA receptors. Recently, a form of homosynaptic LTD has been described in the CA1 region of the hippocampus. The induction of hippocampal LTD is postsynaptic. N-Methyl-D aspartate receptors and mGluRs are important for induction of hippocampal LTD. Other intracellular and intercellular messengers, such as NO, cGMP and cAMP, might act downstream from glutamate receptors during hippocampal LTD. The expression of hippocampal LTD is likely to be in part presynaptic. While cerebellar LTD may be important for motor learning, the behavioral role of hippocampal LTD remains to be explored. PMID- 8717639 TI - Heart failure from diastolic dysfunction. AB - Heart failure from diastolic dysfunction is a clinical syndrome similar to, but distinct from, failure from systolic dysfunction. Because the standard cardiac diagnostic tools may not be helpful, the advance practice nurse and physician collaborate on the diagnosis. Interventions are aimed toward improving diastolic filling and reducing hemodynamic compromise for these patients. PMID- 8717638 TI - Opioid receptors and the regulation of ion conductances. AB - In the past two years, knowledge about the regulation of ion conductances by opioid receptors has expanded in several directions. First, it has now been shown that all three of the major receptor subtypes act on all three of the well recognized effectors, i.e., adenylyl cyclase, calcium channels and potassium channels. Second, the opioid-mediated receptor inhibition of adenylyl cyclase has been linked to the modulation of ion channel activity /20/. Third, opioids have also been found to increase the activity of adenylyl cyclase in some preparations. Fourth, opioid receptors can mediate a rise in internal free calcium concentration /21/. These observations suggest that the regulation of cell excitability by opioids is dependent on the cell under study and that the generalization between receptor subtype and specificity of action is no longer valid. The additional second messenger pathways affected by opioids suggest that the adaptive changes resulting from chronic opioid treatment are more complex than previously thought. This review evaluates these relatively new observations and suggests how these results may change the interpretations from previous work on opioid actions. PMID- 8717640 TI - Aggressive weaning in cardiac surgical patients. AB - Nursing management of cardiac surgical patients has changed dramatically over the last several years. The need for cost-effective care with quality patient outcomes has forced critical care nurses to re-evaluate the standard of practice for postoperative cardiac patients. The authors describe the implementation of a fast-track weaning program giving critical care nurses more responsibility for weaning decisions and choice of sedative agent. Our experience in a six-bed Cardiothoracic Recovery Unit (CRU) in a midwestern community hospital evolved from receiving heavily anesthetized patients, continuing sedation postoperatively and prolonged intubation to minimal pharmacologic intervention with propofol and early extubation. In the fast-track weaning protocol, the nurses were given more authority to make critical decisions during the weaning process. This article describes the transition from the traditional time-based weaning system to the fast-track system where individual weaning progress is monitored by the critical care nurses. PMID- 8717641 TI - Cyclosporine and tacrolimus: a comparison of immunosuppressants used for liver transplantation. AB - The life expectancy of transplant patients has dramatically increased because of the development of cyclosporine to combat graft rejection. More recently, tacrolimus has been approved and is expected to further enhance survival. This paper compares these two medications and explores the role of the critical care nurse in monitoring outcomes and preventing side effects in patients with these immunosuppressants. PMID- 8717642 TI - Very early extubation: extubating in the OR following coronary artery bypass. AB - Cardiac surgery patients are being extubated earlier than ever before. This article describes how the nurses responded when the first patient to be extubated in the OR was admitted to the critical care unit. This case shows how this change is not as simple as just extubating the patient. Early extubation requires protocol changes, ethical considerations, system changes, staff education, and evaluation of patient outcomes. PMID- 8717643 TI - Rewarding clinical competence in the ICU: using outcomes to reward performance. AB - The rising acuity of hospitalized patients requires that critical care nurses maintain a high level of clinical expertise. Expert skill levels (critical thinking, technical skills, and interpersonal skills) of clinicians improve patient outcomes that translates into savings for the institution through shorter length of stay and decreased frequency of re-admissions. The authors outline the steps to follow in developing a theory-based career advancement plan that encourages and rewards clinical competence. PMID- 8717644 TI - Sexual counseling of MI patients: nurses' comfort, responsibility, and practice. AB - Myocardial infarction patients often have unanswered questions about resuming sexual activity after this life-threatening event. Critical care nurses can play a pivotal role in educating patients in this sensitive area. However, sexual counseling is an area of nursing that is frequently neglected. The authors assessed comfort and responsibility levels and their relationship to the practice of sexual counseling by critical care nurses. PMID- 8717645 TI - Health care rationing debate out in the open. PMID- 8717646 TI - The distress of circumcision. PMID- 8717647 TI - From ward sister to ward manager. PMID- 8717648 TI - Renal biopsy as a day case procedure. PMID- 8717649 TI - Clinical ladders: advancement for nurses. PMID- 8717650 TI - Paediatric hospital at home: the first year. PMID- 8717651 TI - A pictorial assisted teaching tool for families. PMID- 8717653 TI - The death of changing childbirth. PMID- 8717652 TI - Enteral nutrition. PMID- 8717654 TI - Smoking, mother's rights and the maternity department. PMID- 8717655 TI - Directed pushing in the second stage of labour. PMID- 8717656 TI - Midwifery degrees--will they deliver? AB - Midwifery education is in a process of change; links to higher education are being encouraged. Degrees should provide a greater theoretical base from which to practise. Degrees may improve practitioners' confidence, self-worth and negotiating skills. There are fears that graduates may become distanced from basic care provision. Perceptions exist that degree education is cut off from practice reality. Degree courses should be questioning, reflective, research based and woman-centred. PMID- 8717657 TI - Midwifery care for genitally mutilated women. PMID- 8717658 TI - Midwife unit of the year. PMID- 8717659 TI - Mother's own expressed breast milk: guide-lines for storage. AB - Guide-lines for milk collection and storage must aim to provide human milk with a composition as similar as possible to that stored within the human breast. The way in which milk is collected and stored has a significant effect on milk composition. Polyethylene storage bags should not be used to store milk for hospitalized infants. Glass containers fitted with an air-tight seal maintain immunoglobulin stability, and minimise fat loss. PMID- 8717660 TI - Cleft lip and palate. PMID- 8717661 TI - A student midwife's reflective diary. PMID- 8717662 TI - [Art therapy]. PMID- 8717663 TI - [Mask play. Created and played by adolescents hospitalized in a psychiatric department]. PMID- 8717664 TI - [Art therapy and studios for expression. Interview with Kim James]. PMID- 8717665 TI - [From one place to the next ... why not?]. PMID- 8717666 TI - [Art therapy and expression studios. An essai on art supporting care]. PMID- 8717667 TI - [The therapeutic "Marionette" studio. Dealing with aggression]. PMID- 8717668 TI - [Psychiatry in the prison milieu. Regional medical-psychological services]. PMID- 8717669 TI - [From you to me]. PMID- 8717670 TI - [Forgotten memories]. PMID- 8717671 TI - Call me what I am. PMID- 8717672 TI - Expandable esophageal wire stents. AB - Esophageal endoprostheses or stents have been used successfully for more than 100 years in patients with esophageal or gastroesophageal (GE) junction cancer. In recent years, many manufacturers have produced these stents in various styles and materials, but the most innovative design is the expandable wire mesh stent. At Virginia Mason Medical Center, The Schneider Wallstent (Minneapolis, MN) and the Wilson Cook Z stent (Winston-Salem, NC) have been placed in a series of patients with obstructing or fistulizing neoplasms of the esophagus, GE junction, stomach, lung, or mediastinum. The nursing care of these patients requires meticulous attention to airway maintenance and oral suction. Stents cannot be repositioned after total deployment; thus, the nurse plays an important role in assuring correct placement. During and after the endoscopy, the nurse must be alert to any signs of possible complications, including bleeding, perforation, dyspnea from tracheal compression, and stent malposition. Several case studies are presented in this article, noting the patients' histories, indications for stent placement, and nursing care pre-, intra-, and postprocedure. PMID- 8717673 TI - A case-control study to examine the influence of maternal perinatal behaviors on the incidence of Crohn's disease. AB - This case-control study, which examines maternal perinatal characteristics and behaviors during pregnancy, was an initial step to determine whether an association exists between breast feeding, oral contraceptive use, smoking habits during pregnancy, and the incidence of Crohn's disease. The population studied was mothers whose children were diagnosed with Crohn's disease before the age of 22. Population controls were selected by each case mother, who solicited two friends or neighbors with a child of similar age. Case mothers differed significantly on the variables of allergies (p = .05) and family history of inflammatory bowel disease (p = .001). No significant differences were found regarding feeding, oral contraceptives, or smoking. Using a forward step-wise logistic regression method, only family history remained in the equation. PMID- 8717674 TI - Living with ulcerative colitis. AB - This pilot study was undertaken to describe the experience of living with ulcerative colitis. Using a phenomenological approach, three subjects with diagnosed ulcerative colitis were interviewed and asked to describe what it was like to live with ulcerative colitis. Data from transcriptions of the audiotaped interviews were gathered and analyzed. Five major themes arose: the uncertainty surrounding exacerbations; the fear and humiliation surrounding stool incontinence; the desperate need to find successful treatment; the profound effect on family life, social life, and work; and, finally, the feeling of being controlled by the disease. Findings detail the impact of the disease on the subjects' lives and provide insight into this devastating disease. Implications for nursing practice and research are discussed. PMID- 8717675 TI - Do primary care providers screen for eating disorders? AB - This study was designed to evaluate the differences in nutritional assessments when performed by nurse practitioners, physicians, and physician assistants. Complete patient histories and physical records were assessed with the focus on the detection and prevention of eating disorders in female adolescents. Data included medical records of 111 female adolescents from a family practice with three separate sites. Primary care providers included two nurse practitioners, two physician assistants, and nine physicians. The null hypothesis that there is no difference in assessments of nurse practitioners, physician assistants, and physicians was rejected. A significant difference was found in the assessments of nurse practitioners and physicians (t = 2.62; p = .01). Nurse practitioners were found to be significantly more likely than physicians to perform assessment for eating disorders. PMID- 8717676 TI - GI nurses' retrospective look at foreign body ingestions in children. PMID- 8717677 TI - Flumadine (rimantadine hydrochloride). PMID- 8717678 TI - Radiation safety in the endoscopy setting. Society of Gastroenterology Nurses and Associates, Inc. PMID- 8717679 TI - Is versatility a by word for mediocrity. PMID- 8717680 TI - There are no difficult patients. PMID- 8717681 TI - Creating nursing links in our region. PMID- 8717682 TI - 'I'm on edge all the time': residents' experiences of living in an integrated nursing home. AB - This paper reports on interviews with six residents of an integrated nursing home who were asked: What is it like to live with others who exhibit socially unacceptable behaviours? Using a phenomenological approach, analysis elicited three major themes from participants' descriptions of their experiences: enervation, disenfranchisement and coping. Residents' comments highlighted the overwhelming efforts that their circumstances demanded of them; what these efforts meant to them is interpreted and the implications of the findings for nurses are discussed. PMID- 8717683 TI - Men's experiences during their partner's first pregnancy: a grounded theory analysis. AB - This paper reports on research conducted with 53 men who were attending antenatal classes with their partners who were pregnant for the first time. The men discussed their experience of pregnancy in focus groups that were run by male midwives who were also fathers. A grounded theory analysis of the men's discussions generated the categories of anxiety, ambivalence, adjustment, separation and need to know. There were two core categories, confusion, which was strongly expressed and development which was manifest in a minority of men. A descriptive narrative demonstrated that most first time fathers were confused as their relationship with their partner changed and that their roles in relation to the baby and other people were unclear. The men's sense of self was threatened and they responded negatively to the challenges they experienced and felt distanced from their partners. The findings have implications for clinical and educational services as they show that most men were alienated by the manner in which information was presented. The men also felt that services focused on their partner's labour and the birth of the child and neglected their greatest concerns, that is their changing identity, their relationships and their future role as fathers. PMID- 8717684 TI - The critical thinking abilities of RNs entering university courses. AB - The critical thinking abilities of groups of registered nurses (n 143) who were entering the post-registration Bachelor of Nursing or one of the four graduate diploma in nursing courses offered by Griffith University were assessed using the Watson-Glaser Critical Thinking Appraisal Test. The mean scores of the Bachelor of Nursing student group, none of whom had attended university, were significantly lower than the mean scores of the graduate diploma students who had no experience of tertiary education. There was not a significant difference in the mean scores of the group who were accepted into the graduate diploma courses on the basis of previous university studies and the group whose acceptance was based on prior learning in non-tertiary courses and/or clinical nursing experience. There were also no significant differences in the mean scores of groups of students entering the four graduate diploma courses. Some implications of these findings for students and for the development and implementation of curricula are discussed. PMID- 8717685 TI - Nursing children with disabilities: a conceptual framework for organising services. AB - This paper describes a framework for the organisation of acute care and community nursing services for children with disabilities and their families. The framework is based on primary health care principles and is designed as a guide for generalist nurses and specialist disability care givers. PMID- 8717686 TI - Professional development. Dementia: professional issues (continuing education credit). PMID- 8717688 TI - Online for information. PMID- 8717689 TI - Growing pains. PMID- 8717687 TI - Breaking imperial taboos. PMID- 8717690 TI - Equal opportunities. Room at the top? PMID- 8717691 TI - Equal opportunities. The view from the top. PMID- 8717692 TI - Equal opportunities. Senior office politics. PMID- 8717693 TI - Equal opportunities. Men's talk. PMID- 8717694 TI - Student nurses: novices or practitioners of brilliant care? AB - Student nurses are assumed to be novices in terms of traditional models of knowledge and skills development. The research described in this paper explored how relatively inexperienced students provide sophisticated care. The idea of 'brilliant care' is used to describe strategies used to improve the care of patients who are deemed difficult to care for. These strategies can contradict norms in the clinical area. The students therefore also had to learn to reconcile these conflicts. The findings of the research are used to challenge the accepted view of expertise in nursing practice. Students learn complex interpersonal skills quickly. The received view of expertise may therefore be excessively professionalised and technological. This paper is an adapted version of a paper presented at the RCN Research Advisory Group Conference in April 1995. PMID- 8717695 TI - Prostitutes can help prevent the transmission of HIV. AB - Social stigma and misconceptions about prostitution are barriers to promoting sexual health. This paper discusses the use of public health interventions to break down these barriers. It also attempts to offer some understanding of female prostitutes in particular and explain why they should not be driven underground. PMID- 8717696 TI - The case for shared training for nurses and doctors. AB - This paper reports on a study of the case for shared training by doctors and nurses for six practical procedures. It examines doctors' and nurses' views and makes recommendations for the development of training. PMID- 8717697 TI - Manic depression: do people receive adequate support? AB - The paper describes a national survey of the membership of the Manic Depression Fellowship. The demographic characteristics of those with manic depression indicated that they were similar to that of community surveys of people with the disorder. Of the respondents, 89% reported at least one hospital admission. The results highlight the need for awareness of manic depression among ward nurses and the importance of community mental health nurses in the assessment and monitoring of suicide risk, the provision of information and help with vocational rehabilitation. PMID- 8717698 TI - Mental health. The nurse's support role. AB - The role of the nurse in the management and treatment of manic depression comprises social, psychological and pharmacological interventions. Achieving a balance is crucial to the management of this serious disorder and nurses are ideally placed to help in this. PMID- 8717699 TI - Bandaging: an assessment of practices. PMID- 8717700 TI - Supervised discharge. PMID- 8717702 TI - Field day for cynics. PMID- 8717701 TI - Health education: power to the people. PMID- 8717703 TI - Boom time for flying pigs. PMID- 8717704 TI - Nutrition. Malnutrition in hospitals. PMID- 8717705 TI - Nutrition. Nutritional assessment. PMID- 8717706 TI - Know how: vitamins and minerals. Part One. Vitamins, minerals and trace elements fundamental to recovery. PMID- 8717707 TI - Nutrition. Nourishing knowledge. PMID- 8717708 TI - Writing for publication. PMID- 8717709 TI - The assistant's guide to radial keratotomy. AB - 1. The role of the ophthalmic assistant in radial keratotomy (RK) patient care involves three basic areas: patient preparation, suite preparation, and RK day. Complications as well as pros and cons of RK are covered. 2. A truly informed consent to RK must be achieved. 3. The key to managing refractive patients is preparation and organization. Every technician working with RK patients should know every detail of the RK practice. The patients must be prepared as well; there should be no real surprises during the postoperative course. PMID- 8717710 TI - Extended wear soft contact lenses: to market or not to market. AB - 1. Today, cosmetic extended wear lenses have numerous aliases; continuous wear, provisional extended wear, and flexible wear, all referring to the same products, but on a relative scale of wearing time. Cosmetic extended wear lenses are elective and differ from the medical necessity of extended wear that few patients require. Refractive surgery has made significant advances in the cosmetic extended wear lens market. 2. Despite growing conservatism, positive marketing kept growing the EW philosophy for almost a decade until the FDA laid down some new rules, including a 1-week maximum for overnight wear. Coincidentally, this mandate came with the advent of the disposable lens in 1989. Since then, 1-week EW with disposable contacts has proved to be a safer modality for cosmetic EW; however, the concern of corneal complications continues to decline the market. 3. Patients risk a microbial ulcerative keratitis secondary to cosmetic EW. If daily wear contact lens options or occasional part-time EW is not amenable to the patient, refer the patient to an experienced refractive surgeon. Today, patients have more healthful vision choices than ever before. PMID- 8717711 TI - Ten years of ophthalmic assistant training: a multicultural approach and overview. AB - 1. Metropolitan areas with a diverse population require reevaluation of training techniques. 2. Communication skills have to be reinforced by a multicultural approach followed by a hospital-based apprenticeship program. 3. Competence level is constantly being reassessed in workshops, student-mentor supervision, and frequent evaluations. PMID- 8717712 TI - Case study of an orbital screwdriver injury. AB - 1. A man struck himself in the right eye with a screwdriver. The initial diagnosis was probable medial wall fracture with medial rectus entrapment, but a forced auction test result was negative and the initial x-ray study of the orbit and sinuses was within normal limits. The patient was treated with topical antibiotics and observed closely. Subsequent exams revealed a complete, isolated right gaze palsy. 2. An axial computed tomography scan of the orbit revealed an area of a probable hematoma overlying the lateral rectus muscle. There was also a fracture of the greater wing of the spheroid. 3. One month after the injury, the patient had complete, spontaneous clearing of the diplopia and lateral gaze palsy. The patient has had no residual effects 3 years later. This care reinforces the idea that injuries such as this should be treated symptomatically and observed. PMID- 8717713 TI - Resources for the low-vision community. AB - 1. The NAVH serves, exclusively, an otherwise overlooked group of disabled people -the visually impaired who are not totally blind. Although its numbers increase dramatically each year, the low vision community is largely unrecognized by the public. 2. Parent self-help discussion groups were initiated in the early 60s to help parents who expressed feelings of frustration that they could accept their offspring more readily if the child were totally blind. The first session supplied the vital information needed to prepare the first publications ever written about partially seeing children. 3. The agency has expanded to include all ages and to supply information about vision and vision throughout the world. PMID- 8717714 TI - Materials and solutions: staying in charge. PMID- 8717715 TI - [Cryopreserved homografts. History and organizational scheme]. PMID- 8717716 TI - [Cryopreservation: principles and technique]. PMID- 8717717 TI - [Homograft. Taking care of the patient]. PMID- 8717718 TI - [Homograft. Operating suite. Taking care of the patient]. PMID- 8717719 TI - [Critical chronic ischemia of the lower extremities. Use of a homograft to save the extremity]. PMID- 8717720 TI - [Angiography]. PMID- 8717721 TI - [Non-invasive vascular explorations]. PMID- 8717722 TI - [Placing the patient in the operating room and complications]. PMID- 8717723 TI - [Anesthesia in pregnant women. Physiological changes and how to deal with them]. PMID- 8717725 TI - [The other maternity]. PMID- 8717724 TI - [The uterine cervix. Basis for pelvic lymphadenectomy]. PMID- 8717726 TI - [The other maternity. Maternal care: why?]. PMID- 8717727 TI - [The other maternity. Maternal units]. PMID- 8717728 TI - [The other maternity. A pluridisciplinary staff]. PMID- 8717729 TI - [The other maternity. The mechanism of psychiatric maternity]. PMID- 8717730 TI - [The other maternity. Signs of mothers problems]. PMID- 8717731 TI - [The other maternity. Diagnosis in maternology]. PMID- 8717732 TI - [The other maternity. Treatment in maternal care. Theory and practice of treatment of early maternal dysfunction]. PMID- 8717733 TI - [The other maternity. Therapeutic nursing relationship]. PMID- 8717734 TI - [The other maternity. The video-clinical approach]. PMID- 8717735 TI - [The other maternity. Work in collaboration]. PMID- 8717736 TI - [The other maternity. Words of a mother: "in maternal care they do not do psychiatry, they look at you"]. PMID- 8717737 TI - [Life emergencies in pediatrics. Protocols, a code word, teaching of teachers]. PMID- 8717738 TI - [Giving children confidence]. PMID- 8717739 TI - The question of eligibility. PMID- 8717740 TI - Genetics, chromosomes and the elderly patient. PMID- 8717741 TI - Escape to the past. PMID- 8717743 TI - Difficulties on site. PMID- 8717742 TI - In favour of mobility. PMID- 8717744 TI - As time goes by. PMID- 8717745 TI - Care of urinary catheters. PMID- 8717746 TI - Community care: the German experience. PMID- 8717747 TI - Perception is a funny thing. PMID- 8717748 TI - Stretched to breaking point. PMID- 8717749 TI - Shortfall in mental health nurses. PMID- 8717750 TI - Nursing care is health care. PMID- 8717751 TI - Unrecognized drug reaction. PMID- 8717752 TI - Documenting suicide risk. PMID- 8717753 TI - Myths & facts ... about myasthenia gravis. PMID- 8717754 TI - Handling your patient's hearing aid. PMID- 8717755 TI - Photosensitivity reactions. PMID- 8717756 TI - Helping families cope with death and dying. PMID- 8717757 TI - Acute confusion in the elderly. PMID- 8717758 TI - Action stat. Adverse reaction to benadryl. PMID- 8717759 TI - Sinus bradycardia. PMID- 8717760 TI - Dressing up for a change: chest tube insertion site. PMID- 8717761 TI - Beating the odds: a teenager's fight for survival. PMID- 8717763 TI - 50 top nutrition tips. PMID- 8717762 TI - Assessing edema. PMID- 8717764 TI - Self-test. Managing emergencies. PMID- 8717765 TI - Dealing with sexual harassment. PMID- 8717766 TI - Saying good-bye to Pepaw. PMID- 8717767 TI - The process of obstetric triage: management by certified nurse-midwives. AB - The roles for certified nurse-midwives are growing, changing, and becoming more varied with the reform of health care systems. Versatility of clinical knowledge in antepartum, intrapartum, postpartum, and gynecologic care along with advanced practice skills of assessment, diagnosis, and treatment make nurse-midwives ideal providers to perform triage for a busy obstetric service. With health care dollars more limited and more scrutinized than ever before, it becomes essential to prioritize patient needs and match them with the most appropriate resources. The article demonstrates the process of triage provided by certified nurse midwives in a high-volume university hospital setting. It describes management guidelines and admission criteria for the birth center, high-risk labor and delivery, and the observation unit. Examples of clinical evaluations and judgments as well as scenarios are included to illustrate how triage is unique and differs from care provided in other settings. PMID- 8717768 TI - Obstetric triage: selected clinical management strategies by nurse-midwives in an academic tertiary facility. AB - Within the perinatal literature, there are few reports regarding obstetric triage. The documented use of advanced practice nurses performing obstetric triage is noted in some centers. The article documents the use of nurse-midwives in performing obstetric triage in an academic tertiary facility. Selected clinical scenarios are presented, and clinical management skills are delineated. PMID- 8717769 TI - A review of prescriptive authority for nurse practitioners. AB - Nurse practitioners in many states continue to struggle to gain prescriptive authority. Independent prescriptive authority holds in a small number of states. In states that grant prescriptive privileges, nurse practitioners battle strict regulations and mandatory supervision by a physician. The article reviews prescriptive authority for practitioners, interpretation of legislation, barriers to prescribing practice, and current trends. PMID- 8717770 TI - Compensation models of certified nurse-midwives in clinical practice. AB - Nurse-midwives have been recognized for decades as clinically effective health care providers for infants and women. Little information exists, however, as to how these clinically effective professionals are compensated for their services. The article presents five compensation models for certified nurse-midwives (CNMs), demonstrating the financial benefits and cost of each using a hypothetical case study format and composite data from current CNM practices in the United States. From these templates, CNMs can apply their own financial and productivity data to construct a financial analysis of their productivity against their current financial compensation. PMID- 8717771 TI - Advanced practice nursing in the care of the high-risk infant. AB - Advanced practice nursing encompasses a range of roles and services. Tertiary care settings such as neonatal intensive care units provide a variety of opportunities for the advanced practice nurse, including direct care, case management, and developmental care. Primary care and community settings may also provide opportunities for advanced practice nursing in the care of high-risk infants. Challenges related to role implementation, control of practice, and reimbursement also exist, however. The article examines the evolution of advanced practice nursing in the care of the high-risk infant as well as current opportunities and challenges. PMID- 8717772 TI - Clinical nurse specialists: creating new programs for neonatal home care. AB - Specialty home care programs are making headlines in nursing. Clinical nurse specialists are taking the lead in designing and directing follow-up home care for infants in neonatal intensive care units. The article describes a home care program in a Texas community that utilizes neonatal intensive care unit nursing staff as the home care providers for these infants. PMID- 8717773 TI - Neonatal pain: from concept to research questions and a role for the advanced practice nurse. AB - The assessment and treatment of neonatal pain remain inconsistent in spite of scientific evidence and agreement among practitioners that neonates do experience pain. In fact, pain forms the basis for many subsequent maturational events and may mediate future responses to pain. The article outlines the concept of pain in the neonate, provides suggestions for future research, and discusses the instrumental role of the advanced practice nurse in the recognition and treatment of neonatal pain. PMID- 8717774 TI - Advanced practice nurses: change agents for clinical practice. AB - The article presents the role of advanced practice nurses as change agents in the clinical setting. Research utilization is needed to shape and form the basis of nursing practice. As change agents, the advanced practice nurses in one institution established a research-based minimal stimulation protocol for the very low-birth weight infant and implemented it using a video presentation. Knowledge of change theory, research utilization, and video production was necessary in making this project a success. PMID- 8717775 TI - [Transplantation of hematopoietic stem cells. The transplantation unit. History and function]. PMID- 8717776 TI - [Hematopoietic stem cell transplantation]. PMID- 8717777 TI - [Hematopoietic stem cell transplantation. Specific nursing care. From technique to personal relations]. PMID- 8717778 TI - [Transplantation of cultured hematopoietic cells. A sample of nursing research]. PMID- 8717779 TI - [A history of breaches. The nursing profession in psychiatry]. PMID- 8717780 TI - [Recognizing, evaluating and treating pain. A challenge for the year 2000]. PMID- 8717781 TI - [Nursing in nephrology]. PMID- 8717782 TI - [From poster to slate. An information novelty in the hospital]. PMID- 8717784 TI - [Development of health care in the country]. PMID- 8717783 TI - [Mobile team of palliative care. Role of the nurse in relation to hospital nurses]. PMID- 8717785 TI - [Oral antidiabetics: acarbose]. PMID- 8717786 TI - [Nursing viewpoints in children with adult respiratory distress syndrome]. PMID- 8717787 TI - [Neurodermatitis, the visible disorder]. PMID- 8717788 TI - [Course of disease after surgery for hypoplastic left heart with mitral valve atresia and aortic isthmus stenosis--report by the mother]. PMID- 8717789 TI - [Report of parents' experiences with an allergic child]. PMID- 8717790 TI - [Nursing diagnosis--its importance for autonomy in nursing]. PMID- 8717791 TI - [Are special gifts permitted as gratuities for nursing care?]. PMID- 8717792 TI - [Toxic and less toxic plants. 26]. PMID- 8717793 TI - The power of words. PMID- 8717794 TI - Learning to care: gender issues for male nursing students. AB - The following article is a description of one aspect of a phenomenological research study designed to investigate the lived experience of male nursing students as they learned to care as nurses. Data-collection strategies included paradigm case narratives and interviews. Data analysis was characterized by four major strategies: analysis, synthesis, criticism, and understanding. These strategies were used to identify meanings of the text of transcribed interviews and to generate interpretive commentary. Learning to care was described by the participants as a complex entity that incorporates the gender of the student, the patient, the teacher, and the nurse. As students progressed through the program, their experience of gender issues in learning to care was shaped by personal experiences, the expectations of a predominantly female faculty and nursing staff, and their evolving understanding of the ways of caring that are gender based. PMID- 8717796 TI - Theorizing oppression: implications for nursing research on violence against women. AB - The way in which oppression is theorized is critical to nursing research in general and nursing research on violence against women in particular. Violence is not just the aberrant behaviour of some men, or a simple issue of gender oppression; rather, it is a complex, pervasive social problem that arises from and is sustained by multiple sites of oppression, including race, class, and gender. Conceptualizations of violence that neglect analyses of power, oppression, agency, and resistance may serve to support and sustain violence; thus their use in nursing research can align nursing with structures that perpetuate violence and oppression. Theories of violence that are informed by complex analyses of power and oppression focus inquiry on the social causes of violence; require analysis of racism, sexism, and classism; and establish, as a goal of research, the countering of oppression by individuals, institutions, and the state. Analyzing oppression as simultaneity guides nursing research into violence against women toward social change, is applicable beyond the issue of violence, and can help nursing contribute to the eradication of the social causes of health problems. PMID- 8717795 TI - Smoking among disadvantaged women: causes and cessation. AB - This study aimed to identify social-psychological factors associated with smoking and smoking cessation among disadvantaged women. Individual and group interviews were conducted with disadvantaged women in Atlantic Canada. Participants were predominantly poor, unemployed, geographically isolated, and single parents. The factors associated with smoking included coping with stresses, loneliness, powerlessness, low self-efficacy, social pressures, and addiction. Support from peers (i.e., women in similar circumstances) and partners was considered important. Health professionals and traditional cessation programs were not perceived as supportive. Participants viewed women's centres and women's agencies as appropriate deliverers of cessation programs. Methodological issues are discussed. PMID- 8717797 TI - Dilemmas of difference and the politics of comparison: cross-cultural research on dementia. AB - This paper examines some of the theoretical and methodological issues that were raised when I undertook cross-cultural research on dementia. The main concern addressed in this paper is the difficulty of representation within cultural groups. The paper contends that whilst representation can lead to perceived homogenization of a group, thus blurring intragroup differences, it is at times expedient to use this method in order to further the cause of that group. This paper proposes a way of presenting findings that, whilst meeting the aims of the research, compromises neither the participants nor the researcher. This paper also represents part of the decision trail the author left while carrying out the research. PMID- 8717798 TI - The politics of feminist ethnography. PMID- 8717799 TI - Knowledge, politics, culture, and gender: a discourse perspective. PMID- 8717800 TI - A struggle for equality: resistance to commissioning of male nurses in the Canadian military, 1952-1967. AB - Historical research was conducted to explore and describe the forces of resistance that prevented male registered nurses from being employed and conferred officer status in the Nursing Division of the Canadian military. Prior to 1967, only female nurses were permitted to join the Nursing Division. A 25 year struggle by the Registered Nurses Association of Ontario (RNAO), its Male Nurses Committee (MNC), and the Canadian Nurses Association (CNA) was required to change this discriminatory policy. The struggle for equality on behalf of Canadian male nurses was successfully resolved because of the united stand taken by the MNC, the RNAO, and the CNA. The study also demonstrates the need to move beyond matriarchal history perspectives in nursing to more completely understand the evolution of the profession in Canada. PMID- 8717802 TI - Long-term effects of a comprehensive rehabilitation programme after myocardial infarction. AB - The aim of this study was to determine the long-term effects on myocardial infarction (MI) patients of a six-month comprehensive rehabilitation programme (CRP) conducted by an interdisciplinary team regarding cardiac events, physical and psychological conditions, life habits, and cardiac health knowledge. The results of a multivariate analysis carried out five years after the MI showed that cardiac events and psychological condition were not significantly influenced by the CRP. However, it was found that the physical condition of the patients benefited from the CRP; self-reported physical fitness (p < 0.002) and physical exercise test (p < 0.007). CRP participation was linked to significant modifications of life habits (diet change; p < 0.04, sexual activity; p < 0.000). The cardiac health knowledge was significantly improved by participation in the CRP (basic cardiac knowledge; p < 0.005; knowledge about misconceptions; p < 0.04). In conclusion, CRPs have had positive long-term effects on physical condition, life habits and cardiac health knowledge. No such effects, however, were found regarding either cardiac events or psychological condition. PMID- 8717801 TI - [Predictors of regular physical activity in participants in a Canadian health promotion program]. AB - A series of predisposing and facilitating factors were examined to determine their contribution to the adoption of regular physical activity. The original longitudinal study Healthstyles was a comparative, before and after design, in which the participants (N = 640) were involved for 18 months in a multibehaviour health promotion program. The theoretical framework of Green, Kreuter, Deeps, and Partridge (1980) used for this study, the PRECEDE Model, reveals the role of three categories of factors at different levels of influence. In a correlational study, the sedentary subjects (N = 377) of a secondary analysis of data, were classified at the end of the program according to two behavioral status: adoption or non-adoption of regular physical activity. The nature of the study, the results of the preliminary analysis and the testing of the hypothesis, required a multivariate analysis approach. The logistic regression technique was used to differentiate between the adopters and the non-adopters. The results regarding the role of the combined predisposing and facilitating factors, show that only some of the predisposing factors predicts the adoption of regular physical activity. The adopters are relatively younger (OR = 0.48) males (OR = 1.7) with good or excellent self-evaluated health (OR = 0.51) and with a high score at the onset of the program, on the internal locus of control scale (OR = 1.8). Moreover, they are not different from the non-adopters in their need for external support in the change process. The impact of these results has been examined in terms of target groups and of content and approaches for future health promotion programs and research. PMID- 8717803 TI - Assistive technologies in stroke rehabilitation from a user perspective. AB - The purpose of the study was to provide knowledge, from a user perspective, about the selection process for assistive devices. Thirty-eight persons who had suffered stroke were interviewed about their assistive devices, approximately one year after stroke. It was found that the patients had little influence on the choice of assistive devices and that the maintenance and follow-up routines varied among patients. In some cases follow-up was not done at all. Despite these deficits, the patients used their assistive devices as intended. PMID- 8717804 TI - Home care--a realistic alternative for bed-blocking stroke victims in acute hospital wards? The conceptions of caregivers in six disciplines concerned. AB - Up to half of the stroke patients admitted to acute hospital wards become bed blockers. Investigations have been carried out in an effort to identify factors related to this problem. Very little is known about options which may lead to an alleviation of this problem. We investigated to what extent, in the opinions of professional representatives of six disciplines, home care can contribute to a solution. Sixty-nine stroke patients who were actually blocking beds in an acute hospital ward were described and examined on paper by a multidisciplinary panel. These patients were all moderately to severely disabled and needed a high degree of help in activities of daily life (ADL) and household activities. Estimations of the number of patients who were judged to be suitable for home care varied, although there was a fair degree of agreement between panel members concerning those patients who could and those who certainly could not return to their homes. Concerning one-third of the patients, the opinions of the caregivers diverged. Factors relating to the judgement of each panel member are identified. A method for selecting patients to be substituted to lower levels of care is suggested and discussed. PMID- 8717805 TI - Patients with chronic renal failure and their ability to cope. AB - The subject studied in this paper, describing uremic patients' use of coping strategies and their sense of coherence, was 48 consecutively selected patients with chronic renal failure at a Swedish hospital. The questionnaires used were the Jalowiec Coping Scale (JCS-40) and the Sense of Coherence Scale (SOC). The results showed that the patients used a variety of coping strategies, such as maintaining control, finding out more about the situation and hoping. In the use of coping styles, the confrontational style was the most favoured one, compared with the palliative and the emotive styles. There was a significant distinction between the predialysis patients and those on dialysis treatment concerning the use of strategies and coping styles. Patients on dialysis treatment showed significantly higher scores in overall strategies and in confrontational and palliative coping styles. The results revealed significant correlations between the JCS and the SOC scores, age and duration of kidney disease. Knowledge of the patients' commonly used coping strategies and sense of coherence is important when nursing care is being planned for the patient. PMID- 8717806 TI - Worse functional status among old people when admitted for arthroplasty--an evaluation with a new assessment system. AB - The aim of this study was to investigate whether there is a difference in functional status between younger and elderly people referred to an orthopaedic clinic for total joint replacement. A total of 709 patients with osteoarthritis of the hip or knee (aged 27-91 years) and 42 healthy controls (aged 42-83 years) were examined with a new Functional Assessment System (FAS) for lower extremity dysfunction. Age effects were explored by one-way analysis of variance. Older people showed higher dysfunction scores in almost all variables, with the exception of pain, where there was an inverse relationship, i.e, old people had lower disability scores. This age-related increase in disability scores was not observed in the control group. Old people seem to be referred for joint replacement on different grounds from those of younger people, despite the fact that age was not included in the criteria for arthroplasty. The results may indicate a hidden, age-related criterion in the selection of patients for arthroplasty. Different interpretations are discussed. PMID- 8717807 TI - Violation of the patient's integrity, seen by staff in long-term care. AB - The aim of the study was to describe the views held by staff (n = 233) in long term care on what they regard as a violation of patient's integrity and to define the concept "integrity" in simple terms. The design of the investigation was inductive. The answers were coded into 775 items. Agreement was reached between the authors on 770 of the items when sorted into 13 categories: the unique personality, autonomous self, personal opinions, secret self, personal competence, professional self, family self, cultural self, information self, personal properties, private territory, corporal self, and ridiculing. The number of categories shows a great variation in the ordinary language definitions given by staff. It was also shown in two other ways: the difference in the level of abstraction in the items, and the fact that integrity referred to the patient, directly or indirectly. Our conclusion is that integrity by the staff reflects an ethical value and not a personal trait. If the staff have to violate the patient's integrity, because other vital values are involved, it is absolutely essential that they consider the ethical aspects of the situation carefully. Such violation demands that they preserve their respect for the patient to the greatest extent possible. It seems important in the training of different staff groups to show the complexity in the concept "integrity" and the ethical dilemmas that arise, especially in the interaction with vulnerable patients who are not able to protect and maintain their own integrity. PMID- 8717808 TI - Living with cancer: perceptions of well-being. AB - The aims of the study were, first, to provide descriptive data on the perception of psychological well-being one year after surgery in a group of patients treated for gastro-intestinal cancer; secondly, to compare aspects of psychological well being between the group of patients living with other chronic diseases, and thirdly, to determine whether the perceptions of psychological well-being were related to coping capabilities. Sixty-nine patients with gastro-intestinal cancer and 20 patients with ostomies due to Crohn's disease or ulcerative colitis participated in the study. The most interesting finding in this study was that the cancer patients with a strong sense of coherence perceived their well-being as better than did the patients with a weaker sense of coherence. Other interesting findings were the differences in psychological well-being between younger (< 67 years) and older (> or = 67 years) cancer patients and between the younger cancer patients and non-cancer patients with ostomies. The young perceived their well-being as poorer than that of the old and the cancer patients rated their sense of psychological well-being as poorer than that of the patients with ostomies. It is concluded that nurses should focus their attention not only on the patients' physical well-being but also on their psychological well-being. Specific consideration should be given to the psychological needs of younger cancer patients. PMID- 8717809 TI - Some characteristics of the quality of life in old age illustrated by means of Allardt's concept. AB - This study illustrates some characteristics of the quality of life in old age when ageing is seen as a continued human development. In the Kungsholmen Project 87 healthy elderly persons were asked about the quality of their lives. Allardt's definition of quality of life was used as the conceptual framework for the content analysis. The findings indicate that the dimensions of loving and being, as Allardt describes them, take on a different meaning and the material things in the having dimension become less important. The differences point to another meaning of the quality of life in old age. The emphasis is on health and independence, contentment and a peaceful life, personal integrity in terms of a moral and a caring attitude. The findings tend to be in accordance with the successful ageing as described in Erikson's theory of psychosocial development and in Tornstam's theory of gerotranscendence. To gain a deeper understanding of this complex phenomenon, qualitative methods which go beyond definitions are required. PMID- 8717810 TI - How beds are used at a University Hospital--a study of bed and bedside activities. AB - Hospital beds have to fulfil many demands, and the features of the bed will consequently affect the comfort of both patients and staff at the ward. In the present study the use of hospital beds at Sahlgrenska University Hospital in Goteborg was investigated. The study was performed in two somatic wards; one general surgery ward and one internal medicine. The exact time of the observations as well as the categories of usage and activities performed on and at the bedside by the caring staff and the patients were registered. The study shows that there is a substantial difference between the two wards. In the surgery ward, the beds to a great extent play an active part in medical therapy and treatment, whereas beds in the medicine ward are used for vegetative recovery such as sleep, rest and alimentation. The personnel spent only a few hours with the patients, while they were in their beds, most of the caring occurring away from the bed. The results of the present study concerning how beds are utilized in modern hospital care should be considered when purchasing and designing hospital beds. PMID- 8717811 TI - Could department of health policy help to control methicillin-resistant Staphylococcus aureus? PMID- 8717812 TI - The recreational drug user in the intensive care unit: a review. AB - This paper describes the mode of action of a set of recreational drugs that may cause patients problems that are severe enough to warrant admission to an intensive care unit. The mechanism of harm will be examined as well as some of the strategies used to deal with these problems. A general view of the chosen group will be outlined, followed by a breakdown of the drugs into specific categories, with review of the harmful effects of individual drugs. It is the intention of the authors to review depressant drugs at a later stage. PMID- 8717813 TI - An exploratory study of patients' memory recall of their stay in an adult intensive therapy unit. AB - This study carried forward an exploration to determine if there is a need to visit patients in a ward following discharge from an adult intensive therapy unit (ITU), by ascertaining what patients remember of their stay and their ability to cope with this experience. Data were collected during interviews 48 hours following subjects' discharge from the ITU, and collated using an adaptation of thematic content analysis as described by Burnard (1991). Twenty-six subjects were studied over a 6-month period. The findings suggest that patients have vivid recollections of their stay in an ITU, and are consistent with previous research in respect of patients continuing to experience dreams, pain, sleep deprivation and worries about transfer out of an ITU to a ward. New themes suggest that ITU patients do not recall their "named nurse", but do recall detailed explanations given to them by nurses in the ITU. Patients also believe they would benefit from a follow-up visit to discuss and clarify aspects of their stay which were unclear or are causing them concern. PMID- 8717814 TI - A cognitive model for assessing depression and providing nursing interventions in cardiac intensive care. AB - Depression is an emotion experienced by each individual as some point in his or her lifetime. For some, the feelings are temporary, such as when one feels momentarily let down. For others, the feelings are deeper and may last for longer periods of time. Deeper and longer lasting depression may occur when individuals are confronted by certain unfavourable types of life situations such as a major physiological loss. For a once healthy individual, the depression associated with a physiological loss has penetrated through the individual's prior coping process and defences. Depression is a particularly common problem in individuals with a medical illness (Cavanaugh 1983). Heart disease is often experienced as a major loss for patients. Reports of depression in patients with coronary artery disease have ranged from 18% to 60% (Clark 1990). A broad range of physical disorders are commonly associated with depression. Among the most prominent disorders is congestive heart failure (Buckwater & Babich 1990). For depressed congestive heart failure (CHF) patients, a critical care nurse needs to assess the factors contributing to depression and recognise behaviours reflective of depression to be able to make appropriate nursing diagnoses and devise a plan to manage the patient's depression. To help critical care nurses accomplish this goal, this article contains an examination of depression as it applies to CHF patients according to Beck's cognitive triad (Ulerman et al 1984). PMID- 8717815 TI - Understanding balloon mitral valvuloplasty: the Inoue technique. AB - Severe mitral stenosis has traditionally been treated with surgical splitting or replacement of the mitral valve. A less invasive option is available, in which the value is stretched by an inflated balloon inserted via the femoral vein (percutaneous balloon mitral valvuloplasty). The most widely used method of mitral valvuloplasty, the inoue technique, is described in detail. Pertinent aspects of patient management and nursing care are also described. PMID- 8717816 TI - Children in intensive care: can nurse-parent partnership enable the child and family to cope more effectively? AB - The admission of a child to an intensive care unit (ICU) causes extreme distress and is likely to throw the family unit into turmoil (Hazinski 1992). The effects of hospitalisation on the child are well known (Rutter 1981), as are stressors which affect the parents' functional ability at a time of extreme stress (Rushton 1990). This paper examines the conflict caused for the whole family by a child's admission to ICU and how nurses can minimise these effects. In particular, it includes investigation of the use of a nurse-parent partnership which, it is argued, will enable the child and family to cope more effectively with the experience of intensive care admission. PMID- 8717817 TI - Haemostasis and disseminated intravascular coagulation. PMID- 8717818 TI - The needs of relative visiting adult critical care units as perceived by relatives and nurses. Part I. AB - This was a descriptive study, aimed at identifying the needs of relatives while they were visiting adult critical care units. A convenience sample of 24 critical care units participated, involving 351 critical care nurses and 255 visiting relatives. Using an adaptation of Molter's Critical Care Family Needs Inventory, the objectives of the study were: To compare (in ranking order) relatives' perceptions of how important their needs were with how important critical care nurses perceived them to be. To identify how satisfied relatives were with how their needs were met, while visiting the unit. To compare who relatives perceived to be the most appropriate person to fulfill each of their needs, with who nurses assessed to be the most appropriate person to fulfill the relatives needs. Relatives highlighted the importance of re-assurance and their need for sufficient information about their family member. Critical care nurses were only moderately accurate in assessing how important relatives' needs were, according to the findings from this relatives' group. Relatives were 'satisfied' with how the majority of their needs had been met. Only 10 of the 30 needs were considered to have been met to 'very satisfactory' level by at least 60% of the sample group. Nurses were identified by relatives as being the most appropriate person to fulfil 19 out of the 30 needs. Whereas nurses perceived themselves to be the most appropriate people to fulfill 25 of the 30 needs for relatives. The implications for clinical practice are also discussed. PMID- 8717819 TI - Intensive care syndrome: a literature review. AB - The aim in this literature review is to describe the definitions, denominations, clinical signs and symptoms, explanations, causative factors and interrelationships of the intensive care syndrome discussed since 1950. It was found that there is no agreement about which symptoms should be included in the syndrome, when the syndrome may appear and how many patients may be affected. Furthermore, it is unclear what causes the development of the syndrome; most authors conclude that there are many reasons for it. The syndrome has generally been examined by using a medical or psychological approach, but during the last few years it has also been described and analysed from a nursing care perspective. From this nursing perspective the syndrome may be seen as an individual pattern developed by patients during their stay in an intensive care unit (ICU) and sometimes this pattern of clinical signs and symptoms lasts for a shorter or longer period even after discharge from the unit. From a wider viewpoint the development of the syndrome can be seen as an increase by degrees or as a vicious circle. finally, most authors agree that the ICU syndrome consists of, and is caused by, a complex interaction between many factors. PMID- 8717820 TI - Intrahospital transportation: monitoring and risks. AB - Patients' intrahospital transport is considered as part of the mediconursing care continuum, since patients frequently need diagnostic or therapeutic procedures not performed at the bedside (Waddell 1975, Rutherford & Fisher 1986). Severe complications, such as airway obstruction, artificial airway or intravenous line removal, arterial blood gas and blood pressure changes, and arrhythmias or cardiac arrest, have been described during this transportation, especially in critically ill patients (Taylor et al 1970). Risks can be diminished if the patients are appropriately selected and carefully monitored during transportation (Venkataraman & Orr 1992). PMID- 8717821 TI - Non-steroidal anti-inflammatory analgesic injections. AB - The non-steroidal anti-inflammatory drugs (NSAIDs) are an important group widely prescribed in general medical practice. the availability of a few NSAIDs in injectable form provides an important approach to pain management. PMID- 8717822 TI - Informed refusal and patient autonomy: using reflection to examine how nursing knowledge and theory affect attitudes. PMID- 8717823 TI - Physicians' assistants: legal implications of the extended role. PMID- 8717824 TI - Care and management of peripherally inserted central catheters. AB - It is estimated that 85% of all patients admitted to hospitals in the USA in 1991 were recipients of a vascular access device (Barbone, 1995). Down-sizing of hospitals and the shift towards more outpatient and day-care procedures has meant that patients admitted to hospitals in the UK are more likely to be recipients of a vascular access device than they were 10 years ago, as their conditions are more likely to be acute. This article will discuss how, with careful management, peripherally inserted central catheters can improve the reliability and quality of vascular access for many patients. PMID- 8717825 TI - Living with eczema: the dermatology patient. AB - This article gives an overview of eczema, including its assessment, the needs of patients with eczema and the management of eczema. The prevalence of all forms of eczema is unknown but there is evidence that atopic eczema is increasing. Nurses can play a vital role in the management of this condition by giving patients time for explanation and discussion. PMID- 8717826 TI - Interpreting problematic behaviour in people with learning disabilities: 2. AB - The main theme of the first part (Vol 5(8): 546-50) of this two-part article was a focus on understanding the causative factors of problematic behaviour in people with a learning disability. A logical sequence to the identification of causes of problematic behaviours is that interactions to lessen such behaviours are identified. In this article, a discussion on interventions to guide nursing practice is offered. A short list of some interventions which emanate from the identified causes is presented in Table 1 for guidance; these interventions are analogous with many nursing and related theories (Peplau, 1952; Johnson, 1980; Wolfensberger, 1983; Roper et al, 1985; Jenner, 1992; Burford, 1993; Quinn and Mathieson, 1993). It is suggested that there is need for a paradigm shift to take place in the interpretation and understanding of problematic behaviours, and it is further suggested that nursing theories should be utilized within such a paradigm. PMID- 8717827 TI - Nurses remain critical to the success of community-based care. PMID- 8717828 TI - The public image of the mentally ill and community care. AB - Transferring the care of mentally ill patients to the community is not a new initiative. The first comprehensive Hospital Plan by the Department of Health and Social Security (DHSS, 1962) recognized and recommended that areas of health care which did not need to take place in hospital should be carried out in the community. Some 30 years later, this transition is taking place. This transition has not been a smooth process and in certain cases it has failed. The public image of mental illness has played a significant role in delaying the transfer of care to the community, although this is not the sole hindering factor. Poor liaison between health and social services, closure of hospitals before development of community facilities, inadequate community support and resources, and sensationalized media coverage have all been cited. Community care of mentally ill patients is a reality and it has been demonstrated that it can be well planned and properly funded (Faugier, 1993). Rather than dwelling on the shortcomings surrounding the implementation of community care, the way forward is to learn from mistakes and to identify means of increasing public acceptance of people suffering from mental illness. PMID- 8717829 TI - Discharge of psychiatric patients into the community: how many more must die? AB - Since the advent of community care, mental health services are increasingly coming under scrutiny, particularly the management of people with mental health problems in the community. Several murders committed by psychiatric patients in the community have fuelled public debate. This article reports on a study examining the recommendations of the Department of Health regarding the discharge of psychiatric patients from hospital to community care. It describes the background to events such as the Clunis report and guidance from the Department of Health on the management of mental health services. Semi-structured interviews, questionnaires and document analysis were used to ascertain the views of community psychiatric nurses and managers regarding the discharge of patients into the community. The findings identify areas of good practice and the need for nurses to improve their knowledge of the care-programming approach, risk assessment and inter-agency collaboration. PMID- 8717830 TI - Continuing care needs for the elderly mentally ill. AB - The demand for continuing care of the elderly, mentally ill continues to rise. The Government's latest response to this increase is to issue new guidelines on the responsibilities of the NHS in meeting continuing health-care needs. The general principles underpinning the guidelines are welcomed; however, this article identifies a number of problems with them regarding the elderly mentally ill. PMID- 8717831 TI - Eye examination and vision testing. AB - This article discusses some of the more common eye problems that nurses may encounter during their day-to-day practice. It describes some of the observations and simple testing that may be used to help determine the need for specialist consultation. A principal aim is to remind nurses that by the use of careful observation and simple examination techniques, they can play an important role in the detection of ocular pathology and the unmet need for specialist services. Manifestations of a variety of disorders are highlighted, together with some of the key points for nurses to consider when examining the external eye and lid. PMID- 8717832 TI - Decision making in professional practice. AB - There is a large body of work relating to the nursing process but comparatively little on decision making. Two main views exist, namely the rationalist and the phenomenological perspectives. It can be argued that the rationalist perspective is a quantitative approach whereas the phenomenological perspective offers a more holistic view. Intuitive judgment presents problems as it is only used by experts and is difficult to teach. In professional practice the decision-making process and the nursing process work together. PMID- 8717833 TI - Is preceptorship the answer to reality shock? PMID- 8717834 TI - Developing a political consciousness for children. PMID- 8717835 TI - Establishing therapeutic boundaries as patient advocates. AB - Pediatric nurses often struggle to define their advocacy role with patients and their families. Nursing is based on the therapeutic use of self and caring. An unclear therapeutic relationship can undermine nurses' advocacy efforts. Nurses are encouraged but often not guided about staying within the parameters that define a professional and therapeutic relationship between the nurses and the patient/family. Examining the impact of unclear boundaries on the patient/family relationship and the nurse's ability to serve as an advocate provides the basis for suggested constructive strategies for addressing the related issues. PMID- 8717836 TI - Exploring the boundaries of pediatric practice: nurse stories related to relationships. AB - Pediatric nurses struggle to find the right level of involvement with children and families. The purpose of this study was to illuminate nurses' struggles and insights as they learned to find the interpersonal boundaries of their own practice. The phenomenological method of Heideggerian hermeneutics was used to analyze data from audiotaped and transcribed single interviews of five pediatric nurses. The analysis of the transcriptions involved multiple stages of interpretation using a research team. The themes that emerged from the data were: (a) pediatric nurse as family caregiver; (b) finding the right level of care: over-involvement vs. crossing the line; (c) caring and the dying child; and (d) caring for the caregiver. Implications for pediatric nursing practice are explored. PMID- 8717837 TI - Therapeutic relations decision making: the Rainbow Framework. AB - The giving and receiving of gifts, funeral attendance, post-discharge babysitting, usurping of parental roles, indiscriminate sharing of personal information, and other issues prompted a pediatric tertiary care facility in the Midwest to examine intricacies involved in the promotion and maintenance of therapeutic relationships. A multidisciplinary group developed a therapeutic relations decision-making framework to (a) foster a proactive process of conscious deliberation regarding nurse/patient interactions, and (b) afford a nonthreatening mechanism for retrospective review of apparently non-therapeutic relationships. The framework, based upon an "act utilitarian" approach to moral reasoning, promotes the clarification of personal philosophy and values. The framework provides a basis for deliberate, consistent, and confident nursing care delivery and the impetus for developing of new patient and family care initiatives. PMID- 8717838 TI - Childhood drowning: review of the literature and clinical implications. AB - In children the risk of submersion is highest in those under 5 years of age and in male adolescents 15-19 years of age. This review examines the research literature that identifies factors related to drownings and near drownings, strategies to prevent these occurrences, and suggestions for primary prevention and associated nursing interventions. PMID- 8717839 TI - Mastery motivation and temperament of 7-month-old infants. AB - PURPOSE: To examine relationships among infant mastery motivation, temperament, and cognition with the goal of highlighting infant behaviors to which nurses should be alert in the clinical environment to promote optimal infant development. SAMPLE: Subjects were 26 healthy, full-term infants, age 7 months, recruited from a well baby clinic. METHOD: A descriptive correlational design was used. Tools included the Bayley Scales of Infant Development, Fagan Test of Infant Intelligence, the Dimensions of Mastery Questionnaire, and the Revised Infant Temperament Questionnaire. RESULTS: Cognition was not related to either mastery motivation or temperament. Several relations between mastery motivation and temperament emerged. Infants with high mastery pleasure were rated as more cooperative and less difficult, and tended to be rated as more active and less irritable. Independent mastery, on the other hand, showed no correlation with temperament scores. Infants who were rated as high in persistence on the mastery motivation questionnaire were rated as more cooperative, more rhythmical, and less difficult on the temperament questionnaire, and they tended to be rated as more approachable and less irritable. Infants who were rated as high in competence tended to be rated as less difficult. CONCLUSIONS: While temperament and mastery motivation are related, information on mastery motivation provides additional information to the nurse that may be helpful both in planning interventions to promote infant development and in providing anticipatory guidance for parents. PMID- 8717840 TI - Policy to limit young people's access to the media. PMID- 8717841 TI - Children with HIV becoming adolescents: caring for long-term survivors. AB - As children infected with HIV live longer, pediatric nurses who care for these patients need to be familiar with adolescent development and to anticipate physiologic and behavioral changes that will occur in this population. Concerns regarding confidentiality, autonomy, medication needs and compliance may become paramount. Learning how to interview adolescents and becoming comfortable with adolescent sexuality issues will enhance the patient-provider relationship. PMID- 8717842 TI - Ritual circumcision: educating parents. AB - Ritual circumcision, an ancient tradition with deep religious significance, is practiced by many cultures including those of the Jewish faith. As with any surgical procedure, there are some risks involved. Parents who choose to have their child circumcised in a ritual manner would benefit from the teaching that an informed nurse could provide in order to make an informed choice. PMID- 8717844 TI - Ethical issues surrounding attention deficit hyperactivity disorder. PMID- 8717843 TI - You pierced what? AB - Adolescents have discovered body piercing. Specific information about the piercing procedure, the sites, and the risks are presented along with an educational information sheet that can be distributed to pediatric clients and their families. Further nursing involvement in the community is stressed with one goal being the development of a collaborative education program for body piercers and their counterparts, tattoo artists. PMID- 8717845 TI - Fathers: the forgotten parent. AB - Fathers today face a confusing set of values and role choices. The challenges are particularly great for fathers of children with chronic illness or disability. Fathers must reexamine priorities, come to terms with losses, and develop and strengthen caretaking capabilities. The isolation commonly felt by fathers of children with special needs places the entire family at risk. Conversely, paternal involvement has many benefits. Professionals can take many steps to engage men in becoming supportive and important figures in the lives of their children. PMID- 8717846 TI - Central line associated bacteremia in the pediatric patient. AB - PURPOSE: To define the per-day risk of central line associated bacteremia in an infant-toddler population and to describe risk factors associated with the development of central line bacteremia. METHOD: The Central Line Data Tool collected information on 102 central venous catheters from 73 patients ranging in age from 1 day to 29 months. Each line was in place for 3 days or longer. FINDINGS: There were 17 documented catheter-related infections during the 1-year study period (7.7 infections per 1,000 catheter days). Factors significantly associated with central line bactermia included: PAS infusion, catheter type and site, medication administration, blood withdrawal, and accidental line disruption. CONCLUSIONS: Use of central lines for multiple purposes should be minimized. PMID- 8717847 TI - Personalizing cardiovascular terminology: measuring preload, contractility, and afterload. AB - For many nurses who are the heart (and soul) of institutions, the terms preload, contractility, and afterload are abstract terms learned in school but quickly forgotten. Yet, professional nursing effectiveness, like cardiac function, is ruled by these concepts, and it is important to relate these to the work day in order to gain a better understanding of this vital terminology. PMID- 8717848 TI - Pediatric management problems. What is your assessment? Secondary amenorrhea. PMID- 8717849 TI - Cardiac toxicity with cisapride. AB - Serious cardiac arrhythmias including ventricular tachycardia, ventricular fibrillation, torsades de pointes, and Q-T prolongation have been reported in patients taking cisapride with other drugs that inhibit the cytochrome P450-3A4 hepatic enzyme. This column will review the mechanism of action of cisapride, the drug's metabolism and reported cardiac adverse effects with intravenous cisapride. PMID- 8717850 TI - [Informed consent]. PMID- 8717851 TI - [Certain aspects of informed consent]. PMID- 8717852 TI - [Hospitalizations by force. Considerations regarding the law of June 27, 1990]. PMID- 8717853 TI - [Informed consent in psychoanalysis]. PMID- 8717855 TI - [Psychosis. Care contract and institutional treatment]. PMID- 8717854 TI - [Children and psychotherapy]. PMID- 8717856 TI - [Consent by suggestion]. PMID- 8717857 TI - [A different kind of care. Emergence of a request for care, psychologic disorder and social complaint]. PMID- 8717858 TI - [The schizophrenic patient]. PMID- 8717859 TI - [The double inversion or consent to experimentation]. PMID- 8717860 TI - [Importance of interdisciplinary relations: a new experience: an ethics council in psychiatry]. PMID- 8717861 TI - [A disoriented and jovial woman]. PMID- 8717862 TI - [Decubitus ulcer. Many measures help--but we have not yet conquered it]. PMID- 8717863 TI - [Emergencies in the hospital with special references to resuscitation in the hospital]. PMID- 8717864 TI - [The importance of back training in the therapy of osteoporosis]. PMID- 8717865 TI - [Standardized prevention of decubitus ulcers using the nursing process model. A sample of learning targets for the practical teaching in an intensive care unit]. PMID- 8717866 TI - [Decubitus documentation in the operating room]. PMID- 8717867 TI - [Testing the efficacy of a material for the relief of pressure in the prevention and treatment of decubitus ulcers]. PMID- 8717868 TI - [Surgical therapy of decubitus ulcers]. PMID- 8717869 TI - [Use of pulsed electrical stimulation in the therapy of decubitus ulcers]. PMID- 8717870 TI - [Mattress protectors from an hygienic and an economical point of view]. PMID- 8717871 TI - [Dying: surrounded by life. Living to the end--dying as a part of life"]. PMID- 8717872 TI - 'Nursing ethics into the next millennium'. PMID- 8717873 TI - Community nurses and health promotion: ethical and political perspectives. AB - This paper brings together ideas from two perspectives on ethics and health promotion. A discussion of the ethical dimension of the health promotion practice of community nurses is set in the wider context of health policy, with particular reference to health gain and individual responsibility. It is widely held that nurses have a key role to play in health promotion and that this is particularly the case for nurses working in primary health care. This assumption is reinforced by policy documents from the World Health Organization, the Department of Health and statutory bodies such as the United Kingdom Central Council for Nursing, Midwifery and Health Visiting. The approach of many nurses to health promotion has tended on the one hand to be somewhat naive and on the other to be authoritarian and didactic; there has been little discussion in the nursing literature of the ethical aspects of health promotion. However, recent developments in nurse education, such as Project 2000 and the consequent changes to preregistration programmes, have resulted in increased attention to both ethics and health promotion within the curriculum. PMID- 8717874 TI - Provider choice: essential to autonomy or advertising gimmick? AB - Free choice of provider is heralded as a right of autonomy, but the goals of autonomy are better served in today's health care environment when there is informed choice of the care delivery system. The principle of liberty is distinguished from respect for autonomy. Free choice of provider would be demanded only by liberty, except that allocation of health care resources does not meet criteria for the application of liberty. Patients attempting to choose the best practitioner do not have data to support the decision. If data were available, access to these superior practitioners would become an allocation problem. A mythology of the wise practitioner fosters the concept of provider choice as a personal judgement about clinical knowledge. The emerging trends of collaborative care, standardization of practice guidelines and diversity of delivery systems among reimbursers create a situation where a patient's autonomy to choose goals for treatment is respected through the choice of delivery system. PMID- 8717875 TI - Respect for autonomy: is it always paramount? AB - Following the argument proposed by Tschudin in 1986 that many nurses do not have the skills for ethical decision-making, this article identifies and discusses one ethical problem from practice. The problem concerns an extremely obese patient who refuses to be moved by a hoist. The nurse acquiesce to the patient's wishes and she is moved manually by four members of staff. The issues identified for discussion are: the paramountcy of the principle of respect for the patient's autonomy; the rights and obligations of the nurses; and the justification for influencing patient choice. The approach used by the ward nurses is analysed and the value of using an ethical decision-making model is considered. PMID- 8717876 TI - Client age, gender, behaviour: effects on quality of predicted self-reactions and colleague reactions. AB - This comparative study shows biases relative to client age, gender and behaviour demonstrated by 268 female nurse subjects. A repeated measures design was utilized. All three main effects were significant (p < 0.001) for how respondents predicted that they would react to various clients and also how they predicted that their colleagues would react. Most two-way and three-way interaction effects were significant. Subjects demonstrated more favourable reactions to nice, young, male clients and least favourable reactions to not nice, old male clients. Study subjects predicted more favourable self-reactions to simulated clients than for their colleagues. What should be important to every professional as a result of this study is the need to be aware of self-biases, which may cause errors in decision-making and nursing care interventions and lead to ethical violations with clients. PMID- 8717877 TI - Ethical dilemmas faced by the Nursing Disciplinary Committee. AB - This article presents the complicated ethical dilemmas that arise in the procedures and proceedings of the Nursing Disciplinary Committee, which deals with matters of life and death (e.g. patients' rights to quality and safe care, professional integrity and accountability, and the nurse's future). The article also describes the composition and function of the Committee, the type of case it deliberates, its limitations, and the dilemmas its members may encounter. PMID- 8717878 TI - To kill or not to kill: a question of wartime ethics. AB - In this article, the author describes ethical decision-making in unique circumstances. A dichotomy exists between the dual roles of nurse and disaster manager in a wartime setting. The circumstances of the situation had never been faced before and no precedents existed for the type of decisions being made. Clearly, professional codes of conduct existed along with international conventions with reference to war. The circumstances required the author to challenge the concepts of teleology and deontology in a search for the most fitting answers to a unique problem. His aim was to try to create the greatest good out of an impossible situation. The author reflects on his actions in the light of ethical thinking and considers whether his decisions were right. PMID- 8717879 TI - Ethics in the intensive care unit: a need for research. AB - Intensive care units are challenging and technologically advanced environments. Dealing with situations that have an ethical dimension is an intrinsic part of working in such a milieu. When a moral dilemma emerges, it can cause anxiety and unease for all staff involved with it. Theoretical and abstract papers reveal that having to confront situations of ethical difficulty is a contributory factor to levels of poor morale and burnout among critical care staff. Despite this, there is a surprising dearth of published nursing research in the UK that investigates how staff deal with ethical issues in intensive care units. The purpose of this paper is to explore and discuss the development of a research framework designed to explore how staff deal with moral dilemmas in a British intensive care unit. PMID- 8717881 TI - Suicide and voluntary active euthanasia: why the difference in attitude?: a response. PMID- 8717882 TI - Row over professional boundaries simmers on. PMID- 8717880 TI - The right to touch and be touched. AB - Touching is an integral part of human behaviour; from the moment of birth until they die, people need to be touched and to touch others. Touching is an intimate action that implies an invasion of the individual's personal, private space. In ethical terms, the question of touching is closely related to the patient's right to integrity and inviolability. The purpose of this study was to describe touching as it is experienced by elderly patients and nurses in long-term care. Touching was approached as a form of communication and as an important part of nursing practice. The participants, 25 patients and 30 nurses, were interviewed using a semistructured schedule. The data were analysed using the method of content analysis. The patients experienced touching by nurses as gentle, comforting and important. The nurses, for their part, experienced touching by patients as easy and natural. The patients rarely touched nurses more than was necessary. In some cases, nurses had to cope with violent touching by patients. Some women nurses interpreted touches by male patients as having a sexual nature and as annoying. This had taught male patients to avoid touching nurses. On the other hand, friendly and grateful touches by patients were very important to nurses. When used for emotional purposes only, touching presupposed a good relationship between nurses and their patients. Although touching is extremely common in nursing practice, there has been very little research into its meaning. More work is therefore needed to explore the role and meaning of touching in nursing. PMID- 8717883 TI - Taskforce to review paediatric ITU training. PMID- 8717884 TI - Huge profits from NHS data bank. PMID- 8717885 TI - Decision time. PMID- 8717886 TI - Universities can do it better. PMID- 8717887 TI - Public health: water pressure. PMID- 8717888 TI - A shift in time. PMID- 8717889 TI - Problem peanuts. PMID- 8717890 TI - Standard setting: a joint venture. PMID- 8717891 TI - Accessing library services. PMID- 8717892 TI - The psychosocial implications of back injury at work. AB - The majority of studies on back injury have been quantitative in nature, focusing on ergonomic or financial implications of back injury problems for trusts and the profession. The psychological and social effects upon the individual suffering the injury have largely been ignored. This qualitative study sought to redress the balance by examining why injuries were occurring. It focuses on the personal cost of back injury to nurses and its effect on their lives and careers. Questionnaires were sent to members of the Work Injured Nurses' Group (WING) which is funded by the Royal College of Nursing. The findings revealed that a large proportion of accidents were preventable. Nurses who sustained a back injury suffered both physically and psychologically, and generally felt unsupported and isolated. This, together with a lack of redeployment opportunities, seemed to affect adversely their ability to adapt to their injury and remain in the profession. PMID- 8717893 TI - Discharge planning: the need for effective communication. AB - This article emphasises the need for effective interdisciplinary and interagency communication to ensure a satisfactory transfer of care between hospital and the community. The author highlights some of the policies and guidelines which inform current practice, and examines some of the reasons why care is often fragmented in this important area for patients and their relatives. PMID- 8717894 TI - Complementary therapies and research in nursing practice. PMID- 8717895 TI - Contraception and the under-16s: the legal issues. AB - This article discusses the legal issues and the implications of consent related to providing contraceptive services within general practice for girls under the age of 16. Possible scenarios and interactions are also discussed to allow the reader to apply principles to practice. PMID- 8717896 TI - Hysterectomy (continuing education credit). PMID- 8717897 TI - Nurse writes "HIV+" on lab order: test without consent. PMID- 8717898 TI - Bloody fetus handed to mother for photo: emotional distress. Case in point: Brown v. Philadelphia College 674 A. 2d 1130--PA (1996). PMID- 8717900 TI - Court orders board to license nurse under "reciprocity". Case in point: Murry v. Dept. of Prof. Regulation 664 N.E. 2d 716--IL (1996). PMID- 8717899 TI - Legal case briefs for nurses. NY: student nurses blow whistle: nurse terminated for abusing patients; IN: Board reverses ex parte suspension: untimely appeal-- "no finding" stands. PMID- 8717902 TI - Mixed mode education learning and the use of textual resources. AB - Economic and market forces, as well as beliefs about sound educational practice, are prompting faculty to adopt mixed mode education, drawing upon both established and in-house material production. There is much to be gained, both from the student, purchaser and provider (institution and tutor) point of view, but implementation of such programmes requires careful planning. This paper stems from the 1994 Nurse Education Tomorrow Conference (Durham) and discusses pragmatic considerations when textual resources are used to fuel education innovation. It argues that the use of 'study guides' may free precious tutor resources for higher taxonomy roles, and afford opportunities to review the ways in which the management of effective learning can assist in the development of quality assurance criteria. PMID- 8717901 TI - On refereeing journal articles. PMID- 8717903 TI - Why teach sociology? A contribution to the debate. AB - This paper seeks to engage in the debate about the role of sociology in nurse education that has been conducted in the pages of Nurse Education Today by Hannah Cooke (1993) and Keith Sharp (1995). Cooke's paper is addressed first. It is argued that in her application to nursing of Foucauldian notions of power, Cooke inflates the salience of power to the development of holistic care. In doing do, she diverts her argument away from the central problematic of her paper, namely her identification of critical structural approaches as being essential to nursing sociology. Cooke's conclusions on this issue are defended against the attack made upon them by Sharp. First, it is contended that Sharp's argument that the multiparadigmatic nature of sociology means that it cannot act as a guide for nurses is self-contradictory. Second, it is argued that his characterization of nursing work as essentially instrumental is misguided, in that the prosecution of holistic care entails a more reflexive form of action. Finally, it is posited that his interpretation of Cooke's perspective as narrow political vision is unsustainable. PMID- 8717904 TI - Assessment of clinical and communication skills: operationalizing Benner's model. AB - The introduction of ward-based clinical assessment of nursing skills in the 1970s meant a more realistic method of assessment, but still essentially a 'snapshot' of the student's practical abilities. This now being replaced by continuous clinical assessment which requires that student nurses meet the expected level of competence in a range of nursing skills in each clinical placement. Benner's (1984) model of skill acquisition is currently receiving considerable attention by nurse educationalists and is providing the framework for many curricula (English 1993). The model identifies five stages of development in nursing: novice; advanced beginner; competent; proficient; and expert. What is needed, however, is a more detailed description of what those stages mean in terms of the component parts of nursing, especially the clinical and communication skills. This paper describes a framework for the assessment of clinical and communication skills, a framework which seeks to operationalize Benner's model by defining the level of performance expected in clinical and communication skills at each stage. PMID- 8717905 TI - Training to do women's work in a man's world. AB - Many student nurses also carry the responsibilities of parenthood, and government has encouraged the recruitment of more mature students to nursing. The majority of nursing recruits continue to be women, but career models are more appropriate to those who do not bear the principal responsibility for childcare, namely men. After reviewing the literature relating to these issues in the National Health Service, the article discusses discrimination against nurses who are parents. An empirical study of a group of mature nursing students on Project 2000 courses is reported, showing that they encounter problems with childcare, support, study, family relationships and inflexibility within nursing education. The article concludes by suggesting a number of points which nurse educators should bear in mind when planning and delivering courses to ensure that the needs of student parents are met. PMID- 8717906 TI - More semi than structured? Some problems with qualitative research methods. AB - Qualitative research techniques have become a favoured tool of academic nursing research in recent years. Although there is now a significant body of methodological literature intended to train students in these techniques, the limits of qualitative research techniques are rarely discussed. This research note discusses the limits on claims than can be made about the utility of such techniques. PMID- 8717907 TI - The nurse teacher as Action Researcher. AB - It is important that the work of nurse education and nurse educators is critically evaluated. Just as it is no longer acceptable for clinical practitioners of nursing to engage in nursing activities out of a sense of tradition or a history of habit and practice, so too must nurse educators engage in systematic evaluation of their own craft. A number of authors have described Action Research's suitability for nursing (Webb 1990, Greenwood 1984, MacGuire 1991). In this paper the authors examine Action Research from the perspective of nurse teachers and suggest that it is an appropriate method for studying nurse education. PMID- 8717908 TI - Learning contracts: an Australian experience. AB - This paper explores the use of learning contracts - a tool that can facilitate self- directed learning that is specific to an individual student's needs. Reflection on the experience of using contracts within an Australian university highlighted some perceived benefits for learning and teaching as well as potential limitations and areas of concern. Recommendations for effective use of contracts in teaching nursing include: that teachers should be adequately prepared for additional roles required in the implementation of contracts; the tool itself should be simplified and easy to read; and, students should be assisted to set realistic goals and to think about quality learning. PMID- 8717909 TI - CD-ROM: a piece of technology or a reliable 'friend'? AB - The reactions of students new to CD-ROM databases were evaluated through a combination of observation and interviews. There was much similarity between the observed behaviour of the new users and that demonstrated by individuals in the development of interpersonal relationships. It is further argued that the users' reactions to the CD-ROM hardware and software parallel the levels of absorption in aesthetic experience and cannot be explained in purely behavioural terms. The friendliness of CD-ROM database applications has implications for the development of information technology competencies in new users. PMID- 8717910 TI - Quality in nurse education: an exploration of the concept of students as customers. AB - The issues of Quality and Quality Assurance in nurse education, have become prominent in the design, management and marketing of courses. However, quality is a concept which has many meanings. One of the central ideas which has emerged from the current discourses on quality, is that of the 'customer'. In the new business jargon of the 1990s, with its emphasis on the market and competition, the importance of 'satisfying the needs of the customer/consumer' has been a dominant theme. One of the customers of nurse education is seen as the student. The reconceptualization of the student as customer may carry with it, connotations which do not readily fit into the context of education and learning as a jointly negotiated process. A collaborative, rather than consumerist model is proposed as a preferable basis for the ensuring of quality, and the involvement of students in that process. Such a model, would be in line with a truly student-centred approach to nurse education, where power is shared between those involved in the process. The importance of the current inequality of power in student-teacher relationships is a barrier to such an approach and true student participation, rather than tokenism is called for. PMID- 8717911 TI - Educational issues in preparing community health nurses to use nursing diagnosis with population groups. AB - Recently there has been increased interest in the use of nursing diagnosis by community health nurses who work with population groups in community settings. The purpose of this article is to discuss educational issues important in the preparation of undergraduate students and practising community health nurses in the use of nursing diagnosis with population groups. The educational issues discussed emerged from the findings of a preliminary survey of undergraduate students and community health nurses and were related to: differences in the learning requirements of novice and expert practitioners; common errors in the use of nursing diagnosis; and the perceived benefits and barriers in using nursing diagnosis. There is a need to develop an educational strategy to address these concerns for both undergraduate students and community health nurses. PMID- 8717912 TI - Reconceptualizing the theory-practice gap in mental health nursing. AB - The purpose of this paper is to identify the key problems in bridging the gap between theory and practice in the education of mental health nurses and to propose a strategy for overcoming these problems. The paper begins by reviewing the existing literature on the subject of the theory-practice gap, and identifying the unique problems which mental health nursing faces in attempting to address them. It then goes on to propose a unified model of clinical nursing practice, nurse education and user involvement, which would place nurse educators at the forefront of radical reform of mental health services. PMID- 8717913 TI - [Change in nursing paradigms. 2. Elizabeth Barrett's "Theory of Power"]. AB - The second part of this article deals with the fundamentals of Martha Rogers' Conceptual System, as developed by Elizabeth A. M. Barrett in "Theory of Power". Some concepts from the work of Margaret Newman are discussed, examples are used which are taken from Rogerian science based nursing practice. PMID- 8717914 TI - [Searching for the biological roots of self-care and care behavior. 1. The evolutional importance of self-care and care]. AB - In search of biological roots for self-care and for caring behavior. Part 1: Evolution and the significance of self-care and of care. Care and self-care are not exclusively human activities. They occur widely in the animal kingdom, in the form of hygiene related and pro-social behaviour. This has fundamental evolutionary significance. Examples of caring among animals will be used to point out the existence of biological roots for human caring behaviour. Self-care and care can, theoretically, be classified within the concept of self-preservation, largely congruent with nursing theories and models and with relevant research. The possibility therefore exists to develop interdisciplinary research programmes with a view to gaining biological insights about the origins and the meaning of caring behaviour. This will help to understand that the human body is a medium for expression and interaction in the context of nursing. PMID- 8717915 TI - [The importance of trust in nursing]. AB - This qualitative study is based on a dissertation which was submitted as part requirement towards the Higher Diploma, Level II, "Clinical Nursing Expertise". Observation revealed that trust must be regarded as a prerequisite for all nursing interventions. 20 patients were the subjects of open-ended interviews. They were asked what, for them, was the meaning of "trust", and what determined whether they could trust the nurses. Analysis of the data showed that emerging categories depended on different factors. Though technical competence and organisational structure were not unimportant, personal competence of the nurse was crucial. The patient expects a helpful relationship, which presupposes a high level of social and communication skill. In order to be able to come up to the expectations which patients have of the nurses' personality, appropriate training and supervision is essential. PMID- 8717916 TI - [Humanistic care--a vital dialog with myself as a nurse]. AB - The author documents the use of "humanistic care" as described by Paterson & Zderad (1988) and presents two case studies. Humanistic care is a transactional process, the carer entering into a relationship with the person for whom he/she cares. At the same time the carer embarks on a conscious dialogue with him/her self. To be "present" means being conscious of ones own thoughts, feelings, impressions and reactions. During a course of graduate study in Canada, the author enrolled in a "self-directed learning programme". She set herself the task of self-analysis. She conducted interviews with two patients suffering from cancer and with their relatives. She hoped that she would be able to reexamine, in this new situation, feelings and emotions which she had repressed earlier in her hospital nursing experience. The goal of the learning experience was to develop personal "authenticity" by becoming aware of existential experiences (her own and others'). PMID- 8717917 TI - [Approach to foreigners--the contribution of ethnology to nursing]. AB - The starting point of this analysis is the concept of "The stranger", as discussed in "transcultural nursing" in the USA and currently researched by cultural anthropologist. The aim is to designate common areas of interest between nursing sciences and cultural anthropology and between ethnological and nursing practice. It is pointed out that more space is needed in nursing theory and practice, for the discussion of the concept of "stranger" and that this should not be confined to the "exotic ethnic" stranger. PMID- 8717918 TI - [As we make our bed, so we must lie. The effect of super soft mattresses on the mobility of the elderly needing care]. AB - This article discusses the effect of supersoft mattresses on the mobility of elderly persons in nursing homes. It is based on a pilot study, which also includes video footage of patients engaged in a variety of activities after having slept on soft mattresses for an extended period of time. The soft mattresses were then replaced by conventional hospital mattresses, and the same activities were videotaped again. The individual motion sequences were analyzed and compared. The results of this pilot study can be summarized in the following hypotheses: soft mattresses 1. restrict mobility; 2. cause reduced orientation; 3. impair human function, particularly perception. These findings are highly significant because reduced mobility is often the decisive cause of a decubitus hazard. Supersoft bedding can increase this motion deficit. While the pressure relieving and decubitus-preventing effects of soft mattresses are undisputed, the study confirms that motion aspects and their impact on the overall condition of the patient must be considered in an appraisal of the nursing situation. Nursing personnel must have access to specific criteria governing the use of soft bedding material. They need to develop nursing skills and abilities that make supersoft bedding superfluous or that limit the primary and secondary effects of such measures. PMID- 8717920 TI - [Professional pedagogy in nursing--considerations on graduate education of nursing teachers 2]. AB - The concept previously presented determines the educational process, showing it to be related to what is happening on the ground. Starting from there, this part develops a basic course structure which is limited in its direction and reflects broadly what education is about. PMID- 8717919 TI - [Innovative processes in ambulatory care--AIDS as a pilot project]. AB - Flexibility in nursing practice and the willingness to be innovative are discussed, using the care of patients with AIDS as an example. When this new infectious disorder made its first appearance many countries tried to provide a patient-centred, community based service, in spite of the severity of the illness and the fact that it was usually lethal. A number of innovations were required to achieve this. The top priority was the provision of a community nursing service. This contribution consists of the analysis of the structure of provisions for care of patients with AIDS in the German Federal Republic (BRD). It deals with the reaction of the nursing services and asks to what extent the objectives have been achieved. Three different ways to adapt to the situation in the BRD are described and analysed. Data are based on qualitative interviews with members of the nursing service. It is shown that AIDS has led, in the BRD, to numerous innovations in community nursing care, the relevance of which far transcends the treatment of AIDS. In spite of this success, the care of patients is unsatisfactory. Many promising attempts to change came to grief as a result of resistance and obstacles to implementation. Others proved to be insufficiently robust or inappropriate in view of the multiplicity of problems encountered in AIDS. Reasons are to be found in the defective structure of the service, in the fact that management difficulties concerned with implementation of innovations were underestimated and that there is a "crisis" in nursing in the BRD. There have been criticisms for a long time, pointing out that the basic qualification is insufficient, that it does not prepare nurses adequately to deal with new social and health problems. If no change is introduced to training, community nursing is unlikely to be able to cope innovatively and flexibly with new challenges in the future. PMID- 8717921 TI - Reflecting on a nurse's personal experience with a stroke. AB - These reflections chronicle the experience of a retired nurse educator who had a stroke and shared her experience with the author. The format alternates between the first person remembrances of the nurse who had a stroke and the author's analysis of the nurse's experience. The article is followed by two nursing research perspectives. PMID- 8717922 TI - Cardiac rehabilitation in the elderly: improving effectiveness. AB - Heart disease is a major cause of mortality and morbidity among adults who are older than 65 years of age. One-third of cardiac operations on adults in the United States are performed on patients who are more than 65 years old, and almost half of the hospitalized candidates for an outpatient cardiac rehabilitation programs are in the older patient population. Structured educational program that include exercise and modification of risk factors have been shown to reduce the risk of subsequent coronary events; however, studies show that older adults enroll in these programs at a significantly lower rate than do patients in other age groups. Nurses and other healthcare professionals must eliminate barriers to participation and adapt their programs to meet the needs of older adults with cardiac disease to reduce morbidity, enhance functioning, and improve quality of life. In this article, the authors describe current knowledge about the efficacy and use of cardiac education and rehabilitation in elderly patients and suggest implications for future practice and research. PMID- 8717923 TI - A follow-up study of ethnic and gender differences in cardiac rehabilitation. AB - In a 1991 study, Romeo and Saccucci used the Sickness Impact Profile (SIP) to measure the perceived health status of individuals enrolled in a cardiac rehabilitation program and the perceived health status of those who received routine care. The participants were further classified by ethnic group and gender to enable the researchers to focus on the differential effects of cardiac rehabilitation on these groups. The present study was initiated to determine if the earlier results would remain consistent over time. The SIP was used to measure the health status of many of the same subjects within 3 years of the original measurement. The results of this study show fewer differences among the participants over the long term, which suggests that the time required for improvements may differ for various ethnic groups and for men and women. PMID- 8717924 TI - Cardiac rehabilitation survey: maintenance of lifestyle changes and perception of program value. AB - Graduates from a Phase II cardiac rehabilitation program were surveyed about their maintenance of healthy lifestyle changes from 1 to 4 years after completing the program. Patients were queried about their continued performance of routine aerobic exercise; use of safe exercise principles; adherence to a low-fat, low cholesterol diet; and surveillance of lipid levels. Results showed that 74% of patient graduates reported exercising aerobically three to four times weekly, and 50% used their pulse rate to guide exercise intensity. The patients were also asked their opinions about the value of the cardiac rehabilitation program; 91% of patients reported an improvement in their quality of life, and 97% felt the program was a good use of insurance dollars. The findings of this study have implications for program evaluation and modification. PMID- 8717925 TI - Beyond Type A: hostility and coronary heart disease--implications for research and practice. AB - Research into the psychological and behavioral correlates of coronary heart disease (CHD) has gone beyond the traditional description of the Type A behavior pattern and is focusing instead on components that may be more sensitive predictors of CHD endpoints. One component being investigated frequently is hostility. Many questions related to hostility and CHD remain unanswered and warrant further research. Implications for practice include a need for reexamination of standard risk reduction programs in light of current knowledge. PMID- 8717926 TI - Predictors of the use of memory improvement strategies by older adults. AB - This study examined the ability of depression, health, and memory self-efficacy scores to predict the use of internal and external memory strategies by cognitively impaired and cognitively intact older adults. There were no differences between the groups on memory self-efficacy scores and on the extent of their use of the external memory strategy of place. The cognitively intact group used strategies to aid memory more often than did the cognitively impaired group and used more external than internal memory strategies. The cognitively intact group used the internal strategies of elaboration and rehearsal more often than the other group and were less likely to expend effort than the other group. Memory self-efficacy scores were the sole predictor of use of external memory aids by the cognitively intact group, and age was the sole predictor of use of external memory aids by the cognitively impaired group. No variable was able to predict the use of internal memory strategies by either group. PMID- 8717927 TI - Rehabilitation nursing is "like a box of chocolates". PMID- 8717929 TI - Taking time. PMID- 8717928 TI - All in the cards. PMID- 8717930 TI - Genograms: application in hospice care. AB - Genograms are a comprehensive, systematic assessment tool that have valuable application within the hospice home care setting. They also can be used to assist the client and family in their journey through the dying process and the bereavement period. PMID- 8717931 TI - Medicare coverage of medical supplies. PMID- 8717932 TI - Mother as vocation: implications for home care nursing. AB - Because pediatrics has become a major component of home care practice, many children who are seen by home care nurses are visited in foster homes. This article discusses the dynamics of being a foster mother and outlines the ways the nurse can work with the child and foster family in implementing successful interventions. PMID- 8717933 TI - Morphine for Dolly. PMID- 8717935 TI - The Custom Nurse Program. PMID- 8717936 TI - Current management of hemochromatosis. AB - Community and home healthcare nurses can play important roles in identifying early symptoms of hemochromatosis, encouraging diagnostic screening and assisting with prompt treatment of this potentially fatal disorder. When hemochromatosis is detected early, organ damage often can be prevented. PMID- 8717937 TI - Admission nursing in home care: the exclusive domain of the rehabilitation nurse? PMID- 8717934 TI - Decreasing the prevalence of pressure ulcers using agency standards. AB - Home care patients either have or are at high risk for pressure ulcers. This article describes the outcome of a study that examined the effects of pressure ulcer prevention and treatment standards on the prevalence rate of a home healthcare agency's client caseload. Implications for practice and further research also are discussed. PMID- 8717938 TI - Nursing clinics in urban settings. AB - Decreased home care funding and the elimination of on-site clinics in elderly housing establishments by other organizations led to the creation of student- and faculty-staffed nursing clinics at Regis College. The initial two clinics, which have expanded to five in various sites, provide challenging clinical experiences for students, faculty members, and other members of the healthcare community. This article provides an overview of these clinics and examples of how they can work. PMID- 8717939 TI - Chart audit using the American Nurses Association standards of practice. AB - This article describes a study using chart audits as a strategy for continuous quality improvement within a home health agency's quality improvement program. Findings are discussed using the American Nurses Association Standards of Home Health Nursing Practice as a framework. PMID- 8717940 TI - Internet surfing for ease of access to AHCPR-clinical practice guidelines. PMID- 8717941 TI - Intraethnic diversity. PMID- 8717942 TI - Who will prepare nurses and others to work within the parameters imposed by health care reform policy? PMID- 8717943 TI - "Been there, done that!". PMID- 8717944 TI - Imagining the future for nursing administration and systems research. AB - Science-based nursing practice is shaped and influenced through the lens of research. These findings are dependent upon the phenomenon being studied and the interpretation that is given to the information discovered. In the emerging postmodern era, nursing, along with every other science-based discipline, must explore its work through the lens of quantum physics, chaos theory, and ecofeminist principles. This broader world view will enlarge our definition of health, nursing, and healing. An expanding perception of our world and our work will lead to relevant and life-giving models for care and caring that will powerfully position nursing as an essential player in the changing health care field. PMID- 8717945 TI - A theoretical model for nursing systems outcomes research. AB - Nursing research on patient and administrative outcomes has typically examined the relationships between selected structural characteristics and outcomes, without taking into account the organization's context. In contrast, health services research has focused on the relationships between the organization's context and outcomes, most often mortality, without taking into account structural characteristics. Although widely used, both approaches develop fragmented knowledge. This article describes a comprehensive theoretical model that takes into account the relationships among the organization's context, its structure, and both patient and nursing administrative outcomes. PMID- 8717946 TI - Evaluation research. AB - Evaluation is often described as different from research. In this article, common myths about evaluation are discussed as well as reasons these myths no longer fit with current understanding of evaluation research. Issues associated with the conduct of evaluation studies are described, with recommendations made for maximizing the rigor of designs. Suggestions are made for how nurse executives can shape and influence externally generated evaluation projects. PMID- 8717947 TI - Integrating research into acute care settings: reflections of two nurse researchers. AB - In this article, two clinical nurse researchers describe experiences with integrating a program of nursing research, including the use of the findings and methods of research and the conducting and disseminating of research, into two acute care hospitals. Strategic and contextual factors are specified in relation to targets within and outside of the hospitals. PMID- 8717948 TI - Research-based practice. AB - Research-based practice requires a practice setting that promotes the use of knowledge, access to current scientific knowledge, and the ability of the clinician to think critically about using the knowledge to positively affect patient outcomes. This article presents an overview of what is known about the research utilization process and describes a variety of approaches to facilitate research-based practice. PMID- 8717949 TI - Revisioning graduate education in nursing administration: preparation for a new paradigm. AB - Education for nurse administrators is at a crossroads. Current trends in health care administrative roles require a revisioning of the master's level educational preparation for future practice as well as to retain control of administrative education by graduate schools of nursing. A new approach is described that addresses the issues and challenges facing today's and tomorrow's administrator for patient care services. PMID- 8717950 TI - Joint appointments in practice positions. AB - Faculty practice has been a value in both nursing education and nursing service. Recent changes in education provide the opportunity for practice to be rewarded in the university setting. Changes in health care delivery demand expert practitioners and leadership in advanced practice nursing. The history of education-practice integration is reviewed. A new role of the practitioner educator (reverse joint appointee) is presented. Benefits of the position to both education and practice are discussed. PMID- 8717951 TI - The challenge for education in a transformed health care system. AB - A focus on health promotion, disease prevention, and health education for all members of society has emerged from the federal guidelines of Healthy People 2000 and, more important, from the demonstrated needs of vulnerable populations. New and potent organisms and diseases have stressed the health care system. Nursing administrators face significant challenges in managing health care delivery and nursing within the multidisciplinary team in the transformed system. In all areas, education must be directed to meet the challenges and nursing must assume the leadership for ensuring the health care needs of individuals and communities, now and in the future. PMID- 8717952 TI - [How we could save effectively...]. PMID- 8717953 TI - [The congress in words and pictures]. PMID- 8717954 TI - [18. Meeting of delegates of the Swiss Nursing Association. The Swiss Nursing Association is ready for new challenges]. PMID- 8717956 TI - [Mirjam's experiences in the hospital]. PMID- 8717955 TI - [The tricky placing of venous lines. The ideal venipuncture: short and painless]. PMID- 8717958 TI - [Disquieting results of a U.S. study on tendencies in medicine. Extension of life: often an extension of agonizing dying]. PMID- 8717957 TI - [Murderers on the ward]. PMID- 8717959 TI - [The health care system between economic rationale and medical ethics. False economic incentives in our system of health care]. PMID- 8717960 TI - [Anthropoligical approach to care. Reflections on the diversity of nursing]. PMID- 8717961 TI - [Fighting for less suffering]. PMID- 8717962 TI - [The economist's point of view: cascades of money for health]. PMID- 8717963 TI - Eric Caines is an unlikely ally to nurses. PMID- 8717964 TI - Government wants nurses to expand roles. PMID- 8717965 TI - Unpaid experience to be recognised by UKCC. PMID- 8717966 TI - Crisis point. PMID- 8717967 TI - Goodbye nurses. PMID- 8717968 TI - Inside view. PMID- 8717969 TI - They are watching me. PMID- 8717971 TI - Broken promises. PMID- 8717970 TI - Am I doing well? PMID- 8717973 TI - Doubly honoured. Interview by Charlotte Alderman. PMID- 8717972 TI - An aspirin a day. PMID- 8717974 TI - Educating teachers in children's illnesses: a study. AB - This study was carried out in response to teachers' concerns about emergency situations involving children with asthma, epilepsy and diabetes. Data were collected on whether teachers had received advice on coping with such emergencies in either their basic teacher training or in-service training, and whether they felt competent to cope with emergencies which could arise. The research found that no overall policy existed and that teachers would appreciate regular updates on the care of these children. The author also describes training sessions arranged following the analysis of the study. PMID- 8717975 TI - Nutritional status and chronic obstructive pulmonary disease. AB - This article explores the importance of an adequate diet for the health of the chronic respiratory patient. Poor nutrition can exacerbate or even cause respiratory symptoms. Therefore, both specialist and generalist nurses need to increase professional nutritional knowledge to address and co-ordinate nutritional aspects of care for patients and their carers. Next week, we will publish the results of a research project conducted by the author to examine the effect of specialist nurse intervention through nutritional education with these patients. PMID- 8717976 TI - The role of health psychology post-myocardial infarction. AB - This article examines the part health psychology has to play in underpinning care strategies and supporting professional practice during the process of rehabilitation following a myocardial infarction. PMID- 8717977 TI - The principles of pressure sore prevention. AB - This article describes the main causes of pressure sore development and discusses the principles of pressure sore prevention. It relates to UKCC Professional Development categories: Reducing risk and Care enhancement. PMID- 8717978 TI - [Stuttgart congress on "Alternative healing and caring methods. 1000 participants hope for gentle medicine]. PMID- 8717979 TI - [420 nurses at the symposium at the Munich University Hospital. That got under our skin]. PMID- 8717980 TI - [Oncologic nursing congress in Damp: new perspectives for hematologic-oncological patients]. PMID- 8717981 TI - [The hospital nursing schools are doomed to die]. PMID- 8717982 TI - [Nursing courses in Germany 3. Munich is oriented towards Christian humanity]. PMID- 8717983 TI - [Self-care for nurses. Tension pain--what to do?]. PMID- 8717984 TI - [Potato compress, as shown in the shoulder region]. PMID- 8717985 TI - [Alcohol in the work place. The hospital is a developing country]. PMID- 8717986 TI - [Definite work times and room service. Work satisfaction improves the length of stay of nurses in their profession]. PMID- 8717987 TI - [Relief from non-nursing duties: electronic data processing creates more time for the patients]. PMID- 8717988 TI - [A nursing concept is being realized into practice. 2. Only continuing education can assure adequate care]. PMID- 8717989 TI - [Care of incontinence in demented patients: talk to the patient in normal language!]. PMID- 8717990 TI - [Proposals for change in nursing insurance: new trickery has been discovered]. PMID- 8717991 TI - [Transfer of the elderly from the hospital to their home. Time structures in the home are self-determined]. PMID- 8717992 TI - Beyond the stages of sleep: an emerging nursing model of sleep phases. AB - The stages of rapid eye movement and non-rapid eye movement sleep are common topics in most nursing texts. It is impractical, however, to verify the stages of sleep in hospitalized patients. The article present a nursing model of sleep phases that uses observed behavior to identify which phase of sleep the adult patient is experiencing. The model is based on the findings from a qualitative study of 16 women hospitalized with myocardial infarction. Subsequent analysis of the data led to the development of Dreher's model of sleep phases. The retiring phase begins with declining arousal and preparations for sleep and continues until the onset of sleep. The resting phase begins with the onset of sleep and continues until the onset of final awakening. The rising phase commences with the onset of awakening and continues until the complete arousal. This model is presented as an emerging nursing model in which the behavior of sleep in hospitalized adults can be described. It is hoped that the model can be used to help nurses conceptualize sleep pattern disturbances so that appropriate nursing interventions can be implemented. PMID- 8717993 TI - Sleep dreams of women in the childbearing years: a review of research. AB - Descriptions of dreams of women during pregnancy have appeared in the literature for more than a century. Daydreams and sleep dreams are differentiated. A review of the literature published during the last four decades uncovered a limited number of empirical articles. The general consensus is that the sleep dreams of pregnant women are indicative of their concerns regarding safety for self and for the expected child. The most recent information focuses on the use of groups to help pregnant women explore the content of their dreams. PMID- 8717994 TI - Sleep in pregnancy. AB - The article examines relationships between pregnancy and maternal sleep. Specifically, sleep as a restorative process is considered with regard for the metabolic and arousal demands of childbearing. The analysis draws attention to the limited number of studies in the area and the need for greater research interest in pregnancy sleep phenomena. The available evidence indicates that maternal slow-wave and rapid eye movement which are key to anabolic activity and neural-cerebral recharge, are protected throughout pregnancy until perhaps the last 3 to 4 weeks before delivery. The sleep disturbances about which term gravidas complain appear to be based on increased periods of wakefulness after sleep onset. PMID- 8717995 TI - Neonatal sleep patterns: implications for nursing. AB - The article describes newborn sleep patterns and includes a discussion of factors influencing neonatal sleep. An important nursing role is to teach parents about infant sleep and to reinforce American Academy of Pediatrics' recommendations on sleep position. Central nurseries, feeding schedules, and constant lighting affect newborn sleep behavior. Nurses can change policies and create an environment in the hospital that enhances parent-infant synchrony. Nurses also have a role both prenatally and in follow-up settings to educate parents about newborn sleep. PMID- 8717996 TI - Sleep in men and women infected with human immunodeficiency virus. AB - Little is known about sleep problems in persons with human immunodeficiency virus (HIV) infection. The article reports a study that assessed quantitative and qualitative aspects of sleep in a group of men and women with HIV infection and examined sleep parameters in regard to the degree of immune function. The convenience sample of 50 persons was drawn from an HIV clinic in a large midwestern metropolitan area. Subjects identified a variety of problems with their sleep that were not significantly related to their immune status. The findings of this study indicate that nurses should perform a sleep history and assessment on all persons with HIV infection to identify those in need of intervention. PMID- 8717997 TI - Symptom description and management in narcolepsy. AB - Narcolepsy is a neurologic condition that is chronic and lifelong. The study reported describes the experience of living with the most prevalent symptoms of narcolepsy and the management strategies employed for those symptoms by persons with narcolepsy. Descriptions of living with excessive daytime sleepiness, sleep attacks, and cataplexy are given. Nonpharmacologic strategies for symptom management were described by 63.2% of the study participants for cataplexy, 54.5% for sleep attacks, and 35.5% for excessive daytime sleepiness. Specific management and coping strategies are provided. PMID- 8717998 TI - Rest. AB - Rest has been examined only sporadically in the nursing literature, primarily as a physiological need closely related to sleep. The article describes the concept of rest from a variety of perspectives and lays the foundation for further analysis. Themes of physical, mental, and spiritual components of rest emerge as the concept is explored. Further analysis of the concept of rest in both health and illness is recommended. Exploration of the individual patient's perception of rest and restful activities seems to be a key to promoting and prescribing rest as a nursing intervention. PMID- 8717999 TI - Dreams: their function in health and illness. AB - Despite centuries of published theory and research, general agreement on the nature and function of dreaming continues to elude the scientific community. As theory and evidence accumulate and collide, the recounting and analysis of dreams continue as powerful clinical tools for healing self and patients. The article reviews major theories of dreaming, including a description of the basic structure of dreams and mental processes in dream formation; explores the usefulness of dreams in promoting health and dealing with illness through the analyses of three dreams; and identifies basic principles of dream exploration for use in clinical nursing practice. PMID- 8718000 TI - From the wings of night: dream work with people who have acquired immunodeficiency syndrome. AB - Nurses who work with seriously ill and dying patients are engaged in helping them live fully until they die. Most theories of death and dying deal with conscious activities only. Yet many people find relief form the fear and loneliness of life threatening illness if given the opportunity to express their totality, meaning their unconscious as well as conscious processes. Working with the mind may not be enough. Attending to the unconscious through dream work can allow inner healing to occur, which can support that person as he or she moves into the unknown. The article describes the AIDS Dreamwork Project, the goals of which are to understand patients better and to help them better understand themselves, to help them live and die more consciously, and to discern patterns in their dreams that may be unique to people with acquired immunodeficiency syndrome. Six themes have been identified in the dreams collected so far: wake-up dreams; reassurance dreams; dreams of opposition, separation, or coming together; transition dreams; transformation dreams; and spiritual or archetypal dreams. PMID- 8718001 TI - Eczema: making an unpleasant condition more bearable. PMID- 8718002 TI - Benign breast pain. PMID- 8718003 TI - Iron deficiency in eight-month-old babies. AB - Iron deficiency is the most commonly reported nutritional disorder of early childhood in the UK. Recently, blood samples taken from 1,075 normal 8-month-old babies in the ALSPAC study in Bristol showed that 23% were anaemic according to WHO criteria. Anaemia was not associated with poor socioeconomic circumstances. The more rapid the growth of the child, the more depleted the iron stores. The findings of the study suggest that the currently accepted normal values for iron levels in young children may be too high, but further research is needed. Infants whose main milk was cow's milk at eight months had the lowest intakes of iron and vitamin C. PMID- 8718004 TI - Remedies for common family ailments: 11. Relieving the pain of sports injuries. AB - Many sports injuries involve tissue damage which results in the release of chemical mediators such as prostaglandin. These stimulate the nerve endings, producing pain. Many topical analgesics, eg aspirin and NSAIDs, act by inhibiting these chemical mediators. Others work on the "counter-irritant" principle, or by producing local heat or cold. Anyone on other medication or with existing medical conditions should consult a pharmacist or doctor before embarking on analgesics. PMID- 8718005 TI - Current health advice for travellers and holidaymakers. AB - Nurse-led travel clinics are increasingly seen as an essential part of the service offered in many general practices. The emphasis is now on offering people a full travel health consultation rather than vaccination or antimalarial advice in isolation. In this way advice can be tailored to the individual's needs and informed consent can be given. Special factors apply to women who are pregnant or contemplating pregnancy, and the nurse or midwife needs to be able to give appropriate advice. Gastrointestinal upsets are a particular hazard for babies and young children travelling abroad and advice on food hygiene is important. PMID- 8718006 TI - Children with cancer: how parents view social work help. AB - The parents of 16 children with cancer discuss aspects of the social work help they received at the oncology unit of a children's hospital early in their child's illness. In a semi-structured interview schedule, parents identified the emotional trauma resulting from the illness as more significant to them than practical problems. Reactions to the prospect of referral to a social worker were negative, though actual contact proved helpful. Seeing a social worker may be more useful to parents if scheduled after they have had time to absorb information about the diagnosis and treatment plans. Results of the study made staff more aware of the psychosocial trauma suffered by parents, and of the need to explain to parents the help that the social worker can offer. PMID- 8718007 TI - Midwives and HVs: women need you after early miscarriage. PMID- 8718008 TI - Managing genetic information: implications for nursing practice. PMID- 8718009 TI - Pre-election speeches to healthcare workers. PMID- 8718010 TI - Pets for patients. PMID- 8718011 TI - Profit sharing partners. Interview by Kate Williams. PMID- 8718012 TI - Difficult birth. PMID- 8718013 TI - Zero tolerance. PMID- 8718014 TI - Get involved. PMID- 8718015 TI - Breaking down the barriers. PMID- 8718016 TI - Partners in care. PMID- 8718017 TI - The King's Fund network: sharing good practice. AB - In conjunction with the King's Fund, Nursing Standard will be publishing regular reports on developments in nursing around the UK. Each report will focus on a specific area of practice and provide details of units which have experience in each area. The information will be based on material taken from the King's Fund network database and network days. To coincide with the launch of free network membership and to start the series, this report describes how the Nursing Developments Network is run. PMID- 8718018 TI - Health education and COPD rehabilitation: a study. AB - Following last week's introduction to the subject of nutrition and rehabilitation of patients with chronic obstructive pulmonary disease (COPD), this article reports on a study of whether nurses can improve the diet of these patients when they are at home, by providing education about nutrition. The research demonstrated a positive correlation between dietary knowledge and understanding of the link between diet and health. It also showed that change was achieved as a result of education in these areas. Further research would be required to measure the effect of such changes on the long term health and quality of life for these patients. PMID- 8718020 TI - The present nurse education system is based on students' supernumerary status. PMID- 8718019 TI - Providing appropriate services for continence: an overview. AB - This article gives an overview of continence services, detailing the policies which should inform service provision. The author highlights the role of the specialist continence adviser and suggests relevant questions to be asked for the future development of a service which can be accessed by and meets the needs of all its users. PMID- 8718021 TI - High interest. PMID- 8718022 TI - Making our presence felt. PMID- 8718023 TI - Clear lessons to be learned. PMID- 8718024 TI - Telling friends. PMID- 8718026 TI - A problem shared... PMID- 8718025 TI - Professional development. Stroke: knowledge for practice (continuing education credit). PMID- 8718027 TI - Operating at the frontier of nursing. Interview by Jane Cassidy. PMID- 8718028 TI - Big issues. PMID- 8718029 TI - Elderly care counts. The view from the sharp end. PMID- 8718030 TI - Long-term prospects. PMID- 8718031 TI - Learning disabilities: the challenge for nursing. AB - Over the past 10 years, people with learning disabilities have moved out of institutions and into the community. Their care is now seen as a social, rather than a health, issue. But there is a danger that this may lead to their general health needs being overlooked. Specialist registered mental handicap nurses are ideally suited to bridging this gap, possibly in partnership with primary health care teams. PMID- 8718032 TI - The benefits of a reflective model of nursing. AB - The concept of reflective practice is proving popular among practising nurses. This paper describes a way of using reflection in practice. It also describes how reflective principles were used to underpin the development of a model to guide nurses' work. The difference between this approach and the traditional use of theoretical models is outlined and its value for guiding practice is discussed. A second article next month will look at the model in practice in more detail. PMID- 8718033 TI - Making sense of radiotherapy: delivery and safety. AB - This is the second of two articles looking at the administration of different kinds of radiotherapy. It examines what nurses should consider when delivering teletherapy, brachytherapy and unsealed sources of radiation. The issues involved in radiological protection are considered. PMID- 8718034 TI - The effects of chronic childhood illness on families. AB - Chronic illness in childhood has massive physical, social and psychological effects on families who, in addition to fulfilling their usual duties, roles and obligations, are expected to meet the needs arising from the child's illness. This constitutes a severe disruption to the ordinary continuity of family life. This paper and examines the effect of cystic fibrosis on families. It gives a brief overview of the physiological nature of cystic fibrosis, examines the effect of diagnosis on other members of the family and the process of adjustment they go through to accept the condition. Although CF has been chosen specifically, the effects on the family resemble those of most chronic conditions. PMID- 8718035 TI - A hell called the Somme. PMID- 8718036 TI - Complementary therapy. A taste of health. PMID- 8718037 TI - Old story, new ending. PMID- 8718038 TI - Infection control. Menace across the channel. PMID- 8718039 TI - Infection control. The battle continues. PMID- 8718040 TI - Health assessment of the older patient. AB - More than 50% of patients admitted to critical care units are older than age 60. Interpretation of assessment data should take into consideration the normal physiologic variations associated with the aging process. This article discusses age-related physiologic assessment findings of older persons pertinent to the critical care nurse. It also addresses some common physiologic aging problems older patients experience in the intensive care unit. PMID- 8718041 TI - Pharmacologic issues in aging. AB - Older persons represent the fastest growing segment of the population and are already known to be the most prominent consumers of medications in the health care industry. The factors posited to account for this phenomenon include the perception of physical disability and the increased incidence of chronic diseases in this population. The management of an acute illness in older patients is often complicated by adverse or unpredictable drug reactions. Nursing management of safe drug therapy in the critical care setting begins with an understanding of basic pharmacodynamic and pharmacokinetic concepts and how they are affected by both aging and critical illness. PMID- 8718042 TI - Nutrition support for the critically ill older patient. AB - Changing demographics will increase the geriatric population in the intensive care unit. Geriatric clients are more likely to be malnourished than their younger counterparts, making the early initiation of appropriate nutritional support vital. The nutrition support process adapted for critical care nurses is outlined. Strategies for identification and management of common complications are presented. PMID- 8718043 TI - Dysrhythmia formation in the older adult. AB - Cardiac dysrhythmia in the older patient is a common finding during critical illness. Changes in the functional anatomy and physiology of the cardiac conduction system are associated with the direct and indirect effects of aging. The pathophysiology and clinical manifestations related to changes in cardiac cellular and neural structures are presented. Knowledge of these changes will guide the critical care nurse in planning care aimed at producing positive patient outcomes. PMID- 8718044 TI - Age-related changes in renal function. AB - As the population in the United States ages, the number of individuals who will develop renal disease will increase for a variety of reasons. Some of the causes may be an acute problem such as acute renal failure. However, in most cases, renal involvement is a secondary problem that may be caused by age-related changes in the kidney along with therapy that is used to treat a primary disease process such as hypertension or diabetes mellitus. This article explains the age related changes that occur in the kidney and the clinical implications for treatment. Nursing care specific to the problems associated with age-related kidney changes is also discussed. PMID- 8718045 TI - Planning care for acutely confused critically ill older persons. AB - The critically ill older patient is at increased risk for developing an acute confusional state or delirium. Critical care nurses must be aware of the risk factors, the clinical manifestations, and potential complications associated with delirium. Nursing strategies focus on prevention, comprehensive assessments, and interventions to manage agitated behavior and provide environmental support. PMID- 8718046 TI - Optimizing ischemic stroke outcomes: an interdisciplinary approach to poststroke rehabilitation in acute care. AB - Stroke is the third leading cause of death and the leading producer of disability among adults in the United States. Despite the high incidence of stroke, the general public's understanding of stroke symptoms, risk factors, and the need for early assessment and treatment is poor contributing to delays in treatment and the potential for suboptimal outcomes. Ischemic stroke accounts for most stroke events. Treatment is expanding to include drug agents that lyse clots and medications that may interrupt the neuronal metabolic cascade occurring with ischemia. Rehabilitative strategies require synergistic, collaborative relations among providers to optimize outcomes. PMID- 8718047 TI - Alcohol abuse in older persons: implications for critical care. AB - Alcoholism in the older population is a significant, yet often hidden or unrecognized problem. With the growing older population, a concurrent increase in the number of geriatric patients admitted to critical care settings will occur. An understanding of alcoholism in older persons, coupled with an early recognition of the problem by the critical care nurse, is vital in minimizing the consequences of alcoholism and withdrawal during critical illness. Prompt and appropriate treatment of manifestations of withdrawal is necessary to improve patient outcomes for the elderly individual suffering from alcohol abuse. PMID- 8718048 TI - Special issues in critical care gerontology. AB - The numbers as well as the acuity level of the geriatric critical care patient are increasing and will continue to do so well into the 21st century. There are certain characteristics and special issues or considerations that are unique to the geriatric population. It is therefore imperative that the critical care nurse know and have an appreciation for the uniqueness of the geriatric patient to provide quality care and promote the best possible patient outcome. This article addresses clinically relevant aspects of geriatric critical care patients and offers suggestions as to how their care should be managed. PMID- 8718050 TI - The quality of life improvement model: a means to guide practice. AB - Guiding primary health care with quality of life (QOL) in mind is essential to the survival of a practice as well as the health care system. The present system's aggressive methods to increase quantity of life must be abandoned in order to focus on QOL. With the advent of managed care systems, with or without national health care reform, the health care focus will be less on high technology and more on high-quality assessment, prevention, and health promotion. Promoting and following a practice model that advocates QOL and choices for care should be an integral element for guiding primary health care in the present and future. Utilization of the QOL improvement model will allow the clinician to assist varying clients to achieve enhanced QOL. PMID- 8718049 TI - Violence and the older adult: the role of the acute care nurse practitioner. AB - The United States population has an increasing number of older adults. One health problem often unrecognized in this aging population is elder abuse. Elder abuse can be classified into six categories: (1) physical abuse, (2) sexual abuse, (3) financial exploitation, (4) neglect, (5) psychological abuse, and (6) violation of rights. The abused older adult often lives with the abuser, has no outside support, and may live with intergenerational family violence. The acute care nurse practitioner is in an excellent position to become a patient advocate by recognizing the abused older adult, initiating an interdisciplinary team approach, and educating the community about elder abuse. PMID- 8718051 TI - Virginia Henderson's contribution to nursing. PMID- 8718052 TI - Using vignettes to collect data for nursing research studies: how valid are the findings? AB - Vignettes are simulations of real events which can be used in research studies to elicit subject's knowledge, attitudes or opinions according to how they state they would behave in the hypothetical situation depicted. Advantages associated with the use of vignettes as research tools include: the ability to collect information simultaneously from large numbers of subjects, to manipulate a number of variables at once in a manner that would not be possible in observation studies, absence of observer effect and avoidance of the ethical dilemmas commonly encountered during observation. Difficulties include problems establishing reliability and validity, especially external validity. This paper considers the advantages and disadvantages associated with the use of vignettes as data collection tools, concluding with a check-list to help critique vignettes studies. PMID- 8718053 TI - Striving to prevent falls in an acute care setting--action to enhance quality. AB - Although most falls do not result in serious physical injury, they can contribute to a loss of confidence and mobility which can culminate in a significant reduction in quality of life. Furthermore, the potential to fall is often increased when an individual is institutionalized because of frailty or confusion. The purpose of the study was, therefore, to establish whether a structured intervention would assist in preventing falls in an acute setting. This pre-test/post-test study was carried out over a 12-month period. Interventions included risk assessment, an alert system, reinforcing preventive actions, staff education and ongoing audits and feedback. Initial analysis of the data and comparison of fall rates indicated a significant reduction in the rate of falls between the pre- and post-intervention phases, although subsequent statistical analysis did not identify any significant relationships. It must be noted that no controls existed for extraneous variables, although patient profiles varied minimally during the period of the study. Outcomes include: a reduction in fall numbers and rates, enhanced staff morale with ownership of the programme, provision of a learning experience for staff (on which to build), and the fostering of a professional approach to improving the quality of patient care. PMID- 8718054 TI - Community mental handicap nursing services in Northern Ireland: a profile of clients and selected working practices. AB - The aim of this survey of community mental handicap nursing in Northern Ireland (NI) was, among other things, to construct a profile of clients of the service as well as to describe selected working practices. A 25-item questionnaire was developed and administered to all 50 community nurses working with people with learning disabilities in NI. The number of responses was 36, giving a rate of 72%. Fifty-six percent of the clients were female. Young to middle-aged adults made up the largest portion of the caseload. Over 80% of the clients had no history of hospital admission. Caseload size also differed according to the years of service, gender and grade of the nurses. The main reason for visiting clients as reported by respondents was to give physical care. PMID- 8718055 TI - Self-medication in acute surgical wards. AB - Self-medication of oral drugs is not current practice on surgical wards. We have introduced a system for suitable patients in which they give to themselves all their oral medication while in hospital. This was introduced in stages to gain the confidence of staff. Many patients wish to self-medicate; this system allows better use of the drugs that they bring into hospital from home; more appropriate timing when in hospital and speed up discharge procedures, as the patient is already familiar with, and has a supply of, their drugs to take home. PMID- 8718056 TI - The decision to prolong life: ethical perspectives of a clinical dilemma. AB - As a result of rapid developments in medical technology, health-care workers are increasingly faced with decisions regarding the prolongation of life and the withdrawal of treatment. Such decisions are often extremely complex and present a moral dilemma: they are further complicated by prevailing politico-economic, social and cultural influences. This article contrasts deontological and utilitarian philosophical perspectives, and discusses ethical principles which impinge on the decision making process. A knowledge and understanding of these philosophies and principles are essential to assist clinical practitioners in the examination of moral issues surrounding life and death. PMID- 8718057 TI - Health, health promotion and the elderly. AB - Health promotion is a mode of practice which is being increasingly examined by policymakers (DHSS, 1987; DoH, 1992). Although practitioners are being required to screen people over 75 years of age and are exhorted to reduce accidents in the elderly by 33% between 1990 and 2005 (DoH, 1992), there is evidence that they do not value this sort of work (Pursey & Luker, 1993). This paper looks at the findings from 178 interviews with people aged over 75 years, examining the importance of health and health promotion to the elderly. Further to the work of Cox et al. (1987), it is clear that people aged over 75 years continue to engage in a wide variety of activities which are designed to keep or improve their health. Elderly people's accounts of their health suggest that the real influence of social circumstances and environment must be assessed and planned for if health promotion activity is to have relevance and meaning to this group of people. Evidence is presented which indicates that the elderly are a group of people who would welcome health-promotion activity provided it is given in easily accessible forms. The notion of client participation is highlighted as a difficult area, which is likely to require particular skill in working with elderly clients. In the light of these findings, practitioners may need to examine their own attitudes to their work with the elderly. PMID- 8718058 TI - Audit or research--what is the difference? AB - Audit and research are activities which have some characteristics in common and others which are rather different. This paper presents working definitions of each process and then examines their attributes and interrelationships in some detail. Areas covered include their purposes, what types of process they are, their theoretical bases, the methods used in each, sampling, the use of findings, confidentiality and the time frame of each. If nurses are to use or undertake research and/or audit appropriately in their practice, it is important that the similarities and differences between the two are clear. PMID- 8718059 TI - An evaluation of staff training in psychological techniques for the management of patient aggression. AB - Training for staff working with violent patients is frequently recommended, sometimes implemented, but rarely objectively evaluated. In this longitudinal study 47 nurses attending a training day to learn strategies for coping with violent psychiatric patients were compared with 108 non-attending control group subjects. The rate of assaults on staff on wards taking part in the study was 31% lower after implementation of the training and wards sending a majority of staff to the training experienced a particularly significant reduction in assaults. The effectiveness of this 1-day training package is discussed. PMID- 8718060 TI - Creating consensus about nursing outcomes. II. Nursing outcomes as agreed by patients, nurses and other health professionals. AB - The questions 'what is it that you want from nursing?' and 'how would you know when you have got what you want from nursing?' were put to patients, nurses and other health professionals. The patients identified essential qualities of nurses. Problems within the nursing group are identified. The view of each group on the other's work is discussed. Areas of consensus related to respect, communication, competence, co-operation and context. Indicators or measures of the consensus areas are reported. PMID- 8718061 TI - Hong Kong nurses' health-related behaviours (HRB). PMID- 8718062 TI - [Prof. Kunze: "No therapy without attention to smoking behavior"]. PMID- 8718063 TI - [Hygiene in intensive care medicine 3]. PMID- 8718064 TI - [Prevention of burnout and addiction]. PMID- 8718066 TI - [Efficiency orientation versus quality of care]. PMID- 8718065 TI - [Who should pay for this ...]. PMID- 8718067 TI - [The ranking of nursing diagnosis in the nursing process]. PMID- 8718068 TI - [The tasks of the ward nurse and the positive effects on the patient. Copy of a paper in the field of special education for administrative personnel]. PMID- 8718069 TI - [Risk of infection in endoscopy. Measures for the protection of nursing personnel]. PMID- 8718070 TI - [Murau-Stolzalpe nursing school in Amsterdam]. PMID- 8718071 TI - Stroke. The role of the nurse. PMID- 8718072 TI - Problem that cuts both ways. PMID- 8718073 TI - Two-edged sword of publicity. PMID- 8718074 TI - Pomp and certain stances. Interview by Ian McMillan. PMID- 8718075 TI - Job swap. PMID- 8718076 TI - Double bind. PMID- 8718077 TI - Muddy waters. PMID- 8718078 TI - Hormone replacement therapy and its role in the menopause. AB - The physical and psychological effects accompanying the menopause can be distressing. Some 75-80% of all women develop at least some symptoms due to the sudden withdrawal of oestrogen. This paper reviews the use of hormone replacement therapy which can alleviate most menopausal symptoms. PMID- 8718079 TI - The development of leadership skills in clinical practice. AB - Leadership is often seen as the province of the manager. However, as the NHS changes and responsibility is increasingly devolved, practitioners at all levels are expected to lead change and innovation. In this paper the author describes how attending the Leadership in Nursing for the Community (LINC) programme has influenced her practice. As a result of the programme she has an increased awareness of her own leadership role and of the political and professional climate in which she works. PMID- 8718080 TI - How to design an effective research study. AB - The previous paper in this series (March 13) examined three methodological issues: sample selection, treatment and/or intervention and observation and measurement. Here, aspects of research design and procedures will be considered. Research design is important in helping to ensure that the results can be correctly interpreted and that observed changes can be attributed to the planned intervention rather than to other intervening factors. The description of the procedure explains how the selected methods were combined and describes the practical details of how the study was carried out. PMID- 8718081 TI - Systems of life. The pancreas. PMID- 8718082 TI - Living under the shadow of illness. PMID- 8718084 TI - Failing to make the grade. PMID- 8718083 TI - Absolutely flabulous. PMID- 8718086 TI - Priced out of practice. PMID- 8718085 TI - Room for improvement. PMID- 8718088 TI - Craniosacral therapy. PMID- 8718087 TI - Vitamin K: friend or foe. PMID- 8718090 TI - Positioning nursing within a reformed health care system. PMID- 8718089 TI - The registered nurse surgical assistant. AB - This article provides a candid account of the evolution of two Registered Nurse Surgical Assistant positions, and their roles as members of a large open-heart team. A description of the expectation of the job, the role in surgery and the challenges and rewards are outlined. PMID- 8718091 TI - Shared visions & hallucinations: acquiring a taste for chaos. PMID- 8718092 TI - Latex allergy update. PMID- 8718093 TI - Reclaiming the power to heal. PMID- 8718094 TI - Protection for patients or providers? AB - So-called patient advocates are taking many political action steps supporting greater public control of the managed care industry. Managed care organizations contend that they are being bashed unfairly, often by provider groups trying to safeguard their economic interests under the mantle of protecting patients. With billions of dollars and thousands of jobs at stake, the battles and spin doctoring are unlikely to subside in the near future. PMID- 8718095 TI - Risk shifting in subacute care. AB - The implications of risk shifting on subacute care providers by payers are enormous. Financially, it will force subacute care providers to strategize on how to deal with capitated arrangements. Strategically, it will provide the impetus for retaining, increasing or losing market share in the health care arena. Philosophically, it will forge a leveling of the playing field among different subacute care sites in terms of average length of stay and pricing. PMID- 8718096 TI - Back to basics--Part I: System selection. AB - Technology management involves continually making decisions about upgrades, changes or adjustments. Suggestions for managing the process of system selection and ongoing technology management are given. PMID- 8718097 TI - Long-term care update. AB - While the legal cases against health care providers continue to increase, the cases against acute care facilities are declining as the number of cases against long-term care facilities and employees rise. Nurses need to be aware of the differences in legal issues in the extended care delivery system. PMID- 8718098 TI - Quality wound care equals cost-effective wound care: a clinical model. AB - The cost of wound care is often defined as the cost of materials and labor, ignoring the effect of both variables on outcome. As a result, money-saving measures may actually increase cost while reducing the quality of care. Definitions and tools to enable professionals to understand, evaluate and compare the cost-effectiveness of wound care practices and treatment modalities are given. PMID- 8718099 TI - Managing material resources in the ED. AB - A negative variance in materials resource in one hospital's emergency department revealed that the unit was losing money in medical-surgical supplies. The Roy Adaptation Model was applied to assess the problem in the present system. PMID- 8718100 TI - Successful interview strategies. AB - Every newly hired staff member will affect hundreds of patient outcomes as well as the success of the team at large. An interview determines the applicant's values and identifies if he/she is congruent to the professional practice environment. PMID- 8718101 TI - Computer system failure: planning disaster recovery. AB - A disaster recovery plan (DRP) defines the scope of restoration, establishes responsibilities and lists specific actions to be taken after the disaster. Although not actually involved in a DRP or computer systems, nurse managers must understand the steps involved and identify and communicate nursing's requirements. PMID- 8718102 TI - Testing an electronic documentation system. AB - Testing of newly automated clinical documentation system functions in patient care areas is integral to successful automation. Trialing the new functions in the patient care environment can be accomplished by use of alpha and beta patient care areas. Throughout the trialing of functions, involvement and feedback from patient care area managers and staff is crucial for successful automation. PMID- 8718103 TI - Preparing for electronic documentation. AB - Anticipating JCAHO review within 1 year, the nursing department developed and implemented a staff-developed manual system using Focus charting. Forms were developed that would be easily transferred to computerization. PMID- 8718104 TI - 1996 software guide. PMID- 8718105 TI - What is the future of the OR director? AB - So, if there are still some nurses out there who want to get into administration, here's some good advice: "Pick your mentors, get a master's degree early in your career, get involved in your professional association and the sooner you commit, the better." There are still a lot of opportunities for OR managers, and the better ones will flourish in tomorrow's OR. The efficient and forward-looking OR managers will always do well and the well-managed ORs will survive! PMID- 8718106 TI - An ergonomics program for the health care setting. AB - The basic elements needed to establish a formal ergonomics program are addressed: work site analysis, hazard prevention, training and education and medical management. Occupational risk factors, both environmental and workplace, also are identified. PMID- 8718107 TI - Why good people do bad things. AB - The very context of organizational decision making tests autonomous moral agency for managers, and the moral price they pay may be great. Misunderstandings, miscommunication, fear and isolation all play a role in why good people do bad things. Self-knowledge, good communication, alternative support systems and management by walking around (MBWA) all help good people (who happen to be managers) avoid doing bad things. PMID- 8718108 TI - Perinatal bereavement. PMID- 8718109 TI - Journeying through labour and delivery: perceptions of women who have given birth. AB - OBJECTIVE: The aim of this study was to explore the essential structure of the lived experience of childbearing, as seen from the perspective of women who have given birth. DESIGN, SETTING, AND PARTICIPANTS: The phenomenological perspective of qualitative research theory guided the methodological approach to the study, in which interactive interviews were conducted with a purposeful sample of fourteen mothers of healthy babies in Akureyri and Reykjavik, the two most inhabited places in Iceland. FINDINGS: The metaphor of a woman journeying through labour and delivery was chosen to symbolize the lived experience of giving birth to a healthy baby. This encompasses four major categories: influences of circumstances and expectations before the journey's commencement; a sense of self during the journey which encompasses a sense of being in a private world, the sense of control, the need for caring and understanding and the need for a sense of security; the journey through labour and delivery itself; and finally the first sensitive hours of motherhood and the perception of the uniqueness of birth as a life experience at the journey's end. The study has the potential of increasing the knowledge and understanding of giving birth as a life experience, and therefore, has implications for midwives and nurses, as well as for women and their supporters. KEY CONCLUSIONS: The lived experience of giving birth is a powerful life experience which is coloured by circumstances and expectations of the woman, her sense of self during the journey, the journey itself, as well as the first sensitive hours of motherhood. IMPLICATIONS FOR PRACTICE: The study has the potential of increasing the knowledge and understanding of giving birth as a life experience, and therefore, has implications for midwives/nurse, as well as for women and their supporters. PMID- 8718110 TI - Antenatal 'booking' interviews at midwifery clinics in Sweden: a qualitative analysis of five video-recorded interviews. AB - OBJECTIVE: To describe antenatal 'booking' interviews as regards content and illuminate the meaning of the ways midwives and expectant parents relate to each other. DESIGN: Content analysis and phenomenological hermeneutic analysis of transcribed texts from five video-recorded antenatal booking interviews. SETTING: Midwifery clinics at five health centres in the context of Swedish primary care. PARTICIPANTS: Five midwives, five pregnant women (less than 14 weeks pregnant) and two expectant fathers. FINDINGS: A variety of content themes and ways of relating were found. Combined themes of biomedical and obstetric content occurred as frequently as the sum of social, emotional, antenatal care and life-style themes. The midwives' ways of relating formed two main themes; considering and disregarding the uniqueness of the expectant parents. The midwives directed the interview through their choice of content themes and the way they related to the expectant parents. The expectant parents mainly shadowed the midwives' content themes and ways of relating. The expectant fathers seemed like strange visitors in the women's world. Two perspectives of antenatal midwifery care, obstetric and parental, operated alternately and in competition within the interviews. KEY CONCLUSIONS: The content and the ways of relating within the interviews seem to be connected and could be understood in the light of Buber's writings on dialogue. IMPLICATIONS FOR PRACTICE: The findings provide a basis for reflection on the education of midwives and the planning, training and implementation of midwifery care at antenatal 'booking' interviews. PMID- 8718111 TI - An exploration of midwives' attitudes to research and perceived barriers to research utilisation. AB - OBJECTIVE: To explore midwives' views about research and their perceived barriers to research utilisation. SETTING: Thirty-two midwives from four midwifery units in the north west of England. The units ranged in size from a small district unit (52 midwives, and 1200 deliveries per annum), to a large regional centre (290 midwives, 6500 deliveries per annum). METHODS: Midwives' opinions concerning research were explored using focus group interviews; within the interviews midwives were asked to discuss how they viewed the relevance of research to midwifery care, the constraints which they felt prevented them from delivering research-based care and existing and potential methods of disseminating research. FINDINGS: There was a consensus among the midwives that they aspired to deliver research-based care. However, there are clearly a number of barriers preventing this. Research was poorly accessible to most midwives, both in terms of its physical location and complexity. Furthermore, midwives felt they lacked the knowledge and skills to appraise research, and lacked the confidence to judge when research should be implemented. CONCLUSIONS: The current trend to demedicalise childbirth demands that midwives become proficient users of research. However, the means of disseminating research findings to midwives in the north west of England does not meet their needs. PMID- 8718112 TI - A comparative study of breast feeding after traditional postnatal hospital care and early discharge. AB - OBJECTIVE: To provide quality assurance for the care plan and working structure within the early discharge unit at the Women's Clinic, Central Hospital, Helsingborg, Sweden. DESIGN: Survey, using postal questionnaire. SETTING: The Women's Clinic, Central Hospital, Helsingborg, Sweden. PARTICIPANTS: 304 women with babies of six months of age, delivered at the Central Hospital, Helsingborg, between September and December 1993 and who, together with their baby, met the criteria for early discharge. Early discharge is generally defined in Sweden as discharge before 72 hours postpartum. MEASUREMENTS AND FINDINGS: Of the participants 41% chose early discharge (ED) and 59% chose traditional hospital care (THC). Four groups were studied for breast feeding frequency - THC primiparae, ED primiparae, THC multiparae and ED multiparae. Further division was made for breast feeding at 2, 4 and 6 months of age. The four main groups were examined for demographic differences. A difference was found in education level; early discharge mothers had a lower level of education than THC mothers. No significant difference was found for the frequency or duration of breast feeding between the early discharge and the traditional hospital care groups, despite the higher education level in the traditional hospital care group. IMPLICATIONS FOR PRACTICE: A possible explanation for this finding is that a care plan aimed at supporting the individual's responsibility and participation, providing relevant knowledge and a subliminal communication of trust in the competence of parent and child, is of particular significance for women with less education. The presence and participation of the baby's father at an early stage may also be a factor. PMID- 8718113 TI - Women's evaluations of their experience with fetal intrapartum oxygen saturation monitoring and participation in a research project. AB - OBJECTIVE: To evaluation women's perceptions of their participation in a research project in labour and their experience with fetal oxygen saturation monitoring. DESIGN: Survey using a questionnaire of women's rating of ten aspects of their experience with the project. SETTING: Royal Women's Hospital, Brisbane, Australia. PARTICIPANTS: Women who had participated in clinical observational studies of fetal intrapartum oxygen saturation monitoring. FINDINGS: 78 questionnaires were distributed and 77 returned. Overall, women evaluated their participation in this research project favourably, in terms of both their involvement in the research study and the use of fetal oxygen saturation monitoring. Several women commented that they appreciated the additional support they received form the research midwife. KEY CONCLUSIONS: Responses indicated that the women surveyed appreciated the opportunity to participate in research, were not averse to this new technology, and benefited from the perceived additional human contact and support they received. The high response rate to the questionnaire indicates that childbearing women are willing to be included in assessing the appropriateness of new birth technologies that may affect the well being of women and babies in the future. PMID- 8718114 TI - [Educational processes for organizational change]. PMID- 8718115 TI - [Hospital equipment]. PMID- 8718116 TI - [Patients judge home visits]. PMID- 8718117 TI - [Acute trichinosis. Nursing care]. PMID- 8718118 TI - [The underside of the tongue]. PMID- 8718119 TI - [The care relationship in Alzheimer's disease]. PMID- 8718120 TI - [The nursing process as a practical working instrument]. PMID- 8718121 TI - [Being a nurse in a care team]. PMID- 8718122 TI - [Nutrition in the early medieval period. Basis of care]. PMID- 8718123 TI - [Enteral nutrition]. PMID- 8718124 TI - [Diabetes type II, non insulin-dependent. Group education]. PMID- 8718125 TI - Stroke. Professional issues. PMID- 8718126 TI - Performance-related pain. PMID- 8718127 TI - Staff at odds in scheme to help offenders. PMID- 8718128 TI - Separated by the system. PMID- 8718129 TI - Soft target. PMID- 8718130 TI - An education in bullying. PMID- 8718131 TI - Turning the league tables. PMID- 8718132 TI - The wax and wane of care levels. PMID- 8718133 TI - Distance no object for learning and treatment. AB - Telemedicine has up to now been mainly concerned with the application of new technology to the practice of medicine. However, there are potential applications for nursing and midwifery. This paper describes how it has been employed and looks at potential applications being investigated at the Institute for Telemedicine and Telecare at the Queen's University in Belfast. The use of video conferencing within a college of nursing is also explored. With the reorganisation of nursing education, institutions may need to coordinate activities over a wide geographical ares. To date, the use of video links from the nursing and midwifery school in Nottingham has resulted in less time spent travelling between sites for staff and students. PMID- 8718134 TI - Close encounters of a video kind. PMID- 8718135 TI - The developing role of the forensic community nurse. AB - Mental health professionals are increasingly concerned with the consequences of clinical judgement errors. Such concern has led to an increase in requests for specialist advice from forensic mental health care teams. Consequently, the demand for forensic community mental health nurses (FCMHNs) to provide specialist advice and reports has increased. Such advice can be significant in planning and delivering care. PMID- 8718136 TI - Using qualitative research in community nurse casework. AB - This paper explores how methods and skills frequently employed in qualitative research can be used by community nurses to promote an interpretation of the setting that is valid and client-focused. It discusses the construction and use of casenotes, the development of theoretical sensitivity, employing native language terms, and looks at the importance of a critical awareness of additional documentary evidence. PMID- 8718137 TI - Symbolic success. PMID- 8718139 TI - Second sight. PMID- 8718140 TI - Tutored to teach. PMID- 8718138 TI - United voices. Interview by Michael Walker and Chris Hart. PMID- 8718141 TI - A poor reference. PMID- 8718142 TI - Pressure sore assessments. PMID- 8718143 TI - Reducing the pressure. PMID- 8718144 TI - The use of hydrocolloids. PMID- 8718145 TI - Don't accept the hype. PMID- 8718146 TI - Reporting sexual harassment in an undergraduate nursing program. PMID- 8718147 TI - Community experience in mental health nursing. PMID- 8718148 TI - Introducing case management in the associate degree curriculum. PMID- 8718149 TI - A new image of nursing faculty participation in undergraduate education. PMID- 8718150 TI - Multicultural education: a second look. PMID- 8718151 TI - Integrating electronic conferencing to enhance problem solving in nursing. AB - The authors describe how a computer-mediated conference was integrated into a baccalaureate nursing program clinical course. They discuss methods used in implementing the conference, including a technical review of the software and hardware, and methods of implementing and monitoring the conference with students. Examples of discussion items, student and faculty responses to posted items, and responses to use of the computer-mediated conference are included. Results and recommendations from this experience will be useful to other schools integrating computer-mediated conference technology into the nursing school curriculum. PMID- 8718152 TI - Methodologic trends in the healthcare professions: computer adaptive and computer simulation testing. AB - Assessing knowledge and performance on computer is rapidly becoming a common phenomenon in testing and measurement. Computer adaptive testing presents an individualized test format in accordance with the examinee's ability level. The efficiency of the testing process enables a more precise estimate of performance, often with fewer items than traditional paper-and-pencil testing methodologies. Computer simulation testing involves performance-based, or authentic, assessment of the examinee's clinical decision-making abilities. The authors discuss the trends in assessing performance through computerized means and the application of these methodologies to community-based nursing practice. PMID- 8718153 TI - Health education via interactive television. AB - Providing health education in rural areas is a challenge because of the time and cost involved in travel to serve small groups of clients in their own towns. Baccalaureate nursing students provided health education to faculty and staff members in five rural high schools simultaneously through the use of interactive television. PMID- 8718154 TI - Home health visits in a nursing laboratory: preparing nursing students for their initial community experience. PMID- 8718155 TI - Dressing up nursing diagnoses: a critical-thinking strategy. AB - Teaching nursing diagnoses to beginning students who have no clinical experiences challenges educators to use their creative energies in developing strategies that facilitate students' critical-thinking skills. Dressing Up Nursing Diagnoses is a fun and creative classroom teaching strategy you can use during the Halloween season to help beginning students formulate nursing diagnoses. Two nurse educators describe their successful experiences with this unique teaching strategy. PMID- 8718156 TI - Clinical experiences in day care settings. AB - Day care settings offer many opportunities for students to participate in comprehensive primary healthcare and preventive health education. Additionally, day care sites provide experiences beyond those typically associated with well child healthcare. The author discusses practical issues for faculty members to address when seeking experiences for their students in day care environments. PMID- 8718157 TI - Test design: putting all the pieces together. AB - Preparing a nursing examination involves more than writing test items. The design of a test can contribute to measurement error and affect the validity and reliability of students' test scores. Nursing faculty should strive to minimize measurement error by following certain design rules when preparing a test. The author offers guidelines to help teachers design tests that will enable students to demonstrate their best performance. PMID- 8718158 TI - Respect: improving student writing. AB - Respect is central to the process of teaching writing, especially the respect that flows from asking students to write about their own personal experiences. The author offers suggestions on how to help students overcome their dislike of writing, address some of their writing deficits, and ultimately begin to enjoy writing. PMID- 8718159 TI - Debate: a teaching strategy to improve verbal communication and critical-thinking skills. AB - Debate is presented as a valuable learning activity for teaching critical thinking and improving communication skills. Included in the discussion are a brief history of the use of debate as a teaching strategy, the responsibilities of the teacher and learner when using debate in the classroom, and its many advantages for developing competencies in communication and critical thinking. PMID- 8718160 TI - Perspectives on dichotomous thinking. AB - Nursing education and practice often promote dichotomous thinking. There are serious limitations to this mode, as have been recognized by researchers in various fields. Nursing educators must overcome the entrenchment of dichotomous thinking in our teaching methods. PMID- 8718162 TI - Dementia. The role of the nurse. PMID- 8718161 TI - The poster presentation as an educational tool. AB - The authors describe their experiences in using the poster presentation to develop undergraduate nursing students in the scholarly role of professional nursing practice. The posters were used as a means for students to communicate their accomplishments in implementation and evaluation of an innovative clinical project. Methods for development, exhibition, and evaluation of the poster project are described. PMID- 8718163 TI - Pay up or pay the price. PMID- 8718164 TI - Yellow peril. PMID- 8718165 TI - Bumpy rides on the road to emigration. PMID- 8718166 TI - Caring to the end. PMID- 8718167 TI - Visions for the future. PMID- 8718168 TI - Radical overhaul. PMID- 8718169 TI - Flexible but focused. PMID- 8718170 TI - A fund of changes. PMID- 8718171 TI - From a client's perspective. PMID- 8718172 TI - Collaboration puts the ENB Higher Award into practice. AB - The ENB Framework for continuing professional education and Higher Award for nurses, midwives and health visitors was introduced in April 1992. The framework outlines 10 key characteristics to guide practitioners in the integration of theory and practice. This paper describes how a group of teachers and practitioners have applied the framework. They explain how it helps pull theoretical learning and practice together for practitioners. For educators, involvement with practitioners has helped them be more involved in practice. Three-way, or tripartite, tutorial work between the practitioner, the teacher and the facilitator in the clinical area is central to the success of this initiative. PMID- 8718173 TI - Aura photography: mundane physics or diagnostic tool? AB - Kirlian photography is often associated with the paranormal. Many people believe it records the auras of living objects and that it can be used as a diagnostic tool. This paper argues against these beliefs and maintains that there is a simple, scientific explanation of the Kirlian effect. PMID- 8718174 TI - Caring for the carers of people with mental health problems. AB - A ward manager has set up a forum where carers of relatives or friends with mental health problems can discuss and find solutions to their problems. In 1994, relatives and carers were invited to join the special hospital-based group, which met every fortnight. The ward manager was present in a facilitative role, but a 'bottom-up' approach focused on the needs and wishes of participants. In this paper, the author stresses the importance of respecting participants' views, and communicating complaints to the relevant hospital departments. PMID- 8718175 TI - Making health services more accessible to younger people. AB - High rates of teenage pregnancy and the demands of younger people for more accessible health care were the driving forces behind a series of initiatives to develop more appropriate services. The developments in a rural area of Tayside involved inter-agency working and young people's ongoing involvement. PMID- 8718176 TI - Whose job is it to care? PMID- 8718177 TI - Completing the whole. PMID- 8718178 TI - Weaning: dietary advice for parents. PMID- 8718179 TI - Us and them. PMID- 8718180 TI - Do race and ethnicity influence perceptions of health care practitioners? PMID- 8718181 TI - A clinical experience that convinced me to become a nurse. PMID- 8718182 TI - [Behavioral and cognitive therapies. Potent therapeutic tools]. PMID- 8718183 TI - [Phobic disorders]. PMID- 8718184 TI - [Obsessive-compulsive disorder]. PMID- 8718185 TI - [Social phobias]. PMID- 8718186 TI - [Troubles of the digestive tract. Role of the nurse in the hospital department]. PMID- 8718187 TI - [Behavioral therapies. Activities outside the hospital]. PMID- 8718188 TI - [Addiction: shame or sin?]. PMID- 8718189 TI - [Smoking prevention]. PMID- 8718190 TI - [The effects of a growing medicalization. Nursing records as an example]. PMID- 8718191 TI - [The sexuality of mental patients]. PMID- 8718192 TI - [You love me, you do not]. PMID- 8718193 TI - [Care of a severely burned child]. PMID- 8718194 TI - [Nicolas: "My home is the intensive care unit"--report on the care of a long-term patient]. PMID- 8718195 TI - [Protection and hygienic measures in respiratory syncytial virus infections]. PMID- 8718196 TI - [Whooping cough--complications and prevention of whooping cough in children]. PMID- 8718197 TI - [The foreign child in the hospital]. PMID- 8718198 TI - [Toxic and less toxic plants. 27. Datura]. PMID- 8718199 TI - [Current aspects of newborn infant nutrition]. PMID- 8718200 TI - [Law for the protection of mothers--the main item of labor legislation for women]. PMID- 8718201 TI - [Memories of Prof. Boehncke and the Hamburg Pediatric Hospital Hochallee--lecture at the interdisciplinary seminar "Fundamentals of Ethics in Medicine" on 4/25/1996 at the Hamburg-Eppendorf University Hospital]. PMID- 8718202 TI - [Eulogy for Professor Dr. Helmut Boehncke]. PMID- 8718203 TI - [Nursing and care of patients with cystic fibrosis]. PMID- 8718204 TI - Background & cosmology of Chinese diet therapy in childbearing. PMID- 8718205 TI - Infant feeding--midwives in the firing line. Part 2. PMID- 8718206 TI - The place of policy in a professional organization. PMID- 8718207 TI - Respect your time. Interview by Elizabeth Duff. PMID- 8718208 TI - Questions & answers in industrial relations. PMID- 8718209 TI - Postnatal depression: focusing on a neglected issue. PMID- 8718210 TI - A mentorship program for pre-professional nurse majors. PMID- 8718211 TI - Fathering as the pregnant father. PMID- 8718212 TI - [Epileptic crisis. The epilepsies]. PMID- 8718213 TI - [Epileptic crisis. The epileptic patient]. PMID- 8718214 TI - [Epileptic crisis. The treatments]. PMID- 8718215 TI - [Epileptic crisis. A place for care and education. The specialized institute Le Parc de l'Abbaye]. PMID- 8718216 TI - [Epileptic crisis. The place of the nurse]. PMID- 8718217 TI - [Epileptic crisis. Psychomotor performance]. PMID- 8718218 TI - [Epileptic crisis. The institutional psychiatrist. His role]. PMID- 8718219 TI - [Epileptic crisis. Psychotherapy for children. Institutional work and epilepsy]. PMID- 8718220 TI - [Epileptic crisis. Quality of life of young epileptics]. PMID- 8718221 TI - [Mourning in children]. PMID- 8718222 TI - [Understanding before learning. Theoretical basis of normal infant nutrition up to the age of 18 months]. PMID- 8718223 TI - [Homage to George Vacola. Quasimodo or the deficit in loving]. PMID- 8718224 TI - The future of nursing?. Interview by Dina Leifer. PMID- 8718225 TI - Lies, damn lies, and HNS funding figures. Interview by Pat Healy. PMID- 8718226 TI - Go for it! PMID- 8718227 TI - Use complaints. PMID- 8718228 TI - Getting the message across. PMID- 8718229 TI - Occupational stress: paying up. PMID- 8718230 TI - Sharing a vision. PMID- 8718231 TI - Health over the phone. PMID- 8718232 TI - Use of nasogastric tubes for elective laparotomy. PMID- 8718233 TI - Group work for bereaved children: a team approach. AB - This article provides a review of the current literature relating to child bereavement. The author also describes the setting up, running and evaluation of group work for bereaved children, and examines how these experiences can help to inform future best practice. PMID- 8718234 TI - Cataract surgery and recent advances: a review. AB - This article summarises the causes, symptoms and management of cataract with a brief description of the techniques of cataract removal and implantation of an ocular lens, including the recently introduced procedure of phacoemulsification. The authors also discuss the anatomy and function of the lens, helpful in understanding the surgical procedure. PMID- 8718236 TI - Health assessment of people over 75 (continuing education credit). PMID- 8718235 TI - Living with rheumatoid arthritis: a phenomenological exploration. AB - The objective of this research was to elicit the experience of living with rheumatoid arthritis (RA) from the patient's frame of reference and to understand the global impact that this chronic illness has on social, psychological and physical well-being. A phenomenological methodology incorporating Colaizzi's procedural steps (1978) was used. Five main themes emerged from the research which highlighted the enormous impact that RA has on a patient's life. Nurses need to adopt an holistic approach to care and ensure that they understand the illness from the patient's perspective. PMID- 8718237 TI - Questions & answers in industrial relations. PMID- 8718238 TI - The childfree midwife. The significance of personal experience of childbearing. PMID- 8718239 TI - Diet therapy in the postnatal period from a Chinese perspective. Part 2. PMID- 8718240 TI - Midwives and money matters: a simple guide to savings & investments. PMID- 8718241 TI - Symphysis pubis pain. PMID- 8718243 TI - [The behavior of asthmatic patients. Prevention of crises between the ages of 18 and 65]. PMID- 8718244 TI - [Promotion of mental health--new attitudes]. PMID- 8718245 TI - [Nazi experience of euthanasia: a memory and a lesson]. PMID- 8718246 TI - [From beginner to expert]. PMID- 8718247 TI - [A Pandora's box? A methicillin-resistant Staphylococcus aureus]. PMID- 8718248 TI - [Serving in Macao]. PMID- 8718249 TI - Child health nursing. PMID- 8718250 TI - The importance of negotiating a care plan. PMID- 8718251 TI - A resource centre for parents and professionals. PMID- 8718252 TI - Deliberate self harm in children and adolescents. PMID- 8718253 TI - A new retrieval service. PMID- 8718254 TI - Central line dressings: study of infection rates. PMID- 8718255 TI - Day case provision in a district general hospital. PMID- 8718256 TI - Are you listening? PMID- 8718257 TI - The pathophysiology of the pregnant father. PMID- 8718258 TI - [Internal partnership and quality of care]. PMID- 8718260 TI - [Voluntary workers and their associations. New actors in the health system]. PMID- 8718259 TI - [Partnership. The possible legal framework]. PMID- 8718261 TI - [Partnership and nursing schools]. PMID- 8718262 TI - [Staff education and university partnership]. PMID- 8718263 TI - [Psychologists at the Institute for Nursing Education]. PMID- 8718264 TI - [Cultural aspects of nursing]. PMID- 8718265 TI - [The Leonie Chaptal Foundation]. PMID- 8718266 TI - [Dimensions of teaching]. PMID- 8718267 TI - [Teaching plan for health administrators. At the Poissy Nursing School]. PMID- 8718269 TI - [Values and care. Total admission of a patient: from dream to reality]. PMID- 8718268 TI - [Nurses and writing]. PMID- 8718270 TI - [A teaching space: the multimedia]. PMID- 8718271 TI - [Letter to a supervisor in a home for the aged]. PMID- 8718272 TI - A very dangerous patriarchal movement to push mothers to breastfeed. PMID- 8718273 TI - Most sexually active teenagers would have unprotected sex. PMID- 8718274 TI - A management problem. PMID- 8718275 TI - Whose job is it anyway? PMID- 8718276 TI - Helping ourselves. PMID- 8718277 TI - Sharing skills. PMID- 8718278 TI - Ethics alert. PMID- 8718279 TI - With a view to change. PMID- 8718280 TI - Plan both halves of your career. PMID- 8718282 TI - Developments in hospital and community links. AB - This is the first of our reports from the King's Fund Development Centre, highlighting units which have developed experience in a particular aspect of nursing care and are keen to disseminate their work. This report reviews work being done in nursing units to improve the links between primary and secondary care, and is based on a Nursing Development Network day held last year. PMID- 8718281 TI - Debates and dilemmas. PMID- 8718283 TI - Development of specialist dermatology nurse prescribing. AB - In 1993, nurses on the dermatology unit at the Cumberland Infirmary, Carlisle, undertook a prescribing trial in order to assess and evaluate the accuracy and safety of nurse prescribing in this specialty. The aim was to demonstrate a positive outcome with evidence which would provide a sound, logical argument to explore the legal right of experienced dermatology nurses to prescribe where necessary within the unit. Five nurses and three senior house officers (SHOs) took part in the study which compared their theoretical prescriptions for treatment with two consultants who actually prescribed. Sixty one per cent of treatment prescribed by the SHOs was of the same type and potency as the consultants' choice. Nursing staff, however, accurately prescribed 80 per cent of the topical treatment the patients received. The authors conclude that nurses should be given more prescribing rights. PMID- 8718284 TI - Using 'a Vision for the Future' targets to change practice. AB - This article reviews the progress of a gynaecology unit towards meeting the targets set by A Vision for the Future (DoH/NHSME 1992). The authors describe how the targets have helped change practice and detail the initiatives developed to facilitate such change. PMID- 8718285 TI - Early discharge schemes for hysterectomy patients. AB - This review looks at the needs of hysterectomy patients discharged early from hospital. The author explores the concept of early discharge schemes and evaluates whether they can be safe, cost effective and provide quality care. Reflection on these factors suggests that an early discharge scheme could be a way to provide choice, and fulfill expectations of quality, holistic care and cost effectiveness, while allowing the nurse to work as an expert practitioner. PMID- 8718286 TI - Promoting urinary continence (continuing education credit). PMID- 8718287 TI - [Rheumatoid arthritis. Epidemiology and diagnosis]. PMID- 8718288 TI - [Rheumatoid arthritis. The disabilities]. PMID- 8718289 TI - [Rheumatoid arthritis. Medical and surgical treatment]. PMID- 8718290 TI - [Rheumatoid arthritis. Exercise therapy]. PMID- 8718291 TI - [Rheumatoid arthritis. Interdisciplinary care]. PMID- 8718292 TI - [The French Association of Polyarthritis Patients]. PMID- 8718293 TI - [The evolution of language in nursing 1975-1990]. PMID- 8718294 TI - [The field of ethics for nurses]. PMID- 8718295 TI - [Evidence of social links in the hospital or from the lost individual to the informed one]. PMID- 8718296 TI - [A project for success]. PMID- 8718297 TI - ["Behind every insurance reimbursement there is a sick person"]. PMID- 8718298 TI - [Nursing education programs in Germany. 4. In Freiburg they study while nursing]. PMID- 8718300 TI - [Nursing visits: the patient is being actively involved in his care]. PMID- 8718299 TI - [Self-care for nurses. 4. Tips for summer--sun--leisure time]. PMID- 8718301 TI - [Movement in the care of patients with osteoporosis. Idleness is the beginning of many troubles]. PMID- 8718302 TI - [Care of dying patients. 1. A seminar during training increases competence]. PMID- 8718303 TI - [Falsifying records is forgery of documents]. PMID- 8718305 TI - [Project development--quality management. The implementation is very time and personnel consuming]. PMID- 8718304 TI - [Problems in the financing of care: what is the social value of an old person]. PMID- 8718306 TI - [Nursing personnel provisions and organization of nursing services. Demonstrated in a surgery department]. PMID- 8718307 TI - Infant feeding--midwives in the firing line. Part One. PMID- 8718308 TI - Infant nutrition perspectives. PMID- 8718309 TI - Royal Society of Medicine forum on maternity & the newborn--antenatal care reconsidered. PMID- 8718310 TI - Questions & answers in industrial relations. PMID- 8718311 TI - Midwives and money matters: a simple guide to superannuation. PMID- 8718312 TI - No one is indispensable. PMID- 8718313 TI - Attitudes to drug users according to staff grade. AB - The attitudes of professionals working with intravenous drug users are likely to be affected by clinical grade. Perceptions of service delivery to intravenous drug users are likely to be affected by status. Senior professionals tend to be more positive in their attitudes towards intravenous drug users. Attitudes and perceptions of service delivery are very important in determining the standard of care delivery. PMID- 8718314 TI - Using complementary therapies: acupuncture. AB - Acupuncture is a traditional Chinese healing system designed to achieve and maintain harmony. Needles are applied to points in 12 meridians, each of which links to an organ. Nurses are often asked about complementary therapies and need to be familiar with the theory. PMID- 8718315 TI - Evaluating asthma patient inhaler technique. AB - People with asthma need to be shown correct inhaler technique at initial diagnosis. Doctors, nurses and pharmacists are in a good position to demonstrate inhaler technique and make regular checks. Information and counselling for people with asthma would be helpful. PMID- 8718316 TI - Risk reduction strategies in the control of AIDS. AB - Early HIV/AIDS public health strategies appeared to have limited effect. Government policies altered in response to complacency from the general public. Nurses with a good knowledge base are ideally placed to target vulnerable groups, such as older people, for risk reduction strategies. PMID- 8718317 TI - Changes in the roles and responsibilities of nurses. AB - There is still confusion over expanded and extended nursing roles. Role extension often involves a task-centred approach to adopting traditional medical responsibilities. The "augmented role' describes a way of legitimising this extension of the nursing role. PMID- 8718318 TI - Therapeutic consistency following brain lesions. AB - An understanding of neuroplasticity is essential in the provision of rehabilitation for neurological lesion. Nurses should provide and ensure consistency of care as part of the multidisciplinary team. PMID- 8718319 TI - Dressings available on the Drug Tariff. PMID- 8718320 TI - Emollient applications for chronic skin problems. AB - Emollients soothe and hydrate the skin. These products are the mainstay of treatment for contact dermatitis. Nurses do not need to be experts in dermatology but they do need to understand the principles and be able to advise on treatment options. PMID- 8718321 TI - Inter-rater reliability of risk-assessment scales. AB - A local attempt to produce a pressure sore risk-assessment scale with greater inter-rater reliability than the Waterlow, Braden or Douglas scales failed despite the new scale's popularity with staff. PMID- 8718322 TI - The benefits of an i.v. team in hospital practice. AB - I.v. teams can benefit the patient, the hospital and the nursing profession. I.v. teams reduce fear, non-compliance and infection. Standardised procedures lead to safe and cost-effective care. PMID- 8718323 TI - Using a computer-linked fax in patient discharge. AB - Computers can send discharge information instantly to the appropriate agencies. Discharge information can be generated from existing hospital records, saving time and resources. PMID- 8718324 TI - A supportive partnership for CAPD patients. AB - CAPD patient education is a continuous process. The renal training centre maintains close relationships with patients' community CAPD nurses. A high nurse/patient ratio allows time to deal with individual problems. PMID- 8718325 TI - Clinical supervision: UKCC must be more proactive. PMID- 8718326 TI - Susan's breast reconstruction: a case study and reflective analysis. AB - This case study describes and analyses selected aspects of the nursing care of Susan during her stay on a plastic surgery unit. The necessity for her breast reconstructive surgery following mastectomy some years earlier is discussed, along with the importance of selection of the appropriate operation and preoperative preparation. A retrospective analysis of aspects of the nursing care is then undertaken, giving insights for possible improvements and role developments. Susan left hospital with a new body shape and was beginning to adapt to her altered body image. Her continued recovery and the restoration of her 'ideal' body image are discussed. Finally, the value of case studies is considered along with an outline of their potential as a tool in professional practice and education. PMID- 8718327 TI - Impact of poor record-keeping on leg ulcer care in general practice. AB - The number of practice nurses has substantially increased since the introduction of the new GP contract and the implementation of the White Paper Working for Patients. Unlike district nurses and health visitors, practice nurses frequently do not keep nursing records. This project investigated the impact of poor record keeping on patient care and on the professional identity of practice nurses. Results show that few practice nurses keep their own records, but that those who do appear to offer a more holistic model of care. The project also identified links between poor standards of record-keeping and unsafe practice, suggesting a need for further research into the recording of nursing activity in general practice. Factors influencing the ability of practice nurses to work collaboratively as a professional group are also explored. It is suggested that health commissions have a role in supporting the professional development of practice nurses to ensure good standards of patient care. PMID- 8718328 TI - Critical care pathway development: the way forward. AB - In many NHS trusts throughout the UK, critical pathways are being used as a method of managing patient care, enabling trusts to ensure that patients receive appropriate, high quality, cost-effective care. At the same time they can meet some of the requirements of The Patient's Charter, such as involving patients in decision making and keeping them informed during periods of treatment. This article first describes critical pathways and then discusses briefly the reasons why pathways were developed at the Guy's and St Thomas' Trust, and the stages of development before their introduction. PMID- 8718329 TI - Trauma nursing: the team approach. AB - The team approach to trauma care has demonstrated benefits for patients in terms of both quality of care and outcome. This article examines the current management and organizational strategies of trauma care and the effect these have on patient outcome. The four main roles of the nurse within the trauma team are discussed. Research has shown that during the resuscitation of patients only 26% of nurses felt confident that the care they gave was satisfactory. However, 81% of these nurses may be responsible for the care of patients with multiple injuries at least once a week (Hamilton, 1991). Although medical staff have a vital role to play within the trauma team, this article will focus predominantly on the role of the nurse. PMID- 8718330 TI - Parent-held shared care records: bridging the communication gaps. AB - Specialist centres provide excellent treatment and expert care, but the best interests of children may not be served if they must always attend a hospital far from home. Systems of shared care have been developed to offer care nearer the family home whenever possible. Parent-held shared care records were introduced at a paediatric haematology/oncology centre in response to issues raised by parents and professionals concerning the management of shared care. These were set up to facilitate communication, teamwork and family involvement. Consultation took place within the multidisciplinary team to ensure that the design of the records met the needs of each group. During a multidisciplinary study day the records were introduced to professionals from shared care hospitals. Before formally incorporating these records into regular clinical practice they were piloted with a group of families. The records have become an accepted part of the service provided for family and professional partners in shared care. Formal evaluation of this innovation is currently taking place. PMID- 8718331 TI - Childhood cancer--a family crisis 1: the impact of diagnosis. AB - There have been substantial improvements in both treatment and survival of childhood cancer in recent decades. However, the diagnosis of cancer in a child still poses a threat to the whole family unit. In this article, the first of a two-part series, the nature, purpose and functioning of the family are explored to suggest how families may perceive a cancer diagnosis in a child. Review of the literature indicates that the level of threat correlates with indicators such as overall family reaction, information and communication, position within the disease continuum, prognosis and the presence of metastatic disease. PMID- 8718332 TI - Experimental designs. AB - This article discusses randomized clinical trials, double blind designs and factorial designs and describes and explains their main requirements. It is shown that the randomized clinical trial is not a different experimental design, but is one that reflects the purpose for and the scale on which the post-test only or pre-test/post-test designs are often used. A common method for the prevention of experimenter bias in clinical trials is the double blind trial. Factorial designs make the simultaneous manipulation of two or more independent variables possible, thus producing data about their effect on the development variable. PMID- 8718333 TI - Lyofoam. AB - Foam dressings provide an excellent healing environment for granulating wounds. Lyofoam is a polyurethane foam dressing that comes in a variety of presentations for differing wound types. Lyofoam is suitable for light to moderately exuding, granulating wounds; Lyofoam Extra, a new addition, for heavily exuding wounds; Lyofoam A, an all-in-one dressing, for granulating wounds; Lyofoam T, a standard Lyofoam dressing with a special cross-cut, for tracheostomy wounds; and Lyofoam C, which has a carbon layer, for malodorous wounds. Lyofoam is a well established, clinically proven wound dressing that promotes the ideal wound environment for healing. It has a low-adherent contact layer which leaves delicate granulation tissue undisturbed. It is also effective in the treatment of overgranulation. PMID- 8718334 TI - Individual vs collective activity: a primary nursing paradox. AB - Primary nursing has been proclaimed as a way of providing nursing care that benefits both the patient and the nurse. This article supports this view; however, a pragmatic approach is taken to identify some of the paradoxes and contradictions that may be encountered. These include: conflicts of loyalty; planning psychological care; empowerment of patients; empowerment of nurses; and competition. By using examples from practice the issues surrounding these concepts are explored, examining the impact both on practice and on nursing as an occupation. Nurses must address these paradoxes if they are to prevent conflict and frustration. The future goal of fully implementing primary nursing will, however, provide great opportunity and potential for improving patient care. PMID- 8718335 TI - All nurses are responsible for implementing research. PMID- 8718336 TI - Avoidable deaths. PMID- 8718337 TI - Approaches to the care of people with dementia. AB - Respect for an older person's autonomy may challenge assumptions about safety. Custodial care is rarely in the interests of the patient. Health problems in older people are often complex and stereotypes and labels are unhelpful Offering choices to people with dementia is worth while but not easy. PMID- 8718338 TI - Scalp cooling to prevent hair loss in chemotherapy. AB - Hair loss is a potentially distressing side-effect of several cytotoxic drugs. Scalp cooling can help prevent hair loss, but is time-consuming and uncomfortable. Patients should be able to make informed choices about such treatment. PMID- 8718339 TI - Foot-care education in patients with diabetes. AB - Patients with NIDDM are at risk of serious foot problems. Patient education needs to be continued long after diagnosis and initial education. The risk of complacency must be recognised, especially when there is a high rate of co existing conditions. PMID- 8718340 TI - Neurodegenerative diseases: sustaining hope. AB - The concept of hope is associated with coping and adaptation. Maintenance of hope is a fundamental part of neuroscience nursing. Hope and coping must be important areas of concern to nurses working with patients who have neurodegenerative disorders. PMID- 8718341 TI - Supporting children with cystic fibrosis in school. AB - Children with cystic fibrosis need support to benefit from a normal education. Liaison between the specialist nurse and school staff--teachers and nurses--can enable the school to provide this support. PMID- 8718342 TI - Factors inhibiting the use of research in practice. AB - Several factors help to perpetuate the research/practice gap. Career opportunities for nurses in research may lead to better use of findings. PMID- 8718344 TI - Conflicts in care. PMID- 8718343 TI - Cranberry juice. PMID- 8718345 TI - Altered body image in patients on CAPD. AB - CAPD can restore independence but can compromise body image. Meeting others with similar experiences can help some patients. Nurses are responsible for preparing patients adequately before they embark on CAPD. PMID- 8718346 TI - Temperature measurement devices. AB - Temperature measurement contributes towards an objective indication of patient status. In-service education can enhance the accuracy of temperature measurement. The measurement of body temperature is traditionally viewed as a nursing responsibility. PMID- 8718347 TI - The lecturer-practitioner role in mental health. AB - The lecturer-practitioner role can be a means of supporting students and making education relevant to practice. The professional credibility--and skills--of the post-holder can be enhanced by this link with practice. It is vital to ensure that the clinical aspects of the post are not neglected. PMID- 8718348 TI - Supporting family carers: the key to successful long-term care? PMID- 8718349 TI - Factors influencing nurses' analgesia decisions. AB - The first article in this two-part series (Vol 5(13): 778-84) presented an empirical study of nurses' and patients' pain following surgery. Results indicate that nurses give consistently lower pain ratings for patient's pain than do patients. The main implication of this research is that nurses continue to underestimate the severity of pain suffered by patients postoperatively. This article considers a second empirical study, the objectives of which were to determine which methods nurses use to assess and monitor pain relief and which factors influence their decision to give or withhold analgesia. Results reveal that while nurses claim to rely on patients' self-reports of pain, the most influential factors in the decision whether or not to give analgesia are the dosage, type and frequency of drug prescribed. The main reason for withholding analgesia is that the patient reacted adversely to the medication. Student nurses placed more emphasis than trained nurses on the timing and amount of analgesia given. Nurses had a good knowledge of pain assessment tools, although few had used them. PMID- 8718350 TI - Caring for the adolescent diabetic. AB - The management of insulin-dependent diabetes mellitus is a challenge for the adolescent patient, his/her family and the health-care team supporting them. If treatment is to be successful and the longer-term complications of diabetes averted or delayed, nurses need to understand the various psychological and physiological factors that contribute to the difficulties in maintaining glycaemic control during this life stage. This article examines some of these factors with a view to encouraging nurses to adopt strategies that will improve patient confidence and coping skills in diabetic care. PMID- 8718351 TI - Psychology 2: message received and understood? AB - Human beings judge and make sense of their surroundings in two ways. Sensory organs supply information to the brain where it is processed and interpreted according to the idiosyncrasies of that individual's personality and experiences. Perception is the active process which integrates and coordinates these activities. Hence, any sensory message can be received and/or understood quite differently depending on the individual concerned. This article outlines how a knowledge of psychology can assist in the understanding of human perception, and how this can be applied to a wide range of nursing care settings and applications. The psychology of perception is of particular relevance to areas such as patient/client assessment, skilled communication, teaching and the provision of health education. PMID- 8718352 TI - Nurses' knowledge of pressure ulcer management in elderly people. AB - The nursing management of pressure ulcers has not been extensively reported in the Hong Kong context. This article reports the results of a survey of 34 Hong Kong registered nurses to determine their knowledge and practices in preventing and treating pressure ulcers in the elderly. The findings indicate that, overall, the nurses had a sound knowledge of the prevention and treatment of pressure ulcers, but that there were inconsistencies in their practices. This situation could be improved if standardized systems for assessing patients at risk of developing pressure ulcers and for grading (staging) these pressure ulcers were used. PMID- 8718353 TI - Validation therapy: a review of its contribution to dementia care. AB - There has been a great deal of interest in validation therapy (VT), an interactive technique and group therapy for people who are disoriented. This article reviews the strengths and development of VT and acknowledges Naomi Feil's contribution to the care of those with dementia. Techniques for communicating with people who are dysphasic are described, as are therapeutic interventions which address the devastating effects of memory loss on the sufferers' sense of self and identity. PMID- 8718354 TI - The biological basis of cancer. AB - People with cancer require sensitive nursing. They need information, explanations, encouragement, support and guidance through investigation, diagnosis and treatment options. One person in three may have a diagnosis of cancer sometime in his/her life, thus the nurse is faced with an increased probability of caring for a cancer patient during his/her career. In general, both the public and nurses have a negative view of cancer because of lack of knowledge about cancer. Knowledge of the fundamental pathophysiological aspects of cancer can increase understanding about the causes of cancer, its development and the rationale for treatment approaches. Ultimately, knowledge assists nurses in supporting and empowering their patients. PMID- 8718355 TI - Cica-Care: adhesive gel sheet. AB - Keloid and hypertrophic scars can be a complication of burns, surgery and other traumas. Cica-Care (Smith & Nephew) has been developed for the prevention and treatment of such scars. This product focus presents the clinical features of both types of scars, highlighting the difference between them, and outlines the treatment options which include surgery, pressure treatment and pharmacological treatments. The mechanism of action of Cica-Care is still uncertain, but it appears to soften, flatten and blanch scars. The specific care of the gel sheet, including skin acclimatization, is also described. PMID- 8718357 TI - Nursing's image: it is how you use your stethoscope that counts! PMID- 8718356 TI - The value of Peplau's theory for mental health nursing. AB - Recent changes in mental health care have necessitated reevaluation of Peplau's nursing theory. There is an expanding body of nursing knowledge and a debate is taking place regarding the difference between models and theories. Peplau's theory considers nursing as an interpersonal process between nurse and patient. The theory lacks empirical investigation and is not yet tested regarding the multidisciplinary team approach, which is largely based on the medical model. For the theory to gain credibility within the dynamics of the multidisciplinary team, it must be tested within a framework of robust research design involving various diagnostic groups and practice settings. Until such time, the theory's utility must seriously be questioned within contemporary mental health practice. PMID- 8718358 TI - Critical incidents and PREP. PMID- 8718359 TI - Using a database for managing interview data. AB - The definition of a database includes any structured collection of information. A computer database can be used to store transcripts, find particular entries and to extract subsets of data. PMID- 8718360 TI - The distribution of resources in health care. AB - Perceived advances in medicine lead to raised public expectations of care. Resources are finite, but governments can choose how they spend health service funds. Decisions on health "priorities' have implications for nursing practice as well as patient choice. PMID- 8718362 TI - Decision-making in the care of cavity wounds. AB - Good cavity-wound management requires a team approach. Nurses seeking autonomy may not recognise the responsibility that goes with it. There is still confusion and ignorance about wound dressings and agents used for cavity wounds. PMID- 8718361 TI - Children with cancer: the needs of siblings. AB - Siblings have been identified as the most neglected of all family members during serious childhood illnesses. Siblings of children with cancer can experience great stress. Nurses can support the whole family and encourage positive coping mechanisms among siblings. PMID- 8718363 TI - Working in a team. PMID- 8718364 TI - A nutritional assessment tool for older patients. AB - Older people are at increased risk of poor nutritional status. Nutritional support can improve the outcome of medical and surgical treatments and reduce length of hospital stay. Assessment is complex owing to the multifactorial nature of nutritional risk. An easily administered nutritional screening tool can be incorporated into the nursing assessment of older patients. PMID- 8718365 TI - Handwashing solutions. AB - Handwashing is the most vital of all nursing procedures. Nurses can influence practice and purchasing decisions in handwashing. All the costs of handwashing need to be considered when choices are made. PMID- 8718366 TI - The first assistant role in PEG tube insertion. AB - Nurses are under pressure to take on junior doctors' tasks. Expansion of the nurse's role should be of benefit to patients and professionally appropriate. The development of the first assistant role in PEG may be an avenue for expanding nursing. PMID- 8718367 TI - An audit of mattresses in one teaching hospital. AB - The poor state of some hospital mattresses has been associated with the formation of pressure sores and the spread of infection. Nurses need to be aware of potential problems with old, faulty or poor-quality mattresses. Mattresses must be checked regularly. PMID- 8718368 TI - Room service. PMID- 8718369 TI - The physiological effects of alcohol misuse. AB - Alcoholism is costly both in human and monetary terms. A thorough nursing assessment will lead to early detection and treatment. Further tissue damage and suffering can be reduced by early intervention. PMID- 8718370 TI - The nurse as significant other for surgical patients. AB - Significant others are a major influence on a patient's self-worth. For hospital patients, a nurse may become a significant other. The nurse's initial reactions, especially after major surgery, have the potential to influence recovery. PMID- 8718371 TI - Can a nurse be a patient's advocate? PMID- 8718372 TI - The concept of hope in life-threatening illness. AB - Achievable goals are important in fostering hope. The presence of hope is an important component of spiritual well-being. Being unrealistic may help the patient to cope. Nurses need to help patients achieve a balance between hopelessness and unrealistic hope. PMID- 8718373 TI - The role of the specialist nurse in breast cancer. AB - Psychosocial support is vital for women with breast cancer. The breast care nurse performs a crucial role in this support. PMID- 8718374 TI - Sociology and nursing. AB - Changes in society and in patterns of disease have led to changes in nursing practice and nurse education. Sociology can expand nurses' understanding of patients' needs in the context of the social environment. PMID- 8718375 TI - Breaking bad news to patients with cancer. AB - For the patient with cancer, bad news can occur several times between diagnosis and death. Patients expect honesty from professionals. Patients may choose to face up to bad news or actively deny what is happening. Nurses need to be able to deal with the patient's emotional response to bad news. PMID- 8718376 TI - A pressure sore risk scale for use with older people. AB - Identifying patients at risk can reduce the costs of preventing and treating pressure sores. Many accepted risk-assessment scales tend to over-predict risk. PMID- 8718377 TI - Managing a team. PMID- 8718378 TI - Counselling older people in residential care homes. AB - The emotional needs of older people living in residential homes are often neglected. A change of attitude is needed to address the counselling needs of this client group. Problems experienced by older residents usually include feelings of loss and bereavement. PMID- 8718379 TI - Infusion devices for ambulatory use. AB - Automated drug delivery is now indispensable in most hospital settings. The use of ambulatory infusion therapy in patients' homes is increasing. Nurses are often responsible for acquiring infusion devices. PMID- 8718380 TI - Compression hosiery. PMID- 8718382 TI - Physiologic effects of normovolemic anemia: implications for clinical monitoring. AB - Normovolemic anemia is commonly found in acute and critically ill adults. Compensatory physiologic effects of anemia are governed by the extent and rate of development of the anemia as well as by chronic or acute diseases that limit physiologic reserves. Increased metabolic demands that frequently accompany critical illness confound how patients will tolerate anemia. Clinicians should be aware of the physiologic responses to anemia, to monitor for intolerance and prevent negative outcomes. In this article, the author reviews specific physiologic responses occurring in adults with normovolemic anemia and describes appropriate clinical monitoring parameters. PMID- 8718381 TI - Blood conservation in acute care and critical care. AB - Blood conservation has evolved into an important issue in hospital-based medicine. Increased awareness of and worry about transfusion-associated diseases have prompted a focus on this important area. New technologies, including continuous intraarterial monitoring devices, microchemical technologies, new drug development (recombinant human erythropoietin and aprotinin) and intraoperative salvage techniques have made the concept of clinically important blood conservation possible. In this article, the authors review clinically important areas regarding blood conservation, which are subsequently detailed in this issue of AACN Clinical Issues. Emphasis is placed on the need for blood conservation in acute and critical care practice and the technologies available to achieve this goal. PMID- 8718383 TI - Current red blood cell transfusion practices. AB - The appropriate use of blood transfusions remains variable among health-care institutions and patient populations. Transfusion practices are discussed in this article in relation to medical practice guidelines and utilization review. Specific transfusion practices in the settings of intensive care, orthopedic surgery, and open heart surgery are reviewed. A new, promising approach to improving transfusion outcomes is the use of transfusion algorithms. Transfusion algorithms may prove especially useful if they incorporate point-of-care testing that is both physiologic and patient-specific for transfusion decisions. Transfusion algorithms are discussed and data presented for cardiac surgical adults. PMID- 8718384 TI - Preoperative autologous blood donation. PMID- 8718385 TI - Blood conservation in neonatal and pediatric populations. AB - Blood conservation in infants and children has benefits even beyond those seen with the adult populations. For instance, acquired blood borne diseases such as cytomegalovirus not only cause illness but also can have deleterious effects on the growth and development of infants and children. Decreasing blood transfusions is especially important in preventing sensitization over a lifetime, which may require further transfusion and even organ transplantation. A less striking benefit, but one equally as significant, is decreasing the occurrence of graft versus-host disease when blood conservation negates the need for multiple transfusions. The limitation of alternative transfusion practices in children and infants increases the benefits of blood conservation. Autologous blood donation may be an alternative to allogeneic transfusion in older children, but is not possible with neonates who may be born anemic and who experience a normal physiologic anemia during the first 2 months of life. Critical care nurses are instrumental in helping blood conservation by understanding blood salvaging techniques, including correct collection techniques, noninvasive monitoring, evaluation of diagnostic sample needs, and administration of erythrocyte stimulating factors. PMID- 8718386 TI - Intraoperative and postoperative blood salvage. AB - Surgical patients present unique opportunities for reducing allogeneic transfusions via perioperative blood salvage. Intraoperative cell saver techniques enable the collection of autologous blood from the surgical field and cardiopulmonary bypass circuit for return to the patient. Washing and hemoconcentration creates a product with an average hematocrit of 50% that carries normal erythrocyte survival, with minimal coagulation factors of platelets. Postoperative autotransfusion of shed blood from chest tubes or surgical drains enables additional blood conservation, reducing allogeneic transfusions by as much as 50%. Both intermittent and continuous autotransfusion systems using standard chest drainage equipment are available that require minimal setup and maintenance at the bedside. The hematocrit of shed blood is 20 25%, with depletion of clotting factors and platelet function. Potential advantages of perioperative blood salvage include the ready availability of large volumes of patient-compatible blood, with optimal oxygen-carrying capacity, that is virtually free from the risk of viral contamination. Specific indications, contraindications, and techniques for the salvage and administration of these vital products are presented. PMID- 8718387 TI - Diagnostic blood analysis using point-of-care technology. AB - Rapid analysis of selected laboratory tests is essential in the management of critically ill patients. These tests facilitate accurate diagnosis of clinical problems and the initiation and evaluation of appropriate therapeutic interventions. Instruments are now available to perform analysis at a site near the patient, or "point-of-care" testing. These instruments incorporate electrochemical and optical sensors capable of providing information on multiple analytes from a small sample of whole blood. In this article, the author explores the technologies behind point-of-care testing, as well as the advantages and disadvantages of in vitro, in vivo, and ex vivo modalities. PMID- 8718388 TI - The role of pharmacologic agents in blood conservation. AB - Pharmacologic agents are welcome therapeutic weapons in the fight to conserve blood. The goals of drug therapy cross a broad spectrum of potential areas. Blood conserving pharmacologic agents are aimed at 1) increasing blood production, 2) decreasing blood loss during active bleeding, and 3) preventing the breakdown of the formed fibrin clots. In this article, the author reviews the mechanisms of action, dosing, and adverse effects of these agents. The role of pharmacologic agents in blood conservation is significant because these agents have been shown to decrease transfusion requirements and provide significant cost savings. PMID- 8718389 TI - Nursing strategies to minimize blood loss associated with phlebotomy. AB - Blood loss associated with phlebotomy is significant in critically ill adults. Iatrogenic anemia may result and impose unnecessary stress on the cardiovascular and respiratory systems and may require allogeneic blood transfusions. Many strategies exist under nursing's direct control to decrease blood loss associated with phlebotomy. In the past, nursing effectively implemented many of these strategies in patients at high risk of anemia, such as pediatric, neonatal, transplant, or chronic renal failure patients, as well as patients who are Jehovah's Witnesses. Implementation of these strategies are needed for all critically ill patients because allogeneic blood transfusions carry infectious risk and because complications and chronic critical illness cannot be predicted reliably. Incorporation of these strategies into daily practice as well as the development of blood conservation programs represent imminent challenges for nursing. PMID- 8718390 TI - Current questions with regard to acute care nurse practitioner preparation and role implementation. AB - The role of the acute care nurse practitioner is being implemented in acute and tertiary care settings. As the role evolves, questions arise. Some of these questions relate to: 1) whether a need for the role has been clearly established; 2) whether the scope of acute care nurse practitioner practice has been identified distinctly; 3) the adequacy of educational preparation; 4) certification versus licensure; and 5) the nature of the relation between this role and other forms of advanced practice nursing. In this article, the authors identify and discuss some of these questions, provide information regarding current status, and postulate further potential resolutions. PMID- 8718392 TI - The acute care nurse practitioner in collaborative practice. AB - Successful collaborative practice between an acute care nurse practitioner and a physician impacts positively on patient care. Collaboration among disciplines can provide the patient with superior care that is organized, efficient, and prompt. Collaboration also gives each partner in the relationship a great deal of professional satisfaction. The acute care setting in Head and Neck Surgical Oncology provides an excellent environment for collaboration. The patient with head and neck cancer has many complicated medical and nursing problems that challenge the efficiency of this integrative practice model. In this article, the authors demonstrate the effectiveness of this practice model in the management of this patient population. PMID- 8718391 TI - The changing role of advanced practice nursing in a managed care environment. AB - Advanced practice nursing in a managed care environment--is this an oxymoron? Can these two phrases exist together? Do advanced practice nurses have a role in a managed care environment? As health-care systems continue to merge and managed care impacts the bottom line, advanced practice nurses are challenged to show their worth. In this article, the author reviews the impact managed care has had on the roles of two master's-prepared traditional clinical nurse specialists. Included are examples of how the roles were adapted to fit in the managed care setting. The current role of the advanced practitioners is reviewed, using the common subroles as a basis for discussion. The impact of managed care on each component of the clinical nurse specialist role is identified. The author also demonstrates that advanced practice nurses can have a valuable role in the managed care environment. PMID- 8718393 TI - Can we afford not to have clinical nurse specialists? AB - The clinical nurse specialist (CNS) is in an excellent position to lead and design the necessary changes required in acute, chronic, and home care settings to meet the economic and legislative agendas to reshape health-care delivery. In this article, the authors, through the use of examples, outline key quality characteristics of a successful unit-based, system-focused model of CNS practice. Clearly, the benefits obtained from the outcomes of a traditional CNS role make a significant contribution to the profession of nursing and to the public. Without the CNS role, the opportunity for system change and the development and nurturing of the front-line health-care worker may be lost. PMID- 8718394 TI - Ethical theories and principles. AB - Ethically sensitive situations and issues are not new in today's health care practice. They are increasing in frequency and complexity. For practitioners to face these challenges, they must not only understand their own values and beliefs, but understand the tools available to resolve these ethical dilemmas when they arise. Ethical theories and principles and decision-making models can assist the practitioner in obtaining resolution. PMID- 8718395 TI - Evolution of the American Nurses' Association Code for Nurses with interpretive statements: explications for perioperative nurses. AB - Although the need for ethical standards in the nursing profession has been recognized since as early as 1903, not until 1950 was a code of ethical principles adopted by American Nursing Association (ANA) members. The AORN Special Committee on Ethics was formed in 1991 to identify ethical issues in perioperative nursing and recommend educational strategies for addressing them. Formulation, publication, and distribution of the ANA Code for Nurses with Interpretive Statements--Explications for Perioperative Nurses resulted from the work of this committee. PMID- 8718396 TI - Ethics committees: their share in the advocacy role. AB - In the wake of escalating health care costs and advances in technology, health care faces an ever-increasing number of ethical dilemmas. Never before has the need for advocacy been greater, and never before has the role of ethics committees as advocates for patients and families been as imperative. PMID- 8718397 TI - Ethics and the nurse's commitment to political involvement: an educator's perspective on ethics and nursing. AB - Perioperative nurses concomitantly practice in the legal, moral, ethical, and political domains. Although the domains are often thought of as separate entities they are interwoven in practice. This report uses values as the basis of their interconnectiveness. Ethics and politics are presented as problem solving processes. Specifically, ethics is used to justify moral behavior/conduct and politics to resolve conflicts with perioperative and other health care settings. PMID- 8718398 TI - Caring and competence: perioperative nurses tell their stories. AB - One of the many challenges that face perioperative nurses is articulating the moral dimensions of our practice. The stories of practicing clinical nurses are used to articulate caring and competence, two essential components of the perioperative nursing practice. Analysis of patient's stories is essential to validate and safeguard the moral integrity and economic future of the registered professional nurse in the operating room. PMID- 8718399 TI - Ethics and human rights issues in perioperative nurses: a subsample of Maryland nurses. AB - The purpose of this study was to identify the ethics and human rights issues encountered by Maryland perioperative nurses in their practice. The most frequently occurring issues, the most disturbing issues, and content for ethics education are identified. Implications for education, management, and future research are presented. PMID- 8718400 TI - Ethical issues: a survey of perioperative nurses. AB - A convenience sample of 499 perioperative nurses was surveyed to determine what they perceive to be the most pressing ethical issues. Staffing patterns, informed consent, allocation of resources, and occupational risk were ranked as most important. Also surveyed were subjects' knowledge and application of the American Nurses Association (ANA) Code for Nurses with Interpretive Statements- Explications for Perioperative Nursing (ANA Code in AORN: Standards and Recommended Practices. Denver, CO, 1994. PMID- 8718401 TI - End-of-life decisions: the role of the nurse. AB - Nurses confront the clinical realities and controversies of end-of-life decisions in all practice settings and professional roles. It is imperative that nurses consider these issues and understand the ethical and professional parameters of practice. A code of ethics and professional position statements can provide guidance and support as nurses attempt to fulfill their professional obligations and deliver competent and compassionate end-of-life care. PMID- 8718402 TI - Cadaver organ donation and moral distress: a staff nurse's perspective. AB - Current advancements in medical science, such as the progress seen in the area of organ transplantation, brings with it many ethical dilemmas for which there are no precedents. Obtaining informed consent for cadaver organ donation requires perioperative nurses to confront the moral responsibility that they have to their patients, their patients' families, and to the nursing profession as a whole. The perioperative nurse must question his or her own moral and cultural beliefs, face their own fears of death, and confront societal misconceptions about brain death. This evolution is emotionally demanding and often stresses one's support systems. However, with self-discovery and education, it can also be very rewarding. PMID- 8718403 TI - Ethical issues in publication. AB - Nurses usually publish to disseminate pertinent information and research findings. Their articles and textbooks promote quality patient care and sound professional practice. Nurses may also publish to meet requirements for academic promotion and professional advancement. Ethical issues play a part in any phase of writing for publication. Veracity, justice, beneficence, and nonmaleficence are four of the ethical principles that pertain to aspects of authorship, peer review, and research in publication. PMID- 8718404 TI - Managing rumor and gossip in operating room settings. AB - The unique features of the operating room (OR) make it an ideal setting for the proliferation of gossip and rumor. Although not always negative, these "grapevine" communications can reduce productivity and work satisfaction. Hence, OR managers need to understand these forms of communication and prevent or control their negative consequences. The authors offer suggestions for undertaking this challenge. PMID- 8718405 TI - Conflict. AB - Conflict management is a major component of a nurse manager's role. How conflict is defined and subsequently approached can determine if its outcome is a positive, growth-enhancing experience, or, if instead, it will have lingering negative effects destined to resurface, provoking further conflict. When conflict progresses without effective intervention, others are drawn, or triangled in, and it becomes difficult to determine how, why, and with whom the conflict began. Approaches range from total avoidance to a fully invested, collaborative process of resolution. The collaborative response demands significant management involvement; however, its outcome can be the discovery of new and better practice opportunities, benefiting all involved. PMID- 8718406 TI - Manipulation: a manager's perspective. AB - Stressors of change affect staff, managers, and patient services. Destructive behavior, such as manipulation, further disrupts teamwork and flow of information within an organization, diverting energy that could be used positively for planning and problem resolution. Understanding manipulative behavior and corresponding interventions may assist the nurse manager in personal growth, anticipatory staff development, and proactive care in a changing environment. Manipulation does not promote efficient, efficacious care. PMID- 8718407 TI - Negotiation for the perioperative nurse: it's all up to you. AB - This article explores negotiation and related theories involved in the negotiation process. Philosophical views, stages, and various negotiation styles are discussed. Particularly, principled negotiation is addressed as a useful framework for conflict resolution. In addition, barriers, strategies, and skills inherent to the negotiation process are explored. Finally, a case application is presented to highlight the potential effectiveness of using successful negotiation tactics in the perioperative nursing setting. PMID- 8718408 TI - Elements of the employee selection process: interviewing operating room staff. AB - Selecting the right staff for employment in an operating room (OR) is critical to the success of any OR manager. Legal considerations limit the ability to gather certain information. However, by applying the correct skills and techniques, the manager can develop an information base that will lead to better decisions when selecting employees who can contribute to an efficiently run department. PMID- 8718409 TI - Employee misconduct: discipline or prevention. AB - Discipline is something that can only be imposed after an act of misconduct has already occurred, but managers can use strategies to prevent misconduct before it occurs. When an act of employee misconduct does occur, supervisors should know how to deal with the situation effectively. Management training and knowledge of appropriate administration of discipline will greatly enhance the probability that undesirable behaviors are changed and that when termination of employment becomes necessary, arbitration and litigation will not suspend, reduce, or reverse management's decisions. PMID- 8718410 TI - Job satisfaction: a possibility? AB - Registered nurse job satisfaction is one of the most written about topics in the nursing literature. Is it possible to achieve in an organizational setting? Can the administrative structures control this phenomenon? This article suggests that job satisfaction is strongly influenced by individual perceptions of certain key variables. In addition, job satisfaction is presented as a responsibility of each individual nurse. PMID- 8718411 TI - Practically perfect planning. AB - Planning is a routine human activity performed daily that often challenges confident professionals. The nursing process, adapted as a planning tool, provides the means to achieve institutional goals. The application of the nursing process to planning in one operating room setting resulted in the preparation of more than 1,500 surgical case supply preference lists in 5 weeks' time. The achievement facilitated timely implementation of an automated charging system for surgical cases through practically perfect planning. PMID- 8718412 TI - Financial perspectives for the perioperative nurse. AB - This article describes the preparation of the operating room budget and identifies six components that contribute to cost per case. The successful budget can be achieved by following a six-step budgetary process. The budget process is shown by sample tables. PMID- 8718413 TI - How nurses can make informatics work for them. AB - Nursing is one of the oldest clinical professions; however, it is not enough for today's nurses to be skilled in caring for the sick. They must also become efficient managers of their institution's resources. Emerging and ever-changing computer technology offers many tools that can help nurse managers plan, control, and evaluate the areas they control. This article discusses some the tools that nurses can use to make their jobs both easier and more effective. PMID- 8718414 TI - Using organizational culture as an agent of change. AB - Many health care organizations are considering or undergoing work redesign or other workplace change. The result is profound changes in the organizational culture. In addition, change is also required in the work unit cultures. Understanding organizational culture results in better planning and implementation of change. PMID- 8718415 TI - Introduction of solid foods to African American and Anglo American low-birth weight and full-term infants. AB - A secondary data analysis of 7,174 infants explores the use of cereal, fruits and vegetables, and meats with African-American and Anglo-American very-low-birth weight (VLBW), low-birth-weight (LBW) and term infants over the first five months after discharge. The first solid foods offered were cereal for African-American infants and fruits and vegetables for Anglo infants. There are statistically significant differences in the number of feedings offered by the two ethnic groups. During the last two-to-three months, African-American term and LBW infants received all solid foods more frequently than Anglo infants. Neither ethnic group followed current feeding practice recommendations on when to introduce solid food. PMID- 8718416 TI - Critical social theory and the domination of African American Women. AB - This historical reconstruction of the experiences of African American women in America from slavery to the present exposes the prevailing and enduring system of White male domination. From White men having control of their reproductive choices, to conspiracy to withhold the right to vote, African American women were victims of both sexism and racism. Later, as a result of the myth conceived by White sociologists of the super African American woman, further divisiveness became apparent in the African American home. As African American women took advantage of educational opportunities only to find that there was a dearth of similarly educated African American males to marry, increasing numbers of African American men were reported as parties to violent acts, drugs or illness. All of these variables are conjectured as impacting on the African American woman's experience. Lastly, data were presented depicting the increasing trend of African American women marrying White men, and the emergence of a more diverse workforce. It was concluded that economics serve as a catalyst for this change in human relations. PMID- 8718417 TI - Power perceptions of Black nursing faculty. AB - The purpose of this study was to determine the perceptions of Black nursing faculty members to the concept of power. This study sought to answer the question: How do Black nursing faculty members perceive power and how are they oriented toward power? The Cavanaugh Power Orientation Scale (1978) was used to survey a random sample of 100 Black nursing faculty members obtained from a national listing. The data were analyzed using descriptive methods of measurement, factor analysis, Pearson Correlation Coefficients and analysis of variance. An analysis of the data show that factors did not load of the same variables as had been shown in Cavanaugh's study. The presentation will discuss the descriptive data and the scores obtained on each of the six factors. It is important to understand the perceptions that Black nursing faculty members have about the concept of power because these are the ideas and behaviors which may be promulgated by the nursing student. PMID- 8718418 TI - Gemcitabine hydrochloride: combination of activity and tolerability. Proceedings of a symposium. Jerusalem, Israel, November 14, 1993. PMID- 8718419 TI - Preclinical characteristics of gemcitabine. AB - Gemcitabine (2',2'-difluorodeoxycytidine, dFdC) is a nucleoside analogue of deoxycytidine in which two fluorine atoms have been inserted into the deoxyribofuranosyl ring. Once inside the cell gemcitabine is rapidly phosphorylated by deoxycytidine kinase, the rate-limiting enzyme for the formation of the active metabolites gemcitabine diphosphate (dFdCDP) and gemcitabine triphosphate (dFdCTP). Gemcitabine diphosphate inhibits ribonucleotide reductase, which is responsible for producing the deoxynucleotides required for DNA synthesis and repair. The subsequent decrease in cellular deoxynucleotides (particularly dCTP) favours gemcitabine triphosphate in its competition with dCTP for incorporation into DNA. Reduction in cellular dCTP is an important self-potentiating mechanism resulting in increased gemcitabine nucleotide incorporation into DNA. Other self-potentiating mechanisms of gemcitabine include increased formation of active gemcitabine di- and triphosphates, and decreased elimination of gemcitabine nucleotides. After gemcitabine nucleotide is incorporated on the end of the elongating DNA strand, one more deoxynucleotide is added, and thereafter the DNA polymerases are unable to proceed. This action, termed "masked chain termination", appears to lock the drug into DNA because proof-reading exonucleases are unable to remove gemcitabine nucleotide from this penultimate position. Incorporation of gemcitabine triphosphate into DNA is strongly correlated with the inhibition of further DNA synthesis. Compared with ara-C, gemcitabine serves as a better transport substrate, is phosphorylated more efficiently, and is eliminated more slowly. These differences, together with self-potentiation, masked chain termination and the inhibition of ribonucleotide reductase, which are not seen with ara-C, may explain why gemcitabine is, and ara-C is not, active in solid tumours. This unique combination of metabolic properties and mechanistic characteristics suggests that gemcitabine is likely to be synergistic with other drugs that damage DNA, and also with other modalities such as radiation. PMID- 8718420 TI - Current chemotherapy of advanced non-small cell lung cancer. AB - Systemic chemotherapy for patients with a good performance status and advanced non-small cell lung cancer may result in prolonged survival and improved quality of life. However, few single agents have an activity of more than 15% and they have significant toxicity. Cisplatin is widely regarded as the single agent of choice. Compared with single-agent therapy, two-drug combinations generally provide higher response rates although survival benefit is marginal. Three-drug combinations generally provide no additional efficacy benefit and are associated with greater toxicity. A cisplatin-based combination with one other agent provides the best currently available therapeutic index. Chemotherapy may also improve the patient's quality of life. The toxicity of current chemotherapy is an important factor and there is a clear need for new cytotoxic agents with equivalent or greater activity yet a more acceptable toxicity profile. PMID- 8718421 TI - Phase II trials of single-agent activity of gemcitabine in patients with advanced non-small cell lung cancer: an overview. AB - A total of 361 patients have been entered into four phase II trials in which gemcitabine was given as a 30 min infusion in a schedule weekly x 3 q4W, at starting doses of 800-1250 mg/m2. Three of these trials produced response rates of 22.5%, 20% and 21.8% (all response rates were independently evaluated by an extramural Oncology Review Board). One small study of 30 evaluable patients produced a disappointing low response rate of 3%, but in this study the mean and median dose delivered was under 700 mg/m2. Gemcitabine was well tolerated with modest levels of traditional cytotoxicities such as myelosuppression, nausea and vomiting, and alopecia. Pooled data from two of the studies show improvement in a number of disease-related symptoms. Objective response rates by prognostic factor were determined: stage IIIa (30.5%), stage IIIb (18.8%), stage IV (19.5%); performance status 0 (31.8%), 1 (16.7%) and 2 (21.7%); female gender (23.5%), male (17.5%); age < 70 (18.6%), age > 70 (21.9%). Gemcitabine can be considered for use as a single agent in patients unable or unwilling to tolerate combination chemotherapy. Dose-response data suggest that a dose of 1000 mg/m2 or more is required for optimal therapeutic effect. The single-agent activity of gemcitabine together with its non-overlapping toxicity and novel mode of action suggest that this agent should also be investigated in combination with other active agents in non-small cell lung cancer. PMID- 8718422 TI - Safety profile of gemcitabine. AB - This paper reviews the toxicity profile of gemcitabine in a large group of patients (up to 790) from pivotal phase II studies, in which the drug was given intravenously as a 30 min infusion, in a schedule once a week for 3 weeks followed by a week of rest. The safety profile of gemcitabine is unusually mild for such an active agent in solid tumours. Haematological toxicity is mild and short-lived with modest WHO grades 3 and 4 for haemoglobin (6.4% and 0.9% of patients), leukocytes (8.1% and 0.5%), neutrophils (18.7% and 5.7%) and platelets (6.4% and 0.9%). The incidence of grade 3 and 4 infection associated with this level of myelosuppression was low (0.9% and 0.2%). Transaminase elevations occurred frequently, but they were usually mild, and rarely dose limiting. Mild proteinuria and haematuria were seen but were rarely clinically significant. There was no evidence of cumulative hepatic or renal toxicity. Nausea and vomiting was mild, rarely dose limiting, and generally well controlled with standard antiemetics. Flu-like symptoms were experienced in a small proportion of patients but were of short duration. Where oedema/peripheral oedema was experienced there was no evidence of any association with cardiac, hepatic or renal failure. Hair loss was rare, with WHO grade 3 alopecia reported in 0.5% of patients. There was no grade 4 alopecia. Furthermore, gemcitabine displayed minimal toxicity in elderly patients, and the side-effect profile does not seem to be affected by patient age. The adverse events typically experienced with cytotoxic agents, namely myelosuppression, nausea and vomiting and alopecia, are not seen to such a degree with gemcitabine, and this nonoverlapping toxicity profile suggests that gemcitabine is a promising agent for incorporation into combination chemotherapy regimens. PMID- 8718423 TI - Combination chemotherapy with gemcitabine and cisplatin in the treatment of advanced non-small cell lung cancer: preliminary results of an ongoing phase I/II study. AB - Collaborative phase I and II studies of the combination of gemcitabine and cisplatin in patients with advanced non-small cell lung cancer are ongoing at five centres in the UK and France. In the initial completed phase I study, 16 patients (15 evaluable) have been entered using a fixed dose of gemcitabine 1000 mg/m2 given as a 30 min intravenous infusion weekly for 3 weeks. On the third week the gemcitabine was immediately followed by cisplatin with pre- and post hydration. This regimen required only 1 night of hospitalization every 4 weeks. The study design was for sequential groups of patients to receive 3 dose levels of cisplatin (60 mg/m2, 75 mg/m2 and 100 mg/m2) but these doses would be modified and the number of patients at any dose level could be increased if significant toxicity was observed. Three patients were to be entered at the first two dose levels and 10 patients were to confirm the maximum tolerated dose (if reached) or expand the database on toxicity at the final predetermined dose level. The major haematological toxicities were neutropenia (grade 4 in 3 patients) and thrombocytopenia (grade 3 or 4 in 5 patients) but both were of short duration and uncomplicated. Grade 3 nausea and vomiting occurred in 12 patients but was no worse than would be expected from cisplatin alone. Alopecia was not a problem (no hair loss in 10 patients and grade 1 or 2 in 6 patients) and no significant renal or neurotoxicity was seen. A phase II study using cisplatin 100 mg/m2 in combination with gemcitabine 1000 mg/m2 has been opened and to date 19 patients are evaluable for response. Eight (42%) have achieved partial remissions. The study is ongoing and will recruit 50 evaluable patients. PMID- 8718424 TI - Symptomatic benefit from gemcitabine and other chemotherapy in advanced non-small cell lung cancer: changes in performance status and tumour-related symptoms. AB - Results from recent trials challenge the traditional view that chemotherapy offers no survival or quality of life benefits over best supportive care. Meta analyses of recent trials reveal a modest survival benefit for combination chemotherapy over best supportive care, although there is no strong evidence from randomized trials for superiority of combination over single-agent therapy. In chemotherapy trials where data on performance status change were collected, performance status improved in one-third of patients and remained constant in a further third. Fewer studies have measured changes in specific disease-related symptoms, but there are data from studies with gemcitabine which show improvements in a range of symptoms, including cough, haemoptysis, pain, dyspnoea and anorexia. Thus more patients benefit from chemotherapy than may be suggested by objective response. Surveys have shown that patients are more likely to accept intensive chemotherapy for what are perceived by health care professionals as potentially small benefits. Studies have shown evidence of cost savings associated with chemotherapy over best supportive care. PMID- 8718425 TI - Gemcitabine: clinical and economic impact in inoperable non-small cell lung cancer. AB - This study assesses retrospectively the clinical and economic impact of gemcitabine monotherapy on the management of inoperable stage III/IV non-small cell lung cancer in Germany. Based on current methods of clinical practice and using the best outcome data available, the costs and benefits of gemcitabine were compared to a dual therapy (ifosfamide/etoposide). While the two treatments showed broadly equivalent efficacy in terms of tumour response rate and survival, a cost analysis showed the potential for savings with gemcitabine. These largely related to hospital hotelling costs, due to the fact that gemcitabine may be given as an out-patient therapy. Further savings were found in investigative procedures and the management of treatment toxicity. Excluding the cost of the chemotherapy, gemcitabine was associated with potential savings of DM3,026 over two cycles of therapy, which included a 40% decrease in hospitalization costs and a 54% decrease in the cost of managing adverse events. We conclude that gemcitabine monotherapy could offer considerable cost savings while offering the potential for improved quality of palliative treatment compared to existing in patient treatments, and it may have a place in shifting care from an in-patient to an out-patient setting in line with recent health care reforms. PMID- 8718426 TI - Gemcitabine in advanced breast cancer. AB - The experience with single-agent gemcitabine in advanced or metastatic breast cancer is reviewed. In all studies, gemcitabine was administered as a 30 min intravenous infusion in cycles once a week for 3 weeks followed by 1 week of rest. In the first European study (gemcitabine 800 mg/m2/week), of 40 evaluable patients, 14 were chemo-naive, 7 had received adjuvant chemotherapy, and 19 had received chemotherapy for metastatic disease. There were 3 complete responders and 7 partial responders (all independently validated by an external Oncology Review Board) for an overall response rate of 25.0% (95% CI: 12.7%-41.2%). The median time to declaration of response was 1.9 months and the median duration of survival for all 40 efficacy-evaluable patients was 11.5 months. Haematological and non-haematological toxicities were particularly mild. WHO grade 3 and 4 toxicities included leukopenia (6.8% and 2.3% of patients), neutropenia (23.3% and 7.0%), AST (6.8% and 2.3%), ALT (18.2% and 0%), infection (0% and 2.3%), nausea and vomiting (25.0% and 2.3%), alopecia (2.3% and 0%). There was no grade 3 or 4 creatinine, proteinuria or haematuria. In the smaller US study (18 evaluable patients, all but one having received prior chemotherapy for stage IV disease) there were no responders. However, the mean dose delivered was very low (577 mg/m2/injection). In an ongoing European trial, with a starting dose of 1000 mg/m2, a number of partial responders have been seen in soft tissue, lung and liver. Gemcitabine's modest toxicity profile and single-agent activity make it an attractive candidate for trial in combination therapy in advanced breast cancer where treatment is currently given to palliate symptoms and improve quality of life. PMID- 8718427 TI - Phase II study of gemcitabine in previously platinum-treated ovarian cancer patients. AB - Fifty-one patients with histologically confirmed epithelial stage III or IV ovarian cancer were entered into a study in which gemcitabine 800 mg/m2 was given as a 30 min intravenous infusion in a cycle once a week for 3 weeks followed by a week of rest. Patients were aged 58 years (range 23-70 years) with WHO performance status 0-2, and had received up to two different chemotherapy regimens. Thirty-eight patients had received only one prior platinum-containing chemotherapy regimen whereas 9 had received a first-line regimen on more than one occasion. A further 3 patients had received two different regimens. Of 42 patients evaluable for response, 8 (19%; 95% CI: 9%-34%) were partial responders. Seven of the 8 responders were resistant to first-line platinum-based therapy. Median duration of response was 8.1 months (range 4.4-12.5 months). Median progression-free survival was 2.8 months (range 0.2-12.5 months). Haematological toxicity with gemcitabine was modest, with grade 3 leukopenia (11 patients) and grades 3 and 4 thrombocytopenia (6 patients). Grade 3 non-haematological toxicity included nausea/vomiting (6 patients) and elevated AST/ALT (1 patient), while dose-limiting non-haematologic toxicity consisted of flu-like symptoms (2 patients), peripheral oedema (1 patient) and lethargy (1 patient). The activity and modest haematological and non-haematological toxicity seen with gemcitabine suggest that this agent should be further evaluated in the treatment of patients with ovarian cancer and in combination chemotherapy regimens, primarily in combination with platinum. PMID- 8718428 TI - Sequential trends in overall and cause-specific mortality in diabetic and nondiabetic Pima Indians. AB - OBJECTIVE: To compare sequential trends in overall and cause-specific death rates for diabetic and nondiabetic Pima Indians. RESEARCH DESIGN AND METHODS: Underlying causes of death in Pimas aged > or = 15 years old were determined for the years 1975-1989 from review of death certificates and medical records. Overall and cause-specific death rates were compared for consecutive intervals. RESULTS: The all-causes death rate, age- and sex-adjusted, did not change significantly between the first and second halves of the study for diabetic (death rate ratio [DRR] = 0.99, 95% CI 0.70-1.4) or nondiabetic Pimas (DRR = 0.92, 95% CI 0.74-1.1). Among diabetic Pimas, however, the death rate for diabetic nephropathy declined from 2.7 to 1.5/1,000 person-years (DRR = 0.55, 95% CI 0.33-0.93), with ischemic heart disease (IHD) replacing diabetic nephropathy as the leading cause in the second half (DRR = 1.5, 95% CI 0.91-2.6). For diabetic and nondiabetic Pimas combined, the death rate in three consecutive 5 year periods declined progressively for alcoholic liver disease (P = 0.024) and external causes of death (P = 0.016), the largest component of which was automobile accidents. CONCLUSIONS: The decrease in death rate for diabetic nephropathy may be a result of greater access to and improvements in renal replacement therapy. Because of shared risk factors, however, the IHD death rate increased and largely offset the decrease in diabetic nephropathy deaths. The decline in deaths from alcoholic liver disease and from automobile accidents parallels the national trend. PMID- 8718429 TI - Acute effect of cigarette smoking on glucose tolerance and other cardiovascular risk factors. AB - OBJECTIVE: To investigate the acute effect of cigarette smoking on glucose tolerance, insulin sensitivity, serum lipids, blood pressure, and heart rate. RESEARCH DESIGN AND METHODS: This nonrandomized experimental control trial in a tertiary care center included 20 healthy chronic smokers and 20 age-, sex-, and BMI-matched healthy volunteers. Two oral glucose tolerance tests (OGTTs) were performed on each subject. Three cigarettes were smoked during the first 30 min in one of the tests. Serum glucose, insulin, and C-peptide levels were measured every 30 min; the area under the curve (AUC) and the insulin sensitivity index (ISI) were calculated; serum total cholesterol, LDL cholesterol, HDL cholesterol, and triglyceride levels were measured at 0 and 180 min; and blood pressure and heart rate were recorded every 5 min throughout 180 min. RESULTS: Smoking acutely impaired glucose tolerance: the AUC for glucose in smokers was 25.5 +/- 1.03 mmol/l (mean +/- SE) (95% CI 22.9-28) during the smoking OGTT and 21.8 +/- 0.85 mmol/l (CI 19.2-24.3) in the control OGTT (P < 0.01); in nonsmokers, it was 19.7 +/- 0.3 mmol/l (CI 18.8-20.5) in the smoking OGTT and 18.7 +/- 0.35 mmol/l (CI 17.8-19.5) in the control OGTT (P < 0.05). Smoking acutely increased serum insulin and C-peptide levels and decreased ISI only in smokers: ISI in smokers was 55 +/- 2.8 (CI 47.4-62.6) in the control OGTT and 43 +/- 2.7 (CI 35.4-50.6) in the smoking OGTT (P < 0.05). Smoking acutely caused a rise of serum total cholesterol levels in both groups and increased LDL cholesterol and triglyceride serum levels significantly only in smokers (P < 0.05). A significant rise of blood pressure and heart rate while smoking was present in all the subjects. CONCLUSIONS: Smoking acutely impaired glucose tolerance and insulin sensitivity, enhanced serum cholesterol and triglyceride levels, and raised blood pressure and heart rate. These findings support the pathogenetic role of cigarette smoking on cardiovascular risk factors. PMID- 8718430 TI - Deviation from developmentally appropriate self-care autonomy. Association with diabetes outcomes. AB - OBJECTIVE: Treatment of IDDM in youth emphasized balancing children's self-care autonomy with their psychological maturity. However, few data exist to guide clinicians or parents, and little is known about correlates of deviations from this ideal. RESEARCH DESIGN AND METHODS: In this cross-sectional study, IDDM self care autonomy of 100 youth was assessed using two well-validated measures. Three measures of psychological maturity (cognitive function, social-cognitive development, and academic achievement) were also collected for each child. Composite indexes of self-care autonomy and of psychological maturity were formed, and the ratio of the self-care autonomy index to the psychological maturity index quantified each child's deviation from developmentally appropriate IDDM self-care autonomy. Based on these scores, participants were categorized as exhibiting constrained (lower tertile), appropriate (middle tertile), or excessive (higher tertile) self-care autonomy. Between-group differences in treatment adherence, diabetes knowledge, glycemic control, and hospitalization rates were explored. RESULTS: Analysis of covariance controlling for age revealed that the excessive self-care autonomy group demonstrated less favorable treatment adherence, diabetes knowledge, hospitalization rates, and, marginally, glycemic control. Excessive self-care autonomy increased with age and was less common among intact two-parent families but was unrelated to other demographic factors. CONCLUSIONS: The findings indicate caution about encouragement of maximal self care autonomy among youth with IDDM and suggest that families who succeed in maintaining parental involvement in diabetes management may have better outcomes. PMID- 8718431 TI - Antibodies to bovine serum albumin in Brazilian children and young adults with IDDM. AB - OBJECTIVE: To evaluate the prevalence of IgG antibodies to bovine serum albumin (BSA) in a cohort of Brazilian children and young adults with IDDM. RESEARCH DESIGN AND METHODS: Sera from 81 subjects with < 1 year of IDDM (group 1), III subjects with > 1 year of IDDM (group 2), and 207 normoglycemic subjects were tested using an immunofluorimetric assay. A receiver-operating-characteristic curve was used to establish the threshold of anti-BSA antibody titers defining the positivity of the assay. RESULTS: The distribution of the fluorimetric index (FI) of anti-BSA antibodies did not have a gaussian profile. Rank sum of FI was significantly higher in patients than in control subjects (P < 0.0001). Average logFI values of both IDDM groups were significantly higher than that of the control group (P < 0.005 for both groups). There was a trend toward higher FI levels in group 1 than in group 2 (P = 0.06). A FI cutoff of 0.7 optimized the ratio of true-positive to false-positive of the assay, with the best equilibrium between sensitivity and specificity. The prevalence of anti-BSA antibodies was 52% in group 1, 47% in group 2, and 28% in the control group (P = 0.0001). An independent association between anti-BSA antibodies and IDDM, with an odds ratio of 3.03 (P < 0.0001), was observed in a logistic regression analysis. However anti-BSA antibodies explained only 5% of the variability of IDDM versus NIDDM. CONCLUSIONS: Our results confirm that the prevalence of anti-BSA antibodies is higher in IDDM subjects than in control subjects, even after 1 year of diabetes. However, a large overlap of antibody titers is observed in patients and control subjects, suggesting that anti-BSA antibodies are neither sensitive nor specific markers of IDDM. PMID- 8718432 TI - Raised serum sialic acid concentration in NIDDM patients with and without diabetic nephropathy. AB - OBJECTIVE: Raised serum sialic acid concentration is a strong predictor of cardiovascular mortality in the general white population. A progressive increase in cardiovascular morbidity and mortality takes place in relation to increasing albuminuria in NIDDM patients. Therefore, we investigated the potential association between serum sialic acid and micro- and macroangiopathy in NIDDM patients. RESEARCH DESIGN AND METHODS: We studied a prevalence cohort of all white NIDDM patients < 76 years of age attending a diabetic clinic during 1 year. Of the patients, 319 had normoalbuminuria, 148 had microalbuminuria, and 75 had macroalbuminuria (diabetic nephropathy was in 47 of 75 patients); 66 nondiabetic age- and sex-matched subjects acted as a control group. Blood samples were taken for measurements of sialic acid, lipids, creatinine, and HbA1C. Retinopathy was assessed by funduscopy. The prevalence of cardiovascular disease was based on Minnesota-coded electrocardiograms and the World Health Organization cardiovascular questionnaire. RESULTS: A progressive raise in serum sialic acid was demonstrated with an increasing urinary albumin excretion rate: [median (range)] 2.02 (1.55-2.63); 2.42 (1.47-6.48); 2.67 (1.57-5.86), and 2.95 (1.52 7.86) mmol/l in nondiabetic subjects, NIDDM patients with normoalbuminuria, microalbuminuria, and diabetic nephropathy, respectively (P < 0.05 or less for differences between groups). Multiple linear regression analysis showed that serum cholesterol concentration, serum HDL cholesterol concentration, BMI, albuminuria, smoking, and cardiovascular disease correlate independently with logarithmic (10) serum sialic acid concentration. CONCLUSIONS: Our study revealed a progressive raise in serum sialic acid with increasing urinary albumin excretion rate in NIDDM patients. Furthermore, several modifiable cardiovascular risk factors were associated with serum sialic acid. PMID- 8718433 TI - Prevalence of self-reported erectile dysfunction in people with long-term IDDM. AB - OBJECTIVE: The purpose of this report is to examine the prevalence of erectile dysfunction and relationships to other characteristics in men with younger-onset diabetes. RESEARCH DESIGN AND METHODS: In a population-based cohort study in southern Wisconsin, prevalence of erectile dysfunction was measured based on self reports in men who were 21 years of age or older, were < 30 years of age at diagnosis of diabetes, had 10 or more years of diabetes, and were taking insulin (n = 365). RESULTS: Of the study group, 20% reported a history of erectile dysfunction. The prevalence of erectile dysfunction increased with increasing age (from 1.1% in those 21-30 years of age to 47.1% in those 43 years of age or older, P for trend < 0.0001) and with increasing duration of diabetes (P for trend < 0.0001). Erectile dysfunction was associated with presence of severe diabetic retinopathy, a history of peripheral neuropathy, amputation, cardiovascular disease, a higher glycosylated hemoglobin, use of antihypertensive medications, and higher BMI. CONCLUSIONS: These data suggest that tighter glycemic control and careful selection of antihypertensive medications might prove beneficial. PMID- 8718434 TI - Capillary blood on filter paper for determination of HbA1c by ion exchange chromatography. AB - OBJECTIVE: To facilitate HbA1c determination, we evaluated an HbA1c filter paper system enabling capillary blood sampling at home by the patients. RESEARCH DESIGN AND METHODS: Capillary blood (two drops) was applied to a filter paper (HbA1c Via Post) and sent to the laboratory where a small disc was punched out on the filter paper. Hemoglobin was eluted from the disc in a buffer containing cysteine to eliminate the interfering glutathione adduct (HbA3) formed during storage. Analysis was performed by ion-exchange chromatography (Mono S, high-performance liquid chromatography), and the eluate was compared with hemolysate of venous blood from 41 patients. The stability of blood impregnated on filter paper was checked at different temperatures over different periods of time. RESULTS: There was an excellent agreement (r = 0.99) between HbA1c values from capillary blood on filter paper and HbA1c values from venous blood. HbA1c values were constant when stored on filter paper for 5-7 days at 20-21 degrees C (room temperature) or at 4-6 degrees C (refrigerator) for 10 days as well as at -70 degrees C for several months after blood sampling. A new chromatographic-interfering hemoglobin fraction both from venous and capillary samples was identified as free alpha chain of hemoglobin. CONCLUSIONS: The HbA1c filter paper system enables capillary blood sampling at home, eliminates the need of vein puncture in children and adults, and provides the diabetologist with an HbA1c value when the patient visits the clinic without a need for a previsit phlebotomy. PMID- 8718435 TI - Proinsulin immunoreactivity in recent-onset IDDM: the significance of insulin antibodies and insulin autoantibodies. AB - OBJECTIVE: To study the natural history of fasting proinsulin immunoreactivity (PIM) during the first 30 months of IDDM and its relationship to fasting C peptide and insulin antibodies. RESEARCH DESIGN AND METHODS: An incidence cohort of 204 consecutive newly diagnosed IDDM patients were followed prospectively, having blood drawn for measurements at diagnosis and at 1, 3, 6, 9, 12, 18, 24, and 30 months. A sensitive enzyme-linked immunosorbent assay was used for the determination of PIM. RESULTS: All patients had detectable fasting PIM in plasma at diagnosis, with a median value and interquartile range of 3.5 pmol/l (2.2 6.2). The median PIM level increased during the first months of IDDM to reach a peak at 9-12 months (9.9-10.3 pmol/l). PIM then declined gradually to 5.6 pmol/l (1.9-13.5) at 30 months without reaching baseline. PIM at each time point was widely scattered in a skewed log-normal distribution without signs of bimodality. After the onset of insulin treatment, median insulin antibody level increased and declined in a similar pattern. Both PIM and antibody level were significantly higher in children and adolescents compared with adults. However, stepwise multiple regression analysis showed that age was only of minor importance for the PIM variation during the study period. Insulin antibody level and fasting C peptide were the major determinants at 3-30 months, accounting for approximately 40% of the variation (R2). Blood glucose was of minor importance, and insulin dose, HbA1c, and BMI were of no importance. The correlation between fasting PIM and fasting C-peptide improved (R2 doubled) if the insulin antibody level was accounted for. Further, the slope of the correlation curve between PIM and C peptide increased threefold when antibody binding was > 4%. At diagnosis, insulin autoantibodies could be detected in 19% of the patients. Their presence predicted higher proinsulin at 1-3 months, a higher insulin dose the 1st year, and higher levels of insulin antibodies later in the study. CONCLUSIONS: Circulating insulin antibodies may affect the level of PIM in IDDM, probably by adding a pool of IgG bound PIM thereby increasing half-life and plasma concentration. This may explain why C-peptide and PIM levels do not change in concert during the 1st years of IDDM. Unlike C-peptide, PIM can not therefore quantitate beta-cell secretion unless the presence of insulin antibodies is ruled out. PMID- 8718436 TI - Effects of troglitazone: a new hypoglycemic agent in patients with NIDDM poorly controlled by diet therapy. AB - OBJECTIVE: To investigate the clinical efficacy of troglitazone, a newly developed oral hypoglycemic agent, in patients with NIDDM. RESEARCH DESIGN AND METHODS: There were 284 NIDDM patients (20-82 years of age) whose glycemic control while on a diet was judged stable but was judged unsatisfactory (fasting plasma glucose [FPG] > or = 8.3 mmol/l) when entered into a multicenter and double-blind study with parallel groups study. They were randomly allocated into two groups, the troglitazone group (the T group: 400 mg/day p.o.) and the placebo group (the P group), and were treated with test drugs for 12 weeks. RESULTS: We evaluated efficacy in 136 patients of the T group and 126 patients of the P group. There was no significant difference in any of baseline characteristics between the T and P groups. In the T group, FPG and HbA1c decreased significantly after treatment (before versus after, FPG 10.1 +/- 1.6 vs. 8.8 +/- 1.9 mmol/l, P < 0.001; HbA1c: 8.6 +/- 1.5 vs 8.1 +/- 1.7%, P < 0.001). FPG and HbA1c did not change after treatment in the P group (before versus after, FPG 10.1 +/- 1.8 vs. 9.9 +/- 2.1 mmol/l; HbA1c 8.5 +/- 1.5 vs. 8.6 +/- 1.6%). Of 136 patients in the T group, 62 (45.6%) were classified as responders. Serum triglyceride level also decreased in the T group but not in the P group. Body weight increased slightly only in the T group. There were no differences in changes in blood pressure between the two groups. No serious adverse events occurred in either group. CONCLUSIONS: Troglitazone at 400 mg/day decreased FPG and HbA1c significantly in NIDDM patients who had failed to respond to diet therapy. Troglitazone, developed as a drug to enhance insulin action, can be a useful hypoglycemic agent for the treatment of NIDDM. PMID- 8718438 TI - The effect of metformin on adipose tissue metabolism and peripheral blood flow in subjects with NIDDM. AB - OBJECTIVE: To examine the effect of metformin on net lactate and glycerol release in NIDDM subjects, we used abdominal subcutaneous microdialysis combined with 133Xe clearance. Skeletal muscle blood flow (MBF) was assessed simultaneously both before and after metformin treatment. RESEARCH DESIGN AND METHODS: Nine male patients with NIDDM (age 53 +/- 2 years [mean +/- SE]; BMI 30.2 +/- 1.4 kg/m2; body fat 23.0 +/- 2.6 kg; diabetes duration 4.6 +/- 1.5 years; six of nine receiving sulfonylurea treatment) were recruited into an open study. They were studied after an overnight fast, both before and after 1 week of additional treatment with 500 mg metformin three times daily. Nine weight- and age-matched nondiabetic subjects served as a control group. RESULTS: Postabsorptive net subcutaneous lactate release increased (149 +/- 50 vs. 475 +/- 127 nmol.100 g 1.min-1, P < 0.05) whereas plasma lactate was unchanged after metformin treatment in the NIDDM patients. The net decrease of glycerol release 90 min after an oral glucose tolerance test was more pronounced (110 +/- 30 vs. 199 +/- 20 nmol.100 g 1.min-1, P < 0.05) after metformin treatment. Both adipose tissue blood flow (ATBF) (1.5 +/- 0.1 vs. 2.3 +/- 0.2 ml.100 g-1.min-1, P < 0.01) and MBF (3.2 +/- 0.4 vs. 4.2 +/- 0.5 ml.100 ml-1.min-1, P < 0.05) increased after metformin treatment. CONCLUSIONS: In this open study, postabsorptive net lactate release in abdominal subcutaneous adipose tissue was clearly increased in NIDDM patients after metformin treatment. Basal ATBF as well as MBF was improved after metformin treatment. Whether this reflects enhanced metabolic control or is a drug-specific effect remains to be established. PMID- 8718437 TI - Plasma fibrinogen in NIDDM: the Rotterdam study. AB - OBJECTIVE: To compare plasma fibrinogen levels across groups of subjects with and without NIDDM with respect to diabetes therapy and to evaluate the influence of metabolic control and other selected factors. RESEARCH DESIGN AND METHODS: In a cross-sectional study, plasma fibrinogen was measured in 2,971 elderly subjects aged 55 years and older (mean age 70.8 years). This was part of the baseline examination of the Rotterdam Study, a population-based study of chronic diseases in the elderly. RESULTS: Plasma fibrinogen levels did not differ among subjects with and without NIDDM (2.84 vs. 2.81 milligrams, P = 0.5). After adjustment for age, sex, smoking, BMI, and waist-to-hip ratio, the plasma fibrinogen levels were higher in subjects receiving insulin therapy. In those without diabetes (n = 2,640), with diabetes but not taking antidiabetes medication (n = 196), taking oral medication (n = 99), and taking insulin (n = 36), the age- and sex-adjusted fibrinogen levels were 2.82 (SE 0.01), 2.79 (0.05), 2.79 (0.07), and 3.23 (0.11) milligrams, respectively (P < 0.005). Adjustment for leukocyte count or serum creatinine level did not affect the observed association, while adjustment for fructosamine decreased the differences. CONCLUSIONS: Plasma fibrinogen levels are elevated in insulin-treated NIDDM patients, which may reflect worse metabolic control in this subgroup. PMID- 8718439 TI - The development of foot deformities and ulcers after great toe amputation in diabetes. AB - OBJECTIVE: Our aim was to compare the prevalence and severity of foot deformities and the development of ulcerations in patients after a great toe amputations. RESEARCH DESIGN AND METHODS: We evaluated the presence of deformities of the toes and metatarsophalangeal joints (MTPJs) in patients with a great toe amputation who had an intact unamputated contralateral foot. The contralateral foot served as the patient's own control. We used a binomial test for paired data to compare the presence of deformity and ulcer formation and Fisher's exact test to compare joint flexibility in toes and MTPJs with foot deformities. RESULTS: There were more deformities of the second (P = 0.012) and third (P = 0.002) toes and lesser MTPJs (P < 0.05) and more rigid deformities of the second (P = 0.002) and third (P = 0.016) toes and second MTPJs (P = 0.035) in feet with great toe amputations. New ulcers were more common in feet that had an amputation (P = 0.002). CONCLUSIONS: We concluded that amputation of the great toe contributes to the development of deformities of the second and third toes and lesser MTPJs and new ulcer formation in patients with diabetes. When deformities were present, the second and third toes and second MTPJ were more severe in feet with a great toe amputation. PMID- 8718440 TI - Efficacy of feedback from quarterly laboratory comparison in maintaining quality of a hospital capillary blood glucose monitoring program. AB - OBJECTIVE: A 1-year randomized prospective study was conducted to investigate the efficacy of feedback from split-sample testing as part of a capillary blood glucose quality assurance program. RESEARCH DESIGN AND METHODS: A total of 124 nurses were randomized to either group A (quarterly comparisons with feedback) or group B (no feedback). The measure of nurse accuracy against the laboratory at 0, 6, and 12 months was determined by an additional five to seven split-sample tests without giving feedback to either group. Mean accuracy was determined in terms of percent absolute deviation from the laboratory result and a clinical consensus limit of +/- 20% deviation from the laboratory. RESULTS: By 12 months, there was a significant effect of feedback on nurse agreement with the laboratory method (P = 0.022 when agreement was scored as the mean percent absolute difference and P = 0.002 when agreement was scored in terms of the +/- 20% clinical consensus limit). Nurses in the group who had received no quarterly feedback from split sample testing produced a 3.5% greater mean percent absolute deviation from the laboratory method and 12% fewer comparisons within the acceptable +/- 20% range. CONCLUSIONS: Feedback received from split-sample testing has a significant effect in maintaining accuracy in capillary blood glucose monitoring. PMID- 8718441 TI - Five-year prospective study of glomerular filtration rate and albumin excretion rate in normofiltering and hyperfiltering normoalbuminuric NIDDM patients. AB - OBJECTIVE: To evaluate the evolution of glomerular filtration rate (GFR) and albumin excretion rate (AER) of normofiltering (NF) and hyperfiltering (HF) normoalbuminuric NIDDM patients. RESEARCH DESIGN AND METHODS: A longitudinal study of 32 normoalbuminuric (AER < 20 micrograms/min) NIDDM patients and 20 age , sex-, and BMI-matched normal individuals was done. Subjects had their GFR (51Cr labeled EDTA single-injection method) measured at entry and after 40 and 60 months. At entry, 13 NIDDM patients had GFR values above the upper limit of the normal range in our laboratory (> 137 ml.min-1 x 1.73 m-2) and were considered as HF. In NIDDM patients, the 24-h AER (radioimmunoassay), HbA1c, urinary urea, and mean arterial blood pressure (MBP) were analyzed at entry and after 40 and 60 months. RESULTS: There was a significant decline of GFR in NIDDM patients and normal subjects at 60 months. The decline was significantly greater in HF patients (-0.61 ml.min-1.month-1; P = 0.001) than in NF (-0, 18) and control subjects (-0, 14); the rate of change in NF and control subjects was the same (P > 0.05). In stepwise multiple regression analysis, with GFR decline as the dependent variable and GFR and AER at baseline, age and change in MBP, change in urinary urea, change in HbA1c, and change in therapy as independent variables, only baseline GFR (R2 = 0.19, P = 0.002) and age (R2 = 0.31, P = 0.048) were significantly related to the outcome. At 60 months, AER raised > 20 micrograms/min in three HF and in four NF patients. In logistic regression analysis, only higher initial AER (although still in the normal range; P = 0.037) and an increase in urinary urea (P = 0.021) were significantly related to the later development of microalbuminuria. CONCLUSIONS: The GFR of normoalbuminuric NIDDM patients declines significantly over 60 months. This decline is associated to baseline GFR and age. HF NIDDM patients show a faster decline in GFR than NF patients, whose GFR falls at a rate that is compatible with the age-related change observed in normal control subjects. The development of microalbuminuria is related to higher baseline AER and to increases in urinary urea and is similar in NF (4 of 19) and HF (3 of 13) NIDDM patients (P > 0.05). PMID- 8718442 TI - Referring patients with diabetes and vision loss for rehabilitation: who is responsible? PMID- 8718444 TI - Clinic variations hold important clues to the understanding and implementation of the DCCT results. PMID- 8718443 TI - Slowly progressive IDDM and malnutrition-related diabetes. PMID- 8718445 TI - Regarding ADA presidential address. PMID- 8718446 TI - Frequency of hypoglycemic episodes during intensive therapy with human insulin. PMID- 8718447 TI - U.K. prospective diabetes study. PMID- 8718448 TI - Threshold of HbA1c for the effect of hyperglycemia on the risk of diabetic microangiopathy. PMID- 8718449 TI - Latex allergy in diabetic patients: a call for latex-free insulin tops. PMID- 8718450 TI - IDDM and pancreatic carcinoma in Sardinia. PMID- 8718451 TI - Low incidence of false-positive exercise thallium 201 scintigraphy in a diabetic population. PMID- 8718452 TI - Cardiovascular disease and diabetes: issues raised at The European Association for the Study of Diabetes annual meeting. PMID- 8718453 TI - Metabolic regulation: a control analytic perspective. AB - A possible basis for a quantitative theory of metabolic regulation is outlined. Regulation is defined here as the alteration of reaction properties to augment or counteract the mass-action trend in a network reactions. In living systems the enzymes that catalyze these reactions are the "handles" through which such alteration is effected. It is shown how the elasticity coefficients of an enzyme catalyzed reaction with respect to substrates and products are the sum of a mass action term and a regulatory kinetic term; these coefficients therefore distinguish between mass-action effects and regulatory effects and are recognized as the key to quantifying regulation. As elasticity coefficients are also basic ingredients of metabolic control analysis, it is possible to relate regulation to such concepts as control, signalling, stability, and homeostasis. The need for care in the choice of relative or absolute changes when considering questions of metabolic regulation is stressed. Although the concepts are illustrated in terms of a simple coupled reaction system, they apply equally to more complex systems. When such systems are divided into reaction blocks, co-response coefficients can be used to measure the elasticities of these blocks. PMID- 8718454 TI - Elusive control. AB - The concept of a single rate-limiting step was proven to be too simplistic for understanding control and regulation of metabolism. Consequently, searches have identified relatively few steps with high control. Here we review a number of such searches and indicate what mechanisms may be responsible for this elusiveness of control. It turns out that this elusiveness of control has itself led to increased understanding of the roles played in metabolic control and regulation of such diverse factors as distributiveness of control, condition dependence, enzyme elasticity, homeostasis, control hierarchies, the input into a pathway, coenzyme sequestration, and redundancy and diversity of control function. PMID- 8718455 TI - Regulation of oxidative phosphorylation: the flexible respiratory network of Paracoccus denitrificans. AB - Paracoccus denitrificans is a facultative anaerobic bacterium that has the capacity to adjust its metabolic infrastructure, quantitatively and/or qualitatively, to the prevailing growth condition. In this bacterium the relative activity of distinct catabolic pathways is subject to a hierarchical control. In the presence of oxygen the aerobic respiration, the most efficient way of electron transfer-linked phosphorylation, has priority. At high oxygen tensions P. denitrificans synthesizes an oxidase with a relatively low affinity for oxygen, whereas under oxygen limitation a high-affinity oxidase appears specifically induced. During anaerobiosis, the pathways with lower free energy transducing efficiency are induced. In the presence of nitrate, the expression of a number of dehydrogenases ensures the continuation of oxidative phosphorylation via denitrification. After identification of the structural components that are involved in both the aerobic and the anaerobic respiratory networks of P. denitrificans, the intriguing next challenge is to get insight in its regulation. Two transcription regulators have recently been demonstrated to be involved in the expression of a number of aerobic and/or anaerobic respiratory complexes in P. denitrificans. Understanding of the regulation machinery is beginning to emerge and promises much excitement in discovery. PMID- 8718457 TI - Decoupling of the bc1 complex in S. cerevisiae; point mutations affecting the cytochrome b gene bring new information about the structural aspect of the proton translocation. AB - Four mutations in the mitochondrial cytochrome b of S. cerevisiae have been characterized with respect to growth capacities, catalytic properties, ATP/2e- ratio, and transmembrane potential. The respiratory-deficient mutant G137E and the three pseudo-wild type revertants E137 + I147F, E137 + C133S, and E137 + N256K were described previously (Tron and Lemesle-Meunier, 1990; Di Rago et al., 1990a). The mutant G137E is unable to grow on respiratory substrates but its electron transfer activity is partly conserved and totally inhibited by antimycin A. The secondary mutations restore the respiratory growth at variable degree, with a phosphorylation efficiency of 12-42% as regards the parental wild type strain, and result in a slight increase in the various electron transfer activities at the level of the whole respiratory chain. The catalytic efficiency for ubiquinol was slightly (G137E) or not affected (E137 + I147F, E137 + C133S, and E137 + N256K) in these mutants. Mutation G137E induces a decrease in the ATP/2e- ratio (50% of the W.T. value) and transmembrane potential (60% of the W.T. value) at the bc1 level, whereas the energetic capacity of the cytochrome oxidase is conserved. Secondary mutations I147F, C133S, and N256K partly restore the ATP/2e- ratio and the transmembrane potential at the bc1 complex level. The results suggest that a partial decoupling of the bc1 complex is induced by the cytochrome b point mutation G137E. In the framework of the protonmotive Q cycle, this decoupling can be explained by the existence of a proton wire connecting centers P and N in the wild type bc1 complex which may be amplified or uncovered by the G137E mutation when the bc1 complex is functioning. PMID- 8718456 TI - Regulation and control of compartmentalized glycolysis in bloodstream form Trypanosoma brucei. AB - Unlike other eukaryotic cells, trypanosomes possess a compartmentalized glycolytic pathway. The conversion of glucose into 3-phosphoglycerate takes place in specialized peroxisomes, called glycosomes. Further conversion of this intermediate into pyruvate occurs in the cytosol. Due to this compartmentation, many regulatory mechanisms operating in other cell types cannot work in trypanosomes. This is reflected by the insensitivity of the glycosomal enzymes to compounds that act as activity regulators in other cell types. Several speculations have been raised about the function of compartmentation of glycolysis in trypanosomes. We calculate that even in a noncompartmentalized trypanosome the flux through glycolysis should not be limited by diffusion. Therefore, the sequestration of glycolytic enzymes in an organelle may not serve to overcome a diffusion limitation. We also search the available data for a possible relation between compartmentation and the distribution of control of the glycolytic flux among the glycolytic enzymes. Under physiological conditions, the rate of glycolytic ATP production in the bloodstream form of the parasite is possibly controlled by the oxygen tension, but not by the glucose concentration. Within the framework of Metabolic Control Analysis, we discuss evidence that glucose transport, although it does not qualify as the sole rate-limiting step, does have a high flux control coefficient. This, however, does not distinguish trypanosomes from other eukaryotic cell types without glycosomes. PMID- 8718458 TI - Bartonella infections. PMID- 8718459 TI - Cerebrospinal fluid shunt infections. PMID- 8718460 TI - Pneumococcal infections in an era of multiple antibiotic resistance. PMID- 8718462 TI - Congenital cytomegalovirus infection and disease. PMID- 8718461 TI - Foodborne and waterborne illness in children. PMID- 8718463 TI - Recent advances in the prevention of pneumocystis carinii pneumonia. PMID- 8718464 TI - Invasive fungal infections in children: recent advances in diagnosis and treatment. PMID- 8718465 TI - Nematode infections in children. PMID- 8718466 TI - Hepatitis A and B vaccines in children. PMID- 8718467 TI - Adenoviral infections in children. PMID- 8718468 TI - Helicobacter pylori in infants and children. PMID- 8718469 TI - Urinary tract infections in infants and children. PMID- 8718470 TI - Diagnosis and treatment of congenital toxoplasmosis. PMID- 8718471 TI - Ambulatory phlebectomy compared to sclerotherapy for varicose and telangiectatic veins: indications and complications. PMID- 8718472 TI - Liposuction. PMID- 8718473 TI - Cosmetic laser surgery. AB - The fact that multiple wavelengths of light (and, therefore, different lasers) are needed to treat multicolored tattoos and several laser sessions are required for tattoo eradication in most instances, elimination of tattoos remains an expensive endeavor in terms of money and time. The obvious benefit of lasers in the treatment of tattoos, however, lies in their ability to eliminate tattoos without scarring or dyspigmentation. While no single commercially available laser system currently exists that can treat all tattoos, combination laser systems may make treatment of a wide range of tattoo colors a reality in the near future. PMID- 8718474 TI - What's new in sexually transmitted diseases. PMID- 8718475 TI - Newer antibiotics: a dermatologist's guide. PMID- 8718476 TI - Subcutaneous emphysema. AB - A medical emergency, the detection of subcutaneous emphysema requires thorough evaluation to exclude the multitude of disease processes that may demonstrate this clinical finding. Gas gangrene must be considered in the differential diagnosis of all forms of subcutaneous emphysema and infections with some species, such as C. novyi, may not produce gas at all. Isolation of C. septicum from the blood is almost always associated with colon cancer or hematologic malignancies. Nonclostridial gas gangrene in diabetic patients is indistinguishable clinically from clostridial gas gangrene. A unique and true dermatologic emergency is the detection of nontraumatic subcutaneous emphysema of the thigh with or without associated erythema, tenderness, or bullous lesions. This finding is associated with perforated viscus in a retroperitoneal location. Infections with gas-producing organisms continue to be a source of significant morbidity in modern times. PMID- 8718478 TI - Neurofibromatosis. PMID- 8718477 TI - Advances in viral infections. PMID- 8718479 TI - Molecular diagnosis of inherited skin diseases: the paradigm of dystrophic epidermolysis bullosa. PMID- 8718480 TI - Paraneoplastic dermatopathology: cutaneous paraneoplastic syndromes. PMID- 8718481 TI - Advances in the histologic diagnosis of cutaneous T-cell lymphoma. PMID- 8718482 TI - Update on the diagnosis of lichenoid dermatitis. PMID- 8718483 TI - Pemphigus: autoimmunity to epidermal cell adhesion molecules. PMID- 8718484 TI - Papillomavirus vaccines: current status and future prospects. PMID- 8718485 TI - Gene therapy for skin diseases. PMID- 8718486 TI - Identification performance by right- and left-handed listeners on dichotic CV materials. AB - Normative data from 24 right-handed and 24 left-handed subjects were obtained for the dichotic consonant-vowel (CV) materials (pa, ta, ka, ba, da, and ga) recorded on the Veterans Affairs (VA) compact disc (CD) Tonal and Speech Materials for Auditory Perceptual Assessment, Disc 1.0. Identification performance by the two subject groups was significantly different. With the right-handed subjects, identification performances on the materials presented to the right ear (RE) were better than performances on the materials presented to the left ear (LE) by (1) 16.3 percent for simultaneous onsets (RE = 72.8%; LE = 56.5%), (2) 11.3 percent for the 90-msec right-ear lag (RE = 86.3%; LE = 75.0%), and (3) 8.7 percent for the 90-msec left-ear lag condition (RE = 85.8%; LE = 77.2%). In comparison to the right-handed subjects, the left-handed subjects exhibited a smaller right-ear advantage and more inter-subject variability. With the left-handed subjects, identification performances on the materials presented to the right ear also were better than performances on the materials presented to the left ear by (1) 1.7 percent for simultaneous onsets (RE = 62.9%; LE = 61.1%), (2) 2.8 percent for the 90-msec right-ear lag (RE = 75.0%; LE = 72.2%), and (3) 3.1 percent for the 90 msec left-ear lag condition (RE = 75.9%; LE = 72.9%). The results are similar to previous dichotic CV data and indicate that the CVs recorded on the VA-CD provide valid estimates of identification performance on the dichotic CV materials. PMID- 8718487 TI - ABR slow wave and stimulus duration. AB - The slow wave (SW) component of the auditory brainstem response (ABR) was recorded in eight young adult subjects of both genders who had normal hearing sensitivity and who exhibited normal behavioral temporal integration (TI) functions. Test stimuli were 500- and 2000-Hz tone bursts, with rise and decay ramps of two periods, ranging in total duration from 2.5 msec to 44 msec. The responses appeared to be made up of contributions from both SW and wave V of the ABR. These composite waveforms showed a progressive, systematic, and statistically significant increase in peak amplitude and latency as a function of stimulus duration at 2000 Hz, but not at 500 Hz. Some pros and cons of recording the SW component during ABR audiometry are discussed, along with other advantageous properties of ABR SW. Additional approaches that may lead to the enhancement of the detectability of short-latency auditory evoked responses are noted. PMID- 8718488 TI - Comparison of performance with a conventional and a two-channel hearing aid. AB - Twenty experienced binaural hearing aid users evaluated a two-channel behind-the ear instrument, with a low band that offered dynamic range compression and a high band that provided linear amplification. After a 5-week trial period, data from the Profile of Hearing Aid Performance (PHAP), the Speech Perception in Noise (SPIN) test, as well as 2-cm3 coupler measurements were compared for the two channel experimental device and the subjects' single-channel aids. Most subjects showed significant improvement on the SPIN test and on the PHAP with the two channel aid. Subjective comments were predominantly enthusiastic, and 17 subjects (85%) chose to exchange their present instruments for the experimental aid. After a total of 19 weeks experience with the two-channel hearing aid, 15 subjects were reevaluated; the mean scores for the SPIN test and for most of the PHAP subscales were unchanged from those observed at the 5-week assessment. PMID- 8718489 TI - Phase effects on the middle and late auditory evoked potentials. AB - A masking level difference (MLD) paradigm was used to investigate the presence of an electrophysiologic correlate of the psychoacoustic MLD in the middle and late auditory evoked potentials. In experiment 1, middle latency potentials were recorded in six normal-hearing young adults using vertex and temporal electrode montages. Tone bursts of 500 Hz presented in SoNo and Spi No conditions produced no threshold differences that were consistent with an MLD. In experiment 2, late latency potential (P2) thresholds to 500-Hz tone bursts under various phase conditions in noise were measured and compared to behavioral thresholds from the same stimuli. Ten subjects provided behavioral and P2 thresholds to SoNo, S pi No, So N pi S pi N pi, SmNo, and SmNm conditions. The rank order of behavioral and P2 thresholds and MLDs was consistent with previous literature on the behavioral MLD. Cortical contributions were confirmed as necessary for the production of the electrophysiologic MLD and, by analogy, the psychoacoustic MLD. PMID- 8718490 TI - Comparison of probe insertion methods on estimates of ear canal SPL. AB - Real-ear sound pressure levels (SPLs) were compared among three methods used for positioning a probe microphone in the ear canal. The probe insertion techniques included (1) an acoustic method that incorporates use of the quarter-wave anti resonance property of the ear to determine acoustically the location of the probe tube relative to the eardrum; (2) a constant insertion depth method (25 mm from the intratragal notch); and (3) the earmold +5-mm method, which places the probe 5 mm beyond the tip of the earmold, thereby avoiding problems associated with the transition region where sound exits from the bore of the earmold into the larger ear canal. Measurements were obtained at 32 test frequencies in 24 adults with normal middle ear impedance. Results indicated that the SPLs measured by the acoustic method were modestly higher than those measured by the other two methods. This result was most evident in subjects with long ear canals (> 25 mm) and at high test frequencies (3.0 to 6.3 kHz). PMID- 8718491 TI - Postural adjustments produced by moving visual (horizontal optokinetic) patterns. AB - The effects of horizontal optokinetic stimulation (HOKS) on postural sway were examined in 30 normal subjects aged 20 to 75 years. Fixed-platform posturography was evaluated with the eyes open and closed and during HOKS (20-100 degrees/sec) in the rightward and leftward directions. Forward-backward sway was greater in amplitude than lateral sway under all viewing conditions. Sway amplitude was greater with eyes closed than with eyes open. HOKS significantly enhanced sway amplitude in both directions; however, significant differences occurred only between velocity extremes. Compared to younger subjects, older subjects showed greater forward-backward sway with eyes closed and greater lateral sway during HOKS. HOKS produces a visual cue that conflicts with the other two sensory signals regulating posture (vestibular and proprioceptive systems). Such conflicting visual cues may contribute to serious postural instability and falls in the elderly. The addition of optokinetic stimuli to fixed-platform posturography may enhance its diagnostic value. PMID- 8718492 TI - Dichotic listening to speech: VA-CD data from elderly subjects. AB - The Department of Veterans Affairs (VA) compact disc (VA-CD) Tonal and Speech Materials for Auditory Perceptual Assessment, Disc 1.0 contains three dichotic speech tests. Two of them, namely, dichotic digits and dichotic synthetic sentences, are tasks that are easily and accurately performed by young normal listeners. The third test, dichotic nonsense syllables, is a more difficult task for normal listeners to perform well. Few data have been gathered from use of the VA-CD with subjects other than young adults with normal hearing sensitivity. This account is about 19 elderly patients with hearing loss in a VA long-term care facility and how they performed on the trio of dichotic tasks on the disc. The digits, sentences, and syllables were presented at 80 dB SPL in a dichotic format requiring two responses to each trial. Trials using syllables were also done at 90 dB SPL to quantify the effects of hearing loss. Digits were recognized better than sentences; both were recognized better than syllables, and this was true for both ears. Compared to results from young listeners, correct responsiveness for older people was most reduced on the syllable task, but performance on dichotic sentences was also reduced. The results corroborate previous reports that suggest that many aging auditory systems do not process dichotic nonsense syllables as well as younger ones. The results also suggest that the dichotic procedures on the VA-CD provide a useful continuum of difficulty for the clinician and investigator. PMID- 8718493 TI - Genetic susceptibility to nephropathy in insulin-dependent diabetes: from epidemiology to molecular genetics. PMID- 8718494 TI - The genetics of the NOD mouse. PMID- 8718495 TI - Pancreas transplantation: an update. PMID- 8718496 TI - Insulin regulation of muscle glucose metabolism: role of pre-receptorial mechanisms. PMID- 8718497 TI - The basis of direct and indirect calorimetry and their potentials. PMID- 8718498 TI - History and current state of pediatric oncology and hematology in the United Arab Emirates. AB - There are three oncology centers in the United Arab Emirates, but only one with a dedicated pediatric unit (Tawam Hospital). The unit is 12 years old and is staffed by two pediatric oncologists working with pediatric but not oncology nurses. It handles approximately two thirds of the pediatric cancer cases in the country. Approximately 60% of all cases are either acute leukemia or lymphoma. A large variety of treatment regimens were used in the past for the same diseases, making audit of the unit's work very difficult. In the last 7 years a policy has evolved whereby uniform management protocols are used. A database is in existence, although not yet complete, and diagnostic facilities are continuing to improve. Imaging facilities are modern, and cell surface phenotyping for leukemia is available. There are plans to establish a bone marrow transplant unit. PMID- 8718499 TI - Unusual distribution of childhood cancer in Namibia. AB - A survey of childhood cancer was undertaken in Namibia from 1983 to 1988 to record all tumors in children less than 15 years of age. The national incidence of childhood cancer in the republic of Namibia was 55.5 per million children. The overall incidence rate was 73.3 per million in urban and densely populated areas, and 44.4 per million in the rural areas. This difference was not statistically significant. The relatively high 75 per million overall incidence of tumors amongst the white population group was probably due to the generally higher socioeconomic status and concomitant good medical care of this ethnic group during the study period. The significantly higher overall incidence of tumors (109 per million) recorded in the Rehoboth ethnic group, however, could not be accounted for by socioeconomic status or better health care facilities and remains unexplained. The cause of the increased incidence of central nervous system tumors in the Herero (26 per million) and osteosarcomas in the Kavango (11 per million) ethnic groups is also not clear and warrants further research. The apparent geographical cluster of tumors in northern and central Namibia was caused by the irregular distribution of the population. PMID- 8718500 TI - 3,4-dihydroxyphenylalanine (DOPA) metabolism in screening-detected and non screening-detected neuroblastoma. AB - To investigate the possible clinical application of the hypothesis that insufficient induction of 3,4-dihydroxyphenylalanine decarboxylase (DDC) causes accumulation and secretion of 3,4-dihydroxyphenylalanine (DOPA) in unfavorable neuroblastomas, we measured plasma DOPA in 28 neuroblastoma patients. Abnormally high levels were demonstrated in patients with neuroblastoma, and the levels in patients with clinical manifestations (median, 44,800 pg/ml; range, 17,700 to 220,000 pg/ml; n = 6) were significantly higher than those in patients detected by screening (median, 5825 pg/ml; range 2890 to 33,300 pg/ml; n = 22) (P = 0.0004). The catecholamine secretion profiles of the two groups were different, and it was suggested that the relative deficiency of DDC caused DOPA secretion in patients in the former group, whose prognosis was unfavorable, except in one case. In both groups, serial determination of plasma DOPA was a good monitor of the disease course. The higher plasma DOPA level (>9400 pg/ml) was significantly correlated with the patients' age (>1 year old) (P = 0.019), tumor stage (III, IV) (P = 0.029), and DNA diploidy (P = 0.018). These results are consistent with previous studies that demonstrated plasma DOPA was a useful marker in the diagnosis and follow-up of neuroblastoma. The results also indicate that higher plasma DOPA levels are associated with the unfavorable characteristics of neuroblastoma, which seem to support the hypothesis on DDC deficiency in unfavorable neuroblastoma. PMID- 8718501 TI - Childhood leukemia and statistical characteristics in 0- to 18-year-old patients in Hungary, diagnosed between 1988 and 1992. AB - A population-based cancer registry for childhood leukemia was started in Hungary in 1971. Data processing and analysis have been done at the Second Department of Pediatrics, Semmelweis University Medical School in Budapest, which is the main center for the treatment of childhood malignancies in Hungary. In 1992 a new computerized database structure was created in collaboration with Kansas University Medical Center, Kansas City, Kansas, United States. This work presents childhood leukemia frequency distribution and treatment results between 1988 and 1992 in Hungary. The number of patients diagnosed with leukemia under 18 years of age fluctuates between 69 and 82 cases per year. We present the main causes of death and outline our future objectives to improve the survival rate and quality of life. PMID- 8718502 TI - Early toxicity of intensified conditioning with etoposide combined with total body irradiation/cyclophosphamide or busulfan/cyclophosphamide in children undergoing autologous or allogeneic bone marrow transplantation. AB - In recent years, high dose chemotherapy followed by bone marrow rescue has been established as a common treatment of hematologic and solid tumor malignancies. Despite unequivocal success, relapse after transplant remains a serious problem, being the main cause of treatment failure. In an attempt to reduce relapse rates, we intensified the conditioning regimens consisting of busulfan/cyclophosphamide versus fractionated total body irradiation (f-TBI)/ cyclophosphamide by the addition of high dose etoposide. Toxicity profiles of 25 pediatric patients with hematologic malignancies undergoing intensified conditioning did not differ significantly between the two groups, except for a higher incidence of veno occlusive disease in busulfan-treated patients (3 of 13 patients) compared with the TBI group (0 of 12 patients). We observed no transplant-related mortality in neither group. Regimen-associated morbidity was moderate and reversible in all cases. Five patients died in each treatment arm, due to relapse of the underlying disease. We conclude that both regimens are feasible in marrow transplantation of pediatric patients. Open randomized trials are needed to assess the efficacy of intensified conditioning in terms of disease-free survival. PMID- 8718503 TI - Interleukin-1 production following T-cell-depleted and unmodified marrow grafts. AB - Interleukin-1 (IL-1) production by endotoxin-stimulated cultured monocytes from 31 participants in grafts of marrow depleted of mature cellular elements by treatment with soybean agglutinin and sheep red blood cells (SBA-E-) and 12 recipients of unfractionated bone marrow were studied and compared with normal controls. Patients were studied prior to marrow transplant (BMT) and at 1 month, 2 to 4 months, and 5 to 6 months post-transplant. Deficiencies in IL-1 production (<50 units) were detected in both transplant groups prior to and at 1 month post BMT. From 2 to 4 months post-transplant, 67% of the recipients of unmodified marrow and 45% of the recipients of SBA-E- marrow grafts produced a normal level of IL-1. By 5 to 6 months post-transplant and thereafter, the proportions of patients exhibiting deficiencies in IL-1 production in each group were equally low. We also evaluated the impact of early deficiencies of IL-1 on engraftment, hematopoietic function, and immunological reconstitution. Deficiencies in IL-1 production persisting to 2 to 4 months post-BMT did not significantly affect the degree of chimerism or the time to recovery of neutrophil counts to 500/mu l in recipients of either unmodified or T-cell-depleted marrow. Platelet recovery during the first 50 days post-transplant was significantly slower in the IL-1 deficient group, but thereafter rebounded, so that by 4 months post-BMT patients with initial deficiencies in IL-1 production achieved levels comparable with those attained by patients with normal production of IL-1. When we looked at the lymphocyte response to phytohemagglutinin (PHA), there was no difference detected among patients with or without IL-1 deficiency receiving unmodified transplants. In contrast, recipients of T-cell-depleted grafts exhibiting a prolonged deficiency of IL-1 experienced a slower rate of recovery of PHA responses. Our results suggest that IL-1 may play an important role in the early expansion of megakaryocytic precursors following an allogeneic marrow transplant and may facilitate the functional development of allogeneic lymphoid progenitors following a T-cell-depleted marrow graft. PMID- 8718504 TI - Chemotherapy with the "8 in 1" protocol for malignant brain tumors in children: a population-based study in Finland. AB - We evaluated the outcome of 68 children with malignant brain tumors treated with the "8 in 1" chemotherapy protocol in Finland from 1986 to 1993, comparing 5-year survival rates with those for a historical control group (from 1975 to 1985). For all malignant brain tumors, overall survival was 43% (vs 28% in the control group; P <0.05), and progression-free survival (PFS) was 43% (vs 23%; P <0.05). For medulloblastoma and primitive neuroectodermal tumor, survival was 63% (vs 35%; P <0.05), and the corresponding PFS was 59% (vs 35%; P = 0.15). For high grade glioma, both the survival rate and the PFS were 27% (vs 17%; P = NS). Thus the outcome was significantly better for our "8 in 1" -treated patients than for the historical controls, especially among the children with primitive neuroectodermal tumor and medulloblastoma. In contrast, those with high-grade gliomas and brain stem tumors seem to have received little benefit; different, more effective treatments are needed for these patients. PMID- 8718505 TI - Hydrops fetalis and neonatal leukemia in Down syndrome. AB - Four newborn infants with Down syndrome and manifestations of neonatal leukemia are described. One of the four was stillborn, two died shortly after birth, and a fourth survived and all evidence of leukemia disappeared in the first month of life. Three of the four cases had hydrops fetalis, and a fourth was a macerated stillborn. Nine other similar reported cases are reviewed. We conclude that neonatal leukemia in Down syndrome is a form of leukemia that is usually transient, with spontaneous recovery, but may be fatal at or around the time of birth with manifestations of hydrops fetalis, hepatosplenomegaly, and/or progressive liver disease. PMID- 8718506 TI - Hypertransfusion for spinal cord compression secondary to extramedullary hematopoiesis. AB - A 19-year-old girl with thalassemia intermedia presented with signs of thoracic spinal cord compression secondary to extramedullary hematopoiesis. She was started on a transfusion regimen to maintain a hemoglobin level of more than 12.5 g/dL. Clinical signs disappeared within the first week and circulating erythroblastemia was completely suppressed by the second week. Magnetic resonance imaging 4 weeks after diagnosis revealed near-complete resolution of the extradural mass, followed by gradual improvement in the posterior tibial somatosensory evoked potentials. Transfusion therapy may be diagnostically and therapeutically useful in spinal cord compression secondary to extramedullary hematopoiesis, obviating the need for surgery or radiotherapy. PMID- 8718507 TI - Intra-abdominal desmoplastic small round cell tumor in children: a clinicopathologic study. AB - Desmoplastic small round cell tumor (RCT) is a relatively newly recognized neoplasm. It has a very distinct morphologic and high-grade polyphenotypic expression with a very poor prognosis. It is commonly seen in adolescent boys. We describe two cases of intra-abdominal desmoplastic small RCT in young girls (5 and 11 years of age). In both cases, the exact origin of tumor in the abdomen could not be established. Histopathologic examination of the biopsy specimens showed distinct desmoplastic stroma and coexpression of various epithelial, neural, and muscle markers. These two patients were treated primarily by debulking surgery followed by adjuvant chemotherapy (RCT II protocol). One of the two had a stable disease while the other had a progressive disease while on treatment. Thus our findings support the diverse histogenesis of this tumor and its poor prognosis. PMID- 8718508 TI - Increased prolactin level and pituitary adenoma as a cause of headache in two patients with sickle cell disease. AB - In two patients with sickle cell disease who presented with headache pituitary adenoma and high levels of serum prolactin were found. Treatment with bromocriptine has controlled the headache. We believe that prolactin level determination should be part of the work-up of patients with sickle cell disease who complain of headache. PMID- 8718509 TI - Genetics and cancer: a second look. Introduction. PMID- 8718510 TI - Prospects for cancer genetics. AB - A number of distinct strategies have been used over the past two decades to uncover the genes involved in tumorigenesis. Their use raises the questions of whether we will require novel approaches in order to continue the progress of cancer genetics. Retrovirus mediated transduction of proto-oncogenes and transfection of tumour associated oncogenes are both inefficient ways of uncovering oncogenes, each being encumbered by technical obstacles that limit their utility. Improvements in the gene transfer strategy may yield a host of new oncogenes. New cloning techniques may have a role here as well as in revealing the existence of genes that are amplified in tumour cell genomes. The major technique for uncovering novel tumour suppressor genes--detection of loss of heterozygosity--is also limited in its sensitivity to detecting genes that are lost in large numbers of tumours. The techniques for uncovering these genes through analysis of pedigrees in which mutant versions of these genes are passed through the germline are encumbered by issues of penetrance and the complexities associated with the inheritance of polygenic traits. In both instances, improvements in data acquisition and analysis will reveal tumour suppressor genes that have proved elusive until now. Over the next decade, we will learn much about how the defects in another apparatus--that responsible for the maintenance of genomic integrity--contribute to cancer susceptibility. Beyond these genes lie yet others about which we seem to know little at present--those that induce the last stages of cancer including invasiveness and metastasis. These will represent an entirely new cohort of genes to be uncovered over the next decade. PMID- 8718511 TI - The genetic epidemiology of cancer. AB - Although every cancer is unique, similarities do exist with regard to the genetic epidemiology of many of the more common cancers such as breast, ovary, colon and prostate. For each of these cancers, a small subset of cases exist that are attributable to rare inherited mutant genes. Since cancer is a multistep process, individuals with inherited mutations are at increased risk and tend to develop cancer at an earlier age than do those who acquire the mutations after birth. The majority of cancer cases appear to be due to sporadic mutations that may occur as a result of spontaneous genetic events, environmental events or interaction between genetic and environmental factors. Genetic epidemiologists will continue to work towards the discovery of inherited cancer genes as well as to define the interaction of genotype with the environment. It is this research that scientists hope will be used to help in the identification of individuals with an increased probability of developing cancer as well as in the development of prevention and treatment strategies for individuals found to be at risk. PMID- 8718512 TI - Metabolic polymorphisms and cancer susceptibility. AB - The vast majority of cancers arise as a consequence of exposure to environmental agents that are toxic or mutagenic. In response to this, all higher organisms have evolved complex mechanisms by which they can protect themselves from environmental challenge. In many cases, this involves an adaptive response in which the levels of expression of enzymes active in the metabolism and detoxification of the foreign chemical are induced. The best characterized of these enzyme systems are the cytochrome P450s, the GSTs and the NATs. An unfortunate consequence of many of these reactions, however, is the creation of a toxic or mutagenic reaction product from chemicals that require metabolic activation before realizing their full carcinogenic potential. Altered expression of one or more of these drug metabolizing enzymes can therefore be predicted to have profound toxicological consequences. Genetic polymorphisms with well defined associated phenotypes have now been characterized in P450, GST and NAT genes. Indeed, many of these polymorphisms have been associated with decreased or increased metabolism of many tumour promoters and chemical carcinogens and hence offer protection against or increased susceptibility to many distinct tumour types. PMID- 8718513 TI - Genetics of embryonal tumours of childhood: retinoblastoma, Wilms' tumour and neuroblastoma. AB - There has been considerable progress in the past decade in elucidating the molecular basis of malignant transformation of retinoblastoma, Wilms' tumour and neuroblastoma. For retinoblastoma, the story is relatively simple and the laboratory focus should be on rapid detection of germline mutations, identifying additional genetic changes associated with tumour progression and the prevention of second primary cancers, if possible. In the case of Wilms' tumour, the locus (or loci) responsible for hereditary predisposition is still unknown. Furthermore, the WT2 locus has not been cloned and there is no consistent evidence of oncogene activation. Clearly, there is still much to be done before we fully understand the genetic basis of this disease. Finally, substantial insights have been gained from the molecular analysis of neuroblastomas, but this tumour is perhaps the least well understood. Two sites of allelic loss have been identified (1p36 and 14q32), but the presumptive suppressor genes that are the targets of these changes have yet to be identified. Furthermore, it is not clear if either of these loci is responsible for a genetic predisposition to develop this tumour. Although N-myc amplification is a powerful prognostic marker of aggressive tumours, no other oncogene has been shown to be activated in tumours lacking amplification. Finally, the NGFR pathway may have an important role in regulating differentiation and programmed cell death in these cells, but other NGFR family pathways or unrelated genes may be involved as well. Hopefully, the next decade will provide us with answers to many of these open questions. PMID- 8718514 TI - Germline mutations in the TP53 gene. AB - Since the majority of germline mutations in the TP53 gene seem to occur in LFS or LFL families, and these are rare, research is best conducted in a collaborative setting (Li and Fraumeni, in press). In a report from a meeting at Bethesda in 1993, the following areas were outlined for collaborative study: the correlation (if any) of phenotypes with specific mutation; age specific penetrance; cumulative cancer incidence; gender differences in tumour development in carriers; the effects of DNA damaging agents on individuals with a TP53 mutation; the frequency of TP53 germline mutations in cohorts of patients with rare childhood tumours (eg adrenocortical carcinoma); and the psychosocial aspects of predictive TP53 testing. In addition, if, as seems likely from recent data, X irradiation in these individuals induces DNA damage that is tolerated, urgent collaborative studies are needed to investigate new methods of screening, such as magnetic resonance imaging. Treatment modalities should be carefully chosen, and for this reason alone, predictive testing may be desirable in all LFS and LFL families. Individuals carrying TP53 mutations could be offered chemoprevention within trials in an effort to reduce their mortality from cancer. PMID- 8718515 TI - Nucleotide excision repair: variations associated with cancer development and speciation. AB - Nucleotide excision repair requires the action of multiple interacting proteins that locate damage in DNA, remove it as a short oligonucleotide and synthesize a replacement patch. Mutations in genes coding for these proteins give rise to a wide range of diseases involving skin carcinogenesis, neuronal decline and developmental disorders of bone and central nervous system. Complex clinical symptoms of more than one clinical disorder may occur because of mutations that influence protein-protein interactions. Significant differences in repair occur between individuals and species for which the molecular basis and phenotypic consequences have yet to be explained. PMID- 8718516 TI - Role of transgenic mice in identification and characterization of tumour suppressor genes. AB - Transgenic mice expressing various oncogenes have been generated and widely used to study their role in tumorigenesis. The development of targeted mutagenesis allows specific genes to be mutated in mouse embryonic stem cells, which can then be used to generate mice with a germline mutation in that gene. Embryonic stem cell technology has been used to reconstruct genetic lesions in tumour suppressor genes such as p53 and Rb1, originally identified from studies in humans. The tumour susceptibility phenotype of mutant mice has unveiled the tumour suppressor activity of specific genes that were not expected to have such a function. Transgenic and knock out mice will have an increasingly important role in the identification of novel tumour suppressor genes. PMID- 8718517 TI - Genomic imprinting and cancer. AB - Imprinting is vital for normal development, and disruption of imprinting mechanisms on syntenic chromosomes gives very similar phenotypes in mouse and humans. In addition, disruption of normal imprinting provides a plausible explanation for preferential LOH in some embryonal tumours. Moreover, there is evidence that in Wilms' tumour, dysregulation of specific imprinted genes may give rise to the cancer phenotype. Many more questions regarding genomic imprinting need to be answered before the associations described in this review can be properly understood. The most basic issues, such as when and how the imprint is established, can still only be speculated upon. Further study of new imprinted genes and the relationship between their domains and differential replication may show us higher control mechanisms than methylation alone. It remains to be seen if these epigenetic modifications are amenable to therapeutic change in the treatment of inherited syndromes and cancer, or if they can be used to assess individuals at risk of disease. Until then it is probably unwise to speculate on a single unifying theory that explains why a subset of the genome shows such a peculiar non-Mendelian form of inheritance. PMID- 8718518 TI - The genetics of programmed (apoptotic) cell death. AB - The genetic pathway for the activation and completion of programmed death of cells is as complex and well regulated as the pathway for cell proliferation. The identification of both the genetic elements in the signal transduction pathway involved with the initiation of programmed cell death, as well as the cellular machinery involved with DNA and cellular fragmentation into apoptotic bodies, is developing rapidly. Attempts at understanding how these elements are co-ordinated into a temporally discrete series of metabolic steps is only just beginning. This research not only will be fruitful from a basic science standpoint, but should also identify new approaches for cancer chemoprevention and therapy. PMID- 8718519 TI - Mutations of the RET proto-oncogene in multiple endocrine neoplasia type 2. PMID- 8718520 TI - The neurofibromatosis 2 (NF2) tumour suppressor gene: implications beyond the hereditary tumour syndrome? AB - The cloning of the gene that causes neurofibromatosis type 2 (NF2), a hereditary tumour syndrome typically associated with brain tumours such as vestibular schwannomas and meningiomas, represents another successful application of the "positional cloning" approach--that is, the isolation of a hereditary disease gene of unknown function, based on the determination of its chromosomal location in the human genome. The NF2 gene is homologous to a family of genes whose products, including moesin, ezrin, radixin and protein 4.1, appear to have an important role in bridging the cell membrane and the intracellular cytoskeletion. Mutation analyses have revealed that the NF2 tumour suppressor gene is frequently mutated not only in vestibular schwannomas and meningiomas from NF2 patients, but also in their sporadic counterparts, which represent approximately one third of all human brain tumours. Furthermore, malignant human tumours seemingly unrelated to the NF2 syndrome, such as malignant melanomas (derived from the neural crest) and malignant mesotheliomas (derived from pleural mesoderm), also frequently have mutations or deletions at the NF2 locus, suggesting a broader role of the NF2 gene in the initiation and progression of human neoplasms. PMID- 8718521 TI - Von Hippel-Lindau disease and sporadic renal cell carcinoma. AB - The VHL gene, isolated by positional cloning, encodes a protein of 284 aminoacids that has no homology with other proteins in the databases. The nucleotide sequence lacks domains that would suggest (a) a DNA binding protein, (b) nuclear localization, (c) enzymatic activity or (d) membrane localization. Studies are in progress on the size and cellular localization of the VHL protein and how it may function in growth regulation. How mutations in this small protein lead to a specific tumour spectrum presents an enormous research challenge. Germline mutations in the VHL gene are heterogeneous, and the resulting heterogeneity of mutations in the VHL protein that lead to disease suggests a protein whose function can be compromised by mutations over a large area. Study of the germline mutations and correlation with disease patterns provide the basis for a new clinical classification of von Hippel-Lindau disease. Somatic VHL mutations and hypermethylation of the VHL gene are found in some 75-80% of sporadic clear cell renal carcinomas. About 20% of clear cell renal carcinomas show neither VHL gene mutation or hypermethylation. Whether other chromosome 3 tumour suppressor genes have a pathogenetic role in clear cell renal carcinoma remains to be determined. Sorting out the contributions of different tumour suppressor genes to the pathogenesis of clear cell renal carcinomas will require assays demonstrating somatic mutation of candidate genes and functional assays to determine whether replacement of the mutant gene is associated with suppressed tumour growth. PMID- 8718522 TI - Genetics and malignant progression of human brain tumours. AB - Progression of gliomas to more malignant phenotypes involves numerous molecular genetic alterations. The genes affected by these alterations, the steps in malignant progression for which they are responsible, their normal function in controlling diverse cellular functions such as differentiation, signal transduction, cell cycle progression and angiogenesis and how they may act in concert with other tumour suppressor genes or oncogenes are some of the questions finally coming into focus and being studied. As other genes are discovered, their association with tumour progression can be assessed, coupled with current histopathology and used to determine more accurately patient prognosis and strategies for intervention. With the generation of specific reagents, such as monoclonal antibodies directed to glioma derived antigens or emerging gene therapy techniques designed to deliver toxic, antisense or reconstituting genes specifically to tumour tissue, new approaches will be devised that may finally be used to treat these tumours effectively. PMID- 8718523 TI - Recent advances in the molecular genetics of malignant melanoma. AB - The study of the molecular basis for sporadic and inherited melanoma has rapidly moved forward over the past several years. The crucial observation that chromosome 9p21 abnormalities occurred with high frequency in sporadic melanomas, coupled with the molecular demonstration of common 9p21 LOH, led investigators to focus on this region. Examinations of patterns of inherited susceptibility to melanoma established 9p21 as the site of the MLM locus. The localization of the CDK inhibitor CDKN2 to the region enabled the demonstration of its alteration in numerous sporadic solid tumours. Most importantly, the gene has been implicated in the pathogenesis of both inherited and sporadic melanoma. Much work needs to be done to further our understanding of the prevalence of CDKN2 mutations and the prognoses they confer. In addition, continued avenues of investigation are likely to involve further application of this approach to other regions of genomic instability in melanoma, especially chromosomes 1, 6 and 10. PMID- 8718524 TI - Molecular genetics of lung cancer. AB - The identification of genetic abnormalities in lung cancer and their consequences for cell growth, differentiation and function offer insights into normal cellular function and carcinogenesis. The integration of this knowledge with data from other fields, including epidemiology, toxicology, carcinogen metabolism, clinical genetics and clinical cancer therapy, promises advancements in lung cancer risk assessment, screening, prevention, prognostic assessment and therapy. PMID- 8718525 TI - Molecular genetics of sporadic and familial breast cancer. AB - Molecular genetic analysis of breast cancers indicates that the mechanisms underlying tumorigenesis are complicated. Many oncogenes and tumour suppressor genes have been implicated, encoding proteins that are important at many levels of cell regulation, from cell surface molecules responding to external signals (eg ERBB2) to nuclear factors controlling gene transcription (eg TP53, MYC). Several correlations have been found between certain genetic events and clinical outcome and have therefore proved useful prognostic indicators. The mapping and cloning of genes important in familial breast cancers (eg BRCA1) have provided the essential tools for pinpointing the genes that may be critical in early stage breast cancer as well as for developing genetic tests for predicting carrier status in breast cancer families. Clarification of the molecular consequences of mutation in breast cancer associated genes is beginning to address the factors that drive a normal breast cell to change into a breast cancer cell. However, these studies are still in their infancy, and considerable research will be required to complete the picture. PMID- 8718526 TI - Emergent issues in the genetics of intestinal neoplasia. AB - Mutation of the APC gene may be a common denominator of all human colon cancer- polypoid and non-polypoid familial cancer as well as sporadic occurrences. Fearon and Vogelstein (1990) have described a series of molecular changes during the progression of human colon cancer, beginning with mutations in APC. Min is a strain of the laboratory mouse carrying a nonsense mutation in Apc, the mouse homologue of APC. The Min strain has been used to test the effect of germline alterations in certain genes identified in the progression pathway of Fearon and Vogelstein. A deficiency in DNA cytosine methylase leads to a reduction in the tumour multiplicity of Min mice contrary to the a priori expectation based on the global hypomethylation of the DNA of early colonic neoplasms. Alterations in Kras had no perceptible effect on the tumour multiplicity of Min mice but may not have been successfully directed to the proliferative cell population. Constitutional mutation of p53 did not influence the multiplicity or histopathology of early Min induced intestinal tumours. The cause and effect analysis of the genetics of colon cancer is clearly in an early phase. An unlinked genetic factor interacting with Min in controlling intestinal tumour multiplicity is Mom1. A central goal for the near future is to identify the Mom1 gene product and to identify other loci that can interact with the Min mutation and affect tumour multiplicity or progression. Mouse chimaeras will permit an analysis of the clonality and cell autonomy of Min induced neoplasms and also of the action of Mom1. The results of these analyses will inform investigators as to what modes of prevention and therapy might be designed for particular tumour types. The Min strain thereby presents an opportunity to discover protective factors against human colon cancer. PMID- 8718527 TI - Molecular genetics of prostate cancer. AB - A number of genetic changes have been documented in prostate cancer, ranging from allelic loss to point mutations and changes in DNA methylation patterns. Up to now among the most consistent changes are those of allelic loss events, with the majority of tumours examined showing loss of alleles from at least one chromosomal arm. Chromosomes 8 and 13 appear to be the most frequently affected, with the former showing both loss of alleles from the short arm and gain of sequences on the long arm. Deletions of one copy of the RB gene are common, whereas deletion and/or point mutation of the TP53 gene is a less frequent event, at least in clinically localized tumours. Alterations in the E-cadherin/alpha catenin mediated cell-cell adhesion mechanism appear to be present in over one third of all prostate cancers and may be critical to the acquisition of metastatic potential of aggressive prostate cancers. In addition, altered DNA methylation patterns have been found in the majority of prostate cancers examined, suggesting an important role for methylation modulated gene expression in prostate carcinogenesis. Finally, the existence of prostate cancer susceptibility genes is suggested by study of familial clustering of prostate cancer, and it is expected that the identification of these genes will provide insight into critical rate limiting steps in the carcinogenic pathway of both inherited and sporadic disease. PMID- 8718528 TI - Ethical, social and counselling issues in hereditary cancer susceptibility. AB - Genetic testing for hereditary susceptibility to disease is new. Much has been learned from experience with Huntington's disease and other non-malignant conditions. There are some differences in the case of predisposition testing for cancer: there is often the perception that cancer is preventable and sometimes curable, in contrast to other hereditary conditions. Testing raises many issues new to the medical community and to the public as well. There is great concern that the explosive technology be used responsibly, so that the potential benefits of genetic knowledge are not eclipsed by the risks to autonomy, privacy and justice. Practical concerns about insurability and discrimination may inhibit some at risk individuals from taking advantage of this powerful technology. There has been considerable effort already in the UK, Europe and the USA at the research and social levels to create protection for individuals found to carry genetic susceptibility to disease. PMID- 8718529 TI - Row-to-row horizontal links may be associated with the transduction channels of hair cells. AB - In a geometric model of passive elastic elements in the hair bundles of vertebrate acousticolateralis hair cells, Geisler provides evidence against a role of horizontal links in sensory transduction. One premise of this model, the tenet that horizontal links are stretched in the same way as tip links, may be invalid. We propose that the horizontal links may be rigid, implying that the attachment points of the horizontal links in the stereociliary membrane must move when the hair bundle is deflected. Since the movement is linear with the angle of deflection, a connection of transduction channels and horizontal links is conceivable. PMID- 8718530 TI - Distribution of endothelin-1-like activity in the vestibule of normal guinea pigs. AB - Endothelin (ET) has been revealed to be a local hormonal regulator of pressure, fluid, ions and neurotransmitters. In order to investigate the mechanism of homeostasis within the microenvironment of the inner ear, the present study has examined the distribution of ET in the vestibule of normal guinea pigs, by immunohistochemistry using mouse antihuman ET IgG1 monoclonal antibody. ET-like activity was identified in the sensory epithelial cells, supporting cells, dark cells, transitional cells, vestibular membrane, semicircular wall cells and vestibular ganglion cells. These findings suggest that ET may play an important role in maintaining homeostasis of the vestibule. PMID- 8718531 TI - Observation of the otolithic membrane by low-vacuum scanning electron microscopy. AB - Untreated specimens (i.e. not fixed, dehydrated or embedded) of the otolithic membrane from the sacculus of guinea pigs were observed at the ultrastructural level by low-vacuum scanning electron microscopy (LVSEM). This technique revealed the presence of a 15- to 20-mu m-thick layer of an amorphous substance (the supraotolithic cupula zone) on the surface of the otoliths, which was not detectable by conventional methods. Elemental analysis of this substance revealed relatively high concentrations of oxygen, sodium, phosphorus, chlorine, potassium and calcium. This amorphous substance was thought to have a role in fixing the otoliths onto the sensory epithelium. In addition, the tips of the triangular portions of the otoliths were not sharp as shown by conventional SEM and were seen to be more rounded by LVSEM. PMID- 8718532 TI - Changes in the phosphorylation of neurofilament proteins in facial motoneurons following various types of nerve lesion. AB - This report defines the conditions for changes in the phosphorylation state of neurofilaments (NF) after facial nerve lesions. In adult control rats, few phosphorylated neurofilament (pNF) epitopes were stained (using SMI 31 antibodies) in a small subpopulation of facial motoneurons. After various types of mechanical lesion (nerve transection with and without attaching a metal clip to the proximal nerve trunk, nerve crush, combined trigeminal and facial nerve lesions) of the right facial nerve, pNF immunoreactivity transiently increased in most cell bodies of facial motoneurons on the operated side. This pNF 'reaction' started within 2 days after the operation and persisted up to 2 weeks but remained longer in those animals in which axonal regeneration had been prevented or delayed by attaching a metal clip to the proximal nerve stump. After botulinum toxin application into facial muscles (i.e. inhibition of synaptic transmission at motoric endplates) there was no increase in the amount of pNF after 4 and 10 days, but it appeared in facial nuclei 4 weeks after injection on both, the treated and the untreated side, i.e. during functional restitution. Selectively transecting the 'vibrissal part' of the trigeminal nerve induced no obvious changes in the pNF immunoreactivity in facial motoneurons, but a combined trigeminal-facial lesion did. Labeling nonphosphorylated epitopes (using SMI 32 antibodies) showed a slight decrease in immunoreactivity in the neuropil of the facial nucleus 15 days after nerve transection and fixing a metal clip on the proximal nerve stump. PMID- 8718533 TI - Identification of proliferating keratinocytes in middle ear cholesteatoma using the monoclonal antibody Ki-67. AB - Cholesteatoma epithelium is characterized by a keratinocyte dysregulation with aggressive growth subsequently destroying the middle ear mucosa. The monoclonal antibody Ki-67 recognizes a nuclear antigen expressed by cells in the G1, S, and G2/M phases being used to determine the growing cell fraction in tissue samples. Cryostat sections of skin and cholesteatoma biopsies were examined immunohistochemically for reactivity with Ki-67 using the alkaline phosphatase anti-alkaline phosphatase method. Nuclear staining was seen in a small number of keratinocytes located in the basal cell layer of normal auditory meatal skin. In contrast, numerous cells of the basal and suprabasal layers in cholesteatoma were found to react with Ki-67. A cytoplasmic staining was also observed in both skin and cholesteatoma. In cholesteatoma, the cytoplasmic staining was stronger. These results clearly show that cholesteatoma epithelium proliferates at a higher rate than normal epidermis, confirming the hyperproliferative behavior of cholesteatoma. PMID- 8718534 TI - Fibrin-glue-reinforced paper patch myringoplasty of large persistent tympanic membrane perforations in the guinea pig. AB - Investigators have attempted to simplify and improve myringoplasty. However, techniques have not been applied to large, chronic human tympanic membrane perforations. Fibrin glue has been shown to improve wound strength but has not been significantly utilized in myringoplasty. To evaluate the effectiveness of paper patch myringoplasty reinforced with fibrin glue, 15 guinea pigs underwent repeated myringectomy to form persistent (>50%) perforations. The right ears were repaired with a paper patch and fibrin glue (treatment group), and the left ears with a paper patch alone (control group). Of 16 surviving ears (9 right and 7 left), only 1, a left ear, failed to heal. In the control group, histology revealed a larger fibrous layer approaching greater total eardrum thickness than in the treatment group. Neither group was significantly different from a normal, nonoperated tympanic membrane in terms of overall thickness. Fibrin glue failed to demonstrate histologic evidence of increased cellular proliferation and possibly hindered growth. High spontaneous healing rates in guinea pigs undermine myringoplasty modeling. PMID- 8718535 TI - Hypoglossal-facial nerve anastomosis after acoustic neuroma resection: influence of the time anastomosis on recovery of facial movement. AB - Facial movement following hypoglossal-facial nerve anastomosis was investigated in 29 acoustic neuroma patients. The amount of facial movement was assessed using both the grading system of House and Brackmann and the revised grading scale of Yanagihara. The data were analyzed to determine the influence of the time elapsed between tumor resection and anastomosis upon recovery of facial movement. A slightly larger number of patients with delayed anastomosis (7-23 months) showed minimally poorer results than those with early anastomosis (within 3 months). However, these differences were not statistically significant. Moreover, there was no apparent relationship between the duration of facial nerve paralysis and the recovery of facial movement within either of these two groups. These results showed that hypoglossal-facial nerve anastomosis can be delayed up to 2 years following tumor resection with only minimal effect on the recovery of facial movement. PMID- 8718536 TI - Revision tympanoplasty: surgical findings and results in Riyadh. AB - A report on the surgical findings and long-term results of 68 revision tympanoplasties performed at King Abdul Aziz University Hospital over a period of 7 years is presented. Temporalis fascia was used in all operations, with an overall success rate of 85% of graft take up, with significant improvement of hearing in the majority of cases. The causes of immediate failure were found to be due to faulty surgical techniques, however most of the late reperforations were due to infections and in some to atrophy of the graft. It is concluded that better surgical technique and improvement of medical facilities in most rural areas are of utmost importance in Saudi Arabia. PMID- 8718537 TI - Cholesteatoma surgery with the canal-wall-down technique. AB - This is a retrospective study of a limited group of 30 patients, children and adults, diagnosed as having cholesteatomas. Analysis includes audiological data and surgical results. Two techniques were used in surgery. In half of the cases, an obliteration technique was used, and in the remainder an open radical cavity was created. The method used was determined by the surgeon and in some measure by the extent of the disease. Our conclusions are that the obliteration technique offers better hearing results than the open-cavity technique. The number of residuals over a 4- to 5-year period was acceptable and cavity problems were avoided. PMID- 8718538 TI - Spontaneous nystagmus in normal subjects. AB - We investigated the incidence of spontaneous nystagmus in 30 normal volunteers using electronystagmography (ENG). In all subjects, ENG was performed with the eyes open in the primary position, with the eyes closed, and with the eyes open in total darkness. Spontaneous nystagmus was detected in 5 (16.7%) of 30 subjects. In 15 subjects who underwent 2 ENG examinations, 4 subjects (26.7%) exhibited spontaneous nystagmus. Caloric testing was performed in 26 of 30 subjects. Spontaneous nystagmus was detected in 3 subjects who had a right-left difference in caloric response of over 40%, and in 2 of 23 subjects who had a difference of less than 40%. Our results showed that spontaneous nystagmus was detected in more subjects when the ENG examinations were repeated. Thus, a single ENG examination cannot exclude the possibility of spontaneous nystagmus. PMID- 8718539 TI - Surgical decompression in endocrine orbitopathy--a three-dimensional locating device ensures greater safety. AB - Numerous techniques have been described for surgical decompression in endocrine orbitopathy. Irrespective of the approach chosen, the procedure might involve hazards to important vessels and nerves near the base of the skull. When the optic nerve is compressed and relief of pressure at the orbital apex is necessary, the internal carotid artery and the cavernous sinus are at risk as well as the optic nerve itself. Although the use of modern optical instruments such as operating microscopes and endoscopes is now standard practice for this form of surgery, adequate topographical orientation is not achieved in every case. For this reason a computer-assisted locating device developed in this department has been employed by us in 7 cases of endocrine orbitopathy where surgical decompression was required. By using computer-tomographic or MR tomographic image data, the positions of structures at risk can be plotted to within 0.6 mm. Used in conjunction with an operating microscope and with endoscopic techniques, the system proved helpful in all cases. PMID- 8718541 TI - Fibrous dysplasia in paranasal cavities. AB - Presentation is made of 2 boys and a girl with fibrous dysplasia involving the frontal and ethmoidal sinuses and the maxilla. This tumor-like growth of the bone was not restricted to the sinus alone but expanded to the orbit, adjacent sinus, skull base and infratemporal or pterygoid fossa. In such cases, surgical therapy is the treatment of choice, with maximal preservation of healthy tissue and avoidance of major devastations that may result in undesired cosmetic defects. The etiology of the disease is still unknown. However, it should be emphasized that all 3 patients had suffered an injury to the adjacent bony structures at various time points preceding the disease manifestation. That is why we are inclined to believe that trauma might be a factor responsible for the onset of fibrous dysplasia. PMID- 8718540 TI - Cartilaginous incisions in septoplasty. AB - Although cartilaginous incisions are effective in straightening the deviation of the cartilaginous septum, the resultant morbidity is seldom addressed. We examined 1,164 patients who underwent septoplasty by the senior author. The patients were randomly subjected either to septal reconstruction by conventional cartilaginous incisions or to our modification of cartilaginous incisions. During the follow-up of 11 months to 7 years with an average of 2.5 years, saddle deformity of the external nose was observed in 10 of 567 patients (1.8%) who underwent septal reconstruction by conventional septal incisions, while no saddle nose deformity was observed in patients who underwent septal reconstruction by our modification of septal cartilaginous incision. Inattention to cartilaginous weakening from incisions may expose the patient to an unnecessary risk of complications. Nasal resistance on the narrow side of the nose was significantly decreased after septal reconstruction using the two techniques of cartilaginous incision. The results of this study suggest that our modification of cartilaginous incision may be an efficient, safe technique to decrease nasal resistance and to prevent saddle nose deformity of the external nose after septal surgery. PMID- 8718543 TI - [Effects of gonadotropin-releasing hormone and its analogue (buserelin) on aromatase in cultured human granulosa cells]. AB - We studied the direct effects of gonadotropin-releasing hormone (GnRH) and its analogue (buserelin) on aromatase activity and the aromatase cytochrome P-450 (P 450arom) concentration in cultured human granulosa cells which were obtained during oocyte retrieval for in vitro fertilization. Aromatase activity was assessed by radioassay with [1beta-3H]androstenedione as the substrate. The P 450arom concentration was determined by an enzyme-linked immunosorbent assay with specific antibodies to P-450arom. Buserelin stimulated aromatase activity and P 450arom at low concentrations (10(-13) - 10(-9)M), but it suppressed these parameters at high concentrations (10(-8) - 10(-7)M). The stimulatory effect increased with time during 12- to 48-h culture and disappeared after 72-h culture. Follicle-stimulating hormone (FSH) (100 ng/ml) stimulated aromatase activity and the P-450arom concentration but, FSH stimulation was suppressed by co-administration of low or high concentrations of buserelin. In contrast GnRH suppressed aromatase activity and P-450arom at all concentrations (10(-12) - 10( 6)M). GnRH also suppressed FSH stimulation of aromatase. Aromatase activity was correlated with the P-450arom concentration. These results suggest that in human preovulatory granulosa cells, buserelin and GnRH modulate aromatase activity by changing the P-450arom concentration. PMID- 8718542 TI - [Usefulness of estriol for the treatment of bone loss in postmenopausal women]. AB - To assess the effect of estriol (E3) on bone loss in Japanese postmenopausal women, twenty-five women within 10 years after menopause received 2 mg/day of E3 and 2 g/day of calcium lactate (Ca) daily (n = 17; Group-E3 + Ca) or 2 g/day of Ca daily (n = 8; Group-Ca) for one year. Bone mineral density (BMD) at the lumbar spine L2 - L4 was evaluated by dual energy X-ray absorptiometry (DXA). In Group Ca, BMD was significantly reduced (p < 0.05 vs. pretreatment) at the end of the study period. In contrast, in Group-E3 + Ca, the mean percent increase in BMD was 1.66%, which was significantly higher than that of Group-Ca (-3.08%; p < 0.001). As for the biochemical markers of bone metabolism, bone specific alkaline phosphatase (ALPIII) reflecting bone formation, and pyridinoline (Pyr) and deoxypyridinoline (D-Pyr) reflecting bone resorption, were significantly decreased (p < 0.001) in Group-E3 + Ca after six months of treatment. These data indicate that the acceleration of bone turnover usually observed after menopause was prevented by treatment with E3. As to the side effects of E3, only one of the 17 patients experienced a slight increase in vaginal discharge, but no other side effects were observed. Accordingly, reasonable compliance can be expected among postmenopausal women treated with E3 to prevent bone loss. PMID- 8718544 TI - [Significance of fetal arterial oxygen saturation (SpO2) monitoring]. AB - The significance of fetal arterial oxygen saturation (SpO2) monitoring and its relationship with FHR patterns were studied. Continuous fetal SpO2 was obtained by infrared pulse oximetry (Nellcor N-400) in 76 cases. After amniotomy, a reflectance fetal oximetry sensor (Nellcor FS-10) was placed onto the fetal cheek. The relationship between SpO2 and FHR pattern was then studied and the following results were obtained. 1. Mean recording time was 218 min. and satisfactory records were obtained during 75% of that time. 2. Whenever the FHR was reassuring, the range of SpO2 was stable within 40-80%. 3. Variations in SpO2 values were noted during decelerative patterns. When the FHR was not reassuring, the SpO2 value tended to be lower than 40%. A more acidotic tendency was found in cases where SpO2 was below 40% prior to delivery (umbilical arterial pH 7.135 +/- 0.100 vs 7.282 +/- 0.047, p < 0.005). 4. In cases of fetal arrhythmia (congenital complete heart block 3, supra ventricular tachycardia 1), SpO2 monitoring seemed to be an effective tool for fetal surveillance during labor. Fetal SpO2 measurement seemed to be an alternative to scalp blood sampling and is especially effective in cases with a nonreassuring or uninterpretable FHR pattern such as fetal arrhythmia. PMID- 8718545 TI - [An epidemiological study of malformations in infants delivered from diabetic mothers]. AB - The purpose of this study is to investigate the risks of delivery of malformed infants in diabetic mothers in comparison with those in non-diabetic mothers and to clarify risk factors for malformation due to maternal diabetes. The delivery records available at the Department of Obstetrics and Gynecology, Mie University School of Medicine between 1979 and 1992 showed 103 mothers with diabetes mellitus among 4,353 pregnancies. The incidence of malformation in infants born to diabetic mothers was 9.7%, and it was 2.5% overall. By logistic regression analysis of all mothers in this series, diabetes as well as maternal age, and fetal age were shown to be significant risk factors for malformation. A further logistic model for diabetic mothers revealed that a younger age of the mother at the time of labour, low fetal age, older age at onset of diabetes, previous history of pregnancies, and an Hemoglobin A1 (HbA1) level higher than 9% were independent risk factors, among which HbA1 (>9%) showed the highest odds ratio of 24. Careful management is therefore needed to reduce the risk of malformation in pregnancy of diabetic mothers. PMID- 8718546 TI - [Analysis of oxidative stress in rat fetus induced by clamping and reperfusion of uterine vessels]. AB - Effects of oxidative stress (OS) on the fetus were studied under ischemic induced by clamping the uterine vessels of pregnant rats. OS was evaluated by superoxide (O-.2) released from red blood cells. Fetal RBC-O-.2 of the ischemic rats (clamping for 10 min and 20 min) was significantly higher than that of the control group (p < 0.01). Maternal RBC-O-.2 tended to increase, but not significantly. When readings before and after clamping were compared, the activity of fetal RBC SOD had increased in the ischemic group (clamping time for 10 min), but significantly decreased in the 20 min clamping rats (p < 0.01). Maternal SOD activity tended to decrease along with clamping time. The amount of lipidperoxide was significantly greater in the fetal ischemic liver subjected to clamping for 20 minutes than in the tissue subjected to clamping for 10 minutes. The lipidperoxide level was significantly higher in the fetal ischemic brain exposed for 10 minutes than in the control. These results indicate that fetal RBC O-.2 increased along with ischemic time, and the maternal RBC-O-.2 had been influenced by the increase in fetal RBC-O-.2. Based on these observations, it was concluded that the OS due to ischemic injury had a more adverse effect on the fetal brain than on the liver. PMID- 8718547 TI - [Expression of glutathione S-transferase pi and numerical aberration of chromosome 11 in human ovarian cancer as a prognostic indicator]. AB - In 33 patients with primary ovarian cancer who had not received any chemotherapy before surgery, expression of glutathione S-transferase pi (GST-pi) was studied immunohistochemically in relation to the response to chemotherapy with CDDP, and furthermore it was examined whether numerical aberration of chromosome 11 was available as a prognostic indicator. The obtained results were as follows: 1. Of 33 ovarian cancer tissue samples, 19 (57.6%) showed positive staining and 14 (42.4%) negative staining for GST-pi. 2. The 5 year survival rate of the patients with GST-pi positive tumors was significantly lower than that of those with GST pi negative tumors (p < 0.01). 3. The frequency of detection of cells showing numerical aberration of chromosome 11 in tissues was significantly higher in ovarian cancers than in benign ovarian tumors and normal ovaries. 4. The 5 year survival rate of the patients with ovarian cancer in which the cells showing numerical aberration of chromosome 11 at 20% or more was significantly lower than that of those in which such cells were under 20% (p < 0.02). These results show that expression of GST-pi and numerical aberration of chromosome 11 in the tissues of human ovarian cancers are useful as prognostic indicators. PMID- 8718548 TI - [Cytotoxic action of monoclonal antibodies OC125 and SH-9 on ovarian cancer cell lines]. AB - A large number monoclonal antibodies (Mo-Abs) to neoplasms have been utilized as diagnostic tools, but they have not yet been fully characterized from the biological point of view. Antibody-dependent cellular cytotoxicity, complement dependent cytotoxicity (CDC) and antibody-dependent cell-mediated cytotoxicity (ADCC) were investigated in Mo-Abs OC125 and SH-9 (IgG1) to ovarian tumor cell lines SHIN-3, MN-1, KEN-3 and EC by MTT assay in vitro. OC125, but not SH-9 showed prominent CDC activity against SHIN-3 and MN-1 cell lines. CDC activity was induced against OC125 and SH-9, when a guinea pig complement was used. Furthermore, ADCC, activity was significant against SHIN-3, MN-1, OC125 and SH-9, but not against KEN-3 or EC. In summary, OC125 had both CDC and ADCC activity, but SH-9 did not. We concluded that the epitope on CA125 antigen varied in OC125 and SH-9, and each of epitope in Mo-Abs may have a different biofunction. PMID- 8718549 TI - [An immunohistological study on expression of glutathione S-transferase pi (form) in human endometrial carcinoma]. AB - Ninety-five specimens from patients with endometrial carcinoma (82 of endometrial type, 6 of adenoacanthoma, 4 of adenosquamous carcinoma, 3 of atypical endometrial hyperplasia) and 13 with ovarian endometrioid carcinoma were stained immunohistochemically with a rabbit polyclonal antibody prepared against the placental form of the enzyme glutathione S-transferase pi (GST-pi). Histological studies showed that the degree of staining decreased as the tumor lost its differentiation in endometrial carcinoma, but the degree of staining was independent of the differentiation in the case of ovarian endometrioid carcinoma. A comparison between the grade of staining of GST-pi in 82 cases of the endometrial type of endometrial carcinoma and 13 cases of ovarian endometrioid carcinoma revealed a stronger stain for the endometrial carcinoma than for ovarian endometrioid carcinoma (p < 0.01, Mann-Whitney's U test). Therefore, the GST-pi value for endometrial carcinoma was different from that for endometrioid carcinoma. In general, as compared with ovarian endometrioid carcinoma, endometrial carcinoma is considered to be resistant to chemotherapeutic agents. In conclusion, these results suggest that there is an apparent correlation between the GST-pi value and chemoresistance of the tumor. PMID- 8718550 TI - [Separation and characterization of estrogen independent endometrial cancer cell line originated from Ishikawa strain]. AB - An estrogen independent sub-clone of human endometrial carcinoma Ishikawa was established by culturing the mother cells in the absence of estrogen over a year. The daughter cells (designated as EIIL) possessed truncated estrogen receptors of an apparent size of 43Kd as well as normal 65Kd receptors and had lost sensitivity to estrogen or tamoxifen but remained sensitive to ICI 164,384. Analysis of the expression of the estrogen responsive element binding protein showed the presence of variant proteins which can recognize ERE. These observations indicate that long time estrogen deprivation may alter the expression of ER and this, in turn, alters cell response to estrogen as well as antiestrogens. PMID- 8718551 TI - [Apoptosis of prolactin induced structural luteolysis in rat]. PMID- 8718552 TI - [A case of giant uterine leiomyoma with chromosomal aberration]. PMID- 8718553 TI - [Laparoscopic procedures with abdominal wall-lift for ovarian cyst during pregnancy]. PMID- 8718554 TI - Effect of lupeol, a pentacyclic triterpene, on urinary enzymes in hyperoxaluric rats. AB - Investigations were undertaken to study the role of lupeol, a pentacyclic triterpene from Crataeva nurvala stem bark, in calcium oxalate experimental rat urolithiasis. A 2% solution of ammonium oxalate was administered by gastric intubation for inducing hyperoxaluric condition in adult male rats of Wistar strain. The duration of treatment was for 15 days. This resulted in increased urinary excretion of oxalate associated with reduction in citrate and glycosaminoglycans. The urinary marker enzymes which indicate renal tissue damage namely--lactate dehydrogenase, inorganic pyrophosphatase, alkaline phosphatase, gamma glutamyl transferase, beta-glucuronidase and N-acetyl beta-D glucosaminidase were found to be elevated. Lupeol administration (25 mg/kg body weight/day) reduced significantly the renal excretion of oxalate. It also reduced the extent of renal tubular damage as evidenced from the decreased levels of the above enzymes in urine. Such a reduction is likely to be beneficial in minimizing the deposition of stone-forming constituents in the kidney which provides antilithic effect. PMID- 8718555 TI - Induction of anti-tumor immunity by mouse tumor cells transfected with mouse interleukin-12 gene. AB - Interleukin-12 (IL-12) is a heterodimeric cytokine. In order to transduce both cDNAs for p35 and p40 of IL-12 in the tumor cells, a polycistronic retroviral vector was constructed by inserting the internal ribosome entry site gene of encephalomyocarditis virus between two cDNAs. On the other hand, two cDNAs were sequentially transfected in the tumor cells. Both polycistronic gene transfectants and double transfectants produced biologically active mouse IL-12. IL-12-expressing tumor cells were all rejected in syngeneic mice, and induced cytotoxic T lymphocyte activity. The capacity to induce anti-tumor memory may depend on the amount of IL-12 produced by the transfectants, because the relatively higher IL-12 producer tumor cell line induced the anti-tumor memory in the rejected mice, but the lower producer did not. PMID- 8718556 TI - Characterization of VP4 and VP7 of a murine rotavirus (YR-1) isolated in Japan. AB - Nucleotide and deduced amino acid sequences of the outer capsid proteins VP4 and VP7 of a murine rotavirus strain (YR-1) isolated in Japan were determined. Comparisons of the VP7 amino acid sequence of YR-1 with other murine rotavirus strains (EB, EW, EC, EL and EHP) (1) showed that YR-1 was highly homologous to EB, EW, EC, EL and EHP. Moreover, YR-1 was more closely related to strains representing G3 than to any other G type. Analysis of the VP4 amino acid sequence revealed that YR-1 was highly homologous to EB, EW, EC and EL [tentatively P17 (1)], and more closely related to EHP [tentatively P18 (1)] than to any other P type. Enzyme immunoassay with monoclonal antibodies against G types (KU-4 and BH49 for G1, S2-2G10 and BW36 for G2, YO-1E2 and BC5 for G3; and ST-2G7 and BE18 for G4) and against a P type (YO-1S3, KU-12H and YO-2C2 for P8) showed no reactivity. These results indicate that YR-1 is highly homologous to EB, EW, EC and EL. PMID- 8718557 TI - Titration of varicella-zoster virus DNA in throat swabs from varicella patients by combined use of PCR and microplate hybridization. AB - We devised a simple procedure for titration of varicella-zoster virus (VZV) DNA in throat swabs from varicella patients. DNA which was extracted from throat swabs, together with known copy numbers of a cloned VZV DNA fragment, were 10 fold serially diluted and used as template in PCR. The PCR products, after heat denaturation, again serially diluted in 1.5 M NaCl and adsorbed to microplate wells. Then, biotin-labeled DNA probes were hybridized with the immobilized DNA. The hybridization signal was produced by streptavidin-conjugated beta galactosidase and a fluorogenic enzyme substrate. By comparing the titration curves of a clinical specimen with those of the cloned fragment, of which detection limit was about 10 copies, we estimated the copy numbers of VZV DNA in the specimen. With this technique, we evaluated the degree of potential contagiousness of the patient along the course of infection: we found that varicella patients possessed highest quantity of VZV DNA in the throat on the first day of illness. PMID- 8718558 TI - L-2-Oxothiazolidine-4-carboxylate and N-acetylcysteine as precursors of intracellular glutathione in human peritoneal mesothelial cells. AB - L-2-Oxothiazolidine-4-carboxylate and N-acetylcysteine as substrates for intracellular glutathione in human peritoneal mesothelial cells were tested. Both substances at concentrations of 0.01 mM and higher augmented the level of glutathione in mesothelial cells. L-2-Oxothiazolidine-4-carboxylate had a milder but more stable effect than N-acetylcysteine. Cells with increased concentrations of the intracellular glutathione were more resistant to injury by free radicals. When used at higher concentrations (> 1 mM), both substances became cytostatic to mesothelial cells as evidenced by growth inhibition. PMID- 8718559 TI - Comments on the interpretation of tissue impedance measurements during hemodialysis. AB - Comparison of electrical impedance measured in vivo with the in vitro impedance of several types of tissue shows that the former is mainly determined by the presence of striated muscle in the region between the electrodes. This means that fluid shifts occurring during hemodialysis, in the blood or any tissue other than skeletal muscle, do not affect tissue impedance. This implies that measurement of tissue impedance as a means of monitoring the process of hemodialysis lacks a model for a reliable interpretation in terms of fluid balance. PMID- 8718560 TI - Acute suppression of parathyroid activity during hemofiltration. AB - Hemofiltration (HF) induces a significant reduction in parathormone (PTH). This effect is related not only to the convective removal of PTH molecules but also to the biological suppression of parathyroid glands by plasma-ionized calcium (iCa) increase. The acute inhibitory effect on parathyroid gland activity, ionized calcium mass balance, phosphate kinetics and intact PTH (PTHi) dialytic removal during post-dilution polyamide HF were studied in 31 chronic uremic patients. HF ensures good phosphate removal (from 2.54 +/- 1.19 to 1.27 +/- 0.35 mEq/l; p < 0.01), a positive iCa mass balance (8 +/- 4 mmol/session) with a iCa plasma increase and negligible convective PTHi removal (9 +/- 2 pg/ml). Study of the PTHi profile during HF characterized two different parathyroid responses: 26/31 patients showed a physiological parathyroid gland response to the iCa increase (from 1.17 +/- 0.09 to 1.42 +/- 0.07 mmol/l; p = 0.002) with a significant PTHi decrease (from 123 +/- 111 to 35 +/- 28 pg/ml; p = 0.01) and a maximal PTH inhibition of 88%. In 5 patients, with more severe hyperparathyroidism, in spite of a comparable increase in iCa (from 1.28 +/- 0.12 to 1.46 +/- 0.08 mmol/l; p = 0.02), this physiological calcium-PTHi feedback was lost, revealing an autonomization of the gland (maximal inhibition of 45%). In our experience, study of the PTHi profile during a single HF session may represent a clinical test for the functional exploration of parathyroid glands, suggesting future (medical or surgical) clinical strategy. PMID- 8718561 TI - Effects of slowly performed daytime hemodialysis (slow HD) on the pharmacokinetics of vancomycin in hemodynamically unstable patients with renal failure. AB - Effects of slowly performed daytime hemodialysis (slow HD) using a high-flux hemodialyzer on the pharmacokinetics of vancomycin were determined in 5 critically ill patients with renal failure. Following intravenous administration of 0.5 g of vancomycin, concentrations in the serum and dialysate were monitored. Pharmacokinetic parameters were calculated after fitting individual concentration time curves to a two-compartment model. The volume of distribution at steady state was 0.58 +/- 0.12 liters/kg. Total body clearance was 37.46 +/- 3.20 ml/min with an elimination phase half-life of 8.72 +/- 0.99 h. Slow HD clearance was 20.19 +/- 2.30 ml/min. During a 10-hour session of slow HD, the serum vancomycin concentration decreased from 44.2 +/- 3.8 to 10.0 +/- 5.0 mg/l and 30.10 +/- 5.34% of the dose was eliminated. Dialyzer clearance of this drug and urea was 18.71 +/- 1.40 and 28.77 +/- 1.77 ml/min, respectively. Slow HD may effectively eliminate vancomycin by a diffusive mechanism and this elimination should be taken into consideration for designing the dosage schedule during the treatment. PMID- 8718562 TI - Retention of cytokine-inducing substances inside high-flux dialyzers. AB - Reprocessing of dialyzers is often performed with nonsterile solutions possibly contaminated with bacterial-derived cytokine-inducing substances. We investigated the retention of cytokine-inducing substances inside the dialyzer during reprocessing in a closed loop in vitro hemodialysis system using a polyamide high flux membrane. After the first in vitro circulation of human whole blood, rinse of the blood compartment (BC) and reverse ultrafiltration (RUF) was performed with either cytokine-inducing substance-free saline or saline contaminated with filtrates from Pseudomonas cultures (6 ng/ml LAL-reactive material); subsequently, dialyzers were stored in 2% formaldehyde. Dialyzers were rinsed with approximately 15 liters pyrogen-free saline before the second circulation using blood from the same donor; the effluates were free of cytokine-inducing substances and formaldehyde. Before and after the blood circulations, peripheral blood mononuclear cells (PBMC) were separated and total production of IL-1 alpha and IL-1 beta was determined after overnight incubation. In noncirculated PBMC as well as in PBMC separated after whole blood circulation with pyrogen-free processed dialyzers, production of IL-1 beta was not detectable. After contaminated rinse of the BC, production of IL-1 beta could be observed (1,600 +/ 1,100 pg/ml, mean +/- SEM). When pyrogen-free RUF was performed after contaminated BC rinse, IL-1 beta production averaged 163 +/- 92 pg/ml when using reused dialyzers, but 1,820 +/- 880 pg/ml when using new dialyzers. After reuse with pyrogen-free BC-rinse and contaminated RUF no IL-1 beta synthesis was observed; however, when pyrogen-free BC-rinse and contaminated RUF was applied to new dialyzers, IL-1 beta synthesis averaged 1,620 +/- 1,200 pg/ml. We conclude that cytokine-inducing substances are retained inside the dialyzer, probably by adsorption to the membrane as well as to the protein layer covering the membrane and are still biologically active after sterilisation. Cytokine-inducing substances adsorbed to the protein layer can be partially removed by RUF. Finally, the protein layer on the membrane appears to reduce the convective transfer of cytokine-inducing substances from the dialysate into the blood compartment. PMID- 8718563 TI - Effect of cilazapril on hyperdipsia in hemodialyzed patients. AB - To investigate whether angiotensin-converting enzyme inhibitor (ACE-I) potentially alleviates hyperdipsia, the effect of cilazapril on dialysis associated excessive thirst was studied by evaluating various dipsogenic parameters in patients undergoing chronic hemodialysis (HD) who manifest an excessive interdialysis body weight gain of more than 5%, and show simultaneous severe-to-moderate hyperdipsia. An initial single dose of 1 mg of cilazapril given at the end of the HD session produced a marked improvement in the interdialysis thirst scores and a simultaneous reduction in plasma angiotensin II (AII) concentration due to the inhibition of ACE activity. The interdialysis body weight gain in the cilazapril treatment period was significantly smaller than that in the nontreatment period. None of the other parameters including blood pressure, plasma osmolarity, and serum Na and K concentration were different in the treatment vs. the nontreatment period. The present data help to explain the potential pharmacological action of AII in the physiology of thirst and suggest that cilazapril may effectively alleviate dialysis-associated hyperdipsia at least on some occasions. The mechanism by which ACE-I exerts an antidipsogenic action may, in part, be accounted for by the reduction in plasma concentration of AII, as a result of the ACE inhibition. PMID- 8718564 TI - Undetected excessive ultrafiltration during maintenance haemodialysis. AB - Undetected excessive ultrafiltration (UF) occurred in 2 patients on maintenance haemodialysis (HD) using a Gambro AK-10 monitor and Ergo 120 A-G blood lines. In spite of alarms present on the monitor, the dialysis staff was not alerted to the faulty condition. Through investigation of the HD 'set-up' at the time of the incidents it was found that a partial obstruction (kinking) had occurred in the extracorporeal blood circuit where the flexible tubing joined the venous chamber. This created a very high positive pressure within the dialyzer compartment and excessive UF, which the blood circuit pressure monitoring transducer was unable to detect. To prevent kinking of the blood lines, dialysis staff and patients should be instructed to ensure that blood lines lie in smooth curves. PMID- 8718565 TI - Dyslipidemia in renal disease patients. PMID- 8718566 TI - Diagnosis and classification of dyslipidemia in renal disease. AB - Renal disease is accompanied by specific alterations of the lipoprotein metabolism. While marked hyperlipidemia is a characteristic finding in the nephrotic syndrome, the dyslipoproteinemia of renal insufficiency is predominantly reflected in an abnormal apolipoprotein pattern but does not necessarily include elevated plasma lipid concentrations. The specific changes in nephrotic syndrome include increased formation primarily of cholesterol-rich and to a varying extent of triglyceride-rich ApoB-containing lipoproteins in the VLDL LDL density range with little or no change among the ApoA-containing lipoproteins in HDL. The dyslipoproteinemia of renal failure is, on the other hand, mainly characterized by a decreased catabolism of the triglyceride-rich ApoB-containing lipoproteins with increased concentrations of partially metabolized lipoproteins of intermediate and very low density and a decreased concentration of ApoA containing lipoproteins in HDL. In addition, increased levels of Lp(a) are found both in the nephrotic syndrome and in renal failure. Dialysis treatment appears to have only a modest influence on the renal dyslipoproteinemia. Due to its atherogenic character, the dyslipidemia of renal disease may be related to the accelerated development of cardiovascular disease in these patients. PMID- 8718567 TI - Do glomerular atherosclerosis and lipid-mediated tubulo-interstitial disease cause progressive renal failure in man? AB - The nephrotic syndrome presents the kidney with a new environment in which blood vessels, glomerular structures and tubules are exposed over substantial periods of time to lipoproteins. LDL has charge affinity with glomerular basement membrane glycosaminoglycans, so potentially increases or maintains albumin loss. This in turn stimulates LDL synthesis. HDL is small enough to be passed by the glomerular filter in substantial amounts and has been found to stimulate endothelin-1 production by human proximal tubular cells in culture. LDL also inhibits nitric oxide vasodilatory responses, an action which when added to that of endothelin-1 may result in decreased renal tissue oxygenation. Taken together, these aspects of the nephrotic syndrome broaden conventional definitions of atherosclerosis and offer a number of targets for therapy in progressive renal disease. PMID- 8718568 TI - Mechanisms of cardiovascular injury in renal disease. AB - Patients with renal disease have an increased cardiovascular mortality. Hyperlipidemia, a hallmark of renal disease, is recognized as a principal cause of atherosclerosis. However, it is difficult to prove a pathogenetic role of renal dyslipidemia per se in this increased cardiovascular risk since multiple risk factors are often present in patients with progressive renal insufficiency, e.g. hypertension, diabetes and hypercoagulability. However, evidence is accumulating demonstrating detrimental effects of hyperlipidemia during both initiation and progression of the atherosclerotic process. The present review discusses this evidence in patients with renal disease, and the possible implications for treatment. PMID- 8718569 TI - Treatment of dyslipidemia in chronic renal failure. AB - Dyslipidemia in end-stage renal disease is a common problem and may contribute to the high rates of morbidity and mortality in this population. Recent studies indicate that defective lipolysis is a major factor in the development of this disorder which is characterized by increased levels of very-low-density lipoprotein remnant particles, hypertriglyceridemia and occasionally hypercholesterolemia. There are no prospective long-term studies on the effect of lipid-lowering treatment on morbidity and mortality related to dyslipidemia. Therefore, at present pharmacologic treatment of hyperlipidemia should be undertaken in patients with severe hypertriglyceridemia (> 500 mg/dl) or hypercholesterolemia (LDL > 130 mg/dl) who are at high risk for coronary artery disease. This review discusses the pathogenesis of dyslipidemia, common clinical patterns of hyperlipidemia and various nonpharmacologic and pharmacologic treatment options. PMID- 8718570 TI - Treatment of multiorgan failure. PMID- 8718571 TI - Extracorporeal blood purification in the treatment of acute renal failure with multiorgan involvement. AB - It has been more than 35 years since the early use of dialytic techniques for the treatment of acute renal failure. Considering the survival of patients who would have clearly succumbed to renal failure, attempts were made to determine at what level of uremic toxicity it would be beneficial to offer dialysis on a prophylactic basis. Those early attempts at defining a preventive strategy seemed to provide an improved survival by limiting incidence and severity of hemorrhage and sepsis. Subsequently, measurable advances have been difficult to attain, and the prognosis of complicated acute renal failure remained dismally poor. More recently, continuous renal replacement techniques were developed to offer a more physiologic treatment to those patients who were the most critically ill and unstable. These treatments offered the potential for a better tolerated fluid removal and a more constant control of electrolyte and acid-base balance. Other potential advantages included the use of more biocompatible membranes and the convective mode of solute transfer with its inherently greater removal of larger molecular weight substances such as the vasodilatory and inflammatory cytokines. Another approach has been to use extracorporeal purification to lower the levels of endotoxin, a goal which can be accomplished by either standard plasma exchange or, more elegantly, the use of selective adsorption columns. Despite the promise of these methodologies, the lack of an acceptable grading system for the severity of critical illness has hampered investigators who have tried to prove the eventual advantage of one blood purification technique over another. PMID- 8718572 TI - Plasmapheresis in severe sepsis or septic shock. AB - During sepsis, there is release of various endotoxins from microorganisms which more or less activates cascade systems including release of cytokines such as tumor necrosis factor alpha and interleukin 6 and complement components. This causes impairment of vascular integrity and permeability which may progress into septic shock and a disseminated intravascular coagulation which progresses into multiorgan failure, including acute renal failure and subsequent death. Although most endotoxins and cytokines have a molecular size < 50 kD, there is little efficacy in removal of them by hemofiltration filters used for acute dialysis. The use of antibodies against different endotoxins has not been successful. The use of plasma exchange procedures (including blood exchange) to remove such toxins and cell debris, as free myoglobin and hemoglobin, has been successfully tried in smaller not controlled studies since 1984. Once when more than three organs are involved in a progressive manner, the risk of death is at least 80%. In contrast, these studies showed a survival rate of about 75% by addition of such therapeutic interventions to the conventional intensive care unit treatment. The substitution of the removed plasma products must be considered to include products important for the host defense and coagulation process and to avoid infections, bleeding, or increased coagulation. This type of removal is unselective and probably in the future will include addition of absorption techniques which may add further benefit to the outcome. PMID- 8718573 TI - Plasma exchange in patients with septic shock including acute renal failure. AB - In patients with septic shock and multiorgan failure including acute renal failure, the prognosis is poor. This study evaluated the effects of plasma exchange as adjunctive therapy to the conventional treatment in such severely ill patients and compared the results with the expected outcome according to data from other studies. A total of 25 patients (17 men and 8 women, mean age 47, range 15-74 years) were treated by a median of 3 (range 1-10) plasma exchanges, mainly by centrifugation technique. The main replacement fluid was liquid-stored plasma and albumin (in a few cases fresh frozen plasma). About 80% of the patients also received low doses of steroids (hydrocortisone 200-400 mg/day and heparin 1,000-25,000 U/day). Twenty of the patients survived (80%) which was significantly better than was expected by conventional treatment (< 20% survival, p < 0.001). The relative chance to survive was four times greater than expected. Five patients died. The reason of death was cerebral hemorrhagia, brain abscess, myocardial sudden death, relapsing sepsis from multiple hepatic abscesses, and a psoas abscess which was not drained. The patients who survived regained almost total renal function and could leave the hospital with only a few sequelae. The plasma exchange may be a good adjunct to conventional therapy to increase the chance of survival when other treatment is insufficient. The mechanisms by which plasma exchange acts may be by removal of endotoxins derived from microorganisms or tissue necrosis and removal of excessive amounts of cytokines together with a modification of various cascade systems enabled by the products in the plasma from the healthy donors, used for replacement. PMID- 8718574 TI - Nutritional aspects in patients with acute renal failure/multiorgan failure. AB - Nutrition support contributes to reducing morbidity and mortality in patients with the multiple-organ dysfunction syndrome (MODS). In cases when acute renal failure is a component of MODS, the goals of nutritional management should be to control general nutrient deficiencies and correct specific metabolic alterations caused by the loss of renal function or acute uraemia without worsening uraemic toxicity or disturbing electrolyte balance. In most cases energy requirements, which should ideally be estimated by indirect calorimetric measurements, amount to 30 kcal/kg/day and only seldom exceed 35 kcal/kg/day. Depending on the degree of protein catabolism, 1.0-1.5 g/kg/day of protein should be given. A balanced amino acid solution containing both essential and nonessential amino acids should be administered. Uraemia impairs protein synthesis and nitrogen administration should, therefore, not be started until the uraemic state is adequately controlled by dialysis or haemofiltration. PMID- 8718575 TI - Synthetic vaccines for infectious and autoimmune diseases. PMID- 8718577 TI - Hybrid hepatitis B virus core antigen as a vaccine carrier moiety. II. Expression in avirulent Salmonella spp. for mucosal immunization. AB - Hepatitis B virus (HBV) core antigen (HBcAg) is a highly immunogenic subviral particle. We and others have defined insertion sites for heterologous epitopes and successfully used hybrid particles to generate B and T cell immunity (reviewed in: Schodel et al. 1994a, 1995). Here we shall review recent progress in constructing avirulent Salmonella spp. expressing hybrid HBcAg particles carrying different epitopes. Hybrid HBcAg particles carrying virus neutralizing epitopes of the hepatitis B virus pre-S region or repeat epitopes of plasmodial circumsporozoite antigens were previously described (Schodel et al. 1992, 1994b). Salmonella spp. can be attenuated by defined genetic means so that they become avirulent, yet preserve invasiveness after oral uptake. Hybrid HBcAg-pre-S particles were expressed in Salmonella typhimurium and S. typhi vaccine strains. A single oral immunization of mice with such live recombinant S. typhimurium strains elicited a high titered serum anti-pre-S1 IgG response. Similarly, circumsporozoite repeat epitopes of three different malaria parasites were expressed as HBcAg-CS hybrids in recombinant S. spp. and were found to be highly immunogenic after oral immunization. To analyze mucosal immune responses, BALB/c mice were immunized with recombinant phoPc S. typhimurium expressing HBcAg by various mucosal routes (Hopkins et al., 1995). All routes of immunization resulted in high titered serum and local antibodies against HBcAg and S. typhimurium LPS. However, nasal immunization was most efficient in generating pulmonary IgA and rectal immunization in eliciting rectal IgA, suggesting some compartmentalization of the mucosal immune response. PMID- 8718576 TI - Host range restricted, non-replicating vaccinia virus vectors as vaccine candidates. AB - Three model systems were used to demonstrate the immunogenicity of highly attenuated and replication-defective recombinant MVA. (1) Intramuscular inoculation of MVA-IN-Fha/np induced humoral and cell-mediated immune responses in mice and protectively immunized them against a lethal respiratory challenge with influenza virus. Intranasal vaccination was also protective, although higher doses were needed. (2) In rhesus macaques, an immunization scheme involving intramuscular injections of MVA-SIVenv/gag/pol greatly reduced the severity of disease caused by an SIV challenge. (3) In a murine cancer model, immunization with MVA-beta gal prevented the establishment of tumor metastases and even prolonged life in animals with established tumors. These results, together with previous data on the safety of MVA in humans, suggest the potential usefulness of recombinant MVA for prophylactic vaccination and therapeutic treatment of infectious diseases and cancer. PMID- 8718578 TI - Synthetic recombinant vaccine induces anti-influenza long-term immunity and cross strain protection. PMID- 8718579 TI - Alphavirus-based expression systems. PMID- 8718580 TI - Alphavirus hybrid virion vaccines. PMID- 8718581 TI - DNA vaccines for bacteria and viruses. PMID- 8718582 TI - New vaccines against bacterial toxins. PMID- 8718583 TI - Parameters for the rational design of genetic toxoid vaccines. PMID- 8718584 TI - Protective immunity induced by Bacillus anthracis toxin mutant strains. PMID- 8718585 TI - Bacterial outer membrane protein vaccines. The meningococcal example. PMID- 8718586 TI - Changing paradigms for an HIV vaccine. PMID- 8718587 TI - Complexed HIV envelope as a target for an AIDS vaccine. PMID- 8718588 TI - HIV-peplotion vaccine. A novel approach to vaccination against AIDS by transepithelial transport of viral peptides and antigens to Langerhans cells for induction of cytolytic T cells by HLA class I and CD1 molecules for long term protection. PMID- 8718589 TI - The role of adjuvants and delivery systems in modulation of immune response to vaccines. PMID- 8718590 TI - Unique immunomodulating properties of dimethyl dioctadecyl ammonium bromide (DDA) in experimental viral vaccines. PMID- 8718591 TI - Challenges in the development of combination vaccines. PMID- 8718592 TI - Polysaccharide conjugate vaccines for the prevention of gram-positive bacterial infections. PMID- 8718593 TI - Production of influenza virus in cell cultures for vaccine preparation. AB - Influenza virus strains of different types for use as an inactivated vaccine have been successfully grown in different cell lines. Increasing titres were obtained with BHK-21/BRS, VERO and MDCK cells. Cultures in stationary flasks, in spinner cultures or in large bioreactor systems were tested and the optimal conditions were studied. MDCK cells grown in serum-free medium before and during the virus production phase were found to yield high titres in the presence of trypsin. Satisfactory results were obtained with egg-adapted strains of human and equine origin as well as with strains just isolated from human patients without any further passages in eggs or cell culture. PMID- 8718594 TI - Analysis of Bordetella pertussis suspensions by ELISA and flow cytometry. PMID- 8718595 TI - Clinical trials of Shigella vaccines in Israel. PMID- 8718596 TI - Hypothesis: how licensed vaccines confer protective immunity. AB - By examining experience with evaluation of licensed vaccines we theorize that a critical level of serum IgG confers protection against infectious diseases by killing or inactivating the inoculum. We found that efficacy is reliably predicted by measurement of serum antibodies elicited by vaccines, that serum IgG antibodies alone account for the protection conferred by passive immunization, that vaccine-induced "herd" immunity is best explained by inactivation of the inoculum on epithelial surfaces by serum antibodies and that serum antibodies induced by active immunization will neither treat disease symptoms nor eliminate the pathogen. If valid, this theory should facilitate research because knowledge of the pathogenesis of the disease symptoms may not be essential for vaccine development. PMID- 8718597 TI - Therapeutic vaccines. A pandoric prospect. PMID- 8718598 TI - Antimicrobial resistance: a global perspective. PMID- 8718599 TI - Resistance to antibacterial agents. PMID- 8718600 TI - Bacterial resistance to carbapenems. AB - The carbapenems have the broadest spectra of all beta-lactams but resistance still occurs, caused by target modification, impermeability or beta-lactamase production. Target modification or replacement is important in methicillin resistant staphylococci, E. faecium and some pneumococci. These organisms present the greatest current threat to carbapenem efficacy. Impermeability to carbapenems arises in P. aeruginosa mutants, where it is contingent on loss of D2 outer membrane protein, a minor porin. This resistance functions only if the Pseudomonas retains its chromosomal group 1 beta-lactamase, and so reflects the interplay of impermeability and hydrolysis rather than impermeability alone; nevertheless porin loss is the critical change that engenders resistance. Resistance based on impermeability coupled to a group 1 beta-lactamase has also been described in E. cloacae, but demands loss of a major porin and is much rarer and less stable than in P. aeruginosa. Although group 1 beta-lactamases contribute to resistance in these organisms, they have only feeble carbapenemase activity. Chromosomal beta-lactamases with potent carbapenemase activity occur in most or all X. maltophilia, A. hydrophila and F. odoratum isolates. These enzymes, which cause carbapenem resistance when expressed copiously, are all zinc dependent. Zinc carbapenemases also are a concern in B. fragilis, where they are encoded by the chromosomal DNA of c. 3% of isolates, though expressed by only 1%. Carbapenemases are extremely rare outside these species. Nevertheless, a plasmidic zinc carbapenemase was reported from one P. aeruginosa isolate and from several S. marcescens. Further carbapenemases, some not zinc-dependent, are known from a tiny numbers of Serratia, Enterobacter, and Acinetobacter isolates. Despite these various modes of resistance, carbapenems have retained their efficacy far better than have expanded-spectrum cephalosporins. Whether this advantage will be retained indefinitely is uncertain. If resistance does become more prevalent it may be possible to derivatize the carbapenems so as to extend their activity. There is already interest in the design of carbapenems that bind beta-lactam-resistant PBPs and, to an extent, in the development of carbapenemase inhibitors. PMID- 8718601 TI - Bacterial resistance to fluoroquinolones: mechanisms and patterns. PMID- 8718602 TI - A comparison of active site binding of 4-quinolones and novel flavone gyrase inhibitors to DNA gyrase. AB - The activity of 4-quinolone antibacterials at the enzyme target level is based on the well known and reported observations that 4-quinolone antibacterials target the Gyr A subunit of the DNA gyrase holoenzyme, inhibiting supercoiling while facilitating the "cleavable complex". Such inhibition can be observed by running the in vitro DNA gyrase supercoiling inhibition assay or the "cleavable complex" DNA gyrase assay. Although potency of the gyrase inhibitor is dependent on many factors including permeability and pharmacokinetics, the inherent potency of a gyrase inhibitor lies in its activity against the target enzyme. We have examined the binding activity of novel flavones [Bioorganic & Med. Chem. Letters 3:225 230, 1993] to Escherichia coli DNA gyrase and have found differences in binding consistent with inhibition of DNA gyrase supercoiling and ability to facilitate the cleavable complex, but of different rank order. [3H]norfloxacin was used in vitro competition studies with test compounds, pBR322 and E. coli DNA gyrase. Binding affinity results indicate the rank order of greatest to weakest binding (ability to compete with [3H]norfloxacin) of test compounds: Levofloxacin = ciprofloxacin > ofloxacin > norfloxacin > flavone compounds (including ellagic acid, quercetin, and compounds 5a through 5n [Bioorganic & Med. Chem. Letters 3:225-230, 1993]). Such differences in binding ability of the 4-quinolones and flavones to the ternary complex of DNA.DNA gyrase.drug, as compared to the catalytic inhibition and "cleavable complex" data, suggests a more complex binding of flavones than the previously hypothesized models for 4-quinolone binding. PMID- 8718603 TI - A low-copy number plasmid mediating beta-lactamase production by Xanthomonas maltophilia. AB - To delineate the mechanisms contributing to the high level of antimicrobial resistance often demonstrated by Xanthomonas maltophilia, plasmid DNA was isolated from 5 clinical isolates and analyzed. Purified plasmid DNA from a single isolate contained a 6.5 kb plasmid (pXM222) and a 5.6 kb plasmid, (pTHB). Transformation of pTHB into E. coli HB101 resulted in the expression of resistance to all penicillins tested and cefazolin. PMID- 8718604 TI - Glycopeptide resistance in gram-positive pathogens. PMID- 8718605 TI - Vancomycin-dependent Enterococci: a clinical and laboratory assessment. PMID- 8718606 TI - Streptococcus pyogenes: resistant, tolerant, neither or both? PMID- 8718607 TI - The antimicrobial susceptibility of Streptococcus pyogenes isolates from the Philadelphia area. PMID- 8718608 TI - Antibiotic-resistance in Streptococcus pneumoniae. PMID- 8718609 TI - Emerging trends in antimicrobial resistance: a laboratory perspective. PMID- 8718610 TI - Molecular epidemiologic analysis of antibiotic resistant microorganisms. PMID- 8718611 TI - Patterns of resistance in organisms causing gynecologic infections. PMID- 8718612 TI - Practical management of patients infected with resistant organisms: an infection control perspective. PMID- 8718613 TI - Resistance to anti-human immunodeficiency virus therapeutic agents. PMID- 8718615 TI - An improved method for in vitro susceptibility testing of Trichomonas vaginalis. AB - A protocol is presented for in vitro susceptibility testing of Trichomonas vaginalis. A 100 ml culture of the microorganisms is prepared for inoculation into antibiotic dilutions and controls by centrifugation, washing with 10 mM HEPES (pH 6.2) plus 1.5 x 10(-1) M NaCl, a second centrifugation and a resuspension in the HEPES-saline buffer. Inclusion of the gelling agent carrageenan in the culture medium permits an ease of harvesting the trichomonads and a reproducible initial cell density of 1-4 x 10(4) cells per ml. Following inoculation, tubes with antibiotic dilutions and controls are incubated anaerobically at 35 degrees C for 48 h, which corresponds to late exponential phase. Inclusion of a negative control helps determine Minimum Lethal Concentration (MLC) values. PMID- 8718614 TI - Molecular approaches to the spreading problem of drug resistant malaria. PMID- 8718616 TI - Antifungal drugs and resistance. PMID- 8718618 TI - Control of flower development and phyllotaxy by meristem identity genes in antirrhinum. AB - The flower meristem identity genes floricaula (flo) and squamosa (squa) promote a change in phyllotaxy from spiral to whorled in Antirrhinum. To determine how this might be achieved, we have performed a combination of morphological, genetic, and expression analyses. Comparison of the phenotypes and RNA expression patterns of single and double mutants with the wild type showed that flo and squa act together to promote flower development but that flo is epistatic to squa with respect to early effects on phyllotaxy. We propose that a common process underlies the phyllotaxy of wildtype, flo, and squa meristem development but that the relative timing of primordium initiation or growth is altered. This process depends on two separable events: setting aside zones for potential primordium initiation and partitioning these zones into discrete primordia. Failure of the second event can lead to the formation of continuous double spirals, which are occasionally seen in flo mutants. PMID- 8718617 TI - Tissue-specific accumulation of MURB, a protein encoded by MuDR, the autonomous regulator of the Mutator transposable element family. AB - The Mutator (Mu) system of transposable elements is highly mutagenic and can maintain high levels of activity through multiple generations due to frequent transpositions of both its autonomous and nonautonomous components. This family also shows pronounced developmental regulation. Most notable is the very low frequency of germinal reversions, despite the high levels of somatic transpositions and excisions, and the high frequency of germinally transmitted duplication events. Here, we report the production of antibodies raised against MURB, one of two proteins encoded by MuDR, the autonomous regulator of the Mu family. Immunolocalizations performed using anti-MURB antibodies reveal that this protein is present in specific tissues during male inflorescence development. Throughout much of development, MURB is detected at the highest levels in cell lineages that may find themselves in the germ line, but no MURB is detected in microspore mother cells. These cells are the direct precursors to pollen. Based on these observations as well as previous data, we discuss the relationship between the expression of MURB and developmental regulation of Mu activity. PMID- 8718619 TI - Immunocytological localization of an epitope-tagged plasma membrane proton pump (H(+)-ATPase) in phloem companion cells. AB - In higher plants, the plasma membrane proton pump (H(+)-ATPase) is encoded by a surprisingly large multigene family whose members are expressed in different tissues. Using an 18-amino acid epitope tag derived from the animal oncogene c Myc, we have performed immunocytolocalization measurements of the protein expressed by one member of this family, AHA3 (Arabidopsis H(+)-ATPase isoform 3). Immunofluorescence studies with tissue sections of transgenic plants have revealed that c-Myc-tagged AHA3 is restricted to the plasma membrane of phloem companion cells, whereas other AHA isoproteins are more widely distributed in the plasma membrane of other cell types. Electron microscopy with immunogold-labeled tissue sections suggests that there is a high concentration of proton pumps in the plasma membrane of companion cells but a much lower concentration in the plasma membrane of sieve elements. Due to plasmodesmata connecting the plasma membrane of these two adjacent cell types, it is likely that the proton motive force generated by the proton pump in companion cells can serve to power the uptake of sugar by proton-coupled symporters in either the companion cell or sieve element cell. The abundance of the proton pump in the plasma membrane of companion cells supports an apoplastic model for phloem loading in which the metabolic energy that drives sugar uptake is consumed by AHA3 at the companion cell plasma membrane. These experiments with a genetically altered integral plasma membrane protein demonstrate the utility of using a short c-Myc sequence as an epitope tag in Arabidopsis. Furthermore, our results demonstrate that, using genes encoding individual members of a gene family, it is possible to label plasma membrane proteins immunologically in specific, differentiated cell types of higher plants. PMID- 8718620 TI - Tobacco mosaic virus movement protein-mediated protein transport between trichome cells. AB - Tobacco mosaic virus movement protein (TMV MP) is required to mediate viral spread between plant cells via plasmodesmata. Plasmodesmata are cytoplasmic bridges that connect individual plant cells and ordinarily limit molecular diffusion to small molecules and metabolites with a molecular mass up to 1 kD. Here, we characterize functional properties of Nicotiana clevelandii trichome plasmodesmata and analyze their interaction with TMV MP. Trichomes constitute a linear cellular system and provide a predictable pathway of movement. Their plasmodesmata are functionally distinct from plasmodesmata in other plant cel types; they allow cell-to-cell diffusion of dextrans with a molecular mass up to 7 kD, and TMV MP does not increase this size exclusion limit for dextrans. In contrast, the 30-kD TMV MP itself moves between trichome cells and specifically mediates the translocation of a 90-kD beta-glucuronidase (GUS) reporter protein as a GUS::TMV MP fusion. Neither GUS by itself nor GUS in the presence of TMV MP moves between cells. These data imply that a plasmodesmal transport signal resides within TMV MP and is essential for movement. This signal confers selectivity to the translocated protein and cannot function in trans to support movement of other molecules. PMID- 8718621 TI - Tobacco mosaic virus movement protein associates with the cytoskeleton in tobacco cells. AB - Tobacco mosaic virus movement protein P30 complexes with genomic viral RNA for transport through plasmodesmata, the plant intercellular connections. Although most research with P30 focuses on its targeting to and gating of plasmodesmata, the mechanisms of P30 intracellular movement to plasmodesmata have not been defined. To examine P30 intracellular localization, we used tobacco protoplasts, which lack plasmodesmata, for transfection with plasmids carrying P30 coding sequences under a constitutive promoter and for infection with tobacco mosaic virus particles. In both systems, P30 appears as filaments that colocalize primarily with microtubules. To a lesser extent, P30 filaments colocalize with actin filaments, and in vitro experiments suggested that P30 can bind directly to actin and tubulin. This association of P30 with cytoskeletal elements may play a critical role in intracellular transport of the P30-viral RNA complex through the cytoplasm to and possibly through plasmodesmata. PMID- 8718622 TI - Molecular characterization of the CER1 gene of arabidopsis involved in epicuticular wax biosynthesis and pollen fertility. AB - The aerial parts of plants are coated with an epicuticular wax layer, which is important as a first line of defense against external influences. In Arabidopsis, the ECERIFERUM (CER) genes effect different steps of the wax biosynthesis pathway. In this article, we describe the isolation of the CER1 gene, which encodes a novel protein involved in the conversion of long chain aldehydes to alkanes, a key step in was biosynthesis. CER1 was cloned after gene tagging with the heterologous maize transposable element system Enhancer-Inhibitor, also known as Suppressor-mutator. cer1 mutants display glossy green stems and fruits and are conditionally male sterile. The similarity of the CER1 protein with a group of integral membrane enzymes, which process highly hydrophobic molecules, points to a function of the CER1 protein as a decarbonylase. PMID- 8718623 TI - End-to-end annealing of plant microtubules by the p86 subunit of eukaryotic initiation factor-(iso)4F. AB - The p86 subunit of eukaryotic initiation factor-(iso)4F from wheat germ exhibits saturable and substoichiometric binding to maize microtubules, induces microtubule bundling in vitro, and is colocalized or closely associated with cortical microtubule bundles in maize root cells, indicating its function as a microtubule-associated protein (MAP). The effects of p86 on the growth of short, taxol-stabilized maize microtubules were investigated. Pure microtubules underwent a gradual length redistribution, an increase in mean length, and a decrease in number concentration consistent with an end-to-end annealing mechanism of microtubule growth. Saturating p86 enhanced the microtubule length distribution and produced significantly longer and fewer microtubules than the control, indicating a facilitation of annealing by p86. Confirmation of endwise annealing rather than of dynamic instability as the mechanism for microtubule growth was made using mammalian MAP2, which also promoted the redistribution of length, increase in mean length, and decrease in number concentration of taxol stabilized maize microtubules. Enhancement of microtubule growth occurred concomitant with bundling by p86, indicating that an alignment of microtubules in bundles facilitated endwise annealing kinetics. The results demonstrate that nonfacile plant microtubules can spontaneously elongate by endwise annealing and that MAPs enhance the rate of annealing. The p86 subunit of eukaryotic initiation factor-(iso)4F may be an important regulator of microtubule dynamics in plant cells. PMID- 8718624 TI - Genetic dissection of carotenoid synthesis in arabidopsis defines plastoquinone as an essential component of phytoene desaturation. AB - Carotenoids are C40 tetraterpenoids synthesized by nuclear-encoded multienzyme complexes located in the plastids of higher plants. To understand further the components and mechanisms involved in carotenoid synthesis, we screened Arabidopsis for mutations that disrupt this pathway and cause accumulation of biosynthetic intermediates. Here, we report the identification and characterization of two nonallelic albino mutations, pds1 and pds2 (for phytoene desaturation), that are disrupted in phytoene desaturation and as a result accumulate phytoene, the first C40 compound of the pathway. Surprisingly, neither mutation maps to the locus encoding the phytoene desaturase enzyme, indicating that the products of at least three loci are required for phytoene desaturation in higher plants. Because phytoene desaturase catalyzes an oxidation reaction, it has been suggested that components of an electron transport chain may be involved in this reaction. Analysis of pds1 and pds2 shows that both mutants are plastoquinone and tocopherol deficient, in addition to their inability to desaturate phytoene. Separate steps of the plastoquinone/tocopherol biosynthetic pathway are affected by these two mutations. The pds1 mutation affects the enzyme 4-hydroxyphenylpyruvate dioxygenase because it can be rescued by growth on the product but not the substrate of this enzyme, homogentisic acid and 4 hydroxyphenylpyruvate, respectively. The pds2 mutation most likely affects the prenyl/phytyl transferase enzyme of this pathway. Because tocopherol-deficient mutants in the green alga Scenedesmus obliquus can synthesize carotenoids, our findings demonstrate conclusively that plastoquinone is an essential component in carotenoid synthesis. We propose a model for carotenoid synthesis in photosynthetic tissue whereby plastoquinone acts as an intermediate electron carrier between carotenoid desaturases and the photosynthetic electron transport chain. PMID- 8718625 TI - Meiotic recombination break points resolve at high rates at the 5' end of a maize coding sequence. AB - Sequence analysis of recombination break points has defined a 377-bp recombination hot spot within the anthocyanin 1 (a1) gene. One-fifth of all recombination events that occurred within the 140-kb a1-shrunken 2 interval resolved within this 377-bp hot spot. In yeast, meiotic double-strand breaks in chromosomal DNA are thought to initiate recombination and are generally located 5' of coding regions, near transcription promoter sequences. Because the a1 recombination hot spot is located within the 5' transcribed region of the a1 gene, the sites at which recombination events initiate and resolve appear to be different, but both appear to be regulated in relation to transcribed sequences. Although transposon insertions are known to suppress recombination and alter the ratio of crossovers to apparent gene conversions, the Mutator 1 transposon insertion in the a1-mum2 allele does not alter the sites at which recombination events resolve. PMID- 8718626 TI - Targeting and topology in the membrane of plant 3-hydroxy-3-methylglutaryl coenzyme A reductase. AB - The enzyme 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) catalyzes the synthesis of mevalonate. This is the first committed step of isoprenoid biosynthesis. A common feature of all known plant HMGR isoforms is the presence of two highly conserved hydrophobic sequences in the N-terminal quarter of the protein. Using an in vitro system, we showed that the two hydrophobic sequences of Arabidopsis HMGR1S function as internal signal sequences. Specific recognition of these sequences by the signal recognition particle mediates the targeting of the protein to microsomes derived from the endoplasmic reticulum. Arabidopsis HMGR is inserted into the microsomal membrane, and the two hydrophobic sequences become membrane-spanning segments. The N-terminal end and the C-terminal catalytic domain of Arabidopsis HMGR are positioned on the cytosolic side of the membrane, whereas only a short hydrophilic sequence is exposed to the lumen. Our results suggest that the plant HMGR isoforms known to date are primarily targeted to the endoplasmic reticulum and have the same topology in the membrane. This reinforces the hypothesis that mevalonate is synthesized only in the cytosol. The possibility that plant HMGRs might be located in different regions of the endomembrane system is discussed. PMID- 8718627 TI - Plant inositol monophosphatase is a lithium-sensitive enzyme encoded by a multigene family. AB - myo-Inositol monophosphatase (IMP) is a soluble, Li(+)-sensitive protein that catalyzes the removal of a phosphate from myo-inositol phosphate substrates. IMP is required for de novo inositol synthesis from glucose 6-phosphate and for breakdown of inositol trisphosphate, a second messenger generated by the phosphatidylinositol signaling pathway. We cloned the IMP gene from tomato (LeIMP) and show that the plant enzyme is encoded by a small gene family. Three different LeIMP cDNAs encode distinct but highly conserved IMP enzymes that are catalytically active in vitro. Similar to the single IMP from animals, the activities of all three LeIMPs are inhibited by low concentrations of LiCl. LeIMP mRNA levels are developmentally regulated in seedlings and fruit and in response to light. Immunoblot analysis detected three proteins of distinct molecular masses (30, 29, and 28 kD) in tomato; these correspond to the predicted molecular masses of the LeIMPs encoded by the genes. Immunoreactive proteins in the same size range are also present in several other plants. Immunolocalization studies indicated that many cell types within seedlings accumulate LeIMP proteins. In particular, cells associated with the vasculature express high levels of LeIMP protein; this may indicate a coordinate regulation between phloem transport and synthesis of inositol. The presence of three distinct enzymes in tomato most likely reflects the complexity of inositol utilization in higher plants. PMID- 8718628 TI - Phytochrome A and phytochrome B mediate the hypocotyl-specific downregulation of TUB1 by light in arabidopsis. AB - Arabidopsis contains six alpha-tubulin and nine beta-tubulin genes that are expressed in a tissue-specific and developmentally regulated manner. We analyzed the effects of light on tubulin mRNA abundance in Arabidopsis seedlings using RNA gel blot hybridizations and gene-specific probes. Transcript levels of all 15 tubulin genes were decreased by continuous white light, although to different degrees. Detailed analysis was performed with the beta-tubulin TUB1 gene. The transcript level of TUB1 was high in etiolated seedlings and decreased to approximately 20% of the dark mRNA level after 2 to 6 hr of white light treatment. We showed that this downregulation requires high-irradiance light treatment and that multiple photoreceptors are involved. In particular, using phytochrome mutants and narrow wave band light, we demonstrated that both the phytochrome A (phyA)-mediated far-red light high-irradiance response and the phytochrome B (phyB)-mediated red light high-irradiance response are involved in the downregulation of TUB1 expression by white light. Histochemical analysis of transgenic plants expressing a TUB1-beta-glucuronidase chimeric transgene indicated that the downregulation observed only in hypocotyls and not in roots is controlled transcriptionally. PMID- 8718629 TI - Interactions between distinct types of DNA binding proteins enhance binding to ocs element promoter sequences. AB - Octopine synthase (ocs) elements are a group of promoter elements that have been exploited by plant pathogens to express genes in plants. ocs elements are components of the promoters of certain plant glutathione S-transferase genes and may function as oxidative stress response elements. Genes for ocs element binding factors (OBFs), which belong to a specific class of highly conserved, plant basic domain-leucine zipper transcription factors, have been isolated and include the Arabidopsis OBF4 and OBF5 genes. To characterize proteins that modulate the activity of the OBF proteins, we screened an Arabidopsis cDNA library with the labeled OBF4 protein and isolated OBP1 (for OBF binding protein). OBP1 contains a 51-amino acid domain that is highly conserved with two plant DNA binding proteins, which we refer to as the MOA domain. OBP1 is also a DNA binding protein and binds to the cauliflower mosaic virus 35S promoter at a site distinct from the ocs element in the 35S promoter. OBP1 specifically increased the binding of the OBF proteins to ocs element sequences, raising the possibility that interactions between these proteins are important for the activity of the 35S promoter. PMID- 8718631 TI - [Variants in the arrangement of the tooth roots in the alveolar arches of the human jaws based on computed tomographic data]. AB - Anatomo-topographic relationship of the teeth roots were studied by means of computer tomography (CT) on 44 passportized skulls with mandibles of people who died in the age of 20 to 52 years. Conseguently, sections of mandibles and maxillae were made out of these preparations and peculiarities of the teeth roots locations in alveolar arches were studied by craniometric method. Comparison of CT results and measurements of sections showed that CT is an informative method of assessment of the teeth roots anatomo-topographic relationship that permits to attain general true qualititative indexes, allowing to evaluate objectively the available variants of the alveolar arch structure and the teeth roots location without disturbance of the anatomic material integrity. PMID- 8718630 TI - [The characteristics of the structural development of the thymus and spleen in the progeny of white rats exposed to pyrogenal action during pregnancy]. AB - Thymus and spleen structural changes of progeny of experimental animals after single exposure to 100-150 mg/l kg of bacterial pyrogenal lipopolysaccharide on 11th, 14 and 18th days of pregnancy were studied in the experiment performed on 60 female outbred albino rats and 205 rats--pups aged 20- and 45 days. Relative thymus and spleen masses were defined, their morphofunctional state was assessed morphometrically. To reveal the pecularities of the immune system reaction to foreign antigens in progeny, one part of control and experimental rats were subjected to immunization with ram erythrocytes (T-dependent antigen) and another one--with pyrogenal (T-independent antigen). Prenatal pyrogenal effect retarded thymus and spleen formation. Normalization of their structure and compensation of the effect were incomplete and occurred by 45th day of postnatal development. Most expressed changes were observed in the progeny of the rats treated with pyrogenal on 14th day of pregnancy: sharp decrease of the population density of lymphoid cells occurred in the thymus while lymphoid nodules disappeared in the spleen in response to immune challenge. PMID- 8718632 TI - [The morphofunctional characteristics of the deciduous membrane in pregnancy edema]. AB - An analysis of DNA content in decidua cells and decidua cell number in compact decidual layer was performed in 19 women with physiological pregnancy and labour, in 31 women with oedema of pregnancy, in 29 women with light and in 11 women with severe pre-eclampsia. Significant difference in DNA content in the group with oedema of pregnancy and in the groups with other gestosis forms was demonstrated. Thus, a conclusion is made that oedema may have an independent genesis. PMID- 8718633 TI - [The cytoarchitectonic characteristics of the speech center of the brain in gifted people in the plan to study individual variability of human brain structure]. AB - Cytoarchitectonics of cerebral cortical fields 44 and 45 was studied in gifted people in comparison with the group of people without creative professional talents. Frontal paraffin sections 20 mkm thick were examined. Both histological and modern quantitative methods were used with the aid of "Videoplan" computer. Peculiarities in the cytoarchitectonics of cortical fields 44 and 45 and, in particular, in expression of the nerve cell horizontal and vertical orientation and in the characteristics of the certain cortical fields delimination were demonstrated. Increase of pyramidal neuron glial index and of peculiarities of neurons regrouping in cytoarchitectonic layers III and IV was noted. PMID- 8718634 TI - [Accessory groups of nonapeptidergic cells in the diencephalon of intact and hypophysectomized rats]. AB - Accessory groups (AG) of oxytocin (OT)--and vasopressin (VP)--ergic cells (circular, perifornical, ventro- and dorsolateral groups and periventricularly localized nonapeptidergic cells) were studied morphometrically in intact and hypophysectomized rats. Additionally, the AG of medulla stria thalamus was demonstrated and investigated. The total number of nonapeptidergic cells in the AG is about 600 cells. The AG consist of OT-ergic cells predominantly (58%) except the perifornical group (68%) VP-ergic cells) and ventrolateral group (equal propotion). 7 days after hypophysectomy the size of perikaryon, nucleus and nucleolus of the cells in AG decreased, though, in contrast to the supraoptic and paraventricular hypothalamic centres, no degeneration of cell occured. That is why it is possible to suggest, that axons of the AG cells do not project to the posterior pituitary. The absence of hypertrophy of the AG cells in compensatory function of degenerating cells in the supraoptic and paraventricular nuclei indicates a functional diversity of these nonapeptidergic cells. PMID- 8718635 TI - [The ultrastructure of the myelinated fibers of the rat endbrain in hypokinesia]. AB - Myelin fibres structure in phylogenetically different regions of telencephalon- in hippocampus, cingular cortex and sensomotor cortex of cerebrum of rats, in hypokinesia lasting for 40 and 90 days was studied electron-microscopically. Changes in ultrastructure of myelin fibres in these formations were found which exposed unequally in different regions and depended on the experiment duration. In 40 days hypokinezia changes were noted only in hippocampus and cingular cortex, while in neocortex they were prominent only in 90-days hypokinisia. Moreover, in neocortex changes were different in large and fine fibres, being more prominent in the former. PMID- 8718636 TI - [The delta sleep-inducing peptide as a modulator of synaptic ultrastructure]. AB - Results of morphometric study of ultrastructure of axosomatic and axospinous synapses of rat sensomotor neocortex after DSIP administration and in hypoxia showed that the peptide administration resulted in multidirectional changes in these type contacts in intact animals. This, apparently, is connected with their different functional specialization. Preliminary DSIP administration prevented changes, appearing in hypoxia in most of the parameters studied both in axosomatic and axospinous synapses. PMID- 8718637 TI - [Projections of the tegmental ventral field, substantia nigra formations and amygdaloid nuclei to different segments of the caudate nucleus and nucleus accumbens in the dog]. AB - Spatial organization of progections of amygdala, tegmentum ventral field and of substantia nigra formations on different regions of caudal nuclei and of the nucleus accumbens in dog was studied by means of retrograde axonal transport of horseradish peroxidase. Distribution of progectional fibres of neurons of the structures studied in striatum allowed to reveal its dorsal and ventral zones. Neurons of compact and dorsal parts of substantia nigra and scanty amygdala neurons project on the dorsal zone. Fibers from neurons of all the formations of substantia nigra, ventral field and of amygdala basal nucleus project on ventral field. Intersection of projections of limbic and motor subcortical formations in corpus striatum is discussed in connection with interaction of functionally different systems in it. Data obtained in dog and cat were compared. PMID- 8718638 TI - [The development of transplants of human embryonic neocortex in the anterior chamber of the rat eye]. AB - Dorsolateral regions of embryonic human neocortex at 9-11 weeks of development were transplanted into the eye anterior chamber of rat. Significant changes, leading to the complete destruction of the original cytoarchitectonies occured during the first three days. Intensive neuroblast degeneration in cortical and interstitial zones as well as decrease of mitotically deviding cell numder were observed 3-6 hours after the transplantation. Neuroepithelial cells form rosettes in ventricular zone by the end of the first day. Rosettes turned into the centres of proliferation, mitotic activity restored to the initial level. Cells migrated out of the rosettes in radial directions. By the end of the third day almost all the cells in cortical and interstitial zones degenerated, transplant was almost completely represented by the ventricular zone cells. Number of mitotic figures exceeded the initial level almost three times, transplant was enlarged, its blood supply was absent. Since the 4-th day mitotic activity decreased, transplant underwent lymphoid infiltration and its complete destruction occured by the end of the second week. PMID- 8718640 TI - [The ultrastructural changes in the sympatheic ganglion in experimental hypercholesterolemia]. AB - In conditions of experimental atherisclerosis mature Chinchilla rabbits acquire dystrophic changes of different severeness in neurons, synapses, nervous fibres, perineurium and blood vessel endothelium of the superior cervical sympathetic ga (SCSG) during one month. It is suggested that B-lipoproteins penetrate easily th of SCSG capillaries (of sinusoidal type) and the diffuse neurotissue barrier (pe which leads to structural changes in neuron bodies, axons and synapses. SCSG neurons++ with CNS is disturbed as well as axonal catecholamine transport and neurotrophic of innervated magistral vessels (carotid arteries, aortal arch etc.). PMID- 8718639 TI - [Regional and cellular distribution of mitochondrial high-affinity aldehyde dehydrogenase in the rat brain (an immunocytochemical study)]. AB - Immunocytochemical study of regional and cellular distribution of mitochondrial high-affinity aldehyde dehydrogenase (MA AlDH) in the rat CNS was performed by streptavidin-biotin-peroxidase method. Specific immunoreactivity of diverse intensity was found in most structures of brain and spinal cord regions mainly in neuronal and gliocyte cytoplasm and to a smaller extent, in their processes, terminations (neuropile conducting tracts, white matter) and blood vessels: immune reaction intensity of perikarya of different type neurons exceeds those of neuropile 1.8-3.3 times. MA AlDH positive neurons were observed in all the cerebral regions, with the most intensive specific staining in inferior olives and hippocamp pyramidal layer (0.500-0.520 optical density units) and the minimal in nucleus rubrum and substantia nigra (0.260-0.290 units). In both cases MA AlDH immunoreactive neurons made only part (from 40 to 88) of morphologically identified (thianine-positive) neurons. Comparative analysis of histochemical preparations and the cytophotometric results revealed significant diversities in topographic distribution of MA AlDG and general AlDG activity in the cerebrum. The data obtained assist in elucidation the spatial organization of alcohol and aldehyde metabolism in CNS and in comprehension the reasons of different sensitivity of brain structures to alcohol. PMID- 8718641 TI - [The topographic anatomy of the nerves supplying the human heart]. AB - Investigations, performed in organ complexes of neck and mediastinum in 50 corpses of adults, demonstrated, that all cervical cardiac nerves and cardiac branches of vagal nerves are united into a single plexus, branches of which form the nerves, approaching the heart. Some of them are formed under the aortal arch and behind the brachiocephalic trunk. They are located on the ascending aorta and on the pulmonary trunk, approaching the coronary sulcus of the heart, participating in the innervation of all its regions. Other preorgan cardiac nerves are formed under the pulmonary arteries, approach the atria, ramifying in their walls and also participating in the forming the subepicardial plexuses, innervating diaphragmal surfaces of the heart ventricles. PMID- 8718642 TI - [The innervation of the conducting and working myocytes in the sinoatrial area of the canine heart (a quantitative study)-]. AB - Quantitative electron microscopic study of the innervation of different types of specialized sinus node conducting myocytes and of working myocytes in the perinodal myocardium of the right atrium was carried out in intact dogs. Differences were found in diameters of unmyelinated nervous fibers and their efferent terminals and distances between them and sarcolemmas of myocytes of the types studied. It was demonstrated that qualitatively different synaptic vesicles in the efferent terminals near the nodal and working myocytes are encountered with different frequency. Similar parameters of innervation of the nodal and working myocytes in sinauricular region of the heart were compared in intact dog and rat. PMID- 8718643 TI - [The structural reorganization of the myocardium in white rats under exposure to high temperature]. AB - The investigation was performed in 64 male albino rats, 18 of which stayed in comfortable conditions (control), while 48 were kept everyday for 4 hrs in open sun ground at temperature 30-40 degrees C during 3 months. Vascular-structural reorganizations in the myocardium of experimental animals developed during 1-15 days. Capillary-tissue disturbances increase with characteristic morphological manifestations of the blood circulation disorders prevailing in the left and right ventricles, sharp increase of vessel and capillary wall permeability, loosening of the organ stroma, dystrophic and degenerative processes in cardiomyocytes, presence of diapedese haemorrhages and decrease of glycogen content. From day 16 up to day 30 capillary-tissue disorders and manifestations of the structural-anatomic reorganizations subside although intramural blood circulation changes persist as well as single degenerative changes in cardiomyocytes and oedematous changes of the connective tissue framework. At later stages, up to day 90 the formed structural anatomical reorganizations stabolize, the signs of blood circulation disturbances disappear to significant extent, but complete normalization of microscopic myocardial structure does not occur. PMID- 8718644 TI - [The ultrastructural characteristics of the reactive changes in the cardiomyocytes of adult rats when cultured in vivo]. AB - 60 implantes of the left atrium myocardium were studied on the 1st, 3rd and 6th days of in vivo culture according to F. M. Lazarenko method (outbred laboratory male rats of 200-250g. body weight were used both as donors and recipients). Reactive changes of the cultured cardiomyocytes were assessed by methods of light and electron microscopy. Polymorphic nature of the cardiomyocyte reactive changes with the preservation of its tissue pecific determination was established. Myocardium implant did not form myosymplastic elements, cellular characteristics of organization remained in its implanted pieces. Data concerning degenerating and surviving cardiomyocytes are presented as well as those on cellular forms, identification of which is difficult on the ultrastructural level. PMID- 8718645 TI - [Individual and age-related variability of the arteries to the atrioventricular node in the human heart]. AB - Investigation was performed on 501 preparations of the hearts of people of various age without the pathology of cardiovascular system. Complex of morphological methods, including morphometry was used in the work. Individual variability of the atrioventricular node artery was established to be characterized by diverse diameter and length (within limits, determined for every age), direction, branching characteristics, zones of distribution, and different participation in blood supply of the atrioventricular region of conducting system of the heart. PMID- 8718646 TI - [The morphological characteristics and deformability and strength properties of the common carotid artery in fetuses and newborns]. AB - General carotid arteries were studied in 51 corpses of foetuses and newborns with gestational ages ranging from 28 to 40 weeks. The entire material was divided into 3 groups: 1) 28-32 weeks, 2) 33-36 weeks, 3) 37-40 weeks. The length of the cervical region as well as external diameter of the artery, thickness of the wall and its certain layers, number of elastic membrane rows and connective tissue fibres architectonics were studied. General strength, its limits, maximum relative deformation and arterial wall Jung module were investigated at longitudinal extension in tension machine. All the arterial morphological and strength characteristics increase in proportion with gestation age but maximum accretion takes place by the end of the last month of intrauterine development. In all age groups vascular wall extensibility is almost equal. Arterial wall has an elongation reserve due to the wavy course of elastic fibres in tunica media. PMID- 8718647 TI - [Irreversible ischemic damages in the skeletal muscle of the severed extremity depending on the duration of the ischemia and the temperature]. AB - On the model of allotransplantation of m. lumbricalis, previously subjected to acute total ischemia at temperature 37-38 degrees it was shown that morphological integrity and characteristic immunohistochemical properties of the muscle remain only during two hours of the thermic ischemia. A week later surviving muscular fibres (MF) and newly forming muscular tubules (MT) were found in the transplants of the muscle. Processing with antibodies of the fast twitch type myosin filaments revealed both fast- and slow-twitch types of fibres among MF, while B- and C-typed MF were identified at defining the activity of succinat dehydrogenase (SDG). High SDG activity, corresponding to C-type fibres was observed in MT. Staining with antibodies revealed fast-twitch type myosin. After 3-hour ischemia of the muscles, few surviving MF were found, while after the 4-6-hours one only MT appeared to be present in the transplants. Thus, refering to our data, obtained in the previous works and to the results of the research it is ascertained that there is a regularity, allowing to predict critical terms of development of irreversal ischemic disturbances in the muscle of severed extremity, depending on the ischemia duration and temperature. PMID- 8718649 TI - [A histochemical method for studying the activity of monoamine oxidase A and B in the brain]. PMID- 8718648 TI - [The adaptation of human skeletal muscle fibers during aerobic training]. AB - Effect of training on dimensions of muscle fibres of fast twitch and slow-twitch types were studied in three groups of subjects: local training of m. triceps surae (volunteers) in laboratory conditions, training of amateur joggers and of elite racers--skiers. Significant negative correlations between initial level and training-induced changes of muscle fibre cross sectional areas were demonstrated. The correlations found indicate that muscle fibre dimensions of people using similar endurance training programme tend to some certain level which is likely to be the optimal for this type of training. Dimensions of muscle fibre may significantly change as the result of traiining, probably, only in the case, when their initial values differ from the optimal one. PMID- 8718650 TI - [A method and device for taking organ and tissue samples for histological study]. PMID- 8718651 TI - [The use of magnetic resonance tomography in studying the central nervous system]. PMID- 8718652 TI - [A new method of fixing histological preparations using dimethylphosphite]. PMID- 8718653 TI - [Annali Italiani di medicina interna: present and future]. PMID- 8718654 TI - [Indication for transcranial Doppler studies in transient ischemic attack]. AB - The diagnostic procedures for the assessment of transient ischemic attacks (TIA) include both brain imaging (computed tomography, nuclear magnetic resonance) and cerebrovascular investigation by means of ultrasound studies and angiography. Transcranial Doppler sonography (TCD) has not yet become a widely used diagnostic tool, although it allows noninvasive investigation of the intracranial cerebral circulation. The aim of this study was to assess the value of TCD investigation in patients who suffered from TIA. Eighty-one consecutive patients admitted to our General Medicine Department with a diagnosis of TIA underwent cranial computed tomography, ultrasound investigation of the extracranial cerebral arteries, and TCD. RESULTS: 39% of the TCD findings were not significant; TCD findings were highly significant in 18% of the patients with TIA, mainly for intracranial arterial stenoses and symptomatic subclavian steal; the other abnormal TCD findings were nonspecific, but in all these cases TCD yielded information on the efficiency of intracranial collateralization and the mechanisms regulating cerebral hemodynamics. These results suggest that TCD is a useful tool for the assessment of TIA. PMID- 8718655 TI - [Quantitative ultrasonography in the evaluation of postmenopausal osteoporosis. Comparison with dual energy x-ray absorptiometry]. AB - In order to compare the capacities of quantitative ultrasonography and dual energy X-ray absorptiometry to measure bone mineral density (BMD) in postmenopausal women, 45 postmenopausal patients were studied. They were divided into two groups on the basis of the results of absorptiometry: Group A (20 patients) had BMD values within normal limits, and Group B (25 patients) had BMD values lower than 1 standard deviation (SD) of the average for an age and sex matched population. Ultrasonography of the non-dominant heel was carried out on both groups to determine broadband ultrasound attenuation (BUA), speed of sound (SOS) and bone velocity (BV). A negative correlation between age and BMD, BV, SOS, and BUA was observed in all subjects. A positive correlation between BMD and BV and between BMD and SOS was found. In Group A, age correlated negatively with BMD, BUA, SOS and BV, and BMD correlated positively with BV. In Group B, age correlated negatively with BMD, and BMD correlated positively with BV and SOS. BV values were significantly elevated in Group B. The study evidenced a relationship between ultrasonography and absorptiometry in determining bone density in postmenopausal women. As ultrasonography and absorptiometry provided comparable information on bone density, we point out that ultrasonography can be used as a simple and sensitive indicator for postmenopausal osteoporosis. PMID- 8718656 TI - [Malnutrition in the acute care hospital: a very common problem]. AB - The adequacy of caloric intake of geriatric patients in medical and surgical wards in the acute care hospital was assessed in a prospective, observational study. Fifty-one surgical and 80 medical nonterminal patients aged over 70 years underwent a multidimensional assessment on admission and a nutritional reassessment on discharge. The average daily caloric intake was also measured. Patients were divided into two groups according to whether the ratio of the actual to the needed caloric intake was inferior to 40% or not, and their differences, with regard to baseline values, were assessed. Patients whose caloric intake was inferior to 40% of the needed were older than the remaining ones (79.4 +/- 6.6 vs 76.6 +/- 4.9 years, p < 0.05), had higher prevalence of preadmission functional dependency (21.6% vs 15%, p < 0.001), lower body mass index (22.9 +/- 5.4 vs 24.8 +/- 3.9, p < 0.004) and higher comorbidity (coexisting diseases: 3.6 +/- 1.9 vs 2.8 +/- 1.6, p < 0.02) on admission. In conclusion, a simple assessment on admission allows targeting geriatric patients at risk for in-hospital starvation. No difference exists between surgical and medical wards in the quality of nutritional support. PMID- 8718657 TI - [Early diagnosis of rapidly progressive glomerulonephritis]. AB - The term rapidly progressive glomerulonephritis (RPGN) designates a group of glomerular diseases with different pathogenetic and clinical features, rapidly leading to renal or patient death in about 90% of the untreated cases. Histopathologically, it is characterized by glomerular crescents in at least 50 75% of the glomeruli (necrotizing crescentic glomerulonephritis), and very often, glomerular necrosis. The situation is, however, potentially reversible if adequately treated, and a favourable outcome depends largely on early diagnosis and treatment. Early diagnosis can be achieved if due importance is given to even seemingly unspecific manifestations such as "flu like syndrome" associated with "glomerular" hematuria. These manifestations are detectable before the down-hill course of renal functional derangement becomes evident and should lead the physician to consider RPGN among the diagnostic possibilities. Final diagnosis rests on serological tests and kidney biopsy. The battery of diagnostic serological tests (anti-GBM, anti-DNA antibodies, cryoglobulins, etc.) has recently been enriched by the assay of anti-neutrophil cytoplasmic antibodies (ANCA). These antibodies are detectable in over 90% of cases of Wegener's granulomatosis and primary necrotizing crescentic glomerulonephritis with or without lung involvement. ANCA-associated glomerulonephritis is the commonest form of RPGN, and the new serological assay provides an important tool for its early recognition. Renal biopsy is necessary to evaluate the severity of the nephritic process and modulate treatment accordingly. Timely diagnosis is one of the most important factors contributing to successful treatment outcome over both the short and the long term. PMID- 8718658 TI - [Brain natriuretic peptide]. AB - Brain natriuretic peptide (BNP) is a cardiac hormone with a spectrum of activities quite similar to those of atrial natriuretic peptide (ANP), including diuretic, natriuretic, hypotensive and smooth muscle relaxant activities. These effects are due to the stimulation of guanylate cyclase-linked natriuretic peptide receptors, leading to an increase in cyclic GMP concentration in target cells. BNP has a lower affinity than ANP for C (clearance) receptors, and is less susceptible to degradation by neutral endopeptidase-24.11, resulting in a longer half-life. In the kidney, BNP increases the glomerular filtration rate and inhibits sodium reabsorption in the distal tubule. It also inhibits the release of renin and aldosterone. Unlike ANP, produced by the atria, BNP is mainly synthesized and released into circulation by the left ventricle and is therefore influenced by stimuli involving this cardiac chamber, such as an increase in arterial pressure, left ventricular hypertrophy and dilation. Plasma BNP levels are very low in healthy subjects, and respond modestly, although significantly to physiological stimuli such as changes in posture or sodium intake. In contrast, plasma BNP concentrations increase in disease states such as cirrhosis with ascites, hypertension, chronic renal failure, acute myocardial infarction and congestive heart failure. In the latter condition, plasma BNP concentration is a reliable prognostic index. Evidence obtained by administering BNP to healthy subjects and hypertensive patients suggests that BNP, at physiological and pathophysiological plasma concentrations, markedly influences cardiovascular homeostasis, mainly due to its effects on sodium excretion and the renin aldosterone axis. PMID- 8718659 TI - [Changes of lung volumes and respiratory muscle strength in patients with growth hormone deficiency]. AB - The relationship between growth hormone deficiency (GHd) and ventilatory function is not well understood. We studied 7 patients with GHd since childhood who had been adequately treated with replacement therapy until cartilage fusion. Together with 7 well-matched (age and body-type) healthy control subjects, they underwent spirometry including determination of residual volume, and lung diffusing capacity. Also recorded were maximal respiratory muscle pressure during inspiration (PImax) and expiration (PEmax). Patients with GHd showed a significant reduction in total lung capacity and vital capacity while residual volume and lung diffusing capacity remained unchanged. All patients had a significant reduction of both PImax and PEmax. Previously treated adult subjects with GHd present a persistent decrease in lung mobilizing volumes associated with reduced respiratory muscle strength. These alterations may have implications in the management of GHd in adult patients. PMID- 8718660 TI - [Intestinal bleeding from an ulcerated diverticulum of the small intestine. A clinical case]. AB - A case of occult bleeding of the upper intestinal tract in a patient with secondary polycythaemia and aortic valve stenosis is described. Routine tests failed to disclose the origin of the bleeding and the patient's condition continued to deteriorate. On the 23rd day after admission, laparotomy was performed, and an ulcerated diverticulum diagnosed. Resection of the diverticulum was done immediately. The patient had an uneventful post-operative recovery and was dismissed from the hospital 15 days later. Our experience underscores the necessity for early laparotomy in cases of gastrointestinal bleeding of obscure origin. PMID- 8718661 TI - [Cardiac involvement in familial amytrophic chorea with acantocytosis: description of two new clinical cases]. AB - Familial amyotrophic chorea and acanthocytosis, also known as the Levine Critchley syndrome, is a rare inherited disease characterized primarily by central nervous system involvement with progressive demyelinization and autosomic or dominant transmission. Clinical symptoms include orofaciolingual dyskinesia and involuntary choreiform movements associated with skeletal muscle atrophy due to axonal demyelinization and erythrocyte acanthocytosis. A few patients have some cardiac abnormality, including an electrocardiographic pattern of left ventricular hypertrophy, left atrial wave abnormalities, non-specific ST-T wave changes, and a pseudonecrosis pattern with abnormal Q waves in the inferior leads. Two-dimensional echocardiography has disclosed concentric ventricular hypertrophy and the typical findings of congestive cardiomyopathy. We report the case of two brothers, 40 and 58 years old, who had asymmetric left ventricular hypertrophy (more marked in the younger brother), left ventricular mass index increase unrelated to a hypertensive state or the percent of circulating acanthocytes. Functional systolic parameters were normal. The younger brother had dilation of the aortic root and marked enlargement of the non-coronary Valsalva sinus, and both patients manifested mitral leaflet redundancy without evident prolapse. Our observations suggest the hypothesis that connective tissue and/or vessel muscle-elastic fiber pathology is associated with the well-known neurological disorders typical of the Levine-Critchley syndrome. It is thus advisable that these patients undergo thorough cardiovascular evaluation. PMID- 8718662 TI - [Meningeal carcinomatosis from pulmonary adenocarcinoma: two clinical cases with review of the literature]. AB - The case of a 68-year-old woman with lung adenocarcinoma complicated by apparently isolated widespread meningeal metastasis is described. Meningeal carcinomatosis is a rare complication of solid neoplasms. In fact, it occurs in less than 2% of lung carcinoma which is the most frequent cause of this syndrome. Although computed tomography and nuclear magnetic resonance may aid the diagnosis of this condition, only the cytological demonstration of neoplastic cells in the cerebrospinal fluid directly establishes it. In this patient, the onset of symptoms due to meningeal carcinomatosis complicated the course of a diagnosed lung adenocarcinoma with the acute onset of a confusional state and extended meningeal dissemination without clinical or instrumental findings of metastasis in other areas of the body. Acute confusional state is seldom the initial symptom of meningeal carcinomatosis. Nevertheless, in the neoplastic patient, this possibility must be taken into account after the most common causes of acute confusion have been excluded. Even when modern diagnostic imaging techniques are utilized, diagnosis must be confirmed by cytological examination of the cerebrospinal fluid. PMID- 8718663 TI - Mechanism of endothelial cell shape change and cytoskeletal remodeling in response to fluid shear stress. AB - Endothelium exposed to fluid shear stress (FSS) undergoes cell shape change, alignment and microfilament network remodeling in the direction of flow by an unknown mechanism. In this study we explore the role of tyrosine kinase (TK) activity, intracellular calcium ([Ca2+]i), mechanosensitive channels and cytoskeleton in the mechanism of cell shape change and actin stress fiber induction in bovine aortic endothelium (BAE). We report that FSS induces beta actin mRNA in a time- and magnitude-dependent fashion. Treatment with quin2-AM to chelate intracellular calcium release and herbimycin A to inhibit TK activity abolished BAE shape change and actin stress fiber induction by FSS, while inhibition of protein kinase C with chelerythrine had no effect. Altering intermediate filament structure with acrylamide did not affect alignment or F actin induction by FSS. Examining the role of the BAE cytoskeleton revealed a critical role for microtubules (MT). MT disruption with nocodazole blocked both FSS-induced morphological change and actin stress fiber induction. In contrast, MT hyperpolymerization with taxol attenuated the cell shape change but did not prevent actin stress fiber induction under flow. Mechanosensitive channels were found not to be involved in the FSS-induced shape change. Blocking the shear activated current (IK.S) with barium and the stretch-activated cation channels (ISA) with gadolinium had no effect on the shear-induced changes in morphology and cytoskeleton. In summary, FSS has a profound effect on endothelial shape and F-actin network by a mechanism which depends on TK activity, intracellular calcium, and an intact microtubule network, but is independent of protein kinase C, intermediate filaments and shear- and stretch-activated mechanosensitive channels. PMID- 8718664 TI - Axonemal tubulin polyglycylation probed with two monoclonal antibodies: widespread evolutionary distribution, appearance during spermatozoan maturation and possible function in motility. AB - Two monoclonal antibodies, AXO 49 and TAP 952, probed with carboxy-terminal peptides from Paramecium axonemal tubulin and with polyglycylated synthetic peptides, are found to recognize differently tubulin polyglycylation, the most recently identified posttranslational modification discovered in Paramecium axonemal tubulin. With these antibodies, we show that tubulin polyglycylation is widely distributed in organisms ranging from ciliated protozoa to mammals; it arose early in the course of evolution, but seems to be absent in primitive protozoa such as the Euglenozoa. Tubulin polyglycylation is the last posttranslational modification which takes place in the course of Drosophila spermatogenesis and its occurrence corresponds to the end of spermatozoan maturation. An involvement of polyglycylated tubulin in axoneme motility is suggested since AXO 49 and TAP 952 specifically inhibit the reactivated motility of sea urchin spermatozoa. PMID- 8718665 TI - Cycloheximide-induced activation of mouse eggs: effects on cdc2/cyclin B and MAP kinase activities. AB - Fertilization of metaphase II-arrested mouse eggs results in resumption of meiosis and a decrease in both cdc2/cyclin B kinase and MAP kinase activities; the decrease in cdc2/cyclin B kinase activity precedes the decrease in MAP kinase activity. Cycloheximide treatment of metaphase II-arrested mouse eggs also results in resumption of meiosis but bypasses the fertilization-induced Ca2+ transient. However, it is not known if cycloheximide treatment results in the same temporal changes in cdc2/cyclin B kinase and MAP kinase activities that are intimately associated with resumption of meiosis. We report that cycloheximide treated mouse eggs manifest similar temporal changes in the decrease in both cdc2/cyclin B kinase and MAP kinase activities that occur following fertilization, although cortical granule exocytosis is not stimulated. The decrease in cdc2/cyclin B kinase activity, however, does not seem to be required for the decrease in MAP kinase activity, since the decrease in MAP kinase activity still occurs in cycloheximide-treated eggs that are also incubated in the presence of nocodazole, which inhibits cyclin B degradation and hence the decrease in cdc2/cyclin B kinase. Following removal of these drugs, cdc2/cyclin B kinase activity remains high, MAP kinase activity increases to levels similar to that in the metaphase II-arrested eggs, and a spindle(s) forms with the chromosomes aligned on a metaphase plate. Results of these experiments suggest that some other protein with a relatively short half-life, e.g. cmos, a known upstream activator of MAP kinase, may be responsible for events leading to the decrease in MAP kinase activity. PMID- 8718667 TI - Hydra regeneration from recombined ectodermal and endodermal tissue. I. Epibolic ectodermal spreading is driven by cell intercalation. AB - Cell-cell interaction and cell rearrangement were examined in the process of epithelial sheet formation during regeneration from hydra cell aggregates. The ectodermal and endodermal epithelial cell layers of Hydra magnipapillata were separated by procaine treatment. Each of the separated layers was then dissociated into single cells and reaggregated to produce ectodermal or endodermal cell aggregates. When the two aggregate types were recombined, a firm adhesion was quickly established between them. This was followed by a vigorous spreading of the ectodermal epithelial cells as a thin layer over the endoderm in a manner similar to the 'epiboly' in some developing embryos. Cell movement in this spreading process was examined using fluorescent dyestaining. It revealed that cells initially located in the inside of the aggregate migrated to intercalate themselves among the cells originally present in the contact surface. This radial cell intercalation took place continuously in the contact surface of both the ectodermal and endodermal aggregates, and produced a rapid growth of the contact surface, eventually leading to complete envelopment of the entire endoderm by the ectoderm. The resulting structure was a small sphere having a two layered epithelial organization as in normal hydra. This sphere regenerated into a complete hydra a few days later. A tryptic extract of hydra membrane fraction specifically inhibited the ectodermal spreading over the endoderm, but not the initial adhesion or the later regeneration processes. These observations suggest that radial cell intercalation at the contact surface plays a crucial role in producing ectodermal spreading and establishing epithelial sheet organization in the recombined aggregates. The intercalation is presumably activated by a signal exchange through the contact surface. The inhibitory effect of the membrane extract suggests that it contains a factor that is involved in some way in this signaling mechanism. PMID- 8718666 TI - The effect of wortmannin on the localisation of lysosomal type I integral membrane glycoproteins suggests a role for phosphoinositide 3-kinase activity in regulating membrane traffic late in the endocytic pathway. AB - Addition of wortmannin to normal rat kidney cells caused a redistribution of the lysosomal type I integral membrane proteins Igp110 and Igp120 to a swollen vacuolar compartment. This compartment did not contain the cation independent mannose 6-phosphate receptor and was depleted in acid hydrolases. It was distinct from another swollen vacuolar compartment containing the cation independent mannose 6-phosphate receptor. The swollen Igp110-positive compartment was accessible to a monoclonal antibody against Igp120 added extracellularly, showing that it had the characteristics of an endosomal compartment. Wortmannin had no gross morphological effect on the trans-Golgi network or lysosomes nor any effect on the delivery to the trans-Golgi network of endocytosed antibodies against the type I membrane protein TGN38. We propose that the observed effects of wortmannin were due to inhibition of membrane traffic between cation independent mannose 6 phosphate receptor-positive late endosomes and the trans-Golgi network and to inhibition of membrane traffic between a novel Igp120-positive, cation independent mannose 6-phosphate receptor-negative late endosomal compartment and lysosomes. The effects of wortmannin suggest a function for a phosphatidylinositol 3-kinase(s) in regulating membrane traffic in the late endocytic pathway. PMID- 8718668 TI - Different distributions of homologous chromosomes in adult human Sertoli cells and in lymphocytes signify nuclear differentiation. AB - Using whole chromosome painting probes for human chromosomes 3,7,8,13,17 and 21 and X and the probe pHY2.1 for the Y chromosome coupled with fluorescent in situ hybridization (FISH) analysis, the distribution of chromosomes is reported in nuclei of Sertoli cells of the adult testis and in stimulated blood lymphocytes. The distribution of chromosomes in the two cell types is significantly different. A strong tendency for each pair of homologues to pair is inferred from the observation of only a single detectable signal in the majority of Sertoli cell nuclei. The sex chromosomes, by contrast, give two clearly separated signals. Interphase nuclei in dividing blood lymphocytes, analysed as controls, also show mainly two separated signals for all non-acrocentric autosomal pairs, but acrocentric pairs no. 13 and 21 show some tendency to associate, probably reflecting satellite association. PMID- 8718669 TI - Plant clathrin heavy chain: sequence analysis and restricted localisation in growing pollen tubes. AB - Clathrin-coated vesicles were isolated from soybean (Glycine max L.) cells in suspension culture and their purity was assessed using SDS-PAGE, peptide sequencing and electron microscopy. Antibodies raised to these coated vesicles were used to immunoscreen a soybean cDNA library in lambda gt11 and isolate a partial clone of the clathrin heavy chain (HC) gene. Full-length cDNA for soybean clathrin HC was deduced by 5' and 3' cDNA amplification. The cDNA encodes an amino acid sequence of 1,700 residues, which is slightly larger than rat clathrin HC and may account for the reduced mobility of plant clathrin on SDS-PAGE. Insertion of these extra residues is largely confined to the amino and carboxy termini. Other domains within the heavy chain arms, including those implicated in light chain binding and trimerisation, are relatively well conserved between eukaryotes. A computer algorithm to determine alpha-helical coiled-coil structures reveals that only one domain, aligning to residues 1,460-1,489 in rat clathrin HC, has a high probability for coiled-coil structure in all five eukaryotic clathrin HC sequences. This provides further evidence that the interaction between clathrin heavy and light chains is mediated by three bundles of coiled-coils near to the carboxy terminus. In analysing the role of plant clathrin in endocytotic trafficking, as against trafficking from the Golgi apparatus to the vacuole, our attention was focused on membrane recycling in tip growing pollen tubes. These rapidly growing cells are highly secretory and require a high level of plasma membrane recycling to maintain the tube tip architecture. Monoclonal antibodies to plant clathrin HC confirmed that coated vesicles are relatively abundant in tip-growing pollen tubes of Lilium longiflorum. This analysis also demonstrated that a high proportion of the clathrin present is in an assembled state, suggesting a highly dynamic trafficking pathway. Immunofluorescence analysis of pollen tubes revealed that clathrin localises to the plasma membrane at the apex of the pollen tube tip, which is consistent with high levels of clathrin-mediated membrane recycling. The use of these reagents in conjunction with tip-growing pollen tubes has created a unique opportunity to examine the basis for constitutive endocytosis, so that the more complex question of receptor-mediated pathways in plants can also be assessed. PMID- 8718670 TI - Partial colocalization of glucocorticoid and mineralocorticoid receptors in discrete compartments in nuclei of rat hippocampus neurons. AB - The glucocorticoid receptor and the mineralocorticoid receptor are hormone dependent transcription factors. They regulate the excitability of rat hippocampus CA1 neurons in a coordinated fashion. We studied the spatial distribution of these transcription factors in nuclei of CA1 neurons by dual labeling immunocytochemistry and confocal microscopy, combined with novel image restoration and image analysis techniques. We found that both receptors are concentrated in about one thousand clusters within the nucleus. Some clusters contain either mineralocorticoid receptors or glucocorticoid receptors, but a significant number of clusters contains both receptors. These results indicate that the two receptor types are targeted to specific compartments in the nucleus. The coordinated action of the glucocorticoid and mineralocorticoid receptor on gene expression may be established in a specific set of nuclear domains that contain both receptors. PMID- 8718671 TI - Direct observation and quantification of macrophage chemoattraction to the growth factor CSF-1. AB - The cloned mouse macrophage cell line, BAC1.25F, resembles primary macrophages in its dependence on colony stimulating factor-1 (CSF-1) for both viability and proliferation. Re-addition of CSF-1 to cytokine-deprived cells, which are rounded with diffusely organised F-actin, stimulates rapid cell spreading and cell polarisation. Using the Dunn chemotaxis chamber the movement of stimulated macrophages was monitored over a 2 hour period. Cells restimulated with 1.32 nM human recombinant CSF-1 migrated at a mean rate of 7.71 microns per hour, but showed no directional preferences. In a linear concentration gradient of CSF-1, cytokine-deprived cells were again stimulated to migrate and the mean rate of cell motility, at 6.88 microns per hour, was not significantly different from that measured in an isotropic environment of CSF-1. However, there was a strong preference for the cells to orientate so that their long axes aligned with the CSF-1 gradient and they migrated preferentially towards the source of CSF-1. Migrating cells contained abundant F-actin within the leading lamellae as judged by confocal imaging of fluorescent phalloidin, but the actin was not arranged into stress fibre-like structures. These data support the proposition that CSF-1 is both a chemokinetic and chemotactic agent for macrophages. Tumour necrosis factor (TNF-alpha) failed to stimulate cell migration and thus was neither chemokinetic nor a chemotactic agent. However, cells exposed to a dual concentration gradient of both TNF-alpha and CSF-1 did migrate successfully, although the chemotactic response to CSF-1 was abolished. PMID- 8718672 TI - A novel calcium-binding protein in amniotic fluid, CAAF1: its molecular cloning and tissue distribution. AB - We found by using a 45Ca2+ overlay technique a large amount of Ca(2+)-binding activity in bovine amniotic fluid from which a novel calcium-binding protein (CaBP) was purified and is referred to as CAAF1 (calcium-binding protein in amniotic fluid-1), with an apparent molecular mass of 8 kDa determined by N tris(hydroxymethyl)-methylglycine/ SDS-PAGE. It was structurally homologous with MRP/calgranulin proteins (MRP8/calgranulin A and MRP14/calgranulin B), members of the S100 protein family, which are abundantly found in the cytoplasm of granulocytes and macrophages. CAAF1 lacked the predicted signal peptide sequence, which is consistent with other CaBPs. The tissue and cellular distribution of CAAF1 was determined by monoclonal antibodies developed against this protein. Its immunoreactivity was found in squamous epithelial cells, neutrophils, and some macrophages throughout the fetal body. An especially characteristic staining pattern was obtained in the squamous epithelium, including that of the esophagus, skin and amnion: CAAF1 was detected in the suprabasal squamous epithelial cells undergoing differentiation, but not in the cells in the proliferating basal layer. Northern blot analysis also showed that CAAF1 mRNA was highly expressed in bovine fetal esophagus and skin. On the other hand, our ELISA studies showed that CAAF1 protein was present in amniotic fluid at a concentration of about 120 nM, which was over 30 times as high as that in the fetal serum. These results suggested that CAAF1 is one of the stage-specific proteins in the differentiation of squamous epithelial cells, and that CAAF1 is preferentially produced by fetal squamous epithelial cells, including epidermal keratinocytes and amniotic epithelial cells, and it is stored in the amniotic fluid during embryogenesis. PMID- 8718673 TI - The relative roles of specific N- and C-terminal phosphorylation sites in the disassembly of intermediate filament in mitotic BHK-21 cells. AB - Previously we identified p34cdc2 as one of two protein kinases mediating the hyperphosphorylation and disassembly of vimentin in mitotic BHK-21 cells. In this paper, we identify the second kinase as a 37 kDa protein. This p37 protein kinase phosphorylates vimentin on two adjacent residues (thr-457 and ser-458) which are located in the C-terminal non-alpha-helical domain. Contrary to the p34cdc2 mediated N-terminal phosphorylation (at ser-55) which can disassemble vimentin intermediate filaments (IF) in vitro, p37 protein kinase phosphorylates vimentin IF without obviously affecting its structure in vitro. We have further examined the in vivo role(s) of vimentin phosphorylation in the disassembly of the IF network in mitotic BHK cells by transient transfection assays. In untransfected BHK cells, the interphase vimentin IF networks are disassembled into non filamentous aggregates when cells enter mitosis. Transfection of cells with vimentin cDNA lacking the p34cdc2 phosphorylation site (ser55:ala) effectively prevents mitotic cells from disassembling their IF. In contrast, apparently normal disassembly takes place in cells transfected with cDNA containing mutated p37 kinase phosphorylation sites (thr457:ala/ser458:ala). Transfection of cells with vimentin cDNAs lacking both the N- and C-terminal phosphorylation sites yields a phenotype indistinguishable from that obtained with the single N terminal mutant. Taken together, our results demonstrate that the site-specific phosphorylation of the N-terminal domain, but not the C-terminal domain of vimentin plays an important role in determining the state of IF polymerization and supramolecular organization in mitotic cells. PMID- 8718674 TI - The cruciated microtubule-associated fibers of the green alga Dunaliella bioculata consist of a 31 kDa SF-assemblin. AB - Cytoskeletons of Dunaliella bioculata, the biflagellate wallless green alga, were isolated and analyzed using a monoclonal and a polyclonal antibody raised against SF-assemblin, the major protein of the two striated microtubule-associated fibers of the alga Spermatozopsis similis. Indirect immunofluorescence showed antigenic structures associated with the four microtubular flagellar roots. SDS-PAGE followed by immunoblot analysis revealed a cross-reacting polypeptide of 31 kDa. This protein of D. bioculata was isolated using gel filtration chromatography in 8 M urea and in vitro reassembly of striated fibers. Microsequencing of the purified protein yielded various peptides, which could be aligned along the sequence of SF-assemblin from S. similis. A complete sequence of the Dunaliella protein was obtained by cDNA cloning. It documents the non helical head domain followed by a helical rod domain with a 29 residue repeat pattern based on four heptads followed by a skip residue. Compared to SF-assemblin of S. similis the SF assemblin of Dunaliella has a shorter head and a slightly longer rod domain. The two algal SF-assemblins share only 57% sequence identity. We conclude that SF assemblin and related proteins in various protists are representatives of a new class of alpha-helical proteins characterized by the ability to form a special segmented coiled coil and to assemble into striated fibers of 2 nm protofilaments in vivo and in vitro. PMID- 8718675 TI - Microtubule-based peroxisome movement. AB - The association of peroxisomes with cytoskeletal structures was investigated both by electron microscopy and by kinetic analysis of peroxisome movement. The morphological studies indicated distinct interactions of peroxisomes with microtubules and frequently revealed multiple contact sites. The kinetic approach utilised microinjection and import of fluorescein-labeled luciferase in order to mark and track peroxisomes in vivo. Peroxisomal motility was analysed by time lapse imaging and fluorescence microscopy. According to their movement peroxisomes were classified into two groups. Group 1 peroxisomes comprising the majority of organelles at 37 degrees C moved slowly with an average velocity of 0.024 +/- 0.012 micron/second whereas the movement of group 2 peroxisomes, 10-15% of the total population, was saltatory exhibiting an average velocity of 0.26 +/- 0.17 micron/second with maximal values of more than 2 microns/second. Saltations were completely abolished by the microtubule-depolymerising drug nocodazole and were slightly reduced by about 25% by cytochalasin D which disrupts the actin microfilament system. Double fluorescence labeling of both peroxisomes and microtubules revealed peroxisome saltations linked to distinct microtubule tracks. Cellular depletion of endogenous levels of NTPs as well as the use of 5' adenylylimidodiphosphate, a nonhydrolysable ATP analog, applied to a permeabilised cell preparation both completely blocked peroxisomal movement. These data suggest an ATPase dependent, microtubule-based mechanism of peroxisome movement. Both the intact and the permeabilised cell system presented in this paper for the first time allow kinetic measurements on peroxisomal motility and thus will be extremely helpful in the biochemical characterisation of the motor proteins involved. PMID- 8718676 TI - sp42, the boar sperm tyrosine kinase, is a male germ cell-specific product with a highly conserved tissue expression extending to other mammalian species. AB - sp42, a tyrosine kinase of 42 kDa originally found in ejaculated boar spermatozoa, is so far the only tyrosine protein kinase to have been purified from mature male germ cells. We have developed and characterized rabbit polyclonal antibodies specifically directed against the boar sperm enzyme, which has been here purified to homogeneity. Anti-sp42 serum and sp42 affinity-purified antibodies work very well in western blot, immunoprecipitation and immunocytochemistry, and do not inhibit sp42 catalytic activity. Immunoblotting analyses reveal the presence of sp42 both in maturing boar epididymal (caput, corpus and cauda segment) spermatozoa and in testicular spermatogenic cells, thus establishing that the protein is effectively expressed in the germ cells and is not a sperm-associated protein secreted by the epididymal epithelium or male accessory glands. This finding is further strengthened by the fact that sp42 is not glycosylated, since different lectins fail to bind to sp42 and treatment of sp42 with different deglycosylation enzymes does not result in a reduction of the molecular mass of sp42. When different boar tissues are immunoscreened in western blot analysis, the results are all sp42-negative. The extension of the study to other mammalian species (human, mouse and rat) demonstrates that proteins immunologically related to boar sp42, which share the same molecular mass and tyrosine kinase activity, are both expressed in spermatogenic cells and maintained in mature sperm cells. Intriguingly, when a wide spectrum of somatic mouse and rat tissues is probed with sp42-antiserum, no tissue presents anti-sp42 immunoreactivity. Immunocytochemistry shows that in boar spermatozoa sp42 is confined to the tail mid-piece, while by immunohistochemistry carried out on sections of adult rat testis the appearance time of the kinase appears to be consistent with a post-meiotic synthesis in haploid spermatids. Altogether, these results demonstrate that boar sp42 is a new male germ cell-specific gene product, with highly conserved tissue expression extended to other mammalian species, and suggest a possible role played by the cytoplasmic tyrosine kinase in the cell signalling network specific to haploid male germ cells. PMID- 8718677 TI - rRNA distribution during microspore development in anthers of Beta vulgaris L. quantitative in situ hybridization analysis. AB - We related changes in the ultrastructural organization of the nucleoli with the results of quantitative in situ hybridizations to characterize rRNA metabolism during the development of microspore mother cells in the sugar beet anther (Beta vulgaris L.). In the course of meiotic prophase and early postmeiotic interphase the morphological characteristics of the nucleoli are typical of low or no transcriptional activity and a low rate of rRNA processing. However, we found evidence of an apparent increase in the relative numbers of 18 S rRNA transcripts in some stages of microsporogenesis. This was found in both the nucleoli and cytoplasm of pachytene meiocytes, and in later stages there was a spectacular accumulation of rRNA transcripts in nucleoli of the tetrad cells. Quantitative data are analyzed in the light of morphometric findings in the cell and their compartments to elucidate the degree to which changes in cell size are related to changes in labeling density and distribution. The results are discussed in terms of rRNA synthesis, transport and degradation as processes involved in the regulation of rRNA within microsporocytes and microspores. PMID- 8718678 TI - An ultradian clock controls locomotor behaviour and cell division in isolated cells of Paramecium tetraurelia. AB - An ultradian clock operates in fast growing cells of the large ciliate, Paramecium tetraurelia. The period of around 70 minutes is well temperature compensated over the temperature range tested, i.e. between 18 degrees C and 33 degrees C. The Q10 between 18 degrees C and 27 degrees C is 1.08; above 27 degrees C there is a slight overcompensation. The investigation of individual cells has revealed that two different cellular functions are under temporal control by this ultradian clock. First, locomotor behaviour, which is an alternation between a phase of fast swimming with only infrequent turning, and a phase of slow swimming with frequent spontaneous changes of direction. In addition, the ultradian clock is involved in the timing of cell division. Generation times are not randomly distributed, but occur in well separated clusters. At all of the six temperatures tested, the clusters are separated by around 70 minutes which corresponds well to the period of the locomotor behaviour rhythm at the respective temperatures. Whereas the interdivision times were gradually lengthened both above and below the optimum growth temperature, the underlying periodicity remained unaffected. Also cells of different clonal age had identical periods, suggesting that neither the differences in DNA content, not other changes associated with ageing in Paramecium have an effect on the clock. A constant phase relationship was observed between the rhythm in locomotor behaviour and the time window for cell division; this strongly suggests that the same ultradian clock exerts temporal control over both processes. PMID- 8718679 TI - gamma-Tubulin in mammalian cells: the centrosomal and the cytosolic forms. AB - The centrosome is one of the cellular organelles for which the mechanism by which it operates still remains to be unlavelled. The finding of the association with the centrosome of gamma-tubulin, a protein which belongs to the tubulin superfamily, has provided a long sought after biochemical tool with which to address centrosome function. We have generated a specific anti-gamma-tubulin polyclonal antibody to study the biochemical properties and the cellular distribution of the human lymphoblastic gamma-tubulin. Using cell fractionation and mass isolation of centrosomes, we observed that in contrast to the figures suggested by immunofluorescence, a minimum figure of 80% of total gamma-tubulin exists as a cytosolic form. The centrosomal form, for which at least half is not strongly associated with the centrosome, behaves in two-dimensional gel electrophoresis identically to the soluble form (as at least two spots of a pI of around 6). Post-embedding immunolocalization reveals that gamma-tubulin is distributed in the pericentriolar matrix but is also closely associated with centrioles. Using a combination of gel filtration, ion exchange chromatography, equilibrium sucrose gradient centrifugation and immunoprecipitation, we show that the major part of cytosolic gamma-tubulin might be involved in complexes heavier than the Tcp1 particle. We further demonstrate, by co-immunoprecipitation of gamma-tubulin and Tcp1 with either anti-Tcp1 or anti-gamma-tubulin antibodies, that a small part of gamma-tubulin participates in Tcp1-gamma-tubulin particles. Interestingly, the soluble form of gamma-tubulin co-purifies with taxol stabilized microtubules and its association with microtubules resisted salt, ATP and GTP treatments. The existence of a centrosomal form and a large pool of cytosolic gamma-tubulin-containing complexes in somatic cells suggests that the overall gamma-tubulin cellular distribution does not seem to be as straightforward as it was drawn earlier. PMID- 8718680 TI - Localization of kinesin superfamily proteins to the connecting cilium of fish photoreceptors. AB - Kinesin superfamily proteins (KIFs) are probable motors in vesicular and non vesicular transport along microtubular tracks. Since a variety of KIFs have been recently identified in the motile flagella of Chlamydomonas, we sought to ascertain whether KIFs are also associated with the connecting cilia of vertebrate rod photoreceptors. As the only structural link between the rod inner segment and the photosensitive rod outer segment, the connecting cilium is thought to be the channel through which all material passes into and out of the outer segment from the rod cell body. We have performed immunological tests on isolated sunfish rod inner-outer segments (RIS-ROS) using two antibodies that recognize the conserved motor domain of numerous KIFs (anti-LAGSE, a peptide antibody, and anti-Klp1 head, generated against the N terminus of Chlamydomonas Klp1) as well as an antibody specific to a neuronal KIF, KIF3A. On immunoblots of RIS-ROS, LAGSE antibody detected a prominent band at approximately 117 kDa, which is likely to be kinesin heavy chain, and Klp1 head antibody detected a single band at approximately 170 kDa; KIF3A antibody detected a polypeptide at approximately 85 kDa which co-migrated with mammalian KIF3A and displayed ATP dependent release from rod cytoskeletons. Immunofluorescence localizations with anti-LAGSE and anti-Klp1 head antibodies detected epitopes in the axoneme and ellipsoid, and immunoelectron microscopy with the LAGSE antibody showed that the connecting cilium region was particularly antigenic. Immunofluorescence with anti KIF3A showed prominent labelling of the connecting cilium and the area surrounding its basal body; the outer segment axoneme and parts of the inner segment coincident with microtubules were also labelled. We propose that these putative kinesin superfamily proteins may be involved in the translocation of material between the rod inner and outer segments. PMID- 8718682 TI - Specific cleavage of target RNAs from HIV-1 with 5' half tRNA by mammalian tRNA 3' processing endoribonuclease. AB - Mammalian tRNA 3' processing endoribonuclease (3' tRNase) can be converted to an RNA cutter that recognizes four bases, with about a 65-nt 3'-truncated tRNA(Arg) or tRNA(Ala). The 3'-truncated tRNA recognizes the target RNA via four base pairings between the 5'terminal sequence and a sequence 1-nt upstream of the cleavage site, resulting in a pre-tRNA-like complex (Nashimoto M, 1995, Nucleic Acids Res 23:3642-3647). Here I developed a general method for more specific RNA cleavage using 3' tRNase. In the presence of a 36-nt 5' half tRNA(Arg) truncated after the anticodon, 3' tRNase cleaved the remaining 56-nt 3' half tRNA(Arg) with a 19-nt 3' trailer after the discriminator. This enzyme also cleaved its derivatives with a 5' extra sequence or nucleotide changes or deletions in the T stem-loop and extra loop regions, although the cleavage efficiency decreases as the degree of structural change increases. This suggests that any target RNA can be cleaved site-specifically by 3'tRNase in the presence of a 5' half tRNA modified to form a pre-tRNA-like complex with the target. Using this method, two partial HIV-1 RNA targets were cleaved site-specifically in vitro. These results also indicate that the sequence and structure of the T stem-loop domain are important, but not essential, for the recognition of pre-tRNAs by 3' tRNase. PMID- 8718681 TI - Intramolecular structure in yeast introns aids the early steps of in vitro spliceosome assembly. AB - rp5l B pre-mRNA, like many Saccharomyces cerevisiae primary transcripts, contains a secondary structure within its intron sequence. The structure is required for optimal in vivo splicing efficiency and includes two complementary regions near the 5' splice site and the branchpoint (UB1 and DB1, respectively). An intron containing RNA was probed in vitro with RNase T1 and dimethyl sulfate (DMS), and is folded as expected. We have also examined in vitro splicing of rp5l B pre mRNA, by analyzing the formation of splicing complexes as well as splicing products. Similar analyses were done with mutant pre-mRNAs containing shortened or lengthened UB1/DB1 base pairing regions. Our experiments indicate that the secondary structure acts at an early step of spliceosome assembly to aid the formation of U1 snRNP-containing commitment complexes. pre-mRNAs were probed with DMS in vivo and the folding takes place inside cells. The effect of the different UB1/DB1 interactions on in vivo splicing efficiency was also analyzed. The results are consistent with the idea that the intramolecular interaction takes place prior to or at the beginning of spliceosome assembly. PMID- 8718683 TI - Essential domains of the PRP21 splicing factor are implicated in the binding to PRP9 and PRP11 proteins and are conserved through evolution. AB - The yeast Prp9p, Prp11p, Prp21p proteins form a multimolecular complex identified as the SF3a splicing factor in higher eukaryotes. This factor is required for the assembly of the prespliceosome. Prp21p interacts with both Prp9p and Prp11p, but the molecular basis of these interactions is unknown. Prp21p, its human homologue, and the so-called SWAP proteins share a tandemly repeated motif, the surp module. Given the evolutionary conservation and the role of SWAP proteins as splicing regulators, it has been proposed that surp motifs are essential for interactions between Prp21p and other splicing factors. In order to characterize functional domains of Prp21p and to identify potential additional functions of this protein, we isolated a series of heat-sensitive prp21 mutants. Our results indicate that prp21 heat-sensitive mutations are associated with defects in the interaction with Prp9p, but not with Prp11p. Interestingly, most heat-sensitive point mutants associate a strong splicing defect with a pre-mRNA nuclear export phenotype, as does the prp9-1 heat-sensitive mutant. Deletion analyses led to the definition of domains required for viability. These domains are responsible for the interaction with Prp9p and Prp11p and are conserved through evolution. They do not include the most conserved surp1 module, suggesting that the conservation of this motif in two families of proteins may reflect a still unknown function dispensable in yeast under standard conditions. PMID- 8718684 TI - Domain structure of the ribozyme from eubacterial ribonuclease P. AB - Large RNAs can be composed of discrete domains that fold independently. One such "folding domain" has been identified previously in the ribozyme from Bacillus subtilis ribonuclease P (denoted P RNA). This domain contains roughly one-third of all residues. Folding of an RNA construct consisting of the remaining two thirds of B. subtilis P RNA was examined by Fe(II)-EDTA hydroxyl radical protection. This molecule folds into the proper higher-order structure under identical conditions as the full-length P RNA, suggesting the presence of a second folding domain in B. subtilis P RNA. Folding analysis of the Escherichia coli P RNA by hydroxyl radical protection shows that this P RNA is completely folded at 5-6 mM Mg2+. In order to analyze the structural organization of folding domains in E. coli P RNA, constructs were designed based on the domain structure of B. subtilis P RNA. Fe(II)-EDTA protection indicates that E. coli P RNA also contains two folding domains. Despite the significant differences at the secondary structure level, both P RNAs appear to converge structurally at the folding domain level. The pre-tRNA substrate, localized in previous studies, may bind across the folding domains with the acceptor stem/3'CCA contacting the domain including the active site and the T stem-loop contacting the other. Because all eubacterial P RNAs share considerable homology in secondary structure to either B. subtilis or E. coli P RNA, these results suggest that this domain structure may be applicable for most, if not all, eubacterial P RNAs. Identification of folding domains should be valuable in dissecting structure function relationship of large RNAs. PMID- 8718685 TI - Slow folding kinetics of RNase P RNA. AB - Understanding the folding mechanisms of large, highly structured RNAs is important for understanding how these molecules carry out their function. Although models for the three-dimensional architecture of several large RNAs have been constructed, the process by which these structures are formed is only now beginning to be explored. The kinetic folding pathway of the Tetrahymena ribozyme involves multiple intermediates and both Mg2+-dependent and Mg2+-independent steps. To determine whether this general mechanism is representative of folding of other large RNAs, a study of RNase P RNA folding was undertaken. We show, using a kinetic oligonucleotide hybridization assay, that there is at least one slow step on the folding pathway of RNase P RNA, resulting in conformational changes in the P7 helix region on the minute timescale. Although this folding event requires the presence of Mg2+, the slow step itself does not involve Mg2+ binding. The P7 and P2 helix regions exhibit distinctly different folding behavior and ion dependence, implying that RNase P folding is likely to be a complex process. Furthermore, there are distinct similarities in the folding of RNase P RNA from both Bacillus subtilis and Escherichia coli, indicating that the folding pathway may also be conserved along with the final structure. The slow folding kinetics, Mg2+-independence of the rate, and existence of intermediates are basic features of the folding mechanism of the Tetrahymena group I intron that are also found in RNase P RNA, suggesting these may be general features of the folding of large RNAs. PMID- 8718686 TI - Conditional inactivation of mRNA capping enzyme affects yeast pre-mRNA splicing in vivo. AB - Acquisition of the 5'cap is the earliest modification event during eukaryotic mRNA synthesis. The cap is thought to facilitate later processing steps, such as pre-mRNA splicing. If this is so, then a defect in cap synthesis should impact on splicing in vivo. We tested this hypothesis by examining the consequences of conditional inactivation of the Saccharomyces cerevisiae CEG1 gene, which encodes mRNA guanylyltransferase (capping enzyme). Two different ceg1-ts mutants, Y66A and C354Y, displayed a pre-mRNA processing (prp) defect, characterized by an increase in the amount of unspliced pre-mRNA after shift to nonpermissive temperature and a decrease in the amount of mature mRNA. The guanylyltransferase activities of the Y66A and C354Y proteins were thermolabile, suggesting that defective capping in vivo was contributory to the prp phenotype. Although these results provide the first genetic link between capping and splicing in vivo, we were unable to demonstrate a role for either the cap or the capping enzyme during yeast pre-mRNA splicing in vitro. PMID- 8718687 TI - Conditional mutants of the yeast mRNA capping enzyme show that the cap enhances, but is not required for, mRNA splicing. AB - The 5' end of eukaryotic mRNAs are modified by the addition of a 7-methyl guanosine (m7G) cap. The role of the cap in translation has been well established. Additionally, studies conducted in vitro or in microinjected Xenopus oocytes have implicated the cap in RNA processing and transport. To determine the fate of uncapped mRNA in intact yeast cells, conditional alleles of the gene encoding the capping enzyme guanylyltransferase subunit (CEG1) were generated. RNA analysis of temperature-sensitive ceg1 strains revealed an accumulation of unspliced pre-mRNAs and a corresponding decrease in spliced mRNAs at the restrictive temperature. A substantial proportion of spliced mRNA was also uncapped. Therefore, the cap appears to stimulate, but is not absolutely required for, splicing in vivo. In addition, steady-state levels of several mRNAs were decreased, perhaps due to increased degradation of uncapped mRNAs. In contrast to splicing, mRNA polyadenylation and transport to the cytoplasm were unaffected. PMID- 8718688 TI - Directed hydroxyl radical probing of the rRNA neighborhood of ribosomal protein S13 using tethered Fe(II). AB - Directed hydroxyl radical probing was used to probe the rRNA neighborhood around protein S13 in the 30S ribosomal subunit. The unique cysteine at position 84 of S13 served as a tethering site for attachment of Fe(II)-1-(p bromoacetamidobenzyl)-EDTA. Derivatized S13 (Fe-C84-S13) was then assembled into 30S ribosomal subunits by in vitro reconstitution with 16S rRNA and a mixture of the remaining 30S subunit proteins. Hydroxyl radicals generated from the tethered Fe(II) resulted in cleavage of the RNA backbone in two localized regions of the 3' major domain of 16S rRNA. One region spans nt 1308-1333 and is close to a site previously crosslinked to S13. A second set of cleavages is found in the 950/1230 helix. Both regions have been implicated in binding of S13 by previous chemical footprinting studies using base-specific chemical probes and solution-based hydroxyl radical probing. These results place both regions of 16S rRNA in proximity to position C84 of S13 in the three-dimensional structure of the 30S ribosomal subunit. PMID- 8718689 TI - Exon circularization in mammalian nuclear extracts. AB - Correct ligation of exons in pre-mRNA splicing requires splice site juxtaposition (splice site pairing), usually involving a 5' splice site and a downstream 3' splice site. Splicing of a 5' splice site to an upstream 3' splice site, however, is predicted to result in a circular RNA. This mode of splice site pairing across the axon has been hypothesized to account for rare RNAs containing scrambled exons (Nigro JM et al., 1991, Celt 64:607-613; Cocquerelle C et al., 1992, EMBO J 11:1 095-1098). Additionally, this mode of splice site pairing has been postulated to explain the formation of SRY circular transcripts in mouse testis (Capel B et al., 1993, Celt 73:1019- 1030). Here we show that splice site pairing across the exon can result in exon circularization in vitro. These results indicate that spliceosome-mediated axon circularization indeed can account for the formation of scrambled exons and circular RNAs. Exon circularization efficiency decreased dramatically as the length of the exon was increased from 95 nt to 274 nt. Circularization of this longer exon was restored, however, when intronic complementary sequences were included in the RNA substrate. These complementary sequences could form a stem that served to bring the splice sites into proximity and thereby promote splice site pairing. Therefore, the splicing of this structured RNA recapitulated SRY-like exon circularization in vitro. PMID- 8718690 TI - Detection of double-stranded RNA-protein interactions by methylene blue-mediated photo-crosslinking. AB - Double-stranded(ds) RNA-binding proteins have diverse functions in the cell. An obstacle to investigating the interactions between these proteins and dsRNA is the relative inefficiency of traditional UV-crosslinking methods for extended regions of dsRNA. We have therefore developed an alternative procedure for RNA protein photo-crosslinking that efficiently induces RNA-protein crosslinks in double-stranded regions of RNA. We show that dsRNA-protein crosslinks can be induced by visible light in the presence of the dye methylene blue, which most likely mediates crosslinking by intercalating in the dsRNA helix. A recombinant dsRNA binding domain from the Drosophila staufen protein and human protein kinase R were crosslinked by UV or methylene blue to a series of dsRNAs. In each case, the degree of crosslinking was greater with methylene blue, particularly with RNAs with few single-stranded loops. Methylene blue-mediated crosslinking therefore complements and extends the existing repertoire of crosslinking methods for detecting RNA-protein interactions. PMID- 8718692 TI - Identification of a new high oxygen affinity hemoglobin variant: Hb Aurora [beta 139(H17) Asn-->Tyr]. AB - A 73-year-old female of Dutch descent was referred for investigation of a high oxygen affinity hemoglobin variant. The beta-globin gene was amplified using the polymerase chain reaction. Direct nucleotide sequencing of the polymerase chain reaction amplified DNA revealed that she is heterozygous for a novel beta-globin gene mutation at codon 139, AAT-->TAT. The resulting hemoglobin variant has been designated Hb Aurora [beta 139 (H17) Asn-->Tyr]. PMID- 8718691 TI - The sickle cell painful crisis in adults: phases and objective signs. AB - The severe sickle cell painful crisis that requires hospitalization in adults typically seems to evolve along four distinct phases: Prodromal, initial, established, and resolving. Each phase may to be associated with certain clinical and laboratory findings. Objective clinical and laboratory signs may not be evident in the initial phase of the crisis especially in the absence of reliable base line steady state data. Several clinical, hematological, biochemical, and rheological changes emerge as the crisis progresses. This sequence of events creates several logistic issues in the care of these patients. The absence of objective findings early in the crisis usually discourages health care providers from repeating such tests and makes some of them suspicious of the authenticity of the painful episode in question. Consequently, an atmosphere of mistrust may develop between the patients and their care providers. This review puts the sickle cell painful crisis in perspective and, based on available data in the literature, shows that it is a dynamic process characterized by several changes during its evolution. It is hoped that this review will dispel some misconceptions about the painful crisis, elucidate its clinical course, and encourage further research to answer the questions it raises. PMID- 8718693 TI - Molecular basis of alpha-thalassemia in Portugal. AB - We have estimated the incidence and molecular basis of alpha-thalassemia in a Portuguese population, mostly from the Greater Lisbon area. In a group of 100 consecutive cord blood samples, the gene frequency of the rightward deletion ( alpha 3.7) was 0.035, and the leftward deletion (-alpha 4.2) was 0.015. In this group, we have also found four heterozygotes for the triple alpha-globin gene rearrangement (alpha alpha alpha anti 3.7. gene frequency 0.020). We have characterized the subtypes of -alpha 3.7 and alpha alpha alpha anti 3.7 rearrangements. On the whole, these results give an incidence of 10% for deletional alpha-thalassemia carriers in the studied Portuguese population. In a group of 342 subjects presenting beta-thalassemia, or Hb S trait, beta thalassemia major sickle cell disease or low red blood cell indices, the -alpha 3.7, -alpha 4.2, -SEA, -MED, (alpha alpha)MM, and alpha alpha alpha anti 3.7 haplotypes were found in different combinations. Only one nondeletional alpha thalassemia determinant (a 5 nucleotide deletion in the alpha 2-globin gene in the second intervening sequence donor site) was detected, which might suggest a low incidence of these defects in the Portuguese population. PMID- 8718694 TI - The frequency of 14 beta-thalassemia mutations in the Arab populations. AB - The beta-thalassemias are a heterogeneous group with respect to molecular pathogenesis, and different populations and ethnic groups differ with respect to the predominating mutations. This variable spectrum of beta-thalassemia mutations has resulted in extensive studies in each population and ethnic group to identify the major mutations. In this study we investigated the prevalence of 14 mutations in 253 beta-thalassemia patients drawn from eight Arab countries (i.e. Jordan, Egypt, Syria, Lebanon, Yemen, and Saudi Arabia), living in Saudi Arabia and attending Ministry of Health hospitals. The mutations investigated included IVS-I 110 (G-->A), IVS-II-1 (G-->A), IVS-I-5 (G-->C), codon 39 (C-->T), IVS-I-1 (G- >A), frameshift at codons 8/9 (+G), frameshift at codons 41/42 (-TTCT), codon 15 (TGG-->TAG), IVS-I-6 (T-->C), frameshift at codon 16 (-C), IVS-II-745 (C-->G), codon 6(-A), IVS-I, 3' end (-25 bp), and Cap +1 (A-->C). The most frequently encountered mutations were IVS-I-110 and IVS-II-1 which were identified in the population of each Arab country. The IVS-I-1 and IVS-II-745 mutations were encountered in Jordanians, Egyptians, and Syrians. The IVS-I-5, codon 39, codon 6, IVS-I, 3' end (-25 bp), and Cap +1 mutations were encountered only in Saudis and not in other Arabs, except codon 39 which was present in the Syrians and Lebanese. Other mutations were generally rare and not specific to any Arab ethnic group. This paper presents preliminary data on the prevalence of 14 mutations in the Arab populations and shows wide variation in the molecular basis of beta thalassemia in different Arab ethnic groups. Further detailed studies to identify the entire spectrum of beta-thalassemia mutations are stressed. PMID- 8718695 TI - Structural characterization of the beta-globin gene cluster in an individual expressing a very low level of G gamma globin chains. AB - We report the case of a normal individual displaying an extremely unbalanced G gamma/A gamma-globin ratio (G gamma-globin chains undetectable by urea/triton/ acrylamide gel electrophoresis and just reaching the threshold of detection by high performance liquid chromatography) associated with a very low level of G gamma-globin mRNA (at the most 5% of total gamma-mRNA after reverse transcriptase polymerase chain reaction determination). By DNA Southern blotting and sequencing, the very low level of G gamma-globin chains in this individual was found in association with subhaplotype [+ -----] (Hinc II 5' to epsilon, Xmn I 5' to G gamma, Hind III in G gamma and A gamma, Hinc II in and 3' to psi beta), with G gamma- and A gamma-globin gene sequences of the B type chromosome, and with a number of AT repeats in the locus control region hypersensitive site-2 site, similar to that reported to be associated with the Bantu beta S haplotype. These structural characteristics, described for the first time combined in the same individual, suggest that the G gamma/A gamma ratio in adults, is controlled by sequences distributed all along the beta-globin gene cluster. PMID- 8718696 TI - A novel intrachromosomal rearrangement in the beta-globin gene found in an African-American family. AB - We describe here a deletion of 34 nucleotides from the 3' end of the first intervening sequence of the beta-globin gene covering the AGGC splice junction, and the insertion of 32 nucleotides of the delta-globin gene at the same location. This gene rearrangement was detected in three members of an African American family. The proband, a 28-year-old female, and her mother had a history of chronic anemia. One of her two brothers, who inherited the same gene defect, was apparently healthy with no symptoms of hemolytic anemia. The proband, her father, and her two brothers, including the one who carried the beta-globin gene rearrangement, were found to be heterozygous for alpha-thalassemia-2 (-alpha 3.7). Although the AGGC splice junction is disrupted (AGGC-->AGAT), the invariant AG has remained intact after this gene rearrangement. Our investigations could not detect any defect in RNA processing in the affected beta-globin genes. The discrepancies between the phenotypes and the globin chain synthesis ratios of the mother, her daughter, and her son who inherited the same gene defect at their beta-globin genes, remain unexplained. PMID- 8718698 TI - Hb Nakano [beta 8(Lys-->Met]: a new beta chain variant found in a Japanese woman. PMID- 8718697 TI - Two new alpha chain variants: Hb Fuchu-I [alpha 72(EF1)His-->Tyr] and Hb Fuchu-II [alpha 97(G4)Asn-->His]. PMID- 8718699 TI - The first case of Hb E-Saskatoon [alpha 2 beta(2)22(B4)Glu-->Lys] in a Japanese male in Asia. PMID- 8718700 TI - Hb F-Veleta or alpha 2 G gamma(2)40(C6)Arg-->Gly. PMID- 8718701 TI - The G gamma T chain (G gamma 75 Thr; 136 Gly) in Hb F-Charlotte is the product of an A gamma gene with a limited gene conversion and that in Hb F-Waynesboro of a mutated G gamma gene. PMID- 8718702 TI - A novel DNA polymorphism of the human A gamma-globin gene (A gamma-588, A-->G) is linked with the XMN / polymorphism (G gamma-158, C-->T). PMID- 8718703 TI - Molecular analyses of beta-thalassemia in Iran. PMID- 8718704 TI - Common addictions. PMID- 8718705 TI - Coupled Markov chain model: characterization of membrane channel currents with multiple conductance sublevels as partially coupled elementary pores. AB - A parameterized Markov chain model is developed to represent the characteristics of channel currents that either are the superposition of many single channels or show multiple conductance sublevels. The simplified model takes the form of a set of binary chains that are interdependent according to a simple lumped coupling parameter. When varied, this parameter realizes a range of behaviors from tight coupling to complete independence. Other model parameters describe the intrinsic characteristics of the binary chains. An identification procedure for the model parameters is developed based on hidden Markov modeling ideas but incorporating a novel parameter estimation. The usefulness of the model in analyzing certain types of data is demonstrated with examples of real channel currents. PMID- 8718706 TI - A control problem in a polluted environment. AB - Due to the importance of remediating polluted environments, many mathematical models have been developed to describe population-toxicant interactions. This article considers the situation where the amount of pollution in the environment is restricted to lie below a certain value M. A model is presented and viewed as a control problem in which the toxicant input into the environment is the controllable variable. An explicit formula is computed for the input function that meets certain restrictions. By analytically studying the dynamics of the population, it is possible to provide conditions on how to set M in order to guarantee survival of the population without a significant reduction of its carrying capacity. Numerical and experimental data are described that support these results. PMID- 8718707 TI - Measures of concurrency in networks and the spread of infectious disease. AB - An investigation is made into the impact of concurrent partnerships on epidemic spread. Starting from a definition of concurrency on the level of individuals, the authors define ways to quantify concurrency on the population level. An index of concurrency based on graph theoretical considerations is introduced, and the way in which it is related to the degree distribution of the contact graph is demonstrated. Then the spread of an infectious disease on a dynamic partnership network is investigated. The model is based on a stochastic process of pair formation and separation and a process of disease transmission within partnerships of susceptible and infected individuals. Using Monte Carlo simulation, the spread of the epidemic is compared for contact patterns ranging from serial monogamy to situations where individuals can have many partners simultaneously. It is found that for a fixed mean number of partners per individual the distribution of these partnerships over the population has a major influence on the speed of the epidemic in its initial phase and consequently in the number of individuals who are infected after a certain time period. PMID- 8718708 TI - Estimation of certain parameters of a stationary hybrid process involving a time series and a point process. AB - A method is presented for estimating the cross-spectral density of a hybrid process involving a time series and a point process. The method is based on the generalized cross-periodogram statistic, which is smoothed by splitting the whole record of the data into a number of disjoint subrecords. Estimates of the coherence function and the cross-covariance function can also be obtained by using the estimate of the cross-spectral density. The distribution of the cross covariance function between a time series and a point process is shown to be asymptotically normal. The theoretical results are used in the study of a complex physiological system. It is shown that the presence of a gamma motor neuron (gamma stimulation) modifies the effect of the length changes on the complex system at low frequencies (the length changes, and the response of the system become uncorrelated in the range 3-30 Hz) while the effect remains unchanged at higher frequencies. As a comparison it is shown that the presence of the length changes weakens the effect of the gamma stimulation on the complex system. PMID- 8718710 TI - Saving our cities. PMID- 8718709 TI - Urban mortality--race or place? AB - Several concomitant trends have occurred in American society in the 20th century. First, life expectancy has improved overall, and the gap between blacks and whites has narrowed. Second, as the nature of the economy has changed from rural agrarian to urban postindustrial, there have been fundamental changes in population residential patterns, with most Americans now living in metropolitan areas. Within metropolitan areas, blacks have become concentrated in poor inner cities as whites have moved to the more affluent suburbs. Black mortality rates are higher in big cities than elsewhere, and appear to be directly related to the proportion of blacks in those cities. Black-white mortality ratios, however, are lower in cities of medium size than in either very large or small cities. At the national level age-adjusted mortality ratios between blacks and whites are associated with different causes of death; but only limited cause-specific mortality data are available for large cities. Understanding and ameliorating social conditions that lead to elevated black mortality in U.S. cities will require more information at the municipal level than is currently available. PMID- 8718711 TI - Occupational injury fatalities--1994. AB - Factory workers caught in machinery and construction workers falling or struck by huge beams are images that typically come to mind when considering serious hazards in the workplace. But these types of events account for only a small portion of job-related fatalities each year. Transportation-related fatalities, along with assaults and violent acts during work, made up almost two-thirds of the 6,588 fatal work injuries recorded in 1994. The majority of job-related fatal work events occurred on the streets and highways and in public buildings and in areas such as grocery stores and parking lots. Today the most deadly jobs are found in outdoor occupations such as fishing and timber cutting. In fact, in all 10 jobs studied that have high fatality rates, most workers are affected by severe weather conditions while driving on highways, performing farm chores and working at construction sites. Highway crashes are the primary cause of trucker fatalities; falls are the leading cause of death for roofers, construction laborers and structural metal workers, while tractor rollovers account for a third of farm worker fatalities. Another deadly contributing factor for some workers is homicide, which accounted for 16 percent of job-related fatalities in 1994. Workers most at risk are those who work alone, work late at night and handle varying sums of money. Taxicab drivers are the most susceptible and have a work injury fatality rate nine times higher than the national rate of 5 deaths per 100,000 workers. Others at high risk of homicide include gas station cashiers, grocery store employees and workers in retail eating and drinking establishments. Although the risk of a fatal injury at work varies greatly by occupation and industry, no one is immune. For prevention, workers and employers need to know what jobs are risky, what equipment is dangerous and what activities are hazardous. They also should understand that a fatal incident can happen to anyone. PMID- 8718712 TI - Average charges for a total knee replacement: United States, 1994. AB - Total knee replacements (TKRs) have increased more than fourfold since 1989. The charges for these surgeries have been increasing also, and in 1994 claims to MetLife for a TKR averaged $28,340. Three geographic areas reported average total charges above this norm and six below. The highest average total charge (including hospital and physician charges) was reported in New York and California, each with charges 27.1 percent above the U.S. average. These charges are contrasted with the low average charge reported in Ohio, 19.3 percent below the norm. About one-fourth of the total charges were for physician fees, which averaged $7,150 across the country. Among the 16 study states in which at least 15 TKRs were performed in 1994, the average totals ranged from $11,930 in New York to $6,290 in Colorado. On average, the patients remained hospitalized for 6.7 days, with major state variations noted. The longest stay was reported in New York (more than 11.5 days) and the shortest in Arizona (4.6 days). Per diem charges generally were inversely related to the hospital stays, averaging $4,230 and ranged from a high of $6,640 in Arizona to $3,110 in New York. PMID- 8718713 TI - Geographic changes in state population growth. AB - The population of the United States continues to increase, albeit at a slower pace than in the past. Geographically, the states of the South and West still are experiencing larger relative gains than those in the Northeast and Midwest, although some changes in the decennial growth pattern have recently emerged. California's population increments have steadily diminished since 1990, and last year Florida's relative gain was comparatively small. Moreover, New York recorded a decline in the number of its residents. In contrast, the count of persons living in Texas appears to be accelerating. Together, the population of these four states comprises nearly one-third of the nation's total and, therefore, the United States' future will be heavily influenced by the changes in their growth patterns. PMID- 8718714 TI - Intrapatellar fat pad. PMID- 8718715 TI - The effect of bone quality on the outcome of ingrowth total knee arthroplasty. AB - Bone quality generally is felt to be an important consideration when planning total knee arthroplasty (TKA). This prospective study was conducted to see if bone quality, as assessed intraoperatively in 346 knees, affects the outcome of ingrowth TKA. Group A consisted of 272 knees with good or excellent bone quality in all areas and Group B consisted of 74 knees with fair or poor bone in at least one area. The average follow-up was 48 months, with 301 knees (87%) available for follow-up. Group A knees had more males, were heavier, had fewer patients with rheumatoid arthritis, and exhibited more varus. Preoperatively, Groups A and B had modified Hospital for Special Surgery (HSS) knee scores that averaged 55 and 48, respectively. At 4 years post-surgery, Groups A and B had HSS scores of 92 and 90, respectively. Fluoroscopic radiographs of the bone-prosthesis interfaces showed 13% incomplete radiolucencies in Group A and 7% in Group B. A complete radiolucency was seen in one patient in Group A. Based on these results, it can be concluded that bone quality as determined by gross intraoperative inspection had little effect on the 4-year outcome of this ingrowth TKA. PMID- 8718716 TI - Impacted cancellous autograft for contained bone defects in total knee arthroplasty. AB - Techniques of bone grafting for peripheral tibial defects in total knee arthroplasty are well described using solid bone graft or metal wedges. Central contained defects may be irregularly shaped and not easily managed with bulk grafts or wedges. Morselized grafts loosely packed into the defect may allow cement to intrude into the interstices between the graft fragments. However, impaction of the morselized autograft into a contained defect can permit complete filling of the area with graft and prevent undesirable cement intrusion. This article describes the successful use of this technique in eight patients who underwent primary total knee arthroplasty. PMID- 8718717 TI - The fat pad. Clinical observations. AB - This article retrospectively reviews 53 patients with various knee joint conditions in which the fat pad was implicated. The patients were divided into four groups: patellar tendonitis, cruciate ligament surgery, meniscal surgery, and miscellaneous. Patient ages ranged from 17 to 67 years, and follow-up ranged from 4 weeks to 5 years. All of the patients presented with pain or stiffness that was related in part to the fat pad pathology. This finding was confirmed by an injection of local anesthetic and steroid into the fat pad itself, which was followed by transient although complete symptomatic relief. We conclude that fat pad pathology is usually secondary to other knee joint pathology, and its primary involvement is rare. The pain produced by the fat pad may have serious implications on knee joint function if not dealt with promptly. Appreciation of the source of pain in knee joint pathology will allow better management of the knee condition. The role of physical therapy and cooperation between the physical therapist and the surgeon is of paramount importance for the final outcome. More work is needed to correlate the relation of the anatomy of the fat pad to knee symptoms and function. PMID- 8718718 TI - Geometrical relationships between bone plug configuration and patellar tendon graft width. AB - There is a lack of information in the orthopedic literature regarding the optimal shape of the patellar and tibial bone plugs when using the central one third of the patellar tendon graft for reconstruction of the anterior cruciate ligament. Clinically, we have observed that the final bone plug and patellar tendon graft width are narrower than the intended width of the graft secondary to contouring the bone plugs to accommodate the bone tunnel. This study mathematically evaluates the relationship between the bone plug cross-sectional configuration and its influence on osseous area and patellar tendon graft width. This mathematical study was divided into three parts using accepted trigonometric and geometric methods. The first part compares the maximum cross-sectional area of four commonly used bone plug shapes: a triangle, rectangle, trapezoid, and square for 8-, 9-, 10-, and 11-mm wide bone-patellar tendon-bone grafts. Area values were calculated as a function of the radius of a circle (bone tunnel) and were compared to determine which configuration provided maximum fill of the tunnel. In the second part, the minimum tunnel diameter needed to accommodate a graft of a given width (8 to 11 mm) for each of the four bone plug shapes was calculated. In Part 3, the maximum bone plug width that would fit in a tunnel of a given diameter (8 to 11 mm) was calculated for each of the four bone plug shapes. Results from Part 1 showed a 47% increase in the area of each shape from 8- to 11 mm wide grafts. Regardless of the bone plug size or tunnel diameter, a square plug filled 64% of the area of a circle; a trapezoid, 59%; a rectangle, 51%; and a triangle, 41%. In Part 2, the minimum tunnel diameter required to accommodate a square was 4 mm greater than the desired graft width for 8- and 9-mm wide grafts, and 5 mm greater for 10- and 11-mm wide grafts. The minimum tunnel diameter needed to accommodate a trapezoidal, rectangular, or triangular-shaped bone plug was only 2 mm greater than the desired graft width. In Part 3, the maximum graft width of the trapezoid, rectangle, and triangle was 0.8 to 1.5 mm smaller (10% to 14% less) than the corresponding tunnel diameters of 8 to 11 mm. Yet, the maximum width of the square was 2.3 to 3.2 mm smaller (29% less) than the corresponding tunnel diameters of 8 to 11 mm. These results confirm our clinical observation that the actual bone plug that would fit the bone tunnel is much narrower than the intended graft width. It is recommended that a trapezoidal or rectangular bone plug be harvested as these two shapes would provide a satisfactory amount of bone (cross-sectional area) compared with a square or triangle, with less tunnel bone plug size difference. PMID- 8718719 TI - Total knee arthroplasty. What have we learned? PMID- 8718720 TI - Prosthetic selection in total knee arthroplasty. PMID- 8718721 TI - Surgical techniques and the management of fixed deformities in total knee arthroplasty. PMID- 8718722 TI - Revision total knee arthroplasty due to aseptic failure. AB - Revision TKA for aseptic failure presents the surgeon with a variety of technical problems. The major problems include loss of bone stock and inadequacy of soft tissue constraints. Historically, these patients have done poorly but when these problems are adequately addressed with the appropriate surgical technique and appropriate choice of implant, then the results are quite good. PMID- 8718723 TI - Diagnosis and management of the infected total knee arthroplasty. AB - While infection in TKA is a relatively infrequent complication, it can be devastating in terms of morbidity and cost. Prevention of infection begins with patient selection. Prior knee sepsis surgery, rheumatoid arthritis, and poor general health may lead to an increased rate of infection. Prophylactic antibiotics, meticulous surgical technique, and control of the intraoperative environment have been shown to be beneficial in prevention of infection after TKA. Diagnosis can be difficult and often is heralded by the onset of pain in a previously pain-free knee. Aspiration is an excellent screening tool and is also beneficial in determining management of potentially infected TKAs. In cases posing a diagnostic dilemma, radiographs and nuclear medicine studies also may prove beneficial as well as intraoperative frozen section. Management is based on chronicity of the infection and fixation of the components. Antibiotic suppression is unlikely to yield a cure but may be indicated in the medically infirm. Debridement with component retention may be used with varying degrees of success, especially in the acute postoperative period. The current treatment of choice for chronic infections in this country is a two-stage revision with interim intravenous antibiotics. This would be expected to yield a cure in approximately 80% of patients. Arthrodesis may be necessary in the patient who is status post-multiple revisions with particular virulent organisms. Resection arthroplasty should be reserved for the older rheumatoid patient with limited functional demands. Finally, amputation should be considered in the patient with life-threatening sepsis or the patient who is status post-multiple revisions with intractable pain and poor bone stock. PMID- 8718724 TI - Standards set internationally for informed consent in health care. PMID- 8718725 TI - Secrets of the couch and the grave: the Anne Sexton case. PMID- 8718726 TI - Must physicians reveal their wounds? PMID- 8718727 TI - Informed consent revisited. PMID- 8718728 TI - How important is consent for controlled clinical trials? PMID- 8718729 TI - Determinants of decision making for circumcision. PMID- 8718730 TI - Informed consent issues in international research concerns. PMID- 8718731 TI - Consent & confidentiality: the shifting paradigm. PMID- 8718732 TI - Is the treatment beneficial, experimental, or futile? PMID- 8718733 TI - Exploring the role of religion in medical ethics. PMID- 8718734 TI - Measurement of moral development in medicine. PMID- 8718735 TI - From a Dutch perspective: response to "Rights of the Terminally Ill Act of the Australian Northern Territory" by Robert L. Schwartz (CQ vol 5, no 1) PMID- 8718736 TI - Organs for undocumented aliens--another look: response to "Distributing American hearts for transplantation: the predicament of living in the global village" by Henry S. Perkins (CQ vol 4, no 2) PMID- 8718737 TI - Task force on standards for ethics consultation: response to "Ethics consultation: the least dangerous profession?" (CQ vol 2, no 4) PMID- 8718738 TI - Reflections on non-heartbeating organ donation: how 3 years of experience affected the University of Pittsburgh's Ethics Committee's actions. PMID- 8718739 TI - Orphan tests. PMID- 8718740 TI - The Canadian question: what's so great about intelligence? PMID- 8718741 TI - Evaluation of commercially available ELISA test kits for the detection of clenbuterol and other beta 2-agonists. AB - This article presents the results of a comparative study of nine commercially available ELISA test kits for clenbuterol and other beta 2-agonists currently being used in veterinary drug residue control of live and slaughtered animals in the European Union (EU). By determining measuring ranges, B/B0-50% values, inter- and intra-assay variations and cross-reactions as well as limits of detection (LOD) and recoveries for clenbuterol in bovine urine it could be demonstrated that a number of test kits showed considerable quality defects, reducing their applicability to residue control. LODs for urine assayed without a previous clean up (as recommended by all kit manufacturers) varied between 1.2 and 11.1 ng clenbuterol per ml urine and did thus not meet the requirement of 1 ng/ml of the official residue control plans of the EU Member States. PMID- 8718742 TI - The effect of cooking on veterinary drug residues in food: 4. Oxytetracycline. AB - The heat stability of oxytetracycline (OTC) in water and vegetable oil was investigated. Results showed that the drug was unstable in water at 100 degrees C with a half-life of about 2 min, but more stable in oil at 180 degrees C where the half-life was about 8 min. The effect of a range of cooking processes including microwaving, boiling, roasting, grilling, braising and frying on OTC residues in incurred animal tissues was investigated. Substantial net reductions in OTC of 35-94% were observed, with temperature during cooking having the largest impact on the loss. Migration from the tissue into the surrounding liquid or meat juices was observed during the cooking processes. Diode-array analysis of heat-treated OTC standard solutions indicated that no individual closely related compound such as 4-epioxytetracycline, alpha- or beta-apooxytetracycline formed a significant proportion of the breakdown products. OTC was not evenly distributed throughout the tissue, but the effects of this were minimized by selecting adjacent samples for cooking and for the raw control. The findings of this investigation showed that the effect of cooking on residues of OTC should be considered before data obtained from measurements on raw tissue are used for consumer exposure estimates and dietary intake calculations. PMID- 8718744 TI - Theoretical treatment of pollutant transfer in a finite volume of food from a polymer packaging made of a recycled film and a functional barrier. AB - The problem of pollutant transfer in food from a polymer packaging made of a recycled film and a functional barrier is studied from a theoretical point of view. In the present case, the process is controlled by Fickian diffusion through the two films of the polymer packaging in perfect contact and by convection into the food. A numerical model is built, taking all the facts into account: the diffusion of the pollutant through the two polymer films, a convective coefficient of mass transfer on the surface, a partition factor, the volume of the liquid and the thicknesses of the polymer films. The results are expressed by using dimensionless numbers so that they can be used for various cases, viz the kinetics of pollutant transferred into the food and the profiles of concentration of the pollutant through the packaging. PMID- 8718743 TI - Residue depletion in tissues and fluids from swine fed sulfamethazine, chlortetracycline and penicillin G in combination. AB - Twenty-four hogs were fed a ration for 14 days containing three times the recommended label dose of a combination drug which included sulfamethazine, chlortetracycline and penicillin G. Groups of six hogs were slaughtered 0, 2, 4, or 8 days after withdrawal. Six untreated control hogs were slaughtered 5 days before the first group, of six treated hogs, were slaughtered. Residue concentrations were determined in kidney, liver, muscle, serum and urine. At zero withdrawal the kidney from one hog contained 0.018 mg penicillin G per kg and the serum from the same hog contained 0.016 mg penicillin G per litre. Penicillin G was not detected in any other samples that were analysed. Chlortetracycline concentrations in tissues at zero withdrawal time were below accepted Canadian Maximum Residue Limits (MRL) for chlortetracycline of 1 mg/kg in muscle, 2 mg/kg in liver and 4 mg/kg in kidney and were below the limit of quantitation in all tissues 4 days after withdrawal. Sulfamethazine persisted in the tissues longer than penicillin G or chlortetracycline. Sulfamethazine concentrations were above the Canadian MRL of 0.1 mg/kg at zero withdrawal time and did not decrease to below the MRL until 8 days after withdrawal. Our results suggest that, if the label withdrawal period of 10 days is observed, an increase in the dosage of up to three times the recommended rate is unlikely to increase significantly the risk that residues would occur in the tissues of treated hogs at concentrations which exceed MRLs. Sulfamethazine concentrations in all matrices decreased after storage at -76 degrees C for 6 months. PMID- 8718745 TI - Oligomers in plastics packaging. Part 1: Migration tests for vinyl chloride tetramer. AB - Vinyl chloride (VC) tetramer has been studied as a representative oligomer that has the potential for migration from plastics packaging. Poly(vinyl chloride) (PVC) bottles for retail beverages were analysed by a process of dissolution followed by gas chromatography. Tetramer levels ranged from 70 to 190 mg/kg in the plastic. When these bottles were tested for migration into the simulants distilled water, 3% acetic acid, 15% ethanol and olive oil, no tetramer migration was detected at a limit of 5-10 micrograms/kg. Since, of the low molecular weight oligomers, the tetramer had the highest concentration in the PVC plastics, it is concluded that the other VC oligomers of higher molecular weight, would not migrate above this limit of detection either. PMID- 8718746 TI - Effect of dichlorvos on growth and mycotoxin production by Alternaria alternata. AB - The effect of dichlorvos (DDPV) on growth and alternariol (AOH), alternariol monomethyl ether (AME) and tenuazonic acid (TA) production was evaluated both in culture media and on sunflower seeds. At the levels evaluated of 1 to 600 mg/kg, the insecticide showed a slight effect on the linear growth rate, which was dose dependent, and caused morphological changes in the mycelia and conidia. The pesticide had a remarkable influence on AOH, AME and TA production. PMID- 8718747 TI - Occurrence of the mycotoxin moniliformin in maize (Zea mays L.) ears infected by Fusarium subglutinans (Wollenw. & Reinking) Nelson et al. AB - Fusarium subglutinans has been identified as a prevailing pathogen of maize ears in Poland in the seasons 1985-1991. About 95-100% of ears with Fusarium ear rot symptoms were infected by this species. Moniliformin was present in all 57 ears with pink ear rot symptoms examined. Fusarium-damaged kernels contained an average of 130.9 mg/kg of moniliformin, with large differences between individual samples each year--from 4.2 mg/kg to 530 mg/kg. PMID- 8718748 TI - Aflatoxin B1 contamination in groundnut samples collected from different geographical regions of India: a multicentre study. AB - Under a multicentre study conducted by the Indian Council of Medical Research, 2062 samples of groundnut were collected from rural and urban areas of 11 states representing different geographical regions of the country. These samples were analysed for aflatoxin B1 using the AOAC method. Analytical quality assurance between various participating laboratories was ensured through analysis of check samples. Twenty-one per cent of groundnut samples collected from 11 states exceeded the permissible Indian regulatory limit of 30 micrograms/kg. No statistically significant difference in percentage of samples contaminated (> 30 micrograms/kg) were observed between pooled rural (22.9%) and urban (19.9%) data. Amongst the 11 states, the minimum and maximum per cent contamination respectively (> 30 micrograms/kg) was observed to be 15.2 in the state of Andhra Pradesh and 28.3 in the state of Haryana. The maximum level of contamination of 833 micrograms/kg was observed in the state of Gujarat. The median level of < 5 micrograms/kg was observed in the states of Gujarat, Haryana, Punjab, Andhra Pradesh and Karnataka. In all other states studied, the median level was found to vary between 10 and 20 micrograms/kg. The 90th percentile values were high in Andhra Pradesh (125 micrograms/kg), Gujarat (111 micrograms/kg) and Haryana (110 micrograms/kg). In the remaining states the 90th percentile value ranged from 60 to 93 micrograms/kg. Analysis of pooled data showed the percentages of samples exceeding the level of contamination of 5 micrograms/kg and 15 micrograms/kg, respectively were 44.9% and 37.4% which therefore, showed a marked increase when compared with the per cent of samples exceeding 30 micrograms/kg in the overall data. PMID- 8718749 TI - Survey of natural occurrence of trichothecene mycotoxins and zearalenone in Korean cereals harvested in 1992 using gas chromatography/mass spectrometry. AB - For the survey of the natural occurrence of trichothecene mycotoxins, produced by species of fungi imperfecti such as Fusarium and Trichothecium, a sensitive analytical method was developed for the simultaneous detection and quantitation of the major trichothecene mycotoxins, viz. T-2 toxin (T-2), HT-2 toxin (HT-2), nivalenol (NIV), fusarenon-X (F-X), deoxynivalenol (DON), 3-acetyl deoxynivalenol (3-Ac DON), and zearalenone (ZEN), using gas chromatography/mass spectrometry selected ion monitoring (GC/MS-SIM) mode after trimethyl silyl derivatization. The incidence of NIV and DON in 30 barley samples were 93% and 67%, respectively; the average contents of NIV and DON in positive samples were 390 ng/g (range 40 2038) and 106 ng/g (range 5-361) respectively. In 15 maize samples, the incidences of NIV and DON were 53% and 93% respectively and the average contents were 168 ng/g and 145 ng/g, respectively. These results suggest that NIV and DON were the major contaminating trichothecene mycotoxins in Korean barley and maize samples harvested in 1992. PMID- 8718750 TI - Analytical methods for the determination of sterigmatocystin in cheese, bread and corn products using HPLC with atmospheric pressure ionization mass spectrometric detection. AB - Methods have been developed for the determination of sterigmatocystin in bread, maize and cheese using HPLC linked to mass spectrometry (MS) atmospheric pressure chemical ionization for detection and concurrent confirmation of sterigmatocystin at levels down to less than 5 micrograms/kg. Candidate extraction methods were initially checked for recovery and reproducibility by spiking commodities at a level of 200 micrograms/kg of sterigmatocystin and using HPLC with post-column derivatization. Recovery was found to be greater than 90% for bread and maize, and 75% for cheese. Mass spectrometer conditions for detecting sterigmatocystin were established by injecting solutions directly into the mass spectrometer. Extracts of bread, maize grits and cheese prepared by the candidate extraction methods were examined using HPLC/MS using samples spiked at a level of 20 micrograms/kg of sterigmatocystin. Results for bread and maize samples showed that the extraction procedure recovered more than 90% of added sterigmatocystin and produced extracts free of interference from co-extractives, with limits of detection of less than 2 micrograms/kg for both commodities. The HPLC/MS results for cheese extracts gave lower average recoveries of 55%. These results were also more variable. However, the apparent limit of detection for sterigmatocystin in cheese was still about 4 micrograms/kg. PMID- 8718752 TI - Salable goods. PMID- 8718751 TI - Oral cadmium exposure of adults in Germany. 1: Cadmium content of foodstuffs and beverages. AB - The cadmium contents of 94 and 105 foodstuffs bought in six-fold repetition in 1988 and in nine-fold repetition in 1991, respectively were analysed within the framework of a market-basket study. These foodstuffs were typical of German eating habits. Additionally, 170 samples of drinking water were investigated. The cadmium concentrations of the foodstuffs were comparable with results of recent studies carried out in Europe and North America. Fruit, milk and dairy products, sugar and sugar-rich foodstuffs as well as beverages showed mean cadmium contents < or = 5 ng/g fresh matter or ng/ml, respectively. The cadmium content of meat, sausage, fish and tinned fish was also low. Pork and beef, the most important kinds of meat, contained 5.4 and 2.5 ng/g on average. The majority of the vegetables investigated, including potatoes, had cadmium concentrations < 25 ng/g. However, individuals samples of lettuce showed very high cadmium levels. The cadmium content of bread, cakes and pastries as well as farinaceous products were within the range of 20-40 ng/g. The most important bread, cakes and pastries (wheat and rye bread, toasted bread, rolls) contained 25-35 ng/g. A median cadmium concentration of 0.2 micrograms/l was found in the drinking water. As expected, liver and kidneys showed the highest cadmium levels of 73 and 204 ng/g, respectively on average. PMID- 8718753 TI - An open labelled assessment of adverse effects associated with interferon 1-beta in the treatment of multiple sclerosis. PMID- 8718754 TI - Anatomic correlates of psychological events related to temporolimbic epilepsy. AB - A number of behavioral disturbances have been associated with epilepsy. This is particularly true of temporal lobe seizures. While many ictal behaviors such as automatisms and rhythmic movement of an extremity are recognized as seizure activity, many psychological and cognitive alterations are not appreciated as being related to epilepsy. An increased awareness of the range of possible behaviors associated with epilepsy will enhance the observations made by the neuroscience nurse and augment the information available for formulating a differential diagnosis and localizing the lesion. Appropriate explanation of the etiology and manifestation of temporal lobe seizures will also help educate the patient and family regarding the unusual behaviors which are part of this disorder and reassure them of the organic rather than psychiatric cause of the phenomena. PMID- 8718755 TI - A model of family adaptation to new-onset childhood epilepsy. AB - The onset of a chronic health condition such as epilepsy can interfere with the successful accomplishment of important psychosocial developmental tasks. Children with epilepsy are especially at risk for problems and often exhibit poor self esteem, social withdrawal, and behavior problems. The purposes of this paper are first, to review the relevant literature on factors related to psychosocial adaptation in childhood epilepsy, and second, to present a model based on this literature to guide psychosocial nursing practice. Implications for psychosocial nursing care are discussed. Moreover, instruments for use in assessment of children and families are described, and instruments to assess need for psychosocial nursing care are presented. PMID- 8718756 TI - Controversies in the management of carotid stenosis. AB - Controversies surround the indications for and methods of diagnosis of carotid stenosis. Carotid endarterectomy is of proven value for patients presenting with transient ischemic attacks or minor stroke and angiographic atherosclerotic carotid stenosis of greater than 70% if endarterectomy can be carried out with a surgical morbidity and mortality of less than 6%. It is generally not indicated for stenosis less than 30%. For intermediate levels of stenosis, studies to determine its efficacy are ongoing. Endarterectomy can also decrease the risk of stroke in asymptomatic patients, although indications in asymptomatic patients remain controversial. Conventional angiography remains the gold standard for diagnosis of carotid stenosis. Improvements in ultrasound technology and magnetic resonance angiography have led some surgeons to rely only on these tests preoperatively rather than perform conventional angiography. It would be advantageous to be able to avoid the 1% risk of angiography but further study is indicated before this approach can be accepted. Carotid angioplasty remains an investigational procedure. PMID- 8718757 TI - Carotid endarterectomy: a nursing perspective. PMID- 8718758 TI - Sexuality and related issues in the preadolescent and adolescent female with epilepsy. AB - Multiple resources for the adult female with epilepsy are noted in the literature. Topics covered include pregnancy, epilepsy, contraception, anticonvulsant medications and issues of sexuality. Adolescence is a time of physiological and psychosocial change. The adolescent faces the usual challenges of this life stage along with the added stress of issues surrounding epilepsy. There is minimal information available for and about adolescents with epilepsy. PMID- 8718759 TI - Innovations in aneurysmal subarachnoid hemorrhage: intracisternal t-PA for the prevention of vasospasm. AB - Aneurysmal subarachnoid hemorrhage (SAH) affects approximately 30,000 people each year in North America. At least 30% of these patients will develop vasospasm as a result of the initial hemorrhage, and two thirds of these develop permanent disabilities or die. Blood deposited into the basal cisterns from the ruptured aneurysm can form thick clots around the major cerebral vessels. The by-products of the hemolyzed clots are believed to be responsible for the subsequent development of vasospasm. Hypervolemic, hypertensive, hemodilution therapy (HHHT) and nimodipine may improve outcome in some cases but there is no therapy known to prevent vasospasm in all patients. One potential therapeutic agent under investigation is tissue plasminogen activator (t-PA), a fibrinolytic enzyme. Instilled into the basal cisterns at time of aneurysm clipping, t-PA dissolves the clot so spasmogenic substances may be removed, thus preventing or reducing the severity of vasospasm. PMID- 8718760 TI - The role of interferons in the treatment of multiple sclerosis. AB - IFN beta-1b clearly alters the course of disease in patients with relapsing remitting MS, although the adverse effects of this agent may discourage some patients from continuing the therapy. Nurses can help patients with MS minimize adverse effects through teaching. A double-blind, placebo-controlled clinical trial in patients with relapsing-progressive MS will be started in 1995. It is hoped that IFN beta-1b will be effective in slowing the accumulation of CNS lesions in those with progressive MS. PMID- 8718761 TI - Feminist research. PMID- 8718762 TI - Priming, analogy, and awareness in complex reasoning. AB - The mechanisms by which a concept used in solving one complex task can influence performance on another complex task were investigated. We tested the hypothesis that even when subjects do not spontaneously make an analogy between two domains, knowledge of one domain can still spontaneously influence reasoning about the other domain via the mechanism of priming. Four groups of subjects (two experimental and two control) were given a simulated biochemistry problem on Day 1 and a simulated molecular genetics problem on Day 2. For the two experimental groups, the solution to the biochemistry problem involved inhibition. For the two control groups, the solution did not involve inhibition. On Day 2, all subjects received the same version of the molecular genetics problem in which the solution involved the concept of inhibition. Subjects in the experimental conditions were more likely to attain the correct answer, to propose inhibition, and to propose inhibition early in the problem-solving session than were subjects in the control conditions. However, subjects in the experimental conditions made no reference to the biochemistry problem either in their verbal protocols or in a post-task questionnaire. The results are interpreted as demonstrating that an implicit process--priming--can make old knowledge available for current problem solving. PMID- 8718763 TI - The importance of being upright: use of environmental and viewer-centered reference frames in shape discriminations of novel three-dimensional objects. AB - We investigated the frame of reference that people use to make shape discriminations when their heads are either upright or tilted. Observers made same-different judgments of pairs of novel three-dimensional objects that were aligned along their length within the frontal-parallel plane and rotated in depth around an axis parallel to their own axes of elongation. The aligned objects were displayed vertically, tilted 45 degrees, or horizontally with respect to the environmental upright, but the distance of each pair from the upright was irrelevant to resolving the angular disparity between the stimuli for the same different judgment. Nevertheless, when observers' heads were upright, the time to encode the stimuli was a linear function of the distance of the stimuli from the environmental upright, whereas when observers' heads were tilted 45 degrees, encoding times for tilted and vertical stimuli did not differ and were faster than the times to encode horizontal stimuli. We interpreted these data to mean that observers either rotate or reference the top of an object to the environmental upright, and they can use either a gravitational or retinal reference frame to do so when either they or the objects are not upright. PMID- 8718764 TI - From auditory image to auditory percept: facilitation through common processes? AB - Two experiments explored implicit memory for auditory stimuli as measured by a test of perceptual identification. The facilitative effect of perceived auditory primes was contrasted with that of imaged auditory primes. In Experiment 1, there was a significant priming effect from imaged spoken-word primes that did not differ significantly from the level of priming due to perceived spoken-word primes, measured by a test of auditory perceptual identification. There was no facilitation of spoken-word identification following creation of an image of a word's referent sound. In Experiment 2, identification of an environmental sound was facilitated by prior processing of an imaged sound from the same category, though there was significantly more transfer following processing of the actual sound. PMID- 8718765 TI - Group aggregates and individual reliability: the case of verbal short-term memory. AB - Two experiments examined the generalizability of the effects of word length and phonological similarity with visual and auditory presentation in immediate verbal serial ordered recall. In Experiment 1, data were collected from 251 adult volunteers drawn from a broad cross-section of the normal population. Word length and phonological similarity in both presentation modes significantly influenced the group means. However, 43% of the subjects failed to show at least one of the effects, and the likelihood that effects appeared was highly correlated with verbal memory span. In Experiment 2, 40 subjects of the original sample were retested, 20 of whom had failed to show one or more effects in Experiment 1. Whether or not an effect had appeared for individual subjects on the first test session was a poor predictor of whether the effect would appear on retest. Finally, an analysis of subject reports demonstrated that the patterns of experimental data could be accounted for in part by the strategies that subjects reported using, and the effect of strategy was independent of the effect of span. The implications of these findings for theories of verbal short-term memory are discussed. PMID- 8718766 TI - The representation of color and form in long-term memory. AB - Color and form are elementary stimulus encoding dimensions that have effects on the representation of visual stimuli at early processing stages. Little is known, however, about their effects on visual long-term memory. In three experiments we investigated whether color is part of the memory representation, whether color and form are bound in the memory representation, and the effect of color context on memory performance. Experimental results suggest that color is part of the memory representation and that color and form can be represented separately in memory and accessed independently. We suggest that the binding of color and form is a deliberate strategic act that requires focal attention, not a natural consequence of processing visual stimuli. We compare our results with the predictions of two computational memory models regarding feature binding. The effect of color context was not straightforward; however, results are consistent with the encoding specificity principle. PMID- 8718767 TI - The word-detection effect: sophisticated guessing or perceptual enhancement? AB - Subjects discriminate letters in words better than letters in nonwords. The sophisticated guessing hypothesis attributes this word advantage to a guessing strategy. In words, the possible letters at each letter position are constrained by letters at other positions, whereas letters in nonwords are not restricted in this manner. A critical test of this hypothesis is that if subjects are given explicit knowledge of the letters in nonwords before the trial, the word advantage would disappear. We investigated the effect of preknowledge of the alternatives in the word-detection effect. In the word-detection effect, subjects decide which of two character strings contains letters and which contains pseudoletters. In four experiments, subjects were more accurate with words than with nonwords, and subjects were more accurate when they were told the word or nonword before the trial. However, even with foreknowledge of the alternatives, subjects were more accurate with words than with nonwords. PMID- 8718768 TI - Processing resource capacity and the comprehension of garden path sentences. AB - Three experiments explored the relationship between verbal working memory capacity and the comprehension of garden path sentences. In Experiment 1, subjects with high, medium, and low working memory spans made acceptability judgments about garden path and control sentences under whole sentence and rapid serial visual presentation (RSVP) conditions. There were no significant differences between subjects with different working memory spans in the comprehension of garden path sentences in either condition. In Experiments 2A and 2B, subjects with high and low working memory spans were tested on the same materials at three RSVP rates. There were no significant differences between subjects with different working memory spans in the magnitude of the effect of garden path sentences at any presentation rate. The results suggest that working memory capacity, as measured by the Daneman and Carpenter (1980) reading span task, is not a major determinant of individual differences in the processing of garden path sentences. PMID- 8718769 TI - The relation between discriminability and memory for vowels, consonants, and silent-center vowels. AB - People remember lists of vowel-contrasting syllables better than lists that vary only in stop consonant identity. Most views suggest that this difference is due to the structure of immediate memory and the greater discriminability of vowels compared with consonants. In all of these views, there is a presumed systematic relationship between discriminability and recall so that the more discriminable an item, the better that item should be recalled. The 11 experiments reported here measured the relative discriminability of and compared serial recall for (1) intact syllables that varied only in the medial vowel, (2) intact syllables that varied only in the initial consonant, and (3) syllables with the center vowel replaced by silence (so-called silent-center vowels). When item discriminability, as measured by identification, was equated for consonant-contrasting and silent center lists, serial recall performance was also equal. However, even when the vowels were less discriminable than the consonants or silent-center vowels, serial recall performance for the vowels was still better. These results are problematic for theories based on acoustic discriminability but can be explained parsimoniously by Nairne's (1990) feature model. PMID- 8718770 TI - The sensory match effect in recognition memory: perceptual fluency or episodic trace? AB - The sensory match effect in recognition memory refers to the finding that recognition is better when the sensory form in which an item is tested is the same as that in which it was studied. This paper examines the basis for the sensory match effect by manipulating whether a studied fragmented picture is tested with the same or a complementary set of fragments in a recognition memory test (Experiment 1) and in a fragment-identification test (Experiment 2). Assuming that fragment identification is a direct measure of perceptual fluency, we expected identical patterns of results across the two tests if perceptual fluency accounted for the sensory match effect in recognition memory. Instead, recognition memory showed a robust overall sensory match effect (the same fragmented image was recognized better than the complementary image), whereas fragment identification showed no overall sensory match effect (the same fragmented image was identified no better than the complementary fragmented image). Experiments 3 and 4 combined the two responses and showed that the basis for the sensory match effect in recognition memory was a subject's ability to recognize the matching fragments in the absence of conceptual information (when the test stimulus could not be identified), supporting the idea that the episodic trace of the sensory code is responsible for the sensory match effect in recognition memory. Experiment 5 demonstrated that subjects are able to use this sensory code as the sole basis for recognition memory. PMID- 8718771 TI - The influence of medical expertise, case typicality, and illness script component on case processing and disease probability estimates. AB - The present study investigated the influence of medical expertise, case typicality, and illness script component (enabling conditions vs. consequences) on the speed of case information processing and subjective disease probabilities. It was hypothesized that expert subjects would process case information faster than nonexpert subjects, that typical information would be processed faster than atypical information, and that an interaction would be found between expertise level, typicality, and illness script: Experts were expected to be sensitive to typicality of both illness script components, while advanced students would be sensitive only to typicality of consequences. This sensitivity would also be reflected in assigned probability estimates. The data supported the predictions concerning the effects of expertise level and typicality; it was also found that expert physicians are particularly sensitive to a combination of prototypical enabling conditions and prototypical consequences. Implications of these results for the illness script theory are discussed. PMID- 8718772 TI - Undermining prejudice: the other epidemic. PMID- 8718773 TI - Drinking problems of nursing students. AB - A sample of 315 nursing students in yearly cohorts (1988-1992) completed the Michigan Alcoholism Screening Test (MAST). A simple and quick instrument with established reliability and validity, the MAST had a reliability coefficient of .78 with this sample. The MAST scores of the nursing students indicated that 21.5% had probable alcoholism or were alcoholic. Drinking problems among these nursing students were similar to those of other nursing students and college students in general. Implications for nurse educators include recognition that nursing students need continuing self-assessment and monitoring to avoid drinking problems as a student and future practitioner. Alcoholism self-screening is an excellent introduction to client assessment for drinking problems. Implications for future research include identifying correlates and determinants of problem drinking behaviors and ultimately developing and testing educational interventions to eliminate the problem. PMID- 8718774 TI - Heterosexism in nursing education. AB - If we are to take seriously the challenge of teaching nursing students skills in critical thinking, as nurse educators we must examine the most difficult issues facing society and nurses. Heterosexism and the resulting homophobia are such issues. This article provides an introduction to the concepts of heterosexism and homophobia and describes how they intersect and are revealed in nursing education. The consequences of heterosexism and homophobia are illustrated using examples from our experiences as lesbian and heterosexual nurse educators. We conclude with an action plan for undermining heterosexism and homophobia. PMID- 8718775 TI - The impact of violence in the health care setting upon nursing education. AB - As the incidence of violence and potential risks for violence have increased in the health care workplace, the safety of nurses and nursing students has emerged as a critical concern for the profession. This article provides a review of the literature focusing on the incidence of violence toward nurses, factors that contribute to the occurrence of violence, the management of violent or potentially violent situations, and the effects on nurse victims of violence. Additionally, the costs of violence to the health care system and the various ways that violence has been defined are reviewed. The literature on violence occurs primarily in the clinical specialty areas of psychiatric-mental health, emergency department, long-term care, and home care nursing. To address the need to prepare students to deal with the issue of violence in the health care setting, strategies for nursing education are presented which include role playing, videotape playbacks, and debriefing sessions. Suggested content consists of student self-awareness, assessment, and diagnosis of violent or potentially violent clients, and nursing care planning, interventions, and evaluation for the immediate situation, as well as long-term treatment goals. The authors emphasize the obligation of nurse educators to prepare students to deal with violence in the health care setting. PMID- 8718776 TI - Inquiry based learning, nursing student attitudes and the HIV patient. PMID- 8718777 TI - A critical experiential teaching strategy: student and faculty participation in an AIDS walk. PMID- 8718778 TI - Intervention to promote safer sexual behaviors and educate peers about HIV/AIDS. AB - Sexually active college students are at risk for infection with the Human Immunodeficiency Virus (HIV). Nursing students can learn a variety of desirable professional skills and provide an important service to their college peers by participating in an ongoing intervention to promote safer sexual behaviors. Since 1989, nursing students at a publicly supported urban college have been providing a HIV Primary Prevention intervention on a sustained basis. During each semester, at least 1,000 college students complete a quiz that tests their knowledge about condoms and engage in discourse with a nursing student about HIV/AIDS. Content analysis of the evaluations submitted by the nursing students (N = 43) identifies how this independent study enhances both professional and personal growth. PMID- 8718779 TI - HIV/AIDS nursing care: instrument for measuring students' attitudes and precautions. PMID- 8718780 TI - Stigma: baccalaureate nursing students' attitudes toward persons living with AIDS. PMID- 8718781 TI - Attitudes and perceptions of nursing students toward chemically impaired nurses: implications for nursing education. PMID- 8718782 TI - Analysis of receptors for cytokines and growth factors in human disease. AB - The physiological importance of cytokines and other factors which control cell and tissue growth and differentiation is widely appreciated. While physiological studies have included the cellular receptors for these factors, studies on their role in disease have concentrated on the measurement of cytokines themselves or soluble receptor components. This reflects in part the technical difficulty of measuring cell surface expression of receptors, which occur at and are functional at very low concentrations. In this review, the potential value of surface expressed receptors as markers of disease is assessed, and methods are described which allow measurements with standard equipment for flow cytometry and fluorescence microscopy. PMID- 8718783 TI - Establishment of sequence-tagged sites on 15q11-q13 by Alu-vector PCR cloning of YAC-generated fragments. AB - Angelman syndrome (AS) is caused by the loss of function of undefined gene(s) on human chromosome 15. The majority of subjects have deletions involving maternally derived chromosome 15q11-q13, and the shortest region of deletion overlap (SRO) has been localized to the region between D15S10 and D15S113. In this study, yeast artificial chromosomes (YACs), 6G-D4, 9H-D2 and 37D-F9, mapping within the AS SRO, were isolated from the ICI YAC library. Alu-vector PCR products were amplified from the YACs and from YACs A229A2 and A33F10 which had been obtained from the St. Louis YAC library. The PCR products were cloned and sequenced, and three new sequence-tagged sites were generated within the AS SRO, facilitating the characterization of gene(s) involved in the Angelman syndrome. PMID- 8718784 TI - HLA-B27 determination by polymerase chain reaction. AB - A method for determining the presence or absence of HLA-B27 by selective amplification of a region in the third exon of the HLA-B27 gene common to B*2701 to B*2705 inclusive, was evaluated. This polymerase chain reaction/sequence specific primer (PCR-SSP) method gave perfect correlation with serological typing on 40 individuals of previously determined HLA type and on 50 further clinical samples evaluated blind. It was concluded that HLA-B27 determination by PCR-SSP is simple, reliable, cost-effective and convenient for laboratory staff. PMID- 8718785 TI - Cathepsin-D and TNF-alpha in bladder cancer. AB - In a study of 34 normal healthy controls, 35 patients with urinary tract bilharziasis and 93 bladder cancer patients (62 of them are operable cases and 31 are non-operable ones), serum tumor necrosis factor alpha (TNF-alpha) and cytosolic Cathepsin-D were estimated. Though both potential markers were elevated in bladder cancer patients, neither Cathepsin-D nor TNF-alpha showed associations of prognostic value since there were no positive correlations with tumor stages, grades or association of tumors with bilharzial ova or lymph node involvement. PMID- 8718787 TI - Biotechnology information in Europe: a synopsis of the final report from the Biotechnology Information Strategic Forum. PMID- 8718786 TI - Phosphotyrosine-protein-phosphatases and human reproduction: an association between low molecular weight acid phosphatase (ACP1) and spontaneous abortion. AB - ACP1 (low molecular weight acid phosphatase) genetic polymorphism has been studied in 173 women with a history of two or more consecutive spontaneous abortions and in 1508 control subjects, including 482 normal pregnant women. The proportion of carriers of ACP1*C allele (*A/ *C, *B/*C) in women with a history of repeated spontaneous abortion is lower than in normal pregnant women and other control groups. Women with repeated spontaneous abortion show a specific decrease of ACP1 S isoform concentration as compared to normal pregnant women. The other component of ACP1 activity, the F isoform, does not show a significant difference between the two groups. The data suggest that women with ACP1 genotypes showing a high concentration of S isoform are relatively 'protected' against spontaneous abortion. Preliminary analysis of a sample of 352 normal puerperae along with their newborn babies supports this hypothesis. PMID- 8718788 TI - Isolation of a cDNA-derived expressed-sequence-tag clone on chromosome 15. PMID- 8718789 TI - IgG avidity in the diagnosis of acute Ross River virus infection. PMID- 8718790 TI - Micronucleoli in lymphocytes of the peripheral blood of patients suffering from chronic lymphocytic leukemia (B type). AB - The incidence of micronucleoli was investigated in lymphocytes of the peripheral blood of patients suffering from B chronic lymphocytic leukemia (B CLL). The cytostatic monochemotherapy with chlorambucil apparently produced in these cells an increased incidence of micronucleoli which are known to represent the morphological expression of the inhibition of the nucleolar ribonucleic acid (RNA) transcription. Similarly, the percentage of lymphocytes with micronucleoli was increased after the cytostatic therapy disregarding whether such nucleoli were present in these cells as the only nucleolar type or in addition to other nucleolar types as satellite nucleoli. PMID- 8718792 TI - [Development and biomechanics of the talocrural joint]. AB - We have studied the development of talocrural joint in fetuses, newborns and children. The results of the microscopical, anatomical and biomechanical studies are as follows: 1. The distal extremities of tibia and fibula are joint not only by the distal part of the tibiofibular syndesmosis, but also by an articulation (articulation tibiofibularis distalis). Its articular cavity communicates with the cavity of the talocrural joint and is visible on its arthrographic pictures. This articulation enables the mutual shift and rotation between distal extremities of tibia and fibula-during the plantar flexion fibula turns and shifts round tibia anteriorly, by the dorsal flexion posteriorly. 2. We recommend to preserve the possibility of mutual movements of tibia and fibula in all cases of the surgical treatment of talocrural injuries. 3. Analysis of prostheses of talocrural joint showed why they cannot be functionally successful. They do not respect the physiological movements, the small articulations of tarsus are overloaded. Pain, arthrosis and functional collapse results in these small articulations, because the prostheses change the axis of movements and the biomechanics at all. 4. We have defined conditions for a successful talocrural prosthesis. 5. Study of the topography of the attachment of the Achilles tendon showed, that it develops medially from the axis of the extremity and therefore rotates in the fetus the foot in the equinovarus position. The tendon shifts till the axis during the second half of fetal life. When this shift is temporarily stopped or slown down, the syndrome of pes equinovarus congenitus develops. We recommend the modification of the surgical treatment of pes equinovarus congenitus: transposition of the Achilles tendon in the axial position. PMID- 8718791 TI - Causes of acute exacerbations of chronic hepatitis B. AB - The authors analyzed acute exacerbations of chronic hepatitis B in 24 chronic HBsAg carriers (20 with positive HBeAg, 4 with anti-HBe), more than half of whom were treated with glucocorticoids, by examining specific antigens and antibodies of the hepatitis A virus (HAV), hepatitis B virus (HBV), hepatitis delta virus (HDV), Epstein-Barr virus (EBV) and cytomegalovirus (CMV). Of 38 observed exacerbations of the disease they found in 7 (18%) deterioration after discontinuation of glucocorticoids, in 7 (18%) seroconversion of HBeAg to anti HBe, in 9 (24%) spontaneous exacerbation of chronic hepatitis B, in 11 (29%) chronic activation, reactivation of EBV superinfection, in 3 (8%) CMV reactivation or superinfection and in one (3%) HDV superinfection. In no instance HAV superinfection was observed. Exacerbations were found five times more frequently in subjects in the stage of active viral replication (in HBeAg+ subjects) than in subjects with anti-HBe. In eight patients EBV reactivation was found after glucocorticoids withdrawal: in five patients after an average period of 5 months (range 3-7) and in three patients after an average period of 33 months (range 22-48). The results provide evidence that in addition to already known causes of acute exacerbations of chronic hepatitis B in our geographical area probably an important cause of deterioration of hepatitis is reactivation or superinfection with EBV and CMV in some patients in conjunction with previous glucocorticoid treatment. PMID- 8718793 TI - [Use of immunohistochemical markers in the diagnosis of bronchogenic tumors]. AB - In an unselected series of 32 biopsies of pulmonary neoplasms comprising 25 with neuroendocrine differentiation (11 carcinoids, 9 small cell cancers, and 5 undifferentiated carcinomas) a panel of antibodies (chromogranin-CG, neuron specific enolasis-NSE, S-100 protein, serotonin, bombesin, cytokeratins AE1-3, CAM5.2, epithelial membrane antigen-EMA) was used. This set was compared with results in another series of 7 non-small cell cancers (4 spinocellular and 3 adenomatous). The results of investigation of both the series were in agreement with the fact that there is no completely satisfactory marker of neuroendocrine differentiation. That is the reason why it is the best to use a spectrum of antibodies even in the routine praxis, as the proof of this differentiation in a neoplasm is most important for the differential diagnosis as well as for the therapy and prognosis. PMID- 8718794 TI - [The effect of physical stress on rheological properties of blood]. AB - The influence of physical load on blood viscosity was observed in the presented study. Three groups of persons were examined: 23 volleyball and football players, 96 employees of a nuclear power plant, and 49 employees of a coal power plant. The values of blood viscosity and hematocrit were determined in the quiescent state before the load, after the performance of 2W.kg-1, and after the maximum performance obtained at a bicycle ergometer. After the data processing the following results were obtained: The average performance of the trained persons was 40% higher compared to the untrained persons (power plants employees). Blood viscosity was lower before the load in the trained persons, and remained on average by 5% lower also after the maximum load. The total relative increase of the value of blood viscosity caused by the maximum load was however approximately the same in all groups (about 15%). The mean value of blood pressure increased with the increased blood viscosity, and therefore also with the increased physical load. The study contains also mathematical expressions of the observed relations. PMID- 8718795 TI - [Thromboembolic complications in urology]. AB - Using the latest examination methods the authors have proved relatively wide spread recurrence of thromboembolic complications. Proved were even the mortal forms in urology in case of benign diagnosis as well as relatively considerate operational medical performance. An evident effect of prophylactic, physical and especially pharmacological methods was proved in prevention of thromboembolic disorders. Silent form of deep venous thrombosis were diagnosed to patients with mere physical prophylaxis. On the other side unfavourable effect of low molecular weight heparin on blood loses was not proved even in laboratory picture. Higher risk of thromboembolic complications origin was observed even on the 21st postoperation day. Currently authors consult the appropriateness of individual types of prophylaxis with the different urology illnesses risk groups. PMID- 8718796 TI - [Motivation for preventive care]. AB - This article outlines those psychological characteristics of preventive and prophylactic work that qualify it as an efficient and effective skill. The transactional analysis model is used for explaining some peculiarities of everyday contact and communication between medical professionals and patients. PMID- 8718797 TI - [Is there an explanation why O. Schrutz, the anatomist, became a medical historian?]. PMID- 8718798 TI - [Prague physicians abroad in the 17th century]. AB - In the early 17th century the Medical Faculty of Prague University was not yet established. Future medical doctors had to study abroad. Because of religious orientation in our country they studied mainly at evangelical universities in Germany. The international relations of Prague students and doctors of medicine became more varied, when the Prague Medical Faculty was established and especially in the second half of the 17th century. At that time many students and doctors from Prague went to study most often to Italy. PMID- 8718799 TI - [Analysis of the literary legacy of Professor Josef Charvat]. AB - The literary inheritance of Professor Joseph Charvat (1897-1984) consists of 10 monographs and 545 other titles, some of which have not appeared in the public press. The author himself arranged his original papers written in Czech as well as other European languages, reviews, essays on health education, publicistic discourses and Varia chronologically from 1922 till 1981. His literary inheritance reflects the history of the Czech internal medicine, the Czech medical faculty of Charles University in Prague and the Czech society and illustrates the broad field of interests of the author. As a disciple of Joseph Pelnar, he was a broadly oriented internist and a classic of the Czech academic medicine. He is recognized as the founder of Czechoslovak endocrinology and one of the founders of behavioural medicine (21). He was interested in medical education (12) and promotion of clinical science and research. He predicted the significance of biocybernetics, clinical genetics and social medicine and public health for the further development of health sciences. His literary inheritance reflects his civic responsibility, courage and authority he enjoyed in the Czech society during the Thirties, the Forties and the Sixties. His work gives evidence of his activities in the World Health Organization and on Economic and Social Council of the Advisory Committee of United Nations, concerning the application of science and technology to the general world development (13, 14, 15, 16). Professor Joseph Charvat's inheritance has not only historical relevance as it contains specific entities, such as holistic approach to medicine, science, men and human society, which should be rediscovered in the present time. Important is also his warning that the progress of humanistic subjects in medicine lags behind the advances in natural science and technology. Professor Charvat offers an interesting concept of creativity. His mastery of communication and his ethical reflexion on scientific knowledge in natural sciences are of lasting value. The dominant principle of his attitude toward problem solving, in medicine and in the society as well, is a positive feedback concept. PMID- 8718800 TI - [Level of maturation of embryonic neurons in suspensions used for neurotransplantation]. PMID- 8718801 TI - [Protective effects of vitamins C and E in hypoxia]. PMID- 8718802 TI - [Do we do research for pleasure even in medicine?]. PMID- 8718803 TI - [The status of Czech science]. PMID- 8718804 TI - Ovarian Cancer. PMID- 8718805 TI - Committee to advise on tropical medicine and travel (CATMAT). Statement on motion sickness. PMID- 8718806 TI - A brief history of the IVth Department of Internal Medicine, 1st Faculty of Medicine, the Charles University and of the General Faculty Hospital in Prague. PMID- 8718807 TI - Bioethical concepts of health in medicine. AB - As a fundamental human need, and consequently one of the principal ethical values, health has to be examined both from the experiential and normative aspects. The following concepts of health have successively developed and found their place in internal medicine: a) absence of manifest disorders, b) state of complete well-being, c) active process aimed at achieving consistency of functions in a dynamic equilibrium with the environment, in order to secure optimum satisfaction of biological and cultural needs. The essential unity of the somatic, psychic, social and spiritual dimensions of the human person should be considered, when assessing health of individual subjects as well as of the whole community. PMID- 8718808 TI - Lyme carditis. PMID- 8718809 TI - The role of intravascular stents in preventing peripheral embolization during pulse-spray thrombolysis. PMID- 8718810 TI - From critical ischaemia to incipient critical ischaemia: a new concept of good hope for salvage of endangered extremities. AB - 1. Although critical limb ischaemia has been clearly defined, in everyday practice the patients are detected too late. This causes poor local and general prognosis despite intensive treatment. 2. A new concept of "incipient critical ischaemia' is advocated. The leading idea is to detect the risk much earlier and secure prompt adequate therapy. To this goal simple clinical tests of sufficient sensitivity and specificity were elaborated and checked for the use of general practitioners. Any single positivity substantiates the diagnosis of incipient critical ischaemia and transport to a vascular centre for therapy. 3. Evidence is presented showing that with the above approach prognosis is remarkably better and many unnecessary amputations can be prevented. PMID- 8718811 TI - Objectification of compression therapy haemodynamic effect by means of light reflex rheography. AB - Light reflex rheography (LRR) is one of the non-invasive diagnostic methods which makes it possible to examine venous haemodynamics. Through the use of this method we wanted to prove the positive influence of compression therapy on varices of the lower extremities in the 2nd degree chronic insufficiency. In all, 32 patients were examined (39 extremities), and evaluations were done on them of the initial venous refilling time-Ti, i.e. a segment of the LRR curve during the first three seconds after the cessation of muscle work (12 dorsal flexions sitting). Measurements were made without stockings and with the use of the 2nd compression class stockings made by Loana and with the use of Cambren the 1st compression class stockings. With the 1st compression class stockings, measurements were made with one and two stockings simultaneously. The arithmetic mean of the values measured in the individual groups was evaluated by Student's t test. Our results confirmed the anticipated prolongation of the initial time (Ti) segment in the monitored patients, even when the 1st class Cambren C stockings were used. Especially when two of these stockings were used simultaneously, the Ti period was fully comparable to the Ti values when the 2nd class compression stockings were used. It has been demonstrated that when two lower compression class stockings are used simultaneously, there is a significantly increased compression effect and an improvement of the function parameters, even in patients with varices and the 2nd degree chronic venous insufficiency. PMID- 8718812 TI - Homocysteine theory of atherosclerosis. AB - A short survey is presented of the homocysteine theory of atherosclerosis with the key role of endothelial injury. Hyperhomocysteinaemia is a newly emerging risk factor of atherosclerosis. Both hereditary and nutritional influences may contribute to its occurrence. As this risk factor concerns 30-50 percent of atherosclerotics, it may be expected that new methods making possible its detection will be developed and routinely used. Beside the determination of homocysteine blood levels some indicators of endothelial dysfunction such as endothelaemia after methionine load may be used in the future. PMID- 8718813 TI - Toxic and therapeutic properties of oxygen--a review. PMID- 8718814 TI - Changes in the prevalence of cardiovascular diseases risk factors in a group of bank managers (a follow-up study). PMID- 8718816 TI - Body fat assessment by a new bipedal bioimpedance instrument in normal weight and obese women. AB - The aim of the study was to evaluate a new bioimpedance method for assessment of body fat employing bipedal electrodes instead of those attached to both upper and lower extremities. The new analyzer (TBF-105, Tanita Corp., Tokyo, Japan) enables simultaneous measurements of body weight and total body resistance in a subject standing on the stainless steel electrodes. The instrument was tested in both normal weight and obese women. Fat mass estimated by bipedal bioimpedance was highly correlated with that determined by hydrodensitometry (n = 145, r = 0.945, p < 0.001). Fat mass estimated by bipedal bioimpedance significantly correlated not only with subcutaneous fat measured as a sum of 10 skinfolds (r = 0.758, p < 0.001) but also with visceral fat determined as an area on CT scan (r = 0.780, p < 0.001). Anthropometric variables did not substantially influence the differences revealed in fat mass determined by bipedal bioimpedance and by densitometry. An overestimation of total fat mass by bipedal bioimpedance has not been revealed in severely obese individuals, even in those with higher fat accumulation in the limb region. In conclusion, our data have demonstrated that the new bioimpedance instrument employing bipedal electrodes represents a reliable tool for rapid body fat assessment in both normal weight and obese women. PMID- 8718815 TI - The relationship of cardiorespiratory variables, blood lipids, amount and distribution of fat in middle aged men. AB - 68 middle aged men (43.8 +/- 7.4 years) without apparent health problems were examined. Medical check up, individual and family history, physical performance test on a bicycle ergometer, blood lipid level (total, T-, HDL-, LDL-cholesterol, triglycerides) and anthropometric measurements were executed. No serious health problems and complaints were assessed. Average values of body mass index (BMI = 26.7 +/- 3.4 kg/m2 and the percentage of depot fat (20.8 +/- 4.2%) were increased according to standard values. Waist/hip ratio was 0.9 +/- 0.1. Nearly a quarter of men was evaluated as obese. The results of work load test on a bicycle ergometer showed adequate results, however especially the diastolic blood pressure was increased in 11.5% of men at rest, and in an approximately same number of men there appeared the symptoms of cardiac ischemia after the work load. The prevalence of hypertension according to WHO criteria was 9.6%. TCH was 5.35 +/- 1.04, HDL 1.11 +/- 0.19, and triglycerides 1.81 +/- 0.77 mmol/l. Atherogenic index was 4.9 +/- 1.1. Glycemia was 5.04 +/- 0.66 mmol/l. In more than 20% of men there appeared risk values of blood lipids. -Indices evaluated from anthropometric measurements such as waist/hip ratio correlated significantly with blood lipids (positive correlation: total cholesterol, triglycerides, atherogenic index) and/or the level of physical performance (negative correlation), which indicated the possibility to use them as simple markers of cardiovascular risk in larger population samples examined under field conditions. PMID- 8718817 TI - Simple anthropometric measurements--relation to body fat mass, visceral adipose tissue and risk factors of atherogenesis. AB - Abdominal type of obesity is linked to risk factors of atherosclerosis and to metabolic diseases. 216 women, aged 17 to 67 years, mean age 43.1 +/- 0.76, mean weight 108.7 +/- 1.6 kg and mean BMI 39.57 kg/m2, were examined. Body composition was assessed by hydrodensitometry, fat distribution was determined by simple anthropometric measurements (waist circumference, sagittal abdominal diameter at the level L4,5, waist to hip ratio-WHR and waist to thigh ratio-WTR), by computed tomography estimating of visceral fat area and by the ultrasound assessment of visceral fat. Close correlation of visceral fat area determined by CT with the ultrasound assessment of visceral fat was found (r = 0.71). In contrast to WHR, a strong correlation of the waist circumference and sagittal abdominal diameter to total body fat content (r = 0.89, r = 0.84, respectively). Close relation of the waist circumference, sagittal abdominal diameter and WTR to visceral fat examined by ultrasound was discovered (r = 0.61, r = 0.66 and r = 0.58, respectively). The sample was divided into quintiles according to waist circumference, abdominal sagittal diameter and WHR. After adjustment for weight and age, waist circumference and sagittal abdominal diameter tend to be more closely related to the metabolic variables (total cholesterol, HDL-cholesterol, triglycerides, fasting blood glucose, fasting insulin, uric acid) and to the arterial blood pressure than WHR. In the group followed only weak relation was found between anthropometric measures of fat distribution and occurrence of diseases complicating obesity, probably as a result of a high overall fatness connected with a high content of visceral fat. PMID- 8718818 TI - Indices of plasma lipoprotein metabolism in normolipidemic myocardial infarction survivors. AB - BACKGROUND: Many patients suffering from ischaemic heart disease (IHD) were shown to be characterized by different deviations of lipoprotein metabolism without having plasma cholesterol and triglyceride levels increased. The aim of the study was to determine the parameters of lipoprotein metabolism most contributing to finding among normolipidemic men those with an elevated risk of myocardial infarction. METHODS AND RESULTS: We investigated 21 men with normal plasma cholesterol and triglyceride levels, who survived myocardial infarction (MI) in the period 6 months to 3 years before the investigation (group MI). In comparison with the control group of normolipidemic men without IHD, survivors of MI had significantly lower concentrations of HDL-C (-22%, P < 0.01), especially in HDL3 subfraction (-24%, P < 0.001), higher concentrations of apo B in LDL (+36%, P < 0.001) and lower values of cholesterol to apo B ratio in LDL (-25%, P < 0.05). The concentrations of total plasma cholesterol, triglycerides, LDL-cholesterol, HDL2-cholesterol, apo A-I and the ratio HDL-C/apo A-I did not differ significantly. Discriminant analysis revealed that for separating the two groups, determining apo B in LDL and HDL3-C was most advantageous as thus 80% patients in this set could be correctly classified. CONCLUSIONS: These results show the significance of the investigation of apo B and HDL-C including their subfractionism HDL2 and HDL3 for the estimation of MI risk in normolipidemic men. PMID- 8718819 TI - Clinical implications of recognition of the hereditary non-polyposis colon cancer syndrome (HNPCC) for the early detection of colorectal cancer. AB - Hereditary non-polyposis colon cancer (HNPCC), also known as the Lynch syndrome or, previously, as the cancer family syndrome (CFS) is a recently defined autosomal cancer syndrome which is different from familial adenomatous polyposis and accounts for a considerable proportion of colorectal cancer (CRC). The main features of HNPCC include familial aggregation of CRC (either solely or combined with other, particularly gynaecological cancer sites- Lynch I or II, resp.), very frequent localization of the cancer in the right colon, a considerably lower age at the time of occurrence and a high risk of synchronous and metachronous tumours. Recognition of the syndrome through family history offers an important chance to detect CRC at a presymptomatic stage with a positive effect on the prognosis. In 1981/82 we started a prospective follow-up of symptom-free subjects conforming with the criteria of HNPCC. Up to 1994 we found 364 subjects of this type coming from 38 families. After initial colonoscopy these subjects entered a system of colonoscopic surveillance built up according to the individual degree of risk. Initial colonoscopy revealed cancer in 61 and adenomas in 122 subjects, both in more than 70% in the right colon. The mean age at this time was 47.5 in cancer and 45.3 in adenomas. More cancers and adenomas were found on repeated colonoscopies during surveillance. All the cancers were well resectable, a 5 year survival in all. During this course in a group of 239 non-screened symptomatic cases of the HNPCC syndrome, detected only at the time of colonoscopy, cancer was found in 146 and adenomas in 71 patients. Five year survival was 65% in this group. The cancers in the screened group were Dukes A and B (mostly A) in 82% and C in 18%, in the symptomatic group Dukes A or B (mostly B) in 67%, C in 18 and D in 15%. - CONCLUSIONS: The results show more favourable colonoscopic findings and better prognosis in symptom-free, colonoscopally screened cases of HNPCC than in symptomatic ones and fully justify an active approach based on colonoscopic surveillance of HNPCC subjects prior to diagnosis. PMID- 8718820 TI - The development of the digestive endoscopy in Czech Republic. PMID- 8718822 TI - Temporal control mechanism in equaled interval tapping. AB - Subjects who were at intermediate levels of musical performance made equaled interval tapping in several tempos. The temporal fluctuation for the tapping was observed and analysed. The power spectrum of the fluctuation showed a critical phenomenon at around a frequency which corresponds to the period of 20 taps, for all tempos and all subjects, i.e., the slope of the spectrum was flat or had a positive value in the high frequency region above the critical frequency but it increased as the frequency decreased in the low frequency region below the critical frequency. Moreover, auto-regressive models and Akaike's information criterion were introduced to determine the critical tap number. The order of the best auto-regressive model for the temporal fluctuation data was distributed around 20 taps. These results show that the memory capacity of 20 taps governs the control of equaled interval tapping. To interpret the critical phenomenon of 20 taps with the memory capacity of the short term memory, the so called magic number seven, a simple chunking assumption was introduced; subjects might have unconsciously chunked every three taps during the tapping. If the chunking assumption is true, when subjects consciously chunk every seven taps, the memory capacity of taps should shift to about 50 taps. To test if the assumption is true or not, subjects made a three-beat rhythm tapping and a seven-beat rhythm tapping with equaled intervals. As a result, the memory capacity for these accented tappings were also estimated as 20 taps. This suggests that the critical phenomenon cannot be explained by the chunking assumption and the magic number seven, rather this finding suggests that there exists a memory capacity of 20 taps and this is used for equaled interval tapping. PMID- 8718821 TI - The relative utility of health-related fitness tests and skilled motor performance tests as measures of biological age in Japanese men. AB - In the present paper we report the results of a study in which we compared 2 different approaches to the computation of biological age (BA) in a sample of 322 Japanese men (age range 20 to 79 years). In the first approach, 4 commonly used measures of health-related fitness (VO2peak, trunk flexion from a standing position, body fat, and grip strength) were reduced to a single BA score (HRF Age) using principal component analysis. In contrast, in the second approach, 3 commonly used measures of skilled motor performance and agility (vertical jump, stepping side-to-side, and balancing on one leg with eyes closed) were reduced to a single BA score (SMP Age) using similar multivariate procedures. The criterion related validity of both of the BA measures was examined by assessing each measure's ability to discriminate between healthy and active groups of subjects. This was achieved by classifying the original subject pool into regularly active (ACT; n = 108) and healthy (HLTH; n = 169) subgroups on the basis of self reported activity levels. Analyses revealed that HRF Age was a more powerful discriminator between the two activity groups than SMP Age. While HRF Age of HLTH subjects was very close to their chronological age (CA), in the ACT group, HRF Age was on average 15 years less than their CA (P < 0.05). In a separate analysis, we assessed the HRF Age of patients with ischemic heart disease, hypertension, obesity, or diabetes (PAT; n = 45). The HRF Age of these subjects averaged 10 years above their CA. Our data suggest that commonly used measures of health-related fitness can be usefully employed as indices of BA which differentiate between individuals of similar ages but differing health and physical activity status. In contrast, measures of skilled motor performance were found to be less valuable measures of BA. The implication of our findings for future experimental design in exercise and aging research is discussed. PMID- 8718823 TI - Effect of endurance training under hypoxic condition on oxidative enzyme activity in rat skeletal muscle. AB - The adaptive response of oxidative enzyme activity in the skeletal muscle to training in normoxic and in normobaric hypoxic training was studied. Forty male Wistar rats were divided into 4 groups: normoxia + sedentary (NS, n = 10); hypoxia + sedentary (HS, n = 10); normoxia + training (NT, n = 10); and hypoxia + training (HT, n = 10). Rats in the NT group ran on a treadmill for 30 min a day at 20-30 m.min-1, 4 days a week for 10 weeks in normoxia. Rats in the HT group performed the same training protocol as NT in an ambient FIO2 decreased to 12%. HS rats were exposed to hypoxia in the same degree, duration and frequency as HT without exercise. After the training period, the soleus and the plantaris muscles were removed, and the activities of mitochondrial enzymes, malate dehydrogenase (MDH) and 3-hydroxyacyl-CoA dehydrogenase (HAD) were measured by a spectrophotometer. The normoxic training did not increase MDH or HAD activities, in either the soleus or the plantaris. This absence of change in mitochondrial enzyme activities is considered to be the results of inadequate stimulus of training, including a relatively low amount of exercise. On the other hand, the hypoxic training enhanced the MDH activity in the soleus by 17.5% compared with NS (P < 0.01) and by 20.5% compared with HS (P < 0.01). Also in the plantaris, the MDH activity in HT was higher than that in HS (15.7%, P < 0.05). These findings suggest that even moderate training by which enzyme activity is not increased under normoxic conditions can enhance the oxidative capacity in the skeletal muscle when the training is performed in a hypoxic environment. PMID- 8718825 TI - Seasonal changes of salivary immunoglobulin A under the influences of two types of clothing. PMID- 8718824 TI - Changes in aerobic capacity and coronary risk factors during long-term exercise training in women with ischemic heart disease: a 36-month follow-up. AB - We evaluated the time course of alteration in aerobic capacity and coronary risk factors associated with a 36-month exercise program in women with ischemic heart disease (IHD). Twenty-one patients participated in supervised exercise and home based exercise programs for 36 months. However, of all patients, 11 patients completed the entire program. Oxygen uptake corresponding to lactate threshold (VO2LT), peak oxygen uptake (VO2peak), percent of body fat (%BF), systolic (BPs) and diastolic (BPd) blood pressure, total cholesterol (TC), low-density lipoprotein cholesterol (LDLC), high-density lipoprotein cholesterol (HDLC) and triglycerides (TG) were assessed before and 4, 8, 12, 24, and 36 months after exercise. The intensity of exercise was set at individually determined LT, i.e., 60 to 70% VO2peak. The daily amount of exercise during 36 months averaged 24.0 +/ 11.4 minutes per day. During the course of exercise program, VO2LT, VO2peak, BPs, BPd, and TG improved significantly at month 4. Although %BF decreased significantly at month 8 to 12, it tended to return to the initial level at month 36. On the other hand, a significant increase in HDLC was found at month 24 and improved state of HDLC remained unchanged thereafter. No changes were found in TC and LDLC. These results suggest that most of the beneficial effects of exercise on aerobic capacity and coronary risk factors in women with IHD are obtained within 4-8 months, with some further improvement seen with continued exercise up to 24 months. PMID- 8718826 TI - Food safety. PMID- 8718827 TI - Solving educational gaps. PMID- 8718828 TI - Ethical checklist for dental practice. PMID- 8718829 TI - The changing marketplace: informed choices; safe action. PMID- 8718830 TI - The changing marketplace: informed choices; safe action. PMID- 8718831 TI - Changing the law: how boards of dental examiners influence the legislative process. PMID- 8718832 TI - Initial licensure in dentistry: how boards of dental examiners test candidates for licensure. PMID- 8718833 TI - A voice in national policy: how boards of dental examiners affect education and practice. PMID- 8718834 TI - License without examination, the Washington state experience: how boards of dental examiners might alternatively license dentists. PMID- 8718835 TI - Two sides to every story: how state boards of examiners enforce disciplinary actions. PMID- 8718836 TI - The form and function of a practice: how state boards of examiners affect dental practice. PMID- 8718837 TI - Dentists at war. PMID- 8718838 TI - Strategic planning. PMID- 8718839 TI - Distance education in nursing. AB - Although not for everyone, distance education is a "connecting point" for faculty and students who are separated by time and space. As technology becomes increasingly available to nurse educators, the instructional and public relations advantages become significant benefits to nurse educators. PMID- 8718840 TI - Telenursing, virtual nursing, and beyond. Definition. PMID- 8718841 TI - Modems: the gateway to cyberspace. PMID- 8718842 TI - Taking a bite out of CRISP. Strategies on using and conducting searches in the Computer Retrieval of Information on Scientific Projects database. AB - A vast array of information useful to the scientific community is readily available on the internet. Currently, the National Institutes of Health (NIH) offers user access to several large databases, some of which are pertinent to nurses interested in research, health policy formation, and identifying funded research projects. One such database, the Computer Retrieval of Information on Scientific Projects (CRISP), provides information on research grants funded by the NIH from 1972 to the present. Retrieval of scientific information for each project in CRISP is made available by project title, grant number, abstract, principal investigator, eight-digit CRISP Thesaurus terms, and key words. This article introduces the reader to CRISP and provides a set of succinct strategies for conducting comprehensive searches in the database. PMID- 8718843 TI - Nurses' computer-mediated communications on NURSENET. A case study. AB - Computer-mediated communication (CMC) provides a new medium through which large numbers of nurses who might not have other opportunity can "meet" and exchange information and ideas. This article describes methodologic issues and presents findings arising from a case study, including a qualitative discourse analysis, of information exchange and discussion on the NURSENET list server. A 2-day sample of list discussions was analyzed using Fairclough's discourse analysis method, and a small number of subscribers were interviewed via electronic-mail. These methods, as opposed to purely quantitative ones, seem increasingly appropriate to the analysis of CMC. Findings from the study included an over representation of males in discussions relative to their proportion of subscribers; an absence of the more antisocial on-line behavior (flaming) seen in many discussion lists; and some suggestion that CMC users challenge traditional nursing discourse. This article presents research findings previously available only on-line. PMID- 8718844 TI - End-user satisfaction of a patient education tool manual versus computer generated tool. AB - This article reports a nonexperimental comparative study of end-user satisfaction before and after implementation of a vendor supplied computerized system (Micromedex, Inc) for providing up-to-date patient instructions regarding diseases, injuries, procedures, and medications. The purpose of this research was to measure the satisfaction of nurses who directly interact with a specific patient educational software application and to compare user satisfaction with manual versus computer generated materials. A computing satisfaction questionnaire that uses a scale of 1 to 5 (1 being the lowest) was used to measure end-user computing satisfaction in five constructs: content, accuracy, format, ease of use, and timeliness. Summary statistics were used to calculate mean ratings for each of the questionnaire's 12 items and for each of the five constructs. Mean differences between the ratings before and after implementation of the five constructs were significant by paired t test. Total user satisfaction improved with the computerized system, and the computer generated materials were given a higher rating than were the manual materials. Implications of these findings are discussed. PMID- 8718845 TI - Creating a nursing home page on the World Wide Web. AB - The authors provide a brief overview of the internet and home pages on the World Wide Web. Definitions of Web terminology are provided to help the reader understand home page creation. The authors also describe the steps in electronic publishing and how to create a home page. Supplemental tables provide internet addresses for nursing and non-nursing sites for reviewal of other home pages. The article continues with information about design, formatting, adding text and images, and dissemination suggestions for home pages. Examples of home pages and instruction commands (tags) are provided. The future of Web publishing is discussed, and issues and concerns are raised regarding electronic publishing. PMID- 8718846 TI - Biogenesis of polytopic membrane proteins: membrane segments of P-glycoprotein sequentially translocate to span the ER membrane. AB - The initial steps in the biogenesis of membrane proteins parallel those of secretory proteins. However, membrane proteins contain a signal to stop translocation across the membrane. For polytopic membrane proteins, those with multiple transmembrane segments, little is known of the temporal sequence or relationship between synthesis of the nascent proteins, translocation, folding, and integration of the membrane segments into the bilayer. Here we demonstrate that latent membrane segments translocate sequentially as they emerge from the ribosome and do not accumulate on the cytosolic side to form loops, or larger structures, prior to translocation across the membrane. PMID- 8718848 TI - Autocatalyzed protein folding. AB - Proline isomerization, an intrinsically slow process, kinetically traps intermediates in slow protein folding reactions. Thus, enzymes that catalyze proline isomerization (prolyl isomerases) often catalyze protein folding. We have investigated the folding kinetics of FKBP, a prolyl isomerase. The main conclusion is that FKBP catalyzes its own folding. Altogether, the FKBP refolding kinetics are resolved into three exponential phases: a fast phase, tau 3; an intermediate phase, tau 2; and a slow phase, tau 1. Unfolding occurs in a single phase, the unfolding branch of phase tau 2. In the presence of native FKBP, both the intermediate (tau 2) and slow (tau 1) phases are faster, suggesting that folding phases tau 1 and tau 2 involve proline cis-trans isomerization. In the absence of added native FKBP, autocatalytic folding of FKBP is detected. For refolding starting with all the FKBP unfolded initially, the slowest folding phase (tau 1) is almost 2-fold faster at a final concentration of 14 microM FKBP than at 2 microM FKBP, suggesting that catalytically active FKBP formed in the fast (tau 3) or intermediate (tau 2) folding phases catalyzes the slow folding phase (tau 1). Moreover, autocatalysis of folding is inhibited by FK506, an inhibitor of the FKBP prolyl isomerase activity. The results show that the slow phase in FKBP folding is an autocatalyzed formation of native FKBP from kinetically trapped species with non-native proline isomers. While the magnitude of the catalytic effects reported here are modest, FKBP folding may provide a prototype for autocatalysis of kinetically trapped macromolecular conformational changes in other systems. PMID- 8718847 TI - Effects of protein stabilizing agents on thermal backbone motions: a disulfide trapping study. AB - Chemical stabilizers are widely used to enhance protein stability, both in nature and in the laboratory. Here, the molecular mechanism of chemical stabilizers is studied using a disulfide trapping assay to measure the effects of stabilizers on thermal backbone dynamics in the Escherichia coli galactose/ glucose binding protein. Two types of backbone fluctuations are examined: (a) relative movements of adjacent surface alpha-helices within the same domain and (b) interdomain twisting motions. Both types of fluctuations are significantly reduced by all six stabilizers tested (glycerol, sucrose, trehalose, L-glucose, D-glucose, and D galactose), and in each case larger amplitude motions are inhibited more than smaller ones. Motional inhibition does not require a high-affinity stabilizer binding site, indicating that the effects of stabilizers are nonspecific. Overall, the results support the theory that effective stabilizing agents act by favoring the most compact structure of a protein, thereby reducing local backbone fluctuations away from the fully folded state. Such inhibition of protein backbone dynamics may be a general mechanism of protein stabilization in extreme thermal or chemical environments. PMID- 8718849 TI - Structural change in alpha-chymotrypsin induced by complexation with alpha 1 antichymotrypsin as seen by enhanced sensitivity to proteolysis. AB - Both human neutrophil elastase (HNE) and free chymotrypsin (Chtr) proteolyze Chtr within the complex that Chtr forms with antichymotrypsin (ACT). As free Chtr is stable both to self-digestion and to digestion by HNE, these results are indicative of a stability and/or conformational change in Chtr that accompanies complex formation. As determined by both N-terminal sequence analysis and matrix assisted laser desorption ionization mass spectroscopy (MALDI-MS), the major initial sites of HNE cleavage of complexed Chtr are between gamma-chain residues A158/S159 and V188/S189. Significantly, this latter site is at the base of the S1 site that recognizes the P1 position of the serpin. A slower cleavage in the beta chain between T139/G140 is also found. In addition, rACT is cleaved between residues V22/D23. The gamma-chain of complexed Chtr is also cleaved by free Chtr, but at different sites: L162/L163 and W172/G173. beta-Chain cleavages were also found between residues Q81/K82 and F114/S115. Cleavages similar to those described above were also found when Chtr was complexed with the L358F-rACT variant, but not for Chtr complexed with either of the smaller inhibitors bovine pancreatic trypsin inhibitor or turkey ovomucoid third domain, nor for the covalent adduct of Chtr with N-p-tosylphenylalanyl chloromethyl ketone. We conclude that the structural change in Chtr making it a proteinase substrate is coupled with the large conformational change in ACT following complex formation. Complexed Chtr is much less reactive toward proteolytic digestion in the presence of high salt than in its absence, in accord with the high-salt induced release of active enzyme from the Chtr.rACT complex and the suggestion that electrostatic interactions mediate the coupling of structural change between rACT and Chtr within the Chtr.rACT complex. Potential physiological consequences of this work are explored. PMID- 8718850 TI - Structures of the Klebsiella aerogenes urease apoenzyme and two active-site mutants. AB - Urease from Klebsiella aerogenes [Jabri et al. (1995) Science 268, 998-1004] is an (alpha beta gamma)3 trimer with each alpha-subunit having an (alpha beta)8 barrel domain containing a binickel active center. Here we examine structure function relations for urease in more detail through structural analysis of the urease apoenzyme at 2.3 A resolution and mutants of two key catalytic residues (H219A and H320A) at 2.5 A resolution. With the exception of the active site, in which a water molecule takes the place of the missing carbamate and nickel atoms, the structure of the apoenzyme is nearly identical to that of the holoenzyme, suggesting a high degree of preorganization which helps explain the tight binding of nickel. In the structure of H219A, the major change involves a conformational shift and ordering of the active site flap, but a small shift in the side chain of Asp alpha 221 could contribute to the lower activity of H219A. In the H320A structure, the catalytic water, primarily a Ni-2 ligand in the holoenzyme, shifts into a bridging position. This shift shows that the nickel ligation is rather sensitive to the environment and the change in ligation may contribute to the 10(5)-fold lower activity of H320A. In addition, these results show that urease is resilient to the loss of nickel ions and mutations. Analysis of the urease tertiary/quaternary structure suggests that the stability of this enzyme may be largely due to its burial of an unusually large fraction of its residues: 50% in the gamma-subunit, 30% in the beta-subunit, and 60% in the alpha-subunit. PMID- 8718851 TI - Crystal structures of complexes of a peptidic inhibitor with wild-type and two mutant HIV-1 proteases. AB - Crystal structures of the protease of human immunodeficiency virus type 1 (HIV-1) and two mutant proteases, V82D and V82N, have been determined. In all three cases the enzyme forms a complex with the peptidic inhibitor U-89360E. All structures have been determined to 2.3 A resolution and have satisfactory agreement factors: 0.173 for wild type, 0.175 for V82D, and 0.182 for V82N. Comparison of the three crystal structures provides explanations which are consistent with the known kinetic properties of these mutant enzymes with the U-89360E inhibitor [Lin, Y., Lin, X., Hong, L., Foundling, S., Heinrikson, R. L., Thaisrivongs, S., Leelamanit, W., Raterman, D., Shah, M., Dunn, B.M., & Tang, J. (1995) Biochemistry 34, 1143-1152]. Unfavorable van der Waals interactions between the inhibitor and the mutated side chains at position 82 are consistent with diminished affinity for the inhibitor by the mutant enzymes. If a mutation is potentially resistant to an inhibitor, the mutant enzyme should not only have an increased Ki for the inhibitor but should also preserve considerable catalytic capability. The V82D mutant possesses these qualities. In the V82D crystal structure, a water molecule, which connects the protease flap to the inhibitor, is missing or of low occupancy. Absence of this bridge may be important in determining catalytic capability. Moreover, mutation at position 82 induces change in two polypeptide backbone regions, 35-41 and 67-68, which may be related to protease flap mobility. PMID- 8718852 TI - Rational design of specific high-affinity peptide ligands for the Abl-SH3 domain. AB - SH3 domains bind proline-rich peptides with affinities in the order of 0.2-50 microM. In general, these domains are quite promiscuous, and the same peptide can bind to several different SH3 domains with similar affinities (i.e., 3BP1 peptide to Abl- and Fyn-SH3). This poor affinity and specificity make it difficult to elucidate their role in vivo as well as the use of peptides to specifically bind to a single domain. Here, we report that by using existing biocomputing tools, as well as simple physicochemical reasoning, it is possible to design mutations in the 3BP1 peptide (Met4-Tyr, Pro5-Ser, and Leu8-Pro), so that the affinity for Abl SH3 increases 20-fold (p40 peptide: APTYSPPPPP; Kd = 0.4 microM), while that for the closely related domain, Fyn-SH3, decreases 10-fold. Both the RT and n-Src loops are responsible for regulating the specificity for Pro-rich ligands and more specifically residues Ser15, Thr19, and Glu38 in Abl-SH3. The first six positions in the 3BP1 peptide are important for determining the specificity for SH3 domains, while the remaining four seem to be more important for the affinity. Moreover, by choosing rationally the substituents, it is possible to replace some of the Pro residues postulated to be essential for the interaction with SH3 domains and still have a significant affinity. This indicates that the sequence repertoire that could interact with a specific SH3 domain could be larger than previously thought. PMID- 8718853 TI - Synthesis and characterization of leiurotoxin I analogs lacking one disulfide bridge: evidence that disulfide pairing 3-21 is not required for full toxin activity. AB - Leiurotoxin I (Lei-NH2), a toxin isolated from the venom of the scorpion Leiurus quinquestriatus hebraeus, is a blocker of the apamin-sensitive Ca(2+)-activated K+ channels. It is a 31-residue polypeptide cross-linked by three disulfide bridges which are presumably between Cys3-Cys21, Cys8-Cys26, and Cys12-Cys28. To investigate the role of these disulfides, analogs of Lei-NH2 lacking one disulfide bridge (i.e., [Abu3,21]Lei-NH2, [Abu8,26]Lei-NH2, and [Abu12,28]Lei NH2) were chemically synthesized by selective replacement of each pair of half cystines forming a bridge by two alpha-aminobutyrate (Abu) residues. The two disulfide pairings of the main folded form of the synthetic analogs were established by enzymatic proteolysis. They were as expected between Cys8-Cys26 and Cys12-Cys28 for [Abu3,21]Lei-NH2 but were unexpectedly between Cys3-Cys12 and Cys21-Cys28 for [Abu8,26]Lei-NH2 and between Cys3-Cys8 and Cys21-Cys26 for [Abu12,28]Lei-NH2. The synthetic peptides were tested in vitro for their capacity to compete with the binding of [125I]apamin to rat brain synaptosomes and in vivo for their neurotoxicity in mice. In both assays, [Abu3,21]Lei-NH2 exhibited full Lei-NH2-like activity whereas [Abu8,26]Lei-NH2 and [Abu12,28]-Lei-NH2 possessed only residual activities (< 2% native toxin activity). This suggests that disulfide bridge Cys3-Cys21 is not essential per se for high toxin activity. Circular dichroism (CD) spectroscopy of the three analogs showed that only [Abu3,21]Lei-NH2 exhibited a CD spectrum similar to that of Lei-NH2, suggesting they both adopt closely related conformations, in agreement with the pharmacological data. Structural models of the analogs were constructed on the basis of the disulfide pairing assignment and compared with that of Lei-NH2. PMID- 8718854 TI - Crystal structure of thermostable family 5 endocellulase E1 from Acidothermus cellulolyticus in complex with cellotetraose. AB - The crystal structure of the catalytic domain of the thermostable endocellulase E1 from Acidothermus cellulolyticus in complex with cellotetraose has been solved by multiple isomorphous replacement and refined at 2.4 A resolution to an R factor of 0.18 (Rfree = 0.24). E1cd is a member of the 4/7 superfamily of hydrolases, and as expected, its structure is an (alpha/beta)8 barrel, which constitutes a prototype for family 5-subfamily 1 cellulases. The cellotetraose molecule binds in a manner consistent with the expected Michaelis complex for the glycosylation half-reaction and reveals that all eight residues conserved in family 5 enzymes are involved in recognition of the glycosyl group attacked during cleavage. Whereas only three residues are conserved in the whole 4/7 superfamily (the Asn/Glu duo and the Glu from which the name is derived), structural comparisons show that all eight residues conserved in family 5 have functional equivalents in the other 4/7 superfamily members, strengthening the case that mechanistic details are conserved throughout the superfamily. On the basis of the structure, a detailed sequence of physical steps of the cleavage mechanism is proposed. A close approach of two key glutamate residues provides an elegant mechanism for the shift in the pKa of the acid/base for the glycosylation and deglycosylation half-reactions. Finally, purely structural based comparisons are used to show that significant differences exist in structural similarity scores resulting from different methods and suggest that caution should be exercised in interpreting such results in terms of implied evolutional relationships. PMID- 8718855 TI - Active site mutants of pig citrate synthase: effects of mutations on the enzyme catalytic and structural properties. AB - We examined the catalytic efficiency of 18 pig citrate synthase mutants. The residues mutated were selected according to two criteria: the conservation of that residue in all known citrate synthase sequences, and the importance of that residue in substrate-amino acid interactions suggested by the extensive crystal structure information on the enzyme and its complexes. Several changes were made at certain residues to probe the effects of size, hydrogen bonding, and charge on the kinetics of the enzyme. The mutations, as expected, affected the kcats and Kms for OAA and acetyl-CoA to varying degrees. The catalytic efficiency of each of the mutants was determined by the kcat/Km for the individual substrates, OAA and acetyl-CoA. All mutations affected kcat. There was only one mutant, Asp327 Asn, in which the Kms primarily were affected. Most mutations affected both kcat and Km and included the following: His274Gly, His274Arg, Asp375Gly, Asp375Asn, Asp375Glu, Asp375Gln, His320Gly, His320Gln, His320Asn, His320Arg, Arg401His, Gly275Val, and Gly275Ala. The mutations, Arg401Gly, Arg401Lys, His235Gln, and Asn242Glu, had smaller effects on kcat and Km. The CS mutant Arg401Lys exhibited a modestly improved kcat/Km for both substrates compared to the nonmutant enzyme. X-ray crystallographic studies at 2.7 A resolution of one of the mutants, His274Gly, have been undertaken. The mutant enzyme crystallizes in an "open" conformation essentially isomorphous to wild type. The refined model has good geometry and a crystallographic R factor of 0.187 for 11 441 reflections observed between 6.0 and 2.7 A resolution. The refined model revealed a localized relaxation of the structure to relieve strain imposed by a high-energy main and side chain conformation of His274 in the nonmutant, but otherwise the mutation does not result in major structural alterations. Preliminary electrostatic calculations provide support for the concept that the transition state in the rate-limiting step of the citrate synthase catalyzed reaction may be an "enolized" version of acetyl-CoA that is neither neutral nor fully negatively charged and that a possible role for the catalytically essential His274 is to stabilize this by charge delocalization mediated by a hydrogen bond. These results provide the basis for further studies of the effects of these changes on the several reactive intermediates, activated substrates, and transition states which may occur along the reaction coordinate for this type of Claisen enzyme. PMID- 8718856 TI - Biochemical evidence that a D-loop is part of a four-stranded PNA-DNA bundle. Nickel-mediated cleavage of duplex DNA by a Gly-Gly-His bis-PNA. AB - A peptide nucleic acid (PNA) with improved strand-displacement capability and a site-specific DNA cleavage function is a novel reagent for probing the structure of PNA-DNA complexes in solution. By linking two PNAs in tandem with an aliphatic linker, the bis-PNA forms a bis-PNA-DNA triple-stranded complex having a higher stability to thermal denaturation than conventional monomeric PNAs. When a Gly Gly-His tripeptide is placed on either the Watson-Crick or Hoogsteen bis-PNA strand, nickel-mediated cleavage is detected at specific sites on the displaced and hybridized DNA strands. Because the displaced strand is cleaved when GGH is placed on either PNA strand, the D-loop must be close to the backbone of the bis PNA-DNA triplex. Furthermore, the pattern of cleavage on the displaced strand suggests the nickel-tripeptide complex lies in a groove formed by the displaced DNA strand and both PNA strands. These observations suggest that the D-loop is a part of a four-stranded bis-PNA-DNA2 bundle. PMID- 8718857 TI - Biogenesis, cellular localization, and functional activation of the heat-stable enterotoxin receptor (guanylyl cyclase C). AB - Enterotoxigenic Escherichia coli elaborate a peptide called heat-stable enterotoxin (ST), which binds to and activates the intestinal ST receptor (STaR). STaR, also known as guanylyl cyclase C (GC-C), is a member of the transmembrane guanylyl cyclase receptor family. The mRNA for STaR encodes an approximately 120 kDa protein with the N-terminal ligand binding domain on the cell surface. Ligand affinity cross-linking studies have previously demonstrated several species of potential ST binding proteins, ranging in size from approximately 50 to 160 kDa. Although these smaller forms of STaR (50-80 kDa) have been proposed to act in vivo as toxin binding proteins, their biogenesis and localization have not previously been examined. Using pulse labeling in vivo and synchronized translation in vitro, we demonstrate that these smaller forms represent incomplete translational products and are not formed through limited proteolysis of the full-length receptor, as had previously been believed. We determined, using fluorescence confocal microscopy and surface labeling, that only approximately 25% of cellular receptors are expressed at the surface, while the remaining population is retained within the endoplasmic reticulum. Only full length receptor is found at the surface of the cell, indicating this to be the biologically active form of STaR responsible for interacting with the heat-stable enterotoxin and other luminal intestinal peptides. The large intracellular receptor population, and potential for function before translocation to the cell surface, may impact on how pharmacologic modulators of this clinically important receptor are designed. PMID- 8718858 TI - Crystal structure of soybean lipoxygenase L-1 at 1.4 A resolution. AB - Lipoxygenases, which are widely distributed among plant and animal species, are Fe-containing dioxygenases that act on lipids containing (Z,Z)-pentadiene moieties in the synthesis of compounds with a variety of functions. Utilizing an improved strategy of data collection, low temperature, and synchrotron radiation of short wavelength, the structure of ferrous soybean lipoxygenase L-1, a single chain protein of 839 amino acid residues, has been determined by X-ray crystallography to a resolution of 1.4 A. The R-factor for the refined model is 19.7%. General features of the protein structure were found to be consistent with the results of prior crystallographic studies at lower (2.6 A) resolution. In contrast to the prior studies, the binding of a water molecule to the active site Fe was established. The octahedral coordination sphere of the Fe also includes the side chains of His499, His504, His690, and Asn694 as well as the terminal carboxylate of Ile839, which binds as a monodentate ligand. Asn694 is involved in a number of labile polar interactions with other protein groups, including an amide-aromatic hydrogen bond, and appears to be a weak ligand. Several possible access routes for dioxygen and fatty acids to the internal active site and substrate binding cavity are described. The protein structure restricts access to the Fe site such that the formation of an organo-Fe intermediate seems improbable. Structural restrictions pertinent to other proposed reaction intermediates, such as planar pentadienyl and nonplanar allyl radicals, are also discussed. PMID- 8718859 TI - Structure of 3 alpha-hydroxysteroid/dihydrodiol dehydrogenase complexed with NADP+. AB - Rat liver 3 alpha-hydroxysteroid/dihydrodiol dehydrogenase (3 alpha-HSD) inactivates circulating steroid hormones and is involved in polycyclic aromatic hydrocarbon (PAH) carcinogenesis. It is the only HSD of known structure in the aldo-keto reductase (AKR) superfamily and may provide a paradigm for other mammalian HSDs in this family. The structure of the 3 alpha-HSD.NADP+ binary complex has been determined at 2.7 A resolution and refined to a crystallographic R-factor of 23.4% with good geometry. The model is similar to other binary complexes in the AKR superfamily in that NADP+ binds at the C-terminal end of an alpha/beta barrel. However, it is unique in that NADP+ is bound in two alternate conformations, probably because of the lack of a salt-linked "safety belt" over the pyrophosphate bridge. The structure supports a previously proposed catalytic mechanism for carbonyl reduction in which Tyr 55 is the general acid, and its effective pKa is lowered by the adjacent Lys 84. We present evidence that the structurally distinct short-chain dehydrogenase/reductase (SDR) superfamily may have convergently evolved a similar catalytic mechanism. Insight into substrate binding is offered by a crystal packing contact in which a neighboring molecule inserts a tryptophan residue (Trp 227) into an apolar cleft in 3 alpha-HSD. This cleft is proximal to the bound NADP+ cofactor and contains a surface of apolar residues (Leu 54, Trp 86, Leu 122, Phe 128, Phe 129, Leu 137, Phe 139), making it a likely candidate for the substrate-binding site. Thus, in forming this crystal contact, Trp 227 may mimic a portion of a bound steroid. In addition, we propose that a water molecule in the active site indicates the position of the hydroxyl oxygen in a 3 alpha-hydroxysteroid substrate. Knowledge of the position of this water molecule, combined with the stereochemistry of hydride transfer, suggests that the alpha face of a bound steroid will be oriented toward the side of the apolar cleft containing Trp 86. PMID- 8718860 TI - High-efficiency triple-helix-mediated photo-cross-linking at a targeted site within a selectable mammalian gene. AB - Targeting damage to specific sites in the genome represents an attractive approach to manipulating gene function in mammalian cells. To test the applicability of triple-helix formation as a means for achieving precisely timed site-specific damage within a mammalian gene, a triplex-forming oligodeoxyribonucleotide (TFO) that binds with high affinity to a specific site within the hamster adenine phosphoribosyltransferase (APRT) gene was modified with the photochemically reactive psoralen derivative 4'-(hydroxymethyl)-4,5',8 trimethylpsoralen (HMT). The modified TFO, psorTFO1, bound with high affinity to a target site within intron 1 of the APRT gene. Upon irradiation, photomonoadducts (i.e., covalent adducts of psorTFO1 to one strand of the target duplex) were formed with high efficiency (approximately 50%). Introduction of 5' TpA sequences (the preferred site for psoralen-induced photo-cross-links) at or near the triplex junction leads to increased efficiency of total photoadduct formation and to efficient formation of products that had the electrophoretic mobility on denaturing PAGE expected for three-stranded photo-cross-links (i.e., products containing psorTFO1 covalently linked to both strands of the duplex). Their identities as cross-links were verified by (1) identical electrophoretic mobility of products formed with either duplex strand radiolabeled and (2) coprecipitation of the radiolabeled duplex strand with its complementary biotinylated strand following denaturation. In addition, the cross-links were completely reversible upon irradiation at 254 nm, as expected for psoralen mediated cross-links. The yield and distribution of photoadducts depended on 5' TpA position. The most efficient photoadduct formation (approximately 90%) and photo-cross-link formation (approximately 90% of total photoadducts) were observed for a 5'-TpA adjacent to the triplex junction, with significant, but lower, cross-linking efficiency within three base pairs of the junction. Molecular models of the psoralen-conjugated triplex with its six-carbon linker suggested a simple explanation for this distance dependence: facile intercalation near the triplex/duplex junction, with increasing strain required for intercalation at more distant sites. These results indicate that psorTFO1 allows for DNA damage with high precision and high efficiency, and the likely proportion of monoadducts and cross-links can be estimated from the target sequence. PMID- 8718861 TI - The importance of the second hairpin loop of cystatin C for proteinase binding. Characterization of the interaction of Trp-106 variants of the inhibitor with cysteine proteinases. AB - The single Trp of human cystatin C, Trp-106, is located in the second hairpin loop of the proteinase binding surface. Substitution of this residue by Gly markedly altered the spectroscopic changes accompanying papain binding and reduced the affinity for papain, actinidin, and cathepsins B and H by 300-900 fold. The decrease in affinity indicated that the side chain of Trp-106 contributes a similar free energy, -14 to -17 kJ.mol-1, to the binding to all four cysteine proteinases, corresponding to about 20-30% of the total binding energy. Replacement of Trp-106 by Phe led to a smaller (30-120-fold) decrease in affinity for the four enzymes than Gly substitution. The binding energy of the Phe residue corresponded to 20-45% of that of Trp, showing that a phenyl group can only partly substitute for the indole ring. The reduced affinities of the cystatin C Trp-106 variants for all proteinases studied were due almost exclusively to increased dissociation rate constants. The second hairpin loop thus contributes to the binding primarily by keeping cystatin C anchored to the proteinase once the complex has been formed. This role is partly in contrast to that of the N-terminal region, which increases the affinity of cystatin C for cathepsin B by increasing the association rate constant. Removal of the N terminal region of the Trp-106-->Gly variant by proteolytic cleavage substantially weakened the binding to papain and cathepsin B. The resulting affinity indicated that the first hairpin loop (the "QVVAG-region"), which is the only region of the proteinase binding surface remaining intact in the truncated variant, contributes 40-60% of the total free energy of binding of cystatin C to both proteinases. PMID- 8718862 TI - Cleavages induced by different metal ions in yeast tRNA(Phe) U59C60 mutants. AB - The U59 and C60 residues, which form the strong Pb(II) ion binding site in yeast tRNA(Phe), were systematically mutated, and the effects of individual mutations on cleavages induced by various metal ions were analyzed. It turned out that the presence of C60 is essential for efficient cleavage of the D-loop by Pb(II), Eu(III), and Mg(II)ions. On the other hand, manganese ions were capable of cleaving these mutants with an efficiency similar to that observed for the wild type transcript. Moreover, in all C60 mutants, the main Mn(II) cleavage site was shifted by one phosphate from P16 to P17. Mutations of U59 did not affect so dramatically the efficiency and specificity of the D-loop hydrolysis induced by all studied metal ions. In the G59C60 mutant cleaved by Pb(II) ions, new cuts took place in the T-stem at P63-P65. Also, the C60 mutants were cleaved more strongly at P49 by Pb(II) ions. In G59C60 and A59C60 as well as in all C60 mutants, the Mg(II) and Mn(II) cleavage at P61 was suppressed. Nevertheless, the changes in overall tRNA structure resulting from U59 and C60 mutations were rather subtle. The studied mutants showed S1 and V1 nuclease digestion patterns practically indistinguishable from those observed in the wild type transcript. The metal ions are shown to be well-suited for monitoring the local changes in the structure of the investigated tRNA variants and when used as a set of probes can give a more complete picture of changes that occur in transcripts as a result of a mutation. PMID- 8718863 TI - Dissection of the ability of the chicken lysozyme gene 5' matrix attachment region to stimulate transgene expression and to dampen position effects. AB - The chicken lysozyme gene domain is flanked by nuclear matrix attachment regions (MARs) on each side. We have previously shown that bilaterally flanking 5'MARs in stably transfected artificial genetic units enhance expression of a reporter transgene and dampen position effects of the chromatin structure at the site of integration. The 5' MAR was now dissected into smaller fragments that were monitored for effects on transgene expression in mouse 3T3 cells by a similar assay. Fragments, which contain 1.32 and 1.45 kb and represent the upstream and the downstream half, respectively, of the 5' MAR, retained the ability to stimulate transgene expression as well as the ability to reduce the variation in the level of expression. However, a 452 bp subfragment (H1-HaeII), which still exhibits specific binding to nuclear matrices and contains two high-affinity binding sites for the abundant nuclear matrix protein ARBP, lost both of those abilities. A dimerized 177 bp sequence from fragment H1-HaeII, which also binds selectively to nuclear matrices and includes a duplicated ARBP binding site, was also unable to stimulate reporter gene expression. Furthermore, a 0.65 kb subfragment containing an intrinsically bent sequence did not affect an elevated reporter gene expression and its dampening. Our results show that the ability of MAR fragments to bind to nuclear matrices is not sufficient to enhance and insulate transgene expression in stably transfected cells. PMID- 8718864 TI - Substitution of serine for glycine-91 in the HXGH motif of CTP:phosphocholine cytidylyltransferase implicates this motif in CTP binding. AB - The effect of mutations in the proposed catalytic domain of CTP:phosphocholine cytidylyltransferase was investigated by constructing the single mutants CT-S91 and CT-C114 from the double mutant CT-S91C114, previously shown to have 4-fold lower than wild-type activity [Walkey, C.R., Kalmar, G. B., & Cornell, R. B. (1994) J. Biol. Chem. 269, 5742-5749]. The constructs were overexpressed in COS cells. The mutation Gly-91 to Ser-91 was found to be responsible for the decreased activity, whereas Ser-114 to Cys-114 had no effect. An alanine substitution at position 91, CT-A91, had a lesser effect on cytidylyltransferase activity. CT-S91 and CT-WT were purified from COS cells, and their kinetic constants were determined. CT-S91 had a 4-fold lower Vmax, and a K(m) for CTP 25 fold higher than the wild-type enzyme, suggesting that substitution of Gly-91 with serine interferes with CTP binding. The K(m) for phosphocholine was not affected in the CT-S91 mutant. There was no difference in the chymotrypsin sensitivities of CT-S91 and CT-WT, indicating that the mutation did not cause a global change in protein structure. However, the CT-S91 activity was more susceptible to inhibition by the denaturant urea than that of CT-WT, indicative of a perturbation of the active site folding. Gly-91 resides in the local sequence HSGH, which has been proposed to be a CTP-binding motif in the novel cytidylyltransferase superfamily [Bork, P., Holm, L., Koonin, E.V., & Sander, C. (1995) Proteins: Struct., Funct., Genet. 22, 259-266]. Our results represent the first experimental validation of this hypothesis. PMID- 8718865 TI - Cys102 and His398 are required for bleomycin-inactivating activity but not for hexamer formation of yeast bleomycin hydrolase. AB - The bleomycin-inactivating enzyme, bleomycin hydrolase, is believed to be involved in tumor resistance to the anticancer drug bleomycin. This homohexamer is an aminopeptidase that shows homology to cysteine proteinases around the cysteine and histidine active site. The role that these residues play in hydrolyzing bleomycin and in hexamer oligomerization of bleomycin hydrolase is not known. In this study, the yeast bleomycin hydrolase gene was expressed in Escherichia coli, and site-directed mutagenesis was employed to precisely investigate the roles of the conserved Cys102 and His398 residues in its structure and enzymatic activity. Three mutants were created, in which Cys102 was replaced by arginine or serine, and His398 was changed to glycine. The ability of bleomycin hydrolase to oligomerize was neither affected by the subtle cysteine/serine mutation nor affected by cysteine/arginine or histidine/glycine mutations. However, the ability of bleomycin hydrolase to hydrolyze and inactivate bleomycin was totally abolished in all three mutants, suggesting that the cysteine thiol and histidine imidazole are critical for hydrolyzing bleomycin. Furthermore, in contrast to predictions from the recently reported crystal structure of this enzyme, hexamer formation is not required for the enzymatic activity of bleomycin hydrolase. Thus, these results demonstrate that Cys102 and His398 are required for bleomycin hydrolase activity but not hexamer formation, and that both monomer and hexamer are active forms of bleomycin hydrolase. PMID- 8718866 TI - Mitochondrial phosphate transport protein. replacements of glutamic, aspartic, and histidine residues affect transport and protein conformation and point to a coupled proton transport path. AB - The homodimeric mitochondrial phosphate transport protein (PTP), which has six transmembrane helices per subunit, catalyzes inorganic phosphate transport in an electroneutral and pH gradient-dependent manner across the inner membrane. We have replaced the Glu, Asp, and His residues of the yeast PTP to assess their role in the transport mechanism. Mutants with physiologically relevant transport activity were identified by their ability to rescue the PTP null mutant yeast from glycerol medium. Five residues appear critical for transport: His-32 in helix A, Glu-126 and -137 in helix C, and Asp-39 and -236 at the matrix ends of helices A and E. These mutant PTPs are expressed at near normal levels in yeast. This yeast PTP and the mutants were expressed in Escherichia coli as inclusion bodies, solubilized, purified, and reconstituted. Their transport activities correlate well with the physiological assays. None of the transport inactivating mutations appear to be due to major protein conformation changes as assayed by the efficiency of PTP incorporation into liposomes. Only the Glu95Gln (cytosolic helices B and C-connecting segment), Glu163Gln and Glu164Gln (matrix helices C and D-connecting segment), and Glu126Asp (helix C) show a near 70% decrease in liposome incorporation efficiency. In addition, mutations at either end of helix D increase phosphate transport 2-fold. We would like to suggest that Glu-126, His 32, and Glu-137 (similar to Asp-96, Lys-216, and Asp-85 of bacteriorhodopsin) form a proton cotransport pathway that is coupled in an as yet undefined manner (possibly via His-32) to a phosphate transport pathway, which may include helix D. PMID- 8718867 TI - Time-resolved electrochromism associated with the formation of quinone anions in the Rhodobacter sphaeroides R26 reaction center. AB - The bacterial photosynthetic reaction center contains bacteriochlorophyll (Bchl) and bacteriopheophytin (Bph) cofactors that provide natural probes of electrostatic fields within this protein. We have examined the electrochromic responses of these cofactors, resolved during the lifetimes of the quinone anion states, P+QA-QB and P+QAQB-, and measured as a function of temperature. These measurements provide information on the time-dependent variation in electrostatic field strength on the Bchl and Bph cofactors. Measurements in the near-infrared absorbance bands have revealed the following. First, the QA-QB-->QAQB- electron transfer rate is found to be heterogeneous, consisting of at least two distinct kinetic components. At room temperature, we find a previously unresolved fast kinetic component with a reaction time of 25-40 microseconds, depending upon the preparation, that accounts for approximately 25% of the total reaction yield. The major component was identified with a reaction time of 210-240 microseconds. Below -20 degrees C, QA-QB-->QAQB- electron transfer shows distributed kinetics. The temperature-dependent conversion from biphasic to distributed kinetics suggests that there is a thermal averaging of conformational substates around two reaction center configurations. Interestingly, direct excitation of the Bph with 532 nm light at low temperatures appears to alter the electron transfer kinetics, possibly by inducing a change in the distribution of conformational states. The reaction kinetics were found to be sensitive to the addition of ethylene glycol, which is likely to reflect an osmolarity effect. Second, time-dependent absorption changes of the Bchl and Bph cofactors are found to be kinetically decoupled. The rapid responses of the Bph bands are interpreted to reflect electron transfer, while the slower responses of the Bchl are interpreted to reflect slower relaxation events, possibly including proton uptake. Finally, we find that the electrochromic response and QA-QB-->QAQB- electron transfer to be sensitive to the preparative state of the reaction center, reflecting differences in quinone binding for reaction centers in different states of purification. PMID- 8718868 TI - Polar residues in helix VIII of subunit I of cytochrome c oxidase influence the activity and the structure of the active site. AB - The aa3-type cytochrome c oxidase from Rhodobacter sphaeroides is closely related to eukaryotic cytochrome c oxidases. Analysis of site-directed mutants identified the ligands of heme a, heme a3, and CuB [Hosler et al. (1993) J. Bioenerg. Biomembr. 25, 121-133], which have been confirmed by high-resolution structures of homologous oxidases [Iwata et al. (1995) Nature 376, 660; Tsukihara et al. (1995) Science 269, 1069; (1996) 272, 1136]. Since the protons used to form water originate from the inner side of the membrane, and the heme a3-CuB center is located near the outer surface, the protein must convey these substrate protons to the oxygen reduction site. Transmembrane helix VIII in subunit I is close to this site and contains several conserved polar residues that could function in a rate-determining proton relay system. To test this role, apolar residues were substituted for T352, T359, and K362 in helix VIII and the mutants were characterized in terms of activity and structure. Mutation of T352, near CuB, strongly decreases enzyme activity and disrupts the spectral properties of the heme a3-CuB center. Mutation of T359, below heme a3, substantially reduces oxidase activity with only minor effects on metal center structure. Two mutations of K362, approximately 15 A below the axial ligand of heme a3, are inactive, make heme a3 difficult to reduce, and cause changes in the resonance Raman signal specific for the iron-histidine bond to heme a3. The results are consistent with a key role for T352, T359, and K362 in oxidase activity and with the involvement of T359 and K362 in proton transfer through a relay system now plausibly identified in the crystal structure. However, the characteristics of the K362 mutants raise some questions about the assignment of this as the substrate proton channel. PMID- 8718869 TI - Mechanistic and structural contributions of critical surface and internal residues to cytochrome c electron transfer reactivity. AB - The influence of mutations in two conserved regions of yeast iso-1-cytochrome c believed to be critical to the mechanism of cytochrome c electron transfer reactions has been investigated. The variants Asn52Ala, Tyr67Phe, Ile75Met, and Thr78Gly involve perturbation of critical hydrogen-bonding interactions with an internal water molecule (Wat166) and have been studied in terms of their electrochemical properties and the kinetics with which they are reduced by Fe(EDTA)2- and oxidized by Co(phen)3(3+). In parallel studies, the Co(phen)3(3+) oxidation kinetics of Tyr, Leu, Ile, Ala, Ser, and Gly variants of the phylogenetically conserved residue Phe82 have been studied and correlated with previous electrochemical and kinetic results. To assist mechanistic interpretation of these results, the three-dimensional structures of the Asn52Ala and Ile75Met ferrocytochrome c variants have been determined. The reduction potentials of the variants modified in the region of Wat166 were at least 33 mV (pH 6, 25 degrees C, and mu = 0.1 M) lower than that of the wild-type protein. Electron transfer reactivity of this family of variants in both the oxidation and reduction reactions was increased as much as 10-fold over that of the wild-type cytochrome. On the other hand, the reactivity of the position-82 variants in both oxidation and reduction depended on the structural characteristics of the oxidation-reduction reagent with which they reacted, and this reactivity was related to the nature of the residue at this position. These findings have been interpreted as demonstrating that the principal influence of modification at position-82 arises from changes in the nature of reactant-protein interaction at the surface of the protein and in maintaining the high reduction potential of the cytochrome while the principal influence of internal modifications near Wat166 results from alteration of the reorganization energy for the oxidation state linked conformational change defined by crystallographic analysis of the wild type protein. PMID- 8718870 TI - Fourier-transform infrared spectroscopy of phytochrome: difference spectra of the intermediates of the photoreactions. AB - The photocycle of 124 kDa phytochrome A from Avena sativa was studied by Fourier transform infrared spectroscopy at low temperatures. Difference spectra between the parent state Pr and the intermediates of the Pr-->Pfr pathway, i.e. lumi-R, meta-Ra, and meta-Rc, and between Pfr and the intermediates of the Pfr-->Pr pathway, lumi-F and meta-F, were obtained in 1H2O and 2H2O for the first time. Each spectrum shows characteristic spectral features which allow a clear distinction between the different intermediates. A general feature is that greater changes occur with increasing temperature, i.e. at the later steps of the photoreactions. Nevertheless, the changes in the spectral regions of the protein (amide I and amide II) were found to be surprisingly small, excluding larger conformational changes of the protein. All spectra of the intermediates are characterized by a strong negative band around 1700 cm-1. This band is tentatively assigned to the C = O stretch of ring D of the chromophore. Since it is not observed in the difference spectra between the parent states, it is concluded that ring D is located in a similar molecular environment in Pr and Pfr. In the photoproducts lumi-R and lumi-F, this band undergoes an upshift to 1720 cm-1. The high frequencies suggest that the chromophore is protonated in these intermediates as well as in Pr and Pfr. PMID- 8718871 TI - Factors affecting the H+/e- stoichiometry in mitochondrial cytochrome c oxidase: influence of the rate of electron flow and transmembrane delta pH. AB - A study is presented of the factors affecting the H+/e- stoichiometry of the proton pump of mitochondrial cytochrome c oxidase, isolated and reconstituted in phospholipid vesicles (COV). Under level flow conditions, i.e., in the absence of a transmembrane delta muH+, the H+/e- ratio, obtained from spectrophotometric measurements of the initial rates of electron flow and H+ release specifically elicited by cytochrome c, varied from around 0 to 1, depending on the actual rate of electron flow through the oxidase. At steady state the H+/e- ratio for the oxidase was specifically depressed by the transmembrane delta pH. The study of the H+/e- ratio of the pump was complemented by an analysis of the redox pattern of cytochrome c, CuA, and heme a. From both sets of results and recent structural data from other groups, it is concluded that the dependence of the H+/e- ratio on the rate of electron flow through the oxidase and transmembrane delta pH is associated with the possible occurrence of two electron transfer pathways in cytochrome c oxidase, a coupled one (cyt c-->CuA-->heme a-->heme a3-CuB) and a decoupled one (cyt c-->CuA-->heme a3-CuB). The contributions of the two pathways, differently affected by kinetics and thermodynamic factors, will determine the actual H+/e- ratio of the pump. A possible role of heme a in the proton pump and the physiological implication of the variable H+/e- ratio in the oxidase are discussed. PMID- 8718872 TI - Steric interaction between the 9-methyl group of the retinal and tryptophan 182 controls 13-cis to all-trans reisomerization and proton uptake in the bacteriorhodopsin photocycle. AB - The hypothesis was tested whether in bacteriorhodopsin (BR) the reduction of the steric interaction between the 9-methyl group of the chromophore all-trans retinal and the tryptophan at position 182 causes the same changes as observed in the photocycle of 9-demethyl-BR. For this, the photocycle of the mutant W182F was investigated by time-resolved UV-vis and pH measurements and by static and time resolved FT-IR difference spectroscopy. We found that the second half of the photocycle was similarly distorted in the two modified systems: based on the amide-I band, the protonation state of D96, and the kinetics of proton uptake, four N intermediates could be identified, the last one having a lifetime of several seconds; no O intermediate could be detected; the proton uptake showed a pronounced biphasic time course; and the pKa of group(s) on the cytoplasmic side in N was reduced from 11 in wild type BR to around 7.5. In contrast to 9-demethyl BR, in the W182F mutant the first part of the photocycle does not drastically deviate from that of wild type BR. The results demonstrate the importance of the steric interaction between W182 and the 9-methyl group of the retinal in providing tight coupling between chromophore isomerization and the late proton transfer steps. PMID- 8718873 TI - Structural distortions induced by integration host factor (IHF) at the H' site of phage lambda probed by (+)-CC-1065, pluramycin, and KMnO4 and by DNA cyclization studies. AB - Integration Host Factor (IHF) is a sequence-specific DNA-bending protein that is proposed to interact with DNA primarily through the minor groove. We have used various chemical probes [(+)-CC-1065, a minor-groove-specific agent that alkylates N3 of adenine and traps bends into the minor groove; pluramycin, a minor-major-groove threading intercalator that alkylates N7 of guanine; KMnO4, which reacts more strongly with bases in denatured DNA] to gain more information on the interaction of IHF with the H' site of phage lambda. In addition to the 13 bp core consensus recognition element present at all IHF binding sites, the H' site also has an upstream AT-rich element that increases the affinity of IHF for this site. Our results reveal new details of IHF-DNA interaction at this site. Results with (+)-CC-1065 modification suggest that IHF interacts with the adenines on the 3'-side of the AT-rich element and likely induces a minor-groove bend in its vicinity, which in turn stabilizes the interaction. Pluramycin modification experiments suggest the presence of both short- and long-range structural perturbations (possible DNA unwinding events) on either side of the IHF contact region. Although IHF is known to induce a large bend in DNA at the H' site, no separation of base pairs was detected when the bent DNA was probed with KMnO4. DNA cyclization studies indicate a large magnitude (approximately 180 degrees) for the IHF-induced bend at the H' site, consistent with > 140 degrees bend estimated by gel electrophoresis methods. These studies suggest that IHF induced DNA bending is accompanied by the introduction of a DNA node, DNA unwinding, and/or by some other DNA distortion. An enhanced binding and stability of IHF was observed on small circular DNA. PMID- 8718874 TI - Interaction of the periplasmic dG-selective Streptomyces antibioticus nuclease with oligodeoxynucleotide substrates. AB - The interaction of a periplasmic nuclease, isolated from Streptomyces antibioticus, with several oligodeoxynucleotide substrates has been studied. Double-stranded oligonucleotides that contain sequences of four or more consecutive deoxyguanosine residues are preferentially hydrolyzed, with the strongest cutting site occurring at GGG decreases G. The enzyme does not hydrolyze these sequences in single-stranded DNA. However the sequence selectivity of the nuclease is far from absolute. Other sequences can also be cut, albeit more poorly, and differences in cutting rates are observed for runs of dG bases that differ in their flanking sequences. An oligonucleotide, thirty six bases in length, that contains a central run of five dG bases has been used to evaluate the importance of the individual deoxyguanosines in recognition and cleavage. With this oligonucleotide cutting takes place at GG[symbol: see text]G decreases G[symbol: see text]G (decreases, most prominent cut; [symbol: see text], less prominent cuts). The use of dG base analogues revealed that two bases, one and two steps removed from the cleavage site in the 5' direction (*G*GG decreases), were of most importance in the determination of the nuclease DNA cleavage selectivity. Of these the inner starred dG was the most critical. The use of 5-methyldeoxycytidine also showed that the dC, base paired to this critical dG, influenced cleavage specificity. The overall pattern of results seen with the base analogues suggested that the nuclease interacted with both strands of the DNA and also contacted the nucleic acid in both the major and minor grooves. Gel retardation analysis together with footprinting experiments using hydroxyl radicals, dimethyl sulfate, and ethylnitrosourea indicated that the nuclease does not form a tight and specific complex with sequences containing dG runs, at least in the absence of the essential co-factor, Mg2+. PMID- 8718875 TI - The stability of abortively cycling T7 RNA polymerase complexes depends upon template conformation. AB - We have developed a promoter competition assay to determine whether T7 RNA polymerase dissociates from its template during abortive cycling. We find that the stability of the initiation complex (IC) depends upon the conformation of the promoter, and that the degree to which the template is unwound contributes importantly to the stability of the IC. On linear DNA or a relaxed plasmid template, the stability of the IC is very low (t1/2 < 1 min). However, on a supercoiled template, the IC has a stability that is comparable to that of a paused elongation complex (t1/2 = 14 min). At a synthetic promoter that is single stranded in the initiation region (from -5 and downstream), the polymerase forms a highly stable complex (t1/2 > 30 min) even in the absence of RNA synthesis. These findings are important to our understanding of the transition from the IC to an EC. PMID- 8718876 TI - Conformational and functional study of magainin 2 in model membrane environments using the new approach of systematic double-D-amino acid replacement. AB - Systematic double-D-amino acid replacement of adjacent amino acids has been used to study the secondary structure of the amphiphilic, antibiotic peptide magainin 2 amide (M2a) by circular dichroism spectroscopy. Bound to liposomes, the secondary structure of the peptide is characterized by a weak alpha-helix in the N-terminus and a stable alpha-helix between residues 9 and 21. The lack of conformational differences in the peptide when bound to vesicles of varying negative charge density indicates marked independence of the structure from electrostatic forces. The similarity of the helicity profiles observed for double D-isomers bound to vesicles and in the presence of sodium dodecyl sulfate micelles (SDS) clearly shows that SDS can mimic magainin-lipid interactions. In contrast, in 1:1 trifluoroethanol/buffer (v/v), the peptide exhibits a weak alpha helix extended from the N- to the C-terminus. Dye release experiments from vesicles of phosphatidylglycerol showed that double-D-amino acid substitution only in the region of the stable helix results in a reduction of the membrane permeabilizing ability. On vesicles with a reduced amount of acidic phospholipids, double-D-amino acid substitution in any position leads to a drastic reduction of peptide-induced membrane permeabilization. Whereas the activity of M2a on phosphatidylglycerol was found to be mainly electrostatically determined, hydrophobic interactions play a decisive role in the interaction with vesicles of reduced negative charge density. Fluorescence investigations of tryptophan-containing analogs of high and low helicity showed that differences in the location of the chromophores of the membrane-bound peptides do not exist. PMID- 8718877 TI - Biochemical evaluation of photolabile precursors of choline and of carbamylcholine for potential time-resolved crystallographic studies on cholinesterases. AB - Acetylcholinesterase and butyrylcholinesterase both rapidly hydrolyze the neurotransmitter acetylcholine. The unusual three-dimensional structure of acetylcholinesterase, in which the active site is located at the bottom of a deep and narrow gorge, raises cogent questions concerning traffic of the substrate, acetylcholine, and the products, choline and acetate, to and from the active site. Time-resolved crystallography offers a promising experimental approach to investigate this issue but requires a suitable triggering mechanism to ensure efficient and synchronized initiation of the dynamic process being monitored. Here we characterize the properties of two photolabile triggers which may serve as tools in time-resolved crystallographic studies of the cholinesterases. These compounds are 2-nitrobenzyl derivatives of choline and of carbamylcholine, which generate choline and carbamylcholine, respectively, upon photochemical fragmentation. Both photolabile compounds are reversible inhibitors, which bind at the active sites of acetylcholinesterase and butyrylcholinesterase with inhibition constants in the micromolar range, and both photofragmentation processes occur rapidly and with a high quantum yield, without substantial photochemical damage to the enzymes. Photolysis both of acetylcholinesterase and of butyrylcholinesterase, complexed with a 2-nitrobenzyl derivative of choline, resulted in regeneration of enzymic activity. Photolysis of acetylcholinesterase complexed with the 2-nitrobenzyl derivative of carbamylcholine led to time dependent inactivation, resulting from carbamylation of acetylcholinesterase, which could be reversed upon dilution, due to decarbamylation. Both sets of experiments demonstrated release of choline within the active site. In the former case, choline was produced photochemically at the active site. In the latter case, choline was generated enzymatically, within the active site, concomitantly with carbamylation of the acetylcholinesterase. The two photolabile compounds may thus serve as complementary probes for time-resolved studies of the route of product release from the active sites of the cholinesterases. PMID- 8718878 TI - The A and B tubules of the outer doublets of sea urchin sperm axonemes are composed of different tubulin variants. AB - The alpha beta-tubulin heterodimer, the structural unit of microtubules, comes in many variants. There are different alpha and beta isotypes encoded by multigene families. Additional heterogeneity is generated by a set of posttranslational modifications. Detyrosination of alpha-tubulin, removal of the carboxy-terminal Glu-Tyr dipeptide of alpha-tubulin, phosphorylation of some tubulins, polyglutamylation, and polyglycylation of alpha- and beta-tubulins all involve the acidic carboxy-terminal region. We have investigated the distribution of tubulin variants in the axonemal microtubules of sea urchin sperm flagella by immunological procedures and by direct sequence and mass spectrometric analysis of the carboxy-terminal peptides. The A and B tubules that comprise the nine outer doublets differ strongly in tubulin variants. A tubules contain over 95% unmodified, tyrosinated alpha beta-tubulin. In B tubules, alpha-tubulin is approximately 65% detyrosinated and both alpha- and beta-tubulin are 40-45% polyglycylated. These results show a segregation of tubulin variants between two different axonemal structures and raise the possibility that posttranslational modifications of tubulins reflect or specify structurally and functionally distinct microtubules. PMID- 8718879 TI - Biogenesis and topology of integral membrane proteins: characterization of lactose permease-chloramphenicol acetyltransferase hybrids. AB - Use of beta-lactamase in gene fusions to study membrane protein topology permits exploitation of its biological activity to select for positive (external) hybrids on ampicillin agar plates. When the enzyme is attached to cytoplasmic loops of a membrane protein, it is not secreted and is therefore unable to confer ampicillin resistance. In this study, we examine the use of the cytoplasmic enzyme chloramphenicol acetyltransferase (Cat) as a complement to the use of periplasmic beta-lactamase, in gene fusion studies. This enzyme is responsible for chloramphenicol resistance in Escherichia coli. We show that Cat confers substantial antibiotic resistance when fused to cytoplasmic loops of lactose permease. As expected, periplasmically exposed Cat is enzymatically active in vitro but unable to confer significant chloramphenicol resistance, presumably because of the absence of acetylcoenzyme A in the periplasm. Therefore, Cat may serve as a topogenic sensor in gene fusion studies. The new Cat fusion approach is discussed with regard to its potential use for selecting E. coli mutants which are defective in the assembly of membrane proteins. PMID- 8718880 TI - Identification of active site residues essential to 4-chlorobenzoyl-coenzyme A dehalogenase catalysis by chemical modification and site directed mutagenesis. AB - 4-Chlorobenzoyl-coenzyme A (4-CBA-CoA) dehalogenase catalyzes the hydrolysis of 4 CBA-CoA to 4-hydroxybenzoyl-coenzyme A (4-HBA-CoA) via a nucleophilic aromatic substitution pathway involving the participation of an active site carboxylate side chain in covalent catalysis. In this paper we report on the identification of conserved aspartate, histidine, and tryptophan residues essential to 4-CBA-CoA catalysis using chemical modification and site-directed mutagenesis techniques. Treatment of the dehalogenase with diethyl pyrocarbonate resulted in complete loss of catalytic activity (Kinact = 0.17 mM-1 min-1 at pH 6.5, 25 degrees C) that was fully regained by subsequent treatment with hydroxylamine. The protection from inactivation afforded by enzyme bound 4-HBA-CoA indicated that the essential histidine residues are located at the active site. Replacement of conserved histidine residues 81, 90, 94, and 208 with glutamine residues resulted in a significant loss of catalytic activity only in the cases of the histidine 81 and 90 mutants. Substrate and product ligand binding studies showed that binding is not significantly inhibited in these mutants. Site directed mutagenesis of a selection of conserved aspartate and glutamate residues, identified aspartate 145 as being essential to dehalogenase catalysis. Ligand binding studies showed that this residue is not required for tight substrate/product binding. Chemical modification of the dehalogenase with N-bromosuccinimide resulted in full loss of catalytic activity that was prevented by saturation of the active site with product ligand, providing evidence favoring an essential active site tryptophan. Phenylalanine replacement of conserved tryptophan residues 179 and 137 reduced catalytic activity only in the latter (Kcat = 0.03% of wild-type dehalogenase). On the basis of these results and the recently determined X-ray crystal structure of the complex of 4-CBA-CoA dehalogenase and 4-HBA-CoA [Benning, M. M., Taylor, K.L., Liu, R.-Q., Yang, G., Xiang, H., Wesenberg, G., Dunaway-Mariano, D., Holden, H.M. (1996) Biochemistry 35,8103-8109] we propose that aspartate 145 functions as the active site nucleophile, that tryptophan 137 serves as a hydrogen bond donor to the aspartate 145 C = O, and that histidine 90 serves to deprotonate the bound H2O molecule. PMID- 8718881 TI - Differential effects of ubiquitin aldehyde on ubiquitin and ATP-dependent protein degradation. AB - ATP-dependent proteolysis of 125I-labeled human alpha-globin, bovine alpha lactalbumin, bovine serum albumin, or chicken lysozyme was assessed in a rabbit reticulocyte extract supplemented with ATP, excess ubiquitin, and variable amounts of ubiquitin aldehyde (Ubal), an inhibitor of many ubiquitin-protein isopeptidases. Low concentrations (0.8 microM) of Ubal increased the ATP dependent degradation of 125I-alpha-globin by approximately 30% after 2 h at 37 degrees C, had little effect on 125I-lysozyme turnover, and decreased 125I-alpha lactalbumin or 125I-albumin degradation by approximately 20%. The ATP-dependent degradation of all substrates was inhibited by high concentrations (> 3 microM) of Ubal throughout the incubation (15 min to 2 h); after 2 h, this inhibition ranged from 15% for 125I-alpha-globin to approximately 85% for 125I-alpha lactalbumin and 125I-albumin. Levels of ubiquitin-125I-protein conjugates were increased significantly with Ubal; with > or = 8.0 microM Ubal, high molecular mass multiubiquitinated conjugates were particularly evident for 125I-alpha globin and 125I-alpha-lactalbumin. These mixtures also accumulated ubiquitin conjugates with sizes expected for di- through pentaubiquitin oligomers. The results are consistent with the following proposed events: The ATP-dependent degradation of 125I-alpha-lactalbumin or 125I-albumin is probably mediated almost exclusively through polyubiquitinated intermediates. High Ubal concentrations inhibit an isopeptidase(s) which normally disassembles "unanchored" polyubiquitin chains that remain after substrate degradation by the 26S proteasome; these chains accumulate to inhibit further conjugate degradation. Much of the ATP dependent degradation of 125I-alpha-globin and, to a lesser degree, 125I-lysozyme may occur through alternative structures where ubiquitin monomers or short oligomers are ligated to one or more substrate lysines. For 125I-alpha-globin, even low concentrations of Ubal effectively inhibit deubiquitination of these conjugates to enhance alpha-globin degradation. PMID- 8718882 TI - Identification of functional domains within the alpha and beta subunits of beta hexosaminidase A through the expression of alpha-beta fusion proteins. AB - There are three human beta-hexosaminidase isozymes which are composed of all possible dimeric combinations of an alpha and/or a beta subunit; A (alpha beta), and B (beta beta), and S (alpha alpha). The amino acid sequences of the two subunits are 60% identical. The homology between the two chains varies with the middle > the carboxy-terminal > > the amino-terminal portions. Although dimerization is required for activity, each subunit contains its own active site and differs in its substrate specificity and thermal stability. The presence of the beta subunit in hexosaminidase A also influences the substrate specificity of the alpha subunit; e.g., in vivo only the A heterodimer can hydrolyze GM2 ganglioside. In this report, we localize functional regions in the two subunits by cellular expression of alpha/beta fusion proteins joined at adjacently aligned residues. First, a chimeric alpha/beta chain was made by replacing the least well conserved amino-terminal section of the beta chain with the corresponding alpha section. The biochemical characteristics of this protein were nearly identical to hexosaminidase B. Therefore, the most dissimilar regions in the subunits are not responsible for their dissimilar biochemical properties. A second fusion protein was made that also included the more homologous middle section of the alpha chain. This protein expressed the substrate specificity unique to isozymes containing an alpha subunit (A and S). We conclude that the region responsible for the ability of the alpha subunit to bind negatively charged substrates is located within residues alpha 132-283. Interestingly, the remaining carboxy terminal section from the beta chain, beta 316-556, was sufficient to allow this chimera to hydrolyze GM2 ganglioside with 10% the specific activity of heterodimeric hexosaminidase A. Thus, the carboxy-terminal section of each subunit is likely involved in subunit-subunit interactions. PMID- 8718883 TI - Metal-substrate interactions facilitate the catalytic activity of the bacterial phosphotriesterase. AB - The bacterial phosphotriesterase from Pseudomonas diminuta is a zinc metalloenzyme which catalyzes the hydrolysis of a variety of organophosphorus nerve agents with high efficiency. The active site of the enzyme consists of a coupled binuclear metal center embedded within a cluster of histidine residues. Potential protein-substrate interactions at the active site were probed by a systematic variation of metal identity, leaving group potential, phosphate host, and amino acid replacement. In order to determine the roles of these metal ions in binding and catalysis, the microscopic rate constants and kinetic parameters were obtained with various divalent cations. The divalent cations that were utilized in this investigation consisted of Co2+, Ni2+, Cd2+, Zn2+, Mn2+, and the mixed-metal Zn2+/Cd2+ hybrid. The leaving group potential and phosphate host were varied by altering the pKa of the departing substituted phenol or thiophenol in either a diethyl phosphate or a diethyl thiophosphate substrate. The Bronsted plots for the nonenzymatic hydroxide catalyzed hydrolysis of these substrates showed a linear dependence between the pseudo-first-order rate constant and the pKa of the leaving group. Enzymatic activities of the wild-type enzyme with these same substrates varied by over 7 orders of magnitude over the entire experimental pKa range (4.1-10.3), and the corresponding Bronsted plots were nonlinear. Those substrates with leaving groups with high pKa values were limited by the rate of bond cleavage while those substrates having leaving groups with low pKa values were limited by a conformational change or binding event. Thiophosphate substrates having leaving groups with high pKa values were better substrates than the corresponding phosphate analogues. These results are consistent with the direct coordination of one or both metal ions with the phosphoryl sulfur or oxygen atom of the substrate. A large dependence of the rate on the leaving group rules out the possibility of protonation of the leaving group or electrostatic interaction of the leaving group oxygen (or sulfur) with a metal ion or cationic group at the active site. The large differences in the size of the beta lg over the range of metal ions utilized by the enzyme indicate that the metal ions polarize the phosphoryl group and alter the structure of the transition state. The values of V/K(m) for the enzyme-catalyzed hydrolysis for a series of substituted thiophenol analogues were 10(2)-10(3)-fold smaller than those obtained for the hydrolysis of the corresponding phenolic substrates, suggesting that the bulkier sulfur substituent in the leaving group may induce conformational restrictions at the active site. With the zinc-substituted H201N mutant enzyme, there was a large decrease in the rate of phosphotriester hydrolysis but essentially no change in the rate of thiophosphotriester hydrolysis relative to the values observed for the zinc-substituted wild-type enzyme. These results suggest that a direct perturbation in the ligand structure of the binuclear metal center induces alterations in the mechanism of substrate hydrolysis. PMID- 8718885 TI - Kinetics of the phosphorylation of Na,K-ATPase by inorganic phosphate detected by a fluorescence method. AB - Phosphorylation by Pi of the Na,K-ATPase from rabbit kidney in the absence of Na+ ions but in the presence of Mg2+ ions has been studied. In the absence of K+ ions, unphosphorylated and phosphorylated states induce different fluorescence levels in the membrane-bound styryl dye RH421, and hence transitions between the two states were monitored. Transient kinetic studies of phosphorylation were initiated by manual addition of Pi or by photochemical release of Pi from 1-(2 nitrophenyl)ethyl phosphate (caged Pi) using laser flash photolysis at 308 nm. Equilibrium studies of phosphorylation showed that the apparent Km for Pi was 23.0 +/- 0.3 microM (mean +/- sem) at pH 7.1 and 21 degrees C. The dye fluorescence increased in a biphasic manner on addition of 500 microM Pi to the enzyme: a rapid phase (t 1/2 < 1 s) and a slower exponential phase at 0.059 +/- 0.003 s-1. The rate of the rapid phase was studied by fast concentration-jump experiments and exhibited first-order kinetics in Pi up to 60 microM. Fluorescence records vs time were exponential, and a plot of the rate constant versus [Pi] had a slope of 1.47 x 10(5) M-1 s-1 and ordinate [Pi] = 0) intercept of 3.1 s-1. Addition of 50 mM NaCl to the phosphorylated enzyme induced an exponential decay in the dye fluorescence from which a rate constant of 0.10 +/- 0.005 s-1 was determined. These data were interpreted in terms of transformations between conformational states E1 and E2, and the phosphorylated state P-E2 defined in the Post-Albers mechanism of the Na,K-ATPase [Lauger, P., (1991) Electrogenic Ion Pumps, Sinauer Associates Inc., Sunderland, MA] as follows: [formula: see text] The RH421 fluorescence of state P-E2 was studied over the pH range 6-8.5. Fluorescence was greatest at pH 8.5 and lowest at pH 6.0 in a simple binding isotherm with pK 7.5. The apparent Km for Pi rose cooperatively with increasing pH (pKa 8.6 and a Hill coefficient of 2). Therefore in the absence of monovalent metal ions, occupation of the cation (K+) binding sites by protons promotes phosphorylation by Pi. PMID- 8718884 TI - Proteolysis regulates exposure of the IIICS-1 adhesive sequence in plasma fibronectin. AB - The alternatively spliced type III connecting segment (IIICS) of fibronectin (Fn) contains an amino acid sequence, CS-1, which is recognized by the integrin receptor, alpha 4 beta 1. Plasma Fn inhibits alpha 4 beta 1-dependent binding of lymphocytes and monocytes to CS-1 containing Fn derivatives poorly, suggesting limited exposure of the CS-1 sequence in Fn. To test the availability of CS-1 in plasma Fn, an antibody was raised to the synthetic peptide CS-1. The CS-1 sequence was found to be minimally exposed in plasma Fn; and immobilization of Fn, a model of matrix deposition, caused only a modest increase in its exposure. Digestion of Fn with selected proteases, however, induced substantial expression of the CS-1 sequence. The acid protease cathepsin D generated fragments of 31 33.5 kDa from the COOH-terminal heparin-binding domain of Fn which possessed high immunoreactivity with anti-CS-1. Digestion of Fn with cathepsin B also resulted in the exposure of CS-1 sequence in a 140 kDa fragment. Although the digestion of Fn with neutral proteases (neutrophil elastase, cathepsin G, chymotrypsin, trypsin) generated fragments from the COOH-terminal heparin-binding domain of similar molecular weight as with cathepsin D, the exposure of CS-1 did not occur. Exposure of the CS-1 region by the cathepsins was supported by cell adhesion experiments; digestion of Fn with cathepsins D and B transformed inert plasma Fn to an effective inhibitor of adhesion of lymphoblastoid B and T cells (Ramos, Jurkat, Molt-4) to an immobilized CS-1 conjugate. These results suggest that exposure of the CS-1 sequence in plasma Fn by proteolysis with cathepsins D and B, enzymes implicated in several pathological processes, may serve a regulatory function in cell adhesion. The adhesive function of the CS-1 region in intact Fn appears to be suppressed by the native conformation of the molecule. PMID- 8718886 TI - Time resolution of binding and membrane insertion of a mitochondrial signal peptide: correlation with structural changes and evidence for cooperativity. AB - Utilizing a recently developed novel fluorescence technique [Wall et al. (1995) Mol. Membr. Biol. 12, 183-192], it is shown that the interactions of p25, the leader peptide of subunit IV of cytochrome c oxidase, with phospholipid membranes can be identified in real time. p25 is observed to bind following stopped-flow mixing of the peptide with phospholipid membranes with rate constants up to about 700 s-1 and then insert into the membrane with rate constants on the order of 0.4 s-1. Comparison of these processes with similarly time-resolved experiments performed with a stopped-flow CD spectrometer revealed that p25 does not become alpha-helical upon binding to the membrane. Following membrane insertion, however, p25 was observed to adopt an alpha-helical configuration. The temperature dependency of these processes was then found to yield activation energies for the respective components of the p25-membrane interaction. PMID- 8718887 TI - Mutational analysis of the catalytic subunit of the yeast vacuolar proton translocating ATPase. AB - In order to generate a set of tools for probing structure-function relationships in the catalytic subunit of the yeast vacuolar H(+)-ATPase, the gene encoding this subunit (VMA1) was randomly mutagenized. Mutant plasmids unable to complement the growth defects of yeast cells lacking an intact VMA1 gene were isolated and sequenced. Eight different mutant alleles of VMA1 were examined for levels of the catalytic subunit and other subunits of the enzyme, assembly of the ATPase complex, targeting to the vacuolar membrane, and concanamycin A-sensitive ATPase activity. The mutations S811P and E740D resulted in mutant enzymes that assembled fully but were incapable of ATP hydrolysis, and the mutation E785G generated a similar but somewhat less severe phenotype (17% of the ATPase activity of wild-type vacuoles). When MgATP-dependent stripping of the peripheral subunits by 100 mM KNO3 was examined in these three mutants, only the E785G mutant exhibited significant stripping, suggesting that ATP hydrolysis, even at relatively low levels, generates a conformation susceptible to dissociation. Plasmids containing the mutations E751G and F752S partially complemented the growth defects and resulted in partial defects in ATPase activity that appear to reflect reduced catalytic efficiency. Partial defects in growth and ATPase activity were also seen in the Y797H mutant, but this mutation caused an assembly defect manifested as a preferential loss of two of the peripheral subunits of the enzyme. The phenotypes of these mutants are interpreted in the context of homologies with other V-type and F-type ATPases. PMID- 8718888 TI - Insight into tyrosine phosphorylation in v-Fps using proton inventory techniques. AB - The phosphoryl group transfer step in the kinase domain of v-Fps, a nonreceptor tyrosine protein kinase, was analyzed using proton inventory and viscosometric techniques. The latter studies show that two nine-residue peptide substrates for v-Fps, peptides I (EAEAYEAIE) and II (EAEIYEAIE), are in rapid equilibrium with the active site and bind with similar affinities (Ks = 2.2 and 1.7 mM for peptides I and II). While phosphoryl group transfer is the rate-limiting step in kcat for peptide I (5 s-1) at neutral pH, peptide II is converted to product by a kinetic mechanism in which phosphoryl group transfer (45 s-1) and product release (20 s-1) partially control this parameter. Significant solvent isotope effects on kcat (k0/kn approximately 1.6) are observed for the phosphorylation of both peptides in 95% D2O. Proton inventories on kcat for peptide I are linear, indicating that the phosphoryl group transfer step is associated with a single proton transfer. Conversely, proton inventories on kcat for peptide II are "bowed" up, consistent with a "virtual" transition state in which phosphoryl group transfer and product release steps partially control this parameter. The lack of solvent isotope effects on kcat/Kpeptide for both peptides can be explained by an equilibrium isotope effect on substrate binding that offsets the kinetic isotope effect for phosphoryl group transfer. In keeping with this proposal, the KI for the inhibitor peptide, EAEIFEAIE, is 11 +/- 0.80 and 6.5 +/- 0.82 mM in 0 and 60% D2O, respectively. Fitting of the proton inventory plots using modified forms of the Gross-Butler equation provide intrinsic isotope effects of 1.7 and 3.6 for peptides I and II. The combined data are consistent with a mechanism involving either an acid-base catalyst or a conformational change preceding the release of products that is accompanied by the disruption of a single hydrogen donor-acceptor pair. PMID- 8718889 TI - The contribution of lysine-36 to catalysis by human myo-inositol monophosphatase. AB - The role of lysine residues in the catalytic mechanism of myo-inositol monophosphatase (EC 3.1.3.25) was investigated. The enzyme was completely inactivated by amidination with ethyl acetimidate or reductive methylation with formaldehyde and cyanoborohydride. Activity was retained when the active site was protected with Mg2+, Li+, and D,L-myo-inositol 1-phosphate. Using radiolabeling, peptide mapping, and sequence analysis, Lys-36 was shown to be the protected residue, which is responsible for inactivation. Replacing Lys-36 with glutamine produced a mutant protein, K36Q, with similar affinities for the substrate and the activator Mg2+, but a 50-fold lower turnover number as compared to the wild type enzyme. Crystallographic studies did not indicate any gross structural changes in the mutant as compared to the native form. Initial velocity data were best described by a rapid equilibrium ordered mechanism with two Mg2+ binding before and a third one binding after the substrate. Inhibition by calcium was unaffected by the mutation, but inhibition by lithium was greatly reduced and became noncompetitive. The pH dependence of catalysis and the solvent isotope effect on kcat are altered in the mutant enzyme. D,L-myo-Inositol 1-phosphate, 4 nitrophenyl phosphate, and D-glucose 6-phosphate are cleaved at different rates by the wild-type enzyme, but with similar efficiency by K36Q. All data taken together are consistent with the hypothesis that modifying or replacing the lysine residue in position 36 decreases its polarizing effect on one of the catalytic metal ions and prevents the efficient deprotonation of the metal-bound water nucleophile. PMID- 8718890 TI - 2.3 A resolution X-ray crystal structure of the bisubstrate analogue inhibitor salicylhydroxamic acid bound to human myeloperoxidase: a model for a prereaction complex with hydrogen peroxide. AB - The X-ray crystal structure of a salicylhydroxamic acid (SHA) inhibitory complex with human myeloperoxidase (MPO) has been determined at 2.3 A resolution. The aromatic ring of the inhibitor binds to a hydrophobic region at the entrance to the distal heme pocket between heme pyrrole ring D and the side chain of Arg 239. The hydroxamic acid moiety is hydrogen bonded to both the distal histidine 95 and the adjacent glutamine 91 amide group but is not coordinated to the heme iron. SHA binding displaces three water molecules from the distal heme cavity and causes a small shift in the position of a fourth water molecule. Otherwise, there are no significant conformational differences between the active site regions of the complex and the native enzyme. The ability of the three SHA oxygen atoms to closely duplicate the hydrogen-bonding pattern of these three water molecules in the native enzyme is postulated to account for the strong binding of this inhibitor to MPO. The mode of binding of SHA to MPO provides information on the binding sites for aromatic peracid substrates that promote compound I formation as well as aromatic alcohols and amines that carry out single-electron reductions of compound I. Similarities in the hydrogen-bonding patterns of amino acid residues and water molecules in the distal heme pockets of myeloperoxidase and the nonhomologous cytochrome c peroxidase suggest that they may have similar mechanisms of compound I formation. A model is presented for a prereaction complex of myeloperoxidase in which hydrogen peroxide is hydrogen bonded to the distal histidine, as a prerequisite for deprotonation and subsequent binding at the sixth coordination site of the heme iron. PMID- 8718891 TI - Inhibitor-induced changes in the intrinsic fluorescence of human cyclooxygenase 2. AB - The steady state tryptophan fluorescence of apo-human cyclooxygenase-2 (hCox-2) is quenched approximately 40%-50% by the slow binding inhibitors diclofenac, indomethacin, ketoprofen, NS-398, and DuP-697. The effects of these inhibitors on tryptophan fluorescence are both time and concentration dependent. Addition of each inhibitor results in a rapid fluorescence decrease, followed by a slower time dependent quenching. The slow, time dependent loss of fluorescence follows first-order kinetics, the rate constants for the process increasing with inhibitor concentration in a saturation-type manner. The rapid fluorescence loss also increases with increasing inhibitor concentration in the same manner. These results are consistent with the initial formation of a rapid equilibrium complex of enzyme and inhibitor (EI), followed by the slower formation of a tightly bound enzyme-inhibitor complex (EI*). The fluorescence of the EI complex is not significantly different from that of the EI* complex. The kinetic parameters of each inhibitor derived for this process (Ki and kon) are close to those obtained by determination of the rate constants for the onset of enzyme inhibition, thereby linking the fluorescence changes with inhibitor binding. The reversible inhibitors ibuprofen and docosahexaenoic acid do not quench the protein fluorescence but do decrease both the rate of the slow fluorescence loss and the magnitude of the initial rapid fluorescence decrease caused by the slow binding inhibitors, consistent with their competitive behavior. ASA-acetylated apo-hCox-2 shows the same fluorescence-quenching behavior in the presence of most of the above inhibitors. However, acetylation apparently blocks the binding of diclofenac, whereas the affinity of ibuprofen is increased. The effects of the collisional quenching agents iodide and acrylamide on both the native and inhibited enzyme are small (< 20% quenching at 0.3 M), showing that inhibitor binding does not result in an increased solvent accessibility of protein tryptophans. The cause of the inhibitor-induced quenching of the intrinsic apo hCox-2 fluorescence is likely energy transfer to the bound inhibitor. Calculations based on the inhibitor-tryptophan distances in ovine Cox-1 indicate that the distances are within the required range for significant quenching to occur. PMID- 8718892 TI - Calorimetric analysis of triple helices targeted to the d(G3A4G3).d(C3T4C3) duplex. AB - We present a thermodynamic analysis based on differential scanning calorimetry (DSC) of three short intermolecular DNA triplexes targeted to the same DNA duplex: d(C+3T4C+3)*d- (G3A4G3).d(C3T4C3) (PYR), d(G3A4G3)*d(G3A4G3).d(C3T4C3) (PUR), and d(G3T4G3)*d(G3A4G3).d(C3T4C3) (PUR/PYR). Enthalpies, delta H, and entropies, delta S, are measured by model-free integration of the DSC curves and are compared to the same quantities determined by van't Hoff analysis of the DSC curves and, in the case of the PYR and PUR/PYR triplexes, UV melting curves as well. In the case of the PUR triplex, which exhibits monophasic melting behavior, the calorimetric delta H and the calorimetrically determined van't Hoff delta H are in excellent agreement, indicating an all-or-none transition for this triplex. For the PYR and PUR/PYR triplexes, which melt in a biphasic manner, the calorimetrically determined van't Hoff delta H values are somewhat larger than the model-independent calorimetric delta H values. In those cases, however, good agreement is found between the calorimetric delta H values and the spectrophotometrically determined van't Hoff delta H values. The calorimetrically determined delta H values, expressed per mole of triplet, for the three triplexes are 4.5, 3.8, and 2.4 kcal/mol for the PUR, PYR, and PUR/PYR triplexes, respectively. The same order of stability is observed in terms of delta G and Tm values. The high stability of the PUR triplex at neutral pH indicates that purine oligonucleotides may be the most effective at targeting duplex regions for triple helix formation in vivo. PMID- 8718893 TI - Aspartate 74 as a primary determinant in acetylcholinesterase governing specificity to cationic organophosphonates. AB - Through site-specific mutagenesis, we examined the determinants on acetylcholinesterase which govern the specificity and reactivity of three classes of substrates: enantiomeric alkyl phosphonates, trifluoromethyl acetophenones, and carboxyl esters. By employing cationic and uncharged pairs of enantiomeric alkyl methylphosphonyl thioates of known absolute stereochemistry, we find that an aspartate residue near the gorge entrance (D74) is responsible for the enhanced reactivity of the cationic organophosphonates. Removal of the charge with the mutation D74N causes a near equal reduction in the reaction rate constants for the Rp and Sp enantiomers and exerts a greater influence on the cationic organophosphonates than on the charged trimethylammonio trifluoromethyl acetophenone and acetylthiocholine. This pattern of reactivity suggests that the orientation of the leaving group for both enantiomers is directed toward the gorge exit and in apposition to Asp 74. Replacement of tryptophan 86 with alanine in the choline subsite also diminishes the reaction rates for cationic organophosphonates, although to a lesser extent than with the D74N mutation, while not affecting the reactions with the uncharged compounds. Hence, reaction with cationic OPs depends to a lesser degree on Trp 86 than on Asp 74. Docking of Sp and Rp cycloheptyl methylphosphonyl thiocholines and thioethylates in AChE as models of the reversible complex and transition state using molecular dynamics affords structural insight into the spatial arrangement of the substituents surrounding phosphorus prior to and during reaction. The leaving group of the Rp and Sp enantiomers, regardless of charge, is directed to the gorge exit and toward Asp 74, an orientation unique to tetrahedral ligands. PMID- 8718894 TI - Identification of an osteoblastic silencer element in the first intron of the rat osteocalcin gene. AB - The osteocalcin gene has been used as a model for studying the regulation of gene expression by 1,25-dihydroxyvitamin D3, as well as for examining factors which contribute to osteoblast-specific regulation of gene expression. Most of these studies have focused on transactivation. We report the identification of a sequence in the first intron of the rat osteocalcin gene which suppresses the expression of osteocalcin-CAT fusion genes approximately 10-fold in ROS 17/2.8 and UMR 106 osteosarcoma cells. Mutation of a TTTCTTT motif in the first intron abolishes this suppression. The silencing effect of this motif is also observed after bone morphogenic protein-2 (BMP-2)-induced expression of the osteoblastic phenotype in the MLB13MYC clone 17 cell line. Mutation of the splice donor site does not affect suppression by these sequences in ROS 17/2.8 cells. When multimerized and placed upstream of the native osteocalcin promoter, these sequences retain their ability to mediate transcriptional repression. Electrophoresis mobility shift analysis demonstrates a specific protein-DNA interaction with the TTTCTTT motif in nuclear extracts from ROS 17/2.8, UMR 106, and MLB13MYC clone 17 cells but not those from COS-7 kidney cells. The mutation of this motif, which abolishes suppressing activity in the native context, also abolishes binding. The presence and activity of this suppressor in cells of the osteoblast lineage suggest that it is expressed with other cell-specific transcriptional regulators of the osteocalcin gene, coordinately regulating expression of this gene in bone cells. PMID- 8718895 TI - The recent history of the autopsy. PMID- 8718897 TI - College of American Pathologists Conference XXIX on restructuring autopsy practice for health care reform. Let's make this autopsy conference matter. PMID- 8718896 TI - High autopsy rates at a university medical center. What has gone right? AB - OBJECTIVE: Identification of factors that contribute to a high autopsy rate at our institution. DESIGN: An objective analysis of the Autopsy Service's organization, functions, and process flow to identify factors that impact the autopsy rate. SETTING: The University of Texas Medical Branch at Galveston, Tex. METHODS: Statistics were collected using the autopsy log book and computerized data bank. The events starting at the time of a patient's death through the completion of the autopsy report were reviewed. The role of each significant event was analyzed. RESULTS AND CONCLUSIONS: The annual autopsy rates between 1981 and 1995 ranged between 45% and 59%. The largest number of nonmedicolegal autopsies were received from the Department of Internal Medicine. The Texas Department of Criminal Justice Hospital contributed the largest number of medicolegal autopsies. We conclude that the following key factors contribute to our high autopsy rates: (1) the organization and multiple functions of the Autopsy Service, particularly the presence of a Decedent Affairs Office, dedicated resident assignments, and internal and external quality control of the autopsies; (2) close interactions with clinicians, including timely communication of autopsy results to clinicians and a fostering of positive attitude among clinical residents and faculty; and (3) other factors such as the contributions to hospital risk management, disproving the idea that there may be increased litigation related to high autopsy rates, and support by the hospital administration. PMID- 8718898 TI - The autopsy. Some ethical reflections on the obligations of pathologists, hospitals, families, and society. AB - Since the 18th century, the autopsy has been seen as an essential tool for advancing medical knowledge and monitoring the quality of patient care. In recent years, based on the mistaken assumption that newer diagnostic antemortem procedures, especially imaging techniques, have rendered the autopsy obsolete, it has fallen into serious disuse and disrepute. Data disproving this assumption are now emerging. The autopsy is still needed as a final test of the newer diagnostic techniques and as a means for integrating anatomic, physiologic, biochemical, and clinical features of new, as well as established, diseases. This essay examines the sources of clinicians' and pathologists' moral obligations to obtain, perform, and attend autopsies. It underscores the ethical responsibilities of families, hospitals, and society to restore the autopsy to a place of importance in the elaboration of the natural history of human disease. PMID- 8718899 TI - Maximizing the benefits of autopsy for clinicians and families. What needs to be done. AB - The benefits of autopsy, both for clinicians and families, are reviewed. The autopsy rate in the United States has fallen dramatically in the past 50 years. The many factors contributing to this decline are summarized. For clinicians and families to receive the maximum benefits from the autopsy will require (1) altering methods of obtaining consent (eg, using trained autopsy advocates and enhancing awareness of cultural issues); (2) altering autopsy procedures (eg, decreasing turnaround time; discussing issues and concerns with clinicians; and issuing more detailed, less technical reports); (3) improving communication with clinicians and families (eg, collecting clinician and family contact information on autopsy permits; inviting attendings, housestaff, and students to attend gross conferences; making telephone calls regarding unexpected findings; guaranteeing reports to clinicians; writing nontechnical summary letters to clinicians and families when the final report is completed; and conducting postautopsy conferences); and (4) educating both medical professionals and public citizens about the value of autopsy (eg, featuring autopsy results in medical conferences, distributing educational materials, and using print and electronic media). PMID- 8718900 TI - Epidemiologic and clinical challenges in reviving the necropsy. PMID- 8718901 TI - The role for regional autopsy centers in the evaluation of covered deaths. Survey of opinions of US and Canadian chairs of pathology and major health insurers in the United States. AB - OBJECTIVE: To evaluate the advantages and disadvantages of, as well as the attitudes of health care professionals and insurers toward, the development of regional autopsy services. DESIGN: Survey of 150 medical school departments of pathology in the United States and Canada and 12 representative major health insurers in the United States. RESULTS: Of the 25 respondents from the pathology departments, most were in favor of regionalization of autopsy services, if properly underwritten. Of the five respondents from the health insurers, most were disinterested in the autopsy as a measure of outcome and unwilling to provide support. CONCLUSIONS: Health care is being regionalized around networks of insurers rather than hospitals. The networks are defined by a mixture of hospitals, physician groups, and other health care professionals. Within networks, the goal is to subscribe groups of patients, covered lives, for all medical needs from primary to complex care. As the economic risk of caring for patients is shifted to physicians, the incentive to provide service at the lowest possible cost grows, as does the need to assure that medical mismanagement does not occur. To provide quality care at affordable costs, it is necessary that outcomes, including deaths, be professionally evaluated. The present system of death investigation involves hospital colleagues and is potentially biased. Regional autopsy centers that provide timely expert information should be part of the health care system. Medical schools are potential sites for regional autopsy programs because they have the personnel needed to conduct appropriate death related studies. Most schools are affiliated programmatically and economically with surrounding hospitals and physicians in a manner in which outcomes, costs, and quality of clinical service are of common interest. PMID- 8718902 TI - A potpourri of legal issues relating to the autopsy. PMID- 8718903 TI - Relating autopsy requirements to the contemporary accreditation process. AB - The Joint Commission on Accreditation of Healthcare Organizations has long been a strong proponent of the autopsy. Standards related to autopsies are clearly set forth in the current manual for hospitals. However, radical changes in the health care environment and in the philosophy of accreditation are placing the use of autopsy results in a new context. In that context, autopsy results are competing with other performance measures as drivers of quality improvement, and the autopsy itself is competing for scarce resources. A successful outcome will require autopsy advocates to persuade key audiences (viz, consumers, purchasers, and hospital executives) that autopsy findings lead to substantive improvements in the delivery of care, and that the autopsy is more worthy of support than other sources of performance data. PMID- 8718904 TI - Autopsy. A crucial component of human clinical investigation. AB - OBJECTIVE: To determine the proper role of human clinical investigation, including autopsy, in public health research policy. DATA SOURCES: Medical reports and reviews, and literature concerning the philosophy of medicine. CONCLUSIONS: Autopsy has always been a cornerstone of medical research. Although many pathologists appreciate the research value of autopsies, many other physicians do not. Declining federal support for human clinical research in general, and autopsy in particular, party reflects the popular view that laboratory experimentation-especially that performed on nonhuman animals-can reliably model human conditions. Human clinical research is often difficult, expensive, and time-consuming, but there seems to be no adequate substitute for studies of naturally occurring human conditions. Such studies address many critical medical questions. Autopsy remains an invaluable complement to evolving molecular biology techniques. PMID- 8718905 TI - The role of managed health care organizations in autopsy reimbursement. PMID- 8718906 TI - The need for a national autopsy policy. PMID- 8718908 TI - Expanded programme on immunization (EPI). Measles control in the WHO African region. PMID- 8718907 TI - A prototype Internet autopsy database. 1625 consecutive fetal and neonatal autopsy facesheets spanning 20 years. AB - OBJECTIVE: To demonstrate that cause-of-death statements can be generated by a computer algorithm from an autopsy database composed of diagnostic terms. DATA SOURCES: Over 49 000 autopsy facesheets contributed by over a dozen institutions were collected from a publicly accessible Internet autopsy database. This database is available at the following web site: http:@www.med.jhu.edu/pathology/iad.html STUDY SELECTION: To test the feasibility of creating and using a publicly available autopsy database, and to identify the technical and medicolegal problems that may arise with such a novel resource, a prototype study was designed by selecting autopsy facesheets from fetal and neonatal deaths. An algorithm was developed to determine the cause of death from the listing of anatomic diagnoses. DATA EXTRACTION: One thousand six hundred twenty-five fetal and neonatal autopsy facesheets were selected encompassing fetal and neonatal deaths occurring up to 28 days after birth. DATA SYNTHESIS: The algorithm determined causes of death from autopsy facesheet data in all cases. On review by an experienced pediatric pathologist, these automatically generated cause-of-death statements required no modification or only slight modification in over 90% of cases. CONCLUSIONS: A large multi-institutional autopsy database composed of demographic and diagnostic information has been deposited on the Internet. This information can be freely downloaded and used by any researcher without violating patient confidentiality. As a demonstration of one possible application of the database, fetal and neonatal autopsies generated cause-of-death statements using a computer algorithm. One can anticipate that the wealth of information contained in autopsy facesheets can be assembled into a database that will serve the public interest. PMID- 8718909 TI - Sympathetically-maintained pain related to reproductive surgery. PMID- 8718910 TI - Prevalence of symptoms among patients with advanced cancer: an international collaborative study. Symptom Prevalence Group. AB - The aims of this study were (a) to estimate the prevalence of pain and eight other common symptoms in a large population of patients with advanced cancer from different palliative care centers, and (b) to assess the differences in prevalence of the symptoms by primary site. In 1990-1991, the prevalence of eight major symptoms and performance status were assessed prospectively among 1840 cancer patients in seven hospices in Europe, the United States, and Australia. The data were collected at each institution using structured data collection sheets from the World Health Organization's (WHO) Cancer and Palliative Care Unit. The prevalence of moderate to severe pain was 51%, ranging from 43% in stomach cancer to 80% in gynecological cancers. Nausea was most prevalent in gynecological (42%) and stomach (36%) cancers, and dyspnea (46%) in lung cancer. There were statistically significant differences in the prevalence of most symptoms depending on the primary site of cancer and the hospice. Population based follow-up studies are needed to document the incidence and prevalence of symptoms throughout the course of the disease. PMID- 8718911 TI - Effects of induced metabolic alkalosis on perception of dyspnea during flow resistive loading. AB - Treatment of dyspnea in patients with advanced cancer is an important issue. The purpose of the present study was to assess the effect of induced-metabolic alkalosis produced by administration of sodium bicarbonate on dyspneic sensation. In seven healthy subjects, dyspnea was induced by having them breathe with a flow resistive load (24 cm H2O/L/sec) for 6 min before and after administration of sodium bicarbonate (0.5 mmol/kg, per os+2 mmol/kg, IV). The intensity of dyspnea was rated using a visual analogue scale (VAS). The VAS scores and minute ventilation during loaded breathing after administration of sodium bicarbonate were significantly lower than those before administration of sodium bicarbonate. These results indicate that induced metabolic alkalosis may alleviate the intensity of dyspneic sensation by a reduction in ventilatory drive. PMID- 8718912 TI - Use of red blood cell transfusions in terminally ill cancer patients admitted to a palliative care unit. AB - Anemia is often associated with neoplastic disorders. Blood transfusions are used to alleviate the discomfort of anemic cancer patients. Of 246 terminally ill cancer patients admitted to our palliative care unit from October 1991 to December 1993 (128 women and 118 men), 31 patients (12.6%) (17 men and 14 women; age, 69.5 +/- 12 years) received on average 2.8 units of packed red blood cells (PRBCs) (range, 2-7 units/patient) in 35 separate admissions. PRBCs were transfused in the presence of low hemoglobin (Hb) levels ( < or = 8 g/dL) and/or severe fatigue or dyspnea. Pre-transfusion performance status, cognitive function, dyspnea, and fatigue at rest (evaluated by a four-point scale), complete blood count, serum albumin, and C-reactive protein were determined. The day after transfusion, subjective well-being was recorded as "yes/no" improvement in comparison with the pre-transfusion day. Improved subjective well-being after blood transfusion was reported in 51.4%, without significant relationship to pre transfusion Hb levels or performance status. The influence of blood transfusion on subjective well-being was not related to the severity of dyspnea or fatigue. Twenty-one patients (60%), including seven with subjective improvement, died during the same hospitalization, a median of 49 days after transfusion. Pre transfusion Hb level did not differ significantly in patients who benefited and did not benefit from transfusion, whereas time before death was significantly (P < 0.001) shorter in patients who did not benefit. In the discharged patients (40%), the median interval between transfusion and discharge was 13 days and the frequency of subjective improvement in well-being was 78.6%. Our data suggest that two main areas should be investigated, namely the relation between low Hb levels and symptoms and signs in terminally ill cancer patients, and the correct timing for effective blood transfusion. A combination of criteria is needed for effective transfusion; they must include not only Hb levels but also type and severity of anemic symptoms and signs. Furthermore, the identification of reliable prognostic indicators for survival would be useful. PMID- 8718913 TI - The impact of a hospice consultation team on the care of veterans with advanced cancer. AB - To address the needs of patients with advanced cancer, all Veterans' Affairs hospitals were asked to establish teams of clinicians whose focus was to deliver expert palliative care. At the Philadelphia Veterans' Affairs Medical Center, our Hospice Consultation Team brought together professionals who had worked together informally in some of the outpatient oncology clinics. We conducted a prospective study of all consults received during the first year of the team's operation to determine whether or not the team approach would duplicate already available services or provide improved care. We identified 164 new medical/ nursing problems in 75 patients, 85% of which we were able to resolve. There were 152 new psychosocial/spiritual problems, but only 40%-61% were resolved. Of 22 patients followed in oncology clinics, new problems were identified and resolved in 21. We conclude that the expertise of the Hospice Consultation Team members along with the team process provided improved care to these patients. PMID- 8718914 TI - Sedation for symptom control in Japan: the importance of intermittent use and communication with family members. AB - We reviewed the circumstances surrounding the use of sedation for symptom control in a Japanese hospice. Of 143 inpatients, 69 (48.3%) received sedation and died an average 3.9 days after sedation was begun. Symptoms requiring sedation included dyspnea, pain, general malaise, agitation, and nausea. In 83% of cases, those symptoms were escalating as death approached. In about one-half of the cases, sedation was carried out intermittently until the patient died. Sedation was gained by such sedatives as midazolam, morphine, and haloperidol. Side effects included suppression of the respiratory and/or circulatory system in nine cases (in four cases it caused death), and delirium in one case; tolerance and dependence were also observed in two cases. We also examined the explanation to and understanding of the patients and their family members about sedation. This experience suggested the type of communication methods that are likely to be useful in Japan. It stresses the importance of intermittent use of sedation and communication with family members. We propose criteria for sedation to improve symptom control that would be acceptable in Japan. PMID- 8718915 TI - Italian consensus on a curriculum for volunteer training in oncology. AB - Volunteers' education, together with selection and organization, are fundamental for an adequate volunteer service in oncology, which is focused on patient and family needs and is fully integrated with the activity of the care team. To achieve such an aim, well-defined requirements and educational objectives are essential. A curriculum for voluntary service in oncology was developed through a consensus of experts. By comparing the work carried out by six different teams, a questionnaire was drafted, which grouped 91 educational aims. For each, the questionnaire queried whether the aim was relevant or not, and, if yes, whether it should be pursued through (a) knowledge, (b) skill, and/or (c) attitude (with a possible multiple answer). To improve comprehension, the questionnaire was divided into five sections: (a) the association; (b) the disease, therapies, and care site; (c) voluntary service in oncology; (d) psychosocial aspects of the patient and family; and (e) approach, communication, and acceptance. The questionnaire was sent to 117 delegates from 93 Italian volunteer organizations who, during a period of 8 years, had attended a course for volunteer trainers held by the Italian League Against Cancer. As indicated in the letter forwarded with the questionnaire, the final document would report only the aims considered relevant by at least 70% of the participants. Moreover, the area(s) of pursuit (knowledge, skill, and attitude) would be stated for each goal when indicated by at least one-half of the participants. Of 117 individuals questioned, 83 (71%) replied. Of the 91 aims proposed, 60 were considered relevant, and a type of approach (knowledge, skill, and/or attitude) was suggested. The formative curriculum of voluntary service in oncology represents a consensual guideline for future training programs. PMID- 8718916 TI - Intravenous hydration for terminal patients: what are the attitudes of Israeli terminal patients, their families, and their health professionals? AB - Dehydration in terminally ill patients is a common condition and its treatment in an active oncology unit often involves intravenous (IV) hydration programs. The decision to use IV hydration is only partly determined by an objective assessment of the clinical data. The other component is the attitudes of those involved in the decision-making process. This pilot study surveyed the attitudes of patients, family members/friends, nurses and doctors toward IV hydration in this patient population. Although the majority of the conscious patients (95%), family members/friends (81%), and nurses (64%) were not involved in the decision-making process, the expressed attitudes were generally positive. The three most commonly stated reasons for IV therapy were "medication," "giving fluids" and "giving morphine." Interestingly, among the reasons expressed, the amelioration of thirst was never explicitly cited. PMID- 8718917 TI - Using pressure to decrease the pain of intramuscular injections. AB - The purpose of this study was to determine if applying pressure to the site for 10 sec prior to an intramuscular injection would reduce injection pain, an approach suggested by anecdotal observation and the gate control theory. The subjects were 93 patients who had dorsogluteal intramuscular injections of immune globulin at a county health department. Forty-eight received the pressure treatment and 45 received a standard injection in which no pressure was applied. Mean pain intensity on a 100-mm visual analogue scale, adjusted for differences in injection volume, was 13.6 mm for the experimental group and 21.5 mm for the control group (P = 0.03). The findings suggest that simple manual pressure applied to the site is a useful technique to decrease injection pain. PMID- 8718918 TI - Midazolam for intractable hiccup. AB - We describe two terminally ill cancer patients severely distressed by intractable hiccup. Commonly recommended treatments were ineffective. Intravenous midazolam 5 10 mg rapidly relieved the hiccup. Maintenance treatment comprised midazolam 40 120 mg/24 hr by continuous subcutaneous infusion. PMID- 8718919 TI - [Research and post-graduate studies]. PMID- 8718920 TI - [Antineutrophil cytoplasmic antibodies (ANCA): study of their presence in diseases not associated with arteritis]. AB - Our study describes the presence of antineutrophil cytoplasmic antibodies (ANCA) in a group of different pathologies comprising 101 patients. Rheumatoid arthritis, systemic lupus erithematosus, idiopatic neutropenia, acute post streptoccocal glomerulonephritis, minimal change nephrotic syndrome, Downs syndrome, adult periodontitis, tumoral calcinosis, monoartheritis and lipodystrophy were investigated for ANCA, through indirect immunofluorescence and an indirect solid-phase immunoassay (ELISA). Our results show the pattern of distribution of ANCA in the diseases investigated, and allowed us to make the first description of ANCA in diseases such as Downs syndrome, acute post streptococcal glomerulonephritis and adult periodontitis. The high percentage of reactivity for ANCA detected in adult periodontitis, raise important questions about the possibility of reporting inaccurate percentages of positivity for some diseases, due to the presence of a concurrent disease such as adult periodontitis. PMID- 8718921 TI - [Immunologic characteristics of undernutrition. I. The undernourished patient in nutritional recovery]. AB - Malnutrition in children is a well known critical factor that determines immunocompetence changes with altered immune response and higher risk to many diseases, especially in developing countries. Moreover, it is related to increased morbi-mortality rates mainly due to infections. For those reasons, 12 undernourished children, age 5 to 24 months were studied along 8 weeks at the Nutritional Recovery Center of Chiquinquira Hospital in Maracaibo, Venezuela. There were 5 cases of kwashiorkor, 5 marasmatics, 1 mixed marasmus/kwashiorkor and 1 case with moderate malnutrition. After a control blood sample was taken and cutaneous tests were done, a nutritional recovery program was began. At regular time intervals and at the end of the study, tests were done again by measuring seric immunoglobulins (IgG, IgA, IgM), secretory IgA (IgAs), C3 and C4 complement, lymphocytic sub-populations, and auto antibodies; cutaneous hipersensitivity tests were also done. As a control group, 10 apparently healthy children of matching age and sex were also studied with the same parameters. Results show that basal seric Igs did not differ significantly from the control group and did not change along the recovery program period, but there was a significant decrease in IgAs at all times of the study. C4 did not change and C3 was lower than control (p < 0.05) but returned to normal value at the end of the recovery period. CD3 and CD4 lymphocytes showed the same pattern. Only two patients showed positive skin tests and auto antibodies were not detected. It is concluded that there is indeed an altered immune competence with low levels of C3, IgAs, and CD3-CD4 lymphocytes that is reversible after nutritional recovery. PMID- 8718922 TI - Plasma excitatory amino acids in autism. AB - Plasma amino acid levels were measured by high pressure liquid chromatography (HPLC) in fourteen autistic children, all below 10 years of age. Mean glutamic and aspartic acid valued were elevated (169 +/- 142 uM and 22.1 +/- 13 uM respectively) together with taurine (90.1 +/- 78.7 uM) (p > 0.1). All affected children had low levels of glutamine (241 +/- 166 uM; p < 0.01) and asparagine (22.9 +/- 12.9 uM; p < 0.01) as compared to normal values (585 +/- 25 and 59.2 +/ 4.2 uM respectively); eleven children had increased aspartic acid and eight children had high levels of glutamate; seven of these children had a concomitant increment of taurine. The increment of the three above mentioned compounds was observed at the same time only in five children. These findings demonstrate that abnormal plasmatic levels of neurotransmitter amino acids may be found in some autistic children. Increased glutamatemia may be dietary in origin or may arise endogenously for several reasons, among others, metabolic derrangements in glutamate metabolism perhaps involving vitamin B6, defects or blockage of the glutamate receptor at the neuronal compartment, or alterations in the function of the neurotransmitters transporters. Increments of taurine, an inhibitor, is likely compensatory and calcium dependent. PMID- 8718923 TI - Ketamine reduces lethality on the acute ammonia intoxication in mice. AB - Injection of large doses of ammonia (1.2g/kg, i.p.) was used to induce acute toxicity in mice which was characterized by hyperresponsiveness, taquipnea, clonic and tonic seizures and death. Pretreatment with 20, 40, or 80 mg/Kg, i.p., of ketamine increased 30 to 55% survival rate. This pretreatment significantly retarded the beginning of the first tonic convulsion attenuating its intensity and delayed the time of the animal death; but did not alter the onset of the first clonic seizures. These experiments may be an evidence that support the hypothesis that seizures due to hyperammonemia involve activation of excitatory amino acid receptors. PMID- 8718924 TI - [Asymmetric crying facies syndrome: report of 16 clinical cases]. AB - The Asymmetric Crying Facies (ACF) is included among the congenital facial palsy in childhood. We report the clinical characteristics, congenital anomalies and other associated disorders in sixteen patients with ACF examined from 1984 to 1995 in the pediatric units of the Hospital Adolfo Pons, Universitario and Clinico of Maracaibo, Venezuela. Statistically the side of lesion and the sex were independent. In seven of sixteen patients (43.75%) we found congenital anomalies and/or neurological disorders, being diaphragmatic hernia and the hyperkinesia a rare association no reported up to now in the reviewed literature. We suggest autosomal dominant inheritance with reduced penetrance in three of sixteen families. The other patients were sporadic cases. PMID- 8718925 TI - Changes in D-aspartic acid in human deciduous teeth with age from 1-20 years. AB - The racemization ration (D/L ratio) of aspartic acid in the dentin (collagen) of deciduous teeth was studied, using dentin of permanent teeth as a reference. D aspartic acid in deciduous teeth was found to increase almost linearly with aging. The reaction velocity constant of racemization (kyr-1) in deciduous teeth (1.1915 x 10(-3)yr-1) was approximately double the value for permanent teeth (6.2550 x 10(-4)yr-1). The velocity of the racemization reaction was similar between deciduous teeth and permanent teeth in a dry state. The D/L ratio determined using transverse sections of deciduous teeth tended to be high in the crown and low in the root. The reaction velocity of racemization is considered to be affected to some extent by the tooth material, ambient temperature and salivary pH, but most importantly by physiologic resorption of deciduous teeth at the root associated with age. PMID- 8718926 TI - Effect of trimethylamine-carbomethoxyborane on growth traits and lipid metabolism in lines of mice selected for high and low fat content. AB - The purpose of this study was to determine the effects of the amine carboxyborane, trimethylamine-carbomethoxyborane (B), on growth, body composition and lipid metabolism in lines of mice differing in fat content as a result of directional selection: high fat content (HF); low fat content (LF); and random control (RC). Mice were injected i.p. daily with either 20 mg/kg of B dissolved in 1% (w/v) carbomethoxycellulose as the vehicle or just vehicle (C) from 26 to 42 days of age. Growth rate, feed intake, feed efficiency, epididymal fat pad weight/BW and percentage body water were not affected by B treatment, but liver weight/BW was increased (P < 0.001). HF mice had larger epididymal fat pad weight/BW and less percentage body water when compared to LF mice (P < 0.001). Treatment with B lowered (P < 0.01) serum triglyceride and cholesterol levels, and selection for divergence in fat content led to positive divergence (P < 0.01, (HF-LF) > 0) in serum triglyceride and serum cholesterol levels; divergence x treatment interactions (P < 0.01) were due to administration of B eliminating the line differences present when C was administered. Treatment with B elevated (P < 0.001) HDL cholesterol level and lowered (P < 0.001) chylomicron, LDL and VLDL cholesterol levels, while positive divergence was found for all four fractions. Fecal triglyceride and cholesterol levels were increased (P < 0.01) by administering B, but were not affected by selection. Hepatic enzyme activity of cholesterol-7 alpha-hydroxylase was elevated (P < 0.001) by treatment with B while activities of other hepatic enzymes were depressed, e.g., acyl-CoA cholesterol acyltransferase. Line differences in activity of several hepatic enzymes also were found. These results indicate that B acts as a hypolipidemic agent in both high-fat and low-fat mouse genotypes, lowering serum cholesterol and triglyceride levels. However, at the administered dose, B had no affect on growth rate, feed intake, feed efficiency or body fat content. PMID- 8718927 TI - Associations between atlantoaxial and craniomandibular anatomy. AB - Roentgen-cephalometric studies have shown that anatomical features of the craniocervical junction are associated with head posture, cranial base angulation, and mandibular shape and growth. The aim of the present work was to study relationships between atlantoaxial and craniofacial morphology based on macroscopical observations of skeletal material derived from 38 prehistoric Polynesian and 53 prehistoric Thai people. The two uppermost cervical vertebrae and the mandible of each individual were studied macroscopically and each skull base was analyzed on cephalometric X-rays in lateral projection. The height of the atlantal anterior and posterior arches displayed a significant negative correlation with the cranial base angulation, in that the higher the arches the steeper the flexure between the sphenoidal/clival and clival/foraminal planes. None of the axial variables were associated with the cranial base angulation. The height of the atlantal posterior arch was also associated with the mandibular length, ramal height and gonial angle. Thus, in general, a high arch was seen in conjunction with a long, high and square shaped mandible, whereas a low arch was usually found together with a short and low mandible characterized by an obtuse jaw angle. The anterior height of the axis (vertebral mass + dens) was significantly associated with mandibular length and ramal height. It is suggested that these results are evidence of the intimate ontogenetic development of the atlas and the cranial base, and a reflection of the functional relationship between atlas and cranium. PMID- 8718928 TI - Skeletal muscle growth of oMTla-oGH transgenic mice. AB - Forty-eight transgenic mice carrying an ovine metallothionein 1a-ovine growth hormone (oMTla-oGH) transgene and 48 littermate control mice were used to investigate the effect of GH transgene on the growth and biochemical characteristics of skeletal muscle. Transgene expression was initiated in the transgenic mice by the addition of zinc sulfate to the water at 21 d of age; control mice were also supplemented with zinc sulfate. These mice were maintained on zinc sulfate until 84 d of age. Groups of mice (16 controls, 16 transgenics) were killed at 21, 42 and 84 d of age and muscles from the hind leg were dissected, weighed and analyzed. At 84 d, male transgenics were 32% heavier than controls, while female transgenics were 47% heavier. Transgenic mice of both sexes had smaller (p < 0.01) muscles than controls at weaning (21 d). In spite of significantly heavier body weights of transgenic mice at 84 d of age, there were no significant differences in muscle weights. This was due to a significantly lower (p < 0.01) proportion of muscle, expressed as percentages of body weights, in transgenic mice compared with controls. Higher DNA and RNA concentrations at 42 d of age and elevated cathepsins C and H activities at 42 and 84 d of age indicate that muscle protein metabolism is more active in transgenic mice, which are growing at a greater rate than controls from weaning to 84 d of age. The fact that oMTla-oGH transgenic mice inherently have a lower proportion of muscle, compared to controls and that this proportion does not change in spite of transgene activation and 30 fold increase in plasma GH levels, suggests the hypothesis that muscle growth may be controlled by locally produced IGFs, which are essentially independent of circulating GH concentrations. PMID- 8718929 TI - Monocyte activation on titanium-sputtered polystyrene surfaces in vitro: the effect of culture conditions on interleukin-1 release. AB - The release of interleukin-1 alpha (IL-1 alpha) by human peripheral blood monocytes cultured for 24 and 48 h on polystyrene (PS) and titanium-sputtered polystyrene (Ti) was evaluated. Magnetron sputtering of the PS surfaces resulted in a formation of a 50-nm-thick coat, consisting of an outer layer of TiO2. Monocytes released IL-1 alpha without the addition of exogenous stimuli. A doubling of the culture time from 24 to 48 h did not have a major effect on the amount of IL-1 alpha released. The IL-1 alpha levels were increased by addition of lipopolysaccharide (LPS). High concentrations of PS particles (1 and 3 microns diameter) were equally effective stimuli for IL-1 alpha release as LPS. Preadsorption of fibronectin to culture plates augmented LPS-stimulated IL-1 alpha secretion, whereas preadsorbed fibrinogen had an inhibitory effect. Our observation indicate a direct activation of monocytes by PS and Ti, resulting in IL-1 alpha secretion, which is modified by protein adsorption and exogenous stimuli. PMID- 8718930 TI - Adsorption of bovine serum albumin on to titanium powder. AB - The adsorption of bovine serum albumin (BSA) on to titanium powder has been studied as a function of protein concentration and pH, and in the presence of calcium and phosphate ions. Isotherm data have shown that the adsorption process does not follow the Langmuir model (inflection points). The maximum adsorption (Admax) at pH 6.8 is 1.13 +/- 0.21 mg m-2. For the pH dependence of adsorption the amount adsorbed increases with decreasing pH (at pH 5.15 Admax = 1.31 +/- 0.2 mg m-2), indicating that hydration effects are important. Adsorption increases and decreases in the presence of calcium (at pH 6.8 for 0.002 M Admax = 1.73 +/- 0.23 mg m-2) and phosphate (at pH 6.8 for 0.002 M Admax = 0.64 +/- 0.14 mgm 2)ions, respectively, indicating that electrostatic effects are important. The time dependence, isotherm and desorption data provide indirect evidence of possible conformational changes in the BSA molecule. PMID- 8718931 TI - Comparison of the size and morphology of UHMWPE wear debris produced by a hip joint simulator under serum and water lubricated conditions. AB - This paper investigates the effect of lubricant on the size and morphology of ultra-high molecular weight polyethylene (UHMWPE) wear debris produced by a hip joint simulator. Bovine serum and water were chosen as the lubricants for comparison. When bovine serum was used as lubricant, the majority of the wear debris produced was submicron sized and equiaxed or elongated in shape; no transfer film was found on the femoral heads. When water was used as lubricant, large pieces of thin UHMWPE flakes about 2-3 mm across were produced and patches of transfer films were observed on the femoral heads. These large flakes were comprised of numerous smaller thin flakes compacted together. While the size and shape of the wear debris produced with serum lubrication are similar to those reported from clinical studies in total hip replacement, the debris produced with water lubrication bears no resemblance to what has been seen clinically. PMID- 8718932 TI - Unacceptable biodegradation of polyethylene in vivo. AB - The chemical characterization of 19 retrieved ultra-high-molecular-weight polyethylene tibial plateaux, six new ones and one raw bar was performed by means of infrared spectroscopy. The surface and bulk oxidation and biodegradation indexes were calculated. The raw bar has a measurable oxidation, which increases on the bulk and on the surfaces of the new plateaux. In the retrieved plateaux, the average oxidation index increases further both on the bulk and on the surfaces; the worse values were present on the worn area. Similar results were found for the biodegradation index. The data show that the biotic in vivo degradation is promoted by the oxidation present on the new plateaux and that it occurs through a different mechanism, abiotic thermal-, photo-, gamma-radiation oxidation, evaluated by the oxidation index. PMID- 8718933 TI - Human chondrocytes proliferate and produce matrix components in microcarrier suspension culture. AB - Chondrocytes propagated in monolayer culture proliferate and change into 'fibroblastoid'-like cells. This change is characterized by a shift in production of collagen type II to I and from high- to low-molecular-weight proteoglycans. When propagated in three-dimensional culture, chondrocytes have limited ability to divide but re-express their original characteristics. The goal of the present study was to determine whether a microcarrier suspension culture system would support chondrocyte proliferation and phenotype expression. Our experiments indicate that a collagen type I microcarrier (cellagen) best supported chondrocyte proliferation and phenotype expression. Cells in cellagen microcarriers multiplied at least twentyfold within 2 weeks and had doubling times of 2 to 3 d. Viable and metabolically active cells were retrieved with ease. The harvested chondrocytes had no detectable staining for collagen type I and stained intensely for collagen type II. Our studies demonstrate that the microcarrier suspension culture system supports growth and enhances expression of the 'chondrocytic' phenotype. Attachment to a constrained surface and the fluid shear forces on the microcarriers during suspension culture may have helped chondrocytes to reacquire their rounded shape and produce cartilage matrix components. PMID- 8718934 TI - Modulation of the dissolution profiles from Geomatrix multi-layer matrix tablets containing drugs of different solubility. AB - A new multi-layer tablet design has recently been proposed for constant drug release: Geomatrix Technology (Jago Pharma, Muttenz, Switzerland). It consists in the application of a drug-free barrier layer on one or both bases of an active core (hydrophilic matrix). The partial coating modulates the core hydration process and reduces the surface area available for drug release. The result is an extended release that draws close to a linear profile. The device was mainly intended for soluble drugs, while an excessive reduction of the release rate may be obtained with drugs of low solubility. In this study a new time-dependent polymeric barrier is proposed to control the release of sparingly soluble drugs. Two different barrier compositions (one swellable and one erodible) are applied on active cores containing drugs of different water solubility, Trapidil, Ketoprofen and Nicardipine hydrochlorides, and the drug dissolution patterns of the different multi-layer devices are compared. During dissolution, the swellable barrier swells and gels, but is not eroded, thus acting as a modulating membrane during the release process. The erodible barrier, instead, is progressively removed by the dissolution medium, exposing in time an increasing extent of the planar surface(s) of the core to interaction with the outer environment and to drug release. Both types of coatings are able to control drug release from the devices: the swellable barrier shows a stronger modulation efficiency and is more suitable to modify the delivery pattern of highly soluble drugs; the erodible barrier shows a time-dependent coating effect that provides better control of the dissolution profile of sparingly soluble drugs. PMID- 8718935 TI - Purity and stability assessment of a semi-solid poly(ortho ester) used in drug delivery systems. AB - The research work carried out for developing bioerodible drug delivery devices in which the erosion process was to be confined to the polymer-water interface is at the origin of the discovery of a class of polymers known as poly(ortho esters) (POEs). Thus far, three POE systems have been described. The latest POE was prepared by a transesterification reaction between a triol and an ortho ester, followed by a self-condensation of the reaction product. This polymer, which exhibits viscous characteristics at room temperature, was investigated for use as a drug delivery system in glaucoma filtering surgery. The assessment of POE purity and stability was carried out by a detailed analysis of the influence of the purification procedure and storage conditions. This bioerodible semi-solid POE was purified by a repeated precipitation procedure. Elimination of the small molecular weight oligomers and monomers and of the catalysts and stabilizers used in the synthesis, as well as a decrease of the polydispersity, were obtained with this method. Fourier transform infrared analysis also verified the disappearance of degradation products after the first precipitation. Drying of the precipitated polymer was performed at 40 degrees C in order to avoid thermal degradation of the POE at higher temperatures and to facilitate solvent evaporation through the polymer network by a reduction of polymer viscosity. Water vapour uptake of the polymers stored at different relative humidities has demonstrated the high moisture sensitivity of these semi-solid POEs. The average molecular weight of the polymer and hence its viscosity, as well as the solubility characteristics of the incorporated drug, were found to have a considerable influence on the rate of water vapour absorption and on polymer degradation. The use of inert gas or vacuum to maintain the polymer under anhydrous conditions has been studied. Storage of the semi-solid POE under argon in sealed glass bottles provides good protection of the polymer over time. PMID- 8718936 TI - Heparin-like functionalized polymer surfaces: discrimination between catalytic and adsorption processes during the course of thrombin inhibition. AB - Thrombus formation on blood-contacting artificial surfaces is a major problem. Antithrombogenic polymer surfaces have been obtained either by heparin binding, or by grafting sulphonate and/or amino acid sulphonamide groups on insoluble polystyrene. In addition to their capacity to adsorb thrombin, such surfaces were shown to be able to catalyse its inhibition by antithrombin III (AT), i.e. they are endowed with heparin-like activity. The results were mainly obtained by using clotting assays. In many cases, delineating adsorption and catalytic processes by such assays is not possible when evaluating anticoagulant polymer surfaces. To overcome this problem, the kinetics of thrombin adsorption and inhibitions by AT and heparin cofactor II (HC) in the presence of such surfaces have been measured by using an assay performed with a thrombin-specific chromogenic substrate. A simple kinetic model of thrombin consumption is proposed. The relevant calculations, carried out with the help of a computer program, lead to determination of relative second order rate constants of thrombin adsorption and inhibitions by AT and HC in the presence of the polymers. In addition to thrombin adsorption, polystyrene surfaces bearing only sulphonate groups catalyse inhibition by AT, whereas polystyrene surfaces bearing either aspartate, glycinate or isophthalate sulphonamide groups catalyse both inhibitions by AT and HC. PMID- 8718937 TI - Glass-ionomer cements based on poly(acrylic acid-co-vinyl alcohol) in drug release model formulations. AB - The release of methylene blue from glass-ionomer cements based on a commercial glass powder and partly hydrolysed poly(acrylic acid-co-vinyl alcohol) has been studied. A series of random copolymers were synthesized from acrylic acid and vinyl acetate monomers, using feed ratios of 10-90 mol% acrylic acid. The acetate group was hydrolysed using sodium hydroxide. According to proton nuclear magnetic resonance analysis, the degree of hydrolysis was about 40%. The residual Na+ content in the samples after hydrolysis was in the vicinity of 3 mmol g-1 polymeric sample. The viscosity average molecular weight of the copolymers was the region of 60,000 g mol-1 for samples with low acrylic acid content and in the region of 300,000 g mol-1 for samples with a larger fraction of acrylic acid. Cement sample discs were prepared and the porosity of the cement was determined using N2 adsorption-desorption isotherms. The specific surface area was 10 m2 g 1, and the volume of the pores (< 20 nm) 0.035 cm3 g-1. The swelling indices and the release rates in two different buffers, pH 2.0 and pH 7.4, were determined for fresh and dried samples. An acidic environment did not bring about significant swelling in the samples and release of the model agent at pH 2.0 was limited. At a physiological pH value the cements swelled in a hydrogel manner and the fractional release of model agent increased. PMID- 8718938 TI - Antioxidant N-acetyl-cysteine increasing cell adhesion capability could facilitate the biocompatibility processes. AB - Cell adhesion plays an important role in several cell processes and functions, including differentiation, proliferation and death. An important role for cell attachment to medical devices in biocompatibility studies has also been hypothesized. In this paper we report that the use of the antioxidant drug N acetyl-cysteine is capable of increasing the adhesion properties of epithelial cells in culture. This is associated with a modification of specific cytoskeletal element assembly, such as microfilament system molecules. In contrast, no quantitative alterations in the expression of certain surface receptors for extracellular matrix molecules, such as VLA2, VLA3 and VLA6, are found. These data seem to indicate that intracellular oxidative balance, in particular of thiol groups, could play a key role in the cell adhesion properties and that N acetyl-cysteine treatment, acting as 'thiol supply', could be of importance in several circumstances, including biocompatibility of medical devices. PMID- 8718939 TI - Cytocompatibility of Ti-6Al-4V and Ti-5Al-2.5Fe alloys according to three surface treatments, using human fibroblasts and osteoblasts. AB - Titanium alloys are well known for their superior mechanical properties as well as for their good biocompatibility, making them desirable as surgical implant materials. However, these alloys have been proven to behave poorly in friction since wear particles were often detected in tissues and organs associated with titanium implants. In this paper, three surface treatments were investigated in order to improve the wear resistance and the hardness of Ti-6Al-4V and Ti-5Al 2.5Fe: (a) glow discharge nitrogen implantation (10(17) atoms cm-2), (b) plasma nitriding by plasma diffusion treatment (PDT) and (c) deposition of TiN layer by plasma-assisted chemical vapour deposition (PACVD) additionally to PDT. Surface characterization after the different treatments showed considerable improvement in surface hardness, especially after the two nitriding processes. Moreover, the good corrosion resistance of untreated alloys was maintained. A cell culture model using human cells was chosen to study the effect of such treatments on the cytocompatibility of these materials. The results showed that Ti-5Al-2.5Fe alloy was as cytocompatible as the Ti-6Al-4V alloy and the same surface treatment led to identical biological consequences on both alloys. Nitrogen implantation did not modify at all the cellular behaviour observed on untreated samples. After the two nitriding treatments, cell proliferation and viability appeared to be significantly reduced and the scanning electron microscopy study revealed somewhat irregular surface states. However, osteoblast phenotype expression and protein synthesis capacity were not affected. PDT and PACVD may be interesting alternatives to the physical vapour deposition technique. PMID- 8718941 TI - Implications of the emerging home systems technologies for rehabilitation. AB - This paper gives a review of the current and anticipated developments in Integrated Home Systems and raises the implications these have in the field of rehabilitation. Firstly, the systems under consideration are defined and their development world-wide outlined. An overview of the European Home Systems Specification is given. The implications of this technology for rehabilitation are then raised considering: environmental control in the home; how people with degenerative conditions may be more readily able to remain in their homes as condition progresses; and remote health monitoring. The expectation that the initial impact will be in the area of environment control is stated, and a closer look at this field is made comparing current with anticipated systems. An outline is given of the TIDE Project HS-ADEPT (Home Systems--Access for Disabled and Elderly People to this Technology). The key findings of the survey of users made at the outset of the project, relevant to this paper, are given. The ongoing work of the project is described in brief. The continuing developments in Integrated Home Systems and in particular the issue of standards are noted. Concluding comments are offered in the hope of stimulating the necessary interdisciplinary discussion if this technology is to realize its full potential in the field of rehabilitation. PMID- 8718940 TI - 5-FU loaded pHEMA drainage implants for glaucoma-filtering surgery: device design and in vitro release kinetics. AB - Implantable monolithic and reservoir-like water-swellable drainage devices were developed for the subconjunctival sustained release of 5-fluorouracil (5-FU) in glaucoma-filtering surgery. A water-swellable matrix was formed of a copolymer of 2-hydroxyethyl methacrylate (HEMA) with different amounts of ethylene glycol dimethacrylate (EGDMA). Drug incorporation was done before polymerization and cross-linking. Briefly, to prepare the monolithic device the monomer-drug mixture is compression moulded into a 10 mm cylinder of 1 mm length. Furthermore, reservoir-like devices were obtained by coating the monolithic devices with a highly cross-linked polymer of HEMA (pHEMA) composition. The pHEMA devices containing 5-FU or not were well characterized by means of dynamic swelling studies, structural and thermal analysis. The release of 5-FU from these implants was studied in vitro. The rate of drug release was controlled by changing the drug loading (i.e. 10 mg or 20 mg 5-FU per device), cross-linking density of polymer matrix and type of implantable device, i.e. monolithic or reservoir-like device. While monolithic devices are releasing total releasable 5-FU during the first 10 h, reservoir-like devices prolong 5-FU release for up to 120 h. The 5-FU diffusion coefficient in swollen devices (Ds,s) is in the order of 10(-8) cm2 s-1 (approximately 10 times smaller than Dw,g values) and it is dependent on the cross-linking density of polymeric matrix and device load. These preliminary results suggested that 20 mg 5-FU-loaded reservoir-like devices may be a potentially effective system to deliver 5-FU into the subconjunctiva. PMID- 8718942 TI - From SAW to Comspec. AB - This paper details the development of two systems Switch Access to Windows (SAW) and Communicator Specification (Comspec). Descriptions are given of the two systems and their development methodologies. Convergence of approach both in the designer and user interfaces as well as the development methodologies arose naturally in the course of development despite starting with what seemed to be very different aims. SAW addresses a particular form of communication, that between a switch user and computer applications. Comspec will, as a comprehensive and expandable communication system, subsume SAW in the long run. PMID- 8718943 TI - Providing rehabilitation integrated systems using existing rehabilitation technology. AB - Achieving appropriate integration of control of multiple assistance devices for an individual is currently limited by available integrated systems. An overview of the provision process for Rehabilitation Integrated Systems (RIS) is presented which stresses the need for detailed assessment of the individual's abilities and requirements. A software system is described which integrates existing assistive applications on a PC in an attempt to provide a flexible RIS which can be tailored more precisely to an individual. This has required the use of both DOS and Windows applications and highlighted problems due to limitations of existing RISs, despite user requirements being taken fully into account. While Windows offers the opportunity to remove some compatibility problems, it is concluded that interfacing standards for both software and hardware are necessary to overcome the problems this approach engenders. For individuals who have to use 'intrinsically slow' user interfaces, it is identified that (i) the availability of quantitative comparisons of efficiency of selection methods and (ii) increased choice of selection methods is important. PMID- 8718944 TI - Integrating assistive technologies: current practices and future possibilities. AB - Many people with multiple impairments need to use a powered wheelchair, voice output communication aid, environmental control, and access information technology. Integrating and mounting such assistive devices on a powered wheelchair can be time-consuming, expensive and result in compromised performance. This paper offers a concept of assistive device integration, based around a continuum ranging from wholly discrete systems to fully integrated systems. Using practical examples, advantages and disadvantages of different approaches to integration along this continuum are identified, particularly with regard to control characteristics of different types of assistive technology. Limitations of current practices and technologies are discussed and measures suggested to enable systems to achieve better integration. PMID- 8718945 TI - An integrated approach to Smart House technology for people with disabilities. AB - It is now commonly accepted that 'Smart House' technology can play a significant part in helping both elderly and disabled people enjoy a greater degree of independence in the near future. In order to realize this aspiration, it is necessary to examine a number of factors: the development of the appropriate Home Bus technologies and supported devices; the development of the appropriate user interfaces that will allow people with a range of special needs use the system; the incorporation of the requirements of the 'Smart House' controller with the other technological needs of the user; and the development of mainstream technologies that will affect the cost and availability of devices to the user. This paper will examine the above points and suggest appropriate actions and trends. It will draw upon the work of a four-member consortium currently finalizing a technical development project under the EC TIDE program, the experience of research and commercial organizations engaged in development work in associated areas and the experiences of the Dublin-based, Central Remedial Clinic and in particular, its Client Technical Services Unit. The CTSU have been actively engaged in the development of systems for clients and direct clinical assessments for the last 12 years. PMID- 8718946 TI - Frictional action at lower limb/prosthetic socket interface. AB - The frictional action at stump/socket interface is discussed by a simplified model and finite element model analyses and clinical pressure measurements. The friction applied to the stump skin produces stresses within tissue and these stresses may damage the tissues and affect their normal functions. The combination of normal and shear stresses is considered to be a critical factor leading to amputee's discomfort and tissue damage. However, friction at the stump/socket interface has a beneficial action. A simplified residual limb model and a finite element model using real geometry have been developed to analyse the support action of friction. Both results show that the friction plays a critical role both in supporting the load of the amputee's body during the support phase of the gait cycle and in preventing the prosthesis from slipping off the limb during swing phase. Pressure at the below-knee socket during walking were measured with conditions of different friction. The results reveal that a larger pressures was produced at the lubricated interface than at the normal interface. A proper choice of coefficient of friction will balance the requirements of relief of load stress and reduction of slip with the general ability to support loads. PMID- 8718947 TI - Clarification of the mechanical behaviour of spinal motion segments through a three-dimensional poroelastic mixed finite element model. AB - The purpose of this study is to clarify the mechanical behaviour of spinal motion segments through a proper numerical model. The model constructed can give correct information and provide medical fields with valuable guidance in solving clinical problems occurring in the spine. A three-dimensional poroelastic finite element model of spinal motion segments is constructed and a mixed formulation is introduced. The geometry of the model is automatically formed from a series of CT scanning images. Vertebral column, intervertebral joint, facet joints and ligaments are all included in the model. The contact surface of facet joints is considered as the inclined boundary. Such inclination is imposed when the contact surface is under compression. Ligaments surrounding the vertebral body and the intervertebral disc are put into the model when they are under tension. Iteration is implemented in the computing process to meet such boundary characteristics of facet joints and ligaments. Prediction of the mechanical behaviour in the segment under long term creep loading, is demonstrated using the current algorithm. Results show that the model and corresponding numerical procedures developed here can simulate the mechanical behaviour of the spinal motion segments properly. PMID- 8718948 TI - Fit to diastolic arterial pressure by third-order lumped model yields unreliable estimates of arterial compliance. AB - The pressure pulse contour analysis method uses a third-order lumped model to evaluate the elastic properties of the arterial system and their modifications with adaptive responses or disease. A fundamental assumption underlying this method is that the estimates of model parameters (two compliances, an inertance, and a peripheral resistance) obtained from a measurement of cardiac output, and a simultaneous measurement of an arterial pressure, are independent of the pressure measurement site. If true, this hypothesis would provide a minimally invasive method for estimation of arterial compliance. The aim of the present study was to test the validity of this assumption and the ability of the method to assess changes of compliance in response to vasoactive drug administration. In five anaesthetised, open-chest dogs we measured pulsatile pressure and flow in the ascending aorta and pulsatile pressure in the terminal aorta, under basal, vasoconstricted (methoxamine), and vasodilated (sodium nitroprusside) conditions. Model peripheral resistance was assumed equal to the ratio of mean pressure to cardiac output. Estimates of inertance and compliances, and the associated estimation errors, were determined by fitting the model output to either the diastolic portions of ascending aortic pressure, P(adt), or terminal aortic pressure, Ptd(t). Results showed that the assumption of independency of model parameter estimates on the arterial pressure measurement site was not verified. Different images of the vasoactive drug-induced changes in vascular compliance were obtained from fits to P(adt) and Ptd(t). Model parameter estimates were associated with high estimation errors and were very sensitive to the choice of the period of diastolic pressure to be fitted. Model predicted aortic pressure, over the entire heart cycle, did not compare well with experimental ascending aortic pressure. Our results question the reliability of the pressure pulse contour analysis method for evaluating arterial compliance. PMID- 8718949 TI - Discussion on the design of a hip joint simulator. AB - Hip joint simulators were developed for predicting, by attempting to duplicate in vitro physiological loads and motion, the wear rate that total hip replacements are likely to show in vivo. From a theoretical point of view, loading and motion cycles of hip joints could be closely reproduced by three rotation actuators and three force actuators. However existing devices have been designed assuming that some of these degrees of freedom are negligible, in order to reduce the complexity of the equipment. The present study singles out some preliminary indications on the design choices regarding the spatial configuration of loading and motion actuators. The aim is to define theoretically a simplified simulator but still able to apply the most physiologically realistic loading cycle to the specimen. PMID- 8718950 TI - Finite element stress analysis of a hybrid fracture fixation plate. AB - Metal plates are commonly used in the operative treatment of bone fractures. Rigid metal plates stabilize the fracture site, maintain good contact between bone fragments and allow early weight bearing and patient mobility. However, treatment with rigid metal plates can cause localized bone atrophy due to stress shielding and interference with blood circulation, and the weakened bone can refracture after plate removal. A hybrid bone plate system that combines the torsional and bending rigidity of a metal plate with the axial compliance of a polymer insert has been designed. A three-dimensional, quarter-symmetric finite element model was generated for a canine femur diaphysis plated with this metal/polymer hybrid design. A model with a standard metal fixation plate was also generated for comparison purposes. The stress state in the underlying bone was examined for several loading conditions taken from published in vivo studies. The finite element model was used to study the performance of biodegradable polymer inserts in the plate system. The flexible plate reduced stress-shielding effects at the fracture site when subjected to an axial load. The bending strength of the plate was not compromised by the addition of the polymer inserts. Biodegradable inserts further enhanced the performance of the new plate design, transferring less of the axial load to the plate as the inserts broke down. PMID- 8718951 TI - A numerical model of an intensive care ventilator-humidifier system. AB - Current intensive care ventilator-humidifier systems neither monitor nor adequately control inspired gas humidity. Problems of low delivered humidity and condensation within ventilator circuitry are commonly encountered. To help to address these problems, a numerical model of a complete ventilator-humidifier patient intensive care system has been developed. The model, based on a finite difference technique, can predict pressures, flow-rates, temperatures and relative humidities at discrete points throughout the system. A comparison of numerical predictions and measurements in a real system is reported. A strong qualitative agreement is demonstrated in all cases studied, and a good quantitative agreement is obtained in most cases. It is concluded that such models could be used to assess methods of controlling ventilator-humidifier systems to prevent the occurrence of condensation. Similar models could be developed for other medical gas delivery systems. PMID- 8718952 TI - Reminiscences of John Romano and reflections on the state of psychiatric education. PMID- 8718953 TI - A different kind of education for psychiatric residents. PMID- 8718954 TI - Facilitating effective residency education on short-term inpatient units. AB - This paper examines the impact of shorter inpatient lengths of stay on residency education, and adaptations in training and supervision that facilitate effective learning experiences on a short-term inpatient unit. The caseloads of three PGY II psychiatric residents at the University of Rochester's Strong Memorial Hospital during three months of their inpatient rotations from 1979-80, 1986-87, and 1994-1995 are examined. Findings illustrate the changes occurring as inpatient units have moved to a shorter length of stay. The challenges and opportunities that face residents and supervisors in short-term inpatient settings today are examined. The discussion focuses on potential adaptations in the supervision of residents and organization of training experiences to insure that educational objectives are attained as length of stay decreases. Educational objectives especially relevant to short-term inpatient units are reviewed. Opportunities for continuity of teaching and training experiences offered by combining inpatient and partial hospital programs are highlighted. PMID- 8718955 TI - Recruitment and workforce issues in late 20th century American psychiatry. PMID- 8718956 TI - Less is more: the case for "basic" psychiatry and the Colorado Medical Student Log. AB - Because of the rapid growth of our field, psychiatric educators are faced with two related questions. Are we teaching our students about too many things thereby making it difficult for them to discern our essential message(s)? And, two, have we really decided on what our essential message(s) actually is? The answer is "yes" to the former and "no" to the latter. The problem is deceptively simple: as distinct from what we teach, as a profession we have not yet parsed out the minimum we expect our students to really know-and really know how to do-by the time they graduate from our schools. We, therefore, developed the menu and method of the Colorado Medical Student Log which articulates the minimal educational goals for our students: ten core clinical experiences and ten core psychiatric screens. The log also provides an easy method by which we can be satisfied that our minimal goals are being met. PMID- 8718957 TI - Psychiatry clerkship directors: who they are, what they do, and what they think. AB - We mailed questionnaires to ascertain biographic, academic and attitudinal data about psychiatry clerkship directors in the U.S. The average director is 46 years old, has been in academic medicine for 13 years, and has directed the clerkship for 5.9 years. Most are associate or full professors and hold one or more additional administrative positions, most often director of medical student education in psychiatry. Most view their role as fulfilling and want to direct the clerkship for the rest of their careers. However, most also perceive that their medical school does not provide enough support, that faculty don't teach enough, and that the current economic climate impedes learning. Unless medical student education is supported under managed care, the quality of clerkships and job satisfaction for clerkship directors will decline. PMID- 8718958 TI - Women's short-term recovery from cardiac surgery. AB - Cardiac surgery is becoming more prevalent in women. Previous knowledge and understanding regarding the process of recovery following cardiac surgery have been based predominantly on what has been known about men. Knowledge is needed regarding factors which may be particular to women's recovery from cardiac surgery and which include both biophysical and psychosocial components. A longitudinal study was conducted to describe, from both objective and subjective perspectives, women's short-term recovery from cardiac surgery. Preoperative (baseline) data were collected from 31 women who presented for cardiac surgery at two Northern California hospitals. Following discharge from the hospital, 27 women were followed monthly by telephone for three months to obtain subjective responses regarding postoperative symptoms, perceptions of recovery, activity, and health status. The findings indicate that women's perceptions of recovery are independent of NYHA functional classification, and these perceptions improve before other more objective measures of activity or health state demonstrate improvement. PMID- 8718959 TI - Patient and family adjustment to heart transplantation. AB - Heart transplantation is an effective treatment for end-stage cardiac disease and a major stressor to patients and their families. The purpose of this paper is to review the literature on patient and family adjustment to heart transplantation during the stages from pre-transplantation to post-transplantation. Most authors indicate that patients and families adjust well to heart transplantation. Support groups are a positive resource for heart transplant patients and their families. PMID- 8718961 TI - Antiarrhythmic drug therapy--Part I: General principles of drug selection. PMID- 8718960 TI - Hopelessness in the myocardial infarction patient. AB - Events that cause hopelessness following a myocardial infarction may include coronary heart disease, reality-based misperceptions, lack of future goals and lack of relatedness to others. The susceptibility to hopelessness will depend on how successful the patient has been in accepting previous cardiac-related problems. Once hopelessness occurs, the patient may perceive the future to be limited. Hopelessness centers around seven components: physiological, cognitive, temporal, affective, behavioral, affiliative and contextual loss of hope. This article outlines how hopeless patients may distort their immediate environment and future goals. Methods of identifying hopelessness and intervening with appropriate nursing interventions, which will assist with the management of hopelessness, are described. PMID- 8718962 TI - The beneficial effects of HMG-coenzyme A reductase inhibitor in the primary and secondary prevention of atherosclerosis in the coronaries. PMID- 8718963 TI - Refusing to pay for health care--Part II (of III): Emerging trends in third-party payer liability. PMID- 8718964 TI - In vitro antimicrobial efficacy of silver iontophoretic catheter. AB - A silver iontophoretic catheter was designed consisting of two silver wires connected to an electric power source and disposed in a parallel and helical manner around the proximal subcutaneous segment of a silicone catheter. In an in vitro tunnelled bridge model the silver iontophoretic catheter prevented the migration of Staphylococcus epidermidis from the highly contaminated hub to the sterile tip over a 40-d period. The silver impregnated cuff and electrically charged wires made of aluminium or iron delayed migration for only 72 h. A modified Kirby-Bauer technique, used to test the inhibitory activity of antimicrobial catheters, showed that the silver iontophoretic catheter has a broad spectrum inhibitory activity against Gram-positive bacteria, Gram-negative bacteria and Candida albicans. The silver iontophoretic catheter provides a long term electrochemical barrier against the migration of organisms from the external contaminated environment into the sterile intravascular compartment. PMID- 8718965 TI - 5-Fluorouracil release from copolymeric hydrogels of itaconic acid monoester. I. Acrylamide-co-monomethyl itaconate. AB - The aim of this work was to test the application of new copolymeric poly (acrylamide-co-monomethyl itaconate) (A/MMI) hydrogels to 5-fluorouracil (5-FU) release. Three different compositions of copolymers have been studied, 90A:10MMI 75A:25MMI and 60A:40MMI. The equilibrium swelling degree in saline solution was between 76 and 80% depending on the copolymer composition. 5-FU, as the sodium salt, was trapped in the gels by including it in the feed mixture of polymerization. The swelling kinetics of the hydrogels in saline solution were studied at four temperatures, and the diffusion coefficient and the activation energy of the process were obtained. The 5-FU release as a function of temperature and disc load was studied; the diffusion coefficient and the activation energy of the release process were also obtained. The diffusion studies follow Fick's second law. PMID- 8718966 TI - Alginate polycation microcapsules. II. Some functional properties. AB - The main cause of alginate polycation capsule breakage under physiological conditions is probably the osmotic swelling of the alginate core owing to the Donnan equilibrium set up by the negative charges of the carboxyl groups not involved in cooperative binding of counterions in the junction zones of the network. In the present paper we show how capsules can be stabilized extensively by reducing their swelling capacity in various ways. Alginate polycation capsules with good chemical and mechanical stability have been made by controlling their swelling behaviour through selection of capsule material according to chemical structure and molecular weight, as well as by controlling the kinetics of the capsule formation. Stable capsules have been made either by increasing the strength of the polyanion-polycation membrane, or by keeping a low-swelling gel network in the core. The latter capsules are made from an alginate rich in guluronic acid both in the core and in an outer coating, and with anisotropic distribution of the polymer material in the core where the concentration at the surface is higher than that in the centre of the capsule. Some functional properties of these capsules, such as porosity, have also been studied. PMID- 8718967 TI - Use of a biodegradable elastin-fibrin material, Neuroplast, as a dural substitute. AB - Since the new Neuroplast biomaterial is now commonly used in several domains of restorative and curative surgery, its preliminary evaluation as a dura mater substitute in the rabbit was performed and is described herein. Whereas the importance of prosthesis thickness was implicated and solved (0.2 rather than 1.0 mm), both macroscopic examination and histological studies revealed for the neotissue an effective patency, a good continuity and especially no or almost no adherence at all. As Neuroplast is now available for use everywhere in humans, this material is being developed in several fields of neurosurgery. PMID- 8718968 TI - Use of the glutaraldehyde-chitosan-treated porcine pericardium as a pericardial substitute. AB - The efficacy of chitosan post-treatment of glutaraldehyde-treated porcine pericardial substitute for the prevention of postoperative epicardial reaction and adhesion formation in mongrel dogs has been assessed. Glutaraldehyde (0.625%) treated porcine pericardium showed moderate to dense adhesions to the heart and other underlying organs with moderate to severe epicardial reaction. None to minimal epicardial reaction without adhesion was observed in glutaraldehyde treated porcine pericardium post-treated with chitosan at 5 months. Presumably, chitosan post-treatment prevents the slow release of residual glutaraldehyde and reduces the toxicity of the glutaraldehyde-treated implants, minimizing the chance of adhesion formation and epicardial reaction. PMID- 8718969 TI - Quantitative analysis of cell proliferation and orientation on substrata with uniform parallel surface micro-grooves. AB - In order to quantify the effect of the substrata surface topography on cellular behaviour, planar and micro-textured silicon substrata were produced and made suitable for cell culture by radio frequency glow discharge treatment. These substrata possessed parallel surface grooves with a groove and ridge width of 2.0 (SilD02), 5.0 (SilD05) and 10 microns (SilD10). Groove depth was approximately 0.5 micron. Rat dermal fibroblasts (RDFs) were cultured on these substrata and a tissue culture polystyrene control surface for 1, 2, 3, 5 and 7 days. After incubation the cell proliferation was quantified with a Coulter Counter, and RDF size, shape and orientation with digital image analysis. Cell counts proved that neither the presence of the surface grooves nor the dimension of these grooves had an effect on the cell proliferation. However, RDFs on SilD02, and to a lesser extent on SilD05 substrata, were elongated and aligned parallel to the surface grooves. Orientation of the RDFs on SilD10 substrata proved to be almost comparable to the SilD00 substrata. Finally, it was observed that the cells on the micro-textured substrata were capable of spanning the surface grooves. PMID- 8718970 TI - Characterization of a hyaluronic acid-Arg-Gly-Asp peptide cell attachment matrix. AB - We have developed a method to modify cross-linked hyaluronic acid with peptides containing the Arg-Gly-Asp sequence. The material created by this process is a three-dimensional porous matrix capable of supporting integrin receptor-mediated cell attachment. Peptide density can be controlled by varying the reaction conditions during peptide immobilization. Following cell attachment, cells actively proliferate and colonize the pores of the matrix. This material should prove useful for the maintenance of cells on a chemically defined three dimensional substrate or as a scaffold for enhancing tissue repair. PMID- 8718971 TI - Photopolymerized multifunctional (meth)acrylates as model polymers for dental applications. AB - Polymer networks that can serve as model systems for dental applications were prepared by photopolymerizations of 1,1,1-trimethylolpropane triacrylate, 1,1,1 trimethylolpropane trimethacrylate, 1,1,1-trimethylolethane trimethacrylate, ethylene glycol dimethacrylate, triethylene glycol dimethacrylate, Photomer 2028 and Photomer 3015. The UV polymerizations were initiated by 2,2-dimethoxy-2 phenyl-acetophenone. Volume shrinkage was followed over the course of polymerization using a dilatometric technique. Incident light intensities ranged from 1 mW cm-2 to 20 mW cm-2. The effects of monomer structure on % volume shrinkage, including pendant group size, molecular weight between reactive double bonds, and acrylate versus methacrylate monomers were investigated. In addition, the effect of incident light intensity on % volume shrinkage was studied. Typical volume shrinkage varied from 3.5% to 13.5%. The volume shrinkage decreased with increasing monomer rank and increased pendant group size; the shrinkage for methacrylates was less than that for acrylates. Increased incident light intensity resulted in increased shrinkage rate, but not in statistically significant increases of the volume shrinkage. Conversion was calculated from shrinkage data and compared to data from monomer extraction experiments. Results indicate that although double bond conversion is low, conversion of monomer units is significantly higher. PMID- 8718972 TI - Response of oral mucosal cells to glass ionomer cements. AB - Although glass ionomer cements are generally considered to be tissue-compatible, it has been suggested that unreacted components or setting reaction by-products can affect cell metabolism. The current study examined the effects of constituents leached out of three glass ionomer cements on growth and metabolism of oral epithelial cells. Aseptically prepared discs of Ketac-Cem Radiopaque (KCR), Ketac-Cem Maxicap (KCM) and Fuji I were incubated in Dulbecco's medium for 10 d, with daily medium changes. Cultures of hamster cheek pouch (HCP) cells, a line of hamster buccal pouch epithelial cells, were incubated in control or eluate-containing media for 24 h. Viable cell numbers were determined by the colorimetric MTS assay, and DNA and RNA syntheses were assessed using [3H]thymidine and [3H]uridine incorporation, respectively. Responses to materials were determined by comparison of cell numbers and radioisotope incorporation (counts per minute (cpm) per 1000 cells). Results were analysed by ANOVA and Duncan's multiple range test, then converted to percent control for comparison. The eluates of all three materials from the first 24 h of soaking inhibited HCP cell growth. The number of cells in cultures exposed to Fuji were 88% of control cultures, while those exposed to KCR and KCM were 58% and 59% of control, respectively. The difference between Fuji-exposed and control cultures was significant (P < 0.05). The two Ketac cements were different from Fuji-exposed and control cultures (P < 0.05) but not from each other. All of the materials caused significant increases in labelling of DNA compared to control cultures (P < 0.05) when calculated on a per cell basis, but the materials did not differ from each other. Both Ketac cements also significantly stimulated labelling of RNA per cell compared to control cultures (P < 0.05). All effects of the material decreased over time. Results suggest that leachable components of the materials may affect the rate of progression of HCP cells through the cell cycle, rather than overt toxicity that results in cell death. PMID- 8718974 TI - Comparison of bone regeneration with the use of mineralized and demineralized freeze-dried bone allografts: a histological and histochemical study in man. AB - Mineralized (FDBA) and demineralized freeze-dried bone allografts (DFDBA) have been proposed as substitutes for autologous bone in oral surgery. The demineralization process has been shown, in rodents, to determine osteoinduction in tissues other than bone. Other investigators have reported poor clinical results, in man, with the use of DFDBA. The aim of the present study was a comparative light microscopical and histochemical analysis of bone regeneration processes, in man, with the use of FDBA and DFDBA. Our histological results showed that in DFDBA only the particles near the host bone were involved in the mineralization processes, while in FDBA even the particles that were farthest from the host bone were lined by osteoblasts, actively secreting osteoid matrix and newly formed bone. These results probably point to a more osteoconductive effect of FDBA. No osteoinduction was observed with FDBA or DFDBA. PMID- 8718973 TI - Adhesion and proliferation of cultured human aortic smooth muscle cells on polystyrene implanted with N+, F+ and Ar+ ions: correlation with polymer surface polarity and carbonization. AB - Physicochemical surface properties and biocompatibility were studied in polystyrene (PS) implanted with 150 keV N+, F+ and Ar+ at doses ranging from 1 x 10(12) to 1 x 10(15) cm-2. Adhesion and proliferation of cultured human aortic smooth muscle cells (SMCs) on ion implanted PS were thoroughly examined for dependence on implanted dose and ion species and in close relation to polymer surface oxidation, surface polarity, concentration of conjugated double bonds and sheet resistivity. The surface polarity of PS was a smooth, increasing function of the implanted dose. However, the dependence of SMC population density on the implanted dose was found to be more complicated. After 18 h cultivation time (i.e. when only cell attachment and spreading took place), the number of adhered SMCs and their degree of spreading first increased with increasing ion dose, and after reaching a maximum at the dose of 5 x 10(12) cm-2, they decreased to original values. For doses above 5 x 10(14) cm-2, an increase in SMC population density and spreading was again observed. The first maximum in cell adhesion can be explained by slight increases in the surface polarity and wettability, optimal for cell adhesion, and the second maximum by progressive carbonization of the PS surface. After 96 h cultivation time (i.e. when the cells proliferated intensively), the dramatic dependence of the SMC population density on implanted dose is mostly smeared out. This observed dependence of SMC attachment, spreading and subsequent proliferation on the implanted dose was similar in all three ion species, but highest cell densities were achieved on PS implanted with F+ ions. PMID- 8718975 TI - Prevascularized bone graft cultured in sintered porous beta-Ca2P2O7 with 5 wt% Na4P2O7.10H2O addition ceramic chamber. AB - Autogenous bone transfer is an important part of reconstructive plastic surgery. Presently available techniques have the disadvantages of limitation of available donor site, loss of donor tissue and the possibility of donor defect or deformity. In the present study, a vascularized bone graft was created and cultured in the groin area of the New Zealand rabbit. The cylindrical ceramic chambers, 15 mm in length, 6 mm in outer diameter and 3 mm in inner diameter, were prepared by the addition of sintered porous beta-Ca2P2O7 with 5 wt% Na4P2O7.10H2O. In the first group, the chambers impregnated with autogenous bone fragments and allogenous demineralized bone matrix with volume ratio 1:1 were cultured in the rabbit's groin area with saphenous vessels passing through. In the second group, the chambers were treated by the same procedures as the first group but without saphenous vessels passing through. In the third group, the chambers were not impregnated, and were cultured in the groin area with saphenous vessels. After 2, 4, 6, 8 and 12 wk of operation, the animals were killed with an overdose of intravenous pentobarbital. The viability of the osseous tissue in the chamber was evaluated by histological examination, microangiograms and fluorochrome incorporation for the three groups. The autogenous bone chips could survive and retain their osteogenic properties while packed into the sintered porous beta-Ca2P2O7 (with 5 wt% Na4P2O7.10H2O addition) ceramic chamber and implanted in the rabbit groin area up to 12wk. However, even at the longest time periods, considerable amounts of dead bone were present in the chambers. In addition, we observed bone resorption in the three groups up to 12 wk, which might be attributed to lack of physiological stress. There were significant differences in new bone formation and osseous cell viability among the three groups. The prevascularized vessels and autogenous bone chips were both necessary for the formation of new bone and osteogenic property in the chamber under these heterotopic circumstances. The biodegradable ceramic used in this study was gradually absorbed and dissolved in the physiological environment. However, the degradation debris of the ceramic caused no injury to the new bone formation. These findings support the concept of creating a preformed vascularized bone graft to reconstruct segmental bone defects. PMID- 8718976 TI - Significance of interstitial bone ingrowth under load-bearing conditions: a comparison between solid and porous implant materials. AB - Interstitial bone ingrowth is extremely important for optimum fixation of implanted materials under load-bearing conditions. In this study, three types of biomaterial test piece were manufactured in solid and open-pore structures, and implanted into dog femoral condyles. Bone formation and remodelling were observed histologically and roentgenologically for 24 weeks thereafter. The study demonstrated that, 24 weeks after implantation, thick fibrous tissue surrounded by corticalized bone formed around both solid smooth-surfaced alumina and titanium implants. On the other hand, however, with an implant made of an artificial osteochondral composite material, thickening of ingrown trabeculae could be observed as early as 4 weeks. Bone ingrowth into the titanium fibre mesh was ambundant and increased with time after implantation. This interstitial bone ingrowth resulted in the complete integration of this implant and the viable host bone. Our findings suggest that interstitial bone ingrowth has great significance, even though new bone formation and remodelling follows Wolff's law after the completion of the bonding between the bone and implanted material under load-bearing conditions. The artificial osteochondral composite material could lead to complete integration of the implant and viable bone, suggesting that it is a promising material for joint replacements. Moreover, the tibial joint surface which bore against the polyvinyl alcohol hydrogel surface of this implant remained intact, which suggests that this composite is a very promising biomaterial for use in joint prostheses. PMID- 8718977 TI - Postoperative irradiation treatment and bioresorbable implants in orthopaedic surgery: an experimental in vitro study. AB - The influence of applied radiation on the degradation of a polyglycolide (PGA) test specimen was studied in vitro. There was no significant difference in the time-dependent degradation behaviour between the irradiated test specimen and controls. Mandatory irradiation of an operation site following surgery therefore does not contradict the employment of bioresorbable implants in orthopaedic surgery. PMID- 8718978 TI - Diagnostic clues from the surface ECG to identify idiopathic (fascicular) ventricular tachycardia: correlation with electrophysiologic findings. AB - INTRODUCTION: An RS interval > 100 msec in precordial leads has been recently described for the diagnosis of ventricular tachycardia (VT). The aim of this study was to assess the value of this criterion when applied to patients with right bundle branch block pattern, left-axis deviation (fascicular) VT sensitive to verapamil. METHODS AND RESULTS: Eleven patients (mean age 31 +/- 11 years; range 16 to 51) had a mean heart rate of 164 +/- 37 beats/min (range 107 to 230) during VT. The QRS complex axis was -92 degrees +/- -15 degrees (range -80 to 115). The mean QRS duration was 121 +/- 9 msec (range 105 to 140). The mean RS interval was 67 +/- 9 msec (range 60 to 80). Fusion beats were present in 2 patients (18%), and AV dissociation confirmed by electrophysiologic study was found on ECG in 8 (73%) of 11. During tachycardia, the QRS-H' interval was 19 +/- 10 msec (range 10 to 30) in 6 of 11 patients. In seven patients, a fast, unique (or double) presystolic potential lasting 32 msec (range 12 to 40) occurring before the onset of the QRS complex was found at the site of origin of VT, localized in the inferior apical left ventricular septum. In all cases, VT was successfully treated by catheter ablation. CONCLUSION: A wide QRS complex tachycardia with right bundle branch block and left-axis deviation sensitive to verapamil observed in a young patient without structural heart disease should not be confused with supraventricular tachycardia with aberrancy but rather suggests the presence of fascicular VT. As opposed to VT associated with structural heart disease, the RS interval is < 80 msec in all precordial leads in all cases. Independent of this parameter, AV dissociation detectable on surface ECG has a sensitivity of 73%, which increases to 82% in the presence of fusion beats. PMID- 8718979 TI - Effect of sustained load on dispersion of ventricular repolarization and conduction time in the isolated intact rabbit heart. AB - INTRODUCTION: It is well known that myocardial stretch can elicit ventricular arrhythmias in experimental models. However, previous reports have predominantly documented stretch-induced arrhythmias during short, pulsatile stretch. The arrhythmogenic mechanism of sustained static stretch is incompletely understood. METHODS AND RESULTS: To examine the influence of sustained load on several electrophysiologic parameters, a latex balloon was placed into the left ventricle of ten isolated Langendorff-perfused rabbit hearts and filled with a neutral volume of fluid. The heart was paced from a catheter inside the right ventricle (apicoseptal endocardial position), and the following parameters were studied during steady-state pacing with a cycle length of 500 msec (S1) and during extrastimulation (S2, base drive of 8 beats): monophasic action potential (MAP) durations at 90% repolarization (APD90) from 5 to 6 epicardial electrodes located on both ventricles and one right ventricular endocardial contact electrode; dispersion of APD90 (range of MAP durations from all electrodes); effective refractory period (ERP) and longest activation time (pacing stimulus to MAP upstroke). After baseline recordings, the balloon inside the left ventricle was filled with a volume of 1.0 mL of fluid by means of a servo-controlled pump. The ERP was significantly shortened from 198 +/- 9 msec at baseline to 183 +/- 8 msec during sustained load (P < 0.03). Similarly, the average APD90 was shortened from 180 +/- 5 msec at baseline to 175 +/- 6 msec during sustained load (P < 0.006) with steady-state pacing and from 178 +/- 6 msec to 170 +/- 8 msec during premature extrastimulation (P < 0.03). At the same time, dispersion of APD90 was increased from 27 +/- 5 msec to 38 +/- 6 msec (P < 0.002) during steady-state pacing and from 28 +/- 4 msec to 38 +/- 6 msec (P = 0.013) during premature extrastimulation. The longest activation time among all MAP recordings was increased from 39 +/- 2 msec to 43 +/- 3 msec (P = 0.003) during steady-state pacing and from 56 +/- 6 msec to 69 +/- 6 msec during premature extrastimulation (P < 0.003). CONCLUSIONS: Sustained load shortens the ERP and the mean APD90, and at the same time increases dispersion of APD90 and prolongs activation times. These findings provide additional insight into the arrhythmogenic mechanisms of sustained mechanical load. PMID- 8718980 TI - Mechanoelectrical feedback: maturation of a concept. PMID- 8718982 TI - Junction potentials and action potentials of the crayfish myocardium. AB - INTRODUCTION: The purpose of this investigation was to study the properties of junction and action potentials elicited by nerve stimulation in the absence of the cardiac ganglion in crayfish myocardium. METHODS AND RESULTS: The cardiac ganglion was surgically removed in isolated crayfish hearts. Electrical stimulation of one of the peripheral anterolateral nerves provided isolated junction potentials and action potentials free of the usual postspike bursts of junction potentials. The single junction potentials displayed amplitudes of up to 25 mV and slow exponential decay; the mean time constant was 170 +/- 13 msec (approximately x +/- SD). In fully recovered tissue, the junction potentials triggered action potentials free of repetitive subthreshold discharges. Tetrodotoxin did not alter the amplitude or shape of action potentials initiated by direct electrical stimulation of the muscle cells. Calcium channel blocking agents such as Cd2+ and Ni2+ eliminated the action potentials but not the junction potentials. Tetraethylammonium markedly prolonged the action potential duration. CONCLUSIONS: Our data suggest that: (1) A slow decay of the junction potentials may result from the disappearance of the neurotransmitter; this process also accounts for the late slow repolarization of the final part of the action potentials; (2) The equilibrium potential of the junction potentials is close to 0 mV; (3) The upstroke of the action potentials is carried by calcium currents; (4) The fast repolarization phase of the action potentials is likely caused by the delayed rectifier; and (5) The refractory phase outlasts the duration of the action potential. PMID- 8718981 TI - Altered pharmacologic responsiveness of reduced L-type calcium currents in myocytes surviving in the infarcted heart. AB - The pharmacologic responses of macroscopic L-type calcium channel currents to the dihydropyridine agonist, Bay K 8644, and beta-adrenergic receptor stimulation by isoproterenol were studied in myocytes enzymatically dissociated from the epicardial border zone of the arrhythmic 5-day infarcted canine heart (IZs). Calcium currents were recorded at 36 degrees to 37 degrees C using the whole cell, patch clamp method and elicited by applying step depolarizations from a holding potential of -40 mV to various test potentials for 250-msec duration at 8 second intervals. A Cs+ -rich and 10 mM EGTA-containing pipette solution and a Na+ -and K+ -free external solutions were used to isolate calcium currents from other contaminating currents. During control, peak ICa,L density was found to be significantly less in IZs (4.0 +/- 1.1 pA/pF) than in myocytes dispersed from the epicardium of the normal noninfarcted heart (NZs; 6.5 +/- 1.8 pA/pF). Bay K 8644 (1 micro M) significantly increased peak ICa,L density 3.5-fold above control levels in both NZs (to 22.5 +/- 6.2 pA/pF; n = 7) and IZs (to 12.8 +/- 3.0 pA/pF; n = 5), yet peak ICa,L density in the presence of drug was significantly less in IZs than NZs. The effects of Bay K 8644 on kinetics of current decay and steady state inactivation relations of peak ICa,L were similar in the two cell types. In contrast, the response of peak L-type current density to isoproterenol (1 micro M) was significantly diminished in IZs compared to NZs regardless of whether Ba2+ or Ca2+ ions carried the current. Thus, these results indicate an altered responsiveness to beta-adrenergic stimulation in cells that survive in the infarcted heart. Furthermore, application of forskolin (1 micro M and 10 micro M) or intracellular cAMP (200 micro M), agents known to act downstream of the beta receptor, also produced a smaller increase in peak IBa density in IZs versus NZs, suggesting that multiple defects exist in the beta-adrenergic signaling pathway of IZs. In conclusion, these studies illustrate that reduced macroscopic calcium currents of cells in the infarcted heart exhibit an altered pharmacologic profile that has important implications in the development of drugs for the diseased heart. PMID- 8718983 TI - Bundle branch reentry tachycardia and possible sustained interfascicular reentry tachycardia with a shared unusual induction pattern. AB - A case of bundle branch reentry tachycardia with an unusual induction pattern is presented. Unlike typical cases of this arrhythmia in which tachycardia is usually inducible with routine programmed ventricular stimulation and/or short long sequences, tachycardia in this case was inducible only with atrial stimulation. It also arose spontaneously during atrial flutter and during isoproterenol administration. After ablation of the right bundle, possible interfascicular reentry tachycardia with a similar induction pattern was observed. This tachycardia was successfully ablated in the region of the posterior fascicle of the left bundle branch. PMID- 8718984 TI - Transient T wave abnormalities after cessation of ventricular preexcitation: memory of what? AB - (1) Ventricular preexcitation causes both secondary and primary T wave changes. The secondary changes disappear immediately after cessation of preexcitation (ablation of accessory pathways) and unmask the primary T wave changes that regress within days, weeks, or months. (2) The pattern of primary T wave change produced by ventricular preexcitation depends on the location of the accessory pathway connections. The septal and posterior connections are associated with more prominent anteriorly directed T wave deflections and deviation of the T wave vector superiorly. The left lateral connections are associated with rightward deviation of the T wave vector in the frontal plane. (3) The primary T wave abnormalities are believed to be caused by local lengthening of repolarization, but the site of the postulated abnormalities needs to be established. (4) The persistence and slow dissipation of the primary T wave change induced by ventricular pacing, left bundle branch block, and presumably ventricular preexcitation have been attributed to the memory of past events. The nature of these events is not known. Several in vitro models demonstrated phenomena of gradual adjustment of ventricular action potential duration to change in cycle length, or to altered pattern of stimulation, but none of such models has mimicked the long-lasting regression of T wave abnormalities. (5) The terms T wave memory and pseudoprimary repolarization changes lack specificity and are unnecessary additions to the electrocardiographic vocabulary. PMID- 8718985 TI - Radiofrequency ablation of atrial flutter. AB - Activation mapping in common atrial flutter has shown circular (reentrant) activation of the right atrium around anatomic structures and areas of functional block. The direction of rotation is counterclockwise (in a frontal view), and in the low right atrium the myocardium between the inferior vena cava (IVC) and the tricuspid valve (TV) is critical to close the activation circle. The circuit can be interrupted by radiofrequency ablation of the myocardium between the TV and the IVC, and, in some cases, by ablation between the coronary sinus and TV. Flutter interruption does not mean complete isthmus ablation, as it may remain inducible, requiring further ablation. Despite attaining noninducibility, flutter may recur, and new procedures may be needed for complete ablation. Atrial fibrillation occurs in up to 30% of the cases during follow-up but is generally well controlled with antiarrhythmic drugs that were ineffective in treating flutter before ablation. Some noncommon atrial flutters show circular right atrial activation in a reversed (clockwise) direction, with the same critical areas in the low right atrium, and in these isthmus ablation is effective. Other noncommon flutters have different substrates in the right or left atrium, and mapping has to define specific critical isthmuses as ablation targets in each case. Left atrial flutter circuits remain inaccessible to ablation. PMID- 8718986 TI - Induced termination of fibrillation. AB - A previous communication in the Creative Musings section of this Journal summarized additions to the wavelet hypothesis related to the initiation of cardiac fibrillation. That hypothesis is also relevant to the termination of fibrillation, and further additions related to that event are presented in this report. Findings in both reports were obtained with a computer model based on the wavelet hypothesis, and results concerning initiation and termination of fibrillation were closely related. Refractory period (RP) conditions that terminated fibrillation were the inverse of those that increased vulnerability to the initiation of fibrillation. Increased RP range or decreased RP duration increased vulnerability to the initiation of fibrillation and decreased RP range or increased RP duration were capable of terminating fibrillation. Slow propagation increased vulnerability to initiation of fibrillation and acted to sustain fibrillation when instituted during fibrillation. The combination of increased duration and decreased range of RPs was more effective in terminating fibrillation than either alone. The magnitude of increased RP duration or decreased RP range required to terminate fibrillation and the effects of slow propagation on the maintenance of fibrillation depended on RP duration and range present during fibrillation. The findings extend the wavelet hypothesis of the nature of fibrillation to the prediction of conditions required to terminate fibrillation. PMID- 8718987 TI - What is the tachycardia mechanism? - III. PMID- 8718988 TI - Therapy of Wolff-Parkinson-White syndrome and concealed bypass tracts: Part I. PMID- 8718989 TI - Mental retardation in a birth cohort, 1976-1980, Rochester, Minnesota. AB - The cumulative incidence of mental retardation in a birth cohort of children born from 1976 through 1980 in Rochester, Minnesota (n = 5,919) was estimated. Rochester is the site of the Rochester Epidemiology Project, which captures virtually all medical care delivered locally. Passive follow-up through school and community medical records for criteria diagnostic of mild or severe mental retardation was undertaken. Thirty children were classified with mental retardation. The cumulative incidence of mental retardation by age 8 years was 9.1 per 1,000 (95% confidence interval = 6.2, 13.0) and was similar for boys and girls (8.3 vs. 10.0 per 1,000, respectively). The cumulative incidence for severe mental retardation in girls was more than twice that in boys; for mild retardation it was the opposite. The cumulative incidence among boys was 1.7 times greater than the cumulative incidence for girls. Results suggest that although the overall incidence of mental retardation by gender was similar, the severity may differ somewhat. PMID- 8718990 TI - Facial stereotypic movements and tardive dyskinesia in a mentally retarded population. AB - The facial stereotypies of adults diagnosed as having mental retardation and tardive dyskinesia were examined through a kinematic analysis of video-taped lip and tongue motions. A control group of healthy adult subjects without mental retardation was also examined in the production of preferred rates of lip and tongue oscillatory motions to provide a basis to assess the degree of movement variability in the stereotypies. The inter- and intraindividual variability of the movement form characteristics of the lip and tongue stereotypic motions was higher in the subjects with mental retardation. Results suggest that the low variability of discrete properties of movement kinematics may not be a defining feature of stereotypies. The concept of invariance in stereotypies may relate only to the topological kinematic properties of the movement sequence that provide the basis to infer that the same stereotypic movement sequence was reproduced from observation to observation. PMID- 8718991 TI - Immediate effects of mainstreamed settings on the social interactions and social integration of preschool children. AB - The immediate effects of mainstreamed and specialized settings on the peer interactions of preschool children with and without developmental delays were examined. Mainstreamed and specialized playgroups were established involving unacquainted peers and using a methodology that ensured appropriate matching of child and family characteristics. For each 2-week playgroup, the social and play interactions of each child were observed during a designated free-play period. Peer sociometric ratings also were obtained. Results indicated higher levels of peer interactions in mainstreamed settings for both typically developing children and children with developmental delays. The immediate impact of mainstreamed settings appeared to be attributed to the social demands and higher interaction levels of the former group. Children with developmental delays were not fully accepted nor totally socially integrated based on sociometric measures and behavioral indices of peer preferences. Implications of these findings for developing intervention programs to maximize children's peer-related social competence was discussed. PMID- 8718992 TI - Behavioral retardation in a macaque with autosomal trisomy and aging mother. AB - The social development of a female rhesus monkey (Macaca mulatta) was followed from the day of birth until her death, at age 32 months. The subject, born to an older mother, had an extra autosome (karyotype: 43, XX, +18), an affliction that came about spontaneously. MRI scans revealed that she was also hydrocephalic. Compared to 23 female monkeys growing up under identical conditions, the subject showed serious motor deficiencies, a dramatic delay in the development of social behavior, poorly established dominance relationships, and greater than usual dependency on mother and kin. The subject was well-integrated into the social group, however. PMID- 8718993 TI - Mapping the meanings of novel visual symbols by youth with moderate or severe mental retardation. AB - The word-learning ability of 12 school-age subjects with moderate or severe mental retardation was assessed. Subjects had little or no functional speech and used the System for Augmenting Language with visual-graphic symbols for communication. Their ability to fast map novel symbols revealed whether they possessed the novel name-nameless category (N3C) lexical operating principle. On first exposure, 7 subjects were able to map symbol meanings for novel objects. Follow-up assessments indicated that mappers retained comprehension of some of the novel words for up to delays of 15 days and generalized their knowledge to production. Ability to fast map reliably was related to symbol achievement status. Implications for understanding vocabulary acquisition by youth with mental retardation were discussed. PMID- 8718994 TI - Prevalence of dementia in adults with and without Down syndrome. AB - Declines in adaptive behavior were examined in a study of dementia in adults with Down syndrome and other forms of mental retardation. No significant differences were found between adults under 50 years of age with and without Down syndrome. In contrast, individuals over 50 who had Down syndrome were more likely to be classified as having dementia over a range of quantitative decision criteria; nevertheless, prevalence estimates of dementia were substantially below the presumed 100% prevalence of neuropathological markers of Alzheimer disease. This apparent discrepancy between functional and neuropathological findings may be associated with variations in risk associated with Down syndrome genotypes and/or a true lack of correspondence between classical neuropathological hallmarks of Alzheimer disease in this population and clinical expression. PMID- 8718995 TI - Plasma HVA in adults with mental retardation and stereotyped behavior: biochemical evidence for a dopamine deficiency model. PMID- 8718996 TI - Effortful and automatic processes associated with Down syndrome and nonspecific mental retardation. PMID- 8718997 TI - Arbitrary symbol learning by adults with severe mental retardation: comparison of lexigrams and printed words. PMID- 8718998 TI - [Climacteric--trigger for obesity?. Interview by U. Fuchs]. PMID- 8718999 TI - [Peripartal hysterectomy--definition, follow-up and risk]. AB - 39 peripartal hysterectomies from 1983 to 1993 were divided into three groups. Indication, post-operative development and complications were analysed. The first group contains the planned Caesarean section hysterectomies. The patients (n = 18) in this group hat the smallest blood loss and fewest complications. The second group includes the hysterectomies after Caesarean section without emergency (n = 8). The main reason was the rupture of the uterus without clinical signs during delivery. The complication rate of this group was also low. In the third group all operations were performed for vital risk of the patient (n = 13). Complications as placenta accreta (n = 8) or atonal bleeding of the uterus (n = 5) ended up in hysterectomy (eight after vaginal deliveries, five after Caesarean section). In this group we could observe a high blood loss as well as a high rate of complications. The prognosis of the patient depends on blood loss per time period, the dynamic of the bleeding. The mortality was higher in the group where patients needed a relaparotomy because of bleeding partially as a cause of clotting disorders. The most important task is to prevent shock situations by early sufficient blood transfusion. Because of the higher mortality of relaparatomy selective angiographic arterial embolisation should be considered. PMID- 8719000 TI - [Postoperative pain in patients after conventional abdominal and endoscopic surgery hysterectomy--a comparative study]. AB - The purpose of the present study is to determine differences in the postoperative character of pain between two study groups undergoing different techniques of hysterectomy. Therefore we examined 42 women. 21 women underwent combined laparoscopic and vaginal hysterectomy and 21 abdominal hysterectomies have been performed. All patients received on request Piritramid i.m. exclusively as an analgesic. The following factors have been included: 1. location of pain, 2. duration of pain, 3. maximum of pain, 4. analgesic requirements, 5. return to normal physical activities. With regard to our findings it seems to be certain that endoscopic operative hysterectomy compared to abdominal hysterectomy shows diminished duration of pain and reduction in the total amount of analgesic usage, as well as a shortened time to return to normal physical activities. PMID- 8719001 TI - [Supracervical uterus amputation via the vaginal route]. AB - A new method of supracervical amputation of the uterus (with or without extirpation of the adnexae) is presented. The operation is performed using a, the through vaginal route, opening the Douglas cavity through posterior coeliotomy. The advantages of the method, compared with Semm's laparoscopy operation, are discussed. PMID- 8719002 TI - [Gardnerella vaginalis: transport, microscopy, testing resistance]. AB - G. vaginalis is an important pathogen in the aetiology of bacterial vaginosis. Therefore, we investigated the influence of transport systems in isolation, a scoring system for Gram stains, and susceptibility to antimicrobial agents. The comparison between a simple (Transwab) and a sophisticated (Port-A-Cul) system showed no differences with regard to for instance Enterococcus faecalis or Escherichia coli; however, isolation of G. vaginalis, a fastidious microorganism, was significantly higher (alpha < 0.0001) in Port-A-Cul. There was a strong correlation (97.5%) using the scoring system indicating bacterial vaginosis and isolation of G. vaginalis. The minimal inhibitory concentrations (MIC) of metronidazole for 60 strains of G. vaginalis were higher than 32 mg/l, some strains showing heteroresistance. This phenomenon may be an explanation for treatment failures. Clindamycin and erythromycin were much more active, with MIC's between 0.016 and 0.19 mg/l, in-vitro development of resistance being slower for clindamycin than for erythromycin. CONCLUSIONS: (I) for isolation of G. vaginalis, a sophisticated transport system is mandatory; (II) a scoring system for Gram staining is helpful in diagnosis of bacterial vaginosis; (III) in patients with metronidazole treatment failures, clindamycin should be used. PMID- 8719003 TI - [Treatment and after-care of HIV patients with cervix dysplasia]. AB - We have been observing an increased prevalence of cervical intraepithelial neoplasia in HIV-infected women in our department of obstetrics and gynaecology. 10 HIV-infected patients with cervical intraepithelial neoplasia were treated by CO2-laser - 6 were subjected to laser excision conisation, and 4 patients to laser vaporisation. After laser vaporisation and laser excision conisation, no postoperative complications were observed. Intraoperative bleeding was rarely seen. The preoperative examination of the CD4-cell count had no influence on the choice of the applied method of laser treatment, laser vaporisation or laser excision conisation. But all the 4 patients with a CD4-cell count < or = 200/microliters experienced recurrence of disease shortly after the first laser treatment. Only one out of five patients with CD4-cell counts between 200 499/microliters had a recurrence of disease after laser treatment. All in all, a recurrence of disease was seen in five out of ten HIV-infected patients with cervical intraepithelial neoplasia. The recurrence rate is possibly increased because HIV-infected women have multifocal cervical, vulval and vaginal dysplasia more frequently than HIV-negative patients. In our study, we observed multifocal cervical, vulval and vaginal dysplasia in four HIV-infected patients. Therefore, an accurate short-term follow-up with colposcopy and cytological smears should be carried out after the treatment of HIV-infected patients with cervical intraepithelial neoplasia. PMID- 8719004 TI - [Labor, method of delivery and maternofetal transmission of HIV]. AB - Within a prospective study of the course of HIV-infection in women, 80 HIV infected women without AIDS were delivered of 80 children between 1985 and September 1992. The median of the age of gestation was 38 weeks. Until 1988 Caesarean section was chosen as mode of delivery (45 women). Later when the mode of delivery appeared to have no influence on the frequency of maternofetal HIV transmission, vaginal delivery was preferred (35 women). None of the infants was breastfed. Three infants --delivered vaginally--died within the first 6 months of life before their infection status could be determined. Seventy-seven children could be observed for 18 months or longer regularly every three months. 10 of the 77 children were found to be HIV- infected by serological, virological and clinical criteria. Taking into account the mode of delivery, of 32 children who were delivered vaginally or by emergency Caesarean section 8 were found to be HIV infected. None of 26 children delivered by elective Caesarean section after an uneventful pregnancy is infected. In 19 women Caesarean section was performed within 2 hours after onset of labour or after episodes of preterm labour which required hospital admission for treatment. Two children of these women are infected. No differences of CD4+ cell counts and p24 antigenaemia could be determined between the mothers of the three groups. The risk of fetal HIV infection was increased by preterm labour (p < 0.01) and the mode of delivery (p < 0.01). A correlation between loss of CD4 cells in the mother and increased risk of infection for the child is seen in children born spontaneously or delivered by emergency Caesarean section (p < 0.001). No correlation was found between the length of labour at delivery, the time of the rupture of membranes before birth as well as of the parity and the risk of fetal infection in that group. These findings point to labour as an important factor which increases the risk of maternofetal transmission of HIV. The onset of labour is accompanied by dramatic immunological alterations as a sudden increase of chemotactics and inflammatory cytokines at the maternofetal interface. The accumulation and stimulation of maternal immune cells will--if these cells are carriers of HIV--result in production and release of infectious HIV. This virus may accumulate in the maternofetal interface or gain access to the amniotic cavity. Since amniotic fluid is swallowed by the fetus, the largest potential port of entry for HIV in the fetus are the fetal lungs and the gastrointestinal tract. For the prevention of maternofetal transmission of HIV delivery before onset of labour or alternatively the protection of the fetus during parturition by means of potent antiviral compounds can be considered. PMID- 8719005 TI - [Termination of pregnancy in the 2nd trimester by serial administration of gemeprost vaginal suppositories. A prospective study (corrected)]. AB - In a prospective study termination of pregnancy was performed between the 16th and 24th week of gestation by serial applications of a 1 mg gemeprost containing vaginal pessary at 6h intervals. The reasons for termination were medical and fetal indications in 20 cases and missed abortion in 14 cases. The median induction to abortion interval was 17.4 hours (range: 3.7 - 73.0h) in the therapeutic abortion group, and a median of 3 applications were necessary to terminate pregnancy. In cases of missed abortion the median induction to abortion intervals was 11.0h (range: 3.5 - 32.5h) with a median of 2 necessary applications. Expulsion of the fetus occurred in 31 out of 34 patients (91.2%) within 24 hours. Spontaneous or surgically induced cervical lesions and blood loss > 500 ml were not observed, systemic prostaglandin-induced side effects were documented in 7 cases (20.6%). Epidural anesthetic gave a painless induction of abortion in all of patients. Our experience has shown that serial application of gemeprost vaginal pessaries is a feasible and efficient method for termination of second trimester pregnancies. PMID- 8719006 TI - [150 years Berlin Society of Obstetrics and Gynecology]. PMID- 8719007 TI - [Psychology of the desire for children in men and women]. PMID- 8719008 TI - [The 2nd legal abortion ruling of the German Legislative Court and its consequences]. PMID- 8719009 TI - [Future perspectives in contraception]. PMID- 8719010 TI - [Berlin Society for Obstetrics and Gynecology. 16 March 1994 Session]. PMID- 8719011 TI - [Serum hormones before and during therapy with cyproterone acetate and spironolactone in patients with androgenization]. AB - The effect of cyproterone acetate (CPA) and spironolactone (SPL) on the serum androgen concentrations of premenopausal women with symptoms of hyperandrogenism were investigated in a total of 39 women. The observation period was 12 months. CPA was administered according to the Hammerstein regimen: cyproterone acetate (CPA) [Androcur] 100 mg/die 5.-14. day of the cycle; ethinylestradiol (EE) [Progynon C]: 40 mg/die 5.-25. day of the cycle; Spironolactone (SPL) was given in a dosage of 100 mg/die from day 1.-21. of the cycle. During the therapy with CPA a significant decrease of total testosterone (61%), free testosterone (78%), LH (48%) and 17 alpha-Hydroxyprogesterone (72%) was observed; during the medication with spironolacton only a significant decrease of 5 alpha dihydrotestosterone (81%), which could not be seen during CPA use, was observed. Serum concentrations of total testosterone, free testosterone, LH and 17 alpha Hydroxyprogesterone remained unchanged. DHA and DHAS did not change during neither medication. Since peripheral androgens were not suppressed by SPL the positive therapeutical effect of SPL can be explained by the antiandrogenic effect at the level of the receptor. A disadvantage of spironolacton is the lack of contraceptive efficacy. In cases where contraindication for oral contraceptives are present SPL can be considered as a good alternative to CPA. The suppressive effect of CPA/EE on total testosterone, LH addition to the antivulatory effect makes it the preferable medication for hyperandrogenemic patients with polycystic changes of the ovaries (PCOD). PMID- 8719012 TI - [HELLP syndrome--amaurosis in sinus thrombosis with complete recovery]. AB - We report on a case of a 22-year old primigravida with HELLP syndrome in association with blindness and cerebral sinus thrombosis. The diagnosis of the cerebral sinus thrombosis was based on MRI 1h after delivery. Immediate caesarean section and intensive care treatment led to a complete recovery of the patient. PMID- 8719013 TI - [Spontaneous remission of HELLP syndrome in the 36th week of pregnancy with uterus duplex]. AB - We report on a case of an HELLP syndrome in the 36th week of gestation in a primigravida with known uterus duplex cum vagina duplice, which under intensive medical control showed a spontaneous decrease of the elevated liver enzymes and normalisation of platelet count. 12 days after the occurrence of the HELLP syndrome, a Caesarean section was performed because of PROM and breech presentation of the foetus. PMID- 8719014 TI - [Immunotherapy in gynecologic oncology]. PMID- 8719016 TI - Administration of glutamate following a reminder induces transient memory loss in day-old chicks. AB - Monosodium glutamate (4.0 mM) administered immediately after a visual reminder presented to day-old chickens between 7.5 min and 24 h following a single trial passive avoidance learning task produced transient losses of memory on retention test, an effect not observed in the absence of a reminder or when the reminder was given 48 h post-learning. The duration of the transient deficit decreased with increasing interval between the training and the reminder trial. The time of onset of memory loss after the reminder trial appeared to increase with increasing interval between the training and the reminder trials. The results suggest that, for a period of at least up to 24 h after passive avoidance training, retrieval of memory may lead to processes which are sensitive to inhibition by glutamate, with the duration of sensitivity post-retrieval decreasing as the period of memory consolidation increases. The results extend previously reported findings with rodents and suggest the possibility that consolidation of a stable memorial representation of a learning experience may take place over several days and may entail the concurrent laying down of a stable retrieval mechanism. PMID- 8719015 TI - [Ovarian suppression with Diane 35/50]. AB - In this study, hormone profiles were studied using Diane 35 and Diane 50 before and after treatment for 4-25 months. A comparison of the hormone profiles with Diane 35 or Diane 50 should investigate, if both preparations induce complete ovarian suppression, independent of the given dose of ethinyl oestradiol. 43 patients were treated with Diane 35 and 15 women with Diane 50 over a different lengths of time--up to a period of 25 cycles. Parallel to the determination of the hormone profiles (luteinising hormone [LH], follicle stimulating hormone [FSH], prolactin, oestrone [E1], oestradiol [E2], progesterone, total testosterone [T. T.], free testosterone [F. T.], dehydroepiandrosterone-sulfate [DHEA-S], delta-4-androstenedione [A.] and sex hormone binding globulin [SHBG]), observations on the androgen symptoms before treatment and their course during therapy were made. Moreover, the ratios of LH/FSH and E1/E2 as well as the androgenic index, the ratio of total testosterone/SHBG were determined as control of treatment effects. The most important hormonal changes, which had been observed during a period up to two years are the following: --LH and FSH suppression--prolactin with increasing tendency--decrease of the total androgens and free testosterone-- decrease of the oestrogens E1, E2--increase of SHBG. The confirmation of the efficacy of the anti-androgen/oestrogen combination was reflected by the decline of the androgenic index in all groups. Acne and seborrhoe improved after 3-4 cycles; alopecia after 8-9 cycles and hirsutism diminished after 3 cycles of treatment. In spite of different doses of the combined preparations with 2 mg cyproteronacetate and 0.035 or 0.050 mg ethinyloestradiol, no difference in ovarian suppression as well clinical effects could be found. PMID- 8719017 TI - Selective impairment of auditory attention following closed head injuries or right cerebrovascular accidents. AB - According to clinical experience a frequent consequence of head injury is an impairment of auditory attention. We investigated the possibility that patients with either closed head injuries (CHI), or cerebrovascular accidents (CVA) of the right hemisphere, would be impaired by comparison to healthy subjects on an objective test of auditory attention. We used an experimental paradigm that consisted of four subtests which comprised strings of auditory digits heard either diotically or dichotically, at either fast or slow presentation rates, respectively. Omission and commission errors were scored for each subtest and combined by an index of errors. The results showed that CHI patients were significantly impaired initially by the fast stimulus presentation conditions, whereas CVA patients made significantly more errors on the dichotic subtests independent of the speed of presentation. It is proposed that the observed selective attention deficits of these patients were due to differential disruptions of an interactive cortical network incorporating prefrontal, anterior cingulate, and temporoparietal structures of the right hemisphere. Statistically significant correlations between the error index scores and subjectively perceived attention deficits suggested that the auditory attention task measured clinically relevant aspects of attention. PMID- 8719018 TI - Modulation of event-related potentials by the repetition of drawings of novel objects. AB - In experiment 1, event-related potentials (ERPs) were recorded from 13 scalp sites while subjects viewed a series of line drawings of novel objects. Most of the drawings were of objects that were structurally possible, while the remainder were of structurally impossible objects; the task was to respond to each presentation of an impossible object. Approximately one third of the possible objects were repetitions of the immediately preceding drawing. Compared to the ERPs elicited by first presentations, the ERPs elicited by repetitions were more negative-going. This negative repetition effect comprised two temporally and topographically distinct components. In experiment 2, subjects monitored drawings of unstructured patterns, so as to detect occasional 'targets' containing a pair of parallel lines. Repetitions of 'non-target' patterns elicited ERPs which were largely indistinguishable from those elicited by first presentations. Thus, the negative repetition effects found in experiment 1 are not merely a consequence of repeating visual patterns in a demanding discrimination task. Possible reasons why novel objects should elicit ERP repetition effects opposite in polarity to those observed in tasks employing verbal or meaningful pictorial stimuli are discussed. PMID- 8719019 TI - Event related potentials recorded in Dorsal Simultanagnosia. AB - Visual evoked potentials (VEPs) to central and lateral half field patterned stimuli of 1, 2 and 4 cycles per degree (cpd) were recorded in a patient with Dorsal Simultanagnosia due to bilateral lesions of parieto-occipital junction. VEPs consisted of the normal N1-P1-N2 components with same spatial frequency sensitivity as in controls. VEPs had similar latencies and amplitudes whether the patient could see or not the patterned stimuli. Event related potentials (ERPs) to visual and acoustic odd-ball paradigm were also recorded in the same patient. Visual ERPs consisted of an early NA-effect, and of N2-P3 components. P3 was recorded only from frontal, central and temporal derivations. The topographical P3 abnormality was, however, the same for visual and acoustic odd-ball paradigms. The amplitude of P3 was smaller when the patient missed visual stimuli. These findings show that severe bilateral lesions at the parieto-occipital junction, inducing Simultanagnosia, do not obliterate VEPs or ERPs components. PMID- 8719020 TI - A new segmentation method of electroencephalograms by use of Akaike's information criterion. AB - Non-stationary EEGs, whose statistical properties change with time, were segmented into stationary segments to closely track the behavior of EEG characteristics. We have developed a new segmentation method of optimizing segmentation parameters by using AIC (Akaike's information criterion) as an objective criterion. We applied the segmentation method to EEGs. The instantaneous power spectra of EEGs estimated with wavelet transform were compared with the segmented EEGs. EEGs were recorded from F3, F4, C3, C4, P3, P4, 01 and 02 in 13 normal subjects. Artifact-free 15-s epochs were taken at each electrode location. Each epoch was divided into stationary segments, consisting of several fixed intervals, by optimizing 2 segmentation parameters (interval length and starting point) so that the sum of AICs for several sequences of segments could be the smallest. The EEG segmentation could represent differences in the power spectra between segments. The average length of segments during relaxed wakefulness was 6.0 +/- 3.8 s. The EEG segmentation during mental arithmetic could detect the start of mental arithmetic. PMID- 8719021 TI - Electrophysiologic correlates of age effects on visuospatial attention shift. AB - The effects of age on the neural processes underlying shifts in visuospatial attention were studied by non-invasive recording of brain electrical activity. Event-related evoked potentials (ERPs) were recorded in young (age 28-36 years old) and elderly (59-74 years old) subjects during a target detection task. Targets were preceded by central or peripheral informative cues, which are hypothesized to provoke voluntary and reflexive shifts of spatial attention, respectively. There were no age-related differences in the effects of cue validity on reaction times. Elderly subjects had longer reaction times to both the valid and invalid targets. The ERP recordings demonstrated that attention shift-related negativities (ARNs) were generated over the hemisphere contralateral to the central cue direction with the same amplitude in young and elderly subjects. However, the onset latency was delayed for the elderly at anterior scalp sites. Peripheral cues enhanced the N1 component (140-200 ms post cue) over the contralateral hemisphere comparably in the two age groups. The ARNs following N1 enhancement demonstrated delayed onset and restricted scalp distributions in the elderly group compared with the young group. These results suggest that resistance of cue validity effects to aging is consistent with existence of the N1 enhancement and ARNs, even in elderly subjects. Different age effects on ERPs generated by central and peripheral cues support the notion that voluntary and reflexive mechanisms of attention shift involve distinct neural systems with different vulnerability to aging. PMID- 8719022 TI - Reading habits and line bisection: a developmental approach. AB - 120 normal right-handed subjects, children and adults, with opposite reading habits (60 French, left-to-right readers, 60 Israeli, right-to-left readers) and 60 pre-school children (30 French and 30 Israeli), were submitted to a visuo motor bisection task. Bisection is found to be dependent upon reading habits with a leftward deviation of the subjective middle for left-to-right readers, and a rightward bias for right-to-left readers. Even before formal reading learning, French and Israeli pre-school children differ significantly in bisecting a line. Results are discussed with respect to hemispheric activation theories and directional hypotheses. PMID- 8719023 TI - The neuropsychology of depression and its implications for cognitive therapy. AB - The past few years have witnessed a growing interest in the specialty of neuropsychology as well as continued support for cognitive therapy of depression. The purpose of this paper is the examination of the neuropsychology of depression and its implications for A. T. Beck's cognitive theory and therapy of depression ([1963] "Thinking and Depression: Idiosyncratic Content and Cognitive Distortions," Archives of General Psychiatry, Vol. 9, pp. 324-333; [1964] "Thinking and Depression," Archives of General Psychiatry, Vol. 10, pp. 561-571; [1967] Depression: Clinical, Experimental, and Theoretical Aspects, New York: Harper & Row). Specifically, the neuropsychological and cognitive theory and therapy literatures related to depression are reviewed followed by an integration of these areas. Neuropsychological evidence is presented that both supports cognitive theory and therapy of depression and helps explain why such therapy may prove ineffective in treating depression. Implications for clinical practice, including neuropsychological assessment of depressives, and potential future research directions are also provided. PMID- 8719025 TI - Schizophrenia: a disease of heteromodal association cortex? AB - There is considerable evidence of disturbances of multiple brain areas in schizophrenia. The clinical features and findings from pathologic and neuro imaging studies suggest primary involvement of a system of parallel distributed networks within the neocortex--the phylogenetically recent heteromodal association cortex (HASC). There is evidence that HASC is a family of higher order parallel distributed networks of circuits, mediating complex representationally guided behaviors. We argue that HASC regions are especially involved in schizophrenia. Lesions of HASC in the disease are likely to be neurodevelopmental in origin (as evidenced by such examples as reversed planum temporale asymmetry) which have been identified by magnetic resonance imaging as specific regions of disproportionately reduced local gray matter volumes, and by neuropathologic examination as cellular migration disruptions. We believe the hypothesis of preferential heteromodal cortical abnormalities has heuristic value, and briefly indicate how it opens new avenues for investigating this debilitating condition. PMID- 8719024 TI - Detecting the malingering of cognitive deficits: an update. AB - There has recently been a dramatic increase of empirical studies that investigate methods for detecting malingering of cognitive deficits. The present review focuses on a comparison of simulated and suspected malingerers in the malingering literature, and critiques the numerous approaches to the detection of malingering. The approaches that are reviewed include detection of floor effects, discrepancies of information, response bias, neuropsychological tests and batteries, symptom validity testing, and anomalous performance on memory tests. The latter approach has only recently been proposed by researchers and may show the most promise. PMID- 8719026 TI - The 5-HT1A receptor: signaling, desensitization, and gene transcription. AB - The hypothesis that antianxiety or antidepressant agents (e.g., 5-HT1A agonists, 5-HT uptake blockers) exert their clinical actions via enhancement of serotonergic neurotransmission due to desensitization of 5-HT1A autoreceptors predicts that regulation of this receptor plays a crucial role in the therapeutic actions of these agents. A multidisciplinary strategy is described for the characterization of the 5-HT1A receptor at the level of cellular signaling mechanisms and genetic regulation, using heterologous expression of the cloned receptor in cell lines, site-directed mutagenesis, isolation of receptor-positive neuronal cell lines, and promoter analysis of the 5-HT1A receptor gene. These analyses will yield new insights into the possible mechanisms down-regulation of 5-HT1A receptor signaling, and may suggest novel sites of inherent defect involved in anxiety syndromes or major depression. PMID- 8719027 TI - Corticosterone alters 5-HT1A receptor-mediated hyperpolarization in area CA1 hippocampal pyramidal neurons. AB - The 5-hydroxytryptamine1A (5-HT1A) receptor in the CA1 region of the hippocampus is linked through a G protein to an inwardly rectifying potassium conductance. Activation of the 5-HT1A receptor results in a membrane hyperpolarization and decreases neuronal firing rate. The hippocampus contains a high concentration of the mineralocorticoid (MR) and glucocorticoid (GR) corticosterone (CT) receptor subtypes. Some laboratories have reported that CT modulates 5-HT1A receptor binding density and mRNA levels in area CA1 of the hippocampus; however, others have reported no change. Previous electrophysiological studies have demonstrated that acute (1 to 4 hour) MR activation in slices from adrenalectomized (ADX) rats attenuates the 5-HT1A receptor-mediated hyperpolarization, while acute MR+ GR or GR activation alone did not alter the 5-HT1A response. Our results confirm that the 5-HT1A response was attenuated 2 to 8 hours following MR activation. However, we found that GR activation alone decreased the potency, but not the maximal response to 5-HT. Chronic (2-week) treatment with basal levels of CT did not alter the 5-HT1A response. Administration of high concentrations of CT in vitro to neurons from chronically treated ADX rats decreased the magnitude of the 5 HT1A receptor-mediated hyperpolarization. We conclude that the 5-HT1A receptor effector system in CA1 hippocampal pyramidal neurons is modulated by CT in a dose and time-dependent manner. PMID- 8719028 TI - Cellular localization of the 5-HT1A receptor in primate brain neurons and glial cells. AB - Activation of 5HT1A receptors produces many different physiologic responses, which may be due to their localization on diverse cells in the brain. A 5-HT1A receptor antipeptide (aa170-186) antibody was produced that showed both high titer for peptide binding and immunocytochemical staining. Studies performed in perfusion-fixed brain tissue showed immunoreactive neurons, glial, and ependymal cells in the rat, mouse, cat, and monkey. Results from our studies of Macaca fascicularis brains are presented. We observed two main neuronal labeling patterns in the primate brain: (1) A general, diffuse somatodendritic distribution of 5-HT1A receptor immunoreactivity is seen in the raphe nuclei where the dendritic shaft, its branches and spines, and the entire perikaryon are immunolabeled. This pattern is also observed in the nucleus locus coeruleus, in scattered large brainstem reticular neurons, and in dentate gyrus hilar interneurons. (2) A discrete localization of 5-HT1A receptor immunoreactivity on the initial axon segment (axon hillock) is noted in pyramidal neurons of layer III and V of cerebral cortex, Cornu Ammonus (1-4) of the hippocampus, and in most brainstem and cervical spinal cord motoneurons. In addition to neuronal labeling, 5-HT1A receptor immunoreactivity is seen in the cell body and processes of astrocytes, and other nonneuronal cells. This pattern is particularly evident in the white matter of cerebral cortex and spinal cord, the pontine nuclei, the brainstem tectum, and the hilus of the dentate gyrus. The clinical implications of 5-HT1A cellular localization are briefly discussed. PMID- 8719029 TI - Is there a relationship between baseline and treatment-associated changes in [3H] IMI platelet binding and clinical response in major depression? AB - A peripheral model for the central 5-HT neuron is the characterization of platelet imipramine binding. We studied an outpatient major depressive cohort who fulfilled Research Diagnostic Criteria for agitation. After a 1-week placebo lead in, subjects were blindly randomized to either imipramine (IMI) or fluoxetine (FLU) during an 8-week, double-blind study period. Thirty-three subjects (15 IMI, 18 FLU) provided both baseline and endpoint samples for the platelet [3H]-IMI assay. Depression efficacy was comparable across the two treatments, whereas FLU was significantly more effective in reducing secondary anxiolysis (p = .023). Discontinuations due to an adverse event were significantly more frequent with IMI than FLU (p < .01). Baseline affinity (KD) was mildly predictive of change in the HAMD (r = -.22; p = .07). Whereas baseline to endpoint density (Bmax) changes (delta) were similar for IMI (183 +/- 329 fmol/mg) and FLU (196 +/- 402 fmol/mg), a statistically significant treatment difference in delta KD emerged (IMI -0.005 +/- 0.010 pmol/ml versus FLU 0.008 +/- 0.013 at p = 004). Moreover, the changes in KD and HAMD17 trended to a positive correlation among only the FLU-treated subjects (4 = 0.406, p = .095). The clinical effects of 5-HT-based selective antidepressant may be reflected by dynamic changes in the platelet 5-HT uptake apparatus. These data suggest that the baseline confirmational status of the [3H] IMI:5-HT transporter may reflect a "capacity" for a treatment response. PMID- 8719030 TI - Benzodiazepine receptor ligands with different intrinsic efficacies alter ethanol intake in alcohol-nonpreferring (NP) rats. AB - Benzodiazepine (BDZ) receptor ligands with varying intrinsic efficacies [RO19 4603, 0.02-0.15 mg/kg; FG 7142 1-16 mg/kg; DMCM, 1-8 mg/kg; RO16-6028 (bretazenil), 8-32 mg/kg] in modulating GABAergic activity were examined for the ability to alter palatability-induced ethanol (EtOH) intake in the alcohol nonpreferring (NP) line of rats. NP rats on a 22-hour fluid-deprivation schedule were given 2-hour daily access to a 10% (v/v) EtOH/3% (g/v) polycose solution and water. Average EtOH intake was 2.1 +/- 0.2 g/kg/2 hours, and water intake was 17.1 +/- 0.9 ml/2 hours. During the initial 15 minutes of the 2-hour session, RO19-4603, the imidazothienodiazepine partial inverse agonist reduced EtOH intake to 19% of control values at 0.04 mg/kg and completely suppressed drinking of the EtOH solution at 0.15 mg/kg. Twenty-four-hour postdrug administration, the 0.08 mg/kg dose of RO19-4603 completely suppressed drinking of the EtOH solution at the 60-minute interval, and the 0.15-mg/kg dose reduced intake to 20% of control levels at the 15-minute interval. FG 7142, the partial beta-carboline inverse agonist reduced EtOH drinking at the 60-minute interval with the 1-mg/kg dose, and the 16-mg/kg dose reduced water intake at the 15-minute interval. DMCM, the full beta-carboline inverse agonist, significantly reduced water intake at 15 minutes (4 and 8 mg/kg), and the same doses caused a substantial increase in EtOH drinking at the 120-minute interval. The anxiolytic agent bretazenil (16 and 32 mg/kg) increased EtOH consumption during the initial 15 minutes to 270% to 425% of control levels, and water intake increased by the end of the 2-hour session to as much as 210% of control following administration of the 32-mg/kg dose. These findings support existing evidence suggesting that BDZ receptor ligands may modify neuronal processes that mediate some reinforcing and/or aversive properties of alcohol. They further demonstrate a potential importance of the GABAA-BDZ receptor complex in mediating palatability- (environmentally) induced EtOH drinking even in rats selectively bred for low alcohol preference. PMID- 8719031 TI - Stability of interindividual differences in serotonin function and its relationship to severe aggression and competent social behavior in rhesus macaque females. AB - Few studies have investigated longitudinally interindividual stability of cerebrospinal fluid (CSF) 5-hydroxyindoleacetic acid (5-HIAA) concentrations in adult nonhuman primates across time and between baseline and stressful conditions. Furthermore, whereas studies with male macaques consistently have reported a significant, negative correlation between CSF 5-HIAA and rates of spontaneous aggression, wounding, and severe aggression, very few studies have examined this relationship in adult female nonhuman primates. Even fewer studies have investigated correlations between CSF 5-HIAA and competent social behavior, such as social dominance, in female monkeys. In the present study, two social groups of adult rhesus monkeys (Macaca mulatta) were formed by placing 16 females (aged 42 to 180 months, mean age: 68 months) in one of two indoor-outdoor enclosures with one or two adult males. CSF norepinephrine (NE), monoamine metabolites, and behavioral data were collected systematically over a 24-week period. In week 5 of the study, one additional female, not familiar to any of the other subjects, was added to each social group. Thereafter the groups were left undisturbed, and data characterizing spontaneous aggressive wounding and severe wound injuries in the females were collected for an additional year. The results showed that both group introduction and the addition of a new subject into each group resulted in increased monoamine turnover in the animals within the social groups. Interindividual differences in CSF concentrations of each of the monoamine metabolites and NE were highly stable from the baseline period to the stress samplings, and between stress samplings. Females with low CSF 5-HIAA exhibited higher rates of spontaneous aggressive wounding, and they were more likely to be removed from their social groups for aggressive wounding and/or treatment of injuries. CSF NE concentrations also were negatively correlated with rates of spontaneous aggression. In contrast, competitive aggression, i.e. noninjurious aggression used to maintain social dominance ranking, was not correlated with CSF 5-HIAA or NE. Females with above average CSF 5-HIAA prior to and following group formation were more likely to attain and maintain a high social dominance ranking within their social group than females with below average CSF 5-HIAA. The present findings indicate that CNS monoamine functioning in adult female rhesus macaques is traitlike, showing a high degree of interindividual stability across time and setting. These findings also suggest a role for serotonin in controlling impulses that regulate aggression and that competent social behavior among nonhuman primates may require average or above average serotonin functioning. PMID- 8719032 TI - Somatostatin. AB - 1. Somatostatin (SRIF) exerts diverse physiological actions in the body including regulation of hormone and neurotransmitter release and neuronal firing activity. Analogs of SRIF are used clinically to treat tumors and cancers and to block the hypersecretion of growth hormone in acromegaly. 2. The recent cloning of five SRIF receptor subtypes has allowed for the identification of the molecular basis of the cellular actions of SRIF. The ligand binding domains and regions involved in coupling to G proteins and cellular effector systems are being identified and the processes by which SRIF inhibits cell growth and proliferation are being established. Furthermore, subtype selective agonists have been generated which are being used to investigate the specific biological roles of each SRIF receptor subtypes. 3. Such information will be useful in developing a new generation of SRIF drugs that could be employed to treat metabolic diseases, disorders of the gut, cancer and abnormalities in the central nervous system such as epilepsy and Alzheimer's disease. PMID- 8719033 TI - Recent advances in molecular recognition and signal transduction of active peptides: receptors for opioid peptides. AB - 1. Opioid peptides are a family of structurally related neuromodulators which play a major role in the control of nociceptive pathways. These peptides act through membrane receptors of the nervous system, defined as mu, delta and kappa and endowed with overlapping but distinct pharmacological, anatomical and functional properties. 2. Recent cloning of an opioid receptor gene family has opened the way to the use of recombinant DNA technology at the receptor level. 3. This review focuses on the molecular cloning and functional characterization of opioid receptors and provides first insights into molecular aspects of opioid peptide recognition and signal transduction mechanisms, using the cloned receptors as investigation tools. PMID- 8719034 TI - Angiotensin II: biosynthesis, molecular recognition, and signal transduction. AB - 1. Angiotensin II is a well-known vasopressive octapeptide that is the principal end-product of the renin-angiotensin system. In addition to its tonic effect on vascular smooth muscle cells, it also stimulates aldosterone secretion from the adrenals and promotes sodium reabsorption through renal tubular cells. 2. These physiological functions have been appreciated for some time, but as details of the molecular and cell biology of the angiotensin response mechanism become understood, it is increasingly apparent that the hormone has a much broader repertoire. Its functional variability is made possible by (i) different enzymatic routes for its generation, (ii) different receptors distributed in different tissues, (iii) different mechanisms for receptor regulation, and (iv) different signal transduction pathways. 3. This insight is the direct consequence of advances in pharmacology that led first to inhibitors of angiotensin converting enzyme and later to angiotensin II receptor antagonists. This review looks at the current status of angiotensin biochemistry and physiology and provides a basis for anticipation of future developments. PMID- 8719036 TI - A VIP hybrid antagonist: from developmental neurobiology to clinical applications. AB - 1. The 28 amino acid vasoactive intestinal peptide, VIP, was originally isolated from the intestine, following a bioassay measuring vasodilating properties. Immunocytochemistry, receptor binding assays and in situ hybridizations have demonstrated VIP abundance in the nervous system, suggesting multiple bioactivities. 2. A pharmacological approach was chosen to dissect VIP activities and a prototype VIP antagonist (Met-Hybrid) consisting of a carboxyl fragment of VIP7-28 and a six amino acid fragment of neurotensin, neurotensin6-11-VIP7-28 was synthesized. 3. This hybrid peptide was designed to maintain the binding capacity of one parent molecule (VIP), while loosing the agonistic properties, representing a classical competitive receptor antagonist. Furthermore, the new molecule exhibited increased specificity to central nervous system VIP receptors. 4. The Met-Hybrid was originally discovered as a potent inhibitor of VIP function in vivo. In the adult rodent, acute administration of the antagonist resulted in blockade of VIP-mediated potentiation of sexual behavior and chronic intracerebroventricular application impaired VIP-associated learning abilities. During ontogeny, chronic injections of the molecule resulted in neuronal damage, disruption of the diurnal rhythmicity of motor behavior, and retardation in the acquisition of neonatal reflexes in the rat. 5. During gestation, severe microcephaly was induced by acute administration of the Met-Hybrid to pregnant mice. The hybrid antagonist inhibited VIP-stimulated mitosis in whole embryo cultures and in a variety of cancer cell lines in vitro and in vivo, suggesting therapeutical potential. PMID- 8719037 TI - Neurotensin receptors: binding properties, transduction pathways, and structure. AB - Neurotensin is a 13-amino acid peptide (pGlu-Leu-Tyr-Glu-Asn-Lys-Pro-Arg-Arg-Pro Tyr-Ile-Leu) originally isolated from hypothalami (Carraway and Leeman, 1973) and later from intestines (Kitabgi et al., 1976) of bovine. The peptide is present throughout the animal kingdom, suggesting its participation to important processes basic to animal life (Carraway et al., 1982). Neurotensin and its analogue neuromedin-N (Lys-Ile-Pro-Tyr-Ile-Leu) (Minamino et al., 1984) are synthesized by a common precursor in mammalian brain (Kislauskis et al., 1988) and intestine (Dobner et al., 1987). The central and peripheral distribution and effects of neurotensin have been extensively studied. In the brain, neurotensin is exclusively found in nerve cells, fibers, and terminals (Uhl et al., 1979), whereas the majority of peripheral neurotensin is found in the endocrine N-cells located in the intestinal mucosa (Orci et al., 1976; Helmstaedter et al., 1977). Central or peripheral injections of neurotensin produce completely different pharmacological effects (Table I) indicating that the peptide does not cross the blood-brain barrier. Many of the effects of centrally administered neurotensin are similar to those of neuroleptics or can be antagonized by simultaneous administration of TRH (Table I). The recently discovered nonpeptide antagonist SR 48692 (Gully et al., 1993) can inhibit several of the central and peripheral effects of neurotensin (Table I). Like many other neuropeptides, neurotensin is a messenger of intracellular communication working as a neurotransmitter or neuromodulator in the brain (Nemeroff et al., 1982) and as a local hormone in the periphery (Hirsch Fernstrom et al., 1980). Thus, several pharmacological, morphological, and neurochemical data suggest that one of the functions of neurotensin in the brain is to regulate dopamine neurotransmission along the nigrostriatal and mesolimbic pathways (Quirion, 1983; Kitabgi, 1989). On the other hand, the likely role of neurotensin as a parahormone in the gastrointestinal tract has been well documented (Rosell and Rokaeus, 1981; Kitabgi, 1982). Both central and peripheral modes of action of neurotensin imply as a first step the recognition of the peptide by a specific receptor located on the plasma membrane of the target cell. Formation of the neurotensin-receptor complex is then translated inside the cell by a change in the activity of an intracellular enzyme. This paper describes the binding and structural properties of neurotensin receptors as well as the signal transduction pathways that are activated by the peptide in various target tissues and cells. PMID- 8719038 TI - Ion channel hypothesis for Alzheimer amyloid peptide neurotoxicity. AB - 1. Alzheimer's disease (AD) is a chronic dementia and neurodegenerative disorder affecting the oldest portions of the population. Brains of AD patients accumulate large amount of the A beta P peptide in amyloid plaques. 2. The A beta P[1-40] peptide is derived by proteolytic processing from a much larger amyloid precursor protein (APP), and has been circumstantially identified as the toxic principle causing cell damage in the disease. 4. The A beta P[1-40] peptide is able to form quite characteristic calcium channels in planar lipid bilayers. These channels have conductances in the nS range, and can dissipate ion gradients quickly. The peptide can also cause equivalent cation conductances in cells. 5. We suggest that amyloid channel blocking agents might be therapeutically useful in Alzheimer's Disease, and have constructed molecular models of the channels to aid in the design of such compounds. PMID- 8719035 TI - Galanin--a neuropeptide with inhibitory actions. AB - 1. Galanin is a 29 (in humans 30) amino acids long neuropeptide with mostly inhibitory, hyperpolarizing actions. 2. Differential structural requirements of truncated forms of galanin and differential agonist/antagonist behaviour of chimeric peptides, high affinity galanin receptor ligands suggest the presence of pharmacologically distinct galanin receptor subtypes. 3. The galanin receptor from human Bowes melanoma cell line--a member of G-protein coupled receptor superfamily--has been cloned. 4. Galanin acts via Gi/G(o) proteins inhibiting cAMP production, inositol phosphate turnover, opening K+ channels or closing Ca2+ channels. PMID- 8719040 TI - Cholecystokinin receptors. PMID- 8719039 TI - Signal transduction of the gonadotropin releasing hormone (GnRH) receptor: cross talk of calcium, protein kinase C (PKC), and arachidonic acid. AB - 1. The decapeptide neurohormone gonadotropin releasing hormone (GnRH) is the first key hormone of the reproductive system. Produced in the hypothalamus, GnRH is released in a pulsatile manner into the hypophysial portal system to reach the anterior pituitary and stimulates the release and synthesis of the gonadotropin hormones LH and FSH. GnRH, a Ca2+ mobilizing ligand, binds to its respective binding protein, which is a member of the seven transmembrane domain receptor family and activates a G-protein (Gq). 2. The alpha subunit of Gq triggers enhanced phosphoinositide turnover and the elevation of multiple second messengers required for gonadotropin release and biosynthesis. 3. The messenger molecules IP3, diacylglycerol, Ca2+, protein kinase C, arachidonic acid and leukotriene C4 cross-talk in a complex networks of signaling, culminating in gonadotropin release and gene expression. PMID- 8719041 TI - Endothelin receptor heterogeneity, G-proteins, and signaling via cAMP and cGMP cascades. AB - 1. The endothelins (ETs) are potent vasoactive peptides which are involved in diverse biological processes, such as contraction, neuromodulation, and neurotransmission, as well as in certain pathophysiological conditions including cardiac and renal failure. 2. The diversity of action of ETs may be attributed to (i) the existence of a number of receptor subtypes, and (ii) the G-protein mediated activation of different signal transduction pathways. 3. The combined action of these two variables modulates the response, since different receptor subtypes can stimulate and/or inhibit the cAMP and cGMP cascades. PMID- 8719043 TI - Parental ancestry and risk of multiple sclerosis in Alberta, Canada. AB - Self-reported population ancestry data for the 19 census divisions (CDs) of Alberta, Canada, were correlated with multiple sclerosis (MS) prevalence rates in those divisions, for men and women separately; and parental ancestry was compared between a group of MS patients and controls attending the University of Alberta MS Clinic. At the CD level, there was a positive correlation between single Scandinavian ancestry and MS prevalence in men, but this was not confirmed in the case-control comparison. The case-control comparison indicated an excess risk of MS associated with single non-specific European as opposed to British ancestry in men only. When paternal versus maternal ancestry was considered separately, there was an excess risk of MS associated with non-specific European as opposed to British ancestry for both men and women, but on the father's side only. Aboriginal ancestry was negatively associated with MS prevalence at the CD level in both men and women; and no MS patients with aboriginal origin were among cases assembled through the MS clinic. PMID- 8719044 TI - Epidemiology of multiple sclerosis in Northwestern Sardinia: further evidence for higher frequency in Sardinians compared to other Italians. AB - The Sardinians are an ethnically homogeneous population, having a genetic structure quite different from that of all other Italian and European populations. All epidemiological studies carried out in Sardinia since 1975 indicate that this Mediterranean island shows twice the prevalence of multiple sclerosis (MS) compared to continental Italy, but the size of the Sardinian communities so far surveyed has been too small to draw definitive conclusions. To overcome this draw-back, we have studied the frequency of MS in a well-defined area of north-western Sardinia, with a population of about 270,000 in the 1991 census. Based on 276 MS cases, the prevalence on December 31st, 1991, was 102.6 per 100,000. The incidence, averaging 2 per 100,000 in the period of 1962 to 1971, rose to 5 in the period from 1977 to 1991. The present study confirms the higher frequency of MS among Sardinians compared to other Italian populations. Genetic, linguistic and historical data suggest a role of environmental and genetic factors in determining the notable difference in MS risk between Sardinia and the rest of Italy. PMID- 8719042 TI - Molecular neurobiology and pharmacology of the vasopressin/oxytocin receptor family. AB - 1. VP and OT mediate their wealth of effects via 4 receptor subtypes V1a, V1b, V2, and OT receptors. 2. We here review recent insights in the pharmacological properties, structure activity relationships, species differences in ligand specificity, expression patterns, and signal transduction of VP/OT receptor. 3. Furthermore, the existence of additional VP/OT receptor subtypes is discussed. PMID- 8719045 TI - Risk factors for dementia in Parkinson's disease: effect of education. AB - Cognitive deficits are common in Parkinson's disease (PD), but the pathophysiology and relationship to Alzheimer's disease (AD) are not understood. We used a case-control format to investigate putative risk factors for the development of dementia in patients with Parkinson's disease. We compared 52 cognitively intact patients with PD to 43 PD patients with dementia with regard to factors previously suggested as relevant to either AD or PD. Multiple logistic regression yielded the following significant predictors of dementia in PD: lack of education (less than a high school graduate) (OR 21); severity of motor deficit (UPDRS total motor score greater than 20; OR 6.34), and PD onset at greater than 60 years of age (OR 4.12). The predictive probability of dementia in our subjects when all three variables were positive was 97.9%. We conclude that education may modify the risk of cognitive decline in PD. Protective effects of educational attainment, independent of dementia etiology, may be due to greater functional brain reserve. PMID- 8719046 TI - Analysis of mortality in pergolide-treated patients with Parkinson's disease. AB - Parkinson's disease, because of its progressive degenerative nature, is associated with increased disability and mortality compared with mortality in the general population. We examined mortality data from three clinical trials involving 1,330 patients with Parkinson's disease treated with pergolide as an adjunct to levodopa or levodopa/carbidopa therapy. The ratio of observed deaths to expected deaths in the general population of the same age, gender, race distribution, and period of observation was 2.3 for the 3 studies combined. The ratio is lower than that in Parkinson's disease patients treated prior to the introduction of levodopa, consistent with ratios with levodopa and levodopa combination therapy. The ratio is slightly higher than in Parkinson's disease patients treated with levodopa and levodopa combination therapy, which may be attributable to differing patient characteristics in the populations studied. PMID- 8719047 TI - Prevalence of dementia in a semi-rural population of Catalunya, Spain. AB - This study was to investigate the prevalence of dementia in an aging population. A two-phase model was used to obtain information on the socio-demographic, medical and cognitive status of subjects over 65 years of age (n = 516), resident on December 31, 1990, within the general population (n = 3,457) of La Selva del Camp. A diagnostic protocol, following the criteria of DSM-III, was designed for application to all subjects. We diagnosed 64 subjects with dementia, which represented a prevalence of 14.9% of which 3.2% was classified as severe, 4.5% as moderate and 7.3% as slight. The prevalence by age and sex showed a large increase with age and a higher prevalence in females, although the latter was not statistically significant. PMID- 8719048 TI - Cognitive stability over 2 years in a rural elderly population: the MoVIES project. AB - In an ongoing prospective community study, a random sample of rural elderly persons was screened with cognitive tests (including the CERAD neuropsychological battery) at study entry and an average of 2 years later. We examined 1,017 subjects, nondemented at study entry, at both waves, with the Mini-Mental State Exam, Story Recall, Word List Recall and Recognition, Boston Naming Test, Verbal Fluency, Praxis, Clock Drawing, and Trailmaking. Overall, the cognitive performance was stable, with either no mean change or a small mean decline over 2 years; however, standard deviations were relatively large, implying individual variation of questionable clinical significance. These data provide a set of population-based longitudinal cognitive norms and have implications for dementia screening. PMID- 8719049 TI - Interpretation of secular increases in incidence rates for primary brain cancer in Connecticut adults, 1965-1988. AB - Available summaries of radiology reports were examined among 899 primary brain cancer cases (age 40 years and older) diagnosed in Connecticut residents in selected years from 1965 to 1988 and reported to the population-based Connecticut Tumor Registry. Adjustment for the lower sensitivities of radiologic tests used before the advent of computerized tomography (CT) suggested that the introduction of CT (by itself) could account for little of the secular increase in brain cancer rates. Examination of trends in age-standardized rates for histologically confirmed brain cancers in the elderly, after excluding those diagnosed only by radiologic tests, did not support the idea that the secular trend in the elderly was largely artifactual. PMID- 8719050 TI - Medicine and the Canadian state: from the politics of conflict to the politics of accommodation? AB - This paper analyzes data from three large-scale surveys of Canadian physicians conducted over the past decade to examine the politics of a cohort of recently established family physicians in Ontario, and to assess the extent to which these politics represent a "softening" of professional resistance to government health insurance. Politically, this is an important cohort because the physicians in it have grown up without any firsthand knowledge of the pre-Medicare period, and because they are among the first to establish practices in the wake of the month long 1986 Ontario physicians' strike, a high point of profession-government conflict. Factors which may have contributed to a moderation of medical politics include the progressive entry of women into medicine. Our data suggest that professional opposition to Medicare is declining and that fewer physicians support a return to voluntary and commercial control of the health system, a shift which could assist in breaking the historical cycle of profession government conflict and moving to the politics of accommodation. In the conclusions we discuss implications for medical politics in Canada and other countries such as the United States. PMID- 8719051 TI - Delivering the goods? Access to family physician services in Canada: a comparison of 1985 and 1991. AB - In recent years considerable attention has been given to the effects of universal, first-dollar coverage for health care services as provided under Canada's health care system. However, no consideration has been given to the stability of these effects through periods of changing economic climate. In this paper we consider the extent to which the achievements of the Canadian approach to health care funding have been maintained in the presence of increasing attention to cost containment. Multivariate analyses are used to (a) explain variations in utilization of family physician services among the population and (b) explore the relationship between utilization and need for periods of differing economic circumstances. We observe that the relative importance of differences in need in explaining variations in use among the population was less in the period when cost containment was of greater concern. These findings indicate that policymakers cannot assume that removing financial barriers to access to service, although important in achieving a more equitable distribution of service utilization, may be sufficient to sustain such distributions during time of constraint. PMID- 8719052 TI - Organization, management, and client effects on staff burnout. AB - Employee burnout is perceived to be a serious problem in human service organizations serving persons with severe mental illness. There has been considerable investigation of burnout and many of its possible causes, but little investigation of the role of client severity in relationship to burnout. Furthermore, theoretical models linking environmental context, organization structure, and management processes to burnout have seldom been proposed or examined. Building on our previous work, we employ a theoretical framework that conceptualizes environmental context, organization structure, management processes, client severity and staff characteristics as influencing work satisfaction and burnout. We examine this in a survey of 311 staff in 42 community mental health service organizations. Controlling for individual staff characteristics, results suggest that organization structure, culture, and management process are important to work environment and in turn to satisfaction and subsequently to burnout. Contrary to the literature, client severity was not associated with burnout nor to work dissatisfaction. PMID- 8719053 TI - Welfare and psychological distress: a longitudinal study of African American urban mothers. AB - This paper examines the psychological impact of receiving welfare. The prospective longitudinal data are from a cohort of African American mothers (N = 833). Four explanations structured the analyses: social selection; chronic burdens; a failure hypothesis; and earlier life conditions of welfare recipients. Psychological distress was assessed by women's reports of depressed and anxious moods. Women were categorized into one of four welfare conditions by receipt of welfare when their children were first-graders and 10 years later. Concurrent and longitudinal relationships existed between the women's welfare status and reports of psychological distress. Welfare recipients had more chronic burdens--including perceived ill health--which helped to explain these relationships. Measures of failure did not "explain'' the greater psychological distress. Women who grew up receiving welfare were more likely to report psychological distress and lower self-esteem later in life. The women's education influenced welfare and physical and psychological well-being. PMID- 8719054 TI - Lifetime traumas and mental health: the significance of cumulative adversity. AB - This paper reports the community prevalence of 20 life traumas and considers their individual relevance as risk factors for psychiatric disorder. Also presented is the first evidence on the mental health significance of cumulative adversity as indexed by a count of lifetime exposure to a wide array of potentially traumatic events. The question of the importance of considering such events within efforts to assess variations in life stress is also examined. Our results demonstrate clear relationships between many traumatic events and, especially, accumulated lifetime trauma experience and both psychological distress and psychiatric disorder. That these relationships persist with temporal priority controlled - and net of the effects of parental psychopathology - suggest the causal relevance of major lifetime events and the conclusion that they represent on important dimension of increased mental health risk. From these findings and from evidence for the significance of traumas in disorder recurrence, it is contended that failure to take account of such events has resulted in the systematic underestimation of the role of stress exposure in accounting for variations in emotional distress and disorder. PMID- 8719055 TI - Family cohesion as a buffer against the effects of problem-drinking fathers on psychological distress, deviant behavior, and heavy drinking in adolescents. AB - In this paper we examine the degree to which family cohesion buffers the effects of fathers' problem drinking at Time 1 (T1) on adolescent distress, deviance, and heavy drinking at Time 2 (T2), one year later. Data from a representative sample of 658 families were used to test the hypotheses. Mothers, fathers (if present), and adolescent children were interviewed in the home. Fathers who were present completed self-report scales measuring problem drinking. When fathers were not available, mothers' reports on fathers' drinking were used to measure fathers' problem drinking. Results from regression analysis indicate that after controlling for the effects of race, SES, age, gender, and family structure: (1) the more cohesion in the family and the fever stressful events, the less distress, deviance, and heavy drinking shown by adolescents; (2) the fathers' problem drinking affects adolescent distress and deviance when cohesion is low; but as cohesion increases, the effects of the fathers' drinking are reduced. The findings support the hypothesis that cohesion in families buffers the effects of fathers' problem drinking on adolescents. PMID- 8719056 TI - The psychological consequences of multiple roles: the nonnormative case. AB - Occupying multiple roles has been shown to be a major predictor of psychological well-being. We investigated the effects of multiple roles in the nonnormative case: a sample of aging mothers of adult children with mental retardation. These women have been in the role of caregiver for up to five decades, in addition to occupying the range of roles normatively held in adulthood and old age. We found, using longitudinal analyses, that holding multiple roles in significantly and negatively related to depression in this sample, as in the general population. The effect of multiple roles remains significant even after the previous level of depression is controlled. These results support the role accumulation hypothesis predicting positive consequences of multiple roles on psychological well-being, even in this sample of older women who have had lifelong caregiving responsibilities. PMID- 8719057 TI - The search for maximal levels of reimbursement input for a given work output. PMID- 8719058 TI - Best practice and enterprise bargaining. AB - The Australian Council of Trade Unions (ACTU) and its affiliated health unions have strongly supported and actively participated in the Federal Government Best Practice in Health initiative over the past two years. We believe the initiative has been an important vehicle in fostering the development of the Best Practice approach in this vital sector of the economy. Healthcare expenditure in this country runs to some $34.3 billion (1992/93 figures) annually, financed by a mix of Commonwealth, State and private sources and constituting approximately 9% of GDP. Hospital expenditure alone absorbs just over a third of the total at $11.8 billion (1990/91). At the same time, we have developed one of the most accessible and equitable healthcare systems in the world and an increasing source of valuable export dollars. The Australian trade union movement, like others in the community, recognizes that real cost constraints exist on governments in healthcare as elsewhere. Those constraints can lead to a variety of responses, such as to withdraw money from the system, combined with total reorganization of financing through the introduction of casemix and hospital reorganization, to see whether the system survives. The alternative is one epitomized by a Best Practice type approach. It is these types of approaches and their relationships with enterprise bargaining on which I intend to focus. However, I hasten to add that the issue of context is all important. Whilst it may be possible to create a microcosm of Best Practice change in a particular healthcare setting, it is unlikely to be maintained. The interface with the socio-ecomonic and political context cannot be avoided and it is this context which will determine the continuing success of Best Practice initiatives--whether in health or elsewhere. PMID- 8719059 TI - Mystery shopping as a quality adjunct in public health organizations. AB - Public health in Australia is undergoing unprecedented emphasis on meeting customers' needs, wants and expectations. Mystery shopping, common in the commercial world, has much to offer health organizations in their push towards quality. This paper describes this inexpensive technique and reports on its use in North West Health Service, a large rural health provider. The potential exists for its widescale adoption in health to better meet customer focus objectives. PMID- 8719060 TI - Validation of a 28 day interval between discharge and readmission for emergency readmission rates. AB - For the majority of studies on hospital readmission rates, a readmission is deemed to have occurred if a patient was admitted within 28 days of the previous discharge date. This time period, however, has rarely been justified clinically or statistically and certainly not in Australia. In examining the patterns of readmissions to NSW public acute hospitals, this study provides statistical support for the use of a 28 day interval for total readmissions. The same finding applies to the four admission categories of medical, surgical, paediatrics, and obstetrics and gynaecology. PMID- 8719061 TI - Limited adverse occurrence screening: an effective and efficient method of medical quality control. AB - The objective of the present study was to determine the rate of adverse patient occurrences in a medium sized hospital and to assess the effectiveness and efficiency of limited adverse occurrence screening as a method of medical quality control. The medical records of inpatients discharged from a base hospital in Horsham, Victoria, were screened by the medical records department using eight general outcome criteria. Histories found to meet a criterion were sent to a medical reviewer to determine if an adverse patient occurrence had taken place. A random sample of histories not meeting any criteria was also reviewed. The main outcome measures were the rate of adverse patient occurrences and the proportion of these events detected by limited occurrence screening, the accuracy of the screening process, the time taken and the cost of finding adverse events. The total adverse patient occurrence rate was estimated to be 2.75% (95% CI 1.36 4.14%). Limited adverse occurrence screening using eight screening criteria detected 49.1% (95% CI 32.6-99.3%) of all adverse patient occurrences and 64.4% (95% CI 37.8-100%) of all adverse occurrences of major severity. This was achieved by reviewing the records of 9.72% of all patients discharged. Screening was quick and accurate (false positive rate 2.0%, false negative rate 0.4%). Medical review took on average 5 min (s.d. +/- 3.03). The method required 500 h of staff time over one year and cost $22000 (0.1% of total hospital budget). The proportion of adverse patient occurrences found by limited screening was much higher than that found by traditional quality assurance methods. Limited adverse occurrence screening using retrospective review requires a small proportion of total budget that should be available to most hospitals for medical quality assurance activities. PMID- 8719062 TI - Audit of paediatric appendicectomy results. PMID- 8719063 TI - ACHS accredited health-care facilities at 4 October 1995. PMID- 8719064 TI - Differences in initial treatment patterns and outcomes of lung cancer in the elderly. AB - BACKGROUND: Non-small cell lung cancer (NSCLC) accounts for substantial deaths and costs in the elderly greater than 65 years old. The current practice of NSCLC treatment in a Medicare population was examined to ascertain important areas of practice variation, and differences in clinical outcome and costs. METHODS: Data from incident cases of NSCLC from the Virginia Cancer Registry (VCR), 1985-89, were matched with claims from Medicare Part A and B, census tract data and the Area Resource File. Multivariate models were created to include clinical data, demographics, and access information. RESULTS: For patients with locoregional disease, increasing age was associated with lower likelihood of therapy (odds ratio (OR) 0.35; confidence intervals (CI) 0.29, 0.43), thoracotomy (OR 0.27; CI 0.21, 0.34), and more use of radiation therapy compared to surgery (OR 1.69; CI 1.39, 2.03). Low education levels were associated with less likelihood of treatment (OR 0.78; CI 0.66, 0.94), or radiation instead of surgery (OR 1.22; CI 1.05, 1.47). Patients in urban areas were less likely to receive therapy (OR 0.67; CI 0.49, 0.92). For distant disease, increasing age was also associated with lower likelihood of treatment (OR 0.48; CI 0.41, 0.56), as was increasing co morbidity (OR 0.84; CI 0.75, 0.93). Distance to radiation oncologists made no difference in radiotherapy utilization. Two year survival according to therapy was surgery 66%, radiation 15%, no therapy 17%. CONCLUSIONS: Patterns of care, and survival according to therapy, vary widely for elderly NSCLC patients. Age, low education, higher co-morbidity and urban residence all decrease the likelihood of surgical therapy for locoregional NSCLC. Despite the availability of coverage through the Medicare program, use of therapies and survival is not uniform for all beneficiaries. Possible discrimination by age, co-morbid illnesses not recorded in the Medicare files, or patient and provider choice could all be involved; administrative billing files cannot resolve these important differences. PMID- 8719065 TI - Survival predictors in advanced non-small cell lung cancer. AB - The authors studied the influence on survival of 21 clinical, anatomical, haematological and biochemical factors evaluated, at diagnosis, of 411 patients (pts) with advanced Non Small Cell Lung Cancer (NSCLC) followed in our department between 1984 and 1990. Most of the patients were male (347--84.4%) and only 64 (15.6%) were females. Median age was 62 years, but was slightly higher in females. Only 34 patients were aged under 45 years. Squamous cell carcinoma (215 pts--52%) and adenocarcinoma (152 pts--37%) were the most frequent histologic types. Performance status was poor--only 103 (25%) continued active; 120 (29%) spent at least half of the time in bed; 188 (46%) were severely limited. After staging, 179 (44%) presented locally advanced disease (stage IIIB) and 232 (56%) metastatic dissemination (stage IV). Therapy was defined by the oncologic group according to individual characteristics and based on clinical grounds. Anti neoplastic therapy was performed in 225 (55%), chemotherapy alone in 121 (30%), radiation therapy alone in 67 (16%), and sequential combined treatment (chemotherapy and thoracic radiation) in 37 (9%). Until 1987, the main chemotherapy regimen was MACC (Metrotrexate + Adriamycine + Cyclophosphamide + Lomustin), afterwards VP(M) (Cisplatin + Vimblastin + Mitomycine). Radiation therapy was performed using Co60, 2 Gy/day, 5 days a week, for 4 weeks (approximately 45 Gy total). The response rate was poor--four complete responses (2%), 42 (19%) partial responses. The overall median survival was 4.3 months and only 5% of patients were alive after 18 months of follow up. Prognostic importance of each characteristic studied was initially done by unifactorial analysis, followed by multifactorial analysis according to two methods: Cox proportional hazards model and recursive partitioning amalgamation--RECPAM. Regardless of the method used, the main determinants of survival were found to be performance status (Zubrod), weight loss and serum albumin. Other factors such as the staging (presence or absence of metastasis), lymphocytes, lactic dehydrogenase, and hoarseness were also significant. It is noteworthy that age and histological type were irrelevant; sex and hoarseness only proved important when integrated within a multifactorial model. The overall prognostic evaluation and therapeutic decision of advanced NSCLC patients could be improved by combining the prognostic value of TNM with that of performance status, weight loss and serum albumin. These prognostic guidelines must be taken into account when designing new clinical trials. PMID- 8719066 TI - Long-term survival of surgical cases of lung cancer. AB - The survival rate and important prognostic factors were studied in patients observed for more than 10 years after surgery for squamous cell carcinoma and adenocarcinoma. The causes of death of long-term survivors and whether 5- or 10 year survival assures complete cure were also examined. The records of 324 patients operated on at Kyoto University Chest Disease Research Institute between January 1976 and May 1984 were analyzed. Five- and 10-year survival of these patients was 34.4 and 26.8%, respectively. There was no difference between the survival rate of adenocarcinoma and squamous cell carcinoma. Significant difference were noted in survival curves between stage I, II and IIIa, and stage IIIa and IIIb cancers (P = 0.001, P = 0.005, and P = 0.001, respectively). A significant difference in the survival curve was also observed between those with complete and with incomplete resection (P < 0.0001). Multivariate analysis using Cox proportional hazards method showed that ages, pathological N-factor, and complete resection were significant prognostic factors. Nine of the 16 patients who died between 5 and 10 years after surgery had recurrent or second primary lung cancer. Five of these nine patients were diagnosed as recurrence or distant metastases. Of the 39 patients who survived disease-free for 10 or more years after surgery, none died of new recurrence. It is speculated that 5-year survival would not indicate complete cure after surgery for lung cancer and patients should be followed up for more than 10 years. PMID- 8719067 TI - Clinical implication of symptoms in patients with non-small cell lung cancer treated with definitive radiation therapy. AB - To clarify clinical significance of symptoms presented at the beginning of treatment, we analyzed the symptoms in 240 patients with non-small cell lung cancer treated with definitive radiation therapy. Symptoms were classified into four groups: no symptom (Grade 0), cough, sputum/hemosputum and fever up (Grade 1), chest pain and breathlessness (Grade 2), appetite loss, body weight loss, SVC syndrome, hoarseness, and pain in the upper limb/shoulder (superior sulcus tumor) (Grade 3), and their therapeutic outcomes were examined. The 2- and 5-year overall actuarial survival rates for patients with squamous cell carcinoma were 38.5% and 15.4% for Grade 0, 40.5% and 20.1% for Grade 1, 17.9% and 2.6% for Grade 2, and 15.8% and 5.3% for Grade 3. A statistical difference was noted in survival between Grades 0-1 and Grades 2-3 (P < 0.01), but was not seen between Grades 0 and 1, or between Grades 2 and 3. As for patients with stage III disease alone, the difference in survival was still significant between Grades 0-1 and 2 3 (P < 0.05). In patients with adenocarcinoma/large cell carcinoma, however, no obvious relationship was found between symptoms and prognosis, except for body weight/appetite loss. In conclusion, though the symptoms closely related to clinical stage and performance status and not an independent prognostic factor, chest pain and breathlessness correlated with poor therapeutic outcome as well as body weight loss and T3-4 related symptoms, while cough, sputum (hemosputum), and fever were favorable symptoms in patients with squamous cell carcinoma who received definitive radiation therapy. PMID- 8719068 TI - Etoposide (VP-16) and cisplatin at maximum tolerated dose in non-small cell lung carcinoma: a Cancer and Leukemia Group B study. AB - A multi-institutional cooperative group trial was undertaken by the Cancer and Leukemia Group B (CALGB) to evaluate the efficacy of the combination of cisplatin and intravenous etoposide for the treatment of metastatic or recurrent non-small cell lung cancer (NSCLC). The doses used were those previously determined to be the maximally tolerated dose of this drug combination. Forty patients were entered into the trial, 37 of whom were eligible for evaluation. Cisplatin (35 mg/M2/day for 3 days) and etoposide (200 mg/M2/day for 3 days) were administered every 28 days for a planned 6 cycles of therapy. Sixteen of 37 evaluable patients (43%) responded to therapy. Myelosuppression was the dominant toxicity, with 89% of the patients experiencing grade 4 neutropenia, and nearly half grade 3 or 4 thrombocytopenia. Median survival was 8.5 months, with 30% of the patients alive at 1 year and 10% alive at 2 years. Malaise, fatigue, and peripheral neuropathy were the other major toxicities. The combination of etoposide at the dose of 200 mg/M2/day for 3 days and cisplatin at 35 mg/M2/day for 3 days is a highly potent combination against metastatic non-small cell carcinoma. PMID- 8719069 TI - Combination chemotherapy with vindesine-ifosfamide-cisplatin (VIP) in locally advanced unresectable stage III and in stage IV non-small cell lung cancer: a phase II trial. AB - The efficacy and toxicity of a regimen adding ifosfamide to the more classical cisplatin-vindesine combination was studied in patients with advanced non-small cell lung cancer. Sixty-four good performance patients with inoperable stage III or stage IV were treated with VIP: vindesine 3 mg/m2 days 1 and 8, ifosfamide 1200 mg/m2 and platinum 30 mg/m2 days 1, 2 and 3, repeated every 4 weeks, up to a maximum of six cycles. Response rate, clinical data and radiological tests were rigourously reviewed by a panel. Overall response rate was 39% (95% confidence interval, 27%-51%) with three patients achieving a complete response; response rate in stage III was 48%. Median survival was 9 months. Toxicity consisted mainly of bone marrow toxicity and nausea/vomiting, but was manageable. There was no renal toxicity greater than grade 2, four severe infections, but no treatment related deaths. CONCLUSION: VIP as mentioned above is very active in good performance patients with advanced non-small cell lung cancer. Its activity, together with its manageable toxicity--without severe renal or pulmonary toxicity -makes it an attractive candidate for induction chemotherapy. PMID- 8719071 TI - Well-differentiated fetal adenocarcinoma of lung. AB - Well-differentiated fetal adenocarcinoma (WDFA) histologically resembles pulmonary blastoma, and is thought to be a subtype of pulmonary blastoma which has differentiated epithelial features resembling the fetal lung among its epithelial features and sarcomatous features. We recently encountered one patient who underwent surgery for WDFA. This case is reported with a discussion of the literature. A 33-year-old woman had a tumor shadow in the lower lobe of the right lung. The tumor was diagnosed as pulmonary blastoma as a result of echographic biopsy, and right total pneumonectomy was performed. No sarcomatous features were observed on postoperative histological assessment, and the patient was diagnosed as having WDFA. Its prognosis is believed to tend to be better than that of biphasic blastoma, in which sarcomatous features are mingled with epithelial features. However, it is reported that chemotherapy or radiotherapy has seldom been effective. Complete surgical resection is essential for long-term survival. PMID- 8719070 TI - Subacute encephalopathic toxicity of cisplatin. AB - Acute blindness or seizures are usually the first signs of central neurotoxicity from cisplatin. We report a case of subacute neurotoxicity caused by cisplatin. Progressive encephalopathy and partial loss of vision were the main observed signs. This condition was completely reversible upon cisplatin discontinuation, as is usually the case with acute central neurotoxicity. PMID- 8719072 TI - A model of "sense of effort" during maximal and submaximal contractions of the tongue. AB - Fatigue in the oral motor system may be accompanied by the perception of an increased "sense of effort." Awareness of centrally generated motor commands that result in synaptic inputs to motoneuron pools are thought to be responsible for these perceptions of effort (e.g., Muller, 1840; McCloskey, 1981). Few studies of the perceptual phenomenon of sense of effort exist, particularly of the oral motor system. The present study required 20 normal adults to push on a fluid filled bulb using their tongue and hand. Subjects repeatedly exerted from 10 to 100% of the maximal effort, in random order, in 10% increments. The pressure produced by pushing on fluid-filled bulbs was compared to the effort level attempted. Subjects produced consistent, reliable pressures related to effort level. The best mathematical model for both the tongue and hand data was third order polynomial. It is hypothesized that the perception of effort derives from a central source that operates across various motor systems. PMID- 8719073 TI - Electrophysiologic manifestations of impaired temporal lobe auditory processing in verbal auditory agnosia. AB - The present study examined the extent to which verbal auditory agnosia (VAA) is primarily a phonemic decoding disorder, as contrasted to a more global defect in acoustic processing. Subjects were six young adults who presented with VAA in childhood and who, at the time of testing, showed varying degrees of residual auditory discrimination impairment. They were compared to a group of young adults with normal language development matched for age and gender. Cortical event related potentials (ERPs) were recorded to tones and to consonant-vowel stimuli presented in an "oddball" discrimination paradigm. In addition to cortical ERPs, auditory brainstem responses (ABRs) and middle latency responses (MLRs) were recorded. Cognitive and language assessments were obtained for the VAA subjects. ABRs and MLRs were normal. In comparison with the control group, the cortical ERPs of the VAA subjects showed a delay in the N1 component recorded over lateral temporal cortex both to tones and to speech sounds, despite an N1 of normal latency overlying the frontocentral region of the scalp. These electrophysiologic findings indicate a slowing of processing of both speech and nonspeech auditory stimuli and suggest that the locus of this abnormality is within the secondary auditory cortex in the lateral surface of the temporal lobes. PMID- 8719074 TI - Speech-language outcomes of hemispherectomy in children and young adults. AB - Children and young adults who had undergone right or left hemispherectomy for intractable seizures after a period of normal language acquisition were compared with respect to scores on speech and language tests. The majority of the subjects had full scale IQs in the borderline to mentally retarded range. Language scores were computed in relation to estimated mental age, not chronological age. On this basis, the left hemispherectomized children were more likely to show syntactic comprehension and rapid-rate auditory processing deficits than the right hemispherectomized. The two groups were similar to one another and to normal children in speech production. The findings are discussed in relation to developmental language disorders. PMID- 8719075 TI - Speech prosody in affective contexts in Thai patients with right hemisphere lesions. AB - The production of speech prosody in emotional contexts was examined in Thai patients with unilateral right hemisphere damage. Twelve right hemisphere patients and nine normals read target sentences embedded in paragraphs that cued either a happy, sad, or neutral affect. Perceptual evaluations of their productions revealed a severe deficit in right hemisphere patients. Acoustic analysis indicated that longterm measures of fundamental frequency, timing, and energy at the sentence level were aberrant in right hemisphere patients. Findings are discussed in relation to nature and extent of prosodic deficits in right hemisphere patients, hemispheric specialization for linguistic and affective prosody, and the effect of language structure on the manifestation of affective prosody. PMID- 8719076 TI - Developmental deep dyslexia in Japanese: a case study. AB - This report demonstrates the existence of developmental deep dyslexia involving Japanese orthography. When asked to read (or name) isolated kanji, T.S., a sixth grader with a normal IQ and no speech impairment, produced a number of visual, selection, and semantic errors. It is suggested that these errors arise from an interaction between phonological coding impairment and the relative sparing of direct visual processing, which are taken to be characteristic of developmental deep dyslexia. A selection hypothesis is presented to explain selection errors, semantic errors, and some peculiar phenomena observed in acquired dyslexics. PMID- 8719077 TI - The interpretation of sentence ambiguity in patients with unilateral focal brain surgery. AB - Interpretations of ambiguous sentences were studied in patients with unilateral anterior temporal lobectomy or selective amygdalo-hippocampectomy. The sentences represented lexical and syntactic ambiguities. In both left- and right-sided groups, regardless of type of surgery, total mean score on the test was below normal. Left-sided cases, regardless of type of surgery, provided significantly fewer alternative interpretations than right-sided cases. The results suggest greater left than right hemisphere specialization in both lexical and syntactic processing, but also suggest right hemisphere involvement in resolution of lexical ambiguity. PMID- 8719078 TI - Trace deletion, theta-roles, and cognitive strategies. AB - This paper reports a rather striking distinction found in the performance of agrammatic patients: Their comprehension deficit distinguishes not only different syntactic constructions, but also verbs with different thematic structure. Thus, the coupling of a variety of sentence types with different verb types yields surprising performance patterns, demonstrating once again that the deficit these patients suffer from is highly selective from a grammatical viewpoint. This pattern of selectivity has important implications for linguistic theory, for it describes an error pattern that can be accounted for only by a theory that assumes a (normal) lexicon that encodes thematic labels and refers to a thematic hierarchy. In addition, the results argue for the necessity of a cognitive strategy (first NP = Agent) as part of the structural account of agrammatic comprehension (the Trace-Deletion Hypothesis). It is shown that, contrary to some recent claims, an account lacking such a strategy cannot derive agrammatic performance patterns properly. New data are presented, from an experiment that used an anagram task to test agrammatic comprehension of active and passive sentences containing verbs of three different thematic types. Verbs varied in that their external argument was Agent, Instrument, or Experiencer. The finding was that on all active sentences, performance was above chance, whereas on the passives, performance split: on verbs with Agent or Instrument external arguments, performance was at chance, corroborating previous findings. On Experiencer verbs, though, performance was below chance. These data argue decisively for the TDH as originally presented, namely, for an account that assumes trace deletion, coupled with a strategy. The implications of these results to the theory of brain/language relations are discussed, in the context of the theory of lexical representation. PMID- 8719079 TI - Emergent control of manual and vocal-motor activity in relation to the development of speech. AB - Babbling typically precedes, resembles, and conceivably facilitates development of speech, and yet there is no accepted neurobiological characterization of babbling. Here we report a study of infants' developing control of vocal behavior in relation to manual activity performed under differing conditions of audibility. We hypothesized that babbling is associated with the onset of left lateralized motor control, as expressed in repetitive right-handed activity, and that audibility facilitates such activity. Sixty-one normally developing infants were seen before (N = 21) or at various intervals following (N = 40) the onset of babbling. In experimental trials, audible or inaudible rattles were placed in left or right hands equally often. Analysis of manual activity revealed little shaking movement in the youngest and vocally least differentiated infants, and a sharp increase in shaking in slightly older infants who had recently begun to babble. Surprisingly, audibility only marginally enhanced shaking activity. A dextral bias was evident in the shaking of infants who had recently begun to babble, but not in younger or older infants. These and other findings suggest that the left cerebral hemisphere may be disproportionately involved in the production of repetitive vocal-motor activity as occurs in babbling. PMID- 8719080 TI - An in vitro biomechanical study of a multiplanar circular external fixator applied to equine third metacarpal bones. AB - The biomechanical characteristics of a 4-ring circular multiplanar fixator applied to equine third metacarpal bones with a 5 mm mid-diaphyseal osteotomy gap were studied. Smooth Steinmann pins, either 1/8 inch, 3/16 inch, or 1/4 inch, were driven through pilot holes in the bone in a crossed configuration and full pin fashion and fastened to the fixator rings using cannulated fixation bolts. The third metacarpal bone fixator constructs were tested in three different modes (cranial-caudal four-point bending, axial compression, and torsion). Loads of 2,000 N were applied in bending and axial compression tests and a load of 50 N x m was applied during testing in torsion. Fixator stiffness was determined by the slope of the load displacement curves. Three constructs for each pin size were tested in each mode. Comparisons between axial stiffness, bending stiffness, and torsional stiffness for each of the three different pin sizes were made using one way analysis of variance. There was no visually apparent deformation or permanent damage to the fixator frame, and no third metacarpal bone failure in any of the tests. Plastic deformation occurred in the 1/8 inch pins during bending, compression, and torsion testing. The 3/16 inch and 1/4 inch pins elastically deformed in all testing modes. Mean (+/-SE) axial compressive stiffness for the 1/8 inch, 3/16 inch, and 1/4 inch pin fixator constructs was: 182 +/- 16 N/mm, 397 +/- 21 N/mm, and 566 +/- 8.7 N/mm; bending stiffness was 106 +/- 3.3 N/mm, 410 +/- 21 N/mm, and 548 +/- 12 N/mm; and torsional stiffness was 6.15 +/- 0.82 N x m/degree, 7.14 +/- 0.0 N x m/degree, and 11.9 +/- 1.0 N x m/degree respectively. For statically applied loads our results would indicate that a 4 ring fixator using two 1/4 inch pins per ring may not be stiff enough for repair of an unstable third metacarpal bone fracture in a 450 kg horse. PMID- 8719081 TI - Spinal cord swelling as a myelographic indicator of prognosis: a retrospective study in dogs with intervertebral disc disease and loss of deep pain perception. AB - The association of myelographic spinal cord swelling with neurological outcome was examined in 46 dogs with intervertebral disc disease and absence of deep pain perception (DPP). Spinal cord swelling was measured by calculating a ratio of the length of the loss of the myelographic dye column to the length of the second lumbar vertebra (L2). A positive neurological outcome was defined as return of voluntary motor function. A cut-off value for swelling:L2 of 5.0 was established by the creation of a receiver operator characteristic curve. Using a swelling:L2 ratio of 5.0 as a cutoff for indication of neurological recovery yielded a sensitivity of 74% and a specificity of 61%. Overall neurological recovery rate was 43%. Dogs with spinal cord swelling:L2 ratios less than 5.0 had a recovery rate of 61%, whereas dogs with a ratio greater than or equal to 5.0 had a recovery rate of 26%. Evaluation of these data by chi square analysis confirmed that a ratio less than 5.0 was associated with a positive outcome, and a ratio greater than or equal to 5.0 was associated with a negative outcome, (P < .05). Although other factors, such as duration of neurological signs, affect neurological outcome in dogs with no DPP, evaluation of myelographic spinal cord swelling can assist in establishing a prognosis. PMID- 8719082 TI - A mucosal apposition technique for ureteroneocystostomy after renal transplantation in cats. AB - Renal transplantation is a successful treatment for terminal renal failure in cats. However, in the initial clinical study, there was a frequent occurrence of obstruction of the implanted ureter at the bladder wall or stoma. This resulted in the use of a modified "drop-in" technique that had proved effective in the prevention of obstruction in five normal cats. When applied to renal transplant recipients, ureteral obstruction was reduced, but continued to occur. The modified "drop-in" technique was abandoned and replaced with a technique that apposed the cut edge of the ureteral mucosa to the torn edge of the bladder mucosa. This technique is recommended as it has prevented ureteral obstruction in six successive cases. Prevention of ureteral obstruction, and the required corrective surgery, markedly reduces patient morbidity and mortality, length of hospitalization, and expense to the client. PMID- 8719083 TI - Deep-frozen allogeneic cancellous bone grafts in 10 dogs: a case series. AB - Deep-frozen, aseptically collected and processed allogeneic cancellous bone was implanted in eight dogs during the surgical repair of diaphyseal long bone fractures and in two dogs during arthrodeses. A combined allogeneic and autogeneic cancellous bone graft was used in two fractures with a segmental bone loss of more than 5 cm. Bone union occurred in five fractures and in both arthrodeses. Failure of fixation occurred in two dogs with nonunion fractures and in a third dog with an open, infected fracture. Biopsies from the fracture sites were obtained from these dogs following failure of their fracture fixation. The cancellous bone graft appeared to be in the process of normal incorporation in each case. Failure of fixation was attributed to technical or case management errors or both, in each of the three fractures that failed to achieve bony union. Frozen allogeneic cancellous bone grafts were effectively incorporated when used in the primary repair of fractures and arthrodeses. Combined autogenous and allogeneic cancellous bone grafts may be particularly useful in the repair of fractures with large segmental diaphyseal bone defects. The use of allogeneic cancellous bone grafts in nonunion fractures requires further investigation before it can be recommended. PMID- 8719084 TI - Controlled cross circulation in dogs: effects on donor hemodynamics. AB - Controlled cross circulation (CCC) was performed in six pairs of dogs for 45 minutes with aortic cross clamping and cardioplegia. Data were collected in donor dogs at 10 minute intervals three times before, three times during, and three times after CCC and included arterial blood pressure, pulmonary capillary wedge pressure (PCWP), central venous pressure (CVP), cardiac index (CI), heart rate (HR), blood gas analysis, temperature, maximum rate of rise of left ventricular pressure dP/dt max/End diastolic volume (EDV), blood volume (BV), complete blood count (CBC) and activated clotting times (ACT). Pulse pressure (PP), systemic vascular resistance (SVR), oxygen delivery (DO2), and left ventricular cardiac work (LVCW) were calculated. Arterial blood pressure, CVP, blood gas analysis, temperature, BV, CBC, and ACT were measured in recipient dogs. During CCC, donor hemodynamic changes resembled those observed in models of acute onset arteriovenous fistulas. Insidious BV shifts can occur despite the use of occlusive roller pumps. After CCC, donor hemodynamics resembled acute blood loss, characterized by decreases in mean arterial pressure (MAP), CVP, PCWP, and CI, and increases in SVR and dP/dt max/EDV. These changes were probably caused by pump imbalance and BV shift to the recipient dog. PMID- 8719085 TI - Development of a free latissimus dorsi muscle flap in cats. AB - Anatomic and experimental evaluation of the feline latissimus dorsi muscle was performed to assess its potential use as a free muscle flap. In the anatomic study, nonselective angiography of the subscapular artery was performed in nine heparinized feline cadavers. The muscle dimensions and vascular anatomy of the dissected latissimus dorsi muscle were recorded. In the experimental study four cats underwent heterotopic transplantation of a partial latissimus dorsi flap, and three cats underwent orthotopic transplantation of a complete latissimus dorsi flap. The mean length and width of the latissimus dorsi muscle was 19.0 and 5.4 cm, respectively. The dominant vascular pedicle was the thoracodorsal artery and vein. The average length and diameter of the thoracodorsal artery was 2.7 cm and 0.6 mm, respectively. Minor vascular pedicles were provided by branches of the intercostal arteries. Numerous choke anastomoses existed between the two pedicle systems. Viability of muscle flaps based on subjective evaluation, angiography, and histopathology, was 66% and 100% in the heterotopic and orthotopic studies, respectively. Flap failure seemed to be caused by both arterial and venous thrombosis. The latissimus dorsi muscle flap met criteria required for application in microvascular reconstruction. The vascular pattern was appropriate and consistent. Donor site morbidity was low, whereas surgical accessibility was high. The muscle satisfied the physical criteria of a free flap. Long-term anastomotic patency and flap viability was shown. PMID- 8719086 TI - Evaluation of a hemostasis model for teaching basic surgical skills. AB - The need for alternative methods of teaching veterinary medicine and surgery has increased in recent years because of increasing costs and changing public opinion. For these reasons a hemostasis model was developed that mimics the arteries and veins of the peripheral vascular system, and can be used to teach the basic skills involved in blood vessel ligation and division. This study evaluated the effectiveness of the fluid hemostasis model compared with using live animals for teaching these skills. Forty sophomore veterinary students participated in the study. Two groups of 20 students each received identical instruction in the basic techniques required for vessel ligation and division. The students then completed various exercises using inanimate models to objectively evaluate their psychomotor skills. Both groups then practiced the techniques for equal time periods; one group used the hemostasis model and the other performed a splenectomy on live dogs. After the practice session, the students were videotaped (for later evaluation), as they performed vessel ligations and divisions. The students then repeated the exercises using the inanimate models for evaluation of skills improvement. Questionnaire responses before and after the project were obtained to determine the students' views on the need for inanimate models for teaching purposes. Results of this study indicate that the hemostasis model was as effective as live animals for teaching the basic skills involved in blood vessel ligation. The students' opinions regarding the use of properly designed inanimate models for teaching these skills were dramatically changed. PMID- 8719087 TI - Postoperative analgesia for stifle surgery: a comparison of intra-articular bupivacaine, morphine, or saline. AB - A prospective study was undertaken to compare the analgesic effect of intra articular bupivacaine, morphine, or saline in the 24-hour period following cranial cruciate ligament repair in dogs. Thirty-six clinical patients with ruptured cranial cruciate ligaments were randomly assigned to one of three groups. After surgical stabilization, and before skin closure, an intra-articular injection was given; group one (n = 12) received 0.5% bupivacaine HCl at 0.5 mL/kg, group two (n = 12) received morphine at 0.1 mg/kg diluted with saline to a volume of 0.5 mL/kg, and group three (n = 12) received saline at 0.5 mL/kg. Heart rate, respiratory rate, mean arterial blood pressure, cumulative pain score, visual analog pain score, and pain threshold test on both stifles were recorded preoperatively and at 0 to 6 and 24 hours postoperatively. Surgeons and pain scoring investigators were unaware of the intra-articular medication given. Supplemental analgesia, if needed, was provided in the postoperative period according to subjective assessment of patient discomfort. Postoperative pain scores were lowest in the bupivacaine group and highest in the saline group. Pain threshold, measured by applying calibrated loads to the knee, was higher postoperatively in the bupivacaine group than in the saline group. Dogs in the morphine and bupivacaine groups required less supplemental analgesia than dogs in the saline group. The local provision of analgesia reduces the need for systemic drugs with potential side effects. Both intra-articular morphine and intra articular bupivacaine provided better postoperative analgesia than intra articular saline, with intra-articular bupivacaine showing the greatest effect. PMID- 8719088 TI - Osteosarcoma eight years after total hip arthroplasty. AB - This report presents a case of osteosarcoma of the femoral diaphysis associated with chronic stimulation of the area by an unstable femoral prosthesis after total hip arthroplasty performed 8 years previously. The pathogenesis of this transformation may be similar to that proposed for fracture-associated sarcoma. Neoplasia should be considered as a possible late complication of total hip arthroplasty. PMID- 8719089 TI - Cardiopulmonary effects of using carbon dioxide for laparoscopic surgery in dogs. AB - Cardiopulmonary effects of laparoscopic surgery were investigated in five crossbred dogs (21 +/- 1.9 kg). Premedicated dogs were anesthetized with thiopental and maintained with halothane at 1.5 times minimum alveolar concentration in oxygen. Controlled ventilation maintained partial pressure of end-tidal CO2 at 40 +/- 2 mm Hg. Vecuronium was used for skeletal muscle relaxation. After instrumentation and stabilization, baseline measurements were made of cardiac output (thermodilution technique), mean systemic, mean pulmonary arterial and pulmonary wedge pressures, heart rate, saphenous vein and central venous pressures, and minute ventilation. Baseline arterial and mixed venous blood samples were drawn for analysis of pH, PaO2, PaCO2, PvO2, PvCO2, and bicarbonate concentrations. Systemic and pulmonary vascular resistances, oxygen delivery and consumption, shunt fraction, and dead space ventilation were calculated using standard formulas. Abdominal insufflation using CO2 to a pressure of 15 mm Hg for 180 minutes resulted in significant (P < .05) increases in heart rate (15 to 180 minutes), minute ventilation (75 to 135 minutes), and saphenous vein pressure (15 to 180 minutes), and decreases in pH (60 to 180 minutes) and PaO2 (60 to 180 minutes). For 30 minutes after desufflation, there was a significant decrease in PaO2, and increases in cardiac output, O2 delivery, and heart rate, compared with baseline. There was a significant increase in shunt fraction and decrease in pH at 15 minutes after desufflation only. The changes were within physiologically acceptable limits in these healthy, ventilated dogs. PMID- 8719090 TI - A comparison of epidural saline, morphine, and bupivacaine for pain relief after abdominal surgery in goats. AB - The purpose of this study was to determine the analgesic efficacy of bupivacaine, morphine, or saline (control) when injected epidurally into the lumbosacral epidural space in goats after abdominal surgery. Goats received either bupivacaine (0.5%; 1.5 mg/kg in 0.9% sodium chloride solution), 0.9% sodium chloride solution (0.2 mL/kg), or preservative-free morphine (0.1 mg/kg). Total volume injected into the epidural space was 0.2 mL/kg for all groups. The variables evaluated were times to extubation, sternal recumbency, standing, and eating; heart and respiratory rates; and pain score. Only two of the goats in the bupivacaine group were able to stand on their hindlimbs before 6 hours. Time to eating was shorter for the saline group when compared with the bupivacaine group. Heart rate over all time in the saline group (137 +/- 4 beats/min, mean +/- SEM) was higher than the morphine (125 +/- 3 beats/min) and bupivacaine groups (121 +/ 3 beats/min). Respiratory rate over all time was increased in the saline group (26 +/- 1 breaths/min) compared with the bupivacaine (24 +/- 1 breaths/min) or morphine (24 +/- 1 breaths/min) groups. At 50 minutes, the pain score for the saline group was higher than the morphine group. Pain score over all time in the saline group (1.5 +/- 0.10) was higher than the morphine (1.2 +/- 0.07) and bupivacaine (1.2 +/- 0.04) groups. One goat in the saline group required two intravenous injections of flunixin meglumine for pain. PMID- 8719091 TI - Metabolic, hormonal, and hemodynamic changes during dopamine infusions in halothane anesthetized horses. AB - Selected metabolites, hormones and cardiovascular variables were measured in halothane anesthetized horses during 1 hour of dopamine infusion at a rate of 5 micrograms/kg/min (low) and 10 micrograms/kg/min (high), and for 1 hour after infusion. Plasma cortisol increased twofold in the low-infusion group but did not change significantly in the high-infusion group. Plasma nonesterified fatty acids, blood glucose, blood lactate, and plasma insulin increased in the high infusion group. There was little difference in heart rate, systolic, diastolic, and mean arterial blood pressure between the two groups. The high infusion was associated with arrhythmias in several horses, and one horse showed ventricular fibrillation and died. If metabolic and hormonal changes are used as markers of a "stress response" in anesthetized horses the results must be carefully interpreted if a sympathomimetic agent such as dopamine is administered to maintain cardiovascular stability. PMID- 8719092 TI - Poverty and race often determine lung cancer survival, study shows. PMID- 8719093 TI - New Jersey researchers testing unique blood substitute. PMID- 8719094 TI - MRI may reduce the number of biopsies for breast cancer. PMID- 8719095 TI - Management of malignant biliary obstruction: nonoperative and palliative techniques. AB - Biliary tract drainage, with or without placement of an endoprosthesis, is used as a palliative therapy for malignant biliary obstruction. The first truly internal endoprostheses represented a distinct improvement over internal-external catheters but still remained patent for only 4 to 6 months. Metallic stents have a long-term patency of 6 to 8 months. At present, it appears that patients with unresectable pancreatic cancer should be palliated with endoscopically placed plastic or metal stents, whereas those with malignant obstructions higher in the biliary tree are probably better managed with transhepatically placed stents. The combination of brachytherapy plus external-beam radiation followed by implantation of a Gianturco metal stent may be a viable approach to treating obstructions in patients with cholangiocarcinoma. For those with other noncholangiocarcinomas, particularly when life expectancy exceeds anticipated stent patency duration, the Wallstent may be the device of choice. PMID- 8719096 TI - Clinical trials at the medicine branch of the NCI-part 2. PMID- 8719097 TI - Invasive radiologic procedures pose low risk of HIV transmission from doctor to patient. PMID- 8719098 TI - Vinorelbine/paclitaxel combination studied in treatment of metastatic breast cancer patients. PMID- 8719099 TI - Measuring the cost-effectiveness of cancer care. AB - Historically, new therapeutic strategies for cancer have been evaluated on the basis of safety and clinical efficacy. However, the current national emphasis on efficiency of resource allocation has led to the inclusion of economic assessments in oncology studies. Economic assessments measure patients' health status and resource consumption associated with a therapeutic strategy, and combine these in a cost-effectiveness analysis. Study design can include prospective analysis of clinical trials, retrospective analysis of a clinical trial or administrative databases, or a decision analytic model. Economic analysis is being used increasingly in oncology and will continue to provide meaningful data to assist clinicians in determining the optimal treatment strategies for cancer patients and to help inform health policy decision-makers about the importance of specific cancer therapeutic strategies. PMID- 8719100 TI - Current concepts in surgical management of neck metastases from head and neck cancer. AB - The surgical management of neck metastases from head and neck cancer consists primarily of neck dissection. An awareness of both the cervical anatomy and natural history of squamous cell carcinoma of the upper aerodigestive tract is necessary to understand the role of neck dissection and to appreciate when it may be appropriate to modify the standard radical neck dissection. The use of imaging to augment palpation of the neck has resulted in greater accuracy in identifying neck metastases and has further clarified the role of modified neck dissections. Depending on the site and size of the primary tumor, radiation therapy, modified neck dissection, or selective neck dissection are all appropriate options for the elective treatment of the neck when there is a high risk of occult metastasis. Therapeutic neck dissection alone is adequate for early-stage neck disease, but must be combined with irradiation for more advanced stages. Regional control of tumor metastases is highly dependent on the stage of neck disease and the presence of extranodal cancer spread. PMID- 8719101 TI - PSA-based diagnoses are leading to improved prostate cancer outcomes. PMID- 8719102 TI - Integration of vinorelbine into chemotherapy strategies for non-small-cell lung cancer. AB - Meta-analyses of randomized clinical studies comparing combination chemotherapy versus "best supportive care" for advanced non-small-cell lung cancer have revealed a small, but statistically significant survival advantage for patients who receive chemotherapy. However, overall increases in lifespan have been short, and the great majority of patients die within 1 year of diagnosis. In the last few years, several new drugs with promising activity have been identified. Of these, vinorelbine has already been shown to increase survival rates in randomized clinical trials. In particular, one such trial showed the combination of vinorelbine and cisplatin to result in statistically superior survival rates, compared with "standard" therapy of cisplatin and vindesine, and with single agent vinorelbine. A second study comparing vinorelbine to fluorouracil/leucovorin also demonstrated a survival benefit for patients treated with vinorelbine. Therefore, the combination of vinorelbine and cisplatin represents one new option for initial therapy of newly diagnosed stage IV non small-cell lung cancer. PMID- 8719103 TI - Inhibition of smooth muscle cell proliferation in vitro by leflunomide, a new immunosuppressant, is antagonized by uridine. AB - Chronic rejection in the form of graft vascular disease (GVD) continues to plague clinical transplantation of vascularized organs. The histopathology of this lesion is characterized by neointimal hyperplasia, smooth muscle cell proliferation, and obliterative arteriopathy. Due to the lack of effective medical therapy for preventing or reversing these chronic vascular changes, retransplantation remains the final resort in treatment. Some of the newer immunosuppressive agents, including the new isoxazole derivative leflunomide (LFM), have shown efficacy in preventing chronic rejection in animal models of transplantation. Although its mechanism of action remains incompletely elucidated, previous work using lymphocytes in vitro suggests that the drug might act as a tyrosine kinase inhibitor, an inhibitor of de novo pyrimidine biosynthesis, or both. In order to elucidate whether the efficacy of LFM in vivo is attributable not only to anti-proliferative effects on the recipient immune system but also to direct effects on mesenchymal cells in the donor organ, we examined the effects of LFM on a transformed 9E11G murine smooth muscle cell (M SMC) line in vitro. We demonstrate here that the active metabolite of LFM, A77 1726, dose-dependently inhibits the constitutive and growth-factor stimulated proliferation of M-SMC in vitro. Furthermore, the anti-proliferative effect of the drug can be reversed by the addition of uridine to the culture medium. These results suggest that inhibition of uridine biosynthesis appears to be a mechanism by which LFM exerts anti-proliferative effects on both lymphocytes and smooth muscle cells, and this dual action may be responsible for its efficacy in preventing GVD in vivo. PMID- 8719104 TI - Long-lasting functional unresponsiveness induced by a milk-transmitted Mls-1a like superantigen. AB - We have recently shown (Piazzon et al. (1994) J. Immunol. 153, 1553) that foster nursing of BALB/c mice on F1 Mls-1bxa mothers induce the progressive deletion of V beta 6+ and 8.1+ T cells in 50% of the mice. Preceding clonal deletion, a state of functional inactivation of CD4+ T cells to Mls-1a and anti-V beta 6 antibodies was detected in young mice. In the present paper we show that foster-nursing of BALB/c mice on (BALB/cxAKR)FI mothers is able to induce alterations in T cell reactivity in the non-deletor mice. Lymph node cells from foster-nursed mice show a decreased proliferative level against anti-V beta 6 antibodies and a diminished response in MLR and in CTL assays. The proliferative responses to either OVA or Con-A are also reduced. This state of functional inactivation is detected even in 6-month-old foster-nursed mice. Thus, the transmission through milk of the Mls-1a like superantigen correlates in the non-deletor mice with a long-lasting state of functional inactivation and a decreased immune reactivity. PMID- 8719105 TI - Human lymphocytes are more susceptible to measles virus than granulocytes, which is attributable to the phenotypic differences of their membrane cofactor protein (CD46). AB - Membrane cofactor protein (MCP, CD46) of the complement system is a measles virus (MV) receptor. Human lymphocytes express a heavily glycosylated (H) and a lightly glycosylated (L) form of MCP, which confers a two-band profile on SDS-PAGE the ratio of which is controlled genetically and organ-specifically. In contrast, granulocytes express a single heavily glycosylated form regardless of lymphocyte MCP phenotype. We investigated susceptibility to MV of granulocytes and lymphocytes from individuals with different lymphocyte MCP phenotypes. In any individual, granulocytes were > 10-fold less susceptible to MV than lymphocytes, and the lymphocytes with predominant H form were generally less susceptible to those with an increasing amount of L form. Thus, lymphocytes always exhibit high susceptibility to MV compared to granulocytes in all individuals. This finding may explain the lymphopenia and immunosuppression observed secondary to MV infection. PMID- 8719106 TI - Anti-gamma delta T cell antibody blocks the induction and maintenance of oral tolerance to ovalbumin in mice. AB - The oral administration of antigens is one of the means of inducing tolerance in adult mammals. In this report, the role of gamma delta T cells in the induction and maintenance of orally-induced tolerance to ovalbumin was investigated. The injection of a monoclonal anti-gamma delta T cell monoclonal antibody blocked the induction of oral tolerance, because the secondary immune responses to ovalbumin in these animals were comparable to the corresponding responses in ovalbumin immunized control mice. Furthermore, depletion of gamma delta T cells either in vivo or in vitro abolished already established oral-tolerance. The fact that the state of tolerance could be adoptively transferred to naive recipients by CD3+ alpha beta- gamma delta + spleen cells from tolerant mice. These results suggest that systemic oral tolerance is induced and actively maintained by mechanisms involving gamma delta T cells. PMID- 8719107 TI - Glutathione S-transferase a major allergen of the house dust mite, Dermatophagoides pteronyssinus. AB - Recently, we cloned a new Dermatophagoides pteronyssinus allergen homologous with glutathione S-transferase. IgE radioimmunoassays against the Escherichia coli lysate of the recombinant clone reveal that 40% of mite allergic subjects recognize the mite glutathione S-transferase allergen. Of these sera, greater that 50% have reactivity with the recombinant mite glutathione S-transferase that is greater than 10 times the result observed with a normal control. Immunoblotting studies with sera from patients that recognize the recombinant protein reveal IgE binding to a 26-kDa protein on immunoblots of reduced mite protein extracts. The 26-kDa IgE-binding band observed on immunoblots of reduced mite proteins, corresponding to the cloned protein, is a separate and unique allergen from the 25-kDa Der p I as the apparent electrophoretic molecular mass of Der p I shifts from 25 to 30 kDa under conditions of reduction. PMID- 8719108 TI - The bone marrow as a site of antibody production after a mucosal immunization. AB - To study the importance of the bone marrow in the production of specific antibodies after a mucosal immunization with cholera toxin, the IgG, IgA and IgM specific antibody forming cells were evaluated by ELISPOT in Peyer patches, mesenteric lymph node (MLN), spleen, blood and bone marrow (BM). When 50-day-old rats were immunized intra-Peyer patches, a similar number of IgG and IgA antitoxin antibody forming cells (AFC) were found in the BM, whereas in the other lymphoid tissues a higher number of IgG antitoxin AFC were found. In all sites the peak of AFC was obtained 2 weeks after immunization. The administration of CT to 35-week-old rats resulted in a stronger immune response in all lymphoid tissues studied, but the proportion of antitoxin AFC contributed by the BM had not changed. One oral dose of cholera toxin resulted in a low number of antitoxin AFC, whereas when two or three doses of CT were administered orally an increase in the number of AFC was observed in the BM, reaching similar or higher numbers of IgG and IgA AFC than in the spleen. In all cases the highest number of AFC/10(6) cells was observed in the MLN, whereas antitoxin AFC were not found in the blood. The total number of AFC recovered from each organ was calculated taken into account that the BM of one femur represents 9% of the total BM. So, it was found that the BM is an important site in the production of IgG antitoxin antibodies, being the main site in the IgA antitoxin antibody production. PMID- 8719109 TI - Prevention of type I diabetes in the non-obese diabetic (NOD) mouse with 15 deoxyspergualin (15-DS) or 15-DS + cyclosporin A (CyA). AB - By selectively inbreeding diabetic individuals, we have been able to establish an NOD mouse population with a genetic predisposition towards insulin-dependent diabetes mellitus (IDDM) in approximately 100% of cases. We examined the preventive effect of 15-DS or 15DS + CyA on developing IDDM in these animals. Whereas 15-DS has been proved to be effective in preventing diabetes (significant decrease of the diabetic risk ratio to 0.368 and a reduction of the incidence of the disease to 46.7%), combined treatment with CyA did not produce any additional benefit. PMID- 8719110 TI - Monocyte derived IL 10 and PGE2 are associated with the absence of Th 1 cells and in vitro T cell suppression in lepromatous leprosy. AB - Our previous studies had shown that the clinicopathological spectrum in leprosy was associated with discrete T cell subsets in circulation, with tuberculoid patients having antigen-induced Th 1, whereas lepromatous leprosy patients with antigen-specific T cell anergy possessed Th 2 cells. The present study shows that infected monocytes from lepromatous but not tuberculoid leprosy patients released soluble factors (MoF(s)) containing IL-10 and PGE2 which inhibited M. leprae induced in vitro lymphoproliferation of previously sensitised healthy or tuberculoid leprosy subjects. A strong negative correlation was observed between adherent cell derived IL-10 and IL-2 at the level of both the product and cytokine mRNA. Moreover, anti-IL-10 antibodies and indomethacin partially reversed the suppressor effects of MoF(s). Taken together these studies indicate that infected monocytes contribute to the development of T cell anergy by releasing factors that affect regulatory cytokines and T cell subset differentiation in lepromatous leprosy. PMID- 8719111 TI - Anti-actin antibodies in sera from patients with autoimmune liver diseases and patients with carcinomas by ELISA. AB - Smooth muscle antibodies (SMA) were initially detected in sera of patients with chronic active hepatitis (CAH). Subsequently, their presence was demonstrated in a wide variety of other diseases. SMA are a mixture of antibodies directed towards different cytoskeletal antigens. Sera with high titers of anti-actin antibodies (AAA), a subgroup of SMA, are most frequently found among patients with autoimmune chronic active hepatitis (CAH) and, to a lesser extent, among patients with primary biliary cirrhosis (PBC) and other diseases. It is therefore established that AAA are a reliable marker to autoimmune CAH. The purpose of this study was to determine the titer of AAA in sera of patients with liver, autoimmune diseases and carcinomas, using the enzyme-linked immunoabsorbent assay (ELISA) method. The results were expressed as the optical density (OD) of the examined sera divided by the OD of a sera from a healthy control (presented as percentages +/- standard deviation). Sera of 33 patients with cirrhosis, nine patients with autoimmune CAH, fifteen patients with non-autoimmune CAH, eight patients with PBC, 30 patients with Sjogren's syndrome, 60 patients with SLE, 142 patients with carcinomas of different kinds, and 34 healthy donors were examined for the presence of AAA by ELISA. Statistically significant (P < 0.003) higher titers of AAA were detected in patients with autoimmune CAH (57 +/- 23%) compared with the control group and to other groups of diseases. AAA titers in non autoimmune CAH were not significantly higher compared to the control group. High titers of AAA were detected in 67% of the patients with autoimmune CAH, as compared with other diseases in which only up to 13% of the patients exhibited AAA positivity. CONCLUSION: existence of higher levels of AAA noted in 67% of the patients with autoimmune CAH with regard to other groups of diseases, emphasizes the value of AAA as sensitive and specific markers, capable of characterizing the patients with autoimmune chronic active hepatitis. PMID- 8719112 TI - Effect of pre-S1 antigen on human lymphocyte proliferative responses. AB - The peripheral blood mononuclear cells (PBMCs) from patients acutely infected with HBV and recovered completely (n = 20), patients with chronic hepatitis B infection (CHB)- (n = 10) and HBsAg-positive carriers (n = 9) and healthy individuals (n = 8) were studied for their in vitro proliferative response to a synthetic pre-S1(20-49)x4 antigen. PBMCs from convalescents showed significant proliferative response in the presence of synthetic pre-S1 antigen. PBMCs from CHB- and HBsAg-positive exhibited reduced proliferative response not only to Pre S1 antigen but also to nonspecific mitogens. This study suggests that the immune recognition of pre-S1 antigen and response of PBMCs to the pre-S1 antigen may be an important part of the normal human response to HBV infection. Failure to clear the HBV infection with development of the chronic carrier state may be caused by the lack of an efficient pre-S1 antigen-specific response. PMID- 8719113 TI - Analysis of mRNA expression for interleukin-1 genes on human testicular cells. AB - We have investigated mRNA expression for IL-1 alpha and IL-1 beta gene on fractionated human testicular cells. Using RT-PCR and Northern blot hybridization technique we detected the presence of IL-1 alpha transcripts, predominantly in the intratubular compartment of the testis, comprising gametogenic and Sertoli cells. We were also able to detect mRNA for IL-1 alpha on the testicular interstitium, but at significantly lower levels. The intertubular compartment of the testis, mainly consisting of macrophages and Leydig cells, appeared however, to be a site for IL-1 beta gene expression. Our experimental data confirm previous results obtained in animal models indicating that the testis is capable of producing interleukin-1 under physiological conditions. Testicular IL-1 may function as a tissue-specific factor modulating both spermato- and steroidogenic activity of human testis. PMID- 8719114 TI - Interleukin-1 beta induces the expression of hsp70, heme oxygenase and Mn-SOD in FACS-purified rat islet beta-cells, but not in alpha-cells. AB - The cytokine IL-1 beta has previously been demonstrated to induce the expression of the stress genes iNOS, hsp70, heme oxygenase and Mn-SOD in rat pancreatic islets in vitro. The aim of this study was to determine whether the IL-1 beta induced effects are specific for the insulin producing beta-cell, or whether other islet cells, such as the glucagon-producing alpha-cell, respond to IL-1 beta addition. Purified rat alpha- and beta-cell suspensions were obtained by fluorescence-activated cell sorting and incubated with or without IL-1 beta (25 U/ml) for 24 h. The alpha- and beta-cell contents of hsp70, heme oxygenase and Mn SOD and medium nitrite levels were determined. It was found that IL-1 beta exposure induced the production of nitric oxide in beta-cells, but not in alpha cells. Moreover, the expression of hsp70, heme oxygenase and Mn-SOD was also induced in beta-cells, but not in alpha-cells. There were no detectable levels of hsp70 in alpha-cells. It is concluded that the stress gene response following IL 1 beta exposure is markedly different in alpha- and beta-cells. This finding may be of importance for the understanding of the autoimmune destruction of beta cells in insulin-dependent diabetes mellitus. PMID- 8719115 TI - Systemic in vivo antitumor activity of interleukin-12 against both transplantable and primary tumor. PMID- 8719116 TI - Impact of host genetic background on bacillus Calmette-Guerin infection in naturally susceptible mice. PMID- 8719117 TI - Persistent non-B cell lymphocytosis in HIV-infected individuals. PMID- 8719118 TI - Neuroleptic malignant syndrome and catatonia: one entity or two? PMID- 8719119 TI - The genes encoding the 5HT1D alpha and 5HT1D beta receptors are unchanged in patients with panic disorder. AB - To determine the serotonergic function in panic disorder, sequencing of the genes encoding the 5HT1D alpha and 5HT1D beta receptors was carried out. The genes for the 5HT1D alpha and 5HT1D beta receptors were amplified by polymerase chain reaction and sequenced by the dideoxy method. Some patients have both nucleotides C and T at position 1080 in 5HT1D alpha receptor gene; however, both of them code the same amino acid, asparagine. The 5HT1D beta receptor gene had a substitution from GCA276 to GCG276, not only panic disorder but also in controls; however, this substitution does not change the corresponding amino acid, alanine92. PMID- 8719120 TI - Hypertonic hemolysis in patients with a mood disorder. AB - Hypertonic hemolysis was increased in patients with a manic episode (n = 6, mean NaCl concentration in mol/L at which 50% of the erythrocytes was hemolyzed (H50) 3.24 +/- 0.10), compared to healthy controls (n = 12, mean H50 3.43 +/- 0.13) (p < or = 0.02) and patients with a nonaffective psychotic disorder (n = 15, mean H50 3.42 +/- 0.18) (p < or = 0.02). Compared to these two control groups, hypertonic hemolysis was decreased in patients with a severe depressive episode (n = 8, mean H50 3.62 +/- 0.19) (p < or = 0.05 and p < or = 0.02, respectively). Within the total depressed patient group, the patients with an increased genetic vulnerability (n = 8, mean H50 3.68 +/- 0.16) showed a decreased hypertonic hemolysis compared to the other depressed patients (n = 14, mean H50 3.40 +/- 0.16) (p < or = 0.002). PMID- 8719121 TI - Bright light therapy stabilizes the antidepressant effect of partial sleep deprivation. AB - Partial sleep deprivation (PSD) results in a pronounced decrease of depressive symptoms in the majority of patients with major depressive disorder. Generally this acute antidepressant effect is not stable, relapse usually occurs after one night of recovery sleep. We therefore studied whether light therapy, beginning in the morning after PSD, is able to prevent the relapse after sleep deprivation, using a controlled, balanced, parallel design. All patients received an antidepressant medication, which was kept constant before and during the study period. Fourteen of 20 patients (70%) showed a reduction of at least 40% in the Hamilton Depression Rating Scale (HDRS) in the morning after PSD and were classified as PSD responders. Responders as well as nonresponders were randomly assigned to receive either bright light (BL/3000 lux) or dim light (DL/100 lux) therapy during the following 6 days after PSD. In the responder group BL therapy prevented significantly (p = 0.005) the relapse after the next night of sleep and prolonged significantly (p = 0.011) the antidepressant effects of PSD up to 7 days. In contrast, patients in the DL condition relapsed after the recovery night and showed no further improvement of the depressive syndrome after 1 week of DL therapy. PSD nonresponders did not benefit from light treatment. These findings indicate that BL therapy might be efficacious to prevent relapse after PSD. PMID- 8719122 TI - Light attenuation by the human eyelid. AB - Although it has been reported that light treatment during sleep can modify biological rhythms, the amount of light that is transmitted through the human eyelid has not been established. We evaluated eyelid transmission with a visual threshold response. Estimated light transmission through the eyelids was 0.3% for blue, 0.3% for green, and 5.6% for red light. The eyelid was an effective attenuator and acted as a red-pass filter. Illumination intensity and color balance after passing through the eyelid should be considered in evaluating the effects of light treatments during sleep. PMID- 8719123 TI - Subjective responses to d-amphetamine alone and after pimozide pretreatment in normal, healthy volunteers. AB - The role of dopamine in d-amphetamine-induced euphoria has not been systematically examined in normal volunteers. Therefore, we examined the effects of the dopamine antagonist, pimozide, on responses to d-amphetamine in healthy volunteers, using a within-subjects, double-blind design. Ten subjects received single oral doses of d-amphetamine (0, 10, 20 mg) 2 hours following pretreatment with pimozide (0, 1, 2 mg). Subjective, behavioral, and physiological effects were assessed predrug and for 3 hours after d-amphetamine administration. d Amphetamine alone produced prototypic effects on a variety of measures, including euphoria and drug liking. Pimozide did not produce any effects when administered alone and produced inconsistent effects on responses to d-amphetamine. Although higher doses of pimozide may be needed to antagonize the euphorigenic effects of d-amphetamine, these results raise the possibility that the role of dopamine in the subjective effects of stimulants may be more complex than initially appreciated. PMID- 8719124 TI - Sensorimotor gating in boys with Tourette's syndrome and ADHD: preliminary results. AB - Deficits in sensorimotor gating, defined by prepulse inhibition (PPI), have been associated with subcortical dopaminergic overactivity in animal and clinical studies. Utilizing supraorbital nerve electrical stimulation, we produced adequate blink responses and measured decreases in amplitude resulting from electric prestimuli just above sensory threshold. Seven boys comorbid for attention-deficit hyperactivity disorder (ADHD) and a tic disorder had significantly reduced PPI, compared to 14 screened controls and seven boys with ADHD alone. If independently replicated, these results may reflect greater neurologic immaturity in these comorbid subjects. Alternatively, these findings, together with other converging lines of evidence, suggest that deficient pallidal inhibition may be etiologically related to tic and movement disorders. PMID- 8719125 TI - Sleep and dreams in Vietnam PTSD and depression. AB - The present study compares the sleep and dreams of three groups of subjects: 1) Vietnam veterans with posttraumatic stress disorder (PTSD) and major depression, 2) veterans with depression alone, and 3) veterans with neither PTSD nor depression (i.e., normal controls). Sleep recordings indicate only one significant difference between the PTSD/depressed and depressed alone groups: sleep latency was prolonged in the depressed alone patients compared with the other two groups. The two patient groups differed from controls in the manner already reported for depressed patients (decreased REM latency, increased REM density, reduced total sleep time, reduced sleep efficiency), with some of the differences significant only at the trend level. Dreams were obtained from REM awakenings. Dream recall rate and report length did not differ between groups. Mean anxiety level in dreams was less than 1 (mild) for all three groups, with major depression patients scoring significantly higher than controls. Dreams of PTSD/depressed patients were significantly less likely to be set in the present than dreams of the other two groups. PMID- 8719126 TI - Gender differences in gating of the auditory evoked potential in normal subjects. AB - Central nervous system (CNS) inhibitory mechanisms hypothesized to "gate" repetitive sensory inputs have been implicated in the pathology of schizophrenia. The present study investigated gender differences in inhibitory gating of evoked brain responses to repeated stimuli in normal subjects (30 women and 30 men) using an auditory conditioning-testing paradigm. Pairs of click stimuli (S1 and S2) were presented with a 0.5 s intrapair and a 10 s interpair interval. The amplitudes and latencies of the P50, N100, P180 components of the auditory evoked response to the conditioning (S1) and test response (S2) were measured, and the gating ratios were computed (T/C ratio = S2/S1 * 100). The amplitudes to S1 were not significantly different between men and women at P50, N100, or P180. However, women had significantly higher amplitudes to S2 at P50 (p = 0.03) and N100 (p = 0.04). The T/C ratios for women were higher (i.e., less suppression of response to S2) for P50 (p = 0.08) and N100 (p = 0.04) compared to men. The results suggested that differences in auditory gating between men and women were not due to biological differences in the P50 and N100 generators but possibly to differential influence of inhibitory mechanisms acting on the generator substrates of these evoked responses. PMID- 8719128 TI - Evaluating cumulated research. I: The inadequacy of traditional methods. AB - Virtually all reviews of cumulated studies rely on statistical significance as a criterion for evaluating the reproducibility of the phenomenon under review. Despite its nearly universal application, that criterion is entirely inadequate: Its application is very likely to lead a reviewer to conclude that a phenomenon does not discriminate patients from controls when, in fact, it does do so. The reviewer is, paradoxically, more likely to draw this incorrect conclusion as more studies become available for review. It can lead a reviewer to conclude that one phenomenon is more discriminating than another when the opposite is actually true. Fortunately, procedures that do not distort the review process are available; some of these are briefly discussed. PMID- 8719127 TI - Affective reactivity of cognitive functioning and family history in schizophrenia. AB - This article reports the results of two related studies investigating the association between affective reactivity of cognitive functioning in schizophrenia and family psychiatric history. In Study #1, we examined affective reactivity of positive formal thought disorder symptoms in 29 schizophrenic inpatients. We found that thought disorder was greatly exacerbated by negative affect in those patients with a family history of schizophrenia (SFH) (n = 11), and not in those without the family history (SNFH) (n = 18). In Study #2, we replicated this finding with a stable outpatient sample (n = 10). We also administered dichotic listening tests using affectively neutral and affectively negative stimuli, and found that right-ear advantage was more markedly diminished on the affectively negative task than on the neutral task in the SFH (n = 6) but not the SNFH (n = 4) subjects. These findings support our hypothesis that cognitive symptoms in schizophrenia are exacerbated by negative affect, and that this affective reactivity of symptoms is associated with a familial form of the disorder. PMID- 8719129 TI - Dietary purine nucleotides and the gastrointestinal tract. AB - There is a growing body of research that demonstrates a role for dietary nucleotides, the building blocks of RNA and DNA, during intestinal development, turnover, and repair. There is evidence that the effects of purine nucleotides may be mediated through intestinal gene transcription and, more specifically, through the action of transcription factors that, at least sometimes, are directly related to nutrition. Some of the suggested specific roles for dietary nucleotides include the enhancement of the normal host defense system, effects on neonatal lipid metabolism, and influence on iron bioavailability. This review focuses on the potential role of dietary (purine) nucleotides in the maintenance of intestinal integrity and reviews potential mechanisms for these effects. PMID- 8719130 TI - How valid are our reference standards of nutrition? AB - The validity of anthropometric measurements in the assessment of nutritional status depends on the use of appropriate standards. The most commonly used standards for triceps skinfold thickness and mid-arm muscle circumferences are based on Jelliffe's and Frisancho's tables. In this study we compared these two standards in a population of healthy subjects and patients with a variety of pathological disorders. The study showed that the correlation between these two standards was poor. When Frisancho's standards were used as "gold standards," the positive predictive value of Jelliffe's standards for triceps skinfold thickness was only 22% and for mid-arm muscle circumference only 53%; the false positive results for triceps skinfold thickness and mid-arm muscle circumference were 28% and 27%, respectively. This study emphasizes the need to develop appropriate standards for the studied population. Until such standards are available, workers assessing nutritional status in population studies would be advised to interpret their findings with caution, and, on the basis of this study, we recommend the use of Frisancho's standard in preference to Jelliffe's standards. PMID- 8719131 TI - Fat emulsions as an ideal nonprotein energy source under surgical stress for diabetic patients. AB - We studied the possibility of fat emulsions being a major calorie source, and if possible, a new infusion protocol without glucose for diabetic patients undergoing surgery. High dosages of fat emulsions and amino acids (HFHA infusions) were investigated as a substitute for conventional glucose-based infusions for 7 days after surgery. Without insulin and glucose, blood glucose was kept within the normal range, in spite of the fact that patients were recovering from major surgeries. Blood triglyceride and fatty free acids (FFA) increased after the start of the HFHA infusions but remained within normal range. Liver function tests and nutritional states estimated by nitrogen balance or rapid turnover proteins were no different from those of the conventional glucose based infusion, but the significance of the HFHA infusion lies in the fact that it was applied to patients in whom it was impossible to administer enough calories by using the conventional glucose-based infusion, even if insulin was used. This HFHA infusion, which enables easy blood glucose control, sufficient calorie supply without complications, and is also nutritionally equivalent to the conventional glucose-based infusion, can be a new nutritional option for postoperative patients with impaired glucose tolerance. PMID- 8719132 TI - Energy and nitrogen balance and changes in midupper-arm circumference with multiple organ failure. AB - Energy intake and energy expenditure, nitrogen intake, and urinary nitrogen excretion (or urea production rates) were measured in 35 intravenously fed patients with multiple organ failure over the course of their illness to determine to what extent nutrient requirements were met despite fluid retention. Energy and nitrogen balance were related to serial measurements of midupper-arm circumference (MAC). The target feeding regimen of 176 kJ (42 kcal)/kg fat-free mass (FFM) was achieved in only three patients and the target of 0.24 g N/kg FFM in only four. Two patterns of change in MAC were noted: a steady decrease with time and no change with time. Serial muscle biopsy data indicated that all the patients were wasting away; the maintenance of MAC in the group with no change over time was due to fluid retention. Abnormal losses were not measured, but energy and nitrogen balance in the group in which arm circumference decreased had no apparent effect on the rate of wasting. PMID- 8719133 TI - Controlling bacterial contamination of an enteral formula through the use of a unique closed system: contamination, enteral formulas, closed system. AB - Contamination of enteral diets may play an essential role in formula tolerance and safety for patients. Contaminated enteral formula commonly support microbiological growth. Commercially sterile liquid formulas received from the manufacturer are required by the Food & Drug Administration (FDA) to be shelf stable and free from enteric pathogens. This study examined the use of large volume, closed system containers in a typical nursing home. Large volume (1500 mL) containers with unique pierceable caps and piercing spikes were studied to determine their ability to reduce the incidence of microbiological contamination due to their design and ability to decrease handling requirements. This study took place in a room of a typical nursing home. In this clinical setting, 211 containers and administration spike sets were evaluated following a 36-h hangtime. Contamination was virtually nondetectable. Nursing staff in a clinical facility can effectively utilize a large volume, prefilled, ready-to-use feeding system to achieve delivery of noncontaminated product for up to 36 h hangtime. PMID- 8719134 TI - Inhibition of small intestinal motility by casein: a role of beta casomorphins? AB - Several aspects on the effect of nutrients on small intestinal motility are not completely understood. We have analyzed changes of motor activity of the canine small bowel following intragastric administration of casein and soy protein. Studies were performed by intraluminal manometry with external transducers and multiple infused catheters with closely spaced registrating sites. Casein was followed by a statistically significant decrease of amplitude and frequency of small intestinal contractions, compared to soy protein. Pretreatment with naloxone suppressed the inhibitory effect of casein, suggesting that stimulation of opioid receptors by beta casomorphins, a product of digestion of casein, might be involved in the motility changes observed. PMID- 8719135 TI - Diets high in polyunsaturated oils decrease lymphocyte membrane anisotropy and B cell mitogenesis. AB - Balb/c mice were fed semipurified diets containing 40% of total calories as lipids: olive, safflower, or fish oil. After 2 weeks of feeding, splenic mononuclear cells were isolated and assayed for steady-state fluorescence anisotropy by means of the probe diphenylhexatriene. Splenocytes were also incubated with the mitogens concanavalin A, phytohemagglutinin, or lipopolysaccharide; subsequent uptake of tritiated thymidine was measured as an index of proliferation. No differences were observed in the membrane anisotropy and mitogenesis of mice fed the olive oil diet (high in monounsaturated fatty acids) and the control group. However, membrane anisotropy was significantly lowered in mice fed the safflower or fish oil diets (high in polyunsaturated fatty acids), and the mitogenic response to lipopolysaccharide was suppressed. Mitogenic responses to concanavalin A and phytohemagglutinin were not significantly affected by the type of lipid in the diet. PMID- 8719136 TI - An infant with intractable Crohn's disease: a case report. PMID- 8719137 TI - Diabetic renal disease: microalbuminuria, implications and intervention. AB - Diabetic nephropathy is the leading cause of end-stage renal failure in the developed world. Proteinuria ("macroalbuminuria" > 200 micrograms/min; or 300 mg/24 h) heralds a phase of established renal pathology with inexorable decline to end-stage renal disease, although its progression can be delayed by antihypertensive medication, in particular the angiotensin-converting enzyme inhibitors (ACEI). Control of blood pressure is vital; even if in the normal range, it is usually raised compared with that in nondiabetic control groups. Reducing blood pressure can lower the rate of decline of the glomerular filtration rate by 90%. Before established proteinuria there is a "microalbuminuric" (20-20 micrograms/min, or 30-300 mg/24 h) phase, and during this time preventive intervention may be effective. In so-called "normotensive" microalbuminuric subjects antihypertensive medications, in particular the ACEI, significantly reduce the progression to macroalbuminuria. Control of glycemia is important; recent evidence has shown that it is particularly important before the development of microalbuminuria; thereafter the role of glycemic control is not clear. Some researchers have suggested that protein restriction may be helpful, but more data are required. For the moment, improved glycemic control in the normoalbuminuric diabetic subjects and treatment with ACEI after the onset of microalbuminuria would seem appropriate in light of knowledge today. Furthermore, any level of hypertension is totally unacceptable and should be treated aggressively; the ACEI seem to be becoming the "agents of choice." PMID- 8719138 TI - Critical pathways for improved outcome using nutrition support. PMID- 8719139 TI - Pharmaceutical aspects of parenteral nutrition: a historical perspective. PMID- 8719140 TI - Nutrition and body composition in multiple organ failure. PMID- 8719141 TI - Medicaid costs drive state health care reform--Part II. PMID- 8719142 TI - Robustness and power. PMID- 8719143 TI - Muscle proteolysis induced by a circulating peptide in patients with sepsis or trauma. 1983. PMID- 8719144 TI - Breast screening in Britain. PMID- 8719145 TI - Monitoring interval cancers in breast screening programmes: the east Anglian experience. Quality Assurance Management Group of the East Anglian Breast Screening Programme. AB - Interval cancer rates are a major determinant of the success of a screening programme. In the Swedish two county study, on which the United Kingdom programme is based, a 39% reduction in mortality was observed in screened women aged 50-64. Using data from the Swedish study, the relationship between interval cancer incidence and the likely future effect on breast cancer mortality was quantified. In East Anglia, as elsewhere in the United Kingdom, interval cancers rates are nearly double those obtained in Sweden: interval cancer rates in the first, second, and third years respectively, after a negative screen were 24%, 59%, and 79% of the expected underlying incidence in the absence of screening. The corresponding figures from the two county study were 17%, 30%, and 56%. From these it was estimated that the mortality reduction in East Anglia will be 21%, which is lower than the 35% observed in invited women in this age group in the Swedish two county study and the 25% specified in the Health of the Nation target. In a rereading exercise, using screening mammograms from women who were screen normal, who had screen detected cancers, or who subsequently developed interval cancers, four out of five radiologists recommended recall for around 70% of the original mammograms (classed as screen normal at time of screening) from 33 interval cancers. This suggests that sensitivity is a contributory factor to the higher interval cancer rates in East Anglia. PMID- 8719146 TI - Results from the NHS breast screening programme 1990-1993. AB - OBJECTIVE: To present results from the NHS breast screening programme (NHSBSP) for the three year period 1990 to 1993, and to examine the extent to which interim targets are being met. METHODS: Data have been collated from all screening programmes in the United Kingdom on standard "Korner" returns, supplemented for the year 1991/92 by data from the radiology quality assurance programme. Most of the data refer to the prevalent screening round, but some data on rescreening are also available. RESULTS: The total cancer detection rate at prevalent screens was 6.0/1000, 18% being in situ cancers; the detection rate of invasive cancers < or = 10 mm in diameter was 1.3/1000, but data on size were missing for 12% of cancers. Referral rates were significantly lower for programmes using two view mammography at the prevalent screen than for those using single view, and cancer detection rates were significantly higher. For prevalent screens over the three year period, 70% of programmes had a referral rate of < or = 7%, 87% had a benign biopsy rate of < or = 5/1000, and 79% had a cancer detection rate of > or = 5/1000. By contrast, only 30% of programmes appeared to meet the target detection rate of > 1.5/1000 for invasive cancers < or = 10 mm in diameter. CONCLUSIONS: While the majority of interim targets are being met by the NHSBSP, the rate of detection of small invasive cancers requires careful monitoring. Collection of more accurate data on size of cancers and interval cancer rates will give a better indication of progress towards the target mortality reduction. PMID- 8719147 TI - Measurement of the costs in two mammographic screening programmes in the province of Florence, Italy. AB - OBJECTIVES: To estimate and to compare the cost per woman examined and per breast cancer detected in two mammographic screening programmes in the province of Florence. SETTING: Two ongoing, population based, mammographic screening programmes in the province of Florence. The first (district project) was started in the seventies in a rural area, whereas the second (city project) was started in the city at the end of 1990. METHODS: All relevant resources consumed by the programmes (costs) were listed and measured. The costs are related to 1993. The unit cost for each phase (recruitment, screening, assessment) of the screening process was estimated by dividing the total cost of the phase by the number of women examined. The cost per cancer detected was obtained by dividing the total cost of the programme by the number of cancers detected at screening. RESULTS: The costs per woman examined were $38.1 and $41.1 in the district and city programmes respectively. The cost per breast cancer detected was $7424 in the district programme and $5180 in the city programme. Staff accounted for more than 50% of the total cost. The unit cost in the screening phase was higher in the district programme, whereas it was lower in the assessment phase. CONCLUSIONS: Our results are consistent with, although slightly lower than, published estimates of the cost per woman screened. Important parameters for determining the efficiency of a mammographic screening programme are the compliance rate and the recall rate. PMID- 8719149 TI - Evaluating the national screening programme for congenital dislocation of the hip. AB - In 1969 universal clinical screening for neonatal hip instability was formally adopted in the United Kingdom with the aim of detecting and treating children considered to be at high risk of congenital dislocation of the hip (CDH). However, clinical screening is associated with both false positive and false negative diagnoses and has never been evaluated in a randomised trial. The emergence of hip ultrasound provides renewed impetus to reconsider and formally evaluate screening for CDH. Ultrasound imaging of the newborn hip may be used as a screening test and to assess and manage infants with clinically detected hip instability. Universal primary ultrasound screening has been adopted in some European countries, but enthusiasm for this new technology has been tempered by the subsequent large increases in treatment and follow up. This paper reviews the existing evidence to support the different approaches to screening and describes the research agenda of the MRC working party on congenital dislocation of the hip. A randomised trial of screening is required to evaluate the policy options before ultrasound screening becomes widely adopted within the United Kingdom. The feasibility and acceptability of a trial need to be explored and key issues relating to trial design addressed. PMID- 8719148 TI - Compliance after 17 years of breast cancer screening. AB - OBJECTIVE: To investigate changes in attendance over the course of a population based breast screening programme. DESIGN: Longitudinal; analysis of registered data over nine screening rounds. SETTING: Subjects belong to the screening population of the Nijmegen experimental breast screening programme (1975-1990). SUBJECTS: A cohort of 6898 women who were enrolled in 1975 at the age of 50-69 years and who were invited to nine subsequent screening rounds, irrespective of their attendance at previous rounds. RESULTS: Attendance of women aged 50-53 years at entry was high at the initial screening (88%), decreased in the course of the programme, but remained well over 60% until round 8; 39% completed nine rounds and 24% completed seven to eight rounds. Attendance of women who were older at entry was somewhat lower at the initial screening (87%-82%) and declined more strongly. Regular compliance was also lower. Specific compliance patterns are found that can be relevant for further research. CONCLUSIONS: A substantial proportion of eligible women can be committed to mammography every two years, possibly even after reaching the age of 70, if the age at entry is around 50 and the screening programme is well established in the community. Starting a screening programme in older women seems ineffective. PMID- 8719150 TI - Screening for Down's syndrome: the first two years experience in Bristol. AB - OBJECTIVES: To evaluate the effectiveness of a programme for antenatal screening for Down's syndrome using alpha fetoprotein and total human chorionic gonadotrophin as maternal serum markers. SETTING: A district general hospital providing a screening service to a local purchasing authority and (under contract) to another purchasing authority in the same region. METHODS: Patients were counselled and screened between 15 and 20 weeks gestation and Down's risk estimates calculated using the maternal serum marker results as modifiers of the age related risk. Outcome was determined in collaboration with the Regional Cytogenetics Unit. OUTCOME MEASURES: Detection rate for Down's syndrome, false positive rate, uptake of screening, and uptake of amniocentesis. RESULTS: In two years 22816 women were screened (approximately 84% of population); 32 Down's pregnancies were identified, 19 (59.4%) had a reported risk of > or = 1:250 and 20 (62.5%) a reported risk of > or = 1:300. Of those screened before 17 weeks, 16/20 (80%) had a reported risk of > or = 1: 300 compared with 4/12 (33%) of those screened later (P = 0.008); 4.64% of patients screened had reported risks > or = 1: 250 and 5.87% reported risks of > or = 1:300. Amniocentesis uptake was 70% in patients with reported risks of > or = 1:300. CONCLUSIONS: Overall the screening programme was effective but screening before 17 weeks was very much more effective than screening later. PMID- 8719151 TI - A randomised controlled trial of strategies to prompt attendance for a Pap smear. AB - OBJECTIVE: To assess the comparative efficacy, by randomised controlled trial, of three interventions designed to encourage "at risk" women to have a Pap smear: an educational pamphlet; letters inviting attendance at a women's health clinic; and letters from physicians. METHODS: Subjects at risk for cervical cancer who had not been adequately screened were identified by a random community survey and randomly allocated to one of the intervention groups or a control group. Six months after intervention implementation, a follow up survey assessed subsequent screening attendance. Self report was validated by comparison with a national screening data base. RESULTS: A significantly greater proportion of women (36.9%) within the group receiving a physician letter reported screening at follow up than in any other group (P = 0.012). The variables most strongly predicting screening attendance were: age, perceived frequency of screening required, use of oral contraceptives, and allocation to receive the physician letter intervention. CONCLUSIONS: The relative efficacy of the GP letter in prompting screening attendance shows that this strategy is worthy of further investigation. There remains a need to examine the barriers to screening for older women, and to develop tailored strategies for this population. PMID- 8719152 TI - Problems in assessing screening experience in observational studies of screening efficacy: example of urinalysis screening for bladder cancer. AB - OBJECTIVE: To evaluate screening urinalysis for its efficacy in preventing fatal bladder cancer. METHODS: A case-control study of 290 case subjects with fatal bladder cancer upsilon 290 age and sex matched control subjects, with ascertainment of urinalyses by medical record review. RESULTS: In the five years before the symptoms that led to the detection of bladder cancer, case subjects had significantly fewer routine (screening) urinalyses than controls (matched odds ratio for one or more urinalyses upsilon none = 0.60 [95% confidence interval (CI) 0.41 to 0.87], adjusted for cigarette smoking and occupational bladder cancer risk). However urinalyses for urinary symptoms and problems before those that led to bladder cancer diagnosis were more frequent in the cases; hence there was only a small statistically non-significant deficit in cases of urinalyses for all reasons combined (corresponding adjusted odds ratio 0.94, 95% CI 0.61 to 1.46). Since urinalyses for symptoms and problems could have led to the detection of bladder cancer, the latter finding appears more applicable. Methodological problems included: difficulty in assigning the reason for urinalysis; the possibility that clinically indicated urinalyses led to a reduction in screening urinalyses; and difficulty in identifying the detectable and curable preclinical phase for targeting ascertainment of urinalyses. Reducing potential efficacy, haematuria on urinalysis, particularly transient or mild asymptomatic microhaematuria, was often not followed up by cystoscopy. CONCLUSIONS: This study did not show that ordinary urinalysis was effective in preventing fatal bladder cancer but because of methodological problems a clear answer may require a randomised controlled trial. PMID- 8719153 TI - Public understanding of medical screening. AB - AIM: To estimate the extent of public understanding of mass screening for disease and of the benefits and limitations of screening. METHODS: Telephone interviews were conducted with a national sample of 835 Australians. RESULTS: Of total respondents, 68% claimed to have heard of screening tests, but only 21% correctly understood that screening tests are for asymptomatic people. This understanding was related to education. The most frequently named tests were mammography (51%) and Pap tests (33%), and for all age groups these were mentioned by more women than men. Around 27% of respondents thought that the Pap test would detect 95% or more of case, 45% thought the test would detect 90% or more, while 60% of respondents thought the test would detect 80% or more of cases. Around 29% of respondents thought that mammography would detect 95% or more of cases, 49% thought the test would detect 90% or more, while 65% of respondents thought the test would detect 80% or more of cases. Of all respondents, 33% favoured compensation when cases were "missed" by screening provided people were warned beforehand, 58% were not in favour, and 9% were unsure. CONCLUSIONS: There are misconceptions among the public concerning the purpose of screening and the accuracy of screening tests. However, most people accept that some cases of disease will be missed by screening and that if people are adequately informed beforehand compensation should not automatically follow for those whose disease is missed. PMID- 8719154 TI - Detection of pregnancies with trisomy 18 in screening programmes for Down's syndrome. PMID- 8719155 TI - Molecular cloning of a gene from Plasmodium falciparum that codes for a protein sharing motifs found in adhesive molecules from mammals and plasmodia. AB - Adhesion of Plasmodium to host cells is an important phenomenon in parasite invasion and in malaria-associated pathology. We report here the molecular cloning of a putative adhesive molecule from P. falciparum that shares both sequence and structural similarities with a sporozoite surface molecule from Plasmodium termed the thrombospondin-related anonymous protein (TRAP) and, to a lesser extent, with the circumsporozoite (CS) protein. The gene, which is present on chromosome 3 as a single copy, was termed CTRP for CS protein-TRAP-related protein. The full-length CTRP encodes a protein containing a putative signal sequence followed by a long extracellular region of 1990 amino acids, a transmembrane domain, and a short cytoplasmic segment. The putative extracellular region of CTRP is defined by two separated adhesive domains. The first domain contains six 210-amino acid-long homologous repeats, the sequence of which is related to the A-type domain found in adhesive molecules including the alpha subunits of several integrins and a number of extracellular matrix glycoproteins. The second domain contains seven repeats of 87-60 amino acids in length, which share similarities with the thrombospondin type 1 domain found in a variety of adhesive molecules. Finally, CTRP also contains consensus motifs found in the superfamily of haematopoietin receptors. Interstrain analysis of eight different parasite isolates revealed that CTRP does not show size polymorphism except in repetitive regions flanking potential adhesive domains. PMID- 8719156 TI - COS cell expression cloning of Pfg377, a Plasmodium falciparum gametocyte antigen associated with osmiophilic bodies. AB - We report the deduced protein sequence and preliminary characterization of Pfg377, a novel sexual stage antigen of Plasmodium falciparum. An initial cDNA clone (Pfg377-1) encoding the N-terminal 755 amino acids of Pfg377 was isolated by transfecting a 3D7 gametocyte cDNA library into COS7 cells and selecting using a pool of anti-Pfs230 monoclonal antibodies. The protein encoded by Pfg377-1 included an N-terminal hydrophobic signal sequence, but no apparent transmembrane anchor. Instead, the particular cDNA clone selected was fused in-frame at its 3' end with the coding sequence for the human decay acceleration factor membrane anchor, which had been deliberately placed downstream of the vector polylinker in order to attach potential fusion proteins onto the COS cell surface. Northern blots probed with the Pfg377-1 cDNA demonstrated cross-hybridization to a single approximately 9.5-kb transcript, which was present only in sexual stages, and not in a sexual stages. DNA hybridization was used to obtain a series of overlapping genomic clones which collectively yielded the complete DNA sequence for Pfg377. There are no introns within the gene, which contains a 9360-bp open reading frame and encodes a 377-kDa protein. The Pfg377 protein is highly hydrophilic, and has an essentially non-repetitive structure, with only four very limited regions of tandem repeats. The Pfg377 gene resides on chromosome 12, and immunoelectron microscopy with two different anti-Pfg377 polyclonal antisera raised against two separate recombinant sub-fragments of the protein both indicated that the antigen is located in electron-dense organelles of the gametocytes--the osmiophilic bodies--which are proposed to play a role in parasite emergence from the erythrocyte during gametocyte maturation in the Anopheles mosquito midgut. Although it was selected with anti-Pfs230 antibodies, comparison of the sub cellular locations and protein sequences of Pfg377 and Pfs2 show them to be completely distinct antigens. We hypothesize that Pfg377-1 was initially isolated because it expresses an epitope which is recognized by (i.e., cross-reacts with) one of the anti-Pfs230 monoclonal antibodies used to select the original transfected COS cells. PMID- 8719157 TI - Analysis of pfmdr1 and drug susceptibility in fresh isolates of Plasmodium falciparum from subsaharan Africa. AB - Resistance of Plasmodium falciparum to many therapeutic agents is an increasing problem in most endemic areas. The role of the mdr-like gene products of P. falciparum in resistance to quinoline-containing compounds is not clear. The purpose of this study was to further examine the role of pfmdr1 in drug resistance in fresh clinical isolates originating from Africa. Drug susceptibility testing (chloroquine, mefloquine, halofantrine and quinine) and a molecular analysis of pfmdr1 was completed for 51 fresh clinical isolates. A statistical association between the chloroquine sensitivity phenotype and an intragenic allele of pfmdr1 was noted at a position, amino acid 86, which was previously associated with chloroquine resistance. There was little variation in the other intragenic alleles previously associated with chloroquine resistance. No correlation between pfmdr1 intragenic allelic variation and susceptibility to mefloquine, halofantrine or quinine was found. There was no association between gene copy number of pfmdr1 and any drug resistant phenotype in an analysis of selected isolates. This, along with other data, suggests that mefloquine resistance may have arisen by two different mechanisms in African and Southeast Asian isolates. Much more variability in the polyasparaginated region of the pfmdr1 gene was noted in this study than previously reported. In addition, fingerprint analysis using multiplex PCR revealed considerable genetic variability among these isolates. PMID- 8719158 TI - Processing and transport of a lysosomal membrane glycoprotein is developmentally regulated in African trypanosomes. AB - We have used pulse-chase immunoprecipitations methods to study early post translational processing of CBI-gp, a lysosomal membrane glycoprotein expressed by African trypanosomes, Rap67, a polyclonal antibody to CBI-gp, immunoprecipitated a 100-kDa glycoprotein, gp100, from both bloodstream forms (BF) and procyclic forms (PF) of Trypanosoma brucei gambiense immediately after a 5-min pulse with radiomethionine. N-Glycanase digestion released a 67-kDa core protein, p67, from gp100 of both life cycle forms V8 protease digestion of p67 from BF and PF yielded 13 identical methionyl peptides, suggesting that gp100 from both life cycle forms have very similar or identical p67 core molecules. In BF, gp 100 carried both endoglycosidase H (EndoH)-resistant and EndoH-sensitive, N-linked oligosaccharides immediately after labeling. In PF, all the N-linked sugars on gp100 were EndoH sensitive. In BF, gp100 chased progressively into slower migrating 150-180-kDa components that obtained the CBI epitope, traveled to the cell surface where they could be biotinylated, and were proteolytically processed. The increase in mass of gp100 during chase in BF resulted from an elongation of N-linked oligosaccharides. Maturation of gp100 into 150-180-kDa CBI gp was inhibited if BF were chased in the presence of glucosidase inhibitors castanospermine or deoxynojirimycin. In PF, gp100 did not increase in mass, could not be biotinylated on the cell surface, and was not proetolyzed during extended chases. Cryoimmunoelectron microscopy revealed that the antigens detected by rap67 are abundant in lysosomes and endosomes in both BF and PF. Thus, BF and PF express very similar or identical lysosomal membrane glycoproteins but process and transport them in very different ways. PMID- 8719159 TI - Ouabain-sensitive Na+,K(+)-ATPase in the plasma membrane of Leishmania mexicana. AB - The mechanism responsible for the regulation of intracellular Na+ and K+ concentrations in trypanosomatids is unknown. In higher eukaryotes a ouabain sensitive Na+,K(+)-ATPase located in the plasma membrane is the main mechanism for the regulation of the intracellular concentrations of Na+ and K+, while in trypanosomatids there are conflicting evidences about the existence of this type of ATPase. By the use of a highly enriched plasma membrane fraction, we showed that an ouabain-sensitive Na+,K(+)-ATPase is present in L. mexicana. The affinity of the enzyme for Na+ and K+ is similar to that reported for the mammalian Na+,K(+)-ATPase, showing also the same kinetic parameters regarding the relative concentration of those cations that give the optimal activity. Vanadate (10 microM) fully inhibits the ATPase activity, suggesting that the enzyme belongs to the P-type family of ionic pumps. The enzyme is sensitive to ouabain and other cardiac glycosides. These cardiac glycosides do not show any appreciable effect on the higher Mg(2+)-ATPase activity present in the same preparation. By the use of [3H]ouabain, we also show in this report that the binding of the inhibitor to the enzyme was specific. Taken together, these results demonstrate that an ouabain-sensitive Na+,K(+)-ATPase is present in the plasma membrane of Leishmania mexicana. Therefore, this Na+,K(+)-ATPase should participate in the intracellular regulation of these cations in Leishmania. PMID- 8719160 TI - The Leishmania promastigote surface antigen 2 complex is differentially expressed during the parasite life cycle. AB - The promastigote surface antigen 2 (PSA-2) complex comprises a family of antigenically similar polypeptides of M(r) 96,000, 80,000 and 50,000, anchored to the membrane with glycosylphosphatidylinositol. Although PSA-2 was initially detected only in promastigotes, Northern blot analysis indicated that mRNA transcripts are also present in amastigotes. Unlike the situation in promastigotes, where at least four major transcripts (2.6-5.3 kb) were detected, only one major (2.6 kb) and two minor transcripts were present in amastigotes. A cDNA clone encoding a member of the PSA-2 family expressed in amastigotes was isolated using DNA probes. The predicted protein sequence of M(r) 40,000 is distinct from promastigote sequences, but shows significant similarity to previously described members of the family from L major and L amazonensis. Antibodies to the carboxyl terminal sequence conserved in all L major PSA-2 studied to date, as well as antibodies affinity purified on the amastigote cDNA derived polypeptide recognized a major M(r) 50,000 amastigote polypeptide. Immuno electron microscopy localized both promastigote and amastigote PSA-2 to the cell surface. The expression of PSA-2 polypeptides during the transformation of amastigotes into promastigotes was ordered in a time-dependent manner, with the promastigote M(r) 80000 polypeptide appearing first, followed by the M(r) 96000 polypeptide. In contrast to the glycosylphosphatidylinositol anchor of promastigote PSA-2, which could be hydrolysed by phosphatidylinositol-specific phospholipase C, the amastigote form was resistant to this enzyme. PMID- 8719161 TI - Phenotype and genotype diversity in the circumsporozoite proteins of Plasmodium vivax in Thailand. AB - Two phenotypes of the circumsporozoite (CS) protein of the human malaria parasite Plasmodium vivax occur in Thailand, each of which has a characteristic nonamer repeat: GDRA(A/D)GQPA for VK210-type and ANGAG-NQPG for VK247-type. We have sequenced the repetitive domains and flanking regions from 17 specimens collected from a small area, some of which had given ambiguous results in allele-specific hybridization or enzyme-linked immunosorbent assays (ELISA). Base substitutions occurred in non-random, limited patterns that suggest the dissemination of mutations by both unequal crossing-over and gene conversion; most substitutions were silent and phenotypic variation was relatively minor. Sequence variation and number of repeat units were much more variable in VK210-type clones than in those of VK247-type. Each VK210-type isolate with a poor ELISA response contained at least one clone with one of five residue substitutions not found in normally responsive isolates. The absence of obvious hybrid sequences between the two alleles suggests that most successful recombination may have been between sister chromatids, and the limited phenotypic variation suggests that CS antibody does not exert selective pressure on evolution. PMID- 8719162 TI - Absence of the glutamic acid/alanine-rich protein (GARP) genes in the Nannomonas species Trypanosoma simiae and T. godfreyi. PMID- 8719163 TI - Sequence and immunogenicity of the 23-kDa transmembrane antigen of Schistosoma haematobium. PMID- 8719164 TI - Molecular evidence for a close relative of the arthropod endosymbiont Wolbachia in a filarial worm. PMID- 8719165 TI - Evidence for the smallest nuclear genome (2.9 Mb) in the microsporidium Encephalitozoon cuniculi. PMID- 8719166 TI - Sequence and structural characterization of the spliced leader genes and transcripts in Phytomonas. PMID- 8719167 TI - The structural gene for carbamoyl phosphate synthetase from the protozoan parasite Babesia bovis. PMID- 8719168 TI - Molecular cloning and characterization of an Echinococcus multilocularis and Echinococcus granulosus stress protein homologous to the mammalian 78 kDa glucose regulated protein. PMID- 8719169 TI - Effects of lead on systolic and diastolic cardiac functions. AB - In this paper, both systolic and diastolic cardiac functions were evaluated in 54 lead exposed and 24 non-exposed workers by Doppler echocardiography. With regard to systolic cardiac function, the results suggested that cardiac systolic function increased in exposed groups as a compensatory response for the effect of lead on myocardium. To study left ventricular diastolic function, 2.5 MHz pulsed Doppler analyses of transmitral flow velocity were performed from apical four chamber view. The results showed that time-related parameters were comparable among all groups, but blood flow velocity through the mitral valve and Doppler area fractions changed significantly in lead-exposed groups as evidenced by increased value A, decreased value E and E/A ratio. The decrease of diastolic cardiac function was more significant in lead intoxication group. It was also observed in this study that the activity in serum of the MB isoenzyme of creatine phosphokinase (CPK-MB), one of the indices of myocardial damage, was significantly higher in exposed group than that in control (P < 0.05), and a positive correlation was found between CPK-MB activity and Pb-B. It denoted that the increasing of lead burden leads to more release of CPK-MB from the myocardial cells and suggested the existence of slight myocardial damage, which, conceivably, might cause harm to diastolic cardiac function. PMID- 8719170 TI - Intratracheally administered liposomal alpha-tocopherol protects the lung against long-term toxic effects of paraquat. AB - Paraquat is a broad-spectrum herbicide known to produce lung injury via oxidative stress-mediated mechanisms. Different pharmacological strategies have been explored to reduce the formation of these reactive oxygen species and/or prevent their toxic effects in the treatment of paraquat poisoning. The present study was carried out to investigate whether the antioxidant alpha-tocopherol, incorporated into liposomes and delivered directly to the lungs of rats, could protect the organ against the long-term toxic effects of paraquat. Plain liposomes (composed of dipalmitoylphosphatidylcholine, DPPC) or alpha-tocopherol liposomes (8 mg alpha-tocopherol/kg body weight) were administered intratracheally to animals 24 h prior to an intraperitoneal injection of paraquat dichloride (20 mg/kg) and rats were killed 0, 1, 4, 6, 8, 10, 12, 16, 19 or 24 days after paraquat treatment. Results of this study showed that lungs of animals treated with paraquat were extensively damaged, as evidenced by significant increases in lung weight and decreases in lung angiotensin converting enzyme (ACE) and alkaline phosphatase enzyme (AKP) activities. Moreover, paraquat treatment: resulted in a significant reduction in the number of neutrophils in the blood of rats with a concurrent increase in the pulmonary myeloperoxidase activity, suggestive of neutrophil infiltration in the lungs of treated animals. Pretreatment of rats with liposomes alone did not significantly alter the paraquat-induced changes of all parameters examined. On the other hand, pretreatment of rats with alpha tocopherol liposomes, 24 h prior to paraquat challenge, attenuated paraquat induced changes in ACE, AKP and myeloperoxidase activities but failed to prevent increases in lung weight. Thus, pretreatment of rats with liposome-associated alpha-tocopherol appears to protect the lung against some of the toxic effects of paraquat. PMID- 8719171 TI - Socio-economic, environmental and health aspects of farm workers engaged in mango plantations. AB - A cross-sectional survey of 489 male subjects in the age group 15 to 65 years engaged directly or indirectly in mango cultivation along with 208 control subjects was carried out to find their socio-economic, environmental and health conditions. The conditions like high illiteracy rate (49.5%), poverty (PCI less than Rs 100 per month, 52.2%), poor housing (mud houses, 66.7%) unsafe water supply (78.6%) were prevailing in the surveyed population. The high respiratory morbidity may be attributed to high prevalence of smoking and prolonged inhalation of organic dusts during farming operation associated with illiteracy and poor socio-economic status. Gastrointestinal disorders were related to poor hygienic conditions, smoking and consumption of contaminated water. The symptoms pertaining to CNS, skin and eyes were found to be associated with exposure to pesticides. PMID- 8719172 TI - Harmful effects of MSG on function of hypothalamus-pituitary-target gland system. AB - It has been demonstrated that neonatal administration of monosodium glutamate (MSG) results in a clearly defined lesion of the arcuate nucleus (AN) of the hypothalamus. The present study shows that fat was accumulated in the abdomen of male rats treated with MSG; weights of the body, pituitary and testis were lower; beta-EP content in hypothalamus decreased while L.EnK content increased; serum LH, FSH, TSH, GH and TS levels all decreased in varying degrees while serum PRL level significantly increased. The cAMP content lowered in pituitary, but nor in testes; clear histological changes occurred in testicular tissue; Se-GSH-Px activity in both testis and adrenal gland lowered while LPO level significantly increased. Both Se-GSH-Px activity and LPO level in liver increased. These results indicate that MSG is harmful to the function of the hypothalamus pituitary-target system of neonatal rats. PMID- 8719173 TI - Recommended diagnostic criteria for occupational chronic lead poisoning. AB - The present study aims to recommend the normal upper limit, the acceptable upper limit, the subclinical lead absorption and intoxication diagnostic criteria in an effort to re-evaluate the current national diagnostic criteria for the occupational chronic lead poisoning. The study was conducted on 330 lead exposed workers and 100 non-exposed controls based on the determination of blood and urinary lead, porphyrin metabolism indices, as well as other indices under a nationwide quality control program. The data were subsequently treated by the curve fitting, multi-step transformation to Gauss distribution, and discriminant analysis with the aid of a SAS software package. The relationships between the air lead and blood lead level with certain biological parameters indicative of excessive lead exposure and poisoning were well established. The sensitivity, specificity, accuracy, false positive and false negative results of these critical values were also fully evaluated. This study would be useful for the amendment of the new edition of the national diagnostic criteria for the occupational chronic lead poisoning in China and would provide new approaches for similar investigations. PMID- 8719174 TI - A survey of moniliformin contamination in rice and corn from Keshan disease endemic and non-KSD areas in China. AB - Keshan Disease (KSD) is an endemic heart disease and moniliformin (MF) has been suggested as one of the etiological factors. In this paper, thin layer chromatographic (TLC) and high pressure liquid chromatographic (HPLC) methods were used for the determination of MF in the rice and corn samples collected from KSD areas and non-KSD areas. One hundred and twenty-three rice samples were analyzed and showed MF contamination in only 8 samples (KSD areas: 8.4% positive; non-KSD areas: 2.5% positive) ranging from 73.6 to 265.3 ng/g (mean concentration: KSD areas 156.3 ng/g; non-KSD areas 179.5 ng/g). One hundred and four corn samples in KSD areas and non-KSD areas were determinated by HPLC method, 45.2% samples were contaminated with MF (KSD areas: 81.4%; non-KSD areas: 19.7%) ranging from 52.3 to 1116.0 ng/g (mean concentration: KSD areas 488.9 ng/g; non-KSD areas 457.4 ng/g). The results showed that the contamination of MF in grains were significantly different between rice and corn, but not between the grains from the KSD areas and non-KSD areas, then casting doubt on the role of MF as an etiological factor of KS. PMID- 8719175 TI - Evaluation of medical cost lost due to smoking in Chinese cities. AB - Smoking induces substantial diseases on both individual and the whole society. To identify the true smoking-attributable economic loss, we introduce medical cost accounting as a means to calculate disease-specific medical cost, including inpatient and outpatient cost of those diseases caused by smoking. Medical cost is defined as health resource consumption in terms of money. Cost is allocated to department and services according to coefficient of benefit and operation time. The study in 1988 indicates that total smoking-attributable medical cost is 2.32 billion RMB Yuan in China, 1.70 billion RMB Yuan for outpatient, 0.62 billion RMB Yuan for inpatient. If indirect cost is included, the cost will be greater. Chronic obstructive emphysema has the highest proportion (55.41%) in smoking attributable medical cost. PMID- 8719176 TI - An evaluation on smoking-induced health costs in China (1988-1989). AB - A study on smoking-attributable health economic costs in China was conducted from 1988-1992, in which three major categories of chronic diseases, diseases of cancer, diseases of circulatory system, and diseases of respiratory system were included. A prevalence-based method which estimated the cumulative effect of cigarette smoking during the past 20-30 years was used. The results show that in 1989, the total smoking-attributable economic costs to health sectors in China were about 27.1 billion of Chinese Yuan, including about 7 billion Yuan in direct medical costs and 20 billion Yuan in indirect costs, which include indirect morbidity costs and indirect mortality costs. The relatively low direct costs reflected the low medical costs at hospitals in China at that time. And the high proportion of indirect costs relative to the total costs shows the high potential years of life lost due to cigarette smoking. The results also show the heavier health burden in urban areas than in rural areas, reflecting the worse situation in urban China at nowadays. But if considering that almost 80% of the Chinese are rural farmers with the higher smoking prevalence and relatively shorter history of manufactured cigarette smoking than their urban counterparts, the very frightful situation due to cigarette smoking would be for China in the next century. PMID- 8719177 TI - A natural vaccine candidate strain against cholera. AB - E1 Tor Vibrio cholerae (EVC) strains may be classified into two kinds epidemigenic (EEVC) strains and non-epidemigenic (NEEVC) strains-based on a phage biotyping system. A large number of EEVC strains have been screened for toxigenic and putative colonization attributes. One such naturally occurring strains (designated IEM101) has been found which is devoid of genes encoding cholera toxin (CT), accessory cholera enterotoxin (ACE), zonula occludens toxin (ZOT), but possesses RS1 sequences and toxin-coregulated pilus A gene (icpA) although icpA is poorly expressed. It expresses type B pili but does not possess type C pili. It is an E1 Tor Ogawa strain and does not cause fluid accumulation in rabbit ilcal loop tests. Active immunization of rabbits with strain IEM101 elicited good protection against challenge with virulent strains of V. cholerae O1. Oral administration caused no side effects in 15 human volunteers, colonized the gut for four to ten days and elicited good immune responses. PMID- 8719178 TI - In vivo and in vitro cytogenetic effect of supermethrin. AB - The synthetic pyrethroid insecticide supermethrin (RS)-alpha-cyano-3 phenoxybenzyl-(1RS)-cis-3 (2, 2-dichlorvinyl)-2, 2-dimethyl cyklopropane carboxylate, was tested for its clastogenic ability by in vivo and in vitro assays. In vivo assay consisted of chromosome analysis in sheep bone marrow cells after six weeks subchronic oral administration of 200 mg/kg or 300 mg/kg. The results were compared to induction of chromosome aberrations (CA) in mouse bone marrow cells after acute ip administration of supermethrin in the range 1/2 to 1/10 LD50. Small statistically insignificant increases of CA were found up to 1/2 LD50 dose for mice. Cultured sheep peripheral lymphocytes were used for in vitro clastogenicity evaluation. The final concentrations were 6 x 10(-6) mol.L-1, 6 x 10(-5) mol.L-1 and 6 x 10(-4) mol.L-1. No significant differences were detectable in supermethrin's clastogenic potency at concentrations of 6 x 10(-6) mol.L-1 and 6 x 10(-5) mol.L-1 respectively. The highest concentration was positive (P < 0.05), and a decrease of mitotic index (MI) was also observed. Supermethrin was investigated for the induction of sister chromatid exchanges (SCE) in sheep peripheral lymphocytes, but inadequate results of genotoxic potency were found. The tested pyrethroid was characterized as rather toxic than clastogenic or genotoxic. PMID- 8719179 TI - Functional maturation of neuroendocrine gonadal axis is altered by specific phase relations of circadian neurotransmitter activity in Japanese quail. AB - The present study was designed to ascertain the effects of temporal relationship of circadian neural oscillations on puberty attainment and reproductive growth of Japanese Quail, Coturnix coturnix japonica. Serotonin and dopamine precursors (5 hydroxytryptophan, 5-HTP and L-dihydroxyphenylalanine, L-DOPA; 5 mg/100 g body weight) were injected daily, 8 and 12 h apart in two groups of one-day old chicks, while controls received two daily injections of normal saline. Weekly/biweekly observations (body weight, cloacal gland size, testicular volume and activity, ovarian follicular diameter and rate of egg production) were made until 9 weeks of age, when the experiment was terminated. Results indicate that 8 h relationship completely suppressed gonadal growth even under long photoperiod (LD 16:8), while a 12 h relationship induced precocious sexual maturity and increased the rate of reproduction (spermatogenesis and egg production). It is concluded that circadian phase relationship of serotonergic and dopaminergic activity may not only determine the onset of reproduction in this poultry species, but may also alter the rate of reproduction possibly by affecting photoperiodic mechanism of reproductive regulation. PMID- 8719180 TI - A developmental and component analysis of active sleep. AB - A wide variety of hypotheses have been put forth that address the functional significance of active sleep. Despite the well-accepted fact that active sleep expresses itself predominantly in the perinatal period, the vast majority of these functional hypotheses are applicable largely, if not exclusively, to the adult. We build on the developmental approaches of previous researchers and propose that the individual components of active sleep (e.g., myoclonic twitches, rapid eye movements) exhibit unique developmental and phylogenetic histories and may serve independent functions in the developing organism. This dynamic perspective leads to specific experimental approaches aimed at the developmental roles of these components in the neonate, their maintenance roles in the adult, and the means by which these various components coalesce temporally in what is commonly referred to as a behavioral state. PMID- 8719181 TI - A lesion and neuroanatomical tract-tracing analysis of the role of the bed nucleus of the stria terminalis in retrieval behavior and other aspects of maternal responsiveness in rats. AB - The ventral part of the bed nucleus of the stria terminalis forms a junctional region between the medial and lateral preoptic areas. Previous work has shown that the neurons in this region express Fos-like immunoreactivity during maternal behavior, suggesting their involvement in maternal behavior control. Supporting this hypothesis, the first experiment shows that excitotoxic amino acid lesions of the bed nucleus of the stria terminalis disrupt retrieval behavior and other aspects of maternal responsiveness in postpartum rats. The second study traces the efferent projections of the ventral bed nucleus with the anterograde tracer Phaseolis vulgaris leucoagglutinin. The following regions receive strong projections: lateral septum, substantia innominata, paraventricular hypothalamic nucleus, ventral premammillary nucleus, supramammillary nucleus, paraventricular thalamus, ventral tegmental area, periaqueductal gray, retrorubral field, and the region surrounding the locus coeruleus. PMID- 8719182 TI - Auditory brainstem response in sheep. Part II: Postnatal development. AB - The auditory brainstem response (ABR) was recorded from 15 chronically instrumented lambs between birth and 7 weeks of age as well as from four adult ewes. Latency changes observed within two days of birth may be related to the dissipation of fluid from the lamb external auditory canal and middle ear cavity. Developmental changes observed in the lambs included an early period of wave latency decrements from 1 to 4 weeks followed by a subsequent increase in wave latencies from 4 to 7 weeks. These changes were significant for the I-IV interwave interval, possibly reflecting the combined effects of increased myelinization and increased neural track length. Responses from 7 week-old lambs were not significantly different from those obtained from the ewes. PMID- 8719184 TI - Lessons learned. PMID- 8719183 TI - Salicylate-induced changes in auditory thresholds of adolescent and adult rats. AB - Shifts in auditory intensity thresholds after salicylate administration were examined in postweanling and adult pigmented rats at frequencies ranging from 1 to 35 kHz. A total of 132 subjects from both age levels were tested under two-way active avoidance or one-way active avoidance paradigms. Estimated thresholds were inferred from behavioral responses to presentations of descending and ascending series of intensities for each test frequency value. Reliable threshold estimates were found under both avoidance conditioning methods, and compared to controls, subjects at both age levels showed threshold shifts at selective higher frequency values after salicylate injection, and the extent of shifts was related to salicylate dose level. PMID- 8719185 TI - Recovery from mivacurium-induced neuromuscular blockade is not affected by anticonvulsant therapy. AB - Long-term chronic anticonvulsant therapy produces a resistance to the effects of all nondepolarizing neuromuscular blocking agents studied to date. Since the metabolism of mivacurium is unique among the nondepolarizing neuromuscular blocking agents, the effect of anticonvulsants on its recovery parameters was examined. Forty-five patients were separated into three groups based on the number of chronic anticonvulsant medications the subjects were taking: subjects in group 1, the control group, took no anticonvulsant medication; group 2 subjects took one medication; and group 3 subjects took two medications. Mivacurium, 0.15 mg/kg i.v., was administered after induction of general anesthesia with thiopental sodium, 4-6 mg/kg, and fentanyl 2-4 micrograms/kg i.v. Maintenance anesthesia consisted of N2O in O2. 0.2-0.3% end-tidal isoflurane, and a fentanyl infusion. The evoked compound electromyograph (ECEMG) of the adductor pollicis-brevis muscle was measured for time of onset, T-1 (time at which ECEMG signal reaches 5, 25, 50, and 75% of baseline), TR (TOF ratio), and recovery index. T-1 at 25% was 18.2 +/- 1.8, 20.7 +/- 1.9, and 21.5 +/- 1.4 min for groups 1, 2, and 3, respectively, with TR at 25% being 23.7 +/- 2.3, 26.9 +/- 2.4, and 27.3 +/- 2.3 min. No significant differences were noted in neuromuscular recovery between groups at any time point. These results fail to demonstrate the resistance to the nondepolarizing neuromuscular blockade of mivacurium that has been observed with other nondepolarizing agents. PMID- 8719186 TI - The effects of surgical stimulation on intracranial hemodynamics. AB - This study investigates the effects of surgical stimulation on cerebral blood flow velocity using transcranial Doppler sonography (TCD) in 1 and 2 maximum alveolar concentration (MAC) isoflurane anesthetized patients. Sixty ASA I and II patients undergoing breast surgery were studied. Anesthesia was maintained with 0.6% isoflurane (groups 1 and 2) or 1.2% isoflurane (groups 3 and 4) and nitrous oxide in oxygen (FIO2, 0.33). TCD recordings of middle cerebral artery mean blood flow velocity (Vmean, cm/s) were taken before each respective treatment and for the 15-min investigation period. In groups 1 and 3 (each n = 20), the patients were exposed to surgical stimulation (skin incision). In groups 2 and 4 (each n = 10), norepinephrine infusion (0.1 microgram.kg-1.min-1) was used to increase mean arterial blood pressure (MAP) to levels similar to those seen with surgical stimulation (groups 1 or 3). Body temperature and PETCO2 remained constant over time and did not vary between treatment groups. In groups 1 and 3, MAP increased 22 and 16% after surgical stimulation. In groups 2 and 4, MAP increased 28 and 36% after norepinephrine infusion. Vmean was increased 23 and 17% after surgical stimulation during 1 and 2 MAC isoflurane but did not change with norepinephrine infusion. These data show that cerebral blood flow velocity increases with surgical stimulation in 1 and 2 MAC isoflurane-anesthetized patients. This is not a function of changes in MAP. These data suggest that surgical stimulation increases cerebral blood flow, possibly because of arousal. PMID- 8719187 TI - The bispectral index during induction of anesthesia with midazolam and propofol. AB - This study evaluated the bispectral index as an indicator of anesthetic depth in relation to the cardiovascular response to intubation. Two treatments were compared: group 1 (n = 8) received propofol for induction of anesthesia (2 mg/kg bolus followed by an infusion of 0.20 mg/kg-1/min-1, group 2 (n = 8) was given 90 micrograms/kg midazolam 2 min before, followed by anesthesia with half-strength propofol (1 mg/kg bolus with infusion of 0.10 mg/kg-1/min-1). The bispectral index of the electroencephalogram, blood pressure, and heart rate were measured under unanesthetized conditions, during anesthetic induction, intubation, and a 15-min period after intubation. The duration of anesthesia and the total propofol requirement were recorded. Midazolam pretreatment produced transient decreases in blood pressure and the bispectral index. During anesthetic induction with propofol, blood pressure decreased 20% in both groups, and the bispectral index decreased to lower levels in group 1 (29 +/- 9) than in group 2 (47 +/- 22). Intubation increased blood pressure more in group 2 (50 +/- 10 mm Hg) than in group 1 (30 +/- 12 mm Hg). Throughout the rest of the surgery, more propofol was used in group 1 (77 +/- 14 micrograms/kg-1/min-1) than in group 2 (42 +/- 14 micrograms/kg-1/min-1). These results show that the decrease in bispectral index provides an indication of the blood pressure increase to intubation during propofol anesthesia. Midazolam pretreatment did not attenuate the cardiovascular response to intubation but did decrease propofol use during surgery. PMID- 8719188 TI - Limitations of jugular bulb oxyhemoglobin saturation without intracranial pressure monitoring in subarachnoid hemorrhage. AB - We report a case of subarachnoid hemorrhage in which, even after having obtained a normal jugular bulb oxyhemoglobin saturation, cerebrovenous desaturation developed, and brain death occurred. The limitations of jugular bulb oxyhemoglobin saturation without intracranial pressure monitoring are discussed. We conclude that if increased intracranial pressure is suspected, use of jugular bulb oxyhemoglobin saturation monitoring alone would appear to be substantially limited. PMID- 8719189 TI - Delayed presentation of cerebral arterial gas embolism following proven intraoperative venous air embolism. AB - We describe the case of a 33-year-old woman who suffered a delayed-onset arterial gas embolism following a significant venous air embolism during surgery to remove an acoustic neuroma. We report the management of the problem and discuss the mechanisms by which this event might have occurred. PMID- 8719190 TI - Cerebral ischemia after venous air embolism in the absence of intracardiac defects. AB - Cerebral air embolism occurred in a patient undergoing posterior fossa surgery performed in the sitting position for acoustic neuroma removal. The patient experienced two episodes of venous air embolism, as evidenced by precordial Doppler, end-tidal carbon dioxide reduction, and oxygen desaturation. In both cases, air was aspirated from the central venous catheter; during the second episode there was arterial hypotension and electrocardiogram changes, and air bubbles were visualized in the cerebellar arteries. The patient did not regain consciousness after surgery and developed early tonic-clonic convulsions and electroencephalogram status epilepticus, which was treated with barbiturate coma. Intracardiac septal defects were not detected by transesophageal echocardiography, and computerized tomography of the brain demonstrated multifocal discrete ischemic areas in the cerebral hemispheres. The patient died 6 days after surgery without having regained consciousness. This case appears to represent the occurrence of transpulmonary passage of venous air embolism. PMID- 8719191 TI - Ketorolac appears to decrease polyuria during intracranial surgery. AB - Two cases are described of patients, undergoing surgery for intracranial pathology, who developed polyuria intraoperatively. The urine output decreased to within normal range shortly after the administration of ketorolac, a nonsteroidal antiinflammatory drug. The possible mechanisms producing the increased urinary output and the influence of prostaglandins on renal function are discussed. PMID- 8719192 TI - A comparison of the electrophysiologic characteristics of EEG burst-suppression as produced by isoflurane, thiopental, etomidate, and propofol. AB - Electroencephalogram (EEG) burst-suppression can be produced with several anesthetic agents. Discussions of burst-suppression suggest that it has been viewed by many as a relatively uniform physiologic state independent of the agent used to produce it. This view may be an oversimplification. In this study, relatively deep EEG burst-suppression (suppression to burst time ratio, 4:1) was induced in rats with isoflurane (I), thiopental (T), etomidate (E), and propofol (P). Burst duration, maximum peak-to-peak voltage, area under the curve, and the ratio of power in high versus low frequencies of EEG recorded in both cortex and subcortex (thalamus) were determined. Analysis of the bursts revealed significant differences in duration [I, 1.4 +/- 0.4 (SD); T, 0.8 +/- 0.5; E, 0.3 +/- 0.1; P, 0.4 +/- 0.1 seconds], peak-to-peak voltage (I, 488 +/- 146; T, 285 +/- 106; E, 310 +/- 87; P, 249 +/- 50 muV), and area under the curve (I, 111 +/- 24; T, 35 +/ 31; E, 17 +/- 7; P, 21 +/- 4 muV-s) for all agent pairs except etomidate and propofol. Suppression phase analysis revealed considerable residual activity with all four agents, although peak-to-peak voltage (I, 129 +/- 29; T, 64 +/- 20; E, 62 +/- 11; P, 40 +/- 15 muV) and area under the curve (I, 73 +/- 17; T, 37 +/- 14; E, 30 +/- 5; P, 22 +/- 10 muV-s) were greatest with isoflurane. The cortical versus subcortical comparison revealed, for all agents, greater peak-to-peak voltage and area under the curve in the subcortex. The data indicate that the electrophysiologic characteristics of burst-suppression vary among the four agents, with the possible exception of etomidate and propofol. The data suggest that the neurophysiologic states associated with burst-suppression produced by various anesthetics should not be assumed to be uniform. PMID- 8719193 TI - Effect of desflurane anesthesia on transcortical motor evoked potentials. AB - The effect of the volatile anesthetic desflurane on motor evoked potentials was examined in male rats. Animals underwent cortical stimulation using small platinum ball stimulating electrodes secured on the motor cortex. To record evoked compound muscle action potentials (CMAPs), single-shock electrical stimulation was delivered to the forelimb representation of the motor cortex. Muscle responses were readily obtained in the contralateral extensor muscles. The effect of desflurane was examined at various concentrations ranging from 0.7 to 11.4%. With increasing concentrations of desflurane, there was a progressive decrease in the CMAP amplitude and systemic blood pressure over the baseline values. This decrease became statistically significant (p = 0.0078) at 5.7% [1 maximum alveolar concentration (MAC)] concentration of desflurane. Although there was a decrease in heart rate, the results were not statistically significant (p = 0.03). No significant difference in the onset latency or the duration of the CMAP was noted at different concentrations of the anesthetic. We conclude that desflurane anesthesia significantly alters the amplitude of the muscle response evoked by motor cortex stimulation in experimental animals. PMID- 8719194 TI - Suppression of cerebral metabolic rate for oxygen (CMRO2) by mild hypothermia compared with thiopental. AB - If the efficacy of hypothermia and barbiturates in ameliorating ischemic brain injury lies in reducing the cerebral metabolic rate of oxygen (CMRO2), the greater efficacy of mild hypothermia (34 degrees C) compared with barbiturates is inconsistent with the 15-20% reduction of CMRO2 caused by mild hypothermia compared with 50% caused by barbiturates. This paradox, we hypothesized, derives from the fact that whereas barbiturates lower CMRO2 associated with EEG activity or thiopental (TP)-suppressible CMRO2, not essential for cellular viability, hypothermia lowers CMRO2 associated with providing energy, i.e., adenosine triphosphate, to maintain transmembrane ion gradients or TP-nonsuppressible CMRO2, essential for neuronal viability. To test this hypothesis, we measured whole brain cerebral blood flow (CBF) and CMRO2 in two groups of rats mechanically ventilated with 70% N2O/30% O2 before and after TP-induced isoelectric EEG. In the normothermic group (n = 7), measurements were made at a brain temperature (Tb) of 38 degrees C, while in the hypothermic group (n = 7), they were made at 34 degrees C. In the normothermic group, TP-induced isoelectric EEG reduced CMRO2 by 50%, from 7.92 +/- 1.05 to 3.95 +/- 0.70 ml 100 g-1 min-1 (mean +/- = SD). Thus, at 38 degrees C, TP-suppressible and TP-nonsuppressible CMRO2 were both 50 +/- 4% of total CMRO2. In the hypothermic group, decreasing Tb from 38 to 34 degrees C caused a 17% decline in CMRO2, from 7.62 +/- 1.92 to 6.28 +/- 1.22 ml 100 g-1 min-1 (p > 0.05). AT 34 degrees C, TP infusion lowered CMRO2 to 2.15 = 0.46 ml 100 g-1 min-1. At 34 degrees C, TP-suppressible and TP nonsuppressible CMRO2 values were 64 +/- 7% and 36 +/- 8% of total CMRO2, respectively. TP lowered CBF by 50% at both 38 and 34 degrees C. In conclusion, mild hypothermia selectively lowers TP-nonsuppressible CMRO2 associated with the maintenance of viability rather than EEG-associated or TP-suppressible CMRO2. PMID- 8719195 TI - Making clinical decisions based on animal research data: pro. PMID- 8719196 TI - Early application of the results of animal experimentation to human clinical trials: con. PMID- 8719197 TI - Active cooling during anesthesia and critical care. PMID- 8719198 TI - Important aspects in the treatment of severe accidental hypothermia: the Innsbruck experience. AB - The purpose of this paper is to review important aspects in the treatment of accidental hypothermia, based on our own experience in rewarming 55 patients with severe accidental hypothermia and a core temperature < 30 degrees C. We used three different methods of rewarming, adjusted to the patients' hemodynamics: airway rewarming, warmed fluids and insulation in patients with stable hemodynamics (group 1, n = 24), peritoneal dialysis in patients with unstable hemodynamics (group 2, n = 7) and extracorporeal circulation in patients with cardiocirculatory arrest (group 3, n = 24). Survival rates were 100% (group 1), 72% (group 2) and 13% (group 3) retrospectively. Published data supporting our strategy and alternative approaches are reviewed. The method used to rewarm a patient with severe accidental hypothermia should be adjusted to the hemodynamic status. The prognosis is excellent in patients in whom no hypoxic event precedes hypothermia and no serious underlying disease exists. PMID- 8719199 TI - Mild resuscitative hypothermia and outcome after cardiopulmonary resuscitation. AB - Recovery without residual neurological damage after cardiac arrest with global cerebral ischemia is still a rare event. Severe impairment of bodily or cognitive functions is often the result. The individual, emotional, and social aspects of brain damage and rehabilitation are seldom taken into account. Efforts to improve the prevention of brain damage immediately after successful resuscitation of patients are missing. The efficacy of hypothermia in preserving neurologic function when instituted before and during certain no-flow cardiovascular states has been well documented both clinically and experimentally since the 1950s. Most studies have used moderate (28-33 degrees C) to deep (20-28 degrees C) hypothermia to demonstrate these protective effects. Considering the use of hypothermia for preservation and resuscitation, the lack of controlled outcome trials, the long period of time required to reach therapeutic hypothermia, and the incidence of rewarming complications such as infection, arrhythmia, and coagulopathy have made it difficult to apply these methods to emergency situations such as cardiac arrest. Recent experimental evidence in dogs has shown that hypothermia induced after cardiac arrest does indeed mitigate the effects of the postresuscitation syndrome and improves neurologic function and reduces histologic brain damage. More importantly, such benefits can be demonstrated with mild (34-36 degrees C) hypothermia, thus minimizing complications and requiring less time for induction of hypothermia. Ice water nasal lavage, direct carotid infusion of cold fluids, use of a cooling helmet, and peritoneal cooling are promising techniques for clinical cerebral cooling. External auditory canal temperature (e.g., tympanic membrane temperature changes) could provide an approximation to brain temperatures. For accurate temperature monitoring, however, a central pulmonary artery thermistor probe should be inserted. Temperature monitoring is needed to avoid temperature < 30 degrees C. Mild hypothermia may prove to be an important and secure component for cerebral preservation and resuscitation during and after global ischemia; it may also prove to be a useful method of cerebral resuscitation after global ischemic states, thereby promoting the prevention of neuromental diseases. PMID- 8719200 TI - Histopathology and epidemiology of condemnations due to squamous cell carcinoma in broiler chickens in North Carolina. AB - Squamous cell carcinoma (SCC) lesions were confirmed by histopathology in 94% of 176 carcasses condemned for SCC during 8-hour shifts at four different processing plants. Lesions in the remaining carcasses were diagnosed as focal ulcerative dermatitis. SCC lesions were confined to the skin; no metastasis or invasion of underlying muscle was present. Monthly prevalence of SCC condemnations at processing was obtained from two plants for a 1-year period and from two additional plants for 3-year periods. Overall study prevalence of SCC varied between 0.03% and 0.09% of total condemnations. Servicemen were found not to have an effect on SCC condemnations for the only company that had these data available. Chronological trends were evaluated for SCC condemnations. A trigonometric model predicted 6-month cycles for one processing plant over a 3 year period. The same model was used to describe 6-month cycles present in the last 12 months of a 3-year observation period in a different processing plant. Chronologic trends were evaluated qualitatively for the two other processing plants. PMID- 8719201 TI - Determination of optimum formulation of a novel infectious bursal disease virus (IBDV) vaccine constructed by mixing bursal disease antibody with IBDV. AB - A novel vaccine against infectious bursal disease virus (IBDV) has been developed. The new vaccine was constructed by mixing bursal disease antibody (BDA) contained in whole antiserum with live IBDV before lyophilization. To establish various formulations of BDA and IBDV, several BDA doses between 5 units and 80 units of BDA/50 microliters were mixed with 100 EID50/50 microliters of IBDV suspension in Expt. 1; in Expt. 2, several IBDV doses between 10 EID50/50 microliters and 977 EID50/50 microliters of IBDV suspension were mixed with 24 units of BDA/50 microliters. Vaccine preparations were administered subcutaneously to the nape of 1-day-old specific-pathogen-free (SPF) chicks. Safety, potency, and immunogenicity of the different vaccine formulations were evaluated using bursal weight, bursal gross examination, and IBDV antibody titer. Some bursae were examined histologically to confirm gross examinations. Several vaccine formulations were safe and efficacious and met the safety, potency, and immunogenicity criteria. A vaccine construct of 100 EID50 mixed with 24 units of BDA was selected as the release dose. When administered at 1 day of age, the novel vaccine allows for delayed infection of the bursa until after days 6-8 of age in SPF chicks, while initiating potency and immunogenicity to an IBDV challenge. The addition of BDA to the IBDV results in a complex vaccine that allows for safer immunization in SPF birds than under administration of the vaccine virus without BDA. PMID- 8719202 TI - Failure of maternally derived yolk IgG to reach detectable concentrations in the sera of nestling budgerigars (Melopsittacus undulatus). AB - Transfer of maternal immunoglobulin G (IgG) to the yolk and nestling was investigated in the budgerigar. Specific antibodies to avian polyomavirus and Newcastle disease virus could be detected in 82% of yolk extracts of eggs from seropositive hens. Using a double immunodiffusion assay with anti-chicken IgG antibodies, IgG could also be detected in yolk supernatants with virus neutralizing activity. In all assays, IgG concentrations in the yolk extracts were significantly less than those of the adult budgerigar serum. No antiviral activity was detected in nestling serum. Examination of nestling serum with the double immunodiffusion assay and an immuno-dot-blot technique specific for IgG showed that detectable concentrations of IgG are not present in nestling serum until after the yolk sac is fully absorbed. This observation, coupled with the absence of specific anti-viral antibody in nestlings of seropositive hens, indicated that none of the yolk sac antibody reached the nestling circulation. PMID- 8719203 TI - Eimeria tenella and E. acervulina: differences in ability to elicit cross-species protection as compared with the turkey coccidium, E. adenoeides. AB - Repeated oral inoculation of turkey poults with large doses (1 x 10(6) oocysts) of the chicken coccidia, Eimeria tenella or E. acervulina, failed to prevent weight loss, poor feed conversion, and intestinal pathology in turkeys challenged with the turkey coccidium, E. adenoeides. Invasion by E. tenella in turkeys was significantly greater than invasion by E. adenoeides in chickens; by 24 hr postinoculation (PI), the numbers of E. tenella and E. adenoeides sporozoites in the ceca had decreased markedly as compared with the numbers that initially invaded, and they did not differ significantly from each other. At 24 hr PI, however, transfer of cecal scrapings from chickens or turkeys inoculated with E. adenoeides produced infection in 53% of the recipient turkeys, but transfer of scrapings from either chickens or turkeys inoculated with E. tenella failed to produce infection in 20 attempts with recipient chickens. Cultured chicken peripheral blood monocytes (PBMs) that were inoculated with E. adenoeides sporozoites contained numerous vesicles that were recognized by the refractile body-specific monoclonal antibody 1209; the number of vesicles was markedly decreased in PBM cultures inoculated with gamma-irradiated E. adenoeides sporozoites. Very few vesicles were detected in the cytoplasm of turkey PBMs that contained E. tenella sporozoites, and none were detected in turkey PBMs containing E. adenoeides sporozoites. The survival of infective sporozoites, along with the secretion of refractile body antigen, may be more critical to the development of cross-species immunity than the number of sporozoites that initially invade the foreign host. PMID- 8719204 TI - In vitro studies of Campylobacter jejuni/coli strains from hens and humans regarding adherence, invasiveness, and toxigenicity. AB - Campylobacter jejuni/coli strains from hens and humans were compared for their ability to adhere to and invade HEp-2-cells and for toxigenicity to CHO-cells. In both hen and human strains, invasiveness was higher among non-toxigenic strains than among toxigenic ones. The frequency of adherence, invasiveness, and toxigenicity was the same in hen and human strains. PMID- 8719205 TI - Immunosuppressive potential and pathogenicity of an avian adenovirus isolate involved in hydropericardium syndrome in broilers. AB - The role of avian adenovirus isolate PARC-1 as an immunosuppressive agent was investigated using a Newcastle disease virus (NDV) vaccine immune response procedure. The immunosuppressive effect on the humoral immune response was investigated up to 21 days after inoculation with adenovirus. Infected chickens showed a serologic response to NDV that was reduced compared with that of the controls. To further investigate the effect of the virus on major lymphoid organs, the pattern of virus dissemination in various organs was studied at various time intervals after inoculation. Spleen, thymus, bursa of Fabricius, and cecal tonsils of broilers were examined using a dot-immunobinding assay. The virus was found to have a predilection for lymphoid organs, and virus from lymphoid organs was capable of producing disease when inoculated into healthy chickens. The relationship of virus predilection to its immunosuppressive effect also was studied. PMID- 8719206 TI - Application of polymerase chain reaction with arbitrary primers to strain identification of Mycoplasma gallisepticum. AB - DNA heterogeneity among strains and isolates of Mycoplasma gallisepticum (MG) was demonstrated with the arbitrarily primed polymerase chain reaction (AP-PCR) method. This method involves three cycles of low-stringency amplification followed by PCR at higher stringency. Reproducible DNA fragments of 25 different MG strains or isolates were generated with three arbitrarily chosen primers. The MG strains or isolates were distinguished according to the banding patterns of their amplified DNA on agarose gels, and the differences were characteristic for specific isolates. This method is rapid, simple, and reproducible, and it can also be used to determine the similarity between isolates of MG from various sources. PMID- 8719207 TI - B cells and T-lymphocyte subsets of the head-associated lymphoid tissues of the chicken. AB - The head-associated lymphoid tissues of the chicken, composed of the harderian gland and the conjunctiva-associated lymphoid tissue (CALT), were studied to determine whether changes occurred in lymphocyte subpopulations as chickens age from 1 week to 8 weeks. The B cells and subpopulations of T-lymphocytes of the head-associated lymphoid tissues were identified using in situ immunohistochemical staining. Monoclonal antibodies specific for various lymphocyte surface antigens were used. The concentration of T-lymphocytes, particularly CD3+ and CD4+ cells, within the harderian gland increased with age, whereas the concentration of B cells remained the same. B-lymphocytes were observed within the germinal centers of the CALT of 4-week-old birds. The T lymphocytes within the CALT surrounded the B-cell-rich germinal centers. CD3+ T lymphocytes were the predominant cell population in all age groups examined. Increasing concentrations of CD3+, CD4+, and CD8+ T-lymphocytes were observed within the CALT as chicks developed from 1 week to 4 weeks of age. Finally, no changes were observed in lymphocyte populations within the CALT as chicks developed from 4 weeks to 8 weeks of age. PMID- 8719208 TI - Effect of serotype 2 and 3 Marek's disease vaccines on the development of avian leukosis virus-induced pre-neoplastic bursal follicles. AB - The effect of serotype 2 and 3 Marek's disease virus (MDV) vaccines on the development of pre-neoplastic bursal lesions induced by two strains of subgroup A avian leukosis virus (ALV) was studied. Chickens of line 15I5 x 7(1) susceptible to ALV-induced lymphoma were inoculated at hatch with Rous-associated virus-1 (RAV-1) or strain RPL-40 of subgroup A ALV. All chickens, except for negative controls, were also inoculated at hatch with strain 301B/1 of serotype 2 MDV or strain FC126 of turkey herpesvirus (HVT), a serotype 3 MDV. At 11 and 14 weeks of age, serial sections of bursal tissues from chickens in various treatment groups were stained with methyl green pyronin and examined for ALV-induced pre neoplastic bursal lesions, also known as hyperplastic follicles. At 6 days and at 11 and 14 weeks of age, bursal tissues from groups of chickens inoculated with serotype 2 MDV and ALV at hatch were also examined for the presence of MDV genome by in situ hybridization. The number of ALV-induced hyperplastic follicles was significantly higher in chickens inoculated with serotype 2 MDV than in unvaccinated chickens or in chickens vaccinated with HVT. In contrast, the lowest number of hyperplastic follicles was noted in chickens vaccinated with HVT, regardless of strain of ALV used. These results suggest that enhancement of lymphoid leukosis may result from an increase in the number of ALV-induced hyperplastic follicles caused by the MDV and also that the enhancing effect of MDV on ALV-induced lymphomagenesis may occur at the stage of formation of hyperplastic follicles in the bursa. PMID- 8719209 TI - Embryo vaccination of chickens with infectious bronchitis virus: histologic and ultrastructural lesion response and immunologic response to vaccination. AB - Chicken embryos 18 days of age and newly hatched chicks were vaccinated with an infectious bronchitis virus (IBV) vaccine (V-IBV) or with an IBV vaccine that had been serially passaged 40 times in chick kidney tissue culture (P-IBV). Immunologic and pathologic changes in the chicks were compared at selected intervals until the 35th day. Pathologic changes were evaluated by light, transmission, and scanning electron microscopy. Immunologic changes were assayed by a constant virus-diluting serum plaque-reduction test in chicken cell cultures, by 51Cr-release cytotoxicity assays, and by phytohemagglutination (PHA) responses. Embryos vaccinated with P-IBV and 1-day-old chicks vaccinated with V IBV had similar transient lesions that were confined primarily to the trachea. Embryo vaccination and posthatch vaccination induced similar primary and secondary antibody responses in chicks. It was concluded that neither vaccination technique consistently influenced PHA response of whole blood cells or natural killer cell reactivity of spleen effector cells. Additionally, effector cells cytotoxic to IBV-infected target cells were not detected in chicks vaccinated as embryos or at hatch. The pathologic and immunologic effects of vaccination with P IBV were comparable to those induced by conventional vaccination of chicks. PMID- 8719210 TI - Studies of intraspecies heterogeneity of mycoplasma synoviae, M. meleagridis, and M. iowae with arbitrarily primed polymerase chain reaction. AB - Intraspecies genotypic heterogeneity among isolates and strains of M. synoviae (MS), M. iowae (MI), and M. meleagridis (MM) was demonstrated with the arbitrarily primed polymerase chain reaction (AP-PCR) method. The generated banding patterns of amplified DNA indicated a degree of genotypic heterogeneity among MS, MI, and MM isolates and strains tested, whereas isolates from common sources generated clusters with identical banding patterns. The results show that flocks infected with members of the cluster were epidemiologically associated, possibly by a common or point source of infection. This AP-PCR method may be a useful identification tool in epidemiology of mycoplasmosis in poultry. PMID- 8719211 TI - Pathogenicity and persistence of Salmonella enteritidis and egg contamination in normal and infectious bursal disease virus-infected leghorn chicks. AB - The pathogenicity and persistence of Salmonella enteritidis (SE) phage type 8 and resulting egg contamination in normal and infectious bursal disease virus (IBDV) infected white leghorn chicks were evaluated over 34 weeks and in some birds over a 64-week period. Four hundred 1-day-old specific-pathogen-free (SPF) straight run white-leghorn chickens were allotted into four treatment groups: negative control, IBDV-infected, IBDV+SE-infected, and SE-infected. Chicks were infected with IBDV at 1 day of age and with SE phage type 8 at 2 days of age. SE persisted in the gut of more than 50% of the chicks of both SE-infected groups through 34 weeks postinoculation (PI), and SE could still be isolated from cloacal/rectal swabs taken at 64 weeks. IBDV+ SE-infected chicks had severe gross lesions and significantly (P < 0.001) higher mortality (32%) than the negative control (1%), IBDV-infected (10%), and SE-infected (1%) groups. Gross lesions consisting of fibrinous pericarditis, perihepatitis, peritonitis, airsacculitis, and inspissated yolk were observed only in the IBDV+SE-infected group. SE isolations from internal organs of chickens in the IBDV+SE-infected group decreased from 83% at 8 weeks to 0% at 14 weeks PI; isolations from the SE-infected group decreased from 50% at 8 weeks to 0% at 10 weeks PI. Salmonella isolations increased from 0% to 14% in both groups at 18 weeks, corresponding with the time of sexual maturity. Of the 1,050 eggs cultured from the IBDV+SE-infected group, SE was isolated from 88 shells, five albumens, and two yolks. In contrast, of 1,258 eggs from the SE-infected group, 33 shells and none of the albumens and yolks were positive for SE. All eggs that had SE-positive contents also had SE-positive shells. PMID- 8719212 TI - The identification of an 18,000-molecular-weight antigen specific to big liver and spleen disease. AB - Big liver and spleen (BLS) disease is an infectious syndrome of broiler breeders that has been serologically diagnosed worldwide with the agar gel immunodiffusion test. Liver homogenate from an affected broiler breeder was used as the antigen source in this study. This paper reports the identification, from liver, of a soluble basic protein antigen (molecular weight 18,000) that is specific to BLS disease. The antigen was partially purified from soluble extract of liver using a two-step fractionation procedure comprising Sephacryl S200 gel filtration and carboxymethyl (CM) cellulose cation exchange. After cation exchange, the partially purified (CM) antigen contained approximately 12 proteins. Immunoblotting was used to identify the single BLS disease-specific antigen. In addition, a polyclonal rabbit antiserum raised to the CM antigen was found to be monospecific to the 18,000-molecular-weight antigen by immunoblotting on the CM antigen. This serum was also of use in specifically detecting intracellular BLS disease antigen in frozen cryostat sections by indirect immunofluorescence. PMID- 8719213 TI - Quantification of experimental Salmonella enteritidis carrier state in B13 leghorn chicks. AB - Quantification of the carrier state of Salmonella enteritidis in chicks (i.e., persistent asymptomatic association of S. enteritidis with the host), should provide an optimized means for further investigations into this problem. We therefore developed an experimental carrier state model by oral inoculation of low doses (10(2)-10(4)) of S. enteritidis in B13 chicks at different ages. Liver, spleen, and ceca colonizations by the challenge strains were measured weekly by enumeration of S. enteritidis colony-forming units (CFU) for 7-12 weeks. High mortality rates, incompatible with the carrier state, were observed in chicks inoculated with 10(2) organisms of either a parental strain of S. enteritidis (5556) or a mutant resistant to streptomycin (Smr) and nalidixic acid (Nalr) (strain 1009) at 1 day old. Both strains colonized organs similarly, allowing us to use subsequently the SmrNalr mutant strain. The selected low doses of S. enteritidis induced no deaths in chicks inoculated at 1 or 3 weeks of age. However, inoculation of 3-week-old chicks did not induce a satisfactory carrier state; organ colonization by S. enteritidis was weak and transient, even after inoculation of 10(8) SE. In contrast, some birds infected at 1 week of age presented the challenge strain in the liver and spleen for 3 weeks after inoculation and in the ceca for 12 weeks postchallenge. Most of these birds were colonized by S. enteritidis in the liver and in the ceca for 3 weeks and 10 weeks, respectively, following inoculation. Generally, CFU levels were highest during the first week(s) after inoculation and then decreased progressively. Levels of S. enteritidis were lower in the liver and spleen than in the ceca. Oral inoculation of 1-week-old birds with 5 x 10(4) S. enteritidis provided the required model, allowing quantification of the carrier state of S. enteritidis in chicks. PMID- 8719214 TI - Avian mycoplasma identification using polymerase chain reaction amplicon and restriction fragment length polymorphism analysis. AB - A general mycoplasma polymerase chain reaction (PCR) was used to generate amplicon (DNA amplification product) from nine avian mycoplasma species. The PCR amplicons were reacted with 24 restriction enzymes, and the electrophoretic patterns of restriction fragment length polymorphism (RFLP) were evaluated for differences between species of mycoplasms. Four (DraI, MseI, RsaI, Tsp5091) of the 24 restriction enzymes cut the PCR amplicon of all nine mycoplasma species. The nine avian mycoplasma species could be distinctly differentiated using the RFLP analysis of the PCR amplicon. PMID- 8719215 TI - A genetic hybrid of the Campylobacter jejuni flaA gene with LT-B of Escherichia coli and assessment of the efficacy of the hybrid protein as an oral chicken vaccine. AB - The objectives of this study were to produce Campylobacter jejuni flagellin fused to the B-subunit of the labile toxin (LT-B) of Escherichia coli and to assess the efficacy of the hybrid protein as a chicken vaccine. Part of the flaA gene (780 base pairs) was cloned in plasmid pBEB downstream and in frame with the LT-B to allow expression of a hybrid protein. Transformed E. coli chi 6097 expressed the hybrid protein (43 kdaltons) in inclusion bodies at mid log phase. The inclusion bodies were isolated, and the identity of the protein was verified by western blot. This hybrid protein was administered as a vaccine to chickens either orally (0, 250, 500, or 1000 micrograms total protein) or intramuscularly (250 or 1000 micrograms). Alimentary secretions were collected, and specific antibodies were assayed by western blot analyses. Seventy-two percent of the birds vaccinated orally with 1000 micrograms protein showed detectable antibodies against C. jejuni flagellin in the excreta. None of the control birds produced detectable antibody to this antigen. For trials to demonstrate clearance of Campylobacter, groups of chickens were vaccinated with the hybrid protein at 2 and 4 wk of age and challenged at 3 wk with an excess of C.jejuni. The number of birds that remained colonized at 5 wk of age was significantly lower among the vaccinated birds than among controls. PMID- 8719216 TI - Experimental reduction of eggshell conductance during incubation. I. effect on the susceptibility to ascites syndrome. AB - Two hundred-four fertile broiler chicken eggs were obtained from a commercial source and divided into three equal groups. On day 1 of incubation, 68 eggs were selected randomly and four strips of vinyl tape were applied to the shell below the air cell (tape-day 1) to reduce eggshell conductance. This procedure was repeated with an additional group of 68 eggs on day 14 (tape-day 14). Sixty-eight eggs were incubated without treatment, as controls. One week after hatch, 20 chickens from each treatment group and control group were placed into a hypobaric chamber (simulated altitude of 2500 m) for 5 weeks. The remaining chickens in each group were maintained under normobaric conditions. The hematocrit and the mean frontal resultant electrical vector (MRV) of the heart were measured following 1 week, 3 weeks, and 5 weeks of hypobaric or normobaric exposure. Surviving chickens were euthanized at the end of 5 weeks. The weight ratio of right ventricle to left ventricle plus septum (the hypertrophy index [HI]) and the cardiac index, the HI divided by body weight, were determined. All mortality during the study was subjectively scored for the presence of ascites syndrome lesions. The percentage of chickens dying during, or exhibiting ascites syndrome at the completion of, the 5-week hypobaric exposure was 16.7%, 66.7%, and 58% for control, tape-day 1, and tape-day 14 treatments, respectively. MRV values of birds following hypobaric exposure were significantly different between treatment and control groups of the hypobaric exposure and between the two tape treatments. These results suggest that reducing conductance of the eggshell during incubation significantly potentiates the development of ascites syndrome in the broiler chicken. PMID- 8719217 TI - Evaluation of enrofloxacin against egg transmission of Mycoplasma gallisepticum. AB - Five groups of 20 commercial leghorn hens near peak production were challenged with Mycoplasma gallisepticum, and medicated with enrofloxacin in the drinking water at 7-11, 21-25, or 7-11 and 21-25 days postchallenge (PC), a combination of lincomycin/spectinomycin at 7-11 and 21-25 days PC, or left as untreated controls. Egg production records were maintained, and all eggs produced during the 63 days following challenge were incubated for 18 days and then cultured for M. gallisepticum. All groups experienced marked egg production drops beginning about 1 week PC; production returned to near normal levels after approximately 3 weeks. Under the conditions of this study water medication did not prevent egg production losses, probably because of the delay in the onset of medication, but medication may have resulted in a more rapid recovery of egg production. Enrofloxacin was highly effective in reducing the level of egg transmission of M. gallisepticum, although a small number of eggs (0-0.3%) in the treated groups were culture positive after medication was begun. Only 1 of 2463 live embryos was culture positive after the onset of treatment. In the unmedicated control group, 12.8% of all eggs were culture positive. Lincomycin/spectinomycin also reduced egg transmission (3.4% of eggs were culture positive after treatment was initiated), but not as effectively as enrofloxacin. Most of the egg transmission occurred before the medication could have had an effect. Both enrofloxacin and lincomycin/spectinomycin reduced the number of dead-in-shell embryos. PMID- 8719218 TI - Ascaridia dissimilis larval migration associated with enteritis and low market weights in meat turkeys. AB - A meat turkey ranch had a history of producing flocks with low market weights. Live bird samples and environmental samples from the ranch were collected over a period of eight months and analyzed at the California Veterinary Diagnostic Laboratory System-Turlock Branch. The disease condition consistently present was enteritis associated with Ascaridia dissimilis infection in birds from growout houses. After aggressive control measures for A. dissimilis were implemented, subsequent flocks reached expected market weights. PMID- 8719219 TI - Comparison of virus replication efficiency in lymphoid tissues among three infectious bursal disease virus strains. AB - In order to study replication efficiency of infectious bursal disease virus (IBDV) in lymphoid tissues, both the virus titers and the virus antigen titers in four lymphoid tissues were compared among chickens inoculated with Ehime/91 (about 50% mortality), J1 (no mortality), or K (attenuated) IBDV strains during 1 7 days postinoculation (PI). IBDV antigens in tissue homogenates were detected by an enzyme-linked immunosorbent assay. In the bursa of Fabricius, higher virus titers were maintained for 1-3 days PI in chickens inoculated with Ehime/91 or J1 strains, whereas the virus titers increased gradually and reached to the peak on 3 days PI in chickens inoculated with the K strain. There were no clear differences in both the virus and the virus antigen titers in bursae and thymus between chickens inoculated with Ehime/91 and J1 stains. However, the virus and/or the virus antigen titers in spleen and bone marrow of chickens inoculated with Ehime/91 strain were higher than those of the J1-inoculated chickens. Immunohistochemical analyses indicated that larger numbers of IBDV antigen positive cells were detected in spleen and bone marrow of the Ehime/91 group than in those of the J1 group. There was almost no detectable virus and virus antigens in thymus, spleen, and bone marrow of the K-inoculated chickens throughout the experiment. These results suggest that efficient replication of IBDV not only in the bursa but also in the spleen and the bone marrow may be required for clinical infectious bursal disease. PMID- 8719220 TI - Intratracheal infection of chickens with Salmonella enteritidis and the effect of feed and water deprivation. AB - The tissue distribution of Salmonella enteritidis in intratracheally inoculated chickens and the effect of deprivation of food and water on tissue distributions of the bacteria have been investigated. Seven-week-old specific-pathogen-free chickens were inoculated intratracheally with 10(2), 10(5), or 10(8) cells and orally with 10(5) cells. The intratracheally inoculated organisms entered the blood stream immediately after inoculation and produced generalized infection. Infection by the intratracheal route resulted in colonization of S. enteritidis in the cecum that was similar to infection by the oral route. The tissue distribution of S. enteritidis was markedly affected when chickens were deprived of food and water for a short time, demonstrating an increased susceptibility of chickens to S. enteritidis infection. This suggests that stresses such as food and water deprivation are one of of the causes of the rapid dissemination of S. enteritidis among chickens in poultry houses. PMID- 8719221 TI - In vivo role of tumor necrosis-like factor in Eimeria tenella infection. AB - The effect of tumor necrosis-like factor (TNLF) on the pathogenesis of coccidiosis was investigated. Injection of crude chicken TNLF enhanced the weight loss caused by Eimeria tenella infection. Rabbit polyclonal antibody against recombinant human tumor necrosis factor-alpha (rhTNF) partially restored E. tenella-induced weight loss in SC chickens, but not in TK chickens. However, injection of chickens with chicken TNLF, rhTNF, and rabbit serum against rhTNF had no significant effect on cecal lesions. Both SC and TK chickens produced circulating TNLF following primary, but not secondary infection, and SC chickens showed higher level of TNLF production than TK chickens. Peripheral blood leukocyte-derived macrophages from SC and TK chickens produced a significant amount of TNLF compared to the preinfection condition when cocultured with sporozoites. In general, macrophages from SC chickens produced higher levels of TNLF than those from TK chickens. No significant difference was observed between primary and secondary infection. These results suggest that the excessive TNF production may be involved in weight loss caused by E. tenella infection in SC chickens. PMID- 8719222 TI - Invasion of Caco-2 cells by Salmonella typhimurium (Copenhagen) isolates from healthy and sick chickens. AB - In a previous study, Salmonella isolates of sick birds were distinguished from those of apparently healthy birds by their high degree of invasion of tissue culture cells. In this study, a single pair of Salmonella isolates was examined to determine the source of this observed difference in invasion. When isolates were allowed to invade Caco-2 cells for 8 hours, the isolate from the sick bird (S) appeared to invade in greater numbers than did the isolate from the healthy bird (H). However, when invasion was distinguished from intracellular growth/survival, it was found that H invaded in greater numbers than S, but once inside the cell, H declined in number, and S increased. Inhibition of RNA, protein, and DNA syntheses lessened the degree to which both invaded. The presence of mannose inhibited invasion by S but did not appear to inhibit invasion by H. Trypsin treatment of monolayers affected invasion of S and H, whereas neuraminidase treatment did not. There was no significant difference noted between S and H in ability to adhere to fixed monolayers. Therefore, the two isolates tested differ in their mechanisms of entry into Caco-2 cells, the efficiency with which they invade, and their ability to survive within Caco-2 cells. PMID- 8719223 TI - Identification of Salmonella enteritidis from experimentally infected hens using a colorimetric DNA hybridization method. AB - Identification of Salmonella enteritidis from cloacally challenged commercial laying hens was studied by comparing bacterial isolations using conventional methods with detection by the use of the GENE-TRAK colorimetric DNA probe assay. More positive test results were obtained using the latter on days 14, 28, and 42 postchallenge, but the difference between the two methods was not statistically significant. Over the duration of the experiment, positive cloacal samples were statistically more frequent from a commercial strain of white leghorn hens when compared with a commercial brown egg-producing strain (28/60 vs. 9/57; chi-square 1 df = 12.9, P < 0.001). Eggs having various shell defects were produced by the infected hens only after Salmonella challenge. These defects included, in order of frequency, elongated shape, thin shells, off-white color (tints), small size, wrinkles, and pimples. No Salmonella could be recovered from 193 defective eggs, nor were positive isolates made from additional tests performed on 50 normal eggs. Proteus sp. was isolated from 10 eggs, however. Our observations demonstrate that the GENE-TRAK colorimetric method is comparable with conventional bacteriology for the identification of Salmonella in cloacal samples taken from laying hens. Moreover, the two methods demonstrate the existence of breed differences in susceptibility to S. enteritidis challenge. PMID- 8719224 TI - Effects of Fusarium moniliforme culture material containing known levels of fumonisin B1 in ducklings. AB - Fusarium moniliforme culture material containing fumonisin B1 (FB1) was fed to white Pekin ducklings from 1 to 21 days of age. Four dietary treatments were prepared with 0, 100, 200, and 400 mg FB1/kg ration. Ducklings fed rations containing FB1 had a dose-dependent decrease in feed intake and weight gain. Increasing levels of FB1 in the ration were associated with increasing absolute organ weights of liver, heart, kidney, pancreas, and proventriculus. Liver sphinganine to sphingosine ratios increased significantly in ducklings fed FB1. Two of eight ducklings fed a ration containing 400 mg FB1/kg died prior to the termination of the experiment. Mild to moderate hepatocellular hyperplasia was evident in all ducklings fed FB1. Mild to moderate biliary hyperplasia was also noted in the liver sections of ducklings fed 400 mg FB1/kg in the ration. Ducklings, like other poultry, are relatively resistant to the toxic effects of FB1. PMID- 8719225 TI - A sequential histopathologic and immunocytochemical study of chickens, turkey poults, and broiler breeders experimentally infected with turkey rhinotracheitis virus. AB - The histopathologic changes and the distribution of turkey rhinotracheitis virus (TRTV) antigen in the respiratory and reproductive tracts of experimentally infected chickens, turkey poults, and broiler breeders is described. TRTV antigen was detected using both immunofluorescent staining of cryostat sections and immunoperoxidase staining of formalin-fixed tissues. Viral antigen was observed associated with the cilia of the epithelial cells of turbinates, trachea, and lung. No TRTV antigen and no histopathologic changes were detected in the conjunctiva of any of the sacrificed birds, or in the reproductive tract and central nervous system of broiler breeders. The main histopathologic lesions and sites of TRTV replication were observed in the ciliated epithelial cells of turbinates and lung. These findings bring forward new information about pathologic changes and TRTV antigen distribution in tissues of experimentally infected birds. PMID- 8719226 TI - Production of avian polyomavirus seronegative budgerigars (Melopsittacus undulatus) from seropositive adults. AB - Adult budgerigars (Melopsittacus undulatus) with 2 years of breeding experience were removed from an aviary with enzootic avian polyomavirus (APV) disease and maintained in an isolation unit. Following a 7-month respite from breeding, these birds were allowed to breed without interruption for 2 years. Although the adults were seropositive both at the beginning and end of the experiment, all 102 of their offspring were seronegative. These data suggest that APV can be eliminated from a budgerigar aviary with the use of simple management techniques. PMID- 8719228 TI - An atypical fowl pox outbreak in broilers in southern Brazil. AB - An unusual fowl pox outbreak occurred in two integrated broiler operations. The uncommon characteristic of this outbreak was that the pox lesions were manifested in the feathered parts of the body, mainly in the posterior dorsal area and external part of the thigh. Diagnosis was made by means of histopathology and virus isolation. Severe losses were incurred at the processing plant as a consequence of condemnation due to dermatitis. Vaccination of day-old chicks with mild fowl pox vaccine combined with Marek's disease (HVT) vaccine in the area of the outbreak appeared to be important in controlling the disease. PMID- 8719227 TI - Efficacy of miconazole nitrate against favus in oriental breed chickens. AB - Miconazole nitrate 2% was tested for its efficacy against Microsporum gallinae (the causative agent in favus) in a flock of various Oriental breed (Shamo and Aseel) and crossbreed chickens. Six adult males showing clinical signs of favus were randomized into control and experimental groups. The males were maintained on individual tiecords on the range, with no physical contact between birds. The experimental birds had the affected areas washed with soap and water and dried, and an ointment of miconazole nitrate 2% was applied. The experimental birds received the treatment twice a day for 34 days. Scrapings from the affected areas of all birds were cultured at the beginning and end of the test. At the end of the treatment period, the control birds were still positive for M. gallinae, but the organism could not be cultured from the treated birds. PMID- 8719229 TI - Debilitating cutaneous poxvirus lesions on two captive houbara bustards (Chlamydotis undulata). AB - Poxvirus was isolated from cutaneous nodules on two young Houbara bustards (Chlamydotis undulata) bred in captivity in Saudi Arabia. Birds were emaciated and presented nodules on tibiotarso-tarsometatarsal joints, toes, and the carpal joint. Diagnosis was confirmed by histopathology, virus isolation on inoculated chorioallantoic membranes of embryonated chicken's eggs, and electron microscopy. Progressive leg lesions were extensive and interfered with walking, significantly debilitating the birds. Successful excisions of these lesions were performed. PMID- 8719230 TI - Natural infection with attaching and effacing Escherichia coli (O 103:H-) in chicks. AB - The existence of natural infection with attaching and effacing Escherichia coli (AEEC) in chicks was reported. Numerous Gram-negative bacilli were attached to the enterocyte in association with lesions characterized on a wavy appearance of the mucosal surface in the intestines of six chicks. Immunohistochemically, these bacteria reacted positively with antiserum to Escherichia coli (O 103). By electron microscopy, numerous colibacilli were seen to be closely attached to the surface membranes of the enterocytes. In regions of bacterial attachment, almost all cell microvilli were effaced, and some of the remaining ones were elongated and/or disoriented. Part of the cell membrane formed a cup invagination and pedestal-like protrusion associated with the attached bacteria. A concentration of electron-dense material was seen beneath the adherent organisms. Bacteriologically, numerous E. coli (O 103:H-) were isolated from the jejunal contents of two chicks and the E. coli did not produce verotoxin or enterotoxin. The characteristic lesions could also be induced in the cecal mucosa of young chicks experimentally inoculated with the isolated E. coli. This is the first report of natural infection in chicks with AEEC. PMID- 8719231 TI - Esophageal trichomoniasis in chickens. AB - Esophageal trichomoniasis has been rarely reported in chickens. At the California Veterinary Diagnostic Laboratory System-Turlock Branch, this disease was recently diagnosed in two cases submitted from backyard chicken flocks. The esophageal lesions observed were similar to those seen in several other important diseases of chickens. The causative trichomonad organisms were readily demonstrated on wet smears and by histologic studies. In both cases, the investigated flocks were afflicted with several concurrent diseases. California has experienced an increase in the number of small nontraditional chicken production operations. These facilities are sometimes in close proximity to commercial poultry operations and biosecurity barriers occasionally fail. The poor husbandry practices often used in these small flocks make them a potential reservoir for rare diseases such as trichomoniasis and also for disease organisms that are devastating to commercial poultry. PMID- 8719232 TI - Dual infection of chickens with pox and infectious laryngotracheitis (ILT) confirmed with specific pox and ILT DNA dot-blot hybridization assays. AB - Dual infection with fowl pox (FP) and infectious laryngotracheitis (ILT) was diagnosed as the cause of acute mortality in a flock of three age groups of Hy Line leghorn layers. The affected chickens had not been previously vaccinated against either FP or ILT. The diagnosis was confirmed by virus isolation, histopathology, and the use of specific pox and ILT genomic DNA probes in a dot blot hybridization assay. FP and ILT vaccinations were recommended to control mortality. The use of FP- and ILT-specific DNA dot-blot hybridization may be used as a routine diagnostic tool to differentiate between the two diseases, especially in atypical cases of either infection or to confirm the existence of the two diseases as a mixed infection in a flock of chickens. PMID- 8719233 TI - Prediction of sleep disorders induced by beta-adrenergic receptor blocking agents based on receptor occupancy. AB - beta-adrenergic receptor blocking agents (beta-blocking agents) have been widely used clinically for the treatment of various cardiovascular conditions. However, beta-blocking agents are liable to cause sleep disturbance, such as vivid dreams, nightmares, increased waking, and insomnia. The mechanisms of the sleep disorders are not known, but several may conceivably be responsible for these CNS-related side effects. In the present study, we hypothesized that the sleep disorders are induced by the blockade of central or peripheral beta 2 receptors and/or central serotonin (5-HT) receptors. To verify the hypothesis, we retrospectively analyzed the relationships between the extent of the sleep disorders and the beta 1, beta 2, or 5-HT receptor occupancies for four beta-blocking agents (atenolol, metoprolol, pindolol, and propranolol). No significant correlations were observed among pharmacokinetic/physicochemical parameters (therapeutic dose, plasma concentration, plasma unbound concentration, cerebrospinal fluid concentration, and lipid solubility) and pharmacodynamic parameters (the scores of the sleep disorders such as the number of dreams). Furthermore, no significant relationship (correlation coefficient: r < 0.3) was observed between beta 1 receptor occupancies of the drugs and the number of dreams. On the other hand, good relationships (r > 0.95) were observed between central and peripheral beta 2 or central 5-HT receptor occupancies and the number of dreams. These findings suggest that beta 2 and/or 5-HT receptor occupancy is superior to beta 1 receptor occupancy as an index for the sleep disorders. PMID- 8719234 TI - Influence of the rate of intravenous administration of eliprodil (SL 82.0715), a new anti-ischaemic agent, on its distribution in rat plasma and tissues. AB - This investigation studied the possible effect of different iv administration rates (bolus and infusions) of eliprodil, a new anti-ischemic agent, on the drug distribution in various body compartments. Following bolus administration of a 15 mg kg-1 dose, plasma concentrations were best fitted by a 3-compartment open model of t1/2 alpha = 14 sec, t1/2 beta = 4 min, and t1/2 gamma = 1.8 hr. Plasma and heart Cmax values were lowered by decreasing the infusion rate (the 15-mg dose was administered in 15 or 60 min) whereas brain Cmax values were not modified. In contrast, AUC values did not depend upon the rate of infusion. The present findings may have important implications in relating tissue concentrations with the desired therapeutic effect as well as with the side effects of the drug at its sites of action within brain and heart. The use of a simplified model built with plasma and tissue kinetic parameters following bolus injection allows one to predict the amount of drug present in the organs according to the mode of administration, but not the evolution of tissue to plasma ratio during the infusions. PMID- 8719235 TI - Third-generation model for corticosteroid pharmacodynamics: roles of glucocorticoid receptor mRNA and tyrosine aminotransferase mRNA in rat liver. AB - A third-generation pharmacokinetic/pharmacodynamic model was proposed for receptor/gene-mediated corticosteroid effects. The roles of the messenger RNA (mRNA) for the glucocorticoid receptor (GR) in hepatic GR down-regulation and the mRNA for hepatic tyrosine aminotransferase (TAT) induction by methylprednisolone (MPL) were examined. Male adrenalectomized Wistar rats received 50 mg/kg MPL iv. Blood and liver samples were collected at various time points for a period of 18 hr. Plasma concentrations of MPL, free hepatic cytosolic GR densities, GR mRNA, TAT mRNA, and TAT activities in liver were determined. Plasma MPL profile was biexponential with a terminal t1/2 of 0.57 hr. Free hepatic GR density rapidly disappeared from cytoplasm after the MPL dose and then slowly returned to about 60% of starting level after 16 hr. Meanwhile, GR mRNA level fell to 45% of baseline within 2 hr postdosing, and remained at that level for at least 18 hr. The GR down-regulation of GR mRNA and protein turnover rate were modeled. The TAT mRNA began to increase at about 2 hr, reached a maximum at about 5 hr, and declined to baseline by 14 hr. TAT induction followed a similar pattern, except the induction was delayed about 0.5 hr. Pharmacodynamic parameters were obtained by fitting seven differential equations in a piecewise fashion. The cascade of corticosteroid steps were modeled by a series of inductions for steroid-receptor DNA complex, two intermediate transit compartments, TAT mRNA, and TAT activity. Results indicate that GR mRNA and TAT mRNA are major controlling factors for the receptor/gene-mediated effects of corticosteroids. PMID- 8719236 TI - New hepatocellular diffusion model for analysis of hepatobiliary transport processes of drugs. AB - A new hepatocellular diffusion model was developed to kinetically evaluate the hepatobiliary transport processes of drugs in the perfusion system, based on the physiological structure of the liver. Since the equations describing the hepatocellular diffusion phenomena were derived as image forms in the Laplace domain, the fast inverse Laplace transform (FILT) was adopted to manipulate the image equations. Cefixime and cefpiramide were selected as model drugs. The concentrations in the perfusate and the excreted amounts into the bile were simultaneously measured at appropriate intervals after the rapid administration of each drug into the portal vein. The hepatocellular diffusion model was fitted to the biliary excretion profiles from rat livers, by means of a nonlinear least squares program, MULTI(FILT). According to this model, the hepatobiliary transport process of drug is kinetically separated into three steps, that is, the diffusion into and through the hepatocytes, the transfer from the hepatocytes into the bile canaliculi, and the movement through the bile canaliculi to the outlet of bile duct. These steps are characterized by the diffusion rate constant through hepatocytes (kdif), the permeability rate constant into the bile canaliculi (kbmc) and the transit time through the bile canaliculi to the outlet of bile duct (tcan), respectively. It was demonstrated that kdif of cefixime (0.023 min-1) was significantly smaller than that of cefpiramide (0.044 min-1), while the differences in kbmc and tcan were not obvious between cefixime and cefpiramide. kbmc and tcan of both drugs were about 1.2 min-1 and about 1.0 min, respectively. These parameters were correlated to the excretion ratio into the bile (Fbile) and the mean transit time from the sinusoid through the hepatocytes to the outlet of bile duct (tbile). PMID- 8719237 TI - Simulation for population analysis of Michaelis-Menten elimination kinetics. AB - A simulation study was conducted to compare the cost and performance of various models for population analysis of the steady state pharmacokinetic data arising from a one-compartment model with Michaelis-Menten elimination. The usual Michaelis-Menten model (MM) and its variants provide no estimate of the volume of distribution, and generally give poor estimates of the maximal elimination rate and the Michaelis-Menten constant. The exact solution to the Michaelis-Menten differential equation (TRUE) requires a precise analysis method designed for estimation of population pharmacokinetic parameters (the first-order conditional estimation method) and also considerable computational time to estimate population mean parameters accurately. The one-compartment model with dose dependent clearance (DDCL), in conjunction with the first-order conditional estimation or Laplacian method, ran approximately 20-fold faster than TRUE and gave accurate population mean parameters for a drug having a long biological half life relative to the dosing interval. These findings suggest that the well-known MM and its variants should be used carefully for the analysis of blood concentrations of a drug with Michaelis-Menten elimination kinetics, and that TRUE, in conjunction with a precise analysis method, should be considered for estimating population pharmacokinetic parameters. In addition, DDCL is a promising alternative to TRUE with respect to computation time, when the dosing interval is short relative to the biological half-life of a drug. PMID- 8719239 TI - Commentary on "Physiologically based pharmacokinetic modeling as a tool for drug development". PMID- 8719238 TI - Perspectives in pharmacokinetics. Physiologically based pharmacokinetic modeling as a tool for drug development. AB - Since the pioneering work of Haggard and Teorell in the first half of the 20th century, and of Bischoff and Dedrick in the late 1960s, physiologically based pharmacokinetic (PBPK) modeling has gone through cycles of general acceptance, and of healthy skepticism. Recently, however, the trend in the pharmaceuticals industry has been away from PBPK models. This is understandable when one considers the time and effort necessary to develop, test, and implement a typical PBPK model, and the fact that in the present-day environment for drug development, efficacy and safety must be demonstrated and drugs brought to market more rapidly. Although there are many modeling tools available to the pharmacokineticist today, many of which are preferable to PBPK modeling in most circumstances, there are several situations in which PBPK modeling provides distinct benefits that outweigh the drawbacks of increased time and effort for implementation. In this Commentary, we draw on our experience with this modeling technique in an industry setting to provide guidelines on when PBPK modeling techniques could be applied in an industrial setting to satisfy the needs of regulatory customers. We hope these guidelines will assist researchers in deciding when to apply PBPK modeling techniques. It is our contention that PBPK modeling should be viewed as one of many modeling tools for drug development. PMID- 8719240 TI - Transient expression of luciferase in Entamoeba histolytica driven by the ferredoxin gene 5' and 3' regions. AB - We have successfully transfected Entamoeba histolytica trophozoites with constructs containing ferredoxin sequences fused to the reporter gene luciferase. We have determined the conditions and parameters necessary to maximise transient luciferase expression in our system. Our optimal construct gave values of 268 x 10(3) relative light units per second (RLU s-1) when assayed representing a stimulation of 18,000-fold over the control vector. Comparison of differing constructs allowed us to conclude that the 5' and 3' ferredoxin sequences are both necessary for optimal luciferase expression from our vectors. Transcriptional initiation occurs within the consensus sequence ATTCA in both construct and chromosomal ferredoxin promoters. PMID- 8719241 TI - Plasmodium falciparum glutathione reductase exhibits sequence similarities with the human host enzyme in the core structure but differs at the ligand-binding sites. AB - The homodimeric flavoenzyme glutathione reductase (GR) which catalyzes the reduction of glutathione disulfide is a cornerstone of the malaria parasite antioxidant defense and repair mechanisms. Here we report on the identification of the GR gene from Plasmodium falciparum. A 1.4-kb fragment of the gene was amplified by polymerase chain reaction (PCR). Using this PCR fragment as a probe a full length cDNA clone (2085 bp) was isolated from a P. falciparum gametocyte library. The deduced amino acid sequence of 541 residues shows an overall identity of 35% when compared to the human enzyme. Most amino acids of known function are identical. However, notable differences between human and parasite protein occur in the glutathione-binding pocket (for instance, Glu374 instead of the expected basic residue) and at the intersubunit contact area. These regions are of particular interest since they represent binding sites of known GR inhibitors. Consequently, parasite GR can serve as a target structure for the design of antimalarial drugs. PMID- 8719242 TI - Characterisation of a novel Kunitz-type molecule from the trematode Fasciola hepatica. AB - A low molecular mass monomeric protein termed Fh-KTM (Fasciola hepatica Kunitz type molecule) was isolated from the trematode Fasciola hepatica. Fh-KTM is a single polypeptide of 58 amino acids and a Mr of 6751. The complete amino acid sequence of Fh-KTM was determined and revealed significant similarity to the Kunitz-type (BPTI) family of proteinase inhibitors. Several polymorphisms were observed suggesting that more than one Fh-KTM molecule may be expressed by this parasite. Modified proline residues were shown to occur at all four positions in this protein as 3-hydroxy derivatives. This is the first report of 3 hydroxyproline residues in a Kunitz-type molecule. Indirect immunofluorescence and immunogold labelling revealed that Fh-KTM is an abundant molecule within the parasite localised to the gut, the parenchymal tissue and the tegument of adult F. hepatica. Serine protease inhibition assays revealed that Fh-KTM exhibited little or no inhibition against chymotrypsin, kallikrein, urokinase or key serine proteases of the blood coagulation pathways. However, Fh-KTM was able to inhibit trypsin even though the P1 reactive amino acid of Fh-KTM was a leucine residue. PMID- 8719243 TI - A polymorphic minisatellite sequence in the subtelomeric regions of chromosomes I and V in Leishmania infantum. AB - A minisatellite DNA sequence is described for the first time in Leishmania infantum. It is borne by four chromosomes and consists of an 81-bp repeat unit organised in several clusters. On chromosomes I and V of L. infantum, the clusters are tightly located in the size-variable subtelomeric regions. The organisation of this sequence may be related to that of the subtelomeric interspersed repeat sequences identified in the human genome. The sequencing of seven repeat units, some subcloned from the same cluster, allowed the definition of a consensus sequence of 81 bp, particularly G/C rich (73%). Two subfamilies were clearly defined: one exhibits a 91-95% homology with the consensus sequence; the second one comprises two monomers sharing a 91% homology but only 77% homology with the consensus sequence. The two types of monomers can be found in the same cluster. These data suggest interactions between monomers and a possible role of this sequence in the instability of these regions. Finally, restriction fragment length polymorphisms were revealed by this sequence among various strains of L. infantum. Besides allowing the detection of recombination events in the unstable regions of the chromosomes, this new marker may become a useful tool in the study of the parasite population dynamics in leishmaniasis foci. PMID- 8719244 TI - The secretary pathway of plasmodium falciparum regulates transport of p82/RAP1 to the rhoptries. AB - The rhoptries of Plasmodium falciparum are formed during a restricted period in the asexual erythrocytic cycle. The steps required for rhoptry biogenesis and the pathway for targeting proteins to the rhoptries have not been elucidated. Using the maturation of the Rhoptry-Associated Protein 1 (RAP-1) gene product to study these steps, it is reported here that a secretory pathway controls transport of protein complexes containing RAP-1 products to the rhoptries. Both brefeldin A (BFA) and low temperature reversibly block the processing of an 86-kDa precursor (Pr86) to the mature 82-kDa RAP-1 product (p82). Furthermore, the points of action of BFA and low temperature appear to overlap since their sequential application reversibly prevents Pr86 processing. Treatment of intact cells with N ethylmaleimide, which prevents the fusion of transport vesicles with Golgi membranes in other eukaryotic cells, irreversibly blocks processing of Pr86. The role of the secretory pathway in targeting p82 protein complexes to the rhoptries product of RAP-1. These in vitro results also reveal that the RAP-1 product contains a cleavable N-terminal signal peptide and appears to be initially synthesized as an 84-kDa protein. The above data indicate that transport of p82 to the rhoptries is regulated by the secretory pathway and that the RAP-1 primary translation product differs in apparent molecular weight from the in vivo precursor Pr86. Our results suggest that rhoptry biogenesis is controlled in part by the secretory pathway and that the RAP-1 gene product acquires a previously undetected protein modification during its maturation. PMID- 8719245 TI - Restriction enzyme-mediated integration elevates transformation frequency and enables co-transfection of Toxoplasma gondii. AB - This report describes the use of restriction enzyme-mediated integration (REMI) to increase the transformation frequency and allow co-transfection of several unselected constructs under the selection of a single selectable marker. We found that while BamHI (the enzyme used to originally demonstrate REMI (Schiestl, R.H. and Petes, T.D. (1991) Integration of DNA fragments by illegitimate recombination in Saccharomyces cerevisiae. Proc. Nati. Acad. Sci. USA 88, 7585-7589) increased the number of transformants by 2-5-fold over the control without added enzyme, NotI proved to be a further 29-46-times more effective in enhancing stable transformation. This simple technique was used in the transformation of three non selective markers (two modified membrane proteins and beta-galactosidase) with a selectable construct expressing chloramphenicol acetyltransferase. Following chloramphenicol selection, four out of ten independent transformants stably acquired all four constructs with at least two expressing all four genes at the protein level. These results demonstrate that REMI may be used in the efficient stable transformation and co-transfection of this and perhaps other protozoan parasites. PMID- 8719246 TI - Glycosyl inositolphospholipid-anchored structures in Herpetomonas davidi. AB - Glycosyl inositolphospholipid (GPI)-anchored structures in the monogenetic parasite Herpetomanas davidi, were labeled with [3H]glucosamine, and characterized by enzymatic and chemical treatments that are typical for the identification of GPI anchors. [3H]Myristate incorporated into two different pools of GPI-linked structures that could be separated by chromatography on octyl Sepharose. One pool consisted of three GPI-anchored proteins with apparent molecular masses of 21,31 and 45 kDa, and the GPI lipid moieties were identified as alkyl-lysoglycerols. The label in the other pool associated with lipopeptidophosphoglycan (LPPG)-like structures of approximately 12-kDa molecular mass, containing ceramide-type GPI lipid anchors. While protein GPI anchors could also be labeled using [3H]glucosamine as radiolabeled GPI anchor precursor, hardly any radioactivity was incorporated into the LPPG-like structures. H. davidi is one of the few organisms identified to date that synthesizes two structurally different lipid moieties for GPI anchoring of membrane components. PMID- 8719247 TI - Glucose transport in amastigotes and promastigotes of Leishmania mexicana mexicana. AB - Promastigotes and amastigotes of Leishmania mexicana mexicana transported 2-deoxy D-glucose (2-DOG) by a saturable process with a Km of 24 +/- 3 microM and Vmax of 2.21 nmol min-1 (mg protein)-1 for the promastigote and a Km of 29 +/- 8 microM and Vmax of 0.13 nmol min-1 (mg protein)-1 for the amastigote stage. Amastigotes incorporated 2-DOG maximally at pH 5.0, while for promastigotes the optimum was at pH 7.0. Mid-log phase promastigotes were found to accumulate 2-DOG via a stereospecific carrier-mediated process which was competitively inhibited by D glucose and D-mannose but not L-glucose. Transport was dependent upon temperature, with a Q10 in promastigotes of 1.83 and an optimum rate at 35 degrees C (+/- 4 degrees C) with an activation energy of 50.12 kJ mol-1. Stationary phase promastigotes accumulated 2-DOG at approximately twice the rate of mid-log phase promastigotes. Cytochalasin B, forskolin and phloretin were all found to inhibit human erythrocyte 2-DOG uptake but only cytochalasin B was found significantly to inhibit promastigote 2-DOG uptake. Interestingly, leishmanial 2 DOG uptake was inhibited by a series of membrane potential antagonists including the ionophore monensin, the H+ATPase inhibitor N, N'-dicyclohexylcarbodiimide (DCCD) and uncoupling agent carbonylcyanide-4-(triflouromethoxy) phenylhydrazone (FCCP), as well as, the tricyclic drugs chlomipramine and imipramine, but was insensitive to the Na+/K+ATPase inhibitor ouabain and the antitrypanosomal drugs Pentostam and Suramin. We therefore conclude that there are significant structural and mechanistic differences between the D-glucose uptake systems of Leishmania and the mammalian host to merit the inclusion of glucose transporters as putative targets for rational drug design. PMID- 8719248 TI - Complementation of a Toxoplasma gondii ROP1 knock-out mutant using phleomycin selection. AB - The ROP1 gene of Toxoplasma gondii encodes a rhoptry protein that has been implicated in host cell invasion by this obligate intracellular protozoan. To further explore the function of this protein, we created a ROP1 deletion mutant by transfection with a plasmid encoding the bacterial chloramphenicol acetyltransferase (cat) gene flanked by ROP1 genomic sequences. Selection for chloramphenicol resistance yielded the desired ROP1-deleted or 'knock-out' mutant. Analysis of this mutant both in vitro and in vivo shows no significant alterations in growth rate, host specificity, invasiveness or virulence and thus the ROP1 gene is not obligatory for the RH strain, at least under the conditions tested. However, electron microscopy reveals that the mutant strain's rhoptries are altered in ultrastructure; they are thinner and homogeneously electron-dense compared with the thicker and normally mottled or honeycombed appearance of wild type rhoptries. The knock-out mutant was rescued using co-transfection of a cosmid carrying the ROP1 gene together with a plasmid encoding a new selectable marker for T. gondii, the bleomycin resistance gene (ble) from Streptoalloteichus. Southern blot analysis showed that both DNAs were stably integrated into the Toxoplasma genome, although not into the ROPI locus. The resulting strain showed wild-type levels of ROP1 expression and rescue of the ultrastructural phenotype (i.e., the rhoptries returned to their normal, mottled appearance), thus establishing a cause/effect relationship between the absence of ROP1 and the electron-opacity. These results demonstrate the utility of the reverse genetic approach in the study of Toxoplasma gene function and provide a further selectable marker for such manipulations. PMID- 8719249 TI - Cloning and expression of the adenine phosphoribosyltransferase gene from Leishmania donovani. PMID- 8719250 TI - The Plasmodium cynomolgi merozoite surface protein 1 C-terminal sequence and its homologies with other Plasmodium species. PMID- 8719251 TI - Human serum resistant Trypanosoma brucei rhodesiense accumulates similar amounts of fluorescently-labelled trypanolytic human HDL3 particles as human serum sensitive T.b. brucei. PMID- 8719252 TI - Molecular characterization of the mitochondrial heat shock protein 60 gene from Trypanosoma brucei. PMID- 8719253 TI - Merozoite surface antigen 2 (MSA-2) gene of Plasmodium falciparum strains from India. PMID- 8719254 TI - A re-evaluation of the determinants of glomerular filtration rate. AB - Several factors are potentially able to change the glomerular filtration rate (GFR) and thereby participate in its regulation, but only a few factors seem to be physiologically important. The variable nature of proximal tubular pressure should be recognized as important in the regulation of GFR. It is argued that a distinction should be made between the terms 'autoregulation of GFR' and 'regulation of GFR'. The tubuloglomerular feedback mechanism (TGF) is an important factor for autoregulatory control of GFR. When perturbations result in major increases in tubular flow, the TGF saturates. Proximal tubular pressure then increases and becomes the major factor responsible for the stabilization of GFR. Changes in the proximal reabsorption rate (APR) are important for long-term variations in GFR (regulation of GFR). Small changes in the APR cause near parallel changes in the GFR mainly through the TGF mechanism, while larger changes in the APR cause near parallel changes in the GFR mainly because of the effect on tubular pressure. The hydraulic resistance in the distal nephron segments is an additional factor in regulating GFR, through its effect on proximal tubular pressure. The stimulus to the TGF mechanism also depresses renin release. The resulting local angiotensin II concentration has effects both on the arteriolar resistances and on the APR. The renin-angiotensin system and TGF are therefore considered to be integrated parts of a common control system regulating GFR. According to the hypothesis advocated here, TGF-mediated changes in afferent arteriolar resistance and angiotensin-mediated changes in efferent arteriolar resistance and APR cooperate in counteracting perturbations in proximal tubular pressure and Henle loop flow. However, because of the biphasic proximal effect of angiotensin II, a major unresolved question is whether physiological increases in endogenous local angiotensin II concentrations stimulate or inhibit proximal reabsorption. PMID- 8719255 TI - Correlation between chloride flux via the mitochondria-rich cells and transepithelial water movement in isolated frog skin (Rana esculenta). AB - The coupling between net transepithelial Cl- influx and net water flow was investigated. Experiments were performed on isolated frog skin bathed in isotonic Cl- Ringer's solution in the presence of the Na+ channel blocking agent amiloride in the mucosal solution. The skins were voltage-clamped at -80 or -100 mV (with the serosal solution as reference). Under these conditions the current across the skin is carried by an influx of Cl-. In the absence of antidiuretic hormone the correlation between current and net water flow was low, but in the presence of the antidiuretic hormone, arginine vasotocin, there was a highly significant correlation between current and net water flow. The data presented here indicate that under steady state conditions about 70 molecules of water follow each Cl- ion across the skin. If the water influx is driven by electroosmosis one would expect that a change in current should result in an immediate change in the water flow. There was, however, a considerable time delay between the change in current and water flow. This indicates that the observed coupling between Cl- flux and water flow is caused by current-induced local osmosis and not electroosmosis. PMID- 8719256 TI - Ca selectivity of the transduction channels in the hair cells of the frog sacculus. AB - The extracellular receptor currents evoked by step displacements of the otolithic membrane of the isolated saccular macula of Rana esculenta were recorded under transepithelial voltage clamp conditions. With the aim to depolarize the hair cells and increase the fractional resistance of the apical membranes, the basal side of the preparation was bathed in saline with an increased K+ concentration (62 mM). This caused a shift in the non-linear receptor current-voltage relation along the voltage axis of -51 mV +/- 10 mV; (mean +/- SD; n = 32) and a reduction in the transepithelial resistance of 10%. Under these conditions the electrical properties of the macula are assumed to be controlled by the apical membranes. The effects of different concentrations of Ca2+ in the apical solution on the receptor current-voltage relation were examined. Change of the apical Ca2+ concentration (range 3 mM to 70 microM) varied the transepithelial voltage at which the receptor current was zero (Vrev). Fitting a modified constant field equation to the relation between the apical Ca2+ concentration and the change in Vrev gave an estimate of PCa/PK of the transduction channels of 212. Furthermore, a high relative permeability of the transduction channels for other divalent cations (Ba2+, Sr2+) was measured, whereas Mn2+ inhibited the receptor current. The receptor current was inhibited by amiloride (IC50 3.2 microM +/- 1.7 microM) and nifedipine (IC50 1.9 microM +/- 0.6 microM). Reduction of the apical Ca2+ concentration to 90 microM in standard apical solution reduced the size of the receptor current to 67% +/- 30% (n = 17) compared to control but did not affect the shape of the receptor current-voltage relation. Subsequent substitution of K+ by Na+ caused a further reduction of the receptor current to 32% +/- 29% (n = 9), changed the receptor current-voltage relation into a linear relation and diminished the adaptation of the receptor current. These results indicate that the mechano-electrical transduction channels of the frog saccular hair cells are highly selective to Ca2+ and that the conductance of the channels may be influenced by the apical monovalent cation species. PMID- 8719257 TI - Contraction specific changes in passive torque in human skeletal muscle. AB - The present investigation examined passive torque and electromyographic response in human skeletal muscle during passive static stretch within 60 s after maximal repetitive eccentric and concentric contractions. Passive torque (Nm) offered by the hamstring muscle group was measured during passive knee extension in a modified dynamometer in 10 subjects. The distal thigh was elevated to 0.52 rad from horizontal and the backrest was positioned at 1.57 rad. The lever arm moved the leg passively at 0.09 rad s-1 from a starting position of 1.48 rad below horizontal to the final position where it remained stationary for 90 s. Gross electrical activity of the human hamstring muscle group was recorded simultaneously. The effect of concentric or eccentric contraction was tested on separate days. Two stretch manoeuvres with a 45 min hiatus were administered on a control and experimental side. The experimental side performed 40 maximal effort repetitive concentric or eccentric hamstring muscle contractions at 1.05 rad s-1 prior to the second stretch. Passive torque during the 90 s stretch declined 30 35% on the experimental and control side in all stretches, P < 0.001, without a significant effect of prior contraction mode. Passive peak and final torques were lower on the experimental side, P < 0.01 after concentric contractions. Passive peak and final torques remained unchanged after eccentric contractions on the experimental side. The low level EMG response of the hamstring muscle during the stretch was unchanged after maximal repetitive concentric or eccentric contractions. These data demonstrate acute contraction specific alteration in passive torque in human skeletal muscle, which cannot be accounted for by EMG activity. Furthermore, the lack of difference on the control side implies that one 90 s stretch has no effect on passive torque of the muscle 45 min later. PMID- 8719258 TI - Effect of creatine supplementation on intramuscular TCr, metabolism and performance during intermittent, supramaximal exercise in humans. AB - This study examined the effect of (a) creatine supplementation on exercise metabolism and performance and (b) changes in intramuscular total creatine stores following a 5 day supplementation period and a 28 day wash-out period. Six men performed four exercise trials, each consisting of four 1 min cycling bouts, punctuated by 1 min of rest followed by a fifth bout to fatigue, all at a workload estimated to require 115 or 125% VO2,max. After three familiarization trials, one trial was conducted following a creatine monohydrate supplementation protocol (CREAT); the other after 28 d without creatine supplementation, in which the last 5 d involved placebo ingestion (CON). Intramuscular TCr was elevated (P < 0.05) in CREAT compared with the final familiarization trial (FAM 3) and CON. Concentrations of this metabolite in these latter trials were not different. In addition, a main effect (P < 0.05) for treatment was observed for PCr when the data from CREAT were compared with CON. In contrast, no differences were observed in the total adenine nucleotide pool (ATP+ADP+AMP), inosine 5'-monophosphate, ammonia, lactate or glycogen when comparing CREAT with CON. Despite the differences in TCr and PCr concentrations when comparing CREAT with other trials, no difference was observed in exercise duration in the fifth work bout. These data demonstrate that creatine supplementation results in an increase in TCr but this has no effect on performance during exercise of this nature, where the creatine kinase system is not the principal energy supplier. In addition 28 d without supplementation is a sufficient time to return intramuscular TCr stores to basal levels. PMID- 8719259 TI - Regional myosin heavy chain expression in volume and pressure overload induced cardiac hypertrophy. AB - Although the overall shift towards the V3 myosin heavy chain (MHC) has been shown to be associated with cardiac hypertrophy, quantitative evidence describing regional expression is sparse. The aim of this study was to compare and contrast the regional ventricular myosin isoform expression in two distinct haemodynamic states: pressure and volume overload. Volume overload was achieved using an aortocaval fistula (ACF) model and pressure overload by two-kidney-one-clip (2K1C) hypertension. A separate group (UC-2K1C) had the clip removed 1 week prior to investigation. Sham operated rats (SHAM) served as controls. All groups were studied 4 weeks after surgery. Ventricular tissue samples (approximately 50 mg) were taken from the walls of the right ventricle (RV), septum and left ventricular (LV) free wall. Tissue samples (excluding RV) were divided into endocardium and epicardium, and myosin expression was determined using polyacrylamide gel electrophoresis. Cardiac hypertrophy was substantial in both LV (1.7-fold) and RV (1.9-fold) in ACF rats. The 2K1C rats had similar LV enlargement (1.6-fold) whereas RV hypertrophy was not as great (1.2-fold). Blood pressure (BP) was increased 65% in 2K1C rats, whereas there was no change in ACF rats with respect to SHAM animals. After unclipping (UC-2K1C), LV hypertrophy and BP had returned towards control levels. In general, V3 MHC expression was associated with increasing LV hypertrophy in both 2K1C and ACF models. However, there was a marked endo-epi differential (1.5:1) in the LV free wall and septum of 2K1C rats. In contrast, in ACF rats there was no differential V3 MHC expression in the LV or septal tissue, i.e. expression was similar in both endo- and epi-samples. Elevated expression of V3 MHC persisted despite normotension and regression of cardiac hypertrophy in UC-2K1C rats. Taken together with published results demonstrating that relative transmural myocyte hypertrophy in ACF rats (endo > epi) is in contrast to that seen in 2K1C rats (epi > endo), the present findings reveal that regional V3 myosin expression represents a distinct adaptational component of the overall cardiac hypertrophic response in both volume and pressure overload. PMID- 8719260 TI - Endothelin-induced contractions in human placental blood vessels are enhanced in intrauterine growth retardation, and modulated by agents that regulate levels of intracellular calcium. AB - Endothelin-1 (ET-1) is a strongly vasoactive polypeptide that may be involved in the regulation of the uteroplacental blood flow. In the present study we have examined the contractile response to ET-1 in human placental arteries in the presence of several agents that interfere with storage of intracellular calcium, e.g. caffeine, ryanodine and thapsigargin. We have also compared the contractile response to ET-1 in normal pregnancies with that of patients with foetal intrauterine growth retardation (IUGR), a condition with reduced uteroplacental blood flow. We found that the response to ET-1 in the placental arteries from women with normal pregnancies was reduced by 20% in the absence of extracellular calcium. Caffeine relaxed the basal tone of the vessels and reduced the contractile response to ET-1 by 51%. Nifedipine in addition to caffeine resulted in a reduction of 70%. Ryanodine also reduced the tone. Thapsigargin had no effect on the placental arteries at lower concentrations, but gave a progressive and slow contraction at 10(-6) M. The ET-1 induced contraction in placental arteries from IUGR patients was 67% more potent than in placental arteries from women with normal pregnancies, 129% as compared with 77% of the maximal K(+) induced contraction. We conclude that the ET-1-induced contractile response in the human placental artery is dependent on influx of extracellular calcium as well as mobilization of calcium from intracellular stores. An increased sensitivity to ET-1 in placental arteries may contribute to the reduced uteroplacental blood flow in intrauterine growth retardation. PMID- 8719261 TI - The bovine central adrenomedullary vein: a target for endothelins. AB - This study reports on morphological and contractile properties of the bovine central adrenomedullary vein (bCAMV). Up to several layers of circularly orientated smooth muscle cells (SMC) were observed, however, without forming a continuous, closed sheath. Discrete bundles of eccentrically arranged, longitudinal SMC were also conspicuous. Chromaffin cells were in most cases located outside the SMC layers, while sometimes being in close apposition to the endothelium in areas without SMC. Circularly mounted preparations of the endothelium-denuded vessel responded selectively to high K+, endothelins (ETs) and neuropeptide Y (NPY). The threshold for ET-1 was 0.13 nM and the half maximally effective concentration (EC50) was 3 +/- 1 nM (n = 9). The order of potencies was ET-1 > or = ET-2 >> ET-3, suggesting a vascular receptor (ETA). Concentrations at and above EC50 frequently developed long-lasting oscillations during the spontaneous relaxation of the ET-1 evoked tension. This response was partly (21%) independent of extracellular Ca2+. A marked tachyphylaxis developed to ET-1 (3-30 nM), resulting, on the other hand, in facilitation of the subsequent constrictor responses to high K+ and NPY. Propranolol and phentolamine alone, or in combination, were without effects on the basal tension and on the above-mentioned responses to high K+, ET-1 or NPY, making a contribution from adrenoceptor activation unlikely. No response was obtained with exogenous catecholamines, acetylcholine or serotonin, nor with a series of peptides known to occur in the adrenal medulla. This study shows that bCAMV is not a passive capacitance vessel but appears unique among mammalian veins in being selectively regulated by ETs. PMID- 8719262 TI - Influence of forebrain lesions and isoflurane anaesthesia, respectively, on responses to the intracarotid infusion of angiotensin II in sheep. AB - In conscious sheep, lesions involving the anterior wall of the third cerebral ventricle extinguished the dipsogenic and cortisol releasing effects of 10 min intracarotid (i.c.) infusions of angiotensin II (Ang II) (20 pmol kg-1 min-1). The sustained hypertension obtained in response to i.c. Ang II in the intact animal was not significantly changed by the forebrain lesions. Pronounced, but short-lasting tachycardia immediately developed in response to i.c. Ang II in control animals. After a 1 min return to initial values the heart rate (HR) rose to about 20 beats min-1 above pre-infusion level during the remaining infusion period. After a brief post-infusion fall, HR retook that level. It then gradually declined but did not return to initial level until about 25 min later. During isoflurane anaesthesia in the latter animals carotid blood pressure (cBP) was reduced by approximately 30% while the HR was more than doubled. The cBP rise in response to i.c. Ang II infusion was < 50% of that seen in awake animals and the pressure returned to initial level within approximately 5 min after the end of the infusion. The infusion did not influence the HR. This study provides support for the idea that blood borne Ang II, bound at cerebral sites posterior to the hypothalamic region, continues to have cardiovascular effects for approximately 20 min after the concentration in the blood has returned to pre-infusion level. PMID- 8719263 TI - Sympathetic influence on cardiovascular responses to sustained head-up tilt in humans. AB - Sympathetic beta-adrenergic influences on cardiovascular responses to 50 degrees head-up tilt were evaluated with metoprolol (beta 1-blockade; 0.29 mg kg-1) and propranolol (beta 1 and beta 2-blockade; 0.28 mg kg-1) in eight males. A normotensive-tachycardic phase was followed by a hypotensive-bradycardic episode associated with presyncopal symptoms after 23 +/- 3 min (control, mean +/- SE). Head-up tilt made thoracic electrical impedance (3.0 +/- 1.0 omega), mean arterial pressure (MAP, 86 +/- 4-93 +/- 4 mmHg), heart rate (HR, 63 +/- 3-99 +/- 10 beats min-1) and total peripheral resistance (TPR, 15 +/- 1-28 +/- 4 mmHg min L-1) increase, while central venous oxygen saturation (74 +/- 2-58 +/- 4%), cardiac output (5.7 +/- 0.1-3.1 +/- 0.3 L min-1), stroke volume (95 +/- 6-41 +/- 5 mL) and pulse pressure (55 +/- 4-49 +/- 4 mmHg) decreased (P < 0.05). Central venous pressure decreased during head-up tilt (7 +/- 2-0 +/- 1 mmHg), but it remained stable during the sustained tilt. At the appearance of presyncopal symptoms MAP (49 +/- 3 mmHg), HR (66 +/- 4 beats min-1) and TPR (15 +/- 3 mmHg min L-1) decreased (P < 0.05). Neither metoprolol or propranolol changed tilt tolerance or cardiovascular variables, except for HR that remained at 57 +/- 2 (metoprolol) and 55 +/- 3 beats min-1 (propranolol), and MAP that remained at 87 +/- 5 mmHg during the first phase with metoprolol. In conclusion, sympathetic activation was crucial for the heart rate elevation during normotensive head-up tilt, but not for tilt tolerance or for the associated hypotension and bradycardia. PMID- 8719264 TI - Distribution and function of nitric oxide-containing nerves in canine corpus cavernosum and spongiosum. AB - Specimens of penile erectile tissue from the corpus cavernosum (CC) and corpus spongiosum (CS) of beagle dogs were investigated with reference to morphological and functional aspects of the nitric oxide (NO) system. Using immunohistochemistry, the smooth muscle bundles of the CC were found to contain a rich innervation, as seen by the pan-neuronal marker, protein gene product 9.5. A large number of nerves also stained positively for acetylcholine esterase. There was a moderate to rich supply of nerves containing NO synthase, which was frequently co-localized with vasoactive intestinal polypeptide. CS preparations from the proximal penis (the spongious bulb) had an innervation similar to that of the CC. However, CS preparations from the distal penis had a distinctly lower density of nerves than the proximal part, and spongious tissue from the glans penis was practically devoid of nerves. In isolated preparations of proximal and distal CC and CS, contracted by noradrenaline, electrical field stimulation (EFS) produced frequency-dependent relaxations. The responses were less pronounced in preparations from the distal penis, particularly in the CS. Pre-treatment with the NO-synthesis inhibitor NG-nitro-L-arginine abolished all relaxations. Carbachol effectively relaxed noradrenaline-contracted strips. The responses were similar in proximal preparations from the CC and CS, and significantly less pronounced in strips from the distal penis. NG-nitro-L-arginine significantly reduced the relaxant effect of carbachol. Preparations of spongious tissue from the glans penis did not react to high potassium solution, addition of noradrenaline, or EFS. The results demonstrate regional differences within the CS, both in innervation and functional responses. PMID- 8719265 TI - Hyperglycaemia is responsible for the inhibited gastrointestinal transit in the early diabetic rat. AB - The effect of plasma glucose levels on the gastrointestinal motility of the rat was studied. Chronic hyperglycaemia was induced by i.v. injection of streptozotocin 1 week before the motility experiment. Some rats received additional daily insulin therapy (1.25, 2.5 or 10 IU kg-1) after induction of diabetes mellitus. Acute hyperglycaemia was induced by the continuous i.v. infusion of glucose solution (11, 22, 44 or 88 mg kg-1 min-1) 10 min before the motility experiments. The rats were killed 15 min after successful orogastric feeding of a charcoal-contained suspension. Gastrointestinal transit was calculated as the percentage of the overall length of the small intestine to which the charcoal moved during this time period. The diabetic rats were found to have delayed transit compared with controls (mean +/- SEM: 32.2 +/- 2.1% vs. 42.9 +/- 4.2%, P < 0.05). Correction of hyperglycaemia with moderate doses of insulin therapy failed to inhibit transit, whereas hypoglycaemia induced by high-dose insulin treatment enhanced transit. High doses of glucose elicited acute hyperglycaemia and delayed transit when compared with saline infused non-diabetic rats. In early diabetes, hyperglycaemia probably mediates the inhibited gastrointestinal transit, since correction of hyperglycaemia usually restores the delayed transit. PMID- 8719266 TI - Retinal haemorrhage during sustained hypobaric hypoxia in rats. PMID- 8719267 TI - Extracellular dopamine in the nucleus accumbens of the rat during treadmill running. PMID- 8719268 TI - Nitric oxide, produced in the upper airways, may act in an 'aerocrine' fashion to enhance pulmonary oxygen uptake in humans. PMID- 8719269 TI - Cyclosporin A dramatically ameliorates CA1 hippocampal damage following transient forebrain ischaemia in the rat. PMID- 8719270 TI - Nitric oxide occurs in high concentrations in monkey upper airways. PMID- 8719271 TI - Histamine promotes leucocyte recruitment by prolonging chemoattractant-induced leucocyte adhesion in the rat mesentery. PMID- 8719272 TI - Allergen-induced late airways reactions in the pig. Influence of endogenous and exogenous glucocorticoids. PMID- 8719273 TI - Cajal's nucleus of the stria medullaris: characterization by in situ hybridization and immunohistochemistry for enkephalin. AB - While analyzing the distribution of enkephalinergic neurons by in situ hybridization and immunohistochemistry in the septal region of untreated or colchicine-injected rats, a densely packed enkephalinergic group of neurons was identified that corresponds to a small nucleus first described by Cajal as the nucleus of the stria medullaris. It contains mostly irregular or fusiform small neurons differing from those in adjacent structures by their size, and by their intensity of staining with in situ hybridization and immunohistochemistry. The connections of this nucleus (which is called here the bed nucleus of the stria medullaris, BSM) are unclear, but evidence in the literature suggests that it may receive inputs from the fornix and project through the stria medullaris to the medial habenula. PMID- 8719274 TI - Light-microscopic distribution and parasagittal organisation of muscarinic receptors in rabbit cerebellar cortex. AB - Recent studies on the effects of intrafloccular injections of muscarinic agonists and antagonists on compensatory eye movements in rabbit, indicate that muscarinic receptors may play a modulatory role in the rabbit cerebellar circuitry. It was previously demonstrated by Neustadt et al. (1988), that muscarinic receptors in rabbit cerebellar cortex are distributed into alternating longitudinal zones of very high and very low receptor density. In the present study, the zonal and cellular distribution of muscarinic receptors in the rabbit cerebellar cortex is investigated in detail using in vitro ligand autoradiography with the non selective high-affinity antagonist [3H]quinuclidinyl benzilate (QNB), and the M2 specific antagonist [3H]AF-DX384, and immunocytochemistry with a monoclonal antibody specific for the cloned m2 muscarinic receptor protein. [3H]QNB and [3H]AF-DX384 binding sites and m2-immunoreactivity had similar overall distributions: dense labeling occurred in the dendritic arbors of a subset of Purkinje cells that are organized into parasagittal bands. A high level of muscarinic receptor labeling was also observed in a thin substratum of the molecular layer immediately above the Purkinje cell layer of the vestibulo cerebellar lobules, i.e. the nodulus, the ventral uvula and the flocculus. Labeling in this stratum was associated with densely packed fibres, which were putatively identified as parallel fibres. Also Golgi cells, which were localized in part in the molecular layer, and a subset of mossy fibre rosettes, primarily concentrated in lobule VI, were immunoreactive for the m2 receptor. The parasagittal band of labeled Purkinje cell dendrites were most prominent in the anterior lobe (lobules I-V), in crus 1 and 2, in the flocculus, the ventral paraflocculus and the rostral folium of the nodulus. In other lobules, only infrequent Purkinje cells contained muscarinic receptors. The parasagittal organisation of muscarinic receptors differed from that of zebrin I, a Purkinje cell-specific protein which is often used as a marker of parasagittal parcelation of the cerebellar cortex. In the anterior lobe, however, there was a partial correspondence between muscarinic receptor and zebrin I bands. In the flocculus the distribution of muscarinic-receptor-positive Purkinje cells was related to the distinct white matter compartments as revealed with acetylcholinesterase (AChE) histochemistry. Muscarinic receptor-containing Purkinje cells were located primarily in the floccular zone 1, which is implicated in the control of eye movements about a horizontal axis. In order to relate the distribution of muscarinic receptor labeling to that of cholinergic nerve terminals, [3H]QNB binding sites and sodium-dependent [3H]hemicholinium-3 binding were compared. Sodium-dependent [3H]hemicholinium-3 binding sites mainly occurred in the granule cell layer of the vestibulo-cerebellum, which corresponds well with the distribution of the acetylcholine synthesizing enzyme, choline acetyltransferase (ChAT). However, sodium-dependent [3H]hemicholinium binding complemented, rather than co-localized with, muscarinic receptors which were primarily distributed in the molecular layer of the lobules of the vestibulo-cerebellar lobules. Their functional significance is puzzling, since their distribution does not correspond to that of markers of cholinergic innervation. PMID- 8719275 TI - Glutamate-like immunoreactivity in synaptic terminals of the posterior cingulopontine pathway: a light and electron microscopic study in the rabbit. AB - A postembedding immunoperoxidase method for light microscopy was used to localize glutamate-like immunoreactivity in the rabbit basilar pontine nuclei. Labelled fibre bundles, neuronal cell bodies and numerous puncta of diverse size were heavily glutamate immunoreactive throughout all subdivisions of the pontine nuclei. To determine whether some of the glutamate-immunoreactive puncta were synaptic terminals of posterior cingulate cortical neurons, a double-labelling technique involving an anterograde tract-tracing method and a postembedding immunogold procedure for electron microscopy was used. A quantitative evaluation of gold particle densities revealed that anterogradely labelled cingulopontine synaptic terminals were about twice as immunoreactive as their postsynaptic dendrites, perikaryal and glial profiles and about three times more than symmetric synaptic terminals. The present results indicate that the posterior cingulopontine projection contains high levels of glutamate at its synaptic terminals. This observation provides further support to the role for glutamate as a neurotransmitter in the corticopontine pathway. PMID- 8719276 TI - Ontogeny of GABA-immunoreactive neurons in the central nervous system in a teleost, gasterosteus aculeatus L. AB - The inhibitory neurotransmitter gamma-aminobutyric acid (GABA) is known to exert various neurotrophic actions in the developing nervous system, but little is known about its distribution in the central nervous system during early development. We have studied the development of GABA-immunoreactive (GABAir) neurons during embryogenesis of a teleost fish, the three-spined stickleback. As early as 51 h postfertilization (PF; hatching occurs 144-168 h PF, and the first monoaminergic neurons appear around 72 h PF) GABAir neurons appear in the ventral prosencephalon caudal to the optic recess, in the ventral mesencephalon, and in the spinal cord. Then, there is a gradual addition of GABAir cell groups in the rostral prosencephalon and ventral rhombencephalon (66 h PF), dorsal and caudal hypothalamus and pretectum (72 h PF), ventral hypothalamus (78 h PF), preoptic region, thalamus, and in the mesencephalon and rhombencephalon (96 h PF). GABAir axons appear in the spinal cord already at 51 h PF, and then gradually appear in the various tracts of the early axonal scaffold of pathfinding fibers, so that by 96 h PF the entire axonal scaffold contains GABAir fibers. It appears likely that GABAergic axons contribute a major population to the formation of the axonal scaffold. Moreover, in the prosencephalon GABAir neurons are arranged in clusters that may reflect a neuromeric organization with six prosencephalic neuromeres. PMID- 8719277 TI - Neurotransmitter and neuromodulator systems of the rat inferior colliculus and auditory brainstem studied by in situ hybridization. AB - This study was concerned with the distribution of a variety of putative neuromodulator and neurotransmitter systems in auditory regions of the rat brainstem using in situ hybridization histochemistry. Serial brain sections were screened for the presence of mRNAs for (i) precursors of the neuroactive substances cholecystokinin, somatostatin, proenkephalin and substance P (preprotachykinin), (ii) glutamic acid decarboxylase, the key synthesizing enzyme for GABA, or (iii) subunits alpha 1, alpha 2 and alpha 3 of the GABAA receptor. Detectable message for all of these probes was found in at least one auditory brainstem area. There were clear differences in the distribution of the various mRNAs in subregions of the inferior colliculus, superior olivary complex, lateral lemniscus and cochlear nucleus. Cells expressing mRNA for glutamic acid decarboxylase were most prominent in the inferior colliculus, but were also present in all lower auditory brainstem nuclei, except the medial superior olivary nucleus and medial nucleus of trapezoid body. The mRNA for GABAA alpha 1 receptor subunits was detectable in all auditory regions investigated, although at different levels of expression. GABAA alpha 2 and alpha 3 mRNA signals were seen in inferior colliculus, lateral lemniscus and in almost all superior olivary complex regions, but in fewer cells and at lower levels than the GABAA alpha 1 subtype. Moderate to high levels of preprocholecystokinin mRNA expression were seen in all subregions of the inferior colliculus. In other auditory brainstem areas, preprocholecystokinin mRNA levels were either low or absent. With regard to mRNAs for the neuroactive peptides somatostatin, preprotachykinin and preproenkephalin, all were expressed in the inferior colliculus but there were differences in their cellular distribution. For example, there were almost no preprotachykinin mRNA expressing cells in the central nucleus of inferior colliculus and levels of somatostatin mRNA were especially high in the dorsal cortex and in layer 3 of the external cortex of inferior colliculus. There were also differences in the pattern of expression of these mRNAs in the various brainstem auditory nuclei; there was no preprotachykinin mRNA in any part of the superior olivary complex, only somatostatin mRNA was found in the ventral cochlear nucleus, and expression of preproenkephalin mRNA was pronounced in the ventral nucleus of the trapezoid body and the rostral periolivary zone. The data are considered in light of the connectivity and functional organization of the auditory brainstem. PMID- 8719278 TI - Albinism and the associated ocular defects. AB - Several types of hypopigmentation in humans are called albinism. The phenotype for different types of albinism varies according to the amount of pigment in the hale, skin and iris, the reduction in visual acuity and the degree of nystagmus and strabismus. Cutaneous and ocular melanin pigment can range from complete absence throughout the lifetime of the individual to the development of nearly normal levels, including the ability to tan. Visual acuity ranges from 20/40 to 20/400, and visual development in an affected infant is slower than normal. Foveal hypoplasia and altered routing of the optic nerves are found in all types of albinism and are the most constant feature of this condition. The demonstration of optic track misrouting by visual evoked potential studies provides the critical diagnostic procedure for questionable cases of albinism, and this is the single definitive diagnostic test to confirm a diagnosis of albinism. PMID- 8719279 TI - The course of nonproliferative diabetic retinopathy following ECCE with posterior chamber IOL implantation. AB - A prospective study on the effect of ECCE + IOL implantation on the course of diabetic retinopathy (DR) in 59 eyes showed that in one to three years following surgery, progression of DR, including the development of newly formed one, occurred in 28.8% of the eyes. This was more marked in patient with IDDM (IDDM 46%, NIDDM-24%) as well as in eyes with pre-operative BDR (pre-operative BDR 62.5%, no pre-operative BDR-16.3%). Final visual acuity was better in eyes without pre-operative DR as well as in eyes that did not show progression of the retinopathy. PMID- 8719280 TI - Evaluation of the results of intraocular lens implantation in Egyptian children. AB - A retrospective study was done for 95 eyes of Egyptian children in whom I.O.L. implantation was performed between 1985 and 1986 to treat unilateral cataract due to trauma or congenital or aphakia. The average range of follow up period was three to four years old. It was revealed that I.O.L. In traumatic cataract gave the best results (as regards the visual acuity with minimal complications) especially with anterior chamber lenses and still the secondary implantation of A.C. lenses had to some extent better results than that of primary A.C. lenses. In congenital cataract, P.C. lenses gave the worst results. Again, the older the age of the children, the better the results of I.O.L. implantation. PMID- 8719281 TI - Evolution and natural history of diabetic retinopathy during pregnancy. AB - The metabolic stress in both pregnancy and diabetic mellitus being endocrine mediated, it has always been feared whether a patient with an underlying retinopathy will progress into the more difficult proliferative stage with the coming of pregnancy. Admittedly, this important question has been answered differently by different investigators. However, much of these studies has been only impressive statistical analysis of large number of cases without any laboratory or clinical investigations being undertaken alongside to ascertain into the role of all endocrine hormones. We have tried to fill this omission here in this study. From this study we have come to believe that there is no factual evidence of deleterious relationship between the onset of pregnancy and the underlying diabetic retinopathy. Matter of factly, our study show that pregnancy may actually provide respite from developing further complications. Here, the hormone prolactin may play an important protective role. PMID- 8719282 TI - Genotypic evaluation of ocular pathologies in sickle cell diseases. AB - Sickle cell disease patients are known to manifest different types of ocular problems. These problems include proliferative and non-proliferative retinopathies, and refractive errors. The distribution of these pathologic and refractive problems among the sickle cell genotypes is analyzed according to the individual genotypes. Data collected from a total of 63 sickle cell disease patients who responded to the questionnaires shows that fifty-six percent (35) are males and forty-four percent (28) are females. The genotype distributions are: SS, 55% (34); SC, 33% (21); S-Thal, 11% (7); while 2% (1), is AS-genotype. Overall findings show that severe ophthalmic abnormalities were reported by approximately 90% of the respondents with sickle cell disease. The majority of sickle cell disease patients in this research show consistent susceptibility to both fungal and bacterial infections with varying degree of refractive errors. It is observed that refractive errors are prevalent in SC and S-Thal sickle cell genotypes, while the severity of anemia and painful crises are more prevalent in SS and S-Thal genotypes. Systemic ocular problems are also prevalent among the SC and S-Thal disease genotypes. PMID- 8719283 TI - Influence of systemic diseases and environmental factors on age at appearance, location and type of acquired cataract. AB - We studied 257 patients scheduled for cataract surgery for possible correlation with systemic diseases, blood chemistry, and environmental exposure. We found that posterior subcapsular cataract appeared in patients 10 years younger on average than those with other types of cataract. Nuclear cataract patients had higher blood levels of uric acid and creatinine and lower blood levels of calcium. We suggest capsular insult as a possible pathophysiological explanation for cataract formation in young age groups (as well as diabetes) and a toxic effect of blood metabolites in patients with renal failure, resulting in earlier and more prevalent sclerosis of lens fibers. No effect of smoking or of exposure to sunlight on the distribution of various types of cataracts was noted. PMID- 8719284 TI - Metabolic disturbance in age-related macular degeneration. AB - The photodynamic effect may damage the choroid-retina in AMD, which involve to oxygen producing the superoxide free radical. The enzymatic and non-enzymatic defence systems would appear the capacity to scavenge the toxication in choroid and retina. We observed the superoxide dismutase and catalase decreased as well as the decrease of serum zinc in AMD patients than in the controls (p > 0.01). The results showed the protective capacity decreased by the enzymatic and non enzymic defense systems in some AMD patients. It may be one of the important roles in the pathogenesis of AMD. PMID- 8719285 TI - Nutrition and metabolic eye disease. AB - Certain metabolic eye disorders may be caused by nutritional factors, and others may be treated by nutritional adjustments or withdrawal of certain elements in the diet, and sometimes the inclusion of certain drugs. Occasionally a metabolic eye disease is drug induced. Quite often, certain addition or subtraction to or from the patient's regimen may influence the progress of the metabolic eye disease. PMID- 8719286 TI - Whether to measure prothrombin time at the ambulatory care pharmacy. PMID- 8719287 TI - HMO pharmacist in a contract hospital. PMID- 8719288 TI - Smoking's challenge to pharmacy. PMID- 8719289 TI - Pharmacoeconomic analysis of smoking-cessation interventions. AB - A model for evaluating the costs and benefits of smoking-cessation interventions is proposed and published data on the subject are analyzed. Decision-tree analysis was used to determine the costs and benefits, from an employer's perspective, of several smoking-cessation options. The probability that a smoker would stop smoking was based on two published meta-analyses of smoking-cessation interventions and a national study of nicotine patch users who received a smoking cessation consultation from a pharmacist. The costs were determined by telephone interviews and market research information. A sensitivity analysis of treatment costs was conducted to determine the optimal treatment from the standpoint of costs versus benefits. The smoking-cessation intervention from which an employer would receive the greatest net benefit is the nicotine patch with smoking cessation consultation from a pharmacist and patient participation in a formal smoking-cessation program. Pharmacists included in this analysis could charge a smoking-cessation consultation fee of up to $109 per patient. Pharmacoeconomic analysis suggested that smoking-cessation therapy consisting of treatment with nicotine patches, consultations with pharmacists, and participation in a comprehensive behavioral program would be the most cost-beneficial to employers. PMID- 8719290 TI - Disease management programs. AB - Disease management (DM) activities are described, and their implementation and monitoring in managed care organizations are discussed. DM programs involve systematic evaluation of the relationships between treatment options and the associated resource use and patient outcomes for the purpose of providing a given standard of health care at the lowest possible resource cost. A DM arrangement covers a specified disease or therapy intervention for a patient group that may be defined by diagnosis, drug use, prior resource use, or patient characteristics. Often, the partners in a DM arrangement are a managed care organization and a pharmaceutical industry representative or division. The development and monitoring of disease management arrangements are dependent on access to several types of data, and these data are available in managed care plans. A DM arrangement includes interventions to change prescribing patterns or patient compliance and assessment of the effects of these interventions against target outcomes specified in the contract. The agreement that is developed specifies guidelines for treatment and requirements for data collection, monitoring, and reporting that are consistent with the target outcomes. In many DM arrangements, the partners share cost savings and risk; other arrangements involve case management on a capitated basis. A pharmaceutical company involved in risk sharing must change its focus from market share to optimal use of drugs within the total cost of treatment. If a risk-sharing contract covers an entire therapeutic class of drugs, a pharmaceutical company may share risk for the use of other manufacturers' products as well as its own. Disease management contracts must consider the full impact of each treatment option on the health system; the goal should be not simply to decrease the drug budget, but to decrease overall costs for treatment that achieves desired outcomes for specific diseases. PMID- 8719291 TI - ASHP supplemental standard and learning objectives for residency training in pharmacotherapy practice. The American College of Clinical Pharmacy. American Society of Health-System Pharmacists. PMID- 8719292 TI - The economics of smoking cessation. PMID- 8719293 TI - The keyboard and the Camel. PMID- 8719294 TI - Concentration-dependent precipitation of sodium bicarbonate with ciprofloxacin lactate. PMID- 8719295 TI - Preparing a hospital-based home care service for Joint Commission evaluation. PMID- 8719296 TI - Mislabeled cherry syrup. PMID- 8719297 TI - Regulatory effect of 1,25-dihydroxyvitamin D3 on insulin release and calcium handling via the phospholipid pathway in islets from vitamin D-deficient rats. AB - The effect of 10(-8) M 1,25-dihydroxyvitamin D3 [1,25 (OH)2D3] on the phosphoinositide pathway, was studied on [3H] inositol and 45Ca2+ efflux and on insulin release of islets from vitamin D-deficient rats, during an acetylcholine (Ach) stimulus in perifusion. The insulin release, which was low in vitamin D deficient rats, was enhanced by this treatment. The 3H flux, reflecting phosphoinositide breakdown, was also increased. The 45Ca2+ flux was stimulated both during the first 14 min peak (mobilization of IP3-sensitive reticular Ca2+ stores) and during the following sustained small elevation of 45Ca2+ flux, reflecting protein kinase C (PKC) activation and consequently increased phosphorylation of Ca2+ channel proteins. These effects were larger during perifusions performed in the presence of glucose which is known to open Ca2+ channels, suggesting a synergistic influence of glucose and 1,25(OH)2D3. This positive influence of 1,25(OH)2D3 in Ca2+ entry by Ca2+ channels was confirmed by the use of nifedipine-a Ca2+ channel blocker-which suppressed the 45Ca2+ flux and lowered insulin secretion. Moreover, the sustained 45Ca2+ flux also disappeared in islets from vitamin D-deficient rats supplemented by 1,25(OH)2D3 but perifused without extracellular Ca2+ supporting the hypothesis of 1,25(OH)2D3-induced activation of PKC. Thus, 1,25(OH)2D3 may provide supplementary calcium to the B cell by regulating the intracellular signalling processes involving phospholipid metabolism, PKC induction, Ca2+ mobilization and Ca2+ entry by Ca2+ channels. PMID- 8719298 TI - Na+, K+ ATPase activity in red cells predicts the recurrence of hyperthyroidism in patients with Graves' disease. AB - Graves' disease is an autoimmune disorder characterized by a course of remission and relapse. Several parameters have been evaluated for their abilities to predict the clinical course of Graves' disease in patients treated with antithyroid drugs. We recently demonstrated in patients with hyperthyroidism dependent by Graves' disease, an impaired Na+, K+ ATPase activity in red cells and a correlation between ATPase and free T3. With the aim to clarify the relationship between the course of hyperthyroidism and the Na+, K+ ATPase activity during and after discontinuing the antithyroid therapy, we followed up 24 patients for two years. In our previous work by restoring a normal level of free T3, we obtained a normalization of Na+, K+ ATPase activity in the red cells of all the patients. However, in eight subjects after a period of 150 days following the suspension of therapy, we observed a new reduction of ATPase activity in a clinical condition of euthyroidism. The same subjects, newly evaluated after 150 days, developed a clinical and biochemical relapse of hyperthyroidism. We believe that the determination of Na+, K+ ATPase activity is able to predict the recurrence of hyperthyroidism in patients with Graves' disease. PMID- 8719299 TI - Effect of testosterone replacement therapy on the somatotrope responsiveness to GHRH alone or combined with pyridostigmine and on sympathoadrenal activity in patients with hypogonadism. AB - There is evidence suggesting that androgens influence GH secretion in man. Our aim was to verify whether the GH releasable pool is preserved and influenced by testosterone replacement in male hypogonadism. To this goal, in eight male hypogonadal patients (HP, age 32.2 +/- 5.0 yr; Body Mass Index 23.9 +/- 1.1 kg/m2) before and after 3 months testosterone therapy, we studied the GH response to GHRH (1 microgram/kg iv) alone and combined with pyridostigmine (PD, 120 mg po), a cholinesterase inhibitor which likely inhibits hypothalamic somatostatin release allowing exploration of the maximal somatotrope secretory pool. Sixteen normal subjects (NS, age 30.1 +/- 3.5 yr; Body Mass Index 22.5 +/- 1.8 kg/m2) were studied as controls. The GH response to GHRH in HP was similar to that in NS (AUC, mean +/- SE: 1238 +/- 362 vs 1018 +/- 182 micrograms/L/h). PD potentiated to the same extent the GH response to GHRH in both groups (2092 +/- 807 and 2840 +/- 356 micrograms/L/h). After three month testosterone therapy, in HP the GH responses to GHRH alone (1352 +/- 612 micrograms/L/h) and combined with PD (1948 +/- 616 microgram/L/h) were unchanged. Also IGF-I levels in HP were similar to those in NS (222 +/- 42 vs 210.6 +/- 55.8 micrograms/L) and were unchanged during testosterone replacement (280 +/- 31 micrograms/L). As androgens have been reported to modulate sympathoadrenal activity in the rat, both before and during testosterone replacement, we also measured plasma catecholamine levels. Basal NE (p < 0.05) but not E levels were lower in HP than in NS; testosterone restored basal NE levels to normal without affecting basal E. delta absolute increase of NE and E (p < 0.05 and 0.01 vs baseline, respectively) after PD in HP were similar to those in NS and were unchanged during testosterone replacement. In conclusion, these results demonstrate that the GH releasable pool is preserved in male hypogonadism. As in this condition a reduction of spontaneous GH secretion has been reported, it could be due to neurosecretory dysfunction but not to pituitary impairment. Subtle alterations of sympathoadrenal activity seem to be present in male hypogonadism and reversed by testosterone replacement. PMID- 8719300 TI - Morning plasma free cortisol: inability to distinguish patients with mild Cushing syndrome from patients with pseudo-Cushing states. AB - Patients with Cushing syndrome have elevated 24 h urinary free cortisol excretion and loss of diurnal rhythm of plasma cortisol, however, morning total plasma cortisol concentrations are often similar to those of normal individuals. To test the hypothesis that an elevated morning plasma free cortisol could distinguish patients with Cushing syndrome from those without the disorder, we measured plasma free cortisol levels at 08:00 in 20 patients with surgically-confirmed mild Cushing syndrome, 19 patients with pseudo-Cushing states, and 9 normal volunteers. We then determined the sensitivity, specificity and diagnostic accuracy of plasma free cortisol for the diagnosis of hypercortisolism and for the diagnosis of Cushing syndrome. Plasma free cortisol was compared to 08:00 total plasma cortisol and to basal 24 hour urinary free cortisol. Morning plasma free cortisol was significantly elevated in patients with mild Cushing syndrome (58 +/- 6 nmol/L; mean +/- SE) compared to plasma free cortisol in patients with pseudo-Cushing states (36 +/- 6 nmol/L) (p < 0.005) and normal volunteers (22 +/- 4 nmol/L) (p < 0.0005). Total morning cortisol was significantly greater in patients with mild Cushing syndrome (470 +/- 28 nmol/L) than in normal volunteers (310 +/- 33 nmol/L, p < 0.05) but was not different from patients with pseudo Cushing states (410 +/- 47 nmol/L, p = NS). Urinary free cortisol was also significantly greater in mild Cushing syndrome (540 +/- 50 nmol/d) than in patients with pseudo-Cushing states (430 +/- 41 nmol/d, p < 0.05) or normal volunteers (160 +/- 28 nmol/d, p < 0.0001). However, there was considerable overlap of plasma free cortisol, plasma total cortisol and urinary free cortisol between the three groups precluding these tests alone from being useful to distinguish patients with mild Cushing syndrome from patients with pseudo-Cushing states. To achieve 100% sensitivity for the diagnosis of Cushing syndrome, the specificity for plasma free cortisol, plasma total cortisol and urinary free cortisol was 0.11, 0.32 and 0.32, respectively. We conclude that plasma free cortisol is of limited value in the differential diagnosis of hypercortisolism. PMID- 8719301 TI - Effect of triiodothyronine administration in experimental myocardial injury. AB - Twelve healthy pigs were subjected to a 20-min, period of regional myocardial ischemia by snaring the left anterior descending coronary artery (LAD) between its first and second diagonal branches. The resulting myocardial injury caused significant acute hemodynamic impairments. Cardiac index declined significantly during reperfusion interval and returned to preischemic level by postoperative day 7. Plasma total triiodothyronine (TT3), free triiodothyronine (FT3) and free fatty acid (FFA) decreased gradually and reached the nadir at 6 h after LAD occlusion. In contrast, plasma reverse triiodothyronine (rT3) increased progressively after LAD occlusion and reperfusion. To investigate the effect of T3 on ischemic myocardium, T3 (0.2 microgram/kg/dose; n = 5) or saline (placebo; n = 6) was administered immediately, 30 min, 60 min, 90 min, and 120 min after reperfusion. Plasma TT3 and FT3 increased dramatically after triiodothyronine supplement but declined to presichemic level at six h after LAD occlusion. The pigs treated with T3 demonstrated a rapid improvement in cardiac index over the reperfusion interval, whereas cardiac index in the placebo group remained depressed. Myocardial oxygen consumption estimated by rate pressure product showed no difference between placebo and T3-treated groups. Oxygen extraction as O2 saturation difference between aorta and coronary sinus was less in T3-treated group. Nine pigs (four in the T3-treated group and five in the placebo group) were subjected to euthanasia with hypertonic KCl solution on postoperative day 7. Myocardial infarct size determined by triphenyltetrazolium chloride (TTC) tissue enzyme staining technique was not significantly different between T3-treated and placebo groups. We concluded that this animal model is a useful model of myocardial injury simulating "euthyroid sick syndrome" as seen in patients with cardiopulmonary bypass, and T3 supplementation after reperfusion significantly enhanced postischemic left ventricular functional recovery but did not affect myocardial oxygen consumption and myocardial infarct size. PMID- 8719302 TI - Statistical analysis of functional region(s) of the TSH receptor. AB - In this study, we apply recently developed methods to evaluate the thyrotropin receptor (TSH-R). These methods are called deviation (DEV) model, deviation decrease (DD) and DEV/DD analyses, and are based on deviation of amino acid sequences. A 3-dimensional structure model of TSH-R was graphically constructed, and found to possess a large central cavity (donut-like structure). The N terminus was found to be in the center of the whole extracellular structure and to form a part of the bottom of the cavity. High DEV values indicate deviated amino acid compositions in the protein and were seen in 7 regions, 6 of which were found to be in regions with hydrophilic and acrophilic character. On the basis of the analysis of intra-molecular cis-acting relationships, 7 pairs of regions were presumed to be closely related. Further, when 3 exoplasmic loop lesions were analyzed similarly, 3 other regions were shown to have a close relationship with the cell surface. DEV/DD values were applied to predict the interface of TSH-R with trans-acting molecules such as TSH-R antibody or TSH. The regions in association with trans-acting molecules were seen in 14 regions, 11 of which included the high DEV regions. Both of the TSH-R specific regions in the N- and C-terminal side, especially the latter, were found to be the major components. PMID- 8719303 TI - GH responsiveness to repeated GHRH or hexarelin administration in normal adults. AB - GH responses, calculated as the net incremental area under the curve (GH nAUC/h), to two consecutive 1 microgram/kg/bw iv GHRH boluses (administered at 0 and 120 min, test a), to one 1 microgram/kg/bw iv GHRH bolus followed by a 1 microgram/kg/bw iv hexarelin bolus (administered at 0 and 120 min respectively, test b) and to two consecutive 1 microgram/kg/bw iv hexarelin boluses (administered at 0 and 120 min, test c) were evaluated in 6 normal adults. The first GHRH injection caused a clear rise in serum GH levels in all subjects (mean GH nAUC/h, test a: 832.1 +/- 59.4 ng/ml/h, range: 723.7-1074.0 ng/ml/h; test b: 859.2 +/- 122.9 ng/ml/h, range 618.0-1422.7 ng/ml/h). Hexarelin administration elicited a marked GH release (test c: 1424 +/- 208.2 ng/ml/h, range: 810.0-2154.0 ng/ml/h), which was significantly higher than those observed after GHRH (vs test a: p < 0.02, vs test b: p < 0.05). After the first GHRH bolus, the second GHRH injection (test a) was unable to sustain GH elevated levels (mean GH nAUC: 74.5 +/- 26.5 ng/ml/h, range: 9.7-182.2 ng/ml/h), while hexarelin administration (test b) caused a clear GH rise in all subjects (GH nAUC: 1049.7 +/- 105.2 ng/ml/h, range: 786.0-1356.0 ng/ml/h). Repeated hexarelin administration (test c) was associated with a significant (p < 0.02) reduction of GH responses to the second bolus (286.6 +/- 43.1 ng/ml/h), which were however significantly higher than those observed after the second GHRH bolus (p < 0.05). In conclusion, these results demonstrate that repeated hexarelin administration causes a reduction of GH responsiveness, which is less marked than that induced by repeated GHRH administration, probably due to the more potent GH-releasing effect of hexarelin. Moreover, when administered after GHRH, hexarelin has a sustained GH-releasing effect in contrast to the poor efficacy of GHRH, thus suggesting that the acute effect of hexarelin is probably not mediated through hypothalamic GHRH pathways. PMID- 8719304 TI - Temporal relationship between bone loss and increased bone turnover: a longitudinal study following natural menopause. AB - We report the results of a longitudinal study aimed at better defining concomitant changes of both bone mineral density (BMD) and of four independent markers of bone turnover (serum osteocalcin, serum alkaline phosphatase activity, fasting urinary hydroxyproline/creatinine and calcium/creatinine ratio) following natural menopause. The results obtained indicate that, within a relatively short period of time since cessation of gonadal function, conventional markers of bone turnover behave differently. In fact, while the mean values of hydroxyproline/creatinine ratio (felt to be a marker of bone resorption) rise immediately at the first control (19.7 +/- 11.7 months), the bone formation markers gradually increase and, as far as serum osteocalcin levels are concerned, this increment appears to be long-lasting. As a result of these changes, a negative skeletal balance follows, which is documented by the prolonged reduction of bone mineral density during the entire observation period. Mean +/- SD % measured yearly bone loss was -2.83 +/- 2.6. There was a highly significant correlation between initial and final BMD values (r = 0.908, p < 0.001; r2 = 82.5) and a weak inverse correlation (r = -0.298, p < 0.046) between initial serum alkaline phosphatase values and % yearly bone loss. In conclusion, measurement of the biological indices of bone remodelling following natural menopause indicate that the increase in osteogenesis is delayed compared to that of bone resorption; furthermore, in the immediate postmenopausal period, the actual bone mass should be considered the best predictor of future bone mass. The inverse correlation found between % yearly bone loss and serum alkaline phosphatase values seems to emphasize the importance of increased bone turnover as an independent predictor of bone loss. PMID- 8719305 TI - Isolated TSH deficiency with a partially empty sella. AB - Isolated TSH deficiency is rare. The diagnosis is based on (i) symptoms and signs of thyroid hormone deficiency, (ii) demonstration of an absent or impaired TSH response to TRH and (iii) other pituitary hormones remaining intact. We report a 60-year-old female in whom isolated TSH deficiency was found, with a free thyroxine - 7.0 pmol/L (11-20), total triiodothyronine level - 1.5 nmol/L (1.1 2.6) and thyroid stimulating hormone - 0.87 mU/L (0.38-4.3). A TRH test on two separate occasions demonstrated an inappropriately low TSH response. Computed assisted tomography revealed a partially empty sella and other pituitary hormones were demonstrated to be intact. We were unable to detect pituitary antibodies using indirect immunofluorescence on sections of monkey pituitary. Treatment with thyroxine improved this patient's symptoms and suppressed the TSH further. PMID- 8719306 TI - IGFs in the feedback control of GH secretion: Hypothalamic and/or pituitary action? PMID- 8719308 TI - The kerosene tragedy of 1994, an unusual epidemic of burns: epidemiological aspects and management of patients. AB - An unusual and perhaps the first epidemic of burns occurred between 15 February 1994 and mid April 1994 in four districts of the State of Rajasthan in India. The cause of this epidemic was the accidental mixing of petrol in kerosene oil which was inadvertently overlooked. This mixture of kerosene and petrol was used mainly by people of low-income groups for lighting lamps. Most of the accidents occurred while pouring this highly inflammable petrol-kerosene mixture into ignited lamps. A total of 303 cases were reported: 118 of these patients sustained severe burns of whom 37 died. Small numbers of fresh cases kept occurring over a period of 2 months in spite of all efforts by the administration, because poor people kept using the fatal mixture due to ignorance and illiteracy. Most of the patients were managed at district hospitals with the help of plastic surgeons called for the purpose from Jaipur, the capital city of the affected State. A total of 40 out of 303 patients were transferred to SMS Hospital where a medical ward was vacated to manage these patients, as the 10-bed burn unit already had 300 per cent best occupancy. Most of these patients were not willing to be sent to a burn unit situated far away from their homes, but they had to be transferred because the general surgeons working at district hospitals were hesitant to manage them, not so much due to lack of training in the management of burns, but more due to lack of willingness to manage burns. This indicates the need for renewed emphasis not only of the necessity of training general surgeons, nursing and paramedical staff at district level in the management of burns, but also of the need to manage these cases at district level. This idea needs serious consideration and sincere efforts to implement it at the national level. The paper has been split into two parts: epidemiological aspects and management of patients. PMID- 8719309 TI - Acceleration of skin graft healing by growth factors. AB - A special local burn wound treatment, using a healing promotion factor, attempted to achieve earlier wound closure. Clinically it is not difficult to harvest autologous human platelet-derived wound healing factor (HPDWHF) from burn patients. Another study has proved that porcine pituitary extract (PPE) could be one of the supplements for growing human keratinocytes in vitro (Wang HJ, Chen TM, Tung YM et al. Burns 1995). Initially, using the ELISA immunoassay, we detected titres of platelet-derived growth factor heterodimer AB (PDGF-AB) and epidermal growth factor (EGF) in HPDWHF and PPE. After elevating a 6 X 9 cm2 flap on the backs of Sprague-Dawley (SD) rats 1 ml of heterologous HPDWHF and PPE were sprayed topically on the wound and followed by grafting six pieces of 1 cm2 autologous full-thickness skin. The flap wound was then closed with a piece of large-pore Biobrane interposed between the skin graft wound and the flap. On postgraft day 7, the wound was reopened to measure the area of the full-thickness skin graft (FTSG) using micrographic paper; the results are reported as the percentage of graft expansion beyond the original size. In the cytokine study, we found that the concentrations of PDGF-AB dimer in HPDWHF and PPE were 5222 +/- 102 and 375 +/- 12 pg/ml (both values are reported after a 1:10 dilution) (n = 3), whereas the EGF concentration was 245 +/- 9 pg/ml, and undetectable in undiluted PPE (n = 3), thereby proving that the PDGF-AB and EGF fibres in the HPDWHF were significantly higher than in the PPE Both the HPDWHF (n = 6, with each animal grafted with six skin graft samples) and PPE (n = 7) demonstrated significant accelerations of FTSG growth, with 14.41 +/- 1.08 per cent and 13.16 +/- 3.25 per cent increases in the FTSG size when compared to the sham control group 1.39 +/- 3.26 per cent (n = 5) (P < 0.05). Comparisons between the treatment groups showed no evidence to indicate that the HPDWHF is superior to the PPE in accelerating FTSG growth (P > 0.05). This supports the hypothesis that PDGF appears to transduce its signal through wound macrophages and may trigger the induction of positive autocrine feedback loops and synthesis of endogenous wound PDGF and other growth factors, thereby enhancing the cascade of tissue repair (Pierce GF, Mustoe TA, Altrock B et al. 1991; J Cell Biochem 1991; 45: 319 326), and the effect that PDGF-AB on wound healing is not dose dependent (Mustoe TA, Cutler NR, Allman RM et al. Arch Surg 1994; 129: 213-219). In future studies we expect to show that autologous HPDWHF and heterologous PPE will promote skin graft wound healing in burn patients since they contain a high concentration of PDGF-AB. PMID- 8719310 TI - Bacterial translocation in D-galactosamine-treated rats in a burn model. AB - Bacteria and endotoxins can pass through the gut barrier under certain conditions. This process of bacterial translocation (BT) may occur after thermal injury in animals and is thought to play a role in the pathogenesis of septic complications in severely burned patients. The current study was performed to determine the role of endotoxin-related cytokines in the pathogenesis of burn induced BT. Wistar rats were used in which enhanced sensitivity to TNF/LPS reactions was achieved by treatment with galactosamine (GalN). The GI tracts of these rats were antibiotic decontaminated with oral bacitracin and neomycin and were colonized with a neomycin resistant (NR)-Escherichia coli strain. The rats were divided into four groups, 30 per cent TBSA scald with GalN (Burn+GalN) pretreatment; 30 per cent TBSA scald without GalN (Burn); or sham injury with (GalN) and without GalN (Sham) pretreatment. On day 2, the animals were killed and liver, spleen, lung, heart and the peritoneal cavity were cultured. Blood samples were taken and the concentrations of LPS, TNF, IL-6 and ALAT were determined. Mortality was significantly increased in the Burn+GalN group compared to the other groups. In all groups, the incidences of BT were increased compared to the sham-treated group, although BT was most pronounced in the Burn+GalN group. In the latter group it was accompanied by highly elevated IL-6 and ALAT levels. The results of this study suggest that endotoxin mediators like TNF and IL-6 could play a role in the phenomenon of BT and that the function of the liver is an important clearing mechanism. PMID- 8719311 TI - Successful 14-day preservation of rat skin using a two-layer cold-storage method. AB - Skin-graft preservation for the purpose of delayed autografting to burn patients is a basic tool of plastic and burn surgery. This study was undertaken to evaluate the efficacy of the two-layer (physiological saline/perfluorochemical (PFC)) cold-storage method for skin preservation. In Wistar rats, a full thickness skin graft, 3 cm in diameter, was harvested from the back, and separated from the dermis to obtain the epidermal sheet. This epidermal sheet was preserved at 4 degrees C using physiological saline (group 1, n = 15), physiological saline bubbled with a 95 per cent oxygen, 5 per cent carbon dioxide mixture (group 2, n = 15) or a two-layer method with bubbling with 95 per cent oxygen and 5 per cent carbon dioxide mixture (group 3, n = 15). The epidermal skin sheets were then autotransplanted to the backs of the original donor rats 7, 10 and 14 days after the start of preservation. The success rate of the skin autotransplantation was determined by measuring the ratio of viable area to total graft area 7 days after grafting. In group I, the success rates after 7, 10 and 14 days' perservation were 56.4, 47.4 and 0.09 per cent, respectively. In group 2, the corresponding success rates were 62.0, 48.4 and 0.8 per cent respectively, showing no improvement with oxygenated saline. In clear contrast, the success rates were significantly improved by the two-layer method, the values being 92.1, 87.9 and 77.6 per cent after 7, 10 and 14 days of perservation, respectively (P < 0.01 vs. groups 1 and 2 for all preservation durations). Based on these results, we concluded that the two-layer method is useful for not only improving the quality of the skin graft but also extending the preservation time of the skin graft up to 14 days. PMID- 8719312 TI - Timing of the thermographic assessment of burns. AB - The thermographic assessment of burns using infrared imaging has previously been shown to be a useful aid in the estimation of burn depth. In this study, thermographic images of burns, obtained from 65 patients over a 4-year period, were reviewed. An infrared transparent, water-impermeable membrane was used as a wound cover to abolish evaporative cooling artefacts. Single images were obtained from patients with burns to various parts of the body, excluding the hands. A significant change in the temperature of deep burns was observed between days 2 and 3 after injury (chi-square, P < 0.01; Fisher exact probability test between days 2 and 3, P < 0.01). The results of this study suggest that thermography of burns, to assess depth, should be performed within 3 days following the injury. PMID- 8719313 TI - Epidemiological survey of childhood burn injuries in France. AB - Among domestic injuries in children, skin burns deserve special attention because they can lead to severe residual lesions. Their frequency has been reported to range from 3 to 8 per cent of all childhood injuries, but few epidemiological data have been collected in France. This study was undertaken to improve our knowledge of this public-health problem. A 20-item epidemiological questionnaire developed by the SFETB was used to collect prospective data on childhood burn injuries managed over a 12-month period in 14 burn centres and 18 paediatric surgery units in France. Of the 937 patients included in the study, 606 were contributed by burn centres and 331 by paediatric surgery units. Descriptive analysis and tests for correlations between several parameters showed that the typical burn-injury patient was a boy (61.6 per cent) with a mean age of 47 months, who suffered a scald burn (64.1 per cent) in the kitchen (56.2 per cent). The bathroom (13.6 per cent) was found to be an especially dangerous room because of the presence of hot water taps (shower, bathtub) and the high risk of an immersion injury. Burns due to ignition of volatile substances (16.95 per cent) were less common, but caused seven of the 11 deaths recorded in this survey. Most burn injuries were superficial lesions (59 per cent) covering less than 10 per cent of the body surface (56.2 per cent). However, grafting was required in 35 per cent of patients. Epidemiological surveys provide objective information on hazardous agents and on the settings in which burns are most likely to occur. They are essential for the development of appropriate preventive strategies, which are still receiving insufficient emphasis in the European Economic Community (EEC). PMID- 8719314 TI - Human keratinocyte isolation and cell culture: a survey of current practices in the UK. AB - A survey was conducted to establish current techniques for isolation and culture of human keratinocytes. A questionnaire was sent to all units thought to be involved in keratinocyte culture, a total of 34 individuals; 62 per cent of those surveyed responded to the questionnaires. The proportion of individuals using high-calcium medium to culture keratinocytes was 53 per cent, while 47 per cent used low-calcium serum-free medium. The majority of replies followed trends dependent on the culture method employed. Details of anatomical donor skin site, keratinocyte isolation and culture were compared. In particular, the problems associated with the use of commercially prepared low-calcium, serum-free medium were reported. The basic principles of keratinocyte culture reported in the literature were seen amongst all the replies received. It is interesting to note the variations in methods adopted as techniques are passed on and continuously modified to suit the requirements of the individual worker. This survey also highlights the difficulties that can occur when using mass-produced complex media. PMID- 8719315 TI - The use of calcium alginate dressings in deep hand burns. AB - Deep burns of the hand are a common serious surgical problem with major occupational and economic implications. Control of haemorrhage during excision and grafting is difficult and postoperative haematoma may reduce graft take. Following excision, important deep structures such as extensor tendons or joints may be exposed. Such tissues do not take free skin grafts satisfactorily. We have found calcium alginate dressings can be of immense help in minimizing these technical problems. We describe our experience in one patient; three other patients have been treated with equal success. PMID- 8719316 TI - Vibrio cholerae non-O1 primary septicaemia following a large thermal burn. AB - A case of primary septicaemia with a Vibrio cholerae not agglutinable with O group 1 sera is reported in a burn patient. This appears to be the first reported case of this organism causing infection in a burn patient. The case is discussed, highlighting the difficulties encountered in treating this unexpected organism and the course of the infection in this patient. It is probable that the organism was obtained during first aid for the burn wound. PMID- 8719317 TI - Aeromonas bacteraemia in burn patients. AB - Human aeromonas infection is uncommon and is usually associated with immunosuppression, chronic disease or trauma in an aquatic setting. Burn injury may induce a state of immunosuppression, making the thermally injured patient a suitable host for aeromonas infection. We reviewed the experience of one burn centre with this pathogen. Retrospective examination of blood culture results from 8151 patients admitted between 1959 and 1994 disclosed eight patients with clinically relevant Aeromonas hydrophilia bacteraemia. Five were burned outside the USA. Aquatic exposure was known or suspected in only three cases. Five of the eight patients died. Aeromonas infection in burn patients is rare but may occur in the absence of aquatic exposure. PMID- 8719318 TI - Management strategy in purpura fulminans with multiple organ failure in children. AB - Purpura fulminans (PF), which describes the necrosis of soft tissue secondary to diffuse microvascular thrombosis induced by transient protein C deficiency associated with meningococcal sepsis, is unusual despite the approximately 15000 cases of bacterial meningitis which occur annually in the USA. PF has a reported mortality of 50 per cent secondary to multiple organ failure which commonly accompanies the syndrome and is associated with major long-term morbidity in those who survive. Children who develop multiple organ failure in association with purpura fulminans are difficult management problems and benefit from the unique surgical and critical care resources available in burn centres. We describe our recent experience with three such patients and suggest a management strategy, the key components of which include early excision and closure of deep wounds, aggressive critical care management and long-term follow-up should delayed epiphyseal growth occur. PMID- 8719319 TI - Unusual development of granulomas on the healing surface of burn wounds associated with MRSA infections. AB - Ten patients with a mean age of 14.5 years and partial skin thickness burns involving 3-5 per cent body surface areas developed rapidly proliferating tumour like growths on the surface of their healing wounds within 10-21 days of sustaining the injuries. The number of tumours on every patient was either single or multiple and each increased in size daily. The growths were unique in their fulminating-type fleshy mass, with a consistency varying from soft to firm, absence of purulent material or head, and extension up to the fibrous layer covering the musculatuve. Histopathology was suggestive of granulomatous rather than a suppurative nature of the lesions. Isolation of MRSA from the burn wounds of four cases on the first day of dressing and then from the surface of the tumours of all of them and the excised tissues, as well as from the environment of the dressing room, indicated its involvement in the causation of the growths through contaminations of wounds with a hospital endemic strain while handling or dressing. The organisms were resistant to most antibiotics except vancomycin and teicoplanin. The growths in four cases subsided within 72 h with daily dressing, using an injectable solution of either vancomycin or teicoplanin, while the rest required radical excision and immediate cover with split skin grafts and systemic administration of either of the antibiotics. The wounds healed over a period of 8 10 days. PMID- 8719320 TI - Pharmacodynamics of mivacurium in severely burned patients. AB - The pharmacodynamics of mivacurium, a new short-acting non-depolarizing muscle relaxant, were studied in nine severely burned patients with concomitant inhalation injury. Complete neuromuscular blockade was achieved within 1.3 min (controls 3.0 min) following the usually recommended intubating dose (0.15 mg/kg/BW 2 x ED95) of mivacurium. The clinical duration of neuromuscular blockade and the recovery times were slightly prolonged, due to significantly reduced serum cholinesterase activity (clinical duration 24.6 min vs. 15.3 min). This pharmacodynamic profile makes mivacurium preferable for intermittent on-demand neuromuscular blockade in the severely burned patient. PMID- 8719321 TI - Three years of an adult burns support group: an analysis. AB - A retrospective analysis, by survey, of the benefits of a support group and preference for its structure is presented. Psychosocial support after burn injury is identified as important and a support group is recommended as being routine in the aftercare for burns' patients. PMID- 8719322 TI - Use of seven-flap plasty for the treatment of axillary and groin postburn contractures. AB - Axillary and groin contracture with limitation of abduction of the arm and leg are one of the most common complications of burns. In this paper we report the technique of using seven-flap plasty for treating axillary and groin scar contractures. The advantages of using this technique are that it does not need prolonged splinting and intense and painful physical therapy. PMID- 8719324 TI - Longitudinal trapezius fasciocutaneous flap for the treatment of mentosternal burn scar contractures. PMID- 8719323 TI - Acute pulmonary oedema following administration of vasopressin for control of massive GI tract haemorrhage in a major burn patient. AB - A 35-year-old male sustained a full-skin thickness chemical burn involving 60 per cent of TBSA when hydrochloric acid was applied to his face, trunk and extremities by his girlfriend. Debridements and skin graftings were performed smoothly and he was doing well until day 23 after injury, when massive GI tract bleeding caused a drop in blood pressure. Vasopressin was given intravenously to control the bleeding, which stopped, and the blood pressure returned to normal after transfusion. After the vasopressin infusion was tapered off acute pulmonary oedema developed abruptly, which required treatment by intubation and PEEP using a respirator. The lung condition had returned to normal by the following day. A second episode of massive GI tract bleeding recurred 10 days later, again vasopressin was given through a catheter into the inferior mesenteric artery. Again pulmonary oedema developed 38 h after the vasopressin use, the oedema disappeared within 2 days when the vasopressin infusion tapered off. It should be kept in mind that acute pulmonary oedema may develop when high doses of vasopressin are used in the treatment of Curling's ulcer or other GI tract bleeding. PMID- 8719325 TI - Ontogeny of oxytocin-like immunoreactivity in the Brazilian opossum brain. AB - The neuropeptide oxytocin (OT) has been shown to function as a neurotransmitter and/or neuromodulator in addition to its hormonal function in the periphery in the adult central nervous system (CNS). Previously, we have studied the postnatal neurogenesis of the paraventricular and supraoptic nuclei and ontogeny of arginine vasopressin-like immunoreactivity in the Brazilian opossum brain, Monodelphis domestica. In this study, we have described the ontogeny of oxytocin like immunoreactivity (OT-IR) in the opossum brain. As a marsupial, opossum pups are in an extremely immature state, with neurogenesis and morphogenesis continuing into the second week of postnatal life. Thus, opossum pups are a good model for developmental studies. In the adult opossum brain, OT-IR was localized in regions as reported for the adult rat and other species, except for a few differences. These findings suggest similar functional roles for OT in the adult opossum brain as in other mammals. Unlike the prenatal expression of arginine vasopressin, OT-IR was first detected in the forming median eminence on day 1 of postnatal life (1 PN). Between 3 and 5 PN, OT-IR was present in the hypothalamic supraoptic and paraventricular nuclei and posterior pituitary. At this time, neurogenesis of these nuclei is not completed. By 10 to 15 PN, OT-IR was seen in several brain areas, and begins to resemble that of the adult between 45 and 60 PN. These results indicate that the time course of appearance of the OTnergic system does not directly parallel the early expression of the vasopressinergic system. However, the expression of OT-IR in the opossum brain before neurogenesis and morphogenesis is completed suggests a potential role for OT in developmental events. Similar to arginine vasopressin, oxytocin may also be involved in the regulation of autonomic functions that are essential for the opossum's adaptation to an ex utero environment. Future studies utilizing experimental manipulations of the OTnergic system will help determine the significance of this peptide in the neonatal opossum. PMID- 8719326 TI - Phenotype plasticity and immunocytochemical evidence for ChAT and D beta H co localization in fetal pig superior cervical ganglion cells. AB - The early expression of the cholinergic phenotype in sympathetic neurons was already studied in superior cervical ganglion cells derived from rat, quail and chicken embryo. In the present work, we set up a neuron culture derived from the superior cervical ganglia of fetal pigs. The yield is 1000 times of that of a neonatal rat [17], 100 times of a 10- to 13-day-old chick embryo [26] and 20 times of a 10-day-old quail embryo [3]. This high yield will greatly facilitate further biochemical studies concerning neuronal differentiation. Using these cells as a model, the phenotype plasticity was studied by both biochemical and immunocytochemical methods in normal physiological medium, in a high KCl (30 mM) medium and in a splenocyte co-culture. The phenotype shift occurs in the normal physiological medium and in the splenocyte co-culture, but not in the high KCl medium. Taking into account the species difference, the fetal pig superior cervical ganglion neurons behave in a comparable manner as reported in earlier studies for other animal models. Moreover, for the first time, using immunocytochemical methods, direct evidence for a co-localization of choline acetyl-transferase and dopamine-beta-hydroxylase in mammalian fetal sympathetic neurons, at least during a certain period, is given. PMID- 8719327 TI - GAP-43 expression in the medulla of macaque monkeys: changes during postnatal development and the effects of early median nerve repair. AB - Expression of GAP-43, a neuronal specific growth associated phosphoprotein, has been highly correlated with the growth and remodeling of the nervous system during development and regeneration. As part of an effort to understand mechanisms of developmental plasticity in the somatosensory system, we determined how the expression of GAP-43 is affected by prenatal and early postnatal nerve cut and repair in macaque monkeys. We also observed normal developmental changes in the expression of GAP-43 during early postnatal life in macaque monkeys. The normal cuneate nucleus, as well as other nuclei of the ascending somatosensory pathways, had low levels of GAP-43 at birth that increased by 3 months and declined thereafter to reach adult levels between 8 and 15 months of age. Fiber tracts expressed low levels of GAP-43 at all postnatal ages, except the pyramidal tract which demonstrated high levels a birth that decreased over the first year. These observations suggest a gradual but differential synaptic maturation in lower brain stem nuclei as macaque monkeys mature. Greatly increased levels of GAP-43 were observed at the time of birth in the cuneate nucleus of two macaque monkeys with prenatal (E94 and El 14) nerve repair. Such an increase was not found after prenatal nerve repair with a postnatal survival time of 15 months, or after early postnatal nerve repair with short (80 days) or long (20 months) survivals. The results suggest that reorganization mechanisms at central terminals of peripheral nerves are very different following prenatal than postnatal nerve damage. PMID- 8719328 TI - Developmental injury to the cerebellum following perinatal Borna disease virus infection. AB - In rats infected as neonates, Borna disease virus (BDV) infection causes neuroanatomical, behavioral and physiological abnormalities without encephalitis. Neonatal infection with BDV provides a powerful model for studying the effects of virus replication on brain development without inflammation-induced brain damage. Here we report that neonatal BDV infection interfered with cerebellar development in the Lewis rat. Based on cerebellar cross-sectional area measurements, abnormal cerebellar growth was first noted between 7 and 14 days after infection. Reactive astrocytosis was evident by three days after infection, even without encephalitis, and even before identification of viral proteins in the cerebellum. While neonatal BDV infection caused a significant loss in granule cells, infected granule cells were not identified. BDV proteins were readily detected in the Purkinje cells. Thus, persistent BDV infection of Purkinje cells, but not granule cells, was associated with loss of granule cells during cerebellar development, in the absence of encephalitis. PMID- 8719329 TI - Postnatal changes in serine/threonine protein phosphatases and their association with the microtubules. AB - The activities and protein levels of three serine/threonine protein phosphatases were determined in homogenates and microtubule preparations from rat brain at various ages from postnatal day 1 (P1) through adulthood. The activities and levels of the calcium/calmodulin-dependent protein phosphatase, phosphatase 2B increased significantly from P1 to P21 in brain homogenates and remained elevated in the adult. The association of phosphatase 2B with microtubules was also found to be increased in the adult compared to the neonate (P3). In contrast, protein phosphatase 2A in brain homogenates decreased significantly from P1 to adult. However, the association of phosphatase 2A with the microtubules was found to increase with age. Finally, the activity and levels of phosphatase 1 in brain homogenates did not change with postnatal age, although the association of phosphatase 1 with microtubules was significantly decreased in the adult brain compared to P3. These studies clearly indicate that the activity, levels and association of these serine/threonine phosphatases with microtubules are independently regulated during postnatal development and suggest unique roles of phosphatase 1, phosphatase 2A and phosphatase 2B in modulating the phosphorylation state and function of microtubule-associated proteins at different postnatal ages. PMID- 8719330 TI - Few neocortial and thalamic morphological changes after a neonatal frontal cortical ablation contrast with the effects of a similar lesion in fetal cats. AB - To further understand the neuroanatomical consequences following perinatal brain injury, quantitative morphometric analysis was performed on the brain of cats receiving a unilateral frontal cortical ablation between postnatal days (P) 9 and P 14 and intact control cats. In all cats, the volume of the neocortex and thalamus was measured bilaterally and that of the thalamic ventrobasal complex (VBc) was measured ipsilaterally. In addition, using stereology, the neuronal and glial (presumably) cell packing densities (CPD) and the total number of neurons and glial cells (TCN) were measured in the ipsilateral VBc. The neuronal and glial cell cross-sectional areas (CSA) were also measured in the ipsilateral VBc. The mean ipsilateral and contralateral neocortex volumes were similar between the two animal groups. There was a statistically significant 14% and 13% reduction in mean ipsilateral and contralateral thalamic volumes, respectively for the lesioned animals, while the VBc shrank by 16% relative to intact controls. The mean neuronal and glial CPD were similar between the two groups. The mean neuronal TCN was reduced by 10% in the neonatal-lesioned cats, while the mean glial TCN was reduced by 31% in the same animals, however neither value reached significance. Lastly, the mean CSA of neurons and glial cells showed a tendency to be smaller in the lesioned cats by 8% and 9%, respectively. These results: (a) indicate that the neonatal lesion caused only minor morphological brain alterations and this sharply contrast with the marked changes previously reported in cats with a similar lesion sustained prenatally; (b) suggest that the enhanced behavioral recovery and/or sparing reported for the present cats compared to fetal-lesioned animals is at least partially due to the morphological sparing reported here; (c) together with previous findings in fetal cats, support the hypothesis that the morphological changes after a neonatal neocortical lesion are qualitatively different and may depend on different mechanisms as compared to those occurring after similar damage sustained prenatally. PMID- 8719331 TI - SCG10 expresses growth-associated manner in developing rat brain, but shows a different pattern to p19/stathmin or GAP-43. AB - The gene encoding SCG10 was originally isolated as a neuronal marker from neural crest derivatives, implying that this protein may contribute to fundamental neuronal properties. To examine the developmental change of SCG10 expression in brain, immunoblot analysis and in situ hybridization were performed in embryonic day 15 (E15), E19, postnatal day 0 (P0), P6, P14, P30 and P90 rat brains. The distribution of SCG10 mRNA was compared to those of its homologue, p19/stathmin, and the well-characterized growth-associated protein GAP-43. Overall expression of SCG10 in brain reached a peak at E19 and decreased gradually by P30 to the adult level. The expression pattern of SCG10 in E15 whole body was identical with that of GAP-43; both mRNAs were specifically detected in developing neuronal structures. p19/stathmin mRNA, on the other hand, showed widespread expression throughout the whole body. Expression patterns of the three mRNAs overlapped in many structures in the perinatal brain, yet each showed unique expression during postnatal development. For example, in the developing cerebellum, strong GAP-43 expression was found in the external granule cells, which are presumably extending parallel fibers, while SCG10 strongly hybridized in the internal granule cells which have reached their final position and begun dendrite outgrowth. The unique transient expression of p19/stathmin was found in the subventricular zone in the cortex, the white matter in the cerebellum, the optic nerve layer of the superior colliculus and the inner edge of the dentate granule layer in the hippocampus. Considering the timing, all of these areas are known to produce neurons or glia. This is consistent with the suggestion that p19/stathmin is related to differentiation. SCG10 may be a new member of growth-associated proteins and this protein may contribute to neurite extension in perinatal brain as does GAP-43. However, the differential expression between SCG10 and GAP-43 in later developmental stages suggests their diverse functions, which indicates these proteins may play different roles during postnatal brain development. PMID- 8719332 TI - Trophic and protective actions of brain-derived neurotrophic factor on striatal DARPP-32-containing neurons in vitro. AB - We have examined the effects of either brain-derived neurotrophic factor (BDNF), the BB-isoform of platelet-derived growth factor (PDGF-BB), or a combination of these growth factors on the survival and the morphological development of embryonic striatal neurons grown under serum-free culture conditions. Striatal neurons were identified using immunocytochemistry for "dopamine- and adenosine 3':5'-monophosphate-regulated phosphoprotein with a molecular weight of 32 kilodalton" (DARPP-32). BDNF and PDGF-BB promoted the survival of DARPP-32 positive neurons, with the magnitude of their effects being comparable. A combination of these growth factors exerted no significant additive effects on cell survival. BDNF stimulated morphological differentiation of DARPP-32 containing neurons by increasing the length of neurites, the number of branching points on the neurites, and the soma area. By contrast, PDGF-BB increased the neurite length and the cell body area, but not the number of branching points. BDNF also protected striatal neurons from excitotoxicity induced by N-methyl-D aspartate, whereas PDGF-BB had no effect under the same treatment conditions as those for BDNF. Thus, BDNF is trophic for striatal DARPP-32-containing neurons in vitro by enhancing the survival, morphological differentiation and resistance to excitotoxicity, and its mechanisms of action are probably different from those of PDGF-BB. PMID- 8719333 TI - Complex gangliosides affect GD3 accessibility to antibody in developing neuronal cells. AB - Ganglioside expression of embryonic chick retina cells developed in vitro was analyzed by indirect immunofluorescence. Immature neurons were GD3 positive cells and the labeling was chiefly distributed all over their cell membrane. Mature neurons became GD3 negative and expressed complex gangliosides of the a- and b pathways; nevertheless, the content of GD3 accounted for approximately 40% of the total gangliosides in these cells. Neuraminidase hydrolysis pointed out that GD3 was located in membrane of differentiated cells. The frequency of cells with the GD3 immunostain localized in restricted area of membrane of undifferentiated neurons increased significantly after adding a mixture of bovine brain gangliosides (largely complex gangliosides). Antibody binding to immobilized GD3 showed a dose-dependent inhibition by adding a mixture of bovine brain gangliosides, GM1, GD1a or asialo-GM1. Glycosphingolipids with shorter oligosaccharide chains, as cerebrosides or sulfatides, did not affect this binding. These results suggest that, concomitant with the accretion of content of complex gangliosides, a rearrangement in the membrane would occur, which progressively masks GD3 to its antibody. This rearrangement might affect putative ganglioside functions involved in neuronal differentiation. PMID- 8719334 TI - Branching pattern of corticothalamic projections from the somatosensory cortex during postnatal development in the rat. AB - In adult animals corticothalamic (CT) axons pass through the thalamic reticular nucleus (Rt) where they give off collateral branches innervating the Rt neurons. The postnatal development of CT projections from the somatosensory cortex, with particular reference to the branching pattern within Rt, ventrobasal (VB) and posterior (PO) nuclei, was investigated in the rat with anterograde tracing. Biotinylated dextran-amine (BDA) was iontophoretically injected into the somatosensory cortex of rats ranging from postnatal day (P) 0 to P30. At P1 most of the cortical axons traversed unbranched Rt and terminates in VB and PO, whereas at P3 they formed rudimentary branches in these nuclei. From P6 to P9 a progressive increase in the amount of dense clusters of terminal arborizations was evident in Rt, and by the second postnatal week more complex arborizations with a clear topographic arrangement were observed in Rt, VB and PO. Our findings indicate that CT fibers show a quantitative increase both in R1 and in somatosensory thalamic nuclei during the first postnatal week, although their terminal arborizations are however still incomplete. The pattern of collateralization of CT projections achieves an adult configuration at the end of the second postnatal week. PMID- 8719335 TI - K252a, a potent inhibitor of protein kinases, inhibits the migration of cerebellar granule cells in vitro. AB - In order to elucidate the cellular mechanisms of migrating neurons, we developed an assay system in vitro, using an aggregation culture of developing granule cells from the rat cerebellum. This assay system allowed us to eliminate the effects of various factors other than neurons and to examine the direct effects of individual molecules on neuronal migration. In this assay system, we examined the effects of several protein kinase inhibitors on cerebellar granule cell migration, and revealed that K252a, an inhibitor of protein kinases and of the actions of neurotrophins, inhibited the migration. Within 5 min after the addition of K252a to the culture medium, most of the migrating spindle-shaped cells changed into non-migrating large and polygonal cells, which had many microspikes. Staining with rhodamine-phalloidin revealed the appearance of actin bundles that resembled stress fibers within these large cells. On the other hand, extension of neurites was not severely inhibited by the addition of K252a. These results suggest that the migration is regulated by a different mechanism from that of neurite growth. PMID- 8719336 TI - Comparative localisation of CRYP alpha, a CAM-like tyrosine phosphatase, and NgCAM in the developing chick visual system. AB - The avian CRYP alpha gene is expressed in the embryonic nervous system and encodes a receptor-like protein tyrosine phosphatase with structural similarity to neural cell adhesion molecules. To gain further insight into the role of the CRYP alpha phosphatase in neural development, this study addresses the protein's cellular distribution in the well characterised embryonic visual system. High levels of CRYP alpha protein localise in retinal axons extending from the eye to the tectum throughout the major growth periods of these nerve processes. In addition, primitive inner plexiform layer processes in the retina, tectobulbar axons, and non-retinal fibres of the tectal stratum opticum, contain large amounts of CRYP alpha. Its presence in non-fasciculated processes suggests that CRYP alpha has a role other than in fasciculation in short range fibres. In contrast to CRYP alpha, NgCAM is confined largely to axon fascicles in the retina and tectum, consistent with its demonstrated role in fasciculation of cultured neurites. In cultured retinal neurons CRYP alpha proteins reside both in neurite processes and in growth cone membranes, implicating both of these as potential functional locations for the protein. Although CRYP alpha continues to be expressed in the later embryo, the strong, early expression suggests a significant developmental role in the initial growth or guidance of nerve processes. This applies both over the longer range in the retinotectal and tectobulbar projections and over the shorter range within plexiform layers. PMID- 8719337 TI - Neurons derived from P19 embryonal carcinoma cells develop responses to excitatory and inhibitory neurotransmitters. AB - Cells of the P19 line of embryonal carcinoma cells differentiate into neurons, astrocytes and oligodendrocytes following treatment with retinoic acid. The neurons from these differentiating P19 cultures synthesize a pattern of neurotransmitters that resembles that of neurons of the forebrain. We treated P19 cells with retinoic acid and then implanted them into the striatum of adult rats. After times ranging from 1 to 15 weeks post-implantation, brain slices containing the implanted tissue were prepared and used for intracellular recording of electrical activity and responsiveness to application of neurotransmitters. Within 2 weeks of implantation, the P19-derived neurons had developed responsiveness to the excitatory neurotransmitter glutamate and the inhibitory transmitters gamma-aminobutyric acid and glycine. These neurons also exhibited spontaneous synaptic potentials. The responses to glutamate appear to be mediated by N-methyl-D-aspartic acid as well as non-N-methyl-D-aspartic acid receptor subtypes. Gamma-aminobutyric acid evoked bicuculline-sensitive depolarizing responses in the younger grafts and biphasic depolarizing/hyperpolarizing responses in older ones. Responses to glycine were strychnine sensitive and also showed age-related changes from depolarizing to biphasic character. Synaptic potentials in the younger grafts were exclusively depolarizing, but in older ones both depolarizing and hyperpolarizing events were observed. The synaptic potentials appear to arise from synaptic connections between P19-derived neurons within the grafts. Many of the features of P19-derived neurons are similar to those of neurons in the developing forebrain. PMID- 8719338 TI - The entry of acidic amino acids into brain and CSF during development, using in situ perfusion in the rat. AB - Previous studies using the rapid single pass blood to tissue uptake of substances by the capillaries of the blood-brain barrier, have failed to show significant uptake of acidic amino acids. However, by the use of a bilateral in situ brain perfusion in neonatal and adult rats, extending the perfusion time to 30 min, the carrier-mediated uptake of aspartate and glutamate into brain and CSF has been demonstrated. The ratios of 14C-acidic amino acids in the brain and CSF to that in perfusate were measured and represented as Rbrain and RCSF respectively, after 30 min, neonatal (1-week-old) Rbrain values for both amino acids were approximately twice that of adults, while neonatal RCSF for aspartate and glutamate were 3 to 5 times that of the adult. In contrast, there was no significant entry of NMDA into either compartment for both adults and neonates. The transfer coefficient, Kin into brain and CSF was also measured in relation to stages of development. In general the Kin values for brain and CSF for aspartate and glutamate were higher in the younger age groups than the adult group (1 week > 2 week > 3 week > or = adult). In 1- and 2-week-old rats entry into CSF appears to be higher than that of brain, whereas for adults entry into the brain tissue was dominant. PMID- 8719339 TI - The role of the cholinergic system in the development of the human cerebellum. AB - High affinity (-)nicotine ([3H]nicotine), alpha-bungarotoxin ([125I]alpha bungarotoxin) and muscarinic binding ([3H]N-methyl scopolamine) in the human cerebellum were compared between the foetal period (23-39 weeks gestation) and young adulthood (14-34 years) in an autoradiographic study. To estimate proportions of muscarinic receptor subtypes variable wash times and displacement with pirenzepine were employed. [3H]Nicotine binding and total muscarinic binding in foetuses exceeded that in young adults by a factor of 6 and 2 respectively in the dentate nucleus, and by a factor of 3 in white matter. [3H]Nicotine and muscarinic binding was also higher in the foetal external granule cell layer than in the internal granule cell layer of adult, [125I]alpha-Bungarotoxin binding was raised in the dentate nucleus of the foetus compared with the adult. The M2 subtype appeared to be the predominant muscarinic receptor in the cerebellum, however it tended to represent a lower proportion of the muscarinic binding in the foetus than the adult. All 3 receptor types were highest in the foetal brainstem where the M3 + M4 muscarinic subtypes appeared to predominate. The p75 nerve growth factor receptor, measured by immunocytochemistry, in common with cholinergic receptors, paralleled choline acetyltransferase activity which has previously been reported to be high in the cerebellum during late foetal development and to fall in adulthood. PMID- 8719340 TI - Effects of neonatal and adult enucleation on the synaptic organization of the serotoninergic projection to the superficial gray layer of the hamster's superior colliculus. AB - Previous experiments from this laboratory have demonstrated that either neonatal or adult enucleation results in a marked increase in the density of the serotoninergic (5-HT) projection to the superficial layers of the hamster's superior colliculus (SC). The present study was undertaken to determine whether this alteration in the density of the 5-HT projections to SC in hamsters that sustained eye removals was associated with any change in the synaptic organization of this pathway. In normal adult hamsters, only 4.0% of 500 5-HT immunoreactive profiles made synaptic contacts in the stratum griseum superficiale (SGS). Of these, 65% were axoaxonic, 30% were axodendritic, and 5% were axosomatic. In contrast, 22% of 400 5-HT-positive profiles in the neonatally enucleated hamsters and 17.5% of 400 such profiles in hamsters that sustained eye removals in adulthood made synaptic contacts. In the neonatal enucleates, 66% of the contacts were axoaxonic, 33% were axodendritic, and 1% were axosomatic. In the adult enucleates, 73% of the contacts were axoaxonic, 26% were axodendritic, and 1% were axosomatic. Thus, both neonatal and adult enucleation increase the percentage of 5-HT-positive profiles that make synaptic contacts in SC, but neither manipulation significantly changes the distribution of these contacts among different types of postsynaptic targets. PMID- 8719341 TI - Developmental expression of the immediate early gene EGR-1 mirrors the critical period in cat visual cortex. AB - Immediate early gene (IEG) expression in the central nervous system is thought to play a role in coupling extracellular stimulation with the transcriptional events responsible for long-term functional changes in neurons. The goal of the present study was to determine the postnatal developmental profile of EGR-1 protein (also termed zif268, Krox-24, NGFI-A) expression across the layers of cal visual cortex and relate it to the state of visual cortical development and plasticity. Using a polyclonal antibody, EGR-1 immunoreactivity was studied in animals of various postnatal ages (from 0.5 week to adult). In very young animals (0.5 weeks), EGR-1 positive cells were restricted to deep cortical layers (layer VI/Subplate). With the increasing age, EGR-1 immunoreactivity spread across layers of the visual cortex in an inside-outside manner, and by 5 weeks of age, EGR-1 protein was highly expressed in all layers. EGR-1 expression remained high until approximately 10 weeks of age and then gradually began to decline in layer IV with little change in supra- and infragranular layers. In adult animals, EGR-1 was located predominantly in the layers above and below layer IV. This pattern of EGR-1 expression in developing cat visual cortex has both temporal and laminar similarities with the development of visual cortical connectivity, with the development of orientation selective receptive field properties, and with the level of visual cortical plasticity, suggesting an involvement of EGR-1 expression in these processes. PMID- 8719342 TI - Ontogenic development of dopamine D4 receptor in rat brain. AB - The postnatal development of rat brain dopamine D4 receptor gene expression was studied in animals 1 day to 1 year old, using the polymerase chain reaction technique. The level of expression of the D4 mRNA was appreciable at birth (day 1), increased to maximum at day 3, and showed declines at day 28. D4 mRNA expression remained unchanged at the ages of 6 months to 1 year. The mRNA expression at day 1 is about 50% of that observed on day 3, and declines approximately 50% by day 28. In contrast, the dopamine D2 mRNA expression was maximum at day 28, which is consistent with the reported studies. PMID- 8719343 TI - Mash-1 is expressed during ROD photoreceptor differentiation and binds an E-box, E(opsin)-1 in the rat opsin gene. AB - It has been shown previously that the transcription factor Mash-1 plays a significant role in mammalian neurogenesis. In the present study we show that Mash-1 is expressed during rat retinal development and that it interacts specifically with an E-box identified in the promoter of the opsin gene during rod photoreceptor differentiation. We propose that opsin may be one of the downstream neuronal genes regulated by Mash-1 during neurogenesis in the central nervous system. PMID- 8719344 TI - Regulation by nerve growth factor of neuropeptide phenotypes in primary cultured sensory neurons prepared from aged as well as adult mice. AB - The present study investigated neuropeptide phenotypes of aged, as well as adult, mouse sensory neurons. Proportions of somatostatin (SOM), calcitonin gene related protein (CGRP) and neuropeptide Y (NPY) immunoreactive (ir)-neurons were lower in primary cultures from aged (2 years) mice than in those from adult (6 months) animals, but similar for substance P (SP) in the absence of exogenous nerve growth factor (NGF). Addition of NGF, significantly enhanced (P < 0.05) proportions of SP, NPY and CGRP ir-neurons in both adult and aged cultures, whereas SOM ir-neurons were not affected in either. Thus SP, CGRP, NPY and SOM phenotypes are retained in cultured aged DRG neurons and some phenotypes can remain sensitive to NGF regulation. PMID- 8719345 TI - Evidence for the elaboration of multiple axons by developing dentate granule cells. AB - A sample of 185 Neurobiotin-filled dentate granule cells has been collected from rats aged P14 to P120 in a study of the effects of serotonin (5 hydroxytryptamine, 5-HT) depletion on granule cell development. A small number (5.9%) of these neurons exhibit more than one axon. These neurons have morphologies consistent with that described for dentate granule cells. One axon typically arises from the soma while additional axons take origin from either dendrites or the soma. Both axons may be mossy fibers or one axon may be a mossy fiber and the second assume a morphology and distribution similar to that described for interneurons. These latter neurons therefore exhibit a mixed phenotype by having a granule cell morphology and an interneuronal axon type. These data suggest that some granule cells give rise to multiple mossy fibers or express a mixed axonal phenotype during maturation. PMID- 8719346 TI - Developmental regulation of various annexins in the embryonic palate of the mouse: dexamethasone affects expression of annexin-1. AB - The annexins are a group of structurally related proteins implicated in a number of cellular processes, including growth, membrane fusion, and the effects of glucocorticoids on cellular physiology, signal transduction, and regulation of activities of phospholipase A2. Though their exact role in cellular physiology is not clear, their properties make them candidate proteins for signal transduction pathways by which growth factors and glucocorticoids modulate development of the palate. We have determined the exact cellular location and development expression of various annexins in the embryonic murine palate as a first step in assessing their developmental function. Western blot analysis revealed an increased accumulation of selected annexins in elevated palates compared to vertical (unelevated) ones. This was particularly striking for lipocortin I1 (annexin I), whose mRNA accumulated as well. Lipocortin I was expressed primarily in the apical portion of the palatal epithelium at early stages of development, but throughout the epithelium at later stages. Also, there was increased immunoreactivity for lipocortin I in the mesenchyme as development proceeded. Immunoreactivity for the endonexins (annexins IV and V) was found in the palatal epithelium and mesenchyme, whereas immunoreactivity for the 67-kDa calelectrin (annexin VI) was found only in the mesenchyme. Treatment of pregnant A/J strain mice with a cleft-palate inducing regimen of dexamethasone stimulated accumulation of lipocortin I protein and mRNA, but not lipocortin II (annexin II) protein. In contrast, the same regimen of dexamethasone did not affect levels of lipocortin I protein in palates of the glucocorticoid-less sensitive C57BL/6J strain mouse embryo. These data permit the suggestion that lipocortin I plays some critical, but as yet undefined, role in modulating ontogeny of the murine palate. PMID- 8719347 TI - Alterations in cranial morphogenesis in the Lp mutant mouse. AB - The effects of exencephaly on cranial morphogenesis were studied at 10 to 12 days of gestation in the loop-tail (Lp) mutant mouse in which the hindbrain and spinal cord fail to close. At the level of the hindbrain, the otocysts became displaced ventrally in abnormal (Lp/Lp) embryos, and the everted neuroepithelium showed a diminished "luminal" reaction to the lectins WGA and Con A, as compared with normal embryos. Also, occasional clusters of rounded cells that resembled presumptive neural crest cells and that labeled with WGA, Con A, and anti-N-CAM were observed at the everted tips of the open neural folds. By 12 days' gestation, there was a loss of integrity in some areas of the neuroepithelium. However, despite the topographic and neuroepithelial distortions, normal differentiation of a roof plate-like neuroepithelium occurred at the ends of the everted neural folds. In addition, the mesenchyme showed normal condensations that labeled with WGA, Con A, and anti-N-CAM in the perinotochordal basicranium and periotic regions. Thus, in this mutant mouse model of neural dysraphism, some features of cranial morphogenesis and differentiation appear to be dependent on timely and proper closure of the cranial neural tube, whereas other aspects may proceed independently of neural closure. PMID- 8719348 TI - Locally released retinoic acid leads to facial clefts in the chick embryo but does not alter the expression of receptors for fibroblast growth factor. AB - Systemic administration of retinoic acid (RA) affects the growth of the upper beak of chick embryos; however, the mechanism for generating a cleft upper beak is not known. In the present study, we wished to elucidate the molecular basis of the retinoid-induced lip clefting. In order to ensure that facial prominences were locally exposed to levels of retinoid known to affect gene expression, we implanted beads soaked in different concentrations of RA in the right nasal pit or in the centre of the frontonasal mass. Beads soaked in 5 mg/ml RA placed in the right nasal pit caused full clefting of the upper beak with a deviation of the midline toward the right side of the face. The asymmetry was principally due to a decrease in size or total elimination of the right lateral nasal prominence. RA-soaked beads placed in the centre of the frontonasal mass created full bilateral clefts that were more symmetrical than those produced by beads in the nasal pit. Lower concentrations of retinoic acid produced less severe facial abnormalities. Control experiments show that the implanted bead itself has no effect on growth or fusion of the facial prominences. The specific effects of retinoids on facial growth may be due to a localized decrease in responsiveness to growth factors. Gene expression patterns for two fibroblast growth factor receptors (Cek-2, Cek-3, [chicken embryo kinase]) in normal and RA-treated embryos were examined by in situ hybridization. In normal embryos, Cek-2 and Cek 3 transcripts are expressed at very high levels in the mesenchyme directly adjacent to the eye. Cek-3 is additionally expressed in the centre of the frontonasal mass. The application of beads to the right nasal pit did not change the level of expression or distribution of transcripts for Cek-2 or Cek-3. This data suggests that retinoic acid may be affecting other aspects of the FGF receptor-ligand interaction. PMID- 8719349 TI - Growth in the lateral part of the human skull during the second trimester. AB - The aim of the present study was to report dimensional changes in the lateral part of the prenatal human skull during the second trimester. The sample comprised 37 fetuses obtained in connection with requested autopsy. Gestational ages (GA) were available for 27 fetuses, ranging from 13 to 24 weeks. Crown-rump lengths (CRL) of 23 fetuses ranged from 78 to 228 mm. For 21 fetuses both CRL and GA were available. After excision of the midsagittal segment of the skulls by two paramedian sections from the frontal bone to the foramen magnum through the optic foramina, an incision was made through the middle of the occipital squama, bisecting the skull. The present study was based on the skull components of the resulting 74 lateral cranial tissue blocks. The tissue blocks were radiographed with a Hewlett Packard Faxitron unit. Seven reference points were digitized from each film and eight linear dimensions were calculated. Method errors ranged from 0.12 mm to 0.52 mm. The dimensions were related to crown-rump length (CRL) and lateral segment developmental stages (LSS). The increases in the linear dimensions of the anterior part of the lateral skull segments were similar to those found previously for the median skull segment. The dimensions of the anterior part of the lateral skull segments were found to increase by 150% during the second trimester, while those of the posterior part increased by 100% during the same period. Between maturity stages LSS 4 and 5 the dimensions increased by 30-50%, whereas less increase was observed between stages 5, 6, 7, and 8. This reduction was attributed to the progressive ossification of the cranial base. The almost horizontal distance from the mandibular condyle (condylion) to the anterior tympanic ring did not change significantly during the second trimester. Comparison of the bilateral skull segments showed no systematic asymmetry, but a significant individual biological variability in symmetry with standard deviations of 0.7-1.8 mm was found. PMID- 8719350 TI - Histological observations of teeth and peridental tissues in cleidocranial dysplasia imply increased activity of odontogenic epithelium and abnormal bone remodeling. AB - Cleidocranial dysplasia (CCD) is a heritable generalized bone dysplasia presenting with a variety of dental abnormalities. To delineate morphological features of the dental tissues, we have analyzed histologically the structure of teeth and peridental tissues obtained from four CCD patients. Dentin appeared regular, except when formed in response to caries, physiological attrition or root resorption. Both acellular and cellular cementum were present in deciduous teeth, whereas in permanent teeth, cellular cementum was virtually lacking and acellular cementum was partially hyperplastic. Enamel pearls were seen in the furcations of one deciduous and one permanent molar. The roots of most deciduous teeth were resorbed only to a slight extent. Morphologically, supernumerary teeth resembled their normal counterparts. Denticles as well as epithelial cell clusters were occasionally present in the root pulps of deciduous teeth. In the peridental tissues, epithelium was locally abundant, and the cytokeratin profile confirmed its odontogenic origin. While the organization pattern of the alveolar bone matrix related to unshed deciduous teeth and unerupted permanent teeth varied, woven bone was abundant. Both formative and resorption surfaces were seen. Also, reversal lines were prominent, suggesting that the bone had, in fact, undergone remodeling. While no direct association between the aberrant bone morphology and the multifarious histological abnormalities of dental tissues can be established, the local abundance of odontogenic epithelium in peridental tissues of developing/unerupted as well as fully developed teeth may be causally related to the formation of excess acellular cementum, enamel pearls and supernumerary teeth. PMID- 8719351 TI - Human fetal pituitary gland in holoprosencephaly and anencephaly. AB - The normal prenatal development of the human pituitary gland and the gland supporting sella turcica has recently been investigated. The sella turcica area constitutes a developmental boundary area in the cranial base. Posterior to the area the cranial base has developed close to the notochord, and anterior to the region the cranial base development is dependent chiefly on neural crest cell migrations. In the present study the sella turcica region was analyzed in two fetuses with holoprosencephaly (cyclopia and median cleft) and four fetuses with anencephaly combined with rachischisis in the neck region (GA 16-20 weeks). The sella turcica region was investigated radiologically and histologically. Adenohypohyseal gland tissue was localized by immunohistochemical hormonal marking. In both types of malformation an open craniopharyngeal canal was seen in the base of the sella turcica with adenohypophyseal glandular tissue located in the sella turcica, in the canal, and in the pharyngeal connective tissue at the external side of the cranial base. In conclusion, severe malformations of the pituitary gland occur in both holoprosencephaly, which is a polytopic field defect located anterior to the sella turcica, and in anencephaly associated with notochordal insufficiency posterior to the sella turcica. This might indicate that the sella turcica area, bounding different developmental fields, is involved in various craniofacial malformations. It is consequently recommended that examination of the pituitary gland should become a part of the routine autopsy of prenatal material when malformations in the face, brain, and cranial base occur. PMID- 8719352 TI - Aspirin, a silent risk factor in urology. AB - Aspirin is a widely used drug for its analgetic, antiinflamatory and antipyretic effects as well as for prophylactic effect in cardiovascular diseases. However, an increased number of operative hemorrhagic complications in patients on daily aspirin have been reported, an adverse effect highly relevant in urology. In this review the normal hemostatic mechanism and the chief pharmacological effect of aspirin on hemostasis is described. The literature is reviewed for hemorrhagic complications to aspirin in urology. Few reports indicate that aspirin increases bleeding and need for transfusion following prostatectomy, but no placebo controlled clinical trials with large patient groups have been carried out. Following prostate biopsy and extracorporeal shock wave lithotripsy aspirin induced hemorrhagic complications have been reported. Cessation of aspirin ingestion one week prior to invasive urologic procedures and correction of bleeding complications with desmopressin, platelet concentration or fresh whole blood is described. PMID- 8719353 TI - Extended measurement of glomerular filtration rate and effective renal plasma flow in ambulatory patients. AB - We describe a standardized clearance method over 5 h (one hour equilibration followed by eight consecutive 30 min clearance periods [period 2-9]) for the estimation of GFR (iothalamate I125) and ERPF (hippuran I131) during water diuresis in ambulatory and exercising patients. Four groups were examined. In group I (normal controls, n = 15) there were no significant changes in GFR, ERPF and FF (P > 0.10) during repeated clearance periods (mean of period 2-5 versus period 6-9). The reproducibility of the method was studied at a mean interval of 3.7 weeks in a group of patients with stable reduction of GFR (group II, n = 7). The values for GFR, ERPF, FF and RVR did not change significantly in this group, and correlated significantly between repeated studies (r = 0.81 to r = 0.99). In group III (untreated hypertensive patients with reduced GFR, n = 13) there was a time dependent 7.2% decrease in GFR (P < 0.05), significantly different from group I (P < 0.02), a 10.0% decrease in ERPF (P < 0.01) and no significant change in FF (P = 0.08) when the mean of period 2-5 was compared with the mean of period 6-9. In healthy controls (group IV, n = 8) light sustained bicycle exercise (25 W) induced a 7.1% decline in GFR (P < 0.01), 17.4% decline in ERPF (P < 0.001) and a 13.6% increase in FF (P < 0.001). We conclude that ambulatory measurements of GFR and RPF can be carried out over a period of 5 h with satisfactory precision and repeatability. Ambulatory hypertensive patients with moderately reduced GFR showed the same degree of time dependent downward drift of GFR and ERPF without exercise as was seen in healthy individuals during light exercise. Accordingly, in these groups single clearance periods imply a risk for under or overestimation of renal function, and time controls are necessary during clearance studies. PMID- 8719354 TI - Biochemical markers of renal disease in primary Sjogren's syndrome. AB - Primary Sjogren's syndrome (SS) is characterized by an inflammatory process in the salivary and lacrimal glands, but the kidneys may also be involved. Renal tubular functions were studied in 27 patients with SS, all females, age 37-78. Both SS-patients with and without known distal renal tubular acidosis (dRTA) were included, dRTA was found in 18/27 (67%), impaired urine concentrating ability in 13/27 (48%). Hypocitraturia was identified in 20/27 (74%) and reduced tubular reabsorption of phosphate (TRP%) in 18/27 (67%). Tubular proteinuria (alpha 1 mikroglobulin) was present in 11/24 (46%), and tubular enzymuria (NAG) in 7/24 (29%). Hypocitraturia and/or dRTA were found in all patients with any kind of abnormal renal tubular function test. All except one of the patients with dRTA not treated with sodium bicarbonate had hypocitraturia. We conclude that distal tubular dysfunction was common in our SS-patients, but a concommitant proximal dysfunction was also seen. Determination of urinary citrate represents a valuable test for detection of renal disease in SS. PMID- 8719355 TI - Original renal disease in a kidney-transplant population. AB - Classification of the underlying renal disorder in kidney transplant patients involves some uncertainties. To allow evaluation of the risk of recurrence of renal disease in the transplanted kidney and other characteristics and risks inherent to the specific renal disorder we have investigated the basis for and reevaluated the diagnosis of 1000 consecutive patients who received transplants in Goteborg 1985-1993. In the original registry, 36% of patients had been given the diagnosis chronic glomerulonephritis but the diagnosis was confirmed by biopsy in only half of them, 18% of the total population. Systemic vasculitides and hereditary disorders other than adult polycystic kidney disease which constituted 3% and 6%, respectively, had frequently been overlooked. The term chronic pyelonephritis (15%) covered a variety of conditions including toxic tubulointerstitial disease, but was dominated by those caused by congenital urinary tract formations. Diabetic nephropathy (21%) was the consequence of Type 1 diabetes in 18% and Type 2 or other forms of diabetes in 3%. The proportion of patients with unknown cause of renal failure was 20%. The registry allows identification of small, distinct entities, which may be characterised as regards prerequisites for and consequences of kidney transplantation. PMID- 8719356 TI - Correction of post-renal transplant erythrocytosis by enalapril. AB - We studied whether post-renal transplant erythrocytosis (PRTE) could be corrected by enalapril with minimal side-effects, thus avoiding iterative phlebotomies or bilateral nephrectomy of native kidneys. From our renal transplant patients, 12 presented a true PRTE as defined by a 51-Cr red blood cell mass (RBCM) above 32 ml/kg for women and above 36 ml/kg for men. Secondary polycythemia was ruled out: all the patients had a normal renal artery pulsed ultrasonography; in all cases the blood arterial 02 saturation was above 96%. Bone marrow aspiration and histology were performed for each patient: none of them showed evidence of Vaquez disease. All of them had stable renal function i.e. the mean serum creatinine was 112.8 +/- 26.3 mumol/l. They all received the same immunosuppression: azathioprine; ciclosporine A; methylprednisolone. PRTE occurred within the first year post transplant (median 7.5 months; range: 2-34). Their mean RBCM was 37.38 +/- 2.7 ml/kg. Their mean serum value of Epo was 17.41 +/- 13.5 mU/ml (range: 9.1 54). After informed consent, all patients received enalapril starting with 5 mg/day, progressively increased to 20 mg/day, if necessary, in order to maintain the hematocrit below 45%. The mean daily dosage of enalapril was 13.75 +/- 6.1 mg (range: 5-20). The mean follow-up was 14.8 months (range: 3.5-29.5). There was no change in renal function (mean serum creatinine: 126.3 +/- 35 mumol/l). A successful response to enalapril was obtained with a median of 40 days (range: 20 120). 11 patients out of 12 responded to enalapril with a decrease of Hb (14 +/- 2 g/dl vs 16.8 +/- 1.04 g/dl; p = 0.0006) and Ht (41.9 +/- 6.17% vs 51.14 +/- 2%; p = 0.0002) without a significant decrease of Epo (8.1 +/- 3.87; p = 0.1). One patient did not respond to enalapril nor to captopril, but did respond to a combined treatment of enalapril and theophilline. Moreover, all PRTE patients but two did not have Epo levels, before enalapril, above the normal range, suggesting mechanisms other than Epo overproduction by native kidneys i.e. erythropoiesis dysregulation. In conclusion, all patients but one were successfully treated by enalapril without side effects. The treatment was effective as early as 3 weeks from the start and avoided the need for iterative phlebotomies and nephrectomy of native kidneys. PMID- 8719358 TI - Study on calcium oxalate monohydrate renal uroliths. I. Qualitative properties. AB - Shape, colour, surface features and external appearance were determined from 33 human uroliths composed predominantly of calcium oxalate monohydrate (COM). Based on these properties COM renal stones were classified into mulberry (M) and spheroid (S) type, each of which was further divided into two well defined subtypes, M1-fused globules, M2-loose globules, S1-corrugated surface and S2-even surface. This classification indicates that only specific combinations of external characteristics could occur on COM renal uroliths. No apparent correlation between the stone type and respective biochemical urinary data transpired from available information. PMID- 8719357 TI - Renal tubular dysfunction after urinary diversion. AB - Renal tubular function can be assessed by measuring low molecular weight proteins in urine. In the present study, urinary levels of protein HC, also called alpha 1 microglobulin, were determined in 84 patients, 3-17 years after conduit diversion or continent urinary reconstruction. Increased excretion of protein HC, indicating tubular dysfunction, was found in 30 patients (36%), but was slight in most cases. Among patients with GFR > 45 ml/min/1.73 m2 at latest follow-up, the fall from preoperative GFR was greater in those with elevated levels of protein HC than in those with normal levels (p < 0.01). No permeation of protein HC through the intestinal mucosa in contact with urine could be demonstrated. Urinary protein HC may be a suitable marker for detecting early renal impairment after urinary diversion. PMID- 8719359 TI - A study on calcium oxalate monohydrate renal uroliths. II. Fine inner structure. AB - The inner fine structure of 30 human uroliths composed predominantly of calcium oxalate monohydrate was studied in detail. Each type of stone distinguished on the basis of qualitative parameters, viz. M1, M2, S1 and S2 (see Part I), exhibited a specific and characteristic inner structure different in several well defined aspects from the other types. The inner structure suggests a sedimentary origin of the M1 type stone whereas the fixed particle origin of the M2, S1 and S2 stones, 3 types of core on which M2, S1 and S2 calculi developed were identified. The A type was represented by a void cavity with walls covered by an organic matter, the B type was formed by loosely arranged COM crystals and the C type was represented by a layer of an organic matter. Clinical observations lend support to the sedimentary origin of the M1 stones. PMID- 8719360 TI - Study on calcium oxalate monohydrate renal uroliths. III. Composition and density. AB - Density and content of mineral constituents were determined for 33 human calcium oxalate monohydrate (hereafter COM) uroliths whose external appearance and inner structure were described in part I and II respectively. Studied stones contained 0.13-0.42 wt.% of struvite, 0.68-4.12 wt.% of hydroxyapatite, 73-96 wt.% of COM and 3-10 wt.% of water unbound in a crystallohydrate 10 to 20 wt.% of calculus mass is not accounted for by chemical analysis. Density of COM calculi varying between 1.67 and 2.06 g cm-3 is not a function of any single stone parameter. Around 30% of stone volume is not occupied by crystalline components. The mulberry stones of sedimentary origin contained higher amount of organic matter than papillar and mulberry stones displaying site of attachment to epithelium. PMID- 8719362 TI - Burch colposuspension for stress urinary incontinence. 5-year results in 153 women. AB - The long-term results of Burch colposuspension for stress urinary incontinence were evaluated in 153 women operated on in 1984-1990. At follow-up 2 months postoperatively, 88% of the women reported continence, 5% were almost continent and the failure rate was 7%. At 24 months the corresponding figures were 86.7 and 7% and 39-102 months (mean 5 years) postoperatively they were 78, 11 and 11%. There were few complications. Follow-up urodynamic investigations showed significant rise of the pressure transmission from bladder to urethra in the cured patients. Reasons for failure are discussed. For patients with low urethral closure pressure, a vaginal sling procedure is preferable to Burch operation. Burch colposuspension is safe and advisable for patients with genuine stress incontinence. Urodynamic evaluation thus should be made preoperatively, but need not to be repeated if the clinical result is good. PMID- 8719361 TI - Cost and effectiveness of different treatment alternatives in urinary stone practice. AB - The cost and effectiveness of seventy-six consecutive percutaneous nephrolithotomy (PNL) procedures performed during the years 1990-1992, a sample of 425 ESWL patients with 675 treatments from 1991-1992, and 45 successive open pyelolithotomies (PL) performed before the advent of the new stone treatment techniques during the years 1981-1985, were studied for each of the treatment modalities. The costs were counted per patient and based on the cumulative charge of the procedures, cost for in-patient care, and treatment of additional procedures and operatively treated complications. The results showed that the great majority of ESWL patients were treated at a low cost and with few additional procedures and complications compared to PNL, particularly, when stones smaller than 20 mm were treated. Considering all the patients, the cumulative mean hospital cost per patient without the cost for the follow-up was as follows, FIM (pound): PL 33860(4200), PNL 63360(7860), and ESWL 17430(2160). The remarkable number of additional and auxiliary measures (including the pre- and post-operative ESWL procedures) resulted in considerable extra costs for the PNL patients until they were rendered stone-free. Another series comprising 42 successive patients with 48 stone basket procedures (SB) from the years 1981-1985 was retrospectively compared to 42 patients with 50 ureteroscopic stone extractions (URS) between January 1985 and April 1988 before the beginning of the ESWL practice in Finland as well as to 54 patients with 79 ESWL treatments for a lower ureteric stone during the years 1991-1992. The stone-free status after the three procedures until one month was 71, 66, and 60%, with no statistically significant difference. In the group of lower ureteric stones the cumulative mean hospital cost per patient was as follows, FIM (pound): SB 19520(2420), URS 17750(2200), and ESWL 17810(2210). Also in the treatment of lower ureteric stones, ESWL is an equally cost-effective method representing an alternative worth consideration, particularly for patients preferring a minimally invasive urinary stone treatment. PMID- 8719363 TI - The ice-water test in the diagnosis of detrusor-external sphincter dyssynergia. AB - The ice-water test (IWT) implies rapid intravesical infusion of 100 ml of sterile ice-water during continuous pressure measurement and registration of fluid leakage. In a typical positive test, there is fluid leakage around the catheter(s) during the peak of detrusor contraction elicited by cold stimulation. Seventy-six patients, the majority with different forms of spinal disorders and a clinical and urodynamically suspected detrusor-external sphincter dyssynergia, were subjected to cystometry, needle electromyography (EMG) and an ice-water test. Detrusor-external sphincter dyssynergia was found in 44 (59%) patients and 41 of them had a positive IWT. A positive test with a high detrusor pressure indicates detrusor-external sphincter dyssynergia whereas the contrary applies to the negative test. Eighteen patients who responded to cold stimulation with detrusor contraction but without fluid leakage, called positive non-leakage IWT, all presented detrusor-external sphincter dyssynergia according to EMG. In this situation, the cheap, non-invasive and simple IWT can replace a needle EMG study. PMID- 8719364 TI - Orthotopic bladder substitutes: histopathologic risk factors. AB - Prostate glands from 150 patients with carcinoma of the bilharzial bladder who underwent cystoprostatectomy were studied histopathologically by step sections. Prostatic urethral involvement by urothelial carcinoma was noted in 13 out of 96 (13.5%) and 5 out of 40 (12.5%) squamous and transitional cell tumors, respectively. None of the 12 adenocarcinomas and the two undifferentiated tumors showed involvement. Prostatic urethral involvement was as high as 19% in basal tumors and 26.7% in multifocal tumors compared to only 6.5% when the tumors occupied the bladder body. There was a significant increase in the incidence of prostatic urethral involvement from 9.5 to 35% when the prostate gland was involved. Prostate gland was involved in 20 out of 150 (13.3%). The bladder tumor was basal and infiltrating the prostate in 18 such cases. Seminal vesicles were infiltrated in 6 cases from the adjacent basal bladder tumors. We conclude that patients with basal or multifocal tumors are risky regarding bladder substitution and we recommend routine diagnostic transurethral prostatic biopsies and frozen sections from the site of urethral transection during cystoprostatectomy whenever bladder substitution controlled by the urethral sphincter is considered. PMID- 8719365 TI - Does anamnestic symptom evaluation or clinical examination give enough information to evaluate the severity of obstruction in benign prostatic hyperplasia? AB - In this study we have investigated 70 men fulfilling the usual criteria accepted for transurethral resection of the prostate (TURP). The anamnestic evaluation included the Madsen-Iversen symptom score and a quality of life questionnaire. The clinical examination included suprapubic pressure flow measurement, free urinary flow, the determination of residual urine and the ultrasound evaluation of the size of the prostate. The clinical data were correlated with the grade of obstruction according to Schafer calculated from the pressure/flow studies. No correlation was found between the grade of obstruction and anamnestic symptom data, the size of the prostate or residual urine. A slight correlation was found between the Schafer grade of obstruction and the flow curve pattern or peak flow. PMID- 8719366 TI - A long-term follow-up of 108 vasectomized men. Good counselling routines are important. AB - Contraceptive sterilization of men and women was made legal in Sweden in 1976. Males presently account for approximately 20% of the 8-10,000 people sterilized yearly in the country. Only few long-term follow-ups have been performed on Swedish men. Since 1983 preoperative counselling of men has been available in the county of Blekinge at family planning clinics. Prior to this counselling was available only at urologic or surgical departments. To evaluate long term satisfaction and the new counselling possibilities, all men vasectomized in Blekinge during 1985-1986 (n = 108) were investigated by a mailed questionnaire 2 years and 7 years after the operation. Response rates were 98% after 2 years and 96% after 7 years. 93% of the responding men were satisfied with their vasectomy when asked after 2 years. 95% reported satisfaction 5 years later. Problems mentioned concerned sexual performance, long lasting pain, and wanting children in new relationships. Regret was not encountered among the 25% who had been counselled at family planning centers. PMID- 8719367 TI - Multiple primary malignant neoplasms in urologic patients. AB - One hundred and four patients with multiple primary malignant neoplasm (MPN) with at least one genitourinary organ involved were analyzed. The 104 patients represented 0.9% of all cancer patients and 9.0% of all urologic tumor cases. Tumors of the prostate and bladder in urologic cancers and tumors of the gastrointestinal tract, particularly stomach cancer, in non-urologic cancers were predominantly involved. In 79 cases (76.0%), the second tumors were discovered within 5 years from the diagnosis of the first cancer. Renal cell carcinoma tended to be discovered incidentally and concomitantly. In contrast, bladder cancer tended to be an initial primary tumor presenting with symptoms, and tumor of prostate tended to be found incidentally, as a second cancer in metachronous cancer associations. All second tumors of the liver were found incidentally with ultrasonographic examination after a small interval. Strict follow-up at regular intervals at least the initial 5 years are essential for the detection of second malignant neoplasms. PMID- 8719368 TI - The prognostic value of p53 in superficially infiltrating transitional cell carcinoma. AB - Approximately 1/3 of T1 bladder cancers treated by endoscopic resection alone will progress. Prognostic factors are needed to help selecting appropriate treatment for these tumors. The purpose of the present investigation was to study the relation between p53 nuclear overexpression and disease progression. Tumors from 59 patients were studied by means of immunohistochemical nuclear staining. Forty tumors showed p53 nuclear overexpression while 19 tumors scored negative for mutations. We could not demonstrate any correlation between mutations of the p53 protein and tumor grading. p53 nuclear overexpression was not related to disease progression. PMID- 8719369 TI - Quality of life, sexual functioning and sex role identity after surgical orchidectomy in patients with prostatic cancer. AB - Surgical orchidectomy is a simple procedure with few physical side effects, low mortality, and cost effectiveness. Nevertheless, there can be negative sequelae such as sexual dysfunction, impaired quality of life, and poor body image. Although it is a frequent treatment approach for prostate cancer, it is not clear whether these sequelae are problematic for this patient group. It is possible that relief from painful metastases and the prolongation of life outweigh these negative factors. The present study investigated quality of life, sex-role identity, and sexual function in 15 patients with stage D prostate cancer, before and after surgery. Orchidectomy did not appear to affect quality of life, or sex role identity. However, loss of sexual function did present as an area of concern. It was noted that 55% of premorbidly sexually active patients found this loss disturbing. These patients, premorbidly, appeared to have higher sex-role stereotypy. PMID- 8719370 TI - Accuracy of clinical staging in non-seminomatous testicular cancer--a single centre experience of retroperitoneal lymph node dissection. AB - The management of low stage non-seminomatous testicular cancer remains a controversial issue. Programs of surveillance or primary retroperitoneal lymph node dissection (RPLND) after orchiectomy show equally good survival rates. Current focus is therefore on reduction of toxicity or side effects of the treatment while maintaining maximal prognostic safety. The clinician's decision of therapy is based on clinical staging methods including computerized tomography, pulmonary x-rays and serum tumour marker levels. In this study, the accuracy of clinical staging was compared with histopathology in 64 patients with clinical stages (CS) I and IIa, operated upon with RPLND between 1980 and 1992. Lymph node metastases were histopathologically verified in 37% of CS I and in 47% of CS IIa tumours. Thus, the clinical staging was inaccurate in 37% in CS I and in 53% in CS IIa patients. No clear relationship was shown between the risk factors: vascular invasion and/or tumour marker levels and metastatic spread. The specificity of clinical staging in non seminomatous testicular cancer was low. RPLND, on the other hand, is a reliable method for assessment of metastatic spread and will minimise unnecessary use of chemotherapy. Modern techniques for lymphadenectomy have a very low rate of post-operative morbidity. Development of better non-invasive imaging techniques for detection of lymph node metastases is hoped for, in order to improve the information on tumour spread and make it possible to individualize therapy. Thus, unnecessary therapy and following side effects can be avoided, improving the patient's quality of life during and after treatment. PMID- 8719371 TI - Recurrent hemolytic uremic syndrome in a renal transplant patient during pregnancy. AB - The recurrence of the hemolytic uremic syndrome (HUS) in a renal transplant patient during pregnancy and during treatment with cyclosporin is reported. After 4 1/2 months of treatment with hemodialysis the renal function partially recovered. A monozygote twin sister has not developed HUS in spite of 2 pregnancies. PMID- 8719372 TI - Pyeloureteral junction obstruction in upper moiety of duplex system. AB - A case of pyeloureteral junction obstruction in the upper moiety of a duplex system is reported. Pathologic, clinical and diagnostic aspects of this rare condition are discussed. PMID- 8719373 TI - Maximal displacement of the ureter by a liposarcoma. AB - We describe a patient with displacement of both ureters to the right side caused by a liposarcoma. The left ureter was displaced beyond the right ureter. The risk of intraoperative trauma to the ureter could be minimized by preoperative placement of ureteral stents. PMID- 8719374 TI - Fluid absorption during transurethral bladder surgery. AB - We report 2 patients who absorbed large amounts of irrigating fluid during transurethral resection of bladder tumours (TURB). Ethanol indication facilitated diagnosis of this complication and showed that absorption had occurred by the extravascular route. Symptoms occurred in one case but they resolved when hypertonic saline and furosemide were given. PMID- 8719375 TI - Seminal vesicle abscess: a case report and review of literature. AB - Seminal vesicle abscess, a rare urologic entity, is usually diagnosed with computerized tomography (CT) scan. We report a case of seminal vesicle abscess that required a transrectal ultrasonography to confirm the diagnosis. An analysis of 20 cases of seminal vesicle abscess reported in the literature is also reviewed. PMID- 8719376 TI - Prostatic abscess in a child. AB - We report a metastatic prostatic abscess in a 12-year-old boy. Rectal examination revealed a tender, fluctuating prostatic mass, causing acute retention of urine. Prostatic imaging by computerized tomography and transrectal ultrasonography were important in the diagnosis and management. PMID- 8719377 TI - Plaque sealing by coronary angioplasty. PMID- 8719378 TI - Effects of technique modification on immediate results of high speed rotational atherectomy in 710 procedures on 656 patients. AB - Seven hundred ten high speed rotational atherectomy (HSRA) procedures were performed in a single consecutive series of 656 patients. Stand alone HSRA was performed in 253 patients (35%). HSRA with adjunctive low pressure (< or = 2 ATM) balloon angioplasty (LP BA) was performed in 221 patients (31%), and HSRA with adjunctive high pressure (> or = 4 ATM) balloon angioplasty (HP BA) was performed in 236 patients (34%). Prognostically unfavorable Type B2 and C lesions dominated the study group (74.7%). Procedural success rate was 96%. Emergency coronary artery bypass surgery was performed in 1.4% of cases, Q wave myocardial infarction occurred in 3.4% and death, related to procedure, was consequent in 0.5% of cases. Incidence of flow limiting dissections was 3.1%, distal spasm was 5.3%, and "no reflow" phenomenon was 1.8%. The recent technique modifications included continuous advancer/guiding catheter infusion of the nitroglycerin verapamil mixture, limitation of duration of lesion engagement by the burr, stepwise increase in the burr size, decrease of rotational speed, and strict control of rpm drop during lesion ablation. Evolution of the interventional technique involved trends towards decrease of the use of HP BA in conjunction with steady increase in the percentage of SA and LP BA procedures over time. These technique changes resulted in complete absence of "no reflow" in 1994, as well as a generalized decrease in overall coronary vascular reactivity from all burr passes. PMID- 8719379 TI - Rotablator technique and complications? PMID- 8719380 TI - Coronary air embolism: incidence, severity, and suggested approaches to treatment. AB - Because no well-controlled study of inadvertent coronary air embolism has been done to truly quantify the incidence of this cardiac catheterization complication, we wanted to determine its incidence and severity in an active teaching medical center and assess approaches to treatment. We retrospectively reviewed 3,715 coronary angiogram and PTCA reports performed over 32 months. Further, we classified severity based on angiographic findings and symptoms as minimal, mild, moderate, and massive. Two independent angiographers reviewed 764 consecutive cines performed in the first 2 months of training of each new fellow and 740 cines performed in the last 2 months of training. We found that during the first 2 months of training the overall incidence for significant intracoronary air embolism was 0.19% (7 documented cases) compared with 0.2% (3 cases) for non-reported, minimal asymptomatic air embolism. The estimated incidence for total air emboli events was 0.27% (10/3,715). We did not find coronary air emboli in the 740 cines performed at the end of fellowship training. Additionally, the incidence of coronary air emboli during PTCA training was much higher compared with coronary angiography training (0.84 vs. 0.24%). Although there is no best technique to restore blood flow after blockage by air emboli, we suggest as options aspirating the air or forcefully injecting saline, with auxiliary supportive measures like 100% oxygen, IABP, CPR, and DC cardioversion. PMID- 8719381 TI - To air is human, to prevent is divine. PMID- 8719382 TI - Abnormal diastolic unopacified blood front: a useful marker of mitral stenosis. AB - The left ventriculograms of 113 patients, divided into seven groups, were reviewed for the detection of unopacified front (UOF) of blood entering the left ventricle (LV) during diastole. Normal UOF was detected in all the patients without left-sided valvular heart disease (regardless of the existence of coronary artery disease), constrictive pericarditis, aortic stenosis, and mitral regurgitation, although in the last group the UOF faded after several beats in 70% of cases. Only 5% of the patients with isolated mitral stenosis (MS) had an UOF, 75% had an abnormal UOF (< 60% of LV inflow tract), and 20% (with the greatest degree of MS) had only a dilution effect. A dilution effect was found in 62.5% of the patients with severe (> or = 3 degrees) aortic regurgitation (AR). An abnormal UOF is a sensitive and specific marker of significant MS. A dilution effect is indicative of either a significant MS or severe AR. PMID- 8719383 TI - Recanalization of totally occluded saphenous vein grafts using local urokinase delivery with the Dispatch catheter. AB - Current techniques for the percutaneous revascularization of totally occluded vein grafts are limited by a low initial success rate, a significant incidence of distal embolization, and a high rate of early graft reclosure. This case report describes two patients in whom graft recanalization was attempted with the combined use of balloon angioplasty/intra-graft stent placement and local urokinase delivery using a new angiotherapy catheter. Successful recanalization was achieved in both patients without major complications, in spite of a large thrombus burden as demonstrated by angiography. PMID- 8719384 TI - Recanalization of occluded SVGs: is there light at the end of the graft? PMID- 8719386 TI - Precision angioplasty. PMID- 8719387 TI - Extraction atherectomy for the recanalization of totally occluded aortocoronary saphenous vein grafts. AB - This report describes the successful treatment of totally occluded aortocoronary saphenous vein grafts with percutaneous transluminal extraction atherectomy (TEC) in three patients with severe angina and high reoperative surgical risk. The method presented here may provide an alternative to overnight urokinase infusion or repeat surgery in high risk patients. PMID- 8719385 TI - Efficacy of angioscopy in determining the effectiveness of intracoronary urokinase and TEC atherectomy thrombus removal from an occluded saphenous vein graft prior to stent implantation. AB - Percutaneous revascularization of thrombus containing saphenous vein grafts is associated with a high incidence of acute complications. This case report describes successful revascularization of an occluded vein graft employing angioscopically guided sequential urokinase infusion, TEC atherectomy and stent implantation. PMID- 8719388 TI - Translesional pressure and flow velocity after thrombolysis: assessment of suboptimal angioplasty for serial lesions. AB - Translesional pressure and flow velocity observations in the right coronary artery remain among the most difficult data to interpret because of variation in the location and relative size of branches, and unsuspected diffuse disease. Use of distal hyperemic response and translesional pressure gradients provides the most accurate assessment of serial lesions in such patients. In addition, the use of intracoronary thrombolysis can improve the angiographic appearance of lesions and facilitate later successful interventions. The post-infarction distal microvascular responsiveness may be impaired and, thus, abnormal coronary reserve values in this particular setting should be considered with lesion specific indicators of successful recanalization. PMID- 8719389 TI - New technique for exchanging an "over-the-wire" balloon dilatation catheter. AB - We describe here an indigenous technique of "over-the-wire" balloon exchanges that avoids use of an extension guidewire. PMID- 8719390 TI - Diffuse vasospasm following stenting of a free gastroepiploic artery graft: resolution with balloon angioplasty and intensive medical therapy. AB - Vasospasm following balloon angioplasty of gastroepiploic artery bypass grafts can be prevented or reversed with vasodilators. In our patient, stent deployment for ostial stenosis of a free gastroepiploic artery graft was accompanied by severe, diffuse spasm and a change in graft configuration that required both intensive medical therapy and balloon angioplasty for resolution. PMID- 8719391 TI - Coronary angioplasty of branch vessels associated with an extreme angle take-off. AB - Stenoses in branch vessels which originate at acute angle to the main vessel represent a technically challenging anatomy for the performance of balloon angioplasty [PTCA]. We report a novel technique utilizing a distally placed perfusion balloon that facilitated guidewire placement and subsequent balloon angioplasty of a branch vessel stenosis. PMID- 8719392 TI - Successful treatment of chronic total peripheral occlusions that failed conventional techniques using the stiff backend of the Glidewire. AB - The vast majority of failures of transcatheter interventions in patients with peripheral vascular disease are due to inability to cross the lesion with a guidewire. Although the use of the Glidewire has clearly improved the success rate, failures with especially chronic total occlusions still occur. We describe a new technique using the very stiff backend of the Glidewire, which we have found very successful, when conventional techniques fall in crossing highly resistant lesions. PMID- 8719393 TI - Strategies for the palliation of severe unprotected left main coronary artery disease: use of newer technologies. AB - Angioplasty of unprotected left main coronary artery lesions is associated with high procedural and late mortalities. We describe the successful use of rotational coronary atherectomy with prophylactic supportive measures in the management of a heavily calcified unprotected left main lesion. This report demonstrates that high-risk lesions, previously regarded as unapproachable, may be safety treated with newer technologies. PMID- 8719394 TI - Reconsidering unprotected left main coronary angioplasty. PMID- 8719395 TI - Evidence for dynamic coronary vasoconstriction of non-infarct vessels during acute myocardial infarction. AB - The role of coronary vasoconstriction in acute myocardial infarction is controversial. Two patients are presented who were studied with angiography during the early hours of acute myocardial infarction and in whom reversible spasm of non-infarct vessels was observed. A tendency towards coronary vasospasm may be a generalized phenomenon during acute myocardial infarction. PMID- 8719396 TI - "Results of bailout coronary stenting". PMID- 8719397 TI - Delayed appearance of traumatic aorto-right ventricular fistula. PMID- 8719398 TI - Balloon mitral valvuloplasty: selection of the technique. PMID- 8719399 TI - "Functional characteristics of a new perfusion balloon: comparison with the Flowtrack 40 in a closed chest swine". PMID- 8719400 TI - Simultaneous inflation of tandem (sequential) angioplasty balloons. PMID- 8719401 TI - Ectopic origin of the right coronary artery: anterior or posterior--that is the question. PMID- 8719402 TI - Is there more radiation using a two-monitor system? PMID- 8719403 TI - Atrial septal puncture for balloon mitral valvotomy. PMID- 8719404 TI - Inadvertent pulmonary arterial hypertension induced by Berman angiography catheter. PMID- 8719405 TI - [3H]1-aminocyclopropanecarboxylic acid, a novel probe for strychnine- insensitive glycine receptors. AB - [3H]1-Aminocyclopropanecarboxylic acid (ACPC) exhibits high affinity, specific binding to strychnine-insensitive glycine receptors. In extensively washed rat forebrain membranes, the specific binding of [3H]ACPC was optimal at 25 degrees C in the presence of 10 mM MgCl2. Comparable levels of specific [3H]ACPC binding were obtained using centrifugation and filtration for separation of bound from free radioligand. [3H]ACPC labels two sites with Kdl and Bmax1 values of 129 +/- 34 nM and 2.30 +/- 0.37 pmol/mg protein and Kd2 and Bmax2 values of 7.26 +/- 1.69 microM and 20.6 +/- 2.2 pmol/mg protein for the high and low affinity sites, respectively. The Kd of [3H]ACPC (66 nM) estimated under non-equilibrium conditions (koff = 8.91 +/- 0.78 x 10(-3) s-1; kon = 1.35 x 10(-4) nM-1 s-1) was similar to the value obtained for the high affinity site obtained by equilibrium binding. The Kd1 of[3H]ACPC is in good agreement with the previously reported Ki values of ACPC to inhibit the binding of other glycinergic ligands including [3H]glycine, [3H]5,7-dichlorokynurenic acid (5,7-DCKA) and [3H]L-689,560 ((+/-)-4 (trans)-2-carboxy-5,7-dichloro-4- phenylaminocarbonylamino-1,2,3,4 tetrahydroquinoline). Moreover, the potencies of a series of glycine site ligands, including glycine. ACPC, 1-aminocyclobutanecarboxylic acid (ACBC), 5,7 DCKA, 7-chlorokynurenic acid (7-CKA), R(+)-3-amino-1-hydroxy-2- pyrrolidine (HA 966) and D-serine, to inhibit [3H]ACPC binding were highly correlated with their potencies to inhibit [3H]glycine and [3H]5,7-DCKA binding (r2 = 0.98-0.51). These results demonstrate that [3H]ACPC is a useful tool for examining the neurochemical and pharmacological properties of strychnine-insensitive glycine receptors. PMID- 8719406 TI - Differential effects of lectins on recombinant glutamate receptors. AB - The effects of the lectins concanavalin A, succinyl concanavalin A, wheat-germ agglutinin and soybean agglutinin were studied at recombinant ionotropic glutamate receptors expressed in Xenopus oocytes. Homomeric and heteromeric receptors from each of the three major classes of ionotropic glutamate receptors (N-methyl-D-asparate (NMDA), alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) and kainate) were studied. The lectins potentiated homomeric configurations of kainate, AMPA and NMDA receptors to a greater degree than the corresponding heteromeric configurations although the rank order of the lectin potentiating effects was the same for both homomeric and heteromeric receptors within a given glutamate receptor class. The most profound effects of the lectins were observed with the kainate receptors; the rank order of potentiating effects of the lectins at the homo- and heteromeric kainate receptors (Glu6 and Glu6/KA 2) was concanavalin A > succinyl concanavalin A > wheat-germ agglutinin > soybean agglutinin. At the recombinant Glu3 and Glu2/3 AMPA receptor complexes, wheat germ agglutinin and concanavalin A produced the largest enhancements of the glutamare-activated currents followed by succinyl concanavalin A; soybean agglutinin had no significant potentiating effect. Agonistevoked currents recorded from oocytes expressing the homo- and heteromeric NMDA receptors were only slightly enhanced by concanavalin A and succinyl concanavalin A but not by wheat-germ agglutinin or soybean agglutinin. These results demonstrate that kainate. AMPA and NMDA receptors display dramatic differences in their responses to lectins, and suggest that the receptor-bound oligosaccharide side chains may play different roles in the functional responses mediated by the three major classes of ionotropic glutamate receptors. PMID- 8719407 TI - Effects of benfluorex metabolites on membrane fluidity and insulin-related processes. AB - As little work has dealt with the antihyperglycemic property of benfluorex at the hepatocyte level, we studied the effects of its main metabolites, S422 and S1475, on membrane fluidity and on insulin binding, internalization and action in healthy rat hepatocytes. Both metabolites were effective fluidizing agents. Neither one affected insulin binding. Only S422 favored the bound insulin receptor internalization process. The metabolites produced no change in basal alpha-aminoisobutyric acid uptake. Only S422 promoted the insulin-stimulated alpha-aminoisobutyric acid uptake in a dose-dependent way. Therefore, our study demonstrated that: (i) the effects of S422 on insulin-related processes in isolated hepatocytes were direct, specific and not due to any membrane fluidizing mechanism; (ii) S422 improved hepatocyte response to insulin at a post-binding level. These results in vitro give an additional explanation, at the cellular level, of the benefit of benfluorex treatment for non insulin-dependent diabetes patients. PMID- 8719408 TI - Additivity and non-additivity between dopamine-, norepinephrine-, carbachol- and GABA-stimulated GTPase activity. AB - The mode of coupling between neurotransmitter receptors and G proteins was investigated by agonist-induced high-affinity GTPase activity in rat striatal membranes. There was a simple additive relationship among dopamine-, carbachol-, and gamma-aminobutyric acid (GABA)-sensitive high-affinity GTPase activity in any combination, indicating that the respective receptors stimulated by these agonists (i.e., dopamine D2, pirenzepine-insensitive muscarinic, and GABAB receptors) interact independently with distinct pools of G proteins. Unexpectedly non-additivity was observed between dopamine- and norepinephrine-stimulation. This lack of additivity was apparently due to stimulation of the same dopamine D2 receptors by both dopamine and norepinephrine, since norepinephrine-stimulated high-affinity GTPase activity could be inhibited by dopaminergic but not adrenergic antagonists. The same non-additivity as seen in rat striatum was confirmed in the membranes prepared from cultured mouse fibroblast cells co transfected with dopamine D2 and adenosine A2A receptors. The implication of the (non-)additivity between receptor-mediated high-affinity GTPase activity was discussed with a consideration of the possible underlying molecular mechanism. PMID- 8719409 TI - Structural dependence of the allosteric interaction of semi-rigid verapamil analogues with dihydropyridine-binding in kitten heart. AB - Structural determinants of the allosteric interaction of semi-rigid verapamil analogues with dihydropyridine binding were investigated in kitten heart using [3H](+)-isradipine as radioligand. Chemical variations were performed in the alkyl chain of verapamil and include introduction of unsaturation (double or triple bonds) or the insertion of cyclohexyl moieties. Introduction of unsaturation generally reduces the allosteric interaction in the case of 'double bond'-and abolishes it in the case of 'triple bond'-derivatives. Also the introduction of cyclohexyl moieties diminishes the potency of allosteric interaction: derivatives with the phenylethylamino side chain in an equatorial position exhibit the allosteric interaction, while it is lacking in derivatives with the basic side chain in axial position. Thus, the reduced conformational flexibility of the new verapamil congeners reduces or abolishes their ability to allosterically interfere with dihydropyridine binding. A molecular interpretation was approached by molecular modelling studies. The strategy was to find low energy conformations common to the active congeners, but not shared by the inactive ones. Structural features discriminating allosterically active and inactive congeners comprise: 1) the position of the nitrogen, 2) the volume occupied by the N-methyl groups, 3) the direction of the N-H bond and 4) the position of the phenyl ring in the basic side chain. PMID- 8719410 TI - Effects of caffeine on Ca2+ fluxes and secretion in bovine chromaffin cells. AB - The effects of caffeine on Ca2+ fluxes and catecholamine secretion in bovine adrenal chromaffin cells were examined. Caffeine inhibited secretion. 45Ca2+ uptake and cytosolic Ca2+ concentration ([Ca2+]i) rise induced by the nicotinic receptor agonist 1.1-dimethyl- 4-phenylpiperazinium (DMPP) and the Na+ channel activator veratridine. The inhibitory effect of caffeine on high K(+)-induced secretion was smaller than that on DMPP- and veratridine-induced responses. Caffeine only slightly inhibited high K(+)-induced 45Ca2+ uptake and did not affect [Ca2+]i rise. Caffeine also inhibited muscarinic receptor-mediated inositol phosphate generation. Our results suggest that the inhibitory effects of caffeine on bovine chromaffin cells mainly occur at both muscarinic and nicotinic receptors as well as at the voltage-dependent Na+ channels and to a smaller extent at site(s) distal to Ca2+ entry. The effects of caffeine on nicotinic receptors but not on muscarinic receptors can be explained by its ability to raise intracellular cAMP. PMID- 8719411 TI - Pre-incubation of synaptosomes with arachidonic acid potentiates inhibition of [3H]D-aspartate transport. AB - The ability of low micromolar concentrations of the polyunsaturated fatty acid, arachidonic acid (cis-5,8,11,14-eicosatetraenoic acid) to inhibit the high affinity, sodium-dependent transport of [3H]D-aspartate into purified synaptosomes of rat brain has been examined. Pre-incubation of the synaptosomes with arachidonic acid for 10-60 min produced a marked potentiation of the response to 10 microM arachidonic acid compared to co-incubation, and the threshold for inhibition of [3H]D-aspartate transport occurred at a concentration of 1 microM. Minimal inhibition of transport was seen with the unsaturated fatty acids, cis-oleic (cis-9-octadecenoic acid) and cis-linolenic (cis-9,12,15 octadecatrienoic acid), nor with the 20-carbon saturated fatty acid, arachidic acid (n-eicosanoic acid). Inclusion of the cyclo-oxygenase inhibitor, nor dihydroguaretic acid (NDGA), in the presence of 5 microM arachidonic acid did not alter the inhibition of [3H]D-aspartate transport between 0-10 min, but did enhance the response at longer pre-incubation times. Inhibition of [3H]D aspartate transport by arachidonic acid persisted during addition of the calcium ionophore, A23187, whereas removal of calcium ions from the incubation medium potentiated the response to arachidonic acid. The results are discussed in terms of the physiological relevance of the inhibition of glutamate transport by arachidonic acid, and suggest that regulation of inhibition of the glutamate transporter by arachidonic acid may be achieved by changes in the extracellular, as well as the intracellular, concentration of calcium ions. PMID- 8719412 TI - Characterisation of a recombinant P2Y purinoceptor. AB - We have previously cloned a cDNA encoding a G-protein-coupled P2 purinoceptor from chick brain and designated this as a P2Y1 purinoceptor (Webb, T.E., J. Simon, B.J. Krishek, A.N. Bateson, T.G. Smart, B.J. King, G. Bumstock and E.A. Barnard, 1993, FEBS Lett. 324, 219). Here, we describe the further characterisation of this recombinant receptor expressed in both simian kidney endothelial (COS-7) cells and Xenopus oocytes. In transfected COS-7 cell membranes, the recombinant receptor showed a high level of expression (Bmax = 7.9 +/- 2.2. pmol [35S]dATP alpha S bound/mg protein) and affinity (Kd = 6.6 +/- 0.3 nM). In these COS-7 cells, the activation of the implanted purinoceptor induced a suramin-sensitive formation of inositol 1,4,5-triphosphatic (1,4,5InsP3). Upon expression in Xenopus oocytes, ATP was the only natural nucleoside triphosphate to elicit a Ca(2+)-activated chloride current. The P2 purinoceptor antagonists suramin and Reactive Blue-2 were both able to inhibit this evoked current. Utilizing both expression systems, the binding affinity profile and the functional pharmacological profile of the agonists, the common series found was: 2-methylthioATP (2-MeSATP) > or = ATP > ADP beta S > ADP. These two agonist series and the lack of activity of adenosine, alpha, beta-methyleneATP (alpha, beta-meATP), 3'-O-(4-benzoyl) benzoyl-ATP (Bz-ATP) and UTP, together confirmed that this receptor is a specific subtype of the P2Y purinoceptors. PMID- 8719413 TI - Influence of alpha-helicity, amphipathicity and D-amino acid incorporation on the peptide-induced mast cell activation. AB - Mast cell activation by polycationic substances is believed to result from a direct activation of G protein alpha subunits and it was suggested that the adaption of amphipathic, alpha-helical conformations would allow the peptide to reach the cytosolic compartment to interact with G proteins (Mousli et al., 194, Immunopharmacology 27, 1, for review). We investigated the histamine-releasing activity of model peptides as well as analogues of magainin 2 amide and neuropeptide Y with different amphipathicities and alpha-helix content on rat peritoneal mast cells. Amphipathic helicity is not a prerequisite for mast cell activation. Moreover, non-helical magainin peptides with high histamine-releasing activity were less active in the liberation of carboxyfluoresceine from negatively charged liposomes, indicating that peptide-induced mast cell activation and peptide-induced membrane perturbation do not correlate. In contrast to the negligible influence of the secondary structure, amino acid configuration may exert a striking influence on peptide-induced mast cell activation. Thus histamine-release by substance P was markedly impaired when the L-amino acids in the positively charged N-terminal region were replaced by D amino acids, with [D-Arg1)substance P being the most inactive substance P diastreoisomer. PMID- 8719414 TI - Expression and pharmacology of human GABAA receptors containing gamma 3 subunits. AB - A cDNA encoding the gamma 3 subunit of the human GABAA receptor has been obtained by molecular cloning. Its deduced amino acid sequence shows a high level of sequence identity with the published mouse and rat sequences (96%). The ligand binding pharmacology of the benzodiazepine site formed by stably-expressed human alpha 5 beta 3 gamma 2S and alpha 5 beta 3 gamma 3 GABAA receptor subtypes have been compared for a number of ligands, Benzodiazepine site ligands were found to be either non-selective or gamma 2-selective, with the exception of CL218,872, which was found to be 10-fold selective for the alpha 5 beta 3 gamma 3-containing subtype Two benzodiazepine site ligands. Ro15-4513 and FG8205 were more efficacious at alpha 5 beta 3 gamma 3 receptors than alpha 5 beta 3 gamma 2 receptors expressed in Xenopus oocytes, CL218,872, which is a partial agonist at alpha 1 containing receptors, had no intrinsic activity at either alpha 5 beta 3 gamma 2 or alpha 5 beta 3 gamma 3, alpha 1 beta 2 gamma 2S and alpha 1 beta 2 gamma 3 human GABAA receptors were also expressed in Xenopus oocytes and their benzodiazepine pharmacology investigated. Both the EC50 and efficacy of benzodiazepine site ligands were influenced by the type of gamma subunit coexpressed with alpha 1 and beta 2. PMID- 8719415 TI - Amidines are potent inhibitors of nitric oxide synthases: preferential inhibition of the inducible isoform. AB - We evaluated the ability of simple alkyl amidines to inhibit the activity of the inducible isoform of nitric oxide (NO) synthase in vitro. In immunostimulated J774 macrophages, 2-iminopiperidine (EC50 = 10 microM) and butyramidine (EC50 = 60 microM) were more potent than NG-methyl-L-arginine (EC50 = 70 microM) in inhibiting nitrite formation. The five amidines tested for their ability to inhibit the conversion of L-arginine to L-citrulline by bovine endothelial cell homogenates (a source of the constitutive, endothelial NO synthase isoform) were less effective than NG-nitro-L-arginine or NG-methyl-L-arginine. The rank-order of the potencies of the amidines against the endothelial NO synthase was, in general, similar to the rank-order of the pressor effects of these agents in anesthetized rats. Thus, certain amidines are potent inhibitors of NO synthase, and are more selective towards the inducible NO synthase than the commonly used L arginine based NO synthase inhibitors. PMID- 8719416 TI - Cloning and characterization of the human GABAA receptor alpha 4 subunit: identification of a unique diazepam-insensitive binding site. AB - Benzodiazepines modulate gamma-aminobutyric acid (GABA)-evoked chloride currents through a specific binding site at the GABAA receptor-chloride channel complex. The heterogeneity of diazepam-sensitive benzodiazepine binding sites (type I and type II) has been identified by pharmacological approaches both with native receptors and recombinant receptors coexpressing alpha, beta and gamma subunits. In addition, two distinguishable diazepam-insensitive benzodiazepine sites are found, spatially distributed between cerebral cortical and cerebellar regions. Coexpression of alpha 6 with beta 2 and gamma 2L subunits creates a pharmacologically similar benzodiazepine receptor to the diazepam-insensitive site observed in cerebellum, however, there is no evidence regarding the possible subunit combination forming the DI site in cerebral tissues. Here we report the cloning of the human alpha 4 cDNA and its pharmacology by coexpression of this alpha 4 subunit with beta 2 and gamma 2L subunits. This recombinant receptor complex showed a high affinity for the previously described benzodiazepine partial agonist bretazenill, the pyrazoloquinoline compounds CGS-9895 and CGS 9896, as well as the inverse agonists DMCM (methyl 6,7-dimethoxy 4-ethyl-beta carboline-3-carboxylate) and Ro15-4513 as determined by [3H]Ro15-4513 binding. However, it is insensitive to the benzodiazepine type I selective compounds CL218.872 (3-methyl-6-[3-(trifluoromethyl)[phenyl]-1,2,4-triazolo[4.3 b]pyridazine ) and zolpidem as well as the benzodiazepine full agonists diazepam, halazolam and midazolam. In addition, the benzodiazepine receptor ligands DMCM, beta-CCE (beta-carboline-3-carboxylate ethyl ester), Beta-CCM (beta-carboline-3 carboxylate methyl ester), FG-7142, CGS-9895 and CGS-9896 showed 7 to 10 times higher affinity for alpha 4 beta 2 gamma 2L. The pharmacology of the alpha 4 beta 2 gamma 2L receptor complex appears to resemble those of the diazepam-insensitive site found in the cerebral cortex. Our study thus suggests that this subpopulation of diazepam-insensitive GABAA receptors may be composed of alpha 4 beta 2 gamma 2L subunits. PMID- 8719417 TI - NS-49, a novel alpha 1a-adrenoceptor-selective agonist characterization using recombinant human alpha 1-adrenoceptors. AB - alpha 1-Adrenoreceptors comprise a heterogeneous family and subtype-selective ligands are valuable in studying the functional role of each receptor subtype. Using the Chinese hamster ovary (CHO) cells stably expressing the cloned human alpha 1-adrenoceptor subtypes (alpha 1a, alpha 1b, and alpha 1d)1, we have compared a newly synthesized phenethylamine class agonist (R)-(-)-3'-(2-amino-1 hydroxyethyl)-4'-fluoromethanesulfonanilide hydrochloride (NS-49) with imidazoline class agonist oxymetazoline in their binding affinities and intrinsic activities in causing transient increases of cytosolic Ca2+ concentrations ([Ca2+?i response). Radioligand binding studies with 2-[beta-(4-hydroxyl-3 [125I]iodophenyl)ethylamino-methyl]tetralone ([125I]HEAT) showed NS-49 and oxymetazoline had higher affinities at alpha 1a-than at alpha 1b- and alpha 1d subtypes (-log Ki values at alpha 1a-, alpha 1b-and alpha 1d-subtype: 6.18, 5.13, and 5.38 for NS-49: 8.19, 6.50, and 6.44 for oxymetazoline, respectively). In functional studies, both oxymetazoline and NS-49 worked as a selective and partial agonist at alpha 1a-subtype: however, NS-49 is more efficacious than oxymetazoline. NS-49 is the phenethylamine class of alpha 1-adrenoceptor partial agonist relatively selective and efficacious for the human alpha 1a-adrenoceptor subtype, NS-49 would be potentially useful for studying the physiological role of alpha 1-adrenoceptor subtype. PMID- 8719418 TI - Pituitary adenylate cyclase-activating polypeptide stimulates cardiodilatin/atrial natriuretic peptide (CDD/ANP-(99-126) secretion from cultured neonatal rat myocardiocytes. AB - It has been reported that the highly homologous neuropeptides pituitary adenylate cyclase-activating peptide (PACAP) and vasoactive intestinal polypeptide (VIP) exert similar cardiovascular effects in vivo. In the present study we compared the effects of these neuropeptides on myocardial cyclic AMP content and the release of immunoreactive CDD/ANP-(99-126) (atrial natriuretic peptide). In cultured neonatal rat cardiomyocytes PACAP and VIP evoke concentration-dependent increases in intracellular cyclic AMP content but responses to VIP are markedly less. PACAP stimulates the release of CDD/ ANP-(99-126) in a concentration dependent manner with a threshold concentration of 1 nM, and up to a 6-fold increase in basal secretion at 1 microM PACAP. In contrast. VIP had no effect on the release of CDD/ANP. Pretreatment of cells with the competitive PACAP antagonist, PACAP-6-38 (1 microM), significantly reduces the effects of PACAP on intracellular cyclic AMP and on CDD/ANP-(99-126) secretion and abolishes the effects of VIP on cyclic AMP. Pretreatment with VIP-receptor antagonist (1 microM) prevents the cyclic AMP-response to VIP while increases in cyclic AMP as well as stimulation of CDD/ANP-(99-126) release by PACAP are not affected. It is concluded that both neuropeptides directly influence cardiac myocytes through an increase in intracellular cyclic AMP. Release of CDD/ ANP-(99-126) by PACAP may be involved in the decrease in blood pressure that follows intravenous administration of this peptide. The higher potency of PACAP to induce cyclic AMP synthesis, its stimulating effect on the release of CDD/ANP-(99-126) and the finding that the VIP-receptor antagonist inhibits responses to VIP but not to PACAP suggest that PACAP activates cardiac myocytes through a PACAP-specific receptor. PMID- 8719419 TI - Effect of berbamine on cytosolic phospholipase A2 activation in rabbit platelets. AB - The effect of berbamine, a biscoclaurine alkaloid, on cytosolic phospholipase A2 activation in rabbit platelets was investigated. Berbamine inhibited arachidonic acid liberation induced by thrombin but not that by ionomycin. The alkaloid did not affect thrombin-stimulated Ca2+ mobilization. Ca(2+)-dependent translocation of cytosolic phospholipase A2 to membranes, or the activity of partially purified cytosolic phospholipase A2. Furthermore, berbamine had no effect on the thrombin elicited increase in cytosolic phospholipase A2 activity. However, berbamine suppressed arachidonic acid liberation in platelets stimulated with GTP-binding protein activators. Although incubation of platelet membranes with a GTP analogue decreased the islet-activating protein-catalyzed ADP-ribosylation of an approximately 40 kDa protein in the membranes, pretreatment of the membranes with berbamine did not influence the decrease in ADP-ribosylation. These results suggest that berbamine may impair GTP-binding protein-mediated activation of cytosolic phospholipase A2, probably without influencing the enzyme translocation to membranes or the increase in the enzyme activity, and thus may cause the suppression of thrombin-induced arachidonic acid liberation. PMID- 8719420 TI - Impairment of adenylyl cyclase signal transduction in mecobalamin-deficient rats. AB - This study examined alterations in the beta-adrenoceptor-G5-adenylyl cyclase system in cerebral cortex membranes from vitamin B12-deficient rats fed a diet lacking vitamin B12 (mecobalamin) for 15 weeks. Basal, 5(7) guanylylimidodiphosphate (GppNHp)-, isoproterenol-, and forskolin-stimulated adenylyl cyclase activities were significantly reduced in mecobalamin-deficient rats compared with those in control rats. However, no significant differences were observed in the amount and function of G5- estimated by immunoblotting and guanine nucleotide photoaffinity labeling, respectively, or in the densities and the dissociation constants of beta-adrenoceptors, estimated by [125I] pindolol binding, between control and the deficient rats. These results indicate that vitamin B12 deficiency results in the impairment of the coupling among the beta adrenoceptor, G5- and the catalytic subunit of adenylyl cyclase, and in dysfunction of the catalytic subunit of the enzyme, suggesting that vitamin B12 participates in the regulation of neuronal adenylyl cyclase signal transduction. PMID- 8719421 TI - Carazolol: a potent, selective beta 3-adrenoceptor agonist. AB - Carazolol is a beta1/beta2 adrenoceptor antagonist of high potency used in the treatment of hypertension. Its affinity for the beta 3-adrenoceptor was determined in Chinese hamster ovary cells transfected with the gene of the human or the murine beta 3-adrenoceptor. Carazolol is recognized with a nanomolar affinity, which positions it among the best ligands for beta 3-adrenoceptors. The adenylyl cyclase stimulation was measured in transfected cells where carazolol acted as a full agonist on both murine and human receptor subtypes. Furthermore, in murine adipocyte-like 3T3-F442A cells, which express beta 3-adrenoceptor naturally, carazolol induced lipolysis. This compound also appeared to be a useful tool for molecular characterization of the beta 3-adrenoceptor, unlike the classical beta 3-adrenoceptor agonists, carazolol conferred an appreciable protection of receptor binding sites against inactivation by the reducing agent dithiothreitol. The major iodinated analog of carazolol retained its binding characteristics for the beta 3-adrenoceptor and remained an efficient adenylyl cyclase stimulator in cells expressing human beta 3-adrenoceptor. PMID- 8719422 TI - Pharmacological study of dihydroetorphine in cloned mu-, delta- and kappa-opioid receptors. AB - We investigated the binding characteristics of dihydroetorphine, 7,8-dihydro-7 alpha-[1-(R)-hydroxy-1-methylbutyl]-6, 14-endoethanotetrahydro-oripavine, and its effect on the inhibitory system of cyclic AMP production using cloned mu-, delta- and kappa-opioid receptors expressed on Chinese hamster ovary cells. The Ki values of dihydroetorphine for the mu-, delta- and kappa-opioid receptors were 4.5 x 10(-10). 1.8 x 10(-9) and 5.7 x 10(-10) M, respectively. On the other hand, those of morphine were 1.9 x 10(-9), 1.4 x 10(-6) and 1.3 x 10(-7) M, respectively. Through all of these three types of opioid receptors, dihydroetorphine inhibited forskolin (10 microM)-stimulated cyclic AMP production via pertussis toxin-sensitive G protein(s), and the inhibitory effects were antagonized by co-application with opioid receptor antagonists. The IC50 values of dihydroetorphine for the inhibition of cyclic AMP production through the mu-, delta- and kappa-opioid receptors were 4.2 x 10(-11), 8.6 x 10(-10) and 4.3 x 10( 9) M. respectively. On the other hand, those of morphine were 2.6 x 10(-8), 2.6 x 10(-6) and 1.9 x 10(-6) M, respectively. These results indicate that dihydroetorphine, unlike morphine which preferentially binds the mu-opioid receptor, binds not only mu- but also delta- and kappa-opioid receptors with high affinity and acts as a more potent agonist than morphine for all of the three types of receptors. PMID- 8719423 TI - Nitric oxide modulation of Ca2+ responses in cow tracheal epithelium. AB - Nitric oxide (NO) plays an important role in the regulation of Ca2+ -dependent airway epithelial function such as ciliary motility. In this experiment, the effect of NO on intracellular Ca2+ ([Ca2+]1) was studied in cultured cow tracheal epithelium by the fura-2 method. L-NG-nitroarginine methyl ester, an NO synthase inhibitor, per se did not significantly alter baseline [Ca2-]i, but bradykinin- and ATP-induced increases in [Ca2+]i were significantly reduced in the presence of L-NG-nitroarginine methyl ester. This inhibitory effect disappeared by a simultaneous addition of L-arginine. Sodium nitroprusside or dibutyryl cyclic GMP potentiated bradykinin- and ATP-induced increases in [Ca2+]i. Cytochemistry for NADPH diaphorase activity revealed the presence of NO synthase in the cultured epithelium. These results suggest that NO produced by NO synthase in airway epithelium modulates bradykinin- and ATP-induced [Ca2+]i responses, which may be dependent on cyclic GMP. PMID- 8719424 TI - Pharmacological studies on mechanisms involved in Ca2+ oscillations in rat megakaryocytes. AB - Extracellular application of ATP evoked the oscillatory K+ currents (IKCa) reflecting oscillation in cytoplasmic Ca2+ concentration ([Ca2-]i) of megakaryocyte isolated from rat bone marrow. We have reported that the [Ca2+], oscillation was regulated by intracellular Ca(2+)-pumping activity (Uneyama H.C. Uneyama and N. Akaike, 1993, J. Biol. Chem. 268, 168). Here we found that the Ca2+ pump of the megakaryocyte could be divided into at least two classes according to the sensitivity to phosphorylation-modulating drugs. The effects of protein kinase C and cyclic AMP-dependent protein kinase are complementary, and the effect of Ca2+/calmodulin is independent of the above two kinases. In addition, this is the first report concerning the physiological regulation of Ca(2+)-ATPase in living cells. PMID- 8719426 TI - Potentiation and inhibition of subtypes of neuronal nicotinic acetylcholine receptors by Pb2+. AB - Effects of inorganic lead (Pb2+) on defined subtypes of neuronal nicotinic acetylcholine receptors have been investigated. Voltage clamp experiments have been performed on Xenopus oocytes expressing alpha 3 beta 2, alpha 3 beta 4 and alpha 4 beta 2 neuronal nicotinic acetylcholine receptor subunit combinations. In oocytes expressing the alpha 3 beta 2 subunit combination Pb2+ enhances the peak amplitude of nicotinic acetylcholine receptor-mediated inward currents evoked by superfusion with 100 microM acetylcholine. At concentrations of 1-250 microM, Pb2+ potentiates alpha 3 beta 2 receptor-mediated inward current concentration dependently by a factor of 1.1-11.0. Inward currents evoked by low (3 microM) and high (1 mM) concentrations of acetylcholine are potentiated to a similar extent. Conversely, in oocytes expressing the alpha 3 beta 4 subunit combination Pb2+ inhibits the nicotinic receptor-mediated inward currents evoked with 100 microM acetylcholine. Inhibitory effects are observed in the concentration range of 1 nM 100 microM Pb2+ but the degree of inhibition varies between oocytes. A similar inhibition of the alpha 4 beta 2 nicotinic receptor-mediated inward current by Pb2+ indicates that alpha as well as beta subunits are involved in the potentiating and inhibitory effects. Possible reasons for the variation in the inhibitory effects of Pb2+ on alpha 3 beta 4 and alpha 4 beta 2 nicotinic receptor-mediated inward currents have been investigated and are discussed. The divalent cations Ca2+ and Mg2+ potentiate both alpha 3 beta 2 and alpha 3 beta 4 nicotinic receptor-mediated inward currents. The distinct modulation of receptor function by Pb2+ and by Ca2+ and Mg2+ and the dependence of the modulatory effect of Pb2+ on subunit composition suggest that Pb2+ interacts with multiple sites on the alpha and beta subunits of neuronal nicotinic acetylcholine receptors. PMID- 8719425 TI - Ba2+ replaces Ca2+/calmodulin in the activation of protein phosphatases and in exocytosis of all major transmitters. AB - Exocytosis from nerve terminals is triggered by depolarization-evoked Ca2+ entry, which also activates calmodulin and stimulates protein phosphorylation. Ba2+ is believed to replace Ca2+ in triggering exocytosis without activation of calmodulin and can therefore be used to unravel aspects of presynaptic function. We have analysed the cellular actions of Ba2+ in relation to its effect on transmitter release from isolated nerve terminals. Barium evoked specific release of amino acid transmitters, catecholamines and neuropeptides (EC50 0.2-0.5 mM), similar to K-/Ca(2+)-evoked release both in extent and kinetics. Ba(2+)-and Ca(2+)-evoked release were not additive. In contrast to Ca2+, Ba2+ triggered release which was insensitive to trifluoperizine and hardly stimulated protein phosphorylation. These observations are in accordance with the ability of Ba2+ to replace Ca2+ in exocytosis without activating calmodulin. Nevertheless, calmodulin appears to be essential for regular (Ca(2+)-triggered) exocytosis, given its sensitivity to trifluoperizine. Both Ba(2+)-and Ca(2+)-evoked release were blocked by okadaic acid. Furthermore, anti-calcineurin antibodies decreased Ba(2+)-evoked release. In conclusion, Ba2+ replaces Ca2+/calmodulin in the release of the same transmitter pool. Calmodulin-dependent phosphorylation appears not to be essential for transmitter release. Instead, our data implicate both Ca(2+)-dependent and -independent dephosphorylation in the events prior to neurotransmitter exocytosis. PMID- 8719427 TI - Implication of dopamine transporter system on 1-methyl-4-phenylpyridinium and rotenone effect in striatal synaptosomes. AB - The neurotoxic effect of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) seems to be produced by the inhibition of the respiratory chain by its metabolite 1-methyl-4-phenylpyridinium ion (MPP+). At the same time, its specific selectivity seems to be related especially to the dopamine uptake system. However, it is possible that other specific differences in dopaminergic neurons at the nigrostriatal system, such as constitutive metabolic deficiencies or other differences related to the energy capacity, could determine the greater vulnerability to MPP+. We have addressed this point by studying the effect of MPP+ and different inhibitors of the respiratory chain (rotenone, antimycin A and KCN) on the maximal respiratory rate from both synaptosomes and isolated synaptosomal mitochondria from different brain areas, i.e. cortex, hippocampus and striatum, and in isolated liver mitochondria. The results demonstrate the absence of differences in the effect of the inhibitors in isolated mitochondria. In contrast, a greater inhibition was found in striatal synaptosomes than in cortical or hippocampal synaptosomes when MPP+ and rotenone were used. Moreover, nomifensine or 1-[2-[bis(4-fluorophenyl)methoxy]ethyl]-4-(3-phenylpropyl) piperazine dihydrochloride (GBR-12909), inhibitor of the dopamine uptake system, has a protective effect in both cases. Our study indicates the great importance of the dopamine uptake system in the vulnerability of the dopamine striatum system. Moreover, our results show the low selectivity of this dopamine uptake system that is able to transport actively compounds with different chemical structures such as dopamine, MPP+ and rotenone. PMID- 8719428 TI - A long-term receptor stimulation is requisite for angiotensin II-dependent DNA synthesis in vascular smooth muscle cells from spontaneously hypertensive rats. AB - Angiotensin II stimulates DNA synthesis in aortic smooth muscle cells prepared from spontaneously hypertensive rats, with maximal levels detected 20 h after stimulation. Angiotensin II receptor antagonists inhibited the angiotensin II induced DNA synthesis. In particular, the noncompetitive antagonist 2-ethoxy-1 [[2'(1 H-tetrazol-5-yl) biphenyl-4-yl]methyl]-1 H-benzimidazole-7-carboxylic acid (CV11974) was more effective than expected from its affinity for the angiotensin II receptor and its potency for inhibiting angiotensin II-induced increase in cytosolic free Ca2+ concentration 2-n-Butyl-4-chloro-5-hydroxymethyl-1-[(2'-(1 H tetrazol-5-yl)biphenyl-4-yl) methyl]imidazole, potassium salt (losartan), one of the antagonists, inhibited angiotensin II-induced DNA synthesis by 92% and 79%, even when added 2 and 4 h after angiotensin II stimulation, respectively. Angiotensin II also increases the mRNA of platelet-derived growth factor-A chain and basic fibroblast growth factor. The increase was observed within 4 h after angiotensin II stimulation. In this case, the addition of losartan at 4 h after angiotensin II stimulation hardly influenced the time course of the mRNA level of growth factors. Also, conditioned media of cells stimulated with angiotensin II did not influence DNA synthesis in the presence of CV11974. These results suggest that sustained receptor stimulation with angiotensin II is required for DNA synthesis in addition to the early intracellular signaling following phospholipase C activation in a manner independent of the induction of growth factors such as platelet-derived growth factor-AA and basic fibroblast growth factor. PMID- 8719429 TI - Positive and negative allosteric interactions on muscarinic receptors. AB - It was previously found that alcuronium increases the binding of [3H]methyl-N scopolamine to cardiac muscarinic receptors by a positive allosteric action while its effect on the binding of [3H]quinuclidinyl benzilate is negative. The, features of the antagonist's molecule which decide whether its allosteric interaction with alcuronium is positive or negative are not known. In the present work, it was found that alcuronium has a positive allosteric effect also on the binding of [3H]atropine and [3H]methyl-N-piperidinyl benzilate to muscarinic receptors in rat heart atria and that its effect on the binding of [3H]methyl-N quinuclidinyl benzilate is negative. A comparison of the five radiolabelled antagonists that have been investigated so far indicates that the type of allosteric interaction (positive or negative) is not determined by the presence or absence of the quaternary nitrogen or of the benzilyl moiety in the molecule of the antagonist. Apparently, features of the N-bearing moiety of muscarinic antagonists other than the presence of a charge on nitrogen play a key role in the determination of the type of interaction. PMID- 8719430 TI - GABA rho1 receptor: inhibition by protein kinase C activators. AB - The effects of protein kinase C (PKC) activators on gamma-aminoburyric acid (GABA) rho 1 receptor function were studied in rho 1 -expressing Xenopus oocytes. The PKC activator phorbol 12-myristate 13-acetate (PMA) but not the inactive analog phorbol 12-mono-myristate inhibited the GABA-gated chloride currents. Mezerein, a non-phorbol ester type PKC activator, also inhibited the rho 1 responses, but 8-chlorophenylthio-cyclic AMP, a protein kinase A activator, had no effect. The effect of PMA was significantly reduced by a PKC inhibitor, staurosporine. These results suggest that GABA rho 1 receptor function can be regulated by PKC-mediated phosphorylation events. PMID- 8719431 TI - Tranilast inhibits the effects of platelet-derived growth factor on cell proliferation and induction of nitric oxide. AB - The effects of tranilast on DNA synthesis and cell proliferation in cultured rat mesangial cells, treated with platelet-derived growth factor (PDGF), were investigated. Tranilast significantly inhibited PDGF-stimulated DNA synthesis and cell proliferation in a dose-dependent manner. In the absence of PDGF, it also enhanced cytokine-induced nitric oxide (NO) production, PDGF significantly inhibited cytokine-induced NO production, but tranilast completely abolished this inhibitory effect of PDGF. These results show that tranilast inhibits PDGF induced proliferation of mesangial cells under both normal and inflammatory conditions. PMID- 8719432 TI - Modest reduction of benzodiazepine binding in rat brain in vivo induced by antisense oligonucleotide to GABAA receptor gamma 2 subunit subtype. AB - The GABAA (gamma-aminobutyric acid-A) receptor gamma 2 subunit subtype is probably a functionally integral part of the benzodiazepine binding site of the GABAA receptor complex, important for benzodiazepine pharmacology. We have evaluated the possibility of specifically reducing benzodiazepine receptor binding properties in vivo using phosphorothioate antisense oligodeoxynucleotides to inhibit the expression of GABAA receptor gamma 2 subunit subtype. Intracerebroventricular infusions of an antisense oligonucleotide reduced benzodiazepine receptor radioligand binding by 9-15% in specific rat brain regions. PMID- 8719433 TI - The use of a computerized method of bone age assessment in clinical practice. AB - We assessed the reliability and repeatability of a new computerized bone age system, both versions 3.4 and 3.5 (licensed by Discerning System Inc. and Ares Service SA, Serono), able to automatically assess bone age on a left hand and wrist radiograph. This computer system is based upon Tanner and Whitehouse's method (TW2), but there are important differences. Our sample included an initial group of 40 patients who had growth delay/constitutional delay of growth and puberty (n = 10), growth hormone insufficiency/deficiency (n = 15), low birth weight/Silver-Russell syndrome (n = 9), precocious puberty (n = 6), as well as 20 patients with various skeletal dysplasias (multiple epiphyseal dysplasia n = 7, pseudoachondroplasia n = 7, acrodysostosis n = 5, achondroplasia n = 1), 7 girls with Turner syndrome, and 10 boys with nephrotic syndrome on chronic corticosteroid treatment. Multiple anthropometric readings of the same radiographs demonstrated excellent repeatability of the assessment. In addition, the number of times that a manual insertion of a grade was required was similar in four different assessments. The computerized method did not entirely avoid errors in interpretation as the position of the x-ray on the screen was critical. There was a high manual insertion rate in radiographs of children with skeletal dysplasia. However, the computer assessment system, version 3.5, performed adequately for radiographs of children with normal bone morphology and Turner syndrome and had the advantage of a continuous scale of assessment. PMID- 8719434 TI - Role of magnetic resonance imaging in hypothalamic-pituitary disorders. AB - Improvement of MRI diagnostic accuracy in the study of the hypothalamic-pituitary region provides precise anatomic details. In pituitary dwarfism, MRI reveals severe sella/pituitary gland and stalk hypoplasia with or without posterior pituitary ectopia, and empty sella, and this more frequently in patients with multiple pituitary hormone deficiency. Two main hypotheses have been proposed to explain these findings: traumatic stalk transection during breech delivery, and abnormal embryonic development of the pituitary gland. The association between neuroradiological findings and type/severity of endocrine alteration has not yet been clarified. In diabetes insipidus, MRI findings are normal picture, posterior lobe not visible, and thickened stalk (as expression of preclinical/initial histocytosis). Patients with central precocious puberty or hypogonadotropic hypogonadism rarely show morphologic abnormalities (hamartoma of the tuber cinereum, partially empty sella). So far, MRI permits one to identify morphologic pictures in diseases previously considered 'idiopathic'. PMID- 8719436 TI - Treatment with growth hormone. What do children know and how do they accept it? AB - Treatment with growth hormone in children is a very effective promotor of growth. The psychosocial environment of children may influence the results of this treatment. It has been shown that there is a clear relationship between the knowledge and understanding a child has of the treatment and the degree of compliance and acceptance. We have studied 90 children treated with growth hormone who filled in a questionnaire in order to know the knowledge and acceptance of their treatment. The scores for acceptance were significantly higher than for knowledge (p < 0.00001), without correlations between the two variables. The higher marks were achieved by children who had attended an educational camp. A serious lack of knowledge about the aspects directly related with the injection system was noted. We advise to design a suitable educational program for these patients. PMID- 8719435 TI - Growth hormone, growth factors and the gonad. AB - During puberty, the growth spurt and stage of pubertal maturation are two processes intimately related both being dependent on the combined action of growth hormone (GH) and sex steroids. While it is well established that both GH and sex steroids are required for a pubertal growth spurt, the precise role played by GH on the timing of onset and rate of progression through different stages of pubertal maturation is not entirely clear. A better understanding of the involved physiologic mechanisms will help clinicians in the management of adolescents with growth and/or pubertal disorders. PMID- 8719437 TI - Catch-up growth and height prognosis in early treated children with congenital hypopituitarism. AB - The aim of this retrospective study was to ascertain, whether an early growth hormone (GH) treatment can normalize height prognosis of children with congenital GH deficiency (GHD). The study covers 23 children with early onset GHD who received GH 0.1 U/kg/day from the beginning of therapy (0.4-4.9 years). This dose was corrected for weight every 3 months during the whole duration of treatment (mean 7.9 +/- 2.4 years). As a consequence of the significant growth acceleration induced by GH treatment, the patients' height deficiency at the last check had changed from a range of between -8.0 and -1.8 SDS to between +0.5 and -4.3 SDS; on the average, it was significantly less severe than before treatment. Satisfactory growth acceleration was achieved in most patients concomitantly with accelerated bone maturation, as was shown by the stable height age/bone age ratio observed during the follow-up period. The predicted ultimate height was significantly greater than the pretreatment height and it did not differ from the target height. It is concluded that catch-up growth to the target percentile in GHD patients is possible, provided that substitutive treatment is begun during the first years of life and that GH doses are adjusted periodically for weight changes. PMID- 8719438 TI - Treatment of constitutional delayed puberty with a combination of testosterone esters. AB - Thirteen boys who had constitutional delayed puberty (CDP) were treated with a combination of short- and long-acting testosterone esters (testosterone propionate, testosterone phenylpropionate, testosterone isocaproate). Mean age at the onset of treatment was 14.9 +/- 0.6 years and bone age delay was -2.7 +/- 0.9 years. An intramuscular dose of 200 mg testosterone was administered 4 times at 3 week intervals and the treated CDP boys were followed for 2 years. All boys with CDP entered puberty after the last dose (testicular volume > or = 4 ml) and growth rate increased from 4.5 +/- 0.5 cm/year pretreatment to 8.4 +/- 1.6 cm/year posttreatment after the 2-year follow-up period. Height for bone age SD score did not alter significantly from a mean of -1.1 pretreatment to -1.3 posttreatment as well as predicted height pretreatment (173.5 +/- 6.6 cm) and posttreatment (173.3 +/- 4.9 cm). A combination of testosterone esters in a given dose and schedule is a safe and effective treatment for prepubertal boys with CDP. PMID- 8719439 TI - Growth hormone therapy in Turner syndrome. Analysis of long-term results. AB - The favorable long-term results of growth hormone (GH), alone or in combination with oxandrolone, reported by an American study [J Pediatr 1992; 121:49-55], have so far not been confirmed by other investigators. However, from the presently available data it can be deduced that with higher than substitutive doses of GH, height can be improved during childhood and will be normalized in many patients with Turner syndrome. The future task must be to optimize treatment with respect to the dose and timing of GH (and oxandrolone) in individuals in order to ensure optimal statural outcome at the lowest risk and material costs. PMID- 8719440 TI - Growth hormone and dysmorphic syndromes. AB - In order to study the pathogenesis of short stature in some of the best known and most frequent dysmorphic syndromes, we have reviewed the most significant studies conducted on somatrotropin secretion and on response to replacement treatment with human growth hormone in pediatric patients. In particular, the study examines the results presented in the literature, and in a few of our cases, those obtained with regard to Noonan, Silver-Russell and Prader-Willi syndrome patients, to achondroplasia and hypochondroplasia patients, and to Down syndrome patients. Finally, we shall present a review of a few, less frequent dysmorphic syndromes with short stature, in which a growth hormone deficiency has been diagnosed and replacement treatment attempted. PMID- 8719441 TI - Use of combined Gn-RH agonist and hGH therapy for better attining the goals in precocious puberty treatment. AB - We studied 30 girls (age 6.36 +/- 1.21 years, range 4.6-8.8) affected by idiopathic precocious puberty with significant reduction of height velocity (below the 25th centile) at the end of 1 year of Gn-RHa (triptorelin intramuscular depot) treatment, to evaluate GH-IGF-I axis activity and the effects of combined Gn-RHa plus hGH therapy. After 12 months, 15 patients continued Gn-RHa and started hGH therapy for 12 months, while 15 continued treatment with Gn-RHa alone (control group). We evaluated height velocity, bone age, urinary GH, serum IGF-I and IGFBP-3 levels throughout the study; plasma GHBP levels were determined only in the first 12 months of Gn-RHa treatment. Height velocity decreased significantly during Gn-RHa treatment; it increased significantly and became higher than the control group after 12 months of Gn-RHa plus hGH treatment. During Gn-RHa therapy alone, bone age progressed less than chronological age, while in the 12 months of Gn-RHa plus hGH treatment there was a slight nonsignificant increase in bone age progression in comparison to controls. Serum IGF-I and IGFBP-3 levels decreased significantly at 12 months of Gn-RHa therapy and increased significantly after Gn-RHa plus hGH treatment. Urinary GH levels showed the same behavior. Plasma GH binding to peak II-BP, slightly lower than the prepubertal normal range before treatment, significantly increased after 12 months of Gn-RHa treatment. Therefore, in these girls, during Gn-RHa treatment alone, we have a reduction in GH-IGF-I axis activity. During Gn RHa plus hGH therapy there was a significant increase in height velocity, in urinary GH levels, in serum IGF-I and IGFBP-3 levels. Bone age did not seem to advance faster than chronological age and this may imply a better prediction in adult height. In our opinion, only in a small percentage of patients affected by precocious puberty (with a very low predicted adult height or an important reduction of growth velocity during Gn-RHa treatment) may an association with hGH therapy be useful. PMID- 8719442 TI - Effects of growth hormone on phosphocalcium homeostasis and bone metabolism. AB - In this review the effects of growth hormone (GH) on phosphocalcium homeostasis and bone metabolism are reported. Some in vitro effects of GH on chondrocytes and osteoblasts are discussed too. The main GH effects on phosphocalcium homeostasis are the permissive action on renal 1 alpha-hydroxylase activity by the hypophosphatemic stimulus and the antiphosphaturic effect by the stimulation of the maximum rate of renal tubular reabsorption of phosphate. On bone, GH is able to stimulate bone turnover and to increase bone mass. In addition, GH stimulates type I and type III collagen metabolism. In vitro, GH increases the proliferation of chondrocytes from the human growing cartilage together with the levels of interleukin-6 in the supernatant. The hormone increases also the proliferation of the human osteosarcoma-derived osteoblast-like cells and augments the osteocalcin levels in the supernatant. Thus, GH markedly influences phosphocalcium homeostasis and bone metabolism in childhood and adolescence. In addition, it is possible that GH continues to play a role in bone physiology during adulthood when final height is reached. PMID- 8719443 TI - Metabolic effects of GH: a rationale for continued GH treatment of GH-deficient adults after cessation of linear growth. AB - GH deficiency in children has been treated for decades with GH in order to normalize growth and final height. After cessation of linear growth GH treatment has traditionally been stopped. However, adults with GH deficiency have increased mortality due to cardiovascular disease and share several clinical signs, such as reduced muscle strength, reduced exercise capacity, reduced sweating ability and thermoregulation, subnormal kidney function, reduced myocardial function, reduced energy expenditure, abnormal thyroid hormone metabolism, impaired psychosocial well-being, decreased lean body mass and decreased bone mineral content. Several studies have shown unanimously that GH replacement therapy in GH-deficient adults have improved all the above-reported clinical symptoms. We conclude that GH deficiency in adults is associated with a number of subnormal features. GH treatment improves (normalizes) these symptoms and we therefore believe that GH replacement therapy of a GH-deficient individual should be continued after cessation of linear growth. However, treatment of such patients should be carefully monitored since data from follow-up studies are still lacking. PMID- 8719444 TI - Safety of growth hormone after treatment of a childhood malignancy. AB - The use of growth hormone therapy in children with radiation-induced growth hormone (GH) deficiency is widely accepted, but the safety of this mitogenic hormone, particularly in children previously treated for cancer, continues to cause concern. A variety of malignant tumours have been induced in animals exposed to supraphysiological doses of GH, whereas hypophysectomised animals appear protected from carcinogen-induced neoplasms. Growth hormone and insulin like growth factor-1 have been shown to stimulate both proliferation and transformation of normal and leukaemic human lymphocytes in vitro when used in supraphysiological doses. Despite the theoretical arguments, there is no evidence of an increased risk of tumour recurrence following GH therapy in replacement dosage in children previously treated for a malignancy. PMID- 8719445 TI - Mechanism of the antiproliferative action of leflunomide. A77 1726, the active metabolite of leflunomide, does not block T-cell receptor-mediated signal transduction but its antiproliferative effects are antagonized by pyrimidine nucleosides. AB - BACKGROUND: Leflunomide, a novel immunosuppressive drug, prolongs experimental graft survival effectively and has been well tolerated in patients with rheumatoid arthritis. A77 1726, the active metabolite of leflunomide, inhibits lymphocyte proliferation in vitro. This study was conducted in Jurkat T cells to investigate the effects of A77 1726 on signal transduction pathways initiated by ligands of the T-cell receptor CD3 complex and to evaluate the effects of A77 1726 on nucleotide biosynthesis. METHODS: Tritiated thymidine incorporation and cell counts quantitated cell proliferation. Spectrofluorescence of Indo/AM dye measured intracellular Ca2+ mobilization. A luciferase assay quantitated interleukin-2 gene promoter activity in stimulated cells transfected with an interleukin-2 promoter-luciferase gene construct. Pyrimidine and purine nucleosides were used to assess antagonism of the antiproliferative activity of A77 1726. RESULTS: (1) A77 1726 dose-dependently inhibited the proliferation of Jurkat T cells (inhibitory concentration of 50% = 6 mumol/L); (2) A77 1726 did not decrease mobilization of intracellular Ca2+ stimulated by phytohemagglutinin or anti-CD3 monoclonal antibody; (3) A77 1726 did not inhibit interleukin-2 gene promoter activity in cells stimulated with ionomycin plus phorbol myristate acetate; (4) inhibition of cell proliferation by A77 1726 was antagonized by addition of uridine, cytidine, or 2(+)-deoxycytidine; (5) addition of uridine 24 hours after treatment with A77 1726 antagonized inhibition of proliferation; (6) A77 1726 was not antagonized by 2'-deoxyuridine, thymidine, adenosine, or guanosine. CONCLUSIONS: (1) A77 1726 inhibited Jurkat T-cell proliferation without inhibiting T-cell receptor-mediated signal transduction events, including tyrosine kinase-dependent intracellular Ca2+ mobilization and activation of the interleukin-2 gene promoter; (2) the antiproliferative effects of A77 1726 on Jurkat T cells are primarily due to interruption of de novo pyrimidine nucleotide biosynthesis. These data provide evidence for a novel in vitro mechanism of the antiproliferative action of this immunosuppressant. PMID- 8719446 TI - Alloantigen processing and presentation. PMID- 8719447 TI - Only optimal donors should be accepted for heart transplantation: protagonist. PMID- 8719448 TI - Only optimal donors should be accepted for heart transplantation: antagonist. PMID- 8719449 TI - Is it necessary to perform surveillance endomyocardial biopsies in heart transplant recipients? AB - BACKGROUND: Routine surveillance endomyocardial biopsies to diagnose unsuspected rejection are performed at 3- to 12-month intervals after heart transplantation. From 1979 to 1989, surveillance biopsies were routinely performed as a part of the yearly evaluation. METHODS: A retrospective analysis of the follow-up data showed that "routine surveillance biopsies" had an extremely low yield, and, on the basis of the results of this study, we discontinued to perform surveillance biopsies beyond 6 months after transplantation. To validate these results, we compared the outcome of two groups of patients who had similar demographics and identical immunosuppression, except that in one group the surveillance biopsies were not performed. RESULTS: No difference was found in either actuarial survival rate or freedom from late rejection between the two groups. CONCLUSIONS: These findings confirm that routine surveillance heart biopsies beyond 6 months after transplantation are not necessary and they should be performed only if there is clinical suspicion of rejection or as part of a research protocol. PMID- 8719450 TI - Routine surveillance myocardial biopsies are unnecessary beyond one year after heart transplantation. AB - BACKGROUND: Myocardial rejection is most apt to occur in the first 90 days after heart transplantation. Nevertheless, surveillance endomyocardial biopsies are often performed on a regular basis, indefinitely. The benefit of this approach to patient management is uncertain. Our objective was to determine the frequency of abnormalities and the influence of a routine annual endomyocardial biopsy on patient management. METHODS: In a consecutive series of 235 transplant recipients who survived 1 year or more, the results of 1123 routine endomyocardial biopsies performed 1 year or more after transplantation were reviewed. The incidence of late rejection, presence of Quilty effect (focal endocardial or myocardial lymphocytic aggregates), and therapeutic reaction to the biopsy result were analyzed. RESULTS: Of 1123 biopsy specimens in 235 patients (1 to 12 years after transplantation), 1115 (99.3%) showed no evidence of significant rejection (grade 0 or 1). Only seven (0.6%) had evidence of rejection grade 2 or worse. Of the seven abnormal biopsy specimens in seven patients, two occurred at 1 year, two at 2 years, and one each at 4, 7, and 8 years. Of these, six were treated for rejection with an increase in the immunosuppressive therapy. One patient was identified as having a symptomatic condition at the time of biopsy. A focal endocardial or myocardial accumulation of lymphocytes (Quilty effect) was present in 311 biopsy specimens (27.6%). Beyond 1 year, 33 patients died, 14 because of graft vascular disease with or without rejection and 19 because of other causes. No deaths were predicted on the basis of a routine surveillance biopsy. CONCLUSIONS: Myocardial rejection is rare beyond 1 year after transplantation. The routine endomyocardial biopsy does not significantly impact patient management beyond 1 year. A selective approach to myocardial biopsies, on the basis of a change in clinical status or immunosuppressive medications, is justified. PMID- 8719451 TI - Endothelin plasma levels in acute graft rejection after heart transplantation. AB - BACKGROUND: Endothelin is an oligopeptide of endothelial origin with potent vasoconstrictive and mitogenic properties, implicated in the pathogenesis of cyclosporine-induced hypertension, graft vasculopathy, and renal failure. Experimental animal data suggest a role for endothelin in allograft rejection also. METHODS: To determine the role of endothelin in acute graft rejection after heart transplantation, we determined endothelin plasma levels in 165 blood samples from 79 cardiac allograft recipients (2 to 81 months after the operation) with normal graft function and correlated our findings with the histologic severity of acute graft rejection according to International Society for Heart and Lung Transplantation grading. For comparison endothelin levels were determined in 30 healthy controls and in 22 early postoperative transplant recipients (< 2 months after the operation). RESULTS: Endothelin plasma levels were significantly higher in transplant recipients than in controls (early postoperative: 7.97 = 7.53 pg/ml; late postoperative: 3.68 +/- 1.72 pg/ml; controls: 1.55 +/- 0.89 pg/ml). Endothelin plasma levels were not significantly different between groups of rejection grades 0 to 4. In the comparison of two groups of no rejection or lower (International Society for Heart and Lung Transplantation grade 0 and 1, n = 134) and higher (International Society for Heart and Lung Transplantation grade > or = 2, n = 31) rejection severity or comparing patients requiring rejection therapy (n = 20) with those not requiring therapy (n = 145), endothelin levels did not differ significantly between the groups. In 22 patients with three to six available consecutive biopsy scores and endothelin levels, intraindividual longitudinal analysis did also not show any significant correlation. The only positive correlation of endothelin levels with other laboratory parameters was found with serum creatinine concentrations (p < 0.001). In the early postoperative recipients, no correlation of endothelin plasma levels with rejection severity was seen; furthermore the only significant association was found with time after operation. CONCLUSIONS: In this study endothelin plasma levels were not influenced by acute allograft rejection after heart transplantation. Therefore endothelin levels do not appear to be a useful marker for noninvasive rejection diagnosis. Furthermore, a relevant pathogenetic role of endothelin in the rejection process cannot be derived from these data. PMID- 8719452 TI - Signal-averaged electrocardiography and Doppler echocardiographic study in predicting acute rejection in heart transplantation. AB - BACKGROUND AND METHODS: In a prospective protocol for noninvasive diagnosis of acute cardiac rejection, 83 routine endomyocardial biopsies, followed each time by the analysis of signal-averaged electrocardiography and by a cardiac Doppler echocardiographic study, were performed in 18 heart transplant recipients. The follow-up time was 5 +/- 3.6 months. To detect noninvasively acute cardiac rejection, we compared biopsy findings with the presence of late potentials at signal-averaged electrocardiography and with two diastolic indexes, pressure half time, and isovolumic relaxation time obtained from Doppler echocardiographic study. RESULTS: Thirteen acute rejection crises requiring modification of immunosuppression were diagnosed by means of endomyocardial biopsy. This clinically relevant acute cardiac rejection was associated with the presence of late potentials in 69% of cases and with the presence of pressure half-time < or = 55 msec and isovolumic relaxation time < or = 60 msec in 69% and 62% of cases, respectively. Sensitivity and specificity were as follows: for late potentials, 69% and 71%; for pressure half-time < or = 55 msec, 69% and 76%; for isovolumic relaxation time < or = 60 msec, 62% and 83%, respectively. The presence in a single patient of at least one abnormal parameter showed a sensitivity of 100% and a specificity of 60% in detecting important rejection. CONCLUSIONS: These data support the use of combined signal-averaged electrocardiography and Doppler echocardiographic study of the left ventricular diastolic function in the screening of acute cardiac rejection. Such results can suggest when endomyocardial biopsy should be performed, with the reliance that a normal noninvasive study highly excludes the presence of acute cardiac rejection requiring intensified immunosuppression. PMID- 8719453 TI - Basis for aerobic impairment in patients after heart transplantation. AB - BACKGROUND AND METHODS: To evaluate the physiologic basis for the suboptimal peak oxygen uptake observed after heart transplantation, we calculated the functional aerobic impairment ([(peak predicted oxygen uptake-peak observed oxygen uptake)/peak predicted oxygen uptake] x 100) and related it to donor/recipient, operative, and maximal exercise variables. Fifty-seven heart transplant recipients (mean age 50 +/- 10 years, 1 to 9 years after transplantation) underwent maximal upright cycle exercise testing. Concomitant exercise central hemodynamic measurements were obtained in 36 patients (63%). RESULTS: The mean peak oxygen uptake was 21.7 +/- 6.5 ml/kg/min and functional aerobic impairment was 34% +/- 17%. Functional aerobic impairment correlated positively (p < 0.01) with peak systemic vascular resistance (r = 0.55) and negatively with peak cardiac index (r = -0.62) and peak systemic arteriovenous oxygen difference (r = 0.66). A weak correlation was found between functional aerobic impairment and the duration of cardiac disease (r = 0.35, p < 0.01) but not the origin of heart failure. No correlation was seen between functional aerobic impairment and donor age, total ischemic time, time since transplantation, recipient age, and resting and exercise right and left ventricular filling pressures. CONCLUSIONS: These results suggest that the decreased exercise capacity observed in heart transplant recipients is in part due to increased peripheral vascular resistance and decreased oxygen extraction possibly due to skeletal muscle atrophy. These factors may be the result of irreversible changes from long-standing heart disease, deconditioning, or the effect of cyclosporine and prednisone. PMID- 8719454 TI - Long-term follow-up of heart transplant recipients requiring permanent pacemakers. AB - BACKGROUND: Permanent pacemaker implantation after heart transplantation is contentious. Indications for these devices in this population are uncertain. The goals of this project were to (1) analyze the time course of donor sinus node dysfunction and atrioventricular block after heart transplantation; (2) evaluate which selected parameters (donor age, ischemic time, heart rate before pacer insertion, and number of rejection episodes) might relate to persistent permanent pacing need, and (3) assess pacemaker complications during follow-up. METHODS: A retrospective analysis of pacemaker implantations (22 cases) was performed in 286 consecutive heart transplantations performed between February 1984 and April 1994 at The Methodist Hospital and Baylor College of Medicine, Houston, Texas. RESULTS: Permanent pacemakers were inserted early after transplantation in 19 patients (mean 24 days); 14 pacemakers were for sinus node dysfunction (bradycardia in five, sinus arrest with junctional escape in eight, and optimization of hemodynamics in one). Symptomatic complete heart block prompted insertion late in two patients (3 and 47 months), and symptomatic sinus pause was the indication for late insertion in one. Recipient mean age was 52.4 years, with mean donor age 29.7 years in patients with pacemakers. By 3 months, 13 of 19 patients receiving pacemakers early (mean preinsertion heart rate 58.3 beats/min) became pacer independent with subsequent mean intrinsic heart rate of 97 beats/min. Recipient or donor age, ischemic time, and rejection episodes did not appear related to long-term pacing need early or late after transplantation. CONCLUSIONS: Inferences from these observations include the fact that many patients with early sinus node dysfunction and bradycardia are not pacer dependent at 3 months. However, those with atrioventricular block early appear to require long-term pacing support. However, the possibility that more aggressive and long-term oral chronotropic medication use after transplantation would obviate early permanent pacemaker need is not addressed. Finally, prospective clinical trials are necessary to precisely characterize benefit of permanent pacemakers and define optimal pacing modes after heart transplantation. PMID- 8719456 TI - Utility of surveillance biopsies in infant heart transplant recipients. AB - BACKGROUND: Endomyocardial biopsy remains the primary means of rejection surveillance after orthotopic heart transplantation in adults. Perpetual surveillance endomyocardial biopsy has been questioned, however, because of low yield beyond the early posttransplantation period. This issue has not been adequately studied in the pediatric population. The objectives of this study were to define the rate of rejection in infants undergoing orthotopic heart transplantation, correlate rejection with signs and symptoms, and evaluate the utility of surveillance endomyocardial biopsy. METHODS: Records of all patients 24 months of age or younger undergoing orthotopic heart transplantation were reviewed; 38 patients underwent 42 transplantation; 256 endomyocardial biopsies were performed for surveillance, cardiac symptoms, noncardiac symptoms, or lowered immunosuppression. RESULTS: There were 22 rejection episodes International Society for Heart and Lung transplantation grade 1B or higher, half of which occurred in neonates 30 days of age or younger. Linearized rejection rates and actuarial freedom from rejection were not different between neonates and older infants. Linearized rejection rates reached a plateau 3 months after orthotopic heart transplantation of 0.07 episodes/100 patient days. No positive surveillance endomyocardial biopsies were obtained beyond 6 months after orthotopic heart transplantation. The probability of a positive biopsy (International Society for Heart and Lung Transplantation grade 1B or higher) was 20% or more for any other indication (odds ratios for rejection were 12.9 for cardiac symptoms, 3.3 for noncardiac symptoms, and 10.8 for lowered immunosuppression as determined by logistic regression more than 6 months after orthotopic heart transplantation). CONCLUSIONS: Rejection rates are not different between neonatal and older infants, and endomyocardial biopsies done solely for surveillance beyond 6 months after orthotopic heart transplantation rarely yield positive results. PMID- 8719455 TI - Techniques for bronchial anastomosis. AB - BACKGROUND: Many techniques have been described to optimize the construction of the bronchial anastomosis in lung transplantation. Over the past 60 months we have performed 86 bronchial anastomoses in 70 patients receiving single lung or bilateral single lung transplants. METHODS: No anastomosis was wrapped and no attempt was made at revascularization of bronchial arteries. A continuous nonabsorbable suturing technique was used in all cases. Standard triple-drug immunotherapy with cyclosporine, azathioprine, and prednisone (starting at day 7) was used for each patient. RESULTS: There were no anastomotic leaks, and seven stenoses were identified in five patients (7%). All complications were managed conservatively with stenting, and there were no related deaths. Mean time to stent placement was 109 days. One patient had bilateral stents placed prophylactically during an episode of severe infection for questionable anastomotic viability but without evidence of airway necrosis or obstruction. This patient died of infection at 16 days. Another patient died with stents in place at 71 days. In the four remaining patients, all stents have been removed after a mean of 310 days. These patients were followed up with serial bronchoscopy and were without evidence of recurrent obstruction at 2, 34, 35, and 36 months. Six of seven stenoses occurred in patients with cystic fibrosis. In each patient where stenosis developed the anastomosis was telescoped. Since abandoning the telescoping technique in the remaining 50 anastomoses (14 in patients with cystic fibrosis), no dehiscence or stenosis was encountered. CONCLUSIONS: These data suggest that elaborate techniques aimed at construction of the bronchial anastomosis are not necessary. Moreover, attempts at telescoping may be detrimental. Patients with cystic fibrosis may be a population at higher risk for anastomotic complications. Airway complications can be managed conservatively with good results and little risk to the patient. PMID- 8719457 TI - Impact of psychologic functioning, medical severity, and family functioning in pediatric heart transplantation. AB - BACKGROUND AND METHODS: This study examined (1) the psychologic functioning of children and adolescents before and after heart transplantation and (2) whether pretransplantation psychologic functioning, posttransplantation medical severity, and family functioning were related to the patients' posttransplantation psychologic functioning. The subjects were 23 patients, ages 3 to 20 years, who underwent heart transplant and survived at least 1 year after their transplantation. Psychologic functioning was assessed by the Children's Global Assessment Scale before and after heart transplantation. Medical severity was assessed by number of outpatient visits, hospitalizations, biopsies, and the Side Effect Severity Scale. Family functioning was rated on the Global Assessment of Family Relational Functioning Scale. RESULTS: The majority of the patients (78.3%) had good psychologic functioning after their heart transplantation. Patients with psychologic difficulties before and after transplantation had more hospitalizations after transplantation. CONCLUSIONS: Pretransplantation emotional functioning and family functioning were more correlated with posttransplantation psychologic functioning than medical side effect severity. PMID- 8719458 TI - Occult restrictive hemodynamics after pediatric heart transplantation. AB - BACKGROUND: Although resting hemodynamics after pediatric heart transplantation are generally within normal limits, we hypothesized that occult restrictive hemodynamics suggesting diastolic dysfunction may be unmasked by acute volume loading (fluid challenge) during cardiac catheterization. We wished to determine the incidence of diastolic dysfunction and to assess whether it progressed over time. METHODS: From 1988 through 1993, a total of 100 fluid challenges were performed at the time of surveillance endomyocardial biopsy in 31 survivors of orthotopic heart transplantation. Cyclosporine-based immunosuppression was used in 16 patients, and FK506 was used in 15 patients. Right heart hemodynamics and cardiac output (thermodilution) were obtained at baseline and after a fluid challenge with 10 ml/kg of normal saline solution. The data were analyzed to determine whether type of immunosuppression or time elapsed since transplantation predicted the response to fluid challenge. RESULTS: Baseline hemodynamics were normal; however, a marked increase in atrial filling pressures occurred after fluid challenge (p < 0.001). Findings were similar in cyclosporine- and FK506 treated patients. Hemodynamic response to fluid challenge was not related to duration of time since transplantation, including studies on patients surviving more than 4 years. CONCLUSIONS: Diastolic dysfunction after heart transplantation is common; however, the abnormalities do not progress in severity, suggesting stable long-term graft function. PMID- 8719459 TI - Short-term survivors of pediatric heart transplantation: an autopsy study of their pulmonary vascular disease. AB - BACKGROUND: The most common indications for heart transplantation in children are congenital heart disease and cardiomyopathy. Intracardiac lesions that vary widely in morphology may have a similar impact on pulmonary vascular morphology; for example, any lesion that increases left-sided intracardiac pressure will affect pulmonary venous pressure and morphology and, if long-standing, may affect pulmonary arteries also. METHODS: The lungs of eight children who died 2 days to 7 months after receiving a heart transplant at ages 8 days to 4 years were examined at autopsy with particular regard to the pulmonary arteries and veins. Arteries were evaluated for medial thickness, intimal proliferation, and peripheral extension of muscle; veins for medial thickness and mural elastic tissue configuration; and lymphatics for degree of dilation. RESULTS: The main pulmonary arteries of the children with congenital heart disease showed an abnormal elastic tissue pattern, similar to the pattern of the aorta, indicating that pulmonary arterial hypertension was present in these children at birth. In contrast, the pulmonary trunk of a child who had myocarditis beginning at age 2 years had a normal pulmonary elastic tissue pattern, consistent with normal neonatal pulmonary pressure. Five children with morphologic evidence of early pulmonary arterial hypertension had a left-to-right atrial shunt before the operation and elevated pulmonary artery pressure immediately after transplantation. The pulmonary arteries of all five patients had medial hypertrophy or peripheral extension of muscle; the most advanced pathologic condition occurred in the arteries of the oldest child, who was 5 months of age, at the time of transplantation who died 2 days after the operation with pulmonary arterial hypertension. In the lungs of all eight patients, pulmonary veins showed medial hypertrophy and excess elastic tissue fibers (arterialization), consistent with venous hypertension. The most impressive venous changes in the lungs of the children with hypoplastic left heart syndrome occurred in the two children who had the smallest left ventricles (4 x 6 mm and 8 x 8 mm), and in another child with hypoplastic and anomalous extrapulmonary pulmonary veins. The most impressive lymphangiectasis occurred in the lungs of the child with hypoplastic left heart syndrome who was oldest at the time of transplantation and in the lungs of a child with obstructed venous drainage. CONCLUSIONS: Pulmonary vasculopathy in children who require heart transplantation because of congenital heart disease or long-standing congestive heart failure may involve arteries, veins, and lymphatics. Among the eight patients in this autopsy study of children who died less than a year after heart transplantation, arterial vasculopathy associated with congenital heart disease appeared more pronounced in the five who survived less than 30 days compared with the two longer survivors. The lack of morphologic progression in short-term survivors of pulmonary arterial vasculopathy after heart transplantation correlates with clinical follow-up of long-term survivors who do not have problematic pulmonary arterial hypertension. Pulmonary venous vasculopathy is determined by the degree of pretransplantation left-sided obstruction caused by congenital or acquired disease and may be more persistent morphologically than arterial vasculopathy. This morphologic study of the pulmonary vasculature of short-term survivors of heart transplantation supports the concept that palliative surgical procedures may benefit children awaiting heart transplantation, especially infants with hypoplastic left heart syndrome for whom suitable donors are scarce. PMID- 8719460 TI - Effects of calcium concentration, lactobionate content, and sodium/ potassium ratio of preservation solutions on resting left ventricular pressure and postreperfusion function of rabbit heterotopic heart transplants. AB - BACKGROUND: We tested the hypothesis that the University of Wisconsin solution has a ionic composition (i.e., intracellular, calcium-free, lactobionate enriched) that may be beneficial for cold heart graft preservation independently from any additives. METHODS: St. Thomas' Hospital and University of Wisconsin solutions were compared with the following: (1) C solution, a simplified University of Wisconsin-like solution (i.e., intracellular, calcium-free, lactobionate-enriched); (2) A solution, an St. Thomas' Hospital-like solution (extracellular, calcium [Ca2+] = 1.2 mmol/L) in which chloride was replaced by lactobionate; (3) B solution, an intracellular, lactobionate-enriched, calcium containing solution ([Ca2+] = 1.2 mmol/L). Rabbit hearts were transplanted heterotopically in the abdomen of recipient animals either immediately or after 6 hours of storage. Hemodynamic parameters were recorded 60 minutes after unclamping. RESULTS: After a 6-hour storage, University of Wisconsin and C solutions provided better preservation than B and St. Thomas' Hospital solutions: diastolic pressures were lower; developed pressure and rate of pressure rise were higher. C solution was superior to University of Wisconsin solution only for rate of pressure rise. A solution was intermediary. A significant alteration of resting pressure and hemodynamic parameters was generally observed during the 6 hour storage. Nonsignificant changes of developed pressure and rate of pressure rise were only observed in C and B solutions: This is explained by systolic alteration after immediate reimplantation for the B group and good preservation for the C group. Resting pressure was unchanged over a 6-hour storage only for the C group, but this measure was not determined for University of Wisconsin. A correlation exists for various left ventricular volumes between resting pressure and postreperfusion hemodynamic data. Replacement of chloride by lactobionate (A versus St. Thomas' Hospital) may have improved resting and diastolic pressures by other mechanisms than limitation of net water gain during storage. PMID- 8719461 TI - Leflunomide prolongs pulmonary allograft and xenograft survival. AB - BACKGROUND: Leflunomide, an isoxazole derivative, has been shown to effectively prolong rodent allograft and cardiac xenograft survival. In vitro studies suggest that leflunomide inhibits the production of donor-specific antibodies and is capable of blocking both T- and B-cell proliferation. In light of the significant role that humoral immunity is believed to play in chronic pulmonary allograft rejection as well as hyperacute and accelerated acute xenograft rejection, we examined the efficacy of leflunomide in prolonging pulmonary allografts and xenografts and its effect on donor-specific antibody production. METHODS: Lungs from Brown Norway rats or Golden Syrian hamsters were orthotopically transplanted into Lewis rat recipients. Allograft recipients were treated daily for 14 days with vehicle, leflunomide (15 mg/kg/day orally), or cyclosporine (7.5 mg/kg/day orally) starting on the day of grafting (day 0). In xenograft recipients, leflunomide (20 mg/kg/day orally) or cyclosporine (7.5 mg/kg/day orally) treatment initiated on day 0 was continued until complete graft rejection; the leflunomide dosage was reduced to 10 mg/kg/day after day 14 because of weight loss and leukopenia. Graft viability was assessed with chest radiography in conjunction with open lung biopsies. Toxicity was monitored with body weight measurements, complete blood counts, and serum chemistries. Flow cytometric analysis of serum samples taken from graft recipients on day 7 was used to measure donor-specific immunoglobulin M and immunoglobulin G antibody titers. RESULTS: Allograft and xenograft control animals receiving vehicle yielded graft survival times of 6.0 +/- 0.0 and 5.4 +/- 0.6 days, respectively. Although xenograft recipients treated with cyclosporine (7.5 mg/kg/day orally) showed no significant graft prolongation, pulmonary allograft survival in recipients receiving cyclosporine alone was significantly prolonged to 28.2 +/- 0.7 days. Leflunomide-treated allograft (15 mg/kg/day orally) and xenograft (20 mg/kg/day orally) recipients displayed significant graft prolongation to 28.2 +/- 0.7 days and 15.8 +/- 3.3 days, respectively. Cyclosporine (7.5 mg/kg/day orally) enhanced the effect of leflunomide (20 mg/kg/day orally) in xenograft recipients with a mean graft survival time of 36.0 +/- 3.0 days achieved when both drugs were administered concomitantly. Cyclosporine significantly suppressed donor-specific immunoglobulin G antibody titers in both pulmonary allograft and xenograft recipients while not affecting immunoglobulin M levels. Leflunomide markedly suppressed both immunoglobulin G and immunoglobulin M donor-specific antibody titers in allograft and xenograft recipients. Except for mild leukopenia and anemia, both cyclosporine- and leflunomide-treated allograft recipients showed no evidence of toxic side effects after 14 days of therapy. However, leflunomide treated xenograft recipients displayed significant weight loss, anemia, and leukopenia after 14 days of treatment with one death in each treatment group. PMID- 8719462 TI - Distinct expression of cell-cell and cell-matrix adhesion molecules on endothelial cells in human heart and lung transplants. AB - BACKGROUND AND METHODS: Intercellular reactions and cell-matrix interactions are mediated by a number of specific adhesion molecules. For the intravascular reactivity of leukocytes and thrombocytes the endothelial expression of adhesion ligand molecules is of main importance. This condition may be of special relevance for the organ-specific manifestation of immune reactions in heart and lung transplants. The question was investigated as to whether organ-specific differences exist on arterial, venous, and capillary endothelial lining cells in heart and lung transplants and whether this condition is modified during transplant rejection. Transplant biopsy specimens of 24 heart transplant recipients (n = 303) and lung transplant recipients who underwent retransplantation for rejection (n = 4), as well as normal heart (n = 7) and lung tissue (n = 4), were studied. Cell-cell adhesion molecules (intercellular adhesion molecule-1 (CD54) and -2; LFA-3; CD31; vascular cellular adhesion molecule-1; neural cellular adhesion molecule; E-/P-selectin; CD44) and cell matrix adhesion molecules (very late antigen-1 through -6; CD51) were studied on cryostat sections by means of standard immunohistology. RESULTS: (1) Arterial, venous, and capilary endothelia in lung and heart tissue show a distinct pattern of cell-cell and cell-matrix adhesion molecules: capillary and venous endothelia are in contrast to arterioles negative for vascular cellular adhesion molecule-1 and P-selectin. Arterial endothelia of the lung, in contrast to veins and capillaries, express no receptors for laminin (very late antigen-2 and very late antigen-6). (2) With transplant rejection, an induction of a number of adhesion molecules was noted on all endothelial lining cells in heart and lung transplants (intercellular adhesion molecule-1[CD54], intercellular adhesion molecule-2, lymphocyte function antigen-3, very late antigen-2, and very late antigen-6). Capillaries of the lung, in contrast to heart capillaries, displayed no inducibility of vascular cellular adhesion molecule-1 and E-selectin. In heart transplants, differences between capillaries and arteriovenous endothelia were found for virus-like agent-2 and -3. CONCLUSIONS: These results show a distinct expression of a both cell-cell and cell-matrix adhesion molecules on arterial, venous, and capillary endothelia of heart and lung transplants. This expression may influence the local regulation of leukocyte and thrombocyte adhesion during transplant rejection. Especially lung capillaries show a lack of inducibility for vascular cellular adhesion molecule-1 and E- and P-selectin required for the induction phase of leukocyte adhesion. This lack was not found in heart transplants. This observation may explain differences in intravascular adhesion and infiltration between vessel compartments in hearts and lungs and could be of relevance for therapeutic interventions to modify leukocyte and thrombocyte adhesion, as during transplant rejection and reperfusion damage. PMID- 8719463 TI - Additive value of immunologic monitoring to histologic grading of heart allograft biopsy specimens: implications for therapy. AB - BACKGROUND: Currently the sole method available for diagnosis of heart allograft rejection is endomyocardial biopsy. Although this procedure offers important criteria for treatment, it cannot always discriminate between mild episodes of rejection which might be self-limiting and forms which may progress. In an effort to monitor rejection, we have implemented a cellular monitoring strategy aimed at identifying episodes of rejection in biopsy specimens which may evolve into higher grades of rejection. The lymphocyte growth assay is based on the capacity of interleukin-2 receptor-positive T cells to expand in the presence of interleukin-2 and antigen provided by the biopsy fragment. In this study we investigated whether a positive lymphocyte growth assay correlated with and was predictive of subsequent histologic allograft rejection and the development of anti-human leukocyte antigen antibodies. METHODS: Lymphocyte growth assay was performed on 437 biopsy specimens from 76 patients. Patients with mild allograft rejection defined as grade 2 rejection were randomized to treatment according to the results of the lymphocyte growth assay. Anti-human leukocyte antigen antibodies was also measured monthly. Cells grown from the biopsy specimens were tested against the donor cells and allopeptides derived from the donor human leukocyte antigen-DR. RESULTS: A highly significant correlation was observed between the histologic grade of rejection and growth of graft infiltrating cells (p < 0.0001). Lymphocyte growth occurred in 10% of grade 0 versus 60% of grade 3A biopsy specimens. Only 4% of histologically negative cases with negative lymphocyte growth assay progressed to rejection in the next month. In the randomized study in which treatment was based on the lymphocyte growth assay results, progressive rejection occurred in three of four cases with positive lymphocyte growth assay versus only 1 of 11 with a negative lymphocyte growth assay (p < 0.001). A highly significant correlation was found between a positive lymphocyte growth assay and subsequent development of antihuman leukocyte antigen antibodies (p < 0.0006). This finding indicates that cellular rejection evidenced by lymphocyte growth assay ultimately results in humoral antihuman leukocyte antigen antibody mediated rejection. Limiting dilution analysis showed that although the direct recognition pathway prevails in early rejection, cells participating in the indirect pathway also proliferate vigorously in the graft during rejection. CONCLUSIONS: Monitoring of rejection with lymphocyte growth assay is a simple method which provides prognostic information on the outcome of cardiac allografts. Lymphocyte growth assay correlates with histologic rejection and is predictive of future histologic rejection episodes. Lymphocyte growth assay also predicts subsequent development of antihuman leukocyte antigen antibodies and thus may provide a useful method for ascertaining the onset of chronic rejection. PMID- 8719465 TI - Audit of referral and explant diagnoses in lung transplantation: a pathologic study of lungs removed for parenchymal disease. AB - BACKGROUND: Lung transplantation is performed for an increasing range of pulmonary conditions in which the diagnosis is often clinical or based on limited biopsy material. Diagnosis may be made late in the course of the disease where specific features are no longer present. Posttransplantation complications and disease recurrence may relate to the primary disease, and accurate diagnosis is therefore essential. METHODS AND RESULTS: A pathologic review of 183 explanted lungs over a 10-year period (heart-lung = 109, single lung = 65, double lung = 9) showed 29 significant discrepancies or additional features likely to effect outcome. The final pathologic diagnosis was cystic fibrosis (n = 66), emphysema (59), bronchiectasis (17), pulmonary fibrosis (19), sarcoidosis (10), Langerhans cell histiocytosis (3), pulmonary veno-occlusive disease (3), posttransplantation obliterative bronchiolitis (2), primary hemosiderosis (1), rheumatoid obliterative bronchiolitis (1), extrinsic allergic alveolitis (1), pneumoconiosis (1). Unsuspected diagnoses included tuberculosis (8) (four cases of which were active and in single lung recipients requiring antituberculous chemotherapy), sarcoidosis (9), (of which, six were unsuspected primary diagnoses and three were additional diagnoses), veno-occlusive disease (3), carcinoma (1), pneumoconiosis (1), and pulmonary fibrosis (2). Aspergillus infection (2) and bronchocentric granulomatosis (3) were found in patients with cystic fibrosis. One active tuberculosis case also showed an aspergilloma. Unsuspected infections requiring therapy in immunosuppressed patients and previously unsuspected sarcoidosis, which is known to recur in the graft, were the major novel diagnoses. Discrepancy rate was 12 of 65 in single lungs (19%) and 17 of 109 in heart-lungs (16%). CONCLUSIONS: These results emphasize the need for accurate preoperative diagnosis especially when the similarly diseased native lung remains in situ. PMID- 8719464 TI - Nebulized cyclosporine for prevention of acute pulmonary allograft rejection in the rat: pharmacokinetic and histologic study. AB - BACKGROUND: With regard to limiting the systemic effects of cyclosporine A and obtaining better control of acute pulmonary allograft rejection, local immunosuppressive therapy with aerosolized cyclosporine A seems of interest. Given the in situ immunologic mechanisms of acute rejection, as well as the anatomic structure of the lung, this therapy is feasible as previously described by others. The aim of our study is to determine the pharmacokinetic parameters of nebulized cyclosporine A and the best modalities of administration. METHODS: In a pharmacokinetic study, the cyclosporine A was given either by intramuscular injection (10 mg/kg) or by aerosol at 10 and 25 mg/kg doses; 70 rats were killed at 25 and 50 minutes and 2, 4, 6, 8, 12, 24, or 48 hours after cyclosporine A administration. Cyclosporine A levels were measured in whole blood and in the lung. The areas under the concentration time curves were determined. Twenty-four lung transplantations were then performed. The rats were killed on postoperative day 9. Acute rejection was scored on a scale of 0 to 4, and cyclosporine A trough levels were measured in the lung and in the blood. RESULTS: With a jet nebulizer, the mass median aerodynamic diameter was 2.5 microns, with a standard geometric deviation of 2.3. In blood, the area under the concentration curve was greater for intramuscular (80.6 ng.hr/ml) than for aerosol administrations at 10 (15.1 ng.hr/ml) and 25 mg/kg (41.0 ng.hr/ml) doses. In the lungs, the area under the concentration curve was greater for the aerosol route at 25 mg/kg doses (588 ng.hr/mg) than for the low-dose (200 ng.hr/mg) or intramuscular administration (200 ng.hr/mg). The lung targeting index of cyclosporine A (ratio area under the concentration curve-lungs/area under the concentration curve-blood) was greater for both aerosol administrations than for the intramuscular route. In the study of the prevention of acute rejection, rats without immunosuppression (n = 6), rats receiving daily doses of cyclosporine A intramuscularly (10 mg/kg), and rats with aerosolized cyclosporine A daily (10 and 25 mg/kg/day) showed mean grades of acute rejection of, respectively, 4, 2.03 +/- 0.27, 2.33 +/- 0.52, and 2.17 +/- 0.46. The deposition of nebulized cyclosporine A was lower in transplanted than in native lung. CONCLUSIONS: Nebulized cyclosporine A allows better pulmonary concentration than intramuscular administration, and results in lower systemic levels. Prevention of acute rejection is as good with aerosolized cyclosporine A as with intramuscular cyclosporine A. This first pharmacokinetic study of nebulized cyclosporine A could lead to clinical applications. PMID- 8719466 TI - Does cyclosporin A have any effect on accelerated atherosclerosis in absence of graft rejection? Pathologic and morphometric evaluation in an experimental model. AB - BACKGROUND: The effects of cyclosporin A on accelerated atherosclerosis were studied in an experimental model of aortic isotransplantation. METHODS: Seventy six Lewis rats were studied. Forty-one abdominal aortic isografts were performed and divided into five groups: 2-day isografts and 15- and 100-day isografts with and without cyclosporin treatment. The remaining rats were divided into seven groups: 15- and 100-day sham-operated, with and without cyclosporin administration; 15- and 100-day animals with cyclosporin treatment only; and normal controls. Cyclosporin was injected subcutaneously in doses of 10 mg/kg daily for the first 15 days and afterward every other day. Longitudinal sections of the proximal anastomosis and cross sections of the midgraft region were measured with a semiautomatic image-analyzer. RESULTS: Histologic analysis showed that accelerated atherosclerosis was not observed either in NT2 rats or in nontransplanted animals. In the 15-day isografts, accelerated atherosclerosis was present in the perianastomotic tract of the recipient aorta in nine of nine NT15 rats, whereas it was found only in three of nine T15 animals (p < 0.02). Histomorphometric analysis showed that accelerated atherosclerosis was less pronounced in the T100 isografts than in the NT100 ones, this difference being significant at the recipient anastomotic side only (p < 0.0005). CONCLUSIONS: The present results support the hypothesis that cyclosporin, at immunosuppressant and nontoxic doses, can delay the onset and progression of accelerated atherosclerosis and that its effects are more significant at the recipient side of the anastomosis where accelerated atherosclerosis begins to develop. PMID- 8719467 TI - Pathogenesis of Quilty lesion in cardiac allografts: relationship to reduced endocardial cyclosporine A. AB - BACKGROUND: Endocardial lymphocytic infiltrates, known as Quilty effect, are a common finding of uncertain pathogenesis in cardiac allografts. Quilty effect was not observed before the use of cyclosporine A for immunosuppression and is not generally regarded as a manifestation of rejection. We hypothesized that the endocardial localization of Quilty effect may be related to a relative absence of cyclosporine A in this region. METHODS: We used an indirect immunofluorescence staining method with rabbit polyclonal anti-cyclosporine A antibodies to detect cyclosporine A in fresh frozen sections of 27 cardiac allograft endomyocardial biopsies. Staining was graded 0 to +3. Negative controls were from untreated transplant candidates and from biopsies with the primary antibody omitted. RESULTS: On comparison of endocardial and myocardial fluorescence in biopsy specimens from patients treated with cyclosporine A, there was less endocardial (0.7 +/- 1.1, p < 0.0001) than myocardial (2.2 +/- 0.5) staining. However, in biopsy specimens with Quilty effect (n = 12), this difference was significantly greater (endocardial = 0.2 +/- 0.6 versus myocardial = 2.3 +/- 0.5; p = 0.005) than in specimens without Quilty effect (n = 10) (endocardial = 1.4 +/- 1.2 versus myocardial = 2.1 +/- 0.6; p = 0.7). Endocardial thickness as measured by ocular micrometry was significantly greater in regions with (32 +/- 19 microns) than without (7 +/- 4 microns) Quilty effect, with involved regions showing increased connective tissue (p < 0.0001). In patients with and without Quilty effect, no differences in donor or recipient demographics, prevalence of diabetes, or plasma cyclosporine A levels were found. CONCLUSIONS: Although it has been postulated that Quilty effect is due to the presence of cyclosporine A in cardiac tissue (toxic effect or immunologic reaction), these data suggest that Quilty effect is related to reduced endocardial presence of cyclosporine A, leading to localized, contained, and usually not clinically significant endocardial rejection. PMID- 8719468 TI - Prognosis of alternative therapies in patients with heart failure not accepted for heart transplantation. AB - BACKGROUND AND METHODS: This article describes the outcome of alternative therapies in patients with end-stage heart failure, New York Heart Association class III-IV, referred for heart transplantation evaluation but not accepted for the procedure. From January 1988 through September 1992, 233 consecutive patients with severe heart failure were admitted to the thoracic transplantation center at Sahlgrenska University Hospital. At the time of admission all patients received standard medical treatment for heart failure. During the pretransplantation evaluation, an attempt to optimize the medical therapy was made in all patients. RESULTS: Eighteen patients (8%) died before a decision concerning transplantation was made, and 146 patients (63%) were accepted for heart transplantation. There were 69 patients (30%) who were denied heart transplantation for various reasons, and they were subgrouped: patients with contraindications (group 1, n = 23) or without indication (group 2, n = 10) for heart transplantation, patients with a positive response to intensified medical therapy (group 3, n = 25), and patients who underwent coronary artery bypass grafting and/or valvular heart surgery (group 4, n = 11). The 1-, 3-, and 5-year actuarial survival rates were as follows: group 1, 26%, 16%, and 8%; group 2, 100%, 77%, and 39%; group 3, 96%, 67%, and 53%; and group 4, 64%, 36%, and 27%, respectively. The corresponding figures for patients who had a heart transplantation were 85%, 79%, and 75%, respectively. During the first 2 to 3 years of follow-up the survival of group 2 and group 3 patients was similar to that of patients who underwent transplantation. However, late survival was worse compared with the heart transplant group. CONCLUSIONS: These results suggest that by close follow-up it may be possible to postpone heart transplantation in a selected group of patients. PMID- 8719470 TI - Successful dynamic cardiomyoplasty after heart transplantation. PMID- 8719471 TI - Allograft Coronary Disease: A Basic Science and Clinical Review. Proceedings of the 2nd International Symposium. St. Louis, Missouri, January 31-February 1, 1996. PMID- 8719469 TI - Safety and efficacy of beta blockade in patients with chronic congestive heart failure awaiting transplantation. AB - BACKGROUND: Donor availability remains a limiting factor for heart transplantation while transplant waiting time entails significant morbidity and mortality. This study was designed to assess the efficacy and safety of long-term beta blockade as optimization of therapy in patients with severe congestive heart failure already receiving digoxin, diuretics, and converting enzyme inhibitors awaiting transplantation. METHODS: The beta-1 antagonist metoprolol was given to 19 patients with moderate to severe congestive heart failure. Hemodynamic, clinical, and neurohormonal measurements were obtained before drug therapy and after 3 months of treatment. Patients initially received 6.25 mg of metoprolol orally twice daily which was increased to a target dose of 50 mg twice daily over several weeks. RESULTS: Metoprolol produced significant clinical, exercise, and hemodynamic benefits. Long-term therapy was associated with improvements in New York Heart Association class, ejection fraction, 6-minute walk, and peak maximal oxygen consumption. There were significant decreases in heart rate, pulmonary arterial systolic pressure, and left ventricular filling pressure with significant increases in stroke volume index and stroke work index. Four patients were removed from the transplant list after improving to New York Heart Association I. Only one patient required hospitalization during the first 6 months of therapy. There were no deaths caused by progressive heart failure; however, one patient died suddenly. CONCLUSIONS: Beta blockade with metoprolol can be safely administered to patients awaiting heart transplantation producing clinical, exercise, and hemodynamic improvements. Thus, beta blockade may prove to be a safe and cost-effective bridge to transplantation. PMID- 8719472 TI - Macrophages and transplant arteriosclerosis: known and novel molecules. PMID- 8719473 TI - Cytokine-mediated fibronectin production and transendothelial migration of lymphocytes in the mechanism of cardiac allograft vascular disease: efficacy of novel therapeutic approaches. PMID- 8719475 TI - Immune and nonimmune mechanisms of chronic rejection of kidney allografts. PMID- 8719474 TI - Transplant arterial vasculopathy: evidence for a dual pattern of endothelial injury and the source of smooth muscle cells in lesions of intimal hyperplasia. AB - With use of this straight artery model of transplantation for studying TGVD, we have shown the following: 1. Intimal thickening occurs in a time-dependent predictable manner only in allografts. 2. Endothelial injury as a result of procurement, preservation, and reperfusion by itself does not result in the development of intimal thickening. Additional endothelial injury associated with the presence of an early mononuclear infiltrate is necessary for the development of TGVD. 3. The development of intimal hyperplasia is initially cellular, consisting mainly of macrophages and a smaller proportion of lymphocytes. The macrophages are later replaced by SMCs. These findings are summarized in Figure 16. 4. The source of the SMCs in intimal lesions is most likely the recipient media. PMID- 8719476 TI - Cardiac allograft vasculopathy: the central pathogenetic role of ischemia-induced endothelial cell injury. PMID- 8719477 TI - The pathogenesis of graft vascular disease: implications of vascular remodeling. PMID- 8719479 TI - Biologic modification of the immune response in mouse cardiac isografts using gene transfer. PMID- 8719478 TI - Hemostasis, fibrinolysis, and natural anticoagulation in transplant vascular sclerosis. PMID- 8719480 TI - Estrogen and prevention of transplant atherosclerosis. PMID- 8719481 TI - Acute rejection: culprit or coincidence in the pathogenesis of cardiac graft vascular disease? PMID- 8719482 TI - The role of histoincompatibility in cardiac allograft vasculopathy. PMID- 8719483 TI - Cellular and humoral immunity to vascular endothelium and the development of cardiac allograft vasculopathy. PMID- 8719484 TI - Characterization of endothelial antigens associated with transplant-associated coronary artery disease. PMID- 8719485 TI - Quantitative analysis of transplant coronary artery disease with use of intracoronary ultrasound. PMID- 8719486 TI - Prevalence and distribution of transplant coronary artery disease: insights from intravascular ultrasound imaging. PMID- 8719487 TI - The prognostic significance of intimal proliferation in cardiac allograft vasculopathy: a paradigm shift. PMID- 8719488 TI - Does intracoronary ultrasound conflict with pathology? Comments from a panel discussion. PMID- 8719489 TI - Incidence and progression of transplant coronary artery disease over 1 year: results of a multicenter trial with use of intravascular ultrasound. Multicenter Intravascular Ultrasound Transplant Study Group. PMID- 8719490 TI - Does acute rejection correlate with the development of transplant coronary artery disease? A multicenter study using intravascular ultrasound. Sandoz/CVIS Investigators. PMID- 8719491 TI - The impact of cyclosporine dose and level on the development and progression of allograft coronary disease. Sandoz/CVIS Investigators. PMID- 8719492 TI - Role of lipids in allograft vascular disease: a multicenter study of intimal thickening detected by intravascular ultrasound. PMID- 8719493 TI - The role of nonimmune risk factors in the development and progression of graft arteriosclerosis: preliminary insights from a multicenter intravascular ultrasound study. Multicenter Intravascular Ultrasound Transplant Study Group. PMID- 8719494 TI - Cardiac allograft vasculopathy: the potential role for transmyocardial laser revascularization. PMID- 8719495 TI - Impact of converting enzyme inhibitors and calcium entry blockers on cardiac allograft vasculopathy: from bench to bedside. PMID- 8719496 TI - Fish oil and antioxidants after heart transplantation: future strategies or eye of newt and wing of bat revisited? PMID- 8719497 TI - Primary (idiopathic) generalized epilepsy and underlying mechanisms. AB - Primary Generalized Epilepsy (PGE) has been more hotly debated over the past decades than other forms of epileptic seizure disorder. The sudden synchronous appearance of bilateral spikes and spike-waves (mainly with myoclonus resp. absence) used to perplex the earliest generation of electroencephalographers, and the enigmatic genesis of these discharges (and seizures) has not ceased to fascinate the investigators of this phenomenon. A "centrencephalic" concept with paroxysmal discharges arising from thalamic structures and "projecting" to the cortex was championed for many years and eventually laid aside. More recently, the role of the thalamic level has been re-emphasized, mainly on the basis of experimental work. In this article, the bulk of experimental work is critically reviewed: the simian model (Papio papio), the feline, and the rodent models (Wistar rat, tottering mouse). Stress is being laid on fundamental differences between all of these models and human PGE. EEG evidence indicates a superior frontal origin of bilateral-synchronous spikes and spike-waves; depth EEG recordings in patients have failed to demonstrate primary thalamic spike generation. The heart of the matter in PGE appears to be the mechanism underlying paroxysmal discharges; above all the role of arousal. It is not awakening from sleep but the ensuing period that is critical in its epileptogenic thrust caused by alternating periods of return to drowsiness and arousing stimuli. This biphasic process gradually escalates EEG bursts to myoclonus (or absences) and possibly to a generalized tonic-clonic convulsion. Most conducive to this crescendo is the state of tiredness following a night of poor sleep. Bilateral synchrony is not precise and small time differences exist. The line between primary and secondary bilateral synchrony (with a primary cortical focus) can become blurred. Genetic predisposition to generalized paroxysms must always be considered, even in the face of a primary focus with secondary bilateral synchrony. Photosensitivity is a second paroxysm-inducing mechanism in PGE; it is much less common than the abnormal arousal ("dyshormia"); both mechanisms can be present in the same patient. Therapy and prevention of seizures in PGE are finally discussed. The concept of abnormal arousal mechanisms can be put into practice in order to prevent seizures: avoidance of sleepless nights, not always an easy task in adolescents and young adults. PMID- 8719498 TI - Dyshormia revisited: generalized seizures and arousal. AB - Niedermeyer has described dyshormia, or deviant arousal, and related this EEG pattern to generalized seizures. We describe a patient with generalized seizures who in the same EEG had spindles transforming into generalized epileptiform discharges and generalized epileptiform discharges transforming into spindles. This suggests that similar neural circuitry may underlie both spindles and generalized epileptiform discharges, and that the mechanism described by Gloor for feline generalized penicillin induced epilepsy may also apply to some humans with generalized epilepsies. PMID- 8719499 TI - Sensitivity and specificity of QEEG in children with attention deficit or specific developmental learning disorders. AB - The sensitivity and specificity of QEEG-based discriminant functions were evaluated in populations of children diagnosed with specific developmental learning disorders and those with attention deficit disorders. Both populations of children could be distinguished from each other, and from the normal population, with high levels of accuracy. Pretreatment QEEG could be utilized to distinguish ADD/ADHD children who responded to dextroamphetamine from those who responded to methylphenidate, again with high levels of accuracy. This paper provides a replication of all presented discriminant functions, and should provide the research basis for the generalized utilization of QEEG in the initial evaluation of children with learning and/or attention disorders. PMID- 8719500 TI - A review of the usefulness of the standard EEG in psychiatry. AB - The goal of this paper is to draw conclusions about the usefulness of the standard EEG in psychiatry. In general, two thirds of psychiatric referrals for an EEG are expected to provide useful information. The emphasis in schizophrenia is placed on left-sided abnormalities, especially on the left temporal area. In mood disorders the emphasis is on right-sided foci, in addition to the controversial 6/sec spike and wave complexes, small sharp spikes and positive spikes. In the acute stage of alcoholism, a relationship is seen between the degree of intoxication and the amount of slow activity, while in the chronic stage an increase in slow activity is seen, but another change is fast activity on the temporal areas. During withdrawal a low seizure threshold can be seen as irregular bilateral spike and wave complexes. During abstinence 2-4 yr may be required before slow wave sleep is normal in all regards. Among the organic mental syndromes, delirium shows slow activity, except in delirium tremens, which often is associated with a normal record with fast activity. In dementia the prevalence of EEG abnormalities is related to the degree of impairment. After five sessions of ECT diffuse slow waves are often seen. In other conditions, among developmental disorders about one half of autistic children show abnormalities and epileptiform activity is not uncommon. Mild nonspecific abnormalities are seen in about 40% of dyslexics and also in behavior disorders. Anxiety disorders include anorexia nervosa, showing abnormal background activity related to the effect of starvation on cerebral metabolism. In panic attacks paroxysmal activity can be seen. In borderline personality positive spikes have been (again) associated with impulsivity and 6/sec spike and wave complexes with interpersonal problems. Of the drugs of abuse psilocybin and phencyclidine are often associated with generalized epileptiform patterns and with marijuana the alpha shows a decreased frequency with increased amplitude. Typically, an increase in slow activity is seen with psychotropic drugs if there is a change in the level of awareness. Finally, distinctive personality traits are, at times, seen in temporal lobe epilepsy and the phenomenon of "forced normalization" may appear when seizures stop and psychotic symptoms appear. PMID- 8719501 TI - Quantitative pharmaco-EEG of carbamazepine in volunteers and epileptics. AB - In order to measure the effect of carbamazepine (CBZ) on quantitative EEG (qEEG), one dose of CBZ (300 mg) was administered to 10 epileptics and 10 healthy adult volunteers. EEG samples were recorded before and at regular intervals within 24 hours following administration. The EEG activity was processed by power spectral analysis and the drug serum concentration was assessed concomitantly. The results showed that with the increase of serum concentration of CBZ, the percentage of power progressively increased in the theta band while it decreased in the alpha 2 band. The percentage of theta and alpha 2 power was found to reach maximum and minimum levels, respectively, when the peak values of serum concentration of CBZ were attained. The effects on qEEG in anterior scalp areas were more evident than in posterior scalp areas. PMID- 8719502 TI - Topography of auditory and visual P300 in normal children. AB - Auditory and visual P300 recordings were performed on 39 normal, right-handed individuals from age 6 through 15, using 31 evenly spaced scalp electrodes. Amplitude at the P300 peak and latency to this peak at each electrode site were measured. Age was significantly correlated with the 31-electrode mean for auditory and visual P300 latencies, but not for amplitudes. The younger age group (6-10) had longer auditory and visual P300 latencies than the older age group. Visual P300 amplitudes were of an overall larger magnitude than auditory amplitudes. There were no other differences including significant topographical differences in P300 amplitudes or latencies by gender, age group, modality, or side of scalp. Radial current density maps on group-averaged auditory and visual P300 waveforms at the group mean P300 latency at Cz, showed a right centroparietal sink surrounded by sources. This suggests a major right centroparietal P300 generator. Description of the normal topography of the P300, and demonstration of the lack of topographic differences by gender, age group, modality, or side of scalp, may facilitate the meaningful examination of P300 topography in cognitive disorders. Such an examination might lead to better diagnostic tools and more appropriate treatment of cognitive disorders in children. PMID- 8719503 TI - Hypersomnia associated with a focal pontine lesion. AB - A 50-year-old woman developed intractable excessive sleepiness after undergoing the surgical removal of a brainstem cholesteatoma. The 24-hour ambulatory monitoring revealed a normal architecture of sleep contents, with 62.7% of the time spent in sleep. Auditory and somatosensory evoked responses showed abnormal patterns. The MRI scan of her brain showed an extensive nonprogressive lesion in the brainstem. We speculate that the problem underlying the patient's hypersomnia is a defect in the ascending reticular activating system (ARAS) rather than in the REM and NREM sleep mechanisms. PMID- 8719504 TI - Comparison of horizontal head movements evoked by auditory and visual targets. AB - Head movement propensity-the pattern of head saccades dependent on methods of target presentation-varies among individuals. The present group of 9 young adults was previously ranked in a visual saccadic task according to this propensity. The present report examines how and why this propensity changes if the saccades are made to auditory targets. 1) Spatially identical, interleaved, auditorily and visually elicited horizontal saccadic gaze shifts (jumps) differed in amplitude and in starting and/or ending position. The jumps were executed in two head movement modes: first, the non-aligned mode was a standard reaction-time single gaze step between two points. Second, the head-aligned mode required alignment of the head with the fixation (starting) point; thereafter both modes were identical. All results in the auditory task are expressed relative to the visual results. 2) In the non-aligned mode, head movement amplitudes were increased on average by 15% (for example, an 80 degrees jump elicited a 12 degrees larger head movement), and velocity decreased by 12%, reflecting the increased demands of the auditory task. More importantly, the differences between subjects was narrowed; that is, head movement propensity was homogenized in the auditory task. In the visual task, head-movers willingly move their heads off and across the midline, whereas non-movers are unwilling to leave the midline from eccentric starting points or to eccentric ending points. This is called the midline attraction effect and was previously linked to spatial reference frames. The homogenization in the auditory task was characterized by head-movers increasing, and non-movers decreasing, their midline attraction, suggesting altered spatial reference frames. 3) For heuristic purposes, the ideal head-mover is defined by a gain of 1.0 in the visual task, and by external earth-fixed reference frames. Similarly, the ideal non-mover has a gain of 0.0 and has a bias toward body (or some par of the body)-fixed reference frames. In the auditory task these gains (and reference frames) in head movers and non-movers are homogenized (close to 0.5), either by the participation of the head (movement of the ears in space) in sensory acquisition or by differences in central nervous processing of the two modalities, or both. PMID- 8719505 TI - Changes in the gastrointestinal electric pattern to motion sickness in susceptibles and insusceptibles during fasting. AB - The cardinal symptoms of motion sickness occur in the gastrointestinal system. This study sought to investigate the influence of sickness-provoking motion stimuli on the interdigestive migrating electric complex (IDMEC) of the gastro intestinal system. Thirteen susceptibles and 14 insusceptibles to motion sickness were subjected to intralumenal recording before and after caloric vestibular stimulations. Motion sickness developed in all subjects. The results indicate that there are no significant differences in the IDMECs of susceptibles and insusceptibles in a state of rest; vestibular stimulation changes the duration of IDMEC phases (P < 0.05 and P < 0.01); IDMEC changes differ slightly between susceptibles and insusceptibles; motion stimuli induce simultaneous nonpropulsive spike activity, which is more frequent in susceptibles (P < 0.01); vestibular stimulation applied during phase III of IDMEC does not provoke vomiting in susceptibles. PMID- 8719506 TI - Effects of long duration centrifugation on head movements and a psychomotor task. AB - Previous investigations suggest strong similarities between aftereffects of an 1.5-hour 3 gx centrifuge run (Sickness Induced by Centrifugation; SIC) and the Space Adaptation Syndrome (SAS). Head movements proved to be especially nauseogenic in both cases. Until recently, during SIC experimentation, the performance of head movements was not adequately operationalized, which hampered a statistical evaluation. For this purpose, a test was designed by which head movements in four directions (left, right, up, and down) were provoked and registered, once before and once after a 1-h 3 g centrifuge run. Subjects had to execute these head movements in order to perform well on a psychomotor task. The results indicate that the test described is sensitive in discriminating between subjects who are seriously affected after a long duration centrifugation and those who are not. Because differences in performance were found only on those parts of the task in which head movements were involved, objective evidence is obtained in favour of a vestibular cause in the etiology of SIC. PMID- 8719507 TI - Vertigo after sailing a nineteenth century ship. AB - Few papers describe illusions of motion after sailing for which subjects do not seek medical assistance. After sailing on a square-rigged ship for five-hour periods subjects were surveyed about the incidence, intensity, and quality of such illusions. Even on a calm day, 20% to 41% of the crew experienced post sailing vertigo, but it only occurred among the nonprofessional sailors. This phenomenon is characterized by extinction with repeated exposure, a lag time to onset of about two hours, brief duration, and occurrence in enclosed visual surrounds. These data are similar to vestibular habituation and suggest that some central vestibular mechanisms may be involved. PMID- 8719508 TI - Dizziness of suspected cervical origin distinguished by posturographic assessment of human postural dynamics. AB - Useful clinical tests are lacking for the controversial entity "cervical vertigo". In earlier studies patients assumed to suffer from cervical vertigo or dizziness manifested disturbed postural control as compared to healthy subjects, but were hard to distinguish from patients with other balance disorders. Using posturography in which stance was perturbed by a vibratory stimulus applied towards the calf muscles, we studied 16 consecutive patients with recent onset of neck pain and concomitant complaints of vertigo or dizziness, but normal findings at otoneurological examination and electronystagmography; 18 patients with recent vestibular neuritis; and 17 healthy subjects. We performed system identification of a model of the control of upright human stance, using the vibratory stimulus as input and the recorded body sway as output. According to values for the three normalized parameters of the transfer function of the model (i.e., swiftness, stiffness, and damping), cervical vertigo patients were distinguished both from healthy subjects (P < 0.001), and from vestibular neuritis patients (P < 0.001). It was also possible to distinguish the vestibular neuritis group from the group of healthy subjects (P < 0.01). The results show disturbed postural control in patients with cervical vertigo to differ from that in patients with recent vestibular neuritis, and indicate posturographic assessment of human posture dynamics to be a possible future tool for use in diagnosing cervical vertigo. PMID- 8719509 TI - Fentazin anesthesia for labyrinthine surgery in guinea pigs. AB - We describe the use of fentazin anesthesia (0.4 mg/ml fentanyl citrate, 58.3 mg/mL xylazine HCl, and 3.2 mg/mL azaperone, Parnell, New Zealand) for labyrinthine surgery in guinea pigs. Fentazin provides stable surgical anesthesia for approximately 1 hour, rapid recovery from anesthesia following the surgery, and has significant analgesic and anxiolytic properties. Its advantages in comparison to other anesthetics (for example, ketamine HCl) are discussed. PMID- 8719511 TI - Vestibular bibliography. PMID- 8719510 TI - Influence of arousal and attention on the control of postural sway. AB - Healthy young adults (n = 39) were asked to perform four different secondary cognitive tasks during quiet unperturbed stance, in order to investigate the influence of physiological arousal and attention distraction on the control of spontaneous postural sway. During each task, postural activity was quantified in terms of center-of-pressure displacement, leg-muscle activation, and ankle and hip rotation. Arousal was monitored via skin conductance, and questionnaires were used to assess state anxiety. Respiratory trunk movements were also recorded, to assess potential arousal-related destabilizing effects, but failed to show a strong association with the postural measures. The nature of the secondary task affected the postural responses in that subjects tended to lean slightly forward, and exhibited higher levels of activation in tibialis anterior, when performing a mental-arithmetic task. Although this task affected both attention and arousal, the leaning effect was limited to subjects who reported higher levels of anxiety related autonomic or somatic symptoms during testing, and the degree of leaning was correlated with the level of physiological arousal. A secondary task that diverted attention but did not affect physiological arousal (listening to a spoken-word recording) failed to elicit changes in postural response. These findings suggest that the changes in leaning were associated with task-related changes in physiological arousal, and highlight the need to account for the potentially confounding influence of arousal when studying attentional effects. Given the potential influence on stability, the results also suggest that leaning should be monitored or controlled during balance assessment, particularly when dealing with individuals who may be anxious or afraid of falling. PMID- 8719512 TI - Reactogenicity and immunogenicity of a three-component acellular pertussis vaccine administered as the primary series to 2, 4 and 6 month old infants in the United States. AB - Five hundred and fifty-seven infants received either an acellular pertussis (DTaP) vaccine containing pertussis toxoid (PT), filamentous hemagglutinin (FHA) and pertactin (PRN) or one of two commercially available whole-cell pertussis (DTP) vaccines at 2, 4 and 6 months. One month after the third immunization, IgG antibody values to pertussis toxoid, filamentous hemagglutinin and PRN were significantly greater following DTaP than either DTP (P < 0.05). When reactions within 48 h after all three doses of vaccine were combined, fever 101 degrees, > or = moderate fussiness, > or = moderate pain, swelling 10 mm, and erythema 10 mm occurred less often after DTaP compared with DTP-Connaught (P < 0.001). The same adverse events were also less after DTaP compared with DTP-Lederle (P < 0.05), except for erythema 10 mm. This three-component DTaP vaccine produced fewer adverse events and greater antibody values to PT, FHA and PRN in comparison with either licensed DTP vaccine when given as the primary series. PMID- 8719513 TI - Recombinant and virion-derived soluble and particulate immunogens for vaccination against tick-borne encephalitis. AB - Using different forms of the envelope glycoprotein E from tick-borne encephalitis virus we investigated the influence of physical and antigenic structure on the efficacy of vaccination. Different protein E-containing preparations were either derived from purified virions or were produced as recombinant proteins in COS cells. These included soluble dimeric forms (virion-derived protein E dimers with and without membrane anchor; recombinant protein E dimers without membrane anchor), micellar aggregates of protein E (rosettes), and recombinant subviral particles (RSPs). The structural differences between these immunogens were verified by sedimentation analysis, immunoblotting and epitope mapping with a panel of monoclonal antibodies. Specific immunogenicities were determined in mice in comparison to formalin-inactivated whole virus. Rosettes and RSPs were excellent immunogens and exhibited similar efficacies as inactivated virus in terms of antibody induction and protection against challenge, whereas all of the soluble forms were much less immunogenic. These data emphasize the importance of the immunogen's antigenic and physical structure for an effective stimulation of the immune system and indicate that RSPs represent an excellent candidate for a recombinant vaccine against tick-borne encephalitis. PMID- 8719514 TI - Mucosal and systemic immunogenicity of a recombinant, non-ADP-ribosylating pertussis toxin: effects of formaldehyde treatment. AB - The effect of formaldehyde treatment on the mucosal and systemic immunogenicity of the genetically detoxified pertussis toxin (PT-9K/129G) was investigated. Groups of BALB/c were immunized intranasally (i.n.) or subcutaneously (s.c.) with untreated, lightly formaldehyde treated (LFT) or heavily formaldehyde treated (HFT) recombinant pertussis toxin (PT) mutant, PT-9K/129G. Intranasal immunization with native PT-9K/ 129G induced significant levels of anti-toxin antibodies in serum and IgA anti-toxin responses in nasal and lung lavages of these mice. Similar local and systemic responses were observed following intransal immunization with LFT toxin. However, i.n. immunization with HFT toxin failed to induce a local IgA response and elicited a much diminished anti-toxin response in the serum. In contrast, the total antibody response following s.c. immunization was not significantly affected. In addition, i.n. immunization with native PT-9K/129G induced low but detectable levels of toxin neutralizing antibodies in the serum. These results show that native PT-9K/129G protein acts as a mucosal immunogen in mice and that this activity is greatly diminished by HFT of the protein. PMID- 8719515 TI - Immunological parameters associated with antigenic competition in a multivalent footrot vaccine. AB - A murine model for antigenic competition with multivalent D. nodosus pili vaccine has been established that parallels the phenomenon observed in sheep where levels of antibody, specific for any particular serogroup of pili, are significantly lower following vaccination in the presence of multiple serogroups of pili than with that serogroup alone. This competition was observed in both high and low responder strains of mice and was not dependent on the multiplicity of the antigens in the multivalent vaccine but could be observed with a large excess of a single heterologous serogroup. Competition was manifest by a reduction in the number of serogroup-specific antibody secreting cells elicited in response to vaccination. The antibody response to a single serogroup of pili reached a plateau at high doses and it was at these doses that antigenic competition was most pronounced, under conditions where both B- and T-cell responses were limiting. The limit in T-cell responsiveness was not imposed at the level of presentation of antigen. Pili-specific T cells were largely cross-reactive for different serogroups, and under conditions of limiting T-cell stimulation within a lymph node the available T cells would have to be shared between B cells specific for each serogroup of pili, which may in turn result in the decrease of serogroup-specific antibody induced following inoculation with the multivalent vaccine. PMID- 8719516 TI - Serum and colostrum antibody responses induced by jet-injection of sheep with DNA encoding a Cryptosporidium parvum antigen. AB - In an effort to generate high titer colostrum for immunotherapy of cryptosporidiosis, a study was conducted to test the efficacy of immunizing sheep with recombinant plasmid DNA (pCMV-CP15/60) encoding epitopes of 15 and 60 kDa surface antigens of Cryptosporidium parvum sporozoites. The plasmid DNA was used to immunize preparturient ewes at three dose levels by jet-injection into either hind limb muscle (IM) or mammary tissue (IMAM). Regardless of route of injection, a dose-dependent anti-CP15/60 immunoglobulin response was observed in sera and colostrum from sheep immunized with pCMV-CP15/60 plasmid DNA. High titer antibody responses were observed in one of three animals per group receiving an IM injection of 100 or 1000 micrograms pCMV-CP15/60. IMAM immunization with 100 or 1000 micrograms pCMV-CP15/60 plasmid DNA elicited higher titer colostrum responses and more consistent serum responses compared to IM injections. A negligible serum and colostrum anti-CP15/60 response was observed in ewes injected IM with 10 micrograms pCMV-CP15/60 or 1000 micrograms control plasmid DNA. Immunoblotting of native C. parvum sporozoite/oocyst protein with hyperimmune serum and colostrum corroborated the increased titers against CP15/60 antigen. Serum and colostrum antibodies from pCMV-CP15/60-immunized sheep were eluted from native CP15 protein and bound a surface antigen of C. parvum sporozoites as indicated by indirect immunofluorescence staining. PMID- 8719517 TI - Comparative study of the immunogenicity and safety of Engerix-B administered at 0, 1, 2 and 12 months and Recombivax HB administered at 0, 1, and 6 months in healthy adults. AB - A randomized trial compared the safety and immunogenicity of Engerix-B (EB) 20 micrograms administered intramuscularly (IM) at 0, 1, 2, and 12 months with Recombivax HB (RHB) 10 micrograms administered IM at 0, 1, and 6 months in healthy adults. At months 3 and 6, significantly more subjects who received EB were seroprotected compared to those who received RHB (84 vs 67%, p = 0.0027; 95 vs 76%, p < 0.001, respectively). SP rates were similar between the vaccination groups approximately 1 year after administration of the initial dose (91 and 83%, respectively; p = 0.1). The vaccines were well tolerated with injection site pain being the most commonly reported adverse event. PMID- 8719518 TI - Priming for virus-specific CD8+ but not CD4+ cytotoxic T lymphocytes with synthetic lipopeptide is influenced by acylation units and liposome encapsulation. AB - Synthetic peptides of the herpes simplex virus glycoprotein B synthesized either as a free form or derivatized with one (PAM1) or three palmitic acids (PAM3Cys) were used to assess the in vivo priming efficacy of high affinity virus-specific CTL induction. The peptide and its derivatives were delivered in vivo with or without liposome encapsulation. Neither the free peptide nor the PAM1 derivative primed for high affinity virus specific CD8+ CTL induction, whether delivered via liposomes or not. On the other hand, the PAM3Cys derivative was able to prime for low levels of high affinity virus specific CD8+ CTL induction in the absence of liposome encapsulation. However, the efficiency of virus-specific CD8+ CTL induction with PAM3Cys derivative was enhanced following encapsulation in the liposomes. In contrast, all forms of the peptides induced both CD4+ T cell proliferative response as well as high affinity virus-specific CD4+ CTL. In addition, the efficiency of the PAM3Cys derivative to prime for CD4+ or CD8+ CTL was found to be influenced by the liposome encapsulation. When delivered via liposomes, the PAM3Cys derivative effectively primed for CD8+ CTL. However, liposomal delivery was not necessary for efficient priming for CD4+ CTL induction. Thus, both the acylation units as well as liposomal delivery appear to influence the in vivo priming of CD4+ and CD8+ T cell responses with synthetic peptides. PMID- 8719519 TI - Protection of ruminants by Pasteurella haemolytica A1 capsular polysaccharide vaccines containing muramyl dipeptide analogs. AB - The capsular polysaccharide (CP) of Pasteurella haemolytica serotype A1 is a poor immunogen for the prevention of pneumonic pasteurellosis of ruminants. To improve CP immunogenicity, vaccines were prepared with 1.0 mg CP dose-1 with and without the synthetic adjuvant, muramyl dipeptide (MDP; range 0.2-1.0 mg) or a lipophilic derivative, MDP-sn-glyceryl-dipalmitoyl (MDP-GDP; range 0.1-1.0 mg). The optimum effective concentration of adjuvant was first determined in lambs and calves and then the efficacy of CP +0.5 mg MDP and CP +1.0 mg MDP-GDP was compared with that of two commercial vaccines in calves. After immunization with CP, antibody titers in lambs and calves were typical of that seen with polysaccharide immunogens and characterized by an early IgM response followed by later IgG1 and IgG2 responses. CP + MDP or MDP-GDP vaccines induced significantly higher IgM, IgG1, and IgG2 titers. After transtracheal challenge of immunity with P. haemolytica serotype A1, extensive pulmonary consolidation containing P. haemolytica (10(6)-10(8) c.f.u. g-1) was seen in all lambs and calves vaccinated with CP alone and was not significantly different (P < 0.05) from the consolidation and concentrations of organisms in nonvaccinated challenge controls. In lambs, vaccines containing 1.0 mg CP +0.05 mg MDP or MDP-GDP significantly reduced pulmonary consolidation and concentrations of P. haemolytica in lung lesions. In calves, vaccines containing 0.2 mg MDP, 0.5 mg MDP, or 1.0 mg MDP-GDP also significantly reduced pulmonary consolidation and concentrations of P. haemolytica in lung lesions. Vaccines containing CP +0.5 mg MDP and CP +1.0 mg MDP-GDP induced high titer bactericidal antibodies by 7 days and were more efficacious than two commercial vaccines. Potentiation of CP with MDP or MDP-GDP has great promise in furthering the potential of CP as a vaccine immunogen for the prevention of pneumonic pasteurellosis. PMID- 8719520 TI - A follow-up study of combined vaccination with plasma-derived and recombinant hepatitis B vaccines in infants. AB - This study was aimed to evaluate the efficacy and immunogenicity of combined hepatitis B vaccination with plasma-derived vaccine (PDV) and recombinant vaccine (RV). A total of 329 infants was recruited, including 224 high-risk infants born to hepatitis B e antigen-positive mothers and 105 low-risk infants born to hepatitis B e antigen-negative mothers. The high-risk infants received four doses of hepatitis B vaccine at 0, 1, 2 and 12 months of age with five different schedules. Group A1 and A2 infants were vaccinated with PDV as the first dose and RV (SB vaccine for group A1, MSD vaccine for group A2) as the remaining three doses. Group B1 and group B2 infants were vaccinated with PDV as the first two doses and RV (SB vaccine for group B1, MSD vaccine for group B2) as the remaining two doses. Group C infants received four doses of PDV. Low-risk infants were vaccinated with PDV at birth, and RV at 1 and 6 months of age (group D1, using SB vaccine; group D2, using MSD vaccine). At completion of vaccination schedules, 20 of 224 high-risk infants (9%) were positive for hepatitis B surface antigen. The overall protective efficacy was 90%. Hepatitis B surface antibody (anti-HBs) positive rate ranged between 94 and 100% among the remaining infants. The protective efficacy and immunogenicity were similar among groups except that the mean level of anti-HBs in group C, D1 and D2 infants tended to be lower than that of the other four groups. To ensure an optimal immune response, four doses of vaccine are recommended in high-risk infants when two types of vaccine are to be used in combination. PMID- 8719521 TI - Isolation and quantification of Quillaja saponaria Molina saponins and lipids in iscom-matrix and iscoms. AB - In the iscom, multiple copies of antigen are attached by hydrophobic interaction to a matrix which is built up by Quillaja triterpenoid saponins and lipids. Thus, the iscom presents antigen in multimeric form in a small particle with a built-in adjuvant resulting in a highly immunogenic antigen formulation. We have designed a chloroform-methanol-water extraction procedure to isolate the triterpenoid saponins and lipids incorporated into iscom-matrix and iscoms. The triterpenoids in the triterpenoid phase were quantitated using orcinol sulfuric acid detecting their carbohydrate chains and by HPLC. The cholesterol and phosphatidylcholine in the lipid phase were quantitated by HPLC and a commercial colorimetric method for the cholesterol. The quantitative methods showed an almost total separation and recovery of triterpenoids and lipids in their respective phases, while protein was detected in all phases after extraction. The protein content was determined by the method of Lowry and by amino acid analysis. Amino acid analysis was shown to be the reliable method of the two to quantitate proteins in iscoms. In conclusion, simple, reproducible and efficient procedures have been designed to isolate and quantitate the triterpenoids and lipids added for preparation of iscom-matrix and iscoms. The procedures described should also be useful to adequately define constituents in prospective vaccines. PMID- 8719522 TI - Construction and immunogenicity of Salmonella typhimurium vaccine vectors that express HIV-1 gp120. AB - Since the human immunodeficiency virus (HIV-1) is transmitted either parenterally or sexually, both mucosal and systemic immune responses may be required to provide protective immunity. Attenuated Salmonella vectors expressing heterologous antigen can stimulate responses in both compartments. To evaluate the utility of Salmonella vectors as an HIV-1 vector vaccine, a gene expression cassette encoding recombinant HIV-1 gp120 (rgp120) was integrated into the hisOGD locus of Salmonella typhimurium aroA strain, SL3261 (SL3261::120). To test if increased antigen expression potentiates immunogenicity, strains were constructed that express rgp120 from a multicopy asd-stabilized plasmid (SL7207 pYA:120). Immunoblot analysis demonstrated that SL7207 pYA:120 expressed approximately 50 fold more rgp120 than SL3261::120. Oral immunization of BALB/c mice with these strains did not stimulate an env-specific CTL response or a significant rise in antigp120 antibody titer as compared to controls. However, splenic T cells from SL7207 pYA::120 immunized mice proliferated upon restimulation with gp120 in vitro while splenocytes from SL3261::120 immunized mice did not, gp120 restimulated splenic T cells from SL7207 pYA:120 immune mice also produced IFN gamma but no IL-5. Two conclusions can be drawn from these results. First, high level expression of rgp120 in Salmonella vectors is necessary to stimulate a gp120-specific immune response in mice. Second, Salmonella::rgp120 stimulates a gp120-specific Th1 response in mice. This is the first report to describe the construction of a Salmonella::rgp120 vector vaccine that is immunogenic in mice. PMID- 8719523 TI - Immunomodulatory effects of HSV2 glycoprotein D in HSV1 infected mice: implications for immunotherapy of recurrent HSV infection. AB - Immunological analyses in this laboratory and others have suggested that a nonrecurrent HSV seropositive immune status is more closely correlated with a type 1 T helper cell (Th1) response characterized by elevated levels of interferon-gamma and IL2 rather than high titers of virus-specific antibodies. Effective intervention with an immunotherapeutic vaccine may require modulation of the regulatory network of T helper cells such that there is selective restimulation and expansion of the Th1 response. We have established a murine model for assessing the immunomodulatory capacity of an HSV glycoprotein subunit vaccine in animals with pre-existing herpes immunity. Animals were infected with varying doses of HSV1 and then administered glycoprotein D (gD) vaccine adjuvanted with aluminum phosphate at 3-week intervals. Observed changes in serological and cellular responses indicated that administration of subunit vaccine adjuvanted with aluminum phosphate could shift a dominant Th1 response, induced by sensitization with live HSV, towards a Th2 profile of activity. These data suggest that use of aluminum based adjuvants will not selectively stimulate Th1-associated responses and alternative adjuvants may be required for effective use of subunit vaccine in an immunotherapeutic indication in humans. PMID- 8719524 TI - Antigenic analysis of Japanese encephalitis viruses in Asia by using monoclonal antibodies. AB - Based on our previous study using monoclonal antibodies against three Japanese encephalitis (JE) virus strains, Nakayama-RFVL, Beijing 1 and Kamiyama, 25 JE virus strains isolated mainly in Japan and some from Southeast Asia between 1935 and 1979 fell into five antigenic groups, Nakayama, Beijing 1, Kamiyama, Muar and 691004. In the present study, monoclonal antibodies against Muar and 691004 strains were produced to analyze immunological characteristics of JE virus in detail. We obtained the Muar group-specific antibody and three anti-691004 monoclonal antibodies which showed different reactivities. Furthermore, antigenic comparison of the 22 JE virus strains isolated in Asia between 1963 and 1984 was performed by using the 15 monoclonal antibodies characterized by the different reactivities against the above five antigenic groups. Of these 22 strains, the 17 strains reacted with the Kamiyama group-specific monoclonal antibody, but anti Nakayama, anti-Beijing 1 and anti-Muar group-specific monoclonal antibodies showed no reactivities with any of the strains. This suggested that the currently prevalent JE virus strains in Asia belonged to the Kamiyama group. The other five strains, ThCMP 1982, KE083, KE093, 733913 and Ling, did not react with the above four group-specific monoclonal antibodies, but all except the KE093 strain showed a similar pattern to the Kamiyama strain on the basis of the reactivities against the other 11 antibodies. The KE093 strain isolated from Thailand in 1983 showed immunologically outstanding difference from other strains. PMID- 8719525 TI - Myxoma virus as a vaccine vector for rabbits: antibody levels to influenza virus haemagglutinin presented by a recombinant myxoma virus. AB - To determine whether myxoma virus (MV) could be used as a vector for antigen presentation to European rabbits (Oryctolagus cuniculus), despite its immunosuppressive properties, a recombinant MV expressing the influenza virus haemagglutinin (HA) was constructed. Domestic rabbits, inoculated with the recombinant virus, developed high plasma antibody titres to the HA and IgG to HA was also detected in vaginal secretions. Antibody titres to HA obtained with recombinant MV were similar to titres obtained in rabbits inoculated with vaccinia virus expressing the HA. These results indicate that MV is an efficient vector for antigen presentation in rabbits. PMID- 8719526 TI - Urgent need for effective vaccine against Salmonella paratyphi A, B and C. PMID- 8719527 TI - Ethical guidelines for pain research in humans. Committee on Ethical Issues of the International Association for the Study of Pain. PMID- 8719528 TI - Efficacy of epidural steroid injections for low-back pain and sciatica: a systematic review of randomized clinical trials. AB - The purpose of the study was to assess the efficacy of epidural steroid injections for low-back pain. Data was obtained using computer-aided search of published randomized clinical trials and assessment of the methods of the studies. Twelve randomized clinical trials evaluating epidural steroid injections were identified. Data was extracted based on scores for quality of the methods, using 4 categories (study population, interventions, effect measurement, and data presentation and analysis) and the conclusion of the author(s) with regard to the efficacy of epidural steroid injections. Method scores of the trials ranged from 17 to 72 points (maximum 100 points). Eight trials showed method scores of 50 points or more. Of the 4 best studies (> 60 points), 2 reported positive outcomes and 2 reported negative results. Overall, 6 studies indicated that the epidural steroid injection was more effective than the reference treatment and 6 reported it to be no better or worse than the reference treatment. There appeared to be no relationship between the methodological quality of the trials and reported outcomes. In conclusion, there are flaws in the design of most studies. The best studies showed inconsistent results of epidural steroid injections. The efficacy of epidural steroid injections has not been established. The benefits of epidural steroid injections, if any, seem to be of short duration only. Future research efforts are warranted, but more attention should be paid to the methods of the trials. PMID- 8719529 TI - Immune activation: the role of pro-inflammatory cytokines in inflammation, illness responses and pathological pain states. AB - It has recently become accepted that the activated immune system communicates to brain via release of pro-inflammatory cytokines. This review examines the possibility that pro-inflammatory cytokines (interleukins and/or tumor necrosis factor) mediate a variety of commonly studied hyperalgesic states. We will first briefly review basic immune responses and inflammation. We will then develop the concept of illness responses and provide evidence for their existence and for the dramatic changes in neural functioning that they cause. Lastly, we will examine the potential roles that both pro-inflammatory cytokines and the neural circuits that they activate may play in the hyperalgesic states produced by irritants, inflammatory agents, and nerve damage. The possibility is raised that apparently diverse hyperalgesic states may converge in the central nervous system and activate similar or identical neural circuitry. PMID- 8719530 TI - Intrathecal prostaglandin E1 produces a long-lasting allodynic state. AB - The existence of prostaglandin (PG) receptors in the spinal cord has been demonstrated, but their role in sensory processing is not yet well defined. PGE1 is widely used clinically as a vasodilator. The present study was designed to investigate the effects of intrathecally administered PGE1 on the transmission of different types to sensory information, including that associated with noxious somatic, noxious visceral, and non-noxious somatic stimulation. The tail-flick (TF) test was employed to measure responses to noxious somatic stimuli, and the colorectal distension test was used to examine responses to noxious visceral stimuli. Withdrawal response to mechanical pressure produced by Semmes-Weinstein mono-filaments (SWMs) was measured as an assessment of sensitivity to non-noxious mechanical somatic stimulation. TF latencies and colorectal distension thresholds decreased for a short time (10-20 min) following the intrathecal (i.t.) administration of both 100 ng or 500 ng of PGE1. In sharp contrast to these short duration effects, there was a long-lasting increase in agitation scores (allodynia) produced by 3 different intensities of SWMs (0.217, 0.745 and 2.35 g) after administration of PGE1. The changes in agitation scores to SWMs were dependent on the dose of PGE1 and the intensity of stimulation. This increase of agitation score was seen when PGE1 was administered through the i.t. catheter or by direct i.t. puncture and the increase lasted for at least 2 days after drug administration. Intrathecal administration of saline, however, did not produce any changes in TF latencies, colorectal distension thresholds, or agitation scores produced by SWMs. No significant histological difference was seen between spinal cords exposed to 500 ng PGE1 and saline 48 h after drug administration. These results demonstrate that PGE1 may trigger a hypersensitive (allodynic and/or hyperalgesic) state in sensory processing pathways at the spinal level. They also indicate that long-lasting changes in processing of non-noxious, but not noxious, information produced by PGE1 continues after the disappearance of the direct action of PGE1. PMID- 8719531 TI - Attenuation of pain-related hyperventilation in adjuvant arthritic rats with adrenal medullary transplants in the spinal subarachnoid space. AB - The adjuvant arthritic rat model has been utilized for the study of chronic pain, as polyarthritic rats present a variety of symptoms similar to those seen in human chronic pain conditions. In particular, hyperventilatory responses are notable in both and may more accurately reflect basal ongoing pain than do evoked noxious stimuli. To assess whether adrenal medullary transplants in the spinal subarachnoid space can alleviate basal arthritic pain, respiratory parameters were determined using whole body plesthmography in polyarthritic rats. Arthritis was induced by inoculation with an intradermal injection of complete Freund's adjuvant. Steady-state ventilation was monitored at weekly intervals in arthritic animals with adrenal medullary or control striated muscle transplants. Results revealed that adjuvant arthritis produced significant hyperventilation in animals with control transplants, as indicated by increased tidal volumes and minute ventilation, which paralleled the progression of the inflammatory process. In contrast, this hyperventilation was eliminated by adrenal medullary transplants. A role for catecholamines and opioid peptides released from the transplants was suggested by the reversal of these effects with phentolamine and naloxone. In addition, the retardation in weight gain normally observed in polyarthritic animals was markedly attenuated by adrenal medullary, but not control transplants. These findings indicate that adrenal medullary transplants in the spinal subarachnoid space can alleviate basal chronic pain as assessed in adjuvant arthritis. PMID- 8719532 TI - Neonatal testosterone exposure influences neurochemistry of non-opioid swim stress-induced analgesia in adult mice. AB - The effects of neonatal hormone manipulations on swim stress-induced analgesia (SSIA) magnitude and neurochemical quality were examined in Swiss-Webster mice of both sexes. Previous research has indicated that non-opioid SSIA mechanisms in adult Swiss-Webster mice are sexually dimorphic. Male mice exhibit non-opioid SSIA following a 3-min swim in cold (15 degrees C) water that is antagonized by the non-competitive NMDA antagonist MK-801 (dizocilpine; 0.075 mg/kg), whereas female mice do not display NMDA-mediated analgesia in the presence of estrogen. Since male and female mice show equipotent magnitudes of SSIA, it was concluded that female mice display a neurochemically distinct, estrogen-dependent SSIA mechanism specific to their gender. In the present study, female mice exposed to testosterone during the neonatal period display NMDA-mediated analgesia even in the presence of estrogen in adulthood. Thus, expression of the female-specific, estrogen-dependent SSIA mechanism previously described may be dependent on the absence of testosterone during early ontogeny. PMID- 8719533 TI - Sex differences in the expression and antagonism of swim stress-induced analgesia in deer mice vary with the breeding season. AB - Swim stress-induced analgesia (SSIA) was examined in photoperiodically induced 'breeding' (reproductive) and 'non-breeding' (non-reproductive) adult male and female deer mice, Peromyscus maniculatus. Nociceptive responses (50 degrees C, hot-plate) of breeding and non-breeding deer mice were determined after either a 1- or 3-min swim in 20 degrees C water. The 1-min swim induced an immediate and relatively short-lasting naloxone (1.0 mg/kg) insensitive 'non-opioid' -mediated SSIA that was antagonized by the N-methyl-D-aspartate (NMDA) antagonist, MK-801 (0.10 mg/kg) in all of the groups of mice except the breeding (reproductive) females. Breeding females displayed a non-opioid analgesia that was insensitive to MK-801. The 3-min swim induced a relatively more prolonged mixed opioid and 'non-opioid' SSIA of which the initial portion was sensitive to antagonism by MK 801 in all groups of the mice except the breeding females, while the latter portion (15 min after swim) was induced by naloxone in all of the groups of mice. Overall, the breeding males displayed greater levels of SSIA than the breeding females, with no consistent sex differences in the non-breeding mice. Within sexes, the breeding males displayed greater levels of opioid and non-opioid SSIA than the non-breeding males, while the non-breeding females displayed greater levels of SSIA than the breeding females. These results show that both sex and reproductive status affect the expression and neurochemical mediation of non opioid SSIA. These findings also suggest that reproductive females may have an unique or novel hormonally (estrogen) dependent mechanism associated with the expression of SSIA. PMID- 8719534 TI - Increased pressure pain sensibility in fibromyalgia patients is located deep to the skin but not restricted to muscle tissue. AB - This study was aimed at comparing pressure pain sensibility in different tissues in fibromyalgia patients. Pressure pain thresholds (PPTs) were assessed in 16 fibromyalgia (FM) patients bilaterally at the bony part of epicondylus lateralis humeri, at the belly of m. extensor carpi ulnaris and at m. brachioradialis where the radial nerve branches pass underneath. Following a double-blind design, either a local anesthetic cream (EMLA) or a control cream was applied to the skin and PPTs were reassessed. The site with underlying nerve had a lower PPT than the bony site (P < 0.001) and the 'pure' muscle site (P < 0.001), respectively. These relations remained unaltered by skin hypoesthesia. The PPTs over the bony and the 'pure' muscle sites did not differ. Application of EMLA, compared to control cream, did not change PPTs over any area examined. The results demonstrated that pressure-induced pain sensibility in FM patients is not most pronounced in muscle tissue and does not depend on increased skin sensibility. PMID- 8719535 TI - Sensitivity of patients with painful temporomandibular disorders to experimentally evoked pain. AB - Temporomandibular disorders (TMD) represent a group of chronic painful conditions involving the muscles of mastication and the temporomandibular joint. We determined whether patients with painful TMD are more sensitive to noxious stimuli than age-matched control subjects. Fifty-two TMD patients (16 with muscle pain and 36 with combined muscle and joint pain) and 23 age-matched and gender matched volunteers participated. Forearm thermal pain threshold and tolerance values were determined. A submaximal effort tourniquet procedure was used to evoke ischemic muscle pain. Relative to control subjects, TMD patients had significantly lower thermal pain threshold, ischemic pain threshold, and ischemic pain tolerance values; and thermal pain tolerance values also tended to be lower. Pain sensitivity did not differ between the two groups of TMD patients. Furthermore, the submaximal effort tourniquet procedure, which is capable of altering acute orofacial pain (Sigurdsson and Maixner, 1994) did not produce a consistent reduction in orofacial pain associated with TMD. We concluded that TMD patients are more sensitive to noxious stimuli than pain-free controls. These findings provide additional evidence that TMD is a psychophysiological disorder of the central nervous system which modulates emotional, physiological and neuroendocrine responses to emotional and physical stressors. PMID- 8719536 TI - Chronic pain patient-spouse behavioral interactions predict patient disability. AB - Based on behavioral theory, it has been hypothesized that spouse solicitous responses to the pain behaviors of chronic pain patients may contribute to the maintenance of pain behaviors and disability. Self-report data support this hypothesis, but direct observational measures have not been used to study this association. In this study, 50 chronic pain patients and their spouses were videotaped while engaging in common household activities. and patient pain behaviors and spouse solicitous behaviors were coded from the tapes. Spouse solicitous responses to non-verbal pain behaviors were significant predictors of physical disability in the more depressed patients, and were significant predictors of rate of non-verbal pain behavior in patients who reported greater pain. Spouse solicitous responses did not predict psychosocial dysfunction or total self-reported pain behaviors. The result support behavioral theory and indicate the need for further study of the association between spouse solicitousness and patient pain behaviors/disability. PMID- 8719537 TI - A clinical procedure for assessment of severity of knee pain. AB - We describe a clinical procedure for assessing knee pain: 10 standardised movements of the knee are made (r active and 6 passive), and the subject's behavioural response to each is scored by the assessor on a 0-3 scale. The total score is the Pain Index of the Knee (PIK). We report investigations of repeatability, inter-assessor agreement, and validity of components of the PIK compared with VAS ratings by the subject. We conclude from these results and from our experience of the PIK in practice that it is useful both clinically and for research purposes. It is a simple and efficient procedure for pain measurement in terms of behavioural response, with good reliability and validity, at least at low and medium levels of pain. PMID- 8719538 TI - Aspirin and acetaminophen reduced both Fos expression in rat lumbar spinal cord and inflammatory signs produced by carrageenin inflammation. AB - This study, performed in freely moving rats, evaluates the effects of the two most prescribed analgesics, aspirin and acetaminophen, on carrageenin inflammation and the associated c-Fos expression in the rat lumbar spinal cord. Maximal dorsal horn c-Fos expression is observed 3 h after carrageenin (6 mg/150 microliters of saline), with Fos-like (Fos-LI) neurones being predominantly located in laminae I-II and V-VI (41 +/- 3% and 39 +/- 5% of the total number of Fos-LI neurones per section for the control group, respectively) of the dorsal horn. Pretreatment with aspirin (75 or 150 mg/kg, i.v.) reduced the number of Fos LI neurones induced by carrageenin-inflammation (28 +/- 2% and 45 +/- 1% reduction, respectively; P < 0.001 for both). Acetaminophen (75 or 105 mg/kg, i.v.) reduced the number of Fos-LI neurones (19 +/- 1% and 43 +/- 1% reduction, respectively; P < 0.001 for both). When considering the lower dose (75 mg/kg), the effects of aspirin were significantly more marked than those of acetaminophen (P < 0.001). There was a tendency for both aspirin and acetaminophen to have a more pronounced effect on the number of Fos-LI neurones located in deeper laminae, these differential effects being significant for 75 mg/kg of aspirin (P < 0.01) and 150 mg/kg of acetaminophen (P < 0.01). Both the two doses of aspirin and acetaminophen greatly reduced the inflammatory signs associated with the intraplantar injection of carrageenin. Furthermore there was a positive correlation between the effects of aspirin and acetaminophen on the number of Fos LI neurones and the inflammatory signs which developed after carrageenin. Our results suggest that the effects of both drugs are mainly due to peripheral site of action without rejecting an additional central site of action of systemic non steroidal anti-inflammatory drugs (NSAIDs) and acetaminophen. In addition, our results suggest that the approach we used could be a useful tool to evaluate systematically and quantitatively the effects of NSAIDs. Finally, the effects obtained with the low dose of acetaminophen question the classical view of textbooks claiming that such a compound had no anti-inflammatory effect and are in agreement with previous observations in humans. PMID- 8719539 TI - Activation of a NO-cyclic GMP system by NO donors potentiates beta-endorphin induced antinociception in the mouse. AB - Nitric oxide (NO) donors such as sodium nitroprusside (SNP, 0.01-1 micrograms) or 3-morpholino-sydnonimine (SIN-1, 0.1-10 micrograms) administered intracerebroventricularly (i.c.v) produced a dose-dependent potentiation of beta endorphin-induced antinociception assessed by the tail-flick test in ICR mice. The same i.c.v. treatment with SNP or SIN-1 did not affect the antinociception induced by mu-, delta-, or kappa-opioid receptor agonists. The goal of the present study was to determine if the potentiation of the beta-endorphin-induced antinociception by NO donors is mediated by the activation of NO-cGMP system. Co administration of hemoglobin (30-120 micrograms) or methylene blue (1.25-5 micrograms), but not N omega-nitro-L-arginine (1-5 micrograms) given i.c.v. dose dependently attenuated the potentiating effects of SNP or SIN-1 on beta-endorphin induced antinociception. However, the same i.c.v. treatments of mice with hemoglobin, methylene blue or N omega-nitro-L-arginine did not directly affect the i.c.v. administered beta-endorphin-induced antinociception. On the other hand, the treatment of mice with a combination of NO donor (SNP, 0.1 micrograms or SIN-1, 1 microgram) and zaprinast (a cGMP phosphodiesterase inhibitor, 1 microgram) further potentiated beta-endorphin-induced antinociception. These results indicate that the potentiating effect of SNP or SIN-1 on beta-endorphin induced antinociception is mediated by the increased production of NO-cyclic GMP in the brain. However, the NO-cGMP system is not directly involved in the beta endorphin-induced antinociception. PMID- 8719540 TI - Relationship between hypothalamic-pituitary-adrenal activity and blockade of tolerance to morphine analgesia by pain: a strain comparison. AB - We previously reported that morphine fails to produce analgesic tolerance when administered in the presence of formalin-induced pain. The hypothalamic-pituitary adrenal (HPA) axis is known to respond to stressful stimuli, including pain. To examine whether the blockade of tolerance by pain is related to HPA activity, we assessed the development of tolerance to morphine analgesia in an inbred strain of rats that lack typical stress-induces HPA responses (Lewis strain). Lewis rats lack typical stress-induced activation of corticotropin-releasing hormone, adrenocorticotropin hormone and glucocorticoids. Female Lewis rats were injected with morphine (20 mg/kg, i.p.) or saline for 4 consecutive days in the presence or absence of pain induced by injection of formalin into the hind-paw. The analgesic effect of morphine (5, 10 or 20 mg/kg, i.p.) was then measured in the tail-flick test 24 h after tolerance induction. Inbred female Fischer rats, which show significant stress-induced HPA activity, were used for comparison. Analgesic tolerance was produced in both strains when morphine was delivered in the absence of pain. However, the presence of pain during morphine administration prevented the development of analgesic tolerance in Fischer, but not in Lewis, rats. The differential of pain on the development of tolerance to morphine analgesia are suggested to be related to genetically determined differences in stress-induced HPA activity. PMID- 8719543 TI - Phantom or central neuropathic pain? PMID- 8719542 TI - Hypnotic analgesia and reflex activity. PMID- 8719544 TI - Comments on Schwarzer et al., PAIN, 58 (1994) 195-200. PMID- 8719545 TI - The significance of glomerular hypertrophy in focal segmental glomerulosclerosis. AB - It is not clear whether glomerular hypertrophy is linked to the pathogenesis of human focal segmental glomerulosclerosis (FSGS). To probe the significance of glomerular hypertrophy in relation to development of FSGS, we studied 16 adults with primary FSGS by morphometry, and the findings were compared to age- and sex matched subjects with minimal lesion. Mean glomerular volume (MGV), mesangial volume density, mesangial volume per glomerulus, and cortical interstitial volume density [Vv(int/cortex)] were significantly increased in the FSGS patients when compared to the minimal lesion patients. The increase in mesangial volume in FSGS was mainly due to expansion of mesangial matrix. In FSGS, MGV was related directly to % of glomeruli with glomerulosclerosis (r = 0.47, p < 0.05), to mesangial volume per glomerulus (r = 0.57, p < 0.01), and to Vv(int/cortex) (r = 0.47, p < 0.05). The percentage of glomerulosclerosis correlated directly with Vv(int/cortex) (r = 0.83, p < 0.0005), and with mesangial volume per glomerulus (r = 0.47; p < 0.05) in FSGS. Also, there was a direct relationship between Vv(int/cortex) and mesangial volume per glomerulus (r = 0.49; p < 0.05) in FSGS. Glomerular hypertrophy observed in our patients with primary FSGS was intercorrelated with the extent of glomerulosclerosis, mesangial expansion and interstitial fibrosis. Glomerular hypertrophy seems to be one of the morphological facets present in FSGS. Glomerular hypertrophy tends to coexist with FSGS rather than precede its development. Thus, in biopsies diagnosed with minimal lesion the presence of glomerular hypertrophy appears to be an indication that the coexistent FSGS lesions are undetected as a result of sampling problems. PMID- 8719547 TI - Clinicopathological study of IgA nephropathy in patients with congenitally reduced nephron mass. AB - In experimental animal models, a reduction in the number of functioning nephrons is considered to play a role in the progression of glomerular injury. In human renal diseases, however, whether a superimposed reduction in the number of nephrons causes the exacerbation of preexistent glomerulopathy has not been elucidated. We herein report the results of a clinicopathological study of five patients with IgA nephropathy (IgAN) which occurred in a reduced nephron mass status (four cases of congenital solitary kidney and one case of bilateral hypoplastic kidneys). Four of the five patients had chronic renal failure (CRF) and exhibited a relatively rapid course to CRF as primary IgAN. Renal biopsy revealed that all four of the patients with CRF had glomerular hypertrophy and focal segmental glomerular sclerosis. In addition, two of them had a focal active lesion. In one patient with bilateral hypoplastic kidneys renal biopsies were performed twice in eight years. During this period her creatinine clearance deteriorated from 60.0 ml/min to 20.7 ml/min. Her first renal biopsy showed mild mesangial proliferation without sclerotic lesions, glomerular hypertrophy and mesangial IgA deposition, while all of them were prominent in the second renal biopsy. These observations suggest that IgAN superimposed on a nephron loss status may be frequently associated with a progressive course of disease, and careful follow-up and early treatment should be considered in such a condition. PMID- 8719546 TI - Glomerular metalloprotease activity modulates the development of focal segmental glomerulosclerosis. AB - Focal segmental glomerulosclerosis (FSGS) is a common outcome of a variety of renal diseases. Among laboratory animals both puromycin aminonucleoside (PAN) and hypertension produce a similar histological pattern. Since mesangial expansion is a precursor of FGS and decreased degradation of matrix can cause expansion of mesangium we studied the glomerular matrix metalloprotease activities in the development of FSGS. Dahl salt sensitive (SS) and salt resistant (SR) rats were fed 8% salt diet for six weeks. Kidney biopsy showed features of FSGS in SS rats. Glomeruli were isolated and metalloprotease activity was measured. Sprague-Dawley rats were administered subcutaneously either saline (S) or saline containing PAN (1.67 mg/100 g B. W.) daily for 7 days. Kidney biopsy was done at day 7 and the isolation of glomeruli was performed at day 10 and at 6 weeks. Renal histology showed features of FSGS in PAN rats at 6 weeks. Glomerular metalloprotease activity was decreased in SS (SR, 148 +/- 12 vs. SS, 73 +/- 9 cpm/microgram protein, p < 0.01) as well as in PAN rats (S, 31 +/- 5 vs. PAN, 12 +/- 1 cpm/microgram protein, p < 0.01). These results suggest that decreased glomerular metalloprotease activity may play a role in the development of FSGS in SS and PAN rats. PMID- 8719548 TI - Immunosuppressive treatment of the glomerulonephritis of systemic lupus. AB - To evaluate the results, the long-term prognosis and the rates of complication of an immunosuppressive regimen with corticosteroids and cyclophosphamide in the treatment of the nephritis of systemic lupus erythematosus, 21 patients with lupus glomerulonephritis were studied. Renal biopsies were performed in 17/21 of them and indicated diffuse proliferative (6 patients), diffuse mesangial (4) and membranous (7) glomerulonephritis. Treatment was structured in 4 phases: 1) induction with methylprednisolone 250 mg i.v. for 7-14 days, and cyclophosphamide 100-200 mg p.o., q.d., or 20 mg/kg i.v. every 28 days; 2) maintenance with prednisone p.o., 2 mg/kg q.o.d. for 45 days, and cyclophosphamide as before; 3) tapering, with reduction of prednisone by 15% each month for 4 months; 4) indefinite maintenance with prednisone slowly tapered to the least effective q.o.d. dose and cyclophosphamide discontinued after six months of treatment. This cycle was repeated in the event of a relapse. After a first immunosuppressive cycle, 20/21 patients achieved remission of glomerulonephritis. Plasma creatinine fell from 97 +/- 6 to 80 +/- 3 microMol/l (p < 0.01). Proteinuria fell from 2.1 +/- 0.4 to 0.2 +/- 0.4 g/d (p < 0.0001) and the nephrotic syndrome, present in 8 patients, disappeared. After an average of 20 +/- 7 months, 8 patients relapsed: all remitted again after a repeat cycle, but 1 later progressed to end-stage renal failure during pregnancy. After an average of 56 months 4 out of these 8 patients relapsed again: 1 progressed to end-stage renal disease following an abortion and 3 remitted completely after a third cycle. Thus, 18 out of 21 patients are presently in remission with an average dose of prednisone of 13.7 mg/day after an average follow-up of 52 +/- 38 months (range 2 to 156). Three patients are presently off treatment. In 16 patients with extended follow-up of 2 to 13 years, anti-nuclear antibodies, anti-DNA antibodies, albuminuria and cylindruria fell below post-cycle levels (p < 0.001 for all). We conclude that intensive immunosuppression with steroids and cyclophosphamide can achieve excellent long-term results in the treatment of systemic lupus with glomerulonephritis. PMID- 8719549 TI - P1 blood group phenotype, secretor status in patients with urinary tract infections. AB - We have examined the distribution of P antigen, Lewis blood group phenotypes and secretor status of 65 patients with E. coli UTI (20 asymptomatic bacteriuria, 20 cystitis and normal radiology, 25 reflux nephropathy) and 45 controls who have never experienced a UTI episode. The distribution of Lewis blood group antigens was similar in all UTI groups and in the controls. The incidence of non-secretors in the reflux nephropathy group was similar to that in controls (24% versus 31%). The P1 phenotype was present in 100% of patients with asymptomatic bacteriuria, 80% with cystitis and controls and only 44% with reflux nephropathy. The combined P1/non-secretor phenotype was observed in 45% of patients with asymptomatic bacteriuria, 30% with cystitis, 12% with reflux nephropathy and in 22% of control healthy individuals. P2/secretor phenotype was demonstrated in 44% of patients with reflux nephropathy and in only 11% of controls. Our data suggest that having P2 blood group protects against asymptomatic colonization of the urinary tract, but is associated with the type of infection responsible for scarring in reflux nephropathy. It also appears that being a non-secretor does not predispose to renal scarring and that combined P2/secretor phenotype may be linked with susceptibility to reflux nephropathy. PMID- 8719550 TI - Phosphaturia, glycosuria and aminoaciduria associated with idiopathic acquired sideroblastic anemia. AB - Idiopathic acquired sideroblastic anemia is not a common disease. We studied the renal tubular function in such patients. Patients have lower calcium than controls (8.46 mg/dl +/- 0.59 vs 9.16 +/- 0.53). We have highlighted the multiple renal anomalies observed in patients with this type of hemopathy. The serum phosphate levels are lower in patients than in controls (2.73 +/- 0.36 vs 3.3 +/- 0.55 mg/dl, p = 0.0048). We found higher glycosuria (21:43 +/- 42.58 vs 0.0 +/- 0.0 mg/dl), total aminoaciduria (6280 +/- 3943 vs 4138 +/- 2269 microMol/g creatinine, p = 0.19) and lower maximum capacity for phosphate reabsorption by the renal tube (TmPO4) (2.11 +/- 0.38 vs 2.9 +/- 0.73 mg/100 ml GFR, p = 0.0027) in the patients. The association between idiopathic acquired sideroblastic anemia and the multiple tubular anomalies corresponding to the syndrome initially described by Fanconi has not been reported to date. The underlying mechanism is not understood, but, taken separately, these two anomalies commonly present heme metabolic anomalies in the mitochondria. We hypothesize that this syndrome could represent the clinical expression of a mitochondrial cytopathology. PMID- 8719551 TI - Plasma concentration of human adrenomedullin in patients on hemodialysis. AB - To investigate a possible pathophysiological role of human adrenomedullin (AM), we measured the plasma concentration of immunoreactive-AM (ir-AM) in 38 patients with end-stage renal disease (ESRD) on hemodialysis (HD) and 38 healthy subjects (age and sex matched). In addition, plasma ir-AM was characterized by a reverse phase high performance liquid chromatography. The mean value (+/- SEM) of plasma AM in the patients before HD (10.1 +/- 0.67 fmol/ml) was markedly higher than that in the control group (2.9 +/- 0.13 fmol/ml, p < 0.001), but plasma AM levels were not altered by HD. There was a significant correlation between plasma AM levels and mean blood pressure (MBP) in a group of subjects including both patients before HD and healthy subjects (p < 0.01). In chromatographic study, the major peak of ir-AM in the plasma from patients on HD, as well as healthy subjects, emerged at an elution time identical to that of synthetic AM, indicating that the active form of AM was present in the circulating blood. The secretion of AM seemed to be increased in response to the conditions elicited by ESRD such as hypervolemia and/or hypertension, and reduced renal excretion of the peptide may also contribute to its high plasma level. PMID- 8719552 TI - Gamma-heavy chain deposition disease showing nodular glomerulosclerosis. AB - We describe a 35-year-old woman who had nodular glomerulosclerosis associated with deposition of fragmented gamma (gamma 1)-heavy chains. She presented with edema of lower legs, mild proteinuria, and hematuria. Laboratory examination revealed hypocomplementemia, and a small amount of monoclonal IgG-lambda (lambda) in the blood. Renal biopsy disclosed prominent nodular expansion of the mesangium. Ultrastructurally, the nodules were composed of electron dense deposits and fibrillar structures. An immunofluorescent study showed depositions of gamma-heavy chains and C3 in the central portion of nodules and capillary walls, whereas kappa (kappa)-, lambda-light chains, and Fab fragments of the heavy chain were negative. The accumulation of collagen I, IV, V, and VI was demonstrated in the mesangium. Western blot analysis of serum protein disclosed fragmented gamma-heavy chains that did not combine with light chains. Glomerular nodular lesions, thus, can occur in heavy chain deposition disease, as in light chain deposition disease. Fragmented gamma-heavy chains may also induce hypocomplementemia by the activation of the complement pathway. PMID- 8719553 TI - Inappropriate secretion of antidiuretic hormone (SIADH) in a patient with pleurisy related to adult onset Still's disease. AB - The syndrome of inappropriate secretion of antidiuretic hormone (ADH) or SIADH has been so far reported in various disorders. However, to our knowledge, no case of SIADH associated with adult onset Still's disease (AOSD) has been described in the literature. We report on a patient who developed inappropriate secretion of antidiuretic hormone (SIADH) during the course of adult onset Still's disease (AOSD). The patient developed SIADH associated with pleuritis and improved when pleuritis subsided during the course of steroid therapy. Thus, it seems suggest that SIADH was induced by AOSD which is considered to be related with vasculitis. PMID- 8719554 TI - Successful treatment of Aeromonas hydrophila peritoneal dialysis-associated peritonitis using intermittent intraperitoneal gentamicin. PMID- 8719555 TI - Low protein catabolic rate in diabetics in spite of adequate dialysis. PMID- 8719556 TI - Effect of recombinant human erythropoietin on interleukin-1 beta and interleukin 6 in patients on maintenance hemodialysis. PMID- 8719557 TI - Interpreting postural changes in creatinine clearance in patients with renal disease. PMID- 8719558 TI - Does proteinuria down modulate serum erythropoietin? PMID- 8719559 TI - Nocturnal enuresis at the molecular level. PMID- 8719560 TI - Nocturnal enuresis is linked to a specific gene. PMID- 8719561 TI - The normal infant bladder. PMID- 8719562 TI - Bladder dysfunction in children. PMID- 8719563 TI - Is dysco-ordinated voiding in children an hereditary disorder? PMID- 8719564 TI - Is small bladder capacity a cause of enuresis? PMID- 8719565 TI - Technical aspects of assessing bladder function in children. PMID- 8719566 TI - Micturition habits and bladder capacity in normal children and in patients with desmopressin-resistant enuresis. PMID- 8719567 TI - The relationship between the bladder, the kidneys and the CNS. PMID- 8719568 TI - Sleep patterns in children with nocturnal enuresis. PMID- 8719569 TI - Sleep and arousal in adolescents and adults with nocturnal enuresis. PMID- 8719570 TI - Fluid-provoked enuresis-like episodes in healthy children. PMID- 8719571 TI - Hormonal control of sodium and water excretion in obstructive sleep apnoea. PMID- 8719572 TI - Age-related changes in the circadian control of urine output. PMID- 8719573 TI - Polyuric and non-polyuric bedwetting--pathogenic differences in nocturnal enuresis. PMID- 8719574 TI - Night-time polyuria and nocturia. PMID- 8719575 TI - Behavioural treatment for enuresis. PMID- 8719576 TI - SWEET, the Swedish Enuresis Trial. PMID- 8719577 TI - Desmopressin tablet treatment in nocturnal enuresis. PMID- 8719578 TI - Treatment of nocturnal enuresis with the bell-and-pad system. PMID- 8719580 TI - Predictors of response to desmopressin in children and adolescents with monosymptomatic nocturnal enuresis. PMID- 8719579 TI - Why is desmopressin sometimes ineffective at curing bedwetting? PMID- 8719581 TI - Enuresis: psychological and psychiatric aspects. PMID- 8719582 TI - Predicting treatment outcome with conditioning alarms. PMID- 8719583 TI - Influence of culture duration and ciliogenesis on the relationship between ciliary beat frequency and temperature in nasal epithelial cells. AB - Human nasal epithelial cells from excised mucosal specimens were cultured directly in suspension and sequentially on monolayer and in suspension. Ciliary beat frequency (CBF) was measured by fast Fourier transform analysis of computerized microscopic photometry. In biopsy material CBF increased in an approximately linear fashion at 0.6 Hz/degree C between 20 degrees C and 35 degrees C. Above 35 degrees C the increase was lower and was 0.25 Hz/degree C between 40 degrees C and 44 degrees C. CBF increased more rapidly in suspension culture between 25 and 35 degrees C (1 Hz/degree C) but reached a plateau at approximately 40 degrees C and decreased with further temperature elevation. Up to 44 degrees C all changes were reversible, while irreversible slowing and deterioration occurred above 45 degrees C. Values found after 3 weeks' initial suspension culture were similar to those after 6 weeks' sequential monolayer suspension culture. After 3 weeks of ciliogenesis in sequential suspension culture, all values up to 41 degrees C were statistically significantly higher than those under the other conditions. Ciliary activity was maintained and expressed in culture. CBF was higher than in biopsy material and a reversible decrease was observed at high temperature. PMID- 8719584 TI - A multicenter clinical study of the efficacy and tolerability of azelastine nasal spray in the treatment of seasonal allergic rhinitis: a comparison with oral cetirizine. AB - A new topically administered intranasal antiallergic drug, azelastine, was investigated in a large randomized multicenter study that compared it with oral cetirizine from the aspects of efficacy and safety. Patients were treated for 14 days, and efficacy was assessed on days 7 and 14 by means of an investigator rating scale measuring the severity of eight nasal and ocular symptoms of seasonal rhinitis. In addition, patients recorded the extent of individual symptoms on a visual analogue scale (VAS). Tolerability was assessed on the basis of adverse events reported. Data from a total of 129 patients were included in the analysis of drug efficacy. Treatment groups had significant reductions in the investigators' total symptom score during treatment. These reductions were 47% and 55% for azelastine and cetirizine, respectively, at day 7 and 61% and 67% at day 14. There were no differences between the two groups whether they were analyzed overall or separately for nasal and ocular symptoms. Patients' daily VAS scores showed a significantly better resolution of nasal stuffiness and rhinorrhea in the azelastine-treated group than in the cetirizine-treated group. There were no differences for any other symptom. Adverse events were reported by 12 patients in the azelastine group and 20 patients in the cetirizine group. Drowsiness was the only frequently occurring event and this was reported by 9 patients in the cetirizine group and 2 patients in the azelastine group (P = 0.003). PMID- 8719585 TI - The effect of preoperative bone conduction threshold levels on the results of stapes surgery. AB - The effect of preoperative bone conduction (BC) threshold levels on the results of stapedectomy was evaluated in 378 adult patients with otosclerosis. The mean follow-up period was 8.8 years. Both short-term and long-term postoperative hearing levels were found to be significantly dependent on preoperative BC levels. Only 2 of 31 patients (7%) with preoperative BC thresholds > 40 dB had postoperative hearing levels of 30 dB or better. However, hearing still improved by > or = 21 dB as often in patients with elevated preoperative BC thresholds as in patients with normal BC thresholds. These findings support the use of stapes surgery being worthwhile even in patients with highly elevated BC thresholds. PMID- 8719586 TI - Calcitonin gene-related peptide in vitro stimulation of ciliary beat in human upper respiratory cilia. AB - Calcitonin gene-related peptide (CGRP) is one of the neuropeptides that is released in the human nasal mucosa after trigeminal nerve stimulation. We investigated the in vitro effect of CGRP on the ciliary beat frequency (CBF) of human upper respiratory cilia. Ciliated epithelium of freshly removed adenoidal tissues was used in all experiments. CBF was measured using a computerized photo electric method. The results showed that CGRP exerted a significant dose dependent stimulation on CBF in concentrations of 10(-9)-10(-6) M (n = 10), with a maximum of 23%. Our findings indicate that CGRP in vivo may help to protect the airway epithelium by directly stimulating ciliary beat. PMID- 8719587 TI - Neutral endopeptidase activity and concentration of sensory neuropeptide in the human nasal mucosa. AB - Human nasal mucosa biopsy samples were studied by biochemical and histological methods to determine whether the concentration of calcitonin gene-related peptide like immunoreactivity (CGRP-LI) as a marker of sensory nerves was dependent on the activity of neutral endopeptidase-like enzyme (NEP-LE). Mucosal samples from the middle turbinate were obtained from 32 patients undergoing functional endoscopic nasal surgery for non-allergic chronic rhinosinusitis. The degree of symptoms related to nasal obstruction, rhinorrhea and headaches was recorded. The number of inflammatory cells in each biopsy sample was evaluated by conventional histopathological examination. CGRP-LI was measured by radioimmunoassay. The activity of NEP-LE was evaluated in vitro using [3H] Leu5-enkephalin as substrate. A good correlation was observed between increased concentrations of CGRP, abundant inflammatory cells and the intensity of symptoms (R2 = 0.80). A low activity of NEP-LE was associated with a high concentration of both inflammatory cells and CGRP, suggesting that NEP-LE activity was reduced during inflammation. These observations further support the hypothesis that reduced degradation of sensory neuropeptides could be involved in the pathophysiological mechanisms of non-specific chronic rhinosinusitis. PMID- 8719588 TI - Morphological studies on the pathogenesis of Reinke's edema. AB - Light microscopy of vocal cord mucosa in patients with Reinke's edema revealed highly ramified fissured spaces in the subepithelial tissue that were generally lined with flat cells. The ultrastructure of the parietal cells resembled fibroblasts whose cytoplasmic extensions overlapped in two to three layers in some places. Cell contacts were not observed. Neither electron microscopy nor immunohistochemical testing with antibody against laminin demonstrated a basal membrane. It was possible to distinguish between light and dark cells in the specimens examined. The cytoplasm of the light cells contained intermediate filaments, mitochondria, lysosomes, coated vesicles, caveolae and broad cisternae of rough endoplasmic reticulum. The dark cells were more numerous and typically exhibited a well-developed endoplasmic reticulum and free ribosomes. The parietal cells showed no immunoreaction against human vascular endothelial cells. Immunohistochemical demonstration of mesenchymal intermediate filaments using antibody against vimentin yielded a positive reaction for some of the cells in the walls of the crevices and subepithelial tissue. It was also possible to demonstrate a few cells with monoclonal antibody against macrophages (KiM6). These findings contradict the concept of lymphatic distension in cases of Reinke's edema. Since the parietal cells seen resembled synoviocytes in their structure and immunohistochemical reactions, findings indicate that the hollow spaces of Reinke's edema develop like neobursae from mechanical strain. PMID- 8719589 TI - Experimental tissue expansion induces changes in expression of procollagen I and III messenger RNA. AB - Experimental tissue expansion was performed in nine dogs following placement of subcutaneous silicone balloons. The balloon expander was then filled with 300 ml saline immediately after implantation. Duration of expansion varied from 3 days to 124 days. Unaffected skin and skin over an empty expander served as control tissue. Dermal procollagen I and procollagen III gene expression in response to tissue expansion was investigated by dot-blot analysis using digoxigenin-labeled RNA probes complementary to either human procollagen-alpha 1(I) mRNA or procollagen-alpha 1(III) mRNA. Cross-hybridization of human probes with canine procollagen mRNA was demonstrated by Northern blot analysis. In response to the trauma of surgery, procollagen I and III mRNA transcriptions were found to be decreased significantly within the first few days after implantation. After 9 days of expansion, increased levels of procollagen I mRNA were found, while after 16 days increased levels of procollagen III mRNA were evident. The present study is the first to demonstrate changes in dermal collagen gene expression as a reaction to tissue expansion. PMID- 8719590 TI - Regulatory peptides in the human soft palate. AB - The distributions of vasoactive intestinal polypeptide, peptide, histidine methionine, helospectin, neuropeptide Y and its C-flanking peptide, substance P and calcitonin gene-related peptide were studied in the human soft palate using immunocytochemical techniques. Peptide-containing nerve fibers were found to form a dense network around glandular acini, excretory ducts and blood vessels, as well as beneath and within the epithelium. Chromogranin A, bombesin-flanking peptide and calcitonin gene-related peptide immunoreactivities were detected in endocrine-like cells located in excretory ducts. PMID- 8719591 TI - Basaloid squamous cell carcinoma of the larynx. AB - Basaloid squamous cell carcinoma is a variant of squamous cell carcinoma that was first described in 1986. In the English-language literature, only 26 cases of this infrequent histopathological entity have been reported in the larynx. Tumors have an aggressive biological behavior, justifying combined surgical treatment with radiotherapy. Two new cases are presented and the literature reviewed. PMID- 8719592 TI - The effect of tonsillectomy on neutrophil chemotaxis in adults with chronic tonsillitis. AB - Neutrophil random motility and chemotactic responsiveness were determined in 17 healthy adults and 19 adult patients with chronic tonsillitis before and 10 days after tonsillectomy, using a modified Boyden chamber method. The mean chemotactic index values of the healthy controls were significantly higher when compared with the chronic tonsillitis patients before tonsillectomy. The postoperative mean chemotactic values were increased, and there was a statistically significant difference between the mean chemotactic index of neutrophils in preoperative and postoperative patients. The authors conclude that this change in host defense mechanism may be the reason for frequent attacks of acute tonsillitis in these patients. PMID- 8719594 TI - The three-dimensional fibrillar arrangement of the basilar membrane in the mouse cochlea. AB - The three-dimensional fibrillar arrangement of the basilar membrane in the mouse cochlea was studied using scanning electron microscopy. Fibrils of the basilar membrane were exposed by removing cellular elements of the cochlea using a sodium hydroxide maceration technique. The arrangement of fibrils in the basilar membrane was different between the pars arcuata and pars pectinata. In the pars arcuata, fibrils were arranged in radial and spiral direction, showing a woven pattern. In the pars pectinata, most of the fibrils ran in the radial direction. These findings suggest that the vibration pattern of the pars arcuata and pars pectinata is different when the basilar membrane vibrates. PMID- 8719593 TI - An external approach without tracheotomy for submucosal cordectomy with laterofixation for bilateral abductor paralysis of the vocal folds. AB - There are numerous surgical procedures now available to manage bilateral abductor paralyses of the vocal folds. These procedures have various success rates but mostly do not offer reliable and predictable postoperative results, and usually require tracheotomy. The technique described in this report is based on a trial study to obtain a safe airway for an affected patient and avoid tracheotomy, if possible. Submucosal cordectomy was used with lateral fixation of one vocal fold and preservation of the arytenoid. The procedure was performed by using continuous intravenous anesthesia, which does not require intubation of the patient's airway. Seven patients were operated with this technique with excellent postoperative results. The glottal airway was largely improved in all patients, with only minor temporary complications. PMID- 8719595 TI - Nucleolar organizer regions in glottic carcinomas: comparison of DNA cytofluorometry and clinicopathological analysis. AB - The significance of nucleolar organizer regions (NORs) and nuclear DNA content in 73 glottic carcinomas was assessed for proliferative activity and tumor progression. NORs stained with silver colloid were counted, and nuclear DNA content was assayed by cytofluorometry. The cytofluorometric study demonstrated that the percentage of tumors with aneuploidy tended to increase as histological differentiation decreased. Survival rates of patients with diploid and aneuploid tumors were not significantly different. AgNOR staining revealed that mean AgNOR numbers rose as histological differentiation of tumors decreased. Moreover, as T and N categories and stages showed advancing malignancy, mean AgNOR numbers tended to rise. However, there was no significant difference in survival rates between tumors with low and with high AgNOR counts. These studies indicate that while AgNOR staining is better than DNA cytofluorometry for determining histological differentiation of glottic carcinoma, neither is of prognostic value at the present time. PMID- 8719596 TI - Appearance of free radicals in the guinea pig inner ear after noise-induced acoustic trauma. AB - The emergence of superoxide anion radicals (O2-) in the guinea pig inner ear following acoustic trauma was investigated by histochemical methods. Five minutes after exposure to sound at 120-125 dB SPL for 3 h, an O2- reaction product was detected in the cochlea along the luminal membrane of the marginal cells of the stria vascularis. This reaction product could not be found at 30 min, but reappeared at 2 h. The first appearance of O2- is not explainable by our studies, but the second appearance may be related to recirculation of strial blood flow after blood flow stasis. The present observations raise the possibility that free radicals are produced in the inner ear after acoustic trauma and lead to inner ear damage. PMID- 8719597 TI - Inhibitory effects of selenium, vitamin A and butylated hydroxytoluene on in vitro growth of human tongue cancer cells. AB - The effects of vitamin A, selenium and butylated hydroxytoluene (BHT) on the growth of a human tongue cancer cell line were examined in monolayer cell cultures. A colony-forming assay showed a 50% reduction in the survival rate of the cell line at a concentration of 2.6 micrograms/ml selenium, 60 micrograms/ml vitamin A and/or 38 micrograms/ml BHT. Relatively low concentrations of selenium markedly inhibited glucose consumption. Flow cytometric analysis with both fluorescein-isothiocyanate-labelled bromodeoxyuridine monoclonal antibody and propidium iodide demonstrated an increase in S-phase fractions 1 day after the addition of selenium, increased G0/G1 phase fractions in the presence of vitamin A and increased G2-M phase fractions when BHT was added. These results suggest that these compounds inhibit DNA synthesis of in vitro human tongue cancer cells by different mechanisms. PMID- 8719599 TI - A new outlook for anti-oxidant defense system. PMID- 8719598 TI - Detection of human papillomavirus type 57 in a case of inverted nasal papillomatosis in Japan. AB - Human papillomavirus (HPV) type 57 DNA was detected in recurrent nasal inverted papillomatosis, in a 60-year-old Japanese male, using Southern blot hybridization. HPV types 6, 11, 16 and 18 were not detectable in the papilloma. Previous studies have shown that HPV types 6, 11, 16 and 18 were mainly found in nasal papillomatosis in other laboratories but the actual detection rate of virus was low. We speculate that this low detection rate may be due to the lack of a method for detecting HPV type 57. PMID- 8719600 TI - Oxygen radical detoxification processes during aging: the functional importance of melatonin. AB - That free radical destruction of macromolecules is a basis of aging and age related diseases has considerable experimental support. Melatonin, a hormone produced in organisms as diverse as algae and humans, is believed to have evolved coincident with aerobic metabolism. In all organisms melatonin is produced primarily during the daily period of darkness, with only small amounts being synthesized during the day. In mammals including man, melatonin is produced by and secreted from the pineal gland during the night; however, the night-time production of melatonin falls markedly with aging such that in senescent animals a night-time melatonin rise is barely measurable. This may be significant in terms of aging in the light of recent observations which show that melatonin is a highly efficient free radical scavenger and antioxidant both in vitro and in vivo. In vitro, melatonin has been shown to directly scavenge both the hydroxyl and peroxyl radical, and it does so more efficiently than other known antioxidants. Furthermore, melatonin greatly potentiates the efficiency of previously-discovered endogenous and exogenous antioxidants. In vivo, both physiological and pharmacological levels of melatonin reportedly counteract the devastatingly destructive actions of free radical generating chemicals. For example, melatonin effectively combats DNA damage in rats given massive doses of the chemical carcinogen safrole, and the indole overcomes much of the genomic damage inflicted by ionizing radiation. Also, lipid peroxidation induced by either paraquat, bacterial lipopolysaccharide or H2O2 is highly significantly reduced by concurrent melatonin administration. Finally, cataracts produced in newborn rats by the depletion of the endogenous antioxidant glutathione are prevented by melatonin. These findings provide evidence that melatonin is operative in the cell nucleus, in the aqueous cytosol and in lipid-rich cellular membranes as an antioxidant. Considering this, the loss of this potent antioxidant during aging may be consequential in terms of cellular and organismal aging as well as the onset of age-related diseases. These experimental results from a variety of sources suggest that a more determined approach to the study of melatonin as an anti-aging factor is warranted. PMID- 8719601 TI - Study of aging rat tail collagen using atomic force microscope. AB - Collagen structure of young and old rats was examined by using atomic force microscope (AFM) images. Rat tail tendons of eight and twenty-four month-old Wistar rats were digested enzymatically (pepsin), and allowed to refibrillate for 24 hours at 37 degrees C. The samples were examined using a Nanoscope III (Digital Instruments, Santa Barbara, CA, U.S.A.) with a J scanning head and a 200 microns silicon nitride cantilever. The study was performed in air and without filters. The AFM inspection of refibrillated collagen produced images showing long fibrils with relatively homogeneous heights and widths, characterized by clear banding with a periodic interval (D band) of 67 nm. With respect to collagen extracted from young rats, collagen extracted from old rats revealed fibrils exhibiting the same band interval, but with lower widths and heights. Furthermore, the depth of gap between two overlaps showed a higher mean value in the aged rats. These data are consistent with biochemical reports of collagen modifications during aging; we suggest that post-synthetic reactions might be responsible for this as they interfere with the refibrillation process and also modify the three-dimensional structure of fibrils. PMID- 8719602 TI - A method for scoring the pain map of the McGill Pain Questionnaire for use in epidemiologic studies. AB - Identifying and quantifying the location of pain may be important for understanding specific functional impairments in elderly populations. The purpose of the present analysis was two-fold: first, to describe the reliability of a scoring method for the McGill Pain Map (MPM), and second, to validate the method of scoring the MPM as a tool for assessing areas of body pain in an epidemiologic study. In interviews performed at the subjects' homes, 411 community dwelling Mexican-American and non-Hispanic white subjects aged 65-74 from the San Antonio Longitudinal Study of Aging (SALSA) were asked to describe the location of their pain on the map of the human body included in the McGill Pain Questionnaire. The location of pain was scored by overlaying the survey figures with a MPM template divided into 36 anatomical areas. Inter- and intra-rater agreement among three raters was measured by calculating a kappa statistic for each of the body areas, and an intraclass correlation coefficient for the total number of painful areas (NPA). Internal validity was measured by Spearman's rho between the NPA and the Present Pain Index (PPI) and Pain Rating Index (PRI) of the McGill Pain Questionnaire, and external validity by correlation between NPA and the Perceived Health (PH), Amount of Bodily Pain (APB), and Pain Interference with Work (PIW) items of the Medical Outcomes Study, and the Perceived Physical Health (PPH) question of the San Antonio Heart Study. Average inter-rater agreement for individual MPM areas was 0.92 +/- 0.01, and average agreement for NPA was 0.96 +/ 0.01. Intra-rater agreement for individual areas averaged 0.94 +/- 0.01, and for NPA = 0.99 +/- 0.001. Pain in one or more areas was present in 47.7% of the subjects. For the whole sample, correlations between NPA and the validation indices were: PPI (0.91), PRI (0.89), PH (0.25), ABP (0.64), PIW (0.49), and PPH (0.20). Among the 196 subjects with pain, correlations were: PPI (0.34), PRI (0.34), PH (0.19), ABP (0.21), PIW (0.38), and PPH (0.19)-p < 0.01 for all correlations. In conclusion, we have developed a reliable method of scoring the MPM and have shown evidence of its validity in a community-based sample of elderly subjects. Patterns of painful body areas may be associated with specific diseases and functional impairments. PMID- 8719603 TI - Responses of people of different ages to health problems. AB - There is relatively little information available about the response of people of different ages, particularly elderly people, to symptoms of ill health. This background information is essential for altering inappropriate responses, for such responses may cause long delays or even a complete lack of treatment for people who may be responsive to such treatment. This paper indicates that older people may not choose as wide a range of different health care services as younger people. They tend to concentrate their attention on calling the general practitioner or, in the case of foot trouble, the chiropodist, without recourse to other services. In particular, they tend to avoid using the emergency services, even where appropriate, and prefer to refer problems to the general practitioners, rather than the practice or community-based nurses. It will be necessary to help older people to understand the wider roles that nurses in particular are undertaking if primary care is to be seen as the central focus of health services in future. There is need for more health education by general practitioners for elderly people. This is especially important since the role of general practitioners is likely to change as the emphasis on primary care increases. PMID- 8719604 TI - Different kinds of services and informal support in the daily life activities in a community-based elderly population. AB - As part of the well-known population study of 70-year olds in Gothenburg, 1239 persons at the age of 70 were invited to participate in an extensive investigation and Inter-Vention program of the Elderly in Gothenburg (IVEG). The probands were examined at the ages of 70, 73 and 76 years. The purpose of this study is to describe different kinds of services and informal support in the daily life activities among three large groups of people in their seventies. A cross-sectional study design was chosen in order to get as large groups as possible. A home call was made by an experienced occupational therapist, who interviewed and observed the probands' performance in the daily life activities. The probands and their relatives were asked about the extent of different services and informal support. Personal and home assistance care given by relatives almost doubled between the age of 70 and 76 years. Help organized by the Social Welfare Services only increased marginally. Of the personal activities of daily living (ADL), getting in/out of a bathtub was the activity that caused most problems. The most common walking aid was a stick or a crutch, while use of a wheelchair was quite uncommon. Between the age of 70 and 76 years, the use of the Handicap Transport Service more than doubled. This study shows that at the age of about 73 years the elderly should be offered formal support, such as the supply of assistive devices. What is noteworthy is that the help from relatives and friends increased much more than the help from the Social Welfare Services between the age of 70 and 76 years. At present, relatives and friends assist the elderly in their homes to a large extent, and frequently. Relatives are doing an important job for their elderly and for society, and should be given every possible kind of public support and relief in order to have the strength to continue. PMID- 8719605 TI - Effect of age on marrow macrophage number and function. AB - Employing flow cytometry and a monoclonal antibody against the murine macrophage antigen, Mac-1, we found a significant increase in the number of marrow macrophages in aged mice. This was reflected as significant increase with age in the number of alpha-naphthyl acetate esterase positive cells, as well as in colony forming unit-macrophage (CFU-M) progenitor cells. Macrophages from the marrow of old mice generated significantly less tumor necrosis factor alpha (TNF alpha) than did macrophages from young mice, either spontaneously or when activated by granulocyte-macrophage colony stimulating factor (GM-CSF). Furthermore, conditioned medium (CM) derived from either marrow or peritoneal macrophages of old mice caused less suppression of burst forming unit-erythroid (BFU-E) colony growth than did CM obtained from young mice. Aging, therefore, is associated with an increase in the number of marrow macrophages that have an impaired ability to generate or release cytokines. The increase in macrophage number may reflect a compensation for their reduced function. Altered macrophage number and function may contribute to the age-related decline in hematopoietic reserve capacity. PMID- 8719606 TI - Lipoprotein(a) plasma levels and apo(a) isoforms are not associated with longevity or disability in a sample of Italian octo-nonagenarians. Associazione Medica Sabin. AB - Cardiovascular diseases are the leading cause of disability and mortality in western countries. Lipoprotein(a) [Lp(a)] is now considered an independent risk factor for atherosclerosis, and might consequently be related to longevity and/or disability. In the context of a study on metabolic and anthropometric parameters in a sample of Italian octo-nonagenarians, Lp(a) and apo(a) isoforms were evaluated. One-hundred and fifty Italian octo-nonagenarians were classified as free-living or disabled, according to Katz's index, and compared to 91 healthy control adults. All the study subjects were recruited from a valley (Val Vibrata valley) near Teramo, in the central part of Italy. The median Lp(a) concentration of the whole group was 17 mg/dL (range 1-161 mg/dL), which is much higher than the values observed in Caucasian populations. No differences were detected between the octo-nonagenarian group (median 16 mg/dL, range 1-126 mg/dL) and the control group (median 19.5 mg/dL, range 1-161 mg/dL), nor between the free-living and the disabled groups. Apo(a) isoforms were similarly distributed among free living, disabled and control subjects. While our findings suggest that Lp(a) plasma levels and apo(a) isoforms are not factors associated with longevity or disability, we cannot exclude that the low incidence of other major risk factors for atherosclerosis in our free-living octo-nonagenarians hampered the full expression of the lipoprotein(a) atherogenic potential, and thus allowed the achievement of a very old age in a good healthy status, even in carriers of high Lp(a) levels or small apo(a) isoforms. PMID- 8719607 TI - A physical fitness model of older adults. AB - Physical fitness for young people is viewed as a multidimensional construct, in that it consists of specific components such as strength, mobility, balance, flexibility, and stamina. This study examined whether this structure underlying physical fitness is also relevant to older adults. A 10-item performance test, which was assumed to assess six components of physical fitness, was administered to 69 healthy volunteers ranging in age from 61 to 83 years. A covariance structure model was applied to the test data: the second-order factor was Physical Fitness, and the first-order factors were Strength, Walking, Balance, Flexibility, Stamina, and Manual Speed which were assumed to be measured based on the ten observed variables. Goodness-of-fit index (GFI) of the model was acceptable (GFI = 0.93). While four factors relating to basic motor performances (Strength, Walking, Balance, and Manual Speed) had loadings more than 0.62 to Physical Fitness, Flexibility and Stamina had less than 0.35. It was suggested for elderly people that strength, mobility, balance, and speed components of physical fitness were highly correlated and explainable by a single factor, while flexibility and cardiorespiratory endurance were to be measured by use of specific measures. PMID- 8719608 TI - The set test as a diagnostic tool in elderly outpatients with suspected dementia. AB - The set test was validated in 138 outpatients (mean age 74 years, 65% women) against a global clinical assessment using the DSM-III-R criteria for dementia. The test proved to have low sensitivity even at cutpoints close to a full score, and 12 of 72 patients diagnosed with dementia had a full score. We were unable to confirm the hypothesis that the set test can distinguish between vascular and Alzheimer's type of dementia. Used in its original form, the set test does not seem to be an appropriate diagnostic tool in the setting of a memory clinic. PMID- 8719609 TI - Prolonging life in elderly people: a worthwhile goal of medical care. AB - The very elderly (over 80 years) have a relatively high level of disability and dependency, and this has been reported to be a greater problem in women. It has, therefore, been proposed that there may be little to gain from prolonging life of the very elderly, especially very elderly women. However, there is very little evidence to suggest that increasing survival leads to a higher prevalence of disability at a given age. Preventive measures that prolong life appear to postpone both morbidity and mortality. Moreover, there is little reliable evidence that an elderly woman is more dependent than a man of the same age. It is also suggested that there is a 'cap' on survival and, therefore, we should concentrate on preventing morbidity and compress it into the last few years of life. We consider whether or not the elderly are approaching a 'cap' on survival and think that they are not. We conclude that preventive medicine in the elderly should aim to prolong survival and prevent morbidity. It is uncertain whether morbidity can be prevented more than mortality or vice versa. The concept of compressing morbidity at the end of life is an attractive concept, but data are lacking to support this idea. PMID- 8719610 TI - Androgen receptor-like immunoreactivity in the Brazilian opossum brain and pituitary: distribution and effects of castration and testosterone replacement in the adult male. AB - Androgens are involved in a variety of centrally mediated functions after binding to their intracellular receptors. In the present report, we have employed the androgen receptor antibody, PG-21, and indirect immunohistochemistry to examine the distribution of cells containing androgen receptor-like immunoreactivity (AR IR) in the intact adult male Brazilian opossum brain and pituitary. Additional adult males were castrated to examine the effects of withdrawal of circulating androgens and testosterone replacement on AR-IR. Immunoblots and immunohistochemical controls demonstrated that the androgen receptor in the opossum brain and peripheral tissues are of a similar molecular mass as to has been reported for the rat. Cells containing AR-IR were widely distributed throughout the brain of intact adult males. The highest number of immunoreactive cells were present in the dorsal and ventral nuclei of the lateral septum, medial division of the bed nucleus of the stria terminalis, medial preoptic area, median preoptic nucleus, nucleus of the lateral olfactory tubercle, central amygdaloid nucleus, anterior cortical amygdaloid nucleus, posterior amygdaloid nucleus, subiculum, ventromedial hypothalamic nucleus, arcuate-median eminence region, and ventral premammillary nucleus. The anterior pituitary gland also contained a high number of cells containing AR-IR. The general distribution of AR-IR both in the brain and anterior pituitary gland resembled that reported for other mammalian species. Castration of the adult males four days prior to perfusion eliminated androgen receptor immunostaining throughout the brain except for a few lightly immunostained cells in the ventral nucleus of the lateral septum and stria terminalis. Androgen receptor immunostaining was decreased in the anterior pituitary gland following castration and became cytoplasmic. Testosterone administration 2 h before perfusion restored AR-IR both in the brain and anterior pituitary gland. These data suggested that immunohistochemical detection of bound (nuclear) androgen receptors as seen with PG-21 antibody in the brain and anterior pituitary gland of the opossum is dependent upon circulating androgens. Further, the wide distribution and similarity in localization of androgen receptors in the opossum brain and anterior pituitary gland to that of other species suggests that androgen receptors might be involved in similar functions in the opossum as has been reported for other species. PMID- 8719611 TI - Sodium current in periglomerular cells of frog olfactory bulb in vitro. AB - Kinetic properties of the sodium current in periglomerular (PG) cells were investigated by applying whole-cell patch-clamp techniques to thin slices of the frog olfactory bulb. Eight of the cells were intracellularly stained with Lucifer Yellow for precise identification. Under current-clamp conditions PG cells showed rich spontaneous activity at rest. Na current was isolated from other current contributions by equimolar substitution of K+ with Cs+ in the intracellular solution to prevent K-currents, and 100 microM Cd2+ in the external solution to block Ca-current. Depolarisations beyond -40 mV activated a fast transient TTX sensitive inward current. Once activated, INa declined exponentially to zero following a single exponential. The underlying conductance showed a sigmoidal activation between -40 and +30 mV, with half activation at -17.4 mV and a maximal value of 9.7 nS per neurone. The steady-state inactivation was complete at -30 mV and completely removed at -90 mV, with a midpoint at -56 mV. The activation process could be adequately described by third order kinetics, with time constants ranging from 260 microseconds at -20 mV to 70 microseconds at +50 mV. PMID- 8719612 TI - LTP varies across the estrous cycle: enhanced synaptic plasticity in proestrus rats. AB - Previous studies have shown that the number of dendritic spines and synapses in hippocampal CA1 stratum radiatum decreases more than 30% between the proestrus (high estrogen) and estrus (low estrogen) phases of the rat estrous cycle [10,27]. In the present study, we investigated whether hippocampal synaptic plasticity, as measured by long-term potentiation (LTP), might also vary across the estrous cycle of the female rat. Male rats, and female rats at each phase of the estrous cycle were tested in either the morning or afternoon. There were no significant group differences in the pre-LTP I/O curves. However, females examined during the afternoon of proestrus, the phase during which prior studies indicate synapse number to be highest, demonstrated the greatest degree of potentiation. Diestrus, proestrus and estrus females tested in the morning demonstrated similar amounts of potentiation. There were also significant differences in post-LTP I/O curves between the afternoon proestrus females and males tested in the afternoon. These results suggest that gonadal hormones, interacting with the time of day, may regulate neural processes underlying learning and memory. PMID- 8719613 TI - Discrimination and study of rhythmical brain activities in the alpha band: a neuromagnetic frequency responsiveness test. AB - In this paper we propose a fast procedure--stimulation paradigm, data analysis, data presentation--that permits the study of frequency selective enhancements or suppressions of the cortical responses in selected frequency windows. These responses can be used to classify the underlying activity. This novel procedure combines, in one single test, multichannel detection, finely frequency scanned stimulation, a paradigm that permits to study the effect of the stimulation on the underlying activity (during the non-stimulated periods intermingled with the burst of stimuli) and a compact presentation to appreciate the whole rate/frequency/topographical dependency of the spectral responses. In particular we applied this procedure to non-invasively study for the first time the frequency responsiveness within the extended alpha band (6 Hz to 14 Hz) under separate visual and somatosensory stimulation. The test proved robust and the responses quite stationary. We discriminated the lower and higher alpha band through their different topographical and frequency responsiveness features and suggested them to be the same sub-bands recently discriminated via cognitive experiments on the basis of their functional correlates. We measured a rate dependent alpha suppression (both in the somatosensory and visual modality) that is suggested to be linked to the decoding of the stimulus rate. A slight decrease in the frequency of the spontaneous activity following stimulations at any rate is suggested to be connected to attentional load. We stressed the importance of our simple sensorial discrimination of those activities already evidenced on the basis of their functional correlates, as well as the possible clinical uses of the test: on epilepsy, Alzheimer's disease (recently described to have effects on the responsiveness of the cortex to sensory stimuli at different frequencies) as well as other disabling pathologies. PMID- 8719614 TI - Time-related roles of excitatory amino acid receptors during persistent noxiously evoked responses of rat dorsal horn neurones. AB - The responses of convergent dorsal horn neurones to peripheral injection of formalin (5% formaldehyde, 50 microliters volume) were recorded extracellularly in the halothane anaesthetized rat. The control response of dorsal horn neurones to formalin was biphasic, with a first phase from 0-10 min and the second inflammatory phase from 10-60 min. Pre-administered intrathecal CNQX (5, 50 and 500 micrograms), 5 min before formalin injection, significantly reduced both the first phase (40 +/- 22, 52 +/- 20 and 40 +/- 28% inhibition, respectively, P < 0.05) and the second phase of the formalin response (40 +/- 20% inhibition, P < 0.05, 93 +/- 4% inhibition, P < 0.0001 and 65 +/- 17% inhibition, P < 0.05, respectively). Post-administered CNQX, administered 5 min after the peripheral injection of formalin, was less efficacious, as compared to pre-administered CNQX, at reducing the second phase of the formalin response. The lowest dose of post-administered CNQX (5 micrograms) facilitated the second phase of the response (47 +/- 19% facilitation, P < 0.05), and the higher dose (50 micrograms) produced smaller inhibitions of the response (42 +/- 10% inhibition, P < 0.05) than those observed with pre-administration of the same dose. However, the highest dose of CNQX (500 micrograms) studied produced similar inhibitions of the second phase of the formalin response, irrespective of the timing of administration. Intrathecal administration of 7-chlorokynurenate (7CK, 0.25-2.5 micrograms), a functional antagonist at the glycine site of the NMDA receptor, did not alter the first phase of the formalin response. The second phase of the formalin response was significantly inhibited, and to a similar extent, by both pre- and post-administration of 2.5 micrograms of 7CK (67 +/- 10% and 56 +/- 7% inhibition respectively, P < 0.05 for both). Overall, our results clearly demonstrate differential time-related roles of different transmitter systems in the induction and maintenance of inflammatory evoked persistent pain responses, and such events may become increasingly relevant to the control of pain in the clinic. PMID- 8719615 TI - Electrophysiology and GABA-immunocytochemistry in the vestibular nuclei of normal (C57BL/6J) and Leaner mutant mice. AB - Neurological disorders of genetic origin that lead to distinct disarrangements of the cerebellar wiring and cause a specific motor behaviour are likely to differentially influence the response properties and activity of postsynaptic cerebellar target neurons in the deep cerebellar (DCN) and vestibular nuclei (VN). Comparative electrophysiological and morphological analyses of these neurons in different mutants may increase our understanding of the physiological consequences of cell damage to the cerebellum and help to elucidate the relationships between histopathology and severeness of motor impairment. The Leaner mutation removes GABAergic inhibitory input to the VN predominantly originating from Purkinje cells (PC) located in the anterior lobe of the cerebellum and causes extremely severe motor disability when compared to other cerebellar mutants. In the present study the electrophysiological properties of naturally stimulated (sinusoidal head and body rotation) neurons in the VN of Leaner mutants and their corresponding wild-types (C57BL/6J) were investigated. Neuronal activity of VN single units in Leaner mutants is significantly increased over that of wild-types (frequency range 0.2-0.6 Hz) and more pronounced for type I neurons than for type II. Phase relationships and spontaneous activity are similar at these frequencies in both groups. To elucidate the degree of GABAergic input loss of VN-neurons, quantitative morphometric and numerical analyses of GABA-immunopositive synaptic boutons in the lateral VN of Leaner mutants were performed in addition and revealed significantly smaller terminals and a massive decrease (80%) in Leaner mutant terminal numbers compared to controls. In the context of the findings recently obtained in Weaver and Purkinje cell degeneration (PCD) mutants, the results in Leaner suggest that the loss of inhibition due to the PC degeneration in the anterior vermis leads to a differential enhancement of type I and type II target neuron activity in the VN. The overall activity in Leaner is intermediate between PCD (no increase in activity) and Weaver (strong increase of type I but no increase of type II). GABA immunocytochemistry supports the idea that in Leaner the lost GABAergic PC innervation of the lateral VN has not been replaced by surviving PCs, which is in contrast to Weaver where sprouting of GABAergic terminals in this nucleus was observed. Substitution of lost cerebellar inhibition by non-cerebellar sources, as suggested in the case of PCD-mutants, is uncertain in Leaner due to the increased activity of type I target neurons. These conditions may contribute, among others, to the severe motor disturbances in Leaner. PMID- 8719616 TI - Carbamazepine induction of apoptosis in cultured cerebellar neurons: effects of N methyl-D-aspartate, aurintricarboxylic acid and cycloheximide. AB - We have previously demonstrated that carbamazepine (CBZ) at concentrations above the therapeutic range is toxic to cultured cerebellar granule cells. Here, we ask whether the effect of CBZ involves neuronal apoptosis or necrosis. Treatment of cultured cerebellar granule cells with CBZ for 3 days resulted in a concentration dependent fragmentation of DNA revealed as a laddered pattern in agarose gel electrophoresis, a phenomenon characteristic of apoptosis. Pretreatment of cells with N-methyl-D-aspartate (NMDA) blocked CBZ-induced DNA fragmentation and neurotoxicity as assayed by loss of mitochondrial activity with MTT or by [3H]ouabain binding to Na+/K(+)-ATPase. Aurintricarboxylic acid (ATA), a polyanionic dye, also markedly suppressed DNA fragmentation and cell death detected by morphological examination. A considerable level of DNA ladder formation was detected in untreated cells and this basal DNA fragmentation was also blocked by NMDA and ATA. Moreover, NMDA and ATA prevented CBZ-induced chromatin condensation as revealed by DNA binding with the fluorescent dye Hoechst 33258. Pretreatment of cells with cycloheximide, a protein synthesis inhibitor, prevented CBZ-induced cell death detected morphologically and attenuated CBZ-induced neurotoxicity assessed by mitochondrial activity and [3H]ouabain binding assays. Taken together, our results suggest that CBZ-induces death of cerebellar granule cells by an apoptotic process that is sensitive to NMDA, ATA and cycloheximide. PMID- 8719617 TI - Growth-promoting effects of endothelin-1 and endothelin-3 on chick embryos. AB - Chick embryos were treated with single or multiple doses of 0-250 pmol of endothelin-1 (ET-1) or endothelin-3 (ET-3) and their effects on development were examined in embryos ranging from 7 to 14 days in age. The overall sizes of the head, trunk, beak or wings were increased (5-14%) in ET-treated embryos. Additionally, the lengths or widths of the telencephalic or optic lobes and the diencephalon in the embryonic brains were also increased (4-14%). These effects were observed when the embryos were treated with ET for three consecutive days beginning on day 2 of incubation or from day 1-5. Similar results were obtained by a single injection given on the 5th day of incubation, but not on day 1 or 3. Morphological studies suggested that ET-1 and ET-3 promoted maturation rather than non-specific growth in size. Histological studies revealed that ET-treated brain tissues were more cellular and organized in appearance than control brains. There was no increased incidence of vascular abnormalities or structural malformations subsequent to ET-treatment. These results suggest that ET has growth factor-like effects and that treatment on and around the 5th day of incubation seemed to be most effective for the early phase in brain development in chick embryos. PMID- 8719618 TI - InsP3-gated ion channels in rat olfactory cilia membrane. AB - InsP3-gated channels present in isolated rat olfactory cilia membranes were studied by recording current fluctuations in cilia membranes fused onto phospholipid bilayers formed at the tip of a patch pipette. We found that InsP3 (1.25 to 30 microM) induced current fluctuations in 18 of 157 otherwise silent bilayers. The ion channels opened by InsP3 could be classified into two groups that differed in terms of conductance and kinetics. One channel, which had a conductance of 37 pS, displayed two current levels, and the larger single channel currents were associated with longer open dwell times. The other channel displayed a larger conductance (103-184 pS). Both types of channels displayed reversal potentials near zero millivolts, consistent with nonspecific cation channels. These experiments suggest that mammalian olfactory neurons possess two types of ciliary InsP3-gated channels. PMID- 8719619 TI - Inhibition of potassium-stimulated acetylcholine release from rat brain cortical slices by two high-affinity analogs of vesamicol. AB - In this work, we investigated the effects of two structural analogs of the drug vesamicol, which inhibits the vesicular acetylcholine (ACh) transport, on the potassium-stimulated release of ACh from rat brain cortical slices. These vesamicol analogs, 4-aminobenzovesamicol (ABV) and (trans)-cyclohexovesamicol (transDec), were almost as potent as vesamicol in inhibiting the evoked release of ACh from cortex slices. Similar to vesamicol, the presence of these analogues inhibited the ability of ACh newly-synthesized from [3H]choline to become releasable. However, vesamicol's action was reversible, while ABV and transDec caused a persistent block of this [3H]ACh release. In addition, vesamicol did not affect the release of pre-stored [3H]ACh, but ABV and transDec partially inhibited the release of [3H]ACh in this condition, suggesting that the two latter drugs may alter some of the steps posterior to the entry of [3H]ACh into synaptic vesicles. The rank order of potency for these drugs to reduce ACh release (vesamicol = transDec > ABV) is close to the rank order for inhibition of ACh vesicular transport (transDec > vesamicol > ABV), but is completely different from the order of affinities of these drugs for the vesamicol receptor (ABV > transDec > > vesamicol). These results suggest that although these two vesamicol analogs are able to block ACh release due to their effects on the vesicular transport system, they may have other unexpected actions not shared by vesamicol. PMID- 8719620 TI - Sleep deprivation in rats with preoptic/anterior hypothalamic lesions. AB - Chronic total sleep deprivation (TSD) of rats by the disk-over-water method reliably produces initial increases and subsequent decreases in waking intraperitoneal (Tip) and hypothalamic (Thy) temperatures, progressive increases in energy expenditure, skin lesions on the tail and plantar surfaces, debilitated appearance, and eventual death. We investigated the possible role of the preoptic/anterior hypothalamus (POAH) in the mediation of the TSD effects by comparing these effects in POAH-lesioned and unlesioned rats. Bilateral POAH lesions sufficient in size to impair homeothermic responses to changes in ambient temperature did not produce TSD-like temperature changes under baseline ambient temperatures of 28-29 degrees C, implying that the thermoregulatory changes produced by TSD do not result from impairment of the lesioned area. However, the possibility remains that the TSD effects are mediated by damage to POAH areas that were not lesioned. During TSD, lesioned and unlesioned rats showed similar progressive increases in energy expenditure, but the lesioned rats showed earlier, steeper, and eventually greater declines in Tip and Thy. This result suggests that in unlesioned rats the POAH may counter-regulate against, and thereby attenuate, the reduction in heat retention caused by TSD. This failure of regulation in lesioned rats is consistent with their impaired response to ambient temperature change and implies that, in unlesioned rats, some POAH thermoregulatory mechanisms continue to function normally during TSD. Lesioned rats did not show the characteristic TSD-induced early increases in Tip and Thy. This result could imply either that heat retention was so compromised that body temperatures did not rise in spite of a TSD-induced increases in thermoregulatory setpoint, or that the setpoint increase in unlesioned rats is POAH-mediated. Notwithstanding the greater Tip and Thy declines in lesioned rats, they survived the TSD procedure longer than the unlesioned rats, thus supporting previous indications that death did not result from hypothermia. PMID- 8719621 TI - Forskolin reduces the activity of the rat muscle embryonic type acetylcholine receptor channel. AB - The action of forskolin (FSK), a stimulator of cAMP-dependent protein kinase (PKA), on nicotinic acetylcholine receptor-(nAChR-) channels was studied on cultured rat muscle fibres. The channel activity was estimated by determining Np, with N, being the number of channels and p, the single channel open probability. In order to elucidate the possible role of PKA in the modulation of nAChRs, FSK (10-50 microM) was added to the bath or to the pipette filling solution in the cell-attached configuration. The first protocol used was to test for indirectly mediated cytosolic effects, the other, for any direct effects of the drug on nAChR-channels. Using both experimental protocols, no effects on the duration of single-channel openings or conductance were observed, while channel activity was significantly reduced. In particular, FSK 10 microM caused a reduction of Np only when applied to the non-patch membrane. FSK at higher concentrations, produced a more marked decrease of Np when present in the recording pipette. The present work provides evidence that the channel activity of muscle embryonic-type nAChRs can be influenced by a direct action of FSK, and is also significantly reduced by an indirectly-mediated cytosolic mechanism triggered by FSK. PMID- 8719622 TI - Impaired myelinated fiber regeneration following freeze-injury in rats with streptozotocin-induced diabetes: involvement of the polyol pathway. AB - This study examined the effects of streptozotocin-induced diabetes mellitus and aldose reductase inhibitor (ZD5522) treatment on myelinated fiber regeneration in rats. After 2 months of diabetes, with or without ZD5522 treatment (10 mg.kg 1.day-1) from induction, sciatic nerve degeneration was initiated by a punctate lesion using a liquid N2 cooled probe. Regeneration was studied over a subsequent 14-day period using in vitro electrophysiological techniques. There was a 21.4% (P < 0.001) deficit in the maximum fiber regeneration distance in diabetic rats, 14 days postlesion. This was partially (64.9%, P < 0.01) prevented by aldose reductase inhibitor treatment, the resultant regeneration distance being not significantly different from that of age-matched nondiabetic control rats. The regeneration rate, assessed from data collected 4, 9 and 14 days postlesion, was 23.7% (P < 0.001) reduced by diabetes and ZD5522 treatment provided 73.1% protection (P < 0.01). We conclude that polyol pathway activity is involved in impaired regeneration in experimental diabetes, potential pathophysiological mechanisms involving a reduction in neurotrophic support and impaired endoneurial blood supply. PMID- 8719623 TI - Distribution of two molecular forms of gonadotropin-releasing hormone (GnRH) in the central nervous system of the frog Rana ridibunda. AB - Two molecular forms of gonadotropin-releasing hormone (GnRH) have been recently characterized in the brain of the frog Rana ridibunda i.e. mammalian GnRH (mGnRH) and chicken GnRH-II (cGnRH-II). Using highly specific antisera against each form of GnRH, we have investigated the distribution of these two neuropeptides in the frog brain by the indirect immunofluorescence and the peroxidase-antiperoxidase techniques. mGnRH-immunoreactive cell bodies were restricted to a well defined region corresponding to the septal-anterior preoptic area. mGnRH-containing fibers projected through the ventral diencephalon and ended in the median eminence. In contrast, cGnRH-II-immunoreactive structures were widely distributed in the frog brain. In the telencephalon cGnRH-II-positive elements formed a ventromedial column extending from the olfactory bulb to the septal area, a pathway which corresponds to the terminal nerve. A dense accumulation of cGnRH-II immunoreactive cell bodies was also found in the septal-anterior preoptic area; these neurons sent processes towards the median eminence via the hypothalamus. Double immunostaining revealed that, in this area, mGnRH- and cGnRH-II-like immunoreactivity co-existed in the same neurons. In the mid-diencephalon, numerous cGnRH-II-immunoreactive perikarya were found, surrounding the third ventricle, in the posterior preoptic and infundibular areas. Many of these neurons sent processes towards the ventricular cavity. More caudally, a dense population of cGnRH-II-immunoreactive perikarya was also observed in the nucleus of the paraventricular organ and the posterior tubercle. Dorsally, the thalamus, the tegmentum, the tectum and the granular layer of the cerebellum were richly innervated by cGnRH-II-positive fibers. In the medulla oblongata, numerous cGnRH II-immunoreactive perikarya were seen in several cranial nerve nuclei. Ventrally, a dense plexus of immunoreactive fibers projected rostrocaudally into the spinal cord. The occurrence of mGnRH- and cGnRH-II-like immunoreactivity in the septal anterior preoptic area and the hypothalamo-pituitary pathway supports the view that both peptides act as hypophysiotropic neurohormones. The widespread distribution of cGnRH-II-immunoreactive elements in the central nervous system of the frog strongly suggests that this peptide may also exert neuromodulator and/or neurotransmitter activities. PMID- 8719624 TI - Adenovirus-mediated expression of a reporter gene in thalamocortical cocultures. AB - Organotypic cocultures of thalamic and cortical explants have recently been used to study the development of the thalamocortical axonal network in the mammalian neocortex. To explore the possibility of genetically manipulating organotypic explants, rat thalamocortical (TC) cocultures were infected with the recombinant adenovirus, Adv/RSV beta gal. Infection of the cortical explants resulted in long term expression (2 weeks) of the reporter gene (beta-galactosidase) with no significant alterations to the structural integrity of the explants. By micro injecting the adenoviruses into cortical explants a significant degree of spatial control over reporter gene expression was obtained. DiI-labeled axonal projections from thalamic explants into infected (n = 116) and control cortical (n = 120) explants were also analyzed. There was no significant difference in the extent or degree of TC ingrowth into infected or control cortical explants. Thalamic explants were also efficiently infected with the Adv/RSV beta gal virus. While the pattern and extent of TC ingrowth from infected thalamic explants was similar to controls, the percentage of viable, infected thalamic explants was decreased. These experiments were necessary precursors for future studies using recombinant adenoviruses and organotypic cocultures. Genetic manipulation of these cocultures should enable the dissection of proteins involved in the development of axonal networks in the mammalian neocortex, using a system amenable to direct manipulation and observation. PMID- 8719625 TI - Source localization of middle latency auditory evoked magnetic fields. AB - The magnetic counterparts of middle latency auditory evoked responses (MLR) were measured for seven normal subjects with a 7-channel de superconducting quantum interference device (SQUID). The source of each component (Na, Pa, Nb and Pb) was estimated and plotted onto the individual magnetic resonance images (MRI). The source of Na, as well as those of Pa, Nb and Pb, was estimated to be in the supratemporal auditory cortex. The positions of Pa, Nb and Pb sources were compared with one another. No significant difference was observed between the positions of Pa and Nb sources. On the other hand, the source of Pb was found to be anterior to the sources of both Pa and Nb. It was suggested that there are more than two separate areas activated in the human auditory cortex during MLR. PMID- 8719626 TI - Decreased cerebral blood flow during acute hyperglycemia. AB - Cerebral blood flow (CBF) decreases during acute hyperglycemia but the mechanism of this change is unknown. The role that plasma osmolality plays in this effect was reexamined in pentobarbital-anesthetized rats using a continuous measure of CBF, laser-Doppler flowmetry. CBF decreased 25% during acute elevation of plasma osmolality induced by intraperitoneal injection of concentrated solutions of glucose or mannitol. In addition there were brief transient increases of CBF with peak magnitude 2-4-times the baseline level that were not accompanied by transient depression of electroencephalographic activity. These transient CBF increases may explain why discontinuous methods of CBF measurement fail to detect flow decreases after mannitol injection. Decreased CBF measured during acute hyperglycemia may be the result of increased plasma osmolality. PMID- 8719627 TI - Tumor necrosis factor-alpha-induced gelatinase B causes delayed opening of the blood-brain barrier: an expanded therapeutic window. AB - Proteolytic damage is a late event in the molecular cascade initiated by brain injury. Earlier, we proposed that matrix metalloproteinases (MMPs) and urokinase type plasminogen activator (uPA) are important in secondary brain injury. We have shown that intracerebral injection of activated 72-kDa type IV collagenase (gelatinase A) opens the blood-brain barrier, and that during hemorrhagic brain injury there is endogenous production of 92-kDa type IV collagenase (gelatinase B) and uPA. Therefore, to study the functional link between proteolytic enzymes and blood-brain barrier damage, we induced MMP expression by infusing tumor necrosis factor-alpha (TNF) intracerebrally in rats. Initially, the effect on capillary permeability of increasing doses of TNF, using [14C]sucrose uptake, was measured. Then, the time-course of the capillary permeability change was studied at 4, 16, 24 and 72 h. Expression of MMP and uPA was measured by zymography at 24 h after TNF injection and compared to saline-injected controls. A dose-dependent increase in capillary permeability was seen 24 h after TNF injection. Maximal uptake of [14C]sucrose occurred at 24 h compared to saline-injected controls (P < 0.05). Zymography showed production of gelatinase B, which was significantly greater than in saline-injected controls at 24 h (P < 0.05). Batimastat, a synthetic inhibitor to metalloproteinases, reduced sucrose uptake at 24 h (P < 0.0001), and was effective even when given 6 h after TNF (P < 0.01). Thus, gelatinase B is the intermediate substance linking TNF to modulation of capillary permeability. Agents that interfere with transcription of proteolytic enzymes or block their action may reduce delayed capillary injury, extending the therapeutic window. PMID- 8719628 TI - D1 dopamine receptors influence Fos immunoreactivity in the globus pallidus and subthalamic nucleus of intact and nigrostriatal-lesioned rats. AB - Studies of the globus pallidus (GP) and subthalamic nucleus (STN) have emphasized the role of D2 dopamine receptors, although effects of D1 receptor activation on GP firing rate and STN metabolism have been reported, especially in rats with nigrostriatal lesions. This study systematically investigated the effects of D1 and D2 receptor activation on the activity of the GP and STN in intact and 6-OHDA lesioned rats using immunostaining for the immediate-early gene Fos. In intact rats, the D1 agonist SKF 38393 (20.0 mg/kg) produced a five-fold potentiation of the GP Fos expression due to the D2 agonist quinpirole produced significant Fos expression. In rats with prior nigrostriatal lesions, SKF 38393 (4.0 or 20.0 mg/kg) increased Fos immunostaining in both the GP and STN, while quinpirole increased it only in the GP. SKF 38393 effects in the GP and STN of nigrostriatal lesioned rats were blocked completely by SCH 23390, and unaffected by eticlopride. These results are a novel demonstration of control of Fos expression by dopaminergic drugs in the STN and by D1 agonists in the GP. PMID- 8719629 TI - Deafferentiation-induced c-fos gene expression in subthalamic nucleus and substantia nigra reticulata is reduced by non-NMDA receptor antagonist. AB - Molecular events underlying the mechanism by which brain injury elicits delayed transneuronal degeneration of neurons remote from the site of initial injury are not well understood. In rats, acute injury of the caudate nucleus (CN) and globus pallidus (GP) by local injection of excitotoxic ibotenic acid (IA) or by transient forebrain ischemia resulted in delayed cell death of neurons in the substantia nigra reticulata (SNr). To elucidate the involvement of glutamate receptor mediated hyperactivity of neurons produced by loss of inhibitory inputs in this delayed degeneration of SNr neurons, the region-specific expression of an immediate early gene, c-fos, and the effect of glutamate receptor antagonists on the c-fos expression were examined by using immunocytochemical and in situ hybridization analysis. Following unilateral IA-injection into the CN and GP, a robust expression of c-fos mRNA and Fos protein was induced specifically in neurons of both subthalamic nucleus (STN) and SNr deafferented by the IA-lesions 36 h after IA-injection. The delayed expression of Fos-protein in SNr neurons lasted for 48 h longer than that in STN neurons. Following unilateral IA injection confined to the CN, an intense but short-term expression of Fos-protein was exhibited only in neurons of the deafferented SNr. c-fos mRNA and Fos protein were not expressed in neurons of the substantia nigra compacta at any time points examined. The induction of c-fos mRNA and Fos protein in neurons of the STN and SNr following IA-lesions of the CN and GP was reduced markedly by non-NMDA receptor antagonist (GYKI52466), but not by NMDA receptor antagonist (MK-801). The region-specific c-fos expression implies that deprivation of inhibitory afferents (disinhibition) due to destruction of presynaptic neurons can induce increased activity of postsynaptic neurons. The effect of GYKI52466 on the c-fos gene expression in neurons of the deafferented STN and SNr suggests that activation of non-NMDA receptors may be involved in a pathophysiological cascade for the transneuronal degeneration of SNr neurons. PMID- 8719630 TI - Vanilloid (capsaicin) receptors in the rat: distribution in the brain, regional differences in the spinal cord, axonal transport to the periphery, and depletion by systemic vanilloid treatment. AB - Vanilloid (capsaicin) receptors were visualized by [3H]resiniferatoxin (RTX) autoradiography in the brain of newborn as well as adult (both control and colchicine-treated) rats. Specific labelling was seen in the brain stem only, in the nucleus of the solitary tract extending into the area postrema and the spinal sensory nucleus of the trigeminal nerve. Also, a strong signal was seen in the dorsal horn, dorsal root, trigeminal and nodose ganglia. Membranes obtained from the cervical, thoracic, and lumbar segments of the spinal cord showed similar affinities for RTX and likewise for capsaicin and capsazepine; maximal receptor density was similar in the cervical and thoracic segments (approximately 70 fmol/mg protein) but was twice as high in the lumbar segment. 24 h after ligation of the vagal or the sciatic nerves, a strong accumulation of specific RTX binding sites was observed mainly proximal to the ligature, implying intraaxonal receptor transport from the nodose and dorsal root ganglia, respectively, to the periphery. Systemic (s.c.) vanilloid treatment depleted specific [3H]RTX binding sites from the brain stem, the sensory (dorsal root as well as trigeminal) ganglia, and the spinal cord. RTX was approximately 200-fold more potent than capsaicin for eliminating vanilloid receptors from the spinal cord. The present results suggest a discrete expression of vanilloid receptors in the brain stem (sensory nuclei); although intrinsic vanilloid receptor-expressing neurons are though to exist in the rat brain, they remain undetected by the present [3H]RTX autoradiography methodology. PMID- 8719631 TI - Liposome-entrapped phenytoin locally suppresses amygdaloid epileptogenic focus created by db-cAMP/EDTA in rats. AB - Status epilepticus was induced in rats by injecting a combination of dibutyryl cAMP (db-cAMP) and ethylenediaminetetraacetic acid (EDTA) into the amygdala (AM), and the effect of phenytoin (PHT), entrapped in liposomes (PHT-L) and given intravenously at 40 mg/kg, on the spiking activity of the AM epileptogenic focus was examined. Electroencephalograms were recorded from the db-cAMP/EDTA-injected AM and the bilateral sensorimotor cortices. One dose of PHT-L, given 30 min after intra-AM db-cAMP, produced immediate and transient seizure suppression, but did not suppress the sequential spiking activity, which lasted for more than 5 h. In contrast, two doses of PHT-L, given 30 and 60 min after intra-AM db-cAMP/EDTA, produced delayed and local suppression of AM discharges, and immediate and transient seizure suppression was also observed. The AM discharges began to be suppressed about 100 min after the second injection of PHT-L injection, with no overt change occurring in cortical spiking activity. This was followed by total seizure suppression about 170 min after the second PHT-L injection. This effect was not observed after one or two injections of PHT alone. When horseradish peroxidase (HRP), to which the blood-brain barrier is impermeable, was entrapped in liposomes (HRP-L) and given intravenously 30 min after intra-AM db-cAMP/EDTA, an accumulation of HRP was found in the db-cAMP/EDTA-injected AM in 2 of the 5 animals tested. With 2 doses of HRP-L given 30 and 60 min after intra-AM db cAMP/EDTA, the local augmentation of HRP in the AM was found in all 5 of the 5 animals tested. Our findings suggest that: (1) the AM epileptogenic focus created by db-cAMP/EDTA has a high affinity for liposomes, and this factor participates in the local suppression of AM discharges by PHT-L, and (2) two injections of PHT L are required for the AM to gather an effective amount of PHT-L. PMID- 8719632 TI - Quantitative autoradiographic analysis of D1 and D2 dopamine receptors in rat brain in early and late pregnancy. AB - This study examined the levels of D1 and D2 dopamine receptors in the rat brain during pregnancy, a physiologically unique and important naturally occurring state. We are particularly interested in changes in the dopamine receptor complement of the brain during pregnancy because these receptors might support some components of the immediate postpartum onset of normal maternal behavior. Quantitative in vitro receptor autoradiography was applied particularly focusing on brain areas that control maternal behavior. The D1 dopamine receptor selective antagonist -3H-SCH23390 and the D2 dopamine receptor selective antagonist [3H]spiperone were used as the ligands. We examined the levels of binding to D1 and D2 dopamine receptors in brains in females on day 2 (early pregnancy) and day 21 (late, but prepartum pregnancy) of pregnancy. In addition, brains from females on diestrus-1 and from males provided reference points to the existing literature. Late in pregnancy females had significantly 18-27% lower levels of binding to D1 dopamine receptors in the lateral striatum, the medial striatum, and the nucleus accumbens when compared to all other groups. Late in pregnancy, females had also significantly 11-25% lower levels of binding to D2 dopamine receptors in the lateral striatum, the anterior striatum, the nucleus accumbens and the olfactory tubercle compared to all other experimental groups. We examined all of the brain regions already established to be important for maternal behavior, and found that dopamine receptor binding changed across pregnancy only in one such region, the nucleus accumbens. Thus pregnancy, perhaps the hormones of pregnancy, reduces the levels of D1 and D2 dopamine receptors in the striatum, and the nucleus accumbens, but not in other brain regions. PMID- 8719633 TI - beta-Phenylethylamine regulation of dopaminergic nigrostriatal cell activity. AB - In the present paper, the action of beta-phenylethylamine on electrophysiological activity of dopaminergic nigrostriatal neurons is described. 10 s after its i.v. injection and during 2-4 min, beta-phenylethylamine decreased the firing rate, the number of spikes within and out of burst and the number of bursts per second of these neurons. This was a dose-related action with statistical differences starting from 1.4 mg/kg for total and out of burst firing rate and from 2.4 mg/kg for within burst firing rate and for the number of bursts per second. The standard deviation and the variation coefficient of inter-spike intervals increased in a dose-related way. The marked effect found after low-dose administration suggests that under physiological conditions endogenous beta phenylethylamine levels regulate the nigrostriatal dopaminergic cell activity. After peripheral low dose administration, beta-phenylethylamine behaves as a dopaminergic agonist with a very fast and brief action. PMID- 8719634 TI - Relationship between frequency of spontaneous bursting and tonotopic position in the developing avian auditory system. AB - Neural activity in the developing brainstem auditory pathway of the chick embryo is dominated by a rhythmic pattern of spontaneous discharge. Neurons in nucleus magnocellularis (NM) and nucleus laminaris (NL), second and third order auditory nuclei, discharge spontaneously in synchronous bursts at periodic intervals. Rhythmic bursting is present as early as embryonic day 14 (E14), shortly after the onset of functional synaptogenesis, and gives way to an adult-like, steady level of firing on E19, two days prior to hatching. In the present experiment, multiple-unit recording techniques were used in E17 and E18 embryos to examine the relationship between rate of rhythmic bursting and tonotopic position in NM and NL. The mean rate of rhythmic bursting ranged from 0.21-0.71 Hz. Bursting rate varied systematically as a function of position, being faster at progressively higher frequency regions of the nuclei at both E17 (r = 0.75) and E18 (r = 0.86). In addition, the rate of bursting at a given location in the nuclei increased during development. The presence of a systematic relationship between the rate of rhythmic bursting and tonotopic location suggests that the spatio-temporal pattern of spontaneous discharges could provide developmental cues for the spatial ordering of auditory projections. PMID- 8719636 TI - Direct projections from the orofacial region of the primary motor cortex to the superior colliculus in the macaque monkey. AB - Employing anterograde axonal tracing combined with intracortical microstimulation, we have revealed in the macaque monkey that the orofacial region of the primary motor cortex sends projection fibers to the deep layers of the superior colliculus. The terminal site of these projection fibers was almost localized to the lateral part of the intermediate gray layer at its rostral level, and spatially segregated from that of projection fibers arising from the frontal eye field. The results indicate the existence of a discrete tectal region related specifically to the control of orofacial movements. PMID- 8719635 TI - Synaptotagmin-like expression in the motor nerve terminals of crayfish. AB - Synaptotagmin-like immunoreactivity was shown to be localized at crayfish neuromuscular junctions by whole mount immunocytochemistry. Synaptotagmin-like immunoreactivity was present in both phasic and tonic excitatory terminals and inhibitory nerve terminals. Immunoblots indicated that the antibody DSYT-2 raised against Drosophila synaptotagmin, labelled proteins at relative molecular weights of 87 kDa and 107 kDa in crayfish ganglion and neuromuscular preparations. PMID- 8719637 TI - Possible branching of myelinated primary afferent fibres in the dorsal root of the rat. AB - A combined physiological and morphological examination of rat dorsal root ganglion cells revealed branching of the central process of neurones with myelinated fibres (conduction velocity > 2 m/s; n = 24). Single shock electrical stimulation of spinal dorsal roots triggered double action potentials (early and late spike) in two dorsal root ganglion cells recorded by intracellular electrodes in the in vitro spinal cord-dorsal root ganglion preparation from 12 20 day-old rats. The action potentials had different stimulus thresholds (lower for the late spike). In one dorsal root ganglion cell the differences in delay between the two spikes inhibited the soma invasion of the second spike, when both action potentials were evoked at resting membrane potential. Depolarization of the soma membrane by DC current, injected through the electrode, recovered the blocked action potential. Membrane depolarization by GABA reduced the threshold for the early spike and changed the pattern of activation. After electrophysiological characterisation, intracellular biotin/avidin staining of the neurone revealed branching of the central axon in the dorsal root. None of the other cells, which responded with single action potentials after dorsal root stimulation showed secondary branching (n = 5). This rare observation shows that differences between the conduction velocities and activation thresholds in branches of individual dorsal root ganglion cell axons may produce block of spike invasion into the soma and perhaps the spinal terminal field of large primary afferents. PMID- 8719638 TI - Age-related changes of oxygen free radical production in the rat brain slice after hypoxia: on-line measurement using enhanced chemiluminescence. AB - Superoxide radical production in brain slices of 12-, 24- and 60-day-old rats was measured online during and after 15 min of hypoxia with lucigenin enhanced chemiluminescence. We found a typical radical burst after reoxygenation which developed with aging and is almost nonexistent in the youngest and most prominent in the oldest group. This cannot be explained by a decreasing tissue superoxide dismutase (SOD) concentration in the brain with aging, since the concentration of the enzyme, determined in the same age groups, increased with age. PMID- 8719639 TI - Bifurcated projections of retinal ganglion cells bilaterally innervate the lateral geniculate nuclei in the cat. AB - Cats were injected with the fluorescent retrograde tracers, Fluoro-Gold (FG) and Evans Blue (EB), into the left and right lateral geniculate nuclei (LGN), respectively. About 4.56% of the ganglion cells in the temporal retina were double-labeled by these dyes. 4.7% of these cells were of the large type, 30.3% were of the medium type, and 65% were classified as cells of the small type. These results indicate that members of all three ganglion cell size classes, mainly those of small type, bilaterally innervate the LGN via axonal bifurcation. PMID- 8719640 TI - The immunoreactive profile at the N-terminal region of A beta 1-39/40 but not A beta 1-42 changes with transition from monomer/dimer to further peptide aggregates. AB - Using site-specific antibodies, we assessed the effect of aggregation of various length forms of A beta on the immunoreactive profile of the peptides. All of the antibodies tested reacted with monomeric/dimeric forms of A beta 1-42 and its further aggregates. However, antibodies directed against the 1-24 region of A beta reacted weakly or not at all with A beta 1-39/40 monomers or dimers, but immunoreactivity was enhanced substantially following peptide incubation and aggregation. These results suggest that the conformation of the N-terminal region of monomeric and dimeric A beta 1-39/40 is different from that of aggregated forms, whereas the longer A beta 1-42 does not significantly change its N terminal conformation during beta-sheet fibril formation. These immunochemical results are consistent with previous structural data, and help to explain the differential effects of A beta 1-39/40 and 1-42 on fibril formation in brain. PMID- 8719641 TI - Oxytocin does not act locally in the hypothalamus to elicit norepinephrine release. AB - An in vitro brain slice preparation was used to determine whether facilitatory effects of systemically administered oxytocin on hypothalamic norepinephrine release result from direct action of the neuropeptide on noradrenergic terminals. When oxytocin was superfused onto hypothalamic slices from ovariectomized control and ovariectomized, hormone-treated female rats, the neuropeptide failed to influence either basal or electrically evoked release of 3H-norepinephrine. Thus, it is likely that oxytocin facilitates hypothalamic norepinephrine release by a peripheral mechanism, perhaps vaginocervical contraction. PMID- 8719642 TI - Reduction in numerical synapse density in chick (Gallus domesticus) dorsal hippocampus following transient cerebral ischaemia. AB - Transient forebrain ischaemia was induced by a two-vessel occlusion method in the domestic chick. One week post-surgery, hippocampal tissue was processed for electron microscopy and synapse density assessed using the disector technique. Hippocampal volume was estimated using image analysis of serial coronal cryostat sections. The density of asymmetric synapses was significantly reduced (27%; P < 0.005) in ischaemic chicks. This appears to be a real reduction as hippocampal volume was not significantly decreased. PMID- 8719643 TI - Mechanisms of paresthesiae, dysesthesiae, and hyperesthesiae: role of Na+ channel heterogeneity. AB - Paresthesiae, dysesthesiae, and hyperesthesiae ('positive symptoms') result from ectopic nerve impulses secondary to inappropriate membrane excitability which develops in the setting of chronic sensory axonal injury. The molecular changes in the membranes of dorsal root ganglion neurons which underlie ectopic impulse generation as a result of chronic axonal injury are unknown. Preliminary evidence has suggested that voltage-dependent Na+ channels are one of the participants in the production of ectopic impulses, but the precise form of their participation remains to be determined. The present paper reviews normal sensory anatomy and Na+ channel physiology, as well as clinical syndromes heralded by positive sensations and what is so far known about the cellular and molecular mechanisms underlying them. Properties of two distinct populations of Na+ channels native to the DRG neurons which give rise to cutaneous afferents are described. The biophysical properties of each population of Na+ channels must be tuned with respect to the other in order to cooperate in the generation of action potential activity underlying normal sensory function. A novel hypothesis is put forth suggesting that chronic axonal injury leads to intraneuronal heterogeneity of the populations of Na+ channels in cutaneous afferents, as revealed by their characteristic properties. This may result in one population of Na+ channels activating the other, leading to membrane instability, and possibly to ectopic impulse generation. PMID- 8719644 TI - Morphological and histochemical study of nonhemiplegic muscle in acute stroke patients manifesting respiratory failure. AB - There is little information on skeletal muscle changes in patients with acute stroke. We performed morphological and histochemical examinations of nonhemiplegic sternothyroid muscles biopsied at the time of tracheostomy from 13 patients with acute stroke manifesting acute respiratory failure. Degenerating and regenerating fibers were observed in all 13 specimens. Following characteristic myopathic changes suggestive of mitochondrial abnormalities were also demonstrated in a majority of patients. Namely, ragged-red fibers, focal increase in NADH-TR activity in subsarcolemmal areas and increases in acid phosphate activity were found. The changes were similar but extremely slight in control patients with acute respiratory failure due to causes other than stroke and were absent in the other control patients with adenomatous thyroid tumor. The severity and extent of the histopathological changes in the muscle fibers in patients with acute stroke were closely correlated with the duration of hypoxemia but not with such items as type of stroke, site of cerebral lesion, consciousness level, days of biopsy after the stroke, clinical outcome, levels of serum creatine kinase, myoglobin and PaO2. This acute nonhemiplegic muscle involvement was considered to be a very common complication in severe stroke patients. PMID- 8719645 TI - Late-onset epileptic seizures in patients with leukoaraiosis: a positron emission tomographic study. AB - Leukoaraiosis, found on computed tomographic scans of the brain, is suspected to be of ischemic origin and is frequently associated with progressive decline of cognitive functions in elderly persons. Some of them also develop late-onset epilepsy. The present positron emission tomographic study investigates if these seizures in mentally nonaffected patients with leukoaraiosis are related to an underlying ischemic process. Patients with leukoaraiosis and late-onset seizures have a more important decline of regional blood flow and oxygen consumption in the cortical areas, compared to normal age-matched controls, to patients with cryptogenic symptomatic late-onset epilepsy and to patients with a similar degree of leukoaraiosis but without epilepsy. The present study suggests that both the late-onset seizures and the leukoaraiosis in patients with, at that time, no obvious mental deterioration are premonitory signs of an encephalopathy of unknown origin, possibly leading to cognitive decline. PMID- 8719646 TI - Parietal occipital edema in hypertensive encephalopathy: a pathogenic mechanism. AB - Eight patients with hypertensive encephalopathy from diverse etiologies developed cerebral edema in the vertebrobasilar distribution which resolved after blood pressure was lowered. Parietal occipital edema is a recognized feature of hypertensive encephalopathy. The explanation for this regional pathological variation in hypertensive encephalopathy remains undefined. Some evidence suggests that sympathetic innervation of the anterior cerebral vasculature may be protective, and conversely, the relative lack of sympathetic innervation in the vertebrobasilar vasculature may predispose the parietal occipital region to the development of cerebral edema in hypertensive encephalopathy. PMID- 8719647 TI - Botulinum toxin A in non-dystonic tremors. AB - The present pilot study evaluated the effect of botulinum toxin A on primarily non-dystonic tremors using accelerometry in a single-blind, placebo-controlled design. Resting, postural, intention, or head tremor were assessed before and approximately 1 month after intramuscular saline and botulinum toxin A (25-50 U) respectively. Half of the patients showed > or = 30% placebo effect. Tremor in 10 of 17 patients (60%) studied improved further after botulinum toxin A (range 30 95%), exceeding the placebo effect by > or = 30%. Nine patients demonstrated clinically significant focal weakness in the extensor muscles after botulinum toxin A which interfered with fine movements. Patients were subdivided into PD like and ET-like tremor(s). Both groups experienced large placebo effects for resting tremor, with little or no further improvement after botulinum toxin A. The improvement in postural tremor after botulinum toxin A, of 40% in the PD-like and 57% in the ET-like groups, however, was approximately twice that of placebo. In conclusion, botulinum toxin A exerts a modest tremorlytic effect, however the dose, and its distribution over the sites injected, need to be optimised to minimise focal weakness. PMID- 8719648 TI - Respiratory arrest: a complication of Arnold-Chiari malformation in adults. AB - We describe the cases of 2 adult patients who developed respiratory arrest and were found to have Arnold-Chiari malformation on magnetic resonance imaging. Following posterior fossa decompression both patients improved, though one of them subsequently died during sleep. Imaging of the craniocervical junction should be part of the workup of patients with unexplained apneic episodes. PMID- 8719649 TI - Alien hand syndrome of the dominant hand and ideomotor apraxia of the nondominant hand. AB - Following an extended infarction in the territory of the left anterior cerebral artery including the anterior corpus callosum, a 66-year-old right-handed man exhibited an alien hand syndrome on the right side and an ideomotor apraxia of the left hand. In addition to other typical symptoms of frontal lobe lesions, he presented with an interesting difference in the manifestation of perseveration in either hand. In the right hand, the perseveration was part of the alien hand syndrome, whereas in the left hand perseveration showed in the inability to switch promptly from one movement to another on instruction during praxis testing. PMID- 8719650 TI - Urinary retention and space-occupying lesions of the frontal cortex. AB - The authors present 2 cases in which urinary retention was related to supratentorial cortical lesions. These cases support the presence and function of a supratentorial center involved in control of the voiding reflex loop and indicate that lesions affecting this center can result in urinary retention rather than the syndrome of incontinence, urgency, and increased urinary frequency that is generally attributed to dysfunction of this system. In such cases, a clinical presentation of urinary retention in the absence of associated remarkable neurological deficits renders clinical evidence as a localizing sign. PMID- 8719651 TI - Predicting recovery from common whiplash. AB - The present study aimed to establish criteria for delayed recovery after whiplash injury using a 1-year prospective cohort study. 117 whiplash patients referred from primary care and 16 whiplash patients recruited from the insurance company. At baseline all patients underwent neurological, radiological and psychosocial factors assessment and testing of personality traits, well-being, cognitive ability and cognitive functioning. According to the logistic regression in patients referred from primary care the following initial variables are in significant relationship with poor outcome at 1 year: impaired neck movement, history of pretraumatic headache, history of head trauma, higher age, initial neck pain intensity, initial headache intensity, nervousness score, neuroticism score and test score on focused attention. Employing these variables, correct prediction of outcome at 1 year was found in 88% of patients recruited from the insurance company. Authors conclude that a comprehensive assessment of whiplash patients early after trauma enables physicians to identify patients at risk of delayed recovery. PMID- 8719652 TI - Acute transverse myelitis caused by herpes simplex virus. PMID- 8719653 TI - Peripheral neuropathy: an unrecognized feature of hereditary vitreoretinal degeneration? PMID- 8719654 TI - The alien grasp reflex. PMID- 8719655 TI - Cervical epidural abscess due to actinomycosis: demonstration by MRI. PMID- 8719656 TI - Evidence of axonal reinnervation in two arthropod-borne viral encephalomyelitis patients. PMID- 8719657 TI - The incidence of cerebrovascular insult in the city of Split, Croatia, during the 1991/1992 war. PMID- 8719658 TI - Human monocytes lacking the membrane-bound form of the bacterial lipopolysaccharide (LPS) receptor CD14 can mount an LPS-induced oxidative burst response mediated by a soluble form of CD14. AB - Monocytes and macrophages express a glycosyl phosphatidylinositol (GPI)-anchored lipopolysaccharide (LPS) receptor on the cell surface which enables them to detect minute amounts of LPS released from Gram-negative bacteria. A soluble form of CD14 is also found free in serum, though its physiological function is unknown. the interaction of LPS with CD14 on the monocyte surface leads to an activation of the cells which is manifested in the sudden release of reactive oxygen species, a process referred to as an oxidative burst. In patients suffering from the condition known as paroxysmal nocturnal haemoglobinuria (PNH), the synthesis of GPI anchors is blocked in haematopoietic cells which are therefore unable to express GPI-linked proteins on their surface. In severe cases, over 90% of monocytes lack membrane-bound CD14, though normal levels of the soluble form of the receptor-sCD14-are found in the serum. Despite this lack of membrane-bound CD14, monocytes from PNH patients can respond to low concentrations of LPS. Here we show that the LPS-induced oxidative burst of these PNH monocytes requires a component present in serum. The serum-dependent activation can be inhibited by monoclonal antibodies to CD14, can be removed from the serum by passage over a matrix to which an anti-CD14 antibody has been bound, and the depleted serum can be reconstituted by the addition of either purified natural or purified recombinant soluble CD14. We conclude that an LPS-dependent oxidative burst in PNH monocytes can be mediated by soluble CD14. PMID- 8719660 TI - Effects of polar glycopeptidolipids of Mycobacterium chelonae (pGPL-Mc) on granulomacrophage progenitors. AB - Effects of polar glycopeptidolipids of Mycobacterium chelonae (pGPL-Mc) in the in vivo stimulation of haematopoietic growth and differentiation of murine bone marrow and spleen cells was investigated in this study. Progenitors were determined with a quantitative cultural analysis of bone marrow and spleen cells in methylcellulose using rmGM-CSF and rmIL3. Injection of pGPL-Mc produced a significant time-related increase in the number of bone marrow and spleen CFUs. pGPL-Mc treatment, in particular, increased the number of bone marrow and splenic CFU-GMs, CFU-Gs and CFU-Ms during and after three intraperitoneal administrations. The greatest myeloid stimulation of bone marrow CFU-GMs, CFU-Gs and CFU-Ms was observed between days 7 and 14, with maximal values on days 12 and 14. Highly significant stimulation of splenic CFU-GMs, CFU-Gs and CFU-Ms was observed between days 7 and 10 with maximal values on day 10, while the initial stimulation of these progenitors was observed starting from day 1 in bone marrow and day 7 in spleen. These effects of pGPL-Mc were associated with an increase in granulocyte, monocyte and thrombocyte counts in the peripheral blood. Granulocyte and monocyte counts remained high up until day 12, while those of thrombocytes were prolonged until day 18. May-Grunwald-Giemsastained colony samples and differential white blood cell counts demonstrated that the granulocyte population is composed almost entirely of neutrophils. pGPL-Mc is therefore a broad-spectrum haematopoietic growth factor with a highly promising application in the reversal of chemotherapy- and/or radiotherapy-induced myelo-suppression. PMID- 8719659 TI - Bryostatins trigger human polymorphonuclear neutrophil and monocyte oxidative metabolism: association with in vitro antineoplastic activity. AB - Bryostatin-1-but not bryostatin-13-a macrocyclic lactone isolated from the marine bryozoan Bugula neritina, triggered human polymorphonuclear neutrophil (PMN) and monocyte release of reactive oxygen radicals, as measured by the generation of lucigenin chemiluminescence and by the ferricytochrome c reduction assay. The release of oxygen radicals by bryostatins was sensitive to inhibitors of protein kinases, but resistant to the inhibition of phospholipase A2 activity and arachidonic acid metabolism (prior treatment with mepacrine or indomethacin). Comparison of the effect of protein kinase (PK) inhibitors H-8, H-7 and staurosporine on bryostatin-1-induced neutrophil oxygen radical release further suggested a requirement for activation of phospholipid-dependent PKC, but not for cGMP- or cAMP-dependent PK. In cytostatic assays, PMNs treated with bryostatin-1 inhibited the growth of the erythroleukaemic cell line K562 in a concentration dependent manner. These findings suggest that the reported antineoplastic effect of bryostatins may result at least in part from activation of PMNs and monocytes. PMID- 8719661 TI - Similar binding properties for a neutralizing anti-tetanus toxoid human monoclonal antibody and its bacterially expressed Fab. AB - A high-affinity anti-tenanus toxoid (TT) human monoclonal antibody showing neutralizing activity was isolated from a fusion between mouse myeloma and human splenic cells. Fab fragments from this antibody were obtained using a recombinant phage surface-display expression system. The parental antibody and the corresponding Fab had identical immunological activities, including specificity and affinity. These results confirm the feasibility of developing Escherichia coli expression of monoclonal human Fab from hybridoma cells. PMID- 8719662 TI - Chondroitin-4-sulphate (proteoglycan), a receptor for Plasmodium falciparum infected erythrocyte adherence on brain microvascular endothelial cells. AB - Adherence of Plasmodium falciparum parasitized erythrocytes to the microvascular endothelium is mediated by different receptors expressed by endothelial cells. The study of the adherence of P. falciparum-infected erythrocytes to Saimiri monkey brain microvascular endothelial cells revealed the presence of an additional receptor, which was identified and further characterized. This receptor was also found on the surface of primary human lung endothelial cells (HLEC). We developed two mAbs to this receptor which very efficiently blocked the adherence of parasite strains to Saimiri brain endothelial cells (SBEC). The ability of these mAb to bind to SBEC was partially blocked by chondroitin-4 sulphate (CSA). Competitive inhibition assays on adherence of parasitized red blood cells (PRBC) showed that CSA, but not hyaluronic acid, chondroitin-6 sulphate, dermatan sulphate, keratane sulphate, heparan sulphate or chondroitin 4S-disaccharide, was able to almost completely inhibit PRBC adherence. The same effect was obtained with chondroitinase ABC and AC, but not B, hyaluronidase or heparinase. These results strongly suggest that a member of the chondroitin glycosaminoglycan family, CSA, represents an additional receptor used by P. falciparum PRBC to cytoadhere to microvascular endothelial cells. PMID- 8719663 TI - Induction of anaesthesia: one drug or two? PMID- 8719664 TI - Co-induction of anaesthesia: the rationale. AB - Combination therapy with two or more different drugs, with the intention of reaching the same therapeutic goal, was heavily criticized for a long time. However, it is accepted today, especially when advantages over monotherapy can be shown. For the induction of anaesthesia or for long-term sedation in the intensive care unit, combination therapy may offer an improved effect profile, a more balanced ratio of desired versus adverse effects, an improved time-course of effect, simpler treatment requirements or lower costs. Midazolam and propofol have been investigated as potential partners for those two indications. The mechanism of action, pharmacokinetic properties, pharmacological effect, the way in which they interact at the receptor site, the differences in pharmaceutical formulations, the side-effect profiles and economic considerations were compared. Animal experiments and clinical pharmacology studies have shown that midazolam and propofol have synergy with other centrally active drugs. It could be expected that the relationship between desired effects and adverse effects could be improved by skilful use of the synergism between midazolam and propofol. Co induction of anaesthesia and co-administration in long-term sedation can offer improvements in therapeutic situations compared with monotherapy. These improvements are in terms of a more suitable effect profile, a more favourable ratio of desirable effects to side-effects, optimization of the time-course of effects and reduced costs. PMID- 8719665 TI - Intravenous anaesthetic drug interactions: practical applications. AB - The spectrum of effects that constitutes the state of general anaesthesia can be induced by the combined use of drugs. Each drug has a predominant action regarding one of the anaesthesia components, hypnosis, analgesia and amnesia. However, each agent, when used in combination, not only produces its own expected effect, but it can also modify the effect of another agent acting on a different component. For example, an opioid, in addition to its anti-nociceptive effect, can also potentiate the hypnotic effect of a benzodiazepine. Anaesthetists have long recognized these effects but did not quantify them until recently. Pharmacologists have provided us with techniques to measure drug interactions. We have utilized these techniques to demonstrate and quantify significant pharmacological interactions for hypnotic effect with commonly used intravenous agents. The clinical utility of these combinations can now be exploited precisely for the benefit of all our patients. PMID- 8719666 TI - Co-induction of anaesthesia: the cardiac patient. AB - Cardiac patients pose special problems to the anaesthetist because of their underlying disease and the nature of the corrective surgery. Information about new methods of induction of anaesthesia obtained in fit patients may not be applicable directly to patients with heart disease. More suitable are patients undergoing cardioversion. Titrating intravenous induction agents to response elicited appears to be more important than the agent used, although it is possible to inject too slowly with drugs whose offset of action is by distribution. Anaesthetic agents alone are not sufficient to ablate the response to tracheal intubation, skin incision and sternotomy. Balancing induction of anaesthesia with small doses of opioid can obtund the haemodynamic responses. The effects of a drug used solely for induction of anaesthesia are unlikely to be present at the end of 3 or 4 h of surgery. However, this is not the case with agents used to maintain anaesthesia if early extubation after anaesthesia is practised. Reports of anaesthetic techniques for cardiac surgery tend to give total doses used rather than the timing and dose of the constituent agents. At Papworth Hospital, Cambridge, UK, after opioid premedication, midazolam sedation is used during insertion of some, or all, vascular cannulae. Two main techniques then exist. Either an intravenous or volatile anaesthetic agent is started immediately, supplemented by an opioid and muscle relaxant, or anaesthesia is induced with opioid and relaxant and the anaesthetic agent is begun only after transfer to the operating theatre, just before skin preparation. Either way, the end-point of induction of anaesthesia is difficult to discern in heavily premedicated patients with midazolam sedation. PMID- 8719667 TI - Co-induction of anaesthesia: day-case surgery. AB - The term co-induction of anaesthesia has been applied to the use of two or more drugs to induce anaesthesia. The term was introduced in 1986 to describe the unplanned induction of anaesthesia by non-anaesthetically trained personnel practising sedation. A new benzodiazepine was combined with opioids, with synergistic effects, causing unplanned anaesthesia in an unsuitable environment leading to several fatalities. Currently, planned co-induction of anaesthesia is practised by anaesthetists exploiting drug interactions, particularly synergism, principally between midazolam, fentanyl, sufentanil and alfentanil, and propofol. It can produce an improvement in all phases of anaesthesia, including induction, maintenance and recovery. There are advantages in combining midazolam with propofol, thereby reducing the risk of awareness and also the dose of propofol and hence its side-effects and cost. Propofol is the principal intravenous induction agent for day-case anaesthesia. The pre-administration of 0.03 mg kg-1 of midazolam (approximately 2 mg in normal healthy adults) is now being practised widely. Current papers suggest that 2 mg of midazolam administered to an average, otherwise healthy, adult does not compromise recovery, whereas an increase to 5 mg may be expected to delay the possibility of final discharge of such patients by about 20 min. The use of midazolam and propofol with or without either fentanyl or alfentanil is probably the principal technique for the induction of day-case anaesthesia at the present time. A major advantage is that by reducing the dose of propofol there is less chance of the severe bradycardia that is sometimes associated with the combined use of propofol and opioids, although this can be prevented by vagolytic agents. However, the use of opioids increases the incidence of post-operative nausea and vomiting. Another important drug is ketamine, the effects of which are often additive with other drugs. The combination of ketamine and midazolam is an important technique, particularly in the management of critically ill patients. The alpha 2-agonists, e.g. clonidine and dexmedetomidine, may also have a role in this context in the future. This paper presents the current approach to the co-induction of anaesthesia, particularly in relation to the reduced risk of awareness when using midazolam, and the health economics in relation to the potential reduction in the dose and hence cost of propofol. PMID- 8719668 TI - Propofol versus propofol with midazolam for laryngeal mask insertion. AB - We evaluated the addition of midazolam to propofol during induction of anaesthesia by assessing laryngeal mask tolerance, haemodynamic variables, recovery times and cost. Forty patients (ASA grades I-IV) undergoing elective surgery were allocated randomly to receive a standard dose of propofol or a smaller dose of propofol combined with midazolam. A laryngeal mask was inserted and any episodes of coughing or hiccuping during its insertion or removal were recorded. Anaesthesia was maintained with nitrous oxide and enflurane with fentanyl for analgesia. After surgery, recovery times, pain, shivering, nausea, vomiting and analgesic requirements were recorded. The cost of the drugs used was also calculated. No significant differences were detected in any variables, except that patients given propofol needed more morphine in the recovery ward. The average cost of propofol alone was 3.47 pounds per anaesthetic, while the midazolam plus propofol cost was 2.03 pounds. Adding midazolam to propofol allowed a reduced dose of propofol to be used without adverse effects, while reducing the anaesthetic costs. PMID- 8719669 TI - Anaesthetic drugs and bacterial contamination. AB - We investigated the bacterial contamination of commonly used intravenous anaesthetic agents. Ten trainee anaesthetists drew up four syringes each of propofol, midazolam, thiopentone, 0.9% saline and a culture medium control using their normal practice. A set of syringes was cultured at the time of drawing up and at 2, 4 and 8 h afterwards. No anaesthetist washed his or her hands before drawing up the drugs. Six anaesthetists capped the syringes using sheathed needles. Eight syringes were contaminated with bacteria. Coagulase-negative staphylococci were isolated from six syringes (four sheathed and two unsheathed) and an Acinetobacter sp. from two syringes (one sheathed and one unsheathed). No bacteria were cultured from the midazolam syringes. However, two syringes from each of the other solutions were contaminated. This implies that one syringe was contaminated when it was drawn up, at 2 h a further two had detectable contamination, two more were contaminated at 4 h and three more at 8 h. We cannot recommend drawing up drugs at the start of a list for use later in the day. They should be used immediately after drawing up. Basic hand washing before drawing up drugs may reduce contamination. PMID- 8719670 TI - Peripheral corneal infiltrates associated with contact lens wear. AB - PURPOSE: A retrospective study was performed to review the clinical characteristics of peripheral corneal infiltrates in contact lens wearers. METHODS: The charts of all contact lens patients with peripheral corneal infiltrates 1.5 mm or less in size who presented to the office from 1987 to 1994 were reviewed. RESULTS: The epidemiological and clinical characteristics of peripheral corneal infiltrates associated with contact lens wear were reviewed in 52 patients (64 infiltrates). Forty-four patients presented with a single infiltrate, while the remaining 8 patients had multiple infiltrates. While there was no predilection for a specific quadrant of the cornea, when a subgroup of patients who wore extended wear lenses was analyzed, 19 of the 40 infiltrates were located in the superior quadrant. Forty percent of the patients were wearing disposable extended wear contact lenses, 21% were wearing conventional extended wear lenses, 33% were wearing conventional or frequent replacement/disposable daily wear contact lenses and 6% were wearing rigid gas permeable lenses. The majority of patients had minimal conjunctival inflammation, an anterior stromal cellular reaction and minimal anterior chamber activity. A subgroup of 16 patients had corneal cultures of their infiltrates. In this group, 8 of the 16 had positive cultures. All patients had a resolution of the infiltrates without complications and the majority were refitted to daily wear soft or rigid contact lenses. CONCLUSION: Peripheral corneal infiltrates in contact lens wearers appears to be more common in patients wearing extended wear soft contact lenses. While often considered "sterile" in the literature, a significant number have been shown to be culture-positive. The organisms that have been associated with peripheral infiltrates appear to be less "pathogenic" than those that have been reported to be associated with central corneal ulcer. However, it is probably advisable that patients with peripheral corneal ulcers secondary to contact lens wear should be initially treated with topical antibiotics. PMID- 8719671 TI - Functional measurements on the enlarged endothelial cells of corneal transplants. AB - PURPOSE: To measure the morphologic and functional status of the endothelium of long-term corneal transplants. METHODS: We induced hypoxic corneal edema and measured corneal deswelling (expressed as percent recovery per hour, PRPH, over 7 hours) with a modified optical pachometer in 12 corneas of 8 patients transplanted 7-18 years previously and in 21 normal adult control subjects. Fluorophotometry and specular microscopy were also performed on each eye. Results in the 2 groups were compared with a 2-tailed Student T test for means. RESULTS: The transplanted corneas had significantly decreased PRPH, induced swelling, endothelial permeability to fluorescein, and endothelial cell density compared to the controls, despite a similar estimated open-eye steady-state thickness. CONCLUSIONS: The markedly enlarged endothelial cells of long-term corneal transplants have a reduced ability to deturgesce the cornea and decreased permeability to small solutes. These findings could result from a reduction in intercellular space that occurs when the number of cells lining the posterior surface of the cornea is substantially decreased. PMID- 8719672 TI - Corneoscleral block excision of postoperative anterior chamber cysts. AB - PURPOSE: To describe the clinical presentation, surgical treatment and histopathology of three cases of cystic epithelial ingrowth occurring after cataract surgery. METHODS: Three patients developed anterior chamber cysts, two with an associated conjunctival bleb and fistula, following extracapsular cataract and phacoemulsification surgery. The technique of block excision and corneoscleral graft was used in the three patients. RESULTS: There was no evidence of recurrence twenty-six, twenty-three and seven months postoperatively. Visual acuities were 20/30, 20/20-2 and 20/25-2, with mild astigmatism and well controlled intraocular pressure. Histopathology illustrated that conjunctival and corneal epithelium invaded the anterior chamber through a wound defect. CONCLUSION: At this time, block excision with a corneoscleral graft may be the most definitive surgical treatment of cystic epithelial ingrowth. PMID- 8719673 TI - Physiological strategies for emmetropia. AB - PURPOSE: To identify relationships among age-independent ocular biometric variables which contribute to ocular refraction in adult human eyes, and to identify differences in those relationships between emmetropes and ametropes. METHODS: Manifest refraction (DRS methodology), corneal refractive power (keratometry), and axial anterior segment (anterior cornea to posterior lens), vitreous cavity (posterior lens to anterior sclera) and total globe (anterior cornea to anterior sclera) length (A-scan ultrasonography) were determined in 185 unaccommodated right eyes of adult humans aged 18 to 70 years. There were 136 emmetropes with absolute refractive error < or = 2.0 diopter, and 49 ametropes (47 myopes, 2 hyperopes) with absolute refractive error of 2.25-11.0 diopters. RESULTS: Refraction decreased significantly with increasing globe and vitreous cavity length in emmetropes and ametropes. Anterior segment length was also significantly negatively correlated with refraction in emmetropes, but not in ametropes. Corneal refractive power was not correlated with refraction in either group, but decreased significantly with increasing globe length in both, more strongly in emmetropes. Globe and vitreous cavity length were significantly positively correlated in both groups, more strongly in ametropes. Anterior segment length increased significantly with increasing globe length in emmetropes but not in ametropes. Anterior segment length decreased significantly with increasing vitreous cavity length in ametropes but not in emmetropes. CONCLUSION: These findings indicate an "inflatable anterior segment" as well as the classic "inflatable globe" mechanism of emmetropization. This newly described anterior segment mechanism involving increased separation between the cornea and the lens with increasing globe size appears to be absent in adult human myopia. PMID- 8719674 TI - Behind the optic nerve: an inside view of the primate visual system. PMID- 8719675 TI - Surgical alignment prior to six months of age for congenital esotropia. AB - PURPOSE: To complete the first independent and largest multicenter outcome investigation to analyze the motor and functional results of a series of patients surgically aligned prior to age 6 months and followed for a minimum of 4 years. METHODS: Sixteen patients, surgically aligned at an average age of 4.2 months, were examined at an average of 7.1 years to assess their motor and functional outcomes. RESULTS: Motor and sensory tests showed 11 patients to have a small or negligible motor misalignment at near point with both binocular fusion and gross stereopsis ability. A single patient aligned by 3 months of age demonstrated reproducible refined stereoacuity on sensory testing. However, the patients who achieved alignment by 4 or 5 months did not demonstrate any better quality of binocularity than that found in a previously studied group of patients aligned at 6 months. CONCLUSION: Binocularity that includes refined stereoacuity remains an elusive target and a rare outcome for an ophthalmologist treating congenital esotropia, despite very early surgical alignment. PMID- 8719676 TI - Abnormal electroretinogram associated with developmental brain anomalies. AB - PURPOSE: We have encountered abnormal ERGs associated with optic nerve hypoplasia, macular, optic nerve and chorioretinal colobomata and developmental brain anomalies. Brain anomalies include cortical dysgenesis, lissencephaly, porencephaly, cerebellar and corpus callosum hypoplasia. We describe six exemplar cases. METHODS: Scotopic and photopic ERGs adherent to international standards were performed as well as photopic ERGs to long-duration stimuli. CT or MRI studies were also done. The ERGs were compared to age-matched normal control subjects. RESULTS: ERG changes include reduced amplitude b-waves to blue and red stimuli under scotopic testing conditions. Implicit times were often delayed. The photopic responses also showed reduced amplitude a- and b-waves with implicit time delays. The long-duration photopic ERG done in one case shows attenuation of both ON- and OFF-responses. CONCLUSIONS: Common underlying developmental genetic or environmental unifying casualties are speculated to be at fault in causing these cases of associated retinal and brain abnormalities. No single etiology is expected. Multiple potential causes acting early in embryogenesis effecting neuronal induction, migration and differentiation are theorized. These occur at a time when brain and retinal cells are sufficiently undifferentiated to be similarly effected. We call these cases examples of Brain Retina Neuroembryodysgenesis (BRNED). Homeobox and PAX genes with global neuronal developmental influences are gene candidates to unify the observed disruption of brain and retinal cell development. The ERG can provide a valuable clinical addition in understanding and ultimately classifying these disorders. PMID- 8719677 TI - Excimer laser in-situ keratomileusis (LASIK) under a corneal flap for myopia of 2 to 20 D. AB - BACKGROUND: We report the results of a recent technique of keratomileusis for myopia: excimer laser in-situ keratomileusis (LASIK). METHODS: We studied retrospectively 88 eyes of 63 patients that received LASIK with the Chiron Automated Corneal Shaper and the Summit OmniMed excimer laser under a hinged corneal flap without sutures. RESULTS: Mean follow-up was 5.2 months. Mean spherical equivalent of the manifest refraction before surgery was -8.24 diopters (D)(range -2.00 to -20.00 D). Mean spherical equivalent refraction after surgery was +0.22 D (SD, 1.42 D). Of 40 eyes with a baseline refraction from -2.00 to 6.00 D, 25 (63%) had a refraction within +/- 0.50 D and 37 eyes (93%) within +/ 1.00 D. In 29 eyes with baseline refraction of -6.12 to -12.00 D, postoperative refraction was within +/-1.00 D in 19 (65%). In 19 eyes with baseline refraction of -12.10 to -20.00 D postoperative refraction was +/-1.00 D in 8 (43%). Overall, 64 of 88 eyes (72.8%) had a refraction within +/-1.00 D after surgery. Between three weeks and five months after surgery the change in the mean spherical equivalent refraction was -0.61 D in the myopic direction. Uncorrected visual acuity after surgery was 20/20 or better in 31 eyes (36%) and 20/40 or better in 61 eyes (71%). Three eyes (3.6%) lost two lines or more of spectacle corrected visual acuity, two from progressive myopic maculopathy and one from irregular astigmatism. No eyes had vision threatening complications. CONCLUSION: Excimer laser in-situ keratomileusis (LASIK) under a corneal flap can be an effective method of reducing myopia between -2.00 to -20.00 D, with minimal complications. Current surgical algorithms need modification to improve predictability. Stability of refraction after surgery requires further study. PMID- 8719678 TI - Ocular findings in the Hermansky-Pudlak syndrome. AB - BACKGROUND: The Hermansky-Pudlak syndrome (HPS) is defined by the autosomal recessively inherited triad of tyrosinase-positive oculocutaneous albinism, bleeding diathesis and accumulation of ceroid in tissues. Late complications include: interstitial pulmonary fibrosis; inflammatory bowel disease; and renal failure. PATIENTS AND METHODS: We undertook a non-concurrent prospective study of 55 Puerto Rican patients with HPS (age range 1 to 54 yrs; mean = 19.7 yrs). These patients had a comprehensive ocular examination and a systemic evaluation for HPS. RESULTS: Visual acuities ranged from 20/50 to 5/200. All patients had nystagmus. Forty-three patients had strabismus; esotropia was found in 24 patients; exotropia in 18 patients; and one patient had hypertropia. Posterior embryotoxon occurred in 15 patients and Axenfeld anomaly in 4 patients. Iris pigmentation varied from minimal to almost completely normal. Three patients had cataract formation. The retina was typically albinotic with macular hypoplasia. All patients had cutaneous albinism, bleeding diathesis and various systemic manifestations as part of HPS. CONCLUSION: Ocular findings in HPS include reduced visual acuity; congenital nystagmus, strabismus and cataract. Diagnosis of the syndrome ought to be made preoperatively to help minimize the potential complications associated with bleeding diathesis at the time of extraocular muscle and intraocular surgery in patients with HPS. PMID- 8719679 TI - Progressive hemifacial atrophy. A natural history study. AB - PURPOSE: To describe two very different natural history courses in 2 patients with hemifacial atrophy. Progressive hemifacial atrophy (Parry-Romberg syndrome, Romberg syndrome, PHA) is characterized by slowly progressive atrophy, frequently involving only one side of the face, primarily affecting the subcutaneous tissue and fat. The onset usually occurs during the first 2 decades of life. The cause and pathophysiology are unknown. Ophthalmic involvement is common, with progressive enophthalmos a frequent finding. Pupillary disturbances, heterochromia, uveitis, pigmentary disturbances of the ocular fundus, and restrictive strabismus have also been reported. Neurologic findings may be present, but the natural history and progression of ocular findings are often not described in the literature. METHODS: We studied the records and present findings of 2 patients with progressive hemifacial atrophy who were observed in our institution over a 10-year period. RESULTS: Both patients showed progression of ophthalmic findings, primarily on the affected side. One patient has had chronic uveitis with secondary cataract and glaucoma, in addition to retinal pigmentary changes. She also had a third-nerve paresis of the contralateral eye and mild seizure activity. The other patient had mild uveitis, some progression of unilateral retinal pigmentary changes, and a significant increase in hyperopia in the affected eye, in addition to hypotony at age 19 without a clear cause, but with secondary retinal and refractive changes. CONCLUSION: Ocular manifestations of progressive hemifacial atrophy are varied, but can progress from mild visual impairment to blindness. PMID- 8719681 TI - Retinal changes associated with neurofibromatosis 2. AB - INTRODUCTION: Neurofibromatosis (NF) is now known to be more than one disease. NF2, formerly classified as central neurofibromatosis, is characterized by bilateral vestibular schwannomas, previously termed "acoustic neuromas", and is much less common than NF1. Lens opacities at an early age have been described in approximately 85% of NF2 patients. PURPOSE: To determine the frequency of retinal abnormalities in NF2 patients. METHODS: We prospectively examined 15 consecutive patients who met the diagnostic criteria of NF2. RESULTS: We observed an epiretinal membrane in the macular or paramacular area in 12 of 15 patients, and a combined pigment epithelial and retinal hamartoma in the macula of one patient who also had an epiretinal membrane in the macula of the other eye. Additionally, 11 patients had central posterior cortical, subcapsular, or peripheral cortical lens opacities. CONCLUSIONS: Children or young patients with epiretinal membranes, combined pigment epithelial retinal hamartoma, and lens opacities that are not the result of other ocular disorders should have a neurologic evaluation and a careful family history for NF2. PMID- 8719680 TI - Rehabilitation of a child with partial unilateral cryptophthalmos and multiple congenital anomalies. AB - PURPOSE: This paper describes the surgical rehabilitation of a child with craniofacial anomalies, unilateral syndactyly, and partial unilateral cryptophthalmos associated with inferior colobomata of the iris and optic nerve and agenesis of the inferior rectus and inferior oblique muscles. The clinical presentation of cryptophthalmos is described. METHODS: The medical literature since the original description of cryptophthalmos in 1872 was reviewed to define patterns of inheritance and the incidence of associated anomalies. RESULTS: Including this patient, 149 case reports of cryptophthalmos were identified. In two families transmission from parent to child suggests dominant inheritance. None of the five dominant cases had any other anomalies, and all had bilateral complete cryptophthalmos. The incidence of cryptophthalmos in the remaining families is consistent with autosomal recessive inheritance. This group includes patients with bilateral, unilateral, and partial cryptophthalmos. Other anomalies are common, including those of the ear and nose, limbs, genitourinary system, and mouth and palate. Mortality in the perinatal period is associated with renal agenesis, laryngeal atresia, and pulmonary hypoplasia. CONCLUSIONS: Cryptophthalmos is a rare congenital anomaly with two patterns of inheritance. PMID- 8719682 TI - Risk factors for growth and metastasis of small choroidal melanocytic lesions. AB - PURPOSE: To investigate the clinical features that predict growth and metastasis of an unselected group of small melanocytic choroidal tumors. METHODS: A retrospective review was performed on 1329 patients with small melanocytic choroidal tumors measuring 3 mm in thickness or less. Clinical parameters of the patient and tumor were extracted and analyzed for their relationship to eventual tumor growth and metastasis using a Cox proportional hazards regression model. RESULTS: Tumor growth was documented in 18% of patients. The factors predictive of tumor growth (multivariate analysis) included greater tumor thickness, posterior tumor margin touching optic disc, symptoms of flashes, floaters, and blurred vision, orange pigment on the tumor surface, and the presence of subretinal fluid. The relative risk (rr) was greatest for initial tumor thickness > 2.0 mm (rr 5.2) and posterior margin touching the optic disc (rr 2.6). After adjusting for significant tumor variables, the effect of interventional tumor treatment showed a decreasing risk for tumor growth as compared to continued observation without treatment. Of 1329 patients, 35 (3%) developed metastases. The factors predictive of metastases (multivariate analysis) included: posterior tumor margin touching the optic disc, documented growth, and greater tumor thickness. The relative risk for metastases was greatest for tumor thickness 1.1 3.0 mm (rr 8.8) and growth (rr 3.2). CONCLUSION: Of small choroidal melanocytic tumors measuring 3 mm or less in thickness at the time of initial examination, 18% demonstrate growth and 3% metastasize during the period of followup. Based on this analysis, the clinical features of these tumors can be used to estimate the risk for tumor growth and metastases and assist the clinician with patient management. PMID- 8719684 TI - Histopathologic observations on human eyes following neodymium: YAG laser cyclophotocoagulation for glaucoma. AB - PURPOSE: Although Nd:YAG laser cyclophotocoagulation has been extensively used for nearly a decade in treatment of severe glaucoma, there have been remarkably few reports (each of them extremely brief) of histopathological examination of glancomatous human eyes that had been so treated. We undertook this study to provide further details regarding the nature of the lesions produced in this type of ciliary ablation. METHODS: We chose three representative cases in which a glaucomatous human eye had been subjected to Nd: YAG cyclophotocoagulation, and was subsequently enucleated. To better understand the temporal evolution of the lesions, we selected eyes that were enucleated 1 day, 20 days, and 3 months, respectively, after they had been treated with noncontact Nd: YAG laser cyclophotocoagulation. RESULTS AND CONCLUSIONS: (1) energy levels ranging from 4.4 Joules to 5.6 Joules were effective in producing appropriate lesions; (2) direction of the laser beam 1 to 1.5 mm behind the limbus caused severe destructive lesions of the pars plicata: (3) toward the periphery of the individual treatment sites, the stroma and ciliary muscle continued to exhibit severe degeneration, as did the epithelium lining the valleys between the crests of the ciliary processes; but in those peripheral zones of individual treatment sites, the epithelium lining the crests of the ciliary processes survived and appears normal; (4) bleb-like separations of the ciliary epithelium from the adjacent stroma, particularly along the posterior aspect of the ciliary body lesions, are a prominent early feature of Nd:YAG cyclophotocoagulation; (5) the pigmented epithelium is more vulnerable to laser energy than is the nonpigmented epithelium of the ciliary body; (6) the destruction of the ciliary epithelium is permanent; (7) deeply pigmented persons have more melanocytes in the ciliary body muscle and stroma than do more lightly pigmented individuals, a circumstance that renders the tissues more vulnerable to laser energy; (8) the ciliary muscle was always severely damaged; (9) no scleral injury was observed other than evanescent, focal areas of edema of the deep sclera; and (10) except in the episclera, inflammatory cells were strikingly few in number, a circumstance providing support for the clinical observation that eyes treated with laser cyclophotocoagulation exhibit less of an inflammatory response than do those treated with cyclocryotherapy. PMID- 8719685 TI - Comparison of the configuration of the human anterior chamber angle, as determined by the Spaeth gonioscopic grading system and ultrasound biomicroscopy. AB - PURPOSE: The primary purpose of this investigation was to validate anatomically, if possible, the Spaeth gonioscopic grading system. METHODS: The gonioscopic appearance of the anterior chamber angle of 22 patients was described, using a Zeiss 4-mirror gonioscopic lens; the angle was graded according to the Spaeth gonioscopic grading system (SGGS) previously described elsewhere. This system provides a more comprehensive characterization of the angle than other methods, including the site of iris insertion, the angular approach to the recess and the curvature of the peripheral iris. The same patients were then evaluated with biomicroscopic ultrasound examination of the anterior segment; the nature of the anterior chamber angle was described independently by two observers. The two methods of characterizing the anterior chamber angle were then compared. RESULTS: There was a high correlation between the two different methods of characterizing the anterior chamber angle. Ultrasound biomicroscopy had limitations relating to inability to distinguish between iris-cornea apposition and iris-cornea adhesion, and to exact location of the posterior trabecular meshwork. The angularity of the approach to the anterior chamber angle using the SGGS tended to be underestimated by about 5 degrees. Other aspects showed high correlation. CONCLUSION: The SGGS appears to be an accurate method of characterizing the anatomic appearance of the anterior chamber angle in patients. It also provides a more comprehensive description of the angle that other grading systems. PMID- 8719686 TI - Surgery for upper eyelid retraction, three techniques. AB - BACKGROUND: This paper is based on the author's experience with 145 operations on 181 upper eyelids in 100 patients using three techniques for recession of the upper eyelid. The results include 15 years' experience with adjustable sutures in upper eyelid recession surgery. METHODS: Each chart was reviewed by the author. Patient data was entered into a database and analyzed. The operations used were (1) The prewhitnall levator recession (Trans Am Ophthalmol Soc. 1988: 86: 725 793). This operation is unique in that it is performed on the levator muscle proximal to Whitnall's ligament. (2) Recession of the levator aponeurosis and (3) the Henderson operation (Arch Ophthalmol 1965; 74: 205-216). Adjustable sutures were used in all prewhitnall recessions, about half of the levator aponeurosis recessions and none of the Henderson operations. RESULTS: Complications were divided between over and undercorrection. There were 24% reoperations in 73 patients with thyroid eye disease and 14% reoperations in 17 patients with overcorrected ptosis. The overall reoperation rate was 23%. Adjustable sutures were effective in prewhitnall recessions but less so in levator aponeurosis recessions. There were fewer Henderson operations but the results were good. CONCLUSIONS: The prewhitnall operation is good for marked bilateral eyelid retraction in thyroid eye disease. It is technically challenging but has advantages. Recession of the levator aponeurosis can be used for thyroid eye disease and overcorrected ptosis. The Henderson operation is useful in lesser degrees of eyelid retraction. The need for reoperation is an acceptable occurrence and can be expected in up to one fourth of patients. PMID- 8719683 TI - Diagnostic pars plana vitrectomy report of a 21-year retrospective study. AB - PURPOSE: To review the experience of diagnostic pars plana vitrectomies (PPV). METHODS: The authors reviewed 405 consecutive diagnostic PPV's performed between November 1973 and October 1994. RESULTS: Diagnostic vitrectomy was performed in 215 (53%) of 405 eyes for suspected endophthalmitis. Of those 215 cases, acute inflammation was confirmed in 62 (28.8%), 60 (27.9%) had microbial organisms present and 36 (16.7%) were culture-positive. Microbial organisms were observed microscopically in 31 (20%) of 156 patients suspected of postoperative endophthalmitis. Of those 31 cases, 23 (74%) were gram-positive, eleven (37%) of 30 eyes had organisms associated with glaucoma filtering procedures and 20 (16%) of 126 eyes had organisms with non-filtering procedures. The pooled percentage of eyes that developed postoperative endophthalmitis as a complication during the period July 1990 thru June 1994 is 5 (0.046%) out of a heterogeneous group of 10,898 cases operated on at the Wilmer Eye Institute for cataract, glaucoma, corneal transplant, pars plana vitrectomy and retinal detachment. Bacteria were identified microscopically in 6 (18%) of 34 post-traumatic cases. Microbial organisms were identified in 23 (92%) of 25 cases with an endogenous infection. Patients with endogenous infections had the most fungal infections, and the majority were in males. Neoplasms were diagnosed in 58 (14%) of the 405 cases. The most common neoplasm was ocular lymphoma 42 (72%), 69% of which were in females. Only 42 (48.3%) of 87 patients clinically suspected of having ocular lymphoma, actually had ocular lymphoma. Those negative for lymphoma were significantly older (67.4 +/- 10 years) compared to those with lymphoma (60.4 +/- 14 years) (P = 0.01). CONCLUSION: Diagnostic PPV has proved to be valuable in confirming and establishing various clinical diagnoses. PMID- 8719687 TI - The differential diagnosis and classification of eyelid retraction. AB - PURPOSE: Classification schemes are useful in the formulation of differential diagnoses. Thoughtful commentary has been devoted to the classification of blepharoptosis, but the causes of eyelid retraction have received less attention in published reports. Although eyelid retraction most frequently is associated with Graves' ophthalmopathy, numerous other entities may cause the sign. This study was undertaken to provide a more comprehensive differential diagnosis and classification of eyelid retraction. METHODS: A series of patients with eyelid retraction was studied, and pertinent published reports were reviewed. RESULTS: Forty-four patients with different causes for eyelid retraction are described. Normal thyroid function and regulation were confirmed in all patients in whom Graves' ophthalmopathy could not be excluded by clinical, biochemical, or historical criteria. CONCLUSION: Based on a series of patients and reported cases, a differential diagnosis for eyelid retraction is proposed using a classification system comprising three categories (neurogenic, myogenic, and mechanistic). PMID- 8719688 TI - Retinal detachment following cataract surgery with capsulorhexis. AB - PURPOSE: To estimate the incidence of retinal detachment after cataract surgery with capsulorhexis. METHODS: A consecutive series of 2,150 cataract operations were followed for incidence of retinal detachment. A series of 1,000 patients from this group were analyzed for high risk factors: myopia, age, sex, operative complications and capsulotomy. RESULTS: With minimum one year follow up in 90% of patients the incidence of retinal detachment was 0.25% (5 cases). CONCLUSION: The true incidence of retinal detachment after cataract surgery remains elusive. There is probably a trend toward lower incidence compared to previous reports. PMID- 8719689 TI - Pediatric aphakic glaucoma: a study of 65 patients. AB - PURPOSE: The anterior segment findings of children with glaucoma following pediatric cataract extraction were studied to determine the mechanism of this complication. METHODS: The clinical records of 65 children with glaucoma cared for by the author were studied. Patients with bilateral lensectomy followed by bilateral glaucoma, bilateral lensectomy followed by unilateral glaucoma, and unilateral lens surgery followed by glaucoma were identified and studied. RESULTS: It was determined that vitreous cutting instruments were used for 80% of the lensectomy procedures, with 77% of the lens procedures done in the first year. Eighty-seven percent of the patients were recognized to have glaucoma two or more years after their lensectomy. Preoperative gonioscopy revealed no consistent angle defect, while postoperative gonioscopy revealed a near constant (96%) but variable angle defect characterized by blockage of the trabecular meshwork by an acquired repositioning of the iris against the posterior trabecular meshwork, associated with abnormal pigmentation and synechia formation. CONCLUSION: The results reported document the development of a characteristic acquired filtration angle deformity in children with glaucoma following early lensectomy frequently associated with significant residual lens tissue and unassociated with evidence of chronic anterior segment inflammation. PMID- 8719690 TI - Topical apraclonidine hydrochloride in eyes with poorly controlled glaucoma. The Apraclonidine Maximum Tolerated Medical Therapy Study Group. AB - OBJECT: We determined whether the addition of topical apraclonidine hydrochloride to eyes receiving maximal medical therapy, with inadequate intraocular pressure (IOP) control, and scheduled to undergo surgery, could adequately lower IOP, postponing the need for surgical intervention. DESIGN: A prospective 90 day, multi-centered, placebo-controlled, doublemasked parallel study. PATIENTS: We enrolled 174 glaucoma patients with inadequate IOP control on maximally tolerated medical therapy. All were candidates for either laser trabeculoplasty or invasive surgical intervention. We enrolled only one eye per patient. INTERVENTIONS: We continued to administer maximum-tolerated medical therapy for glaucoma. Patients took the study medication every eight hours. Study medications were either apraclonidine hydrochloride 0.5% or placebo (apraclonidine's vehicle). MAJOR OUTCOME MEASURES: We evaluated IOP, IOP change from baseline, and the number of eyes requiring surgery after the addition of study medication. RESULTS: Sixty one percent of patients treated with apraclonidine maintained adequate IOP control throughout the study, avoiding additional surgery compared to 33.9% patients treated with placebo (P < .001). Apraclonidine treatment resulted in significantly more patients achieving either an additional > or = 20% reduction in IOP from baseline (resulting in an IOP < or = 20 mm Hg) (P < 0.05). The most common ocular complications were conjunctival hyperemia (12.6%), itching and foreign body sensation (6.8%), and tearing (4.5%). The most frequent non-ocular adverse events related to apraclonidine were dry mouth (4.5%) and unusual taste perception (2.2%). CONCLUSIONS: Apraclonidine appears safe and efficacious. It significantly lowered IOP when used in combination with a patient's maximum tolerated medical therapy. This delayed or prevented further glaucoma surgery for at least 90 days in approximately 60% of treated patients. PMID- 8719691 TI - Neurofibromatosis type 1 in children. AB - OBJECTIVE: To document ophthalmic and general characteristics of Neurofibromatosis Type 1 (NF1) in children; and to characterize the iris changes of NF1 including their variability and reliability. DESIGN AND PATIENTS: One hundred and ninety-six patients with NF1 were evaluated for general characteristics; 156 patients underwent prospective eye evaluations; and 151 NF1 patients and controls had iris photography in accordance with a protocol. Masked evaluation of photographs compared to a known diagnosis, and interobserver reliability tests were performed. MAIN OUTCOME MEASURES: Incidence of iris changes consistent with NF1 (Lisch nodules, plus) by age group; spectrum of iris changes found; correlation of iris findings to known diagnosis and Kappa coefficients for interobserver reliability. RESULTS: Iris changes were common in children over age 5, and increased with age; masked evaluation of photographs compared to a known diagnosis yielded fair to poor correlation (Kappa = -.02 to .50); interobserver reliability was poor (Kappa = -.02 to .24, overall .174); iris changes found in NF1 are more diverse than classic descriptions of Lisch nodules, and including a broader spectrum probably increases the sensitivity and decreases the specificity of their diagnostic value. CONCLUSION: Iris changes as a diagnostic marker for NF1 may need rethinking; this study calls to question their reliability and validity. PMID- 8719692 TI - Familial exudative vitreoretinopathy. AB - PURPOSE: To evaluate the natural history of Familial Exudative Vitreoretinopathy (FEVR) with emphasis on the effect of the age of onset on its severity and on the development of late complications such as cataract and retinal detachment. Also, to evaluate affected patients for DNA abnormalities. METHODS: The records of thirty-nine patients with FEVR were studied. All were asked to come in for a final follow-up examination. The referring physician was asked to provide the latest findings for those who could not. On 10 patients, karyotypes were prepared. RESULTS: Only 2 of 28 patients whose onset of symptoms was prior to their third birthday had a final visual acuity of 20/200 or better. Older patients had a better prognosis, because they were more likely to have asymmetrical retinal deterioration with only one eye deteriorating. Preservation of good visual acuity into the teens and later was no guarantee that deterioration would not occur. In 3 eyes of 4 patients who were asymptomatic until 15 years of age, the final visual acuity was counting fingers or worse. In 5 patients, retinal detachment developed 6 to 17 years after apparent stabilization. The karyotype of 10 patients showed no evidence for rearrangement, altered size, translocations or deletions of chromosome 11 or any other chromosome. Of the 31 eyes in patients older than 15 years, 10 (32%) had a significant cataract. Three eyes underwent cataract surgery. CONCLUSIONS: The prognosis for infants with FEVR is extremely poor and the long-term prognosis for patients with a later onset of the condition is guarded. Retinal detachment, macular dragging, and cataract are common late complications which can develop even in patients whose eye findings appear to be stable. Karyotype studies ruled out involvement of chromosome 11 or others at a gross level, but did not exclude them at the location for some genetic defect related to FEVR because single base changes and small deletions or insertions may be undetectable by the methods utilized. PMID- 8719693 TI - Vision development in the monocular individual: implications for the mechanisms of normal binocular vision development and the treatment of infantile esotropia. AB - PURPOSE: The purpose of this research is to study the vision development in monocular individuals so as to better understand normal binocular vision development and to refine the treatment of infants with infantile esotropia. METHODS: Thirty-six subjects with one clinically normal eye and one eye with no vision (no light perception or history of enucleation) are studied. In addition to measurement of standard parameters of development such as visual acuity, measurement of motion processing is made by both optokinetic and electrophysiologic techniques. A comparison is made of vision development among three populations: the monocular population, the normal population, and patients with a history of infantile esotropia. Such comparison is made to study the relative effects of interruption of binocularity and binocular competition. The monocular population represents individuals who have interruption of binocularity, whereas the infantile esotropia population has both interruption of binocularity and binocular competition. RESULTS: The OKN data suggest that the monucular population is more similar to the normal population than the esotropia population. The electrophysiologic data shows a statistically significant difference in the three populations. Motion processing is more fully developed in the monocular population than in the infantile esotropia population when compared to the normal population. CONCLUSIONS: 1. The development of motion processing appears to be particularly vulnerable to abnormal experience during the first year of life. 2. Monocular subjects have a less abnormal motion processing system when compared to patients with infantile esotropia even when monocularity is congenital. 3. The results indirectly support the premise that prealignment alternate occlusion is of benefit to the patient with infantile esotropia prior to realignment. 4. Development of the motion processing system does not necessarily parallel the development of other binocular functions. PMID- 8719694 TI - Histamine, ZO-1 and increased blood-retinal barrier permeability in diabetic retinopathy. AB - PURPOSES: First, to develop an improved retinal capillary endothelial cell culture system which exhibits some of the physiologic features of the bloodretinal barrier; second, to use this model to determine how histamine and chemical conditions of diabetes effect expression of the tight junction protein, ZO-1; and third, to discuss application of the Henle-Koch postulates to the problem of diabetic retinopathy. METHODS: Bovine retinal capillary endothelial cells were exposed to varying serum and growth factor concentrations, as well as astrocyte-conditioned medium, in order to establish a model of the blood-retinal barrier. Cells were also exposed to varying concentrations of histamine, and of insulin and glucose. The expression of ZO-1 tight junction protein was determined by immunocytochemistry and immunoblotting. RESULTS: Modified concentrations of growth factors reduced endothelial cell proliferation, without reducing viability. Astrocyte conditioned medium increased ZO-1 protein content. Histamine reduced ZO-1 protein content. Both high glucose (20mM) and low insulin (10(-12)M) reduced ZO-1 protein content compared to control conditions (5mM glucose and 10( 9) M insulin). CONCLUSIONS: Control of culture conditions results in a more physiologic in vitro model of the blood-retinal barrier. Soluble factors from astrocytes promote tight junction formation. Both histamine and chemical conditions of diabetes diminish tight junction formation. These factors may mediate blood-retinal barrier breakdown in diabetic retinopathy. Henle-Koch postulates for diabetic retinopathy are presented. PMID- 8719696 TI - The dyschromatopsia of optic neuritis: a descriptive analysis of data from the optic neuritis treatment trial. AB - PURPOSE: We sought to characterize the dyschromatopsia of optic neuritis, to determine the type and severity of color defect present and its relation to central vision and spatial acuity, to examine changes in this dyschromatopsia over time, and to determine the applicability of Kollner's rule to patients with optic neuritis. METHODS: We analyzed the raw data on color vision performance as assembled within the Optic Neuritis Treatment Trial (ONTT). The ONTT was designed to evaluate corticosteroids as a treatment for acute demyelinating optic neuritis and to allow long-term outcome and natural history analyses. Between July 1, 1988 and June 30, 1991, 488 patients were enrolled in this trial. All patients underwent extensive neurologic and ophthalmologic examinations including standardized testing of visual function that included testing of color vision. The ONTT population thus afforded a unique opportunity to characterize acquired dyschromatopsias in a large, homogenous, well-characterized cohort of patients with optic neuritis. We used quantitative analysis of FM-100 scores from this patient cohort to determine the severity of the dyschromatopsia, the selectivity of the dyschromatopsia (polarity of errors) and the type of dyschromatopsia (axis of confusion) by employing quadrant analysis of FM-100 scores. RESULTS: The results of high-and low-selectivity analyses of the FM-100 data showed that during the acute phase of optic neuritis, blue/yellow, red/ green, and non selective color defects occurred; among patients with pure defects, blue/yellow defects were more frequent than red/green defects. At 6 months after the acute event, however, analyses showed that red/green defects were more common than blue/yellow defects. Among patients with selective color defects both acutely and at 6 months, the defect was as likely to change over time as remain the same. The likelihood of persistent dyschromatopsia at 6 months was related to the severity of initial central acuity loss, but the type of dyschromatopsia present (red/green versus blue/yellow) was not. CONCLUSIONS: Our results suggest that at the time of the acute attack of optic neuritis, the majority of selective color defects were blue/yellow defects, whereas at 6 months, more of the selective defects were red/green defects, though both types of defects (as well as nonselective defects) were seen acutely and at 6 months. Despite the rigorous inclusion criteria of the ONTT, the large number of patients we studied, correlation of color vision with visual acuity, and longitudinal follow up, this study showed that no single type of color defect was consistently associated with optic neuritis. Demyelinating optic neuritis does not obey Kollner's rule. Moreover, the type of defect present changed in some patients over the course of recovery. Thus, the type of defect may not even be consistent in individual patients as they recover. The type of defect appeared to be related to spatial vision at the time of the test, but the type of defect present at 6 months was not related to the severity of the initial visual loss. Therefore, in evaluating color defects associated with optic neuritis, the level of central visual function must be considered. PMID- 8719695 TI - Ocular manifestations of HIV infection. AB - OBJECTIVE: To evaluate the frequency of ocular complications and the clinical outcomes of these complications in patients with various stages of HIV infection. METHODS: Retrospective review of all HIV-infected patients seen in an AIDS ophthalmology clinic from November 1983 through December 31, 1992. RESULTS: Eleven-hundred sixty-three patients were seen for ophthalmologic evaluation. Of these, 781 had the acquired immune deficiency syndrome (AIDS), 226 had symptomatic HIV infection (AIDs-related complex [ARC]), and 156 had asymptomatic HIV infection. Non-infectious HIV retinopathy was the most common ocular complication, affecting 50% of the patients with AIDS, 34% of the patients with ARC, and 3% of the patients with asymptomatic HIV infection. Cytomegalovirus (CMV) retinitis was the most common opportunistic ocular infection, affecting 37% of the patients with AIDS. Other opportunistic ocular infections, including ocular toxoplasmosis, varicella zoster virus retinitis, and Pneumocystis choroidopathy were all much less common, each occurring in < or = 1% of the patients with AIDS. Treatment of CMV retinitis with either foscarnet or ganciclovir was successful in initially controlling the retinitis. However, relapse represented a significant problem and required frequent re-inductions. As a consequence of the retinal damage associated with relapse, loss of visual acuity occurred. The median time to a visual acuity of 20/200 or worse for all eyes with CMV retinitis was 13.4 months, and the median time to a visual acuity of 20/200 or worse in the better eye was 21.1 months. At last follow-up, 75% of the patients had a final visual acuity of 20/40 or better in at least one eye. Retinal detachments were a frequent ophthalmologic complication of CMV retinitis with a cumulative probability of a retinal detachment in at least one eye of 57% at 12 months after the diagnosis of CMV retinitis. Herpes zoster ophthalmicus developed in 3% of the overall series and was seen in all stages of HIV infection. Fifty-six percent of the cases of ocular toxoplasmosis had simultaneous toxoplasmic cerebritis. Ocular toxoplasmosis responded to standard anti-microbial therapy. Varicella zoster virus retinitis, when manifested by the acute retinal necrosis (ARN) syndrome, responded to intravenous acyclovir therapy. Conversely, in a limited number of patients with the progressive outer retinal necrosis syndrome, the disease responded poorly to intravenous acyclovir therapy, but appeared to respond to combination foscarnet and acyclovir therapy. Neuro-ophthalmic lesions were present in 6% of the patients with AIDS. The most common cause of a neuro-ophthalmic lesion was cryptococcal meningitis, and 25% of the patients with cryptococcal meningitis developed a neuro-ophthalmic complication. CONCLUSIONS: Ocular manifestations are common in patients with AIDS. CMV retinitis represented a major vision-threatening problem in these patients. While available therapy was successful in initially controlling the retinitis, the phenomenon of relapse resulted in some degree of long-term visual loss. Preservation of the patient's visual acuity in at least one eye was generally successful. Other opportunistic ocular infections were substantially less common than CMV retinitis but require aggressive therapy. PMID- 8719699 TI - Functional and molecular analysis of glutamate-gated channels by patch-clamp and RT-PCR at the single cell level. AB - In the central nervous system (CNS) rapid excitatory neurotransmission is mainly mediated by ligand gated, cationic channels activated by glutamate. Three main subtypes of glutamate-gated channels have been characterized by pharmacological studies. They have been named according to their preferred agonist, N-methyl-D aspartate (NMDA), high affinity kainate and alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionate (AMPA). Furthermore, a large diversity within each class of glutamate-gated channels has been revealed by the molecular cloning of multiple subunits and their spliced and edited variants (for review see Wisden and Seeburg, 1993). These subunits can potentially form different oligomeric complexes with diverging properties. A crucial question is therefore to determine the actual subunit composition of naturally occurring glutamate receptors. We have combined patch-clamp recording, reverse transcription (RT) and PCR to correlate, at the single cell level, the pattern of subunits expression with the functional properties of native glutamate receptors. We describe here results obtained on the AMPA receptors of hippocampal neurones and on the NMDA receptors of cerebellar granule cells which show that the subunit composition of these two types of receptors explains some of their functional properties. Furthermore, our data also indicate that the expression of NMDA receptor subunits during the postnatal development of cerebellar granule cells is regulated by an activity dependent mechanism. PMID- 8719697 TI - Simultaneous evaluation of tear turnover and corneal epithelial permeability by fluorophotometry in normal subjects and patients with keratoconjunctivitis sicca (KCS). AB - PURPOSE: To simultaneously determine the tear turn over rate (TT) and corneal epithelial permeability (Pdc) in normal subjects and patients with KCS by a single-drop fluorophotometric technique using disodium fluorescein (DSF) or carboxyfluorescein (CF). METHODS: DSF was instilled in one eye chosen at random and CF in the fellow eye of 13 normal subjects and in 13 patients with KCS. TT and Pdc were determined using a single-drop technique on a Fluorotron Master. RESULTS: In normals and KCS subjects, TT was found to be independent of age and sex and essentially identical for DSF and CF. TT was approximately 42% lower in KCS subjects than normals (Table). TT was independent of Schirmer's I and Pdc. [table: see text] Pdc values were similar for DSF and CF in normals and increased with age. In KCS subjects, Pdc was 3-4 times higher compared to normal subjects (p < 0.01) and was directly correlated with the severity of corneal punctate staining. Pdc was independent of TT. CONCLUSION: Patients with KCS are more susceptible to the therapeutic and toxic effects of topical medications and preservatives due to increased corneal epithelial permeability. With the decreased TT in KCS patients, there is also slower elimination of substances from the tear film. This, combined with the increase in epithelial permeability, puts the KCS eye doubly at risk for the toxic effects of topically applied substances. PMID- 8719700 TI - Single cell reverse transcription-polymerase chain reaction (RT-PCR) and the GABA A receptor. PMID- 8719701 TI - Analysis of molecular determinants in native AMPA receptors. PMID- 8719702 TI - Intra- vs extracellular calcium regulation of neurotransmitter-stimulated phosphoinositide breakdown. AB - The dependence on Ca2+ of basal, glutamate- and carbachol-stimulated phosphoinositide (PI) turnover was studied on 8-day old rat brain synaptoneurosomes. For that purpose, intracellular and extracellular Ca2+ concentrations were buffered by bis-(alpha-aminophenoxy)-ethane-N,N,N',N' tetraacetic acid, in its tetra(acetoxymethyl)-ester form (BAPTA-AM) and in its free acid form (BAPTA), respectively. The effects of both forms of the calcium chelator intracellular and extracellular Ca2+ buffering on intracellular and extracellular Ca2+ concentration ([Ca2+]i and [Ca2+]e) were determined with fluorimetric assay using fura2, either in its acetoxymethyl ester form (fura2-AM) or in its free acid form. Intracellular chelation of Ca2+ ions with BAPTA-AM induced a dose-dependent reduction of the [Ca2+]i. Basal inositol phosphate (IP) formation was slightly affected by this [Ca2+]i buffering, while glutamate and carbachol stimulations of PI hydrolysis were similarly diminished. Chelation of extracellular Ca2+ ions with BAPTA produced a reduction of both [Ca2+]e and [Ca2+]i. Basal IP accumulation was maximally inhibited by 50%. The carbachol induced PI hydrolysis was completely inhibited in the presence of 200 microM BAPTA, while a substantial residual glutamate-elicited IP response remained (40% of the control response). It is concluded that [Ca2+]i of synaptoneurosomes is not critical for basal and neurotransmitter-stimulated IP formation, whilst [Ca2+]e is critical. Glutamate may, in part, stimulate PI breakdown in a Ca(2+) insensitive way. PMID- 8719703 TI - Cat proenkephalin-A does not contain the opioid octapeptide. AB - The sequence of a large cDNA fragment of proenkephalin-A from the cat adrenal medulla was obtained using reverse transcription followed by polymerase chain reaction, and cloning. This cDNA encompasses the region normally containing all the opioid peptides, except the C-terminal heptapeptide. As with other species, cat proenkephalin-A contains four conserved copies of (Met5)-enkephalin, and one of (Leu5)-enkephalin, flanked by processing sites of paired basic amino acids. However, significant differences were found in the nucleotide and deduced amino acid sequences in the region of the octapeptide. In particular, the essential tyrosyl residue is substituted by a histidyl residue, making it unlikely that the cat equivalent would have opioid activity. Furthermore, the peptide is not flanked by paired basic residues, suggesting it is not processed. PMID- 8719698 TI - Progression of myopia. AB - BACKGROUND: Myopia is an important public health problem because it is common and is associated with increased risk for chorioretinal degeneration, retinal detachment, and other vision-threatening abnormalities. In animals, ocular elongation and myopia progression can be lessened with atropine treatment. This study provides information about progression of myopia and atropine therapy for myopia in humans. METHODS: A total of 214 residents of Olmsted County, Minnesota (118 girls and 96 boys; median age, 11 years; range, 6 to 15 years) received atropine for myopia from 1967 through 1974. Control subjects were matched by age, sex, refractive error, and date of baseline examination to 194 of those receiving atropine. Duration of treatment with atropine ranged from 18 weeks to 11.5 years (median 3.5 years). RESULTS: Median follow-up from initial to last refraction in the atropine group (11.7 years) was similar to that in the control group (12.4 years). Photophobia and blurred vision were frequently reported, but no serious adverse effects were associated with atropine therapy. Mean myopia progression during atropine treatment adjusted for age and refractive error (0.05 diopters per year) was significantly less than that among control subjects (0.36 diopters per year) (P < .001). Final refractions standardized to the age of 20 years showed a greater mean level of myopia in the control group (3.78 diopters) than in the atropine group (2.79 diopters) (P < .001). CONCLUSIONS: The data support the view that atropine therapy is associated with decreased progression of myopia and that beneficial effects remain after treatment has been discontinued. PMID- 8719704 TI - Long-term neuroleptic treatments counteract dopamine D2 agonist inhibition of adenylate cyclase but do not affect pertussis toxin ADP-ribosylation in the rat brain. AB - We have investigated the response of adenylate cyclase to GTP and to dopamine (DA) in striatal membranes of rats treated for 3 weeks with chlorpromazine or haloperidol, and further measured the level of Gi (an inhibitory GTP-binding protein) or Go (a similar GTP-binding protein of unknown function) in 3 areas (cerebral cortex, striatum and hippocampus) utilizing pertussis toxin-catalyzed ADP ribosylation. In saline-treated control membranes, GTP exerted a biphasic effect on basal and DA-stimulated enzyme activity--peak levels of stimulation by DA plus GTP were observed at 1 microM GTP. Conversely, dopaminergic inhibitory effects at 10-100 microM GTP were completely attenuated in chlorpromazine or haloperidol-treated membranes. D2 inhibition of adenylate cyclase by the selective D2 agonist PPHT was also attenuated due to these neuroleptic treatments, while an increase in D2 receptor binding was observed. The pertussis toxin ADP-ribosylation of G-proteins (Gi/Go) did not differ significantly in any area. This indicates that long-term neuroleptic treatments increased D2 receptor binding, but attenuated D2 inhibition of adenylate cyclase, and exercised no influence on pertussis toxin ADP-ribosylation. PMID- 8719705 TI - Solubilization of [35S]lanthionine ketimine binding sites from bovine brain. AB - Lanthionine ketimine (LK) binding sites were solubilized from bovine brain membranes using 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS) and Triton X-100. 10 mM CHAPS in 0.5 M potassium phosphate, pH 7.0, containing 20% glycerol was selected to solubilize LK binding entities. Some properties of CHAPS-solubilized LK binding sites have been studied. The CHAPS-solubilized preparation appeared to contain a homogenous population of binding sites for [35S]LK. Binding properties indicated that the solubilized binding sites were similar to the membrane-bound sites. [35S]LK specific binding was inhibited by other structurally related ketimines obtaining a similar rank order of inhibition for the soluble and the membrane-bound preparations. The successful solubilization of [35S]LK binding sites is a useful starting point for the purification of this binding protein. PMID- 8719706 TI - Autophosphorylation as a possible mechanism of calcium/calmodulin-dependent protein kinase II inhibition during ischemia. AB - Cardiac arrest induced rat brain ischemia of 15 min duration produces a rapid and profound decrease in activity of calcium/calmodulin stimulated protein kinase (CaM-KII). In contrast to that, the total amount of enzyme protein remains stable as revealed by Western blotting (alpha subunit specific) analysis. Ischemic insult also results in translocation of the enzyme toward plasmatic membranes, reducing its content in soluble (cytosolic) fraction down to 7% with respect to 50% of control. The qualitatively similar translocation can be achieved by autophosphorylation of the control enzyme in vitro. Moreover, severely reduced response of immunoprecipitated enzyme to autophosphorylation observed after ischemia ex vivo probably reflects the higher level of its endogenous phosphorylation during the insult. The results strongly suggest that among various possible mechanisms of postischemic CaM-KII inhibition the most probable would be that involving abnormal or irreversible phosphorylation of the enzyme molecule. It would consequently block or inhibit the autophosphorylation/dephosphorylation cycle of endogenous CaM-KII interconversion necessary for its full catalytic activity. PMID- 8719707 TI - Age-related changes of the ganglioside long-chain base composition in rat cerebellum. AB - The ganglioside mixture from the cerebellum of young, 6 month old and two years old rats, was fractionated by reversed phase high performance liquid chromatography, each ganglioside homogeneous in the oligosaccharide chain as well as in the long-chain base being subsequently quantified. Two long-chain bases, LCB, were components of the five major gangliosides GM1, GD1a, GD1b, GT1b and GQ1b, these being the C18:1 LCB and C20:1 LCB. The content of C20:1 ganglioside molecular species was lower than that of the C18:1 one. In very young animals, day 8, the C20:1 ganglioside species represented about 8% of the total ganglioside content, then they progressively increased and reached, at 2 years, about 42% of the total. C18:1 GD1a and C18:1 GT1b, were the major species in young animals and reached their highest content at day 29, being 1.45 and 1.28 nmol/mg protein, respectively. The content of these two species decreased in adult and old animals and at two years it was 0.71 and 0.82 nmol/mg protein, respectively. PMID- 8719708 TI - NMDA receptor antagonist MK-801 down-regulates rat striatal proenkephalin and protachykinin mRNAs. AB - Using quantitative in situ hybridization, a significant decrease in expression of proenkephalin (27.1%) and of protachykinin mRNAs (20.0%) is observed in the rat caudate-putamen 14 days after daily intraperitoneal administration of N-methyl-D aspartate receptor antagonist MK-801, 2 mg/kg. PMID- 8719709 TI - The novel sigma ligand JO 1994 protects against ischaemia-induced behavioural changes, cell death and receptor dysfunction in the gerbil. AB - To assess the effects of the novel sigma ligand JO 1994 on behavioural, histological and autoradiographical changes following global ischaemia, the Mongolian gerbil was used. Three experiments were carried out and in each case ischaemia was induced by bilateral carotid occlusion (BCO) for 5 min. In the first experiment we examined the effects of JO 1994 administered at doses of 0.25, 0.5 and 1 mg/kg i.p. 1 h before 5 min BCO on histological parameters 96 h after surgery. In the second experiment the effects of JO 1994 administered at doses of 2.5, 5, 10 and 20 mg/kg i.p. 1 h before 5 min BCO on locomotor activity 24, 48 and 72 h after surgery and on histological parameters 96 h after surgery was examined. In the third experiment the effects of JO 1994 (2.5 and 5 mg/kg i.p.), BMY 14802 (1 and 10 mg/kg i.p.) and MK-801 (2.5 mg/kg i.p.) administered 30 min, 6, 24, 48, 72, 96 and 120 h post-surgery on the densities of M1 and M2 muscarinic receptors in 35 brain regions, 7 days after surgery was examined. Results indicated that 5 min bilateral carotid occluded animals were hyperactive 24, 48 and 72 h after surgery. JO 1994 attenuated this hyperactivity. Extensive neuronal death was observed in the CA1 layer of the hippocampus in 5 min BCO animals 96 h after surgery. The low doses of JO 1994 (0.25, 0.5 and 1 mg/kg) had no effect on the ischaemia-induced cell death. However JO 1994 (2.5, 5, 10 and 20 mg/kg i.p.) protected against the neuronal death of cells in the CA1 layer (P < 0.01-0.03). There was a large loss of M1 and M2 receptors in the CA1 regions of the hippocampus. MK-801, BMY 14802 and JO 1994 provided significant (P < 0.01) protection against this ischaemia-induced receptor loss. PMID- 8719710 TI - Changes in proteasome activity following transient ischemia. AB - Succinyl-Leu-Leu-Val-Tyr-4-methylcoumaryl-7-amide (Suc-LLVY-MCA) hydrolyzing activities of the 20S and 26S proteasomes in the gerbil cortex following transient forebrain ischemia were examined. Using extraction solutions without ATP, only 20S proteasome activity was noted after separation with glycerol gradient centrifugation. When these extracts were incubated with ATP and an ATP regenerating system prior to glycerol gradient separation, both 20S and 26S proteasome activities were detected. Following 10 min of ischemia, the activity of the 26S proteasomes decreased, whereas the 20S proteasome activity increased after 30 min of reperfusion. These changes returned to the control level after 1 h. The active 26S proteasomes were formed with ATP-dependent association with the 20S proteasomes and several subunits and the 26S proteasomes degraded ubiquitin protein conjugates. These results indicate that proteasome activity might not be irreversibly impaired after transient ischemia. However, transient inhibition of ATP-dependent conversion of 20S to 26S proteasomes in vitro must be one of the causes of the accumulation of the ubiquitin-protein conjugates in the early reperfusion period. PMID- 8719711 TI - Characterization of [2-3H]deoxy-D-glucose uptake in retina and retinal pigment epithelium of normal and diabetic rats. AB - The outer blood-retinal barrier which results from the tight junctions between retinal pigment epithelial cells (RPE) restricts the flow of nutrients reaching the retina. We characterize the transport of [2-3H]deoxy-D-glucose (2-DG) across isolated mammalian neural retina and RPE in terms of their kinetics constants. In addition, the effect of insulin on glucose transport was studied by using streptozotocin-induced diabetic rats. RPE accumulates 2-DG by a temperature sensitive and energy-dependent complex kinetics mechanism. The retina takes up 2 DG by an energy and Na(+)-dependent saturable system with an apparent Km of 2 mM. Insulin induced an increase of 2-DG uptake by normal retina. The retina of diabetic rats shows lower levels of 2-DG accumulation. These levels can be returned to the normal ones by exposure to insulin. Although insulin does not affect, significantly, 2-DG accumulation by RPE, 2-DG uptake of RPE from diabetic rats shows a normal saturable kinetics with an apparent Km of 20 mM. Those findings suggest the presence of different types of glucose transporters in retina and RPE. Insulin-sensitive glucose transport in retina might be involved in the manifestation of diabetic retinopathy. PMID- 8719712 TI - Changes of GTP binding proteins, not neurofilament-associated proteins, in the brain of the neurofilament-deficient quail, "Quiver". AB - A neurofilament (NF)-deficient mutant of the Japanese quail was named "Quiver", as it showed generalized quivering as a clinical sign. NF consists of three major subunits; low, middle and high. We previously reported that the noradrenaline and 5-hydroxytryptamine content in the neostriatum of the Quiver's brain was different from that in the normal quail, although disappearance of the three NF proteins occurred in all areas of the Quiver's brain. Thus, NF-related proteins may show considerable changes in the specific sites of Quiver's brain. In this study, an examination was made of the changes in NF-related proteins in the Quiver, by immunoblotting analysis. The amounts of cyclin-dependent kinase 5 (cdk5), which phosphorylates NF proteins, and tau which is a substrate of cdk5, in the neostriatum of the Quiver, were essentially the same as those in the normal quail, although NF proteins could not be detected in the Quiver. The amount of alpha-tubulin in the Quiver's brain was similar to that in the normal quail. Next, we investigated the changes of GTP binding (G) proteins in the Quiver's brain, because cytoskeletal components such as tubulin and F-actin bind with G proteins. [32P]ADP-ribosylation of G proteins (Gs by cholera toxin and Gi/G0 by pertussis toxin) in the neostriatum of the Quiver increased significantly, although alpha subunits of G proteins showed no change by immunoblotting analysis. The ratios of the trimer form in G proteins thus appear to increase more in the NF-deficient Quiver brain than in the brain of the normal quail. The G proteins-mediated adenylate cyclase activities were the same in the brain of both the Quiver and the normal quail. Possible interactions between NFs and G proteins are discussed. PMID- 8719713 TI - Amniotic fluid gram stain and leukocyte count in the prediction of intrauterine infection in preterm prelabour amniorrhexis. AB - The purpose of the study was to examine the sensitivity and specificity of the amniotic fluid Gram stain and leukocyte count in the prediction of positive fetal blood and amniotic fluid cultures in 80 patients with preterm prelabour amniorrhexis. Amniocentesis and cordocentesis were performed and amniotic fluid and fetal blood were cultured for aerobic and anaerobic bacteria. Amniotic fluid was also cultured for Ureaplasma urealyticum and Mycoplasma hominis. The sensitivity of the Gram stain in the prediction of positive fetal blood and amniotic fluid cultures was 50 and 40% and the respective false positive rates were 12 and 4%. In the detection of aerobic or anaerobic infection of the amniotic fluid, the sensitivity and false positive rate of the Gram stain were 80 and 3% respectively. This compared favourably with the respective values of 66 and 35% for amniotic fluid leukocyte count > 30/mm3. Positive amniotic fluid Gram stain provides useful prediction of intra-uterine infection with aerobic or anaerobic organisms. PMID- 8719714 TI - Increased amniotic fluid alpha-fetoprotein in patients with fetal nuchal edema. AB - The alpha-fetoprotein concentration in the amniotic fluid of 51 fetuses with isolated nuchal edema and normal karyotypes was compared to that of the next 51 patients of comparable gestational age with normal ultrasounds and karyotypes. The mean alpha-fetoprotein in amniotic fluid was 1.26 +/- 0.45 in the nuchal edema group and 1.01 +/- 0.3 in controls (t = 3.287, p = 0.002). Acetylcholinesterase evaluation was indicated by abnormally elevated alpha fetoprotein in 4% of amniotic fluid samples from the study group and none of the controls. We concluded that alpha-fetoprotein is higher in amniotic fluid from pregnancies with isolated fetal nuchal edema and normal karyotype. Possible leakage of protein from enlarged cervical lymphatics associated with fetal nuchal edema may explain these findings. PMID- 8719715 TI - Comparison of chorion villus sampling and early amniocentesis for karyotyping in 1,492 singleton pregnancies. AB - This was a prospective study in 1,492 women with singleton viable pregnancies undergoing chorion villus sampling (CVS) or early amniocentesis (EA) at 10-13 weeks of gestation because of advanced maternal age, parental anxiety or family history of chromosomal abnormality in the absence of parental chromosomal rearrangement. They were offered the option of having CVS or EA, or to be randomised into one of the two tests. CVS was performed in 652 cases (375 by choice and 277 by randomisation) and EA was performed in 840 cases (562 by choice and 278 by randomisation). Both procedures were carried out by transabdominal ultrasound-guided insertion of a needle (20 gauge) using a free-hand technique. The two techniques were similar in providing a sample (CVS 99.3%, EA 100%) and in giving a non-mosaic cytogenetic result (CVS 97.5%, EA 97.9%). Spontaneous loss (intrauterine or neonatal death) after EA (total group 4.9%, randomised subgroup 5.8%) was significantly higher than after CVS (total group 2.1%, randomised subgroup 1.8%; difference 2.8%, 95% CI 1.3-4.3%, and difference 4%, 95% CI 1.3 6.7%). The gestation at delivery and birth weight of the infants after EA and CVS were similar. In the EA group the incidence of talipes equinovarus (1.66%) was higher than in the CVS group (0.48%), but this difference was not significant. PMID- 8719716 TI - Multiple marker screening in multifetal gestations: failure to predict adverse pregnancy outcomes. AB - The objective of the study was to assess whether the association known in singleton pregnancies of high maternal serum alpha-fetoprotein (AFP) or human chorionic gonadotropin (HCG) with adverse outcomes applies also to multifetal gestations. Maternal serum AFP and HCG were evaluated in 207 multifetal pregnancies. High values were defined as a maternal serum AFP of > 4.5 and a HCG of > 4.0 multiples of the median (MoM), with appropriate adjustments. Results were correlated with premature delivery, stillbirths, or pregnancy termination for fetal anomalies. There were 10 stillbirths, 7 terminations of pregnancy for fetal anomalies, and 66 premature deliveries in the study group. Maternal serum AFP was somewhat higher in abnormal pregnancies than in those with normal outcome (3.4 vs. 2.5 MoM, respectively, NS). A high AFP level was found in 6% of pregnancies with adverse outcomes and in 4% of uncomplicated gestations (NS). High HCG values, also observed in 5% of cases, were all associated with normal outcome. Multiple marker screening suggested an increased risk for aneuploidy in 9% of patients, all of whom were euploid on amniocentesis karyotypes. Maternal serum screening in multiple gestations is confounded by the differing contributions of fetuses, and abnormal results cannot reliably predict adverse pregnancy outcomes. PMID- 8719717 TI - Women's informed choice of prenatal diagnosis: early ultrasound examination routine ultrasound examination-age-independent amniocentesis. AB - The antenatal clinics in the catchment area of the Department of Obstetrics and Gynecology, University Hospital, Lund, were divided into a study group and a control group. At the study antenatal clinics during a 15-month period 1,004 pregnant women received a written and verbal information that a routine ultrasound examination during gestational week 18 should be considered as a prenatal diagnostic method. They even got information on alternatives: to decline all, to have an early abdominal ultrasound examination, or to have both a routine ultrasound examination and an amniocentesis performed. At the control antenatal clinics, 1,408 pregnant women received standard information. No women in either group chose not to have an ultrasound examination performed, but 1% chose an early abdominal examination in the study group. The percentage of women older than 35 years who asked for an amniocentesis was equal in the two groups. However, women younger than 35 years in the study group requested amniocentesis significantly more frequently than the women in the control group. PMID- 8719718 TI - Single-needle insertion technique for thoracocentesis for bilateral pleural effusions. AB - A 28-week fetus with hydrops and bilateral pleural effusions underwent drainage of both pleural spaces via a single-needle supradiaphragmatic insertion technique. The traditional approach was not feasible due to a persistently unfavorable fetal position. The dependent pleural effusion was drained by a needle which coursed through the nondependent pleural effusion. The needle traversed the midline pleura through an avascular plane. Thereafter the needle was withdrawn into nondependent pleural effusion which was drained. Both lungs expanded with favorable neonatal outcome. PMID- 8719719 TI - Internalization of pleuroamniotic shunt causing neonatal demise. AB - Internalization of a pleuroamniotic shunt previously has not been associated with significant fetal or neonatal problems. We report a case in whom the shunt internalized and compressed the lung hilum. A review of the complications of antenatal pleuroamniotic shunting is also presented. PMID- 8719720 TI - Fetal pulse oximetry during labor of 62 patients. AB - The objectives of the study were to assess the ability of a fetal pulse oximeter to measure the fetal oxygen saturation (SPO2) continuously during labor and to assess the correlation between readings of fetal pulse oximetry and cord blood gases at birth. The Nellcor N-400 Fetal Oximeter was used in 62 women prospectively. The mean SPO2 was unchanged at the different stages of normal labor. 53.3 +/- 9.1 to 50.1 +/- 11.8% (n = 50). The SPO2 recorded during the pushing phase of stage II (n = 40) was correlated significantly with umbilical cord arterial pH (r = 0.55; p = 0.0002), venous pH (r = 0.63; p = 0.0001), venous PCO2 (r = 0.47; p = 0.002) and venous PO2 (r = 0.39; p = 0.01). SPO2 was also correlated with scalp pH (r = 0.52; p < 0.05; n = 21). No side effects were noted. Fetal pulse oximetry could be useful to assess fetal oxygenation during labor and at birth. PMID- 8719721 TI - Correction of hemodynamic abnormalities by vesicoamniotic shunting in familial congenital megacystis. AB - Vesicoamniotic shunting is typically reserved for treatment of fetal obstructive uropathy. We report a case of congenital megacystis without anatomic urinary obstruction in whom a vesicoamniotic shunt was used to improve fetal hemodynamics as measured by Doppler velocimetry. PMID- 8719722 TI - Transverse cerebellar diameter and transverse cerebellar diameter/ abdominal circumference index for assessing fetal growth. AB - In a prospective study of 330 pregnant women, measurements of the transverse cerebellar diameter (TCD), abdominal circumference (AC), biparietal diameter (BPD), head circumference (HC) and TCD/AC ratio and HC/AC ratio were obtained using conventional ultrasonography between 16th and 41st weeks of gestation. Measurement of BPD yielded a strong correlation with gestational age (r = 0.9880 and R2 = 0.9761) by cubic regression analysis. The measurement of TCD also had a very close relation with gestational age (r = 0.9767 and R2 = 0.9539). The ratio between TCD and AC was calculated and found to be 0.1436 +/- 0.0106 (SD) which remained fairly constant throughout pregnancy, while the BPD/AC ratio declined. The 5th and 95th percentiles for this ratio were 0.1279 and 0.1603. Ten of eleven fetuses with TCD/AC ratios exceeding 2 SD (0.1648) were found to have asymmetrical intrauterine growth retardation upon neonatal examination. It is demonstrated that both BPD and TCD measurements are reliable techniques that correlate well with gestational age, and the TCD/AC ratio is valuable in identifying babies with asymmetrical intrauterine growth retardation. PMID- 8719723 TI - Prenatal diagnosis of hepatic hemangioma. AB - We report the prenatal diagnosis of a hepatic hemangioma presenting as a heterogeneous mass (6 x 5 cm) of the fetal liver, thickly septated with calcifications. Pulsed Doppler confirmed the vascular pattern of the mass. Subsequent postnatal examinations and follow-up have documented the spontaneous regression of this benign tumor. Antenatal diagnosis of fetal liver masses and management after delivery are discussed. PMID- 8719724 TI - The congenital mesoblastic nephroma: a case report of prenatal diagnosis. AB - The congenital mesoblastic nephroma is a very rare benign congenital renal tumor. It is the most common renal tumor before the age of 6 months (50%) and it constitutes only 5% of renal tumors before 15 years. The authors report a case of prenatal diagnosis of congenital mesoblastic nephroma revealed by an acute polyhydramnios at 33 weeks of pregnancy. After a preterm labor, the patient delivered at 35 weeks. The newborn underwent a radical nephrectomy. No recurrence was noticed at 10 months. This case of prenatal diagnosis is compared to the 12 cases previously reported. The prognosis of CMN depends on histologic findings, but also on the severity of prematurity induced by the polyhydramnios. The main treatment of this pathology if diagnosed during pregnancy remains the prevention of preterm labor, and after birth the removal of the kidney. PMID- 8719725 TI - Endoscopic tracheal obstruction with an expanding device in a fetal lamb model: preliminary considerations. AB - Tracheal obstruction to promote lung growth may be a less aggressive alternative to open fetal surgery in the antenatal treatment of congenital diaphragmatic hernia. Herein, we explore the feasibility of placing an occluding device through fetal tracheoscopy. A self-expanding umbrella allowed adequate sealing of the tracheal lumen even as the tracheal diameter more than doubled between 110 and 138 days of gestation (term = 145 days) in a sheep model. Distal intratracheal pressures after umbrella placement, and lung weight at delivery, were comparable to those after formal tracheal ligation. PMID- 8719726 TI - Russell-Silver syndrome: an explanation for discordant growth in monozygotic twins. AB - Severe intrauterine growth retardation and suggestive dysmorphic and malformation syndrome in 1 twin with a normal karyotype during a monozygotic, monochorionic pregnancy led to the diagnosis of discordant Russell-Silver syndrome (RSS). Placental anomalies (velamentous cord insertion, single umbilical artery, placental hypotrophy) for the small-for-date twins and a twin-twin transfusion syndrome confirmed at 21 weeks of amenorrhea suggest that early hemodynamic disorders may reveal or trigger RSS which has been considered to be of genetic origin. The prognosis for RSS is favorable, except for diminished adult size (less than -2 standard deviations). PMID- 8719727 TI - Sirenomelia and situs inversus: case report and review of the literature. AB - We report a case of sirenomelia of the symelia dipus type which is associated with complete situs inversus. Sirenomelia is difficult to diagnose prenatally because of a frequently associated oligohydramnios. Symelia dipus type adds to the difficulty of the diagnosis, since the bones of the thighs and legs are fully formed. In fact, a close examination of the cases of sirenomelia previously reported in the literature shows that a prenatal diagnosis was achieved only when the oligohydramnios was no more than moderate and/or the associated malformation was of apus or unipus type. We propose a systematic amnioinfusion when one suspects symelia unipus type. Finally, the pathogenesis of sirenomelia is discussed, as the presence of a complete situs inversus in the case reported here could illuminate the debate concerning the origin of the malformation. PMID- 8719728 TI - Carcinoembryonic antigen: the old dog finally learns new tricks. PMID- 8719729 TI - Management of recurrent colorectal cancer: another look at carcinoembryonic antigen-detected recurrence. AB - There is currently controversy regarding the use of serum carcinoembryonic antigen (CEA) in the follow-up of colorectal cancer. This article reviews the most recent clinical data regarding CEA and discusses the rationales for its use in the follow-up of colorectal cancer. The patterns of colorectal cancer recurrence and their variable association with CEA elevation are presented. The contributions of the new imaging modalities, including radiolabeled antibodies, are reviewed and information on the prognosis following resection of recurrences is provided. A practical approach to colorectal cancer recurrences is suggested in two algorithms. PMID- 8719730 TI - Laparoscopic interventions in the gut: yesterday, today, and tomorrow. AB - The development of laparoscopic interventional surgery has brought about a revolution in general surgery over the past 5 years. Laparoscopic cholecystectomy has now become the treatment of choice for symptomatic cholelithiasis because of a reduction in access trauma, resulting in less postoperative pain and a faster recovery. Laparoscopic fundoplication for gastroeosophageal reflux also looks to be a promising procedure which will probably become generally accepted. Laparoscopic hernia repair, although widely practised, has raised question marks because of doubt about recurrence rates and major complications such as intestinal obstruction. Laparoscopic colectomy has similarly raised concerns about complication rates and tumour recurrence rates. Laparoscopic appendicectomy offers marginal benefits over open appendicectomy. Virtually every other intra abdominal procedure has been performed laparoscopically, but most require further evaluation. PMID- 8719731 TI - Gastroduodenal dyspepsia: a personal view integrating clinical, endoscopic, histological and management criteria. The spiro syndrome. AB - I am presenting my view on how to approach the difficult subject of dyspepsia, based on my personal experience and the writings and teachings of Howard M. Spiro. Symptoms arising from the esophagus, and called esophageal dyspepsia, are separated from symptoms arising from the stomach, designated as gastroduodenal dyspepsia. The holistic approach to patients with dyspepsia, and designated Spiro syndrome, is the main purpose of this report. I am introducing a newly defined classification, and criteria, using an interchangeable, standardized nomenclature, to be used by the clinician, endoscopist, and pathologist for diagnosing and managing the causes of gastroduodenal dyspepsia. There are five clinical/endoscopic/histological categories to be considered as possible causes of gastroduodenal dyspepsia. Often these entities are found to be asymptomatic, or a combination may cause symptoms: (1) idiopathic dyspepsia--normal endoscopy and histology; (2) congestive gastropathy/duodenopathy; (3) gastritis/duodenitis; (4) peptic ulcer crater, and (5) gastric cancer. I believe this holistic, unifying approach to diagnosis and management of dyspepsia will enhance the communication between physicians and help standardize the terminology for clinical investigation. PMID- 8719732 TI - Dietary fiber: fact and fiction. AB - There have been many conflicting reports about the benefits of dietary fiber. Dietary fiber has been touted to provide remarkable benefits: from the control of functional bowel disease to the prevention of cancer. We have reviewed all prominent studies on the benefits of dietary fiber and examined the nature of their conflicting results. Recommendations for dietary fiber intake are also discussed. PMID- 8719733 TI - Towards understanding the process of intestinal adaptation. AB - When bowel is lost due to disease or surgery, the residual bowel increases its functional capacity in order to compensate for the loss of absorptive capacity. The success and extent of this adaptive process is critical to recovery. This article reviews the current understanding of the intestinal adaptation. Adaptation is a complex process which as yet is poorly understood and thus it is difficult to develop strategies to enhance the out-come of the adaptation phase. Extensive loss of bowel brings about functional as well as morphologic changes in the remaining intestine. The exact relationship between histologic changes and function is still unknown and some studies show that histologic changes may precede functional changes. Both nonnutritive and nutritive factors have been identified as major stimuli to the adaptive process. The importance of oral intake as a positive stimulus has been demonstrated and the role of diet composition rather than diet complexity being recognized. Recent studies attempt to understand the molecular basis of intestinal adaptation by seeking to characterize the nature of humoral factors involved and to establish alterations in the patterns of gene expression. These new approaches may facilitate the identification of both nutritional and pharmacological methods to manipulate the intestinal adaptation process. PMID- 8719734 TI - Influence of insecticidal plant materials used during storage on sensory attributes and instrumental hardness of dry edible beans (Phaseolus vulgaris L.). AB - Three plant products with known insecticidal properties, a dry extract of flowers of Chrysanthemum cinerariaefolium (Trevir.) Vis. produced in Rwanda, an ethanol extract of seeds of neem, Azadirachta indica A. Juss, and crushed leaves of Tetradenia riparia Hochst Codd, a traditional Rwandan medicine, were mixed with beans, Phaseolus vulgaris L., for storage protection. These plant-protected beans were compared with "off the shelf' beans that were being sold to consumers by the Rwandan National Agricultural Products Marketing Organization (OPROVIA). A trained sensory panel determined that beans treated with neem and C. cinerariaefolium were as acceptable after 8 months storage as those being sold throughout Rwanda by the marketing organization. Beans marketed by this organization were all treated with the standard insecticide application in Rwanda, 0.01% weight/weight pirimiphos methyl in a powder formulation. Instrumental hardness (% hard-to-cook/mean gram force) after 20 months of storage was acceptable for beans stored with neem or with C. cinerariaefolium or with the conventional government application of pirimiphos methyl. Use of either neem or C. cinerariaefolium for storage protection should not affect consumer acceptance of dry beans. PMID- 8719735 TI - Effects of various water or hydrothermal treatments on certain antinutritional compounds in the seeds of the tribal pulse, Dolichos lablab var. vulgaris L. AB - Effects of soaking, cooking and autoclaving on changes in polyphenols, phytohaemagglutinating activity, phytic acid, hydrogen cyanide (HCN), oligosaccharides and in vitro protein digestibility were investigated in seeds of Dolichos lablab var. vulgaris. Both distilled water and NaHCO3 solution soaking and autoclaving significantly reduced the contents of total free phenolics (85 88%) compared to raw seeds. Autoclaving (45 min) reduced the content of tannins by upto 72%. Soaking seemed to have limited effect in eliminating phytohaemagglutinating activity, whereas autoclaving (45 min) seemed to eliminate the haemagglutinating activity completely. The reduction in content of phytic acid was found to be some what greater in distilled water soaking (28%) compared to NaHCO3 solution soaking (22%). Only a limited loss in content of phytic acid was observed under cooking as well as autoclaving. Loss of HCN was greater under autoclaving (87%) compared to the other processes studied. Of the three sugars analysed, soaking reduced the level of verbascose more than that of stachyose and raffinose. Autoclaving reduced the content of oligosaccharides more efficiently (67-86%) than ordinary cooking (53-76%). Autoclaving improved the in vitro protein digestibility (IVPD) significantly (13%). Of all the different water and hydrothermal treatments studied autoclaving seemed to be the most efficient method in improving IVPD and eliminating the antinutrients investigated except phytic acid. PMID- 8719736 TI - The effect of solvent treatment on the chemical composition and organoleptic acceptability of traditional condiments from Nigeria. AB - Three Nigerian condiments from locust bean, melon, and soya bean were prepared by the traditional technique of uncontrolled fermentation and then partly defatted by hexane and di-ethyl ether extraction respectively. Proximate analysis and consumer preference tests were conducted. Results showed that while crude protein, ash and fibre contents remained virtually unchanged, the carbohydrate contents of the treated condiments (derived by difference) increased remarkably. The increase was associated with the significant reduction of about 50% (p = 0.05) in lipid contents of the three condiments. The observed solvent effect correlated positively with panelists' preference rating for the treated locust bean and melon condiments. Except with the soya bean condiment, higher mean scores were observed after the solvent treatment for the four sensory attributes assessed. However, condiment-type and treatment notwithstanding, colour and odour appear to critically determine the level of acceptability of condiments. PMID- 8719738 TI - Chemical and biological characteristics of a west African weaning food supplemented with compea (Vigna unguiculata). AB - The effects of cowpea and amino acid supplementation on the protein quality and chemical characteristics of a maize-based West African traditional weaning food were studied. Process optimization for improved nutritive value was also determined. Supplementation of the traditional weaning food with cowpea increased the lysine, tryptophan and threonine content while the sulphur-amino acids decreased with increasing levels of cowpea. Further supplementation of 70:30 maize/cowpea blends with lysine, threonine or methionine did not significantly improve (p > 0.05) protein quality in terms of the biological value (BV) and net protein utilization (NPU), although significant increases in the protein scores were noted. However, considerable improvements in the BV and NPU were recorded in blends fortified with either tryptophan alone or a combination of lysine, tryptophan, methionine and threonine. Cooking whole cowpea seeds for 45 min before incorporating in the blend formulation also significantly improved (p < 0.05) the protein quality of maize/cowpea blends. The BV and NPU of blends containing 30% pre-cooked cowpea increased from 52 to 76% and 50 to 71% respectively compared to pure maize porridge. The protein content increased from 10 to 14% and the utilizable proteins more than doubled. The weight increase of experimental rats fed with these blends was comparable to that of rats on casein diet. A 30% supplementation of the maize-based weaning food with cowpea therefore greatly enhances the nutritive value especially when the cowpea is pre-cooked for 45 min. Use of chemical scores alone for such blends cannot be a reliable index of blend quality. PMID- 8719737 TI - Digestibility of dietary fiber components in vegetarian men. AB - Digestibility of fiber components namely neutral detergent fiber (total content of cellwall) cellulose, hemicellulose and lignin are estimated in 14 healthy vegetarian men during adlibitum feeding and at 3 energy levels namely 2526, 2868 and 3290 kcals/day. Values of digestibility for adlibitum experiments were 34.17 +/- 2.3 for neutral detergent fiber (NDF), 30.1 +/- 3.9 for cellulose and 53.4 +/ 3.0 for hemicellulose and 8.1 +/- 2.6 for lignin. There was a considerable variability in digestibility of fiber components between individuals. PMID- 8719739 TI - Certain functional properties of gums derived from some lesser known tropical legumes (Afzelia africana, Detarium microcarpum and Mucuna flagellipes). AB - Gum was extracted from some lesser known tropical legumes--Afzelia africana, Detarium microcarpum and Mucuna flagellipes. The extracts were evaluated for some functional properties. At a constant shear rate the apparent viscosity of the gums was directly proportional to the gum concentration. At 2% concentration, the gum dispersions showed an apparent viscosity of 41.24, 115.05 and 145.14 cps for A. africana, D. microcarpum and M. flagellipes gums respectively, measured at 174/sec and 25 +/- 1 degree C with the NV sensor of Haake rotovisko viscometer. All the gums were found to contain D-galactose as a major monosaccharide. In addition, the presence of D-mannose, and D-glucose were indicated in both D. microcarpum and M. flagellipes while L-rhamnose, was indicated only in A. africana. A. africana showed significantly (p < 0.05) lower water absorption capacity and gelation properties than D. microcarpum and M. flagellipes. All the gums showed good emulsion properties. Emulsions of M. flagellipes gum showed little response to pH changes while A. africana and D. microcarpum gum showed better emulsion properties at acid pH than at alkaline pH. PMID- 8719740 TI - Effect of home processing on total and extractable calcium and zinc content of spinach (Spinach oleracia) and amaranth (Amaranthus tricolor) leaves. AB - Spinach (Spinacia oleracia) and amaranth (Amaranthus tricolor) leaves were stored in polyethylene bags and without packing for 24 and 48 hours in a refrigerator at 5 and 30 degrees C in polyethylene bags. The fresh leaves were also dried (oven and sun); blanched (5, 10 and 15 min) and cooked in an open pan and a pressure cooker. The processed leaves were analysed for total and extractable calcium and zinc content. The Ca and Zn content of these leaves varied from 1320 to 2120 and 11.70 to 12.60 mg/100 g DM and the percentage HCl-extractability was 77.82 to 81.92 and 85.16 to 86.15, respectively. No significant effects of drying and storage were observed on total Ca and Zn content and HCl-extractability while blanching and cooking resulted in significant improvement of HCl-extractability of these two minerals. Thus, cooking and blanching are good ways to improve the HCl-extractability of Ca and Zn. PMID- 8719741 TI - Physico-chemical properties and acceptability of yam flour substituted with soy flour. AB - Yam flour was substituted 10, 20 and 40% with defatted and full fat soy flour. The effect of the substitution on the proximate composition, swelling power, solubility, water binding capacity and Brabender visco amylograph cooking properties of the yam flour and acceptability of the cooked paste (amala), were evaluated. Protein contents of the mixtures were 23.0 and 25.5% on substituting 40% full-and defatted soy flours for yam flour, ash and crude fibre contents increased while carbohydrate content, swelling power, Brabender paste viscosities decreased with increase in soy flour substitution of yam flour. Colour, texture, taste and overall acceptability of pastes (amala) from the mixed flours were rated lower than that of yam flour. Up to 10% defatted and 20% full fat soy flour substitution for yam flour was acceptable for amala. PMID- 8719742 TI - Purine and pyridine nucleotide metabolism in the erythrocytes of patients with Rett syndrome. AB - The possible involvement of purine and pyridine metabolism in Rett syndrome, a neurodegenerative disorder of unknown aetiology affecting females, was investigated. The levels of purine and pyridine nucleotides and their metabolites were determined by HPLC in the erythrocytes and plasma of 31 Rett patients and of 17 age-matched controls. Nucleotide production rate from extracellular precursors was determined in intact cells and enzyme activities were assayed in crude lysates using the same HPLC method. Decreased plasma nicotinamide concentrations and lower erythrocyte activities of hypoxanthine phosphoribosyl transferase, adenine phosphoribosyl transferase and phosphoribosylpyrophosphate synthetase were observed in Rett children compared with age-matched controls, while the production rate of IMP from hypoxanthine and of total pyridine nucleotides from nicotinic acid by intact erythrocytes was significantly increased. No significant difference was found in any of the other parameters examined. These findings give a new contribution to the knowledge of the biochemical alterations in Rett syndrome and encourage further investigations in the nucleotide field. PMID- 8719743 TI - Deficiency of merosin (laminin M or alpha 2) in congenital muscular dystrophy associated with cerebral white matter alterations. AB - Recently, a deficiency of merosin has been reported in patients with classical congenital muscular dystrophy (CMD), while other patients, with indistinguishable clinico-pathological features, do not present this deficiency, suggesting genetic heterogeneity. The purpose of the present investigation was to assess merosin distribution and quantity in 21 clinically well characterized Brazilian CMD patients, in order to: a) estimate the proportion of merosin-deficient cases in this group of patients; b) characterize phenotypically merosin-negative, as compared to merosin-positive patients. Merosin deficiency was found in 11 patients and all the seven who had been submitted to neuroimaging studies showed evidence of periventricular dysmyelination. A normal pattern of 43 DAG was found in all patients, which suggest that this protein is not preferentially involved in a third form of merosin-positive CMD. Results from the present study are further suggestive, but do not prove, that the association of merosin deficiency with white matter alterations represents a genetic entity with common clinical, laboratory and neuroimaging findings. PMID- 8719744 TI - Neuroradiological and eye movement correlates in children with intermittent saccade failure: "ocular motor apraxia". AB - Ocular motor apraxia (OMA) is a clinical sign involving the intermittent inability to initiate saccades, and a failure of quick phases during optokinetic nystagmus (OKN) and vestibular nystagmus (VN). Some patients have no other associated abnormalities (idiopathic), whereas others have a variety of neurological conditions. We quantified the severity of the saccade failure and correlated it with neuro-radiological and other oculomotor findings in 62 children (aged 17 days - 14 years). Saccades, smooth pursuit, OKN and VN were recorded using electrooculography and the extent of "locking up" (absent quick phases during OKN and VN) was measured. Saccades were usually hypometric. Pursuit and OKN gains were normal in the majority of the idiopathic cases but were low in those with other neurological conditions. Twenty-four patients had essentially normal scans, whereas 38 had abnormal scans: Delayed myelination, cerebellar abnormalities (particularly involving the vermis), and agenesis of the corpus callosum were the most common findings. A significant positive correlation was present between increasing neuro-radiological deficits and severity of "locking up" during OKN. Principal component analysis showed that brainstem and cerebellar vermis abnormalities were the main factors involved. A pathophysiological basis of OMA is discussed in the light of animal and clinical studies. PMID- 8719745 TI - Progressive facial hemiatrophy with multiple benign tumors and hamartomas. AB - We report the unusual association of Progressive Facial Hemiatrophy (Parry Romberg syndrome) with multiple benign tumors (orbital neurinoma, mandibular odontogenous fibroma) and hamartomas. The neurological clinical features were infantile hemiplegia, mild mental retardation and focal seizures. Brain CT-scan and MRI showed porencephaly and cerebral calcifications ipsilateral to hemifacial atrophy. Immunological investigations proved negative. The etiology of the disease and the bridging of this case to phakomatoses are discussed. PMID- 8719746 TI - Impaired peripheral somatosensory function in children with Down syndrome. AB - Sensory neurography of the median nerve was performed bilaterally in 6 children with Down syndrome (DS) aged 11-16 years and in 10 healthy controls of similar age. The sensory nerve conduction velocities from the thumbs and 3rd fingers were recorded and were significantly subnormal in the DS group. Sensory nerve action potentials were also lower in the DS children and the difference was particularly evident following stimuli of the thumbs. Using scalp electrodes sensory evoked potential latencies were recorded in five of the DS children and all controls. There was no obvious difference between the subjects with DS and the controls. It is suggested that an impaired peripheral somatosensory function should be added to previously known symptoms constituting DS. The results may contribute to previous findings of a poor performance of DS children in tests of tactual perception. PMID- 8719747 TI - Familial childhood primary lateral sclerosis with associated gaze paresis. AB - Three children from consanguineous parents began losing the ability to walk in late infancy. Despite chronically progressive weakness leading to wheelchair dependence by adolescence and later loss of motor speech production, intellect remained preserved. Examination revealed upper motor neuron findings of pseudobulbar palsy and spastic quadriplegia, without dementia, cerebellar, extrapyramidal or sensory signs. In addition they exhibited a diffuse conjugate saccadic gaze paresis, especially severe on down-gaze. CT and MRI scans of brain and spinal cord, EEGs, visual and brainstem auditory evoked potentials, CSF examinations, enzyme assays for lysosomal storage diseases, blood amino acids and urine organic acids were all normal. Cortical somatosensory evoked potentials were poorly configured in two of the patients, though they had normal central conduction. EMG showed no signs of denervation. Nerve conduction studies showed normal peripheral motor and sensory conduction velocities. Transcranial magnetic stimulation of the brain elicited no motor-evoked potentials. Despite the lack of neuropathological confirmation, the clinical course and neurophysiologic data strongly support the diagnosis of a familial (autosomal recessive) primary lateral sclerosis (PLS). PMID- 8719748 TI - Joubert syndrome: are kidneys involved? AB - Renal involvement was checked in our series of 12 children with Joubert syndrome (JS) who fulfil the cardinal diagnostic features. No patient had clinical evidence of congenital retinal dystrophy. One child had normal kidneys at autopsy and 11 children (aged 1 month to 15 years) had no evidence of cystic kidney changes on ultrasonography. We conclude that kidneys are not involved in JS without retinal dystrophy. There may be a subgroup (or different syndrome) of patients with both renal cysts plus retinal dystrophy, as suggested by Saraiva and Baraitser (11). PMID- 8719749 TI - A mild form of infantile isolated sulphite oxidase deficiency. AB - Neonatal sulphite oxidase deficiency is characterised by severe neurologic dysfunction, brain atrophy, dislocation of the lens and increased urinary excretion of sulphite, thiosulphate, taurine and S-sulphocysteine, and by a low plasma cystine. We present clinical, neuroradiological and biochemical data of a patient with late onset symptoms comparing this presentation with the neonatal form and stressing the difficulties of the diagnosis of this disorder. PMID- 8719750 TI - Fucosidosis with dystonia. AB - Fucosidosis, a progressive neurodegenerative disease, evident in early childhood, is associated with progressive loss of mental and motor function and increasing spasticity and hyperreflexia. We report a Canadian male, with clinical features similar to previously reported fucosidosis patients, however, since age 5 he has exhibited progressive dystonic posturing, initially unilateral, but recently involving both lower limbs. Extensive study of his cultured lymphoblasts demonstrated that alpha-fucosidase activity and immunoreactive alpha-fucosidase protein were absent. He is homozygous for the Q422X mutation, a C to T transition within exon 8 of the alpha-fucosidase gene which results in loss of an EcoR1 restriction enzyme cut site. Even among the 4 other reported fucosidosis families having one or more individuals homozygous for this same (Q422X) mutation there was no previous report of dystonia. PMID- 8719751 TI - Transient neonatal nonketotic hyperglycinemia: a 13-year follow-up. AB - We report on a patient with transient nonketotic hyperglycinemia (NKH). Onset, clinical features, biochemical and electroencephalographic findings were consistent with neonatal nonketotic hyperglycinemia. The normalization of clinical, biochemical and electroencephalographic features on day 9 without specific therapy suggested a transient form. At the last follow-up the girl was 13 years old, had a normal mental development, no EEG abnormalities or neurological deficits. PMID- 8719752 TI - Progressive subcutaneous and periarticular calcifications in a patient with spinal muscular atrophy type III. PMID- 8719753 TI - Analysis of a human brain voltage-gated potassium channel gene, KCNA6 (HBK2), in patients with juvenile myoclonic epilepsy. PMID- 8719754 TI - Acetylcholine receptor antibodies and single-fiber EMG in first-degree relatives of children with myasthenia gravis. PMID- 8719755 TI - Non-reciprocal connections between the perihypoglossal nuclei and the cerebellar paramedian lobule in rabbits and their relationship to the climbing fiber zones. AB - Retrogradely transported horseradish peroxidase (HRP) was used to investigate the organization of projections from the perihypoglossal nuclei to the cerebellar paramedian lobule, and possible reciprocal corticoperihypoglossal connections in rabbits. The absence of labeled Purkinje cells in the paramedian lobule after HRP injections of the perihypoglossal nuclei, compared with labeling in the nodulus, ipsilateral flocculus and adjacent ventral paraflocculus, strongly suggests that corticoperihypoglossal projections do not originate in the paramedian lobule. Comparison of labeling patterns in the perihypoglossal nuclei following HRP injections into various sublobules of the paramedian lobule suggests that projections from the perihypoglossal nuclei to the paramedian lobule exist and that some degree of topographic organization is present exclusively in bilateral projections from the caudal part of nucleus prepositus hypoglossi. All perihypoglossal projections end in deep parts of sublobules e-b of zones C2 and D1 (as defined by inferior olivary labeling). It is significant that in the ipsilateral nucleus prepositus hypoglossi, two groups of cells projecting to zone C2 are arranged rostrocaudally between three other groups projecting to zone D1. PMID- 8719756 TI - The dermatomes of the pectoral and pelvic fin of the long-tail stingray Himantura fai. AB - Recorded action potentials in whole spinal nerves during mechanical stimulation of the skin of Himantura fai revealed that sequential nerves innervated sequential overlapping strips of the pectoral and pelvic fin skin. As found in previous studies, in which the dermatomes of the skate Raja clavata were measured behaviourally, approximately one-third of the rostral and caudal regions of each dermatome overlapped with the adjacent dermatomes. Consistent with dermatomal maps from non-mammalian vertebrates, but unlike the dermatomal maps obtained in mammals, there appears to be little difference in dermatome size when measured behaviourally or electrophysiologically. We suggest that neural mechanisms of the spinal cord, which appear to contribute to the discrepancy between behaviourally and electrophysiologically mapped dermatomes in mammals, are of negligible influence in stingrays. PMID- 8719757 TI - Hair cell heterogeneity in the goldfish saccule. AB - A set of cytological studies performed in the utricle and saccule of Astronotus ocellatus (Teleostei, Percomorphi, Cichlidae) identified two basic types of hair cells and others with some intermediate characteristics. This paper reports on applying the same techniques to the saccule of Carassius auratus (Teleostei, Otophysi, Cyprinidae) and demonstrates similar types of hair cells to those found in Astronotus. Since Carassius and Astronotus are species of extreme taxonomic distance within the Euteleostei, two classes of mechanoreceptive hair cells are likely to represent the primitive condition for sensory receptors in the euteleost inner ear and perhaps in all bony fish ears. PMID- 8719758 TI - The population of GnRH-containing neurons showing socially mediated size changes project to the pituitary in a teleost, Haplochromis burtoni. AB - Reproductive function in all vertebrates is controlled by the brain-pituitary gonadal axis. In teleost fish, endocrine cells within the adenohypophysis are grouped together and each collection of cells is innervated by specific neuropeptide fibers. An important regulatory step in reproductive control is gonadotropin-releasing hormone (GnRH), whose delivery to the pituitary is responsible for its release of gonadotropins. The hormone GnRH has been shown to play a critical role in the social control of reproduction in a teleost fish, Haplochromis burtoni. However, there has been no direct evidence that the preoptic area GnRH neurons project to the pituitary. In this study, we used a retrograde tracer and immunohistochemistry to identify those GnRH containing neurons that project to the adenohypophysis. We compared reproductively active territorial males with quiescent non-territorial males to discover whether the connectivity of the preoptic area GnRH neurons depends on the reproductive status of the male. We found that, irrespective of reproductive status, most GnRH neurons in the preoptic area project to the pituitary and that all of these GnRH neurons show the soma size change that has been associated with reproductive status in Haplochromis burtoni. Based on these data, we propose that there is a single population of GnRH containing cells in the preoptic area that change size as a function of reproductive state and that this entire population projects to the pituitary. This is the first direct demonstration that this essential circuit, linking GnRH neurons in the preoptic area to the pituitary, exists. PMID- 8719759 TI - The extrabulbar olfactory pathway: primary olfactory fibers bypassing the olfactory bulb in bony fishes? AB - Recent evidence has revealed that some primary olfactory fibers bypass the olfactory bulb and terminate in tel- and/or diencephalic areas (extrabulbar olfactory pathway, EBOP). We investigated the projections of this system in different fishes by means of soybean agglutinin binding studies. In all species in which primary olfactory fibers were labelled, fiber bundles can be traced beyond the olfactory bulb. These run with the medial forebrain bundle and terminate at different targets, depending on the species. In the teleosts Macrognathus, Mogurnda, and Hemichromis, EBOP fibers can be traced into the ventral telencephalon, pars ventralis, pars supracommissuralis and/or into the preoptic nucleus. In most nonteleosts studied (Polypterus, Chalamoichthys, Amia), the EBOP also innervates diencephalic targets. An exception is Acipenser, which displays an innervation pattern similar to that in teleosts. Comparison with results obtained by other techniques suggests that the EBOP consists of primary olfactory fibers, which project not only to the olfactory bulb but also to various other targets in the prosencephalon of anamniotic vertebrates. PMID- 8719760 TI - Dialysis prescription. PMID- 8719761 TI - End-stage renal disease in the USA: data from the United States Renal Data System. AB - Treated end-stage renal disease continues to increase at an alarming rate in the US. There has been an exponential growth in the incidence rate between 1982 and 1991 at the rate of 8.76% per year. Approximately 218,042 patients received treatment for ESRD in 1991, of which 49,909 were new patients. Although the increase in the incidence rate is seen for all the major disease categories responsible for ESRD, diabetes mellitus, probably type 2, and hypertension are responsible for the bulk of the increase. African Americans and Native Americans have shown the most dramatic increase; diabetes being the major reason for both races, but for African Americans, hypertension is the leading cause of ESRD. A bulk of the increase in the ESRD patient population has been in the older age (greater than 65 years of age) group. The mortality rate for the ESRD patient population, and, specifically, for the dialysis population remains relatively high, with 1-year survival probabilities of approximately 78%. Some of the contributing factors cited for the high death rate, especially in the dialysis patient population include inadequate dialysis dose, low flux of the dialysis membranes, shortened dialysis times, an increase in the age of the ESRD population, and bioincompatible dialysis membranes. The effect of the widely practiced dialyzer reuse on dialysis patient morbidity and mortality remains unclear. PMID- 8719762 TI - Quantifying hemodialysis. AB - The interpretation of traditional serum urea and creatinine concentrations as indices of the severity of uremia requires major modifications in hemodialyzed patients. Although high urea concentrations usually signify worsening uremia and inadequate dialysis, low concentrations do not guarantee a good outcome. Urea production as modified by diet and other factors must also be included in a complete description of dialysis quantity and adequacy. The expression 'Kt/V' is a measure of hemodialysis that includes both urea removal and urea generation and is easy to measure from predialysis and postdialysis serum urea concentrations. Kt/V can be most precisely measured with the aid of mathematical models of urea kinetics during and between hemodialyses. Although a reliable measure of the dialysis dose received by most patients, the single-compartment model overestimates serum urea concentrations during hemodialysis and fails to predict the rebound immediately following dialysis. The classic two-compartment model that includes a factor for resistance to diffusion between the compartments, more accurately predicts the BUN profile but fails to account for blood flow-related disequilibrium including cardiopulmonary recirculation. Since solute disequilibrium reduces the effectiveness of hemodialysis, models that incorporate equilibrated urea concentrations both before and after hemodialysis are potentially more accurate tools for quantifying dialysis. Dialysate methods have the potential to accurately measure both solute removal which is the ultimate goal of dialysis, and patient clearance which is considered a better measure of the dialysis effect than dialyzer clearance. Application of these newer techniques requires major changes in sampling methods and changes in analytical equipment that will delay implementation. Meanwhile, analysis of blood-side urea concentrations using the single-compartment, variable volume model provides a reasonable estimate of Kt/V but must be interpreted with due consideration of its well-recognized pitfalls. PMID- 8719763 TI - Impaired delivery of dialysis: diagnosis and correction. AB - Hemodialysis treatments yielding inadequate amounts of dialysis, as defined by urea kinetic modeling, are partially responsible for considerable mortality and morbidity in the United States. In almost 50% of dialysis treatments resulting in a Kt/V of < 1.0, the culprit is impaired delivery of the prescribed amount of dialysis. The factors involved in impaired delivery of dialysis are many and often elusive. If present and widespread, a search for the cause of the problem entails careful examination of the equipment and nursing procedures. If impaired delivery is a sporadic and infrequent event, a patient-specific investigation should be undertaken. In either circumstance, a clear understanding of the principles and practical aspects of hemodialysis greatly assists the nephrologist as a sleuth. PMID- 8719764 TI - Importance of treatment time and blood pressure control in achieving long-term survival on dialysis. AB - A drastic reduction in hemodialysis (HD) time has been based on the dialysis dose measurement in terms of urea Kt/V exclusively and on the use of high efficiency dialysers. It was subtly coupled to a de-emphasis of the use of HD to normalize blood pressure. In Tassin we have maintained long slow HD with an overall excellent patient survival. We analyze the influence of the different factors of this survival: the place of dry weight and blood pressure control without use of antihypertensive medication is emphasized, and the role of dialysis dose and nutrition is discussed. Adequacy should be defined in terms of these additive conditions. Long slow HD allows one to fulfill these conditions easily. Shortening of dialysis time should not interfere with the control of blood pressure. PMID- 8719765 TI - Hemodialysis vascular access: effect on urea kinetics and the dialysis prescription. AB - The effective delivery of dialysis requires repeated reliable access to the central circulation capable of providing rapid blood flow. This access to the circulation continues to be the 'weak link' in the provision of long-term renal replacement therapy. Dialysis access malfunction is a major cause of inadequate dialysis delivery and venous stenosis is the leading cause of access malfunction and thrombosis. Careful monitoring of venous dialysis pressures and recirculation along with urea kinetic modeling and physical examination of the graft can prospectively identify the malfunctioning vascular access. When these indicators are used for referral for fistulogram, venous stenosis can be identified and corrected before graft thrombosis. Not only can preemptive repair of the vascular access prevent thrombosis, it also allows for more efficient dialysis delivery to the patient. PMID- 8719766 TI - The dialysis prescription: reuse. AB - Despite the widespread long practice of reuse there is continued criticism of the technique both in the media, journals and more recently at specially convened meetings. In the United States there are three principal sterilant/germicides used in reprocessing hemodialyzers: peracetic acid, formaldehyde and glutaraldehyde. The use of peracetic acid now exceeds that of formaldehyde while the more recent development of heat sterilization is only practiced in a few units. The major advantages of reprocessing include reduced cost, reduced first use syndrome and intradialytic symptoms which may be a result of increased biocompatibility with reuse. There is little data with regard to hospitalization and the recent reports of mortality rates are discussed. The major disadvantages associated with reprocessing include risks of infection, chronic exposure and accidental exposure to patients and staff to the sterilant germicide. We conclude that the reuse of dialyzers continues to be appropriate provided units follow the manufacturer's guidelines, the AAMI Recommended Practices and have a method in place to ensure the prescribed dialysis prescription is delivered. PMID- 8719767 TI - Water treatment for hemodialysis. AB - Water treatment is a vital aspect of hemodialysis in which knowledge and technical skills are of utmost importance. The recognition that nontuberculous mycobacteria can be resistant to certain germicides spurred the establishment of the current safety microbiologic standards for dialyzer reprocessing. Monitoring the dialyzer membrane integrity is as important as meeting the standards for bacterial and endotoxin levels for dialyzer reprocessing. Ensuring the use of product water that meets the chemical and microbiologic standards of the Association for the Advancement of Medical Instrumentation is necessary to reduce the incidence of endotoxemia and chemical hazards associated with the use of water for hemodialysis. The pathogenesis of febrile reactions during hemodialysis remains controversial. The weight of evidence, however, favors transmission of endotoxin fragments across dialysis membranes to induce mononuclear cell cytokine production. PMID- 8719768 TI - Injectable medications for hemodialysis patients. AB - Increasing use of medications designed specifically for intravenous administration during hemodialysis has dramatically improved the treatment of many comorbid conditions accompanying end-stage renal disease. Recombinant human erythropoietin has resulted in definitive treatment of the hypoproliferative anemia of uremia, while sustained response to this hormone has been facilitated by treatment with intravenous iron dextran. Simple and effective therapies for many gram-positive infections in end-stage renal disease patients have been realized through the favorable pharmacokinetics and low toxicity of vancomycin when administered during hemodialysis. Major improvements in the therapy of high turnover bone disease have resulted from the increasing use of intravenous calcitriol during dialysis as well. The contributions of these four medications in improving patient care and the benefits of their administration during the dialysis procedure are reviewed. PMID- 8719769 TI - Nutrition and the dialysis prescription. AB - Malnutrition is common among patients with acute and chronic renal failure. The efficiency of modern dialytic techniques has allowed for more liberal administration of nutrients to patients with renal failure, particularly with regard to protein and amino acids. Protein restriction is not indicated for patients on dialysis, and should be employed cautiously, if at all, in patients with renal insufficiency. The 'nutrition prescription' should be considered a vital part of the comprehensive medical, surgical, and dialytic care provided to patients with renal disease. PMID- 8719771 TI - A congenital heart defect in Drosophila caused by an action-potential mutation. AB - The mutation no action potential (nap) induces arrhythmia in the heartbeat of Drosophila melanogaster larvae at temperatures above 20 degrees C; heartbeat becomes normally rhythmic again after a shift back to 20 degrees C. For this phenotype, napa is almost completely recessive to the wild type, napa also reduces the temperature-sensitivity of heart rate over a wide range of temperature, for this phenotype, napa is dominant over the wild type, napa causes reversible paralysis in adults by epistatic effects on the expression of paralyrica, a gene encoding a voltage-dependent sodium channel. However, the paramutation, which induces paralysis in adults at 29 degrees C, has no effect on larval heartbeat at temperatures between 20 degrees and 37.5 degrees C. The period gene, contra earlier reports, has no effect on heartbeat. PMID- 8719770 TI - Distinct mechanisms of action of the Lozenge locus in Drosophila eye and antennal development are suggested by the analysis of dominant enhancers. AB - The development of the olfactory sense organs on the antenna of the fruit fly Drosophila utilises mechanisms distinct from those used in the rest of the adult peripheral nervous system. Lozenge (lz) is the only locus hither-to identified as required for the development of antennal sense organs. In addition to effects on the antenna, mutations in lz also affect the development of the eye and maxillary palp. We have used the readily-scored eye-phenotype in a temperature sensitive lz allele to screen for dominant modifiers of phenotypes at this locus. We analyse the phenotypes of both intragenic and extragenic modifiers. Our results reinforce the view from developmental studies that lz functions in eye and antennal development in distinct ways. PMID- 8719772 TI - Two isoforms of Drosophila dynamin in wild-type and shibire(ts) neural tissue: different subcellular localization and association mechanisms. AB - The temperature-sensitive mutations of the shibire (shi) gene in Drosophila cause endocytic arrest, resulting in neurotransmission block and paralysis at high temperatures. However, underlying mechanism for the defects is not yet known. We examined the subcellular distribution of dynamin, a product of the shi gene, by immunoblotting and immunocytochemical assays. Two isoforms of dynamin with apparent M(r) of 92 kD and 94 kD have been detected in wild-type and shi(n) adult neural tissue. The two isoforms were reproducibly associated with different subcellular fractions of head homogenates. The 94kD isoform is fractionated in the low speed (2.000 x g) pellet containing plasma membrane fragments, and the 92kD isoform in the high speed (130,000 x g) pellet. In this procedure, very little dynamin remained in the high speed supernatant fraction. The 94 kD isoform represents the majority (65-75%) of total dynamin and appears to be a peripheral membrane protein. It can be extracted from the low speed membrane pellet by high salt, Na2CO3 (pH 11) or Triton X-100 treatments. Extracted 94kD dynamin from both wild-type and mutant homogenates is able to reassociate with artificial phospholipid vesicles at both permissive and restrictive temperatures. Binding of the 94 kD dynamin to liposomes appears to be pH-dependent, varying most significantly within the physiological pH range, which may be functionally important. The 92 kD isoform cannot be released by high salt or Na2CO3 treatments and only a small fraction is released by Triton X-100, suggesting a different mechanism of association with cell structures. The distribution of the two isoforms is not altered by the presence of stabilized microtubules in homogenates. No apparent degradation or subcellular redistribution of mutant dynamin was detected in two shi(n) alleles after heat shock or block of the dynamin GTPase activity, suggesting that intracellular redistribution or degradation of mutant dynamin are not involved in the endocytosis arrest in these mutants. These observations resemble the effect of endocytosis arrest by GTP gamma-S in rat brain synaptosomes (Takei et al., 1995), in which dynamin is trapped at the neck of invaginated pits but is absent in the clathrin-coated distal end undergoing internalization. Our finding that endocytosis arrest by shi(n) mutations and GTP-gamma-S do not lead to cumulation of dynamin in the low speed pellet fraction further suggests that the 94 kD isoform remains associated with the plasma membrane during coated vesicle pinch-off and that the two isoforms do not appear to correspond to different functional states of dynamin but are likely to be involved in separate cellular compartments within the membrane cycling pathway (e.g., the plasma membrane, endosomes, and endoplasmic reticulum). PMID- 8719773 TI - Renin gene expression in the aging kidney: effect of sodium restriction. AB - The activity of the renin-angiotensin system as well as the ability of the kidney to retain sodium following salt restriction are reduced with age. The relationship between these age-related changes in renal function and the renin gene expression was presently investigated. The concentrations of renin and its mRNA were measured in kidney of 10- and 30-month-old control female WAG/Rij rats and of animals which were salt restricted for 4 days. In the senescent rats, the kidney renin concentration, like the plasma concentration of angiotensin II, was half that in adult rats. The intrarenal content of renin mRNA did not differ between 10- and 30-month-old animals, suggesting that the transcriptional rate of the renin gene is unchanged with age. During the early phase of adaptation to sodium depletion, the systemic angiotensin II concentration was not modified in either age groups. Four-days salt restriction did not significantly change the renal storage of renin. In contrast, this short term salt restriction induced a 2.3-fold increase in the renin mRNA in adult kidney, and a 1.9-fold increase in the senescent kidney. These data suggest that the age-related decrease in renal concentration of renin is linked to a modification in the rate of translation of renin mRNA, or to an alteration in the protein maturation. The difference in adaptation to the early phase of salt restriction with age should not be linked to changes in renin gene transcription, but more likely to a change in the tissue response to the local renin-angiotensin system. PMID- 8719774 TI - Altered regulation of pituitary luteinizing hormone secretion by GnRH and inhibin in the aged persistent-estrous female rat. AB - The physiologic regulation of pituitary LH secretion is under the direct control of hypothalamic GnRH as well as feedback control via ovarian steroid and peptide hormones. Changes within the hypothalamic-pituitary-ovarian axis during reproductive aging lead to the gradual attenuation and the eventual loss of cyclic LH release that is seen in the old persistent-estrous rat. In the following studies, we determined whether changes in the pituitary response to regulation by hypothalamic GnRH and/or ovarian inhibin could be involved in age related deficits in LH release by examining the direct effect(s) of GnRH and inhibin on LH release in old persistent-estrous (PE) and young female rats using an in vitro pituitary cell culture system. Both pituitary LH content and LH release were significantly lower in PE pituitary cells compared to young pituitary cells. GnRH (10 ng/ml) significantly increased LH release above basal levels in young and PE pituitaries. However, the magnitude of LH release was significantly lower in PE pituitaries compared to young pituitaries. Treatment with recombinant inhibin (0.1, 1, or 5 ng/ml) did not significantly affect basal or GnRH-stimulated LH release from young rat pituitaries under our culture conditions. In contrast, there were significant decreases in the levels of both basal LH and GnRH-stimulated LH release from PE pituitaries after treatment with 1 or 5 ng/ml human recombinant inhibin, as well as with inhibin plus 10 ng/ml GnRH, respectively. These results suggest that there are significant changes in the pituitary response to GnRH and ovarian inhibin regulation that may lead to significant alterations in LH secretion during aging. PMID- 8719775 TI - Vitamin A incorporation into lipofuscin-like inclusions in the retinal pigment epithelium. AB - Intravitreal injection of the protease inhibitor leupeptin causes a rapid accumulation of lipofuscin-like autofluorescent inclusions in the retinal pigment epithelium (RPE) of the eye. In vitamin A-deprived animals, similar inclusions form in response to leupeptin treatment, but they do not become autofluorescent. Because vitamin A is necessary to the development of fluorescence, it appears likely that retinoids are directly incorporated into the inclusions. Experiments were conducted to determine whether this is the case. Rats were reared on a diet containing retinoic acid as the only retinoid. Retinoic acid cannot be utilized in visual transduction by the retina. When the eyes had been over 90% depleted of visual cycle retinoids, the animals were given a single intramuscular injection of 3H-all-trans retinol. After 7 days, when visual cycle retinoids had returned to an average of almost 70% of normal, the animals were given an intravitreal injection of leupeptin in each eye. At either 1 day or 7 days after the leupeptin treatment, some of the animals were dark-adapted for at least 12 h. The eyes were enucleated and fixed under dim red light. A region of each retina just superior to the optic nerve head was examined with electron microscopic autoradiography. At both one day and 7 days after the leupeptin treatment, the radiolabel in the RPE was primarily associated with the leupeptin induced inclusion bodies. Label was also present in the photoreceptor outer segments. The localization of vitamin A to the leupeptin-induced inclusions in the RPE strongly suggests that vitamin A is covalently bound to outer segment proteins that have been phagocytosed by the RPE but remain undegraded due to protease inhibition. This bound vitamin A is probably responsible for the autofluorescence of the leupeptin-induced inclusions. Vitamin A is not likely to be bound through a Schiff base linkage, since retinal-Schiff base compounds do not exhibit lipofuscin-like fluorescence. PMID- 8719776 TI - Age-related enhancement of tumor necrosis factor (TNF) production in mice. AB - We investigated age-related changes in the production of TNF at the cellular level using immunocompetent peritoneal and spleen cells from C3H/He mice of various ages. The density of cultured peritoneal macrophages and spleen cells required for TNF production was at least 5 x 10(5) cells/dish. The optimal concentration of OK-432 for 24-h culture of peritoneal macrophages (1 x 10(6) cells) and spleen cells (1 x 10(7) cells) was 0.5 and 0.1 KE/ml, respectively. Among peritoneal cells, adherent macrophages were the major TNF-producing cells, whilst nonadherent T or B cells alone did not produce TNF after stimulation with OK-432. In the case of spleen cells, T or B cells were involved in the production of TNF when cultured with a few adherent cells in the presence of OK-432. However, T or B cells alone failed to produce TNF. Production of TNF by peritoneal macrophages from both male and female mice increased significantly with aging. In contrast, although TNF production by spleen cells tended to increase with aging, no significant change was noted. The total number of peritoneal and spleen cells, respectively increased up to about 18 months after birth with B cells being principally responsible for this age-related increase. We previously reported that systemic production of TNF increases with aging. The present study of TNF production at the cellular level in mice indicated (1) that TNF production per macrophage increased with aging, and (2) that the number of T and B cells involved in the production of TNF in the presence of macrophages also increased at least up to middle age. PMID- 8719777 TI - Contents of D-lactate and its related metabolites as well as enzyme activities in the liver, muscle and blood plasma of aging rats. AB - As it is generally known. L-lactate is formed via the Embden-Meyerhof glycolytic pathway from triosephosphates, whereas D-lactate is formed via methylglyoxal in rat. In this paper, age-related changes in the levels of D-lactate and its related compounds in rat tissues are reported. Rats from 5 weeks to 30 months old were used in these experiments. (1) We observed that rats above 27 months old were decrepit as judged by external appearance movement and other physiological data of them. (2) The hepatic levels of D-lactate, methylglyoxal and pyruvate became markedly lower in aging rats, especially the D-lactate content in 30 month old rats was lower by 90% than that of the 5 week-old rats. (3) As for plasma, D lactate and phosphate levels became lower with aging, whereas levels of L-lactate and pyruvate were not altered. (4) In skeletal muscle, aging caused a lower methylglyoxal concentration. The D-lactate level was markedly decreased at the age of 30 months in muscle. (5) As for enzyme, activities of glyoxalase I and II became markedly decreased with age in livers, whereas the activity of glyoxalase I in muscle was maintained at control level and glyoxalase II increased with age. PMID- 8719778 TI - Alterations of superoxide dismutase iso-enzyme activity, content, and mRNA expression with aging in rat skeletal muscle. AB - The alterations of superoxide dismutase iso-enzyme (Cu,Zn-SOD and Mn-SOD) activities, contents, and mRNA expressions with aging were studied in rat soleus muscle (SO) and extensor digitorum longus muscle (EDL). The activity and content of Cu,Zn-SOD in both muscles were significantly higher in old rats (24 months old) than in young rats (4 months old), whereas those of Mn-SOD showed no difference between young and old rats. After normalization to citrate synthase (CS) activity, however Mn-SOD/CS ratio in SO also showed the age-related increase. Moreover, the activities of other major antioxidant enzymes, glutathione peroxidase (GPX) and catalase (CAT), indicated age-related increases only in SO. As for the expressions of mRNAs for SOD iso-enzymes, that of Cu,Zn SOD in either muscle showed no significant change with aging, unlike its activity and content, although that of Mn-SOD was decreased with aging only in EDL. Thus, aging appeared to raise the level of antioxidant enzyme system in rat skeletal muscle. However, the resistance of Cu,Zn-SOD and Mn-SOD to oxidative stress accompanied by aging was different, the former being obviously greater than the latter. Such changes also differed in muscle fiber type suggesting that fast twitch fibers are more susceptible to age-related oxidative stress than slow twitch fibers. PMID- 8719779 TI - Depletion of cytosolic GSH decreases the ATP levels and viability of synaptosomes from aged mice but not from young mice. AB - The effect of glutathione depletion on the viability of freshly isolated synaptosomes from whole brain was investigated in young and aged mice. Aging did not influence the GSH and ATP levels and the viability of these synaptosomes. However depletion of glutathione caused by the cytosolic glutathione inhibitor diethyl maleate (1 mM) resulted in a significant decline, after 60 min of incubation, in ATP levels and viability in the synaptosomes from aged mice but not in those from young mice. When synaptosomes were incubated in the presence of the mitochondrial glutathione inhibitor ethacrynic acid (0.2 mM) there was a similar decline in glutathione, ATP levels and synaptosomal viability, both in young and aged mice. These results emphasize the relative importance of the cytosolic glutathione pool for the maintenance of the plasma membrane integrity in synaptosomes from aged mice. PMID- 8719780 TI - Endothelium-dependent relaxation to the B1 kinin receptor agonist des-Arg9 bradykinin in human coronary arteries. AB - Des-Arg9-bradykinin (des-Arg9-BK) caused endothelium-dependent relaxations in human, isolated coronary arteries which upregulated with in vitro incubation time. Relaxations to des-Arg9-BK were inhibited by the B1 receptor antagonist, des-Arg9-[Leu3]-BK (pK(B), 6.14 +/- 0.11) but were unaffected by the B2 receptor antagonist, Hoe-140. Therefore, this is the first demonstration that human coronary arteries possess endothelial B1 receptors which mediate endothelium dependent relaxation and appear to be synthesized de novo during the incubation period. PMID- 8719781 TI - The effect of the desglycinyl metabolite of remacemide hydrochloride (FPL 12495AA) and dizocilpine (MK-801) on endogenous amino acid release from mouse cortex. AB - 1. In this study the effect of FPL 12495AA, the desglycinyl metabolite of remacemide hydrochloride and dizocilpine (MK-801), on potassium- and veratridine stimulated release of neurotransmitter amino acids from mouse cortical slices was investigated. 2. Veratridine (20 microM) and potassium (60 mM) produced a preferential release of glutamate and aspartate. Potassium-stimulated release was calcium-dependent, while veratridine-stimulated release was only partially affected by removal of calcium from the medium. 3. FPL 12495AA significantly inhibited veratridine- and potassium-stimulated release of glutamate and aspartate. Lower concentrations of FPL 12495AA were needed to inhibit veratridine stimulated release of glutamate (12.5 microM) than potassium-stimulated release (100 microM). 4. Dizocilpine significantly inhibited veratridine- and potassium stimulated release of glutamate and aspartate at concentrations of 100 microM and above. 5. FPL 12495AA and dizocilpine both have an affinity for the ion channel subsite of the N-methyl-D-aspartate (NMDA) receptor. The reduction of potassium stimulated release of glutamate and aspartate by FPL 12495AA and dizocilpine is probably due to NMDA receptor blockade. 6. FPL 12495AA inhibited veratridine stimulated release at a concentration of 12.5 microM while dizocilpine was effective only at a concentration of 100 microM. This difference in efficacy is probably due to the higher affinity of FPL 12495AA compared to dizocilpine at the veratridine-binding site on the sodium channel. PMID- 8719782 TI - Neuroprotective efficacy of lifarizine (RS-87476) in a simplified rat survival model of 2 vessel occlusion. AB - 1. A new, modified rat two vessel occlusion model (with hypotension) was established and the neuroprotective efficacy of the novel agent lifarizine (RS 87476) was examined. 2. The two vessel occlusion model used in the study was a modification of the model described in the literature, whereby we have obviated the need to use a muscle relaxant and intubate the trachea to provide ventilatory support by providing a tight fitting face mask attached to the ventilator. Furthermore, the need to combine exsanguination and additional pharmacological means of inducing the mandatory hypotension (50 mmHg), required to decrease brain blood perfusion pressure, has been removed by simply manipulating the concentration of the already present halothane anaesthetic. 3. The appropriate level of hypotension having been reached, microvascular clips were applied to bilaterally occlude the common carotid arteries for 12 min. This resulted in a loss of the cortical EEG activity. Local cerebral blood flow was measured 6 min into the occlusion period, using the fully quantitative [14C]-iodoantipyrine autoradiographic technique, in a separate group of rats (n = 5). This illustrated the lack of any blood flow, in the areas under study, during the period when there was an isoelectric cortical EEG pattern. 4. The high grade global ischaemic lesion which occurred gave quantifiable neuronal damage in several vulnerable regions of the brain, namely, the hippocampal CA1 sub-field, cortex, thalamus, striatum, and cerebellar brain stem (Purkinje cells). 5. Following the global ischaemic insult the rats were allowed to recover for 72 h before assessment of the damage, during which time one group of rats (n = 11) received 100 micrograms kg-1 lifarizine i.a. 5 min post-occlusion, 500 micrograms kg-1 lifarizine i.p. 15 min post-occlusion, and 500 micrograms kg-1 lifarizine i.p. twice daily for 72 h. A second group of rats (n = 12) was treated with appropriate volumes of vehicle (0.4 ml kg-1 i.a. and 2 ml kg-1 i.p.) at identical time points. 6. Histopathological damage was assessed, from cresyl violet and haematoxyline/eosin stained sections, using a scoring system of 0-6 (no damage-complete neuronal death). The dosing regimen of lifarizine gave reduced damage in the hippocampal CA1 sub-field (4.1 +/- 0.3 to 2.8 +/- 0.6) and striatum (1.7 +/- 0.3 to 1.2 +/- 0.3) and significant neuroprotection in the anterior cortex (2.0 +/- 0.2 to 1.2 +/- 0.2; p < 0.05), thalamus (1.5 +/- 0.2 to 0.8 +/- 0.2; p < 0.01), posterior cortex (1.5 +/- 0.2 to 1.0 +/- 0.2; p < 0.05) and cerebellar brain stem (0.9 +/- 0.2 to 0.4 +/- 0.1; p < 0.01). The overall mean brain score was significantly reduced (from 1.5 +/- 0.1 to 0.9 +/- 0.2). 7. These data show that the newly modified 2 vessel occlusion model produced a quantifiable level of ischaemic damage and that the novel agent lifarizine is neuroprotective in the model. PMID- 8719783 TI - Nitric oxide synthase in human placenta and umbilical cord from normal, intrauterine growth-retarded and pre-eclamptic pregnancies. AB - 1. It has been suggested that a deficiency of nitric oxide (NO) may explain many of the pathophysiological features of pre-eclampsia (PE) and intra-uterine (foetal) growth retardation (IUGR). To elucidate further the role of NO in the pathophysiology of pregnancy we have determined the relative amount and activity of NO synthase (NOS) in first trimester and normal-term placental tissues, as well as in the placenta and umbilical cord in pregnancies complicated by PE and IUGR, using NG-nitro-L-[2,3,4,5(-3)H]-arginine ([3H]-L-NOARG) binding, quantitative in vitro autoradiography, [3H]-arginine to [3H]-citrulline conversion and Western blotting. 2. Specific, high affinity (KD = 38 nM) [3H]-L NOARG binding was demonstrated in the villous trophoblast of normal-term placentae. Binding was calcium-independent, stereoselective and exhibited a rank order of inhibition by NOS inhibitors and substrate (L-NOARG > or = L-NMMA > or = 7-NI > L-NAME > L-Arg > or = L-NIO > ADMA). 3. [3H]-L-NOARG binding density and NOS activity were both significantly greater in placental tissues from first trimester and PE or IUGR complicated pregnancies compared to normal-term placentae. 4. Western blotting, using an endothelial NOS peptide antiserum, demonstrated a approximately 140 KDa protein band in placental extracts and indicated that the amount of immunoreactive material was significantly greater in first trimester compared to normal-term placentae. 5. Specific [3H]-L-NOARG binding was also localized to the endothelial lining of umbilical arteries and veins, binding density being greater in the artery than the vein. [3H]-L-NOARG binding to the umbilical artery endothelium was significantly lower in PE and IUGR complicated pregnancies compared to normal-term controls. 6. The role of trophoblast-derived NO in human placental pathophysiology remains to be established, but differences in the amount of placental [3H]-L-NOARG binding, NOS activity and immunoreactive material indicate that expression of NOS in the villous trophoblast falls during pregnancy. Conversely, the apparent reduction in NOS in the umbilical artery endothelium in PE and IUGR complicated pregnancies may be indicative of endothelial dysfunction. PMID- 8719784 TI - Inhibition of cyclic GMP-dependent protein kinase-mediated effects by (Rp)-8 bromo-PET-cyclic GMPS. AB - 1. The modulation of the guanosine 3':5'-cyclic monophosphate (cyclic GMP)- and adenosine 3':5'-cyclic monophosphate (cyclic AMP)-dependent protein kinase activities by the diastereomers of 8-bromo-beta phenyl-1, N2-ethenoguanosine 3':5'-cyclic monophosphorothioate, ((Rp)- and (Sp)-8-bromo-PET-cyclic GMPS) was investigated by use of purified protein kinases. In addition, the effects of (Rp) 8-bromo-PET-cyclic GMPS on protein phosphorylation in intact human platelets and on [3H]-noradrenaline release and neurogenic vasoconstriction in electrical field stimulated rat tail arteries were also studied. 2. Kinetic analysis with purified cyclic GMP-dependent protein kinase (PKG) type I alpha and I beta, which are expressed in the rat tail artery, revealed that (Rp)-8-bromo-PET-cyclic GMPS is a competitive inhibitor with an apparent Ki of 0.03 microM. The activation of purified cyclic AMP-dependent protein kinase (PKA) type II was antagonized with an apparent Ki of 10 microM. 3. In human platelets, (Rp)-8-bromo-PET-cyclic GMPS (0.1 mM) antagonized the activation of the PKG by the selective activator 8-(4 chlorophenylthio)-guanosine 3':5'-cyclic monophosphate (8-pCPT-cyclic GMP; 0.2 mM) without affecting the activation of PKA by (Sp)-5, 6-dichloro-1-beta-D ribofurano-sylbenzimidazole- 3':5'-cyclic monophosphorothioate ((Sp)-5,6-DCl cyclic BiMPS; 0.1 mM). 4. (Rp)-8-bromo-PET-cyclic GMPS was not hydrolysed by the cyclic GMP specific phosphodiesterase (PDE) type V from bovine aorta but potently inhibited this PDE. 5. The corresponding sulphur free cyclic nucleotide of the two studied phosphorothioate derivatives, 8-bromo-beta-phenyl-1, N2 ethenoguanosine-3':5'-cyclic monophosphate (8-bromo-PET-cyclic GMP), had no effect on electrically-induced [3H]-noradrenaline release but concentration dependently decreased the stimulation-induced vasoconstriction. (Rp)-8-bromo-PET cyclic GMPS (3 microM) shifted the vasoconstriction response to the right without affecting stimulation evoked tritium overflow. 6. The NO donor, 3 morpholinosydnonimine (SIN-1) relaxed rat tail arteries precontracted with phenylephrine (1 microM). The SIN-1 concentration-relaxation curve was shifted in a parallel manner to the right by (Rp)-8-bromo-PET-cyclic GMPS, suggesting that the relaxation was mediated by a cyclic GMP/PKG-dependent mechanism. 7. The [3H] noradrenaline release-enhancing effect and stimulation-induced decrease in vasoconstriction of forskolin were unaffected by (Rp)-8-bromo-PET-cyclic GMPS. Moreover, the forskolin concentration-relaxation curve was not changed in the presence of the PKG inhibitor, suggesting a high selectivity in intact cells for PKG- over PKA-mediated effects. 8. The results obtained indicate that (Rp)-8 bromo-PET-cyclic GMPS presently is the most potent and selective inhibitor of PKG and is helpful in distinguishing between cyclic GMP and cyclic AMP messenger pathways activation. Therefore, this phosphorothioate stereomer may be a useful tool for studying the role of cyclic GMP in vitro. PMID- 8719785 TI - Effect of endopeptidase-24.11 inhibitors and C-ANP receptor ligand on responses evoked in arterioles of rat cremaster muscle by atrial natriuretic peptide. AB - 1. The present study examined the effect of exogenous atrial natriuretric peptide (ANP), alone or in presence of inhibitors of the two major mechanisms for clearing ANP, metabolism by neutral endopeptidase-24.11 (NEP) and internalization by C-ANP receptors, on arteriolar responses using intravital microscopy on the rat cremaster muscle after intravenous or topical administration of the peptide. 2. Topical application of ANP (3 x 10(-10) to 3 x 10(-8) M) produced a gradual increase in arteriolar diameter. NEP inhibitors, thiorphan (30 mg kg-1, i.v.), kelatorphan (10 mg kg-1, i.v.) and retrothiorphan (25 mg kg-1, i.v.) alone, did not significantly affect vascular tone but caused significant potentiation of the arteriolar responses to topically applied ANP. 3. When given as an i.v. bolus, ANP dilates skeletal arterioles at a high dose (20 micrograms kg-1). At a lower dose (10 micrograms kg-2), ANP alone or with retrothiorphan or the C-ANP receptor ligand C-ANP (4-23) did not produce any arteriolar responses, while after the combined administration of the two inhibitors, an increase in arteriolar diameter was induced. 4. These results indicate that low doses of topically applied ANP dilate rat cremaster arterioles and that the vasodilator responses can be potentiated by NEP inhibition. When given as an i.v. bolus, a high dose of ANP can also dilate skeletal arterioles. However at a lower dose the rapid metabolism of the peptide prevents it from producing its action. PMID- 8719786 TI - Changes in [3H]-UK 14304 binding to alpha 2-adrenoceptors in morphine-dependent guinea-pigs. AB - 1. The aim of this study was to investigate the effect of a noradrenergic input in the cortex of morphine-dependent animals. Binding of the alpha 1-adrenoceptor ligand [3H]-prazosin did not change in cortical membranes taken from morphine dependent as compared to control guinea-pigs. However, binding of the alpha 2 adrenoceptor ligand [3H]-UK 14304 showed decreased KD (-30%) in the absence of significant changes in Bmax, either in cortical membranes or in synaptosomes. 2. Several characteristics of this phenomenon were identified. First, it occurs in a time-dependent fashion, in that it takes 5 days of chronic morphine treatment to start developing. Second, it can be observed after acute administration of high doses of morphine (100 mg kg-1). Third, it does not require a connection with the locus coeruleus or with other subcortical structures, in that it can be reproduced in vitro in isolated cortical slices. Fourth, it requires the integrity of cortical structures, since it cannot be reproduced in vitro in cortical synaptosomes. 3. Release studies were run to attempt identification of a functional correlate of the above observations. No changes were observed in the ability of the alpha 2-adrenoceptor agonist UK 14304 to inhibit 35 mM K(+)-evoked [3H]-noradrenaline outflow from cortical synaptosomes taken from morphine dependent as compared to control guinea-pigs. However, a large decrease in the IC50 of UK 14304 for the inhibition of 35 mM K(+)-evoked [3H]-gamma-aminobutyric acid ([3H]-GABA) outflow (41 vs. 501 nM) was observed in morphine-dependent as compared to control animals. 4. These data suggest that, in the guinea-pig, chronic morphine treatment is associated with a shift from a low to high affinity agonist state in alpha 2-adrenoceptors on cortical GABA terminals. PMID- 8719787 TI - Meta-chlorophenylpiperazine attenuates formalin-induced nociceptive responses through 5-HT1/2 receptors in both normal and diabetic mice. AB - 1. This study was designed to investigate the effect of meta chlorophenylpiperazine (m-CPP; a 5-hydroxytryptamine (5-HT) receptor agonist) on the formalin-induced nociceptive responses in normal, insulin-dependent streptozotocin (STZ) diabetic and non-insulin dependent genetically diabetic (db/db) mice. 2. A subcutaneous injection of diluted formalin (1% formaldehyde in 0.9% saline, 10 microliters) under the plantar surface of the left hindpaw induced biphasic nociceptive responses, the first and second phases considered to represent acute and chronic pain, respectively. The former response in db/db mice was significantly lower than those in normal mice, and the latter responses in STZ and db/db mice were significantly lower than those in normal mice. 3. In normal mice, m-CPP (0.32-3.2 mg ml-1, p.o.) exhibited potent antinociceptive activity, dose-dependently attenuating the first and second phase; the ID50 value of the second phase was 0.4 mg kg-1. m-CPP (0.32-3.2 mg kg-1, p.o.) also dose dependently attenuated the formalin-induced nociceptive responses in STZ-induced diabetic mice and genetically diabetic db/db mice, and the activities were comparable to those in normal mice. 4. The antinociceptive activities of m-CPP (1 mg kg-1, p.o.) were significantly inhibited by pretreatment with pindolol (a 5 HT1-receptor antagonist, 1 mg kg-1, i.p.) or ketanserin (a 5-HT2 receptor antagonist, 1 mg kg-1, i.p.) but were hardly affected by ICS205-930 (a 5-HT3 receptor antagonist, 1 mg kg-1, i.p.). 5. These results suggest that m-CPP inhibits not only acute but also chronic pain transmission through 5-HT1 and 5 HT2 receptors, and that the 5-hydroxytryptaminergic antinociceptive pathways are little affected by diabetes. PMID- 8719788 TI - The pharmacological properties of K+ currents from rabbit isolated aortic smooth muscle cells. AB - 1. Using the whole-cell patch-clamp technique, the effects of several K+ channel blocking drugs on K+ current recorded from rabbit isolated aortic smooth muscle cells were investigated. 2. Upon depolarization from -80 mV, outward K+ current composed of several distinct components were observed: a transient, 4 aminopyridine (4-AP)-sensitive component (I1) and a sustained component (Isus), comprising a 4-AP-sensitive delayed rectifier current (IK(V)), and a noisy current which was sensitive to tetraethylammonium (TEA), and probably due to Ca(2+)-activated K+ current (IK(Ca)). 3. Several drugs in clinical or experimental use have as part of their action an inhibitory effect on specific K+ channels. Because of their differential K+ channel blocking effects, these drugs were used in an attempt to characterize further the K+ channels in rabbit aortic smooth muscle cells. Imipramine, phencyclidine, sotalol and amitriptyline failed to block selectively any of the components of K+ current, and were thus of little value in isolating individual channel contributions. Clofilium showed selective block of IK(V) in the presence of TEA, but only at low stimulation frequencies (0.07 Hz). At higher frequencies (1 Hz) of depolarization, both I1 and IK(V) were suppressed to a similar extent. Thus, the blocking action of clofilium was use dependent. 4. The voltage-dependent inactivation of I1 and the delayed rectifier were very similar although a brief (100 ms) pre-pulse to -30 mV could preferentially inactivate I1. Together with the non-selective blocking effects of the K+ channel blockers, similarities in the activation and inactivation of these two components suggest that they may not exist as separate ionic channels, but as distinct kinetic states within the same K+ channel population. 5. The effects of all of these drugs on tension were examined in strips of rabbit aorta. The non specific K+ channel blockers caused only minor increases in basal tension. TEA and 4-AP by themselves caused significant increases in tension and were even more effective when applied together. There appeared to be no correlation between the effects of the drugs tested on tension and their actions on currents recorded from isolated myocytes. Thus tension studies are an inappropriate means of investigating the mechanism of action of these drugs, and studies on ionic currents in isolated myocytes cannot easily predict drug actions on intact tissues. PMID- 8719789 TI - The pharmacology of GR203040, a novel, potent and selective non-peptide tachykinin NK1 receptor antagonist. AB - 1. The in vitro and in vivo pharmacology of GR203040 ((2S, 3S)-2-methoxy-5 tetrazol-1-yl-benzyl-(2-phenyl-piperidin-3-y l)-amine), a novel, highly potent and selective non-peptide tachykinin NK1 receptor antagonist, was investigated in the present study. 2. GR203040 potently inhibited [3H]-substance P binding to human NK1 receptors expressed in Chinese hamster ovary (CHO) and U373 MG astrocytoma cells, and NK1 receptors in ferret and gerbil cortex (pKi values of 10.3, 10.5, 10.1 and 10.1 respectively). GR203040 had lower affinity at rat NK1 receptors (pKi = 8.6) and little affinity for human NK2 receptors (pKi < 5.0) in CHO cells and NK3 receptors in guinea-pig cortex (pKi < 6.0). With the exception of the histamine H1 receptor (pIC50 = 7.5). GR203040 had little affinity (pIC50 < 6.0) at all non-NK1 receptors and ion channels examined. Furthermore, GR203040 produced only weak inhibition of Na+ currents in SH-SY5Y neuroblastoma and superior cervical ganglion cells (pIC50 values < 4.0). GR203040 produced only weak antagonism of Ca(2+)-evoked contractions of rat isolated portal vein (pKn = 4.1). The enantiomer of GR203040, GR205608 (2R, 3R)-2-methoxy-5-tetrazol-1-yl benzyl-(2-phenyl-piperidin-3-y l)-amine), had 10,000 fold lower affinity at the human NK1 receptor expressed in CHO cells (pKi = 6.3). 3. In gerbil ex vivo binding experiments, GR203040 produced a dose-dependent inhibition of the binding of [3H]-substance P to cerebral cortical membranes (ED50 = 15 micrograms kg-1 s.c. and 0.42 mg kg-1 p.o.). At 10 micrograms kg-1 s.c., the inhibition of [3H] substance P binding was maintained for > 6 h. In the rat, GR203040 was less potent (ED50 = 15.4 mg kg-1 s.c.) probably reflecting, at least in part, its lower affinity at the rat NK1 receptor. 4. In guinea-pig isolated ileum and dog isolated middle cerebral and basilar arteries, GR203040 produced a rightward displacement of the concentration-effect curves to substance P methyl ester (SPOMe) with suppression of the maximum agonist response (apparent pKB values of 11.9, 11.2 and 11.1 respectively). 5. In anaesthetized rabbits, GR203040 antagonized reductions in carotid arterial vascular resistance evoked by SPOMe, injected via the lingual artery (DR10 (i.e. the dose producing a dose-ratio of 10) = 1.1 micrograms kg-1, i.v.). At a dose 20 fold greater than its DR10 value (i.e. 22 micrograms kg-1, i.v.), significant antagonism was evident more than 2 h after GR203040 administration. 6. In anaesthetized rats, GR203040 (3 and 10 mg kg 1, i.v.) produced a dose-dependent inhibition of plasma protein extravasation in dura mater, conjunctiva, eyelid and lip in response to electrical stimulation of the trigeminal ganglion. 7. It is concluded that GR203040 is one of the most potent and selective NK1 receptor antagonists yet described, and as such, has considerable potential as a pharmacological tool to characterize the physiological and pathological roles of substance P and NK1 receptors. GR203040 may also have potential as a novel therapeutic agent for the treatment of conditions such as migraine, emesis and pain. PMID- 8719790 TI - The broad-spectrum anti-emetic activity of the novel non-peptide tachykinin NK1 receptor antagonist GR203040. AB - 1. Following our earlier observations that the tachykinin NK1 receptor antagonist CP-99,994 is an effective anti-emetic in ferrets, we have examined the anti emetic effects of a more potent and novel NK1 receptor antagonist, GR203040, against various emetic stimuli in the ferret, dog and house musk shrew (Suncus murinus). 2. In ferrets, GR203040 (0.1 mg kg-1 s.c. or i.v.) is effective against emesis induced by radiation, cisplatin, cyclophosphamide, copper sulphate, ipecacuanha or morphine. 3. In animals in which emesis had been established with cisplatin, GR203040 (1 mg kg-1 s.c.) was fully effective as an interventional treatment. No further emesis was seen in animals treated with GR203040 whilst saline-treated animals continued to vomit. 4. GR203040 (0.1 mg kg-1 s.c.) retains anti-emetic efficacy in the ferret, even when given as a 6 h pretreatment, indicating that this compound has a long duration of action. The compound is also effective orally at the same dose, when given as a 90 min pretreatment. 5. GR203040 (0.1 mg kg-1 i.v.) is fully effective against ipecacuanha-induced emesis in the dog. 6. GR203040 is effective against motion- and cisplatin-induced emesis in Suncus murinus. These effects were seen at doses an order of magnitude greater than those shown to be effective against cisplatin in the ferret. 7. In conclusion, GR203040 is a novel anti-emetic agent, and the broad spectrum of anti emetic activity, together with activity observed in three species, suggests that this compound is worthy of clinical investigation. PMID- 8719791 TI - Inhibition of bradykinin-evoked trigeminal nerve stimulation by the non-peptide bradykinin B2 receptor antagonist WIN 64338 in vivo and in vitro. AB - 1. This study investigated the effect of the recently described non-peptide bradykinin B2 receptor antagonist, WIN 64338 ([[4-[[2- [[bis(cyclohexylamino)methylene]amino]-3-(2-naphthalenyl)-1-oxopropyl] amino]phenyl]methyl]tributylphosphoniumchloride monohydrochloride), in experimental models of bradykinin-evoked sensory nerve stimulation. 2. In the rabbit isolated iris sphincter in vitro, bradykinin-evoked contractile responses are mediated via tachykinins released from peripheral endings of the trigeminal sensory nerve. WIN 64338 (1-10 microM) competitively antagonised contractile responses to bradykinin with a pKB estimate of 6.6 +/- 0.1 (n = 11). The antagonism was selective since WIN 64338 (10 microM) did not significantly inhibit submaximal contractile responses to the direct-acting spasmogens substance P (10 nM), neurokinin A (3 nM), substance P methyl ester (10 nM) or senktide (100 nM); nor by sensory non-adrenergic non-cholinergic nerve stimulation evoked by capsaicin (10 microM), or electrical field-stimulation (3, 10, 30 Hz) (P > 0.05; n = 3-11). 3. Topical application of bradykinin to the conjunctiva and to the nasal mucosa of the guinea-pig in vivo causes plasma extravasation predominantly via the release of tachykinins from peripheral endings of the trigeminal nerve. The increases in plasma extravasation (measured by extravasation of Evans blue dye) induced by bradykinin in the guinea-pig conjunctiva (20 nmol) and nasal mucosa (50 nmol) were markedly reduced (by 81 +/- 3% and 69 +/- 5%, respectively) following pretreatment with WIN 64338 (30 nmol kg 1, i.v.) (n = 5-6; P < 0.05), with almost complete inhibition at a higher dose of WIN 64338 (300 nmol kg-1, i.v.; n = 5-6). This inhibition was selective since at 300 nmol kg-1, WIN 64338 did not inhibit plasma extravasation evoked by substance P in the conjunctiva (5 nmol; P > 0.05; n = 6) or in the nasal mucosa (50 nmol; P > 0.05; n = 5). 4. This study demonstrates that WIN 64338 is a selective and competitive bradykinin B2 receptor antagonist and can be useful for analysing bradykinin-evoked trigeminal nerve stimulation both in vitro and in vivo. PMID- 8719792 TI - Differential effect of phosphodiesterase 4 inhibitors on the proliferation of human peripheral blood mononuclear cells from normals and subjects with atopic dermatitis. AB - 1. The aims of this study were to compare the effects of selective inhibitors of the type 3, type 4 and type 5 phosphodiesterase (PDE) isoenzymes on the phytohaemagglutinin (PHA)-stimulated proliferation of human peripheral blood mononuclear cells (HPBM) from normals and subjects with atopic dermatitis (AD). 2. Mononuclear cells were isolated from peripheral venous blood of normals and subjects with AD. A concentration-response curve was carried out with PHA (0.5-5 micrograms ml-1) and a concentration which produced a submaximal stimulation of proliferation (2 micrograms ml-1) was selected for further experiments. HPBM (10(5) cells per well) were stimulated with PHA (2 micrograms ml-1) in the absence or presence of PDE inhibitor (0.01 microM-10 microM) and 24 h later [3H] thymidine (0.1 microCi per well) was added. Cells were incubated for an additional 24 h period and [3H]-thymidine incorporation measured. 3. The type 4 PDE inhibitors (rolipram, RO 20-1724 and denbufylline) produced a concentration related inhibition of proliferation of HPBM from normal and AD subjects. The IC50 for rolipram was significantly (P < 0.05) lower in HPBM from AD patients 0.28 microM (95% confidence limits (CL): 0.158-0.499, n = 5) vs normal subjects 2.6 microM (95% CL: 0.867-7.05, n = 5, P < 0.05) as were the IC50 values for denbufylline: 0.26 microM (95% CL: 0.152-0.440, n = 5) vs 1.84 microM (95% CL: 0.467-7.23, n = 5, P < 0.05) respectively and RO 20-1724: 1.49 microM (95% CL: 0.61 microM-3.64 microM) vs 6.46 microM (95% CL: 2.03 microM-20.46 microM), respectively. 4. The mixed type 3/4 inhibitors (zardaverine and benzafentrine) produced a concentration-related inhibition of proliferation of HPBM from normal and AD subjects. The IC50 value for zardaverine in HPBM from normal subjects: 1.8 microM (95% CL: 0.43 microM-7.85 microM, n = 4) was similar to that in AD subjects: 1.03 microM (95% CL: 0.48 microM-2.28 microM) as was the IC50 value for benzafentrine in normal 3.8 microM (95% CL: 2.45 microM-5.9 microM) and atopic 5.5 microM (95% CL: 3.84 microM-7.78 microM) HPBM. The type 5 PDE inhibitor, zaprinast was ineffective at inhibiting the proliferation of normal HPBM. The type 3 PDE inhibitor, siguazodan only inhibited [3H]-thymidine incorporation at a concentration of 10 microM. 5. These results show that combined inhibition of the type 3 and 4 PDE isoenzymes in HPBM from normal subjects has a greater antiproliferative effect than inhibition of the type 4 isoenzyme alone. In addition these data indicate that the proliferative response of HPBM from AD subjects is more sensitive to PDE 4 inhibition than the proliferation of HPBM from normals. PMID- 8719793 TI - Microvascular mechanisms of histamine-induced potentiation of leukocyte adhesion evoked by chemoattractants. AB - 1. Intravital microscopy of the rat mesentery was used to examine interactions between histamine and the chemoattractant leukotriene B4 (LTB4) with regard to leukocyte adhesion in postcapillary venules. 2. Topical administration of histamine caused a four fold potentiation of LTB4-induced leukocyte adhesion. 3. Histamine significantly increased the rolling leukocyte flux by 25%, and this effect of histamine on rolling was strictly blood flow-dependent, i.e. we found significant positive correlations between both blood flow and total leukocyte flux and between total and rolling leukocyte flux, while no changes in leukocyte rolling fraction or rolling velocity were observed. Furthermore, histamine caused a clear-cut increase in venular plasma protein leakage. 4. The platelet activating factor (PAF) receptor antagonist WEB 2086, which effectively inhibited adhesion of leukocytes evoked by exogenous PAF, did not reduce the potentiating effect of histamine on LTB4-induced leukocyte adhesion. 5. The vasodilator acetylcholine (ACh) caused a moderate enhancement of LTB4 induced leukocyte adhesion in proportion to its blood flow-dependent 40% increase in rolling leukocyte flux. In contrast to histamine, ACh did not provoke vascular leakage of plasma proteins. 6. Taken together, our findings suggest that histamine plays an important pro-inflammatory role in tissues where leukocyte rolling is already present, by potentiating chemoattractant-induced firm leukocyte adhesion through a combination of microcirculatory changes such as increased rolling leukocyte flux and vascular permeability. PMID- 8719794 TI - Lack of a centrally-mediated antihypertensive effect following acute or chronic central treatment with AT1-receptor antagonists in spontaneously hypertensive rats. AB - 1. The role of the central renin-angiotensin system in the pathogenesis of hypertension in spontaneously hypertensive rats (SHR) was examined following acute and chronic intracerebroventricular (i.c.v.) infusions of angiotensin1 (AT1) receptor antagonists. 2. Groups of SHR were chronically instrumented for acute i.c.v. administration of the AT1 receptor antagonists, losartan and CV 11974, on mean arterial blood pressure (MAP) and heart rate (HR). Other groups of SHR also had mini-osmotic pumps implanted for chronic i.c.v. infusion of CV 11974. 3. Initially both young (15-18 weeks, n = 8) and old (25-29 weeks, n = 9) SHR received acute i.c.v. injections of losartan (10 micrograms) while a third group of young SHR received CV-11974 (1 microgram, n = 6). In all three groups of SHR, MAP and HR did not change up to 24 h after antagonist injection. However, changes in MAP and HR in response to i.c.v. angiotensin II (AII, 100 ng) were abolished 15 min after administration of the AT1 receptor antagonists. These responses had returned to control levels after 3 h in both groups given losartan but were still significantly depressed at 24 h in the CV-11974-treated group. By contrast, responses to i.v. AII (25 ng) before and 1 h after administration of AT1 receptor antagonists were not significantly different. 4. For chronic studies, four groups of SHR received chronic i.c.v. infusion of either vehicle (n = 9) or CV-11974 (1, 5 and 100 micrograms kg-1 day-1) (n = 4, 7 and 8 respectively) for 4 days. Baseline cardiovascular parameters were monitored daily together with changes in MAP and HR in response to both i.c.v. and i.v. AII (100 ng and 50 ng respectively) and i.v. phenylephrine (3 micrograms). Responses to i.c.v. carbachol (5 micrograms) were also recorded on day 4 while baroreflex function was assessed between days 1-3. In SHR treated chronically with i.c.v. vehicle or CV-11974, at 1 or 5 micrograms kg-1 day-1, resting MAP and HR did not vary over the four day infusion period. However, SHR treated with 100 micrograms kg-1 day-1 CV-11974 had significantly lower MAP compared to vehicle-treated SHR. While there was some variation in resting HR, there were no differences between the drug-treated and vehicle-treated groups. Pressor responses following i.c.v. AII administration were slightly, but significantly, inhibited on days 3 and 4 in the low dose CV-11974-treated (1 microgram kg-1 day-1) SHR. However, these responses were abolished on all 4 days in the 5 and 100 micrograms kg-1 day-1 CV 11974-treated groups. By contrast, changes in MAP and HR following i.v. AII injection did not vary over the 4 day infusion between SHR treated with the 2 lowest doses of CV-11974 and the vehicle-treated group. However, in the high dose CV-11974-treated SHR (100 micrograms kg-1 day-1), the cardiovascular effects of AII were abolished. In addition, phenylephrine (i.v.) and carbachol (i.c.v.) induced changes in MAP and HR were not significantly different in all four treatment groups. Similarly, baroreflex function was unaffected by i.c.v. infusion of 100 micrograms kg-1 day-1 CV-11974, except for a significant fall in BP50 which paralleled the fall in resting MAP. 5. Collectively, these results indicate that acute and chronic central AT1 receptor antagonism does not lower MAP in conscious SHR in doses which only block central AII-induced pressor activity. Chronic central infusion of CV-11974 at sufficiently high doses will lower MAP, as has been reported by others, but not without the abolition of the peripheral effects of AII. Therefore it is most likely that peripheral AT1 receptor blockade contributes to the hypotensive action of CV-11974 under these conditions. PMID- 8719795 TI - Role for intracellular platelet-activating factor in the circulatory failure in a model of gram-positive shock. AB - 1. This study investigates the effects of two structurally different antagonists of platelet-activating factor (PAF), BN52021 and WEB2086, on the circulatory and renal failure elicited by lipoteichoic acid (LTA) from Staphylococcus aureus (an organism without endotoxin) in anaesthetized rats. 2. Administration of LTA (10 mg kg-1, i.v.) caused hypotension and vascular hyporeactivity to noradrenaline (1 microgram kg-1, i.v.) WEB2086 (5 mg kg-1, i.v., 20 min before and 150 min after LTA) inhibited the delayed fall in mean arterial blood pressure (at 300 min: 99 +/- 6 mmHg vs. 75 +/- 6 mmHg, P < 0.01) and prevented the decrease in pressor response to noradrenaline (at 300 min: 36 +/- 5 mmHg min vs. 17 +/- 5 mmHg min, P < 0.01). Surprisingly, BN52021 (20 mg kg-1, i.v., 20 min before and 150 min after LTA) neither prevented the hypotension (74 +/- 6 mmHg) nor the vascular hyporeactivity (21 +/- 5 mmHg min). However, BN52021 inhibited the hypotension to injections of PAF as well as the circulatory failure elicited by lipopolysaccharides (10 mg kg-1, i.v.). 3. LTA caused an increase in plasma concentration of creatinine from 39 +/- 5 microM (sham-operated) to 70 +/- 8 microM and urea from 4.7 +/- 0.1 to 13.1 +/- 1.6 mM. The renal failure elicited by LTA was significantly inhibited by WEB2086 (creatinine: 45 +/- 4 microM and urea: 5.7 +/- 0.7 mM), but not by BN52021. 4. The induction of nitric oxide synthase activity in lungs by LTA was attenuated by WEB2086 from 98 +/- 17 to 40 +/- 15 pmol L-citrulline 30 min-1 mg-1 protein (P < 0.01), but not by BN52021 (148 +/- 21 pmol L-citrulline 30 min-1 mg-1 protein). Similarly, WEB2086, but not BN52021, inhibited the increase in plasma nitrite concentration associated with the delayed circulatory failure caused by LTA. The release of tumour necrosis factor-alpha (TNF-alpha) after injection of LTA was not attenuated by WEB2086. 5. The induction of nitrite release by cultured macrophages activated with LTA (10 micrograms ml-1 for 24 h) was inhibited by 74 +/- 4% by WEB2086 (3 x 10(-4) M), but not by BN52021, indicating that only WEB2086 acts on intracellular PAF receptors. 6. Thus, the intracellular release of PAF contributes to the circulatory and renal failure and induction of nitric oxide synthase elicited by LTA in anaesthetized rats. The difference between the two structurally different PAF antagonists in our septic shock models using either LTA or lipopolysaccharide (LPS), shows the importance of models for Gram-positive sepsis in the elucidation of the pathophysiology of septic shock and for the evaluation of potential drugs. PMID- 8719797 TI - Relationship between morphine analgesia and cortical extracellular fluid levels of morphine and its metabolites in the rat: a microdialysis study. AB - 1. The effect of morphine (10 mg kg-1, s.c.) on the analgesic response measured by the tail-flick method was determined in male Sprague-Dawley rats. The analgesic response to morphine was correlated with the levels of morphine and its metabolites collected by microdialysis from the cortical extracellular fluid (ECF). 2. The analgesic response to morphine lasted for 4 h. The concentration of morphine during a 4 h collection period was significantly higher than the metabolites concentration. The relative concentration of morphine and its metabolites during the 4 h period was 70 and 30% respectively. 3. The analgesic response during the first 2.25 h period accounted for more than 82% of the total analgesia as determined by the area under the time-response curve (AUC). The concentration of morphine and its metabolites during the same period were 78 and 22%, respectively, but they did not differ during the 2.25-4.0 h period (52 and 48%). 4. The half-life for morphine and its metabolites were similar, the maximal achievable concentration Cmax and AUC0-4 h were lower for metabolites but the time to reach maximum concentration was higher for morphine metabolites than for morphine. The ratio of the concentration of metabolites to the concentration of morphine in the cortical ECF increased with time whereas the analgesic response to morphine decreased with time. 5. At several time points following morphine injection even though the levels of morphine were the same, the concentration of metabolites (mainly M3G) differed and thus the ratio [metabolite/morphine]. A plot of [metabolite]/[morphine] vs. analgesia gave a high correlation coefficient. Since M3G has been shown to be antianalgesic and is the only metabolite of morphine in the rat, it is concluded that the levels of this metabolite may regulate the analgesic effect of morphine in the rat. PMID- 8719796 TI - Attenuation by valproate of c-fos immunoreactivity in trigeminal nucleus caudalis induced by intracisternal capsaicin. AB - 1. Valproic acid, useful in the treatment of migraine, is an inhibitor of gamma aminobutyric acid (GABA) aminotransferase and activator of glutamic acid decarboxylase. Its mechanism in migraine remains obscure. The effects of valproic acid (2-propylpentanoic acid) were examined on the number of cells expressing c fos-like immunoreactivity (c-fos-LI), a marker of neuronal activation, within the trigeminal nucleus caudalis (lamina I, IIo, TNC) 2 h after intracisternal injection of the irritant, capsaicin (0.1 ml; 15.25 micrograms ml-1), in urethane anaesthetized Hartley guinea-pigs. Positive cells were counted in eighteen sections (50 microns) at three representative levels (rostral, middle and caudal) within lamina I, IIo of the TNC in 90 animals. 2. Numerous cells were labelled after capsaicin instillation (244 +/- 25; 1 ml; 15.25 mM) but not after capsaicin vehicle (11 +/- 1). Positive cells were also found within the medial reticular nucleus, the area postrema and the nucleus of the solitary tract. A similar distribution has been demonstrated previously after application of intracisternal irritants such as autologous blood or carrageenin. 3. Valproate (> or = 10 mg kg 1, i.p.) reduced labelled cells by 52% (P < 0.05) in lamina I, IIo but not within the area postrema, the nucleus of the solitary tract or the medial reticular nucleus. A similar finding was obtained previously after administration of sumatriptan, dihydroergotamine or the NK1 receptor antagonist RPR 100,893. 4. Pretreatment with bicuculline (30 micrograms kg-1; i.p.), a GABAA antagonist, but not phaclofen (1 mg kg-1) a GABAB antagonist, reversed the effect of valproate and increased c-fos positive cells within lamina I, IIo. Somewhat paradoxically, bicuculline by itself (30 micrograms kg-1 i.p.) decreased the number of labelled cells suggesting that more than a single GABAergic mechanism can suppress c-fos expression. 5. We conclude that the mechanism of action of valproate is mediated via GABAA receptors. Since valproate decreases both c-fos expression and as previously shown, neurogenic inflammation within the meninges, the GABAA receptor complex might provide an important target for drug development in migraine and related headaches. PMID- 8719798 TI - Electrophysiological basis for antiarrhythmic efficacy, positive inotropy and low proarrhythmic potential of (-)-caryachine. AB - 1. (-)-Caryachine, isolated from the plant (Cryptocarya chinensis), increased the contractility of atrial and right ventricular strips and significantly suppressed the reperfusion arrhythmias in adult rabbit heart (ED50 = 1.27 microM). 2. Data obtained by the whole-cell voltage clamp technique has shown that (-)-caryachine causes a negative shift of the steady-state Na channel inactivation and a slower rate of recovery from inactivation. The maximal Na current amplitude decreased to 67 +/- 7%, 29 +/- 8% and 12 +/- 5% after 0.5, 1.5 and 4.5 microM (-)-caryachine, respectively. 3. This agent also had effects on the time- and voltage-dependent K currents. (-)-Caryachine markedly suppressed the 4-AP-sensitive transient outward current (I10). However, it produced very little voltage-dependent shift in inactivation. After 0.5, 1.5 and 4.5 microM of the compound, the respective value of I10 elicited at +60 mV was 80 +/- 7%, 45 +/- 8% and 15 +/- 3%. At higher concentrations, the inward rectifier K current (IK1) was also inhibited but to a much smaller extent. Its slope conductance after 0.5, 1.5 and 4.5 microM (-) caryachine was reduced to 71 +/- 9%, 51 +/- 12% and 42 +/- 11%, respectively. The outward hump of inward rectification was not changed. 4. In contrast, the L-type Ca current was not significantly changed by (-)-caryachine. 5. Electrophysiological studies in perfused whole heart preparations revealed that ( )-caryachine increased the intra-atrial conduction interval and also prolonged the atrial refractory period. No proarrhythmic effects were induced during the infusion of this compound (up to 13.5 microM). 6. We conclude that (-)-caryachine predominantly blocks the Na and I10 currents. These changes alter the electrophysiological properties of the heart and terminate the induced ventricular arrhythmias. The relatively selective I10 inhibition, safety margin of Ik1 suppression and lack of effect on Ica-L will provide an opportunity to develop an effective antiarrhythmic agent with positive inotropy as well as low proarrhythmic potential. PMID- 8719799 TI - The effect of glucocorticoids on proliferation of human cultured airway smooth muscle. AB - 1. Airway smooth muscle proliferation is a significant component of the airway wall remodelling that occurs in asthma. In this study, the effects of glucocorticoids on mitogenic responses of human airway smooth muscle have been examined. 2. Pretreatment of smooth muscle cells with dexamethasone (100 nM, 60 min) inhibited thrombin-induced increases in [3H]-thymidine incorporation (DNA synthesis) and cell number. 3. Inhibition of thrombin-induced [3H]-thymidine incorporation was also observed with hydrocortisone (0.01-1 microM) and methylprednisolone (0.001-0.1 microM) pretreatment. In contrast, pretreatment with either testosterone (0.001-1 microM) progesterone (0.001-1 microM), 17 beta oestradiol (0.001-1 microM), or aldosterone (0.001-1 microM) had no effect on the response to thrombin. 4. Responses to a range of mitogens including thrombin (0.01-. 10 u ml-1), epidermal growth factor (EGF, 3-3000 pM), basic fibroblast growth factor (bFGF, 0.3-300 pM) and foetal calf serum (FCS, 0.1-10% v/v) were inhibited by dexamethasone (100 nM) pretreatment. However, the magnitude of the inhibitory effect was dependent on the mitogen, with EGF being the least, and thrombin being the most sensitive to the inhibitory effect. 5. The potency of hydrocortisone as an inhibitor of [3H]-thymidine incorporation was reduced when FCS (10% v/v, which caused a 40 fold increase in [3H]-thymidine incorporation) was used as the mitogen in place of thrombin (0.3 u ml-1, which caused a 10 fold increase in [3H]-thymidine incorporation). 6. The effect of post-treatment with dexamethasone (100 nM) indicated that addition of the glucocorticoid up to 17-19 h after thrombin (0.3 u ml-1) produced similar degrees of inhibition to those obtained when it was added as a pretreatment. Dexamethasone no longer produced an inhibitory effect if added 21 h or more after the addition of thrombin. 7. These results suggest that glucocorticoids regulate airway smooth muscle proliferation initiated by a range of stimuli. This effect may be of importance in the therapeutic actions of these compounds in asthma, particularly when they are used for prolonged periods of time. PMID- 8719800 TI - Inhibition by riluzole of glycinergic postsynaptic currents in rat hypoglossal motoneurones. AB - 1. Riluzole has been shown to have beneficial effects in motoneurone disease, yet its effect on motoneurones is not known. To address this question, we investigated synaptic modulation by riluzole in hypoglossal motoneurones by recording glycinergic inhibitory postsynaptic currents evoked by stimulation of nearby single interneurones. 2. Glycinergic inhibitory postsynaptic currents were evoked by electrical stimulation of single interneurones and were recorded from visually identified hypoglossal motoneurones. Riluzole (10 microM) inhibited mean amplitude of evoked glycinergic inhibitory postsynaptic currents by 87%. 3. We found that riluzole suppressed sodium currents in brainstem interneurones by 23.8%. Riluzole did not modulate barium currents through voltage-activated calcium channels (98% of control). Therefore, the effect of riluzole on synaptic transmission may be mediated, in part, by stabilizing presynaptic neurones through inhibition of voltage-activated sodium currents. 4. In the presence of tetrodotoxin (0.5 microM), riluzole reduced the frequency (1.2 Hz in control to 0.6 Hz in riluzole) of spontaneous transmitter release recorded in motoneurones. 5. Riluzole was found to have no effect on mean miniature inhibitory postsynaptic current amplitude, therefore the reduction in spontaneous transmitter release cannot be due to an action on postsynaptic glycine receptors. 6. We conclude that riluzole inhibits synaptic transmission presynaptically, independent of a reduction in the excitation of presynaptic neurones. PMID- 8719801 TI - Inhibition of relaxations to nitrergic stimulation of the mouse anococcygeus by duroquinone. AB - 1. The role of copper/zinc superoxide dismutase (Cu/Zn SOD) in protection of nitrergic neurotransmission in the mouse anococcygeus was investigated by use of duroquinone (DQ), which generates superoxide anions within tissues via reduction by flavoprotein enzymes. 2. In control anococcygeus muscles, DQ (10-100 microM) produced concentration-related inhibition (-log IC40 = 4.41) of relaxations to exogenous nitric oxide (NO; 15 microM). Nitrergic relaxations induced by field stimulation (10 Hz; 10 s train) were much less affected, 100 microM DQ reducing nitrergic relaxations by only 14 +/- 6%. 3. Following incubation with the Cu/Zn SOD inhibitor, diethyldithiocarbamate (DETCA; 3 mM; 45 min incubation; 10 min washout), the inhibitory effects of DQ on relaxations to NO were potentiated ( log IC40 = 5.22), and clear, concentration-related inhibitions of nitrergic relaxations were now observed (-log IC40 = 4.54). In both cases, these inhibitions were partially reversed by Cu/Zn SOD (250 u ml-1). In DETCA-treated tissues, DQ (100 microM) also reduced relaxations to sodium nitroprusside (1 microM) and S-nitroso-glutathione (30 microM), but potentiated those to 8-Br cyclic GMP (100 microM). 4. Neither hydroquinone (HQ: 100 microM) nor 1,4 benzoquinone (BQ: 100 microM), both of which reduced responses to exogenous NO, inhibited relaxations induced by field stimulation in DETCA-treated tissues. Indeed, when added during DQ-induced inhibition of nitrergic relaxations, both HQ and BQ produced partial reversal of the block. 5. DQ had no effect on the detection of superoxide anions estimated via the xanthine:xanthine oxidase chemiluminescence assay, or of authentic NO as measured by a chemical microsensor. However, the detection of both superoxide anions and NO in these assays was inhibited by inclusion of either HQ or BQ. 6. The results support the proposal that nitrergic transmission in the peripheral nervous system is protected by Cu/Zn SOD activity in the region of the neuroeffector junction, and this may explain the lack of effect of superoxide anion generating drugs such as DQ. Such an explanation does not hold for either HQ or BQ, which appear to be acting directly as free radical scavengers in these experiments. PMID- 8719802 TI - Exposure and characterization of the action of noradrenaline at dopamine receptors mediating endothelium-independent relaxation of rat isolated small mesenteric arteries. AB - 1. Previously, we reported that noradrenaline (NA), in addition to its alpha 1 adrenoceptor-mediated contractile effect, may relax the rat small mesenteric artery (SMA) in order to account for steep Schild plots obtained with compounds classified as alpha 1-adrenoceptor antagonists. In this study, a relaxant action of NA has been exposed in the rat isolated, endothelium-denuded SMA precontracted by the thromboxane A2-mimetic, U46619. 2. NA, but not the selective alpha 2 adrenoceptor agonist, UK14304, produced concentration-dependent contraction of the SMA (pEC50 = 5.7 +/- 0.1). After precontraction with 0.1 microM U46619, 10 nM 30 microM NA produced a further contraction (pEC50 = 6.1 +/- 0.2), while higher concentrations of NA produced small, but significant, relaxant responses. 3. In the presence of 1 microM prazosin, 0.1-30 microM NA produced concentration dependent relaxation (pIC50 = 5.9 +/- 0.1) after precontraction with 0.1 microM U46619. The NA relaxation concentration-effect curve was completely inhibited by 1 microM of the beta 1/beta 2-adrenoceptor antagonist, timolol. However, when the concentration of prazosin was increased by 10 fold (10 microM), NA once again produced concentration-dependent relaxation (pIC50 = 4.5 +/- 0.2). This relaxation concentration-effect curve was not blocked by a 10 fold higher concentration of timolol (10 microM), nor by the presence of idazoxan (10 microM), cyanopindolol (10 microM), NG-nitro-L-arginine methyl ester (L-NAME, 100 microM), indomethacin (10 microM) or sulpiride (1 microM). However, haloperidol (10 microM) and (+/-)-SCH-23390 (10 nM) produced significant inhibition of the relaxation, suggesting the involvement of dopamine D1 receptors. 4. Dopamine also produced concentration-dependent relaxation following U46619 precontraction (pIC50 = 5.4 +/- 0.1) which was significantly inhibited by haloperidol and (+) SCH-23390. Pretreatment with 10 microM phenoxybenzamine for 60 min produced a significant inhibition of the dopamine and NA relaxation curves and application of the operational model of agonism yielded estimates of the affinity (pKA = 5.3 +/- 0.2 and 4.4 +/- 0.2) and efficacy (log gamma = 0.06 +/- 0.11 and 0.01 +/- 0.10) for dopamine and NA, respectively, at D1 receptors. 5. HV723 (0.1 and 1 microM), a ligand that yielded a Schild plot slope parameter of unity as an antagonist of NA in the contractile assay, produced concentration-dependent inhibition of the NA-mediated relaxation (pA2 approximately 8). 6. The results of this study indicate that NA can activate D1 receptors mediating relaxation in the rat SMA at concentrations which were encountered in our previous receptor classification experiments using competitive alpha 1-adrenoceptor antagonists. PMID- 8719803 TI - Induction of L-arginine transport and nitric oxide synthase in vascular smooth muscle cells: synergistic actions of pro-inflammatory cytokines and bacterial lipopolysaccharide. AB - 1. The interactions between pro-inflammatory cytokines and bacterial lipopolysaccharide (LPS) on L-arginine transporter and inducible nitric oxide synthase (iNOS) activities were examined in rat cultured aortic smooth muscle cells. 2. LPS induced a concentration (0.01-100 micrograms ml-1) and time (8-24 h)-dependent stimulation of nitrite production which was accompanied by a parallel increase in L-arginine transport. 3. Unlike LPS, activation of smooth muscle cells with either interferon-gamma (IFN-gamma, 100 u ml-1), tumour necrosis factor-alpha (TNF-alpha, 300 u ml-1) or interleukin-1 alpha (IL-1 alpha, 100 u ml-1) failed to stimulate L-arginine transport or increase nitrite accumulation. 4. When applied in combination with LPS (100 micrograms ml-1) both IFN-gamma and TNF-alpha, but not IL-1 alpha, enhanced the effects observed with LPS alone. Furthermore, activation of cells with LPS and IFN-gamma had no effect on uptake of the neutral amino acid L-citrulline but selectively increased the Vmax for L-arginine transport 2.8 fold and nitrite levels from 24 +/- 7 to 188 +/ 14 pmol micrograms-1 protein 24 h-1. 5. The substrate specificity, Na- and pH independence of saturable L-arginine transport in both unactivated (K(m) = 44 microM, Vmax = 3 pmol micrograms-1 protein min-1) and activated (K(m) = 75 microM, Vmax = 8.3 pmol micrograms-1 protein min-1) smooth muscle cells were characteristic of the cationic amino acid transport system y+. 6. Cycloheximide (1 microM) abolished induction of L-arginine transport and nitrite accumulation in response to LPS and IFN-gamma. In contrast, the glucocorticoid dexamethasone (10 microM, 24 h) selectively inhibited nitrite production. 7. Our results demonstrate that pro-inflammatory mediators selectively enhance transport of L arginine under conditions of sustained NO synthesis by vascular smooth muscle cells. In addition, the differential inhibition of iNOS and L-arginine transporter activity by dexamethasone suggests that distinct signalling pathways mediate induction of the cationic transport protein and iNOS. The close coupling between substrate supply and NO production may have important implications in the pathogenesis of several disease states including endotoxin shock. PMID- 8719804 TI - Impaired endothelium-dependent relaxation in isolated resistance arteries of spontaneously diabetic rats. AB - 1. Previous studies have shown that endothelium-dependent relaxation in the aorta of spontaneously diabetic bio bred rats (BB) is impaired. 2. We have investigated noradrenaline (NA) contractility, endothelium-dependent acetylcholine (ACh) and bradykinin (BK) relaxation, and endothelium-independent sodium nitroprusside (SNP) relaxation in mesenteric resistance arteries of recent onset BB rats and established insulin treated BB rats, compared to their age-matched non diabetic controls. 3. There was no significant difference in the maximum contractile response or sensitivity to noradrenaline in either of the diabetic groups compared to their age-matched controls. 4. Incubation with the nitric oxide synthetase inhibitor NG-nitro-L-arginine (L-NOARG) resulted in a significant increase in maximum contractile response to noradrenaline in the recent onset age matched control group (P < 0.05). Analysis of the whole dose-response curve (using ANOVA for repeated measures with paired t test) showed a significant left ward shift following the addition of L-NOARG (P < 0.001). A similar but less marked shift (P < 0.01) was evident in vessels from recent onset diabetics. An overall shift in both sensitivity and maximum response was also evident in the age-matched non diabetic controls of the insulin-treated group (P < 0.05). However, by contrast, there was no significant change in sensitivity in the insulin-treated diabetic rats. 5. ACh-induced endothelium-dependent relaxation was significantly impaired in the recent onset diabetic rats compared to their age-matched controls (47 +/- 11% versus 92 +/- 2%, P < 0.05, n = 6), and in the insulin treated diabetic rats (34 +/- 5% versus 75 +/- 6%, P < 0.05, n = 6). The relaxation responses to BK also were significantly impaired in the diabetic rats compared to their age-matched controls (recent onset: 20 +/- 3% versus 72 +/- 7%, P < 0.05, n = 6; insulin treated: 12 +/- 9% versus 68 +/- 7%, P < 0.05, n = 7). 6. Incubation with either the nitric oxide synthetase substrate, U-arginine, or the free radical scavenging enzyme superoxide dismutase (150 mu ml-1) failed to improve the attenuated response of acetylcholine-induced relaxation in the diabetic vessels. 7. Endothelium-dependent relaxation mediated by ACh and BK was significantly attenuated in both the diabetic and control vessels after incubation with L-NOARG. 8. Pretreatment with a cyclo-oxygenase inhibitor, indomethacin, significantly enhanced the relaxation to ACh in both the recent onset and insulin treated diabetic rats (42 +/- 10%, n = 7 versus 64 +/- 7%, n = 7, P < 0.05, and 40 +/- 5%, n = 7 versus 65 +/- 9%, n = 6, P < 0.05). 9. Following endothelium removal, there was a marked impairment in endothelium dependent relaxation responses to ACh and BK in both the diabetic and control vessels. 10. Incubation with the thromboxane A2 receptor antagonist SQ29548, did not significantly improve the ACh endothelium-dependent relaxation response in the diabetic vessels. 11. Endothelium-independent relaxation to sodium nitroprusside was significantly impaired in the first group of diabetic vessels studied; however, subsequent studies showed no impairment of the sodium nitroprusside response in the diabetic vessels. 12. In conclusion, the ability of the endothelium to regulate vascular contractility is reduced in recent onset diabetic vessels, and significantly impaired in established insulin treated diabetics. Relaxation to the endothelium-dependent vasodilators ACh and BK was impaired in both the recent onset and the established insulin treated diabetics, and the ACh response was significantly improved following pretreatment with indomethacin, suggesting a role for a cyclo-oxygenase-derived vasoconstrictor. Preliminary studies with a thromboxane A2, receptor antagonist, SQ29548 did not significantly improve the impaired relaxation to ACh, indicating that the vasoconstrictor prostanoid is not thromboxane A2. PMID- 8719805 TI - Differential effects of potassium channel blockers on extracellular concentrations of dopamine and 5-HT in the striatum of conscious rats. AB - 1. The selective Ca(2+)-activated K+ channel blocker apamin increased extracellular 5-hydroxytryptamine (5-HT) concentrations in the striatum when administered through the microdialysis probe at doses of 0.1 mM and 1 mM. Extracellular dopamine concentrations increased only at the highest dose administered (1 mM). 2. Mast cell degranulating peptide (MCDP), which blocks the dendrotoxin sensitive delayed rectifier (DR) current, increased extracellular concentrations of dopamine at dose of 10 microM-100 microM but had no effect on 5 HT. 3. The non selective K+ channel blocker tetraethylammonium (TEA) induced a dose-dependent (1 mM-10 mM) increase in extracellular dopamine concentrations and an increase in 5-HT which showed little or no dose-dependency. 4. 4-Aminopyridine (4-AP), a blocker with some similar characteristics to MCDP, increased extracellular dopamine concentrations at doses of 10 microM-1 mM, but had no effect on 5-HT. 5. These findings suggest that dopamine release may be modulated by DR-like current and/or A-current K+ channels. However, in view of the similar effects of MCDP and 4-AP at the concentrations used it is more likely that the dendrotoxin-sensitive DR-like current is involved. In contrast, 5-HT release appears to be modulated by Ca(2+)-activated K+ channels. PMID- 8719806 TI - Postjunctional alpha 1- and beta-adrenoceptor effects of noradrenaline on electrical slow waves and phasic contractions of cat colon circular muscle. AB - 1. The postjunctional excitatory and inhibitory effects of noradrenaline and selective alpha 1- and beta-adrenoceptor agonists on electrical and mechanical activity of cat colon muscle strips were studied by microelectrode recordings and isometric force measurements. Experiments were performed in the presence of tetrodotoxin (0.5 microM) or atropine (0.5 microM). 2. Circular muscle cells near the submucosal border had a mean resting membrane potential of -76.1 +/- 1.2 mV and exhibited electrical slow waves at frequencies of 4-6 cycles min-1. The mean values of electrical slow wave components were: upstroke potential, -40.7 +/- 1.2 mV; plateau potential, -43.7 +/- 0.8 mV; and duration, 4.9 +/- 0.4 s. Electrical slow waves were in phase with rhythmic contractions of the circular muscle layer. Muscle cells near the myenteric border had a mean testing membrane potential of 51.1 +/- 5.5 mV and did not exhibit electrical slow waves. 3. Noradrenaline (1 microM) increased the duration of electrical slow waves. This effect was inhibited by prazosin (1 microM) and potentiated by propranolol (5 microM), indicating activation of alpha 1- and beta-adrenoceptors. Also, when alpha 1 adrenoceptors were irreversibly blocked by phenoxybenzamine (1 microM), noradrenaline decreased the duration of electrical slow waves. Phenylephrine (1 microM), a selective alpha 1-adrenoceptor agonist, and isoprenaline (1 microM), a beta-adrenoceptor agonist, increased or decreased the duration of electrical slow waves, respectively. 4. Phenylephrine (0.01-5 microM) caused a linear increase in the area of electrical slow waves and phasic contractions but did not affect resting membrane potential or resting muscle tension. Higher concentrations of phenylephrine (5-50 microM) depolarized the resting membrane potential (2-6 mV) and increased muscle tone. 5. Nitrendipine or verapamil (each at 5 microM) reduced the amplitude of the upstroke potential and nearly abolished the plateau phase of the electrical slow waves. In the presence of L-type Ca2+ antagonists, noradrenaline (1-10 microM) or phenylephrine (1-100 microM) had no effect on electrical slow waves and phasic contractions. This indicates that the effects of noradrenaline and phenylephrine involve the influx of extracellular Ca2+ through voltage-dependent L-type Ca2+ channels. 6. Ryanodine, an alkaloid that depletes intracellular Ca2+ stores nearly abolished phasic contractions. In muscle strips, pretreated with ryanodine (10 microM for 30 min), phenylephrine (1 microM) increased and isoprenaline (1 microM) decreased the duration of electrical slow waves but neither was able to reverse the ryanodine-suppressed phasic contractions. This suggests that adrenoceptor effects on electrical slow waves are coupled to contractions via Ca2+ release from ryanodine-sensitive intracellular stores. 7. In summary, noradrenaline activates postjunctional alpha 1- and beta-adrenoceptors. Activation of alpha 1-adrenoceptors increases the magnitude of electrical slow waves and phasic contractions, whereas activation of beta-adrenoceptors decreases them. The alpha 1-adrenoceptor mediated effects on electrical slow waves and phasic contractions require the influx of Ca2+ through voltage-gated L-type Ca2+ channels. Phasic contractions also involve Ca2+ release from ryanodine-sensitive intracellular stores. PMID- 8719807 TI - Anti-ataxic effects of TRH and its analogue, TA-0910, in Rolling mouse Nagoya by metabolic normalization of the ventral tegmental area. AB - 1. The mechanism of the anti-ataxic action of thyrotropin-releasing hormone (TRH) and its analogue. TA-0910, in the Rolling mouse Nagoya (RMN), an ataxic mutant mouse, has been investigated. 2. TRH (30 mg kg-1, i.p.) and TA-0910 (3 mg kg-1, i.p.) reduced the fall index (number of falls/spontaneous motor activity), an index of ataxia, 10-30 and 10-60 min after administration, respectively. 3. Relative local cerebral glucose utilization (LCGU) in the cerebellum and ventral tegmental area (VTA) of the rolling mouse was significantly smaller than that in normal animals. TRH (30 mg kg-1, i.p.) and TA-0910 (3 mg kg-1, i.p.) increased the relative LCGU value of the VTA but not of the cerebellum in rolling mice to the level of normal animals. 4. These results suggest that the ataxia of the rolling mouse may be due to dysfunction of the cerebellum and VTA, and that amelioration by TRH and TA-0910 could result from metabolic normalization of the VTA. PMID- 8719808 TI - [3H]-L-2-amino-4-phosphonobutyrate labels a metabotropic glutamate receptor, mGluR4a. AB - 1. The ligand binding site of subtype mGluR4a of the metabotropic glutamate receptor family was characterized by using [3H]-L-2-amino-4-phosphonobutyrate ([3H]-L-AP4) binding. 2. Specific [3H]-L-AP4 binding to membranes prepared from baby hamster kidney (BHK) cells transfected with a vector encoding mGluR4a accounted for 60-70% of the total binding whereas no specific binding of [3H]-L AP4 was observed to membranes prepared from BHK cells expressing the vector only. 3. Specific binding of [3H]-L-AP4 to mGluR4a was detectable at 0 degree C, was saturated with 10 min and enhanced by Cl(-)-ions but not by divalent cations (Mg2+, Ca2+, Mn2+). 4. [3H]-L-AP4 binding showed a maximal binding density (Bmax) of 3.0 +/- 0.5 pmol mg-1 protein and an affinity (KD) of 441 nM. A modest decrease in affinity was observed in the presence of 0.1 mM guanosine-5'-O-(3 thio)trisphosphate-gamma-S, the KD being 761 nM and the Bmax 3.4 +/- 0.6 pmol mg 1 protein. 5. The following rank order of affinity for mGluR4a was observed: L AP4 = L-serine-O-phosphate > glutamate = (2S,1S,2S)-2-(carboxycyclopropyl) glycine > 1-amino-3-(phosphonomethylene)cyclobitanecar-boxylate > > (1S,3R)-1 aminocyclopentane-1,3-dicarboxylate = quisqualate > ibotenate. 6. A highly significant correlation was observed between the potencies of the compounds to inhibit forskolin-stimulated cyclic AMP-formation in BHK cells expressing mGluR4a and the affinity for displacement of [3H]-L-AP4 binding from mGluR4a suggesting that this binding site is functionally relevant. 7. In conclusion, [3H]-L-AP4 is a suitable radioligand for characterizing mGluR4a when expressed in BHK cells. Interestingly, a significant correlation was found between the ability of various compounds to displace [3H]-L-AP4 binding from mGluR4a and the previously observed potencies for inhibition of synaptic transmission via L-AP4 sensitive glutamatergic pathways. These data support the hypothesis that the L-AP4 receptor is contained within the mGluR family. PMID- 8719809 TI - Attenuation by creatine of myocardial metabolic stress in Brattleboro rats caused by chronic inhibition of nitric oxide synthase. AB - 1. The present experiment was undertaken to investigate: (a) the effect of nitric oxide synthase (NOS) inhibition, mediated by oral supplementation of the NOS inhibitor, NG-nitro-L-arginine methyl ester (L-NAME), on measures of myocardial energy metabolism and function: (b) the effect of oral creatine supplementation on these variables, in the absence and presence of L-NAME. 2. In one series of experiments, 4 weeks oral administration of L-NAME (0.05 mg ml-1 day-1 in the drinking water) to Brattleboro rats caused significant reductions in myocardial ATP, creatine, and total creatine concentrations and an accumulation of tissue lactate when compared with control animals. Administration of creatine (0.63 mg ml-1 day-1 in the drinking water) for 4 weeks elevated myocardial creatine and total creatine concentrations and reduced lactate accumulation, but did not significantly affect ATP or phosphocreatine (PCr). Concurrent treatment with creatine and L-NAME prevented the reduction in creatine and total creatine concentrations, and significantly attenuated the accumulation of lactate and the reduction in ATP seen with L-NAME alone. 3. In a second series of experiments, 4 weeks treatment with L-NAME and creatine plus L-NAME increased mean arterial blood pressure in conscious Brattleboro rats. Hearts isolated from these animals showed decreased coronary flow and left ventricular developed pressure (LVDP), and total mechanical performance. Treatment with creatine alone had no measurable effect on either mean arterial blood pressure or coronary flow in isolated hearts. However, there was an increase in LVDP, but not in total mechanical performance, because there was a bradycardia. 4. These results indicate that creatine supplementation can attenuate the metabolic stress associated with L NAME administration and that this effect occurs as a consequence of the action of creatine on myocardial energy metabolism. PMID- 8719810 TI - Photosensitization of oesophageal smooth muscle by 3-NO2-1, 4-dihydropyridines: evidence for two cyclic GMP-dependent effector pathways. AB - 1. Photoactivated mechanical responses that resulted from exposure to 3-NO2-1,4 dihydropyridines (3-NO2-DHP5) or NO-donors were examined in rat isolated oesophageal smooth muscle with a view to determining the role of calcium and cyclic GMP. 2. Isometric contractile force was recorded in preparations bathed in normal Tyrode or 110 mM K(+)-depolarizing solution. Exposure to (+)-PN 202791, (+/-)-Bay K 8644 and (-)-PN 2020791 or the photodegradable NO-donors, sodium nitroprusside (SNP), streptozotocin (STZ) and sodium nitrite photosensitized precontracted tunica muscularis mucosae preparations in a concentration-dependent fashion. Photosensitizing potency followed the order: (+/-)-PN 202791 > (+/-)-Bay K 8644 > (-)-PN 202791 > SNP > STZ > NaNO2. 3. A low amplitude, slow photorelaxation (slope: 1 mg s-1) was obtained with the L-channel antagonists (-) PN 202791 and (+)-Bay K 4407. Photosensitization by the agonist enantiomers (+) PN 202 791 and (-)-Bay K 5407, as well as racemic Bay K 8644, was mimicked by NO donors and showed at least three different components, consisting of (i) a fast relaxation (slope: 140 mg s-1), (ii) a fast "off-contraction', and (iii) a delayed slow relaxation. The fast components, but not the delayed slow relaxation, were abolished by blockade of L-type voltage-operated calcium channels, chelation of extracellular calcium and skinning of the plasmalemma, suggesting their mediation by a process linked to calcium entry through L channels. 4. Both cyclopiazonic acid (3-30 microM) and ryanodine (30 microM) inhibited the fast response. This inhibition was accelerated in the presence of extracellular calcium and resembled that seen in tissues exposed to the calcium ionophore A 23187 (1 microM). In calcium depleted tissues, cyclopiazonic acid (3 microM) prevented restoration of the cis-dioxolane-induced contraction following re-exposure to a calcium containing high K+ buffer, but failed to inhibit the photoresponse. 5. Both the fast and slow relaxations were potentiated by zaprinast (10 microM) and inhibited by LY B3583 (10 microM). However, in calcium depleted, calyculin A-precontracted preparations only the slow relaxation was evident. 6. The present results support the conclusion that: (i) functional L channels are required for the expression of the fast components of the 3-NO2-DHP- or NO-donor-induced photoresponse, (ii) NO photorelease followed by activation of soluble guanylyl cyclase is responsible for the photosensitizing activity of 3 NO2-DHPs and (iii) regulation of the contractile proteins via cyclic GMP dependent phosphorylation may underlie the slow relaxation. PMID- 8719811 TI - The inhibitory effect of 3-amino-1,2,4-triazole on relaxation induced by hydroxylamine and sodium azide but not hydrogen peroxide or glyceryl trinitrate in rat aorta. AB - 1. In this study we investigated the role of catalase in relaxation induced by hydroxylamine, sodium azide, glyceryl trinitrate and hydrogen peroxide in isolated rings of rat aorta. 2. Hydrogen peroxide (1 microM-1 mM)-induced concentration-dependent relaxation of phenylephrine (PE)-induced tone in endothelium-containing rings. In endothelium-denuded rings, however, higher concentrations (30 microM-1 mM) of hydrogen peroxide were required to produce relaxation. The endothelium-dependent component of hydrogen peroxide-induced relaxation was abolished following pretreatment with N(O)-nitro-L-arginine methyl ester (L-NAME, 30 microM). L-NAME (30 microM) had no effect, however, on hydrogen peroxide-induced relaxation in endothelium-denuded rings. 3. Pretreatment of endothelium-denuded rings with catalase (1000 u ml-1) blocked relaxation induced by hydrogen peroxide (10 microM-1 mM). The ability of catalase to inhibit hydrogen peroxide-induced relaxation was partially blocked following incubation with 3-amino-1,2, 4-triazole (AT, 50 mM) for 30 min and completely blocked at 90 min. 4. Pretreatment of endothelium-denuded rings with methylene blue (MeB, 30 microM) inhibited relaxation induced by hydrogen peroxide (10 microM-1 mM), sodium azide (1-300 nM), hydroxylamine (1-300 nM) and glyceryl trinitrate (1-100 nM) suggesting that each acted by stimulation of soluble guanylate cyclase. 5. Pretreatment of endothelium-denuded rings with AT (1-50 mM, 90 min) to inhibit endogenous catalase blocked relaxation induced by sodium azide (1-300 nM) and hydroxylamine (1-300 nM) but had no effect on relaxation induced by hydrogen peroxide (10 microM-1 mM) or glyceryl trinitrate (1-100 nM). 6. In a cell-free system, incubation of sodium azide (10 microM-3 mM) and hydroxylamine (10 microM 30 mM) but not glyceryl trinitrate (10 microM-1 mM) with catalase (1000 u ml-1) in the presence of hydrogen peroxide (1 mM) led to production of nitrite, a major breakdown product of nitric oxide. AT (1-100 mM) inhibited, in a concentration dependent manner, the formation of nitrite from azide in the presence of hydrogen peroxide. 7. These data suggest that metabolism by catalase plays an important role in the relaxation induced by hydroxylamine and sodium azide in isolated rings of rat aorta. Relaxation appears to be due to formation of nitric oxide and activation of soluble guanylate cyclase. In contrast, metabolism by catalase does not appear to be involved in the relaxant actions of hydrogen peroxide or glyceryl trinitrate. PMID- 8719812 TI - The contribution of tumour necrosis factor-alpha and endothelin-1 to the increase of coronary resistance in hearts from rats treated with endotoxin. AB - 1. Inflammatory disease states predispose to myocardial infarction. Here we have investigated the effects of a systemic inflammatory response syndrome, i.e. lipopolysaccharide (LPS)-induced circulatory shock in rats, on coronary vascular tone. 2. Anaesthetized rats were given LPS (10 mg kg-1, i.v.) and the hearts excised 180 min later for isolated perfusion at constant flow by the Langendorff technique. Once the ex vivo perfusion was started, the perfusion pressure strongly increased in these hearts compared to hearts from control rats (130 +/- 3 vs. 49 +/- 3 mmHg after 10 min). This increase in coronary resistance was not associated with a reduction in endothelial cell function, for the vasodilator responses to bradykinin were unchanged. 3. When hearts were removed 30 min after the injection of LPS, the LPS-induced rise in perfusion pressure was delayed. No changes in perfusion pressure were seen when the hearts were removed 15 min after the injection of LPS. Pre-treatment with cycloheximide or an anti-tumour necrosis factor-alpha (TNF-alpha) antibody or continuous infusion in vivo and in vitro of the endothelin ETA receptor selective antagonist FR 139317, greatly decreased the increase in coronary vascular resistance induced by LPS. 4. These data suggest that TNF-alpha may induce the release of endothelin-1 (ET-1) and that this mediates at least part of the coronary vasoconstriction. This hypothesis is supported by the demonstration that LPS administration increased the circulating levels of both TNF-alpha and ET-1. 5. We conclude, therefore, that in inflammatory disease states, such as LPS-induced septic shock, there is the sequential release of TNF-alpha and endothelin-1 which leads to an increase in coronary vascular tone and so a predisposition to myocardial ischaemia. Inactivation of TNF-alpha by an antibody as well as ETA receptor blockade by a selective antagonist may effectively interfere with this pathway. PMID- 8719813 TI - The effect of endotoxin on sympathetic responses in the rat isolated perfused mesenteric bed; involvement of nitric oxide and cyclo-oxygenase products. AB - 1. The effects of endotoxin on the vasoconstrictor responses to sympathetic nerve stimulation (SNS) were investigated in the rat isolated perfused mesenteric bed. 2. Rats received either saline (0.1 ml h-1) or endotoxin (2.5 mg kg-1 h-1) intravenously for 4 h; the mesenteric beds were then isolated, perfused with Krebs and prepared for SNS (50 V, 3 ms, 7-40 Hz). 3. SNS caused a frequency dependent vasoconstrictor response which was abolished by either tetrodotoxin (10(-7) M), prazosin (2.4 x 10(-7) M) or guanethidine (2.4 x 10(-7) M). 4. In mesenteric vascular beds removed from rats infused with endotoxin, there were markedly impaired vasoconstrictor responses to SNS, although responses to noradrenaline were not modified. 5. Removal of the endothelium with distilled water prevented endotoxin-induced impairment of vasoconstrictor responses to SNS, without modifying these responses in preparations from control rats. 6. Pretreatment with dexamethasone (3 mg kg-1 i.p. 1h before commencing endotoxin or saline infusions) did not modify responses to SNS in control rats but prevented the effects of endotoxin. 7. Both L-NAME (10(-3) M) and indomethacin (10(-5) M) restored responses to SNS in preparations from endotoxin-treated rats without modifying these responses in control preparations. However, co-administration of L-NAME and indomethacin markedly augmented responses in both control and endotoxin-treated preparations. 8. The effects of L-NAME were reversed by addition of L-arginine (10(-3) M). 9. The data suggest that endotoxin impairs the release of noradrenaline and that this effect is secondary to increased production of nitric oxide and prostanoids, possibly by the endothelium. PMID- 8719814 TI - Structure-activity relationships for a series of phenylglycine derivatives acting at metabotropic glutamate receptors (mGluRs). AB - 1. The actions of a series of twelve phenylglycine derivatives at metabotropic glutamate receptors (mGluRs) linked to both phosphoinositide hydrolysis (PI) and cyclic AMP were investigated. 2. PI hydrolysis was determined by the accumulation of [3H]-inositol-monophosphate ([3H]-IP1) in neonatal ral cortical slices prelabelled with [3H]-myo-inositol. The non-selective mGluR agonist (1S,3R)-1 aminocyclopentane-1, 3-dicarboxylic acid ((1S,3R)-ACPD) produced a concentration dependent increase in [3H]-IP1 (EC50 approximately 20 microM). This agonist was subsequently used to investigate potential antagonist activity of the phenylglycine derivatives. Of the compounds tested (+)-alpha-methyl-4 carboxyphenylglycine (M4CPG) and (RS)-alpha-ethyl-4-carboxyphenylglycine (E4CPG) were the most active with KP values of 0.184 +/- 0.04 mM and 0.367 +/- 0.2 mM respectively. 3. Activity at adenylyl cylase-coupled mGluRs was investigated by determining the accumulation of [3H]-cyclic AMP in adult rat cortical slices. [3H]-cyclic AMP accumulation, elicited by 30 microM forskolin, was inhibited by (2S,3S,4S)-alpha-(carboxycyclopropyl)glycine (L-CCG-1) and L-2-amino-4 phosphonobutanoate (L-AP4) with respective EC50 values of 0.3 microM and 10 microM. Neither agonist was able to inhibit completely forskolin stimulated cyclic AMP accumulation; this is evidence that not all adenylyl cyclase is susceptible to modulation by mGluRs. Phenylglycine derivatives were examined for their ability to antagonize the inhibition of [3H]-cyclic AMP accumulation by L CCG-1 or L-AP4 at their EC50 concentrations. 4. A rank order of potency of the phenylglycine derivatives as antagonists of L-AP4 and L-CCG-1 was obtained. The most effective compound. (RS)-alpha-methyl-3-carboxymethylphenylglycine (M3CMPG) had IC50 values in the order of 1 microM against L-AP4 and 0.4 microM against L CCG-1. 5. The results from this study indicate that phenylglycine-derived compounds can discriminate between groups of metabotropic glutamate receptors and may also display some selective activity between subtypes within groups. Future work based on these findings may lead to the development of more selective and potent compounds as important pharmacological tools. PMID- 8719815 TI - Characterization of prejunctional 5-HT receptors mediating inhibition of sympathetic vasopressor responses in the pithed rat. AB - 1. It has recently been shown that continuous infusions of 5-hydroxytryptamine (5 HT) are able to inhibit, in a dose-dependent manner, the pressor responses induced by preganglionic (T7-T9) sympathetic stimulation in pithed rats pretreated with desipramine (50 micrograms kg-1, i.v.). This inhibitory effect, besides being significantly more pronounced at lower frequencies of stimulation (0.03-I Hz) and devoid of tachyphylaxis, is reversible after interrupting the infusions of 5-HT (up to 5.6 micrograms kg-1 min-1). In the present study we have characterized the pharmacological profile of the receptors mediating the above inhibitory effect of 5-HT. 2. The inhibition induced by 5.6 micrograms kg-1 min-1 of 5-HT on sympathetically-induced pressor responses was not blocked after i.v. treatment with physiological saline (1 ml kg-1), ritanserin (0.1 mg kg-1), MDL 72222 (0.15 mg kg-1) or tropisetron (3 mg kg-1), which did not modify the sympathetically-induced pressor responses per se, but was significantly antagonized by the 5-HT1-like and 5-HT2 receptor antagonist, methysergide (0.3 mg kg-1), which also produced a slight attenuation of the pressor responses to 0.03 and 0.1 Hz per se. 3. Unexpectedly and contrasting with methysergide, the 5-HT1 like and 5-HT2 receptor antagonists, methiothepin (0.01, 0.03 and 0.1 mg kg-1) and metergoline (1 and 3 mg kg-1), apparently failed to block the above 5-HT induced inhibition. Nevertheless, it is noteworthy that these antagonists also blocked the electrically-induced pressor responses per se, presumably by blockade of vascular alpha 1-adrenoceptors and, indeed, this property might have masked their potential antagonism at the inhibitory 5-HT1-like receptors. 4. Consistent with the above findings, 5-carboxamidotryptamine (5-CT, a potent 5-HT1-like receptor agonist), metergoline and methysergide mimicked the inhibitory action of 5-HT with the following rank order of agonist potency: 5CT > > 5-HT > metergoline > or = methysergide. 5. Taken together, the above results suggest that the inhibitory action of 5-HT on the electrically-induced pressor responses is primarily mediated by an action on inhibitory prejunctional 5-HT1-like receptors leading to a decrease in the sympathetic nerve discharge. Interestingly, 5-HT induced excitatory mechanisms could be made manifest once the inhibitory action of 5-HT had been antagonized. PMID- 8719817 TI - Evidence for a projection from the perireticular thalamic nucleus to the dorsal thalamus in the adult rat and ferret. AB - During early development, the perireticular thalamic nucleus is very large (i.e. has many cells) and has a strong projection to the dorsal thalamus and to the cerebral neocortex. By adulthood, the nucleus has much reduced in size and only a few cells remain. It is not clear whether these perireticular cells that remain into adulthood maintain their connections with the dorsal thalamus and with the neocortex. This study examines this issue by injecting neuronal tracers into various nuclei of the dorsal thalamus (dorsal lateral geniculate nucleus, medial geniculate complex, ventroposteromedial nucleus, lateral posterior nucleus, posterior thalamic nucleus) and into different areas of the neocortex (somatosensory, visual, auditory). After injections of tracer into the individual nuclei of the rat and ferret dorsal thalamus, retrogradely-labelled perireticular cells are seen. In general, after each injection, the retrogradely-labelled perireticular cells lie immediately adjacent to a group of retrogradely-labelled reticular cells. For instance, after injections into the medial geniculate complex, perireticular cells adjacent to the auditory reticular sector are retrogradely-labelled, whilst after an injection into the dorsal lateral geniculate nucleus, retrogradely-labelled perireticular cells adjacent to the visual reticular sector are seen. By contrast, injections of tracer into various areas of the rat and ferret neocortex result in no retrogradely-labelled cells in the perireticular nucleus. Thus, unlike during perinatal development when perireticular cells project to both neocortex and dorsal thalamus, perireticular cells in the adult seem to project to the dorsal thalamus only: the perireticular projection to the neocortex appears to be entirely transient. PMID- 8719816 TI - Different effects of fibrates on the microsomal fatty acid chain elongation and the acyl composition of phospholipids in guinea-pigs. AB - 1. The effects in vitro and in vivo of three fibric acid derivatives, clofibrate (CFB), bezafibrate (BFB) and gemfibrozil (GFB) on some enzyme activities related to fatty acid biosynthesis, namely palmitoyl-CoA synthetase and hydrolases (microsomal and cytosolic), NADH and NADPH cytochrome c reductases and acyl-CoA elongases were investigated in guinea-pigs. 2. The three fibrates inhibited acyl CoA elongation in vitro, irrespective of the substrate of elongation used (saturated, monounsaturated, polyunsaturated) and with an order of potency GFB > BFB > CFB. In the case of GFB, inhibition occurred at concentrations that can be reached in vivo. 3. Microsomal palmitoyl-CoA hydrolase and synthetase were also inhibited in vitro (GFB > or = BFB > CFB), whereas NADH cytochrome c reductase activity was increased by GFB. Nevertheless, the magnitude of changes were lower than those observed in elongation activities. 4. Treatment with fibrates did not produce peroxisomal proliferation in guinea-pigs, as measured by peroxisomal beta oxidation activity and liver weight/body weight ratio. Nevertheless, fibrates provoked a reduction in plasma cholesterol and triglycerides, at least in GFB- and BFB-treated animals. 5. Fatty acid elongation was significantly modified by GFB treatment in vivo. The remaining enzyme activities studied were only slightly changed by fibrate treatment. 6. Treatment with BFB and to a lesser extent with CFB, increased the relative proportion of MUFA (palmitoleic and oleic acids) in microsomal phospholipids, whereas PUFA (mainly linoleic acid) decreased. GFB behaved differently, increasing palmitic and linoleic acids and decreasing stearic and oleic acids. The latter changes are attributable to an inhibition of elongation activity by GFB. 7. The changes observed after fibrate treatment in both rats and guinea-pigs, as they are not directly related to peroxisome proliferation, could be more reliably extrapolated to man than those observed only in rats. PMID- 8719818 TI - S-laminin and N-acetylgalactosamine located at the synaptic basal lamina of skeletal muscle are involved in synaptic recognition by growing neurites. AB - The purpose of the work reported here is to identify molecular components of the synaptic basal lamina of skeletal muscle fibres which allow recognition of original synaptic sites by regenerating motor axons. We focused on s-laminin and components recognized by the lectin Dolichos biflorus agglutinin previously shown to be specifically located at the synaptic basal lamina. We used a cryoculture bioassay in which chick ciliary ganglion neurons grow on rat skeletal muscle cryostat sections. In control cultures, neurites extended over the muscle sections in close association with the muscle cell surface. It was observed that most of the neurites that extended towards the endplate zone and reached an area of 40 microns around the neuromuscular junction ceased to grow when they contacted the synaptic site. Masking either lectin receptors or some s-laminin molecule epitopes prior to the culture of neurons alters the behaviour of growing neurites. On sections treated either with Dolichos biflorus agglutinin or anti s laminin monoclonal antibodies (D5 and C4) most of the neurites did not stop their growth at the synaptic regions. Moreover, treating muscle sections with Dolichos biflorus agglutinin removed the gradient of substratum affinity around the endplate. These results indicate that the s-laminin and Dolichos biflorus agglutinin receptors present on muscle cell surfaces may play a functional role in the interaction of growing neurites with original synaptic sites in the process of neuromuscular regeneration. PMID- 8719819 TI - Immunocytochemical localization of the GTP-binding protein G0 alpha in the vestibular epithelium and ganglion of the guinea-pig. AB - The guanine nucleotide binding protein G0 alpha was immunolocalized in the guinea pig vestibular system by confocal and electron microscopy. The vestibular sensory epithelia consist of the macula utriculi, macula sacculi and cristae ampullaris of the semicircular canals. Two types of hair cells are present in these epithelia. Type I hair cells are surrounded by an afferent nerve calyx that receives efferent innervation and type II hair cells are innervated directly by the afferent and efferent nerves. G0 alpha protein was observed on the inner face of the afferent calyceal membrane surrounding type I hair cells and in nerve endings in contact with type II hair cells. No labelling was found in the stereocilia and cuticular plate of type I and type II hair cells whereas the cytoplasmic matrix displayed a diffuse labelling. The plasma membrane of the supporting cells showed discreet labelling in the confocal microscope that are still confirmed by electron microscopy. A positive reaction was also observed along the plasma membrane of the vestibular ganglion neurons. Immunoblotting with affinity-purified polyclonal rabbit antibodies selective for the 39 kDa alpha subunit of G0 indicated that G0 alpha protein was present in both the vestibular ganglion. That G0 alpha labelling was observed in the cytoplasm of vestibular hair cells and in nerve endings contacting hair cells suggests that G0 may be involved in the modulation of vestibular neurotransmission. PMID- 8719821 TI - Cytoplasmic constriction and vesiculation after axotomy in the squid giant axon. AB - The squid giant axon responded to a transection injury by producing a gradient of cytoplasmic and vesicular changes at the cut end. At the immediate opening of the cut axon the cytoplasm was fragmented and dispersed and the vesicles in this region were in rapid Brownian movement. Approximately 0.1 mm further in, at the site of maximal axonal constriction, the axoplasm was condensed into a compact, constricted mass containing many large vesicles. The axoplasm was normal a few millimetres beyond this constricted, vesiculated end. It appears that transection triggered the transformation of normal axoplasm into a tightly constricted, highly vesiculated structure. This modified axoplasm at the cut end may slow the spread of damage and degeneration by preventing the bulk outflow of axoplasm, by slowing down the loss of intracellular molecules and by slowing down the influx of destructive extracellular ions (like calcium and chloride). PMID- 8719820 TI - Cytochemical evidence for redistribution of membrane pump calcium-ATPase and ecto Ca-ATPase activity, and calcium influx in myelinated nerve fibres of the optic nerve after stretch injury. AB - There has been controversy for some time as to whether a posttraumatic influx of calcium ions occurs in stretch/nondisruptively injured axons within the central nervous system in both human diffuse axonal injury and a variety of models of such injury. We have used the oxalate/pyroantimonate technique to provide cytochemical evidence in support of such an ionic influx after focal axonal injury to normoxic guinea pig optic nerve axons, a model for human diffuse axonal injury. We present evidence for morphological changes within 15 min of injury where aggregates of pyroantimonate precipitate occur in nodal blebs at nodes of Ranvier, in focal swellings within axonal mitochondria, and at localized sites of separation of myelin lamellae. In parallel with these studies, we have used cytochemical techniques for localization of membrane pump Ca(2+)-ATPase and ecto Ca-ATPase activity. There is loss of labelling for membrane pump Ca(2+)-ATPase activity on the nodal axolemma, together with loss of ecto-Ca-ATPase from the external aspect of the myelin sheath at sites of focal separation of myelin lamellae. Disruption of myelin lamellae and loss of ecto-Ca-ATPase activity becomes widespread between 1 and 4 h after injury. This is correlated with both infolding and retraction of the axolemma from the internal aspect of the myelin sheath to form periaxonal spaces which are characterized by aggregates of pyroantimonate precipitate, and the development of myelin intrusions into invaginations of the axolemma such that the regular profile of the axon is lost. There is novel labelling of membrane pump Ca(2+)-ATPase on the cytoplasmic aspect of the internodal axolemma between 1 and 4 h after injury. There is loss of an organized axonal cytoskeleton in a proportion of nerve fibres by 4-6 h after injury. We suggest that these changes demonstrate a progressive pathology linked to calcium ion influx after stretch (non-disruptive) axonal injury to optic nerve myelinated fibres. We posit that calcium influx, linked to or correlated with changes in Ca(2+)-ATPase activities, results in dissolution of the axonal cytoskeleton and axotomy between 4 and 6 h after the initial insult to axons. PMID- 8719822 TI - Reinnervation of rat Pacinian corpuscles after nerve crush during the postcritical period of development. AB - The ultrastructure of crural Pacinian corpuscles was examined after sciatic nerve crush performed in 7- to 20-day-old rats, i.e. during the postcritical period of development when the corpuscles no longer degenerate after axotomy but cease growing. The aim of our study was to assess the innervation pattern and structural changes of the corpuscles following transient denervation and subsequent reinnervation during their maturation and growth. Reinnervated corpuscles were examined by electron microscopy from 2.5 months after nerve crush onwards. After sciatic nerve crush at 7 days of age, the corpuscles are mostly reinnervated with multiple axon terminals, each of them enclosed within a newly formed inner core. The axial multiple cores are in part covered by a layer of concentric inner core lamellae and surrounded by a capsule, both structures having survived from the original corpuscle. After nerve crush at 10 days of age, reinnervated Pacinian corpuscles usually contain, in their axial region, a denervated remainder of the original core together with a few regenerated axon terminals enclosed within new inner cores. These axial structures are surrounded by a layer of concentric lamellae of the original core which may accommodate some regenerated terminals. Additional axon terminals with their small inner cores may be found at the outer aspect of the composite core beneath the capsule. When the nerve is crushed in 15-day-old rats, the inner core which is already well developed remains preserved by the time of reinnervation, and regenerating axons grow in between the original lamellae inducing only moderate neoformation of 2-3 lamellar layers which enclose the terminals. After crushing the sciatic nerve in 20-day-old rats, formation of new inner core lamellae is minimal and regenerated terminals become accommodated between the original lamellar of the core as is the case in adult animals. Regeneration of new inner cores and reinnervation of the preserved lamellar structure thus characterize the recovery of Pacinian corpuscles following reinnervation after nerve crush during the postcritical period of their development. PMID- 8719823 TI - Axon-glial relationships in the anterior medullary velum of the adult rat. AB - The anterior medullary velum is a thin sheet of CNS tissue which roofs the rostral part of the IVth ventricle and contains fascicles of myelinated fibres which, in part, arise from the nucleus of the IVth cranial nerve. This study used histochemical, immunohistochemical, and intracellular dye-injection techniques to describe cellular interrelationships in the velum in whole-mounts and in sections. Rip antibody-stained whole mounts provided a unique description of both oligodendrocyte units (defined as an oligodendrocyte and the complement of myelinated internodal segments it forms), and consecutive myelin sheaths along the same axon. A broad range of unit morphologies was categorised into four arbitrary groups, according to classical criteria, which comprised small cells supporting the short, thin myelin sheaths of 15-30 small diameter axons (Type I), through intermediate types (II & III), to the largest cells forming the long, thick myelin sheaths of 1-3 large diameter axons. Rip antibody and ferric ion ferrocyanide staining, together with intracellular dye injection, revealed oligodendrocyte process branching patterns and their mode of engagement of myelin sheaths, nodes of Ranvier, and the spatial disposition of the outer cytoplasmic rims of myelin sheaths. The latter formed a conspicuous spiral ridge on the exterior surface of myelin sheaths which connected with the paranodal loops at each heminode. Large bundles of axons decussated through the velum, the bulk of which were IVth nerve fibres which constituted the IVth nerve rootlet. The PNS/CNS transitional zone of the IVth nerve was located 0.25-0.50 mm along the root, where astrocytic end-feet defined an abrupt margin, convex towards the periphery, where the heminodes of central and peripheral myelin were apposed, and where the basal lamina tubes of the Schwann cell units were discontinued. The basal processes of ependymal cells lining the ventricular wall of the velum, passed between axon bundles before abutting on the basal lamina of the pia. Many of these processes branched and ran along the axonal bundles. A monolayer of microglia occupied a subependymal stratum in which the non-overlapping dendritic territories of each cell formed a regular mosaic throughout the velum without any obvious interaction with either axons or other glial cells. Astrocytes were also uniformly distributed; their fine processes made up a dense lattice amongst axons, often running parallel and within the fibre bundles; stouter ones had terminal end-feet which undercoated the basal lamina of both the glia limitans externa and the blood vessels in the velum. PMID- 8719824 TI - Transcription of a new zinc finger gene, rKr1, is localized to subtypes of neurons in the adult rat CNS. AB - Proteins which share zinc finger DNA binding motifs comprise one of the main families of transcription factors. We have previously described rKr1, a new rat Cys2/Hys2 zinc finger gene of the Kruppel gene family. This gene is predominantly expressed in the nervous system, with highest abundance in neurons and with lower abundance in developing oligodendrocytes of the CNS. Here, we have undertaken a detailed anatomical analysis of rKr1 expression in the adult brain of the rat using in situ hybridization. Our results show that rKr1 is expressed in a specific manner in defined subpopulations of neurons in many regions of the adult brain. Moderate levels of rKr1 mRNA were detectable in some structures of the telencephalon (e.g. cerebral cortex and hippocampus) and a few nuclei of the thalamus. The highest degree of labelling was seen in both upper and lower motor neurons of the mesencephalon and rhombencephalon (e.g. red nucleus, gigantocellular reticular nuclei, motor nuclei of the cranial nerves). High levels of rKr1 expression were also present in spinal motoneurons and dorsal root ganglion cells. In order to determine if rKr1 gene expression can be regulated, we have examined the expression pattern of rKr1 in the facial nucleus in response to facial nerve lesion. The expression of rKr1 in the facial nucleus showed a differential downregulation, reaching lowest levels 1 week after transection of the facial nerve. By 3 weeks after lesion, expression of rKr1 on the operated side of the brain reached normal levels and was identical to that of the unoperated side. These data suggest that rKr1 could be involved in the maintenance of the phenotypic differentiation of specific neuronal subtypes including motoneurons. PMID- 8719825 TI - Gene therapy for brain tumours. PMID- 8719826 TI - Arteriovenous malformations of the posterior fossa: a report on 28 cases and review of the literature. AB - Posterior fossa arteriovenous malformations are uncommon lesions accounting for between 7 and 18% of all intracranial arteriovenous malformations. They have a greater incidence of haemorrhage and a higher morbidity and mortality if untreated compared with those localized in the supratentorial compartment. We report our experience with 28 cases of posterior fossa arteriovenous malformations referred to the senior author between January 1971 and December 1993. The anatomy, symptomatology, treatment and results are discussed with regard to the most recent literature. PMID- 8719827 TI - Revisional lumbar microdiscectomy: an analysis of operative findings and clinical outcome. AB - Forty-one consecutive patients who underwent a revision microlumbar exploration for recurrent or persistent sciatica were reviewed retrospectively to analyse the operative findings and assess the clinical outcome following surgery. Thirty three (80%) patients were found to have a recurrent intervertebral disc protrusion at the previous site, two patients had a disc herniation at a new site, one had severe perineural scarring, two had lateral recess stenosis, one patient had undergone previous exploration at an incorrect site and in two patients no cause for ongoing symptoms was found. Nineteen of the 33 patients with a re-prolapse presented with persistent or recurrent sciatica within 1 year of their first operation. The other 14 patients presented with a late re-prolapse (after 1 year) and their clinical outcome was better than for those patients with an early re-prolapse (12/14 vs 11/19 satisfactory result, respectively). The result of operating on patients with a late re-prolapse was comparable to the 80 95% satisfactory outcomes following primary lumbar microdiscectomy reported by other authors. PMID- 8719828 TI - Nasal complications of the direct transnasal approach to the pituitary fossa. AB - The transnasal trans-sphenoidal approach to the pituitary fossa is direct and fast. This study is aimed at establishing the nasal morbidity arising from this approach. One of the 25 patients developed secondary haemorrhage and another suffered recurrent minor epistaxis. The peak nasal inspiratory flow rates showed no significant difference between the left and right anterior nasal passages. Adhesions were noted in seven patients and no septal perforations were found. Three patients, who had no previous nasal symptoms, developed postoperative unilateral nasal obstruction, two secondary to left septal deviation and the other caused by right-sided anterior nasal space adhesions. The study suggests that the operation has a low nasal morbidity. PMID- 8719830 TI - Haemodynamic consequences of embolizing aneurysms: a transcranial Doppler study. AB - Twenty patients with aneurysms were studied with transcranial Doppler before, during and after endovascular treatment with Guglielmi detachable coils. Catheterization of the anterior circulation decreased middle cerebral artery velocities by 15-20%. Inserting coils into the aneurysms increased pulsatility indices modestly, reflecting a stiffening of the cerebral circulation. Pre- and posttreatment velocities were not significantly different, and there was no evidence of coil embolization precipitating vasospasm. The haemodynamic stability, seen during and after therapy, suggests that coil embolization may prove a safe alternative way of treating acutely ruptured aneurysms. PMID- 8719829 TI - Middle fossa decompression of the trigeminal sensory root for trigeminal neuralgia--a re-appraisal and a modification of the technique. AB - Twenty-three patients with intractable trigeminal neuralgia were treated by a modified technique of middle fossa extradural decompression of the trigeminal sensory root at the petrous ridge. After exposing the ganglion and the root at the petrous ridge, by an extradural Frazier's approach a thin layer of autogenous fat was interposed between the dura propria and the middle fossa dura anteriorly, and between the petrous ridge and the root posteriorly. The fat layers were used to prevent dural adhesions and to maintain a space between the petrous ridge and the root. The dura propria was preserved intact. All patients obtained immediate relief of pain following surgery. Twenty patients continue to remain pain free. Two patients have mild occasional pains and one has a persistent pain of moderate intensity. One patient developed a dense sensory loss and two had delayed facial weakness which recovered completely within a month. The mechanism of trigeminal neuralgia is discussed on the basis of anatomical, histopathological and electrophysiological findings and the rationale of middle fossa decompression of the root is stressed. This method is short, simple and safe and may serve as an alternative to the more recent methods of treating trigeminal neuralgia. PMID- 8719831 TI - Surgery for intractable epilepsy secondary to viral encephalitis. AB - In a series of 668 craniotomy cases for intractable epilepsy, in 11 patients (1.6%) the presumed aetiology was a previous episode of viral encephalitis, a widespread cerebral inflammation, which may therefore produce multifocal epilepsy. Seven patients had chronic ictal electrocorticography (ECoG). Two of these had a generalized onset, one bilateral temporal independent onset, and four unilateral mesial temporal onset. Three patients underwent corpus callosotomy. Eight had resections (seven anterior temporal lobectomy (ATL), and one ATL and frontal resection). Pathology was hippocampal sclerosis in four, neocortical gliosis in three and one specimen was normal. Of the resection patients, three (37%) were seizure-free at a mean of 3 years postoperatively and three (37%) unchanged. It is concluded that where the aetiology of intractable epilepsy is viral encephalitis the possibility of multifocal epilepsy should be considered. Chronic ECoG is recommended. Nevertheless, unilateral hippocampal sclerosis can also be produced and the results of ATL in these cases can be rewarding. PMID- 8719832 TI - Results of the lateral rhinotomy approach in transsphenoidal microsurgery for growth hormone-secreting pituitary adenoma. AB - The results of transsphenoidal microsurgery for growth hormone (GH)-secreting pituitary adenomas in a series of 47 patients are presented. A modified transsphenoidal approach with lateral rhinotomy was employed for the access to the sphenoid sinus. Remission, defined as mean GH concentration less than 2.5 ng/ml, was achieved in 79% of the patients. Macroadenoma size of the tumour and a GH concentration more than 50 ng/ml were factors unfavourable for the outcome. The experience of the lateral rhinotomy approach in transsphenoidal surgery of acromegaly has been favourable. The distance from the incision to the sella is shorter than in rhinoseptal approaches, where the depth of the operative field is determined by the distance from the nasal spine to the sella. The lateral rhinotomy also allowed an exposure of the sella of adequate width in all patients. These aspects of the procedure facilitated adenoma removal. PMID- 8719833 TI - The significance of traumatic intraventricular haemorrhage in severe head injury. AB - Since the advent of computed tomography (CT) traumatic intraventricular haemorrhage (IVH) has been diagnosed more often. It has reportedly been associated with a poor prognosis, but pure or solitary IVH is rare, suggesting that other lesions occurring concurrently with it may contribute to the poor outcome. In a series of 65 patients with severe head injury (GCS < or = 8), 14 (22%) had IVH on initial CT. Death rate in these 14 was 21%, not significantly different from that in patients without IVH (14%), although a significantly higher proportion of patients without IVH had a good outcome. These results suggest that mortality is related to other lesions associated with IVH rather than to IVH alone and that the presence of IVH does not necessarily lead to a poor outcome. PMID- 8719834 TI - Hemispherectomy for intractable seizures: a further modification and early experience. AB - Hemispherectomy is well recognized as an effective treatment for some types of intractable epilepsy, but the procedure is also known to be associated with significant late complications. A number of modifications to the original operation have been developed to try and minimize such late complications. These are reviewed and a further modification is described using a Zenoderm graft to isolate the hemispherectomy cavity from the remaining CSF pathways. We present our early experience with this new modification in seven patients with intractable epilepsy. PMID- 8719835 TI - Tentorial dural flap for transtentorial surgery. AB - For a subtemporal transtentorial approach to the anterior part of the posterior fossa a tentorial dural flap is described. An incision is begun at the free edge of the tentorium at the level of the posterior edge of the cerebral peduncle and is then directed anterolaterally towards the superior petrosal sinus. A triangular flap of the tentorial dura is everted over the superior petrosal sinus. The flap has the advantage of being simple and safe, and provides a wide window to the infratentorial structures. In addition the occasional troublesome bleeding from the superior petrosal sinus and the petrosal vein is avoided. The fourth and fifth cranial nerves are protected and exposed widely. PMID- 8719836 TI - Spinal cord compression caused by ossification of the transverse ligament of the atlas. AB - Although ossification of the posterior longitudinal ligament and ligamentum flavum are well known, ossification of the transverse ligament of the atlas is extremely rare. We present the case of a 79-year-old man who developed a gradually progressive spastic quadriparesis caused by upper cervical canal stenosis due to ossification of the transverse ligament of the atlas together with ligamentum flavum hypertrophy. PMID- 8719837 TI - Perils of a 'true' posterior communicating artery aneurysm. AB - Successful 'clipping' of the apparent neck of a posterior communicating artery aneurysm was carried out, but 2 days later the patient had a further haemorrhage and died. Postmortem examination revealed that the aneurysm was that of a rare fusiform 'true' posterior communicating artery aneurysm. PMID- 8719838 TI - Intracranial angiolipoma: report of two cases. AB - Intracranial tumours composed of adipose tissue and abnormal blood vessels are rarely recognized. We report two such cases; one showed the radiological and histological appearances of a combined lipoma and an arteriovenous malformation, the other the appearances of a lipoma with a prominent capillary component. Computed tomography did not give any indication of their vascular nature, but surgery was complicated by intra- and postoperative haemorrhage. PMID- 8719839 TI - Symptomatic Rathke's cleft cyst with hypophysitis. AB - We report a case of symptomatic Rathke's cleft cyst (RCC) with associated chronic hypophysitis. Symptomatic RCCs are rare and an associated inflammatory reaction is even rarer. The clinical and pathological findings are discussed in the light of previous published reports, together with the problems involved in making a preoperative diagnosis in such cases. PMID- 8719840 TI - Thrombosis of the straight sinus complicating hormone replacement therapy. AB - Oral contraceptives have been implicated in the pathogenesis of dural venous thrombosis. No similar association has been confirmed between postmenopausal hormone replacement therapy and cerebral venous thrombosis, although one previous report in the literature has identified such a case. We present a further case of thrombosis of the straight sinus in a patient on hormone replacement therapy (HRT) and suggest that dural sinus thrombosis is a rare, but dangerous complication of HRT. PMID- 8719841 TI - Glossopharyngeal neuralgia associated with vascular compression and choroid plexus papilloma. AB - We report two cases of presumed idiopathic glossopharyngeal neuralgia that were discovered intraoperatively to be associated with compression by choroid plexus papillomas, and by a variable degree of vascular compression at the root entry zones of cranial nerves IX and X. The combination of the two entities in glossopharyngeal neuralgia has not previously been reported. PMID- 8719842 TI - Unusual CT appearances with a supratentorial meningioma. AB - The majority of intracranial meningiomas are isodense, well-marginated masses, adjacent to a dural surface. They show intense uniform enhancement following intravenous contrast. Atypical appearances can occur. Hypodense meningiomas have been described due to necrosis, cystic change or lipidisation of the tumour. A meningioma is described which showed two unusual features on computed tomography a uniformly hypodense mass on an unenhanced scan and only minimal contrast enhancement. Extensive lipidisation of the tumour was demonstrated histologically. PMID- 8719843 TI - Investigation of some biochemical and functional effects of cryopreservation of human spermatozoa using an automated freezing-quick-thawing method. AB - The objective of the present studies was to assess the functional integrity of the sperm plasma membrane and metabolic and motility characteristics of the recovered motile fraction of human spermatozoa subjected to an automated freezing/quick-thawing method. Sperm membrane features examined included progesterone-induced changes in intracellular levels of calcium ([Ca2+]i), as measured by the fluorescent fura-2 indicator, and the tight binding of spermatozoa to homologous zonae pellucidae as assessed by the hemizona assay (HZA). Basal [Ca2+]i intracellular adenosine triphosphate (ATP) and adenosine diphosphate (ADP) levels determined using chemiluminescence with luciferin luciferase, and motility parameters determined using a computer-aided semen analyser (CASA) were studied concomitantly as an expression of metabolic/functional status. Ejaculates from fertile men (donors) were evaluated after swim-up separation of the motile fraction in both fresh and cryopreserved thawed samples, and fractions of each ejaculate (fresh and frozen-thawed) were subjected to parallel measurements of the same parameters at the same time frame. Basal and progesterone-induced increase in [Ca2+]i, and ATP levels (up to 24 h) were similar in fresh and frozen-thawed samples. HZA results showed a modest (26%) although significant (p = 0.008) decrease in binding in frozen-thawed samples. The ratios of ATP/ADP in fresh and frozen-thawed samples were also found to be similar. Although post-thaw sperm motility was significantly lower than that of the fresh samples, comparison of the results indicated that the method was capable of preserving > 65% of motile spermatozoa in almost all of the samples cryopreserved. Additionally, the swim-up rescued a motile fraction in the frozen-thawed samples that was not significantly impaired with regard to motility, mean linear velocity or linearity as compared to the fresh fractions in the first 4 h. Our results show that this automated freezing-quick-thawing method results in a small reduction in sperm-zona binding capacity, and that the time dependent decline in motility parameters observed for both fresh and cryopreserved-thawed samples cannot be related to ATP deficiency under the conditions of our experiments. These in-vitro results are coincident with the maintenance of fertilizing capacity for donor spermatozoa in the in-vitro fertilization (IVF) setting. PMID- 8719844 TI - Stimulation of movement and acrosome reaction of human spermatozoa by PC12 liposomes encapsulating ATP. AB - The effect of co-incubating human spermatozoa with 8 mmol/L dilauroylphosphatidylcholine (PC12) liposomes containing 6 mmol/L adenosine 5' triphosphate (LATP) was assessed by CASA and compared to that obtained with blank PC12 liposomes (LB). The aim of this study was to investigate if such treatments can improve sperm movement and sustain sperm motility over time. Significant and similar increases in straight-line velocity and linearity of sperm movement in B2 capacitating medium (both p < 0.01) were obtained with LB and LATP treatments (final concentration: 0.38 mmol/L PC12 and 0.5 mmol/L ATP) while in Tyrode's medium supplemented with 10 mg/mL BSA, these movement parameters were increased significantly only in sperm aliquots treated with LATP. Furthermore, after incubation for 0.5 h in Tyrode's, a bioluminescence assay of intracellular ATP indicated no significant change in ATP concentration for LATP-treated spermatozoa while the ATP content of control and LB-treated spermatozoa decreased significantly during the same period (both p < 0.05). The effect of liposomes on the acrosome reaction was also investigated jointly with CASA. These experiments were performed by fluorescence microscopy, using PSA-FITC and the supravital stain Hoechst 33258. After a precapacitation period of 3 h in BWW medium the spermatozoa were incubated for 1 h with LATP, LB, LB+free ATP and free ATP alone (final concentration 0.5 mmol/L ATP). Under these conditions the percentage of acrosome-reacted spermatozoa was increased similarly after LATP and LB treatments compared to control (respectively from 4.9 to 12%, p < 0.01 and 4.9 to 11.3%, p < 0.05) but the percentage of true acrosome-reacted spermatozoa, and the values for all movement characteristics (except percentage motility) were increased significantly only with LATP treatment. The results indicate the potential of PC12 vesicles for introducing highly hydrophilic compounds into spermatozoa, as well as for modulating membrane structures and functions required for fertilization. PMID- 8719845 TI - Immunohistochemical quantitative study of the peritubular lamina propria after induction of testicular atrophy induced by epinephrine. AB - Changes in the testicular peritubular lamina propria in rats treated for 1-11 weeks with intra-scrotal injections of epinephrine were studied by quantitative immunohistochemical methods. In control testes, BrdU-labelled nuclei (proliferating cells) were observed only in spermatogonia and some primary spermatocytes, whereas testes from epinephrine-treated rats showed BrdU labelling in some of the spermatogonia and in peritubular cells. Immunostaining for transforming growth factor beta 1 (TGF-beta 1) was present in germ cells, Sertoli cells and Leydig cells; vimentin immunostaining was found mainly in Sertoli cells; desmin immunostaining was found in the peritubular cells, and immunostaining for type IV collagen, laminin and fibronectin was found in the extracellular matrix of the lamina propria. The volume densities of seminiferous tubules (including seminiferous epithelium, lamina propria and tubular lumen) that immunostained for TGF-beta 1, vimentin, laminin, desmin or fibronectin were calculated. All of these parameters increased significantly in testes from epinephrine-treated animals during the course of the experiment, except for desmin immunostaining which showed no significant change in volume density. Since total seminiferous tubule volume decreased markedly in the testes of treated rats during the experiment, the transformation of relative values for immunostaining into absolute volumes per testis revealed a significant increase in TGF-beta 1 immunostaining, no significant change in vimentin immunostaining, and a significant decrease in desmin immunostaining during the time of the study. The absolute volume occupied by laminin and fibronectin immunostaining decreased from the 3rd to the 8th weeks of treatment, and increased from the 8th to the 11th weeks. These changes, associated with germ cell depletion and tubular fibrosis, suggest that tubular ischaemic atrophy caused by epinephrine alters the peritubular myoid cells, which change immunophenotype and increase their secretion of the extracellular matrix components producing tubular fibrosis. The mechanism of this alteration may involve direct effects on the peritubular cells or the changes may be secondary to germ cell and/or Sertoli cell lesions. PMID- 8719846 TI - Effect of glyceryl trinitrate on sperm motility and lipid peroxidation in normozoospermic men. AB - The objective of this study was to assess the in-vitro effects of the nitric oxide substrate glyceryl trinitrate (GTN) on sperm motility and membrane lipid peroxidation. Nitric oxide (NO) can impair sperm motility, possibly by an alteration of cyclic nucleotide levels. NO may also be protective against lipid peroxidation. Semen samples from nine normospermic men were prepared by a swim-up technique. Each specimen was divided into four aliquots, one of which was the control sample. The other three had 10(-6), 10(-8) or 10(-10) M GTN added. Sperm motility was then analysed over 180 min using a Hamilton Thorn motility analyser. Lipid peroxidation was assessed by measuring media malondialdehyde (MDA) levels at 180 min. Compared with control, the following measurements were reduced (p < 0.05) over the first 60 min in the 10(-6) M GTN aliquots only: mean path velocity (reduced by 14-15%), curvilinear velocity (reduced by 12-21%), straight-line velocity (reduced by 18-19%) and percentage of hyperactivated spermatozoa (reduced by 38-43%). MDA levels and head movement parameters were comparable amongst all aliquots (p > 0.05). The depressant effects of GTN on sperm motility appeared to be transient and reversible. The effects observed may be due to NO generated by GTN, or to GTN itself. This suggests that NO may have a role in vivo as a physiological inhibitor of sperm motility. The addition of GTN did not appear either to cause sperm membrane damage or to protect the spermatozoa from lipid peroxidation. PMID- 8719847 TI - Incidence of testicular mononuclear cell infiltrates in normal human males and in patients with germ cell neoplasia. AB - The incidence of mononuclear cell (MNC) infiltrates was studied in the testes of the following: 45 patients with carcinoma in situ (CIS) as the only lesion in the testis, CIS accompanied by an early invasion of CIS or CIS accompanied by invasive germ cell tumour (GCT) of the testis; 100 men and 146 boys who had died suddenly and unexpectedly, and 100 infertility patients. The results suggest that (1) the incidence of MNC infiltration increases with increasing severity of testicular malignant changes, (2) increased MNC infiltration is also evident in the contralateral testis where no malignant cells can be observed, (3) the incidence of MNC infiltration in men who died suddenly is not different from that in men who have had their testes biopsied because of infertility, and (4) the incidence of MNC infiltration is high in testes of boys younger than 1 year of age. PMID- 8719848 TI - Hydrogen hexachloroplatinate induces the acrosome reaction in human spermatozoa. AB - Human spermatozoa from healthy donors (n = 7) were washed in Tyrode's medium containing 0.6 mg/ml freeze-dried egg yolk, filtered through glass wool and exposed to 0.5, 5, 50, 500 or 1000 microM hydrogen hexachloroplatinate. Viability, membrane integrity and the acrosome reaction were examined using trypan blue exclusion, hypo-osmotic swelling test and fluoresceinated Pisum sativum agglutinin, respectively, after incubation for 3 or 6 h at 37 degrees C. While sperm motility, viability and membrane integrity were not affected by the platinum compound after incubation for 3 h, the number of acrosome-reacted spermatozoa increased from 16.0 +/- 6.4% (control, mean +/- SEM) to 21.0 +/- 3.3 (0.5 microM), 22.3 +/- 4.3% (5 microM), 28.0 +/- 4.3% (50 microM, p < 0.01), 29.3 +/- 3.9% (500 microM, p < 0.01) and 43.9 +/- 7.4% (1 mM, p < 0.001); a further increase was detected after incubation for 6 h. However, the percentages of dead and immotile spermatozoa and those with defective membranes were also higher, suggesting that the acrosome reaction was caused by degenerative processes after long-term incubation. In conclusion, hydrogen hexachloroplatinate does not affect sperm motility, membrane integrity or viability, but it does induce the acrosome reaction after incubation for 3 h before cytotoxic effects are measurable. Similar effects of halide salts of platinum on receptor-mediated exocytosis have been described in other cells such as mast cells and basophils in vivo and in vitro. PMID- 8719850 TI - Statement on the general reduction in sperm quality. AB - Over a period of 17 years, 360 young healthy men presented themselves as candidate donors for the sperm bank of the University Hospital of Ghent. Semen characteristics of these men were assessed using unaltered methods, and analysis was performed on the first sample delivered before any donor selection was performed. Sperm motility and morphology, but not the total sperm count per ejaculate, exhibited a highly significant decrease. Over 40% of candidate donors since 1990 exhibited subnormal sperm characteristics, as against only 5% of the group investigated before 1980. PMID- 8719849 TI - Male germ cell transplantation in rats: apparent synchronization of spermatogenesis between host and donor seminiferous epithelia. AB - Primordial germ cells (PGC) and gonocytes from male Sprague-Dawley rat fetuses and neonates were transplanted via the rete testis into the lumen of the seminiferous tubules of recipient adult Long Evans rats. The donor germ cells apparently differentiated into mini-tubules or irregular segments of seminiferous epithelium within the lumen of the host seminiferous tubules, and exhibited qualitatively normal spermatogenesis in 10 out of 16 recipients. The stage of spermatogenesis of the intraluminal epithelium was synchronized closely with that of the adjacent seminiferous tubule epithelium, suggesting that the spermatogenic cycle is regulated locally by the intraluminal microenvironment. Male germ cell transplantation provides an interesting new tool for investigating the control of spermatogenesis. PMID- 8719851 TI - Genetic aspects of human infertility. AB - Mutations of the human genome associated with the phenotype of infertility are well known. Some of these are visible under the microscope as specific chromosome mutations. Others arise from genes which function only in the germ line, or which function during development of the gonads, or which function also in non-gonadal somatic cells, in which cases sterility is manifest as a pleiotropic effect of dysfunction of this gene also in certain gonadal cells. Examples of mutations are presented in this paper. They indicate that men with a severe idiopathic sterility factor have an especially high risk for a genetically determined barrier to reproduction. PMID- 8719852 TI - Mitochondrial differentiation during meiosis of male germ cells. AB - In male germ cells mitochondria undergo dramatic morphological changes during spermatogenesis, at least three different types of mitochondrion being present. The usual cristae type of mitochondrion in spermatogonia, preleptotene and leptotene spermatocytes develops, via an intermediate form in zygotene spermatocytes, to the condensed form with almost no cristae which is typical of pachytene spermatocytes and early spermatids. In cell culture experiments in which isolated preparations of meiotic germ cells were used, it was shown that condensed mitochondria in pachytene spermatocytes cultured in Earle's minimal essential medium dedifferentiated to the intermediate type, while Sertoli cell conditioned medium (SC-CM) was able to maintain the condensed structure. SC-CM was also able to induce conversion of the intermediate type to the condensed type in isolated zygotene spermatocytes. Preliminary biochemical characterization showed the involvement of one or several proteinaceous factors > 10 kDa (PMMF: paracrine mitochondria maturation factor) that were protease (subtilisin)- and heat-sensitive. Three mitochondrial proteins served as markers for germ cells in different phases of maturation. The chaperonin hsp60 was detectable in the orthodox-type mitochondria of spermatogonia and primary spermatocytes (leptotene and zygotene). An ATP- dependent mitochondrial matrix enzyme -- the Lon-protease - appeared in the orthodox and intermediate forms of mitochondria in leptotene and zygotene spermatocytes. Sulphydryl oxidase is present in the condensed mitochondria of pachytene spermatocytes and early spermatids. PMID- 8719853 TI - NMR spectroscopy in andrology: research uses and possible clinical applications. AB - Research uses and possible clinical applications of NMR spectroscopy are considered. By monitoring extracts of testes using 31P-NMR spectroscopy, detailed analysis of high-energy phosphates, or pathways such as phospholipid biosynthesis or glycolysis, is possible. By using in-vivo 31P NMR spectroscopy of testes in animal experiments it has proved possible to discriminate non-invasively between experimentally impaired and control testes. It has been demonstrated in several human studies that discrimination between normal testes and those with impaired spermatogenesis and obstruction is possible by non-invasive NMR spectroscopy. In studies of semen, seminal plasma or spermatozoa, NMR spectroscopy is of high experimental value for analysis of sample composition, the kinetics of metabolites or the structure of single components, but clinical applications have not yet been clearly defined. PMID- 8719854 TI - Molecular mechanisms of the interaction between sperm and the zona pellucida in mammals: studies on the pig. AB - Research carried out on mammalian fertilization is summarized here. The main focus is on current knowledge of porcine gamete interaction. Attention has been given to the structural characterization of zona pellucida glycoproteins with sperm receptor activity and the primary zona pellucida-binding proteins of the spermadhesin family on spermatozoa, and to the role of acrosin as a secondary ligand for sulphated glycoproteins of the zona pellucida. Established similarities and differences between mammalian species are described to highlight the existence of both common and species-specific mechanisms. PMID- 8719855 TI - Clinical relevance of scrotal and transrectal ultrasonography in andrological patients. AB - Sonography of the scrotal contents, the prostate and the seminal vesicles has become an important diagnostic tool in andrology. In a prospective study of 1048 consecutive patients attending the Institute of Reproductive Medicine only 520 patients (49.6%) did not show sonographic abnormalities of the scrotal contents. The leading abnormality in the other 528 patients (50.4%) was a varicocele (194 patients, 18.5%), increased size of the epididymis (147 patients, 14.0%), epididymal cyst or spermatocele (55 patients, 5.2%), hydrocele (104 patients, 9.9%), testicular non-homogeneity (92 patients, 8.8%), testicular hypoechogenicity (132 patients, 12.6%), testicular cyst (12 patients, 1.1%) and testicular tumour (5 patients, 0.5%) (sum of percentages exceeds 50.4% because of multiple abnormalities in individual patients). In addition to the high incidence of pathological findings, the significantly higher incidence of testicular tumours compared to the general population and the early detection by ultrasonography render scrotal sonography a diagnostic procedure with high clinical relevance. Transrectal sonography of the prostate and seminal vesicles is valuable for detection of chronic urogenital infections or functional abnormalities of the seminal vesicles in infertile patients. In hypogonadal patients, transrectal examination of the prostate should be performed longitudinally to monitor the biological efficacy of testosterone treatment by measuring prostate growth and, in combination with palpation and PSA measurements, to screen for prostate cancer. PMID- 8719856 TI - The significance of computer-assisted semen analysis (CASA) for diagnosis in andrology and fertility prognosis. AB - The estimation of seminal parameters by CASA improves the objectivity, precision and reproducibility of values measured. Novel parameters, such as sperm velocity and characteristics of track direction, become measurable. An important advantage is direct on-line documentation. The biological significance of CASA parameters with respect to specific types of motility pattern or to fertility prognosis, however, is questionable. An important field is the analysis of pharmacological and toxicological influences on sperm motility. The evaluation of sperm morphology by CASA is not yet an established method. PMID- 8719857 TI - Advantages of strict (Tygerberg) criteria for evaluation of sperm morphology. AB - The purpose of this paper was to compare the validity of strict criteria for sperm morphology evaluation with other evaluation criteria in the diagnosis and prognosis of male fertility potential. Adoption of strict criteria is a holistic approach to sperm morphology evaluation which uses optimal preparation and evaluation procedures and criteria for a morphologically normal spermatozoan based on biological evidence. If liberal evaluation criteria are used, two sperm populations will be included in the normal population, namely those that are truly normal and an abnormal population. It was therefore postulated that results of strict criteria evaluation should provide better correlations with functional tests and fertilization outcome. In most studies where strict criteria was compared to WHO criteria it was found that strict criteria was a better prognosticator of expected in-vitro fertilization rates than WHO criteria. With regard to between- and within-observer correlation coefficients and coefficients of variation for repeated evaluations, results obtained using strict criteria compared favourably with or were better than results reported in the literature. Normal morphology, as evaluated by strict criteria, was also highly predictive of the outcome of certain functional tests, such as the hemizona assay and acrosin activity. From these and other data presented in this paper it can be concluded that sperm morphology evaluated according to strict criteria has definitive advantages over the other (liberal) criteria evaluation methods in the prediction of expected in-vivo and especially in-vitro fertilization rates. PMID- 8719858 TI - Immunological aspects of subfertility. AB - No significant relationship between semen quality [determined by standard sperm analysis and sperm-cervical mucus (CM) interaction testing in vivo and in vitro] and the presence of antisperm antibodies (ASA) [determined using three different methods] in serum samples of males and/or females of subfertile partnerships was found. These circulating ASA were not associated with reduced fertility in prospective studies. On the other hand, ASA in semen, particularly those of the IgA class, significantly impaired the ability of spermatozoa to penetrate CM and the fertility prognosis of the couple. ASA in CM must also be considered as a severe infertility factor but are a very rare cause of poor mucus quality. PMID- 8719859 TI - Acrosome reaction and fertility. AB - The acrosome reaction is important for fertilization. To diagnose this reaction the simple gelatin lysis test, the triple-stain technique or labelling with lectins can be used. The role of pentoxifylline in enhancing the acrosome reaction remains to be determined. PMID- 8719860 TI - Survey of medical therapy in andrology. AB - Despite considerable basic research effort, the therapeutic possibilities to improve impaired male fertility are still limited. This is partly because the cause of reduced fertility is unknown in many cases and, consequently, no specific therapy is available. Approaches to andrological therapy at different levels include stimulation of spermatogenesis, improvement of epididymal function or sperm transport within the male genital tract, and stimulation of sperm metabolism. Medical therapy of male fertility disturbances has to distinguish between specific and empirical treatment procedures. Specific treatment is based on a pathophysiological concept and implies accurate patient selection; the therapeutic success is predictable. In contrast, empirical treatment involves no patient selection according to specific criteria; thus, the therapeutic success is unpredictable. Hormonal treatment is the main choice in andrological therapy, including pulsatile administration of gonadotrophin-releasing hormone (GnRH), human gonadotrophins, anti-oestrogens and testosterone. Non-hormonal treatment is carried out using different pharmaceutical compounds such as alpha sympathomimetics, antibiotics, antiphlogistics, corticosteroids, mast cell blockers, pentoxifylline, kallikrein, captopril, vitamins and zinc. In patients with severe male factor who are resistant to therapy, procreation problems may be solved by the method of intracytoplasmic sperm injection (ICSI). PMID- 8719861 TI - Microsurgical aspects of the treatment of azoospermia. The BMFT Study Group for Microsurgery. AB - Technical advances in microsurgery have produced a calculable success rate for reconstructive surgery in cases of obstructive azoospermia. Nevertheless, in standardized vasovasostomy and tubulovasostomy, the different outcomes for patency and pregnancy indicate that further comprehensive clinical and basic scientific studies are required to improve the results of surgery. The aspiration of epididymal and testicular spermatozoa in combination with intra-cytoplasmic sperm injection (ICSI) is one example of a fruitful integration of microsurgery into new areas of reproductive medicine. Where there is ejaculatory duct obstruction, the continuity with the posterior urethra can be restored by a transurethral operation. PMID- 8719862 TI - Varicocele: indications for treatment. AB - Varicocele is detected in about one-third of men with subfertile semen quality. Treatment by means of transcatheter embolization using the tissue adhesive Histoacryl improves Sertoli cell function with increased concentrations of transferrin in semen, and a significant increase in sperm concentration, normal morphology and linear velocity. Failure to attain pregnancy after varicocele treatment occurs if pre-treatment sperm quality is normal, or in the case of azoospermia with elevated blood FSH levels or obstruction, or when there is severe coincidental pathology impairing fertility of the male or female partner. In all other cases, the poor results of varicocele treatment reported in some studies can only be explained by inadequate techniques. Varicocele detection and embolization treatment at puberty or during adolescence should be advocated to prevent deterioration of testicular function. PMID- 8719863 TI - Is there still a place for intra-uterine insemination as a treatment for male subfertility? A review. AB - Using the results of randomized and controlled studies, it was assessed whether there is still a place for intra-uterine insemination (IUI), with or without controlled ovarian hyperstimulation (COH), as a treatment for male subfertility. It is concluded that, in the case of a moderate to severe semen defect, well timed IUI should be applied in natural cycles only, because COH does not further improve treatment outcome. In contrast, in the case of a mild semen defect, the present evidence indicates that COH improves the probability of conception. If COH is applied, stimulation should be mild in order to prevent multiple pregnancies and the ovarian hyperstimulation syndrome. PMID- 8719864 TI - Statement on intra-uterine insemination. AB - Treatment of couple infertility due to male subfertility by means of intra uterine insemination (IUI) gives better results, in terms of per cycle and total cumulative pregnancy rate, if sperm preparation is performed using a discontinuous Percoll gradient than if centrifugation-resuspension is used. Also, the minimal semen requirements for successful IUI are lower with the former technique. Optimal epididymal function, with total alpha-glucosidase activity in seminal plasma > 83 IU/mL or Schorr stain result > 60%, is associated with a high probability of success of IUI [odds ratio (OR) = 11.1 and 9.4 respectively; p < 0.01]. If semen contains > 2.3 million white blood cells per mL or more than 13 million spermatozoa/mL with grade a motility the success rate is decreased (OR = 0.25 and 0.30 respectively; p < 0.05 and p < 0.01). It is concluded that IUI is a highly successful treatment in specific cases of male subfertility, provided that the correct technique of sperm preparation is used. PMID- 8719865 TI - New approaches to male infertility: IVF and microinjection. AB - The male factor represents a major problem in the treatment of infertility. New methods of assisted fertilization such as partial zona dissection (PZD), subzonal sperm injection (SUZI) and intracytoplasmic sperm injection (ICSI) are described and discussed in terms of their potential benefits for infertility treatment in comparison to classical in-vitro fertilization (IVF) and intra-uterine insemination (IUI). PMID- 8719866 TI - Minimal criteria of sperm quality for insemination and IVF therapy. AB - Minimal criteria are poorly established because prognosis of success for an individual couple is possible only statistically. In the past, therefore, there was wide variation in the minimal semen criteria required to ensure that therapy for male subfertility had a chance of being successful. The aim of this study was to define minimal criteria for semen parameters in husbands of women undergoing IVF and insemination at the Department of Obstetrics and Gynaecology, University of Gottingen. The results prove that the establishment of minimal semen criteria is only possible to a limited extent. At least 10 x 10(6) spermatozoa/ml, of which at least 30% are motile and 15% have progressive motility, are required for IVF or insemination therapy, despite the fact that pregnancies can be achieved with lower parameters. As a minimum, 20% of spermatozoa should be of normal morphology. On average, a curvilinear velocity of 35 microns/sec and a straight line velocity of 15 microns/sec are needed. It is questionable, however, if in future these minimal criteria will still be relevant. The successful injection of one spermatozoon into an oocyte (intracytoplasmic sperm injection, ICSI) suggests that all the semen quality parameters which have been demanded in the past are no longer of any value. Fertilization and pregnancy can be achieved by ICSI even with immotile spermatozoa, spermatozoa which have not undergone the acrosome reaction, spermatozoa without tails, and morphologically aberrant and/or immature spermatozoa. PMID- 8719867 TI - Indications for and results of intracytoplasmic sperm injection (ICSI). AB - In this report the indications for, and the results of, 1275 consecutive ICSI cycles carried out between October 1991 and December 1993 are described. Failure of fertilization in at least one previous IVF cycle, semen parameters below the threshold for standard IVF treatment and successful MESA or TESE procedure performed in patients with obstructive or non-obstructive azoospermia, respectively, were the indications in these ICSI cycles. In 1194 cycles, ejaculated spermatozoa were used, whereas 59 and 17 cycles were performed with epididymal and testicular spermatozoa, respectively. The normal fertilization rate was significantly higher with ejaculated spermatozoa than with epididymal or testicular spermatozoa, but no differences were observed with regard to embryo quality, the percentages of transfer after ICSI and the clinical pregnancy rates in the three groups of patients. PMID- 8719868 TI - Application of in-vitro fertilization in (almost) fertile couples should, and can, be avoided. AB - In-vitro fertilization is a successful treatment for infertile couples. However, in a minority of cases it can be dangerous, both for mother and child. IVF should therefore be avoided in (almost) fertile couples. A simple prognostic model makes it possible to distinguish couples with a good chance of conceiving spontaneously from those who deserve treatment in due course. Use of this model should enable IVF to be avoided in (almost) fertile couples. PMID- 8719869 TI - Statement on the ethical aspects of assisted fertilization. PMID- 8719870 TI - Regulation of yolk degradation, or how to make sleepy lysosomes. PMID- 8719871 TI - Matrix metalloproteinases are obligatory for the migration of preosteoclasts to the developing marrow cavity of primitive long bones. AB - A key event in bone resorption is the recruitment of osteoclasts to future resorption sites. We follow here the migration of preosteoclasts from the periosteum to the developing marrow cavity of fetal mouse metatarsals in culture, and investigate the role of proteinases and demineralization in this migration. Our approach consisted in testing inhibitors of proteinases and demineralization on the migration kinetics. Migration was monitored by histomorphometry and the (pre)osteoclasts were identified by their tartrate resistant acid phosphatase (TRAP) activity. At the time of explantation, TRAP+ cells (all mononucleated) are detected only in the periosteum, and the core of the diaphysis (future marrow cavity) consist of calcified cartilage. Upon culture, TRAP+ cells (differentiating progressively into multinucleated osteoclasts) migrate through a seam of osteoid and a very thin and discontinuous layer of mineral, invade the calcified cartilage and transform it into a "marrow' cavity; despite the passage of maturing osteoclasts, the osteoid develops into a bone collar. The migration of TRAP+ cells is completely prevented by matrix metalloproteinase (MMP) inhibitors, but not by a cysteine proteinase inhibitor, an inhibitor of carbonic anhydrase, or a bisphosphonate. The latter three drugs inhibit, however, the resorptive activity of mature osteoclasts at least as efficiently as do the MMP inhibitors, as assessed in cultures of calvariae and radii. Furthermore, in situ hybridizations reveal the expression of 2 MMPs, gelatinase B (MMP-9 or 92 kDa type IV collagenase) in (pre)osteoclasts, and interstitial collagenase (MMP-13) in hypertrophic chondrocytes. It is concluded that only MMPs appear obligatory for the migration of (pre)osteoclasts, and that this role is distinct from the one MMPs may play in the subosteoclastic resorption compartment. We propose that this new role of MMPs is a major component of the mechanism that determines where and when the osteoclasts will attack the bone. PMID- 8719872 TI - Localization of myosin II A and B isoforms in cultured neurons. AB - Tension generated by growth cones regulates both the rate and the direction of neurite growth. The most likely effectors of tension generation are actin and myosins. We are investigating the role of conventional myosin in growth cone advance. In this paper we report the localization of the two most prominent isoforms of brain myosin II in growth cones, neurites and cell bodies of rat superior cervical ganglion neurons. Affinity purified polyclonal antibodies were prepared against unique peptide sequences from human and rat A and B isoforms of myosin heavy chain. Although each of these antibodies brightly stained nonneuronal cells, antibodies to myosin heavy chain B stained neurons with greater intensity than antibodies to myosin heavy chain A. In growth cones, myosin heavy chain B was most concentrated in the margin bordering the thickened, organelle-rich central region and the thin, actin-rich peripheral region. The staining colocalized with actin bundles proximal and distal to the marginal zone, though the staining was more prominent proximally. The trailing edge of growth cones and the distal portion of the neurite often had a rimmed appearance, but more proximal regions of neurites had cytoplasmic labelling. Localizing MHC-B in growth cones previously monitored during advance (using differential interference contrast microscopy) revealed a positive correlation with edges at which retraction had just occurred and a negative correlation with lamellipodia that had recently undergone protrusion. Cell bodies were brightly labelled for myosin heavy chain B. Myosin heavy chain A staining was dimmer and its colocalization with filamentous actin bundles in growth cones was less striking than that of myosin heavy chain B. Growth cones stained for both myosin heavy chain A and B revealed that the two antigens overlapped frequently, but not exclusively, and that myosin heavy chain A lacked the elevation in the marginal zone that was characteristic of myosin heavy chain B. The pattern of staining we observed is consistent with a prominent role for myosin heavy chain B in either generating tension between widely separated areas of the growth cone, or bundling of actin filaments, which would enable other motors to effect this tension. These data support the notion that conventional myosin is important in growth cone advance and turning. PMID- 8719873 TI - Elucidation of nucleus-cytoplasm interaction: change in ability of the nucleus to express sexuality according to clonal age in Paramecium. AB - The expression of sexual activity in Paramecium caudatum is repressed for about 50 fissions after conjugation. The ability of the macronucleus to affect the expression of sexual activity according to the age of clonal development was investigated using the macronuclear fusion-reorganization method. When a mature macronucleus was transplanted into an immature cell and fused with a macronucleus in the immature cell, the clones derived from the recipient showed sexual immaturity. In a reverse experiment, an immature macronucleus was transplanted into a mature cell, and the clones also showed sexual immaturity. The ability of the macronucleus to transform mature cells to immature cells was clonal age dependent. The characteristics of the immature-mature hybrid macronucleus indicate that the immature macronucleus is dominant over the mature macronucleus with respect to the ability to express sexual activity. On the other hand, in cells of the early immaturity period, the micronucleus, known as the germ nucleus, shows the ability to undergo meiosis and eventually to produce progeny under control of the mature macronucleus. The expression of sexual activity is thought to be governed by the clonal age of the macronucleus and not by the clonal age of the micronucleus or cytoplasm. The macronucleus seems to determine the ability to express sexual activity by counting post-conjugation divisions and keeping track of the age of the clone. PMID- 8719874 TI - A 182 bp fragment of the mouse pro alpha 1(II) collagen gene is sufficient to direct chondrocyte expression in transgenic mice. AB - Type II collagen is a major chondrocyte-specific component of the cartilage extracellular matrix and it represents a typical differentiation marker of mature chondrocytes. In order to delineate cis-acting elements of the mouse pro alpha 1(II) collagen gene that control chondrocyte-specific expression in intact mouse embryos, we generated transgenic mice harboring chimeric constructions in which varying lengths of the promoter and intron 1 sequences were linked to a beta galactosidase reporter gene. A construction containing a 3,000 bp promoter and a 3,020 bp intron 1 fragment directed high levels of beta-galactosidase expression specifically to chondrocytes. Expression of the transgene coincided with the temporal expression of the endogenous gene at all stages of embryonic development. Successive deletions of intron 1 delineated a 182 bp fragment which targeted beta-galactosidase expression to chondrocytes with the same specificity as the larger intron 1 fragment. Transgenic mice harboring a 309 bp Col2a1 promoter lacking intron 1 tester sequences showed no beta-galactosidase expression in chondrocytes. Reduction of the 182 bp fragment to a 73 bp subfragment surrounding a decamer sequence previously reported to be involved in chondrocyte specificity, resulted in loss of transgene expression in chondrocytes. When the Col2a1 promoter was replaced with a minimal beta-globin promoter, the 182 bp intron 1 sequence was still able to target expression of the transgene to chondrocytes. We conclude that a 182 bp intron 1 DNA segment of the mouse Col2a1 gene contains the necessary information to confer high-level, temporally correct, chondrocyte expression on a reporter gene in intact mouse embryos and that Col2a1 promoter sequences are dispensable for chondrocyte expression. PMID- 8719875 TI - Matrigel induces thymosin beta 4 gene in differentiating endothelial cells. AB - We performed differential cDNA hybridization using RNA from endothelial cells cultured for 4 hours on either plastic or basement membrane matrix (Matrigel), and identified early genes induced during the morphological differentiation into capillary-like tubes. The mRNA for one clone, thymosin beta 4, was increased 5 fold. Immunostaining localized thymosin beta 4 in vivo in both growing and mature vessels as well as in other tissues. Endothelial cells transfected with thymosin beta 4 showed an increased rate of attachment and spreading on matrix components, and an accelerated rate of tube formation on Matrigel. An antisense oligo to thymosin beta 4 inhibited tube formation on Matrigel. The results suggest that thymosin beta 4 is induced and likely involved in differentiating endothelial cells. Thymosin beta 4 may play a role in vessel formation in vivo. PMID- 8719876 TI - Lung surfactant protein A (SP-A) activates a phosphoinositide/calcium signaling pathway in alveolar macrophages. AB - Lung surfactant protein A (SP-A), the main protein component of lung surfactant which lines the alveoli, strongly enhances serum-independent phagocytosis of bacteria by rat alveolar macrophages. We tested if the effect of SP-A is due to interaction with the macrophages or to opsonization of the bacteria. In phagocytosis assays with fluorescein isothiocyanate labeled bacteria, SP-A had no opsonic effect on Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus, but enhanced phagocytosis by acting only on the macrophages. We characterized this activation mechanism. With single cell measurements of fura-2 loaded cells we demonstrate that SP-A raises the intracellular free calcium ion concentration 6 to 8 seconds after addition. This calcium mobilization is dose dependent in that increased SP-A concentrations lead to a higher percentage of responding cells. Additionally, SP-A leads to a dose-dependent and transient generation of inositol 1,4.5-trisphosphate. Release of intracellular stored calcium by SP-A is a prerequisite for its stimulatory effect on phagocytosis, since SP-A-induced enhancement of phagocytosis can be impaired by prior addition of thapsigargin, a Ca(2+)-ATPase inhibitor that leads to depletion of intracellular calcium stores. We conclude that SP-A activates a phosphoinositide/calcium signaling pathway in alveolar macrophages leading to enhanced serum-independent phagocytosis of bacteria. PMID- 8719877 TI - The hepatocyte growth factor/scatter factor (HGF/SF) receptor, met, transduces a morphogenetic signal in renal glomerular fibromuscular mesangial cells. AB - Previous studies have demonstrated that hepatocyte growth factor/scatter factor (HGF/SF) is secreted by mesenchymal cells and that it elicits motility, morphogenesis and proliferation of epithelia expressing the met receptor. We now report that HGF/SF may act as an autocrine factor in fibromuscular renal mesangial cells. These cells mechanically support glomerular endothelia, control the rate of plasma ultrafiltration and are implicated in the pathogenesis of a variety of chronic renal diseases. We detected met protein in the vascular stalk of metanephric glomeruli and in the mature mesangium. Mesangial lines from a mouse transgenic for a temperature-sensitive simian virus 40 T antigen expressed met mRNA and protein, and recombinant HGF/SF phosphorylated the met receptor tyrosine kinase. Cells were immortal in the permissive condition and HGF/SF enhanced proliferation in a defined medium. In the absence of the immortalising protein, division ceased and recombinant HGF/SF caused multipolar cells to become bipolar. The factor diminished stress fibres, their focal contacts and immunostaining for extracellular fibronectin, hence suggesting reduced substratum adhesion and enhanced motility. Mesangial lines also expressed HGF/SF mRNA and secreted bioactive factor; immunocytochemistry showed both ligand and receptor in individual cells. HGF/SF blocking antibody aggregated the cells, suggesting that mesangial-derived factor affects basal cell conformation in an autocrine manner. We conclude that mesangial cells express both HGF/SF and met, and the factor induces morphogenesis of cultured mesangial cells. Therefore HGF/SF may have an autocrine role in mesangial biology but further studies are now required to investigate the potential importance of the factor in vivo. PMID- 8719878 TI - The role of macrophage receptors in adhesion and uptake of Leishmania mexicana amastigotes. AB - Amastigotes of the protozoan parasite Leishmania proliferate in phagolysosomes of mammalian macrophages. Propagation of the infection is considered to occur by host-cell rupture and uptake of released parasites by uninfected macrophages. In this study, the kinetics of binding of L mexicana mexicana amastigotes to COS cells and to COS cells transfected with three different macrophage receptors (FcRII-B2, receptor for the Fc-domain of immunoglobulins; CR3, complement type 3 receptor and the mannose receptor) is compared to the rate of adhesion to peritoneal macrophages. Amastigotes isolated from macrophages cultivated in vitro bind with slow, sigmoid kinetics to COS cells expressing either of the three receptors, or to peritoneal macrophages. In contrast, amastigotes isolated from mouse lesions bind with rapid, hyperbolic kinetics to COS cells expressing the Fc receptor or to peritoneal macrophages but with slow, sigmoid kinetics to COS cells expressing the CR3 or the mannose receptor. As shown by immunofluorescence experiments, lesion-derived amastigotes contain host-derived immunoglobulins (Ig) but no complement component 3 at their surface. It is concluded that amastigotes contain no intrinsic ligand at their surface, which enables high-affinity interactions with macrophages. Opsonization by specific Ig may be of relevance in vivo because firstly, in cryosections of mouse lesions extracellular amastigotes containing surface Ig can be detected and, secondly, B cell-deficient mice reconstituted with parasite-specific Ig show a modest increase in the rate of lesion development. In addition, it is shown that amastigotes are internalized by COS cells and grow in large parasitophorous vacuoles similar to those observed in macrophages. PMID- 8719879 TI - Synthesis and assembly of human beta 1 gap junctions in BHK cells by DNA transfection with the human beta 1 cDNA. AB - Gap junctional communication is important in many physiological processes, including growth control, patterning, and the synchronization of cell-to-cell activities. It has been difficult to study the synthesis and assembly of gap junctions due to their low abundance. To overcome this limitation, baby hamster kidney cells (BHK) have been transfected with a human beta 1 (Cx32) connexin cDNA construct. Expression was placed under the control of the mouse metallothionein promoter that can be induced by heavy metals. The transfected cells were characterized by DNA, RNA and protein analysis, as well as by scrape loading to detect functional channels. Functional beta 1 connexin was detected only in cells transfected with beta 1 connexin cDNA in the correct orientation (beta 1-BHK). Analysis of the cells by light microscopic immunocytochemistry indicated that beta 1 connexin antigen was localized to the plasma membrane and to several intracellular compartments. Characterization with thin section electron microscopy revealed extensive areas of assembled double membrane gap junctions between cells (on the cell surface), in the endoplasmic reticulum (ER), and the nuclear envelope. This unusual intracellular distribution for assembled gap junction protein was confirmed by freeze fracture analysis, which revealed large particle aggregates, characteristic of gap junction plaques, on the fracture faces of all these membranes. The presence of gap junction particle aggregates in the ER suggests that the oligomerization of connexin can occur at its site of synthesis. Further, the process of assembly into double membrane junction structures in intracellular membranes may be driven by connexin protein concentration. PMID- 8719880 TI - A tumor-associated glycosylation change in the glucose transporter GLUT1 controlled by tumor suppressor function in human cell hybrids. AB - Studies of human cell hybrids have provided evidence that the tumorigenicity of a cervical carcinoma (HeLa) is under the control of a putative tumor suppressor on chromosome 11. Using these human cell hybrids, we found a tumor-associated glycosylation change in the glucose transporter GLUT1, which is an N-linked glycoprotein at the plasma membrane. The non-tumorigenic HeLa x fibroblast cell hybrid CGL1 and the normal diploid fibroblast WI38 expressed the 50-55 kDa GLUT1, whereas in a tumorigenic segregant hybrid, CGL4, as well as in parental HeLa cells, GLUT1 glycosylation was altered and its molecular mass was about 70 kDa. However, the altered GLUT1 glycosylation was not observed in SV40-transformed WI38 cells, suggesting a correlation between this glycosylation change and a putative tumor suppressor function. Further investigations using glycosidases, glycosylation inhibitors and lectin-affinity chromatography demonstrated that the tumor-associated glycosylation change in GLUT1 was mainly due to the increase in N-acetyl-lactosamine repeats in the N-linked oligosaccharides. In accordance with the altered glycosylation, affinity for 2-deoxyglucose in the tumorigenic CGL4 cells increased 2-fold, but there was little change in the Vmax. These results suggest there may be a functional role for the modulation by glycosylation of GLUT1 in the tumorigenic behavior of CGL4 and HeLa cells. PMID- 8719881 TI - Schizosaccharomyces pombe Vps34p, a phosphatidylinositol-specific PI 3-kinase essential for normal cell growth and vacuole morphology. AB - We have cloned the gene, vps34+, from the fission yeast Schizosaccharomyces pombe which encodes an 801 amino acid protein with phosphatidylinositol 3-kinase activity. The S. pombe Vps34 protein shares 43% amino acid sequence identity with the Saccharomyces cerevisiae Vps34 protein and 28% identity with the p110 catalytic subunit of the mammalian phosphatidylinositol 3-kinase. When the vps34+ gene is disrupted, S.pombe strains are temperature-sensitive for growth and the mutant cells contain enlarged vacuoles. Furthermore, while wild-type strains exhibit substantial levels of phosphatidylinositol 3-kinase activity, this activity is not detected in the vps34 delta strain. S.pombe Vps34p-specific antiserum detects a single protein in cells of -90 kDa that fractionates almost exclusively with the crude membrane fraction. Phosphatidylinositol 3-kinase activity also is localized mainly in the membrane fraction of wild-type cells. Immunoisolated Vps34p specifically phosphorylates phosphatidylinositol on the D-3 position of the inositol ring to yield phosphatidylinositol(3)phosphate. but does not utilize phosphatidylinositol(4)phosphate or phosphatidylinositol(4,5)bisphosphate as substrates. In addition, when compared to the mammalian p110 phosphatidylinositol 3-kinase, S. pombe Vps34p is relatively insensitive to the inhibitors wortmannin and LY294002. Together, these results indicate that S. pombe Vps34 is more similar to the phosphatidylinositol specific 3-kinase, Vps34p from S. cerevisiae, and is distinct from the p110/p85 and G protein-coupled phosphatidylinositol 3-kinases from mammalian cells. These data are discussed in relation to the possible role of Vps34p in vesicle-mediated protein sorting to the S. pombe vacuole. PMID- 8719882 TI - Identification of a Ca(2+)-binding light chain within Chlamydomonas outer arm dynein. AB - We describe here the molecular cloning of the M(r) 18,000 dynein light chain from the outer arm of Chlamydomonas flagella. In vivo, this molecule is directly associated with the gamma dynein heavy chain. Sequence analysis indicates that this light chain is a novel member of the calmodulin superfamily of Ca2+ binding regulatory proteins; this molecule is 42, 37 and 36% identical to calmodulin, centrin/caltractin and troponin C, respectively, and also shows significant similarity to myosin light chains. Although four helix-loop-helix elements are evident, only two conform precisely to the EF hand consensus and are therefore predicted to bind Ca2+ in vivo. In vitro Ca2+ binding studies indicate that this dynein light chain (expressed as a C-terminal fusion with maltose binding protein) has at least one functional Ca2+ binding site with an apparent affinity for Ca2+ of approximately 3 x 10(-5) M. Within the Chlamydomonas flagellum, the transition from an assymmetric to a symmetric waveform (which implies an alteration in dynein activity) is mediated by an increase in intraflagellar Ca2+ from 10(-6) to 10(-1) M; this transition is altered in mutants that lack the outer arm. The data presented here suggest that a Ca(2+)-dependent alteration in the interaction of this dynein light chain with the motor containing heavy chain may affect outer arm function during flagellar reversal. PMID- 8719883 TI - Protocadherin Pcdh2 shows properties similar to, but distinct from, those of classical cadherins. AB - Cell adhesion and several other properties of a recently identified cadherin related protein, protocadherin Pcdh2, were characterized. A chimeric Pcdh2 in which the original cytoplasmic domain was replaced with the cytoplasmic domain of E-cadherin was expressed in mouse L cells. The expressed protein had a molecular mass of about 150 kDa and was localized predominantly at the cell periphery, as was the wild-type Pcdh2. In a conventional cell aggregation assay, the transfectants showed cell aggregation activity comparable to that of classical cadherins. This activity was Ca(2+)-dependent and was inhibited by the addition of anti-Pcdh2 antibody, indicating that the chimeric Pcdh2, and probably the wild type Pcdh2, has Ca(2+)-dependent cell aggregation activity. Mixed cell aggregation assay using L cells and different types of transfectants showed that the activity of Pcdh2 was homophilic and molecular type specific and that Pcdh2 was transfectants did not aggregate with other types of transfectants or with L cells. In immunoprecipitation, the chimeric Pcdh2 co-precipitated with a 105 kDa and a 95 kDa protein, whereas wild-type Pcdh2 co-precipitated with no major protein. Pcdh2 was easily solubilized with non-ionic detergent, in contrast to the case of classical cadherins. On immunofluorescence microscopy, the somas of Purkinje cells were diffusely stained with anti-human Pcdh2 antibody. Mouse Pcdh1 and Pcdh2 were mapped to a small segment of chromosome 18, suggesting that various protocadherins form a gene cluster at this region. The present results suggest that Pcdh2, and possibly other protocadherins as well as protocadherin related proteins such as Drosophila fat, mediate Ca(2+)-dependent and specific homophilic cell-cell interaction in vivo and play an important role in cell adhesion, cell recognition, and/or some other basic cell processes. PMID- 8719884 TI - Localization of the rat myosin I molecules myr 1 and myr 2 and in vivo targeting of their tail domains. AB - Myr 1 is a widely distributed mammalian myosin I molecule related to brush border myosin 1. A second widely distributed myosin I molecule similar to myr 1 and brush border myosin I, called myr 2, has now been identified. Specific antibodies and expression of epitope-tagged molecules were used to determine the subcellular localization of myr 1 and myr 2 in NRK cells. Myr 1 was detected at the plasma membrane and was particularly enriched in cell protrusions like lamellipodia, membrane ruffles and filopodia. In dividing cells myr 1 localized to the cleavage furrow. Myr 2 was localized in a discrete punctate pattern in resting cells and in cells undergoing cytokinesis. In subcellular fractionation experiments myr 1 and myr 2 were both partly soluble and partly associated with smooth membranes of medium density. The tail domains of myosin I molecules have been proposed to interact with a receptor and thereby determine the subcellular localization. To test this hypothesis we expressed the tail domains of myr 1 and myr 2 that lack the F-actin-binding myosin head domain in NRK cells. These tail domains also partly copurified with smooth membranes of medium density and immunolocalized similar to the respective endogenous myosin I; however, they exhibited a lower affinity for membranes and an increased diffuse cytosolic localization. These results suggest that the tail domains of myr 1 and myr 2 are sufficient for subcellular targeting but that their head domains also contribute significantly to maintaining a proper subcellular localization. PMID- 8719885 TI - Cell spreading and the regulation of ornithine decarboxylase. AB - The aim of this study was to investigate the effect of cell spreading on the induction of ornithine decarboxylase and the rate of putrescine uptake in anchorage-dependent and anchorage-independent cells. Plating non-transformed IEC 6 epithelial cells at high versus low cell density restricted cell spreading from 900 microns 2 to approximately 140 microns 2, blunted the transient induction of ornithine decarboxylase activity from 202 to 32 pmol 14CO2/mg protein per hour and reduced the rate of [14C] putrescine uptake from 46 to 23 pmol/10(5) cells per hour. The mean spreading area of the cell population was controlled by coating tissue culture dishes with the nonadhesive polymer, polyHEMA. Ornithine decarboxylase activity and putrescine uptake correlated with cell spreading with minimal spreading (263 microns 2) corresponding to an 83% decrease in ornithine decarboxylase activity and 51% decrease in the rate of putrescine uptake. Adding the RGD peptide, Gly-Arg-Gly-Glu-Ser-Pro to the medium of sparsely plated cells resulted in rapid reductions in cell spreading concomitant with dose-dependent decreases in ornithine decarboxylase activity and putrescine uptake. Finally, minimizing cell spreading by depriving cells of substratum contact completely abolished serum-induced increases in ornithine decarboxylase and reduced the rate of putrescine uptake by 47%. In contrast to IEC-6 cells, ornithine decarboxylase of neoplastic HTC-116 cells was constitutively expressed with basal and stimulated activity (193 and 982 pmol 14CO2/mg protein per hour, respectively) completely independent of cell adhesion. Putrescine uptake, however, was abolished in the absence of cell adhesion. These data suggest that the induction of ornithine decarboxylase activity and the rate of putrescine uptake correlate with spreading of anchorage-dependent IEC-6 cells and that ornithine decarboxylase activity but not putrescine uptake, appears to be independent of spreading of neoplastic HTC-116 cells. PMID- 8719886 TI - Expression of laminin isoforms in mouse myogenic cells in vitro and in vivo. AB - The expression of laminin-1 (previously EHS laminin) and laminin-2 (previously merosin) isoforms by myogenic cells was examined in vitro and in vivo. No laminin alpha 2 chainspecific antibodies react with mouse tissues, 50 rat monoclonal antibodies were raised against the mouse laminin alpha 2 chain: their characterization is described here. Myoblasts and myotubes from myogenic cell lines and primary myogenic cultures express laminin beta 1 and gamma 1 chains and form a complex with a 380 kDa alpha chain identified as laminin alpha 2 by immunofluorescence, immunoprecipitation and PCR. PCR from C2C12 myoblasts and myotubes for the laminin alpha 2 chain gene (LamA2) provided cDNA sequences which were used to investigate the in vivo expression of mouse LamA2 mRNA in embryonic tissues by in situ hybridization. Comparisons were made with specific probes for the laminin alpha 1 chain gene (LamA1). LamA2 but not LamA1 mRNA was expressed in myogenic tissues of 14- and 17-day-old mouse embryos, while the laminin alpha 2 polypeptide was localized in adjacent basement membranes in the muscle fibres. In situ hybridization also revealed strong expression of the LamA2 mRNA in the dermis, indicating that laminin alpha 2 is expressed other than by myogenic cells in vivo. Immunofluorescence studies localized laminin alpha 2 in basement membranes of basal keratinocytes and the epithelial cells of hair follicles, providing new insight into basement membrane assembly during embryogenesis. In vitro cell attachment assays revealed that C2C12 and primary myoblasts adhere to laminin-1 and -2 isoforms in a similar manner except that myoblast spreading was significantly faster on laminin-2. Taken together, the data suggest that laminins 1 and 2 play distinct roles in myogenesis. PMID- 8719887 TI - Receptor-mediated adhesive and anti-adhesive functions of chondroitin sulfate proteoglycan preparations from embryonic chicken brain. AB - Chondroitin sulfate proteoglycans inhibit the adhesion of cells to extracellular matrix proteins that otherwise permit adhesion. Although proteoglycans are widely assumed to act by masking the other protein in a mixed substrate, recent studies suggest that proteoglycans inhibit adhesion through mechanisms initiated by their binding to specific cell surface receptors. To explore this issue, we developed a purification scheme to isolate proteoglycan aggregates, monomers, and core proteins. Two distinct adhesion assays were used to study the interaction of these proteoglycan preparations with human foreskin fibroblasts: the gravity assay in which cell attachment is stabilized by cell spreading, and the centrifugation assay in which spreading does not play a role. All proteoglycan preparations mediate adhesion in the centrifugation assay but not in the gravity assay. In the centrifugation assay, proteoglycan aggregates and monomers are considerably more active than other extracellular matrix proteins while proteoglycan core proteins are at least as active as other extracellular matrix proteins. Proteoglycan core proteins bind to cell-associated hyaluronic acid, but not to integrins. Using mixed substrates in the gravity assay, all proteoglycan preparations inhibited cell attachment to fibronectin and vitronectin but not to collagen I and laminin. Although proteoglycan aggregates and monomers are more active than core proteins in inhibiting adhesion in the gravity assay, core proteins are still clearly active. A variety of control experiments suggest that the inhibition of cell attachment by proteoglycans is mediated through the specific interactions of proteoglycans with cell surface receptors, resulting in the inhibition of cell spreading. These results suggest at least two molecular mechanisms for proteoglycan-fibroblast interactions, one involving the chondroitin sulfate on the proteoglycan and an as yet unidentified receptor, the other involving the proteoglycan core protein and cell-associated hyaluronic acid. PMID- 8719888 TI - Integrin signaling: roles for the cytoplasmic tails of alpha IIb beta 3 in the tyrosine phosphorylation of pp125FAK. AB - pp125FAK (focal adhesion kinase) a protein tyrosine kinase that may mediate cellular responses to adhesion, is activated and tyrosine-phosphorylated when platelets adhere to fibrinogen via the integrin, alpha IIb beta 3. To determine whether either of the cytoplasmic tails of alpha IIb beta 3 regulates FAK phosphorylation, CHO cells were stably transfected with alpha IIb beta 3 or various cytoplasmic tail truncation mutants. Cells expressing wild-type alpha IIb beta 3 or alpha IIb beta 3 that lacked the COOH-terminal 13 or 18 residues of the 20 residue alpha IIb tail adhered to and spread on fibrinogen or on an anti-alpha IIb antibody, and FAK became tyrosine-phosphorylated. FAK also became phosphorylated in adherent cells lacking the COOH-terminal 35 or 39 residues of the 47 residue beta 3 tail, although the extent of phosphorylation was reduced by about 50% in the latter mutant. Little or no FAK phosphorylation was observed if 46 residues were deleted from the beta 3 tail. None of these beta 3 truncation mutants spread on the anti-alpha IIb antibody. When cells with wild-type alpha IIb beta 3 or truncated beta 3 were detached from a surface, FAK became rapidly dephosphorylated. In contrast, FAK remained phosphorylated in the two alpha IIb truncation mutants for up to 90 minutes in suspension. This persistent phosphorylation was not due to occupancy of alpha IIb beta 3 by adhesive ligands because it was also observed with an alpha IIb tail truncation mutant that contained an additional mutation in the extracellular portion of the receptor that prevents ligand binding. These studies demonstrate that: (1) the beta 3 cytoplasmic tail, including the membrane-proximal portion, is involved in initiation of FAK phosphorylation; (2) FAK phosphorylation can be initiated by cell adhesion in the absence of cell spreading; and (3) the membrane-distal portion of the alpha IIb cytoplasmic tail may normally function to dampen FAK phosphorylation in non-anchored cells. PMID- 8719889 TI - Differential targeting of recombinant fibronectins in AtT-20 cells based on their efficiency of aggregation. AB - In pituitary-derived AtT-20 cells, recombinant fibronectin containing the N terminal matrix assembly domain and the C-terminal half of fibronectin does not follow the regulated secretory pathway but instead concentrates in distinct organelles prior to secretion. These organelles are larger than the dense-core granules and localize to the cell body at sites that differ from lysosomes, endosomes and endoplasmic reticulum. Unlike the dense-core granules, their discharge is not stimulated by 8-bromo-cyclic-AMP or phorbol esters. The kinetics of intracellular transport and secretion of the recombinant fibronectin suggest that it is present in a post-Golgi pool that turns over more slowly than constitutive vesicles. Indeed, the fibronectin-containing organelles disappear with a half-time of 3 hours after inhibiting protein synthesis. Presence of the organelles correlates with intracellular aggregation of dimeric fibronectin polypeptides. The organelles are absent in cells expressing monomeric recombinant fibronectin (lacking C-terminal dimerization sites) or the C-terminal half of fibronectin (which dimerizes but lacks the N-terminal matrix assembly domain), both of which aggregate less efficiently than dimeric fibronectin. Instead, the latter polypeptides enter the dense-core granules. Thus while the formation of the fibronectin-containing organelles may require efficient aggregation, it may not require a specific structural signal. Moreover, efficient aggregation is not necessarily a prerequisite for following the regulated pathway. PMID- 8719890 TI - Control of N-cadherin-mediated intercellular adhesion in migrating neural crest cells in vitro. AB - Dispersion of neural crest cells and their ultimate regroupment into peripheral ganglia are associated with precisely coordinated regulations both in time and space of the expression and function of cell adhesion receptors. In particular, the disappearance of N-cadherin from the cell surface at the onset of migration and its reexpression during cell aggregation suggest that, during migration, N cadherin expression is repressed in neural crest cells. In the present study, we have analyzed in vitro the mechanism of control of N-cadherin expression and function in migrating neural crest cells. Although these cells moved as a dense population, each individual did not establish extensive and permanent intercellular contacts with its neighbors. However, cells synthesized and expressed mature N-cadherin molecules at levels comparable to those found in cells that exhibit stable intercellular contacts, but in contrast to them, the bulk of N-cadherin molecules was not connected with the cytoskeleton. We next determined which intracellular events are responsible for the instability of the N-cadherin junctions in neural crest cells using various chemical agents known to affect signal transduction processes. Agents that block a broad spectrum of serine-threonine kinases (6-dimethylaminopurine, H7 and staurosporine) or that affect selectively protein kinases C (bisindolylmaleimide and sphingosine), inhibitors of protein tyrosine kinases (erbstatin, herbimycin A, and tyrphostins), and inhibitors of phosphatases (vanadate) all restored tight cell cell associations among neural crest cells, accompanied by a slight increase in the overall cellular content of N-cadherin and its accumulation to the regions of intercellular contacts. The effect of the kinase and phosphatase blockers was inhibitable by agents known to affect protein synthesis (cycloheximide) and exportation (brefeldin A), indicating that the restored cell-cell contacts were mediated chiefly by an intracellular pool of N-cadherin molecules recruited to the membrane. Finally, N-cadherin molecules were constitutively phosphorylated in migrating neural crest cells, but their level and state of phosphorylation were apparently not modified in the presence of kinase and phosphatase inhibitors. These observations therefore suggest that N-cadherin-mediated cell-cell interactions are not stable in neural crest cells migrating in vitro, and that they are under the control of a complex cascade of intracellular signals involving kinases and phosphatases and probably elicited by surface receptors. PMID- 8719891 TI - Microtubule organization and the distribution of gamma-tubulin in spermatogenesis of a beetle, Tenebrio molitor (Tenebrionidae, Coleoptera, Insecta). AB - The present study focuses on the restructuring of the microtubule (MT) cytoskeleton and microtubule-organizing centres (MTOCs) throughout spermatogenesis of a darkling beetle, Tenebrio molitor (Tenebrionidae, Coleoptera, Insecta). To this end, serial ultrathin sections through male germ cells were studied using transmission electron microscopy. Additionally, spindles and young spermatids were isolated from testes under MT-stabilizing conditions and doubly labeled with antibodies against beta- and gamma-tubulin. The latter is a tubulin isoform detected in MTOCs of a wide variety of species. The observations suggest that microtubules may be nucleated from sites with and without high gamma-tubulin content and that these sites do not necessarily possess canonical centrosomes. In a prominent cytoplasmic MT system of primary spermatocytes in prophase, microtubule nucleation apparently occurs in the absence of immunologically detectable gamma-tubulin. At the poles of meiotic spindles, MTs are directly inserted into gamma-tubulin-containing material and this connection is considered responsible for their nucleation. The interzone spindle MTs of telophase cells contain gamma-tubulin and this may confer stability to them. Finally, manchette MTs of spermatids originate in the vicinity of the acrosome precursor but are not inserted into this body. The acrosome precursor is surrounded by a membrane and is clearly detected by the antibody against gamma-tubulin. PMID- 8719892 TI - Message of nexin 1, a serine protease inhibitor, is accumulated in the follicular papilla during anagen of the hair cycle. AB - A group of specialized mesenchymal cells located at the root of the mammalian hair follicle, known as the follicular or dermal papillary cells, are involved in regulating the hair cycle, during which keratinocytes of the lower follicle undergo proliferation, degeneration and regrowth. Using the arbitrarily primed PCR approach, we have identified a 1.3 kb messenger RNA that is present in large quantities in cultured rat follicular papillary cells, but not in skin fibroblasts. This mRNA encodes nexin 1, a potent protease inhibitor that can inactivate several growth-modulating serine proteases including thrombin, urokinase and tissue plasminogen activator. In situ hybridization showed that nexin 1 message is accumulated in the follicular papilla cells of anagen follicles, but is undetectable in keratinocytes or other skin mesenchymal cells. In addition, nexin 1 message level varies widely among several immortalized rat vibrissa papillary cell lines, and these levels correlate well with the reported abilities of these cell lines to support in vivo follicular reconstitution. These results suggest a possible role of nexin 1 in regulating hair follicular growth. PMID- 8719893 TI - Low serum erythropoietin--a strong diagnostic criterion of primary polycythaemia even at normal haemoglobin levels. AB - Estimations of serum erythropoietin level by enzyme immunoassay (EIA) kit were made in 42 patients with primary polycythaemia, and in a comparison group consisting of 41 patients with secondary polycythaemia and 47 patients with idiopathic erythrocytosis. The majority of patients with primary polycythaemia were undergoing treatment by venesection and therefore had Hb levels in the normal range at the time of erythropoietin estimation. In primary polycythaemia, 64% of the first samples taken from each patient were below the reference range for serum erythropoietin in normal individuals. When two samples were taken from each patient 72% had low values in one or both samples. In the comparison group, analysis of those patients who had two samples taken showed only one individual with secondary polycythaemia and none with idiopathic erythrocytosis who had a serum erythropoietin level below the reference range. The finding of low serum erythropoietin in patients with primary polycythaemia, even when Hb levels are normal due to venesection, is of high diagnostic specificity (few false positive results) and useful diagnostic sensitivity (28% false negative results). It is proposed that future diagnostic criteria of primary polycythaemia should include the finding of a serum erythropoietin level below the lower limit of normal in at least one of two serum samples taken on different occasions. PMID- 8719894 TI - Enzyme-linked antiglobulin test for the evaluation of the amount of IgG autoantibodies on red blood cells. AB - An enzyme-linked antiglobulin test (ELAT) was adapted to the assessment of IgG autoantibodies on red cells. Due to the worked out standard curve and the performance of the test on microplates, ELAT became a simple and inexpensive technique. It can be used for the detection of IgG within the range from 70 to 37,500 molecules per red cell. This is a simple quantitative method that can be used in immunohaematological studies of red cells. PMID- 8719895 TI - Evaluation of monocyte counting with two automated instruments by the use of CD 14-specific immunomagnetic Dynabeads. AB - Accurate determination of monocytes can be a problem with automated haematological analysers as well as with visual microscopy. We have compared monocyte counts from our two routine automated instruments, Technicon H*1 and Sysmex NE-1500. In the NE-1500 the monocyte counts appear to increase with increasing concentrations of neutrophils, compared with the H*1 results. The accuracy of the monocyte counts was assessed using Dynabeads M-450 CD14. These uniform magnetizable polystyrene beads coated with monoclonal antibody specific for CD14 membrane antigen isolated monocytes from the blood samples. The influence of this depletion on the monocyte counts and histograms from the two instruments and by visual microscopy were compared. On average 90% of the monocytes counted by visual microscopy were removed from six blood samples. In the same samples the beads removed an average of 75% of cells identified as monocytes by the H*1 but only 49% of those identified as monocytes by the NE 1500. These Dynabeads can be a useful tool in evaluating monocyte identification in different haematological situations and for manufacturers developing technology for automated leucocyte differential counting. PMID- 8719896 TI - Recombinant thromboplastin inhibition assay for the detection of lupus anticoagulant. AB - Tissue thromboplastin inhibition assay (TTI) is a sensitive test for lupus anticoagulant (LA). We have performed TTI in 12 LA positive patients using a new recombinant human tissue factor (Innovin, IN) and compared it with Thromborel S (TH), a commonly used human placenta thromboplastin. The effect of using two different dilutions of each thromboplastin (1:10 & 1:26) was investigated. A 1:26 dilution discriminated better than the 1:10 and this was more evident for Innovin. The mean value obtained with a 1:26 IN dilution was statistically different from that observed with TH at the same dilution. Furthermore, when PT and TTI ratios were considered, differences were statistically significant and seemed to increase depending on thromboplastin dilutions. When we used IN at 1:26 all LA positive patients had a value > 1.2. Then we compared TTI at a 1:26 dilution with dilute Russell's Viper Venom Time (dRVVT) in 50 consecutive patients with suspected lupus anticoagulant not treated with warfarin or heparin. In these patients the diagnosis of lupus anticoagulant was carried out using dilute APTT mixing studies and a platelet neutralization procedure: four out of 50 patients thus tested were LA positive. When dRVVT or TTI-I 1:26 were used, five patients were positive for lupus anticoagulant. Innovin showed similar sensitivity of dRVVT for detection of lupus anticoagulant. It is likely that higher dilutions of thromboplastins could further improve the specificity of this method. PMID- 8719897 TI - Type III procollagen N-peptide correlates with beta-2-microglobulin in myelomatosis. AB - The serum concentration of the N-terminal peptide of type III procollagen (PIIINP) was determined in 32 patients with myelomatosis (MM). Four subjects were studied at the time of diagnosis and the remaining patients at variable intervals from diagnosis. Serum concentration of beta-2-microglobulin (B2m) was measured in 31 patients. Serial measurements of both substances were performed in 20 patients. Serum PIIINP was increased in MM as compared with healthy control subjects (P < 0.001). Patients with active disease had significantly higher propeptide values (median 7.4; range 3.8-11.2) as compared to those with stable disease (median 4.3; range 2.2-9.6) (P < 0.009). A highly significant correlation existed between circulating PIIINP and B2m (P < 0.001). It is concluded that MM elicits a stromal reaction as reflected by parallel increases in serum PIIINP and serum B2m. In subsets of patients, e.g. those with non-secretory myeloma and in patients with smouldering disease, serum PIIINP may even be superior to B2m as an indicator of disease activity. PMID- 8719898 TI - Acute non-lymphoblastic leukaemia with t(16;21): case report with a review of the literature. AB - A rare case of acute non-lymphoblastic leukaemia with chromosomal t(16;21)(p11;q22) translocation was studied at the molecular level. Most of the previous reports about the translocation have been described at a karyotypic level. Blast cells of this patient expressed the TLS-ERG chimeric mRNA. The clinical, morphological, karyotypic, and immunohistochemical aspects of this leukaemia are also presented with a review of the literature. PMID- 8719899 TI - An unusual case of hypersomnolence syndrome. AB - Hypersomnolence syndrome has been well described in patients with acute lymphoblastic leukaemia (ALL) following cranial irradiation. It is most frequently seen in children. In adults it is clinically very mild. We describe a case of a particularly severe and prolonged post radiation hypersomnolence syndrome in an adult with ALL. The hypersomnolence syndrome is discussed in the context of side effects of central nervous system directed therapy in elderly patients with ALL. PMID- 8719900 TI - Letter from America: the end of civilization as we know it? PMID- 8719901 TI - Further comparative experience with the Roche Cobas Argos Analyser. PMID- 8719902 TI - Haematology self-assessment exercise. PMID- 8719903 TI - Prediction of urinary sulphate and glucuronide conjugate excretion for substituted phenols in the rat using quantitative structure-metabolism relationships. AB - 1. The quantitative urinary excretion of the sulphate and glucuronide metabolites of 15 substituted phenols dosed to rat has been determined using high resolution 19F-nmr spectroscopy. 2. The urinary metabolic fate of each of the compounds was related to a series of calculated physicochemical properties for each compound to produce quantitative structure-metabolism relationships (QSMRs). Using these calculated molecular properties it was possible to predict the urinary recovery of xenobiotic material as a percentage of the administered dose, to classify the compounds according to their 'dominant' metabolite pattern and to predict quantitatively the proportions of glucuronide and sulphate conjugates in the urine by the use of multiple linear regression. 3. The quantitative predictions were tested by cross-validation and good prediction of total xenobiotic urinary recovery as a percentage of the administered dose was achieved based on an equation involving the electrophilic superdelocalizability at C4 (para to the hydroxyl function), the smallest principal ellipsoid axis dimension and the heat of formation. The largest moment of inertia and the electrophilic superdelocalizability at C3 were found to be the most significant factors for the prediction of the percentage glucuronide in the urine, and the urinary excretion of sulphate conjugates as a percentage of total urinary recovery was negatively correlated with the glucuronide excretion as little parent compound was excreted. PMID- 8719904 TI - S-oxidation of (+)-cis-3,5-dimethyl-2-(3-pyridyl)-thiazolidin-4-one hydrochloride by rat hepatic flavin-containing monooxygenase 1 expressed in yeast. AB - 1. Rat hepatic flavin-containing monooxygenase 1 (FMO1) expressed in yeast catalyzed the S-oxidation of (+)-cis-3,5-dimethyl-2-(3-pyridyl)thiazolidin-4-one hydrochloride (SM-12502) in vitro. 2. S-oxidation was inhibited by 1-(1-naphthyl) 2-thiourea and thiobenzamide, known inhibitors of FMO, but was not enhanced by n octylamine, a known enhancer of FMO. 3. The rate of S-oxide formation from SM 12502 was about four-fold lower than that from (+/-)-trans-3,5-dimethyl-2-(3 pyridyl)thiazolidin-4-one hydrochloride (SM-9979) and enantioselectivity and diastereoselectivity of the S-oxidation reaction were observed. 4. The ability of the recombinant yeast to produce the S-oxide from SM-12502 was maintained for long periods and exemplifies the recombinant yeast as a bioreactor to produce a large amount of the S-oxide. PMID- 8719905 TI - Saturable binding sites for the coenzyme A ester of nafenopin, a peroxisome proliferator, in rat liver cytosol. AB - 1. At least three different molecular weight binding sites exist in rat liver cytosol for nafenopin-CoA, the coenzyme A ester and metabolic product of the carcinogenic peroxisome proliferator nafenopin. No binding sites for the free drug were observed. 2. Polypeptides of 35-40 kDa molecular weight range where no acyl-CoA binding proteins have been previously described bind the highest proportion of nafenopin-CoA (60-70%). Binding is displaceable by the CoA esters of other peroxisome proliferators (ciprofibrate and tibric acid) and also by oleoyl-CoA but by palmitoyl-CoA. Direct binding studies show that 35-40-kDa polypeptides bind oleoyl-CoA but not oleic or palmitic acid, or palmitoyl-CoA. 3. Polypeptides of 10-14 and 65-70 kDa also bind nafenopin-CoA. However, in contrast with 35-40-kDa polypeptides they also bind oleic and palmitic acid as well as their correspondent acyl-CoA thioesters. PMID- 8719906 TI - Influence of partial hepatectomy in dogs on trimethadione metabolism and microsomal monooxygenases. AB - 1. The recovery of trimethadione (TMO) metabolism and its association with liver weight and the activity of TMO N-demethylase have been reported in rat following partial (68%) hepatectomy. In the present study, we examined the effect of liver regeneration on hepatic P450 isozymes and TMO metabolism in dog. 2. The ratio of dimethadione (DMO), being the only TMO metabolite, to TMO at 2 h after i.v. injection of TMO (4 mg/kg) fell to 80% of that in the preoperative animals by 24 h after hepatectomy. The DMO/TMO ratio gradually recovered from days 7 to 14, and by day 21 after hepatectomy it had increased to about 25%. At 28 days post hepatectomy the ratio had returned to preoperative levels. 3. The activity of benzphetamine N-demethylase, TMO N-demethylase, p-nitro-anisole O-demethylase and aniline hydroxylase increased 3 days post-hepatectomy, exhibiting levels 4.77, 3.45, 1.51 and 1.91 times greater respectively than that of the preoperative liver in the same animal. Two weeks post-hepatectomy these activities had returned to normal. The activity of the 16 beta- and 2 beta-hydroxylation of testosterone was unchanged. However, the activity of 6 beta-hydroxylase decreased 7 days post-hepatectomy, while 16 alpha-hydroxylation had increased at 3 and 7 days post-hepatectomy compared with controls. 4. The changes in liver weight were nearly restored to preoperative levels 7 days post-hepatectomy. 5. Although the P450 content was unchanged from days 1 to 7 post-hepatectomy, it had decreased by 30% at day 14 and by 20% at day 28. The P4502B11 content 3, 7 and 14 days post hepatectomy had increased 8, 10 and 2 times respectively, while the P4503A12 content at 7 and 14 days decreased by 30 approximately 50% compared with that of the pre-operative liver. 6. The data presented above do not reveal any relationship between P4502B11 induction and liver regeneration. The reason for such a change is unknown, therefore further investigation needs to be carried out. PMID- 8719907 TI - Stereoselective reversible ketone formation from 10-hydroxylated nortriptyline metabolites in human liver. AB - 1. E- and Z-10-hydroxynortriptyline are major metabolites of amitriptyline and nortriptyline in man. Upon incubation with human liver microsomes or cytosol, these metabolites were oxidized to the corresponding ketones, E- and Z-10 oxonortriptyline. (+)-E- and (+)-Z-10-hydroxynortriptyline were distinctly preferred over the (-)-isomers as substrates. NADP+ supported the oxidation in cytosol, whereas in microsomes NAD+ was the best cofactor. 2. Incubation of E- and Z-10-oxonortriptyline with NADPH and cytosol resulted in the nearly exclusive formation of (+)-E- and (+)-Z-10-hydroxynortriptyline. Kinetic analysis revealed high-affinity reduction (K(m) 1-2 microM) of the two ketones and an additional low-affinity component with the E-isomer. 10-Oxonortriptyline reduction was also catalysed by rabbit, but not by rat or guinea pig liver cytosol. 3. With [4 3H]NADPH as cosubstrate, tritium was incorporated into E- and Z-10 hydroxynortriptyline preferentially from the pro-4R position. Redox cycling of (+)-E- and (+)-Z-10-hydroxynortriptyline in cytosol in the presence of NAD- and NADPH was indicated by 3H incorporation from [pro-4R-3H]NADPH. 4. Recombinant human carbonyl reductase catalysed low-affinity reduction of E-10 oxonortriptyline with preferential transfer of the pro-4S-3H of labelled NADPH. 5. Ketone reduction in cytosol was strongly inhibited by 9,10-phenanthrenequinone and dehydrolithocholic acid and moderately by other 3-oxo steroids and some anti inflammatory drugs. 6. The high-affinity reduction of E- and Z-10 oxonortriptyline and the oxidation of the alcohols in cytosol are probably mediated by a member of the aldo-keto reductase family of enzymes. PMID- 8719908 TI - Metabolism of sertindole: identification of the metabolites in the rat and dog, and species comparison of liver microsomal metabolism. AB - 1. The main exretion pathways of a novel antipsychotic drug, sertindole, in the rat and dog are faecal excretion via intestinal secretion and biliary excretion respectively. 2. Similar liver microsomal metabolic patterns were observed in the rat, monkey, and man, and Lu 30-131 (5-hydroxy-serindole) and Lu 30-148 (4 hydroxy-serindole) were the major metabolites, and Lu 25-073 (nor-sertindole) and Lu 28-092 (dehydro-sertindole) were minor ones. In the dog, however, Lu 31-096 (3'-fluoro-4'-hydroxy-sertindole) and Lu 30-148 (4-hydroxy-sertindole) were the major metabolites, and Lu 25-073 (nor-sertindole), Lu 28-092 (dehydro sertindole), and Lu 30-131 (5-hydroxy-sertindole) were minor ones. These findings suggest that the metabolism of sertindole in man resembles those in the rat and monkey and is different from that in the dog. 3. Rat in vitro and in vivo liver metabolites, dog liver microsomal metabolites, and dog biliary metabolites were isolated and identified by liquid chromatography/mass spectrometry and/or 1H-nmr. 4. Two metabolites, Lu 31-096 (3'-fluoro-4'-hydroxy-sertindole) and Lu 31-154 (3' fluoro-4'-hydroxy-dehydro-sertindole), were formed via the 'NIH shift' mechanism. 5. Sertindole is metabolized by hydroxylation at the 4- and 5-positions on the imidazolidinone ring, N-dealkylation, and an NIH shift at the fluorophenyl group. Further metabolism (dehydration, oxidation, hydroxylation, glucuronidation and sulphation) was also observed. 6. In the rat, oxidation at the imidazolidinone ring and N-dealkylation are the main metabolic reactions. On the other hand, in the dog, the NIH shift at the fluorophenyl group, followed by conjugation is the main metabolic pathway. PMID- 8719909 TI - Gliclazide hydroxylation by rat liver microsomes. AB - 1. The metabolism of gliclazide to hydroxygliclazide has been investigated in Sprague-Dawley rat liver microsomes. 2. The kinetics of hydroxygliclazide formation are consistent with Michaelis-Menten kinetics (mean (+/- SD, n = 3) apparent K(m) and Vmax = 256 +/- 27 microM and 1.85 +/- 0.10 nmol/ min/mg respectively). 3. Tolbutamide competitively inhibited hydroxygliclazide formation (Ki = 840 microM) and gliclazide competitively inhibited hydroxytolbutamide formation (Ki = 240 microM) with Ki similar to K(m). Therefore gliclazide and tolbutamide may be metabolized by the same enzyme in the rat. In nine livers the formation of hydroxygliclazide correlated with the formation of hydroxytolbutamide (rs = 0.82, p < 0.01). 4. Diclofenac (Ki = 64 microM), phenytoin (Ki = 38 microM), mephenytoin (Ki = 66 microM), glibenclamide (Ki = 14 microM) and glipizide (Ki = 189 microM) were fully competitive inhibitors of gliclazide hydroxylation. The rank order of Ki constants differed for gliclazide and tolbutamide suggesting that gliclazide and tolbutamide hydroxylases are not identical enzymes. 5. Quinine (Ki = 0.3 microM) and quinidine (Ki = 4.3 microM) were partially competitive inhibitors of hydroxygliclazide formation. Hydroxylation of gliclazide was related to the activity of CYP2D1 as assessed by dextrorphan production from dextromethorphan (rs = 0.83, p = 0.01). 6. In the rat gliclazide is metabolized to hydroxygliclazide by at least two cytochrome P450 isoforms, including tolbutamide hydroxylase and 2D1, which have similar affinities for gliclazide. PMID- 8719910 TI - Molecular orbital-generated QSARs in a homologous series of alkoxyresorufins and studies of their interactive docking with P450s. AB - 1. Molecular and electronic structural parameters have been determined, by molecular orbital (MO) calculations, for a homologous series of 8 alkoxyresorufins (methoxy- to octoxy-). 2. Quantitative structure-activity relationships (QSARs) between these structural parameters and the rates of metabolism of the alkoxyresorufins in hepatic microsomes from the 3 methylcholanthrene (MC)-, and phenobarbital (PB)-pretreated mouse, and the beta naphthoflavone (beta NF)-pretreated rat have been established. 3. The most significant single relationship is between beta NF-induction of cytochrome P4501 (CYP1A) and the total nucleophilic superdelocalizability (sigma SN) for the eight compounds in the series. 4. For double regressions, the electronic charge on the alkoxy oxygen, Q(O), or alpha-carbon Q(C), is important when combined with the hydrophobic substituent constant (pi). 5. These findings indicate that the rates of metabolism of these alkoxyresorufins are dependent upon their ability to cross cellular membranes, to fit the relevant CYP1A binding site, and on their ability to accept electrons from a donor nucleophilic species. 6. A different set of parameters correlated with CYP2B activity, namely, parameters of overall shape, which indicates that the way in which the alkoxyresorufins fit the CYP2B site, determines their differences in specificity. 7. Computer graphic interactive docking studies of the alkoxyresorufins with their affinity-specific cytochromes P450, namely, methoxy- with CYP1A2; ethoxy- with CYP1A1; pentoxy- with CYP2B1; and benzyloxy- with CYP3A, have also been undertaken to show the specific interactions of the alkoxyresorufins with the binding sites of the individual P450s. PMID- 8719911 TI - Pharmacokinetics of the 5-hydroxytryptamine1A agonist 8-hydroxy-2-(N,N-di-n propylamino)tetralin (8-OHDPAT) in the rat after intravenous and oral administration. AB - 1. Plasma levels of 3H and unchanged drug were measured in the non-anaesthetized male rat after intravenous (i.v.) or oral administration of (+/-)-(R,S)-[propyl 3H]-8-OHD-PAT, at three dose levels per route of administration. The excretion of conjugated metabolites in bile was also studied following i.v. administration. 2. For unchanged 8-OHDPAT following i.v. administration, terminal t1/2 was 1.56 +/- 0.01 h (mean +/- SD, n > or = 4), kelim 0.45 +/- 0.01 h-1, volume of distribution 0.14 +/- 0.02 litres and clearance 1.10 +/- 0.17 mlmin-1. After oral administration, terminal t1/2, kelim, apparent volume of distribution and clearance were essentially the same when bioavailability was taken into account. Neither dose size nor route of administration had any significant effect on either terminal t1/2 or kelim. Comparison of AUCs following i.v. and oral administration yielded a mean for absolute oral bioavailability of 2.60 +/- 0.24%. 3. Comparison of AUCB for total plasma 3H showed that the extent of absorption was 80.1%, indicating that the low oral bioavailability of 8-OHDPAT is due to first-pass metabolism, rather than poor absorption from the GI tract. 4. Following i.v. administration, irrespective of dose, some 10% of the 3H dose was excreted in the bile in 6 h, 8.5% as 8-OHDPAT-glucuronide and 1.5% as the glucuronide of the N-despropylated metabolite, 8-OHDPAT. The majority of the biliary excretion occurred within 3 h of dosing. PMID- 8719912 TI - Bosentan, a new endothelin receptor antagonist: prediction of the systemic plasma clearance in man from combined in vivo and in vitro data. AB - 1. Accurately predicting the kinetics in man greatly improves the design of the phase I clinical studies. This was particularly crucial in the case of bosentan, a new endothelin receptor antagonist, as very large interspecies differences in systemic clearance were observed in the animal species investigated, namely from 1.5ml/min/kg in the dog up to 70 ml/min/kg for the rabbit. 2. Bosentan was shown to be metabolized by the hepatic cytochrome P450, therefore the rate of metabolism was investigated in vitro in liver microsomes and hepatocytes, across the species which had been tested in vivo. The same rank-order of metabolism was found for the laboratory animals both in vitro and in vivo, and hepatocytes appeared to be more representative of the in vivo situation than liver microsomes. The in vitro clearance in human hepatocytes was very close to that observed in dog hepatocytes. 3. A plasma clearance for bosentan in man of 1-2 ml/min/kg was predicted by combining the in vivo and in vitro data from a few animal species with the in vitro data in man. This expectation was subsequently found to agree reasonably well with the plasma clearance observed in healthy volunteers: ca 2 ml/min/kg. Integrating this prediction into the design of the first clinical protocols substantially improved the quality of the human pharmacokinetic data obtained. PMID- 8719913 TI - Fate of dipropyl sulphone in rat. AB - 1. Dipropyl [35S]-sulphone was administered by gavage (4.24 mmol/4 ml/kg body weight) to the adult male Wistar rat following an overnight fast. 2. Urine was the major route of excretion (83%) with more radioactivity appearing during the second day (47%) than the first (28%). Only small amounts were found in the faeces (10%). Biliary excretion played an important role with substantial amounts of the dose (33%) passing through the bile duct during 0-48 h. A near total recovery was achieved suggesting that only small amounts (2%) may have been lost as volatile components. 3. Metabolism was limited, the majority (> or = 98%) of the sulphone being recovered unchanged. Oxidation of the sulphur with the formation of inorganic sulphate was the only pathway observed. PMID- 8719914 TI - Role of cytochrome P450 in hydrazine toxicity in isolated hepatocytes in vitro. AB - 1. Hepatocytes, isolated from the control, diethyldithiocarbamate (DEDC), acetone, isoniazed and hydrazine pretreated rat, were incubated with hydrazine (8 20 mM) for 3 h. Hydrazine caused a dose-dependent loss of viability, leakage of LDH, depletion of GSH and ATP and an inhibition of the incorporation of 3H leucine into protein. 2. Pretreatment with DEDC increased, whereas hydrazine and acetone pretreatments decreased the cytoxicity and biochemical effects of hydrazine. Pretreatment with isoniazid slightly increased hydrazine cytotoxicity. Acetone pretreatment reduced the inhibition of protein synthesis caused by hydrazine compared to the control. 3. 4-Nitrophenol hydroxylase activity (P4502E1) correlated with viability, LDH leakage, ATP and GSH depletion in cells from the control, DEDC, acetone and hydrazine pretreated rats. 4. The activities of PROD (P4502B1) and EROD (P4501A1/1A2) also correlated with the above parameters for all treatments. The results suggest that three isoenzymes may be involved in the detoxication of hydrazine. Protein synthesis inhibition did not correlate with the activities of any of the enzymes measured. PMID- 8719915 TI - Central neural organization and control of sympathetic nervous system in mammals. AB - The past decade has witnessed rapid progress in defining neural circuits and mechanisms in the brain, responsible for regulation of the sympathetic nerve activity and cardiovascular functions. Several groups of cardiovascular neurons in the brainstem form the fundamental neural circuits, through which reflexly and centrally initiated sympathetic responses are processed. Their interplay determines the levels of sympathetic nerve activity and vascular tone. Substantial evidence indicates that a small population of reticulospinal vasomotor neurons in the rostroventrolateral reticular nucleus of the medulla oblongata play critical and integrative roles by: 1) providing, largely by their intrinsic pacemaker activity, tonic sympathoexcitation, thus maintaining normal blood pressure and organ blood flows, 2) mediating a variety of circulatory reflexes and centrally initiated sympathetic responses thereby helping to match organ blood flow to metabolic demands, and 3) acting as intrinsic oxygen detectors which orchestrate appropriate autonomic response programs to protect the integrity of brain in response to acute hypoxia-ischemia. Elaboration of the neural mechanisms and cellular and molecular properties of these vasomotor neurons related to dynamic regulation of the cardiovascular system in normal and disease states will be of relevance to a full appreciation of their role in adaptation of the organism to its internal and external environments and to the development of strategies to fight against neurogenic cardiovascular diseases and to restore normal functions. PMID- 8719916 TI - Axonal sprouting and synaptogenesis in temporal lobe epilepsy: possible pathogenetic and therapeutic roles of neurite growth inhibitory factors. AB - Axonal sprouting and synaptic reorganization within the temporal lobe following neuronal injury have been implicated in the pathogenesis of temporal lobe epilepsy (TLE). The molecular species responsible for these structural changes have yet to be fully defined. The recent characterization of molecules whose normal function within the nervous system is to inhibit neurite outgrowth and synaptogenesis prompts the suggestion that a diminution or loss of such molecules might be of relevance to the pathogenesis of TLE. If so, the possibility of developing a novel therapeutic approach to TLE, distinct from currently available symptomatic therapies, to arrest the pathogenetic processes is also raised. PMID- 8719917 TI - Tics and fits. The current status of Gilles de la Tourette syndrome and its relationship with epilepsy. AB - The Gilles de la Tourette syndrome (GTS) is a neurobehavioural disorder characterized by multiple motor and vocal tics. Accessory symptoms include coprophenomena, echophenomena and obsessive-compulsive behaviours. There is a significant genetic contribution to their aetiology. Simple tics may be mistaken for myoclonic epilepsy and complex tics for complex partial seizures. Like epilepsy, GTS is a brain disorder that leads to general disturbances in behaviour not just the cardinal symptoms (i.e. tics or seizures). PMID- 8719918 TI - Mechanisms of action of antiepileptic drugs. AB - Depending on their mechanism of action, anticonvulsant drugs in clinical use may be divided into three groups: those drugs which facilitate gamma-aminobutryic acid (GABA)ergic neurotransmission; those which block neuronal ion channels; and those whose mechanism of action is unresolved. The compounds acting on GABAergic systems may be further subdivided into those which modulate transmission through chloride channels, e.g. the barbiturates and the benzodiazepines; those compounds, in particular vigabatrin, which reduce the degradation of GABA by blocking GABA transaminase; and those which inhibit the re-uptake of GABA into the presynaptic terminal. The other group of compounds whose mechanism of action is known are those which block neuronal ion channels. Blockage of voltage operated sodium channels by lamotrigine, phenytoin or carbamazepine leads to decreased electrical activity and, probably, a subsequent reduction in glutamate release. Conversely, ethosuximide, blocks voltage-operated calcium channels, especially those which mediate calcium currents in thalamic neurones. Of those drugs in which the mechanism of action is unknown, sodium valproate is the prime example. An antagonistic action at the N-methyl-D-aspartate (NMDA) subtype of glutamate receptor might also be a possibility, which could be the case with some of the newer compounds currently undergoing evaluation. PMID- 8719919 TI - Mortality related to convulsive disorders in a developing country in Asia: trends over 20 years. AB - The cause of death as recorded in 37125 death certificates (DC) issued in the Kandy District over 20 years at five-year intervals beginning 1967 was analysed to determine the trends in mortality caused by convulsive disorders in the community. Convulsions accounted for 881 (23.7/1000) deaths, the highest number being in infancy (35.8%). A slight male preponderance of 51.5% was observed. Most of the deaths occurred in the periphery (51.6%) and in the tea estates (36.3%) as opposed to town area (12.0%). "Febrile convulsions' was the most common diagnosis in 396 (44.9%) deaths. "Convulsions' which included neonatal and infantile convulsions accounted for 186 (21.1%) deaths. Other causes included chest complications (60, 6.8%), drowning (28, 3.2%), asphyxia (20, 2.3%), status epilepticus (19, 2.2%), burns (7, 0.8%), and poisoning (2, 0.2%). The proportion of deaths due to convulsive disorders showed a decrease from 37.28/1000 in 1967 to 9.55/1000 in 1987, which was most evident in the periphery, from 60.49/1000 to 13.19/1000. This parallelled a drop in the proportion of deaths attributed to "febrile convulsions' and "convulsions'. The study shows the need to educate the public about first-aid and the subsequent management of convulsions, especially in childhood. Health personnel in developing countries should consider it mandatory to make a specific diagnosis in children who present as febrile convulsions. Appropriate antiepileptic medication and compliance can prevent death due to status epilepticus and injuries associated with seizures. PMID- 8719920 TI - Epilepsy in a children's hospital: an out-patient survey. AB - Epilepsy, a common childhood condition, is largely managed by general paediatricians. To assess whether recent management guidelines are being met, we undertook a prospective questionnaire-based survey of children with epilepsy, attending the general paediatric out-patient clinic, at the Royal Liverpool Children's Hospital, Alder Hey. Ninety-one children were identified over seven months-giving a prevalence of epilepsy in this out-patient population of 13.7/1000. Fifty-four per cent were diagnosed as having tonic-clonic, 16% partial and 5% typical absence seizures. Forty-two per cent received carbamazepine and 33% sodium valproate, as monotherapy, and 7% were treated with multiple antiepileptic drugs (AEDs). Correlation between AED and seizure type was often inappropriate as was the routine measurement of blood level of AEDs. Correlation between seizure frequency and clinic visits was generally acceptable with those children experiencing frequent seizures attending most often. Specialist paediatric neurologist advice was sought in only 14% of children; 19 of the 28 children with the most poorly controlled seizures were not referred for a specialist opinion. This study has demonstrated a poor understanding of seizure/syndrome classification; the frequent inappropriate use of AEDs and their blood level measurement and inadequate referral for specialist advice. Recognized guidelines are not being followed, nor are standards being met by many general paediatricians. There is a clear indication for more extensive education and the need to provide clear recommendations for the management of epilepsy in children. PMID- 8719921 TI - The management of epilepsy in children: the role of the clinical nurse specialist. AB - Epilepsy in children is frequently accompanied by learning, behavioural and psychological difficulties. These difficulties may be compounded by the ignorance and stigma which exists within the community health care and education services. The management of epilepsy in children therefore extends far beyond the use of antiepileptic drugs and must address these additional problems. The establishment of a nurse specialist service in paediatric epilepsy within our hospital has provided a more satisfactory and comprehensive management of these children and, in addition, has facilitated a close liaison with schools, community health personnel and support groups; as a result this has dispelled much of the local ignorance, misunderstanding and stigma surrounding epilepsy. PMID- 8719922 TI - Monotherapy antiepileptic drug trials in patients undergoing presurgical assessment: methodological problems and possibilities. PMID- 8719923 TI - Natural atmospherics and occurrence of seizures in six adolescents with epilepsy: a cross correlation study. AB - As already seen in a former study of 315 epileptic seizures in adults, subsequent investigation of 3333 epileptic seizures in six adolescents revealed a significant increase of the seizure frequency during days with a higher mean frequency of 28 kHz atmospherics, and a decrease during days with a (Baumer apparatus) distinctly higher amount of 10 kHz when compared with the daily mean frequency within the whole period. However, one patient showed an opposite behaviour regarding the correlations of 28 and 10 kHz atmospherics and the mean numbers of seizures. PMID- 8719924 TI - Non-epileptic attack disorders in children and adolescents: a single case study. AB - Previous research into non-epileptic attack disorder (NEAD) has focused on the individual. We report a teenager with non-epileptic seizures which highlights the importance of the family in the development and maintenance of such attacks. The authors propose that in children and adolescents the focus of therapeutic intervention should be the individual within the context of the family. PMID- 8719925 TI - Transient global amnesia of epileptic origin accompanied by fever. AB - The case of a previously healthy 69-year-old female patient is described who presented, in a period of 6 months, 3 long-lasting (from 2 hour- to 10 hour duration) episodes of transient global amnesia accompanied by a temperature rise. During one of these episodes an EEG was obtained, showing a diffuse alteration, focal slowing, and bitemporal asynchronous paroxysmal activity giving rise to electrical ictal discharges. Interictal EEGs were normal. Cerebral computed tomography was normal. Carbamazepine was given with complete control of the attacks. These episodes may be interpreted as complex partial status with unusual semeiology. PMID- 8719926 TI - The reproducibility and comparability of tests of the peripheral chemoreflex: comparing the transient hypoxic ventilatory drive test and the single-breath carbon dioxide response test in healthy subjects. AB - Both the transient hypoxic ventilatory drive test and the single-breath carbon dioxide (CO2) response test have been used to assess peripheral chemoreflex sensitivity. We tested their comparability in 14 healthy adults (10 men, aged 31 73 years, mean 55.4 years). The within-subject reproducibility of both tests was also assessed (n = 7 for each). The mean transient hypoxic ventilatory response was 0.287 +/- 0.0591 min-1 (%Sao2)-1 (mean +/- SEM, range 0.018- 0.718) and single-breath CO2 response was 0.276 +/- 0.0411 min-1T-1 (range 0.081-0.501). Both tests were reproducible with a mean coefficient of variation of 20.1% and 17.7%, respectively. There was, however, no significant correlation between the results of the transient hypoxic and single-breath CO2 tests when data were compared by linear regression analysis (r = 0.23, P = 0.43), suggesting that separate pathways of the peripheral chemoreflex existed for hypoxia and hypercapnia, respectively, and that these tests were specific for each. The authors conclude that these tests are reproducible but need to be used in combination for an adequate assessment of the peripheral chemoreflex. PMID- 8719927 TI - Relation of endothelins to volume regulating neurohumoral systems in patients with cirrhosis of the liver. AB - The aim of the present study was to investigate the relationship between ET plasma concentrations and other hormonal systems in acute volume regulation of patients with cirrhosis. Ten healthy controls and 10 cirrhotic patients, five without and five with ascites were studied after 1 h in a sitting posture and subsequently subjected to 1 h head-out water immersion. Blood was collected for determinations of ET-1, ET-3, ANF, aldosterone, renin activity and noradrenaline. In addition, in 10 patients with compensated cirrhosis the effect of loop diuretics on ET-3, aldosterone and renin was studied. ETs in cirrhosis were significantly (P < 0.01) higher than in controls both before (ET-1, 19.6 +/- 1.3 pgmL-1 vs. 11.8 +/- 0.4 pgmL-1; ET-3, 18.5 +/- 1.4 pgmL-1 vs. 9.5 +/- 0.5 pgmL-1) and after water immersion (ET-1, 18.6 +/- 1.2 pgmL-1 vs. 12.4 +/- 0.3 pgmL-1; ET 3, 18.7 +/- 1.7 pgmL-1 vs. 10.0 +/- 0.5 pgmL-1). In cirrhotic patients, basal and immersion concentrations of ET-1 were significantly correlated to noradrenaline plasma concentrations (r = 0.79, P < 0.05). ET-3 plasma concentrations in cirrhosis were correlated to renin activity (r = 0.65, P < 0.05). Furthermore, ET 3 in cirrhosis was inversely correlated to systolic and mean arterial blood pressure (r = -0.55, P < 0.01 and r = -0.50, P < 0.05; respectively). To investigate the effect of hypovolaemia in compensated cirrhosis, 10 patients without ascites were studied before and after treatment with loop diuretics. In compensated cirrhosis ET-3 was significantly increased 6h after oral diuretic treatment (17.9 +/- 1.0 pgmL-1 vs. 15.5 +/- 0.4 pgmL-1, P < 0.001). The presented data demonstrate relations of endothelins, particularly of ET-3 to neurohumoral systems in patients with cirrhosis of the liver. PMID- 8719928 TI - Circulating and tissue guanylin immunoreactivity in intestinal secretory diarrhoea. AB - Guanylin is a recently discovered peptide hormone that activates intestinal guanylate cyclase (GC-C) and thereby stimulates intestinal chloride secretion. Immunohistochemistry showed its presence in enterochromaffin (EC) cells of the gut. In vitro studies suggested that guanylin plays an important role in the endogenous modulation of intestinal salt and water secretion. In the present study the concentration of circulating immunoreactive (IR)-guanylin in plasma of patients with intestinal diarrhoea due to chronic bowel inflammation and patients with carcinoid tumours were measured with a specific radioimmunoassay. In 22 patients with Crohn's disease and eight patients with ulcerative colitis, plasma concentrations of IR-guanylin were 44 +/- 3 and 42 +/- 4 fmol mL-1, respectively. Levels were not different from that in 44 healthy volunteers suggesting that the circulating hormone is not involved in diarrhoea of these patients. In 17 patients with symptomatic carcinoid tumors the median concentration of circulating IR-guanylin was significantly enhanced (94 +/- 16 fmol mL-1, range 37 312 fmol mL-1). Immunohistochemistry revealed the presence of immunoreactive guanylin in carcinoid tissues, suggesting that these tumours co-release guanylin along with their usual resident hormone, serotonin. Enhanced local secretion of guanylin may play a causal role in diarrhoea of these patients and its elevation in plasma may be of diagnostic value in this type of endocrine tumours. PMID- 8719929 TI - Recurrent condylomata acuminata: how routine immediate and delayed hypersensitivity parameters might provide a clue to their immunopathogenesis. AB - In 30 male patients suffering from recurrent condylomata acuminata, immediate hypersensitivity parameters (total IgE, PTT and prick tests) and delayed hypersensitivity against seven recall antigens (multi test) were studied. Thirty healthy male volunteers, matched in age, were the controls. Significantly higher immediate hypersensitivity activity was shown in the patient group. Qualitative evaluation of delayed type hypersensitivity showed that controls had a positive test 16 times more often than patients. A rather homogeneous suppression of delayed type hypersensitivity was found in the patient group mainly as regards the presumably most common antigens vs. the control group. This suppression was proved to be related to disease duration. The hypothesis of a CD4+ Th-2 lymphocyte predominance in recurrent condylomata, owed to longstanding or repetitive antigenic stimulation seems to adequately explain the findings of the present study. PMID- 8719930 TI - Lichen planus and hepatitis C virus in the northern Kyushu region of Japan. AB - Oral lichen planus (OLP) is a common oral disorder that manifests a mucosal reaction to a variety of aetiological factors, including liver disorder. This study investigated the relationship between OLP and hepatitis C virus (HCV) infection by studying the prevalence of hepatitis B and C virus infection or liver disease in 45 patients with OLP in the Northern Kyushu region of Japan where the prevalence of HCV infection is the highest in the country. Serum hepatitis B virus surface antigen (HBsAg) was positive in only four patients. Serum anti-HCV or serum HCV RNA was positive in 28 (62%) and 27 (60%) of 45 OLP patients, respectively. The majority (35 of 45, 78%) of OLP patients suffered from liver disease, including chronic hepatitis C (22/45, 49%), HCV-related liver cirrhosis (two), and HCV-related hepatocellular carcinoma (two). These results suggest that HCV is a major cause of OLP. PMID- 8719931 TI - Omega-3 fatty acids suppress the enhanced production of 5-lipoxygenase products from polymorph neutrophil granulocytes in cystic fibrosis. AB - Pulmonary damage in cystic br osis (CF) is associated with chronic inflammation mediated in part by proinflammatory 5-lipoxygenase products (5-LOP, leukotrienes and 5-hydroxyeicosatetraenoic acid) from polymorph neutrophil granulocytes (PMN). The authors studied 5-LOP formation of PMN from CF patients and in vitro effects of added eicosapentaenoic acid (EPA) and fish oil. Circulating PMN were isolated from 10 CF patients without acute infections and 10 control persons of the same age (4-20 years). Total 5-LOP liberation from PMN of CF patients was significantly increased over controls after incubation with the calcium ionophore A23 (1 mumol L-1) without arachidonic acid (AA) (380 +/- 24 vs. 294 +/- 28 pmol mL-1) and with 10 mumol L-1 AA (1303 +/- 104 vs. 1015 +/- 104 pmol mL-1), and there were nonsignificant trends to high values after incubation with 5 mumol L-1 platelet activating factor (PAF, 134% of controls) and 1 mumol L-1 formyl methionylleucyl-phenylalanine (FMLP, 125%). The addition of 100 micrograms mL-1 fish oil to PMN of CF patients challenged with A23 completely suppressed synthesis of proinflammatory 5-LOP of the 4-series, while inactive 5-LOP metabolites of the 5-series were produced. Added EPA (10 mumol L-1) also suppressed 4-series 5-LOP and significantly reduced leukotriene B4 concentration by 48% from 39.9 +/- 3.2 to 20.6 +/- 11.4 pmol L-1, again with a concomittant increase of inactive 5-series metabolites. The authors conclude that the turnover of endogenous and exogenous AA is enhanced in CF, possibly due to stimulated phospholipase A2 activity. The relatively small effect of the receptor dependent stimuli PAF and FMLP may be caused by a down-regulation of PMN receptors in CF. Supplementation of long-chain omega-3-fatty acids may be beneficial for reducing excessive inflammation in CF patients and should be further evaluated. PMID- 8719932 TI - A comparison of the effects of insulin-like growth factor-I, insulin and combined infusions of insulin and insulin-like growth factor-I on glucose metabolism in dogs. AB - The effect of infusions of recombinant insulin-like growth factor-I (IGF-I) (34, 103 or 688 pmol kg-1 min-1), insulin (3.4, 10.3 or 68.8 pmol kg-1 min-1) or combined infusions (34 pmol IGF-I + 3.4 pmol kg-1 min-1 insulin or 103 pmol IGF-I + 3.4 pmol kg-1 min-1 insulin) on glucose metabolism was investigated in dogs using a [3-3H]-glucose infusion and euglycaemic clamp. All insulin doses decreased glucose production rate (Ra) in a dose-dependent manner (P < 0.05). All IGF-I doses decreased glucose Ra (P < 0.05) but this decrease was not dose dependent. The decrease in glucose Ra with the combined infusion of 34 pmol kg-1 min-1 IGF-I + 3.4 pmol kg-1 min-1 insulin was greater than 34 pmol kg-1 min-1 IGF I (P < 0.05) but not different from 3-4 pmol kg-1 min-1 insulin. All insulin and IGF-I doses increased glucose utilization rate (Rd) in a dose-dependent manner (P < 0.01). The increase in glucose utilization was greater following both combined infusions than with either component infused alone (P < 0.05). Although at the doses selected, insulin and IGF-I had similar effects on glucose utilization with additive effects when the two peptides were combined, IGF-I was less effective than insulin in suppressing glucose production. PMID- 8719933 TI - Interactions between platelets and neutrophils in essential thrombocythaemia. Effects on neutrophil chemiluminescence and superoxide anion generation. AB - Essential thrombocythaemia (ET) is frequently associated with neutrophil and platelet dysfunction, and with increased incidence of vascular complications (thrombosis, haemorrhage). Several interactions between platelets and neutrophils have been reported, and the reciprocal actions between these cells may have an important role both in thromboregulation and in diseases such as those caused by uncontrolled neutrophil activation. In the current paper the authors studied 15 patients affected by ET and 10 normal subjects as controls. Circulating neutrophils and platelets were purified and were recombined in constant ratios (50:1, 100:1 and 200:1) and the individual platelet to neutrophil ratio. Superoxide anion (O2-) generation and luminol-enhanced chemiluminescence (CL) were studied after neutrophil stimulation with fMLP. In normal subjects both O2- generation and CL were inhibited by autologous platelets in a dose-dependent manner. In ET patients, on the contrary, platelet-dependent inhibition of O2- generation did not occur, while a dose-dependent inhibition of CL was observed. Two groups of ET patients were found: patients with neutrophil O2- generation and CL within the normal range, and patients with significantly reduced neutrophil respiratory burst. However, no differences were found between these two groups of patients in terms of platelet effects towards fMLP-stimulated neutrophils. Therefore, platelets from ET patients were not able to exert the homeostatic control towards neutrophil O2- generation shown by platelets from normal subjects, and this phenomenon may have a role in the clinical setting. In fact, O2- has been shown to be a very strong direct platelet activator, is able to inactivate nitric oxide (which is a powerful inhibitor of platelet aggregation and adhesion to endothelium), and is directly involved in neutrophil-mediated tissue damage. PMID- 8719934 TI - Progression of coronary artery disease in non-dilated sites in the months following balloon angioplasty: time-dependent relation with restenosis. AB - There is scant information on the progression of coronary artery disease in non dilated sites in the months following percutaneous transluminal coronary angioplasty (PTCA) or on its relationship with restenosis. To assess the incidence of this progression and its relationship with restenosis at various times after PTCA, the authors selected 371 consecutive patients who had undergone a first successful PTCA for angina on native coronaries followed by a repeat angiographic study. The angiograms were analysed by a computer-assisted method; progression was defined as a 20% decrease in diameter and restenosis as a 30% decrease in diameter or a return to > 50% stenosis. The relationship between progression and restenosis was analysed in the whole population and then, using the Mantel-Haenszel chi-square test, in two subgroups: patients with a stable clinical state, who were restudied routinely and those whose worsened state had prompted repeat angiography. The relationship was assessed at different times between angioplasty and the repeat angiography. Progression was observed in 80 patients (22%) and restenosis in 155 patients (42%). There was a highly significant relationship between progression and restenosis in the total population (chi 2 = 26.4, odds ratio = 3.9 and P < 0.0003) and in the group of patients that were routinely restudied (chi 2 = 31.6, odds ratio = 5.3 and P < 0.0001), but not in the group of patients in whom restudy was performed because of clinical worsening (chi 2 = 0.13, odds ratio = 1.5 and P = NS). With respect to the length of follow-up, in the total population the relationship was significant only at 6 and 7 months (P < 0.0001), and in the group receiving a routine restudy only at 4-5 and 6-7 months (P < 0.001). Progression in non dilated sites appeared to be strongly and transiently linked with restenosis, suggesting that PTCA may enhance both restenosis and progression over a short period. PMID- 8719935 TI - Effect of danazol-induced chronic hyperglucagonaemia on glucose tolerance and turnover. AB - It has been shown that danazol (14-ethinyltestosterone) induces hyperglucagonaemia. To investigate the effect of chronic glucagon excess on carbohydrate metabolism, we studied six patients before and after treatment with danazol for immunothrombopenia. Glucose tolerance and insulin, C-peptide and glucagon secretion during an oral glucose tolerance test (oGTT) as well as peripheral and hepatic insulin sensitivity were determined by means of euglycaemic clamp technique (40 mU m-2 min-1) before and after 3 months of danazol therapy. Overall glucose turnover (Rd) was assessed radioisotopically. (1) Plasma glucagon levels rose significantly from 88 +/- 16 pg mL-1 before to 683 +/- 148 pg mL-1 after therapy (P < 0.01). (2) Glucose levels during an oGTT were not significantly different before and after therapy. Glucose-stimulated insulin secretion at 60 and 120 min and the area under the curve (AUC) for insulin during the oGTT, were significantly increased after danazol treatment compared with pre-treatment values (P < 0.05), whereas glucagon secretion showed a similar decrease at both time points of investigation (NS). (3) Rd during steady state showed a significant decrease during the entire period of euglycaemic clamp following therapy (after 240 min, 3.8 +/- 0.6 vs. 5.3 +/- 0.7 mg kg-1 min-1, P < 0.05). The decline in glucagon during the clamp was similar during steady state before and after therapy. (4) Basal hepatic glucose output did not differ significantly before and after therapy (1.74 +/- 0.41 vs. 1.45 +/- 0.22 mg kg-1, NS), whereas hepatic glucose output during the clamp was significantly less suppressed after danazol therapy. The authors conclude that chronic glucagon excess leads to a decrease in peripheral and hepatic insulin action which is accompanied by an increase in insulin secretion. PMID- 8719936 TI - An aldose reductase inhibitor, TAT, prevents electroretinographic abnormalities and ADP-induced hyperaggregability in streptozotocin-induced diabetic rats. AB - Rats with streptozotocin-induced diabetes were oral given TAT, a potent aldose reductase inhibitor, at a dose of 10 mg kg-1 day-1 or 40 mg kg-1 day-1 for 30 days. Prolongation of the peak latency of oscillatory potentials in the b-wave of the electroretinogram (ERG), which is associated with retinal Muller cell dysfunction, was significantly improved by treatment with TAT as compared with untreated diabetic rats [sigma(O1 + O2 + O3) was 106.8 +/- 1.8 ms in normal controls (NC), 118.2 +/- 1.1 ms in diabetic controls (DC) (P < 0.001 vs. NC), 110.8 +/- 1.5 ms with 10 mg kg-1 TAT (P < 0.001 vs. DC) and 111.4 +/- 1.6 ms with 40 mg kg-1 TAT (P < 0.01 vs. DC)]. The improvement in ERG abnormalities in diabetic rats was accompanied by partial reduction of elevated sorbitol levels in the retina and erythrocytes, and by correction of platelet hyperaggregability. The authors' findings suggest that a better understanding of the mechanism by which TAT acts may provide new insights into the pathogenesis of hyperglycaemic retinal dysfunction and contribute to establishing effective therapy for diabetic retinopathy. PMID- 8719937 TI - Pre-operative evaluation of risk factors for complications in patients with primary hyperparathyroidism. AB - Impairments in cardiovascular, respiratory and kidney function are considered as risk factors for complications following surgery. As the indication for surgery in asymptomatic primary hyperparathyroidism (HPT) is controversial, 123 patients undergoing surgery for HPT and 104 control subjects scheduled for common surgical procedures were evaluated pre-operatively. Compared with the age- and sex-matched control group, serum calcium (P < 0.001), creatinine (P < 0.01) and glucose (P < 0.02) were all increased in the HPT group, while peak expiratory flow (PEF) was decreased (P < 0.04). Furthermore, the patients with HPT, compared with controls, were more often receiving antihypertensive medication (P < 0.005) and were more likely to have a history of congestive heart disease (P < 0.01), thromboembolic diseases (P = 0.05), stroke (P = 0.06) or diabetes mellitus (P < 0.02). Increased frequencies of ST-segment depression (P < 0.001) and T-wave abnormalities (P = 0.05) at electrocardiography together with an increased prevalence of heart enlargement visible at chest radiography (P < 0.01) were also seen in the HPT group when compared with the controls. All HPT patients and controls survived, but one HPT patient suffered a myocardial infarction in the post-operative period. In conclusion, the present study showed the pre-operative risk factor profile to be altered in HPT subjects with impairments in both cardiovascular and respiratory functions as well as in kidney function and glucose control. These findings should be kept in mind when the indications for surgery in asymptomatic patients with HPT are discussed. PMID- 8719939 TI - Is responsiveness to lovastatin in familial hypercholesterolaemia heterozygotes influenced by the specific mutation in the low-density lipoprotein receptor gene? AB - Lovastatin is one of the most commonly used lipid-lowering drugs in familial hypercholesterolaemia (FH) heterozygotes. In order to study whether the response to lovastatin is influenced by the underlying mutation in the low-density lipoprotein (LDL) receptor gene, the authors compared the response in 24 heterozygotes in whom the mutation has been classified and in 34 heterozygotes in whom the mutation has not been classified. Those possessing a classified mutation had significantly higher pre-trial values of LDL-cholesterol than those possessing an unclassified mutation. However, no difference was found in the response to lovastatin. Nor were there any differences in the response between subjects possessing one of the three different classified mutations. Furthermore, irrespective of whether or not the mutation had been classified, no difference in the response was found between subjects in the upper and lower quartile with respect to pre-trial values of LDL-cholesterol. The authors conclude that the response to lovastatin is independent of both the specific mutation in the LDL receptor gene and the actual cholesterol level in FH heterozygotes. PMID- 8719938 TI - Transforming growth factor beta 1 influences lysyl hydroxylation of collagen I and reduces steady-state levels of lysyl hydroxylase mRNA in human osteoblast like cells. AB - Transforming growth factor beta 1 (TGF-beta 1) is an osteotropic growth factor that is found in substantial concentration in bone. The authors studied the influence of TGF-beta 1 on the modification of lysine residues of collagen I. The degree of lysyl hydroxylation and lysyl glycosylation of newly synthesized collagen as well as steady-state levels of mRNA for both lysyl hydroxylase and collagens I and III were determined in human osteoblast-like cells in vitro. In normal human osteoblasts lysyl hydroxylation was decreased by TGF-beta 1 particularly in the collagen alpha 2-chain. This effect was paralleled by an increase in lysyl residues, whereas glycosylation was not affected. The mRNA for lysyl hydroxylase was reduced by one-third under the influence of TGF-beta 1. Additionally, the mRNAs for both procollagen I alpha-chains were stimulated by TGF-beta 1, whereas pro alpha 1 (III)-mRNA showed a decrease. Changes in the local regulatory activity of TGF-beta 1 may play a role in matrix maturation such as collagen type production and lysyl hydroxylation, the latter being altered in various pathological conditions, e.g. in generalized osteopenia. PMID- 8719940 TI - Vascular reactivity to noradrenaline and neuropeptide Y in the streptozotocin induced diabetic rat. AB - The study aimed to assess vascular reactivity to noradrenaline with and without neuropeptide Y in diabetic rats, and to determine whether any abnormality could be attributed to insulin deficiency or to hyperglycaemia per se. The authors compared non-diabetic rats (n = 9) and rats with streptozotocin-induced diabetes that were either untreated (n = 10), or treated with insulin (n = 9) or food restriction (n = 8) to restore near-normoglycaemia. After 4 weeks of diabetes, contractile responses to noradrenaline (0.24-48 mumol L-1), without and with neuropeptide Y (0.1 mumol L-1), were assessed using an isometric myograph in two mesenteric arteries from each rat. Vessels from untreated diabetic rats were significantly more reactive to noradrenaline than the control vessels when tested without (P < 0.0001) but not with (P = NS) neuropeptide Y. Diabetic rats rendered nearly normoglycaemic through food restriction showed dose-response curves that were very similar to the untreated diabetic group (P = NS). By contrast, insulin treated diabetic vessels showed reduced sensitivity to noradrenaline, with and without neuropeptide Y, compared with both the diet-restricted and untreated vessels (both P < 0.0001). The authors conclude that vascular sensitivity to noradrenaline, without or with neuropeptide Y, is reduced over a wide dose range in vessels taken from rats treated in vivo with insulin; furthermore, vessels taken from diabetic rats not treated with insulin (hypoinsulinaemic) tended to be more reactive than either control vessels or those taken from the insulin-treated rats. The latter group of rats were probably hyperinsulinaemic for much of the time; the results may therefore support the hypothesis that insulin acts as a vasodilator. PMID- 8719941 TI - A mathematical analysis of a two-compartment model of urea kinetics. AB - A model of urea distribution comprising well-mixed intracellular and extracellular compartments, with diffusive transfer of urea between compartments, is used to study blood urea variation during haemodialysis. Assuming that a typical adult patient (weight 70 kg and urea generation rate 5 mg min-1) is dialysed using a high-efficiency dialyser (urea clearance 0.2- 0.251 min-1) at an ultrafiltration rate of 10 ml min-1, it is shown that a perturbation analysis can account for the effects of ultrafiltration and urea generation. This permits an analytical solution of the equations which describe the variation in solute concentration in each of the compartments, which may be compared with the solution obtained when urea generation and ultrafiltration are neglected. For a typical adult patient with urea distribution volume in the range 25-40 l undergoing high-efficiency haemodialysis, the analysis suggests that the contribution of ultrafiltration to the variation in urea concentration during dialysis is similar in magnitude to the experimental errors in measuring the blood concentration of urea and that a constant volume model will be sufficiently accurate to describe urea clearance in many patients. PMID- 8719942 TI - TLD, diode and Monte Carlo dosimetry of an 192Ir source for high dose-rate brachytherapy. AB - Very few dosimetry data are available for the current generation of high-dose rate (HDR) 192Ir sources, which have broad application in remotely afterloaded brachytherapy. We have measured the two-dimensional dose rate distribution around a microSelectron-HDR source and used the results to validate Monte Carlo simulations. Thermoluminescent dosimeters (TLDs) in solid-water phantoms were used to measure the transverse-axis dose rates in the distance range 0.5-10 cm and the polar dose-rate profiles at 1.5, 3 and 5 cm distance from the source. At close distances, 2-40 mm from the HDR source, we performed transverse axis dose rate measurements with a Si diode in water. We performed diode measurements at the same distances also for a pulsed dose-rate (PDR) source to compare the results for 192Ir sources with different encapsulation. Both the HDR and the PDR sources were decayed, separated from their cables and calibrated prior to the measurements. The measured dose rates were compared with Monte Carlo photon transport calculations, which realistically modelled the experimental and source geometry at each measurement point. Agreement between Monte Carlo photon transport absolute dose-rate calculations and measurements was, on average, within 5%. From the transverse-axis experimental data, we deduced a value for the dose-rate constant lambda 0 of 192Ir HDR sources of 1.14 cGy h-1 U-1 +/- 5%. This value agrees within the experimental error with the Monte Carlo estimate of 1.115 cGy h-1 U-1 +/- 0.5%. Excellent agreement with previously measured anisotropy functions was observed. Higher anisotropy is observed for the point at 0 degree along the source cable for which no previous data have been reported. PMID- 8719943 TI - A mathematical model for predicting the temperature distribution in laser-induced hyperthermia. Experimental evaluation and applications. AB - A time-dependent mathematical model for the heat transfer in laser-induced hyperthermia has been developed. The model calculates the temperature distribution in surface-irradiated tissues. Good agreement was found between the predictions of the model and in vitro experimental results obtained for bovine liver irradiated with an expanded beam from a Nd:YAG laser. Surface evaporation of water was included in the model and experimentally verified. The discrepancy between the measured and the calculated rise in temperature at three different depths on the axis of symmetry of the irradiating beam was found to be less than 5% after 15 min of irradiation. When irradiating in air and not accounting for the surface evaporation in the model, the accuracy of the model predictions was only 75-80%. The model was then used to investigate the influence of surface evaporation of water on the total temperature distribution theoretically in a clinically relevant case. From the numerical simulations, it was shown that, simply by providing a moistened liver surface, the maximum steady-state temperature could be forced into the tissue to a depth of 4 mm. It was also shown that, by employing the numerical model during the initial phase of hyperthermia treatment, overshooting of the temperature during the transient thermal build-up time could be prevented. PMID- 8719944 TI - A filtered backprojection algorithm for axial head motion correction in fan-beam SPECT. AB - In this study we present an approximate, but practical, three-dimensional filtered backprojection (FBP) reconstruction algorithm in fan-beam SPECT to correct for axial motion (both translation and rotation). A one-dimensional filter kernel was applied to the projections. It is assumed that the object is rigid and that its axial motion can be characterized by three components: one dimensional translation and yaw and pitch rotations. It is further assumed that the motions that have occurred during the SPECT acquisition have been determined separately. The determined angular-view-dependent translation/rotation parameters were incorporated into the proposed FBP algorithm to correct for multiple axial head motions. The proposed axial head motion correction algorithm was evaluated using simulated three-dimensional Hoffman brain phantom data. Projections both with axial translation and with axial rotation, and with their combinations were generated. Images of a Hoffman brain phantom reconstructed using the proposed FBP algorithm and the conventional FBP algorithm were compared. Artefacts were observed in images without motion correction, but the artefacts were greatly reduced using the proposed reconstruction algorithm. PMID- 8719945 TI - Radiation dosimetry in human bone using electron paramagnetic resonance. AB - Accurate measurements of dose in bone are required in order to improve the dosimetry of systemic radiotherapy for osseous metastases. Bone is an integrating dosimeter which records the radiation history of the skeleton. During irradiation, electrons become trapped in the crystalline component of bone mineral (hydroxyapatite). The traps are very stable; at room temperature, emptying of the traps occurs with a half-life of many years. The population of trapped unpaired electrons is proportional to the radiation dose administered to the bone and can be measured in excised bone samples using electron paramagnetic resonance (EPR). EPR spectra of synthetic hydroxyapatite, irradiated with Co-60, were obtained at room temperature and at 77 K. At room temperature, the radiation induced signal, with a g-value of 2.001 +/- 0.001, increased linearly with absorbed dose above a lower threshold of 3 Gy, up to doses of 200 Gy. In contrast with pure hydroxyapatite, EPR spectra of excised human bone showed a broad "native' signal, due to the organic component of bone, which masks the dosimetrically important signal. This native signal is highly variable from sample to sample and precludes the use of EPR as an absolute dosimetry technique. However, after subtraction of the background signal, irradiated human bone showed a linear response with a lower limit of measurement similar to that of synthetic hydroxyapatite. Bone is an in vivo linear dosimeter which can be exploited to develop accurate estimates of the radiation dose delivered during systemic radiotherapy and teletherapy. However, improved sensitivity of the EPR dosimetry technique is necessary before it can be applied reliably in clinical situations. PMID- 8719946 TI - A dynamic phantom brain model for near-infrared spectroscopy. AB - This report describes the construction, fluid dynamics and optical properties of an in vitro model of the neonatal brain for testing near-infrared spectroscopy (NIRS) instruments. The brain model is a solid plastic structure containing a vascular network perfused with blood equilibrated with O2, N2 and CO2 in a closed circuit. The oxygenation state and haemoglobin concentration of the perfusate can be regulated and measured with a co-oximeter, providing a means to compare NIRS measurements of oxy-, deoxy- and total haemoglobin concentrations and haemoglobin O2 directly with a validated standard method. Fluid dynamic experiments revealed that the model's vasculature remains stable over time with minimal haemolysis. The model's optical properties were characterized by time-resolved and continuous wave NIRS between 670 and 850 nm as perfusate saturation was varied in the range 0-100%. Optical properties of the neonatal piglet brain were also determined by similar methods. No significant differences were found between the model and piglet brain in absorption coefficients, reduced scattering coefficients and optical pathlengths, indicating that the model optically simulates the piglet brain over a wide range of oxygenation states. These results demonstrate the potential utility of this dynamic phantom brain for testing NIRS instruments for accuracy and reliability. PMID- 8719947 TI - The effect of overlying tissue on the spatial sensitivity profile of near infrared spectroscopy. AB - The exact volume of brain tissue interrogated in near-infrared spectroscopy (NIRS) studies of cerebral oxygenation in unknown, the inhomogeneity of the head and resulting variation in optical paths making determination of the interrogated volume difficult. In this study, the spatial sensitivity profiles in simple two component inhomogeneous cylindrical models are predicted by Monte Carlo simulation in order to reveal the effects of the overlying tissue on the volume of tissue interrogated by NIRS. The predictions are validated by experimental measurements on solid "tissue equivalent' cylindrical phantoms. The phantom diameter is 30 mm and the thickness of the outer layer is 2 mm. The optical properties of the inner cylinder match those of adult brain white matter (mu' s1 = 6.4 mm-1 and mu' a1 = 0.02 mm-1), but those of the outer layer are changed (mu' s0 from 1.7 to 8.3 mm-1 and mu' a0 from 0.002 to 0.1 mm-1). Results show that the spatial sensitivity profile is largely confined to the outer layer at small optode spacing (< 15 degrees) and to the inner layer at large spacing (> 120 degrees). At intermediate angles, the sensitivity profiles are sensitive to the optical properties of the outer layer. A low mu' s0 or mu' a0 moves the profile toward the surface, at high mu' a0 it moves in towards the inner layer and at high mu' a0 it is similar to the homogeneous case. PMID- 8719948 TI - "Adaptive" changes in the behaviour of parasitized animals: a critical review. AB - Changes in host behaviour following infection with parasites are frequently reported in the literature, and are often hypothesized to be adaptive for either host or parasite. However, investigators of such phenomena often use the "adaptation" label for host behavioural changes based on their intuition and not on rigorous criteria. Alterations in host behaviour following infection can only be considered adaptive if they satisfy certain conditions: (1) they must be complex; (2) they must show signs of a purposive design; (3) they are more likely to be adaptations if they have arisen independently in several lineages of hosts or parasites; and (4) they must be shown to increase the fitness of either the host or the parasite. A survey of published examples of host behavioural changes indicates that while some are spectacularly complex and are extremely well-fitted to their presumed function, most are simple increases or decreases in an activity performed prior to infection. There are some suggestions of convergent evolution in behavioural change in distantly related host or parasite groups but more evidence is needed. Finally, most known behavioural changes have not been demonstrated to lead to fitness gains in either hosts or parasites. Few known examples satisfy more than two of the above criteria, and, in general, the adaptive function of changes in host behaviour following infection is in need of more solid proof. PMID- 8719949 TI - Characterization of a novel non-muscle myosin-related protein from Onchocerca gibsoni. AB - A cDNA library was constructed in lambda gt11 using poly(A)+ mRNA from early larvae of Onchocerca gibsoni. Screening of the library using serum from a single onchocerciasis patient yielded several strongly immunoreactive clones, one of which (OGK2) was found to encode a novel myosin-related protein. cDNA clone OGK2 contained an insert of 2017 bp, consisting of continuous open reading frame in frame with the vector, hence this clone encodes 671 amino acid residues of a larger protein. A fragment (619 nt) of the OGK2 cDNA was subcloned into the expression vector pGEX-1N to generate a glutathione S-transferase fusion protein. Polyclonal antiserum raised to this fusion protein strongly recognised an O. gibsoni protein of approximately 220 kDa. Immunolocalization studies indicated that this protein was associated predominantly with the hypodermis and a number of other specific membrane layers in the adult parasite. Myosin-related proteins are frequently immunodominant parasite antigens and in a number of studies have been shown to confer a degree of protective immunity against the corresponding parasite. Evaluation of the protective potential of the OGK2 protein, therefore, appears to be warranted. PMID- 8719950 TI - Trichinella spiralis and Trichinella pseudospiralis induce collagen synthesis by host fibroblasts in vitro and in vivo. AB - Immunoperoxidase staining of muscle infected with Trichinella spiralis for murine collagen types I and IV provided both qualitative and quantitative evidence of extensive synthesis of both types of collagen by fibroblasts in infected muscle compared to that seen in uninfected muscle. Moreover, fibroblasts in muscle infected with T. pseudospiralis, a nonencapsulating species, showed significantly less staining for both types of collagen compared to muscle from mice infected with T. spiralis. Analysis of collagen composition of isolated nurse cells using an ELISA specific for either type I or type IV murine collagen suggested that of these 2 types of collagen, only type IV basement membrane collagen is found in Trichinella capsular collagen. Excretory/secretory products of T. spiralis and T. pseudospiralis induced extensive synthesis of exclusively type IV collagen by 3T3 murine fibroblasts in vitro. PMID- 8719951 TI - Morphological and morphometric study of crustacean parasites within the genus Lernaeocera. AB - Two species of Lernaeocera are present in the southeastern North Sea. Lernaeocera lusci infects bib Trisopterus luscus, dragonet Callionymus lyra and sand goby Pomatoschistus minutas. L. minuta is a junior synonym of L. lusci. The second valid species, L. branchialis, infects whiting Merlangius merlangus. The two species can be morphologically separated by the antennary processes, which are present in L. lusci and absent in L. branchialis. Discriminant functions allow complete separation between L. lusci and L. branchialis. There is high intraspecific, host-dependent variability within L. lusci. Length of L. lusci is significantly influenced by host size, and body form is influenced by the site of attachment of L. lusci on at least one host (bib). It is suggested that L. lusci consists of 3 forms: f. lusci, f. minuta and f. lyra. PMID- 8719952 TI - Genetic evidence against a morphologically suggestive conspecificity of Dermacentor reticulatus and D. marginatus (Acari:Ixodidae). AB - International Journal for Parasitology 25: 1413-1419. Dermacentor reticulatus and D. marginatus exhibit overlapping phenotypes. The possibility of conspecificity was investigated on the nucleotide level by comparing DNA sequences of the second internal transcribed spacer (ITS 2) of the rDNA gene. The inter-specific polymorphism was more than 20-times greater than the intra-specific polymorphism of 3 D. reticulatus strains of different geographic origins. Furthermore, the degree of polymorphisms between D. reticulatus and D. marginatus was found to be of the same order of magnitude as that between D. andersoni and D. variabilis, for which separate species status is accepted. These genomic findings do not support a possible conspecificity of D. reticulatus and D. marginatus. PMID- 8719953 TI - Cryopreservation of the first-stage larvae of trichostrongylid nematode parasites. AB - First stage (L1) larvae of Haemonchus contortus, Trichostrongylus colubriformis and Ostertagia circumcincta can be cryopreserved in the presence of DMSO using a two-step freezing protocol involving an initial period at -80 degrees C prior to transfer to liquid nitrogen. Thawed L1 larvae continue development in vitro producing third stage (L3) larvae that are infective to sheep when dosed per os. Establishment rates for L3 larvae grown from thawed L1 larvae were 40 and 80% for H. contortus and T. colubriformis, respectively. There was no difference in survival or infectivity between benzimidazole (BZ)-susceptible and BZ-resistant H. contortus isolates and cryopreservation caused no shift in their BZ-resistance status as indicated in an in vitro larval development assay. Cryopreservation also had no effect on the sensitivity of these isolates to the avermectins or levamisole in vitro. High survival rates (60-70%), good levels of establishment and the stability of anthelmintic resistance status of isolates indicate that little if any selection occurs during the cryopreservation process. L1 larvae of all 3 species have been successfully recovered after 16 months storage in liquid nitrogen, cultured to the L3 stage and established in sheep. PMID- 8719954 TI - Ultrastructure of spermiogenesis and the mature spermatozoon of Tetrabothrius erostris Loennberg, 1896 (Cestoda, Tetrabothriidae). AB - The spermiogenesis of Tetrabothrius erostris is characterized by the following events: formation of a differentiation zone containing 2 basal bodies and a pair of rootlets; one of the basal bodies gives rise to a free flagellum, the other induces formation of a flagellar bud; rotation at 90 degrees of the flagellum prior to its fusion with the middle cytoplasmic process of the differentiation zone and partial rotation of the flagellar bud; penetration of the nucleus between the rootlets and appearance of a spur-like protrusion in the differentiation zone; elongation and twisting of the differentiation zone, resulting in twisting of the peripheral microtubules and migration of the nucleus; formation of a crested body; proximal densification of the spermatozoon prior to its detachment from the spermatid rosette. The mature spermatozoon has a single axoneme of 9+"1" type and twisted peripheral microtubules. It consists of 3 portions: a proximal part with a crested body, a middle region rich in beta glycogen, and a distal part containing the nucleus. The pattern of spermiogenesis resembles most closely that in phyllobothriid tetraphyllideans, and probably reflects a relationship of the family Tetrabothriidae with this group. PMID- 8719955 TI - Development of an injectable formulation of albendazole and in vivo evaluation of its efficacy against Echinococcus multilocularis metacestode. AB - The loading of poly (D, L-lactide) nanoparticles with ABZ has led us to evaluate the potential of this new colloidal drug delivery system against E. multilocularis, using a murine model of hepatic alveolar echinococcosis. ABZ loaded nanoparticles had a monodisperse size distribution between 100 and 150 nm. The efficiency of drug loading to nanoparticles was over 97%. In vitro, at an ABZ concentration of 1.0 microgram ml-1, the formulation had no toxicity for peritoneal macrophages harvested from uninfected mice. In vivo, the ABZ-loaded nanoparticles exhibited no signs of toxicity at any of the doses tested. Intravenous injections of 6 mg kg-1 of bound ABZ to infected mice had an equivalent antiparasitic effect on the metacestode growth to that of a treatment with 1500 mg kg-1 of orally administered free ABZ. The parasite hepatic superficial size as well as the peritoneal metastatic burden was significantly reduced by these 2 courses of treatment, as compared to those of untreated mice. Our results should encourage further study in order to explain the absence of dose-dependent efficacy of ABZ-loaded nanoparticles demonstrated in the present study. PMID- 8719956 TI - Evaluation of the efficacy of albendazole against the larvae of Taenia solium in experimentally infected pigs, and kinetics of the immune response. AB - Cysticercosis, a disease of economic and public health importance, is caused by Cysticercus cellulosae, the metacestode stage of Taenia solium. Experimental induction of cysticercosis was achieved in young pigs by feeding an optimum dose of 20,000 T. solium (Indian strain) eggs after immunosuppression, to assess the effect of albendazole and development of the immune response to cysticercus antigens before and after treatment. Histopathological studies revealed the presence of cysticerei in liver, lungs and muscles. Treatment with albendazole at 15 mg kg-1 body weight daily for 30 days starting from day 0 or 15 days post infection resulted in 100% cure rates. Increases in antibody titre to crude soluble extract and a Sephadek G-200 purified antigenic fraction of Cysticercus cellulosae were found on days 25, 40 and 55 post-infection in untreated pigs and those in which treatment started on day 15 post-infection, whereas no increase in antibody response was observed in pigs in which treatment started on day 0. PMID- 8719957 TI - Kinetic disposition of an emulsifiable concentrate formulation of deltamethrin applied to sheep in a plunge-dip and its effect on lice. AB - An emulsifiable concentrate formulation of the synthetic pyrethroid insecticide deltamethrin was applied in a plunge dip, 3 weeks after shearing, to a group of 5 Merino sheep infested with sheep body lice, Bovicola ovis. Deltamethrin concentrations on the wool were measured at regular intervals between 1 and 98 days after treatment and were not significantly different (P > 0.05) between sites on the dorsal mid-line, upper or lower flank. Levels in the tip of the fleece were significantly greater than those in the base, indicating that there was little movement of deltamethrin down the staple as the wool grew. Most lice were killed after 20 h of exposure in vitro to wool samples collected between 1 and 28 days after treatment. However, many lice survived in samples containing a similar concentration of deltamethrin, but collected between 35 and 98 days after treatment. Numbers of lice surviving increased with the sampling time after treatment, suggesting that the bioavailability of the deltamethrin changed as the insecticide aged in the fleece. Some transfer of deltamethrin occurred from treated to untreated sheep. The levels of deltamethrin were higher in sheep placed in contact with the treated group at 14 days after treatment than in those which were in contact from 43 days after treatment. PMID- 8719958 TI - Antimalarial properties of soy-bean fat emulsions. AB - Intralipid and Ivelip are commercial preparations of soy-bean lipid extracts used for intravenous supplementation of lipids in various clinical conditions. They were found to inhibit the growth of Plasmodium falciparum in culture with an IC50 of 8.07 +/- 2.13 and 13.32 +/- 2.05 mg.ml-1, respectively. Intralipid rapidly and efficiently inhibited nucleic acid synthesis in cultured P. falciparum, exhibiting full inhibitory activity in less than 2 h. Ivelip injected intraperitoneally, was found by the 4-day suppressive test to be active in vivo against P. vinckei petteri within the normal recommended regimen for dietary lipid supply (0.5-4 g.kg-1), but it was impossible to obtain a radical cure even with very high doses (6.4 g.kg-1). Ivelip was less effective against P. berghei and P. yoelii nigeriensis. As Ivelip showed no interference with the antimalarial activity of chloroquine, it could be considered for use in the treatment of severe human malaria in association with 4-aminoquinolines to expedite the clearance of parasites. PMID- 8719959 TI - Cytopathology and release of an RNA virus from a strain of Trichomonas vaginalis. AB - A strain of Trichomonas vaginalis infected with a double-stranded RNA virus showed pronounced cytopathology in the form of giant syncytia generated by the recruitment of single cells. The giant cells ultimately lysed, releasing virus into the culture medium. In the infected cells, clusters of electron-dense particles resembling viral structures were found in the cytoplasm. In addition, distinctive inclusions composed of similar particles were present in the nuclei of some cells. Double-stranded viral RNA of 5.5 kbp was demonstrated in both cytoplasmic and nuclear fractions from these cells. Viral particles collected from the cell-free culture supernatant were of the same shape and size as the RNA virus isolated from a strain of T. vaginalis described previously (Wang & Wang, Journal of Biological Chemistry, 260: 3697-3702, 1985; Wang & Wang, Proceedings of the National Academy of Sciences of the U.S.A. 83: 7956-7960, 1986) which does not show this cytopathology. PMID- 8719960 TI - Influence of hydrogen peroxide on acid-fast staining of Cryptosporidium parvum oocysts. AB - Infections by the protozoan parasite Cryptosporidium parvum are routinely diagnosed by modified Ziehl-Neelsen (acid-fast) staining of faecal preparations despite the counterstaining and ghost-like appearance of some oocysts. Quantitative studies demonstrated that only a small percentage of oocysts excreted by naturally infected newborn calves displayed acid-fast characteristics, but that percentage increased when the time between excretion and sample staining was increased. The treatment of faecal samples with hydrogen peroxide (10 min, 5 vol. final concentration) caused all oocysts to become acid fast, with up to 40-fold increases in test sensitivity in samples treated and stained within 3 h of excretion. Flow-cytometry analysis of hydrogen peroxide treated oocysts also demonstrated increased labelling of oocysts by a commercial monoclonal antibody preparation commonly used for diagnosis. PMID- 8719961 TI - A model for nematodiasis in New Zealand lambs: the effect of drenching regime and grazing management on the development of anthelmintic resistance. AB - A model for nematodiasis in lambs was expanded to incorporate both the contribution of ewes to nematode epidemiology and the genetic parameters required to simulate the development of anthelmintic resistance in the nematode population. The expanded model was used to assess the impact of various drench and grazing management strategies for ewes and lambs on the rate of development of anthelmintic resistance. Three grazing management options, under a range of drenching schedules, were compared: one in which lambs and ewes were rotationally grazed as separate flocks over the same area after weaning (common grazing); a second in which lambs were grazed, after weaning, on areas from which ewes were excluded (separate grazing); and a third in which lambs were moved to "safe" pasture at weaning and again in early autumn (integrated control). Drenching strategies examined under the first 2 grazing options included a 5 lamb-drench "preventive" programme with 0, 1, 2, 3 or 4 additional lamb drenches, and 0 or 1 ewe drench treatment at either tall-docking or mating. Under the third grazing option, lambs were given either 1 or 2 drench treatments at or following each move to safe pasture and ewes 0 or 1 drench treatment at either tail-docking (i.e., 3-4 weeks after lambing) or mating. Model output suggests that drenching ewes prior to any lamb drenching programme is likely to significantly increase selection for drench resistance by pre-selecting the larval challenge to the lambs and, under some grazing systems, by reducing the diluting effect of eggs of susceptible genotypes passed by undrenched ewes. The results highlight the potential importance of undrenched ewes as a refuge for susceptible worm genotypes and indicate that on its own, drenching frequency is likely to be a poor indicator of selection pressure for resistance and thus of limited value in selecting strategies for the management of anthelmintic resistance. PMID- 8719962 TI - Experimental Oesophagostomum dentatum infection in the pig: worm populations resulting from single infections with three doses of larvae. AB - This report describes the effect of different dose levels of infection upon worm burdens and development and fecundity of the parasites. Three groups each of 40, 9-week-old, helminth naive pigs were inoculated once with either 2000 (group A), 20,000 (group B), or 200,000 (group C) infective third stage larvae of Oesophagostomum dentatum. Subgroups of 5 pigs from each major group were killed 3, 6, 11, 14, 18, 25, 34 and 47 days post inoculation (p.i.) and the large intestinal worm burdens were determined. Faecal egg counts were determined at frequent intervals after day 13 p.i. There were no overt clinical signs of gastrointestinal helminthosis during the experiment. Faecal egg counts became positive in groups A and B at around day 19 p.i., whereas most pigs in the high dose group C did not have positive egg counts until day 27-33 p.i. and some pigs remained with zero egg counts until the end of the study. Throughout the experiment the worm populations in group C consisted mainly of immature larval stages, while those in groups A and B were predominantly adult stages after days 14-18. Adult worms from the low dose group A were significantly longer than those from group C. At high population densities, stunted development of worms and reduced fecundity among female worms were found. Furthermore, there was a tendency for the distribution of the worms within the intestine to be altered with increasing population size. PMID- 8719963 TI - Comparison of local and systemic responsiveness of lymphocytes in vitro to Bovicola ovis antigen and concanavalin A in B. ovis-infested and naive lambs. AB - The in vitro proliferation assay was used to determine lymphocyte responsiveness to soluble antigen of B. ovis and to Concanavalin A (Con A) in peripheral blood, spleen and various lymph nodes from B. ovis-infested and naive lambs. From March to July, an assay of monthly blood samples showed generally higher proliferative responses to antigen and Con A in B. ovis-infested than naive lambs. The proliferative response of cells from the skin-draining prescapular lymph nodes to B. ovis antigen was significantly higher in B. ovis-infested than naive lambs. Responses of cells from the medial iliac, mediastinal and mesenteric lymph nodes (which do not receive lymph from the skin) and spleen showed no significant differences between groups. Within the B. ovis-infested lambs, the response of cells from the prescapular lymph node was significantly higher than that from any other lymphoid organ examined. Responsiveness of the prescapular, medial iliac and mesenteric lymph node and spleen cells to Con A was not significantly different between groups, while mediastinal lymph node cells showed a significantly higher response in B. ovis-infested lambs. The data indicate that the antigen-specific cellular immune response is operating mainly locally, at the level of the skin and draining lymph nodes. Responses to the T cell mitogen Con A did not support non-specific immunodepression as reported in other ectoparasite/host systems. PMID- 8719964 TI - Use of a monoclonal antibody to ovine IgE for fly strike studies in sheep. AB - A new monoclonal raised against sheep IgE was used to examine sera and wound exudates from sheep which had been struck by Lucilia cuprina in the field. The antibody was also used to detect the presence of IgE in sera and skin sections from sheep which had been artificially infected with fly larvae 3 times. Neither total, nor L. cuprina specific circulating IgE could be detected in serum or wound exudates from struck sheep. Cell bound IgE was, however, identified by the monoclonal in skin sections from struck sheep and from a control sheep which had not been struck. No difference in the number of IgE positive cells was observed between the control and 2 of the 3 artificially infected sheep, and none of the latter showed an increase in IgE positive cells even after 3 infections. One sheep showed twice as many IgE positive cells as the other treated sheep and the third larval infection was difficult to establish and limited in size and severity. This suggests a relationship between innate resistance to strike and the number of IgE positive cells present in skin. PMID- 8719966 TI - Biochemical properties of purified cathepsin B from Schistosoma mansoni. AB - A previously described "major acidic proteinase" of adult Schistosoma mansoni, believed to play a key role in the parasite's metabolism, has been identified as a cathepsin B (Sm31). Purified Sm cathepsin B was not recognized by anti-Sm32 or anti-cathepsin L antibodies. The enzyme hydrolyzes the synthetic protease substrates Z-Arg-Arg-AMC and Z-Phe-Arg-AMC as well as protein substrates. Its pH optimum is 3.0 with serum albumin, 4.0-5.0 with globin and 5.5-6.0 with the synthetic substrates. The enzyme was inactivated by cysteine proteinase inhibitors. Its activity against protein substrates would support the hypothesis that it plays a role in schistosome nutrition. PMID- 8719965 TI - ITS2 ribosomal RNA indicates Schistosoma hippopotami is a distinct species. AB - The internal transcribed spacer region of the ribosomal RNA, ITS2, was sequenced from a single specimen of S. hippopotami collected from a pulmonary artery of the hippopotamus, Hippopotamus amphibius in South Africa. The nucleotide sequence was aligned with those of S. mansoni, S. rodhaini, S. haematobium, S. intercalatum, S. curassoni, S. bovis and S. japonicum. Both maximum parsimony and genetic distance analyses were performed on these data sets. Using S. japonicum as outgroup to the African schistosomes, a single most-parasmonious tree was obtained of length 64 steps with a consistency index of 1-S. hippopotami was the sister-group to the remaining African species. This species has lateral-spined eggs and its basal position in the tree suggests that this condition is primitive and that terminal-spined eggs developed secondarily. Molecular data clearly show that S. hippopotami cannot be considered synonymous with S. mansoni. Assuming the hippopotamus is the normal host of S. hippopotami, phylogenetic analysis is consistent with an ancient association between schistosomes and ungulates. PMID- 8719967 TI - Alpha 2-adrenoceptors and I2 sites in the mammalian central nervous system. AB - The presence of alpha 2-adrenoceptors and I2 sites in the central nervous system is well established. This review provides a complete summary of this literature, commencing with the first description of alpha 2-adrenoceptors and I2 sites in the brain and covering in detail their subclassification, distribution, and function. Furthermore, it highlights some important differences between central alpha 2-adrenoceptors and I2 sites. PMID- 8719968 TI - Interactions involving new antiepileptic drugs. AB - The advent of new antiepileptic drugs (AED) has increased the opportunities for interaction. Clinicians seek therapeutic interactions in which two AED together have greater efficacy than either drug alone; there are case reports of such, but few prospective studies. Interactions must also be suspected when the adverse effects of a new AED differ according to the co-medication. The basis can be pharmacodynamic, but more frequently it is pharmacokinetic. Inhibition of cytochrome P450 enzymes by the new drugs is more common than induction. There are important implications for the design of clinical trials and the planning of treatment changes in patients. PMID- 8719969 TI - The effects of chronic hypoxia on the pharmacological responsiveness of the pulmonary artery. AB - Chronic hypoxia (CH) is associated with several cardiopulmonary disorders. In vitro and in vivo studies have established the morphological changes, but yielded conflicting results about the functional changes induced by CH in the pulmonary vascular bed. CH increases the responsiveness to endothelium-dependent vasodilators in perfused lungs; however, in artery rings, a reduction is found in endothelium-dependent vasodilation. In CH, vasoconstriction induced by endothelin 1 is enhanced and vasodilation produced by atrial natriuretic peptide and K+ channel openers is increased. Vasoconstriction produced by acute hypoxia can be either enhanced or reduced by CH, depending on the experimental protocol. An understanding of the functional changes associated with CH is particularly important for a rational approach to the treatment of disorders associated with CH. PMID- 8719970 TI - Therapeutic applications of ribozymes. AB - The demonstration that RNA can be cleaved by cis or trans ribozymes (catalytic RNAs, RNA enzymes) has potentially important therapeutic implications. Since their discovery in the 1980s, the biochemistry and conserved sequences of ribozymes have been well characterized. Ribozymes are effective modulators of gene expression because of their simple structure, sitespecific cleavage activity, and catalytic potential. The targets of ribozyme-mediated gene modulation have ranged from cancer cells to foreign genes that cause infectious diseases. Additional target sites for ribozymes are in initial phases of development and design. Ribozymes have been targeted against a myriad of genes, including oncogenes (ras, BCR-ABL, c-fos) and drug resistance genes, as well as the human immunodeficiency virus-type I genome. These ribozymes have cleaved the target RNAs in vitro and altered the cellular pathology. Currently, the therapeutic application of ribozymes to human diseases is limited by gene transfer systems. It is anticipated that ribozymes ultimately will play an important role in human gene therapy. PMID- 8719971 TI - Molecular, cellular, and clinical aspects of the pharmacology of 20(S)camptothecin and its derivatives. AB - The discovery of the plant alkaloid 20(S)camptothecin (CPT), which displayed potent antitumor activity in preclinical trials, has led to the identification of a novel target of cancer chemotherapy: the nuclear enzyme topoisomerase I. The mechanism by which CPT induces cytotoxicity is the topic of continued research, but appears to be mediated by the stabilisation of transient "cleavable" topoisomerase I-DNA complexes. The pharmacology of CPT and its derivatives is complicated by the apparent requirement of an alpha-hydroxy-delta-lactone ring, which, unfortunately, is hydrolysed reversibly to form inactive carboxylates. Recent research has shown that the extent of hydrolysis in vivo varies between the various derivatives and that this may be an important factor in determining antitumoral activity. In this review, we discuss recent developments in our understanding of the molecular, cellular, and clinical pharmacology of CPT and several of the more promising derivatives. PMID- 8719972 TI - Central noradrenergic neurones and stress. AB - This review discusses evidence that central noradrenergic neurones are involved in the response and adaptation to stress. Stress-induced changes in neurochemical processes affecting neurotransmission are detailed first, followed by evidence that these are modified by psychotropic agents. The possibility that these changes influence behaviour is also reviewed. Collectively, evidence suggests that these neurones form a sophisticated network in which the effects of stress differ between brain regions and type of stress imposed. Presynaptic processes governing transmitter release, and the net activation of different adrenoceptor subtypes all contribute to the impact of central noradrenergic neurones on the behavioural response to stress. PMID- 8719973 TI - Traditional healers in Tanzania: the perception of malaria and its causes. AB - The coordination of traditional and western medicine is still in its infancy in most African countries. Although there is much discussion about the contribution of traditional medicine and its practitioners, especially on the primary health care level, it has rarely be done in practice. This is probably due to the lack of knowledge of how to do it, because a serious attempt to include traditional medicine in health planning would presuppose that it is known what traditional medicine has specifically to offer for certain diseases/illnesses and how traditional healers manage such conditions. The aim of this study was to investigate the management of malaria by traditional healers in different areas in Tanzania. This included looking at the perception, the causation concepts and the knowledge about prevention of the disease/illness of malaria. For this purpose traditional healers were interviewed in different rural and urban places in Tanzania: in the Kilombero valley (Kilombero/Ulanga District), on the main island of Ukerewe (Ukerewe District), in the region near Bukoba town (Bukoba District) and in the settlement of Dar es Salaam (largest town of Tanzania). The results of the study show that most of the interviewed traditional healers were very familiar with the signs and symptoms relating to malaria, as it is defined by western medicine. Many healers were aware of different manifestations of malaria and attributed to them different local names, which match the scientific terms which describe the different types of Plasmodium falciparum malaria, such as cerebral malaria, clinical malaria or febrile type, and gastrointestinal type, respectively. Differences compared to western medical knowledge were found for concepts of causation, and in the fact that severe malaria in children may not be perceived as being associated with malaria. PMID- 8719974 TI - Traditional healers in Tanzania: the treatment of malaria with plant remedies. AB - In order to collect ethnobotanical information about antimalarial plants which is essential for the further evaluation of the efficacy of plants an antimalarial remedies, we investigated the management of malaria with traditional herbal remedies, including the use, preparation and administration, by traditional healers in Tanzania. Interviews with traditional healers were conducted in different rural and urban places in Tanzania: in the Kilombero valley (Kilombero/Ulanga District), on the main island of Ukerewe (Ukerewe District), in the region near Bukoba town (Bukoba district), and in the settlement of Dar es Salaam (largest city in Tanzania). The results of the study show that all traditional healers treat malaria with herbal remedies consisting of one to five different plants. The list of plants which they use for antimalarial treatment contains a large number of species from different families. Multiple citations of plants by different healers were rare. Most of the respondents attributed to the plants mentioned, or to the remedies made from them, specific effects and sometimes side effects, explaining and illustrating their use or non-use for different patients or manifestations of the disease/illness. PMID- 8719975 TI - Traditional healers in Tanzania: sociocultural profile and three short portraits. AB - Traditional healers are an important part of African societies, but unfortunately the knowledge of the extent and character of traditional healing and the people involved in the practice is limited and impressionistic. They are frequently ignored in studies of user/provider patterns, although they cover the health needs of a substantial proportion of the population. For future health planning it is necessary to know what the reasons are that even in big cities, where western health care services are available, traditional healers flourish, and even compete with each other for certain aspects. The aim of this study was to investigate certain aspects of the profession of traditional healing in general in different areas in Tanzania in order to get an idea about the kind of traditional medical services which are available, and about the people who provide such services. For this reason traditional healers were interviewed with a semi-structured questionnaire in different rural and urban places: in the Kilombero valley (Kilombero/Ulanga district), on the main island of Ukerewe (Ukerewe District), and in the region near Bukoba town (Bukoba District), and in the settlement of Dar es Salaam (largest city of Tanzania). The results of the study show that traditional healers are a very heterogeneous group of persons not having much in common relating to their religion, sex and level of education. The traditional practice is very often taken over from a family member, but also other reasons for becoming a healer, like initiation through ancestor spirits, are very frequently given. More than 50% of the respondents practice full time. These full time practitioners are mainly found among men and in the younger age group. Treatment of in-patients, who can stay in special patient-houses, is offered by half of the traditional healers. Divination used as a diagnostic tool was found mainly among men. Referral of patients to the hospital was mentioned by almost all respondents in cases where they failed with their own treatment or when they knew that the patient would be better treated in the hospital or dispensary. PMID- 8719976 TI - Anthelmintic activity of papaya latex against patent Heligmosomoides polygyrus infections in mice. AB - The purpose of this experiment was to study the possible anthelmintic activity of papaya latex (Carica papaya) against Heligmosomoides polygyrus in experimentally infected mice. Five groups of BALB/C mice were infected with 100 Heligmosomoides polygyrus infective larvae/mouse. After patency (day 22) four groups of mice (groups B, C, D and E) were given papaya latex suspended in water at dose levels of 2, 4, 6 and 8 g of papaya latex/kg body weight, respectively. One group of mice (group A) served as non-treated controls. All animals were necropsied on day 25, i.e. 3 days after treatment, for post-mortem worm counts. The papaya latex showed an antiparasitic efficacy of 55.5, 60.3, 67.9 and 84.5% in groups B, C, D and E, respectively. The results may suggest a potential role of papaya latex as an anthelmintic against patent intestinal nematodes of mammalian hosts. PMID- 8719978 TI - Fluorodeoxyglucose (FDG) and tumour metabolism. PMID- 8719977 TI - Superoxide and traditional Chinese medicines. AB - In traditional Chinese medicinal practices, herbs are classified as 'cold', 'neutral', or 'hot'. Fluorometric analysis of herbs with 'cold' properties revealed that these herbs produce large amounts of superoxide. In contrast, herbs with 'hot' properties have scavenging activities. We believe that this electron transfer to form superoxide and the scavenging of superoxide may elucidate the phenomena of the 'yin' (represented by 'cold') and 'yang' (represented by 'hot') respectively. PMID- 8719979 TI - Radiolabelled human immunoglobulins. AB - Radiolabelled human immunoglobulins have been available for 5 years. Within that time there have been many publications on the use of these agents labelled with both technetium-99m (99Tcm) and indium-111 (111In). The results of these data appear contradictory and only now do we understand both some of the mechanisms of radiolabelled immunoglobulins and some of their limitations. While 111In polyclonal immunoglobulin seems to have a wide range of clinical applications in infection and inflammation, the more readily available 99Tcm-labelled product is best suited to the localization of peripheral bone and joint disease. The ease of producing these agents from commercially available immunoglobulins will ensure continued interest and research into these new tracers. PMID- 8719980 TI - Clinical significance of decreased myocardial uptake of 123I-BMIPP in patients with stable effort angina pectoris. AB - The aim of this study was to assess the feasibility of resting myocardial fatty acid metabolic imaging with 123I-beta-methyliodophenyl-pentadecanoic acid (123I BMIPP) for the detection of patients with stable effort angina pectoris and to clarify the clinical significance of abnormal 123I-BMIPP images. Myocardial imaging with 123I-BMIPP at rest and 99Tcm-methoxyisobutyl isonitrile (99Tcm-MIBI) at rest and during treadmill exercise was performed in 46 patients with suspected effort angina pectoris. Resting 123I-BMIPP imaging detected 43% (17/40) of patients with significant (> or = 50%) coronary artery stenosis and 59% (17/29) of patients with exercise-induced myocardial ischaemia. The patients with abnormal 123I-BMIPP images terminated exercise after a shorter period (4.5 +/- 2.6 vs 6.7 +/- 4.1 min; P < 0.01) and at a lower rate pressure product (16,124 +/ 5211 vs 20,246 +/- 6564 mmHg x beats min-1; P < 0.01) than those with normal 123I-BMIPP images. The presence of ST depression during the exercise test (77 vs 52%; P < 0.05), severe coronary stenosis exceeding 90% (88 vs 43%; P < 0.01), collateral vessels (35 vs 9%; P < 0.01) and a wall motion abnormality of hypokinesis/akinesis (53 vs 30%; P < 0.05) were more frequently seen in patients with abnormal 123I-BMIPP images than in those with normal images. Resting 123I BMIPP imaging was able to detect the presence of coronary artery stenosis and exercise-induced myocardial ischaemia with moderate sensitivity, and to determine the functional severity of coronary artery disease. PMID- 8719981 TI - Diagnostic approach in acute pulmonary embolism: perfusion scintigraphy versus spiral computed tomography. AB - For diagnosing acute pulmonary embolism, perfusion scintigraphy in combination with ventilation studies is a well-established and sensitive method. Recently, computed tomography (CT) incorporating an angiographic technique was introduced for evaluating the pulmonary arterial system without breathing artefacts. The present study compared lung scans with spiral CT data to establish the diagnostic value of the latter approach. Twenty-five patients with a clinical suspicion of acute pulmonary embolism were examined using both methods. Perfusion and ventilation lung scans were done and spiral volumetric CT studies were performed. In 18 patients, pulmonary embolism was diagnosed using both methods. However, the results were interpreted differently. Particularly when centrally localized emboli were present in both pulmonary arteries, assessment of the more affected side was difficult on CT and depended on the experience of the physicians interpreting the scans. Small, (sub-)segmental emboli could not be demonstrated on CT. This may explain the false-positive and false-negative results, respectively. Spiral CT enables an exact demonstration of thrombosis in severe, centrally localized emboli. In incompletely occluded vessels, estimation of the degree and the location of perfusion defects is better with scintigraphy. Based on the present results and the technical equipment necessary for spiral CT, it is concluded that perfusion scintigraphy remains the method of choice for diagnosing acute pulmonary emboli. PMID- 8719982 TI - Brain death: the evaluation of semi-quantitative parameters and other signs in HMPAO scintigraphy. AB - The aim of this study was to facilitate the diagnosis of brain death based on semi-quantitative parameters as well as the other scintigraphic signs and symptoms of brain death. Altogether, 72 studies were performed in 54 patients (35 males, 19 females) to evaluate the diagnoses of brain death which were confirmed in 49 cases. Perfusion dynamic acquisition and static imaging were performed after the intravenous injection of 700-900 MBq 99Tcm-hexamethylpropyleneamine oxime (HMPAO). We reviewed the following parameters retrospectively: (1) qualitative visual assessment of the presence or absence of delayed HMPAO deposition in the brain area; (2) qualitative visual assessment of a 'three pronged fork' of the cerebral arteries during the first-pass study; (3) any evidence of 'hollow skull' during the first-pass study; (4) any evidence of 'hot nose'; (5) blood flow index (i.e. the ratio of the area under the brain time activity curve vs that under the curve for the carotid arteries); and (6) delayed uptake index (i.e. the ratio of HMPAO deposition in the brain vs that in the facial regions of interest). We were able to demonstrate the validity of individual signs and symptoms and to confirm the highly reliable nature of semi quantitative scintigraphic parameters. PMID- 8719983 TI - Nigrofrontal dopaminergic function as assessed by 18F-dopa PET. AB - The existence of the nigrofrontal dopaminergic pathway has been demonstrated in neuroanatomical studies. We evaluated the presynaptic nigrofrontal dopaminergic function using 18F-dopa (FD) positron emission tomography (PET). The multiple time PET data in the frontal cortex from 20 to 70 min post-injection for FD were evaluated by Patlak analysis using the cerebellar time-activity curve as an input function. The frontal FD uptake rate constants could not be determined in 5 of 12 normal volunteers because of large deviations in the plots. There were no significant differences between the subjects among whom the frontal FD uptake rate constants could or could not be determined regarding the amount of FD injected, the frontal 18F counts, or whether or not they were pretreated with carbidopa. The uptake constants were determined in 9 or 12 patients with parkinsonian syndrome. While the mean (+/- S.D.) uptake constants in patients with Parkinson's disease (2.89 +/- 0.06 x 10(-3), n = 4) and in patients with progressive supranuclear palsy (2.81 +/- 0.10 x 10(-3), n = 3) were not significantly different from those in the normal volunteers (2.93 +/- 0.14 x 10( 3)), those in two patients with corticobasal degeneration (2.42 and 2.46, respectively) decreased in comparison to the control values. Differences in the nigrofrontal presynaptic dopaminergic function as assessed by FD-PET may explain the different pathogenesis and also help to differentiate between corticobasal degeneration and other parkinsonian syndromes, such as Parkinson's disease and progressive supranuclear palsy. PMID- 8719984 TI - Can the microsphere model by applied to cerebral blood flow measurement using N isopropyl-p-[123I]iodoamphetamine with SPET regardless of washout from brain tissue? AB - The microsphere model and the continuous withdrawal of arterial blood have commonly been used in clinical studies when measuring regional cerebral blood flow (rCBF) by N-isopropyl-p-[123I]iodoamphetamine (IMP) single photon emission tomography. The method is considered to underestimate rCBF because of the washout of tracer from brain tissue; however, the extent of this underestimation is not known. To assess whether this underestimation can be determined quantitatively, we performed simulation analysis based on the microsphere model and the two compartment model [influx, K1(rCBF); and outflux, k2(washout)] using the time activity curves of 123I-IMP in arterial blood [Ca(t)] of 10 subjects. With the microsphere method, rCBF values fell as time post-injection increased. The extent of underestimation of rCBF was 4.7 +/- 0.28% (mean +/- S.D.) at 5 min, 10.2 +/- 0.42% at 10 min and 15.2 +/- 0.55% at 15 min. There was little variation in the extent of underestimation and it was not dependent on the Ca(t) of the subjects. We therefore considered the results to be generally applicable to various studies of the microsphere model. As 4.7% is considered to be negligible in clinical studies, we conclude that the microsphere model can be applied to obtain accurate measurement of rCBF up to 5 min regardless of washout. PMID- 8719985 TI - The effect of phantom wall thickness on volume determination in SPET. AB - Assessment, using phantoms, of a a three-dimensional, second-derivative, surface detection algorithm for accurately determining the volumes of structures in single photon emission tomography (SPET) has shown the performance of the algorithm to be highly dependent on phantom wall thickness. With a 30% background activity level, the volume of a 600-cm3 cylinder with wall thickness approximately 0.4 mm was measured to an accuracy of approximately 1%, whereas for an almost identical cylinder with a wall thickness of approximately 3 mm the measured volume was underestimated by about 14%. To further investigate this 'wall-thickness' effect, theoretically generated SPET data have been produced, simulating a set of low-contrast cylindrical phantoms with identical internal dimensions and wall thicknesses ranging from 0 to 6 mm. These image data have demonstrated a 10% reduction in the calculated volume for wall thicknesses as thin as 1 mm. A less acute dependence is also demonstrated for a threshold-based quantitation algorithm, where a wall thickness of 4-5 mm is required to produce an effect of similar magnitude. The underlying cause behind this 'wall-thickness' effect is undoubtedly the perturbation in the count profile across the surface boundary, which results form the cold region of the phantom wall. Thus, phantom wall thickness will have an effect on the performance of most automated quantitation techniques, both two- and three-dimensional, since the majority must incorporate some form of analysis on this count profile. PMID- 8719986 TI - A survey of dual-energy X-ray absorptiometry (DEXA) normal reference ranges used within the UK and their effect on patient classification. AB - Differences between dual-energy X-ray absorptiometry (DEXA) normal ranges can lead to patients being characterized as osteoporotic using one range and normal using another. To investigate the diversity of normal ranges used within the UK, a survey of all DEXA sites was carried out with a 60.6% response rate. The effect of the different ranges was evaluated by translating each range to an equivalent range for a Norland XR-26 system and applying the ranges to stratify a representative sample of over 1000 patients into grades of bone density based on percentages of age-matched mean BMD, Z-scores and T-scores. The effect of femoral neck and P/A spine L2-L4 regions was considered both separately and jointly. Large differences between the normal ranges were apparent, which resulted in the classification of the number of patients with a Z-score of less than -2.0 varying by a factor of more than 20 for the femoral neck and more than 3 for the spine. The number of patients defined as osteoporotic by a T-score less than -2.5 varied from none to over one-third of patients for the hip and by a factor of almost 3 for the spine. The exclusion criteria used for construction of the normal ranges varied markedly with none constructed using population-based sampling. Smoothing of normal ranges was carried out by DEXA manufacturers, while local normal ranges made use of raw unprocessed data. There is reason to question the validity of such processing. We recommend the construction of a unified UK normal range applicable to all UK DEXA systems in order to harmonize patient management and care. PMID- 8719987 TI - Home reporting for the nuclear clinician? AB - The computer-based 'home office' is becoming a widely accepted mode of operation for modern businesses. It is implausible to believe that a nuclear medicine department can be covered permanently at a distance by a single physician, but it should be possible to provide cover for colleagues during sickness or at night or weekends. We have used a 486 PC with a high-resolution screen and software provided by LINK Medical Ltd to obtain images from hospital sites using a modem link to ADAC, Bartec and Nuclear Diagnostic SUN workstations. The data were transferred via standard telephone lines to the homes of two of the authors. During a trial period lasting several months, 60 lung scans, 20 bone scans, 1 gastrointestinal bleeding study, 4 leukocyte scans, 5 bone tomograms, 9 renograms, 6 myocardial perfusion tomograms and 2 gated cardiac studies were transferred. The system allowed transfer of a 128 x 128 eight-view lung scan to be completed in approximately 2 min. The program on the PC allowed alteration of individual image contrast, image rotation, cine display and a variety of colour scales to enhance image interpretation. A system to transfer chest X-rays has been developed and typical transfer times are approximately 3.5 min. Within the viewing protocol on the PC, a reporting window was available with the ability to fax the report directly to the hospital. The system allowed consultants who live at a distance from their nuclear medicine departments to provide cover and is now used as an integral part of our out-of-hours service. The system also allows cover of satellite units or to provide cover for junior staff at night or weekends. PMID- 8719988 TI - 99Tcm-dextran: a potential inflammation-seeking radiopharmaceutical. AB - The aim of this study was to test whether 99Tcm-dextran can be used for imaging inflammation. An in vivo inflammation model was prepared by intramuscular (i.m.) injection of Freund's complete adjuvant to the thigh of five rabbits. Serial imaging of the animals was done with 99Tcm-dextran for 2 weeks following the onset of inflammation. The radiotracer was seen to concentrate selectively in the lesion, with a mean target-to-nontarget ratio of 3.01 (range 2.74-3.15) from day 4 onwards. The area of scintiscan abnormality increased with time and was more marked than the physical appearance of the lesion. In two rabbits, the tracer was also seen to localize at the site of injection of the anaesthesia in the normal thigh. The best time to image was found to be 2-4 h after 99Tcm-dextran administration. It would appear that abnormally increased capillary permeability in the inflamed area results in enhanced uptake of the macromolecule. We conclude that 99Tcm-dextran is a potential inflammation-seeking radiotracer, though it needs to be evaluated further. PMID- 8719989 TI - Delayed gastric emptying does not predispose to Helicobacter pylori infection in non-ulcer dyspepsia patients. AB - Non-ulcer dyspepsia is a common disorder in clinical practice. The pathogenesis and predisposing factors that lead to the development of Helicobacter pylori infection are still unclear. The aim of the present study was to evaluate the prevalence of H. pylori infection in non-ulcer dyspepsia patients with delayed gastric emptying and those with normal gastric emptying, and to determine if delayed gastric emptying predisposes to H. pylori infection. A total of 70 patients (29 males, 41 females) aged 18-59 years (mean +/- S.D. 40.5 +/- 11.2 years) took part in the study. A solid-phase scintigraphic gastric emptying study and 14C urea breath test were performed on each patient. There was no statistically significant difference in age between those patients with and without delayed gastric emptying (40.8 +/- 11.9 vs 40.4 +/- 10.5 years), or between those with and without H. pylori infection (40.5 +/- 12.6 vs 40.5 +/- 9.8 years). Of the 70 patients, 45 (64.3%) had delayed gastric emptying and 25 (35.7%) had normal gastric emptying. The overall incidence of H. pylori infection was 58.6%. The incidence of H. pylori infection among patients with delayed gastric emptying was 60% (27/45 patients) and among patients with normal gastric emptying 56% (14/25) patients). The incidence of H. pylori infection in the two groups was not significantly different. The incidence of H. pylori infection in the non-ulcer dyspepsia patients in this study was similar to that of age-matched asymptomatic volunteers in Taiwan. In conclusion, based on the results of this study, delayed gastric emptying does not lead to a predisposition to H. pylori infection in non-ulcer dyspepsia patients. PMID- 8719990 TI - Morphine-augmented hepatobiliary scintigraphy: a meta-analysis. AB - The aim of this study was to compare the sensitivity and specificity of morphine augmented hepatobiliary scintigraphy (MA-HBS) with that of conventional hepatobiliary scintigraphy (C-HBS) for acute cholecystitis. The results of most MA-HBS studies cannot be compared with C-HBS estimates, since articles describing C-HBS often include non-candidates for MA-HBS. However, using meta-analytic techniques to combine data from eligible studies (4 for C-HBS and 5 for MA-HBS), the specificity of MA-HBS (0.84; 95% CI = 0.75-0.94) was significantly greater (P = 0.008) than that of C-HBS (0.68; 95% CI = 0.61-0.75); there were no differences in sensitivity (MA-HBS; 0.96, 95% CI = 0.92-0.99; C-HBS: 0.97, 95% CI = 0.97 0.99). PMID- 8719992 TI - The case for conservative management of infected prosthetic grafts. PMID- 8719991 TI - An overview of the treatment of infected prosthetic vascular grafts. PMID- 8719993 TI - The case against the conservative nonresectional management of infected prosthetic grafts. AB - The perception that there is an expanding role for conservative management of patients with aortic graft infection is unfounded. There is,in fact, a striking paucity of convincing data indicating that outcome following nonresectional therapy for aortic prosthetic graft infection is equivalent to modern-day results utilizing extra-anatomic bypass and graft excision. Drainage with localized antibiotic irrigation and biologic coverage may be attempted in unusual circumstances such as the unfortunate patient with an infected thoracoabdominal aortic graft, in whom graft excision is not feasible. A truly noteworthy development in the treatment of aortic graft infection over the past decade has been the remarkable improvement in results utilizing remote bypass and standard excisional therapy with perioperative mortality and amputation rates less than 10%. In our opinion this approach remains the best and safest option. PMID- 8719994 TI - Management of the short gut syndrome. PMID- 8719995 TI - The management of perianal diseases. PMID- 8719996 TI - The clinical use of gastrointestinal hormones for alimentary tract disease. PMID- 8719998 TI - The management of intrahepatic stones. PMID- 8719997 TI - Endovascular stents and stented grafts for the treatment of aneurysms and other arterial lesions. AB - The combination of prosthetic graft and intravascular stent technologies will probably become an important part of the treatment for aneurysmal and occlusive disease as well as for traumatic vascular injuries. This technology potentially permits reduced operative morbidity and mortality rates as well as decreased intraoperative blood loss, cost, and hospital stay, with ultimately improved patient care. Once additional experience with this important new technique has been obtained, randomized, prospective trials comparing standard therapy to endovascular grafting procedures will be needed to substantiate this form of therapy for the treatment of various arterial lesions. PMID- 8719999 TI - Colonic volvulus. AB - Volvulus has multifactoral causes. It occurs worldwide, and is a common cause of large bowel obstruction. Current knowledge suggests that primary prevention is not possible, so better management is needed if the number of deaths are to be reduced. Most cases of sigmoid volvulus can be nonoperatively decompressed. However, operation is required to remove gangrenous bowel and stop recurrence. Treatment needs to be individualized as appropriate for medically compromised patients. Sound clinical judgment still has an important role in the diagnosis and management of this disorder. PMID- 8720000 TI - Laparoscopic colon resection for cancer--a favorable view. PMID- 8720001 TI - Laparoscopic colon resection for cancer--an unfavorable view. PMID- 8720002 TI - Current uses and abuses of total parenteral nutrition. AB - Total parenteral nutrition remains a vital medical intervention, and in our institution it is considered as basic as intravenous fluids, antibiotics, and blood transfusions. As is true of most treatments, there are specific indications as well as associated risks and costs which mandate justification of its use. It is clear that the indications for TPN are diminishing as basic science and clinical studies continue to find increased benefits associated with enteral feeding, and as techniques for initiating enteral nutrition improve. Because of unproven benefits and/or increased complication rates, TPN has lost favor in the treatment of conditions that were previously thought to require parenteral alimentation, such as acute pancreatitis, pediatric and adult burns, critical care, and preoperative use in patients with mild or moderate malnutrition. Despite diminishing indications, TPN continues to generate excitement in some areas as its immunological effects become better defined. The use of TPN in patients with cancer before certain therapies, as well as in the transplant population, remains hopeful. New uses of TPN will result from a better understanding of the cellular and molecular effects of parenteral feeding. In the future, TPN may well be used as a pharmacologic agent rather than as nutritional intervention. PMID- 8720003 TI - Thrombolytic therapy in peripheral vascular disease. PMID- 8720004 TI - Current management of Meckel's diverticulum. PMID- 8720005 TI - Pelvic exenteration for recurrent rectal cancer. AB - Recurrent rectal carcinoma presents a formidable problem for patient and surgeon. Isolated recurrences of rectal carcinoma have been reported from 7% to 33% with a median of 14%. Increasing recurrence is associated with increasing Dukes's stage. The reason for high recurrences is probably related to the limited anatomic margins that can be obtained in the pelvis during primary resections. Patients who have recurrence after a low-anterior resection are more likely to present with nonfixed, surgically correctable lesions versus recurrences after abdominoperineal resection. The most common symptom related to pelvic recurrence is pain, which may be perineal or radiate to the lower extremities. The 5-year survival rate among unresected patients with locoregional recurrences is 4%. These patients are often in extreme pain with lower extremity swelling and perineal lesions. Although many patients will be palliated by radiation, few will experience long-term relief (6 to 8 months). A thorough physical examination should include rectal and pelvic exams to evaluate tumor extension and fixation. Computed tomographic studies are helpful when taken serially to evaluate pelvic recurrence or liver metastases. Fineneedle biopsies may also be done under CT guidance. Additional mandatory films include plain chest roentgenograph, lumbosacral spine films, and bone scan to rule out sacral involvement, which would preclude sacral resection. Magnetic resonance imaging has recently been shown to be effective in evaluating pelvic side wall involvement and metastatic lymphadenopathy. Although extensive involvement would preclude aggressive resection, in one series, 50% of patients were amenable to resection. Pelvic exenteration should include the tumor mass, along with any involved organs and their lymphatic drainage, with a 2 cm margin. Complications are increased in patients who have undergone radiation, who have undergone procedures that include urinary diversions, and who have recurrent disease. Cure rates of 30% to 50% have been reported using pelvic exenteration for rectal cancer. Recurrent disease presents a significant problem in that normal anatomic planes have been disrupted. In one series, rectal recurrences treated with pelvic exenteration had a 66% recurrence rate. In addition, there is often a posterior component to the recurrence. Although the complication rate is high, the only chance for cure in these patients would be an abdominosacral resection. There appears to be a select group of patients with recurrent locoregional disease, who benefit from sacral resection with a 20% to 30%, 5-year survival rate. PMID- 8720006 TI - Vascular resection and intraoperative radiation therapy during pancreaticoduodenectomy: rationale and technique. PMID- 8720007 TI - Tamoxifen therapy--long-term results and complications. PMID- 8720008 TI - The current management of common bile duct stones. AB - Choledocholithiasis is usually proven by cholangiogram during cholecystectomy, and today nearly all cholecystectomies are performed laparoscopically. Patients expect to be discharged from the hospital within 24 hours after laparoscopic surgery and return to work in 1 week. Surgeons must develop advanced laparoscopic skills to allow choledocholithiasis to be managed laparoscopically with one procedure and one anesthetic. Interventional radiologists and gastroenterologists can frequently detect and manage common duct stones that cause severe cholangitis and pancreatitis and that occur in patients too debilitated for general anesthesia. In the majority of patients, common duct stones demonstrated at laparoscopic cholecystectomy may be removed using laparoscopic transcystic techniques. Whether complicated CBD stones are optimally managed by laparoscopic choledochotomy, conversion to open common duct exploration, or left for postoperative ERC/ES is controversial and depends on the expertise of the surgeon, gastroenterologist, and wishes of the patient. Although the authors propose one algorithm for the current management of CBD stones, the superiority of the laparoscopic treatment for CBD stones is not yet proven. Multi institutional, randomized trials comparing laparoscopic, endoscopic, and combined techniques for ductal clearance will be necessary to establish the optimal therapy of choledocholithiasis in the 1990s. PMID- 8720009 TI - Endoscopic vagotomy for duodenal ulcer disease: procedures and appraisal. PMID- 8720010 TI - Patient compliance for psychotropic medications: a group model for an expanding psychiatric inpatient unit. PMID- 8720011 TI - Preparing for prescriptive practice: Advanced practice psychiatric nursing and psychopharmacotherapy. PMID- 8720012 TI - Living with depression: family members' experiences and treatment needs. PMID- 8720013 TI - Out of control! the most effective way to help the binge-eating patient. AB - This article assessed the effectiveness of different treatments for bulimia nervosa by screening 400 studies published from 1987 through July 1933. Nineteen independent studies with a total of 1,015 subjects with 11 treatment types, and 316 subjects in 11 control groups fulfilled these criteria. Therapy outcome across studies was assessed meta-analytically, producing effect sizes that represent pretreatment to posttreatment changes in bulimic symptoms. PMID- 8720014 TI - The impact of the evolution of biological psychiatry on psychiatric nursing. PMID- 8720015 TI - Format-independent data collection forms. AB - Good form design is generally acknowledged to be a key element of clinical trials and follow-up studies. Although some data are collected directly from automated equipment, most measurements, examination findings, and interview responses require at least one manual transcription, either onto paper forms or directly into a computer. Regardless of the transcription method adopted, there is broad agreement that the quality of study data depends strongly on the format and layout of the data collection instruments. The task of form design and production is complicated by two facts: first, a series of different forms is needed even when the period of follow-up is short; and second, form design is an ongoing process that starts before the first person is enrolled and continues over the course of the trial. This paper (1) presents a concept (format independence) aimed at simplifying the design and revision of data collection forms; (2) explains how a feature (styles) present in most word processing software can be used to implement the concept; (3) discusses the advantages and disadvantages of a format-independent approach as compared to more traditional form development tools; and (4) describes the environment that prompted this approach to form development. PMID- 8720016 TI - The clinical-economic trial: promise, problems, and challenges. AB - The clinical-economic trial is a study design that is appearing with greater frequency in medical and public health literature. Some experienced investigators view these trials with skepticism; to policy makers they represent a promising step in the control of rising health care costs. The success of clinical-economic trials in meeting the important goal of more rational and efficient use of health care resources will depend on the strengths and limitations of the research method. As part of a report to the Office of Technology Assessment of the U.S. Congress on new health care assessment techniques, we describe the reasons why economic data collection and analysis are being considered in clinical trials, identify and discuss various designs and methods for gathering economic trial data, and evaluate the strengths and limitations of different methods for providing sound data for decision making on appropriate use of health care interventions. Because of the potential significance and increasing visibility of such research, experts in research methods should give more attention to methodological research for clinical-economic trials. Future efforts should be directed at comparing different techniques for collecting data, examining the incremental value of precision in economic measurements and ensuring appropriate interpretation of data from clinical-economic trials. PMID- 8720017 TI - Sample size calculation for complex clinical trials with survival endpoints. AB - Sample size estimation is important in planning clinical trials. The purpose of this paper is to describe features and use of SIZE, a comprehensive computer program for calculating sample size, power, and duration of study in clinical trials with time-dependent rates of event, crossover, and loss to follow-up. SIZE covers a wide range of complexities commonly occurring in clinical trials, such as nonproportional hazards, lag in treatment effect, and uncertainties in treatment benefit. The use of SIZE is illustrated by several hypothetical examples as well as applications to real study designs, each featuring a statistical issue. PMID- 8720018 TI - The data and safety monitoring board and acquired immune deficiency syndrome (AIDS) clinical trials. AB - The urgency of the Acquired immune deficiency syndrome (AIDS) epidemic has mandated that multiple therapeutic approaches be developed and that these approaches be evaluated through clinical trials. To oversee these trials, the National Institute of Allergy and Infectious Diseases (NIAID) has created three large clinical trial programs monitored by a Data and Safety Monitoring Board (DSMB). For each clinical trial, this Board uses a standardized approach employing contemporary biostatistical, medical, and ethical principles. The DSMB is responsible for reviewing interim data on clinical trial performance, treatment safety and efficacy, and overall study progress. If interim results provide convincing evidence of either excessive adverse effects or significant treatment benefit, the DSMB may recommend early termination of the trial to the NIAID and the study investigators. The responsibility, organization, and operating procedures of this DSMB are presented and illustrated through three clinical trials sponsored by NIAID and monitored by the Board. The rationale and operational model for the DSMB may be a useful example for the development of similar review processes in other HIV clinical trial settings. PMID- 8720019 TI - Response rates to six recruitment mailing formats and two messages about a nutrition program for women 50-79 years old. AB - Recruiting participants for large prevention trials is time consuming and costly. In order to test various recruitment techniques, we conducted two studies of response rates to recruitment mailings for the Women's Health Trial. The potential participants, 50- to 79-year-old women, were requested to return an enclosed postcard to learn more about the trial. In the first study, we sent at random either a short or a long message to a group of University of Miami personnel (N = 862) and a Dade County cohort (N = 2964). More university women responded to the short message than to the long message (22.4% vs 16.4%, p = 0.024). Similarly, more of the Dade County cohort replied to the short message than to the long one (12.1% vs 9.6%, p = 0.027). The long message listed details of the intervention (e.g., modifying recipes) that some women may have used to decide they were not interested in participating. In the second study, we examined response rates to two different ways of addressing the mailing, i.e., handwritten envelopes and machine-printed labels; we also evaluated three methods for delivering the short message: (1) formal invitation, (2) business letter with an inside name and address of the recipient, and (3) business letter without the recipient's name and address. Response rates were similar between the methods of addressing envelopes and among the three vehicles for the message, suggesting that the least costly method of mailing should be used. PMID- 8720020 TI - Response to S.J. Day letter. PMID- 8720021 TI - Sample size calculations for repeated measures experiments. PMID- 8720022 TI - Medical Research data. PMID- 8720023 TI - Labor force planning issues for allied health in Australia. AB - The aim of this paper is to discuss labor force planning issues for allied health professionals in Australia. Health system reform and changes in the demand for health labor, combined with key characteristics of the professions, will have a profound influence on future needs for career development of allied health professionals. Key issues include the increasing need for allied health professionals to undertake business management and public health training, the growing trend of multiskilling versus specialization, and the need for the professions to diversify their skill base to ensure a range of career options in a changing health care system. The challenge for allied health professions is to improve tools of analysis in relation to labor force planning and to systematically investigate various factors influencing labor force supply and demand, on both a short-term and long-term basis. PMID- 8720025 TI - Index of graduate theses and projects in allied health. PMID- 8720024 TI - Australian physiotherapists' and occupational therapists' views on professional practice. AB - The extent to which physiotherapists and occupational therapists endorse a humanistic, holistic approach to health care was addressed in a survey of attitudes of practicing occupational therapists (N = 378) and physiotherapists (N = 558). A 22-item questionnaire including aspects of therapist's knowledge and techniques, holistic approach, client characteristics, client/therapist relationship, and approaches to professional practice was employed. The findings indicate that the professional practices reflecting the new public health model prevailed. This model emphasizes client/therapist interaction, the client's own ability to change and improve, and holistic attitudes towards health care, along with the therapist's knowledge and skills. However, comparative analyses of the two therapy groups reveal that although both groups support a humanistic approach to client care, occupational therapists promote the client's resources and coping skills to a greater extent, place more emphasis on the client in terms of the client's capacity for recovery and coping abilities, were more involved with the social, personal, and psychological problems of the client, and place less emphasis on treatment methods and techniques than physiotherapists. Demographic variables including gender, age, and workplace setting (public/private) were found to be significantly related to views of professional practice. Recommendations for the training of therapists and the implications for practitioners are made and suggestions for further research are given. PMID- 8720026 TI - Potential patterns--conceptual and practical issues in interdisciplinary education. AB - Interdisciplinary education is, again, a timely topic. It has been seen as an important strategy in recent reports and commissions dealing with the future of the health care system and its personnel needs. The present article reviews conceptual and practical issues in the design, development, and implementation of interdisciplinary education in allied health. A foundation for planning involves the integration of an interdisciplinary practice model with the usual model of psychomotor skills, cognitive skills, and knowledge domain for a particular discipline. Planning issues are addressed in a series of six questions for allied health administrators and faculty to consider. PMID- 8720027 TI - Empirically derived subtypes of pervasive developmental disorders: a cluster analytic study. AB - A cluster analytic study was conducted to empirically derive behaviorally homogeneous subtypes of pervasive developmental disorders (PDD). Subjects were clustered based on a broad range of behavioral symptoms which characterize autism. Behavioral variables were measured using several of the standardized psychometric instruments most commonly employed in assessing autistic individuals. The cluster solution indicated the presence of four distinct groups. Validity checks generally confirmed significant between-group differences on independent measures of social, language, and stereotyped behaviors. In addition, the four-group cluster solution was compared to previously developed typological systems of PDD (i.e., subcategories based on IQ early onset, styles of social interaction, and DSM-III-R diagnosis). Results generally supported both the behavioral homogeneity of the four subgroups and also several important between group differences. The potential utility of using cluster analyses to explore subtypes of PDD is discussed. PMID- 8720028 TI - Social initiations by autistic children to adults and other children. AB - Social initiations made by autistic and verbal-matched retarded children were recorded in two naturalistic situations. Frequencies of initiation to adults did not differ between groups, but the retarded children initiated much more frequently to peers. Most interactions for both groups were positive, but the autistic children engaged in more ritualized, and the retarded children more playful, initiations. The autistic children monitored the social environment more when forced into proximity with peers, whereas the retarded children initiated more in the unstructured situation. Autistic initiation to peers was unrelated to severity of autism, but was related to cognitive skills, including vocabulary and comprehension of affect, whereas retarded children's initiations were unrelated to cognitive level. Results are discussed in terms of the differences between adults and children as social stimuli, prerequisite skills for initiation to peers, and the relationship between social cognition and social behavior. It is suggested that autistic and retarded children differ in the quantity of their initiations to peers, and the quality of their initiations to adults, and that initiations to peers may be a particularly useful index of social development in autistic children. Results confirm the need of autistic children for highly structured social environments, and suggest an important role for the remediation of specific cognitive skills such as comprehension of others' affects. PMID- 8720029 TI - Failure to confirm the word-retrieval problem hypothesis in facilitated communication. AB - Two hypotheses were raised and empirically tested to account for the failure of previous controlled validation studies to find evidence of literacy in nonspeaking persons with autism using facilitated communication: (a) The naming tasks used in other studies have triggered specific "word retrieval" problems, or anomia, and (b) a perceptual problem, visual agnosia, prevents subjects from recognizing objects without touching them. Three nonspeaking autistic children who had used facilitation for at least 2 years were evaluated with four experimentally controlled tasks, over a period of 5 months. In descriptive and object handling tasks, and in a traditional picture identification task, subjects failed to type correct answers when facilitators were blind; one subject, however, occasionally engaged in signing and vocalizations that were context appropriate. Results reflected a generalized language deficit, rather than isolated word-finding or perceptual difficulties, and were consistent with many previous studies revealing facilitator cuing. Questions are raised about inconsistencies in pseudo-correct scores, a measure of facilitator influence, reported here and in previous research. PMID- 8720031 TI - Adolescent schizophrenia: a methodologic review of the current neuroimaging and neuropsychologic literature. AB - This paper reviews all relevant articles that reported structural neuroimaging or neuropsychological data in adolescent patients with schizophrenia. These papers were subsequently examined from a methodological perspective. Few papers have been written that have examined whether adolescent schizophrenia is associated with structural neuroimaging abnormalities or cognitive dysfunction. In these studies, major methodologic issues exist. Therefore, at present, firm conclusions cannot be made regarding the presence or absence of neuropsychologic dysfunction or structural neuroimaging abnormalities in this population. Attention to certain methodologic issues may improve future studies of this topic. PMID- 8720030 TI - The effect of clozapine on self-injurious behavior. AB - Traditional neuroleptic drugs like thioridazine and haloperidol have not proven to be systematically effective with the treatment of self-injurious behavior (SIB). These drugs may be ineffective because they primarily block D2 dopamine receptors. Based on research with humans and other animals, it appears that another dopamine receptor, D1, may be responsible for mediating some SIB. Clozapine, a neuroleptic recently introduced in the United States, has proven effective in treatment of refractory cases of schizophrenia and is known to have an affinity for blocking D1 receptors. The drug was used to complete a 93-week double-blind crossover trial with a client displaying chronic SIB. Though clozapine is known to affect other neurotransmitter systems, the successful treatment of the participant is consistent with the D1 hypothesis of self injurious behavior and suggests the possibility that clozapine could be an effective pharmacological intervention for some cases of SIB. PMID- 8720033 TI - Brief report: the association between autism and fragile X syndrome: a case report. PMID- 8720032 TI - Brief report: circadian melatonin, thyroid-stimulating hormone, prolactin, and cortisol levels in serum of young adults with autism. AB - An abnormal circadian pattern of melatonin was found in a group of young adults with an extreme autism syndrome. Although not out of phase, the serum melatonin levels differed from normal in amplitude and mesor. Marginal changes in diurnal rhythms of serum TSH and possibly prolactin were also recorded. Subjects with seizures tended to have an abnormal pattern of melatonin correlated with EEG changes. In others, a parallel was evidenced between thyroid function and impairment in verbal communication. There appears to be a tendency for various types of neuroendocrinological abnormalities in autistics, and melatonin, as well as possibly TSH and perhaps prolactin, could serve as biochemical variables of the biological parameters of the disease. PMID- 8720034 TI - The use of buspirone with aggressive behavior. PMID- 8720035 TI - Experimental infection of SPF piglets with porcine reproductive and respiratory syndrome (PRRS) viruses isolated from two farms. AB - Porcine reproductive and respiratory syndrome (PRRS) viruses were isolated from pig samples obtained from two farms characterized by an increased number of stillbirth and high mortality in new-born piglets (farm A), and respiratory distress with high mortality in weaning and growing pigs (farm B) in 1993, respectively. When primary specific pathogen-free piglets, 5-day-old or 13-day old, were experimentally inoculated with the isolates, they showed clinical signs of depress, anorexia, pyrexia, diarrhea, sitting posture and periocular edema. Rate of the weight gain was reduced in the inoculated piglets compared with the non-inoculated pig. There were no apparent differences in clinical signs between the piglets inoculated with the virus samples derived from farms A and B. Microscopically, the most prominent changes observed in experimentally inoculated piglets were interstitial pneumonia, nonpurulent myocarditis and catarrhal lymphnoditis post inoculation day (PID) 28. Viruses were recovered from tissues collected from the inoculated piglets on PID 7 or 28. Furthermore, the viruses were continuously recovered from the sera from PID 7 to PID 28. Antibodies measured by indirect immunofluorescence assay to PRRS virus were first detected in sera on PID 14, and the antibody titer rose to 1:1280 on PID 28. PMID- 8720036 TI - Renal responses to atrial natriuretic peptide (ANP) in rats with non-oliguric acute renal failure induced by cisplatin. AB - This study was designed to compare the renal effects of atrial (A-type) natriuretic peptide (ANP) on control (saline-injected) rats and rats with non oliguric acute renal failure induced by cisplatin. The results obtained here are summarized as follows: (1) In the metabolic cage study, cisplatin-treated rats showed increases in blood urea nitrogen and serum creatinine while creatinine clearance decreased to the lowest levels on day 4. A transient increase in urinary protein was observed at day 4. (2) ANP infusion significantly increased urine flow rate (UFR), creatinine clearance (CCr), fractional excretion rates of sodium (FENa) and chloride (FECl), and urinary phosphorus and magnesium (Mg) excretions in a dose-dependent manner without affecting renal plasma flow and fractional excretion rates of potassium and urea in cisplatin-treated rats. (3) Renal effects of ANP on UFR, CCr, FENa, FECl and excretion of Mg were more pronounced in cisplatin-treated rats compared to control rats although markedly blunted responses to ANP have been reported in nephrotic patients and nephrotic animals induced by adriamycin and aminonucleoside. (4) Histological examination showed extensive necrosis of the S3 segment of the proximal tubule located in the outer stripe of the outer medulla with minimal glomerular abnormalities in the kidney of cisplatin-treated rats. In conclusion, the main mechanism of the increased renal responses to ANP is considered to be due to an increased delivery of sodium, fluid and ANP itself to the inner medullary collecting duct which is the major renal site of action of ANP under the condition of acute proximal tubular necrosis by cisplatin. PMID- 8720037 TI - Resistibility to Theileria sergenti infection in Holstein and Japanese Black cattle. AB - To clarify the resistibility against Theileria sergenti infection in Holstein and Japanese Black cattle under controlled conditions in artificial environmental chambers, those breeds of calves were infected with T. sergenti by infestation with the same number of infected ticks. No apparent differences between the two breeds were observed in the feeding numbers and feeding periods of ticks infested. The level and duration of parasitemia were clearly less and shorter in Japanese Black than those in Holstein calves. With progress of parasitemia, packed cell volume (PCV) and erythrocyte number decreased in both breeds. However, the minimum PCV and erythrocyte number recorded in Japanese Black were higher than those in Holstein calves. These results suggest that the Japanese Black shows solider resistance to T. sergenti infection than the Holstein breed does under experimentally controlled condition without external factors. PMID- 8720038 TI - Rapid inhalation induction of anesthesia by halothane, enflurane, isoflurane and sevoflurane and their cardiopulmonary effects in dogs. AB - The rapid inhalation induction of anesthesia (RII) by mask inhalation of halothane, enflurane, isoflurane and sevoflurane at an equianesthetic concentration (2.5 MAC) was evaluated in 24 beagle dogs. The differences in movements, induction and intubation time between anesthetics were mainly associated with the differences in each blood/gas solubility. The most rapid and smoothest induction was observed by sevoflurane inhalation (209.0 +/- 44.2 sec), followed by isoflurane inhalation (285.8 +/- 34.1 sec). Halothane inhalation took the longest induction time (790.3 +/- 75.7 sec). Movements during RII were minimal in sevoflurane group comparing to the other groups. Heart rate, cardiac output and rate pressure product significantly increased after the beginning of inhalation in all the dogs except for those of halothane group. These changes exceeded the physiological level just after the beginning of inhalation, however, rapidly reversed to the maintenance level (1.5 MAC) approximately 10 min after intubation. Consequently, sevoflurane seemed to be the best inhalational anesthetic for RII in dogs without significant problems in respiratory and/or cardiac functions. Isoflurane also induced rapid induction with some degree of the movements. PMID- 8720039 TI - Attachment, outgrowth, invasion and formation of the egg cylinder in mouse half embryos in vitro. AB - To investigate the effect of the decrease in the number of embryonic cells on the development and morphogenesis of mouse embryos at the peri-implantation stage, half embryos which developed from bisected morulae were morphologically classified into eu-blastocysts (eu-blasts), pseudo-blastocysts (pseudo-blasts), trophectodermal vesicles (TVs) and non-integrated forms (NIFs) and then cultured on plastic substratum, uterine-epithelial-cell monolayer and type I collagen gel. When half embryos were cultured on plastic substratum and cell monolayer, the rates of attachment and trophoblast outgrowth in the eu- and pseudo-blasts were not significantly different from those of the controls. The TVs and NIFs showed significantly lower rates of outgrowth than the controls (P < 0.01). When half embryos were cultured on type I collagen gel, no significant difference was observed in the rate of primitive endoderm formation between the eu-blasts and controls after 36 hr of culture. In the eu-blasts, however, the developmental rate to the egg cylinder at stage 8 was significantly lower than in the controls after 72 hr (P < 0.05). The pseudo-blasts revealed significantly lower rates of endoderm formation and development to the egg cylinder than the controls after 36 and 72 hr, respectively (P < 0.05). In the TVs and NIFs, the rates of outgrowth were significantly lower than those of the controls (P < 0.05) and no egg cylinder was observed. The invasion of type I collagen gel by the cytoplasmic protrusions of trophoblast cells was observed regardless of the type and developmental stage of the embryos. The results show that a decrease in the number of embryonic cells affects the formation of the primitive endoderm and the development to the egg cylinder in vitro. PMID- 8720041 TI - Validity of retrograde corpora lutea of pregnancy as an index of past gestation in Sika deer, Cervus nippon. AB - Corpora lutea of the sika deer (Cervus nippon, Cervidae) were histologically examined in order to elucidate their retrograding processes after parturition. They showed a remarkable regression within one month as luteal cells decreased in number and, by nine months after parturition, turned into masses of blood vessels containing many arterioles with a diameter of 30 microns or less. In retrograde corpora lutea (RCL), older than 1.5 years after parturition, arterioles of these dimensions decreased markedly, while those measuring 50 microns or more became dominant. These findings indicate that an examination of RCL can distinguish females less than nine months after parturition from those more than 1.5 years after parturition. Based on the findings of RCL from the ovaries of a 10.5-year old female, it is suggested that RCL remain in the ovaries for at least 8.5 years after parturition. It was, however, difficult to distinguish RCL of pregnancy from retrograde accessory corpora lutea (RACL). In order to arrive at a complete count of RCL, the ovaries should be examined at thicknesses of 0.5 mm. To check the presence/absence of RCL of pregnancy within nine months or less after parturition, sectioning at thicknesses of 2 mm is sufficient. PMID- 8720040 TI - Antigenic and genetic analyses of the hemagglutinin of influenza viruses isolated from pigs in 1993. AB - Three strains of influenza A virus (H3N2) were isolated from pigs in Hokkaido, Japan in 1993. The hemagglutinin (HA) antigen of the three isolates was related to that of recent H3N2 viruses of human origin. The reactivity patterns of two of the isolates (A/sw/Obihiro/1/93 and A/sw/Obihiro/2/93) with monoclonal antibodies to the hemagglutinin of A/Bangkok/1/79 strain were similar to that of the human H3N2 strain isolated in Hokkaido in 1988, while that of the other one (A/sw/Obihiro/3/93) was similar to human H3N2 strains of 1993. In the phylogenetic analysis based on the nucleotide sequences of the HA1 regions, the HA genes of the two isolates were shown to be closely related to those of human H3N2 viruses isolated between 1986 and 1988. The remaining one isolate was shown to be closely related to those of current human H3N2 viruses. We have also found serological evidence that the A/sw/Obihiro/1/93 virus is circulating extensively in Obihiro swine. It is clear from these findings that pigs were infected with the recent H3N2 influenza virus during the human epidemic and that the virus has been maintained in pigs for at least five years. PMID- 8720042 TI - Ultrastructure of cardiac myocyte in the Asian elephant (Elephas maximus). AB - Cardiac myocytes of an Asian elephant (Elephas maximus) were observed by transmission electron microscopy. Typical ultrastructural features of cardiac myocytes are exhibited in the musculature of both the left and right atria, and left ventricle of the heart. Myofibrils, mitochondria, T-system and sarcoplasmic reticulum are well-developed within the cytoplasm. Many mitochondria are characteristically concentrated is some myocytes. Cardiac musculature is also distributed in the root of the caudal vena cava. Many atrial granules are detected not only in atrial myocytes, but also in the myocytes of the caudal vena cava. Atrial natriuretic polypeptide may be secreted from the caval venous wall in the elephant. PMID- 8720043 TI - Changes in the behavioral parameters following the lipopolysaccharide administration in goats. AB - The present study was aimed at the establishment of an experimental model for the numerical assessment of sick animal behavior. Four goats were given bolus injections of 200 ng/kg of lipopolysaccharide (LPS) or vehicle (0 hr) under non restrained conditions, and observed for behavioral changes and clinical symptoms during the period between -1 and 10 hr. The apparent clinical symptoms of miosis and shivering were observed during the period from 39.5 +/- 3.1 to 296.5 +/- 9.9 min and from 46.0 +/- 2.3 to 251.0 +/- 15.5 min after the LPS administration, respectively. As to the general behaviors, the total length for standing and sternum lying during the period from 0 to 5 hr after LPS administration showed no change, however, that for feeding and rumination, and the cumulative number of grooming episode were significantly reduced as compared to the control period. On the other hand, the cumulative numbers of urination, defecation and yawning showed a tendency to increase but not significantly. These results suggest that stereotyped behavioral responses, which are typically seen in acute phase of sickness, can be transiently induced in goats by treating them with LPS. PMID- 8720044 TI - Ventricular wall thickness and blood pressure values in normal cynomolgus monkeys. AB - In order to clarify the relationship between heart morphology and blood pressure, I examined the mutual correlation between the ventricular wall thicknesses of heart and body weight, heart weight, and arterial blood pressure in 15 normal male cynomolgus monkeys. The results obtained were as follows; (1) The mean and standard deviation of the arterial blood pressure was 68 +/- 8 mmHg at diastolic time and 124 +/- 12 mmHg at systolic time, respectively, and that of pulse rate was 227 +/- 17 beats per min under non-anesthesia. In addition, the mean and standard deviation of the arterial blood pressure was 55 +/- 11 mmHg at diastolic time and 102 +/- 18 mmHg at systolic time, respectively, and that of pulse rate was 197 +/- 17 beats per min under general anesthesia. (2) The mean and standard deviation of the left ventricular wall thickness was 6.6 +/- 1.1 mm at anterior, 5.3 +/- 1.0 mm at posterior, 5.9 +/- 0.9 mm at lateral, 5.4 +/- 1.2 mm at the interventricular septum, and that of the right ventricular wall thickness was 2.4 +/- 0.5 mm at anterior. (3) The mean and standard deviation of body weight was 4.1 +/- 0.7 kg, and that of heart weight was 16.5 +/- 3.1 g. (4) The heart weight and interventricular septum thickness correlated significantly to body weight, respectively. (5) There was no significant correlation between arterial blood pressure and the ventricular wall thickness. Further study should be necessary in more normal monkeys, together with abnormal ones with hypertension or cardiac hypertrophy, in order to evaluate the relationship between heart morphology and blood pressure. PMID- 8720045 TI - Toxicity and properties of the extract from Sarcocystis cruzi cysts. AB - The extract from Sarcocystis cruzi cysts in bovine muscle was subcutaneously injected to mice, guinea pigs, chickens, and rabbits to detect its toxicity. Only rabbits showed reactions after administration of the extract at a dose of 25 micrograms. The main clinical signs of the rabbits were depression, reduction in body temperature and intermittent diarrhea and the hematological findings observed were elevation in WBC, RBC, PCV, TP, BUN, AST, AUT and creatinine values and reduction in glucose, K+ and pH of blood. The extract, crude toxin, was a water soluble, acid-alkali stable and thermolabile protein and estimated to be a molecular mass of 15-16 kd. PMID- 8720046 TI - An immunohistochemical study of chick primordial germ cells using the mouse antiserum against chick embryonic gonads. AB - We established a mouse antiserum by immunizing BALB/c mice with sexually undifferentiated 6-day chick embryonic gonads. The antiserum had a higher affinity to the plasma membrane and cytoplasm of primordial germ cells (PGCs) than to the somatic cells in 6-day chick gonads. PGCs at this stage appear to have a higher specificity with more antigenic capacity than gonadal somatic cells; thus, the antiserum should be useful as a cell marker of chick PGCs. Subsequently, an immunohistochemical examination of PGCs settling in the genital ridges of chick embryos was carried out by the use of the antiserum. The reaction was clearly detected in the plasma membrane and cytoplasm of 5-day PGCs. After sexual differentiation, the reaction changed in distribution in females. At 8 days, it was limited to the perinuclear region of PGCs. In males, although the reaction persisted for longer period than in females, only a weak reaction was detectable at 9 days. These results suggest that the occurrence of periodic molecular changes may be different between male and female PGCs. PMID- 8720047 TI - Evaluation of anthelmintic efficacy of doramectin against gastrointestinal nematodes by fecal examination in cattle in Japan. AB - The nematocidal effect of doramectin was assessed by subcutaneous injection of 200 micrograms/kg (single dosage) in cattle with naturally acquired gastrointestinal nematode infection in Japan. This study consisted of two experiments. In experiment 1, animals were randomly divided into a doramectin group (n = 21) and a non-treated control group (n = 21) by fecal egg count. In experiment 2, another group of ivermectin treatment (n = 12) was prepared in addition to doramectin (n = 23) and control (n = 10) groups, by random assignment as in study 1. After doramectin or ivermectin treatment, the egg count/5 g of feces was measured by the sucrose centrifugal flotation method at intervals of 7 days until Day 21 in experiment 1 and at Days 7, 14, 21, 35, 42, 49 and 63 in experiment 2. Coproculture was also carried out using some of the fecal samples. In the 2 doramectin-treated groups, 96.1%-100% egg reduction rates were obtained for Haemonchus, Cooperia, Mecistocirrus, Trichostrongylus, Ostertagia, Bunostomum, Strongyloides and Trichuris from the 7th day until the 49th day after treatment. Thus doramectin was confirmed to be highly effective against those species of adult nematoda. The effect against Mecistocirrus has not been previously determined. The nematocidal effect against Nematodirus was lower (egg reduction rate approximately 50%) than other species. No adverse reactions to treatment were seen in any animal during either study. PMID- 8720048 TI - Distribution of antibodies to influenza A virus in chickens in Japan. AB - A serological surveillance was carried out to detect antibody against influenza A virus in chicken sera. A total of 8,850 field samples were collected from 47 prefectures in Japan. Initially, all the sera were screened by agar gel immunodiffusion and those sera showing positive reaction were investigated for haemagglutination-inhibition (HI) and neuraminidase-inhibition antibodies against influenza viruses. Only 6 samples had antibodies; 4 sera had antibodies against human subtype H1N1 virus; with HI activity against strain A/PR/34; three sera had strong HI activity to strain A/Tottori/4/87, which by haemagglutination test is closely related to A/Yamagata/120/86. The remaining two chicken sera had antibodies against avian subtypes H10N4 and H3N6 viruses respectively. PMID- 8720049 TI - Muscarinic receptor subtypes mediating catecholamine secretion and increase in intracellular Ca2+ concentration in adrenal chromaffin cells of the guinea pig. AB - Muscarinic receptor subtypes mediating catecholamine secretion and increase in the intracellular concentration of Ca2+ ([Ca2+]i) were examined using muscarinic agonists and antagonists in dispersed adrenal chromaffin cells of the guinea pig. All muscarinic agonists (1-1,000 microM) tested caused increases in adrenaline secretion in a dose-dependent manner. Muscarine and methacholine were more effective than bethanechol, oxotremorine and pilocarpine. Muscarine and oxotremorine caused a small increase in adrenaline secretion even in the absence of extracellular Ca2+. Both 4-DAMP (0.1 microM) and pirenzepine (0.1 microM), but not methoctramine (0.1 microM), shifted the dose-response curve for muscarine induced adrenaline secretion to the right. These muscarinic agonists also caused increase in [Ca2+]i in the presence of extracellular Ca2+. Muscarine-induced [Ca2+]i rises were reduced, but not abolished, by removal of extracellular Ca2+. These results suggest that catecholamine secretion induced by muscarinic agonists is mediated through M1, or M1 and M3 muscarinic receptor subtypes in adrenal chromaffin cells of the guinea pig. PMID- 8720051 TI - Activation of complement in quails bearing Rous sarcoma virus-induced tumors. AB - The concentration of serum C3 in quails bearing tumors induced by Rous sarcoma virus (RSV) was elevated in parallel with tumor growth, whereas serum C3 levels in quails inoculated with avian leukosis virus, which lacks transforming activity, showed a pattern similar to that in mock-infected quails. C3 deposition was also observed in almost all tumor cells at the tumor developing stage. These findings obtained in vivo suggest that the cells transformed by RSV activated the C3 on their surfaces. PMID- 8720050 TI - Effects of 6-Hydroxydopamine on the antibody response to the hapten dinitrophenyl in the chicken. AB - Antibody responses to the hapten dinitrophenyl (DNP) and the concentration of catecholamine in chickens received a single injection of 6-Hydroxydopamine (6 OHDA) into the chorioallantoic cavity at the embryonal stage were evaluated. A significantly enhanced anti-DNP IgA response and a markedly decreased level of noradrenaline in the peripheral blood were observed in chickens treated with 400 micrograms of 6-OHDA at 14 days of incubation. These results suggested the immunomodulatory influence of the sympathetic nervous system. PMID- 8720052 TI - Inhibition of mouse hepatitis virus multiplication by phosphorothioate analogues of oligodeoxynucleotides complementary to the leader RNA. AB - Phosphorothioate oligodeoxynucleotides (PS-oligo) complementary to a leader RNA of mouse hepatitis virus (MHV) were more effective inhibitors of MHV multiplication than natural oligodeoxynucleotides (PO-oligo). Sequence-dependent inhibition of viral multiplication was shown at low concentrations (0.001-0.1 microM) of antisense PS-oligo. Phosphorothioate oligodeoxycytidine, PS-(dC)20 and PS-oligo, which has no significant homology to the MHV sequence, showed inhibitory effects on MHV multiplication at concentrations higher than 0.5 microM. These results showed that PS-oligo was more potent than PO-oligo in inhibition of MHV multiplication and that PS-oligo may inhibit MHV multiplication by two different mechanisms, that is, in sequence-dependent and -independent manners. PMID- 8720053 TI - Isolation of Campylobacter spp. from slaughtered cattle and swine on blood-free selective medium. AB - The charcoal-cefazolin-sodium deoxycholate agar (CCDA) was compared with Butzler agar for selectivity of campylobacters from fecal samples. A total of 279 samples of cecal contents from 176 cattle and 103 pigs slaughtered in Saitama were examined. Fifty-five (31.3%) cattle were found to contain campylobacters by the direct plating culture on CCDA plates, compared with 29 (16.5%) on Butzler agar plates. The positive rates of pig samples on CCDA and Butzler agar were very high, being 96 (93.2%) and 86 (83.5%), respectively. After enrichment culture of 107 cattle specimens in Preston, CEM or Bu 10 broth, the isolation rate on CCDA was also higher (p < 0.01) than that on Butzler agar after enrichment in any medium. These results indicate that CCDA medium is easy to handle without blood and, as to this study shows, is superior to Butzler agar for isolation of Campylobacter species from fecal samples. PMID- 8720054 TI - Effects of oil adjuvant on systemic response to Escherichia coli lipopolysaccharide in swine. AB - Intramuscular injection of 0.1 mg/kg of Escherichia coli lipopolysaccharide (LPS) mixed with Freund's complete adjuvant (LPS+FCA) in piglets mitigated the leukopenia and TNF-alpha and cortisol levels in the serum compared with that of LPS suspended in LPS-free saline. The endotoxin level in the serum of the LPS+FCA was remarkably reduced. These results suggest that the addition of oil adjuvant mitigate the systemic toxicity of LPS. PMID- 8720055 TI - Effectiveness of an inactivated goose parvovirus vaccine in Muscovy ducks. AB - With goose parvovirus (GPV) strain IH, an inactivated vaccine was prepared from allantoic fluid of embryonating Muscovy duck eggs inoculated with GPV. The response to vaccination was measured by virus neutralizing antibody titer against GPV. Offsprings from the vaccinated flock were introduced in a farm in which GPV infection had been experienced and examined for resistance to exposure to GPV. The results showed that the intramuscular vaccination to Muscovy ducks at any age stimulated significant virus neutralizing antibody levels, and that more than 90% Muscovy ducklings from the vaccinated parent flock survived after the exposure to GPV. Muscovy ducklings that passively possessed high level virus neutralizing antibodies also could respond to the vaccination and the induced antibodies remained for more than 2 months. PMID- 8720056 TI - A canine case of discoid lupus erythematosus with circulating autoantibody. AB - A nine-year-old Shetland sheepdog was diagnosed as discoid lupus erythematosus by clinical features, histopathologic findings, positive direct immunofluorescence, negative antinuclear antigen test and the absence of multisystemic diseases. The indirect immunofluorescence test of this patient dog with the salt split skin showed the deposition at the bottom of the cleft at basement membrane zone (BMZ). Western immunoblotting revealed the 120 kDa and the 85 kDa proteins targeted by the autoantibody. These proteins did not correspond with the known BMZ component. PMID- 8720057 TI - Compression of spinal cord caused by spinal deformation in a cat. AB - An 8-month-old cat with bilateral hindlimb paresis was admitted. Radiography revealed a curvature of the thoracic spine and a partial deformation of thoracolumbar vertebrae (dorsal extrusion of the cranial and caudal edges). Compression of the spinal cord (T9-L1 and L6-7) was found by myelography. Medicinal treatment did not yield the desired result. Hemilaminectomy and laminectomy were done to relieve pressure on the spinal cord leading to improved gait. It is, however, not clear whether this syndrome was primarily caused by metabolic or genetic factors. PMID- 8720058 TI - The relationship between plasma insulin-like growth factor-I (IGF-I) level and body weight in the horse. AB - The relationship between plasma insulin-like growth factor-I (IGF-I) level and body weight was studied in six breeds of horse. The correlation coefficients between plasma IGF-I and body weight in males and females of 1 year old heavy horses (Percheron and Breton breeds) were 0.62 and 0.12 respectively. The mean plasma IGF-I of males was higher than that of females (p < 0.01). However, the plasma IGF-I levels of heavy horses did not exceed those of light horses (Thoroughbred) or ponies (Shetland and Falabella). These data suggest that IGF-I is at least related to the difference of body weights between sexes in heavy horses. PMID- 8720059 TI - Insulin and insulin-like growth factor-I (IGF-I) stimulate the development of bovine embryos fertilized in vitro. AB - To study the effects of insulin and insulin-like growth factor-I (IGF-I) on early development of bovine embryos, one-cell embryos were cultured in a chemically defined, protein-free medium: modified synthetic oviduct fluid. The addition of 10 micrograms/ml insulin or 20 ng/ml IGF-I increased the percentage of morulae at Day 5 (5 days after insemination). The cell number of blastocysts which developed after 2 days culture of morulae obtained at Day 5 was increased by supplementation with insulin or IGF-I. The combination of insulin and IGF-I had neither additive nor synergistic effect on the embryonic development. These results clearly show that insulin and IGF-I improve the in vitro development of bovine embryos. PMID- 8720060 TI - Effects of human menopausal gonadotrophin (hMG) for superovulation in Japanese Black cattle. AB - Sixty-four Japanese Black parous cows were divided into 5 dosage groups (I, II, III, IV, V) and administered either human menopausal gonadotrophin (hMG) or FSH by intramuscular injection. No significant differences were observed between the hMG-treated groups. The average number of recovered ova/embryos in cows treated with 900 IU was significantly greater (P < 0.05) than in cows treated with FSH. The percentage of high-quality transferable embryos was highest in the 600 IU hMG treated group. Also, FSH-treated cows produced a significantly larger number of unfertilized ova compared to hMG-treated cows. The results suggest that hMG treatment is sufficient for Japanese Black Cattle at a dose of 600 IU for 3 days, and that a 2-day treatment regime is also possible. PMID- 8720061 TI - Establishment and characterization of a cell line (NB-YK) derived from a transplantable rat nephroblastoma. AB - A new cell line, designated NB-YK, was established from a transplantable rat nephroblastoma (NB-Y) which was derived from a spontaneous nephroblastoma in an aged Fischer 344 rat. NB-YK grew in a piling-up and noncohesive pattern on the plastic surface and formed colonies in a soft agar. The main cell type of NB-YK represented morphology of mesenchymal phenotype and most of the cells contained several secretory granules in their cytoplasm. Immunocytochemically the cells were positive for vimentin, cytokeratin, and laminin. Coexpression of vimentin and cytokeratin in the cells was confirmed by the one-dimensional gel electrophoresis and immunoblotting for intermediate filament proteins. NB-YK cells were tumorigenic and produced fibrosarcoma-like tumor when inoculated subcutaneously or intraperitoneally into syngeneic rats and nude mice. NB-YK seems to be a useful model for studying biological properties of nephroblastoma. PMID- 8720062 TI - Clinical application of rapid inhalation induction of anesthesia using isoflurane and sevoflurane with nitrous oxide in dogs. AB - Clinical usefulness of rapid inhalation induction of anesthesia (RII) using 5.0 vol.% isoflurane (GOI) and sevoflurane (GOS) with N2O was evaluated in 124 canine patients. Induction times, loss of various reflexes and movement times were significantly faster in GOI group than in GOS group. Scores for the easiness of intubation and the degree of movements were also significantly less in GOI group than in GOS group. Positive correlation between body weight and degree of movements was observed in both induction groups. Thus, RII using GOI is a practically useful and safe induction modality in small- to medium-sized dogs. PMID- 8720063 TI - Protective effect of the combined vaccine prepared from cell-free-antigen of Actinobacillus pleuropneumoniae serotypes 1, 2 and 5 in pigs. AB - Cell-free-antigens prepared from a concentrated culture supernatant of Actinobacillus pleuropneumoniae (A. pleuropneumoniae) serotypes 1, 2 and 5 were mixed and emulsified with oil adjuvant. The combined vaccine of these 3 serotypes of A. pleuropneumoniae was tested for its ability to confer protection. Pigs immunized with the combined vaccine survived and showed no clinical signs against an intratracheal challenge with A. pleuropneumoniae. In contrast, control pigs inoculated with concentrated culture media emulsified with oil adjuvant developed typical symptoms of pleuropneumonia after challenge inoculation. PMID- 8720065 TI - The chromosome ends of Saccharomyces cerevisiae. AB - Yeast chromosome ends are similar in structure and function to chromosome ends in most, if not all, eukaryotic organisms. There is a G-rich terminal repeat at the ends which is maintained by telomerase. In addition to the classical functions of protecting the end from degradation and end-to-end fusions, and completing replication, yeast telomeres have several interesting properties including: non nucleosomal chromatin structure; transcriptional position effect variegation for genes with adjacent telomeres; nuclear peripheral localization; apparent physical clustering; non-random recombinational interactions. A number of genes have been identified that are involved in modifying one or more of these properties. These include genes involved in general DNA metabolism, chromatin structure and telomere maintenance. Adjacent to the terminal repeat is a mosaic of middle repetitive elements that exhibit a great deal of polymorphism both between individual strains and among different chromosome ends. Much of the sequence redundancy in the yeast genome is found in the sub-telomeric regions (within the last 25 kb of each end). The sub-telomeric regions are generally low in gene density, low in transcription, low in recombination, and they are late replicating. The only element which appears to be shared by all chromosome ends is part of the previously defined X element containing an ARS consensus. Most of the 'core' X elements also contain an Abf1p binding site and a URS1-like element, which may have consequences for the chromatin structure, nuclear architecture and transcription of native telomeres. Possible functions of sub-telomeric repeats include: fillers for increasing chromosome size to some minimum threshold level necessary for chromosome stability; barrier against transcriptional silencing; a suitable region for adaptive amplification of genes; secondary mechanism of telomere maintenance via recombination when telomerase activity is absent. PMID- 8720064 TI - The feline herpesvirus type 1 ICP4 down-regulates feline immunodeficiency virus long terminal repeat (LTR)-directed gene expression via the C/EBP site in the LTR. AB - We investigated effects of feline herpesvirus type 1 (FHV-1) ICP4 on feline immunodeficiency virus (FIV) long terminal repeat (LTR)-directed gene expression by transient transfection assay in Crandell feline kidney cells. We demonstrated that FHV-1 ICP4 significantly stimulates the FIV LTR after introduction of site specific mutation of the C/EBP site in the LTR, and the C/EBP site is sufficient to confer inhibitory effects by FHV-1 ICP4 on a heterologous promoter. These results indicate that FHV-1 ICP4 possesses both ability to transactivate FIV LTR directed gene expression and to down-regulate the FIV LTR via the C/EBP site. PMID- 8720066 TI - An overview of membrane transport proteins in Saccharomyces cerevisiae. AB - All eukaryotic cells contain a wide variety of proteins embedded in the plasma and internal membranes, which ensure transmembrane solute transport. It is now established that a large proportion of these transport proteins can be grouped into families apparently conserved throughout organisms. This article presents the data of an in silicio analysis aimed at establishing a preliminary classification of membrane transport proteins in Saccharomyces cerevisiae. This analysis was conducted at a time when about 65% of all yeast genes were available in public databases. In addition to approximately 60 transport proteins whose function was at least partially known, approximately 100 deduced protein sequences of unknown function display significant sequence similarity to membrane transport proteins characterized in yeast and/or other organisms. While some protein families have been well characterized by classical genetic experimental approaches, others have largely if not totally escaped characterization. The proteins revealed by this in silicio analysis also include a putative K+ channel, proteins similar to aquaporins of plant and animal origin, proteins similar to Na+-solute symporters, a protein very similar to electroneural cation-chloride cotransporters, and a putative Na+-H+ antiporter. A new research area is anticipated: the functional analysis of many transport proteins whose existence was revealed by genome sequencing. PMID- 8720067 TI - Genetically-modified brewing yeasts for the 21st century. Progress to date. AB - Academic studies and traditional breeding of yeasts depend upon their sporulation lifestyle. The strains used have been specially selected to sporulate readily and to mate producing new yeast types. Unfortunately brewing yeast strains do not behave in this way. They sporulate poorly, any spores which are formed are usually non-viable and any haploid strains produced are invariably non-maters. Only in recent years, with the development of recombinant-DNA techniques, has the specific breeding of new brewing yeast strains become widespread. Strains have been produced with the ability to ferment a wider range of carbohydrates, with altered flocculation properties and which produce beers with modified flavours. Many have been tested on the pilot scale and one, an amylolytic brewing yeast, has received approval for commercial use. PMID- 8720068 TI - Processing of pre-ribosomal RNA in Saccharomyces cerevisiae. AB - Post-transcriptional processing of precursor-ribosomal RNA comprises a complex pathway of endonucleolytic cleavages, exonucleolytic digestion and covalent modifications. The general order of the various processing steps is well conserved in eukaryotic cells, but the underlying mechanisms are largely unknown. Recent analysis of pre-rRNA processing, mainly in the yeast Saccharomyces cerevisiae, has significantly improved our understanding of this important cellular activity. Here we will review the data that have led to our current picture of yeast pre-rRNA processing. PMID- 8720069 TI - The initiation of DNA replication in the budding yeast cell division cycle. PMID- 8720070 TI - [PSI] and [URE3] as yeast prions. AB - [URE3] is a non-Mendelian genetic element that mimics recessive mutations in the chromosomal URE2 gene making cells derepressed for nitrogen catabolic enzymes. [PSI] is a non-Mendelian enhancer of readthrough of translational termination similar in its effects to some mutations in the chromosomal SUP35 gene. Three lines of evidence led to the proposal that both [URE3] and [PSI] are prions, infectious proteins analogous to the scrapie agent mediating transmissible spongiform encephalopathies of mammals. 1) Both [PSI] and [URE3] are reversibly curable. 2) [PSI] propagation requires SUP35 and [URE3] propagation requires URE2 with recessive chromosomal mutants having the same phenotypes as the presence of the respective dominant non-Mendelian element. 3) Overproduction of Sup35p and Ure2p increases the frequency of cells acquiring [PSI] or [URE3], respectively. PMID- 8720071 TI - Salivary mucins: protective functions in relation to their diversity. PMID- 8720072 TI - 1994, the year of sialyltransferases. PMID- 8720073 TI - Sialyl-Tn-KLH, glycoconjugate analysis and stability by high-pH anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD). AB - The quantitation of sialyl-Tn (STn) conjugated to keyhole limpet haemocyanin (KLH) can be determined by quantitating the amount of N-acetylneuraminic acid (NANA) released by acid or enzymatic digestion. An optimal 0.1 N H2SO4 acid hydrolysis at 80 degrees C results in quantitative release of NANA with minimal loss. A rapid isocratic method for the quantitation and separation of NANA is described using high-pH anion-exchange chromatography and pulsed amperometric detection (PAD). Multiple injection of NANA standard and/or samples containing protein led to a decrease in the PAD response which was corrected by addition of internal standard, alpha-2-keto-3-deoxyoctonate (KDO). The ratio of NANA/KDO peak area or peak height gives a linear response with increasing amount of NANA in the range 2.5-20 micro g/ml (r2 = 0.99). The limit of quantitation (LOQ) for NANA using this isocratic method is 1.9 micro g/ml (approximately 160 pmol/25 micro l injection). Based on the multiple determination the glycoconjugate, STn-KLH, showed a NANA content of 2.9% (w/w). Acid hydrolysis and the sialidase treatment of STn-KLH both yielded a similar NANA content. The carrier protein, KLH, showed the absence of NANA. The stability of glycoconjugate STn-KLH was monitored by a gradient method which separated possible degradation products STn-crotyl, NANA and GalNAc. Subjecting the glycoconjugate STn-KLH to various stress conditions of temperature, pH and oxidation does not result in any release of sialic acid, GalNac and STn-crotyl group. PMID- 8720074 TI - Strategy for the sequence analysis of heparin. AB - The versatile biological activities of proteoglycans are mainly mediated by their glycosaminoglycan (GAG) components. Unlike proteins and nucleic acids, no satisfactory method for sequencing GAGs has been developed. This paper describes a strategy to sequence the GAG chains of heparin. Heparin, prepared from animal tissue, and processed by proteinases and endoglucuronidases, is 90% GAG heparin and 10% peptidoglycan heparin (containing small remnants of core protein). Raw porcine mucosal heparin was labelled on the amino termini of these core protein remnants with a hydrophobic, fluorescent tag [N-4-(6-dimethylamino-2 benzofuranyl) phenyl (NDBP)-isothiocyanate]. Enrichment of the NDBP-heparin using phenyl-Sepharose chromatography, followed by treatment with a mixture of heparin lyase I and III, resulted in a single NDBP-linkage region tetrasaccharide, which was characterized as deltaUAp(1-->3)-beta-D-Galp(1-->3)-beta-D-Galp(1-->4)-beta Xylp -(1-->O-Ser-NDBP (deltaUAp is 4-deoxy-alpha-L-threo-hex-4-enopyranosyl uronic acid). Several NDBP-octasaccharides were isolated when NDBP-heparin was treated with only heparin lyase I. The structure of one of these NDBP octasaccharides, deltaUAp2S(1-->4)-alpha-D-GlcNpAc(1-->4)-alpha-L-IdoAp (1-->4) alpha-D-GlcNpAc6S(1-->4)-beta-D-GlcAp(1-->3)-beta-D- Galp(1-->3)-beta-D-Galp(1- >4)-beta-Xylp-(1-->O-Ser NDBP (S is sulphate, Ac is acetate), was determined by 1H-NMR and enzymatic methods. Enriched NDBP-heparin was treated with lithium hydroxide to release heparin, and the GAG chain was then labelled at xylose with 7-amino-1,3-naphthalene disulphonic acid (AGA). The resulting AGA-Xyl-heparin was sequenced on gradient PAGE using heparin lyase I and heparin lyase III. A predominant sequence in heparin at the protein core attachment site was deduced to be -D-GlcNp2S6S(or 6OH)(1-->4)-alpha-L-IdoAp2S-(1-->4)-alpha-D-GlcNp2S6S (or60H) (1-->4)-alpha-L-IdoAp2S(1-->4)-alpha-D-GlcNp2S6S( or 6OH)(1-->4)-alpha-L IdoAp2S(1-->4)-alpha-D-GlcNpAc (1- ->4)-alpha-L-IdoAp(1-->4)-alpha-D-GlcNpAc6S(1- >4)-beta-D-++ +GlcAp(1-->3)-beta-D-Galp(1-->3)-beta-D-Galp(1-->4)-beta-Xyl-AGA. PMID- 8720075 TI - alpha-galactosyl (Galalpha1-3Galbeta1-4GlcNAc-R) epitopes on human cells: synthesis of the epitope on human red cells by recombinant primate alpha1,3galactosyltransferase expressed in E.coli. AB - Developing methods for in vitro synthesis of the carbohydrate structure Galalpha1 3Galbeta1-4GlcNAc-R (termed the alpha-galactosyl epitope) on human tumour cells may be of potential clinical significance in cancer immunotherapy. Tumour vaccines with this epitope would be opsonized in vivo by the natural anti-Gal antibody, which is present in large amounts in humans, and which interacts specifically with alpha-galactosyl epitopes. Binding of anti-Gal to alpha galactosyl epitopes on tumour cell membranes is likely to increase uptake of the cell membranes by antigen-presenting cells, such as macrophages, via the adhesion of the Fc portion of anti-Gal to Fc receptors on these cells. This, in turn, may increase processing and presentation of tumour-associated antigens by antigen presenting cells, and induce an effective immune response against tumour cells with these antigens. The present study describes a method for the synthesis of alpha-galactosyl epitopes on human cells (red cells used as a model) by recombinant alpha1,3galactosyltransferase (rec. alpha1,3GT) expressed in bacteria. Escherichia coli was transformed with cDNA of the luminal portion of New World monkey rec. alpha1,3GT linked to six histidines (His)6 at the N terminus. The enzyme produced by the bacteria was isolated from bacterial lysates on a nickel-Sepharose column and eluted with imidazole. This recombinant enzyme displayed acceptor specificity similar to that of rec. alpha1,3GT produced in COS cells. Red cells were pre-treated with sialidase for exposure of N acetyllactosamine acceptors, then subjected to rec. alpha1,3GT activity. This enzyme synthesized at least 4 x 10(4) alpha-galactosyl epitopes/red cell. These epitopes were found to be accessible for binding of anti-Gal, as well as Bandeiraea simplicifolia IB4 lectin. It is argued that the method presented can be used for the synthesis of alpha-galactosyl epitopes on membranes of autologous tumour vaccines in humans. PMID- 8720076 TI - The backbone of the pectic polysaccharide rhamnogalacturonan I is cleaved by an endohydrolase and an endolyase. AB - Rhamnogalacturonan I (RG-I), a major pectic component of the primary walls of plant cells, is believed to play an important role in determining both the structure and functions of the walls. A more detailed structural description of RG-I is likely to lead to a greater understanding of the biological roles of this polysaccharide. Two enzymes secreted by Aspergillus aculeatus that have been cloned and expressed in a fungal system (Kofod et al., J. Biol. Chem., 269, 29182 29189, 1994) cleave the RG-I backbone in an endo fashion and should assist in the further structural characterization of this polysaccharide. We found that both of the available preparations of the cloned enzymes were contaminated with exoglycanases, reducing their utility in structurally characterizing RG-I. We purified the enzymes to apparent homogeneity by ion-exchange chromatography and then used the purified enzymes to generate backbone oligosaccharide fragments from partially debranched sycamore RG-I. The backbone oligosaccharides, which were separated from larger pieces of partially debranched RG-I by gel-permeation chromatography, have been structurally characterized by 1H-NMR spectroscopy, electrospray MS, GC-MS, high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD) and UV spectroscopy. The results of these analyses establish that rhamnogalacturonase A (RGase A) is an endohydrolase that cleaves the -4)-alpha-D-GalpA-(1-2)-alpha-L-Rhap glycosidic linkage. However, the purported rhamnogalacturonase B (RGase B) is, in fact, an endolyase that cleaves the -2)-alpha-L-Rhap-(1-4)-alpha-D-GalpA glycosidic linkage, thereby generating oligosaccharides terminating at the non-reducing end with a hex-4 enopyranosyluronic acid residue. PMID- 8720077 TI - Glycan structure of a heptose-containing S-layer glycoprotein of Bacillus thermoaerophilus. AB - The characterization of the S-layer glycoprotein of Bacillus thermoaerophilus revealed unexpected novelties. The isolation and purification procedure had to be changed due to complete solubility in aqueous buffers of the constituting S-layer protomers. Upon degradation of the S-layer glycoprotein by pronase and purification of the products by gel filtration, ion-exchange chromatography, chromatofocusing and HPLC, one representative glycopeptide fraction was selected for further characterization. From the combined evidence of composition analysis, chemical degradation, NMR spectroscopy experiments and comparison with synthesized model substance, we propose the following repeating unit structure of the glycan chain: -->4)-alpha-L-Rhap-(1-->3)-beta-D-glycero-D-manno-Hepp-(1--> This is the first description of heptose residues occurring as a constituent of S layer glycoproteins of gram-positive eubacteria. PMID- 8720078 TI - GlcNAc-transferase V and core 2 GlcNAc-transferase expression in the developing mouse embryo. AB - UDP-GlcNAc:Manalpha1-6Manbeta-R beta1-6-N-acetylglucosaminyltransferase V (GlcNAc TV) and UDP-GlcNAc:Galbeta1-3GalNAc-R beta1-6-N-acetylglucosaminyltransferase (core 2 GlcNAc-T) are Golgi enzymes that catalyse the biosynthesis of beta1 6GlcNAc-branched intermediates in the N- and O-linked biosynthesis pathways, respectively. The activities of these enzymes change during haematopoiesis, embryo-carcinoma cell differentiation and following malignant transformation, but little is known about their expression in normal adult tissues and during embryogenesis. We have examined the expression of GlcNAc-TV and core 2 GlcNAc-T in sections of post-implantation mouse embryos by in situ RNA hybridization. The two enzymes showed distinct temporal and spatial patterns of expression. Core 2 GlcNAc-T mRNA was widely expressed at embryonic day (E) 7, and became restricted to a subset of mucin- and cartilage-producing tissues at E11.5 through to E17.5. GlcNAc-TV transcripts were absent at E7, became expressed throughout E9.5 embryos, and then progressively restricted to regions of the developing central nervous system and to specialized epithelia of skin, intestine, kidney, endocrine tissues and respiratory tract. In the adult gonads, GlcNAc-TV transcripts were excluded from germ cells, but were detected in the follicular and testicular cells. Leukoagglutinin (L-PHA)-reactive oligosaccharides co-localized with GlcNAc TV transcripts in skin, kidney and intestine, but brain showed unexpectedly low overall staining punctuated by bright staining of the vascular endothelium. A common feature of cells in basal epithelia and in the cortical neural epithelium is the capacity to migrate, a cellular function which may require GlcNAc-TV dependent glycoconjugates. PMID- 8720079 TI - Identification of O-sulphate substituents on D-glucuronic acid units in heparin related glycosaminoglycans using novel synthetic disaccharide standards. AB - The two disaccharides, methyl 4-O-(2-O-sulpho-beta-D-glucopyranosyl-uronic acid) 2-deoxy-2-amino-alpha-D-glucopyranoside and methyl 4-O-(3-O-sulpho-beta-D glucopyranosyluronic acid)-2-deoxy-2-amino-alpha-D-glucopyranoside, were prepared by de novo synthesis, and converted to the corresponding 2,5-anhydro-D-[1 3H]mannitol derivatives by deamination with nitrous acid followed by reduction with NaB3H4. The resultant labelled products were used as standards in the identification, by anion-exchange high-performance liquid chromatography (HPLC), of disaccharides generated by HNO2/NaB3H4 treatment of heparan sulphate isolated from human brain. The two standards, containing 2-O- and 3-O-sulphated glucuronic acid, respectively, were clearly separated by the HPLC procedure. Comparison with the deamination products derived from heparan sulphate showed that the mono-O sulphated disaccharide species containing a sulphated glucuronic acid unit co eluted with the 2-O-sulphated standard. The corresponding component isolated from other heparan sulphate preparations, or from heparin, also eluted at the same position. No disaccharide derived from heparin or heparan sulphate appeared at the elution position of the 3-O-sulphated standard. It is concluded that D glucuronic acid units in heparin-related glycosaminoglycans may be sulphated at C2, whereas no evidence has been found for sulphation at C3. By contrast, analysis of mono-O-sulphated disaccharides derived from a chemically sulphated, bacterial capsular polysaccharide (generated by Escherichia coli K5) clearly demonstrated the occurrence of O-sulphate groups at C-3 of D-glucuronic acid units. PMID- 8720080 TI - Glycosylation and biological activity of CAMPATH-1H expressed in different cell lines and grown under different culture conditions. AB - CAMPATH-1H (where CAMPATH is a trade mark of Wellcome group companies), a humanized IgG antibody used in the therapy of lymphoma, leukaemia and rheumatoid arthritis, has been expressed in Chinese hamster ovary, Y0 myeloma and NS0 myeloma cell lines. These engineered cell lines were grown under different culture conditions, and the antibody isolated and purified. N-Linked oligosaccharides, on the CH2 heavy chain region of the antibody, were isolated and analysed by hydrazinolysis, high-performance anion-exchange chromatography with pulsed amperometric detection, laser-desorption mass spectrometry and sequential exoglycosidase treatment. Both the glycosylation pattern and the biological activity of CAMPATH-1H, as measured by antibody-dependent cell mediated cytotoxicity, were markedly affected by the cell line used to express the antibody. It is concluded that glycosylation of the antibody may be important in the clinical outcome of therapy. PMID- 8720081 TI - The natural history of renal artery stenosis: who should be evaluated for suspected ischemic nephropathy? AB - Ischemic renal disease is defined as a clinically significant reduction in glomerular filtration rate in patients with hemodynamically significant renal artery stenosis. The most common etiology for this is atherosclerotic renal artery disease. The three major clinical settings in which one must suspect ischemic renal disease include acute renal failure precipitated by the treatment of hypertension particularly with angiotensin-converting enzyme inhibitors; progressive azotemia in a patient with known renal vascular hypertension treated medically; and unexplained progressive azotemia in an elderly patient with refractory hypertension and other evidence of atherosclerotic disease. Prevalence of ischemic renal disease secondary to atherosclerosis can be estimated from the incidence of atherosclerotic renal artery lesions leading to renal vascular hypertension and the natural history of these lesions. Autopsy series, arteriography studies, and review of populations of patients in end-stage renal disease programs all suggest that ischemic renal disease has a high and increasing prevalence in our aging population. PMID- 8720082 TI - Pathogenesis of atherosclerosis. AB - Atherosclerosis, a common and complex disease, results from multiple interactions among injurious stimuli and the healing or reparative responses of the arterial wall. After endothelial injury, direct cell-cell interaction, and secretion of chemotactic and growth factors resulting from endothelial cell dysfunction, induce recruitment of monocytes to subintimal regions, smooth muscle cell proliferation, and increased synthesis of matrix proteins. The recruited monocytes become macrophages, accumulate lipid, and ultimately become foam cells. Together with accompanying T lymphocytes, these changes represent the fatty streak, an early histopathological change indicating atherosclerosis. Progression of this atherosclerotic lesion is marked by the accumulation of alternating layers of smooth muscle cells and lipid-laden macrophages. The advanced lesions of atherosclerosis compromise the lumen diameter and, thus, reduce the blood flow in arteries and ultimately participate in the mechanisms that lead to occlusion of the involved arteries. PMID- 8720083 TI - The pathology of chronic renal ischemia. AB - Chronic ischemia may cause end stage renal disease, especially in older patients with atherosclerotic renal artery stenosis. Examining the pathology of the ischemic kidney is a fundamental first step toward understanding the mechanisms of this injury. In experimental renal hypoperfusion, there is evidence of a mixture of adaptive responses, tubular and endothelial cell damage and repair events. These processes are reflected in a wide spectrum of morphological changes that include atrophy, focal necrosis, epithelial regeneration, apoptosis, inflammation, interstitial fibrosis, and thrombosis. The most severe damage is seen in the outer medulla, a region with marginal oxygenation even in normal circumstances. In the usual clinical case, the effects of aging, pre-existent hypertension, and the process of atherosclerosis further complicate the pathological picture. Lesions related to these factors include arteriosclerosis, athero-emboli, various types of glomerulosclerosis, and severe tubulointerstitial damage leading to "atubular glomeruli" and regional cortical scarring (nephrosclerosis). In this article, some mechanisms determining the varied and complex pathological findings that may be observed in individual cases are outlined. PMID- 8720084 TI - Cellular and metabolic consequences of chronic ischemia on kidney function. AB - Approximately 15% of end-stage renal disease is attributable to chronic ischemic nephropathy from renovascular disease, representing significant patient morbidity and sizable medical costs. Although the pathophysiology of both ischemic acute renal failure and renovascular hypertension are under intense study, there have been little data obtained on the pathophysiology of chronic ischemic injury to the kidney. Data from studies of renovascular hypertension demonstrate the primary dependence of the stenotic kidney on angiotensin II in maintenance of glomerular filtration rate, although other vascular regulators, such as endothelium-derived nitric oxide and endothelin, may also play a role. Clues to the pathophysiology of cellular injury in chronic ischemic nephropathy can be found in acute models of ischemic injury to the tubules, toxic models of chronic decreased blood flow such as cyclosporine, and from recent pathological studies showing immunologic alterations. Because there are very little data on the cellular mechanisms of chronic ischemic injury to the kidney, this is an important area for laboratory investigation, particularly because the techniques developed both in studies of acute renal ischemia and chronic renovascular hypertension are readily available. Further understanding of the cellular mechanisms of chronic renal ischemia may eventually lead to medical interventions for patients with ischemic nephropathy too ill to undergo major abdominal surgery. PMID- 8720085 TI - Ischemic nephropathy: noninvasive testing. AB - Ischemic nephropathy is defined as hypertension and a defect in renal function and/or loss of renal parenchyma caused by hemodynamic changes secondary to a renal artery stenosis. A work-up should be reserved for those patients considered candidates for intervention with percutaneous transluminal renal angioplasty (PTRA) or reconstructive surgery in whom there is hypertension with clinical features suggestive of ischemic nephropathy and/or a significant size disparity between the kidneys. Therefore, all patients suspected of ischemic nephropathy should undergo a renal ultrasound examination to quantitate renal size and a test of overall renal function (creatinine clearance or DTPA plasma disappearance). The diagnostic accuracy of functional tests for renovascular hypertension can be seriously impaired in the presence of renal insufficiency, and they have not been evaluated systematically in this clinical setting. Therefore, tests are reserved for those patients in whom the clinical clues of ischemic nephropathy are equivocal. Available evidence suggests that angiotensin-converting enzyme inhibitor (ACEI)-induced changes in the Hippuran or Mag3 renogram are predictive of reversible renovascular hypertension in the presence of renal insufficiency. Duplex ultrasound scanning of the renal arteries is an accurate method to identify renal artery stenosis in experienced hands. However, there is no quantitative information to judge the utility of noninvasive tests in predicting which patients with ischemic nephropathy will have an improvement in renal function after intervention. PMID- 8720086 TI - Options for therapy of ischemic nephropathy: role of angioplasty and surgery. AB - The development of chronic renal failure from progressive atherosclerotic renal arterial occlusive disease, termed "ischemic nephropathy," is an important clinical problem in older patients with generalized atherosclerosis obliterans. Studies on the natural history of atherosclerotic renal artery disease have made it possible to identify those patients in whom this disease poses a significant threat to overall renal function. Several factors must be considered in evaluating the renal functional benefit of intervention to relieve arterial obstruction. These include the severity and extent of renal artery obstruction, the level of renal function, renal histopathologic information, and the general medical condition of the patient. Intervention to restore normal renal arterial blood flow is indicated in selected patients to prevent deterioration of renal function that may culminate in the need for dialytic replacement therapy. Percutaneous transluminal angioplasty is the treatment of choice for patients with non-ostial atherosclerotic lesions. Most reports in the literature indicate that surgical revascularization provides more effective long-term therapy for patients with ostial atherosclerotic lesions. PMID- 8720087 TI - Estimates of mean daily intakes of persistent organochlorine pesticides from Spanish fatty foodstuffs. AB - In many countries pesticide residues in foods are monitored to ensure that public health is not endangered by residue daily intakes in excess of the recommended tolerance levels (van Dokkum and de Vos 1987). In Spain, there is only a total diet study carried out during 1971-72 by Carrasco et al. (1976). In that study, mean daily intakes of 11.5 mu g alpha-HCH, 13.8 mu g lindane and 78.4 mu g DDTs were calculated. Livestock meat and dairy products were the prime sources of human dietary exposure to organochlorines, since between 60-85% of the mean daily intakes arose from these particular food classes. These percentages are in accordance with the well documented fact that organochlorines predominantly accumulate in the lipid fractions of the human food chain, by which animal fatty foods have become a major route of exposure for humans (Kannan et al. 1992). Since the current daily intakes of organochlorines in Spain are not known, it was considered necessary to carry out a pesticide survey in several foods that compose an average Spanish diet. To accomplish that, we have determined residues of a list of priority organochlorine compounds in several fatty foodstuffs collected between 1987 to 1990, and prepared in the way in which they would normally be eaten. This study is merely an attempt to estimate the actual intakes, since only a selected number of food classes were investigated and no age-sex group, or seasonal differences were taken into account. In spite of these disadvantages, there are merits to such an approach. Approximate intake figures are available for comparison with toxicologically acceptable intakes and with retrospective studies in Spain and other countries around the world, and they serve to outline the temporal trends in organochlorine contamination that have occurred during the last decades. Also, it may contribute to diminish the consumer's concern about possible health risks involved in the consumption of food products and help to restore confidence in the quality of our food supply. PMID- 8720088 TI - Levels of cerium in the tissues of rats fed a magnesium-restricted and cerium adulterated diet. AB - Cerium, a lanthanide, is the most abundant rare earth element present in monazite and is biologically active. It has been postulated that cerium toxicity in conjunction with magnesium deficiency causes tropical endomyocardial fibrosis, a restrictive human cardiomyopathy (Valiathan et al. 1989; Valiathan and Kartha 1990). Generally, lanthanides are known to be poorly absorbed (Durbin et al. 1956; Evans 1990). The present study was carried out to ascertain whether magnesium deficiency promotes accumulation of cerium in various organs of rats. PMID- 8720089 TI - Metal binding and resultant loss of phototoxicity of alpha-terthienyl: metal detoxification versus alpha-terthienyl inactivation. AB - The members of the plant family asteraceae are noted for their cosmopolitan distribution and their versatility is attributed to be largely due to their morphological adaptations. Interestingly members of this family are endowed with rich levels of secondary plant metabolites, many of which are photochemically active (Bakker et al. 1979; Kagan et al. 1989). The secondary plant metabolite alpha-terthienyl derived from the plant family asteraceae is among the new class of light activated insecticide. The photobiocidal effects associated with alpha terthienyl in presence of sunlight and ultraviolet light (300-400 nm), has stimulated a great deal of interest in its toxic mechanism of action against a number of organisms including phytopathogenic fungi, nematodes and mosquito larvae. Trials under tropical conditions indicate a very high level of activity as a larvicide to mosquito. There is no cross resistance to this compound in malathion resistant mosquito larvae (Arnason et al. 1989). Even though many researchers feel that the phototoxicity of secondary plant metabolites has arisen independently many times in evolution as a defense mechanism, the physiological impact of such biologically active compounds in the plant producing them also should be addressed (Arnason et al. 1987). Moreover, the accumulation of secondary plant metabolites in the roots of many asteraceae members as is the case with alpha-terthienyl in marigold (Tagetes sp.) roots, hints towards their functional divergence. In this study, we demonstrate a potent heavy metal quenching activity of alpha-terthienyl. PMID- 8720090 TI - Effect of Ziram, Thiram, and Dithane M-45 on bone marrow cells of mice-assessed by micronucleus test. AB - Micronucleus test is an extensively used protocol to assess the mutagenicity of environmental chemicals. This was developed by Schmid and his co-workers (Matter and Schmid, 1971; Ledebur and Schmid, 1973). The micronucleus test is simple, quick and as sensitive as the chromosome aberration analysis. It is based on the principle that during anaphase, acentric chromatid and chromosome fragments lag behind, where as centric elements move towards the spindle pole. After telophase both the undamaged chromosomes and the centric fragments give rise to the daughter nuclei. The lagging elements are transferred into one or several secondary nuclei, which are as a rule much smaller than the main nucleus, and therefore called micronucleus (Schmid, 1973). The clastogenic effect of various chemicals is measured by micronucleus test. Erythrocytes are two types, the younger ones are polychromatic erythrocytes (PCE), which stain bluish and the older, the normo chromatic erythrocytes (NCE) which stain reddish. A few hours after the completion of last mitosis the erythroblasts expel their nucleus for unknown reasons and the micronucleus alone remains in the cytoplasm of the Polychromatic erythrocytes, and they are easily recognisable. Erythrocyte micronucleus represents the consequence of chromosomal aberrations induced during preceding mitotic division of erythrocytes (Matter and Grauwiler, 1974). PMID- 8720091 TI - Levels of PCDDs and PCDFs in human milk from populations in Madrid and Paris. PMID- 8720092 TI - Peridomestic deposition of ultra-low volume malathion applied as a mosquito adulticide. PMID- 8720093 TI - Seasonal atrazine contamination of drinking water in pig-breeding farm surroundings in agricultural and industrial areas of Croatia. AB - Atrazine (2-chloro-4-ethylamino-6-isopropylamino-s-triazine) a s-triazine herbicide, has been widely used in Croatian agriculture. Due to atrazine extensive use and its biodegradation in nature within at least one year (Klassen and Kodoum 1979), atrazine residues are found in ground, surface, drain and drinking water (Vidacek et al. 1994; Gojmerac et al. 1994). Groundwater downgradient from atrazine treated fields may show seasonal concentration peaks which could exceed the safe level (Wehtje et al. 1983). Therefore, the use of atrazine includes permanent control of its residues in water, particularly in relation to its use as a herbicidal chemical and groundwater contamination (Graham 1991). Furthermore, the presence of atrazine in the environment and its possible ingestion via the water, food and feed chain, may present a risk for the animal and human health. The analysis of atrazine residues in soil can be performed by either colorimetry or high performance liquid chromatography (HPLC) (Vickrey et al. 1980), and in water, soil and food by immunoassay in comparison with HPLC or gas chromatography/mass spectrometry (GS-MS) (Bushway et al. 1988; Bushway et al. 1989; Bushway et al. 1992; Thurman et al. 1990). We describe the use of enzyme-linked immunosorbent assay (ELISA) for one-year seasonal monitoring of atrazine residues in drinking water from two differently situated pig-breeding farms (agricultural and industrial areas) in Croatia. Results obtained by ELISA were compared to those produced by HPLC. PMID- 8720094 TI - Residues of dicloran in clingstone peaches after pre- and postharvest application. AB - Trials were carried out over a period of several years in order to test the efficacy of pre- and postharvest treatments against fungi causing postharvest rots of stored clingstone peaches destined to canning. Among the fungicides tested, dicloran (2, 6-dichloro-4-nitroaniline) was applied against Botrytis cinerea, Monilia cinerea and Rhizopus stolonifer. The purpose of this work was to determine the residues of this fungicide in fresh, washed, peeled and canned fruits. No relevant data are reported in the open literature to our knowledge. Some residue data, most of them produced by the manufacturer of the compound, were evaluated by the FAO/WHO Joint Meeting on Pesticide Residues (JMPR) and are summarized in the 1974 and 1977 monographs (FAO/WHO 1975 and 1978). PMID- 8720095 TI - Dissolved metal concentrations in surface waters from west-central Indiana contaminated with acidic mine drainage. PMID- 8720096 TI - Porphyrins as biomarkers of methylmercury and PCB exposure in experimental quail. AB - Chemicals such as heavy metals and polyhalogenated hydrocarbons have a high capacity to interfere with the enzymatic processes responsible for haem biosynthesis. These compounds can produce accumulation in tissues and organs and increased elimination of porphyrins in excreta (Andrew et al, 1990). The development of fast and easy analytical methods and the wide variety of biological media in which porphyrins can be detected have suggested their use as biomarkers of environmental pollution (Akins et al, 1993; De Matteis and Lim 1994). The analysis of porphyrins in the excreta is of special interest because it enables nondestructive monitoring of wild animals in the assessment of threatened or endangered species (Fossi et al, 1994). Methylmercury and PCBs are ubiquitous global pollutants and there is evidence they accumulate in terminal consumers, particularly those belonging to marine trophic chain (Renzoni et al, 1986; Yamashita et al, 1993). There have been some reports on methylmercury induced (e.g. Woods et al, 1991; Bowers et al, 1992; Miller and Woods 1993) and PCB-induced porphyria (e.g. Vos and Pennings, 1971; Miranda et al, 1987; Elliot et al, 1990; Miranda et al, 1992) but little data on their combined effect. In order to investigate the quality of porphyrins as biomarkers we performed an experiment in which Japanese quail were fed a diet containing methylmercury and polychlorobyphenyls (PCBs as Arochlor 1260) individually or combined in different ratios. The present study aims to provide preliminary data on liver and fecal levels of porphyrins in response to methylmercury and PCB administration, and on whether the indicator is sensitive to synergism or antagonism between the two compounds, administered simultaneously. PMID- 8720097 TI - Heavy metals in tissues of hares in Finland, 1980-82 and 1992-93. PMID- 8720098 TI - Volatilization of mercury by resting mercury-resistant bacterial cells. AB - The mercuric ion reduction system encoded by the Hg2+ inducible mer operon confers bacterial resistance to mercuric ion. The mer A gene product which is a FAD-containing enzyme catalyzes the reduction of Hg2+ to volatile elemental mercury with the help of intracellular thiols and NADPH as a cofactor (Schottel 1974; Summers and Silver 1978; Fox and Walsh 1982; Misra 1992). Our earlier studies have shown that growing cells of different mercury-resistant bacteria reduce Hg2+ compounds to Hg(O) (Ray et al. 1989; Pahan et al. 1990a; Gachhui et al. 1989). We have also shown the effect of thiol compounds and flavins on mercury-degrading enzyme activities in mercury-resistant bacteria (Pahan et al. 1990b). Here we report that resting cells of mercury-resistant bacteria survive in a buffer system for several hours, synthesize inducible mercury-degrading enzymes and volatilize mercury from a mercury-containing buffer system. We know of no information regarding studies of mercury-degrading enzymes in resting mercury-resistant bacterial cells. PMID- 8720099 TI - Chronic toxicity of phenanthrene to the marine polychaete worm, Nereis (Neanthes) arenaceodentata. AB - Polycyclic aromatic hydrocarbons (PAHs) are widely distributed in the environment. While environmental concentrations are generally below acutely lethal levels (Hyland and Schneider 1976), chronic, low level exposures may result in subtle sublethal effects. PAHs accumulate in bottom sediments and may represent a hazard to the benthos. Polychaetes are important members of this community (Officer and Lynch 1989). The objective of this study is to evaluate the chronic sublethal effects of one PAH, phenanthrene (PHN), on the polychaete worm, Nereis (Neanthes) arenaceodentata. PHN was selected because of its high toxicity to marine invertebrates relative to other PAHs (Neff 1979). PMID- 8720100 TI - Comparative sensitivity of three age classes of Artemia salina larvae to several phenolic compounds. AB - Phenolic compounds have been used because they are among the most toxic and ubiquitous environmental contaminants present in many industrial wastes. It is recognized that factors such as water hardness, pH, temperature, chemical formulation, species, age and/or stage of development of test organisms, to mention only a few, may critically affect the behavior of a chemical and thus effect the outcome of toxicity tests (Canton and Adema 1978; Berglind and Dave 1984; Persoone et al. 1989). Several studies dealing with the susceptibility of early life stages of invertebrates to pollutants have been reported in the literature (Middaugh and Dean 1977; Conklin and Rao 1978; Kaur and Dhawan 1993). An extensive literature review revealed information on the toxicity of different phenolic compounds to some species of fish and invertebrates. However, there is little information on acute toxicity of these compounds to different age classes of Artemia salina larvae. Artemia salina are of major importance in the aquaculture industry and they have been proposed as a uniform world-wide test system for toxicity of chemical substances (Vanhaeke et al. 1981) and for studies in developmental toxicology (Sleet and Brendel 1985). Previous investigations in this laboratory have shown that Artemia salina larvae exhibit increased sensitivity to certain chemicals in relation to aging. The present study was conducted to determine the acute toxicity of some phenolic compounds (pentachlorophenol (PCP), 2,6-dichloroindophenol (2,6-DCIP), 2,4-dinitrophenol (2,4-DNP), o-nitrophenol (o-NP), p-nitrophenol (p-NP), diamidophenol, diaminophenol and 2,6-dimenthylphenol (2,6-DMP) on Artemia salina 24-, 48- and 168- hr old to determine if changes in sensitivity occur during the first week after emergence. PMID- 8720101 TI - Assessment of the impact of naphthalene contamination on mangrove fauna using behavioral bioassays. AB - Pollution of the marine and estuarine environments by petroleum hydrocarbons is a world wide phenomenon (Connell and Miller 1980) and whilst large scale crude oil spills are the most obvious source of pollution, since the 1970's the impact of chronic, low level hydrocarbon input from sources such as oil refineries has been recognised as having long term ecological consequences, even when there may be no visible evidence of acute effects (Connell and Miller 1980). Mangroves are perhaps the dominant and most important intertidal habitat along subtropical and tropical coastlines and estuaries and as such are located in areas of high risk of acute or chronic petroleum hydrocarbon pollution. Further, once contamination occurs, high levels of hydrocarbons may be expected to remain in mangrove sediments as conditions are not favorable for hydrocarbon depletion by sediment transport or degradation by aerobic bacteria. Much research has focused on determining the acute toxicity of the water soluble fraction of crude or fuel oil to aquatic fauna but relatively little attention has been given to individual hydrocarbons. The medium to low boiling point aromatics such as naphthalene and its alkyl derivatives are the most toxic petroleum fraction to marine organisms (Anderson et al. 1974; Moore and Dwyer 1974) and they are known to provoke behavioral responses in marine animals at sublethal concentrations (Hargrave and Newcomb 1973; Linden 1977; Nagarajah et al. 1985). The goal of this investigation was to investigate the effects of a single aromatic petroleum hydrocarbon, naphthalene, in a subtropical mangrove environment, through the use of behavioral bioassays. The test organism chosen was the intertidal gastropod Ophicardelus quoyi, which is abundant in mangroves throughout eastern Australia. PMID- 8720102 TI - Estrogenic activity in rainbow trout determined with a new cDNA probe for vitellogenesis, pSG5Vg1.1. PMID- 8720103 TI - Biochemical and histopathological effects of glyphosate on carp, Cyprinus carpio L. AB - Glyphosate, also known by the trade names Roundup and Rodeo for agricultural use, is a broad-spectrum, translocated herbicide, used primarily in agricultural applications, and for vegetation control in non-crop areas. It is used as non selective herbicide and for aquatic weed control in fish-ponds, lakes, canals, slow running water, etc. (USDA 1984). Glyphosate is perhaps the most important herbicide ever developed. Literature of toxicological and ecotoxicological properties of glyphosate is extremely sparse, considering its importance as herbicide. Generally, glyphosate is slightly toxic to mammals and fish, but it may have an impact on the aquatic environment and also on the other aquatic organisms (USDA 1984). Due to this, its toxicity investigation is very important. The study of sublethal effects is of special importance for toxicological evaluation of compound. The objective of this study was to investigate acute and subacute toxic effects of sublethal glyphosate concentrations in water to carp (Cyprinus carpio L.), one of the commercially most important fish species populating freshwaters of Yugoslavia. PMID- 8720104 TI - Tributyltin oxide induced physiological and biochemical changes in a tropical estuarine clam. PMID- 8720105 TI - Electromyogram as a measure of heavy metal toxicity in fresh water and salt water mussels. AB - The response of bivalves to heavy metals and other toxins has usually been determined by observing valve position (e.g., Davenport and Manley 1958; Borcherding 1992). Since mussels close their valves to avoid noxious stimuli, experimental delivery of chemicals is uncertain. To obtain constant results, Preston (1994 and personal communication) employed plastic spacers to hold the valves apart. This obviates the observation of valve position as an index of response, and some other method is required. Electromyography of intact mussels is one such index, and is shown to be a simple, effective and quantitative measurement of activity. Experiments are reported on the effects of added mercury on salt water and fresh water species. Parts of this work have appeared in brief form (Kidder and McCoy 1995). PMID- 8720106 TI - Bioaccumulation and physiological changes in Hydrilla verticillata (l.f.) royle in response to mercury. PMID- 8720107 TI - Regulation of essential heavy metals (Cu, Cr, and Zn) by the freshwater prawn Macrobrachium malcolmsonii (Milne Edwards). AB - Despite the low concentrations of heavy metals in the surrounding medium, aquatic organisms take them up and accumulate them in their soft tissues to concentrations several fold higher than those of ambient levels (Bryan 1979; Rainbow et al. 1990). Knowledge of accumulation patterns of a particular trace metal is a prerequisite for understanding the significance of an observed metal concentration in a particular animal, especially from the aspect of biomonitoring. Many marine invertebrates accumulate heavy metals without any regulation and the accumulation necessarily being associated with mechanisms to store the metals in a detoxified form. Two detoxification mechanisms have been described, both of which may occur in one specimen. Heavy metals can either be bound up in insoluble metalliferous 'granules' (Mason and Nott 1981), or are bound to soluble metal-binding ligands, such as metallothioneins (Roesijadi 1992). Some marine decapod crustaceans have an innate ability to regulate the internal concentrations of essential but potentially toxic metals within a constant level, presumably to meet their metabolic demands (Rainbow 1985, 1992). However, at present, there is no such information relating to freshwater decapod crustaceans, especially shrimps which occupy a totally different environment. Macrobrachium malcolmsonii (Milne Edwards), a potential aquaculture species for freshwater is found in abundance in one of the major Indian rivers, the Cauvery. In the present study, an attempt was made to determine whether the freshwater prawn, M. malcolmsonii, is able to regulate the three essential elements, copper, chromium and zinc, over a wide range of dissolved concentrations. These three metals were chosen because the Cauvery River receives pollutants containing these metals (Vijayram et al. 1990). PMID- 8720108 TI - Sequence analysis of the conserved protamine gene cluster shows that it contains a fourth expressed gene. AB - Structural data are presented on the protamine gene cluster (PGC) of human, mouse, rat, and bull. By restriction mapping we demonstrate that the organization of the protamine cluster is conserved throughout all four species, i.e., the genes are situated in a head to tail arrangement in the order: protamine 1 protamine 2-transition protein 2. Further, we established the nucleotide sequence of the entire human PGC (25 kb in total) and the 3' portion of the rat protamine cluster (PRM2 and TNP2 genes and intergenic region). In addition, a 1 kb fragment of the bovine and murine protamine cluster, situated between PRM2 and TNP2, was sequenced. This fragment is conserved regarding sequence, position, and orientation in all species examined, and was classified as likely coding region by gene recognition program GRAIL. Using the rat fragment as a probe in RNA blots, we detected a testis-specific signal of about 0.5 kb. Finally, we demonstrate a high density of Alu elements, both full and fragmented copies, in the human PGC and discuss their localization with respect to evolutionary and functional aspects. PMID- 8720109 TI - Construction of a mouse blastocyst cDNA library by PCR amplification from total RNA. AB - Studies of the development and differentiation of early mammalian embryos have been severely limited by the paucity of material. Such studies have been largely restricted to the examination of abundant genes/proteins or to developmental expression studies of known genes for which DNA sequence data are available, allowing the use of reverse transcription and polymerase chain reaction amplification (RT-PCR). To eliminate the need for hundreds or thousands of oocytes or embryos in the construction of representative cDNA libraries, we describe a technique for generating and cloning cDNA using small caesium chloride gradient centrifugation to isolate total RNA from oocytes or embryos, followed by RT-PCR of mRNA from this total RNA. Total RNA was isolated from 70 mouse blastocysts. A portion of the cDNA generated (equivalent to seven blastocysts) was cloned, yielding a mouse blastocyst cDNA library of 1 million clones. We show that the library is representative in that it contains beta-actin, intracisternal A-type particles, tissue plasminogen activator, and B1 and B2 repetitive elements in frequencies comparable with published data from conventionally constructed libraries and estimates of mRNA abundance from expression studies. Furthermore, DNA sequencing of 22 clones chosen at random and compared with DNA sequence databases shows that approximately half are novel sequences. These data demonstrate that representative cDNA libraries can be constructed in situations where cell numbers are limiting and will facilitate the isolation of novel and interesting clones. PMID- 8720111 TI - Low levels of chimerism in rabbit fetuses produced from preimplantation embryos microinjected with fetal gonadal cells. AB - The potential pluripotency of rabbit fetal germ cells has been investigated by using them to make chimeric embryos. Gonial cells, isolated from enzyme-dispersed male and female transgenic fetal rabbit gonads of 18-22 days gestation, were microinjected in groups of about 10 into 640 nontransgenic rabbit embryos between the two-cell and expanded blastocyst stages. Injections were made with primary isolations of gonial cells, within 48 hr of their collection. The injected embryos were transferred, with or without non-injected control embryos, into 49 recipient rabbits. Tissues from 159 resulting fetuses, implantation sites, and a few liveborn young were examined by PCR analysis for the two transgenes used (alpha-1 antitrypsin or luciferase). The overall pregnancy rate (about 80%) was not affected by the stage of development of the embryo injected, nor by co transfer of control embryos. The survival rate of injected embryos (18% overall, 23.6% in pregnant recipients) was almost identical to that of 243 control embryos. Chimerism was detectable in tissues produced from 4 of 159 (2.5%) of the injected embryos, all four of which had been injected at the 8- to 16-cell stage. This low rate of success indicates that, although passage of rabbit gonial cells is not an absolute requirement for pluripotency, further investigation should pay particular attention to improving culture conditions with a view to deriving EG cell lines. PMID- 8720110 TI - Activation of the complete mouse metallothionein gene locus in the maternal deciduum. AB - The mouse metallothionein (MT) gene family consists of four known members (MT-I through IV) clustered on chromosome 8. Studies reported herein examine the expression and regulation of the MT-III and MT-IV genes in specific cell types in the maternal reproductive tract, developing embryo, and fetus known to express the MT-I and -II genes. MT-III and MT-IV mRNAs were absent from the visceral yolk sac, placenta, and fetal liver, tissues with high levels of MT-I and MT-II mRNAs. In contrast, MT-III and MT-IV mRNAs were both abundant in the maternal deciduum, and in experimentally induced deciduoma on 7 and 8 days postcoitum (1 dpc = vaginal plug), as are MT-I and -II mRNAs. The abundance of each of these MT mRNAs increased coordinately during development of the deciduum (6-8 dpc), and in situ hybridization localized MT-I, MT-III, and MT-IV mRNAs to the secondary decidual zone of the antimesometrial region on 8 dpc, where in some regions all of the cells were apparently positive. Thus, all of the known mouse MT genes are co expressed in at least some of the cells in the secondary decidual zone. Electrophoretic analysis of decidual MT suggested that the MT-I, -II, and -III isoforms are abundant proteins in the secondary deciduum. Bacterial endotoxin lipopolysaccharide (LPS) and Zn are powerful inducers of MT-I and MT-II gene expression in many adult organs, whereas these agents apparently have little effect on MT-III and MT-IV gene expression. Neither of these agents significantly effected levels of decidual MT-III or MT-IV mRNAs in vivo or in primary cultures of decidual cells in vitro, and only modest effects of Zn on MT-I mRNA levels were noted. During 2 days of in vitro culture, decidual cell MT-I and MT-III mRNA levels remained elevated while MT-IV mRNA levels decreased. Thus, expression of the mouse MT gene locus in the deciduum appears to be developmentally regulated, and in this tissue, the MT genes are refractory to induction by Zn or inflammation. PMID- 8720112 TI - Monoclonal antibodies against unfertilized zona-free mouse oocytes: characterization and effects on fertilization. AB - A panel of anti-oocyte antibodies was raised against unfertilized zona-free mouse oocytes by intrasplenic immunization and checked for their effects on in vitro fertilization. Four antibodies decreased the fertilization rate from about 90% in controls to 8% (B5-2 F7), 12% (A2-2 A7), 13% (4-G1), and 25% (A2-2 F2), when the sperm cell concentration was 1 x 10(5) to 1 x 10(6). Antigen localization: All the antibodies labelled components in the cell membrane of zona-free oocytes as demonstrated by indirect immunofluorescence and/or by complement-mediated oocyte lysis. In various patterns, the ooplasm and zona pellucida were also labelled with different intensities. Western blotting: A2-2 A7 and A2-2 F2 recognized a protein with a molecular weight of approximately 65 kDa, while antibody B5-2 F7 bound a 97 kDa protein. Complement activation and complement-mediated oocyte lysis: Systemically injected antibodies, C3 and C4 were detected on zona-free oocytes recovered from the mouse oviduct indicating the activation of C3 and C4 by antigen-antibody complexes. The recovered oocytes were not damaged, suggesting a presence of complement-regulating factors. In vitro, however, a large number of zona-free oocytes preincubated with antibodies were lysed or protruded ooplasma vesicles in complement-active serum. Stage, tissue, and species specificity: None of the antibodies, except A2-2 A7, showed a positive immunolabelling to the pronuclear stage. Antibodies 4-G1 and A2-2 F2 cross-reacted with the ovarian oocytes. No antibodies bound to any of the tissues tested, indicating that the corresponding antigen epitopes are not commonly expressed. A2-2 A7, A2-2 F2, and B5-2 F7 cross-reacted with hamster and human unfertilized oocytes, suggesting the presence of developmentally conserved molecules and the possibility to apply these antibodies in hamster and human in vitro fertilization. It is concluded that the approach used could be a useful strategy in searching for anti fertilization antibodies for human contraception. PMID- 8720113 TI - Cytofluorescent assay to quantify adhesion of equine spermatozoa to oviduct epithelial cells in vitro. AB - To facilitate the study of interactions between equine spermatozoa and homologous oviduct epithelial cells, we developed an assay to count labelled spermatozoa bound to oviduct epithelial cell (OEC) monolayers and used the assay to compare the binding ability of spermatozoa from different stallions. Washed spermatozoa from three stallions were incubated with the fluorochrome Hoechst 33342 (5 micrograms/ml) for 1 min. Spermatozoa were then layered over confluent monolayers of oviduct epithelial cells in 2 cm2 culture wells. Coculture treatments comprised five concentrations of spermatozoa (10(5), 5 x 10(5), 10(6), 2.5 x 10(6), and 5 x 10(6) per well). Cocultures were incubated for 30 min before unattached spermatozoa were aspirated in coculture supernatant. Fluorescent videoimages of attached spermatozoa were digitized, and attached spermatozoa were counted by image processing and analysis. Four wells (replicates) of each concentration were allocated within each ejaculate, and ejaculates were blocked by stallion for ANOVA. The total number of spermatozoa bound was not different between replicate wells (P > 0.1). Stallion, ejaculate, concentration, and all higher level interactions influenced total spermatozoa bound (P < 0.00001). Coefficients of variation between replicates were lowest for inseminate concentrations between 10(6) and 5 x 10(6) spermatozoa per well. Within the ejaculate, a log linear relationship exists between the number of bound spermatozoa and a spermatozoal concentration of the inseminate between 5 x 10(5) and 5 x 10(6) spermatozoa per well. This assay provides a reliable method of determining numbers of spermatozoa bound to somatic cells in vitro. Furthermore, differences exist in the ability of spermatozoa from different stallions to bind OEC monolayers. PMID- 8720114 TI - Detailed analysis of pronucleus development in bovine zygotes in vivo: ultrastructure and cell cycle chronology. AB - The ultrastructural development of pronuclei and cytoplasm was studied in bovine zygotes developed in the oviducts. The timing of the morphological events was related to sonographically detected ovulation and to the progress of the cell cycle determined by double labelling (3H and 14C-thymidine) of newly synthesized DNA combined with autoradiographic detection. The onset of the S-phase occurred at 11-12 hr after the estimated time of ovulation (EO), and this phase of the cell cycle lasted for 7-9 hr. During the G1-phase, the pronuclei contained spheres of compact, electron-dense fibrillar material classified as nucleolus precursor bodies. Early in the S-phase (13 hr after EO) spherical fibrillogranular bodies containing larger rounded electron-dense components were detected in the periphery of the pronuclei as well. At 15 hr, the latter bodies had become connected through electron-dense material with spherical multivacuolated fibrillar bodies of the same electron density as the nucleolus precursor bodies. At 17 hr, similar compact spherical bodies, now presenting a single large vacuole, were observed on some occasions, while in other zygotes the morphology remained unchanged throughout the rest of the S and G2-phases. PMID- 8720115 TI - Initial evaluation of fertilin as an immunocontraceptive antigen and molecular cloning of the cynomolgus monkey fertilin beta subunit. AB - Fertilin (PH-30) is a sperm surface protein that functions in sperm adhesion and fusion with the egg plasma membrane. Because of its essential function in fertilization, fertilin is a potential target for novel contraceptive approaches. In a pilot fertility trial, immunization of male guinea pigs with purified guinea pig fertilin resulted in complete infertility. The contraceptive effect was partial (two out of six animals were infertile) when female guinea pigs were immunized with the antigen. These results suggest that fertilin or domains of fertilin may be effective as immunocontraceptive antigens. As a step toward achieving this goal, we communicate the cDNA and deduced amino acid sequence of the monkey fertilin beta subunit. PMID- 8720116 TI - Maturity and fertility of rhesus monkey oocytes collected at different intervals after an ovulatory stimulus (human chorionic gonadotropin) in in vitro fertilization cycles. AB - In rhesus monkeys undergoing ovarian stimulation for in vitro fertilization (IVF), a midcycle injection of human chorionic gonadotropin (hCG) substitutes for the LH surge and induces preovulatory oocyte maturation. The time interval between injection and oocyte collection, ideally, allows for the completion of oocyte maturation without ovulation, which would reduce the number of oocytes available for harvest. To evaluate the influence of this time interval on oocyte parameters following hCG administration, we conducted a series of gonadotropin treatment protocols in 51 animals in which the interval from hCG administration to follicular aspiration was systematically varied from 27 to 36 hr. Follicle number and size, evaluated prior to hCG administration by sonography, did not vary significantly or consistently with preovulatory maturation time. Oocytes were harvested by laparotomy or laparoscopy, and scored for maturity before insemination. The percentage of mature, metaphase II (MII) oocytes at recovery increased significantly with increasing preovulatory time and was inversely proportional to that of metaphase I (MI) oocytes. However, oocyte yield tended toward a progressive decrease with increasing preovulatory maturation times from a high of 27 oocytes at 27 hr to a low of 17 oocytes/animal at the 36 hr time interval. Fertilization levels declined significantly from a high of 50% at 27 hr to a low of 30% at 36 hr. Thus, although higher percentages of mature oocytes were recovered at the longer time intervals, optimal oocytes/embryo harvests were realized after the shorter time intervals (27 and 32 hr) and are most compatible with the goal of achieving high yields of fertile oocytes and embryos following gonadotropin stimulation in rhesus monkeys. PMID- 8720117 TI - Properties and localization of a tyrosine phosphorylated form of hexokinase in mouse sperm. AB - Mouse sperm possess a phosphotyrosine-containing hexokinase type 1 (HK1) that is associated with the plasma membrane fraction of these cells (Kalab et al., 1994; J. Biol Chem 269:3810-3817). This apparent plasma membrane association appears unique, since somatic HK1 is normally cytoplasmic or bound to the outer mitochondrial membrane via contact sites with a voltage-dependent anion channel (porin) through a porin-binding domain. In male germ cells, three cDNA clones have been described that encode unique HK1 isoforms (HK1-sa, HK1-sb, HK1-sc) that do not contain porin binding domains (Mori et al., 1993: Biol Reprod 49:191-203). This suggests that these proteins might not be localized to the outer mitochondrial membrane and could have alternative functions in germ cells and/or sperm. We demonstrate in the mouse that male germ cells and sperm could potentially express four HK1 isoforms (HK1-sa, HK1-sb, HK1-sc, and the somatic HK1). At the protein level, at least one of the HK1 isoforms becomes phosphorylated on tyrosine residues during spermatogenesis. Treatment of sperm membrane fractions to dissociate the phosphotyrosine-containing HK1 (pY-mHK1) yields results demonstrating that pY-mHK1 has properties of an integral membrane protein. Indirect immunofluorescence using a monoclonal antibody to HK1 demonstrates specific staining both in the head and tail regions of sperm. Surface biotinylation of intact sperm followed by precipitation with either polyclonal HK1 antiserum or with avidin-Sepharose suggests that pY-mHK1 possesses an extracellular domain. These results suggest that mouse sperm contain at least one HK1 isoform that is present on the sperm head, has an extracellular domain, and behaves as an integral membrane protein. PMID- 8720118 TI - Sperm from mice carrying two t haplotypes do not possess a tyrosine phosphorylated form of hexokinase. AB - Mouse sperm contain a tyrosine phosphorylated form of hexokinase type 1 (HK1; Kalab et al., 1994: J Biol Chem 269:3810-3817) that has properties consistent with an integral plasma membrane protein. Furthermore, this tyrosine phosphorylated form of HK1 has an extracellular domain and HK1 is localized to both the head and flagellum of nonpermeabilized cells (Visconti et al., 1995c). We have characterized HK1 in mature sperm from sterile tw32/tw5 mice (mutant sperm) that have defects in motility and sperm-egg interaction (Johnson et al., 1995: Dev Biol 168:138-149). Immunoprecipitation of mouse sperm extracts with an antiserum made against purified rat brain HK1 demonstrates the presence of HK1 in mutant sperm. Various biochemical and immunofluorescence assays indicate that at least a portion of the HK1 present in these cells is an integral membrane protein with an extracellular domain located on the sperm head and flagellum. However, immunoblot analysis with anti-phoshotyrosine antibodies demonstrates that HK1 in mutant sperm is not tyrosine phosphorylated. Northern blot and RT-PCR analysis does not indicate any obvious abnormalities in the transcription of somatic or germ cell-specific HK1 isoforms in mutant testes, and RFLP analysis of recombinant mice indicates that no genes specifying HK1 isoforms are located on chromosome 17. We have mapped the locus responsible for the lack of tyrosine phosphorylation of HK1 mutant sperm to the most proximal (to the centromere) of the four inversions within the t haplotype. A male sterility factor is located in this same inversion (Lyon, 1986: Cell 44:357-363). Since the mutant sperm are unable to complete fertilization, there could be a relationship between sterility and the lack of tyrosine phosphorylation of HK1 in these mutant sperm. PMID- 8720119 TI - What is the relationship between the metabolism of preimplantation embryos and their developmental competence? PMID- 8720120 TI - Functional role of glycosphingolipids in cell recognition and signaling. AB - Glycosphingolipids (GSLs), cell type-specific markers which change dramatically during ontogenesis and oncogenesis, have been implicated as playing major roles in cellular interactions and control of cell proliferation in multicellular organisms. These functional roles have been partially clarified through two types of studies: (i) Studies of cell recognition mediated by (a) GSL-GSL interaction, (b) GSL-lectin interaction, and (c) GSL-dependent modulation of integrin receptor function. (ii) Studies on control of transmembrane signaling by GSLs and/or sphingosine (Sph) derivatives, with emphasis on effects of these compounds on: (a) signaling pathways initiated by tyrosine kinase-linked receptors; (b) signaling systems mediated by protein kinase C, MAP kinase, other kinases, or cytosolic Ca2+ concentration, leading to changes in cellular phenotypes such as motility, proliferation, differentiation, and apoptosis. PMID- 8720122 TI - Correlation among secondary structure, amyloid precursor protein accumulation, and neurotoxicity of amyloid beta(25-35) peptide as analyzed by single alanine substitution. AB - Structure-neurotoxicity relationships of amyloid beta (25-35) peptide were studied by replacing each amino acid with Ala. In contrast to the general tendency in hydrophobicity-toxicity relationships, replacement of Asn27 yielded a more hydrophobic but less toxic analog and that of Met35 gave a less hydrophobic but more toxic one. Sedimentation profiles and CD spectra indicated that peptide aggregation via intermolecular beta-sheet formation is essential for the neurotoxicity of amyloid beta (25-35) peptide. The correlation between neurotoxicity and amyloid precursor protein accumulation suggested that the latter is one of the pathways of the neuronal death caused by amyloid beta protein. PMID- 8720121 TI - A novel protein phosphatase-1 inhibitory protein potentiated by protein kinase C. Isolation from porcine aorta media and characterization. AB - A novel phosphorylation-dependent inhibitory protein (IP) of porcine aorta myosin light chain phosphatase (PA-MLCP) was purified to homogeneity from porcine aorta media. The molecular mass of IP was 20 kDa. IP phosphorylated by endogenous potentiating kinase (IP-K) inhibited not only PA-MLCP activity, but also that of the catalytic subunit of protein phosphatase-1. The amino acid sequence of a peptide derived from IP phosphorylated with IP-K, RHARVT*VK, shared one of the consensus sequences phosphorylatable by protein kinase C (PKC), where T* was phosphorylated. IP was phosphorylated by PKC and the phosphorylated product inhibited PA-MLCP as strongly as IP phosphorylated with IP-K. PMID- 8720123 TI - Variability in heat-induced fragmentation of a protein in the presence of dodecyl sulfate: the role of an intramolecular sulfhydryl/disulfide exchange. AB - alpha-Amylase from Aspergillus oryzae was investigated to better understand how disulfide-bonded proteins behave during denaturation with dodecyl sulfate. It was previously reported that the alpha-amylase, when denatured with dodecyl sulfate, forms two species, D1 and D2. In D1, the disulfide bonds remain intact, while in D2, there is a restricted single sulfhydryl/disulfide (SH/SS) exchange reaction. This phenomenon was re-examined as follows: electrophoretic analysis of fragments created with and without modification of a free SH group or disulfide-reduced SH groups; N-terminal sequence analysis of the fragments; reactivity of a free SH group with Ellman reagent. Data from the former two analyses showed that the variability in the electrophoretic patterns results from cleavage at Asp163 Cys164 or Asp282-Cys283, provided that these Cys residues are reduced. Different reactivities of a SH group in D1 and D2 and the appearance of a polypeptide with a nonnative SS pairing in D2 fragments confirmed that the different electrophoretic patterns result from an intramolecular SH/SS exchange. This rearrangement accounts for the variety of electrophoretic patterns observed with D2 amylase and is a result of the creation of a specific free Cys283. Specifically, the variety appears to be due to the breaking of either Cys150 Cys164 or Cys240-Cys283, which in turn leads to the labilization of Asp163-Cys164 or Asp282-Cys283, respectively. PMID- 8720124 TI - Stage-specific expression of the calmodulin gene in Plasmodium falciparum. AB - We studied the synthesis of calmodulin mRNA and of calmodulin during the asexual cell cycle of Plasmodium falciparum. The expression of the gene was low during the early stages of the erythrocytic asexual cycle. When the parasites were 28 h old the level of mRNA increased steadily to reach a peak at the age of 40 h. During the last 8 h of the cycle a constant decrease in the amount of calmodulin mRNA was observed. The synthesis of the protein followed the expression pattern of the mRNA, but without the final decrease. The average amounts of calmodulin measured were 2.5 +/- 0.9 ng/10(8) 14-h-old rings, 4.38 +/- 0.35 ng/10(8) 36-h old trophozoites, 11.8 +/- 1.45 ng/10(8) 44-h-old trophozoites, and 19.6 +/- 1.9 ng/10(8) 48-h-old schizonts. The in vivo stability of mRNA in different developmental stages was evaluated by pulse and chase experiments. The RNA of very mature trophozoites was quickly degraded, in contrast with the high stability shown by the RNA in other stages. The results here suggest that the calmodulin gene expression in Plasmodium does not follow a housekeeping pattern of expression such as in other eukaryotic cells, but that it is regulated at the level of transcription and RNA degradation. PMID- 8720125 TI - The role of SH and S-S groups in Bacillus cereus beta-amylase. AB - The properties of sulfhydryl (SH) and disulfide (S-S) groups in Bacillus cereus BQ10-S1 Spo III beta-amylase have been investigated to clarify their roles in the enzyme action. Two out of three cysteine residues in B. cereus beta-amylase were found to form an S-S bond, which was found to be located between Cys91 and Cys99 by the analysis of an S-S containing peptide. The replacement of the soybean beta amylase model around L3 loop 1 revealed that the S-S bond is located at the root of this flexible loop that moves between open and closed forms during catalysis. The analysis of fluorescence labeled peptides revealed that the remaining free SH group was Cys331. Modification of Cys331 with N-ethylmaleimide or p chloromercuribenzoic acid (PCMB) caused inactivation of the enzyme. The rate constants for the reactions were consistent with those of Cys343 in soybean enzyme. The binding affinity of the PCMB-modified enzyme to maltose was also decreased. These results indicate that the modification of Cys331, which exists as a free SH group in B. cereus beta-amylase caused inactivation by a similar mechanism to that in the case of Cys343 in soybean beta-amylase as assumed from the sequence homology. This cysteine residue has a common role in beta-amylases irrespective their origin. PMID- 8720126 TI - Expression of human metallothionein-2 in Escherichia coli: cadmium tolerance of transformed cells. AB - A genetic approach was undertaken to investigate the physiological roles of human metallothionein-2. A constructed expression plasmid, pEXPMTII, in which human metallothionein-IIA cDNA was inserted downstream of a tryptophan-lactose promoter, was used to transform Escherichia coli JM105 strain. Cadmium-binding metallothionein was successfully expressed in E. coli in the medium containing cadmium, while copper and zinc-metallothioneins were scarcely observed in copper- or zinc-containing medium. The amino acid composition and sequence of the biosynthesized cadmium-metallothionein were analyzed. The selectivity of metals bound to metallothionein and the stability of metal-binding forms of metallothionein in E. coli were discussed. In addition, cadmium, zinc, or copper resistance of the cells expressing metallothionein was examined. Cells transformed with the plasmid pEXPMTII and cultured in a medium containing cadmium exhibited tolerance only to cadmium. It was demonstrated that human metallothionein-2 functioned for cadmium detoxification in E. coli. PMID- 8720127 TI - Correlation between the differences in the free energy change and conformational energy in the folded state of hen lysozymes with Gly-Pro and Pro-Gly sequences introduced to the same site. AB - We suggested for the introduction of a prolyl residue into a protein that if the N-terminus residue is glycine, an unfavorable interaction in the folded state caused by the introduction of the prolyl residue can be substantially avoided by use of mutant lysozymes in which Gly-Pro and Pro-Gly sequences are introduced to positions 101-102 in the loop region of the lysozymes [Ueda, T., Tamura, T., Maeda, Y., Hashimoto, Y., Miki, T., Yamada, H., and Imoto, T. (1993) Protein Eng. 6, 183-187]. In order to determine whether or not the information obtained is applicable to other regions, we prepared mutant lysozymes with Gly-Pro and Pro Gly sequences at position 47, which is located in the beta-sheet, positions 70 71, which are located in the loop, positions 117-118, which are located in the beta-turn, and positions 121-122, which are located in the 3(10)-helix. The free energy changes of the native and mutant lysozymes for unfolding were determined at pH 5.5 and 35 degrees C. However, a mutant lysozyme with the Gly-Pro sequence was not always stabler than that with the Pro-Gly sequence at the same site. On the other hand, in order to determine whether or not strain caused by these sequences exists in the folded or unfolded state, the structures of these mutant lysozymes were determined by use of energy minimization. On comparison of the differences in the free energy change between the mutant lysozymes with Gly-Pro and Pro-Gly sequences at the same site with those in their total local conformational energies, it was found there is a good correlation between them. Therefore, it was suggested that the difference in total local conformational energy caused by the introduction of a Gly-Pro or Pro-Gly sequence could be estimated by use of the energy minimized structure. Moreover, the correlation indicated that the differences in the free energy change between Gly-Pro and Pro Gly lysozymes may be reflected by the differences in the total local conformational energies in their folded state. It was suggested that the energy levels in the unfolded states of mutant lysozymes with Gly-Pro and Pro-Gly sequences at the same site in a Gdn-HCl solution were almost identical. PMID- 8720128 TI - A 96-kDa glycyrrhizin-binding protein (gp96) from soybeans acts as a substrate for casein kinase II, and is highly related to lipoxygenase 3. AB - A 96-kDa glycyrrhizin (GL)-binding protein (gp96) was purified to apparent homogeneity from an aqueous extract of soybeans by means of successive DEAE cellulose column chromatography, gel filtration on Superdex 200pg, GL-affinity column chromatography, and ion-exchange chromatography on a Mono S column (HPLC). The protein was identified as a GL-binding protein since it specifically binds to [3H]GA. Moreover, it is a lipoxygenase (an enzyme that catalyzes the oxygenation of unsaturated fatty acids) since (i) it displays lipoxygenase (LOX) activity at pH 6.5; (ii) it is recognized on Western blot analysis by antibodies against LOX 1 and LOX-2; and (iii) the sequence of the N-terminal 21 amino acid residues (SNDVYLPRDEAFGHLKSSDFL) of a 42-kDa fragment (p42) proteolytically generated from gp96 is identical to a sequence of soybean LOX-3. In addition, GL, glycyrrhetinic acid (GA) and soyasaponin beta g slightly inhibited LOX activity of the purified gp96 fraction, whereas oGA (a GA derivative) greatly inhibited its activity. Furthermore, CK-II catalyzed phosphorylation of gp96 was stimulated significantly by GL at doses between 1 and 10 microM, but this phosphorylation was inhibited completely by 50 microM GL. All these results taken together suggest that (i) gp96 purified from soybeans as a GL-binding protein belongs to the LOX family; and (ii) triterpenoid saponins, including GL, are involved in the regulation of the activities of CK-II and LOXs in plants, such as soybeans and roots of liquorice, which contain large quantities of saponins. PMID- 8720129 TI - Some characteristics of the fluorescence lifetime of reduced pyridine nucleotides in isolated mitochondria, isolated hepatocytes, and perfused rat liver in situ. AB - By extensively examining the experimental conditions for time-resolved spectrophotometry of non-transparent light scattering systems, we demonstrated the feasibility of quantitative analysis of both the fluorescence lifetime and intensity of reduced pyridine nucleotides in living tissues, suspensions of isolated liver mitochondria, and hepatocytes, as well as hemoglobin-free perfused rat liver being used systematically for measurements. The fluorescence decay was analyzed by the maximum likelihood method with a 4-component decay model. The lifetime of NADH observed in mitochondria (mean: 2.8 +/- 0.2 ns) was much longer than that of the free form in an aqueous solution (mean: 0.43 +/- 0.01 ns), and it was characterized as a protein-bound form. The lifetime was not affected by either aerobic or anaerobic conditions nor by the energy state, though the intensity changed markedly. The decay curves of isolated hepatocytes under normal aerobic conditions were the same as those of isolated mitochondria, though cytosolic NADH and NADPH were superimposed. Under the conditions of "unphysiological" acidosis, the mean lifetime became about 1.5 times longer than that under normal conditions. With perfused liver, the relative contributions of cytosolic NADH and NADPH were determined by infusing lactate and tert butylhydroperoxide. Cytosolic NADH did not contribute to the overall fluorescence of pyridine nucleotides. In contrast, about 70% of the total fluorescence intensity was due to cytosolic NADPH, but its decay parameters were essentially the same as those of mitochondrial NADH. No free form of either NADH or NADPH was detected in the cytosolic and mitochondrial spaces. We concluded that the changes in fluorescence intensity observed under the various conditions can be simply explained by a change in the amount of reduced pyridine nucleotides in tissues, rather than by changes in the microscopic environment. The wide applicability of time-resolved fluorescence photometry to in vivo studies is well documented. PMID- 8720130 TI - DNA fragmentation induced in high-cell-density culture of primary rat hepatocytes is an active process dependent on energy availability, gene expression, and calmodulin. AB - We previously reported that internucleosomal DNA fragmentation, a biochemical feature of apoptosis, was induced spontaneously in high-cell-density culture of adult rat hepatocytes. To understand better the intracellular mechanism of the DNA fragmentation in this system, we have examined the effects of several inhibitors of specific intracellular functions on the DNA fragmentation. We found that the DNA fragmentation could be suppressed by treatment of the cells with inhibitors of mitochondrial respiration, KCN and CCCP, or a protein synthesis inhibitor, cycloheximide. We also demonstrated that calmodulin inhibitors, chlorpromazine and W-7, could suppress the DNA fragmentation. Together, these results lead us to conclude that the DNA fragmentation in hepatocytes cultured at high cell density is an active process dependent on energy availability, gene expression, and calmodulin, rather than a passive event resulting from necrosis. However, by analyzing the incidence of apoptotic morphology during culture, we found that only 2-3% of cells exhibited apoptotic morphology, while the incidence of DNA fragmentation was estimated to be much higher. Based on these results, we estimated that the DNA fragmentation may result from the incomplete progression of apoptosis or from the occurrence of another type of active cell death. PMID- 8720132 TI - Direct evidence of macrophage differentiation from bone marrow cells in the liver: a possible origin of Kupffer cells. AB - Controversy has surrounded origin and differentiation of tissue macrophages. We directly demonstrate the differentiation of bone marrow cells into macrophages in the liver in vivo using a cell-labeling fluorescence dye, PKH-26. Bone marrow cells labeled with PKH26 were intravenously injected into syngenic mice, and these cells were tracked by flow cytometric analysis. The majority of the labeled cells were detected only in the liver after 4 days. Interestingly, antigens specific for macrophage lineage cells (F4/80, Fc gamma RII, and CD14) were detected on the liver-accumulated cells only 4 h after the injection. The pattern of the antigen expression changed to that of Kupffer cells (F4/80+, Fc gamma RII+, Mac-1-) after 4 days and remained so thereafter. These labeled cells in the liver were esterase staining-positive and showed phagocytic activity at day 7. The number of labeled cells among the Kupffer cells in the liver increased with days after injection. This indicates that bone marrow cells accumulate in the liver and differentiate into liver macrophages on site. Roles of factors secreted from hepatocytes are also discussed. PMID- 8720131 TI - Phosphorylation of HS1, GAP-associated p190 and a novel GAP-associated p60 protein by cross-linking of Fc gamma RIIIA. AB - Human Fc gamma receptor IIIA (hFc gamma RIIIA) cDNA was introduced into mouse macrophage/monocyte cell line P388D1, and several stable cell clones expressing hFc gamma RIIIA were isolated. This facilitated the study of the biological function of Fc gamma RIIIA in monocytes/macrophages. The cloned cells showed the high phagocytic activity mediated by hFc gamma-RIIIA, while the original P388D1 cells did not. In order to examine the phosphorylation of proteins involved in hFc gamma RIIIA signal transduction, these receptors were stimulated by cross linking. The cross-linking of hFc gamma RIIIA induced a rapid increase in tyrosine phosphorylation of several proteins, including PLC-gamma 1, Syk, HS1, and p21rasGAP-associated p190 and p60 proteins. Immunoblotting with a polyclonal antibody specific for the GAP-associated p62 protein, which was originally found in fibroblasts and is homologous with an RNA-binding protein, revealed that the p60 phosphorylated after cross-linking of hFc gamma RIIIA seemed to represent a novel GAP-associated protein unrelated to the known GAP-associated p62 protein, which was also present in the P388D1 cells. PMID- 8720133 TI - Sequence-specific DNA recognition of the Escherichia coli Ada protein associated with the methylation-dependent functional switch for transcriptional regulation. AB - The Escherichia coli Ada protein, a suicidal DNA methyltransferase, is converted into a transcriptional regulator for methylation-resistance genes by the transfer of a methyl group from a DNA methylphosphotriester to its own Cys69 residue. Here, we report the DNA recognition mode and the functional switch mechanism of the N-terminal 16 kDa fragment of the Ada protein. NMR analysis has revealed that the segment from residues 102 to 123 forms a helix-turn-helix structure. A site directed mutagenesis study has shown that the second helix in the helix-turn helix structure plays a crucial role in specific recognition of DNA. These results imply that the sequence-specific interaction of the Ada protein with DNA occurs through the helix-turn-helix motif. NMR experiments on the methylated protein-DNA complex showed line broadening for the amide proton signals from the helix-turn-helix motif and for the protons in the vicinity of Cys69. In the case of the nonmethylated protein-DNA complex, signal broadening was observed only for protons from the helix-turn-helix. These findings suggest that the residues in the vicinity of Cys69 come into direct contact with the cognate DNA after methylation. We propose that the direct contact of this region is a major factor for the "switch" that converts the Ada protein from a nonspecific DNA binding form to a transcription factor. PMID- 8720134 TI - A structural study of the carboxyl terminal region of the human erythrocyte band 3 protein. AB - Two peptides derived from the carboxyl terminal region of the human erythrocyte band 3 protein were identified as fragments releasable from cell membranes on trypsin digestion. These peptides, Asn-880-Lys-892 and Ala-893-Val-911-COOH, however, were resistant to trypsin, unless the cell membranes had been treated with high concentrations of NaOH. This suggests that the carboxyl terminal region is located in situ within the native band 3 molecule. Unlike in the cases of other portions of the band 3 protein, such as Gly-647-Arg-656, Ser-731-Lys-743, and Tyr-818-Lys-826, the release of the carboxyl terminal region was not inhibited by pretreatment of erythrocytes with 4,4'-diisothiocyanostilbene-2,2' disulfonic acid (DIDS), indicating that there is no major structural difference in the carboxyl terminal portion between the outward and inward facing forms. The carboxyl terminal region of the band 3 protein has a negative charge cluster. In the middle of the negative charge cluster, consensus sequences, Val-Asp-X-X-X-Leu Asp-Ala-Asp-Asp and Thr-Phe-Asp-Glu (TFDE), were found in the carboxyl terminal regions of aquaporin CHIP and glucose transporter 1, respectively. The sequence, TFDE, exists in the highly amphipathic 11-residue sequence of glucose transporter 1, and this amphipathic sequence has been suggested to promote normal membrane insertion of polytopic membrane proteins such as glucose transporter 1 and serine chemoreceptor. The role of the carboxyl terminal region of the band 3 protein is discussed. PMID- 8720135 TI - Phosphorothioate antisense oligodeoxynucleotides capable of inhibiting hepatitis C virus gene expression: in vitro translation assay. AB - Phosphorothioate antisense oligodeoxynucleotides (S-ODNs) designed to hybridize to the 5' region of the hepatitis C virus (HCV) genome were evaluated as to their ability to inhibit HCV gene expression, using an in vitro translation system. Three effective regions were found to interfere with the translation of HCV RNAs. These regions were region A [nucleotides (nt) 124 to 153], region B (nt 100 to 123), and region C (nt 324 to 360). Further detailed evaluation of S-ODNs within each region allowed us to propose some HCV-specific antiviral agent candidates. Two of them, SMS16 (nt 328 to 347) and SMS17 (nt 326 to 345), caused over 90% inhibition of HCV gene expression when present in a less than fourfold molar excess; this effect was sequence-specific and dose-dependent. PMID- 8720136 TI - A 35 kDa mannose-binding lectin with hemagglutinating and mitogenic activities from "Kidachi Aloe" (Aloe arborescens Miller var. natalensis Berger). AB - A novel lectin was isolated from the leaf skin of "Kidachi Aloe" (Aloe arborescens Miller var. natalensis Berger) by sequential chromatographies on Sephadex G-25 gel filtration, DEAE ion exchange, and Superdex 75 gel filtration columns. The native lectin exhibited a molecular mass of about 35 kDa on both gel filtration on a Superdex 75 column and native-PAGE under nonreducing conditions. SDS-PAGE in the presence or absence of beta-mercaptoethanol revealed two distinct peptides with molecular masses of about 5.5 and 2.3 kDa, respectively, in addition to a major 9.2 kDa subunit, indicating the presence of a partially processed subunit. The N-terminal amino acid sequence of the intact subunit showed homology with that of snowdrop lectin. The native lectin showed hemagglutinating activity toward rabbit but not human and sheep erythrocytes, and specifically bound to mannose like snowdrop lectin did, indicating that the Aloe and snowdrop lectins are structurally and functionally similar proteins. In addition, the native lectin showed strong mitogenic activity toward mouse lymphocytes. PMID- 8720137 TI - Determination of urinary acetylpolyamines by a monoclonal antibody-based enzyme linked immunosorbent assay (ELISA). AB - A monoclonal antibody (mAb), ASPM-2, produced against N-(gamma maleimidobutyryloxy)-succinimide (GMBS)-conjugated polyamine spermine [Spm; Fujiwara et al. (1994) Histochemistry 102, 397-404] was used for the development of an enzyme-linked immunosorbent assay (ELISA) for acetylpolyamines (Ac-PAs) in human urine. The ELISA is based on the principle of competition between an analyte and Spm-glutaraldehyde-bovine serum albumin conjugate-coated polystyrene microtiter wells for the mAb, followed by immunoreaction with biotinylated anti mouse immunoglobulin and horseradish peroxidase-streptavidin. The ASPM-2 mAb showed strong immunoreaction with N1,N12-diacetylspermine (2Ac-Spm), N monoacetylspermine (Ac-Spm), and N1-acetylspermidine (N1-Ac-Spd), the EC50 values being 29, 50, and 51 microM, respectively, but no cross-reaction with other PA related compounds or amino acids. The method was used to measure urinary Ac-PA levels in healthy subjects and cancer patients, without pretreatment of the specimens, mean concentrations of 3.25 and 2.80 mumol per 24-h urine, respectively (as N1-Ac-Spd), being found. The ASPM-2 ELISA for N1-Ac-Spd, which is the PA most relevant to the analysis of human urine among the three Ac-PAs mentioned above, is specific and accurate, and can easily be used to analyze large numbers of specimens in parallel. It should thus have potential for studying the relationship between urinary N1-Ac-Spd levels and cancer. PMID- 8720138 TI - Subunit association of gamma-glutamyltranspeptidase of Escherichia coli K-12. AB - gamma-Glutamyltranspeptidase [EC 2.3.2.2] of Escherichia coli K-12 consists of one large subunit and one small subunit, which can be separated from each other by high-performance liquid chromatography. Using ion spray mass spectrometry, the masses of the large and the small subunit were determined to be 39,207 and 20,015, respectively. The large subunit exhibited no gamma-glutamyltranspeptidase activity and the small subunit had little enzymatic activity, but a mixture of the two subunits showed partial recovery of the enzymatic activity. The results of native-polyacrylamide gel electrophoresis suggested that they could partially recombine, and that the recombined dimer exhibited enzymatic activity. The gene of gamma-glutamyltranspeptidase encoded a signal peptide, and the large and small subunits in a single open reading frame in that order. Two kinds of plasmid were constructed encoding the signal peptide and either the large or the small subunit. A gamma-glutamyltranspeptidase-less mutant of E. coli K-12 was transformed with each plasmid or with both of them. The strain harboring the plasmid encoding each subunit produced a small amount of the corresponding subunit protein in the periplasmic space but exhibited no enzymatic activity. The strain transformed with both plasmids together exhibited the enzymatic activity, but its specific activity was approximately 3% of that of a strain harboring a plasmid encoding the intact structural gene. These results indicate that a portion of the separated large and small subunits can be reconstituted in vitro and exhibit the enzymatic activity, and that the expressed large and small subunits independently are able to associate in vivo and be folded into an active structure, though the specific activity of the associated subunits was much lower than that of native enzyme. This suggests that the synthesis of gamma glutamyltranspeptidase in a single precursor polypeptide and subsequent processing are more effective to construct the intact structure of gamma glutamyltranspeptidase than the association of the separated large and small subunits. PMID- 8720139 TI - Dephosphorylation of fetal-tau and paired helical filaments-tau by protein phosphatases 1 and 2A and calcineurin. AB - We have reported that many sites of tau in fetal brain (fetal-tau) as well as in paired helical filaments (PHF-tau) are phosphorylated. In the present study, we used site-specific antibodies and peptide mapping to examine protein phosphatases involved in dephosphorylation of fetal-tau and PHF-tau. Immunoblot analysis and electrophoretic mobility showed that protein phosphatases 1 and 2A and calcineurin could dephosphorylate fetal-tau and PHF-tau. Phosphoserines 199, 202, 396, and 413 and phosphothreonine 231, numbered according to the longest human tau isoform, were dephosphorylated, as shown by the immunoblot analysis. Phosphoserine 422 was dephosphorylated by protein phosphatase 2A and calcineurin, but not by protein phosphatase 1. Peptide mapping with Achromobacter lyticus protease 1 showed that phosphoserines 199, 202, 235, and 396 and phosphothreonine 231 were dephosphorylated by protein phosphatases. Fetal-tau was more rapidly dephosphorylated by protein phosphatase 2A and calcineurin than PHF-tau. Interestingly, PHF-tau which had not been solubilized with guanidine HCl was little dephosphorylated by protein phosphatases. Thus, PHF-tau in neurofibrillary tangles of Alzheimer's disease brain is likely to be resistant to dephosphorylation by protein phosphatases. PMID- 8720140 TI - Actin-actin contact: chemical cross-linking between actin and the 2.6-kDa peptide from subdomain 4 of actin. AB - Previously, we demonstrated that the 2.6-kDa peptide extending from Arg177 to Tyr198 in subdomain 4 of rabbit skeletal actin bound to actin itself, inhibited the elongation of actin filament, and severed F-actin. The corresponding segment in actin, therefore, is thought to contain the most critical actin-actin contact [Hori, K. and Morita, F. (1992) J. Biochem. 112, 401-408; Hori, K., Itoh, T., Takahashi, K., and Morita, F. (1994) Biochim. Biophys. Acta 1186, 35-42]. In this paper, we report on the binding site in actin for the 2.6-kDa peptide studied by using a zero-length cross-linker, 1-ethyl-3(3-dimethylaminopropyl)carbodiimide (EDC). We conducted limited digestion of actin cross-linked with the 125I-labeled 2.6-kDa peptide with various proteases, and developed peptide maps. The cross linked region of the 2.6-kDa peptide was found to be within the region of Ala114 to Glu167 in actin by identifying the radioactive peptide fragments. The region was further restricted by isolation of radioactive peptide from alpha chymotryptic digest of the cross-linked actin. The binding site of the 2.6-kDa peptide was finally assigned to be within the 24 amino acid segment from Ala144 to Glu167, which lies in subdomain 3 of actin. Using computer graphics, actin actin contact provided by the two segments was suggested to be along the left handed genetic helix of actin filament. PMID- 8720141 TI - Crosslinking of a 28-residue N-terminal peptide of actin to myosin subfragment 1. AB - The N-terminal 28-residue peptide of actin whose Cys10 was labeled with 5 iodoacetamido-fluorescein (F3K peptide) was isolated from the fluorescently labeled thrombin digest of actin. The effect of myosin subfragment 1 (S1) on the fluorescence of F3K peptide was examined in the absence of ATP. With increasing concentration of S1 added, the fluorescence intensity of F3K peptide increased by maximally 7.3% with an apparent dissociation constant of 5.7 microM, suggesting a role of this peptide region of actin in acto-S1 binding in rigor. F3K peptide was crosslinked with S1 at 10 mM NaCl using a zero-length crosslinker by the method of Grabarek and Gergely [Anal. Biochem. 185, 131-135 (1990)]. The crosslinking was greatly inhibited by the presence of either 0.2 M NaCl or 5 mM MgATP. The analyses of amino acid compositions and sequences of the fluorescent peptides isolated from a lysylendopeptidase digest of the crosslinked S1 indicated that F3K peptide was mainly crosslinked to residues 637-642 of the S1 heavy chain. The crosslinked S1 was isolated by selectively pelleting the uncrosslinked S1 with F actin. ATPase activity of the isolated crosslinked S1 alone was twice as high as that of control S1. The actin-activated ATPase activity of the crosslinked S1 was much lower than that of uncrosslinked S1. The estimated Vm and Km values were 1.72 s-1 and 125 microM, respectively. The Vm decreased to less than 1/8, while Km increased only twofold. The results suggest that the N-terminal 28-residue segment of actin may be implicated in the rigor binding of actomyosin and in the actin-activation of myosin ATPase, but may not be the main determinant of actomyosin binding in the presence of ATP. PMID- 8720142 TI - Conformational aberrance of the sendai virus F0 protein in thapsigargin-treated cells allowing exit from the endoplasmic reticulum but causing arrest at the Golgi complex. AB - Thapsigargin and ionomycin inhibited the intracellular transport of the Sendai virus F0 protein. Depletion of Ca2+ from intracellular Ca2+ store(s) is critical for the inhibition, since ionomycin was more effective in the absence of extracellular Ca2+ than in its presence. Transport of F0 was arrested between the trans-Golgi complex and the plasma membrane [Ono, A. and Kawakita, M. (1994) J. Biochem. 116, 649-656], but only when thapsigargin was added before the synthesis of F0. This implies that F0 was committed to later arrest in the endoplasmic reticulum. Non-reducing SDS-polyacrylamide gel electrophoresis revealed a conformational abnormality of F0 immediately after pulse-labeling in thapsigargin treated cells. The abnormality did not affect the exit of F0 from the endoplasmic reticulum, but paralleled its later arrest at the trans-Golgi stage. Pulse labeled and 1-h-chased F0 was endoglycosidase H-resistant even in thapsigargin treated cells, but was not recognized by mAb f-49, a monoclonal antibody that recognizes the corresponding F0 intermediate in uninhibited cells. The misfolded F0 may escape from recognition by means of the quality control system of the endoplasmic reticulum, but another system in the Golgi complex may complement the former. The arrested F0 was rapidly degraded. PMID- 8720143 TI - Importance of 6-O-sulfate groups of glucosamine residues in heparin for activation of FGF-1 and FGF-2. AB - Treatment of the pyridinium salts of heparin with N-methyltrimethylsilyl trifluoroacetamide (MTSTFA) in pyridine for 2 h at various temperatures caused specific 6-O-desulfations from trisulfated disaccharide units to various degrees without detectable depolymerization or other chemical changes. In order to assess the importance of 6-O-sulfate groups in N-sulfated glucosamine (GlcNS) residues to promote FGF-1 and FGF-2 activities, various 6-O-desulfated (6-O-DS-) heparins were quantitatively examined for activity as enhancers or inhibitors of specific FGF-1- and FGF-2-induced proliferation of BALB/c3T3 clone A31 (A31) cells and the chlorate-treated cells. The present results suggested that a high content of 6-O sulfate groups in GlcNS residues was required for activation of FGF-1, but not FGF-2. However, complete 6-O-desulfation of trisulfated disaccharide units in heparin resulted in loss of the ability to activate FGF-2, although the desulfated product bound strongly to FGF-2. PMID- 8720144 TI - Orotate phosphoribosyltransferase from Thermus thermophilus: overexpression in Escherichia coli, purification and characterization. AB - Orotate phosphoribosyltransferase (OPRTase, EC2.4.2.10) plays a role in de novo synthesis of pyrimidine nucleotide and transfers orotate to 5-phosphoribosyl-1 pyrophosphate (PRPP) to form orotidine-5'-monophosphate (OMP). To obtain heat stable OPRTase and to elucidate the mechanism of heat stability, this enzyme from Thermus thermophilus was expressed in Escherichia coli and purified. The pyrE gene of T. thermophilus which encodes OPRTase, contains an open reading frame of 549 base pairs with 69% G+C content. Since this gene expressed itself inefficiently in E. coli, the 5' and 3' ends of the coding regions were replaced with synonymous codons which contain more A+T and corresponds to major codons for E. coli. Introduction of the modified gene fragments into a plasmid having a tac promoter resulted in production of a polypeptide of molecular weight (M(r)) 20,000 in the presence of isopropyl-beta-D-thiogalactopyranoside (IPTG) in E. coli. This protein represented as much as 16% of the bacterial total protein and showed the OPRTase activity. Three purification steps, consisting of heat treatment at 65 degrees C, 40% ammonium sulfate fractionation, and KCl gradient elution from DEAE-Sephadex A-50, resulted in highly purified single polypeptide. The optimum activity of the purified OPRTase was observed at 150 mM KCl, pH 9.0, 75-80 degrees C, and in the presence of 100 microM PRPP. The activation energy of this enzyme reaction was 20.3 kJ/mol. The Km of this enzyme for orotate as a substrate was 75 microM and the maximum specific activity was 300 units/mg protein under the optimum conditions. The purified OPRTase was stable for 20 min at 85 degrees C. PMID- 8720145 TI - Tissue-type transglutaminase is not a tumor-related marker. AB - The level of transglutaminase (TGase) expressed in various tumor cell lines was investigated. We found that each cell line could be categorized into three distinct groups, that is, (i) cell lines with low or negligible TGase activity, (ii) cell lines with significant or high TGase activity and immunoreactive to a monoclonal antibody against tissue type TGase, and (iii) cell lines with significant or high TGase activity but not immunoreactive to the antibody. Results reported here argue against the former proposal that tissue-type TGase is expressed at a lower level in malignant cells compared to the level in normal cells. PMID- 8720146 TI - Purification and characterization of rat brain transglutaminase. AB - Transglutaminase (TGase) catalyzes an acyl-transfer reaction between peptidyl glutamine residues and primary amines including the epsilon-amino group of lysine residues in protein. TGase in the neuronal system has been suggested to be involved in neurotransmitter release, long-term potentiation, and so forth. In order to study the mammalian brain TGase at the molecular level, TGase was purified to apparent homogeneity from Sprague-Dawley rat brain, using DEAE ion exchange, and heparin and alpha-casein affinity column chromatographies. The brain TGase was concentrated 11,400-fold and had a specific activity of 11,000 nmol/h/mg protein. The purified protein migrated on SDS-PAGE to a position corresponding to a molecular size of approximately 75 kDa. The brain TGase activity was Ca(2+)-dependent (EC50 approximately 0.28 mM), and its Km values for putrescine and N,N-dimethylcasein were 0.26 and 0.065 mM, respectively. GTP inhibited the brain enzyme activity 100-fold more potently than ATP did, and the enzyme was photolabeled with [alpha-32P]8-azido-GTP, suggesting that the brain TGase is a member of the GTP-binding protein family. Monoiodo-acetate and cystamine potently inhibited the enzyme activity, suggesting that cysteine residue(s) are essential for brain TGase. Zn2+ inhibited the enzyme, while Mg2+ was not inhibitory. The rat brain TGase reacted only weakly with monoclonal and polyclonal antibodies against tissue-type TGase such as guinea pig liver TGase and human red blood cell TGase, while it did not react with antibodies against non-tissue types of TGase, namely human epidermal TGase and human coagulation factor XIIIa. The results suggest that rat brain TGase shares major characteristics with tissue-type TGase, but is nevertheless distinct from tissue type TGase. PMID- 8720147 TI - Significance of the C-terminal domain of Erwinia uredovora ice nucleation-active protein (Ina U). AB - Ice nucleation-active (Ina) proteins of bacterial origin comprise three distinct domains, i.e., N-terminal (N-), central repeat (R-), and C-terminal (C-) domains, among which the R-domain is essential, and its length may be correlated with the ice nucleation activity. In addition, the short C-terminal domain of about 50 amino acid residues is indispensable for the activity. Using the Ina U protein of Erwinia uredovora, we carried out precise mutational analyses of its C-terminus. The ice nucleation activity (T50) assay showed that the C-terminal 12 amino acids were not necessary, and a deletion mutant (delta C29) with a new C-terminal, Met29 (numbered from the first amino acid residue of the C-domain and corresponding to Met1022), exhibited almost the same activity as the wild-type Ina U protein did. However, deletion of the C-terminal 13 residues including Met29 resulted in almost complete loss of the activity. In the deletion mutant (delta C29), amino acid replacement of the C-terminus, Met29, showed that the activity was retained when Met29 was replaced with a neutral, aromatic, or basic amino acid (Gly, Phe, or Lys), but was lost on the replacement with an acidic amino acid (Asp or Glu). In addition, two other residues in the C-terminal region commonly present in all Ina proteins were examined as to their importance, and it was shown that one of these residues, Tyr27, is important for the activity, although it is not exclusively required; the activity was lost to a great extent when this residue was replaced with Gly or Ala, but to a lesser extent when it was replaced with Leu. These results suggest that significance of the secondary and/or tertiary structure of the C-terminal region of the Ina U protein for the ice nucleation activity. PMID- 8720148 TI - Activation of smooth muscle alpha-actin promoter in ras-transformed cells by treatments with antimitotic agents: correlation with stimulation of SRF:SRE mediated gene transcription. AB - Smooth muscle alpha-actin promoter is repressed in ras-transformed fibroblast cells and derepressed in revertant cells. We have recently shown that serum response factor (SRF) which can bind to the serum response elements (SREs) present in the alpha-actin promoter, can activate alpha-actin promoter activity in ras-transformed cells and suppress transformation by ras. Agents that stimulate SRF expression and alpha-actin promoter activity in ras-transformed cells are expected to be potential candidates as antitumor agents. In this study, we show that treatment of ras-transformed cells with antitumor agents such as taxol, vincristine, vinblastine, colchicine, and nocodazole leads to 5- to 7-fold activation of alpha-actin promoter driven CAT activity, whereas there was very little effect on thymidine kinase promoter driven CAT activity. This activation occurred at subcytotoxic concentrations of these agents and correlated with inhibition of cell cycle progression. Furthermore, these agents stimulated SRF expression in ras-transformed cells, as measured by its SRE binding activity. The increase in alpha-actin expression is accompanied by the restoration of actin filaments into organized bundles. These results suggest a novel mechanism by which antimitotic agents suppress the ras-transformed phenotype. PMID- 8720149 TI - Methanol traps the troponin-tropomyosin-actin complex in an "off-state". AB - We have investigated the effect of methanol at concentrations below 15% (v/v) on acto-heavy meromyosin (HMM)- and tropomyosin (Tm)-troponin (Tn)-acto-heavy meromyosin-ATPase activities. Methanol slightly enhanced ATPase activity of acto HMM alone. It had no apparent effect on normalized Tm-Tn-acto-HMM-ATPase activity in the absence of Ca2+. In the presence of Ca2+, however, methanol markedly inhibited normalized Tm-Tn-acto-HMM ATPase activity. These results show that methanol affects the troponin-tropomyosin regulation of acto-HMM ATPase: methanol suppresses the Ca(2+)-sensitivity of the regulatory system and traps it in an "off-state." PMID- 8720150 TI - Peripherally biased distribution of antigen proteins on the recombinant yeast derived human hepatitis B virus surface antigen vaccine particle: structural characteristics revealed by small-angle neutron scattering using the contrast variation method. AB - The internal structure of the recombinant yeast-derived human hepatitis B virus surface antigen vaccine particle was investigated by small-angle neutron scattering using the contrast variation method. Data were collected in aqueous buffer solutions containing 0, 40, 60, and 100% D2O in the q range of 0.005 to 0.2 A-1 at 5 degrees C. The radius of gyration at infinite contrast and the maximum dimension of the particle were estimated to be 107 and 290 A, respectively. The contrast matching point of the particle was determined to correspond to about 30% D2O, indicating that a considerable portion of the vaccine particle is made up of lipids and carbohydrates from yeast. The distance distribution function of the particle at 40% D2O, at which the protein components are matched out to show only the distribution of the remaining lipids and carbohydrates, differed markedly from the functions at other D2Os, and showed the nearly symmetrical profile characteristic of a spherical particle with a diameter of 240 A. The Stuhrmann plot and profile of the distance distribution function at 40% D2O showed that (i) the vaccine is a spherical particle with a diameter of 290 A, in which two different regions in terms of scattering density are distributed radially, (ii) the lipids and carbohydrates form a spherical cluster with a diameter of 240 A in the core region of the particle, and (iii) the surface antigen protein is present in the peripheral region. This architecture of the vaccine particle is favorable for the induction of anti-virus antibodies. PMID- 8720151 TI - Localization and in vitro mutagenesis of the active site in the Saccharomyces cerevisiae mRNA capping enzyme. AB - The yeast mRNA capping enzyme is composed of 52 (alpha) and 80 kDa (beta) polypeptides, which are responsible for its mRNA guanylyltransferase and RNA 5' triphosphatase activities, respectively. We isolated the gene encoding the alpha subunit (CEG1) and showed that CEG1 is essential for yeast cell growth [Shibagaki et al., (1992) J. Biol. Chem. 267, 9521-9528]. In this study, CEG1 was expressed in Escherichia coli and the alpha subunit protein was purified to near homogeneity. A [32P]GMP-bound tryptic peptide derived from the recombinant enzyme [32P]GMP covalent reaction intermediate was converted to a [32P]phosphoryl peptide through periodate oxidation followed by beta-elimination. Hydrolysis of the [32P]phosphoryl-peptide with alkali resulted in [32P]N epsilon-phospholysine as the only phosphoamino acid, indicating that GMP in the enzyme-GMP complex is bound to a lysine residue via a phosphoamide linkage. Microsequencing of the [32P]GMP-peptide showed that the GMP binding site was located in the region between amino acids 60 and 75, which contained an internal trypsin-resistant lysine at position 70. CEG1 was subjected to site-directed mutagenesis and the mutant proteins were expressed in E. coli. Substitution of His or Ile for Lys70 entirely abolished the enzyme-GMP formation activity, and this mutation was lethal to yeast in vivo, supporting the notion that the active site in the alpha subunit is located at Lys70. Replacement of Lys70 with Arg reduced the ability to form the enzyme-GMP complex; however, yeast cells bearing this allele were not viable. A series of mutations, including 8 amino acid replacements and 3 insertions, near the active site (Lys70-Thr-Asp-Gly motif) were also introduced and the mutant polypeptides were examined for catalytic activity in vitro as well as yeast cell viability in vivo. There was a good correlation between the in vitro and in vivo functions of the mutant proteins, except when Asp72 was replaced with Glu, which allowed formation of the enzyme-GMP complex but failed to support cell growth. The results with Lys70 to Arg and Asp72 to Glu substitutions indicated that guanylyltransfer to RNA and/or additional roles besides cap formation per se are impaired in these mutant proteins. PMID- 8720152 TI - Strategic partnership: the successful contract laboratory-sponsor relationship. AB - Contract laboratory. These two words often trigger a number of concerns for a sponsor, concerns that range from timing and cost considerations to issues of technical and regulatory capability. From a contract laboratory perspective, the best way to address these concerns is for the sponsor to consider the contract laboratory as a strategic partner, a provider of needed services. In many ways, the lab should function as an extension of the sponsor's own technology group. This partnership approach should foster effective communication so that the contract laboratory understands both the technical and regulatory objectives of the sponsor as well as any unique factors that could impact the study. A team approach will ensure an understanding of the role of each participant, approaches to be used in problem solving, and details of the sponsor/laboratory interaction, such as protocol input/development, technical updates/reviews, quality assurance audits, and final report format and approval process. This presentation will share perspectives from one contract laboratory on strategic partnerships for good science, with compliance, remembering that for the sponsor, the successful study is done on time, on budget, and is accepted by the agency. PMID- 8720153 TI - Basic training in good laboratory practices for researchers. PMID- 8720154 TI - A quality assurance manager's perspective on in-life inspections. PMID- 8720155 TI - International agency inspections in Japan. PMID- 8720156 TI - The French GLP and GCP inspections and agencies plus an overview of GLP and GCP inspections and agencies in Eastern and Central Europe. PMID- 8720157 TI - Preparing for and handling agency inspections for the United Kingdom Department of Health. AB - This paper indicates some known differences in approach adopted by the United Kingdom Department of Health (DoH) Good Laboratory Practice (GLP) Monitoring Authority from that of other agencies and offers guidance to quality assurance (QA) professionals in dealing effectively with a DoH inspection. The UK GLP Programme contains, at present, just over 100 member laboratories, ranging from one-man pathology outfits to major companies with perhaps over 1000 staff working in compliance. Laboratories in the GLP program are subject to routine inspection every 2 years; without a satisfactory inspection and continued membership in the GLP program, UK laboratories cannot make a formal claim of compliance. Routine inspections occur at 2-year intervals, with a formal period of notice, allowing some preparation to occur. Routine inspections comprise the selection of a small number of studies for audit and inspection of all the facilities. Inspectors look critically at QA and management systems and routinely interview staff to get a feel for the skills, knowledge, competence, and attitude of those who work on nonclinical studies. Study directors are constant targets for the inspectors' inquisition. Throughout the inspection process, inspectors are accompanied by senior QA staff, whose notes are used to ensure that nothing in the final meeting is reported incorrectly or out of context. The inspectors leave after verbal discussion with management of any significant findings. A full written report is typically provided within 3 weeks, to which a written response from management is required within 4 weeks. There is no Freedom of Information program in the UK. PMID- 8720158 TI - Japanese authority GLP inspections for pharmaceuticals and agrochemicals. Preparing for and handling agency inspections. AB - This presentation covers the essentials of recent Good Laboratory Practice (GLP) inspections conducted in Japan, after applications have been made for new ethical drugs to be authorized by the Ministry of Health and Welfare (MHW) or for agricultural chemicals by the Ministry of Agriculture, Forestry, and Fishery. Recently, the functions of MHW/GLP inspections were, for the most part, transferred from MHW to "The Organization for Drug ADR Relief, R&D Promotion and Product Review" ("Drug Organization," in short). This organization is now responsible for the audit and facility inspections for GLP compliance, both for new drug applications and for animal safety tests conducted at contract laboratories. The methods of inspection are expected to be the same as those used by MHW. A recent inspection audited by the Ministry of Agriculture, Forestry, and Fisheries is also reviewed here, for comparison. PMID- 8720159 TI - Preparing for and handling the results of regulatory inspections of biomedical facilities. AB - Although there have been numerous presentations on how to conduct and survive a government inspection, it appears that these sessions do not always adequately include suggestions from those who have been the subjects of regulatory inspections. This paper will include comments from the experience of one of the regulated. Remarks apply to experience with U.S. regulatory authorities, principally the U.S. Food and Drug Administration (FDA) and the U.S. Environmental Protection Agency (EPA). This paper attempts to give guidance on how to prepare for an inspection and what to do at the debriefing and after the inspector(s) leave your facility. PMID- 8720160 TI - The role of QC/QA in "paper" NDA submissions. PMID- 8720161 TI - Beyond computer validation - a new role for quality assurance in systems development. AB - The Quality Assurance Unit (QAU) at Eli Lilly and Company is involved in the development of a new clinical pathology computer system in conjunction with the information systems area and the user area. This is the first time that the QAU had the opportunity to build our requirements into a new computer system. This is a new role for QA and includes both auditing and validation personnel who offer QA insights into issues. Benefits of QA involvement in the developmental process include saving time by building quality checks into the system, thus eliminating the need for manual review later in the process. Early resolution of issues leads to significant financial savings. Other benefits include improved communication among the three functional areas and a better understanding of the requirements and work processes. The team approach to the development of a new computer system results in a high-quality final product which meets all user and regulatory requirements, while also providing additional benefits to our organization. PMID- 8720162 TI - Enhancement of paraquat toxicity by glutathione depletion in mice in vivo and in vitro. AB - Effect of glutathione (GSH) depletion on paraquat (PQ) toxicity in the liver and kidneys of mice was examined. Glutamic-pyruvate transaminase (GPT) and blood urea nitrogen (BUN) levels in plasma of mice were hardly changed by treatment with 150 micro mol/kg of PQ. However, significant increases in the plasma GPT and BUN levels after PQ injection were observed in mice which were pretreated with L buthionine-SR-sulfoximine (BSO), an inhibitor of GSH synthesis, at 4 hr prior to PQ administration. This result supports the previous observation that hepatotoxicity of PQ was enhanced in diethyl maleate-pretreated mice (Cagen and Gibson, 1977). In the present study, lipid peroxidation evaluated by thiobarbituric acid-reactive substances (TBA-RS) level in the liver of mice given PQ was elevated by pretreatment with BSO. Moreover, enhancement of PQ cytotoxicity by BSO pretreatment was also observed in cultured mouse hepatoma cell line (NCTC clone 1469). Vitamin E, an antioxidant, and Desferal, an iron chelator, significantly prevented mice from the BSO-enhanced hepato- and nephrotoxicity of PQ. These findings suggest that the tissues or cells of low GSH concentration are highly vulnerable to PQ toxicity and GSH may play a major role in diminishing the toxic action of PQ exerted through oxidative stress. PMID- 8720163 TI - Functional human hepatocytes: isolation from small liver biopsy samples and primary cultivation with liver-specific functions. AB - Morphologically and functionally intact human hepatocytes were isolated from small liver biopsy samples weighing about 1-2 g by initial digestion with collagenase followed by repeated digestions with trypsin. The usual yield of hepatocytes was greater than 1 x 10(7) cells per g of liver sample and cell viability, as judged by dye exclusion test, was routinely over 90%. The isolated human hepatocytes showed intact morphology under scanning electron microscope. Formation of membrane protrusions upon phalloidin addition demonstrated that the actin in isolated hepatocytes was maintained with its structural integrity. The cultured human hepatocytes retained a variety of liver-specific functions which were similarly exhibited by rat hepatocytes isolated using the same procedure. The cultured human hepatocytes exhibited a specific cytochrome P-450 related enzyme activity, and active amino acid uptake that increased upon addition of hormones like glucagon and dexamethasone. Additionally, the cultured human hepatocytes synthesized DNA actively and, human serum albumin, and was found to be responsive to modulation by growth modulating hormones, cytokines and hepatotoxic agents. Based on the profile of activity described above, the presently established conditions for isolation and culturing of human hepatocytes demonstrate that functional liver cells can be obtained from small biopsied liver samples. PMID- 8720164 TI - Experimental methods for immunization and challenge in antigenicity studies in guinea pigs. AB - Optimal experimental methods for antigenicity studies in guinea pigs were investigated on: (1) the effects of different immunizing methods using complete or incomplete Freund's adjuvants (CFA or IFA), and various injection sites, the number of immunizations, the immunizing doses, and the immunizing periods, (2) the relationship between the severity of active systemic anaphylaxis (ASA) reactions and passive cutaneous anaphylaxis (PCA) titers, (3) positive control for oral administration, and (4) the effects of incubation mixture of drug and serum protein as the challenge for the ASA assay. The following results provided useful information for designing more appropriate methods for antigenicity studies: (1) The optimal immunization method for benzylpenicillin (PcG), cephaloridine, 2,4,6-trinitrobenzene sulfonic acid and adriamycin, which were selected as positive controls for low molecular medicines in this experiment, involved subcutaneous administration of 1 ml of a test substance in CFA (1st immunization) or IFA (2nd and 3rd immunizations) at two doses, 1 and 10 mg/animal, 3 times at 2-week intervals on the back of a guinea pig. Blood collection for PCA assay was needed 2 weeks after the last immunization, and ASA assay, 1 or 2 days after the blood collection. (2) The insensitivity of ASA reactions in bovine serum albumin-immunized animals with very high PCA titers was overcome by increasing the challenge antigen dose from 1 to 10 mg/animal. (3) Most animals administered lysozyme at 0.1, 1 or 10 mg/animal by gavage for 2 weeks or more showed ASA and PCA reactions. (4) Incubation of a mixture of 20 mg/ml of PcG and 2 mg/ml of guinea pig serum albumin for 4 hr was the most effective as challenge for the induction of ASA reaction in PcG-immunized guinea pigs. PMID- 8720165 TI - Delayed effects of ethanol, caffeine and nicotine assessed by wheel-running and drinking in mice. AB - Effects of ethanol, caffeine and nicotine, pleasurable substances, on wheel running and drinking in mice that were housed under a 12 : 12-hr light-dark schedule (lighting period ; 6 : 00-18 : 00) were investigated. All drug administrations were carried out at 11: 00, a mid-light period. Although ethanol (0.8-2.4 g/kg p.o.) scarcely changed both the wheel-running and drinking during the light period, it was followed by a strong suppression of both behaviors during the coming dark period (18 : 00-6 : 00). The same treatment with caffeine (1-10 mg/kg s.c.) produced significant increase in the drinking during the light period, but suppression of the wheel-running during the dark period. Nicotine (0.1-1 mg/kg s.c.) significantly suppressed the wheel-running, but not drinking, during the dark period. The coadministration of nicotine (0.1-1 mg/kg) with ethanol (2.4 g/kg) reduced the behavioral suppression during the dark period. Whereas nicotine (0.1-1 mg/kg) reduced the increased drinking during light period by caffeine (10 mg/kg), but enhanced the caffeine-induced behavioral suppression during the dark period. These results indicate that the administration of pleasurable substances in the mid-light period results in a delayed effect which is characterized by a suppression of either and/or both wheel-running and drinking during the coming dark period starting 7 hr after the administration, and that nicotine acts to antagonize the effect of ethanol, but contrally to enhance the effect of caffeine. PMID- 8720166 TI - Influence on HMG-COA reductase inhibitors on plaque-forming cell (PFC) response in mice. AB - Effects of pravastatin and simvastatin on the humoral immune response were studied using Cunningham's method of the IgM plaque-forming cell (PFC) assay system in mice. The vehicle (0.5% CMC), pravastatin (1 mg/body : about 50 mg/kg) or simvastatin (0.5 mg/body : about 25 mg/kg) were given orally for 14 days in experiment Exp-I and Exp-II. The vehicle, pravastatin (0.5 mg/body : about 25 mg/kg) or simvastatin (0.25 mg/body : about 12.5 mg/kg) were given orally 29 times for 32 days in Exp-III. There were no significant differences in the mean numbers of PFCs per 10(6) splenocytes between the pravastatin groups and the vehicle groups. In the simvastatin groups, however, there were significant reductions of the numbers of PFCs in Exp-I, Exp-II and Exp-III, as compared with the vehicle group. Based on these results, it was considered that suppressive effect on IgM antibody production was induced by simvastatin, whereas such a suppression was not observed with pravastatin. PMID- 8720167 TI - Effects of various post-treatment by phenylmethylsulfonyl fluoride on delayed neurotoxicity induced by leptophos. AB - Delayed neurotoxicity induced by leptophos, an organophosphorus insecticide, was intensified in hens when phenylmethylsulfonyl fluoride (PMSF) at dose of 30, 60, and 120 mg/kg body weight was administered at different time intervals (24 hr, 3 days, and 5 days) for each dose of PMSF after the hens were exposed to 30 mg/kg (i.v.) of leptophos. The scores for organophosphorus-induced delayed neuropathy (OPIDN) in all groups treated with 120 mg/kg PMSF were significantly higher than those in the group treated with leptophos only (P<0.05 or P<0.01) and the initial signs of OPIDN appeared 2 or 3 days earlier in the former groups than in the latter group. Further, the greater the PMSF post-treatment dose, the more severe were the signs of OPIDN. These findings indicate that post-treatment with PMSF promotes leptophos-induced OPIDN and reduces the period to OPIDN onset. We also examined the effects of various time intervals between PMSF administration and exposure to leptophos on the development of OPIDN. The OPIDN scores in the two groups of hen treated with PMSF on days 3 and 5 after leptophos exposure were high, especially the score of the 5 days treated group became significantly higher on the 18th and 19th day after leptophos administration than even that of the 24 hr treated group with PMSF (P<0.05). These findings suggest that variations in both the dose of PMSF and the time intervals of PMSF post-treatment may affect the delayed neurotoxicity induced by leptophos. Moreover, these results also indicate that PMSF should not be used for either the treatment or the prevention of OPIDN. PMID- 8720168 TI - Mutagenicity studies with palm fruit carotene. AB - The mutagenicity of palm fruit carotene was examined using the reverse mutation test with bacteria, the chromosomal aberration test with mammalian cells and the micronucleus test in mice. The carotene induced neither reverse mutation in Salmonella typhimurium TA98, TA1537, TA100, TA1535 and in Escherichia coli WP2uvrA, nor structural and numerical (polyploidy) chromosomal aberrations in the Chinese hamster fibroblast cell line (CHL). In addition, no increase in micronucleated polychromatic erythrocytes was elicited in the micronucleus test in CD-1(ICR) male mice. It is concluded that palm fruit carotene had no mutagenic activity in these in vitro and in vivo tests. PMID- 8720169 TI - [XXXVII annual meeting of Japan Society of Smooth Muscle Research (I). Abstracts]. PMID- 8720170 TI - Role of 3'-untranslated regions in the regulation of hexose transporter mRNAs in Trypanosoma brucei. AB - Trypanosoma brucei is a unicellular parasite that is transmitted from one mammalian host to the next by tsetse flies. The expression of many trypanosome genes is regulated during the life cycle but there is no evidence for developmental control of transcription by RNA polymerase II. T. brucei expresses at least two hexose transporter mRNAs that are developmentally regulated; we show here that specific portions of the 3'-untranslated regions are responsible for the differential expression. Different trypanosome 3'-untranslated regions, from surface protein, phosphoglycerate kinase and aldolase genes as well as the hexose transporter genes, conferred a spectrum of levels of reporter gene expression, and these activities differed between bloodstream forms and the procyclic forms that replicate in the tsetse vector. Experiments with permanently transformed cell lines showed that regulation occurs at the mRNA level. The results suggest that post-transcriptional control of mRNAs in trypanosomatids operates at several levels, and that it will not always be possible to attribute all the regulation to short RNA motifs. PMID- 8720171 TI - Amino acid uptake and intracellular accumulation in Leishmania major promastigotes are largely determined by an H(+)-pump generated membrane potential. AB - Leishmania major promastigotes maintain a relatively high pool of free amino acids (> 100 mM) under in vitro growth conditions. They also maintain a hyperpolarized plasma membrane which is primarily set by a dicyclohexylcarbodiimide (DCCD)-sensitive electrogenic H(+)-pump. We studied here the possible contribution of the membrane potential (Vm) and the transmembrane proton gradient (delta pH) to the mediated uptake of amino acids and their intracellular accumulation. Proline transport and accumulation were assessed by analysis of time-dependent changes in the internal pools of free amino acids and by uptake of radiolabelled proline. Proline uptake was markedly affected by changes in the Vm and considerably less by changes in delta pH. The most pronounced effects were obtained by treatment with either the H(+)-uncoupler carbonylcyanide chlorophenylhydrazone (CCCP), the cation ionophore gramicidin or by omitting Cl- from the medium (by exchange with gluconate or mannitol). Relatively smaller effects were obtained in the presence of the H(+)-ATPase inhibitor DCCD or with the anion transport blocker 4,4' diisothiocyanodihydrostilbene-2,2'-disulfonic acid (H2DIDS). No significant effects were found with cells exposed to K+ in the presence of nigericin, to Na+ in the presence of monensin or to other cations substituting for Na+. These results suggest that neither extracellular Na+ or K+, per se, nor even intracellular pH, play a major role in proline uptake and accumulation. A significant stimulation in proline uptake induced by HCO3- could be associated with membrane hyperpolarization or intracellular alkalinization. The present observations indicate that uphill nutrient uptake by Leishmania promastigotes is largely determined by Vm. The relatively high intracellular pools of amino acids might be of physiological relevance to osmoregulation by parasites. PMID- 8720172 TI - Expression of mammalian cytokines by Trypanosoma cruzi indicates unique signal sequence requirements and processing. AB - A vector based upon the calmodulin-ubiquitin 2.65 locus of Trypanosoma cruzi has enabled the expression and secretion of the murine cytokines interleukin-2 (IL-2) and gamma-interferon (gamma-IFN) by transfected T. cruzi. The T. cruzi-derived cytokines were bioactive and produced by both epimastigotes and mammalian forms. The native coding sequence of IL-2 was sufficient to cause secretion of the protein, but the gamma-IFN signal sequence had to be replaced by the IL-2 signal sequence (IL-2/gamma-IFN) to allow efficient secretion of gamma-IFN. The amino acid sequences at the N-termini of the secreted T. cruzi-derived cytokines were different from the expected murine secreted protein. The secreted IL-2 was cleaved six amino acids downstream from the murine signal sequence cleavage site, and the hybrid IL-2/gamma-IFN molecule was cleaved three amino acids downstream from the predicted signal cleavage site in the IL-2/gamma-IFN molecule. These apparent differences in signal peptide sequence requirements and cleavage sites most likely indicate that the signal sequence processing in trypanosomes is distinct from that of higher eukaryotes. PMID- 8720173 TI - Recombinant Pfs230, a Plasmodium falciparum gametocyte protein, induces antisera that reduce the infectivity of Plasmodium falciparum to mosquitoes. AB - Six regions of malaria transmission-blocking target antigen, Pfs230, encoding 80% of the 363-kDa protein, were expressed as recombinant proteins in E. coli as fusions with maltose-binding protein (MBP). Antisera generated against amylose purified recombinant Pfs230/MBP fusion proteins (r230/MBP.A-r230/MBP.F) all recognized the 360-kDa form of parasite-produced Pfs230 by immunoblot. However, only antisera against the four carboxy regions (C-F) of Pfs230 and not the two amino regions (A and B) recognized the 310-kDa form of Pfs230, the form expressed on the surface of gametes. The data suggest that the 310-kDa form of Pfs230 arises from the cleavage of 50 kDa from the amino terminus of the 360-kDa form. Furthermore, antisera against r230/MBP.C bound to the surface of intact gametes and significantly reduced (by 71.2-89.8% (rank sum analysis, P < 0.01)) the infectivity of P. falciparum parasites to mosquitoes. This is the first report of a recombinant form of a P. falciparum gametocyte protein capable of inducing antisera that reduce malaria parasite infectivity to mosquitoes. PMID- 8720174 TI - Characterization of the respiratory chain of Leishmania donovani promastigotes. AB - Inhibition analysis of respiration of Leishmania donovani promastigotes in resting, starved and permeabilized cells in the presence of classical electron transfer complex inhibitors such as rotenone, thenoyltrifluoroacetone and antimycin demonstrated the absence of complex I component of the respiratory chain in this organism. Cyanide failed to completely block the oxygen uptake (residual 25-30%) even at high concentrations. The alternative oxidase inhibitor for Trypanosoma brucei, salicylhydroxamic acid (SHAM) had no effect on respiration while the cytochrome o inhibitor orthohydroxydiphenyl (OHD) could block cyanide-insensitive respiration at low concentrations. Succinate-dependent O2 uptake in permeabilized cells follows the classical pathway. Oxidation of NADH by a membrane-rich fraction produced H2O2 as the end product and was insensitive to respiratory chain inhibitors. The presence of NADH-fumarate reductase was demonstrated in membrane-rich fraction and fumarate could reduce H2O2 production from NADH indicating fumarate to be an endogenous substrate for accepting electrons from NADH. A differential route for NADH oxidation was further confirmed by NADH cytochrome c reductase insensitivity to antimycin. A tentative scheme for electron transfer pathway in this organism is proposed in which a reversal of Krebs cycle enzymes occur producing succinate that can be excreted or oxidized depending upon the energy demands of the cell. Inhibition studies also suggest bifurcations of the respiratory chain that can be of minor importance for the organism. PMID- 8720176 TI - A group of Giardia lamblia variant-specific surface protein (VSP) genes with nearly identical 5' regions. AB - The surfaces of Giardia lamblia trophozoites contain one of a set of variant specific surface proteins. The genes encoding these proteins are highly conserved at the 3' terminus, but frequently demonstrate little similarity in the remainder of the coding region. This report describes a family of vsp genes highly similar to a repeat-containing vsp gene (vspC5) at the 5' coding and flanking regions, but which diverge abruptly from vspC5 in the first repeat and do not themselves contain full copies of the repeat. This observation suggests the possibility that recombination among different vsp genes may have played a role in development of the vsp gene repertoire. PMID- 8720175 TI - Effects of 3' untranslated and intergenic regions on gene expression in Trypanosoma cruzi. AB - The effects of 3' untranslated regions (UTR) and intergenic regions (INT), from various Trypanosoma cruzi stage-specific and constitutive genes, on the expression of the reporter firefly luciferase gene (luc), were studied using stable episomal transformation. The 3' UTR influenced luciferase expression by changing the steady-state level and/or the translation efficiency of luc mRNA. Glycoprotein 72 gene (gp72), glycoprotein 85 gene (gp85) or amastin gene (ama) 3' UTR decreased the luc mRNA level 6- to 14-fold, compared to the glyceraldehyde 1 phosphate dehydrogenase gene (gapdh) 3' UTR, in epimastigotes. Luciferase activity decreased in parallel with the luc mRNA level in transformants utilizing the gp85 or ama 3' UTR, whereas luc mRNA containing the gp72 3' UTR showed approximately 5-fold higher translation efficiency than luc mRNA containing a minimal 3' UTR. In amastigotes, the inhibitory effect of the ama 3' UTR observed in other life cycle stages was abolished and luciferase expression was stimulated 16-fold. The overall stage-specific difference mediated by the ama 3' UTR, between epimastigotes and amastigotes, was approximately 100-fold. INT, which was expected to influence polyadenylation efficiency, of gapdh, gp72, or heat shock protein 60 gene inserted after gapdh 3' UTR increased luc mRNA 2- to 8-fold, whereas gp85 INT slightly decreased luc mRNA. By separating effects attributable to the 3' UTR and INT, this study shows the effects of 3' UTR on RNA levels and translational efficiency in T. cruzi. PMID- 8720177 TI - Sporozoites of Toxoplasma gondii lack dense-granule protein GRA3 and form a unique parasitophorous vacuole. AB - The invasion of host cells by sporozoites of Toxoplasma gondii leads to the formation of parasitophorous vacuoles that are distinctly different from those surrounding tachyzoites. In sporozoite-infected cells, the fluid-filled space surrounding the sporozoite is many times larger in volume than the sporozoite, essentially lacks granular or tubular structures, and has no detectable continuous parasitophorous vacuolar membrane when prepared by conventional electron microscopic methods. Consistent with the ultrastructural differences, dense-granule protein GRA3, which associates with the parasitophorous vacuolar membrane of tachyzoites, was not detected by indirect immunofluorescence in sporozoite-infected cells 2-12 h post-inoculation or by Western blot analysis of sporozoite extracts. Western blots incubated with the alpha ROP/DG antiserum, which recognizes tachyzoite rhoptry and dense-granule proteins, revealed numerous other antigenic differences between sporozoites and tachyzoites. Cell cultures inoculated with sporozoites were monitored at various intervals for the expression of GRA3 and the developmentally-regulated tachyzoite surface protein SAG1. Expression of SAG1 and GRA3 was first observed in 30% of the sporozoite infected cells at 12 and 15 h post-inoculation, respectively, and in all intracellular parasites at 24 h. Parasite replication was only observed in sporozoite-infected cells that were positive for GRA3 and SAG1. Thus, these data indicate that sporozoites and their interaction with host cells differ substantially from tachyzoites and the expression of tachyzoite-specific proteins is likely required for parasite replication. PMID- 8720178 TI - Pyruvate dehydrogenase complex from the primitive insect trypanosomatid, Crithidia fasciculata: dihydrolipoyl dehydrogenase-binding protein has multiple lipoyl domains. AB - The pyruvate dehydrogenase complex (PDC) has been purified to apparent homogeneity from the insect trypanosomatid, Crithidia fasciculata, a member of the most primitive eukaryotic group to contain mitochondria. Separation of the purified PDC by SDS-PAGE yielded five bands of 70 (p70), 60 (p60), 55, 46 and 36.5 kDa, which appeared to correspond to dihydrolipoyl dehydrogenase binding protein (E3BP), dihydrolipoyl transacetylase (E2), E3, E1 alpha and E1 beta, respectively. The purified complex did not exhibit endogenous PDHa kinase activity. p70 was much less abundant than p60. Polyclonal antisera raised against p70 did not cross-react with p60, and antisera raised against p60 did not cross react with p70, suggesting that p60 did not arise from p70 by proteolysis. Both p70 and p60 contained similar amino terminal sequences. Both sequences contained the MPALSP motif similar to sequences present in both E3BP and E2 from other sources. Incubation of the purified PDC with [2-14C]pyruvate in the absence of CoA resulted in the acetylation of both p70 and p60, suggesting that both proteins contained lipoyl domains, but the specific incorporation of label into p70 was significantly greater than for p60. Limited proteolysis of the acetylated complex with trypsin yielded two major fragments derived from p60 of 35 and 30 kDa, corresponding to E2L and E2I, and one major acetylated fragment of 58 kDa derived from p70. Therefore, these results suggest that p70 is an E3BP and given its apparent M(r) and degree of acetylation, it contains multiple lipoyl domains. PMID- 8720179 TI - Characterization of a Schistosoma mansoni cDNA encoding a B-like cyclophilin and its expression in Escherichia coli. AB - A cDNA encoding a Schistosoma mansoni cyclophilin (SmCyP) has been cloned by polymerase chain reaction amplification using degenerate oligonucleotides based on known conserved cyclophilin (CyP) sequences and by screening an expression cDNA library. The cDNA sequence encodes a 21.5-kDa protein, which shares 59% sequence identity with human CyP B. The SmCyP protein was expressed in Escherichia coli with a hexahistidine affinity tag at its amino terminus and antibodies to the purified (His6)-SmCyP fusion protein were raised in a rabbit. Fractionation of parasite material followed by immunoblot analysis revealed that schistosome CyP is a soluble protein. The N-terminus of the predicted protein contains a hydrophobic region, suggestive of a signal sequence. Accordingly, a recombinant SmCyP protein, lacking the first 23 amino acids was found to share the same gel electrophoretic mobility as the parasite-derived CyP protein, suggesting cleavage of a leader sequence. Hybridization of genomic DNA to a full length cDNA probe indicates that the SmCyP gene is present as a single copy. Immunohistological experiments in conjunction with confocal scanning laser microscopy and immune electron microscopy show that SmCyP is present in abundance in the adult worm as well as in the schistosomula. The function of CyP in the schistosome is presently unclear, but since its ligand, cyclosporin A, has antischistosomal activity, its function is expected to be a vital one. PMID- 8720180 TI - Cloning and characterization of two genes encoding Schistosoma mansoni elastase. PMID- 8720181 TI - Cloning of two actin genes from Schistosoma mansoni. PMID- 8720182 TI - A putative RNA helicase of the DEAD box family from Trypanosoma brucei. PMID- 8720183 TI - Addendum to "Trypanosoma brucei dihydrofolate reductase-thymidylate synthase: gene isolation and expression and characterization of the enzyme" [Mol. Biochem. Parasitol. 72 (1995) 11-22]. PMID- 8720184 TI - Erratum to "The construction of spliced leader cDNA libraries from the filarial nematode Brugia pahangi" [Mol. Biochem. Parasitol. 70 (1995) 241-245]. PMID- 8720185 TI - The need for longitudinal research. PMID- 8720186 TI - Changing the mind-set of nurses. PMID- 8720187 TI - Alternatives to traditional health care. PMID- 8720188 TI - Annual update of how each state stands on legislative issues affecting advanced nursing practice. AB - The change from a free-for-service system to a system run by managed-care organizations has caused dramatic shifts in where and how health care is delivered. Within the managed-care systems, administrators need to show a good bottom line. Rapid restructuring of the system and the need for profit create many potential consequences (e.g., APNs squeezed out of participation on some provider panels; previously unattractive poor patients are now attractive because they can provide an ongoing revenue base; the use of more unlicensed personnel to provide patient services). Private businesses and the federal government like managed-care health care systems as an answer to the out-of-control escalating costs of health care. Managed care is likely to stay with us for a long while. Because political force comes from money and there is plenty of that in health care insurance corporations, managed care is likely to be with us for a long time. Regulations and control of these managed-care systems will probably be difficult. However, though big business has the money, APNs have the feet, hands and patience to seek further regulation of these huge corporations (e.g., antidiscrimination language to prevent exclusion from provider panels). With the reality of managed care, MDs are no longer in control of the health care system. The AMA's recent attempt to control other providers is tantamount to one ship officer attempting to control another on the Titanic's (i.e., fee-for-service system's) last evening. In a few states, fee-for-service is still predominant, but the majority of states are rapidly moving into megaconglomerate, finance driven managed care run by business administrators. Things are getting so complex in today's world of corporate takeovers, managed-care megaconglomerates, and power-brokering politics that it is easy to understand why millions of Americans drown themselves in relatively simplistic good-versus-evil entertainment dramas. Huge changes in how the system operates have happened this year while the nation was busy following the Simpson trial. But NPs and other APNs must be very vigilant. We must ensure a place in this new system. Our patients and the nation need us! PMID- 8720189 TI - Guidelines for developing a nurse practitioner practice. AB - Nurse practitioners must take an active role in defining and establishing their practices. In many cases, medical and administrative staffs develop the NP job description. As a result, many NPs are finding themselves in unsatisfactory, often task-oriented roles. This problem is largely due to NPs' inexperience in establishing practices and a lack of documentation of current practices. When developing the NP practice within the National Cancer Institute-Medicine Branch, it was apparent that the nursing literature offered no information on how to establish an NP practice. This article presents guidelines for developing an NP practice and is based on information obtained during the developmental stages of the nurse practitioner practice at the National Cancer Institute-Medicine Branch. PMID- 8720190 TI - Evaluation of certain food additives and contaminants. AB - This report presents the conclusions of a Joint FAO/WHO Expert Committee convened to evaluate the safety of various food additives and contaminants, with a view to recommending acceptable daily intakes for humans, and to prepare specifications for the identity and purity of food additives. The first part of the report contains a general discussion of principles governing the toxicological evaluation of food additives and contaminants, including flavouring agents, and the establishment and revision of specifications, with comments concerning limits for arsenic, lead and heavy metals. A summary follows of the Committee's evaluations of toxicological data on various specific food additives (including tert-butylhydroquinone, mineral oils, waxes and alitame) and on beta cyclodextrin, nitrate, nitrite, potassium bromate and the contaminants ochratoxin A and patulin. Annexed to the report are a table summarizing the committee's recommendations for acceptable daily intakes of these substances and changes in the status of specifications, and a list of further toxicological studies required or desired. PMID- 8720191 TI - Use of phenotypic and genotypic fingerprinting methods in the strain identification of Aspergillus fumigatus. AB - Aspergillus fumigatus isolates (n = 6) from a lung transplant recipient, one isolate from a patient who had been on the same ward and a reference strain (NCPF 2140) were compared using three typing methods: SDS-PAGE, immunoblotting with serum from the transplant patient and random amplified polymorphic DNA (RAPD) assay. Neither the SDS-PAGE, immunoblot nor RAPD assay with single primers revealed differences between the eight isolates. Digestion of one primer product with the endonuclease EcoRI discriminated between the six patient isolates and the other two strains. The RAPD assay using pairwise combined primers showed identical patterns for the patient's strains but differentiated between the two other strains. It is concluded that any single technique may fail to detect strain differences and that a spectrum of typing methods is necessary in order to reveal or to exclude cross-infections with Aspergillus fumigatus. PMID- 8720192 TI - Post-antibiotic effect and post-expositional polyene antagonism of azole antifungal agents in Candida albicans: dependence on substance lipophilia. AB - The lipophilic azoles itraconazole (ICZ), ketoconazole (KCZ) and miconazole (MCZ) have two things in common regarding their effect on Candida albicans. First, these azoles cause a growth inhibition that persists for at least 24 h after exposure (post-antibiotic effect), although this is only occasionally observed for ICZ. Secondly, these substances cause a decrease in the fungicidal activity of amphotericin B (AMB, 1 mg l-1) upon subsequent exposure to this drug. In contrast, fluconazole (FCZ) exhibits neither of these two effects. Further tests suggest that both of these phenomena observed may be related to the non-covalent binding of the three lipophilic azoles to lipophilic cytoplasmic components of yeast cells. With fluconazole, such bonds seem to be much weaker. The amount of relatively hydrophilic fluconazole that is bound non-specifically to the fungal cell is evidently too low to produce long-lasting post-exposure effects like those caused by lipophilic azoles. PMID- 8720193 TI - Testing susceptibility of Candida species to fluconazole and itraconazole using the microdilution assay. AB - A microdilution system was established for testing the susceptibility of Candida species to fluconazole and itraconazole. The assay used a sodium phosphate buffered (0.1 mol l-1) Casitone/glucose medium (pH 7.2) supplemented with potassium, iron, magnesium, trace elements and vitamins. Tests were read photometrically after 24 h, and the minimum inhibitory concentration (MIC) was defined as the IC90. In nearly all strains sharp end points were observed. The MICs against Candida species without any known pre-exposure to azoles were found to range from 0.2 to 1.56 micrograms ml-1 for fluconazole and from 2.3 to 12 ng ml-1 for intraconazole. For strains from candidosis patients refractory to treatment with fluconazole the MICs of fluconazole ranged from 6.25 to 100 micrograms ml-1, while those for itraconazole varied between 12 and > 450 ng ml 1. The strains did not respond congruent to both azoles. A similar disparity of the MICs was observed with Candida tropicalis and Candida krusei. The unusually low MICs of itraconazole were attained because (1) the dilution series was prepared from a preformed (concentrated) dilution series in 75% dimethylsulphoxide and not directly by serial dilution in the broth and (2) the incubation was performed in microtitre plates made of quartz glass rather than in the generally used polystyrene microtitre plates. PMID- 8720194 TI - Itraconazole and flucytosine+itraconazole combination in the treatment of experimental cryptococcosis in hamsters. AB - The efficacy of two different daily doses of itraconazole (ITRA) and the combination of flucytosine (5-FC) with ITRA in the treatment of an experimental model of cryptococcosis in hamsters was studied. Five groups of 20 animals each were inoculated by the intracardiac route with 10(5) cells of Cryptococcus neoformans. Treatment started 3 days after the infection, and was administered by gavage for 30 days. ITRA was applied at a daily dose of 25 mg kg-1 or 50 mg kg-1 and the combination of 5-FC and ITRA was given at 75 mg kg-1 day-1 or 50 mg kg-1 day-1 respectively. One group of 20 hamsters received the vehicle and was used as a control group. Treatment evaluation was based on the following parameters: number of surviving animals 60 days after the infection; presence of encapsulated yeasts on microscopic examination of wet preparations of brain, lungs, liver, spleen and kidneys at necropsy; and brain qualitative (massive seeding) and quantitative cultures (determination of colony forming units, CFU). ITRA 50 (50 mg kg day-1) was the most effective treatment according to the studied parameters; 70% of brain cultures became negative and 95% of the treated hamsters survived to the end of the study period. ITRA efficacy was dose dependent. The combination of ITRA with 5-FC was less effective than administering the drugs separately; the reason for this finding is not known. The results obtained in this study should encourage the use of high doses of ITRA in cases of disseminated cryptococcosis in humans. PMID- 8720195 TI - Specific and non-specific parameters of the host defence system in patients with superficial fungal infections. AB - Some measures of systemic host defences, i.e. white cell counts, lymphocyte subsets, delayed-type hypersensitivity and polymorphonuclear leucocyte functions, were evaluated in 42 patients suffering from cutaneous superficial mycoses and in 35 healthy volunteers. Patients were divided according to the extent of their lesions into two groups: group A (30 patients) with skin involvement > or = 30% and group B (12 patients) with skin involvement < or = 1%. No significant abnormalities in systemic defence mechanisms were observed in group B patients, whereas multiple alterations of polymorphonuclear phagocyte activities, i.e. chemotaxis, phagocytosis, mitogen-induced superoxide anion production, were observed in patients with extensive lesions. The values of these phagocytic functions remained significantly reduced (P < 0.01) in these patients after recovery and during a 6-month follow-up period. PMID- 8720196 TI - Infusion-related toxicity of three different amphotericin B formulations and its relation to cytokine plasma levels. AB - A prospective study was performed to compare the infusion-associated toxicity of three different amphotericin B preparations and to correlate acute side-effects with plasma levels of tumour necrosis factor-alpha (TNF-alpha), interleukin-6 (IL 6) and interleukin-1 receptor antagonist (IL-1-RA) during and after the infusions. Six adult neutropenic patients with acute leukaemia suffering from suspected or documented systemic fungal infections were treated on three consecutive days with conventional amphotericin B (AmB), liposomal AmB (AmBisome) and AmB mixed in lipid emulsion (AmB/lipid). Drugs were given over 1-2 h. Drug induced toxicity was monitored every 30 min for 4 h. Plasma levels of the three cytokines were determined using commercially available enzyme-linked immunosorbent assay (ELISA) techniques. Four of six patients showed toxicity after AmB and AmB/lipid infusions; only one patient reacted to liposomal AmB. Clinical toxicity was associated with increases in TNF-alpha plasma levels during two of four infusions of AmB and three of four infusions of AmB/lipid. Major increases in IL-6 occurred during three of four infusions of AmB and during all four AmB/lipid infusions associated with clinical toxicity. Three of four AmB infusions and all four AmB/lipid infusions accompanied by clinical toxicity were associated with major increases in IL-1-RA plasma concentrations. Liposomal AmB was better tolerated than AmB and AmB/lipid. This formulation also caused the lowest liberation of all three cytokines tested. The severity of clinical symptoms did not correlate closely with absolute cytokine plasma levels. The findings provide further evidence that expression of TNF-alpha, IL-6 and IL-1-RA plays an important role in mediating AmB-related acute toxicity in vivo. PMID- 8720197 TI - Peracute disseminated fatal Aspergillus fumigatus sepsis as a complication of corticoid-treated systemic lupus erythematosus. AB - Immunocompromised and granulocytopenic patients and those receiving long-term or high-dose corticoid treatment are predisposed to disseminating Aspergillus infections. However, Aspergillus infection has been described only rarely in patients with autoimmune diseases. We report on a woman suffering from systemic lupus erythematosus treated by antibiotics and high-dose corticosteroids, a primary risk factor, who developed a peracute disseminated fatal Aspergillus fumigatus infection involving the central nervous system. The present case is compared with 10 previous reports of invasive aspergillosis in systemic lupus erythematosus found by a literature search. PMID- 8720198 TI - Cerebral Pseudallescheria mycosis after near-drowning. AB - Scedosporium apiospermum, an anamorph of Pseudallescheria boydii, is a hyalohyphomycete which can be mistaken, histologically, for Aspergillus. S. apiospermum is clinically distinguished by resistance to most antifungals and its ability to cause invasive mycoses in immunocompetent patients. Here we present a typical case of lethal cerebral Pseudallescheria mycosis in a child after near drowning. PMID- 8720199 TI - Macroscopic and microscopic characteristics of an African Blastomyces dermatitidis strain. AB - This report describes the macro- and microscopic characteristics of a strain of Blastomyces dermatitidis isolated in an Italian hospital from a Libyan patient suffering from cutaneous blastomycosis. The fungus was isolated with great difficulty because of the presence of a Proteus species on the lesion. Conversion of the mycelial into the yeast-like phase achieved the best results, using Columbia ANC culture medium. After 5-6 days, this transformation was only partial and presented swollen hyphae and yeast-like cells. The biological characteristics were typical of B. dermatitidis strains of African origin. The differences between African and North American strains are confirmed, in agreement with findings already reported in the literature. PMID- 8720200 TI - An atypical isolate of Paracoccidioides brasiliensis. AB - An atypical strain of Paracoccidioides brasiliensis was obtained from a patient with disseminated paracoccidioidomycosis. This isolate, which has been maintained in the laboratory for 4 years by successive subcultures (30-40 days) in Fava Netto medium, produces cerebriform colonies characteristic of the yeast phase of the fungus when left at room temperature. Microscopy shows a predominance of yeasts with multiple buds, side by side with some branching filamentous structures presenting intercalary chlamydospores. The strain does not develop well at 37 degrees C, suggesting heat sensitivity. Storage at 4 degrees C does not change its macro- or microscopic characteristics. These findings support the hypothesis raised by others that the dimorphism of P. brasiliensis may not be exclusively heat dependent. Studies are currently being developed in our laboratory to better characterize this isolate in terms of biochemical peculiarities, degree of virulence and susceptibility/resistance to antifungal drugs. PMID- 8720201 TI - Fluconazole in the therapy of tropical deep mycoses. AB - A clinical study was conducted to test the efficacy of fluconazole in the treatment of tropical deep mycoses. Two out of four patients with zygomycosis due to Conidiobolus coronatus who were treated with the drug were completely cured; the other two patients exhibited considerable improvement but could not be followed up. Two patients with eumycetoma, one due to an Acremonium sp. and one due to Pseudallescheria boydii, were treated successfully, whereas another patient with a eumycetoma caused by an unidentified fungus could not be followed up. A complete cure was achieved with one patient with African histoplasmosis and one with candiduria. A case of cerebral phaeohyphomycosis due to Cladosporium sp. showed some improvement but the patient later developed meningitis and died. PMID- 8720202 TI - Cell-surface hydrophobicity and lipolysis as essential factors in human tinea nigra. AB - Hydrophobic adhesion of cells of the black yeast Hortaea werneckii (Horta) Nishimura & Miyaji, causative agent of human tinea nigra, was established by microbial adhesion to hexadecane (MATH) and adhesion to polystyrene, and compared with adhesion of other species of black yeasts. Additional ecophysiological tests were performed. Hortaea werneckii cells proved to have a high degree of hydrophobicity (98.5% MATH). The species is unable to degrade keratin, but shows significant lipolytic activity. It is concluded that H. werneckii is a commensal, that shows lipophilic adhesion to human skin and survives by the assimilation of excretion products. PMID- 8720203 TI - Tinea pedis in members of the Japanese Self-defence Forces: relationships of its prevalence and its severity with length of military service and width of interdigital spaces. AB - We examined the relationship between prevalence and severity of tinea pedis and the length of service and the width of the spaces between the toes in 74 members of the Japanese Self-Defence Forces (SDF) undergoing special training. The subjects were divided according to the width of these spaces into: group I, wide; group II, fairly wide; and group III, closed. The severity of tinea pedis was determined by its duration and the extent of the lesions in the 49 subjects who had tinea pedis. The combined prevalence of tinea pedis and tinea unguium was 66%. There was a tendency for the prevalence to be higher in subjects who had served for 10 years or more in the SDF than in those with fewer than 10 years of service. Classified by the disposition of their toes, 10 subjects fell into group I, 34 into group II, and 30 into group III. The prevalence of 90% (27/30) in group III was significantly higher than in the other groups. A significant positive correlation was seen between length of SDF service and severity. Subjects with both a long service record and closed interdigital spaces showed both a high prevalence and marked severity. PMID- 8720204 TI - Croconazole in the treatment of tinea pedis. AB - Croconazole hydrochloride is a new antifungal agent of the imidazole type. A 1% croconazole cream (Pilzcin, Merz + Co., D-Frankfurt/Main) was investigated in a multicentre trial involving 132 patients (mean age 46.7 +/- 15.5 years; 69 men, 63 women) suffering from tinea pedis (interdigital space, n = 86; other foot sites, n = 46). The fungal infections were caused predominantly by Trichophyton rubrum (interdigital space, n = 43; other foot sites, n = 33), followed by other Trichophyton and Candida species. The patients were treated once daily for a period of up to 3 weeks. In the majority of cases complete cure was achieved at both locations. On fungal microscopy no fungi were seen after 3 weeks in 82.6% of patients in the 'interdigital space' group and in 80.4% of patients in the 'other foot sites' group. Two weeks after the end of the treatment no fungi could be found in culture in 81.4% of the patients in the 'interdigital space' group and in 80.4% of patients in the other group. All skin symptoms of the mycoses (itching, scaling, erosions, reddening) decreased during the observation period. In 82.6% of patients in the 'interdigital space' group and in 76.1% of patients treated in the 'other foot sites' group efficacy was rated by the physician as good or very good. Tolerability was evaluated as good or very good in 90.7% and 89.1% of the cases. Local skin intolerabilities were not observed. The therapeutic results show that croconazole cream, when applied once a day, is an effective and well-tolerated drug for the treatment of tinea pedis, regardless of its location. PMID- 8720205 TI - Fungal contamination of the floors of swimming pools, particularly subtropical swimming paradises. AB - We compared the level of dermatophyte contamination of the floors of traditional swimming pools and of subtropical swimming paradises, a new type of indoor swimming pool introduced in Belgium in 1981. From the results of routine hygiene inspections over several years and of a comparative study, we can conclude that the floors of subtropical swimming pools are much more highly contaminated by dermatophytes than the floors of traditional swimming pools. This higher degree of contamination is due to the huge number of visitors, the complexity of construction, the choice of materials and the long opening hours. In both types of swimming pools, the predominant species was Trichophyton mentagrophytes var. interdigitale, followed by T. rubrum. Other dermatophytes, such as Microsporum canis and M. gypseum, were isolated sporadically. PMID- 8720206 TI - Radiofrequency ablation of atrial insertion of left-sided accessory pathways guided by the "W Sign". AB - INTRODUCTION: The aim of this study was to evaluate the efficacy of radiofrequency (RF) ablation of the atrial insertion of left-sided accessory pathways with guidance by a specific morphologic characteristic of the local electrogram, which we call the "W sign." This represents the shortest local atrioventricular (AV) interval during sinus rhythm in patients with manifest preexcitation or the shortest local VA interval during AV reciprocating tachycardia and/or ventricular pacing in patients with concealed accessory pathways. METHODS AND RESULTS: The transseptal technique was used in 31 patients (18 men, 13 women; aged 32 +/- 13 years), and RF ablation of 33 accessory pathways (26 manifest and 7 concealed) was attempted. Patients presented with palpitations (n = 16), presyncope (n = 10), or syncope (n = 5). The clinical arrhythmia was AV reciprocating tachycardia (n = 24) or atrial fibrillation (n = 7). In 21 patients (68%) electrophysiologic study and RF ablation were performed at a single session. Accessory pathways were left posteroseptal (n = 5) or left free wall (n = 28). The "W sign," formed from merging of the local atrial and ventricular electrograms, was identified at all successful sites prior to ablation. Ablation was successful in all patients. A median of 7 RF lesions were delivered per patient. The fluoroscopy time was 76 +/- 48 minutes; total procedure time was 5.4 +/- 1.9 hours. No significant complications occurred. Early recurrence (< or = 24 hours) occurred in 1 patient; during 6 +/- 4 months, accessory pathway conduction recurred in another patient. CONCLUSION: We conclude that RF ablation of the atrial insertion of left accessory pathways can be very successful when guided by the "W sign." PMID- 8720207 TI - Detailed mapping for catheter ablation of left-sided accessory pathways: search for the "magic" signals from A to V and W to Z. PMID- 8720208 TI - Stability: an ICD detection criterion for discriminating atrial fibrillation from ventricular tachycardia. AB - INTRODUCTION: The purpose of this study was to review a new implantable cardioverter defibrillator (ICD) detection criterion, stability, to determine if it can effectively discriminate rapid rhythms of atrial fibrillation from ventricular tachycardia. Inappropriate shocks for rapid atrial fibrillation limit the acceptance of ICDs. The advent of an additional detection criterion, stability, has been postulated to be of value in discriminating rapid atrial fibrillation, which may not warrant treatment, from ventricular tachycardia, which obviously does warrant therapy deliver. METHODS AND RESULTS: Twenty-six patients were studied during 32 episodes of rapid atrial fibrillation and 24 episodes of monomorphic ventricular tachycardia below 220 beats/min. Each rhythm was repeatedly evaluated by the device at each of the seven stability values available (8, 16, 23, 31, 39, 47, and 55 msec) and then classified as stable or unstable. Upon completion of this acute study, 32 ICD patients had the stability feature activated and were followed for proper arrhythmia treatment by the device. Using stability windows from 8 to 47 msec, all atrial fibrillation rhythms were appropriately classified as unstable. Three of 6 were classified correctly for the 55-msec window. All ventricular tachycardia rhythms were appropriately classified as stable from all stability windows from 8 to 55 msec. Clinical follow-up confirmed appropriate therapy delivery when coupled with sustained rate duration (SRD). Thirty-two patients followed for 292 patient months had no episodes of untreated ventricular tachycardia with 428 successfully classified as stable and treated. Only three episodes of suspected atrial fibrillation resulted in therapy delivery as the rhythm duration exceeded the SRD of 30 seconds. CONCLUSIONS: The CPI Ventak PRx ICD is highly reliable in appropriately classifying atrial fibrillation as unstable and monomorphic ventricular tachycardia as stable for most stability windows evaluation tachycardias below 220 beats/min. As a result, when testing of atrial fibrillation is not possible, we recommend the routing programming of this stability feature at the 31-msec window with an SRD of 30 seconds. The reliability of this device in discriminating atrial fibrillation from monomorphic ventricular tachycardia may have important clinical implications for other tiered therapy ICDs with this feature as well as for future ICDs in development. PMID- 8720209 TI - Radiofrequency delivery through a cooled catheter tip allows the creation of larger endomyocardial lesions in the ovine heart. AB - INTRODUCTION: The delivery of radiofrequency (RF) energy through conventional catheter electrodes is often associated with coagulation necrosis at the tissue electrode interface, with resultant impedance rise and limited lesion size. This study was performed to examine the effects of catheter tip cooling during RF delivery, to test the hypothesis that such cooling would decrease the likelihood if impedance rise and allow the creation of larger endomyocardial lesions. METHODS AND RESULTS: The experiments were performed in eight open chest, anesthetized sheep. RF lesions were created within both ventricular chambers of each animal through a catheter tip that could be cooled with a saline perfusate. Assignment of cooled versus noncooled RF delivery to either ventricle was alternated from one animal to the next. In each set of experiments, lesion volumes relative to the mode of RF delivery were compared. The mean power delivered via the cooled electrode (22.04 +/- 4.51 W) was significantly higher than that delivered via the noncooled electrode (6.10 +/- 2.47 W; P < 0.001). The mean duration of RF delivery was 42.7 +/- 11.2 sec for noncooled lesions versus 49.2 +/- 6.8 sec for cooled lesions (P < 0.01). Mean lesion volume was 436.07 +/- 177.00 mm3 for noncooled RF delivery versus 1247.78 +/- 520.51 mm3 for cooled RF delivery (P < 0.001). This significantly larger lesion size with cooled RF delivery was associated with no instance of impedance rise in 27 attempts versus 11 impedance rises in 28 attempts with noncooled RF (P < 0.001). CONCLUSIONS: Delivery of RF energy through a cooled catheter tip allows the creation of larger endomyocardial lesions by limiting the occurrence of impedance rise despite the delivery of greater energy. These observations suggest that, under certain conditions, resistive tissue heating at a distance from the site of current delivery may play an important role in RF ablation therapy. PMID- 8720210 TI - A prospective randomized comparison in humans of 90-mu F and 120-mu F biphasic pulse defibrillation using a unipolar defibrillation system. AB - INTRODUCTION: Capacitance is known to influence defibrillation. Optimal biphasic waveform capacitance for transvenous unipolar defibrillation systems in man is currently being defined. In an effort to improve defibrillation efficacy, we examined the relative defibrillation efficacy of a 65% tilt biphasic pulse from a 90-mu F capacitor compared to a 65% tilt biphasic pulse from a 120-mu F capacitor in a prospective, randomized fashion in 16 consecutive cardiac arrest survivors undergoing defibrillator surgery. METHODS AND RESULTS: The transvenous unipolar pectoral defibrillation system uses a single endocardial RV anodal defibrillation coil and the shell of an 80-cc volume (88 cm2 surface area) pulse generator (Medtronic Model 7219C PCD "active CAN") as the cathode for the first phase of the biphasic shock: RV+ --> CAN-. Defibrillation thresholds for each capacitance were determined prospectively in a randomized fashion. The defibrillation threshold results for the 90-mu F capacitance were: leading edge voltage 383 +/- 132 V; stored energy 7.4 +/- 5.0 J; and resistance 57 +/- 10 omega. The results for the 120-mu F capacitance were: leading edge voltage 315 +/- 93 V (P = 0.002); stored energy 6.5 +/- 3.7 J (P = 0.21); and resistance 57.0 +/- 11 omega (P = 0.87). CONCLUSIONS: We conclude that 90-mu F, 65% tilt biphasic pulses used with unipolar pectoral defibrillation systems have equivalent stored energy defibrillation efficacy compared to 120-mu F, 65% tilt pulses. Use of lower capacitance is possible in present implantable defibrillators without compromising defibrillation. PMID- 8720211 TI - Failure of impulse propagation in a mathematically simulated ischemic border zone: influence of direction of propagation and cell-to-cell electrical coupling. AB - INTRODUCTION: It is suggested that heterogeneous extracellular potassium concentration, cell-to-cell coupling, and geometric nonuniformities of the ischemic border zone contribute to the incidence of unidirectional block and subsequent development of lethal ventricular arrhythmias. METHOD AND RESULTS: A discrete electrical network was used to model a single cardiac fiber with a [K+]e gradient characteristic of an ischemic border zone. Directional differences in propagation were evaluated by creating discrete regions with increased gap junctional resistance within the [K+]e gradient. Furthermore, the effect of homogeneity/heterogeneity of call length on impulse propagation through the [K+]e gradient in the presence of increased gap junctional resistance was evaluated. The results indicate that failure of impulse propagation occurs at the junction between partially uncoupled and normally coupled cells. Furthermore, propagation failure was more likely to occur as the impulse propagated from a region of high [K+]e to low [K+]e. Heterogeneity in cell length contributes to the variability in the occurrence of unidirectional and bidirectional block. CONCLUSIONS: The onset of cellular uncoupling in an ischemic border zone may interact with the inherent [K+]e gradient leading to unidirectional conduction block. This mechanism may be important for the generation of reentrant arrhythmias at the ischemic border zone. PMID- 8720212 TI - Radiofrequency ablation of idiopathic left anterior fascicular tachycardia. AB - INTRODUCTION: A 45-year-old man with idiopathic ventricular tachycardia (VT) having a right bundle branch block configuration with right-axis deviation underwent an electrophysiologic test. METHODS AND RESULTS: Mapping demonstrated a site on the anterobasal wall of the left ventricle where there was an excellent pace map and an endocardial activation time of -20 msec, but radiofrequency catheter ablation at this site was unsuccessful. At a nearby site, a presumed Purkinje potential preceded the by 30 msec during VT and sinus rhythm, and catheter ablation was effective despite a poor pace map and an endocardial ventricular activation time of zero. CONCLUSION: Idiopathic VT with a right bundle branch configuration and right-axis deviation may originate in the area of the left anterior fascicle. A potential presumed to represent a Purkinje potential may be more helpful than endocardial ventricular activation mapping or pace mapping in guiding ablation of this type of VT. PMID- 8720213 TI - Variant preexcitation syndrome: a true nodoventricular mahaim fiber or an accessory atrioventricular pathway with decremental properties? AB - INTRODUCTION: The differentiation between a nodoventricular fiber and an accessory atrioventricular (AV) pathway with long conduction times and decremental properties could be very difficult even at detained electrophysiologic study. METHODS AND RESULTS: A 20-year-old male with a history of a wide QRS tachycardia underwent electrophysiologic study. Baseline intervals were normal. There was evidence of dual AV pathways, and a sudden increase in AH interval was associated with the emergence of a delta wave. The atrio-delta interval showed a progressive prolongation. The preexcited QRS complex was typical of a posteroseptal pathway, and the earliest ventricular activation site was recorded at the posteroseptal region. Retrograde conduction was exclusively over the normal conduction system. During ventricular extrastimulation, a sudden increase in HA interval was associated with anterograde conduction over the accessory pathway. The intervals between the stimulus artifact and the onset of the delta wave during atrial pacing from two atrial sites (S-Delta) were compared with those between the retrograde atrial electrogram on the His channel and the onset of the delta wave during ventricular pacing (A2HB-Delta). When pacing from the proximal coronary sinus, the shortest S-Delta interval did become shorter than the longest A2HB-Delta interval (155 vs 170 msec). CONCLUSION: The finding that the S-Delta interval could become shorter than the A2HB-Delta interval provides strong evidence that this accessory pathway was not connected to the AV node but arose directly from the atrial tissue of the posteroseptal region. PMID- 8720214 TI - Inappropriate sinus tachycardia: evaluation and therapy. AB - Inappropriate sinus tachycardia is an ill-defined clinical syndrome characterized by an increased resting heart rate accompanied by an exaggerated response to exercise or stress. It is not associated with underlying structural heart disease. The mechanism may involve a primary abnormality of the sinus node demonstrating enhanced automaticity or, alternatively, a primary autonomic disturbance with increase sympathetic activity and enhanced sinus node beta adrenergic sensitivity. The diagnosis of inappropriate sinus tachycardia is one of exclusion. It is most common in young females, with a disproportionate number employed in the health care field. Autonomic and electrophysiologic testing may be required in selected individuals to clarify the mechanism and rule out sinus node reentry or right atrial tachycardia. Therapy of inappropriate sinus tachycardia is empiric. Pharmacologic approaches include beta blockers or verapamil. Radiofrequency catheter ablation of the superior portion of the sinus node shows promise as a useful alternative in patients with refractory symptoms. PMID- 8720215 TI - Mechanisms and therapy of complex arrhythmias in pediatric patients. AB - In recent years, the distinction between the diagnostic and therapeutic techniques used in the assessment and management of pediatric and adult patients with arrhythmias has gradually blurred. Nonetheless, arrhythmias in the pediatric patient are still often different from the adult patient in one of two important ways. First, a variety of arrhythmia mechanisms remain relatively unique to the pediatric population, some because of developmental issues and others because of early presentation of an incessant tachycardia. Second, the presentation and management of certain arrhythmias is sometimes markedly affected by patient age or the presence of structural congenital heart disease. A sampling from each of the above categories is reviewed and discussed. PMID- 8720216 TI - What is the tachycardia mechanism?--II. PMID- 8720217 TI - Cesium and the pacemaker current. PMID- 8720218 TI - Watch out for reference inaccuracies. PMID- 8720219 TI - Authors disagree about probability estimates. PMID- 8720220 TI - Physicians' responses to patient vignettes: are they valid? PMID- 8720221 TI - The residency program's 25th anniversary year: a national survey. PMID- 8720222 TI - Primary care of lesbian and gay patients: educating ourselves and our students. AB - Although a significant proportion of the population is gay or lesbian, physicians receive little formal training about homosexuality, and the unique health care needs of these patients are often ignored. Gay men and women may have higher rates of depression, suicide, alcoholism, certain cancers, and cardiovascular disease than their heterosexual counterparts. In addition, they are at risk of being victims of violence because of their sexual orientation. Due to fear of stigmatization by the medical community, the most significant health risk for lesbians, gays, and bisexuals may be that they avoid routine health care. Gay youth are particularly vulnerable to internal and external pressures, resulting in higher rates of substance abuse, suicide, and homelessness. Older gay men and women, who generally view themselves positively, may be troubled by declining health and loneliness. Physicians can improve the health care of gay and bisexual men and women and their families by maintaining a non-homophobic attitude toward these patients, distinguishing sexual behavior from sexual identity, communicating with gender-neutral terms, and maintaining awareness of how their own attitudes affect clinical judgment. Medical educators should avoid making assumptions about the sexuality of their residents and students. Institutions need to realize that the presence of supportive heterosexual and openly gay faculty will help create an environment that fosters learning for all students. Scant research exists about the best ways to teach about the special challenges gay men and lesbians face. However, the majority of surveyed medical students prefer that issues regarding gays and lesbians be integrated throughout the entire medical school curriculum. PMID- 8720223 TI - Using audiotapes to monitor student-preceptor interactions in a decentralized clerkship. AB - BACKGROUND: Community-based preceptors are increasingly being used to provide clinical experiences for medical students; therefore, the issue of when and how to monitor the appropriateness of teacher-learner interactions in the clinical setting has become increasingly complex and urgent. This paper describes how an audiotaping program in a department of family medicine was designed to account for preceptor and student needs and what this audiotaping experience revealed about the teaching strengths and weaknesses of full-time and volunteer faculty. METHODS: Audiotaped case discussions between third-year students and preceptors were evaluated using a structured instrument that reflected important responsibilities of clerkship preceptors. Reviews were compared using chi-square to evaluate for significant differences between university and community preceptors in overall quality of interaction with students and in specific deficiency areas. Ninety-five percent confidence intervals were used to evaluate for significant differences in frequency of occurrence among deficiencies. RESULTS: A total of 376 audiotaped case discussions were reviewed between July 1992 and June 1994. University preceptors had significantly more "superior" interactions with students than their community peers (P < .05). However, university and community preceptors had nonsignificant differences when specific deficiency areas were compared. Deficiencies in students who were allowed to present cases and assessments occurred significantly less frequently, while deficiencies in preceptors using open-ended questions occurred significantly more frequently than deficiencies in other areas. CONCLUSIONS: This study shows that volunteer faculty performed comparably to full-time university faculty in both the frequency of deficiencies in teacher-learner interactions identified and in the types of deficiencies identified. Both university and volunteer faculty had the most deficiencies in higher-order teaching skills--those involving synthesizing and assessing student knowledge and selecting appropriate questions based on that information. Audiotaped case discussions have a potentially significant role in monitoring teacher-student interactions, particularly on decentralized clinical rotations. PMID- 8720224 TI - Environmental health training: a survey of family practice residency program directors. AB - BACKGROUND: The Institute of Medicine and the American College of Physicians have advocated that physicians broaden their participation in the environmental aspects of medical care. Accordingly, both organizations recommend training of future primary care physicians for greater competency in and appreciation of this area of medicine. This study assessed the present emphasis on environmental health in family practice residency programs by examining the extent program directors expect graduates to have specific competencies in environmental medicine. METHODS: A written survey was mailed to directors of all 393 family practice residency programs listed in the 1993 Directory of Family Practice Residency Programs. Respondents were asked to indicate the extent to which each of eight environmental health competencies was expected of their graduates. The list of environmental health competencies was based on the literature and on interviews with family practice educators. Perceptions about the "present" and "ideal" environmental health emphasis in their programs were also measured. RESULTS: A total of 262 completed surveys were returned for a response rate of 67%. Respondents reported that they expected their graduates to have general knowledge of and competence in environmental health areas that pertain to patient care. Competencies with social and political implications were least likely to be expected. Two thirds of respondents indicated that "minimal emphasis" is presently placed on environmental health. Seventy percent indicated that the "ideal" amount of emphasis placed on this topic is "moderate." CONCLUSIONS: This survey's results suggest that family practice residency program directors expect their graduates to know basic environmental health concepts and be skilled in related aspects of patient care. The development of environmental health training programs must take into account that environmental health may be viewed as a topic of secondary importance and that in most residencies, faculty expertise in this area is limited. For this reason, a self-tutoring strategy, aimed toward the educational needs of both faculty and residents, is recommended. PMID- 8720225 TI - Medical students' and private family physicians' perceptions of family practice. AB - BACKGROUND: Many studies have attempted to determine what factors influence medical students' career selections. This study determined students' perceptions of family practice, examined what variables influenced these perceptions, and measured whether medical students' perceptions were similar to or different from those of practicing family physicians. METHODS: The two populations studied were students sampled at an Ohio medical school over the first 3 years of their training and a random sample of Ohio family physicians. Each subject was asked to complete a questionnaire that asked about perceptions of family practice and demographic information. RESULTS: A total of 719 (69%) students and 295 (59%) family physicians responded to the survey. Sixty-seven percent of the physician responders were actively involved in medical student teaching. More than 94% of the students had exposure to family physicians during their preclinical education. Students' perceptions were different than physicians' perceptions about lifestyle issues, patient care, and practice characteristics; over time, these approached the physicians' perceptions. However, the perceptions of family physicians and students about professional issues were similar. DISCUSSION: Medical students and family physicians have different perceptions about family practice. Students' perceptions become similar to physicians' perceptions as they progress through the educational system, while, at the same time, their interest in family practice declines. PMID- 8720226 TI - Family involvement in medical decision making. AB - BACKGROUND AND OBJECTIVES: The standard medical ethics model of decision making is based on the four principles of beneficence, nonmaleficence, autonomy, and justice. The recognized relationship is the physician and patient. This study considered the role(s) family plays in medical decision making. METHODS: Thirteen semi-structured interviews were conducted with a total of 39 patients and family members. Codebook and template analysis of the transcripts identified common themes. RESULTS: Patients and families identified three roles for family involvement in medical decision making--supporting the patient, being affected by the decision, and advocating for autonomy. CONCLUSIONS: Through these roles, patients acknowledge the context of family life in medical decision making, while families actively promote patient autonomy. Consideration of nonmedical burdens related to family roles and relationships takes an equal or higher priority than consideration of medical burdens. Family is, and should be treated as, a significant moral participant in medical decision making. PMID- 8720227 TI - Treatment of mild asthma with inhaled corticosteroids: is discontinuation of therapy possible? AB - BACKGROUND: This study investigated if long-term therapy with inhaled corticosteroids could be discontinued in mild asthma when patients are in a clinically stable phase of the disease. Data were derived from a 2-year randomized, controlled, bronchodilator intervention study in family practice. METHODS: The experimental (stop-steroid) group consisted of 19 asthmatic patients who had used inhaled corticosteroids daily during at least the year preceding this study and who stopped using these drugs because of participation in the bronchodilator intervention study. The control (no-steroid) group consisted of 70 patients with asthma who had not used corticosteroids in the year preceding the study. At the start of the study (8 weeks after stopping steroids), the two groups were completely comparable in all other relevant characteristics. During the 2-year study, patients were treated only with a bronchodilator (salbutamol or ipratropium bromide). Outcome measures were: exacerbations, symptoms, annual decline in forced expiratory volume in 1 second (FEV1), annual change in nonspecific bronchial responsiveness (PC20-histamine), and the need for additional corticosteroid therapy because of symptoms of increased airway obstruction. RESULTS: In the stop-steroid group, 12 of 19 patients (63%) dropped out during the study period because of a deterioration of their clinical condition and need for additional (inhaled) corticosteroid treatment. In the no steroid group, only eight patients dropped out for this reason (11%). In the stop steroid group, who did not use steroids for at least 1 year, the annual FEV1 decline was much larger than in the comparison subjects (165 vs 40 ml/yr). CONCLUSIONS: Stopping maintenance treatment with inhaled corticosteroids may not be advisable in all patients with mild asthma. Instead of stopping or interrupting treatment, family physicians are advised to determine the minimal effective daily dose of inhaled corticosteroids for each individual patient that provides adequate control of the disease. PMID- 8720228 TI - Ethnography: a stepwise approach for primary care researchers. AB - Ethnography is a research method borrowed from cultural anthropology and a disciplined method for understanding human conduct. It is a valid alternative to quantitative methods for primary care researchers. This article reviews the defining principles of ethnographic research, which uses principally observational methods to study actions and events that exist in natural situations. It is the job of ethnographic researchers to establish and frame their observations from the point of view of the people under observation. We present five steps fundamental to the development of an ethnographic research plan and a research case study that demonstrates the use of ethnography by a family physician. PMID- 8720229 TI - More on gay and lesbian people in our teaching environment. PMID- 8720230 TI - The original Glaxo Pathway Evaluation Program: a proposal for improvement. PMID- 8720231 TI - Effects of subject velocity on ground reaction force measurements and stance times in clinically normal horses at the walk and trot. AB - OBJECTIVE: The objective of the study reported here was to evaluate the effects of changing velocity on stance time and ground reaction force (GRF) measurements in horses at the walk and trot. DESIGN: Force plate gait analysis was used to evaluate clinically normal horses at variable velocities. Ground reaction force measurements and stance times were recorded and compared. ANIMALS: 12 adult horses. PROCEDURE: Data were obtained from 192 valid trials at the walk and 162 valid trials at the trot. Vertical, braking, and propulsive peak forces and impulses were measured. Pearson's correlation coefficients were determined for velocity and stance time and all measured forces and impulses in the forelimbs and hind limbs. Trials were divided into distinct velocity ranges. Trials obtained at velocities within the established ranges were analyzed to evaluate changes in vertical, braking, and propulsive peak forces and impulses at differing speeds within the walk and trot gaits. RESULTS: At the walk and trot, a significant negative correlation was found between velocity and forelimb and hind limb stance times. Velocity and stance time were significantly correlated with many of the GRF and impulse measurements. Velocity was significantly correlated with vertical and braking forces in the hind limbs at the walk, with vertical force in the forelimbs at the trot, and with braking force in the forelimbs and hind limbs at the trot. Velocity and stance time correlated significantly with forelimb and hind limb vertical impulses. Forelimb and hind limb stance times decreased significantly as velocity increased. Hind limb braking force increased and forelimb and hind limb vertical impulses decreased significantly as walk velocity increased. Forelimb braking force increased significantly between velocity ranges at the trot. CONCLUSIONS: Results of this study confirm that a significant negative linear correlation exists between subject velocity and stance times in clinically normal horses at the walk and trot. Significant correlations were also identified between velocity and many GRF measurements, indicating that subject velocity does influence the generation of GRF measurements in horses. Variation in subject velocity should be minimized when performing force-plate analysis in horses. PMID- 8720232 TI - Ultrastructure of selected struvite-containing urinary calculi from cats. AB - OBJECTIVE: To elucidate the ultrastructural details of struvite-containing urinary calculi from cats. DESIGN: Specimens studied were inclusive of the range of textures visible during preliminary analysis by use of a stereoscopic dissecting microscope. Textural types, which were used to infer crystal growth conditions, were differentiated with regard to crystal habit, crystal size, growth orientation, and primary porosity. SAMPLE POPULATION: Thirty specimens were selected from a collection of approximately 1,600 feline urinary calculi: 20 of these were composed entirely of struvite, and 10 consisted of struvite and calcium phosphate (apatite). PROCEDURE: Qualitative and quantitative analyses of specimens included use of plain and polarized light microscopy, x-ray diffractometry, scanning electron microscopy with backscattered electron imagery, x-ray fluorescence scans, and electron probe microanalysis. RESULTS: Four textural types were recognized among struvite calculi, whereas 2 textural types of struvite-apatite calculi were described. CONCLUSIONS: The presence of minute, well interconnected primary pores in struvite-containing urinary calculi from cats is an important feature, which may promote possible interaction of calculi with changes in urine composition. CLINICAL RELEVANCE: Primary porosity, which can facilitate interaction between the calculus and changing urine composition, may explain the efficacy of dietary or medicinal manipulations to promote the dissolution of struvite-containing uroliths from this species. PMID- 8720233 TI - Comparison of methods for sodium and potassium determination in llama urine. AB - OBJECTIVE: To compare results for sodium and potassium determination on llama urine, using flame emission spectrophotometry (flame photometry), atomic absorption spectrophotometry (AAS), indirect ion-selective electrode potentiometry (ISE), and direct ISE. DESIGN: Llama urine samples encompassing a wide range of electrolyte concentrations were analyzed for sodium and potassium concentrations, using 4 analytical methods, and results were compared statistically to assess correlation, bias, and potential interferents. SAMPLE POPULATION: 10 healthy male llamas. PROCEDURE: Urine specimens were obtained from llamas fitted with urine collection apparatus at defined intervals over a 24-hour period. Urine samples were centrifuged, and supernatants were frozen at -70 C until analysis. Analytical procedures were done, using standard laboratory protocols. Means, correlation coefficients, and bias were calculated, and differences were evaluated by ANOVA, with significance set at P < 0.05. RESULTS: There was strong correlation and good agreement among sodium values obtained by flame photometry, AAS, and indirect ISE. Sodium values obtained by use of direct ISE correlated poorly with other methods; urine is not an acceptable specimen for this method. Only AAS and indirect ISE had good correlation (r > 0.9) for potassium values. Data did not suggest presence of a potassium chelator in llama urine; urine potassium values measured by indirect ISE were significantly higher (by 150 to 200 mmol/L) than those measured by other methods. CLINICAL RELEVANCE: Urine electrolyte analysis in llamas resulted in less agreement between methods than is generally found for serum. Data collection for patient monitoring or research analysis should be restricted to a single method to avoid differences in results attributable to analytical variance. PMID- 8720234 TI - Effects of aging on the endodontic system, reserve crown, and roots of equine mandibular cheek teeth. AB - OBJECTIVE: To document age-related changes in the morphology of the endodontic system, reserve crown, and roots of equine mandibular cheek teeth. DESIGN: Equine mandibular cheek teeth from horses of various ages were compared, using radiography, x-ray computed tomography, and histologic examinations. SAMPLE POPULATION: 48 right hemi-mandibles from horses 2 to 9 years old. PROCEDURE: Hemi mandibles were radiographed, imaged by computed tomographic reconstruction, and reformatted. Histologic examination was used to identify and correlate tissue types. RESULTS: Permanent mandibular cheek teeth of the horse, at the time of eruption, consisted of an exposed crown and a reserve crown with a widely dilated apex. The endodontic system consisted of 5 or 6 pulp horns that connected to an expansive pulp in the reserve crown, which was confluent with the primordial pulp bulb surrounding the tooth's apex. At the time of eruption, mandibular cheek teeth did not have a distinct pulp chamber, roots, or evidence of root formation. However, within 2 years after eruption, mesial and distal roots and a pulp chamber were present. A distinct pulp chamber, communicating with the pulp horns and both root pulp canals, was identifiable for 4 to 5 years from the time of root formation. The endodontic system of cheek teeth, 6 to 8 years after eruption, consisted of 2 unattached compartments, made up of a root canal, pulp chamber, and 2 or 3 pulp horns. CLINICAL RELEVANCE: The age-related morphologic changes in equine mandibular cheek teeth have important implications for application of endodontic therapy in horses. PMID- 8720235 TI - Epidemiological characteristics of bovine clinical mastitis caused by Staphylococcus aureus and Escherichia coli studied by DNA fingerprinting. AB - OBJECTIVE: To study the epidemiology of clinical mastitis caused by Escherichia coli and Staphylococcus aureus by differentiating isolates with DNA fingerprinting techniques, using polymerase chain reaction. DESIGN: Milk samples were collected from cases of clinical mastitis in dairy cows. Escherichia coli and S aureus isolates from these cases were compared within and between cows and herds. SAMPLE POPULATION: Seven dairy herds with an average bulk milk somatic cell count < 150,000/ml, and incidence of cows with clinical mastitis of > 25%/y. PROCEDURE: Chromosomal DNA was isolated from E coli and S aureus strains isolated from cases of clinical mastitis, and amplified by polymerase chain reaction, using enterobacterial repetitive intergenic consensus primers for E coli and a random amplified polymorphic DNA primer for S aureus. Escherichia coli and S aureus strains were identified and differentiated, using their DNA polymorphism pattern. RESULTS: Multiple E coli genotypes were found in each of the herds. Persistent infections with E coli were sporadic. Only a limited number of different S aureus genotypes was found in each of the herds studied. Recurrent cases of S aureus mastitis were found in 25% of quarters with clinical S aureus mastitis. Comparing S aureus isolates from different herds indicated that 1 S aureus genotype was most prevalent. CONCLUSIONS: Because different quarters were infected with different genotypes, it was concluded that E coli is an environmental pathogen, and does not generally spread from quarter to quarter. The hypothesis that S aureus mastitis is a contagious disease, spreading from infected to uninfected quarters, could not be rejected. PMID- 8720236 TI - Transmission of two pseudorabies virus strains that differ in virulence and virus excretion in groups of vaccinated pigs. AB - OBJECTIVE: To determine whether 2 pseudorabies virus (PRV) strains that differ in virulence differ in transmission among vaccine strain 783-inoculated pigs. DESIGN: Three experiments were conducted, each with 2 groups of 10 pigs inoculated with the glycoprotein E-negative vaccine 783. In the first 2 experiments, half of one group was inoculated with the mildly virulent strain Sterksel, and half of the other group was inoculated with the virulent strain NIA 3. The remaining pigs were contact-exposed to the inoculated pigs. In the third experiment, 2 pigs in each group were inoculated with strain NIA-3, and the other 8 pigs in each group were contact-exposed to these 2 pigs. ANIMALS: Specific pathogen-free pigs that were sero-negative for antibodies to PRV. PROCEDURE: Estimation of the transmission of virus in each group was based on a stochastic model, in which the observable variable was the number of contact infections. The sole parameter of the model was the reproduction ratio theta. A glycoprotein E ELISA was used to determine whether infection occurred. Virus excretion in the oropharyngeal fluid was determined by plaque assay to measure infectivity. RESULTS: Vaccinated pigs had a significant difference in virus excretion between the mildly virulent and virulent strains. Sterksel strain-inoculated pigs shed significantly more virus than did NIA-3 strain-inoculated pigs. Despite this difference in virus excretion, the 2 strains did not differ in transmission. CONCLUSIONS: Both PRV strains would be eliminated from a population of vaccine strain 783-inoculated pigs. PMID- 8720237 TI - Infection and cross-protection studies of winter dysentery and calf diarrhea bovine coronavirus strains in colostrum-deprived and gnotobiotic calves. AB - OBJECTIVE: To investigate in vitro antigenic relations, in vivo cross-protection, and isotype antibody responses to a winter dysentery (WD) and calf diarrhea strain of bovine coronavirus (BCV). DESIGN AND ANIMALS: Gnotobiotic and colostrum deprived calves were inoculated oronasally with a WD (DBA) or a calf diarrhea (DB2) BCV, and were challenge exposed with the heterologous BCV. PROCEDURE: Nasal swab and feces specimens and blood samples were collected. Fecal and nasal specimens were assayed for BCV shedding by antigen-capture ELISA or immune electron microscopy. Bovine coronavirus antigens were detected in nasal epithelial cells by immunofluorescence. Antibody titers to BCV in serum were assayed by virus neutralization (VN), and BCV antibody isotype titers in feces and sera were quantitated by ELISA. RESULTS: All calves developed diarrhea and shed BCV nasally and in feces, then recovered and were protected from BCV associated diarrhea after challenge exposure with the heterologous BCV. After challenge exposure with either strain, fecal shedding of DBA was detected in 1 of 4 calves and nasal shedding of DB2 was detected in 2 of 4 calves. Immunoglobulin M was the principal coproantibody to BCV early, followed predominantly by IgA. Immunoglobulin G1 coproantibody titers to BCV were low, but increased after challenge exposure. Immunoglobulin G1 antibodies were predominant in serum. After challenge exposure, all serum antibody isotype titers increased except IgG2. The VN antibody responses paralleled serum IgG1 antibody responses. CONCLUSIONS AND CLINICAL RELEVANCE: Immunoglobulin A coproantibodies at challenge exposure were associated with protection against diarrhea. Nasal shedding of BCV after challenge exposure confirmed field data documenting reinfection of the respiratory tract of cattle, suggesting that, in closed herds, respiratory tract infections constitute a source of BCV transmission to cows (WD) or young calves. PMID- 8720238 TI - Prevalence of coagulase-positive staphylococci, other than Staphylococcus aureus, in bovine mastitis. AB - OBJECTIVE: To determine prevalence and relevance of coagulase-positive Staphylococcus hyicus and S intermedius intramammary infections (IMI) in dairy cows and determine the ability of the 4-hour tube coagulase (TC) test to differentiate the coagulase-positive staphylococci (CPS). DESIGN: Prevalence of CPS was determined for primiparous cows (point prevalence and prevalence at first parturition) and multiparous cows (point prevalence) of 2 herd groups: < 6% CPS IMI prevalence = low prevalence (LP); > 10% CPS IMI prevalence = high prevalence (HP). SAMPLE POPULATION: For prevalence, cows of 22 dairy herds. For TC, 1,038 CPS strains isolated from cow milk. PROCEDURE: Speciation of CPS from aseptically collected composite milk samples was performed. Coagulase-positive isolates from 4 cow groups were tested for their ability to coagulate rabbit plasma by 4 hours: LP and HP primiparous cows at parturition, and LP and HP cows any time after first parturition. RESULTS: Of 487 CPS in the prevalence study, 82.1% were S aureus, 17.7% were coagulase-positive S hyicus, and 0.2% were S intermedius. Of all CPS IMI in LP herds, 34% were coagulase-positive S hyicus; of all CPS IMI in HP herds, 9% were coagulase-positive S hyicus. Coagulase-positive S hyicus appeared to persist to the end of lactation in 4 cows (mean linear somatic cell count = 3.7). The TC test was > or = 97% sensitive, < or = 33% specific, and had a predictive value positive range of 60 to 97% for S aureus isolates. CONCLUSION: Coagulase-positive S hyicus appears capable of inducing chronic, low-grade IMI. Staphylococcus intermedius does not appear to be an important mastitis pathogen. The TC test is not valid to use as the sole method to differentiate CPS species. PMID- 8720239 TI - Preliminary evaluation of antimicrobial agents for treatment of Leptospira interrogans serovar pomona infection in hamsters and swine. AB - OBJECTIVE: To evaluate antimicrobial agents for treatment of models of acute and persistent leptospirosis caused by Leptospira interrogans serovar pomona. DESIGN: Randomized trials comparing dosages and regimens of various antimicrobial agents for treatment of acute and persistent leptospirosis. ANIMALS: 245 Golden hamsters to model acute leptospirosis and 121 mixed-breed swine to model persistent leptospirosis. PROCEDURE: Hamsters and swine were inoculated with L interrogans serovar pomona. Antimicrobial agents were given to hamsters for 3 or 5 days after inoculation, with necropsy at 14 days after inoculation. Swine were treated for 1, 3, or 5 days beginning at 3 weeks after inoculation, and were necropsied 7 to 10 days after completion of antimicrobial agent treatment. Hamster tissue and swine tissue and urine specimens were examined by culture, fluorescent antibody testing, and histologic examination for presence of leptospires. RESULTS: All untreated control hamsters became infected and manifested clinical signs and lesions of acute leptospirosis. Leptospires were not detected in hamsters treated with dihydrostreptomycin/penicillin G (25 mg/kg of body weight). Administration of ampicillin at all dosages reduced the number of hamsters infected, as confirmed at necropsy; the other agents tested required dosages greater than label recommendations to reduce the number infected. All untreated control swine became infected and shed leptospires in urine through the time of necropsy. Leptospires were not detected in kidneys or urine of swine treated with dihydrostreptomycin/penicillin G (25 mg/kg) for 1, 3, or 5 days, or in swine treated with oxytetracycline (40 mg/kg for 3 or 5 days), tylosin (44 mg/kg for 5 days), or erythromycin (25 mg/kg for 5 days). Treatment with ceftiofur and ampicillin was not effective in elimination of L interrogans serovar pomona in swine. CONCLUSIONS: Dihydrostreptomycin/penicillin G is effective for treatment of acute and persistent leptospirosis. Differences between the effectiveness of antimicrobial agents in the acute and persistent model of leptospirosis emphasize the importance of using the appropriate model for treatment evaluation. Antimicrobial agents evaluated for treatment of persistent leptospirosis in swine required the use of dosages above those recommended by the manufacturer. CLINICAL RELEVANCE: Use of antimicrobial agents at extra-label dosages for treatment of persistent leptospirosis may cause residue problems in food animals; however, these regimens may be useful for treatment of breeding stock or animals destined for import/export. PMID- 8720240 TI - Detection of strain-specific antigenic epitopes on the lipo-oligosaccharide of Haemophilus parasuis by use of monoclonal and polyclonal antibodies. AB - OBJECTIVE: To investigate the antigenic diversity of lipo-oligosaccharides of Haemophilus parasuis. PROCEDURES: Immunoblot assays were done with monoclonal and polyclonal antibodies on whole-cell lysates. Individual colonies of H parasuis strains H 54, H 53, and H 128 were tested for reactivity with lipo oligosaccharide-specific monoclonal antibodies after a single passage on chocolate agar, and colonies of strain H 54 were analyzed after 10 passages. Colony blot tests were used to screen H parasuis strains for spontaneously occurring antigenic variation in their lipo-oligosaccharides. RESULTS: Eight H parasuis strains were separated into 4 lipo-oligosaccharide serovars on the basis of immunoblot reactions with 3 polyclonal rabbit antisera. Nine monoclonal antibodies against lipo-oligosaccharides of a lipo-oligosaccharide-serovar I strain reacted with all tested serovar I strains but failed to react with other H parasuis strains. CONCLUSIONS: Variations in the antigenic reactivity after 1 or 10 passages on chocolate agar were not observed. The serovar I lipo oligosaccharide strains included virulent as well as avirulent H parasuis strains, indicating that these epitopes do not correlate directly with virulence properties of H parasuis. PMID- 8720241 TI - Demonstration of synergistic effects of sulfonamides and dihydrofolate reductase/thymidylate synthase inhibitors against Neospora caninum tachyzoites in cultured cells, and characterization of mutants resistant to pyrimethamine. AB - OBJECTIVE: To examine the efficacies of combinations of 7 sulfonamides and 5 dihydrofolate reductase/thymidylate synthase (DHFR/TS) inhibitors against tachyzoites of Neospora caninum in cultured cells. Mutant tachyzoites that were resistant to pyrimethamine were produced and examined for resistance to other DHFR/TS inhibitors. DESIGN AND PROCEDURES: After 5 days of treatment, a cell culture flask lesion-based assay was used to determine efficacies of combinations of sulfonamides and DHFR/TS inhibitors against N caninum tachyzoites and to evaluate the sensitivity of pyrimethamine-resistant mutants of N caninum to test agents. Cultured cells that were infected with the appropriate strains of N caninum and treated or not treated (controls) with test agents were examined. Mutations were induced by chemical mutagenesis with N-methyl-N'-nitro-N nitrosoguanidine or by selection for growth in permissive concentration of pyrimethamine. RESULTS: Synergism was detected for combinations of pyrimethamine, ormetoprim, trimethoprim, or diaveridine with the sulfonamides. Methotrexate did not have improved efficacy when combined with sulfonamides. Two mutants were produced that were resistant to pyrimethamine. Both mutants were resistant to other DHFR/TS inhibitors. Both mutants remained resistant to pyrimethamine in the absence of continuous exposure to the agent, indicating that the induced resistance was stable. Synergism was detected for combinations of DHFR/TS inhibitors and sulfonamides against these pyrimethamine-resistant mutants. CONCLUSIONS: Combinations of suboptimal concentrations of sulfonamides with suboptimal concentrations of DHFR/TS inhibitors results in improved efficacy of the agents in a cell culture assay. Stable resistance to pyrimethamine can be induced in N caninum tachyzoites by use of chemical mutagenesis or by selection. CLINICAL RELEVANCE: In vitro evidence indicated that combination treatment, using sulfonamides and DHFR/TS inhibitors, may be effective in treating neosporosis. PMID- 8720242 TI - Ampicillin and amoxicillin residue detection in milk, using microbial receptor assay (Charm II) and liquid chromatography methods, after extra-label administration of the drugs to lactating cows. AB - OBJECTIVE: A microbial receptor assay method (MRAM; Charm II test) for beta lactam antibiotics and a liquid chromatography (LC) method with a detection limit of 2 to 5 ppb were evaluated for detection of ampicillin or amoxicillin residues in milk samples from individual cows. DESIGN: The MRAM was compared to the LC in 2 respects. Measured concentrations of drugs were compared, as well as the classification of samples relative to the FDA tolerance value of 10 ppb. ANIMALS: A total of 6 clinically normal lactating Holstein cows were used per drug. PROCEDURE: Ampicillin trihydrate or amoxicillin trihydrate was administered at an extra-label dosage of 22 mg/kg of body weight, IM, once to each of 6 cows/drug. Milk samples were collected at milkings prior to and for 156 hours after drug administration. Drug concentrations in milk samples from individual cows were determined by use of the MRAM and LC tests. Additionally, the classification of milk samples relative to the presence or absence of residues above the FDA tolerance value was determined. Pharmacokinetic analysis was performed on derived milk drug concentrations. RESULTS: Concentration of ampicillin in milk samples from all cows was < 10 ppb by the MRAM and LC methods by the fourth milking (48 hours) after treatment with ampicillin. Values were < 10 ppb by both methods for all cows treated with amoxicillin by the sixth milking (72 hours) after treatment. For individual milk samples, significant differences were found between test methods in the proportion of positive (failing) tests; the MRAM had a higher proportion of presumptive positives. CONCLUSIONS: Even at an extra-label dosage of 22 mg/kg, IM, milk residues > 10 ppb (the FDA tolerance value) were not detected beyond the label milk withholding times for ampicillin (48 hours) and amoxicillin (96 hours). When used for testing milk of individual cows by the control point procedure, the MRAM had a tendency to give presumptive positive test results for milk samples containing < 10 ppb ampicillin or amoxicillin as determined by LC. PMID- 8720243 TI - Effects of topical administration of 0.5% apraclonidine on intraocular pressure, pupil size, and heart rate in clinically normal dogs. AB - OBJECTIVE: To evaluate the effect of 0.5% apraclonidine on intraocular pressure (IOP), pupil size, and heart rate in clinically normal dogs. DESIGN: Randomized masked saline-controlled case study. ANIMALS: Nine clinically normal conditioned adult dogs of either sex. PROCEDURE: Normal diurnal variation in IOP, pupil size, and resting heart rate were determined from 7 AM to 7 PM (day 1). These measurements were repeated on day 2 after topical application of 60 microliters of 0.5% apraclonidine to 1 randomly chosen eye of each dog. The contralateral eye received saline solution. RESULTS: Compared with the saline-treated fellow eye, mean IOP in the apraclonidine-treated eye was significantly reduced (3.0 mm of Hg, 16%) 8 hours after treatment. Because of mild day-to-day variations in IOP, however, IOP in the apraclonidine-treated eye on day 2 was not significantly different from day-1 baseline values obtained from the same eye. Significant mydriasis (2.1 mm, 29.7%), persisting for up to 8 hours, occurred in apraclonidine-treated eyes. Although apraclonidine did not significantly alter heart rate when all 9 dogs were viewed as a group, 4 dogs experienced a 9 to 19.5% reduction in heart rate 2 hours after treatment. Mild blanching of the conjunctiva occurred in apraclonidine-treated eyes. CONCLUSIONS: Apraclonidine lowered IOP and, in contrast to cats where it causes miosis, induced mydriasis in dogs. Although heart rate generally is unchanged, it may be reduced in select individuals. CLINICAL RELEVANCE: Topically applied 0.5% apraclonidine may be a useful adjunct to other antiglaucoma treatment modalities in dogs, but is unlikely to be effective as the sole agent in most forms of canine glaucoma. PMID- 8720244 TI - Effects of topical administration of 0.5% apraclonidine on intraocular pressure, pupil size, and heart rate in clinically normal cats. AB - OBJECTIVE: To evaluate the effect of a topical alpha 2-agonist, 0.5% apraclonidine, on intraocular pressure (IOP), pupil size, and heart rate in clinically normal cats. DESIGN: Randomized masked saline-controlled case study. ANIMALS: Nine clinically normal conditioned adult cats of either sex. PROCEDURE: Normal diurnal variation in IOP, pupil size, and resting heart rate was determined from 7 AM to 7 PM (day 1). On day 2, the same measurements were made after the topical application of 30 microliters of 0.5% apraclonidine to 1 randomly chosen eye of each cat. The contralateral eye received saline solution. RESULTS: Apraclonidine lowered IOP a mean of 4.8 mm of Hg (24.0%) when compared with IOP in the fellow saline-treated eye 6 hours after treatment. Unilateral administration of the drug also may have reduced IOP in both eyes. Pupil size was reduced a mean 46% in only the apraclonidine-treated eye, and miosis persisted for up to 24 hours. Heart rate was significantly lower (11.8%) at 3 hours, and 8 of 9 cats vomited after topical administration of the drug. Mild blanching of the conjunctiva occurred in all apraclonidine-treated eyes. CONCLUSIONS: Apraclonidine reduced IOP, pupil size, and resting heart rate in clinically normal cats, but also induced undesirable systemic side-effects. In contrast to dogs where the drug induced mydriasis, apraclonidine caused miosis in cats. CLINICAL RELEVANCE: The current commercially available formulation of topical 0.5% apraclonidine is too toxic for clinical use in cats. Reformulation, or identification of a less toxic congener, may result in a clinically useful antiglaucoma preparation for cats. PMID- 8720245 TI - Comparative cardiopulmonary effects of intramuscularly administered etorphine and carfentanil in goats. AB - OBJECTIVE: To determine comparative cardiopulmonary effects of IM administered etorphine and carfentanil in goats. ANIMALS: Seven clinically normal adult female goats. DESIGN: Each goat received at least 9 drug treatments (etorphine HCl, 5 [twice], 10, 20, and 40 and carfentanil citrate, 5, 10, 20 and 40 micrograms/kg of body weight), with a minimal 2-day interval between trials. Although drug dosages were randomized, etorphine and carfentanil treatments were alternated. To assess for drug tolerance, the first and last treatments always were etorphine (5 micrograms/kg). PROCEDURE: All goats were instrumented for long-term cardiopulmonary variable data collection. RESULTS: Both drugs induced rapid catatonic immobilization, characterized by limb and neck hyperextension, with occasional vocalization and bruxation. Etorphine elicited transient violent struggling and vocalization immediately. Time to immobilization appeared dose dependent, and was more rapid with carfentanil (< or = 5 minutes) than etorphine (5 to 10 minutes) at all dosages. Recovery to standing occurred earlier for etorphine (1 to 2 hours) than carfentanil (> 2 hours) at all dosages. Both drugs at all dosages significantly (P < or = 0.05) increased systemic and left ventricular (LV) end-diastolic pressures, LV peak negative dP/dt, total peripheral resistance (TPR), hemoglobin concentration, and left atrial (LA) and pulmonary O2 contents. They also significantly decreased heart and respiration rates, and TPR. A significant increase was observed at some dosages for LV stroke volume and index, LV peak positive dP/dt, mean pulmonary artery pressure, PaO2, pulmonary artery oxygen partial pressure, PaCO2, pulmonary mixed venous carbon dioxide partial pressure, LA hemoglobin saturation, LA transport index, and body temperature. Pulmonary and systemic mixed venous carbon dioxide and oxygen contents were significantly decreased at some dosages. CONCLUSIONS: Intramuscularly administered etorphine and carfentanil induce hypertension, bradycardia, and bradypnea in goats. The hypertension appears attributable to an increase in TPR. CLINICAL RELEVANCE: Although the cardiopulmonary effects of carfentanil occurred more rapidly, these effects were similar in magnitude for etorphine and carfentanil over the evaluated dosage range. PMID- 8720246 TI - Use of hypertonic saline-dextran solution to resuscitate hypovolemic calves with diarrhea. AB - OBJECTIVE: To determine effectiveness of a new and practical method for fluid resuscitation of dehydrated diarrheic calves. DESIGN: Animals randomly allocated to 4 groups with appropriate controls. ANIMALS: 16 healthy male dairy calves, 3 to 6 days old. PROCEDURE: After instrumentation and recording baseline data, diarrhea and hypovolemia were induced by administering milk replacer (33 ml/kg of body weight) and isotonic sucrose solution (2 g of sucrose in 19.5 ml of water/kg, PO) every 8 hours, and furosemide (2 mg/kg, IM) every 4 to 8 hours. Administration of milk replacer and furosemide was discontinued when calves became 6% dehydrated. Calves were then randomly allocated as: control (no treatment); hypertonic saline-dextran (HSD) solution (4 ml/kg, 2,400 mOsm/L NaCl in 6% dextran-70, administered once over 4 minutes, IV); isotonic alkalinizing oral electrolyte solution (55 ml/kg, PO, q 8 h); and HSD-oral electrolyte solution (combination of HSD and oral treatments). Calves were monitored for 24 hours after treatment. RESULTS: Significant changes included moderate dehydration (8% body weight), marked lethargy, decreased cardiac output and plasma volume, and increased blood lactate concentration, hematocrit, and serum concentrations of albumin, creatinine, sodium, and phosphate. Control calves continued to be lethargic and dehydrated, with significant increases in hematocrit and serum creatinine concentration. Increase in cardiac output and plasma volume was transient in the HSD group and waned by 2 to 8 hours after treatment. Oral electrolyte fluid administration caused slow and sustained increase in cardiac output and plasma volume, and decrease in heart rate, blood lactate concentration, and hematocrit. Combined administration of HSD-oral electrolyte solution caused immediate and sustained increase in cardiac output and plasma volume, and decrease in heart rate, blood lactate concentration, and hematocrit. CONCLUSIONS AND CLINICAL RELEVANCE: Treatment of hypovolemic diarrheic calves with IV HSD and oral electrolyte solution is superior to administration of either solution alone. PMID- 8720247 TI - Evaluation of a single injection method, using iohexol, for estimating glomerular filtration rate in cats and dogs. AB - OBJECTIVE: To evaluate the utility of a method for estimating glomerular filtration rate (GFR) after single IV administration of iohexol. DESIGN: The plasma clearance of iodine (PCI), taken as the quotient of the administered dose of iodine (300 to 600 mg of l/kg of body weight) divided by the area under the plasma iodine concentration versus time curve determined by 4 methods (PCI1 PCI4). The results for PCI were compared with simultaneously obtained values for the urinary clearance of exogenously administered creatinine (CCr), a widely accepted method for the measurement of GFR in cats and dogs. ANIMALS: Cats and dogs that were renal intact (n = 5 cats; n = 1 dog) or had renal mass reduced by partial nephrectomy (n = 5 cats; n = 7 dogs). RESULTS: Values for PCI were closely related (R2 values ranged from 0.947 to 0.992; P < 0.0001 in all cases) to CCr. Despite this close correlation between CCr and PCI, the 95% confidence interval for the difference between PCI3 and CCr included values that exceeded 1.4 ml/min/kg, which represents 50% of the mean value for CCr in renal-intact cats. CONCLUSIONS: Determination of PCI provided a reliable estimate of GFR in cats and dogs of this study. However, differences between 1 of the methods (PCI3) and CCr are clinically important, emphasizing the need to use more than simple linear regression analysis and correlation coefficients when attempting to validate new measurement techniques. CLINICAL RELEVANCE: The determination of PCI provided a reliable estimate of GFR in cats and dogs of this study. PMID- 8720248 TI - Effects of tendon grip technique (frozen versus unfrozen) on in vitro surface strain measurements of the equine deep digital flexor tendon. AB - OBJECTIVE: To determine effects of tendon grip technique on in vitro surface strain measurements of equine deep digital flexor tendon (DDFT) when loaded in tension. SAMPLE POPULATION: 12 hind limb DDFT from 8 adult horses (mean age, 9.8 years [range, 4.5 to 17 years]; mean body weight, 472 kg [range, 450 to 509 kg]), with no clinical evidence of hind limb lameness. DESIGN AND PROCEDURE: After calibration, liquid mercury strain gauges were sutured to plantar surfaces of the tendons at distal (position 1), middle (position 2), and proximal (position 3) metatarsal regions. Each tendon was affixed to a materials testing machine (distally by the distal phalanx, and proximally by a metal clamp), and loaded once in tension, at a strain rate of 2.8 to 3.0%/s, to 3% clamp displacement. Liquid nitrogen was used to freeze the proximal ends of 6 tendons before placement in the clamp. Proximal ends of the remaining 6 tendons were left unfrozen. Surface strains were determined with the aid of low-resistance bridge circuits. Data were analyzed at time points corresponding to 1, 2, and 3% clamp displacement. RESULTS: Mean surface strains of tendons with frozen ends were significantly greater than those for tendons with unfrozen ends, and closer to strain measured by clamp displacement, at 1 and 2% clamp displacement. This difference was present but not significant at 3% clamp displacement. Mean local surface strain was not significantly different between the 3 gauge positions. CONCLUSIONS: Freezing tendon ends is a useful technique to increase measured surface strains in equine DDFT, thereby making them closer to universal strain as measured by clamp displacement, and, thus, more likely to represent true surface strain. PMID- 8720249 TI - Effect of craniectomy/durotomy alone and in combination with hyperventilation, diuretics, and corticosteroids on intracranial pressure in clinically normal dogs. AB - OBJECTIVE: To determine the effect of craniectomy and durotomy on intracranial pressure (ICP) in clinically normal dogs. DESIGN: Two-part study (experiments A and B) involving craniectomy and durotomy, with and without treatments to lower ICP. ANIMALS: Six (experiment A) and 7 (experiment B) healthy dogs. PROCEDURE: In experiment A, craniectomy was performed in combination with durotomy, diuretic administration, methylprednisolone sodium succinate administration, and hyperventilation, and effect of these manipulations on ICP was determined. In experiment B, dogs had only craniectomy and durotomy without associated ICP lowering treatments. During both experiments, ICP was monitored throughout the surgical procedure with a fiber optic ICP monitoring device. RESULTS: Intracranial pressure decreased after the combination of craniectomy, durotomy, and other ICP-lowering treatments in dogs of experiment A. Similar magnitude of decrease in ICP was observed in dogs of experiment B after craniectomy and durotomy. CONCLUSIONS: Comparison of these experiments indicate that surgical removal of overlying skull and incision of the dura mater can significantly decrease ICP in clinically normal dogs. CLINICAL RELEVANCE: Craniectomy and durotomy may be useful as an adjunct treatment for increased ICP. PMID- 8720250 TI - Kinematic evaluation of gait in dogs with cranial cruciate ligament rupture. AB - OBJECTIVE: Noninvasive, computer-assisted, three-dimensional kinematic gait analysis was used to describe lameness in a chronic model of cranial cruciate ligament rupture (CCLR) in dogs. DESIGN: Hind limb lameness was evaluated prior to and at 1, 3, and 6 months after transection of the cranial cruciate ligament. ANIMALS: Seven clinically normal large dogs. PROCEDURE: Dynamic flexion and extension angles and angular velocities were calculated for the coxofemoral, femorotibial, and tarsal joints. Distance and temporal variables were determined. Essential Fourier coefficients were used to develop mean flexion extension curves for all joints and to compare changes in movement that developed with CCLR over time. RESULTS: Each joint had a characteristics pattern of flexion and extension movement that changed with CCLR. The femorotibial joint angle was more flexed throughout stance and early swing phase of stride and failed to extend in late stance. Angular velocity of the femorotibial joint was damped throughout stance phase, with extension velocity almost negligible. The coxofemoral and tarsal joint angles, in contrast to the femorotibial joint angle, were extended more during stance phase. These changes were documented as differences noted in the essential Fourier coefficients. Stride length and frequency also varied significantly after CCLR. CONCLUSIONS: Cranial cruciate ligament rupture affects movement of the coxofemoral and tarsal joints, as well as the femorotibial joint, in gait. A pattern of joint movement may be discerned in which the coxofemoral and tarsal joints compensate for the dysfunction of the femorotibial joint. CLINICAL RELEVANCE: Methods were developed that will improve objective evaluation of CCLR and its treatment in dogs. PMID- 8720251 TI - Induction of the low-molecular-weight stress protein HSP27 in organ-cultured normal human skin. AB - To examine the inducibility of 27-kD-molecular-weight heat shock protein (HSP27) in human skin, an indirect immunofluorescence (IF) study was performed on organ cultured normal human skin by using a monoclonal antibody specific for HSP27. After heat treatment at 45 degrees C for 1 h, nuclear IF was observed in the epidermal cells. When the organ-cultured skin explants were exposed to 10.0 micrograms/ml or 100.0 micrograms/ml 8-methoxypsoralen (8-MOP) for 1 h and then irradiated with UVA (320-400 nm), positive nuclear IF was also observed 6 h after UVA irradiation. Considering these results with the previous reports about HSP72, it appears that, in human skin, HSP27 as well as HSP72 plays an important role in resisting various environmental stresses. PMID- 8720252 TI - Two cases of metastatic malignant melanoma of the lower limb treated with hyperthermic isolated limb perfusion and concomitant infusion of either carboplatin or beta-interferon. AB - Two cases of metastatic malignant melanoma of the lower limb who were treated successfully with hyperthermic isolated limb perfusion are reported. One patient was infused with cisdiammine (1.1-cyclobutanedicarboxylate) platinum (II) (carboplatin, Paraplatin, Bristol-Myers Squibb Company, New Jersey, USA), and the other was infused with human natural beta-interferon (Feron, Toray, Tokyo, Japan), via the external iliac artery. The first case showed a remarkable suppression of the growth of multiple metastatic melanoma nodules associated with numerous melanophage infiltrations, as shown histopathologically after the operation. The patient's serum level of 5-S-cysteinyl dopa decreased for the two months following the treatment. In the second case, new formation of metastatic melanoma nodules was completely suppressed for up to 12 months following the operation. Analysis of immunological parameters showed that the number of peripheral CD8+ lymphocytes gradually and constantly increased after the operation, while that of CD4+ lymphocytes transiently increased and then returned to the pre-operative level. Natural killer activity transiently decreased to a slight degree 4 days after the operation and then returned to the pre-operative level 21 days after the operation. Side effects, such as nausea, vomiting and leg discomfort, were seen in the patient (Case 1) treated with carboplatin, but were completely reversible. These results suggest that hyperthermic isolated limb perfusion with concomitant infusion of carboplatin or beta-interferon is effective in suppressing the growth of metastatic malignant melanomas of the lower limb. PMID- 8720253 TI - Serum concentration of 5-S-cysteinyldopa in pediatric patients with giant pigmented nevi. AB - The serum concentrations of 5-S-cysteinyldopa (5-S-CD) in pediatric patients with giant pigmented nevi (GPN) were investigated and compared with those of pediatric patients with small- or medium-sized congenital nevi (CN), or non-melanocytic benign skin tumor (control group). Serum 5-S-CD levels in the GPN group (N = 21), particularly in patients less than 5 years old, were significantly higher (highest concentration 38.4, mean +/- S.D. 16.6 +/- 9.4 nmol/L) than those in the CN (N = 22) or control groups (N = 26). Serum 5-S-CD levels in the CN group were not significantly different from those in the control group. There was a significant correlation between the serum 5-S-CD level and size (surface area) of the GNP. There was a significant inverse correlation between the serum 5-S-CD level and age in the CN and control groups, but not in the GPN group. Serum 5-S CD levels were transiently elevated immediately after treatment of patients with GPN with a combination of skin abrasion and cryotherapy with solid carbon dioxide. These results suggest that serum 5-S-CD levels in the GPN group reflected the 5-S-CD derived from GPN, particularly in patients less than 5 years old. This indicates that melanogenesis may be accelerated in infant patients in the GPN group. PMID- 8720254 TI - Immunohistochemical study of cellular events in lesional skin during common virus infections. AB - Determination was made of epidermal Langerhans cell (LC) distribution and infiltrating cellular events in lesional skin during varicella zoster virus (VZV) infection, and the results were compared with those for herpes simplex (HS), measles, and rubella by immunohistochemical staining with cell surface markers. CD1a positive epidermal LCs increased in number, particularly in measles and rubella. The number of LCs was within the normal range or slightly increased in the epidermis of VZV infection. In herpes zoster (HZ) and varicella, HLA-DR positive epidermal cells were present in the basal part of the epidermis. In measles, HLA-DR positive cells aggregated in papular lesions. In measles and rubella, the number of HLA-DQ positive epidermal cells appeared to increase. In HS cases, CD11b (OKM1) positivity of the upper epidermal keratinocytes was quite pronounced, but not in the basal layer. CD8 positive suppressor/cytotoxic cells extensively infiltrated the dermis of HZ and varicella. Dermal infiltrates were identified as CD8 positive cell dominant in measles, HZ, and varicella. These results provide a partial explanation as to why cellular events in skin lesions act immunosuppressively. PMID- 8720255 TI - Anti-ribosomal P protein antibodies in a Japanese patient with systemic sclerosis. AB - A 56-year-old Japanese woman developed Raynaud's phenomenon and digital ulcerations at the age of 50. She showed rapid and diffuse skin sclerosis; visceral involvement was mild. She showed no symptoms or laboratory findings suggestive of other collagen diseases, including systemic lupus erythematosus and Sjogren's syndrome. Based on the clinical and histological findings, she was diagnosed as having diffuse cutaneous systemic sclerosis (SSc). The patient's serum produced nucleolar and cytoplasmic staining by indirect immunofluorescence analyses on HEp-2 cell substrate and reacted with P0, P1 and P2 proteins in immunoblotting using purified ribosomal antigens. She was negative for anti topoisomerase I, centromere, and U1RNP antibodies. Antiribosomal P protein antibodies are considered highly specific for systemic lupus erythematosus; this is the first case report of an SSc patient with anti-ribosomal P protein antibodies. The clinical features of SSc patients with these antibodies need to be clarified by an accumulation of cases. PMID- 8720256 TI - Acne fulminans coexisting with pyoderma gangrenosum-like eruptions and posterior scleritis. AB - A 30-year-old Japanese man presented with an erythematous, papulonodular eruption on his face and upper trunk, which rapidly progressed into severely exuberant, ulcerative lesions with a confluent tendency. His other symptoms included a high fever and general malaise. Complete blood cell counts showed neutrophilia with increased percentages of myelocytes and metayelocytes. The systemic administration of corticosteroids improved his skin lesions, leaving scars and milia. During tapering of corticosteroids, however, the patient developed posterior scleritis of his eyes and pyoderma gangrenosum-like eruptions on the lower legs. Our patient's history suggests that pyoderma gangrenosum-like eruptions and posterior scleritis are disorders associated with acne fulminans. PMID- 8720257 TI - Satisfactory remission achieved by PUVA therapy in Langerhans cell hisiocytosis in an elderly patient. AB - Langerhans cell histiocytosis is currently regarded as a reactive proliferative process of Langerhans cells rather than a malignancy. The disease is characterized by Langerhans cell infiltration of skin, lung, bone and other organs. We report a 74-year-old man with Langerhans cell histiocytosis who had generalized hemorrhagic and crusted papules. He also had diabetes insipidus. Because he did not have any severe constitutional symptoms or failure of vital organs, we applied topical PUVA treatment to his skin lesions, which responded well to the therapy. Diabetes insipidus, however, remained, in spite of X ray radiotherapy for the pituitary lesion. PMID- 8720258 TI - A low grade scirrhous carcinoma of eccrine gland origin in adolescence. AB - We report a rare occurrence of adnexal carcinoma in an adolescent. A 17-year-old Japanese girl had a gradually enlarging tumor on the elbow. Histopathologically, the exophytic tumor showed asymmetrical lobules of tumor embedded in a thickened collagenous stroma. The lobules were composed of various patterns of tumor cell nests ranging from tubular to solid forms. Invasive growth patterns were also observed in the stroma. Two types of the cells, cuboidal and elliptically polygonal, were noted. Immunohistochemistry revealed the presence of S-100 protein and cytokeratin 8 and the absence of gross cystic disease fluid protein 15 in the tumor cells. Electron microscopic observation showed a ductal or glandular differentiation and fragmented basal laminae around the nests. Light and electron microscopic findings of this tumor are suggestive of a low-grade sweat gland carcinoma. PMID- 8720259 TI - Solitary milialike idiopathic calcinosis cutis: a case unassociated with Down syndrome. AB - We report a unique case of solitary milialike idiopathic calcinosis cutis (MICC) in a healthy Korean woman, which is not associated with Down syndrome. This case of MICC would be a form of idiopathic calcinosis cutis, which can be solitary or multiple, sporadic or associated with Down syndrome. PMID- 8720260 TI - Solitary piloleiomyoma with an unusual clinical appearance. AB - We present a 66-year-old man with a solitary piloleiomyoma on the forehead. The lesion had unusual clinical features; it was a crusted and dome-shaped nodule, which exhibited rapid growth. Histology revealed the typical features of piloleiomyoma. The unusual clinical features may have been due to its unique location, in which the downward growth of the tumor was prevented by the underlying frontal bone. PMID- 8720261 TI - Solitary neurilemmoma of the face. AB - A fourteen-year-old male with solitary neurilemmoma over his left cheek is described. Clinically, it was mistaken for molluscum contagiosum. The case is reported for its rarity and unique features. PMID- 8720262 TI - Unusual cases of syringoma of the forehead. AB - Syringoma involving the forehead is rare and can be confused with other diseases that are common on the forehead. Therefore, we describe here three patients with syringoma who developed numerous skin colored papules on the forehead. To our knowledge, syringoma of the forehead was reported previously only once by Tagami et al. PMID- 8720263 TI - Association of generalized pustular psoriasis, Sjogren syndrome, and Hashimoto's thyroiditis. PMID- 8720264 TI - [Future research for the treatment of intractable diseases]. PMID- 8720265 TI - [Comparison between the arthroscopic and open synovectomies for rheumatoid knee- a retrospective and random study on the results of the two methods]. AB - Knee joints affected by rheumatoid arthritis (RA) were randomized to either arthroscopic or open synovectomy, and the postoperative results were compared retrospectively. The materials consisted of 58 patients with 75 surgically treated knees. Arthroscopic synovectomy was performed on 37 knees of 30 patients, who were followed for five years and four months on average. Open synovectomy was performed on 38 knees of 28 patients, who were followed for five years and five months on average. The results were evaluated for ; recurrence of synovitis ; duration of the symptom-free period ; progressive bone changes on radiography ; range of motion (ROM) ; duration of hospital stay ; duration of surgery. Due to the less invasive surgical procedure, arthroscopic synovectomy resulted in less post operative pain, and there was no restriction of ROM after surgery, no need for postoperative rehabilitation, shorter hospital stay, lower incidence of osteoarthritic changes, and earlier return to the daily life. However, as synovial membrane removal was unavoidably insufficient, the effect of surgery lasted for only 18 months on average and recurrence of synovitis was observed in 73 % of patients. Open synovectomy meant relatively greater invasion and postoperative pain. Due to the restricted ROM in some cases, postoperative treatment was required and the hospital stay tended to be prolonged. Nevertheless, the recurrence rate was only 37 % during follow-up and the effect of surgery persisted for an average of 75 months, much better than that of arthroscopic synovectomy. In the treatment of RA knees, the indications for arthroscopic and open synovectomies should be carefully studied in the light of the merits and for demerits of each procedure. PMID- 8720266 TI - [What are IgG antibodies to single-stranded DNA measured by enzyme immunoassay- re-evaluation of their clinical significance]. AB - Enzyme immunoassay (EIA) to detect IgG antibodies to single-stranded DNA (ssDNA EIA) and EIA to detect anti double-stranded DNA antibodies (dsDNA EIA) have been developed and widely used in Japan. However, antigenecity of ssDNA and independence of anti ssDNA antibodies, and their clinical significance are still obscure. IgG anti dsDNA and ssDNA antibodies were simultaneously measured by dsDNA EIA and ssDNA EIA in sera of two hundred and eighty five patients from systemic lupus erythematosus (SLE) including other connective tissue diseases. Both EIAs were specific for SLE, although low levels of anti ssDNA antibodies were also detected in several other connective tissue diseases. Significant correlation was observed between two EIAs. All sera were divided into three group : 1 ) both EIAs were negative, 2 ) only ssDNA EIA was positive and 3 ) both EIAs were positive and in most sera ssDNA EIA levels were higher than dsDNA EIA levels. No sera were found which reacted only with dsDNA EIA or which reacted more strongly with dsDNA EIA than with ssDNA EIA. Both EIA titers were inhibited by dsDNA or ssDNA antigens by competitive inhipition study, but inhibition rate by ssDNA was higher than that by dsDNA. Previous reports by others that ssDNA EIA could be utilized to detect early stage of active SLE were not supported in this study. All these results suggested that most of what is called anti ssDNA antibedies detected by ssDNA EIA are not antibodies to purine and pyrimidine bases, but they might be anti dsDNA antibodies with low avidity which will react phosphate-ribose backbones or internal duplex structures in the ssDNA antigen. Only use of dsDNA-EIA is, thus, recommended for clinical purpose. PMID- 8720267 TI - [An autopsy case of microscopic polyarteritis nodosa resembling Schoenlein-Henoch purpura]. AB - A 86 year-old woman was re-admitted because of purpura of her upper and lower extremities, abdominal pain and blood stools. Seven weeks previously, she underwent a gastrectomy for gastric cancer. After re-admission, proteinuria and hematuria were noted, and the serum creatinine level increased. Two months, after the onset of purpura, she died of pneumonia. On autopsy examination, fibrinoid vasculitis of acute inflammatory stage (II) at small arteries and/or arterioles in the bladder, rectum, lungs, spleen and crescentic glomerulonephritis without immune deposits were observed. A diagnosis of microscopic polyarteritis nodosa (M PN) was made based on these clinical and histological findings. M-PN refers to systemic vaculitis with segmental necrotizing glomerulonephritis. However, this condition may be difficult to diagnose because vasculitis such as Scholein-Henoch purpura (SHP) and/or hypersensitivity angitis, diseases in which the small arteries and arteroles are mainly affected, occasionally bears a clinical and histological resemblance to M-PN. Because differential diagnosis from SHP was required, this case provided abundant suggestions with regard to the entity of M PN. PMID- 8720268 TI - [A case of Sjogren's syndrome and systemic lupus erythematosus complicated with necrotizing angiitis of the gallbladder]. AB - A 71 year-old man with Sjogren's syndrome and systemic lupus erythematosus was admitted to our hospital because of abdominal pain and fever. Laboratory investigation showed an ESR of 76 mm/hr, elevation of CRP and marked leukocytosis. Abdominal ultrasound and CT scan showed enlargement of the gallbladder and intravesicular sludge without stones. With a clinical diagnosis of acute cholecystitis and pan-peritonitis, the surgical procedures including cholecystectomy were performed. Histological examination of the gallbladder showed a presence of necrotizing angiitis of small arteries. The postoperative course was uneventful with daily administration of 10 mg of prednisolone. PMID- 8720269 TI - [A case of Behcet's disease associated with aortic regurgitation and nephrotic syndrome]. AB - A 50-year-old housewife with a main complaint of dyspnea was admitted to our hospital in September, 1992. In 1975, the patient developed aphthous stomatitis, genital aphthous ulcers, uveitis and erythema nodosum appeared and diagnosed as having Behcet's disease at Department of Dermatology of our hospital. Aortic regurgitation was pointed out in 1984. Hyperlipidemia appeared in January and Oral prednisolone was started in May, 1985. From April in 1987, proteinuria appeared. From January, 1988, urinal protein was greater than 300 mg/dl. Orthopnea appeared in July, 1992. Physical examinations revealed systolic and diastolic murmur and pretibial edema. Laboratory findings showed proteinuria (3.8 g/day) and hypoproteinemia (serum total protein 5.1 g/dl, serum albumin 3.1 g/dl). Microscopic findings of renal biopsy revealed mesangial proliferative glomerulonephritis and arteriosclerosis. Immunofluorescent studies demonstrated deposits of Apo B in the basement membrane and the mesangium. She was discharged from our hospital as proteinuria was decreased after a combination treatment with camostat mesilate 600 mg and SAIREITO 9.0 g. This patient developed nephrotic syndrome, which was caused by hypertension and hyperlipidemia in addition to Behcet's disease. This case is noteworthy because Behcet's disease is rarely complicated with nephrotic syndrome. PMID- 8720270 TI - [A case of polymyositis presenting histological picture of bronchiolitis obliterans organizing pneumonia with transbronchial lung biopsy specimens]. AB - A 69-year-old woman was admitted to our hospital because of slight fever, general fatigue, joint pain and proximal muscle weakness. Severe elevation of serum enzyme levels of CPK, transaminase and aldolase was noted. The chest roentgengram showed diffuse reticular and nodular infiltrates. Histological examination of the transbronchial lung biopsy specimens revealed alveolitis and organizing pneumonia. Daily administration of 80 mg predonisolone was effective for both lung findings and myositis. PMID- 8720271 TI - [A case of rheumatoid arthritis associated with pernicious anemia and bronchiolitis obliterans organizing pneumonia]. AB - A case of rheumateid arthritis (RA) with pernicious anemia (PA) and wandering multiple patchy densities in bilateral lung fields is reported. A 72-year-old woman was hospitalized in February 1994, because of cough. She had already advanced RA (Class IV, Stage IV). She showed macrocytic and hyperchromic anemia as follows ; red-cell count (RBC), 176 x 10(4)/microliters; hemoglobin (Hb),7.2 g/dl; hematocrit (Ht), 21.0% ; MCV, 119.3 fl; and MCH, 40.9 pg. Chest roentgenogram revealed multiple patchy densities in bilateral lung fields and there was no response to the administration of antibiotic agents. From these clinical pictures bronchiolitis obliterans organizing pneumonia (BOOP) was highly suspected. After steroid injection into the joint space, the abnormal lung shadows disappeared. Anemia had been recovering spontaneously, but recurred in July. The results of blood examination were as follows ; RBC, 162 x 10(4)/microliters; Hb, 6.7ng/dl; Ht, 19.1%; MCV, 117.9 fl; and MCH, 41.4 pg. Anti intrinsic factor antibody was positive. The level of serum vitamin B12 was low, 76 pg/ml. Sternal bone marrow aspiration showed magaloblastic changes with hypersegmentation of granulocytes. PA was diagnosed and improvement was noted after the intramuscular administration of vitamin B12. Subjective symptoms based on RA did not change during the clinical course. It is suggested that the pathogenesis about the combination of RA, BOOP and PA is related to common immunological abnormalities in our patient. A case of RA with PA and BOOP has not been reported previously, thus this case is considered clinically valuable. PMID- 8720272 TI - [Systemic sclerosis with various gastrointestinal problems including pneumoperitoneum, pneumatosis cystoides intestinalis and malabsorption syndrome]. AB - We describe here an experience of successful treatment of systemic sclerosis (SSc) complicated with various gastrointestinal (GI) problems including pneumoperitoneum, pneumatosis cystoides intestinalis and malabsorption syndrome. A 35-year-old female had developed selerodactyly since February, 1990. She had been treated under the diagnosis of SSc at other hospital. She had required several hospitalizations because of nausea, vomitting and abdominal distension, but her GI symptoms had gradually deteriorated. In April 1993, she was referred to our hospital and admitted for the treatment of her GI problems. On admission, she had systemic cutaneous sclerosis and marked abdominal distension without peritoneal signs was recognized. Chest and abdominal roentgenograms demonstrated massive free air under the diaphragm, marked dilation of small and large bowels, and multiple intestinal cysts (pneumatosis cystoides intestinalis ; PCI). We treated her GI problems with various modalities combined with medications, oxygen breathing, intravenous hyperalimentation and hyperbaric oxygen therapy. Pneumoperitoneum and PCI had disappeared after 8 courses of hyperbaric oxygen therapy and her GI symptoms had been well controled by intravenous hyper alimentation. Thereafter, she has been on intermittent parenteral nutrition through subcutaneous port inplantation. During the courses of this treatment, she developed an episode of Wernicke-Kolsakoff (W-K) syndrome which was considered to associate with malabsorption syndrome. The W-K syndrome had recovered by intravenous administration of vitamin B1. PMID- 8720273 TI - [Antineutrophil cytoplasmic antibody (ANCA) and vasculitis]. PMID- 8720274 TI - [Disruption of the cervical spine due to rheumatoid arthritis and its treatment]. PMID- 8720275 TI - Occupational and environmental medicine in the United Kingdom. PMID- 8720276 TI - Subclinical impairment in the median nerve across the carpal tunnel among female VDT operators. AB - Sensory nerve conduction velocities in the palm-to-finger (SCV-pf), wrist-to finger (SCV-wf), wrist-to-palm (SCV-wp), and wrist-to-elbow (SCV-we) segments and the distribution of nerve conduction velocities in the right median nerve were determined among 27 female operators aged 19-37 and 19 healthy women (controls) aged 19-31 to estimate the prevalence of subclinical carpal tunnel syndrome (CTS). Also, the WF/PF ratio, dividing the SCV-wf by the SCV-pf, was calculated to assess abnormalities of nerve conduction within the carpal tunnel. The operators were engaged in data entry in front of a visual display terminal (VDT) for about 6 h/day, and their working duration was between 1 and 17 (mean 6) years. The SCV-wf, SCV-wp, and WF/PF ratio in the operators were significantly lower than those in the controls. The rate of persons with the WF/PF ratio of less than 90%, i.e., below normal limits in the 19 controls, was significantly higher in the operator group (37%) than in the control group (0%). The operators complained of more symptoms related to CTS than did the controls, but any symptoms were not associated with slowing of nerve conduction velocities in the operators. In the controls, the WF/PF ratio was not closely correlated with skin temperature or age despite the presence of significant relations between skin temperature and the SCV-wf, SCV-pf, and SCV-wp; the interpersonal variability of the WF/PF ratio was much smaller than that of all SCVs. In the light of the present and previous studies, the rate of VDT operators with subclinical CTS seems to be high, independent of its symptoms. Also, the WF/PF ratio will be a useful and reliable screening method for the early detection of CTS due to repetitive wrist and finger movements involved in work. PMID- 8720277 TI - Three-year survey of blood lead levels in 8828 Taiwanese adults. AB - The purpose of this study was to investigate environmental lead exposure in the general Taiwanese population. A total of 8828 Taiwanese adults selected by a multistage sampling method were investigated. Characteristics of the participants were ascertained by questionnaire and 10 ml venous blood was drawn by public health nurses. The blood specimens were distributed to six laboratories for blood lead level (BLL) measurement. A quality control program was applied during the analysis of the BLLs in order to improve precision and accuracy. The arithmetic mean BLL of the 8828 Taiwanese adults was 7.70 +/- 5.23 micrograms/dl, with a maximal level of 69.1 micrograms/dl. The median was 6.5 micrograms/dl and the 90th percentile was 14.0 micrograms/dl. After logarithmic transformation, the geometric mean was 1.84 +/- 0.67 microgram/dl. This study also found that elevated BLLs were associated with certain personal characteristics, i.e., gender, ethnic group, and education level; life-style factors, such as smoking, alcohol consumption, Chinese herbal drug consumption, milk consumption, and sources of drinking water; residential location, i.e., level of urbanization; and occupational history of lead exposure. However, age, floor level of residence, distance from house to road, and betel nut consumption were not associated with elevated BLLs. These results showed that BLLs in the Taiwanese population were not higher than those in developed and developing countries. Most of the influencing factors were also found in other studies; however, local factors such as ethnic group, Chinese herbal drug consumption, and sources of drinking water are important considerations in Taiwan when examining ways to prevent overexposure to lead in the general population. PMID- 8720278 TI - Behaviour of urinary 2,5-hexanedione in occupational co-exposure to n-hexane and acetone. AB - We analysed the relationship between free 2,5-hexanedione (2,5-HD) and total 2,5 HD in the urine of 87 workers exposed to n-hexane and other solvents (hexane isomers, acetone and toluene), in relation to different working conditions. The concentration of free 2,5-HD in urine of workers exposed to n-hexane was about 12% of total urinary 2,5-HD. The most significant correlation (r = 0.936) was that of total 2,5-HD in urine with environmental n-hexane and exhaled air. With equal exposure to n-hexane, the concentrations in urine of free and total 2,5-HD increased when cutaneous absorption was involved (gloves not used), during the working week and with co-exposure to acetone. An analysis of the relationship between combined exposure to acetone and urinary concentrations of the various forms of 2,5-HD suggests that acetone might influence the toxicokinetics of n hexane, increasing the proportion of free 2,5-HD. PMID- 8720279 TI - Physiological comparison of three interventions in light assembly work: reduced work pace, increased break allowance and shortened working days. AB - An industrial assembly task known to imply a high risk for shoulder-neck disorders was simulated in the laboratory. Eight females (aged 22-32 years) were trained to manage industrial work pace (120 according to the methods-time measurement system, MTM). They carried out seven work protocols at different days with different combinations of work pace (120 or 100 MTM), break allowance (20 min of active or passive breaks added every 2 h), and duration of the working day (2, 4 or 6 h). During 6 h of work at 120 MTM the electromyographic (EMG) amplitude from the upper trapezius muscle increased by about 11%, the EMG zero crossing rate decreased by about 2.5%, and perceived fatigue increased by about 4 CR10 scale units. When work pace was reduced to 100 MTM, the upper trapezius EMG amplitude decreased by 20% and became less variable. Heart rate decreased by about 10 bpm, perceived fatigue decreased by about 1 CR10 scale units, and shoulder tenderness was reduced by about 5%. However, the work task could still not be performed in a physiological steady state. Added breaks, whether active or passive, had no apparent effects on upper trapezius load during work or on physiological responses. Recovery of EMG, maximal strength, heart rate and blood pressure sensitivity, and tenderness was complete 4 h after work, independent of the preceding work conditions. These findings suggest that a limitation of the daily duration of assembly work may be more effective in limiting acute fatigue than reduced work pace or increased break allowance. PMID- 8720280 TI - Elevated interleukin-8 in the alveolitis of individuals with asbestos exposure. AB - Asbestosis is a fibrotic and inflammatory interstitial lung disease occurring after chronic occupational exposure to asbestos. An alveolitis has been described with activated alveolar macrophages and increased neutrophils as sampled by bronchoalveolar lavage (BAL). Animal models and in vitro studies demonstrate that asbestos can stimulate alveolar macrophages to release neutrophil chemotactic factor. We performed BAL on 18 nonsmoking individuals with asbestos exposure and observed a twofold increase in percent neutrophils recovered. Alveolar macrophages cultured in vitro from the asbestos-exposed individuals spontaneously released significant amounts of the neutrophil chemotaxin, interleukin-8 (IL-8). In addition, the alveolar macrophages expressed a 2.7-fold increase in steady state mRNA levels compared to unexposed normal controls utilizing the reverse transcriptase/polymerase chain reaction. In vitro experiments confirmed that crocidolite or chrysotile asbestos could stimulate the release of IL-8 from mononuclear phagocytes in a dose-dependent fashion. We conclude that asbestos exposure causes a mild neutrophilic alveolitis, and that IL-8 is one potential mediator capable of contributing to this inflammation in the lower respiratory tract. PMID- 8720282 TI - Endogenous and exogenous factors in carcinogenesis: limits to cancer prevention. AB - Organ-specific cancer incidence rates can vary dramatically between low- and high incidence areas. Such differences are due to (1) heritable susceptibility determinants, (2) risk factors associated with the environmental and local living conditions (e.g., viruses, pollution), and (3) personal life-style factors. For organs showing large differences between cancer registries, exogenous factors might be most important, while for organs showing only small differences, endogenous and unavoidable factors are expected to be more important. In this paper, a working hypothesis based on descriptive cancer epidemiology is presented to estimate, in a quantitative manner, the unavoidable contribution to the process of carcinogenesis and to discuss limitations to individual cancer prevention. Cumulative cancer incidence rates for a 75-year period of life (CR74, in percent) were taken from IARC Scientific Publication No. 120 (1992). For each organ, values were ranked in ascending order, and the ratio between high-rate and low-rate registries (90th percentile/10th percentile) was determined. This measure of variability among registries differed strongly between organs. Largest ratios were seen for organs with well-known exogenous risk factors, such as pharynx, lip, tongue, mouth, liver, esophagus, and melanoma in males, and lung, esophagus, gallbladder, liver, and bladder in females. Small ratios were seen for rectum, brain, colon, and Hodgkin's disease in males, and breast, rectum, ovary, brain, and colon in females. It is concluded that the process of carcinogenesis in the latter organs has a stronger endogenous/unavoidable component, for some tissues possibly of hormonal type. A fictitious population was composed where, for each organ, the minimum reported cancer rate was taken. When based on all cancer registries world-wide, CR74 sums over all sites of 2.0% and 2.3% resulted in males and females, respectively. When only Central/Western European countries were included in the analysis in order to reduce differences in risk factors nos. 1 and 2, the sum of the minimum values was 10.4% and 8.7%. After correction of the data for smoking, 'minimum' cancer incidence rates in males and females were estimated to be 7.6% and 6.8%. Based on a median cancer incidence rate for nonsmoking males in Europe of about 21%, therefore, individual preventive measures taken by a nonsmoker can reduce the cancer risk, on average, 'only' by a factor of about 3. A considerable fraction of cases thus appears to be hardly avoidable. PMID- 8720281 TI - Change in digital blood flow with simultaneous reduction in plasma endothelin induced by hand-arm vibration. AB - Involvement of endothelium-derived relaxing factor (EDRF) or endothelium-derived constricting factor (EDCF) has been proposed as the pathophysiologic mechanism of vibration-induced white finger (VWF). Recent evidence that endothelin is a potent vasoconstrictor peptide indicates that it may play a role in vasoregulation during vibration exposure through the local actions of EDRF or EDCF. Therefore, we examined the effects of grasping (50 N) and hand-arm vibration (50 m/s2 rms, 120 Hz, x-axis) on digital blood flow (DBF) and on the level of plasma endothelin in seven healthy male office workers. Grasping decreased DBF without affecting endothelin, and vibration increased DBF with a simultaneous reduction in endothelin. The grasping-induced decrease in DBF seemed to be due to mechanical compression of the vessels. The negative correlation between DBF and endothelin during vibration exposure suggests that a reduction in release of endothelin from smooth muscle into the vessel cavity during vibration leads to vasodilatation, possibly attributable to the local axon reflex. PMID- 8720283 TI - delta-Aminolevulinic acid in plasma or whole blood as a sensitive indicator of lead effects, and its relation to the other heme-related parameters. AB - To evaluate the subclinical effect of lead exposure, we determined delta aminolevulinic acid (ALA) levels in plasma (ALA-P), blood (ALA-B), and urine (ALA U) and the activity of delta-aminolevulinic acid dehydratase (ALAD) in lead workers. Almost all of the ALA molecules in blood were present in plasma and not in blood cells, irrespective of the blood lead concentration (Pb-B). ALA-P or ALA B levels increased slowly at Pb-B levels below 40 micrograms/dl (slow phase) and rapidly at levels above 40 micrograms/dl (rapid phase). In both phases, ALA-P and ALA-B were well correlated with Pb-B and ALAD activity. The threshold value (no effect level) of Pb-B for elevation of the ALA-P or ALA-B level was coincident with that for ALAD inhibition; the value was around 5 micrograms/dl. In the rapid phase, ALA-P increased continuously up to 100 micrograms/dl of Pb-B, while ALAD activity reached a plateau. Receiver operative characteristic (ROC) plot analyses indicated that ALA-P and ALAD activity [ALAD(u)] had a similar diagnostic value at Pb-B levels between 10 and 40 micrograms/dl, although ALAD(%), the remaining ALAD activity as a percentage of the whole activity restored by zinc and dithiothreitol, had the most powerful diagnostic efficiency at these Pb-B levels. By contrast, ALA-U and zinc protoporphyrin were less effective for the diagnosis of lead exposure than ALAD and ALA-P. These findings indicate that ALA-P is the best discriminators of lead exposure form baseline to high levels of exposure. PMID- 8720284 TI - A new risk of occupational disease: allergic asthma and rhinoconjunctivitis in persons working with beneficial arthropods. AB - Allergic asthma and rhinoconjunctivitis have been reported in workers who are constantly in close contact with arthropods during the course of breeding and research. We investigated a group of ten people working with beneficial arthropods in a "bio-factory." The following tests were carried out: skin prick test with allergens present in the workplace, baseline pulmonary function test, aspecific bronchial provocation test with ultrasonically nebulized distilled water, and pulmonary function test after workplace exposure in two subjects. The results demonstrated a good correlation between skin prick test, previous RAST positive reactions, clinical symptoms, and exposure. Pulmonary function test after workplace exposure proved positive in one asthmatic subject and in one subject with rhinoconjunctivitis. It is concluded that the evidence supports the occupational nature of these allergic diseases and that there is a need for careful surveillance and environmental monitoring within the workplace. PMID- 8720285 TI - Introduction: combination antiviral therapy in chronic hepatitis C. PMID- 8720286 TI - Antiviral therapy for chronic hepatitis C viral infection. AB - Hepatitis C affects at least 200 million people worldwide. It can be followed by chronic hepatitis, cirrhosis, and primary liver cancer. Outcome assessments in controlled trials of antiviral therapy are based on serum transaminase values, serum HCV-RNA determinations, and liver biopsy scores. Patients most likely to respond to antiviral treatment are relatively young, have low serum HCV-RNA and transaminase levels, and do not have cirrhosis. Patients whose disease is caused by genotype 1b HCV isolates are unlikely to respond. Interferon alfa (3 million units [MU] three times a week for 6 months) is associated with a 50% response rate and a 50% relapse rate--an overall response rate of 25%. Increasing the duration of therapy may increase the sustained response rate. Ribavirin, given orally, may be used for patients who fail to respond to or relapse after interferon therapy. Its side effects are few. Treatment results in a fall in transaminase levels and some decrease in hepatic inflammation, but serum HCV RNA (viral titer) is unaltered. Combinations of interferon with ribavirin are giving promising results with increased sustained, complete responses. PMID- 8720287 TI - The effect of interferon alfa and ribavirin combination therapy in naive patients with chronic hepatitis C. AB - BACKGROUND: Chronic hepatitis C may lead to cirrhosis and hepatocellular carcinoma in a subset of patients. Because response rates with interferon alfa therapy are unsatisfactory, new therapies are needed. METHODS: We conducted a three-arm, randomized trial in 45 interferon-naive men (mean age 40.6 +/- 12 years) with chronic hepatitis C to compare treatments: group A, ribavirin alone (15 mg/kg daily for 6 months); group B, interferon alone (3 MU thrice weekly for 6 months); and group C, interferon plus ribavirin at the above doses. Histologic outcomes of therapy were assessed by pretreatment and post-treatment liver biopsies. RESULTS: In group A, alanine aminotransferase levels normalized during therapy in 66% of those with HCV-1b and 34% of those with HCV-2a, but all patients relapsed after treatment ended. In group B, alanine aminotransferase levels normalized during treatment in 66%, 75%, and 100% of patients infected with HCV-1b, HCV-2a, and HCV-3, respectively; however, a sustained response was noted in only 25% of those with HCV-3. In group C, a sustained normalization of alanine aminotransferase with negative serum HCV RNA was seen in 20% of those with HCV-1b, 40% of those with HCV-2a, and 75% of those with HCV-3 12 months after therapy. One year after therapy ended, group C demonstrated a significant sustained response (47%) as well as a significant reduction in piecemeal necrosis and portal inflammation (p < 0.05). CONCLUSIONS: Combination therapy was significantly superior to ribavirin or interferon monotherapy in producing a sustained response in interferon-naive patients with chronic hepatitis C (p < 0.05). The results of our study suggest that ribavirin potentiates the effect of interferon therapy in chronic hepatitis C. PMID- 8720288 TI - Combination antiviral therapy with ribavirin and interferon alfa in interferon alfa relapsers and non-responders: Italian experience. AB - BACKGROUND/AIMS: A sustained biochemical and virologic response to standard interferon therapy for chronic hepatitis C is seen in no more than 25% of patients, and the efficacy of re-treatment or of higher doses in non-responders and relapsers has not been established. A more effective therapy for interferon alfa-resistant hepatitis C is needed. METHODS: A study of ribavirin plus interferon alfa combination therapy was conducted in 30 patients with chronic hepatitis C resistant to a previous standard course of interferon alfa (14 interferon non-responders, 16 interferon relapsers). Patients were randomly assigned to receive either ribavirin, 800 mg daily, and interferon alfa, 3 MU thrice weekly (n = 15), or interferon alfa alone, 3 MU thrice weekly (n = 15), for 6 months. RESULTS: At the end of treatment, normal alanine aminotransferase levels were observed in eight patients in the combination therapy group: one (14%) interferon non-responder and seven (87%) interferon relapsers (p = 0.01). Six months post-therapy, sustained normalization of alanine aminotransferase was achieved in seven (87%) interferon alfa relapsers, but not in any of the interferon alfa non-responders (p = 0.001). In the group of patients treated with interferon alfa alone, sustained normalization of alanine aminotransferase was observed in one interferon relapser only. Serum HCV RNA became negative in eight patients receiving combination therapy--two (28%) interferon non-responders and six (75%) interferon relapsers. Six months later, circulating HCV RNA remained negative in seven patients: one (14%) interferon non-responder and six (75%) interferon relapsers (p = 0.04). Sustained clearance of HCV RNA was not observed in patients re-treated with interferon alone. The sustained response to combination therapy was accompanied by reduced hepatic necroinflammatory activity on liver biopsy. Hepatitis C virus genotype was not significantly associated with response to combination therapy. Side effects were mild and well tolerated. CONCLUSIONS: Our experience indicates that combination therapy of ribavirin plus interferon alfa induces sustained biochemical, virologic, and histologic responses in most patients who are interferon relapsers. PMID- 8720290 TI - New approaches to the treatment of hepatitis C virus infection after liver transplantation using ribavirin. AB - BACKGROUND: Chronic liver failure resulting from hepatitis C virus infection often necessitates orthotopic liver transplantation. Recurrence of hepatitis C virus infection after transplantation is inevitable, and the infection is usually severe. Interferon and ribavirin have both been used to treat hepatitis C virus infection after liver transplantation, but neither interferon nor ribavirin monotherapy has demonstrated sustained biochemical or virologic responses or histologic benefit in transplant recipients with recurrent hepatitis C virus infection. The next logical approach to treatment was combination therapy. METHODS: Fourteen patients with recurrent hepatitis C virus infection were treated for 6 months with interferon alfa-2b (3 MU thrice weekly) and oral ribavirin (1000 mg/day). After 6 months, patients were maintained on ribavirin monotherapy until the end of the study. Safety and tolerability were satisfactory, and no patients experienced graft rejection during the study. RESULTS: Alanine aminotransferase levels were normalized in all patients after 6 months of therapy. Serum HCV RNA was negative in nine patients; the other five demonstrated a 50% quantitative reduction of HCV RNA. After a mean follow-up of 18 months, all but one patient maintained normal alanine aminotransferase levels. Twelve out of 14 patients achieved a histologic benefit. CONCLUSIONS: The biochemical and virologic responses and the histologic benefit seen after combination therapy are significantly better than those reported with either interferon or ribavirin monotherapy. Combination therapy appears to be effective in preventing the progression of HCV-related graft disease after liver transplantation. PMID- 8720289 TI - Combination treatment with interferon alfa-2b and ribavirin for chronic hepatitis C in patients who have failed to achieve sustained response to interferon alone: Swedish experience. AB - BACKGROUND: Only 10-20% of patients treated with interferon alfa alone attain long-term benefits. More effective regimens are needed. METHODS: Twenty Swedish patients with chronic hepatitis C virus infection, ten with a prior non-response and ten with a non-sustained response to interferon alfa treatment alone, were treated with interferon alfa-2b and ribavirin in combination for 24 weeks, then followed up for another 24 weeks. Patients received interferon alfa-2b subcutaneously 3 MU thrice weekly and oral ribavirin 1000-1200 mg/day. RESULTS: All ten patients with a prior non-sustained response to interferon alone had a sustained biochemical response with normal aminotransferase levels at follow-up; nine also had a sustained viral response with a negative HCV-RNA test in serum. Among the ten patients with a prior biochemical non-response to interferon alone, five had normal aminotransferase levels at the end of therapy; four were negative for HCV RNA in serum. At follow-up, three had normal aminotransferase levels and a negative HCV-RNA test in serum. No major adverse effect was seen, apart from fatigue and an expected fall in hemoglobin levels from a mean of 155 g/l to 124 g/l at the end of therapy. All patients completed the treatment schedule, but the ribavirin dose was reduced in one patient because of a fall in hemoglobin to 99 g/l. CONCLUSIONS: These results indicate that combination treatment with interferon alfa-2b and ribavirin offers a chance of sustained biochemical response and virus eradication in a subset of patients who fail to achieve sustained response with interferon alfa alone. PMID- 8720291 TI - Hepatitis C virus variants and the role of genotyping. AB - Hepatitis C virus demonstrates considerable divergence in nucleotide sequence. This variation may affect virus detection, disease outcome, and the effectiveness of interferon treatment. For example, infection with genotype 1 is associated with a lower response rate to interferon treatment. Hepatitis C virus can be classified into six distinct types, comprising at least 74 different subtypes. Both types and subtypes are subject to geographical differences in distribution, presumably reflecting the epidemiological history of the virus. However, because this history may be blurred by migration and by commerce in blood products between regions, screening and typing assays must recognize both the major indigenous and more exotic virus genotypes. Little information exists about the role of virus sequence variation in screening for hepatitis C virus antibodies, but there is some evidence that the reactivity of current serological screening assays for hepatitis C virus is genotype dependent. In the future, screening assays may need to include antigens specific for different virus types or may need to be designed with regard to the particular types found in a certain area. Antigenic variation may also mean that an effective vaccine needs to be multivalent to protect against all genotypes present in a given region. There are several polymerase chain reaction-based methods of distinguishing between hepatitis C virus variants. These methods must be continually updated, however, as new sequence variants are discovered. Alternative genotyping assays are based on the host's serological response to virus infection, but these cannot distinguish between virus subtypes and are unsuitable for immunocompromised patients. As more countries are sampled, it is likely that more genotypes will be identified and this may help elucidate the origins of hepatitis C virus. Detailed epidemiological studies may delineate past and current routes of transmission. PMID- 8720292 TI - Future studies of combination therapy for chronic hepatitis C: optimizing response rates for each hepatitis C population. AB - Interferon alfa, the only treatment currently approved for chronic hepatitis C virus infection, has a limited response rate (20-25%) in most patients. Ribavirin has also been used to treat hepatitis C virus infection, but treatment results in only a modest reduction in HCV-RNA levels; most patients relapse after the drug is discontinued. Combination therapy with ribavirin and interferon seems to be a logical approach to the treatment of hepatitis C virus infection. Small studies suggest that combination therapy results in improved sustained response rates compared with either interferon or ribavirin alone. In one study, 6 months of combination therapy was compared with 6 months of ribavirin monotherapy and 6 months of interferon alfa monotherapy. The sustained virologic response rates were 47% for combination therapy, 13% for interferon monotherapy, and 0% for ribavirin monotherapy. This study suggests that combination therapy can potentially double the sustained response rate, but larger studies are needed to evaluate this further. In trial design for clinical studies, many factors must be considered. Determining the response to interferon monotherapy might include establishing liver histology (especially in patients with cirrhosis) and hepatitis C virus genotype--types 1b and 4 are associated with a poor response, which is likely to affect the response to combination therapy. Patients in treatment trials should be followed long term at regular intervals; it is only with prolonged, detailed follow-up that the full effects of antiviral therapy on health and prevention of cirrhosis and hepatocellular carcinoma can be determined. Large multicenter studies are also needed to determine the effect of combination therapy in liver transplant patients. These studies must also address toxicity issues, as interferon may increase the risk of graft rejection post transplant, and the low-grade hemolysis associated with ribavirin may stimulate hepatic iron deposition. PMID- 8720293 TI - [Drug-induced nephrotoxicity]. AB - The incidence of drug-induced nephrotoxicity (DIN) has recently been increasing. Based on developments in molecular biology and cell biology, cultured cells have become a very useful tool for investigating DIN. Especially, the immortalized cell lines derived from well-defined nephron segments are ideal for such studies. As indicators to detect DIN, myosin light chain phosphorylation in glomerular mesangial cells, enzymes of proximal tubule origin such as glycine amidinotransferase, cytosolic free calcium concentration and intracellular ATP content may be useful. The involvement of nitric oxide and heat shock protein in DIN has been reported. Therapeutic effects of growth factors such as HGF or EGF for DIN have been identified. The direct evidence for the involvement of reactive oxygen species and the molecular basis for redox regulation in DIN is required. Cisplatin has been shown to induce apoptosis, and the role of apoptosis in DIN remains to be further clarified. Thus, in parallel with in vivo studies, cultured cells provide an opportunity for clarifying the intracellular mechanism for DIN more precisely and establishing an efficient screening system to develop drugs to prevent DIN. PMID- 8720294 TI - [Molecular and pharmacological studies on signal transduction in the brain]. AB - Many of the extracellular signals such as neurotransmitters and hormones regulate the intracellular concentration of second messengers such as cAMP, cGMP, and calcium ion (Ca2+), diacylglycerol and IP3. Accumulating evidence indicates that protein phosphorylation-dephosphorylation is an important mechanism by which second messengers act to regulate a variety of cellular processes. Ca2+/calmodulin-dependent protein kinase II, cAMP-dependent protein kinase and protein kinase C are three major classes of protein kinases in the central nervous system. In an attempt to elucidate the physiological roles of the protein kinases, I have been studying the substrate proteins and functional significance of the enzymes and phosphorylated proteins. For these purposes, I investigated the phosphorylation-dephosphorylation of cytoskeletal proteins such as microtubule-associated protein 2 and tau, which are involved in the assembly disassembly of microtubules and the production of abnormally phosphorylated forms of tau in neurofibrillary tangles in Alzheimer's disease brain. As the natural consequence, studying the protein phosphatases is significant for elucidating the switch-off mechanism of protein phosphorylation. Thus, I have been investigating the functional significance of protein phosphorylation-dephosphorylation for the elucidation of signal transduction in the brain, which is widely involved in the regulation of brain functions. PMID- 8720295 TI - [Behavioral studies of KSG-504, a new CCK-A receptor antagonist]. AB - The effects of KSG-504 after intravenous administration on behavior and other central functions were studied. KSG-504 did not affect the general behavior of dogs up to the dose of 30 mg/kg, but the drug (100 mg/kg, i.v.) caused vomiting in 3 out of the 5 dogs. Moreover, KSG-504 (1-30 mg/kg, i.v.) had no effects on spontaneous motility, thiopental-induced sleep, acetic acid-induced writhing in mice and satiety in rats. A high dose of CCK-8 (100 micrograms/kg or more) suppressed spontaneous motility, writhing and satiety, and prolonged sleep when administered subcutaneously. The behavioral changes induced by CCK-8 were antagonized by KSG-504 in a dose-dependent manner (1-30 mg/kg, i.v.). When KSG 504 was administered intravenously to rabbits at the dose of 10 mg/kg or 0.5 mg/kg/min for 120 min, we could not detect the drug in the cerebrospinal fluid, indicating that KSG-504 does not cross the blood-brain barrier after peripheral administration of the drug. Thus, the inhibitory effect of KSG-504 on CCK-8 induced behavioral changes may be the result of antagonism at peripheral CCK-A receptors. PMID- 8720296 TI - [Effects of KSG-504, a new CCK-A receptor antagonist, on gallbladder and gastrointestinal functions]. AB - We investigated the interaction of KSG-504 with CCK-8- or pentagastrin-induced gallbladder and gastrointestinal responses in vitro and in vivo. KSG-504 (10(-7) 10(-4) M) inhibited CCK-8-induced contractions of both isolated guinea pig gallbladder and rabbit terminal cavity of the bile duct in a concentration dependent manner. Furthermore, intravenous administration of KSG-504 also dose dependently inhibited CCK-8-induced gallbladder contraction in anesthetized guinea pigs with an IC50 value of 0.23 mg/kg. In conscious mice, KSG-504 inhibited both CCK-8- and egg yolk-stimulated gallbladder emptying in a dose dependent manner (IC50: 13.3 and 9.6 mg/kg, respectively). The CCK-8-induced delay of gastric emptying in conscious rats was also antagonized by KSG-504 with an IC50 value of 3.78 mg/kg, i.v., whereas pentagastrin-stimulated gastric acid secretion in anesthetized rats was not affected by KSG-504 at all. KSG-504 (1 mg/kg, i.v.) also inhibited CCK-8-induced duodenal and jejunal contractions in anesthetized rabbits. These results indicate that KSG-504 exerts an antagonistic effect on CCK-A receptors in the gastrointestinal tract, but not on gastrin receptors in the stomach. PMID- 8720297 TI - Bicyclic dioxolanes as potential antimuscarinic agents. AB - A series of rigid compounds--derivatives of 2,2-diphenyl-[1,3]-dioxolan-4 ylmethyl-dimethylamine methiodide (1b)--has been synthesized and tested to evaluate affinity and selectivity for M1, M2, and M3 muscarinic receptors. The stereochemistry of the annulation does not influence the activity; the optimal distance between nitrogen and the benzhydryl group seems to be that with the nitrogen in the rigid ring (2b) or the nitrogen directly bound at the cyclopentane nucleus (9b, 11b). The different structure-activity relationships between tertiary amines (a series) and the corresponding quartnerary salts (b series) suggest a different binding to the receptor sites. PMID- 8720298 TI - Pharmacokinetics of the new thyrotropin releasing hormone analogue montirelin hydrate. 1st communication: plasma concentrations, metabolism and excretion after a single intravenous administration to rats, dogs and monkeys. AB - The plasma concentrations, metabolism and excretion were studied in rats, dogs and monkeys after a single intravenous administration of 14C-montirelin hydrate (CAS 90243-66-6, 14C-NS-3). 1. The plasma concentration of radioactivity and the area under curve (AUC0-infinity) increased with the dose after intravenous administration of 0.25, 1 and 4 mg/kg to male rats. 2. Until 4 h after intravenous administration of 0.25 mg/kg, the plasma concentration of radioactivity decreased with half-lives of 0.512, 0.771 and 0.786 h in male rats, dogs and monkeys, respectively. The plasma concentration of radioactivity in rats increased 6-10 h after administration, suggesting enterohepatic circulation. 3. The plasma concentration of NS-3 in various animal species decreased biphasically after intravenous administration. The elimination half-lives, t1/2 beta, in rats, dogs and monkeys were 0.324, 0.679 and 0.682 h, respectively, and the steady state volumes of distribution, Vdss, were 0.248, 0.319 and 0.306 l/kg, respectively. 4. The binding of NS-3 to serum protein was less than 3% in rats, dogs, monkeys and humans. 5. Within 48 h after intravenous administration of 1 mg/kg to male rats, the excretion of radioactivity in urine, feces and expired air was 67.3, 15.0 and 14.8% of the dose, respectively (total 97.1%). The biliary excretion of radioactivity was 26.6% within 24 h after dosing. 6. No sex-related difference was found in plasma concentration of radioactivity or the excretion of radioactivity in urine, feces and expired air after intravenous administration of 1 mg/kg to male and female rats. 7. In male dogs, the excretion of radioactivity in urine and feces was 92.3 and 5.6% of the dose, respectively, during 72 h after intravenous administration of 0.25 mg/kg (total 97.9%). 8. In male monkeys, the excretion of radioactivity in urine and feces was 91.7 and 2.9% of the dose, respectively, during 72 h after intravenous administration of 0.25 mg/kg (total 94.6%). 9. The urinary excretion of NS-3 and its main metabolite CNK-6004 (deamidation product) during 24 h after intravenous administration was 31.4 and 25.3% of the dose in male rats, 45.4 and 30.1% in male dogs, and 20.2 and 48.0% in male monkeys, respectively. The biliary excretion of NS-3 and CNK-6004 in male rats was 1.1 and 17.4%, respectively, during 24 h after administration. 10. The radioactivity in the bile collected from donor rats that had received 14C-NS-3 was easily absorbed from the intestinal tract of recipient rats after intraduodenal administration of the donor's bile. PMID- 8720299 TI - Pharmacokinetics of the new thyrotropin releasing hormone analogue montirelin hydrate. 2nd communication: distribution and transfer into the fetus and milk after a single intravenous administration and pharmacokinetics and enzyme induction after repeated intravenous administration to rats. AB - Pharmacokinetics of 14C-labeled montirelin hydrate (CAS 90243-66-6, NS-3) in rats was studied after single or repeated intravenous administration. 1. Radioactivity concentrations in tissues after single administration to male and female rats were highest in the kidney followed (in this order) by plasma, liver, blood, pancreas, uterus (female rats), lung and skin and low in various brain sites 5 min after administration. The concentrations in most tissues were practically parallel to those in plasma over the 24-h period after administration. After decreasing rapidly the concentrations rose slightly for 10 h and then decreased gradually. 2. Five min after single administration to male rats, the concentration of the main metabolite CNK-6004 (deamidated product) was lower in the plasma, but higher in the liver and kidney than the NS-3 concentration. From 0.5 to 2 h after administration, the concentration of CNK-6004 was higher than that of NS-3 in the plasma, liver and kidney, accounting for 33-64% of the radioactivity concentration. 3. After administration to rats on the 18th day of pregnancy, the radioactivity concentrations in the fetal whole body and fetal tissues peaked later than those in the maternal plasma, tissues and placenta. The maximum concentration in the fetal tissues was 2% or less of that in the maternal plasma. 4. After administration to lactating rats, the radioactivity concentration in milk reached the maximum 10 h after administration, and decreased gradually in parallel with the concentration in the plasma 24 to 168 h after administration. 5. During repeated once daily administration for 10 days, the radioactivity concentration in the plasma 24 h after each administration reached practically steady state level after the 7th administration and decreased with a half-life of 38.1 h after the last administration. 6. The radioactivity concentrations in most tissues after the last administration were not significantly different from those after a single administration. Decreased elimination of the radioactivity was observed in the white fat and skin, in which radioactivity levels were higher than those in the other tissues a long time after the last dose. 7. Excretion of the radioactivity in the urine and feces during repeated administration was constant after the 2nd administration. The excretion by 168 h after the last administration was 66.9 and 14.3% of the cumulative dose in the urine and feces, respectively (total: 81.2%). 8. The composition of metabolites in the plasma, liver, kidney and urine after the last administration did not differ markedly from that after a single administration. Once daily repeated administration for 7 days had no effect on the liver drug metabolizing enzyme activities. PMID- 8720300 TI - Pharmacokinetics of the new thyrotropin releasing hormone analogue montirelin hydrate. 3rd communication: identification of metabolites in rat urine. AB - The metabolism of montirelin hydrate (CAS 90243-66-6, NS-3) was studied in rats after intravenous administration of 14C-labeled or unlabeled NS-3. 1. Four radioactive metabolites (M-1 to M-4) were found in the urine after administration of 14C-NS-3. M-3 (major metabolite) and M-2 showed the same Rf values as (-)-N [[(3R,6R)-6-methyl-5-oxo-3-thiomorpholinyl]carbonyl]-L-histi dyl-L- proline (CNK 6004) and (+)-N-[[(3R,6R)-6-methyl-5-oxo-3- thiomorpholinyl]carbonyl]-L-histidine (CNK-6001), respectively. 2. M-3 and M-2 were purified from the urine after administration of unlabeled NS-3, and their chemical structures were identified by mass spectrometry, 1H-nuclear magnetic resonance spectroscopy, thin-layer chromatography and high performance liquid chromatography. Consequently, M-3 was identified as CNK-6004 formed by deamidation at a prolinamide moiety of NS-3, and M-2 as CNK-6001 formed by deprolination of CNK-6004. PMID- 8720301 TI - Electromechanical effects of newly synthetized propafenone derivatives on isolated guinea-pig heart muscle preparations. AB - Inotropic, chronotropic and beta-adrenoceptor blocking activities of the newly synthetized propafenone derivatives 1-(2-(3-diethylamino-2-hydroxypropoxy)phenyl 3-phenyl-1-propanone hydrochloride (AM 03), 1-(2-(2-hydroxy-3-(1 piperidyl)propoxy)phenyl)-3-phenyl-1-propanone hydrochloride (AM 05), N,N dimethyl-N-(2-hydroxy-3-(2-(3- phenylpropionyl)phenoxy)propyl)-propylammonium iodide (TH 41), N,N- diethyl-N-(2-hydroxy-3-(2-(3-phenylpropionyl)phenoxy)propyl) methylammon ium iodide (AM 07), and N,N-diiso-propyl-N-(2-hydroxy-3-(2-(3 phenylpropionyl)phenoxy)prop yl)- methylammonium iodide (AM 09) studied in isolated, electrically stimulated papillary muscles and spontaneously beating right atria of guinea pigs. In comparison with propafenone the tertiary amines AM 03 and AM 05 showed a higher negative inotropic potency, while the quarternary amines TH 41, AM 07 and AM 09 were less effective. With regard to their negative chronotropic action, AM 03 and AM 05 were more and TH 41, AM 07 and AM 09 less potent than the parent drug. In contrast to propafenone none of its derivatives exerted beta-adrenoceptor blockade at the concentrations studied. PMID- 8720302 TI - Investigation of the positive inotropic action of 6,7-dimethoxy-3-cyano-3,4 dihydrocarbostyril. AB - 6.7-Dimethoxy-3-cyano-3.4-dihydrocarbostyril (7A61, 1), a 2-quinolinone derivative, showed interesting cardiotonic properties in vitro. It exerted potent positive inotropic effects, without any change in heart rate, in guinea pig atrial preparations, and its effects were not dependent upon inhibition of phosphodiesterases, opening of Na+ channels, or inhibition of adenosine receptors. Notwithstanding its structural relationships with milrinone, vesnarinone, and other quinolinones endowed with cardiotonic activity, it appeared that 7A61 does not share any of the main mechanisms of action involved in the pharmacological activity of these compounds. Other experiments also excluded the involvement of other well known mechanisms of cardiac stimulation, such as adrenergic receptor activation or opening of Ca++ channels, the mode of action of 7A61 remaining unexplained. However, contrary to milrinone, 7A61 did not exert cardiotonic effects in human ventricular muscle preparations, thus discouraging further investigation. Although the lack of effect on human heart could not be predicted from the results obtained in several cardiac preparations commonly used in laboratory investigations, the scarce efficacy of 7A61 in comparison with milrinone in dog and cat papillary muscle and in guinea pig ventricular muscle suggests the utility of the latter preparations in laboratory experiments aimed at selecting new potential cardiotonic agents. PMID- 8720303 TI - Antihypertensive effects of lercanidipine in experimental hypertensive rats and dogs. AB - The antihypertensive action of lercanidipine (CAS 132866-11-6, Rec 15/2375), a new 1,4-dihydropyridine (1,4-DHP) calcium entry blocker (CEB), was examined in spontaneously hypertensive rats (SHR) and renal hypertensive dogs after acute and repeated administration, in comparison to several reference 1,4-DHPs. In acute experiments in SHR, lercanidipine reduced diastolic blood pressure showing a potency similar to felodipine and 2-3 fold higher than those of nicardipine and nitrendipine, after both intravenous and oral administration. Analysis of the area under the curves of percent reduction of diastolic blood pressure exerted for 3 and 8 h after intravenous and oral administrations, respectively, showed that the duration of the antihypertensive effect of lercanidipine was longer than that of the reference dihydropyridines. After repeated administrations to SHR no tachyphylaxis was observed, as indicated by the marked and persistent decrease in systolic blood pressure elicited by lercanidipine, given orally once a day for 21 days. Moreover, starting from the first week of treatment, the daily basal values of systolic blood pressure of the rats treated with lercanidipine were significantly lower than those of the placebo-treated group. In renal hypertensive dogs, after acute oral administration, lercanidipine was as potent as nitrendipine. After repeated administration, the action of lercanidipine was longer lasting than that of nicardipine and no decrease in the antihypertensive effects was observed. The in vivo studies show that lercanidipine has a potent and long-lasting antihypertensive profile, suggesting that this compound may be used for once-a-day treatment. PMID- 8720305 TI - Cardiotoxic and drug interaction profile of the second generation antihistamines ebastine and terfenadine in an experimental animal model of torsade de pointes. AB - Second generation antihistamines are widely used because of their efficacy in treating allergic disorders without significant sedative side effects. Recent clinical evidence shows that some of the early prototypes in this class, namely terfenadine and astemizole, have the potential for producing torsade de pointes, a rare form of ventricular arrhythmia that is life-threatening. Important questions have been raised as to whether this is a property shared by newer, recently-introduced second generation antihistamines. The objective of this study was to characterize and compare the ECG and cardiovascular effects of terfenadine (CAS 50679-08-8) and ebastine (CAS 90729-43-4), a new second generation antihistamine, in an experimental animal model predictive of the cardiotoxic proclivity of these agents. Also, the drug interaction effect of the antifungal drug ketoconazole (CAS 65277-42-1) was evaluated, which blocks hepatic first-pass biotransformation of ebastine and terfenadine leading to increased cardiotoxity of terfenadine in man, on the ECG effects of terfenadine and ebastine in this animal model. Terfenadine (10 mg/kg) and ebastine (50 mg/kg) were administered intravenously to anesthetized guinea pigs. Electrocardiographic (ECG) and cardiovascular parameters (blood pressure and heart rate) were measured during the course of the experiment. The ECG wave form was analyzed to determine QTc interval, PR interval, QRS interval and heart rate. In separate studies in conscious guinea pigs, the effect of oral ketoconazole (200 mg) on the ECG effects of oral terfenadine (60 mg) and ebastine (10 mg) was studied. Terfenadine (10 mg/kg) and ebastine (50 mg/kg) produced significant prolongation of the QTc interval and disruption of the ECG signal when given intravenously to anesthetized guinea pigs. The ECG effects were characterized by large amplitude, morphologically aberrant T-waves, and instances of arrhythmogenic activity. Both drugs produced pronounced bradycardia and hypotension. In conscious animals, pretreatment with oral ketoconazole significantly enhanced the QTc interval prolongation effects of terfenadine and ebastine. Oral terfenadine and ebastine, when given alone at the doses tested, were devoid of adverse QTc interval prolongation effects in the conscious guinea pig. In separate studies in conscious guinea pigs, oral loratadine (10 mg; CAS 79794-75-5) given alone or in animals pretreated with ketoconazole did not affect ECG parameters. The present studies show that terfenadine and ebastine share similar cardiotoxic properties characterized by QTc interval prolongation, bradycardia, hypotension and proarrhythmogenic activity in the anesthetized guinea pig. In addition, pretreatment with ketoconazole enhances the QTc interval effect of both drugs, most likely due to the accumulation of parent compound that occurs after blockade of hepatic metabolism by CYP3A4. In conclusion, our findings indicate that ebastine and terfenadine display similarities in their inherent potential for cardiotoxic and adverse drug interaction effects. In contrast, loratadine is devoid of adverse ECG and drug interaction effects. PMID- 8720304 TI - Comparative analysis of the cardiotoxicity proclivities of second generation antihistamines in an experimental model predictive of adverse clinical ECG effects. AB - Terfenadine and astemizole belong to the second generation histamine H1 antagonists and are widely prescribed for allergic and upper respiratory diseases. The popularity of the newer H1 antihistamines is due to their ability to provide relief from allergic symptoms without the undesirable side effect of sedation commonly associated with first generation H1 receptor antagonists such as diphenhydramine and promethazine. Recent clinical evidence that the second generation histamine H1 antagonists terfenadine and astemizole have the potential for inducing life threatening ventricular arrhythmias has raised questions as to whether other drugs in this class have similar cardiotoxic potential. The objective of this study was to evaluate and compare the arrhythmogenic potential of a series of second generation antihistamines in a quantitative experimental model predictive of adverse ECG effects in man. Antihistamines were given intravenously and electrocardiographic (ECG) and cardiovascular parameters (blood pressure and heart rate) were measured. The ECG wave form was analyzed to determine QTc interval, PR interval, QRS interval and heart rate. To determine the relative cardiotoxic potential of the antihistamines, the lowest dose producing significant prolongation of the QTc interval was compared with the dose required to inhibit by 50% the peripheral bronchospasm elicited by histamine at 10 micrograms/kg i.v. (antihistamine ED50). The second generation antihistamines studied were astemizole (CAS 68844-77-9), carebastine (CAS 90729-42-3), cetirizine hydrochloride (CAS 83881-52-1), ebastine (CAS 90729-43-4), norastemizole (CAS 75970-99-9), terfenadine (CAS 50679-08-8) and terfenadine carboxylate (CAS 83799-24-0). The second generation antihistamines astemizole, ebastine and terfenadine produced pronounced dose-dependent QTc interval prolongation effects. These arrhythmogenic effects occurred at doses that were between 1 and 4 times their respective peripheral antihistamine doses. These drugs produced significant disruption of the ECG wave form including large amplitude, morphologically aberrant T-waves and, in some cases, torsades de pointes-type arrhythmias. In contrast, terfenadine carboxylate (100 mg/kg i.v.), norastemizole (20 mg/kg, i.v.) and carebastine (50 mg/kg, i.v.), the major metabolites of terfenadine, astemizole and ebastine, were largely devoid of adverse ECG effects. Similarly, cetirizine (20 mg/kg, i.v.) was also found to not alter ECG or cardiovascular function. These findings demonstrate that terfenadine, astemizole and ebastine exhibit significant arrhythmogenic effects including QTc interval prolongation, bradycardia and distortion of the ECG morphology in the guinea pig. The relative cardiotoxicity of these antihistamines based on the separation of antihistamine activity and adverse ECG effects was similar for terfenadine and astemizole, but slightly less for ebastine. In this model carebastine, cetirizine, loratadine, norastemizole and terfenadine carboxylate are devoid of QTc prolongation effects. Given the structural similarity of terfenadine and ebastine it is not surprising that these drugs produce significant cardiotoxicity in this animal model. Taken together, these results indicate that the ability to cause QTc interval prolongation and the proclivity for producing arrhythmias is not a class effect and is seen only with some second generation nonsedating antihistamines. PMID- 8720306 TI - Influence of non-steroidal anti-inflammatory drugs on the binding kinetics of dansylsarcosine to human serum albumin. Stereoselectivity, steric and inductive effects. AB - The effect of a series of non-steroidal anti-inflammatory drugs (NSAIDs) on the binding kinetics of dansylsarcosine (CAS 72517-44-3, DS), a marker ligand for the benzodiazepine binding site, and human serum albumin (HSA) was studied using the stopped-flow method. Both native (7% glycated) and 25% glycated HSA were used. The binding parameters were determined on the basis of the consecutive model. The DS association rate constant (k2) was 649 +/- 84 s-1 and 375 +/- 13 s-1 for 7% and 25% glycated HSA, respectively. These values were substantially influenced by addition of NSAIDs (molar ratio HSA:NSAID = 2:1), depending on the structure of NSAIDs. The calculated DS dissociation rate constant (k-2) was approximately 20 s 1. this value did not show marked dependence on the degree of glycation or on the presence of NSAIDs at the concentration used. The values were similar to estimates of kd (the displacement rate constant of DS) with the exception of diclofenac (CAS 15307-86-5) where kd was significantly lower, reaching 4.8 +/- 0.4 s-1 and 4.8 +/- 0.6 s-1 vs. k-2 parameters of 14 +/- 2.8 s-1 and 15 +/- 3.7 s 1 for 7% and 25% glycated HSA, respectively. A comparison of the enantiomers R- and S-ibuprofen (CAS 15687-27-1) and the regioisomers fenbufen (CAS 36330-85-5) and ketoprofen (CAS 22071-15-4) showed slight or no stereoslectivity of effects on the DS binding kinetics. However, the binding was influenced by bulk and nature of substituents at the aryl rest of propionic acid. The results obtained for mefenamic acid (CAS 61-68-7) suggest that this NSAID binds to a site of human serum albumin other than site II. Increased concentrations of glycoalbumin, as observed in diabetic patients, are not presumed to have inhibitory effects additional to that of NSAIDs which interact differentially with drugs at site II of HSA. PMID- 8720307 TI - Free radical scavenging activity of curcuminoids. AB - Three natural curcuminoids (curcumin (CAS 458-37-7), demethoxycurcumin, bisdemethoxycurcumin) and acetylcurcumin were compared for their ability to scavenge superoxide radicals and to interact with 1,1-diphenyl-2-picryl-hydrazyl (DPPH) stable free radicals. The results showed that curcumin is the most potent scavenger of superoxide radicals followed by demethoxycurcumin and bisdemethoxycurcumin. Acetylcurcumin was inactive. Interaction with DPPH showed a similar activity profile. The study indicates that the phenolic group is essential for the free radical scavenging activity and presence of methoxy group further increases the activity. PMID- 8720309 TI - Comparative pharmacokinetic evaluation of compressed suppositories of diclofenac sodium in humans. AB - Diclofenac sodium (CAS 15307-79-6) suppositories were formulated using polyethylene glycol 4000 as the base by dispersing the drug in the molten base, congealing the mass, followed by pulverization, sieving and subsequent compression of the resultant granules. These suppositories were evaluated with respect to their pharmacokinetic behaviour in 12 healthy, male human volunteers. The results were compared with those obtained after oral administration of a commercial enteric coated tablet. Bioequivalence between rectal suppositories and commercial tablets was observed with respect to AUC0-infinity and Cmax. However, tmax differed significantly (p < 0.05) in case of rectal administration (0.625 +/ 0.065 h) compared to oral tablet (1.58 +/- 0.06 h). The relative rectal bioavailability was 107.19 +/- 3.2. PMID- 8720308 TI - Antipyretic effect of Lumbricus spencer in acetylsalicylic acid-induced asthma. AB - Lumbricus spencer, which has been used as an antipyretic in Chinese and Japanese folk medicine and whose antipyretic components are identified to be eicosatetraenoic acid and eicosapentaenoic acid, was given orally in a single dose of 500 mg to febrile patients with acetylsalicylic acid-induced asthma (AIA) in placebo-controlled design. The antipyretic effect of Lumbricus spencer was observed with no exacerbation of asthmatic symptoms. Single oral administration of 500 mg of Lumbricus spencer did not affect the respiratory functions in the patients. Moreover, there were no significant changes in symptom scores and airway lability as measured by daily variations in peak expiratory flow after one week administration of 1500 mg/d of Lumbricus spencer. This pilot study suggests that Lumbricus spencer can be administered safely to febrile patients with AIA without bronchospasm. PMID- 8720310 TI - Effects of the novel histamine H2-receptor antagonist (+/-)-(E)-1-[2-hydroxy-2-(4 hydroxyphenyl)ethyl]-3-[2-[[[5- (methylamino)methyl-2-furyl]methyl]thio]ethyl]-2 (methylsulfonyl) guanidine on gastric secretion and gastroduodenal ulcers in rats. AB - The effects of (+/-)-(E)-1-[2-hydroxy-2-(4-hydroxyphenyl)ethyl]-3-[2- [[[5 (methylamino)methyl-2-furyl] methyl]thio]ethyl]-2-(methylsulfonyl)guanidine (CAS 140695-21-2, T-593), a new histamine H2-receptor antagonist, on gastric secretion and experimental gastric and duodenal lesion/ulcer models in rats were examined. The drug administered orally or intraduodenally significantly and dose dependently inhibited both basal and histamine-stimulated acid secretion. Pepsin output was also inhibited by the drug nearly dose-dependently. The acid inhibitory effect of T-593 persisted for 12 h after a single oral administration. T-593 potently protected the gastric mucosa against water-immersion stress-, indometacin- and HCl.acetylsalicylic acid-induced lesions, but it had no effect on HCl.ethanol-induced lesions. T-593 significantly prevented the development of mepirizole-induced duodenal ulcers. Spontaneous healing of kissing gastric ulcers was significantly enhanced when T-593 was administered for 14 days. The antisecretory and antilesion/antiulcer effects of T-593 were similar to those of ranitidine and omeprazole. It is concluded that T-593 is a potent antisecretory and antiulcer drug. PMID- 8720311 TI - General pharmacology of the new non-xanthine adenosine A1 receptor antagonist (+) (R)-[(E)-3-(2-phenylpyrazolo[1,5-a]pyridin-3-yl)acryloyl]-2- piperidine ethanol. AB - FK 453 ((+)-(R)-[(E)-3-(2-phenylpyrazolo[1,5-a]pyridin-3-yl) acryloyl]-2 piperidine ethanol, CAS 121524-18-3) is a potent non-xanthine adenosine A1 receptor antagonist with diuretic and renal vasodilatory activity. The general pharmacology of FK 453 was investigated in mice, rats, guinea-pigs and dogs. In in vivo tests, FK 453 had little effect on the central nervous system (general behaviour, spontaneous motor activity, potentiation of barbiturate anesthesia, anticonvulsant activity, analgesic activity and body temperature), hematological system (bleeding time, coagulation time and recalcification time) and intestinal charcoal transit. FK 453 also did not show any cardiovascular (blood pressure, heart rate and femoral blood flow) or respiratory effects. In in vitro tests, although FK 453 had little effect on noradrenaline-induced contraction in rat vas deferens and histamine-induced contraction in guinea-pig trachea, FK 453 inhibited the acetylcholine-, histamine- and barium-induced contraction in isolated guinea-pig ileum and serotonin-induced contraction in isolated rat stomach. FK 453 also exerted significant inhibitory activity on collagen- and U 46619-induced platelet aggregation. However these effects of FK 453 on isolated tissue and platelet were observed only at high concentrations. These results suggest that FK 453 possesses a selective pharmacological profile, and one promising therapeutic site for this drug is in the kidney. PMID- 8720312 TI - Studies on the effect of the new non-steroidal aromatase inhibitor fadrozole hydrochloride in an endometriosis model in rats. AB - The therapeutic effect of the new nonsteroidal aromatase inhibitor fadrozole hydrochloride (4-(5,6,7,8-tetrahydro-imidazo[1,5a]pyridin-5-yl)benzonitrile monohydrochloride, fadrozole, CAS 102676-31-3, CGS16949A) was assessed using a surgically induced endometriosis model in rats. In nontreated rats, the endometrial transplants on the abdominal wall developed large cysts with fluid. In the fadrozole treated group, the cystic volume of the transplants decreased in a dose-dependent manner. The weights of the hemi-uteri were markedly reduced by fadrozole treatment. Fadrozole produced a dose-dependent increase in percentage of vaginal diestrous days. In ovariectomized group, the growth of the transplants was also suppressed, and the weights of the hemi-uteri decreased markedly. Histologically, the signs of growth suppression and/or atrophic changes such as minimization of luminal size, cuboidal piknotic epithelium and/or contraction of stroma were observed in ovariectomized or fadrozole treated groups. The right uterine horns also showed marked atrophic changes as with the transplants. The present study strongly indicates that the new selective aromatase inhibitor fadrozole should be useful for the treatment of endometriosis. PMID- 8720313 TI - Detection of apoptosis in KG-1a leukemic cells treated with investigational drugs. AB - Four investigational drugs, p-benzoquinone, primine, miconidine acetate, and artesunate (dihydroqinghaosusuccinate), with growth inhibitory activity against flagellatae (e.g. trypanosoma, leptomonas, plasmodium) were investigated for their capability to induce programmed cell death (apoptosis) in human KG-1a leukemic cells. The results were compared with those of three well established cytostatic agents (cisplatin, daunorubicin, cytosine-arabinoside) and ionizing radiation. The antitumor activity of the drugs was validated by a cellular growth inhibition assay. The depletion of glutathione by these four investigational drugs favours the hypothesis that formation of free radicals and subsequent DNA strand breaks may be critical mechanisms of action and that the glutathione redox cycle is involved in detoxification of these reactive molecules. PMID- 8720314 TI - Effects of the new selective protein kinase C inhibitor 4'-N-benzoyl staurosporine on cell cycle distribution and growth inhibition in human small cell lung cancer cells. AB - CGP 41251 (4'-N-benzoyl staurosporine, CAS 120685-11-2) exerts increased selectivity for Ca(2+)- and phospholipid-dependent protein kinase C inhibition. In this study, the effects of CGP 41251 on cell cycle distribution and growth inhibition were examined in SBC3 human small cell lung cancer (SCLC) cell line. CGP 41251 caused the inhibition of cell proliferation and at 1.0 mumol/l showed almost complete effect. In early S phase synchronized SBC3 cells, CGP 41251 at 1.0 mumol/l did not inhibit an initial progression from early S to G2 phase, but it blocked a process from G2/M to G1 phase completely. After removal of nocodazole block, CGP 41251 at 1.0 mumol/l caused DNA re-replication and induction of polyploidy. In nude mice xenograft, CGP 41251 at a dose of 200 mg/kg showed statistically significant inhibition against this tumor with a T/C value of 21.4%. Histopathologically, expansion of central necrosis was observed by the administration of CGP 41251. These results in SBC3 cells indicated that CGP 41251 showed antitumor activity through the inhibition of cell cycle progression from G2/M to G1 phase, and through induction of cells with higher DNA content. PMID- 8720315 TI - Synthesis of ethanone derived from 2-benzoxazolinones and their antimicrobial activities. AB - A number of 3-(4-substituted benzoyl methyl)-2-benzoxazolinones have been synthesized by reacting with 2-benzoxazolinone and 4-substitutet phenacyl bromide in ethanol. Their structures were confirmed by microanalysis, IR and NMR spectral analysis. Possible antimicrobial activity of the compounds was investigated by tube dilution and paper disc techniques using bacteria (Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 25923, Pseudomonas aeruginosa ATCC 27853, Staphylococcus faecalis RSKK 10541) and yeast-like fungi (Candida parapsilosis, Candida albicans, Candida pseudotropicalis, Candida stellaoidea). Among the compounds tested 3-(4-bromo benzoylmethyl)-5-chloro-2-benzoxazolinone (compound 4) and 3-(4-nitro benzoyl methyl)-5-chloro-2-benzoxazolinone (compound 6) showed the most favorable activity. PMID- 8720316 TI - Synthesis and antischistosomal activity of a new 2-nicotinoyl dipeptide analogue of praziquantel. AB - As an approach to a structure-antischistosomal activity-relationship with possible pharmacological potentiation of the anthelmintic drug praziquantel, a new dipeptide analogue, namely, N alpha-nicotinoyl-L-aspartyl-beta-(1,2,3,6,7,11b hexahydro-4H-pyrazino[2- 1a]isoquinoline-4-one)-L-phenylalanine methyl ester, was synthesized and antischistosomally investigated in mice infected with S. mansoni cercariae. Parallely, its simple 2-nicotinoyl analogue was synthesized and tested. Both compounds were less, but still, interestingly active (approximately 63 and approximately 66%, respectively) compared to praziquantel (approximately 90%). PMID- 8720317 TI - In vitro activity of a new rifamycin derivative against Mycobacterium leprae. AB - The antimicrobial effects of T9 (3-(4-cinamyl-1-piperazinyl)iminomethyl rifamycin SV) alone and in combination with ofloxacin, against strains of M. leprae were evaluated, using an in vitro cell-free culture system. The minimum inhibitory concentrations (MICs) of T9 against rifampin-sensitive and rifampin-resistant strains of M. leprae were 0.1 microgram/ml and 0.4 microgram/ml, respectively. Furthermore, in common with rifabutin, but not with rifamycin, T9 demonstrated synergy with ofloxacin against both rifampin-sensitive rifampin-resistant strains of M. leprae. The results suggest that T9, in combination with ofloxacin as part of multidrug regimens, warrants further evaluation as treatment for patients with leprosy. PMID- 8720318 TI - Influence of the antibiotics erythromycin and azithromycin on the pharmacokinetics and pharmacodynamics of midazolam. AB - The pharmacokinetic and pharmacodynamic interaction between azithromycin (CAS 83905-01-5), an azalide antibiotic, and midazolam (CAS 59467-70-8), a short acting hypnotic agent, was investigated in an open, three-way cross-over study, including erythromycin (CAS 114-07-8) as a positive control. Twelve healthy male and female subjects had standard doses of azithromycin (500 mg o.d. over 3 days), or erythromycin (500 mg t.i.d. over 5 days), or no pretreatment. On the day of the last dose, they ingested 15 mg midazolam. Blood samples were collected and psychometric tests performed. Erythromycin pretreatment (E) significantly changed the pharmacokinetics of midazolam compared to control (C), whereas azithromycin (A) had no such effect. The parameters are summarized as follows: area under the concentration-time curve, AUC (C) 173.8 h.ng.ml-1 vs. (E) 662.7 h.ng.ml-1*+ and (A) 220.0 h.ng.ml-1; concentration maxima (C) 67.2 ng.ml-1 vs. (E) 182.3 ng.ml 1*+ and (A) 86.7 ng.ml-1; elimination half-life (C) 2.21 h vs. (E) 4.85 h* and (A) 2.41 h (* p < 0.05 vs. (C), +p < 0.05 vs. (A)). Pharmacodynamic tests (digit symbol substitution test; critical flicker fusion test; subjective analog scale for rating of alertness; duration of sleep) consistently showed significant differences after erythromycin pretreatment compared to control, but not after azithromycin. Erythromycin, but not azithromycin, causes clinically significant changes in the pharmacokinetics and pharmacodynamics of midazolam. PMID- 8720320 TI - The use of structure generators in predictive pharmacology and toxicology. AB - The analysis of pharmacological effects of chemical compounds often requires expensive of database searches. But in many cases, some of the most useful (or toxic) substances are overlooked because they are missing in the experiment schedule or in the database etc. In the recent years quantitative structure activity relationship (QSAR) studies have shown several correlations of physiochemical properties and topological information indices. In the present paper, the use of an automatic structure generator is studied for detecting compounds with maximal activity by (carefully) extrapolating these correlations. So assumptions of optimal activity can be easier evidenced or discarded. The ideas are outlined on barbiturates and monoketones. PMID- 8720319 TI - Wound healing effect of the new imidazole antimycotic lanoconazole in rats. AB - The wound healing effect of cream preparations of lanoconazole (CAS 101530-10-3, NND-318), an antimycotic imidazole compound, was examined using an excisional open skin-wound model produced on the back of rats. The rapid decrease in the size of wounded areas showed that 0.5% and 1% lanoconazole creams accelerated spontaneous healing. The effectiveness was almost similar to that of an ointment containing 5% deproteinized calf blood extract (DCBE), a wound healing agent on the market. In contrast, neither 1% clotrimazole cream nor 1% bifonazole cream, both of which are imidazole antimycotics, showed an accelerative effect. The wound healing effect of lanoconazole was further confirmed by histological evaluation; a thicker and more cellular granulation tissue was formed, and epidermal regeneration was more stimulated by lanoconazole than by non-treatment or vehicle alone. The effect of lanoconazole on the formation of granulation tissue in rats was also studied using a cotton pellet implantation method. Lanoconazole accelerated the formation of this tissue in terms of dry weight in a dose dependent manner (0.5-4 mg/pellet), and collagen content and angiogenesis also increased in the stimulated tissue, indicating that these accompany the compound-induced acceleration of tissue formation. These results suggest that lanoconazole has a distinctive wound healing effect which is a feature no other imidazole antimycotic is known to possess to date. PMID- 8720322 TI - [Molecular basis for detection of infectious agents]. AB - Elimination of foreign pathogens requires detection of the presence of such microorganisms somewhere in the body. This task relies on specialized cells, among which specific lymphocytes permanently circulate throughout the body searching for signals indicative of the presence of invasive microorganisms. In contrast to B lymphocytes, T lymphocytes are unable to recognize bacteria or viruses in their native form. The structure of their antigen receptor only allows them to bind to small peptidic fragments that have to be stably presented by specific molecules at the surface of specific cells. These professional "antigen presenting cells" capture antigens and alert the immune system by expressing at their surface molecular complexes formed by their own major histocompatibility molecules (MHC) and fragments of the infectious agent. Extracellular microorganisms are captured by phagocytosis and digested into small peptides in the endosomal compartment of antigen presenting cells. The peptides able to bind to MHC class II molecules are transported to the cell surface. These antigen-MHC complexes are recognized by antigen specific CD4+ T lymphocytes, thus leading to the enhancement of antibody formation and of inflammatory responses which eliminate extracellular bacterial. In contrast, viruses or bacteria able to survive within the cytoplasm of the antigen presenting cells are digested by a specific multicatalytic enzymatic complex (the proteasome). The antigenic peptides released into the cytosol will be transported into the endoplasmic reticulum by an active peptide pump. The peptides able to bind to the groove of MHC class I molecules are transported to the cell surface. Their recognition by specific cytotoxic CD8+ lymphocytes leads to the destruction of the cells identified as infected. Thus, the mechanisms of antigen processing and presentation are able to generate a wide variety of antigenic fragments. Depending on the initial extra- or intracellular localization of the microorganism, some antigenic peptides will appear on the surface of antigen presenting cells on either MHC class I or class II molecules which are specifically recognized by either CD4+ or CD8+ lymphocytes. Only the antigenic peptides that are generated by this process and able to bind to MHC molecules of antigen presenting cells will be recognized by circulating lymphocytes and thus induce antigen specific immune response. Their identification therefore forms the basis of the defense mechanisms against infectious diseases and of novel immunization strategies. PMID- 8720321 TI - [Determination of systolic left ventricular time interval using the doppler technique. Analysis of aortic flow spectrum with transthoracic echocardiography]. AB - In 10 healthy volunteers (age 21-28 years; mean age 24.8 years +/- 1.9 years) systolic left ventricular time intervals (STI) were determined by analysis of aortic Doppler flow pattern. This method was compared with conventional calculation of STI, using electrocardiogram, carotid pulse curve and phonocardiography, which were registered in synchronicity to the Doppler examination. Both, CW- and PW Doppler echocardiography constantly showed slightly lower values than the conventional method. This referred to basic measurements as well as to measurements after the application of isoprenaline and quinidine. The PW Doppler method underestimated the conventional method concerning the electromechanic systole (QS2) by on average 3.98%, the left ventricular ejection period (LVET) by on average 2.24% and the pre-ejection period (PEP) by on average 7.3%. Using CW-Doppler-method, QS2 was on average 5.31% and LVET was on average 6.67% smaller than the values determined by conventional method, whereas PEP was overestimated by on average 1.69%. The study documented that positive and negative inotropic pharmacological effects were measured reliably by the Doppler echocardiographic method. Isoprenaline caused a significant shortening of frequency corrected QS2 (QS2c) from -55 +/- 17 to -85 +/- 20 ms (p < 0.05) using the PW-Doppler method; frequency corrected PEP (PEPc) was shortened from -40 +/- 14 to -67 +/- 14 ms (p < 0.05). The CW Doppler method also showed a statistically significant reduction of QS2c (from -64 +/- 18 to -89 +/- 25 ms; p < 0.05) and PEPc (from -37 +/- 16 to -64 +/- 12; p < 0.05). Likewise, the conventional method demonstrated statistically significant shortening of QS2c and PEPc after application of isoprenaline. LVETc did not change in a state of positive inotropy, no matter which method was used for determination. The negative inotropic effect of quinidine, measured by PW-Doppler, resulted in a prolongation of QS2c from -55 +/- 16 to -32 +/- 24 ms (p < 0.05) and of LVETc from -20 +/- 11 to +6 +/- 17 ms (p < 0.05). Using CW Doppler method, quinidine led to a lengthening of QS2c from -63 +/- 16 to -42 +/- 22 (p < 0.05) and of LVETc from 33 +/- 11 to -12 +/- 8 ms (p < 0.05). The conventional method also demonstrated a statistically significant increase of QS2c and LVETc. None of the 3 methods in question showed a statistically significant alteration of PEPc in the negative inotropic state. CONCLUSIONS: Doppler-echocardiographic analysis of aortic flow pattern constitutes a new method for the measuring of systolic time intervals. Basic values as well as changes due to positive or negative inotropic effects are reliably determined, in comparison with conventional methods the measurements are slightly lower. PMID- 8720323 TI - [Antibiotic resistance in pneumococci]. AB - In 1875, 7 years prior to the description of the Koch bacillus, Klebs visualized the first Streptococcus pneumoniae in pleural fluid. Since then, this organism has played a decisive role in biomedical science. From a biological point of view, it was extensively involved in the development of passive and active immunization by serotherapy and vaccination respectively. Genetic transformation was also first observed in S. pneumoniae, leading to the discovery of DNA. From a clinical point of view, S. pneumoniae is today still a prime cause of otitis media in children and of pneumonia in all age groups, as well as a predominant cause of meningitis and bacteremia. In adults, bacteremia still has a mortality of over 25%. Although S. pneumoniae remained very sensitive to penicillin for many years, penicillin-resistant strains have emerged and increased dramatically over the last 15 years. During this period the frequency of penicillin-resistant isolates has increased from < or = 1% to frequencies varying from 20 to 60% in geographic areas as diverse as South Africa, Spain, France, Hungary, Iceland, Alaska, and numerous regions of the United States and South America. In Switzerland, the current frequency of penicillin-resistant pneumococci ranges between 5 and > or = 10%. The increase in penicillin-resistant pneumococci correlates with the intensive use of beta-lactam antibiotics. The mechanism of resistance is not due to bacterial production of penicillinase but to an alteration of the bacterial target of penicillin, the so-called penicillin binding proteins. Resistance is subdivided into (1) intermediate level resistance (minimal inhibitory concentration [MIC] of penicillin of 0.1-1 mg/l) and (2) high level resistance (MCI > or = 2 mg/l). The clinical significance of intermediate resistance remains poorly defined. On the other hand, highly resistant strains have been responsible for numerous therapeutic failures, especially in cases of meningitis. Antibiotics recommended against penicillin-resistant pneumococci include cefotaxime, ceftriaxone, imipenem and in some instances vancomycin. However, penicillin-resistant pneumococci tend to present cross-resistances to all the antibiotics of the beta-lactam family and could even become resistant to the last resort drugs mentioned above. Thus, the explosion of resistance to penicillin in pneumococci is a ubiquitous phenomenon which must be fought against by (1) avoiding excessive use of antibiotics, (2) the practice of microbiological sampling of infected foci before treatment, (3) the systematic surveillance of resistance profiles of pneumococci against antibiotics and (4) adequate vaccination of populations at risk. PMID- 8720324 TI - [Prenatal and perinatal infections--problems for the practicing pediatrician: group B streptococci, varicella, toxoplasmosis]. AB - A practical approach is reported for the care of the neonate born to a mother infected/colonized during pregnancy by group B streptococcus, varicella-zoster virus or Toxoplasma gondii. Starting from clinical situations, an attempt is made to work out evidence based recommendations using an overview of the current literature. GROUP B STREPTOCOCCI: Relevant factors for the treatment of infants born to colonized mothers are clinical symptoms, gestational age, additional risk factors (such as premature rupture of membranes or maternal fever) and intrapartum antibiotics. Postnatal antibiotic prophylaxis and laboratory screens failed the test of controlled trials. Transfer to a neonatology unit is recommended for symptomatic term and all preterm infants. Asymptomatic term infants should be carefully monitored during the first 48 hours for signs of respiratory, circulatory or thermoregulatory compromise. VARICELLA: In the case of maternal varicella near term, delaying delivery for one week will lower the risk of severe neonatal varicella. The postnatal administration of varicella zoster-immunoglobulin to the neonate is supported by some (if limited) evidence from the literature in the case of maternal eruption between 7 days before and 2 days after delivery. In newborns of mothers with eruption appearing later immunoglobulin is often recommended, though no supporting clinical evidence is available. There are no data to justify the use of immunoglobulin after exposure during pregnancy in order to prevent pneumonia in the pregnant patient, but there are preliminary indications that its application could lower the risk of congenital varicella syndrome (2% between 13 and 20 weeks). The use of immunoglobulin in very low birth weight infants after nosocomial exposure is generally recommended but efficacy data are lacking. TOXOPLASMOSIS: The practical approach depends on clinical findings in the newborn and laboratory results during pregnancy and after birth. Examination of the newborn should include fundoscopy, cranial sonography and, in cases of documented infection, lumbar puncture. Serology from cord blood comprises assays for IgG, IgM and if possible IgA/IgE. If available, demonstration of the parasite by culture or PCR can be helpful. All infants with documented congenital toxoplasmosis should be treated for a minimum of 12 months. In the case of suspected toxoplasmosis the child should be treated as long as the suspicion persists. The prognosis after consequential therapy is less bleak than previously reported for untreated children even in seriously symptomatic patients. PMID- 8720325 TI - [Muscle sympathetic nerve activity in patients with spinocerebellar degeneration without orthostatic hypotension]. AB - Muscle sympathetic nerve activity (MSNA) was compared in 6 patients (age; 45-76 years old, 3 males and 3 females) with spinocerebellar degeneration (SCD) without orthostatic hypotension (OH) and in 10 (age; 38-78 years old, 3 males and 7 females) with other neurologic disorders (controls). MSNA was elicited from the peroneal nerve at the resting recumbent position by using a microneurographic method. MSNA, heart rate (HR) from the electrocardiogram and mean blood pressure (MBP) measured by Finapres were recorded simultaneously at rest and during the head-up tilting at 45 degrees. Although there were no significant differences in MBP and HR between both groups, burst rate (bursts per minute) and burst incidence (bursts per 100 heart beats) of MSNA in SCD patients were significantly decreased as compared with controls (p < 0.01). During the head-up tilting, responses in MBP and MSNA showed a tendency to decrease in SCD patients. We concluded that the sympathetic noradrenergic system might be potentially impaired in SCD patients without OH. PMID- 8720326 TI - [Three cases of clinically diagnosed corticobasal degeneration--neuroimaging studies with MRI, SPECT and PET]. AB - We report three cases of clinically diagnosed corticobasal degeneration: a 63 year-old male, a 63-year-old female and a 62-year-old female. All the three patients exhibited asymmetric limb-kinetic apraxia and extrapyramidal signs. Neuroimaging studies with MRI, 123I-IMP SPECT and FDG-PET were performed in each of the three cases and FDOPA-PET was performed in patient 1. MRI analysis showed asymmetric cortical atrophy, especially in the parietal cortex, in all the three cases. 123I-IMP SPECT analysis revealed asymmetric reduction of the cerebral blood flow, in accordance with the MRI findings. FDG-PET analysis showed marked asymmetric reduction (> or = 10%) of cerebral metabolic rates for glucose (CMRGlc) in the lateral frontal cortex, lateral posterior frontal cortex and primary motor and sensory cortex in all the patients, although the asymmetry in the thalamus which had been described as a characteristic CBD finding in previous reports was not observed in any of the patients in our study. FDOPA-PET analysis revealed reduction of uptake in both the putamen and caudate nucleus, predominantly in the caudate nucleus, which suggests that the FDOPA-PET findings correlate with the extrapyramidal signs observed in the case of the 63-year-old male. PMID- 8720327 TI - [Cardiac dysfunction in female gene carriers of Duchenne muscular dystrophy]. AB - Cardiac dysfunction and its correlation with skeletal muscle dysfunction were examined in 16 definite female gene carriers of Duchenne muscular dystrophy (DMD). Five out of 16 carriers (31.3%) had cardiac symptoms and 8 carriers (50.0%) showed an increased cardio-thoracic ratio on chest X-ray. Electrocardiographic abnormalities including a high R:S ratio (> or = 1.0) in the V1 lead, deep Q wave (> 3 mm) in the I, II, aVL, V5, and V6 leads, complete right bundle branch block and premature ventricular beats, were observed in 9 carriers (56.3%). On echocardiographic examination, an increase in the end-diastolic dimension of the left ventricle and a decrease in the ejection fraction suggestive of dilated cardiomyopathy were found in 12 carriers (75.0%). Tl-201 myocardial SPECT scan was performed in 2 symptomatic carriers and showed an area of hypoperfusion in the inferio-posterior wall. These findings were similar to previously reported findings in DMD patients. A biopsy of the myocardium was obtained in one carrier with her informed consent for the biopsy. Immunohistochemical staining demonstrated that 75.4% of the myocardial fibers were negative for dystrophin, suggesting that her cardiac dysfunction is caused by the abnormal expression of dystrophin in the cardiac muscle. On examination of the skeletal muscle function, none of the carriers had clinical evidence of muscle weakness or atrophy. However serum creatine kinase activity was elevated in 14 of 16 carriers (87.5%). Computed tomography (CT) of the lower limb muscles demonstrated widened spaces among muscles and moss-eaten appearance of low density areas within muscles and CT value was decreased, suggesting the subclinical involvement of the skeletal muscle. In the carriers without cardiac symptoms, there was a negative correlation (p < 0.05) between the end-diastolic dimension of the left ventricle and the CT value of the biceps femoris muscle (a muscle with the lowest CT value among the lower limb muscles). This indicates that there was an apparent correlation between the cardiac and skeletal muscle dysfunction. These findings suggest a high frequency of clinical and subclinical involvement of the cardiac and skeletal muscles in DMD carriers. To protect them from cardiac failure, cardiac dysfunction in DMD carriers needs to be examined closely and treated appropriately before the carriers become symptomatic. PMID- 8720328 TI - [Regional cerebral glucose metabolism associated with ataxic gait--an FDG-PET activation study in patients with olivopontocerebellar atrophy]. AB - We studied changes in regional cerebral glucose metabolism during bipedal gait in 7 patients with olivopontocerebellar atrophy (OPCA) and 7 control subjects without neurological disease. All OPCA patients in this study were able to walk on the treadmill without a support or handrails. We used 18F-2-fluoro-2-deoxy-D glucose (18F-FDG) and positron emission tomography (PET) to evaluate glucose metabolism under two different conditions; 30 minutes' treadmill walking, and supine resting. The two sets of PET images were three-dimensionally registered to the magnetic resonance image (MRI) in each subject. Then, the PET images were normalized by the global value. Regions of interest (ROIs) were drawn on the cerebellar vermis, cerebellar hemispheres, pons, and thalamus, and FDG uptake was obtained to calculate the activation ratio (= [FDG uptake under walking]/[FDG uptake under resting]) for each region. We used unpaired t-test for statistical analysis to compare the OPCA patients and the normal control subjects. Normalized resting FDG uptake had no significant difference between control subjects and OPCA patients in any region. Activation ratio of OPCA patients was significantly decreased in the cerebellar vermis compared with the control subjects. In the control subjects, FDG uptake had little difference between resting and walking in the cerebellar hemisphere, pons and thalamus. On the other hand, the FDG uptake of OPCA patients was moderately increased by walking in these regions. In the cerebellar vermis, in spite of no significant difference between the normals and the patients during resting, the activation ratio in OPCA patients was significantly decreased. We speculate that the reduction of activation ratio in the cerebellar vermis reflects the dysfunction caused by degeneration. The result suggests that the PET activation study can demonstrate cerebellar dysfunction in the early phase of OPCA, in which other neuro-imaging methods cannot detect the tissue atrophy, hypometabolism or hypoperfusion in the resting state. In the cerebellar hemisphere, pons and thalamus, the activation ratio was nearly equal to one in control subjects, while it was larger in OPCA patients. We speculate that the instability during the ataxic gait increases the inputs from the vestibular, somatosensory and visual systems to these regions and outputs from these regions to the other neural systems. In conclusion, PET activation study is a useful and noninvasive technique for investigating the brain function associated with human gait. PMID- 8720329 TI - [Neuropsychological comparison between dementia of Alzheimer type and multiple subcortical infarction by easily applicable test battery]. AB - We compared the neuropsychological findings between dementia of Alzheimer type (DAT) and multiple subcortical infarction (MSI) using easily applicable test battery. Patients with DAT (N = 27) fulfilled the criteria of NINCDS-ADRDA for probable DAT and belonged to the mild degree in the DSM-III-R criteria for dementia. Patients with MSI (N = 30) scored more than 7 points in Hachinski Ischemic Score and were physically independent (Rankin Disability Scale < or = 3). Patients with DAT showed significantly more impaired performance than MSI in Mini-Mental State Examination, copy and delayed recall of Rey-Osterrieth Complex Figure Test and trial 5 of Rey Auditory-Verbal Learning Test, while patients with MSI achieved fewer categories and made more perseverative errors on Wisconsin Card Sorting Test than patients with DAT. Among these tests, delayed recall of Rey-Osterrieth Complex Figure Test seemed to be the most sensitive to discriminate DAT from MSI. These findings were thought to be compatible with the pathological facts that the degenerative process progresses from temporo-parietal lobe in DAT and frontal-subcortical region in MSI. PMID- 8720330 TI - [Muscle afferent block for the treatment of writer's cramp]. AB - A 29-year-old man suffered from dystonic writer's cramp for over three years. When he wrote, typed and did other tasks using right hand, dystonic involuntary movement triggered medial rotation of the arm, wrist extension and shoulder elevation. Medication, biofeedback, and botulinum injection were performed without much success. We tried to block the sensory input from muscles by using lidocaine and ethanol. We made injections of 0.5% lidocaine 50ml and 99% ethanol 5ml into muscles with abnormal activity at the frequency of twice a week for about six months. After the treatment, dystonic movement was remarkably improved and he was then able to write, type and perform other tasks with the right hand. Side effects included pain of the injection site, nausea and dizziness, which lasted for a few hours. This "muscle afferent block" did not cause muscle weakness. We speculate that muscle afferent plays a pivotal role in dystonia so that its blocking may be of clinical use. PMID- 8720331 TI - [Serial changes of MRI and SPECT findings in a case of adult-onset SSPE]. AB - A 20-year-old man who had developed involuntary movement of his left hand and memorial disturbance visited our hospital in December, 1991. On admission, myoclonus, dementia and speech disturbance were recognized. He was diagnosed as subacute sclerosing panencephalitis (SSPE) based on a high titer of serum anti measles antibody (1/256), serum anti-measles-IgG antibody (> 1/4,800) and typical EEG fiding of periodic synchronus discharge (PSD). Inosine pranobex was administrated orally (4,800mg per day). Serial cranial magnetic resonance imagings (MRI) were taken since January, 1992 to June, 1994. No abnormal finding was demonstrated until April 16, 1992 in MRI, but 123I-IMP SPECT detected decreased accumulation in parietal to occipital lobes on early image in February 5, 1992. Marked high signal area on T2 weighted image in right temporal lobe and parieto-occipital lobe were firstly demonstrated in June 22, 1992 on MRI. These high signal lesions alternated the areas and locations, but the changes were not related to his clinical symptom. These findings may suggest ischemic changes after demyelination. His symptoms have been improving gradually since June, 1994. To our knowledge, 42 cases of adult-onset SSPE were reported so far (5 were in Japan). This case is the first report in the world on adult-onset SSPE serially observed with MRI and SPECT since early stage. PMID- 8720332 TI - [A case report of variegate porphyria]. AB - We reported a 30-year-old woman with variegate porphyria. Family history disclosed that her mother had the same skin lesions as the present case. She has had repeated vesicles and brownish pigmentation on her upper extremities since eighteen years of age. Recently she was suffering from common cold, and was treated with drugs. One week later, she started to develop weakness and dysesthesia in all limbs, hallucination, and urinary disturbance. Three weeks later, she was admitted to our hospital because she could not stand any more. On general examination, she had brownish pigmentations and crusts in hands and forearms. Neurologically she had moderate weakness, dysesthesia and hyperreflexia in all limbs. Urine was dark-red. Head and cervical MRI was normal. Nerve conduction studies and somatosensory evoked potentials were within normal limits. Excretion of porphyrins, especially protoporphyrin and X-porphyrin in the feces were markedly increased, and the diagnosis of variegate porphyria was made. Intravenous administration of glucose was effective and her symptoms began to improve in two weeks after her admission. However, recovery of weakness took two months and dysesthesia did not disappear in three months. We considered that her clinical manifestations were caused by involvement of the central and peripheral nervous systems. PMID- 8720333 TI - [A case of recanalized basilar artery following basilar artery occlusion due to spontaneous dissection]. AB - An 85-year-old man who did not have any previous history of heart disease was admitted to our hospital with the chief complaints of disturbance of consciousness, tetraparesis, and bilateral Babinski signs. On admission, intravenous digital subtraction angiography (DSA) showed bilatral vertebral arteries without filling the basilar artery. On the second hospital day, MRI showed a septum in the basilar artery, and no infarctions were noted in the brain stem. He gradually improved with heparin therapy. On the tenth hospital day, intravenous DSA disclosed filling of the basilar artery. On the sixteenth hospital day, conventional cerebral angiography revealed filling of the basilar artery from the left vertebral artery, and string sign and linear shadow (intimal flap) was noted in the basilar artery. He was diagnosed to have the basilar artery dissection. The patient became ambulatory and was discharged independently. This is very rare case of the basilar artery dissection with a good prognosis. PMID- 8720334 TI - [A case of hemichorea presenting with inattention and denial of involuntary movement]. AB - We report a case of hemichorea presenting inattention and denial of involuntary movement. A 64-year-old right-handed man was hospitalized with acute hemichorea on the right side of the body. The patient had hemichorea which was frequently followed by purposeful movement such as fumbling with his glasses or combing his hair. He claimed that the involuntary movements were habits showing both inattention and denial of the involuntary movements. These symptoms were very unique. On MRI of the brain, T2 elongated spots were detected at the bilateral pallidum, suggesting occulusive cerebrovascular disease. On 123I-IMP-SPECT hypoperfusion was observed at the right cerebral basal ganglia extending to the right medial frontal lobe. We supposed that this inattention and denial of involuntary movements were caused by dysfunction of the system connecting the medial frontal lobe with basal ganglia which was presented as hypoperfusion on SPECT. PMID- 8720335 TI - [Meningeal seeding of spinal cord glioblastoma multiforme without any signs of myelopathy]. AB - An autopsy case of meningeal spreading of glioblastoma multiforme (GBM) probably originating in the cervical cord was reported. In contrast to autopsy findings, main symptoms were similar to subacute meningitis, and any signs of myelopathy could not be detected during the clinical course. The patient was a 22-year-old man who was hospitalized because of a 2-week history of progressive headache following cough and slight fever. Vomiting and somnolence, developing 5 days before admission, were improved the day after a lumbar puncture performed at another hospital. On admission, meningeal signs, mild right abducens palsy, and depressed deep tendon reflexes were detected. There was no muscle weakness, sensory loss, or Babinski sign. Lumbar puncture yielded CSF with an opening pressure of 280 mmH2O, 21 mononuclear cells/mm3, a protein level of 645 mg/dl, and a glucose level of 7 mg/dl. Cytology for malignancy and multiple cultures were negative. Brain CT scan showed mild hydrocephalus and swelling of the brainstem and cerebellum. Intravenous administration of antimicrobial drugs was started and ventriculoperitoneal shunt surgery was performed. During the third hospital week, however, meningeal signs progressed and somnolence reappeared, followed by progressive multiple cranial neuropathy and polyradiculopathy characterized by flaccid tetraparesis, muscle atrophy, and sensory impairment without a level. Babinski sign could not be detected. MRI revealed an intramedullary lesion in the lower cervical cord, swelling of the brainstem, cerebellum, spinal cord and nerve roots, and a diffuse or nodular thickning of leptomeninges. Repeated CSF cytology disclosed atypical cells. Examinations for extraneural malignancies were negative. During the 9th hospital week, flaccid tetraplegia progressed and stupor developed, and the patient died 2 weeks later. The pathological study was limited to the brain. The brain showed a diffuse opalescent thickening of the leptomeninges, especially over the ventral aspect of the brainstem and cerebellum, where the blood vesseles and cranial nerves were obscured. Histological examination revealed the appearance of GBM. The malignant cells filled the subarachnoid space, and to a variable extent penetrated the brainstem and cerebellum along perivascular spaces. Hypertrophied optic tracts and trigeminal nerves were also infiltrated by the cells. However, there were no mass lesions assumed to be primary ones anywhere in the cerebral parenchyma. Therefore, it was thought that GBM primarily growing in cervical cord metastasized to intracranial subarachnoid space by way of the cerebrospinal fluid pathway. Spinal cord GBM usually presents signs of myelopathy from the early stage. The present case was characterized by no signs of myelopathy during the clinical course. It is speculated that the intramedullary GBM, originating near the surface of cervical cord, had been rapidly disseminated into the subarachnoid space up to the intracranial cavity before myelopathy appeared, and caused cranial and spinal nerve roots dysfunction, which covered signs of myelopathy. Cord GBM should be always considered as a differential diagnesis in a case of subacute meningitis. PMID- 8720336 TI - [A case of severe sarin poisoning in the sarin attack in Matsumoto--one-year follow-up of clinical findings, and laboratory data]. AB - A patient severely poisoned with sarin in the Sarin Attack in Matsumoto is described. A 19-year-old man was exposed to sarin at 23:00 on 27 June, 1994. At 1:00 of the following day, a rescue team found and brought him to the hospital. His blood pressure was 150-80mmHg and the heart rate was 120/min with frequent premature ventricular contractions (PVC). His respiration was shallow and copious salivation and excretion from the respiratory tract were observed. Consciousness disturbance, generalized convulsion, severe miosis and fasciculation of tongue, facial muscle and extremities were also marked. Serum cholinesterase was 21 U/l (normal 109-249) and acetylcholinesterase in erythrocyte (E-AchE) was 0.1U/l (normal 1.2-2.0). Electroencephalogram (EEG) 30 hours after exposure showed polispike and wave complexes. Ventilatory assistance, forced urination and injection of diazepam and atropin improved his general condition and he was discharged 18 days after exposure. Three months after exposure, E-AchE was normalized and there was no complaint. But one year after exposure, EEG showed epilecpic discharges during sleep, and Holter electrocardiogram showed frequent PVC. As no clinical cases of severe sarin poisoning like this patient was reported, a longterm follow-up of this patient is very important. PMID- 8720337 TI - [A patient with Kennedy-Alter-Sung syndrome showing cardiomyopathy]. AB - A 31-year-old man with a history of progressive muscular atrophy and weakness from around 22 years of age, recently experienced arrhythmia. On examination he showed gynecomastia and slight proximal weakness in both upper and lower extremities. Androgen receptor gene analysis showed an increased number of tandem CAG repeats in exon 1, thus leading to a diagnosis of Kennedy-Alter-Sung (KAS) syndrome. Cardiological investigations including echocardiography, scintigraphy and catheterization disclosed dilated cardiomyopathy. Cardiac muscle biopsy showed myocardial cell degeneration. KAS syndrome is causatively related to androgen receptor gene abnormality. This receptor is widely distributed throughout human body, including the genital tract, spinal cord, liver, and heart. Androgen receptor dysfunction may induce cardiac muscle involvement in patients with KAS syndrome, producing the previously unknown pathophysiology of cardiomyopathy. PMID- 8720338 TI - [Lower level of endothelin-1 in migraine with aura]. AB - Endothelin-1 (ET-1) exerts powerful vasoconstrictive and blood pressure elevating properties through endothelial cells. However, no systematic examination of ET-1 in migraine has ever been attempted. The present investigation was focused on evaluating the level of ET-1 in patients with migraine with aura. Studies on ET-1 were made in 17 patients with migraine with aura (age, 23.4 +/- 9.1 years old, mean +/- SD) according to the Classification of Headache of the Ad Hoc Committee. All patients had been free of migraine attacks for at least 7 days prior to the examination. Twenty-eight age-matched healthy volunteers (age, 23.0 +/- 12.3 years old) were similarly used as a control group. None of them revealed evidence of any other disease, such as hypertension, obesity, or heart disease. Informed consent was obtained from each subject. We measured the immunoreactive ET-1 in plasma by radioimmunoassay. The plasma level of ET-1 in migraine was 2.53 +/- 1.06 pg/ml. On the other hand, the level of ET-1 in the controls was 4.24 +/- 0.80 pg/ml. The ET-1 level in migraine was significantly lower than that in the controls (p < 0.002). We also measured the level of ET-1 in the cerebrospinal fluid (CSF). There was no significant difference between migraine (23.2 +/- 3.10 pg/ml) and the control group (20.85 +/- 3.20 pg/ml). In conclusion, the lower plasma level of ET-1 observed in the patients with migraine is consistent with the pathogenesis of migraine, further supporting the hypothesis that a lower ET-1 may be closely related to marked vasodilatation following constriction partly due to a deficiency of ET-1 for maintaining vasoconstriction. PMID- 8720339 TI - [Chorea with prominent spasticity associated with an expansion of the CAG trinucleotide repeat in the IT15 gene: a case report]. AB - A 60-year-old female, who had been noted to have slowly progressive mental changes since the age of 40, subsequently developed irregular, abrupt, jerking movements of her limbs, and coordination and walking difficulty after 15 years. Her father, aunt and sister had also been suffering from similar involuntary movements and changes of personality. Neurological examination revealed slight dementia, choreic movements and the evidence of pyramidal tract signs; i.e., she showed general hyperreflexia of extremities, bilateral ankle clonus, and extensor plantar responses of lower extremities. laboratory data were all within normal ranges except iron deficiency anemia. Axial MRI showed moderate cortical atrophy, however, without dilatation of the frontal hours of lateral ventricles and the caudate head appeared normal. Our initial impression was dentatorubropallidoluysian atrophy (DRPLA). However, the genetic analysis of her peripheral leukocytes disclosed an expansion of the CAG repeats (43 copies) in the IT15 gene in chromosome 4p, indicating the diagnosis of Huntington's disease (HD). CAG repeats in chromosome 12, known to link with DRPLA, was not expanded. Thus, this case is unique in that the classical symptomatologies of HD was betrayed by the presence of prominent pyramidal tract signs. This patient may be of importance in demonstrating the diversity of neurological symptoms and neuropathologies caused by CAG trinucleotide repeats in HD. PMID- 8720340 TI - [A case of cerebral infarction in a young adult following an abuse of cocaine and other drugs]. AB - We reported a case of cerebral infarction in a young adult following an abuse of cocaine and other drugs. A 27-year-old male was admitted to our hospital because of the abrupt onset of right hemiplegia and dysarthria. Brain CT and MRI examinations showed cerebral infarction in the region of the left caudate head and the left corona radiata. Cerebral angiography revealed total occlusion at the origin of the left middle cerebral artery, with neither vascular malformation nor atherosclerotic change. There were no obvious causes that might lead to cerebral infarction in young adults, including potential cardiac sources of embolism, abnormalities of blood coagulation, or collagen vascular diseases. From his history we learned he had started abusing cocaine and other drugs at the age of 16 years, and that he had been addicted to these drugs up to at least 2 months earlier than the onset of cerebral infarction, so we suggested his drug abuse might have affected the occurrence of cerebral infarction. We would like to emphasize that using 'drugs' should be taken into consideration as a possible cause of cerebral infarction in young adults. PMID- 8720341 TI - [Identification of anti-neuronal antibody]. PMID- 8720342 TI - [Delayed diagnosis: inevitable fatality or wrong methodological approach? Churg Strauss syndrome]. AB - A case of CSS is reviewed, along its difficult diagnosis, to check if its late discovery is attributed to inevitability or to erroneous approach. PMID- 8720343 TI - [RS3PE syndrome. A new rheumatic inflammatory syndrome in advanced age]. AB - RS3PE syndrome (Remitting Seronegative Symmetrical Synovitis with Pitting Edema) is a new form of arthritis in the elderly, characterized by symmetrical polysynovitis of acute onset associated with distal pitting edema. Tests for serum IgM rheumatoid factor are typically negative. RS3PE syndrome affects Caucasians aged over 60, predominantly males. It has a benign course and remits completely without residua. PMID- 8720344 TI - [Reflexions on the Tietze syndrome. Clinical contribution]. AB - The authors report 4 cases of Tietze's syndrome. In 3 of them the syndrome onset during a rheumatic disease (psoriatic arthritis, sternoclavicular hyperostosis). It is thus necessary to distinguish between the idiopathic of unknown ethiology and the secondary form due to, a well defined disease, in 3 cases observed. PMID- 8720345 TI - [Morphometric analysis of mammographic images in clinical oncology]. AB - The three standard projections relative to mammographies of 40 patients with breast cancer pre-operatively classified as T1-T2 were analyzed by computerized morphometric elaborations. This method si able to better define view of images, permetting pre-operative connection of staging, with an increment of 30% if confronted with histologic TNM (pTNM). The utilization of method is discussed to evaluate the best way for the local definition of tumor extension, permitting to use a correctly conservative surgery (quadrantectomy, tumorectomy) in a greater number of patients. PMID- 8720346 TI - [Radiologic evaluation of parietal infiltration of bladder cancer (integrated imaging: US, TC, RM) and comparison with transurethral resection (TUR)]. AB - In this work we checked the reliability of Ultrasounds (US), Computed Tomography (CT) and Magnetic Resonance Imaging (MRI) in the staging of parietal infiltration of bladder carcinoma. We studied 49 patients with documented bladder carcinoma. All patients underwent CT examination, 43 patients to MR examination and 40 patients to US examination. Between the patients who underwent US examination, 35 out of them had a sovra-pubic Ecography, 27 patients a trans-rectal Ecography and in 30 patients a trans-uretral Ecography was performed. After radiologic examination, all patients underwent a deep fractionated trans-uretral resection of the neoplastic mass (TUR) for the tumor staging. The results of the computed images and US examinations were then compared with the TUR staging. Among the group of patients studied with US, the agreement between imaging and TUR staging was respectively of 51% for the sovra-pubic study, 44.5% for the trans-rectal study and 56.7% for the trans-uretral study. The agreement was of 55% with the CT studies and 45.7% with the MRI. The sovra-pubic US studies showed a good correlation in the evaluation of superficial lesions (Tis-T1 stadied on TUR), if compared to the CT and MRI studies; moreover US is certainly more suitable because of its lower price and discomfort for the patient. On the contrary patients with a documented stage > T1 need a CT and MRI study for the correct staging. MRI has shown to be more sensitive in selected cases with localization of tumor on the top of the bladder roof, on its floor or in the follow-up after TUR. CT scan showed a better reliability in the staging of superficial lesions (Tis-T1 stages). On the contrary MRI has shown to be more sensitive in distinguishing superficial lesions (T1-T2 stages) from deeper lesions with muscles infiltration (T3a stage) on the basis of the loss of the parietal hypointense line on the T2 weighted images. Tumors with a grade T3b or more are well studied in both CT scan and MRI, which allow to get more useful information than US about the lynphoglandular involvement. PMID- 8720347 TI - [Biocompatibility and extracorporeal circulation]. AB - Present knowledge about biocompatibility during extracorporeal circulation, especially in hemodialysis, has been reported. Clinical implications of membrane biocompatibility on coagulation, fibrinolysis, complement and kalikrein cascade has been analyzed. Particular attention is for the platelet activation in hemodialysis and ist effect upon the coagulation system and dialytic efficiency. The the role of epoprostenol sodium salt (prostacyclin) continous infusion in hemodialysis has been described. Finally future perspectives of anticoagulation during dialytic treatment have been advanced. PMID- 8720348 TI - [Evaluation of the efficacy of oral sumatriptan in the management of migraine attacks. Clinical results]. AB - This italian multicentre, double blind, parallel groups study compared the efficacy, safety and tolerability of oral sumatriptan, given as new film-coated tablet, with placebo in the acute treatment of migraine. 88 Patients received placebo and 162 patients received sumatriptan 100 mg (plus an optional dose 2 h later if the headache persisted plus a further optional dose for recurrence within 24 h). Sumatriptan was significantly more effective than placebo at releiving headache at 2 h (51% versus 31%, P = 0.003) and 4 h (71% versus 35%, P < 0.001). Fewer sumatriptan-treated patients required a second dose compared with placebo-treated patients (49% versus 74%, P < 0.001). Sumatriptan was more effective than placebo at relieving nausea, vomiting and photophobia/phonophobia. Few patients were evaluable for treatment of headache recurrence, and statistical analysis was not possible. More sumatriptan-treated patients than placebo-treated patients reported adverse events (29% versus 16%) but the difference was not statistically significant. More of these events were mild to moderate in severity, of short duration and resolved without treatment. Sumatriptan had no clinically significant effect on blood pressure, heart rate, electrocardiogram or laboratory test results. It is concluded that oral sumatriptan 100 mg, given as a film-coated, tablet, provides an effective and well-tolerated acute treatment for migraine. PMID- 8720349 TI - [Metabolic tolerance of isradipine. Evaluation after three months of therapy]. AB - The aim of the present study was to evaluate efficacy on blood pressure values and metabolic tolerability of Isradipine, which was given to 15 mild hypertensive non diabetic subjects (average age 63.8 +/- 10.9 years), at the dosage of 5 mg once a day. Plasma lipids concentrations and oral glucose tolerance test (OGGT) with plasma insulin assay were carrid out before, after one and three months of therapy. The statistical analysis was done using the Student's t test for paired data. Isradipine showed a good efficacy on lowering high blood pressure. In no patients the drug induced impaired glucose tolerance. Isradipine had contrasting, but not statistically significant effects on lipid concentrations, namely, decrease of triglycerides, increased of any form of cholesterol (total, HDL, LDL), apoproteins and lipoprotein(a). PMID- 8720350 TI - [Current orientation on medical therapy of Crohn disease]. AB - The Crohn's disease has an increasing incidence and prevalence in the western countries and also in Italy; genetic, familiar and enviromental factors, but, above all, altered immunity have pathogenetic effects for the chronic flogosis and damage of gastroenteric tract. With ulcerative rettocolitis, the Crohn's disease is part of the chronic inflammatory bowel disease. In the present paper the medical therapy of Crohn's disease is especially studied; it is based on the use of corticosteroids and SASP or its active metabolite 5-ASA. Other pharmacs are reviewed as: immunosuppressors, antibiotics, antimetabolites and symptomatic or substitutive therapy. At the end the authors pointed out therapeutic lines in relation to acute or chronic stage and gravity of the disease. PMID- 8720351 TI - [High resolution CT in the evaluation of pulmonary parenchymal diseases]. AB - In this manuscript we evaluate the role of HR CT in polmonary parenchymal disease. PMID- 8720352 TI - [The quality of life after extirpation of the rectum for carcinoma]. AB - BASIC PROBLEM AND AIM OF STUDY: The quality of life for patients who have undergone total rectal resection for carcinoma is impaired by the artificial intestinal stoma. Their psychosocial disorders were analysed in relation to the method of looking after the stoma (with or without controlled stool emptying by intestinal irrigation) and compared with patients who had a after continence preserving operation for rectal carcinoma. PATIENTS AND METHODS: Of the 205 patients (aged 58.6 +/- 8.1 years; 72 women and 133 men) 78 had a continence resection (group 1), 127 had a colostomy (irrigation in 81, group 2a; merely looking after the colostomy bag in 46, group 2b). Their personality characteristics were tested prospectively with the State-Trait Anxiety Inventory (STAI) and quality of life parameters with a standardized questionnaire. RESULTS: While personality traits were similar, significantly fewer patients in group 2a complained about disturbances of self esteem (32.9 vs 60.0%; P < 0.01), and had pessimistic future expectations (20.5 vs 46.5; P < 0.01) than those in group 2b. There was no significant difference in these two parameters between groups 1 and 2a. The irrigation procedure was taught to 60.5% of patients during their hospital stay. Accepting irrigation was more common when rehabilitation was begun early rather than delayed (72.9 vs 42.9%; P < 0.01). Disturbances of erection occurred in 69.9% of men younger than 60 years after the operation. CONCLUSIONS: Compared with conventional bag care, regular irrigation improves the quality of life for patients with a colostomy. Postoperatively disordered erection is an independent risk factor for any abnormal self esteem and depression. They should be stressed more during history taking and therapeutically. PMID- 8720353 TI - [Coarctation of the abdominal aorta]. AB - HISTORY AND FINDINGS: When a 27-year-old woman went to her general practitioner with symptoms of acute gastroenteritis he noted a paraumbilical murmur on auscultation. There were no symptoms of intermittent claudication or abdominal angina. All peripheral pulses were easily palpable and bilaterally equal, and there were no vascular murmurs. Physical examination and blood pressure (140/70 mm Hg bilaterally) were normal. INVESTIGATIONS: Biochemical tests gave no indication of inflammatory disease. Oscillography showed a brief decrease in amplitude after muscular exertion, predominantly of the thigh. Posterior tibial systolic pressure was 20 mm Hg lower than radial pressure on the right, 25 mm Hg on the left. Colour Doppler sonography demonstrated elongated and looping coarctation of the abdominal aorta. TREATMENT AND COURSE: As the patient had no symptoms only regular follow-up was indicated. There have been no symptoms for 27 months and no progression of the coarctation. CONCLUSION: In young patients with the described symptoms abdominal coarctation should be included in the differential diagnosis. PMID- 8720354 TI - [Acute kidney failure in a recurrence of sarcoidosis at the height of summer]. AB - HISTORY AND FINDINGS: A 35-year-old woman with sarcoidosis, successfully treated 3 years previously, developed fatigue, nausea and loss of weight during a summer holiday in Italy where she was exposed to much bright sun. There had been no previous recurrence of the sarcoidosis. On return she was found to be in renal failure with marked hypercalcaemia. Physical examination merely revealed peasized non-tender submandibular and cervical lymph nodes, but was otherwise unremarkable. INVESTIGATIONS: Serum creatinine and calcium concentrations (3.8 mg/dl and 3.6 mmol/l) were markedly elevated. Computed tomography showed an enlarged submandibular lymph node, but no other organ abnormality. Biopsy of the salivary gland demonstrated recurrent sarcoidosis. TREATMENT AND COURSE: Calcium concentration became normal (2.4 mmol/l) within two weeks of treatment with methylprednisolone (100 mg daily for 3 days, followed by gradual reduction to 30 mg daily for 3 months, until discontinued slowly). Creatinine concentration, which had fallen to 1.5 mg/dl, was at the upper limit of normal (1.1 mg/dl) after 4 months and 0.8 mg/dl after 8 months. CONCLUSION: Marked exposure to UV light should be avoided in patients with a history of recent sarcoidosis, because it increases the formation of vitamin D3 precursors: In the case of recurrent sarcoidosis, these precursors are converted to 1,25 dihydroxyvitamin D3 in the sarcoidosis granulomas. This can lead to severe complications, such as hypercalcaemia and renal failure. PMID- 8720355 TI - [Benefits and risks of immunostimulation with interleukin 12]. PMID- 8720356 TI - [Diagnosis and therapy of malignant soft-tissue tumors]. PMID- 8720357 TI - [Is iron administration, when indicated, causing any health hazard?]. PMID- 8720358 TI - [Halothane-induced hepatitis]. PMID- 8720359 TI - [Immunosuppressive therapy in an asymptomatic female patient with an autoimmune disease?]. PMID- 8720360 TI - [Interferon-alpha in chronic hepatitis C]. PMID- 8720361 TI - [Disorders of assertiveness and social anxiety]. AB - Assertiveness is a multidimensional concept use to describe individual's social functioning, from normal adaptation to pathological worries of social inefficacy. Several psychological impairments are evoked in assertiveness disorders, leading to the discussion of cognitive and emotional functioning in such pathology. These pathologies are represented by the social anxiety spectrum, an heterogeneous syndrome from shyness to social phobia in which functional dysregulation are discussed to explain the missadaptation. Experimental approach are developed to test the cognitive hypothesis proposed in assertiveness and social anxiety, and to discuss the clinical and psychopathological status of assertiveness disorders. PMID- 8720362 TI - [Levels of the training of psychotherapists and results of analytic psychotherapy]. AB - The purpose of this study was to investigate therapists experience as predictor of psychotherapy outcome. We studied consecutive patients (n = 76) assigned to psychoanalytic psychotherapy after being referred to the Geneva University Psychiatric Center for psychiatric treatment. Fifty-seven subjects were treated by trainees and nineteen had certified psychotherapists. Each treatment was carefully evaluated at intake, at the sixth session and at 1-year follow-up. Those patients treated by trainees appeared to be clearly improved (p < 0.01), but 49.1% of them did not completed the psychotherapy. In addition, increased therapist's training and experience (certified psychotherapist vs trainee) was associated with better global outcome, reduced early termination and more improvement after six psychotherapy sessions (p < 0.01). The present data indicate that increased therapist's experience is associated with more compliance and more improvement after the initial phase of the treatment in psychiatric subjects assigned to psychoanalytical psychotherapy. PMID- 8720363 TI - [Stability and predictive role of emotions, expressed by families in the development of a cohort of schizophrenic patients in the course of 5 years]. AB - A prospective study of 44 first admission schizophrenic patients was conducted in Geneva, in order to evaluate the prognostic value of Expressed Emotion (EE), as well as its stability. The predictive power of the EE index was tested on three variables of outcome: relapse rate, social adaptation and hospital stay. At intake, the patients presenting more premorbid features lived in high EE households. The EE index and the outcome measures tended to be associated. After the third year, patients living with high EE relatives were significantly more maladjusted and relapsed more than those living with low EE relatives. The EE index was stable during the five years. Our results show that initial measure of EE in a first episode cohort is predictive of outcome over a five year period. This may not be causal as it cannot be excluded that poorer premorbid functioning alone may result in poorer outcome, and may also elicit high EE in relative. PMID- 8720364 TI - [Psychological components of pregnant women who give birth to a hypotrophic child]. AB - Knowing that 20-30% of intra-uterine growth retardation are unexplained, the authors assume a psychological component that may influence fetal growth. A first psychologic approach consisted in having interviews with about fifty women who had just given birth to small-size babies. The analysis of the contents of these interviews, performed in the free association mode, allowed screening six psychopathologic themes : denial of pregnancy, painful emotional or physical experiences, operative functioning mode, unessential role of the husband, lack of an imaginary child, and previous affective deprivation. The verification of the hypothesis arising from the psychological approach will be based on a complementary epidemiological approach. PMID- 8720365 TI - [Manic-depressives, time of birth, time of onset. An epidemiological study]. PMID- 8720366 TI - [Are we really talking about psychic symptoms?]. AB - To conceive psychic symptoms as a linguistic sign stemmed from binding a form (the significant) with a meaning (the signified) assumes that rules binding signs of mental illnesses are identical to rules binding linguistic signs, but above all, that those rules are extant. The author exposes that they are not, no rules permitting to associate a linguistic form to a meaning are extant. Consequently the interpretation of psychic symptoms based on the equivalence with linguistic scheme is built on a twisted argument. PMID- 8720367 TI - [The name, scales and diagnostic criteria. Current issues]. AB - The scientific perspective incites modern psychiatry to base itself on quantification. But numbers have a double function, concrete and abstract, and this affects the meaning of statistics. Neglecting this property of each item and the associative character of the methods (scales, diagnostic criteria) will mean a risk of making the quantitative trend mythic to some degree, and may lead to a superficial, even deformed view of mental pathology. PMID- 8720368 TI - [Parapsychotic disorders in mentally handicapped (particularly in Down's syndrome)]. AB - Para psychotic troubles in Mentally Handicapped Persons (particularly in Down syndrome)-Clinical examples of M.H.P. which had no trouble until their 25-40 (years) except their intellectual defect and then, slide in psychiatric or para psychiatric behaviour. It starts with loosing vital aim and long lasting soliloquy. Then starting with an emotional frustration. Special reflexion about pseudo intellectual deterioration considered as, almost Alzheimer which is often a kind of familial contage of loosing vital impulse. It seems obvious for M.H.P. that realising their handicap without being prepared nor psychologically accompanied lead a possible trigger for loosing psychological control. Sports or leisure activities with other M.H.P. are most inefficient but a prevention often appears possible. PMID- 8720369 TI - [Document to serve the history of psychiatry. How Mr. Regis did not come to Bailleul]. PMID- 8720371 TI - Study of behaviour and endurance of Bioapatite implanted in the periodontium of the dog (closed model). AB - The principle purpose of this study was to quantify endurance of the biomaterial Bioapatite implanted in periodontal structures of the dog (closed model), such quantification being established by means of indices (Endurance Index and Transformed Endurance Index) obtained as a result of data-processed analysis of histologic images. The investigation further aimed at studying the development of new cementum and the reconstruction of an attachment system. The study was conducted on eight dogs and 222 sections. New cementogenesis and the reconstruction of an attachment system are observed both in the test sites and the reference sites. Endurance of the biomaterial is statistically linked with time: the most substantial decrease in the endurance is observed between two and six months. Traces of the material subsist at month 9. The structure of material masses always remains lacunal on a microscopic scale (highest average Endurance Index observed during the investigation: 30.35%). An osteoid deposit can be continually detected as of the second month around crystalline deposits. Further, this type of deposit was noted on the periphery of the deposits implanted in ectopic position in the supracrestal connective tissue. PMID- 8720372 TI - [The occlusal plane: an architectural myth or a proprioceptive myth?]. AB - From two radiographic pictures of the head in profile, in meantime of few years, with the same person, the author proves that occlusal plane doesn't vary during growth in comparison with the pterygo-clivien compass (Stable zone leasing against pterygoid process and superior clivus). The orientation of occlusal plane crossing by contact of the first molars and the middle of incisive intersection has an approximately analogous value. This value is equal to 110 degrees for the sample population, whatever the person's morphology and the lapsed time is. The occlusal plane moves parallel to himself and gets lower on a sagittal teleradiographic picture. PMID- 8720373 TI - [Polyunsaturated omega-3 fatty acids in the treatment of experimental human gingivitis]. AB - The anti-inflammatory effect of omega-3 polyunsaturated fatty acids (n-3 PUFA) has already been demonstrated in an animal model. The aim of this randomized, double-blind, versus placebo study is to assess this action on experimental gingivitis in humans. Over a 14-day period (day 0-day 14), 37 healthy volunteers undertook intensive oral hygiene, and then did not brush their teeth for 21 days (day 14-day 35) so that gingivitis could then develop. On day 28, the subjects were randomized in two groups: 18 in the treatment group (fish oil: 1.8 g of n-3 PUFA), 19 in the placebo group (olive oil), at a daily dose of 6 g over days (day 28-day 35). The Plaque Index (PI), the Gingival Index (GI) and the Papilla Bleeding Index (PBI), as inflammation markers, were measured on day 14, day 28 and day 35. On day 28 and day 35, five volunteers of each group underwent removal of an interdental papilla to carry out the n-3 PUFA composition of cell membranes: arachidonic acid (AA), eicosapentaenoic acid (EPA), docosapentaenoic acid (DPA), docosahexaenoic acid (DHA). The results show the integration of EPA, DHA and DPA in the membranes of the cells removed and particularly a significant increase of EPA in the treatment group (p = 0.04 S). GI in the treatment group decreased significantly (p = 0.008 S). The level of AA decrease, but no significantly. It would therefore seem that the n-3 PUFA have an effect on the reduction of gingival inflammation in this experimental gingival model in humans. PMID- 8720375 TI - Ultrastructural aspects of two different mast cell populations in human healthy gingival tissue. AB - The results of our recent microscopy studies clearly have demonstrated the constant presence of numerous metachromatic cells in healthy human gingival connective tissue. Despite the great number of studies on mast cell population in many human organs (lung, skin, uterus, and bowel), at the present time few are the studies regarding the morphostructural aspects of mast cells in the human gingiva. The aim of this study was to assess by transmission electron microscopy the presence of mast cells in the healthy human gingiva and to characterize the ultrastructural aspects of mast cells populations. 30 specimens of human gingival tissue were collected from 30 patients with informed consent. The samples were prepared for T.E.M. examination. In all the ultrathin sections observed we detected numerous and ubiquitarious mast cells. These exhibited several morphological types of cytoplasmic granules with characteristic subgranular architectural variety in shape and density. This allowed us to divide mast cells into two groups: cells with granules consisted of compact coiled scrolls, fine granular material and lattice--grafting configuration, and cells containing granules with discrete scrolls formed by more concentric lamellae and particulate structure. The two ultrastructural aspects observed correspond to McTC and McT of the international literature. Therefore in the human gingival connective tissue, like in other organs, two types of mast cells are clearly present. Surprisingly, the human gingival tissue shows, like the lung, McT as the prevailing subpopulation, in contrast to the skin, uterus and gastrointestinal submucosa where McTC prevail. PMID- 8720374 TI - Phenotypic analysis of peripheral blood cell immunity in Italian patients with different varieties of oral lichen planus. AB - Quantitative analysis of peripheral blood lymphocytes was carried out in 25 patients with atrophic-erosive type of oral lichen planus (OLP) (Group 1), in 28 patients with reticular-plaque like lesions of OLP (Group 2) and in 21 healthy patients (Group 3) by using flow cytometry. CD4+ subsets decreased significantly in patients with reticular-plaque like varieties when compared with healthy patients (Group 3) (One way analysis of variance p = 0.039; t-test with Bonferroni correction p < 0.05). Moreover, in patients with hyperkeratosic forms of OLP (Group 2) CD8+ cell populations were significantly higher than in controls (Group 3) (Kruskal-Wallis test p = 0.035; Mann-Whitney test with Bonferroni's correction p < 0.0001) and consequently CD4/CD8 ratio was significantly lower in patients with reticular-plaque like lesions than in controls (Kruskal-Wallis test p = 0.01; Mann-Whitney test with Bonferroni's correction p = 0.013). No statistical differences between patients of Group 1 (atrophic-erosive OLP) and the other two Groups (hyperkeratosic OLP and healthy controls) were detected. 40% of the patients of Group 1 were affected by chronic hepatopathies, most of which were related to hepatitis C virus (HCV), but the data were not substantially modified after adjustment for the patients with chronic liver disease HCV positive. There is no clear evidence that these results indicate the existence of a different pathogenetic mechanism between erosive-atrophic and hyperkeratosic types of OLP. On the other hand, these results and the previously reported immunohistochemical findings suggest that quantitative alterations of peripheral blood lymphocytes in hyperkeratosic varieties of OLP could represent a shift of CD4+ cells from the vascular to the oral mucosa compartment. PMID- 8720376 TI - The coronoid process as a new donor source for autogenous bone grafts for reconstructing orbital and midface defects. AB - Reconstructing orbital or midface defects by means of autogenous bone graft is a question of a well suited donor area. Several approaches in harvesting bone grafts from the calvarian or the illiac crest demonstrated that the grafts did not fit in shape or thickness. Also, the retromolar region as a donor area lead to structural loss of stability with the risk of spontaneous fractures of the mandible. Therefore, we present a new technique of harvesting an autogenous bone graft from the lateral cortical layer of the coronoid process. This region is a well suited donor area because of its easy accessibility, the good shape and thickness of the graft and its corticocancellous nature. PMID- 8720378 TI - Effect of jinkoh-eremol and agarospirol from agarwood on the central nervous system in mice. AB - Agarwood (Jinkoh in Japanese), one of the Oriental medicines, is used as a sedative. The benzene extract of this medicine showed a prolonged effect on the hexobarbital-induced sleeping time, and hypothermic effects in terms of rectal temperature, a suppressive effect on acetic acid-writhing, and a reduction of the spontaneous motility in mice. By repeated fractionation, oral administration in mice, and pharmacological screening, the active principles, jinkoh-eremol and agarospirol, were obtained from the benzene extract. They also gave positive effects on the central nervous system by peritoneal and intracerebroventricular administration. They decreased both methamphetamine- and apomorphine-induced spontaneous motility. The level of homovanillic acid in the brain was increased by them, while the levels of monoamines and other metabolites were unchanged. Similar results were seen in chlorpromazine-administered mice. Therefore, jinkoh eremol and agarospirol can be considered to be neuroleptic. PMID- 8720377 TI - Influence of a hypoiodite mouth-wash on dental plaque formation in vivo. AB - This study describes an in vivo inhibition of dental plaque growth after peroxidase-generated hypoiodite (OI-) mouth-washes. After giving up all other usual hygiene procedures nine healthy volunteers washed their mouth using 10 ml of the mouth-wash [H2O2 (0.005%), KI (50 mM) and lactoperoxidase (0.04%)] three times a day for 1 minute for 3 days. The initial oxidation power of this mixture represented 430 +/- 11 microM oxidised cysteine (n = 6), dropping down to 87 +/- 6 microM after the solution was spat out (n = 5). A saline solution served as a negative control, and a 0.2% chlorhexidine digluconate solution as a positive control. Proximal dental plaque between mandibular canine and lateral incisor (left and right) was collected after 3 days using standardized sterile toothpicks, then analysed for ATP and protein content. ATP concentrations dropped to 49% of the control values after OI- rinsing, and to 9% after chlorhexidine rinsing while the protein content dropped to 48% for OI- versus 31% for chlorhexidine. However, when considering the ATP content per protein micrograms, only the decrease to 6% of the initial value in the chlorhexidine testing was significant while the drop to 81% for the OI- testings was not significant. This study points out a negative effect of OI- on plaque growth in vivo. PMID- 8720379 TI - Decursin: a cytotoxic agent and protein kinase C activator from the root of Angelica gigas. AB - A cytotoxic compound was purified from the root of Angelica gigas Nakai by silica gel chromatography and preparative HPLC. As a result of the structure analysis by mass, IR, 1H-NMR, and 13C-NMR spectrometry, the effective compound was identified as decursin, a pyranocoumarin characterized originally from Angelica decursiva Fr. et Sav. In vitro cytotoxicity testing showed that decursin displayed toxic activity against various human cancer cell lines, for which the ED50 of decursin was about 5-16 micrograms/ml. On the other hand, decursin displayed relatively low cytotoxicity against normal fibroblasts. Decursin also activated protein kinase C (PKC) in vitro, which indicates that the cytotoxic activity of decursin may be related to the protein kinase C activation. PMID- 8720381 TI - Pectic polysaccharides from roots of Glycyrrhiza uralensis: possible contribution of neutral oligosaccharides in the galacturonase-resistant region to anti complementary and mitogenic activities. AB - Digestion with endo-alpha-(1-->4)-polygalacturonase liberated the enzyme resistant region (PG-1c) as an active site of the anti-complementary and mitogenic pectic polysaccharide (GR-2IIc) from Glycyrrhiza uralensis. Partial acid hydrolysis of PG-1c resulted in acidic oligosaccharides, and methylation analysis and GC-MS analysis of the acidic oligosaccharides suggested that PG-1c comprised a rhamnogalacturonan core such as -->2)-Rha-(1-->4)-GalA-(1-->2)-Rha-(1 ->4)-GalA-(1-->-->4)-GalA-(1-->4) as the acidic moiety. Degradation of uronic acids by lithium decreased the anti-complementary and mitogenic activities of PG 1c. Although the products from PG-1c were still active, the methylglycoside of alpha-L-Rha-(1-->4)-alpha-D-GalA-(1-->2)-alpha-L-Rha-(1-->4)-alpha-D-Gal A did not show both activities. The products obtained by the lithium degradation from PG-1c gave fractions containing various neutral oligosaccharide-alditols. Among these fractions the longest and the short oligosaccharide-alditol fractions had relatively potent anti-complementary activity, whereas all oligosaccharide alditol fractions expressed weak but significant mitogenic activity. GC-MS analysis indicated that the short oligosaccharide-alditol fraction contained various kinds of di- to tetrasaccharide-alditols. However, malto-oligosaccharide alditols, and malto-, isomalto-, and laminari-oligosaccharides did not show anti complementary and/or mitogenic activities, and these results suggested that certain neutral carbohydrate chains in PG-1c were responsible for the expression of mitogenic activity as well as anti-complementary activity of PG-1c. PMID- 8720380 TI - Anti-complementary activity of Crataegus sinaica. AB - The 80% and 70% acetone extracts from fruits and leaves of Crataegus sinaica Boiss (Rosaceae) and the ethyl acetate-, butanol-, and water-fractions obtained from these initial extracts as well as the isolated compounds, quercetin (1) (1), hyperoside (quercetin 3-O-galactoside) (2) (2), rutin (quercetin 3-O-rutinoside) (3) (2), vitexin (4) (1), rhamnosylvitexin (5) (3), monoacetylrhamnosylvitexin (6) (3), epicatechin (7) (4), procyanidin B-5 (8) (4), proanthocyanidin A-2 (9) (5), procyanidin B-2 (10) (4), and procyanidin C-1 (11) (4), were tested for their influence on the classical (CP) and alternative (AP) pathways of complement mediated hemolysis. All extracts and fractions showed a strong anti-complementary effect in a dose-dependent way which was more pronounced on the CP than on the AP. The results indicated that the pure proanthocyanidins were active on the CP. Procyanidin C-1 and proanthocyanidin A-2 were the most active on the CP and also showed activity on AP, whereas the flavonoids isolated were generally less active. However, rutin, showed a strong activity, quercetin and rhamnosylvitexin a moderate activity on the CP. This is the first report on the chemical constituents and complement-modulating activity of C. sinaica and on the occurrence of proanthocyanidin A-2 (9) in hawthorn. PMID- 8720382 TI - Inhibition of adenosine deaminase activity of aortic endothelial cells by selected flavonoids. AB - Flavonols like quercetin, myricetin, and kaempferol inhibit the activity of the adenosine deaminase of endothelial cells (IC50 between 26 and 32 mumol/l), while flavones are inactive. These results support the suggestion that many pharmacological actions of flavonoids are mediated by an amplification of the effect of endogenous adenosine via adenosine receptors because adenosine deaminase is responsible for the adenosine inactivation. PMID- 8720383 TI - In vitro biological activities of alkaloids from Cryptolepis sanguinolenta. AB - In our biological screening of higher plants, an aqueous and an 80% EtOH extract from the root bark of Cryptolepis sanguinolenta showed potent antibacterial, anticomplementary, and moderate antiviral activities, but no antifungal effect could be detected. Bioassay-guided fractionation of the 80% EtOH extract led to the isolation of three alkaloids: quindoline (1), hydroxycryptolepine (2), cryptolepine.HCl (3), and the corresponding base cryptolepine (4). All compounds strongly inhibited the growth of Gram-positive bacteria (MIC < or = 100 micrograms/ml) and showed a moderate (MIC = 125 or 250 micrograms/ml), a weak (MIC = 500 micrograms/ml), or no activity (MIC > 500 micrograms/ml) against selected Gram-negative bacteria. They also possessed a bactericidal effect depending on the bacterial strain. Compounds 1, 2 and 3 displayed a dose dependent inhibitory effect on the classical pathway of the complement system while compounds 2 and 3 activated the alternative pathway, except for compound 1. Compound 3 was found to possess an antiherpetic activity. Compounds 1 and 4 showed no antiviral effect, but were quite cytotoxic in the antiviral test system down to a concentration of 1 microgram/ml. PMID- 8720384 TI - Naturally occurring somatostatin and vasoactive intestinal peptide inhibitors. Isolation of alkaloids from two marine sponges. AB - The vasoactive intestinal peptide (VIP) and somatostatin (somatotropin release inhibiting factor, SRIF) are important neurotransmitters in a number of basic physiological events. Their disturbances have been reported in many diseases such as cystic fibrosis, impotent man (VIP), Alzheimer's disease, and some tumours (SRIF). Xestospongine B (1), sceptrine (2), and ageliferine (3), three alkaloids isolated from Xestospongia sp. and Agelas novaecaledoniae are reported as somatostatin and VIP inhibitors. The natural products 1, 2 and 3 exhibited a high affinity for somatostatin (IC50 = 12 microM, 0.27 microM, and 2.2 microM, respectively), 2 and 3 showed an affinity for VIP (19.8 microM and 19.2 microM, respectively). Due to the interaction between non-peptidic compounds and somatostatin/VIP receptors, these three alkaloids could be promising agents in the research on natural non-peptidic compounds for therapeutical interventions. PMID- 8720385 TI - Inhibitory effects of protoberberine alkaloids from the roots of Coptis japonica on catecholamine biosynthesis in PC12 cells. AB - The effects of protoberberine alkaloids from Coptis japonica Makino (COPT) on catecholamine content and tyrosine hydroxylase (TH) activity in PC12 cells were investigated. The butanol (BuOH) fraction from COPT at a concentration of 40 micrograms/ml medium inhibited catecholamine biosynthesis. The BuOH fraction was further fractionated into separate bioactive fractions (Fr. I-IV) by silica gel column chromatography, and Fr. II and III, which contain protoberberine alkaloids such as berberine and palmatine, were isolated. Fr. III at 40 micrograms/ml medium showed 77% inhibition on dopamine content in PC12 cells (IC50; 19.5 micrograms/ml medium). TH activity was reduced by the treatment of Fr. III. Berberine and palmatine also showed an inhibitory activity on dopamine biosynthesis in PC12 cells. The IC50 values of berberine and palmatine were 9.5 and 7.7 micrograms/ml medium, respectively. This suggests that the protoberberine alkaloids from COPT have an inhibitory effect on catecholamine biosynthesis by reducing TH activity in PC12 cells. PMID- 8720386 TI - Secoiridoid glucosides from Swertia mileensis. AB - Five new acyl secoiridoid glucosides along with three known secoiridoid glucosides, swertiamarin, 2'-O-acetylswertiamarin, and amarogentin, were isolated from the aerial parts of Swertia mileensis. The structures of these compounds were determined to be 2'-O-acetyl-4'-O-trans-feruloylswertiamarin, 2'-O-acetyl-4' O-cis-feruloylswertiamarin, 2'-O-acetyl-4'-O-trans-p-coumaroylswertiamarin, 2'-O acetyl-4'-O-cis-p-coumaroylswertiamarin, and 4'-O-trans-p-coumaroylswertiamarin from spectroscopic evidence. PMID- 8720387 TI - Cyclogossine A: a novel cyclic heptapeptide isolated from the latex of Jatropha gossypifolia. AB - From the latex of Jatropha gossypifolia L. (Euphorbiaceae) a novel cyclic heptapeptide was isolated, which we named cyclogossine A. A combination of amino acid analysis, FAB mass spectrometry, and two dimensional 1H-NMR spectroscopy (TOCSY and ROESY) was used to determine the primary structure. The compound was found to contain one glycine, one alanine, one valine, two leucine, one threonine, and one tryptophan residue; its amino acid sequence is: Leu 1 - Ala 2 Thr 3 - Trp 4 - Leu 5 - Gly 6 - Val 7. The absolute configurations of the amino acids were determined by chiral gas chromatography; all have the L-configuration. PMID- 8720388 TI - Production of hypericin, pseudohypericin and flavonoids in cell cultures of various Hypericum species and their chemotypes. AB - Suspension cultures were established from the shoots of sterile germinated seeds of various provenances of seven Hypericum (H.) species in a half strength modified Murashige and Skoog liquid medium. In most strains of H. perforatum (18 provenances) and all strains of H. maculatum (6 provenances) as well as in the cultures of H. tomentosum, H. bithynicum, H. glandulosum and H. balearicum, hypericin and pseudohypericin could be proven, however, in extremely varying amounts. In general, the pseudohypericin content was significantly higher than that of hypericin. The flavonoid patterns, comprising monomeric quercetin derivatives and dimeric apigenin derivatives, varied among strains over a wide range. PMID- 8720389 TI - Effects of some components of the essential oil of chamomile, Chamomilla recutita, on histamine release from rat mast cells. AB - The influence of three compounds of the essential oil of Chamomilla recutita (L.) Rauschert on the protamine sulphate-provoked degranulation of mast cells from Lewis-1a rats was investigated. The effect was determined by measuring histamine liberation fluorometrically. Chamazulene and (-)-alpha-bisabolol had no distinct effects. The en-yne dicycloether partly inhibited the degranulation in concentrations above 10(-4) M. PMID- 8720390 TI - Assessment of the anti-HIV activity of a pine cone isolate. AB - Characterization of an alkali-soluble isolate from Pinus nigra Arnold (Pinaceae) seed cones by thioacidolysis revealed a significant phenylpropanoid (i.c. lignin) component which may provide the observed anti-HIV-1 activity for this substance. PMID- 8720391 TI - An antibacterial thiophene from Balsamorhiza sagittata. AB - Balsamorhiza sagittata, a species of ethnopharmacological interest in British Columbia, is reported to have antibacterial and antifungal properties. An antibacterial compound isolated from this species was identified as 7,10-epithio 7,9-tridecadiene-3,5,11-triyne-1,2-diol based on the HMQC and HMBC experiments. PMID- 8720392 TI - Sesquiterpenes with antibacterial activity from Epaltes mexicana. PMID- 8720393 TI - Antifungal and antibacterial chalcones from Myrica serrata. AB - The dichloromethane extract of the leaves of Myrica serrata inhibits growth of Cladosporium cucumerinum, Bacillus subtilis, and Escherichia coli on TLC plates. Activity-guided fractionation led the isolation of 2',4'-dihydroxy-6'-methoxy 3',5'-dimethylchalcone (1), 2',4'-dihydroxy-6'-methoxy-5'-methylchalcone (aurentiacin A) (2), 2',6'-dihydroxy-4'-methoxy-3',5'-dimethyldihydrochalcone (3), 2'-hydroxy-4',6'-dimethoxy-3'-methyldihydrochalcone (4), and 2', 6' dihydroxy-4'-methoxy-3'-methyldihydrochalcone (5). In addition, the flavanones demethoxymatteucinol (6) and cryptostrobin (7) were also identified. PMID- 8720394 TI - Ginsenoside Rh4, a genuine dammarane glycoside from Korean red ginseng. AB - A genuine glycoside, named ginsenoside Rh4, was isolated from Korean red ginseng (Panax ginseng C. A. Meyer) through repeated column chromatography, and its chemical structure was established to be 6-O-beta-D-glucopyranosyldammar 20(22),24-diene-3 beta,6 alpha,12 beta-triol by spectral and chemical methods. The stereochemistry of a double bond at C-20(22) of ginsenoside Rh4 was characterized as (E) from a NOESY experiment in the 1H-NMR of the aglycone. Cytotoxic activities of ginsenoside Rh4 and its aglycone against cancer cell lines were evaluated by use of the SRB method. PMID- 8720395 TI - 1995 Inaugural address. PMID- 8720396 TI - Gender differences in medical school attrition rates, 1973-1992. AB - Retention is a critical problem in medical school education. We report here on research that examined gender differences in attrition rates between 1973 and 1992. Using secondary data compiled from the annual reports on undergraduate education published in JAMA, both descriptive and inferential analyses of medical school attrition rates were conducted. Data show that medical school attrition rates have steadily increased across the country since 1973 and that women drop out of medical school at consistently greater rates than men. These results highlight the importance of future analyses that attempt to delineate the causes as well as the consequences of dropping out of medical school for women and the institutions that support them. PMID- 8720397 TI - Harassment of women physicians. AB - This paper reviews current knowledge about the prevalence, characteristics, and costs of sexual harassment of women medical students and physicians. It also addresses the limited research on other forms of physician and student harassment, and notes the kinds of information that are still needed. PMID- 8720398 TI - Experimental chemonucleolysis with chondroitinase ABC in monkeys. AB - STUDY DESIGN: Experimental chemonucleolysis with chondroitinase ABC as compared with chymopapain was investigated in monkeys. The effects of these two enzymes were analyzed morphologically and biochemically. OBJECTIVES: The results of the present study facilitate the clinical application of chondroitinase ABC. SUMMARY OF BACKGROUND DATA: Many experimental studies on chemonucleolysis with chondroitinase ABC have been reported mostly in rabbits. Further examination in animals that are more similar to humans is necessary for its clinical application. METHODS: Ten mature rhesus monkeys were used. Lumbar intervertebral discs from six monkeys were injected with either chondroitinase ABC, chymopapain, or physiologic saline. The animals were killed not more than 6 weeks after injection, and their discs were examined morphologically and biochemically. The remaining four monkeys were used for a longer follow-up study to determine the changes in chondroitinase ABC-injected discs. RESULTS: Degenerative changes of discs were more severe with the injection of chymopapain, although the content of hyaluronic acid, chondroitin sulfate, and dermatan sulfate decreased more with the injection of chondroitinase ABC. In a longer follow-up study, chondroitinase ABC-injected discs recovered somewhat morphologically and biochemically. CONCLUSIONS: The results confirm that selective degradation is achieved with chondroitinase ABC in vivo in monkeys and that chondroitinase ABC is less toxic to discs than chymopapain is. From these considerations, chondroitinase ABC is believed to have a high potential for clinical application. PMID- 8720399 TI - Experimental acute dorsal compression of cat spinal cord: correlation of magnetic resonance signal intensity with spinal cord evoked potentials and morphology. AB - STUDY DESIGN: Acute dorsal compression of the spinal cord was applied to adult cats, and magnetic resonance signal intensity, spinal cord evoked potentials, and morphologic changes of the spinal cord were examined after 5 hours. OBJECTIVES: The present study investigated the correlation of magnetic resonance signal intensity with spinal cord evoked potentials and spinal cord morphology after 5 hours of spinal cord compression in cats. SUMMARY OF BACKGROUND DATA: Neurologic prognosis of the injury might be predicted by an analysis of magnetic resonance signal intensity pattern. Little information is available on relationships between magnetic resonance images and functional or morphologic damage of spinal cord in acute animal experiments. METHODS: Acute dorsal compression of the spinal cord was performed in 24 anesthetized cats. After laminectomy, the L2 segment was compressed for 5 hours. Spinal cord evoked potentials were recorded by electrodes placed in the epidural space at L4, and the spinal cord was stimulated at T12. The animals were divided into four groups based on changes in the amplitude of spinal cord evoked potentials. Immediately after compression for 5 hours, magnetic resonance images were obtained. Signal intensity of the spinal cord was measured on sagittal midline images. Morphologic changes were assessed. RESULTS: Spinal compression significantly increased the signal intensity of the L1, L2, and L3 segments on T2-weighted and proton density-weighted images. The increase in signal intensity was remarkable in the animals whose spinal cord evoked potentials were reduced greatly (< 40% of the control group). Histologically, edema was present in the high intensity area on T2-weighted and proton density weighted images. CONCLUSIONS: In summary, the present study documents that spinal compression causes tissue edema, which produces high signal intensity on magnetic resonance imaging. The magnetic resonance signal intensity is correlated closely with decreased amplitude of spinal cord evoked potentials. PMID- 8720400 TI - Morphologic differences of the vascular buds in the vertebral endplate: scanning electron microscopic study. AB - STUDY DESIGN: Vascular buds in rabbit vertebral endplates were examined by scanning electron microscopy of corrosion casts. OBJECTIVES: To examine morphologic differences between vascular buds in two regions of the vertebral endplate (inner anular and nucleus pulposar). SUMMARY OF BACKGROUND DATA: Vascular buds are specific structures present at the vertebral endplate that are important as nourishing channels. There is a significant difference in permeability between the lateral portion (inner anular) and the central portion (nucleus pulposar) of the endplate, the latter usually being permeable and the former being impermeable. Morphologic differences between vascular buds in the two regions have not been investigated previously. METHODS: Eight 20-week-old rabbits were used. Vascular buds in rabbit vertebral endplates were examined by scanning electron microscopy of corrosion casts. RESULTS: The vascular buds in the region of the inner anulus form simple loops, but those in the area near the nucleus pulposus exhibit swollen and complex coil-like loops. Although they differ structurally, the average number of vascular buds per area does not vary between the two regions. CONCLUSIONS: We suggest that the morphologic difference between the vascular buds in the two regions (inner anular and nucleus pulposar) plays a principal role in permeability at the endplate. PMID- 8720401 TI - Allografting intervertebral discs in dogs: a possible clinical application. AB - STUDY DESIGN: A lumbar disc is a tissue that is often a cause of low back pain. We conducted experimental allografting of intervertebral discs in dogs. OBJECTIVES: To evaluate the clinical potential for allografting of intervertebral disc. SUMMARY OF BACKGROUND DATA: There have been few reports on the allografting of intervertebral discs. The allografting of discs frozen at -196 C has never been documented. In the present experiments, we studied the effect of freezing at -80 C and -196 C on disc allografts to 25 hybrid adult dogs. METHODS: A vertebral disc was obtained from donor dogs with parts of the adjoining vertebral bodies to serve as the allograft disc unit. After soaking in 10% dimethyl sulfoxide at 4 C, the allograft discs were frozen and stored at either -80 C or -196 C. After storage for approximately 4 weeks, they were transplanted peritoneally into the recipient lumbar vertebrae and fixed using plates and screws to preserve mobility of the grafted disc. At the same time, cells from allograft discs were incubated to examine their ability to synthesize proteoglycan and collagen. Radiologic examination of the changes after surgery in the intervertebral space was followed serially. RESULTS: Radiographs revealed a complete bone union of the vertebral bodies at 5 months and gradual narrowing of the intervertebral space beginning at 6 months. Histologically, the anulus fibrosus was well preserved, especially in disc units stored at -196 C. Cellular synthetic activity was seriously diminished. CONCLUSIONS: Our results indicate that allograft of intervertebral discs has clinical potential because it can serve as a dynamic disc spacer for a certain period of time, but its long-term merits and demerits have to be established. PMID- 8720402 TI - Effects of strain distribution in the intervertebral discs on the progression of ossification of the posterior longitudinal ligaments. AB - STUDY DESIGN: Strain distribution in the intervertebral discs was evaluated biomechanically using an engineering true strain calculation formula. OBJECTIVES: This study was performed to clarify the involvement of dynamic factors in the progression of ossification of the posterior longitudinal ligament. SUMMARY OF BACKGROUND DATA: In patients with ossification of the posterior longitudinal ligament of the cervical spine, ossification frequently progresses after laminectomy. This suggests the involvement of dynamic factors in the progression of ossification. However, these factors have not yet been clarified. METHODS: The analysis was performed on 101 patients with ossification of the posterior longitudinal ligament by employing dynamic lateral x-ray films of the cervical spine. The x-ray films were digitized and used as computer data for calculating the strain distribution. X-ray films were obtained again 5 years later, and the strain distribution and the presence or absence of progression of ossification were evaluated. RESULTS: The progression of ossification of the posterior longitudinal ligament was highly correlated with abnormal strain distribution in the intervertebral discs. Progression of ossification was frequently observed in areas having disc distortion in tension and extension on the posterior longitudinal ligament. CONCLUSIONS: In this study, the area of progression of ossification corresponded to the area showing uneven strain distribution and resultant concentration of dynamic stress. These results suggest an important role for dynamic factors in the progression of ossification of the posterior longitudinal ligament. PMID- 8720403 TI - Cartilaginous endplate in cervical disc herniation. AB - STUDY DESIGN: Cervical herniated disc tissue obtained at surgery for myelopathy and intervertebral discs from autopsy cases were examined histologically. OBJECTIVES: To clarify the characteristic histology of cervical disc herniation and the processes by which herniated masses are produced. SUMMARY OF BACKGROUND DATA: Except for nucleus pulposus or anulus fibrosus, no other disc tissue has been described histologically in the cervical spine. METHODS: Twenty-one herniated cervical discs from 20 surgical cases (patients aged 37-68 years) and mid-sagittal slabs of 135 cervical discs from 41 autopsy cases (aged 20-85 years) were examined histologically. RESULTS: All the surgical specimens had cartilaginous endplate fragments together with nucleus pulposus or anulus fibrosus. Of the autopsy disc specimens, 61% had a horizontal cleft longer than two thirds of the anteroposterior diameter of the disc, and 49% had one or more vertical clefts extending to the cartilaginous endplate. Thirty-three percent showed separation of the cartilaginous endplate, and 15% had a herniated mass including parts of the cartilaginous endplate. These abnormal features increased with aging. The same order of decreasing frequency from horizontal cleft formation through herniation was observed in all decades. CONCLUSIONS: The cartilaginous endplate-type of herniation is the predominant type of herniation in the cervical spine. It results from horizontal and vertical cleft formations of the disc. PMID- 8720404 TI - Laminoplasty with foraminotomy for coexisting cervical myelopathy and unilateral radiculopathy: a preliminary report. AB - STUDY DESIGN: An assessment was made of the efficacy of a combined laminoplasty and foraminotomy operation for patients with coexisting myelopathy and unilateral radiculopathy. The procedure was done in 17 patients. OBJECTIVES: The patients were followed with lateral flexion and extension radiographs, computed tomography scans, and an assessment system specially designed to qualitatively evaluate the patients' neurologic status. Follow-up period averaged 4 years (range, 2.1-9.3 years). SUMMARY OF BACKGROUND DATA: Excellent-to-good results were obtained for 76% (13 of 17) of the patients without any significant functional compromise based on the radiographs. Sixteen nerve roots were decompressed with a less than 25% foraminotomy, whereas eight were decompressed by a 25%-50% foraminotomy without serious neurologic damage, except for one patient. The neurologic results appeared unrelated to the extent of foraminotomy. METHODS: A refined procedure for combined laminoplasty and foraminotomy was reviewed retrospectively in terms of neurologic outcome and radiographic data. RESULTS: The present series is small, and results are not comparable directly with other methods. The procedure appears effective for myelopathy and radiculopathy. This procedure is applicable to patients with myelopathy and coexisting nerve root impingement anterolaterally or in the neural foramen. CONCLUSION: The combined laminoplasty and foraminotomy operation may provide greater neurologic improvement in patients with coexisting myelopathy and unilateral radiculopathy, while maintaining cervical spine stability after surgery. PMID- 8720405 TI - Extradural tethering effect as one mechanism of radiculopathy complicating posterior decompression of the cervical spinal cord. AB - STUDY DESIGN: This study used anatomical and clinical data to analyze the postoperative tension-status of cervical roots after posterior decompression of the cervical spinal cord. The efficacy of longitudinal durotomy with regard to prevention of postoperative palsy was investigated in a controlled study. OBJECTIVES: To elucidate the mechanism of postoperative radiculopathy and to prevent its occurrence. SUMMARY OF BACKGROUND DATA: Previous anatomical studies by the authors revealed that the posteromedial shift of the dura-root junction following posterior bulging of the cervical dural sac exerted a traction force on the portion of the roots outside the dural sac and reduced the tension on the rootlets inside the dural sac. These traction-related phenomena disappear after longitudinal durotomy. METHODS: Lengths of various parts of an anterior root were measured in 20 Japanese adult cadavers. The shortest pre- and postoperative distances between the anterolateral mid-edge of the spinal cord and dura-root junction were compared, using computed tomography-myelograms of postoperative C5 radiculopathies. The effects of longitudinal durotomy were also investigated in a controlled study involving 118 patients with laminoplasty. RESULTS: The length ratio between the longest and shortest anterior rootlet in each cervical root showed fairly constant values. Clinically, the length of the shortest rootlet could be calculated using a myelogram. Radiographical findings in cases of postoperative radiculopathies suggested increased tension on roots outside the dural sac but not on rootlets inside the dural sac. Application of longitudinal durotomy in a type of lateral opening laminoplasty resulted in the disappearance of postoperative radiculopathy. CONCLUSIONS: An extradural tethering effect was suggested as one mechanism leading to postoperative radiculopathy. Durotomy may be useful in the treatment of postoperative palsy. PMID- 8720406 TI - A new method using top views of the spine to predict the progression of curves in idiopathic scoliosis during growth. AB - STUDY DESIGN: A prospective longitudinal study of 51 patients with idiopathic scoliosis using spinal stereoradiographs was performed. The top view, which was obtained from stereoscopic anteroposterior and lateral radiographs, was analyzed for predicting the progression of spinal deformity. OBJECTIVES: To show that the top view facilitates prediction of curve progression in idiopathic scoliosis at the initial examination. SUMMARY OF BACKGROUND DATA: Four progression factors were set up using the top view and were analyzed statistically for predicting progression. No previous study has assessed this concept. METHODS: Fifty-one patients with idiopathic thoracic scoliosis or combined thoracic and lumbar scoliosis were studied longitudinally. There were 24 untreated patients and 27 patients treated with braces. Four potential progression factors were evaluated using the top view: 1) the ratio of the frontal size and the sagittal size in the top view, 2) the magnitude and direction of the vector describing the plane of maximum curvature in the thoracic spine, 3) the magnitude and direction of the vector describing the plane of maximum curvature in the lumbar spine, and 4) the balance of these vectors between the thoracic and lumbar curve. All cases were classified into five groups according to these four factors. RESULTS: The probability of the progression was evaluated statistically, and the prevalence of curve progression was found in each group. The probability of progression of a scoliosis curve increased according to the increase of these four factors. No significant difference was found between Cobb angle at the initial examination and that at skeletal maturity in untreated patients with a small risk of progression. The patients with a large risk of progression and who were treated with braces showed progression of curvature despite brace treatment. CONCLUSION: The present study has evaluated factors relating to progression in scoliosis using the top view. These results may help predict the risk of progression in idiopathic scoliosis. PMID- 8720407 TI - Inflammatory cytokines in the herniated disc of the lumbar spine. AB - STUDY DESIGN: Tissues in the area of herniated lumbar discs were examined for inflammatory cytokines to elucidate the causes of sciatic pain in lumbar disc herniation. OBJECTIVES: To determine the role of inflammatory cytokines in the stimulation of sciatic pain in lumbar disc herniation. SUMMARY OF BACKGROUND DATA: It is postulated that in addition to mechanical compression of lumbar nerve roots and sensory root ganglia by herniated discs, there is a chemical stimulus to the production of sciatic leg pain. The exact mechanisms of chemical stimulation are not clearly defined. METHODS: During surgery, cases of lumbar disc herniation in 77 patients were classified macroscopically into protrusion, extrusion, and sequestration types. Tissues adjacent to nerve roots at the herniation were excised and analyzed biochemically and immunohistochemically for the presence of inflammatory cytokines and for the production of these cytokines and prostaglandin E2 in vitro. RESULTS: The homogenates of samples were analyzed for interleukin-1 alpha, interleukin-1 beta, interleukin-6, tumor necrosis factor alpha, and granulocyte-macrophage colony stimulating factor, which were detectable. Most of the cytokine-producing cells were histiocytes, fibroblasts, or endothelial cells in extrusion and sequestration types, and chondrocytes in protrusion type. The secretion of these cytokines and prostaglandin E2 was decreased by the addition of betamethasone. The prostaglandin E2 production was dramatically enhanced by additional interleukin-1 alpha, but decreased by the addition of tumor necrosis factor-alpha. CONCLUSION: The results demonstrate that at the site of lumbar disc herniation, inflammatory cytokines such as interleukin 1 alpha are produced, which increases prostaglandin E2 production. Further studies are required to elucidate the role of inflammatory cytokines in causing sciatic pain. PMID- 8720408 TI - The natural history of herniated nucleus pulposus with radiculopathy. AB - STUDY DESIGN: The present study retrospectively investigated the morphologic changes that occurred during conservative treatment of patients with unilateral leg pain resulting from herniated nucleus pulposus without significant lumbar canal stenosis. OBJECTIVES: The results were correlated with clinical outcomes and extruding forms to determine which type of herniated nucleus pulposus had the greatest capacity for spontaneous regression and how rapidly such regression might occur. SUMMARY OF BACKGROUND DATA: The study population consisted of 77 patients with radiculopathy. METHODS: All patients complained primarily of unilateral leg pain, and 94% had positive tension signs. Additionally, 32% exhibited muscle weakness corresponding to the symptomatic nerve root. All patients were studied more than twice using magnetic resonance imaging during conservative therapy at a mean interval of 150 days. Morphologic changes on magnetic resonance imaging fell into four categories, with herniated nucleus pulposus classified into three types using T1-weighted sagittal views. Each patient was reexamined on the same scanner; 53 patients were examined twice, and 24 patients were examined more than three times. RESULTS: Morphologic changes, with the exception of 13 false-negative cases, basically corresponded to clinical outcome. In half of the cases that showed some improvement at follow-up evaluation, improvement of clinical findings were seen before those observed on magnetic resonance imaging. Migrating herniated nucleus pulposus frequently presented an obvious decrease in size, and even disappearance in seven cases. The further the herniated nucleus pulposus migrated, the more decrease in size could be observed. The cases apparently corresponding to "protrusion" showed little or no change on follow-up magnetic resonance imaging. Regarding the mechanism of herniated nucleus pulposus disappearance, exposure to the vascular supply undoubtedly took a part, although many factors were suspected to have some influence. CONCLUSION: Morphologic changes on magnetic resonance imaging mainly corresponded to clinical outcomes but tended to lag behind improvement of leg pain. Disappearance of herniate nucleus pulposus was seen frequently in the cases of migrating disc herniation, and it was presumed that exposure to the vascular supply had a lot to do with this phenomenon. PMID- 8720409 TI - Histologic evidence of absorption of sequestration-type herniated disc. AB - STUDY DESIGN: The reactions to sequestrated disc fragments, which were removed surgically from 35 patients, were examined histologically. OBJECTIVES: To elucidate whether or not there is histologic evidence of absorption of sequestrated discs. SUMMARY OF BACKGROUND DATA: Spontaneous disappearance or diminution of lumbar herniated discs in the spinal canal has been recognized, and this could be a possible explanation for relief of symptoms without surgery. The mechanism of this phenomenon is unclear. METHODS: Sequestrated discs removed surgically from 35 patients were examined histologically. RESULTS: In 30 cases, neovascularization was observed at the periphery of the sequestrated discs. Many foamy cells (macrophages) were present in the vascularized areas. In addition, immunohistochemistry revealed that many spindle-shaped, fibroblast-like cells were positive for CD68, a marker of macrophages. No fibrous scar formation was observed in any region. CONCLUSION: These findings suggest that organization is not a main course for this type of herniated disc and that a kind of "absorption" process occurs predominantly in the healing stage. PMID- 8720410 TI - Immunohistologic study of the ruptured intervertebral disc of the lumbar spine. AB - STUDY DESIGN: This study analyzed immunohistological features of the extruded or sequestrated intervertebral disc of the lumbar spine. To clarify the pathogenesis of neovascularization, cells isolated from herniated disc were cultured and examined biologically. OBJECTIVES: The objectives of this study were to characterize the histologic features of extruded or sequestrated discs and inflammatory cells that infiltrate along the margins of the disc tissue and to clarify the pathogenesis of neovascularization observed at the edge of the disc tissue. SUMMARY OF BACKGROUND DATA: When some of the contents of the disc extrudes into the epidural space and is considered "foreign," an autoimmune response develops, which can lead to a chronic inflammatory response. However, the pathogenesis of inflammatory cell infiltrations and neovascularization are not clearly defined. METHODS: The herniated discs were obtained during surgery and were stained with anti-interleukin-1, intercellular adhesion molecule-1, lymphocyte function-associated antigen, and basic fibroblast growth factor antibodies by using an indirect immunoperoxidase method. Cells isolated from herniated disc were cocultured with human endothelial cells and basic fibroblast growth factor contained by cultured disc cells were measured by radioimmunoassay. RESULTS: The ingrowth of granulation tissue with vascularization, occurring at the edge of fibrocartilage fragment, was present at 11 of 16 of extruded and 3 of 5 of sequestrated discs. Anti-interleukin-1, intracellular adhesion molecule-1, lymphocyte function-associated antigen, and basic fibroblast growth factor were expressed on most of mononuclear cells infiltrating into the extruded or sequestrated disc. Cells from the extruded or sequestrated disc demonstrated significantly greater levels of basic fibroblast growth factor than those from the protruded disc, and they enhanced the proliferation of endothelial cells. CONCLUSIONS: This study showed that mononuclear cells infiltrating along the margins of extruded discs expressed inflammatory mediators and might induce neovascularization and persistence of inflammation. PMID- 8720411 TI - Indication of fusion for lumbar spinal stenosis in elderly patients and its significance. AB - STUDY DESIGN: Selection of surgical therapy for lumbar canal stenosis in elderly patients is discussed. Decompression alone and decompression with fusion were evaluated. OBJECTIVES: To determine the indication of decompression with fusion for lumbar spinal stenosis in elderly patients. SUMMARY OF BACKGROUND DATA: Although there is no objection to posterior decompression, which is regarded as the first choice of surgical therapy for lumbar spinal stenosis in the elderly, it is debatable whether or not fusion should be used with decompression. METHODS: The presence or absence of instability was defined by Posner's method from preoperative plain radiographic lateral findings. Thirty-four elderly patients with lumbar canal stenosis were studied. Seventeen of the 34 patients were found to have instability. Ten of the 17 patients with spinal instability underwent decompression and instrumented fusion. The seven remaining patients with spinal instability underwent decompression alone. The 17 patients without spinal instability were treated by decompression alone. Preoperative symptoms, postoperative results, and changes in radiographic findings were compared among the three groups. RESULTS: The group treated by decompression and fusion showed the best results. The group treated by decompression in the presence of instability showed the worst results by the Japanese Orthopaedic Association back scores. Good results can be obtained by decompression alone only if the patients do not have instability as defined by Posner. CONCLUSIONS: The definition of instability by Posner's method proved useful for selecting elderly patients with instability for fusion treatment. Fusion with instrumentation should be performed on elderly patients with instability after decompression. PMID- 8720412 TI - Tumoral calcinosis in the upper cervical spine: a case report. AB - STUDY DESIGN: An elderly women with tumoral calcinosis between the C1 posterior arch and the C2 lamina is reported. OBJECTIVES: To describe a rare occurrence of tumoral calcinosis in the spine and discuss its pathomechanism. SUMMARY OF BACKGROUND DATA: Tumoral calcinosis is a rare condition occurring predominantly in the juxtaarticular regions of the extremities. Involvement of the spine has been reported in only one case in the lumbar spine. The calcium phosphate crystals that have been identified are those of hydroxyapatite. METHODS: The calcified material was removed, and its crystalline phase was analyzed. RESULTS: Neck pain was relieved by removal of the calcified material. Scanning electron microscopy and energy-dispersive x-ray microanalysis revealed that the material consisted of crystals of calcium pyrophosphate dihydrate. CONCLUSIONS: Tumoral calcinosis should be included among the clinical presentations of calcium pyrophosphate dihydrate crystal deposition disease. PMID- 8720413 TI - A controlled comparison of myelography. PMID- 8720414 TI - Use of losartan to examine the role of the cardiac renin-angiotensin system in myocardial dysfunction during ischemia and reperfusion. AB - To assess the role of angiotensin II (AII) in the development of myocardial dysfunction during ischemia and reperfusion, the effects of either oral pretreatment with 1 mg/kg losartan or treatment with 4.5 mu M losartan in vitro were compared with effects measured in the respective placebo or in vitro control groups in an isolated rat working-heart model. Both groups treated with losartan showed significant improvement (p < 0.005) in functional recovery following 20 min of ischemia compared with the respective control groups. Coronary flow (CF) and cardiac output (CO) were also significantly increased during reperfusion in the drug treatment groups compared with controls (p < 0.05 to p < 0.001). The recovery of mechanical function, CO, and CF was significantly more rapid in hearts from rats treated orally with losartan than in hearts treated with losartan in vitro. As measured by 31P-nuclear magnetic resonance, the changes observed in ATP levels and in intracellular pH during ischemia and reperfusion were essentially the same under either treatment regimen. This article describes the initial observation of a significant reduction in myocardial dysfunction during reperfusion following 20 min of global ischemia in the isolated perfused heart as a result of acute AII AT1 receptor antagonism by losartan administered either directly in vitro or by oral pretreatment. PMID- 8720415 TI - Fosinopril reduces ADP-induced platelet aggregation in hypertensive patients. AB - Platelets are intimately involved in atherosclerosis, and hypertension is a known risk factor for coronary artery disease. The angiotensin-converting enzyme (ACE) inhibitors were demonstrated to reduce hypertension and attenuate atherosclerosis. Because increased platelet aggregation was shown in hypertensive patients, the effect of a new ACE inhibitor, fosinopril, on platelet aggregation was studied. Fosinopril therapy (10 mg/day for 4 weeks) in 18 male hypertensive patients showed > or = 31% reduction in ADP-induced platelet aggregation. In vitro studies showed that fosinopril had similar inhibitory effect on ADP-induced platelet aggregation. No inhibitory effect could be detected with collagen as the aggregating agent. Finally, inhibition of platelet aggregation by fosinopril was less effective in platelets derived from hypertensive patients as compared with platelets derived from normal subjects. We conclude that fosinopril possesses a significant inhibitory activity on ADP-induced platelet aggregation both in vitro and in vivo. PMID- 8720416 TI - Fosinopril improves regulation of vascular tone in mesenteric bed of diabetic rats. AB - Because diabetes mellitus leads to vascular dysfunction, we examined the microvascular endothelial and smooth muscle function in long-term diabetes and a possible influence of fosinopril treatment (10 mg/kg). We investigated isolated perfused mesenteric beds of diabetic rats (4 groups: control, control + fosinopril, diabetes, diabetes + fosinopril; diabetes of 6-month duration, induced by streptozotocin, STC) were investigated using computer-assisted microvideoangiometry. Vascular diameter of four different vascular regions [classified as conductive (G1, 303 +/- 6.5 mu m and G2, 239 +/- 6.3 mu m) and resistance (G3, 192 +/- 4.5 mu m and G4, 124 +/- 2.6 mu m) vessel generations; resting conditions, control group] were increased in diabetes by approximately 20%. However, the endothelium-dependent relaxation in response to 1 mu M acetylcholine (ACh) was reduced from 38-44% to 20-25% (diabetes mellitus) with maximal impairment in G4 vessels. This could be significantly antagonized by fosinopril treatment. Similarly, vasodilation in response to 1 mu M glyceroltrinitrate (GTN) was reduced from 50-58 to 20-30%, but was partially prevented by fosinopril (32-38%), whereas potassium chloride (KCl)-induced vasoconstriction did not show differences between the groups. Inhibition of nitric oxide (NO) synthesis by 3 mu M L-NG-nitro arginine (L-NNA) resulted in a slight vasoconstriction of all vessels (12-25%), with maximum response in G3/G4. This was not altered by disease or treatment. We conclude that (a) long-term diabetes leads to endothelial and smooth muscle dysfunction with reduced capability of vasodilation and either an impairment of NO release or a reduced smooth muscle responsiveness to and (b) a predominant impairment of NO-dependent regulation in small resistance vessels, and (c) that fosinopril treatment can at least partially prevent this vascular dysfunction. PMID- 8720417 TI - Role of calcium in angiotensin II-induced prostaglandin release and DNA synthesis in rat vascular smooth muscle cells. AB - Cellular calcium modulates enzyme activity, cell proliferation, and differentiation. In vascular smooth muscle cells (VSMC), calcium may contribute to increased vascular contractility and structural alterations in both hypertension and atherosclerosis. We investigated the role of calcium in angiotensin II (AII)-induced prostaglandin release and DNA synthesis in VSMC. Prostaglandin levels were determined by radioimmunoassay, and DNA synthesis was determined by the incorporation of [3H]thymidine. AII dose-dependently stimulated the release of prostaglandin E2 and prostaglandin I2, and this effect was synergistically enhanced by the Ca2+ ionophore A23187. Conversely, the AII response was inhibited by EGTA, a chelator of Ca2+ ions and by verapamil and nifedipine, two Ca2+ channel blockers or by incubation of the cells without exogenous Ca2+. TMB-8, an inhibitor of calcium mobilization, also strongly reduced angiotensin response. Similar results were obtained for angiotensin III (AIII) and vasopressin, two other agonists of prostaglandin production. AII- or serum-stimulated DNA synthesis was almost abolished by EGTA, whereas TMB-8, verapamil, and nifedipine had little or no effect. The production of prostaglandins triggered by angiotensins and vasopressin in VSMC is dependent on both intracellular and extracellular calcium, with calcium entering through L type Ca2+ channels. Extracellular calcium is important for AII and serum mitogenic activity, but L-type Ca2+ channels do not appear to be implicated. PMID- 8720418 TI - Preservation of left ventricular mechanical function and energy metabolism in rats after myocardial infarction by the angiotensin-converting enzyme inhibitor quinapril. AB - We tested whether angiotensin-converting enzyme (ACE) inhibitor therapy with quinapril prevents the deterioration of mechanical function and high-energy phosphate metabolism that occurs in chronically infarcted heart. Rats were subjected to ligation of the left anterior descending coronary artery (LAD) or sham operation. Four groups were studied: sham-operated rats (n = 10), rats with myocardial infarction (MI, n = 9), sham-operated quinapril-treated rats (n = 8), and infarcted quinapril-treated (n = 13) rats. Treated rats received 6 mg/kg/day of the ACE inhibitor quinapril orally, initiated 1 h after MI or sham operation. Eight weeks after LAD ligation or sham operation, hearts were isolated and buffer perfused isovolumically. High-energy phosphate metabolism and intracellular pH were continuously recorded with 31P-nuclear magnetic resonance (NMR) spectroscopy. Hearts were subjected to 15-min control, 30-min hypoxia (95% N2/5% CO2, and 30-min reoxygenation. Left ventricular developed pressure (LVDP) was reduced in infarcted hearts (58 +/- 10 vs. 98 +/- 9 mm Hg in sham, p < 0.05), and this reduction was partially prevented by quinapril (78 +/- 8 mm Hg). ATP content of residual intact myocardium after sham operation or MI was unchanged. Creatine phosphate was reduced in infarcted hearts (107 +/- 10 vs. 138 +/- 5% of control ATP, p < 0.05), and quinapril prevented this decrease (131 +/- 8%). Therefore, quinapril preserved both function and high-energy phosphate metabolism in the chronically infarcted heart. However, when hearts were subjected to acute hypoxia, susceptibility to acute metabolic stress was substantially increased in both quinapril-treated groups: ATP content at end-hypoxia was reduced to 31 +/- 7 and 37 +/- 6% in sham and infarcted quinapril-treated groups, whereas ATP in untreated sham and infarcted hearts was 66 +/- 6 and 66 +/- 3% of baseline values (p < 0.05 untreated vs. quinapril treated). Likewise, recovery of LVDP during reoxygenation was impaired by quinapril treatment (15 +/- 7 and 15 +/- 4 mm Hg in quinapril-treated sham and MI vs. 73 +/- 9 and 46 +/- 9 mm Hg in untreated sham and MI groups, p < 0.05 untreated vs. quinapril treated). The most likely explanation for the unexpected finding of increased susceptibility to acute metabolic stress in the quinapril-treated groups is reduced wall thickness leading to increased wall stress. The preservation of high-energy phosphate content in residual intact hearts after MI may contribute to the beneficial effects of ACE inhibitors after MI. PMID- 8720419 TI - Effect of sodium nitroprusside on norepinephrine overflow and antidiuresis induced by stimulation of renal nerves in anesthetized dogs. AB - To investigate the role of nitric oxide (NO) in the regulation of renal sympathetic nerve activity and renal function, we examined the effect of sodium nitroprusside (SNP), a NO donor, on renal actions induced by renal nerve stimulation (RNS) in anesthetized dogs, with or without blockade of an endogenous NO generation by NG-nitro-L-arginine (NOARG), a NO synthase inhibitor. Low frequency RNS (0.5-2.0 Hz) enhanced the rate of norepinephrine secretion rate (NESR) from the kidney and decreased urine flow (UF), urinary excretion of sodium (U(Na)V), and fractional excretion of sodium (FENa, without affecting systemic and renal hemodynamics. The intrarenal arterial infusion of SNP, in a dose (1 mu g/kg/min) that does not affect renal hemodynamics and urine formation at the basal level, significantly attenuated the RNS-induced decreases in UF, UNa V and FENa. The intrarenal administration of NOARG (40 mu g/kg/min) elicited renal vasoconstriction and reduced urine formation. RNS during NOARG administration reduced renal blood flow (RBF) and glomerular filtration rate (GFR) and augmented RNS-induced reduction in urine formation. Simultaneously, NESR was markedly enhanced. The renal actions observed with NOARG administration during control and RNS periods were almost completely abolished by treatment with SNP. Therefore, we suggest that NO plays an important role in the regulation of renal function. Endogenous NO probably functions as an inhibitory modulator of renal noradrenergic neurotransmission at the prejunctional level. PMID- 8720420 TI - Class III antiarrhythmic effects of LY-190147 on defibrillation threshold. AB - Defibrillation strength shocks delivered within an action potential (AP) delay repolarization. Shock-induced AP duration extension (APDE) may prolong refractoriness and terminate or prevent reinitiation of reentry, favoring defibrillation. This study examined LY-190147 (LY) effects on defibrillation threshold (DFT) in 11 dogs. Ventricular effective refractory period (VERP) and epicardial monophasic AP duration at 75% repolarization (APD75) were recorded at 300-, 400-, 500-, and 600-ms pacing cycle length (CL). APDE was measured as the time to 50% repolarization after a DFT strength shock delivered at 50, 25, and 0 ms before or 25 ms after VERP during pacing at 300 ms CL in 4 of the dogs. We made all recordings before drug administration and after infusions of 0.03, 0.3, and 3.0 mg/kg LY, using 1.5-h dosing intervals. LY lowered DFT in a saturating dose-response manner whether expressed as shock peak voltage (V) or energy. LY decreased DFT-V from 357 +/- 77 V before drug to 331 +/- 60 V (-6 +/- 12%), 290 +/- 43 V (-17 +/- 13%, p < 0.001), and 312 +/- 45 V (-11 +/- 12%, p < 0.05) at 0.03, 0.3, and 3.0 mg/kg, respectively. Similarly, LY treatment decreased defibrillation energy requirements from 6.9 +/- 2.7 J before drug by 7 +/- 25%, 26 +/- 24%, and 12 +/- 25% at the same doses. At 300-600 ms CL, LY prolonged APD75 by an average of 10 +/- 8% at 0.03 mg/kg, 17 +/- 6% at 0.3 mg/kg, and 24 +/ 9% at 3 mg/kg. At these CL, LY prolonged VERP by an average of 4 +/- 6% at 0.03 mg/kg, 15 +/- 10% at 0.3 mg/kg, and 11 +/- 9% at 3 mg/kg. APDE was increased from 62 +/- 9 ms before to 68 +/- 14, 80 +/- 16 (p < 0.001) and 72 +/- 13 ms (p < 0.05) at 0.03, 0.3, and 3.0 mg/kg LY, respectively. Therefore, LY prolonged VERP and APDE and affected DFT in the same saturating dose-response manner. LY may facilitate defibrillation by increasing the duration of postshock refractoriness. PMID- 8720421 TI - Comparative effects of idazoxan, prazosin, and yohimbine on coronary ligation induced arrhythmias in spontaneously hypertensive rats. AB - We investigated whether certain drugs with alpha-adrenergic antagonist activity display anti-arrhythmic effects in hypertensive animals subjected to acute coronary artery ligation. The left anterior descending coronary artery (LAD) was ligated in open-chest pentobarbital-anesthetized spontaneously hypertensive rats (SHR); arrhythmias were subsequently recorded for 30 min. Drugs were administered intravenously, (i.v.) 5 min before ligation. The effects of yohimbine and idazoxan were compared with those of prazosin. Prazosin (100 mu g/kg) increased the occurrence of ventricular tachycardia (VT). In contrast, yohimbine 1.6 mg/kg decreased both the occurrence and the duration of VT and the occurrence and the duration of ventricular fibrillation, (VF). The results obtained with idazoxan 1 mg/kg were similar to those with yohimbine. The ECG alterations induced by coronary artery ligation in rats treated with yohimbine and idazoxan were more pronounced than in controls and in rats treated with prazosin, suggesting that the antiarrhythmic effects observed were not mediated by antiischemic activity. The protective effects against ligation-induced arrhythmias were preceded by a hypotensive effect and a decrease in the rate-pressure product in yohimbine treated but not in idazoxan-treated animals. In rats treated with prazosin, more arrhythmic events were observed, although hemodynamics were similar to those in rats treated with yohimbine. Our results suggest that the yohimbine-induced antiarrhythmic action is not due to an alteration of conduction or repolarization rates. In this model, yohimbine and idazoxan appear to protect against ligation induced arrhythmias. These data suggest that drugs with alpha-adrenergic properties might influence the nervous drive to the heart in SHR with cardiac ischemia. However, further investigations are needed to ascertain whether the alpha-adrenoceptor blockade participates in this effect. PMID- 8720422 TI - Effects of manipulation of dietary cholesterol on the function of the thoracic aorta from New Zealand white rabbits. AB - Animal studies, while generally showing loss of endothelium-dependent responses after an elevation in plasma cholesterol, have provided conflicting reports with regard to recovery of function after normalisation of cholesterol level. Therefore, we assessed changes in vascular function after a period of hypercholesterolaemia and the subsequent effect of normalisation of cholesterol levels. Contractile responses to phenylephrine (PE) and endothelium-dependent relaxation in response to carbachol were examined in thoracic aorta from New Zealand White rabbits (NZW) fed a 0.3% cholesterol diet for 20 weeks, from NZW fed a 0.3% cholesterol diet for 20 weeks, followed by standard diet for 20 more weeks, and from their respective age-matched controls. Cholesterol levels were increased in rabbits receiving the 0.3% cholesterol diet (12.7 +/- 3.2 mM; 0.5 +/ 0.1 mM control) and returned to normal when standard diet was reintroduced (0.8 +/- 2.0 mM). Contractile responses were not affected by the period of hypercholesterolaemia. Carbachol-induced relaxation of a submaximal PE contraction was impaired after the period of hypercholesterolaemia (Emax 69 +/- 9%; 95 +/- 3% age-matched (control); the effect was reversed after reintroduction of standard diet (Emax 79 +/- 6%; 82 +/- 2% age-matched control). Our results demonstrate that endothelium-dependent relaxation is impaired after a long-term 0.3% cholesterol diet. Furthermore, after reintroduction of a normal diet, there is no further impairment of endothelium-dependent relaxation and endothelium function improves. PMID- 8720423 TI - Chronic nifedipine treatment diminishes cardiac inotropic response to nifedifine: functional upregulation of dihydropyridine receptors. AB - Chronic treatment with nifedipine induces up-regulation of functional active Ca2+ channels in cardiac muscle membranes. Adult male New Zealand White rabbits (NZW) were treated with nifedipine (20 mg/day) for 25 days. In isovolumic perfused hearts at constant coronary flow and heart rate (HR) the left ventricular developed pressure (LVDP) and its first derivative (dP/dt) were monitored. Basal contractility and contractility at different end-diastolic volumes (EDV) were higher in nifedipine-treated animals, with no changes in diastolic chamber stiffness. Dose response to nifedipine in pretreated animals showed less decrease in contractility than in controls [ED50 = 1.09 +/- 0.09 x 10-7 (control) and 1.55 +/- 0.17 x 10-7 M nifedipine (treated) (p < 0.05)]. Ca2+ channel density was assessed by specific binding at the dihydropyridine receptor with [methyl-3H]PN 200-110. In cardiac membranes, maximal binding capacity (Bmax) was 269 +/- 38 (n = 7, control) and 429 +/- 46 fmol/mg protein (n = 7, treated) (p < 0.05), without significant changes in dissociation constant. In addition, we noted no changes in dihydropyridine (DHP) binding sites in aortic membranes. Our results offer a possible explanation for the lack of decrease in contractility despite the persistent hypotensive effect in hypertensive patients during chronic treatment with nifedipine. PMID- 8720424 TI - Inhibition of nitric oxide synthesis reduces coronary blood flow response but does not increase cardiac contractile response to beta-adrenergic stimulation in normal dogs. AB - Whether the nitric oxide (NO) system constitutively present in the normal myocardium and resistance coronary vessels regulates basal cardiac contractility and coronary blood flow (CBF), as well as their responses to beta-adrenergic stimulation in intact heart, remains controversial. We examined the effects of low and high doses of NG-nitro-L-arginine methyl ester (L-NAME) (10 and 100 mu g/kg/min for 10 min), an NO synthase inhibitor, as well as D-enantiomer administered into left circumflex (LCX) artery on responses of left ventricular (LV) dP/dt, regional wall thickening in LCX region and LCX blood flow to graded intracoronary doses of isoproterenol (ISO 0.002-0.016 mu g/kg/min) in normal dogs. Intracoronary L-NAME, which was associated with dose-related reductions in acetylcholine (ACh)-induced coronary vasodilation, significantly reduced baseline LCX blood flow and its response to ISO. However, L-NAME did not change baseline LV contractility as assessed by LV dP/dt and regional wall thickening, nor did it increase its response to ISO. D-Enantiomer was ineffective in reducing baseline LCX blood flow as well as its response to ISO. These results indicate that constitutive NO formation in the vasculature contributes to basal coronary vascular tone as well as resistance adjustments during beta-adrenergic stimulation. However, NO formation in the normal myocardium did not influence basal cardiac contractility; nor did it increase cardiac response to beta adrenergic stimulation. PMID- 8720425 TI - Twenty-four-hour antihypertensive efficacy of felodipine 10 mg extended-release: the Italian inter-university study. AB - We assessed the 24-h antihypertensive efficacy of an extended-release (ER) 10-mg formulation of the dihydropyridine felodipine in mild-to-moderate essential hypertension [World Health Organization (WHO) stage I-II]. Thirty patients, 23 men and 7 women, aged 37-70 years (mean 53 +/- 9 years) participated in a double blind, randomized, cross-over study of felodipine 10 mg ER versus placebo. An ambulatory daytime diastolic blood pressure (DBP) >90 mm Hg at the end of a 4 week run-in period was necessary to enter the 10-week treatment phase. Twenty nine patients completed the treatment phase. Twenty-two underwent a 2-day single blind placebo follow-up to assess residual drug effects. All patients underwent ambulatory BP monitoring (ABPM) by Spacelabs 90207 recorders. Recorders were programmed to make automatic BP and heart rate (HR) measurements every 15 min throughout the 24 h. Felodipine 10 mg ER significantly (p < 0.01) reduced ambulatory systolic BP (SBP) and DBP values throughout the 24-h, day (7 a.m. to 11 p.m.) and night (11 p.m. to 7 a.m.) periods, but not influencing average ambulatory HR values. Trough-to-peak (T/P) ratios, calculated on the average ambulatory BP values measured in the 7-9 a.m. 2-h interval of the second day of ABPM (before the new drug administration: trough) and in the 10 a.m. to 12 noon 2 h interval of the first day of ABPM (peak BP-lowering effect), were 0.71 and 0.58 for SBP and DBP, respectively. Individual T/P calculations, after post hoc selection of nonresponders, gave superimposable results, the consistency of which was judged on mean, median, and confidence intervals (CI). However, the wide variability of the individual T/P ratios suggests that this method cannot be the only means to evaluate the duration of action of an antihypertensive drug by ABPM. The long-acting BP-lowering drug effect was clearly shown by the ABPM performed in the follow-up when SBP and DBP average values of the 24-h, day, and night periods were still reduced. Felodipine 10 mg ER effectively reduced BP in patients with mild-to-moderate hypertension, showing prolonged duration of its antihypertensive action beyond the time of the next dose. PMID- 8720426 TI - Biphasic antagonisms by beta-blockers against positive inotropic response through beta1-adrenoceptors in isolated canine right ventricular muscles: possible involvement of two beta 1-adrenoceptor subtypes. AB - In isolated canine right ventricular muscles, we investigated the differences in antagonisms by beta-blockers against the positive inotropic effects (PIEs) of isoproterenol, a nonselective agonist, and T-0509, a beta1-selective agonist. The selective beta1-blockers atenolol and bisoprolol antagonized the PIE of T-0509 monophasically in Schild analysis, showing pA2 values of 7.05 and 7.63, respectively. On the other hand, both blockers produced biphasic antagonism against the PIE of isoproterenol (ISO); therefore, two pKB values were obtained (7.75 and 4.25 and 7.82 and 5.76, respectively). Nadolol, a nonselective beta blocker, also antagonized the PIE of T-0509 monophasically (pA2 value 7.58), but antagonized the PIE of ISO biphasically (pKB values 7.42 and 4.39). Because the different mode of antagonism by three beta-blockers between T-0509 and ISO could not be explained by the selectivities of beta-agonists and blockers for beta1- and beta2-adrenoceptors in the heart, two subtypes of beta1-adrenoceptors may exist together in canine ventricular muscles, and atenolol, bisoprolol, and nadolol may act as antagonists for the two subtypes with two different affinities. PMID- 8720427 TI - Kinetics, safety, and efficacy of ramipril after long-term administration in hemodialyzed patients. AB - We studied the efficacy and safety of ramipril and the kinetics of its active moiety ramiprilat in 12 hypertensive patients receiving regular hemodialysis, after a single dose and after long-term (28 days) administration. Patients received 2.5 mg ramipril after each hemodialysis. On days 1 and 29, ramipril was administered 4 h before the hemodialysis and serial blood samples were obtained for 9 h for determination of pharmacokinetic parameters. Tolerability was good, and all patients completed the study. There was a high degree of angiotensin converting enzyme (ACE) inhibition throughout the study. Ramipril had a clear-cut antihypertensive effect. Long-term administration of ramipril did not modify the time to peak ramiprilat concentration, but increased the mean maximal concentration significantly: 20.2 +/- 12.7 vs. 10.4 +/- 7.1 ng center dot ml-1. The mean accumulation ratio was 2.2. Ramiprilat hemodialysis clearance was 31.7 ml/min (range 4.2-64.9 ml/min) on day 1 and 21.0 ml/min (range 7.9-56.5 ml/min) on day 29. Ramipril 2.5 mg, administered after hemodialysis, appears to be safe and effective in hypertensive patients receiving periodic hemodialysis. Despite an increase in ramiprilat concentration from day 1 to day 29, the steady state was reached. We describe the role of nonrenal clearance of ramiprilat. PMID- 8720428 TI - Effect of long-term treatment with selective vasopressin V1 and V2 receptor antagonist on the development of heart failure in rats. AB - Vasopressin has been implicated in the pathogenesis of heart failure as one of the most potent vasoconstrictors. However, whether the increase in plasma vasopressin levels modifies the pathophysiology of heart failure remains unknown. To investigate the effect of long-term inhibition of vasopressin in the development of heart failure, we administered a selective, orally effective, nonpeptide vasopressin antagonist, the V1 receptor antagonist OPC-21268 (100 mg center dot kg-1 center dot day-1) or a V2 receptor antagonist, OPC-31260 (20 mg center dot kg-1 center dot day-1) to rats with heart failure induced by the creation of an aortocaval fistula (AVF) and to sham-operated rats for 4 weeks, beginning on the first postoperative day. The heart failure in this experiment was characterized by an increase in the weights of the right and left ventricles, the lungs, and the right and left appendage, increase in left ventricular end diastolic pressure (LVEDP), increase in right ventricular systolic pressure (RVSP), increase in right atrial pressure (RAP), and an increase in the plasma level of atrial natriuretic peptide (ANP) as compared with no change in sham operated rats. There were no differences in shunt ratio between treated and untreated heart failure groups. Chronic administration of the V2 receptor antagonist OPC-31260 significantly reduced the weight of the right ventricle (1.17 +/- 0.39 vs. 0.90 +/- 0.13 g/kg, p < 0.05), RVSP (53 +/- 18 vs. 39 +/- 4 mm Hg, p < 0.05), LVEDP (11.8 +/- 5.2 vs. 6.5 +/- 2.8 mm Hg, p < 0.05) and the plasma concentrations of ANP (554 +/- 271 vs. 193 +/- 39 pg/ml, p < 0.05) as compared with the values of rats with untreated HF. Chronic treatment with the V1 receptor antagonist OPC-21268 did not alter hemodynamics, organ weights, or hormone concentrations. These results suggest that vasopressin did not contribute mainly to the maintenance of systemic hemodynamics through the V1 receptor in this heart failure model. Vasopressin may play a role, at least in part, in the fluid retention in the development of heart failure through the V2 receptor. OPC 31260 may present a new approach to the treatment of heart failure. PMID- 8720429 TI - Recombinant lys-plasminogen given before, but not after, recombinant tissue-type plasminogen activator markedly improves coronary thrombolysis in dogs: relationship of thrombolytic efficacy with parameters of fibrinolysis. AB - Recombinant tissue-type plasminogen activator (rt-PA) administration rapidly restores blood flow in thrombosed coronary arteries, but coronary arteries often reocclude after initial thrombolysis. This occurs because of the short half-life of rt-PA and rapid increase in plasminogen activator inhibitor (PAI-1) and alpha2 antiplasmin levels in plasma. We hypothesized that administration of lys plasminogen, which binds to fibrin with 10 times greater affinity and results in a loose fibrin structure (as compared with native glu-plasminogen), before rt-PA would enhance the thrombolytic efficacy of rt-PA and modulate parameters of fibrinolysis. To examine this hypothesis, dogs with electrically induced stable thrombus in the left anterior descending coronary artery (LAD) were treated with saline (group A, n = 9) or lys-plasminogen (group B, 2 mg/kg, n = 5), followed 10 min later by rt-PA (1 mg/kg in 20 min). Four other dogs with occlusive LAD thrombus were first given rt-PA, followed by lys-plasminogen (2 mg/kg) 50 min later (group C). Lys-plasminogen given before rt-PA restored flow in all dogs in 14 +/- 4 min (vs. 22 +/- 9 min in group A, p < 0.05), continuing > 2 h (vs. 41 +/ 15 min in group A, p < 0.02). Lys-plasminogen given after rt-PA did not potentiate the effect of rt-PA. Plasma t-PA antigen concentrations were highest in group B dogs at 2 h after rt-PA infusion. PAI-1 and alpha2-antiplasmin plasma levels were suppressed in all dogs receiving lys-plasminogen whether it was given before or after rt-PA. Therefore, lys-plasminogen given before rt-PA markedly potentiates the effect of rt-PA and alters the parameters of fibrinolysis. In contrast, lys-plasminogen given after rt-PA does not influence the thrombolytic effect of rt-PA, whereas it suppresses PAI-1 and alpha2-antiplasmin levels in plasma. This study also suggests that binding of plasminogen to the clot is more important than the plasma levels of PAI-1 and alpha2-antiplasmin. PMID- 8720430 TI - Interaction of lacidipine with its receptor binding site supports a slow onset and long duration of action. AB - [3H]lacidipine binding to its receptor was characterized to explain its slow onset and long duration of antihypertensive activity. Binding parameters were studied in guinea pig myocardial and cerebral membrane preparations and compared with another dihydropyridine (DHP) calcium antagonist, isradipine. Lacidipine binds competitively to the DHP calcium antagonist receptor of the L-type calcium channel. The binding is allosterically modulated by verapamil and D-cis diltiazem and activated/inhibited by divalent cations. Association and dissociation kinetics of the binding of lacidipine to the receptor were significantly slower than those of isradipine. In addition, the Bmax of lacidipine binding in guinea pig heart microsomes was significantly higher than those of other dihydropyridine calcium antagonist. The results indicate that the slow onset and long duration of action of lacidipine can be explained principally on the basis of the binding characteristics. Although no biphasic receptor binding kinetics could be detected, a fast equilibrium between the receptor and a second compartment, due to the high lipophilicity of lacidipine, cannot be excluded. PMID- 8720431 TI - Endothelial Gi protein expression is markedly low in human coronary microvessels. AB - Gi protein functionally mediates endothelium-dependent relaxations in large epicardial coronary arteries but not in small coronary arteries, which suggests a different involvement of Gi protein in the endothelium-dependent relaxations between large and small coronary arteries. We previously showed that endothelial Gi protein is present in human epicardial coronary artery. In the present study, we examined the expression of endothelial Gi protein in human coronary microvessels. Immunohistochemical staining with a specific antibody against human Gi protein was performed in intramyocardial coronary microvessels and vasa vasorum from 34 autopsy cases. The immunoreactive levels of the endothelial Gi protein were semiquantitated into four grades (none, 0; slight, +1; moderate, +2; high, +3), and the mean value of the ratings of all endothelial cells was then used as an index of the endothelial Gi protein expression of the vessel. The immunoreactive levels of the endothelial Gi protein were extremely low in intramyocardial coronary microvessels and in vasa vasorum, irrespective of the age of the patients, the presence or absence of coronary risk factors, or the influence of medical treatments. These results may therefore explain in part why endothelium-dependent relaxations in coronary microvessels are not functionally mediated by Gi protein. PMID- 8720432 TI - Effect of atrial natriuretic factor on skin microcirculation versus skeletal muscle blood flow. AB - The response of the skin microcirculation and of forearm skeletal muscle blood flow to infusion of alpha-human (99-126) atrial natriuretic factor (ANF) into the brachial artery was investigated in 15 young (18-25 years) healthy volunteers in a double-blind, randomized, placebo-controlled study. The forearm blood flow (FBF) was measured with venous occlusion plethysmography, and the skin flux was measured by using laser Doppler fluxmetry (LDF). Dose-response curves were made using increasing dosages of ANF: 1, 10, and 100 ng/min/dl forearm volume. The FBF showed a significant, dose-dependent increase during ANF infusion, averaging 107 + 22% during the highest ANF dosage, as compared with -5 +/- 9% during placebo (p < 0.001). For the LDF, these numbers were 34 +/- 21 and -6 +/- 10%, respectively (NS). In two subgroups of subjects, the effect of ANF on microvascular reactivity was assessed by registering the vasoconstrictor response to cold exposure (n = 7) and the vasodilator response to arterial occlusion (n = 7). ANF did not change the microvascular response to these stimuli. ANF induces a dose-dependent increase in skeletal muscle BF without a relevant response in the skin microcirculation. ANF does not play an important role in the regulation of skin perfusion. PMID- 8720433 TI - Morphological and electrophysiological properties of centrifuged stratified Xenopus oocytes. AB - To separate and concentrate various cytoplasmic organelles in wild type and albino Xenopus oocytes, defolliculated cells were loaded on a Ficoll-400 gradient and centrifuged. Optimum results were obtained with centrifugations at 10,000 g for 5 min at 20 degrees C. The cells became pear-shaped and appeared stratified with the white lipid yolk on top, an intermediate transparent zone of about 100 300 microns, and the greenish protein yolk at the bottom. To determine the cellular constituents, particularly of the transparent zone, electron microscopy was performed. The transparent zone was found to contain (from animal to vegetal) the various endoplasmic reticula, a layer of mitochondria, cytoplasm enriched in ribosomes and the depressed nucleus. In centrifuged stratified wild type oocytes, most of the pigment was layered on top of the protein yolk. The typical cortical aspects of the oocyte persisted. Centrifuged albino oocytes had a very pronounced transparent zone with sharp transitions to the lipid phase and to the protein yolk. The resting membrane potentials of centrifuged oocytes were between -35 and -65 mV, and the membrane resistances were in the 500 k omega to 1 M omega range. Under voltage clamp conditions, the oocytes exhibited Ca(2+)-activated Cl- currents with biphasic kinetics and spontaneous oscillations of these currents. It is concluded that centrifuged stratified oocytes have normal electrophysiological properties, and that they are a suitable preparation to study the contribution of various cellular organelles to the propagation of second messengers in the cytosol. PMID- 8720434 TI - Immunocytochemical localization of actin in the nucleolus of rat oocytes. AB - In order to determine the localization of actin, growing and fully grown rat oocytes were immunocytochemically examined using a post-embedding ultrastructural protein-A gold technique. In quiescent oocytes, the nucleoplasm showed slightly lower levels of actin signal when compared to the surrounding cytoplasm. The highest levels of labeling were found on nucleoli showing a reticular type morphology. In oocytes at the diakinesis stage in which nucleolar compaction had occurred, the levels of labeling increased by 5-6 times those found in quiescent oocytes. Except for conspicuous accumulation of actin under the plasma membrane, compact nucleoli had significantly higher levels of labeling when compared with those found on the general cytoplasm, while the nucleoplasm with homogeneously dispersed chromatin showed significantly lower levels of associated actin signal than the general cytoplasm. In oocytes at metaphase I, the cytoplasmic region had comparable or lower levels of labeling than the cytoplasm of oocytes at diakinesis. The meiotic spindle embedded in material with medium electron density showed a similar level of labeling as the surrounding cytoplasm. On the other hand, significantly higher levels of associated actin were observed on the chromosomes of metaphase I. The actin signals were dispersed over the chromosomes and not concentrated on a specific region. These results suggest that nuclear actin may be involved in the process of chromosome construction and also the formation of the compacted structure of the nucleolus. PMID- 8720435 TI - Flow cytometric investigation of neutrophil activation pathways by n-formyl-Met Leu-Phe and phorbol myristate acetate. AB - Recent evidence suggests that multiple pathways exist in PMN activation and that specific leukocyte response may be due to the activation of a particular signaling pathway. Using flow cytometry, PMN activation pathways were studied through the parallel comparison of n-formyl-Met-Leu-Phe (fMLP)- and phorbol-12 myristate-13-acetate (PMA)-induced stimulation and by simultaneous assays for CD11b expression and morphology. The maximal CD11b expression was higher with PMA than with fMLP, suggesting different activation pathways. Under these experimental conditions, a morphological response to fMLP was not observed. However, significant shape change was detected in PMA treated samples and was suppressed by either the removal of extracellular calcium or staurosporine at the concentrations above 14.5 microM. Calcium ionophore induced a similar light scattering pattern to that by PMA and enhanced CD11b expression, both of which were not inhibitable by staurosporine. These observations, for the first time, indicated that Ca2+ was a mediator in activation processes and that the treatment of PMN with PMA resulted in Ca2+ influx. PMID- 8720436 TI - In isolated human centrosomes, the associated kinases phosphorylate a specific subset of centrosomal proteins. AB - Several studies have shown that kinases and phosphatases can interact with the centrosome during interphase and mitosis suggesting that centrosomal components might be the targets of these enzymes. The association of the cAMP-dependent protein kinase type II and the mitotic kinase p34cdc2 with centrosomes from human lymphoblast cells has previously been shown (Keryer et al, 1993, Exp Cell Res 204, 230-240; Bailly et al, 1989, EMBO J 8, 3985-3995). In this paper we demonstrate that isolated centrosomes are able to phosphorylate a few number of centrosomal proteins (M(r) 230-220000; 135000 and 50000) and also H1 histone. The phosphorylation of H1-histone is cell cycle dependent and modulated by phosphatases. The use of kinase and phosphatase inhibitors and the addition of the catalytic subunit of cAMP-dependent kinase or of cyclinB-p34cdc2 kinase showed that both kinases phosphorylate the same centrosomal substrates. In addition two centrosomal proteins (M(r) 100000 and 37000) were phosphorylated only by p34cdc2 kinase. Although the low amount of centrosomal proteins precluded a full characterization of these substrates we discuss the identity of the major centrosomal phosphoproteins by comparison with proteins known to associate with microtubule-organizing centres or mitotic spindles. Our results raise also the intriguing possibility that the cAMP-dependent protein kinase could be regulated by the mitotic kinase at the entry of mitosis. PMID- 8720437 TI - Cell-cycle dependent biosynthesis and localization of p53 protein in untransformed human cells. AB - Localization of p53 in human cultured lymphocytes and in cultured skin fibroblasts was studied by immuno-fluorescent microscopy and post-embedded immunoelectron microscopy using Lowicryl K4M. In quiescent lymphocytes, p53 was found in small amounts in both the cytoplasm and the nucleus. p53 in the nucleus was found associated with the non-chromatin structure. At 24 h or 72 h of PHA stimulation, p53 increased markedly just beneath the plasma membrane and in the nucleus, which stained diffusely with anti-p53. In resting fibroblasts, small amounts of p53 were present in both the cytoplasm and the nucleus. After 16 h of stimulation of confluent-resting fibroblasts by trypsinization and replating, a phase just prior to the initiation of DNA synthesis, p53 slightly increased in both the cytoplasm and the nucleus. Afterwards, p53 was present predominantly in the cytoplasm, closely associated with the cytoskeletal actin filaments. In mitotic cells, p53 was distributed throughout the cytoplasm. When fibroblasts were extracted with saponin, p53 was still associated with the actin filaments, as well as mitochondrial membranes and granular structures of the nuclear matrix. Our data suggest that the initial increase of p53 in cells that enter the cell cycle through G1 first bind to the actin cytoskeleton, and that some of the p53 then move into the nucleus to initiate gene activation and DNA synthesis for cell proliferation. This implies that there is some functionally significant interaction between p53 and actin in the cells. PMID- 8720438 TI - Cell-mediated cytotoxicity can be regulated by p53 tumor suppressor gene activity in vitro. AB - The wild-type human p53 tumor suppressor gene was tested for its ability to modulate cytotoxic activity of in vitro activated peripheral blood lymphocytes. Peripheral blood mononuclear cells (PBMCs) were stimulated by phytohemagglutinin (PHA), interferon alpha 2b (IFN alpha 2b), interleukin 2 (IL-2) or their combinations to induce cytotoxicity. This stimulation significantly increased the percentage of cells expressing p53, which was at its maximum when induced by IL-2 combined with IFN alpha 2b. The role of p53 in the modulation of different aspects of cytotoxic activity of these cells was analyzed by studying the effects of p53 abrogation by antisense oligonucleotide (p53 AS) treatment in comparison with p53 sense or scrambled (missense) oligonucleotide (p53 S or p53 MS) treatment. We show that p53 plays a key role through induction of apoptosis in target cells (tumor necrosis factor pathway) rather than through osmolytic degeneration (perforin pathway) which is only slightly increased by p53 abrogation. Meanwhile, in vitro abrogation of p53 expression in PBL was found to be accompanied by an increase of CD8+ lymphocytes and an important increase of the CD56 'bright' NK cell sub-population. PMID- 8720439 TI - Normal human endometrial cells in culture: characterization and immortalization of epithelial and stromal cells by SV 40 large T antigen. AB - Thirty endometrial biopsies were cultured in order to separate stromal and epithelial cells. Using epidermal growth factor (EGF), cortisol, cholera toxin, insulin with 5% horse serum for epithelial cells or a medium with 20% fetal calf serum for stromal cells, we could specifically enrich endometrial culture in epithelial or stromal cells and culture them for 1 or 2 months. These cultures retained the phenotypic characteristics of epithelial (cytokeratins, mucin HMFG 1) and stromal (vimentin, smooth muscle actin, desmin) lineage by immunostaining analysis. Epithelial and stromal cultures from one individual were respectively immortalized by the SV 40 large T antigen. The immortalized cell lines kept the phenotype of the normal cells from which they derived. PMID- 8720440 TI - Multidrug-resistance (MDR) phenotype of human osteosarcoma cells evaluated by quantitative morphological and electron microscopy analyses. AB - Multidrug-resistant (MDR) variants of a human osteosarcoma cell line (U-2 OS) have been recently obtained by continuous exposure to doxorubicin (DX). The growth and phenotypic characteristics of these cell lines have been demonstrated to be related to the level of expression of P-glycoprotein. In this work, the morphological changes associated with MDR have been evaluated by quantitative image analysis and transmission electron microscopy. Resistant cells present morphological changes with respect to sensitive cells at both cytoplasmic and nuclear level. Some of these changes appear to be related to the degree of resistance but not to the direct presence of DX, since deprived cells maintain some modified characters, while others are partly lost. These findings suggest that DX exposure affects cell metabolism causing progressive changes of the cell morphotype. PMID- 8720441 TI - Production of transforming growth factor (TGF) beta by fetal lung cells. AB - TGF beta is supposed to play an important role in the process of epithelial maturation in the developing fetal lung. Using an immunofluorescence approach, we showed that fetal rat lung fibroblasts elaborate the three TGF beta isoforms known in mammals (TGF beta 1, beta 2 and beta 3) whereas epithelial cells appear to synthesize only TGF beta 1 and beta 3. Isolated fibroblasts secrete the three isoforms. Biological assay of TGF beta activity in fibroblast-conditioned media did not reveal significant changes according to the stage when fibroblasts were isolated. PMID- 8720442 TI - Experimental vasectomy and testicular structure. AB - We have performed an experimental study on rats and dogs to evaluate the long term effects (from 1 to 12 months) of vasectomy on the structure of the testis. From four months after vasectomy onwards, the specimens showed very important changes in the seminiferous epithelium and Sertoli cells, with an obvious thickening of the basement membrane that supports the epithelium. The deterioration depended on the time passed and, over six months after vasectomy, the alterations were very clear and the seminiferous tubules became atrophic and shrunk, sometimes without any remains of seminiferous epithelium and with an important hypertrophy of the interlobular interstitial tissue, although we did not see an increase in the number of Leydig cells. Alterations due to vasectomy depend on the animal species, the peculiarities of techniques and, of course, the time passed after surgery. PMID- 8720443 TI - Tobacco smoke and age as risk factors in emphysema. Morphometrical study on the rat. AB - During ageing, a progressive deterioration in the pulmonary function, which can be accelerated by exposure to tobacco smoke, takes place. The hypothesis that the initial age of exposure to tobacco smoke is a factor of utmost importance in the development of emphysema is proposed. Eighty-six rats, aged nineteen months at the time of sacrifice, were used and were ordered into three groups: the first group consisted of unmanipulated animals; the second, of animals which had been exposed to tobacco smoke from the age of twelve months to the age of nineteen months; and the third, of animals which had been exposed to tobacco smoke from the age of nine months to the age of twelve months. The lungs of the animals were histologically processed for light microscopy and were studied morphometrically by computer. Eleven quantitative variables were quantified and ordered into three groups: variables related with alveolar enlargement; variables related with tissue loss; and variables related with the elastic fibre. The number of animals in which alveolar enlargement and tissue destruction concurred was counted, thus enabling the attributable and relative risks of developing emphysema to be calculated in the two groups of manipulated animals. From the results it is clear that, when compared with the unmanipulated group, the two groups which had been exposed to tobacco smoke displayed an increase in the variables which quantified alveolar enlargement and a decrease in those which measured tissue loss; these results were more significant in the third group (p < 0.001) than in the second (p < 0.05); significant differences were also found between these two groups of animals. The relative risk and attributable risks of developing emphysema were 2.41 and 28.15 respectively in the second group and 3.48 and 34.48 in the third group. Our results lead us to propose that the risk of developing emphysema exists in inverse proportion to the initial age of exposure to tobacco smoke. PMID- 8720444 TI - Ultrastructural study of the vascular response in small early gastric cancer. AB - The microvasculature of the stroma of four cases of small early gastric cancer (EGC) was investigated by conventional electron microscopy. Severe damage to small and large fenestrated capillaries was observed around endothelium-adherent, partially degranulated neutrophils. The findings suggest the existence of neutrophil-mediated injury of endothelial cells during the development of inflammatory responses in small EGC. The severely injured microvessels exhibited increase in vasopermeability, microhaemorrhage, and platelet aggregates. Other microvascular changes included endothelial cell and pericyte activation as well as basal lamina replications, indicative of repeated episodes of endothelial injury, necrosis and regeneration. This new capillary growth within the old basal laminas shared morphological features with a peculiar angiogenic process described in man and animals. PMID- 8720445 TI - Ultrastructure of liver from piglets fed Tower rapeseed oil. AB - Effect of Tower rapeseed oil (RO) containing lower (0.4%) erucic acid on the piglet liver was studied by electron microscopy. The animals were placed in two groups of four and were fed the diets for eight weeks. Animals in the treated group were given a basal diet comprising corn-soybean plus Tower RO at a 10% concentration; animals that were given the basal diet with no added oil served as the controls. Architecture of the liver from animals of the control group appeared normal. Ultrastructural changes in the liver of RO-fed animals included dilation of lumens and loss of microvilli in bile canaliculi. In addition, closely aligned mitochondrial cristae, elevated number of peroxisomes, and an unusual increase in the size and number of membrane bound spaces in zone 3 of liver acini were noticed. These alterations may reflect a disturbance in piglet liver functions following Tower RO feeding. PMID- 8720446 TI - Argyrophilic nucleolar organizer region (AgNOR) counting in astrocytic gliomas: prognostic value. AB - In 87 astrocytic gliomas the number of AgNORs/nucleus was retrospectively studied and data correlated with the histological type of the tumors and survival. All patients were treated by the same surgical team and with uniform criteria. Statistically significant differences (p < 0.01) were found in relation with the AgNOR averages among the histological types of tumors. A statistically significant linear correlation (p < 0.05) between the AgNOR values and survival of the patients was also found. Patients with mean AgNOR values higher than 2.23 and lower than 2.9 survived an average of 11.5 +/- 9.1 months vs. a survival in average of 24.4 +/- 34.1 months with mean AgNOR values under 2.23 (p < 0.05). Patients with AgNOR values higher than 2.9 survived, on average, 7.7 +/- 3.9 months. AgNOR counting in astrocytic gliomas is a reproducible, easy, quick method with prognostic value. AgNORs may be successfully applied in routine material to assess the growth potential of astrocytic gliomas. PMID- 8720447 TI - Fine structure of the retina and pigment epithelium in the creek chub, Semotilus atromaculatus (Cyprinidae, Teleostei). AB - The structure of the light- and dark-adapted retina, the pigment epithelium and the choroid of the creek chub, Semotilus atromaculatus (Cyprinidae, Teleostei) is examined by light and electron microscopy. An extensive network of vitreal blood vessels emanating from the hyaloid artery enters the eye with the optic nerve and overlies the inner limiting membrane. This membrane closely apposes the fine protrusions of the Muller cell processes which traverse the entire retina, dividing the inner retina into alternating fascicles of ganglion cells and optic axons. The inner nuclear layer consists of bipolar, amacrine, Muller cell soma and two layers of horizontal cells. The outer plexiform layer possesses both rod spherules and cone pedicles. Each rod spherule consists of a single synaptic ribbon in either a triad or quadrad junctional arrangement within the invaginating terminal endings of the bipolar and horizontal cell processes. In contrast, cone pedicles possess multiple synaptic ribbons within their junctional complexes and, in the light-adapted state, the horizontal cell processes show spinule formation. Four photoreceptor types are identified on morphological criteria; unequal double cones, large single cones, small single cones and rods. All but the small single cones are capable of retinomotor responses. The rod to cone ratio is approximately 5:1 and the rods form two ill-defined rows in the light-adapted condition. The retinal pigment epithelium possesses two types of osmiophilic granules. These are bound within slender microvilli and migrate vitread to surround the photoreceptors in response to light. Bruch's membrane is trilaminar and the vascularised choroid consists of up to three layers of melanocytes. The endothelial borders of the choroidal blood vessels abutting the outer lamina of Bruch's membrane are fenestrated. PMID- 8720448 TI - Ultrastructure and organisation of the retina and pigment epithelium in the cutlips minnow, Exoglossum maxillingua (Cyprinidae, Teleostei). AB - The structure of the light- and dark-adapted retina, pigment epithelium and choriocapillaris of the cutlips minnow, Exoglossum maxillingua (Cyprinidae, Teleostei) is examined by light and electron microscopy. A pronounced vitreal vascularisation overlies the inner retina where the blood vessel walls, the inner limiting membrane and the Muller cell endfeet are all closely apposed. The thick Muller cell processes divide the inner plexiform layer and nerve fibre layer into discrete compartments. The ganglion cells do not form fascicles and lie within both the ganglion cell and inner plexiform layers. The inner nuclear layer consists of amacrine, bipolar, Muller cell somata and two rows of horizontal cells. The photoreceptor terminals comprise either multiple (3-5 in cone pedicles) or single (rod spherules) synaptic ribbons. These photoreceptor terminals form either a triad (rods and cones) or a quadrad (cones) arrangement of contact with the invaginating processes of the inner nuclear layer cells. The horizontal cell processes of the cone photoreceptor terminals reveal spinule formation in the light-adapted condition. Five photoreceptor types are classified using morphological criteria; triple cones, unequal double cones, large single cones, small single cones and rods. The ratio of rods to cones is approximately 7:1. All photoreceptor types show retinomotor responses. Only the cones possess accessory outer segments but both rods (8-11) and cones (15-19) possess calycal processes. The retinal pigment epithelium displays retinomotor responses where pigment granules within fine apical processes move vitread to mask the rods in the light. The cells of the retinal pigment epithelium are joined by various types of junctions and contain numerous phagosomes, mitochondria and polysomes. Bruch's membrane or the complexus basalis is trilaminate with two types of collagen fibrils comprising the central layer. The endothelia of the blood vessels of the choriocapillaris, facing Bruch's membrane, are fenestrated. Two to three layers of melanocytes interspersed between large thin-walled capillaries and several layers of collagen fibrils comprise the choriocapillaris. PMID- 8720449 TI - Fine structure of the retinal pigment epithelium of the barred owl (Strix varia). AB - The retinal pigment epithelium (RPE) as well as the choriocapillaris and Bruch's membrane (complexus basalis) have been studied by light and electron microscopy in the barred owl (Strix varia). The RPE consists of a single layer of cuboidal cells joined laterally by a series of tight junctions that forms part of the blood-ocular barrier. Basally (sclerally) the retinal epithelial cells display numerous deep infoldings while apically (vitreally) microvillar processes interdigitate with the photoreceptor outer segments. Internally the RPE cells show a large vesicular nucleus, plentiful smooth endoplasmic reticulum (SER) and polysomes but very little rough endoplasmic reticulum (RER). Numerous pleomorphic (including ring-shaped) mitochondria are basally located. In the light-adapted state the small melanosomes are almost exclusively located within the apical process indicating that retinomotor movements probably occur. Phagosomes and lysosome-like bodies are present as are myeloid bodies which may show ribosomes on their outer surface. Bruch's membrane is typical of avian species in that it is pentalaminate and the central lamina densa is displaced towards the choroid. The choriocapillaris endothelium is thin but only minimally fenestrated facing Bruch's membrane. Most fenestrations present show a single-layered diaphragm while others display a double-layered diaphragm as noted in other avian species. PMID- 8720450 TI - Fine structure of the retinal photoreceptors of the barred owl (Strix varia). AB - The photoreceptors of the barred owl (Strix varia) consist of rods, single cones and unequal double cones present in a ratio of about 35:1:3. In the light-adapted condition the rods are of uniform diameter along their entire length and are therefore not felt to undergo photomechanical changes. The rod outer segment consists of a stack of scalloped bimembranous discs enclosed in a limiting membrane. The rod inner segment displays an ellipsoid of mitochondria, much rough endoplasmic reticulum (RER), numerous polysomes, Golgi zones and autophagic vacuoles, but no hyperboloid of glycogen. Single cones show a slightly tapered outer segment and a heterogeneous oil droplet along with an ellipsoid of mitochondria at the apex of the inner segment. Double cones consist of a larger chief member which also displays a heterogeneous oil droplet and a slightly smaller accessory member which does not. Both members of the double cone as well as the single cones show plentiful polysomes and RER as well as Golgi zones in the inner segment, but none of the cones possessed a condensed paraboloid of glycogen. The contiguous membranes of the chief and accessory cones displayed a few presumed junctional complexes. Judging by their elongated shape in the light adapted state, cones in this species do not undergo retinomotor movements. Rods and both types of cones have both invaginated (ribbon) and superficial (conventional) synaptic sites. PMID- 8720451 TI - Fine structure of the pecten oculi of the barred owl (Strix varia). AB - The pecten oculi of the barred owl (Strix varia) has been examined by light and transmission electron microscopy. The pecten in this species is of the pleated type and is small in comparison to the size of the ocular globe. The pecten consists of 8-10 accordion-like folds that are linked apically by a pigmented tissue bridge. Each fold contains numerous capillaries, larger supply and drainage vessels, and abundant pleomorphic melanocytes. Most of these capillaries are extremely specialized vessels that possess plentiful microfolds on both the luminal and abluminal surfaces. Some capillaries however display only a few microfolds. The endothelial cell bodies are extremely attenuated, with most organelles located near the nucleus. All capillaries are surrounded by a very thick fibrillar basal lamina, which is thought to provide structural support to these small vessels. Pericytes are commonly found within these thickened basal laminae. Numerous melanocytes are also present, with processes that form an incomplete sheath around the capillaries. These processes are also presumed to provide structural support for the capillaries. As in other avian species, the morphology of the barred owl pecten is indicative of extensive involvement in substance transport. When compared to the pecten of more visually-oriented species, this pecten is smaller, has fewer folds, and displays a reduced number of microfolds within the capillaries. In these and other features, the barred owl pecten is similar to the pecten of the great horned owl (Bubo virginianus). PMID- 8720452 TI - Major histocompatibility complex expression in muscle of rats with graft-versus host disease. AB - Immunohistochemical examination of rat skeletal muscle during graft-versus-host disease (GVHD), a systemic immune reaction, was performed to investigate specific immune reactivities focusing on major histocompatibility complex (MHC) expression and inflammatory cell infiltration of skeletal muscle during a systemic immune reaction. MHC class II expression and inflammatory cell infiltration did not increase. MHC class I was expressed along the contour of muscle fibres, and most strongly expressed by the cells which were distributed throughout the endomysium and perimysium. Seventy-six percent of these MHC class I+ cells carried endothelial cell-markers, while 24% of them did not. The latter cells were revealed not to be inflammatory cells such as lymphocytes, granulocytes or macrophages when examined by immunostaining using several exudate-cell markers. Neither were they myosatellite cells because they were located outside the basement membrane. These results may be useful for considering animal models of inflammatory myopathies such as polymyositis and dermatomyositis. PMID- 8720453 TI - Fine structure of the acinar and duct cell components in the parotid and submandibular salivary glands of the rat: a TEM, SEM, and HRSEM study. AB - Parenchymal and stromal components of the rat parotid and submandibular glands were examined by conventional and high resolution scanning electron microscopy (HRSEM). Freeze-fractured specimens were subjected to HCl and NaOH extraction procedures to better differentiate connective tissue and cellular components. In addition, the internal three-dimensional morphology of the secretory acinar cells and duct cells was revealed by maceration with a dilute osmium tetroxide solution to selectively remove some of the cytoplasmic components. SEM and HRSEM examination of the HCl-treated samples of both glands revealed a fine filamentous network immediately surrounding each acinus. Coarser bundles of collagen that linked neighboring acini were also observed. NaOH-extracted samples selectively removed the cellular components and showed more clearly the three-dimensional structure of the connective-tissue stroma. A dense-collagenous network surrounded each lobule while more internal regions consisted of a honeycomb-like pattern of evacuated spaces previously occupied by secretory acini. These spaces were smoothened in appearance and often interconnected. Apically-located secretory granules and profiles of the rough endoplasmic reticulum and Golgi apparatus in perinuclear regions were encountered in the acinar and duct cells of macerated samples by HRSEM. In addition, a phenylephrine-induced experimental condition performed in some rats resulted in a significant increase in secretory granule size and density of the serous cells. PMID- 8720454 TI - Rhinophyma--unusual expression of simple-type keratins and S100A in sebocytes and abundance of VIP receptor-positive dermal cells. AB - Rhinophyma represents a severe variant of rosacea, a common mid-facial erythematous dermatosis. Increased blood flow and pooling in skin are thought to be involved in its pathogenesis. Since neuropeptides and their receptors are responsible for local blood flow regulation, immunolocalization for the vasoactive intestinal peptide (VIP)-receptor(R) was performed in slice biopsies taken from five patients with glandular rhinophyma. Additional immunostainings included intermediate filaments (keratin, vimentin) and neuroglandular antigen (NGA). In contrast to controls, rhinophyma disclosed not only a more dense distribution of VIP-R positive cells within the endothelium but immunoreactive perivascular large cells. The immature sebocytes stained positive with monoclonal antibody Cam5.2 against glandular antigens and polyclonal anti-S100A. Elastotic connective tissue in the dermis showed a strong immunoreactivity for vimentin and NGA. From these results we suggest that, (a) ligands of the VIP-R may contribute to vascular and dermal alterations in rosacea and (b) immature sebocytes show an unusual antigen expression of S100A and glandular keratin. PMID- 8720455 TI - Projection of forelimb nerve afferents to external cuneate nucleus of the rat as revealed by intraneural injection of a neurotoxic lectin, Ricinus communis agglutinin. AB - This study seeks to extend the observations of previous studies of projection of primary afferent fibres from the forelimb nerves and muscles to the external cuneate nucleus (ECN) of mammals using a neurotoxic lectin, Ricinus communis agglutinin (RCA) to achieve chemical ganglionectomy of the dorsal root ganglia. Following intraneural injection of RCA into the three main forelimb nerves, namely the radial, ulnar and median nerves, terminal degeneration of the primary afferent fibres in the ECN was studied under the light microscope by means of the Fink-Heimer method. The results show that the primary afferent fibres from these three nerves project to the medial part of the ECN. The field of terminal degeneration take a crescentic form. The projection from the median nerve was most dorsally located whereas that from the radial nerve was the most ventral with extensive overlaps between them. Of the three nerves, the projection from the radial nerve was the most dense. Rostrocaudally, the three nerves also show extensive overlaps. The rostrocaudal extent of maximum terminal degeneration was greatest for the radial nerve and least for the median nerve. Analysis of variance showed that these differences were statistically significant. This suggests that the radial nerve has the most extensive projection to the ECN and the median nerve the least. PMID- 8720456 TI - Cross-reactive and species specific Mycobacterium tuberculosis antigens in the immunoprofile of Schaumann bodies: a major clue to the etiology of sarcoidosis. AB - Sarcoidosis, once thought to be a variant of tuberculosis, is currently listed as a disease of unknown etiology. The present study was initiated by unpublished observations that Schaumann bodies-the laminated inclusions often encountered in sarcoid granulomas-cross-reacted with commercial polyclonal antibodies to Mycobacterium bovis, Mycobacterium duvalii and Mycobacterium paratuberculosis. Given the broad cross-reactivity of many mycobacterial antigens, those findings lacked specificity but warranted in depth probing of the immunoprofile of the bodies, particularly for specific mycobacterial antigens. Formalin-fixed tissue from eight patients with an established diagnosis of sarcoidosis was studied with panels of antibodies against both common cytoplasmic proteins and various mycobacterial antigens, using a labeled streptavidin-biotin-alkaline phosphatase technique. Our findings indicate that Schaumann bodies are indeed residual bodies of heterophagic mycobacterial derivation. They immunostained intensely for the lysosomal proteins muramidase and CD68, variably for some cytoskeletal proteins (tubulin, desmin, vimentin) and not at all for cytokeratin, muscle actin, alpha-1 antichymotrypsin and ferritin. Both cross-reactive and species specific antigenic determinants of M. tuberculosis complex were shown to be present. Affinity absorption with killed intact bacilli H37 Rv resulted in virtually equal loss of binding by all polyclonal antimycobacterial antibodies to cross-reactive ligands in Schaumann bodies. In addition, the bodies were clearly labeled with the monoclonal antibodies TB68 and TB71, known to recognize species specific epitopes of Mycobacterium tuberculosis complex. Although obtained on a small number of cases, our findings uphold Schaumann's original postulate that the laminated calcific inclusions represent remnants of "transformed tubercle bacilli". PMID- 8720457 TI - Immunocytochemistry of perinatal rat livers with a special reference to the roles of mesenchymal cells in hepatic differentiation. AB - To investigate the roles of extracellular matrix produced by hepatic mesenchymal cells in the organization of hepatic cell cords, perinatal rat livers were examined with immunocytochemistry of fibronectin (FN) and laminin (LM). Some hepatocytes in a free state at prenatal day 15 actively produced FN and LM in the rough endoplasmic reticulum but lost this synthetic activity when such cells were incorporated into hepatic cell cords. On the other hand, hepatic mesenchymal cells, especially those associated with the perisinusoidal space, retained this synthetic activity throughout the stages examined. In the differentiating hepatic cell cords, positive immunoreactions for FN and LM were preferentially seen on the cell surface facing both sinusoidal space and differentiating bile canaliculus concomitant with the expression of the tight junction protein, ZO-1, from prenatal day 17. Since such hepatocytes have lost or reduced their synthetic activities of both glycoproteins in the rER, the immunoreactions appear to be mainly due to hepatic mesenchymal cells which seem to play a role in the formation of the hepatic cell cords and the bile canaliculi. PMID- 8720458 TI - Immunohistochemistry of new type I alveolar epithelial cell markers of the rat. AB - The presence of pan-cadherin and the MEP-I antigen in normal and diseased rat lung was established by employing immunoperoxidase and double label fluorescence techniques. The binding of a mouse monoclonal antibody (MEP-1) reacting specifically with type I pneumocytes was assessed on paraffin sections of normal specimens as well as those with pulmonary fibrosis induced by bleomycin or radiation treatment. In injured alveolar epithelium, a diminished type I cell and a focal type II cell immunoreactivity was found. Electron microscopy of immunogold-labelled lung tissue confirmed the type I cell specificity of MEP-1. In severely injured pulmonary parenchyma MEP-1-negative areas occurred, which were also negative with the type II pneumocyte marker Maclura pomifera lectin. Similarly, a polyclonal pan-cadherin antibody uniformly decorated luminal surfaces of alveoli except the type II pneumocytes. Furthermore, pleural mesothelial cells, bronchiolar epithelial cells, endothelial cells of large blood vessels and alveolar macrophage surfaces exhibited pan-cadherin immunoreactivity. After injury, a remarkable loss of pan-cadherin immunoreactivity in the MEP-1 positive type I epithelial cells was detectable. These findings suggest that characterization of normal alveolar epithelial cells and monitoring of the epithelial remodelling in pulmonary pathohistology are sufficiently described by the antibodies MEP-1 anti pancadherin. PMID- 8720459 TI - Immunogold analysis of antioxidant enzymes in common renal cancers. AB - Immunogold studies of normal human kidney and common human kidney cancers were performed using polyclonal antibodies to antioxidant enzymes, including antibodies to copper, zinc and manganese superoxide dismutases, catalase, glutathione peroxidase, and glutathione S-transferases and their subunits. Normal tissue adjacent to human renal tumors had the same antioxidant enzyme immunoreactive protein profiles as normal human kidney, thus establishing that the presence of tumor does not alter the levels of antioxidant enzyme immunoreactive proteins in adjacent kidney tissue. Levels of immunoreactive protein for antioxidant enzymes were determined in four common types of malignant renal cancer. In general, tumors had low levels of antioxidant enzymes; however, certain histologic types of renal tumors had high levels of immunoreactive protein for glutathione S-transferase subunits, which could affect their susceptibility to chemotherapy. Studies of transitional carcinoma of the renal pelvis were especially informative since it was possible to compare levels of antioxidant enzyme immunoreactive protein with adjacent normal transitional epithelium; the majority of antibodies resulted in lower levels of immunoreactive protein in transitional cell carcinoma than in adjacent normal transitional epithelium. Our results are discussed in relation to the response of renal tumors to therapy. PMID- 8720460 TI - Histopathological changes in the islets of Langerhans in hamsters infected with the 139H strain of scrapie: semi-thin section study. AB - Using histopathological analysis of semi-thin sections stained with toluidine blue, we observed profound pathological changes in the islets of Langerhans of hamsters infected with the scrapie agent (strain 139H). These included cytoplasmic vesicles, nuclear swelling, and vacuolization in the islet cells. Two types of vacuolization were seen. "Localized vacuolization" (LV) has a distinct edge and is restricted or confined within the cell. "Diffuse vacuolization" (DV) has no distinct edge and is scattered within tissues either inside or outside of cells. DV may span intracellular and extracellular regions of the islet tissues. There were abnormal structures which we termed blood vessel cores (BVCs) in the islets of 139H-infected hamsters. BVC is a hollow space filled up with blood cells. Immunocytochemical staining for insulin antibody suggested that BVC was surrounded by the B cells of the islet. In the present study, we observed that many inflammatory cells passed through the blood-tissue barriers using pathways between cell-junction in the lumen of BVC. We also observed many necklace-like hollow spaces between islet cells. They are the pockets of extracellular space. A novel concept of "the accordion effect" was described to explain a function of the extracellular space. Under normal physiological conditions, as the synthesis of insulin increase in B cells, the volume of the B cells will increase while the volume of the extracellular space will decrease. After a synchronized secretory response from the stimulated B cells, the secretory product would move from the intracellular space into the extracellular space, the volume of the B cells would be decreased and the volume of the extracellular space would be increased. Most of the secretory product might be released into the blood stream immediately, causing an insulin releasing peak in the blood stream, whereas the rest would remain in the enlarged extracellular space. As the cycle repeat, the increasing volume of the B cells will squeeze the remaining insulin into the blood stream gradually. Thus, the expandable extracellular space would serve as buffer system and a reservoir to collect and store some secretory products for future use. We refer to this concept as "the accordion effect". The concept of "the accordion effect" may also be true in other endocrine organs such as pituitary gland and adrenal gland. PMID- 8720461 TI - Lysosomal glycolipid storage in the renal tubular epithelium in mastomys (Praomys coucha). AB - The renal proximal tubular epithelium of MCC strain of mastomys (Praomys coucha) exhibited a number of cytoplasmic vacuoles after conventional paraffin-embedding procedures. These vacuoles were strongly PAS-positive in cryostat sections. Ultra structurally, they were double membrane-bound structures filled with myelin figures and acid phosphatase-positive electron-dense matrix. Immunofluorescent microscopy revealed that these structures contained GM2 ganglioside. Other tissues or organs were histologically normal. Mating experiments indicated that the ganglioside storage in MCC mastomys is inherited as an autosomal recessive trait. PMID- 8720462 TI - Dynamic interactions of the extracellular matrix. AB - The extracellular matrix (ECM) is a dynamic assemblage of interacting molecules that reorganise and regulate cell functions in response to endogenous and exogenous stimulii. Matrix components may affect cell behaviour directly or indirectly through growth factor sequestration and transmembrane signalling, by controlling the speed of various molecules through the ECM, and the access of growth factors, hormones and neurotransmitters to the cell surface. PMID- 8720463 TI - Ultrastructure of invertebrate muscle cell types. AB - The muscular cells of invertebrates can be divided into three major classes on the basis of their striation pattern: transversely striated, obliquely striated, or smooth muscle. Transversely striated muscles have either continuous or discontinuous Z lines and, thus, can be subdivided into two types respectively. Of all invertebrate muscles, the transversely striated muscle with continuous Z lines is the most similar to the vertebrate skeletal muscle and is present in arthropods, whose musculature (including the visceral muscles) only consists of this cell type. These muscles are multinucleate cells that contain myofibrils showing well-defined sarcomeres. Transversely striated muscles with discontinuous Z lines, consisting of multiple small electrondense patches, are found in the translucent portions of adductor muscles of some bivalves and in the heart muscle of the gastropods. This muscle is formed by mononucleated cells with centrally located nuclei and a single myofibril. The obliquely striated muscle appears in nematodes, annelids, molluscs, brachiopods and chaetognathes and consists of mononucleated cells with both thick and thin myofilaments which form sarcomeres delimited by Z lines. Myofilaments are not perpendicular but oblique to the Z lines, so that both A and I bands may be seen together in each of the three spatial planes of view. Smooth muscle has been reported in coelenterates, annelids, molluscs, brachiopods and echinoderms, but is lacking in arthropods. These muscle cells have a centrally-located nucleus and abundant thin and thick myofilaments without apparent sarcomeres. The most relevant characteristics of invertebrate muscle cells are the following. The thick (myosin) myofilaments show a variable length (from 2.2 microns up to 6 microns) and width (from 14 nm up to 231 nm) and contain a central core of paramyosin, which is absent in vertebrate muscles. Thick filaments are homogenous in transversely striated muscles and either homogeneous or fusiform in the obliquely striated and smooth muscles. Thin filaments measure 6 nm in diameter. They contain tropomyosin and, only in striated muscles, also troponin. The thin/thick filament ratio varies from 3/1 to 6/1, even in smooth muscles. The plaques for filament anchorage (Z lines in striated muscles or electrondense bodies in smooth muscles) contain alpha actinin. The striated (transversely or obliquely) muscles show long sarcomeres (up to 9 microns) and the number of thin filaments around each thick filament varies from 3 to 12, so that each thin filament is shared by two thick filaments. Z lines in the striated muscles show a variety of structures that differ from one species to another (filament bundles in nematodes, bars in annelids, small patches in molluscs, etc). Many striated muscles contain titin (connectin) and intermediate filaments and display a sarcotubular system consisting of T tubules and sarcoplasmic reticulum tubules. Both structures form dyads and, more rarely, triads. The location of T tubules as well as the configuration and distribution of sarcoplasmic reticulum vary among muscles and species. Invertebrate smooth muscle differs from that of vertebrates principally in the higher proportion and larger diameter of thick myofilaments. These may be fusiform and their size and number may vary widely among cells. These muscle cells may be classified by the characteristics of both the thick filaments and the electrondense bodies for filament anchorage. PMID- 8720464 TI - Histochemical demonstration and analysis of poly-N-acetyllactosamine structures in normal and malignant human tissues. AB - Poly-N-acetyllactosaminyl structures carry a variety of physiologically and pathologically important carbohydrate antigens and are presumed to have essential roles in the process of cellular recognition, differentiation, malignant transformation and cancer metastasis. Monoclonal antibodies, lectins and endo beta-galactosidase are useful histochemical tools for detecting and analyzing poly-N-acetyllactosamines in tissue sections. I (branched structure) and i (linear structure) antigens recognized by monoclonal antibodies have been shown to be differentiation antigens in mouse embryo and mouse and human teratocarcinoma cells as well as in human erythrocytes. They are also oncofoetal antigens and are expressed in carcinoma cells in several tissues and organs. Immobilized lectins specific to poly-N-acetyllactosamine structures have been successfully applied for fractioning glycoproteins with poly-N-acetyllactosamine, but histochemical use of these lectins has been restricted to some animal tissues. Among them, pokeweed mitogen agglutinin was used to detect branched poly N-acetyllactosamine in normal and malignant human colon, demonstrating that it has a highly selective affinity for colorectal carcinomas. Griffonia simplicifolia agglutinin-II staining following endo-beta-galactosidase digestion procedure revealed the presence of poly-N-acetyllactosamine structures with or without blood group-specificities in several normal human tissues. By using this procedure, it was demonstrated that the blood group-related antigens oncofoetally expressed in thyroid carcinoma cells are carried by poly-N-acetyllactosamines containing a domain susceptible to the enzyme digestion. Staining with lectins specific to poly-N-acetyllactosamine in combination with endo-beta-galactosidase digestion demonstrated that poly-N-acetyllactosaminyl structures ubiquitously and consistently produced in thyroid papillary carcinomas are highly heterogeneous in their chain length and branching status and quite different from those produced in other thyroid neoplasms. Staining with monoclonal antibodies or lectins combined with endo-beta-galactosidase digestion procedures have been proven to be powerful tools for localizing and analyzing different types of poly-N acetyllactosamine structures in normal and malignant tissues. PMID- 8720465 TI - Cell rounding with "rip off" detachment. AB - The "rounding up" characteristic of mitotic, apoptotic, transformed and non transformed cells appears to be initiated by various causative factors. Cytoskeletal and adhesion modulations have been implicated. Endocytic internalization via large channels seen in the rounding of amoeba and human cells alike, implicate a third mechanism, viz. membrane flow and fusion, where "rip off" detachment sheds parts of the cell to achieve freedom. Mutilation as a means for detachment is also used by moving fibroblasts, linking shape change with movement via a distinct mechanism. While rounding may follow detachment and detachment may follow rounding, "rip off" mutilation, whether self-inflicted or imposed, and modulation of stickiness are fundamentally different processes. PMID- 8720466 TI - Tissutal imaging by nuclear magnetic resonance. AB - The present work reviews the main applications of nuclear magnetic resonance (NMR)-technology and, in particular, of magnetic resonance imaging (MRI) to tissutal analysis. To date, MRI represents a precise and reliable tool to investigate morphology and functional modification of tissues in vivo, providing information consistent with histology. MRI has numerous advantages over conventional techniques: it is harmless to tissues; volume measurements in vivo could be useful for morphometric studies; the same tissue can be examined several times (e.g. at different ages); several organs can be examined at the same time; serial sections of relevant structures can be obtained in all planes, thereby allowing detailed reconstruction of the three-dimensional configuration of organs; motion within a tissue can be detected; and subsequent histological and ultrastructural studies of the tissue are possible. The main drawback (besides the cost of the basic instrumentation) is that resolution is relatively low in comparison with light microscopy. Finally, the analysis of the results is difficult, needing interdisciplinary competence, and MRI methods of tissutal analysis are not yet well standardized. Therefore, in our opinion, MRI is an interesting tool, complementary to other histological techniques, and it cannot be ignored by microscopists. However, in vivo MRI data must be evaluated with caution and histological controls are always required. PMID- 8720467 TI - Elimination of transformed cells by normal cells: a novel concept for the control of carcinogenesis. AB - Control of transformed cells by neighbouring normal cells is known since the beginning of transformation studies in vitro. The classical explanation for this phenomenon is based on proliferation inhibition of transformed cells by normal cells. We extend this model by presenting data that show that TGF-beta-treated normal cells can eliminate transformed cells by induction of apoptosis. Both the TGF-beta-induced signal pathway in normal cells, leading to the production of a short-lived apoptosis-inducing factor, as well as the specific interaction of this factor with transformed cells depend on the action of reactive oxygen species. Sensitivity to induction of apoptosis seems to be a common feature associated with the transformed state, independent of the originally transforming principle. Therefore, tumor development should require either interference with the process of elimination or acquisition of resistance against it. We discuss experimental evidence for interfering substances, such as antioxidants, as well as for genetic systems that protect transformed cells from the negative effects of their cellular environment, such as Bcl-2 or papilloma viruses. These findings, as well as the general resistance of exvivo tumor cells against induction of apoptosis are in line with the novel model of control of tumor progression presented by us in this review. PMID- 8720468 TI - Expression of renin in coagulating glands. AB - The presence of an extrarenal or local reninangiotensin system has been noted in several tissues, although their functions have not yet been clarified. Renin from the coagulating gland (CG), is the most recently discovered local renin, and is a significant subject for investigation with histological and molecular biological techniques. Dot-like reactions for renin are detected immunohistochemically in the epithelial cells of CGs of the strains C57BL/6 mice. Excretory products of some terminal lumina are also found to be positive for renin. Colloidal gold particles, indicating the presence of renin, are detected in the lysosomal granules, in which they are especially located on the crystalline structure. They are also observed in the production of series of exocrine granules. At the apical region, both reninpositive exocrine and lysosomal granules are secreted by exocytosis. In the development, immunoreactivity for renin is first detected at 6 weeks after birth. After that time, the number of renin-containing cells gradually increase throughout the development. In adults, several patterns of renin immunoreactivity are demonstrated in almost all epithelial cells of CGs. At 4 weeks after castration, renin-containing cells in terminal ducts are decreased and remain at very low levels. After testosterone injection, the numerical value of renin-containing cells is high at 1 week. Finally, renin mRNA is detected in the CGs by Northern blot analysis and hybridohistochemistry. These findings suggest that renin is synthesized depending on testosterone, and released by exocrine secretion. PMID- 8720469 TI - Principles of MHC class I-mediated antigen presentation and T cell selection. AB - Class I molecules of the major histocompatibility complex (MHC) are expressed on the cell surface of almost all nucleated mammalian cells. Their main function is to transport and present peptides, derived from intracellularly degraded proteins, to cytotoxic T cells (CTL). They are also directly involved in the process leading to maturation and selection of a functional CD8+ T cell repertoire. MHC class I molecules consist of a highly polymorphic membrane spanning heavy chain of approximately 45 kD that is non-covalently associated with a light chain, beta 2-microglobulin (beta 2m). Class I molecules bind peptides, usually 8-11 amino acids in length. The majority of the class I-bound peptides are generated in the cytosol and are subsequently translocated into the lumen of the endoplasmic reticulum (ER) through the ATP-dependent transporter associated with antigen processing 1/2 (TAP1/2). Here, we provide an up-to-date review summarizing the most essential parts relating to MHC class I-mediated antigen processing, presentation and T cell selection. A particular emphasis is devoted to the structure of MHC class I molecule, and MHC class I-bound peptides. PMID- 8720470 TI - Learning deficits induced by chronic intraventricular infusion of quinolinic acid -protection by MK-801 and memantine. AB - The NMDA receptor agonist quinolinic acid (9 mM) was infused i.c.v. via ALZET osmotic minipumps for 2 weeks. This treatment produced a persistent, short-term memory deficit in the T-maze. Autoradiography revealed a decrease in the density of choline uptake sites in the hippocampus. Parallel s.c. infusion by another minipump of the uncompetitive NMDA receptor antagonist memantine (1-amino-3,5 dimethyladamantane, 20 mg/kg per day) or (+)-5-methyl-10,11-dihydro-5H dibenzocyclohepten-5,10-imine maleate ((+)-MK-801, 0.31 mg/kg day) prevented the learning deterioration induced by quinolinic acid. The treatment with memantine resulted in steady-state serum levels of 1.2 mu M which, based on in vitro data, should assure inhibition of NMDA receptors and are similar to levels seen in the serum of demented patients treated with this agent. In naive animals this treatment had no effect on either learning or on ex vivo induction of long-term potentiation, indicating that under chronic conditions it is possible to obtain neuroprotective effects with NMDA receptor antagonists without negative effects on memory processes. This contrasts to some acute insults (e.g. ischaemia) where high doses of NMDA receptor antagonists that produce side effects are required. PMID- 8720471 TI - Antinociceptive and behavioral effects of synthetic deltorphin analogs. AB - A possible correlation of behavioral, antinociceptive and cataleptic responses with central delta- and mu-opioid receptor stimulation was tested for in the rat by i.c.v. injections of some synthetic deltorphin analogs. At doses ranging from 0.1 to 3.0 nmol/rat, the selective delta-opioid receptor agonist, [D Ala2,Glu4]deltorphin (Tyr-D-Ala-Phe-Glu-Val-Val-Gly-NH2), induced a dose dependent stereotyped pattern of locomotor activity, reaching the maximum in the first 30 min; doses higher than 30 nmol induced early and fleeting antinociception. The replacement of Glu4 by Gly, Ala, Val, His or Asn yielded peptides with a lower delta-selectivity because of a gain in mu-affinity. [D Ala2,Ala4]deltorphin (0.14-4.0 nmol) induced negligible behavioral stimulation but a rapidly appearing and long-lasting analgesia and catalepsy. The other four synthetic peptides induced biphasic effects: low dosages stimulated locomotion whereas higher doses initially suppressed, then increased locomotor activity. At doses ranging from 1 to 70 nmol all the peptides induced analgesia and catalepsy. In experiments examining the locomotor and antinociceptive effects induced by 14 nmol of [D-Ala2,Gly4]deltorphin in rats pretreated with mu and delta antagonists, the non-selective mu-opioid receptor antagonist, naloxone (1 mg/kg i.p.), reduced analgesia and abolished the initial hypolocomotion. The delta-selective antagonist, naltrindole (10 mg/kg i.p.), abolished locomotor activity without affecting analgesia. The mu1 -selective antagonist, naloxonazine (10 mg/kg i.v.), seemed to prolong analgesia and immobility. Hence this peptide appears to activate, in addition to delta-receptors, mainly the opioid receptor mu2-subtype, which mediates catalepsy in the rat. We suggest that the mu2- and delta-opioid receptors of the rat brain modulate locomotor behavior by activating functionally opposed responses. [D-Ala2,Ala4]deltorphin had an antinociceptive and cataleptic potency higher than would have been expected from its mu-affinity. A possible explanation might be a mu/delta-opioid receptor interaction. PMID- 8720472 TI - Modulation of opioid analgesia by agmatine. AB - Administered alone, agmatine at doses of 0.1 or 10 mg/kg is without effect in the mouse tailflick assay. However, agmatine enhances morphine analgesia in a dose dependent manner, shifting morphine's ED50 over 5-fold. A far greater effect is observed when morphine is given intrathecally (9-fold shift) than after intracerebroventricular administration (2-fold). In contrast to the potentiation of morphine analgesia, agmatine (10 mg/kg) has no effect on morphine's inhibition of gastrointestinal transit. delta-Opioid receptor-mediated analgesia also is potentiated by agmatine, but kappa1-receptor-mediated (U50,488H; trans-3,4 dichloro-N-methyl-N-[2-(1-pyrrolidinyl)cyclohexyl] benzeneacetemide) and kappa3 opioid receptor-mediated (naloxone benzoylhydrazone) analgesia is not significantly enhanced by any dose of agmatine tested in this acute model. In chronic studies, agmatine at a low dose (0.1 mg/kg) which does not affect morphine analgesia acutely prevents tolerance following chronic morphine dosing for 10 days. A higher agmatine dose (10 mg/kg) has a similar effect. Agmatine also blocks tolerance to the delta-opioid receptor ligand [D-Pen2,D Pen5]enkephalin given intrathecally, but not to the kappa3-opioid receptor agonist naloxone benzoylhydrazone. Despite its inactivity on kappa1-opioid analgesia in the acute model, agmatine prevents kappa1-opioid receptor-mediated tolerance. These studies demonstrate the dramatic interactions between agmatine and opioid analgesia and tolerance. PMID- 8720473 TI - Central antinociceptive effect of L-ornithine, a metabolite of L-arginine, in rats and mice. AB - L-Arginine produces central antinociception by acting as a precursor of kyotorphin (L-tyrosyl-L-arginine), a [Met5]enkephalin releaser. This study investigated the antinociceptive activity of L-ornithine, a metabolite of L arginine. L-Ornithine given s.c. at 300-1000 mg/kg suppressed carrageenin-induced hyperalgesia in rats in a naloxone-reversible manner. L-Ornithine and L-arginine, when given i.c.v. at 10-100 mu g/mouse, elicited antinociception even in intact mice, the effects being abolished by naloxone or naltrindole, and potentiated by bestatin, an inhibitor of aminopeptidase and kyotorphinase. The antinociception induced by i.c.v. L-ornithine was also inhibited by i.c.v. L-leucyl-L-arginine, a kyotorphin receptor antagonist, but was resistant to intracisternal anti kyotorphin serum. L-Tyrosyl-L-ornithine, a synthetic dipeptide, (1-10 mu g/mouse, i.c.v.), exerted kyotorphin-like antinociception in mice. These findings suggest that L-ornithine produces L-arginine-like antinociception via kyotorphin receptors. However, this effect does not appear to be mediated by kyotorphin itself, but most likely by L-tyrosyl-L-ornithine, a putative dipeptide. PMID- 8720474 TI - Melatonin analogues as agonists and antagonists in the circadian system and other brain areas. AB - We studied the effects of drugs related to melatonin on neuronal firing activity in the suprachiasmatic nucleus, intergeniculate leaflet and other brain areas in urethane-anesthetized Syrian hamsters. We tested melatonin and two naphthalenic derivatives of melatonin, a putative agonist (S20098: N-[2-(7-methoxy-1 naphthyl)ethyl]acetamide), and a putative antagonist (S20928: N-[2-(1 naphthyl)ethyl]cyclobutyl carboxamide). Both melatonin and S20098 given intraperitoneally (i.p.) were able to suppress firing rates of cells in a similar dose-dependent manner, but the effects of S20098 were longer lasting. Iontophoresis of melatonin dose dependently depressed spontaneous and light evoked activity of cells in the suprachiasmatic nucleus and intergeniculate leaflet, while iontophoresis of S20098 was relatively ineffective, probably because it is a poorly charged compound. S20928 (2.0-10 mg/kg, i.p.) alone decreased firing rates of light-sensitive cells by 25-50% for 5-30 min in the suprachiasmatic nucleus and intergeniculate leaflet; however, low doses (< 2.0 mg/kg) of S20928 partially blocked the effects of melatonin agonists on most cells. The non-selective serotonin antagonist metergoline did not block the effects of either melatonin agonist. Both melatonin agonists and antagonists were less effective when applied to cells in the hippocampus and dorsal lateral geniculate nucleus. These results indicate that S20098 is an agonist acting probably on melatonin receptors in the Syrian hamster brain. S20928 may have mixed agonist/antagonist properties, but at low doses appears to function as an antagonist at melatonin receptors in the suprachiasmatic nucleus and intergeniculate leaflet. PMID- 8720475 TI - Chronic ethanol exposure selectively increases diazepam-insensitive [3H]RO15-4513 binding in mouse cerebellum. AB - The effect of chronic ethanol exposure and withdrawal on [3H]RO15-4513 (ethyl-8 azido-5,6-dihydro-5-methyl-6-oxo-4H-imidazo[1,5a][1,4] benzodiazepine-3 carboxylate) binding to diazepam-sensitive and diazepam-insensitive binding sites was determined in mouse brain. Neither chronic ethanol treatment nor withdrawal significantly altered total [3H]RO15-4513 binding in mouse cortex or cerebellum. However, diazepam-insensitive [3H]RO15-4513 binding density (Bmax) in cerebellum was significantly increased immediately following chronic ethanol treatment (60%) and at 8 h following withdrawal (75%). [3H]RO15-4513 binding affinity was not significantly influenced by chronic ethanol exposure or withdrawal. These results indicate that chronic ethanol treatment and withdrawal can selectively up regulate diazepam-insensitive [3H]RO15-4513 binding sites and suggest that this unique GABAA receptor subtype may play some role in ethanol dependence and withdrawal. PMID- 8720476 TI - Methoxamine-induced inhibition of the positive inotropic effect of endothelin via alpha1-adrenoceptors in the rabbit heart. AB - The influence of methoxamine on the positive inotropic effect of endothelin was assessed in the isolated rabbit ventricular myocardium. Methoxamine by itself elicited a positive inotropic effect and it simultaneously inhibited the positive inotropic effects of endothelin-1 and endothelin-3 without affecting the acceleration of the hydrolysis of phosphoinositide that was induced by the endothelin isopeptides. By contrast, the positive inotropic effects induced by elevation of concentration of external Ca2+ ions, by Bay k 8644 (methyl 1,4 dihydro-2,6-dimethyl-3-nitro-4-(2-trifluoromethylphenyl)-pyridine-5- carboxylate), by dihydroouabain and by forskolin were unaffected by methoxamine. The inhibitory action of methoxamine was abolished by alpha1-adrenoceptor antagonists, such as prazosin, WB 4101 (2-(2,6-dimethoxyphenoxyethyl)aminomethyl 1,4-benzodioxane hydrochloride) and (+/-)-tamsulosin; and it was inhibited to a lesser extent by chlorethylclonidine. In addition, methoxamine did not modify the specific binding of [125I]endothelin-3 to ventricular membrane fraction. These results indicate that methoxamine antagonizes the positive inotropic effect of endothelin isopeptides at the level of the signal-transduction process, subsequent to acceleration of the hydrolysis of phosphoinositide, via activation of alpha1-adrenoceptors in the rabbit ventricular myocardium. PMID- 8720477 TI - Cardiovascular responses mediated by two types of endothelin ET(B) receptor in spontaneously hypertensive and Wistar-Kyoto rats. AB - This study shows the effects of a selective endothelin ET(B) receptor agonist, IRL 1720 {Ac-[Ala11,15]endothelin-1-(8-21)}, on cardiovascular responses in anesthetized spontaneously hypertensive rats and Wistar-Kyoto rats. Single intravenous bolus injection of IRL 1720 caused a dose-related short-lasting fall in blood pressure, left ventricular pressure and myocardial contractility. However, repeated intravenous bolus injection of 10(-5) mol/kg IRL 1720 produced a biphasic response consisting of an initial short-lasting decrease followed by a sustained increase in these parameters. The initial decrease was reduced, whereas the following increase was enhanced with the repeated injections of IRL 1720. The cardiovascular pressor response was not inhibited by the endothelin ET(A) receptor antagonist, FR139317 ((R)2-[(R)-2-[(S)-2-[[1-(hexahydro-1H azepinyl)]carbonyl] amino-4-+methylpentanoyl] amino-3-[3-(1-methyl-1H indolyl)]propionyl]amino -3- (2-pyridyl)propionic acid). The effects of IRL 1720 were qualitatively similar but more potent in spontaneously hypertensive rats than in Wistar-Kyoto rats. These results suggest the existence of two types of endothelin ET(B) receptor for IRL 1720: a tachyphylactic endothelin ET(B) receptor that mediates cardiovascular depressor responses and a less tachyphylactic endothelin ET(B) receptor that mediates pressor responses in the rat. PMID- 8720478 TI - Pharmacological characterization of cardiovascular responses induced by endothelin-1 in the perfused rat heart. AB - The effects of the endothelin receptor antagonist TAK-044 (cyclo[D-alpha-aspartyl 3-[(4-phenylpiperazin-1-yl)carbonyl]-L-ala nyl-L-alpha-aspartyl-D-2-(2 thienyl)glycyl-L-leucyl-D-tryptophyl]+ ++disodiu m salt) and BQ-123 (cyclo[D-Asp Pro-D-Val-Leu-D-Trp]) were studied in the rat heart to characterize the receptor subtypes responsible for the cardiovascular actions of endothelin-1. Endothelin-1 induced a transient decrease and subsequent increase in perfusion pressure in perfused rat hearts, and increased left ventricular developed pressure. TAK-044 diminished these endothelin-1-induced responses (100 pmol/heart) with IC50 values of 140, 57 and 1.3 nM, respectively. BQ-123 (1-30 mu M) partially inhibited the endothelin-1-induced hypertension (30-40%) in the rat heart, and failed to inhibit the hypotension. The positive inotropic effect of endothelin-1 was abolished by BQ-123. Neither indomethacin (10 mu M) nor Nomega-nitro-L-arginine methyl ester (100 mu M) attenuated the endothelin-1-induced hypotension. TAK-044 and BQ-123 attenuated the positive inotropic effect of endothelin-1 in rat papillary muscles. In rat cardiac membrane fractions, TAK-044 and BQ-123 inhibited [125I]endothelin-1 binding to endothelin ET(A) receptors with IC50 values of 0.39 +/- 0.6 and 36 +/- 9 nM, respectively, whereas only TAK-044 potently blocked the endothelin ET(B) receptor subtype (IC50 value: 370 +/- 180 nM). These results suggest that endothelin-1 modulates cardiovascular functions in the rat heart by activating both endothelin ET(A) and endothelin ET(B) receptors, all of which are sensitive to TAK-044. PMID- 8720479 TI - Reversal of in vitro lipopolysaccharide-induced suppression of contraction in rat aorta by NG-nitro-arginine, diphenyleneiodonium and di-2-thienyliodonium. AB - The effects of NG-nitro-L-arginine (L-NNA), D-NNA, diphenyleneiodonium and di-2 thienyliodonium on contraction were studied in endothelium-denuded rat aortic rings incubated for 4 h with lipopolysaccharide (10 mu g ml-1) or vehicle. Lipopolysaccharide reduced Emax and increased EC50 of the phenylephrine (10-9-10 5 M) curve. Addition of D-NNA (4, 6 x 10-4 M), L-NNA (1, 10 x 10-6 M) and diphenyleneiodonium (1, 3 x 10-7 M), but not di-2-thienyliodonium (10-7 M), increased Emax and reduced EC50 of the phenylephrine curve of lipopolysaccharide incubated but not control rings. Therefore, D-NNA, L-NNA and diphenyleneiodonium, but not di-2-thienyliodonium, inhibit inducible NO synthase in vascular smooth muscles. PMID- 8720480 TI - The effect of inhaled K+ channel openers on bronchoconstriction and airway microvascular leakage in anaesthetised guinea pigs. AB - Since orally administered K+ channel openers may have cardiovascular side effects, it is possible that inhaled administration would be preferred for the treatment of asthma. We have investigated whether inhaled levcromakalim and HOE 234 inhibit histamine-induced bronchoconstriction and airway plasma exudation in anaesthetised guinea pigs. We have also investigated whether inhaled HOE 234 inhibits the bronchoconstriction and plasma exudation induced by vagus nerve stimulation, which is due to the release of tachykinins from sensory nerves. Lung resistance was measured by airway resistance (RL) computed from airway and transpulmonary pressures and plasma exudation by measurement of Evans blue dye extravasation. Inhaled levcromakalim (25 mu g/ml) had a short duration of action, being effective against histamine-induced bronchoconstriction 2 min after pretreatment, but not at 10 min. Inhaled HOE 234 (25 mu g/ml) was similarly effective against histamine-induced bronchoconstriction but had a longer duration of action. Inhaled levcromakalim partially attenuated histamine-induced plasma extravasation in small airways, but not in the trachea or main bronchi, whereas inhaled HOE 234 had no effect. HOE 234 protected against non-adrenergic non cholinergic nerve-induced bronchoconstriction, but had no effect on neurogenic- or substance P-induced plasma extravasation in the airway. Inhaled K+ channel openers protect against induced bronchoconstriction, but provide little or no protection against plasma exudation, possibly because of an increase in airway blood flow. In addition, inhaled HOE 234 had no effect on neurogenic leakage, suggesting that its vagal inhibitory effect on bronchoconstriction was on airway smooth muscle, rather than on release of neuropeptides from sensory nerves. PMID- 8720481 TI - Epithelial modulation of cholinergic responses in rabbit trachea is partly due to neutral endopeptidase activity. AB - By the simultaneous measurement of acetylcholine release and smooth muscle contraction in rabbit tracheal segments with and without epithelium, pre- as well as postsynaptic effects of this cell layer were studied on cholinergic neurotransmission. The epithelial cell layer exerted a presynaptic inhibitory influence on acetylcholine release, induced by KCl and electrical stimulation, with a concomitant decrease in the smooth muscle contractions. The responses elicited by exogenous acetylcholine, acting postsynaptically, were also inhibited in the presence of the epithelium. The epithelial effect was not accounted for by the production of inhibitory prostaglandins or a nitric oxide-synthase product. Furthermore, the epithelium did not function as a metabolic site for the degradation of acetylcholine. Phosphoramidon, an inhibitor of neutral endopeptidase, mimicked the effects of epithelium removal on the cholinergic responses to high frequency stimulation and on the acetylcholine-induced effects. Neutral endopeptidase inhibition did not further enhance the responses in epithelium-denuded segments. We therefore suggest that the inhibitory function of the epithelium can be partly explained by the activity of neutral endopeptidase, limiting the excitatory effects of tachykinins on cholinergic responses. An alteration in the neutral endopeptidase activity as a result of inflammatory responses and epithelial damage can contribute to the mechanism of airway hyperreactivity in asthma. PMID- 8720482 TI - His381 of the rat CCKB receptor is essential for CCKB versus CCKA receptor antagonist selectivity. AB - A great interest is devoted to antagonists of the cholecystokinin type B (CCKB) receptor such as L-365,260, which reduces panic attacks in humans and to antagonists of the cholecystokinin type A (CCKA) receptor, such as L-364,718 which might be efficient in mental diseases. The A/B specificity of these antagonists was proposed to be mainly dependent on the amino acid sequence of the seventh transmembrane domain (Mantamadiotis and Baldwin (1994) Biochem. Biophys. Res. Commun. 201,1382). In our study, one of these residues, His381 was replaced in the rat CCKB receptor by leucine (the corresponding residue in the CCKA receptor), phenylalanine or arginine using site-directed mutagenesis. Changing histidine for leucine or phenylalanine did not modify significantly the affinity of the CCKB receptor antagonists, L-365,260 and PD-134,308 although both compounds belong to different chemical classes, but strongly improved the affinity of the CCKA receptor antagonists tested. Interestingly, the A selectivity of these CCKA receptor antagonists was recovered by substituting His381 by arginine. Moreover, these results are discussed on the basis of a three dimentional model of the CCKB receptor. The mutated receptors possessed unchanged binding properties for agonists, suggesting that determinants confering specificity for agonists and antagonists are different. PMID- 8720483 TI - Pyrrolidine dithiocarbamate inhibits immunostimulant-induced tetrahydrobiopterin synthesis in rat vascular smooth muscle. AB - GTP cyclohydrolase I is the first and rate-limiting enzyme in the synthesis of tetrahydrobiopterin, a cofactor of nitric oxide (NO) synthase. Immunostimulants increase NO and tetrahydrobiopterin synthesis in vascular smooth muscle cells by coinducing NO synthase and GTP cyclohydrolase I gene expression. Given that nuclear factor kappa(B) mediates the induction of NO synthase gene expression by lipopolysaccharide (LPS), the role of nuclear factor kappa(B) in the induction of GTP cyclohydrolase I in LPS-stimulated rat vascular smooth muscle cells was assessed by examining the effects of pyrrolidine dithiocarbamate, an inhibitor of the activation of nuclear factor kappa(B), on the abundance of GTP cyclohydrolase I mRNA and biopterin synthesis. Pyrrolidine dithiocarbamate inhibited both NO and biopterin synthesis induced by LPS in a dose-dependent manner with similar half maximal inhibitory concentrations, 12 mu M for NO and 17 mu M for biopterin, respectively. At a concentration of 25 mu M, which inhibited NO and biopterin synthesis but caused no cytotoxicity, pyrrolidine dithiocarbamate substantially reduced the LPS-induced increase in the abundance of NO synthase and GTP cyclohydrolase I mRNAs. These results suggest that pyrrolidine dithiocarbamate inhibits LPS-induced NO and biopterin synthesis by inhibiting the expression of NO synthase and GTP cyclohydrolase I genes. Thus, the induction of both genes necessary for cellular NO synthesis in vascular smooth muscle appears to be regulated, at least in part, by a common mechanism: nuclear factor kappa(B) activation. PMID- 8720484 TI - Selective enhancement of antagonist ligand binding at muscarinic M2 receptors by heparin due to receptor uncoupling. AB - The selectivity of heparin in inducing potentiation of binding of antagonist ligands to muscarinic receptors was investigated at the five known subtypes of muscarinic receptors. The effects of heparin on binding of [3H]N methylscopolamine at equilibrium was studied in Chinese hamster ovary (CHO) cells which express each of the individual muscarinic receptor subtypes and in membranes prepared from these cells. Heparin markedly increased equilibrium binding of subsaturating concentrations of the ligand only in membranes of CHO cells which express muscarinic M2 receptors. These effects of heparin were qualitatively similar to those obtained in heart membranes. In contrast, heparin did not influence ligand binding to muscarinic M2 receptors in intact cells. The positive cooperative effects of heparin at muscarinic receptors were abolished following treatment of cells with pertussis toxin. The latter treatment by itself resulted in a significant increase in [3H]N-methylscopolamine binding. Taken together with previous reports of heparin-induced uncoupling of receptors and G proteins, these data suggest that the effects of heparin on ligand binding to muscarinic M2 receptors might be due to disruption of receptor-G-protein interactions which results in enhancement of binding of antagonist ligands to the receptor. PMID- 8720485 TI - Dopamine and 5-hydroxytryptamine selectively potentiate neuronal type ATP receptor channels. AB - Dopamine and 5-hydroxytryptamine have been shown to facilitate a cationic current activated by extracellular ATP in rat pheochromocytoma PC12 cells. Effects of these and other modulators were examined by expressing ATP receptor channels in Xenopus oocytes using cDNAs from rat vas deferens ('P2x1-purinoceptor channels') and PC12 cells ('P2x2-purinoceptor channels'). Dopamine and 5-hydroxytryptamine (10 and 100 mu M) facilitated the ATP-activated current mediated through P2x2 purinoceptor channels, but not the current through P2x1-purinoceptor channels. Adenosine (1 mu M) facilitated the current through both P2x1- and P2x2 purinoceptor channels. Cd2+ (1 mM) as well as Zn2+ (10 mu M) selectively potentiated the current through P2x2-purinoceptor channels. The results suggest that (1) the facilitation by dopamine and other modulators also occurs in recombinant ATP-receptor channels, and (2) the selective facilitation by dopamine, 5-hydroxytryptamine and divalent cations of P2x2-purinoceptor channels is attributed to some structural difference of the channels from P2x1 purinoceptor channels. PMID- 8720486 TI - Immunohistochemical studies of mesolimbic dopaminergic neurons in Fischer 344 and Lewis rats. AB - Previous work has shown that the inbred Lewis and Fischer 344 rat strains differ in several behavioral measures related to mesolimbic dopamine function. Moreover, the ventral tegmental area (VTA) of the Lewis rat has been shown to contain higher levels of tyrosine hydroxylase compared to that of the Fischer rat by blot immunolabeling procedures. To investigate structural correlates of this biochemical difference, an immunohistochemical study of VTA dopaminergic neurons in these two strains was undertaken. Results show that the density and total number of tyrosine hydroxylase-positive neurons in the VT 4 of the Lewis rat is about 50% of that found in the Fischer rat. In contrast, examination of the substantia nigra in the same sections revealed no differences in the density and number of tyrosine hydroxylase-positive cells between these strains. Fischer Lewis strain differences were also evident for cholecystokinin immunoreactivity in the VTA, with much lower levels seen in the Lewis rat, consistent with the known colocalization of this neuropeptide in many VTA dopamine neurons. The finding of 50% fewer tyrosine hydroxylase-positive neurons in the VTA of the Lewis rat, along with our earlier results showing 45% higher levels of tyrosine hydroxylase by blot immunolabeling, would suggest much higher levels of tyrosine hydroxylase per VTA neuron in this strain. However, no obvious strain difference in the cellular intensity of tyrosine hydroxylase immunoreactivity could be detected by immunohistochemistry. Finally, the density of VTA dopamine neurons was assessed in 1-week-old Fischer and Lewis rats. In contrast to the results obtained for adult animals, no difference in the number of tyrosine hydroxylase positive neurons was apparent in these young animals, indicating that the Fischer Lewis strain difference in VTA dopamine neurons appears later in postnatal development. These anatomical findings shed new light on the differences in the mesolimbic dopamine system between Fischer and Lewis rats that may contribute to the behavioral differences exhibited by these animals. PMID- 8720487 TI - Cannabinoid receptor down-regulation without alteration of the inhibitory effect of CP 55,940 on adenylyl cyclase in the cerebellum of CP 55,940-tolerant mice. AB - The objective of this study was to determine whether the development of tolerance to CP 55,940, a potent cannabinoid agonist, was due to changes in the receptor or second messenger system. ICR mice treated with CP 55,940 (2 mg/kg) twice a day for 6 and one-half days developed a high degree of tolerance to the pharmacological effects of CP 55,940. The ability of CP 55,940 to produce motor hypoactivity, hypothermia and immobility was reduced 163-, 97- and 19-fold, respectively. Evaluation of 3H-CP 55,940 binding to rat brain membranes indicated no difference in receptor affinity between the vehicle- and CP 55,940-treated animals. However, these binding studies revealed a 50% decrease in receptor number in the cerebellum of the CP 55,940-tolerant mice. Although cAMP is generally considered to be the second messenger for cannabinoid receptors, little difference was observed in the inhibitory effects of CP 55,940 on adenylyl cyclase activity in cerebellum between vehicle and drug-treated mice. However, there was an increase in receptor mRNA which suggests a compensation for receptor loss. There are several possible explanation for these results. There may be sufficient spare receptors such that CP 55,940-tolerant mice are capable of producing a maximal effect on the second messenger system. On the other hand, one could conclude that cannabinoid receptor down-regulation does not account for the development of tolerance to all of the effects of CP 55,940 in mice. PMID- 8720488 TI - Effects of portal infusion of hypotonic- and hypertonic solutions on neuronal activity in the rat dorsal motor nucleus of the vagus. AB - The present experiment was designed to elucidate the characteristics of the response of neurons in the dorsal motor nucleus of the vagus (DMV) to stimulation of the hepatoportal area by hypotonic as well as hypertonic solutions. Responses of 81 neurons that exhibited an antidromic response to electrical stimulation of the ventral gastric vagus were recorded in the left DMV in urethane-chloralose anesthetized rats. The effects on these 81 neurons of portal infusion of hypertonic saline (3.6% NaCl) and of pure water were examined. The discharge rates of 16 neurons increased in response both to portal infusion of hypertonic saline and to that of water. Portal infusion of 0.9% NaCl produced no changes in firing rates. Their discharge rates of seven neurons increased in response to portal infusion of hypertonic saline but not to that of water. The other 58 neurons did not respond to these stimuli. Jugular infusion of water produced no response. Therefore, the responses to portal infusion of water appear to be derived from activation of the hepatoportal receptors. These results indicate that a certain fraction of DMV neurons respond similarly to portal infusions of hypertonic and hypotonic saline. It is possible that there exist some reflex arcs that mediate a similar response to both an increase and a decrease in portal blood osmolarity (or Na+ concentration), namely, a suppression of absorption. PMID- 8720489 TI - Gap junctions couple astrocytes but not neurons in dissociated cultures of rat suprachiasmatic nucleus. AB - Individual neurons dissociated from rat suprachiasmatic nucleus can express independently phased circadian firing rhythms in culture. The phases of these rhythms are unperturbed by reversible blockade of neuronal firing lasting 2.5 days, indicating that multiple circadian clocks continue to operate in the absence of conventional synaptic transmission. The possibility remains, however, that these circadian rhythms might depend on some other form of intercellular communication. In the present study, a potential role for gap junctional coupling in SCN cultures was evaluated by introduction of the tracer molecule Neurobiotin into both neurons (n = 98) and astrocytes (n = 10), as well as by immunolabeling for specific connexins, the molecular components of gap junctions. Astrocytes were extensively coupled to each other by connexin 43-positive gap junctions, but no evidence was found for coupling of neurons to each other or to astrocytes. These data support the hypothesis that neurons expressing independently phased circadian rhythms in SCN cultures ('clock cells') are autonomous, single cell circadian oscillators, but do not exclude a role for glia in synchronizing neuronal clock cells in vivo. PMID- 8720490 TI - Amygdaloid input to transiently tyrosine hydroxylase immunoreactive neurons in the bed nucleus of the stria terminalis of the rat. AB - Several studies have reported transient expression of tyrosine hydroxylase in a subpopulation of neurons in the bed nucleus of stria terminalis of preadolescent rats. The tyrosine hydroxylase immunoreactive (TH) neurons, which are of small to medium size and often display a typical bipolar configuration, are confined to the intermediate part of the lateral bed nucleus. By the use of a combination of experimental tracer techniques and immunocytochemical methods, we have demonstrated that these neurons receive a significant number of amygdaloid afferents, which establish mostly symmetric synaptic contacts on the cell bodies and sparsely spined dendritic shafts of the TH neurons. TH neurons also receive a small number of tyrosine hydroxylase-positive terminals of unspecified origin. PMID- 8720491 TI - Characterisation of dopamine receptors in insect (Apis mellifera) brain. AB - In vitro binding experiments using the vertebrate D1 dopamine receptor ligand [3H]SCH23390 and the vertebrate D2 dopamine receptor ligand [3H]spiperone were conducted on membrane preparations of honey bee (Apis mellifera) brain. Specific binding of [3H]SCH23390 was saturable and reversible. Analysis of saturation data gave an apparent Kd of 6.3 +/- 1.0 nM and Bmax of 1.9 +/- 0.2 pmol/mg protein for a single class of binding sites. The specificity of high affinity [3H]SCH23390 binding was confirmed in displacement experiments using a range of dopaminergic antagonists and agonists. The rank order of potency for antagonists was: R(+) SCH23390 > cis-(Z)-flupentixol > or = chlorpromazine > fluphenazine > S(+) butaclamol > spiperone. R(+/-)-SKF38393 and dopamine were the most effective agonists tested. [3H]SCH23390 labels a site in bee brain that is similar, but not identical to the vertebrate D1 dopamine receptor subtype. [3H]Spiperone also bound with high affinity to bee brain homogenates. Scatchard analysis of [3H]spiperone saturation data revealed a curvilinear plot suggesting binding site heterogeneity. The high affinity site had a apparent Kd of 0.11 +/- 0.02 nM and Bmax of 9.2 +/- 0.5 fmol/mg protein. The calculated values for the low affinity site were a Kd of 19.9 nM and Bmax of 862 fmol/mg protein. Kinetic analyses also indicated that [3H]spiperone recognises a heterogeneous population of sites in bee brain. Furthermore, agonist competition studies revealed a phenolaminergic as well as a dopaminergic component to [3H]spiperone binding in bee brain. The rank order of potency of dopaminergic antagonists in competing for [3H]spiperone binding was: spiperone > fluphenazine > S(+)-butaclamol > domperidone > R(+) SCH23390 > S(-)-sulpiride. PMID- 8720492 TI - Distribution of hippocampal cholinergic neurostimulating peptide (HCNP) immunoreactivity in the central nervous system of the rat. AB - Hippocampal cholinergic neurostimulating peptide (HCNP), an undecapeptide isolated from the hippocampal tissue of young rats, enhances the cholinergic development in explant cultures of medial septal nuclei. This report concerns the distribution of HCNP immunoreactivity in the central nervous system (CNS) of 11- and 28-day-old Wistar rats; two affinity-purified anti HCNP antibodies were used. Immunoblot analyses of extracts of different regions of the brain revealed a single 23 kDa band that corresponded to the presumed HNCP precursor protein. Immunostaining of the various CNS structures of the 28-day-old rats was more intense than in those of 11-day-old animals. HCNP immunoreactivity was detected in neurons as well as in glia cells, particularly oligodendroglia. The perikarya of neurons in the cerebral cortex, hippocampus, limbic cortex, caudate, putamen, arcuate nucleus of hypothalamus, trigeminal subnuclei, rostroventrolateral reticular nucleus and dorsal horn of the spinal cord were positively stained. In addition, nerve fibers and terminals in the hypothalamic subnuclei, zona incerta, thalamic subnucleus, caudate, putamen, locus coeruleus, trigeminal subnuclei, dorsal motor nucleus of the vagus, dorsal horn of the spinal cord and intermediolateral column also displayed HCNP immunoreactivity. These observations would suggest that HCNP and its related molecules may have multifunctional roles in the CNS. PMID- 8720493 TI - Expression of interleukin-6 receptor (IL-6R) and gp130 mRNA in PC12 cells and sympathetic neurons: modulation by tumor necrosis factor alpha (TNF-alpha). AB - Recent findings indicate that IL-6, besides its various biological effects, also exerts neurotrophic and neuroprotective functions. Using the pheochromocytoma cell line PC12 and cultured primary sympathetic neurons, we investigated whether neurons express the IL-6 receptors, IL-6R and gp130, and how they might be regulated. For these studies we used RT-PCR and in situ hybridization. We provide here evidence for the expression of functional IL-6Rs in peripheral sympathetic neurons and PC12 cells. Furthermore we demonstrate that cytokines modulate the expression of IL-6R and gp130 mRNA. This modulation is much more pronounced in neuronally-differentiated PC12 cells than in undifferentiated cells. Among various cytokines tested, tumor necrosis factor alpha (TNF-alpha) turned out to be a major regulator of the IL-6R and gp130 mRNA expression. The induction was time- and dose-dependent for both genes. Maximal induction was reached within 16 h at a concentration of 0.1 nM TNF-alpha. The stimulatory effect of TNF-alpha on the IL-6R system was completely inhibited by the simultaneous addition of the glucocorticoid dexamethasone. In summary, our results show that sympathetic neurons and neuron-like differentiated PC12 cells express functional IL-6R and gp130, and that the expression of their mRNAs is modulated by cytokines. We suggest that cytokines such as IL-6 can modulate sympathetic neuron function. PMID- 8720494 TI - Kappa-opioid receptor modulation of the release of substance P in the dorsal horn. AB - Substance P (SP), a member of the tachykinin peptide family, has been found in high concentrations in the superficial laminae of the dorsal horn and it is thought to play a major role in the transmission of nociceptive information. Dynorphin(1-8), an opioid peptide with high selectivity for the kappa-opioid receptor subtype, is also found in the dorsal horn of the spinal cord. The aim of this study was to determine the effect of dynorphin(1-8) on the release of SP like-immunoreactivity (SPLI) in the dorsal horn before and during the activation of peripheral nociceptors by a thermal stimulus. A push-pull canula was used to perfuse the dorsal horn of non-anesthetized decerebrate/spinal transected rats and the collected perfusates were assayed for SPLI by using radioimmunoassay. Dynorphin(1-8) applied to the spinal cord at a concentration of 1 microM elicited a 27 +/- 8% decrease in the basal release of SPLI and prevented the increase in the release of SPLI evoked by the application of a noxious thermal stimulus to the ipsilateral hind paw and lower limb. The effect of dynorphin(1-8) was reversed by 2 microM of nor-binaltorphimine (nor-BNI), a selective kappa opioid receptor antagonist. Application of nor-BNI alone to the perfusate resulted in a 62 +/- 23% increase in the basal release of SPLI. In conclusion, dynorphin(1-8) reduces the basal release of SPLI and prevents the increase in the release of SPLI elicited by the application of a noxious cutaneous thermal stimulus. This effect is mediated through the kappa-opioid receptor, which appears to tonically regulate the release of SPLI in the dorsal horn. PMID- 8720496 TI - Pyramidal cells in primary auditory cortex project to cochlear nucleus in rat. AB - Recent work has demonstrated that the auditory cortex in rat sends direct projections to the auditory nuclei of the brainstem, including the cochlear nucleus and superior olive. To determine the cortical origin of the projections to cochlear nucleus, Fast Blue, a retrograde fluorescent tracer, was injected into the cochlear nucleus. Labeled cells in the forebrain were then studied with light microscopy and mapped. The projection was found to originate from large pyramidal neurons in layer V of primary auditory cortex. The projection was predominantly ipsilateral, and no labeled neurons were found in other cortical areas. These data imply that primary auditory cortex exerts influence over ascending auditory information at the earliest stages of the central auditory system. PMID- 8720495 TI - Studies on the identity of the rat optic nerve transmitter. AB - The possible role of glutamate, aspartate, sulfur-containing excitatory amino acids and gamma-glutamyl peptides as major transmitters in the rat optic nerve was evaluated. Four days following optic nerve lesion the K(+)-evoked Ca(2+) dependent glutamate release was reduced to 31 +/- 16% (+/- S.D., n = 9) comparing release from slices of the denervated (contralateral to the lesion) and non denervated (ipsilateral) superior colliculus, indicative of a major transmitter function for glutamate. However, significant decreases in glutamate release could not be detected seven days following the lesion (n = 5). Other studies have shown that optic nerve denervation induce formation of synapses of non-retinal origin and cause other cellular changes which may reduce the effect of deafferentation on glutamate release after 7 days. No significant change was observed in aspartate release following the lesion. The concentrations of cysteine sulfinate, cysteate, homocysteine sulfinate, homocysteate and O-sulfo-serine in the optic layers of the superior colliculus were below 1 nmol/g tissue (n = 6). Theoretical considerations indicate that this level is too low for a function of any of these as a major optic nerve transmitter. All postsynaptic components in the rat superior colliculus response, evoked by electrical optic nerve stimulation, were reduced by kynurenate (1-10 mM), a broad spectrum glutamate-receptor antagonist. The study gives further support for the view that glutamate is a major transmitter in the rat optic nerve. PMID- 8720497 TI - Distribution of Rab3a in rat nervous system: comparison with other synaptic vesicle proteins and neuropeptides. AB - In the present study we have investigated the distribution of Rab3a in rat peripheral nervous system and compared it with the distribution of other synaptic vesicle proteins (synaptophysin, synapsin I), neuropeptides (CGRP, SP, NPY) and tyrosine hydroxylase (TH). Rab3a immunoreactivity (-IR) was always colocalized with synaptophysin-IR and synapsin I-IR in nerve terminals of the spinal cord and peripheral nerve endings. In many cases, Rab3a-IR was also present in the same axons and terminals as peptides. In crushed sciatic nerve axons, Rab3a was colocalized, proximal to the crush, with synaptophysin-IR, synapsin I-IR, CGRP IR, and TH-IR, but only partially co-localized with NPY-IR and SP-IR. In the area distal to the crush, Rab3a-IR was very weakly positive in a few thin axons, while larger amount of synaptophysin, CGRP, NPY and SP immunoreactivities were detected. The subcellular distribution of peptides and Rab3a differed in that peptides were observed mainly in large granular structures, while Rab3a-IR was observed mainly as diffuse, finely granular immunoreactivity, in addition to a few exceptional large granules present in some axons. The results demonstrate that Rab3a is widely distributed in different types of neurons, i.e. motor, sensory, autonomic adrenergic and cholinergic neurons, and colocalized with other synaptic vesicle proteins, suggesting that Rab3a may play an essential role in neuronal function. Furthermore, Rab3a is present in many peptide containing axons and terminals, but with an apparently different subcellular distribution, being affiliated mostly with small synaptic vesicles and only occasionally with large vesicles, that may represent peptide contained vesicles. PMID- 8720498 TI - Coexistence of parvalbumin and GABA in nonpyramidal neurons of the rat entorhinal cortex. AB - The possible coexistence of the calcium-binding protein, parvalbumin, with the major inhibitory neurotransmitter, gamma-aminobutyric acid (GABA), and its synthesizing enzyme, glutamate decarboxylase (GAD), was studied in nonpyramidal cells of the rat medial and lateral entorhinal cortex. The material was analyzed by two different methods, the first of which was a mirror techniques where the possible coexistence of two different antigens was analyzed from cells cut in half at the surface of the adjacent section. The other method consisted of analyzing double immunofluorescent-stained sections with a confocal microscope. The colocalization analysis revealed that all parvalbumin-immunoreactive neurons (mirror technique n = 688 and confocal microscopy n = 644) in all layers of the medial and lateral entorhinal cortex were also immunopositive for GABA or GAD. Parvalbumin-cells made up 52% of the GABA cells in most of the layers in the medial and lateral entorhinal cortex. In layer III of the entorhinal cortex, the proportion was about 40%. Thus, parvalbumin-containing neurons in the entorhinal cortex represent a large GABAergic cell population, which is likely to play an important role in controlling both the input and the output of the entorhinal cortex. PMID- 8720499 TI - Contribution of excitatory amino acids to morphine-induced metabolic alterations. AB - Previous studies have indicated that excitatory amino acids are involved in the analgesic and addictive properties of morphine. However, their role in the morphine-induced alterations in glucose metabolism is not known. This study assessed the contribution of NMDA receptor activation to the morphine-induced hormonal and metabolic alterations in conscious unrestrained chronically catheterized rats. Whole body glucose flux was assessed with a primed constant intravenous infusion of [3-3H]glucose in rats pretreated with the NMDA-receptor antagonist MK-801 (0.25 mg/kg, intraarterial) or an equal volume (1.5 ml) of sterile saline (0.9%) administered 15 min prior to i.c.v. injection of H2O (Con; 5 microliters) or morphine sulfate (80 micrograms). No significant alterations were noted in metabolic and hormonal parameters of H2O injected rats. i.c.v. morphine increased the plasma glucose concentration (60%), hepatic glucose production (Ra; 60%) and whole body glucose utilization (Rd; 53%), but did not alter the glucose metabolic clearance rate (MCR). MK-801 alone resulted in transient hyperglycemia (25%), stimulation of glucose Ra (60%) and glucose Rd (53%), and a significant (30%) increase in MCR. MK-801 pretreatment blunted the morphine-induced hyperglycemia and the increased glucose Ra and Rd. Morphine increased the plasma concentration of epinephrine (4-fold), norepinephrine (2 fold) and corticosterone (67%); however, no alterations in plasma insulin and glucagon were detected. MK-801 pretreatment, blunted the morphine-induced increase in corticosterone and norepinephrine, and elicited a significant rise in insulin concentrations. These results indicate that activation of the NMDA receptors contributes to the morphine-induced hyperglycemia and hormonal alterations. Furthermore, this response appears partially mediated by activation of sympathetic outflow and suppression of insulin release, which is blunted by inhibition of NMDA receptors. PMID- 8720500 TI - Origins of nitric oxide synthase-containing nerve fibers in the rat basilar artery with reference to the fine structure of the nerve fibers. AB - The origins of nitric oxide synthase (NOS)-containing nerve fibers in the rat basilar artery were studied by a combination of Fluoro-Gold retrograde tracing and immunohistochemistry. After application of Fluoro-Gold onto the middle part of the basilar artery, the dye accumulated in the sphenopalatine, otic, trigeminal, superior cervical, nodose ganglia and in the spinal ganglia at level C2 and C3. Nerve cells with NOS-like immunoreactivity were detected in the above ganglia, except for the superior cervical ganglion. Neurons that showed both NOS like immunoreactivity and Fluoro-Gold fluorescence were numerous in the sphenopalatine and otic ganglia, and less numerous in the trigeminal, nodose and spinal ganglia. Under electron microscopy, a number of unmyelinated nerve terminals with neuronal NOS-like immunoreactivity was seen in proximity to smooth muscle cells in the tunica media of the basilar artery. These findings provide morphological evidence that NOS-containing nerve fibers in the rat basilar artery have multiple origins, and suggest that the control of posterior cerebral circulation by the parasympathetic and sensory ganglia are more complex than previously considered. PMID- 8720501 TI - The effect of adrenalectomy on stress-induced c-fos mRNA expression in the rat brain. AB - Previously, we determined the pattern of stress-induced c-fos mRNA expression throughout the brain in order to gain further insight into the identification of the neural circuits mediating stress-induced regulation of the hypothalamic pituitary-adrenal axis. In the present study, we determined if rapid effects of increased glucocorticoid levels after stress contribute to changes in c-fos mRNA expression. To this end, stress-induced c-fos expression was characterized in adrenalectomized (ADX) or adrenalectomized and corticosterone replaced (ADX/B) male rats. Animals were sacrificed 30 min post-onset of a 10 min swim stress, and in situ hybridization histochemistry was used to detect c-fos mRNA throughout the brain. The pattern of c-fos induction in the ADX and ADX/B animals was similar to that observed in the sham operated animals. Additionally, densitometric measurements were made to quantify the c-fos response in the paraventricular nucleus of the hypothalamus and the CA1/2 region of the hippocampus. We found that ADX did not alter the magnitude of the c-fos response to stress in these areas, but there was a slight dampening of the response in ADX/B animals. In sum, these results suggest that the pattern of c-fos expression observed 30 min post stress is independent of stress-induced increases in circulating glucocorticoid concentrations. PMID- 8720502 TI - Augmented pharmacologic stimulation of striatal acetylcholine release following developmental hypoxic-ischemic injury. AB - We have previously shown that developmental hypoxic-ischemic injury in a unilateral rodent model leads to an increase in both morphologic and biochemical indices of striatal cholinergic neurons. To investigate the functional significance these changes, we have used the in vivo microdialysis technique to examine the regulation of striatal acetylcholine release in awake, adult rats following postnatal hypoxic-ischemic injury. We have found that injury does not alter basal release or acetylcholine, but it results in a marked augmentation in the increase of acetylcholine release normally observed after infusion of atropine or pirenzepine. PMID- 8720503 TI - Increase in vasopressin binding sites in the human choroid plexus in Alzheimer's disease. AB - Vasopressin binding sites were determined in the choroid plexus of five Alzheimer's disease patients and five non-demented controls using the 125I labelled linear V1a-antagonist. The Alzheimer's disease patients showed a twofold increase in the density of vasopressin binding sites, whereas the increase in the affinity constant Kd did not reach significance. PMID- 8720504 TI - The effects of a single neonatally induced convulsion on spatial navigation, locomotor activity and convulsion susceptibility in the adult rat. AB - The effect of a single neonatal convulsion on subsequent behaviour was investigated in the male rat. Convulsions were induced by heat or pentylenetetrazol on days 1, 10 or 21. As adults, locomotor activity, spatial ability and convulsion susceptibility were measured. Significant differences were seen in some measures and some groups but a single neonatal convulsion did not induce a consistent and significant pattern of behavioural change, despite the persistent change in hippocampal physiology shown in a previous study. PMID- 8720505 TI - Chronic neurosteroid treatment attenuates single cell GABAA response and its potentiation by modulators in cortical neurons. AB - In previous studies we have observed that chronic neurosteroid 5 alpha-pregnan-3 alpha-ol-20-one (5 alpha 3 alpha) treatment produced downregulation of the GABAA receptors, heterologous uncoupling, and decreased heterologous efficacy at the GABAA receptor complex in cultured mammalian cortical neurons. In this study, using whole cell recording, we examined the consequence of chronic 5 alpha 3 alpha (1 microM; 5 days) treatment on GABA-induced currents in isolated cortical neurons. We observed that the GABA current was decreased by 78% after 5 days treatment of cortical cells with 1 microM 5 alpha 3 alpha. We also observed decreased pentobarbital, and 5 alpha 3 alpha potentiation of GABA currents after chronic 5 alpha 3 alpha treatment. These findings support the notion that GABA response, and its potentiation by pentobarbital, and neurosteroid, 5 alpha 3 alpha, are attenuated after chronic 5 alpha 3 alpha treatment. PMID- 8720506 TI - Correlation between different types of retinal ganglion cells and their projection pattern in the albino rat. AB - Injecting Fluoro-Gold (FG) and Evans-Blue (EB) into the right dLGN and SC in the adult albino rat, ipsilaterally projecting double-labeled retinal ganglion cells were mainly seen in the ventrotemporal crescent. They were mainly large sized cells. The ipsilaterally projecting double-labeled cells tended to have larger somata than the single- and double-labeled cells projecting to the contralateral superior colliculus and/or dorsal nucleus of the lateral geniculate body. PMID- 8720507 TI - Activation of c-Fos contributes to amyloid beta-peptide-induced neurotoxicity. AB - Amyloid beta peptide, a major component of Alzheimer's disease plaques, is directly toxic to various neuronal cell lines and primary neurons in culture. The mechanism underlying A beta neurotoxicity may include an increase in intracellular calcium and reactive oxygen species. In the present study, exposure of a mouse hippocampal cell line (HT-22) to the 25-35 peptide fragment of A beta (10 microM) caused a rapid and sustained increase in nuclear c-Fos immunoreactivity. Inhibition of A beta-mediated c-Fos activation by c-fos antisense oligodeoxynucleotides (5 microM) significantly protected against A beta toxicity as assessed by MTT assay. The signal transduction pathway for c-fos induction remains speculative, however, there seems to be a causal relationship between c-Fos transcription factor and A beta toxicity. PMID- 8720508 TI - Potent and long-lasting anticonvulsant effects of 1-naphthylacetyl spermine, an analogue of Joro spider toxin, against amygdaloid kindled seizures in rats. AB - The anticonvulsant effect of 1-naphthylacetyl spermine (1-NA-Spm), an analogue of Joro spider toxin, against amygdaloid kindled seizures was studied in rats. 1-NA Spm (10, 20 and 40 micrograms/rat) dose-dependently improved kindled seizures and shortened the afterdischarge duration 30 min after the administration. The anticonvulsant effect was observed even one day after the drug, and then gradually disappeared within 4 days. The present findings demonstrate that 1-NA Spm acts as a potent and long-acting anticonvulsant against amygdaloid kindled seizures, and also suggest, together with the previous findings, that the calcium permeable AMPA receptors, which are selectively antagonized by 1-NA-Spm, play a critical role in the seizure generation mechanism of amygdaloid kindling. PMID- 8720509 TI - Aromatic L-amino acid decarboxylase is present in serotonergic fibers of the striatum of the rat. A double-labeling immunofluorescence study. AB - The aim of the present study is to examine whether serotonergic fibers of the striatum of the rat contain aromatic L-amino acid decarboxylase (AADC). By use of a double-labeling immunofluorescence method, we showed that AADC was localized in serotonergic fibers of the striatum and cerebral cortex as well as in serotonergic cell bodies of the midbrain raphe nuclei. We previously demonstrated that serotonergic fibers of the rat striatum contained dopamine after intraperitoneal injection of L-dopa. These findings suggest that dopamine is produced from the injected L-dopa in serotonergic fibers of the rat striatum. PMID- 8720510 TI - Consensual definitions. PMID- 8720511 TI - Media influences on suicidal behavior in young people. PMID- 8720512 TI - Letters across the Atlantic. PMID- 8720513 TI - Elder abuse--a risk factor for suicide? PMID- 8720514 TI - Befriending the suicidal in Yugoslavia. PMID- 8720515 TI - Controversial issues in jail suicide prevention. PMID- 8720516 TI - IASP task force on euthanasia and assisted suicide. AB - The first meeting of the IASP Task Force on Euthanasia and Assisted Suicide took place in Venice on June 7, 1995. Several interested observers were present. It was decided that at the public IASP meeting the following day each speaker should address, briefly, the current legal situation and the pressure for change, as well as give a personal statement. David Clark spoke for North America, Bob Goldney for Australia, Michael Kelleher for Britain and Ireland, Jerzy Wasserman for Scandinavia, and Hans Wedler for the German-speaking world. Their views are published in this article. Ad Kerkhof requested that the Dutch television film "Death on Request" be discussed. The committee was of the opinion that clear definitions were essential. In their view, these should take into account the differences between active and passive euthanasia, as well as between professionally assisted and lay-assisted suicide. PMID- 8720517 TI - Hotline for mental health in Shanghai, China. AB - In this article the author introduces the Hotline for Mental Health in Shanghai. According to the experience gained over the past 2 years, most of the 8214 callers were having difficulties in interpersonal relationships, often in relation to lovers and partners. Love affairs were the main reason for telephone consultation among the unmarried callers (51%), while marital disputes and family problems were the most common reason for married callers (57%). Some people who suffered from mental disorders made calls asking for help in management or social support. These calls included rehabilitation of psychoses (3.5%), sleep disturbance (3.2%), and suicidal ideation (2.2%). The author suggests that such a hotline service can be a simple and practicable crisis intervention approach for suicide prevention in Shanghai. PMID- 8720518 TI - History of early loss among a group of suicide survivors. AB - Suicide-bereaved families have been described as more depression-prone and their bereavement process as more complicated and difficult. Few studies have aimed specifically to investigate the history of transgenerational childhood loss in suicide survivor families as an indicator of greater vulnerability. Childhood separation experiences are associated with difficulties in forming and maintaining attachments. Bonding difficulties are also associated with greater vulnerability to depression and may constitute a suicide risk factor. This exploratory study (n = 32) focused on two groups, each of 16 mothers who had lost a son to suicide and car accidents, respectively. History of loss and early separation experiences were investigated through a clinical interview of the bereaved mothers and through a psychological autopsy of the deceased sons. Results indicate significant transgenerational indices of loss, separation, and inadequate child-rearing. Pre-death life events and adversities in the family may be as important to bereavement outcome as post-death happenings. The cycle of these events, the role of fathers and the role of protective factors must be further studied using detailed life-course reconstructions. PMID- 8720519 TI - Mental health profiles, suicidal behavior, and community sexual assault in 2112 Canadian adolescents. AB - 1025 females and 1087 males in grades 7-12 in Alberta high schools completed measures of emotional and behavioral problems and suicidal behaviors, and of frequency of sexual assaults outside of school. Both males and females experiencing a high number of sexual assaults in the previous 6 months were significantly more likely to have clinical profiles on measures of conduct disorder, somatic disorder, and emotional disorder. 13.2% of 53 girls with frequent, unwanted sexual contact had made more than two suicidal gestures or suicide attempts in the previous 6 months, compared with 1.3% of 783 girls with no experience of sexual assault. 33.3% of 18 boys experiencing frequent sexual assault were suicidal, compared with 0.9% of 1064 boys with no recent experience of sexual assault. These results are in line with results of other recent Canadian surveys. PMID- 8720520 TI - A study on suicide. PMID- 8720521 TI - Conference report: 18th IASP Congress in Venice. PMID- 8720522 TI - Stengel Research Award 1995. PMID- 8720523 TI - Erwin Ringel Award 1995. PMID- 8720524 TI - Demonstration of insulin resistance in coronary artery disease documented with angiography. AB - OBJECTIVE: To evaluate the relation between insulin resistance and coronary atherosclerosis, insulin sensitivity in lean nondiabetic, normotensive subjects with and without obstructive coronary artery disease (CAD). The correlation between insulin resistance and degree of coronary stenosis was also investigated. RESEARCH DESIGN AND METHODS: Four groups were studied: 1) nine subjects with normal glucose tolerance (NGT) without CAD, 2) 10 subjects with NGT with CAD, 3) nine subjects with impaired glucose tolerance (IGT) without CAD, and 4) 10 subjects with IGT with CAD. Insulin sensitivity was determined by the steady state plasma glucose (SSPG) method using Sandostatin. Coronary angiography was performed in all study subjects, and the severity of coronary artery atherosclerosis was quantified in a modified Gensini score. RESULTS: The SSPG (millimoles per liter) levels were significantly higher in the patients with CAD compared with control subjects (control vs. patient group: 4.8 +/- 0.5 vs. 7.9 +/ 0.9 with NGT, P < 0.05; 5.6 +/- 0.5 vs. 11.1 +/- 0.8 with IGT, P < 0.001), indicating the presence of insulin resistance in patients with CAD. The coronary atherosclerosis score (CAS) was significantly and positively correlated with SSPG (r = 0.74, P < 0.05) and 2-h insulin area (r = 0.78, P < 0.01) in NGT subjects with CAD. On the other hand, the percentage fall of plasma free fatty acid (0-30 min) during an insulin sensitivity test was significantly decreased in the subjects with CAD and was inversely correlated with the CAS (r = -0.43, P < 0.05), especially in NGT subjects with CAD. CONCLUSIONS: These data suggest that in patients with CAD, insulin-mediated glucose metabolism is significantly impaired, and a significant correlation was noted between insulin resistance and severity of CAD. Therefore, the hyperinsulinemia often observed in patients with CAD is attributable to the compensatory mechanism of the beta-cell to the inadequate action of insulin for glucose metabolism. Hyperinsulinemia in the presence of insulin resistance aggravates dyslipidemia and may stimulate the atheromatous process by an as-yet-unknown mechanism. PMID- 8720525 TI - Age-related alteration of pancreatic beta-cell function. Increased proinsulin and proinsulin-to-insulin molar ratio in elderly, but not in obese, subjects without glucose intolerance. AB - OBJECTIVE: To determine the secretion of insulin, C-peptide, and proinsulin after oral glucose loading in healthy elderly subjects compared with middle-aged subjects with and without obesity and with NIDDM. RESEARCH DESIGN AND METHODS: Subjects fell into four groups: nonobese middle-aged normal control subjects (CNT group; n = 38, 40-64 years old); obese normal subjects (OB group; n = 18, 40-64 years old); nonobese NIDDM subjects (NIDDM group; n = 28, 40-64 years old); and nonobese elderly subjects (OL group; n = 17, 65-92 years old). Insulin, C peptide, and proinsulin were determined by radioimmunoassay in plasma samples taken at 0, 30, 60, and 120 min during a 75-g oral glucose tolerance test (OGTT). RESULTS: There were no differences in plasma glucose during the OGTT among the three nondiabetic groups. Hyperinsulinemia was significant in the OB and NIDDM groups but not in the OL group. On the other hand, absolute hyperproinsulinemia was significant in the OL and NIDDM groups compared with the CNT group. Increased proinsulin was rather dominant in the OL group, especially late after glucose loading. Molar ratios of proinsulin to insulin or C-peptide thus were significantly higher in the OL and NIDDM groups. CONCLUSIONS: Alteration of pancreatic beta-cell function independent of that seen with NIDDM occurred in relation to aging. This may be a predisposing factor to the development of impaired glucose tolerance or NIDDM in elderly subjects, that is, independent of obesity. PMID- 8720526 TI - Variability in diagnostic evaluation and criteria for gestational diabetes. AB - OBJECTIVE: To determine the frequency of screening for gestational diabetes mellitus (GDM) among a population receiving regular prenatal care and to assess the extent to which National Diabetes Data Group (NDDG) criteria for the diagnosis of GDM are used by practicing obstetricians. RESEARCH DESIGN AND METHODS: We studied participants in the Nurses' Health Study II, a large prospective cohort study of 116,678 nurses aged 25-42 years in 1989. A total of 422 women who reported a first diagnosis of GDM between 1989 and 1991 were sent supplementary questionnaires regarding diagnosis and treatment, and medical records were requested for a subset of 120 to validate self-reported GDM and assess criteria used for diagnosis. A sample of 100 women who reported a pregnancy not complicated by GDM were sent questionnaires addressing GDM screening and prenatal care. RESULTS: Among a sample of 93 women who reported a pregnancy not complicated by GDM and responded to the supplementary questionnaire, 16 (17%) reported no glucose loading test; 69% of unscreened women had one or more risk factors for GDM. Among a sample of 114 women who self reported GDM in a singleton pregnancy and whose medical records were available for review, a physician diagnosis of GDM was confirmed in 107 (94%). Records and supplementary questionnaires indicated that oral glucose tolerance tests (OGTTs) were performed in 96 (86%) of these women. Of women with a physician diagnosis of GDM whose OGTT results were available, 25% failed to meet NDDG criteria for this diagnosis, although all had evidence of abnormal glucose homeostasis. CONCLUSIONS: Screening for GDM is not universal, even among a group of health professionals in whom screening prevalence is likely to be higher than in the general population. Diagnostic criteria for GDM among obstetricians in practice remain nonstandard despite NDDG recommendations. Better understanding of the implications of differing degrees of glucose intolerance and of varying GDM screening and management strategies is required to make policy recommendations for appropriate and cost-effective care. PMID- 8720527 TI - Increased molar proinsulin-to-insulin ratio in women with previous gestational diabetes does not predict later impairment of glucose tolerance. AB - OBJECTIVE: To evaluate if an increased proinsulin-to-insulin ratio (PI/I) in former gestational diabetes mellitus (GDM) subjects could be a marker for later impairment of glucose tolerance. RESEARCH DESIGN AND METHODS: This study is a prospective follow-up. In a previous follow-up study of former GDM subjects 3-4 years after an index pregnancy, an increased PI/I was found also in normoglycemic nonobese former GDM subjects compared with control subjects. A 75-g oral glucose tolerance test (OGTT) was performed 3 years after the first follow-up, i.e., 6-7 years after the index pregnancy in 97 of the former GDM subjects and in 23 control subjects. A 75-g OGTT according to the World Health Organization was performed. Glucose, insulin, proinsulin, and C-peptide were determined at 0, 30, 60, 90, 120, 150, and 180 min after the glucose intake. RESULTS: Since the first follow-up, an additional 3 in 97 (3.1%) and 15 in 97 (15.5%) of the former GDM subjects had NIDDM or impaired glucose tolerance (IGT), respectively. All control subjects still had a normal OGTT. The fasting PI/I at follow-ups 1 and 2 was significantly correlated in the former GDM subjects (r = 0.41, P < 0.001) and in the control group (r = 0.46, P < 0.05). There was no significant correlation between the PI/I in follow-up 1 and the fasting or 2-h glucose values at follow up 2. If GDM subjects with a PI/I in the upper quartile in the first follow-up were compared with those with a lower PI/I, there were no significant differences in outcome of OGTT in the second follow-up. CONCLUSIONS: The hypothesis that an increased fasting PI/I is a marker for later development of NIDDM or IGT in former GDM subjects could not be supported. PMID- 8720528 TI - Abnormalities of antroduodenal motility in type I diabetes. AB - OBJECTIVE: In the present study, a recently developed manometric technique was used to study antroduodenal motility in ambulant type I diabetic subjects. RESEARCH DESIGN AND METHODS: In 12 patients with type I diabetes, antroduodenal manometry was performed for 20 h during the fasting period and the postprandial period after a standardized dinner and breakfast. All patients had evidence of cardiac autonomic neuropathy and complained of dyspeptic symptoms. During the manometric study, the blood glucose levels were frequently monitored and kept close to euglycemia in the diabetic patients. The results were compared with 12 healthy control subjects. RESULTS: The migrating motor complex cycles observed in the diabetic subjects were longer than in the control subjects, 118.9 +/- 46.0 vs. 87.0 +/- 21.6 min (P < 0.05). This increase was attributable to a prolonged phase II, 78.0 +/- 35.5 vs. 37.7 +/- 18.5 min (P < 0.05). In the diabetic subjects, antral phase III was seen significantly less than in the control subjects, 16.7 vs. 43.3% (P < 0.005). In 50% of the diabetic patients, total absence of antral phase III was observed-this phenomenon was not seen in the healthy control subjects. After dinner, the antral motility index was less in diabetic subjects compared with the healthy volunteers, indicating antral hypomotility (P < 0.01). Six diabetic patients showed abnormal duodenal activity such as early recurrence of phase III and bursts after dinner. No significant differences in antral motility index or in duodenal motility patterns were observed after breakfast. Six diabetic patients complained of dyspeptic symptoms after dinner, whereas none had dyspeptic symptoms after breakfast. In 67% of the patients, nausea was reported after an early phase III or a burst. CONCLUSIONS: This study shows that prolonged ambulatory antroduodenal manometry is a feasible technique in patients. Recording multiple migrating motor complexes showed that interdigestive motor abnormalities of the stomach and duodenum are common in diabetic patients. Furthermore, it shows the occurrence of antral hypomotility and abnormal duodenal motility patterns after a high-calorie meal, with dyspeptic symptoms in diabetic patients being related to the composition of the meal. PMID- 8720529 TI - Severe islet amyloidosis in congenital generalized lipodystrophy. AB - OBJECTIVE: Islet amyloidosis may be one mechanism for pancreatic islet beta-cell loss that is associated with the development of NIDDM. However, the question remains whether chronic overstimulation of insulin and islet amyloid polypeptide (IAPP) secretion in states of insulin resistance could lead to formation of islet amyloidosis and hence NIDDM in some patients. We studied pancreatic islet pathology in congenital generalized lipodystrophy, a genetic syndrome of extreme insulin resistance that may provide some clues. RESEARCH DESIGN AND METHODS: Our patient was a 24-year-old African-American woman with congenital generalized lipodystrophy. Severe acanthosis nigricans was noted in her since age 6. At ages 12 and 16, normal and impaired glucose tolerances, respectively, were noted on oral glucose tolerance tests but were accompanied by extreme fasting and post prandial hyperinsulnemia. Overt diabetes developed at age 18 and she required approximately 180 U of insulin daily. Immediately after an accidental death at age 24, an autopsy was performed. Pancreatic histology was studied in detail using routine methods and immunohistochemical techniques. RESULTS: Some scarring of the pancreas as a result of previous episodes of acute pancreatitis was observed. Severe amyloidosis was noted in 89% of the islets, sparing those that were rich in pancreatic polypeptide-secreting cells. Amyloid deposits stained intensely on immunostaining with antibodies against amylin. Marked paucity of beta-cells was evident. The ratio of beta- to alpha-cells was reduced to 1:1 (normal ratio approximately 4:1). CONCLUSIONS: These observations suggest that chronic presence of extreme insulin resistance may induce premature and severe islet amyloidosis as well as beta-cell atrophy. PMID- 8720530 TI - Erythrocyte sodium-lithium countertransport activity in non-nephropathic diabetic twins. AB - OBJECTIVE: It has proved difficult to separate the role of the diabetic state as distinct from its complications in causing the elevation in erythrocyte sodium lithium (Na-Li) countertransport activity that has been observed in diabetes. The present study sought to isolate the impact of diabetes on the countertransporter by studying groups of non-nephropathic identical-twin pairs both discordant and concordant for diabetes. RESEARCH DESIGN AND METHODS: We studied erythrocyte Na Li countertransport activity in 49 identical-twin pairs who were discordant for IDDM and 26 identical twin pairs who were concordant for IDDM. Similar numbers of healthy control subjects, matched with the nondiabetic twins from the discordant pairs in respect to sex, BMI, and age were also studied. RESULTS: The clinical and laboratory characteristics of both sets of twins were very similar to those of the control subjects with the exception that whole-blood glucose and glycated hemoglobin concentrations were higher in diabetic twins, whether from discordant or concordant pairs (P < 0.001), and that systolic blood pressure (P < 0.05) and serum HDL cholesterol (P < 0.05) were higher in the discordant diabetic twins than in their nondiabetic co-twins. Median (95% CI) Na-Li countertransport activities (in millimoles of lithium released from 1 liter of erythrocytes per hour) in the nondiabetic discordant twin [0.237 (0.192-0.284)], the diabetic discordant twin [0.284 (0.254-0.326)], and the concordant twin [0.262 (0.207 0.358)] groups were similar to each other and higher than in the control subjects [0.172 (0.138-0.203)]. Countertransport activities in the discordant diabetic twins correlated significantly with their nondiabetic co-twins (r = 0.34; P = 0.015; n = 49), as did those between the concordant diabetic twin pairs (r = 0.68; P < 0.005; n = 26); activity levels were not related to either disease duration or blood glucose control. CONCLUSIONS: An elevation in Na-Li countertransport activity has been noted in non-nephropathic normotensive twin pairs both discordant and concordant for IDDM. The potential genetic contribution to the altered behavior of the countertransporter was similar in both types of twins studied, and individual Na-Li countertransport activities were not significantly related to either duration of diabetes or metabolic control. PMID- 8720531 TI - Hyperinsulinemia in nondiabetic Asian subjects using specific assays for insulin, intact proinsulin, and des-31, 32-proinsulin. AB - OBJECTIVE: To investigate the contributions of intact proinsulin and of des-31,32 proinsulin to fasting concentrations of insulin-like molecules in nondiabetic subjects from two ethnic groups (Asian and white) and to see whether Asian subjects are hyperinsulinemic compared with white subjects using highly specific assays for insulin. RESEARCH DESIGN AND METHODS: We investigated subjects with normal glucose tolerance (NGT) (82 Asian and 67 white) and impaired glucose tolerance (IGT) (16 Asian and 13 white), diagnosed by using standard World Health Organization criteria. Highly specific monoclonal antibody-based assays were used to measure insulin, intact proinsulin, and des-31,32-proinsulin. An index of insulin secretion was derived as a ratio of incremental insulin to incremental glucose concentrations from 0 to 30 min during an oral glucose tolerance test. RESULTS: Asian subjects with NGT, despite being significantly thinner than whites (BMI 24.4 +/- 3.5 vs. 25.7 +/- 3.7 kg/m2, P = 0.04), had a more central distribution of obesity (subscapsular-to-triceps skinfold ratios 1.36 +/- 0.69 vs. 1.17 +/- 0.41, P = 0.047). Asian subjects with NGT showed significant hyperinsulinemia 2 h after oral glucose load (plasma insulin median 274 pmol/l [range 26-1,505] vs. 186 pmol/l [27-720], P < 0.005) compared with whites. Asian subjects with NGT also had significantly higher insulin increments (P < 0.02) compared with white subjects and significantly higher fasting concentrations of intact proinsulin (median 2.7 pmol/l [range 0.9-14.1] vs. 2.1 [0.8-7.9], P < 0.02) but not of des-31,32-proinsulin. The ratio of proinsulin-like molecules to the total sum of three insulin-like molecules, however, was similar between Asian and white subjects with NGT and IGT. CONCLUSIONS: These results indicate that when specific assays for insulin are used, Asian subjects show postglucose load hyperinsulinemia and fasting hyperproinsulinemia compared with white subjects, suggesting increased insulin secretion and/or the presence of underlying insulin resistance in this ethnic group. The contribution of proinsulin-like molecules to total insulin-like molecules was similar between Asian and white subjects with NGT and IGT, and there was no contribution to hyperinsulinemia in Asian subjects. PMID- 8720532 TI - Prevalence and risk factors for micro- and macroalbuminuria in an Italian population-based cohort of NIDDM subjects. AB - OBJECTIVE: To determine the prevalence of micro- and macroalbuminuria in NIDDM and their relationship with some known and putative risk factors. RESEARCH DESIGN AND METHODS: Out of a population-based cohort of 1,967 NIDDM subjects, 1,574 were investigated (80%). Albumin excretion rate (AER) was evaluated on an overnight urine collection, and plasma and urine determinations were centralized. RESULTS: The prevalences of microalbuminuria (AER 20-200 micrograms/min), macroalbuminuria (AER > 200 micrograms/min), and hypertension were 32.1% (95% CI 29.8-34.4), 17.6% (15.7-19.5), and 67% (64.6-69.3), respectively. Apart from prevalence of hypertension, which after adjustment for age, BMI, and duration of diabetes was 2.3 times higher in women, rates were higher in men (odds ratio [OR] 1.31, 95% CI 1.04-1.66 for microalbuminuria and OR 1.63, 1.22-2.17 for macroalbuminuria). In comparison with normoalbuminuric subjects, both micro- and macroalbuminuric diabetic subjects had significantly longer duration of diabetes, higher levels of systolic blood pressure, fasting plasma glucose, HbA1c, triglycerides, and uric acid; in macroalbuminuric subjects only, levels of apolipoprotein B and HDL cholesterol were, respectively, higher and lower than in normo- and microalbuminuric subjects. In logistic regression, variables independently related to both micro- and macroalbuminuria were age, HbA1c, cigarette smoking habits, plasma uric acid, and diastolic blood pressure, after adjustment for plasma creatinine and diabetic treatment. In addition, duration of diabetes and HDL cholesterol levels were associated with macroalbuminuria. CONCLUSIONS: This population-based study showed high prevalence of micro- and macroalbuminuria in NIDDM subjects, who were characterized by a more adverse pattern of cardiovascular risk factors. PMID- 8720533 TI - Variation in the incidence and proportion of diabetes-related amputations in minorities. AB - OBJECTIVE: To identify the age-adjusted and level-specific incidence of amputations associated with diabetes in Hispanics, African-Americans, and non Hispanic whites. RESEARCH DESIGN AND METHODS: We used a database from the Office of Statewide Planning and Development in California that identified all hospitalizations for lower-extremity amputations in the state in 1991. Amputation level was defined by ICD-9-CM codes 84.11-84.18 and were categorized as toe, foot, leg, and thigh amputations. RESULTS: The age-adjusted incidence of diabetes related amputation per 10,000 persons with diabetes in 1991 was 95.25 in African Americans, 55.98 in non-Hispanic whites, and 44.43 in Hispanics. Hispanics had a higher proportion of amputations (82.7%) associated with diabetes than did African-Americans (61.6%) or non-Hispanic whites (56.8%) (P < 0.001). African Americans had the highest age-adjusted incidence rate for each level in people with and without diabetes. African-Americans underwent more proximal amputations compared with non-Hispanic whites and Hispanics (P < 0.001). Diabetes-related amputations were 1.72 and 2.17 times more likely in African-Americans compared with non-Hispanic whites and Hispanics, respectively. CONCLUSIONS: Hispanics had proportionally more amputations associated with diabetes than did African Americans or non-Hispanic whites. A significant excess incidence of both diabetes and non-diabetes-related amputations and proportionally more proximal amputations were identified in African-Americans compared with Hispanics and non Hispanic whites. A possible explanation could be the higher prevalence of peripheral vascular disease in African-Americans. Public health initiatives, which have been demonstrated to reduce the incidence of diabetes-related lower extremity amputations, should be implemented, and additional work should focus on minority groups. PMID- 8720534 TI - Polymorphism of complement C4 and susceptibility to IDDM and microvascular complications. AB - OBJECTIVE: The aim of this study was to investigate whether or not the inherited polymorphism of complement C4 is associated with genetic susceptibility to microvascular complications in IDDM as previously reported. RESEARCH DESIGN AND METHODS: We determined C4 phenotypes in 241 patients with IDDM and 140 healthy control subjects by agarose gel electrophoresis and immunoprecipitation. C4 allotype frequencies were compared between patients and healthy control subjects. In addition, we compared allotype frequencies of 83 patients with nephropathy with those of 80 patients without nephropathy and compared those of 50 patients with proliferative retinopathy with those of 68 patients without retinopathy or background retinopathy. Duration of IDDM in control patients was at least 21 years. RESULTS: Patients and healthy control subjects differed at both the C4A (P < 0.00001) and C4B (P < 0.0005) loci. The C4 null allele C4AQ0 was significantly increased in IDDM patients (26.8 vs. 11.8%, P < 0.005). C4B2 was more frequently observed in patients (14.5 vs. 6.8%, P < 0.05) compared with healthy control subjects. No differences were observed in C4 allotype distribution between patients with and without nephropathy or retinopathy. CONCLUSIONS: These data confirm previous reports of an association between the C4 null allele C4AQ0 and IDDM. Our results do not support an association of the inherited polymorphism of complement C4 with genetic susceptibility to microvascular complications in patients with IDDM. PMID- 8720535 TI - Can caretakers of children with IDDM accurately measure small insulin doses and dose changes? AB - OBJECTIVE: To determine if caretakers of young children with IDDM could consistently reproduce small incremental measurements of insulin (U100). RESEARCH DESIGN AND METHODS: Fifteen caretakers of children with IDDM were asked to deliver repeated small doses of insulin, including doses separated by only 0.25 U of insulin. A sensitive gravimetric technique was used to determine the error in measurement of these low doses of insulin. Statistical analysis was used to evaluate accuracy and internal consistency of each caretaker at each dose. RESULTS: The means +/- SD at each dose level were as follows: 2.75 +/- 0.13 U at 2.5 U, 3.19 +/- 0.13 U at 3.0 U, 3.55 +/- 0.13 U at 3.25 U, and 3.70 +/- 0.11 U at 3.5 U. All doses were biased toward overadministration. There was as statistically significant difference in the dose delivered when the target doses were varied by only 0.25 U. The average differences and standard errors between 2.5 U and 3.0 U, 3.0 U and 3.25 U, and 3.25 U and 3.5 U were 0.44 +/- 0.20 U, 0.36 +/- 0.018 U, and 0.15 +/- 0.017 U, respectively. CONCLUSIONS: Participants were not accurate in measuring small insulin doses, consistently overdrawing insulin by an average of 0.22 U. Caretakers are reasonably internally consistent with a given dose, since participants were able to measure statistically significant differences in 0.25 U dose changes. The error in insulin measurement does not vary with the intended dose level. Caretakers in the same family deliver insulin doses as variable from each other as they are from the population as a whole; however, when two or more individuals are responsible for one insulin dose in a child with IDDM, they have a combined variability that is approximately 40% greater than a single individual's variability. PMID- 8720536 TI - Islet cell antibodies at diagnosis, but not leanness, relate to a better cardiovascular risk factor profile 5 years after diagnosis of NIDDM. AB - OBJECTIVE: To evaluate the relationship between islet cell antibodies (ICAs) and the cardiovascular risk profile 5 years after clinical diagnosis of NIDDM. RESEARCH DESIGN AND METHODS: Five years after clinical diagnosis, we evaluated blood pressure (BP) and lipids in 17 NIDDM patients with ICA at diagnosis (age 60 +/- 4 years) and 133 NIDDM patients without ICA at diagnosis (age 61 +/- 1 year). Urinary albumin excretion was evaluated in a subset of 12 NIDDM patients with ICA at diagnosis (age 60 +/- 4 years) and 82 NIDDM patients without ICA at diagnosis (age 61 +/- 1 year). RESULTS: NIDDM patients without ICA showed higher BP (140/86 +/- 2/1 mmHg vs. 128/79 +/- 3/2 mmHg; P < 0.05), total cholesterol (6.10 +/- 0.11 vs. 5.09 +/- 0.29 mmol/l; P < 0.01), LDL-to-HDL ratio (3.85 +/- 0.14 vs. 2.49 +/- 0.18; P < 0.001), and triglycerides (2.58 +/- 0.24 vs. 0.90 +/- 0.06 mmol/l; P < 0.001), lower HDL cholesterol (1.08 +/- 0.03 vs. 1.40 +/- 0.08 mmol/l; P < 0.001), and higher urinary albumin excretion (0.16 +/- 0.06 vs. 0.01 +/- 0.01 g/24 h; P < 0.05) than NIDDM patients with ICA. Among NIDDM patients without ICA, no differences concerning BP or lipids were found between obese and nonobese patients. CONCLUSIONS: ICA at diagnosis of NIDDM is a marker of more favorable cardiovascular risk profile 5 years after clinical diagnosis. PMID- 8720537 TI - The effects of high- and medium-dose metformin therapy on cardiovascular risk factors in patients with type II diabetes. AB - OBJECTIVE: To study the dose response to metformin in type II diabetic patients. RESEARCH DESIGN AND METHODS: Type II diabetic patients with a BMI > 25 were treated with 3,000 mg/day (n = 27), 1,500 mg/day (n = 25), or placebo (n = 23) for 6 months. Venous blood samples were taken at each visit for plasma glucose and insulin, HbA1c, triglyceride and cholesterol, plasminogen activator inhibitor 1 (PAI-1) antigen and activity, tissue plasminogen activator (tPA), and euglobulin clot lysis time (ECLT). Blood pressure was recorded at each visit. RESULTS: There were no changes in BMI or blood pressure. Blood glucose fell (mean) by 3.6 mmol/l in the high-dose and 0.5 mmol/l in the low-dose group over the 6-month study (P < 0.001 and NS compared with placebo). HbA1c and plasma insulin fell in both treatment groups (HbA1c, P < 0.001; insulin, P < 0.003 and 0.03). There was a fall in triglyceride (P < 0.05) and cholesterol (P < 0.008) with high-dose metformin. PAI-1 antigen and activity fell by approximately 20% of baseline in both treatment groups (PAI-1 antigen high dose, P < 0.01; PAI-1 antigen low dose, P < 0.002: PAI-1 activity high and low dose, P < 0.003). There were significant falls in total tPA in both groups (P < 0.004), but the overall effect was a fall in ECLT (P < 0.03). CONCLUSIONS: The results indicate that metformin has favorable effects on cardiovascular risk factors associated with type II diabetes. The effects on glycemic control and lipids are dose-dependent, while the enhanced fibrinolytic response is independent of the doses used. PMID- 8720538 TI - Nondiabetic Mexican-Americans do not have reduced insulin responses relative to nondiabetic non-Hispanic whites. AB - OBJECTIVE: To study why Mexican-Americans have a threefold increase in NIDDM relative to non-Hispanic whites. The etiology of NIDDM is still controversial, with both insulin resistance and decreased insulin secretion proposed as precursors of NIDDM. RESEARCH DESIGN AND METHODS: We examined possible ethnic differences in fasting insulin (as a marker of insulin resistance) and change in insulin-to-change in glucose ratio (delta I30:delta G30) during the first 30 min after oral glucose ingestion (as a marker of abnormal whites from the San Antonio Heart Study, a population-based study of diabetes and cardiovascular disease. Fasting insulin and delta I30:delta G30 were evaluated as continuous variables. RESULTS: Mexican-Americans had increased insulin concentrations at fasting and 30, 60, and 120 min after an oral glucose load as well as an increased 0- to 30 min increment in insulin and delta I30:delta G30 relative to non-Hispanic whites. These results remained unchanged after adjustment for age, sex, obesity, body fat distribution, and glucose tolerance. CONCLUSIONS: These results suggest that increased insulin resistance rather than decreased insulin secretion is characteristic of nondiabetic Mexican-Americans, a high-risk population for NIDDM. PMID- 8720539 TI - A 5-year (1989-1993) prospective study of the incidence of IDDM in Rome and the Lazio region in the age-group 0-14 years. AB - OBJECTIVE: To provide data on the incidence of IDDM in Rome and the Lazio region evaluated prospectively from 1989 to 1993 for a total of > 5 million subjects younger than 15 years. RESEARCH DESIGN AND METHODS: All patients with newly discovered IDDM diagnosed between 1 January 1989 and 31 December 1993 among residents in Rome and its region were recorded. Primary ascertainment was based in diabetes clinics and specialized hospitals in the region, whereas the secondary independent source was taken from the archives of the region where patients are registered to obtain exemption from paying for medications. RESULTS: We identified 330 new patients with a degree of ascertainment of 85%. Overall the incidence rate of the disease was 7.9 per 100,000 per year (95% CI 7.1-8.8). The incidence was higher in the 5- to 9-year-old age-group (10.4 per 100,000) and in winter (36.2%). The cumulative risk for the disease is on the order of 1.18 per 1,000 subjects < 15 years of age. No significant differences in incidence were observed between boys and girls. There were 14 instances of coma at diagnosis (4.3%), but hyperglycemia without ketonuria was diagnosed in 35% of patients, suggesting an early diagnosis. DISCUSSION: Compared with the other continental Italian regions for which data are available for a single year, the IDDM incidence rate in Rome is similar. We conclude that the IDDM incidence rate in Rome and its region is comparable to that in other Southern European countries and remained stable over the 5-year observation period. PMID- 8720540 TI - Relationship between GAD antibody and residual beta-cell function in children after overt onset of IDDM. PMID- 8720541 TI - Urinary myo-inositol-to-chiro-inositol ratios and insulin resistance. PMID- 8720542 TI - Microalbuminuria in patients with NIDDM: an overview. AB - We have come a long way in our understanding of the epidemiology, pathophysiology, and clinical significance of albuminuria in patients with NIDDM. However, substantial gaps remain to be defined. NIDDM nephropathy is a serious and increasingly burdensome disease for both the diabetic individual and the society at large. Onset of microalbuminuria, an early but common manifestation of NIDDM nephropathy, marks an ominous turn for the NIDDM patient, in whom its development forecasts a grave cardiovascular outcome. Interception of albuminuria with antihypertensive agents such as ACE inhibitors in otherwise healthy NIDDM subjects holds a significant promise but must first await further investigation. PMID- 8720543 TI - Severe autonomic neuropathy treated with subcutaneous insulin infusion. PMID- 8720544 TI - Inpatient management of adults with diabetes. PMID- 8720545 TI - Diabetes in the Egyptian deserts: a very low prevalence. PMID- 8720546 TI - Antiphospholipid syndrome in a patient with type I diabetes presenting as retinal artery occlusion. PMID- 8720547 TI - Acarbose: in search for its real indications in current medical practice. PMID- 8720548 TI - American Diabetes Association scientific sessions, 1995: IDDM: treatment and prevention. PMID- 8720549 TI - Moral crisis. PMID- 8720550 TI - Treatment of acute migraine with sumatriptan--response in 40 consecutive patients. AB - Forty consecutive headache patients self-administered sumatriptan for migraine, diagnosed by the criteria of the International Headache Society (IHS). Eighty percent reported excellent response. Thirty-three percent had recurrence of headache within four to twelve hours, while 67% had no recurrence. Fifty-five percent of the patients reported mild side effects, but only 8% stopped therapy because of adverse reactions. No serious cardiovascular events occurred. Recommendations for safe use of sumatriptan are suggested. PMID- 8720551 TI - Laser assisted disc decompression: an alternative treatment modality in the Medicare population. AB - The current study evaluated the efficacy of LADD within the Medicare population, and examined the influence of coexisting medical conditions and previous surgical intervention on surgical outcome. Patients who met inclusion criteria underwent laser-assisted disc decompression (LADD) using the Holmium: YAG Laser with Sidefire laser fiber. All patients evidenced primarily leg pain, with or without back pain, which had failed a minimum of six weeks conservative treatment. Post operative follow-up at one year yielded a surgical success rate of 80%. Surgical outcome was not related to coexisting medical conditions or previous surgical intervention on the lumbar spine. Findings of the current study suggest that LADD offers a viable alternative treatment modality for lumbar disc disease within the Medicare population. PMID- 8720552 TI - Surgical treatment of cancer of the pancreas in large community hospitals. AB - A retrospective study compares the success rates of surgical treatment of cancer of the pancreas in large community hospitals. Although none of the surgeons averaged as many as two pancreaticoduodenal resections per year for the period of this study, their results compared well with other published series except for a few centers and surgeons who did a very high volume of such resections. Results of the study indicate that well trained surgeons in well staffed and well equipped community hospitals can provide acceptable results in pancreaticoduodenectomies for cancer of the pancreas. PMID- 8720553 TI - Cardiac transplantation in patients aged 60 years or older. AB - One hundred eighty-four consecutive patients who underwent heart transplantation from January 1987 to December 1994 have been reviewed. Patients who were 60 years of age or older at the time of transplant (Group A, n = 50) showed improved overall survival when compared with younger patients (Group B, n = 134), though this was not statistically significant. Survivals in groups A and B were 94% and 90%, respectively, at 1 year; 86% and 80% at 5 years; and 86% and 78% at 8 years. Overall survival of older patients in the U.S. and worldwide is inferior when compared with younger patients. More thorough pretransplant evaluation of the older patient and improved compliance post-transplantation may be factors in the good results obtained in this group at our own center. PMID- 8720555 TI - Certified athletic trainers discuss best type of ankle support. PMID- 8720554 TI - Risk of "little league elbow" can be reduced, say trainers. PMID- 8720556 TI - Tick-borne illness increasing in Oklahoma. PMID- 8720557 TI - In vitro maintenance of Eperythrozoon suis. AB - In vitro maintenance of Eperythrozoon suis was attempted using a Petri dish erythrocyte culture system. In preliminary experiments, the optimal conditions for maintaining E. suis attachment to erythrocytes during incubation were anticoagulation with heparin or citrate solution, incubation with 5 or 10% CO2 at 37 degrees C, and incubation with reduced or non-reduced Eagle's minimum essential medium. Using heparin, a CO2 incubator and reduced Eagle's medium (rEM), E. suis metabolic activity was evaluated by measuring glucose consumption, and lactate and pyruvate production. Glucose consumption and lactate production were measurable while pyruvate production was not detected. Erythrocyte integrity was improved by the addition of inosine although no effect was observed on maintenance of E. suis attachment to erythrocytes or the rate of glucose consumption. To determine whether the glucose consumption observed in culture was due to E. suis glycolytic activity or enhanced erythrocyte glycolytic activity, the effect of E. suis killing by EDTA addition to medium was evaluated using rEM containing inosine (rEMI). Glucose consumption decreased proportionally with the decline in the percentage of parasitized erythrocytes induced by EDTA, indicating that glucose consumption was due to E. suis. In a subsequent experiment, the effect of different types of serum (pig or fetal calf serum) and different gaseous environments (5% CO2 incubator or candle jar) were evaluated using rEMI. Glucose consumption by E. suis was significantly increased by the addition of fetal calf serum; however, no difference in the maintenance of E. suis attachment to erythrocytes and in E. suis glycolytic activity was observed between a 5% CO2 incubator and a candle jar. Finally, the effect of medium refreshment (rEMI containing fetal calf serum) was evaluated. Maintenance of E. suis parasitism on erythrocytes and E. suis glycolytic activity were significantly improved by frequent medium refreshment. The maintenance system developed enabled successful metabolic radiolabeling of E. suis for protein/antigen analysis. PMID- 8720558 TI - Effect of silica treatment on resistance to Babesia rodhaini infection in immunized mice. AB - The effect of silica treatment on the course of Babesia rodhaini infection was investigated in ICR mice pretreated with either a mixture of B. rodhaini parasitized red blood cell hemolysation-sonication (S) antigen and Propionibacterium acnes bacterin or P. acnes bacterin alone and not subjected to immunization. In the normal mice (non-immunized), the mean survival time was decreased by approximately 1 day when 5 mg per mouse of silica were intraperitoneally inoculated at 5 h before infection. In immunization with S antigen and P. acnes bacterin, all mice survived regardless of the dose of silica (15-25 mg per mouse) treatment. In contrast, in immunization with P. acnes bacterin alone, 3/10, 4/10 and 1/10 mice died of B. rodhaini, when inoculated with 15 mg per mouse, 20 mg per mouse and 25 mg per mouse of silica treatment 5 h before infection, respectively. These results suggest that peritoneal macrophages (M luminal diameter of), especially when activated non-specifically by P. acnes bacterin, play an important role in inhibiting multiplication of organisms at the early stage of infection and the mice immunized with S antigen were ready to quickly acquire an enhanced solid specific protection even if the function of M luminal diameter of was damaged by a high dose of silica treatment. PMID- 8720559 TI - Herd evaluation by enzyme-linked immunosorbent assay for the determination of Fasciola hepatica infection in sheep and cattle from the Altiplano of Bolivia. AB - A study was designed to determine by ELISA the seroprevalence of fasciolosis both in sheep (29 herds totaling 184 sheep), in samples collected in 1988, and in cattle (41 herds totaling 299 animals, samples collected in 1988; 34 herds totaling 147 animals, samples collected in 1989) in the same area of Corapata in which a seroprevalence survey had been done in humans. The results show high seropositivity in sheep (89%) and lower seropositivity in cattle (58% in 1988, and 57% in 1989). The seroprevalence in cattle in 1988 was essentially identical to that detected in 1989. Faecal examinations were also done in the 1988 sheep and 1989 cattle. Results of the study showed that of the 184 sheep examined, 22 were positive for F. hepatica eggs, while 163 were positive by serology. All of the 22 sheep which were positive parasitologically were also positive serologically for a sensitivity of 100%. On the other hand, of 147 cattle tested, 38 were positive parasitologically while 84 were positive serologically. Of the 38 positives for F. hepatica eggs, 31 were positive by serology (sensitivity 82%). PMID- 8720560 TI - Diagnosis of Taenia saginata cysticercosis in Kenyan cattle by antibody and antigen ELISA. AB - Sera from calves, either experimentally or naturally infected with Taenia saginata, were screened for an antibody response to T. saginata, and for parasite antigen, by enzyme-linked immunosorbent assays (ELISAs). An antibody response was detected by 3 weeks post infection (p.i.), rose to a peak at 10-12 weeks p.i., and was still in evidence 1 year p.i. Parasite antigen was first detected 4-7 weeks p.i. and persisted until the end of the experiment, over 1 year p.i. In the experimentally infected animals, cattle with 14 or more live cysticerci had detectable levels of parasite antigen in their sera at slaughter, while animals with live cyst burdens ranging from 0 to 4 were negative. Furthermore, levels of circulating antigen were positively correlated with live cysticercus burden in the experimental animals. In naturally infected cattle, 83% (5/6) of those with 30 or more live cysts, and 22% (5/23) of those with 1-29 live cysts, could be detected by the ELISA for parasite antigen, although no significant correlation between antigen level and live cyst burden could be detected. Antibody levels were not found to be associated with cyst burdens in either experimentally or naturally infected cattle. In slaughterhouse cattle, the antigen assay was almost three times as sensitive as meat inspection. However, there was no agreement between cattle found positive at meat inspection and those found positive by the antigen detection ELISA. One possible reason is that the ELISA only detects live cysts, while lesions left by dead cysts are more noticeable at meat inspection. The mouse monoclonal antibody-based antigen detection ELISA is of value for the diagnosis of naturally occurring, viable, T. saginata cysticercosis in live cattle and has an immediate application for field based epidemiological studies designed to determine prevalence. PMID- 8720561 TI - Comparison of single and trickle infections of pigs with eggs of the Asian Taenia. AB - Two groups of seven Belgian Landrace piglets each were either infected with a single dose of 3000 or with five consecutive doses of 600 Asian Taenia eggs at weekly intervals. Nine weeks after the first infection all pigs were autopsied and the number of metacestodes was obtained by slicing the liver. There were no significant differences between the mean number of viable or dead cysts present in both groups of animals. Only very low numbers of living metacestodes were found: 0.4% (3/779) of the total number of cysts present in the single infection group and 1.8% (13/707) in the trickle infection group. Circulating antigens could be detected in only four out of 13 animals and no differences in antibody kinetics were present between the two groups of pigs. The presence of high numbers of degenerated cysts in experimental as well as in field conditions seems to indicate that the biotic potential of the Asian Taenia is rather low. PMID- 8720562 TI - Use of excretory/secretory antigens for the serodiagnosis of Anoplocephala perfoliata cestodosis. AB - Whole worm extract (WWE) and excretory/secretory (E/S) antigens of Anoplocephala perfoliata were characterised by SDS-PAGE and their use in the serodiagnosis of equine cestodosis was evaluated. An enzyme-linked immunosorbent assay (ELISA) was used to compare WWE and E/S antigen as the capture layer in an antibody capture ELISA. E/S antigen gave the best differentiation between sera from tapeworm positive and tapeworm-negative horses. The E/S-ELISA was optimised and validated against sera from horses of known tapeworm status. This assay gave a diagnostic sensitivity of 68% (n = 38) and a specificity in helminth-naive horses of 95% (n = 20). Western blot analysis of tapeworm-positive and tapeworm-negative sera revealed the immunoreactive proteins of E/S antigen to be those of 12 and 13 kDa. PMID- 8720563 TI - Nematode burdens and immunological responses following natural challenge in Romney lambs selectively bred for low or high faecal worm egg count. AB - Breeding lines of Romney sheep, selected as lambs for consistently low or high faecal nematode egg count (FEC) following periods of natural challenge, have been maintained at Wallaceville for some years. In order to determine the extent to which FECs in low and high genotypes reflected their ability to resist the establishment of gastro-intestinal nematode burdens, we investigated the infection status and immune responses in 8- to 9-month-old progeny of selected rams from low and high FEC breeding lines following a period of grazing without anthelmintic treatment in autumn/early winter. In each of the 2 years of the study, outcross male progeny of the two lowest FEC (LFEC) (i.e. most 'resistant') and two highest FEC (HFEC) (i.e. most 'susceptible') rams from the divergent lines were slaughtered shortly after autumn/early winter FECs had been analysed. Post-mortem worm counts and examination of intestinal histology were then undertaken. Blood samples collected before slaughter in the second year of the study were assayed to measure serum levels of Trichostrongylus colubriformis specific antibody and immunoglobulins (IgG1 and IgM), and numbers of circulating eosinophils. Overall, correlations between pre-slaughter FEC and total trichostrongyle burdens in the lambs proved to be very high (0.91 and 0.85, respectively, for the 2 years studied). In the first year, LFEC lambs, which were shedding only 28.6% as many strongyle eggs as their HFEC counterparts at slaughter, were found to harbour 37.6% as many adult trichostrongyle worms, while in the second year, LFEC lambs, which were shedding 16.1% as many strongyle eggs as their HFEC counterparts at slaughter, were found to harbour 33.5% as many adult trichostrongyle worms. Results, particularly in the second year, confirmed that significantly fewer worms of most of the important abomasal and small intestinal nematode species which infest lambs in New Zealand (i.e. Haemonchus contortus, Ostertagia circumcincta, Cooperia curticei, Nematodirus spathiger, T. colubriformis, and Trichostrongylus vitrinus) had established in the LFEC genotypes than in their HFEC counterparts. In addition, in utero egg counts of female intestinal Trichostrongylus spp. were significantly lower in LFEC lambs than in their HFEC counterparts, indicating a reduction in fecundity of those worms which did establish. There was also some evidence of an effect of host response on the developmental composition of burdens in the case of some worm species. In relation to host responses, numbers of globule leucocytes/mucosal mast cells in the intestinal mucosa were significantly higher (P < 0.01) in LFEC lambs than in HFEC lambs in both years of the study. Numbers of connective tissue type mast cells and eosinophils in the intestinal mucosa were also significantly higher in LFEC lambs but only in the second year of the study (P < 0.01 and P < 0.05, respectively). Numbers of circulating eosinophils did not differ significantly between the genotypes. T. colubriformis-specific antibodies, IgG1 and IgM to both L3 and adult worm antigens were all significantly higher (P < 0.01 or P < 0.05) in LFEC lambs than in HFEC lambs. PMID- 8720564 TI - Prevalence of dicrocoeliosis in sheep and goats in Himachal Pradesh, India. AB - A survey incorporating field and abattoir studies was carried out on the prevalence of Dicrocoelium dendriticum in sheep and goats in the Kangra valley of Himachal Pradesh (India). Coprological studies on these migratory animals revealed that 8.1% of sheep and 4.1% of goats were positive for dicrocoeliosis, with a mean number of eggs per gram of faeces (EPG) of 702.1 for sheep and 566.7 for goats. The necropsy liver examination of slaughtered animals revealed that 24.1% of sheep and 12.3% of goats were infected with D. dendriticum and they had mean fluke burdens of 247.2 (sheep) and 298.1 (goats). No significant differences in fluke burden, liver lesion score and EPG count were observed between sheep and goats. The seasonal distribution of dicrocoeliosis indicated a higher percentage of infection in autumn and winter as compared with spring and summer. The potential role of migratory sheep and goats in the epizootiology of dicrocoeliosis is discussed. PMID- 8720565 TI - Dietary protein influences upon immunity to Nematodirus battus infection in lambs. AB - Several indices of the immune response to Nematodirus battus infection in lambs offered differing levels of dietary protein were quantified. Lambs were offered either a complete basal ruminant diet (13.2% crude protein (CP)) or the same diet supplemented with fish meal as a source of rumen bypass protein (18.3% CP). Lambs from each dietary treatment group were given either a 7-week continuous trickle infection with N. battus L3 or remained uninfected. All lambs were drenched with anthelmintic at week 8 post-infection (PI), challenged with a single dose of 30,000 N. battus L3 1 week later, and killed 9 days post-challenge (PC). Previous infection induced a significant reduction in worm burdens (p < 0.001) and enhancement of immune responses when compared to challenge controls. Among previously infected lambs, protein supplementation did not reduce worm burdens significantly, although there was a trend for fewer worms in the supplemented lambs. However, a significant increase in mucosal globule leucocyte (p < 0.05) and eosinophil (p < 0.05) numbers was evident. Supplementation (p < 0.05) and previous infection (p < 0.001) both enhanced serum anti-worm IgG titres over time. Peripheral blood eosinophil counts were not affected by supplementation but were significantly elevated over time as a result of previous infection (p < 0.001). Since there were no significant differences in worm burdens of supplemented and unsupplemented previously infected lambs, it was of interest to determine whether lambs possessed an innate ability to regulate their parasite burden. Hence they were re-grouped based on an arbitrary cut-off burden of 1000 worms. High responders (HR) had burdens below 1000 worms, while low responders (LR) had burdens above this value and challenge controls were pooled. The data were re-analyzed based on these groupings and showed significant reduction in worm burdens between all three groups (p < 0.001). Globule leucocytes were the only cell type that appeared to be significantly more abundant in the intestinal mucosa of HR (p < 0.001). Serum antibody responses (p < 0.05) and peripheral blood eosinophil counts (p < 0.01) were significantly elevated over time in accord with the degree of responsiveness. The results of this study suggest that supplementation of protein upon an adequate basal diet of lambs previously exposed to N. battus does not significantly enhance worm regulation despite significant increases in cellular and antibody responses. The immunity acquired is characterized by reduction in worm burdens, elevated anti-worm antibodies and a cellular inflammatory response. The identification of HR and LR essentially shows that when the protein supply is adequate, the predominant host effect influencing the pathogenicity of the parasites is the level of genetically determined susceptibility of the host. PMID- 8720566 TI - Isotype-specific antibody responses to the surface-exposed antigens of adult and larval stages of Dictyocaulus viviparus in infected and vaccinated calves. AB - The antibody responses to the surface-exposed antigens of living larval and adult Dictyocaulus viviparus were measured by quantitative immunofluorescence using sera from calves infected with, or vaccinated against, the parasite. In infected animals, the surface of the sheath of the third-stage larvae (L3) (retained cuticle of second-stage larvae (L2)) proved highly immunogenic despite the fact that it is thought to be shed prior to parasite penetration of the host intestine. When responses to the surface of exsheathed larvae (L3 cuticle) were measured, a high level of heterophile IgM antibody was detected in the serum of animals that had not been previously exposed to the parasite and, following infection, a specific IgG response was detected against the exsheathed L3 surface. The antibody response, however, was less marked than that observed against the intact L3 sheath. Responses of patently infected animals to the adult surface showed an initial IgM response that was superseded with time by IgG1 and IgG2 responses. Vaccinated animals showed only low level responses to the surfaces of the L3 sheath, L3 cuticle and adult stages following immunisation with two doses of irradiated larvae. The immunised animals produced a strong antibody response to the larval surface antigens following challenge with infective larvae but they failed to produce antibody to the surface of adult parasites. These results show that the surfaces of all the stages of D. viviparus examined are immunogenic in infected calves and, depending on the developmental stage, infection regime, or time of infection, high levels of parasite-specific IgG1 or IgM are stimulated. It has previously been shown that significant levels of protective immunity can be obtained in naive animals following passive transfer of serum from infected calves. Thus, the antibody responses detected in the work reported here may be of relevance in protective immunity against dictyocaulosis. PMID- 8720567 TI - Long-term subclinical infection with Trichostrongylus colubriformis: effects on food intake, diet selection and performance of growing lambs. AB - Growing lambs (mean liveweight 22.4 kg) were used to test for the effects of long term subclinical infection with Trichostrongylus colubriformis (2500 larvae per day) on the voluntary food intake (VFI), diet selection and performance of sheep given a choice between two foods of different protein content (low versus high). Sheep were either uninfected controls or parasitised for 27 weeks (Group 27P). Additional sheep were infected for either 9 or 18 weeks (before and after the development of complete resistance to parasites, as assessed by faecal egg counts) and then treated with anthelmintic to remove the parasites (Groups 9P and 18P). Group 18P remained uninfected after anthelmintic treatment whereas Group 9P was infected again for the last 9 weeks after a 9 week worm-free period. The VFI declined or remained static from Week 5 up to Week 11/12 of continuous infection, for the 27P and 18P groups. For 9P sheep, VFI recovered within a few days after the anthelmintic administration and was unaffected during re-infection over the last 9 weeks of the experiment. Diet selection changed in such a pattern that the proportion of the high protein food (prop H) in the selected diet started to increase from Week 6 of the infection. The prop H selected remained higher over the interval 10-18 weeks of the continuous infection for 27P and 18P groups than the uninfected controls. There were no significant differences in diet selection among any of the groups beyond Week 18 of the experiment. The infection caused a growth check from Week 7 to approximately Week 12/14 of dosing; these was no evidence of compensatory growth following the development of resistance or recovery. The results are consistent with the view that sheep given a choice between two appropriate foods will modify their diet selection in order to moderate the adverse effects of subclinical nematode infection. The performance and diet selection of all sheep beyond Week 18 of infection suggest that no obvious nutritional penalties are incurred with the continuation of the infection once the animals have become resistant to parasites. PMID- 8720568 TI - Ocular larva migrans and histopathological lesions in mice experimentally infected with Baylisascaris transfuga embryonated eggs. AB - The ability of Baylisascaris transfuga larvae to cause ocular larva migrans (OLM) in mice was examined. Mice were given approximately 3500 infective eggs of B. transfuga per os. Their eyes were removed and examined either microscopically or histologically at various intervals post-infection. Larvae were recovered beginning 7 days after infection. Histologically, free larvae were observed in the posterior chamber and within the ocular membranes. Larval granulomas were present in the choroid with involvement of retinal layers. It was concluded that B. transfuga larvae have the ability to produce OLM in mice following oral infection and should be considered as possible agents of OLM in other animals and in human beings. PMID- 8720569 TI - Coccidial and helminth infections in goats kept indoors in the Netherlands. AB - An investigation was carried out on coccidial and helminth infections in goats kept indoors on five farms in the Netherlands. The goats were individually sampled. Coccidial oocysts were identified and nematode eggs counted. Larval cultures were made and infective larvae identified to the generic or species level. The goats were divided into three groups according to their age: kids, those weaned but not served, and older goats. Oocysts were found in 26 out of 27 kids (96.3%), in 52 out of 55 weaned but not served goats (94.5%), and in 72 out of 110 older goats (65.5%). Eimeria ninakohlyakimovae was the most common species in all groups with an overall prevalence of 82%, followed by Eimeria arloingi (78%) and Eimeria aspheronica (53.3%). Other species present were Eimeria alijevi (50%), Eimeria christenseni (39.3%), Eimeria caprina (38.7%), Eimeria hirci (28.7%) and Eimeria jolchijevi (7.3%). Nematode eggs were seen on four of the five farms. It appeared that on two farms some goats, although now kept indoors, had been on pasture in the past. These goats excreted low numbers of eggs of Ostertagia and/or Trichostrongylus, Strongyloides papillosus, Haemonchus contortus, Trichuris spp. and larvae of Muellerius capillaris. In goats that were kept exclusively indoors, infections with Strongyloides papillosus were found on two farms in the older group and on one farm also in the group weaned but not served. On two farms Skrjabinema ovis was incidentally observed by the farmer in older goats. This was confirmed by faecal examinations on one farm. All kids were free of nematode infection. PMID- 8720570 TI - Apyrase activity and adenosine diphosphate induced platelet aggregation inhibition by the salivary gland proteins of Culicoides variipennis, the North American vector of bluetongue viruses. AB - Salivary gland homogenates of Culicoides variipennis, the primary vector of bluetongue (BLU) viruses in North America, were analyzed for apyrase activity. Apyrase (ATP diphosphohydrolase, EC 3.6.1.5) is an anti-hemostatic and anti inflammatory salivary enzyme of most hematophagous arthropods. The enzyme activity was measured by the release of orthophosphate using ATP, ADP, and AMP as substrates with Ca2+ as the divalent cation. ATPase (11.5 +/- 1 mU/pair of glands), ADPase (7.3 +/- 0.7 mU/pair of glands), and insignificant (P < 0.05) AMPase (0.07 mU/pair of glands) activities were detected in female salivary glands. Male salivary glands contained lower amounts of ATPase and ADPase activity (P < 0.05). The ATPase and ADPase activities were greatest at pH 8.5, and were similarly activated by Mg2+. Molecular sieving HPLC of salivary gland homogenates generated a single peak which coincided with ATPase and ADPase, but no AMPase, activity; the protein has an estimated molecular mass of 35,000 Da. ATPase and ADPase activity, and total protein concentration, were reduced (P < 0.05) in the salivary glands of females after taking a blood meal from a sheep. Salivary gland homogenates also inhibited ADP-induced platelet aggregation in vitro. It is concluded that the salivary ATPase and ADPase activities of C. variipennis reside in one enzyme, and that this enzyme is likely an apyrase. The apyrase activity is thought to be responsible for the inhibition of ADP-induced platelet aggregation, as indicated by the apparent discharge of apyrase from salivary glands into the host during blood feeding. This suggests that apyrase is one of the salivary proteins present in C. variipennis acting as antigens in the development of Culicoides hypersensitivity in ruminants and horses. Apyrase may inhibit an inflammatory response at the feeding site through the subsequent degradation of its end-product, AMP, to adenosine, a potent anti-inflammatory substance, by the ecto-5' nucleotidase activity of neutrophils. PMID- 8720571 TI - The sterilizing effect of pour-on flumethrin on the camel tick, Hyalomma dromedarii (Acari: Ixodidae). AB - The relationship between the efficacy of 1% flumethrin in pour-on formulation on the fertility of engorged female Hyalomma dromedarii and exposure time was investigated. Using a contact method, ticks were exposed in vitro to 87 micrograms active ingredient for 1, 5, 10, 30 or 720 min. Fertility inhibition was related to the exposure time. No significant effect was obtained after 1 min exposure, but 5 or 10 min exposure reduced the percentage of females ovipositing, egg mass weight, number of eggs, percentage of females laying eggs that hatch and the conversion efficiency of female weight to egg mass weight. The action of flumethrin was highly pronounced after 30 min; out of ten females, only one oviposited on the 19th day after exposure and the eggs did not hatch. Females exposed for 12 h were paralysed and showed complete sterilization. The pre oviposition and egg incubation periods of females exposed for 1, 5 or 10 min were prolonged due to the action of the drug. PMID- 8720572 TI - Viability after thawing and dilution of simultaneously cryopreserved vaccinal Babesia bovis and Babesia bigemina strains cultured in vitro. AB - A live, frozen experimental vaccine containing Babesia bovis and Babesia bigemina multiplied in vitro was stored in liquid nitrogen after simultaneous cryopreservation using glycerol as cryoprotectant. The viability of the vaccine was tested by inoculating (subcutaneously) three groups of seven steers each, 2, 12 and 24 h after thawing at 40 degrees C and dilution to obtain a dose of 2 x 10(7) of each organism. All vaccinated cattle developed detectable parasitaemia in thin and/or thick blood smears. No statistically significant differences in the prepatent period were detected amongst the cattle groups (analysis of variance). This prepatent period was 12.3 days for B. bovis and 8.4 days for B. bigemina. Vaccinal organisms derived from in vitro culture systems may replace antigens obtained from in vivo culture to produce vaccine against cattle babesiosis. PMID- 8720573 TI - Trypanocidal value of liposomal diminazene in experimental Trypanosoma brucei evansi infection in mice. AB - The trypanocidal value of liposomal diminazene was examined. Three hundred mice were divided randomly into six groups of 50 mice each. Groups 1, 2 and 3 were pretreated 2 h before Trypanosoma brucei evansi challenge with 0.2 ml saline solution, 0.2 ml diminazene solution (5 mg ml-1) and 0.2 ml liposomal solution (5 mg diminazene per ml), respectively. Mice in Group 1 all died within 6 days, 33 in Group 2 and four in Group 3 died within 15 days after challenge, respectively. Groups 4, 5, and 6 were treated 4 days after T. b. evansi challenge with 0.2 ml saline solution 0.2 ml diminazene solution (5 mg ml-1) and 0.2 ml liposomal solution (5 mg diminazene per ml), respectively. Mice in Group 4 all died within 6 days, 19 in Group 5 and two in group 6 died within 15 days after challenge, respectively. The remaining mice survived for more than 30 days, were symptom free and behaved normally. No adverse effects associated with treatment were noted. PMID- 8720574 TI - Helminth infections of sheep in North Sumatra, Indonesia. AB - Gastrointestinal tracts of 73 indigenous sheep were obtained from the Medan abattoir in North Sumatra, Indonesia, and examined for the presence of helminths. A total of 13 species of helminths, eight nematodes, one cestode and four trematodes were encountered. All sheep were infected by more than one species of nematodes. Trichostrongylus colubriformis, T. axei and Haemonchus contortus were the most common species. The intensity of the nematode infections was very high: the average burden was over 7500 worms. Trichostrongylus spp. accounted, on average, for 81.5% of the total nematode burden. No influence of age on total nematode burden could be shown. Eurytrema pancreaticum and Schistosoma spindale were found in 23.3% and 4.1% of the examined sheep, respectively. The results are discussed in relation to common management and helminth control systems. PMID- 8720575 TI - Direct evidence for a role of glutamate in the expression of the opioid withdrawal syndrome. AB - To investigate the role of glutamate in the expression of the withdrawal signs from opioids, rats were intracerebroventricularly (i.c.v.) infused continuously with morphine (a mu-opioid receptor agonist, 26 nmol/microliters per h) or butorphanol (a mixed mu/delta/kappa-opioid receptor agonist, 26 nmol/microliters per h) through osmotic minipumps for 3 days. An i.c.v. injection of glutamate (5 and 50 nmol/5 microliters) dose dependently induced withdrawal signs in morphine- or butorphanol-dependent animals. The withdrawal signs precipitated by the glutamate injection were comparable to those precipitated by an opioid receptor antagonist, naloxone (48 nmol/5 microliters), except for the expression of some specific behaviors and the duration of withdrawal signs. Glutamate or naloxone challenge failed to precipitate any withdrawal signs in saline controlled animals. On the other hand, the expression of the withdrawal signs precipitated by glutamate or naloxone in opioid-dependent animals was completely blocked by pretreatment with MK-801 [a NMDA receptor antagonist, (+)-5-methyl-10,11-dihydro 5H-dibenzo[a,d]cycloheptan-5,10-imine], 0.1 mg/kg, i.p. These unique actions of glutamate in continuously opioid-infused rats suggest that a rapid central release of glutamate may be a key factor in the expression of withdrawal signs from opioids. Furthermore, this effect may be mediated by the NMDA subtype of glutamate receptors. PMID- 8720576 TI - Nitric oxide modulates retention of immobility in the forced swimming test in rats. AB - Although originally developed as a possible screen for antidepressants, the Porsolt forced swimming test has more recently been extensively used as a model for studying the involvement of the endocrine system in the acquisition and retention of behavioural responses. In previous studies we have shown that while adrenalectomised rats acquire the immobile response normally, they are unable to retain it on retest next day. In the present study we show that retention of the immobile response in the Porsolt swim test is impaired in intact rats given the nitric oxide (NO) inhibitor L-N-arginine methyl ester (L-NAME), in a dose- and time-dependent manner. At a dose of 50 mg/kg levels of immobility are similar to those in adrenalectomised animals, an effect reversed by the simultaneous administration of L-arginine (50 mg/kg). L-Arginine also reverses the behavioural effect of adrenalectomy, and L-NAME blocks the ability of dexamethasone or the kappa-selective opioid ketocyclazocine to reverse the effect of adrenalectomy on retention of the immobile response. We conclude that the kappa-opioid and glucocorticoid mediated pathways previously shown to independently facilitate retention are mediated by nitric oxide. PMID- 8720577 TI - Morphine-induced catalepsy is augmented by NMDA receptor antagonists, but is partially attenuated by an AMPA receptor antagonist. AB - High doses of morphine produce a state of behavioural inactivity and muscular rigidity. This type of 'catalepsy' is clearly different from the state which is produced by the administration of neuroleptics, e.g. haloperidol. While haloperidol-induced catalepsy can easily be antagonised by NMDA receptor antagonists, there has been a report that the non-competitive NMDA receptor antagonist (+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d] cyclohepten-5,10-imine (MK 801) potentiates morphine-induced catalepsy. The aim of this study was to further examine the role of glutamate receptors in the mediation of morphine-induced catalepsy. To this end we coadministered morphine (20, 40, 60 mg/kg i.p.) with MK 801 (0.1 and 0.3 mg/kg i.p.), the competitive NMDA receptor antagonist DL-(E)-2 amino-4-methyl-5-phosphono-3-pentoic acid (CGP 37849) (2 and 6 mg/kg i.p.), or 1 (4-aminophenyl)-4-methyl-7,8-methylen-dioxy-5H-2,3- benzodiazepine (GYKI 52466) (2 and 4 mg/kg), an antagonist of the AMPA type of glutamate receptors, respectively. The degree of catalepsy was assessed using two different methods, the 'bar/podium/grid' test which is commonly used to measure neuroleptic-induced catalepsy, and a test for the presence or absence of righting reflexes after turning the animals into a supine position. It was found that in the 'bar/podium/grid' test coadministration of both NMDA receptor antagonists significantly and dose-dependently augmented morphine-induced catalepsy. The results using the AMPA receptor antagonist were less clear since the lower dose of GYKI 52466 tended to attenuate the morphine effect whereas the higher dose augmented morphine-induced catalepsy in some cases. While placing the animals on the bar and on the podium produced essentially the same results, the grid was found to be inapplicable for the measurement of morphine-induced catalepsy since the animals did not cling to the grid and fell off almost immediately after being released from the experimenter's hand. With respect to the righting reflexes it was found that the number of animals not showing these responses increased when MK-801 or CGP 37849 was coadministered with morphine. In contrast, most of the animals treated with GYKI 52466 and morphine displayed intact righting reflexes. It is concluded that glutamatergic transmission plays an important role in the mediation of morphine-induced catalepsy, though different to that of haloperidol induced catalepsy, and that NMDA and AMPA receptors are differentially involved in different aspects of the associated behavioural state. PMID- 8720578 TI - Selective adenosine A2A receptor/dopamine D2 receptor interactions in animal models of schizophrenia. AB - In the apomorphine-induced climbing mouse assay, the potencies of the selective adenosine A1 receptor agonist, N6-cyclohexyladenosine (CHA), and the selective A2A adenosine receptor agonist, 2-p-(2-carboxyethyl) phenethylamino 5'-N-ethyl carboxamidoadenosine (CGS 21680), and various dopamine receptor antagonists were as follows: SCH 23390 = haloperidol > raclopride > CHA = CGS 21680. While in catalepsy, their potencies were SCH 23390 > haloperidol > raclopride > CGS 21680. CHA failed to induce catalepsy due to significant sedation/ataxia. The combined administration of the ED15 dose of CHA failed to potentiate the ED50 value of SCH 23390, raclopride, or haloperidol in the apomorphine-induced climbing mouse assay. However, the combined administration of the ED15 dose of CGS 21680 significantly decreased the ED50 of raclopride by 8.0-fold and haloperidol by 35 fold. The adenosine A2A receptor antagonist, 1,3,7-trimethyl-8-(3 chlorostyryl)xanthine (CSC), significantly decreased catalepsy induced by raclopride and haloperidol, while the adenosine A1 receptor antagonist, 1,3 dimethyl-8-phenylxanthine (8-PT), was ineffective. The present results show that in behavioral assays predictive for antipsychotic activity, adenosine receptor agonists block behaviors in a similar manner to dopamine receptor antagonists. PMID- 8720579 TI - Evidence for specific involvement of 5-HT1A and 5-HT2A/C receptors in the expression of patterns of spontaneous motor activity of the rat. AB - The 5-HT1A and the 5-HT2A/C receptor agonists 8-hydroxy-2-(di-n propylamino)tetralin (8-OH-DPAT) (0.006-0.4 mg kg-1 s.c.) and (+/-)-1-(2,5 dimethoxy-4-iodophenyl)-2-aminopropane (DOI) (0.05-4.0 mg kg-1 s.c.), respectively, produced a similar stereotyped forward locomotion in rats, although the intensity of the behavioral change was considerably less with DOI. The stereotyped forward locomotion was accompanied by a slight decrease in total activity, suppression of rearing behavior and an increased activity in the periphery of the open-field arena. In support of receptor specificity, the effects of 8-OH-DPAT and DOI could be antagonised by pretreatment with the 5 HT1A/B and the 5-HT2A/C receptor antagonists (-)-pindolol (2 mg kg-1 s.c.) and ritanserin (2 mg kg-1 s.c.), respectively. In addition, (-)-pindolol, but not the selective beta-adrenoceptor antagonist betaxolol, markedly enhanced the behavioral effects produced by DOI. The nature of these specific actions and interactions in terms of pre- and post-synaptic serotonergic mechanisms remains an important question. PMID- 8720580 TI - Effects of cannabinoid receptor stimulation and blockade on catalepsy produced by dopamine receptor antagonists. AB - The ability of cannabinoid receptor stimulation or blockade to alter catalepsy produced by dopamine D1 and D2 receptor antagonists was studied in rats. The cannabinoid receptor antagonist SR 141716A (N-(piperidin-1-yl)-5-(4-chlorophenyl) 1-(2,4-dichlorophenyl)-4-me thyl-1H- pyrazole-3-carboxamidehydrochloride) (0.5 and 2.5 mg/kg) reduced catalepsy elicited by the cannabinoid receptor agonist CP 55,940 (1 alpha,2-(R)-5-(1,1-dimethylheptyl)-2-[5-hydroxy-2-(3-hydroxypropyl ) cyclohexyl-phenol) (0.5 mg/kg). However, SR 141716A (0.5 and 2.5 mg/kg) did not decrease catalepsy produced by the dopamine D1 receptor antagonist SCH 23390 (R (+)-7chloro-8-hydroxy-3-methyl-1-phenyl-2,3,4,5- tetrahydro-1-H-3-benzazepine) (0.5 mg/kg) or the dopamine D2 receptor antagonist raclopride (S(-)-3,5-dichloro N-(1-ethyl-2-pyrrolidinyl)-methyl-6-methoxysalicylami de) (2.5 mg/kg), suggesting that, under these conditions, endogenous cannabinoid ligands do not modulate the cataleptic effects of dopamine D1 or D2 receptor antagonists. In contrast, CP 55,940 (0.025 and 0.1 mg/kg), at doses which do not produce catalepsy when administered alone, enhanced catalepsy produced by SCH 23390 and raclopride. These results suggest that stimulation, but not blockade, of brain cannabinoid receptors modifies catalepsy behavior produced by selective dopamine D1 and D2 receptor blockade. PMID- 8720581 TI - An autoradiographic study of [3H]flunitrazepam binding sites in the brain of rat made tolerant to and dependent on pentobarbital. AB - The effects of continuous administration of pentobarbital on the benzodiazepine receptor labeled by [3H]flunitrazepam were investigated. Animals were made tolerant to pentobarbital by intracerebroventricular (i.c.v.) infusion with pentobarbital (300 micrograms/10 microliters/h) for 6 days through pre-implanted canulae connected to osmotic mini-pumps. The dependent rats were assessed 24 h after cessation of pentobarbital infusion. Changes in [3H]flunitrazepam binding were examined in 37 brain regions at a concentration of [3H]flunitrazepam of 1 nM. In subsequent saturation studies, the binding parameters Bmax and KD were also investigated in 17 brain regions, most of which showed significant changes in [3H]flunitrazepam binding in experiments using a fixed concentration of radioligand. The pentobarbital-tolerant rats showed a significant increase in Bmax with an increase in KD for [3H]flunitrazepam in the ventroposterior nucleus of thalamus. In the dependent rats, a significant increase in Bmax for [3H]flunitrazepam binding, without a change in KD, was observed in all layers of the frontal cortex, the caudate-putamen, olfactory tubercle, and some nuclei in thalamus, compared to those in the control. Increased [3H]flunitrazepam binding in the molecular layer of the olfactory bulb, the ventral pallidum, and the cerebellum of the pentobarbital dependent rats at a fixed concentration of [3H]flunitrazepam was also observed. There was no significant change in [3H]flunitrazepam binding in the hippocampus and several nuclei of the brain stem. These findings suggest that benzodiazepine receptors are closely involved in the development of tolerance to and dependence on pentobarbital. Further studies on changes in gamma-aminobutyric acid (GABA)A receptor subunit mRNA or the effects of pentobarbital on GABAA receptor phosphorylation would be necessary for an explanation of the precise mechanisms underlying the development of tolerance to and dependence on pentobarbital. PMID- 8720583 TI - Effects of acute and repeated administration of citalopram on extracellular levels of serotonin in rat brain. AB - The effects of acute (2 days) and repeated (21 days) administration (50 mg/kg in the diet) of the selective serotonin (5-HT, 5-hydroxytryptamine) reuptake inhibitor, citalopram, on extracellular levels of 5-HT and their modulation by terminal autoreceptors in the hypothalamus of freely moving rats were compared in vivo by microdialysis. When studied without washout, extracellular levels of 5-HT were increased by both acute and repeated citalopram administration. In rats treated repeatedly, extracellular 5-HT levels were 43% (but not significantly) greater than in those treated acutely. Extracellular levels of 5-HT in control and citalopram-treated rats were similar when measured after 24 h washout. The enhancing effect of non-selective serotonergic autoreceptor antagonists, methiothepin (100 microM) or 1-(1-naphthyl)piperazine (NP) (10 microM), administered through the microdialysis probe, after 24 h washout, was similar in both control and chronically treated groups. These results suggest that repeated administration of citalopram followed by a washout of 24 h does not lead to desensitization of the terminal autoreceptor as measured in vivo in contrast to the effects we have shown previously in vitro. In rats treated chronically with citalopram without washout, methiothepin had a greater maximal effect on 5-HT outflow in comparison to rats receiving acute citalopram treatment. This finding suggests that a 5-HT autoreceptor antagonist or a combination of such a drug with a 5-HT uptake inhibitor would produce a greater increase of extracellular levels of 5-HT in hyposerotonergic states such as depression. PMID- 8720582 TI - Effects of K+ channel blockers and openers on antinociception induced by agonists of 5-HT1A receptors. AB - The modulation by K+ channel-acting drugs of the antinociceptive effect of several 5-HT1A receptor agonists was examined with the hot plate test in mice. All the 5-HT1A receptor agonists tested induced dose-dependent antinociception, the order of potency being (+/-)-8-hydroxy-2-(di-n-propyl-amino)tetralin (8-OH DPAT) > buspirone > or = lesopitron > or = tandospirone. The blockers of ATP sensitive K+ channels (KATP) gliquidone and glipizide (1-4 and 16-64 micrograms/mouse i.c.v., respectively) reduced the antinociceptive effect of 8-OH DPAT, whereas cromakalim (32-64 micrograms/mouse i.c.v.), an opener of KATP channels, enhanced the effect. In contrast, 4-aminopyridine (25-250 ng/mouse i.c.v.) and tetraethylammonium (10-20 micrograms/mouse i.c.v.), which antagonize several non-ATP-dependent K+ conductances, were inactive. The same results were found with other agonists of 5-HT1A receptors (lesopitron, buspirone and tandospirone): gliquidone inhibited whereas cromakalim increased their antinociceptive effects. None of the K+ channel-acting drugs modified the binding of [3H]8-OH-DPAT to hippocampal membranes, whereas all the 5-HT1A receptor agonists displaced the ligand. These results suggest that ATP-sensitive K+ conductances are involved in the antinociception induced by agonists of 5-HT1A receptors. PMID- 8720584 TI - The acid metabolite of ZD7114 is a partial agonist of lipolysis mediated by the rat beta 3-adrenoceptor. AB - Experiments were performed to characterise the lipolytic effects of the acid metabolite, ZM215001, ((S)-4-[2-hydroxy-3-phenoxy-propylamino-ethoxy] phenoxyacetic acid) of the putative beta 3-adrenoceptor agonist, ZD7114 ((S)-4-[2 hydroxy-3-phenoxy-propylamino-ethoxy]-N-(2-methoxyethyl) phenoxyacetamide) on isolated rat white adipocytes. ZM215001 was used for these studies since it is the predominant moiety after in vivo administration of ZD7114. The agonist properties of ZM215001 were assessed in comparison to the standard nonselective beta-adrenoceptor agonist (+/-)-isoprenaline and the beta 3-adrenoceptor selective agonist BRL 37344. Isoprenaline, BRL 37344 and ZM215001 all stimulated the rate of free fatty acid release from isolated adipocytes with the order of potency being BRL > isoprenaline > ZM215001. The maximum effect of BRL 37344 was equivalent to that of isoprenaline, but ZM215001 achieved only 30% of the maximum isoprenaline response. ZM215001 competitively antagonised the lipolytic response to BRL 37344 (pA2 = 7.26), whereas the agonist effects of BRL 37344 were not antagonised competitively by the selective antagonists ICI 118551 and CGP 20712A, at concentrations which would be expected to block beta 1- and beta 2 adrenoceptors respectively. These results indicate that ZM215001 has low intrinsic activity at the rat adipocyte beta 3-adrenoceptor, and is a partial agonist of lipolysis in rat white adipocytes. PMID- 8720585 TI - The nicotinic receptor agonists (-)-nicotine and isoarecolone differ in their effects on dopamine release in the nucleus accumbens. AB - This study compared the effect of the nicotinic receptor agonists, (-)-nicotine and isoarecolone, on the mesolimbic dopamine system of the rat using in vivo microdialysis. Previous studies showed that (-)-nicotine but not isoarecolone produced a locomotor activating effect, and that this was probably mediated by increased concentrations of dopamine in the nucleus accumbens. Nicotine (0.4 mg/kg s.c.) significantly increased extracellular concentrations of dopamine and of dihydroxyphenylacetic acid (DOPAC) by 75-80% in nucleus accumbens of rats. Isoarecolone (3.2-32 mg/kg s.c.) had no significant effect on either dopamine or DOPAC levels in this brain region and neither drug affected extracellular levels of 5-hydroxy indole acetic acid. Both nicotine and isoarecolone induced head bobbing behaviour. Pretreatment with ketanserin reduced nicotine-induced head bobbing suggesting a serotonergic mechanism. In conclusion, the absence of locomotor activation after administration of isoarecolone may be related to its failure to activate the mesolimbic dopamine system. PMID- 8720586 TI - Effect of (+/-)-8-hydroxy-2-(di-n-propylamino)tetralin hydrobromide on spinal motor systems in anesthetized intact and spinalized rats. AB - In the present study, we examined the effect of the 5-HT1A receptor agonist (+/-) 8-hydroxy-2-(di-n-propylamino)tetralin hydrobromide (8-OH-DPAT), on the mono- and polysynaptic reflexes in intact and spinalized rats. 8-OH-DPAT (10 micrograms/kg i.v.) significantly potentiated the amplitude of the monosynaptic reflex in intact rats. In contrast, 8-OH-DPAT (30 and 100 micrograms/kg i.v.) produced a significant dose-related inhibition of the amplitude of the monosynaptic reflex in spinalized rats. These results suggest that 8-OH-DPAT predominantly excites spinal motor systems at the supraspinal site, and inhibits such systems at a spinal cord site. PMID- 8720587 TI - Inhibition of nitric oxide formation with L-canavanine attenuates endotoxin induced vascular hyporeactivity in the rat. AB - L-Canavanine, a selective inhibitor of inducible nitric oxide (NO) synthase, has beneficial effects on the circulatory failure of rats with endotoxin shock. To investigate the direct relationship between these beneficial effects and the inhibition of the formation of NO in response to L-canavanine in endotoxin shock in the rat, we detected changes in venous nitrosyl-hemoglobin (NO-hemoglobin) levels using an electron spin resonance (ESR) assay. Anaesthetized rats were injected with lipopolysaccharide (10 mg/kg i.v.). 1 h after the lipopolysaccharide injection, the rats were divided into four groups: a lipopolysaccharide group receiving 0.3 ml of saline hourly, an L-canavanine 10 or an L-canavanine 20 group receiving L-canavanine 10 or 20 mg/kg i.v. hourly, respectively, and an L-NAME group receiving NG-nitro-L-arginine methyl ester (L NAME) 15 mg/kg followed by 10 mg/kg i.v. hourly. A sham group received saline instead of lipopolysaccharide, and an L-canavanine group received L-canavanine 20 mg/kg i.v. hourly, 1 h after the saline injection. At 5 h after the lipopolysaccharide or saline injection, pressor responses to noradrenaline (1 microgram/kg i.v.) were obtained. In the lipopolysaccharide group, lipopolysaccharide caused a progressive decrease in mean arterial pressure and an impairment of pressor responsiveness to noradrenaline. Administration of L canavanine or L-NAME attenuated the endotoxin-induced hypotension and vascular hyporeactivity to noradrenaline. L-Canavanine did not alter mean arterial pressure and the pressor response to noradrenaline in the L-canavanine group. The endotoxin-induced increases in venous levels of NO-hemoglobin were significantly inhibited by L-canavanine or L-NAME. These data indicate that the beneficial hemodynamic effects of L-canavanine are associated with inhibition of the enhanced formation of NO by inducible NO synthase in a rat model of endotoxin shock. L-Canavanine is a potential agent in the treatment of endotoxin shock. PMID- 8720588 TI - Tranilast suppresses intimal hyperplasia after photochemically induced endothelial injury in the rat. AB - Intimal thickening in the femoral artery of spontaneously hypertensive rats (SHR) was initiated by endothelial damage induced by the photochemical reaction between green light and systemic rose bengal. This model represents a non-mechanical method of producing vessel wall denudation. Neointima formation was assessed by calculating the cross-sectional area of intima, media and lumen, using computer analysis. Tranilast (30, 100 and 300 mg/kg, p.o.), administered 2 days prior to endothelial injury, reduced intimal area by 29, 62 and 87%, respectively, compared to that of vehicle-treated controls. In cultured SHR-derived vascular smooth muscle cells, tranilast produced concentration-dependent inhibition of mitogenesis, whether stimulated by platelet-derived growth factor, basic fibroblast growth factor, insulin-like growth factor or fetal bovine serum. These results suggest that tranilast may be effective in preventing coronary restenosis. PMID- 8720589 TI - Semotiadil improves survival of rats with monocrotaline-induced pulmonary hypertension: comparison with diltiazem. AB - We compared the effects of semotiadil, a novel Ca2+ channel blocker, with those of diltiazem on survival and regression of right ventricular hypertrophy and media thickening of pulmonary arteries in a rat model of pulmonary hypertension. Pulmonary hypertension was induced by a single injection of monocrotaline (80 mg/kg). Four weeks later, after pulmonary hypertension was confirmed, oral administration of semotiadil (10, 30, or 100 mg/kg/day) or diltiazem (100 or 300 mg/kg/day) was initiated. The rats were observed for 3 weeks. Survival was significantly longer in the group that received semotiadil 100 mg/kg/day than in the groups treated with diltiazem 100 or 300 mg/kg/day. Media thickness and smooth muscle area in pulmonary arteries were significantly less in rats treated with semotiadil 100 mg/kg/day than in animals treated with diltiazem 100 mg/kg/day. The right ventricle to left ventricle mass ratio, right ventricular wall thickness, and right ventricular myocardial fiber diameter were equal in these two groups. Semotiadil 100 mg/kg/day improved the survival of rats, which responded with a significant regression of right ventricular hypertrophy and media thickening of pulmonary arteries in comparison with rats treated with diltiazem 100 or 300 mg/kg/day. PMID- 8720590 TI - The specific effect of Mn2+ on the tonic components of receptor-mediated contractions in isolated vas deferens of the guinea pig. AB - Intracellular accumulation of Mn2+ augmented the contractions induced by norepinephrine and acetylcholine in the guinea pig isolated vas deferens. Contractions repeatedly induced by norepinephrine, acetylcholine, or a high concentration of K+ decreased depending on the incubation time in Ca(2+)-free medium. The rate of decrease of all contractions was delayed by intracellularly accumulated manganese. In the Mn(2+)-loaded preparations, the tonic components of the contractions induced by norepinephrine and acetylcholine, but not K+, were highly resistant to extracellular Ca2+ elimination. Ryanodine abolished the initial phasic component but did not affect the tonic component of norepinephrine and acetylcholine-contractions in Mn(2+)-loaded preparations in Ca(2+)-free medium. In Ca(2+)-depleted preparations, the tonic contraction induced by norepinephrine was restored after the Mn(2+)-loading procedure, and the magnitude of this tonic contraction was comparable to the tonic component of the norepinephrine contraction in the normal medium before Mn2+ loading. The tonic contraction was reproducible in medium without either Mn2+ or Ca2+. These results suggested that intracellular Mn2+ can support norepinephrine-induced tonic contractions. In the Ca(2+)-depleted Mn(2+)-loaded preparations, K+ also induced a tonic contraction in the presence of extracellular Mn2+. However, this contraction was much smaller than that induced by norepinephrine. These results suggested that intracellular Mn2+ augmented contractions not only via an increase in intracellular Ca2+ availability but also via the direct action of Mn2+ on contractile mechanisms, and that this action is highly specific for developing and/or maintaining tonic contractions mediated by receptor activation in the guinea pig isolated vas deferens. PMID- 8720591 TI - Some studies on the effects of alpha-chymotrypsin on mast cells from the rat and other species. AB - We have examined the effect of alpha-chymotrypsin on isolated mast cells from different sources. The enzyme induced a dose-dependent secretion of histamine from purified and non-purified populations of rat peritoneal mast cells. The release was non-cytotoxic and was inhibited by metabolic blockers and extremes of temperature. The process was relatively slow, being essentially complete within 20 min, and was unaffected by phosphatidylserine. A substantial component of the secretion persisted in the absence of extracellular Ca2+. The release was suppressed by extremes of pH and a variety of anti-allergic compounds and serine esterase inhibitors. In addition to the secretion of preformed mediators, alpha chymotrypsin also induced the metabolism of arachidonic acid, resulting in the release of prostaglandin D2 in a dose-related manner from purified rat peritoneal mast cells. alpha-Chymotrypsin exhibited a marked tissue and species selectivity in its action and tissue mast cells of the rat, guinea pig and human were generally resistant to the enzyme except at cytotoxic concentrations. On the basis of these results, the possible role of endogenous serine esterases in mast cell activation is discussed. PMID- 8720592 TI - Interaction of cytostatics and chemosensitizers with the dexniguldipine binding site on P-glycoprotein. AB - The interaction of cytostatics and chemosensitizers with the dexniguldipine binding site of P-glycoprotein was investigated in photoaffinity labeling experiments. A tritiated azidoderivative of the chemosensitizer dexniguldipine with dihydropyridine structure, [3H]B9209-005, was used to irreversibly label P glycoprotein. The apparent affinity of cytostatics and chemosensitizers to this binding site was estimated from labeling experiments in the presence of increasing concentrations of compounds. From the cytostatics tested, the vinca alkaloids and taxol showed the highest affinity, anthracyclins possessed moderate affinity while methotrexate, ara C and camptothecin, cytostatics not involved in P-glycoprotein-mediated multidrug resistance, were almost inactive. The chemosensitizers GF 120918, cyclosporin A and SDZ PSC-833 inhibited photoincorporation with the highest potency. Steep dose-inhibition curves were obtained with the cyclic peptides and S9788, indicating that these compounds may bind allosterically to a separate binding site. Compounds with dihydropyridine structure with or without chemosensitizing potency were also tested and some structure-activity relationships could be derived from the data. Our data show that inhibition of photoaffinity labeling by [3H]B9209-005 is a valuable and reliable system for measuring the interaction with and potency of chemosensitizing compounds at P-glycoprotein. Furthermore, data obtained in this test system are well suited to investigate structure-activity relationships for chemosensitizers at P-glycoprotein. In addition cytostatics underlying P glycoprotein-mediated multidrug resistance can be identified. PMID- 8720593 TI - Endothelin-1 and angiotensin II act as progression but not competence growth factors in vascular smooth muscle cells. AB - The direct effects of endothelin-1 and angiotensin II on cell cycle progression were investigated in rat aorta smooth muscle cells in primary culture. The phase of the cell cycle was determined by an immunocytochemical analysis of cell cycle specific nuclear antigens. The primary cultured cells were synchronized in the G0 phase (100%) by serum deprivation for 24 h. Endothelin-1 (0.1 microM) or angiotensin II (1 microM) had no effect on the cell cycle of G0 cells, whereas platelet-derived growth factor (PDGF) stimulated the entry of the G0 cells into the G1 phase (100%) without a further progression to the S and M phases. Endothelin-1 or angiotensin II stimulated the progression of the PDGF-pretreated G1 cells to the S and M phases. Fura-2 microfluorometry revealed that, between the G0 and G1 cells, there were no differences in the extent and time course of cytosolic Ca2+ elevations induced by endothelin-1 or angiotensin II, which suggested that endothelin-1 and angiotensin II receptors and their signaling pathways regulating cytosolic Ca2+ remained intact in these cell phases. We thus conclude that endothelin-1 and angiotensin II require the prior G0/G1 transition induced by a competence growth factor such as PDGF to exert their mitogenic effects. These results suggest the important role of endothelin-1 and angiotensin II in atherosclerosis as promoters (progression growth factors), but not as initiators. PMID- 8720594 TI - [3H]Sumatriptan binding sites in human brain: regional-dependent labelling of 5 HT1D and 5-HT1F receptors. AB - The general properties of [3H]sumatriptan binding sites in postmortem human brain tissue sections are described. High concentrations of autoradiographic grains were seen in globus pallidus = substantia nigra > cortex > putamen > hippocampus. While 5-HT (5-hydroxytryptamine) displaced in all regions more than 90% of [3H]sumatriptan binding, the level of binding inhibited by 5-CT (5 carboxamidotryptamine) varied in each region. Although the binding inhibited by 5 CT in some regions such as globus pallidus and substantia nigra was equivalent to that obtained with 5-HT, in cortical areas, such as frontal cortex and hippocampus, a substantial level of binding insensitive to 5-CT was seen. In addition, in membrane binding assays, 10 nM metergoline displaced most [3H]sumatriptan specific binding in striatum and only 16% in frontal cortex. In the human brain sumatriptan binds to at least two 5-HT1 receptors, 5-HT1D and 5 HT1F. PMID- 8720595 TI - Apparent desensitization of a sigma receptor sub-population in neonatal rat cardiac myocytes by pre-treatment with sigma receptor ligands. AB - sigma Receptor ligands induce marked effects on contractility in cardiac myocytes from neonatal and adult rats (Ela et al., 1994, J. Pharmacol. Exp. Ther. 269, 1300-1309; Novakova et al., 1995, Eur. J. Pharmacol. 286, 19-30). Augmentation or attenuation of the contractile amplitude was observed under different experimental conditions. Preincubation of neonatal cardiomyocytes with a sigma receptor ligand ((+)-(3-hydroxyphenyl)-N-(1-propyl)-piperidine ((+)-3PPP), (+) pentazocine, or haloperidol) changed the response to re-application of the ligand after cell wash. The inhibitory effect was abolished, while the stimulatory effect became much more pronounced. We suggest that the effects of sigma receptor ligands are mediated via two receptor subtypes, one stimulatory and the other inhibitory, and only the inhibitory subtype is subject to desensitization. PMID- 8720596 TI - Concerns of an Editor for the Diabetologist. PMID- 8720597 TI - Oral selenate improves glucose homeostasis and partly reverses abnormal expression of liver glycolytic and gluconeogenic enzymes in diabetic rats. AB - Selenium is a trace element that exerts certain insulin-like actions in vitro. In this study, we evaluated its in vivo effects on the glucose homeostasis of rats made diabetic and insulin-deficient by streptozotocin. Na2SeO4 was administered ad libitum in drinking water and/or food for 10 weeks. The elevated plasma glucose levels (approximately 25 mmol/l) and glucosuria (approximately 85 mmol/day) of untreated rats were decreased by 50 and 80%, respectively, by selenate treatment. The beneficial effect of selenate was also evident during oral and intravenous glucose tolerance tests: the integrated glucose responses were decreased by 40-50% as compared to those in untreated rats. These effects were not due to an increase in plasma insulin levels. Compared to non-diabetic rats, pancreatic insulin reserves were reduced by more than 90% in treated and untreated diabetic rats. The hepatic activities and mRNA levels of two key glycolytic enzymes, glucokinase and L-type pyruvate kinase were blunted in diabetic rats. They increased approximately two- to threefold after selenate treatment, to reach 40-75% of the values in non-diabetic rats. In contrast, elevated activity and mRNA levels of the gluconeogenic enzyme, phosphoenolpyruvate carboxykinase, were reduced by 40-65% after selenate administration. Since selenate induced a moderate decrease in body weight due to an anorexigenic effect, we checked that there was no improvement of glucose homeostasis or hepatic glucose metabolism in an additional group of calorie restricted diabetic rats, which was weight-matched with the selenate group. In addition, no obvious toxic side-effects on the kidney or liver were observed in the rats receiving selenate. In conclusion, selenate induces a sustained improvement of glucose homeostasis in streptozotocin-diabetic rats by an insulin like action, which involves partial correction of altered pretranslational regulatory mechanisms in liver metabolism. PMID- 8720598 TI - Suboptimal protein nutrition in early life later influences insulin action in pregnant rats. AB - First-generation rats received either 20% (standard) or 8% (suboptimal) protein nutrition during pregnancy and lactation. Suboptimal protein nutrition led to reduced body weights of the second-generation progeny at day 19 of gestation (10%, p < 0.001) and at weaning (33% reduction, p < 0.001). Control (born of 20% protein-fed dams) and experimental (born of 8% protein-fed dams) offspring received 20% protein diet after weaning and were studied on day 19 of gestation at 9 to 12 weeks after weaning. Basal glucose turnover was lower (29%, p < 0.05) and glucose utilization by fast-twitch muscle, adipose tissue and diaphragm significantly reduced in experimental offspring. Hyperinsulinaemia increased whole-body glucose disposal rate in both control (2.3-fold, p < 0.001) and experimental (3.2-fold, p < 0.001) offspring. Hyperinsulinaemia normalised the suppression of glucose utilization observed in diaphragm, heart and adipose tissue, but not in fast-twitch muscle, where rates remained 30-40% lower in the experimental offspring. Glucose tolerance and insulin secretion after i.v. glucose were unimpaired in the pregnant experimental offspring. A 27% reduction in basal glucose utilization, without impaired growth, was observed for the third generation fetuses of the experimental offspring. The results demonstrate that growth retardation evoked by suboptimal protein nutrition during early life leads to decreased basal glucose turnover and glucose utilization by a range of maternal tissues and the fetus during a subsequent pregnancy. It is not, however, associated with any major permanent impairment of glucose-stimulated insulin secretion or insulin action during pregnancy. PMID- 8720599 TI - Sulphonylurea stimulates glucose uptake in rats through an ATP-sensitive K+ channel dependent mechanism. AB - We studied the effect of gliclazide, a second-generation sulphonylurea, on rat skeletal muscle glucose uptake using perfused hindquarter muscle preparations. Gliclazide at concentrations of 10 to 1000 microgram/ml increased (p < 0.05) the basal glucose uptake. The effect of gliclazide on glucose uptake was immediate and dose-dependent, reaching a plateau at a concentration of 300 micrograms/ml; the half-maximal effect was obtained between 25 and 50 micrograms/ml. The glucose uptake stimulated by gliclazide (300-1000 micrograms/ml) did not differ from that achieved by 10(-9) mol/l insulin, and was lower (p < 0.05) than that obtained with 10(-7) mol/l insulin. The combination of gliclazide (300 micrograms/ml) and 10(-9) mol/l insulin produced an increase in glucose uptake (7.7 +/- 0.6 mumol.g 1.h-1, n = 8, mean +/- SEM) which was higher (p < 0.05) than that achieved with 10(-9) mol/l insulin (5.6 +/- 0.7 mumol.g-1.h-1, n = 11) and not different from that obtained with 10(-7) mol/l insulin (9.8 +/- 1.0 mumol.g-1.h-1, n = 11). Diazoxide (100 mumol/l), an ATP-sensitive K+ channel opener, reversed the stimulatory effect of gliclazide (100 microgram/ml) on muscle glucose uptake from 3.1 +/- 0.4 to 0.5 +/- 0.2 mumol.g-1.h-1, (n = 7, p < 0.001). The addition of diazoxide prior to gliclazide into the perfusion medium blocked the gliclazide induced glucose uptake by the hindquarter muscle preparations. In conclusion, gliclazide alone has an immediate stimulatory effect on glucose uptake by skeletal muscle and together with insulin has an additive effect on muscle glucose uptake. The effect of gliclazide on muscle glucose uptake seems to be due to the inhibition of ATP-sensitive K+ channels. PMID- 8720600 TI - Resistance to tolerance induction in the diabetes-prone biobreeding rat as one manifestation of abnormal responses to superantigens. AB - T cells taken from normal rats treated with an exogenous source of bacterial superantigen in vivo specifically failed to proliferate following re-stimulation with the same superantigen in vitro. Responsiveness was restored following the addition of an exogenous source of interleukin-2 indicating that the T cells had been made functionally tolerant and not deleted. While staphylococcal enterotoxin treatment of normal rats virtually abolished T-cell proliferation to the same enterotoxin in vitro, T cells from similarly treated diabetes-prone Biobreeding (BB-DP) rats were markedly resistant to this in vivo effect. Responses in BB-DP rats were never reduced by more than 50% even when a 4 times more effective dose of enterotoxin was employed. The resistance of BB-DP peripheral T cells to staphylococcal enterotoxin-induced tolerance could not be attributed to differences in T-cell receptor V beta chain family usage of BB-DP vs normal T cells but was associated with qualitative differences in the way in which BB-DP T cells responded to staphylococcal enterotoxins in vitro. While under optimal stimulatory conditions BB-DP T-cell proliferative responses to staphylococcal enterotoxins appeared comparable to those from non-diabetes-prone animals, under superoptimal conditions BB-DP, but not diabetes-resistant, donor T-cell proliferative responses to staphylococcal enterotoxins could be blocked in vitro with antibodies to CD4 antigens. In addition, BB-DP T-cell proliferative responses were more sensitive to suboptimal staphylococcal enterotoxin doses in vitro. We discuss ways in which abnormal BB-DP T-cell responses to superantigens in general and resistance to staphylococcal enterotoxin-mediated tolerance induction in particular may play a role in the generation of a peripheral T-cell repertoire prone to autoimmunity. PMID- 8720601 TI - Function and survival of intrasplenic islet autografts in dogs. AB - Successful transplantation of isolated islets of Langerhans has been reported in large mammals, including man, but metabolic control has not been well established. We studied the glucose and islet hormone response to fasting, i.v. glucose bolus infusion, i.v. arginine bolus infusion during a 35-mmol/l hyperglycaemic clamp, mixed meals, and i.v. insulin-induced hypoglycaemia up to 3 years after intrasplenic islet autotransplantation in six pancreatectomised dogs. The individual postprandial insulinogenic index (ratio of 2-h postprandial insulin to glucose levels) at 1 month post-transplant, predicted (r = 0.99) the time to functional graft failure (6-175 weeks). Metabolic studies at 6 months post-transplant in four dogs demonstrated normal fasting glucose and hormone levels, except for reduced pancreatic polypeptide levels. Intravenous glucose and arginine-stimulated insulin were reduced to 15% of preoperative values. In contrast, postprandial normoinsulinaemia was observed--albeit with moderate hyperglycaemia (approximately 10 mmol/l). Postprandial glucagon and glucose dependent insulinotropic polypeptide (GIP) had increased. Comparison of the post transplant insulin responses to a meal and to intravenous challenges demonstrated maximal stimulation of the graft by the meal. Post-transplant pancreatic polypeptide responses to a meal and i.v. arginine were severely reduced, and no pancreatic polypeptide response to i.v. insulin-induced hypoglycaemia was observed--indicating absence of cholinergic reinnervation. Thus, glucose regulation and both the insulin secretory capacity and life expectancy of islet grafts were best documented by meal testing. Tentatively, a postprandial hyperglycaemia-enhanced incretin effect of glucose-dependent insulinotropic polypeptide and other gut hormones may account for the difference in the insulin response to i.v. glucose and a meal. Aside from the reduced insulin secretory capacity, both a deranged pulsatile delivery of insulin, hyperglucagonaemia, and pancreatic polypeptide deficiency may have been conducive to glucose intolerance. PMID- 8720603 TI - Beta cells are important for islet innervation: evidence from purified rat islet cell grafts. AB - Pancreatic islets receive an extensive and complex innervation that includes sympathetic, noradrenergic nerve fibres also storing neuropeptide Y. Islets transplanted to the kidney capsule become progressively reinnervated, mainly by sympathetic fibres and to a lesser extent by parasympathetic and sensory fibres. The density of nerve fibres in the islet grafts is often higher than in the graft bearing organ, suggesting that the grafted islets contain factors that promote ingrowth of nerve fibres. To find out if beta cells are of any importance for attracting nerve fibres, purified preparations of rat islet beta and non-beta cells were transplanted to the kidneys of nude mice. Some of the mice were rendered diabetic by alloxan injection before transplantation. Immunocytochemical analysis revealed that the beta-cell grafts became richly reinnervated by noradrenergic (tyrosine hydroxylase-containing) nerve fibres, also storing neuropeptide Y. Non-beta islet-cell grafts were virtually devoid of demonstrable nerve fibres. There was no discernible difference in the reinnervation pattern between diabetic and non-diabetic mice. The findings indicate that factors mediating islet neurotrophism are produced by the beta cells. PMID- 8720602 TI - Mechanism of anti-lipolytic action of acipimox in isolated rat adipocytes. AB - Acipimox is commonly used to treat hypertriglyceridaemia in non-insulin-dependent diabetic patients, but its precise mechanism of action has yet to be elucidated. We examined the in vitro effects of acipimox on the lipolytic regulatory cascade in epididymal adipocytes isolated from Wistar rats. Acipimox inhibited the lipolytic rate stimulated by adenosine deaminase (1 U/ml) in a concentration dependent manner, reaching a near-basal value at 10 mumol/l acipimox. Lipolysis activated by sub-maximal levels of isoproterenol in combination with adenosine deaminase (20 mU/ml) was significantly (p < 0.05) decreased by 100 mumol/l acipimox, whereas, in the absence of adenosine deaminase, 100 mumol/l acipimox showed no significant (p > 0.05) inhibition. These findings suggested that the anti-lipolytic mechanism regulated by adenosine may also be regulated by acipimox. Acipimox diminished the intracellular cyclic AMP level produced by 25 nmol/l isoproterenol in the presence of adenosine deaminase (20 mU/ml) in a concentration-dependent manner. At the same level of stimulation, acipimox inhibited the cyclic AMP-dependent protein kinase activity ratio and lipolytic rate over the same concentration range, with significant (p < 0.05) reductions occurring at and above, 0.5 mumol/l and 10 mumol/l acipimox, respectively. Western blotting showed that upon lipolytic stimulation (1 U/ml adenosine deaminase; 100 nmol/l isoproterenol) a threefold increase in the lipolytic rate was accompanied by a significant (p < 0.05) rise in hormone-sensitive lipase associated with the lipid fraction. Acipimox (1 mmol/l) and insulin (1 nmol/l) re distributed hormone-sensitive lipase back to the cytosol, with a corresponding significant (p < 0.05) loss from the fat cake fraction of adipocyte homogenates. In conclusion, the anti-lipolytic action of acipimox is mediated through suppression of intracellular cyclic AMP levels, with the subsequent decrease in cyclic AMP-dependent protein kinase activity, leading to the reduced association of hormone-sensitive lipase with triacylglycerol substrate in the lipid droplet of adipocytes. PMID- 8720604 TI - Elevated serum levels of macrophage-derived cytokines precede and accompany the onset of IDDM. AB - To determine whether cytokines could have a role in the development of insulin dependent diabetes mellitus (IDDM), we measured serum levels of cytokines derived from T helper 1 (interleukin-2 and interferon-gamma), T helper 2 (interleukin-4 and interleukin-10) lymphocytes and macrophages (tumour necrosis factor-alpha, interleukin-1 alpha and interleukin-1 beta) in patients before and after the onset of IDDM. Recently diagnosed IDDM patients had significantly higher levels of interleukin-2, interferon-gamma, tumour necrosis factor-alpha and interleukin 1 alpha than patients with either long-standing IDDM, non-insulin-dependent diabetes (NIDDM), Graves' disease, or control subjects (p < 0.05 for all). Compared with control subjects, patients with long-standing IDDM and those with NIDDM had higher interleukin-2 and tumour necrosis factor-alpha levels (p < 0.01 for all). Interleukin-4 and interleukin-10 were detectable in sera of patients with Graves' disease only, while interleukin-1 beta was not detectable in the serum of any control or test subject. To investigate whether high cytokine levels precede the onset of IDDM, we studied 28 non-diabetic identical co-twins of patients with IDDM, followed-up prospectively for up to 6 years after the diagnosis of the index. Levels of tumour necrosis factor-alpha and interleukin-1 alpha were elevated above the normal range more frequently in the eight twins who developed diabetes than in those 20 who did not (p < 0.005). Analysis of T helper 1 and T helper 2 profiles of the twin groups did not reveal a clear difference between prediabetic twins and twins remaining non-diabetic. These results support the notion that T helper 1 lymphocytes may play a role in the development of IDDM. This is associated with release of macrophage-derived cytokines, which is also a feature of the prediabetic period. The lack of evidence of a dominant T helper 1 profile of cytokine release before diabetes onset suggests that additional events, activating this arm of the cellular immune response, are required in the immediate prediabetic period. PMID- 8720606 TI - Prolonged QT interval as a predictor of mortality in diabetic nephropathy. AB - Patients with diabetic nephropathy face an increased risk of dying due to cardiac causes. The aim of this follow-up trial was to describe the association between the length of the QT interval, as a marker of myocardial electrical stability, and the risk of death in insulin-dependent (IDDM) diabetic patients with overt diabetic nephropathy. A consecutive sample of 85 IDDM patients with overt diabetic nephropathy (i.e. persistent proteinuria > or = 500 mg/24 h) were followed-up until death or for a period of 5-13 years. QT intervals were measured once at baseline in a 12-lead ECG and corrected for heart rate (QTc). During the follow-up period 33 patients (39%) died. In the Cox proportional hazards model independent predictors of death were age (p = 0.0007), the length of the maximum QTc period (p = 0.0049), presence of autonomic neuropathy (p = 0.0068), diabetes duration (p = 0.0163) and RR variation (p = 0.0395). In conclusion, in nephropathic IDDM patients QT prolongation is associated with an increased mortality risk which is independent of the presence of autonomic neuropathy. Further studies are needed to determine whether this risk might be reduced by therapeutic interventions. PMID- 8720605 TI - Does impaired glucose tolerance predict hypertension? A prospective analysis. AB - This study evaluates prospectively the relationship between impaired glucose tolerance (IGT) and blood pressure. From a population of 1376 men and women aged 40-59 years, all those with IGT (n = 54) plus 133 age- weight- and sex-matched normoglycaemic control subjects were selected after excluding treated hypertensive patients. Blood pressure, fasting and postload blood glucose and plasma insulin were measured. At 11.5 years after the first visit 76% of the IGT patients and 80% of the control subjects were re-examined. At baseline blood pressure was significantly higher in IGT patients than in control subjects (systolic 135.5 +/- 2.3 vs 127.9 +/- 1.4 mmHg, p < 0.001; and diastolic 88.0 +/- 1.5 vs 84.7 +/- 0.7 mm Hg, p < 0.05) independent of age, gender, weight, antihypertensive medication and insulinaemia. Accordingly, hypertension was more frequent in subjects with IGT (odds ratio 2.1, 95% confidence, interval (CI) 0.9 4.9). Postload insulin was significantly associated with hypertension--both at univariate and multivariate analysis--in normoglycaemic subjects, but not in those with IGT. At follow-up systolic blood pressure increased in both groups; the increase was smaller in patients with IGT (6.0 +/- 2.4 vs 12.3 +/- 1.6 mm Hg p < 0.05). Likewise, the 11.5 years' cumulative incidence of hypertension was not significantly different in subjects with baseline IGT or normoglycaemia; if anything it was lower in the IGT group (odds ratio 0.36, 95% CI 0.1-1.2). In multivariate analysis incidence of hypertension was associated positively with baseline blood pressure (p < 0.0003) and negatively with IGT status p < 0.03), while no significant association was found with insulin. In conclusion, the findings of this study question IGT as a risk factor for hypertension. Furthermore, these data do not indicate a major role for hyperglycaemia and hyperinsulinaemia per se in the aetiology of hypertension and suggest that IGT and hypertension share one or more pathogenetic factor(s) (i.e., insulin resistance, hyperactivity of the sympathetic nervous system, etc.), which induce deterioration of blood pressure control first, and hyperglycaemia later. PMID- 8720607 TI - Assessment of insulin sensitivity in glucokinase-deficient subjects. AB - The chronic hyperglycaemia of glucokinase-deficient diabetes results from a glucose-sensing defect in pancreatic beta cells and abnormal hepatic glucose phosphorylation. We have evaluated the contribution of insulin resistance to this form of chronic hyperglycaemia. Insulin sensitivity, assessed by the homeostasis model assessment (HOMA) method in 35 kindreds with glucokinase mutations, was found to be significantly decreased in 125 glucokinase-deficient subjects as compared to 141 unaffected first-degree relatives. Logistic regression analysis showed that in glucokinase-deficient subjects a decrease in insulin sensitivity was associated with deterioration of the glucose tolerance status. A euglycaemic hyperinsulinaemic clamp was performed in 14 glucokinase-deficient subjects and 12 unrelated control subjects. In six patients and six control subjects the clamp was coupled to dideutero-glucose infusion to measure glucose turnover. Average glucose infusion rates (GIR) at 1 and 5 mU.kg body weight.min-1 insulin infusion rates were significantly lower in (the glucokinase-deficient) patients than in control subjects. Although heterogeneous results were observed, in 8 out of the 14 patients GIRs throughout the experiment were lower than 1 SD below the mean of the control subjects. Hepatic glucose production at 1 mU.kg body weight-1.min-1 insulin-infusion rate was significantly higher in patients than in control subjects. In conclusion, insulin resistance correlates with the deterioration of glucose tolerance and contributes to the hyperglycaemia of glucokinase-deficient diabetes. Taken together, our results are most consistent with insulin resistance being considered secondary to the chronic hyperglycaemia and/or hypoinsulinaemia of glucokinase-deficiency. Insulin resistance might also result from interactions between the unbalanced glucose metabolism and susceptibility gene(s) to low insulin sensitivity likely to be present in this population. PMID- 8720608 TI - Lack of relationship between muscle sympathetic nerve activity and skeletal muscle vasodilation in response to insulin infusion. AB - Increases in plasma insulin concentration result in vasodilation in skeletal muscle but also in an increase in muscle sympathetic nerve activity (MSNA) which is thought to cause vasoconstriction. The increase in MSNA could therefore be a response to vasodilation (baroreflex), or MSNA could cause vasodilation via putative sympathetic vasodilatory fibres. To examine the relationship between vasodilation, MSNA and insulin action we studied nine non-diabetic Pima Indian men (age 29 +/- 7 years, weight 91 +/- 19 kg, 29 +/- 6% body fat, mean +/- SD) during sequential euglycaemic clamps at low and high insulin doses (80 and 600 mU.m-2.min-1). Leg blood flow was measured by thermodilution, leg glucose uptake by the balance technique, arterial pressure by invasive monitoring and MSNA by microneurography of the peroneal nerve. Whole body glucose uptake (M) ranged from 6.7 to 48.3 during low dose and from 9.4 to 67.7 mumol kg fat free mass-1.min-1 during high dose insulin infusion. At both insulin doses, incremental leg blood flow correlated with M (r = 0.63 and 0.71, respectively). No correlation was found between incremental MSNA and leg blood flow, M or leg glucose uptake. Blood pressure was unchanged throughout the study. MSNA increased after 15-40 min of insulin infusion in all the subjects, whereas leg blood flow started to increase only after 45 min in the most insulin sensitive but not in the most insulin resistant subjects. Thus, insulin stimulates MSNA more rapidly than vasodilation. In conclusion, insulin-mediated MSNA: 1) is neither a response to nor a cause of the vasodilation observed in insulin sensitive men, 2) has no net pressor effect even in the most insulin resistant men in whom insulin-mediated vasodilation was impaired. We conclude that the effect of insulin to stimulate MSNA is dissociated from its acute haemodynamic action. PMID- 8720609 TI - Association between a polymorphism in the angiotensin-converting enzyme gene and microvascular complications in Japanese patients with NIDDM. AB - The relationship between diabetic nephropathy and an insertion (I)/deletion (D) polymorphism in intron 16 of the angiotensin-converting enzyme (ACE) gene is still under debate. The association of ACE gene polymorphism with nephropathy and retinopathy was therefore examined in 362 Japanese patients with non-insulin dependent diabetes mellitus (NIDDM) and 105 healthy control subjects. Distribution of the ACE genotype did not differ between healthy control subjects and diabetic patients without complications. However, the frequency of the D allele was significantly higher in the diabetic subjects with nephropathy than in those without (0.32 in normoalbuminuric patients vs 0.44 in albuminuria patients with albuminuria) (chi 2 = 7.7; p = 0.006). There was no significant association between ACE genotype and retinopathy. These observations thus demonstrate a significant association of the ACE gene polymorphism with nephropathy, but not with retinopathy, in Japanese patients with NIDDM. PMID- 8720610 TI - Effects of an acute decrease in non-esterified fatty acid levels on muscle glucose utilization and forearm indirect calorimetry in lean NIDDM patients. AB - The aim of the study was to evaluate an acute decrease in NEFA levels during an oral glucose tolerance test and its effects on glucose tolerance, muscle glucose uptake and muscle indirect calorimetry in ten lean non-insulin-dependent diabetic subjects. Two 75-g oral glucose tolerance tests were performed in random order. Placebo or 250 mg acipimox (to inhibit lipolysis) were administered orally 2 h before the start of the oral glucose tolerance test. Two hours after acipimox administration (time 0), non-esterified fatty acid, glycerol and 3 hydroxybutyrate levels decreased by 84, 68 and 77% respectively, compared to basal levels. Concomitantly, muscle lipid oxidation and non-oxidative glycolysis also decreased significantly. After placebo administration, non-esterified fatty acids, glycerol and 3-hydroxybutyrate and lipid oxidation increased by 29, 28, 106 and 33%, respectively (NS vs basal levels; p < 0.001 vs acipimox). There was a negative rate of net glucose storage (interpreted as glycogenolysis) during post-absorptive conditions and at time 0 after administration of both drugs. After oral glucose tolerance test, the incremental areas of blood glucose and insulin were significantly decreased by 18 and 19% after acipimox compared to placebo. In addition, the ratio between the incremental area of forearm muscle glucose uptake and the insulin levels was significantly increased by 45% during acipimox compared to placebo administration. Glucose oxidation and non-oxidative glycolysis were significantly higher while lipid oxidation was significantly lower after acipimox than after placebo. In conclusion, our study found that in lean non-insulin-dependent diabetic subjects, an acute decrease in non-esterified fatty acid levels improves glucose tolerance, muscle glucose uptake, glucose oxidation and non-oxidative glycolysis, but is unable to normalize glucose storage. PMID- 8720611 TI - Fasting proinsulin and 2-h post-load glucose levels predict the conversion to NIDDM in subjects with impaired glucose tolerance: the Hoorn Study. AB - The aims of the present study were to observe the natural history of impaired glucose tolerance and to identify predictors for development of non-insulin dependent diabetes mellitus (NIDDM). A survey of glucose tolerance was conducted in subjects aged 50-74 years, randomly selected from the registry of the middle sized town of Hoorn in the Netherlands. Based on the mean values of two oral glucose tolerance tests subjects were classified in categories of glucose tolerance according to the World Health Organization criteria. All subjects with impaired glucose tolerance (n = 224) were invited to participate in the present study, in which 70% (n = 158) were subsequently enrolled. During follow-up subjects underwent a repeated paired oral glucose tolerance test. The mean follow up time was 24 months (range 12-36 months). The cumulative incidence of NIDDM was 28.5% (95% confidence interval 15-42%). Age, sex, and anthropometric and metabolic characteristics at baseline were analysed simultaneously as potential predictors of conversion to NIDDM using multiple logistic regression. The initial 2-h post-load plasma glucose levels and the fasting proinsulin levels were significantly (p < 0.05) related to the incidence of NIDDM. Anthropometric characteristics, the 2-h post-load specific insulin levels and the fasting proinsulin/fasting insulin ratio were not related to the incidence of NIDDM. These results suggest that beta-cell dysfunction rather than insulin resistance plays the most important role in the future development of diabetes in a high risk Caucasian population. PMID- 8720612 TI - Interleukin-12 gene expression is associated with rapid development of diabetes mellitus in non-obese diabetic mice. AB - A single dose of cyclophosphamide (250 mg/kg) is known to synchronize and accelerate development of diabetes in non-obese diabetic mice. We have reported previously that cyclophosphamide treatment of 10-week-old female non-obese diabetic mice induces a shift from T-helper type 2 to T-helper type 1 activity in islet lesions. We now show that this shift in regulatory T-cell function is preceded by the expression of interleukin-12 in the islets as well as in the spleen. In the spleen macrophages were identified as the interleukin-12 expressing cell type. At the same time there was little induction of tumour necrosis factor alpha gene expression by macrophages. Since interleukin-12 is well known to drive T-helper cell type 1 responses we assume that interleukin-12 released by macrophages mediates the accelerating effect of cyclophosphamide on islet inflammation in non-obese diabetic mice. mRNA expression of the p40 chain of interleukin-12 in response to cyclophosphamide was not seen in macrophages of Balb/c mice and thus represents an immune abnormality of non-obese diabetic mice favouring T-helper type 1 reactions. PMID- 8720613 TI - Population-based genetic screening for IDDM susceptibility as a source of HLA genotyped control subjects. PMID- 8720614 TI - Identification of the aminoacid polymorphisms in hexokinase II in Japanese subjects. PMID- 8720615 TI - Insulin analogues and carcinoma of the breast. PMID- 8720616 TI - Antibodies to glutamic acid decarboxylase induced by interferon-alpha therapy for chronic viral hepatitis. PMID- 8720617 TI - Interferon-alpha and IDDM: comment. PMID- 8720618 TI - Desmopressin side effects. PMID- 8720619 TI - Lithium pharmacokinetics. PMID- 8720620 TI - High-dose T4 for rapid-cycling bipolar disorder. PMID- 8720621 TI - Update on bereavement risk. PMID- 8720623 TI - Violent methods associated with high suicide mortality among the young. AB - OBJECTIVE: To present rates and trends in suicide methods, and associations between these, in male and female adolescents in the age groups 15 through 19 years and 20 through 24 years in Finland from 1947 through 1991. METHOD: Suicide rates were calculated on the basis of the mean same-age population, and suicide trends by each method were analyzed by using the Poisson regression model. For the rates and trends, 95% confidence intervals based on the Poisson distribution were calculated. RESULTS: The suicide rate revealed a sharp and significant increase between 1965 and 1975 for both male age groups. Firearms and hanging accounted for the majority of that increase. These figures, however, were not accompanied by similar changes in the availability of firearms during that period. Moreover, an increased total suicide rate as well as that by automobile exhaust fumes in young adults after 1982 coincided with a widely presented Finnish movie which featured this method of committing suicide. In young women, however, intake of solids and liquids remained the most common suicide method. CONCLUSION: Suicide rates increased, especially by violent methods, which contributed to the high suicide mortality rate among the young. These methods, except firearms, however, are often difficult to restrict. This fact underlines the importance of the integration of various preventive measures of cooperation between mental health professionals and other authorities and of the recognition and care of young people at high suicide risk. PMID- 8720622 TI - Treatment of autistic disorder. AB - OBJECTIVE: To present an overview of a variety of treatment approaches in individuals with autistic disorder. METHOD: Selected studies and articles are reviewed. RESULTS: In the past three decades, great progress has been made in the treatment of autistic disorder, particularly in the area of education and parental involvement, with the objective to transfer to home and in other situations learning acquired in school. A role for psychoactive agents, when combined with psychosocial treatments, has been identified. CONCLUSIONS: Although considerable advances have been made in a variety of interventions-educational, psychosocial, and biological-knowledge about the comparative and combined efficacy of the various treatment modalities is lacking. From the parents' perspective, particularly, support and continuity of services require improvement. PMID- 8720625 TI - Negative affect and cognitive biases in suicidal and nonsuicidal hospitalized adolescents. AB - OBJECTIVE: To examine the main and interactive effects of suicidal behavior and gender on negative affect and cognitive biases in hospitalized adolescents (N = 228) aged 13 to 18 years, identified as suicide ideators (n = 68), attempters (n = 90), and nonsuicidal (n = 70). METHOD: Standardized measures of negative affect (depression, anxiety, anger) and cognitive biases (hopelessness, self-concept) were administered upon admission. RESULTS: Suicide ideators reported significantly greater hopelessness and poorer self-concept than did nonsuicidal controls; suicide attempters did not differ from either group. Regression analysis indicated that negative affect and cognitive bias variables accounted for 48% of the variability in suicidal ideation; however, only anxiety and depression were uniquely related to suicidal ideation. Females reported greater depression severity and anxiety than did males; no interaction effects were evident. CONCLUSIONS: Suicide ideators and attempters may have different psychological profiles, with the former reporting greater cognitive biases than nonsuicidal adolescents. The findings regarding suicidal ideation support a mediational model in which negative views of the self and the future contribute to negative affect, and ultimately, suicidal ideation. An integrated assessment of suicidal ideation and affective and cognitive variables is recommended for early risk detection and treatment. PMID- 8720624 TI - State and trait anxiety in adolescent suicide attempters. AB - OBJECTIVE: To examine the relationship between anxiety and suicidal behavior in adolescents. METHOD: Forty-six adolescents who had been hospitalized in an inpatient psychiatric unit after a suicide attempt were compared on measures of anxiety and depression with 72 adolescent psychiatric inpatients who had no history of suicide attempts. RESULTS: The suicide attempters exhibited significantly higher levels of both state and trait anxiety. However, when controlling for depression, the attempters did not differ in their level of state anxiety from the nonattempters, but they still manifested significantly higher levels of trait anxiety than nonattempters. CONCLUSIONS: The results suggest that anxiety, both state and trait, is a risk factor for suicidal behavior in adolescents. Yet, only trait anxiety appears to be relatively independent of depression in its effect on suicidal behavior risk. These findings imply that clinicans should take into account anxiety, both state and trait, for assessment and treatment of adolescents at risk for suicidal behavior. PMID- 8720626 TI - Predictive validity of the suicide probability scale among adolescents in group home treatment. AB - OBJECTIVE: To determine the predictive validity of the Suicide Probability Scale (SPS). METHOD: Prospective design, predicting subsequent suicide attempts, suicide verbalizations, and minor self-destructive behaviors from SPS scores at admission to a group home in a sample of 855 adolescents. RESULTS: SPS scores significantly predicted all three suicide-relevant measures. Only 48% of subsequent attempts could have been predicted from SPS criteria that would have put 21% of newly admitted adolescents in the at-risk group. Although the prediction is far from perfect, it can prevent some suicide attempts when combined with effective suicide precautions. CONCLUSIONS: This is the first known evidence of predictive validity of any measure of suicide risk in adolescents. Its predictive ability may have been reduced by interventions designed to reduce suicide risk. Despite the large number of false-positives and modest predictive validity, a large body of literature indicates that quantitative predictions perform better than clinical judgments in predicting psychological outcomes. However, the SPS measures only a subset of factors predictive of suicide, including suicidal ideation, hopelessness, and social isolation. Risk factors not assessed by the SPS include previous suicide attempts, conduct disorder, substance abuse, and reasons for living. These additional risk factors may be important to improve upon the predictive validity of the SPS. PMID- 8720627 TI - Long-term behavioral sequelae of prematurity. AB - OBJECTIVE: Longitudinal evaluation of the competence and the prevalence of behavior problems in preterm children with serious neonatal complications. METHOD: Prospective follow-up of nonhandicapped preterm children who had been hospitalized in a neonatal intensive care unit (n = 177). The follow-up extended from early school age to early adolescence and was conducted with the help of the Child Behavior Checklist (CBCL) (parent form). RESULTS: The preterm children had lower scores on the Social and School Competence scales than did controls (n = 276). They also were more likely to attend special schools than were children in the general population. With regard to behavior problems, the preterm children had more social problems. The very preterm children and the children who were small for gestational age (SGA) were the ones who contributed to the significant findings. The children who were appropriate for gestational age did not differ from controls. No differential changes in CBCL ratings were found between the preterm and control children. The stability with regard to internalizing problems, attention problems, and social problems was high among the very preterm children and SGA children. CONCLUSION: Very preterm and preterm SGA children are at increased risk of problems in social functioning and functioning at school. These problems persist with age. PMID- 8720628 TI - Resilience and vulnerability among preschool children: family functioning, temperament, and behavior problems. AB - OBJECTIVE: To examine the effects of child temperament and stressful family functioning on child behavior problems among preschool children. METHOD: One hundred forty-five preschool children, aged 2 to 5 years, were evaluated by teachers, mothers, and independent observers. Teachers reported on child temperament; from these ratings, two dimensions of temperament were derived: difficult/easy and approachability. Mothers reported on two dimensions of family functioning: conflict and expressiveness. Both teachers and independent observers rated child behavior problems. RESULTS: Children with more difficult temperaments who were in high-conflict families had the most internalizing and externalizing behavior problems, while children with easy temperaments had fewer such problems, regardless of levels of family conflict. Difficult children whose families were highly expressive engaged in the most observed aggression. CONCLUSIONS: Results suggest that temperament is involved in both protective and vulnerability processes. A difficult temperament operates as a vulnerability factor for internalizing and externalizing behavior problems and observed aggression, while an easy temperament functions as a protective mechanism for these outcomes. PMID- 8720629 TI - Effect of concentration problems on the malleability of children's aggressive and shy behaviors. AB - OBJECTIVE: Previous research has demonstrated the central role of early childhood concentration problems in the development of aggression and other maladaptive behaviors. The present study investigated the moderating effect of concentration problems on the impact of a classroom-based preventive intervention directed at aggressive and shy behaviors in an epidemiologically defined sample of 1,084 urban first-grade children. METHOD: Concentration problems, aggressive behavior, and shy behavior were assessed by a structured teacher interview (the Teacher Observation of Classroom Adaptation-Revised) in the fall and spring of first grade. RESULTS: Children with high ratings on concentration problems in the fall had higher levels of teacher-rated aggressive and shy behavior in the spring than did children without such problems. The intervention reduced aggressive and shy behavior in children regardless of fall concentration level. Boys, but not girls, in the intervention condition with high concentration problems had higher levels of spring aggression than those without such problems, but they also showed the greatest reductions in aggressive behavior from fall to spring. CONCLUSIONS: These results suggest that aggressive behavior is malleable in children with concentration problems, provide further evidence on the etiological significance of concentration problems for the development of maladaptive behavior, and highlight the importance of directly targeting concentration problems to maximize preventive intervention impact. PMID- 8720630 TI - Prevalence rates and correlates of psychiatric disorders among preschool children. AB - OBJECTIVE: To determine the prevalence and correlates of psychiatric disorders among preschool children in a primary care pediatric sample. METHOD: In a two stage design, 3,860 preschool children were screened; 510 received fuller evaluations. RESULTS: For quantitative assessment of disorder (> or = 90th percentile), prevalence of behavior problems was 8.3%. "Probable" occurrence of an Axis I DSM-III-R disorder was 21.4% (9.1%, severe). Logistic regression analyses indicated significant demographic correlates for quantitative outcomes (older age, minority status, male sex, low socio-economic status, father absence, small family size) but not for DSM-III-R diagnoses. Maternal and family characteristics were generally not significant. Child correlates included activity level, timidity, persistence, and IQ. CONCLUSIONS: Overall prevalence of disorder was consistent with rates for older children; correlates varied by approach used for classification. PMID- 8720631 TI - Help-seeking for behavior problems by parents of preschool children: a community study. AB - OBJECTIVE: To identify barriers to seeking help for preschool behavior problems and understand the pattern of service utilization. METHOD: Altogether, 320 preschool children from eight preschool centers were studied using a two-stage design. After the initial screening, a more detailed assessment was carried out in the second stage involving semistructured interviews with parents and children. Parents also completed a service utilization questionnaire during the first stage and General Health Questionnaire, Family Assessment Device, and life events questions during the second stage. RESULTS: Only 19% of those with preschool behavior disorder crossed all the filters in reaching for help. The most common perceived barriers to help-seeking were that problems would get better by themselves or that parents should be strong enough to handle them. The major blocks to help-seeking were at two levels, in parents recognizing the presence of a problem and in overcoming the perceived barriers by the parents. Parents sought help from the informal agencies more often than from the formal. Help was sought significantly less often by those who had parental separation, low income, or multiple adversities, all of which were known to be significantly associated with behavior disorder. CONCLUSION: These findings indicate the need to educate and influence the parents' attitude to help-seeking, target those at risk to develop behavior disorder, and develop better consultation-liaison service with the informal agencies. PMID- 8720632 TI - Effects of Special Olympics International on social competence in persons with mental retardation. AB - OBJECTIVE: To evaluate the social and emotional goals of Special Olympics International, specifically whether Special Olympics facilitates social competence and self-esteem in persons with mental retardation. METHOD: Findings were "triangulated" across three studies on the social competence, adaptation, and self-perceptions of 104 athletes from 1993 Team USA (mean age = 22 years; mean IQ = 59). Study 1 related behavior to athletes' length of time in Special Olympics. Study 2 compared Team USA to an appropriately matched group of non Special Olympians. Study 3 assessed Team USA before and 4 months after their participation in the World Games held in Salzburg, Austria. RESULTS: Relative to age and IQ, length of time in Special Olympics was the most powerful predictor of social competence. Special Olympics athletes had higher social competence scores and more positive self-perceptions than the comparison group. Team USA's behavioral data remained stable over time, suggesting that high pre-Game scores were not simply a function of parental or athlete pre-Game excitement. CONCLUSIONS: Based on the most conservative meaning of triangulation, more support was found linking Special Olympics to social competence than to remaining behavioral domains. PMID- 8720633 TI - Moral valuation: a third domain of conscience functioning. AB - OBJECTIVE: To assess development of moral valuation in normal children and adolescents, that is, how moral rules for living are derived and justified, and to examine the relationship of this progression with previously identified stages of conceptualization of conscience. METHOD: Using three semistructured questions from the Stilwell Conscience Interview, 132 normal volunteers between the ages of 5 and 17 years were assessed. All moral valuation responses were examined within three aspects of social reference: authority-derived, self-derived, and peer derived. Each aspect was scaled for complexity into six anchored levels. RESULTS: The levels of all three aspects correlated positively with conceptualization stages as well as with each other. When the covariate, age, was taken into consideration, peer-derived valuation was significantly correlated with both age and stage. CONCLUSIONS: Moral valuation is a domain of conscience functioning in which moral rules and their justifications are socially referenced in relationship to authority, self, and peers. Anchored levels of these three aspects of moral valuation provide developmental guidelines for mental status examinations in patients between 5 and 17 years of age as well as providing criteria for future comparative studies in various diagnostic categories of psychopathology. PMID- 8720634 TI - Premigratory expectations and postmigratory mental health symptoms in Vietnamese Amerasians. AB - OBJECTIVE: To determine the relationship between premigratory expectations for one's future life in the United States and postmigratory symptoms of anxiety and depression in a group of Vietnamese Amerasians. METHOD: A cohort of 161 Vietnamese Amerasian migrants was assessed prior to departure from Vietnam using the Hopkins Symptom Checklist-25 (HSCL-25), the Vietnamese Depression Scale, and an Expectations Questionnaire. After migration to the United States, subjects were reassessed using the HSCL-25 and the Vietnamese Depression Scale. RESULTS: Premigratory expectations for support from the Vietnamese community in the United States were associated with significantly higher scores on the HSCL-25 Depression scale. Of subjects reassessed in the United States, 20% scored in the clinical range for depression. CONCLUSIONS: Like-ethnic community support is critically important in preventing depressive symptoms among Vietnamese Amerasian migrants. Clinicians working with Amerasians should target those with unrealistically high expectations for preventive intervention and should approach Vietnamese community leaders to mobilize support for recently arrived Amerasians. PMID- 8720635 TI - Resolved: autistic children should have a trial of naltrexone. PMID- 8720636 TI - Reversing a history of unmet needs: approaches to care for persons with co occurring addictive and mental disorders. AB - Individuals with co-occurring addictive and mental disorders are particularly vulnerable to negative outcomes. Historically, they have been treated either in mental health or addiction service settings and sometimes excluded from receiving any services at all. This article presents an overview of the prevalence and clinical correlates of co-occurring disorders, and of the historical development of separate care systems for mental and addictive disorders. The articles that follow address future treatment and service system development. PMID- 8720637 TI - The resurrection and the life. AB - This first-person account of a woman undergoing manic-depression concurrent with alcohol and drug problems highlights the difficulties of attaining help from a system geared toward treating one or the other but not both, and hints at the positive outcomes possible when an integrated approach to co-occurring addictive and mental disorders--one that incorporates the knowledge and perspectives of the client and the family as well as the professional--is available. PMID- 8720638 TI - The epidemiology of co-occurring addictive and mental disorders: implications for prevention and service utilization. AB - General population data from the National Comorbidity Survey are presented on co occurring DSM-III-R addictive and mental disorders. Co-occurrence is highly prevalent in the general population and usually due to the association of a primary mental disorder with a secondary addictive disorder. It is associated with a significantly increased probability of treatment, although the finding that fewer than half of cases with 12-month co-occurrence received any treatment in the year prior to interview suggests the need for greater outreach efforts. PMID- 8720639 TI - Heterogeneity of person and place: assessing co-occurring addictive and mental disorders. AB - Provision of appropriate treatment for clients with co-occurring addictive and mental disorders is hampered by difficulties in diagnosing this diverse population and in identifying and delivering necessary services via agencies that typically focus on only a portion of these clients' problems. This paper considers common pitfalls in assessment across settings and suggests approaches to identifying and meeting the needs of clients with multiple problems. PMID- 8720640 TI - The course, treatment, and outcome of substance disorder in persons with severe mental illness. AB - Individuals with co-occurring substance abuse and severe mental illness are particularly vulnerable to negative outcomes. This paper reviews findings on the longitudinal course of dual disorders in traditional treatment systems, which provide separate mental health and substance-abuse programs; describes the movement toward programs that integrate both types of treatment at the clinical level; reviews evidence related to outcomes in integrated treatment programs; and discusses health-care policy changes that would encourage effective treatments. PMID- 8720641 TI - Co-occurring addictive and mental disorders among adolescents: prevalence research and future directions. AB - Recent epidemiological research documenting the pervasive co-occurrence of addictive and mental disorders has been concerned primarily with adults. This paper proposes the need for similar studies of adolescents, considers the special problems inherent in the assessment of co-occurrence in this age group, reviews evidence suggesting that the prevalence of co-occurring disorders in adolescents parallels that documented for adults, and delineates future research strategies. PMID- 8720642 TI - Women with co-occurring addictive and mental disorders: an emerging profile of vulnerability. AB - The heterogeneity of those with co-occurring addictive and mental disorders has only recently begun to be recognized, and treatment strategies for different segments of this population are still being developed. This article reviews the literature on alcohol and drug problems in women, and on women with severe mental illness who are at high risk for substance abuse--as well as other forms of abuse and deprivation--due to poverty and victimization. As public health and mental health agendas are threatened by budget cuts, it is critical that initial gains in acknowledging and addressing their needs not be lost or abandoned. PMID- 8720643 TI - A vision for the future: toward a service system responsive to those with co occurring addictive and mental disorders. AB - Identified by providers, family members, administrators, and consumers as an issue creating frustration, high costs, and a profoundly negative impact on quality of life, co-occurring addictive and mental disorders cry out for creative and alternative clinical responses. With empirical research and clinical experience supporting the effectiveness of integrated approaches, the time has come to reconsider the systemic division of addictive and mental health services. A change toward integrated systems of care is likely to benefit the mental health and addiction treatment needs of all people, not just those with co-occurring disorders. PMID- 8720644 TI - Preventing violence in a structurally violent society: mission impossible. AB - The futility of violence prevention efforts by government, professional organizations, and social advocacy movements should not be surprising, since the aim of these efforts has been primarily to control, punish, and modify the behavior of individuals involved in violence, rather than to discern and eliminate its root causes in the fabric of societies. This paper argues that efforts to prevent violence will continue to fail, as long as its sources in oppressive and unjust social conditions and values are not understood, confronted, and eradicated. PMID- 8720645 TI - Unrecognized language and speech deficits in preadolescent psychiatric inpatients. AB - Of 55 psychiatrically hospitalized preadolescents with DSM-III-R diagnoses that are not commonly associated with language deficits, 60% were determined to have language or speech deficits, although only 38% had ever received speech or language therapy. The clinical implications of these findings are discussed and guidelines for practice outlined. PMID- 8720646 TI - Parents of children with attention-deficit/hyperactivity disorder: psychological and attentional impairment. AB - Biological parents of ADHD children were compared with parents of nonclinical children on measures of psychological impairment and attentional performance. Parents of children with severe ADHD demonstrated greater impairment in social and psychological functioning. However, no significant differences were found on laboratory measures of sustained attention, memory, cognitive flexibility, encoding, or impulsivity. The possibility that parental adjustment would be more usefully assessed by measures of current adaptive functioning than by means of laboratory tests is discussed. PMID- 8720647 TI - Adolescents with mental retardation: perceptions of sexual abuse. AB - Reactions of nonretarded female undergraduates to sexually coercive situations in which neither, one, or both protagonists had mental retardation were examined. Results indicated that retardation affected perceptions of both responsibility and harm. Implications for the education of mental health professionals about the emotional needs of individuals with retardation are discussed. PMID- 8720648 TI - Acculturation style and psychological functioning in children of immigrants. AB - Thirty-four immigrants, predominantly Asian, and their 48 children were assessed for relationship of acculturation style to children's psychological functioning. Children of parents accepting the majority culture scored higher in social competence. Many of the children reported as having extreme behavioral problems had rejected their ethnic culture. PMID- 8720649 TI - Impact of a natural disaster on preschool children: adjustment 14 months after a hurricane. AB - Fourteen months after a hurricane, young children who had experienced the storm showed significantly higher anxiety and withdrawal and more behavior problems than did children who had not. Behavioral problems decreased steadily over the six months following the storm. Mothers' distress in the hurricane's aftermath was associated with the longevity of their children's emotional and behavioral difficulties. PMID- 8720650 TI - Divorce mediation and resolution of child custody disputes: long-term effects. AB - Separated parents randomly assigned to either mediation or traditional adversarial methods for resolving child custody disputes were surveyed nine years postsettlement. Noncustodial parents assigned to mediation reported more frequent current contact with their children and greater involvement in current decisions about them. Parents in the mediation group also reported more frequent communication about their children during the period since dispute resolution. PMID- 8720651 TI - Cessation and persistence of wife assault: a longitudinal analysis. AB - Patterns of wife assault, rates of cessation, and the effect of cessation on psychological aggression were evaluated in a three-year longitudinal study of 772 couples. More than one-third of the men who had been violent in the year prior to the initial survey ceased or interrupted their violence and showed significant reductions in their psychological aggression during the course of the study. Implications for the conceptualization and further study of wife assault are discussed. PMID- 8720652 TI - War-induced psychic trauma: an 18-year follow-up of Israeli veterans. AB - Eighteen years after their participation in the Yom Kippur War, the psychiatric status of two groups of Israeli veterans--112 combat stress reaction casualties and 189 comparable controls--was assessed. Casualties had higher rates and greater intensity of posttraumatic stress disorder than did controls, both initially and at 18-year follow-up. Similarly, intrusion and avoidance tendencies and psychiatric symptomatology were evidenced more often by combat stress reaction casualties than by controls. Clinical implications of the findings are discussed. PMID- 8720653 TI - Sexual torture as an instrument of war. PMID- 8720654 TI - The spiritual challenge of violent trauma. PMID- 8720655 TI - The effect of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) on cultured dog pancreatic duct epithelial cells. AB - To study the morphologic and genetic events associated with the carcinogenic process in the pancreas, we have isolated and cultured a cell line of dog pancreatic duct epithelial cells and treated these cells with a carcinogen. The pancreatic duct epithelial cells were plated onto Vitrogen-coated Transwell inserts suspended above a feeder layer of human gallbladder myofibroblasts. The epithelial cells grew steadily into polarized monolayers, could be passaged repeatedly, and demonstrated the typical morphologic, immunohistochemical, and flow cytometric profile of normal well-differentiated columnar pancreatic epithelial cells. After being treated with 10(-5) M N-methyl-N'-nitro-N nitrosoguanidine (MNNG) for 48 h, the treated cells grew on plastic surfaces. When grown in organotypic culture, the MNNG-treated cells were cuboidal with a multilayered, pseudostratified architecture. Flow cytometry demonstrated aneuploidy and a high percentage of the cells in S phase after reaching confluency, in sharp contrast to untreated cells. Cytogenetic analysis of the MNNG-treated cells revealed frequent chromosomal trisomy and tetrasomy. The secretion of mucin was also different in the MNNG-treated cells versus the untreated cells. The cultured pancreatic epithelial cells may be useful as an assay system to study the genotoxicity of known and potential carcinogens. PMID- 8720656 TI - Study of pancreatic secretion after subtotal gastrectomy followed by different methods of reconstruction. AB - To study the effect subtotal gastrectomies have on exocrine pancreatic function, 12 dogs were prepared with gastric and duodenal cannulas using a modified technique of Thomas. In our study protocol, we collected pancreatic juice by selective ductal cannulation after having applied two types of stimulation [tryptophan intraduodenally together with secretin intravenously (i.v.) and cholecystokinin i.v. together with secretin i.v.]. Our results did not show a significant increase in pancreatic juice after performing the gastrectomies. In both cases the concentration of bicarbonate decreased significantly in contrast to that in the control group. After Billroth I anastomosis, however, a significant decrease in the first periods of stimulation in relation to the gastrojejunostomy was appreciated. In the case of proteins, a loss of the dose/effect relation in these parameters was seen only after Billroth II anastomosis. PMID- 8720657 TI - Intraabdominal hemorrhage complicating surgical management of necrotizing pancreatitis. AB - Surgical management of necrotizing pancreatitis (NP) may result in significant intraabdominal hemorrhage requiring intervention. To determine the incidence and management of hemorrhage complicating operative management of NP, we analyzed retrospectively all patients undergoing operative treatment of NP between 1985 and 1994. Thirteen of 61 patients (21%) developed intraabdominal hemorrhage requiring intervention. The five patients (38%) who experienced more than one bleeding episode had undergone more prior operative debridements (mean of 5.6 vs. 3.8), had had higher transfusion requirements during the first bleeding episode (mean of 27.4 vs. 11.3 U of packed red blood cells), and had a higher hospital mortality (60 vs. 38%) compared to patients with only one bleeding episode. Coexistence of pancreatic and/or gastrointestinal fistula was more common in patients who developed bleeding (36 vs. 11%). Seventeen bleeding sites (eight venous, seven arterial, two generalized oozing) were identified. Angiography was successful in one of two patients in whom it was employed. Surgical control was effective in the other 12 patients. There was no acute mortality related to hemorrhage, but the hospital mortality was greater than in those without hemorrhage (46 vs. 21%). We conclude that significant hemorrhage complicates the surgical management of NP in approximately 20% of patients; while it can be effectively controlled surgically and does not lead to immediate mortality, it may predict worse prognosis. PMID- 8720658 TI - Cigarette smoking: an independent risk factor in alcoholic pancreatitis. AB - It is not known whether cigarette smoking plays a role as a risk factor in alcoholic pancreatitis. The aim of this study was to compare drinking and smoking habits in three groups of male subjects with an alcohol intake in excess of 40 g/day: (i) 67 patients with acute alcoholic pancreatitis, without other known potential causative agents; (ii) 396 patients with chronic alcoholic pancreatitis; and (iii) 265 control subjects randomly selected from the Verona polling lists and submitted to a complete medical checkup. The variables considered were age at onset of disease, years of drinking and smoking, daily alcohol intake in grams, number of cigarettes smoked daily, and body mass index (BMI). Cases differed from controls in daily grams of alcohol, number of cigarettes smoked and BMI (Mann-Whitney U test, p < 0.00001 for each comparison). Multivariate logistic regression analysis, comparing acute and chronic cases, respectively, versus controls, revealed an increased relative risk of pancreatitis in the two comparisons, associated in both cases with a higher alcohol intake (p < 0.00001) and cigarette smoking (p < 0.00001). No significant interaction between alcohol and smoking was noted, indicating that the two risks are independent. In conclusion, in males a higher number of cigarettes smoked daily seems to be a distinct risk factor in acute and chronic alcoholic pancreatitis. PMID- 8720659 TI - Two forms of hereditary chronic pancreatitis. AB - We report 11 families of hereditary pancreatitis characterized by the presence of calculi in pancreatic ducts. These were classified as (1) calcic lithiasis (one family with five cases), in which the calculi are composed of >95% calcium salts; and (2) protein lithiasis in 10 families, in which the calculi are composed of degraded amorphous residues of lithostathine, the pancreatic secretory protein that inhibits calcium salt crystallization. In both forms, transmission appears to be dominant. The average age at clinical onset of symptoms is 15 years. The clinical progression seems to be less severe than in alcoholic chronic pancreatitis (alcoholic calcic lithiasis). This report shows for the first time that hereditary chronic pancreatitis is a group of at least two diseases having a similar clinical picture and pathological features but different chemical compositions of calculi. This leads us to propose a revised Marseille-Rome classification. PMID- 8720660 TI - Nonsurgical treatment of acute necrotizing pancreatitis. AB - Our objective was to analyze conservative management in a large series of acute necrotizing pancreatitis (ANP) patients in a specialized center for pancreatic disease in the Internal Medicine and Emergency Department of a 1,500-bed hospital. One hundred ninety-nine patients with ANP were considered (mean age, 55.4 +/- 18.2 years; biliary etiology, 60.3%). Identification of necrotizing forms of acute pancreatitis was obtained by ultrasonography, computed tomography scan, and surgery (53 cases). Exclusion of infection of the necrosis was based on clinical findings and percutaneous aspiration with bacteriological sampling. Medical treatment was started in all cases; 37 patients required ICU treatment; 87 patients underwent early endoscopic cholangiopancreatography (ERCP), and endoscopic sphincterotomy was performed in 61 cases; and percutaneous guided aspiration of necrosis and fluid collections was performed in 78 cases and in 9 patients with acute pseudocysts. Fifty-three patients (26.6%) underwent surgery; all patients (n = 30) with infected necrosis were in this group. One hundred forty-six patients were unoperated (73.3%): clinical resolution of ANP and biochemical normalization was achieved in 132 cases (66.3%) at discharge. Ultrasonographic follow-up documented morphological resolution of necrosis and fluid collections in 84 cases (57.5%) at different periods from the onset of ANP (range, 2.2-63.5 weeks); 16 patients dropped out and seven patients are still being followed. Spontaneous regression of acute pseudocysts was observed in 59.2%; percutaneous/endoscopic aspiration was successful in 33.3%. Mortality was 9.5%, significantly lower than the mortality observed in operated patients (28.3%; p = 0.002). Conservative treatment of ANP with sterile necrosis may allow healing in most cases, independent of the extent of necrosis or its association with major complications. PMID- 8720661 TI - Secretin-pancreozymin test (SPT) and endoscopic retrograde cholangiopancreatography (ERCP): both are necessary for diagnosing or excluding chronic pancreatitis. AB - Results of the SPT and the ERCP staged for their severity were compared in 202 patients. The correlation between both investigations was significant (p < 0.001); however, ERCP showed significantly more severe changes (p = 0.04). Furthermore, we found that 129 (64%) patients had parallel SPT and ERCP results, matching in all four gradings of severity. Forty-three (21%) patients had abnormal results for both SPT and ERCP, but the severity gradings did not parallel. Finally, 30 (15%) patients showed totally nonparallel results, a normal SPT and abnormal ERCP, or vice versa. Abnormal ERCP but normal SPT results were found in 23 of these 30 patients (group 1), and normal ERCP but abnormal SPT results in the seven remaining cases (group 2). In the first group, more patients had a history of acute pancreatitis compared to the second group (19 vs. one, p < 0.005). Based on medical history, laboratory and functional test results, and other morphological tests, chronic pancreatitis was diagnosed in two of 23 patients in group 1 and in all seven patients in group 2. Follow-up interviews (86 +/- 54 months) were possible in 20 of the remaining 21 patients in group 1 and showed definite chronic pancreatitis in one and probable chronic pancreatitis in another two of them, whereas in the other 17 patients no symptoms of acute pancreatitis or abdominal pain suggestive of chronic pancreatitis had occurred. In conclusion, both SPT and ERCP should be used to complement each other when chronic pancreatitis is suspected. ERCP seems to over-diagnose the disease since duct changes may only reflect scars after severe acute pancreatitis, or old age, and are not necessarily a sign of chronic pancreatitis. SPT seems to diagnose chronic pancreatitis with more reliability. PMID- 8720662 TI - Serum amino-terminal propeptide of type III procollagen levels in chronic pancreatitis. AB - The severity of pancreatic fibrosis, a characteristic feature of patients with chronic pancreatitis (CP), can be assessed only by direct histologic analysis of pancreatic tissue. Since serum levels of the amino-terminal type III procollagen propeptide (PIIIP) can reflect the degree of fibrogenic activity in several diseases associated with fibrosis, the current study was aimed at investigating whether PIIIP are increased in chronic pancreatitis, the relationship between PIIIP and pancreatic fibrogenic activity, and the influence of pancreatectomy, pancreatic exocrine function, and duration of disease on PIIIP levels. Serum PIIIP was measured in 18 patients with CP (15 without liver disease and three with cholestasis) and in 21 healthy controls. The effect of pancreatectomy on PIIIP was evaluated in seven patients, in whom PIIIP was measured immediately before and 2 months after surgery. Prolylhydroxylase (PHase) activity as an index of pancreatic fibrogenesis was evaluated in pancreatic tissue from 11 patients who had undergone subtotal pancreatectomy and from 11 organ donors. The bentiromide (BT)-PABA test as an index of exocrine pancreatic function was measured in all patients. PIIIP was significantly higher in patients who had or had not undergone pancreatectomy (17.3 +/- 4.0 and 25 +/- 11.4 ng/ml, respectively) than in controls (12.3 +/- 3.1 ng/ml) (p < 0.001). PIIIP decreased significantly after pancreatectomy (before, 32.0 +/- 9.3 ng/ml; after, 18.4 +/- 4.8 ng/ml; p = 0.005). PHase was significantly higher in patients (773 +/- 250 cpm/mg protein) than in controls (405 +/- 121 cpm/mg protein) (p < 0.001). PIIIP was correlated with pancreatic PHase (r = 0.7, p = 0.001) but not with BT-PABA or with the duration of the disease. In conclusion, serum PIIIP levels are increased in patients with CP and reflect the severity of pancreatic fibrogenic activity. No relationship between the serum PIIIP levels and the pancreatic exocrine function and duration of disease was found. PMID- 8720663 TI - Biphasic effect of prostaglandin E1 on the severity of acute pancreatitis induced by a closed duodenal loop in rats. AB - The effect of prostaglandin E1 (PGE1) on the severity of acute pancreatitis induced by a closed duodenal loop in the rat was tested. PGE1 was administered subcutaneously at various doses (3, 6, 12, and 24 microgram/kg) at hourly intervals, from the induction of acute pancreatitis up to the 24th hour. A saline treated group served as the control. The mortality rate was recorded, and pancreatic histology was evaluated by a scoring system. Serum amylase activity and pancreatic amylase, trypsin, protein, and desoxyribonucleic acid (DNA) contents were determined at 24 h. Administration of PGE1 influenced the severity of acute pancreatitis biphasically. Serum amylase was reduced significantly (p < 0.01) at a dose of 12 microgram/kg/h, but less at 24 microgram/kg/h. Pancreatic weights did not differ in the groups. Pancreatic amylase, trypsin, and protein contents showed significant elevations (p < 0.01), yet the mortality rate was reduced using 6 and 12 microgram/kg/h doses of PGE1 compared to controls, but not at higher and lower doses. The DNA content was significantly higher at the 6 microgram/kg/h dose, compared to the control. The extent of necrosis and the severity of hemorrhage were reduced significantly (p < 0.05) at doses of 6 and 12 microgram/kg/h of PGE1, but less at 24 microgram/kg/h. Leukocyte infiltration was not affected. In conclusion, optimal doses of PGE1 can ameliorate, but higher doses increase, the severity of acute pancreatitis in this experimental model. PMID- 8720664 TI - Effects of MCI-727 on pancreatic exocrine secretion and acute pancreatitis in two experimental rat models. AB - The effects of a newly developed compound having antiulcer action, (Z)-2-(4 methylpiperazin-1-yl)-1-[4-(2-phenyl-ethyl)phenyl]-eth anone oxime hydrochloride monohydrate (MCI-727), on pancreatic exocrine secretion were studied in anesthetized rats and evaluated its preventive and therapeutic effects on acute pancreatitis in two experimental rat models. Intraduodenal administration of MCI 727 [25, 50, or 100 mg/kg body weight (wt)] stimulated a dose-dependent increase in pancreatic juice and bicarbonate output without increasing the protein output or plasma cholecystokinin concentration. MCI-727-stimulated pancreatic exocrine secretion was completely abolished by antisecretin serum but not by the cholecystokinin receptor antagonist loxiglumide (50 mg/kg body wt/h) or cholinergic receptor antagonist atropine (100 mu g/kg body wt/h). In rats with acute pancreatitis induced by four subcutaneous injections of 20 mu g/kg body wt cerulein at hourly intervals over 3 h, MCI-727 administered orally at a dose of 100 mg/kg body wt 30 min before the first cerulein injection significantly reduced the increases in serum amylase and lipase activity and pancreatic wet weight and induced improvements in the results of histologic examination. Moreover, when given 30 min before and 90 min after the first cerulein injection, MCI-727 had even more dramatic protective effects on all these parameters. In addition, even when administered immediately after the last cerulein injection, MCI-727 effectively ameliorated all these alterations of acute pancreatitis. However, MCI-727 had no apparent beneficial effects on the biochemical and histologic alterations of acute pancreatitis in the severe form induced by retrograde intraductal injection of 1.0 ml/kg body wt of 4% sodium taurocholate. These findings suggest that oral administration of MCI-727 stimulates pancreatic exocrine secretion by endogenous secretin release and that it has therapeutic as well as preventive effects on mild forms of acute pancreatitis in rats. PMID- 8720665 TI - The involvement and sources of active oxygen in experimentally induced acute pancreatitis. AB - The involvement of active oxygen has been suggested in the development of cerulein-induced acute pancreatitis in rats. Previously, we directly detected pancreatic active oxygen (O2-) production in rats with cerulein-induced pancreatitis by using a supersensitive photon counter and a cypridina luciferin analogue (MCLA) that reacts specifically with O2- by emitting luminescence. In the present study, with the specific aim of determining the source of O2-, we prepared two groups of animals with cerulein-induced pancreatitis: those treated with allopurinol, a xanthine oxidase inhibitor; and those treated with nitrogen mustard, a leukopenia-inducing substance. In each of these two groups, pancreatic O2- production and the severity of pancreatic injuries were comparatively studied. In the leukopenic animal group, decreases in O2- dependent chemiluminescence and improvement in the pancreatic condition coincided. This suggests that neutrophils might be involved in experimentally induced pancreatitis as a source of active oxygen. PMID- 8720666 TI - Calcium channel blockers in experimental acute pancreatitis: effect on tissue prostanoids and oxygen free radicals. AB - The effect of verapamil administration on the changes of prostanoid synthesis, and on free radical production associated with acute pancreatitis, has been evaluated. A necrohemorrhagic model of pancreatitis was induced in Wistar rats by intraductal administration of sodium taurocholate (3.5%). This model is associated with initial increases in prostanoid synthesis and peroxidative damage. Verapamil, administered before pancreatitis induction, prevented initial increases in 6-keto prostaglandin F1alpha (PGF1alpha) and thromboxane B2 (TXB2) but had no effect on PGF2alpha or PGE2 or on lipoperoxidative damage. These results indicate that verapamil administration prevents the increases in pancreatic vasoactive prostanoids (TXB2 and 6-keto PGF1alpha) without affecting the increased levels of PGE2 and PGF2alpha and has no effect on oxygen free radical production in the initial stages of experimental acute pancreatitis. PMID- 8720667 TI - LFA-1 (CD11a/CD18) and ICAM-1 (CD54) antibodies attenuate superoxide anion release from polymorphonuclear leukocytes in rats with experimental acute pancreatitis. AB - The inhibitive effects of anti-CD11a/CD18 (LFA-1) and anti-CD54 (ICAM-1) antibodies on the generation of superoxide anion (O2-) by polymorphonuclear leukocytes (PMNs) was elucidated in rats induced with experimental acute pancreatitis. We investigated the generation of O2- by PMNs in two protocols: in the first, we measured the active oxygen-producing ability of PMNs isolated from blood in normal rats; in the second, we measured it from blood, peritoneal cavity, and bronchial alveolar lavage fluid in rats 3 h after the induction of pancreatitis. In normal rats, although LFA-1 antibody attenuated the generation of O2-, ICAM-1 antibody did not. However, in pancreatitis rats, both LFA-1 and ICAM-1 antibodies reduced the generation of O2- by PMNs isolated from blood and the peritoneal cavity. These results showed not only that both LFA-1 and ICAM-1 antibodies have a protective effect on the generation of O2-, but also that LFA-1 has a direct inhibitive effect on the generation of O2- by PMNs in this model. Furthermore, histological studies showed there to be less neutrophil accumulation in the lungs of LFA-1- and ICAM-1-treated animals compared to control animals. PMID- 8720668 TI - All-trans-retinoic acid inhibits growth of human pancreatic cancer cell lines. AB - Retinoids are a class of molecules structurally related to vitamin A that have potent antiproliferative and differentiating effects on a variety of normal and neoplastic tissues. All-trans-retinoic acid (ATRA) has become a first-line chemotherapeutic agent in the treatment of certain leukemias; however, the effect of ATRA on pancreatic tumors is unknown. The purpose of this study was to determine the effect of ATRA on the growth characteristics of both exocrine and endocrine human pancreatic cancer cell lines. The in vitro growth of four cell lines was examined after treatment with a wide dose range of ATRA. The growth of all tumor cell lines was inhibited by ATRA in a dose-dependent fashion beginning at 0.1 microgram M. The in vivo growth of functioning human pancreatic carcinoid (BON) xenografts in Balb/c athymic mice was determined by treatment with several doses of ATRA over 1 month. The growth of BON tumors was inhibited in a dose dependent fashion. These results suggest that ATRA exerts direct antiproliferative effects on both exocrine and endocrine human pancreatic cancers and may be useful in the chemotherapy of these tumors. PMID- 8720669 TI - Combined analysis of expression of c-erbB-2, Ki-67 antigen, and tenascin provides a better prognostic indicator of carcinoma of the papilla of Vater. AB - Expression of the molecular biological factors (MBFs) c-erbB-2, Ki-67 antigen, and tenascin (TN) was assessed immunohistochemically in specimens from patients who had undergone surgery for carcinoma of the papilla of Vater. The MBFs were then analyzed by histological factors (v, d, panc, n, Stage), which have been demonstrated to be outcome predictors, and by patient outcome. None of the MBFs showed any significant correlation with the histological factors. There were significant differences (p < 0.05) in the expression of c-erbB-2, Ki-67 antigen, and TN between patients who survived >5 years and those who survived <5 years. The patients with greater expression of c-erbB-2, Ki-67 antigen, and TN had a poor prognosis, whereas those with less expression had a good prognosis. They were therefore considered independent predictors of outcome for carcinoma of the papilla of Vater. Combined analysis of both histological factors and MBFs was also performed, with the result that the combined analysis of MBFs yielded a better prediction of outcome in carcinoma of the papilla of Vater than analysis of either one histological factor or MBF. PMID- 8720670 TI - An unusual case of chronic pancreatitis of possible immune origin. PMID- 8720671 TI - Pancreatitis and an abnormal sweat test at presentation in a child with non Hodgkin's lymphoma. PMID- 8720672 TI - Acute pancreatitis after ingestion of acetic acid. PMID- 8720673 TI - Induction of diabetes in animal models for pancreatic islet research. PMID- 8720674 TI - Glucagon-like peptide-1(7-36)amide increases cyclic AMP accumulation in normal islets. PMID- 8720675 TI - Alterations in anterior segment dimensions in eyes with retinitis pigmentosa. AB - BACKGROUND: In visually deprived primate eyes there is a correlation between alterations in growth parameters and changes in retinal neurotransmitters. Progressive retinal degeneration also leads to alterations in retinal neurotransmitters. In this study we wanted to investigate whether alterations in the dimensions of the anterior eye segment are also present in human donor eyes of patients with hereditary retinal degeneration. METHODS: Sagittal sections through anterior segments of 47 eyes from donors with different types of retinitis pigmentosa (RP) and of 59 eyes from normal donors were investigated morphometrically using a PC-based morphometric system. RESULTS: In the RP eyes there was a significant shortening of the distance between the peripheral end of Descemet's membrane and the posterior end of Schlemm's canal and of the length of the pars plana of the ciliary body. These changes varied markedly among the different types of RP. The most pronounced shortenings were found in eyes of men with X-linked recessive RP, which had the most severe retinal changes. RP eyes showed, in contrast to normal eyes, significant age-related elongation of the ciliary body and the chamber angle. A significant increase of the axial diameter with age was also found in RP eyes. CONCLUSION: Our study reveals for the first time that there are changes in the anterior segment dimensions in human eyes with hereditary retinal degeneration. Measurements of anterior eye segment parameters might provide additional indices for the severity of RP and for the diagnosis of RP subtypes. PMID- 8720676 TI - Indirect immunofluorescence study of the cornea and the conjunctiva in a xenogeneic system: analysis of humoral reactivity in patients with Wegener's granulomatosis. AB - BACKGROUND: Ocular involvement in patients with Wegener's granulomatosis (WG) can cause progressive loss of vision. The pathophysiological events leading to the WG specific ocular inflammation are unknown. METHODS: A method involving indirect immunofluorescence on bovine and porcine corneoconjunctival frozen sections was used to detect antibodies to corneal and conjunctival epithelium in serum samples from patients suffering from WG with ophthalmic involvement (orbital granuloma or ocular vasculitis, both active and inactive phase). Normal persons and WG patients without ophthalmic involvement served as control groups. Anti-human IgG, IgA and IgM were tested. RESULTS: Five different immunofluorescence patterns could be distinguished in each species. There were differences in the reactivity with bovine and porcine tissue. Every pattern was caused mainly by human IgG. The antibodies were directed to the cell nuclei or to extranuclear structures. All patterns also occurred frequently in normal sera. No pattern was found that occurred specifically in the serum of patients with WG and ocular inflammation. CONCLUSIONS: The results indicate that this immunofluorescence system does not detect a characteristic antibody profile that could be attributed to the serum of WG patients. Hence the approach lacks clinical usefulness as a diagnostic procedure. PMID- 8720677 TI - The development of central areolar choroidal dystrophy. AB - BACKGROUND: Central areolar choroidal dystrophy (CACD) is a hereditary macular disorder of which the development is poorly understood. METHODS: One hundred and eight members of seven families with CACD underwent ophthalmological examination. If macular alterations were found or suspected, the patients underwent fluorescein angiography, electroretinography (ERG), electrooculography (EOG) and tests of colour vision and visual field. CACD was divided into four stages: I, slight parafoveal changes of the pigment epithelium (RPE); II, RPE mottling encircling the fovea; III, additional atrophy of the choriocapillaris without central involvement; IV, as stage III with central involvement. RESULTS: In 60 eyes of 30 patients, 8 with stage I, 12 with stage II, 18 with stage III and 22 with stage IV CACD were found. The photopic ERG was subnormal in about half of the cases with stage II-IV. Colour vision tests revealed diminished red sensitivity and pseudoprotanomaly in stages I and II and combined red-green and blue-yellow defect in stages III and IV. Parafoveal reduced sensitivity (stages I and II) and parafoveal and foveal reduced sensitivity (stages III and IV) were found in the visual field tests. CONCLUSION: We describe and expand the stages of development of CACD. Early recognition of patients may have a great influence on their subsequent life. PMID- 8720678 TI - Retinal and disc neovascularization in Behcet's disease and efficacy of laser photocoagulation. AB - BACKGROUND: The vaso-occlusive episodes resulting from Behcet's disease can cause capillary dropout and vascular remodeling. Retinal and disc neovascularizations, which occur as a result of occlusive vasculitis, can cause recurrent vitreal hemorrhages and neovascular glaucoma leading to severe visual impairment. METHODS: 1080 eyes of 540 patients with Behcet's disease were examined between 1973 and 1993. Of the 912 eyes with posterior segment involvement, laser photocoagulation could be performed in 13 of 25 eyes with disc neovascularization (NVD), 12 of 22 eyes with retinal neovascularization (NVE), and 4 of 6 eyes with NVD and NVE. Laser was directed at areas of NVE and retinal capillary nonperfusion. In cases of NVD, panretinal photocoagulation was performed. RESULTS: The rate of regression of NVD was significantly greater in laser-treated eyes than in the untreated group. The results were similar in cases of NVD with NVE. In eyes with NVE which underwent laser photocoagulation, the NVE regressed. None of the treated eyes developed neovascular glaucoma during the follow-up period. Vitreous hemorrhage occurred in two laser-treated eyes. CONCLUSION: Laser photocoagulation is successful in preventing complications of retinal and disc neovascularizations. Thus, in cases of occlusive vasculitis associated with Behcet's disease, laser photocoagulation should be considered for prevention of complications such as vitreous hemorrhage and neovascular glaucoma. PMID- 8720679 TI - Immunoscintigraphy for ocular melanoma: a reliable diagnostic technique? AB - BACKGROUND: Immunoscintigraphy (IS) has recently been used as a diagnostic tool for ocular melanoma. We wanted to reevaluate published data in our own patients and to correlate immunoscintigraphic results with histologic findings and immunohistochemical characteristics of the tumour tissue. METHODS: During a 4 year period, IS was performed on 35 patients (average age 64 years) with suspected ocular melanoma by i.v. injection of 225.28S, a monoclonal antibody against high-molecular-weight melanoma-associated antigen. Histology was available in 22 cases. Tumour tissue was evaluated for cell type, vascularization, necrosis, pigmentation, and lymphocytic infiltration, and immunohistochemistry was performed with 225.28S and antibodies against HMB-45, S 100 and vimentin. One hundred and two patients with metastasizing cutaneous melanoma served as controls. In these patients the identical immunoscintigraphic technique was applied. RESULTS: IS yielded a positive result in about 50% of our patients with ocular melanoma, while in patients with cutaneous melanoma sensitivity was 89%. In five patients who turned out not to have melanoma, two false-positive results were obtained (one subretinal hemorrhage and one Wegener's granulomatosis). No correlation was found between any of the histological features or the immunoreactivity pattern and the immunoscintigraphic outcome. However, antigenic differences between ocular and cutaneous melanoma were evident. CONCLUSION: We conclude that IS, using the antibody applied in this study, is of only limited value in patients with ocular melanoma. Our results suggest that antigenic differences, rather than histological characteristics or technical problems, are responsible for the low sensitivity in ocular melanoma compared to cutaneous melanoma. PMID- 8720680 TI - Proteinases in subretinal fluid. AB - BACKGROUND: Degradation of the extracellular matrix by secreted proteases is connected to cell migration and proliferation in invasive growth and in scar tissue formation. In retinal detachment, retinal pigment epithelium (RPE) cells loosened from their monolayer are often seen in the subretinal fluid (SRF) and the vitreous, where they may participate in the scar tissue formation of proliferative vitreoretinopathy. To evaluate the role of SRF constituents on the release of RPE cells, we analyzed SRF in patients with retinal detachment for the presence of enzymes able to degrade extracellular matrix. METHODS: SRF was collected altogether from 16 patients undergoing retinal reattachment surgery and analyzed for activities against some of the key enzymes in extracellular proteolysis, namely collagenases, gelatinases, elastase and cathepsin G. RESULTS: Seventy-two-kilodalton gelatinase was found in all SRF samples studied, whereas the neutrophil-type 92-kDa gelatinase could not be detected. Low collagenase, elastase and cathepsin G activities could also be detected in some samples. CONCLUSIONS: The predominant type of matrix metalloproteinase present in SRF is the 72-kDa MMP-2. The proteolytic activity in SRF may be connected to the release of RPE cells into SRF and to degradation of components of the vitreous exposed to SRF. PMID- 8720681 TI - Functional neuroanatomy of visual object naming: a PET study. AB - BACKGROUND: The ability to name objects depends partly on visual perception. We used positron emission tomography (PET) to measure activity-related changes in regional cerebral blood flow (r-CBF) in order to identify regions of the brain activated during visual object naming. METHODS: Four right-handed volunteers were recruited. Following an intravenous injection of 15O-labeled water, r-CBF was measured during visual object naming, counting numbers, and resting. PET and MRI images were coregistered, the size of the brain was proportionally adjusted in each axis to Talairach's and Tournoux's atlas, and the comparison of stimulated versus resting blood flow images revealed activated brain regions. RESULTS: In the subtraction of resting from naming, activation was observed in the bilateral primary visual cortex, bilateral fusiform gyrus, left lingual gyrus, bilateral inferotemporal cortex, bilateral inferior frontal gyrus, bilateral precentral gyrus, anterior cingulate gyrus, left parietal operculum, and left putamen. In the subtraction of counting from naming, most of the those areas were activated, but no significant activity was observed in the left lingual gyrus, left parietal operculum, or bilateral precentral gyrus (motor cortex). The areas activated with the paradigm included those dedicated to visual perception (primary and associate visual cortex), visual recognition (inferior temporal cortex), and phonological output (Broca's area). CONCLUSION: Our results indicated that the major neural network from occipital lobe to frontal cortex, which is mainly involved in the ventral visual pathway, demonstrated activation in these tasks. Result of this study will serve as base line data for analyzing the findings in patients with impaired visual perception. PMID- 8720683 TI - Vasoactive intestinal peptide- and nitric oxide synthase-containing nerve fibers in the rat ophthalmic artery have different origins. AB - BACKGROUND: Neuropeptides are important in the regulation of vascular functions. The purpose of the present study was to investigate the distribution of perivascular nerve fibers containing neuropeptides and nitric oxide synthase (NOS) in the rat ophthalmic artery. METHODS: Neuropeptide- and NOS-containing nerve fibers in the rat ophthalmic artery were identified using immunocytochemistry following unilateral parasympathectomy (electrocoagulation of the sphenopalatine ganglion) and sympathectomy (superior cervical ganglionectomy). The contralateral artery was used as control. RESULTS: Nerve fibers containing vasoactive intestinal peptide (VIP), neuropeptide Y (NPY) and NOS were abundant in the wall of the ophthalmic artery. Nerve fibers containing calcitonin gene-related peptide and substance P were scarcer. Following destruction of the sphenopalatine ganglion there was no discernible change in the density of VIP- and NPY-immunoreactive fibers, while the majority of the NOS containing fibers disappeared. Following sympathectomy the NPY-containing nerve fibers were almost completely eliminated, whereas VIP- and NOS-containing fibers were unaffected. CONCLUSION: The present study demonstrates that the sphenopalatine ganglion is the main contributor of NOS-containing nerve fibers to the rat ophthalmic artery. In contrast, this ganglion apparently is of little importance as a source of VIP-immunoreactive nerve fibers to the artery, demonstrating different origins of the majority of the VIP-and NOS-containing fibers to this artery. The NPY-containing nerve fibers in the rat ophthalmic artery emanate mainly from the superior cervical ganglion. PMID- 8720684 TI - Pyogenic granuloma of the cornea: report of a case and review of the literature. AB - BACKGROUND: Pyogenic granulomas are ulcerated vascular proliferations, often accompanied by inflammatory infiltrates, which rarely affect the eye. Only few reports on pyogenic granulomas of the cornea have been presented. METHODS: The enucleated globe containing the corneal tumor was examined histopathologically, and special immunohistochemical stains were performed. RESULTS: The lesion had a yellowish appearance with marked vascularization. The inflammatory infiltrate was mainly composed of mononuclear cells, with no multinucleated giant cells, and was located anterior to Bowman's layer, leaving the corneal stroma unaffected. Immunohistochemical studies outlined the high vascular content, even in dense cellular areas, and confirmed the inflammatory nature of the mononuclear infiltrates. CONCLUSIONS: Pyogenic granuloma of the cornea represents a clinical and morphological entity developing in a traumatized eye with corneal vascularization. Although the origin of these lesions remains obscure, there have been no signs of malignancy. Therefore, enucleation of an eye with a pyogenic granuloma should be avoided. PMID- 8720685 TI - Midline granuloma presenting as orbital cellulitis. AB - BACKGROUND: Lethal midline granuloma usually presents with rhinorrhoea and redness of the skin above the nose. Early ocular symptoms are very rare. We here describe a patient who presented with acute orbital cellulitis. PATIENT: A 73 year-old woman had a 24-h history of severe pain around her left eye. We saw the typical clinical picture of orbital cellulitis. A CT scan revealed a diffuse infiltration of the left upper and lower lid, the anterior orbit and the ethmoidal sinuses. RESULT: On surgical exploration we found a granular, partly necrotic tumour. Histological examination revealed an angiocentric nasal T-cell lymphoma (midline granuloma). CONCLUSION: Midline granuloma should be included in the differential diagnosis of acute orbital cellulitis. PMID- 8720682 TI - The influence of penetrating keratoplasty and cyclosporin A therapy on MHC class II (Ia)-positive cells in the rat iris and choroid. AB - BACKGROUND: The presence of Ia-positive cells (MHC class II equivalent) has been previously reported in the iris and choroid of various species. They have been reported to have both round and dendritic morphologies; the latter may represent classic dendritic cells, potent antigen-presenting cells (APCs). It is possible that the dendritic-like cells play a important role in (auto)immune processes of uveal and other ocular tissues. Using the flat or whole mount technique, the distribution of Ia-positive cells in the rat iris and choroid was investigated following penetrating keratoplasty (PKP) and following treatment with cyclosporin A (CsA). METHODS: Lewis (LW) rats received corneal buttons from Lewis-Brown Norway (LW-BN) donors and were randomly assigned to the following groups: (i) operated, untreated (n = 24); (ii) operated, CsA-treated (10 mg/kg i.m.; n = 22). Controls were groups (iii) normal LW rats (n = 13); (iv) unoperated, CsA-treated (16 days' treatment; n = 8); (v) anterior perforation of the anterior chamber (n = 3); (vi) eight corneal sutures only (n = 4); (vii) syngeneic operated (LW to LW; n = 4). Animals of groups (i) and (ii) were killed on the 5th, 9th and 13th postoperative days and on appearance of the corneal rejection (group i, day 13; group ii, day 16). Both eyes were enucleated, immediately fixed, and iris-choroid flat mounts were examined for Ia-positive cells using APAAP immunohistochemistry. RESULTS: In the normal Lewis rat iris, scattered Ia-positive cells of both nondendritic and dendritic morphology were observed. CsA treatment in the unoperated rat did not result in a significant decrease in the percentage of dendritic cells in the iris or choroid. Anterior chamber perforation, the placement of sutures in the cornea and syngeneic PKP resulted in a moderate increase in iris Ia-positive cells. Allogeneic transplantation resulted in a large increase in both types of Ia-positive cells, particularly on day 13 with corneal rejection. In group ii, an initial decrease in Ia-positive cells until day 13 was observed; upon rejection (day 16), the histological picture was similar to that of untreated animals. Alterations in the operated choroid were also apparent following CsA treatment. CONCLUSION: Corneal transplantation in the Lewis rat results in an increase in Ia-positive cells in the iris; CsA therapy can delay but not prevent this reaction. Changes in choroidal Ia-positive cells following PKP were not apparent, their numbers being affected only by CsA treatment following grafting. PMID- 8720686 TI - Properties of intracellular calcium stores and their role in receptor-mediated catecholamine secretion in rat adrenal chromaffin cells. AB - Simulation of rat adrenal chromaffin cells either with receptor agonists such as muscarine, bradykinin and histamine or with caffeine in Ca(2+)-free Krebs solution induced a brief increase in the intracellular free Ca2+ concentration, [Ca2+]i, which terminated within 90 s. Both the agonist- and caffeine-induced [Ca2+]i responses were abolished in cells which had been treated with either 500 nM thapsigargin (TG) or 20 microM ryanodine, suggesting that inositol trisphosphate and caffeine cause Ca2+ release either from the same Ca2+ store or from distinct stores which exchange Ca2+ rapidly. In normal Krebs solution, these agonists evoked catecholamine (CA) secretion which showed an initial transient followed by a sustained component. Neither component of the secretion was significantly affected by TG or ryanodine. In the medium containing 16 microM Ca2+ or no Ca2+, CA secretion evoked by 30-second stimulation with 100 microM muscarine was 59 or 7%, respectively, of that evoked in the normal medium containing 2 mM Ca2+. In TG-treated cells, the CA secretion at 16 microM Ca2+ was reduced to 23% and that in the Ca(2+)-free medium was completely abolished. These results suggest that the receptor-mediated Ca2+ entry solely determines the rate of CA secretion in rat chromaffin cells when stimulated by receptor agonists in the normal medium, whereas intracellular Ca2+ release and Ca2+ entry may cooperatively support the secretion when cells are stimulated in media containing low concentrations of Ca2+. PMID- 8720687 TI - Angiotensin II changes inositol-1,4,5-trisphosphate content in the pituitary and hypothalamus but not in cerebral cortex of the rat brain. AB - The present study was undertaken to investigate the effect of angiotensin II (AngII) on the concentration of inositol-1,4,5-trisphosphate (IP3) and the modulation of AngII action by estradiol in the brain. Our studies were conducted in male rat anterior pituiary, hypothalamus and cerebral cortex with and without previous estradiol treatment. In the cerebral cortex, there were no changes in IP3 content after exposure to AngII. In the anterior pituitary, AngII increased the IP3 concentration at all doses studied. However, in the hypothalamus, AngII decreased the IP3 concentration and its effect was also time dependent. Prior treatment with estradiol intensified the effects of AngII action on IP3 concentrations, in both the pituitary (increase) and hypothalamus (decrease). These results indicate that in the male rat anterior pituitary, AngII increases phosphatidiloinositol hydrolysis, leading to the increased production of the intracellular messenger, IP3. The decrease of IP3 concentration observed in the hypothalamus suggests a different site of AngII action in this region. We also suggest that estrogens can modulate AngII action in the brain. PMID- 8720688 TI - Regional and diurnal studies of melatonin and melatonin binding sites in the duck gastro-intestinal tract. AB - Since melatonin and putative melatonin receptors can be detected in a variety of peripheral tissues, direct endocrine and paracrine actions of melatonin on the physiological functions of different organ systems in response to internal and external stimuli probably exist. As an extension of our earlier work on the pharmacological characterization of 2-[125I]iodomelatonin binding sites in the duck jejunum, the regional and diurnal variations of melatonin and putative melatonin receptors of different segments of the duck gastro-intestinal tract were studied in an attempt to understand the role of melatonin in the physiology of the digestive system. Although no significant effects of diurnal variation and pinealectomy on the regional distribution of melatonin were observed, significant regional variations of melatonin levels were detected with decreasing levels as follows: colon > oesophagus, caecum > duodenum, jejunum > ileum. The densities of melatonin binding sites showed a significant variation between different intestinal regions at either mid-light or mid-dark, with the following descending order: ileum, jejunum > duodenum, colon > caecum > oesophagus. Analysis of the distribution of melatonin binding sites in the wall of the intestine revealed maximal binding in the mucosa and minimal binding in the muscular layers of the jejunum. Similar results were obtained for other intestinal regions as revealed by autoradiography. No significant changes in the affinities of melatonin binding sites were detected between different regions and tissue layers of the alimentary canal. Moreover, the densities and affinities of melatonin binding sites among different regions of the gut exhibited no significant diurnal variations. The demonstration of regional variations in melatonin levels and the density of melatonin binding sites along the gastro-intestinal tract, with a concentration of the putative melatonin receptors in the mucosal layer, suggests a possible direct action of melatonin in the regulation of fluid/electrolyte transport and nutrient absorption in the gut. PMID- 8720690 TI - Hormonal regulation of plasminogen activator in rat and mouse seminiferous epithelium. AB - To elucidate the possible role of plasminogen activator (PA) in spermatogenesis and spermiation in mammals, we studied the hormonal regulation of PA secretion in cultured rat and mouse seminiferous tubules during defined stages of spermatogenesis. Results indicated that: (1) under basal conditions, segments of rat seminiferous tubules released primarily urokinase-type PA (uPA) at all stages of the cycle. The highest level of PA secretion occurred at stages VIIab, VIIcd and VIII. FSH, 8-bromo cyclic AMP and forskolin (FK) stimulated PA secretion, predominantly tissue-type PA (tPA). (2) In contrast, mouse seminiferous tubules secreted only tPA under basal conditions. In the presence of 50 microM MIX, seminiferous tubules at stages VII and VIII secreted higher levels of both types of PA than at the other stages. Both tPA and uPA secretion was enhanced by addition of FSH and FK to the organ culture media. (3) Segments of both rat and mouse seminiferous tubules at stages IX-XII in which the sperm residual bodies are absorbed into the Sertoli cells were also very sensitive to the addition of FSH to the organ culture. These results suggest that tPA in rat and mouse testes may play an essential role in the process of spermatogenesis and spermiation as well as in sperm residual body absorption. PMID- 8720689 TI - Effects of melatonin on hypothalamic gamma-aminobutyric acid, aspartic acid, glutamic acid, beta-endorphin and serotonin levels in male mice. AB - Effects of melatonin on hypothalamic neurotransmitters in male mice were studied. Exogenous melatonin administered intraperitoneally significantly increased (p < 0.05) hypothalamic concentrations of aspartic acid and gamma-aminobutyric acid by over 29 and 50% respectively. Conversely, hypothalamic beta-endorphin concentration was significantly decreased (p < 0.05) 30 min after melatonin administration with doses between 5- and 100 micrograms/kg. Similarly, melatonin, at a concentration of 100 micrograms/kg, decreased (p < 0.05) the serotonin level in mouse hypothalamus by 46%. Melatonin, however, did not affect the concentration of hypothalamic glutamic acid over a dose range of 0.5-300 micrograms melatonin/kg. Our findings suggested that actions of pineal melatonin in animals such as inhibition on serum corticosterone levels might be mediated by the potentiation of activities of hypothalamic neurons containing gamma aminobutyric acid and aspartic acid or by the inhibition of the beta-endorphin and serotonin hypothalamic neurons. The neurons containing glutamic acid in the hypothalamus were, however, not influenced by melatonin. Our results are in line with the suggestion that melatonin actions on adrenal corticosterone release or other endocrine secretions may be mediated by way of its actions on hypothalamic neurotransmitter activities. PMID- 8720691 TI - Three different gating models for inactivation of the Ito in ventricular myocytes of rats and mice. AB - Kinetic models to describe the time course of the Ca(2+)-independent outward potassium current (Ito) in cardiac ventricular cells were constructed. The Ito traces were recorded from isolated myocytes of rats and mice using the whole-cell configuration of the patch-clamp technique. An iterative method was developed to eliminate the capacitive transient overlapping the early part of the Ito. The isolated Ito curves were then fitted by Hodgkin-Huxley type functions and different inactivation gating mechanisms were identified in various groups of the animals. In some Wistar rats a single inactivation route was found. Another group of Wistar rats and diabetic BB/Mol rats showed two independent inactivation pathways both of which resulted in a completely closed channel. In lean specimen of C-57 mice and in streptozotocin-treated Wistar rats the two inactivation gates gave closed channels only when both parallel inactivating transitions have been completed. In this case a possible interaction between the gating particles has been revealed. PMID- 8720692 TI - Distribution of proteins in aqueous two-phase systems formed by dextran and polyethylene glycol. Influence of protein hydrophobicity. AB - Aqueous two-phase systems formed by mixing polyethylene glycol (PEG) and dextran (DXT) were studied. The composition of DXT and PEG phase was established by the size exclusion HPLC method. The concentrations of DXT and PEG in both phases as a function of total concentration of both polymers was fitted by nonlinear regression. This procedure yielded an empirical numerical model (with two independent variables: bulk concentration of DXT and PEG) characterizing the composition of either phase at any applied concentration of the polymers. The distributions of bovine serum albumin (BSA) and immunoglobulin G (IgG) were studied in this system in relation to the composition of both phases. Both proteins were predominantly distributed in the dextran phase. The protein affinity to the respective phase was found to increase with: i) the increasing phase dextran concentration; and ii) the decreasing phase PEG concentration. Chemical modification of primary amino groups of BSA by 2,4,6-trinitrobenzene sulfonic acid (TNBS) raised the affinity of the conjugate to the less polar PEG phase. PMID- 8720693 TI - Mechanisms of vasoconstriction induced in frog vascular smooth muscle by MD1, a new biotechnological agent. AB - The mechanism of action of MD1, a new biotechnological radiotherapeutic agent of plant origin, on smooth muscle contraction was investigated. Contraction induced by MD1 consists of an initial dose-dependent transient phasic response (PR) (with ED50 = 0.11 +/- 0.02%, n = 4, the time of rise 15 s, onset of decay about 30 s) followed by a tonic contraction (TC) (at 10% MD1 only) with a maximum in 20 min (0.78 +/- 0.07 x the maximum contraction induced by 110 mmol/l KCl, n = 8, (TC/PR) = 0.94 +/- 0.11, n = 5). If the vessel was washed out during this maximum, the maximum tension was maintained for up to 2 h ("long-lasting" tension) and was abolished after perfusion with Ca(2+)-free EGTA solution (E) or H7 (0.05 mmol/l) solution. With solution E being applied 30 min before 10% MD1 induced contraction, TC was reduced by 92 +/- 3% (n = 4) in contrast to PR being reduced by 48 +/- 5% (n = 5). During TC, calcium appeared to penetrate into cells through receptor operated channels, since tension neither depended on verapamil (0.05 mmol/l) nor cobalt (up to 10 mmol/l). In solution H7 (with 10 min pretreatment), PR and TC were almost completely inhibited. It is proposed that MD1 activates the C-kinase branch of the calcium messenger system. PMID- 8720694 TI - Analysis of bursting in Stein's model with realistic synapses. AB - By introducing realistic synaptic potentials to the Stein's stochastic model of neuron we obtain a new model able to produce bursting activity. The mechanism of burst initiation is described and basic characteristics, such as period of bursting, period of quiescence and frequency of spikes are calculated from model parameters. PMID- 8720695 TI - Enzyme kinetics and the activation energy of Mg-ATPase in cardiac sarcolemma: ADP as an alternative substrate. AB - Increasing concentrations of Mg within a range between 0.1-5.0 mmol/l step-by step activated the Mg-dependent ATPase and ADPase in rat heart sarcolemma. Both Mg-dependent activities were influenced by NaN3 in a similar way. Also, activation of both enzymes by their substrates, ADP and ATP, were affected by NaN3 in a similar mode. It appears that both enzyme activities are secured by the same system which is capable of ADP hydrolysis during ATP insufficiency. In the absence of naN3 the enzyme revealed higher affinity to ATP than to ADP. The activation energy was lower for ATP hydrolysis. The above findings indicate that at non limiting concentrations of Mg2+ the enzyme is favoring ATP. PMID- 8720696 TI - The 45Ca2+ uptake by Trichoderma viride mycelium. Correlation with growth and conidiation. AB - The properties of Ca2+ uptake by Trichoderma viride were studied using radionuclide 45Ca2+ in conjunction with the study of effects of agents influencing the Ca2+ homeostasis on the 45Ca2+ uptake, vegetative growth and conidiation. Mycelium of T. viride was found to take up 45Ca2+ in time- and temperature-dependent manner. The 45Ca2+ uptake could be distinguished from the 45Ca2+ binding by the insensitivity to washing with EGTA (ethylene glycol-bis(2 amino ethyl ether)-N,N,N',N'-tetraacetic acid)-containing solution. The 45Ca2+ uptake was only slightly suppressed by the treatment used to de-energize cells. Agents known to influence Ca2+ homeostasis in animal and plant cells were also active in perturbing the Ca2+ homeostasis in T. viride. In this respect, the agents tested had dual (stimulatory or inhibitory) effects on the 45Ca2+ uptake. No clear correlation among the perturbation of the 45Ca2+ uptake and the inhibition of growth and conidiation was found for the group of compounds tested. Sr2+ and Mg2+ inhibited 45Ca2+ uptake but did not inhibit growth and conidiation. Co2+, Cd2+ inhibited both 45Ca2+ uptake and growth. Other agents tested (Cu2+, Ni2+, La3+, dihydropyridines), which inhibited growth of T. viride, induced massive 45Ca2+ uptake by its mycelium. Ba2+ and Mn2+ showed a biphasic effect on 45Ca2+ uptake-inhibition at lower, and stimulation at higher concentrations, but they had only a slight inhibitory effect on the growth or conidiation at higher concentrations. The 45Ca2+ uptake was influenced by addition of monovalent cations to a small extent only. Na+ (up to 75 mmol.l-1), less than K+, slightly suppressed the 45Ca2+ uptake leaving both growth and conidiation unaffected. Upon depriving the fungus of Ca2+ by chelation of extracellular Ca2+ (not Mg2+ or divalent trace metals) by EGTA, which interfered with Ca2+ homeostasis, vegetative growth rate, and starvation-induced conidiation were restricted. These results suggest that the sustained Ca2+ influx occurs across the T. viride plasma membrane which may be a target site for the antifungal action of heavy metal ions, and its perturbation may lead to disturbances in physiological processes including growth and conidiation. The properties of the Ca2+ influx in T. viride observed substantially differ from those observed in animal cells. PMID- 8720697 TI - Cytoplasmic calcium fluctuations in calcium overloaded Xenopus laevis oocytes. AB - Cytoplasmic calcium fluctuations were studied in calcium overloaded Xenopus oocytes. Calcium sensitive chloride currents were recorded using the two electrode voltage clamp technique. Fluctuations of chloride currents measured under the voltage clamp were elicited by injection of calcium into the cytoplasm. Contrary to infrequent injections of small calcium amounts which evoke smooth transient responses, the fluctuating chloride currents are due to overloading of intracellular calcium stores which then release calcium repeatedly. Chloride current fluctuations in calcium overloaded oocytes can be reversibly suppressed by caffeine. This effect is concentration dependent, and the amplitude decrease of fluctuations is already apparent at 2 mmol/l caffeine. Power spectra density of fluctuations have been analyzed; they exhibited this pronounced effect of caffeine. Other effective inhibitors were tetracaine and heparin. The results of the present work suggest that at least a part of the endoplasmic reticulum in Xenopus oocytes is a calcium releasing calcium store which can be activated by calcium at the resting inositol trisphosphate concentration. PMID- 8720698 TI - Effects of haloperidol and chlorpromazine on smooth muscle contractility, platelet aggregation and neuronal calcium current. AB - Effects of chlorpromazine, haloperidol (neuroleptics and calmodulin antagonists), and verapamil on rat platelet aggregation induced by thrombin, on calcium current in snail neurones and on both tonic tension of high potassium contracture and phasic contraction of isolated guinea-pig ureter preparations were studied. Moreover, droperidol, sulpiride and prazosine effects were studied for models of phasic contractility and platelet aggregation. Sulpiride and prazosine were ineffective, verapamil was ineffective on platelet aggregation, while droperidol was the most potent inhibitor of platelet aggregation. These results, the similarity revealed in the blockage of neuronal calcium current by neuroleptics and verapamil, and the potent inhibitory action of haloperidol and chlorpromazine on contractility and aggregation suggest that both phenothiazine and butyrophenone neuroleptics possess some properties of calcium antagonists and may also have intracellular sites of action other than calmodulin. PMID- 8720699 TI - A note on the cellular effects of nystatin in single myoballs. AB - Volume changes in single L6 myoblasts (myoballs) exposed to nystatin solutions were followed on single cell level by means of quantitative video image analysis. The myoblasts swelled in nystatin solutions. The volume change was dependent on the nystatin concentration, the threshold concentration being 12.5 mumol/l of nystatin freshly dissolved in Krebs solution. The threshold effect was triphasic: a slight initial volume decrease (shrinkage) for about 2 min followed by a volume increase and, after about 10 min by a significant volume decrease. At twice as high nystatin concentration (25 mumol/l) the final shrinkage phase was lacking. At 50 mumol/l concentration the volume increased continually after a delay of about 1-2 min. and reached a plateau of about 350% of the original volume. At 100 mumol/l concentration of nystatin the myoblasts increased their volume in about five min to more than 500% of the original value. The effects of nystatin diminished upon prolonged storage of nystatin Krebs solution. Nystatin solutions (50 mumol/l) prepared 3 hours before use were stil active to about 80%. Volume changes in 100 mumol/l nystatin solutions were, however, substantially diminished (to about 20%) 5 hours after the preparation of the nystatin solution. By replacing external Na+ by TEA+ in the presence of external Cl- a regulatory volume decrease was observed to subnormal values; the myoblast volume shrank to about half of the control value. The volume changes were reversible after reintroduction of Krebs solution. The regulatory volume decrease to subnormal values was also observed after replacing external Cl- by glutamate anion in the presence of external Na. The volume changes were, however, not reversible after reintroduction of Krebs solution. The swelling of myoblasts in 50 mumol/l nystatin Krebs solution continued after a definite enlargement of the whole myoblast was reached with the formation of several blebs, which eventually coalesced to form a continuous layer around the myoballs. The enlarged vesicles in nystatin solutions were able to start and fulfill the mitotic cycle. Cell volume measurements represent a handy means for checking the activity of nystatin solutions for the perforated patch experiments. PMID- 8720700 TI - [Toxic encephalopathy; occupational disease as a consequence of organic solvents]. PMID- 8720701 TI - [Classification and grading of glial tumors]. PMID- 8720702 TI - [Source and clinical significance of allergy for thiomersal, an organic mercury compound]. PMID- 8720703 TI - [Pseudoxanthoma elasticum; etiology, diagnosis and therapy]. PMID- 8720704 TI - [Favorable results of minimally invasive surgery in the treatment of primary hyperparathyroidism]. AB - OBJECTIVE: To study the appropriateness of minimally invasive surgical treatment of primary hyperparathyroidism. DESIGN: Prospective. SETTING: University Hospital Utrecht, the Netherlands. METHOD: In patients with primary hyperparathyroidism, parathyroid adenomas were located preoperatively by means of Doppler assisted ultrasonography and spiral computer tomography. If the results were positive, minimally invasive surgery was performed. RESULTS: Minimally invasive surgery was carried out in 13 out of 15 successive patients with good results (the serum calcium and parathyroid hormone levels returned to normal). Two patients were subjected to conventional neck exploration, also with good results (preoperatively several adenomas were suspected in one, while no adenoma was seen in the other). CONCLUSION: It can be calculated that minimally invasive surgery will probably suffice in 60-70% of the patients with primary hyperparathyroidism, so that conventional neck exploration can be avoided. PMID- 8720705 TI - [Stroke in the elderly: prevalence and frequency of hospitalization; the ERGO study (Erasmus Rotterdam Health and the Elderly). The ERGO research group]. AB - OBJECTIVE: To evaluate the prevalence of self-reported and medically confirmed stroke, and the degree to which the event leads to hospitalization. DESIGN: Cross sectional study. SETTING: The suburb of Ommoord in Rotterdam. METHODS: The Rotterdam Study is a single centre population-based prospective follow-up study of 7983 subjects, aged 55 years and over, living in the suburb of Ommoord of Rotterdam. Information of stroke was based on the question 'Did you ever suffer from a stroke, diagnosed by a physician'? If yes, additional medical information was obtained from the general practitioner. The present analysis is based on the first 4982 participants of the Rotterdam Study (1878 men and 3104 women) of whom data on both self-reported and medically confirmed stroke were available. RESULTS: Prevalence of self-reported stroke was 5.8% for men and 4.0% for women. Of self-reported strokes 55% could medically be confirmed, yielding a prevalence of 3.5% for men and 1.9% for women. The prevalence of first and only stroke was 4.5% and 3.0% for men and 3.3% and 1.7% for women, respectively and increased with age. The percentage of hospitalization was 55% for self-reported first and only stroke and 61% for medically confirmed first and only stroke. CONCLUSION: Among elderly subjects, self-reported stroke was relatively common and in 55% the stroke was confirmed by medical records. About 55% of the patients with a first stroke were hospitalized. PMID- 8720706 TI - [Henoch-Schoenlein purpura-nephritis followed by IgA-nephritis in 3 children]. AB - In three patients, two boys of 7 and 10 and a girl of 9 years old, a kidney disease was diagnosed which evolved from Schonlein-Henoch nephritis to IgA nephropathy or Berger's disease. These two kidney diseases show striking similarities in pathological and immunological respects. Several patients have been described in the literature who suffered from both clinical pictures consecutively and of families in which different members showed either one or the other disease. A remarkable finding was the persistence of an elevated IgA serum level in all three children after the Schonlein-Henoch nephritis had subsided. These cases show the importance of long-term follow up of patients who had Schonlein-Henoch nephritis, in order to diagnose late renal complications at an early stage. They also give more evidence in favour of the assumption that Schonlein-Henoch nephritis and IgA nephropathy have a common aetiology. PMID- 8720707 TI - [Granulomatous hepatitis attributed to the combination pyrimethamine chloroquine]. AB - After her holiday in South Africa, a 50-year-old woman was admitted because of fever and pain in the upper abdomen. The laboratory tests showed moderately increased serum liver enzyme activities. The liver biopsy showed a granulomatous hepatitis. Further investigations revealed no evidence for sarcoidosis, tuberculosis or infectious hepatitis, nor for other granulomatous diseases or infectious diseases relevant to South Africa. Upon discontinuation of the malaria prophylaxis with Daraclor (pyrimethamine and chloroquine (sulphate)) the symptoms disappeared and the liver function tests returned to normal. It was concluded that Daraclor was the probable cause of granulomatous hepatitis in this patient. This adverse effect was not published before. PMID- 8720708 TI - [Active euthanasia in newborn infants with spina bifida?]. PMID- 8720709 TI - [Quality care in medical education; important role for students and great significance of final terms]. PMID- 8720710 TI - [Pharmacotherapy: clinical practice]. PMID- 8720711 TI - [Systematic approach to a difficult diagnostic problem]. PMID- 8720712 TI - Baseline alterations in blue-on-yellow normal perimetric sensitivity. AB - BACKGROUND: the extent of any learning and fatigue effects in blue-on-yellow (B Y) perimetry is unknown. The within- and between-eye changes in B-Y normal sensitivity at a single visit and between visits was investigated as a function of previous experience in white-on-white (W-W) perimetry and of subject age. METHODS: Sixty-one normal subjects 22 young (range 20-34 years) and 39 elderly (range 51-80 years)-- assigned to one of four groups based upon perimetric experience and age underwent B-Y perimetry on four separate occasions using Program 30-2 of a modified Humphrey Field Analyser 640. Perimetry was performed for both eyes on three consecutive days and again one week later. Global, hemifield and annular mean sensitivities and global short-term fluctuation were calculated for each eye at each visit. RESULTS: Mean sensitivity was higher in the first eye examined and increased over the 3 days. The increase in mean sensitivity was similar for each eye and was independent of perimetric experience and age. Short-term fluctuation decreased over the 3 days, regardless of eye and experience, but was more pronounced for the younger age group. The extent of all improvements varied considerably between subjects. CONCLUSIONS: Improvements in B Y perimetric performance occurred as the familiarity with the test increased, regardless of previous W-W experience. Failure to account for such improvement could reduce the efficiency of B-Y perimetry. PMID- 8720713 TI - Prognosis of orbital lymphoid hyperplasia. AB - BACKGROUND: Orbital lymphoid hyperplasia can be associated with systemic non Hodgkin lymphoma (NHL), even when polyclonal proliferation is found in the orbit. Although irradiation is recommended, some orbital lymphoid hyperplasias are treated by steroids (when inflammation is clinically presumed) or left untreated. METHODS: The incidence of concurrent NHL and the incidence of future NHL after oral prednisone, radiotherapy, or no treatment were retrospectively evaluated in 33 cases of lymphoid hyperplasia (22 benign lymphoid hyperplasias, BLH, and 11 atypical lymphoid hyperplasias, ALH), after follow-up of 2-13 years. RESULTS: NHL occurred in 12 of 33 cases (36.4%). In seven patients it was concurrent; in five patients it occurred 2-6 years later. In the actuarial curve, at 5 years 55% of patients were free of lymphoma, at 10 years, 46%. NHL was more commonly observed when the lacrimal gland was involved (57% vs 21%; P = 0.03). Of the 13 patients treated with oral steroids, 46% had complete response, 39% partial response, and 15% future NHL. Of the seven irradiated patients, five (71%) had complete response, two (29%) partial response, and none future NHL. Of the eight untreated patients, five (63%) had partial response and three (37%) future NHL. Irradiated lacrimal gland BLHs only achieved partial response, one having radiation-induced orbital inflammation. CONCLUSION: Because of a high risk of NHL, in all orbital lymphoid tumors systemic staging and follow-up are mandatory. The advised management is irradiation, except for Sjogren syndrome, an initially inflammatory lacrimal gland BLH, where a course of steroid is suggested before considering radiotherapy. PMID- 8720714 TI - Partial characterization of a putative new growth factor present in pathological human vitreous. AB - BACKGROUND: Several growth factors have been implicated in the development of proliferative eye diseases, and some of those are present in human vitreous (HV). The effects of HV on cellular responses which modulate proliferative cell processes were studied. This study describes the partial characterization of a vitreous factor activity which does not correspond to any of the previously reported growth factors in pathological HV. METHODS: Vitreous humour was obtained from medical vitrectomies, from patients with PDR and PVR. The biological activity of the vitreous factor was determined by its ability to increase cytosolic calcium concentration ([Ca2+]i), increase production of inositol phosphates, and induce cell proliferation in the cell line EGFR T17. In some experiments other cell lines, such as NIH 3T3, 3T3-L1, FRTL5, A431, PC12, Y79, and GH3, were also employed. Measurement of [Ca2+]i in cell suspensions was performed using the fluorescent Ca2+ indicator fura-2. The activity of the factor present in HV was compared with other growth factors by means of: (a) [Ca2+]i mobilization pattern, (b) sequential homologous and heterologous desensitization of receptors, (c) effects of phorbol esters on their action, and (d) inactivation after treatment with different proteolytic enzymes. RESULTS: The HV-induced cell proliferation and increases in [Ca2+]i concentration were characterized by a peculiar time pattern. The different approaches used ruled out its identity with PDGF, bFGF, EGF, TGF-beta, IGFs, TNF-alpha, NGF, and other compounds such as ATP, angiotensin I, and bradykinin. Vitreous factor actions are mediated by specific receptors apparently regulated by PKC. This factor is able to induce [Ca2+]i mobilization in most of the cell lines studied, indicating that its effects are not tissue specific. CONCLUSIONS: These results suggest the presence of a growth factor activity in pathological HV which may be due to the presence of an undescribed growth factor in the eye. PMID- 8720715 TI - Cross-matches on donor cadaver retinal pigment epithelial cells in corneal risk patients. AB - BACKGROUND: Allografts can be rejected either through the antibody-mediated or cellular pathways. The objective of this study was to look at the extent of antibody formation in patients awaiting re-keratoplasty using cross-matches on cadaver retinal pigment epithelial (RPE) cells. METHODS: Cadaver RPE cells were derived by trypsin digestion from donor eyes (n = 1200). After 3 days of cell cultivation, the cells were adherent and began to lose their pigment. By day 7 most cells were clear and grew as a polygonal monolayer. MHC class I expression by RPE cells was studied by the W6/32 (anti-HLA-A, B, C) monoclonal antibody (MoAb) and that of class II (HLA-DR) by the 136 MoAb. Normal RPE cells express few class I and no detectable class II antigens. For the induction of MHC expression, cells were subsequently stimulated with 250 U/ml of recombinant gamma interferon for 5 days. Cells were used for tissue typing and also for cross matches with recipient serum. Cross-matches were subsequently performed and measured by flow cytometry. RESULTS: Both class I and class II antigens were strongly enhanced, as could be shown by immunohistochemical staining. Some 20% of those patients awaiting rekeratoplasty (n = 60) were positive for anti-HLA antibodies. In one case anti-DR3 antibodies were detected in a recipient who had had several rejection episodes after keratoplasty. CONCLUSIONS: RPE cells are not only useful for cadaver post-mortem HLA typing but also for donor-specific cross matches. The degree of antibody formation after keratoplasty in rejecting patients was, however, low. This may imply that anti-HLA antibodies are not the major cause of corneal graft loss after keratoplasty. PMID- 8720716 TI - Pseudoexfoliation syndrome with poorly dilating pupil: a light and electron microscopic study of the sphincter area. AB - BACKGROUND: The study was carried out to examine histological changes in the iris sphincter muscle and surrounding tissue in eyes with pseudoexfoliation syndrome (PXS). METHODS: Seventeen patients with PXS, 14 of them having capsular glaucoma, were enrolled into the study. Iris biopsies were obtained during extracapsular cataract extractions (ECCE) requiring sphincterotomies. Thirteen biopsy specimens underwent histological examination by light microscopy, and four were examined with the electron microscope. Three iris biopsies from cadaver eyes and one obtained during ECCE from a patient with a miotic pupil (no PXS) served as control specimens on light microscopy. Electron microscopic controls included three iris specimens from cadavers. RESULTS: In the light microscopic examination, blood vessel walls were stained with Congo Red in seven of the 13 PXS specimens. In three of these seven specimens the stromal tissue was extensively fibrotized, and in two specimens fibrosis was moderate. Distinct stromal fibrosis was also observed in one and moderate fibrosis in two PXS specimens negative for amyloid. The control specimens were not positive for Congo Red, but stromal tissue was moderately fibrotized in one specimen. Light microscopy did not reveal differences in muscle tissue between PXS and control specimens. Electron microscopically, however, the muscle tissue was fibrotic or disorganized in three PXS specimens and in one control specimen. Fibrils, similar to the fibrillar component of pseudoexfoliation material described in previous studies, were found in intimate association with the capillary basal lamina in all of the PXS specimens showing muscular fibrosis. CONCLUSIONS: These findings support the theory that PXS is associated with amyloid, and in some PXS eyes miosis is connected with degenerative changes both in the stromal tissue and in the muscular layer of the iris. PMID- 8720717 TI - Susceptibility of human retinal pigment epithelial cells to different viruses. AB - BACKGROUND: Different viruses have been reported to be involved in retinal diseases in animal systems. In humans, herpes simplex virus and cytomegalovirus have been found to cause retinal disease. Most of the studied viruses are neurotropic. In this study, the in vitro susceptibility of human retinal pigment epithelial cells (RPEC) to representative members of different groups of human pathogenic viruses was investigated. METHODS: Early cultures of RPE C - after two or three passages - were infected with the following viruses: herpes simplex virus (HSV) type 1, human herpesvirus 6 (HHV-6), Epstein-Barr virus (EBV), cytomegalovirus (CMV), adenovirus types 1 and 7, measles virus, parainfluenza virus and coxsackie virus B3. RESULTS: Cultures of RPE C could be infected with neurotropic viruses like HSV or measles virus as well as with typical respiratory viruses like parainfluenza or adenoviruses. Coxsackievirus, an enterovirus, replicated as well as human CMV, whereas EBV and HHV-6, two lymphotropic viruses, failed to infect RPE. CONCLUSION: These findings suggest that a variety of viruses, including those causing rather common illnesses, might be capable of inducing retinal lesions under certain circumstances due to haematogenous spread during the course of viraemia. PMID- 8720718 TI - Vitamin E succinate inhibits proliferation and migration of retinal pigment epithelial cells in vitro: therapeutic implication for proliferative vitreoretinopathy. AB - BACKGROUND: Retinal pigment epithelial (RPE) cells play an important role in proliferative vitreoretinopathy (PVR). Vitamin E succinate is an ester form of a potent biological antioxidant, vitamin E, and has unique effects on various cells. We examined the effect of vitamin E succinate on proliferation and migration of cultured bovine RPE cells, since these are critical steps in the development of PVR. METHODS: Bovine RPE cells were cultured in minimal essential medium (MEM) containing 10% fetal calf serum (MEM-10). Cells were incubated with MEM-10 containing 25 microM vitamin E, vitamin E succinate, butylated hydroxytoluene (BHT) or d-mannitol. Cell proliferation was assessed by counting cell numbers on days 2, 4 and 6. 3H-Thymidine uptake was also examined in RPE cells incubated with various forms of vitamin E-- vitamin E, vitamin E succinate, Trolox, gamma-tocopherol, vitamin E acetate, vitamin E phosphate, vitamin E nicotinate--or antioxidants-- BHT or d-mannitol (25 microM each). RPE cell migration was studied as follows: A small area (5 x 15 mm) of confluent cultured RPE cells was denuded using a straight razor blade and incubation was continued for 20 h with MEM-10 containing vitamin E, vitamin E succinate, gamma-tocopherol or BHT. The number of cells that migrated into the denuded area from the wound edge in each microscopic field (x20) was counted and expressed as a percentage of control (MEM-10 alone). RESULTS: The antioxidants, vitamin E and BHT, stimulated RPE cell proliferation and 3H-thymidine incorporation compared with the control, while vitamin E succinate significantly inhibited both proliferation and 3H thymidine uptake (IC50, 23 microM). Other forms of vitamin E or d-mannitol had no effect. Neither vitamin E nor BHT had a significant effect on RPE cell migration (108.2% and 112.6% of control, respectively), but vitamin E succinate inhibited migration (58.3%). Cell viability, assessed by the trypan blue dye exclusion test, was not impaired by a 3-day incubation with 50 microM of vitamin E succinate. CONCLUSIONS: An ester form of a physiological antioxidant, vitamin E succinate, inhibits RPE cell proliferation and migration without causing cellular toxicity. These findings suggest its therapeutic potential for the pharmacological treatment of PVR. PMID- 8720719 TI - Production of neocollagen by cells invading hydrogel sponges implanted in the rabbit cornea. AB - BACKGROUND: Poly(2-hydroxyethyl methacrylate) sponges are artificial tissue equivalent matrices with potential value as materials for the peripheral zone of artificial corneas. A keratoprosthetic device was developed incorporating a poly(HEMA) spongy skirt which allowed cellular invasion. The present in vivo study investigated the biosynthetic activity of stromal fibroblasts growing within a poly(HEMA) sponge implanted into the rabbit cornea. METHODS: A porous poly(HEMA) hydrogel was synthesized by polymerization in a large excess of water. Specimens with a pore size larger than 10 microns were impregnated with collagen type I and then implanted into the limbal region of cornea in four rabbits. The animals were followed clinically for 28 days, when they were anaesthetized and new sponge specimens were implanted in their second eye. After 2 h, both eyes were enucleated. The 28-day and 2-h explants were subjected to autoradiographic analysis following labelling with tritiated proline and to an immunostaining technique using antibodies to collagen types I-VI. RESULTS: The autoradiographic analysis showed that the fibroblasts within the 28-day explants continued to be synthetically active and deposited proteins. Using the immunostaining technique, the deposition was most clearly demonstrated by the localization of collagen type III in the tissue invading the sponge. Both techniques failed to indicate any cellular activity in the short-time implants. CONCLUSIONS: The presence of collagen type III is consistent with a normal healing response of the stromal fibroblasts and indicates that poly(-HEMA) sponges are able to function as tissue equivalent matrices. PMID- 8720720 TI - Topography of the "vitreous structures" (tractus; membranelles) with respect to the layers of embryonal blood vessels. AB - BACKGROUND: It is believed that the vascular or primary vitreous, with the exception of the later Cloquet's canal, gradually disappears and is substituted by the avascular or secondary vitreous. It is known that it is possible topographically to objectify mats of fibril concentrations (membranelles or tractus) of stronger light reflection inside the adult vitreous. These concentrations open up in the shape of a funnel from the papilla or Cloquet's canal towards the front of the vitreous. METHODS: This was a light microscopy investigation on human eyes between the embryonal stage of 3.2 cm and the fetal stage of 12.5 cm and eyes 8 months and 2 years old with persistent vessels of the vitreous. RESULTS: The investigation showed that at the embryonal stage, the vitreous body is threaded from behind by branches (mats) of the hyaloid artery and from in front by vessels (mats) that go over the rim of the optic cup, i.e., the later vitreous base. Vitreous structures, in the form of horsetail-shaped fibril concentrations, could already be observed histologically in the fetal stage with the disappearance of embryonal blood vessels. These structures begin in the vitreous base, go into the vitreous, run parallel to the retina, and then go to the back of the vitreous and towards the lens. The physiological mats of vitreal fibril concentrations (membranelles or tractus) and the pathological branches of the persistent hyaloid artery, topographically correspond to the mats of the obliterated embryonal blood vessels of the vitreous. These mats grow in relation to the bulbous growth. CONCLUSIONS: In these investigations an attempt has been made to clarify the question of which embryonal blood vessels. which embryonal and fetal lengths, which different physiological tractus of the vitreous body and which different pathological features of the persistent hyaloid artery correspond. PMID- 8720721 TI - The mydriatic effect of topical glycopyrrolate. AB - BACKGROUND: Glycopyrrolate, an anticholinergic agent that does not cross the blood-brain barrier, has several indications, but its mydriatic effect has never been tested. This study was carried out in order to compare the mydriatic effect of glycopyrrolate 0.5% to that of atropine sulfate 1%. METHODS: Glycopyrrolate 0.5% and atropine 1.0% were instilled separately in the eyes of albino rabbits. Pupil diameter and intra-ocular pressure were monitored. RESULTS: Mydriasis was noted within 5 min of glycopyrrolate instillation, reached near-maximal level at 15 min and persisted for 1 week. Glycopyrrolate 0.5% showed a faster, stronger and more persistent mydriatic effect than atropine 1.0%. Administration of glycopyrrolate 0.5% solution b.i.d. for 1 week did not affect intra-ocular pressure or produce any adverse reaction. CONCLUSION: Glycopyrrolate solution has the potential to deliver an ocular anticholinergic effect without causing associated central anticholinergic hazards. PMID- 8720722 TI - Adult-onset foveomacular vitelliform dystrophy and indocyanine green videoangiography. AB - BACKGROUND: Adult-onset foveomacular vitelliform dystrophy (AOFVD) represents a heterogeneous group of disorders with different clinical, angiographic, and histopathological features. The most common form is characterized by a yellow, round to oval subretinal macular lesion with or without central pigmented spot. METHODS: Eight patients affected by typical AOFVD underwent fluorescein angiography and indocyanine green videoangiography (ICGV). RESULTS: Fluorescein angiography showed a central hypofluorescent spot surrounded by an irregular hyperfluorescent ring. ICGV demonstrated a foveal nonfluorescent spot, visible during the entire examination, and a hyperfluorescent area surrounding the central spot, which became evident soon after the beginning of the examination. CONCLUSIONS: In light of previous histopathological studies, the central nonfluorescent spot may be interpreted as a masking effect of a pigment clump, whereas the hyper-fluorescent area may represent dye pooling or staining of the subretinal pigment epithelial material. PMID- 8720723 TI - [Autologous transplantation of hematopoietic precursor cells following CD34 selection]. AB - Peripheral blood is increasingly used instead of bone marrow as a source of hemopoietic precursor cells for transplantation. The optimal technique still needs to be defined. Selection of CD34+ cells in transplant material may be of benefit in allogeneic and autologous peripheral blood precursor cell transplantation (PBPCT), since it allows elimination of unwanted CD34-negative cells, such as T-cells and contaminating tumor cells. We have evaluated the feasibility of CD34 selection in PB transplants and studied hemopoietic reconstitution after autologous transplantation of CD34 selected precursor cells. Between August 1994 and June 1995 CD34 selection was performed on 12 transplants for 9 patients with malignant disease (non-Hodgkin lymphoma [n = 5]; Ewing sarcoma [n = 1]; chronic lymphocytic leukemia [n = 1]; breast cancer [n = 1]; multiple myeloma [n = 1]). PBPC were collected with a Fenwall CS 3000 harvester after stimulation with G-CSF. For selection of CD34+ cells the Ceprate LC34 system (CellPro) was used. A median CD34 purity of 73% (range 40-94%) was achieved. The median number of CD34 positive cells per transplant was 4.8 x 10(6)/kg body weight (range 0.7-15.8). The median number of colony forming cells per transplant was 31 x 10(4)/kg body weight (range 1.5-131.3). For autologous PBPCT the minimal number of CD34 positive cells required in the transplantate was arbitrarily set at 1.0 x 10(6)/kg body weight. This number was achieved in 10 of the 12 transplants. The median loss of CD34+ cells during selection was 1.5 x 10(6)/kg body weight (range 0.2-6.4). In 2 patients the total number was reduced to below the critical value of 1.0 x 10(6)/kg. 7 of the 9 patients received the CD34 selected transplant after intensive chemotherapy and irradiation. The median follow-up time after PBPCT was 196 days (range 62-278). All 7 patients are now alive and with normal hemopoietic function. A granulocyte count above 0.5 x 10(9)/l and a platelet count above 20 x 10(9)/l was achieved on day 14 (median), and on day 19 after PBPCT. We conclude that CD34 selection is technically feasible and that CD34 selected cells can be used for PBPCT. The procedure is time consuming and expensive; it requires complex organization at laboratory level, and the benefit of CD34 selection with regard to T-cell depletion and tumor purging still needs to be proven. However, CD34+ selection is likely to open new perspectives in transplantation medicine. PMID- 8720724 TI - [Detection of Bartonella (Rochalimaea) henselae/B. quintana by polymerase chain reaction (PCR)]. AB - Bartonella (Rochalimaea) henselae and/or B. quintana are the causative agents of a variety of infections such as trench fever, bacillary angiomatosis, septicemia, peliosis hepatis and endocarditis. Recently, B. henselae has been identified as a major cause of cat scratch disease. Diagnosis of such infections is based on clinical information, histopathology, culture and serology. However, none of these methods alone is sufficiently sensitive or specific. We have used the PCR to search for DNA specific for B. henselae/B. quintana in 33 clinical samples and in 6 controls. In comparison with clinical data and histopathology, PCR was extremely specific (100%) and reasonably sensitive (61%). Possible explanations for the limited sensitivity of PCR are discussed. We conclude that PCR provides a useful adjunct for the diagnosis of infections caused by B. henselae and B. quintana. PMID- 8720725 TI - [Acute vestibular syndrome in cerebellar infarct of the posterior inferior cerebellar artery (PICA infarct)]. AB - The differential diagnosis of an acute peripheral vestibulopathy ("vestibular neuritis") and of an ischemic lesion in the cerebellar territory of the posterior inferior cerebellar artery (PICA) is important. Both present with acute vertigo, vomiting, spontaneous nystagmus and difficulties in walking. We analyze the clinical, oculographic and NMR findings as well as the outcome in 10 patients with an ischemic stroke in the cerebellar territory of the PICA. On clinical grounds alone, it is not always possible to attribute the acute vestibular syndrome to a peripheral vestibular lesion or to a PICA insult. In our experience an important feature of a PICA insult is a discrepancy between the amount of vertigo, the severity of the walking difficulties, and the often weak and rapidly resolving spontaneous nystagmus. In contrast to a peripheral vestibulopathy, the vestibuloocular reflex is only minimally or not decreased in PICA insults, and gain asymmetries are accounted for by the presence of spontaneous nystagmus. However, only neuroimaging can finally differentiate between the two entities. PMID- 8720726 TI - [Spinal stenosis: current aspects of imaging diagnosis and therapy]. AB - Lumbar spinal stenosis typically manifests itself through low back pain and neurogenic claudication during walking. Cervical spinal stenosis may manifest itself through a multitude of symptoms. Spinal stenosis is classified into congenital or primary and acquired or secondary forms. Conventional X-rays of the spine often reveal only unspecific findings. Computed tomography (CT) is the preferred method of judging the width and anatomy of the spinal canal. Myelography remains a good method of imaging the spinal cord in its entirety and in addition allows functional imaging of the width of the spinal canal with functional stress images. CT performed directly after myelography (myelo-CT) allows specific imaging of areas still unclear. Magnetic resonance imaging (MRI) allows multiplanar imaging of the spine and better soft tissue definition. Surgical therapy is discussed in relation to the clinical and neuroradiological findings. PMID- 8720727 TI - [PCR: basics and new developments]. AB - Since its discovery 10 years ago PCR has been introduced for a variety of practical applications. PCR has opened new dimensions particularly in laboratory diagnostics because of its sensitivity, accuracy and speed. In spite of the availability of user friendly kits, basic knowledge is of great importance for the user especially if PCR has to be optimized for special needs or when specific problems arise. The general mechanism of the reaction and the significance of the reaction components and the PCR conditions are discussed initially. Several recent developments in PCR (new enzymes, RNA-PCR, improvements of the specificity, prevention of contamination and development of new equipment) that are critical for the user are shortly introduced. Finally, "long PCR" is discussed in order to demonstrate that even 10 years after the invention of PCR significant new breakthroughs in the PCR technology are still possible. PMID- 8720728 TI - [Use of polymerase chain reaction to reduce the use of animal studies: a project for the expression of the structural proteins of tick-borne meningoencephalitis virus]. AB - Tick-borne encephalitis virus is difficult to propagate because with consecutive passages in cell culture the virus titer will decrease. Stockvirus has to be propagated in young mice. Therefore, every production of virus for research, diagnostic assays or vaccination demands the use of laboratory animals. We decided to clone the part of the viral genome which codes for the structural proteins, and to produce the structural proteins in a suitable expression system. Using reverse transcription, followed by the polymerase chain reaction, we amplified exactly this part of the viral genome, added restriction sites for cloning and a stop-codon. Cloning of this DNA-fragment and expression of the structural proteins of tick-borne encephalitis virus in the baculovirus expression system has thus been possible. Replacement of traditional viral antigen by these recombinant proteins may reduce the need for laboratory animals. PMID- 8720729 TI - [PCR: a look behind the scenes at bovine viral diarrhea virus]. AB - As indicated by their nucleotide sequences bovine viral diarrhea viruses (BVD virus) are genetically quite heterogenous. Despite differences in the nucleotide sequence in the NS3 region, we found that the derived amino acid sequence of all viral strains analyzed was identical and differed from that of classical swine fever viruses, thus allowing differentiation between bovine and porcine pestiviruses. Remarkably, the BVD viral strains were isolated over a time period of some 40 years in Europe, North America and New Zealand. The nucleotide sequence of the 5' untranslated genome region of a BVD virus recently isolated in Switzerland from a calf with hemorrhagic syndrome differs from that of BVD viruses causing similar symptoms in a large epidemic outbreak in North America. PMID- 8720730 TI - -Entrance of PCR for studying murine coronaviruses-. AB - This paper describes a number of applications of the Polymerase Chain Reaction (PCR) in the study of enterotropic murine coronaviruses [mouse hepatitis virus (MHV)]. A diagnostic PCR was developed which detected all of 11 different MHV strains. This fast and reliable method was also able to differentiate MHV from other non-murine coronaviruses. On the basis of this assay a quantitative PCR was designed using a mutant template containing a point mutation which competed for the PCR primers. The amplification and cloning of the structural protein genes of enterotropic MHV strains it plasmid vectors for subsequent sequencing is described. In addition an RT PCR was developed which was able to selectivity detect artificially generated recombinant coronavirus. PMID- 8720731 TI - [Detection of feline coronavirus using RT-PCR: basis for the study of the pathogenesis of feline infectious peritonitis (FIP)]. AB - The aim of this study was to further investigate the pathogenesis and epidemiology of feline coronavirus (FCoV)-infections and among others to determine the prognostic value of a positive result in the RT-PCR for FCoV in serum samples collected from cats with abdominal signs. Viral RNA was isolated from 100 microl of serum and subsequently amplified by a nested RT-PCR using primers binding to a highly conserved region of the 3'-end of the FCoV-genome. Sixty-three serum samples collected from 62 cats with abdominal signs were examined by RT-PCR and the clinical outcome was followed up. Four of these cats with a positive PCR-result are healthy more than 70 months after the collection of the blood sample. It can be concluded that viremia with FCoV does not necessarily lead to FIP and death. With respect to diagnosing FIP, a positive FCoV-RT-PCR is of low prognostic and diagnostic value. It can not be recommended to use this assay as sole indication to euthanize cats. Further studies will have to be carried out to demonstrate if the prognostic and diagnostic value of this PCR-assay in other samples such as peripheral blood mononuclear cells is more reliable. However, this method was found to be an important tool to further study the pathogenesis and epidemiology of FIP. PMID- 8720732 TI - [Identification and characterization of new and unknown coronaviruses using RT PCR and degenerate primers]. AB - A modified method of the reverse transcription followed by polymerase chain reaction (RT-PCR) was developed in order to examine the genome of a recently discovered virus, the porcine epidemic diarrhoea virus (PEDV), which resembled morphologically the coronaviruses. The published sequences of the genomes of various coronaviruses were compared. On the level of the amino acid sequence, conserved regions, common to all coronaviruses, were found in the gene encoding the nonstructural protein 1b as well as in the genes coding for the major structural proteins (S, M, and N). Due to the degeneration of the genetic code, some amino acids may be encoded by different nucleotide triplets. In order to compensate for this degeneration, mixtures of primers were synthesized, containing a variety of nucleotide sequences which together represented all possible codons for the conserved amino acid sequences. This method allowed to amplify and clone approximately 4000 base pairs of the genome of PEDV. An analysis of the genomic sequences revealed that PEDV holds an interesting intermediate position between human coronavirus 229E and Transmissible Gastroenteritis virus. We postulate that the method presented in this contribution may be useful to study and characterize other unknown viruses, especially viruses for which no cell cultures for propagation are available. PMID- 8720733 TI - [Quantification of proviral FIV DNA using competitive PCR]. AB - Quantification of provirus copies is important in the context of different biological questions. The most reliable approach for DNA quantification is a PCR based on coamplification of two templates of similar length, the target sequence and the reference template, sharing the same primer recognition sequences. During the amplification, the two templates compete for the same primer set (competitive PCR, or cPCR) AND consequently amplify at the same rate independently of the number of cycles. The amplified products can be distingushed by their different lengths. After densitometrical analysis, the proviral copy number of experimentally feline immunodeficiency virus infected cats could be calculated, since a known amount of reference template was used. The method described here proved to be very sensitive (10 copies for the competitor-DNA) and was used to quantitate the proviral load during several experiments in which the influence of periodical immunestimulations and the effect of vaccines on the virus load was studied. PMID- 8720734 TI - -PCR for the detection of lentiviruses from small ruminants-. AB - The polymerase chain reaction has all attributes of a promising diagnostic technique. It is rapid, simple to perform and extremely sensitive. In a PCR developed for the detection of small ruminant lentiviruses (SRLV), we found under ideal conditions a detection on sensitivity up to less than 10 template DNA copies. The diagnostic application of PCR was not fully satisfying, even when the technique was refined by the use of a panel of suitable primer pairs. The reliability of PCR in blood and milk samples was much lower than that of antibody detection using ELISA. Interestingly, a positive PCR result was also recorded in 50% of the samples of seronegative animals. Seronegative lentivirus carriers due to delayed seroconversion have been described previously (Rimstad et al., 1993). Due to sporadic occurrence of false positive reactions in spite of contamination control, this result must be interpreted with caution unless the specificity of the fragments can be confirmed by sequencing. Using published sequences of SRLV, we show that sequencing of PCR products, followed by phylogenetic analysis should allow to study molecular epidemiology of field strains. PMID- 8720735 TI - [RT-PCR: a tool for the generation of clones for research on canine distemper]. AB - In the present work an adapted method of the reverse transcriptase polymerase chain reaction (RT-PCR) is described. This procedure was an important tool for cloning a variety of gene sequences derived from oligodendrocytes and canine distemper virus (CDV). Among other applications, the obtained clones were used to produce probes for in situ hybridization and to sequence the inserted DNAs. Due to these molecularbiological techniques new insights were gained into the pathogenesis of CDV-induced demyelination and CDV-persistence in the central nervous system. PMID- 8720736 TI - [Effect of Sarcocystis gigantea extract (SGE) on the replication of human immunodeficiency virus (HIV)]. AB - Basis of this study were the previous findings regarding isolation and characterization of a Sarcocystis gigantea lectin (SGL) especially the activation of human mononuclear cells (CD3+, CD4+, CD8+, Mo, B-lymphocytes). HIV susceptible, immortaliced cell lines (H9-, MT-4) should be investigated to examine their reactivity against SGE which contains this strong mitogen. Using lymphocyte proliferation assay a strong stimulation of noninfected CD(4+) positive H9-cell by SGE was observed. HIV-infected H(9+)-cells showed after SGE stimulation (20-160 micrograms) an exacerbation with an optimum at day 4. The virus replication in the H(9+)-hostcells was 13 times stronger. This result could also be detected indirectly because of the higher cytotoxicity in the MT-4 cell system. Cytopathogeny was measured by MTT cellvitality assay. In parallel, the high sensitive p24 core Profile-ELISA was used to directly examine the amount of produced HIV. In case of the H(9+)-cells the virus release per cell was 5 times higher after SGE stimulation compared with control. PMID- 8720737 TI - [Determination of concentration and emission of gases relevant to animal and environmental hygiene in different systems for fattening of pigs with the aid of multigas monitoring]. AB - Multigasmonitoring is a method to measure continuously gases relevant from the view point of animal and environmental hygiene with high sampling rate and to calculate emission. The methodical procedure has been described by example from different deep litter systems with additives and slatted floor system with liquid manure storage for fattening pigs. It has been shown that the emission of ammonia from an obviously good working deep litter system can be lowered by approximately 30% in comparison with slatted floor. The mean concentration of nitrous oxide in deep litter system with additives was approximately 2 ppm, whereas in the liquid manure system the N2O concentration was less than the outdoor value. PMID- 8720738 TI - [On calving in the pasture during the fall (veterinary expert)]. AB - The objective of the following study was a veterinary expert opinion dealing with the evaluation of the pasturing of a cow and her calf during fall with special respect to animal welfare. The legal basis of this expert opinion was the Animal Welfare Act as amended on 1993. It is emphasized that there are no binding legal regulations as to the pasturing of livestock and that, especially because of the increasing public interest in animal welfare matters, the definition of minimal requirements would be desirable. PMID- 8720739 TI - [Animal welfare aspects of the production of veal]. AB - White veal calf fattening by reducing iron in feed results in clinically disturbed state of health, anaemia, loss of weight and inferior growth of calves. Natural variability of meat quality allows to differentiate in young normally fed calves at the age of 2 till 5 weeks between those with either more anaemic white or more natural rose-coloured veal. So now the consumer of white meat can meet his preferred wishes. The parameters of meat: water percentage, water loosely bound, protein, fat, collagene, elastine, ferrum, copper and zinc don't differ in both variants of quality. The same applies to the minerals in the spleen and also to ferrum in the liver on the one hand. On the other hand copper and zinc really differ in the livers of anaemic versus those of calves adequately fed. PMID- 8720740 TI - [From animal automatons to scientific questioning--reflections on the development of veterinary medicine in the 18th century]. AB - Descartes wanted to prove the cosmos to be a machine which God had made run. The animals, too, were machines which functioned like robots. In the 18th century the animal machine of Descartes was being discussed and eventually negated. The animal was considered a being with a soul and an intellect. Therefore an animal was to be looked at as a creation of God-nearly like a human being. Reimarus (About the animals' soul, 1760) and Germershausen (About sheep breeding, 1789), tried to be fair to animals while handling them. They negated the machine model of Descartes but accepted his scientific language. They transferred his vocabulary into their treatises. In this way they installed this new vocabulary into their scientific terminology--an important requirement for the instutitionalisation of veterinary medicine. PMID- 8720741 TI - Low dose corticosteroids in early rheumatoid arthritis. Can these drugs slow disease progression? AB - The role of corticosteroids in treating rheumatoid arthritis is controversial, but recourse to the available evidence of efficacy should guide patient management decisions. Earlier evidence suggested that symptomatic control could be improved for periods of 6 to 12 months, but not longer, without increasing doses to unacceptably high levels. The effect of corticosteroids on joint destruction has been unclear. Recent findings from a controlled clinical trial show that prednisolone 7.5 mg/day can significantly retard the rate of erosive progression in patients with relatively early disease (< 2 years' duration). These results have implications for both disease management and our understanding of the pathogenesis of joint destruction in rheumatoid arthritis. PMID- 8720742 TI - Antimicrobial resistance patterns in long term geriatric care. Implications for drug therapy. AB - There is a high prevalence of bacterial infections in long term care facilities (4.4 to 16.2%). This, together with the fact that antimicrobial resistance is a big concern in current medical practice, makes infection control so important in nursing home care. This article covers the mechanisms of antibacterial resistance and focuses on 4 major antibacterial-resistant bacteria. Vancomycin is the treatment of choice for methicillin-resistant Staphylococcus aureus (MRSA). Colonisation with MRSA is not uncommon in nursing homes and eradication is probably not necessary. Any clinically important enterococcal infection should be tested for high-level resistance. An infectious disease consultation should be sought for vancomycin-resistant enterococcal infections. Gram-negative bacilli have developed multi-resistance. Susceptibility testing can identify the most appropriate therapy. Multiresistance should also be considered when treating Streptococcus pneumoniae. Overall, handwashing is highly recommended. Barrier precautions, minimising hospitalisations and avoiding unnecessary personnel rotation can reduce the chance of resistance spread. PMID- 8720743 TI - Pharmacological properties of hirudin and its derivatives. Potential clinical advantages over heparin. AB - Hirudin and its derivatives represent the first parenteral anticoagulants introduced since the discovery of heparin in the early 1900s. Hirudin, the naturally occurring anticoagulant of the leech, is a single peptide chain of 65 amino acids with a molecular weight of about 7000. Recombinant technology has developed methods to produce recombinant forms of hirudin (r-hirudin) in sufficient quantities for therapeutic use. Hirudin is a potent thrombin-specific inhibitor that forms equimolar complexes with thrombin. It represents a new anticoagulant agent in a field in which heparin has been the only available drug for many years. In contrast to heparin, hirudin does not require antithrombin III as a cofactor, is not inactivated by antiheparin proteins, has no direct effects on platelets and may also inactivate thrombin bound to clot or the subendothelium. In humans, experience with r-hirudin in preventing or treating venous thromboembolism is very preliminary. However, r-hirudin achieved promising results in patients with unstable angina, or following coronary angioplasty. In patients with acute myocardial infarction, 3 important clinical trials were stopped because of an excess of bleeding complications. At present, the discovery of a r-hirudin regimen that is more efficacious than heparin and at least as safe needs a reappraisal of the drug in further trials. PMID- 8720744 TI - Insulin treatment in elderly patients with non-insulin-dependent diabetes mellitus. A double-edged sword? AB - Elderly patients with non-insulin-dependent (type 2) diabetes mellitus (NIDDM) form one of the largest sectors of the diabetic population. Emerging evidence indicates that hyperglycaemia is associated not just with an increased risk of microvascular complications but also with macrovascular disease, which remains the main cause of excess mortality in people with NIDDM. The treatment of hyperglycaemia in patients with NIDDM is notoriously difficult when diet, exercise and judicious use of oral antihyperglycaemic agents fail to maintain acceptable metabolic control. The treatment of hyperglycaemia in elderly patients is further hampered by age- or disease-related comorbidity. Insulin therapy can ameliorate many metabolic abnormalities of NIDDM, with consequent reduction of hyperglycaemia. Moreover, insulin treatment induces antiatherogenic changes in serum lipids and lipoproteins and probably enhances general well-being. However, insulin therapy is associated with bodyweight gain and an increased risk of hypoglycaemia. An unresolved question is the relationship of exogenous insulin therapy to the development of cardiovascular diseases. This reverse side of the coin has prompted research aimed at establishing methods to achieve the best possible reduction in hyperglycaemia with the smallest dose of insulin as possible. The most promising target in this respect has been the control of glucose overproduction from the liver by the nocturnal administration of intermediate- or long-acting insulin with or without oral antihyperglycaemic drugs. Intensive insulin therapy does not seem to have clear-cut benefits in elderly patients and can be hazardous. However, we cannot at present predict who will benefit from the various therapeutic regimens and therefore clinicians should use sound clinical judgment in choosing the appropriate therapy for an individual patient with NIDDM. Although we do not know at present whether we can, by our current modes of treatment, lower the frequency of vascular diseases, therapeutic nihilism, even in elderly patients with NIDDM, is outmoded. PMID- 8720747 TI - Health education and the prevention of coronary heart disease. PMID- 8720748 TI - Myelodysplastic syndromes. PMID- 8720745 TI - Hormone replacement therapy in the aged. A state of the art review. AB - The use of hormone replacement therapy (HRT) in the immediate postmenopause for the relief of menopausal symptoms and for the prevention of osteoporosis and cardiovascular disease is well established. The continuation of treatment beyond the age of 60 years is likely to maximise these long term benefits and there is now increasing evidence to suggest that commencing treatment de novo in women of this age is likely to be beneficial. Many women remain symptomatic well into their sixties and the introduction of HRT at this stage will not only relieve these symptoms but will also conserve bone density and reduce future osteoporotic fracture risk. Furthermore, HRT appears to reduce the risk of cardiovascular disease, even in those women with pre-existing heart disease. The possible association between HRT and breast cancer remains controversial. Overall, there seems to be a slight increase in risk with long term HRT usage (longer than 10 years) but certain subgroups of women may be more at risk. This review discusses the merits and potential problems of prescribing HRT to the elderly and gives some guidance on the type, dose and route of administration of estrogen and progestogen to be used. Poor compliance with HRT is a major problem and the more widespread use of pretreatment counselling together with a wider range of products should have a positive impact in this area. The final decision about whether to continue or commence HRT in the elderly should be an informed one made by the woman and her clinician together. PMID- 8720746 TI - Ticlopidine. A review of its pharmacology, clinical efficacy and tolerability in the prevention of cerebral ischaemia and stroke. AB - Ticlopidine is a thienopyridine derivative which reduces the risk of reversible ischaemia and stroke in patients who have previously experienced a cerebral ischaemic episode. In comparison with aspirin, ticlopidine produced a significant reduction in the risk of stroke in a multicentre clinical trial involving more than 3000 patients with previous transient or persistent minor ischaemia, and was superior to placebo for the prevention of stroke recurrence in more than 1000 patients who had experienced a major thrombotic stroke. The cost-utility ratio for ticlopidine in comparison with aspirin was estimated to be $US31 200 to 55,000 per quality-adjusted life-year gained. Diarrhoea is the most common adverse event in ticlopidine recipients (20 to 22% incidence versus about 10% with placebo), although skin rash, nausea, dyspepsia, bleeding events, abnormal liver function and haematological disturbances were also observed in clinical trials. Severe neutropenia is the most serious event: this developed in 0.85% of patients receiving ticlopidine in 2 large clinical studies (n = 4098) but resolved after treatment withdrawal. Fatal neutropenia, although rare, has been reported in some patients receiving ticlopidine. Thus, ticlopidine is effective in reducing the risk of recurrent cerebral ischaemia and stroke. It appears to provide a gain over aspirin for the prevention of stroke after reversible ischaemia, particularly during the first year of treatment (when the risk of stroke is greatest), although further data on its absolute relative benefit would be useful. The extent to which ticlopidine is prescribed will probably depend on individual clinicians' perception of its risk/benefit and cost-effectiveness profiles. Ticlopidine is likely to be particularly useful for stroke prophylaxis in patients who do not tolerate aspirin or who have an ischaemic episode during aspirin treatment, and for the prevention of stroke recurrence in patients who have previously experienced a major stroke. PMID- 8720749 TI - Heart transplantation: initial results in Scotland. AB - Between 1st Jan 1992 and 1st Sept 1993 164 patients were referred as candidates for heart transplantation in Scotland. 79 were accepted for transplantation and 39 of these received orthotopic heart transplants. 30 day mortality was zero. Mean time on intensive care post operatively was 46.35 hrs and mean time to hospital discharge was 17.03 days. Cumulative actuarial survival (survival at 18 months post operatively) was 87%. There were three deaths, two from infection at six and 19 weeks and one from neoplasia at 37 weeks. Mean time to rejection was 38.54 days. Linear rejection rates at 0-3, 3-6, 6-9, 9-12, 12-15 and 15-18 months were 1.86, 0.94, 0.6, 0.04, 0 and 0 events per 100 pt-days respectively. Linear infection rates at 0-3, 3-6, 6-9, 9-12, 12-15 and 15-18 months were 1.81, 0.17, 0.1, 0, 0.05 and 0.18 events per 100 pt-days respectively. The heart transplant programme in Scotland has been successful in terms of operative mortality and short to medium term survival. PMID- 8720750 TI - Impact of a health promotion programme on multiple risk factors for CHD: a preliminary evaluation. AB - Results are reported from a pilot study to evaluate an innovative health promotion programme (Lanark Health Club), in a community where the incidence of CHD is above average. The aim of the programme is to provide multiple risk factor screening and interventions--including an exercise programme--sited in the community and utilising existing resources within primary care and the voluntary sector. Data were returned by 54 respondents after six months' participation on the programme. Subjects' self-reported activity levels increased significantly over time, and there were concomitant significant reductions in weight, SBP and DBP. Reduced levels of self-reported stress were associated with reductions in alcohol intake, and were greater in male subjects. When other measures were controlled for, stress reduction was significantly more likely for those who also lost weight and exhibited a reduction in DBP. Finally, participation in the Health Club appeared to result in cholesterol reduction, particularly for those with relatively high initial cholesterol levels. While this could reflect regression effects, it is certainly worthy of more systematic examination. PMID- 8720751 TI - Multiple pancreatic abscesses due to Candida albicans following ERCP. AB - A report is presented of a patient who developed multiple abscesses of the pancreas due to Candida albicans following an Endoscopic retrograde chole pancreatography (ERCP) for acute pancreatitis. He was not immunocompromised, debilitated and had not had recent surgery. There was complete radiological and clinical resolution of the abscess on prolonged treatment with amphotericin alone. Only a few cases of candidal abscess of the pancreas have been reported, none of them having occurred after an ERCP. PMID- 8720752 TI - Tuberous sclerosis--an unusual cause of seizures in an 18 year old. AB - An 18 year old man presenting with seizures was found to have hypomelanotic macules and a cardiac rhabdomyoma. These features suggested a diagnosis of tuberous sclerosis (TS) but there were no other clinical signs, no family history and cranial imaging failed to reveal the characteristic appearances. The diagnostic criteria for TS are reviewed and the importance of thorough clinical examination and appropriate investigation in this disease is stressed. PMID- 8720753 TI - The changing role of an audit support group. AB - An audit support group was established within a University Department of Public Health in 1991 to provide a resource for clinical audit. A postal survey of 300 consultants and senior registrars (59% response rate) showed 72% had participated in audit, 47% currently involved, 40% implementing change. 40% had no experience in standard setting or implementing change. Comparison of the workload of the group for August 91-March 92 and April 92-March 93 showed 4 requests for support per month, 4/5 from doctors during first period compared with 6 requests per month and 1/3 from doctors during second period. Since the group was established demand from clinicians has decreased and from professions allied to medicine increased. This may reflect either clinicians' increased experience or decline in enthusiasm for audit. Education in the basic skills for audit for all healthcare professionals, improved communications and practical support for clinical audit is necessary. PMID- 8720755 TI - In Larrey's shadow: transport of British sick and wounded in the Napoleonic wars. AB - Dominique Jean Larrey's introduction of a 'flying ambulance' into Napoleons army was a great advance in military surgery. British arrangements for the transport of wounded and sick during the Napoleonic wars fell far short of Larrey's humanitarian vision. The efforts of eminent doctors such as Sir James McGrigor to create a formal ambulance service were frustrated by the army establishment. As a result the sick were often abandoned or carried in local bullock carts. Only sixty years after Waterloo did Britain finally follow Larrey's example and form a field organisation including a trained ambulance corps. PMID- 8720754 TI - An audit of utilisation of Holter tape facilities at Glasgow Royal Infirmary. AB - Holter tape analysis is widely used to both confirm and exclude the presence of significant arrhythmias. We have studied the utilisation of the Holter service at Glasgow Royal Infirmary over a period of 6 months. During this period 305 x 24 hour Holter tapes were recorded. Out patient referrals generated 95% of these requests with only 5% of requests for in patient recordings. The median delay from request date to dispatch to the referring doctor was 14 days. No arrhythmia was detected in 77% and only 3% had a detectable symptomatic significant arrhythmia. The recording was felt to demonstrate the presence of a relevant arrhythmia in 17% of cases and to help exclude a possible arrhythmia in 61%. The capital cost per tape was 20.82 Pounds. In conclusion Holter monitoring is an inexpensive way of analysing the heart rhythm over an extended period which is probably underused. PMID- 8720756 TI - Left ventricular hypertrophy in hypertension--general and local trends in diagnosis and therapy. AB - LVH is a frequent echocardiographic finding in the general population but it should be regarded as an ominous predictor of future cardiovascular events rather than an innocent compensatory phenomenon. Echocardiography is the most sensitive and specific method for its detection but the ECG should not be regarded as superfluous since additional prognostic information and information about coexisting myocardial damage is present. The decreasing prevalence of LVH suggests that LVH can be prevented by control of hypertension and prevention of weight gain. Once LVH is present then antihypertensive drugs, weight reduction or salt restriction may promote its reversal, with early studies demonstrating associated improvement in mortality and morbidity. PMID- 8720757 TI - The importance of being systematic. PMID- 8720758 TI - The impact of travel acquired illness on the world traveller and family doctor and the need for pre-travel health education. AB - This retrospective study involved a 20% quota, age and sex stratified sample of people over 16 years of age, presenting to a group medical practice over a period of one year (N = 1568). A standardised, computer-scored, self-report questionnaire was administered. The response rate was 98.3%. 42% of respondents had travelled outwith the UK in the previous year. Of those, 42% had become ill whilst abroad. In 20.5% of cases the illness settled without treatment. However, 26% of the patients required consultation with a doctor whilst abroad and 48.4% of those becoming ill required further attention from the family doctor on return home. 5% of ill travellers were admitted to hospital abroad. 8% of all travellers did not have medical insurance cover. A large and significant population of travellers become ill whilst abroad, and travel-acquired illness has a large impact on general practice, with 1 in 5 travellers seeking GP consultation on return home. Improved practice-generated pre-travel health advice might decrease this burden on primary care. PMID- 8720759 TI - The management of operable breast cancer in Scotland surgeons' opinions. The Scottish Cancer Trials Breast Group. AB - This report gives the responses of general surgeons in Scotland to two questionnaires. Satisfactory rates were obtained: 82% for the more detailed survey in 1988 and 62% for the second survey (1991), where nonrespondents were not followed up. In 1988 the rationale was the poor participation in the Scottish breast conservation trial while the 1991 survey further investigated the diversity of surgical opinion identified in the first. The limited trial support in 1988 was mainly due to reluctance to accept all treatment options. The majority were prepared to consider trial participation although 47% believed this compromised doctor-patient relationships. Although breast-conserving therapy was widely supported, many different views on management were encountered, both in the degree of influence of specific factors and in the decisions taken in relation to them. We believe these surveys have re-inforced a need for management guidelines, particularly as around half the patients with symptomatic breast cancer were being treated in non-specialist units. PMID- 8720760 TI - Systemic hypersensitivity reaction to intravesical BCG. AB - BCGosis is a recognised complication of intravesical BCG immunotherapy for superficial bladder carcinoma. We present here a case of granulomatous hepatitis, pneumonitis and probable pericarditis resulting from an hypersensitivity reaction to intravesical BCG. PMID- 8720761 TI - The early hospital readmission of elderly people. AB - Two hundred and twenty six consecutive patients aged over 75 years discharged following an acute medical admission to a district general hospital were followed up six months later. 46 (20.3%) required emergency admission within three months of discharge. 19 had a diagnosis related to their original admission, and 27 an unrelated diagnosis. The mean time to readmission was 34 days, but this was significantly shorter (21 days) in those with a related diagnosis. Those readmitted had more often required admission in the year prior to their index admission, had more comorbidities, higher use of social services, and no carer. There were no significant differences in age, gender, mental test score, length of stay, home circumstances or diagnosis. The results suggest that most readmissions are due to medical rather than social problems, and that most are unavoidable. Efforts to reduce readmissions should be directed at preventing early medical relapse in at risk patients. PMID- 8720762 TI - A social and personal history of William Harvey 1578-1657. PMID- 8720763 TI - Community care and the acute services. PMID- 8720764 TI - Community care--one step forward or two steps back? PMID- 8720765 TI - Community care one year on. PMID- 8720766 TI - Antihypertensive drugs and glucose metabolism: a comparison between a diuretic and felodipine, a new calcium antagonist, when added to a beta-blocker in non diabetic hypertensive women. AB - Felodipine, a vascular selective antihypertensive calcium antagonist, was compared with hydrochlorothiazide, a diuretic, with respect to glucose tolerance. An open crossover study was performed comprising 16 non-diabetic hypertensive women (age range 59-75 years). The women continued to take a beta-blocker as a basal therapy. Each treatment period lasted three months. The blood pressure was similar irrespective of treatment. Blood glucose values were not significantly different during the oral glucose tolerance test. Serum insulin levels after glucose administration were lower when the patients were treated with felodipine than when taking hydrochlorothiazide. A possible explanation for this observation may be an increased insulin release as a consequence of treatment with a diuretic in order to maintain normal blood glucose levels during the glucose tolerance test. Felodipine appears preferable to hydrochlorothiazide as an addition to a beta blocker in hypertensive patients from a glucose metabolism point of view. PMID- 8720767 TI - The prevalence of smoking amongst patients attending an out-patient lipid clinic. AB - The smoking habits of 184 attendees (mean age 47 years, 101 men) at a hospital based lipid clinic were reviewed in conjunction with a measurement of serum thiocyanate level which is a marker for smoking over the previous two weeks. Advice against smoking is routinely given at the clinic. We found 84 (46%) patients to remain active smokers of whom 78 admitted to smoking and 6 were identified on the basis of a serum thiocyanate level greater than 100 micromol/l. Smoking was more prevalent amongst women than men (52% v 33%). We have identified a high proportion of patients attending an outpatient lipid clinic as remaining active smokers despite receiving advice against cigarette smoking. This emphasises the need for more effective counselling against smoking in a multiple risk factor intervention approach to ischaemic heart disease. PMID- 8720768 TI - Deficiencies of management of spontaneous pneumothoraces. AB - In a retrospective, case-note based, audit of the initial management of spontaneous pneumothoraces in a large teaching hospital, 38 events were identified in 37 patients admitted over 12 months to the end of September 1991. The initial management was intercostal tube drainage in 21 (55%), decided upon by middle grade staff, with a cost in terms of the duration of hospitalisation (median 7 (range 2-18) days), chest radiography (6 (3-10) films) and potential for complications including infections (3 wound, 2 chest). Aspiration was performed in only three cases, the remainder were managed by observation. We suggest that undue reliance is placed upon intercostal tube drainage to treat spontaneous pneumothoraces with aspiration alone performed too infrequently. Moreover, other deficiencies in management including the practise of drain clamping and incomplete follow-up were identified. PMID- 8720769 TI - Cardioversion of atrial fibrillation: how important is a long arrhythmia duration? AB - A long duration of atrial fibrillation is usually considered as an adverse feature for successful cardioversion of atrial fibrillation and the maintenance of sinus rhythm. This often leads to the exclusion of such patients from being considered for this procedure. We report three patients in whom atrial fibrillation was present for a long duration (one for 2 years, two for 10 years), and successful cardioversion to sinus rhythm was achieved. Proper selection of patients with atrial fibrillation with an understanding of all features predicting a successful outcome will often allow a good result. PMID- 8720770 TI - Investigation and surgery in irritable bowel syndrome--a cautionary series. AB - Irritable bowel syndrome is a common problem, and, with a broad symptom complex, likely to present to a wide variety of clinicians. Lack of awareness of these manifestations may lead to inappropriate investigation, and surgery. Six patients with irritable bowel syndrome who had multiple investigations and operations are reported. There was a median of seven years delay in diagnosis, patients seeing a median of six different consultants. Over 13 years of follow-up symptoms were unchanged, but patients had undergone a median of eight (often invasive) investigations, and six operations. Several operations were for complications of, or resulted from, a previous procedure. There were far fewer operations after diagnosis, four over 33 patient-years, than prior to diagnosis, 25 over 43 patient-years (chi-squared = 8.074, 1 df, p < 0.0025). All clinicians should be aware of the diffuse symptomatology of irritable bowel, and its potential to mimic other diseases. Surgery should be avoided. PMID- 8720771 TI - Hypothyroidism presenting with cardiac tamponade. AB - Two cases of cardiac tamponade initially suspected to be secondary to malignancy are presented. Primary hypothyroidism can cause pericardial effusions and thyroid function tests confirmed this diagnosis in these cases. Hypothyroidism should be considered as an underlying cause of pericardial effusion in cardiac tamponade. PMID- 8720772 TI - Nail gun injury to the brain: an unusual case of suicide. AB - An unusual case of fatal suicidal craniocerebral penetrating injury due to a nail gun is described. The victim, a 52 year old joiner experienced in the use of nail drivers, shot himself just above the forehead in the midline, driving the nail through his hypothalamus and midbrain. Death was delayed by nearly 24 hours. PMID- 8720773 TI - An audit of sickle cell screening in a paediatric hospital population. AB - Current UK guidelines for the detection of sickle cell disease (SSD) recommend pre-operative screening of patients of African or Afro-Caribbean descent and of patients from the Middle East, South Italy, Greece and India. These recommendations have considerable cost implications for the NHS. We have undertaken a retrospective audit of the use of SSD screening in our own institution. Between January 1987 and August 1992, 1305 patients under 15 years of age (mean 3.8 years) underwent screening for SSD. Surgical patients accounted for 1120 tests (85.8%). The majority of patients tested were of Asian descent (n = 955; 73%); only 350 (27%) were of African or Afro-Caribbean descent. Only four patients (0.3%) tested positive (all sickle cell trait); three of Afro-Caribbean descent and one from Bahrain. We conclude that, in our population, the incidence of HbS is low. Patients of African or Afro-Caribbean descent should continue to be tested but a selective policy may be indicated in other ethnic groups. PMID- 8720774 TI - Alfred Ernest Maylard, 1855-1947: Glasgow surgeon extraordinaire. AB - This account concerns Alfred Ernest Maylard, 1855-1947, a surgeon of exceptional ability and a man with extraordinary energy and drive. He was appointed to the Victoria Infirmary, Glasgow, when it opened in 1890 and made an outstanding clinical and academic contribution, particularly in abdominal surgery. His organisational talents were remarkable and manifest in his influence not just upon the hospital's developments but also upon Glasgow's medical life in general. In his spare time he was a keen mountaineer and was the driving force behind the formation of the Scottish Mountaineering Club in 1889 and its first secretary and later president. He was a prolific writer in all these fields, his eminence as scientific surgeon being recognised by election to the Fellowship of the Royal Society of Edinburgh and his contribution to mountaineering by the vice presidency of the Royal Scottish Geographical Society. PMID- 8720775 TI - [Genes for cellular mortality]. PMID- 8720776 TI - [Cellular senescence, cancer and telomerase]. PMID- 8720777 TI - [Gene expression during replicative senescence]. PMID- 8720778 TI - [Mapping of a senescent gene on human chromosome 7]. PMID- 8720779 TI - [Positional cloning of the genes for progeroid syndromes]. PMID- 8720780 TI - [A variety of halogenating enzymes and their roles in organisms]. PMID- 8720781 TI - [Insulin-like growth factor-I controls animal growth]. PMID- 8720782 TI - [Development of new non-radioisotopic DNA sequencing method by using chemiluminescent DNA detection system]. PMID- 8720783 TI - A clinical anatomy curriculum for the medical student of the 21st century: gross anatomy. AB - Human anatomy forms the foundation for clinical medicine; thus its place in the medical school curriculum deserves careful attention. In an attempt to provide guidance to decision-makers involved in clinical anatomy curriculum development at the medical school level, the Educational Affairs Committee of the American Association of Clinical Anatomists (AACA) developed this document, which defines the contours of a gross anatomy curriculum leading to the M.D. or D.O. degree. The main body of the document sets forth the anatomical concepts as well as the subject matter a student should master prior to graduation from medical school. The AACA seeks to ensure that all medical students receive thorough training in clinical anatomy and that each student, regardless of the institution attended, will be exposed to a curriculum that will provide a fundamental level of competence required for the practice of medicine. PMID- 8720784 TI - Biomechanical alterations in the carpal arch and hand muscles after carpal tunnel release: a further approach toward understanding the function of the flexor retinaculum and the cause of postoperative grip weakness. AB - The difference between maximal and minimal distance covered (the distance between the trapezium ridge and hamate hook; moment exerted on structures: 1 Nm) by an intact flexor retinaculum (FR; minimum, 3.3 +/- 0.1 cm; maximum, 3.7 +/- 0.2 cm) and the increase in the maximal distance on carpal tunnel release (CTR; increase, 1.6 +/- 0.2 mm) were significant. Under an external supination moment, the distance between the attachments of the trapeziopisiform band increased after CTR. Under external pronation and ulnar abduction moments, the distance between the attachments of the scaphoideohamate band increased after CTR. The CTR resulted in an anatomic attachment loss for the following muscles: the superficial head of the flexor pollicis brevis (shortening by approximately 25%, relative to rest length), the ulnar part of the abductor pollicis brevis (with opposition and adductory functions, approximately 20%), the opponens pollicis (approximately 20%), the middle part of the abductor pollicis brevis (approximately 7%), and the opponens digiti minimi (approximately 10%). Preoperative and postoperative (2-7 weeks after surgery) measurements of the reaction force of the distal phalanx (under isometric thumb opposition and finger II-IV flexion with extended carpal joint) led to differentiation of three groups: (1) significant strength loss--the patients showed difficulties with grasping, lifting, twisting off lids and caps, screwing, pulling ropes, and pinching; (2) no significant change in force values; and (3) a significant increase in strength (patients who could grip more firmly). PMID- 8720785 TI - Angle of torsion of the femur and its correlates. AB - Unpaired femora (171), devoid of gross pathology and grouped by gender (94 male and 77 female) and side (88 left and 83 right), were used to measure the angle of femoral torsion and the maximum femur length and to score the degree of prominency of the superior cervical tubercle, intertrochanteric line, quadrate tubercle, linea aspera, and adductor tubercle. The angle of torsion ranged from 9 to +35 degrees with a mean of +12.3 degrees. The means were not significantly different either by gender or side. The angle correlated negatively with superior cervical tubercle, intertrochanteric line, and adductor tubercle (P < 0.001), positively with quadrate tubercle (P < 0.001) but not with linea aspera, neck shaft angle, or length of femur. Bony prominences were significantly more apparent in males. There was no significant association between prominency and side. The torsion seems to be brought about by muscular activity and capsular and ligamentous strain at the hip. This study suggests to clinicians the possibility of correction of torsion defects in certain hip diseases of growing children by suitable alteration in posture of the lower extremity. PMID- 8720786 TI - Severe cardiovascular disease and Alzheimer's disease: senile plaque formation in cortical areas. AB - The main objectives of this study were to analyze the distribution of senile plaques (SP) and neurofibrillary tangles (NFT) in different cortical areas of patients suffering from severe cardiovascular diseases (CVD) and to compare them with Alzheimer's disease (AD) cases. Forty brains were divided into three groups: an AD group (n = 12), a CVD group (n = 17), and a nonheart disease control group (n = 11). The cortical areas examined were the middle frontal gyrus, the superior and inferior watershed areas, the hippocampal formation with the transentorhinal cortex, and the primary visual cortex. SP and NFT were counted in Bielschowsky stained sections from all cortical areas and from the hippocampal formation and the transentorhinal cortex, respectively. Patients with severe CVD occupied an intermediate position in the spectrum of SP formation between AD and nonheart disease patients. The CVD group showed a higher prevalence of SP than the control group, and SP counts were significantly larger in the inferior watershed area, dentate gyrus, subiculum, and transentorhinal cortex. The distribution of SP was similar in CVD and AD patients. Control and CVD patients showed no difference regarding the number of NFT. The existence of a possible cardiovascular component in the genesis of SP is discussed. PMID- 8720787 TI - The superficial ulnar artery: development and surgical significance. AB - The upper limbs of 72 formalin-fixed human cadavers were examined by dissection for arterial anomalies. In one subject, the ulnar artery was noted to be a branch of the second part of the axillary artery on both right and left sides. It ran a superficial course in the arm, crossed the elbow immediately subjacent to the median cubital vein, and continued its course in the forearm in a subcutaneous position. In the hand it played a dominant role in the formation of the superficial palmar arch. The anomalous ulnar artery was of a smaller caliber than both the radial and common interosseous arteries. Although superficial ulnar arteries have been reported in the literature, the combination of bilateral superficial ulnar arteries originating from the axillary arteries appears to be rare. The developmental and surgical significance of the findings are discussed. PMID- 8720788 TI - Congenital anomalies of left renal vein and its clinical importance: a case report and review of literature. AB - Detailed knowledge of the anatomy and anomalies of renal veins is necessary for retroperitoneal surgery and venographic procedures. According to Thomas (1970, Arch. Surg. 100: 738-740), the anomalies of renal veins are more frequent than estimated. The number of surgical procedures and radiologic examinations related to the retroperitoneum are increasing, and therefore pathologic conditions of the retroperitoneal area have been discussed more frequently. We report on a retroaortic left renal vein joining the left common iliac vein, discuss the embryology and clinical importance of renal vein anomalies, and give an overview on the relevant literature. PMID- 8720789 TI - Reaffirming the importance of dissection. PMID- 8720790 TI - [Development and actual status of animal welfare legislation]. AB - The author informs on the status of animal welfare legislation in the EU as well as in the Federal Republic of Germany. PMID- 8720791 TI - [Animals as co-creatures--moral theological reflections on our responsibility to animals]. AB - Moral obligations between men and animals are derived from historical source, mainly from the Bible, using the way of theology. From human obligations to animals does not necessarily follow that animals have rights. PMID- 8720792 TI - [Prospects and limits of animal breeding from the physiological aspects]. AB - Livestock breeding is characterized by substantial increases in performance in recent years. However, pathophysiological changes have to be considered when increased production levels will exceed maximal physiological capacities. Gastrointestinal adaptation processes of ruminants kept under aride grazing conditions are discussed in comparison with european sheep breeds. The high stress susceptibility in pigs which is genetically determined is taken as a further example to explain the onset of diseases on the basis of pathophysiological mechanisms being induced by breeding techniques. From this the necessity for a close cooperation between animal breeding and physiology has to be concluded. PMID- 8720794 TI - [Ten years of the Veterinary Association for Animal Welfare (TVT)]. AB - The Veterinary Association for Animal Welfare was founded in 1985. Within ten specialised working groups scientists, practitioners and veterinary officers prepare answers to actual welfare problems for legislation, administration, and welfare organisations and offer seminars and training programms. PMID- 8720793 TI - [Activity of a veterinarian within the French fire departments (animal welfare, accidents, disasters)]. AB - The report gives a survey of the pertinent situation in France, the legal principles, the way of action, and the tasks of veterinarians engaged in French fire-brigades. PMID- 8720795 TI - [Animal welfare slaughter regulation]. AB - The animal welfare regulation on the slaughter of animals, existing only as a draft for the time being, is designed not only to transpose EC-legislation into national law but also to update and strengthen preconstitutional national legislation on this matter. For a wide area related to the slaughter or killing of animals, animal welfare requirements are put in concrete terms. Among the topics belonging to this area are the theoretical and practical knowledge of the personnel, the handling of animals before slaughter or killing, stunning, the control of its efficacy and the permessibility of certain stunning or killing methods. Not only livestock but also, for example, fur animals and fish are concerned. In practice it will take some efforts in order to attain compliance with the provisions of the animal welfare slaughter regulation. PMID- 8720796 TI - [Applied animal welfare of slaughter houses]. AB - Applied Animal Welfare at Slaughter Houses. The activities of the training- and consultancy institute for careful handling of breeding- and slaughter-animals (bsi) which exists for 3 years now are described. Some major critical points and solutions are discussed. PMID- 8720797 TI - [Mobile slaughter facilities]. AB - Mobile slaughter is commonly discussed as alternative method to slaughter avoiding transportation stress in slaughter animals. In 1994 mobile slaughter became part of the coalition-contract between the two major parties, SPD and GRUNE, in the Department HESSEN. In the article the actual principles, problems to cope with EU-legislation and hygiene-standards, chances and risks of mobile slaughter are discussed. Assuming that some of the major problems concerning hygiene, workers security and waste-deposit can be solved, mobile slaughter seems to bring an improvement in animal welfare and advantages in related meet quality. The slightly higher costs may be acceptable in welfare-interested, high-quality market sections. PMID- 8720798 TI - [Carbon dioxide stunning for the slaughter of turkeys]. AB - In an experimental study on stunning of 460 turkeys in a CO2 enriched atmosphere (60-70% CO2 in air), as a more animal protecting and more economic alternative to conventional electrical stunning, the following results are obtained: all animals show respiratory arrest within 100 seconds. After being dipped in a CO2 enriched atmosphere for a few seconds the turkeys show head shaking and forced respiration which has to be considered as a strain the animals are conscious of. The meat hygienic results after CO2 stunning are much better than after electrical stunning. PMID- 8720799 TI - [Animal welfare and profit motive in compatible pet shops]. AB - In recent years, the zoo specialist shops have realized that their survival is depending from an adequate translation of animal welfare aspects, corresponding to its significance in our society. In particular, this includes that the shop's pet animals are presented adequately, that the offer of pet equipment is in conformity with animal welfare rules and that a competent consultation of the clients, including all animal species offered is ensured. PMID- 8720800 TI - [Species specific animal housing in pet shops (catalogue of measures)]. AB - Many veterinarians lack the essential zoological and ethologic knowledge of toy fishes and birds, reptiles, and the amphibians to correctly judge the specific keeping of animals in pet shops. To make up such deficits, the study group "AK 8 Zoofachhandel" (= study group 8 pet shops) of "Tierarztliche Vereinigung fur Tierschutz TVT" (= Veterinarian Consortium for the Protection of Animals) has been setting up lists, which contain criteria that enables veterinarians to judge the conditions of animal keeping in pet shops equally. Any kind of defect determined in an inspection of a pet shop will consequentially lead to the specific measures for each individual case. These measures have to be made out by the veterinarian in accordance with those lists and his individual knowledge. The following slides are examplary for good as well as bad keeping conditions. PMID- 8720801 TI - Cationic amphiphiles with G-protein-stimulatory activity: studies on the role of the basic domain in the activation process. AB - 2-Substituted histamines, which are cationic-amphiphilic, activate pertussis toxin-sensitive G-proteins by a receptor-independent mechanism. Lipophilicity is an important determinant for this G-protein activation, but the influence of basicity remained unclear. We prepared four imidazole-containing compounds and nine alkylamines with different basicity and studied their effcts on high affinity GTP hydrolysis in HL-60 membranes. The substances contained a lipophilic domain (L) and polar (P) and/or cationic (C) domains. Compared to 2-substituted histamines (L-P-C), the corresponding L-C-P type compounds were less potent and effective which, however, were still more active than L-P-P type substances. Among alkylamines, 2-octadecylaminoethyl-N,N-diethylamine (11) was the most potent GTPase activator (pEC50 = 5.7; 100% stimulation above basal). Conversely, 3-(5-hydroxyundecylamino)propylamine (15) was the most efficient compound (pEC50 = 4.3; 160% stimulation above basal). Compared to compounds 11 and 15, which belong to the L-C-C type, the corresponding L-P-C type compounds were less active. Thus, among 2-substituted histamines and alkylamines, a terminal cationic domain is critical for potent and/or effective receptor-independent G-protein activation. An L-C-X structure (X = P or C) is more favorable for G-protein activation than a L-P-X structure. Potency and efficiency of receptor-independent G-protein activators are independently determined drug parameters. PMID- 8720803 TI - On the synthesis of pyridazomycin congeners. Part 2: L-alpha-Amino acids bearing a terminal azinium or diazinium system. AB - The synthesis of a variety of novel compounds structurally related to the antimicrobial natural product pyridazomycin via alkylation of appropriate azine and diazine derivatives is reported. Based on the results of preliminary antimicrobial tests the dependence of antimicrobial activity from several structural features of pyridazomycin is discussed. PMID- 8720802 TI - Structure-activity relationship studies of CNS agents. Part 24: New analogs of N tert.-butyl-3-[4-(2-methoxyphenyl)-1-piperazinyl]-2-phenylpropanamide (WAY 100135). AB - A series of new N-substituted derivatives of 3-[4-(2-methoxyphenyl)-1 piperazinyl]-2-phenylpropanamide, 6-10, were synthesized and their 5-HT1A, 5 HT2A, and alpha 1 receptor affinities were determined. All the compounds were highly potent 5-HT1A ligands with a moderate or low 5-HT2A and alpha 1 affinity. It was shown that the 5-HT2A affinity of 1 and 6-10 depended crucially on the volume of amide substituents. None of the investigated racemic mixtures 1 and 6-8 antagonized the 8-OH-DPAT-induced lower lip retraction in rats, whereas (+/-)-7 behaved like a weak agonist of 5-HT1A receptors in the model used. PMID- 8720804 TI - Structural modifications of the primary amino group of anticonvulsant aryl semicarbazones. AB - A number of aryl semicarbazones had been shown previously to possess significant anticonvulsant properties. The principal objective of the present investigation was to determine the importance of the primary amino group in this series of compounds by replacing it with other substituents. The results indicate that the amino group was not essential for anticonvulsant activity. However its replacement by an aryl ring generally abolished activity while a terminal phenylamino function was better tolerated. Thus both the size of the group and its hydrogen bonding capabilities appear to influence bioactivity. Alteration of the oxygen atom of the semicarbazones by isosteres did not enhance anticonvulsant properties. PMID- 8720805 TI - An in vitro perfusion model for the determination of absorption properties of drugs in isolated rat small intestine. AB - A two-compartment model was developed to perform in vitro permeability studies of drugs. Preparations of surviving small intestine are continuously perfused under permanent oxygenation. The apparatus especially allows the usage of variable volumes of donor and adaptable length of intestinal segments ranging from 3 to 20 cm. Both the mucosal and the serosal side of the apparatus are open for sample collection and additional instrumentation. Due to the construction the use of high-surface activity compounds is possible. Data may be derived as absorption rates (micrograms.cm-1.min-1) and concentration increases vs. time profiles. PMID- 8720806 TI - Kinetics of release and simulated absorption of methyl nicotinate from different ointment formulations: in vitro-in vivo correlations. AB - The influence of the vehicle on release and simulated absorption of methyl nicotinate (MN) was evaluated using in vitro systems in order to find a correlation with data previously obtained in vivo. Simulation of drug absorption was carried out using a porous polymer membrane soaked with lipophilic phases such as n-dodecanol and isopropyl myristate. Ointment composition influenced differently both release and absorption of MN independent of drug concentration. The degree of skin redness induced by MN was found to be significantly correlated to in vitro simulated drug absorption but not to in vitro drug release. PMID- 8720807 TI - Formulation and in vitro examination of furosemide containing suppositories and preliminary experiences of their clinical use. AB - Rectal suppositories containing furosemide (4-chloro-N-furfuryl-5 sulfamoylanthranilic acid) and furosemide sodium were formulated with various suppository bases. The in vitro drug release of Massa Estarinum 299 proved to be the best from the vehicle having various physical-chemical properties. The diuretic effect of the two suppositories was compared in a prospective, crossover clinical trial including 8 patients. Both preparations have induced an increase of urine flow, which was comparable to the diuretic effect of the tablet. PMID- 8720808 TI - [Nisin, a potential preservative for topical preparations]. AB - The lantibiotic compound nisin has a bacterial effect on gram positive bacteria and was investigated for the possible use as a preservative in topical formulations. For this reason, its minimal inhibitory concentration (MIC) was determined in different types of ointments. Beside its effect on gram positive bacteria it had also a bactericidal activity on gram negative bacteria in concentrations of 0.06% (W/W). Within the first month there was no detectable reduction of nisin activity in different dermatological preparations. We conclude from these results that the combination of nisin with fungicidal substances represents a possible alternative to well known preservatives. PMID- 8720809 TI - [Synthesis and antimicrobial action of 4-arylideneamino-3-(alpha,alpha- diphenyl alpha-hydroxmethyl)-1,4-dihydro-5H-1,2,4-triazino-5-thiones and 6-arene-3 (alpha,alpha-diphenyl-alpha-hydroxymethyl)-7H-s-triazolo(3,4- b) (1,2,4)thiadiazine]. PMID- 8720810 TI - Inhibitory activity of isorhamnetin glycosides from Calendula officinalis L. on the activity of lipoxygenase. PMID- 8720811 TI - [Treatment of extravasation of intravenously administered agents]. PMID- 8720812 TI - [Cholesterol synthesis inhibitors: lowering of cholesterol for everybody?]. PMID- 8720813 TI - [Psychological and psychiatric problems in chronic somatic patients; recommendations for future studies and implications for care]. PMID- 8720814 TI - [The treatment of extravasation of drugs]. PMID- 8720815 TI - [Patient discharge criteria following surgery in day care]. PMID- 8720816 TI - [The effect of the wording of information about side effects on patients' interpretation of reported side effects]. AB - OBJECTIVE: To determine whether changes in the side effects frequency information in a patient information leaflet affects the interpretation of the frequency, the recall of this information and the number of side effects reported. DESIGN: Randomized controlled trial. SETTING: A field experiment was carried out in which three versions of a package insert concerning diclofenac were randomized and tested in 33, 30 and 34 patients respectively. Version 1 contained the original side effects information. In version 2 the frequency descriptors were manipulated (lowered, e.g. 'sometimes' was replaced by 'seldom'). This was because a preliminary investigation had shown that patients make higher assessments of side effect frequencies than the leaflet writers want to convey. In version 3 an introduction was added with general information on side effects. RESULTS: The frequency assessments differed enormously between subjects. Especially the lower frequencies were virtually impossible to convey by means of words. The lower ranking frequency descriptors did not only result in lower frequency assessments, but also led to lower recall score for side effects mentioned in the text and to a smaller number of side effects actually reported by patients. The only effect of the introduction was to lower the frequency assessments for the side effects following the introductory paragraph. CONCLUSIONS: Side effects with high assessed frequencies were read with more attention. The advantages and disadvantages of numerical frequency statements need to be discussed. PMID- 8720817 TI - [Electron beam irradiation is effective in the treatment of skin carcinomas; a comparison with superficial roentgen therapy]. AB - OBJECTIVE: To compare recurrence rates and cosmetic results after electron beam therapy versus superficial X-ray therapy for basal cell and squamous cell carcinomas of the skin. SETTING: Department of Radiotherapy, University Hospital Leiden, the Netherlands. DESIGN: Retrospective. METHODS: A retrospective analysis was performed of 389 histologically confirmed basal cell and squamous cell carcinomas of the skin (1980-1990), treated with either superficial X-rays or electron beam therapy, with regard to local recurrence rates and cosmetics scored with an arbitrary three point scale. RESULTS: Of 389 patients 19 developed a local recurrence (4.9%). For small tumours (irradiated surfaces < or = 10 cm2) a local recurrence rate of 2.2% was observed, both after electron beam therapy and after superficial X-ray therapy. Large tumours (irradiated surface > 50 cm2, treated with electron beam) had a higher recurrence rate (13.8%) as had tumours which had been treated before. The cosmetic results with electron beam treatment were better, because smaller fractions were used compared with superficial X-ray treatment. CONCLUSION: In our hands electron beam treatment was not inferior to superficial X-ray treatment, cosmetically even better and also applicable in case of larger tumours. The inferior results of electron beam therapy reported in literature may be due to technical factors. PMID- 8720818 TI - [A child with hyperpyrexia syndrome and multiple organ failure caused by and an orthopedic corset]. AB - A 4-year-old boy who each night was placed in an orthopaedic corset because of congenital multiple arthrogryposis developed heat stroke with hyperpyrexia and multiorganic failure in a hot summer. Treatment consisted of administration of fluids, antibiotics and anticonvulsive agents; the child recovered completely. In children who are sick and have fever, the use of such corsets should temporarily be suspended. PMID- 8720819 TI - [Computer-assisted education in problem-solving in neurology; a randomized educational study]. AB - OBJECTIVE: To determine the effect of computer-based medical teaching (CBMT) as a supplementary method to teach clinical problem-solving during the clerkship in neurology. DESIGN: Randomized controlled blinded study. SETTING: Academic Medical Centre, Amsterdam, the Netherlands. METHOD: 103 Students were assigned at random to a group with access to CBMT and a control group. CBMT consisted of 20 computer simulated patients with neurological diseases, and was permanently available during five weeks to students in the CBMT group. The ability to recognize and solve neurological problems was assessed with two free-response tests, scored by two blinded observers. RESULTS: The CBMT students scored significantly better on the test related to the CBMT cases (mean score 7.5 on a zero to 10 point scale; control group 6.2; p < 0.001). There was no significant difference on the control test not related to the problems practised with CBMT. CONCLUSION: CBMT can be an effective method for teaching clinical problem-solving, when used as a supplementary teaching facility during a clinical clerkship. The increased ability to solve problems learned by CBMT had no demonstrable effect on the performance with other neurological problems. PMID- 8720820 TI - [TIA... an aspirin?]. PMID- 8720821 TI - [TIA... an aspirin?]. PMID- 8720822 TI - [TIA... an aspirin?]. PMID- 8720823 TI - [Severe 4-aminopyridine poisoning in a body-builder]. PMID- 8720824 TI - [Opium: an old story]. PMID- 8720826 TI - [European drug registration and the Internet]. PMID- 8720825 TI - [Monoclonal antibodies for diagnosis and therapy in oncological patients]. PMID- 8720827 TI - Motional properties of a pentasaccharide containing a 2,6-branched mannose residue as studied by 13C nuclear spin relaxation. AB - 13C relaxation data obtained at three different magnetic fields, 9.4, 11.8 and 14.1 T, and at two temperatures, 303 and 318 K, are reported for the pentasaccharide p-trifluoroacetamidophenyl 2,6-di-O-[beta-D-galactopyranosyl-(1- >4)-O-2-acetamido-2-deoxy-beta-D- glucopyranosyl] alpha-D-mannopyranoside. The pentasaccharide consists of two disaccharide units, attached at positions 2 and 6 to the central mannopyranoside residue. The relaxation data were interpreted with the Lipari-Szabo model-free approach. For the central mannose residue in the molecule a high order parameter (S2 = 0.91) was found and the relaxation data could be interpreted with the truncated form of the Lipari-Szabo model. The motional behavior of the two 2-acetamido-2-deoxy-glucopyranoside residues was found to differ. The one attached at the primary hydroxylic position displayed more extensive local motion (S2 = 0.75-0.77) than the one attached at the secondary hydroxylic position (S2 = 0.83-0.85). More extensive local motion for the two outer galactopyranoside residues was found (S2 = 0.56-0.59), but no significant difference in motional behavior between the two residues could be observed. Analysis of the relaxation data for the exocyclic carbons confirmed the results for the rings. For the mannose C6, the same motional parameters as obtained for the substituting 2-acetamido-2-deoxy-glucopyranoside residue were found. The two exocyclic carbons on the 2-acetamido-2-deoxy-glucopyranoside residues showed more extensive local motion, with lower order parameters (S2 = 0.59-0.66). PMID- 8720828 TI - Comparison of the structure of human recombinant short form stromelysin by multidimensional heteronuclear NMR and X-ray crystallography. AB - Stromelysin-1 is a matrix metalloprotease that has been implicated in a number of degenerative diseases. Here we present the refined NMR solution structure of the catalytic domain of stromelysin-1 complexed with a small inhibitor and compare it to the X-ray crystal structure of the same complex. The structures are similar in global fold and show an unusual bottomless S1' subsite. There are differences, however, in the least well defined regions, Phe83-Ile89, His224-Phe232 and Pro249 Pro250, reflecting the lack of NOE data and large B-factors. The region His224 Phe232 contains residues of the S1' subsite and, consequently, small differences are observed in this subsite. Hydrogen-bond data show that, in contrast to the crystal structure, the solution structure lacks a hydrogen bond between the amide of Tyr223 and the carbonyl of the P3' residue. Analysis of bound water shows two tightly bound water molecules both in the solution and the crystal structure; neither of these waters are in the inhibitor binding site. PMID- 8720829 TI - Characterization of an artificial dimer of ribonuclease H using 1H NMR spectroscopy. AB - The protein fusion technique was applied in the synthesis of an artificial dimer of ribonuclease H (305 residues). 1H NMR spectroscopy was used to analyze the structure of this dimer. Spectral profiles and pKa values of the histidine residues obtained using 1H NMR indicate that the dimer retains the secondary and tertiary structures of the intact monomer. Selective spin-lattice relaxation measurements suggest that the two monomeric units in the dimer are in tight contact. Furthermore, the 2D 1H NMR and paramagnetic relaxation filter results show that the two monomers bind together through interactions between the N- and C-terminal sites of the linked regions. PMID- 8720830 TI - 1H NMR investigation of the secondary structure, tertiary contacts and cluster environment of the four-iron ferredoxin from the hyperthermophilic archaeon Thermococcus litoralis. AB - The solution molecular structure of the four-iron ferredoxin (Fd) from the hyperthermophilic archaeon Thermococcus litoralis (Tl) has been investigated by 1H NMR spectroscopy. TOCSY and NOESY experiments in H2O, tailored to detect both weakly and strongly relaxed resonances, together with steady-state NOEs in both H2O and D2O, allowed the identification of 58 of the 59 residues, with one residue near the paramagnetic center undetected. It is shown that the contact shifted and strongly relaxed signals for all four cysteines ligated to the paramagnetic cluster can be assigned by standard backbone connectivities that do not require any assumptions about the tertiary structure. Secondary structural elements identified in Tl Fd are a three-stranded antiparallel beta-strand involving the termini of the protein, a double beta-strand (also antiparallel), two alpha-helices and four turns. The existence of a disulfide bridge between the nonligated cysteines is also proposed. Dipolar contacts observed in the NOESY maps and by steady-state NOEs between the ligated cysteines and the 'diamagnetic' protein matrix indicate that the overall folding pattern of Tl Fd is very similar to that of the 3Fe ferredoxin from the mesophilic bacterium Desulfovibrio gigas [Kissinger et al. (1991) J. Mol. Biol., 219, 693-723]. The influence of the paramagnetism of the cluster on the relaxation properties of the proton signals of nonligated residues near the cluster, as well as on the ligated cysteines, correlates well with the proximity to the cluster iron(s), as predicted from the crystal structures for homologous protons of other single-cluster ferredoxins. Finally, the potential role of the various identified structural factors in contributing to the hyperthermostability of this protein is discussed. PMID- 8720831 TI - Validation of the use of intermolecular NOE constraints for obtaining docked structures of protein-ligand complexes. AB - The use of intermolecular NOEs for docking a small ligand molecule into its target protein has been investigated with the aim of determining the effectiveness and methodology of this type of NOE docking calculation. A high resolution X-ray structure of a protein-ligand complex has been used to simulate loose distance constraints of varying degrees of quality, typical of those estimated from experimental NOE intensities. These simulated data were used to examine the effect of the number, distribution and representation of the experimental constraints on the precision and accuracy of the calculated structures. A standard simulated annealing protocol was used, as well as a more novel method based on rigid-body dynamics. The results showed some analogies with those from similar studies on complete protein NMR structure determinations, but it was found that more constraints per torsion angle are required to define docked structures of similar quality. The effectiveness of different NOE constraint averaging methods was explored and the benefits of using 'R-6 averaging' rather than 'centre averaging' with small sets of NOE constraints were shown. The starting protein structure used in docking calculations was obtained from previous X-ray or NMR structure studies on a related complex. The effects on the calculated conformations of introducing structural differences into the binding site of the initial protein structure were also considered. PMID- 8720832 TI - Assignment of aliphatic side-chain 1HN/15N resonances in perdeuterated proteins. AB - The perdeuteration of aliphatic sites in large proteins has been shown to greatly facilitate the process of sequential backbone and side-chain 13C assignments and has also been utilized in obtaining long-range NOE distance restraints for structure calculations. To obtain the maximum information from a 4D 15N/15N separated NOESY, as many main-chain and side-chain 1HN/15N resonances as possible must be assigned. Traditionally, only backbone amide 1HN/15N resonances are assigned by correlation experiments, whereas slowly exchanging side-chain amide, amino, and guanidino protons are assigned by NOEs to side-chain aliphatic protons. In a perdeuterated protein, however, there is a minimal number of such protons. We have therefore developed several gradient-enhanced and sensitivity enhanced pulse sequences, containing water-flipback pulses, to provide through bond correlations of the aliphatic side-chain 1HN/15N resonances to side-chain 13C resonances with high sensitivity: NH2-filtered 2D 1H-15N HSQC(H2N-HSQC), 3D H2N(CO)C gamma/beta and 3D H2N(COC gamma/beta)C beta/alpha for glutamine and asparagine side-chain amide groups; 2D refocused H(N epsilon/zeta)C delta/epsilon and H(N epsilon/zeta C delta/epsilon)C gamma/delta for arginine side-chain amino groups and non-refocused versions for lysine side-chain amino groups; and 2D refocused H(N epsilon)C zeta and nonrefocused H(N epsilon, eta)C zeta for arginine side-chain guanidino groups. These pulse sequences have been applied to perdeuterated 13C-/15N-labeled human carbonic anhydrase II (2H-HCA II). Because more than 95% of all side-chain 13C resonances in 2H-HCA II have already been assigned with the C(CC)(CO)NH experiment, the assignment of the side-chain 1HN/15N resonances has been straightforward using the pulse sequences mentioned above. The importance of assigning these side-chain HN protons has been demonstrated by recent studies in which the calculation of protein global folds was simulated using only 1HN-1HN NOE restraints. In these studies, the inclusion of NOE restraints to side-chain HN protons significantly improved the quality of the global fold that could be determined for a perdeuterated protein [R.A. Venters et al. (1995) J. Am. Chem. Soc., 117, 9592-9593]. PMID- 8720833 TI - Do NOE distances contain enough information to assess the relative populations of multi-conformer structures? AB - The feasibility of determining the relative populations of multi-conformer structures from NOE-derived distances alone is assessed. Without cross-validation of the NOE restraints, any population ratio can be refined to a similar quality of the fit. Complete cross-validation provides a less biased measure of fit and allows the estimation of the correct population ratio when used in conjunction with very tight distance restraints. With the qualitative distance restraints most commonly used in NMR structure determination, cross-validation is unsuccessful in providing the correct answer. Other experimental sources are therefore needed to determine relative populations of multi-conformer structures. PMID- 8720835 TI - Through-bond correlation of imino and aromatic resonances in 13C-, 15N-labeled RNA via heteronuclear TOCSY. AB - Novel HCCNH TOCSY NMR experiments are presented that provide unambiguous assignment of the exchangeable imino proton resonances by intranucleotide through bond connectivities to the (assigned) nonexchangeable purine H8 and pyrimidine H6 protons in uniformly 15N-, 13C-labeled RNA oligonucleotides. The HCCNH TOCSY experiments can be arranged as a two-dimensional experiment, correlating solely GH8/UH6 and GH1/UH3 proton resonances (HCCNH), 51 as three-dimensional experiments, in which additional chemical shift labeling either by GN1/UN3 (HCCNH) or by GC8/UC6 (HCCNH) chemical shifts is introduced. The utility of these experiments for the assignment of relatively large RNA oligonucleotides is demonstrated for two different RNA molecules. PMID- 8720834 TI - Separation of intramolecular NOE and exchange peaks in water exchange spectroscopy using spin-echo filters. AB - A technique for separating intramolecular NOE and solvent-proton exchange peaks in exchange spectroscopy is demonstrated. This method utilizes the large differences in relaxation and coupling properties of water and macromolecules to separate the two effects. The spin-echo filter consists of a water-frequency selective 90 degrees pulse followed by a spin-echo sequence. If the echo time is sufficiently long, protein resonances (e.g. C alpha H protons) excited by the selective pulse are removed due to their much shorter T2 values and J-coupling evolution. By combining the filter with exchange spectroscopy (EXSY) or water exchange (WEX) filter experiments, exchange peaks can be selectively observed. In this paper the filter is combined with a modified version of the WEX filter (WEX II filter) with 1D and 2D detection and applied to a zinc finger peptide and to staphylococcal nuclease, allowing estimation of the contribution of intramolecular NOEs to the exchange spectra. PMID- 8720836 TI - [Non-conventional infectious agents or PRIONS]. PMID- 8720837 TI - [Oxalosis]. PMID- 8720838 TI - [Muscular involvement in the course of AIDS. Anatomo-clinical study of 17 cases]. AB - We report 17 patients seropositive for the human immunodeficiency virus, with muscle tissue involvement in different stages of the disease. Some patients are treated with azidothymidine (AZT). Others have no opportunistic infections. In all cases, there are some muscular symptoms such as progressive symetric and proximal muscular weakness with myalgias, elevated serum muscle enzymes, abnormal electromyogramma and very often a peripheral neuropathy. The muscle biopsy reveals the following features: rarely a focal muscular opportunistic infection in advanced stage of the disease is observed; a polymyositis is quite often the first clinical manifestation of the disease; a myopathy with mitochondrial involvement is observed in some of the AZT treated patients; some cachectic, under nourrished, bedridden patients present a type II muscle fiber atrophy. We conclude that a muscle biopsy could help us in our therapeutic planning directing us to a corticotherapy in the polymyositis, mitochondriopathies and wasting syndrome. Interruption alone of AZT or associated with a treatment by carnitine could allow remission of the muscular pathology. PMID- 8720840 TI - [Acinar cell carcinoma of the pancreas revealed by Weber-Christian syndrome]. AB - Acinar cell carcinoma of the pancreas is a rare tumor of the exocrine pancreas that may be associated with a characteristic syndrome of disseminated fat necrosis with a bad prognosis. We report a case of this tumor occurring in a 63 year-old man who presented with polyarthralgia and panniculitis of the legs. The histological features and the differential diagnosis of acinar cell carcinoma are discussed. PMID- 8720839 TI - [Hyperthyroidism induced by amiodarone and hyperthyroidism induced by iodine. Histologic, immunohistochemical and ultrastructural aspects]. AB - Amiodarone is a widely-used anti-arythmic drug that induces an iodine overload and, in 1 to 23% of the patients, a thyrotoxicosis. In a few cases, the thyrotoxicosis may be refractory to conventional pharmacological therapy, thus leading to thyroidectomy. We report the thyroid gland pathology in 5 thyroidectomies that were performed for uncontrolled thyrotoxicosis, induced by amiodarone (4 cases) and by Colchimax, another iodine-rich drug (1 case). Two cases have been studied by electron microscopy. Pathologic findings were: [1] colloid transformation of the parenchyma, [2] areas of follicular disruption with numerous foamy macrophages in the colloid, [3] regenerative areas and [4] a moderate T lymphocytic infiltration. No lysosomal lamellar inclusion body was found by electron microscopy. These lesions may be rather specific of an iodine toxicity. The pathogeny is still misunderstood, and probably complex. It may essentially involve toxic mechanisms, as well as immunologic or allergic mechanisms. PMID- 8720841 TI - [Adenocarcinoma in an ileal duplication]. AB - We describe a 54-year-old man with a history of multiple congenital abdominal malformations who developed an adenocarcinoma in a small intestinal duplication. To our knowledge this is the second reported case of an exceedingly rare complication of small intestinal duplication. The relevant literature is reviewed. PMID- 8720842 TI - [Laryngeal cytomegalovirus infection after heart transplantation]. AB - The authors report a case of laryngeal ulceration due to cytomegalovirus occurring in a 60-year-old man one month after heart transplant. The recipient was initially CMV seronegative and the donor positive. No preventive treatment was administered. One month after the transplantation, search for CMV in blood and urine became positive and a seroconversion occurred. Laryngeal biopsy showed an inflammatory reaction with cells containing atypical inclusions and positivities with anti-CMV antibody. Usual intranuclear eosinophilic inclusions of classic CMV infection were absent. The authors stress the unusual site of the lesions in this primo-infection and the atypical aspect of the inclusions. Such atypical inclusions which have already been reported in immunosuppressed people make the diagnosis more difficult. PMID- 8720843 TI - [Fortuitous discovery of an inguinal hernia]. PMID- 8720844 TI - [Spontaneous expulsion of a uterine mass]. PMID- 8720845 TI - [Thrombopenia resistant to corticosteroids revealing a non-hematologic disorder]. PMID- 8720846 TI - [Unilateral ovarian metastatic localization revealing a squamous cell carcinoma of the uterine cervix]. PMID- 8720847 TI - [Malignant tumors of the colon with two components (carcinosarcoma). Report of a case]. PMID- 8720848 TI - [In situ gene amplification on tissue sections (in situ PCR). A new technique for pathologists]. AB - In situ polymerase chain reaction is a recent technique which combines the sensitivity of PCR reaction to intracellular localization of genomic sequences with the same specificity as in situ hybridization. This reaction is based on the in situ annealing and polymerisation of oligonucleotides complementary to nucleotides located at each side of the target DNA sequence to amplify. We describe the Hot Start PCR (DNA) and the Hot Start PCR after reverse transcription step (RNA). It allows to amplify some nucleic sequences to a high level, becoming easier to detect. The vizualisation can be realized by direct in situ PCR, the product obtained being directly identifiable by incorporation of labeled nucleotides or primers, or preferentially by indirect in situ PCR. In this case, the amplification is followed by in situ hybridization with labeled probes. This last procedure is more specific. Numerous controls are essential at each step of the technique for validating results. PMID- 8720849 TI - Performance of a Graesser contactor in the continuous extraction of whey proteins: mixing, mass transfer and efficiency. AB - The performance of a Graesser Raining Bucket Contactor in an extraction using aqueous two-phase systems was characterized by axial mixing coefficients and counter current mass transfer for whey protein purification. The influence of rotor speed, phase ratio and phase velocity on the salt phase axial mixing coefficients was determined applying a dispersion model to the residence time distribution experiments. When dissolving whey powder in the salt-rich phase of the system, alpha-lactalbumin was extracted to the polyethylene-glycol-rich phase, whereas beta-lactoglobulin predominantly remained in the bottom phase, making the process suitable for removal of the major allergen beta-lactoglobulin. The mass transfer in the phase system was determined from the steady state data of alpha-lactalbumin extraction. On the basis of a diffusion model, the efficiency of the counter current extraction under different process conditions was calculated from results of the mass transfer experiments. PMID- 8720850 TI - Using differential scanning calorimetry to elucidate metal-protein binding sites in alpha- and gamma-chymotrypsin. AB - Crystalline alpha-chymotrypsin preparations are contaminated by the post translational variant, gamma-chymotrypsin. The contaminant can account for 5-50 weight percent of the preparation based on thermal analysis. Such contamination can be problematic because this serine protease has both commercial and deactivation model system utility, and the presence of the contaminant may not be detectable by activity assays. Prior work has shown that simple pH gradient elution can separate the two chymotrypsins when loaded to a Cu(2+)-IMAC column; gamma-chymotrypsin eluted first indicating that its interaction with immobilized Cu2+ is weaker. The molecular features that endow these serine proteases with metal affinity has been investigated further by performing differential scanning calorimetry (DSC) studies in the presence and absence of Cu2+, and at different pH values. The dependence of thermostability on pH for fixed metal concentration reveals an interplay between stabilizing and destabilizing metal binding events. The results are consistent with Cu(2+)-chymotrypsin interaction occurring, in part, through binding to a glutamate- or aspartate-containing chelation site. The strength of this site may differ in the two chymotrypsins. PMID- 8720851 TI - Prediction of protein displacement by simplified immobilized metal ion affinity chromatographic model. AB - A simplified theoretical model was developed to predict complex behavior in protein displacement systems of immobilized metal ion affinity chromatography (IMAC). In this paper, experimental techniques for estimating equilibrium parameters were developed. The equilibrium parameters obtained were used to simulate displacement processes and to investigate the effect of mobile phase modifier (MPM) gradients in the IMAC separating systems. In the induced MPM gradient conditions, the solute movement analysis was able to predict the displacement separation well characterized by the intersections of the operation line under the effective displacer concentrations. The theoretical and experimental results demonstrate the utility of the IMAC model in predicting induced mobile phase modifier gradients and complex behavior in metal affinity displacement chromatography. Furthermore, the development of a rapid method for obtaining displacement profiles facilitates methods development and optimization of IMAC displacement separations. PMID- 8720852 TI - Neurosteroidal modulation of social isolation-induced decrease in pentobarbital sleep in mice. AB - Stressful manipulations are known to change the level of neurosteroids capable of interacting with GABAA receptor in the brain. To clarify the involvement of these neurosteroids in social isolation stress-induced decrease in pentobarbital sleep in mice, we examined the effects of 3 beta-hydroxy-5-pregnen-20-one-3-sulfate (pregnenolone sulfate, PS), a steroidal GABAA antagonist, and 3 alpha,21 dihydroxy-5 alpha-pregnan-20-one (allo-THDOC) and 3 alpha,21-dihydroxy-5 beta pregnan-20-one (THDOC), positive allosteric modulators of the GABAA receptor, on the hypnotic activity of pentobarbital in socially isolated mice. Pentobarbital (50 mg/kg, i.p.)-induced sleep was significantly shorter in isolated mice than in group-housed mice and adrenalectomy had no effect on the decrease of pentobarbital sleep following social isolation. PS (5-10 mg/kg, i.p. or 12-24 nmol, i.c.v.) decreased pentobarbital sleep in group-housed mice in a dose dependent manner without affecting the sleep in socially isolated mice. In contrast, allo-THDOC (14.9-37.4 nmol, i.c.v.) and THDOC (5-12.5 mg/kg, i.p. or 14.9-37.4 nmol, i.c.v.) reversed the pentobarbital sleep decreased by social isolation to the level in group-housed mice. These steroids had no effect on the pentobarbital sleep in group-housed mice. Such a reversing effect of THDOC in isolated mice was significantly blocked by PS (24 nmol, i.c.v.). Moreover, i.c.v. injection of yohimbine (30 nmol), methoxamine (200 nmol) and CRF (2.1 nmol) significantly decreased pentobarbital sleep in group-housed mice but not that in isolated mice. The effects of these drugs on pentobarbital sleep in group-housed mice were significantly attenuated by THDOC (12.5 mg/kg, i.p.). These results suggest that changes in the level of neurosteroids with ability to modulate GABAA receptor function are involved in social isolation-induced decrease in the hypnotic activity of pentobarbital in mice. PMID- 8720853 TI - Developmental and activity-dependent changes in K+ currents in satellite glial cells in mouse superior cervical ganglion. AB - Voltage-gated K+ currents were recorded from freshly dissociated satellite glial cells wrapping around ganglion cells in mouse superior cervical ganglion (SCG) by whole-cell recordings of patch clamp techniques. Both inward and outward K+ currents during membrane hyperpolarization and depolarization were observed in these glial cells. The current-voltage relation of these K+ currents became almost linear in cells obtained more than 4 weeks after birth. The magnitude of the density of inward K+ currents, which were elicited during membrane hyperpolarization and were eliminated by external barium, progressively increased during the first month after birth. This developmental increase in the magnitude of inward K+ current density was not affected by decentralization of SCG done by transection of cervical sympathetic trunk (CST) 5 days after birth. In adult mice, the magnitude of the inward K+ current density decreased after chronic conduction blockade of CST by local application of tetrodotoxin. On the other hand, the magnitude of the inward K+ current density increased after daily intraperitoneal injection of reserpine and this increase was abolished by pre treatment of decentralization of SCG. These results suggested that preganglionic innervation was not prerequisite for developmental increase in the inward K+ currents and preganglionic neuronal activity upregulates the inward K+ currents in adult mice. Neuronal regulation of glial K+ channel expression would assist in K+ clearance from periganglionic space to maintain neuronal activity. PMID- 8720854 TI - Influence of age on the clearance of K+ from the extracellular space of rat hippocampal slices. AB - We examined the hypotheses that aging alters the capacity of brain tissue to regulate extracellular K+ activity (K+o), and that age-related decreases in glucose metabolism may underlie these alterations. Hippocampal slices from young adult (6-9 months old), middle-aged (16-19 months old), and aged (26-29 months old) Fischer 344 rats were exposed to physiological solutions containing 5-20 mM glucose maintained at 36-37 degrees C. Schaffer collaterals in each slice were stimulated at 40 Hz for 2 s, and the resulting changes in K+o were recorded with K+o-sensitive microelectrodes placed in stratum pyramidale of hippocampal subfield CA1. We found that K+ clearance from the extracellular space of hippocampal slices was significantly slowed in the middle-aged group compared with the young adult group in physiological solutions containing 5 and 10 mM glucose. Age-related differences in K+o clearance disappeared in 20 mM glucose. Also, the rate of K+o clearance was modified by glucose concentration. These results suggest that K+ transport rates are modified by age, and that age-related alterations in glucose metabolism may be involved. PMID- 8720855 TI - Administration of amphetamine does not increase the functional efficacy of dopaminergic grafts made in infancy. AB - Previous reports have evoked the possibility that a priming stimulation of grafted dopaminergic (DA) neurones by amphetamine enhances their efficacy in behavioural tests performed several days later. The present study was designed to test this hypothesis. Five days after the unilateral destruction of the DA mesotelencephalic system of 3-day-old rat pups, DA grafts were implanted into the denervated neostriatum of half of the lesioned pups. At adulthood, lesion and graft groups were subdivided into 4 subgroups which received one of the following treatments: saline or amphetamine injection in an environment where the behavioural test was subsequently conducted (paired environment) or in an unrelated environment (unpaired environment). Five days later, rotational response to a tail-pinch stress was tested in the paired environment. In these conditions, we found no evidence for a priming effect of amphetamine. Animals that received amphetamine or saline in the unpaired environment displayed the same rotational response to the tail-pinch stress. On the other hand, a conditioning influence of the environment was detected. Thus, the effect previously described might have been caused by a conditioning effect and/or might be due to differences in the experimental conditions. This suggests that 'priming' the graft with amphetamine does not provide a general strategy to enhance the functional efficacy of DA grafts. PMID- 8720856 TI - Interactions of tetramethrin, fenvalerate and DDT at the sodium channel in rat dorsal root ganglion neurons. AB - Type I and type II pyrethroids and dichlorodiphenyltrichloroethane (DDT) are known to modulate the sodium channel to cause the hyperexcitatory symptoms of poisoning in animals. However, since the degrees to which neuronal sodium channel parameters are altered differ, a question is raised as to whether these insecticides bind to the same site in the sodium channel. Competition patch-clamp experiments were performed using rat dorsal root ganglion neurons which are endowed with tetrodotoxin-sensitive and tetrodotoxin-resistant sodium channels. D trans-Tetramethrin, S,S-fenvalerate and p,p'-DDT caused a slowly rising and slowly falling tail current to be developed in tetrodotoxin-sensitive sodium channels. In tetrodotoxin-resistant sodium channels, these insecticides, particularly tetramethrin and fenvalerate, generated a large and prolonged tail current upon repolarization. The effects of tetramethrin were reversible after washing with drug-free solution, whereas the effects of fenvalerate and DDT were irreversible. When fenvalerate application was followed by tetramethrin application, the characteristic changes in current by fenvalerate disappeared and the characteristic changes by tetramethrin appeared. After washout, the characteristic current pattern of fenvalerate reappeared. These results can be explained by assuming that the tetramethrin molecule displaces the fenvalerate molecule from the same binding site in the sodium channel protein, or that tetramethrin and fenvalerate bind to separate sodium channel sites which interact allosterically with each other. DDT interacted with fenvalerate and tetramethrin in the same manner. PMID- 8720857 TI - Differential regulation, by MK-801, of dopamine receptor gene expression in rat nigrostriatal and mesocorticolimbic systems. AB - Glutamate agonists have been shown to stimulate striatal dopamine release, but less is known about dopamine-glutamate interactions at the receptor level. We treated rats with 0.3, 1.0, or 3.0 mg/kg of MK-801, an NMDA antagonist, daily for 1 week and, using in situ hybridization, measured dopamine receptor mRNA levels in cortical and subcortical structures. MK-801 caused a significant increase of D1 and D2 mRNA in the dorsal and ventral striatum, a significant decrease of D3 mRNA in the nucleus accumbens, and a significant decrease of D1 mRNA in the limbic cortex. Dopamine autoreceptor expression, reflected by D2 mRNA in the midbrain, was increased in the ventral tegmental area, but not in the substantia nigra. Thus, MK-801 appears to differentially regulate the mesocorticolimbic and nigrostriatal dopamine systems. PMID- 8720858 TI - Dissociation of Fos-like immunoreactivity in lamina terminalis and magnocellular hypothalamic nuclei induced by hypernatremia. AB - Rats were given either slow (1 h) or rapid (10 min) intravenous infusions of either 6 or 12 mmol NaCl/kg body weight. Fos-like immunoreactivity (FLI) induced by the infusions was measured in several brain regions. The higher dose of NaCl induced FLI in structures of the lamina terminalis, including organum vasculosum (OVLT) and subfornical organ (SFO), as well as in the magnocellular supraoptic (SON) and paraventricular hypothalamic (PVN) nuclei. The lower dose of hypertonic NaCl induced FLI in only the SON and PVN. Faster delivery of the solute load tended to amplify the FLI in SFO and OVLT. These data confirm and extend previous reports of osmotically-induced FLI in rat brain and demonstrate that the discrepancies between these studies result from different dosage regimens of NaCl. The data are discussed as they relate to the lamina terminalis as a primary osmosensitive region in brain. PMID- 8720859 TI - Chronic reduction of cerebral blood flow in the adult rat: late-emerging CA1 cell loss and memory dysfunction. AB - Ten-month-old rats were subjected to permanent bilateral occlusion of both common carotid arteries (2-VO) to chronically but moderately reduce brain blood flow. 2 VO impaired Morris water maze acquisition as soon as 7 days post-surgery. 2-VO also caused a later-appearing impairment on the radial arm maze which did not reach significance until 63 days post-surgery. At 14 dats post-surgery there were no effects of 2-VO on hippocampal CA1 pyramidal cell number or density of glial fibrillary acidic protein (GFAP). Hippocampal choline acetyltransferase activity at 70 days was also unaffected by 2-VO. At 190 days post-surgery, however, the 2 VO rats showed loss of cells and increased GFAP density in CA1. The increased density of hippocampal GFAP correlated with radial arm maze but not Morris water maze impairment. It is suggested that 2-VO causes neuronal dysfunction which can be exacerbated by stress and thereby manifested on aversively motivated tasks such as the water maze. As well, CA1 neurons begin to degenerate after several weeks of the reduced energy availability caused by 2-VO and this impairs memory. Since reduced neuronal energy metabolism is associated with the progressive neurodegeneration that underlies disorders such as Alzheimer's, research should further explore the possibility that the effects of 2-VO may model age-related dementia. PMID- 8720860 TI - Differential abundance of superoxide dismutase in interneurons versus projection neurons and in matrix versus striosome neurons in monkey striatum. AB - To investigate whether differences in vulnerability to free radicals might underlie differences among striatal neurons in their vulnerability to neurodegenerative processes such as occur in ischemia and Huntington's disease, we have analyzed the localization of superoxide free radical scavengers in different striatal neuron types in normal rhesus monkey. Single- and double-label immunohistochemical experiments were carried out using antibodies against the enzymes copper, zinc superoxide dismutase (SOD1), or manganese superoxide dismutase (SOD2), and against markers of various striatal cell types. Our results indicate that the striatal cholinergic and parvalbumin interneurons are enriched in SOD1 and/or SOD2, whereas striatal projection neurons and neuropeptide Y/somatostatin (NPY+/SS+) interneurons express only low levels of both SOD1 and SOD2. We also found that projection neurons of the matrix compartment express significantly higher levels of SOD than those in the striosome compartment. Since projection neurons have been reported to be more vulnerable than interneurons and striosome neurons more vulnerable than matrix neurons to neurodegenerative processes, our results are consistent with the notion that superoxide free radicals are at least partly involved in producing the differential neuron loss observed in the striatum following global brain ischemia or in Huntington's disease. PMID- 8720861 TI - Sex differences in corticosteroid binding in the rat brain: an in vitro autoradiographic study. AB - Several previous studies have raised the possibility of sex differences in the distribution of corticosteroid receptors in the brain. The direction and magnitude of these differences have, however, remained controversial. In the present study, we have re-examined the concentrations of mineralocorticoid (MR) and glucocorticoid (GR) receptors in the brains of male and female rats at varying times (1 to 6 days) after combined gonadectomy (GDX) and adrenalectomy (ADX). Cytosol binding assays confirmed the presence of higher MR levels in short term (3-day) GDX-ADX males. This difference disappeared by 6 days after surgery, as receptor levels in females rose to be equivalent to those in males. Using an improved in vitro autoradiographic method, the distribution of MR and GR was studied in males and females 3 days after GDX-ADX. The distribution of MR and GR in the brains of these rats was similar in the two sexes. MR binding in the male, however, was significantly greater than that in the female throughout the principal cell fields of the hippocampus. Measurements of circulating corticosterone levels at the time of GDX-ADX suggest that this sex difference may reflect a more rapid recovery of the MR system in males than in females following the stress-induced rise in corticosterone secretion occurring at the time of surgery. PMID- 8720863 TI - Quantitative analysis of contralateral hemisphere hypertrophy and sensorimotor performance in adult rats following unilateral neonatal ischemic-hypoxic brain injury. AB - The immature nervous system is capable of considerable compensatory reorganization following injury. This has been studied extensively following many different types of injury in humans and laboratory animals. One common risk factor associated with perinatal brain injury that has been associated with such reorganization is an ischemic-hypoxic event. Using the established Levine model of neonatal ischemic-hypoxia (IH) to create unilateral striatal, cortical and hippocampal damage, we investigated anatomical changes in the undamaged hemisphere contralateral to the injury. Specifically, we measured cross-sectional area (mm2) of brain sections at the level of +1.20 and -2.12 mm from bregma. In addition, we examined sensorimotor deficits in these animals during development and as adults by measuring the amount of time that the animals were able to remain on a rotating treadmill. Our results show that some animals exhibited hypertrophy in the hemisphere contralateral to the lesion as compared to measurements taken from normal control animals. Additionally, we have demonstrated that, following IH, animals that showed significant contralateral whole-hemisphere hypertrophy were able to remain on the Rota-Rod treadmill significantly longer than the animals that did not exhibit this hypertrophy. We conclude that there are compensatory reorganizational changes that occur in the undamaged hemisphere contralateral to injury in some animals following neonatal ischemic-hypoxic brain injury. Furthermore, our data suggest that this plasticity in the contralateral hemisphere may be functionally advantageous. PMID- 8720862 TI - Hypoxic neuronal damage in the absence of hypoxic depolarization in rat hippocampal slices: the role of glutamate receptors. AB - The propensity of neurons to undergo profound and precipitous depolarization is believed to contribute to their characteristic vulnerability to hypoxic injury. The length of time a neuron spends in a depolarized state following hypoxic depolarization (HD) is a critical determinant of the extent of irreversible cell damage. It is less clear, however, what the effects of moderate hypoxia are when HD does not occur. The present study examined the effects of prolonged, moderate hypoxia which does not elicit HD in rat hippocampal slices. Extracellularly recorded population excitatory postsynaptic potentials (pEPSPs) in stratum radiatum of CA1 were eliminated 10-15 min after initiating hypoxia. Physiological damage was related to the hypoxic duration: full, intermediate, or poor recovery of pEPSP slope was observed after 30, 60, or 120 min of hypoxia, respectively. The glutamate receptor antagonists, D,L-2-amino-5-phosphonovaleric acid (APV) or 6,7-dinitroquinoxaline-2,3-dione (DNQX), enhanced the post-hypoxic recovery of synaptic responses. These findings demonstrate that profound HD is not necessary to elicit physiological damage during moderate hypoxia; moreover, the neuroprotective actions of excitatory transmitter antagonists are not limited to their capacity to delay HD. The precise characterization of cellular responses under these conditions will be of particular importance for understanding the pathophysiology of an ischemic penumbra. PMID- 8720864 TI - Electrophysiological correlates of prior training: an in vitro study of an area of the avian brain which is essential for early learning. AB - Day-old domestic chicks will peck at any small, distinct object, such as a metal bead. One-trial passive avoidance learning can be established by coating the metal bead with methyl anthranilate (MeA) and allowing the birds to peck it once, after which they conspicuously avoid it. We have used birds successfully trained not to peck metal beads, and a control set of chicks where the training beads were innocuously dipped in water. Brain slices were prepared from both groups, containing the left, intermediate, medial part of the hyperstriatum ventrale (IMHV)--a region essential for this form of early learning. The electrophysiological properties of neurones in the IMHV were examined in vitro. Neurones recorded intracellularly in slices taken from MeA-trained birds had higher membrane resistances than did cells from water-trained controls. MeA training was also associated with an increased incidence of spontaneous, large EPSPs. Field responses to local electrical stimulation appeared to be somewhat greater in MeA-trained birds than in water-trained controls. In contrast, field potentials proved harder to potentiate with a burst of relatively high frequency stimulation in MeA-trained birds: the change in amplitude was less in MeA-trained birds, and there was less variability than in slices from water-trained controls. PMID- 8720865 TI - Cholinergic denervation-like changes in rat hippocampus following developmental lead exposure. AB - We investigated the effects of developmental lead exposure from embryonic day 16 (E16) through postnatal day 28 (PN28), on cholinergic and catecholaminergic markers in the septohippocampal pathway in rats through fourth month of age. Lead exposure resulted in a persistent 30-40% reduction of [3H]hemicholinium-3 ([3H]HC 3) binding in the hippocampus through PN120, and 20-30% reduction of septal and hippocampal choline acetyltransferase (ChAT) activity which persisted through PN84 but returned to control levels in both septum and hippocampus at PN112. The muscarinic ligand [3H]quinuclidinyl benzylate ([3H]QNB) binding was reduced in the septum at PN28 but did not differ significantly from controls at PN56-PN112. Neither short- nor long-term effects of Pb exposure on [3H]QNB binding were seen in the hippocampus. Similar to the effects of fimbria-fornix transection, Pb exposure resulted in a long-term 50-90% increase of tyrosine hydroxylase(TH) activity in the hippocampus, although neither treatment affected TH activity in the septum. The lead-induced increase in hippocampal TH was significantly attenuated by superior cervical ganglionectomy. It is concluded that the effects of perinatal lead exposure resemble in several respects those seen following surgical disruption of the septohippocampal pathway in adult animals. The denervation-like effects in the hippocampus may be an important factor in long term learning and cognitive impairments following developmental exposure to low levels of lead. PMID- 8720866 TI - Medial septal unit firing characteristics following injections of 8-OH-DPAT into the median raphe nucleus. AB - Extracellular single-unit recording techniques were used to examine the firing characteristics of neurons in the medial septum/diagnol band of Broca complex (MS/DB) following injections of the 5-HT1A agonist, 8-OH-DPAT, into the median raphe nucleus (MRN) of urethane-anesthetized rats. It had previously been shown that MRN injections of 8-OH-DPAT produce hippocampal theta rhythm. Injections of 8-OH-DPAT into the MRN produced a change in firing characteristics of MS/DB neurons from an irregular discharge to a pattern of rhythmical bursting in synchrony with hippocampal theta rhythm. Cross-correlational and coherence analyses demonstrated that the rhythmical firing pattern of MS/DB neurons strongly correlated with rhythmical fluctuations in the hippocampal EEG during periods of hippocampal theta produced by 8-OH-DPAT injections, but not during baseline conditions (i.e. hippocampal desynchronization). The results suggest that MRN control of the hippocampal EEG is modulated by the MS/DB. Serotonergic projections from the MRN to the MS/DB may normally act to inhibit the rhythmical bursting of MS/DB neurons, thereby producing hippocampal desynchronization. Suppression of MRN 5-HT neurons by MRN injections of 8-OH-DPAT would disinhibit MS/DB neurons, allowing them to burst rhythmically and thereby produce hippocampal theta rhythm. PMID- 8720867 TI - PKC-dependent reduction of the acetylcholine-evoked inward Na current in Aplysia D-neurons: effect of injected PKC and PKC activators. AB - The effect of elevated PKC activity on the membrane depolarization (D-response) evoked by extracellular ACh, applied on the soma of Aplysia neurons, was studied. Intracellularly injected PKC and certain PKC activators were used to elevate PKC activity. ACh-induced current was measured in voltage clamp. The neurons were treated extracellularly with the PKC activators: PDAc, SC-10, R-59949, (-)-ILV; or with purified PKC injected into the neuron through the recording electrode. PKC injection and treatment with any of the PKC activators caused a similar reduction of the ACh-induced inward Na current response (corresponding to D response), while the non-activating alpha-PDD had no effect. The results provide evidence that a PKC-dependent reduction of receptor responses also exists in this kind of Aplysia neurons. Furthermore, they show that the reduction of ACh response is indeed due to PKC activation (and not to a direct action of the phorbol ester). PMID- 8720868 TI - Brain-mediated responses to vaginocervical stimulation in spinal cord-transected rats: role of the vagus nerves. AB - The present study was designed to ascertain whether the vagus nerves convey functional sensory activity from the reproductive tract in rats. Previously, vaginocervical mechanostimulation (VS) was shown to increase pupil diameter (PD) and the threshold of vocalization to tail shock (Voc-T). These responses were attenuated but not abolished by combined bilateral transection of the 'genito spinal' nerves (i.e. pelvic, hypogastric and pudendal). Subsequent bilateral vagotomy further reduced or abolished the residual responses. In the present study, spinal cord transection above the known level of entry of the genito spinal nerves was combined with bilateral vagotomy. In ovariectomized rats, after spinal cord transection at thoracic 7 (T7X), lumbar 5 (L5X) levels, or sham surgery (Sh), responses to VS were measured, the vagus nerves were then transected bilaterally, and responses to VS were again measured. VS significantly increased Voc-T and PD after sham procedure or spinal cord transection at either level. Subsequent bilateral vagotomy abolished the VS-induced increase in PD in the T7X group. Due to low survival rate, the effect of vagotomy on Voc-T could not be determined. Consequently, we performed a second experiment. In non ovariectomized rats, VS significantly increased PD but reduced Voc-T in the T7X group compared to the Sh group, and subsequent bilateral vagotomy abolished both responses. These findings provide evidence that, in the rat, the vagus nerves provide a functional sensory pathway from the reproductive tract directly to the medulla oblongata of the brain, bypassing the spinal cord. PMID- 8720869 TI - Substance P receptor regulates the photic induction of Fos-like protein in the suprachiasmatic nucleus of Syrian hamsters. AB - Substance P (SP) is a candidate neurotransmitter or neuromodulator for conveying light information from the retina to the hypothalamic suprachiasmatic nucleus (SCN) where a circadian oscillator(s) is located in mammals. Immediate early gene c-fos has been demonstrated to be induced in the SCN with a brief light exposure at the subjective night, and suggested to play an important role in the photic entrainment of the oscillator. To clarify the possibility of an involvement of the SP receptor in the photic-induction of c-fos in the SCN, we examined effects of a SP receptor antagonist, spantide, on the light-induced Fos-like protein immunoreactivity (Fos-lir) in the SCN of Syrian hamster. The light-induced Fos lir was inhibited with the pretreatment of spantide in a dose-related manner and in an anatomically distinctive way. The higher dose of spantide (8 nmol) blocked light-induced Fos-lir substantially in the rostral and central areas of the SCN, and in the dorsal portion of the caudal SCN. However, it blocked Fos-lir only slightly in the ventral portion of the caudal SCN. These results suggest that the SP is involved in conveying light information to induce Fos protein in the hamster SCN, and that different neurotransmitter systems are involved in the light-induced Fos-lir in the different portions of hamster SCN. PMID- 8720870 TI - Age-related loss of cholinergic-muscarinic coupling to PLC: comparison with changes in brain regional PLC subtypes mRNA distribution. AB - Activation of phospholipase C (PLC) coupled to phosphoinositide (PtdIns) hydrolysis occurs through one of the two pathways. One of the major pathways for the neurotransmitter signaling involves phosphoinositide (PtdIns) specific and G protein dependent PLC-beta, which stimulates the formation of inositol triphosphate (IP3) and inositol tetraphosphate (IP4). Another pathway through the stimulation of calcium influx can directly activate all of the PLC isozymes. At least three isozymes of PLC have been characterized in the brain; PLC-A (alpha), PLC-I (beta) and PLC-II (gamma), which are shown to be localized differentially in brain regions. Muscarinic-cholinergic signals are mediated in large part through the hydrolysis of PtdIns by PLC. To investigate changes in muscarinic coupling to PLC during aging, we examined carbachol stimulated and calcium stimulated PtdIns hydrolysis in cerebral cortical membranes in young, middle aged and old rats. In order to determine whether PtdIns hydrolysis changes correspond to PLC isozyme expression in these animals, we examined three subtypes of PLC mRNA expression in brain sections of young and old rats using in situ hybridization technique. Our study indicated decreased carbachol-induced PLC activity in the cerebral cortex and, in contrast, increased PLC-beta mRNA in the frontal cortex and superficial cortical layer of aged rats. PLC-alpha mRNA was decreased in hippocampal regions of older rats. These studies suggest that during aging there is an uncoupling of muscarinic stimulated PtdIns hydrolysis, which is accompanied by an increased PLC-beta mRNA and decreased PLC-alpha mRNA that may represent compensatory changes in PLC expression. PMID- 8720871 TI - Amiloride inhibition of chorda tympani responses to NaCl and its temperature dependency in mice. AB - Inhibitory effects of amiloride on salt responses of the chorda tympani nerve and its temperature dependency were compared among three inbred strains of mice (C57BL, BALB and 129). In C57BL mice, lingual treatment with amiloride significantly suppressed responses to 0.1-1.0 M NaCl at two different temperatures, 24 +/- 2 degrees C and 12 +/- 2 degrees C. The magnitude of the amiloride-inhibited component of NaCl response was slightly larger at the higher temperature. In contrast, in BALB mice, amiloride suppression of NaCl responses was observed only at the lower temperature. No such suppression was exhibited by 129 mice at either temperature levels. These results suggest that there exist at least two different amiloride-sensitive receptor components for NaCl in mice: one is more sensitive to NaCl at the higher temperature, and the other is more sensitive at the lower temperature. It is hypothesized, C57BL mice possess the former (or both) component(s), whereas BALB mice have the latter one. The 129 strain may be lacking both components. PMID- 8720872 TI - Dual effects of thrombin and a 14-amino acid peptide agonist of the thrombin receptor on septal cholinergic neurons. AB - We have compared the effects of thrombin and of the 14-amino acid peptide agonist (TRAP-14) of the thrombin protease activated receptor (PAR) on cholinergic neurons in pure cultures of rat septal neurons and in co-cultures of septal neurons and glial cells. In pure septal cultures, low concentrations of thrombin (up to 10 nM) did not affect choline acetyltransferase (ChAT) activity, a marker of cholinergic neurons, or 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) reduction, an index of cell viability. However, 100 nM thrombin decreased ChAT activity and MTT reduction by 44 and 17%, respectively. In co cultures, a low concentration of thrombin (1 nM) increased ChAT activity (+75%), whereas a high concentration (100 nM) decreased it (-83%). At this high concentration, thrombin was neurotoxic, as indicated by a large decrease in MTT reduction (-80%). Thrombin effects on ChAT activity were mimicked by TRAP-14 both in pure septal cultures (no effect at 0.1 microM and -63% at 100 microM) and in co-cultures (+25% at 0.1 microM and -28% at 100 microM). In contrast, this peptide did not affect MTT reduction. These dual effects of thrombin and TRAP-14 on ChAT activity in co-cultures, were also observed on pure cultures of septal cells supplied with NGF. The activation and inhibition by TRAP-14 of the expression of ChAT activity in septal neuron/glial cell cultures were inhibited by a 9-amino acid peptide antagonist of thrombin PAR. Thus, the effects of thrombin on cholinergic neurons seem to be mainly mediated by thrombin PAR and glial cells seem to play a major role in these thrombin actions. PMID- 8720873 TI - Changes in NADPH diaphorase activity in forebrain structures of the laminae terminalis after chronic dehydration. AB - The effect of 3-day chronic dehydration on nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-diaphorase) histochemical staining in forebrain circumventricular structures was investigated in the rat. Increased number and/or intensity of staining of NADPH-diaphorase positive neurons was observed in subfornical organ, organum vasculosum of the laminae terminalis, and median preoptic nucleus. In addition, dense punctate NADPH-diaphorase reaction product was found throughout the internal portion of median eminence. These data suggest the involvement of nitric oxide synthase producing neurons in homeostatic mechanisms controlling body fluid balance and the circulation. PMID- 8720874 TI - Stimulation of 5-HT2A receptors in the paraventricular hypothalamus attenuates neuropeptide Y-induced hyperphagia through activation of corticotropin releasing factor. AB - The effect of 5-HT1 and 5-HT2 receptor agonists administered into the paraventricular hypothalamus was studied on the hyperphagia caused by neuropeptide Y (NPY) injected into the same area. The 5-HT2A/2C receptor agonist DOI (10-20 nmol/0.5 microliter) significantly reduced NPY overeating while the 5 HT1A/1B receptor agonist RU 24969 (3.5-14 nmol/0.5 microliter) and the 5-HT1B/2C receptor agonist mCPP (5-20 nmol/0.5 microliter) had no such effect. The 5-HT2A receptor antagonist spiperone (5 microgram/0.5 microliter) and the corticotropin releasing factor antagonist alpha-helical-CRF9-41 (0.5-1 micrograms/0.5 microliter) completely antagonized the effect of 10 nmol DOI. PMID- 8720875 TI - Circadian regulation of melatonin production in cultured zebrafish pineal and retina. AB - Melatonin release was measured from zebrafish pineal organs and retinas maintained in flow-through culture. Pineal organs released melatonin in a strong circadian rhythm through 5 days in constant darkness, and the phase of this rhythm was reset by in vitro exposure to phase-shifted light cycles. In contrast, the retinal melatonin rhythm rapidly damped out in constant darkness, even in the presence of (phase-shifted) light cycles. The zebrafish pineal should be useful for in vitro studies of vertebrate circadian clock mechanisms. PMID- 8720876 TI - The axonal transport of beta III-tubulin is altered in both branches of sensory axons after injury of the rat sciatic nerve. AB - We have analyzed the axonal transport of beta III-tubulin in the central (dorsal root) and peripheral (sciatic nerve) branches of sensory axons after injury of the sciatic nerve. Our finding that the relative amount of beta III-tubulin transported in slow component b (SCb) is increased in both axonal branches does not support the generally accepted hypothesis that the transport of cytoskeletal proteins is altered in the peripheral, but not the central branch after injury of the sciatic nerve. PMID- 8720877 TI - Synaptic relationships between axon terminals from the mediodorsal thalamic nucleus and layer III pyramidal cells in the prelimbic cortex of the rat. AB - A combined study of anterograde axonal degeneration and Golgi electron microscopic technique was designed to examine the distribution and density of axon terminals from the mediodorsal thalamic nucleus (MD) over layer III pyramidal cells in the prelimbic cortex of the rat. The reconstructive analysis of serial ultrathin sections of gold-toned apical and basal dendrites of layer III pyramidal cells showed that degenerating thalamocortical axon terminals from MD formed asymmetrical synaptic contacts predominantly with dendritic spines of the identified basal dendrites as well as apical dendrites. There was little difference in the numerical density of thalamocortical synapses from MD per unit length of both apical and basal dendrites. PMID- 8720878 TI - A differential effect of APB on ON- and OFF-center ganglion cells in the dark adapted rabbit retina. AB - The glutamate analog, 2-amino-4-phosphonobutyric acid (APB) is a proven tool in exploring the retinal circuit; it has been shown to interfere specifically with the transmission from photoreceptor to depolarizing bipolar cell. Consequently, in photopic retinae, the application of APB disrupts the ON-channel leaving the OFF-channel undisturbed; on the other hand, in the scotopic state, APB application blocks all ganglion cell responses. In this paper, we will show that the ON- and OFF-channels have a differential sensitivity to application of APB. That is to say, APB blocks center responses in ON-ganglion cells at mean concentration of 22 +/- 5.1 microM (mean +/- standard error of the mean; n = 15) and in OFF-ganglion cells at mean concentration of 91 +/- 15.5 microM (n = 16). Since considerable data rule out direct effects of APB on ganglion cells, we hypothesize that this effect is due to a difference in the synaptic gain of ON and OFF pathways in the inner retina. PMID- 8720879 TI - Effect of neuronal NO synthase inhibition on the cerebral vasodilatory response to somatosensory stimulation. AB - Whether nitric oxide (NO) mediates--or not--the local cerebral blood flow (CBF) increases occurring during functional brain activation is still a controversial issue. In the present study, we sought to determine whether neuronal NO synthase is involved in the cerebrovascular response to activation of the trigeminal pathway in the rat. Local CBF was measured using the autoradiographic [14C]iodoantipyrine technique in control alpha-chloralose anesthetized rats and 30 min following administration of 7-nitroindazole (7-NI), an inhibitor of the neuronal NO synthase. Unilateral whiskers stroking increased local CBF in all six regions of the trigeminal pathway. Under 7-NI, CBF was slightly decreased and the vasodilatatory response to whisker stimulation was unaltered in the four trigeminal nuclei studied. In contrast, no significant vasodilatation was noted in the ventral posteromedial thalamic nucleus and somatosensory cortex. These results suggest that the neuronal NO synthase mediates the hyperemia associated with somatosensory activation in second order relay stations but not in the site of termination of primary afferents. PMID- 8720880 TI - Calretinin as a marker of specific neuronal subsets in primate substantia nigra and subthalamic nucleus. AB - Neurons immunoreactive for the calcium-binding protein calretinin (CR) were visualized in the substantia nigra (SN) pars compacta (SNc), reticulata (SNr) and lateralis (SNl), the ventral tegmental area (VTA) and the subthalamic nucleus (STN) of squirrel monkeys. The density of CR perikarya was high in VTA, moderate in SNc, low in SNr/SNl, and very low in STN. The SNc/VTA complex also displayed a CR-rich neuropil. Some CR perikarya and fibers occurred in specific sectors of STN. These data suggest that CR may play a role in specific neuronal subpopulations in all components of primate basal ganglia. PMID- 8720881 TI - Sympathetic sprouting reverses decreases in membrane-associated activity of protein kinase C following septohippocampal denervation of the rat hippocampus. AB - Hippocampal sympathetic ingrowth (HSI), a form of neuronal plasticity, is induced by medial septal lesions and consists of the sprouting of peripheral sympathetic fibers, arising from the superior cervical ganglion, into the dentate gyrus and CA3 region of the hippocampus. HSI has been previously shown to alter learned and spontaneous behaviors, phosphatidyl inositide hydrolysis, and the antagonist binding kinetics of both muscarinic cholinergic receptors and phorbol ester receptors. We now report that sympathetic sprouting reverses decreases in membrane-associated activity of protein kinase C (PKC) following septohippocampal denervation of the rat hippocampus. Further, no changes were found in alpha, beta or gamma PKC isoenzymes among experimental groups, suggesting that the group A PKC isoforms do not mediate the observed changes in activity and phorbol ester binding. PMID- 8720882 TI - Serotonin1A receptors are increased in postmortem prefrontal cortex in schizophrenia. AB - Serotonin-5-HT1A receptors were measured with [3H]8-hydroxy-2-(di-n propyl)aminotetralin ([3H]8-OH-DPAT) in postmortem prefrontal cortex of 12 pairs of subjects with schizophrenia and age-matched control subjects. The saturation binding isotherms of [3H]8-OH-DPAT revealed high- and low-affinity binding sites. The density (Bmax) of the high-affinity sites was significantly elevated by an average of 79% in subjects with schizophrenia. The dissociation constant (Kd) of the high-affinity sites in subjects with schizophrenia was not significantly different from the control subjects. These results provide further evidence for a role of the serotonergic system in the pathophysiology of schizophrenia. PMID- 8720883 TI - [Vascular endothelium: partner to polymorphonuclear leukocytes in the inflammatory process]. PMID- 8720884 TI - [The role of polymorphonuclear neutrophilic leukocytes in the pathogenesis of acute respiratory distress syndrome (ARDS)]. AB - Polymorphonuclear leukocytes (PM-NL) constitute the first line of defence in the protection of the host from invading microorganisms. PMNL also contribute to the removal of cellular debris from necrotic tissues during reparative processes. For these purposes PMNL are armed with highly efficient bactericidal mechanisms which, under certain pathophysiological conditions, can be turned against the host himself. A vast body of evidence indicates that PMNL are able to cause lung injury which may be followed by the development of acute respiratory distress syndrome (ARDS). Accordingly, in patients with ARDS blood concentrations of inflammatory activators of PMNL are elevated, cytotoxic mechanisms of PMNL are enhanced and sequestration of these cells has been demonstrated to be inversely proportional to gas exchange. The manifestation of ARDS in leukopenic patients, however, indicates the development of this clinical syndrome independently of the presence of PMNL. The ability to differentiate between PMNL-dependent and PMNL independent pathways in the pathogenesis of this syndrome is not only of theoretical interest but also of therapeutic significance. Since the patient's systemic inflammatory response may vary according to the stage and type of the underlying disease, an exact qualitative and quantitative analysis of PMNL functions may provide the rationale for new anti-inflammatory drug regimens aimed at modifying the host's response without increasing the risk of infection. PMID- 8720885 TI - [Early contributions from Erlangen to the theory and practice of ether and chloroform anesthesia. 1. Heyfelder's clinical trial with ether and chloroform]. AB - The era of modern anaesthesia in Germany began on January 24th, 1847. This day, professor in ordinary Johann Ferdinand Heyfelder anaesthetized a patient with sulphuric ether in the clinic of surgery and ophthalmology of the University of Erlangen. By March 17th, 1847, Heyfelder had performed 121 surgical procedures under ether. The operations in majority were teeth-extractions, and a few more complex operations such as the treatment of a harelip or of lip cancer or the resection of the shoulder joint. Heyfelder described in detail 108 of these inhalations in a little book entitled The experiments with sulphuric ether. This monograph published in March, 1847, represents one of the first complete dissertations on sulphuric ether in the German literature. In a special chapter he analyzed the development of various physiological and psychological parameters during etherization. Heyfelder also examined blood and urine of some etherized patients and reported that he did not find any important or specific alterations. In 1847, Heyfelder was probably the first to apply salt-ether in man. After 4 administrations he concluded that salt ether acted more quickly but shorter than sulphuric ether. Advantageous were its application without problems and ease of induction. Disadvantageous were its high volatility, its price and the difficulty of getting it in a pure form. From December, 1847, on Heyfelder started to use chloroform. He was now able to perform more major operations, for example, the total resection of the hip-joint. In his book The experiments with sulphuric ether, salt ether, and chloroform he describes a great number of anaesthetic administrations using these 3 agents. In his summary Heyfelder concluded, that chloroform was undoubtly superior to sulphuric ether mainly because it was a quicker acting and longer lasting agent and leads to deeper narcosis. Moreover its application was much easier for it needed no special apparatus. However, because of its great anaesthetic potency, Heyfelder particularly demanded great caution in the application of chloroform. Explicitely he expected an assistant for chloroformizations, whose only duty was to supervise the inhalations and the patient--a forerunner of the modern specialized anaesthesiologist. PMID- 8720886 TI - [Awakening from anesthesia and recovery of cognitive function after desflurane or isoflurane]. AB - Desflurane is a new volatile anaesthetic with an extremely low blood/gas partition coefficient of 0.42. This should provide a rapid recovery from anaesthesia. METHODS: We studied 100 adult patients, ASA class I or II, undergoing elective orthopaedic surgery randomly assigned to anaesthesia with desflurane (n = 50) or isoflurane (n = 50) supplemented by nitrous oxide in oxygen. Clorazepat was given for premedication, fentanyl and thiopental for induction of anaesthesia, followed by maintenance with desflurane or isoflurane as clinically appropriate. Emergence from anaesthesia was measured as well as return of cognitive functions (extended Aldrete score, digit symbol substitution test, and visual analogue scales [VAS]). RESULTS: While the demographic characteristics and administrated doses of fentanyl and thiopental were comparable, the recovery profiles in both groups were different. After discontinuation of the volatile anaesthetics, times to extubation and ability to follow simple commands were significantly shorter after desflurane than after isoflurane. Extended Aldrete scores, estimation of the patients' physical condition, results of the digit symbol substitution test, measuring cognitive functions, and rates of drowsiness and weakness on VAS showed better recovery with less impairment of cognitive function in the desflurane group than in isoflurane patients even 120 min after anaesthesia. VAS pain scores and doses of analgesic drugs given within the first 2 postoperative hours, however, showed no significant differences. Desflurane patients were also judged fit for discharge from the recovery room significantly faster. CONCLUSIONS: Our results demonstrate that desflurane anaesthesia, even when supplemented by premedication, intraoperative opioids, and nitrous oxide may offer clinical advantages over isoflurane as far as the post-anaesthetic recovery profile is concerned. PMID- 8720887 TI - [Incidence and quality of dreaming during anesthesia with propofol in comparison with enflurane]. AB - Since the introduction of propofol into clinical practice, dreaming during general anaesthesia has been frequently reported with this drug. We investigated the incidence and character of these dreams with regard to distinction from intraoperative awareness and in comparison with habitual dreams or dreams during general anaesthesia with enflurane. METHOD: A total of 230 patients (ASA I-II) undergoing minor elective surgery were randomly assigned to one of two groups. Both groups received the same premedication (1 mg flunitrazepam) the previous evening. After induction using etomidate 2 mg/kg, anaesthesia in group I was maintained using 0.8%-1.5% enflurane supplemented in the sense of "balanced anaesthesia" with fentanyl, vecuronium, and nitrous oxide. Anaesthesia in group II was induced with propofol 2 mg/kg. After a saturation phase of approximately 10 mg/kg.h, 5-6 mg/kg.h was administered for maintenance supplemented by fentanyl, Vecuronium, and N2O as in group I. Interviews were conducted preoperatively, just after recovery from anaesthesia, and 24 h later. We inquired about preoperative fears or concerns, sensory perceptions, and the incidence and character (pleasant, unpleasant, indifferent) of habitual dreams. After recovery the patients were asked if they had had any conscious perceptions, whether they believe them to be dreams or reality, and repeated the preoperative questions as to common feelings and the character of sensory perceptions. RESULTS: The two groups were not entirely comparable due to the distribution of sex and age, but we did not find any differences in preoperative grades of fear and incidence of dreams. Of the propofol patients, 60% dreamed intraoperatively versus 11% in the enflurane group. No evidence of any intraoperative awareness was found. Because of the low incidence of dreams in the enflurane group, a comparison of detailed information between the groups could not be provided. Except for 1, all patients in the propofol group had very pleasant dreams compared to their habitual dreams (4.5% in the enflurane group). One day after surgery 37% of all patients in the propofol group remembered having dreamed intraoperatively. We did not found any correlation to the incidence of habitual dreaming, preoperative fear, age, sex, duration of anaesthesia, or total dose of propofol. Furthermore, intraoperative dreams differed from habitual dreams in character and content. CONCLUSIONS: We conclude that sensory experiences during propofol anaesthesia are not stimulus related perceptions or awareness, but dreams similar to normal ones. In view of the absence of clear signs of stimulus-related response, we presume that dreaming is a specific side effect of propofol. Because of the absence of difficulties, dreaming during propofol anaesthesia appears not to be a disadvantage with regard to the quality of anaesthesia. PMID- 8720888 TI - [Measurement of expired alcohol concentrations with a new electrochemical sensor. A model investigation to determine interference with volatile anesthetics and clinical application]. AB - Absorption of irrigating fluid in transurethral prostatic resection (TURP) and percutaneous nephrolitholapaxy (PNL) into veins or delayed absorption due to fluid extravasation may result in a TURP syndrome. The measurement of end-tidal breath alcohol concentration (et AC) as a monitor of absorption of irrigating fluid labelled with 2% ethanol is limited by the disturbance of infrared sensors by volatile anaesthetics and nitrous oxide (N2O) (Fig. 2). An electrochemical sensor is acceptable for this method. The aim of the present study was the evaluation of breath alcohol measurements using an electrochemical sensor device (Alcomed 3010, Envitec). The stability of the sensor in the presence of volatile anaesthetics was examined using a lung model. In a clinical investigation, the device was then applied to spontaneously breathing or mechanically ventilated patients inhaling volatile anaesthetics during endoscopic urological surgery. METHOD: A two-chamber lung model filled with water for performing noninvasive measurements at the mouth of a patient has already been introduced by Brunner et al. (Fig. 1). With the addition of different amounts of ethanol to the temperature-controlled water, a constant ethanol concentration is achievable in the air above the water that is dependent on adjustments of the ventilator. Increasing concentrations of volatile anaesthetics (isoflurane, enflurane, halothane, and sevoflurane) were added to the fresh gas flow (2 l O2/3 l N2O) and etACs were measured using the manually triggered self-absorbent electrochemical sensor. First, regression equations were established between breath alcohol concentrations and increased volatile anaesthetic concentrations. Regression equations were then established between end-tidal anaesthetic gas concentrations and vaporizer adjustments in order to rule out an influence of ethanol on the anaesthetic gas monitor Ultima V (Datex). In the clinical investigation, 53 intubated and ventilated patients (33 undergoing PNL, 20 undergoing TURP) and 48 patients breathing spontaneously (32 with inhalation anaesthesia, 16 with spinal anaesthesia) were investigated. The etAC was measured with the Alcomed 3010 and compared with gas-chromatographically registered blood alcohol concentrations (BAC). The study had previously been approved by the Ethical Committee of the Medical University of Luebeck. Patients with liver disease and a history of toxic abuse were excluded. Only one value per patient (maximum BAC) was included in the statistics in order to avoid a cluster effect. RESULTS: The lung model experiments demonstrated that the measurement of etAC with an electrochemical sensor is free of interference by volatile anaesthetics (Table 1). The slope of the regression between the measured alcohol concentration and increased concentrations of anaesthetics did not differ significantly from baseline values. The measurement of end-tidal anaesthetic concentrations was not significantly different from vaporizer adjustments in the presence of increased alcohol concentrations (Table 2). During the clinical investigation, a regression between etAC and BAC was determined for both groups. For the group of patients breathing spontaneously, the correlation coefficient was 0.961 and the regression equation revealed etAC = 0.5677*BAC-0.1303 (Fig. 5). However, in the group of ventilated patients a biphasic course was shown that was dependent on BAC (Fig. 6). At BAC < 0.4%, a similar correlation (r = 0.856) to the spontaneously breathing group could be seen (regression equation: etAC = 0.617*BAC-0.020). Above 0.4% BAC there was no acceptable correlation (r = 0.444, regression equation: etAC = 0.202*BAC+0.104). CONCLUSIONS: The tested electrochemical sensor does not interfere with volatile anaesthetics and N2O as demonstrated by a lung model. There is a good correlation between etAC and BAC measurements in patients breathing spontaneously with special regard to the slope of the regression (s = 0.57). PMID- 8720889 TI - [Are emergency physicians' diagnoses accurate?]. AB - METHODS AND AIM OF THE STUDY: The on-scene performance during all missions of the emergency physician-operated rescue helicopter and mobile intensive care unit based at a large-city hospital over a period of 1 year was retrospectively analysed; 2,254 hospital discharge reports were available (92% of the patients treated by the emergency physicians [n = 2,493]). The following parameters were investigated: reliability of the primary diagnosis established by the emergency physician (by comparison with the discharge diagnoses); initial on-scene therapeutic measures; means of transportation (with or without accompanying emergency physician); and level of care of the target hospital. RESULTS: The most common reasons for a mission were cardiopulmonary diseases (55%), neurological disorders (18%), and traumatic events (7%). The diagnoses, therapeutic measures, and mode of transportation were correct in 2,033 (90%) patients with a discharge report. Severe errors of assessment by the emergency physician were identified in 73 patients (3%): life-threatening conditions were not recognised and/or grossly incorrect therapeutic measures were taken and/or the chosen means of transportation was unsuitable. Relative errors in assessment occurred in 4% (n = 83): the most crucial diagnosis was not made, but the patient was escorted by the emergency physician (without therapeutic errors) to a suitable hospital. In 3% (n = 65) of the cases, the patient's condition was overestimated by the emergency physician as suggested by the obviously exaggerated on-scene therapy. Underestimations of the severity were most common in patients with cardiopulmonary diseases and increased in frequency and severity with increasing age and the presence of a concomitant neurologic deficit. Underestimations of a severe condition in younger patients were extremely rare; overestimations of the severity and consequent overtreatment were particularly common in traumatised patients independent of age. CONCLUSIONS: In the context of quality management measures, a careful evaluation of on-scene diagnoses, therapeutic measures, and decisions made by the emergency physician is a suitable procedure for identifying systematic errors. A high percentage of correct diagnoses and therapy at the emergency site can only be ensured by clinically experienced physicians who constantly deal with patients with acutely life-threatening conditions. PMID- 8720890 TI - [Anesthesia and intensive care management of severely burned children of Jehovah's Witnesses]. AB - A 3.5-year-old girl suffered from a thermal injury affecting 37% of the body surface area. The parents, being Jehovah's witnesses, refused permission for their child to receive blood transfusions. As the haemoglobin level was only 7.5% and a necrectomy was planned, the patient was likely to need blood transfusions. Indications for transfusion were defined as clinical signs of hypoxia and/or cardiovascular instability. A judicial declaration was proposed. Hb decreased during the therapy. To stimulate the erythropoiesis erythropoietin and iron were administered. During the clinical course the anaemia worsened. First, a conservative treatment with polyvidoniodine ointment for tanning was started, to avoid an operation during the acute phase after the injury, as in this case it was thought a blood transfusion would definitely be necessary. On the 19th day after the injury a necrectomy of 10% of the body surface was necessary because of fever and leucocytosis not responding to antibiotics. The most likely cause of the symptoms was an infection of the burned area. Hb was 4.6 g/dl%. General anaesthesia was performed with midazolam, ketamine and vecuronium and mechanical ventilation. No blood transfusion was required during the operation. Vital signs were stable during the preoperative period, during anaesthesia and following the operation. There were no signs of tissue hypoxia. Peripheral oxygen saturation ranged between 98% and 100%, lactate and arterial blood gas samples were normal, and the child was awake and cooperative before and after anaesthesia. The lowest Hb was 3.3 g/dl on the 22th day after injury (3rd postoperative day). In this phase the patient was still playing and riding a tricycle. On the 45th day after injury the child was discharged home with Hb of 10.9 g/dl and reticulocytosis of 33%. PMID- 8720891 TI - [Treatment reduction in intensive care. "Allowing the patient to die" by conscious withdrawal of medical procedures]. AB - The conversion of an "attempt to treat" to "prolongation of dying" represents an important problem in modern intensive care. If the actual or presumed will of the patient is unknown, the physician has to decide about the extent of treatment in a paternalistic manner. In these difficult decisions the physician has to consider prognosis, and certainty of prognosis and has to carefully balance between the right to live and the right to die. This decision about the extent of therapy is a very personal medical activity and can be taken off the physician's shoulders by nobody. Consultation with other physicians involved, relatives, nurses and clergy, however, is mandatory, as a joint decision should be sought. If the situation is hopeless and further medical interventions are futile, then allowing the patient to die by therapy reductions is not only a possibility but a mandatory act of humanity. In that case it does not matter whether new treatment modalities are abandoned or whether already instituted medical measures are withdrawn. In clinical practice, however, the "fine tuning" of therapy reduction has to be tailored to the individual case and largely depends on prognostic certainty. PMID- 8720892 TI - [Remarks on the letter of U. Schult-Sasse. Manufacturer warning for the routine administration of succinylcholine in children]. PMID- 8720893 TI - [Challenge and change. 68th Congress of the American Heart Association. Anaheim, California, November, 1995]. PMID- 8720894 TI - [Candida infections in intensive care]. PMID- 8720895 TI - IgG1--as the only subclass of human serum IgG--spontaneously undergoes O2(-) induced, noncovalent self-aggregation upon storage at room temperature. AB - An apparent gradual decrease of IgG1 serum levels of up to 40% occurs within 48 h of storage at room temperature. The effect does not concern any other IgG subclass, and is more pronounced in sera of smokers. A linear correlation was found between the extent of this "storage effect" and the initial concentration of IgG1, which rules out an enzymatic process following Michaelis-Menten kinetics. PAGE and Western blots of density gradient separated serum proteins revealed the presence of noncovalent self aggregates of IgG1 in stored sera. Addition of superoxide dismutase prevented both the formation of aggregates as well as the decay of IgG1 values. It is concluded that the instability of IgG1 is due to an enhanced propensity of this molecule to form self-aggregates, whereby O2(-)-radicals play a functional role. This mechanism, however, is not relevant to a previously detected selective decrease of relative IgG1 levels in sera of patients afflicted with malignant diseases of various tissue origin. PMID- 8720896 TI - Platelet-neutrophil interaction and superoxide anion generation: involvement of purine nucleotides. AB - Stimulated platelets secrete a variety of physiologically active substances that affect many neutrophil functions. We have examined the capacity of platelets to modulate superoxide anion generation by neutrophils. The amounts of superoxide anion produced by neutrophils in the presence of platelets were markedly enhanced when platelet-neutrophil coincubations were stimulated with agents that simultaneously activate both cell types, as the calcium ionophore A23187 and sodium arachidonate. This effect was dependent upon the number of platelets added to the incubation media and was not affected by inhibitors of arachidonic acid pathway or by preincubation of platelets with an antibody anti-P-selectin. The hypothesis of an involvement of purine nucleotides released by platelets during aggregation on the observed effect of enhancement of superoxide anion generation by neutrophils was then tested. Experimental evidence indicates that platelets release, during A23187-induced aggregation, amounts of ATP that are of the same order (5-10 microM) of those demonstrated to enhance superoxide anion generation by neutrophils. In addition, platelet lysates mimicked the effect of intact platelets in enhancing superoxide anion generation by A23187 stimulated neutrophils. Interestingly, at variance with the results obtained with intact platelets and platelet lysates, supernatants of thrombin-stimulated platelets did not increase O2.- by neutrophils. The enhancing effect of these supernatants was, however, restored when platelets were preincubated with an antibody anti P selectin. These data indicate that platelets, through the release of purine nucleotides, enhance superoxide generation by neutrophils, thus increasing the cytotoxic potential of these cells. PMID- 8720897 TI - The effect of dietary supplements on gene expression in mice tissues. AB - Exposure of living organisms to various environmental stresses induces the synthesis of so-called shock/stress proteins; many of them can provide either immediate stress protection or participate in cellular repair processes. In the present study we focused our attention on the potential effect of dietary vitamins and microelements with antioxidant properties on stress protein gene expression. The analysis of gene expression in tissues of antioxidant-fed mice shows hsp-70 gene overexpression in liver and brain, but not in spleen and lung. Heat shock significantly induces gene expression that is less pronounced in antioxidant-fed animals in all analyzed tissues. Under conditions of oxidative stress, accumulation of lipid peroxidation products in liver homogenates is partially suppressed in mice subjected to heat shock, and significantly inhibited in antioxidant-fed mice and in antioxidant-fed mice subjected to heat shock. The glutathione content in liver homogenates of antioxidant-fed mice is higher than in the control group. Heat shock decreases the level of endogenous glutathione in both groups of animals, but it is still higher in the liver homogenate of antioxidant-fed mice. Thus, dietary supplements can modify gene expression induced by heat shock in vivo and protect rat tissues against oxidative stress by enhancing the level of endogenous antioxidants and inducing hsp-70 gene expression. PMID- 8720898 TI - Sod and catalase inactivation by singlet oxygen and peroxyl radicals. AB - Both superoxide dismutase and catalase are readily deactivated by singlet oxygen and by the radicals produced in the pyrolysis of 2,2'-azo-bis-(2-amidinpropano) under aerobic conditions. The rate constant for the loss of enzymatic activity induced by singlet oxygen are 3.9 x 10(7) and 2.5 x 10(7) M-1 sec-1 for SOD and catalase, respectively. The similarity between these values implies that in systems where SOD and catalase are exposed to similar singlet oxygen concentrations, it can be expected a parallel inactivation of both enzymes. The inactivation of both enzymes by the radicals produced by 2,2'-azo-bis-(2 amidinopropane) pyrolysis under aerobic conditions follows a first-order kinetics at low enzyme concentrations and a zero-order kinetics at higher concentrations. Although at low enzymatic concentrations the rate of inactivation of both enzymes is similar, this results from a compensation of effects because there are wide differences in the reactivity of both enzymes towards peroxyalkyl radicals. Catalase is considerably more reactive, but a large number of protein/radical reactive interactions are needed to inactivate one enzyme. On the other hand, the reactivity of SOD is smaller, but the average enzyme activity decreases by nearly 20% in each SOD/radical reactive interaction. PMID- 8720900 TI - Lens epithelial cell apoptosis is an early event in the development of UVB induced cataract. AB - Epidemiological and experimental studies have revealed that exposure to UV can induce cataractogenesis. To investigate the mechanism of this induction, viability of the lens epithelial cells from UVB-treated rat lenses were examined. Irradiation of the cultured rat lenses with 8 J/s/m2 UVB for 60 min triggers lens epithelial cell apoptosis as determined by terminal deoxyribonucleotide transferase (TdT) labeling and DNA fragmentation assays. The apoptotic lens epithelial cells were initially found in the equatorial region and then quickly appeared in both equatorial and central regions. The percentage of apoptotic cells continuously increased during the postirradiation incubation. After a 5-h post-UVB incubation, more than 50% of the lens epithelial cells were apoptotic. By 24 h, all of the lens epithelial cells in the irradiated lenses were dead through apoptosis. Associated with this apoptotic process is a large upregulation of the proto-oncogene, c-fos. Opacification appears to follow the death of lens epithelial cells occurring first in the equatorial region and then in the central area. This is also true of classical cataract parameters such as non-protein thiol and wet weight, which are significantly modified only after appreciable epithelial cell apoptosis. Together, these results suggest that the rapid apoptotic death of the lens epithelial cells induced by UVB initiates cataract development. PMID- 8720899 TI - The prooxidant effect of 5-aminolevulinic acid in the brain tissue of rats: implications in neuropsychiatric manifestations in porphyrias. AB - 5-Aminolevulinic acid (ALA), a heme precursor accumulated during the clinical expression of acute intermittent porphyria, lead poisoning, and tyrosinosis, has been hypothesized to act as an endogenous source of oxyradicals. We now report oxidative effects on brain tissue of rats submitted to ALA treatment. Upon acute treatment (40 mg/kg body weight) increased total nonheme iron in the cortex (20%) was observed. After prolonged ALA administration (40 mg/kg body weight on alternate days during 2 weeks), the following indicators of oxidative stress were found to be significantly increased: CuZnSOD activity (67%) in total brain homogenate, total iron (68%) and ferritin (71%) in the cortex, ferritin in striatum (44%), protein carbonyls in homogenate of cerebral cortex (threefold) and 45Ca2+ uptake by cortical synaptosomes (45%). In addition, synaptic membranes prepared from whole brain assayed with the radioligand 3H-muscimol, revealed increased Kd values (twofold) of the high-affinity GABAergic receptor binding and formation of protein carbonyl groups, thiobarbituric acid reactive products, and conjugated dienes. In vitro, ALA produced similar effects upon the high affinity 3H-muscimol binding. No apparent alteration of either dopaminergic or serotonergic [3H]-ligand binding was observed. These results argue in favor of ALA-triggered oxidative stress in brain accompanied by iron metabolism alterations and GABAergic receptor damage, which may be implicated in the neuropsychiatric manifestations of the aforementioned porphyrias. PMID- 8720901 TI - Nuclear DNA damage during NAD(P)H oxidation by membrane redox chains. AB - Nuclear DNA damage, as the result of active oxygen formation by NAD(P)H-dependent redox chains, was studied. Isolated rat liver nuclei were incubated in the presence of NAD(P)H and iron chelators. Nuclear DNA damage was analyzed by electrophoresis in alkaline agarose. DNA damage after the addition of electron donors alone or with FeCl3 or DFO-Fe3+ was not visualized. Dramatic decay of high molecular weight DNA was observed with EDTA-Fe3+ or DTPA-Fe3+ in the presence of NAD(P)H. SOD did not prevent DNA damage, whereas catalase was protective. DNA damage was revealed after the addition of cumene hydroperoxide with EDTA-Fe3+, and it was sharply increased in the presence of NADPH. It is suggested that alkoxyl radicals in addition to hydroxyl radicals are involved in DNA damage during NAD(P)H oxidation in the presence of iron chelators, which can be reduced by membrane redox chains. PMID- 8720902 TI - Depletion of cellular iron by bps and ascorbate: effect on toxicity of adriamycin. AB - A new method was developed that reduces the intracellular iron content of cells grown in serum-containing culture without involving the significant uptake of iron-chelating agents into cells. Negatively charged bathophenanthrolinedisulfonate (BPS), together with ascorbate, caused cells to lose much of their cellular iron without causing much depression in HL-60 or H9c2 (2-1) cell proliferation over a 48-h period. When added to serum supplemented RPMI-1640 culture media, BPS and ascorbate efficiently reduced and competed for iron in Fe(III) transferrin to form Fe(II)(BPS)3. The reaction also occurred with purified human iron-transferrin. When cells were incubated with growth medium containing serum that had been treated with BPS and ascorbate for 24 h, little or no BPS2- or Fe(II)(BPS)(4-)3 entered the cells, according to direct measurements and in agreement with the highly unfavorable 1-octanol/water partition coefficients for these molecules. However, iron was mobilized out of both cell types. After 24 h incubation of cells in this medium, there was no change in the activities of catalase and superoxide dismutase, or in the concentration of glutathione. Glutathione peroxidase was elevated 9%. Using HL-60 and H9c2 (2-1) cells made iron deficient with BPS and ascorbate, HL-60 cells grown in defined growth media in the absence of iron-pyridoxal isonicotinoyl hydrazone, or Euglena gracilis cells maintained in a defined medium that was rigorously depleted of iron, it was shown that the cytotoxicity of adriamycin is markedly dependent on the presence of iron in each type of cell. Similar results were obtained when HL 60 cells were grown in RPMI-1640 culture medium and serum that had been incubated for 24 h in BPS and ascorbate and then chromatographed over a Bio-Rad desalting column to remove small molecules including BPS, ascorbate, and Fe(II)(BPS)3. PMID- 8720903 TI - Structural aspects of antioxidant activity of flavonoids. AB - Flavonoids, a group of naturally occurring antioxidants and iron chelators, might be used as cardioprotective agents in doxorubicin-induced cardiotoxicity, which is believed to be caused by the formation of oxygen free radicals. To investigate the underlying molecular mechanism, we tested a large group of flavonoids from all major structural subclasses on their ability to inhibit doxorubicin (enzymatically)-induced and Fe2+/ascorbate (nonenzymatically)-induced microsomal lipid peroxidation (LPO) and to chelate Fe2+. In addition, we measured half peak oxidation potentials (Ep/2). LPO inhibition data gave a good qualitative correlation with the oxidation potentials. Most flavonoids tested chelated Fe2+, but there were large differences in the chelating capacity. For good scavenging activity, a catechol moiety on ring B is required. The 3-OH moiety can function as a chelation site and can also be oxidized. The 3-OH group in combination with a C2 C3 double bond, increases the scavenging activity. Fe2+ chelation only plays a role in the LPO inhibition by less active scavengers. Chelation can then raise the activity to the level of the most active scavengers, possibly by site specific scavenging. It can be concluded that Ep/2 values and iron chelating activity can almost completely describe the LPO inhibiting behaviour of the flavonoids. PMID- 8720904 TI - Peroxynitrite-induced cardiac myocyte injury. AB - The effects of peroxynitrite (ONOO-) on cultured cardiac myocytes were examined by simultaneous measurements of intracellular Ca2+ ([Ca2+]i) and contractile function. On exposure to 0.2 mM ONOO-, [Ca2+]i increased to beyond the systolic level within 5 min with a concomitant decrease in spontaneous contraction of myocytes followed by complete arrest. Addition of a L-type Ca2+ channel blocker or removal of extracellular Ca2+ prevented the ONOO(-)-induced increase in [Ca2+]i, indicating that the increase in [Ca2+]i was caused by the enhanced influx of Ca2+ through the plasma membrane and not by the enhanced release from sarcoplasmic reticulum (SR). Plasma membrane fluidity and concentration of the thiobarbiturate acid-reactive substance (TBARS) in the cells remained unchanged by the ONOO- treatment. The complete cessation of contraction of myocytes persisted even under the massive increase in [Ca2+]i, which was induced by an additional saponin (5 microM) treatment. In conclusion, ONOO- increases [Ca2+]i in myocytes through disturbance of Ca2+ transport systems in the plasma membrane and impairs contractile protein. PMID- 8720905 TI - On the role of lipid peroxidation in the pathogenesis of liver damage induced by long-standing cholestasis. AB - Previous studies have suggested a possible involvement of free radical reactions in the pathogenesis of cholestatic liver injury as well as in the modulation of hepatic fibrogenesis. In this study we investigated whether lipid peroxidation is involved in the development of chronic liver damage induced by long-standing cholestasis. For this purpose we have used the rat model of bile duct ligation (BDL), which leads to liver fibrosis and cirrhosis. Using this model we observed that the development of chronic liver damage was associated with the onset of lipid peroxidation, as pointed out by detection of carbonyl compounds, 4 hydroxynonenal (HNE) and malondialdehyde (MDA), in BDL livers and of fluorescent adducts between MDA and serum proteins. Lipid peroxidation was a relatively late event (starting after 1-2 weeks of BDL) and was unrelated to the early development of liver necrosis and cholestasis (already evident after 72 h after BDL). A positive significant linear correlation between the kinetic of infiltration of neutrophils and of a monocyte/macrophage population in BDL livers and MDA and HNE generation in the same organs is presented, indicating a close link between lipid peroxidation and the activation of inflammatory cells. We also observed that a positive linear correlation exists between collagen deposition in these livers and hepatic production of MDA and HNE. This event, which is accompanied by an increase in the number of fat storing cells (FSC, the cells that produce collagen in fibrotic liver), suggests that lipid peroxidation in this model may contribute to stimulate collagen synthesis by proliferating FSC. PMID- 8720906 TI - Location-dependent artifact for no measurement using multiwell plates. AB - The Griess reaction is widely used to measure the cellular production of NO by detecting the supernatant levels of nitrite. Ordinarily, background levels of nitrite in the media are subtracted from the levels of nitrite produced by the cells by preparing a "blank" during the determination of the standard curve. Although this method is adequate for most experimental conditions, it cannot be used when cell supernatants are collected from multiwell dishes, particularly when low amounts of NO are produced and when long incubation periods are required to induce NO production. Our data show that a highly variable level of nitrite is found in the absence of cells in the media from wells at the edges of the 96-well plate while media from interior wells shows no detectable nitrite accumulation. The most likely source of this noncellular NO is from nitric oxides (NOx) found in the ambient air and reduction of air exchange or regulation of the gaseous environment eliminates this "border effect." PMID- 8720907 TI - Coulometric detection in high-performance liquid chromatographic analysis of cholesteryl ester hydroperoxides. AB - A highly sensitive and simple method for the determination of cholesteryl ester hydroperoxides (ChE-OOH) was developed using high-performance liquid chromatography (HPLC) with coulometric electrochemical detection. The lowest detectable level by this technique was 2 pmol for cholesteryl linoleate hydroperoxides at the signal-to-noise ratio of 3. This method was applied to the determination of ChE-OOH presumably present in human plasma. Although ChE-OOH could not be detected, the ChE-OOH level in the fluid was estimated to be less than 27 nM. It was found that the extraction efficiency of an internal standard, cholesteryl nervonate, was decreased by lowering its amount spiked to the plasma. The concentration of ChE-OOH in human plasma and plasma lipoprotein, which were peroxidized with a radical initiator in vitro, could be determined by use of this standard. HPLC-coulometric technique is, therefore, useful to measure the peroxidation of plasma lipids in vitro. PMID- 8720908 TI - Reactions of nitric oxide and peroxynitrite with organic molecules and ferrihorseradish peroxidase: interference with the determination of hydrogen peroxide. AB - Endothelial, inflammatory, and other cell types, in addition to forming reactive oxygen species, under proper stimulation release free radicals such as nitric oxide (.NO) and strong oxidants such as peroxynitrite (ONOO-), which is the product of the reaction of nitric oxide with superoxide. Several methods for the detection of H2O2 are based on the ferrihorseradish peroxidase catalyzed oxidation of organic molecules. We investigated the reactions of nitric oxide and peroxynitrite with organic molecules as well as with ferrihorseradish peroxidase and examined the potential interference with the detection of H2O2. Peroxynitrite at low concentrations (0-10 microM) induced a concentration-dependent oxidation of 1,2 phenylenediamine, 3-3' dimethoxybenzidine (o-dianisidine) and para hydroxyphenylacetic acid (pHPA). With the exception of pHPA, the oxidation of the above compounds by peroxynitrite was not affected by the presence of ferrihorseradish peroxidase. The yield of HPA-dimmer, the oxidation product of pHPA by peroxynitrite, was decreased because ferrihorseradish peroxidase catalyzed the formation of a different product, 3-nitro-HPA. Nitrogen oxides, formed by the reaction of nitric oxide with oxygen, oxidized the aromatic amines o-phenylenediamine and o-dianisidine. A 10-fold excess of nitric oxide over H2O2 decreased the yield of pHPA and dihydrorhodamine 123 (DHR123) by 58 and 72%, respectively, as compared to H2O2 plus ferrihorseradish peroxidase. The inhibition of pHPA oxidation by nitric oxide was in part to the reaction of nitric oxide with compound I and compound II and in part due to the reaction with the phenoxyl radical. These data suggest that the simultaneous generation of nitric oxide and peroxynitrite can interfere with the detection of H2O2. The overestimation or underestimation of the H2O2 detected is dependent upon the organic molecule utilized for detection and by the relative rate of nitric oxide, superoxide, and peroxynitrite generation. PMID- 8720909 TI - Protective effects of the K+ ATP channel opener, aprikalim, against free radicals in isolated rabbit hearts. AB - Aprikalim, a K+ ATP channel opener, is a potent vasodilator with demonstrated cardioprotective properties against ischemia/reperfusion injury. It is still unknown if K+ ATP channel openers exert their beneficial effects via interaction with oxygen-derived free radicals. Therefore, we investigated the cardioprotective effects of aprikalim against oxygen-derived free radicals. Isolated rabbit hearts were perfused at constant pressure (85 cm H2O) or constant flow (30-35 ml/min). Heart rate, left ventricular developed pressure (LVDP), and either coronary flow or coronary perfusion pressure (CPP) were monitored. Free radicals were produced by electrolysis of the perfusate (0.6 mA, direct current), and 10 microM aprikalim was infused before and after exposure to free radicals. In the constant perfusion pressure experiments, 10 min of exposure to free radicals resulted in a significant reduction of heart rate (137 to 129 beats/min), LVDP (112 to 91 mmHg) and coronary flow (37 to 29 ml/min); coronary flow was more markedly impaired than contractile function. Acetylcholine-induced coronary dilation was also significantly attenuated in the presence of free radicals. After 30 min of recovery, both coronary flow and LVDP were still significantly decreased while acetylcholine-induced coronary dilation had fully recuperated. Aprikalim completely abated the coronary and cardiac depressant actions of free radicals. Constant flow experiments indicated that exposure to free radicals increased CPP (+40%, p < 0.05), an effect totally suppressed by aprikalim. These results demonstrate that aprikalim reverses the cardiodepressant actions of free radicals. The cardioprotection it afforded involves both contractile function and the coronary vasculature. Acetylcholine-induced coronary dilation was blunted by free radicals, an indication of complex interactions at the coronary endothelial level. PMID- 8720910 TI - Stress proteins and SH-groups in oxidant-induced cell damage after acute ethanol administration in rat. AB - It is generally accepted that lipid peroxides play an important role in the pathogenesis of ethanol-induced cellular injury and that free sulfhydryl groups are vital in cellular defense against endogenous or exogenous oxidants. It has been observed that oxidative stress induces the synthesis of the 70-kDa family of heat-shock proteins (HSPs). Furthermore, induction of HSPs represents an essential and highly conserved cellular response to a variety of stressful stimuli. In the present study, we measured the intracellular levels of HSP 70 proteins after administration of mild intoxicating and grossly intoxicating doses of ethanol to rats. Our results demonstrate that elevated doses of ethanol induce HSP in various brain areas, namely, cerebellum, hippocampus, and to a lesser extent, striatum or liver. Induction of HSP 70 protein was correlated with a marked depletion of intracellular bound thiols and a decrease in lipid peroxidation measured as MDA formation. These studies support the hypothesis that a redox mechanism may be involved in the heat-shock signal pathway. PMID- 8720912 TI - Reduced iron-associated antioxidants in premature newborns suffering intracerebral hemorrhage. AB - Oxygen radical injury may be a common pathogenic mechanism in several neonatal diseases. The term "oxygen radical disease of prematurity" has been proposed in the face of the greater incidence of intracerebral hemorrhage, bronchopulmonary dysplasia, and retinopathy in premature neonates. To test the hypothesis that overload with ionic iron due to decreased concentrations of iron-oxidizing and iron-binding proteins induces free radical damage in premature asphyxiated newborns suffering periventricular-intraventricular hemorrhage (PIVH), blood plasma of newborns with PIVH (n = 7) was compared with that of controls (n = 10) within the first 12 h of life. We found reduced transferrin (2.05 vs. 2.24 g/l; p < 0.05) and ceruloplasmin (89.9 vs. 126.3 mg/l; p < 0.01) levels and an increased transferrin saturation (54.2 vs. 38.4%; p < 0.05) in those newborns who later developed PIVH. These findings support the theory that iron-catalyzed lipid peroxidation of the brain during reoxygenation after perinatal asphyxia may be involved in the pathogenesis of PIVH. PMID- 8720911 TI - Evidence for involvement of multiple iron species in DNA single-strand scission by H2O2 in HL-60 cells. AB - Some of the properties of cellular iron species which react with H2O2 to cause DNA single-strand breaks in HL-60 cells were characterized in control cells and in cells made deficient of iron using 4,7-phenylsulfonyl-1,10-phenanthroline (bathophenanthroline disulfonic acid or BPS) and ascorbate. Single-strand breaks were measured using alkaline elution of DNA of cells treated at 4 degrees to minimize repair during treatment. Strand breakage in the presence of 10% serum was only 40% of that in the absence of serum. This effect was traced to reaction of H2O2 with metals, most likely iron, in serum. Dimethyl sulfoxide (Me2SO) inhibited a maximum of 65% of breaks in control cells. The diffusion distance from the site of generation of hydroxyl radicals to the site of reaction with DNA for the Me2SO-inhibitable fraction was 6.9 nm. There was no significant alteration in the fraction of Me2SO-inhibitable strand breaks or in diffusion distance in iron-deficient cells, though total strand breaks decreased by 70%. When the effect of extracellular iron in serum was taken into account, 60 microM orthophenanthroline (OP) inhibited a maximum of 85% of strand breaks. In cells pretreated with 60 microM OP, the Me2SO-inhibitable fraction of the remaining strand breaks decreased to 32%, while the diffusion distance decreased to 4.1 nm. These data indicate the existence of a number of different iron species, as characterized by overlapping but not coincidental inhibition by OP and Me2SO, and by differing diffusion distances. PMID- 8720913 TI - Quenching of reactive oxidative species by probucol and comparison with other antioxidants. AB - One-electron oxidation of the antiatherogenic and antiatherosclerotic drug probucol has been studied in relation to its activity as an antioxidant. Oxidation by triplet excited states of duroquinone and benzophenone, and by the inorganic radicals Br2.- and N3., lead to the formation of a transient absorption at 500 nm. This was identified by time-resolved resonance Raman spectroscopy as the phenoxyl radical from probucol, formed by hydrogen atom or electron plus proton loss from one of the phenolic groups of probucol. The reactivity of probucol with triplet duroquinone and triplet benzophenone, and as a quencher of singlet oxygen, was compared with the reactivities of other antioxidants (alpha tocopherol, palmitoyl ascorbic acid, dihydrolipoic acid and N-acetyl cysteine). In quenching of the triplet states the reactivity of probucol was comparable with that of alpha-tocopherol, whereas as a quencher of singlet oxygen probucol (k < 10(6) M-1 s-1) was less effective than alpha-tocopherol (k = 2.0 x 10(8) M-1 s-1) by more than two orders of magnitude. This difference in reactivity may allow the contribution of singlet oxygen towards oxidative stress to be quantified separately. PMID- 8720914 TI - Antioxidant properties of amidothionophosphates: novel antioxidant molecules. AB - This work describes the synthesis and characterization of a new family of antioxidants. The molecules have the same active group, but different oil-to water and octanol-to-water partition coefficients due to different substituents. Three new molecules were synthesized based on the chemical structure of the primary amide attached to a thiophosphate group forming an amidothionophosphate. The amidothionophosphate molecules were exposed to the oxidative stress of hydrogen peroxide and sodium hypochlorite, and the chemical changes following the exposure were monitored by 31P NMR. The reaction constants with the reactive oxygen species hydroxyl radical and superoxide radical were also calculated and found to be 1.5 x 10(9) M-1s-1 and 8.1 x 10(2) M-1s-1, respectively. To elucidate the ability of amidothionophosphates to act as antioxidants in protecting lipids and proteins, we examined damage prevention in bovine serum albumin, egg phosphatidylcholine liposomes, and lipid emulsions following oxidative stress. Amidothionophosphate showed unique protection properties in these models. In contrast to other antioxidant molecules (ascorbic acid, cysteine, and alpha tocopherol) the new group did not have any pro-oxidative effects as measured by oxygen consumption from buffer solutions containing amidothionophosphates and cupric sulfate as a source of redox-active metal ions. Amidothionophosphates reduced significantly and in a dose-dependent manner the oxidative burst in human neutrophils as measured by luminol-dependent chemiluminescence, and they also markedly depressed the killing of human fibroblasts by mixtures of glucose oxidase and streptolysin S. The toxicity of these molecules was tested by IP injection of doses up to 1000 mg/kg to white Sabra mice. No mortality was observed 30 d after administration of up to 500 mg/kg. PMID- 8720915 TI - The reduction of glutathione disulfide produced by t-butyl hydroperoxide in respiring mitochondria. AB - Factors affecting the reduction of GSSG by rat liver mitochondria after a t-butyl hydroperoxide-induced (t-BOOH) oxidative stress were studied. The amounts of ADP and mitochondrial protein were adjusted to consume less than 50% of the available oxygen during the 8-min experimental period. A 4-min treatment of mitochondria with 24 nmol t-BOOH/mg protein (60 microM) oxidized 91% of total glutathione. In the presence of glutamate/malate, succinate or ascorbate/N,N,N',N'-tetramethyl-p- phenylenediamine (TMPD) (state 4 respiration), 84, 84, and 28% of the GSSG formed during t-BOOH treatment was reduced after 4 min, respectively. A similar extent of reduction was seen during state 3 respiration (1.5 mM ADP) with glutamate/malate, but no reduction occurred during state 3 respiration with either succinate or ascorbate/TMPD. The succinate-supported reduction of GSSG was completely blocked by rotenone, antimycin A, carbonyl cyanide p trifluoromethoxyphenylhydrazone (FCCP), or 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU). In contrast, oligomycin potentiated GSSG reduction using either glutamate/malate or succinate as substrates. Rotenone partially blocked glutamate/malate-supported GSSG reduction. NADPH levels were altered in direct proportion to the effects on GSSG reduction. The current data indicate that the reduction of GSSG in oxidatively-stressed isolated rat liver mitochondria occurs most efficiently during state 4 respiration and is independent of ATP synthesis. Both transhydrogenation and the transmembrane proton gradient appear to be important in NADPH regeneration and consequent GSSG reduction. PMID- 8720916 TI - Superoxide anion and endothelial regulation of arterial tone. AB - Evidence continues to accumulate on the importance of paracrine substances formed in the endothelium in regulation of the vascular system. Mechanisms that govern the balance between relaxing and contracting factors are important for understanding the regulation of blood vessel tone in health and disease. Chemical antagonism between superoxide anions and nitric oxide has been recognized as a potentially important modulator of vascular reactivity as well as being a source of peroxynitrite, a potent oxidant. In several models of vascular diseases, impairment of endothelium-dependent relaxations and promotion of endothelium dependent contractions has been ascribed to increased production of superoxide anions. In pathologic conditions, increased production of superoxide anions may be responsible for an impairment of balance between relaxing and contracting factors favoring an increase in arterial tone. In this review the role of superoxide anions in modulation of endothelial mechanisms responsible for regulation of arterial tone will be discussed. PMID- 8720917 TI - Enhanced levels of chemiluminescence and platelet activating factor in urease positive gastric ulcers. AB - Helicobacter pylori are believed to play an important role in the formation of gastric ulcer in a syndrome characterized by a high urease activity. On the other hand, the production of oxygen radicals and platelet activating factor (PAF) is enhanced in gastric ulcers. The present study is designed to investigate the relationship between the different aspects of gastric mucosal injury, urease activity, oxygen radical production, and PAF content in gastric specimens. Biopsy specimens taken from 35 gastric ulcer patients were studied. Urease activity was detected by a rapid urease test (CLO). Oxygen radical production was measured as a value of luminol-dependent chemiluminescence (ChL) and PAF content was determined by radioimmunoassay in the biopsy samples. The CLO-positive rate was significantly higher in the gastric ulcer group in comparison with that in controls. ChL values and PAF content were significantly increased in gastric ulcers, especially in CLO-positive specimens. The CLO-positive rate, ChL values, and PAF content were also found to be increased at a distant site beyond the ulcer lesions. During the course of macroscopic ulcer healing of CLO-positive cases, the CLO positive level and the ChL values were not significantly decreased, although PAF content was significantly lower. Enhanced oxygen radical and PAF production were observed not only in the ulcer region but also at a distant site from the ulcer in the urease-positive gastric mucosa. The persistent enhancement of ChL values during the healing stage of urease-positive gastric ulcers suggests its involvement in the recurrence of gastric ulcers. PMID- 8720918 TI - Effect of 4-Hydroxynonenal on cell cycle progression and expression of differentiation-associated antigens in HL-60 cells. AB - 4-Hydroxynonenal (HNE) is a highly reactive aldehyde produced by lipid peroxidation of cellular membranes that inhibits growth and induces differentiation in HL-60 cells. Its mechanisms of action were investigated by analyzing the cell cycle distribution and the appearance of differentiated phenotypes in HL-60 cells. Data obtained by exposing cells to DMSO for 7.5 h (same time as for HNE treatment) or for the whole length of the experiments (5 d) were used for comparison. HNE induced a marked increase in the proportion of G0/G1 cells after 1 and 2 d. The brief DMSO treatment did not affect the distribution, whereas continuous exposure led to a progressive accumulation of cells in G0/G1 (maximal at day 5). The proportion of phagocytic cells gradually increased in HNE-treated and DMSO long-exposed cultures from day 2 and peaked at day 5 (35 and 63%, respectively), whereas the effect of the brief DMSO treatment was negligible. The expression of CD11b and CD67 increased in cells treated with HNE or continuously exposed to DMSO, whereas CD36 was expressed at low levels on both treatments. These results indicate that the pathway of the granulocytic differentiation induced by HNE in HL-60 cells differs from that of DMSO: with HNE, growth inhibition precedes the onset of differentiation, whereas in DMSO treated cells the two processes are chronologically associated. PMID- 8720919 TI - Low antioxidant enzyme gene expression in pancreatic islets compared with various other mouse tissues. AB - Using a sensitive Northern blot hybridization technique, gene expression of superoxide dismutase (SOD), catalase, and glutathione peroxidase was studied in pancreatic islets and for comparison in various other mouse tissues (liver, kidney, brain, lung, skeletal muscle, heart muscle, adrenal gland, and pituitary gland). Gene expression of the antioxidant enzymes was usually in the range of +/ 50% of that in the liver. Only in pancreatic islets gene expression was substantially lower. The levels of the cytoplasmic Cu/Zn SOD and the mitochondrial Mn SOD gene expression were in the range of 30-40% of those in the liver. Glutathione peroxidase gene expression was 15%, and catalase gene expression was not at all detectable in pancreatic islets. These low levels of antioxidant enzyme gene expression may provide an explanation for the extraordinary sensitivity of pancreatic beta cells towards cytotoxic damage by diabetogenic compounds and during the development of human and animal diabetes. PMID- 8720920 TI - A novel HPLC procedure for the analysis of 8-oxoguanine in DNA. AB - The chromatographic quantitation of 8-oxoguanine adducts in DNA is widespread in the literature, although results obtained by HPLC of 8-oxodeoxyguanosine do not always agree with levels determined by GC-MS. To help explain this discrepancy, here we describe a novel procedure for the analysis of 8-oxoguanine adducts in DNA. Although it proved difficult to directly quantitate 8-oxoguanine in the presence of high levels of endogenous guanine using conventional reversed-phase HPLC, a simple preincubation of DNA acid hydrolysates with guanase allowed such analyses. The assay relied on our observation that 8-oxoguanine was not a substrate for guanase, and on sensitive electrochemical detection. The limit of detection for 8-oxoguanine was 5 nM or 250 fmol on column. Using this procedure, the background level of 8-oxoguanine in commercially available calf thymus DNA was 0.4 nmol/mg DNA or 3.2 mol/10(5) mol guanine. PMID- 8720921 TI - N2-methyl-8-oxoguanine: a tRNA urinary metabolite--role of xanthine oxidase. AB - DNA damage produces a series of oxidation products including 8-oxo-2' deoxyguanosine and 8-oxoguanine, whose urinary excretion have been used to estimate in vivo oxidative injury. A monoclonal antibody to 8-oxoguanine was used to measure these adducts in urine taken from ill infants. In the process of this investigation we observed a large chromatographic peak that did not correspond to any of the known 8-oxoguanine adducts. A combination of liquid chromatography with electrochemical detection and gas chromatography/mass spectrometry allowed isolation and identification of the previously undescribed oxidation product, N2 methyl-8-oxoguanine. The excretion of this compound is increased in ill and growing infants but is also found in the urine of adult rats and humans. Experiments with allopurinol, a xanthine oxidase inhibitor, show that in humans, N2-methyl-8-oxoguanine is formed from the tRNA modified base N2-methylguanine. This is in contrast to the nonmethylated bases, which are converted to 8-oxo derivatives as a result of oxidative damage to nucleic acids and do not appear to be substrates for xanthine oxidase. PMID- 8720922 TI - Effect of liver cirrhosis and age on the glutathione concentration in the plasma, erythrocytes, and gastric mucosa of man. AB - GSH and its related enzymes are one of the protective mechanisms vs. the oxidative damage, both in the circulation and in various tissues, including gastric mucosa. Patients with liver cirrhosis frequently suffer from a gastropathy caused by portal hypertension and they present low circulating levels of GSH. Aging processes cause an increase of gastric damage, of lipoperoxidative phenomenons, and a decrease of GSH in animals. The aim of this study was the evaluation, in humans, of the effect of both these factors, age and liver cirrhosis, on the global pool of GSH and on the antioxidant capability of the cells of gastric mucosa. Therefore, we evaluated the effect of liver cirrhosis and age on the circulating levels of GSH, both in the plasma and in the erythrocytes, and the GSH concentration and the activity of the total GSH transferase (GSH-T) in gastric mucosa of healthy subjects and in patients affected by liver cirrhosis. Age, but not liver cirrhosis, induced a significant decrease of GSH and GSH-T activity in gastric mucosa; on the contrary, the plasma levels of GSH decreased in cirrhotics but not in elderly healthy subjects. In the erythrocytes, GSH was affected by both these factors (age and liver cirrhosis). These findings indicate that both in patients with liver disease and in elderly healthy subjects the GSH-related cellular defensive mechanisms are depressed and therefore susceptibility to oxidative damage may increase. PMID- 8720923 TI - Cloning and characterization of a probenazole-inducible gene for an intracellular pathogenesis-related protein in rice. AB - Probenazole (3-allyloxy-1,2-benzisothiazole-1,1-dioxide) induces disease resistance in rice against rice blast fungus. To investigate the molecular mechanism of probenazole-induced resistance, we isolated and characterized a cDNA clone of a probenazole-inducible gene in rice, which encoded a protein designated PBZ1. Sequence analysis revealed that significant homology at the amino acid level exists between the predicted PBZ1 protein and intracellular pathogenesis related (IPR) proteins. Accumulation of PBZ1 mRNA was not induced by wounding, but markedly induced by inoculation with rice blast fungus. In addition, it was induced sooner by inoculation with an incompatible race than that with a compatible race. On the other hand, when the accumulation of the PBZ1 mRNA was examined after treatment with probenazole-related compounds, it was not fully correlated with anti-rice blast activity. However, it was induced after treatment with N-cyano-methyl-2-chloro-isonicotinamide (NCI), which belongs to another group of compounds known to induce disease resistance. Thus, although the accumulation of the PBZ1 mRNA was not fully correlated with anti-rice blast activity, our findings suggest that the PBZ1 gene has an important function during the disease resistance response in rice. PMID- 8720924 TI - Efficient promoter cassettes for enhanced expression of foreign genes in dicotyledonous and monocotyledonous plants. AB - A series of chimeric promoters for higher-level expression of foreign genes in plants was constructed as fusions of a gene for beta-glucuronidase (GUS) with the terminator of a gene for nopaline synthase (nos) or of the cauliflower mosaic virus (CaMV) 35S transcript, and the strength of these promoters was assayed in transient and stable expression systems in tobacco and rice. As parts of these promoters, the CaMV 35S core promoter, three different 5'-upstream sequences of the 35S promoter, the first intron of a gene for phaseolin, and a 5'-untranslated sequence (omega sequence) of tobacco mosaic virus were used in various combinations. In tobacco and rice protoplasts, all three fragments of the 35S promoter (-419 to -90, -390 to -90 and -290 to -90, relative to the site of initiation of transcription), the intron, and the omega sequence effectively enhanced GUS activity. Some chimeric promoters allowed levels of GUS activity that were 20- to 70-fold higher than those obtained with the 35S promoter in pBI221. In tobacco protoplasts, the two longer fragments of the 35S promoter were more effective than the shortest fragment. In rice cells, by contrast, the shortest fragment was as effective as the two longer ones. The terminator of the 35S transcript was more effective than that of the nos gene for gene expression. In transgenic tobacco plants, a representative powerful promoter, as compared to the 35S promoter, allowed 10- and 50-fold higher levels of expression on average and at most, respectively, with no clear qualitative differences in tissue- and organ-specific patterns of expression. When the representative promoter was introduced into tobacco with a gene for luciferase, the autofluorescence of detached leaves after a supply of luciferin to petioles was great and was easily detectable by the naked eye in a dark room. PMID- 8720925 TI - Immunological identification of a putative precursor of the insoluble glycoprotein framework of the Chlamydomonas cell wall. AB - To identify precursors of the insoluble glycoprotein framework of the Chlamydomonas cell wall, a polyclonal antibody was raised against the mixture of polypeptides released from the insoluble wall fraction by chemical deglycosylation. This antibody preferentially cross-reacted with a '150 kDa' salt soluble cell wall glycoprotein. The conclusion that this '150 kDa' glycoprotein is a putative precursor of the insoluble cell wall fraction was corroborated by the results of pulse-chase experiments and by experiments with antibodies raised against the '150 kDa' salt-soluble glycoprotein and against its 100 kDa deglycosylation product, respectively. Whereas the antibody against the '150 kDa' glycoprotein preferentially recognized carbohydrate side chains, the antibody against its 100 kDa deglycosylation product was found to have essentially the same specificity towards glycosylated and deglycosylated cell wall components as the antibody against the deglycosylation products of the insoluble wall fraction. Furthermore, the antibody against the deglycosylated, insoluble wall fraction recognized almost the same set of peptide fragments derived by V8 protease treatment from the '150 kDa' salt-soluble cell wall glycoprotein and its 100 kDa deglycosylation product, respectively, as the antibody against the 100 kDa deglycosylated cell wall polypeptide. PMID- 8720926 TI - Trans-kingdom conjugation between Agrobacterium tumefaciens and Saccharomyces cerevisiae, a bacterium and a yeast. AB - For conjugation between prokaryotic Agrobacterium tumefaciens and eukaryotic Saccharomyces cerevisiae, we constructed two novel conjugative plasmids. A. tumefaciens transmitted the plasmids to S. cerevisiae with the aid of tra genes on a helper plasmid. The transmitted plasmids retained their original structure and function in transconjugant yeasts. The presence of Ti plasmid barely affected the trans-kingdom conjugation. PMID- 8720927 TI - cis-regulatory elements responsible for quantitative regulation of the rice seed storage protein glutelin GluA-3 gene. AB - The 897 bp 5' flanking region of the rice storage protein glutelin gene (GluA-3) was analyzed to define cis-regulatory elements responsible for the seed specific expression by progressive 5' deletions and gain-of-function experiments using a truncated CaMV 35S promoter (-90/+9). These constructs were transcriptionally fused either to the firefly luciferase (LUX) or beta-glucuronidase (GUS) reporter gene and introduced into tobacco. It was shown that a 121 bp sequence between 437 and -317 from the transcriptional start site is important for the quantitative regulation of the GluA-3 gene expression. This region contained at least two functionally different cis-regulatory elements. One acts as a general positive element and the other is involved in a specific negative regulation in non-seed tissues. PMID- 8720928 TI - [Evaluation of pulmonary toxicity of heavy metal compounds]. AB - The present report describes toxicological approaches to evaluate inflammatory potency of heavy metal compounds deposited in the lung. Although inhalation exposure is a well-accepted method to study effects of pulmonary toxicants, it requires expensive facilities and many man-hours to complete experiments. Intratracheal (i.t.) instillation, an alternative and simple method to expose animals to toxicants via airways, has been proved to be useful to investigate the pulmonary clearance of heavy metals. However, acute inhalation exposure caused more severe inflammatory lung injury than i. t. instillation when the same amount of nickel sulfate was administered in the rat. Among several biochemical inflammatory indices such as some enzyme activities and the protein concentration in bronchoalveolar lavage (BAL) fluid, only lactate dehydrogenase (LDH) activity increased linearly with a wide range of doses of heavy metals. The increase in polymorphonuclear leukocytes (PMN) in BAL fluid, a cytological index of inflammatory responses of the lung, is believed to reflect pulmonary toxicity of heavy metals. However, more extensive study is required to elucidate the mechanism of transpulmonary PMN infiltration upon heavy metal insult. In addition to the linearity between LDH activity in BAL fluid and doses of heavy metals, the increase in this cytosolic enzyme activity is associated with cell lysis in the bronchoalveolar milieu and appears to be the best index to evaluate pulmonary inflammatory potency of heavy metals. PMID- 8720929 TI - [Background and prospects of the Community Health Act]. AB - Japan was defeated in World War II and almost all of the nation was demoralized by the destruction and damage to much of the nation. The medical and health care system during and before World War II needed to be reformed radically and fundamentally since almost all medical and health institutes were destroyed. On the other hand, many health personnel came back from overseas after the war. Japanese modern medicine had developed on the basis of German medicine; however, many aspects of American medicine, including public health and democracy, were rapidly introduced following the end of World War II. The American type of health center was established and many laws concerning medical and health care were enacted in 1947-1948. One of them was "The Health Center Law." The National Health Insurance Act was enacted in 1958 and the total population has been covered by health insurance plans since 1961. Many physicians quit the health centers and they have worked as clinicians under the National Health Insurance scheme, because health centers were introduced before adequate education and research existed in the field of public health. On the other hand, the health insurance scheme was in its golden age during the high economic growth period of the 1960s. Japan has succeeded in all forms of modern technology and economy for the past 30 years and is now one of the top nations in the field of medical and health care, such as the numbers of clinics and hospitals and beds, the frequency of consulting with a doctor, length of hospital stay, examinee rates in mass health examinations in the community and workplace and so on. Health conditions have changed drastically from the 1950s to the present. Therefore, health centers do not fit current health needs. For example, mortality from tuberculosis, acute infections diseases and also stomach and uterus cancers and apoplexy have decreased rapidly while mortality from chronic diseases, especially lung, breast and rectal cancers, and myocardial infarction have increased gradually. Changes of life style resulting from rapid economic growth are suspected to be important causes of the change in the prevalence of these diseases. Mass health examination was important and effective as a preventive measure against tuberculosis, especially as a means of early detection and early treatment. However, it is not now effective against chronic diseases. The screening examination has resulted in identifying many patients suspected of being ill. Every examiner must be able to distinguish pathologic findings from physiologic changes of aging. Every patient must, therefore, understand his/her individuality and evaluate the result of his/her efforts to improve life style by receiving a health examination. Accordingly, the aim of health examination has changed from early detection to health support for the examinee. During the decades when life expectancy was less than 50 years of age, it was not necessary for people to plan for retirement. Moreover, there was little burden on younger generations to provide care for the aged people because there were few old people more than 70 years of age and the birth rate was high. Nowadays, elderly people face many years of life after retirement and there are too many aged people in relation to the number of younger persons. As for medical care services, many new medical needs have emerged in recent years, including "quality of life," "palliative medicine in terminal care," "establishment of a primary care system" and "comprehensive care connecting health and medical care with welfare" etc. Improved living standards resulting from economic growth, called the "economic miracle" internationally, have helped to bring about a rapid and wide range of change in daily lifestyle, such as eating habits, working conditions and environment. The Ministry of Health and Welfare has made every effort to revise the laws in relation to health and medical care systems, in order to adjust to recent PMID- 8720930 TI - [Molecular cancer epidemiology--the present status and future possibilities]. AB - Molecular cancer epidemiology is a relatively new strategy for malignancies. This strategy has made it possible to diagnose the predisposition to cancer. An individual is said to have a predisposition to cancer when a tumor-suppressor gene is inactivated in germ-line cells. Mainly the inactivation of the tumor suppressor gene is caused by mutations at the coding region of the gene. However, we clarified that point mutations or hypermethylations of the retinoblastoma tumor-suppressor gene (RB) also cause inactivation of the gene, resulting in retinoblastoma. On the other hand, studies to improve the diagnosed predisposition to cancer have not been performed. We therefore started a basic study for this purpose. As the first example of limiting the predisposition to cancer, the cases with a point mutation at the RB promoter region might be good candidates. In these carriers, only the RB promoter region is inactivated in spite of a lack of abnormalities in the coding region. Therefore, if the RB promoter activity is recovered by drugs, predisposition to retinoblastoma should be limited. As the second example, Li-Fraumeni syndrome in which the p53 gene is hereditarily mutated might be a good candidate. Recently p53 has been reported to stimulate the WAF1 gene, and the WAF1 protein to inhibit cdk activity, which inactivates the RB gene product by phosphorylation. In addition, we found that p53 up-regulates the promoter activity of the RB gene. These findings suggest that p53 directly or indirectly activates RB at the transcriptional or post transcriptional level. Therefore, reactivation of the WAF1 or RB gene by certain drugs might compensate for the loss of function of p53 in Li-Fraumeni syndrome. We then suggest that it might be possible to prevent cancer by enhancing some intact target genes of the genetically inactivated tumor-suppressor gene. We term this new strategy "gene-regulating chemoprevention." To test this hypothesis it is important to clarify the structure of the RB promoter. In summary we found that RBF-1 and ATF sites are the core promoter regions, that the neighboring E2F site is a silencer site, and that E4TF1 preferentially binds to the RBF-1 site. We then speculate that drugs interfering with the binding of the E2F complex might become good candidates enhancing RB promoter activity. To find drugs regulating the promoters of these genes, it is reasonable to try G1 arresting drugs, because WAF1 and RB are thought to arrest cells at the G1 phase. Actually we found several drugs causing G1 arrest. They are several flavonoids which we ingest daily from vegetables and fruits, or prostaglandin D2 and its metabolite. In summary, we propose that "gene-regulating chemoprevention" will be a useful method for molecular cancer epidemiology. PMID- 8720931 TI - [Effects of long-term physical training on body composition, cardiovascular function, and serum lipids in mildly obese middle-aged subjects]. AB - To investigate the effects of long-term physical exercise on humans, body composition, cardiovascular function and serum lipids were measured for a training period of 20 months in 8 men and 8 women, aged 36-67 years who were mildly obese. The subjects participated in physical training for 90 minutes about twice a week for 20 months. The intensity of training was at most 60% VO2max for each subject as calculated by karvonen's formula. The results are summarized as follows: (1) Body weight and estimated body fat weight decreased from baseline after the training in both sexes. The percentage decreases in body weight and estimated body fat weight in men were about threefold those in women. (2) Following the training, heart rate, blood pressure and rate pressure product (RPP) at rest had significantly decreased from initial levels in both sexes. (3) Following the training, the physical work capacity at a heart rate of 150bpm significantly increased, and systolic blood pressure at the heart rate of 150bpm significantly decreased from baseline levels in both sexes. (4) Following the training, the serum HDL-cholesterol and HDL-/total-cholesterol ratio significantly increased from baseline, and serum triglycerides and total cholesterol also significantly decreased from baseline levels in both sexes. These results suggest that long-term physical training with moderate intensity improves body composition, cardiovascular function and serum lipids in mildly obese middle-aged men and women, and reduces the risks for coronary heart disease. PMID- 8720932 TI - [Relation among lifestyle, social network, blood pressure and serum lipids in the elderly]. AB - The relationships among seven lifestyle factors including stress, which is focused on as a risk factor for cardiovascular disease recently, social network, blood pressure and serum lipids in the elderly were examined in a cross sectional study of 85 males and 65 females aged 60 years or over in the city of Sayama, Saitama Prefecture. The seven lifestyle factors were as follows: (1) amount of alcohol consumption, (2) cigarette smoking, (3) Physical exercise, (4) sleeping time per night, (5) nutritional balance, (6) body weight, and (7) stress. As social network factors, marital status and coping factor for stress and/or anxiety were introduced. Stressful life events strongly associated with stress status were also discussed. In this study, it was clear that age, alcohol consumption, stress status and stressful life events caused higher blood pressure. On the other hand, the social network was associated with lifestyle status and a controlled blood pressure level. Obesity and physical exercise were significantly associated with serum lipids. There results suggest that it is very important for the maintenance and promotion of health and risk reduction of disease among the elderly to control socio-psychological factors such as stress and stressful life event, and that a social network helps maintain a healthy lifestyle. PMID- 8720933 TI - [Accidental injuries among elementary school children and some of their environmental factors]. AB - I investigated 4960 accidental injuries which occurred among elementary school children between April 1991 and March 1992, using reports of school accidents submitted to the Shiga branch of the National Studium and School Health Center of Japan. Data on six school environmental factors (i.e. the areas of school land, schoolhouse, gymnasium and all classrooms, the number of children per class, and the total number of children in each school) for 223 elementary schools were gathered. From these data, the relationship between the incidence rates of injuries and the above school environmental factors were analyzed. The overall average incidence rate for all injuries combined was 5.43 cases per 100 children. For all injuries and fractures which occurred at smaller schools, the incidence rates tended to increase with the areas of school land, schoolhouse, gymnasium and all classrooms, and to decrease as the number of children per class and the number of children in the school increased. These results may have been biased by the fact that only injuries treated at a cost of at least 3000 yen were reported. In the process of stepwise multiple regression analysis between the injury-rates and environmental factors, a few variables relating to types of injuries were included as statistically significant factors in the well-fitted equations, but the coefficients of determination (squared multiple correlation coefficients) in the equations were small. PMID- 8720935 TI - [A population survey on bone mineral density in a fishing village in Wakayama prefecture. (Part 1) Distribution of bone mineral density by sex and age based on a representative sample of the community]. AB - To establish reference values for bone mineral density (BMD), a population survey was carried out in a fishing community in Wakayama Prefecture. The BMD measurements of the lumbar spine (L2-L4) and proximal femur (femoral neck, trochanter and Ward's triangle area) were performed by sex and age, using dual energy X-ray absorptiometry. Four hundred subjects, aged 40 to 79 years, were recruited randomly to give 50 persons in each of eight age-sex strata from a list of 2,261 residents (1,028 men and 1,233 women) living in a fishing village. The validity of sampling methods was assessed using a questionnaire about lifestyle factors, which was used for all residents aged from 40 to 79 years in the baseline survey. There were no significant differences between the subjects for BMD measurements and all the residents of the community in the frequencies of past history of diseases, healthy habits, food intakes, and rates of smoking, alcohol drinking, coffee and green tea consumption. These findings suggested that the present study population could be considered representative of samples obtained from the entire population in the fishing community. The results of BMD measurements in these subjects, who were selected at random, showed that except for a slightly high BMD of the lumbar spine in men in their 60's, the mean BMD of the lumbar spine and the three proximal segments of the femur decreased with increasing age in both sexes. PMID- 8720934 TI - [The changes of complement activities in sera of mice after subcutaneous administration of beryllium chloride]. AB - We studied changes of the complement pathway activities and the content of C3 in sera of mice, administered BeCl2 (containing 5 micrograms of Be per mouse) or CuCl2 (containing 5 micrograms of Cu per mouse) by a single subcutaneous injection. The value of the classical complement pathway activity (CH50) of the Be group 3 days after administration was significantly higher than that of the control group (P < 0.001). It was significantly lower than in the control group after 7 days (P < 0.001). On the other hand, the CH50 value of the Cu group 3 hr after administration tended to increase, however, it was significantly lower than in the control group after 7 days (P < 0.01). The change of the alternative complement pathway activity (ACH50) value of the Be group was similar to the change of the CH50 value of the group. The ACH50 value of the Cu group 3 days after administration tended to increase but it was the same as the ACH50 value of the control group after 7 days. The C3 contents of both the Be and Cu groups 3 days after administration were significantly higher than in the control group (P < 0.001). The aspartate aminotransferase (AST) activity of the Be group 7 days after administration was significantly higher than that of the control group (P < 0.01). By contrast, AST activity of the Cu group 3 hr after administration was significantly higher than in the control group (P < 0.05). The value of the alanine aminotransferase (ALT) activity of the Be group was low (P < 0.01), but that of the Cu group was high (P < 0.05), 3 hr after administration. These values of both groups after 7 days, however, were significantly higher than in the control group (P < 0.05). The AST/ALT ratio in mice was very high at 3 hr, and it remained high by 7 days after Be injection. On the other hand, the ratio of the Cu group was almost constant for 7 days after Cu injection. Thus, these values changed with relative expedition after Be injection. Therefore, we confirmed that measurements of complement activities and the content of C3 were valuable indices for assaying acute effects of Be on mice. PMID- 8720936 TI - [The effects of UVA irradiation on human neutrophil function]. AB - We investigated the in vitro effects of ultraviolet A (UVA) irradiation on human neutrophil function. For measurements of reactive oxygen species (ROS) and serum opsonic activity, chemiluminescence was used. Chemotactic activity was determined by the Boyden technique. In this study, for detection of the effects of UVA irradiation on human neutrophil ROS production, we used luminol-dependent and lucigenin-dependent chemiluminescence, in addition to two kinds of action stimuli, opsonized zymosan and phorbol myristate acetate (PMA). We found that UVA irradiation at low doses of UVA (0.2-0.4J/cm2) did not affect the generation of OC1-, and it was suggested that high doses of UVA (0.6-1.0J/cm2) suppressed OC1- production by inactivating the signalling cascade which induces ROS on the membrane surface of neutrophils. No significant difference was observed in the chemotactic activity. The serum opsonic activity was increased by UVA irradiation. These results suggest that low doses of UVA do not impair neutrophil phagocytic activity, and that higher doses of UVA suppress the ROS generating capacity of neutrophils. However, to compensate for ths suppression, serum opsonic activity was induced, so it seemed that the neutrophil-related immune system could be retained. PMID- 8720937 TI - Commitment to division in ciliate cell cycles. AB - Near the end of the cell cycle, ciliates commit irreversibly to cell division. The point of commitment occurs at the time of oral polykinetid assembly and micronuclear anaphase. The commitment is a checkpoint which requisites a threshold cell mass/DNA ratio and stomatogenesis. It is also a physiological transition point, involving cdk protein kinases similar to those of other eukaryotes. Both P34 kD and P36 kD kinases, similar to the S. pombe cdc2 kinases, have been described to have activity as monomers. Subsequent to commitment to division, dramatic cytoskeletal modifications occur for separation of organelles, cortex morphogenesis and cytokinesis. Numerous mutants affecting cytoskeletal function associated with the division process have been obtained in several species. Of these, only the cc1 mutant in Paramecium affects cell cycle progression prior to commitment to division. The material reviewed is used to speculate about the mechanisms of regulation of pre-fission morphogenesis and cell division related processes in ciliates. PMID- 8720938 TI - Symposium introductory remarks: "protistan molecular ecology and systematics". PMID- 8720939 TI - Ribosomal RNA-based oligonucleotide probes to identify marine green ultraphytoplankton. AB - Oligonucleotide probes based on small subunit 18S rRNA variable region sequences, were designed and used to discriminate groups of marine green ultraphytoplankton of similar size and morphology. Hybridization of probes was visualized using a combination of biotinylated oligonucleotides and fluorescein labeled avidin. Increased fluorescent signal was generated by building a complex of biotinylated probe, fluorescein labeled avidin and biotinylated anti-avidin. Cells of one phylogenetic group were easily differentiated from morphologically similar cells of a different phylogenetic group. Design of specific probes allowed identification of Mantoniella squamata, Micromonas pusilla, Pseudoscourfieldia marina and the micromonadophyte clone CCMP1194. This technique should be readily adaptable to open ocean samples and should greatly facilitate the description and quantification of ultraphytoplankton in the open ocean. PMID- 8720940 TI - Molecular identification of nanoplanktonic protists based on small subunit ribosomal RNA gene sequences for ecological studies. AB - Nanoplanktonic protists are comprised of a diverse assemblage of species which are responsible for a variety of trophic processes in marine and freshwater ecosystems. Current methods for identifying small protists by electron microscopy do not readily permit both identification and enumeration of nanoplanktonic protists in field samples. Thus, one major goal in the application of molecular approaches in protistan ecology has been the detection and quantification of individual species in natural water samples. Sequences of small subunit ribosomal RNA (SSU rRNA) genes have proven to be useful towards achieving this goal. Comparison of sequences from clone libraries of protistan SSU rRNA genes amplified from natural assemblages of protists by the polymerase chain reaction (PCR) can be used to examine protistan diversity. Furthermore, oligonucleotide probes complementary to short sequence regions unique to species of small protists can be designed by comparative analysis of rRNA gene sequences. These probes may be used to either detect the RNA of particular species of protists in total nucleic acid extracts immobilized on membranes, or the presence of target species in water samples via in situ hybridization of whole cells. Oligonucleotide probes may also serve as primers for the selective amplification of target sequences from total population DNA by PCR. Thus, molecular sequence information is becoming increasingly useful for identifying and enumerating protists, and for studying their spatial and temporal distribution in nature. Knowledge of protistan species composition, abundance and variability in an environment can ultimately be used to relate community structure to various aspects of community function and biogeochemical activity. PMID- 8720941 TI - Ca(2+)-dependence of conoid extrusion in Toxoplasma gondii tachyzoites. AB - The role of Ca2+ in conoid extrusion was investigated in isolated Toxoplasma gondii tachyzoites by treatment with Ca(2+)-ionophores, Ca(2+)-chelating agents and an inhibitor of the Ca(2+)-ATPase at the endoplasmic reticulum. The results were evaluated by light phase-contrast microscopy and electron microscopy. Ionomycin (0.5-1 microM) caused an immediate and sustained extrusion of the conoid in up to 80% of the tachyzoites, depending on the concentrations of ionophore and Ca2+ in the medium. However, over 50% of the tachyzoites extruded the conoid when treated with ionomycin in Ca(2+)-free saline complemented with EGTA. The effect of ionomycin was reversible and could be induced a second time in about half of the responsive population. Similar results were obtained with A23187. Conoid extrusion induced by ionomycin in Ca(2+)-free medium was almost completely abolished when the tachyzoites were previously loaded with a permeable compound known to chelate intracellular Ca2+ (BAPTA/AM; 25 microM). On the other hand, exposure of tachyzoites to the Ca(2+)-ATPase inhibitor thapsigargin (0.5-1 microM) produced significant extrusion of the conoid. Tachyzoites loaded with BAPTA/AM as well as those treated with ionomycin, i.e. with conoids paralyzed in opposite positions, had a diminished capacity to invade cultured epithelial cells. A substantial reduction in the response to stimulation by ionomycin was found also in parasites treated with cytochalasin-D, a drug that depolymerizes actin-filaments. The results suggest that Ca(2+)-release from internal stores may act as a key signal to activate a mechanism of conoid extrusion probably mediated, at least in part, by actin-filaments. PMID- 8720942 TI - DNA polymorphism revealed by arbitrary primers polymerase chain reaction among Blastocystis strains isolated from humans, a chicken, and a reptile. AB - DNA polymorphisms of different strains of Blastocystis isolated from humans, a chicken, and a reptile were examined by an arbitrary primer PCR method. Two strains of Blastocystis hominis isolated from humans in the USA and Japan yielded nearly identical PCR products. However, one strain of B. hominis (isolated from a human in Singapore) yielded quite different PCR products. Blastocystis sp. isolated from a chicken yielded PCR products similar to those of the former two strains, while Blastocystis lapemi, isolated from a reptile, shared no bands with any of the other isolates. These results indicate the possibility that our isolate from the chicken is a zoonotic strain, and that there is intraspecific variation of Blastocystis hominis. PMID- 8720943 TI - Ubiquity of cysteine- and metalloproteinase activities in a wide range of trypanosomatids. AB - We have analysed the proteinase profiles of 11 species from 7 different genera of trypanosomatids by in situ detection of enzyme activities on SDS-PAGE gels containing co-polymerized gelatin as substrate, and the use of specific proteinase inhibitors. Our survey indicates that while cysteine- and metalloproteinases are distributed ubiquitously among trypanosomatids, there are marked differences between the enzyme profiles from the monogenetic (Crithidia, Herpetomonas, Leptomonas) and digenetic (Trypanosoma, Endotrypanum, Phytomonas, Leishmania) species. The detected metalloproteinase activities, ranging in size from 50-100 kDa, partitioned into the detergent-phase after Triton X-114 extraction, while most of cysteine proteinases, of three distinct molecular mass ranges (30-50 kDa, 80-100 kDa and 116-205 kDa), partitioned into the aqueous phase. Thus, within this group of organisms, the metalloproteinase activities seem to be predominantly membrane-associated proteins. We also show that the plant parasites of the genus Phytomonas exhibit a distinctive cysteine proteinase profile that might be exploited further as a criterion for taxonomy of the genus. PMID- 8720944 TI - Oxidation of alanine, acetate, glutamate, and succinate by digitonin permeabilized Leishmania major promastigotes. AB - Leishmania major promastigotes were treated with digitonin and the rates at which [1-14C]acetate, [1,4-14C]succinate, [1-14C]glutamate, and [U-14C]alanine are oxidized were measured in the presence of suitable cofactors. Acetate was oxidized at the lowest rate of the four substrates examined, even in the presence of added NAD, CoA, ADP and acetyl-CoA synthase. Its rate of oxidation was negligible if the permeabilized cells were washed before the cofactors were added, indicating the requirement for an as yet unknown factor. Succinate was oxidized at a rate much higher than the very slow rate at which it is oxidized by intact cells. Its rate of oxidation was strongly inhibited by antimycin A, but that of glutamate was scarcely affected. Fumarate inhibited the rate of oxidation of acetate, glutamate, and succinate, but increased that of alanine. Ca++ inhibited the rates of oxidation of alanine and succinate, but not of acetate or glutamate. Increasing the osmolality by addition of mannitol partially inhibited the rate of oxidation of alanine but had little effect on that of glutamate. These results show that appreciable transaminase activity remains in the permeabilized cells and support earlier data indicating the presence of a branched NAD-to-cytochrome oxidase system. These results also provide preliminary information on the sensitivity of the two branches to Ca++, hyperosmolality, and Krebs cycle intermediates. PMID- 8720945 TI - Chemosensory responses of Acanthamoeba castellanii: visual analysis of random movement and responses to chemical signals. AB - A visual assay slide chamber was used in conjunction with time-lapse videomicroscopy to analyze chemotactic behavior of axenically grown Acanthamoeba castellanii. Data were collected and analyzed as vector scatter diagrams and cell tracks. Amebas responded to a variety of bacterial products or potential bacterial products by moving actively toward the attractant. Responses to the chemotactic peptide formyl-methionyl-leucyl-phenylalanine (fMLP), lipopolysaccharide, and lipid A were statistically significant (P < or = 0.03), as was the response to fMLP benzylamide (P < or = 0.05). Significant responses to cyclic AMP, lipoteichoic acid, and N-acetyl glucosamine were also found. Chemotactic peptide antagonists, mannose, mannosylated bovine serum albumin, and N-acetyl muramic acid all yielded nonsignificant responses (P > 0.05). There was no single optimal concentration for response to any of the attractants tested, and amebas responded equally over the range of concentrations tested. Pretreatment of amebas with chemotactic peptides, bacterial products, and bacteria reduced the directional response to attractants. Amebas that had been grown in the presence of bacteria appeared more responsive to chemotactic peptides. Treatment of amebas with trypsin reduced the response of cells to chemotactic peptides, though sensitivity was restored within a couple of hours. This suggests the ameba membrane may have receptors, sensitive to these bacterial substances, which are different from the mannose receptors involved in binding bacteria to the membrane during phagocytosis. The rate of movement was relatively constant (ca. 0.40 microns/s), indicating that the locomotor response to these signals is a taxis, or possibly a klinokinesis, but not an orthokinesis. Studies of the population diffusion rate in the absence of signals indicate that the basic population motility follows the pattern of a Levy walk, rather than the more familiar Gaussian diffusion. This suggests that the usual mathematical models of ameboid dispersion may need to be modified. PMID- 8720946 TI - Phylogeny of the large extrachromosomal DNA of organisms in the phylum Apicomplexa. PMID- 8720948 TI - [Clinical case. Retention cyst of Bartholin's glands]. PMID- 8720947 TI - [Antifreeze glycoproteins in fishes: structure, mode of action and possible applications]. AB - Two types of antifreezes have been isolated from polar and northern temperate fishes so far. They are either glycopeptides or peptides. Whereas these proteins have only a very small effect on the melting temperature of ice, the temperature of these fish can fall to nearly 1 degree below the melting point before ice crystals grow. This phenomenon is called thermal hysteresis, in contrast to the normal colligative effect of solutes. All Antarctic notothenioids (perches) investigated so far have the typical antifreeze glycoproteins (AFGP) with the tripeptide Ala-Ala-Thr and the disaccharide Gal-GalNAc. In the Antarctic silverfish Pleuragramma antarcticum there could be found a novel GlcNAc containing antifreeze glycoprotein, the PAGP. The antifreezes not only lower the freezing temperature, but they also retard recrystallization on frozen storage. Antifreeze proteins thus could be useful for biotechnology and cryomedicine in the future. Since some are now synthesized chemically or by genetic engineering, they no longer have to be isolated from fish blood. PMID- 8720949 TI - [Electron microscopic study of cells of the lactating bovine mammary gland]. AB - In this study all cells of the normal bovine mammary gland are described by means of electron microscopic investigations. The ultrastructural features and differences of the alveolar cells of the mammary ducts, the myoepithelial cells, the lymphocytes, the fibroblasts, the fibrocytes, the mast cells, and the macrophages are emphasized. The paper aims to identify the various cells in the milk. PMID- 8720950 TI - [Nephrectomy for chronic, unilateral suppurative pyleonephritis in cattle]. AB - This report presents the case history of a five-year-old Eringer cow suffering from chronic hematuria. Results of clinical examination, ultrasonography of the kidneys, endoscopy of the bladder, and cytologic and bacteriologic analysis of the urine revealed unilateral pyelonephritis of the right kidney caused by Corynebacterium renale. Antimicrobial treatment with procaine penicillin was not successful. Therefore, nephrectomy of the affected kidney was performed through a right flanc approach in the standing animal. Follow-up examination ten months after surgery revealed normal general condition. Etiopathogenesis and diagnostic and therapeutic procedures of pyelonephritis in cattle are discussed. PMID- 8720952 TI - [Mammary gland hypoplasia and aplasia (inverted nipples) in female and male swine. 1. Clinical-morphological investigations on the occurrence and development of inverted nipples in female and male swine of various age and production groups]. AB - The development of the shape of teats in 30 female piglets, of which at least one of the parents showed inverted teats, was systematically observed during three to four months. Similar investigations could be carried out in sows with inverted teats from day 60 of pregnancy until the end of lactation. There is no evidence that inflammations or traumatas are causes for teat hypoplasias, and there is no correlation between "teat necroses" of piglets and the incomplete teat development. The wide distribution of teat abnormities is shown by investigations of slaughter pigs: 20% of fattening pigs, 17% of boars and 6% of sows were affected by inverted teats. 75% of the inverted teats were seen periumbilical. The morphology of hypo- and aplastic teats in boars is described. With regard to the selection of affected pigs, a first investigation is recommended in the 12th week of life and a second one at the age of five month is recommended. Particular attention has to be paid to boars for mating and artifical insemination in order to exclude affected boars as early as possible from breeding. PMID- 8720951 TI - [Reference values of electrodiagnostic and laboratory studies in young Wallis Schwarzhals goats]. AB - Laboratory testing in 10 "Walliser Schwarzhals"-goats included red and white blood cell count, electrolytes, liver enzymes, trace elements and urinalysis, protein determination and cell count of the cerebrospinal fluid (CSF) was also performed. Electrodiagnostic testing included electromyography (EMG) of the head and spinal muscles, motor nerve conduction velocity (MNCV) of the right and left peroneal nerve, brainstem auditory evoked potentials (BAEP) and electroencephalography (EEG). MNCV of the peroneal nerve was 72 +/- 9.24 m/s. Latency and amplitude of the BAEP for P1 was 2.15 +/- 0.08 ms respectively 2.23 +/- 0.7 microV and for P5 5.09 +/- 0.15 ms respectively 0.68 +/- 0.24 microV. P2 (3.2 +/- 0.14 ms/1.37 +/- 0.32 microV) and P3 (3.9 +/- 0.13 ms/0.84 +/- 0.14 microV) were equally distributed between P1 and P5. Whereas the results of EMG and BAEP were similar to those of other species, the electroencephalographic findings consisted of three different patterns depending on duration and depth of isofluran anesthesia. Pattern I had a rhythmic high amplitude (80 microV) and low frequency (1-2 Hz) pattern with aperiodic intervals of relative electric silence. Pattern II was characterized by increased duration of HALF (150 microV), associated with spindles of low amplitude (20-30 microV) and high frequency (10 Hz). Pattern III had generalized delta-activity of high amplitude (70-200 microV). All the results are discussed briefly. PMID- 8720953 TI - [Polyostotic hyperostosis in a plum-headed parakeet (Psittacula cyanocephala L., 1766]. AB - With emphasis on a case report, the clinical and pathological findings commonly associated with polyostotic hyperostosis as well as its etiology are discussed, and the available literature is reviewed. The diagnostic specifics of the disease are detailed, and a survey of possible differential diagnoses is given. PMID- 8720954 TI - [Luxation of the shoulder joint in a horse. A case report]. AB - Etiology, diagnosis and therapy of the luxation of the shoulder joint in the horse are described. The traumatic luxation is diagnosed by physical examination and radiography. Without complicating changings (like fractures) and in case of early correction the prognosis is good. PMID- 8720955 TI - [Congenital bile duct cysts and liver fibrosis in a foal]. AB - A three-week-old foal was submitted to the clinic because of a minor traumatic injury at the lower jaw. At admission the foal exhibited diarrhea, a distended abdomen and reduced general condition. These findings could not be associated with the injury. On abdominal radiography decreased abdominal detail and a dorsocaudal displacement of the intestine was present. Ultrasonographically multiple fluid-filled cystic structures of several centimeters in diameter were identified. These cystic structures appeared to be associated with the liver. At laparotomy and at necropsy the liver was markedly enlarged and firm and had large, thin-walled, with a bile-like fluid filled cysts, 4-20 centimeters in diameter. Histological lesions were characterized by proliferation of small bile ducts and of interlobular connective tissue as well as focal subacute cholangitis. Gross and histological findings were considered to be consistent with congenital polycystic liver disease and fibrosis with ascending cholangitis. PMID- 8720956 TI - [Spectrum of species and incidence of endoparasites in foals and their mother mares from breeding farms with and without anthelmintic prophylaxis in upper Bavaria]. AB - In this epidemiological study 37 foals and mares were included, which have been coproscopically examined in regular intervals over a period lasting up to 13 months. The animals derived from nine breeding farms. In three of these farms no anthelmintics were used before this investigation, whereas in six farms at least three years before regular prophylactic treatments with anthelmintics have been carried out. This investigation revealed no differences in the species spectrum of endoparasites as well as in the incidence of infections as regards to the breeding farms with and without anthelmintic prophylaxis, however, significant differences between foals and mares. At least nine species were established in foals, Strongyloides westeri in 54.1% of the animals, Parascaris equorum in 64.9%, small strongyles, whose third stage larvae showed either eight or 12 intestinal cells, in 83.8%, Anoplocephala perfoliata in 16.2%, Eimeria leuckarti in 64.9%, Giardia in 5.4%, Cryptosporidium parvum in 2.7% and Gasterophilus intestinalis in 21.6%. Parasitic infections with two or more species were found in 83.8% of the foals. The species spectrum of endoparasites in mares from breeding farms without anthelmintic prophylaxis consisted of at least three species of small strongyles, whose third stage larvae were characterized by eight, 12 or 16 intestinal cells. In mares from breeding farms practising regular anthelmintic prophylaxis, however, at least seven species were ascertained, the same small strongyles, P. equorum, A. perfoliata, E. leuckarti and G. intestinalis. PMID- 8720957 TI - [Prolapse of the eyeball in small animals: a retrospective study of 36 cases]. AB - Proptosis of the globe is an ophthalmic emergency, which requires rapid assessment of the situation and immediate medical and surgical therapy. The relevant prognostic signs as well as the medical and surgical management are discussed. A retrospective study of 36 proptosed globes at the Zurich Veterinary School revealed that young animals with a mean age of 5.2 years (dogs) and 4.7 years (cats) are at risk. In the dog both sexes are equally at risk, while tomcats have a higher incidence than females. In ten cases immediate enucleation of the proptosed globe was necessary. The remaining eyes could be replaced. The majority of the repositioned globes were subsequently blind, the cosmetic result, however, was satisfactory according to most owners. Ophthalmic follow-up examinations revealed a large number of long-term damages in repositioned eyes in dogs and cats concerning anterior and posterior segment. PMID- 8720958 TI - [Experiences with propofol as an injection narcotic for radiotherapy of cats and dogs]. AB - In a clinical study propofol has been used intravenously for multiple anesthesias. The results show that repeated anesthesias are possible, even in geriatric patients; patients need no premedication; maintenance of desired anesthetic depth is easily achievable by titrating the propofol dose; quality of anesthesia has been entirely satisfactory for our purposes; recovery is rapid, free from agitation and complete. However, propofol is rather expensive and any unused propofol must be discarded immediately after the anesthesia due to the danger of microbial contamination. PMID- 8720959 TI - [Percutaneous cystoscopy in dogs and cats]. AB - Cystoscopy enables a reliable diagnosis of urinary bladder diseases. The use of transurethral or perineal endoscopic technique is limited by anatomical features. Therefore it was examined, if the prepubic percutaneous access shows advantages referring to diagnosis and treatment. Using this technique three dogs and two cats were examined. The endoscopic instrumentarium and the technical procedure are described. PMID- 8720960 TI - [Biological role of movement in the temporomandibular joint in sheepdogs and house cats]. AB - The temporomandibular joints of the dog and the cat differ in morphology and movement. Characteristic differences in the teeth and the movement of the mandibula result in species-specific mastication: The dog grinds and crunches food. Moving the molars vertically, the cat cuts food to pieces. It has been postulated that the cat masticates more thoroughly than the dog. This is not true. The unilateral mincing of the cat is due to the extremely secodont teeth without possibility to grind. PMID- 8720961 TI - [Infectious burdens of reproduction of female dogs]. AB - The results of gynecological investigations in 142 bitches were evaluated and the complexity of infectious affections is discussed. High proportions of infectious cases were found in cases of limitation of fertility (67.5%), in vaginal discharge in the estrus (60.8%), in cases of mastitis/pseudopregnancy (61.5%) and in mortality of newborn puppies. St. aureus and E. coli were often isolated. There is a high etiological correlation in epidemiology of diseases of the reproductive tract in the single bitch and especially in kennel bitches. PMID- 8720962 TI - [Clinical investigations of an i.m. combination anesthesia with fentanylclimazolam/xylazine and postoperative i.v. antagonism with naloxone/sarmazenil/yohimbine in guinea pigs]. AB - Aim of the study was the clinical use of a completely antagonisable anaesthesia in guinea pigs. The dosage based on experimental studies. Fentanyl, climazolam and xylazine (0.05/2.0/2.0 mg/kg) were injected i.m. for combination anaesthesia and naloxone, sarmazenil and yohimbine (0.03/0.3/2.0 mg/kg) i.v. were used for antagonisation. 18 guinea pigs (17 female, one male) with an average body weight of 1177 g aged between eight months and five years were anaesthetised for various surgical procedures. The following clinical parameters were measured: body temperature, muscle relaxation, analgesia, hyperacusia, righting-, interdigital- and palpebral reflex. After antagonisation the return of righting reflex, the appearance of muscle trembling, the first walking without ataxia and the first food uptake were observed. The investigations showed that this kind of anaesthesia is suitable for surgical procedures in the guinea pig and is very safe because of the complete antagonisation of the depressing respiratory and circulatory effects of anaesthesia. PMID- 8720963 TI - [Immune response of fishes with respect to phylogenetic aspects]. AB - The immune response progressed by steps in the phylogeny of animals. The non specific immune response has been improved by specific immune mechanisms during the early evolution of the organisms. Typical B- and T-cells as well as true antibodies originated first in fishes during the phylogenesis. Fishes possess immune mechanisms which appear to be homologous to those of higher vertebrates. PMID- 8720964 TI - [Characterization of chick embryo fibroblast culture--overview of its effectiveness as a hybridoma cell]. AB - The use of culture supernatant (added in a final concentration of 25% v/v) from chicken embryonal fibroblasts (CEF) for stimulation of established hybridoma cell lines has been shown. The proliferative effect on hybridoma cells from different origins was heatproof and was possible with a reduced serum part (FCS) in the culture medium. After a chromatografic separation the supernatant sections were tested and the positive effect was much lower in comparison to the total conditioned media. PMID- 8720965 TI - [Reference: article of Mr. Prof. Dr. W. Kraft: Clinical veterinary medicine in Germany--quo vadis? (Tierarztliche Praxis 1995; 23:224-51)]. PMID- 8720966 TI - [Discussion group: for caregivers confronted with death in the hospital environment]. AB - Caregivers facing death and supporting dying patients don't have often the possibility to express their suffering inherent in their practice. Repression of affects can be harmful increased by all the "unsaid" and leads to splitting defence mechanisms among them. Caregivers will verbalize this suffering if a specific space and time for talking is provided. Paradoxically the medical team found it unnecessary and equated to a "one more meeting". A talking group is considered as a tool for the caregivers and will adapt itself to the institutional functioning. It has not to be denatured from its real function. It has to avoid pitfalls such as paying back conflicts. This group will survive the institutional assaults if some specific requirements are respected. Thus, the group will achieve, or at least facilitate better relationships between caregivers and patients. PMID- 8720967 TI - [Clinical experience with a novel surgical technique: laparoscopy-assisted vaginal hysterectomy]. AB - A new surgical technique, the laparoscopically assisted vaginal hysterectomy (LAVH) is evaluated through the clinical course of a group of patients operated for hysterectomy in the years 1993-1995. Although the groups of patients being operated for LAVH, vaginal or abdominal hysterectomy have not been randomized, we conclude that the laparoscopic assistance allows the vaginal approach of hysterectomies that should have been otherwise operated by laparotomy. Medical and financial advantages of this technique are discussed. PMID- 8720968 TI - [Response to 6 questions apropos transient cerebral ischemia]. AB - The author answers to six questions relating to the cerebral transitory ischemic attacks, namely definition, significance, frequency, diagnosis, causes and prevention. Within this context, the actualization of antiagregation, anticoagulation and endarterectomy is presented. PMID- 8720969 TI - [We had an unlimited budget and we have exceeded it...]. AB - The extraordinary costs involved in medical innovation leads, inevitably, to financial restrictions. Doctors need to involve themselves in the decision making process, difficult as this may be. Otherwise, health administrators will assume the full responsibility for decision making, as has become the case in the United States. Doctors must be particularly wary that financial restrictions impinge on neither medical progress--through subtle cut-backs--nor on the fundamental ethical principals--such as a decision to reduce the intensity of care in patients with complications. PMID- 8720970 TI - [Postpartum psychiatry]. AB - The four important syndromes that constitute postnatal psychiatry are chronologically: 1) Post-Traumatic Stress Disorder (PTSD), 2) Baby-blues, 3) Puerperal psychosis and 4) Postnatal depression. The PTSD (prevalence 1%) occurs from 24 to 48 hours after a difficult childbirth. Anxiety predominates in association with nightmares and recurrent images of the experience. Baby-blues (prevalence 80%) is a feeling of sadness that doesn't need to be treated. It occurs around the third day of post-partum. Its intensity is a predictive factor of postnatal depression. Puerperal psychosis (prevalence 0.2%) is a disturbance of psychotic or manic proportion, mainly in women with a psychiatric previous history. It occurs between the first and the third week of the post-partum period. Postnatal depression (prevalence 20%) is a major depressive state commencing four to six weeks after delivery. It is rarely diagnosed early and child abuse is one of its consequences. Prevention--the important point of the treatment--is most of the time neglected. PMID- 8720971 TI - [Exposure to ionizing radiation: radiobiological effects and pathogenesis. 1]. AB - The present paper describes the radiobiological effects induced by an exposure to ionizing radiation and their pathogenesis. The different skin reactions are described in detail because of their importance and frequency. Thus the acute skin lesions after high doses and the late effects resulting, either from high doses, or from accumulation of chronic irradiation, are studied. The main early syndromes are then characterized: neurological, gastro-intestinal, bone-marrow and prodromic. As far as the complex problem of radiocarcinogenesis is concerned, the main results derived from studies by international organizations such as the ICRP and the UNSCEAR are reported: risk coefficient of 5% per gray, for lethal radio-induced cancer, after total body irradiation, at low dose of low-LET radiation. The effects of irradiation in utero are then considered: risk of malformation after irradiation during the two first months of pregnancy and risk of mental retardation after irradiation during the third and the fourth months. Finally, the genetic risk is presented as being equal to one fourth of the risk of carcinogenesis at low-doses. The effects of irradiation on the gonads are also described. PMID- 8720972 TI - [Nuclear Magnetic Resonance Imaging in the staging of adenocarcinoma of the uterine cervix]. AB - Clinical staging is often inaccurate in the evaluation of local extension of cervical carcinoma. In the present study the sensivity of MR imaging in predicting extracervical tumor invasion on the basis of the detection of a thinning of the univolved cervical stromal ring (3 mm or less) was compared to the sensivity achieved by direct visualization of a complete stomal interruption. Fifteen consecutive patients with adenocarcinoma of the uterine cervix (FIGO stage Ib-IIa), were examined with axial and sagittal weighed MR sequences. Pathologic proof after hysterectomy was obtained in all subjects. The sensitivity in predicting extracervical involvement on the basis of the MR Imaging visualization of a thinned stroma was 83%, whereas the sensitivity obtained by detection of a complete stromal interruption was 50%. Among women with a spared cervical stroma of more than 3 mm at MR Imaging, only one had at pathologic examination microscopic metastasis in a parametrial lymphnode, accounting for a negative predictivity value of 87%. The results of our study points out a high concordance between MR Imaging findings and pathologic results, and indicate that the detection of a intact cervical stromal ring exceeding 3 mm at MR imaging is related to a very low risk of extracervical seeding of tumor. On the other side, the detection of a thinned stromal ring is related to a high incidence of parametrial invasion. The information obtained by this imaging technique may therefore be extremely useful in the accurate tailoring of treatment for these patients. PMID- 8720973 TI - [beta-Interferon in the therapy of cervico-vaginal papilloma virus (HPV) infection associated with cervical intraepithelial neoplasia (CIN)]. AB - 58 patients suffering from HPV infection associated with CIN were treated using human fibroblast interferon, via an intramuscolar route, with positive results (87.9% of cases). PMID- 8720974 TI - [Epithelial changes associated with invasive epidermoid carcinoma of the vulva]. AB - The frequency and meaning of epithelial alterations adjacent to a vulvar invasive carcinoma are examined. In the period from 1986 to 1989, 37 cases of vulvar invasive carcinoma were observed in the Bergamo Hospital (Anatomopathology Institute of Bergamo) in women aged between 54-95 years operated for vulvectomy. The vulvar epithelium adjacent to the invasive carcinoma was studied in order to determine the frequency and the type of associated lesions. Two lesions seem to be meaningfully correlated to the vulvar carcinoma: the lichen sclerosus in 59% of cases and the VIN3 of differentiated type in 35%. The identification of these epithelial lesions can serve to select the cases with increased risk to develop invasive carcinoma. PMID- 8720975 TI - [Variations in fibronectin plasma levels after surgical stress]. AB - Depletion of reticuloendothelial system following major operations or trauma closely correlates with a decrease of plasmatic levels of fibronectin. Aim of the study is to evaluate the changes in plasma fibronectin levels after a surgical stress. Twenty patients who underwent elective gynecological surgery for a variety of pathological condition were the subject of our study. Circulating levels of fibronectin showed a significative decrease, as compared to preoperative values, and returned to normal during the second postoperative day. The recovery to normal values of this plasma glycoprotein was associated, in our patients, with a normal postoperative course. PMID- 8720976 TI - [Is it possible to screen for endometrial carcinoma? Incidence and risk factors]. AB - Endometrial cancer affects 2-3% of American women. It is more common in obese, postmenopausal women with diabetes, hypertension, low parity, and late menopause. The prevalence of endometrial cancer among women who present with postmenopausal bleeding varies from 1.50-90%, most textbooks report a rate of 20%. Endometrial cancer is common, and although mortality rates relatively low, there are theoretical reasons suggesting that it could become a more serious problem in the future. The decreasing use of hysterectomy may leave more women at risk for endometrial cancer in future years. In contrast to cervical cancer, no routine mass screening programs for the early detection of endometrial cancer have been organized. It is of primary importance to define risk groups when designing mass screening programs. All of the methods used by cytologists for studying endometrial changes have limitations. Cervicovaginal smears allows diagnosis of, at best, 60% to 70% of carcinomas of the endometrium. Intrauterine sampling is another possible method. It consists of direct aspiration of the uterine cavity, scraping or brushing of the mucosa or uterine washing. Any method of endometrial sampling is imperfect, even dilatation and curettage has been shown to miss endometrial carcinoma. The quality of evidence supporting screening for endometrial cancer is of a low level. The lack of an effective, inexpensive and easy-to-use sampling method is the reason for the non-existent decline in the incidence of invasive endometrial carcinoma. PMID- 8720977 TI - [pS2 protein and breast cancer]. AB - pS2 is a small protein (7000 daltons) that is prevalently expressed in human breast cancer positive for estrogen receptors: protein is, in fact, induced by estrogen stimulation. Studies indicate that pS2 protein is a marker for hormone dependent breast tumors and that its expression is helpful in the prognosis. The presence of pS2, in fact, besides being associated in 96% of the cases to the positivities for estrogen receptors, is associated with longer overall, and disease free, survival. The pS2 protein is also expressed in normal stomach mucosa; its physiological function is unknown. PMID- 8720979 TI - Chronic recurrent pancreatitis in pregnancy. AB - This report describes a case of chronic recurrent pancreatitis due to gallstones arising in the first trimester of pregnancy. Total parental nutrition produced a normalization of pancreatic enzymes and a rapid regression of symptoms. Following another relapse of acute pancreatitis, a laparoscopic cholecystectomy was performed. The pregnancy continued normally and the patient had a spontaneous delivery at the 37th week. PMID- 8720978 TI - [Genital infections as a cause of abortion in the first trimester of pregnancy. Review of the literature]. AB - The rate of spontaneous abortion from embryo-fetal infection is believed to range from 10 to 15%. This figure, however, is likely to be underestimated because of subclinical abortions occurring at the earliest stages of gestation that go undiagnosed. The actual frequency of infections resulting in 1st trimester spontaneous abortion is therefore unknown, whereas the impact of infectious agents in the TORCH group on the pathogenesis of such event is well known, although the pathogenetic mechanisms are not well understood. A wide range of microorganisms have been isolated by several investigators from the lower genital tract of women with 1st trimester spontaneous abortion. No causative relationship of HIV-1 infection to spontaneous abortion has been established yet. The diagnosis of abortion from infection can only be made retrospectively based on histologic examination of fetal and placental tissue and isolation on culture of the infectious agent assumed to have caused infection. PMID- 8720980 TI - Neoplasms with sweat gland differentiation express various glycoproteins of the carcinoembryonic antigen (CEA) family. AB - Carcinoembryonic antigen (CEA) is a well-established marker for sweat gland differentiation in adnexal neoplasms. In contrast to previous assumptions, CEA does not represent a single oncofetal antigen but comprises a family of homologous glycoproteins, i.e. the classical CEA-180, biliary glycoprotein (BGP), and non-specific crossreacting antigens (NCA). The aim of the study was to evaluate the distribution of the respective glycoproteins of the CEA family in sweat gland neoplasms, as compared to normal sweat glands. A panel of mono specific antibodies was applied to a total of 83 samples of hyperplastic and cystic alterations of sweat glands, sweat gland neoplasms, and cutaneous metastases of different origin. Within a single group of neoplasms the immunohistochemical profile was rather consistent. Staining for both CEA-180 and NCA-90 indicated ductal differentiation of both eccrine and apocrine glands. Co expression of CEA-180, NCA-90, and BGP was consistent with differentiation towards the secretory part of eccrine glands or the transitional portion of proximal ducts. Neoplasms with signs of apocrine secretion showed a preferential immunoreactivity for NCA-90 and BGP. In conclusion, a specification of the members of the CEA family may be of some value in the differential diagnosis of adnexal neoplasms, but not in the discrimination of sweat gland carcinoma from metastatic carcinoma. PMID- 8720982 TI - In vivo expression of the insulin-like growth factor-I (IGF-I) receptor in congenital pigmented nevi. AB - Growth of normal melanocytes, nevus cells and primary melanoma cells is enhanced by insulin/insulin-like growth factor-I (IGF-I) in vitro. It has been shown that a melanoma cell line possesses the IGF-I receptor which plays a role in activation of the chemotactic response. Little is known about the in vivo expression of the IGF-I receptor and its role in melanocytic lesions. In an immunohistochemical study, we investigated the expression of IGF-I receptor in frozen sections of congenital pigmented nevi from 10 patients (ages 8 months to 4 yrs) using the monoclonal antibody alpha IR3, which specifically recognizes the extracellular alpha subunit of the IGF-I receptor. The proliferative activity of the nevus cells was examined by staining with Ki67 monoclonal antibody (reactive with all actively cycling cells). IGF-I receptor was found to be widely expressed by the cell surface of the nevus cells. Membrane staining was occasionally stronger in the superficial portion of the congenital pigmented nevi. In contrast, Ki67-positive cells were only sparsely scattered throughout the nevi with some tendency to localization to the superficial portion. This study indicates that in vivo the IGF-I receptor is widely expressed by congenital pigmented nevus cells. As opposed to keratinocytes, in which IGF-I receptor expression defines the proliferation pool of the normal and disordered epidermis, the IGF-I receptor is expressed by all nevus cells, irrespective of their proliferative status. Further studies are needed to assess whether the IGF-I receptor expression can serve as a marker for increased risk for development of malignancy in various types of benign melanocytic lesions. PMID- 8720981 TI - IgA deposition in the skin of patients with alcoholic liver disease. AB - In patients with alcoholic liver disease, IgA deposits are often found in the liver, kidneys and skin. The present study was undertaken to determine the specificity, sensitivity and characteristics of IgA deposition in the skin of a group of alcoholic patients with or without overt liver disease, and compare these with findings in non-alcoholic cirrhotics. Twenty-six out of 28 patients with alcoholic cirrhosis had IgA deposition in the skin. In contrast, only 6 out of 11 patients who were alcoholic without any clinical liver disease and 3 out of 13 patients with non-alcoholic cirrhosis of the liver had IgA deposition. In the control group, only 1 out of 52 patients with various dermatoses (excluding IgA dermatoses) had this IgA deposition. Unlike other IgA dermatoses, such as Henoch Schoenlein purpura or dermatitis herpetiformis, IgA deposition in alcoholic liver disease is characterized by its presence in the basement membrane of the eccrine secretory coils. This particular pattern of IgA deposition can be helpful in the diagnosis of alcoholic liver disease or alcoholism since the specificity is 100% with a sensitivity of 75%. PMID- 8720983 TI - Mdm-2 is not induced by p53 in human keratinocytes in vivo. AB - Normal p53 protein protects the genome after DNA damage by delaying replication and allowing DNA repair. mdm-2 is a recently discovered protein that controls p53 activity by binding to and inactivating p53 protein. A negative feedback loop has been suggested in which p53 induces mdm-2 expression. We have shown that doses of ultraviolet B radiation that cause mild sunburn clinically, produce thymine dimers in keratinocytes detectable by immunocytochemistry. This causes an elevation of p53 protein without a concomitant p53-mediated induction of mdm-2. PMID- 8720984 TI - Myoid differentiation in dermatofibrosarcoma protuberans and its fibrosarcomatous variant: clinicopathologic analysis of 5 cases. AB - We report a series of five cases of dermatofibrosarcoma protuberans, four of which were fibrosarcomatous and all of which showed histologic and immunohistochemical evidence of focal myoid/myofibroblastic differentiation (accounting for up to 5% of each tumor). These lesions were identified amongst 208 cases of dermatofibrosarcoma protuberans including 24 examples of the fibrosarcomatous variant. Three of the five patients were male and two were females; all were adults (37-72 years). One case arose on the scalp and two cases each on the abdominal wall and upper trunk. All tumors were less than 5cm in diameter and preoperative duration ranged from 2 months to 10 years. In three cases with follow-up there was no recurrence. Histologically, all tumors were typical fibrosarcomatous or ordinary dermatofibrosarcoma protuberans but for the presence of scattered to confluent nodules and bundles of eosinophilic spindle cells associated with well-defined cytoplasmic margins and vesicular nuclei associated with focal stromal hyalinization. While the typical dermatofibrosarcoma protuberans areas were CD34 positive, the myoid areas were negative for this antibody and positive for smooth muscle actin and pan-muscle actin. All tumors were desmin negative. Recognition of myofibroblastic differentiation in fibrosarcomatous dermatofibrosarcoma protuberans is important not only because it gives support to the theory of a fibroblastic/myofibroblastic line of differentiation for this type of tumor, but also because it might be a source of confusion with other myofibroblastic lesions (e.g. myofibromatosis, adult myofibroma), especially when small biopsies are evaluated. PMID- 8720985 TI - Elastic fibers in scar tissue. AB - The capacity of the skin to be stretched and to return to its resting position is correlated to the quantity and to the quality of the elastic fiber network. Although elastic fibers have been demonstrated in scars, the time course of their appearance in scars and their role in scar elasticity has not been elucidated. A study was therefore undertaken to evaluate the elastic fiber network in scars. The scars studied were from re-excision specimens following a biopsy performed for a benign or a malignant process. A total of 182 scars were evaluated in patients of different age groups. Miller's elastic tissue stain, considered to be superior to Verhoeff's van Giesen stain, was used. No elastic fibers were detected in any of 116 scars which were of less than 3 months' duration. In 66 scars present for over 3 months, a progressive increase in elastic fibers was present, first as focal and thin fibers, then as diffuse and thicker fibers. For scars of the same duration, a regional difference was noted in that scars from the back contained more and thicker elastic fibers than those from the cheek. When patients were stratified according to age, no appreciable difference was noted in the density of elastic fibers in both new and old scars between the different age groups. These results show that the synthesis of elastic tissue fibers in scars is a function of duration and site of the scar. PMID- 8720986 TI - [The morphological and morphometric study of the segment of the labyrinth of the fallopian canal]. AB - The study of the Fallopian canal with bone specimens can offer an exact and valuable information on the morphology of this duct and its close relations with neighbouring structures. The aim of this paper is to give an actualized compilation of the updated know findings, regarding the morphology and the morphometry of the first segment of the Fallopian canal. The study is based on 92 human temporal bones coming from skulls of adult subjects of both sexes. After a careful drilling work the whole length of the canal, that of the I.E.C. and the corresponding to the labyrinthine segment as well, were exposed and measured. The results allowed the AA. the macroscopical checking of the inversed ratio between the length and the calibre of the investigated canals. The labyrinthine portion of the facial canal is more narrow at its beginning (at the bottom of the I.E.C.) becoming broader as the duct approaches the first knee (geniculated ganglion) of the nerve. PMID- 8720987 TI - [Anomalous visual suppression of caloric nystagmus]. AB - In patients with central vestibular disorders it is often seen an anomaly of the nystagmus (either spontaneous or provoked) presenting as a defectuous (or non existing at all) reduction of its intensity, when the glance is fixed in a spot. We report 6 cases with this finding. Detailed clinical records, complete vestibular tests and other ancilliary studies as well, allowing to pinpoint the diagnosis, are set forth. The faulty or anomalous visual suppression is not an uniform response, because three several forms through the electronystagmographic tracins could be checked. One in which visual fixation does not affect at any the course of the nystagmic response, a second in which there is a significative reduction of the studied parameter but clearly pathologic, and the third one in which only with visual fixation nystagmus was seen. We consider that this finding indicates the existence of a central disorder, once sedative drugs are ruled out and the practice of the ENG was realized in good conditions of alertness of the patient. PMID- 8720988 TI - [WIll there ever exist the ideal auditive prosthesis for the reconstructive surgery of the middle ear?]. AB - The AA. of the paper review the several materials used for the reconstruction of the middle ear ossicular chain. They inform about each material, including the auto- and homologous grafts of ossicles, cortical bone and cartilage. Palavit, Polyethylene-90, Teflon, metallic implants. Porous plastic materials: Proplast, Plastipore and Polycel. Bioactive ceramic prosthesis (ceramics of aluminium oxyde): Frialit and CORP. Bioactive vitroceramic prosthesis: Macor, Bioglass and Ceravital. Also bioactive of phosphate of calcium: Hydroxyapatite and Apaceram. Prosthesic material of nacre, coal-coal and Polymetacrylate (Ossiculoplast). The most important matter is to get an ear in the best conditions in order to accept the prosthesis, that is to say, a bed completely dry with no inflammatory clues in the tympanic cleft and with a correct functionalism of the Eustachian tube. The healthy ossicular rennants of the chain must be spared. The apatites, bioactive ceramica, are the prosthesis of the future owing to its similitude to the bone. Nevertheless will be difficult to obtain an absolutely inert material inside the middle ear, of the stability of the cleft is not secured. PMID- 8720989 TI - [Evaluation of the cephalometric structure of the upper airway on the configuration of the obstructive sleep apnea syndrome]. AB - Thirty aleatory patients polysomnographically diagnosed as sleep apnea syndrome (SAOS) with an apnea-hypoapnea index (AHI) accounting for 61.3 +/- 31.1 were, bipedal and in decubitus, cephalometrically explored, aiming at establish the correlation between the cephalometric quantum of the upper airway and the anthropomorphic and polysomnographic parameters clearly establishing the syndrome. The weak correlation confirmed among the AHI and the back passage in decubitus (PASDEC), even between the nocturnal oxymetric nadir and the length of the palate, advice against considering the gravity of the OSAS as sole function of the altered cephalometric degree checked in the test. PMID- 8720990 TI - [Tonsillar hamartoma]. AB - Reporting the case of a young woman, 25, with long lasting dysphagia. At examination a pediculated polypoid formation in the lower pole of the left amygdala is verified (3 x 2 cms). After removal, under local anesthesia, is diagnosed as an hamartomatous tonsillar polyp. Review of the literature, analysis of several etiopathogenical theories and differential diagnosis with other resembling amygdaline conditions. PMID- 8720991 TI - [The mucociliary function and the morphology of the nasal mucous membrane in laryngectomees compared with normal subjects]. AB - The laryngectomees offer a good model for investigating the effect of absence of nasal ventilation in humans. Two samples has been studied: one composed for 24 laryngectomees, the other one for 54 healthy men. The clearance mucociliar time, using the charcoal power method in both specimen, and a structural and ultrastructural morphological survey in 5 cases, before and after the laryngectomy, was carried out. Normal people have a mucociliar clearance mean time of 9.3 +/- 3.6 minutes and among laryngectomized this time is 11.1 +/- 5.5 minutes, with no statistically significative differences at all (p = 0.26). Morphological outcome of the inquiry shows by the 2nd week there is a marked change in the previously transitional epithel which becomes a good ciliated epithelial one, together with an increase in number of goblet cells, and the exhibition of great dynamism in the morphological changes happening in the nasal mucous membrane, in agreement with the stop of the nose ventilation. PMID- 8720992 TI - [Peritonsillar abscess after tonsillectomy]. AB - The AA. report one instance of a Quinsy after some years elapsed since a tonsillectomy correctly done. No proper motive for the presentation of this phlegmonous condition could be suggested. PMID- 8720993 TI - [Clinical usefulness of the new criteria based on the serum quantification of prostate-specific antigen in the early diagnosis of prostatic cancer]. PMID- 8720994 TI - [An in-vitro study of the sources of intracellular calcium in Ach-induced contractions and electrical depolarization and of the capacity of calcium antagonists to influence them in the canine detrusor]. AB - An "in vitro" experimental line was designed to study the dependence of the canine detrussor's contractile activity on extracellular calcium, or whether on the contrary there are intracellular sources of this element. If dependence from extracellular calcium could be demonstrated, the capacity of calcium channel blockers to block the input the this ion would be studied. The conclusions arrived at were: 1. Ach- and KCl-induced activity is dependent from extracellular calcium. 2. This activity is also sensitive to the action of calcium channel blockers. 3. Calcium channel blockers block Ach-activated calcium channels completely, but only partially dose activated by KCl. 4. While the activity induced by Ach and resistant to action from calcium channel blockers is probably due to calcium released from the intracellular reservoirs, the activity induced by KCl and resistant to the calcium channel blockers action can be better explained by the activation of the transmembrane channels which cannot be blocked by this agents. PMID- 8720995 TI - [Multicentricity in renal adenocarcinoma: its incidence and the therapeutic implications]. AB - Unsuspected multicentricity in single Renal Adenocarcinoma (RAC) together with the prospects of incomplete neoplasia removal are the 2 major disadvantages for the acceptance of traditional renal surgery in birenal carriers of medium size, low grade single RAC. This paper is a retrospective review of our series of RAC patients who underwent radical nephrectomy between January 1986 and October 1994 with the following purpose: 1) To evaluate our incidence of unsuspected multicentricity. 2) To evaluate the characteristics of such multicentricity. 3) To evaluate, in the assumption that traditional surgery had been indicated for patients with small size RAC, in how many patients tumoral resection would have been incomplete due to existence of satellite tumoral nodes in the preserved renal parenchyma. Of 110 patients undergoing radical nephrectomy, 11.8% (13/110) had unsuspected multicentricity. In 10 patients there was multiple satellite nodes with size ranging from 0.3 to 3 cm. We conclude that there is not relationship between multicentricity and size or stage of primary tumour and that, by applying strict criteria (size, location, well defined tumoral wall and form of presentation) to select the patients who can be candidates to traditional renal surgery, the probability to perform it in patients with multicentric RAC would be considerably reduced. PMID- 8720996 TI - ["Ex-situ" reconstructive surgery of the renal arteries]. AB - The paper sums up our experience with kidneys which have undergone "bench" extracorporeal arterial surgery. The reasons making this procedure necessary were: renal graft from corpse donor presenting multiple arteries or arterial lesions (12 cases); orthoptic kidneys with stenotic lesions or arterial aneurism (5 cases); transplanted grafts which developed renal artery stenosis (2 cases). Incidence of arterial thrombosis and ureteral fistula was significantly higher in the first group than in grafts where no "bench" surgery had been performed. Medium- and long-term actuarial survival, however, showed no difference. In the second and third groups, extracorporeal arterial reconstructive surgery is a technique with a high risk for the loss of the involved renal unit. PMID- 8720998 TI - [Pheochromocytoma: a review of our own cases]. AB - Presentation of 6 cases of pheochromocytoma, diagnosed and treated in our Unit over the last 5 years. Five were adrenal pheochromocytomas and 1 an abdominal paraganglioma in a 42 year-old woman. Distribution by gender was 4 male and 2 female, and mean age at presentation was 45.2 years ranging from 35 to 55 years. Clinically, all patients were hypertensive. Two of the 5 cases with adrenal location presented with catecholaminic crisis with BP > 240/140 mmHg. The paraganglioma was diagnosed while studying a case of sustained HBP in a 42 year old female referred from another unit. With regard to diagnosis, the sensitivity of urinary tests was 100%, and gammagraphy with meta-iodine-benzyl-guanidine (MIBG) was particularly useful in the extra-adrenal location case. In all our patients, computerized tomography (CT) was the choice procedure to locate the tumour. Treatment was surgical in all cases, access being transperitoneal in 3 cases, thoracoabdominal in 2 and classic lumbotomy in 1. All our patients received prior treatment with alpha-blocking agents, and intraoperative complications were 1 arrythmic crisis, 1 hypotensive picture and 1 hypertensive crisis, all of which resolved successfully. Currently, 5 patients remain disease free. Mild HBP controlled with low dose captopril still persists in one patient. PMID- 8720997 TI - [Atypical forms of renal carcinoma]. AB - This is a comparative study of the various subtypes of renal carcinoma (divided according to their microscopic features in classic, papillary, Bellini's, sarcomatoid, chromophobe, small cells) on the 286 renal carcinomas removed in the Valdecilla Hospital over the last 25 years. For each type of presentation, age, gender, location, intrarenal status, tumoral diameter, clinical signs and symptoms, presence of metastasis, Fuhrman's core grade, Robson stage and survival, have been analyzed. No cases of small cells or chromophobe tumours were seen. Of the total 286 renal tumours, 253 (88.64%) showed a classical pattern, 21 (7.34%) were papillary, 10 (3.49%) sarcomatoid, and 2 (0.69%) Bellini's carcinomas. In the analysis by mean age at presentation, distribution by sex, involvement site and intrarenal location, no statistically significant differences were found between the different tumoral subtypes. In our series, the sarcomatoid type presents a larger average tumoral diameter than both the papillary and the classical ones (p < 0.01, p < 0.001, respectively). The most frequent clinical manifestation in all tumoral types was haematuria. The sarcomatoid tumours showed a statistically significant trend to present with higher core grade (G4) and higher tumoral stage (E IV) than papillary and classical tumours. Overall survival at 3 years was 72% for the papillary type, 65% for the classical and 22% for the sarcomatoid subtype, the latter being significantly worse than the first two (a < 0.001). PMID- 8720999 TI - [Ultrasound in testicular torsion]. AB - Thirty-two patients were evaluated by scrotal ultrasound prior to examination due to testicular torsion. Typical ultrasonographic patterns of testicular torsion were obtained in 100% within the first 6 hours, 25% between 6-12 hours, and 91.6% after 12 hours; the overall percentage was 84.4% for all cases. PMID- 8721000 TI - [Lumbago-sciatica syndrome as the first manifestation of metastatic renal adenocarcinoma]. AB - Presentation of one case of metastatic renal adenocarcinoma with iliac bone and soft parts involvement which presented as a lumbosciatic picture. A comment is made on the different diseases which can result in painful lumbar syndrome, reaching an agreement on the need to conduct more extensive studies given the lack of response to standard medical treatment. PMID- 8721001 TI - [Unilateral spontaneous adrenal hematoma: an unusual cause of retroperitoneal hemorrhage]. AB - Presentation of one case of spontaneous retroperitoneal haemorrhage in a 72 year old male, his first symptom being a left retrothoracic pain of sudden onset. The supplementary studies performed (ultrasound, CAT and MNR) pointed to the adrenal gland as the origin of the haemorrhage. Faced eith the uncertainty of the etiological diagnosis, a left supra-renalectomy was conducted which confirmed the pathological anatomy of the piece and the presence of massive haematoma of the suprarenal gland with no other pathological data. The clinical, diagnostic and therapeutical aspects are discussed. PMID- 8721002 TI - [Transitional-cell sarcomatoid carcinoma of the renal pelvis with osteoclast-type giant cells. Review of the literature]. AB - One case of transitional cells carcinoma with sarcomatoid pattern, located in the renal pelvis in a 77 year-old male with haematuria. The tumour also expressed a non-neoplastic component of osteoclast-like giant cells which probably represent a stromal reaction of the host to the neoplasia. The tumoral cells co-express keratin and vimentin, two strands of the cytoskeleton specific for epithelial and mesenchymal cells, respectively. We speculate with the possibility of the tumoral cells fusocellular morphology being related to the vimentin expression. A review of the literature is also included. PMID- 8721003 TI - [Renal hypoplasia and familial vesicoureteral reflux. A rare association]. AB - Presentation of two cases of unilateral congenital renal hypoplasia with vesicourethral reflux in two male siblings, no twins, both diagnosed at the age of 24. With no urological history, they present with a dysuria-pollakiuria syndrome which does not even allow to have a clinical suspicion of the final diagnosis. Clinical features of both cases are discussed, and the hyperplasia reflux association, as well as the possibility of the condition having a hereditary and familial nature, are confirmed by clinical observation as well as radiological and histological data. A literature review is undertaken but published references are very scarce. PMID- 8721004 TI - [Urocolpos: a cause of pseudo-incontinence in aged women]. AB - Presentation of our experience in three cases of urocolpos (acquired pudendal lip fusion), a series numerically equivalent to the total number of cases published until now. All patients in our series were older women and presented complete fusion of the small pudendal lips, with only a small pointed puncture in the lower part of the vulva. The main clinical signs and symptoms in our series were: urinary infection in 100%, false incontinence in 66% due to output of urine retained in the vagina, a symptom that has not been described earlier, and acute urine retention in 33%. All patients were successfully treated by means of surgical loosening of the fusioned lips and application of topical estrogens. An analysis is made of clinical and pathoanatomical features which differentiate this entity from the sclerotic and atrophic lichen. Finally, an etiopathogenic hypothesis is raised to explain the fusion acquired by the small pudendal lips in the urocolpos. PMID- 8721005 TI - [Adipose retroperitoneal tumors. Apropos a giant myxoid liposarcoma]. AB - Liposarcomas are the most frequent retroperitoneal tumours, second only to lymphomas. We present a new case of giant retroperitoneal liposarcoma treated in our hospital. A discussion is presented on the most peculiar aspects of this tumour, such us: multicentricity, tendency to local relapse, and need to post operative (adjuvant) radiotherapy. A review of all nationwide cases over the last few years is included. PMID- 8721006 TI - [Actas Urologicas Espanolas, history and spelling]. PMID- 8721007 TI - [Transcriptional regulation by cAMP and calcium]. AB - Transcription of a number of eukaryotic genes is activated in response to an increase in the intracellular cAMP concentration. These genes stimulated by cAMP have a common promoter element, cAMP response element (CRE). The CRE is recognized by a CRE binding protein, CREB. The binding of CREB to CRE does not induce transcription. Activation of transcription requires the phosphorylation or CREB at Ser-133. In the case of the cAMP pathway, the activated catalytic subunit of cAMP-dependent protein kinase (PKA) translocates to the nucleus and phosphorylates Ser-133 of CREB. In the nervous system, signals transmitted across synapses are known to regulate gene expression in the post-synaptic cell. This process often involves membrane depolarization and subsequent amplification of intracellular Ca2+. The transcriptional activation induced by membrane depolarization and Ca2+ influx is mediated by a promoter element, called the Ca(2+)-responsive element (CaRE). Recent studies of c-fos and proenkephalin gene expression have shown that the CaRE is indistinguishable from a CRE. In this paper, we focus on the possible interactions between Ca2+ and the cAMP signaling pathways into the nucleus. PMID- 8721008 TI - [Nephrotoxicity and drug interaction of vancomycin]. AB - We examined drug interactions of vancomycin hydrochloride (VCM) in the rabbit kidney. VCM has an antibacterial action against Gram positive bacteria, but composite infection patients must be jointly treated with antibiotics that are effective on Gram negative bacteria, e.g., imipenem (IPM)-cilastatin sodium (CS) compounding agent. Both VCM and IPM have the adverse reaction of nephrotoxicity, whereas CS restrains the nephrotoxicity of IPM. To clarify the interactions, we examined the nephrotoxicity and pharmacokinetics of VCM in the rabbit and compared them with those in rabbits administered VCM with CS or IPM-CS. Symptoms of nephrotoxicity such as an increase of serum creatinine concentration and BUN and a morphological change of the kidney were observed with iv. injection of VCM at 300 mg/kg. However, no abnormality of clinical data and morphological alteration were observed in the groups injected with VCM plus CS or IPM-CS. Clearance and urinary excretion of VCM obviously increased in the groups injected with VCM plus CS or IPM-CS. In addition, it was estimated that VCM was actively transported by observation of the uptake in rabbit renal slices. Furthermore, the uptake rate of VCM in the renal cortex was significantly decreased by CS. Together with the above findings, it is suggested that the restraint effect of VCM uptake into nephrocytes by CS is one of the decreasing mechanisms of the nephrotoxic effect of VCM. PMID- 8721009 TI - [Influence of KU-1257 on the recurrence and relapse of acetic acid-induced chronic gastric ulcers in rats]. AB - The influence of KU-1257 on the recurrence and relapse of acetic acid ulcers in rats was investigated grossly and histologically in comparison with that of cimetidine. The ulcer was induced by topical application of glacial acetic acid at the junction of the corpus and antrum on the anterior wall of the stomach. The drug was administered from the 5th to the 153rd day after the ulcer induction and then discontinued to the 238th day. The healing rates of the control groups (control) rose until the 119th day after the ulcer induction, followed by ups and downs. The quality of healing in the regenerated mucosa and the granulation tissue of the healed ulcer was poor, resulting in the recurrence and relapse of ulcers. The recurrence and relapse of ulcers also occurred in the cimetidine groups (CIM). On the other hand, the KU-1257 groups (KU-1257) showed much lower recurrence and relapse rates of ulcers than the control and CIM groups. Moreover, KU-1257, unlike CIM, improved the quality of ulcer healing throughout the period of its administration and even after it was discontinued. These results suggest that KU-1257 improves the quality of ulcer healing, and this may contribute to the low recurrence and relapse rates of ulcers. PMID- 8721011 TI - Iniquitous taxation of the sick. PMID- 8721010 TI - [Effect of cilnidipine on renal function in SHR and endothelin-induced renal dysfunction]. AB - Effect of cilnidipine (CIL) on renal function in SHR were evaluated in comparison with that of nifedipine (NIF) and nicardipine (NIC). In conscious SHR, CIL (3 and 10 mg/kg, p.o.) increased the urine volume, urinary Na+ excretion and urinary Na+/K+ ratio. NIF (3 and 10 mg/kg, p.o.) and NIC (10 mg/kg, p.o.) also increased the urine volume and urinary Na+ excretion, but not the urinary Na+/K+ ratio. In anesthetized SHR, CIL (3 and 10 micrograms/kg, i.v.) and NIF (10 micrograms/kg, i.v.) elevated the renal blood flow (RBF), but NIC did not. CIL (10 micrograms/kg, i.v.) also increased the glomerular filtration rate (GFR), whereas NIF did not. Furthermore, we investigated the effect of these three drugs on endothelin (ET)-induced renal dysfunction in anesthetized SHR. ET (2 micrograms/kg, i.v. +30 ng/kg/min) prolongly reduced RBF, GFR and urine volume by 47, 60 and 48%, respectively. CIL (1-10 micrograms/kg, i.v.) improved the decrease in RBF and urine volume induced by ET as well as NIF and NIC. When blood pressure was lowered to the similar extent among the three drugs in ET-treated SHR, CIL increased the RBF and urine volume compared with the others. NIF and NIC did not affect the reduction in GFR by ET, but CIL (0.3-3 micrograms/kg, i.v.) significantly increased GFR. These results suggest that CIL, NIF and NIC all have natriuretic action and no suppressive action on renal function in SHR. In addition, CIL has an ameliorative effect on renal dysfunction in ET-treated SHR, suggesting that CIL might improve renal dysfunction not only in hypertensive patients but also in those with acute renal failure. PMID- 8721012 TI - Blue ear drum. PMID- 8721013 TI - Vocal fold granuloma. PMID- 8721015 TI - Changing the nasal tip: Part IV (of VI parts). PMID- 8721014 TI - Endoscopic view of cocaine rhinitis. PMID- 8721016 TI - Immunology of skin tests. PMID- 8721017 TI - Asthma: an important disease to otolaryngologists--Part II: Asthma management strategy. AB - Asthma is a chronic inflammatory disease of the lower respiratory tract which is triggered by exposure to allergens or other airway irritants, and is commonly encountered in otolaryngologic practice. This three-part review is designed to assist otolaryngologists in effectively managing their asthmatic patients. In Part I, current information on the pathophysiology and increasing prevalence of asthma, its clinical variability, the assessment of asthma severity and methods for diagnosis of asthma were summarized. Part II discusses a tripartite strategy for asthma management, based on environmental controls, the use of anti inflammatory therapies and patient education. In addition, pharmacologic treatments which are not primarily anti-inflammatory are reviewed in detail. The uses, effectiveness, side effects and suitability for drug combination therapy for mucolytic, anticholinergic, antihistamine, theophylline and beta agonist drugs are compared, and the disadvantages of beta antagonists are summarized. Part III concludes with an overview of anti-inflammatory therapies for asthma control. PMID- 8721018 TI - Labyrinthine fistula from cholesteatoma: surgical management. AB - Labyrinthine fistula is a complication of chronic otitis media with cholesteatoma which can result in progressive sensory hearing loss or permanent loss of inner ear function. Total surgical removal of cholesteatoma and use of the intact canal wall tympanoplasty and mastoidectomy can result in preservation or improvement of hearing and elimination of vertigo. Long-term results in a series of 63 cases are reported. The important features of surgical technique involve use of high magnification with the operating microscope, a high-speed drill, suction irrigation and thorough knowledge of temporal bone anatomy. In addition, the surgeon must avoid tearing or perforation of the squamous epithelium basement membrane during dissection. In some cases, sensory hearing returns to normal following surgery, making a successful second-stage reconstruction for hearing possible. PMID- 8721019 TI - Tympanoplasty in adults: a five-year survey. AB - In order to identify factors that could influence the success of tympanoplasties in adults, a retrospective study of 291 tympanoplasties performed on patients aged 15 years and older during the years 1984-1989 was carried out. Surgical success was defined as an intact membrane after two years follow-up and hearing improvement of at least 15 decibels. The success rate of the tympanoplasties was 90%. A functioning eustachian tube was found to be important for graft take. No difference was found in the rate of graft take regardless of whether fascia of the temporal muscle or tragal perichondrium was used or if fibrin glue was used. An audiological improvement was seen in 77% of the patients, and no significant difference regarding hearing gain was found between the different prostheses. It is concluded that tympanoplasty in adults is a simple operation with a high probability of success which can improve the quality of life of those operated upon. PMID- 8721020 TI - Facial nerve palsy as an early manifestation of relapse in T-cell acute lymphoblastic leukemia. AB - We describe a 14-year-old girl with T-cell acute lymphoblastic leukemia (ALL) who developed peripheral type facial palsy alternately on each side during remission. The cerebrospinal fluid examination and imaging studies of the brain including the temporal area failed to identify the etiology. Fully 10 weeks after the onset of the facial nerve palsy, she showed cytological findings of meningeal and bone marrow relapse. Review of the literature revealed that an isolated facial nerve palsy due to leukemic cell infiltration is a rare occurrence. Nonetheless one needs to keep in mind that it may represent the initial sign of a relapse in a patient with leukemia, particularly with T-cell ALL. PMID- 8721021 TI - Neurogenic stuttering: further corroboration of site of lesion. AB - Cases of neurogenic stuttering have been reported in the literature throughout the past century. Site(s) of lesion(s) have been documented usually by association of symptoms, EEG studies and occasionally by computed tomography (CT). The authors present three cases in which the site(s) of lesion(s) are documented by CT, magnetic resonance imaging (MRI) and SPECT. This study supports previous findings of neurogenic stuttering following either bilateral diffuse lesions or a unilateral lesion. In at least one case, the actual site of the lesion would have been missed without the use of SPECT testing. EEG studies were not helpful in identifying the site of the lesion. PMID- 8721022 TI - Orbital floor fracture following nose blowing. AB - A 70-year-old woman with an upper respiratory infection sustained an orbital floor fracture after vigorous nose blowing. The injury was complicated by orbital emphysema and cellulitis. CT scanning confirmed orbital floor fracture and associated orbital soft tissue herniation into the maxillary antrum. A follow-up sinus endoscopy three months later showed a healed periosteum and mucosa, but continued orbital herniation. As the patient's symptoms had largely resolved, repair was averted. Follow-up after nine months showed no recurrence of orbital swelling. The case presented illustrates a rare complication of vigorous nose blowing, coupled with thinning of the sinus walls seen in aging. PMID- 8721023 TI - Extramedullary nasal plasmacytoma--an unusual clinical entity. AB - A case of extramedullary plasmacytoma with its unusual appearance is reported. This is the second reported case in world literature affecting the paranasal sinuses with intracranial extension. The role of surgery is to obtain tissue for diagnosis and to excise residual disease. Radiotherapy is the treatment of choice and long-term follow-up is necessary for monitoring disease recurrence. The overall 10-year survival is about 50%. The case is discussed with a general review of the management of this pathology. PMID- 8721024 TI - Genetic nomenclature: ISCN and ISGN. PMID- 8721026 TI - Burkitt's lymphoma in 63 Turkish children diagnosed over a 10 year period. AB - Sixty-three Turkish children with Burkitt's lymphoma (BL) diagnosed over a 10 year period in a single institution were retrospectively analyzed. Burkitt's lymphoma included 41.7% of non-Hodgkin's lymphomas and 17.2% of all childhood malignant solid tumors diagnosed in our department in this duration. The patients studied with BL were aged between 3 and 14 years (mean 5.9 years), with a male of female ratio of 2:1. While the age distribution in our patients was similar to that in African BL (endemic), the predominance of abdominal involvement and the frequency of bone marrow infiltration and pleural effusion were reminiscent of American BL (sporadic). The incidence of jaw involvement (15.9%) in our group was higher than in American BL, however, and was not a high as in African BL. Most of the patients were of a lower socioeconomic status. Significant growth retardation was found in the children with BL compared with 40 age-matched children without malignancy, nor chronic or endocrinologic disorders, who were of a similar socioeconomic status. A serological study for Epstein-Barr virus (EBV) was performed in 18 children, and the IgG-type antibody to the viral capsid antigen of EBV was found to be positive in all of them. As a result, BL seems to include a considerable proportion of all childhood malignant solid tumors in Turkey. The epidemiological and clinical presentation and course indicate that BL appears in Turkish children in a form that is between the African and American types of the disease. Further molecular and chromosomal studies in Turkish children with BL are needed. PMID- 8721025 TI - Treatment with recombinant human erythropoietin in children with malignancies. AB - The effect of recombinant human erythropoietin (rHuEPO) on the anemia of cancer was examined in 15 children with hematologic malignancies (group I) and solid tumors (group II), whose hemoglobin (Hb) was under the third percentile for sex and age. The response to rHuEPO was defined as an increase of Hb to above the 10th percentile following 8 weeks of therapy. The rHuEPO caused an increase in the Hb and hematocrit (Hct) in 46% of children of both groups at a dose of 150 IU/L, in 28.5% of children at a dose of 250 IU/L and in 25.5% of children at a dose of 400 IU/L. Leukocyte and platelet counts were not influenced by the rHuEPO treatment. The red cell transfusion requirement decreased to 66% in both groups after rHuEPO treatment. Erythropoietin (EPO) levels were measured prior to the treatment and then every 4 weeks during rHuEPO treatment. Children who responded to EPO had an initial EPO level of < 100 IU/L, while those who did not respond had an initial EPO level of > 100 IU/L. Erythropoietin was well tolerated in all children, with no side effects. PMID- 8721027 TI - Febrile episodes in children with cancer in the United Arab Emirates. AB - The febrile episodes encountered in our pediatric oncology unit over a 2-year period were reviewed. A total of 138 febrile episodes were recorded in 59 patients (29 with leukemia and 30 with a solid tumor). There was no difference in the number of episodes between leukemia and solid tumor patients, nor between neutropenic and non-neutropenic patients. The degree of neutropenia was more severe in leukemia patients. A total of 18.8% of the episodes were accompanied by positive blood cultures. Gram-positive bacteria were more frequent than gram negative bacteria, and there were four anaerobic isolates. Seventeen episodes were accompanied by clinical signs of central venous line (CVL) infection. A total of 70.2% of the episodes resolved with a first-line antibiotic combination of flucloxacillin, piperacillin, and netilmicin, 27.5% required modification of the antibiotic combination, and three patients (5%) died due to gram-negative septicemia. These findings indicate that the pattern of infectious complications in the United Arab Emirates is now similar to that observed in Europe and the United States. PMID- 8721028 TI - Infection-associated hemophagocytic syndrome complicated by infectious lymphoproliferation: a case report. AB - The case of a 7-year-old boy with virus-associated hemophagocytic syndrome (VAHS) and serologically proven parvovirus B-19 infection is described. The patient with VAHS presented with fever, hepatosplenomegaly, pancytopenia, and hyperlipidemia type IV. After induction therapy with VP-16 and prednisone, partial remission was achieved. Despite maintenance therapy, reinductions, and the addition of cyclosporine A for 3 months, several relapses occurred. The therapy was stopped because of life-threatening complications (Klebsiella sepsis, neutropenic enterocolitis, and stercoral peritonitis). The complications were treated successfully. The patient status was stabilized after splenectomy. However, hepatomegaly progressed slowly and the hyperlipidemia endured. Ten months after the diagnosis leukocytosis with absolute T lymphocytosis appeared. Reactivation of VAHS was suspected and intravenous immunoglobin and then antilymphocyte immunoglobulin ALG therapy were started. The resultant decrease in leukocytosis was prompt, but lymphopenia did not occur. Virostatic treatment with foscarnet was introduced based on human herpesvirus-6 seroconversion. Twenty-six months after the diagnosis, the patient is well, without any sign of VAHS or lymphoproliferation. PMID- 8721029 TI - Transient acute monoblastic leukemia with reciprocal (8;16)(p11;p13) translocation. AB - The translocation of t(8;16)(p11;p13) has been demonstrated in the blasts of a phenotypically normal newborn baby with acute monoblastic leukemia. No antileukemic therapy was administered and spontaneous, complete remission was observed at 2 months of age. The patient remains well 18 months after the diagnosis and continues to have a normal hemogram. PMID- 8721030 TI - Etoposide in Langerhans cell histiocytosis in children: a preliminary experience. AB - Treatment of Langerhans cell histiocytosis (LCH) is yet to be established. We treated seven patients with etoposide alone at a dose of 100 mg/m2/day for 3 days given every 3 to 4 weeks for six cycles. Three patients had received prior chemotherapy, two patients were less than 2 years of age, and two had liver dysfunction. A positive response to therapy was seen in five patients. There was no major toxicity. Etoposide therapy is safe and effective in the treatment of LCH. PMID- 8721031 TI - Prenatal diagnosis of hemoglobinopathies in Turkey: Hacettepe experience. AB - Prenatal diagnosis of hemoglobinopathies was performed in 250 fetuses at risk for hemoglobinopathies. The main diagnostic procedures were in vitro hemoglobin synthesis analysis in fetal blood and analysis of DNA obtained from chorionic villus samples. Sixty-six percent of the fetuses were at risk for beta thalassemia major and 28% for sickle cell anemia. Beta thalassemia mutations were heterogenous, and 51 fetuses examined by the DNA technique were found to be at risk for at least 20 different combinations. PMID- 8721032 TI - An adolescent case of retroperitoneal pure choriocarcinoma: successful treatment with MCNU-containing high-dose chemotherapy followed by autologous bone marrow transplantation after multiple brain metastases. AB - Choriocarcinoma is a rare disease in pediatric neoplasms. The prognosis of the disease is extremely poor once when patients relapse or become refractory to cisplatin (CDDP). A 17-year-old male who had retroperitoneal pure choriocarcinoma of advanced stage was treated with CDDP-based intensive chemotherapy. In spite of the initial good response to CDDP-based intensive chemotherapy, the tumor metastasized to multiple areas of the brain during chemotherapy. Since the brain in this case was thought to be a sanctuary, after radiotherapy to the whole cranium, the patient was treated with high-dose chemotherapy consisting of etoposide, carboplatin, and ranimustine (MCNU), which can penetrate the blood brain barrier, followed by autologous bone marrow transplantation (ABMT). Twenty four months after ABMT, the patient had no sign of disease recurrence. MCNU containing high-dose chemotherapy with ABMT appears to be quite effective in cases that present with relapsing multiple brain metastases during CDDP-based chemotherapy. PMID- 8721033 TI - Transient aplastic crisis in a leukemic child caused by parvovirus B19 infection. AB - Acute parvovirus B19 infection was confirmed in an 8-year-old girl with acute lymphoblastic leukemia on maintenance therapy by electron microscopy, DNA, and serological techniques. Clinical manifestations were typical of severe anemia, fever, leukopenia, and thrombocytopenia. Increased levels of serum erythropoietin were detected in the course of the infection. The patient's recovery was supported by corticosteroids, red blood cell transfusions, G-CSF, and commercial immunoglobulin preparations. PMID- 8721034 TI - Suspicion of child abuse complicating the diagnosis of bleeding disorders. PMID- 8721035 TI - A case of severe thrombocytopenia due to parvovirus B19 virus. AB - A 9-year-old girl admitted with generalized skin, mucosa, and genitourinary system bleeding had anemia and thrombocytopenia. Her bone marrow was normocellular. Parvovirus B19 IgM and IgG were positive. An absence of reticulocytopenia suggested that the virus affected only the megakaryocytic cell line and the anemia was due to generalized bleeding resulting from thrombocytopenia. Intravenous immunoglobulin treatment was instituted. On the tenth day of hospitalization the patient recovered from anemia and thrombocytopenia. IgM antibodies disappeared. PMID- 8721036 TI - Dramatic effect of high-dose methylprednisolone on orbital granulocytic sarcoma. PMID- 8721037 TI - Successful multidrug chemotherapy of olfactory neuroblastoma in an 8-year-old child. PMID- 8721038 TI - Rothmund-Thomson syndrome, malignant diseases, and treatment opportunities. PMID- 8721039 TI - Myelodysplastic syndromes and RTS. PMID- 8721041 TI - A rapid approach for the detection of dipicolinic acid in bacterial spores using pyrolysis/mass spectrometry. AB - Curie-point pyrolysis/triple quadrupole mass spectrometry and micro-tube furnace pyrolysis/quadrupole ion trap mass spectrometry have been used to detect dipicolinic acid (DPA) in sporulated whole bacteria. DPA in whole cells of sporulated Bacillus anthracis reacted in situ during pyrolysis with tetramethylammonium hydroxide to form the dimethyl ester derivative of DPA, dimethyl-2,6-dipicolinate (mDPA). The mDPA was identified by its positive-ion electron ionization fragmentation pattern and confirmed with tandem mass spectrometry. In an oxidative pyrolysis/quadrupole ion trap instrument, the mDPA mass spectrum showed characteristic positive-ion electron ionization fragmentation along with a significant [M+1]+ ion due to self-chemical ionization. The characteristic collision-induced dissociation fragments of mDPA were used to establish the presence of sporulation in B. anthracis whole cells at a concentration of 2.2 x 10(7) CFU (colony-forming units)/mL using the triple quadrupole instrument. The total time for analysis, including sample preparation, was less than 10 minutes for both instruments. PMID- 8721040 TI - A randomized study of ondansetron syrup in children: evaluation of taste acceptability and tolerance. AB - Ondansetron, a highly selective 5-HT3 receptor antagonist, is available in an intravenous (IV) formulation and tables, but syrup would be particularly useful in children. As chemotherapy can affect taste perceptions, this study was undertaken to determine the preference between two flavors of ondansetron syrup in children undergoing chemotherapy. Fifty-nine children, randomized into a multicenter, double-blind, crossover study, each received 5 mg/m2 of IV ondansetron daily before chemotherapy. The syrup was then randomly given in two doses, one of each flavor, strawberry and grape, 30 minutes apart. The preference was assessed 30 minutes after the second dose of syrup had been administered. Taste was assessed by the child against a panel of five faces. Of those children expressing any preference, 70% preferred the strawberry flavor. Overall, 59% of children preferred the strawberry flavor, whilst 25% preferred grape (P = 0.005) and 15% expressed no preference. The only adverse event assessed as drug related by the investigator was constipation, which occurred in one patient. In conclusion, a strong preference was found for the strawberry formulation. The ondansetron syrup was safe and well tolerated. PMID- 8721042 TI - Comparing mass spectrometric characteristics of peptides and peptoids. AB - The collision-induced dissociation (CID) spectra of the [M+H]+ ions of a pentapeptide and the corresponding peptoid and retropeptoid have been compared. The spectra of the peptide and peptoid both exhibit B- and Y"-type sequence ions at identical m/z values. In contrast to the peptide, the [M+H]+ ion of the peptoid and all sequence ions containing an N-substituted glycine derivative corresponding to a tyrosine amino acid residue can easily lose a C7H6O molecule in a charge-remote fragmentation process. The presence of N-substituted glycine residues in a peptoid is further apparent from the presence of N-substituted immonium ions, which differ significantly in their fragmentation behaviour from the corresponding immonium ions observed in the spectra of common oligopeptides. Loss of the CH2 = NH imine molecule is the dominant fragmentation reaction in the CID spectra of all peptoid immonium ions investigated in this study. The elimination of the CH = NH2 ylide analogue from common peptide immonium ions is energetically less favourable as shown by ab initio calculations. The relative heat of formation of the CH = NH2 ylide neutral appeared to be 168 kJ mol-1 more than that of the CH2 = NH imine molecule. PMID- 8721043 TI - Immunosuppression in tuberculosis. PMID- 8721044 TI - The role of T-cell subsets in Mycobacterium tuberculosis infection. AB - During the last 10 years, studies of the immune response to M. tuberculosis in humans and animal models have increased our understanding of the complex roles of T-cell subsets in protection against tuberculosis. Although CD4+ T cells remain the dominant and critical T-cell subset, others, such as gamma delta and CD8+ T cells, probably have important complementary roles. Since all three subsets are sources of IFN-gamma and competent cytotoxic effector cells, in vivo kinetics and differences in antigen processing/recognition likely will define how each T-cell subset functions in different phases of the immune response to M. tuberculosis. In addition, individuals may differ in terms of the dominance of CD8+ and gamma delta T cells as accessory T-cell populations. In some, gamma delta T cells; in others, CD8+ T cells; or both T-cell subsets may complement CD4+ T-cell function. Future studies in animal models (with human cells obtained from sites of infection such as lung or lymph node), characterizations of the antigen repertoire, and longitudinal immunoepidemiological studies should define more clearly how different T-cell subsets contribute to protection against M. tuberculosis. Such studies may determine how failure of T-cell subset function results in reactivation or progressive primary tuberculosis. Enhanced understanding of the function of, and antigen recognition by, T-cell subsets in M. tuberculosis infection also is necessary for the development of improved diagnostic tests and vaccines for tuberculosis. PMID- 8721045 TI - Regulation of the human immune response in tuberculosis. PMID- 8721046 TI - Cytokine circuits in tuberculosis. PMID- 8721047 TI - Pulmonary immune response to Mycobacterium tuberculosis and human immunodeficiency virus. PMID- 8721048 TI - New approaches to identification of antigens of Mycobacterium tuberculosis. AB - The antigenic repertoire presented by mycobacteria is a complex mixture of protein and nonprotein antigens. Responses to these antigens may lead to elaboration of cytokines which are important in the development of protective immunity or, conversely, may be involved in immunopathogenesis. Successful development of subunit vaccines for TB will depend on identification of specific antigens or epitopes which can selectively activate mechanisms involved in protective immunity. Development of these vaccines will require preparation of relatively large volumes of LAM-free antigens for testing in animal models of TB which most closely represent human disease. PMID- 8721049 TI - [A study on the maternal plasma C21 steroids concentration during pregnancy]. AB - During pregnancy, C21 steroids such as progesterone (P4), and cortisol (F), have been reported to be closely involved in uterine contraction. To clarify the association between changes in steroid hormones and the onset of delivery, we measured the concentrations of six C21 steroids in maternal blood by high performance liquid chromatography. Changes in each steroid hormone were evaluated by analysis of variance during pregnancy. Pregnenolone, a source of C21 steroids, gradually increased during pregnancy. P4 and 17P4, its metabolite, reached a peak 3 weeks before delivery and noticeably decreased thereafter. Accompanying a decrease in P4 and 17P4, 20P4 and F, its metabolite, were noticeably increased. The association among these steroid hormones was also evaluated. The correlation between P4 and F was reversed after 3 weeks before delivery. These results suggest that steroid hormones in the maternal blood begin to change dynamically about 3 weeks before delivery. In particular, a decrease in P4 and an increase in F seem to be closely related to the onset of delivery. PMID- 8721050 TI - [Effects of basic fibroblast growth factor on the development of mouse preimplantation embryos]. AB - Basic fibroblast growth factor (bFGF) is known to be a mesoderm inducer of the Xenopas embryo, although the role of this factor in mammalian preimplantation embryos is not known. This study was performed to examine possible roles of bFGF in murine preimplantation development. To determine the expression of bFGF and FGF receptor 1 (FGFR 1) mRNA in mouse embryos and uterine endometrial epithelial cells, a reverse transcription-polymerase chain reaction (RT-PCR) technique was used. In the mouse embryos, bFGF mRNA was not detected but FGFR 1 mRNA was expressed in the blastocyst stage. Long and short forms of FGFR1 mRNA generated by alternative splicing were expressed. Both bFGF and two forms of FGFR 1 mRNA were detected in mouse endometrial epithelial cells. Immunoblot analysis indicated that bFGF protein was present in the uterine luminal fluid during the preimplantation period, and the level of expression of the protein was relatively constant. The addition of bFGF to the culture medium had no effects on the rate of blastocyst formation of 2 cell stage embryos, but it significantly increased the protein synthesis in blastocysts. These results suggest that bFGF derived from the mouse uterine endometrium affects the development of preimplantation embryos in a paracrine fashion. PMID- 8721051 TI - [Changes in serum lipoprotein (a) levels related to hyperlipidemia during pregnancy--comparing normal pregnancy and toxemia of pregnancy]. AB - In this study, we investigated fluctuations in serum lipoprotein (a) (Lp(a)) levels in normal and toxemic pregnancy. We measured serum total cholesterol (TC), triglyceride (TG), phospholipid (PL), high-density lipoprotein (HDL), apolipoprotein and Lp(a) levels in 33 normal pregnant and 11 toxemic pregnant women at delivery and in 47 normal pregnant women throughout gestation. Lp(a) and apolipoproteins were detected by turbidimetric immunoassay. The levels of serum lipids, HDL and apolipoproteins were all increased in pregnancy. In toxemia of pregnancy, serum TC and PL levels were lower (p < 0.05) and the apolipoprotein C III level was higher (p < 0.005) than in normal pregnancy. Serum Lp(a) levels increased until the 20th week and reached a value which was 1.5 times higher than at the 10th week. Thereafter Lp(a) levels were constant until the late stage of pregnancy. In contrast, serum TC and TG levels increased steadily throughout gestation. The serum Lp(a) level was 18.1 +/- 27.5mg/dl in normal pregnancy and 17.8 +/- 17.9mg/dl in toxemia of pregnancy. These results revealed changes in serum Lp(a) levels during pregnancy. Further studies will be required to clarify the metabolic control of Lp(a) in pregnancy and the matabolic disorders of lipids and lipoproteins in toxemia of pregnancy. PMID- 8721052 TI - [Significance of middle cerebral artery doppler velocimetry based on behavioral states in intrauterine growth retarded fetuses--prediction of development of fetal distress]. AB - Middle cerebral artery resistance indices (MCARI) based on fetal behavioral states were calculated in 60 normal and 20 intrauterine growth retarded (IUGR) fetuses between 33 and 40 weeks of gestational age. Fetal behavioral states were determined by recording fetal actocardiograms. The results obtained were as follows: 1. MCARI was significantly lower in the active phase than in the resting phase in normal fetuses. 2. Of the 20 IUGR fetuses, 9 subsequently developed fetal distress. Seven of 11 fetuses who did not develop fetal distress had abnormally low MCARI (less than M-2.0 SD) in the active phase. On the other hand, seven of the fetuses that subsequently developed fetal distress had abnormal MCARI not only in the active phase but also in the resting phase. The fetuses that did not develop fetal distress had different MCARIs in the two phase, but the fetuses that developed fetal distress exhibited no such difference. 3. The sensitivity and specificity in predicting the development of fetal distress in IUGR fetuses with abnormal MCARI in the resting phase were 77.8% and 81.8%, respectively. This study showed that in normal fetuses MCARI differs according to the behavioral state, and that resting phase MCARI is useful for predicting fetal distress in IUGR fetuses. PMID- 8721053 TI - [Analysis of the mechanism of trophoblast infiltration]. AB - A cytotrophoblast (CT) infiltrates into the stroma, forming an extravillous trophoblast (EVT) in the placenta early in gestation and the phenomenon is strictly controlled, differing from the infiltration of cancer cells. The expression of matrix metalloproteinase 2 (MMP2) and matrix metalloproteinase 9 (MMP9), which deeply involve infiltrative metastasis of cancer, and the reactivity to transforming growth factor beta 1 (TGF beta 1), which controls the expression of these MMPs and inhibits the growth of epithelial cells, were investigated in CT derived from villi at normal gestational week 6 (early CT) and CT derived from villi at normal gestational week 37 (full-term CT), and also the choriocarcinoma cell line BeWo (BeWo). The ability of normal epithelial cells and BeWo cells to proliferate and infiltrate were evaluated in vitro by northern blotting, gelatin zymography, and invasion assay. It was revealed that early CT had a higher capacity for infiltration than full-term CT as well as BeWo. MMP2 and MMP9 appeared in the early CT, whereas only MMP9 was observed in the full term CT. MMP2 and MMP9 were more abundantly observed in the early CT and the full term CT rather than in BeWo. In uterine stroma-derived cells, membrane type matrix metalloproteinase (MT-MMP), which activates MMP2, was observed. These results indicated that the motility of normal villous cells was higher in the early CT than in the full-term CT. The expression of MMP2 in the early CT, which was not observed in the full-term CT, was thought to be related to this difference in motility. As for the responsiveness to TGF beta 1, which is a growth inhibiting factor for epithelial cells, the villous carcinoma cell line was insensitive to the growth inhibiting effect of TGF beta 1, but the early CT was sensitive to this effect. When TGF beta 1 was added, MMP2 and MMP9 increased in the early CT. This response was also seen in BeWo. That is, it was assured that the growth capacity was not inhibited in BeWo, but was certainly inhibited in the early CT. The overall results of these evaluations indicated that the development to EVT by infiltration of the early CT was associated with the increase in the mobility of cells caused by MMP2 and the increase in amounts of MMP2 and MMP9 caused by TGF beta 1, and the predominant inhibitory effect of TGF beta 1 on the growth of normal epithelial cells could explain why normal epithelial cells do not grow as cancer cells do. PMID- 8721054 TI - [System for registering gestational trophoblastic disease in Chiba Prefecture in the past 20 years]. AB - From 1974 to 1993, a total of 3,203 patients with hydatidiform mole (HM), were registered in Chiba Prefecture. The incidence of HM during the study period was 2.8 per 1,000 live births or one per 357 live births, but it decreased to 1.5 per 1,000 live births in these 3 years. In the past 20 years, the incidence of invasive mole (IM) and choriocarcinoma was 8.8% (282/3,203) and 0.91% (29/3,203), respectively. In addition, 1,599 patients with HM have been diagnosed and registered as complete mole (CM) and partial mole (PM) patients on the basis of macroscopic findings since 1981. Of these classified patients, the incidence of IM following CM and PM was 12.5% (141/1,130) and 1.5% (7/469), respectively. Moreover, 19 (1.7%) patients with CM and one (0.2%) patient with PM developed CC. In this study, it is clear that gestational trophoblastic disease after the evacuation of PM definitely occurred and the management of PM should be similar to that of CM so long as macroscopic criteria are employed. PMID- 8721055 TI - [Combined effects of sizofiran and rG-CSF on myelosuppression in cancer chemotherapy]. AB - A combination of the antitumor polysaccharide Sizofiran and rG-CSF was investigated for its effects on peripheral blood counts in ovarian cancer patients undergoing chemotherapy. We performed the analysis by using variations in peripheral blood counts and the duration of treatment as variables. In the analysis, effects on decreases in both neutrophils and platelets and their recoveries could be assessed. As a result, the decrease in neutrophils was inhibited and its recovery was promoted, suggesting that the combined use of the drugs may be useful for myelosuppression by chemotherapy for ovarian cancer. PMID- 8721056 TI - [Feasibility of peritoneal washing cytology through the totally implanted reservoir for early detection of peritoneal recurrence of ovarian cancer]. AB - There have been no reliable ways to follow up ovarian cancer patients receiving optimal debulking. In this study, we assessed the feasibility of cytological diagnosis on peritoneal washing cells collected through a totally implanted reservoir as a tool in following up these patients. Between 1988 and 1993, 31 patients underwent optimal debulking and were implanted with a reservoir. After the operation, the patients were followed up at 2-4 week intervals with peritoneal washing cytology. Six patients were diagnosed as having a recurrence by the peritoneal washing cytology alone. And at this point there had been no other positive findings by other conventional methods. There were no complications caused by reservoirs. Activated mesothelial cells, multinucleated histiocytes and hemosiderine-laden histiocytes were characteristic cytologic findings accompanying adenocarcinoma cells. Hemosiderin-laden histiocytes were found by peritoneal washing cytology even before the appearance of adenocarcinoma cells. The appearance of hemosiderin-laden histiocytes may therefore be a sign of intraperitoneal recurrence of ovarian cancer. CONCLUSION: 1) Peritoneal washing cytology through a reservoir is useful for early detection of intraperitoneal recurrence of optimally debulked ovarian cancer without any significant complications. 2) The appearance of intraperitoneal hemosiderin-laden histiocytes may be sign of intraperitoneal recurrence of ovarian cancer. PMID- 8721057 TI - [Utility of predeposit autologous blood donation by switch back method combined with recombinant human erythropoietin in gynecological surgery]. AB - To apply recombinant human (rh)erythropoietin (EPO) to predeposit autologous blood donation (P.A.B.D.) in cancer patients clinically, (1) we tested the effects of rh-EPO on 2 ovarian cancer cell lines in vitro at first, then, (2) studied the effect of the rh-EPO for switch back (SB) method that is a variant of P.A.B.D. clinically. Rh-EPO (0.068-68U/ml) caused no significant and reproducible stimulation of clonal growth to SHIN-3 (derived from serous cyst adenocarcinoma) and MN-1 (derived from mucinous cyst adenocarcinoma). Twenty-five cases were studied. The change in the hemoglobin concentration (delta Hb) was -0.43 +/- 1.38g/dl (mean +/- SD) and the change in the total amount of hemoglobin (total delta Hb) which is calculated on the basis of whole blood volume was 111.5 +/- 53.2g/body in 16 cases with rh-EPO. The delta Hb and total delta Hb were -3.25 +/ 0.78g/dl and 30.1 +/- 41.7g/body in 9 cases without rh-EPO. The rh-EPO combined cases were significantly increased in both delta Hb and total delta Hb (p < 0.05, unpaired student t test). We therefore conclude that it would be very beneficial to use rh-EPO combined with the SB method in P.A.B.D. for high maximum surgical blood order schedule (MSBOS) cases such as gynecological malignancies. PMID- 8721058 TI - [DNA diagnosis for short limb dwarfism, possibility of prenatal diagnosis for achondroplasia]. PMID- 8721059 TI - [A case report: preoperative diagnosis of placental site trophoblastic tumor]. PMID- 8721060 TI - [Two cases of agnathia-holoprosencephaly complex]. PMID- 8721061 TI - [Limitation of uterine weight in total vaginal hysterectomy in patients with uterine myoma and adenomyosis]. PMID- 8721062 TI - [An investigation on the therapeutic effects of tuboplastic catheter system under salpingoscope (FT catheter system) for the patients with bilateral tubal occlusion]. PMID- 8721063 TI - Videomanometric aspects of pharyngeal constrictor activity. AB - Pressure changes were registered with videomanometry (simultaneous manometry and barium swallow) in the pharynx and in the pharyngoesophageal segment (PES) during swallowing. A considerable longitudinal asymmetry was found. Peak pressure was highest in the PES, lower in the inferior constrictor area, and lowest at the level of the tongue base. The rate of pressure rise was highest at the level of the PES. The speed of propagation of the contraction wave was 13 (+/- 2) cm/sec. There was no correlation between the measured variables (i.e., peak pressure, rate of pressure rise, and speed of contracting wave). Our findings can partly be explained by different mechanical constraints at different levels of the pharynx but may also reflect the organization of neural control of swallowing in the brainstem. Knowledge of transducer position and orientation is essential for the evaluation of pharyngeal pressure during swallowing. Such knowledge is best achieved by performing manometry simultaneously with fluoroscopy, i.e., videomanometry. PMID- 8721064 TI - Prevalence of gastroesophageal reflux in patients who develop pneumonia following percutaneous endoscopic gastrostomy: a 24-hour pH monitoring study. AB - Percutaneous endoscopic placement of feeding gastrostomies (PEG) was pioneered by Gauderer et. al. in 1980 [1]. Since then, it has become the preferred method of providing enteral nutritional support in children and adults because of advantages in morbidity and cost [2,3]. Pneumonia is a known sequel of this procedure, occurring at different rates, depending on the length of follow-up. Some series have shown an incidence of 10% at 30 days and others 56% at 11 months [4,5]. It does not appear that PEG feeding offers an advantage over the more traditional naso-enteric tube feeding methods in this respect. To study the prevalence of gastroesophageal reflux (GER) in PEG-fed patients, we quantitated GER by 24-hour intraesophageal pH monitoring in a group of patients who developed post-PEG pneumonia and compared it with a control group. Our study demonstrates an increased prevalence of GER in the pneumonia group compared with the control group. However, the exact contribution of this observed increased GER to the development of pneumonia needs to be determined. PMID- 8721065 TI - Mechanisms of gastric distention in esophageal atresia with distal tracheoesophageal fistula. AB - This paper explains how the chest of the crying infant with the common form of esophageal atresia is a pump for ventilating the distal esophagus and inflating the stomach. Esophageal inflation from the trachea occurs during inspiration and esophageal deflation into the stomach during cry. PMID- 8721066 TI - A penetration-aspiration scale. AB - The development and use of an 8-point, equal-appearing interval scale to describe penetration and aspiration events are described. Scores are determined primarily by the depth to which material passes in the airway and by whether or not material entering the airway is expelled. Intra- and interjudge reliability have been established. Clinical and scientific uses of the scale are discussed. PMID- 8721067 TI - The significance of accumulated oropharyngeal secretions and swallowing frequency in predicting aspiration. AB - This study retrospectively investigated the value of both endoscopically visible oropharyngeal secretions in the hypopharynx and swallowing frequency in the prediction of aspiration of food and liquid. Fiberoptic endoscopic evaluation of swallowing (FEES) was performed on a total of 69 individuals that included hospitalized elderly, nonhospitalized elderly, and young normal subjects. A four level rating scale for determining the severity of accumulated oropharyngeal secretions was developed and employed to rate subjects prior to the presentation of food or liquid during the FEES. Spontaneous dry swallows were also counted during the observation period of the FEES. It was found that the accumulation of endoscopically visible oropharyngeal secretions located within the laryngeal vestibule was highly predictive of aspiration of food or liquid. There were significantly fewer spontaneous swallows in hospitalized subjects when compared with nonhospitalized subjects. There was also a significant decrease in the frequency of spontaneous swallows in aspirating hospitalized subjects when compared with nonaspirating hospitalized subjects. Results are discussed in terms of integrating this information with clinical bedside examination techniques. PMID- 8721068 TI - Prandial aspiration and pneumonia in an elderly population followed over 3 years. AB - The purpose of our study was to prospectively determine pneumonia frequency and correlate it with prandial liquid aspiration and feeding status in frail elderly nursing home residents. Initially, 152 patients had video swallowing examinations (81 oropharyngeal dysphagia, 19 thoracic dysphagia, 52 without dysphagia). Those diagnosed with oropharyngeal impairment were subsequently managed with swallowing therapy or artificial feeding modalities. Patients were followed for 3 years (unless they expired earlier) and clinical courses were categorized according to the degree of prandial aspiration and feeding (PAF) status. Subjects with new lung infiltrates persisting for at least 5 days with appropriate clinical findings were diagnosed as having pneumonia and were classified according to the PAF status months in which these findings occurred. Fifty-six pneumonias were diagnosed during 4,280 months with the following frequencies: no aspiration months 0.6%; minor aspiration months 0.9%; major aspiration/oral feeding months 1.3%; major aspiration/artificial feeding months 4.4%, p < 0.001. Our results indicate that there is not a simple and obvious relation between prandial liquid aspiration and pneumonia. Artificial feeding does not seem to be a satisfactory solution for preventing pneumonia in elderly prandial aspirators. PMID- 8721069 TI - Quantitative assessment of swallowing in healthy adults. AB - Swallowing has hitherto been evaluated during physical examination, radiologic barium studies, manometry, and cervical auscultation. Radiography principally demonstrates qualitative aspects of oral and pharyngeal function, whereas quantitative aspects have primarily been documented by manometry. To evaluate swallowing quantitatively, without using invasive methods or radiation, we have applied a combined test of water drinking, i.e., the Repetitive Oral Suction Swallow test (ROSS). The test provides reliable measurements of suction pressure, bolus volume, timing of important events in oral and pharyngeal swallow, and respiration. The test is described and results from 292 healthy, non-dysphagic subjects are presented. We found a mean bolus volume of 25.6 +/- 8.5 ml during single swallow and 21.1 +/- 8.2 ml during stress (forced, repetitive swallow). During forced, repetitive swallow, the bolus volume was more strongly associated with suction time (r2 = 0.55) than with peak suction pressure (r2 = 0.04), indicating that suction time is more important than suction pressure in determining the bolus volume. The oral-pharyngeal transit time decreased: single swallow 0.56 +/- 0.36 sec, forced repetitive swallow 0.23 +/- 0.11 sec, as did the coefficient of variation (48% and 64%, respectively) indicating a more automatic neural process for pharyngeal function in forced, repetitive swallow. The postswallow respiration started with inspiration in 10% of studied individuals, but did not correlate with deviations in other variables in the test. Thus, postswallow inspiration must be considered as normal. The ROSS test offers a rapid and easy quantitative assessment of swallowing. PMID- 8721070 TI - The influence of bolus volume and viscosity on anterior lingual force during the oral stage of swallowing. AB - The influence of bolus volume and viscosity on the distribution of anterior lingual force during the oral stage of swallowing was investigated using a new force transducer technology. The maximum force amplitudes from 5 normal adults were measured simultaneously at the mid-anterior, right, and left lateral tongue margins during 10 volitional swallows of 5-, 10-, and 20-ml volumes of water, applesauce, and pudding. Results indicated significant increases in peak force amplitude as viscosity increased. Volume did not significantly influence maximum lingual force amplitudes. Individual subjects demonstrated consistent patterns of asymmetrical force distribution across the lingual margins tested. The results suggest that bolus-specific properties influence the mechanics of oral stage lingual swallowing. This finding has important clinical implications in the assessment and treatment of dysphagic individuals. PMID- 8721071 TI - Unexplained dysphagia: viscous swallow-induced esophageal dysmotility. AB - Dysphagia is a manifestation of several clinical conditions of diverse origin. In spite of the variation in these disease entities in terms of their etiology, clinical presentation, natural history, and treatment, the mechanism of this clinical complaint is not always clear. We studied a group of patients with dysphagia for solids in whom no anatomic or motor abnormalities were encountered on standard studies. The group consisted of 37 patients, 25 women and 12 men, who were complaining of dysphagia of 6 months or longer duration and they did not demonstrate structural or motor abnormalities on barium esophagogram, esophagoscopy, and standard esophageal manometry. A group of 24 age-matched patients, 14 women and 10 men, with noncardiac chest pain served as the patient control. Esophageal contractile activities were studied after 10 wet swallows (5 ml of water) and 10 viscous swallows (5 cubic cm of marshmallow). Resting lower esophageal sphincter pressure and its relaxation response to swallows, amplitude of peristaltic activities, rate of dysphagia provoked during the study, and the frequency of abnormal esophageal contractions were evaluated. Six abnormal esophageal contractile activities-failed peristalsis, dropout, repetitive, simultaneous, spontaneous contractions, and aperistalsis-were utilized to generate an esophageal peristaltic dysfunction index. The mean LESP was 8.1 +/- 4.7 in the dysphagia group and 16.1 +/- 4.3 in the chest pain group. The mean amplitude of peristaltic contractions was 47.1 +/- 16.1 and 89.0 +/- 27.0 mmHg after wet swallows for dysphagia and chest pain groups, respectively. These values were 58.2 +/- 12.4 and 92.4 +/- 22.1 for viscous swallows. Swallowing provoked dysphagia in 89% of the dysphagia group after viscous swallows and 9% after wet swallows. In contrast, only 11% and 3% of control group complained of dysphagia during the study. This group of patients probably represent a cohort of patients with a nonspecific esophageal motor disorder in whom both clinical symptom and their esophageal motor counterpart can only be elicited in response to viscous swallows. We strongly believe in addition of viscous swallows in evaluating dysphagic patients in whom symptoms remain unexplained in light of standard studies. PMID- 8721072 TI - Management of oral-pharyngeal dysphagia symptoms in amyotrophic lateral sclerosis. AB - Oral and pharyngeal dysphagia is a common symptom in patients with amyotrophic lateral sclerosis (ALS) and is the result of a progressive loss of function in bulbar and respiratory muscles. Clinicians involved in the management of ALS patients should be familiar with the common clinical findings and the usual patterns of temporal progression. The prevention of secondary complications, such as nutritional deficiency and dehydration that compound the deteriorating effects of the disease, requires careful monitoring of each patient's functional status and timely intervention with appropriate management techniques. PMID- 8721073 TI - Swallowing in hereditary sensory ataxia. AB - The oral, pharyngeal, and esophageal stages of swallowing were evaluated in 8 patients with recessively or dominantly inherited pure sensory ataxia. Six patients had swallowing difficulties: solid bolus obstruction, coughs during eating, and choking episodes. One patient had chronic bronchitis and another had recurrent pneumonia. The patients underwent a biphasic radiological barium swallow, including videofluoroscopy. No patient had a completely normal swallow. All had normal oral function, whereas pharyngeal function was abnormal in 6 patients. Esophageal function was abnormal in 6 patients. The swallowing dysfunction did not correlate with the severity of motor or sensory dysfunction in the limbs, nor with age or duration of ataxia. Our study shows that swallowing dysfunction is common in hereditary sensory ataxia. This dysfunction is likely to be due to involvement of the nucleus of the solitary tract in the brainstem. Despite some of the patients having suffered from choking episodes and others from bronchopulmonary complications, they did not spontaneously admit dysphagia. Swallowing should be evaluated thoroughly in patients with hereditary sensory ataxia since dysphagia in these patients might bring serious and potentially fatal complications. PMID- 8721074 TI - Quantitative assessment of oral and pharyngeal function in Parkinson's disease. AB - Oral and pharyngeal dysfunction is common in Parkinson's disease. To reveal the frequency of swallowing dysfunction and correlate swallowing dysfunction with locomotor disturbances, we studied 75 patients with Parkinson's disease staged I IV according to the Hoehn and Yahr score. We assessed oral and pharyngeal swallow during optimal medication by a quantitative test of swallowing (the ROSS test) measuring the suction pressure, bolus volume, swallowing capacity, and time for important events in the swallowing cycle. We found abnormal results in 7/12 patients (58%) in stage 1 of the Hoehn and Yahr score, in 13/14 patients (93%) in stage 2, in 29/32 patients (91%) in stage 3, and in 16/17 patients (94%) in stage 4. Abnormal test results in stages, 1, 2, and 3 were seldom related to swallowing difficulties noticed by the patients. In advanced disease (Hoehn and Yahr stage 4), the abnormal results were often considerable, with swallowing difficulties obvious to the patient. Two of 17 patients coughed during or immediately after the test and 3/ 17 patients were unable to complete the test. The degree of swallowing disturbance increased during stress (forced, repetitive swallow). The Hoehn and Yahr score and the results in the ROSS test did not correlate, indicating that swallowing disturbances are due to nondopaminergic degeneration. Silent swallowing impairment may interfere with the nutrition and quality of life in Parkinson's disease, thus it is of interest to monitor this in clinical practice. PMID- 8721075 TI - Dysphagia in drug-induced parkinsonism: a case report. AB - Dysphagia complicates both idiopathic Parkinson's disease (IPD) and drug-induced parkinsonism (DIP). Although parkinsonism of DIP and IPD are often clinically indistinguishable, there is no assurance that their abnormalities of swallowing will be similar. We evaluated a patient with DIP who complained of difficulty chewing and swallow initiation. The dysphagia evaluation demonstrated abnormalities during all stages of ingestion. However, the prepharyngeal stages were disproportionately affected when compared with patients with IPD and similar levels of parkinsonian functional disability. This case gives additional support for a significant basal ganglia influence on motor deglutitive functions. PMID- 8721076 TI - [Susceptibilities of bacteria isolated from patients with respiratory infectious diseases to antibiotics (1993)]. AB - Bacteria isolated from respiratory tract infections were collected in cooperation with institutions located throughout Japan, since 1981, and the Ikemotor et al. have been investigating susceptibilities of the isolates of various antibacterial agents and antibiotics, and the relationships between the isolates and backgrounds of the patients and so forth each year. We discuss the results in detail. In 20 institutions around the entire Japan from October 1993 to September 1994, 584 strains of bacteria were isolated mainly from sputa of 473 patients with respiratory tract infections and presumed to be the etiological agents. MICs of various antibacterial agents and antibiotics were determined against 91 strains of Staphylococcus aureus, 98 strains of Streptococcus pneumoniae, 122 strains of Haemophilus influenzae, 91 strains of Pseudomonas aeruginosa (non mucoid), 34 strains of Pseudomonas aeruginosa (mucoid), 42 strains of Moraxella subgenus Branhamella catarrhalis, 25 strains of Klebsiella pneumoniae etc., and the drug susceptibilities of these strains were measured except the strains which died during transportation. 1. S. aureus S. aureus strain sfor which MICs of methicillin was higher than 4 micrograms/ml (methicillin-resistant S. aureus) accounted for 56.0%, but this frequency of the drug resistant bacteria was lower than the previous year's 61.4%. Arbekacin and vancomycin showed the highest activities against MRSA and MIC80s were 1 microgram/ml. 2. S. pneumoniae Benzylpenicillin among the penicillins showed potent activities against S. pneumoniae. Cefuzonam, cefotaxime and cefmenoxime among the cephems showed excellent antimicrobial activities against S. pneumoniae. Imipenem; carbapenems, showed the most potent activity, and MIC90 was 0.063 microgram/ml. 3. H. influenzae All the drugs tested were quite active against H. influenzae. Cefotaxime, cefmenoxime, cefuzonam and cefixime among the cephems showed the most potent activities, and MIC90 were 0.063 microgram/ml against H. influenzae. Ofloxacin also showed MIC90 of 0.063 microgram/ml. 4. P. aeruginosa (mucoid) Tobramycin showed the most potent activity against P. aeruginosa (mucoid), and MIC80 was 1 microgram/ml. Ceftazidime, cefsulodin, imipenem, aztreonam, gentamicin and ciprofloxacin showed potent activities with MIC80s of 2 micrograms/ml. 5. P. aeruginosa (non-mucoid) Tobramycin showed the highest activity against P. aeruginosa (non-mucoid), and MIC80 was 1 microgram/ml, followed by ciprofloxacin with MIC80 of 2 micrograms/ml. Comparing to activities against P. aeruginosa (mucoid), all the drugs tested had relatively low activities against P. aeruginosa (non-mucoid). 6. K. pneumoniae. The activities of all drugs except ampicillin and minocycline were high against K. pneumoniae. Cefozopran, imipenem and carumonam showed the highest activities and MIC80s were 0.125 microgram/ml. Flomoxef showed the next highest activities with an MIC80 of 0.25 microgram/ml. 7. M.(B.) catarrhalis Imipenem showed the most potent activity against M.(B.) catarrhalis, with an MIC80 of 0.063 microgram/ml, followed minocycline and ofloxacin with their MIC80s of 0.125 microgram/ml. We also investigated year to year changes in the background of patients, as well as types of respiratory infectious diseases, and the etiological agents. As for patients background, there were many infectious diseases found among patients a high age bracket, and the patients over age 60 accounted for 61.3% of the diseases. The distribution by respiratory tract infections was as follows: chronic bronchitis and bacterial pneumonia accounted for the greatest numbers of cases with 31.1% and 26.0%, respectively, followed by bronchiectasis with 10.4%. In this year chronic bronchitis under age 29 were 41.7%, thus was much higher than 12.5% in previous year. This marked change was first noted in your research during the recent 5 years. As for frequencies of etiologic bacteria by respiratory tract infections, S. pneumoniae (ABSTRACT TRUNCATED) PMID- 8721077 TI - [A comparative study on the efficacies of ritipenem acoxil and cefotiam hexetil in bacterial pneumonia by the double-blind method]. AB - To objectively evaluate the efficacy, safety and usefulness of the newly developed penem oral antibiotic, ritipenem acoxil (RIPM-AC), against bacterial pneumonia, we conducted a multi-center double-blind comparative study using cefotiam hexetil (CTM-HE) as the control drug. Both RIPM-AC and CTM-HE were orally administered at 200 mg t.i.d. for 14 days, in principle. The results were as follows: The total number of patients enrolled in this trial was 208, of which 152 cases (RIPM-AC group: 73, CTM-HE group: 79) were evaluable for clinical efficacy. 1. The clinical efficacy rates (excellent + good) were 91.8% (67/73) in the RIPM-AC group and 94.9% (75/79) in the CMT-HE group. There was no significant difference between the two groups, and the clinical equivalency of RIPM-AC to CTM HE was demonstrated. 2. In the patients enrolled in the evaluation of clinical efficacy, the eradication rates of the causative organisms were 84.6% (22/26) in the RIPM-AC group and 91.7% (22/24) in the CTM-HE group, with no significant difference between the two groups. 3. Side effects were noted in 9 cases (9.6%) of the RIPM-AC group and 5 cases (4.9%) of the CTM-HE group. Abnormal laboratory test findings were observed in 23 cases (26.7%) of the RIPM-AC group and 15 cases (15.6%) of the CTM-HE group. There was no significant differences between the two groups in the incidence of side effects nor of abnormal laboratory test findings. In the safety evaluation, RIPM-AC was judged to be safe in 64 cases (68.1%) and CTM-HE in 82 cases (80.4%), with no significant difference. 4. The usefulness rates (markedly useful+useful) were 86.5% (64/74) in the RIPM-AC group and 92.5% (74/80) in the CTM-HE group. There was no significant difference between the two groups. Since RIPM-AC showed clinical efficacy similar to those of CTM-HE and posed no particular safety problems, it is expected to be a useful antibiotic for the treatment of bacterial pneumonia. PMID- 8721078 TI - [Antimicrobial activities of meropenem against clinically isolated strains]. AB - In order to evaluate antimicrobial activity of meropenem (MEPM), minimum inhibitory concentrations (MICs) of MEPM and control drugs were determined against clinical isolates in 1993. The results were as follows; 1. Antimicrobial activities of MEPM against Gram-positive bacteria were stronger than those of cephems (CEPs), were approximately equal to those of panipenem (PAPM), and were weaker than those of imipenem (IPM). 2. Carbapenems showed strong antimicrobial activities against Enterobacteriaccae, glucose non-fermentative Gram-negative rods and Bacteroides fragilis group that were multiple drug resistant including the third generation CEPs. Antimicrobial activities of MEPM against these organisms were stronger than those of IPM and PAPM. 3. MIC-ranges of MEPM against Enterobacteriaceae and Haemophilus influenzae were lower than those of IPM and PAPM. We observed that MEPM had better permeability into the cells of H. influenzae, higher affinities to 3 to 5 different penicillin-binding protein and high stability against beta-lactamase than those of IPM and PAPM. PMID- 8721079 TI - [Antibacterial activities of combination uses of isepamicin and beta-lactams in vitro against clinically isolated strains. Part 1. Activities against Staphylococcus aureus]. AB - In order to evaluate antibacterial activities of combination uses of isepamicin (ISP) and beta-lactams in vitro, minimum inhibitory concentrations (MICs) these drugs were examined singly and in combination against clinically isolated Staphylococcus aureus. The results are summarized as follows; 1. MICs of ISP + cefazolin (CEZ), ISP + cefotiam (CTM) and ISP + flomoxef (FMOX) were low and the activities against methicillin (DMPPC)-susceptible S. aureus (MSSA) were dependent on the concentration of ISP. Combined effects were observed when the concentrations of ISP were at sub-MIC levels (1/2 approximately 1/4 concentrations). 2. MICs of ISP + CEX, ISP + CTM, ISP + FMOX, ISP + imipenem and ISP + panipenem were low and the activities against DMPPC-resistant S. aureus (MRSA) were dependent on the concentration of ISP, and were similar to those against MSSA. Combined effects were observed when the concentrations of ISP were at sub-MIC levels of ISP. Lower MIC50 or MIC90 was observed at ISP concentrations of 4 approximately 16 micrograms/ml. 3. The blood Cmax of ISP exceeded 20 micrograms/ml at one-time administration of ISP 400 mg, and these results suggested that antibacterial activities of combination uses of ISP and beta lactams was clinically effective against MRSA infections. PMID- 8721080 TI - [Study of the usefulness of a kit containing imipenem/cilastatin powder with diluent for injection: convenience of use and preparation]. AB - The usefulness of a newly developed imipenem/cilastatin powder-and-diluent kit packed in non-glass container was compared to that packed in commercially available glass vials. The imipenem/cilastatin powder-and-diluent kit container is made of a polyolefin bag with two chambers that contain imipenem/cilastatin powder and diluent (0.9% saline, 100 ml), respectively. The convenience of use of the kit and the time required for the preparation of the dosing solution were evaluated by nurses and pharmacists at the Kitasato University East Hospital. The newly developed kit received higher scores with regard to convenience of use and led to a 46 approximately 59% reduction in the time required for preparation as compared to the commercially available glass vials. PMID- 8721081 TI - [Study of the usefulness of a kit containing imipenem/cilastatin powder with diluent for injection: accuracy of reconstitution]. AB - The usefulness of a kit consisting of non-glass packaging was evaluated in terms of accuracy of reconstitution. The newly developed imipenem/cilastatin powder-and diluent kit consists of a polyolefin bag with two chambers that contain impenem/cilastatin powder and diluent (0.9% saline, 100 ml), respectively. The accuracy of reconstitution was determined by nurse and pharmacists at the Kitasato University East Hospital by measuring the amount of materials remaining in vials after reconstitution using the syringe dilution method and the transfer needle dilution method. The mean percent amounts of imipenem and cilastatin remaining after preparation by the syringe dilution method were 5.58 +/- 2.60% and 4.08 +/- 1.77%, respectively. The mean percent amounts of imipenem and cilastatin remaining after preparation by the transfer-needle dilution method were 3.99 +/- 2.28% and 3.71 +/- 2.09%, respectively. The amount of imipenem/cilastatin remaining in the newly developed kit should be negligible, because the kit serves as both a vial and a dosing package. Therefore, greater accuracy in terms of reconstitution is expected with the newly developed kit than with the traditional syringe dilution or transfer-needle dilution methods. PMID- 8721082 TI - In vitro drug response assays for entry into the rational era cancer chemotherapy. PMID- 8721083 TI - [Tumor-rejection antigens expressed on human squamous cell carcinoma]. AB - Squamous cell carcinoma (SCC) is one of the most common cancers in human. SCC, particularly, esophageal and lung SCC are relatively resistant to currently available regimens of chemotherapy or radiation therapy. Therefore, development of a specific immunotherapy using tumor specific cytotoxic T lymphocytes (CTL) would be important to offer other treatment modalities. However, generation of HLA class I-restricted CTL recognizing SCC has been rarely reported. We established the HLA A2601-restricted CTL cell line recognizing a peptide antigen expressed on SCC. This CD4- CD8+ cytotoxic T lymphocyte (KE-4 CTL) cell line was established in a patient with esophageal cancer. The KE-4 CTL recognized a peptide antigen on esohageal and lung SCC in an HLA A2601-restricted manner as evaluated by cytotoxity against a panel of tumor cells, transfection experiments with HLA A2601 cDNA, and reconstitution with eleted peptides. None of normal cells tested was lysed by this CTL. These results suggest the exstence of HLA A2601-restricted CTL precursors recognizing a peptide antigen on SCC in a patient with esophageal cancer. PMID- 8721084 TI - [Chemosensitivity of cultured meningiomas]. AB - Meningioma is one of the popular benign brain tumors. However, the recurrence of this tumor is not infrequently encountered. In an attempt to establish the useful adjuvant therapy for the recurrent meningioma, in vitro chemosensitivity study for meningioma was conducted. Among various chemotherapeutic agents tested here, cisplatin showed highest cytotoxicity on cultured meningioma cells. In conclusion, cisplatin may be useful in adjuvant chemotherapy for the recurrent meningioma. PMID- 8721085 TI - [Biological features determining the chemosensitivity of gastric cancer]. AB - We analysed the relationship between several biological properties of gastric cancers and their chemosensitivity determined by MTT assay. Higher chemosensitivity was associated with poor differentiation, aneuploidy, and higher proliferative activity. Lymph node metastasis was more chemosensitive than primary lesion, while liver metastasis was less. Gastric cancer expressing multidrug-resistance associated protein (MRP) showed lower sensitivity to several anticancer drugs, including adriamycin and etoposide. p53 status and susceptibility to apoptosis were also associated with chemosensitivity. Thus, chemosensitivity of clinical gastric cancer might be increased if these characters can be modified by some new biologic therapy. PMID- 8721086 TI - [Evaluation of drug sensitivities to gastric cancers by SDI test--study of quality control of SDI test by simulation test]. AB - For the purpose to evaluate the feasibility of SDI test, a drug sensitivity test, against resected gastric cancer, a multiinstitutional study consisted of multiple institute, the 16th specified study was planned by Japanese Foundation for Multidisciplinary Treatment of Cancer. SDI test was performed individually in 15 institutes in Japan using surgically resected specimens from the patients with gastric cancer. The strict quality control was necessary to unify the method and the technique of this test in 15 institutes. Tumor bearing nude mice were delivered to each institute and the test was performed simultaneously. The data of test were sent by personal computer to Department of Surgery II Nagoya University. These data were estimated by two factors. One is the deviation of data in one institute and the other is the deviation of data among 15 institutes. This simulation test was continued through this study, 4 institutes could not pass this simulation test but the data from other three institute indicated excellent uniformity. PMID- 8721087 TI - [Potential and limitation of scintillation assay (TIA) for clinical chemotherapy]. AB - The clinical significance of the scintillation assay (thymidine incorporation assay) which we developed was summarized as follows; 1)thymidine uptake by tumor cells which can be evaluated in each assay functioned as a significant predictor of prognosis of patients with gastric or colorectal cancer, 2)the tumor cell compartment in this assay system significantly increased more than did those in other culture systems, 3)evaluable rates were increased to more than 80% by introduction of the preculture system with collagen coated flasks, and 4)it can be considered that chemosensitivity testings may benefit for some types of human tumor, but not for all. PMID- 8721088 TI - [Studies on the in vivo and in vitro chemosensitivity tests using human tumor xenografts]. AB - The in vitro chemosensitivity test has been appreciated as an useful laboratory method for selecting the optimal anticancer drug for solid carcinomas. Using human tumor xenografts, the correlation of in vitro and in vivo tests was studied. The sensitivity results were identical between the two methods for 75% of samples tested. In the other study, five surgically resected specimens were subjected to the in vitro test and were transplanted into nude mice. The in vitro tests were repeated on the xenografts, demonstrating satisfactory reproducibility of the sensitivity results. PMID- 8721090 TI - [Cancer chemosensitivity test--from laboratory to clinic]. AB - The endpoint of chemosensitivity test lies to extend the survivals of cancer patients using the adapted antitumor agents determined by the assay. Although the overall predictive accuracy of chemosensitivity test was 79% (117/149) for gastrointestinal carcinomas, no remarkable survival benedit was obtained even in the "true positive" cases. Two retrospective and one semi-prospective studies were conducted using MTT assay or histoculture drug response assay, indicating that these tests will be useful in evaluating the adoptive adjuvant cancer chemotherapy for advanced gastric cancer by increasing the patients' survivals. The chemosensitivity test should be changed its position from laboratory to clinic through an acceptance of the assay as social medical insurance. PMID- 8721089 TI - [Effects of chemotherapy on the basis of the results of MTT assay for patients with gastrointestinal cancer]. AB - The chemosensitivity test should be performed to individualize the chemotherapy for patients with gastrointestinal cancer, which is one of the tumors most refractory to treatment by anticancer drugs. The present study was designed to determine the chemosensitivity in fresh human gastrointestinal cancer, using highly purified tumor cells, and the correlation of this sensitivity with clinical response. The clinical responses were obtained in 15 of the 25 patients, and 5 of the 15 patients with gastric cancer and colorectal cancer, respectively. The inhibition rates for anticancer drugs in responders were higher than in nonresponders. Thus, it is suggested that the chemotherapy according to the results of the MTT assay is effective in patients with gastrointestinal cancer. PMID- 8721091 TI - [Establishment of a new human endometrial adenocarcinoma cell line, Watanabe cells, containing estrogen receptor]. AB - A new human endometrial adenocarcinoma cell line, Watanabe cells, was established from the ascitic fluid of a relapsed endometrial adenocarcinoma obtained from a 58-year-old woman; this cell line has been maintained in vitro for more than 3 years and 8 months. The cells formed a monolayer in a mosaic fashion and tended to pile up and formed a hemicyst. The population boubling time was 60.0 hours at the 10th generation. The modal chromosomal number of the cells was in the diploid range. The histology of tumors induced by this cell line in athymic nude mice showed poorly differentiated adenocarcinoma, while the initial tumor was a well differentiated adenocarcinoma. Estrogen and progesterone receptors (ER, PR) were demonstrated in the original tumor, whereas ER but not PR were present in the tumors induced in nude mice. CA125, CA19-9 and other tumor markers were positive in culture media of this cell line. The cells showed intrinsic cisplatin resistance (50% inhibition concentration: > 10 micrograms/ml) at 120 hours of exposure by MTT assay. We believe this cell line will be useful for investigating the mechanisms of progesterone therapy, the biological behaviors of the tumor markers and mechanisms of chemotherapeutic resistance in endometrial carcinoma. PMID- 8721092 TI - Gene transcripts of eleven proteins with specific functions are all detected in human normal cells and tumor cell lines: a possible DNA-->RNA basal constant flow. AB - Existance of messenger ribonucleic acids of eleven functionally differentiated proteins in normal human cells and tumor cell lines was investigated using reverse transcription--nested polymerase chain reaction method. Examined gene transcripts were those of progesterone receptor, estrogen receptor, interleukin 2, CD8, parathyroid hormone, cholecystokinin/pancreozymin, glucagon, insulin, adrenocorticotropic hormone, enkephalin and thyroid stimulating hormone. In RT PCR almost all primers were originally designed and sequences of PCR products were confirmed by the Sanger's method. Investigated cells were normal human peripheral mononuclear cells and their subsets, lymphokine-activated killer cells, gastric mucosal cells, sperm and fifteen established tumor cell lines. All of the examined mRNAs were detected in all of the above cell groups. Therefore, cells of independent tissues or tumors share same kinds of mRNA such as steroid hormone receptors, cytokine, lymphocyte surface molecule and parathyroid, digestive and cerebral hormones. These findings strongly suggest that every cell can express every mRNA. Beneath the cell differentiation there may exist a DNA- >RNA basal constant flow, the arrow of time captured in a cell. PMID- 8721093 TI - [Simplified method of cultured brain microcapillary endothelial cells]. AB - Many kinds of methods cultured brain microcapillary endothelial cells have been published, but these methods were not established. We designed the simplified methods with improvement of enzymatic treatment and Percoll gradient centrifugation. PMID- 8721094 TI - [Cytokines and growth of cancers]. PMID- 8721095 TI - [An experimental study on the physical examination system for psychic in patients. 2nd report: a trial screening for stomach and uterine cancer]. PMID- 8721096 TI - [Investigation of the actual conditions of hospital nurses working on three rotating shifts: questionnaire results of shift work schedules, feelings of sleep and fatigue, and depression]. AB - These studies were performed to clarify (1) the actual conditions concerning rotating shift schedules of nurses in Japanese university and college hospitals and to evaluate (2) some aspects of the physical and mental health, and (3) sleep profile of hospital nurses working on counter-clockwise shift rotation. Two questionnaire surveys and the OSA sleep inventory (OSA) were carried out. The subjects in the study (1) were a total of 80 nursing directors in university and college hospitals. The questionnaire covered 4 categories, such as the schedule most frequently adopted and reasons for using the schedule. The questionnaires were returned by 67 directors (83.8%). The subjects in the study (2) were 189 nurses working on three-shift work schedules at Asahikawa Medical College Hospital. The items in the questionnaire covered 7 categories, as follows: 1) feeling of sleep after each shift (8 items); 2) feeling of fatigue after each shift (30 items); 3) physical symptoms; 4) inter-personal problems; 5) all the items on Zung's self-rating depression scale (SDS); 6) all the items on the Horne and Ostberg morningness-eveningness questionnaire; and 7) 24 items on the Maudsley personality inventory. The questionnaires were returned by 156 nurses (82.5%), whose mean age and duration of shift-work employment were 27.2 +/- 5.1 and 5.0 +/- 4.3 years (mean +/- SD), respectively. For 152 nurses (97.4%) of those returning the questionnaire, the working schedule consisted of 2 consecutive night shifts and 2 consecutive evening shifts, following a variable number of day shifts (rapid and counterclockwise shift rotation). The subjects in the study (3) were 8 healthy nurses working on above-mentioned three rotating shifts at the psychiatric ward of Asahikawa Medical College Hospital, whose mean age was 29.4 +/- 5.8 years (mean +/- SD). All the subjects recorded their sleep logs and underwent OSA everyday for 30 consecutive days. Of the 240 OSA data, 95 data (16 after day shift, 17 after the 1st night shift, 16 after the 2nd night shift, 15 after the 1st evening shift, 16 after the 2nd evening shift, 15 after day off) were analyzed. In addition to five of sleep factors in the OSA analysis, we evaluated the global score (GS), which represents subjective global feeling of sleep. In the study (1), 47 of 66 hospitals (71.2%) adopted rapid and counterclockwise shift rotation. The results of study (2) were as follows: 1) After the first night shift (diurnal sleep), the sleep problems were worst, and the frequency of taking sleep-inducing drugs was highest (12.6%); 2) Feelings of fatigue were the highest level after each of the two night shifts; 3) SDS score was relatively high (57.8 +/- 8.1, mean +/- SD); 4) The older the nurse, the greater the aggravation of both sleep problems and fatigue; 5) Sleep problems after day shifts were worse and SDS score was higher in nurses classified as "nightowls" compared to those in nurses classified as "morning people"; and 6) There was no difference between introverts and extroverts in sleep problems, fatigue or SDS score. The results of study (3) were as follows: 1) In 3 of sleep factors (Sleepiness, Integrated sleep, sleep initiation) and GS, there were significant differences among each rotating shift schedule including day off; 2) The highest score in five of sleep factors and the highest GS were noted after the 2nd night shift (nocturnal sleep), and the lowest after the 1st night shift (diurnal sleep); and 3) The younger subjects (n = 4; mean age, 24.5 years) showed a higher GS compared with that of the older subjects (n = 4; mean age, 34.2 years). PMID- 8721097 TI - [The Italian situation in psychiatry after fifteen years of the psychiatric reform: the mental health service in north Italy]. PMID- 8721098 TI - [Basic study on relationship between estimated rate constants and noise in FDG kinetic analysis]. AB - For accurate estimation of the rate constants in 18F-FDG dynamic study, the shape of the estimation function (phi) is crucial. In this investigation, the relationship between the noise level in tissue time activity curve and the shape of the least squared estimation function which is the sum of squarred error between a function of model parameters and a measured data is calculated in 3 parameter model of 18F-FDG. In the first simulation, by using actual plasma time activity curve, the true tissue curve was generated from known sets of rate constants ranging 0.05 < or = k1 < or = 0.15, 0.1 < or = k2 < or = 0.2 and 0.01 < or = k3 < or = 0.1 in 0.01 step. This procedure was repeated under various noise levels in the tissue time activity curve from 1 to 8% of the maximum value in the tissue activity. In the second simulation, plasma and tissue time activity curves from clinical 18F-FDG dynamic study were used to calculate the phi. Results are: (1) In the noise-free case, because the global minima is separated from neighboring local minimums, it was easy to find out the optimum point. However, with increasing noise level, the optimum point was buried in many neighboring local minima. Making it difficult to find out the optimum point. (2) The optimum point was found within 20% of the convergence point by standard non-linear optimization method. (3) The shape of phi for the clinical data was similar to that with the noise level of 3 or 5% in the first simulation. Therefore direct search within the area extending 20% from the result of usual non-linear curve fitting procedure is recommended for accurate estimation of the constants. PMID- 8721099 TI - [Basic studies on the 111In labeled antisense oligonucleotide for tumor imaging]. AB - Antisense oligonucleotide labeled with short lived radionuclide has been proposed as a radiopharmaceutical for the detection of abnormal gene expression. In this study, plausibility of 32P or 111In labeled oligonucleotide was basically evaluated using c-erbB-2 protooncogene mRNA as a model target. In cell uptake studies, sequence specific accumulation of 32P-oligonucleotide was found in the cells with c-erbB-2 mRNA expression, but not control cells, indicating the basic possibility of antisense strategy. Synthesis of 111In labeled isothiocyanobenzyl EDTA (IBE)-oligonucleotide could be performed, but some problems were found in purification step. Stability of 111In-IBE-oligonucleotide was high when compared with that of 32P-oligonucleotide, and in vivo biodistribution data might indicate that 111In-IBE-oligonucleotide was plausible for tumor imaging. PMID- 8721100 TI - [Changes of regional perfusion in metastatic brain tumor and peritumoral area after radiosurgery: a study by 123I-IMP dynamic SPECT]. AB - Changes of regional perfusion in the tumor, peritumoral edematous area and juxtatumor brain after radiosurgical treatment for metastatic brain tumor were investigated by dynamic SPECT using 123I-IMP. The SPECT was performed in 12 patients before and 1, 7 and 30 days after stereotactic irradiation. A region of interest (ROI) was selected each in the tumor, peritumoral edematous area, juxtatumor brain and ipsilateral cerebellum. Radioactivity in each ROIs was counted on early SPECT based on dynamic SPECT from 0 to 5 minutes. Mean count/pixel in each ROIs was divited by mean count/pixel in the ROI of the ipsilateral cerebellum and its value was designated as a count ratio (CR). Assuming the pre-treatment CRs are 1.0, relative changes of post-treatment CRs investigated. rCR in the tumor did not show any significant change after radiosurgical treatment. rCR in the edematous area and the juxtatumor brain increased at 7 days after irradiation [Mean +/- SD 1.43 +/- 0.409 (p < 0.05), 1.248 +/- 0.228 (p < 0.05) by Mann-Whitney test] and at 30 days [1.359 +/- 0.245 (p < 0.01), 1.301 +/- 0.287 (p < 0.01)] respectively. Computed tomography revealed no change in the maximum diameter of the tumor at 1 month after irradiation but a significant reduction in the diameter [0.744 +/- 0.227 (p < 0.02)] at 2 months. Early improvement of regional cerebral blood flow in the juxtatumor areas after radiosurgery suggested that radiosurgery could be effective treatment for metastatic brain tumor. PMID- 8721101 TI - [The graft flow and 201Tl-myocardial scintigram in patients with saphenous vein graft or left internal thoracic artery graft]. AB - To evaluate myocardial perfusion in patients with saphenous vein graft (SVG) or internal thoracic artery graft (ITA-G), we studied 38 patients (14: SVG, 10 males and 4 females, mean age 66 +/- 9 y-o; 24: ITA-G, 18 males and 6 females, mean age 64 +/- 7 y-o) by digital subtraction angiography (DSA) of ITA-G or SVG, and thallium-201 myocardial perfusion scintigraphy on exercise or dipyridamole stress. The grafting sites were left anterior descending artery (LAD) in all patients. Normal controls (n = 22) were defined by normal coronary angiogram and no evidence of myocardial ischemia. The graft flow and flow reserve on dipyridamole were measured by Rutishauser's formula. The basal blood flow of native normal ITA, SVG and ITA-G were respectively 72 +/- 24 ml/min, 51 +/- 23 ml/min, and 36 +/- 20 ml/min. The basal ITA-G flow was significantly lower than SVG-flow (p < 0.05). The flow reserves of SVG and ITA-G were respectively 2.32 +/ 0.65 and 1.78 +/- 0.59 (p < 0.02). The incidence of moderate hypoperfusion of thallium-201 SPECT was 14.3% in SVG and 12.5% in ITA-G on exercise stress, and 35% in SVG and 50% in ITA-G on dipyridamole stress. The incidence of reversible myocardial ischemia on dipyridamole stress was significant. The graft flow in patients with normal and abnormal thallium-201 SPECT were respectively 61 +/- 21 ml/min and 33 +/- 15 ml/min in SVG (p < 0.01), 46 +/- 19 ml/min and 27 +/- 16 ml/min in ITA-G (p < 0.02). The graft flow reserve were respectively 2.69 +/- 0.38 and 1.65 +/- 0.49 in SVG (p < 0.001), 2.25 +/- 0.40 and 1.31 +/- 0.28 in ITA G (p < 0.001). We concluded that the basal blood flow and flow reserve of ITA-G were significantly lower than those of SVG. The myocardial ischemia was occasionally documented by the thallium-201 myocardial SPECT on dipyridamole stress in patients with patent ITA-G. PMID- 8721102 TI - [Effects of compton scatter in quantitative brain SPECT]. AB - Effects of Compton scatter has been investigated for quantitative single-photon emission computed tomography. Using a technique proposed by Ichihara and Ogawa (triple-energy window method) as a gold standard, effects of correcting the scatter has been evaluated for 5 different phantom configurations, as well as a clinical cerebral blood flow study with use of 123I-IMP. Without the scatter correction but with a correction for attenuation, a conventional reconstruction provided a non-uniform distribution for uniform phantom configurations, and the quantitative pixel counts being highly dependent on size and shape of the objects. On the other hand, a uniform distribution and quantitative pixel counts that were independent of size or shape of the object were obtained by applying the scatter correction using a conventional attenuation correction technique with use of a theoretical mu value of 0.146 cm-1, thus suggesting importance of the scatter correction. Similar results were observed without the scatter correction, if an empirical value of mu of 0.07 cm-1 was used in the attenuation correction. However, this procedure was found to decrease the image contrast between the high count and low count regions. Significant underestimation of 20-30% was caused in cortical gray matter regions, and overestimation of 20% in the white matter regions. In practical clinical studies, magnitude of the error is still unknown, and is probably dependent on shape, size and radioactivity distribution of the object. A further systematic study is required in order to investigate significance of the scatter correction in real clinical studies. PMID- 8721103 TI - [99mTc-MIBI myocardial tomography with intravenous infusion of adenosine triphosphate in the diagnosis of coronary artery disease]. AB - To evaluate its feasibility, safety and diagnostic accuracy, 99mTc-MIBI myocardial tomography with adenosine triphosphate (ATP) infusion (0.16 mg/kg/min for 5 min) was performed 100 consecutive patients using the stress/rest one day protocol. None of the patients required treatment with aminophylline during the study. The sensitivity and specificity for detecting patients with coronary artery disease were 97% and 71%, respectively. Those for detecting individual coronary lesion (> or = 75% stenosis) were 92% and 89%, respectively. The high hepatic uptake of 99mTc-MIBI causes artifactual perfusion defects in the inferior myocardial wall, particularly on ATP stress images. In order to reduce this artifactual phenomenon, the interval time between injection and stress imaging must be increased. PMID- 8721104 TI - [Evaluation of reserved hepatic function in patients with hepatobiliary tumor by 99mTc-GSA: effect of hyperbilirubinemia and usefulness of regional reserved hepatic functional imaging]. AB - The evaluation of the reserved hepatic function was performed by 99mTc-galactosyl serum albumin (99mTc-GSA) in seventy patients with hepatobiliary tumor. The dynamic study was performed to evaluate global reserved hepatic function following the intravenous bolus injection of 99mTc-GSA, and the hepatic single photon emission computed tomography (SPECT) was obtained to assess the regional reserved hepatic function. The functional hepatic index (LHL15) was derived from liver time-activity data, and it was compared with serum total-bilirubin level, serum albumin level and plasma disappearance rate of indocyanine green (ICG15). In the patients with hepatocellular carcinoma, LHL15 value agreed well with ICG15 value, serum total-bilirubin level, and serum albumin level. Moderate or severe hepatic dysfunction was observed at 65.4% of these patients. In the patients with cholangiocellular carcinoma, a discrepancy of LHL15 value and ICG15 value was observed. Increment of the ICG15 value was correlated with that of the serum total-bilirubin level, whereas the correlations was not observed between the LHL15 value and the serum total-bilirubin level. These results indicate that 99mTc-GSA scintigraphy can evaluate the reserved hepatic function without the embellishment of jaundice. This method is useful for assessing the global and regional reserved hepatic function. PMID- 8721105 TI - [Anomalous origin of the left coronary artery from the pulmonary trunk with myocardial infarction and severe left ventricular dysfunction in infancy- assessment of myocardial damage using SPECT studies with 201TlCl and 123I-BMIPP]. AB - A 4-month-old male infant with Bland-White-Garland (BWG) syndrome complicated myocardial infarction was reported. Signs included tachypnea, coughing, and failure to thrive. However, there was no sign of myocardial infarction. A chest radiograph revealed cardiomegaly (CTR = 65%) and electrocardiogram showed abnormal Q waves in I, aVL, V6 leads. Cardiac catheterization and angiography revealed marked dilatation of left ventricle (end-diastolic volume = 384 ml/m2) and extremely depressed ejection fraction (16%), confirming the diagnosis of BWG syndrome. A 201TlCl-myocardial SPECT demonstrated apical defect and hypoperfusion in the anterolateral, inferoposterior walls, whereas 123I-beta-methyl-p iodophenylpentadecanoic-acid (123I-BMIPP) SPECT showed a wider defect area. SPECT studies with 201TlCl and 123I-BMIPP, are useful to assess myocardial viability more accurately in BWG syndrome. PMID- 8721106 TI - [Normal CBF values by the ARG method using IMP SPECT: comparison with a conventional microsphere model method]. AB - N-isopropyl-p[123I]iodoamphetamine (IMP) has been used as a flow tracer for SPECT, and measurement of cerebral blood flow (CBF) using IMP has been performed by conventional microsphere model method (MS method). Recently, the ARG method for measuring CBF by using IMP with one SPECT scan and one point blood sampling has been developed. This method was based on two-compartment model. In the present study, normal CBF values were measured in ten male healthy subjects (mean age +/- S.D.: 29.8 +/- 6.01, age range: 23-41) by the ARG and the MS methods. The mean CBF values (+/- S.D.) for the ARG method in which the Vd value was assumed to be 50 ml/ml were 41.7 +/- 9.4, 31.1 +/- 5.0, 40.7 +/- 9.7, 41.5 +/- 10.0, 38.2 +/- 9.2, 39.0 +/- 9.4, 41.9 +/- 10.6, 38.7 +/- 8.0 and 30.0 +/- 7.7 ml/100 ml/min in the cerebellum, pons, thalamus, basal ganglia, frontal, temporal, parietal, occipital lobe cortex and centrum semiovale, respectively. The mean CBF values for the MS method were 46.8 +/- 8.4, 37.5 +/- 5.6, 45.8 +/- 8.6, 46.5 +/- 8.9, 43.7 +/- 8.3, 44.4 +/- 8.7, 46.8 +/- 9.3, 44.3 +/- 7.3 and 36.3 +/- 8.1 ml/100 ml/min, respectively. The mean CBF values in the cerebral cortex region for the ARG method were lower than those previously reported by PET. This would be caused by low first-pass extraction fraction of IMP compared with oxygen-15 labeled water. The mean CBF values for the MS method were higher than those for the ARG method against previous studies. As reasons for this, errors in estimation of the SPECT brain counts at 8 min in the MS method were considered. PMID- 8721107 TI - [Phase 2 clinical study of 123I-iomazenil in various cerebral diseases: part 1- examination of injection dose and clinical indications]. AB - A phase 2 study of 123I-Iomazenil (IMZ) was performed to evaluate its clinical usefulness in SPECT imaging of central-type benzodiazepine receptors (BZR). A total of 246 patients with various central nervous system disorders [82 with epilepsy, 94 with cerebrovascular disease (CVD), 47 with degenerative disorders, 14 with mental disorders relating to BZR and 9 with other diseases] were intravenously injected with 111-222 MBq of IMZ. Early and late images were obtained 15 min and 180 min after injection as the mid-scan time, respectively. In comparison with regional cerebral blood flow (rCBF) images, the uptake of IMZ in the late images, which reflect the regional distribution of BZR, was relatively preserved compared to the uptake of CBF tracers in most of the patients with CVD. In the area of hyperperfusion in the subacute phase of cerebral infarction, however, the defects of IMZ uptake were accompanied by increased uptake of the CBF tracers. On the other hand, late IMZ uptake in most of the patients with the other diseases was decreased more significantly than the uptake of the CBF tracers, suggesting that cortical neuronal damage is more prominent than rCBF in these diseases compared to the vascular perfusional state. No adverse reactions were recognized relating to the IMZ administration. These results suggest that IMZ may provide us with valuable information for assessing pathophysiological state in the brain, which are difficult to achieve with other imaging modalities. PMID- 8721108 TI - [Phase 2 clinical study of 123I-iomazenil in various cerebral diseases: part 2- clinical evaluation of central-type benzodiazepine receptor imaging with 123I iomazenil SPECT]. AB - A phase 2 clinical study of 123I-iomazenil (IMZ) was performed to evaluate its clinical usefulness as central-type benzodiazepine receptor imaging agent in 81 cases with epilepsy, 82 cases with cerebrovascular diseases (CVD), 35 cases with degenerative disorders and 14 cases with mental disorders. In epilepsy, IMZ SPECT images for detection of epileptic foci showed relatively high specificity compared with EEG recording in not only interictal but also ictal state, in spite of the fact that IMZ SPECT images was performed during interictally. It was surmised that IMZ SPECT provides valuable clinical information for detection of epileptic foci which are hard to be diagnosed by EEG. The frequency of abnormal findings by IMZ late images was higher than that by interictal blood flow images and comparable to that by ictal blood flow images. In patients with CVD, different observations were obtained between IMZ late images and blood flow images; for example, in regions of hemodynamic cerebral ischemia, postischemic reperfusion and other pathological perfusion, IMZ late images seemed to reflect the degree of cortical neuronal loss. In patients with degenerative disorders, the binding potential (calculated by a simple quantitative method for IMZ) and other parameters correlated with the dementia score. While the number of examined cases with mental disorders was limited, the frequency of abnormal findings with IMZ imaging was higher than that with morphological imaging. PMID- 8721109 TI - [Cancer diagnosis with positron emission tomography (PET)]. AB - PET using 18F-fluorodeoxyglucose (FDG) or 11C-L-methionine (Met) enables cancer diagnosis based on higher metabolic demand of malignant tumor, that is correlated to uncontrolled cell proliferation. PET using FDG or Met is useful for differential diagnosis or staging of malignant tumor in brain, head and neck, lung, breast, adrenal, pancreas, and others. PET is also useful for detection of recurrent tumor in post-therapy scar or necrosis in colon, lung and brain. Recently, basic studies clarified the mechanisms of tracers accumulation in tumor tissue at the microscopic level, which will be helpful for the improvement of diagnostic accuracy of PET. PET can provide useful information to improve patient management in oncology. PMID- 8721110 TI - [Clinical analysis on the course and prognosis of patients with Crohn's disease]. AB - We analyzed 66 cases (47 males and 19 females) of Crohn's disease at Hiroshima University hospital from September 1975 to October 1994 to clarify the course and prognosis of Crohn's disease. The age at onset was 21.1 +/- 7.3 years old (mean +/- SD), terms between onset and diagnosis were 21.5 +/- 33.0 months (mean +/- SD) and observation period was 65.5 +/- 44.6 months (mean +/- SD). Sites of lesion were 18 ileum, 41 ileocolon and 7 colon. Thirty-one cases, 20 cases of which had intestinal obstruction, underwent surgical operation (12 ileum types, 18 ileocolic types, 1 colon type). The cumulative probability of surgery at one, five and ten years after onset of symptoms were 12.1%, 28.8% and 56.9%, respectively. As for cumulative probability of surgical operation at one, five and ten years after diagnosis were 25.8%, 36.7% and 74.4%, respectively. Results of the cumulative probability of surgery by anatomical involvement indicated that the ileum type had a statistically significantly higher risk than other types. In each analysis compliance to nutritional therapy was also an important prognostic factor. Overall, our results indicated that the site of lesion and the compliance to nutritional therapy were important factors which have an effect on the course and prognosis of Crohn's disease patients. PMID- 8721111 TI - [The role of catalase in histologic changes of acute experimental pancreatitis]. AB - We produced acute biliary reflux pancreatitis by an injection of auto-bile in 31 young mongrel dogs. We divided into control group (n = 10), catalase administration group (B1 group, n = 17) and no-administration group (B2 group, n = 14). We studied the histologic changes of the brain tissue in those groups. There was no significant difference in the survival rate between group B1 and B2. In the histologic findings, ischemic cell changes were dominant in both groups within one month, but after 3 or 6 months after operation in group B2, satellitosis and/or neuronophagia was recognized as the irreversible changes. The lipid peroxides level was significant high in group B2. Consequentially, it is natural to think that the production of free radicals have been inhibited by catalase and the histologic changes in the brain in group B1 have been light. PMID- 8721112 TI - [A case of symptomatic aberrant pancreas: a diagnostic and therapeutic usefulness of endoscopic aspiration mucosectomy]. PMID- 8721113 TI - [A case of ischemic colitis associated with interferon treatment]. PMID- 8721114 TI - [A case of melanosis duodeni]. PMID- 8721115 TI - [Hepatitis with jaundice associated with rubella infection in a patient with intestinal Behcet disease; report of a case]. PMID- 8721116 TI - [Two case of Fitz-Hugh-Curtis syndrome with Chlamydia trachomatis infection]. PMID- 8721117 TI - [A case report of congenital liver fibrosis tolerated 3 normal deliveries]. PMID- 8721118 TI - [A case of combined or mixed hepatocellular and cholangiocarcinoma with sarcomatous components--immunohistochemical characteristics and histogenesis]. PMID- 8721119 TI - [A case of autoimmune cholangiopathy which developed 11 years after occurrence of SLE]. PMID- 8721120 TI - [A case of tiopronin-induced prolonged intrahepatic cholestasis improved by steroid-pulse therapy]. PMID- 8721121 TI - [A case of leiomyosarcoma of the pancreas]. PMID- 8721122 TI - [An AIDS case of probable pentamidine-induced diabetic ketoacidosis and severe acute pancreatitis]. PMID- 8721123 TI - [Stress hormone response during midazolam/fentanyl anesthesia combined with epidural anesthesia for abdominal total hysterectomy]. AB - Stress hormone response was investigated during midazolam/fentanyl/oxygen/air anesthesia combined with epidural anesthesia in six patients undergoing abdominal total hysterectomy (group MF). A control group of six patients received nitrous oxide/oxygen/sevoflurane anesthesia combined with epidural anesthesia (group C). We evaluated plasma concentrations of ACTH, cortisol, aldosterone and beta endorphin during the control period, 10 min, 60 min after incision, and in the recovery room. In group C, plasma concentrations of ACTH, cortisol, aldosterone and beta-endorphin increased significantly 60 min after incision compared with control concentrations. But in group MF, they did not increase significantly during anesthesia. In the recovery room there were significant differences compared with preinduction concentrations in both groups. It is concluded that midazolam/fentanyl anesthesia combined with epidural anesthesia is more effective than nitrous oxide/oxygen/sevoflurane anesthesia combined with epidural anesthesia to control the stress response to surgery. PMID- 8721124 TI - [The effects of sevoflurane and isoflurane on hepatic blood flow in man]. AB - The influences of sevoflurane and isoflurane on hepatic blood flow (HBF) in man were investigated employing the method of continuous infusion of indocyanine green. HBF and cardiac output (CO) were measured in patients before and after induction of anesthesia with thiamylal, fentanyl, and N2O, and again during isoflurane (n = 9) or sevoflurane (n = 8) anesthesia before the start of surgery. HBF after administration of volate anesthetics increased significantly with 1MAC isoflurane, and decreased with 1 MAC sevoflurane. Although CO decreased similarly in both groups during anesthesia, the HBF/CO ratio in isoflurane group increased significantly, and the ratio in sevoflurane group did not change significantly. We conclude that whereas isoflurane increases HBF, sevoflurane does not change it in anesthetized patients. PMID- 8721125 TI - [Association of postoperative brain dysfunction and atherosclerosis, intraoperative rSO2 and CO2 reaction in open heart surgery]. AB - We studied the relationship between postoperative brain dysfunction and the state of atherosclerosis in the patients of open heart surgery, by analyzing the intraoperative cerebral oxygen saturation (rSO2) and the CO2 reaction. The subjects were 143 patients with average age of 64 years. The patients with low postoperative Hasegawa's Dementia Score were categorized as brain dysfunction group. rSO2 was utilized to monitor cerebral blood flow, and AI and stiffness parameter beta were used for evaluation of systemic atherosclerosis. Postoperative brain dysfunction was confirmed in 12%. There were significantly high values of AI and beta as well as low rSO2, in elder age with low cardiac index and no correlation was observed between rSO2 and PaCO2 in the brain dysfunction group. The results suggested that a higher level of atherosclerosis is associated with the postoperative brain dysfunction with resultant decreased cerebral blood flow and disturbed reaction to CO2 of cerebral blood vessels. PMID- 8721126 TI - [Pharmacokinetics of propofol during liver transplantation in small pigs]. AB - A study using 14C propofol showed that the liver is the main eliminating organ for the agent. The current study was designed to clarify pharmacokinetics of propofol during liver transplantation in pigs. Five small pigs weighing 25.4 +/- 2.5 kg were anesthetized with isoflurane (0.5-1.5%) and mechanically ventilated under muscle paralysis with pancuronium. In the anhepatic phase, veno-veno bypass was placed from the inferior vena cava and portal vein to the superior vena cava. We studied pharmacokinetic parameters following an intravenous bolus injection of propofol at 2 mg.kg-1 in each phase, i.e. the pre-anhepatic, anhepatic and post anhepatic phase during liver transplantation. Pharmacokinetic analysis showed that total plasma clearance of propofol in the anhepatic phase was significantly lower than that in the post-anhepatic phase. The results suggest that propofol may be metabolized extrahepatically and can be used at reduced doses in the anhepatic phase during liver transplantation. PMID- 8721127 TI - [The relief of postoperative pain by suppositories of buprenorphine or NSAID]. AB - We evaluated the effects of suppositories of buprenorphine (BN) or NSAID (supp.) preoperatively administered for postoperative pain relief in patients who underwent elective gynecological surgeries. Fifty six patients were randomized into four groups: group B; 0.4 mg BN supp., group B+I; 0.4 mg BN supp. and 50 mg indomethacine (IND) supp., group B+D; 0.4 mg BN supp. and 50 mg diclofenac supp., group C; no supp. given as control. They were administered rectally after induction of general anesthesia. In all the supp. groups the patients had good pain relief during the first 24 hrs after the administration of supp. Group B+I seemed to have better pain relief, but, there was no statistical significance among the 3 groups. Nausea and vomiting were observed more frequently in group B and in control group C than in NSAIDs combined groups. The difference in the incidence rates was not significant. In conclusion, the simultaneous administration of BN and IND supp. was considered to be useful for postoperative pain relief without producing major side effects. PMID- 8721128 TI - [Effects of prostaglandin E1 on plasma cytokine levels during pneumonectomy]. AB - We investigated the effect of prostaglandin E1 (PGE1) on intraoperative cytokine responses and the incidence of postoperative complications. Twenty-six patients undergoing elective pneumonectomy were randomly allocated into PGE1 group (n = 12) and control group (n = 14). The PGE1 group received continuous infusion of PGE1 during surgery at a dose of 0.02-0.03 microgram.kg-1.min-1. Blood samples were obtained after induction of general anesthesia, one and two hours after incision, and immediately after the end of surgery to measure the plasma levels of tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6) and interleukin 8 (IL-8). Levels of CRP for two days after the surgery were measured and postoperative complications were recorded. Levels of TNF-alpha rose from 1.6 pg.ml-1 (mean) to 4.8 pg.ml-1 two hr after incision in the control group, while the level was suppressed in the PGE1 group (P < 0.05). No significant difference was found in IL-6 levels between the two groups. The IL-8 increased during surgery in both groups but the increase was significantly less in the PGE1 group (P < 0.05). There was no difference in CRP, and no severe postoperative complication was observed. We conclude that PGE1 administration suppresses TNF alpha and IL-8 responses during pneumonectomy, but its effects on IL-6 and the postoperative status were not significant. PMID- 8721129 TI - [Effect of sevoflurane on release of TNF-alpha and IL-1 beta from human monocytes]. AB - Although isoflurane inhibits TNF-alpha and IL-1 beta release from human monocytes stimulated by LPS in dose dependent fashion, it is unclear whether sevoflurane has the same effects. Therefore, we investigated whether sevoflurane could inhibit TNF-alpha and IL-1 beta secretions from human monocytes stimulated by LPS in dose dependent fashion in vitro. Human monocytes stimulated by LPS were cultured for 3 h in the presence of sevoflurane 1% or 5%. Another group of human monocytes were cultured in the absence of sevoflurane. TNF-alpha and IL-1 beta increased after stimulation of LPS and these increases were not inhibited by sevoflurane in a dose dependent fashion. We conclude that sevoflurane does not inhibit TNF-alpha and IL-1 beta release from monocytes stimulated by LPS. PMID- 8721130 TI - [Changing compatibility by temperature of local anesthetics with sodium bicarbonate]. AB - We investigated the amount of 7% sodium bicarbonate which could be added to local anesthetics (bupivacaine, mepivacaine and lidocaine) without any precipitation when temperature was altered (5 degrees C, 25 degrees C, 40 degrees C). Precipitations occurred at any temperatures and with any dose of bupivacaine. Interestingly, the higher the temperature, the more frequently precipitation with mepivacaine and lidocaine occurred. This was probably because the solubility of the precipitate produced by the interaction of the base of local anesthetics with bicarbonate was reduced when the temperature increased. PMID- 8721131 TI - [The influence of local anesthetics on human leukocyte functions studied by micro whole blood collection and flowcytometry]. AB - The influence of local anesthetics (pure bupivacaine and lidocaine with no preservative) on human leukocyte functions was examined. (a) The effect of bupivacaine on the phagocytosis of granulocyte was studied by bioassay. (b) The effect of lidocaine on the appearance of iC3b receptor (CR3) of granulocyte and monocyte (which is an important cell-adhesion-factor) was examined using flowcytometry. (c) The influence of lidocaine on phagocytosis of granulocyte and monocyte and on respiratory burst of granulocyte was examined using flowcytometry. (d) The influence of lidocaine on phagocytosis and that on respiratory burst were compared. These studies revealed that both phagocytosis and respiratory burst were inhibited by lidocaine, and the inhibition of respiratory burst was stronger than the inhibition of phagocytosis by local anesthetics' immunosuppressive effects. It was concluded that the balance of immunosuppressive action due to antimicrobial action and bactericidal ability of local anesthetics determined the occurrence of local bacterial infection. PMID- 8721132 TI - [Accurate placement of central venous catheters using right atrial electrocardiography]. AB - We have evaluated the effectiveness of central venous catheter placement using right atrial electrocardiography (RAECG). Consecutive patients under general anesthesia (n = 42) who required a central venous catheter underwent RAECG-guided catheter insertion procedure via right internal jugular vein. Catheter tip position was verified by post procedure portable chest radiography. Forty of 42 catheter tips were placed above the superior vena cava-right atrial junction, and none of them had its associated complications. The average insertion depth of catheters was 16.4 cm. We also attempted to predict the optimal catheter insertion depth for each patient from the previous measurements of external landmarks, but it was found to be difficult to predict reliably. In this point of view, we should use RAECG technique to make sure the proper positioning of the catheter tip. PMID- 8721133 TI - [Severe laryngeal edema immediately after extubation in a 93 year old female]. AB - A 93-year-old female, who had post-operative respiratory insufficiency, was treated with artificial ventilation for 8 days. Immediately after extubation, dyspnea, cyanosis and unconsciousness occurred. Severe laryngeal edema was found by bronchofiberscopy and reintubation seemed impossible. She underwent emergency tracheotomy and barely survived the critical state. It should be kept in mind that severe laryngeal edema might develop immediately after long term intubation in supergeriatric patients associated with hyponutrition. PMID- 8721134 TI - [Anesthetic management for colon resection in a patient with polymyositis]. AB - A patient was a 67-year-old female, 153 cm tall and weighing 47 kg. In 1988, she noticed sudden hypotonicity of her extremities, which led her to visit our hospital. Diagnosis of polymyositis (PM) was made. Treatment started with prednisolone 60 mg.day-1, followed by 2.5-5mg.day-1 as a maintenance dose. Her clinical symptoms were alleviated. However, in October, 1994, colon cancer was found and she underwent sigmoidectomy. For anesthesia, thoracic epidural block (Th 11/12) was performed. No problems occurred during and after the surgery. The following points must be considered for anesthesia of a patient with PM: (1) enhanced or delayed effect of muscle relaxant, (2) pulmonary complications- aspiration pneumonia and lung fibrosis, (3) cardiomyopathy--arrhythmia and cardiac failure, (4) steroid supplementation. In our case, because cardio pulmonary functions were almost normal, epidural anesthesia without using muscle relaxant was a successful method. PMID- 8721135 TI - [The effect of alpha-glucosidase inhibitor on burning epigastralgia in dumping syndrome: a case report]. AB - Dumping syndrome results from a rapid passage of carbohydrate in the small intestine after gastric surgery. A 50-year-old man developed the syndrome after surgery for peptic ulcer. He often complained of burning epigastralgia after meals. Various methods of treatment had only a limited effect. Acarbose, alpha glucosidase inhibitor, suppresses the breakdown of carbohydrates in the small intestine and consequently reduced osmolarity. The patient had a dramatic improvement in the dumping syndrome including epigastralgia, diarrhea and perspiration with an administration of acarbose 50 mg. There was no intractable side effect. Preprandial administration of acarbose is a reasonable treatment in the case of dumping syndrome. PMID- 8721136 TI - [Three cases of hyperkalemia after reperfusion associated with revascularization for massive acute arterial occlusions]. AB - We encountered three cases of hyperkalemia after the reperfusion of ischemic limbs associated with revascularization surgery for acute arterial occlusions. Two patients died because of uncontrollable hyperkalemia in spite of intravenous insulin and forced diuresis. Careful attention should be paid to hyperkalemia and metabolic acidosis for several hours after the reperfusion in cases of ischemia and/or extensive ischemia of long duration. The aggressive prophylaxis and treatment for MNMS are vital to the anesthetic management of revascularization for acute arterial occlusions. PMID- 8721137 TI - [Anesthetic and postoperative care of a patient with Crigler-Najjar syndrome type II]. AB - Crigler-Najjar syndrome is a rare inherited deficiency of bilirubin uridine diphosphate glucuronyl transferase, characterised by lifelong unconjugated hyperbilirubinemia. We report anesthetic and postoperative management of a patient with this syndrome. A 64-yr-old man with Crigler-Najjar syndrome type II underwent cholecystectomy. Preoperative laboratory values included total bilirubin 8.4 mg.dl-1 and direct-bilirubin 1.8 mg.dl-1. He received epidural anesthesia combined with general anesthesia with nitorous oxide and isoflurane. He had no remarkable perioperative complications. PMID- 8721138 TI - [Use of a laryngeal mask airway for anesthesia in a patient with bronchomalacia]. AB - A 56-year-old woman with bronchomalacia underwent three consecutive operations for bronchoscopy, cautery YAG-laser, and the insertion of a stent under general anesthesia using a laryngeal mask airway (LMA). For the first operation, anesthesia was induced with ketamine 20 mg, diazepam 5 mg and pentazocine 20 mg. The patient was ventilated with N2O-O2-sevoflurane with a face mask. Then vecuronium 6 mg was administered intravenously and LMA was inserted blindly. Anesthesia was maintained with N2O-O2-sevoflurane during bronchoscopy. Immediately after the operation, the patient coughed and experienced dyspnea. The symptoms were alleviated using theophyllin, hydrocortisone and droperidol. For the second and third operations, anesthesia was induced with droperidol and fentanyl with N2O-O2. After administration of vecuronium, LMA was inserted. The patient was stable during the second and third operations compared with the first operation. It was concluded that LMA may be useful for anesthetic management of a patient with bronchomalacia. PMID- 8721139 TI - [Cardiac output in patients in their 90's]. AB - Cardiac output was measured by the thermodilution method in nine patients aged 90 to 97 years. The patients were capable of walking in their daily lives before the injury or admission to the hospital, after which they were scheduled for operations. The causes for operations were localized and did not markedly influence general conditions of the patients. After heart rate, arterial blood pressure and mental condition of the patients had been stabilized, their cardiac outputs were determined before the induction of anesthesia. The results are as follows: Cardiac output: 3.58 +/- 0.90 l.min-1, cardiac index: 2.72 +/- 0.57 l.min-1.m-2. Previously published reports in Japan and the results of this study about the relationship between cardiac index and age suggest that as a whole there is a tendency for the cardiac index to decrease with aging. PMID- 8721140 TI - [Intravenous infusion of prostagrandin E1 during and after gastrectomy on postoperative hepato-renal functions in senile patients]. AB - Effect of intravenous prostagrandin E1 (PGE1) during and after surgery on postoperative hepatorenal functions in senile patients was evaluated in 36 elective surgical patients ranged in age from 60 to 85 years. The patients with carcinoma of the stomach underwent total or subtotal gastrectomy under isoflurane anesthesia. These patients were devided into two groups. Eighteen patients received intravenous PGE1 at a rate of 0.03-0.13 micrograms.kg-1.min-1 during surgery and 0.03 microgram.kg.min-1 after surgery until 9:00 am on the first operative day. The remaining 18 patients did not receive PGE1 and served as the control group. Serum GOT and GPT levels in both groups increased significantly at emergence from anesthesia compared with the preanesthetic levels and then declined to the preanesthetic levels on the 3rd postoperative day. Thereafter they increased significantly again on the 7th postoperative day. Serum GOT levels in the PGE1 administered group were significantly lower than those in the control group at the emergence from anesthesia. Serum GPT levels in the PGE1 group tended to be lower than those in the control group on awakening from anesthesia and on the first postoperative day. Serum gamma-GTP levels were stable postoperatively but they increased significantly on the 7th postoperative day in both groups. Serum bilirubin levels were within normal limits in both groups. Postoperative serum levels of urea nitrogen and creatinine were at the preanesthetic levels in both groups. Our findings suggest that continuous intravenous administration of PGE1 during and after surgery is beneficial in attenuating hepatic injury in senile patients for gastrectomy. However, protective effect of PGE1 on postoperative renal function was found to be vague in this study. PMID- 8721141 TI - [Comparison of the effects of halothane, isoflurane and sevoflurane in the treatment of severe asthmatic attack: a case report]. AB - A 19-year-old woman suffered from a severe asthmatic attack. She did not respond to any of the conventional therapies and eventually lapsed into a state of unconsciousness. We administered isoflurane 1-2% via an endotracheal tube, but, she still required a ventilator. This study was carried out in order to compare the effects of halothane, isoflurane and sevoflurane in the treatment of a severe asthmatic patient. The inhalation of isoflurane, halothane and sevoflurane decreased the airway resistance and systemic vascular resistance in direct relationship to the dosage administered. Because the patient's metabolic rate is low and the occurrence of arrhythmias is rare with isoflurane, we concluded that isoflurane is the best alternative for the patient with asthma. PMID- 8721142 TI - [Selective epidural nerve root block using a superfine fiberscope]. AB - A 65 yr-old male with severe scar pain on his right lower abdomen underwent selective nerve block through epidural space using a superfine fiberscope. A12 gauge Tuohy needle (internal diameter 2.2 mm) was inserted at the intervertebral space of Th12-L1. The fiberscope (external diameter 1.1 mm) was introduced through the needle and the epidural space was observed. When the dura mater of Th12 nerve root level was seen, the patient had pain on the scar area. Then 2% lidocaine 1.5 ml and 60% megumine sodium amidotrizoate 1.5 ml were slowly injected. Soon after injection of anesthetic, analgesia area (Th11-L1) was obtained, and Th12 nerve root was identified by X ray. Epidural blockade using a superfine fiberscope seems to be one of the useful and reliable methods for the selective nerve blockade. PMID- 8721143 TI - [Postoperative analgesic requirements after various operations]. AB - The difference in postoperative analgesic requirements for operations involving 11 sites was assessed in 1100 patients. Pain relief was mainly attained by epidural administration of bupivacaine 0.25% on demand. Postoperative analgesic was required most frequently after thoracotomy, followed by gastrectomy with left diagonal approach, gastrectomy with median approach, operation for colon cancer, cholecystectomy and hysterectomy. After hip operation, periproctal operation, radical mastectomy, appendectomy and herniorrhephies, total analgesic requirement was similar and the minimum amount required. In conclusion, there was a high degree of correlation between the severity of the pain and the site of the operation. PMID- 8721144 TI - [Survey on anesthesia related accident in 1993]. PMID- 8721145 TI - Analysis of the structural heterogeneity of laminarin by electrospray-ionisation mass spectrometry. AB - Electrospray-ionisation-mass spectrometry (ESIMS) was used in conjunction with chemical derivatisation and degradation procedures to analyse the size heterogeneity and branching structure of laminarin from the brown alga, Laminaria digitata. Laminarin is a beta-(1-->3)-linked D-glucan with occasional beta-(1- >6)-linked branches. Electrospray-ionisation-mass spectrometry of permethylated laminarin distinguished two homologous series of molecules, a minor G-series containing 22-28 glucosyl residues, and a more abundant M-series containing 20-30 glucosyl residues linked to a mannitol residue. The relative abundance of all these molecular species could be determined simultaneously from a single mass spectrum, with a mean mass error of 0.6 atomic mass units and a mean mass accuracy of 0.011%. Both series had a mean degree of polymerisation of 25 glucosyl residues, and an approximately 3:1 molar ratio of M-series to G-series molecules was maintained across the range of molecular sizes. Treatment of laminarin with periodate, followed by reduction with borohydride, degraded terminal glucosyl residues on both the main chain and the branches, and allowed the detection of isomers differing solely in their degree of branching. M-series molecules were thus shown to contain 0, 1, 2, 3 or 4 branches, with an average of 1.3 branches per molecule; branched G-series molecules were also detected. Subsequent treatment with acid (Smith degradation) showed that 75% of the branches were single glucosyl residues. This study thus shows how the speed, resolution and mass accuracy of electrospray-ionisation-mass spectrometry can be used in the detailed structural analysis of a polydisperse polysaccharide. PMID- 8721146 TI - Mass spectrometric studies of the thermal decomposition of carbohydrates using 13C-labeled cellulose and glucose. AB - The mechanism of the thermal decomposition of carbohydrates is very important to the development of fuels, fibers, and paper products. To help gain more insight into the pyrolysis chemistry of cellulose, we have carried out experimental studies using Acetobacter xylinum cellulose grown on D-(1-13C)-glucose medium with incorporation levels of 1-13C of 14%, as determined by 13C NMR analysis. Samples of the labeled cellulose, as well as D-(1-13C)- and D-(2-13C)-glucose, were pyrolyzed under fast-heating conditions and the products analyzed by molecular beam mass spectrometry (MBMS). From the labeled cellulose samples, statistically significant levels of enrichment were observed for the pyrolysis products that occur at m/z 110, 114, 126, 144, and 191, but not at m/z 98, 60, or 31. The lack of enrichment in the latter fragments indicates that they do not incorporate C-1. Samples were treated with 0.1% aqueous KOH to favor the formation of glycolaldehyde, but even in this case this major product was not enriched. These results suggest that the m/z 60 ion is an EI fragment ion of levoglucosan, formed by loss of neutral species containing C-1. However, the m/z 191 ion was found to contain two C-1 carbons. Collision-induced dissociation results for this ion suggest that it consists of a formate group. The structure of this ion is proposed to be the protonated formate of levoglucosan at the C-4 position, derived from either a reverse aldol reaction or a (+/-)-Diels-Alder reaction. The (1-13C)-glucose pyrolysis product distribution is similar to that for (1-13C)-labeled cellulose. The (2-13C)-glucose pyrolysis product distribution shows significant contribution from C-2 at the M + 1 peaks of m/z 32, 43, 60, 73, 85, and 97, indicating major incorporations of C-2 in glycolaldehyde in contrast to the lack of incorporation of the C-1 position in this major product. PMID- 8721147 TI - Total synthesis of VIM-2 ganglioside isolated from human chronic myelogenous leukemia cells. AB - A total synthesis of the tumor-associated glycolipid antigen, VIM-2, is described [2]. Phenyl 2,3,4-tri-O-benzoyl-6-O-benzyl-beta-D-galactopyranosyl-(1-->4)-6-O benzy l-2- deoxy-2-phthalimido-1-thio-beta-D-glucopyranoside (7), a key intermediate prepared by condensation of phenyl 6-O-benzyl-2-deoxy-2-phthalimido 1-thio-beta-D-glucopyranoside (6) and 2,3,4-tri-O-benzoyl-6-O-benzyl-alpha-D galactopyranosyl bromide (5), was glycosylated with methyl 2,3,4-tri-O-benzyl-1 thio-beta-L-fucopyranoside (8) to give the trisaccharide donor 9, which, on coupling with 2-(trimethylsilyl)ethyl 2,4,6-tri-O-benzyl-beta-D-galactopyranosyl (1-->4)-2,3,6-tri-O-benzyl-be ta-D- glucopyranoside (10), afforded the pentasaccharide 11. The regioselective glycosylation of 12 (derived by O debenzoylation of 11) with 7 gave the heptasaccharide 13, which was converted by treatment with hydrazine monohydrate and subsequent N-acetylation into the hexasaccharide acceptor 14. The stereo- and regio-selective glycosylation of 14 with methyl (phenyl 5-acetamido-4,7,8,9-O-benzoyl-3,5-dideoxy-2-thio-D-glycero beta-D-galact o-2- nonulopyranosid)onate (16) gave the desired octasaccharide 18. Hydrogenolytic removal of the benzyl groups in 18 and successive O-acetylation, removal of the 2-(trimethylsilyl)ethyl group, and treatment with trichloroacetonitrile gave the alpha-trichloro-acetimidate 21, which was then coupled with (2S,3R,4E)-2-azido-3-O-(tert-butyldiphenylsilyl)-4-octade cene-1,3 diol (22) to give 23. Compound 23 was transformed, via selective reduction of the azido group, N-introduction of octadecanoic acid, O-desilylation, O-deacylation, and saponification of the methyl ester group, into the title VIM-2 ganglioside 26. PMID- 8721148 TI - L-iduronic acid derivatives as glycosyl donors. AB - O-[Methyl (2-O-acetyl-3-O-benzyl-4-O-levulinyl-alpha, and beta-L idopyranosid)uronate] trichloroacetimidate and the corresponding n-pentenyl glycosides are efficient L-iduronic acid glycosyl donors. Both have been used for the high-yielding synthesis of basic disaccharide blocks which are useful for the subsequent synthesis of complex oligosaccharides related to heparin/heparan sulfate, and dermatan sulfate. In contrast, the corresponding thioethyl glycosides, thiophenyl glycosides, and fluoride, did not yield the expected disaccharides. PMID- 8721149 TI - Structure of a galactan from cell walls of Bifidobacterium catenulatum YIT4016. AB - A structural study was carried out on a galactose-rich polysaccharide fraction isolated from cell walls of Bifidobacterium catenulatum YIT4016 after N acetylmuramidase digestion. The polysaccharide contained galactose and glucosamine in a molar ratio of 16.9:1.0. Data obtained by 13C NMR spectroscopy showed that the backbone chain of this polysaccharide is composed of galactofuranose residues, while the branches consist of galactopyranosyl residues. Furthermore, the data obtained from NaIO4 oxidation, partial methanolysis and methylation analysis indicated that this polysaccharide consists of a trisaccharide repeating unit having the following structure: [sequence: see text] PMID- 8721150 TI - Somatosensory and movement-related properties of red nucleus: a single unit study in the turtle. AB - Extracellular recordings were performed from turtle red nucleus neurons to examine their responsiveness to peripheral somatic stimulation and to study differences between rubral sensory and movement-related responses. In pentobarbital sodium-anesthetized or decerebrate turtles, red nucleus neurons could be divided into two categories based on their response characteristics. The first group, which included 87% of neurons studied, had low spontaneous rates of activity and responded with excitation to electrical stimulation of the spinal cord or the cerebellum, or during active movement of the contralateral limbs. Neurons in this category were likely to be rubrospinal cells. The remaining 13% of cells studied had higher rates of spontaneous discharge and were inhibited by electrical stimulation or during active movement. These cells might be rubral GABAergic interneurons. Single red nucleus neurons responded with excitation and/or inhibition to somatosensory stimulation. Unlike the motor fields, which were restricted to a single contralateral limb, red nucleus sensory receptive fields were wide and often bilaterally distributed. Rubral responsiveness to sensory stimulation was found to be significantly diminished during active limb movements, thereby suggesting that sensory inputs to the red nucleus are not used for the on-line modification of motor commands. Inactivation of the cerebellar cortex enhanced the sensory responsiveness of rubral neurons and expanded the size of red nucleus receptive fields. These results suggest that the red nucleus receives substantial sensory input, and that the cerebellar cortex can modify the flow of sensory information to the red nucleus. PMID- 8721151 TI - Previous heat shock treatment attenuates bicuculline-induced convulsions in rats. AB - Exposure to elevated temperature provokes a sequence of events (heat shock response) in all living organisms. Through this response, heat shock proteins (HSPs) are induced and protect the cells against subsequent injury. We investigated the effect of heat treatment on bicuculline-induced convulsions, and analyzed a possible role of HSPs. Screw electrodes were implanted in the brain of mature male Wistar rats for electroencephalogram (EEG) recording. Experimental rats were subjected to whole-body hyperthermia at 41-42 degrees C for 15 min. Fifteen hours later, bicuculline was injected intraperitoneally to induce convulsions in both experimental and control groups. The heated rats showed a significant attenuation of the convulsive response, in terms of both spike discharges in EEG and clinical seizures. Further-more, induction of HSP72 was detected in the brain of heat-treated rats by immunoblotting, appearing at 4 h and reaching a maximal level 16-24 h after the heat shock. We conclude that the previous heat treatment stabilized neuronal excitability, most probably through the induction of HSP72. PMID- 8721152 TI - Contribution of the forebrain archistriatal gaze fields to auditory orienting behavior in the barn owl. AB - A region in the barn owl forebrain, referred to as the archistriatal gaze fields (AGF), is shown to be involved in auditory orienting behavior. In a previous study, electrical microstimulation of the AGF was shown to produce saccadic movements of the eyes and head, and anatomical data revealed that neurons in the AGF region of the archistriatum project directly to brainstem tegmental nuclei that mediate gaze changes. In this study, we investigated the effects of AGF inactivation on the auditory orienting responses of trained barn owls. The AGF and/or the optic tectum (OT) were inactivated pharmacologically using the GABAA agonist muscimol. Inactivation of the AGF alone had no effect on the probability or accuracy of orienting responses to contralateral acoustic stimuli. Inactivation of the OT alone decreased the probability of responses to contralateral stimuli, but the animals were still capable of orienting accurately toward stimuli on about 60% of the trials. Inactivation of both the AGF and the OT drastically decreased the probability of responses to 16-21% and, on the few trials that the animals did respond, there was no relationship between the final direction of gaze and the location of the stimulus. Thus, with the AGF and OT both inactivated, the animals were no longer capable of orienting accurately toward acoustic stimuli located on the contralateral side. These data confirm that the AGF is involved in gaze control and that the AGF and the OT have parallel access to gaze control circuitry in the brainstem tegmentum. In these respects, the AGF in barn owls is functionally equivalent to the frontal eye fields in primates. PMID- 8721153 TI - Evidence for glutamate as a neurotransmitter in spinothalamic tract terminals in the posterior region of owl monkeys. AB - Previous studies have suggested that glutamate is a neurotransmitter in ascending somatosensory pathways to the thalamus. The present study examined with quantitative immunohistochemical methods the presence of glutamate in spinothalamic tract terminals of owl monkeys (Aotus trivirgatus). Such terminals in the posterior region, in which a nucleus was recently identified as a specific pain and temperature relay in macaques and humans, were labeled by anterograde transport of wheat germ agglutinin conjugated to horseradish peroxidase, injected into the spinal dorsal horn. Glutamate-like immunoreactivity was demonstrated with a postembedding immunogold procedure using a well-characterized glutamate antiserum. Quantitative analysis of the immunogold labeling demonstrated that the spinothalamic tract terminals contained more than twice the tissue average of glutamate-like immunoreactivity. Enrichment of glutamate-like immunoreactivity was also found in terminals of presumed cortical origin. Presynaptic dendrites, cell bodies and non-vesicle-containing dendrites displayed low levels of glutamate-like immunoreactivity. A strong positive correlation (r = 0.69; P < 0.0001) was found between the density of synaptic vesicles and the density of gold particles in spinothalamic tract terminals, in contrast to a weak negative relationship (r = -0.28; P = 0.089) present in GABAergic presynaptic dendrites. These data provide strong evidence that the gold labeling in the spinothalamic tract terminals represents transmitter labeling, implying that glutamate is a neurotransmitter for ascending nociceptive and thermoreceptive information in primates. PMID- 8721155 TI - Sex differences in the developmental changes of GABAergic neurons in zebra finch song control nuclei. AB - Small neurons in a sexually dimorphic nucleus of the zebra finch, the robust nucleus of the archistriatum (RA), were immunoreactive for gamma-aminobutyric acid (GABA). This dimorphism arose from the sex differences in the development of the RA neurons. We examined whether neurons with GABA-like immunoreactivity (GABA LI) undergo the same developmental changes as the cresyl-violet-stained neurons in the RA. Somata with GABA-LI from male finches became larger during development, while somata with GABA-LI from female finches became smaller, at a rate similar to that in the total population of RA neurons. However, there were marked sex differences in the development of the number of RA neurons with GABA LI. In the female, neurons with GABA-LI were lost at a rate which was similar to the rate of loss of the total population of RA neurons. On the other hand, in the male, the number of neurons with GABA-LI increased transiently during the sensitive period of song learning, while the total number of RA neurons did not change. This transient increase may be related to the fact that only male birds learn their song during a sensitive period. PMID- 8721154 TI - Differential relation of discharge in primary motor cortex and premotor cortex to movements versus actively maintained postures during a reaching task. AB - The activity of cells in primary motor cortex (MI) and dorsal premotor cortex (PMd) were compared during reaching movements in a reaction-time (RT) task, without prior instructions, which required precise control of limb posture before and after movement. MI neurons typically showed strong, directionally tuned activity prior to and during movement as well as large gradations of tonic activity while holding the limb over different targets. The directionality of their movement- and posture-related activity was generally similar. Proximal-arm muscles behaved similarly. This is consistent with a role for MI in the moment-to moment control of motor output, including both movement and actively maintained postures, and suggests a common functional relation for MI cells to both aspects of motor behavior. In contrast, PMd cells were generally more phasic, frequently emitting only strong bursts of activity confined mainly to the behavioral reaction time before movement onset. PMd tonic activity during different postures was generally weaker than in MI, and showed a much more variable relation with their movement-related directional tuning. These results imply that the major contribution of PMd to this RT task occurred prior to the onset of movement itself, consistent with a role for PMd in the selection and planning of visually guided movements. Furthermore, the nature of the relative contribution of PMd to movement versus actively maintained postures appears to be fundamentally different from that in MI. Finally, there was a continuous gradient of changes in responses across the rostrocaudal extent of the precentral gyrus, with no abrupt transition in response properties between PMd and MI. PMID- 8721156 TI - Time course of striatal changes induced by 6-hydroxydopamine lesion of the nigrostriatal pathway, as studied by combined evaluation of rotational behaviour and striatal Fos expression. AB - Changes taking place after unilateral 6-hydroxydopamine lesion of the dopaminergic nigrostriatal system have been studied by performing spontaneous, amphetamine-induced and apomorphine-induced rotational behaviour testing and tyrosine hydroxylase (TH) and Fos protein immunohistochemistry in the same rats. Apomorphine at a low dosage (0.25 mg/kg) induced contraversive rotation and supersensitive striatal Fos expression that were detected 24-48 h post-lesion and gradually increased in magnitude. Twenty-four hours after lesion, both high (5 mg/kg) and low doses (0.5 mg/kg) of D-amphetamine induced contraversive rotation and intense striatal Fos activation on the denervated side; however, only the higher dose induced Fos on the normal side. Two, 3 and 4 days after lesion, 0.5 mg/kg amphetamine induced contraversive rotation, but 5 mg/kg induced transitory contraversive rotation which switched to ipsiversive. In the normal striatum, only high doses of amphetamine induced Fos, but Fos induction in the denervated striatum was similar with both doses: areas showing severely decreased TH immunoreactivity still showed considerable Fos immunoreactivity, and some areas still showing TH immunoreactivity had higher Fos density than in the normal side. Seven and 14 days after lesion the loss of TH immunoreactivity and apomorphine induced supersensitive Fos expression were more evenly distributed, and amphetamine induced only ipsiversive rotation and a low density of Fos-positive nuclei in the denervated striatum. These results indicate that the severe and progressive loss of dopaminergic terminals is counteracted by an early and rapidly progressing dopamine supersensitivity, together with a higher susceptibility to drug-induced dopamine release. This explains the apparently paradoxical contraversive rotation induced by amphetamine during the first week post lesion. However, experiments involving successive drug injections indicated that only the first amphetamine injection releases dopamine from the lesioned terminals. PMID- 8721157 TI - Spatial coordination by descending vestibular signals. 2. Response properties of medial and lateral vestibulospinal tract neurons in alert and decerebrate cats. AB - Spatial response properties of medial (MVST) and lateral (LVST) vestibulospinal tract neurons were studied in alert and decerebrate cats during sinusoidal angular rotations of the whole body in the horizontal and many vertical planes. Of 220 vestibulospinal neurons with activity modulated during 0.5-Hz sinusoidal rotations, 200 neurons exhibited response gains that varied as a cosine function of stimulus orientation and phases that were near head velocity for rotation planes far from the minimum response plane. A maximum activation direction vector (MAD), which represents the axis and direction of rotation that maximally excites the neuron, was calculated for these neurons. Spatial properties of secondary MVST neurons in alert and decerebrate animals were similar. The responses of 88 of 134 neurons (66%) could be accounted for by input from one semicircular canal pair. Of these, 84 had responses consistent with excitation from the ipsilateral canal of the pair (13 horizontal, 27 anterior, 44 posterior) and 4 with excitation from the contralateral horizontal canal. The responses of the remaining 46 (34%) neurons suggested convergent inputs. The activity of 38 of these was significantly modulated by both horizontal and vertical rotations. Twelve neurons (9%) had responses that were consistent with input from both vertical canal pairs, including 9 cells with MADs near the roll axis. Thirty-two secondary MVST neurons (24%) had type II yaw and/or roll responses. The spatial response properties of 18 secondary LVST neurons, all studied in decerebrate animals, were different from those of secondary MVST neurons. Sixteen neurons (89%) had type II yaw and/or roll responses, and 12 (67%) appeared to receive convergent canal pair input. Convergent input was more common on higher-order vestibulospinal neurons than on secondary neurons. These results suggest that MVST and LVST neurons and previously reported vestibulo-ocular neurons transmit functionally different signals. LVST neurons, particularly those with MADs close to the roll axis, may be involved in the vestibular-limb reflex. The combination of vertical and ipsilateral horizontal canal input on many secondary MVST neurons suggests a contribution to the vestibulocollic reflex. However, in contrast to most neck muscles, very few neurons had maximum vertical responses near pitch. PMID- 8721158 TI - Calretinin-immunoreactivity in organotypic cultures of the rat cerebral cortex: effects of serum deprivation. AB - Calretinin, a calcium-binding protein, is expressed in a specific set of interneurons in the adult rat cortex. Although its role in development is not known with any degree of certainty, evidence in support of a neuroprotective function has been forthcoming. To test this hypothesis, we submitted organotypic cultures (interphase technique) of 4- to 6-day-old rat brain slices to nutritive stress by serum deprivation for 1-3 weeks. Cultures were immunolabelled either with an antiserum against calretinin or with an antibody against MAP2 (the latter being used to assess neuronal cell number). In control (serum-enriched) cultures, the pattern of development of calretinin immunoreactivity mimicked that evinced in vivo with respect to layer- and cell-type specificity, but the maturation process was retarded by about 1 week. In the experimental group, cultures were incubated for 1 week in the presence of serum and then transferred to serum-free medium for an additional 2 weeks. Tissue was characterized by necrotic foci, a marked decrease in neuronal cell number and a further retardation in the course of development of the calretinin immunoreactivity pattern. The proportion of calretinin-immunoreactive cells to total number of viable neurons was 16% in serum-free cultures as against 9% in serum-enriched ones, suggesting that cells expressing the calcium-binding protein exhibit a greater tenacity for survival under conditions of nutritive stress, and thereby supporting the contention that calretinin acts in a neuroprotective capacity. PMID- 8721159 TI - Changes in hippocampal cell discharge patterns and theta rhythm spectral properties as a function of walking velocity in the guinea pig. AB - Dorsal hippocampal theta rhythm (theta) and extracellular unit activity from CA1 pyramidal layer were recorded in awake guinea pigs, both during standing and during walking on a conveyor belt at increasing speeds. Amplitude, frequency and rhythmicity of theta increased linearly with the movement speed. In this preparation we found the same three types of unit discharge patterns that have been described in anesthetized rats in the presence of spontaneous or induced hippocampal theta: type 1, rhythmic at theta frequency and phase-locked with theta; type 2, discharging non-rhythmically but phase-locked with theta; and type 3, discharging at random. Furthermore, all units modified their firing pattern when the animals walked, either by increasing their rhythmicity and/or phase locking with theta or by increasing their firing frequency. During walking, some type 3 units changed into type 2 or type 1, type 2 units changed to type 1, and type I increased their rhythmicity. Consequently, the unit discharge rhythmicity and phase-locking with theta increased with the speed of movement. The mean rate of neuronal discharges increased linearly as a function of walking speed. In this paper we show that the progressive spectral theta changes determined by the intensity of movement are concomitant with the increase in rhythmicity of hippocampal cells. Moreover, the firing rate of these cells, and the amplitude, frequency and rhythmicity of theta, increased linearly as a function of walking speed, suggesting that neuronal excitation may be basically responsible for these changes in theta properties. PMID- 8721160 TI - Early enhancement but no late changes of motor responses induced by intracortical microstimulation in the ketamine-anesthetized rat. AB - The objectives of this study were to determine whether changes in electromyographic (EMG) responses observed during prolonged intracortical microstimulation (ICMS) were due to local plasticity of the motor system or to global changes in the preparation. Local effects would be expressed as changes only along the activated motor pathway, whereas global effects would be expressed as changes also appearing at distant cortical efferent microzones. The results of ICMS in the ketamine-anesthetized rat showed that the size of consecutive EMG responses increased gradually to a relatively stable magnitude over a period of four to six trains of stimuli. This early enhancement of EMG responses was maintained while continuously providing trains of stimuli at 1 Hz. However, it disappeared after a 5-min period of muscle inactivity. This response enhancement in the presence of ketamine (an NMDA, N-methyl-D-aspartate, receptor blocker) suggests that a neuronal mechanism involving non-NMDA-mediated homosynaptic short term potentiation (STP) was responsible for the early enhancement of EMG responses. To compare ICMS effects at several time intervals it was necessary to average several evoked EMG responses because there was normal biological variability between single EMG responses. To determine the optimal number of EMG responses that would provide a reliable average EMG response, averages of 5, 10, 15, 20, and 25 EMG responses evoked from a single cortical site were collected at 5-min intervals. The results revealed that averages of 10 responses would provide reliable average EMG responses for all subsequent analyses. There were wide fluctuations in the average EMG responses when periodic injections of ketamine were used to maintain a low reflexive state in the animal. Switching to continuous infusion of ketamine abolished these fluctuations but there remained a small drift in the magnitudes of consecutive EMG responses. To test whether this drift reflected local plastic changes in the motor system induced by stimulation or some global changes, EMG responses evoked from another ICMS site were used as control. The rationale was that global effects would affect all motor output sites equally. The sizes of control EMG responses followed a similar time course to those evoked from the test site. Furthermore, standardizing the test EMG responses with respect to the control responses eliminated the drift in response magnitudes. Thus the drift was due to slow global changes in neuronal excitability possibly produced by the anesthesia. In conclusion, late changes occurring after hours of ICMS were not due to plasticity of the motor system but rather to global changes in the preparation, possibly resulting from the inability to set an ideal anesthetic infusion rate that could maintain a constant level of neuronal excitability over long periods of time. However, there was early enhancement of the EMG responses evoked by ICMS due to neuronal plasticity possibly mediated by a non-NMDA mechanism of homosynaptic STP such as post tetanic potentiation (PTP). This early enhancement would favor recruitment of the previously activated motor pathway and lead to greater consistency in movement execution. PMID- 8721161 TI - Availability of visual and proprioceptive afferent messages and postural control in elderly adults. AB - The ability of young and elderly adults to keep a stable upright posture while facing changes in the availability of visual and/or propriomuscular information was investigated. The two sensory sources of information were alternatively available and withdrawn, jointly and separately, during 10-s alternating sequences. Vision was modified by means of liquid-crystal goggles, and proprioception was altered by means of tendon vibration of both antagonistic ankle muscles. Elderly adults were less stable than young adults when vision was withdrawn. Both groups were greatly affected when propriomuscular inputs were altered by vibration. Under constant visual conditions and following a propriomuscular perturbation (i.e., vibration), elderly adults were unable to take advantage of the reinsertion of propriomuscular inputs. They showed a transient, decreased stability and were unable to fully recover during a 10-s period, whereas young adults were able to rapidly integrate the information to stabilize their posture. When both propriomuscular and visual inputs were withdrawn and concurrently reinserted, the elderly adults did not show a transitory increase in the velocity of the center of foot pressure. The present results extend findings on the inability of elderly adults to reconfigure rapidly the postural set following reinsertion of sensory inputs. The results also suggest that elderly adults have difficulties in taking advantage of sensory redundancy in postural control. PMID- 8721162 TI - Mechanisms for distance reproduction in perceptual and motor tasks. AB - Two experiments were carried out: the control experiment and the doubling distance experiment. In the control experiment subjects were presented with two visual stimuli whose distance was randomly varied. Subjects were required to reproduce the interstimulus remembered distance in two conditions. In one condition (reproduction by pointing) they pointed to a virtual position in space. In the other condition (visual reproduction) they matched the distance by using two other visual stimuli. In the doubling-distance experiment, distances between the two randomly presented stimuli were half of the distances used in the control experiment. Subjects were required to reproduce the double of the presented distance. As in the control experiment, reproduction was executed in two conditions: reproduction by pointing and visual reproduction. In both experiments variable and constant errors were measured. Pointing kinematics were also analysed. The results of the control experiment showed that subjects underestimated distance in reproduction by pointing, whereas they overestimated distance in visual reproduction. Variable errors increased with increasing distance, whereas they were not influenced by the type of reproduction. In the doubling-distance experiment, subjects generally overestimated distance by the same amount in both conditions. However, overestimation decreased with distance during reproduction by pointing. Pointing kinematics varied between the two experiments. The results of the control experiment confirm the hypothesis that perceptual judgement and visuo-motor transformation are two separate processes during which the same object attributes are independently analysed. However, the results of the doubling-distance experiment suggest that perceptual judgement and visuo-motor transformation use the same mechanisms when object attributes are deduced by mental elaboration. PMID- 8721163 TI - The contribution of transcortical pathways to long-latency stretch and tactile reflexes in human hand muscles. AB - Long-latency electromyographic (EMG) responses can be evoked in the first dorsal interosseous muscle (FDI) by unexpected slips of an object (skin stretch) held between the index and thumb, or by forcible adduction of the metacarpophalangeal joint (muscle stretch). The former type of response is due to stimulation of tactile afferents in the skin of the digits, whereas the latter also activates muscle receptors. Previous studies have provided good evidence that long-latency reflex responses to stretch of distal muscles involve activity in a transcortical reflex pathway. The present experiments examined whether cutaneous reflexes also utilise a transcortical route. Transcranial magnetic or electrical stimuli were given over the motor cortex to evoke EMG activity during the period of the long latency reflex response. When evoked by muscle stretch the responses to magnetic stimulation were facilitated more than those to electric stimulation. In contrast, facilitation was equal during the long-latency reflex elicited by cutaneous stimulation. Because of the different ways in which electrical and magnetic stimuli are believed to activate the motor cortex, we interpret these results to mean that the long-latency response to skin stretch is not mediated by a transcortical mechanism in the majority of subjects, whereas that following muscle stretch is. However, these are average data. In a few individual subjects, the opposite results were obtained. We suggest that there may be differences between subjects in the transcortical contribution to long-latency reflex responses. The implication is that, under normal circumstances, several pathways may contribute to these responses. If so, the relative roles of the pathways may change during different tasks, and in pathological states lesions in one system may well be accompanied by compensatory changes in other systems. PMID- 8721165 TI - Control of grip force during restraint of an object held between finger and thumb: responses of muscle and joint afferents from the digits. AB - Pulling or pushing forces applied to an object gripped between finger and thumb excite tactile afferents in the digits in a manner awarding these afferents probable roles in triggering the reactive increases in grip force and in scaling the changes in grip force to the changes in applied load-force. In the present study we assessed the possible contributions from slowly adapting afferents supplying muscles involved in the generation of grip forces and from digital joint afferents. Impulses were recorded from single afferents via tungsten microelectrodes inserted percutaneously into the median or ulnar nerves of awake human subjects. The subject held a manipulandum with a precision grip between the receptor-related digit (index finger, middle finger, ring finger or thumb) and an opposing digit (thumb or index finger). Ramp-and-hold load forces of various amplitudes (0.5-2.0 N) and ramp rates (2-32 N/s) were delivered tangential to the parallel grip surfaces in both the distal (pulling) and the proximal (pushing) directions. Afferents from the long flexors of the digits (n = 19), regardless of their muscle-spindle or tendon-organ origin, did not respond to the load forces before the onset of the automatic grip response, even with the fastest ramp rates. Their peak discharge closely followed the peak rate of increase in grip force. During the hold phase of the load stimulus, the afferents sustained a tonic discharge. The discharge rates were significantly lower with proximally directed loads despite the mean grip-force being similar in the two directions. This disparity could be explained by the differing contributions of these muscles to the finger-tip forces necessary to restrain the manipulandum in the two directions. Most afferents from the short flexors of the digits (n = 17), including the lumbricals, dorsal interossei, opponens pollicis, and flexor pollicis brevis, did not respond at all, even with the fastest ramps. Furthermore, the ensemble pattern from the joint afferents (n = 6) revealed no significant encoding of changes in finger-tip forces before the onset of the increase in grip force. We conclude that mechanoreceptors in the flexors of the digits and in the interphalangeal joints cannot be awarded a significant role in triggering the automatic changes in grip force. Rather, their responses appeared to reflect the reactive forces generated by the muscles to restrain the object. Hence, it appears that tactile afferents of the skin in contact with the object are the only species of receptor in the hand capable of triggering and initially scaling an appropriate change in grip force in response to an imposed change in load force, but that muscle and joint afferents may provide information related to the reactive forces produced by the subject. PMID- 8721166 TI - Evidence for a facilitatory role of callosal afferents to the cat motor cortex in the initiation of conditioned bilateral movements. AB - The effects of selective transection of the rostralmost portion of the corpus callosum, which contains fibres interconnecting the motor cortices of the two hemispheres, on frequency of occurrence and latency of conditioned responses (CRs) in both eyes were examined in seven cats trained to blink in response to a 500-ms tone. A 100-ms air-puff delivered to one eye only (ipsilateral eye) 400 ms after tone onset was used as an unconditioned stimulus. Both before and after callosal lesion, bilateral CRs were the most frequent response pattern. Following callosal lesion, a statistically significant reduction in the percentage of CRs in at least one eye was observed in only two cats. In all seven animals, both before and after callosal lesion, the mean CR latencies of the ipsilateral eye were significantly shorter than those of the contralateral eye. Callosal lesion caused a significant increase in the mean CR latencies of both eyes in all subjects. These results provide evidence that the two hemispheres influence each other in controlling conditioned bilateral blinking by reciprocally exchanging facilitatory signals contributing to initiation of CRs in both eyes. PMID- 8721164 TI - Control of grip force during restraint of an object held between finger and thumb: responses of cutaneous afferents from the digits. AB - Unexpected pulling and pushing loads exerted by an object held with a precision grip evoke automatic and graded increases in the grip force (normal to the grip surfaces) that prevent escape of the object; unloading elicits a decrease in grip force. Anesthesia of the digital nerves has shown that these grip reactions depend on sensory signals from the digits. In the present study we assessed the capacity of tactile afferents from the digits to trigger and scale the evoked grip responses. Using tungsten microelectrodes inserted percutaneously into the median nerve of awake human subjects, unitary recordings were made from ten FA I and 13 FA II rapidly adapting afferents, and 12 SA I and 18 SA II slowly adapting afferents. While the subject held a manipulandum between a finger and the thumb, tangential load forces were applied to the receptor-bearing digit (index, middle, or ring finger or thumb) as trapezoidal load-force profiles with a plateau amplitude of 0.5-2.0 N and rates of loading and unloading at 2-8 N/s, or as "step loads" of 0.5 N delivered at 32 N/s. Such load trials were delivered in both the distal (pulling) and proximal (pushing) direction. FA I afferents responded consistently to the load forces, being recruited during the loading and unloading phases. During the loading ramp the ensemble discharge of the FA I afferents reflected the first time-derivative of the load force (i.e., the load-force rate). These afferents were relatively insensitive to the subject's grip force responses. However, high static finger forces appeared to suppress excitation of these afferents during the unloading phase. The FA II afferents were largely insensitive to the load trials: only with the step-loads did some afferents respond. Both classes of SA afferents were sensitive to load force and grip force, and discharge rates were graded by the rate of loading. The firing of the SA I afferents appeared to be relatively more influenced by the subject's grip force response than the discharge of the SA II afferents, which were more influenced by the load-force stimulus. The direction in which the tangential load force was applied to the skin influenced the firing of most afferents and in particular the SA II afferents. Individual afferents within each class (except for the FA IIs) responded to the loading ramp before the onset of the subject's grip response and may thus be responsible for initiating the automatic increase in grip force. However, nearly half of the FA I afferents recruited by the load trials responded to the loading phase early enough to trigger the subject's grip force response, whereas only ca. one-fifth of the SA Is and SA IIs did so. These observations, together with the high density of FA I receptors in the digits, might place the FA I afferents in a unique position to convey the information required to initiate and scale the reactive grip-force responses to the imposed load forces. PMID- 8721167 TI - Lip repair according to the principles of Delaire. What is its significance? AB - 45 patients with UCLP/UCL and BCLP/BCL had a lip repair at a mean age of 4 months according to the principles of Delaire from 1989 to 1993. Their aesthetic results and the influence upon growth and symmetry of the face was evaluated by standardized checking of possible negative points of the lip, nose and alveolus, by measuring different distances of the mid-face in standardized photographs and by comparing the data with the literature. The aesthetic results are equal to other studies and there are some indications of a better stimulation of growth and harmonization of the face. PMID- 8721168 TI - Laparoscopy for nonpalpable testis in childhood: is inguinal exploration necessary when vas and vessels are not seen? AB - A case of bilateral nonpalpable testes is described. Ultrasonography showed no testis. Laparoscopy revealed no intraabdominal testis, vas or vessels. Human chorionic gonadotrophine test suggested the presence of testicular tissue. Computed tomography showed a testis present in each inguinal canal. Inguinal exploration confirmed the finding. This case stresses the importance of careful interpretation of the laparoscopic findings together with other diagnostic methods in localizing testicular tissue in patients with nonpalpable testes. PMID- 8721169 TI - Extramucosal pyloromyotomy by laparoscopy. AB - Since 1990, we have performed extramucosal pyloromyotomies by laparoscopy in 70 infants presenting with congenital hypertrophic pyloric stenosis. Ever since the first cases, the technique and the instruments have been adapted. The surgical technique and the precautionary measures are here explained again. This technique is beneficial for infants with a better postoperative course and avoidance of any skin scar. PMID- 8721170 TI - Interest of umbilical fold incision for pyloromyotomy. AB - We report the results of a study concerning 80 infants undergoing a pyloromyotomy over a 3-and-a-half-year period. 40 infants had a right upper quadrant transverse rectus incision and 40 infants had an umbilical fold incision. Morbidity and cosmetic aspect of the two techniques were compared. We insist on the umbilical fold incision which presents the same advantages as the classic one and a better cosmetic result. PMID- 8721171 TI - Laparoscopic Swenson's procedure in children. AB - The technique is described, and the experience gained with 7 laparoscopic sigmoid resections is reported. The approach resembles Swenson's technique with only the preparatory part performed laparoscopically. With the most recent procedures, the anaesthesia time of the laparoscopic procedure did not exceed that of the open technique. There were no serious complications or blood losses. PMID- 8721172 TI - Gastroschisis in the decade of prenatal diagnosis: 1983-1993. AB - Fifty cases (24 males, 26 female) of gastroschisis were admitted to the Regional Neonatal Surgical Unit at the Royal Hospital for Sick Children in Yorkhill, Glasgow, between January 1983 and October 1993. The mean birthweight was 2.17 kgs and mean gestation was 35.8 weeks. Prenatal diagnosis was made by ultrasound scan in 39 cases (78%) and 33 (66%) were delivered in the Regional Obstetric Unit and Fetal Medicine Centre at the Queen Mother's Hospital, Yorkhill, Glasgow, immediately adjacent to the neonatal surgical unit. Spontaneous vaginal delivery occurred in 23 (46%) but Caesarean section was performed in 27 (54%). Sixteen Caesarean sections were emergencies and 11 elective (5 on account of dilated bowel loops). Associated bowel problems were noted in 11 (22%)--atresia (5), stenosis (2), ischaemia (1), duodenal perforation (1), ileal perforation (1) and jejunal band obstruction (1). Primary closure was achieved in 42 (84%) and there were 5 deaths (10%). Prenatal diagnosis and mode of delivery showed no direct correlation with mortality (10%) in this series. Emergency Caesarean section was necessary for fetal indications in 28%. The prediction of compromised or damaged intestine by prenatal ultrasound was unreliable and the majority of cases of atresia and stenosis escaped detection. However, it is concluded that prenatal diagnosis and delivery in a regional centre offers the best outcome for the fetus. Further study is needed to define "at risk" pregnancies and fetuses and to determine the role of intervention. PMID- 8721173 TI - National reevaluation of staging in Wilms tumor. AB - In order to evaluate if incorrect staging of Wilms tumor resulted in inadequate treatment a retrospective reevaluation was performed. During 1982-1990 153 patients were treated in Sweden. The review revealed that 6 cases were not Wilms tumor and 25 cases had incomplete information. The remaining material consisted of 122 cases. The survey of the charges revealed that the initial distribution was stage I 58, stage II 17, stage III 21, stage IV 15 and stage V 11. The stage was changed in 12 cases, mainly in the initial stage I. The causes for changing of the stage were thick needle puncture preoperatively in 4, capsule histologically not intact 4, tumor not radically excised 2, tumor rupture peroperatively 1 and lymph node malignancy 1. The staging procedure cannot be safely performed by a single doctor. It is dependent on cooperation in a pediatric oncologic team including the pediatric surgeon. PMID- 8721174 TI - Arthroscopy in the diagnostics and treatment of non-acute knee disorders in children. AB - 199 arthroscopies were performed on children with non-acute knee problems during 1985-1992 at the University Hospital of Oulu (76 boys and 123 girls). The mean age was 13 (2-15) years. The ratio between boys and girls was 1:1.6 showing the incidence among girls to be greater. The mean duration of symptoms was 16 (0.3 96) months before the arthroscopy. Major diagnostic groups were plica syndrome (43 cases), malalignment of the patella (29 cases), osteochondritis dissecans (28 cases) and arthritis of the knee (28 cases). The accuracy of clinical diagnosis was 47% in preadolescents and 51% in adolescents. Operations were performed arthroscopically in 71 cases, by open arthrotomy in 15 and by other extra articular operations in 39. These children were followed until no further treatment was needed. The mean follow-up time was 4 years and 6 months (20-97 months). A total of 172 (87%) patients needed no further treatment, while 27 (13%) children were rearthroscopied and a reoperation was performed on 15 of these. PMID- 8721175 TI - Age-related postoperative morphine requirements in children following major surgery--an assessment using patient-controlled analgesia (PCA). AB - To investigate if small children require less morphine for postoperative analgesia than do older children and adolescents we analysed the morphine consumption pattern of 28 consecutive children on intravenous patient-controlled analgesia (PCA) following major surgery. The median age-specific morphine requirements between 2 comparable groups of children aged 4-8 years and 9-15 years were compared. We used the Pharmacia-Deltec pump in all children and the same settings: a bolus dose of 25 microgram/kg, an 8 minutes lockout interval and no background infusion. In addition, all children received paracetamol as a supplemently to the morphine. In this study children aged 4-8 years had significantly higher total postoperative morphine requirements compared to children aged 9-15 years, i.e. 11.6 microgram/kg/hour and 7.5 microgram/kg/hour respectively (p = 0.037). Hence, we conclude that children of this age group may have a higher total postoperative morphine requirement following major surgery than older children and adolescents. PMID- 8721176 TI - Axillary involvement in pediatric Kikuchi's disease. AB - We report the first Israeli pediatric case of Kikuchi's disease in a 10-year-old male. He was hospitalized for evaluation of migrating arthralgia and fever. Axillary adenopathy was the main physical finding. A biopsy showed a histiocytic necrotizing lymphadenitis. Diagnosis, therapeutic options and the clinical significance of Kikuchi's disease are discussed. Kikuchi's disease should be considered in the differential diagnosis of fever of unknown origin in children, even in the absence of cervical adenopathy. A long follow-up is advised because of possible links with systemic lupus erythematosus and to detect eventual late recurrences. PMID- 8721177 TI - Inflammatory pseudotumor of the liver in children. A case report and review of the literature. AB - A case of hepatic inflammatory pseudotumor with preoperative diagnosis of malignancy in a 14-year-old girl is reported. The lesion manifested clinically with prolonged fever of unknown etiology. Accurate diagnosis was established by laparotomy, resection of the tumor and histological examination. The patient remains asymptomatic during a 2-year follow-up. Inflammatory pseudotumor of the liver should be considered in the differential diagnosis of such cases and the treatment might be more conservative after accurate diagnosis establishment, which is nevertheless difficult to achieve. PMID- 8721178 TI - Laparoscopic removal of Meckel's diverticula in children. AB - We report on 4 children where Meckel's diverticula (MD) were excised laparoscopically. In two of the children the finding of an MD was incidental and in two it was suspected preoperatively. In three of the patients the MD was ligated by means of an "endoloop" and could be removed in a similar manner to an appendix. In one patient the MD was resected by means of a stapler. No complications occurred. Subsequent histology confirmed the complete excision of the MD. Allowing for the size and shape of an MD, we prefer to excise it by means of an endoloop. An endoloop has the advantage that it can be used through a 5 mm trocar while a stapler has the disadvantage that it requires a larger 12 mm trocar and it leaves a foreign body, i.e. metal clips behind. PMID- 8721179 TI - Surgical reconstruction of a high cloacal anomaly with two short vaginas. AB - We treated a patient with a complex cloacal malformation: a high confluence cloaca and 2 separate short vaginas. At the age of 1 year and 2 months the rectocloacal fistula was closed because of frequent urinary infections, and the vaginas were separated from the wide urogenital sinus and anastomosed to the lower colon. At the age of 2 years and 4 months the lower colon, which had been anastomosed with the 2 vaginas, was pulled down to the perineum to serve as a lower vagina, and the sigmoid colon was pulled through to form a neorectum. She is now 7 years old and goes to school without problems of bowel function or urinary function except that catheterization is done twice a day. PMID- 8721180 TI - Megacalycosis on duplex system upper moiety. AB - The authors report a rare case of megacalycosis located on a duplex system upper moiety, and analyse the problems related to its pathology, etiopathogenesis, clinical manifestations and diagnosis. PMID- 8721181 TI - Chronic recurrent multifocal osteomyelitis (CRMO). AB - Pathogenesis of CRMO still remains unknown. Characteristic, but not pathognomonic for this syndrome are clinical course (age, sex, chronic recurrent, intermittent course), radiological findings (metaphyseal lesions), histology (chronic osteomyelitis without colliquation) and microbiological results (lack of pathogen, infectious agents). Favorable, self-limited long-term prognosis of CRMO has been assumed. Antibiotics provide no improvement. Course, severity and recurrency can be influenced positively by antiphlogistic substances, although this has not yet been proved. Whether surgical intervention beyond biopsies might cause improvement on the follow-up is unknown. Our experience (4 cases), and the literature demonstrate great clinical importance that unusual types of osteomyelitis (OM) can be within the differential diagnosis of multifocal osteolytic changes. PMID- 8721182 TI - Intrauterine intussusception due to Meckel's diverticulum as a cause of ileal atresia: analysis of 2 cases. PMID- 8721183 TI - Extent of pancreatic resection in the treatment of nesidioblastosis. PMID- 8721184 TI - [Identification and diagnosis of Taylorella equigenitalis by a DNA amplification method (PCR)]. AB - A polymerase chain reaction (PCR) for identification of Taylorella equigenitalis was developed. The oligonucleotide primers are based on the DNA sequence of the rrs gene of T. equigenitalis, encoding for the 16S ribosomal RNA. Analysis of 21 strains of T. equigenitalis from England, USA and Switzerland showed an amplification product of 410 bp with identical Sau3A restriction profile. The sensitivity of the PCR-Assay was estimated to detect 50 to 500 bacteria of T. equigenitalis in a mixture with frequently found contaminants. Further analysis of culture from 60 genital swabs, taken in the course of the control of the contagious equine metritis in horses and donkeys, of experimental assays as well as of two positive cases from the diagnostic showed that this PCR-assay can be used to identify and to detect strains of T. equigenitalis. In addition, preliminary results indicate that the method is also applicable for direct in vitro establishment of the presence of T. equigenitalis in clinical samples. PMID- 8721185 TI - [Typing of the apx toxin gene of Actinobacillus pleuropneumoniae using PCR]. AB - For the determination of the apx toxin genes in Actinobacillus pleuropneumoniae strains, five PCR reactions were developed which allow the detection of the activator and structural genes and the secretion genes of the toxins ApxI, ApxII and ApxIII. The oligonucleotide primers were chosen in order to amplify characteristic parts of the activator and structural genes apxICA, apxIICA and apxIIICA, and the secretion genes apxIBD and apxIIIBD. The annealing temperature of all five reaction was identical in order to allow the five reactions in a single PCR run. The differences in length between the individual amplified gene fragments allowed all product fragments to be separated in a single electrophoresis. This way, a typical toxin gene pattern could be obtained which is characteristic for the five toxin gene groups of A. pleuropneumoniae including serotypes 1, 5a, 5b, 9 and 11 (group 1), serotypes 2, 4, 6 and 8 (group 2), serotype 3 (group 3), serotypes 7 and 12 (group 4), and serotype 10 (group 5). The identification of the toxin genes, which have significance in virulence, enhance and facilitate differentiation of A. pleuropneumoniae and allow the detection of serotypes with an atypical toxin pattern. PMID- 8721186 TI - [Cloning of a protease gene family in Fasciola hepatica using the polymerase chain reaction (PCR)]. AB - RT-PCR and degenerative Oligonucleotide primers derived from conserved cysteine protease sequences were used to amplify seven different Fasciola hepatica cysteine protease cDNA clones (Fcp1-7). Five of the clones showed homology to proteases of the cathepsin L type, whereas two appeared to represent the cathepsin B family. The 5' and the 3' regions of Fcp1 were amplified using the rapid amplification of cDNA Ends PCR (RACE-PCR) protocol. The Fcp1 cDNA fragment was also subcloned in the expression vector pGEX and expressed as a glutathione-S transferase (GST) fusion protein. Antibodies, raised in rabbits against the GST:Fcp1 fusion protein, were used in Western blot analysis to examine expression in different life-cycle stages of F. hepatica. In extracts from immature and adult parasites proteins of 30 and 38 kDa were detected. In other stages, proteins of different molecular weight were recognized by anti-GST:Fcp1 antiserum, indicating stage-specific gene expression. PMID- 8721187 TI - [PCR for detection and characterization of parasites (Leishmania, Echinococcus, Microsporodia, Giardia)]. AB - The application of PCR for the diagnosis and the characterisation of parasites is discussed using four examples. (1) A PCR assay that was developed for the detection of Leishmania was shown to be as highly sensitive as in vitro cultivation of the parasites from lymph node aspirates and bone marrow biopsies. (2) Single eggs of Echinococcus multilocularis can principally be detected and identified by PCR. However, a cumbersome sample preparation is inevitable in order to remove PCR-inhibitory substances present in fox faeces. (3) Analysis of the amplified SSU rRNA gene of Encephalitozoon-like isolates from humans, rabbits and farm foxes confirmed for the first time that E. cuniculi-isolates from humans and animals are indistinguishable and that they are of importance as opportunistic parasites in HIV-infected patients. (4) Swiss isolates of Giardia originating from humans, calves, sheep and a dog could be allocated into three distinct genetic groups by analysing several PCR-amplified genes that encode for variant-specific surface proteins. There was no evidence for the presence of host specific genotypes. PMID- 8721188 TI - ["Arbitrarily primed PCR". Or: chance in the service of science]. AB - "Arbitrarily primed PCR" (AP-PCR) methods are based on the amplification of DNA with arbitrarily selected primers. In the present review article two selected applications of this methodical approach are summarized. Using the RAPD ("Random Amplified Polymorphic DNA") technique the genetic variability of different organisms can be analyzed by generating a genomic fingerprint. Echinococcus granulosus isolates (metacestodes) from Spain and Switzerland were comparatively characterized by the use of this molecular epidemiological tool. The following groups of genetically related isolates could be identified: (a) equine isolates (horse/donkey) from Spain and Switzerland, (b) cattle isolates from Switzerland, (c) goat isolates from Spain and pig isolates from both countries, (d) sheep, cattle and human isolates from Spain. Isolates derived from Swiss patients were forming a separate group with respect to their genetic relatedness. As a further application of the AP-PCR, the development of a novel technique for the construction of cDNA libraries from minute amounts of starting material using an Echinococcus multilocularis metacestode library as an example is presented. PMID- 8721189 TI - [Diagnostic PCR in veterinary parasitology: tritrichomonosis, neosporosis/toxoplasmosis echinococcosis/cysticercosis]. AB - The PCR is now widely introduced as a diagnostic and epidemiological tool in veterinary and human parasitology. Certain parasitic infections are detected using PCR with much higher sensitivity compared to conventional methods, and novel molecular approaches are considered for the analysis of infectiological questions. A broad spectrum of parasites relevant for veterinary and human parasitology already can be diagnosed by PCR. In the present review article, PCR based methods developped or applied at the Institute of Parasitology in Berne for the detection of protozoan infections (Tritrichomonosis, Neosporosis, Toxoplasmosis) and helminthic infections (Echinococcosis, Taeniosis) are summarized. PMID- 8721190 TI - [Analysis of microsatellite polymorphisms in Bernese mountain dogs and Newfoundlands]. AB - As the number of detected inherited diseases in dogs is increasing, linkage mapping offers an opportunity for the localisation of the responsible genes. In humans microsatellites are ideal markers for constructing high-resolution genetic maps. Twenty-five dog specific microsatellites which are highly polymorphic in mixed dog breeds, were tested by PCR in a group of unrelated Newfoundlands and Bernese Mountain dogs. The polymorphisms found in this two breeds were clearly lower than in the mixed breeds suggesting that only 40% of the markers tested would be useful for linkage analysis. In addition the allele frequencies and distributions differed among this two purebred but related breeds. Our data suggest that a selection for useful markers for each specific breed has to be performed prior to linkage analysis. PMID- 8721191 TI - [Characterization of inbred strains of rats by microsatellite analysis]. AB - Microsatellite analysis was applied to identify the inbred rat strains LEW, SHR and WKY. Of a set of seven commercially available primer pairs three pairs were used individually and two times two in duplex polymerase chain reactions (PCR). The products were analyzed in ethidium bromide stained agarose gels. The resulting banding patterns clearly demonstrated the homogeneity within strains and the differences between strains. PMID- 8721192 TI - [PCR--the step to conclusive paternity in parentage control cases in cattle]. AB - PCR-based technology used for genotyping microsatellites is going to add new dimensions to parentage control in cattle. In the very near future the determination of microsatellite polymorphisms is expected to replace the former parentage control of conventional blood typing. Parentage control by microsatellite analysis is even more reliable than by the routine haemogenetic procedures. The potential of microsatellite polymorphisms to clarify family relationships is illustrated by 100 cases which could not be solved by conventional blood typing. PMID- 8721194 TI - [The division of mental suffering -- which form of the therapy: an ethnoanthropological contribution]. PMID- 8721193 TI - [Isolation of Y-specific probes in cattle applying genetically representational difference analysis]. AB - Genetically directed representational difference analysis (GDRDA) allows to isolate DNA fragments that differ between complex genomes. This approach was applied in the present work to isolate Y-specific bovine sequences. Of eight isolated and cloned fragments, two proved to be Y-specific. PMID- 8721195 TI - [The family burden in chronic diseases. The study of validity and reproducibility of the questionnaire on the impact of the chronic disease on the family (IMPAT)]. AB - OBJECTIVE: The "Questionario sull'Impatto della Malattia Cronica sulla Famiglia" (IMPAT) ("Impact-on-Family Scale", by Stein & Reissman) in its Italian version (by Casari & Fantino, modified) has been used to evaluate family burden in relatives of chronic patients with cancer, chronic patients with other internistic pathology and acute patients. The aims of the study were to evaluate validity and reliability of the Impat and to compare family burden and anxoius/depressive symptoms in the three groups of relatives. METHOD: 35 relatives of cancer patients, 20 relatives of patients with chronic, non neoplastic illness and 20 relatives of acute patients were studied. 85% of relatives of cancer patients was assisted continuously by volunteers and psychiatrists with regard to their psychological problems. Impat questionnaire, Beck Depression Inventory and State-Trait Anxiety Inventory were used for psychological evaluation. RESULTS: Validity of the Impat was confirmed. Mean scores from relatives of chronic patients were higher than scores from relatives of acute patients (ANOVA, f = 38.0; df = 2,72; p < 0.0001). Test-retest analysis was satisfactory (Person's test, N = 20 r = 0.77, p < 0.0001). Both groups of relatives of chronic patients showed higher level of anxious-depressive symptoms compared to relatives of acute subjects. No differences were found between relatives of neoplastic and non neoplastic chronic patients. CONCLUSIONS: The italian version of Impat has been confirmed as a useful and simple means of evaluation of family burden in this context. Psychosocial support seems to reduce distress in relatives of cancer patients. PMID- 8721196 TI - [Stress, depression and anxiety in patients with headache. Considerations of literature and experimental data]. AB - After having examined the main studies which suggest the existence of correlations between anxiety, depression and cephalagia, the authors report the results of a study performed in a sample population of some three hundred subjects. During the presentation and discussion of results they also comment on some of the main psychodynamic theories relating to the etiology of idiopathic cephalea. In particular, they examine the theories which relate the onset of cephalea to the inhibition of thought and those which instead connect the onset of cephalea to aggressiveness. The present study also comments on possible interpretative models along Individual-psychology lines concerning stress-related somatisation. PMID- 8721197 TI - [Considerations on patient admission to the psychiatric services]. AB - In this article the authors consider in a psychoanalytic way the theme of the reception inside a psychiatric territorial service. At the beginning, they underline some characteristics about the psychiatric demand and the setting in a public context, then they describe their experience. After examining how the initial time of the relation influences the subsequent evolution of the therapeutic relation, they conclude with the individuation in the style of relation showed by the patient the lived field, that can be changed into a mental and representative space from an acted and fragmentary space. The skill of the service in changing the field of treatment proposed by the patient, in a field which transforms the action into therapeutic trials, becomes essential at the beginning of the relationship. PMID- 8721198 TI - [The neuropsychological study of 21 patients with expanding cerebral lesions. Preliminary results]. AB - Twenty-one right-handed patients with cerebral expanding space-occupying lesion underwent neuropsychological evaluation before surgery. Testing included the Bender Motor Gestalt Test (BMGT), the Benton Visual Retention Test (BVRT), and four subtests of the Wechsler Adult Intelligence Scale (WAIS), namely Digit span, Digit symbol, Picture completion, and Block design. Statistical analysis showed significant differences in cognitive efficiency between the groups defined by the side and biological behaviour of the lesions. PMID- 8721199 TI - [Effects of psychopathological components of the onset of Alzheimer's disease]. AB - A particular importance can assume, especially during the onset of Alzheimer's disease, the psychiatric symptomatology characterized by depressive mood, auto and hetero-aggressive behaviour, psychomotor agitation, anxiety disorders, sleep disorders, delusions and hallucinations, these pictures showing a prevalence of 30%. 13 patients affected by Alzheimer's disease at the onset, of both sexes, age ranging from 45 to 80 years, were included in the study. The clinical picture was assessed by BDS, GDS, ICS, IADL, BIMC, MMSE, ADAS, a neuropsychological test battery such as Token Test, verbal fluency test for semantic categories, prose memory test, scribble discrimination test, numeric matrix test, Raven test, judgement of line orientation. A computerized test battery by NYU Computerized Test Battery have been also administered. The psychiatric clinical picture has been evaluated by HRS-D and the non cognitive session of ADAS. There was a positive significant (r = 0.85, p = 0.0004) correlation between depressive symptomatology evaluated at HRS-D and deterioration of daily living activities evaluated at the Blessed Dementia Scale (BDS). On the other hand the deterioration of cognitive capacities was not significantly correlated with a worsening of behavioural aspects. In conclusion seems to have a particular importance, at the onset of Alzheimer's disease, the psychopathological component that often seems to be the principal component in the determination of the early deterioration of daily living and behaviour or patients. PMID- 8721200 TI - [Sexual dysfunction in men and epilepsy]. AB - Sexual dysfunctions may represent a symptomatological aspect of epilepsy or it may be a consequence of the pharmacological therapy. This kind of trouble is frequently associated to primary generalized epilepsy and focal secondary epilepsy. In our investigation we considered a group of 30 males during treatment with drugs like phenobarbital, carbamazepine and valproate, at the "Center for the Study and the Treatment of Epilepsy", University of Turin, using a sexological and psychosomatic questionnaire. The purpose was the analysis of the sexuality in relation to a specific kind of epilepsy or to a particular drug. PMID- 8721201 TI - [Pain in psychopathology. Brief phenomenological analysis]. AB - The problem of pain is felt more and more by people at various levels of privacy and social life. It is important to examine the possibility of pain as "communicative metalanguage", or language of languages. This language is deeply analogic, metaphoric, symbolic, experienced. The authors, in this paper, try to consider the problem of pain from a psychopathologic point of view and outline some lines of phenomenological analysis, from depression to schizophrenia. PMID- 8721202 TI - [Root canal overfilling. Periapical immune reactions]. AB - The authors studied the immunological answer of periapex to look for causes of osteolysis. They did six apicoectomies in teeth with periapical lesions containing extruded material, and found an inflammatory infiltrate of T lymphocytes and macrophages with birifrangent bodies. They suppose that this immunitary answer to extruded material, prevents the periapex cure. PMID- 8721203 TI - Butyric acid, a metabolic end product of anaerobic bacteria, inhibits B lymphocyte function. AB - The effect on B-lymphocytes of butyric acid, a metabolic by-product formed by selected anaerobic bacteria of the subgingival plaque and present in mM amounts in the crevicular fluid of patients affected by adult periodontitis was investigated. A dose-dependent inhibition of both mitogenesis and Ig production in B-cells challenged with formalinized Staphylococcus aureus, strain Cowan I (SAC), was observed. A role of this molecule in the downregulation of the local immune response in periodontal patients is proposed. PMID- 8721204 TI - [Scanning electron microscopic study of the surface finish of the round sections of preformed arches of NiTi (orthodontic wires made of nickel/titanium]. AB - The authors indicate the advantageous characteristics of NiTi wire, focusing the attention on finishing surface accuracy. Every surface damage can become the start of a fracture. PMID- 8721205 TI - [Indications for mandibular periosteotomy in the treatment of cancer of the oral cavity]. AB - Marginal mandibulectomy is a compromise between preserving the mandibular structure and radical oncological surgery. In order to evaluate the correctness of the indications for this operation, a group of 47 patients with oral carcinoma underwent direct surgery or associated chemoradiotherapy and surgery. Surgery was programmed on the basis of clinical and radiological examinations and the specific evaluation of the distance of the tumour mass from the mandibular surface. This approach was justified by the lack of correlation, in anatomic and clinical terms, between tumoral infiltration of the mandible and metastatic recurrence in the cervical lymph nodes. Lastly, the varying pattern of tumoral invasion of the various mandibular regions (thickness of cortical bone, presence of foramen) induced the authors to distinguish the indications for conservative treatment by segments. According to international literature and on the basis of the authors' past experience, marginal mandibulectomy represents an extremely interesting method since it associates a satisfactory aesthetic-functional level with a radical oncological approach, in particular with regard to the symphyseal and parasymphyseal region owing its structural characteristics and reconstructive importance. The indications for this type of surgery are therefore in theory based primarily on oncological aspects, and subsequently on the various anatomic and reconstructive aspects of the various mandibular segments. PMID- 8721206 TI - [Recurrent aphthous stomatitis: current etiopathogenetic and therapeutic concepts]. AB - The authors review the literature on aetiopathogenesis and therapeutic management of recurrent aphthous stomatitis. The data regarding the role of genetic, nutritional and microbiological factors in the genesis of recurrent aphthous stomatitis has been particularly examined. Despite significant associations with some antigens HLA have been reported in Southern Europe, there is no clear genetic predisposition in recurrent aphthous stomatitis. Several studies have analyzed the importance of iron, folic acid and vitamin B12 deficiencies, gluten intolerance and sensitivity to certain foods in the triggering of recurrent aphthous stomatitis however the results have been controversial. Recently, it has been suggested that recurrent aphthous stomatitis could be caused by reactivation of varicella-zoster virus and/or cytomegalovirus but these viruses may be reactivated by the immunodysregulation known to underlie recurrent aphthous stomatitis. Moreover, antiviral drugs appear to have only an equivocal effect on recurrent aphthous stomatitis. Recurrent aphthous stomatitis is probably determined by immunological mechanisms although there actually no unifying hypothesis which attempt to integrate the results of the many immunologic studies on recurrent aphotous stomatitis. Moreover, the target antigen and the cause of recurrences of recurrent aphthous stomatitis are still unknown. As far as the management of this disease it is important to recognize recurrent aphthous stomatitis secondary to systemic diseases like Behcet's syndrome, gluten enteropathy and haematinics deficiencies. Subsequently, the symptoms can be reduced with several drugs (mainly topical corticosteroids) but there are no effective therapies preventing recurrences. PMID- 8721207 TI - [Therapeutic management of oral lichen planus. Presentation of a representative case]. AB - The authors describe the more efficient therapeutical plans of oral lichen planus, reducing the tendency to relapse. PMID- 8721209 TI - [Corono-radicular amputation of mandibular molars: indication, technic and prosthetic utilization. Apropos of a clinical case]. AB - Lesions to the radicular bifurcation generally entail the removal of the tooth in question. This leads to the need to create a prosthetic bridge to compensate for the absence of the extracted tooth. This solution is not readily accepted by the patient who wishes to avoid involving other teeth. In these cases rhizectomy and the execution of an inlay on an adjacent tooth may satisfy the patient and resolve the clinical symptoms. The authors report a case of rhizectomy and prosthetic reconstruction of a tooth using inlay. PMID- 8721208 TI - [Post-traumatic myo-arthropathy of the mandibular joint. Report of a case]. AB - The author reports a case of post-traumatic myoarthropathy which was resolved through the search for a new centric ratio which was subsequently transformed in ICP with a varied vertical dimension. This was based on the assumption that post traumatic intracapsular modifications impose this therapeutic objective given that they do not fall within the range of muscular adaptability. PMID- 8721210 TI - [Maxillofacial-dental anomalies in cleidocranial dysostosis]. AB - The authors evaluated the dento-maxillofacial anomalies in cleido-cranial dysplasia; the use of a removable orthodontic appliance can solve orthodontic problems particularly the deficiency of anchorage. PMID- 8721211 TI - A minimum essential structure of LN-3 elicitor activity in bean cotyledons. AB - A great deal of an elicitor-active oligosaccharide (1.47 g) was obtained from 40 g of polysaccharide laminaran with the guidance of elicitor activity in the bean cotyledon assay. Physicochemical means suggested that the oligosaccharide was a beta-1,3- and beta-1,6-linked triglucoside. This was a partial structure of LN-3, having elicitor activity to alfalfa and bean. The triglucoside exhibited a specific elicitor activity to bean, not to alfalfa and pea. The highly digested hydrolysate containing monomer and dimers as major components did not show elicitor activity in the bean cotyledon assay. The structural feature was essential for elicitation of phytoalexin accumulation in bean cotyledons. These results suggested that this triglucoside had a specific elicitor activity and a minimum elicitor-active entity to bean cotyledons. PMID- 8721212 TI - Isolation, characterization, and substrate specificity of the plasma membrane ATPase of the hlophilic achaeon Haloferax volcanii. AB - Isolated membranes of the moderate halophilic bacterium Haloferax volcanii are able to hydrolyze ATP via an ATPase, which needs the presence of Mg2+ or Mn2+, high concentrations of NaCl, a pH value of 9, and high temperatures with an optimum at 60 degrees C. We have not found any phosphatase activity in the preparations. We developed a purification method for the isolated enzyme with an enrichment factor of 90. SDS-gel electrophoresis of the partially purified enzyme of Haloferax volcanii showed putative ATPase subunits of 63, 51, 37, and 12 kDa. N-ethylmaleimide (NEM) a specific inhibitor for V-ATPases, which alkylates cyteines, inhibited the enzyme slightly. Binding of tritiated NEM to the isolated ATPase fractions resulted in labelling of the 63 and 51 kDa peptides. Using PCR with degenerate oligonucleotides, we could clone and sequence a gene cluster encoding the A1 part of the halophilic ATPase. The described genes are organized in an operon in the order D, C, E, B, A, named alphabetically according to their decreasing size. The deduced products of 64.5, 52, 38.7, 22, and 11.6 kDa confirm the results of the partial purification of the ATPase. Biochemical characterization of the Haloferax volcanii ATPase gave the following results: In presence of Mn2+ higher rates of ATP hydrolysis could be observed than in presence of Mg2+, but free manganese ions inhibited the enzyme activity of the ATPase. Calculation of the true concentrations of the complex between ATP and the respective divalent metal ion led to determination of Michaelis-Menten constants for ATP in the hydrolysis direction of 1 mM in presence of MgCl2 and 0.24 mM in presence of MnCl2. Sodium chloride concentrations in the molar range induce changes in KM by a factor of about 10. The enzyme is specific for ATP; other nucleotides including GTP and ADP are competitive inhibitors of ATP hydrolysis. PMID- 8721213 TI - Toxicity of Palicourea marcgravii: combined effects of fluoroacetate, N methyltyramine and 2-methyltetrahydro-beta-carboline. AB - Feeding experiments carried out with cattle and horses could prove the toxic effects of P. marcgravii (Rubiaceae) in all cases. The typical symptoms of "sudden death", however, are observed in ruminants only. This difference could not be explained so far. Apart from fluoroacetate, two more substances also have influence the toxic effects and have been isolated from P. marcgravii for the first time: N-methyltyramine and 2-methyltetrahydro-beta-carboline (2-Me THBC). Structure elucidation of these compounds is mainly accomplished by 1H-NMR, 13C NMR and MS techniques. Due to the small quantity of fluoroacetate (5.4 micrograms/g plant), the main toxic effect obviously lies in the two discovered substances. In contrast to the slow death of horses (monogastriers), the "sudden death syndrome" of cattle (ruminants) can be explained as a result of the higher resorbility of these two substances in the gastro-intestinal system. Given orally, both substances influence the monoamine oxidase type A (MAO-A): N methyltyramine acts as a competitive substrate, and 2-Me THBC is one of the most effective MAO-A-inhibitors. Thus, the decomposition of the specific MAO-A substrates noradrenaline and adrenaline as well as of N-methyltyramine itself is inhibited. The alpha- and beta-receptors of the sympathetic system are stimulated more strongly, which leads to a drastic rise in blood pressure and thereby to a more rapid distribution of fluoroacetate in the body. This results in a reinforced input of fluoroacetate in the cells of especially active organs of the body (heart etc.). Thus, even smaller quantities of fluoroacetate are lethal. PMID- 8721214 TI - Effect of Mg(2+)-ATP on acetylcholinesterase of Electrophorus electricus (L.). AB - The effect of Mg(2+)-ATP on purified acetylcholinesterase (AChE) from electric tissue of Electrophorus electricus (L.) was studied. The enzymatic activities were measured with acetylcholine and acetylthiocholine as substrates. The kinetic parameters Vmax, Km and Hill coefficient (nH), for acetylcholine and acetylthiocholine were modified with Mg(2+)-ATP. It was shown that acetylcholinesterase presents an apparent activation at high concentration of substrates and an inhibition in the presence of Mg(2+)-ATP at low concentration of acetylcholine and acetylthiocholine. In addition, the data suggest that Mg(2+) ATP induced an allosteric modulation of the acetylcholinesterase obtained from Electrophorous electricus (L.), and indicate an active adenosine triphosphate participation during cholinergic activity. PMID- 8721215 TI - The influence of alkoxymethyl purine and pyrimidine acyclonucleosides on growth inhibition of Kirkman-Robbins hepatoma and possible mechanism of their cytostatic activity. AB - Newly synthesized allyloxymethyl purine and pyrimidine acyclonucleosides [Fig. 1, comp. 1-6] were tested in Syrian hamster, six days after heterotransplantations of Kirkman-Robbins hepatoma and compared with Th5, Th5P and PMT [Fig. 1, comp. 7 9]. 48 hours after intraperitoneal (i.p.) administration of AMT and Th5 in a dose of 80 mg per kg body weight, these compounds reduced tumor weight by 42%, while AMU (in the same dose) by 30%. The inhibition of tumor weight is accompanied by a decrease in dThd and dGuo kinase activities in tumor cytosol by AMU (36% and 33%, respectively) by AMT (59% and 53%, respectively) and by Th5 (58% and 55%, respectively). AMU, AMT and Th5 are phosphorylated in vivo by kinases present in cytosol of growing hepatoma to mono, di and triphosphates, but allyloxymethyl residue of AMU and AMT is first hydrated to hydroxypropoxymethyl residue, having CH2OH group. The lack of phosphorylation of PMT in vivo (having saturated propoxymethyl residue) and phosphorylation of Th5P (when used as a substrate for dNMP kinase) only to Th5 diphosphate suggested that AMU, AMT and Th5 triphosphates are responsible for the inhibition of dTMP and dGMP synthesis. PMID- 8721216 TI - Evidence of surface diffusion of water molecules on proteins of rabbit lens by 1H NMR relaxation measurements. AB - In this work, we propose a relaxation model for the interpretation of NMR proton spinlattice and spin-spin relaxation times of mammalian lenses. The framework for this model is based on nuclear magnetic spin-lattice relaxation measurements as a function of temperature at different Larmor frequencies for whole rabbit lenses and fragments of the lens. According to this model, two different dynamic processes of the water molecules determine the relaxation behaviour, namely rotational diffusion and translational surface diffusion. These dynamic processes in conjunction with a two site exchange model give a good explanation of all the measured relaxation data. From the experimental data, we were able to obtain the activation parameters for rotational and translational diffusion of bound lens water. Correlation times of 2.1 x 10(-11) sec and 2.5 x 10(-9) sec and activation energies of 20.5 kJ/mol and 22.5 kJ/mol respectively were found at 308K. At low Larmor frequencies (< or = 100 MHz) the longitudinal relaxation is mainly determined by translational surface diffusion of bound water with a mean square displacement of 1.5 nm, whereas at higher frequencies (> or = 300 MHz), rotational diffusion is the main relaxation mechanism. The spin-spin relaxation is determined by translational diffusion over the whole frequency range and therefore shows only a very small dispersion. By our model it is possible to explain: 1) the strikingly large difference between the T1 value and the T2A and T2B values observed in the lens and 2) the different values of the activation energies measured at different fields for the lens. PMID- 8721217 TI - The response of rat serum lipids to diets of varying composition or contaminated with organochlorine pesticides. AB - The effects of different diets (high carbohydrate, high protein, high fat) and diets contaminated with polychlorinated biphenyls (PCBs) and/or gamma hexachlorocyclohexane (lindane) on the levels of serum triglycerides, cholesterol and phospholipids were investigated in Wistar rats. Serum triglyceride levels differed significantly among the diets, while those of cholesterol and phospholipids were much less affected by the diet composition. A change in diet composition resulted in a gradual adaptation to the lipid levels characteristic of the new diet with major variations including oscillations. There was, however, no specific component of a diet that could be associated with any specific change in serum lipids. While feed deprivation decreased the serum lipids (40-65% in 3 days), refeeding the starved animals caused pronounced increases of the lipids that were different among the diets. The response of the triglyceride levels was the strongest (up to 10 times the starvation levels) followed by those of the phospholipids (4-fold) and cholesterol (2.5-fold). Response of the triglyceride levels peaked within 1 or 2 days of refeeding, whereas those of cholesterol and phospholipids took 4 days to reach the maximum. Feeding PCB-contaminated diets increased the serum lipids in a dose-dependent manner (15-250 ppm). Higher PCB concentrations were increasingly inhibitory (350 ppm) or overtly toxic (> 400 ppm). Elevated lipids returned to the starting levels immediately after peaking (triglycerides) or only after several days (cholesterol, phospholipids) but with an earlier onset at lower PCB concentrations. Refeeding starved animals with PCB contaminated diets also increased the serum lipids dose-dependently. Feeding lindane-containing diets (50-150 ppm) as well as refeeding animals with lindane diets resulted in a considerable increase of the triglyceride levels, while cholesterol and phospholipids increased much less. Higher lindane concentrations (250 ppm) were inhibitory. The outcome on serum lipid levels on feeding diets contaminated with both PCBs and lindane was basically additive. PMID- 8721218 TI - The light-stimulated cytosolic calcium transient in Limulus ventral nerve photoreceptors: two components in the rising phase. AB - 1. Light-evoked electrical responses were measured in Limulus ventral nerve photoreceptors simultaneously with changes in the cytosolic free calcium concentration, by means of arsenazo III. 2. It has been shown here for the first time that the rise of the arsenazo signal consists of two phases. Only the slow phase in the rise of the signal depends on the membrane voltage. The reversal potential of the amplitude of this slow rising phase was about +196 mV. After removal of external calcium the reversal potential was about +20 mV. 3. When Na+ in the superfusate was replaced by Li+, the amplitude of the fast rising phase was reduced on the average to 50%, while the slow rising phase was not affected. 4. We conclude that the fast rising phase is caused by release of calcium from internal stores while the slow increase in [Ca2+]i is due to influx across the plasma membrane, possibly through light-activated ion channels. PMID- 8721219 TI - Effect of 2'-deoxyguanosine oxidation at C8 position on N-glycosidic bond stability. AB - The influence of 2'-deoxyguanosine (dG) oxidation at the C-8 position on N glycosidic bond stability was investigated. A kinetic analysis of dG and 8-oxo-2' deoxyguanosine (8-oxodG) depurination reactions was carried out in water solutions at pH ranging from 2 to 7.4 and temperature of 100 degrees C. The results indicate that N-glycosidic bond of 8-oxodG is significantly more stable in comparison with dG at any pH applied. At pH 5.1 hydrolysis rate of dG is 4.5 fold higher than that for 8-oxodG. The chemical stability of the modified nucleoside in oxidatively damaged DNA is one of important factors contributing to its mutagenic potential. Results of our experiments indicate that 8-oxodG, potentially mutagenic and carcinogenic nucleoside, is hardly susceptible to spontaneous depurination and its removal from cellular DNA depends mostly on the activity of DNA repair enzymes. PMID- 8721220 TI - Thermal gelation of meat batters as a function of type and level of fat and protein content. AB - The rheological behaviour of meat batters during heating was analysed as a function of protein level (10-16%), type (pork back or perirenal/peritoneal fat) and amount (10-22%) of fat used. Fat thermal behaviour was studied by differential scanning calorimetry and rheological properties of batters were assessed using non-destructive measurements (thermal scanning rigidity monitor). The higher the protein content, the higher were the rigidity values displayed by the batters, irrespective of fat type, although the magnitude of these values appeared to be dependent on the amount and characteristics of the fat. The lower the fat content, the lower were the rigidity values of the batters. This behaviour pattern was influenced by the amount of protein present. In general, samples containing perirenal/peritoneal fat exhibited lower rigidity values at high temperatures, whereas at less than 35-40 degrees C, the opposite appeared to be the case. PMID- 8721221 TI - Migration of dioctyladipate plasticizer from food-grade PVC film into chicken meat products: effect of gamma-radiation. AB - Food-grade PVC film containing 28.3% dioctyladipate (DOA) plasticizer was used to wrap chicken meat samples, with and without skin, contained in a polystyrene tray. Samples were then irradiated with gamma-radiation [60Co] at doses equal to 4 kGy and 9 kGy corresponding to "cold pasteurization". Irraddiation was carried out at 8-10 degrees C and samples were subsequently stored at 4-5 degrees C. Contaminated chicken meat samples were analysed for DOA at intervals between 7 h and 240 h of contact, using an indirect GC method. Identical non-irradiated (control) samples were also analysed for their DOA content. Results showed no statistically significant differences in migrated amounts of DOA between irradiated and non-irradiated samples. Neither were differences observed between samples irradiated at 4 kGy and 9 kGy. This was supported by identical IR spectra recorded for irradiated and non-irradiated samples and leads to the conclusion that, at such intermediate radiation doses ( < or = kGy), the migration characteristics of PVC film are not affected. DOA migration was found to be time dependent, approaching equilibrium after approximately 170 h for the chicken flesh plus skin samples and 120 h for the chicken flesh samples. The amount of DOA migrated into chicken flesh plus skin samples was significantly greater (3.2 22.3 mg/dm2) than that for chicken flesh samples (0.9-8.9 mg/dm2). After 240 h of sample/film contact under refrigeration, loss of DOA was approximately 35.6% for chicken flesh plus skin samples and 14.3% for chicken flesh samples. Sample spoilage, as demonstrated by off-odour development, occurred after approximately 120 h of refrigerated storage. Diffusion coefficients for DOA were calculated and were found to be lower for chicken flesh (1 x 10(-13) than for flesh plus skin (4.4 x 10(-13)) samples. PMID- 8721222 TI - [Characterization of low molecular weight zinc species in normal commercial vegetable foodstuffs]. AB - In six different ordinary vegetables, namely kohlrabi, Chinese cabbage, chard, leak, spinach and Jerusalem artichoke, zinc was mainly found as low molecular weight species. In the present study, these important zinc compounds are further investigated. The determinations of the metal are performed by ET-AAS. The zinc complexes of all vegetables are anionic at pH 8.0 and show similar elution behaviour in gel permeation and anion exchange chromatography. Consequently, a great resemblance in structure between the low molecular weight zinc species from the different vegetables can be supposed. Exemplary, the zinc complexes of kohlrabi and Chinese cabbage are further examined. In more purified samples of these vegetables compared to zinc neither stoichiometric amounts of free protein amino acids nor nicotianamine, free malic acid, citric acid or phytic acid have been detected. Mainly glutamic acid is found in molar excess to zinc after acid hydrolysis in both cases. The cysteine contents of both zinc-binding fractions are very low. Conclusively, the well-known gamma-glutamylcysteinyl-glycines (phytochelatines) can not be responsible for the bonding of zinc in both ordinary vegetables. We suppose that zinc in kohlrabi and Chinese cabbage is bound to a glutamic acid derivative unknown as yet, possibly a malic acid ester. PMID- 8721223 TI - New records of Tabanidae (Diptera) from Saudi Arabia, first record of Atylotus venturii Leclercq and Tabanus separatus Efflatoun. AB - Atylotus venturii Leclercq and Tabanus separatu Efflatoun (Diptera: Tabanidae) are 2 additional species in Saudi Arabia; they lift up to 26 taxa known from the country. Some comments on new record of other species, zoogeographical distribution of Tabanus sufis, taxonomic significance of eye pattern of Tabanidae, exemplification with T. albifacies and T. unifasciatus, rehydration of dry specimens, are presented. PMID- 8721224 TI - Detection of Trichinella pseudospiralis antigen in the parenteral phase in experimentally infected mice. AB - Antigen distribution of T. pseudospiralis in muscles was studied at 10 days, 2,3 and 5 weeks post infection. Immunocytolocalization of antigen in the muscle was performed by peroxidase antiperoxidase (PAP) and immunogold silver staining techniques. Weak positive reaction of the stichosome and the cuticle was observed at 10 days post-infection which increased to attain its highest level in the fully infective stage larvae 3 and 5 weeks post infection. The intensity of staining reaction in the cytoplasm and nucleoplasm of infected host cell and surrounding tissue was highest at 5 weeks p.i. The small nuclei of the host cells and the inflammatory cells were unstained. PMID- 8721225 TI - Immunization against Trichinella spiralis using antigens from different life cycle stages experimental study in mice. AB - Crude antigen preparations from different life-cycle stages of T. spiralis using 10 micrograms and 100 micrograms antigen/dose, were tested for their ability to immunize mice against homologous challenge infection. Immunization was assessed by accelerated expulsion of adult worms, reduction in female worm fecundity and decreased yield of muscle larvae. The dose of 100 micrograms proved to be more protective in all antigen preparations. The AA provided highly significant protection against adult stages (89%) and muscle larvae (80%). NLA produced highly significant protection against muscle larvae only (76%). MLA provided highly significant protection against adult stages (74%), female fecundity (74%) and muscle larvae (80%). Combination of AA and MLA produced highly significant protection against adult stages (96%), female fecundity (73%) and muscle larvae (86). The results show that all antigen preparations provided variable, considerable but partial protection against challenge infection. The variations in immunization potential are discussed. PMID- 8721226 TI - Effect of growth hormone administration on serum somatomedin-C levels in bilharzial and liver disorders patients. AB - This work was designed in an attempt to clarify the effect of growth hormone injection on the serum somatomedin-C (SM-C) levels in Egyptian children with (1) urinary bilharziasis, (2) advanced bilharzial hepatic fibrosis, and (3) children suffering from chronic hepatitis, in comparing with 11 healthy age- and sex matched children,, served as controls. SM-C levels were studied prior and after injection of human growth hormone (hGH). The basal and hGH-stimulated SM-C levels were significantly reduced in bilharzial patients compared with controls. Patients with hepatitis had significantly lower serum SM-C values prior or post hGH administration. Liver tests carried out for bilharzial patients showed impaired function both on admission and after treatment. It can concluded from this work that shortness in children with bilharzial hepatic fibrosis may be result of hepatic and endocrinal factors and nutritional. Also, our results suggested that the delayed skeletal maturation in chronic hepatitis cases is probably secondary to liver dysfunction, malnutrition and associated endocrinopathies. PMID- 8721227 TI - Flea ectoparasites of commensal rodents in Shebin El Kom, Menoufia Governorate, Egypt. AB - Rodents population are widely distributed in some Egyptian Governorates, particularly those in the Nile Delta. Generally speaking, rodents particularly those living in close association with man, play a major role in human health, welfare and economy. Besides, their arthropod ectoparasites are important vectors of pathogenic agents as well as they themselves are causative agents of many allergic disorders. No doubt, the increase in rodent population is followed by an increase in many zoonotic diseases. In Shebin El Kom city, the capital of Menoufia Governorate three commensal rodents were identified. They were Mus musculus (house mouse), Rattus rattus (black rat) and Rattus norvegicus (Norway rat). The maximal rat index was in Autumn (0.69) and minimal was in Winter (0.2). Five species of fleas were collected from the rodents. They were Xenopsylla cheopis (57.7%), Xenopsylla ramesis. (12.7%), Pulex irritans (12.7%), Leptopsylla segnis (11.3%) and Ctenocephalides felis (5.6%). The maximal flea index was in Spring (1.02) and the minimal was in Winter (zero) The highly infested rodent species was R. rattus (0.65/rat), R. norvegicus (0.62/rat) and the lowest was M. musculus (0.45/mouse). The overall flea index was 0.59. The role of fleas in disease transmission was included. No doubt, to carry out feasible control measures against rodents and their ectoparasites complete understanding of their bionomics must be well studied. PMID- 8721228 TI - Tumour necrosis factor-alpha (TNF-alpha) in human toxoplasmosis. AB - Tumour necrosis factor alpha (TNF-alpha) is a cytokine that is secreted by activated macrophages and monocytes. This work studied TNF-alpha level in sera of patients with toxoplasmosis. Twenty eight female patients (12 of whom had obstetric troubles) and 17 healthy controls were the subjects in this work. According to IHA test and detection of IgM antibodies, three groups of patients were defined, group (I) patients with acute infection, group (II) patients with chronic infection and had high antibody titre, group (III) patients with chronic infections and had low antibody titre. Sera from patients as well as controls were tested for their level of immunologically reactive TNF-alpha using enzyme linked immunosorbant assay (ELISA). TNF-alpha levels in the sera of patients were significantly higher as compared to healthy control group. Acute infection was associated with the highest levels of TNF-alpha indicating that it may play a role in the pathogenesis of acute toxoplasmosis. In chronic infection, the level of TNF-alpha correlated with IHA antibody titre suggesting that, antibodies against T. gondii may participate in TNF-alpha production. It could be concluded that TNF-alpha may contribute to the pathology in acute infection and may play a role in modulating the host's immune defence against T. gondii in chronic infections. PMID- 8721229 TI - Serum IgG antibody response to Pneumocystis carinii among immunosuppressed, malnourished and healthy rats. AB - Pneumocystis carinii is an important opportunistic pulmonary pathogen that causing pneumonia in premature infants, children with immunodeficiency diseases and patients of all ages receiving immunosuppressive agents. In this work, humoral immune responses to this organism were studied, using IFAT during the period of infection and recovery of immunosuppressed and malnourished rats compared to healthy group. Where sever pneumonitis similar to that seen in humans can be induced in this experimental model. Serum IgG antibody titers to the organism were absent in rats administered corticosteroids but rose after steroid tapering with disappearance of the organism from their lungs due to reactivation of the immune system. While in malnourished infected rats, clearance of the organism had occurred after regaining the protein diet with progressive increase in IgG level denoting their immunocompetent state. IgG antibody appeared in the serum of control healthy rats with the progress of age at low non diagnostic level indicating the presence of dormant parasites in their lungs. PMID- 8721230 TI - On the efficacy of Co-trimoxazole as an oral treatment for pediculosis capitis infestation. AB - No doubt, pediculosis capitis or Pediculus h. capitis infestation is an increasing health problem particularly among Egyptian children. Many chemical pediculocides are used with a variety of effectiveness and side effects. Meanwhile, Co-trimoxazole (Trimethoprim and sulphamethoxazole) have shown some pediculocidal action. In this paper, Co-trimoxazole was given to patients infested with pediculosis capitis. Prolonged course was needed to free the patients from adult and nymphal stages but not the eggs (nits). The results were discussed. It was concluded that until the discovery of cheap, safe and effective oral drug, topical application of pediculocides as ointment or shampoo is the method of choice. PMID- 8721231 TI - Light and electron microscopic studies of Isospora chalchidis (Amoudi, 1989) infecting Chalcides ocellatus in Egypt. AB - Oocysts of Isospora chalchidis (Amoudi, 1989) were described from the faeces of the skink Chalcides ocellatus in Egypt. Non-sporulated oocysts were spheroidal, measuring 20 (18.5-21) microns in diameter and contained granulated zygotes. Sporulated oocysts had the same dimensions of the nonsporulated ones and each contained two sporocysts. Sporocysts were ovoid with stieda and sporocyst residual bodies. Sporulation time was 50 hours at room temperature. Merogony and gamogony occurred in the intestinal mucosa. Electron microscopic investigations showed that meronts, merozoites, gamonts and gametes developed in a narrow parasitophorous vacuole within the host-cell nucleus. Nuclei of meronts were surrounded by rough endoplasmic reticulum. Merozoites showed the main characteristics of motile stages of Apicomplexa. Macrogametes contained a large nucleus, two types of wall-forming bodies and a large amount of lipid inclusions. Nuclei of microgamonts were peripherally arranged and lacked nucleoli. Microgametes were flagellated. PMID- 8721232 TI - Host-parasite relationships of Wuchereria bancrofti and mosquito hosts, Culex pipiens L. and Aedes caspius pallas. AB - We compared the defense mechanisms directed against Wuchereria bancrofti by a filaria susceptible mosquito, Culex pipiens, and a refractory one, Aedes caspius. The reciprocal deleterious effects of both the mosquito host and its parasite are reported. Anatomical and histological examinations of mosquitoes revealed minor differences in the general structure of the foregut of either species. The cibarial pump of Ae. caspius, however, lacked a cibarial armature and damage inflicted to some of the microfilariae (mf) ingested by this mosquito was not mechanical. In contrast, the cibarial armature of Cx. pipiens consisted of 24 delicate teeth which did not seem to affect ingested mf. The peritrophic membrane (pm) did not constitute a significant barrier to mf migrating through the midgut wall of either mosquito. Indeed, mf forced their way out into the hemocoel, immediately after they have been ingested, and up to 60 min post-feeding, when the pm was still thin and soft. It hardened 16 h post-feeding. Traversing mf perforated the pm and displaced the basement membrane. Within the thorax of Cx. pipiens, the parasite completed its development synchronously. In contrast, mf ingested by Ae. caspius did not develop beyond the sausage stage, and many of these larvae were vacuolated. In both mosquitoes, parasitized thoracic muscle fibers were tunnelled and, only in Ae. caspius, their nuclei were pyknotic. No worm encapsulation or melanization was observed in either mosquito. It is concluded that defense lines to invading parasites in Cx. pipiens are insignificant, and that in Ae. caspius, the main barrier to parasite development resides within the thoracic muscles. In the absence of cellular evidence, it is suggested that such a barrier is physiological in nature. PMID- 8721233 TI - Cutaneous leishmaniasis in Egypt (review and comment). AB - Leishmania is primarily characterized by existing in two stages in its life cycle, each occurs in a distinct host. The amastigote stage found in the cytoplasm of the reticulo- endothelial cells, monocytes and other phagocytic cells of the vertebrate host. The promastigote stage found in the gut of its insect vector. The leishmaniasis comprise several diseases of wide diversity of manifestations caused by different species of the genus Leishmania. Because of the virtual morphological identity of the organisms throughout the genus, they are classified according to the clinical conditions which they produce in man, under three main headings: (1) Cutaneous leishmaniasis (CL.), (2) Mucocutaneous leishmaniasis (MCL.), (3) Visceral leishmaniasis (VL). Generally speaking, leishmaniasis is an example of a zoonosis that reaches man through an insect vector. The great majority of the Leishmania species are maintained by mammalian reservoir hosts in natural foci of infection. Rodents, dogs, wild cats, jackals, foxes, sloths, hyraxes and other carnivores are the animal reservoirs which maintain the infection in nature. The insect vectors are over 50 species of the genus Phlebotomus in the Old World and genus Lutzomyia in the New World. PMID- 8721234 TI - Prevalence of Schistosomiasis and dynamics of its distribution in some villages in Qalyoubia Governorate. AB - A total of 1535 persons of different age, sex and occupation from 5 villages (Tanan, El-Sad, Sandiun, Kalama and Taha-nob) in Qalyoubia Governorate in Nile Delta were subjected to clinical, stool, urine and intradermal tests accompanied by a questionnaire for 500 of them. Snail vectors of Schistosome parasites were collected from 2 villages (El-Sad and Taha-nob) and snail densities in the waterways as well as the prevalence rate of infection with schistosomal cercariae were recorded. The results revealed that the average prevalence rate of Schistosoma haematobium in the 5 cillages was 4.9% (range 2.3% - 7.1%) and that of S. mansoni was 13.3% (range 11.4-14.4%). The overall prevalence of both types was 18.2% (range 13.7-21.4%). The average density of Biomphalaria alexandrina in the 2 village was 16.47% and that of Bulinus truncatus was 7.5%, the ratio was 2:1. The infectivity with schistosomal cercariae was very low with an average of 2.5% in B. Alexandrina and 1.5% in B. truncatus. Factors responsible for the decreased rates of schistosomiasis and snail infectivity were discussed. PMID- 8721235 TI - Opportunistic intestinal protozoa in chronic diarrhoeic immunosuppressed patients. AB - Chronic diarrhoea accompanied by weight loss is a common and often debilitating problem in immunocompromized patients, receiving chemotherapeutic agents. In these patients, the intestinal opportunistic parasites probably played a major role in causing this clinical manifestation. The present work, aims to search for these parasites. Special stains for each parasite were used to differentiate it easily from the fecal elements, obviating the need for diagnostic invasive techniques especially used in microsporidial infection. The detected parasites were, Giardia lamblia (17.7%) best seen by iron haematoxylin stain. Coccidian oocysts (Cryptosporidia; 13.3% Isospora belli; 2.2%) were clearly seen by using Ziehl-Neelsen and Chromotrope-based stains. The Gram positive spores of Enterocytozoon bieneusi were (4.4%) and best seen by using chromotrope-based stain, where as Giemsa failed in their diagnosis. PMID- 8721236 TI - Leishmania major: amastigote formation in cell-free media. AB - The life cycle of Leishmania consists of two distinct developmental stages: the amastigote, which is the ovoid non-flagellated form found in the vertebrate host, and the promastigote, which is an elongated flagellated form found in the gut of an infected sandfly. Following its injection into the vertebrate host by the sandfly vector, the promastigote transforms into an amastigote after entering the host macrophage. The environmental cues inducing this transformation are not fully understood. Attempts to axenically develop and cultivate amastigotes from different Leishmania species have indicated that species and sometimes even strains of the same species vary in their requirements for this process (Pan et al., 1993). The majority of the available published data on transformation pertains to New World Leishmania and provides evidence that elevation in the incubation temperature and/or acidic pH can in some cases induce amastigote formation from promastigotes. PMID- 8721237 TI - Effect of ivermectin on survival and fecundity of Culex pipiens the vector of Wuchereria bancrofti in Egypt. AB - The effect of two different doses of ivermectin on the survival and fecundity of Culex pipiens was evaluated. Female mosquitoes (50 in each group), same age and generation, were fed once on blood from ivermectin-treated rabbits. Comparative treatments consisted of two different doses of ivermectin (0.1 mg or 0.4 mg/kg of body weight), and mosquito groups were fed on the rabbits at 3 days and 10 days post-treatment respectively. Mosquitoes were maintained with 10% sucrose solution, at 25 degrees C and 70% relative humidity. Observations were made at 7, 10, 15, 20, 25, and 30 days post-feeding. Mosquitoes fed on rabbits treated with the small dose at three days post-injection showed reduced survival of 94%, 62%, 44%, 8% and 0%, while the 0.4 while the 0.4 mg/kg of body weight (bw) dose reduced survival to 88%, 60%, 6% and 0%. Survival of the control groups were 100%, 90%, 88%, 68% and 52% respectively. Survival of adult females fed at the lower dose, 10 days post injection were reduced to 92%, 90%, 84% 60% and 44%; while the higher dose reduced survival to 86%, 58%, 34% 22% and 0%. Survival of control groups were 96%, 92%, 84%, 78% and 72% respectively. Fecundity resulting from blood meals at 3 and 10 days post injection were 61% and 85% for the lower dose, and 0% and 42% for the higher dose respectively. Hatching of eggs was 82% and 88% for the 0.1 mg/kg bw dose, and no hatching was observed at the higher dose. Larval survival rates resulting from blood meals taken 3 days and 10 days post-injection of the lower dose were 24% and 25% respectively. However, no larvae survived from the high dose groups. Larval survival rates in control groups were 42% and 40%, respectively. These results provide new information about the effect of ivermectin on the vector of lymphatic filariasis, and add a new dimension to the use of ivermectin in filariasis control. PMID- 8721238 TI - The immunological aspects of acute C.N.S. complications of the exanthematous viral diseases. AB - A total of 21 patients with postmeasles and 26 patients with postvaricella C.N.S. complications were studied. In both groups, males were predominant than females. The C.N.S. manifestations included disturbed level of consciousness, coma, seizures, motor deficits, ataxia and myoclonus. The sequelae were more frequent in postmeasles cases and ranged from behavioral abnormalities to motor deficits. C.S.F. examination showed that most of the cases demonstrated increase in the protein content (45-100mg) and pleocytosis. Myelin protein was detected in 8 samples and 6 samples of postmeasles and varicella C.S.F. out of 12 samples in Tested in each group. Specific virus IgG antibody was detected significantly in 8 paired C.S.F. samples of postvaricella group and only one sample of postmeasles out of 12 paired samples tested in each group. C-T. scan examination revealed that the most common finding was the brain oedema (13 in measles, and 21 in varicella group). PMID- 8721239 TI - Parasitic causes of hepatomegaly in children. AB - Three hundred children with hepatomegaly were selected. They were subjected to full clinical and laboratory examinations. Also serum samples were examined to detect IgG using ELISA against SEA, chromatography purified hydatid cyst antigen, commercially available Toxoplasma antigen, partially purified adult Fasciola antigen and second-stage larvae Toxocara canis antigen. IFAT was used to detect IgG against Toxoplasma and T. canis. A commercially available IHAT kit for leishmaniasis was used. Based on immunological assays, 125 cases were suffering from various parasitic infections. Thirty cases with schistosomiasis (10%), 26 cases fascioliasis (8.7%), 18 toxocariasis (6%), 35 toxoplasmosis (11.7%), 3 cases hydatidosis (1%) and 13 cases mixed parasitic infections. No parasitic causes could be found in 175 cases (58.3%). Moderate or marked hepatomegaly favours the presence of schistosomiasis. Whereas, most cases with other parasites and those with non-parasitic infections fall in the category of mild hepatic enlargement. There was no associated splenomegaly in cases with Fasciola, Toxocara, hydatid disease and/or the non-parasitic group. Most of hepatomegalic cases with non-parasitic causes were found to be associated with fever (88.5%). Fever was found in nearly 50% of cases with either Toxoplasma or Toxocara infections. Mild eosinophilia was found in all cases with parasitic causes. Only 24 cases of non-parasitic group (13.7%) had easinophilia. Moderate and high eosinophilia were found in cases with fascioliasis and toxocariasis. Cases with fascioliasis had a statistically significant increase in enzymes activities specially alkaline phosphatase. It was concluded that parasitic infections should be considered as an important cause of liver enlargement in children. Serological methods using purified antigenic fractions are an important tool for diagnosis. PMID- 8721240 TI - Biomphalaria glabrata: a new threat for schistosomiasis transmission in Egypt. AB - This is the first report on the infestation of irrigation and drainage systems at Giza, Qalyoubiya and Kafr El Shiekh Governorates, with Biomphalaria glabrata (the snail host of Schistosoma mansoni in the new world). Identification of this snail species was based on morphometry of the shell, shape of the radular lateral teeth and presence of the characteristic renal ridge. B. glabrata snails collected from natural water courses in Egypt proved to be susceptible to a local strain of S. mansoni. These snails were infected by 52% versus 75% for Biomphalaria alexandrina (the only local host) under the same laboratory conditions. Meanwhile the periodic total cercarial production was higher in B. glabrata than in B. alexandrina (252 +/- 110 cercariae/snail versus 203 +/- 86 cercariae/snail, respectively). Differences in infection rate, length of schistosome incubation period, duration of cercarial shedding and cercarial production in various sizes of the two snail species are reported and discussed. PMID- 8721241 TI - Studies on the helminth parasites of birds from Egypt: 1-trematoda and cestoda. AB - A total number of 1019 wild birds were examined, of which 486 (49%) were found infected with one or more species of helminth parasites. Digeneans were recovered from 151 (15%) and cestodes from 249 (24%) birds. Out of 20 helminth species recovered, 10 were found new (7 trematodes and 3 cestodes). The prevalence and the effect of concurrent infection on the most frequently occurring helminth parasites were recorded and discussed. A review of all the trematode and cestode parasites previously described from the Egyptian birds was also given. PMID- 8721242 TI - Detection of circulating Trichinella pseudospiralis antigen by enzyme linked immunosorbant assay (ELISA). AB - A modified double antibody sandwich ELISA was used as a sensitive technique for demonstrating the appearance of circulating antigen of T. pseudospiralis in the sera of infected mice. The sera were tested at 5 days, 2, 4, 6, 7, 8, 10 and 12 weeks P.I. The antigen was first detected on the 5th day P.I., its level increased to reach the 1st peak at 4 weeks, then declined at 6 weeks. A second peak was observed at 8th weeks followed by a sharp decline to give negative OD reading at 10th week and increased to a moderate level at 12 weeks P.I. PMID- 8721243 TI - Abnormal distribution of the histocompatibility antigens (HLA) in lousy patients. AB - The histocompatibility antigens have important functions in the development of the immune response, in the development of immunologic tolerance and in the resistance and susceptibility to diseases. In the present study, the frequency of the human leucocytic antigens (HLA) were studied in 31 lousy children with Pediculus h. capitis (head lice) and 14 adults with Phthirus pubis (pubic lice) to evaluate the immune response in their pathogenesis. The patients (children and adults) were parasite-free as indicated by urine, stool and blood analysis and clinical examination. A significant increase was found between HLA-A11 and, -B5 and lousy children with P. h. capitis and between HLA,-A11, -B5 and -B27 and lousy adults with P. pubis. The association between HLA antigens and parasitic infection was discussed. PMID- 8721244 TI - A comparative study on the efficacy of 0.4% Sumithrine and 1% Lindane in the treatment of Pediculus humanus capitis in Turkey. AB - Two different chemicals were used in the treatment of Pediculus h. capitis in cases, who were found to be infested during the screening of 1 secondary and 3 elementary schools in Bostanli-Karsiyaka, Izmir. Sumithrine was found to be more effective than Lindane according to the follow up control performed a month after the treatment. While the efficacy of Sumithrine in Emlakbank elementary school, Metin Asikoglu elementary school and Emlakbank secondary school was noted as 93%, 96% and 86.8% respectively. The efficacy of Lindane was found to be 67.5% in Murside Akyuz elementary school. PMID- 8721245 TI - Comparison between the prospect Cryptosporidium Microtiter Assay and the immunofluorescent test in the diagnosis of cryptosporidiosis. AB - The immunofluorescent test utilizing rabbit antisera against Cryptosporidium oocysts isolated from dairy calves was compared with the proSpect Cryptosporidium Microtiter Commercial Assay which is an enzyme immunoassay utilizing monospecific antibody to Cryptosporidium specific antigen (CSA). The former test proved to be by far more sensitive than the latter one in diagnosing human cases positive for cryptosporidiosis by the modified acid fast stain. PMID- 8721246 TI - Filtration, centrifugation and mouse exposure for the detection of schistosome cercariae in water. AB - Three methods for detection of schistosome cercariae in water were studied under laboratory and simulated field conditions in Egypt. Water samples tested by filtration and centrifugation quantitatively reflected the density of cercariae in moderately turbid canal water (30 Nephelometry Turbidity Unites, NTU). When tested in waters of increasing turbidity, the centrifuge gave more consistent results than the filtration system. The centrifugation technique was more efficient in processing the water samples and took almost half the time of filtration. Mouse exposure was useful in detecting cercariae, but was not quantitative. However, there was a significant improvement in worm recovery when mice were more close together in the floatation device. Mouse exposure was also expensive and cumbersome to use. It appears from the present results that the technique of choice for further cercariometric work under Egyptian conditions may be centrifugation. PMID- 8721247 TI - Effect of mebendazole and ivermectin in experimental hepatic capillariasis: parasitological, scanning electron microscopy and immunological studies. AB - In this study, mebendazole and ivermectin were tried during three different phases of C. hepatica infection. At an early phase, when immature forms were present both drugs were effective in causing destruction and degeneration of the larvae, thus preventing subsequent growth and maturation and consequently the complete absence of eggs. During the second phase, which is found to be the most critical period the two drugs used led to degeneration and resorption of most of adult worms. In the third phase both mebendazole and ivermectin were effective in decreasing the mean number of eggs. After treatment the topographic changes were in the form of disorganized cuticle of the worms and absence of surface uniformity. Such a disorganized cuticle is vulnerable to be attacked. C. hepatica eggs showed irregularities and longitudinal grooves indicated shrinkage of the shell. The effect of the two drugs indicate that both of them were effective in the treatment of hepatic capillariasis. PMID- 8721249 TI - Effect of the behavioural stress on susceptibility of Syrian hamsters to Schistosoma mansoni infection: effect on number and fertility of worm burden. AB - Effect of pairing on susceptibility of male Syrian hamsters to Schistosoma mansoni infection in comparison to their isolated counter-parts have been studied. Pairing resulted in significant differences between paired and isolated hamsters in measurements of eggs, miracidia, number of adult worms recovered and mean testicular number of males. Paired hamsters seemed to be immunologically suppressed and such a suppression resulted in increasing the fertility and number of worms recovered. The width of the male schistosomes recovered from the paired hamsters was two folds and the length was four folds greater than those recovered from isolated ones. The mean testicular number was 8.17 +/- 0.53 in males recovered from paired hamsters and 6.16 +/- 0.71 in those recovered from isolated ones. Of interest was, the demonstration of a male with two groups of testes recovered from a paired hamster. It was concluded that the stressed host reflex effects on the number and fertility of the schistosome worms burden, and hence the severity of the disease and its complications. PMID- 8721248 TI - Detection of Pneumocystic carinii in immuno-suppressed rats by different histological stains and immunological assays. AB - Detection of P. carinii in lung sections of 35 immuno-suppressed albino rats was evaluated using five histological stains (toluidine blue "O", Giemsa, Gram's stain, PAS and H & E) and two immunological assays (indirect immunofluorescence [IIF],, and indirect immunoperoxidase [IIP]) using polyclonal antibody, raised in white New Zealand rabbits, against rat P. carinii. In spite of the high sensitivity of the histological stains, they yield hazy details insufficient for easy identification with difficult interpretation. However, Gram and PAS stains provided a better visualization with easy identification of the parasite. On the other hand, both immunological assays allowed an accurate rapid interpretation of the stained slides. The IIP technique, having 100% sensitivity in detecting the precipitated parasite antigens with absolute specificity, seems to be a good diagnostic tool for detecting P. carinii in lung sections. PMID- 8721250 TI - Indapamide treatment modifies the vascular reactivity of normotensive rat mesenteric arteries ex vivo. AB - Indapamide is an antihypertensive diuretic drug of which the precise antihypertensive mechanism has not been clarified. The aim of the present study was to investigate with mesenteric arteries of normotensive rats the possible change in vascular reactivity ex vivo and in vitro after indapamide application, taking into special consideration the role of endothelium. After seven days of drug treatment in vivo with low-dose 0.3 mg/kg/day or high-dose 3 mg/kg/day of indapamide the effect on the vascular responses ex vivo were compared to controls. The contractions induced by noradrenaline (NA) or potassium chloride (KCl) were increased in vascular rings without endothelium in the high-dose indapamide group. No alterations in endothelium-dependent relaxation responses to acetylcholine (ACh) or endothelium-independent responses to sodium nitroprusside (SNP) were found between any of the treatment groups in vascular rings with or without endothelium. When indapamide (1 microM or 10 microM) was added in vitro for 10 min preincubation it did not significantly change the vascular contractions induced by cumulatively increased concentrations of NA or KCl. The relaxation responses induced by ACh or SNP were also not affected by indapamide pretreatment. Our results suggest that the lack of change in contractile response in vitro after indapamide pretreatment might be related to the prodrug nature of indapamide, which is not metabolized during the short preincubation. PMID- 8721251 TI - Enhanced ornithine decarboxylase activity of chick muscle cells in culture by beta-adrenergic agonists. AB - The ability of beta-adrenergic agonists to stimulate ornithine decarboxylase activity (ODC) in chick muscle cell culture prepared from 11-day old embryos was evaluated. After 72 h of preincubation (myotube formation) the medium was supplemented for 4 h with noradrenaline, ritodrine, isoproterenol or clenbuterol, at concentrations of 10(-12), 10(-9) and 10(-6) mol/l. No significant response of ODC activity to noradrenaline was observed. The highest concentration (10(-6) mol/l) of the beta-adrenergic agonists ritodrine and isoproterenol elevated the activity of ODC. Clenbuterol was the most active beta-adrenergic agonist. The lowest concentration (10(-12) mol/l) had an apparent effect on ODC activity in muscle cell culture, and the substitution of media at levels of 10(-9) and 10(-6) mol/l had a similar effect in comparison to controls. The potency of beta adrenergic agonists in increasing ODC activity was on the following order: noradrenaline, ritodrine, isoproterenol, clenbuterol. Results indicate that beta adrenergic agonists may directly stimulate ODC activity followed by physiological processes in the muscle cells in the early stage of chick embryonic development. PMID- 8721252 TI - Effects of betahistine, a histamine H1 agonist and H3 antagonist, in a light/dark test in mice. AB - The effects of betahistine, a histamine H1 agonist and H3 antagonist, were investigated in a light/dark test measuring anxiety in mice. Betahistine significantly decreased the locomotion and rearing in the light and dark zones, shuttle crossing and time spent in the light zone in the light/dark test. These phenomena suggest that betahistine has anxiogenic effects and/or locomotor suppressive effects. Pyrilamine (6 mg/kg) and ketotifen (6 mg/kg), H1-receptor antagonists, antagonized the effects of betahistine (100 mg/kg) on all parameters in the light/dark test but zolantidine (6 mg/kg), an H2-receptor antagonist, augmented its effects on some parameters decreased by betahistine. Especially, it potentiated the decreases by betahistine in the parameters showing anxiety in this test. The decreases in the light/dark test induced by zolantidine (6 mg/kg) plus betahistine (200 mg/kg) were antagonized by pyrilamine (12 mg/kg) but antagonism of the decrease in the time spent in the light zone as a parameter showing anxiety was not significant. These results suggest that the effects via H1 receptors are involved in anxiety in the light/dark test in mice but the other factor may be simultaneously needed for induction of behaviorally detectable anxiogenic effects in this test. PMID- 8721253 TI - Role of inward and delayed rectifier currents in generation of early afterdepolarization in guinea pig ventricular myocytes under K(+)-free or low K+ superfusion. AB - Early afterdepolarization (EAD) was studied in isolated ventricular myocytes of guinea pig heart. Under K(+)-free's treatment, most of the myocytes showed hyperpolarization in resting potential and the duration of action potential was prolonged, eventually leading to the appearance of EAD with the second plateau of -76 +/- 3 mV. TTX (10 microM) and verapamil (10 microM) or normal Tyrode's solution abolished the EAD. The background I-V curve showed inward rectifying with a crossover in the level of -80 to -30 mV and the reversal potential shifted from -80 to -120 mV when normal Tyrode's solution was changed to K(+)-free solution. The changes of I-V relationship of inward current IK) were similar to the background ones except without crossover. The delayed rectifier current (IK) was inhibited significantly under K(+)-free treatment. Low K+ (2.7 mM) superfusion was able to induce EAD in only a few cases (4/15). Adding Cs+ (5.0 mM) into low K+ solution, EAD was induced in almost every case. The background I V curve was inhibited slightly under low K+ superfusion, but was inhibited significantly with a remarkable crossover under low K+ and Cs+ treatment. The changes of I-V curve IK1 under low K+ or low K+ and Cs+ treatment were similar to the changes of background ones. There were no significant changes in the IK under low K+ superfusion while a remarkable inhibition occurred under low K+ and Cs+ treatment. It was suggested that both IK1 and IK were involved in the induction of EAD under K(+)-free or Cs+ treatment in guinea pig ventricular myocytes. PMID- 8721254 TI - Interactions between histaminergic and cholinergic pathways of gastric motility regulation. AB - Smooth muscle preparations, isolated in a circular direction from guinea pig gastric fundus, were used to study the effects of H1 and H2 antagonists on acetylcholine (ACH)- and histamine (HA)-induced contractions as well as the effects of HA antagonists on spontaneous contractile activity. HA (1 x 10(-9) M to 1 x 10(-5) M) concentration-dependently enhanced the tone of the strips with ED50 = 3.5 x 10(-7) M. Applied 5 min before HA, the H1 antagonists (mepyramine, diphenhydramine, dimethpyrindene) and the H2 blockers (ranitidine, cimetidine, roxatidine) reduced HA-induced contractions. HA in concentrations of 1 x 10(-8) M to 1 x 10(-7) potentiated, and in higher concentrations (1 x 10(-6) M to 1 x 10( 5) M) inhibited, smooth muscle contractions evoked by low frequency electrical field stimulation (EFS). The H1 blockers (1 x 10(-6) M to 1 x 10(-4) M) concentration-dependently enhanced smooth muscle tone, the maximum contractions being about 50% smaller than the contractile responses to 1 x 10(-5) M ACH and 5 x 10(-5) M HA. Tetrodotoxin, atropine and indomethacin shifted to the right the concentration-response curve for mepyramine, reducing its maximum by 25, 58 and 62%, respectively. The H2 blocker ranitidine also suppressed (by 42%) mepyramine evoked increase in the fundic strips tone. The H1 antagonists reduced ACH-induced contractions of the smooth muscle strips and did not affect the contractions in response to EFS. The H2 blockers had no effect on tone and ACH-evoked contractions of the smooth muscle strips but concentration-dependently enhanced both the contractions and [3H]-ACH release in response to EFS. The results demonstrate the presence of both H1 and H2 postsynaptic receptors which are involved in the direct myogenic action of HA on guinea pig gastric fundus smooth muscles. It also appears that HA might concentration-dependently modulate the cholinergic neurotransmission in gastric fundus. It could be suggested that H1 blockers have a direct myogenic effect on guinea pig gastric fundus smooth muscle and might also interact postsynaptically with muscarinic receptors in this tissue. PMID- 8721255 TI - Pharmacological characteristics of Sho-seiryu-to, an antiallergic Kampo medicine without effects on histamine H1 receptors and muscarinic cholinergic system in the brain. AB - The pharmacological characteristics of Sho-seiryu-to, an antiallergic Kampo medicine, were investigated. Forty-eight-hour passive cutaneous anaphylactic (PCA) reaction was significantly inhibited in rats orally administered Sho-seiryu to (1000 mg/kg). Sho-seiryu-to significantly inhibited increase in vascular permeability induced by histamine. These data confirm previous findings that Sho seiryu-to has antiallergic activity in animals and suggest that the antagonism of histamine may be an antiallergic mechanism of Sho-seiryu-to. Sho-seiryu-to did not affect locomotor activity or motor coordination in mice. Although ketotifen prolonged sleeping time induced by pentobarbital, Sho-seiryu-to had no such effect. Nor was there any effect on oxotremorine-induced tremor and [3H] mepyramine binding to histamine H1 receptors in rat brain. Thus, Sho-seiryu-to appears to be useful for treating type I allergy, with relatively few side effects such as sedation and drowsiness due mainly to blockade of histamine H1 and muscarinic receptors in the brain. PMID- 8721256 TI - Urinary excretion of meperidine and normeperidine in man upon acute and chronic exposure to high altitude. AB - The urinary excretion of unchanged meperidine (M) varies with change of pH and metabolism. Since exposure of man to high altitude (H) may cause significant physiologic changes, we investigated its effects on the urinary excretion of M. The study was carried out in 3 groups of healthy, male volunteers (ages 18-20 years): at sea level (L), at 4360 m the day after arrival at H (HA), and at 4360 m in subjects residing for > 10 months at H (HC). Urine was collected for the periods of 0-4, 4-8, 8-12 and 12-24 h. Urinary pH was measured and the concentrations of M and normeperidine (N) were determined. The 24 h excretion of M and N was significantly decreased for L vs. HA and L vs. HC. Significance was also seen for the periods 0-4, 4-8 and 8-12 h. The ratio of amount excreted M/N for the 24 h period was highly significant for L vs. HA and L vs. HC. The urinary pH ranged from 5.3-5.9 for L, 5.9-7.0 for HA, and 5.1-5.7 for HC. The Fel (fraction of M eliminated in the unchanged form in urine) significantly decreased from L to HA and HC. PMID- 8721257 TI - Effects of the novel neuroprotective agent, riluzole, on human brain function and behavior: I. Double-blind, placebo-controlled EEG mapping and psychometric studies under normoxia. AB - In a double-blind, placebo-controlled, crossover study, the encephalotropic and psychotropic properties of single oral doses of the novel neuroprotective agent, riluzole, were investigated utilizing EEG mapping and psychometry. Twenty healthy young volunteers received randomly at weekly intervals, placebo, 50, 100 and 200 mg riluzole. EEG recordings and evaluation of 9 noopsychic and 5 thymopsychic variables were carried out at 0, 2, 4, 6 and 8 h after oral drug administration. EEG maps on the multivariate analysis demonstrated that all three doses induced significant changes in human brain function, as compared with placebo, between 2 and 8 h, with effect only increasing slightly with dose. EEG maps on univariate analysis demonstrated generally an increase of delta/theta, decrease of alpha and beta power, as well as a slowing of the centroid of the total power spectrum, which suggests sedative properties of the drug. Only after the two highest doses at 6 h were some different findings observed. Multivariate statistics on psychometry failed to show any significant effects on the noopsyche, while for the thymopsyche, all three doses of riluzole produced a deterioration. The latter was characterized by a decrease in drive and wakefulness as well as deterioration in well-being, mood and affectivity. Thus, under normoxia, in all three doses riluzole produced neurophysiologically a sedative effect, accompanied at the behavioral level by a deterioration in the thymopsyche, which may be expected from a drug with antiglutamatergic effects in normals. PMID- 8721258 TI - Effects of the novel neuroprotective agent, riluzole, on human brain function and behavior: II. Double-blind, placebo-controlled EEG mapping and psychometric studies under hypoxia. AB - In a double-blind, placebo-controlled study, the antihypoxidotic properties of the novel neuroprotective agent, riluzole, were investigated utilizing blood gas analysis, EEG mapping and psychometry under a transient, reversible, hypoxic hypoxidosis. The latter was induced by a fixed gas combination of 9.8% oxygen (O2) and 90.2% nitrogen (N2) (found at 6000 m altitude), which was inhaled for 23 min under normobaric conditions by 20 healthy, young volunteers. They randomly received, after an adaptation session, single oral doses of placebo, and 50, 100 and 200 mg riluzole. Evaluation of blood gases, EEG mapping and psychometry were carried out 0, 2, 4, 6 and 8 h postdrug, each time under the 23-min hypoxia. Blood gas analysis demonstrated a drop in PO2 from 106 to 37 and 36 mmHg, in PCO2 from 35 to 31 and 31 mmHg at 14 and 23 min of inhalation, respectively, while pH increased from 7.43 to 7.48 and 7.48. Base excess and standard bicarbonate remained stable. EEG mapping exhibited under hypoxia a marked increase of delta/theta, decrease of alpha and an increase of superimposed beta activity, as well as a slowing of the centroid of the total activity, which reflects deterioration of vigilance. Riluzole in lower doses and at early hours after higher doses did not attenuate this hypoxia-induced vigilance decrement, while with higher doses (100-200 mg) in later recording periods (6-8 h) brain protection occurred. As compared with placebo, delta/theta power increased at 2-8 h after 50 mg riluzole and up to 4 h after 100 mg riluzole, while a decrease occurred at 4 and 8 h after 100 mg and at 6-8 h after 200 mg. Alpha power showed no changes after 50 mg, an increase at 2 and 8 h after 100 mg and a decrease at 4 h after 200 mg, with no changes thereafter. Beta power decreased at various times after all three doses. At the behavioral level, hypoxic hypoxidosis induced a deterioration of the noopsyche, which was not mitigated by riluzole. In regard to the thymopsyche, there was even a slight deterioration after all three doses, as compared with placebo. PMID- 8721259 TI - [In our own business: economic analyses in radio-oncology]. PMID- 8721260 TI - [The importance of a quality standard and performance numbers in radio-oncology]. PMID- 8721261 TI - [Quality and the profit situation in ambulatory radiotherapy]. AB - BACKGROUND: Increasing demands of quality assurance and growing complexity of the concepts of radiotherapy planning produce increasing costs. These costs, on the other hand, cannot be got back from the paying institutions, because, in general, treatment sessions are paid in a lump sum. Using examples from everyday's practice, it will be shown, in which range levels of quality have an influence on profit. METHOD: For 4 frequent indications for curative radiotherapy: NSCLC, radiotherapy after breast-conserving operation for breast cancer, cancer of the hypopharynx and cancer of the uterine cervix, treatment plans on 3 different quality levels have been created. For each treatment step, times in minutes have been measured and costs calculated according to staff, machines, and rooms. RESULT: Rising quality levels implies, in general, lowering the profit. CONCLUSIONS: Radiotherapists have to define in their workshops, which quality level is tolerable, essential, desirable or an option for the future. The single radiotherapist, then, has to decide, on what quality level he is able to work, and which radiotherapy techniques he cannot afford to offer in general or at certain times. The professional organisations have the difficult task to fight for a level of reimbursement allowing the further evolution of our profession. PMID- 8721262 TI - [Costs-proceeds calculation for radiotherapy; comment]. AB - BACKGROUND: The Health Care Structure Bill (Gesundheitsstrukturgesetz) and the Federal Health Care Tariff Regulation (Bundespflegesatzverordnung) pose new challenges for the chief physician in charge, which he has to face on account of hospital management control with department-specific internal budgeting of services, expenditure and remittance. In view of the impending establishment of care-related lump sums (Fallpauschalen) and special allowances for radiation therapy (Sonderentgelte) as well as the introduction of an appropriate internal budget plan, we resolved to draw up the service-cost ratio for 11 tumor entities, implementing calculation based on process evaluation, and to match this calculation to the actual proceeds from outpatient treatment covered either by general health insurance or private care insurance. MATERIAL AND METHODS: The following 11 tumors were included in the analysis: Glioblastoma multiforme, squamous cell carcinoma of the head and neck, lung cancer, oesophageal cancer, breast cancer, rectal and prostate cancer, cancer of the anal canal and the uterine cervix, Hodgkin's disease and bone metastases. For the description of the services rendered and for the calculation of actual cost, the complex treatment regime was divided into the following steps: administration, medical radiation planning, radiation planning by physicist, workmanship, radiotherapy treatment and medical service. The calculation of costs was made either in- or excluding the cost of teatment facilities and basic investment. The proceeds were calculated according to the regulations of the EBM (dated 1. 10. 1994) for general health insurance patients (median point value 9.35 DPF) and the GOA (dated 1. 7. 1988) for private patients (point value 11 DPF). RESULTS: 1. Calculation by EBM standards: Excluding the cost of facilities and basic investment, there is a profit margin for the following tumor entities amounting to +1.6% (103.18 DM) for ENT tumors, +9.2% (671.90 DM) for carcinoma of the anal canal and +35.8% (1637.84 DM) for bone metastases. All other tumor entities cannot be treated cost-effectively. With the exception of glioblastoma, median loss comes to -20% (-14% to -27.4%, i.e. -809.48 to -1812.84 DM). Including in the calculation the cost of the facilities and investment, only bone metastases can be treated with a profit margin. For all other tumors, the mean loss amounts to -33% (-14.1% to -44.7%) or else -1308.26 to -4550.51 DM. 2. Calculation by GOA standards: Comparing cost and actual proceeds, there is a loss for all tumor entities. Excluding the cost of treatment facilities and investment, median loss amounts to -44% (-33.3% to -53.5%, or -1752.64 DM to -3488.18 DM), including both, mean loss comes to -56% (-47.8% to -53.3% or -3156.33 DM to -6332.43 DM). 3. Calculation of comprehensive cost and of the proceeds: For our own department, considering the cost-proceeds ratio for 1994, we ran up a deficit of 673,767.10 DM for outpatient care. This deficit can mainly be traced back to non-operational hours of the linear accelerator due to machine impairment, maintenance and dosimetric measurements by the physicists, leading to the loss of 53 work-days (14 hours each) amounting to 498,771.34 DM plus general costs of 271,000 DM added to the bill by hospital administration. CONCLUSIONS: As to the reviewed tumor entities, modern standard radiotherapy cannot be administrated cost-effectively. A considerable degree of under-funding is especially evident with the inclusion of cost of investment and of the facilities. The cost-proceeds ratio is most unsatisfactory as regards GOA calculation, which is obsolete as far as service description and service payment is concerned. As it in no way measures up to the required standard of modern radiotherapy, reform is therefore urgently needed. PMID- 8721263 TI - [The personnel needs of health physics in radiotherapy]. AB - METHODS AND RESULTS: Using a questionnaire, mean occupation time values for the different medical physics activities were derived in 1992; they formed the basis for recommendations of minimum physics staffing levels in radiotherapy. The recommended staffing levels were compared with the actual staffing levels and to other national and international recommendations. PMID- 8721264 TI - Influence of ouabain on cell inactivation by irradiation. AB - BACKGROUND: It has been suggested that irradiation affects the function of the Na(+)-K(+)-ATPase. Here we examine the influence of the inhibitor ouabain on the cytotoxicity of irradiation. MATERIAL AND METHODS: Cell colony assay, cell survival, 86Rb-uptake, flow cytometry. RESULTS: In V79, HeLa and A549 cells ouabain alone causes a significant growth reduction at medium concentrations of 10(-4) M, 10(-6) M and 10(-7) M, respectively. When cells were exposed to the drug for 1 h and subsequently irradiated, the SF2 values decreased from 0.55 to 0.41, from 0.42 to 0.18 and from 0.57 to 0.35 in V79, HeLa and A549 cells, respectively. These effects were manifest at drug concentrations of 10(-3) M, 10( 6) M and 10(-7) M respectively, where Na(+)-K(+)-ATPase activity as measured by 86Rb-uptake was reduced to 40 to 60% of the control value. Addition of the drug after irradiation and when the G2/M cell cycle block was firmly established, markedly delayed the recovery of cells for well over 6 h and G1 levels remained at 50% of the control values. CONCLUSION: It is concluded that ouabain is strongly dose modifying in the human cell lines HeLa and A549 at concentrations which correlate with the inhibition of the Na(+)-K(+)-ATPase. Ouabain also inhibits the recovery of cells blocked in the cell cycle by irradiation. PMID- 8721265 TI - [Supportive treatment with megestrol acetate during radio(chemo)therapy in patients with tumors in the head-neck area. A randomized study]. AB - BACKGROUND: The value of megestrol acetate in treating tumor anorexia and cachexia of terminal patients is well known. However, the supportive effect of megestrol acetate during intensive radio-(chemo-)therapy was not investigated up to now. Therefore a randomized trial was performed including patients with advanced tumors in the head and neck region. PATIENTS AND METHODS: From June 1991 to December 1993 a total of 64 patients were admitted to a randomized, double blind placebo-controlled study. During and up to 6 weeks following radiotherapy patients received 160 mg/d megestrol acetate or placebo. The nutritional status (anthropometric and laboratory parameters) and the quality-of-life index according to Padilla et al. [24] were determined prior to therapy, 1, 4, 6 weeks later during radiotherapy and 12, 18 weeks after completion. RESULTS: Sixty-one out of 64 patients were evaluable (control group: n = 30; megestrol acetate patients: n = 31). One patient refused further participation after randomization. One patient in each arm was excluded due to side effects (impotence, diarrhoea). Further side effects were not observed. In the control group the nutritional parameters (body weight, triceps skinfold) and the subjective feeling of the patients deteriorated during radiotherapy and did not restore following radiotherapy. By contrast, the patients of the megestrol acetate group were able to stabilize these parameters. This difference was most prominent in the orally nourished patients (weight loss during therapy: control group: -4.1 kg; megestrol acetate group: -0.8 kg; p = 0.004); but not in the patients fed by percutaneous endoscopically guided gastrostomy (weight loss control group: -2.4 kg; megestrol acetate group: -0.8 kg; p = 0.14). CONCLUSION: In patients on radiochemotherapy megestrol acetate prevents patients from further deterioration of the nutritional status and quality of life. PMID- 8721267 TI - [Systemic neoadjuvant chemotherapy in advanced head and neck tumors: a randomized study of the combination of 5-FU/carboplatin versus 5-FU/cisplatin]. PMID- 8721266 TI - Radiation therapy for early glottic carcinoma (T1N0M0). The final results of prospective randomized study concerning radiation field. AB - PURPOSE: To investigate the effect of radiation field on the local control of early glottic carcinoma by prospective randomized study. PATIENTS AND METHODS: From May 1982 through February 1992, a total of 273 patients with early glottic carcinoma (T1N0M0) was treated at our department with a wedge filter technique using a shell to improve the dose distribution and immobilization during radiotherapy with 4 MV X-ray. Patients were randomly allocated to either treatment group A (radiation field size: 5 x 5 cm) or B (6 x 6 cm) using bilateral parallel opposed portals. Total radiation dose administered was 60 Gy in 30 fractions over a 6-week period. RESULTS: The 5-year recurrence-free survival rates were 88% in groups A and B (no significant difference). Minor chronic complication such as persistent arytenoid edema lasting more than 6 months or benign polypoid lesion of vocal cord was more frequently observed in group B (23%) than in group A (17%) (p = 0.038) while acute mucosal reaction and skin reaction showed no significant differences. CONCLUSION: A small field (5 x 5 cm) with an appropriate angle of wedge filter and shell fixing device is recommended to avoid adverse effect with keeping local control of early glottic carcinoma. PMID- 8721268 TI - [The different depths of inguinal lymph nodes and their importance in the planning of the irradiation of vulvar carcinoma]. PMID- 8721269 TI - [Does prophylactic whole-brain irradiation in small-cell bronchial carcinoma at the "limited disease" stage improve overall survival?]. PMID- 8721270 TI - [Acute gastrointestinal side effects in radio-oncology--what is certain in therapy?]. PMID- 8721272 TI - Prospective evaluation of a dose prediction algorithm for intravenous tobramycin in adolescent and adult patients with cystic fibrosis. AB - The predictive performance of a new algorithm to calculate the initial daily dose of tobramycin in patients with cystic fibrosis (CF) was prospectively evaluated. Twenty-six patients with CF (15 men, 11 women, 18-45 years of age) with an acute exacerbation of their chronic pulmonary infection were treated with intravenous tobramycin. The initial dose was calculated with a previously presented algorithm. This algorithm was derived from correlation analysis performed on the adjusted daily dose guided by the determination of serum concentrations: dose (mg three times daily) = 90 + 2.13 x LBM (kg), where LBM (male) = (1.1 x body weight [BW]) - (128 x BW2/height2) and LBM (female) = (1.07 x BW) - (148 x BW2/height2). The predictive performance of this algorithm was evaluated comparing the calculated initial daily dose with the adjusted daily dose for peak and trough levels of 9-11 mg/L and 1.0 mg/L, respectively. Mean squared error and mean error were determined as reflections of precision and bias. The predictive performance of the algorithm was compared with historical data on the predictive performance of the standard equation to dose of 3.3 mg/kg body weight three times daily. The dose calculated with the algorithm proved to give peak serum concentrations in a narrower range and to have a greater precision, but bias was equal. Applying the algorithm, more patients had initial peak serum concentrations in the pre determined range of 9-11 mg/L than when using the standard equation, so fewer dose adjustments had to be made. PMID- 8721271 TI - The effect of age and concomitant treatment with other psychoactive drugs on serum concentrations of citalopram measured with a nonenantioselective method. AB - We measured citalopram and desmethylcitalopram concentrations in serum from 169 psychiatric patients, who were treated with common therapeutic drug doses. Altogether 202 serum samples were assayed by a nonenantioselective high performance liquid chromatography (HPLC) method. The results indicate that the kinetic variability (maximum concentration/minimum concentration) in dose- and weight-related serum citalopram (10.6-fold) and desmethylcitalopram (7.2-fold) is large even during monotherapy. Log serum citalopram (r = 0.36, p < 0.05) and desmethylcitalopram (r = 0.51, p < 0.01) concentrations of individual patients increased significantly with increasing drug doses. Dose- and weight-related (calculated as mg/kg dose basis) log serum citalopram (r = 0.29) but not desmethylcitalopram (r = 0.06) concentrations increased with aging (p < 0.001). No sex-related differences were found. Nineteen patients (19 samples) had concomitant treatment with neuroleptics, 84 patients (101 samples) with benzodiazepines, and 18 patients (28 samples) with tricyclic antidepressants. The concentrations in these patients were compared with those of 48 nonsmoking patients (54 samples) without any concomitant psychotropic drug treatment. None of the single neuroleptics alone had a significant effect on dose- and weight related serum citalopram or desmethylcitalopram concentrations. However, citalopram concentrations increased by 121% (338 +/- 165 vs. 747 +/- 505, mean +/ SD; p < 0.01) and desmethylcitalopram by 85% (124 +/- 53 vs. 229 +/- 138; p < 0.05) when neuroleptics were pooled. Among single benzodiazepines, only alprazolam increased serum citalopram (338 +/- 165 vs. 391 +/- 267; p < 0.01) and desmethylcitalopram (124 +/- 53 vs. 186 +/- 175; p < 0.01) concentrations. When all the benzodiazepines were pooled, they still increased the serum concentration of the parent drug by 23% (338 +/- 165 vs. 414 +/- 303; p < 0.05) and those of the metabolite by 47% (124 +/- 53 vs. 182 +/- 163; p < 0.01). In patients who were simultaneously treated with clomipramine, serum citalopram (338 +/- 165 vs. 655 +/- 409; p < 0.001) and desmethylcitalopram (124 +/- 53 vs. 435 +/- 347; p < 0.001) concentrations were consistently higher than those of the controls. Even when the tricyclic antidepressants were pooled, they increased citalopram concentrations by 44% (338 +/- 165 vs. 486 +/- 312; p < 0.001) and desmethylcitalopram concentrations by 111% (124 +/- 53 vs. 261 +/- 260; p < 0.001). The results suggest that interindividual variability in serum citalopram concentrations is pronounced and that increased serum citalopram levels are related to advancing age and concomitant treatment with other psychotropic drugs. The citalopram dose should therefore ideally be individualized by therapeutic drug monitoring. PMID- 8721273 TI - Lack of effect of antimycotic itraconazole on the pharmacokinetics or pharmacodynamics of temazepam. AB - The azole antimycotics itraconazole and ketoconazole are potent and relatively nonspecific inhibitors of cytochrome P450 enzymes and have a potentially dangerous interaction with midazolam and triazolam. The possible interaction between itraconazole and a short-acting benzodiazepine, temazepam, was investigated in a double-blind, randomized crossover study. Ten healthy volunteers were given placebo or 200 mg itraconazole a day orally for 4 days. The challenge dose of 20 mg of temazepam was ingested on the fourth day, after which plasma samples were collected, and psychomotor performance tests were carried out for 24 h. Despite a statistically significant small increase of the area under the temazepam concentration-time curve, there was no clinically significant interaction, as determined by the psychomotor performance tests. The different metabolic pathways and the lack of significant firstpass metabolism of temazepam explain the great difference in the interaction potential of temazepam compared with midazolam and triazolam. Temazepam, unlike midazolam and triazolam, can be prescribed in usual doses for patients receiving itraconazole and other inhibitors of P450 3A4. PMID- 8721274 TI - Total and free methotrexate pharmacokinetics in rheumatoid arthritis patients. AB - The pharmacokinetics of total and free methotrexate (MTX) were investigated in 50 patients with rheumatoid arthritis. Each patient received 10 mg MTX intramuscularly. The free and total plasma concentrations of MTX were measured over a 36-h period after drug administration by using the Abbott TDx fluorescence polarization immunoassay. Plasma concentrations of MTX were described by a biexponential function. The mean terminal elimination half-lives of total and free MTX were 9.4 and 8.4 h, respectively, and the corresponding mean residence times, 8.5 and 9.2 h. No difference in these parameters was found by comparing total and free MTX. Total plasma clearance of the free fraction averaged 215 ml/min. The statistical comparison of the variations with time of the ratio of free to total MTX during the 36 h after the dose showed that the free fraction was significantly increased for 8 h after drug administration (p < 0.001). To describe these variations, the changes of the free MTX concentrations (unbound) were related to the changes of the total MTX concentrations by using the Hill equation. Mean plasma protein binding ranged from 20 to 57% for these patients. PMID- 8721276 TI - Influence of pharmacokinetic model on vancomycin peak concentration targets. AB - The aim of this study was to adapt the vancomycin therapeutic range to the kinetic models usually employed in clinical settings (one- and two-compartment models). Estimates of vancomycin pharmacokinetic parameters were obtained for both models in 22 hematologically malignant patients on vancomycin treatment using two serum concentrations and a bayesian algorithm. From these individually estimated pharmacokinetic parameters, an estimation of the maximum (Cssmax), 2 h postinfusion (Css2), and minimum (Cssmin) steady-state vancomycin serum concentrations for the one- and two-compartment models was made for a fixed 30 mg/kg/day dose. The linear regression equations between the predicted Css2 and Cssmin for the one- and two-compartment models do not differ significantly from the identity line, whereas the corresponding equation for Cssmax points to a 61% underestimation of Cssmax when the one-compartment model is used. From this latter regression equation, it is possible to define 20 mg/L (range of 18-21 mg/L) as a target Cssmax vancomycin serum concentration when a one-compartment model is used to monitor vancomycin therapy. Another practical approach would be to define the target concentration by a desired range at 2 h, which corresponds to a Cssmax value of 30-40 mg/L. PMID- 8721275 TI - Pharmacokinetics of moxisylyte in healthy volunteers after intravenous infusion and intracavernous administration with and without a penile tourniquet. AB - The concentration-time profiles of metabolites of moxisylyte (or thymoxamine), an alpha-blocking agent, were investigated in 18 healthy volunteers after intravenous (i.v.) and intracavernous (i.c.) administrations with and without a tourniquet. Four metabolites, unconjugated desacetylmoxisylyte (DAM), DAM glucuronide, and DAM and monodesmethylated DAM (MDAM) sulfates, were found in plasma and urine. For all metabolites, tmax was significantly increased after i.c. administrations and Cmax was significantly decreased. Maximum plasma level of unconjugated DAM was lower after i.c. administration with (1.81-fold) and without (1.26-fold) a tourniquet than after i.v. administration (43.6 +/- 19.6 ng/ml). The elimination half-life of each metabolite showed no change between the three treatments. The difference of 19 min between the mean residence times of unconjugated DAM after i.c. administration with and without a tourniquet may be compared with the difference between the mean duration of the intumescence, that is, 19 min (73 and 54 min with and without a tourniquet, respectively). Total percentages of metabolites recovered in urine were 66.2 +/- 20.9, 61.4 +/- 12.2, and 58.7 +/- 9.1% after i.v. and i.c. administrations with and without a tourniquet, respectively. In conclusion, tourniquet placed before i.c. administration increased the mean residence time of unconjugated DAM of approximately 25% and seemed to increase the efficacy of the drug in healthy volunteers. PMID- 8721277 TI - Changes in vancomycin pharmacokinetics during treatment. AB - Using data gathered in routine monitoring, the pharmacokinetics of vancomycin during the first 10 days of treatment were compared with the pharmacokinetics after 10 days of treatment in 46 adult patients with normal renal function, ages 17-85 years old (mean +/- SD: 50.8 +/- 17.5). The mean time from initiation of treatment to the first sample determination was 5.5 days, and the mean time to the second determination was 13.4 days. Statistical differences between the two periods were observed for all pharmacokinetic parameters, except for the steady state distribution volume. After 10 days of treatment, the mean +/- SD of the vancomycin clearance and elimination rate constant decreased from 1.31 +/- 0.82 to 1.13 +/- 0.72 ml/kg/min (p = 0.0044) and from 0.13 +/- 0.08 to 0.10 +/- 0.06 h 1 (p = 0.091), respectively. The half-life (t1/2) increased from 8.01 +/- 6.82 to 10.02 +/- 8.00 h (p = 0.012). The median percentage of the increment of t1/2 was 9.4%. The increase in t1/2 was > 50% in 12 patients and > 100% in nine cases. No association was found between the increment of t1/2 and the cumulative vancomycin dose. Frequent monitoring of serum vancomycin seems indicated, given the risk of decreased elimination during prolonged treatment. PMID- 8721278 TI - A micromethod for the determination of the new antiepileptic drug levetiracetam (ucb LO59) in serum or plasma by high performance liquid chromatography. AB - An isocratic high performance liquid chromatographic micromethod is described for the quantitation of levetiracetam (ucb L059) in plasma or serum of patients. The chromatography is performed on a 250 x 4 mm I.D. LiChrospher 60 RP-select B, 5 micron column, eluted with an acetonitrile/50 mM phosphate buffer (15:85 vol/vol, pH 5.6) mobile phase, and levetiracetam detected using ultraviolet absorbance at 220 nm. The limit of quantitation was 5 mumol/L and the within-batch and between batch coefficients of variation were < 7%. No interference from commonly prescribed antiepileptic drugs (carbamazepine and its metabolite carbamazepine epoxide, ethosuximide, gabapentin, lamotrigine, phenobarbitone, phenytoin, primidone, valproic acid, and vigabatrin) was observed, and thus the method can be used to monitor levetiracetam in patients on polytherapy antiepileptic drug regimens. PMID- 8721279 TI - Determination of pefloxacin and its main active metabolite in human serum by high performance liquid chromatography. AB - A high-performance liquid chromatographic (HPLC) method is described for the simultaneous determination of a fluoroquinolone, pefloxacin, and its main active metabolite norfloxacin (N-desmethyl metabolite) in serum. Sample preparation involves protein precipitation with acetonitrile. The drugs and the internal standard (acebutolol) were eluted from a 4-microns Novapak C-18 cartridge at ambient temperature with an isocratic mobile phase consisting of 14% acetonitrile in buffer solution, at a flow rate of 2.5 ml/min. The effluent was monitored on a fluorescence detector using excitation and emission wave-lengths of 330 and 440 nm, respectively. Each analysis required no longer than 8 min. Quantification was achieved by measurement of the peak-area ratio of the drugs to the internal standard, and the limit of quantification for both pefloxacin and norfloxacin in serum was 50 ng/ml. The intraday coefficient of variation (CV) ranged from 1.3 to 4.4% and from 2.2 to 7.5% for pefloxacin and norfloxacin, respectively, at the concentration ranges evaluated. The interday CV ranged from 1.1 to 5.9% and from 2.3 to 5.6% for pefloxacin and norfloxacin, respectively, at three concentrations. Relative recovery was 105.5 and 99.5% for pefloxacin and norfloxacin, respectively. Stability tests show that pefloxacin and norfloxacin are stable in serum for at least 3 weeks when stored at -20 degrees C. This method has been used successfully in pharmacokinetic studies in humans. PMID- 8721280 TI - Effects of carbamazepine coadministration on plasma concentrations of trazodone and its active metabolite, m-chlorophenylpiperazine. AB - Effects of carbamazepine coadministration on plasma concentrations of trazodone and its active metabolite, m-chlorophenylpiperazine (m-CPP) were studied in six depressed patients treated with trazodone. The daily dose of trazodone was 150 mg in three cases and 300 mg in three cases. Carbamazepine, 400 mg/day, was coadministered for 4 weeks, and blood samples were taken before carbamazepine addition and at weekly intervals after the addition. Carbamazepine significantly (p < 0.01) decreased plasma concentrations of not only trazodone but also m-CPP at each week. On the average, plasma concentrations of trazodone and m-CPP at 4 weeks were 24 and 40% of the corresponding precarbamazepine values. This study thus suggests that carbamazepine coadministration decreases plasma concentrations of trazodone and m-CPP by inducing the metabolism of these compounds. PMID- 8721281 TI - A systematic review and critical comparison of internal standards for the routine liquid chromatographic assay of amiodarone and desethylamiodarone. AB - Despite potential adverse effects, clinical use of amiodarone is increasing because of its efficacy in treating arrhythmias. Thus there is a continued need for a rapid, practical amiodarone assay to better study the relationship between serum concentrations and clinical effects and to guide safer dosing. Because the most widely used internal standard, L8040, is no longer available, a systematic comparison of potential alternatives was undertaken based on physicochemical and chromatographic characteristics. All amiodarone assays indexed on Medline were reviewed to produce a list of alternatives and five other potential substances considered based on previous experiences. An isocratic high-performance liquid chromatographic method was modified to allow simultaneous resolution of multiple compounds. The internal standard was expected to perform well in the solid-phase extraction of small sample volumes. No commercially available substances were able to duplicate all the advantages of L8040. Tamoxifen, the most acceptable alternative, was used to develop an assay to measure amiodarone and desethylamiodarone at concentrations as low as 0.25 mg/L in 100 microliters of serum (5 ng detected in a 20 microliters injection). Standard curves were linear over the range of concentrations found in our patients (0.25 to 8 mg/L), within run coefficients of variation (CVs) averaged 5.3% for amiodarone and 2.9% for desethylamiodarone, and between-run CVs were 4.5% for amiodarone and 1.6% for desethylamiodarone. PMID- 8721282 TI - A highly sensitive assay for the simultaneous determination of morphine, morphine 3-glucuronide, and morphine-6-glucuronide in human plasma by high-performance liquid chromatography with electrochemical and fluorescence detection. AB - A novel, highly sensitive and specific bioanalytical method has been developed for the simultaneous determination of morphine and its major metabolites, morphine-3-glucuronide and morphine-6-glucuronide, in human plasma, using noroxymorphone as the internal standard. The analytes are isolated from human plasma using a nonpolar/polar C2 solid-phase extraction cartridge and analyzed by high-performance liquid chromatography with serial detection using electrochemical detection for morphine, morphine-6-glucuronide (M6G), and noroxymorphone and fluorescence detection for morphine-3-glucuronide (M3G). The limit of quantitation (sensitivity) using a 0.5-ml sample of plasma is 1 ng/ml, 10 ng/ml, and 5 ng/ml for morphine, M3G, and M6G, respectively. Standard curves were linear (correlation coefficients > 0.999) over the ranges 1-30 ng/ml, 10-500 ng/ml, and 5-100 ng/ml for morphine, M3G, and M6G, respectively. The overall interday accuracy of the method was -1.58% for morphine, 2.27% for M3G, and 5.34% for M6G. The assay is routinely used for the study of morphine, M3G, and M6G pharmacokinetics after oral administration of morphine. PMID- 8721283 TI - Evaluation of a fluorescence polarographic immunoassay with increased sensitivity for measurement of low concentrations of tobramycin in serum. AB - The limits of quantitation of the assay of tobramycin in serum by the fluorescence polarization immunoassay system marketed by Abbott Laboratories (TDxFLx system) are 0.1 and 10.0 mg/L. For some pharmacokinetic studies, however, a more sensitive analysis is needed. The sensitivity of the TDxFLx system can theoretically be increased 10-fold by pipetting buffer solution into the sample well and 450 microliters serum into the predilution well. The assay modified in this way can be run with the usual calibration of the apparatus for normal analysis. The analytical performance of this modification of the TDxFLx assay was assessed. Tobramycin concentrations ranged from 0.01 to 1.0 mg/L. Analytical recovery ranged from 85 to 95%. The coefficients of variation for within-run and between-run precision ranged from 0.5% to 5% and from 2% to 6%, respectively, for control concentrations ranging from 0.05 to 0.75 mg/L. Based on recovery and precision results, the lower limit of quantitation was established as 0.025 mg/L. There was no significant detectable cross-reactivity from ceftazidime, aztreonam, flucloxacillin, cilastatin, or imipenem. There was a small, but significant cross reactivity from gentamicin and netilmicin. Hyperbilirubinemia did not affect the assay, but hyperlipidemia gave falsely elevated results of the tobramycin assay. It was concluded that modification of the assay resulted in an acceptable method to quantify low concentrations of tobramycin in serum. PMID- 8721284 TI - Development of a fluorescence polarization immunoassay for lorazepam quantification. AB - Lorazepam concentrations have been quantified in biological fluids using gas chromatography and high-performance liquid chromatography (HPLC). However, these methods are too time consuming and labor intensive for most nonresearch laboratories to offer. A fluorescence polarization immunoassay (FPIA) method for quantification of lorazepam was developed and validated using a reverse-phase HPLC method as the reference method. The FPIA method involves a single liquid liquid extraction of 100 microliters of either serum or plasma, then direct analysis using the TDxFLx (Abbott Diagnostics, North Chicago, IL). FPIA calibrations were linear between 50 and 800 ng/ml using five calibrators prepared in human serum. Within-run precision (n = 10) for three serum controls (75, 300, and 600 ng/ml) resulted in coefficients of variation (CVs) of 5.7%, 5.4%, and 4.2%, respectively. Between-day precision studies for the three serum controls were 7.0%, 8.9% and 4.9%, respectively (n = 5). The analytical recovery of the three serum controls was 104.9%, 97.3% and 98.3%, respectively. There was an excellent linear correlation between the FPIA and HPLC determinations of 43 patient specimens (r = 0.990, slope = 0.961, intercept = 16.3). No interferences were found from many commonly prescribed nonbenzodiazepine drugs; however, other benzodiazepines that were tested will cross-react with this procedure and give inaccurate results. Therefore, this method should not be used in patients who are receiving other benzodiazepines in addition to lorazepam. The FPIA method described herein can be adapted to reliably measure lorazepam concentrations in serum or plasma. PMID- 8721285 TI - Monitoring of plasma clozapine levels and its metabolites in refractory schizophrenic patients. AB - Plasma concentrations of clozapine and its metabolites desmethylclozapine and clozapine N-oxide were measured in 61 patients with refractory schizophrenia. Before the initiation of clozapine, each patient was given haloperidol (HL) up to 60 mg/day for at least 4 weeks without improvement. Patients were then given a fixed dose of clozapine 400 mg/day. Patients were assessed with the Brief Psychiatric Rating Scale (BPRS) at baseline before HL therapy, at the end of HL at 6 weeks, before clozapine, and after 6 weeks of clozapine therapy. Clozapine and its metabolites were measured by high-performance liquid chromatography with ultraviolet detection. The mean plasma concentrations of clozapine, desmethylclozapine, and clozapine N-oxide were 598 +/- 314, 281 +/- 140, and 90 +/- 29 ng/ml, respectively. The mean decrease in the total BPRS scores from baseline clozapine to the 6-week treatment period was 11 +/- 4. Clinical improvement was noted to occur in most patients with clozapine plasma levels > 300 ng/ml. Improvement diminished in patients with clozapine plasma levels > 700 ng/ml. The most common adverse effects were sedation and hypersalivation. Significant correlations between plasma clozapine concentrations and adverse side effects were not found. PMID- 8721286 TI - Simultaneous determination of lidocaine, bupivacaine, and their two main metabolites using gas chromatography and a nitrogen-phosphorus detector: selection of stationary phase and chromatographic conditions. PMID- 8721287 TI - Stability of FK506 in blood samples. PMID- 8721288 TI - Adolescent suicide. AB - In the introduction to this report our committee, with its focus on adolescent development, expressed its concern that adolescent suicidal behavior represented a grave crisis in the adolescent, a crisis not only in the development of the adolescent but one that endangers the existence of the adolescent. The possibility of a fatal outcome is abhorrent to us as physicians and psychiatrists, as it is to all those entrusted with the care and development of our fellow human beings. Consequently, we explored the ways in which developmental and other forces lead to adolescent suicide and the measures that can be taken to prevent it. We first considered the historical and cross-cultural aspects of suicidal behaviors. Societal and cultural stresses arise from parental attitudes, beliefs, expectations, and childrearing practices that evolve from the social and economic needs in each culture. If unbalanced by growth-sustaining supports, they may compromise or constrict the existential adaptive ability of the developing adolescent and place the adolescent at risk for suicide. Research into vulnerability in adolescence has revealed gender, ethnic, and geographic differences in the dimension of the problem and has indicated the social, psychological, and biological conditions that increase the likelihood that adolescents will resort to suicidal behaviors. Research is still needed to distinguish those adolescents who commit suicide from those adolescents with similar conditions who do not. Research has only begun to explore the ways in which the interaction of specific individual dynamics, precipitating events, and personal characteristics result in an adolescent's attempt of suicide. We discussed the strengths that adolescents acquire, but we emphasized the weaknesses that ensue as adolescents are faced with the impact of the thrust of their own biological, psychological, and social development with the forces inherent in their cultures. Adolescents progress through this period in their lives with varying and varied attempts to master, or cope with, the inevitable change in their existential status. Some try but fail and some fail to try, with resulting despair that can lead those adolescents to believe that suicide is the only choice they have to end their suffering. We described how psychodynamics can influence motivation, relationships, and behaviors, and how these may contribute to an outcome of suicide. Existing psychopathological conditions contribute. These include anxiety, dysthymia, posttraumatic stress disorders, acute reactive disorders, major affective disorders, severe conduct disorders, and psychotic disorders. We considered the possible lethal interplay between psychodynamic and psychopathological factors. This led to the crux of this report, a full discussion of prevention and treatment. The first and most important aspect of suicide prevention is early recognition of the adolescent at risk. It is of high priority to detect and treat those psychiatric disorders accompanied by greatest suicidal risk; depression, conduct disorders, substance abuse disorders, borderline conditions, and schizoaffective disorders. With all adolescents, threats of suicide must be taken seriously. There should be an immediate, complete psychiatric workup preferably before specific treatment begins. If crisis intervention must precede diagnostic study, the workup should not be delayed longer than necessary. Education of health care professionals, educators, families, and peers about warning signs can emphasize early intervention and thereby enable a skilled psychiatrist to assess suicidal thoughts, plans, means, and previous attempts, past and current life stresses, and available family and environmental support. All of this information will lead to a decision regarding hospitalization and treatment for the adolescent.(ABSTRACT TRUNCATED) PMID- 8721289 TI - Healing of muscle trauma after intramuscular injection of antibiotics in sheep: correlations between clinical, macroscopic and microscopic scores. AB - The present study aimed to predict the resultant healing from early lesions (found days 3 and 10 post injection) caused by the intramuscular injection of veterinary antibiotic formulations. Nineteen marketed drugs were selected in order to screen a wide range of irritation conditions at the injection site. Nineteen ewes were each injected intramuscularly with one of the formulations. Each injection was at a different site, 3 and 10 days prior to slaughter. Fourteen of these ewes also received intramuscular injections at two other sites 21 and 32 days prior to slaughter. The tolerance was monitored by clinical examination of the injection site and by gross and microscopic pathology. Myodegeneration and fibre necrosis were determined histologically. The clinical scores did not correlate with the other findings. Myodegeneration correlated with the size of the lesion on day 3 post-injection and was not found thereafter. Although occasionally found alone, it was generally associated with and surrounded by fibre necrosis. When myodegeneration was the only lesion, regeneration was complete by day 21 and the fibrosis was minimal or absent. Necrosis at day 10 post-injection correlated with necrosis at days 3, 21 and 32 post-injection. Fibrosis became prominent around the necrotic muscles from day 10 post-injection. Healing from necrosis was slow with, in some instances, encapsulated debris still persisting at day 32 post-injection. The tissue irritation index correlated well with myodegeneration and necrose (acute lesions) and fibrose and necrose (older lesions). Thus, after considering a large sample of antibiotic formulations, this study indicated that healing could be predicted from the muscle fibre histopathology at days 3 and 10 post-injection. If myodegeneration was found alone, full recovery within 21 days could be predicted. If fibre necrosis was extensive, the healing involved encapsulating the necrotic tissues and thus resulted in extensive scar formation. The tissue changes explained why the irritation index of the lesions at days 21 and 32 post injection could be predicted from their irritation index at days 3 and 10 post injection. Likewise, the size of the lesion at days 21 and 32 post-injection could not be predicted from its size at day 3 post injection. PMID- 8721291 TI - Pharmacokinetic analysis of beta-aminopropionitrile in rabbits. AB - beta-Aminopropionitrile (beta APN), inhibits the activity of lysyl oxidase, an important enzyme for the post-translational formation of inter- and intramolecular covalent cross-linking between the connective tissue proteins, collagen and elastin. We became interested in the possible use of this compound as a therapeutic agent in the so-called human collagen diseases. beta APN's action mechanism is known, but its pharmacokinetics in rabbits have not yet been determined. The present study defined the kinetic parameters of beta APN in rabbits, after oral or intravenous (iv) administration. The HPLC technique was recently modified using OPA (ortho-phthalaldehyde) as the derivative agent. beta APN plasma concentration vs time following the iv administration of 200 mg/kg was best described by the biexponential equation C = 92.43.e(-0.0728 t) + 61.78.e( 0.0088 t) (t1/2 beta = 78.73 +/- 5.19 min; Vc = 1.29 +/- 0.04 L.kg-1). After oral administration, beta APN followed a zero-order absorption pattern (Ko = 3.02 +/- 0.34 mg.kg-1.min-1), which means that the beta APN reached the blood very quickly. PMID- 8721290 TI - The proliferative responses of cow stripping milk and blood lymphocytes to pokeweed mitogen and ginseng in vitro. AB - Milk samples were collected directly after the morning milking (stripping milk) from eight cows of the Swedish Red and White breed, during the period of mid lactation. The milk cells (29 +/- 11 x 10(6)) were obtained after centrifugation and by passing the cell pellet through a cotton column. They consisted of 36 +/- 9% lymphocytes, 14 +/- 2% macrophages and 50 +/- 10% polymorphonuclear leukocytes. The autologous peripheral blood lymphocytes were separated by means of a Ficoll-gradient. The lymphocytes from the blood and milk were subjected to the lymphocyte stimulation test (LST). The stimulation indices (SI) demonstrated that pokeweed mitogen (PWM) induced a greater degree of proliferation in the autologous peripheral blood lymphocytes than it did in the stripping milk lymphocytes (39 +/- 8 vs 5 +/- 1, respectively, P < 0.01). Ginseng (GS), the root of Panax ginseng CA Meyer, has been used in China for thousands of years to enhance the body's resistance to many diseases. In the present study, a dry ginseng extract was investigated to evaluate its immunomodulating effects in vitro on the peripheral blood and stripping milk lymphocytes. The response was significantly augmented (approximately 20%) in the peripheral blood lymphocytes incubated with GS extract in combination with PWM, as compared with the cells incubated with PWM alone. Increased responses were also detected with the lymphocytes from stripping milk incubated with GS+PWM, but to a much lesser extent (approximately 9%) compared with the autologous peripheral blood lymphocytes. PMID- 8721292 TI - Changes in inflammation-related blood constituents of mastitic cows. AB - The following blood constituents were measured in the blood and/or serum from healthy and mastitic cows: lipid peroxides (LPO), alkaline phosphatase (AP), lactate dehydrogenase (LDH), urea, glutathione peroxidase (GSH-Px), prostaglandin E2 (PGE2), cyclic AMP (cAMP), electrolytes (Na, K), white blood cells (WBC), haemoglobin (Hb), haematocrit, eosinophils, and electrical conductivity. The levels of these blood variables, excluding GSH-Px, erythrocytes variables and conductivity, were higher in mastitic than in healthy animals. In the blood of the mastitic animals, the AP and LPO levels were 44 and 38% higher than in the healthy animals. The substantial decline in the GSH-Px levels in mastitic animals (P < 0.05) may be related to changes in lipid peroxidation and PG formation and possibly to oxidative stress caused by the host defences. The eosinophil levels were positively correlated with those of PGE2 and cAMP. The possible implications of these findings for the diagnosis of mastitis are discussed. PMID- 8721293 TI - [Bioavailability of muscle creatine kinase in sheep. Application to the assessment of local tolerance to injectable veterinary formulations]. AB - Pharmacokinetic variables of skeletal muscle creatine kinase were determined in sheep after intravenous and intramuscular administration of the semipurified enzyme. Catheters implanted in the jugular vein were used for both intravenous injections and blood withdrawals. Blood sample collection by vacutainer and hemolysis may in fact have considerable effects on the measurement of creatine kinase activity in plasma. The change in the enzyme activity versus time in the plasma, after intravenous administration (123 +/- 38 U/kg) of creatine kinase, was fitted by a biexponential model. The mean volume of the central compartment (45 +/- 5 mL/kg) was approximately equal to the plasma volume. Plasma half-life and plasma clearance of creatine kinase were 3.7 +/- 1.7 h and 23 +/- 8 mL.kg-1.h 1, respectively. Mean plasma bioavailability of creatine kinase after intramuscular administration (357 +/- 36 U/kg) in both the loins and the gluteal mass was 42%. Maximal plasma activity was observed 4 and 5 h after injection and the half-life of the terminal phase was 7.3 or 8.6 h according to the muscle. The extent of muscle damage after intramuscular administrations of 21 veterinary drug formulations (one product per animal) was estimated from the total creatine kinase activity released in plasma during the 72 h following the injection. Equivalent weights of damaged muscle ranged from 1.4 to 83.3 g according to the irritant potency of the test formulation. Results differed only moderately between the injection sites (right and left gluteal mass) in the same animal. It can be concluded from this study that, in sheep: i) the bioavailability of creatine kinase from different injection sites (gluteal mass and loins) is comparable; and ii) the intra-individual variability in the estimation of muscle damage is moderate. Once validated, this non-invasive approach for local tolerance studies could be of value in assessing and comparing the irritant potency of veterinary drugs and in reducing the number of animals required. PMID- 8721294 TI - Pharmacokinetics of penicillin G in plasma and interstitial fluid collected with dialysis fiber bundles in sheep. AB - A new method of collection of interstitial fluid (ISF) (the site of most bacterial infections) was developed for the determination of free (unbound) penicillin G concentrations in sheep. Dialysis fiber bundles for the collection of ISF were first characterized in vitro and subsequently implanted in the subcutaneous fascia of the dorsal thorax parallel to the vertebral column in sheep. The sheep were then dosed intravenously with 26.4 and 52.9 mg/kg of sodium penicillin G using a crossover experimental design. Plasma and ISF dialysate were collected after dosing for determination of penicillin G concentrations using high pressure liquid chromatography (HPLC). The concentration of penicillin G in the ISF dialysate was calculated with the recovery ratio determined for each fiber bundle. The decline of penicillin G concentrations in ISF dialysate paralleled the disappearance of the drug from plasma providing evidence for the rapid diffusion of penicillin G into the fiber bundles. Pharmacokinetic analysis determined that the disposition of penicillin G was best described by a two compartment open model with penicillin concentrations in plasma (Cp) defined by two biexponential equations, Cp = 170.64e-7.16t + 31.04e-1.56t for the low dose and Cp = 418.19e-1.56t for the high dose. PMID- 8721295 TI - Are stable flies (Diptera: Stomoxyinae) vectors of Trypanosoma vivax in the Central African Republic? AB - The epidemiology of Trypanosoma vivax infections was studied at a riverside site in the Ouro-Djafoun livestock area situated in the Central African Republic during the period between July 1991 and July 1992. This paper examines the possibility that stable flies (Diptera: Stomoxyinae) were also vectors of this trypanosome species in a non-cyclic way. Previous studies have revealed that the usual cyclic transmission by the tsetse fly Glossina fuscipes fuscipes was probably not the only transmission route. At the study site, at least five species or subspecies of stable flies were encountered: Stomoxys nigra nigra (approximately 60% of the sample), S taeniata, S sitiens, S omega omega and Haematobia spp. The hypothesis that stable flies could be good vectors of T vivax in this country is supported by three main observations: i) stable flies were very abundant at the cattle resting site; ii) an estimation of the 'contact index' between the cattle and stable flies demonstrated close interactions between cattle and stable flies at this site, particularly during the rainy season, and iii) there was a good correlation (P < 0.05) between the apparent stable fly densities at the resting site and the frequency of T vivax in the cattle. The relevance of this phenomenon in terms of epidemiology and combatting T vivax-caused nagana is discussed. PMID- 8721296 TI - Caprine and bovine B rotaviruses in western France: group identification by Northern hybridization. AB - In a survey of coronavirus and rotavirus-induced neonatal diarrhoea in 373 calves from 284 farms, nine (2.4%) of the animals were found to be infected with non group A rotaviruses when their faeces were analysed in a commercial ELISA and by PAGE. Seven out of eight 1-4- day-old kids from a single farm were also infected with similar viruses. In a mini-PAGE, all the viruses displayed group B- and/or E like 4223 electrophoretypes. In Northern hybridizations with cDNA chemiluminescent probes specific to rotavirus groups A (RF strain), B (adult diarrhoea rotavirus, ADRV) and C (Cowden strain) all the viruses belonged to group B. PMID- 8721298 TI - Assessment of the conventional detection of fecal Cryptosporidium serpentis oocysts in subclinically infected captive snakes. AB - Fecal specimens of seven captive snakes naturally subclinically infected with Cryptosporidium serpentis were monitored for 131 days for the presence and concentration of oocysts. Thirty-three of 81 (41%) of the monoclonal antibody positive stools were read as negative (sensitivity 59%) by the acid-fast-stained (AFS) fecal smears. Oocyst concentrations in the false-negative stools ranged between 6.0 x 10(3)/g to 2.5 x 10(4)/g. The experimentally determined concentration-threshold of oocyst detection by AFS fecal smear was 3.0 x 10(4)/g. The stool weights did not conform to a normal distribution; the oocyst concentration was significantly correlated with the stool weight (P < 0.01). Due to this correlation, stools which constituted less than 0.41% of the snake weight were classified as negative by the AFS fecal smears. The AFS fecal smear technique should be used exclusively for the determination of Cryptosporidium positive snakes, but not for diagnosis of snake negativity for Cryptosporidium; it should be applied only for larger fecal specimens, such as those which constitute more than 0.41% of snake weight. At least five to seven stool samples should be examined by fecal smear in order to determine snake positivity for Cryptosporidium in subclinical infection. PMID- 8721297 TI - Uterine estrogen and progesterone receptors in prepubertal ewes: distribution in myometrium, endometrium and caruncles. AB - In order to study the expression of uterine steroid receptors during animal development, estrogen and progesterone binding sites (E2R and P4R) were investigated in 9-month-old Corriedale ewe-lambs during the non-breeding season. E2R and P4R were identified by binding assays and Scatchard analysis in all uterine layers. The dissociation constant (Kd) values were: 0.63 +/- 0.14 nM (n = 24) for E2R and 2.10 +/- 0.32 nM (n = 24) for P4R. The E2R distribution was: 188 +/- 32, 367 +/- 83 (NS) and 494 +/- 72 fmol/mg protein (P < 0.02) for myometrium, caruncles and intercaruncular endometrium, respectively. Interestingly, high levels of P4R (fmol/mg protein) were found. By layer the P4R content was: caruncles, 1 123 +/- 198; intercaruncular endometrium, 1 283 +/- 187 (NS); and myometrium, 502 +/- 76 (P < 0.002). A positive correlation existed between both receptors (r = 0.772, P < 0.0001, n = 24). E2 and P4 circulating levels (measured by radioimmunoassay), were similar to the adult ewe basal levels. These findings suggest that ewes have sensitive and functional uterine estrogen and progesterone receptors even before cyclic ovarian activity. PMID- 8721299 TI - Urine concentrations in the spaces of the sheep renal pelvis. AB - Urine concentrations were measured in samples from the ureter, the pelvic cavity, close to the tip of the papilla, and from the most proximal part of the fornices of sheep kidneys. It was found that the concentration of urea, the osmolality, and the urine-to-plasma ratios of inulin, urea and the osmolality were higher in the ureter than in the pelvic cavity. The same parameters were found to be higher in the pelvic cavity than in the fornices except for the urea concentration which was unchanged. The results of this study revealed the various concentrations of urine in the ureter, the pelvic cavity and the proximal part of fornices of sheep kidney. PMID- 8721300 TI - [Concentrations of metabolically relevant parameters in the blood of highly pregnant ewes]. AB - From 38 healthy ewes (20 nonpregnant, 18 pregnant) of different flocks and two races (Blackhead Mutton and Merino sheep) concentrations of glucose, ketones, triglycerides, cholesterine and urea in blood plasma were determined to prove if there were any differences between pregnant ewes in the last nine days before parturition and nonpregnant ones. In nonpregnant ewes plasma values of glucose were significantly higher than those of clinical inapparent pregnant ones with twins in the last four days before parturition. Blood values of beta hydroxybutyrate and cholesterine, partly of triglycerides too, were significantly higher in late pregnant ewes with one or two fetuses. In contrast, concentrations of alpha-ketoglutarate and urea showed no significant differences between both groups, pregnant and nonpregnant ewes. The determination of blood plasma concentrations of beta-hydroxybutyrate, triglyceride and cholesterine, as well as glucose in the case of twin-pregnancy, can provide an important information to estimate the animal's state of metabolism in late pregnancy. PMID- 8721301 TI - [Neurohormonal control of gallbladder motility by intra-ileal and intracolonic nutrients--a review]. AB - The duodenum controls gallbladder motility mainly via stimulatory mechanisms, whereas intraileal and intracolonic nutrients have mainly inhibitory effects on postprandial as well as interdigestive gallbladder motility, which are described in particular. Possible mechanisms for the neurohormonal mediation of the effects of intraileal and intracolonic nutrients on gallbladder motility as well as the possible physiological importance of these effects are discussed. PMID- 8721302 TI - [Dust emission from chicken and layer houses]. AB - Dust concentration was measured by Laser-monitor in two at a time houses for layers with litter or with cages and in two at a time houses for growing chicken with litter or with cages in the middle of the keeping area, in the outlet opening and in 3, 10, 50 and 100 m distance over a period of 8 weeks in winter, summer and spring/autumn. Dust values have shown only less dynamics between 8 am and 5 pm. Significant positive correlation was found between dust concentration and age of poultry, keeping time resp. in litter housing system but not in cage housing system. Also, it was demonstrated a significant relation between dust concentration inside of poultry house, at the outlet openings and in 3 m distance in the outdoor air. There was no influence evident of indoor dust concentration on the dust value in the environment in more than 3 m distance to the poultry house. It was shown a low dust concentration of 0.1% up to 2.1%--compared to the indoor dust values--in 50 and 100 m distance to outlet ventilation system independent on housing, climatic and topographic factors. PMID- 8721303 TI - [Demonstration of pneumonia in swine as a constant problem: culture and immunofluorescence microscopic studies of bronchioalveolar lavage (BAL) and serological findings]. AB - Bronchoalveolar lavage was performed in 128 pigs from five fattening units showing acute pneumonia (48 animals), subclinical purulent pneumonia (17 animals), and chronic purulent pneumonia (63 animals). These samples were investigated for bacteria. Additionally immunofluorescence microscopy as well as serological investigations were performed to detect antibodies against several bacteria and viruses. Pasteurella multocida could be detected in more than a half of the samples of pigs with acute pneumonia. Bordetella bronchiseptica and mycoplasmas were isolated in a lower amount. Probably these bacteria infected the pigs of at least one herd after a primary infection with influenza virus because (i) influenza virus could be detected in three of four animals investigated for influenza virus by culture methods, (ii) the virus could be detected in one third of the animals investigated for by immunofluorescence microscopy, and (iii) antibodies against influenza virus could be detected in almost all animals. From pigs with subclinical purulent pneumonia Bordetella bronchiseptica as the only bacterial lung pathogen could be isolated exclusively from nearly each sample. From the samples of pig suffering from chronic purulent pneumonia first of all Bordetella bronchiseptica, Pasteurella multocida and different mycoplasma species could be detected. Using cultural methods Actinobacillus pleuropneumoniae could be isolated from six samples only, in contrast to frequent positive reactions against Actinobacillus pleuropneumoniae antigens obtained by immunofluorescence microscopy and CFT. PMID- 8721304 TI - [Management of E. coli dependent factorial diseases in weaning piglets]. AB - The most important postweaning factorial diseases are at least partly caused by E. coli. The term postweaning coli complex can be subcategorized into the following manifestations: postweaning diarrhoea, edema disease, postweaning wasting and hemorrhagic gastroenteritis. In the presented study the effect of prophylactic zootechnique alone and zoo- and biotechnique in combination was evaluated during the first weeks postweaning. The results showed that combined zoo- and biotechnique is superior to simple zootechnique regarding food conversion (1.41 kg versus 1.73 kg), average daily weight gain (390 g versus 325 g) and postweaning piglet mortality (3.1% versus 4.9%). It is the opinion of the authors that combined postweaning zoo- and biotechnique should be performed in such pig production units where ETEC and/or SLTEC are present. PMID- 8721305 TI - Biomimetic-dye affinity chromatography for the purification of mitochondrial L malate dehydrogenase from bovine heart. AB - Seven biomimetic anthraquinone triazinyl dye-ligands, bearing as triazine-linked terminal moiety (keto)carboxylated structures mimicking substrates and inhibitors of malate dehydrogenase (MDH), were immobilised on cross-linked agarose Ultrogel A6R. These biomimetic ligands are terminal-ring analogues of commercial nonbiomimetic Cibacron blue 3GA (CB3GA) and parent Vilmafix blue A-R (VBAR). The biomimetic-dye adsorbents, along with nonbiomimetic adsorbents bearing immobilised CB3GA and VBAR, were evaluated for their ability to purify mitochondrial malate dehydrogenase (mMDH) from bovine heart. All but two biomimetic-dye adsorbents displayed higher purifying ability for MDH, compared to nonbiomimetic-dye adsorbents. Furthermore, immobilised anthraquinone-dyes were able to discriminate between the mitochondrial and the cytoplasmic MDH isoenzymes, binding only to the former. One immobilised biomimetic-dye (BM5), bearing as biomimetic terminal moiety 4-aminophenyloxanylic acid, showed the highest purifying ability. This affinity adsorbent was exploited in the purification of mMDH from unpretreated bovine heart extract in one-step. The procedure afforded mMDH at 54% overall yield and of specific activity approx. 1300 U mg-1 (25 degrees C), using step-elution with a mixture containing 0.1 mM beta-nicotinamide adenine dinucleotide (NAD+) and 1.5 mM sulphite. Commercial analytical-grade bovine heart mitochondrial MDH, when assayed under identical conditions, gave a specific activity not exceeding 950 U mg-1. The well-known adsorbent Cibacron blue 3GA-agarose exhibited 8% lower recovery and 25% lower purification for mMDH. The product obtained from the procedure based on the BM5 adsorbent was free of cytoplasmic MDH, glutamic-oxaloacetic transaminase (GOT) and fumarase, and since it has also shown high specific activity, it should be suitable for analytical applications. PMID- 8721306 TI - Overexpression and purification of human immunodeficiency virus type 1 env derived epitopes in Escherichia coli. AB - In order to develop a reliable and inexpensive serodiagnostic method, a part of envelope gene of HIV-1, gp120' and gp41' (HIV-1 env a.a. 295-474 and a.a. 556 647) was cloned into a T7 expression vector (pET3d). The fusion protein (gp120' gp41') was overexpressed in Escherichia coli, then purified to homogeneity by a simple gel filtration chromatography. Western blot analysis and enzyme-linked immunosorbent assay (ELISA) using the purified fusion protein showed a high sensitivity and a specificity for the detection of anti HIV-1 antibodies in testing human plasma. These results suggest that the expression scheme employing a direct expression vector and the rapid purification method are reliable and applicable for obtaining a large quantity of HIV-1 env protein for diagnoses of HIV-1 infections. PMID- 8721307 TI - Investigating the use of the chymosin-sensitive sequence of kappa-casein as a cleavable linker site in fusion proteins. AB - The chymosin-sensitive sequence of bovine k-casein A (kappa-CN A) was investigated as a cleavable linker site between the two domains of a streptavidin chloramphenicol acetyltransferase fusion protein. Two DNA sequences were synthesized which encode the amino acids from 101 to 107 and from 97 to 113 of bovine kappa-CN A. These sequences were separately cloned in-frame to a streptavidin expression vector used for fusion protein construction. The gene for chloramphenicol acetyltransferase (CAT) was then cloned in-frame to a streptavidin-chymosin-sensitive linker vector forming plasmids pStCL1CAT and pStCL2CAT. The fusion protein was expressed in Escherichia coli and SDS-PAGE and Western blot analysis of chymosin-treated cell lysates showed a pH-dependent cleavage of the fusion proteins. Fusion proteins were also bioselectively immobilized onto biotinylated controlled-pore glass beads and treated with chymosin. CAT was specifically released by chymosin treatment and was identified by SDS-PAGE. PMID- 8721308 TI - [A model of the spatial organization of double-row (pseudostratified) epithelium (a topical article)]. PMID- 8721309 TI - [A comparative study of carboxylesterase in the white mouse and in the caterpillar of the cotton bollworm Heliothes armigera]. AB - Studies have been made on enzymic hydrolysis of p-nitrophenylacetate (p-NPhAc), n nitrophenylbutyrate (p-NPhBu) and indophenylacetate (IPhAc) by carboxylesterase (CE) from mouse blood plasma and liver as well as from caterpillar of the cotton worm haemolymph, intestine and fat body. Different KM and V max values were obtained for CE from these sources. The highest specific activity of CE from mouse liver and caterpillar intestine and fat body was observed with p-NPhBu. p NPhAc is the best substrate for CE from mouse blood serum and caterpillar haemolymph. Carboxylesterases from mouse and caterpillar differed in their sensitivity to armine and paraoxone by 1-2 orders depending on the substrate used. Species and tissue differences in the kinetics of CE-catalyzed reactions with different substrates and inhibitors were revealed. PMID- 8721310 TI - [The role of the central substantia grisea of the midbrain in regulating the sound emission process in the bat Rhinolophus ferrumequinum]. AB - Using axonal transport of HRP, relatively small region of the lateral part of the central gray matter of the midbrain was revealed the neurones of which send afferents to motoneurones of the vagal nucleus. It was shown that practically complete damage of the central gray matter does not affect temporal and frequency parameters of echo-sounding signals produced by animals during total location, do es not affect complex mechanism of stabilization of the filling frequency of these signals, as well as does not affect the ability of animals to decrease the filling frequency in relation to the rate of approaching the target. PMID- 8721312 TI - [The comparative characteristics of the physicochemical and structural-functional properties of hemoglobin in a number of vertebrates]. AB - Using isoelectrofocusing within pH gradient 6-8, studies have been made on microheterogeneity of haemoglobins from various vertebrates. Phylogenetic evaluation of oxygen affinity of the main and minor electrophoretically pure haemoglobin fractions is given. It was shown that oxygen affinity of the haemoglobins is related to the ratio between dicarboxylic and basic amino acids. PMID- 8721311 TI - [The sex and corticosteroid hormones of rats selected for estrous-cycle reaction to constant illumination]. AB - Two substrains of rats selected for the high (ESTH) and low (ESTL) ability to develop permanent oestrus under constant illumination were produced. The concentration of estradiol, testosterone in blood and 11-OCS in blood and adrenals was studied in rats of the 29 th generation under normal day (14L : 10D) and constant illumination (CL). The concentration of estradiol in blood of control rats in proestrus was 61.6 +/- 4.1 pg/ml and did not differ from substrain ESTH (59.6 +/- 4.5 pg/ml) and ESTL (58.8 +/- 4.8 pg/ml); in oestrus, the minimal concentration of estradiol was noted in control-(46.5 +/- 3.1 pg/ml), maximal one-in rats of substrain ESTL (63.3 +/- 6.2 pg/ml), in ESTH-55.8 +/- 1.7 pg/ml. The maintenance of rats in condition of CL brought about the tendency to decrease the level of estradiol in blood of rats selected in proestrus and to increase it in oestrus in comparison to control. The level of testosterone in blood of control rats in proestrus-oestrus changed in parallel to concentration of estradiol. The concentration of testosterone in blood of selected rats was higher in proestrus-oestrus as compared to control. In condition of CL, the testosterone level in proestrus did not differ from control, in oestrus it was higher than in control. The level of 11-OCS in adrenals of selected substrain was higher than in control as well in condition of 14L : 10D as in CL. The exception was substrain ESTH-in proestrus the concentration of 11-OCS in condition of CL was lower than in control. CL drastically changed direction of cyclic shifts in level of 11-OCS from higher value in proestrus under condition of 14L : 10D to low one in condition of CL. The fluctuation of 11-OCS level in blood in groups of experimental rats usually was parallel to that in adrenals. PMID- 8721314 TI - [ACTH and the processes of higher nervous activity in the hedgehog Erinaceus auritus]. AB - Using feeding behaviour model in hedgehogs, studies have been made of the effect of ACTH1-39 on learning, memory and the pre-formed conditioned reactions. It was shown that preliminary administration of the hormone produces facilitating effect on learning. In animals with pre-formed conditioned reflexes ACTH decreases temporal parameters of these reflexes, increasing intersignal and locomotor activity. Injection of ACTH favours memory processes. Intensification of trace conditioned reactions takes place at the background of hyperphagia, vigorous motor activity, etc. PMID- 8721313 TI - [A comparative study of the intracellular regulation of transport ATPase activity in non-nucleated erythrocytes]. AB - Removing the basic cytoskeleton proteins (spectrin, 4.1 and 2.1 band proteins) from the unnuclear erythrocyte membranes of four mammalian species (mouse, hamster, guinea, pig and rabbit) made no changes in Na,K-ATPases, while the specific activity of Ca-ATPases was decreased (the decreasing was of a range of 60-90% depending on the species, in values per mg of protein). The specific activity of the Na,K-ATPase (per ml of cells) varied from 20 to 30% in different species. Incubation of ghosts with erythrocyte haemolysates from rat led to a markedly expressed activation of the Na,K-ATPase in in erythrocytes of all species studied, while the incubation with erythrocyte haemolysate of the same species caused activation of the enzyme in mouse and guinea pig ghosts only. The data obtained support a concept of intracellular Na,K-ATPase activator (probably a protein) having no absolute species specificity. They also show that normal transport function of ATPases in unnuclear erythrocytes needs integrity of the membrane skeleton. PMID- 8721315 TI - [The discrimination by the long-tailed Arctic suslik Citellus parryi of short frequency-modulated sounds]. AB - Using conditioned reflex technique, ground squirrels have been examined for their ability to discriminate between species specific FM-shaped signals and synthetic sounds. The animals reliably distinguish a complex FM-signal from gradually sweeping downwards sounds only when they differed either in their duration or in steepness of FM sweep. PMID- 8721316 TI - [Central hemodynamics based on rheoencephalographic data in elevated arterial pressure in man]. PMID- 8721317 TI - [The effect of hemolysate and calcium ions on transport ATPase activity in guinea pig erythrocytes]. AB - Influence of 0.1, 0.3 and 1.0 microM Ca++ on Na,K-ATPases in different membrane preparations of guinea pig erythrocytes was studied both in presence and in absence of haemolysate. The incubation with the haemolysate and 0.1 or 0.3 microM Ca++ increased significantly the activity of the Na,K-ATPase in all membrane preparations used. The increasing Ca++ concentrations (up to 1 microM) led to the loss of the activating effect of the haemolysate in membrane preparations with lack of a part of the cytoskeleton proteins, and also decreased sharply its activity in all the preparations in absence of haemolysate. The latter also increased the Ca-ATPase activity irrespectively of the integrity of cytoskeleton structures (may be because of calmodulin action). The results obtained suggest that the cytoskeleton structures mediate the intracellular Na,K-ATPase activator action. PMID- 8721318 TI - [The biological action of physical factors in the critical periods of embryogenesis]. AB - The critical period in a chicken embryonic development (the 10-13-th days of incubation) is revealed under total electromagnetic radiation. This external factor is a physiologically active irritant which can influence functional state of the brain. The increased absorption of electromagnetic energy takes place in this incubation period. Its dynamics within 20 days of embryonic development has phasic, up and down character. Electromagnetic exposure (4 hours a day daily) in the above mentioned period evokes a delay in embryo adaptive motor behavior (biofeedback learning). Morphological investigation shows significant pathological changes--destruction of share brain synapses. The delay in embryo hatching for a day is also detected. Radiation exposure within other periods of incubation (3-6-th or 12-15-th days) was not effective with respect to formation of normal motor pattern in biofeedback experiment. PMID- 8721319 TI - [The effect of changes in the level of interoceptive afferentation on the spontaneous periodic motor activity of rat pups]. AB - The effect of intraperitoneally injected (25 mg/kg) novocaine (procaine) on the spontaneous periodic motor activity was investigated in rat puppies ageing from 1 to 14 days. It was shown that reactions were phasic: initial continuous and then elevated activity was followed by depression and recovery. The duration and intensity of stimulation and depression decrease with age. It was concluded that in newborn animals an excitability of motor centers depends on the level of interoceptive afferentation. The last may be one of the motor activity regulation mechanisms. The role of this mechanism decreases with age in favour of extero- and proprioception mechanisms of motor control. PMID- 8721320 TI - [The afferent and efferent connections of field 7 in the parietal cortex with field 18a of the visual cortex in the rat]. AB - By means of administration of horseradish peroxidase to the field 7 in the parietal cortex of rat, studies have been made on antero- and retrograde axonal transport of the enzyme to reveal bilateral connections of this field with the field 18a of the visual cortex. It was shown that the field 18a is a source of afferent fibers which reach the field 7, more evident projections being observed at the ipsilateral side. Labeled neurones in the field 18a were found mostly in layer III, and to a lesser extent-in layer V. Single labeled neurones were also found in layers II and VI. Labeled terminals of efferent fibers originating from cells in the field 7 were located in layers I, II-III and V of the field 18a. Maximum density of the terminals was registered in layer II-III, less one in layer V, and very negligible-in layer I. It should be noted that labeled terminals were revealed in those cortical regions, which contained labeled neurones which gave origin to afferent fibers to the field 7. PMID- 8721321 TI - [The modelling of different forms of natural selection in cells of the ciliated epithelium in the frog Rana temporaria]. AB - By means of thermal and non-thermal selection of parental gametes in the frog Rana temporaria, the effects of normalizing and canalizing forms of natural selection have been reproduced on the ciliary epithelium of the progeny. It was shown that the heat resistance level of epithelial cells in the progeny is inherited from both parents. PMID- 8721322 TI - [Homeotic genes and the origin of syntax]. PMID- 8721323 TI - [Ethological investigations in various housing of broiler chickens]. AB - Ethological observations showed an increase of moving activity, comfort and social behaviour as well as improvement of resting behaviour when stock density is reduced and the light program contains splitted phases of darkness. PMID- 8721324 TI - [Performance of broiler chickens and quality of litter in various housing conditions (stocking density)]. AB - In the present study the influence of a reduced stocking density on body weight, feed and water intake as well as the dry matter and nitrogen-centent (N) of bedding was examined in 3 fattening periods (2 x conventional closed stable, each 36 days, 1 x Louisiana stable, 40 days). In the controlls (stocking density 38 kg/m2) the broilers achieved a mean slaughter weight of 1497, 1411 and 1681 g, under experimental conditions (stocking density on average 33 kg/m2) the compareable figures were 1555, 1431 and 1793 g. Feed conversion ratios were better and the mortality during the fattening periods were lower in groups with reduced stocking density than in the controlls. There was no remarkable improvement of litter quality during the fattening periods in dependence on reduced stocking density. The dry matter content of litter decreased to 60%. At end of fattening the bedding contained 6-9g NH3-N, 7-17 g urea-N and 0,4-4 g uric acid-N/kg wet weight. The lowest uric acid values were found in the bedding samples from Louisiana stables. Effects of a reduced stocking density became obvious only in conventional closed stables by lowered NH3-N-concentrations. The stocking density had no influence on the distribution of the different fractions of NPN with regard to total N. PMID- 8721325 TI - [Evaluation of different broiler housing conditions on the basis of clinical, pathological, anatomical and laboratory diagnostic criteria--a summary]. AB - In a cooperative study, the impact of different broiler housing conditions (see communication BUCHENAUER) on clinical, post-mortem and laboratory diagnostic (microbiological, parasitological, serological, immunological) parameters was investigated in the course of three different fattening period. Clinically, it was observed, that lower stocking densities allowed a relatively higher motility of the birds. On the other hand, necropsy findings, microbiological, parasitological, serological and immunological findings gave no indications that these parameters were influenced statistically significant by either one of the broiler keeping systems. PMID- 8721326 TI - [Animal housing climatic quality in various harmful conditions in broiler production to avoid heat stress in summer]. AB - There is an ongoing debate about the optimum stocking density of broiler birds. The figures which are discussed range between 25 kg/m2 and 43 kg/m2. The Ministry for Agriculture, Forestries and Food of Lower Saxony issued a decree which limits the stocking density to 30 kg/m2 in winter and 27 kg/m2 in summer time. Among other rules a day-night-rhythm in lighting has to be installed and the maximum allowable ammonia concentration is limited to 20 ml/m3. In a field study, involving 5 institutions of the Hannover School of Veterinary Medicine and the University of Gottingen, the effects of the decree on animal behaviour, health, hygienic conditions and economy was investigated in two identical conventional broiler barns in winter (trial 1) and summer (trial 3) and in two identical louisiana-type barns in summer (trial 2). This paper reports on the comparison of the air quality parameters in the experimental barns (E) and control barns (C) and concentrates on mean values only. The following differences were observed. (1) The mean air temperatures between the E-barns and C-barns didn't differ by more than 1 degree C. (2) The mean relative humidity of the air was in barn E 3.6% lower than in barn C (trial 1), the differences of the mean values in the other two trials were below 1% r. h. (3) The mean ammonia (NH3) concentrations were generally higher in the E-barns (trial 1: E = 17.9, C = 13.4, trial 2: E = 14.5, C = 10.9, trial 3: E = 7, C = 4.8 ml/m3) than in the C-barns. The NH3 threshold of 20 ml/m3 was frequently exceeded in trial 1 and 2 reaching peaks up to 46 ml/m3. (4) No hazardous concentrations of carbon dioxide (CO2), nitrous oxide (N2O), dimethyl amine (HN(CH3)2) and methane (CH4) were observed. The highest CO2 concentrations were found in trial 1 barn C (2816 ml/m3), 300 ml/m3 more than in barn E. In trials 2 and 3 the CO2 concentrations were slightly higher (trial 2: 200, trial 3: 40 ml/m3) in the C-barns than in the E-barns. The highest N2O concentrations were found in the louisiana-type barns (trials 2: E = 2.3, C = 1.8 ml/m3). In trial 1 (E = 0.5, C = 0.6 ml/m3) and trial 3 (E = 0.2, C = 0.2 ml/m3) the mean concentrations were by factors 3 to 10 lower. Small amounts of HN(CH3)2 were found in trial 1 only (E = 0.2, C = 0.3 ml/m3). The mean CH4 concentrations were higher in the C-barns (trial 2: E = 12.6, C = 14.6, trial 3: E = 19, C = 19.6 ml/m3). (5) The mean inspirable dust concentrations didn't differ very much between the trials an the barns. Highest inspirable dust amounts were found in trial 1 (E = 3.3, C = 3.8 mg/m3) followed by trial 2 (E = 2.8, C = 2.7 mg/m3) and trial 3 (E = 2.5, C = 2.3 mg/m3). A similar tendency was seen in the respirable dust (trial 1: E = 0.4, C = 0.6, trial 2: E = 0.4, C = 0.4, trial 3: E = 0.4, C = 0.3 mg/m3). The results of this study indicate that a reduction of bird density to 30 kg live weight per m2 will not have an immediate and strong effect on improving air quality. In particular heat stress in summer remains a problem. Therefore a brief list of measures to reduce heat stress for broilers in included. PMID- 8721328 TI - [The African ostrich--a "useful animal" in Germany?]. AB - Farming of the ostrich--a non domesticated species--cannot be done species conforming; it is even a cruelty to the animals. Keeping in captivity leads to a steady rank order stress; the biology of their reproduction is heavily impaired and their typical moving requirements can only be poorly satisfied. The specific habit of food intake cannot be performed accordingly, and the early deprivation, caused by the absence of parents during the weanling period, creates extremely abnormal behaviour. Summing up, all these deficiencies constitute the fact of cruelty to animals according to the German Animal Protection Act. PMID- 8721327 TI - [Results of pathological-anatomical studies of limbs and spinal column]. AB - The pathomorphological study was carried out on a total of 360 broiler chicken which had been random-sampled on days 22 and 35 of the mast period, respectively, from those flocks that were to be compared. Furthermore, 161 animals with evidence of movement disturbance that were slaughtered in the last two mast weeks, were also evaluated. With regard to incidence, severity of movement disturbance and the spectrum of pathomorphological changes of the skeleton there were no differences between the different groups. When histological and morphometrical methods were applied, no differences in the skeleton structure were noted between flocks with conventional housing and flocks with reduced population density. Equally, no differences were ascertained with regard to dry substance and ash content of the bones including the minerals calcium and phosphorus. Furthermore, there were no group differences in serum calcium, phosphorus and alkaline phosphatase. In numerous chicken from all groups a plantar pododermatitis with differing incidence and strongly varying intensity was observed. The lesions were characterized by a papillomatous proliferation of the basal epithelial layers. There was widespread inflammation with loss of the epithelial layer and deep necroses of the sole. A latent infection with papilloma viruses is discussed. About 90% of the random-sampled chicken of all groups showed bending of the vertebral column by 20 degrees at the height of the 6th thoracic vertebra. In numerous chicken the 6th thoracic vertebra was dislocated and slightly rotated which caused encroachment of the vertebral canal. Whether this alteration may be responsible for the frequently observed movement disturbance of broilers in the last third of the mast period can not be decided on the basis of the pathomorphological study. In any case it must be assumed that both the pododermatitis and the bending and encroachment of the vertebral column cause pain. Thus, both lesions should be evaluated from the viewpoint of animal protection. PMID- 8721329 TI - [Ostrich farming from the perspective of veterinarians]. AB - The present paper is an attempt to relate the etiology of 54 cases with diseased/deceased ostriches to criterions of keeping, feeding, climate and infectious diseases. Despite the fact that some of the damages diagnosed seem to be avoidable the criterions of the law for the prevention of cruelty to animals are not to be fulfilled since the adverse climatic circumstances in Germany. PMID- 8721331 TI - Influence of anti-hypertensive drugs on glomerular hemodynamics. AB - Alterations in glomerular hemodynamics may play an important role in the progression of renal dysfunction. Accordingly, treating not only systemic hypertension, but also glomerular hypertension is important for conservation of renal function in patients with renal disease. However, glomerular capillary pressure does not necessarily change in parallel with systemic blood pressure due to unique mechanisms that control the resistance of glomerular afferent and efferent arterioles. While myogenic response and tubuloglomerular feedback play an important role in controlling afferent arteriolar resistance, angiotensin II is a major determinant of efferent arteriolar tone. Calcium antagonists block almost all mechanisms that constrict the afferent arteriole, rendering glomerular capillary pressure dependent on systemic pressure. On the other hand, angiotensin converting enzyme inhibitors dilate efferent arterioles, thereby lowering glomerular capillary pressure. Such differences should be taken into account when selecting adequate drugs for the treatment of hypertension with various degrees of renal dysfunction. PMID- 8721332 TI - Echography of the inferior vena cava for estimating fluid removal from patients undergoing hemodialysis. AB - Echography was used to perform 118 consecutive measurements of the diameters of the inferior vena cava (IVC) in 28 chronic hemodialysis patients. There was a significant correlation between the percent changes in the IVC in the expiratory phase (dIVC-E) and the percent changes in body weight (dBW) (r = 0.620, p < 0.002, n = 118). The average IVC diameter in the expiratory phase before dialysis (IVC-E) was 15.3 +/- 4.6 mm and this value decreased gradually following ultrafiltration and reached an average diameter of 11.0 +/- 4.3 mm after dialysis. Three cases developed hypotension due to fluid removal. Their IVC-E value showed a rapid reduction below 11.0 mm in the early phase of hemodialysis, then maintained a plateau. The hypotension was attributed to hypovolemia due to excessive dehydration. Our results suggest that the IVC-E diameter in postdialysis would approach 11.0 +/- 4.3 mm, which could be interpreted as being close to the dry weight. PMID- 8721333 TI - Role of tissue inhibitors of metalloproteinase in diabetic nephropathy. AB - We evaluated the role of tissue inhibitors of metalloproteinase-1 (TIMP-1) in patients with diabetic nephropathy by comparing the serum and urine TIMP-1 levels with those of renal biopsy specimens. A total of 35 diabetic patients were divided into four groups, D0, DI, DII and DIII-IV, according to the severity of diffuse glomerular lesions using Gellman's criteria. Using serum and 24-hour urine specimens, TIMP-1 was measured by a sandwich enzyme immunoassay. Serum and urinary TIMP-1 showed significant increases in association with the progress of glomerular diffuse lesions. There was no correlation between serum TIMP-1 and serum creatinine, creatinine clearance, serum and urinary beta 2-microglobulin, urinary NAG, HbA1c, or urinary TIMP-1. There was a significant correlation between urinary TIMP-1 and urinary albumin, and was a significant correlation between urinary TIMP-1 and urinary NAG. We conclude that TIMP-1 has a potential role in the regulation of glomerular matrix accumulation in diabetic nephropathy. PMID- 8721334 TI - Hypertension in dialysis patients: a cross-sectional analysis. AB - To clarify the current status of blood pressure control in chronic dialysis patients, we investigated a large group of patients undergoing dialysis in Okinawa, Japan. A set of cross-sectional data of 18 variables, including age, antihypertensive medication, dose of dialysis, blood chemistry findings and predialysis blood pressure, were obtained in 1,243 patients (524 females, 719 males) aged 14 to 93 years who were on hemodialysis in January 1991. Systolic and diastolic blood pressures in patients taking antihypertensive medication (160.8 +/- 0.8/84.6 +/- 0.5 mmHg, mean +/- SEM) were significantly (P < 0.0001) higher than in those were not taking antihypertensive medication (141.3 +/- 0.8/76.9 +/- 0.5). Stepwise regression and univariate analysis methods were used to determine the independent predictors of blood pressure. Antihypertensive medication was positively associated with systolic and diastolic blood pressure. However, the mortality rate of the two groups during the following four years did not differ (antihypertensive-medicated group 23.1% vs. non-medicated group 20.9%). In conclusion, hypertension does not appear to be adequately controlled in our dialysis patients, despite the widespread use of antihypertensive medications. PMID- 8721335 TI - Myocardial troponin T levels in patients with diabetic nephropathy. AB - We investigated myocardial troponin T (TnT) level as a marker for myocardial injury at various stages of diabetic nephropathy, including end-stage renal failure. One hundred and four diabetic patients were included in this study. These patients were divided into 5 groups as follows: Group I, composed of 41 patients without nephropathy who served as controls; Group II, composed of 15 patients with micro-albuminuria; Group III, composed of 15 patients with macroalbuminuria; Group IV, composed of 8 patients with renal failure who were not receiving hemodialysis; and Group V, composed of 25 patients who were receiving hemodialysis for renal failure. The following markers of myocardial injury were measured in these patients: myocardial TnT, creatine kinase (CK), myoglobin (Mb), and myosin light chain-1 (MCL-1). Our results showed that as the disease state of diabetic nephropathy advanced to renal failure, myocardial TnT levels became elevated. Group V showed significantly higher myocardial TnT levels than either Group I, Group II or Group III. Group IV showed significantly higher myocardial TnT levels than either Group II or Group III. The rate of ischemic changes on electrocardiograms also tended to increase with advance to renal failure in these patients. However, there was no correlation between myocardial TnT levels and serum Cr levels, used as an index for renal function. Myocardial TnT levels had a higher specificity for cardiac muscle than other markers for myocardial injury and are not significantly influenced by renal function. Myocardial TnT may be useful as a marker of myocardial injury for patients with chronic renal failure. PMID- 8721336 TI - A patient with sarcoidosis associated with recurrent urolithiasis and tubular injury caused by calcium deposition. AB - A 38-year-old woman was hospitalized in January 1994 with renal dysfunction and hypercalcemia. Before admission, she was diagnosed as having urolithiasis, and had been treated twice with extracorporeal shock wave lithotripsy (ESWL). Ophthalmologically, she exhibited iritis and secondary glaucoma. Hypercalcemia, an extremely low titer of parathyroid hormone (PTH), and elevation of angiotensin converting enzyme (ACE) and lysozyme activity were noted. These findings suggested sarcoidosis, although the chest X-ray showed only fibrotic changes. Hypercalcemia was suspected of having been caused secondarily by sarcoidosis. Since her laboratory data also showed renal dysfunction and abnormal urinalysis, a renal biopsy was performed. The histological findings indicated a tubular and interstitial disorder without glomerular abnormality; calcium deposition, which was detected by X-ray energy dispersive analysis, was observed in the tubular cytoplasm. Administration of prednisolone alleviated the renal dysfunction and decreased the elevation of ACE activity and lysozyme level of the blood. Sarcoidosis is sometimes associated with hypercalcemia, but rarely with renal dysfunction. These findings suggested that sarcoidosis may be associated with renal dysfunction due to tubular injury caused by calcium deposition in the tubules, and that glucocorticoid therapy was effective for these disorders. PMID- 8721337 TI - [Radiology in the past, present and future]. AB - To celebrate the year of centennial of X-ray discovery, we have to consider future development of radiology in 21st century. Since WC Rontgen discovered X ray in 1895, the radiological science have been continuously expanded. Even reviewing the history of radiology in Japan, there have been several epoch-making masterpieces. We Japanese radiologists should be more proud of Japanese radiology. It is evident that radiology of the 21st century will develop more and more on the basis of computer technology. In clinical radiology, every procedure will become more rapid and non invasive, and imaging diagnoses cover not only morphology but also function of tissue and/or cell components. To foresee the future, it is essential to understand radiology in the past and the present. In this lecture, I have reviewed a history of radiology in Japan, clarified the problems at the present and emphasized the importance of computer skill. I would like to ask all of Japanese radiologists to march forward in the midstream of medical science in cooperation of Japanese society of Radiological Technology and Industries. PMID- 8721338 TI - [Decision making and informed consent in clinical medicine]. AB - Information used in clinical medicine is always uncertain and every choice has risk of unfavorable outcome. The concept of informed consent includes full access and understanding of all relevant information by the patient with autonomic participation to the decision process. In this paper, possible cause of the lack of communication between physicians and patients is discussed along with the usefulness of decision science technology in clinical medicine. PMID- 8721339 TI - [Imaging diagnosis in border-less age]. AB - Several proposals were made under the title of imaging diagnosis of border-less age. One was the wide range of application of multimedia in medicine. The application is in the world wide scale, such as information super highway, and information technology vision 2000, and so forth. Second and third proposals were to promote clinical and basic research groups beyond specialities to make micro and micron imaging possible both in morphology and function. As an example, imaging of interstitium of the bone marrow by magnetic resonance microimaging was discussed. To convey these concepts, multidiscipline and inter- and intra speciality work is mandatory. Finally, PACS and telecommunication were introduced as one of the tools of imaging diagnosis in border-less age with illustration of PACS system at Saitama medical school. PMID- 8721340 TI - [An opinion regarding diagnostic radiology in new century]. AB - I. Technology of diagnostic radiology of the chest on next century I received and discussed concerning following subjects: 1) Dose analogue images remain on next century? 2) Progress and quality of digital images. 3) Expectation of progress of CT, MR and RI on the chest. 4) Application of monochromatic X-ray for the diagnostic imaging. For the radiologists, it is very important to assess the clinical usefulness of each diagnostic modality and form consensus development annually. II. Radiologist in the next century In accordance with the progress of technology of diagnostic radiology, it is very important to know what radiologists should do in the next century. Many referring physicians may perform not only the X-ray examinations but also therapy by themselves to ensure quality and save time from the view point of patient's benefit. Besides diagnostic imaging technologies are very attractive to them. From this point of view, radiologist should see patients and grasp the clinical data of the patients and, of course, interpret diagnostic images which were ordered by themselves. And if possible, they should treat and take care the patients. To survive in the next century, radiologist must focus on an clinical relevance and subspecialization, either by organ group or by technology. In my opinion, organ group specialization seems more useful than subspecialization according to technologies. It is considered to be a survival tactics in next century that we radiologists should band together. PMID- 8721341 TI - [Imaging science and technology in diagnostic radiology: expectations in the second century of Roentgen's discovery of X-rays]. AB - During the first century since the discovery of X-rays by Roentgen in 1885, imaging technology has contributed significantly to the progress of diagnostic radiology, mainly by providing various methods and techniques for production of diagnostic images. In the next century, it is expected that imaging science and technology will contribute to the most important process of diagnostic decision makings by radiologists and physicians by providing quantitative analyses of medical images using high-speed computers. The computer output may be used as a "second opinion" to assist radiologists' interpretation of images. This concept has been investigated as computer-aided diagnosis (CAD) during the last ten years in chest radiography and mammography for detection of lesions and characterization of normal and abnormal patterns. The aim of CAD is to improve the accuracy and the consistency of radiologic diagnoses. In this article, recent results are presented on the detection of lung nodules and pneumothoraces as well as quantitative analyses of interstitial infiltrates and cardiomegaly in chest radiographs. In mammography, CAD schemes are being developed for detection of clustered microcalcifications and masses. Recently, the prototype mammography intelligent workstation has been implemented in the clinical section of our department and initial clinical results from screening cases appear promising. PMID- 8721342 TI - [Digitized medical images]. AB - Digitalization of medical images implies that we are able to process images by computers, are able to make as many as exact copies of original images, and are able to transfer images without any loss of information. Clinical usefulness of image processing with special attention to three dimentional images were discussed. There are many problem to be solved in the development of PACS. It is important at least now that discussion of images for primary diagnosis and images for reference should be done separately. Utilization of digitized images for education is promising. A trial of education using a world wide web server-client system was reported. Possible trends of digitized medical images were described. PMID- 8721343 TI - [Interventional radiology: current and future]. AB - The current status and the future development of interventional radiology in Japan were reviewed and discussed. The recent development of interventional radiology has been supported by the technological advances in medical imaging modalities, catheters or other devices. Superselective catheterization and embolization of small vessels have been realized by digital subtraction angiography and microcatheters. Intravascular occlusion with detachable coils is an effective treatment for cerebral aneurysms. Results of balloon angioplasty for arterial occlusive disease have been promoted with the use of vascular stents. Stent-grafts for the treatment of aortic aneurysms are under clinical trials in the western countries. New devices such as specially designed balloons for local drug delivery, angioscopic catheters, intravascular ultrasound and atherectomy catheters are now clinically used. Micromachine technologies will also be applied for new therapeutic catheters. In the field of non-vascular interventional radiology, the use of open-magnet MRI and real-time MR fluoroscopy will be useful in lesion localization, needle tracking and monitoring of treatment such as interstitial laser irradiation, radio frequency tumor ablation or ethanol injection. Interventional radiology will be more widely accepted for the treatment of vascular and non-vascular diseases in the future. PMID- 8721344 TI - [Current consideration and future in radiotherapy]. AB - The basic principle of radiotherapy for cancer patients is that it has the possibility of a good result (curability), tolerable and promises a comfortable life thereafter. Radiotherapy is the important tool for cancer by conserving the organ and function and by helping improve the quality of life. Radiotherapy must be planed on the basis of considering the therapeutic ratio with a 3-dimensional treatment planning technique which is an important role in radiotherapy because they are essential in target localization. Radiotherapy is leading the way in cancer treatment for today and tomorrow. PMID- 8721345 TI - [Future prospects for nuclear medicine]. AB - Today nuclear medicine is expected to be the most sensitive approach to measure in vivo physiology, biochemistry and metabolism. Since in most of the pathophysiological process, the alternation of the organ function occurs much earlier than morphological changes. To achieve quantitatively this measurement improvements in instrumentation (SPECT, PET) and radiopharmaceuticals are essential. Nuclear physicians and researchers in many institutions and industries are continuing to provide new radiopharmaceuticals, instruments and methodologies. Nuclear medicine is defined as topographic physiological chemistry, not anatomical orientation, marking it possible to evaluate the biochemical and physiological abnormalities at the cellular level, so that it could guide to treat the patients more effectively and predict the response to the treatments. The future developement of nuclear medicine depends on an infrastructure of physics, mathematics and chemical biology. Another important role of nuclear medicine is the application of radiopharmaceuticals emitting beta ray for the treatment of diseases, especially malignant neoplasms. At present we have several options; thyroid cancer and thyrotoxicosis with I-131, neuroblastoma and pheochromocytoma with I-131-MIBG, and bone pain palliation in patients with bone metastases with Sr-89. The expansion of this field is crucial for nuclear medicine, and we believe this could happen more extensively than expected. PMID- 8721346 TI - [Radiation protection in medicine]. AB - Medical uses of radiation became by far the greatest man-made source of doses to the population at large. As technology and health care improved there has been an increase in the usage of radiation, measured by the frequency of procedures per caput and by the levels of individual and collective dose. From the early 1920's there has been the gradually developing pressure to eliminate unnecessary exposures and to reduce individual doses. Justification of practices and optimization of protection became the leading principles for keeping individual doses and the number of patients exposed As Low As Reasonably Achievable. However, less attention has been given to optimization in medical applications than in most other radiological fields. The range of doses associated with the same procedure is often extremely large, which implies that there is considerable scope for dose reduction. The first important factor is to increase awareness of the relative ease with which doses can be reduced. This requires familiarity with the levels of dose connected with the various procedures and with techniques of quality control. A new and potentially very strong tool is provided by dose constraints, which should be selected by appropriate professional bodies and which should be used in practice as a quantitative guide to good practice. The key elements of optimization of protection, based on the ICRP recommendations were discussed. PMID- 8721347 TI - [Polyamine transport in Escherichia coli and eukaryotic cells]. AB - The polyamine content in cells is regulated by both polyamine biosynthesis and its transport. We recently obtained and characterized three clones of polyamine transport genes (pPT104, pPT79 and pPT71) in Escherichia coli. The system encoded by pPT104 was the spermidine-preferential uptake system and that encoded by pPT79 the putrescine-specific uptake system. Furthermore, these two systems were ABC (ATP binding cassette) transporters consisting of four kinds of proteins: pPT104 clone encoded PotA, -B, -C, and -D proteins and pPT79 clone encoded PotF, -G, -H, and I proteins. PotD and -F proteins were periplasmic substrate binding proteins and PotA and -G proteins membrane associated proteins having the nucleotide binding site. PotB and -C proteins, and PotH and -I proteins were transmembrane proteins probably forming channels for spermidine and putrescine, respectively. Their amino acid sequences in the corresponding proteins were similar to each other. The functions of PotA and -D proteins in the spermidine-preferential uptake system encoded by pPT104 clone were studied in detail through a combined biochemical and genetic approach. In contrast, the putrescine transport system encoded by pPT71 consisted of one membrane protein (PotE protein) having twelve transmembrane segments, and was active in both the uptake and excretion of putrescine. The uptake was dependent on the membrane potential, and the excretion was due to the exchange reaction between putrescine and ornithine. In mouse mammary carcinoma FM3A cells, it was shown that the antizyme, which negatively regulates the amount of ornithine decarboxylase, also negatively regulates the activity of polyamine transport. PMID- 8721348 TI - [Adaptation and protection systems against chemical substances in living body]. AB - Author's work on the adaptation and protection systems against chemical substances, such as drugs, food additives, heavy metals, pesticides and environmental pollutants, in the living body during the past thirty five years was reviewed. Using the induction of heme and drug metabolizing enzymes (especially heme oxygenase) and of metallothionein in the liver of animals as a biological index, the adaptation and protection of these substances to the body were discussed. This is because that heme oxygenase is a heat shock protein which is induced by various environmental factor, and that metallothionein also has a wide variety of biological functions, and was identified as an acute phase protein in the first phase defense system against environmental stresses. The research under mRNA levels as more sensitive index will advance in the near future. PMID- 8721349 TI - [Properties of glycyrrhizin in Kampo extracts including licorice root and changes in the blood concentration of glycyrrhetic acid after oral administration of Kampo extracts]. AB - We investigated in vitro the properties of glycyrrhizin (GL), such as dissolution, absorption and resolution, using a Sho-Seiryu-To extract, a Sho Saiko-To extract, both including a licorice root, and licorice extract. The dissolution of GL differed with the pH of the solvent. The absorption (partition coefficient) of GL decreased with an increase in pH, and increased in the presence of other active constituents, such as baicalin, baicalein, and ephedrine. In the case of the Sho-Saiko-To extract, the conversion from GL to glycyrrhetic acid (GA) by beta-glucuronidase originated from E. coli occurred slowly. It was also suppressed by adding baicalin. We determined in vivo the pharmacokinetics of GA after oral administration of Kampo extracts in healthy volunteers. In each Kampo extract, the time of administration had no influence on the mean maximum blood concentration (Cmax) and the area under the blood concentration-time curve (AUC). Tmax was delayed in the case of the administration after meal (p < 0.05). PMID- 8721350 TI - [Studies on reversing effect of multidrug resistance by dipyridamole. I. modulation of epirubicin-induced effects on cell proliferation and cell cycle by dipyridamole]. AB - Dipyridamole, a nucleoside membrane transport inhibitor, enhanced the cytotoxicity of epirubicin for mouse leukemia P388 cells by a factor of 1.8-fold and that for 30-fold doxorubicin-resistant sublines of P388 cells (P388/DOX) by a factor of 6.5-fold. This interaction was shown to be truly synergistic by DNA histogram and median effect analysis. The dipyridamole enhancement of the cytotoxicity of epirubioin was a dose-dependent effect; it was greatest when cells were exposed to dipyridamole before treatment with epirubicin. In cell cycle experiments, 1-5 microM dipyridamole increased the accumulation of G2 + M phase produced by the treatment with 0.5-1 microM epirubicin. Dipyridamole, however, did not appear to alter the patterns of DNA histogram in sensitive cells. These results suggest that the increase of the accumulation of G2 + M phase in resistant cells is an important factor for the interaction between epirubicin and dipyridamole. PMID- 8721351 TI - [Studies on reversing effect of multidrug resistance by dipyridamole. II. Inhibition of epirubicin efflux from resistant cells by dipyridamole and its pharmacological effect]. AB - We have previously reported that dipyridamole increases the cytotoxicity of epirubicin and alters the cell cycle in doxorubicin-resistant (P388/DOX) cells, increasing the accumulation of G2/M phase by blocking the cell cycle. In cultured cells, dipyridamole increased dose-dependently the intracellular accumulation of epirubicin in the resistant cells. Simultaneous exposure of the resistant cells to epirubicin and 100 microM dipyridamole resulted in a 4.2-fold increase in proportion to the control level of epirubicin after 60 min. Dipyridamole inhibited the enhanced efflux of epirubicin in doxorubicin-resistant cells. However, dipyridamole had no effect on both the influx and efflux of epirubicin in doxorubicin-sensitive cells. In mice, lethal and bone marrow toxicity induced by epirubicin were potentiated by administration of high-dose of dipyridamole. In addition, in vivo results also demonstrated that dipyridamole in combination with epirubicin produced a significant reversal of the in vivo antitumor activity of epirubicin in mice bearing P388/DOX cells. These data imply the enhancement effects of dipyridamole on the efficacy and toxicity of epirubicin. PMID- 8721352 TI - [Antimicrobial and anti-inflammatory actions of tea-leaf saponin]. AB - Antimicrobial and anti-inflammatory actions of tea-leaf saponin, which was a mixture of saponin separated from leaves of Camellia sinensis var. sinensis, were investigated. Tea-leaf saponin showed relatively high antimicrobial activity against pathogenic dermal fungi and its MIC value for Microsporum audouinii was 10 microgam/ml. On the other hand, tea-leaf saponin inhibited rat paw edema induced by carrageenin in a dose dependent manner. Activation of hyaluronidase, one of the enzymes involved in inflammatory reactions, was inhibited by tea-leaf saponin. It was also found that tea-leaf saponin antagonized the action of leukotrien D4, one of the chemical mediators of inflammatory reactions. Any symptom of toxic reaction was not observed when tea-leaf saponin was administered orally to mice at a dose of 2000 mg/kg. PMID- 8721354 TI - [Transesophageal echocardiography as a early postoperative monitoring patients after cardiovascular surgery: analysis of 500 consecutive studies]. AB - In cardiac surgery significant residual lesions increase postoperative morbidity and mortality. To evaluate the usefulness of transesophageal echocardiography (TEE) as a early postoperative monitor in patients immediately after cardiovascular surgery, 500 consecutive patients were studied from April 1990 to December 1994. TEE was performed at the bedside in the intensive-care unit in all of these patients. TEE detected findings unsuspected by other perioperative tests or clinical examination in 103 (20.6%) of 500 patients. Imaging revealed unsatisfactory operative results that necessitated further surgery in 18 (3.6%) of the 500 patients. These data indicate that early postoperative TEE is useful in assessing immediate operative results and identifying patients with unsatisfactory results who are at increased risk for postoperative complications. PMID- 8721353 TI - [Studies on the bioactive constituents of Aurantii Fructus Immaturus]. AB - An ethanol extract of "Kijitsu" (Aurantii Fructus Immaturus, Citrus aurantium L.) collected in China was assessed for the antitumor activity using murine leukemia P388 in vivo, and the extract was found to be active by the antitumor bioassay in vivo and in vitro. The extract was separated into a petroleum ether-soluble fraction and an ethyl acetate-soluble fraction. Fractionation was carried out using an index of cell-growth inhibitory activity against mouse leukemia L1210 cells to isolate antitumor active substances or compounds. The active compounds were purified employing silica gel column chromatography and HPLC. The antitumor effect of the isolated active compounds was studied. Five compounds, auraptene, marmin, tangeretin, nobiretin and 5-[(6',7'-dihydroxy-3',7'-dimethyl-2 octenyl)oxy]psoralen were isolated from Citrus aurantium L. Though they are all known compounds, 5-(6',7'-dihydroxy-3',7-dimethyl-2-octenyl)oxy-psoralen from this plants was first isolated. These compounds showed a cell-growth inhibitory effect against L1210 and K562 in vitro. PMID- 8721355 TI - [Case report of David's operation for annuloaortic ectasia and aortic regurgitation in a Marfan patient]. AB - We report a 35-year-old Marfan patient who underwent David's operation for annuloaortic ectasia and aortic regurgitation. This new technique is done by excising the aneurysmal portion of the ascending aorta and sinuses of Valsalva but by leaving the aortic valve leaflets and some aortic wall attached to the left ventricular outflow tract inside the Dacron tube. Postoperative echocardiography revealed well coapted aortic valve leaflets and a very small amount of regurgitation. A rigid aortic route without sinuses of Valsalva may increase the mechanical stress on the leaflets and shorten their durability but this procedure have the advantage of so called Bentall's operation by preserving the native aortic valve. PMID- 8721356 TI - [A case of thymoma in an elderly patient associated with myasthenia gravis]. AB - A case of thymoma in an elderly patient associated with myasthenia gravis (MG) was reported. A 81-year-old female with growing dim of eye was diagnosed as MG, and treated with Mestinon. Two months later, she was detected to have an anterior mediastinal tumor on chest rentogenography and CT. She was diagnosed as MG (Osserman type IIA) with thymoma by Tensilon test and high anti-acetylcholine receptor antibody level. Extended thymothymectomy was performed and the tumor was encapsulated and not invasive (stage I, lymphocyte predominant type). Postoperative course was good and subjective symptoms were improved without medication. PMID- 8721357 TI - [Cardiac disfunction and myocardial energy metabolism caused by interleukin-2 (IL 2)]. AB - There is accumulating evidence that inflammatory cytokines are involved in the pathophysiology of cardiac dysfunction found in sepsis, myocardial infarction and acute rejection after heart transplantation. Although there are some previous reports on cytokines and myocardial depression, myocardial energy metabolism caused by cytokines have not been established yet. The purpose of the present study is to determine if the IL-2 effect on contractile function is related to impaired energy production. In isolated perfused rabbit hearts (n = 6), we measured developed pressure, ATP and phosphocreatine by 31P-NMR spectroscopy during and after a 5 minute infusion of IL-2 (200 U/ml/min). Although there was slightly increased inorganic phosphate which might be affect on myocardial contractility reduced, high energy phosphate and intracellular pH did not change by IL-2 infusion, suggesting another mechanism for myocardial depression caused by inflammatory cytokine, IL-2. PMID- 8721358 TI - [Lung volume reduction surgery (VRS) for lung diffuse emphysema]. AB - The lung volume reduction surgery is now being accepted for patients with lung diffuse emphysema. The basic concept of this procedure is based on reducing overall lung volume by multiple pneumectomy. The lungs are resected on both pathologic and non-pathologic lesions. As a result, elastic pull on the small airway may be achieved and normal underlying compressed lung can be re-expanded. Additionally, the compliance of thoracic wall is ameliorated with normalized thoracic volume. On the other hand, laser ablation of emphysematous lung with bullaectomy under video assisted thoracoscopy (VATS) has been performed recently. This procedure is based on reducing the non-functional air volume due to bullae underlying the pleural surface. Both procedures have still having been under consideration in last three years. In this report, the latest result of both methods in the USA are reported and discussed. Additionally, comparative data with lung transplantation, the other surgical strategy for lung emphysema, is also discussed. PMID- 8721359 TI - [Pulmonary artery banding using expanded polytetrafluoroethylene suture]. AB - Pulmonary artery banding (PAB) is essential for infants of complicated cardiac anomaly without pulmonary stenosis, especially those who require future Fontan type correction. In order to avoid pulmonary artery branch stenosis or uneven growth we utilize expanded polytetrafluoroethylene (E-PTFE) suture (Gore-Tex suture, CV-0) as a banding material. We performed three PABs (two newborns and one infant) with this suture. All children were doing well after the banding and no one revealed residual pulmonary hypertension or pulmonary artery distortion in the postoperative catheter study. One patient successfully underwent total cavopulmonary connection eight months after PAB. We believe E-PTFE suture is an advantageous material of PAB for newborns and young infants. PMID- 8721360 TI - [Emergency coronary artery bypass grafting (CABG) in a case of coronary occlusion secondary to long segmental dissection of left anterior descending artery after percutaneous transluminal coronary angioplasty (PTCA)]. AB - A 60-year-old man with unstable angina underwent PTCA at left anterior descending artery (LAD) #6, which had stenosis of 90%. Subsequently, a long segmental dissection formed from #6 to #7 and the patients was scheduled for CABG. On the day before the expected date of CABG, he had an angina attack and a 12 lead electrocardiogram (ECG) showed ST segment and T wave elevation in V1-3. Coronary angiograms revealed 99% stenosis in LAD #6 and immediately rePTCA was performed at the site. RePTCA improved the stenosis to 50%, but ECG showed an inverted T wave in V3,4 and emergency CABG was performed, with the saphenous vein at LAD #8 and high lateral artery. Some authors have stated that CABG is not useful for long segmental coronary dissection, but we conclude that CABG should be definitely considered in such a case, because this procedure resolves pressure gradients at before-and-after entry and the bypass grafts will be never occluded whenever anastomosis (intima-adventitia-graft) is performed. PMID- 8721361 TI - [Left ventricular rupture following aortic and mitral valve replacement in a patient with osteogenesis imperfecta: a case report]. AB - Intraoperative left ventricular rupture (type I) occurred following aortic and mitral valve replacement in 62-year-old man with osteogenesis imperfecta. Re replacement of the mitral valve and the repair of left ventricular rupture were successfully performed. However, the patient died suddenly 3 days after the operation. Retroperitoneal bleeding due to the insertion of a continuous hemofiltration catheter via the femoral vein was detected at autopsy. Seven patients who underwent double valve replacements for aortic and mitral disease with osteogenesis imperfecta were reported with poor surgical results. We report our patient and discuss the relationship between osteogenesis imperfecta and left ventricular rupture. PMID- 8721362 TI - [Clinical analysis of embolism with left atrial myxomas]. AB - We conducted a retrospective analysis examining the relationship between the tumor quality and the occurrence of systemic embolization in 16 patients who underwent surgical excision of the left atrial myxoma. Systemic embolization occured in 10 patients (63%) before surgery, in 5 of whom embolic episodes were the first manifestation of the tumor. Cerebral infarction had developed in 7 patients, TIA in 2, myocardial infarction in 2, limb ischemia in 2, and multiple infarctions in the kidney, adrenal gland, spleen and bone marrow in one. A permanent deficit remained in 6 patients (60%). Myxoma was a lobulated and gelatinous type in 9 (90%) of these patients with embolization and a solid type only in one case. When the dignosis of the left atrial myxoma with soft and fragile quality is made by echocardiography, surgical excision should be performed without delay to prevent systemic embolization. PMID- 8721363 TI - [A case of a patient with post-operative empyema due to Mycobacterium chelonae]. AB - We reported a case of a thoracic empyema due to M. chelonae (Atypical Mycobacterium, group IV) after thoracic surgery. A 49-year-old male underwent right middle lobectomy for pulmonary metastasis of esophageal cancer. Postoperative course was complicated for intractable air leakage, and several procedures were tried before successful re-thoractomy. Seven days after re thoractomy, mycobacterium was proved in pleural effusion. And later on M. chelonae was identified by DNA hybridization method. Therefore, open window thoracotomy was performed at once. M. chelonae disappeared 7 days after operation and the patient discharged on 40 postoperative days. Thoracic empyema by M. chelonae is rare, and only one case was reported in Japan so far. Present case was not combined with infectious pulmonary disease by some mycobacterium. Therefore it is most reasonable to suppose this intrathoracic infection developed through the thoracic drain. In conclusion, because of the M. chelonae toleranced for almost all anti-biotics including anti-tuberculous agents, except clarithromycin, the timing of surgical approach is important for the treatment of this infectious disease. PMID- 8721364 TI - [A case of tumor like RA thrombus]. AB - Seventy-year-old man, admitted with ileus, was pointed out RA tumor at routin abdominal ECHO. UCG, CT and MRI revealed RA tumor with a stalk attached to inter atrial septum. We resected the tumor and pathological study showed organized thrombus containing bacteria. We suspected that bacteremia due to the IVH catheter was the cause of the thrombus. PMID- 8721365 TI - [A case report on concomitant graft replacement of the total aortic root and the transverse aortic arch for type A acute aortic dissection associated with annuloaortic ectasia]. AB - A 49-year-old man with type A acute aortic dissection associated with annuloaortic ectasia (AAE) underwent the concomitant graft replacement of the total aortic root and the transverse aortic arch. This operation was performed with an aid of extracorporeal circulation, blood cardioplegia, selective cerebral perfusion and open distal anastomosis. The operative technique consisted of total aortic root replacement using a composite graft and total arch replacement using four vessel graft. This case suggests that type A acute aortic dissection associated with AAE involving aortic arch could be treated by concomitant graft replacement of the total aortic root and the transverse aortic arch. PMID- 8721366 TI - [A case of traumatic tricuspid regurgitation with bilateral pericardial laceration]. AB - A case of traumatic tricuspid regurgitation with bilateral pericardial lacerations is presented. The patient was a 68-year-old male with a chief complaint of dyspnea on exertion, who had had chest contusion in an automobile accident 17 years before. Two dimensional echocardiography demonstrated a systolic prolapse of the tricuspid anterior leaflet resulting in massive regurgitation. The right atrial v wave was 25 mmHg. Intraoperative findings were as follows: Three healed tears of 4-6 cm long were present in the both sides of the pericardium. The chordae tendineae of the anterior leaflet were ruptured. The tricuspid valve was replaced with a SJM valve prosthesis. To our knowledge, no case of combined tricuspid insufficiency and bilateral pericardial laceration resulting from blunt injury has ever been reported. PMID- 8721367 TI - [A case of isolated congenital mitral insufficiency]. AB - A 1-year and 8-month-old girl admitted to our hospital because of wheeze and dyspnea. Echocardiogram and cardiac catheterization confirmed isolated congenital mitral insufficiency with pulmonary hypertension. She was treated with reconstructive surgery consisting of suture of clefted anterior mitral leaflet and annuloplasty. Her postoperative course was uneventual and mitral regurgitation was remarkably improved. PMID- 8721368 TI - [A case of familial cardiac myxoma]. AB - We experienced a case of familial cardiac myxoma observed in a mother and her daughter. A 58-year-old woman was addmited to our hospital because of repeated cerebral embolism. Echocardiography showed a left atrial myxoma to be considered as the cause of cerebral embolism. At the operation, 3 myxomas were found in the left atrium, and were removed successfully. No recurrence has been observed for 14 years after the operation. Three years after that, her 3rd child, 31-year-old women, suffered from cerebral embolism and was also diagnosed as a left atrial myxoma. A friable myxoma was removed with the interatrial septum. She had no recurrence for 11 years after the operation. In patients with cardiac myxoma who have unusual biologic behavior, including familial myxoma, "complex" type myxoma must be suspected. We suggest that it is important to distinguish patients with "complex" type myxoma, because the recurrence rate is much higher in those than in patients with "sporadic" type myxoma. PMID- 8721369 TI - [Mitral valve repair by the sliding leaflet technique employed for a broad prolapse in the posterior leaflet: a case report]. AB - We report a case of a broad prolapse in the posterior leaflet successfully treated by a mitral valve repair by the sliding leaflet technique. The mitral valve regurgitation was caused by a rupture and elongation in the chordae supporting the middle scallop. First 3 cm of the leaflet was quadrangularly resected, then the posterior leaflet was detached from the annulus. Suture annuloplasty of that portion was performed and then the posterior leaflet was reattached to the annulus. Finally a Carpentier-Edwards annuloplasty ring was sutured in position. A postoperative study revealed no regurgitation. The sliding leaflet technique seemed to be effective to reduce tension in the isovolumic stress area caused by covering the broad gap left by the resected leaflet. When mitral valve repair by the sliding leaflet technique is performed, we recommend the use of an annuloplasty ring to decrease the stress on the suture line, to remodel the annular configuration and to prevent annular dilatation. PMID- 8721370 TI - [Surgery for WPW syndrome with concomitant CABG: a case report]. AB - A 58-year-old man underwent simultaneous surgery for WPW syndrome complicated by frequent attacks of atrial tachyarrhythmia combined with angina pectoris persisting after anterior myocardial infarction. Repeated PTCA of 3 times failed and restenosis occurred with recurrent angina particularly when atrial tachyarrhythmia took place. The preoperative ECG suggested the presence of a left free wall accessory pathway confirmed by an electro-physiolosical study. Coronary bypass grafting (LITA-LAD, SVG-4 PD) and division with cryoalation of the accessory pathway were performed using a transseptal left atrial approach. The patient is doing well now and free from both angina and tachycardia. PMID- 8721371 TI - [Case of aneurysmal bone cyst in sternal manubrium]. AB - A 27-year-old female who suffered from aneurysmal bone cyst in sternal manubrium, which is histologically confirmed, underwent sternal manubriumectomy. Tumor was totally resected. Manubrial defect was replaced with two metal plates, which joined both stumps of the second ribs and both sternal ends of clavicles, respectively, and wide Marlex mesh. Using non-absorbable sutures, two metal plates were fixed to the mesh, and the mesh was also secured to surrounding structures, to hold the normal function of thorax and shoulder joints. PMID- 8721372 TI - [Two cases of clear cell tumor of the lung]. AB - We report two cases of benign clear cell tumor of the lung, which were found as well-circumscribed lesions on chest X-ray films without any symptoms, and removed surgically. Light microscopy showed proliferation of clear cells filled with glycogen in both cases. Immunohistochemical examination performed in a case suggested that the tumor cells might be associated to a derivation from neural and/or smooth muscle cells, because of positive staining for NSE and SMA. In another case, the tumor formed a necrotic cavity, which could be detected by the preoperative MRI examination. This report might introduce a note of caution for the pathologic and radiographic diversity of this disease. PMID- 8721373 TI - [A case of benign cystic teratoma growing in the thoracic cavity]. AB - A case with benign cystic teratoma growing in the thoracic cavity, which is rare in Japan, in a 49-year-old woman is reported. She complained a chest pain suddenly. Chest X-ray examination revealed an abnormal shadow and pleural effusion in the right lung field. Chest CT scan and MRI disclosed a capsulated tumor in the thoracic cavity. During surgery the tumor had adhered to the part of the parietal pleura with no continuity with anterior mediastinum. The tumor measured 11.0 x 9.5 cm in size and was a mature cystic teratoma with pancreatic tissue, histologically. The fluid levels of amylase extracted from the tumor was highly elevated. Rupture due to autodigestion by pancreatic amylase of the tumor was supposed to be a cause of chest pain. PMID- 8721374 TI - [14-3-3 protein family: the structure and function in cell signaling pathway]. PMID- 8721375 TI - [Lactacystin: a tool for elucidation of proteasome functions]. PMID- 8721376 TI - [Carotenoid biosynthetic pathway elucidated at the gene level]. PMID- 8721377 TI - [Structure and function of cathepsin E, a candidate for processing proteinases]. PMID- 8721378 TI - [DNA replication licensing factor: MCM protein family]. PMID- 8721379 TI - [Mechanism of liver cell injury in type C hepatitis]. PMID- 8721380 TI - [Information from complete bacterial genome sequences]. PMID- 8721381 TI - Assessment of two clinical trials: interferon-gamma therapy in severe injury. AB - Two recent studies have examined the efficacy of interferon-gamma in reducing infection and death in patients sustaining severe injury. Both included multi center, randomized, double-blinded placebo-control design. The first trial, conducted at four university trauma centers, enrolled 213 patients, while the second trial involved nine university trauma centers and 416 subjects. Recombinant human interferon-gamma (100 micrograms) was administered subcutaneously daily for 10 days in the first trial and 21 days in the second, in addition to standard supportive therapy. In both trials infection rates were similar in the treatment arms. Although the death rate related to infection was not affected in the first study, the second trial suggested an improved outcome from this complication. The outcome of the larger trial was flawed by dominant findings at one center that had the highest enrollment, infection, and death rates. Confounding variable analysis presented here explains much of the difference between center findings in the larger trial. Thus, the benefit of interferon-gamma as an immune adjuvant in severe injury is clouded by study design flaws evaluating its use and by the inability to identify appropriate subjects using clinical criteria. PMID- 8721382 TI - The effect of methylprednisolone on myocardial beta-adrenergic receptors and cardiovascular function in shock patients. AB - The effect of methylprednisolone on the myocardial beta-adrenergic receptors after long term ( > 72 h) catecholamine infusion was studied. In 56 patients with pulmonary arterial catheter, 10 mg/kg of methylprednisolone was given as an intravenous bolus. Significant increases could be seen in cardiac output and blood pressure in patients who were simultaneously treated with vasopressors like dopamine and/or dobutamine. In patients who were on dopamine infusion higher than 10 micrograms/kg/min methylprednisolone there was an increase in the systemic vascular resistance. Patients who were not on vasopressors but received methylprednisolone, had no similar changes in hemodynamic parameters. In an in vitro analysis of tissue from the myocardium in 12/56 patients who succumbed and in four additional patients who expired after multiple trauma, a beta-adrenergic receptor assay was performed. It was found that the long term infusion of catecholamines decreased the receptor number and the methylprednisolone abolished or caused the decrease to be less pronounced. In this study we could not control the selection of the patients; a randomized study needs to be conducted in the future. PMID- 8721383 TI - Ca2(+)-dependent and Ca2(+)-independent proteinase contents in the skeletal muscle in septic rats. AB - This study assessed sepsis-induced changes in the contents of calpain and cathepsin B in rat soleus muscle. Sepsis was induced in rats by intra-abdominally implanting fecal pellets containing Escherichia coil and Bacteroides fragilis. Intact soleus muscles were isolated from non-operated control rats, and from rats sacrificed 1 and 2 days after they were implanted with bacteria-free (sterile implanted) or bacteria-laden (septic implanted) pellets. Western blot analyses of muscle homogenates were performed to identify and quantitate these proteinases using specific antibodies. No significant differences in cathepsin B contents were observed between the septic and nonseptic animals on days 1 and 2, post implantation. Among the three distinct bands recognized by anti-calpain, two prominent bands of 80 and 76 kDa, representing calpain subunits, did not seem to be altered in septic rats compared to the nonseptic groups. The content of the 45 kDa subunit decreased in both the septic and sterile groups compared with non operated control. These results along with our previous observations suggest that although Gram-negative sepsis does not appear to have an effect on Ca2(+) insensitive lysosomal cathepsin B content or activity, it upregulates the activity of the Ca2(+)-dependent calpain but not its content in the skeletal muscle during sepsis. PMID- 8721384 TI - Endotoxin interacts with tumor necrosis factor-alpha to induce vasodilation of isolated rat skeletal muscle arterioles. AB - Sepsis is characterized by decreased peripheral vascular resistance, however, discrepancies exist regarding the specific secondary mediators involved. This study examined whether the presence of endotoxin (ET) is a requirement for tumor necrosis factor-alpha (TNF-alpha) to induce vasodilation of isolated skeletal muscle arterioles. First order cremasteric arterioles were isolated from male Sprague-Dawley rats, cannulated with glass micropipettes, superfused in physiologic saline, and allowed to achieve spontaneous basal tone in the absence of intraluminal flow. A 2 min exposure to TNF-alpha (.01-100 ng/mL) had no apparent effect on arteriolar diameter (95 +/- 5% after .01 ng/mL and 92 +/- 6% after 100 ng/mL, p > .05 compared with basal). However, arterioles superfused with 2.5 micrograms/mL Salmonella enteritidis ET for 1 h followed by a 2 min exposure to 100 ng/mL TNF-alpha demonstrated a dilation (to 128 +/- 12%) that became statistically significant 10 min after TNF-alpha washout (to 142 +/- 12%, p < .05). This effect was eliminated by combined inhibition of cycloxygenase (with indomethacin) and nitric oxide synthase (L-NAME). The data indicate that neither ET or TNF-alpha alone elicit a direct vasomotor effect on the isolated arteriole preparation used in these studies. However, pretreatment of the vessels with ET results in the ability of TNF-alpha to cause arteriolar dilation, possibly through a mechanism involving both cyclooxygenase and nitric oxide synthase. PMID- 8721385 TI - Protective effects of nicotinamide against nitric oxide-mediated delayed vascular failure in endotoxic shock: potential involvement of polyADP ribosyl synthetase. AB - Nitric oxide (NO) produced by the inducible isoform of nitric oxide synthase contributes to the hypotension and vascular hyporeactivity in shock. Nicotinamide is protective against the cytotoxic effects of exogenous and endogenous NO in vitro. We investigated the effect of nicotinamide on the cellular energetic and vascular failure in a rat model of endotoxin shock. Administration of nicotinamide to rats, starting at 1 h bacterial lipopolysaccharide, maintained higher blood pressure levels, without affecting induction of nitric oxide synthase. Nicotinamide treatment prevented the lipopolysaccharide-induced decrease in mitochondrial respiration and intracellular NAD+ levels in peritoneal macrophages and improved the contractility of the thoracic aorta ex vivo. Thus, nicotinamide protects against the delayed, NO-mediated vascular failure in endotoxic shock. Its actions are unrelated to inhibition of NO biosynthesis but may be related to inhibition of the NO-mediated activation of an energy-consuming DNA repair cycle triggered by polyADP ribose synthetase. PMID- 8721386 TI - Attenuation of endotoxin-induced pathophysiology by a new potent PAF receptor antagonist. AB - The role of platelet-activating factor (PAF) as a mediator of endotoxin-induced pathophysiology has been studied in several animal models with conflicting results. We evaluated the effect of a new, potent, and specific PAF receptor antagonist, ABT-299 (Abbott Laboratories) against endotoxin (lipopolysaccharide; LPS)-induced cardiopulmonary dysfunction in a porcine model. In initial experiments, the potency of ABT-299 was confirmed in vitro by its ability to inhibit PAF-induced porcine platelet aggregation at an IC50 of .047 +/- .01 microM, and in vivo by the ability of low doses (.12 mg/kg + .03 mg/kg/h) to block the cardiopulmonary pathologic response to exogenous PAF infusion. To evaluate the effect of ABT-299 administration during endotoxemia, pigs were randomly assigned to one of three groups: controls (n = 7), LPS (n = 9), or ABT 299 + LPS (n =7). ABT-299 was given at 1.0 mg/kg from -0.5 to 0 h plus .3 mg/kg/h from 0 to 6 h. LPS was given at .5 micrograms/kg/hr from 0 to 6 h. ABT-299 reduced the early LPS-induced fall in cardiac index and stroke volume, pulmonary hypertension and vasoconstriction, bronchoconstriction, and hypoxemia. Administration of LPS resulted in 44% mortality (before 6 h), which was blocked by ABT-299. Results with this antagonist indicate that PAF contributes to endotoxin-induced cardiopulmonary dysfunction in the pig, and is associated with mortality in this model. PMID- 8721388 TI - No evidence for a protective effect of ascorbic acid on free radical generation and liver injury after hemorrhagic shock in rats. AB - Oxygen free radicals have been shown to be implicated in ischemic tissue injury, and free radical-induced reactions may also play an important role in the pathophysiology of circulatory shock. The present study was designed to investigate the potential use of ascorbic acid as an exogenous antioxidant on the liver's recovery from hemorrhagic shock in situ. Rats (fasted overnight) were subjected to 60 min of hemorrhagic shock (HS) (mean arterial pressure = 40 mmHg) under pentobarbital anesthesia, followed by retransfusion of the shed blood. One half of the animals (n = 6) were injected with 10 mg/kg of ascorbic acid prior to induction of shock, while untreated animals (n = 6) received the same volume of saline solution. in untreated animals, systemic plasma levels of malondialdehyde rose from 1.07 +/- .08 during normotension (NT) to 1.36 +/- .18* 60 min after resuscitation (RS), documenting oxygen free radical-induced lipid peroxidation. Accordingly, plasma levels of alanine aminotransferase (16.5 +/- 2.5; 34.9 +/- 12.3*; 105.8 +/- 68.7* U/L; NT/HS/RS) and ammonia (127 +/- 40; 532 +/- 160*; 304 +/- 244* micrograms/dL) rose significantly during the experiment. Hepatic ATP content of the liver fell from 4.8 +/- .83 to .56 +/- .27* after HS and recovered partially to 2.7 +/- 1.6* mumol/g after RS. Leukocyte infiltration in the liver, indicated by tissue levels of myeloperoxidase, remained constant during HS but rose during RS (37.9 +/- 18.5; 38.6 +/- 16.4; 81.4 +/- 30.7*, arbitrary units), thus documenting an inflammatory reaction after HS. In the ascorbic acid group, plasma levels of malondialdehyde were comparable to those of untreated animals after RS, as were enzyme concentrations and ammonia. No differences were observed with regard to the tissue concentrations of ATP or myeloperoxidase. Mean arterial blood pressure as well as liver tissue perfusion, as measured by Laser Doppler flowmetry, did not show significant differences between the groups. It was concluded that, although an effect of oxygen free radicals on liver tissue could be found during and after HS, treatment with ascorbic acid alone, in our model, failed to ameliorate the recovery of the animals upon resuscitation (values are mean +/- SD; *, p < .05 vs. NT; one-way ANOVA). PMID- 8721387 TI - Comparison of the capacity of rhTNF-alpha and Escherichia coli to induce procoagulant activity by baboon mononuclear cells in vivo and in vitro. AB - The procoagulant activity of mononuclear cells (MNCs) may play an important role in the disseminated intravascular coagulation seen in septic shock. This study compares the capacity of Escherichia coli (E. coli) and recombinant human TNF alpha (rhTNF-alpha) to induce procoagulant activity by baboon MNCs. In vivo studies showed that MNC procoagulant activity was significantly increased at T + 120 min after LD100 E. coli infusion into baboons. Most of this procoagulant activity was attributable to tissue factor. In contrast, a bolus infusion of rhTNF-alpha (150 micrograms/kg) and a monoclonal antibody to activated protein C (2 mg/kg) did not induce any increase of MNC procoagulant activity at T + 120 min even though the plasma TNF-alpha level was 10 times higher than that seen following infusion of E. coli. In vitro studies showed that E. coli at concentrations comparable to that observed in the vivo study and LPS at a concentration of 2.5 ng/mL induced more intense tissue factor expression by both human and baboon monocytes than rhTNF-alpha in the concentrations ranging from 10 to 1,000 ng/mL. These results suggest that TNF-alpha alone is not sufficient to induced noticeable MNC procoagulant activity, at least, in the early stage of this septic shock model. PMID- 8721390 TI - The renal effects of 7.5% NaCl-6% dextran-70 versus lactated Ringer's resuscitation of hemorrhage in dehydrated sheep. AB - A small volume of 7.5% NaCl/6% Dextran-70 (HSD) can rapidly expand the plasma volume, but concerns exist regarding its adverse effects on renal function in the dehydrated state. Sheep were thirsted for 4 days (13% plasma volume contraction), and subjected to a fixed-pressure shock model (mean arterial pressure of 50 mmHg for 2 h), followed by resuscitation with either HSD (4 mL/kg) or lactated Ringer's solution (LR; 37 mL/kg). Mean arterial pressure was restored to 90%, cardiac output to 125% and 120%, and plasma volume to 78% and 72% of baseline in LR and HSD groups, respectively. Glomerular filtration rate improved to 100% of baseline following HSD compared with 82% following LR. No significant urinary 70,000 molecular weight dextran was observed, suggesting an intact renal glomerular membrane. These data suggest that small volume HSD resuscitation is effective, even with pre-existing dehydration. In addition, renal function is not compromised by HSD resuscitation of hemorrhaged, dehydrated animals. PMID- 8721389 TI - Dibutyryl cAMP improves systemic vasoconstriction caused by endotoxin in dogs. AB - We studied whether dibutyryl cyclic adenosine monophosphate (DbcAMP), which freely penetrates into the cells, improves systemic vasoconstriction caused by endotoxin in dogs. Thirteen anesthetized dogs were randomized into three groups. The endotoxin (ETX) group (n = 5) received only Escherichia coli endotoxin (3 mg.kg-1, intravenously). The ETX + DbcAMP group (n =5) received DbcAMP (6 mg.kg 1, intravenously) 30 min before the administration of endotoxin. The DbcAMP group received the same dose of DbcAMP 30 min after administration of saline. In the ETX group, systemic blood pressure and cardiac index significantly decreased, and systemic vascular resistance significantly increased, while in the ETX + DbcAMP group, increases in systemic and pulmonary vascular resistances after the administration of endotoxin were attenuated. DbcAMP did not cause hemodynamic changes in normal dogs. Plasma concentrations in thromboxane B2 in the ETX group were higher than in the ETX + DbcAMP group. Also, the change in plasma cyclic AMP concentrations showed a good logarithmic correlation with the change in plasma thromboxane B2 concentrations after the administration of endotoxin (r = .908, log (delta T x B2) = -.002* (delta cAMP) + 3.786). We conclude that DbcAMP improves systemic vasoconstriction caused by endotoxin in dogs. The beneficial mechanism of DbcAMP on systemic vasoconstriction after the administration of endotoxin may be partially due to inhibition of thromboxane B2. PMID- 8721391 TI - Resuscitation with lactated Ringer's solution after hemorrhage: lack of cardiac toxicity. AB - The toxicity of D-lactate has been recognized for almost 30 years. This compound is found in the racemic mixture of lactated Ringer's solutions routinely used for peritoneal dialysis and the resuscitation of trauma victims. The current study was designed to investigate whether toxicity occurred at the D-lactate concentrations achieved during hemorrhage resuscitation with racemic lactated Ringer's solution. Conscious unrestrained male Sprague-Dawley rats (n = 24) were monitored for electrocardiographic abnormalities while undergoing hemorrhage and subsequent resuscitation with either L-lactated, D-lactated, or racemic lactated Ringer's solution. The rats infused with D-lactate showed significant toxicity as evidenced by bradycardia, premature ventricular contractions, and ventricular fibrillation. No such alterations were observed in the animals resuscitated with L-lactate or racemic solutions. Resuscitation with the racemic lactate mixture increased the D-lactate concentrations in the blood, but was not associated with overt changes in cardiac rhythm. The infusion of the different resuscitation fluids produced few significant differences in acid-base status of hemorrhaged rats. These findings indicate that although toxicity may be achieved with a Ringer's solution containing only D-lactate, resuscitation using the racemic mixture does not achieve D-lactate concentrations high enough to be detrimental to the animal. PMID- 8721393 TI - Determinants of prognosis in 300 episodes of infective endocarditis. AB - We studied 300 episodes of infective endocarditis in 287 patients to evaluate clinical and laboratory determinants of prognosis by estimating the probability of in-hospital death. The patients' ages ranged from 2 months to 78 (mean 30.76 +/- 16.06) years; 185 (62%) episodes occurred in male and 115 (38%) in female patients. A total of 386 complications occurred in 223 (74%) episodes of endocarditis. The infecting microorganisms were streptococci in 147 episodes, Staphylococcus aureus in 59, Staphylococcus epidermidis in 14, gram-negative bacteria in 16, other gram-positive bacteria in 8, fungi in 4. The causative microorganism was not identified in 52 episodes (negative blood cultures). The underlying cardiac disease was valvular in 119 episodes, congenital in 37, prosthetic heart valve in 69, and others in 6. No previous heart disease was identified in 69 episodes. Surgical treatment was carried out in 102 (34%) patients. Overall, 78 (26%) patients died. The probability of death was estimated with a logistic regression model (stepwise procedure). The model with best prediction included the cardiac status previous to the endocarditis, the causative microorganism, the occurrence of complications, and the blood leukocyte count. The most important variable in predicting in-hospital death was the occurrence of complications, followed by cardiac status (prosthetic valve endocarditis), the infecting microorganism, and leukocyte count. The model underestimated the severity of the disease in patients with acute endocarditis and overestimated in patients with prosthetic valve endocarditis submitted to surgical treatment. PMID- 8721392 TI - Pulmonary surfactant function following endotoxin: effects of exogenous surfactant treatment. AB - In a porcine model of endotoxin-induced adult respiratory distress syndrome (ARDS) we tested the hypothesis that the severity of lung injury would vary with the concentration of endotoxin and that reestablishment of normal surfactant function with exogenous surfactant would vary with the severity of lung injury. The therapeutic effects of exogenous surfactant treatment on pulmonary surfactant function have varied greatly in animal models of ARDS. This has created discrepancies in the literature that may be due in part to a difference in the severity of the pulmonary lesion. Yorkshire pigs were anesthetized, placed on a ventilator, and surgically prepared for hemodynamic and lung function measurements. Pigs received either 25 (25LPS) or 50 (50LPS) micrograms/kg of Escherichia coli lipopolysaccharide (LPS) followed by exogenous surfactant (SURF, 100 mg/kg) instillation, and were randomized into five groups: Control = sham LPS (n = 4); 25LPS (n = 6); 50LPS (n = 6); 25LPS + SURF (n = 5); and 50LPS + SURF (n = 6). Treatments were followed by histological and surfactant function evaluation. Histological evaluation showed the hallmarks of ARDS. Pulmonary surfactant function assessed by surfaced tension minimum (Ymin) was significantly (P < .05) elevated in both the 25LPS (20.2 +/- 2, dyne/cm) and 50LPS (19 +/- 3, dyne/cm) groups as compared with the Control group (10 +/- 1, dyne/cm). Exogenous surfactant reduced Ymin in the 25LPS + SURF group (9 +/- 2 dyne/cm, p < .05 vs. 25LPS) but not in the 50LPS + SURF group (20 +/- 1 dyne/cm, p < .05 vs. Control and 25LPS + SURF). Surfactant treatment was more effective in reestablishing normal surfactant function in animals subjected to a low dose of endotoxin, compared with animals receiving a higher dose. PMID- 8721394 TI - Hemodynamic performance of the PRIMA Edwards stentless aortic xenograft: early results of a multicenter clinical trial. AB - Between June 1991 and July 1993, 200 PRIMA Edwards stentless aortic bioprostheses were implanted in 4 European centres. Mean age of the group was 68.5 years (S.D. 8.0). The subcoronary implantation technique was used in 161 patients, the inclusion cylinder technique in 39 patients. Valve diameters ranged from 19 mm to 29 mm. Transthoracic Doppler echocardiographic studies were performed at discharge, and after 6 months and 12 months; a complete one-year follow-up was obtained. At 12 months peak systolic gradients ranged from 35.2 mmHg (19 mm valve) to 10.9 mmHg (29 mm valve) and effective orifice area ranged from 0.8 cm2 (19 mm valve) to 2.8 cm2 (29 mm valve). 30-day mortality was 3%, 12-month mortality was 5%. Complications were thrombo-embolism in six patients (3%), haemorrhage in three patients (1.5%), endocarditis in two patients (1%), and total AV-block requiring an endocavitary pacemaker in 14 patients (7%). At one year aortic regurgitation was evident in 27% patients but only one patient showed grade III. The stentless xenograft still offers good hemodynamics at one year. Morbidity and mortality are acceptable. Evaluation of the long-term performance of this new type of xenograft will be an important issue in future. PMID- 8721395 TI - Myocardial protection during reperfusion. AB - After prolonged periods of energy depletion, myocardial cells may rapidly deteriorate during the early stage of reperfusion. It has now been clearly demonstrated that this kind of acute lethal reperfusion injury is due to specific processes elicited by cellular re-energization. The most prominent single cause of acute harm to the reoxygenated myocardial cells is myofibrillar hypercontraction. Hypercontraction is caused by a resupply of energy of the myofibrils at excessive cytosolic Ca2+ concentrations. Additionally, the ability of the cytoskeleton to withstand large mechanical forces seems to be weakened after a prolonged period of energy depletion. Intracellular acidosis during the early stage of reperfusion represents a natural mechanism of protection against acute reperfusion injury. The reperfused myocardial cell may also suffer from uncontrolled water uptake and increased sarcolemmal fragility, favoring osmotic damage of cell membranes. As yet therapeutical interventions trying to specifically interfere with these pathomechanisms of reperfusion injury have only been tested experimentally. It seems promising to evaluate their utility for myocardial protection in cardio-surgical operations. PMID- 8721396 TI - Cardiac efficiency during coronary occlusion and during reperfusion after emergency revascularization under cardioprotection. AB - Myocardial infarction in consequence of a coronary artery occlusion presents a serious problem. It is the aim of any emergency revascularization to minimize the ischemia-induced damage or to salvage reversibly injured myocardium. In experiments on 8 anesthetized pigs, myocardial protection by orthograde perfusion with a high-volume cardioplegic solution was studied under controlled conditions. The left anterior descending artery (LAD) was occluded for 60 min. Then cardiopulmonary bypass was instituted and cardioplegia induced by 8 min perfusion of Bretschneider HTK solution into the aortic root. After 15 min global ischemia, the LAD was "revascularized' and 150 min reperfusion followed. Except for the early relaxation (dP/dtmin) and mean thickening velocity in the ischemic myocardium, all variables remained essentially unchanged during LAD occlusion. During the entire reperfusion, heart rate was significantly increased compared to control: 93 +/- 23 vs. 126 +/- 20/min. Left-ventricular (LV) peak pressure was significantly decreased at the end of the reperfusion, 104 +/- 33 and 77 +/- 22 mmHg, as was dP/dtmax:2155 +/- 655 vs. 1720 +/- 895 mmHg/s. Cardiac output was insignificantly decreased at the end of reperfusion, 2.6 +/- 0.6 vs. 2.4 +/- 0.5 L/min, whereas stroke-work index exhibited a significant deterioration: 1.2 +/- 0.6 vs. 0.5 +/- 0.3 mmHg.ml/kg. LV dP/dtmin was significantly impaired after ischemia and at the end of reperfusion, -1575 +/- 385 vs. -855 +/- 310 mmHg/s, while LV end-diastolic pressure exhibited only a moderate increase: 8 +/- 5 vs. 9 +/- 3 mmHg. MVO2, in turn, remained almost constant throughout the protocol for each of two methods by which it was predicted. The results show that global work, MVO2, and external efficiency were unchanged during early and late occlusion compared to control. During the entire reperfusion the myocardium was stunned, i.e. cardiac work was decreased at maintained MVO2. Thus, external efficiency was decreased. From these results we conclude that in reperfused myocardium after cardioplegic arrest, the oxygen is only inefficiently converted to develop force. PMID- 8721397 TI - Surgical patch ostiumplasty of the left main coronary artery. AB - The prevalence of ostium stenosis of the left main coronary artery, isolated or not, varies from 0.1% to 1.9%. Could ostium angioplasty and reconstruction of the proximal left main coronary artery be a real surgical alternative for this condition? What are the risks and which are the results? In the present study we have reviewed our experience of 10 patients undergoing ostium angioplasty (mean age 55 years; male/female 6/4). Four of the operations were reoperations. There was only one hospital mortality and the incidence of postoperative morbidity was low. Only one patient (a reoperation) required blood transfusion to compensate for a minor diffuse postoperative bleeding. All hospital survivors were followed up for an average period of 7 months (range 3-56). Eight of the 9 surviving patients were in perfect clinical condition at the end of the follow-up, only one was in NYHA class II and CCS class 2. The mean LVEF at the end of the follow-up was normal and the mean ergometry capacity was 159 +/- 18W. Follow-up coronary angiography showed no tendency to restenosis in any of the cases. Ostium patch angioplasty offers several advantages over conventional coronary artery bypass grafting. Reoperation, preoperative unstable angina, or poor left-ventricular function pose no contraindications for ostium angioplasty. Presence of aortic and/or ostium calcifications, however, constitutes an absolute contraindication. Ostium patch angioplasty of the proximal left main coronary artery should be considered a surgical alternative in selected patients. PMID- 8721398 TI - Coronary artery bypass grafting after failed percutaneous angioplasty compared to direct coronary bypass grafting in patients with unstable angina. AB - With the increasing experience in percutaneous transluminal angioplasty (PTCA) for unstable angina surgical revascularization versus interventional angioplasty treatment has been controversial. From 1991 to 1993, 162 patients underwent coronary artery bypass grafting (CABG) for unstable angina. While 126 patients received primary surgery (group I), 36 had a PTCA first which was followed by emergency operation for complications (group II). In group II there were more cases of single-or double-vessel disease (p < 0.001) and none had left main stem stenosis (p < 0.0001), but there was no difference in ventricular function between the groups. The number of distal anastomoses, mean cross-clamp time, and usage of the internal mammary artery (IMA) were significantly lower in group II. The peri-operative mortality rate was comparable between groups (5.5% versus 8.3%; n.s.). Myocardial infarction occurred more frequently in patients after PTCA (8.3% versus 4.0%; p < 0.05) without increasing the rate of low output syndrome. In conclusion, emergency CABG after failed PTCA in patients with unstable angina carries an acceptable perioperative risk. Since the perioperative rate of myocardial infarction is higher and the IMA is used less frequently in this setting long-term results may be better with the direct surgical approach. PMID- 8721399 TI - Inhaled nitric oxide in patients with severe heart failure: changes in lung perfusion and ventilation detected using scintigraphy. AB - Inhaled nitric oxide (NO) is a selective pulmonary vasodilator in patients with end-stage cardiac failure. Preoperative high pulmonary vascular resistance could modify early and late results after heart transplantation generally due to right ventricular failure. Aim of this study was to assess pulmonary vascular resistance variability following inhalation of NO by using a scintigraphic method. Our preliminary results suggest that inhaled NO in patients with end stage heart failure redistributes blood away from apical regions and towards more basal and posterior segments, probably dilating blood vessels in ventilated but nonperfused zones. NO may represent a simple and reliable method to evaluate dynamic response of pulmonary vasculature. PMID- 8721400 TI - Bronchogenic cancer in the elderly: operative risk and long-term prognosis. AB - Surgery for bronchogenic cancer raises the question of benefit to elderly patients. The present study reviews a cohort of 223 patients aged 70 years and more (range: 70-84), who underwent thoracotomy for pulmonary malignancy over a 10 year period. The aim of the study was to evaluate both operative risk and 5-year survival. Medical history was negative in 29% of the patients; 26% had a history of cardio-vascular disease, and 19% had a history of malignancy in complete remission. Tumor histology was squamous-cell carcinoma for 70.4%, adenocarcinoma for 24.2%, large-cell carcinoma for 3.6%, and small-cell carcinoma for 1.3% of the patients. 48.4% of patients were in stage I, 17.2% in stage II, and 30.3% in stage III. Exploratory thoracotomy was carried out in 5.8% of patients. A resection was achieved in 210 patients (pneumonectomy in 28.5%, lobectomy in 71.5%). Operative mortality was 7.2% for the whole series, 10% after pneumonectomy and 6.6% after lobectomy. Mortality was similar below and above 75 years. Overall 5-year survival was 32.9% (45.7% for stage I, 36.3% for stage II, and 13.8% for stage III). Survival was not influenced by age, symptomatic or asymptomatic presentation, medical history, and in particular not by history of malignant disease. Although operative mortality is slightly increased when compared to younger patients, long-term results legitimize surgery for bronchogenic cancer in the elderly. PMID- 8721401 TI - Carinal reconstruction under veno-venous bypass using a percutaneous cardiopulmonary bypass system. AB - We report two patients who underwent carinal resection and reconstruction for advanced lung cancer under veno-venous bypass. The bypass system consisted of a centrifugal pump and a hollow-fiber membrane oxygenator, and was established by percutaneous insertion of cannulas through the femoral veins using the Seldinger technique. This respiratory support system under veno-venous bypass was safe and useful for carinal reconstruction, providing a good visual field and adequate oxygenation. Although other standard accepted ventilation techniques can be used for carinal resection, veno-venous bypass is an alternative adjunct in cases involving difficult carinal reconstruction. PMID- 8721402 TI - Endofinger for tactile localization of pulmonary nodules during thoracoscopic resection. AB - To localize a deeply situated small pulmonary nodule during thoracoscopic resection, we developed as instrument a so-called "endofinger" for tactile localization of the nodule, and examined its usefulness. An attempt was made in 26 patients, using endofinger palpation under thoracoscopy, to localize twenty six pulmonary nodules, each of which was less than 20 mm in diameter (mean 12.2 mm) and situated apart from the pleural surface. An endofinger comprises an extended "finger' section connected with a sleeve section. The sleeve section fits with a large contract area over the surgeon's forefinger, effectively lengthening it. The surgeon uses an endofinger in each hand and palpates a pulmonary nodule with the endofinger from two directions simultaneously. Surgeons could feel the hardness of nodules with a high degree of sensitivity. The sense of touch of the endofinger is much better than that of a simple bar, due to the large contact area of the sleeve section with the real finger. With the endofinger palpation 22 of 26 nodules were localized and their margins confirmed. All of 19 nodules larger than 10 mm in size had their locality confirmed by endofinger palpation. Four of 7 nodules smaller than 10 mm in size could not be localized, however. The endofinger is a useful instrument for tactile localization of pulmonary nodules under thoracoscopic surgery, especially for nodules larger than 10 mm. The method is simple, low-cost, time-saving, and has no complications. PMID- 8721403 TI - Thoracoscopic repair of diaphragmatic hernia. AB - A 22-year-old woman presented with left subcostal pain and nausea. A radiograph and a computed tomographic scan of the chest revealed diaphragmatic hernia. Thoracoscopic surgery was performed. The herniated organs were put back into the peritoneal cavity and the hernial hilum was closed with interrupted silk suturing. PMID- 8721404 TI - A case of bronchopleural fistula detected by monitoring plasma endothelin-1. AB - Levels of endothelin (ET)-1 peptide are transiently increased after major physical stress. While studying sequential changes in plasma ET-1 levels during various types of stress, we noticed that the level of plasma ET-1 began to rise 10 days post-operatively in one patient with lung cancer who had undergone a left lower lobectomy. 35 days postoperatively a bronchopleural fistula became clinically manifest. The case is presented and the use of plasma ET-1 as an indicator is discussed. PMID- 8721405 TI - Application of saliva for drug monitoring. An in vivo model for transmembrane transport. AB - The determination of salivary drug concentrations is one of the major domains for the application of saliva in laboratory medicine. Its usefulness, however, has been criticized for many drugs because of the variability of the saliva/plasma concentration ratio (saliva/plasma ratio). Considering saliva as a model for transmembrane transport, drugs can be divided into 4 groups. The first group is characterized by a saliva/plasma ratio less than 1.0. In extreme cases a drug with negligible transport is either not detectable in saliva or is found in very low concentrations which are hard to measure or difficult to interpret. A second group leads to saliva/plasma ratios which are approximately constant and about 1.0 under most of the conditions studied. This group is ideal for monitoring salivary drug concentrations. A third group is sufficiently transferred into saliva, but the saliva/plasma ratio varies under different conditions. The reason for this variation is that the transport is influenced by several factors including an active transport mechanism. The varying influence of these factors on the saliva/plasma ratio largely depends on the physico-chemical characteristics of the particular substance. In a fourth group very high saliva/plasma ratios are observed primarily due to the degree of ionization of weak bases. A representative of each group is presented with its saliva/plasma ratio and its physico-chemical properties: ceftazidim, ethanol, digoxin and prilocain. In all cases the salivary concentration probably reflects the intracellular concentration in target tissues. All examples confirm saliva as an ideal in vivo model for the study of transmembrane transport in the human organism. PMID- 8721406 TI - Monitoring of bone metabolism after bone marrow transplantation by measuring two different markers of bone turnover. AB - Bone marrow transplant recipients may be at increased risk of osteoporosis. In a cross-sectional study we therefore measured two biochemical markers of bone turnover, bone alkaline phosphatase and the C-terminal propeptide of type I procollagen, in 22 serum samples from 9 patients before allogeneic bone marrow transplantation and 85 serum samples from 14 patients after allogeneic bone marrow transplantation. Following allogeneic bone marrow transplantation, female (but not male) patients showed elevated serum bone alkaline phosphatase values (p < 0.05). After bone marrow transplantation both female and male patients were characterized by elevated serum concentrations of the C-terminal propeptide (p < 0.01). Both the duration of cyclosporin A therapy (p < 0.05) and the time since transplantation (p < 0.01) were independent predictors of serum bone alkaline phosphatase values, whereas the duration of cyclosporin A therapy was the only independent predictor of C-terminal propeptide serum concentrations (p < 0.01). There was a correlation between bone alkaline phosphatase serum concentrations and C-terminal propeptide values in serum (p < 0.0001). These findings indicate an accelerated bone turnover in patients following bone marrow transplantation due to the stimulation of osteoblasts by cyclosporin A. In addition, oestrogen deficiency after total body irradiation may accelerate bone mass loss in female patients. PMID- 8721407 TI - Azathioprine pharmacogenetics: the relationship between 6-thioguanine nucleotides and thiopurine methyltransferase in patients after heart and kidney transplantation. AB - The commonly used immunosuppressive regimen after solid organ transplantation consists of cyclosporine A, azathioprine and steroids. Azathioprine, which is known to carry the risk of severe myelosuppression, is catabolized in vivo by xanthine oxidase and thiopurine methyltransferase, an enzyme which exhibits a common genetic polymorphism; 11% of Caucasians are heterozygous and 0.3% are homozygous with respect to thiopurine methyltransferase deficiency. Toxicity and immunosuppressive effects have been attributed to the 6-thioguanine nucleotides generated from azathioprine. We have studied thiopurine methyltransferase activity and 6-thioguanine nucleotide concentrations in erythrocytes from 39 heart and kidney recipients. Erythrocyte thiopurine methyl-transferase was determined by a radioenzymatic assay and erythrocyte 6-thioguanine nucleotide concentration with HPLC. Thiopurine methyltransferase activity [median (range, 10th-90th percentile)] was significantly (p < 0.05) higher in patients (n = 39) receiving azathioprine [285 (218-362) vs. 262 (160-352) mU/I erythrocytes] than in healthy blood donors as controls (n = 120). When stratified according to thiopurine methyltransferase phenotype, one patient homozygous for the low allele exhibited an excessive erythrocyte 6-thioguanine nucleotide concentration (2210 pmol/0.8 x 10(9) erythrocytes). Heterozygous patients had significantly higher 6 thioguanine nucleotide concentrations median: 435 pmol/0.8 x 10(9) erythrocytes) compared with concentrations in patients homozygous for the high allele (median: 86 pmol/0.8 x 10(9) erythrocytes; p < 0.01), although the azathioprine dosage did not differ (p = 0.66). Erythrocyte thiopurine methyltransferase determination therefore identifies patients at high risk of accumulating 6-thioguanine nucleotides. The monitoring of this enzyme may contribute to the safer management of immunosuppressive therapy with azathioprine. Alternative regimens such as cyclosporin A/mycophenolate mofetil or tacrolimus should also be considered for this patient group. PMID- 8721408 TI - Aprotinin prevents the development of the trauma-induced multiple organ failure in a chronic sheep model. AB - Trauma-induced multiple organ failure in sheep was prevented by aprotinin therapy. Multiple organ failure was induced in 16 female merino sheep by initial haemorrhagic shock and intramedullary femoral nailing (day 0), and 12 hourly injections of 0.75 micrograms/kg Escherichia coli endotoxin +0.7 ml/kg zymosan activated plasma (days 1-5). In addition, the aprotinin group (n = 6) received simultaneous injections of 5 mg/kg (35 695 KIU/kg) aprotinin, whereas ten animals did not receive aprotinin and served as the control group (n = 10). Organ functions were monitored for a total of 11 days by measuring haemodynamic, cardio respiratory and biochemical quantities of blood, urine and epithelial lining fluid. During the subsequent eleven day period, aprotinin induced a significant (p < 0.05) reduction of the pathological changes (development of multiple organ failure) seen in the control group. Thus, aprotinin prevented an alteration of cardiac function (cardiac index for control/aprotinin groups at day 1: 6.5/6.2, and at day 10: 10.47/7.0 1/min x m2), an impairment of lung function (mean pulmonary arterial pressure at day 1: 2.26/1.86, and at day 10: 3.83/2.13 kPa; epithelial lining fluid/plasma ratio of albumin concentrations as a direct marker of lung capillary permeability damage at day 0: 0.18/0.16, and at day 10: 0.45/0.15), a deterioration of liver function (plasma sorbitol dehydrogenase at day 0: 7.9/7.6, and at day 10: 29.6/7.4 U/1), but not of renal function (creatinine clearance at day 1: 91.4/66.1, and at day 10: 53.1/59.2 ml/min). Urinary aprotinin excretion increased up to day 3, then decreased rapidly despite further aprotinin administration. As a non-specific marker of cell damage, plasma lactate dehydrogenase indicated an aprotinin-induced organ protection (day 0: 501/409, and at day 10: 719/329 U/1). The neutrophil count and the measured chemiluminescence of neutrophils from the blood and epithelial lining fluid showed that aprotinin reduced the in vivo neutrophil activation, the alveolar neutrophil invasion, the production of inflammatory mediators, and the production of reactive oxygen metabolites during the passage of the capillary-interstitial alveolar space by neutrophils. PMID- 8721409 TI - History of computer-assisted data processing in the medical laboratory. AB - Computer-assisted processing of medical laboratory data started in the sixties. The earliest systems, which arose in English- and German-speaking laboratories, pointed the way for the development of laboratory data processing. The significance and evolution of the fundamental components of a laboratory information system, such as the placing of the request to the laboratory, identification of patients and samples, recording of data, quality control, plausibility control and results, are presented. The subject is given a wider perspective by the inclusion of a comprehensive (chronological) literature index. PMID- 8721410 TI - Use of the dry chemistry "Reflotron" blood analyzer under outdoor-field conditions in veterinary medicine. AB - Adapting the concept of "bed-side" patient analysis, the Boehringer-Mannheim Reflotron was evaluated for its possible use in veterinary medicine under outdoor field conditions. Horse blood was analysed with the Cobas Bio analyzer, and indoor and outdoor analyses were also performed with the Reflotron. All values showed close agreement with no significant differences. Good correlation coefficients (r values around 0.9000) were also seen between all methods used. The Reflotron was operated under outdoor-field conditions by using, whenever available, an on-farm electricity source, or a gas operated generator. The data supports the proposal to adapt the Reflotron (or any other proved system) for outdoor-field, patient-side blood analysis, thus providing the veterinary clinician with values for blood quantities, and enabling an improved and more complete diagnosis. PMID- 8721411 TI - A simple and sensitive assay for determination of human anti-idiotypic anti-B72.3 antibodies, which is not affected by the presence of tumour-associated glycoprotein 72. AB - An immunoradiometric assay is described for the determination of human anti idiotypic anti-B72.3 IgG. The latter is formed in ovarian cancer patients after treatment with the murine monoclonal antibody B72.3, which is directed against the tumour-associated glycoprotein 72 (TAG-72). A gel coupled with Fc-specific anti-human IgG antibodies is used as a solid phase for the extraction of serum IgG. The anti-B72.3 IgG is then specifically detected by incubation with radiolabelled B72.3 detector antibodies. Calibration standards were prepared from serum obtained from a patient repeatedly treated with B72.3 antibodies. The concentration of anti-idiotypic anti-B72.3 antibodies was expressed as TAG-72 like arb.units/1. The assay performed with two 60-minute incubation steps is characterized by a high sensitivity (detection limit: 3 x 10(3) arb.units/1) and precision (coefficients of variation: intra-assay = 6.4% and 5.8% at 80 x 10(3) arb. units/1 and 217 x 10(3) arb.units/1, inter-assay = 8.7% and 7.1% at 91 x 10(3) arb.units/1 and 212 x 10(3) arb.units/1) and a good linearity of dilution (recovery after dilution between 99% and 107%). The assay is more specific than previously described methods; no interference was observed by TAG-72 up to 3.3 x 10(7) arb.units/1. Also, non-specific human anti-mouse antibodies did not cross react up to 34.8 mg/l. The test may be modified for detection of anti-idiotypic antibodies, which are formed after treatment with other monoclonal antibodies. PMID- 8721412 TI - Variation in plasma lipid and lipoprotein concentrations in community-acquired pneumonia a six-month prospective study. AB - The nature of changes in the lipid profile caused by an acute infection is controversial. The aims of the present study were to study the changes in plasma lipids and lipoproteins in community-acquired pneumonia, to determine whether these changes differ according to the aetiologica/agents, and finally to observe the behaviour of these lipoproteins six months later. Sixty patients, aged between 18 and 87 years, admitted during the period September 1992 and April 1993 with suspected community-acquired pneumonia, were included in the study. Fifty three of the patients completed the 15-day follow-up investigation, and 37 remained available for study for up to 6 months. On admission and at 15 and 180 days, analyses were carried out for total cholesterol, HDL cholesterol, apolipoproteins A1 and B, triacylglycerols and transaminases. Student's t test for parametric variables was used for statistical analysis, and the Mann-Whitney test for non-parametric variables. The concentrations of total cholesterol (4.2 +/- 1.0 vs 5.5 +/- 1.3 mmol/1), HDL cholesterol (0.9 +/- 0.4 vs 1.2 +/- 0.3 mmol/l), apolipoprotein A1 (0.80 +/- 0.25 vs 1.15 +/- 0.28 g/l) and apolipoprotein B (0.77 +/- 0.28 vs 0.95 +/- 0.28 g/l) showed significantly lower values during the acute infectious process. These analyte concentrations became stable after 15 days with the exception of HDL cholesterol which continued to increase until 6 months (1.2 +/- 0.3 vs 1.3 +/- 0.3 mmol/l, p < 0.01). Patients with non-viral atypical pneumonia showed, on admission, higher triacylglycerol values (1.8 +/- 0.8 vs 1.3 +/- 0.9 mmol/l, p < 0.01) and lower HDL cholesterol values (0.6 +/- 0.3 vs 1.0 +/- 0.4 mmol/l, p < 0.03). Values of aspartate aminotransferase (112 +/- 117 vs 23 +/- 11 U/l, p < 0.001), alanine aminotransferase (127 +/- 141 vs 24 +/- 16 U/l, p < 0.02) and gamma-glutamyl transferase (113 +/- 158 vs 33 +/- 25 U/l, p < 0.03) were higher in the subgroup of non-viral atypical pneumonia. In conclusion, patients with community-acquired pneumonia present a significant decline in total cholesterol, HDL cholesterol and apolipoprotein A1 and B concentrations. Lower concentrations of HDL cholesterol are maintained up 15 days. Patients with non-viral atypical pneumonia present on admission significantly higher triacylglycerol and lower HDL cholesterol values. Those with non-viral atypical pneumonia also present higher transaminase values. PMID- 8721413 TI - Effects of anticoagulants on lipoprotein(a) measurements with four commercial assays. AB - Lipoprotein(a) levels in plasma are considered an independent risk factor for atherosclerosis at different sites. Although Lp(a) measurements have recently gained interest in clinical laboratories, several problems are still unresolved. A potential source of pre-analytical variability lies in the treatment of the specimens, since it has been reported that values of several lipid quantities are lower when measured in plasma instead of serum. Lp(a) was measured in serum and in EDTA-treated, heparinised and citrated plasma from 15 healthy volunteers. Four analytical methods were used: two enzyme linked immunosorbent assays [ELISA] based on a polyclonal anti-apolipoprotein(a) antibody and a polyclonal anti apolipoprotein B antibody, respectively; and two immunonephelo-metric assays [INA] based on a N antiserum to Lp(a) and on three monoclonal antibodies adsorbed on latex particles, respectively. Our measured Lp(a) values in plasma were lower than those found in serum, in particular for EDTA-treated (anti-apolipoprotein(a) ELISA: p < 0.01, anti-apolipoprotein B ELISA: p < 0.001 and Latex enhanced INA: p < 0.001) and citrated plasma (anti-apolipoprotein(a) ELISA: p < 0.05, anti apolipoprotein B ELISA: p < 0.001 and INA: p < 0.001). Lp(a) values measured in heparinised plasma were also lower than those found in serum, but the difference was not statistically significant. PMID- 8721414 TI - Ionised and total magnesium serum concentrations in renal and hepatic diseases. AB - Ionised and total magnesium concentrations were determined in the serum of different groups of patients suffering from renal or hepatic diseases. Ionised magnesium was measured by Microlyte 6 (KONE, Espoo, Finland) and total magnesium by atomic absorption spectrometry. In renal insufficiency ionised and total magnesium concentrations were almost equally increased. In proteinuria with a normal glomerular filtration rate, "pseudohypomagnesaemia" was observed, i.e. decreased total magnesium concentration in parallel with a decreased albumin concentration with no significant change in the concentration of ionised magnesium. Hypermagnesaemia occurred in liver diseases combined with renal insufficiency, whereas "pseudohypomagnesaemia" was most often found in the absence of renal failure. Also treatment with an aldosterone antagonist was associated with a normal ionised magnesium concentration, but the total magnesium concentration was decreased; when additional magnesium was administered, the total magnesium concentration approached a normal value, while ionised magnesium slightly exceeded reference values. Only during cyclosporin treatment did both ionised and total magnesium concentrations become lowered. However, the decrease of total magnesium exceeded that of ionised magnesium due to concomitant hypoalbuminaemia with reduction of the protein-bound fraction. It is concluded that especially low total magnesium concentrations should be investigated by measurement of ionised magnesium to exclude "pseudohypomagnesaemia". PMID- 8721415 TI - The 1-exp function as an alternative model of non-linear saturable kinetics. AB - Non-linear saturation kinetics can be described through a potency function, a trigonometric function, a logarithmic function, a hyperbolic function, or an exponential function. Saturable enzyme reaction kinetics can be alternatively formulated as a 1-exp function without the limitations of a steady-state assumption (d[C]/dt = 0, where C is the enzyme-substrate complex). The time dependent substrate conversion (-d[S]/dt = V(max) [1-exp(-Ka [S])]) depends on the maximum velocity (V(max)), the association constant (Ka) and substrate concentration [S]. In contrast to the classical Michaelis-Menten equation, the 1 exp function has an explicit solution for the substrate concentration [S] in an integrated form. [S] = (1/Ka) ln[1-exp(Ka [S]o)) exp(- Ka V(max) t)] A deceleration term must be introduced to describe enzyme reaction kinetics realistically. The 1-exp function with deceleration term can also be expanded to describe the three inhibition types of enzyme reaction kinetics. PMID- 8721416 TI - Biometrological uncertainty. PMID- 8721417 TI - Effect of pathological sera upon fully automated enzymatic inulin assay. PMID- 8721418 TI - Clinical interpretation of 2nd versus 3rd generation thyrotropin assays. PMID- 8721419 TI - Recommendations for the selection and use of protocols for assignment of values to reference materials. AB - It is essential that testing of patient samples give values that are traceable to those in a recognized, authorizative reference material. In addition, samples used for laboratory proficiency testing must have values assigned from such a reference material if results are to be comparable among materials and laboratories. As a result, the assignment of values to secondary and tertiary reference materials, calibrants, controls, and proficiency samples should be performed as precisely as possible, within reasonable limits. The intent of this document is to give guidelines for assignment of values at three levels of transfer. 1) from primary to secondary reference, materials, such as international or national references; 2) from secondary to tertiary reference materials, such as manufacturers' in-house calibrants and controls; and 3) from tertiary reference materials, such as manufacturers' in-house calibrants and controls; and 3) from tertiary reference materials to working calibrants and controls. It is hoped that these guidelines will facilitate the selection and utilization of an appropriate value transfer protocol for each level of value assignment. Because of the wide variety and nature of analytes, however, the guidelines are intentionally broad and may require revision for specific analytes. PMID- 8721420 TI - Laboratory work flow analysis and introduction of a multi-functional analyser. AB - Laboratory work flow analysis was performed in order to define guidelines for improved laboratory organisation and efficiency. All activities were monitored from the moment a laboratory test was requested until the result was reported and received. Detailed information was collected on numbers of samples and tests, work-stations and sample splitting, number of staff and laboratory costs and management. From the data thus obtained requirements for optimal reorganisation could be developed. Reduction of work-stations appeared to be of primary importance. This could be achieved by replacement of seven different work stations by instruments for multi-functional analysis (Cobas Integra) in the department of routine clinical chemistry. Effects of reorganisation were evaluated by repeated work flow analysis. The multifunctionality of the analysers (photometry, turbidimetry, ion selective electrodes and fluorescence polarisation) provides opportunities for efficient work structuring, avoiding the need for sample splitting, distribution of sub-samples and performing analyses at different work stations. Manual and clerical errors could thus be reduced. Laboratory service to clinicians was improved by reduction of turnaround times to such an extent that all test results are reported within 60 minutes (stat service) even during peak hours. Laboratory costs were reduced by decreasing the number of laboratory staff and work-stations. Both clinicians and patients expressed great satisfaction with the effects of this reorganisation, for which work flow analysis appeared to be an indispensable instrument. PMID- 8721421 TI - The dynamics of inflammatory cytokines in the healing process of mouse skin wound: a preliminary study for possible wound age determination. AB - The dynamics of inflammatory cytokines such as interleukin-1 alpha (IL-1 alpha), interleukin-1 beta (IL-1 beta), interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF alpha) during the healing process of mouse skin wounds were examined using the enzyme-linked immunosorbent assay and immunostaining. The applicability of this examination for wound age estimation is discussed from the perspective of forensic pathology. After wound induction, mice were sacrificed at intervals ranging from 0 to 240 h. The levels of TNF alpha and IL-1 beta began to elevate rapidly after wounding and reached a peak at 3 h. The IL-1 alpha level reached a peak at 6 h, and IL-6 peaked at 12 h. An infiltration of numerous leukocytes, indicating acute inflammation, was observed at 3 and 6 h, and the main source of the cytokines was immunohistochemically identified as neutrophils. These results indicate that TNF alpha and IL-1 beta play an important role in the commencement of inflammation. "Rebound" of cytokine levels, i.e. a re-increase, was observed at 72 h after wounding. Histological examination of the 72-h-old wound showed migration of fibroblasts and the formation of new granulation tissues, indicating the proliferative stage of the wound healing process. These experimental findings indicate that these cytokines have a close relationship to wound remodeling as well as to inflammation. From the viewpoint of forensic pathology, it is considered that inflammatory cytokines may become one of the markers for wound age estimation, but further studies are needed, especially those involving the investigation using human wound specimens with known time intervals after injury. PMID- 8721422 TI - DNA typing from human faeces. AB - A method has been developed for the forensic analysis of faeces by DNA amplification and direct sequencing of a polymorphic segment of mitochondrial DNA. Starting from as little as 10 mg wet weight of faeces, DNA was extracted by a variety of protocols and amplified using primers specific to hypervariable region 1 of the mitochondrial control region. The resulting amplification products were sequenced in solid phase using an automated DNA sequencer. In total, mtDNA sequences were generated from the faeces of nine Caucasians and compared with sequences generated from their respective blood samples. Sequences of faeces and blood samples from the same individual were identical in every case, but a range of 1-10 nucleotide differences was observed between individuals, with an average sequence variation of approximately 4.88 per 400 bp. Of the various extraction protocols assessed in this study, greatest success rates were achieved using magnetisable beads to bind and purify the DNA. STR analysis of DNA extracted from faeces was not routinely possible. PMID- 8721423 TI - Simple and sensitive detection of phencyclidine in body fluids by gas chromatography with surface ionization detection. AB - Phencyclidine (PCP) can be detected in body fluids with very high sensitivity by gas chromatography (GC) with surface ionization detection (SID) using pethidine as internal standard. PCP was extracted with Sep-Pak C18 cartridges from whole blood and urine samples, which gave clean extracts. The calibration curve for spiked whole blood was linear in the range 1.25-20 ng/ml. The detection limit of PCP was approximately 15 pg on-column (0.75 ng/ml sample), which was much lower than by GC-nitrogen phosphorus detection. The recovery of PCP and pethidine from spiked whole blood or urine samples was above 85%. This method seems very useful for the determination of PCP in forensic and clinical toxicology. PMID- 8721424 TI - Drugs in blood samples from unconscious drug addicts after the intake of an overdose. AB - This investigation includes whole blood samples from 53 drug addicts found unconscious in the Copenhagen area with evidence of a heroin overdose. Heroin/morphine was detected in 85% of the patients and other opioids in 11%. One or more benzodiazepines, most often diazepam, were detected in 75% of the patients. A blood alcohol concentration higher than 1.00 mg/g was detected in 57% of the patients. Methadone was detected in seven patients, ketobemidone in four, amphetamine in five and cocaine in one. This investigation showed widespread multi-drug abuse and heroin/morphine alone was detected in only one patient. As indicators of heroin intake, 6-mono-acetylmorphine (MAM) and morphine were detected in this investigation. PMID- 8721426 TI - Analysis of a short tandem repeat locus on chromosome 19 (D19S253). AB - A tetranucleotide tandem repeat locus on chromosome 19 (D19S253) was analysed. PCR products were detected by denaturing polyacrylamide gels with fluorescent based technology. This study has confirmed a polymorphism with 9 alleles ranging from 209 to 241 bp with a simple repeat structure arranged from 7 to 15 repeats. Family studies confirmed mendelian inheritance of alleles. The efficiency on DNA extracted from bloodstains and cigarette butts has been evaluated. The protocol has shown sensitivity and reproducibility. PMID- 8721425 TI - Firearm fatalities and injuries from hunting accidents in Germany. AB - Accidental hunting firearm injuries and fatalities (257 cases from 1961 to 1992) were evaluated in detail. Most persons responsible for the accident were more than 40 years old and experienced in hunting, and 26% of the gunshot wounds were fatal. In 77% of cases the victim was shot by another person and in 23% the wound was self-inflicted. The firearms/ammunition used were pellets from shotguns (63%), bullets from rifles (31%), shotgun slugs (3.5%) and bullets from handguns (2.5%). In 22% of all accidents from pellets severe eyeball injuries were involved, and 38% of the wounds were caused from a distance of 5 m or less, including all self-inflicted injuries. The most frequent factors responsible for the accident were: improper handling of the firearm (37%), failure to notice the victim (24.1%), covering the victim while swinging on the game (14.8%), ricocheting projectiles (13.6%), inadequate storage of the firearm (11.7%) and mistaking the victim for game (9.3%). In some cases more than one factor contributed to the accident. Defective firearm/ammunition, as the only non-human factor, was involved in only 1.6% of accidents. Some aspects of the prevention and the forensic investigation of hunting accidents are discussed. PMID- 8721427 TI - Brainstem evoked potentials in three groups of prisoners after release from detention camp. AB - Brainstem evoked potentials (BAEPs) were determined in three groups of male prisoners of war (POWs) released from detention camps and a control group. The first group comprised 21 POWs in whom BAEPs were determined 10-60 days after release (group I). The second group comprised 24 POWs in whom BAEPs were determined 6-9 months after release (group II), and the third group comprised 22 POWs in whom BAEPs were determined 12-18 months after release (group III). The control group comprised 32 subjects. The following changes were found in relation to the control group: in group I significantly longer interpeak latencies (IPLs) P1-P3; in group II significantly longer IPLs P1-P3 and P3-P5; and in group III significantly longer IPLs P1-P3. The subjective symptomatology of the POWs and the results of a routine examination indicate subclinical functional changes of the central nervous system, reflecting the dynamics of these changes. It is suggested that the basis of these changes may be a demyelinization intrathecal process, which occurred as a result of immunological changes during prolonged and intensive post-traumatic stress syndrome. PMID- 8721429 TI - Detection and quantification of lorazepam in human hair by GC-MS/NCI in a case of traffic accident. AB - A traffic accident caused by a man who declared that he was driving under influence of drugs (Temesta), led our laboratory to develop a procedure for the detection and the quantification of lorazepam in human hair. The method involves decontamination of hair with dichloromethane, incubation in Soerensen buffer (pH 7.6) in the presence of lorazepam-d4, liquid-liquid extraction with diethylether chloroform (80:20, v/v) at pH 8.4, derivatization by silylation and detection by GC-MS/NCI. The increasing concentrations of lorazepam from the end to the roots of a 16-cm-long hair strand (i.e. 31 pg/mg, 40 pg/mg and 49 pg/mg) proved that the driver had taken the drug over a long period of time. PMID- 8721428 TI - Morphometrical analysis of hemosiderin deposits in relation to wound age. AB - A morphometrical analysis of the extent of hemosiderin deposits in 71 human skin wounds with post-infliction intervals between 2 days and 7 months was performed. Earliest positive findings were detectable in a lesion aged 3 days, and with increasing wound age an increase in the amount of hemosiderin occurred. A value of more than 20% of the microscopic field with hemosiderin deposits was found earliest 8 days after wounding and therefore the detection of considerable amounts of hemosiderin (arbitrarily defined as 20% or more of the evaluated area) indicates a minimum wound age of approximately 1 week. Since the extent of hemosiderin formation depends upon the extent of the initial hemorrhage and a "physiological" reduction in the amount of this pigment with advanced wound age, slight or absent hemosiderin deposits cannot provide information on the post infliction interval. PMID- 8721430 TI - Fatal cerebellar haemorrhage due to phenprocoumon poisoning. AB - A 32-year-old patient died of a cerebellar haemorrhage and the blood coagulation analysis before death suggested defective synthesis of vitamin K-dependent clotting factors due to vitamin K deficiency. The post-mortem toxicological examination of different tissues revealed phenprocoumon poisoning as the cause of death. The differential diagnosis of vitamin K deficiency and the toxicology of hydroxycoumarins are discussed. PMID- 8721431 TI - Fatal poisoning by MDMA (ecstasy) and MDEA: a case report. AB - The first observation of lethal recreational use of MDMA (ecstasy) and MDEA in Italy is reported, together with extensive toxicological and histopathological documentation. Findings such as disseminated intravascular coagulation, rarely reported before, are colocated in the framework of the toxic syndrome for a better definition of criteria for forensic diagnosis. PMID- 8721432 TI - High-resolution vertical PAGE: an alternative electrophoretic system with multiple forensic applications. AB - DNA profiling based on PCR technology has become a powerful tool in forensic casework, as it enables specific amplification of polymorphic VNTR loci from very small quantities or even degraded human DNA. Genetic typing of AmpFLP or STR loci may require electrophoretic separation techniques usually achieved by sequencing gels. In this report we present a non-denaturing high-resolution vertical PAGE system which can be easily used for both VNTR subgroups. The system has been evaluated for single- and multiplex use in routine casework and has been shown to be rapid, sensitivite and reproducible. PMID- 8721433 TI - Hungarian population data on the loci HLA-DQ alpha, LDLR, GYPA, HBGG, D7S8 and GC. AB - Population data studies for HLA-DQ alpha and PM loci (LDLR, GYPA, HBGG, D7S8, GC) were carried out on a Hungarian Caucasian population sample of 163 unrelated individuals. Whereas the observed PM allele frequencies were similar to those reported for Caucasians, significant differences were found for HLA-DQ alpha between the Hungarian and some Caucasian population data. All six loci meet Hardy Weinberg expectations and there is no evidence for association between any pairs of loci. PMID- 8721434 TI - HumFES/FPS and HumF13B: population genetic data from north Italy. AB - DNA extracted from 119 unrelated individuals was analysed by the polymerase chain reaction at the polymorphic microsatellite loci HumFES/FPS (n = 115 individuals) and HumF13B (n = 119 individuals). The samples were collected from Caucasians living in the area of Milano (northern Italy). After horizontal polyacrylamide electrophoresis, 8 alleles were observed for HumFES/FPS, and 5 for HumF13B. Testing for Hardy-Weinberg equilibrium showed no significant deviation. The allele frequency data were compared with a German and a Turkish population sample. PMID- 8721435 TI - Local blood flux in skin and muscle during voluntary hyperventilation in healthy controls and patients with hyperventilation syndrome. AB - Laser Doppler fluxmetry was used to analyze local blood flux in the anterior tibial muscle and the skin of the calf simultaneously during 3 min of hyperventilation. Examinations were performed in 10 controls and 10 patients with a hyperventilation syndrome in order to evaluate a possible different vascular response to the provocation test. During hyperventilation, mean muscle blood flux increased in both groups. In controls, flux augmented from 23.4 +/- 12.3 arbitrary units (AU) to 51.6 +/- 28.3 AU (p < 0.05) and in patients from 21.6 +/- 10.8 AU to 45.0 +/- 26.4 AU (p < 0.05). The changes of skin blood flux during hyperventilation were not significant. The flux response did not differ significantly in controls and patients. Using the laser Doppler technique we were able to confirm muscular vasodilatation previously reported during short-term hyperventilation. Possible mechanisms include release of vasoactive substances and/or a stimulation of the autonomic nervous system. PMID- 8721437 TI - Capillary microscopy of the nailfold in healthy subjects. AB - In a total of 100 healthy subjects (36 males, 64 females) aged between 5 and 58 years, capillary microscopy of the nailfold was performed in all the fingers of both hands. The aim of this study was to determine whether capillary abnormalities, hitherto regarded as pathological, were to be found, and if so, to establish their frequency and nature. Capillary branchings - on average 4 5/person - were seen in 78% of the subjects investigated, and proved to be most common in digits IV and V of both hands. Of the subjects investigated, 94% showed meandering, 64% tortuous loops, 25% hemorrhagic extravasations, and 19% apical dilatations of the capillaries. Only a single case of a giant loop was seen. Neither avascular fields, stasis nor reversion of blood flow were seen. Maricq's venule index was increased in 3 subjects. These findings clearly indicate that the significance of deviating capillary patterns need to be reconsidered, for branchings or hemorrhagic extravasations can certainly be found in healthy subjects. PMID- 8721436 TI - Recombinant human erythropoietin inhibits the cutaneous vasodilatation induced by acetylcholine. AB - We investigated by means of telethermography the contractile response of cutaneous vessels to recombinant erythropoietin (rHuEPO) and the effects of this hormone on the vasodilatation induced by either acetylcholine, which is endothelium-dependent, and nitroprusside, which is endothelium-independent. Experiments were carried out in 12 healthy volunteers. Graded doses of rHuEPO (25, 50, 500 U/min), acetylcholine (7.5 and 15 micrograms/min), sodium nitroprusside (3 and 10 micrograms/min), and saline solution (sodium chloride 0.9%) were infused in the dorsal pedal artery of the lower limb. rHuEPO reduced the cutaneous temperature in a dose-dependent manner compared to the saline solution, thus suggesting that the hormone causes vasoconstriction. In contrast graded doses of acetylcholine and nitroprusside provoked vasodilatation: in fact both increased the cutaneous temperature compared to controls in a dose-dependent manner. The infusion of vasoconstrictive doses of rHuEPO in association with acetylcholine (15 micrograms/min) reverted the increase in the cutaneous temperature induced by the endothelium-dependent vasodilator. In contrast rHuEPO administered in combination with nitroprusside failed to block the vasodilatation induced by the endothelium-independent vasodilator. Therefore our data suggest that rHuEPO exerts an indirect vasoconstrictive effect and that acetylcholine induced vasodilatation, which is endothelium-dependent, is blunted by the vasoconstrictive activity of rHuEPO, thus demonstrating that the hormone may impair the synthesis of endothelial nitric oxide. PMID- 8721438 TI - Effects of oral administration of purified micronized flavonoid fraction on increased microvascular permeability induced by various agents and on ischemia/reperfusion in diabetic hamsters. AB - The effects of a clinically used purified micronized flavonoid fraction (S5682) containing 90% diosmin and 10% hesperidin on increased microvascular permeability induced by histamine, bradykinin and leukotriene B4 (LTB4) were investigated by intravital microscopy in the cheek pouch preparation of diabetic hamsters. We also investigated the effects of S 5682 on macro- molecular permeability increase and leukocyte adhesion during ischemia-reperfusion using the same preparation. Diabetes was induced by an intraperitoneal injection of streptozotocin (50 mg/kg). S 5682, suspended in 10% lactose solution, or vehicle (10% lactose) was administered orally for 25 days at 20 mg/kg/day (10 mg/kg twice a day), starting 5 days after the streptozotocin injection. Fluorescein isothiocyanate-labelled dextran (molecular weight 150,000) was given intravenously, 30 min after completion of the cheek pouch preparation. The leukocytes were stained by continuous intravenous infusion of acridine orange (0.5 mg/ kg/min). Histamine (2 microMs), bradykinin (1 microM), and LTB4 (0.01 microM), applied topically for 5 min, increased the number of fluorescent vascular leakage sites in postcapillary venules. A temporary ischemia (duration: 30 min) with total circulatory arrest of the cheek pouch was obtained by clamping the neck of the everted pouch. The maximum number of leaky sites (per cm2 in the prepared area) which occurs either at 5 min after the beginning of each topical application or 10 min after the onset of reperfusion was quantified in UV light microscopy. The results from 60 animals divided into ten groups of 6 animals each are presented as means +/- SEM. In comparison with vehicle, S 5682 significantly inhibited the macromolecular permeability increasing the effect of histamine (343.8 +/- 18.5 vs. 91.0 +/- 8.2 leaks/ cm2; p > 0.001), bradykinin (347.0 +/- 14.6 vs. 110.3 +/- 8.5 leaks/cm2; p < 0.001) and LTB4 (323.0 +/- 15.5 vs. 161.3 +/- 13.8 leaks/cm2; p < 0.001). At reperfusion, after 30 min ischemia, S 5682 significantly decreased the observed macromolecular permeability (168.5 +/- 19.7 vs. 52.7 +/- 6.3 leaks/cm2; p < 0.01). Flavonoid-treated animals also tended to have a lower number of leukocytes adhering to the venular endothelium (104.8 +/- 11.0 vs. 75.8 +/- 9.7/6 mm2; p > 0.05). These results demonstrate that oral administration of S 5682 for 25 days at 20mg/kg body weight/day has a protective effect on leakage of macromolecules after application of permeability-increasing substances and during ischemia reperfusion in the cheek pouch microvasculature of diabetic hamsters. In conclusion, the present data illustrating the inhibitory effect of a clinically relevant doses of S 5682 on the inflammatory processes induced in this in vivo model of microcirculation may serve as a rational basis to explain its clinical efficacy. PMID- 8721439 TI - Effects of different mediators or cytokines and monoclonal antibodies to adhesion molecules on leukocyte adhesion in rat mesenteric venules. AB - Leukocyte adhesion (LA) to the endothelium of postcapillary venules is considered to be an important step in the inflammatory response. The recruitment of blood leukocytes into sites of inflammation involves a well-coordinated and dynamic sequence of events in which several cellular adhesion molecules (CAMs) and chemotactic cytokines play an active role. The aim of the present study was to elucidate receptor-mediated interaction in mesenteric venules of leukocyte rolling/adhesion and plasma leakage. We applied intravital microscopic techniques, with the help of an analogous video image processing system, to measure changes in the microvascular integrity. Rat monoclonal antibodies (MoAb) to different CAMs were administered before inflammatory stimuli were applied. Topical application of different doses of either lipo-polysaccharide (LPS), fMet Leu-Phe, zymosan, complement C5a, Tnf-alpha, interleukin-1beta (IL-1beta), IL2 or IL-6 resulted in a dose-dependent increase in LA. The injection of a MoAb (1 mg/kg), 15 min prior to the LPS challenge, resulted in (1) total inhibition of LA, when MoAb to rat L-section, LFA1-beta and VLA-4 were used, (2) a moderate effect with LFA-1beta and Mac-1 MoAb, and (3) only a weak influence on LA by the MoAb to rat ICAM-1 (1 mg/kg). No effects were seen with IgG1 control MoAb. LA in acute models of inflammation can be regarded as a consequence of time-dependent differential effects of CAMs, as observed through the application of different MoAb. PMID- 8721440 TI - Dependence of elevated mesenteric arteriolar tone on glucocorticoids in spontaneously hypertensive rats. AB - This study is designed to explore the effect of adrenal glucocorticoids on arteriolar behavior under steady state conditions in spontaneously hypertensive rats (SHR) compared with their normotensive controls, the Wistar Kyoto (WKY) rats. Mesenteric arterioles were observed by intravital microscopy under general anesthesia and arteriolar diameters were measured. At the end of each experiment, papaverine was topically applied to determine maximum diameter of the vessels for which arteriolar tone was computed. The adrenal contribution to the atypical arteriolar tone in SHR was explored by subjecting them to bilateral adrenalectomy with or without glucocorticoid replacement therapy. Adrenalectomy led to a reduction in the elevated blood pressure in SHR to normotensive levels, which could be restored to its hypertensive level by treatment with dexamethasone. Adrenalectomy had so significant effect on blood pressure in WKY rats, although administration of dexamethasone but not aldosterone did lead to an elevation of the blood pressure in sham-operated WKY. Arteriolar tone in SHR was set at a higher steady state level than in WKY rats. In adrenalectomized SHR, tone fell to the levels observed in WKY rats. This decrease in arteriolar tone in the adrenalectomized SHR could in turn be restored to the hypertensive levels with dexamethasone but not aldosterone. Our results suggest that the enhancement of arteriolar tone, as well as the high blood pressure in SHR, may be related to the above normal levels of adrenal glucocorticoids present in the bloodstream. PMID- 8721441 TI - Rhythmical fluctuations of the intracerebral microcirculation studied in pigs. AB - Rhythmic variations in blood flow have been observed in various vascular beds, including brain. We have characterized fluctuations of the microcirculation in different locations in the brain, and studied the response to changes in arterial carbon dioxide tension, arterial pressure, and cerebrospinal fluid pressure. Laser Doppler flowmetry was performed in 20 pentobarbital-anesthetized pigs. Flow probes were positioned on the brain surface and 3, 10, and 20 mm into the cerebral tissue. The protocol included carbon dioxide breath- ing, hemorrhagic hypotension, and infusion into the cisterna magna. Twenty-five periods of low frequency oscillations (4.5/min) were found in 10 pigs with superimposed spindle shaped rhythmic variations (0.5/min) of the amplitude in 7. There were no rhythmic changes in arterial pressure or intracranial pressure. Rhythmic activity was most often seen in the probe positioned 20 mm into the brain and was often seen in several probes at the same time. Animals with rhythmic oscillations before interventions had lower cerebral perfusion pressure and arterial pressure, lower heart rate, and higher laser Doppler signal than the others. Blood loss often initiatied oscillations. High intracranial pressure tended to abolish preexisting oscillations. Hypercapnia always abolished preexisting oscillations. Oscillations were more frequent if the cerebral perfusion pressure was in the low range of cerebral autoregulation, occurred more often in the cerebral locations with relatively high local flow, were most likely to be localized, and therefore probably caused by local metabolic or myogenic variations. PMID- 8721442 TI - Hemodynamic changes associated with strangulation obstruction in cats. AB - While changes in blood flow associated with simple intestinal obstruction have been studied extensively, little is known about blood flow changes associated with strangulation obstruction. Closed loop strangulation was produced in anesthetized cats by means of a baby pressure gasket. Intestinal blood flow was measured by transit time flowmetry. Blood pressure was measured in a carotid artery and in veins of a closed loop of small intestine. The gasket pressure was increased stepwise in 10 mm Hg increments after which the intestinal venous pressure was kept constant at 50 mm Hg for 5 h. Increasing gasket pressure was followed by a corresponding increase in venous pressure in the closed bowel loop. Blood flow in the closed loop decreased with increasing venous pressure and was closely related to the arteriovenous perfusion pressure under stepwise increase of the gasket pressure and during prolonged periods with increased venous pressure. At constant elevated venous pressure the intestinal blood flow was determined by the arterial pressure. The vascular resistance in the closed loop increased exponentially with increasing venous pressure and especially at very low blood flow. In conclusion, we have found that strangulation obstruction is associated with increased venous pressure in the closed loop which contributes to maintaining intestinal blood flow during the obstruction. PMID- 8721443 TI - T-cell-rich B-cell lymphoma presenting in skin. A clinicopathologic analysis of six cases. AB - We reviewed our experience with six T-cell-rich B-cell lymphomas (TRBL) presenting in skin. Immunohistochemical studies were performed on all biopsies. The lymphoid population consisted mainly of CD3 and/or UCHL-1 (CD45RO) positive T cells. 5 to 15% of the lymphoid cells stained for the B-cell marker L26 (CD20). Monoclonality of the B-cell component was demonstrated in all cases, utilizing either light chain restriction (5 cases) or clonal immunoglobulin heavy chain gene rearrangement by polymerase chain reaction (PCR) (2 cases). One case was confirmed to be monoclonal by both techniques. Additionally, no clonal rearrangements of the T-cell receptor gamma gene were observed. There was considerable morphological variety in these cases. In H&E stained sections, the differential diagnosis included pseudolymphoma, peripheral T-cell lymphoma, Hodgkin's disease, Lennert's lymphoma and a MALT lymphoma. A significant component of monoclonal plasma cells was present in 3 of 6 cases, suggesting a possible origin from cutaneous immunocytoma. In fact, one of our cases was a biphasic lymphoma displaying TRBL with a small focus of immunocytoma. We conclude that immunophenotypic analysis is necessary for the diagnosis of TRBL. Pathologists should be aware of this type of cutaneous B-cell lymphoma to avoid misinterpretation as a pseudolymphoma. PMID- 8721444 TI - Differences of bcl-2 protein expression between Merkel cells and Merkel cell carcinomas. AB - The bcl-2 gene, originally identified in B-cell lymphomas, encodes for proteins which may assume oncogenic functions by blocking apoptosis. Bcl-2 proteins are broadly distributed among various tissues, including epithelial ones. Within the skin, bcl-2 is strongly expressed in melanocytes, but its further distribution is yet unclear. The Merkel cells, neuroendocrine-epithelial cells of the skin, are present within the epidermis and hair follicles, mostly nerve-associated, and are believed to be postmitotic and long lived. Possibly they give rise to the malignant Merkel cell carcinomas. In the present study we investigated the bcl-2 expression on the protein level by means of immunohistochemical techniques including double confocal laser scanning microscopy, as well as on the RNA level by RT-PCR techniques, in Merkel cells, Merkel cell carcinomas, and cell lines. Merkel cells were identified by double staining for cytokeratins 20 or 8/18. We demonstrate that fetal epidermal and dermal Merkel cells are immunostained for bcl-2 protein, most of them clearly weaker than melanocytes. Adult Merkel cells also express bcl-2 protein very heterogeneously, mostly weak. In contrast, Merkel cell carcinomas are usually strongly positive for bcl-2 protein with some degree of heterogeneity. This is different from malignant melanomas in which bcl-2 expression is reduced as compared to normal melanocytes. Bcl-2 gene expression was also shown for Merkel cell carcinoma cell lines on both the mRNA and the protein level. Possibly bcl-2 protein expression is downregulated during the life span of Merkel cells, arguing that they may succumb to a certain cell turnover. The comparably high bcl-2 protein level in Merkel cell carcinomas may reflect peculiar biological and clinical characteristics. PMID- 8721445 TI - Expression of basic fibroblast growth factor in desmoplastic melanoma. AB - Basic fibroblast growth factor (bFGF) is an established growth factor for melanocytes and a potent angiogenic factor. The expression of bFGF was investigated in 23 desmoplastic melanomas. (DM) (12 males, median age 64 years, and 11 females, median age 54 years) by immunostaining of formalin-fixed, paraffin-embedded sections with high-affinity purified antibody raised against recombinant human bFGF (Scios Nova, Inc.). The tumors were characterized by level II invasion in 1 case (5%), level IV invasion in 11 cases (48%), level V invasion in 8 cases (35%), and indeterminate in 3 cases. bFGF expression was observed in 22 of 23 tumors (95%), either immune localized to tumor cell nuclei in 17 of 22 tumors (77%), or to the cytoplasm of tumor cells in 5 of 22 tumors (23%). Also in these cases, bFGF was strongly expressed in the nuclei of vascular endothelial cells. Maximal expression was noted in the peripheral blood vessels of 20 tumors (91%) versus intratumoral vessels of 13 DM (59%). In conclusion, the expression of predominantly nuclear bFGF by tumor cells in DM suggests a role in mediating the desmoplastic phenotype. In addition, the localization of bFGF to vascular endothelium, particularly at the periphery of the tumor, may be relevant to tumor angiogenesis. PMID- 8721446 TI - Small-diameter invasive melanomas: clinical and pathologic characteristics. AB - BACKGROUND: Most melanomas when first diagnosed clinically are larger than common benign melanocytic nevi ( > 6 mm). Smaller melanomas, however, may occasionally occur. Yet, very little is known about them. OBJECTIVE: To determine the incidence and delineate the clinical and histologic characteristics of small diameter invasive melanomas. METHODS: Slides of all melanomas diagnosed between 1984-1993 were reviewed and measured. All invasive lesions less than 6 mm were further scrutinized clinically and morphologically. RESULTS: Among 357 invasive melanomas, 16 (4.4%) were less than 6 mm. Most lesions lacked some of the histologic features usually seen in large melanomas. One half of them were over 0.75 mm deep. Patients with small melanomas were on average 15 years younger than those with large lesions. In females, sites on the upper extremities predominate. In females, small melanomas had clinical signs of melanoma, while in males they resembled nevi. CONCLUSION: Melanomas smaller than 6 mm are not rare. Clinicians and pathologists must learn their features in order to diagnose them properly. PMID- 8721447 TI - CD44 and melanocytic tumors: a possible role for standard CD44 in the epidermotropic spread of melanoma. AB - CD44 is a polymorphic family of cell membrane glycoproteins that mediate cell matrix and cell-cell interactions involved in the mechanisms of tumor invasion and metastasis, and are subject to differential regulation during normal and malignant cell growth. We have investigated immunohistochemically the expression of CD44S and the variant isoforms CD44v3 and CD44v6 in paraffin-embedded tissue from 5 Spitz nevi, 3 compound melanocytic nevi, 2 blue nevi, 6 primary melanomas, 15 cutaneous metastases (three epidermotropic, nine dermal and three ulcerated) and 10 lymph node metastases of melanoma. Melanocytes were extensively positive for CD44S in primary melanomas and benign melanocytic proliferations. Among 15 cases of cutaneous metastases of melanoma, the three epidermotropic metastases, as well as one of the three ulcerated ones were positive for CD44S. CD44S expression was diminished or totally absent in six of the nine dermal metastases, in two of the ulcerated metastases and in seven of the ten lymph node metastases. CD44v3 and CD44v6 melanocytic expression was absent in all the lesions studied. According to our results, selective retention of CD44S expression by melanocytes in epidermotropic metastases of melanoma seems to indicate that preservation of CD44S may contribute to the intraepidermal spread of melanoma. PMID- 8721448 TI - Immune-associated cells in basal cell carcinomas of skin. AB - Increased numbers of mast cells (MCs) and lymphocytes infiltrating in basal cell carcinomas (BCCs) have been observed. The presence of these infiltrating cells has been considered a sign of an immunologic anti-tumor response in the host, but the relationship of these two cell populations has not been examined. To elucidate this possible relationship, 30 non-ulcerated BCCs were analyzed. Frozen sections of the tumors were stained with monoclonal antibodies for Langerhans' cells, lymphocyte subsets and natural killer cells. Fluorescein isothiocynate (FITC)-avidin as well as anti-tryptase and anti-CD45RO monoclonal antibodies were used on formalin-fixed, paraffin-embedded sections for mast cell and T cell identification, respectively. B cells and natural killer cells were rarely observed in these tumors. MCs and T cells were quantified by direct enumeration and expressed as number of cells per high power field (hpf). FITC-avidin and anti tryptase antibodies were equivalent in their ability to identify MCs. MC content in BCCs ranged from 1.0 to 31 cells/hpf. The number of T cells ranged from 0 to 50 cells/hpf with helper/suppressor cell ratios of 0.2 to 10. There was no correlation between helper/suppressor ratios and mast cell numbers; however, an inverse relationship was observed between the numbers of T cells and the number of mast cells in these tumors. These studies indicate that T cells and MCs are the primary immune cell populations responding to BCCs, and that decreased numbers of T cells are associated with more aggressive tumors. PMID- 8721449 TI - Infundibulocystic basaloid neoplasm. AB - Infundibulocystic basal cell carcinoma was described in 1987 as a new type of basal cell carcinoma with follicular differentiation. A great controversy followed this description, mainly about the malignant or benign character of this neoplasm and the distinction from other tumors with follicular differentiation. We present two new cases of this neoplasm: the first one had the typical features of this entity, except for the absence of stroma; the second one showed in the deep portion basaloid nodules similar to those of a nodular basal cell carcinoma. We review the criteria adduced to establish the malignant character of this neoplasm and conclude that both the nosology and biological behavior of this neoplasm with follicular differentiation remain to be determined. PMID- 8721450 TI - Sudoriferous acrosyringeal acantholytic disease. A subset of Grover's disease. AB - Three selected cases of transient acantholytic dermatosis were studied because of their definitive correlation with sweating due to fever and/ or bed-ridden situations. Biopsy specimens were serially sectioned and acantholysis was found in the acrosyringium or traced to connect to the acrosyringium in all biopsy specimens. Carcinoembryonic antigen (CEA) and eccrine gland-specific monoclonal antibody, IKH-4, were positive in acantholytic cells. Electron microscopy revealed electron dense material filling the lumen of intraepidermal eccrine ducts. This material leaked into lateral intercellular spaces of the luminal cells, passing tight junctions. Marked edema and numerous lysosomes were reminiscent of those found when eccrine acrosyringium is formed in the embryo; this suggested that an occluded and damaged eccrine intraepidermal duct was being rebuilt via lysosomal digestion. PMID- 8721451 TI - Mid-dermal elastolysis: a pathological and ultrastructural study of five cases. AB - The aim of this study was to evaluate the presence of inflammatory phenomena and elastic fiber phagocytosis in mid-dermal elastolysis. The pathological and ultrastructural features of 5 Caucasian female patients (ranging from 26 to 40 years) with acquired diffuse asymptomatic areas of skin wrinkling have been reviewed. The clinical features of all cases were characteristic of this condition and only in one patient were erythematous urticaria-like, non pruriginous patches also observed. In 4 cases a history of prolonged sun bathing was present and in 3 cases there was a short history of oral contraception. The pathological study confirmed the typical absence of elastic fibers in the midreticular dermis. In two cases elastic fibers were still detectable in the periadnexal dermis. Hematoxylin and eosin sections showed a mild perivascular infiltrate in two cases, while in three patients histiocytes were scattered among collagen bundles. Multinucleated giant cells containing fragmented elastic fibers were detectable in one patient. Ultrastructural analysis revealed large mononuclear cells with phagocytic aspects toward elastic fibers in all cases. PMID- 8721452 TI - Sjogren's syndrome plasma cell panniculitis and hidradenitis. AB - We present a 42-year-old woman with primary Sjogren's syndrome and a polyclonal gammopathy who presented with pretibial petechiae, purpura, and tender indurated plaques. The indurated plaques revealed a lobular plasma cell panniculitis, and thus Sjogren's syndrome should be added to the short list of collagen vascular diseases that can present as plasma cell panniculitis. Her biopsies also demonstrated intense perieccrine plasma cell infiltrates that may account for Sjogren's syndrome-associated hypohidrosis. We also observed occasional vascular occlusion of vessels with an amorphous, eosinophilic material possibly related to her hypergammaglobulinemic purpura. PMID- 8721453 TI - Cutaneous lymphangiectases associated with severe photoaging and topical corticosteroid application. AB - We report an elderly patient with lymphangiectases arising within skin affected by photoaging, steroid atrophy, and inflammation. Bullous lesions appeared in several sites following a year of topical corticosteroid applications for neurodermatitis. Light and electron microscopy showed that the bullae consisted of dilated lymphatic vessels within the dermis. There was also marked nodular elastosis and attenuation of dermal collagen. We suggest that this patient's lymphangiectases resulted from abnormal structure and function of the dermis due to photoaging and steroid atrophy. PMID- 8721454 TI - Malignant peripheral nerve sheath tumor of the skin: a superficial form of this tumor. AB - A case of malignant peripheral nerve sheath tumor (MPNST) of the skin, which is a superficial form of this type of tumor, is described. Although our patient did not present the usual diagnostic criteria for MPNST, the clinicopathologic findings for both the initial and recurrent tumors were compatible with those of the ordinary type of this tumor and not with those of neurotropic melanoma or cellular schwannoma. The recurrent tumor presented progression compared to the low grade malignancy in the original tumor. Perhaps because of the superficial location of the tumor our patient showed an unexpected favorable outcome in spite of repeated recurrences. Reports indicate that, of the superficial forms, the ordinary type is still less frequently encountered than the epithelioid type. We also discuss the differentiation between MPNST of the skin and neurotropic melanoma. PMID- 8721455 TI - Cutaneous pseudosarcomatous polyp: a histological and immunohistochemical study. AB - Two unusual acquired polypoid skin lesions exhibited prominent histological atypia, but were biologically benign. Both patients were elderly females. The lesions clinically mimicked fibroepithelial polyp or nevus lipomatosus. Both had been present for about 20 years. One lesion was located on the back, the other on the posterior thigh. Each lesion exhibited dilated, hyalinized vessels in the dermis with focal fibrin deposits, myxoid stroma, and a population of bizarre, pleomorphic spindle to stellate cells, some of which were multinucleated. Occasional atypical mitoses were present. One lesion had abundant admixed fat. Immunohistochemical staining was strongly positive only for vimentin. The lesions share features with degenerating angiofibroma and vaginal pseudosarcomatous polyp. As in these lesions, the atypia is most probably reactive and degenerative. PMID- 8721456 TI - Dermatopathology standards. PMID- 8721457 TI - An antiinflammatory effect of lidocaine? PMID- 8721458 TI - Small-bowel obstruction and the effects of lidocaine, atropine and hexamethonium on inflammation and fluid losses. AB - BACKGROUND: The profuse fluid losses and morbidity of patients suffering from obstructive ileus are closely related to inflammatory changes in the obstructed bowel wall. Previous experimental studies have shown that use of steroids and NSAIDs can reduce fluid losses in obstructive ileus. In the present study, we investigated the effects of lidocaine on fluid losses since local anesthetics have been shown to possess wide and potent anti-inflammatory properties. Hexamethonium and atropine were used to study the importance of the autonomic nervous system in bowel obstruction. METHOD: Experiments were performed in rats in vivo. After 18 h of total obstruction of the jejunum by thread ligation, a segment of the obstructed jejunum was placed in a chamber with intact nervous and vascular supply and net fluid transport was continuously registered by a gravimetric technique. Extravasation of Evans blue albumin as marker of inflammation was quantified by spectrophotometry. RESULTS: Hexamethonium (10 mg.kg-1 i.v.) significantly inhibited net fluid secretion (P < .05), while atropine (0.25 mg.kg-1 i.v.) had no significant effect. Net fluid secretion was reversed into absorption following an intravenous bolus dose of lidocaine (2 mg.kg-1) (P < 0.01) and topical administration of lidocaine (20 mg) on the serosa of the obstructed gut (P < 0.01). Single topical administration of lidocaine (20 mg) immediately before ligation significantly reduced net fluid secretion (P < 0.05) and inflammation (P < 0.05) in the obstructed bowel 20 h post-ligation compared to obstructed controls. CONCLUSION: Lidocaine significantly inhibited or prevented fluid losses when administered intravenously or topically to the obstructed gut. Mechanisms of action could be inhibition of nerve reflexes involved in fluid secretion and by inhibition of inflammation in the bowel wall. The inhibition of fluid losses by hexamethonium further supports the importance of the autonomic nervous system in the pathophysiology of bowel obstruction. PMID- 8721459 TI - Extracorporeal membrane oxygenation (ECMO) as lung or heart assist. AB - Extracorporeal membrane oxygenation (ECMO) may serve as extracorporeal lung assist (ECLA) in patients with acute respiratory failure (ARF) or as extracorporeal heart assist (ECHA) in patients with low output syndrome (LOS) after open heart surgery. From 1988 to 1992 seven patients underwent ECMO in our hospital; four suffered from ARF and three from LOS. Various bypass techniques were employed. Two ARF patients, aged 58 and 18 years, had veno-venous bypass; in the latter, ECMO was reinstituted as a veno-arterial bypass one week after weaning. In a three-year-old boy, the ECMO outflow tubing was primarily connected to the pulmonary artery, and shortly afterwards relocated to the common carotid artery. In a 31-year-old man with ARF, and three LOS patients, a 56-year-old woman, and two men aged 68 and 70 years, ECMO was veno-arterial with direct access to the ascending aorta. A heparin-coated system was used, and all but one patient, who was treated with warfarin, received a daily low dose of heparin, which was withdrawn after from one to nine days. Six patients were weaned off ECMO after 4.5 to 21 days. Three ARF patients recovered completely; the child died. In one LOS patient, ECMO was withdrawn due to a poor general condition. Two others were weaned off ECMO and the intra-aortic balloon pump, and the inotropic support was significantly reduced, but both died of multiple system organ failure. Although no firm conclusions can be drawn from these few case reports, the heparin-coated system used as ECLA appears promising, whereas ECHA seems to imply a poor prognosis in patients who are not candidates for cardiac transplantation. PMID- 8721460 TI - Safety aspects of delivery and monitoring of nitric oxide during mechanical ventilation. AB - In the presence of oxygen NO is oxidised to NO2, which is toxic in higher concentrations. In this technical investigation, we evaluated a dosage system, modified from Stenqvist et al. 1993 (1), regarding NO and NO2 levels. NO was administered before the ventilator and NO2 scavenged using a soda little absorber in the inspiratory limb close to the ventilator. NO/NO2 levels were measured using fuel cell technique. We tested the duration of soda lime scavenging, put in additional soda lime absorbers, used charcoal as absorber and exchanged tubing material. NO was delivered after the ventilator and we studied effect of interruption of ventilation. With concentrations of NO at or below 40 parts per million (ppm) at F1O2 0.9, NO2 levels were 1.2 ppm or lower. Corresponding values for 20 and 10 ppm were 0.4 and 0.2 ppm, respectively. Duration of the soda lime absorber was at least 72 hours. Additional soda lime absorbers did not further reduce NO2 levels. Charcoal absorbers reduced NO2, but also NO by 45% from set value. Tubing materials had no influence on NO and NO2 levels. When administering NO at the Y-piece, levels of NO were increased by 35-60% and NO2 levels by 110 230% compared to set values. Oxidation of NO to NO2 is continuously taking place in the breathing system. Doses of up to 40 ppm NO should be considered safe regarding NO2 levels. Administration of NO at the Y-piece gives high and unpredictable levels of NO2. PMID- 8721461 TI - Heparin-coated circuits reduce the formation of TNF alpha during cardiopulmonary bypass. AB - BACKGROUND: Cardiopulmonary bypass (CPB) causes a systemic inflammatory response. TNF alpha, which is a major inflammatory mediator, has been found in the circulation during and after CPB. Although previous studies have shown that heparin coating of the extracorporeal circuits reduces complement and granulocyte activation, and the inflammatory response, the possible effect of heparin coating on TNF alpha formation and the inflammatory response has not been fully investigated. METHODS: Eighteen patients scheduled for coronary artery bypass grafting were divided randomly into two groups. One group of patients had extracorporeal perfusion using heparin coated circuits (HC group, n = 9). The other group had extra-corporeal perfusion using an identical circuit that was not coated (UC group, n = 9). Blood samples were drawn before, during, and after CPB for measurement of plasma TNF alpha, plasma IL-8, neutrophil count, and neutrophil elastase. RESULTS: Plasma levels of TNF alpha increased during CPB in the UC group but not in the HC group. Plasma concentrations of IL-8 increased similarly during and after CPB in both groups. Coating the circuits with heparin did not affect the levels of IL-8. In both groups, the neutrophil count increased after the release of the aortic cross clamp and remained elevated for three days. In the HC group, however, the increase of neutrophil count was significantly lower compared with the UC group. Plasma concentrations of neutrophil elastase were significantly increased during and after CPB in both groups. However, the levels of elastase were significantly lower at certain time points in the HC group. CONCLUSION: From these observations, we conclude that heparin coating of the extracorporeal circuits reduces the TNF alpha formation during CPB, which may reduce neutrophil activation. PMID- 8721462 TI - Lidocaine attenuates hyperoxic lung injury in rabbits. AB - BACKGROUND: High concentrations of oxygen acute lung injury. Neutrophils are thought to play a pivotal role in the pathogenesis of this lung injury through the release of oxygen radicals, neutral proteases, and lysosomal enzymes. Lidocaine has been shown to inhibit neutrophil function. We examined whether intravenous pretreatment with lidocaine attenuated acute lung injury induced by hyperoxia. METHOD: Twenty-seven anaesthetized male rabbits were allocated to receive one of three treatments (n = 9 for each group): ventilation with 100% oxygen for 36 h with and without lidocaine treatment, and ventilation with air for 36 h without lidocaine. In the lidocaine-treated group, a single intravenous lidocaine 2 mg/kg was administered immediately after the initiation of exposure to 100% oxygen; thereafter, the lidocaine was infused at a rate of 2 mg.kg(-1).h( 1) for 36 h until the animals were sacrificed. Haemodynamics, PaO2, and lung mechanics were recorded during the ventilation period. After exposure, the lung mechanics and cell fraction in bronchoalveolar lavage fluid (BALF) were measured and analyzed, as was the concentration of activated complements, and cytokines in BALF. The lung wet-to dry- (W/D) weight ratio and albumin concentrations in BALF were analyzed as an index of pulmonary oedema. We also compared the chemiluminescence of neutrophils isolated from the pulmonary artery, and light microscopic findings, in the three groups. RESULTS: Pure oxygen for 36 h caused no significant changes in haemodynamics, lung mechanics, or PaO2/FiO2 ratio. However, hyperoxia significantly increased the lung W/D weight ratio, the influx of neutrophils into the lung, and BALF concentrations of C3a, C5a, TNF-alpha, IL 1 beta, and albumin. Lidocaine attenuated these increases (W/D ratio: 5.7 vs 5.1, %PMN: 19.2% vs 1.6%, C3a: 62 ng/dl vs 14 ng/dl, C5a: 7.9 ng/dl vs 4.1 nd/dl, TNF alpha: 25 fmol/ml vs 2.8 fmol/ml, IL-1 beta: 36 fmol/ml vs 2.2 fmol/ml, and albumin: 9.5 mg/dl vs 2.8 mg/dl, all: P < 0.05). The chemiluminescence was increased in hyperoxic compared with in normoxic rabbits and lidocaine treatment attenuated the increase (opsonized zymosan stiluated: 3.7 x 10(6) cpm vs 2.3 x 10(6) cpm, P < 0.05). Exposure to 100% oxygen caused extensive morphologic lung damage which was lessened by lidocaine (lung injury score (mean): 3 vs 4, P < 0.05). CONCLUSION: These findings suggest that intravenous lidocaine has a prophylactic effect on initial hyperoxic lung injury (pulmonary vascular permeability, histopathological, and biochemical BALF changes) in rabbits. The effects of lidocaine on more severe lung injury (decreased oxygenation) caused by hyperoxia for 72 h deserve further study. PMID- 8721463 TI - Low-dose dopamine in surgical and intensive care unit patients. PMID- 8721464 TI - Patient-controlled premedication by i.v. midazolam for ambulatory surgery. AB - Applying the principle and equipment of patient-controlled analgesia, this double blind randomised study was designed to determine the premedication dose of midazolam and to provide information about the need for preoperative anxiety control. The effects of patient-controlled premedication by i.v. midazolam were compared to those of a 1-hour i.v. infusion of a fixed dose of 4 mg. Two groups of 25 patients were studied prior to ambulatory surgery. Using a visual analogue scale graded from 0 (no anxiety) to 100, anxiety and short-term memory as well as vital signs, vigilance, cognition and orientation were assessed before premedication and then every 20 min until admission to the operating room. In both groups, 40% of the patients were free of anxiety before premedication. Sixty four percent of patients in the self-pre-medicated group never used the pump. The dose of midazolam was only 0.7 0.2 mg (mean SD) in this group taken as a whole, 0.7, 1.1 mg (mean SD) in the 8 patients who had an anxiety score at least equal to 50 before premedication, and 1.7 0.3 mg (mean SD) in the 9 patients who chose to push the button of the patient-controlled device. Anxiety and short-term memory scores decreased significantly and similarly in the self-premedicated group and in the fixed-dose group. For similar intraoperative anaesthetic requirements, time from the end of anaesthesia for determining whether a patient was ready for admission to the postrecovery lounge was shorter in the self premedicated group (26 +/- 8 min vs 32 +/- 8 min; mean +/- SD. Reduced doses of midazolam self-administered via a patient-controlled device can result in a relaxed preoperative period and amnesia in ambulatory surgery patients. However, the level of anxiety before premedication was low, which calls into question the legitimacy of the patient-controlled premedication in this kind of population. The expense of the pump was not justified by the small number of patients using it, probably because they were worried by the technical nature and the invasiveness of this technique. PMID- 8721466 TI - The effect of continuous epidural infusion of ropivacaine (0.1%, 0.2% and 0.3%) on nerve conduction velocity and postural control in volunteers. AB - BACKGROUND: Continuous epidural infusions of local anaesthetics have become increasingly popular in postoperative pain treatment, especially as they permit early mobilisation. Ropivacaine is a promising new agent which induces more pronounced sensory than motor blockade. This study was focused on the influence of continuous epidural infusion of ropivacaine on impulse conduction in large nerves (by measurement of F and H latencies), and on the subjects' ability to maintain postural control during mobilisation. METHODS: Healthy male volunteers received 0.1%, 0.2% or 0.3% ropivacaine, and bupivacaine 0.25% was used as reference. A bolus epidural injection of 10 ml of the drug, at L2/3 level, was followed by continuous infusion at 10 ml/h for 21 h. Motor blockade was assessed by mechanical measurements of force during big toe flexion and by recording of F latency. Sensory blockade was monitored by pin-prick and Thermotest methods, and by H latency recording. The subjects' ability to perform a postural test was evaluated by posturography. RESULTS: The F and H latencies became prolonged/abolished dose-dependently. With ropivacaine, F latency recovered significantly later than motor function (P = 0.0002), and H latency recovered later than normal pin-prick perception (P = 0.0006). However, the duration of partial blockade of thermoperception was comparable to that of H latency prolongation. Posturographically, the subjects receiving 0.1% ropivacaine differed significantly from all others (P < 0.001) in that they were able to maintain postural control during the infusion. The recovery period after termination of infusion was significantly shorter with ropivacaine than with bupivacaine for all measured variables. CONCLUSION: Recovery of postural control with 0.2% and 0.3% ropivacaine is significantly faster than with bupivacaine 0.25%. H latency recording allows detection of epidural blockade intensity that does not prevent subjects from performing postural tests. PMID- 8721465 TI - In vitro myocardial depression by ketamine or thiopental is dependent on the underlying beta-adrenergic tone. AB - BACKGROUND: Depression of myocardial contractility by muscarinic stimulation is dependent on the underlying beta-adrenergic tone. Prior beta-adrenergic stimulation enhances muscarinic negative inotropic responses, an effect that has been termed accentuated antagonism. The purpose of this study was to determine whether accentuated antagonism occurs with myocardial depression caused by thiopental or ketamine. METHODS: Using an isolated, electrically stimulated rat left afrium model, the dose-response curves to the muscarinic agonist carbachol and the anesthetics ketamine and thiopental were compared under conditions of high (10(-6)M isoproterenol bath concentration) or low (10(-6)M propranolol) beta adrenergic tone. RESULTS: As expected, depression by carbachol was accentuated in preparations stimulated with isoproterenol compared with atria treated with propranolol. The decrease in tension by high doses (> 400 muM thiopental, > 200 muM ketamine) of thiopental or ketamine was attenuated in isoproterenol stimulated tissue when compared with beta-adrenergic blocked muscle. Low concentrations (200 muM thiopental, 100 muM ketamine) of anesthetic caused either no change in contractility (thiopental) or small positive inotropic responses (ketamine) in pro-pranolol-treated but not isoproterenol-stimulated tissue. CONCLUSIONS. In contrast to muscarinic agonists, myocardial depression by high concentrations of ketamine or thiopental is attenuated by prior beta-adrenergic stimulation. Positive inotropic responses may be seen with low concentrations of ketamine in muscle with low beta-adrenergic tone. The results of this study demonstrate that the underlying beta-adrenergic tone greatly influences the in vitro response of cardiac tissue to ketamine or thiopental. PMID- 8721467 TI - Pharmacodynamics of subarachnoid hyperbaric 5% lignocaine. AB - Twenty male patients (55 to 85 yr) undergoing cystoscopy received spinal anaesthesia with either 75 mg (n = 10) or 100 mg of hyperbaric lignocaine 5% under standardised conditions. Plantar flexion muscle power was recorded during onset and offset of anaesthesia using a load cell interfaced with a computer (PFPD) and these data were compared with intermittent clinical assessments of spinal anaesthesia. Onset of paralysis was rapid and complete with motor power declining exponentially to 5% of preoperative values by 5 minutes in all patients. Extent of block to cold and pinprick was similar in both dosage groups (median T4), as was the rate of block onset. Block regression was complete by three hours in all patients and restitution of plantar flexion motor power was associated with normal thermosensibility at L1 and recovery of the patient's ability to walk and micturate. Recovery of plantar flexion motor power occurred at 95.5 +/- 7.38 min in patients given 75 mg compared with 129 +/- 9.5 min in those given 100 mg lignocaine (P < 0.05). The time between onset and full motor recovery in the 75 mg group (7.2 +/- 1.2 min) was less than the 100 mg lignocaine group (29 +/- 5.1 min) (P < 0.001). The larger dose of lignocaine did not confer any clinical advantage in block onset or intensity and made the onset of recovery less predictable. The Bromage grading, while clinically appropriate during anaesthesia onset, does not provide data relating to the density of block and the PFPD was therefore useful for describing the anaesthesia recovery phase. PMID- 8721468 TI - Is preoperative ketorolac a useful adjunct to regional anesthesia for inguinal herniorrhaphy? AB - BACKGROUND: Nonsteroidal antiinflammatory drugs (NSAIDs) have become a popular component of analgesia regimens, particularly in combination with narcotics. We questioned whether there might also be a place for their use in conjunction with regional anesthesia and whether there was a preferable route for NSAID administration. METHODS: Ilioinguinal and field blocks were performed preoperatively on seventy patients undergoing outpatient inguinal hernia repair. Patients were divided into a control group who received no ketorolac and four study groups who received a preoperative dose of 30 mg ketorolac by one of the following routes: i.v., i.m., p.o., or intrawound (i.w.). The ketorolac in the i.w group was mixed in the syringe with the local anesthetic used for the field block. i.v. and i.m. groups also received ketorolac at the time of the preoperative regional anesthesia and the PO group received the dose at least one hour preoperatively. All patients received a similar general anesthetic intraoperatively. RESULTS: Postoperative pain scores and analgesic requirements were lowest for the i.m., i.v., and i.w. groups. Pain scores and analgesic requirements for the PO group were less than for the control group but more than for the other three groups. Analgesic efficacy therefore ranked: i.m. = i.v. = i.w. > p.o. > control. Though no differences were found between groups in the time to discharge from the recovery room, the ease of nursing care paralleled the findings for pain scores and analgesia requirements. CONCLUSION: Beyond the analgesia provided by the regional anesthesia of the ilioinguinal and field blocks, the preoperative use of ketorolac further reduced postoperative pain scores and the need for additional postoperative analgesic medication. Comparable outcomes for the i.v., i.m. and i.w. groups indicate the lack of any benefit to concentrating the non-steroidal anti-inflammatory drug at the wound (i.w.) or to achieving high blood levels rapidly (i.v.). In conclusion, ketorolac is a useful supplement to ilioinguinal plus field block regional anesthesia for hernia surgery and is most effective administered parenterally. PMID- 8721469 TI - Treatment of acute reflex sympathetic dystrophy with DMSO 50% in a fatty cream. AB - Acute Reflex Sympathetic Dystrophy (acute RSD) was defined using a reproducible classification. Elevated temperature of the affected extremity ("calor"), measured by the dorsal side of the observer's hand and mentioned by the patient, pain ("dolor") measured by the Visual Analogue Scale (VAS), redness ("rubor"), edema ("tumor") and limited active range of motion ("functio laesa"), all contributed to the classification system. Patients scoring 4 to 5 positive symptoms were considered to have acute RSD. A prospective, randomized and double blind study was performed in 32 patients, all suffering from acute RSD. In all of these patients the primary injury was the result of a previous accident. One patient was taken out of the study because of his surgery. The study involved treatment with a fatty cream with 50% dimethyl sulfoxide (DMSO, group A), or without DMSO (placebo, group B), both for 2 months. All patients received physiotherapy applied within pain limits. Application of the creams resulted in both groups in an improvement of RSD-scores and VAS-scores after 2 months. However, the improvement of the RSD score in patients of group A (DMSO-group) was significantly (P < 0.01) better compared to group B. The results suggest a certain activity of DMSO 50% cream in patients suffering from RSD and is, therefore, recommendable. PMID- 8721470 TI - Menstrual stage influences postoperative nausea and vomiting following epidural buprenorphine. AB - BACKGROUND: Although buprenorphine sometimes causes severe emesis, its relation to the menstrual cycle has not been reported. METHODS: We conducted a prospective study on 68 reproductive-age women following lower extremity surgery under epidural anesthesia plus buprenorphine, to assess the effect of the day of the menstrual cycle on the incidence of postoperative nausea and vomiting. The patients were divided according to the phase of the menstrual cycle into four groups: day 1-7 group, day 8-14 group, day 15-24 group and day 25 to end of the cycle group. RESULTS: Nausea and vomiting were reported in 46 patients (67.6%), and the incidence was higher in the day 25 to end of the cycle group than in the day 8-14 or day 15-24 groups, and higher in the day 1-7 group than in the day 8 14 group. CONCLUSION: We conclude that emesis after epidural buprenorphine is related to the menstrual cycle. PMID- 8721471 TI - Effect of P-6 acupressure on prevention of nausea and vomiting after epidural morphine for post-cesarean section pain relief. AB - BACKGROUND: Nausea and vomiting are important side effects following administration of epidural morphine for post-Cesarean section pain relief. Stimulation of the P-6 (Neiguan) acupoint is a traditional Chinese acupuncture modality used for antiemetic purpose; it has been found to be effective. The aim of this study was to evaluate the antiemetic effect of P-6 acupressure in parturients given epidural morphine for post-Cesarean section pain relief. METHOD: In a randomized, double-blind and controlled trial, sixty parturients receiving epidural morphine for post-Cesarean section pain relief were investigated. Parturients were allocated to receive the acupressure bands or placebo bands on the P-6 acupoint bilaterally before the administration of spinal anesthesia and were observed over a 48-hour study period. RESULTS: The incidence of nausea and vomiting was significantly decreased from 43% and 27% in the control group, to 3% and 0% in the acupressure group, respectively (P < 0.05). CONCLUSION: The results demonstrate that prophylactic use of acupressure bands bilaterally on the P-6 acupoint can significantly reduce incidence of nausea and vomiting after epidural morphine for post-Cesarean section pain relief. PMID- 8721472 TI - Surgery during inhalation with nitric oxide in a patient with severe adult respiratory distress syndrome (ARDS). AB - Nitric oxide (NO) is increasingly used in intensive care units (ICU) in order to treat serious hypoxaemia secondary to ARDS. Since interrupting NO delivery in such patients for more than a few minutes could lead to serious adverse events, moving the patient outside the ICU has been very difficult. Recently developed equipment (Noresc 1503) enabled us to deliver 10 or 20 ppm NO from a ventilatory bag with reservoir. This is used for ventilation under transport from the ICU to the operating theatre (OT) or radiology department. We present a patient with severe ARDS undergoing major surgery while being treated with NO. The patient could be moved to the OT and operated on during five hours without significant changes in vital functions. PMID- 8721473 TI - Cardiac failure aggravated by timolol eye drops: preoperative improvement by changing to pilocarpine. AB - A 73-year-old woman with cardiac dysfunction had several episodes of severe bradycardia and pulmonary oedema when waiting for peripheral vascular surgery. She used timolol eye drops for primary open-angle glaucoma. The first episode of pulmonary oedema occurred two weeks prior to and the second on the day before the planned surgery. There were another two episodes of pulmonary oedema before she was transferred to the Department of Internal Medicine where she had a further two episodes of cardiac failure. After changing timolol to pilocarpine eye drops, the patient's condition was stabilized, and two weeks later surgery and postoperative recovery were uneventful. PMID- 8721474 TI - Pulmonary edema following intrauterine methylene blue injection. AB - Methylene blue is a commonly used dye in diagnostic procedures such as fistula detection, delineation of body tissues during surgery and for checking the patency of fallopian tubes. Many such dyes interfere with accurate measurement of oxygen saturation by a pulse oximeter. We report here a case in which methylene blue hot only interfered with pulse oximetery but also caused pulmonary edema in an ASA grade 1 patient. PMID- 8721475 TI - Management of upper airway obstruction using a Seldinger minitracheotomy kit. AB - Two adult patients with upper airway obstruction due to epiglottitis and infectious mononucleosis requiring urgent airway management are presented. Successful establishment of an open airway was achieved by performing a minitracheotomy using the Seldinger technique followed finally with nasotracheal fibreoptic intubation. PMID- 8721476 TI - Gangrene and renal failure caused by dihydroergotamine used to treat raised intracranial pressure following head trauma. AB - A case of peripheral gangrene and renal failure following the use of dihydroergotamine (DHE) for treatment of raised intracranial pressure (ICP) is reported. There have been several reports of vasospasm and peripheral gangrene caused by DHE used in combination with heparin for prevention of deep venous thrombosis. Clinical experience with the use of DHE in head trauma is limited. This case report emphasises the seriousness of the adverse effects associated with the use of this drug. PMID- 8721477 TI - Nitrous oxide and cerebral haemodynamics. PMID- 8721478 TI - Shivering during epidural anesthesia. PMID- 8721479 TI - Elevated levels of circulating intercellular adhesion molecule-3 (cICAM-3) in Psoriasis. AB - Intercellular adhesion molecule (ICAM)-3 is important in regulating leukocyte function and T-lymphocyte-antigen presenting cell interactions. Soluble, circulating forms of ICAM-3 and ICAM-1 (cICAM-3, cICAM-1) exist in serum, and levels are elevated in a variety of autoimmune diseases. Two types of soluble circulating tumour necrosis factor receptor (cTNF-R1, cTNF-R2) are found in the sera of healthy people. cTNF-R1 binds TNF-alpha and is important in regulating TNF-alpha-mediated inflammation. Psoriasis is a T-lymphocyte-mediated disease, characterized by cutaneous expression of ICAM-1, ICAM-3 and TNF-alpha. As it is unknown whether cICAM-3 is increased in sera of patients with psoriasis, we measured serum levels of cICAM-3 and compared them to levels of cICAM-1, cTNF-R1 and clinical severity of psoriasis. Sera was taken from 112 healthy controls and 32 patients with psoriasis. Clinical severity of psoriasis was assessed using the psoriasis area severity index (PASI). cICAM-1, cICAM-3 and cTNF-R1 in serum were quantitated using a dual antibody, solid phase ELISA, Levels of cICAM-3, cICAM-1 and cTNF-R1 were significantly increased in sera of patients with psoriasis as compared with controls, and these elevated levels correlated with clinical severity of psoriasis as assessed by the PASI. Also, there were good correlations between serum levels of cICAM-3, cICAM-1 and cTNF-R1 in psoriasis. These results demonstrate, for the first time, that circulating levels of cICAM-3 are increased in psoriasis and that these levels correlate both with disease severity and with elevated levels of cICAM-1 and cTNF-R1. The exact physiologic roles of circulating, soluble adhesion molecules and cTNF-R1 are unknown, but it is hypothesised that elevation of their circulating levels, as observed in psoriasis, may play a role in modulating the inflammatory reactions occurring in this disease. PMID- 8721480 TI - Response of the clinically uninvolved skin of psoriatic patients to tape stripping during acitretin treatment. AB - The aromatic retinoids etretinate and acitretin are widely used in the systemic treatment of severe psoriasis. The purpose of the present investigation was to further elucidate the mode of action of acitretin on abnormal keratinization and epidermal hyperproliferation in an in vivo model. Studies on the interference of acitretin with epidermal hyperproliferation and abnormal keratinization in psoriatic plaques are difficult to interpret, as acitretin-induced changes might be due to direct effects of acitretin or be the indirect effect of retinoid induced modulation of cutaneous inflammation. Using an immunohistochemical assessment, we examined the in vivo effect of systemic acitretin ( > 35 mg daily) on the expression of filaggrin, involucrin, and on the recruitment of cycling epidermal cells, in the tape-stripped uninvolved skin of psoriatic patients, a model which provides the opportunity to study epidermal regeneration in the absence of significant accumulation of T-lymphocytes. During acitretin therapy and 3 weeks after withdrawal of acitretin, we took biopsies from uninvolved skin following tape-stripping in 6 patients with psoriasis. Six patients with psoriasis who had never used acitretin served as controls. We did not observe a Koebner response in our patients after tape stripping. Filaggrin expression was decreased, while the recruitment of cycling epidermal cells and the involucrin expression were increased in the biopsies taken from patients who did not use acitretin. During acitretin treatment, however, the filaggrin expression was similar, whereas the Ki-67 positive nuclei and the involucrin expression showed a statistically significant decrease compared to those parameters in the patients who did not use acitretin. Our findings indicate that epidermal hyperproliferation and abnormal keratinization are modulated directly by acitretin. PMID- 8721481 TI - Soluble Pityrosporum-derived chemoattractant for polymorphonuclear leukocytes of psoriatic patients. AB - The chemoattraction of polymorphonuclear leukocytes (PMNs) from psoriatic patients, atopic patients and healthy control persons by Pityrosporum orbicularelovale was investigated using the Boyden chamber method. The chemotactical attraction of PMNs from psoriatic patients by Pityrosporum (stimulation index SI = 58 +/- 50) was significantly increased (p < 0.05) compared to PMNs from atopic patients (SI = 20 +/- 17) and control persons (SI = 26 +/- 24). This effect seems to be specific for Pityrosporum, since the chemotactical response to Staphylococcus epidermidis was not increased in psoriasis. The chemotactical factor produced by Pityrosporum is hydrophilic and is destroyed by acid hydrolysis, indicating its protein nature. The yeast Pityrosporum may thus play a role in the koebnerization of psoriasis. PMID- 8721482 TI - The membrane attack complex of complement alters the membrane integrity of cultured endothelial cells: a possible pathophysiology for immune complex vasculitis. AB - Recently, the possibility of direct involvement of the membrane attack complex of complement in the endothelial damage of immune complex vasculitis has been pointed out. However, no studies have so far elucidated this mechanism. The present study investigated the effects of complement on the membrane integrity of endothelial cells, using the fluorescein diacetate and ethidium bromide staining method. Cultured human umbilical vein endothelial cells were maintained in medium containing 10% zymosan-activated normal human serum. Cell detachment began to occur after 3 h of incubation, and the number of fluorescein diacetate-positive adherent cells decreased significantly, whereas that of ethidium bromide-positive detached cells increased significantly. Heat inactivation of the serum or replacement of the complement source with non-activated normal human serum or C5 , C7- or C9-deficient serum resulted in complete inhibition of these effects. These results suggest that complement induces detachment of endothelial cells by altering the cell membrane integrity and support the contention that the membrane attack complex of complement plays a significant role in the mechanisms of endothelial cell damage in immune complex vasculitis. PMID- 8721483 TI - Growth factor mRNA levels in alopecia areata before and after treatment with the contact allergen diphenylcyclopropenone. AB - The early immune response in alopecia areata is characterized by a Th1 T helper cell cytokine pattern and an aberrant expression of ICAM-1 and HLA-DR molecules on lesional hair bulbs. A counteracting cytokine pattern induced by a therapeutic contact dermatitis is supposed to mediate the hair regrowth. In addition to cytokines, growth factors have been shown to influence immune responses, and we therefore investigated the expression levels for a panel of growth factors in untreated versus alopecia areata after treatment with the contact sensitizer diphenylcyclopropenone. Using semiquantitative reverse transcriptase polymerase chain reaction we detected a striking overexpression of transforming growth factor beta 1 mRNA in successfully treated patients. This cytokine has been shown to be a potent immune response modifier, which can suppress Th1 immune responses. The way in which topical immunotherapy induces hair regrowth in alopecia areata is unknown, but a lesional increased expression of transforming growth factor beta 1 may be a possible mechanism. PMID- 8721484 TI - Serum cytokine and anti-Fc gamma R autoantibody measurements in patients with systemic sclerosis. AB - The serum IL-1 beta, IL-2, IL-4, IL-6, IL-8, TNF-alpha and soluble IL-2 receptor levels were measured, and the presence of anti-Fc gamma receptor (Fc gamma R) antibodies was investigated in the sera of 18 patients with systemic sclerosis (SSc). An increase of TNF-alpha was detected in 8 of the 18 cases. II-1 beta was elevated in all the 18 patients. Both IL-2 and IL-4 were elevated in 7 cases. The IL-6 level was elevated in 17 patients, while IL-8 was increased in all cases. The soluble IL-2 receptor level was elevated in 11 patients. Fc gamma R-specific antibodies were detected in the sera of 6 patients, and there was a significant association between anti-Fc gamma R antibody positivity and IL-4 elevation. The presence of anti-Fc gamma R antibodies may influence several cell functions and may contribute to the remarkable variability of cytokine levels in SSc. PMID- 8721485 TI - Integrin molecules: a clue to the non-metastasizing behaviour of basal cell carcinomas? AB - Does the integrin profile of basal cell carcinomas explain their non metastasizing behaviour? Immunohistochemical Investigation of nodular (n = 31) and superficial (n = 17) tumours yielded a strong expression of alpha 2, alpha 3, alpha 6, beta 1, and beta 3 subunits and a weak expression of alpha 4 subunits by the epithelial tumour component. alpha 5 subunits were focally detected in superficial basal cell carcinomas but not in the nodular type. Tumour cells were nearly devoid of alpha v subunits. The integrin profile of basal cell carcinomas does not differ essentially from that of metastasizing tumour varieties and cannot be regarded as a major reason for the non-metastasizing phenotype of basal cell carcinomas. PMID- 8721486 TI - Cell-mediated immunity of patients who have had basal cell carcinomas. AB - The cell-mediated immunity of patients who had basal cell carcinomas (BCCs) removed was studied by measuring cuntaneous delayed hypersensitivity reactions to recall antigens (Multitest CMI, Pasteur-Merieux), and by measuring lymphocyte counts and subsets. One group of patients had multiple BCCs (3 or more) removed and were considered to have a high risk of new BCC formation. The other group consisted of patients who had one BCC and had not developed another within 5 years; these were considered to have a low risk of new BCC formation. The low risk patients had significantly larger cutaneous reactions to recall antigens (p < 0.05) and significantly fewer were anergic (p < 0.01). There was a correlation between smaller cutaneous reactions and increasing numbers of BCCs (p < 0.05). There was no significant difference between the groups in lymphocyte counts or subsets, but the low-risk patients had a significantly higher CD4:CD8 ratio (p < 0.05) than the high-risk group. The Multitest CMI test can be used to determine which patients are at risk of developing many BCCs. PMID- 8721487 TI - Sun exposure and sunscreen use among sunbathers in Denmark. AB - Eight hundred and eight sunbathing Danes were interviewed in order to register the duration of sun exposure and sunscreen use. The mean sun exposure time, 203 min, was found to be independent of sex, age and use of sunscreen. Overall, sunscreens were used by 65%, 73% of the females and 52% of the males. The median sun protection factor used by subjects older than 10 years was 5; in the case of children 10 years or younger, it was 12. Eight per cent of the interviewed subjects used more than one factor. Fifty-seven per cent of the sunscreen users applied the sunscreen prior to arriving at the beach. The sun exposure time was not significantly different in the group of subjects using sunscreens, compared to the group of subjects not using sunscreens. This study indicates that sunscreen users may not increase their exposure to the sun. PMID- 8721488 TI - Stage-dependent expression of CD7, CD45RO, CD45RA and CD25 on CD4-positive peripheral blood T-lymphocytes in cutaneous T-cell lymphoma. AB - Flow cytometric analysis of T-cell surface markers in peripheral blood has revealed abnormal patterns in patients with cutaneous T-cell lymphomas (CTCL). Here we investigated CD7, CD25, CD45RO and CD45RA expression on CD4+ T lymphocytes in patients with CTCL stage I/II and III/IV and in patients with severe inflammatory skin diseases (ISD), as well as in healthy controls. Only late stage CTCL (III/IV) showed a lymphocytosis with a distinct surface marker pattern: CD3+, CD4+, CD8-, CD7-, CD45RO+, CD45RA-. Early stage CTCL (I/II) showed normal lymphocyte counts, a normal T-helper cell expression of CD7, CD45RO and CD45RA, and a slightly increased percentage of CD4+ CD25+ lymphocytes, which was also found in ISD. It is concluded that flow cytometric analysis of the above T cell surface markers may be useful in the diagnosis of patients with late stage CTCL. However it does not allow us to distinguish patients with early stage CTCL from patients with ISD or controls. PMID- 8721489 TI - Appearance of Sezary-like atypical lymphocytes in the regressing lesions of juvenile xanthogranuloma. Its role in the spontaneous regression. AB - A case of juvenile xanthogranuloma of the skin was sequentially biopsied for 10 months. In the electron microscopic examination of the regressing lesions, we observed that cells with highly indented nuclei, Sezary-like cells, were in close apposition to vacuolated degenerating histiocytes in many foci. In immunohistochemical stain, the number of UCHL-1+ cells were increased in regressing lesions compared with early lesions. We speculate that these Sezary like atypical lymphocytes may be closely related to the spontaneous regression of juvenile xanthogranuloma. PMID- 8721490 TI - Multiple keratoacanthomas, giant keratoacanthoma and keratoacanthoma centrifugum marginatum: development in a single patient and treatment with oral isotretinoin. AB - A 78-year-old man is described, who over 18 years developed three different types of keratoacanthoma: multiple keratoacanthomas, keratoacanthoma centrifugum marginatum and giant keratoacanthoma. Histological examination of the different neoplasms showed similar changes, all typical of a keratoacanthoma. In situ hybridisation revealed no human papilloma virus in the tumours. Complete examination showed no associated internal malignancy. After repeated surgical treatment oral isotretinoin treatment was administered (1 mg/kg per day). This treatment produced clearing of existing keratoacanthomas and, during a period of 2 months, further keratoacanthoma formation was completely suppressed. Treatment was stopped after 3 months by the patient because of side-effects. Numerous keratoacanthomas developed during the following 6 weeks. PMID- 8721492 TI - Pruritus circumscriptus sine materia: a sequel of postzosteric neuralgia. Evaluation by quantitative psychophysical examination and laser-evoked potentials. AB - A case of circumscribed pruritus existing since 1 year on clinically uninvolved skin is reported, in which careful history revealed a 5-year previous episode of herpes zoster in the same dermatome. Impairment of cutaneous sensitivity was evaluated by use of a quantitative psychophysical examination and laser-evoked cortical potentials (LEP). PMID- 8721491 TI - Treatment of chronic idiopathic urticaria with topical steroids. An open trial. AB - Twenty-three patients with chronic idiopathic urticaria and 3 patients with urticaria pigmentosa received a 2-week treatment with daily application of potent topical steroid using plastic occlusion. Seventy per cent of the chronic urticaria patients had a statistically significant, immediate response, which, however, did not last until the final evaluation after 1.5 years. Relapse occurred after on average 3 weeks, but 39% reported less severity. The 3 patients with urticaria pigmentosa remained free from attacks for 6 to 9 months, after which they gradually relapsed. Two women were diagnosed during the follow-up period to have a carcinoma of the breast, and one patient developed systemic lupus erythematosus. PMID- 8721493 TI - Two-point discrimination of itch in patients with atopic dermatitis and healthy subjects. AB - The ability to perceive two pruritic stimuli as separate depending on the distance between them (two-point discrimination of itch) was determined in a single-blind study of 20 patients with atopic dermatitis and 20 healthy subjects. Itch was induced with pairs of histamine injections (0.1 micrograms each) given on the upper arm, either along its axis (longitudinally within a dermatome) or at right angles to it (transversally involving more than one dermatome). The interinjection distances were varied in 3-cm steps until the shortest distance at which the two itching stimuli could be perceived as separate was found. Pairs consisting of one injection of histamine (0.1 micrograms) and one of saline served as controls. The two-point discrimination of itch was significantly better in the atopic dermatitis patients than in the healthy controls, both longitudinally (atopic dermatitis: median 12 cm, range 3-21 cm; healthy controls: 15 cm, range 9-21; p < 0.01) and transversally (atopic dermatitis: median 6 cm, range 3-18; healthy controls: 12 cm, range 6-21; p < 0.001). The two-point discrimination of itch did not correlate with the subject's age, two-point discrimination of touch, or, for the atopic dermatitis patients, with their eczema scores or serum IgE levels. The quality, intensity or spatial summation of itch did not differ significantly between the two groups of subjects. PMID- 8721494 TI - Lack of sympathetic involvement in dermatitis confined to the median nerve territory. A case report. AB - Both decreased and increased sympathetic nerve activity has been suggested as a possible underlying mechanism in inflammatory skin lesions. Modulation of sympathetic function has been proposed in the treatment of dermatitis. This case report describes the investigation strategy and normal findings in a case of dermatitis strictly confined to the median nerve territory, illustrating the need for specific tests of sympathetic function when pharmacological as well as physical sympatho-modulatory therapies are considered. PMID- 8721495 TI - Flare-up at contact allergy sites in a gold-treated rheumatic patient. AB - Contact allergy to gold sodium thiosulfate and gold sodium thiomalate was established by skin testing in a rheumatic patient intended for gold therapy. An intramuscular test dose of gold sodium thiomalate (Myocrisin) induced a flare-up of previously positive epicutaneous and intradermal test reactions, with a histological and immunohistochemical picture compatible with an allergic contact dermatitis. Since gold allergy is frequent, the cutaneous side-effects of gold therapy ("gold dermatitis") may be explained by such an immunopathological reaction. PMID- 8721496 TI - Familial Kaposi's sarcoma: case reports and review of the literature. AB - We report 4 new families with Kaposi's sarcoma, occurring in 2 members of the same family (2 pairs of brothers, uncle and nephew and father and son) in a series of 160 patients with Kaposi's sarcoma. In addition, HLA typing was also carried out for 6 of the 8 patients. A detailed review of the literature is also presented. The total number of familial cases of Kaposi's sarcoma is low (only 30 cases are described); the great majority of these cases consist of siblings and Italians. PMID- 8721497 TI - Brittle and sparse hair with normal cystine content caused by methionine deficiency? AB - The unusual case of an 8-year-old girl with a hair disorder is presented, characterized by brittle, short and sparse hair. On polarizing microscopy the latter reveals a "tiger tail" pattern, whereas severe cuticular defects are detected on scanning electron microscopy. The patient's hair has a normal cystine content but is completely devoid of methionine and reveals distinct changes of its visoelastic parameters. It is presently unknown whether the lack of methionine may be implicated in the pathogenesis of this hair disorder, which to the best of our knowledge has not been previously described. PMID- 8721498 TI - A therapeutic approach to erythrodermic psoriasis: report of a case and a discussion of therapeutic options. AB - In this case report a patient with therapeutically recalcitrant erythrodermic psoriasis is presented. After various attempts with several major therapies in this patient, the first substantial improvement was achieved using the combination of cyclosporine and calcipotriol, followed by the combination of UVB and calcipotriol. The therapeutic options for severe psoriasis are discussed, and since combined approaches seem to be an attractive alternative for severe psoriasis, mechanisms of synergy of combined therapeutic approaches are hypothesised. PMID- 8721499 TI - Generalized pustular psoriasis in Japan: two distinct groups formed by differences in symptoms and genetic background. AB - A multi-center study for investigating generalized pustular psoriasis was carried out in Japan in order to clarify the prevalence, the etiology, and the standard therapy. Through questionnaires, 541 cases of patients with generalized pustular psoriasis and related disorders were collected, and the history, the precipitating factors, the symptoms and the laboratory findings were analyzed from medical records. The statistic analysis revealed that 902 (+/- 102.2) patients with generalized pustular psoriasis were expected at the time, and that the prevalence of generalized pustular psoriasis in Japan was 7.46/million. Of 541 patients with generalized pustular psoriasis, 208 cases had the recurrent episodes of symptoms with generalized pustules which had been referred to von Zumbusch type pustular psoriasis or to the acute form of generalized pustular psoriasis. Two hundred and eight patients were further subdivided into two groups: one with a preceding history of ordinary psoriasis (pso+ generalized pustular psoriasis; 65 cases) and another without a psoriasis history (pso- generalized pustular psoriasis; 143 cases). Subdivision into the two groups defined that the onset of pustular outbreak was earlier in the pso- generalized pustular psoriasis group. It was also pointed out that the precipitating factors were different, as the pso- generalized pustular psoriasis group was more frequently affected after infections, and that the pso+ generalized pustular psoriasis group was more frequently affected by preceding corticosterold therapy. HLA analysis of 92 cases with von Zumbusch type generalized pustular psoriasis confirmed that A2, B14, B35 phenotypes were weakly correlated, and when 92 cases were subdivided into the two groups, the pso+ generalized pustular psoriasis group revealed a statistically significant correlation with A1 (p < 0.01), B37 (p < 0.02) and DRw10 (p < 0.05), which was closely related with psoriasis vulgaris patients in Japan. Both clinical surveillance and genetic analysis have disclosed the heterogeneity of von Zumbusch type generalized pustular psoriasis, as there exist two types; one is closely related to psoriasis and the other not. PMID- 8721500 TI - Morphologic features and clinical significance of skin involvement in patients with AIDS-related cryptococcosis. AB - Eight out of 30 consecutive patients with AIDS and cryptococcal meningoencephalitis (26,7%) presented cutaneous involvement, in the form of papulo-nodular lesions in 3 patients, molluscum contagiosum-like lesions in 3 patients, and pustular-ulcerative lesions in 2 patients. Skin localization represented an untoward prognostic sign, and in 3 out of 8 patients the lesions were diagnosed 2 to 6 weeks before the onset of systemic and neurologic signs and symptoms of the disease. A systematical examination of all suspected cutaneous lesions in subjects with advanced HIV disease may lead to an earlier diagnosis and treatment of disseminated cryptococcosis. PMID- 8721501 TI - Scrotal angiokeratoma in a young man. PMID- 8721502 TI - A possible role of interleukin-8 in the induction of psoriasis-like lesions in Torre-Muir syndrome. PMID- 8721504 TI - Recurrent aphthous stomatitis: treatment with pentoxifylline. PMID- 8721503 TI - Clofazimine in inflammatory facial dermatosis--granuloma faciale and lipogranulomatosis subcutanea (Rothmann-Makai) PMID- 8721505 TI - Pretibial epidermolysis bullosa with vulvar involvement. PMID- 8721506 TI - Intralesional bleomycin for the treatment of non-genital warts in HIV-infected patients. PMID- 8721507 TI - Lateral edge nail involvement indicates poor prognosis for treating onychomycosis with the new systemic antifungals. PMID- 8721508 TI - Discoloration of the nail plate due to the misuse of amorolfine 5% nail lacquer. PMID- 8721509 TI - Immunological features of chronic adult paracoccidioidomycosis: report of a case treated with fluconazole. PMID- 8721510 TI - Cutaneous Horn in a lesion of prurigo nodularis. PMID- 8721511 TI - Wound healing with a new growth factor formula. PMID- 8721512 TI - Discoid lupus erythematosus during treatment with cyclosporine. PMID- 8721513 TI - Milker's nodule--a report of 15 cases in the county of North Jutland. PMID- 8721514 TI - Cold urticaria and delayed pressure urticaria in the same patient. PMID- 8721515 TI - Occupational allergic contact dermatitis from epoxy resin in a dental nurse with primary sensitization during cyclosporine treatment. PMID- 8721516 TI - Reactivation of healed primary syphilitic ulcer: a manifestation of febrile Herxheimer reaction in secondary syphilis. PMID- 8721517 TI - Cytokine profile of bilateral pseudocyst of the auricle. PMID- 8721518 TI - Ofuji's papuloerythroderma: report of a new case responding to PUVA. PMID- 8721519 TI - Clinical and serological manifestations of genital human papillomavirus infection. AB - Efficacy of chemical and/or surgical treatment for penile and anal condylomata acuminata was investigated in two retrospective studies of hetero- and homosexual men. Variation in clinical features and symptomatology as well as the reliability of diagnostic criteria by different methods for acetowhite penile lesions was also studied. Furthermore, the antibody response in the course of penile wart disease as well as in asymptomatic genitoanal papillomavirus infection (GPVI) was analysed. In the first retrospective study, as much as 23% of patients still had condylomas after one year of chemical and/or surgical treatment. On the other hand, 38% were cured after a single treatment session. In the group mainly with anal warts, concurrent penile warts were significantly more common among heterosexual men compared to homosexual men (p < 0.001), while intra-anal wart growth was more common among the homosexual males (p < 0.001). When comparing diagnostic methods for subclinical penile HPV infection, conventional histopathology appeared to be the most valuable diagnostic aid to penoscopy, while the additional use of Southern blot, in situ hybridisation and PCR assays for HPV DNA detection did not increase the predictive value of GPVI. We also describe a new distinct clinical entity, HPV-associated balanoposthitis, comprising a wide range of often long-lasting symptoms, such as itching, burning and dyspareunia. A significant increase in the IgG antibody response against defined epitopes in the L1 and L2 capsid proteins of HPV 6, was found among men with previous condylomata. By following a cohort of STD clinic patients with multiple brush samples from the genitoanal region as well as serum samples taken at several consecutive clinical visits, we identified 16 patients who had seroconverted to HPV seropositivity during follow-up. Antibody responses to several HPV-derived peptide and protein antigens were induced at the same time. Seroconversions were usually seen concomitantly with HPV acquisition or at the visit after HPV DNA was first detected. The HPV antibody response was frequently transient and declined or disappeared after clearance of infection. The antibody responses were induced by several different HPV types, indicating limited type specificity. The most type-restricted response was against HPV 16 capsids, where seroconversions to continuous seropositivity were induced by infection with HPV 16. PMID- 8721520 TI - The coexistence of psoriasis with lupus erythematosus and other photosensitive disorders. AB - The coexistence of psoriasis with LE or other photosensitive disorders is rare in our patient population, occurring in 0.69% of patients with psoriasis and 1.1% of those with LE. PMLE was the most common cause of photosensitivity in psoriatic patients without LE, occurring in 32%. Less common causes included drug-related photosensitivity (thiazides and thiazide derivatives in four of the five cases), PUVA reactions, and photocontact reactions. The Goeckerman regimen or UVB applied in a cautious, well-controlled atmosphere was generally well tolerated in this group, including patients with PMLE. Photosensitivity occurred in 50% of our patients with psoriasis and LE, and it was secondary to LE in 70% of cases. Most patients were female and had SLE. Psoriasis developed first in 55% of the cases. Studies that were useful for distinguishing photosensitive from nonphotosensitive patients with SLE included determination of antibodies to extractable nuclear antigens (67% versus 14%), double-stranded DNA (64% versus 9%), and skin biopsy for direct immunofluorescence (58% versus 27%). Occasional patients have features suggestive of photosensitivity with or without signs or symptoms of LE. These patients may have atypical psoriatic plaques occasionally yielding routine histology diagnostic of psoriasis with direct immuno-fluorescence results suggestive of lupus. Frequently, connective tissue serology findings are positive, and affected patients require close follow-up for the development of LE. In patients with psoriasis and suspected photosensitivity, we recommend a careful history and examination, skin biopsy for routine histology and direct immunofluorescence, blood tests including determination of antibodies to antinuclear antibodies (Hep-2 substrate if negative on routine substrate), extractable nuclear antigens, and double-stranded DNA, and phototesting when indicated. Large-scale prospective studies are required before the most appropriate therapy for patients with psoriasis and LE can be recommended. PMID- 8721521 TI - Anticholinergic antimuscarinic agents in the treatment of airways bronchoconstriction in children. PMID- 8721522 TI - Monomeric chemically modified allergens: immunologic and physicochemical characterization. AB - Allergenic extracts (Der p, grass, and Parietaria) or single allergens such as Par j I (the major allergen of Parietaria) and ovalbumin (OA), a food allergen widely used in animal models, were chemically modified by reaction with potassium cyanate (KCNO), which transforms the epsilon-amino group of the lysine of proteinaceous allergens into ureido groups. KCNO-modified (carbamylated) allergens have low allergenic potency, as demonstrated in vitro (RAST inhibition) and in vivo (passive cutaneous anaphylaxis). When used to immunize rabbits, carbamylated allergens still induce IgG antibodies able to cross-react with native allergens (immunoblotting experiments). An interesting feature distinguishing carbamylated allergens from other chemically modified allergens is the preservation of the native monomeric dimension as demonstrated by SDS-PAGE analysis. Results are discussed from the perspective of clinical application of carbamylated allergens. PMID- 8721523 TI - The atopy trait in hypersensitivity to nonsteroidal anti-inflammatory drugs. AB - The prevalence of atopy was evaluated in two groups of subjects with hypersensitivity to nonsteroidal anti-inflammatory drugs (NSAID): 1) 78 patients with aspirin-induced asthma (AIA) confirmed by oral or bronchial provocation challenges 2) 42 subjects with hypersensitivity to pyrazolone drugs (case history and positive skin tests to noramidopyrine/aminophenazone) who tolerated aspirin well. Fifty sex- and age-matched persons from an unselected general population, with no hypersensitivity to NSAID, formed the control group. Atopy was estimated from the results of the following clinical and biologic parameters: 1) personal and family history of atopic diseases 2) skin prick tests with 16 aeroallergens 3) serum levels of specific IgE to five aeroallergens 4) total serum IgE level. Different definitions of atopy were used, consisting of constellations of two or three of the above-mentioned features. The results of the study revealed that the prevalence of atopy varied according to the criteria used for its definition. Irrespective of the definition used, a similar distribution of atopy was observed in both groups of patients with hypersensitivity to NSAID. Atopy was more frequent in either group of patients with intolerance of NSAID than in the control group. Thus, atopy is related to adverse drug reactions to NSAID. PMID- 8721524 TI - Histamine releasability of basophils and skin mast cells in chronic urticaria. AB - In order to clarify the pathogenetic role of basophils and mast cells in chronic urticaria, histamine and leukotriene (LT)C4 release was examined in washed mixed leukocytes (n = 8) and skin mast cells (n = 5) from patients with chronic urticaria and compared with the same cells from normal controls (n = 9). Anti-IgE stimulated basophil histamine release was significantly reduced in urticaria patients (median 2.9% vs 15.1% in normal controls), whereas histamine release to A23187, FMLP, and PAF, as well as anti-IgE-induced LTC4 release, showed no differences in both groups. In contrast, anti-IgE-stimulated skin mast cells from urticaria patients reacted similarly to those of controls (median histamine release 11.4% vs 14.2% in normal controls). Pretreatment of the cells with interleukin (IL)-3 upregulated responsiveness of basophil histamine release to anti-IgE in urticaria patients (median histamine release 14.3%), but pretreatment with the H2-antagonist cimetidine showed no effect. These data show that reduced basophil histamine releasability in chronic urticaria is not H2 mediated. It is a stimulus-, mediator-, and cell type-restricted phenomenon that can, at least partially, be reversed in the presence of the cytokine IL-3. PMID- 8721526 TI - Allergen avoidance in a day-care center. AB - The predominant sensitizing allergens in Swedish asthmatic children are furred pet animals. Poor ventilation and increased indoor humidity cause accumulation of pet allergens indoors. Indirect contacts with cats also increase pet allergen exposure. We investigated whether the concentration and the total amount of cat (Fel d I) and dog (Can f I) allergen in dust from various surfaces in a day-care center were influenced by extensive renovation, installation of a new ventilation system, ventilation of floors, and avoidance by families and personnel of direct and indirect contacts with pets. The cleaning routine in the day-care center was changed extensively after renovation. Old mattresses, pillowcases, curtains, sofas, and soft toys were removed, and new material were purchased. Dust was collected from various surfaces before and after renovation from table and chair surfaces, floors, curtains, mattresses, pillows, cushion-like toys, and sofas on five occasions during a 12-month period. Fel d I and Can f I allergen levels were determined by sandwich ELISA. The reduction in the total amount of both Fel d I and Can f I/sampling area was more pronounced than the reduction in the concentration of allergen/g of dust. After renovation, the mean cat and dog allergen concentration was decreased by six and 10 times, respectively. After 1 year, the total amount of cat allergen was lower in all areas (from 24 ng to < 1 ng/sampling area, P < 0.05). Already at the first sampling after renovation, the total amount of dog allergen had decreased more than 10 times. In a previous study, we found the median concentration of cat and dog allergen to be 10-30 times higher in ordinary day-care centers. Our present study demonstrates that children changing from a normal center to the renovated center would be exposed to much lower levels of cat and dog allergen. Combined measures such as installation of mechanical ventilation, ventilated floors, regular extensive cleaning, and, probably most important, avoidance of direct and indirect contacts with pets, should decrease cat and dog allergen exposure in day-care centers. PMID- 8721525 TI - Epitope analysis of HLA-DR-restricted helper T-cell responses to Der p II, a major allergen molecule of Dermatophagoides pteronyssinus. AB - T-cell epitopes of Der p II, a major allergen of Dermatophagoides pteronyssinus, were analyzed by using human T-cell clones. We tested 38 cloned T cells from two Japanese patients with allergic rhinitis, and identified at least two peptides (K33-T47 and I58-C73) as helper T-cell epitopes. The former epitope was shown to be restricted by HLA-DRB1*1502, and the latter by HLA-DRB1*0405, both of which are typical Japanese HLA-DR alleles, suggesting that those T-cell epitopes might be important for the onset of house-dust mite allergy in the Japanese population. We prepared 15 analog peptides of the HLA- DRB1*1502-restricted 15-mer peptide. Of those 15 residues, five (F35, L37, A39, F41, and E42) were critical for the epitope activity, and three residues (F35, A39, and E42) seemed to be included in anchor motifs for HLA-DRB1*1502. The epitope peptide was also recognized by HLA DRB1*1502-positive healthy donors; however, only allergic T cells showed Th2 functions. Antigen-presenting cells of nonallergic donors were able to activate allergic T cells to express Th2 function. This seemed to suggest that antigen recognition of T cells, as well as additional unknown factors which promote Th2, rather than Th1, responses, might be important for the onset of house-dust mite allergy. PMID- 8721528 TI - Allergic urticaria and angioedema caused by a hemostatic sponge of bovine fibrin used in tooth extraction. AB - We report the case of a 54-year-old woman with no history of atopic diseases or drug allergies who developed a generalized anaphylactic reaction with urticaria and angioedema a few hours after tooth extraction. The skin scratch and RAST tests demonstrated IgE-mediated hypersensitivity to the product Hemofibrine, a hemostatic sponge made of bovine fibrin. Of all RAST controls, only one serum with known positivity to bovine serum albumin (BSA) showed a low degree of positivity to Hemofibrine. Bovine fibrin should be added to the list of medical products that can elicit allergic reactions following dental treatment. PMID- 8721527 TI - Allergen-induced inflammation in the nose: a comparison of acute and repeated low dose allergen exposure. AB - To investigate allergic rhinitis induced by two experimental provocation models, we compared local inflammation with markers of eosinophil activity in peripheral blood. Patients with strictly seasonal allergic rhinitis were selected and investigated outside the pollen season. An acute challenge with increasing doses of allergen every 15 min until symptoms occurred was performed in nine patients. Nasal lavage and blood samples were taken before and 4 and 24 h after challenge. After a 6-week washout period, 10 patients were submitted to 7 days of repeated low allergen exposures. One small dose (approximately equal to 1/100 of the acute dose) was given each day. Blood and lavage samples were taken prior to and after the period. As control four patients were challenged with diluent only. The acute challenge resulted in sneezing and nasal discharge and blockage and was accompanied by a rise in histamine and eosinophil cationic protein (ECP) in lavage fluid after 4 h and continuing after 24 h, when there also was a rise in the number of eosinophils and ECP in peripheral blood. The repeated low-dose exposures caused very few symptoms but produced increased ECP in the lavage fluid and a trend toward increased histamine concentration. There were no changes in ECP, intracellular EG2 binding, or number of eosinophils in the blood. No changes were seen in the control group. Our findings show that changes in eosinophil mediator release in nasal lavage can be seen after very low, but repeated, allergen exposures despite no, or minimal, clinical symptoms. PMID- 8721529 TI - Effect of ethanol on airway caliber and nonspecific bronchial responsiveness in patients with alcohol-induced asthma. AB - No study has investigated the effects of ethanol on bronchial responsiveness in patients with alcohol-induced asthma, although acetaldehyde, which is a metabolite of ethanol and is thought to be a main factor in alcohol-induced asthma, causes both bronchoconstriction and bronchial hyperresponsiveness. The purpose of this study was to investigate the direct action of ethanol on the airway in patients with alcohol-induced asthma. First, we investigated the bronchial response to inhalation of ascending doses (5, 10, and 20%) of ethanol in nine patients with alcohol-induced asthma. Then, the bronchial responsiveness to methacholine was measured in 14 patients who were pretreated with saline or 20% ethanol in a double-blind, randomized, placebo-controlled, crossover fashion. Ascending doses of inhaled ethanol caused no significant changes in FEV1. The methacholine concentrations producing a 20% fall in FEV1 (PC20-MCh) after 20% ethanol (0.769 mg/ml, GSEM 1.514) were significantly (P = 0.0357) higher than those after saline (0.493 mg/ml, GSEM 1.368). This indicates that ethanol has a reducing effect on nonspecific bronchial responsiveness in patients with alcohol induced asthma; this paper is the first report on the effects of ethanol on bronchial responsiveness. PMID- 8721530 TI - Occupational conjunctivitis associated with type IV allergy to methacrylates. AB - On rare occasions, exposure to methacrylates (MAs) may induce mucosal symptoms. Three patients, two dental laboratory workers and one hearing aid laboratory worker, are presented. All three had allergic contact dermatitis from MAs which disappeared after avoidance of contact with uncured MA compounds. Two of the patients, the dental laboratory assistant and the hearing aid worker, had also developed symptoms of conjunctivitis. Both were exposed to chemically curable and light-curable MAs. The association between their conjunctivitis and type IV allergy to MAs was supported by the following observations: 1) sensitization to several MAs including methylMA, 2-hydroxyethylMA, ethyleneglycol diMA, triethyleneglycoldiMA, tetrahydrofurfuryldiMA, and 1,4-butanedioldiMA, as well as to pentaerythritol triacrylate; 2) simultaneous appearance of their eye symptoms and allergic contact dermatitis; 3) high exposure to MAs because of disturbances of ventilation; 4) disappearance of the eye symptoms during holidays; and 5) opthalmologist's findings of follicular conjunctivitis with some papillae, eosinophilia, and lymphocytosis in conjunctival scrapings corresponding to allergic contact conjunctivitis. In addition, the activation of eosinophils in the conjunctival scrapings was demonstrated with the monoclonal antibody technique, and an elevated level of eosinophilic cationic protein was found in the tear fluid. Our cases suggest that conjunctivitis may be caused by type IV allergy to MAs, although type I allergy (even though prick tests were negative), other hypersensitivity mechanisms, or irritation cannot be excluded. 4H-Gloves seem to give adequate hand protection even to allergic patients. More attention should be paid to ventilation systems if MA exposure occurs. In some cases, patients with eye symptoms need to be patch tested. PMID- 8721531 TI - The risk of general reactions to skin prick testing (SPT) PMID- 8721532 TI - Severe adverse reactions during specific subcutaneous immunotherapy. PMID- 8721533 TI - Allergic contact dermatitis from estradiol in a transdermal therapeutic system. PMID- 8721534 TI - Immediate-type allergy to the hair dye basic blue 99 in a hairdresser. PMID- 8721535 TI - Prevention of latex allergy. PMID- 8721536 TI - Improper treatment of anaphylaxis. PMID- 8721537 TI - Allergic contact dermatitis to chloramphenicol. PMID- 8721538 TI - Occupational hypersensitivity to sesame seeds. PMID- 8721539 TI - Consensus symposium on combined antiviral therapy. International Society for Antiviral Research and the National Institutes of Allergy and Infectious Diseases. PMID- 8721540 TI - The implications of drug resistance for strategies of combination antiviral chemotherapy. PMID- 8721541 TI - Combination therapy for HIV-1 infection-overview: preclinical and clinical analysis of antiretroviral combinations. PMID- 8721542 TI - Analysis of drug-drug interactions: an overview. PMID- 8721543 TI - Combination antiviral therapy for respiratory virus infections. AB - A limited number of antiviral drug combinations have been shown to have enhanced activity for important human respiratory viruses. Rimantadine or amantadine combined with ribavirin shows increased antiviral effects in vitro and in experimental animal models. This combination warrants testing in human influenza. Immunoglobulin containing neutralizing anti-RSV antibody combined with ribavirin shows enhanced antiviral effects in experimental animal infections and provides clinical benefit in severe RSV infections of transplant patients. Generally, more effective treatments for acute respiratory viral infections will likely involve combinations of both antivirals and agents that modulate host inflammatory responses to infection. PMID- 8721544 TI - In vitro evaluation of combination therapies against hepatitis B virus replication. AB - The HBV-producing human hepatoblastoma cell line, 2.2.15, has been shown to be an accurate model of chronic cellular viral infection and a predictive model of antiviral response for in vivo hepadnaviral infection. Our laboratory has utilized the 2.2.15 cell line in a standardized assay to examine treatment schemes which use combinations of clinically relevant nucleoside analogues, novel methods to deliver potentially useful nucleoside combinations, and treatments which simultaneously target different parts of the HBV replication pathway. For example, the combination of 3TC (lamivudine) with either alpha interferon or penciclovir significantly enhances the antiviral effectiveness of these agents against HBV replication in 2.2.15 cell culture. PMID- 8721545 TI - In vitro antiviral activity of 141W94 (VX-478) in combination with other antiretroviral agents. AB - 141W94 (VX-478) is a novel HIV-1 protease inhibitor with an IC50 of 0.08 microM against HIV-1 (strain IIIB) and a mean IC50 of 0.012 microM against six HIV clinical isolates. 141W94 was synergistic on the basis of isobologram analysis with each of the following reverse transcriptase inhibitors: AZT, 935U83, 524W91, 1592U89 and ddl, 141W94 was also synergistic with saquinavir and additive with either indinavir or ritonavir. Resistance to 141W94 has been reported in vitro passage experiments. The binding of 141W94 to human alpha 1-acid glycoprotein was relatively weak (Kd = 4 microM) and the off-rate for the drug is very fast (> or = 100 s-1). Only a 2-fold reduction of in vitro antiviral activity was observed in the presence of 45% human plasma. No serious drug associated adverse experiences were reported in a Phase I placebo-controlled, single-dose escalation, pharmacokinetic and safety study. The average concentration of 141W94 at 8 and 12 h after single doses of 900 and 1200 mg, respectively, was in excess of 10 times the IC50. As 141W94 is synergistic with a variety of anti-HIV-1 agents and exhibits a unique cross resistance profile compared to other protease inhibitors, 141W94 is considered a good candidate for combination therapy. PMID- 8721546 TI - Role of animal models in selecting antiviral combinations for clinical studies. AB - Although experimental viral infections in animals have been used extensively in the development of antiviral drugs used as monotherapy, they have not been utilized widely for evaluation of combination chemotherapy. One of the major reasons for the lack of use of animal models is that for the diseases that are the main target for combination therapy, AIDS and hepatitis B and C infections, there is a lack of suitable models for these diseases. In contrast, most combination studies in animal models have been directed against herpes simplex virus infections but there are relatively few patients available who would benefit from combination therapy over single agent therapy. In between those two extremes are the cytomegalovirus infections. While there are animal models available that have been predictive of efficacy in humans and there are sufficient patients available, the use of antiviral combinations in animal models and in humans have begun only recently. At the present time there is not enough information available to establish the predictability for any of the animal models for efficacy of combinations of antiviral agents. PMID- 8721547 TI - Combinations of ganciclovir and antibody for experimental CMV infections. AB - The effect of combined treatment with ganciclovir and hyper immune serum (HIS) was evaluated in three animal models. It concerned a generalized CMV infection model, a meningo-encephalitis model and an interstitial lung disease (ILD) model in immunocompromised rats. In the generalized model, the ganciclovir and HIS had a moderate synergistic effect on survival and greatly decreased virus titers in internal organs. In contrast, in the meningoencephalitis model, combined treatment had no effect on the local virus titers and the histopathology. Combined treatment with ganciclovir and HIS, however, effectively abolished CMV induced ILD. PMID- 8721548 TI - Overview: issues concerning the pharmacology of multiple drug regimens. PMID- 8721549 TI - Unique clinical trial design: combination acyclovir plus prednisone therapy of localized zoster in the normal host. PMID- 8721550 TI - Design of clinical trials for drug combinations: cytomegalovirus retinitis- foscarnet and ganciclovir. The CMV retinitis retreatment trial. AB - The Cytomegalovirus Retinitis Retreatment Trial was a multicenter clinical trial designed to evaluate three treatments for the treatment of relapsed CMV retinitis; (1) foscarnet; (2) 'high-dose' ganciclovir, and (3) combination foscarnet and ganciclovir. Two hundred seventy-nine patients were enrolled and randomly assigned to one of these three regimens. Patients were followed monthly for 6 months and every 3 months thereafter. Outcomes of interest included: (1) mortality; (2) retinitis progression; (3) change in retinal area affected by CMV; (4) loss of visual field; (5) loss of visual acuity; (6) quality of life; and (7) treatment side effects. PMID- 8721551 TI - CMV retinitis: ganciclovir/monoclonal antibody. PMID- 8721552 TI - Summary: antiviral treatment of hepatitis C virus. PMID- 8721553 TI - Cartesian coordinate analysis of viral burden and CD4+ cell count in HIV disease: implications for clinical trial design and analysis. AB - HIV-1 infection represents a dynamic interaction between the rapid turnover of virus, CD4+ cell proliferation and clearance. HIV-1 disease progression is assessed, in part, by the inverse relationship between virus burden and CD4+ cell count. However, there is enormous individual subject variability between virus burden in the peripheral blood and CD4+ cell count with subsequent disease progression, suggesting that there must be virologic and immunologic modifiers of the inverse relationship between virus load and CD4+ cell count. To investigate these modifiers, we have used a Cartesian coordinate plot analysis to describe the inverse relationship between viral burden and the peripheral blood CD4+ cell count. Subjects from several clinical studies with CD4+ cell counts ranging from < 50 to > 600 cells/microL and varying viral burdens were studied. The analysis described the effect of various virologic and immunologic modifiers on this inverse relationship, for example, viral resistance, viral phenotype and the effect of very low CD4+ cell counts, and specifically addressed individual subject variation in assessing the association between the viral and immunologic parameters that define disease progression and response to antiretroviral therapy. As such, the Cartesian coordinate plot analysis method provides one approach to investigating the individual subject response to antiretroviral therapy. PMID- 8721554 TI - Combination therapy in primary HIV infection. PMID- 8721555 TI - Saquinavir/zidovudine combination in patients with advanced HIV infection and no prior antiretroviral therapy: CD4+ lymphocyte/plasma RNA changes, and emergence of HIV strains with reduced phenotypic sensitivity. PMID- 8721556 TI - Reduced sensitivity to saquinavir: an update on genotyping from phase I/II trials. AB - A genotypic analysis of the HIV-1 proteinase was performed on clinical specimen obtained from patients after different periods of Saquinavir (SQV) treatment. Proteinase genes of integrated proviral DNA from PBMC were isolated by PCR, cloned and individual sequences were obtained. Genotypic resistance was defined by the Gly48-->Val and Leu90-->Met exchanges. Frequencies and kinetics of resistance development will be reported for phase I/II trials V13330. V13329, O13328 and ACTG229 in patients on monotherapy or combination therapy with RT inhibitors. Data from V13330 have been analysed in more detail for correspondence of genotypic and phenotypic resistance and any correlation between resistance and changes in plasma viral RNA load. Furthermore, we will discuss the data from our extensive proteinase gene sequence collection with respect to mutational changes which would be indicative of resistance to other inhibitors of HIV-1 proteinase. PMID- 8721557 TI - Combination therapy with zidovudine, didanosine and saquinavir. AB - The goal of AIDS Clinical Trials Group (ACTG) Protocol 229 was to evaluate the shorter term (24 week) and longer term (48 week) safety and activity of the HIV-1 proteinase inhibitor saquinavir in combination with zalcitabine (ddC) and zidovudine (ZDV) versus saquinavir in combination with ZDV versus ddC in combination with ZDV. This study confirms and extends the conclusions from the first 24 weeks of this study and showed that in subjects with extensive prior antiretroviral experience, the triple combination of saquinavir, ddC and ZDV was well-tolerated, safe and remained superior to the other double drug regimens as measured by CD4+ cell counts, quantitative HIV-1 microculture and plasma HIV-1 RNA levels. PMID- 8721558 TI - D4T nucleoside combinations for HIV. PMID- 8721559 TI - Consensus symposium on combined antiviral therapy; overview of interferon and IL 2 combinations for the treatment of HIV infection. AB - Among the immunomodulatory cytokines that have been evaluated for the treatment of HIV disease, alpha-interferon and interleukin-2 (IL-2) have been the most extensively studied. Monotherapy with alpha-interferon is effective therapy for HIV-associated Kaposi's sarcoma (KS) in patients with CD4 counts > 150 cells/mm3. However, the doses necessary to achieve a significant anti-tumor effect are often poorly tolerated. Combination therapy with alpha-interferon and zidovudine is associated with dose-limiting toxicities and an anti-tumor effect similar to that of higher dose alpha-interferon monotherapy. The combination of alpha-interferon and zidovudine can synergistically inhibit HIV replication in vitro; however, in vivo results suggest the anti-HIV effect of this combination is no greater than that seen with zidovudine monotherapy. Whether combination of interferon-alpha and other antiviral drugs will be useful in the treatment of HIV infection remains to be seen. Recent studies employing intermittent courses of IL-2 combined with continuous antiretroviral therapy indicate that sustained rises in CD4 counts can be achieved. The ability of IL-2 therapy to result in a sustained rise in CD4 counts is critically dependent on the pre-treatment CD4 count. The immunologic and clinical significance of these IL-2-induced increases in CD4 counts is unknown. Larger, controlled trials are currently underway to evaluate the role of intermittent IL-2 therapy in HIV infection. PMID- 8721561 TI - Innovative combination clinical trial designs/patient compliance: a community perspective. PMID- 8721560 TI - Pilot clinical trial of the combination of hydroxyurea and didanosine in HIV-1 infected individuals. PMID- 8721562 TI - Inter-company collaboration for AIDS Drug Development: perspective on combination studies. PMID- 8721563 TI - A new form of skeletal dysplasia with amelogenesis imperfecta and platyspondyly. AB - We report two patients, born of consanguineous parents, affected by a disorder resulting in mild growth retardation. Hallmarks are amelogenesis imperfecta (absence of the enamel cap) associated with brachyolmia-like anomalies: platyspondyly with short pedicles, narrow intervertebral and interpedicular distances, rectangular-shaped vertebrae with posterior scalloping and herniation of the nuclei, and broad femoral necks. Inheritance appears to be autosomal recessive. PMID- 8721564 TI - Alopecia-mental retardation syndrome associated with convulsions and hypergonadotropic hypogonadism. AB - We report two brothers with congenital total alopecia, mental retardation, childhood convulsions and hypergonadotropic hypogonadism. This association has not previously been reported and probably represents a new autosomal recessive condition. PMID- 8721565 TI - Homozygosity and heterozygosity for the transthyretin Leu64 mutation: clinical, biochemical and molecular findings. AB - Transthyretin gene point mutations cause hereditary amyloidosis with an autosomal dominant pattern of inheritance. The disease usually manifests itself in heterozygous patients, although a few homozygotes have been reported. We describe two unrelated patients carrying the Leu64 mutation, one of whom presents a homozygous genotype (Family B). Homozygosity was confirmed by sequence analysis, RG-PCR and double one-dimensional electrophoresis of the plasma protein. Although the clinical picture of the homozygous patient of Family B was more severe than that shown by the heterozygous members of Family A, the variability often displayed by FAP patients does not allow any firm conclusion about the role of homozygosity in the seriousness of the disease. PMID- 8721566 TI - Peripheral blood cell counts in infants with Down's syndrome. AB - The current literature indicates that several abnormalities have been observed in the three hematopoietic cell lines of infants with Down's syndrome. This prospective, longitudinal study was designed to clarify the physiological variation in peripheral blood cell values of children with Down's syndrome by following 25 such infants during their first year of life. Apart from polycythemia in the first week of life, the hemoglobin concentration was, in general, the same as in normal term infants. At 9-12 months of age values for mean corpuscular hemoglobin and mean corpuscular volume tended to be elevated. Serum erythropoietin concentrations were low to normal. White blood cell counts were slightly lower in children with Down's syndrome than in normal children. The study infants had profound thrombocytosis from the age of 6 weeks to the end of follow-up at 1 year. This study, the first longitudinal follow-up of such subjects, indicates that infants with Down's syndrome often have evidence of polycythemia soon after birth and red blood cell macrocytosis and thrombocytosis later in infancy. In conclusion, we carried out peripheral blood cell counts in 25 infants with Down's syndrome, but with no actual hematological disturbance, during their first year of life, and compared them with values for normal term infants. PMID- 8721567 TI - Down syndrome with partial duplication and del (21) syndrome: study protocol and call for collaboration. Study I: Clinical assessment. AB - We report on the clinical and cytogenetic assessment of five cases of Down syndrome phenotype with either a partial duplication of chromosome 21 or a normal karyotype, and we quote a case of del (21q) syndrome. Down syndromes with a partial duplication of chromosome 21 (as well as cases of del (21q), which are partly the phenotypic countertype of trisomy 21) are of paramount importance in the understanding of genes involved in the phenotype of Down syndrome. The goal is to find the relevant genes implicated in the main traits of Down syndrome (i.e. mental retardation, Alzheimer disease, and serious visceral malformations). Such a goal, in our opinion, cannot be reached just by publishing the genotype and the phenotype of a small cohort of patients: 1. a sufficient number of accurate cases is needed, and 2. data have to be computerized for definite conclusions to be reached. The main aims of this report are to present our study protocol and to invite colleagues to participate in a collaborative study in order to collect a maximum of these (rare) cases. PMID- 8721568 TI - The spectrum of ocular features in the Williams-Beuren syndrome. AB - One hundred and fifty-two patients with the Williams-Beuren syndrome were examined to assess the frequency and severity of ophthalmological features associated with the disorder. Eighty-two (54%) had strabismus, all but three, esotropia. One hundred and seventeen (77%) patients had blue irides, 10 (7%) green, and 25 (16%) brown. One hundred and twelve (74%) showed a typical so called stellate iris pattern of the anterior stroma. Whitish anomalies were also detectable in brown irides. Two 9-year-old patients and one 46-year-old patient had initial cataract. Of all the patients with funduscopy, 22% had retinal vascular tortuosity. One patient had suspected Rieger syndrome. Two patients had ptosis, one with a Marcus-Gunn phenomenon. No ocular manifestation of hypercalcaemia was noted. PMID- 8721569 TI - An improved method for the detection of Down's syndrome aneuploidy in uncultured amniocytes. AB - We report a modified method for the rapid detection of aneuploidies directly on human uncultured amniocytes that simplifies and shortens the entire experimental procedure, yielding signals which allow correct diagnosis of trisomy 21 in 97% of cases. The improvement is based on two points: 1) use of cosmid pockets specific for the Down's syndrome minimal region as FISH probes, and 2) a modified protocol for the fixation and preparation of amniocytes. PMID- 8721570 TI - The phenotype of a 45,X male with a Y/18 translocation. AB - In this report, we describe a male infant with a 45,X karyotype; the entire short arm and the centromere of the Y chromosome were translocated onto the short arm of chromosome 18, resulting in an unbalanced dicentric chromosome. Breakpoints were identified by in situ fluorescence hybridization (FISH) on the proximal Yq11 and 18p11.2. Both Y and 18 centromeric alphoid sequences were identified on the derived 18 chromosome. Clinical features were compatible with 18p- syndrome and no Turner stigmata were present in our propositus. Short stature was likely to be related to the deletion of 18p and/or Yq, where a gene involved in stature determination has been located proximal to a gene involved in spermatogenesis (AZF). PMID- 8721571 TI - Two supernumerary marker chromosomes, derived from chromosome 6 and 9, in a boy with mild developmental delay. AB - We report on a boy with two supernumerary marker chromosomes which were identified by fluorescence in situ hybridization and derived from chromosome 6 and 9. In lymphocytes, a mosaic karyotype was found: 46,XY (17%)/ 47,XY,r(6) (24%)/47,XY,r(9) (20%)/48,XY,r(6),r(9) (39%). Only minor dysmorphic features and mild developmental delay were present. Despite extensive fluorescence in situ hybridization studies using a large panel of probes, we were unable to characterize the marker chromosomes in more detail, mainly because no probes for the chromosome regions involved were available to us. In order to reach a better understanding of the clinical relevance of small supernumerary marker chromosomes, it will be necessary to create a widely available set of probes, covering all chromosome regions. PMID- 8721572 TI - Generalized osteoporosis in a patient with oculocutaneous hypopigmentation syndrome (OOCHS), without cerebral defects. A new syndrome? AB - An 8-year-old male patient presented a unique pattern of congenital anomalies. Prominent findings included a combination of severe osteoporosis and congenital oculocutaneous hypopigmentation. The patient may represent a hitherto undescribed syndrome of unknown etiology. PMID- 8721573 TI - Supernumerary ring chromosome 20 characterized by fluorescence in situ hybridization. AB - We report on a boy with mild dysmorphic features and developmental delay, in whom karyotyping showed an additional minute ring chromosome in 60% of metaphases. Fluorescence in situ hybridization (FISH) with a centromere specific probe demonstrated that the ring chromosome contained the centromeric region of chromosome 20. The ring was highlighted completely using a chromosome 20 painting probe. A cosmid probe for 20p 12-13 gave a positive signal and hybridization with an all-telomere probe showed on signal, suggesting a breakpoint in the 20p telomere. The results suggested that only a small part of 20q was involved in this ring. The ring was also detected in 18% of nuclei of a buccal smear. The phenotypic similarities of symptoms in the proband to patients with a (partial) trisomy 20p and the dissimilarities to symptoms in patients with (partial) trisomy 20q were in agreement with the FISH results. PMID- 8721574 TI - G542X mutation in Mexican cystic fibrosis patients. AB - We analyzed the frequency of the G542X mutation in a sample of 76 Mexican cystic fibrosis patients and the genotype-phenotype correlation. The mutation was screened using PCR-mediated site-directed mutagenesis, and was present on 7.2% of the CF chromosomes. This frequency is significantly higher than the worldwide frequency according to the Cystic Fibrosis Genetic Analysis Consortium (3.4%, p < 0.01), and similar to that reported in Spain (8%), which is in accordance with the ethnic origin of the Mexican population. All patients carrying G542X on at least one allele had mild to moderate pulmonary disease. In patients with hepatobiliary involvement, the frequency of G542X chromosomes was higher than the frequency of the mutation in all the Mexican CF chromosomes. PMID- 8721575 TI - [Clinical value of positron emission tomography (PET) in oncologic questions: results of an interdisciplinary consensus conference. Schirmerreschaft der Deutschen Gesellschaft for Nuklearmedizin]. AB - AIM: The purpose of the present paper is to assess clinical value of PET in oncology on the basis of published studies. METHODS: Clinical value of PET in oncology was evaluated by a panel of recognized experts in the framework of an interdisciplinary consensus conference. On the basis of PET studies, well documented in the international literature, the value of PET for solving clinical questions was classified according to the following categories (classes 1a, 1b, 2a, 2b, 3): "appropriate" (1a), "mostly acceptable" (1b), "helpful" (2a), "value as yet unknown" (2b), "useless" (3). RESULTS: 2-fluorodeoxyglucose (FDG) acts as the radiopharmaceutical of choice for PET in clinical oncology. PET is indicated (1a) for diagnosing relapse in high grade glioma (FDG) or low grade glioma (C-11 methionine or F-18 fluorotyrosine), differential diagnosis of solitary peripheral pulmonary nodules in high risk patients and for diagnosis of pancreatic carcinoma. PET may be clinically used (1b): In "low-grade" glioma, search for unknown primary in head and neck tumors, suspicion of relapse in non-small cell bronchial carcinoma (NSCBC) and colorectal carcinoma, lymphnode staging in NSCBC, pancreatic carcinoma, muscle invasive bladder carcinoma and testicular cancer. Staging of Hodgkin's disease (HD, stage I/II vs III), early therapy control in patients with a residual mass or suspicion of relapse in HD and in high grade NHL, lymph node staging and search for distant metastases in malignant melanoma (Breslow > 1.5 mm), search for lymph node or distant metastases in differentiated thyroid cancer with elevated hTG and a negative radioiodide whole body scan. Many further indications are emerging, but are not yet sufficiently well documented in the literature. For most indications beside scientific studies, an individual cost benefit utility evaluation by the responsible physician is recommended. CONCLUSION: Metabolic imaging of PET provides for many principle advantages compared to conventional anatomically based cross sectional imaging. For routine use in oncology a detailed assessment of specific efficiency of PET is indicated. PMID- 8721576 TI - [111IN-octreotide and 123I-MIBG scintigraphy in the diagnosis of small intestinal carcinoid tumors--results of a comparative investigation]. AB - AIM: This study compares the diagnostic value of 111In-Octreotide- and 123I-MIBG scintigraphy in the diagnostic imaging of carcinoids and correlates the results with CT findings. METHODS: Twenty-eight patients underwent octreotide-, MIBG scintigraphy and CT examination within a short time period. RESULTS: In 13/15 preoperatively examined patients somatostatin receptor expression of the tumor was demonstrated; MIBG-uptake was found in 8/15 cases. Octreotide and MIBG scintigraphy revealed liver metastases in 26 and 19 cases, respectively. CT examinations detected liver metastases in 22 cases. The number of metastatic lesions found by MIBG and CT was lower than with octreotide. In 3 patients unknown mediastinal lesions were detected by the octreotide scan, 2/3 did not show MIBG uptake. In 5 non-hormone secreting tumors shown by octreotide scintigraphy, no MIBG uptake was found. With respect to the diagnostic accuracy octreotide scintigraphy appeared superior to MIBG. CONCLUSION: MIBG scans do not provide additional diagnostic information compared to octreotide scintigraphy. MIBG scintigraphy may be potentially useful in terms of planning 131I-MIBG therapy only. PMID- 8721577 TI - [Validity of bone scintigraphy for radiotherapy treatment planning of the skeletal system]. AB - Aim of this paper is to assess the validity of bone scanning for radiation therapy treatment planning in the skeletal system. Radiation therapy treatment planning in the skeletal system is based on plain film radiographs and bone scanning. Between January 1, 1993 and June 30, 1994, 228 patients were irradiated due to malignancies in the skeletal system. Included were bone metastases, lymphomas, sarcomas and bone invasion of neuroblastoma. The morphological display of the plain film radiographs and the bone scintigraphies were compared and modifications of the target volume due to the scan findings quantified. In 17 of 228 patients (7.5%) the target volume was modified by at least 2 cm due to tumor invasion depicted by radionuclide scanning only. Bone scintigraphy enables a sensitive display of bony invasion in skeletal malignancies. Because of a scan induced alteration of the target volume in 7.5% of the cases, bone scanning cannot be abandoned in RT treatment planning. PMID- 8721579 TI - [Molecular biology of thyroid diseases]. AB - Recent developments in molecular biology and the accessibility of techniques for clinical research have led to a better understanding of the background of common thyroid diseases. The cloning and sequencing of the thyroid stimulating hormone receptor, thyroid peroxidase and thyroglobulin, and the characterization of the protein-DNA interaction during thyroid hormone action, as well as the discovery of intracellular signal transduction pathways were the most important steps which resulted in new diagnostic and therapeutic approaches. New explanations of thyroid autoimmune processes are being investigated. PMID- 8721578 TI - [Pain therapy with rhenium-186 HEDP in multiple bone metastases]. AB - AIM: Therapeutic means for patients with painful bony metastases are strongly limited. In these patients, a systemic therapy with isotopes can be helpful. The aim of this study was to evaluate the efficacy of rhenium-186 HEDP for pain palliation in patients with disseminated bone metastases. METHODS: 30 patients taking analgesics because of bone pain received one or more injection of 1295 MBq 186Re HEDP for pain palliation. RESULTS: In 70% of the patients, therapy resulted in a significant reduction of pain. The average duration of pain relief was 4 weeks (1 week-2.5 months). The main side effects of therapy were a decrease of platelets (aver. 30000/microliters) and an increase of pain for 1-2 days (flare phenomenon). CONCLUSION: Therapy with rhenium-186 HEDP can be used complementarily to analgesic therapy in patients with painful, disseminated bone metastases. PMID- 8721580 TI - [Endoscopic management of bile leakage following laparoscopic cholecystectomy]. AB - During a 15-month period 8 patients (1 male, 7 females, mean age 49 years) admitted from different institutions were treated with bile leaks after laparoscopic cholecystectomy. The diagnosis was established by endoscopic retrograde cholangiography. The site of the leakage was the cystic duct in five cases, the common bile duct in one case, the hepatic "bed" in one case and it was undetectable in one case. Five patients had common bile duct stone and one patients also had common bile duct stricture. Endoscopic sphincterotomy was performed in all cases with stone extraction if needed, and stent insertion (7 or 10 Fr straight endoprosthesis) in six cases. The stents were removed after 4-16 weeks in all cases, except the patients with common bile duct stricture, in whom the stent was exchanged for two other endoprostheses to prevent restricture after three months, she is still under treatment. Endoscopic sphincterotomy, stone removal and stent placement were successful in all cases. Symptoms resolved in hours, and the bile leakage stopped in days. Apart from transient serum amylase level elevation in one patient no other complication was observed. CONCLUSION: The endoscopic treatment of bile leakage after laparoscopic cholecystectomy has proved to be a safe and effective method. It may be considered the method of choice in the management of this complication. PMID- 8721581 TI - [Intermittent Romano-Ward syndrome]. AB - Two cases are described to have intermittent long QT syndrome. This type of long QT syndrome is not easy to recognize and the result could be fatal in case taking I/A type antiarrhythmic drugs. The first patient who was investigated had got "chinidin syncope" two years earlier, the second one had symptoms of syncope and dizziness for 3 months. The long QT syndrome was diagnosed in both cases by Holter monitoring. For these reason this paper underline the value of Holter monitoring as a diagnostic tool to establish the diagnosis of long QT syndrome. The monophasic action potential showed early after depolarisations in both cases. PMID- 8721583 TI - [Genetic counseling in a case of Takayasu arteritis]. AB - Two cases affected with Takayashu arteritis are presented. The adult probanda born in 1960 after genetic counselling and appropriate prenatal care had a healthy liveborn boy. Her previous six pregnancies were terminated on the basis of forced medical reason. The infant probanda died at the age of 4 months with the features of sudden infant death syndrome (SIDS). Pathohistological examination detected her Takayashu arteritis. It seems to be the youngest published case in the international literature and Takayashu arteritis may be the rare cause of SIDS. PMID- 8721582 TI - [The role of Fc gamma R specific autoantibodies in autoimmune diseases. Review of the literature and authors' own research]. AB - The authors characterize the possible role of circulating autoantibodies specific for Fc gamma receptors in autoimmune diseases. Dysfunction of Fc gamma receptors, resulting in prolonged clearance of immune complexes, has been observed in both systemic and organ specific autoimmune disorders. Although the mechanism of the defective Fc gamma receptor function is not clearly understood, some findings suggest that Fc gamma receptor specific autoantibodies may have significant part in these processes. The authors found high titers of anti-Fc gamma receptor autoantibodies in the sera of several mouse strains prone to autoimmune diseases, and then extended these observations to human autoimmune diseases. They analized the specificity and functional effects of these antibodies. The authors hypothesize, that Fc gamma receptor specific autoantibodies may play a role in the pathology of autoimmune diseases by stimulating Fc gamma-receptor-bearing cells to release oxygen intermediates, different lysosomal hydrolases and cytokines or by blocking the phagocytosis of immune complexes. PMID- 8721584 TI - Eccrine or apocrine poroma? Six poromas with divergent adnexal differentiation. AB - We describe six cases of benign eccrine poroma-like neoplasms with divergent adnexal differentiation. Four cases exhibited sebaceous differentiation in the form of individual or clustered sebocytes with or without sebaceous ducts. One case showed both sebaceous and hair follicle differentiation, and one case showed sebaceous and possible apocrine secretory differentiation. Clinically, most were skin-colored, red, or purple papules or nodules. One patient had a preoperative diagnosis of Bowen's disease, with an erythematous plaque. None recurred following biopsy. Previous reports of similar lesions have suggested a possible role for human papilloma virus (HPV) in their pathogenesis; however, immunohistochemical staining for HPV structural antigens was negative in all six of these cases. Similarities to previously reported cases of eccrine poroma-like neoplasms with sebaceous differentiation are discussed. Given the evidence of sebaceous and follicular differentiation seen in this study and the common embryologic origin of follicular, sebaceous, and apocrine structures, it follows that at least some benign neoplastic proliferations with histopathologic features of "eccrine" poroma could be of apocrine origin. PMID- 8721585 TI - Dermatoscopic and videomicroscopic features of melanocytic plantar nevi. AB - Nearly 10% of Japanese people have pigmented nevi on the soles. Since malignant melanoma also occurs on the plantar area in the Japanese, it would be very valuable to be able to differentiate benign and malignant lesions in the early clinical state. We have investigated the epiluminescence microscopic features of 500 melanocytic nevi on the soles of Japanese people using a dermatoscope and a videomicroscope that can magnify lesions from x 10 to x 200. The results showed that the surface profile of benign melanocytic nevi is mainly classified into five types; that 9% of plantar nevi, however, do not fit into this classification and are categorized as a miscellaneous type; and that the other nonmelanocytic disorders, such as verruca vulgaris and black heel, are easily differentiated by their surface profile. More important, the histological examination showed that atypical nevi, malignant melanoma in situ, and acral lentiginous melanoma are exclusively compartmentalized in the miscellaneous type of surface profile. Our data suggested that epiluminescence microscopy may be a useful method for discrimination of plantar benign and malignant melanocytic lesions. PMID- 8721586 TI - Atypical cutaneous changes after topical treatment with nitrogen mustard in patients with mycosis fungoides. AB - Side effects in the treatment of mycosis fungoides with topical nitrogen mustard include allergic contact dermatitis, hyperpigmentation, urticaria, and erythema multiforme-like dermatitis. We reviewed biopsy specimens from 10 patients with mycosis fungoides who were treated with topical nitrogen mustard for 10-76 months. There was no history of oral psoralen with long-wave UV radiation treatment, radiotherapy, or systemic chemotherapy. Control biopsies taken from erythematous or poikilodermatous patches on the trunk or proximal extremities showed epidermal and dermal changes associated with cytologic atypia that were not present before treatment. These changes included slight epidermal hyperplasia with foci of flat rete ridges, atypical keratinocytes with large nuclei, mostly in the lower portion of the epidermis; suprabasal mitotic figures; a few dyskeratotic cells, focal vacuolar alteration of the epidermal basal layer; increased number of slightly enlarged junctional melanocytes; melanophages in the papillary dermis; dilated blood vessels lined by plump, atypical endothelial cells; and large fibroblasts with atypical nuclei. These atypical histologic changes resemble, in part, those described in association with systemic chemotherapeutic agents, such as etoposide, busulfan, and bleomycin. We conclude that topical nitrogen mustard should be added to the list of chemotherapeutic agents that can produce atypical histologic changes in the skin. PMID- 8721587 TI - Caterpillar bodies of porphyria cutanea tarda ultrastructurally represent a unique arrangement of colloid and basement membrane bodies. AB - Caterpillar bodies are eosinophilic, periodic acid-Schiff (PAS)-positive globules arranged in a linear fashion in the epidermis overlying subepidermal blisters of porphyria cutanea tarda (Am J Dermatopathol 1993;15:199-202). We retrospectively studied by transmission electron microscopy nine cases of porphyria cutanea tarda (PCT) that demonstrated caterpillar bodies. We identified three components of the eosinophilic bodies: degenerating keratinocytes, colloid bodies, and basement membrane bodies. The colloid bodies consisted of whorled masses of filaments containing degenerating melanosomes, vacuoles, mitochondria, and desmosomes. Basement membrane bodies were composed of convoluted basement membrane material and associated collagen. Both colloid and basement membrane bodies were often associated with degenerating keratinocytes, were located both intra-and extracellularly, and were occasionally fused to one another. We believe that caterpillar bodies are a combination of degenerating keratinocytes, colloid bodies, and basement membrane bodies formed by repeated blistering and reepithelialization with transepidermal migration. Furthermore, we believe that caterpillar bodies are a diagnostic clue for the diagnosis of PCT. PMID- 8721588 TI - Cutaneous pathology in primary erythermalgia. AB - Primary or idiopathic erythermalgia is characterized by recurrent, red, warm, and painful lower extremities. It arises at young age and persists throughout life because no treatment is available. We report the cutaneous pathology of affected skin lesions of three patients with primary erythermalgia. Biopsy specimens showed a mild perivascular mononuclear infiltrate, thickened blood vessel basement membranes, abundant perivascular edema, and moderate endothelial swelling. The thickened basal membrane of the blood vessels showed a laminar structure, and abundant perivascular edema and moderate endothelial cell swelling were evident. These histopathologic findings in primary erythermalgia appear to be nonspecific but allow diagnostic differentiation from erythromelalgia in which fibromuscular intimal proliferation and occlusive thrombi in the endarteriolar capillaries are apparent and from erythermalgia secondary to vasculitis. Histopathologic examination of affected skin lesions in patients with red, congested, warm, and painful burning extremities is a valuable tool in the diagnostic process. PMID- 8721589 TI - Immunohistochemical study of primary and recurrent basal cell and metatypical carcinomas of the skin. AB - We investigated cell proliferation and expression of cytoskeletal proteins in 32 cases of primary basal cell carcinomas (BCC), 10 cases of recurrent BCC, and 10 cases of metatypical carcinomas (MTC). Paraffin-embedded biopsies were evaluated immunohistochemically with a battery of antibodies. Antibodies to proliferating cell nuclear antigen (PCNA) demonstrated comparatively low numbers of proliferating cells in 25 of 32 cases of primary BCC. In contrast, both recurrent BCC and MTC exhibited three to four times higher levels of proliferating cells than primary BCC. PCNA-positive cells were usually distributed uniformly throughout the lobules; at times, however, they were localized to the outer areas of those neoplasms, with a comparatively low level of proliferation index. Antibodies to keratin 17 strongly stained cells of all BCC cases, and antibodies to keratin 8 reacted with most of them. In contrast, the staining intensity of both types of keratin in MTC was decreased six to eight times as compared with all BCC. In addition, cells of eight BCC and three MTC reacted with antibodies to smooth muscle alpha-actin and myosin, neoplasms that did not differ by the number of PCNA-positive nuclei from carcinomas without contractile proteins. The differences in cell proliferation and keratin expression between BCC and MTC may be useful criteria for further distinguishing these carcinomas. The appearance of contractile proteins in some BCC and MTC may be the result of, or implies, myoepithelial differentiation. PMID- 8721590 TI - Basal cell carcinoma arising in association with desmoplastic trichilemmoma. AB - We encountered three cases of basal cell carcinoma (BCC) and one case of an atypical basaloid proliferation best classified as a BCC arising in association with desmoplastic trichilemmomata. Desmoplastic trichilemmoma is a benign tumoral lesion with a pseudomalignant appearance that combines histomorphological features of an infiltrating neoplasm with those of a benign trichilemmoma. PMID- 8721591 TI - Free eosinophil granules in urticaria: a correlation with the duration of wheals. AB - To determine the factors associated with the duration of wheals in urticaria, we carried out a quantitative study of mast cells and eosinophils in both peripheral bloods and skin lesions. The percentage of cytoplasmic vacuolation, indicating a result of eosinophil degranulation of all types of specific granules of circulating eosinophils was significantly increased in the long-duration group compared with the short-duration group. Many extracellular free eosinophil granules were observed in the vicinity of disrupted eosinophils in the long duration wheals. On the other hand, the short-duration wheals had few free eosinophil granules. The number of free eosinophil granules and the percentage of free eosinophil granules to eosinophils were significantly different between the long-and the short-duration wheals, whereas there was no significant difference between the early stage and the late stage of the long-duration wheals. The numbers of all inflammatory cells, eosinophils, and activated mast cells were increased in the late stage of the long-duration wheals. These findings suggest that free eosinophil granules liberated from infiltrating eosinophils by some stimulus may provoke the persistent activation of mast cells, and the mast-cell derived mediators may induce the persistent state of inflammation in the long duration wheals. PMID- 8721592 TI - Folliculosebaceous cystic hamartoma with perifollicular mucinosis. AB - An 8-year-old boy had a congenital, solitary verrucous lesion on his ear that histologically showed numerous maloriented infundibulocystic structures from which radiated immature sebaceous lobules as well as hair germs with rudimentary papillae. The surrounding stroma was fibrillary, containing large quantities of mucin. We conclude that perifollicular mucinosis, considered to be specific to Carney's syndrome, can also be observed in solitary folliculosebaceous hamartoma. PMID- 8721593 TI - Reactive angioendotheliomatosis. Case report and review of the literature. AB - Angioendotheliomatosis is an uncommon disease characterized histologically by proliferation of cells within vascular lumina with secondary intravascular thrombi resulting in obliteration of the involved vessels. While angioendotheliomatosis was initially thought to be a single disease entity, recent studies show that the disease may be divided into benign (reactive) and malignant variants, with the malignant variant representing intravascular malignant lymphoma. The reactive variant is rare, with only 16 cases reported in the literature, and is characterized histologically by proliferating endothelial cells within vessel lumina. In this report we present a case of reactive angioendotheliomatosis and review the literature on this rare entity. PMID- 8721594 TI - Cutaneous sparganosis. AB - Sparganosis is an infection caused by migrating larvae of the cestode genus Spirometra. There have been approximately 62 cases of sparganosis reported in the United States. Although a subcutaneous mass is the most common manifestation, sparganosis is not well-described in the dermatology literature. We present a case of cutaneous sparganosis in a 52-year-old Filipino American woman. Histologically, the sections showed a granulomatous panniculitis and dermatitis containing a section of a sparganum. A transverse section of an intact sparganum reveals an eosinophilic cuticle, loose stroma, calcareous bodies, and smooth muscle fibers. PMID- 8721595 TI - Adenoma of anogenital mammary-like glands. AB - The authors describe a benign adenomatous neoplasm arising from anogenital mammary-like glands. These glands have been recently recognized in the anogenital region and may play an important role in the development of some neoplastic disorders peculiar to this anatomical area. Important features of the tumor are adenomatous tubular and cystic structures with apocrine-like cytologic features, a lobular configuration, and abundant fibrous stroma. PMID- 8721596 TI - Cutaneous adnexal carcinoma arising within a solitary cylindroma-spiradenoma. AB - Malignant transformation within an adnexal neoplasm with features of spiradenoma and cylindroma is reported in a 60-year-old woman. The malignant component of the tumor consisted of areas of solid carcinoma, with focal squamous metaplasia. In the benign component, apart from the features of spiradenoma and cylindroma, sebaceous differentiation was present adjacent to a cystic space lined by apocrine epithelium. The morphologic composition of this unusual neoplasm supports the concept of a histogenetic relationship between cylindromas and spiradenomas, including their differentiation toward the pilosebaceous-apocrine unit. PMID- 8721597 TI - Malignant chondroid syringoma: immunohistopathology. AB - Malignant chondroid syringoma, also called malignant mixed tumor of the skin, is a rare variant of a malignant tumor derived from sweat gland cells. Histologically the tumor is composed of an epithelial and a mesenchymal component. The epithelial structures show glandular differentiation and carcinomatous features. They are embedded in a mucinous stroma with spindle mesenchymal cells and areas of chondroid differentiation. The epithelial cells express cytokeratins. The luminal epithelial cells show binding to the lectin Ulex europaeus; intraluminal cells are carcinoembryonic antigen positive. The stromal cells are cytokeratin negative and sporadically positive for vimentin. Chondroid areas are S-100 protein and vimentin positive. No labeling for actin is seen. PMID- 8721598 TI - Hair follicle nevus: case report and review. AB - Hair follicle nevus has been variously defined as a specific type of neoplasm with follicular differentiation or as a synonym for trichofolliculoma; the latter view has predominated in recent years. We report a nodular lesion that was identified histopathologically as a hamartoma with follicular differentiation and that systematic serial sectioning showed not to be a trichofolliculoma. After reviewing previous descriptions of lesions diagnosed as hair follicle nevus, and in the light of our own observations, we believe that hair follicle nevus is a distinct type of adnexal hamartoma. PMID- 8721599 TI - Metastatic endometrial adenocarcinoma to the skin of a toe. AB - A 55-year-old woman had a history of prolonged vaginal bleeding. Dilatation and curettage confirmed poorly differentiated endometrial adenocarcinoma. After hysterectomy, salpingo-oophorectomy, and 5,040 cGy of postoperative irradiation, lung metastases developed, which temporarily responded to chemotherapy that included adriamycin and cisplatin. Metastatic lesions developed later in the left acetabulum and the hallux. The latter, clinically reminiscent of pyogenic granuloma, affected only skin. The poorly differentiated toe metastasis duplicated areas of the primary endometrial adenocarcinoma and was immunohistochemically similar to it. While appearing quite rarely, and generally late in the disease, pedal skin lesions may represent metastases from known or occult visceral cancers. PMID- 8721600 TI - Gleanings from the attic: medical references in the Noctes Atticae of Aulus Gellius. PMID- 8721601 TI - Annular lesions in exogenous ochronosis. PMID- 8721602 TI - Well-differentiated malignant cylindroma. PMID- 8721603 TI - Primary cutaneous osteosarcoma. PMID- 8721604 TI - [Distribution of polyamines in Escherichia coli and their role in potassium exchange between the cell and the environment during aerobiosis-anaerobiosis transitions]. AB - The transition of E. coli cells to anaerobiosis is accompanied by the onset of two cellular cation flows in different directions: potassium is released into the environment, and putrescine enters the cells. If aerobic conditions are reestablished, the cation flows change directions. Under anaerobiosis, the cell components bind putrescine. Investigation of the chemical interactions resulting in putrescine binding and metabolic conversion under anaerobic conditions revealed DNA to be the main target in this process. The driving forces and mechanisms of cation transfer are discussed, and the involvement of putrescine in topological changes in the DNA and the development of adaptive cell responses to anaerobiosis is considered. PMID- 8721605 TI - [Accumulation in the medium and properties of proteolytic enzymes formed by Nocardia minima]. AB - The accumulation dynamics of extracellular proteinases with plasmin-like and activatory mechanisms of action (belonging to the subtilisin family of the class of serine proteases and to the class of metallo-proteinases, respectively) were followed during the course of growth of Nocardia minima, strain 1. The sequence of the appearance of extracellular proteinases and their activity at various growth phases of the producer suggest their participation in the processes of the morphophysiological transformations accompanying the transition from mycelium to rods and from rods to cocci. The proteinases produced by Nocardia showed specificity to either human or bovine blood fibrin. The specific pattern of Nocardia minima proteinases (absence of chymotrypsin-like enzymes typical of streptomycetes, presence of metalloproteinases typical of bacilli) and the distinctive features of Nocardia minima subtilisins distinguishing them from subtilisins of bacilli and streptomycetes agree well with the intermediate taxonomic position of N. minima. PMID- 8721606 TI - [Physiological effects of sunlight on Bacillus subtilis strains with varying capacity to repair photoproducts]. AB - Nonlethal doses of sunlight were established to cause a growth delay in Bacillus subtilis strain 1201 possessing an unimpaired repair system. This effect was not observed with the 1201-10 uvr1 mutant deficient in DNA excision repair; however, addition of an extract of uvr1+ cells restorated the phenomenon of growth delay by sunlight in this mutant. Experiments with glass light filters showed that a weakened growth delay in strain 1201 could also be produced by rays of sunlight with wavelengths greater than 380 nm. Maximum growth delay was observed after irradiation with the complete spectrum of sunlight. Experiments with the protein synthesis inhibitor chloramphenicol indicated that sunlight promotes synthesis of proteins preventing the dying of cells. The presence of Casamino acids is favorable for the synthesis of protective proteins. PMID- 8721607 TI - [Bioantimutagenic effect of the culture fluid of Streptococcus faecalis on mutagenesis induced by 2-nitrofluorene in Salmonella typhimurium TA 1538 and TA 98]. AB - Streptococcus faecalis growing in rich organic media or synthetic medium containing 0.1% trypton and 0.05% yeast extract was shown to form substances with a bioantimutagenic activity against the mutagenicity of 2-nitrofluorene and 4 nitroquinoline-1-oxide, which induce frameshift and base-pair substitution mutations, respectively. The maximum accumulation of antimutagens in the medium was observed within 16-24 h of growth. The modified Ames test showed that the culture liquid of Str. faecalis possessed a higher antimutagenic activity for Salmonella typhimurium strain TA 1538 than for strain TA 98 with an enhanced SOS response. Fractionation of the culture liquid revealed that the bioantimutagenic activity is localized in the fraction of peptides with molecular masses of less than 12 kDa. PMID- 8721608 TI - [Dependence of the intensity of 2,4,6-trinitrotoluene-induced mutagenesis on the physiological state of tester strains of Salmonella typhimurium]. AB - The dependence of the mutagenic potential of 2,4,6-trinitrotoluene (TNT) on the growth phase of the Salmonella typhimurium tester strains TA100 and TA98 is shown. The mutagenic potential proved to be maximal for log-phase cultures, being higher with strain TA100 than TA98. The amount of TNT absorbed by bacteria in the first minutes of contact varied, depending on culture age, and was the largest at the beginning of the exponential growth phase. A direct relation between the number of microbial cells and the amount of TNT absorbed from the medium was shown. The analysis of infrared (IR) spectra of cells after a 3-min contact with TNT, demonstrated that, at physiological temperatures, 2.4% of the original amount of TNT was present in cells of the tester strain in an unaltered form, while, at a lowered temperature, this value was 10.3%. Thus, the retardation of physiological processes in the cell changes the intensity of mutagen absorption and transformation. It is emphasized that an accurate assessment of mutagenicity should take into account the physiological state of tester bacteria. PMID- 8721609 TI - Nitric oxide exposure and sulfhydryl modulation alter L-arginine transport in cultured pulmonary artery endothelial cells. AB - The effect of nitric oxide (NO) exposure and sulfhydryl-reactive chemicals on L arginine transport in pulmonary artery endothelial cells was evaluated. Exposure of pulmonary artery endothelial cells to 7.5 ppm (0.4 microM) NO for 4 h resulted in a significant (p < 0.05) reduction of Na(+)-dependent but not Na(+) independent L-arginine transport. More prolonged exposure for 12-24 h reduced both Na(+)-dependent and Na(+)-independent transport of L-arginine with maximal loss of transport after 18 h of exposure (p < 0.02 for both). Similarly, incubation of cells in the presence of 50-200 microM S-nitroso-acetyl penicillamine (SNAP) (but not 500 microM each of nitrate or nitrite) for 2 h also reduced both the Na(+)-dependent and Na(+)-independent transport of L-arginine (p < 0.05 for all concentrations). The SNAP-induced reduction of L-arginine transport was blocked by the NO scavenger oxyhemoglobin. When cell monolayers were exposed to varying concentrations of the sulfhydryl reactive chemicals N ethylmaleimide (NEM) and acrolein, a dose-dependent reduction of L-arginine transport by both Na(+)-dependent and Na(+)-independent processes was observed. Na(+)-dependent L-arginine transport was more susceptible to inhibition by exposure to NO and to sulfhydryl reactive chemicals. Incubation of cells with 0.5 mM of the thiol-containing agent N-acetyl-L-cysteine prior to and during NEM or acrolein exposure blocked NEM and acrolein-induced reduction of L-arginine transport by both Na(+)-dependent and Na(+)-independent processes. Similarly, NO induced reductions of Na(+)-dependent and Na(+)-independent L-arginine transport were reversed to control levels 24 h after termination of NO exposure. Treatment with the disulfide reducing agent dithiothreitol after exposure to NO resulted in partial reversal of the decreases in L-arginine transport. These results demonstrate that exposure to exogenous NO is responsible for reversible reductions of plasma membrane-dependent L-arginine transport mediated by both the Na(+)-dependent (system Bo,+) and the Na(+)-independent (system y+) transport processes. Modulation of the sulfhydryl status of plasma membrane proteins involved in L-arginine transport, such as L-arginine transporters and/or Na+/K(+) ATPase, may be responsible, at least in part, for reductions in overall L arginine transport in pulmonary artery endothelial cells. PMID- 8721610 TI - Lipid peroxidation in neural tissues and fractions from vitamin E-deficient rats. AB - A severe and chronic deficiency of vitamin E results in a characteristic neurological syndrome in both man and experimental animals. This is presumed to result from increased oxidative stress arising from a reduction in antioxidant capacity. In this study we have examined parameters of endogenous lipid peroxidation and susceptibility to in vitro oxidative stress of neural tissues and fractions, and some non-neural tissues from 1-year-old vitamin E-deficient and control rats. We have shown: (1) an increase in endogenous lipid peroxidation (thiobarbituric acid reactive substances and malondialdehyde) in neural tissues from vitamin E-deficient animals compared to controls. (2) The following order of susceptibility of neural tissues to in vitro oxidative stress in both vitamin E deficient and control animals: brain >> muscle > cord > nerve. (3) The susceptibility of different brain regions to in vitro oxidative stress varied in a consistent manner with the cortex, striatum, and cerebellum showing the greatest and brainstem and hypothalamus the least susceptibility. (4) Fractions isolated from myelinated nerves of brainstem showed the following order of susceptibility to in vitro oxidative stress: axoplasmic membranes and organelles > axolemma enriched fraction > whole homogenate > = myelin. These results would fit with the characteristic neuropathology associated with severe and chronic vitamin E deficiency. PMID- 8721612 TI - Mechanism of tetrahydroxy-1,4-quinone cytotoxicity: involvement of CA2+ and H2O2 in the impairment of DNA replication and mitochondrial function. AB - In this work we investigated the toxicity of a polyphenolic p-benzoquinone derivative, the tetrahydroxy-1,4-quinone (THQ) toward V79 Chinese hamster fibroblasts and analyzed the role of H2O2 and Ca2+ in that mechanism. The exposure of exponentially growing cultures to THQ, in the presence of 1.0 mM Ca2+, caused a dose-dependent inhibition of cell growth and DNA synthesis. Complete prevention of those effects by catalase indicated that H2O2-induced damages should underlie both toxic processes. Further detection of a rise in the intracellular free Ca2+ concentration ([Ca2+]i) in cells exposed to THQ plus Ca2+, together with the partial protection conferred by the intracellular Ca(2+) chelator fura-2 against cell growth inhibition, indicated that a disruption of Ca2+ homeostasis is a determinant event in THQ cytotoxicity. Furthermore, the intracellular accumulation of rhodizonic acid (RDZ), the primary oxidative product of THQ, indicated that THQ, or its corresponding semiquinone form, was entering the cells and undergoing further autoxidation to RDZ. It was also evidenced that mitochondria represent an important target in the development of THQ toxicity, as shown by the disruption of the transmembrane electrical potential (delta psi) of isolated rat liver mitochondria, as well as by the Ca(2+)-release by mitochondria of permeabilized V79 cells. We concluded that disruption of Ca2+ homeostasis and generation of H2O2 are critically involved in THQ-induced impairment of DNA replication and mitochondrial functions, leading ultimately to cell growth inhibition. PMID- 8721611 TI - Glucose, glutathione, and cellular response to spermine oxidation products. AB - Bovine serum amineoxidase (BSAO) oxidatively deaminates polyamines, which contain primary amine groups with formation of several toxic products, H2O2, and aldehyde(s). We evaluated the role of glucose metabolism via the pentose phosphate cycle and the level of intracellular glutathione on cytotoxicity induced by each of the toxic products in Chinese hamster ovary (CHO) cells. Glucose protected cells against cytotoxicity in the presence of BSAO at low spermine concentrations ( < 50 microM), where H2O2 was the only toxic species present. When catalase was present, cytotoxicity is attributed to spermine derived aldehyde(s). Glucose did not protect cells against cytotoxicity induced by spermine-derived aldehyde(s), nor by the aldehyde acrolein. Hydrogen peroxide produced by spermine and BSAO stimulated pentose cycle activity, whereas the aldehyde(s) did not. Depletion of intracellular glutathione with L-buthionine sulfoximine (1 mM, 24 h) sensitized cells to the cytotoxic effects of both H2O2 and the aldehyde(s) produced by spermine and BSAO. The pentose cycle and the glutathione redox cycle have an important role in protection against H2O2 generated from spermine oxidation. Glutathione appears to have a role in protecting cells against cytotoxicity attributed to spermine-derived aldehyde(s), most likely by conjugation in a reaction catalyzed by glutathione S-transferase, whereas metabolism of glucose via the pentose cycle did not. The metabolism of both glucose and glutathione, affect the cellular response to H2O2 and aldehyde(s) derived from spermine, although different pathways are involved. PMID- 8721613 TI - Functional alterations in sarcoplasmic reticulum membranes of magnesium-deficient rat skeletal muscle as consequences of free radical-mediated process. AB - Free radical-induced physiopathologies are generally thought to be mediated by membrane injuries. Using a pro-oxidant model induced by dietary magnesium deficiency, we have recently shown that skeletal muscle lesions occurred with a rise in the calcium level and enhanced free radical production. In this study, we investigated the physicochemical and biochemical properties of sarcoplasmic reticulum membranes isolated from hind limb muscles of weanling male rats pair fed magnesium-deficient or control diets for 12 d. The calcium-induced calcium efflux from preloaded vesicles was increased in membranes isolated from Mg deficient rat muscle. In agreement with this latter observation, we demonstrated increased ryanodine binding affinity of the calcium channel. The Ca2(+)-ATPase activity of the pump was shown to be reduced. The viscosity state of the membranes, assessed by 1,6-diphenyl-1,3,5-hexatriene fluorescence anisotropy, was significantly increased in Mg-deficient membranes. Moreover, these membranes demonstrated an increased content of protein carbonyls as compared with controls. These functional as well as structural changes are closed to those described in sarcoplasmic reticulum membranes oxidatively modified in vitro. Together, these data fitted well with the concept that free radical-induced membrane damages resulting in calcium overload may be at the origin of skeletal muscle lesion during Mg-deficiency. PMID- 8721614 TI - Characterization of drugs as antioxidant prophylactics. AB - There is growing interest in the evaluation of drugs (prescription only medicines and over-the-counter medicines) as antioxidant prophylactics. Although free radical mechanism in human degenerative diseases is now generally recognised, the mechanisms of tissue injury in humans are very complex and it may not be possible to clearly identify the role played by free radicals in the process. This review examines the current evidence to support the notion that drugs for a particular therapeutic category might possess useful antioxidant capacity hence minimising tissue injury due to free radicals. PMID- 8721615 TI - The role of oxidized lipoproteins in atherogenesis. AB - This article reviews our current understanding of the mechanisms of low-density lipoprotein (LDL) oxidation and the potential role of oxidized lipoproteins in atherosclerosis. Studies in hypercholesterolemic animal models indicate that oxidation of LDL is likely to play an important role in atherogenesis. Epidemiological investigations further suggest that the dietary intake of antioxidants is inversely associated with the risk of vascular disease, suggesting that oxidized LDL may be important in human atherosclerosis. By activating inflammatory events, oxidized lipoproteins may contribute to all stages of the atherosclerotic process. Lipoprotein oxidation is promoted by several different systems in vitro, including free and protein-bound metal ions, thiols, reactive oxygen intermediates, lipoxygenase, peroxynitrite, and myeloperoxidase. Intracellular proteins that bind iron or regulate iron metabolism might also play an important role. The physiologically relevant pathways have yet to be identified, however. We assess recent findings on the effects of antioxidants in vivo and suggest potential strategies for inhibiting oxidation in the vessel wall. PMID- 8721616 TI - Horse spleen ferritin inhibits superoxide production by equine blood monocytes in vitro. AB - The effect of horse spleen ferritin (HFR) on the production of superoxide anion (O2.-) by equine blood monocytes was investigated. Preincubation of monocytes with HFR resulted in pronounced inhibition of O2.- production in response to phorbol 12-myristate 13-acetate (PMA), N-formyl-methionyl-leucyl-phenylalanine (FMLP), and opsonized zymosan (OZ). The inhibitory effect of HFR upon stimulation of monocytes with PMA was both dose and time dependent. Maximum inhibition (90%) was observed after preincubation of monocytes with HFR (2 mg/ml) for 18 h before stimulation with PMA. ApoHFR at the same concentration showed only about one third of the inhibitory effect of iron-saturated HFR. Various iron complexes, such as iron dextran, hemin, or ferric ammonium sulfate, had no significant effect on O2.- production by monocytes. Neither catalase (Cat) nor desferrioxamine (DFO) changed the inhibitory effect of HFR. These findings suggest that HFR may play an important role in inhibition of superoxide generation by equine monocytes. Although the mechanism of this inhibition remains unknown, the results obtained suggest that it is not due to ferritin-dependent oxidative inactivation of the NADPH-oxidase system in stimulated monocytes. PMID- 8721618 TI - Reactive oxygen metabolites promote cholesterol crystal formation in model bile: role of lipid peroxidation. AB - In animal models of gallstone disease inflammatory alterations of the gallbladder mucosa are regularly found before the first appearance of cholesterol monohydrate crystals in bile. At sites of inflammation granulocytes generate reactive oxygen metabolites (ROM). The aim of our study was to investigate whether ROM may influence the cholesterol monohydrate crystal formation in supersaturated model bile. Superoxide anions (O2-), hydrogen peroxide (H2O2), and hydroxyl radicals (.OH) were generated by the interaction of Fe(3+)-EDTA with ascorbic acid (Asc). The influence of ROM on cholesterol crystal formation was studied by measurement of the nucleation time. To check whether lipid peroxidation was induced by the ROM generation, production of malondialdehyde equivalents was measured in bile with the thiobarbituric assay. Furthermore, the lipid pattern of bile after ROM exposure was analyzed by thin layer chromatography. Addition of Fe(3+)-EDTA/Asc to model bile markedly decreased the cholesterol nucleation time (NT) (p < 0.01), caused a significant increase in malonidialdehyde equivalents (p < 0.001) and induced the generation of 4-hydroxy-2,3-trans-nonenal (4-HNE). In an attempt to identify a specific oxygen metabolite responsible for the alterations in bile, the effects of various oxygen radical scavengers were tested. Desferal, which prevents -OH generation by chelation of ferrous iron, completely protected bile against Fe(3+)-EDTA/Asc-induced decrease in NT (p < 0.001), increase in lipid peroxidation (p < 0.001) and generation of 4-HNE. Our results indicate that formation of cholesterol crystals in model bile is enhanced by ROM. Hydroxyl radical induced lipid peroxidation appears to be the mechanism responsible for the crystallisation promoting activity of ROM. PMID- 8721617 TI - Expression of stress response genes GADD153, c-jun, and heme oxygenase-1 in H2O2- and O2-resistant fibroblasts. AB - In the present study, the expression of the stress-inducible genes GADD153, c jun, heme oxygenase-1 (HO-1), and HSP70 was compared among parental hamster diploid fibroblasts (HA-1), and cell lines isolated for resistance to either H2O2 (OC14) or O2 (O2R95). Both OC14 and O2R95 cell lines are known to have significantly augmented cellular antioxidant defenses, including increased glutathione content, as well as enhanced catalase, superoxide dismutase, and glutathione peroxidase activities. Northern analysis indicated that basal expression of HO-1 and c-jun is also elevated in these resistant cell lines. Relative to HA-1 values, basal GADD153 mRNA expression was approximately threefold higher in O2R95, but twofold lower in OC14 cells. HSP70 mRNA expression was comparable among parental and resistant cell lines. Both OC14 and O2R95 cells showed greatly enhanced survival following H2O2 exposure. The H2O2 doses that induced 50% toxicity in parental and resistant cells (3 vs. 30-60 x 10(-13) mol/cell, respectively) differed by more than an order of magnitude. Similarly, GADD153, c-jun, and HO-1 mRNA were elevated in control cells following exposure to doses of H2O2 an order of magnitude lower than is required for gene activation in resistant cell lines. Nonetheless, at equitoxic doses, the level of induction of GADD153 and HO-1 was greater in resistant than in parental cell lines. Taken together, our results suggest that alterations in the basal level of expression of certain stress-responsive genes, including HO-1, c-jun, and GADD153, may contribute to the H2O2-resistant phenotype in these fibroblast cell lines. Further, changes in the regulation of these genes in response to adverse stimuli may provide an additional mechanism for enhanced cell survival following oxidative stress. PMID- 8721619 TI - Effects of hydroxyl radical scavengers on relaxation of supercoiled DNA by aminomethyl-trimethyl-psoralen and monochromatic UVA photons. AB - The ability of scavengers of hydroxyl radical (OH radical) to modulate the photosensitized relaxation (induction of the first single-strand break) of supercoiled plasmid DNA with UVA photoactivated 4'-aminomethyl-4,5',8 trimethylpsoralen was examined by comparing the dose reduction factor (DRF: the ratio of fluence required to induce the same degree of relaxation in the absence to the presence of OH radical scavengers). The addition of mannitol, azide, acetate, or formate at concentrations inversely proportional to the value of the rate constants for the scavenging of OH radicals partially attenuated the supercoiled DNA relaxation. The degrees of protection afforded by the four scavengers in the presence of AMT photoactivated by either 334 nm or 365 nm monochromatic photons were similar, giving an average DRF of about 0.25 in all cases. Given the diverse chemical nature of the scavengers and their wide range of concentrations utilized, these findings are evidence for the involvement of a Type I photosensitization in the induction of DNA single-strand breaks by photoactivated AMT. PMID- 8721620 TI - Increased plasma lipid peroxidation in patients with aceruloplasminemia. AB - Aceruloplasminemia is a newly recognized autosomal recessive disorder of iron metabolism due to mutations in the ceruloplasmin gene. Although the presence of these mutations reveals an essential role for ceruloplasmin in human biology, the mechanisms of tissue injury in this disease are unknown. We report here on the identification of increased plasma lipid peroxidation in multiple affected family members with aceruloplasminemia. Consistent with the absence of serum ceruloplasmin, plasma ferroxidase activity was markedly reduced and serum ferritin was significantly increased. Plasma lipid peroxidation was determined as thiobarbituric acid-reactive products (TBA products) in plasma samples from control, heterozygote, and affected patients. Basal levels of lipid peroxides were three times control values in patients with aceruloplasminemia and were significantly increased in these patients in the presence of copper ions and hydrogen peroxide. In each case these increases were suppressed by the addition of exogenous ceruloplasmin. These data suggest that increased susceptibility to lipid peroxidation may contribute to the unique neuropathology observed in patients with aceruloplasminemia and imply a role for free radical-mediated tissue injury in degenerative disorders of the basal ganglia. PMID- 8721621 TI - The isolation, purification and identification of fumitremorgin B produced by Aspergillus fumigatus. AB - Twenty-six strains of Aspergillus fumigatus were screened for toxigenicity for fumitremorgins A and B. Twenty-three of 26 strains can produce fumitremorgin B in rice medium determined by TLC and HPLC, and no fumitremorgin A was detected. The strains of no. C4104 and no. 3656 were inoculated onto 5 kg of rice media and incubated in a modified procedure. Finally, 4.0 g of fumitremorgin B was obtained after extraction and purification by modified methods, and was confirmed by TLC, HPLC, spectral analysis together with other physicochemical analysis. This is the first report of the preparation of fumitremorgin B in China. PMID- 8721622 TI - The preparation and identification of fumitremorgin B-hemisuccinate-carrier proteins. AB - A method for the preparation and identification of fumitremorgin B-hemisuccinate carrier protein is described. The overall yield of the fumitremorgin B hemisuccinate (FTBS) after final purification was 78.6%. The FTBS was characterized by UV, IR, EIMS, element analysis, and 1H, 13C NMR. IR was also used to determine the formation of complete antigen complexes. The reaction route was analysized. PMID- 8721623 TI - Production and characterization of a monoclonal antibody against sterigmatocystin. AB - A monoclonal antibody, McAb-ST, specific for sterigmatocystin (ST) was produced in vitro and in vivo by a hybridoma anti-ST1. Anti-ST1 was prepared by fusion of SP2/0 murine myeloma cells with spleen cells isolated from BALB/c mouse that had been immunized with a complete antigen BSA-ST (ST conjugated to bovine serum albumin). Analysis revealed that McAb-ST was of the IgG1 type, the molecular weight of McAb-ST was 168,000 daltons (53,000 for each heavy chain, 31,000 for each light chain), and the apparent affinity constant of McAb-ST with H-ST (Hemocyanin and ST conjugate) was 1.29 x 10(9) mol/L. The relative cross reactivities of McAb-ST with ST analogs were less than 2% when determined by a competitive indirect enzyme-linked immunosorbent assay (CI-ELISA). The minimal 0.5 pg of ST in 50 microliters of solution could be detected with CI-ELISA using McAb-ST, with a linear range of 0.1-10 ng/mL. PMID- 8721624 TI - Involvement of mercury in platelet aggregation--probable mechanism of action. AB - Both in vivo and in vitro Hg2+ caused dose dependent inhibition of rat platelet aggregation. However, as low a dose as 88.8 pM Hg2+ could initiate ADP-induced aggregation even in the absence of added calcium. The action of Hg2+ was found to be unaffected by antagonists such as cAMP and theophylline either alone or in combination. Hg2+ induced platelet aggregation was inhibited completely by pre incubation with aspirin (1 mmol.L-1), but it was ineffective when added after Hg2+. The results also indicate that the normal platelet function is affected by both high and low concentrations of Hg2+. PMID- 8721625 TI - Human sperm chromosome analysis--study on human sperm chromosome mutagenesis induced by carbon disulfide. AB - The aim of this study was to investigate the effect CS2 of on human sperm chromosomal aberration. The human sperm/hamster egg fusion technique was used to analyze 203 human sperm chromosome complement from 9 healthy volunteers. The incidence of numerical aberration was 1.0% and that of structural chromosome aberration was 5.9% and total abnormalities was 6.9%. Structural aberrations consisted of breaks, deletions, centric rings, fragments, and chromatid exchange. The results from high concentration group (10 mumol.L-1 CS2) showed that the incidence of chromosomal aberration rate was significantly higher than that of the control group. The results indicate that high concentration of CS2 might directly cause mutagenesis of the germ cell. PMID- 8721626 TI - Blood lead dynamics of lead-exposed pregnant women and its effects on fetus development. AB - The dynamics of blood lead (Pb-B) and blood zinc protoporphyrin (ZPP-B) of women in early pregnancy and parturient women with lead exposure and the effects on fetus development were investigated. Pb-B of lead-exposed women was high: 0.984 mumol/L (20.38 micrograms/dl) and ZPP was 84.52 micrograms/dl. Cord blood Pb-B was 0.896 mumol/L (18.56 micrograms/dl) and cord blood ZPP was 69.24 micrograms/dl. In the control group, Pb-B was 0.261 mumol/L (5.41 micrograms/dl), ZPP-B, 37.59 micrograms/dl, cord blood, Pb-B 0.34 mumol/L (7.03 micrograms/dl), and cord ZPP-B 49.0 micrograms/dl. There was a significant correlation between blood lead and blood ZPP, maternal Pb-B and cord Pb-B, maternal Pb-B and cord ZPP B. The significance of the consistency of high level Pb-B and the effects on fetus development is discussed. PMID- 8721627 TI - Promoting activity of microcystins extracted from waterblooms in SHE cell transformation assay. AB - Microcystis aeruginosa is the dominant algae in most of the eutrophicated lakes in China. It can produce cyclic heptapeptides, known as microcystins, which can cause liver damage in wild and domestic animals. In this paper, a two-stage transformation assay for demonstrating the carcinogenic effects of the algal toxins is reported. The cell strain used in this assay was derived from embryos of Syrian golden hamster and the algal toxins were extracted from Microcystis aeruginosa, termed microcystis raw toxin (MRT). To elucidate its promoting activity, the target cells were first exposed to a low dosage of 3 methylcholanthrene (MCA) and then to MRT. The results showed that MRT significantly enhanced the MCA initiated cell transformation, and a dose-response relationship was observed, but it failed to induce transformation of SHE cells not pretreated by MCA. These results suggest that the MRT play an important role in the malignant transformation of SHE cells. MRT may thus be a tumor promoter, and this transformation assay with SHE cells may be used to predict tumor prompting activity of environmental chemicals, before long-term in vivo two-stage carcinogenesis experiments are carried out. PMID- 8721628 TI - Role of phosphatases during transport and energy metabolism in Labeo rohita after exposure to cypermethrin. AB - Freshwater fish, Labeo rohita, were exposed to sublethal concentration (0.5 micrograms.L-1) of cypermethrin for 7 and 15 days to examine the bioenergetics in functionally four different tissues, namely, gill, liver, brain and muscle. Whole animal oxygen consumption was measured first and it was found to decrease in both the exposure periods (EPs), manifesting respiratory distress of the animal in toxic environment. Ionic regulation and energy requirements were also found to be altered under stress, as observed by the inhibition of both Na+/K+ and Mg2+ ATPases at 7d EP and elevation at 15d EP. Increase in glucose-6-phosphate dehydrogenase (G-6-PDH) was consistent with the increase in exposure time. Attenuation of acid and alkaline phosphatases were noticed in treated fish after 7 days but were close to normalcy at 15d EP. These results clearly indicate that the fish were affected at 7d EP but adapted to the toxic environment within 15 days. It shows that at this concentration cypermethrin is only moderately toxic and the animal has alternate pathways to derive energy and survive. PMID- 8721629 TI - Histological studies of atrazine toxicity on the thyroid gland in rats. AB - Little is known about the toxic activity of the atrazine (a herbicide, commonly used in agricultural production) on the thyroid gland. In this study the compound was administered orally in female albino rats at sublethal exposure equivalent to 0.2 LD50 doses for 6 and 12 days. At termination of dosing the anesthetized animals were killed and blood was drawn for the determination of serum triiodothyronine (T3) and thyroxin (T4). A dose-dependent decrease of serum T3 concentration was observed in all the groups (control: 0.57 nmol-L-1; 6 days: 0.35 nmol-L-1; 12 days: 0.21 nmol-L-1). The thyroid gland was examined light microscopically. Bouin's solution-fixed thyroids were embedded in paraffin and sections cut at 6 microns, stained separately with toluidine blue according to Slinchenko's method. Histologically in experimental groups epithelium featured small cuboidal cells and occasional structures of the follicles confluence within epitheliomers. A dose-dependent changes of the following parameters were observed: (a) increasing of number of follicle-building thyroid cells; (b) increasing of follicular volume; (c) decreasing of nucleus volume. Investigation of the whole population of thyroid mast cells disclosed no change in degranulation intensity. By contrast, degranulation intensity was decreased in perifollicular mast cells from groups treated with atrazine in dose-dependent manner. There are no changes observed in degranulation of stromal mast cells. These results suggesting that differences in response to the atrazine might account for an aspect of the functional heterogeneity within the rat thyroid mast cell population. PMID- 8721630 TI - The inhibitory effect of extracts from Fructus lycii and Rhizoma polygonati on in vitro DNA breakage by alternariol. AB - Alternariol caused DNA single-strand breakage. Conversion of the closed circular double-stranded supercoiled DNA (pBR 322) to the nicked circular form and linear form was used to investigate the effect of extracts of some Chinese medical herbs on DNA nicking induced by alternariol. Some substances in the extracts of Rhizoma polygonati (RP) and Fructus lycii (FL) were shown to protect DNA from the attack by alternariol. Some substance in the RP may bind to plasmid DNA, and this binding reduces the electrophoretic mobility of DNA. These results indicate that substances from FL and RP may be used as DNA protectors. It is possible that they play an important role in preventing cancer. PMID- 8721631 TI - Protective effect of vitamin E on liver damage induced by 2-chloro-1, 3 butadiene. AB - The present study was performed to determine the influence of lipid peroxidation and perturbance of Ca2+ homeostasis on liver damage induced by 2-chloro-1, 3 butadiene (CBD) and the protective effects of vitamin E in Wistar rats. Animals were given intraperitoneally different doses (8,40 or 200 mg.kg.-1 daily) of CBD for 21 days, and the following dose-dependent events were observed: liver damage, significant increase in liver lipid peroxides, and decreases in activities of erythrocytic glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD). The pretreatment of rats with vitamin E (po 150 mg.kg-1) before administering CBD (ip 60 mg.kg-1) daily for 21 days prevented the following CBD-induced changes, the increase in serum cholyglycine (CG), hepatic LP, hepatic mitochondrion LP, hepatic oxidized glutathione (GSSG) (while the significant increase of reduced glutathione (GSH) was not affected) and the decrease in activities of erythrocytic SOD and hepatic mitochondrial calcium sequestration. These results suggest that lipid peroxidation and perturbance of Ca2+ homeostasis appear to contribute to the hepatotoxicity of CBD, and vitamin E might prevent the liver damage induced by CBD. The decrease in activities of GSH-Px and SOD in erythrocytes might be used as biomarkers for adverse effects of CBD on defense system against lipid peroxidation. PMID- 8721632 TI - Influence of percutaneous transluminal coronary rotational atherectomy with adjunctive percutaneous transluminal coronary angioplasty on coronary blood flow. AB - The purpose of this study was to examine the influence of sequential percutaneous transluminal coronary rotational atherectomy (PTCRA) and coronary angioplasty on coronary blood flow reserve in patients. Rotational coronary atherectomy restores lumen patency by partially ablating fibrocalcific plaque, releasing microparticulate debris into the distal coronary circulation. Adjunctive balloon angioplasty is usually performed to optimize the angiographic luminal dimensions. Serial alterations in coronary physiology have not been reported. Fourteen lesions in 13 patients were treated by sequential rotational atherectomy followed by adjunctive balloon angioplasty. Poststenotic baseline coronary blood flow velocity was measured by using a Doppler flow wire (FloWire, Cardiometrics, Inc., Mountain View, Calif.), and coronary blood flow was calculated by using the distal vessel cross-sectional area obtained by quantitative coronary angiography. Data were acquired at baseline and during hyperemia (12 to 18 microg of intracoronary adenosine), before and after PTCRA, and again after balloon angioplasty. The mean stenosis decreased from 76 percent +/- 12 percent at baseline to 21 percent +/- 11 percent at the completion of the procedure (p<0.01). The minimal luminal diameter (by quantitative coronary angiography) was 0.7 +/- 0.4 mm at baseline, increased to 1.9 +/- 0.4 mm after rotational atherectomy (p<0.01), and increased to 2.4 +/- 0.5 mm after balloon angioplasty (p<0.01 versus baseline and PTCRA). Distal (poststenotic) coronary blood flow at baseline was 47 +/- 23 ml/min and 57 +/- 38 ml/min during hyperemia. After PTCRA, coronary blood flow increased to 104 +/- 59 ml/min and to 132 +/- 73 ml/min with hyperemia. After adjunctive angioplasty, coronary blood flow was 84 +/- 40 ml/min (p=not significant [NS] vs PTCRA) and increased to 143 +/- 81 ml/min with hyperemia (p=NS vs PTCRA). The poststenotic coronary flow reserve increased from an initial value of 1.1 +/- 0.2 ml/min to 1.3 +/- 0.3 ml/min after PTCRA (p=NS vs baseline) and to 1.6 +/- 0.3 ml/min after adjunctive balloon angioplasty (p<0.01 vs p=NS vs PTCRA). PTCRA significantly increased resting coronary blood flow. Adjunctive balloon angioplasty did not significantly augment resting or hyperemic coronary blood flow more than that achieved by rotational atherectomy alone. These data demonstrate that PTCRA alone improves baseline coronary blood flow with minimal additional physiologic change after adjunctive balloon angioplasty. PMID- 8721633 TI - High-speed rotational atherectomy: six-month serial quantitative coronary angiographic follow-up. AB - One hundred twenty-three patients treated with high-speed rotational atherectomy (HSRA) were restudied 6.9 +/- 1.2 months later. At the follow-up, the number of focal concentric lesions increased from 32.2 percent to 63.0 percent, p<0.01, with decrease of type C lesions from 54.8 percent to 30.8 percent, p<0.05. Comparison of the degree of the net gain (NG) showed more severe baseline lesions in the high-gain group (NG >20 percent) compared with the moderate-gain group (20 percent > NG > 0 percent) and to the loss group (minimal luminal diameter [MLD] 0.8 +/- 0.4 mm vs 1.0 +/ 0.4 mm, p<0.05; and 1.2 +/- 0.5 mm; p<0.01, respectively). Highest initial gain (36.5 percent +/- 26.2 percent vs 24.5 percent +/- 18.1 percent; p<0.015; and 19.0 percent +/- 23.2 percent; p<0.001) as well as lowest late loss (1.8 percent +/- 21.7 percent vs 14.0 percent +/-18.4 percent; p<0.01 and 28.1 percent +/- 25.0 percent; p<0.01) were found in the high NG group. A higher interaction between burr and atheroma resulted in the lowest restenosis rate of 6 percent. PMID- 8721634 TI - Absence of Q waves after thrombolysis predicts more rapid improvement of regional left ventricular dysfunction. AB - Although the natural history of regional left ventricular (LV) dysfunction after Q-wave and non-Q-wave myocardial infarction (MI) was well defined in the prethrombolytic era, the functional and structural implications of the absence of Q waves after thrombolysis are less clear. Echocardiography was performed within 48 hours of admission (entry) in 86 patients treated with thrombolysis for their first MI. The extent of abnormal wall motion (AWM; square centimeters) and LV endocardial surface area index (ESA; square centimeters per square meters) were quantified by using a previously validated echocardiographic endocardial surface mapping technique. Electrocardiography (ECG) performed at 48 hours after thrombolysis was used to classify patients into groups with (Q; n=70) and without (non-Q; n=16) Q waves. All patients in the Q group had regional LV dysfunction on initial echocardiogram compared with 69 percent of those in the non-Q group (p<0.001). When the patients in the non-Q group without AWM were excluded from analysis, there was no significant difference in the extent of AWM between the Q and non-Q groups. Among those patients with AWM on entry, follow-up echocardiography at 6 to 12 weeks demonstrated a significant reduction in extent of AWM for both the Q and non-Q groups. However, the fractional change in AWM was significantly greater in the non-Q than in the Q group (-0.74 +/- 0.28 vs -0.29 +/- 0.44; p<0.02), with a trend toward less AWM at follow-up in the non-Q than in the Q group. The mean ESAi was not significantly different between the two groups at entry or at follow-up. In conclusion, failure to develop Q waves after thrombolysis predicts a lower likelihood of developing regional LV dysfunction and, when such dysfunction is present, predicts a greater degree of recovery. PMID- 8721635 TI - Comparison of thickness and distensibility in the carotid artery and descending thoracic aorta: in vivo ultrasound assessment. AB - Early atherosclerotic changes in the carotid artery and thoracic aorta have been examined by high-frequency ultrasound measuring of intimal-medial thickness and stiffness. Whether changes in stiffness and thickness occur in parallel and whether the determinants of stiffness and thickness in the two vessels are similar is unknown. To examine the relation between ultrasonographic measures of atherosclerosis in the carotid and the thoracic aorta, 146 patients aged 20 to 84 years (mean 54 years) were studied by both transesophageal echocardiography (TEE) and carotid duplex scanning. From two-dimensionally derived M-mode recordings of the thoracic aorta and high-frequency B-mode imaging of the common carotid, the intimal-medial thickness was measured along with diastolic and systolic diameters for calculation of stiffness. Interobserver and intraobserver variability of carotid and aortic intimal-medial thickness and diameter were low. There was a good relation between carotid and aortic intimal-medial thickness (r = 0.69; p=0.0001). Age was the major independent determinant of thickness in both vessels. Carotid and aortic stiffness, as measured by Peterson's elastic modulus, were less closely related (r=0.51; p=0.0001). Age was the only independent predictor of stiffness in both vessels. In conclusion, structural ultrasonographic manifestations of early atherosclerosis in the carotid artery and thoracic aorta are closely related. Large population studies measuring only carotid intimal-medial two-thickness may reflect atherosclerotic changes occurring throughout the vascular bed. PMID- 8721636 TI - Autoantibody titers to oxidized low-density lipoprotein in patients with coronary atherosclerosis. AB - Oxidation of low-density lipoprotein (LDL) is considered to be the initial step in the atherosclerotic process. Autoantibodies to oxidized LDL (ox-LDL) have been detected in human serum. We used an enzyme-linked immunosorbent assay technique to measure autoantibody titers in 63 normal subjects and patients with coronary artery disease. Thirty-five patients underwent coronary angiography for suspected coronary artery disease. Patients were divided into the following categories: group 1, 20 healthy young volunteers; group 2, 8 patients age-matched to the catheterization patients; group 3, 10 patients with normal coronary angiograms; and group 4, 25 patients with angiographic coronary artery disease. Autoantibody titers to ox-LDL were group 1, 0.142 +/- 0.023; group 2, 0.197 +/- 0.039; group 3, 0.183 +/- 0.038; and group 4, 0.340 +/- 0.026. There was no statistical difference among groups 1, 2, and 3, but the difference between these groups and group 4 was highly significant (p < 0.05). This study demonstrates that (1) autoantibodies to ox-LDL can be detected in normal subjects and in patients with abnormal coronary angiograms and (2) significantly higher titers of autoantibodies to ox-LDL were seen in patients with angiographic evidence of coronary artery disease. PMID- 8721637 TI - Assessment of reversible dyssynergic segments after acute myocardial infarction: Dobutamine echocardiography versus thallium-201 single photon emission computed tomography. AB - Only a moderate degree of concordance has been reported between stress redistribution-reinjection thallium-201 single photon emission computed tomography (SPECT) and dobutamine echocardiography for the identification of myocardial viability after acute myocardial infarction. SPECT with rest reinjection performed 4 hours after exercise testing and digitized two dimensional (2-D) ultrasound reconstruction of the left ventricle at baseline and after low-dose dobutamine (5 to 10 microg/kg/min) infusion were compared in 50 patients > or = 8 days (12 +/- 7 days) after acute myocardial infarction. Five patients were excluded because of technically inadequate echocardiograms. Both SPECT and dobutamine echocardiography were assessed in a 16-segment model and interpreted in the remaining 45 patients. Digitized 2-D reconstruction of the left ventricle in each wall motion was scored from 1 (normal) to 4 (dyskinesia). Myocardial viability was identified on ultrasound wall-motion improvement of one or more grades from baseline to echocardiography performed > or = 30 days (60 +/- 41 days) after systematic revascularization procedure of the infarct-related artery. Reversible defect under thallium-201 SPECT and wall-motion improvement under dobutamine echocardiography were concordant in 163 (69 percent) of the 235 baseline dyssynergic segments and in 30 (67 percent) patients. Myocardial viability was identified after angioplasty (n=37) or surgery (n=8) in 29 patients and 109 segments. Positive and negative predictive values per patient in the diagnosis of myocardial viability were 86 percent and 57 percent, respectively, for stress thallium-201 SPECT with reinjection, and 100 percent and 62 percent for dobutamine echocardiography. Positive and negative predictive values per segment were 80 percent and 69 percent for the isotopic method and 91 percent and 70 percent for dobutamine echocardiography. We conclude that dobutamine echocardiography and stress thallium-201 SPECT with reinjection have similar accuracies to identify myocardial viability after acute myocardial infarction, with excellent positive but relatively low negative predictive values. PMID- 8721638 TI - Increased plasma adrenomedullin in acute myocardial infarction. AB - Adrenomedullin has a potent vasodilating effect comparable to that of calcitonin gene-related peptide. To investigate the pathophysiologic role of endogenous adrenomedullin, we determined sequentially the plasma adrenomedullin level in 15 consecutive patients with acute myocardial infarction (AMI). Plasma adrenomedullin was higher immediately after the onset of AMI and decreased gradually; plasma levels during the 3-week period after the AMI were higher than plasma levels in 15 healthy control subjects (p < 0.001), with higher levels in patients with congestive heart failure than in patients without congestive heart failure throughout the period of the study (p < 0.05). Plasma adrenomedullin was positively correlated with pulmonary capillary wedge pressure, pulmonary arterial pressure, right atrial pressure, and heart rate in the early stage of AMI. These findings suggest that the elevation of plasma adrenomedullin is related to the retention of body fluid volume, the enhancement of sympathetic activity, and/or the elevation of pressure in pulmonary vascular beds. Adrenomedullin may act against excessive vasoconstrictors increased in AMI. PMID- 8721639 TI - Limiting lipid peroxidation in the nonischemic zone of infarcted rat hearts by indomethacin improves left ventricular function without affecting myocardial healing and remodeling. AB - We recently showed that indomethacin reduces lipoperoxidation occurring in the nonischemic zone (right ventricle + septum; RVS) of rat hearts with permanent coronary occlusion. The purpose of this study was to determine whether this biochemical effect is associated with an improvement of residual cardiac function or with modifications of delayed ventricular remodeling, or both. Rats received either indomethacin (1 mg/kg i.p.) or a placebo 5 min before coronary occlusion, and then twice a day for 48 hours. Six hours after ligation, myeloperoxidase (MPO) activity (leukocyte infiltration index) was measured in the ischemic zone (left ventricle; LV) and in RVS. Forty-eight hours after ligation, left ventricular function was assessed in vivo. Alterations in myocardial geometry were estimated 21 days after ligation on histologic cross-sections. In the indomethacin group, cardiac function was improved compared with that with placebo, and the indexes of ventricular remodeling were not affected by the treatment. Finally, indomethacin at the dose used did not affect MPO activity in the LV. It is concluded that indomethacin treatment leads to a better maintenance of residual nonischemic tissue contractility without worsening late ventricular shape changes. These results could in part be explained by the ability of the treatment to decrease inflammatory mediator-induced oxidative stress in RVS without affecting infarct healing caused by leukocyte infiltration in LV. PMID- 8721640 TI - Adaptation to myocardial ischemia during repeated dynamic exercise in relation to findings at cardiac catheterization. AB - It has been suggested that the myocardium is able to recruit endogenous protective mechanisms in response to repeated ischemia and reperfusion. We set out to study whether this is manifested in patients with coronary artery disease in the form of fewer signs of myocardial ischemia during the second of two successive exercise tests and whether any relations exist between ischemia adaptation and findings at cardiac catheterization. Twenty-one patients with typical angina pectoris symptoms underwent two repeated bicycle exercise tests with identical protocols, followed by cardiac catheterization and coronary angiography the next day. The first exercise test was discontinued whenever a 2 mm ST depression in the electrocardiogram (ECG) was achieved or further exercise was limited by symptoms. The second exercise test was performed after disappearance of the symptoms or ST depression or both. Kaplan-Meier survival analysis for the appearance of a 1 mm ST depression demonstrated improved ischemia tolerance during the second test, when the required time for its appearance was significantly longer (6.5 +/- 0.8 min vs 4.5 +/- 0.5 min; p = 0.005). The maximal intensity of anginal pain was lower during the second exercise (2.2 +/- 1.0 min vs 0.7 +/- 0.3 min in Borg's scale; p < 0.001), and the time required for disappearance of the ST depression was shorter after this exercise (3.0 +/- 0.8 min vs 6.2 +/- 0.9 min; p = 0.003), with a similar tendency in the disappearance of angina. The rate-pressure product on the appearance of a 1 mm ST depression was significantly higher during the second test (17,990 +/- 1210 mm Hg x min-1 vs 15,960 +/- 869 mm Hg x min-1; p = 0.009). Eighteen of the patients had three-vessel disease, as evidenced by coronary angiography, and the change in the time required for the appearance of a 1 mm ST depression in the repeated exercise tests was inversely correlated with the severity of the left anterior descending (LAD) coronary artery obstruction (r = -0.61; p = 0.006) and left ventricular end-diastolic pressure (r = -0.50; p = 0.03). No significant correlation with the degree of collateral vessels was found. We conclude that most patients with extensive coronary artery disease are able to increase their tolerance of ischemia during repeated dynamic exercise and that increased vasodilation and oxygen delivery are the major mechanisms for this warm-up phenomenon. On the other hand, collaterals visible in routine resting anglography do not predict the degree of adaptation to ischemia during repeated dynamic exercise. PMID- 8721641 TI - Effect of beta-blockade on dobutamine stress echocardiography. AB - Dobutamine is an effective pharmacologic stress used in conjunction with echocardiography because of its beta-agonist properties. Concurrent beta-blockade might alter this effectiveness; however, current clinical experience has been variable. The purpose of this study is to determine whether concurrent beta blockade alters the ability of a dobutamine stress echocardiogram to detect a fixed coronary stenosis by preventing the onset of a wall motion abnormality or by altering the dose at which the wall motion abnormality appears. Paired dobutamine stress tests with and without beta-blockade (esmolol 500 microgram/kg initial bolus, 100 microg/kg/min infusion) were performed in a canine model (n = 8) with a fixed single-vessel coronary stenosis. Heart rate, systolic pressure, proximal left anterior descending coronary flow, myocardial thickening (by sonomicrometry), and left ventricular area change (by epicardial echocardiography) were monitored. Simultaneous beta-blockade resulted in (1) a delayed and diminished increase in hemodynamic parameters (peak heart rate 164.1 +/- 22.3 without beta-blockade vs 110.1 +/- 28.9 beats/min with beta-blockade, p < 0.001, and peak systolic blood pressure 137.9 +/- 26.8 mm Hg without beta blockade vs 107.3 +/- 15.3 mm Hg with beta-blockade, p = 0.01), (2) an elimination of the physiologic effects of low-dose (5 and 10 microg/kg/min) dobutamine (-0.7 percent +/- 16.7 percent change in myocardial thickening from baseline with beta-blockade, p = NS), and (3) an elimination or alteration in timing of inducible wall motion abnormalities caused by severe coronary artery stenoses (mean termination dose 28.8 +/- 9.9 with beta-blockade vs 15.6 +/- 6.1 microg/kg/min without beta-blocker, p < 0.01). The findings in this canine model suggest that the competitive antagonist markedly attenuates the ability of dobutamine stress echocardiography to detect a significant coronary lesion and may alter its ability to detect viable myocardium at low-dose testing. Further clinical studies to determine the sensitivity of dobutamine stress echocardiography in the presence of beta-blockers and to establish protocol standards are necessary. PMID- 8721642 TI - Gender-related differences in exercise ventricular function among healthy subjects and patients. AB - Increasing numbers of women are undergoing noninvasive stress testing for coronary artery disease evaluation. Limited information is available regarding the presence, magnitude, and importance of gender-related differences in exercise ventriculography among the heterogeneous population of patients referred for noninvasive stress testing. Patients referred for exercise radionuclide ventriculography between 1979 and 1986 were evaluated, including 175 patients with a likelihood of coronary artery disease, 59 patients with angiographically normal coronary arteries, and 419 patients with coronary artery disease. Overall, women demonstrated higher resting left ventricular ejection fraction and lower delta left ventricular ejection fraction response to exercise compared with men. Although left ventricular response to exercise correlated with the underlying severity of coronary artery disease in both women and men, fewer women demonstrated a delta left ventricular ejection fraction >5 percent despite a lower prevalence of multivessel coronary artery disease compared with men. We conclude that gender-related differences in left ventricular response to exercise are present in a wide range of patients referred for testing. PMID- 8721643 TI - Geriatric patients with acute myocardial infarction: Cardiac risk factor profiles, presentation, thrombolysis, coronary interventions, and prognosis. AB - Elderly patients have a higher mortality after acute myocardial infarction (MI) yet are treated less aggressively than younger patients. To determine (l) the risk-factor profiles, (2) presentation, (3) management, and (4) hospital outcomes for the elderly (> or = 75 years) compared with middle aged (66 to 74 years) and younger (< or = 65 years) patients in the 1990s, we studied 561 consecutive patients with acute MI. Compared with younger patients, the elderly more frequently had congestive heart failure (40 percent vs 14 percent; p < 0.00001) and non-Q wave infarctions (76 percent vs 56 percent; p < 0.005), received thrombolysis (9 percent vs 34 percent; p < 0.0001), and underwent catheterization (35 percent vs 73 percent; p < 0.00001), percutaneous transluminal coronary angioplasty (9 percent vs 31 percent; p < 0.0002), and coronary artery bypass grafting (5 percent vs 15 percent; p < 0.03) less frequently. Those who did not receive thrombolysis all had contraindications. Mortality was higher in the elderly (19 percent vs 5 percent; p < 0.004), especially among those who did not receive thrombolysis (20 percent vs 7 percent; p < 0.03). Multivariate predictors of mortality included age, and congestive heart failure. In addition, when clinical course and management variables were considered, use of the intraaortic balloon pump was a predictor of mortality, whereas undergoing coronary angiography was a negative predictor (relative risk, 0.3; 95 percent confidence intervals, 0.1 to 0.6). PMID- 8721644 TI - Atrioventricular node properties in patients with accessory pathways. AB - A study during the era of surgical ablation suggested that atrioventricular (AV) nodal conduction is faster in patients with accessory pathways than in controls. In the present study, AV nodal characteristics were studied in 30 patients who underwent radiofrequency ablation of an accessory pathway and compared to 23 control patients. Sinus cycle length, AH and HV intervals, AV block cycle length, ventriculoatrial (VA) block cycle length, AV nodal effective refractory period, and VA effective refractory periods were measured in control and postablation accessory pathway patients before and after autonomic blockade with 0.04 mg/kg of atropine and 0.2 mg/kg of propranolol. The mean sinus cycle length in the control and accessory pathway groups did not differ significantly at baseline (798 +/- 211 and 766 +/- 156 msec, respectively) or after autonomic blockade (654 +/- 98 and 649 +/- 108 msec, respectively). The mean AH interval in the accessory pathway group (77 +/- 15 msec) was significantly shorter than in the control group (91 +/- 22 msec; p < 0.05) at baseline; however, there was no difference after autonomic blockade. No other significant differences were observed between the accessory pathway and control groups. These results demonstrate that AV nodal properties of patients with accessory pathways are not significantly different from controls and suggest that previously reported differences may have been due to selection bias. PMID- 8721645 TI - Characteristics of bipolar electrograms during anterograde mapping: the importance of accessory atrioventricular pathway location. AB - Local endocardial electrograms recorded at successful radiofrequency ablation sites during anterograde mapping (QRS-V interval, atrioventricular [AV] interval, AV ratio, the presence of accessory pathway potential, and stability of atrial and ventricular electrogram) were analyzed and compared according to location of accessory pathways. The mitral and tricuspid annuli were divided into 10 regions. Endocardial electrograms differed with regard to location of accessory AV pathways. The QRS-V interval was more negative in right posteroseptal, right free wall and right anteroseptal locations than the interval in other locations. The AV interval was longer in posteroseptal than the interval in left and right free wall sites. The stability of atrial and ventricular electrograms was better in left free wall sites than in posteroseptal sites and worst in right free wall and right anteroseptal sites. The variation in local electrograms at successful ablation sites with respect to pathway location may partly explain the low predictive value and the marked variation in previously suggested criteria for selecting target sites for radiofrequency energy delivery. The results also imply that the influence of accessory pathway location must be considered when attempts are made to establish electrogram-based criteria for predicting successful ablation of accessory pathways. PMID- 8721646 TI - Fibrinogen and fibrin D-dimer levels in paroxysmal atrial fibrillation: evidence for intermediate elevated levels of intravascular thrombogenesis. AB - Because abnormalities in hemostatic factors may in part account for the risk of stroke and thromboembolism in atrial fibrillation, we measured plasma fibrinogen and fibrin D-dimer levels in 33 patients (18 men and 15 women, mean age 60.8 +/- 1.4 years [mean +/- SEM]) with paroxysmal atrial fibrillation (PAF) and 12 patients (3 men and 9 women, mean age 51.0 +/- 4.2 years) with paroxysmal supraventricular tachycardia (PSVT). Levels of these markers were compared to levels in (1) patients with chronic atrial fibrillation; (2) hospital controls (age-matched [age +/- 5 years] and sex-matched patients in sinus rhythm with coronary artery disease and normal left ventricular function); and (3) healthy population controls in sinus rhythm. Patients with PAF had intermediate levels of median plasma fibrinogen and fibrin D-dimer when compared to patients with chronic atrial fibrillation and controls in sinus rhythm (both p < 0.001). There was no relation with atrial size or ventricular function on echocardiography. Patients with PSVT had plasma fibrinogen and fibrin D-dimer levels that were similar to the median levels of the population controls, suggesting that there was no excess in thrombogenesis. These findings are consistent with the hypothesis that atrial fibrillation is related to the increases in plasma fibrinogen and fibrin D-dimer levels. Patients with PAF have intermediate levels of these markers, a finding that is consistent with the intermediate risk of thromboembolism in such patients. PMID- 8721647 TI - Prevalence, resolution, and determinants of late potentials in patients with unstable angina and left ventricular wall motion abnormalities. AB - Although transient myocardial ischemia such as exercise-induced ischemia has not been reported to be associated with the occurrence of late potentials, the association of late potentials with more profound ischemic damage, which is represented by reversible but prolonged left ventricular wall motion abnormalities, has not been demonstrated. We prospectively evaluated 37 unstable angina patients who had reversible but prolonged wall motion abnormalities after resolution of chest pain and electrocardiogram (ECG) changes. Signal-averaged ECG (SAECG) and echocardiogram were recorded during the acute phase and before hospital discharge. Late potentials were present in 6 (16 percent) patients on the initial SAECG recording and resolved in all 6 patients on the second recording before hospital discharge. Normalization of inferior left ventricular wall motion abnormality and multivessel disease were observed more frequently in patients with late potentials on the initial recording than in patients without (p < 0.05 and p < 0.05, respectively). In conclusion, late potentials were observed in patients who had reversible but prolonged wall motion abnormalities; these late potentials were resolved with improvement of left ventricular wall motion abnormalities. These results suggest that myocardial ischemia with prolonged wall motion abnormalities is a possible mechanism of the occurrence of late potentials. PMID- 8721648 TI - Clinical use of posterior electrocardiographic leads: a prospective electrocardiographic analysis during coronary occlusion. AB - Posterior electrocardiographic leads have been described for quite a while; however, their use in diagnosing acute posterior wall myocardial infarction and identifying infarct-related arteries has not been well used. We prospectively studied electrocardiographic changes during balloon occlusion of single-vessel right coronary artery (RCA) and circumflex coronary arteries (LCX). Thirty four inflations were performed in RCAs and 38 in LCXS. Analysis of the patients with ECG changes revealed that the most common ECG change during RCA occlusion was inferior ST-segment elevation in leads II, III, and aVF (95 percent), and the most common change during LCX occlusion was posterior ST elevation in leads V7, V8, and V9 (68 percent). ST elevation was always seen in inferior leads in the RCA group and in posterior leads in the LCX group. Thus posterior leads helped identify RCA versus LCX as the infarct-related artery. ST elevation was also noted by posterior leads in seven (36.8 percent) additional patients. Thus there was a definite added benefit of posterior leads during LCX occlusion. In the appropriate clinical setting, posterior leads may help in differentiating LCX occlusion from RCA occlusion. PMID- 8721649 TI - Value of proximal regurgitant jet size in tricuspid regurgitation. AB - Recent studies have shown good agreement between proximal regurgitant jet size obtained with transthoracic color flow mapping and regurgitant fraction in patients with mitral regurgitation. To evaluate this in patients with tricuspid regurgitation, we analyzed 40 patients in sinus rhythm, 16 with free jets and 24 with impinging jets, comparing proximal jet size (millimeters) with parameters derived from the Doppler two-dimensional echocardiographic method (regurgitant fraction) and the flow-convergence method (peak flow rate, effective regurgitant orifice area, and momentum). Good agreement was noted between peak flow rate (r = 0.80, p < 0.001), momentum (r = 0.80, p < 0.001), and effective regurgitant orifice area (r = 0.78, p < 0.001), with proximal jet size measured in the apical four-chamber view in patients with free jets. The average of jet proximal size in three planes also had good correlation with peak flow rate (r = 0.75, p < 0.001), regurgitant fraction, momentum, and effective regurgitant orifice area (r = 0.74, p < 0.001). In patients with impinging jets, agreement was fair between effective regurgitant orifice (r = 0.65, p < 0.001), peak flow rate (0.65, p < 0.001), and momentum (r = 0.62, p < 0.001) with mean jet proximal size. Jet proximal size obtained with transthoracic color flow mapping is a good semiquantitative tool for measuring tricuspid regurgitation in free jets that correlates well with established measures of the severity and with new parameters available from analysis of the proximal acceleration field. In patients with eccentrically directed wall jets, the correlation weakens but still appears clinically significant. PMID- 8721650 TI - Systolic anterior motion of the mitral chordae tendineae: prevalence and clinical and Doppler-echocardiographic features. AB - The objective of this investigation was to determine the prevalence and clinical associations of systolic anterior motion (SAM) of the mitral chordae (chordal SAM) in patients without evidence of hypertrophic cardiomyopathy. Although SAM of the mitral valve is thought to be a specific marker of hypertrophic obstructive cardiomyopathy, little is known about the clinical significance of chordal SAM either as an isolated echocardiographic finding (ICSAM) or in patients with mitral valve prolapse (MVP). A retrospective search of the clinical echocardiographic database was made to identify studies demonstrating chordal SAM with no other echocardiographic features of hypertrophic cardiomyopathy. The prevalence of chordal SAM was also ascertained in a group of 97 normal control subjects. Clinical, demographic, and two-dimensional and Doppler echocardiographic characteristics were compared between patients with and without MVP. Chordal SAM was identified in 3.9 percent of clinical studies but was rarely seen in normal volunteers (1 percent). Of the 57 patients with chordal SAM, 21 had systemic or cardiovascular conditions other than MVP associated with SAM (including 7 with aortic insufficiency and 8 with secondary concentric left ventricular hypertrophy), 18 (32 percent) had MVP, and 19 (33 percent) had no associated cardiovascular or systemic condition. These 19 patients with ICSAM were similar to patients with MVP and SAM with respect to age (44 +/- 8 vs 41 +/- 17 years), blood pressure, left ventricular wall thickness, ejection fraction, left atrial size, degree of mitral insufficiency, and left ventricular outflow tract velocity. Indications for the echocardiographic studies were similar between the two groups (chest pain, syncope, arrhythmia, cardiac source of embolus, and suspected MVP), but more patients in the ICSAM group were men (16 of 19 vs 8 of 18; p < 0.05). In conclusion, patients with ICSAM and CSAM associated with MVP are virtually indistinguishable by clinical, demographic, or Doppler echocardiographic features. The syndrome of ICSAM deserves further study as a potentially clinically significant echocardiographic variant of the floppy mitral valve/MVP syndrome. PMID- 8721651 TI - Surgical commissurotomy of the aortic valve: outcome of open valvotomy in neonates with critical aortic stenosis. AB - Early intervention is necessary in neonates with critical aortic stenosis. The advent of alternate therapy, particularly balloon aortic valvuloplasty, requires a reappraisal of the traditional surgical approach, including the efficacy of initial transvalvar gradient reduction and freedom from recurrence of obstruction in the longer term. This report describes a series of 33 consecutive infants who underwent surgical aortic valvotomy in the first month of life. The hospital mortality was 18 percent with a 5-year probability of survival of 66 percent (90 percent CI, 50 percent to 79 percent). Fourteen reinterventions, nine reoperations and five balloon dilatations, were required at a median age of 0.8 years (range 9 days to 6 years). Three patients died after reintervention (one early and two late). The median follow-up time was 5.8 years (range 0.2 to 14 years). At last follow-up Doppler investigation the average Doppler mean and peak gradients were 34 mm Hg and 18 mm Hg, respectively. Open valvotomy in neonatal aortic valve stenosis allows the use of an appropriate surgical approach with low initial mortality and satisfactory 5-year freedom from reintervention. PMID- 8721652 TI - Molecular detection and differentiation of enteroviruses in endomyocardial biopsies and pericardial effusions from dilated cardiomyopathy and myocarditis. AB - Enteroviruses (EVs), especially group B coxsackieviruses, have been implicated in the pathogenesis of myocarditis and dilated cardiomyopathy (DCM). To determine whether a specific type of EV is present in DCM hearts, we examined the genotypes of EVs detected in endomyocardial biopsies and pericardial effusions by polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) analysis. Positive PCR results were obtained from biopsies in 6 (19 percent) of 31 patients with DCM, 5 (18 percent) of 28 with myocarditis, 5 (22 percent) of 23 with other cardiac diseases, and from pericardial effusions in 4 (57 percent) of 7 patients with pericarditis. SSCP profiles of most of the clinical samples were different and were not identical to any of the standard group B coxsackie viruses. Our findings suggest that EV genomes are involved in the myocardium of patients with various cardiac conditions and that a particular type of EV is not present in DCM hearts. PMID- 8721653 TI - Relation of Doppler transmitral flow patterns to functional status in congestive heart failure. AB - We analyzed the relation of Doppler transmitral flow patterns and New York Heart Association (NYHA) classification in 60 consecutive patients with heart failure (44 men and 16 women; mean age 58 +/- 13 years) with left ventricular ejection fraction of < 40 percent. Of the study population, 40 patients with mild to moderate heart failure underwent symptom-limited exercise testing. The relation of transmitral flow pattern and exercise tolerance was also analyzed. Doppler echocardiography was performed in all patients. The patients were subdivided into nonrestrictive and restrictive groups according to the pattern. The univariate analysis showed a relation for left ventricular ejection fraction (p = 0.01 by analysis of variance testing) and transmitral flow pattern (p = 0.0003 by chi squared test) with NYHA classification. When the multivariate regression analysis was performed, only the restrictive pattern by Doppler emerged as an independent determinant of the advanced NYHA class (p = 0.005). In mild to moderate congestive heart failure, patients with nonrestrictive pattern exercised significantly longer than those with the restrictive pattern by an average of 133 seconds (p = 0.003) despite comparable reductions in ejection fraction. Thus the restrictive transmitral flow pattern by Doppler is a noninvasive marker for severe symptoms and diminished exercise tolerance in heart failure patients. PMID- 8721654 TI - Effect of beta-blockade on left atrial contribution to ventricular filling in dogs with moderate heart failure. AB - Abnormal left ventricular (LV) filling has been observed in patients with heart failure. One feature of this abnormality is a reduction in the left atrial (LA) contribution to filling, a feature that can adversely affect overall LV stroke output. In this study we examined the effects of early, long-term monotherapy with the beta-blocker, metoprolol, on LA contribution to ventricular filling in dogs with moderate heart failure. LV dysfunction (ejection fraction 30 percent to 40 percent) was produced in 14 dogs by multiple, sequential intracoronary microembolizations. Dogs were randomized to 3 months' therapy with metoprolol (25 mg twice daily; n = 7) or to no therapy at all (control; n = 7). Mitral inflow velocity was measured before randomization and after completion of therapy by using pulsed Doppler echocardiography. The percentage of LA contribution to LV filling was calculated as the ratio of the time-velocity integral of the LA component of mitral inflow velocity (Ai) to the time-velocity integral of total diastolic inflow velocity (Ti) times 100. In control dogs, the percentage of LA contribution to filling decreased after 3 months of follow-up compared with that before randomization (14 percent +/- 3 percent vs 23 percent +/- 5 percent; p = 0.02). In contrast, in dogs treated with metoprolol, the percentage of LA contribution to filling increased after 3 months of therapy compared with that before randomization (26 percent +/- 3 percent vs 21 percent +/- 2 percent; p = 0.001). Therapy with metoprolol produced a decrease in LV end-diastolic pressure, end-diastolic wall stress and stiffness, and an increase in LA fractional shortening compared with no therapy at all. We conclude that early, long-term therapy with metoprolol improves LA contribution to LV filling. This beneficial effect is likely caused by the ability of beta-blockers to reduce LA workload and consequently improve LA performance. PMID- 8721655 TI - Electrophysiologic characteristics of human atrial muscle in paroxysmal atrial fibrillation. PMID- 8721656 TI - Secular trends in the prevalence of atrial fibrillation: The Framingham Study. PMID- 8721657 TI - Sex-specific differences in coronary artery disease risk factors, evaluation, and treatment: have they been adequately evaluated? PMID- 8721658 TI - Characteristic electrocardiographic features of manifest left anterior paraseptal accessory atrioventricular connection. PMID- 8721659 TI - Mitral ring abscess caused by bacterial endocarditis on a heavily calcified mitral annulus fibrosus: Diagnosis by multiplane transesophageal echocardiography. PMID- 8721660 TI - Rapid growth rate of an apical left ventricular myxoma. PMID- 8721661 TI - Contrast echocardiography as an adjunct in hemorrhagic or complicated pericardiocentesis. PMID- 8721662 TI - Exercise and heart rate variability. PMID- 8721663 TI - Dynamic left ventricular outflow tract obstruction as a complication of acute myocardial infarction. PMID- 8721664 TI - Left main coronary aneurysm and severe three-vessel disease with sparing of an isolated conus artery. PMID- 8721665 TI - Development of a coronary aneurysm 6 months after stent implantation assessed by intracoronary ultrasound. PMID- 8721666 TI - Stenting for coronary dissection after balloon dilation of in-stent restenosis: stenting a previously stented site. PMID- 8721667 TI - Long-term patency of metallic stents in 20-year-old saphenous vein coronary bypass graft. PMID- 8721668 TI - Role of atrial extrastimulation in the diagnosis of atrioventricular node reentrant tachycardia with antegrade atrioventricular conduction via bystander accessory connection. PMID- 8721669 TI - A reappraisal of ganglioside GD3 expression in the CNS. AB - GD3 ganglioside is a major glycolipid component of the developing central nervous system but diminishes considerably as the CNS matures. Despite consistent biochemical data, the cellular localization of GD3 expression has been controversial. In this commentary we will review the cellular expression of GD3 during CNS development and in neuropathological circumstances as determined by studies with the two most commonly used anti GD3 monoclonal antibodies, R24 and LB1. GD3 is not restricted to any one cell lineage, being expressed in development to varying degrees by immature neuroectodermal cells, oligodendrocyte progenitors, ameboid microglia, and subpopulations of developing neurons and astrocytes. In the adult CNS, GD3 is expressed in low amounts by some neuronal subpopulations, on reactive and resting microglia, and by reactive astrocytes. In the appropriate contexts of development or neuropathology, anti-GD3 antibodies are useful for cell type identification and for cell isolation, but caution should be exercised because of the lack of cellular specificity. PMID- 8721670 TI - Modulation of two functionally distinct Ca2+ stores in astrocytes: role of the plasmalemmal Na/Ca exchanger. AB - Mechanisms that regulate the amount of releasable Ca2+ in intracellular stores of cultured mouse astrocytes were investigated using digital imaging of fura-2 loaded cells. At rest, the cytoplasmic Ca2+ concentration, [Ca2+]cyt, was about 110 nM. In the absence of extracellular Ca2+, cyclopiazonic acid (CPA), an inhibitor of the endoplasmic reticulum (ER) Ca(2+)-ATPase, induced a transient, four-fold increase in [Ca2+]cyt due to the release of Ca2+ from inositol triphosphate (IP3) sensitive stores. Caffeine (CAF), which releases Ca2+ from Ca(2+)-sensitive stores, induced a two-fold increase in [Ca2+]cyt. The CPA- and CAF-sensitive stores could be released independently. Changes in the amplitudes of the Ca2+ transients were taken as a measure of changes in store content. Removal of extracellular Na+ or addition of ouabain, which inhibit Ca2+ extrusion and promote Ca2+ entry across the plasmalemma via the Na/Ca exchanger, caused minimal increases in resting [Ca2+]cyt but greatly potentiated both CPA- and CAF induced Ca2+ transients. The amount of Ca2+ releasable from the IP3(CPA) sensitive store was directly proportional to cytosolic Na+ concentration (i.e., inversely proportional to the transmembrane Na+ electrochemical gradient). Under these reduced Na+ gradient conditions, little, if any, Ca2+ destined for the ER stores enters the cells through voltage-dependent Ca2+ channels. These results demonstrate that mouse astrocytes contain two distinct ER Ca2+ stores, the larger, IP3- (CPA-) sensitive, and the smaller, Ca(2+)- (CAF-) sensitive. The Ca2+ content of both ER stores can be regulated by the Na/Ca exchanger. Thus, the magnitude of cellular responses to signals that are mediated by Ca2+ release induced by the two second messengers, IP3 and Ca2+, can be modulated by factors that affect the net transport of Ca2+ across the plasmalemma. PMID- 8721671 TI - Induction of NF-kB-like transcription factors in brain areas susceptible to kainate toxicity. AB - Administration of kainate (KA), a glutamate receptor agonist, to rats causes neuronal damage in the CA1/CA3 fields of the hippocampus and in the pyriform/ entorhinal cortex. Reactive gliosis also occurs and activated astrocytes upregulate their expression of a large number of molecules. Since NF-kB transcription factors are involved in cellular responses to diverse pathogenic stimuli and have been shown to be induced in astrocytes in vitro in response to cytokines and growth factors, we investigated their possible involvement in the changes in gene expression subsequent to KA-induced lesions. Immunoreactivity to the p65 subunit of NF-kB was markedly increased in non-neuronal cells 2 days after KA administration (8 mg/kg i.p.) in the areas of selective neuronal degeneration. This increase was not observed 3 h or 1 day after injection, but was still present 7-10 days after KA injection. By gel mobility-shift assay, a protein complex binding to the kB consensus sequence was found to be induced by 2 days after KA, which correlated with immunohistochemical findings. This NF-kB protein complex seemed to be localized in reactive astrocytes, as indicated by the morphological similarity of NF-kB-positive cells and reactive astrocytes stained with glial fibrillary acidic protein (GFAP) antibody, and the parallelism between the time course of NF-kB induction and appearance of gliosis after KA treatment. Double immunocytochemistry experiments demonstrated the colocalization of NF-kB positive cells and reactive astrocytes. Our results suggest that activated NF-kB in astrocytes participates in delayed and long-term responses of glia to injury. PMID- 8721672 TI - Stimulation of endogenous ganglioside metabolism by neurotrophic growth factors in cultured retinal Muller glia. AB - Neurotrophic factors such as basic fibroblast and epidermal factor (bFGF and EGF respectively) are known to influence many differentiative processes, but their effects on an important group of glycosylated signalling molecules involved in neural differentiation, the gangliosides, are unknown. To study this possibility, we analyzed the effects of exogenously added bFGF and EGF upon the amount and type of endogenous gangliosides extracted from purified cultures of retinal Muller glial cells. A single addition of 500 pM bFGF or EGF for 48 h to such cultures led to significant increases in total ganglioside levels of 30-40%. Analysis of the distribution of specific ganglioside species within control and growth factor treated cells revealed that the precursor form GM3 formed 50-60% of the total ganglioside pool in all cases, the remainder being composed principally of GD1a (20%) with no detectable tri-sialogangliosides. Growth factor treatment for 48 h led to increases mainly in GM3, whereas longer exposure (96 h) of confluent glial cultures to growth factors additionally stimulated synthesis of GT1b. Furthermore, growth factor-induced ganglioside increases were dose dependent, reaching maximal stimulation at 500 pM for bFGF. Incorporation of radiolabelled [3H]-glucosamine into glial cultures showed that ganglioside synthesis was stimulated 2-fold by the growth factors. To our knowledge these data constitute the first demonstration of neurotrophic factor stimulation of ganglioside levels in cells of central nervous system origin. Such complex interactions between peptide growth factors and gangliosides, if occurring in vivo, could have important consequences for retinal cell behaviour. PMID- 8721673 TI - Microglial responses to focal lesions of the rabbit retina: correlation with neural and macroglial reactions. AB - There is a very wide spread Muller glial response to focal laser photocoagulation lesions in the rabbit retina. In this study we have described the microglial response to similar lesions and compared this with the Muller and retinal ganglion cell responses. Microglia were labelled using nucleoside di-phosphatase histochemistry in adult rabbit retinal wholemounts and compared with axonal and Muller cell responses as shown respectively by neurofilament and GFAP immunohistochemistry. In the normal retina, microglia were located in the nerve fibre layer (NFL), inner plexiform layer (IPL), and sparsely in the outer plexiform layer (OPL). Following laser photocoagulation each layer reacted differently. The NFL reaction was exclusively associated with axonal degeneration, as shown by abnormal neurofilament label, and therefore only started several days after injury. In the IPL, neighbouring microglial cells directed their processes towards the lesion by 2 h and had migrated into the lesion by 6 h, but the reaction did not extend more than 2-3 cell diameters from the lesion and was over by 7 days. In the OPL the cell density increased by 1-2 days over a few millimeters from the lesion. The Muller cells expressed GFAP for several millimeters from the lesion starting at 24 h and persisting for over one month and therefore the correlation with the microglial reaction was poor. The different reaction in each retinal layer is evidence that microglial responses are modulated by local factors, probably mainly by contact with injured retinal elements as well as diffusable factors. PMID- 8721674 TI - Endothelin-induced cytoskeletal actin re-organization in cultured astrocytes: inhibition by C3 ADP-ribosyltransferase. AB - We investigated signal transduction mechanisms of endothelin (ET) receptor mediated actin re-organization of rat cultured astrocytes. Staining of filamentous actin (F-actin) showed that stress fibers were a prominent cytoskeletal actin structure in protoplasmic astrocytes. A treatment with 0.5 mM dibutyryl cAMP (DBcAMP) caused cytoplasmic retraction and disappearance of stress fibers of astrocytes. A subsequent addition of 1 nM ET-3 after the DBcAMP treatment expanded the cytoplasm and stimulated stress fiber formation. ET-1, sarafotoxin S6b, and [Ala1,3,11,16]-ET-1 had similar effects. Pre-treatment with 0.1 microgram/ml pertussis toxin (PTX) and chelation of cytosolic Ca2+ did not affect astrocytic stress fiber formation by ET-3. ET-3 stimulated stress fiber formation in stellate astrocytes induced by 50 microM ML-9, 20 microM W-7, and 5 microM cytochalasin B (CB). Cytoplasmic microinjection of C3ADP ribosyltransferase of C. botulinum (C3 enzyme), which impairs the interaction between rho proteins and the effectors, prevented ET-3-induced stress fiber formation and cytoplasmic expansion in DBcAMP-and CB-treated cells. Effects of ET 1 and sarafotoxin on stress fiber formation were also prevented by C3 enzyme. On the other hand, injection of C3 enzyme did not affect increase in cytoplasmic Ca2+ levels induced by ET-3. These results suggest that rho proteins are involved in the ET receptor-mediated actin re-organization of astrocytes. PMID- 8721675 TI - Selective ablation of astrocytes by intracerebral injections of alpha aminoadipate. AB - The efficacy and the specificity of the putative astrotoxin, alpha-aminoadipate, were examined in this study. The integrity of astrocytes was evaluated at several time points following a single injection of alpha-aminoadipate into amygdala of adult rats using immunohistochemistry. The density and the morphological appearance of neurons and the response of microglia were also examined. The injection of alpha-aminoadipate disrupted the astrocytic network in that region. There was a profound loss of glial fibrillary acidic protein-positive and S100 beta-positive astrocytes, normally present in the region, while vimentin immunohistochemistry revealed the presence of deformed cell processes, presumably astrocytic. The presence of reactive microglia at the injection site was suggestive of an active degenerative process, while the normal neuronal density and appearance, as compared to controls, suggested that the damage was confined to astrocytes. The confirmed effectiveness and cellular specificity of alpha aminoadipate in vivo makes it a potentially important experimental tool for attempting to decipher the functional significance of astrocytes. PMID- 8721676 TI - Reactive glial cells express a vitronectin-like protein in the hippocampus of epileptic rats. AB - Injection of kainic acid into the amygdala induces in addition to a local cell loss a seizure related distal damage of the hippocampal complex, in particular in the CA3 field and hilus. This neuronal lesion is associated with hypertrophy and proliferation of astroglial cells which start around 3 days after kainate and peaks within 20 days of kainate. We now report that reactive astrocytes are labelled with antibodies against vitronectin in the CA3 field and hilus. In the present study we also exclude that the presence of vitronectin into the brain is due to an extravasation from serum throughout a blood brain barrier leakage. The present results constitute the first demonstration for a glial expression of vitronectin in vivo. Vitronectin is an extracellular matrix glycoprotein involved in axonal growth. The glial expression of vitronectin may therefore contribute to the synaptic remodeling of mossy fibers induced in the hippocampus by such treatment. PMID- 8721677 TI - Phenotypic diversity and kinetics of proliferating microglia and astrocytes following cortical stab wounds. AB - Brain injury induces reactive gliosis, characterized by increased expression of glial fibrillary acidic protein (GFAP), astrocyte hypertrophy, and hyperplasia of astrocytes and microglia. One hypothesis tested in this study was whether ganglioside GD3+ glial precursor cells would contribute to macroglial proliferation following injury. Adult rats received a cortical stab wound. Proliferating cells were identified by immunostaining for proliferating cell nuclear antigen (PCNA) and by [3H]-thymidine autoradiography, and cell phenotypes by immunocytochemical staining for GD3, GFAP, ED1 (for reactive microglia) and for Bandeiraea Simplicifolia isolectin-B4 binding (all microglia). Animals were labeled with thymidine at 1,2,3, and 4 days postlesion (dpl) and sacrificed at various times thereafter. Proliferating cells of each phenotype were quantified. A dramatic upregulation of GD3 on ramified microglia was seen in the ipsilateral hemisphere by 2 dpl. Proliferating cells consisted of microglia and fewer astrocytes. Microglia proliferated maximally at 2-3 dpl and one third to one half were GD3+. Astrocytes proliferated maximally at 3-4 dpl, and some were also GD3+. Both ramified and ameboid forms of microglia proliferated and by 4 dpl all GD3+ microglia were ED1+ and vice versa. In the contralateral cortex microglia expressed neither GD3 nor ED1. Thus they acquired these antigens when activated. Neither microglia nor astrocytes that were thymidine-labeled at 2, 3, or 4 dpl changed in number in subsequent days. Most thymidine+ astrocytes were large GFAP+ reactive cells that clearly arose from pre-existing astrocytes, not from GD3+ glial precursors. In this model of injury microglia proliferate earlier and to a much greater extent than astrocytes, they can divide when in ramified form, and GD3 is up-regulated in most reactive microglia and in a subset of reactive astrocytes. We also conclude that microglial proliferation precedes proliferation of invading blood-borne macrophages. PMID- 8721678 TI - Retinoic acid regulatory pathways, chromosomal translocations, and acute promyelocytic leukemia. AB - Retinoic acids (RAs) exert a broad range of physiologic actions during embryonic development and adult life. Two families of RA receptors, retinoic acid receptor (RAR) and retinoid X receptor (RXR), have been identified. The therapeutic effect of all-trans-RA (ATRA) in induction of remission for acute promyelocytic leukemia (APL) has largely been proved, and this has, over the past 10 years, greatly stimulated research on oncogenesis and RA-regulated differentiation pathways. In APL, one of the RAR genes, RARA, is fused to PML in the great majority of patients as a result of the chromosomal translocation t(15; 17). However, a small subset of APL patients have a different fusion gene, PLZF-RARA, resulting from the variant translocation t(11;17). A third translocation, t(5;17), in which the NPM gene is fused to RARA, has been described. Current data suggest that PML-RAR alpha and PLZF-RAR alpha fusion receptors may play an important role in the development of APL and that PML-RAR alpha could be the target of ATRA differentiation therapy. Characterization of the genes regulated by retinoic acid may open up new prospects for an understanding of the mechanisms of ATRA differentiation therapy for APL and may help to extend the concept of cancer targeting treatment to other types of leukemias or solid tumors. PMID- 8721680 TI - Genetic mechanisms in esophageal carcinogenesis: frequent deletion of 3p and 17p in premalignant lesions. AB - Although dysplasia of the esophagus is thought to be the precursor lesion of esophageal squamous cell carcinoma (ESC), the sequence of genetic events during esophageal carcinogenesis is unclear. Using the polymerase chain reaction, we examined allelic losses at microsatellite loci in DNAs isolated from 106 lesions among 32 patients with ESC. Allelic losses on 3p or 17p occurred frequently even in dysplastic lesions (9 of 21 and 13 of 24 samples, respectively) including lesions with mild dysplasia (3p, 4 of 10 samples; 17p, 6 of 14 samples, respectively), and allelic losses on these chromosomal arms were also observed in cancerous tissues. We also detected allelic losses of the short and long arms of chromosome 9 at a low frequency in lesions with mild dysplasia and often in lesions with severe dysplasia and in intraepithelial cancers. Our results suggested that inactivation of tumor suppressor genes on 3p and 17p occurs at a very early stage of esophageal carcinogenesis and that genes on 9p and 9q are likely to play important roles in malignant changes. Comparison of the genetic alterations in precancerous dysplastic lesions with those in carcinomas supports the idea that ESC arises from the dysplastic lesion. PMID- 8721679 TI - Sensitive detection of loss of heterozygosity in the TP53 gene in pancreatic adenocarcinoma by fluorescence-based single-strand conformation polymorphism analysis using blunt-end DNA fragments. AB - We have developed a fluorescence-based single-strand conformation polymorphism analysis to detect Haelil-sensitive polymorphic sites in intron 1 of the TP53 gene. It is important to treat the PCR products with Klenow fragment to remove a 3'-protruding nucleotide from the amplified DNA fragments added during the reaction in order to obtain a single peak for each allele. A comparison of the signal profiles of two alleles with those of normal heterozygotes by data processing using computer software has enabled sensitive detection of loss of heterozygosity (LOH) from clinical materials with a fraction of tumor cells below 10%. In analysis of 14 pancreatic carcinomas in which the proportion of the tumor cells is usually low due to the abundance of the stromal component, 7 samples (50%) were informative and 5 of the 7 (71.4 %) were positive for LOH at the TP53 locus. This approach would be useful for allelotyping tumors with low cellularity, as well as other clinical samples such as biopsied specimens and paraffin embedded tissues. PMID- 8721681 TI - Cytogenetic analysis of three primary Bellini duct carcinomas. AB - Three Bellini duct carcinomas (BDC) of the kidney were cytogenetically analyzed after short-term culture. All three had clonal chromosome abnormalities: 91 92,XXY,-Y, +12, +12, -15, -16, -18, +mar (case 1); 53,XY, +2,t(2;7)(p22;q11), +der (2)t (2;7)(p22;q11), +3, +r(3),add(5)(p15), +7, -8, +12, +17, +r(17), +20, 21 (case 2); and 44-47,X,-Y, +9, +16, -21/46,XY. Some of the numerical abnormalities are shared with papillary renal cell carcinomas (PRCC)(+7, +12, +16, +17, and +20) but not with transitional renal cell carcinomas. The present findings support the previous notion that BDC are different from other types of RCC. PMID- 8721682 TI - Molecular variant of the NPM-ALK rearrangement of Ki-1 lymphoma involving a cryptic ALK splice site. AB - The breakpoints of the translocation t(2;5)(p23;q35) associated with Ki-1 positive anaplastic large cell lymphoma (Ki-1 ALCL) have recently been cloned. They involve a novel tyrosine kinase gene, ALK, at 2p23 and the nucleophosmin gene, NPM, at 5q35. Reverse transcriptase-polymerase chain reaction (RT-PCR) with NPM and ALK primers detects a consistent fusion product in Ki-1 ALCL cases that have the translocation. In the course of a survey of 15 cases of Ki-1 ALCL, we identified a single case with a slightly smaller NPM-ALK RT-PCR product, among 12 cases positive for this fusion RNA. Sequencing of this novel NPM-ALK RT-PCR product showed an in-frame junction of NPM to ALK, 30 bases distal to the usual ALK junction site, but at the usual NPM Junction site. The predicted chimeric protein in this case is thus shorter by 10 amino acids, but the putative ALK catalytic domain remains intact. PCR with ALK primers bracketing the novel fusion point, performed on either cDNA or genomic DNA, yielded the same product, confirming that this novel ALK fusion point was located within an exon. Hybridization analysis of the genomic junction fragment isolated by long-range DNA PCR suggested that the ALK genomic breakpoint was also exonic. Cloning and sequencing of the genomic breakpoint confirmed that the break occurred within the 5' portion of the ALK exon participating in the fusion junction, 28 bases 3' to the normal ALK exon boundary, resulting in the use of a cryptic splice acceptor site two bases distal to the breakpoint. This case demonstrates that, in translocations resulting in chimeric transcripts, genomic breakpoints may rarely lie within an exon, provided that the reading frame is maintained and no domains presumed critical to tumorigenesis are deleted. PMID- 8721683 TI - Clonal chromosome abnormalities in two chemodectomas. AB - Short-term cultures from two histologically benign chemodectomas, one from the carotid body and one from the vagal nerve, were analyzed cytogenetically. The former had a small abnormal clone with the karyotype 46,XX,t(3;19)(q21;q13),t(12;15) (p13;q12-14), whereas the majority of the cells from the latter tumor displayed two related abnormal clones: 46,XY,i(I)(q10)/ 46,iderm,add(2)(q37). The findings add to the evidence that chemodectomas are heterogeneous neoplasms and suggest that the heterogeneity may possibly be associated with the site of origin. PMID- 8721684 TI - Loss of the chromosomal region 5q11-q31 in the myeloid cell line HL-60: characterization by comparative genomic hybridization and fluorescence in situ hybridization. AB - Comparative genomic hybridization was used to identify the regions of genomic gain and loss in the myeloid cell line HL-60. These included amplification at 8q24 corresponding to previous reports of overrepresentation of the MYC gene; loss of material from the short arms of chromosomes 9 (9p21-p23), 10, and 17; loss of the chromosome regions 9q32-qter and 14q11-q24; and an extra copy of chromosome 18. Additionally, deletion of the 5q11-q31 region was noted and was associated with translocation of chromosome 5 material to chromosomes 16 and a dic(5;17)(q11;p11) chromosome (previously described as mar 3). Loss of chromosome 5 material in myeloid malignancies, including the M2 subtype from which HL-60 was derived, is usually associated with interstitial deletions of the long arm, including the critical 5q31 region, resulting in a 5q- chromosome. The HL-60 cell line may be a useful model to investigate the role of potential tumour suppressor genes associated with loss of 5q material in myeloid leukaemias. PMID- 8721685 TI - Infrequent mutations of the TP53 gene and no amplification of the MDM2 gene in hepatoblastomas. AB - We have investigated the mutation of the TP53 gene in hepatoblastomas (HBLs) by using polymerase chain reaction-single strand conformation polymorphism and direct sequencing in 38 HBL tumor samples and in two HBL cell lines. We detected the TP53 gene mutation in an anaplastic hepatoblastoma cell line, but no aberration of the TP53 gene (exons 5-9) was found in tumor samples and in the other HBL cell line. The mutation of the cell line was a missense mutation from GAC (asparagine) to CAC (histidine) at codon 281, which was different from the G to-T transversion of codon 249 that is frequently found in adult hepatocellular carcinomas (HCCs). In addition, we performed Southern blot analysis of the MDM2 gene, but we did not find MDM2 gene amplification in 19 cases tested. Our results suggest that, in contrast to the findings in HCCs in adults, TP53 gene aberrations are not involved in the development or progression of HBLs in children. PMID- 8721686 TI - Identification of a YAC spanning the translocation breakpoint t(8;22) associated with acute monocytic leukemia. AB - Using a series of yeast artificial chromosomes (YAC) from the Bp11-12 chromosome region, we have analyzed a t(8;22) translocation present in two patients suffering from acute leukemia by using fluorescence in situ hybridization (FISH). We have identified a YAC that spans the breakpoint in both cases. PMID- 8721687 TI - Have cancer cytogeneticists been wrong in assigning chromosome 12 breakpoints in benign tumors? PMID- 8721688 TI - Elective implantation of intracoronary stents without intravascular ultrasound guidance or subsequent warfarin. AB - Two hundred forty-three stents (203 Palmaz-Schatz, 40 Glanturco-Roubin) were electively Implanted in 188 lesions in 168 patients (mean age 58 +/- 10 years, 77% males) using angiographic but not ultrasound guidance. Patients were treated subsequently with aspirin and observed in hospital for up to 7 days. Those with acute myocardial infarction, radiolucent defects in coronary arteries suggestive of thrombus, and results that were not optimal after stent implantation were anticoagulated with warfarin and not Included in the study. Two had subacute stent thrombosis and two patients non-Q-wave myocardial infarction in-hospital. At follow-up (median 149 days) none had had subacute stent thrombosis, one suffered non-Q-wave myocardial infarction, none had died, and none had developed major complications at the vascular access site. Fourteen (8%) had undergone further revascularisation procedures. This initial experience suggests that aspirin is sufficient to prevent subacute stent thrombosis after elective high pressure assisted coronary stent implantation without intravascular ultrasound guidance if the angiographic appearance after stent deployment is optimal. PMID- 8721689 TI - Do you need IVUS guidance for coronary stent deployment? PMID- 8721690 TI - Femoral artery hemostasis using an implantable device (Angio-Seal) after coronary angioplasty. AB - Coronary catheter interventional procedures are associated with risk of access site complications. We report our experience with Angio-Seal, an implantable hemostasis device, when used in the femoral artery after coronary angioplasty procedures. Sixty-eight patients were studied. Their average age was 63 years; 84% of the patients were male. All had 8 French access sheaths and received bolus heparin (mean dose 12,690 U). The arterial sheaths were removed an average of 455 min after the conclusion of the procedure, when the activated clotting time was 220 +/- 94 sec (range 97-503 sec). The hemostasis device was successfully deployed in 63 patients (93%). The average time to achieve complete arterial hemostasis was 4.4 +/- 8.9 min (range 0-45). Immediate, total hemostasis without requiring any form of external pressure was obtained in 37 of these patients (54%). the incidence of complications was as follows: significant bleeding occurred in 9 patients (13%); there were 2 hematomas (3%); there were no vascular or infectious complications. One device embolization occurred when the connecting suture broke and the intravascular anchor was lost; no clinical sequelae resulted, and manual hemostasis was successful. In four other patients, the device did not deploy and was removed entirely, followed by uneventful manual hemostasis. Follow-up for 2 months revealed no late sequelae in any patient, and complete absorption of the device was documented by ultrasound study in all cases. We conclude that this implantable device can achieve arterial hemostasis quickly and safety when used in anticoagulated patients after coronary interventional procedures. PMID- 8721691 TI - Has the time come for the plug? PMID- 8721692 TI - Initial experience using Prostar: a new device for percutaneous suture-mediated closure of arterial puncture sites. AB - A new device that enables closure of the femoral artery puncture site by percutaneous placement of two nonabsorbable sutures (Prostar) was evaluated. Our initial experience included 32 insertion attempts at 29 femoral arterial puncture sites and one femoral venous puncture site. The device was applied at arterial puncture sites that had been used to carry out 12 balloon angioplasties (41%), seven intracoronary stent placements (24%), five intraaortic balloon pump insertions (17%), four diagnostic angiographies (14%), and one rotational ablation (3%). The venous access site closed was in a patient who had undergone balloon angioplasty and intracoronary thrombolysis. Most patients were anticoagulated with an average activated clotting time (ACT) of 306 +/- 123 sec (12 patients) or an average PTT of 68 +/- 29 sec (14 patients). There were four failures to achieve hemostasis using the device due to: inability to place the device because of peripheral vascular disease, entrapment of cutaneous tissue in the suture, a suture break that prevented hemostasis from being achieved, and avulsion of the sutures from the needles. Although three other suture breaks occurred, these did not prevent hemostasis from being achieved. Thus, 88% (28/32) of attempted uses were successful, and by using a second device in two of the failed attempts, 94% (30/32) of the puncture sites were successfully closed using the device. There was one late rebleed that required 1 hr of groin clamp pressure in an angioplasty patient who had received intracoronary urokinase. An ooze of blood occurred in 4 patients, but in only 2 was this more than trivial, resulting in discontinuation of heparin in one patient and a small hematoma in the other. We conclude that this device can be used safely and effectively, even in fully anticoagulated patients who have undergone complex procedures. The ultimate role of the device will require further experience and appropriate randomized studies. PMID- 8721693 TI - Percutaneous hemostatic closure systems: is the cost worth the price? PMID- 8721694 TI - Slow coronary flow: clinical and histopathological features in patients with otherwise normal epicardial coronary arteries. AB - Slow flow of dye in epicardial coronary arteries is not an infrequent finding in patients during routine coronary angiography. Whether this pattern of flow can be reversed by nitroglycerin or dipyridamole and whether this angiographic finding is associated with histopathological abnormalities is unknown. We hypothesized that this abnormality could be associated with small vessel disease of the heart, since the epicardial arteries are usually widely patent. Thus, out of the patients undergoing heart catheterization at our institution during the past 5 years, 10 (7%) presented with chest pain, normal epicardial coronary arteries, and abnormal coronary progression of dye. Rest electrocardiogram (ECG), exercise test, echocardiographic examination, and left ventricular angiogram were normal. Coronary angiography showed slow flow of dye on a total of 20 main coronary vessels, that was not reversed by intracoronary nitroglycerin administration. Six of them underwent dipyridamole intravenous infusion that normalized dye run-off in all affected vessels, for a total of 9 main coronary vessels. Histopathological examination (light and electron microscope) of left ventricular endomyocardial biopsies showed thickening of vessel walls with luminal size reduction, mitochondrial abnormalities, and glycogen content reduction. Normal and pathological zones often coexisted in the same specimen. Thus. In some patients with slow coronary flow and patent coronary arteries, functional obstruction of microvessels seems to be implicated, as it is relieved by dipyridamole infusion. Patchy histopathological abnormalities suggestive of small vessel disease are also detectable and could contribute to increase flow resistance. PMID- 8721695 TI - Lesion-directed administration of alteplase with intracoronary heparin in patients with unstable angina and coronary thrombus undergoing angioplasty. AB - Percutaneous coronary revascularization in patients with unstable angina and coronary thrombus carries a high complication rate. A new strategy to reduce thrombus burden before revascularization was tested in a multicenter prospective trial. Patients with unstable angina and coronary thrombus (n = 45) received alteplase through an infusion catheter at the proximal aspect of the target lesion and concomitant intracoronary heparin via a standard guiding catheter. Angiography was performed before and alter lesion-directed therapy and post intervention. Systemic fibrinogen depletion and thrombin activation were not observed, while fibrinolysis was evident for > or = 4 hr after treatment. Target lesion stenosis did not change significantly after lesion-directed therapy, but thrombus score was reduced, particularly among patients who had large thrombi (mean 2.2 vs. 1.6, P = 0.02). Revascularization was successful in 89% of patients. Median final stenosis was 30% and mean final thrombus score was 0.4. Complications included recurrent ischemia (11%), MI (7%), abrupt closure (7%), severe bleeding (4%), and repeat emergency angioplasty (2%). Patients with overt thrombus appeared to derive the most angiographic benefit from lesion-directed alteplase plus intracoronary heparin. Later revascularization was highly successful. This strategy may be a useful adjunct to percutaneous revascularization for patients with unstable angina and frank intracoronary thrombus. PMID- 8721696 TI - Core curriculum for the training of adult invasive cardiologists: report of the Society for Cardiac Angiography and Interventions Committee on Training Standards. PMID- 8721697 TI - Core curriculum for the training of pediatric invasive/interventional cardiologists: report of the Society for Cardiac Angiography and Interventions Committee on Pediatric Cardiology Training Standards. AB - Within the field of pediatric cardiology, a number of subspecialty fields are generally recognized. Some of these overlap. For example, most electrophysiologists also would consider themselves to be clinical cardiologists. Some fields, however, are relatively mutually exclusive. For example, most clinical pediatric cardiologists would not consider themselves to be electrophysiologists and would admit that there is a different knowledge, skill, and experience base that separates an electrophysiologist from other specialists within the broader field of pediatric cardiology. Likewise, it is our opinion that a separate knowledge, skill, and experience base exists among pediatric Invasive/Interventional cardiologists. The purpose of this report is to define the unique knowledge and skill base required for the training of an invasive pediatric cardiologist. The scope of this report is limited to the training of Invasive/Interventional cardiologists dealing with the treatment of pediatric patients and the cardiac diseases most often encountered in these patients. This report describes in detail the core curriculum suggested for the training of an invasive pediatric cardiologist. For these purposes, invasive pediatric cardiology encompasses all aspects of pediatric diagnostic cardiac catheterization, whether congenital or acquired. Additional curriculum regarding training for therapeutic or interventional procedures is also addressed because of today's needs: most pediatric cardiac catheterizations are performed to acquire specific data that cannot be obtained otherwise by non-invasive technologies and are required for the best medical or surgical management. However, more and more frequently the need for a cardiac catheterization entertains the possibility of having to proceed with an interventional procedure, and therefore, to conserve the vessels of pediatric patients as well as to consider costs, any invasive pediatric cardiologist should be well trained in most of the accepted interventional pediatric cardiology procedures. The curricula is divided here into five major sections followed by a bibliography keyed to those sections. Also included is a suggested format for the objective evaluation and documentation of the progress of invasive pediatric cardiology fellows. It is intended to complement the core curriculum and provide a means for standardizing the evaluation of invasive pediatric cardiology fellows. PMID- 8721698 TI - Enhanced manipulation of directional coronary atherectomy and coronary angioplasty guide catheters. AB - Manipulation of thin-walled coronary guiding catheters, especially directional coronary atherectomy guides, may sometimes be difficult. This is particularly so in patients with tortuous or ectatic aorto-iliac arterial systems. To improve torque transmission and eliminate kinking, the introducer catheter or a smaller caliber diagnostic catheter is advanced to within 1.5 cm of the distal guide tip. Connecting the rotating hemostatic valve to the introducer hub or diagnostic catheter proximally allows distal contrast injections and pressure monitoring via this inner catheter. Other methods of enhancing guide catheter manipulation are also discussed. PMID- 8721700 TI - Maximizing your guide-given talents. PMID- 8721699 TI - Contrast cine left ventriculography: comparison of two pigtail catheter shapes and analysis of factors determining the final quality. AB - Ventricular arrhythmias or inadequate opacification of the ventricular cavity during contrast-cine left ventriculography frequently interfere with evaluation of regional wall motion, ejection fraction or mitral regurgitation. In this prospective, randomized study traditional (straight) pigtail catheter was compared with a new, large loop pigtail catheter (both 6 French, large lumen) in terms of the quality of the cine left ventriculograms. Straight (Group I) and curved pigtail (Group II) groups were further subdivided randomly into a preset (13 cc per second for 3 seconds with a pressure rise time of 0.5 seconds) injection rate (Group IA, n = 46, and Group IIA, n = 48) or operator definded injection rate (Group IB, n = 49, and Group IIB, n = 45) subgroups. The ventricular tachycardia and couplets occurred at similar frequency among the groups. The curved pigtail subgroups showed significantly more frequent catheter induced mitral regurgitation. However, the opacification and overall quality of the left ventriculograms were distributed similarly between the groups. Because the catheter type and injection protocol did not affect the left ventriculogram quality in our study, variables determining opacification and overall quality rating are analyzed in the overall group of 190 patients. Left ventricular opacification was excellent in 72, acceptable in 108 and marginal in 10 patients. The patients with marginal opacification were significantly heavier (P = .004) with larger left ventricular enddiastolic volumes (P = .019), and smaller amount of contrast volume per enddiastolic volume (P = .005) and kilogram body weight (P = .003). The overall quality of left ventriculograms were excellent in 38, acceptable in 133, and marginal in 19 patients. The patients with excellent left ventriculograms were significantly younger (P = .019) and slightly less heavy (P = 0.09). Significantly more female patients were also in this group (P = .036). Ventricular tachycardia was the most common cause of unsatisfactory left ventriculograms. In the RAO view, deeper (more apical) placement of the catheter was associated with higher incidence of ventricular tachycardia (53%). The most "silent" area was the posterobasal area. In conclusion, the perfect left ventriculogram remains to be an elusive goal in routine clinical practice. When using 6F highflow pigtail catheters and nonionic contrast agents, more basal catheter position and higher contrast volume increase the quality of the left ventriculograms. PMID- 8721701 TI - Role of oximeter error in the diagnosis of shunts. AB - We have analyzed the role that instrument error plays in the oximetric diagnosis of shunts. First, we review the accuracy specifications of instruments currently available for measuring oxyhemoglobin saturation during cardiac catheterization. Then we derive a mathematical model of a left-to-right shunt and illustrate the relationships between the magnitude of shunts and the saturation step-ups they generate. Using the shunt model and statistical methods, we show how oximeter error affects the likelihood of success in detecting shunts of a given magnitude. By computing the probabilities of false negative and false positive diagnoses, the model can be used to determine how the likelihood of successful shunt detection is influenced by factors such as shunt flow, the minimum saturation step-up considered indicative of a shunt, oximeter error, hemoglobin concentration, and the number of blood samples. A spreadsheet is included for those who wish to analyze the diagnostic capabilities of their own instrumentation and clinical methods. Conclusions. (1) Modern instruments for use during cardiac catheterization analyze blood samples with a measurement error ranging from 2.5 to 1% saturation or better. (2) Until more information is available regarding the accuracy of such devices in measuring oxygen content, the detection of shunts should be made on the basis of step-ups in saturation rather than oxygen content (even though step-ups in oxygen content have the potential advantage of being independent of the oxygen-carrying capacity of blood). (3) Averaging the analyses of two or more blood samples from each site and using an oximeter with 1% error greatly improve the probability of reaching the correct diagnosis. (4) Oximeter error of 2.5% or greater is unacceptable by today's standards. (5) With the instrumentation currently available, oximetry can potentially detect saturation step-ups as small as 3.6% and still keep both false negative and false positive diagnoses below 10% if venous blood is adequately mixed. PMID- 8721702 TI - Efficacy of marker wire for intracoronary stenting. AB - The Marker Wire was used for Palmaz-Schatz coronary stent implantation. The Marker Wire is useful in estimating lesion length and in determining the number of stents required, in addition to facilitating stent positioning. PMID- 8721703 TI - Whoops, I didn't realize it was so long! PMID- 8721704 TI - Maintenance of coronary guidewire position during guide catheter exchange. AB - Maintaining the position of a guidewire across coronary artery lesions during angioplasty is important to permit rapid and reliable access. This article describes a technique which enables a guide catheter to be replaced while maintaining coronary guidewire position by using an additional, larger guidewire for support to prevent dislodgment of the coronary guidewire. PMID- 8721705 TI - Left ventriculography revisited. PMID- 8721706 TI - Versatility of the microvena snare in pediatric interventional cardiac catheterization. AB - Improved catheter and device technology has allowed the practice of pediatric interventional catheterization to expand in recent years. One such example of technologic advancement has been the development of an intravascular snare by Microvena (White Bear Lake, MN). We present three patients in which the snare was effectively used to perform definitive transcatheter therapy and avoid the need of more invasive surgical treatment. We believe that the snare device significantly decreased our fluoroscopic and procedural time and facilitated the effectiveness of the specific procedure. PMID- 8721707 TI - Rotational atherectomy with a new device: initial clinical experience. AB - The Bard Atherectomy Catheter is a new rotational atherectomy device that consists of a flexible, hollow, thin-walled cutting catheter that, while rotated at 1,500 revolutions per minute, is advanced across the lesion over a special spiral guidewire system. We report the initial clinical experience with this device in 20 peripheral lesions in ten patients. The majority of patients were treated for limb salvage. All lesions were successfully intervened on by atherectomy followed by adjunctive balloon angioplasty. A reduction to less than 50% stenosis was achieved in 13 of the 20 lesions (65%) after atherectomy but in all 20 lesions (100%) after adjunctive angioplasty for all lesions and stenting for dissections in two. Baseline minimal lesion lumen diameter was 0.8 +/- 0.7 mm with a reference vessel diameter of 4.2 +/- 1.7 mm (75 +/- 21% stenosis). The lumen improved to 2.0 +/- 0.8 mm (45 +/- 19% stenosis) (P < 0.001) following atherectomy and to 3.9 +/- 1.9 mm (13 +/- 16% stenosis) (P < 0.001) after adjunctive angioplasty. The average weight of removed atheroma was 45 +/- 58 mg. All ten patients had initial improvement in symptoms. At 6 months follow-up there was persistent improvement in eight patients and two subjects had undergone amputations. Our early clinical experience with this low profile, flexible atherectomy device, that enables extraction of a large amount of atheroma, suggests that it will become a valuable addition to current atherectomy technologies in small- and medium-sized vessels. The value of this device in coronary vessels is under investigation. PMID- 8721708 TI - Women who lead. AB - This essay reviews aspects of historical and cultural changes that now permit women increasing opportunities to lead both women and men. Women assuming leadership responsibilities undergo psychic reorganization, reworking their personal histories and their modes of interaction. The author challenges women who inhibit their leadership potential to scrutinize their attitudes and to consider the implications for the next generation of women. PMID- 8721709 TI - The vigor of metaphor in clinical practice. PMID- 8721710 TI - The Lacanian subject and grotesque desires: between oedipal violation and narcissistic closure. PMID- 8721711 TI - Freud's Acropolis revisited. PMID- 8721713 TI - Unconsciously induced song recall: the process of unintentional rather than so called spontaneous evocations of music. AB - This paper has presented the basis for a reformulation of the traditional psychoanalytic usage of the term spontaneous evocations. Illustrated with a supervisory example, I have argued for a return to Freud's original use of the word unintentional or to equivalent synonyms. The process whereby such unexpected music evocations appear in consciousness has been termed "unconsciously induced song recall." The role of serendipity in this theoretical reformulation will be addressed separately (see Diaz de Chumaceiro, 1995; Diaz de Chumaceiro and Yaber, 1994). Like dreams, music evocations can be subjected to analysis and interpretation, if when recalled by indirect or direct induction in treatment they are mentioned to the therapist. This basic principle of induced recall can be generalized to evocations of the art world at large. PMID- 8721712 TI - Anne Tyler: the American family fights for its half-life. PMID- 8721714 TI - Voice of the analyst. PMID- 8721715 TI - Homosexuality. PMID- 8721716 TI - Homosexuality. PMID- 8721717 TI - Consultancy to troubled therapies. PMID- 8721718 TI - Consultancy to troubled therapies. PMID- 8721719 TI - Consultancy to troubled therapies. PMID- 8721720 TI - Response to reviews of Karen Horney: a psychoanalyst's search for self understanding. PMID- 8721721 TI - [The use of hearing aids in children with profound deafness. Methodology and results]. AB - The early application of hearing aids in children with severe and profound hearing loss is rather an hard problem that has to be solved, according to the Authors, by taking care of several factors. Usually, as far as these patients concern, two kinds of hearings aids are used. The first one emphasizes all the frequencies in the same way, including low tones that, in the early ages, are very important for improving prosodic pattern, occlusive and liquid phonemas, primary words and sentences. The second type, emphasizing only medium and high frequencies, is useful for discrimination processes. The Authors suggest to applicate the hearing aids in these patients, following a behavior be themselves named "dynamic application". In the first phase the Authors use hearing aids emphasizing also low tones; in the second one they modify the device, according more precise data turning out form audiometric evaluation and speech therapy. The results concerning a research on 27 children with profound hearing loss are debated; the application of hearing aids occurred in early age and the speech therapy followed the same criteria for all patients. The subjects were divided in two groups, homogeneous for age, degree of hearing loss and social environment. For the 12 children of groups A the "dynamic application" was used. For the 15 patients of group B the traditional strategy, using only devices for the amplification of medium and high tones, was applied. The best results concerning voice intonation and learning occlusive phonemas were obtained in group A subjects. The Authors, introducing their results and discussion, compare them with references data. In conclusion, the "dynamic application" is effective because of the better results in speech therapy and also because its ratio permits the application of hearing aids in children with hearing loss of different degrees, especially in the most difficult cases. PMID- 8721722 TI - [Nasal polyps: comparative immunological study of polyps with different histopathologic types]. AB - Cellular Infiltrate as well as class I and II HLA molecule expression, on 22 nasal polyps and on 12 samples of corresponding hypsilateral mucous membrane were studied by means of immuno-histological methods. These nasal polyps were classified according to their histopathological structure. Five polyps, with a fibrous connective core infiltrated by cells of the monocyte-macrophage lineage, were classified mixed. The remaining seventeen polyps were characterized by the presence of central oedematous connective tissue infiltrated almost exclusively by eosinophils and either contained (glandular type) or did not contain (oedematous type) glands. A comparative study of different types of nasal polyps and corresponding hypsilateral nasal mucous membranes was carried out on atopic and non-atopic patients. No correlation between atopic status and polyp presence or polyp typology was found. On the other hand, different polyp types appear to have a structural correlation with the corresponding hypsilateral mucous membrane regarding infiltrate cell type, oedematous or fibrous connective tissue presence and expression of on HLA antigen positivity pattern. The characteristic histological structure of hypsilateral mucous membranes in patients with different types of polyps appeared to be brought about by a multifactorial etiology involving mucosal hyperreactivity. Lastly, both polyps and parapolypal nasal mucous membranes were found to be infiltrated mainly in the peripheral subepithelial connective tissue by lymphocytes (55%) as well as by other leukocyte types. The presence of growth factors capable of enhancing an increase of fibroblasts, endothelial cells, together with focal distrupture on the basal membrane, might well be a general mechanism responsible for polyp sprouting. PMID- 8721723 TI - [Nasal patency patterns observed during working hours in a group of technicians habitually exposed to formaldehyde]. AB - Formaldehyde is a substance which is widely used in several work fields, including many medical and biological sectors. It appears that formaldehyde exposure may cause bronchial asthma. Little information is available, however, with regard to upper aereal tracts. The nose is a target organ of major importance for a large range of professional and extra-professional environmental pollutants. In particular, cases of professional rhinitis brought about by exposure to disinfectants have been reported in Literature. In the light of general knowledge related to the close correlation between upper and lower airways, and to the lack of information concerning the effects of short-term formaldehyde exposure on the upper aereal tract, the Authors decided that investigation of such exposure might prove interesting as well as worthwhile. A group of fifteen technicians who work in three different pathology departments and who had been exposed to formaldehyde for at least one year were studied. The purpose of the study was to assess possible alteration in nasal resistances at the end of a normal work shift. The subjects under study were observed according to a procedure which included several clinical, allergological, serological and functional evaluations carried out on specific days of the week. A control group, consisting of eight asymptomatic, non-atopic subjects who had never been exposed to inhalation of irritants was also monitored in the same way. Conclusions were drawn after considering some important variables such as exposure time, possible anatomical alterations of the nose, the presence of respiratory allergies and smoking habits. It was apparent that professional formaldehyde exposure typical of that experienced by technicians working in pathology departments induced a short-term irritant effect on the upper aereal tract, which, even though subclinical, is revealed by a statistically significant increase in nasal resistance. This effect seems to be favoured (at statistically not significant levels) by atopy, cigarette smoking and anatomical alterations of the nose. Our data seem to exclude a chronic cumulative effect of formaldehyde exposure. PMID- 8721724 TI - [Combined transfacial and infratemporal approaches for T3-T4 malignant maxillary tumors]. AB - The technique of maxillectomy employing an anterior transfacial approach has practically remained unaltered since historical and suggestive descriptions of Lizars reported in 1829 and those of Fergusson reported in 1842. This procedure is suitable and efficacious for benign tumors as well as for malignant tumors which involve the inferior, anterior, medial or lateral wall of the maxillary sinus. However, when the tumor erodes the posterior wall, surrounding and destroying the pterygoid and invading the pterygo-maxillary and the infratemporal fossae, sometimes causing thrismus as a result of infiltration of the pterygoid muscles, surgical control of the posterior extension through an anterior approach is impossible. Many authors maintain that in these cases surgery is useless if not detrimental in light of the low survival rates reported. We propose a new surgical technique (a double approach--transfacial and infratemporal preauricolar) to be followed in these patients. These approaches allow us to completely surround the extension of the tumor as well as to achieve surgical radicality in T4. Moreover, with this technique it is possible to use the temporalis muscle to repair the resulting post-maxillectomy cavity thus eliminating the necessity of the obturator. From 1992 to 1994 we operated 46 patients with T3 and T4 malignant tumors of the maxillary sinus following this procedure. We had 1 death due to heart infarction 3 days after surgery. On the other hand no serious local complications were observed. There were only 7 suppurations in the temporal region, either resolved quickly and spontaneously or through simple surgical drainage. The follow-up is still too brief to allow us to draw definite conclusions. At any rate, presently 30 of our 46 patients are alive and disease free. PMID- 8721725 TI - [Widened forwarding total laryngectomy ("squared laryngectomy"). Hints of surgical techniques and personal experience]. AB - Primitive T4 laryngeal neoplasms with anterior invasion and neoplasm recurring after partial and subtotal intervention often invade the soft prelaryngeal tissues and in these cases the neoplastic illness can be no longer be controlled be "organ surgery". The widened forwarding total laryngectomy, "squared" or "carre" laryngectomy according to some Authors of French School, is a surgical procedure not "on an organ" but "in an area" or "region" which proposes to delete, in one step, the larynx, the bone hyoid, the fasciae and the prelaryngeal muscles, the thyroid gland and, if necessary, a more or less large quantity of anterior cervical skin. If the removal involves a vast cutaneous area, it is necessary to mend the loss of substance by wrapping around a miocutaneous flap of pectoralis mayor muscle. In the last five years, 4 male patients, between 48 and 73 years, were treated with widened forwarding total laryngectomy. They were all carriers of epidermoid laryngeal carcinomas with various degrees of differentation: primitive in one patients, recidivist after performance of partial (cordectomy) and subtotal (two Labayle) surgery in the other three patients. In the only case of T4 primitive laryngeal neoplasm it was necessary to carry out a functional neck dissection bilaterally. Loss of substance always required the use of a miocutaneous flap of pectoralis mayor muscle except in one patient in which the removal of the prelaryngeal tissues was limited and therefore it was possible to make a direct seam. We always completely removed the thyroid gland, the prelaryngeal muscular system and skin of the preceding stomy (in the Labayle) sparing, on the other hand, the hyoid bone. Only one patient, who died due to recurrence a year after surgery, underwent complemental percutaneous radiotherapy. At present, three patients are alive and NED: one after 5 years, the others are in excellent conditions although the follow-up is still brief. According to our experience, we can affirm that in selected cases, after an accurate general evaluation of the patient (exclusion of distant metastases, preparation from a metabolic and psychological point of view) a widened forwarding total laryngectomy is a valid procedure since surgery (together with other complementary therapies), is still today the best treatment in forms with anterior evolution. PMID- 8721726 TI - [The treatment of benign laryngeal pathologies using CO2 laser in direct microlaryngoscopy with propofol-induced endovenous anesthesia in spontaneous respiration without intubation]. AB - Direct microlaryngoscopy is an endoscopic technique with considerable diagnostic and surgical potentialities, which are increased when carbon-dioxide Laser is used, but often limited by the anaesthesiologic methods employed. In fact, the traditional anaesthesiologic technique, which uses small bore oral-tracheal tubes and provides good ventilation, has significant disadvantages: the tube often impedes surgical activity especially in the posterior regions of the larynx; stenoses of the larynx impede oral-thacheal intubation and thus require pre operatory thacheotomy; administration of succinylcholine often induces diffused myalgia in the following 12-24 hours; administration of traditional narcotics always requires hospitalization. Jet-ventilation used by the Freach School as an alterative, while offering a more ample operatory space, is also accompanied by many disadvantages: vocal cords vibration; spray expulsion of smoke, blood, etc.; as with the traditional method, it cannot be employed in cases of laryngeal stenoses; succinylcholine often induces diffused myalgia; it cannot be used on out-patients. Introduction of Propofol, a new endovenous anaesthetic, the anaesthetologic pratice has permitted total endovenous anaesthesia to be achieved, in spontaneous respiration, without intubation. This method is indicated even in cases where the previously mentioned approaches are shown to be inadequate or impracticable. Narcosis with Propofol was used in 70 patients with benign laryngeal pathologies, belonging to the ASA risk classes I-II who underwent surgery via CO2 Laser during Microlaryngoscopy. The pharmacologic protocol of this narcosis require intravenous premedication with atropine and phentanyl, induction and continuation with Propofol and pharyngolaryngeal local anaesthesia with lidocaine spray. The advantages offered by this method are considerable: a completely free operative space; the possibility of being used in cases of laryngeal stenosis; a rapid return to consciousness; the absence of myalgia after surgery; the possibility of treating the disease on an out-patient basis. The absence of significant complications and the good results obtained lead us to propose this anaesthesiologic method as a valid alternative to anaesthesia by the traditional oral-tracheal intubation and to Jet ventilation. PMID- 8721727 TI - [Non-Hodgkin's lymphomas of the salivary gland: analysis of prognostic factors in 28 cases]. AB - Primary malignant lymphomas of the major salivary glands are rare and usually arise in the parotid gland (2% of all neoplastic disorders). In this report clinical records of 28 cases of NHL of salivary glands (27 in the parotid gland and one in the submandibular gland) are reviewed and problems related to diagnosis and management strategies are discussed. The 5-year overall survival rate was 72% and did non differ from the survival of other NHL of the head and neck. Statistical evaluation of prognostic factors (age, histology, clinical stage, grading, bulky and surgical approach--biopsy versus parotidectomy), are presented. Analysis of these factors showed that prognosis was not influenced by age, histology, clinical stage and grading of disease. Poor survival was significantly correlated to bulky lesions (tumor size greater than 6 cm). In our experience surgical treatment did not significantly affect survival rate. It is concluded that diagnostic surgical procedures in case of suspected NHL of the parotid gland are fine needle aspiration biopsy. (FNAB) or incisional biopsy. The treatment of choice is radiotherapy associated with chemiotherapy in cases of localized-bulky or disseminated disease. PMID- 8721728 TI - [Logopedic rehabilitation of laryngeal granulomas]. AB - Posterior laryngeal granuloma is an infrequent pathology of multidisciplinary interest. Actually, its real prevalence is difficult to quantify because in some cases it is asymptomatic and in other instances it may either be reabsorbed or eliminated spontaneously. It is located at the vocal apophysis of the arytenoid or, less frequently, above it or on the laryngeal side of the arytenoid. The many etiologic factors (laryngeal intubation, gastro-esophageal refluxes, blunt trauma of the larynx, vocal dysfunction), sometimes concomitant and with the possible addition of enhancing circumstances (upper aerodigestive tract inflammation, naso gastric tube, smoking and alcohol abuse), converge to a single pathogenetic mechanism: an ulceration of the mucosa and the pericondrium, sometimes complicated by an infection, which does not heal but instead produces a typical granulation tissue with capillaries oriented radially from the center of the lesion. Post intubation granulomas, extremely rare in children, are more frequent in females. It appears that there is no correlation with duration of intubation in that granulomas, can also occur after short general anesthesia. Idiopathic or contact granulomas are more frequent in the males. They are the result of vocal laryngeal hyperfunction, habitual throat clearing or cough-like throat clearing. Gastro-esophageal reflux of gastric juice, coughing or throat clearing may injure the mucosa. A blunt trauma of the larynx may cause a granuloma if the cartilage of the vocal process is exposed. Symptoms, when present, are dysphonia, tiredness during or after voicing, bolus, laryngeal unilateral pain, sensation of something in the throat which is mobile during breathing and swallowing, traces of blood in the expectoration. Therapeutic options are surgical, medical or logopedic. Surgery, although followed by frequent recurrences, is mandatory when the granuloma causes dispnea or if a pathologic essay is needed. Medical treatment aims at solving gastroesophageal reflux and/or inflammations of the district. Logopedic rehabilitation is the most successful therapy. Since January 1992 the Authors have been adopting the rehabilitation protocol planned by the French phoniatrician Brigitte Arnoux-Sindt for post-intubation granulomas, which, moreover, is utilyzed for all type of granulomas, including those arising during the early postoperative period after cordectomy. This protocol is analytically presented and discussed. In the cases of contact granulomas, and when there is concomitant vocal dysfunction, logopedic treatment is prolonged after granuloma dissapearance with some sessions aiming at restoring correct vocal behaviour. In all the ten patients rehabilitated up to now, granulomas disappeared after a mean of 16.3 sessions held twice a week. After several months of follow-up we had no recurrences. This clinical experience, while limited in number, seems to confirm the good results already reported in French Literature. PMID- 8721729 TI - [Speech perception test in Italian language for profoundly deaf children]. AB - Speech perception tests are an important part of procedures for diagnosing pre verbal hearing loss. Merely establishing a child's hearing threshold with and without a hearing aid is not sufficient to ensure an adequate evaluation with a view to selecting cases suitable for cochlear implants because it fails to indicate the real benefit obtained from using a conventional hearing aid reliably. Speech perception tests have proved useful not only for patient selection, but also for subsequent evaluation of the efficacy of new hearing aids, such as tactile devices and cochlear implants. In clinical practice, the tests most commonly adopted with small children are: The Auditory Comprehension Test (ACT), Discrimination after Training (DAT), Monosyllable, Trochee, Spondee tests (MTS), Glendonald Auditory Screening Priocedure (GASP), Early Speech Perception Test (ESP), Rather than considering specific results achieved in individual cases, reference is generally made to the four speech perception classes proposed by Moog and Geers of the CID of St. Louis. The purpose of this classification, made on the results obtained with suitably differentiated tests according to the child's age and language ability, is to detect differences in perception of a spoken message in ideal listening conditions. To date, no italian language speech perception test has been designed to establish the assessment of speech perception level in children with profound hearing impairment. We attempted, therefore, to adapt the existing English tests to the Italian language taking into consideration the differences between the two languages. Our attention focused on the ESP test since it can be applied to even very small children (2 years old). The ESP is proposed in a standard version for hearing impaired children over the age of 6 years and in a simplified version for younger children. The rationale we used for selecting Italian words reflect the rationale established for the original version, but the choice of single words follows different criteria from the original version. In fact, the two languages differ in important linguistic features so that the test can not be not adapted to the Italian language by simply translating the words involved. As currently there is no children's language dictionary in Italian arranged according to age bracket, we chose words used in children and in pre-school reading material. PMID- 8721730 TI - [Position of semicircular canals in humans and cryptogenetic cupulolithiasis]. AB - Semicircular canals in humans are three membranous tubes each forming two-thirds of a circle, arranged in a coordinate system. The horizontal semicircular canal makes a 30 degree angle with the horizontal plane, a phenomenon neither easily understood nor explained in Literature. This paper reports a personal opinion based on the theory of evolution. Before man assumed erect posture (Australopithecus Afarensis), more than 3 million years ago, the head was bent forward (as in other primates) so that the temporal bone (structures inside included) laid on the horizontal plane. In the Author's opinion the present alignment might have come about after the gradual change in posture in human evolution. The ampulla of the posterior canal and the non ampullary branch of the horizontal semicircular canal became the lowest part of the utricule thereby allowing otoconial debris to settle. On the basis of this hypothesis the Authors attempt to explain the high prevalence of idiopathic cupulolithiasis. PMID- 8721731 TI - [The history corner: economic sanctions of 1935 and relations between Italian and French otolaryngologists]. PMID- 8721732 TI - [Comments on organization and activity of ENT day hospital department: three years of experience]. AB - The authors deal with organizational and technical aspects concerning otorhinolaryngological day hospital's management. They explain purposes and reasons leading to the establishment of this sort of service; they analyse related problems and the adopted methodologies for simplifying admission procedures. They report a statistical survey regarding the period from May 1991 to May 1994, concerning surgical and medical operations, and instrumental diagnostic researches performed at the IV ENT Department of "La Sapienza" University of Rome. The statistical analysis concerning the 1526 patients sent to this department, shows the downright prevalence of surgical operations performed in operating room (52%). Ambulatory operations and instrumental diagnostic researches account respectively for 39.6% and for 29.3% of all hospitalizations. Among all surgery operations performed in operating room stand out the notice incidence of therapeutical and diagnostic microlaryngoscopies. The most frequent ambulatory operations were the removal of tracheal cannula (21%), while TC was the most frequent of all instrumental diagnostic researches. A careful selection of patients, on the basis of their general condiction, account for the low incidence of subjects not discharged within 24 hours. After having analyzed the survey's data, the authors underline advantages and finalities of day hospital: a shorter hospital stay, reduction of hospitalization's costs, and a larger related number of patients to be contented. Finally they hope that this sort of service will spread more. PMID- 8721733 TI - [letter]. PMID- 8721734 TI - Improved protocol for the formation of N-(p-nitrobenzyloxy)aminoalditol derivatives of oligosaccharides. AB - An improved procedure has been developed for the rapid derivatization of oligosaccharides with UV-detectable p-nitrobenzylhydroxylamine (PNB). The improved conditions used result in quantitative derivatization of neutral oligosaccharides. Sialylated oligosaccharides can also be quantitatively PNB derivatized without detectable desialylation. Of the oligosaccharides tested, only the derivatization of oligogalactosyluronic acids was incomplete (yield approximately 70%). PNB-derivatization of tamarind seed xyloglucan oligosaccharides results in products with improved chromatographic properties during HPAEC. These PNB derivatives were also subjected to hydrophilic interaction chromatography (HILIC) and analyzed by on-line LC-MS. On-line LC-MS is readily usable with HILIC, as this chromatographic technique does not require salt-containing solvents. Approximately 10 pmol of a PNB-derivatized oligosaccharide can be identified and quantitated utilizing this method. PMID- 8721735 TI - Synthesis and structural characterization of 6I,6II-diamino-6I,6II-dideoxy cyclomaltoheptaose, a difunctionalized beta-cyclodextrin. AB - 6I,6II-Diamino-6I,6II-dideoxy-cyclomaltoheptaose was prepared using the regioselective procedure described by Tabushi. The difunctionalized beta cyclodextrin crystallizes as hexadecahydrate in the orthorhombic space group P2(1)2(1)2(1), with a = 11.395(3), b = 32.989(9), c = 17.560(5) A, V = 6601 A3, Z = 4. The structure was solved by molecular replacement techniques using the program PATSEE and was refined to a conventional final R = 0.058 for the 5031 observed reflections with I > or = 3 sigma(I). The beta-CD macrocycle presents only slight differences with respect to uncomplexed hydrated or methylated beta CD. The macrocycle structure maintains an approximate seven-fold symmetry. The round shape of the cyclodextrin ring is stabilized by intramolecular O-H ... O H bonds between the secondary hydroxyl groups of neighbouring glucose residues. Along the a axis, the beta-CD molecules are arranged in columns; the macrocycles form a herring-bone pattern, so that the cavity of each beta-CD molecule is closed at each end by neighbouring molecules. The macrocycles are directly linked to each other by H-bonds involving either primary and secondary hydroxyl or amino groups of symmetry-related molecules. The resulting layers are connected to each other by a dense intermolecular hydrogen-bond network, in which solvent molecules participate. PMID- 8721736 TI - Crystal structure of anhydrous hexakis-(2,3,6-tri-O-methyl)-cyclomaltohexaose (permethyl-alpha-cyclodextrin) grown from hot water and from cold NaCl solutions. AB - An X-ray diffraction study was carried out on anhydrous permethyl-alpha cyclodextrin crystallized from aq solution at 40 degrees C, space group P2(1)2(1)2(1) with a = 15.424(9), b = 18.167(7), c = 23.128(8) A, vol = 6481(5) A3. The molecule is boat-shaped with the narrow end of the cyclodextrin cavity closed by two diametrically opposing glucose residues. These are strongly tilted towards the molecular axis, so that their primary methoxy groups form van der Waals contacts across the molecular opening. The remaining cavity volume is occupied by the methoxy group of a neighboring molecule (self-inclusion). From solutions containing 1 M NaCl, anhydrous permethyl-alpha-cyclodextrin crystallizes isomorphously at room temperature. PMID- 8721737 TI - Polymerized cyclomaltoheptaose (beta-cyclodextrin, beta-CDn) inclusion complex formation with chlorogenic acid: solvent effects on thermochemistry and enthalpy entropy compensation. AB - The inclusion of chlorogenic acid (CA) by epichlorohydrin-polymerized cyclomaltoheptaose (beta-cyclodextrin, beta-CDn) was studied with regard to temperature, solvent, and water activity aH2O approximately mole fraction = XH2O = 0.8-1 using MeOH as the diluent; 0.1 M sodium phosphate buffer). We discovered that the extreme convex curvature in K (the apparent stability constant) as a function of temperature was nearly eliminated at the lowest XH2O. The latter finding argues that this unusual CD behavior in aqueous media was due to perturbations in beta-CD's spatial organization in the polymeric matrix with temperature. Related to this we found, from the dependence of K on XH2O (K = K'XzH2O), that the beta-CDn.CA complex's stoichiometric coefficient, z, for water, varied between 5 and 8, depending on the temperature of the solution (K' = 400-800 M-1; T approximately 295-315 K). Our determinations of z were similar to those reported previously for beta-CD.(+)-limonene (z approximately 7), soluble beta-CD.CA (z approximately 6) or obtained by molecular dynamics calculations for beta-CD.CA reported herein (z approximately 5). However, beta-CDn.CA's z values did show a significant positive correlation with temperature not evident in equivalent solution experiments. Calculations of delta H and delta S at various XH2O values show linear enthalpy-entropy compensation (delta H plotted against delta S) but with a slope (Tc = theta delta H/theta delta S approximately 228 K) significantly less than Tc values determined from either standard aqueous thermodynamic experiments (Tc approximately 305 K on either beta-CD or beta-CDn) or variable XH2O (Tc approximately 272 K) experiments. To the best of our knowledge, this is the smallest Tc value detected in a multitude of CD.guest studies. This evident solvent effect on Tc strongly argues that the chemical part process of inclusion complex formation involves changes in the solvation of the beta-CDn's binding site. PMID- 8721738 TI - Synthesis and iron binding studies of myo-inositol 1,2,3-trisphosphate and (+/-) myo-inositol 1,2-bisphosphate, and iron binding studies of all myo-inositol tetrakisphosphates. AB - The first syntheses of the natural products myo-inositol 1,2,3-trisphosphate and (+/-)-myo-inositol 1,2-bisphosphate are described. The protected key intermediates 4,5,6-tri-O-benzoyl-myo-inositol and (+/-)-3,4,5,6-tetra-O-benzyl myo-inositol were phosphorylated with dibenzyl N,N-di-isopropylphosphoramidite in the presence of 1H-tetrazole and subsequent oxidation of the phosphite. The crystal structures of the synthetic intermediates (+/-)-1-O-(tert butyldiphenylsilyl)-2,3,O-cyclohexylidene-myo-inos itol and (+/-)-4,5,6-tri-O benzoyl-1-O-(tert-butyldiphenylsilyl)-2,3-O-cycl ohexylidene- myo-inositol are reported. myo-Inositol 1,2,3-trisphosphate, (+/-)-myo-inositol 1,2-bisphosphate, and all isomeric myo-inositol tetrakisphosphates were evaluated for their ability to alter HO. production in the iron-catalysed Haber-Weiss reaction. The results demonstrated that a 1,2,3-grouping of phosphates in myo-inositol was necessary for inhibition, also that (+/-)-myo-inositol 1,2-bisphosphate potentiated HO. production. myo-Inositol 1,2,3-trisphosphate resembled myo-inositol hexakisphosphate (phytic acid) in its ability to act as a siderophore by promoting iron-uptake into Pseudomonas aeruginosa. PMID- 8721739 TI - Chemical synthesis of a hexasaccharide comprising the Lewisx determinant linked beta-(1-->6) to a linear trimannosyl core and the precursor pentasaccharide lacking fucose. AB - Phenyl 2,3,4-tri-O-acetyl-6-O-chloroacetyl-1-thio-alpha,beta-mannopyranoside (5) was condensed with benzyl O-(2,3,4-tri-O-benzyl-beta-D-mannopyranosyl)-(1-->6) 2,3,4-tri-O-benzyl- alpha-D- mannopyranoside (12) in the presence of NIS-triflic acid to give, after removal of the chloroacetyl group, the key intermediate, benzyl O-(2,3,4-tri-O-acetyl-alpha-D-mannopyranosyl)-(1-->6)-O-(2,3,4-tri-O-ben zyl- beta-D-mannopyranosyl)-(1-->6)-2,3,4-tri-O-benzyl-alpha-D-mannopyranosid e (14). A similar condensation of 6 and 7 with acceptor 14, followed by the removal of protecting groups, afforded 16 and 18, respectively. These compounds are expected to be useful in specificity studies of an antibody raised against a related, synthetic antigen that we are currently investigating. PMID- 8721740 TI - Synthesis of sulfated octadecyl ribo-oligosaccharides with potent anti-AIDS virus activity by ring-opening polymerization of a 1,4-anhydroribose derivative. AB - The synthesis and anti-AIDS virus activity of sulfated octadecyl ribofuranans with medium-range molecular weights have been investigated. Selective ring opening polymerization of 1,4-anhydro-2,3-di-O-benzyl-alpha-D-ribopyranose with 10-20 mol% of boron triflouride etherate as a catalyst in a large amount of dichloromethane gave 2,3-di-O-benzyl-(1-->5)-alpha-D-ribofuranan in good yield. The molecular weight of the benzylated ribofuranan was in the range of 9 x 10(3) to 10 x 10(3). Debenzylation of the polymer followed by acetylation gave peracetylated (1-->5)-alpha-D-ribofuranans. The peracetylated ribofuranans were treated with octadecyl alcohol and a stannic chloride catalyst to afford acetylated ribofuranans having octadecyl groups at the reducing terminal. The molecular weights of the resulting acetylated octadecyl ribofuranans were below 9 x 10(3). Sulfation of the deacetylated octadecyl ribofuranans by piperidine-N sulfonic acid in dry Me2SO gave sulfated octadecyl ribofuranans with molecular weights of 3 x 10(3) to 9 x 10(3) and sulfur contents of 13.0-16.2%. The sulfated octadecyl ribofuranans had potent anti-AIDS virus activity, EC50 = 0.6-2.5 micrograms/mL (a standard curdlan sulfate showed EC50 = 0.43 micrograms/mL), and low anticoagulant activity, 4-17 units/mg (a standard dextran sulfate, 22.7 unit/mg). Structural analysis of the ribofuranans was performed by NMR at 400 and 600 MHz. PMID- 8721741 TI - The crystal structure of 6I-(6-aminohexyl)amino-6I-deoxycyclomaltoheptaose. AB - The monosubstituted cyclomaltoheptaose derivative, 6I-(6-aminohexyl)amino-6I deoxycyclomaltoheptaose, crystallizes in the orthorhombic space group P2(1)2(1)2(1) with a = 32.513(2), b = 15.3871(9), c = 15.2645(9) A, V = 7636.6(8) A3 and Z = 4. The macrocycles are spirally aligned along the twofold screw axis parallel to the c crystal axis forming polymeric-like columns. The 6-aminohexyl chain enters the cavity of an adjacent cyclomaltoheptaose moiety in the column from the secondary side and its extremity protrudes from the primary side of the latter. All the atoms of the chain exhibit high thermal motion. PMID- 8721742 TI - Structure of the O3 antigen of Stenotrophomonas (Xanthomonas or Pseudomonas) maltophilia. AB - The O antigen isolated from the lipopolysaccharide of a strain of Stenotrophomonas (Xanthomonas or Pseudomonas) maltophilia serogroup O3 was found to contain 4-acetamido-4,6-dideoxy-D-galactose, D-fucose, and N-acetyl-D glucosamine. By means of chemical degradations and NMR spectroscopy the repeating unit of the O-specific polymer was determined to be a branched trisaccharide repeating-unit of the structure shown. PMID- 8721743 TI - Elucidation of the subsite structure of bacterial saccharifying alpha-amylase and its mode of degradation of maltose. AB - The subsite structure of bacterial saccharifying alpha-amylase (BSAm) was elucidated by two methods using a series of maltooligosaccharides labeled with [14C]D-glucose at the reducing end. The rate parameter k0/Km and the cleavage frequency were obtained using the labeled substrates at sufficiently low concentrations to eliminate transglycosylation and condensation. This evaluation showed that the active center is composed of five subsites, with the catalytic site located between the 3rd and the 4th subsites from the nonreducing end. The evaluated affinity values of a subsite varied with the set of data used, which suggests some stimulation factor resulting from the chain length effect. The appearance of a time lag during the digestion of the poor substrate, maltose, was studied using radioactively labeled maltose (81.6 mM). Radioactive oligosaccharides larger than maltose were found at a significant level of more than 2% of the initial substrate in the digests, including a product peculiar to condensation, G-G*-G, as 8-10% of the maltotriose in the digests. This indicates that transglycosylation is a main side reaction (ca. 90%). A degradation pathway for maltose via maltosyl transfer was proposed, in which G3 behaves as a kind of catalyst. PMID- 8721744 TI - Rubisco surprises in dinoflagellates. PMID- 8721745 TI - To grant or not. PMID- 8721746 TI - Ectopic expression of the maize homeobox genes ZmHox1a or ZmHox1b causes pleiotropic alterations in the vegetative and floral development of transgenic tobacco. AB - The ZmHox1a and ZmHox1b (for Zea mays homeobox) genes map on chromosomes 8 and 6, respectively. Both homeobox genes encode proteins that show 91% similarity and are transcribed simultaneously in meristematic and proliferating cells of the maize plant. To gain insight into the biological function of these genes, both open reading frames were expressed in tobacco, under the control of the cauliflower mosaic virus 35S promoter. The resulting transgenic ZmHox1a or ZmHox1b plants showed identical phenotypic alterations that fall into three classes: size reduction, formation of adventitious shoots, and homeotic floral transformations. Approximately 30% of the ZmHox1-expressing plants grew to only one-third of the wild-type size, and most axillary buds gave rise to lateral shoots. Flower abnormalities included formation of petaloid stamens and development of secondary flowers within the primary gynoecium. Therefore, the ectopic expression of the maize ZmHox1 homeobox gene products affects the vegetative as well as the reproductive phase of tobacco plants. All phenotypic alterations were transmitted to the next generation. PMID- 8721747 TI - Transport of DNA into the nuclei of xenopus oocytes by a modified VirE2 protein of Agrobacterium. AB - We used Agrobacterium T-DNA nuclear transport to examine the specificity of nuclear targeting between plants and animals and the nuclear import of DNA by a specialized transport protein. Two karyophilic Agrobacterium virulence (Vir) proteins, VirD2 and VirE2, which presumably associate with the transported T-DNA and function in many plant species, were microinjected into Drosophila embryos and Xenopus oocytes. In both animal systems, VirD2 localized to the cell nuclei and VirE2 remained exclusively cytoplasmic, suggesting that VirE2 nuclear localization signals may be plant specific. Repositioning one amino acid residue within VirE2 nuclear localization signals enabled them to function in animal cells. The modified VirE2 protein bound DNA and actively transported it into the nuclei of Xenopus oocytes. These observations suggest a functional difference in nuclear import between animals and plants and show that DNA can be transported into the cell nucleus via a protein-specific pathway. PMID- 8721748 TI - Developmental and cell cycle regulation of alfalfa nucMs1, a plant homolog of the yeast Nsr1 and mammalian nucleolin. AB - We report here the isolation and characterization of the nucMs1 alfalfa cDNA, whose predicted amino acid sequence structurally resembles the yeast Nsr1 protein and animal nucleolins. These proteins consist of an N-terminal acidic domain, centrally located RNA recognition motifs (RRMs), and a C-terminal glycine- and arginine-rich domain. In comparison with animal nucleolins that contain four RRMs, NucMs1 more closely resembles the yeast Nsr1 protein, which contains only two RRMs. A NucMs1 C-terminal peptide antibody specifically recognized a 95-kD nucleolar protein in alfalfa cells that changed its localization in a cell cycle dependent manner. The nucMs1 transcript and p95nucMs1 protein levels correlated with cell proliferation, and nucMs1 gene expression was found to be induced in the G1 phase upon mitogenic stimulation of G0-arrested leaf cells. In situ hybridization analysis of different alfalfa organs during various developmental stages showed that nucMs1 gene expression is highest in root meristematic cells, but it is also found in other meristematic cells of the plant body. nucMs1 expression is tightly linked to cell proliferation but does not depend on a particular cell cycle phase. No nucMs1 expression was observed in cells that had exited the cell cycle and were undergoing differentiation or polar growth, indicating that nucMs1 may not be necessary for processes other than cell proliferation. PMID- 8721749 TI - SRK, the stigma-specific S locus receptor kinase of Brassica, is targeted to the plasma membrane in transgenic tobacco. AB - The S locus receptor kinase (SRK) gene is one of two S locus genes required for the self-incompatibility response in Brassica. We have identified the product of the SRK6 gene in B. oleracea stigmas and have shown that it has characteristics of an integral membrane protein. When expressed in transgenic tobacco, SRK6 is glycosylated and targeted to the plasma membrane. These results provide definitive biochemical evidence for the existence in plants of a plasma membrane localized transmembrane protein kinase with a known cell-cell recognition function. The timing of SRK expression in stigmas follows a time course similar to that previously described for another S locus-linked gene, the S locus glycoprotein (SLG) gene, and correlates with the ability of stigmas to mount a self-incompatibility response. Based on SRK6 promoter studies, the site of gene expression overlaps with that of SLG and exhibits predominant expression in the stigmatic papillar cells. Although reporter gene studies indicated that the SRK promoter was active in pollen, SRK protein was not detected in pollen, suggesting that SRK functions as a cell surface receptor exclusively in the papillar cells of the stigma. PMID- 8721750 TI - Leaf permease1 gene of maize is required for chloroplast development. AB - Adjacent bundle sheath and mesophyll cells cooperate for carbon fixation in the leaves of C4 plants. Mutants with compromised plastid development should reveal the degree to which this cooperation is obligatory, because one can assay whether mesophyll cells with defective bundle sheath neighbors retain C4 characteristics or revert to C3 photosynthesis. The leaf permease1-mutable1 (lpe1-m1) mutant of maize exhibits disrupted chloroplast ultrastructure, preferentially affecting bundle sheath choroplasts under lower light. Despite the disrupted ultrastructure, the metabolic cooperation of bundle sheath and mesophyll cells for C4 photosynthesis remains intact. To investigate this novel mutation, the Activator transposon-tagged allele and cDNAs corresponding to the Lpe1 mRNA from wild-type plants were cloned. The Lpe1 gene encodes a polypeptide with significant similarity to microbial pyrimidine and purine transport proteins. An analysis of revertant sectors generated by Activator excision suggests that the Lpe1 gene product is cell autonomous and can be absent up to the last cell divisions in the leaf primordium without blocking bundle sheath chloroplast development. PMID- 8721751 TI - Cold calcium signaling in Arabidopsis involves two cellular pools and a change in calcium signature after acclimation. AB - Cold shock elicits an immediate rise in cytosolic free calcium concentration ([Ca2+]cyt) in both chilling-resistant Arabidopsis and chilling-sensitive tobacco (Nicotiana plumbaginifolia). In Arabidopsis, lanthanum or EGTA caused a partial inhibition of both cold shock [Ca2+]cyt elevation and cold-dependent kin1 gene expression. This suggested that calcium influx plays a major role in the cold shock [Ca2+]cyt response and that an intracellular calcium source also might be involved. To investigate whether the vacuole (the major intracellular calcium store in plants) is involved, we targeted the calcium-dependent photoprotein aequorin to the cytosolic face of the vacuolar membrane. Cold shock calcium kinetics in this microdomain were consistent with a cold-induced vacuolar release of calcium. Treatment with neomycin or lithium, which interferes with phosphoinositide cycling, resulted in cold shock [Ca2+]cyt kinetics consistent with the involvement of inositol trisphosphate and inositide phosphate signaling in this response. We also investigated the effects of repeated and prolonged low temperature on cold shock [Ca2+]cyt. Differences were observed between the responses of Arabidopsis and N. plum-baginifolia to repeated cold stimulation. Acclimation of Arabidopsis by pretreatment with cold or hydrogen peroxide caused a modified calcium signature to subsequent cold shock. This suggests that acclimation involves modification of plant calcium signaling to provide a "cold memory." PMID- 8721752 TI - Identification of Ser-543 as the major regulatory phosphorylation site in spinach leaf nitrate reductase. AB - Spinach leaf NADH:nitrate reductase (NR) responds to light/dark signals and photosynthetic activity in part as a result of rapid regulation by reversible protein phosphorylation. We have identified the major regulatory phosphorylation site as Ser-543, which is located in the hinge 1 region connecting the cytochrome b domain with the molybdenum-pterin cofactor binding domain of NR, using recombinant NR fragments containing or lacking the phosphorylation site sequence. Studies with NR partial reactions indicated that the block in electron flow caused by phosphorylation also could be localized to the hinge 1 region. A synthetic peptide (NR6) based on the phosphorylation site sequence was phosphorylated readily by NR kinase (NRk) in vitro. NR6 kinase activity tracked the ATP-dependent inactivation of NR during several chromatographic steps and completely inhibited inactivation/phosphorylation of native NR in vitro. Two forms of NRk were resolved by using anion exchange chromatography. Studies with synthetic peptide analogs indicated that both forms of NRk had similar specificity determinants, requiring a basic residue at P-3 (i.e., three amino acids N-terminal to the phosphorylated serine) and a hydrophobic residue at P-5. Both forms are strictly calcium dependent but belong to distinct families of protein kinases because they are distinct immunochemically. PMID- 8721753 TI - Identification in vitro of a post-translational regulatory site in the hinge 1 region of Arabidopsis nitrate reductase. AB - Nitrate reductase (NR) is rapidly inactivated by phosphorylation of serine residues in response to loss of light or reduction in CO2 levels. To identify sites within NR protein that play a role in this post-translational regulation, a heterologous expression system and an in vitro inactivation assay for Arabidopsis NR were developed. Protein extracts containing NR kinases and inhibitor proteins were prepared from an NR-defective mutant that had lesions in both the NIA1 and NIA2 NR genes of Arabidopsis. Active NR protein was produced in a Pichia pastoris expression system. Incubation of these two preparations resulted in a Mg-ATP dependent inactivation of NR that was reversed with EDTA. Mutant forms of NR were constructed, produced in P. pastoris, and tested in the in vitro inactivation assay. Six conserved serine residues in the hinge 1 region of NR, which separates the molybdenum cofactor and heme domains, were specifically targeted for mutagenesis because they are located in a potential regulatory region identified as a target for NR kinases in spinach. A change in Ser-534 to aspartate was found to block NR inactivation; changes in the other five serines had no effect. The aspartate that replaced Ser-534 did not appear to mimic a phosphorylated serine but simply prevented the NR from being inactivated. These results identify Ser 534, located in the hinge 1 of NR and conserved among higher plants NRs, as an essential site for post-translational regulation in vitro. PMID- 8721754 TI - The SAL1 gene of Arabidopsis, encoding an enzyme with 3'(2'),5'-bisphosphate nucleotidase and inositol polyphosphate 1-phosphatase activities, increases salt tolerance in yeast. AB - A cDNA library in a yeast expression vector was prepared from roots of Arabidopsis exposed to salt and was used to select Li(+)-tolerant yeast transformants. The cDNA SAL1 isolated from one of these transformants encodes a polypeptide of 353 amino acid residues. This protein is homologous to the HAL2 and CysQ phosphatases of yeast and Escherichia coli, respectively. Partial cDNA sequences in the data bases indicate that rice produces a phosphatase highly homologous to SAL1 and that a second gene homologous to SAL1 exists in Arabidopsis. The SAL1 protein expressed in E. coli showed 3'(2'),5'-bisphosphate nucleotidase and inositol polyphosphate 1-phosphatase activities. In yeast, SAL1 restored the ability of a hal2/met22 mutant to grow on sulfate as a sole sulfur source, increased the intracellular Li+ tolerance, and modified Na+ and Li+ effluxes. We propose that the product of SAL1 participates in the sulfur assimilation pathway as well as in the phosphoinositide signaling pathway and that changes in the latter may affect Na+ and Li+ fluxes. PMID- 8721756 TI - Receptors and medicinal chemistry: new methods and perspectives. PMID- 8721755 TI - Rubisco in marine symbiotic dinoflagellates: form II enzymes in eukaryotic oxygenic phototrophs encoded by a nuclear multigene family. AB - Genes encoding ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) were cloned from dinoflagellate symbionts (Symbiodinium spp) of the giant clam Tridacna gigas and characterized. Strikingly, Symbiodinium Rubisco is completely different from other eukaryotic (form I) Rubiscos: it is a form II enzyme that is approximately 65% identical to Rubisco from Rhodospirillum rubrum (Rubisco forms I and II are approximately 25 to 30% identical); it is nuclear encoded by a multigene family; and the predominantly expressed Rubisco is encoded as a precursor polyprotein. One clone appears to contain a predominantly expressed Rubisco locus (rbcA), as determined by RNA gel blot analysis of Symbiodinium RNA and sequencing of purified Rubisco protein. Another contains an enigmatic locus (rbcG) that exhibits an unprecedented pattern of amino acid replacement but does not appear to be a pseudogene. The expression of rbcG has not been analyzed; it was detected only in the minor of two taxa of Symbiodinium that occur together in T. gigas. This study confirms and describes a previously unrecognized branch of Rubisco's evolution: a eukaryotic form II enzyme that participates in oxygenic photosynthesis and is encoded by a diverse, nuclear multigene family. PMID- 8721757 TI - Synthesis and pharmacological study of some new beta-(dialkylaminomethyl)- gamma butyrolactones and their tetrahydrofuran analogues. AB - This paper describes the synthesis of beta-(dialkylaminomethyl)-gamma- butyrolactones (6 and 15) and their tetrahydrofuran analogs 7 and 16. Their convulsant activity was studied on mice and could display an antiGABAergic component, but, unlike the alpha-(dialkylaminomethyl)- gamma-butyrolactones, no antiglycinergic component was detected. The possibility of an activation of the glutamatergic receptors (NMDA), by indirect stimulation of their glycinergic site, by the tetrahydrofurans analogs 7 could be considered. These compounds exhibited, at low doses (1/3 to 1/20 of their convulsant doses), an anticonvulsant action in the maximal electroshock test and this is in agreement with the abovementioned possibility. PMID- 8721758 TI - Structure-activity relationships in 1,4-benzodioxan-related compounds. 5. Effects of modification of the side chain on alpha-adrenoreceptor blocking activity. AB - The observation that the insertion of a phenyl ring at position 3 of WB 4101 (1) afforded a potent and selective alpha 1-adrenoreceptor antagonist, phendioxan (2), prompted us to investigate the effect on alpha-adrenoreceptor blocking activity of replacing a hydrogen atom at the 2-position or on the 2-ylmethyl moiety of 1 and 2 by a methyl, ethyl or phenyl group. Thus compounds 3-11 were synthesized and their blocking activity and relative selectivity on alpha 1- and alpha 2-adrenoreceptors were evaluated in the isolated rat vas deferens in comparison to 1 and 2. The results of such investigation showed that this structural modification caused a decrease in affinity for alpha-adrenoreceptors. PMID- 8721759 TI - (E)-(methyloxyimino)acetamides as analogues of neuroleptic benzamides: synthesis and D2-dopaminergic binding affinity. AB - Some type C (E)-(methyloxyimino)acetamides were synthesised as analogues of type A neuroleptic and antipsychotic benzamides, in which the aromatic group is substituted by a methyloxyiminomethyl moiety with the E configuration (CH2ON = CH, E-MOIMM). Type C compounds were tested for their D2-dopaminergic binding affinity in order to obtain an indication of their potential neuroleptic and antipsychotic properties. Biological results showed that only a few aryl substituted E-MOIM derivatives possess a certain affinity for the D2-dopaminergic receptor, at least one order of magnitude lower than that of metoclopramide and sulpiride. PMID- 8721760 TI - Synthesis and biological evaluation of mono- and bis-[(alkylamino)alkylamino] substituted thienopyridopyridazines, a new class of potential antitumor agents. AB - A new class of potential antitumor agents, provided with thieno[2',3':5,6]pyrido[2,3-d]pyridazin-9(4H)-one nucleus as chromophore, was synthesized. Thus, the suitable amines were reacted, in different conditions, with 5,8-dichlorothieno[2',3':5,6]pyrido[2,3- d]pyridazin-9(4H)-one (5) to afford the 8-alkylamino derivatives 6a-f, the 5-alkylamino derivatives 7a-f, and the 5,8 bis-alkylamino derivatives 8a,b. Selected compounds were evaluated for cytotoxic potency in vitro against the human colon adenocarcinoma HT 29 cell line and studied in DNA binding assays. The cytotoxic potency versus HT29 was also correlated with binding affinity for calf thymus DNA. PMID- 8721761 TI - 3-Diazopyrroles. Part 6. Mutagenic activity of 3-diazopyrroles in Streptomyces coelicolor A3(2) during various phases of growth. AB - 3-Diazopyrroles, a class of compounds particularly interesting from a chemical and biological point of view, were assayed for their ability to induce gene mutations employing back mutation (his+ reversion) test in the philamentous bacterium Streptomyces coelicolor at various time during life cycle. Our results suggest that in evaluating the mutagenicity and toxicity of chemicals in Streptomyces system it is important to consider factors such as growth phase. Furthermore in this series of diazopyrroles a relationship between toxicity, mutagenicity and chemical structure was found. The observed mutagenic activity can be the molecular basis for the appearance of antitumor activity. PMID- 8721762 TI - Synthesis of 1,2-disubstituted benzimidazole-5(6)-carboxamides and evaluation of their antimicrobial activity. AB - A series of 14, N'-(N,N-dialkylaminoethyl)-benzimidazole-5(6) or 5-carboxyamides (1-14), having several substituents on the azole and benzene nuclei, were prepared and evaluated in vitro for antimicrobial activity. The precursor benzimidazolecarboxylic acids (15-27) were prepared via oxidative condensation of diaminobenzoic acids and several aldehydes with cupric ion. Compounds 11-14 were prepared by selective regioisomer synthesis. All carboxamides were prepared from the corresponding acids and N,N-dialkylethylenediamine. Antibacterial and antifungal activities were determined as MIC values. Of the synthesized compounds 1-10, 6 and 10 were found to be the most favourable. In order to clarify the effect of the substituents at N1 on antimicrobial activity, 12 was prepared by p chlorobenzyl substitution of compound 6, and increased activity was shown. Compounds 13 and 14, which were prepared by replacement with more bulky alkyl groups on the tert-N atom than 12, gave the best results. PMID- 8721763 TI - Synthesis of some novel pyrazoline and cyanopyridine derivatives as antimicrobial agents. AB - Substituted chalcones 2a-j prepared by the treatment of 4'(p chlorophenylsulphonamido)acetophenone 1 with araldehydes, on condensation with hydrazine hydrate in ethanol and acetic acid provided the desired pyrazolines of the type 3a-j and 4a-j. The same chalcones 2a-j made to react with malononitrile in the presence of ammonium acetate yielded cyanopyridines 5a-j. The structure was confirmed by elemental analyses, IR, NMR and MS spectral studies. The pharmacological profile of the compounds synthesised is described. Most of the compounds displayed their maximum activity against E. coli. Compounds 2b, 2d, 5b, 5d, 5e, 5h and 5i have been selected for their agrochemical screening by DuPont Agricultural Products, U.S.A. PMID- 8721764 TI - Synthesis and antimycobacterial activity of some indole derivatives of pyridine-2 carboxamidrazone and quinoline-2-carboxamidrazone. AB - A series of pyridine-2-carboxamidrazone and quinoline-2-carboxamidrazone derivatives containing the indole moiety was prepared. Some of the synthesized compounds showed an interesting in vitro antimycobacterial activity against a strain of Mycobacterium tuberculosis. PMID- 8721765 TI - Synthesis and antimicrobial activity of some 2,5-disubstituted 1,3,4,-thiadiazole derivatives. AB - A series of 5-substituted 2-arylamino-1,3,4-thiadiazole derivatives was prepared. The antimicrobial activity of these compounds against some strains of bacteria and a strain of Candida albicans was determined, together with that of the corresponding thiosemicarbazone derivatives, which are intermediates in the synthetical procedure. PMID- 8721766 TI - Synthesis and antimicrobial activity of benzo[a]dihydrocarbazole and benzotetrahydrocyclohept[1,2-b]indole derivatives. AB - A certain number of benzodihydrocarbazoles and benzotetrahydrocycloheptindoles were synthesized and tested for their antimicrobial activity. Compounds substituted on the nitrogen of the tetracyclic systems with a dimethylaminopropyl chain showed an antibacterial activity directed almost exclusively towards Gram positive bacteria, while their N-unsubstituted analogs were completely inactive. PMID- 8721767 TI - Fungicidal activity of some o-nitrophenyl-hydrazones. AB - The antimycotic activity of 16 o-nitrophenylhydrazones against strains of Hansenula anomala, Saccharomyces cerevisiae, Candida parapsylosis, and Cryptococcus albidus was tested. All 16 compounds inhibited growth of the yeast strains. The inhibitory activity of the 4 methyl-derivatives substituted on the aromatic nucleus was particularly significant. PMID- 8721768 TI - Acceleration of experimental diabetic retinopathy in the rat by omega-3 fatty acids. AB - Omega-3 fatty acids exert several important biological effects on factors that may predispose to diabetic retinopathy. Potential pathogenetic mechanisms include platelet dysfunction, altered eicosanoid production, increased blood viscosity in association with impaired cell deformability and pathologic leucocyte/endothelium interaction. Therefore, we tested whether a 6-month administration of fish oil (750 mg Maxepa, 5 times per week), containing 14% eicosapentaenoic acid (EPA) and 10% docosahexaenic acid, could inhibit the development of experimental retinopathy of the streptozotocin-diabetic rat. The efficiency of fish oil supplementation was evaluated by measuring EPA concentrations in total, plasma and membrane fatty acids and by measuring the generation of lipid mediators (leukotrienes and thromboxanes). Retinal digest preparations were quantitatively analysed for pericyte loss, and the formation of acellular capillaries. Omega-3 fatty acid administration to diabetic rats resulted in a twofold increase of EPA 20:5 in total fatty acids, and a reduction of the thromboxane ratio from 600 (untreated diabetic rats) to 50 (treated diabetic rats). Despite these biochemical changes, diabetes-associated pericyte loss remained unaffected and the formation of acellular, occluded capillaries was increased by 75% in the fish oil treated diabetic group (115.1 +/- 26.8; untreated diabetic 65.2 +/- 15.0 acellular capillary segments/mm2 of retinal area). We conclude from this study that dietary fish oil supplementation may be harmful for the diabetic microvasculature in the retina. PMID- 8721769 TI - Islet-cell regeneration in the diabetic hamster pancreas with restoration of normoglycaemia can be induced by a local growth factor(s). AB - Partial pancreatic duct obstruction in the hamster leads to the induction of endocrine-cell differentiation and new islet formation. We prepared cytosolic extracts from the partially obstructed pancreas and identified one, which when administered i.p., produced significant increases in the incorporation of tritiated thymidine by ductular and islet cells, as well as a corresponding increase in islet mass. In this study, we evaluate the ability of this extract to reverse streptozotocin diabetes mellitus. Hamsters were treated i.p. twice daily for 7 weeks with either 0.9% NaCl (saline) (n = 10) or a cytosol extract (n = 10) prepared previously from partially obstructed hamster pancreata. All animals in the cytosol group survived vs only 60% of the saline group (p = 0.02). Random blood glucose levels were greater than 22.2 mmol/l in 90% of the saline group vs 40% in the cytosol group (p < 0.05). Pancreatic tissue from the surviving saline animals and from persistently hyperglycaemic cytosol-treated animals, showed intra-cytoplasmic vacuolation of islet cells, a characteristic lesion of sustained hyperglycaemic states. Vacuolation was not observed in normoglycaemic extract treated animals. Islets in hyperglycaemic animals demonstrated a profound decrease or absence of immunoreactive insulin, compared to an abundance of immunoreactive beta cells in cytosol-treated animals that reverted to normoglycaemia. In this group, single cells or nests of cells stained for insulin on glucagon cells were identified in ductal epithelium in association with cells budding from the duct. Morphometric analysis of pancreata in reverted cytosol treated animals showed a new population of small islets compared with saline controls and an increased islet mass. In summary, streptozotocin diabetes can be reversed by new islet formation induced by local pancreatic growth factors, the exact nature of which remains to be determined. PMID- 8721770 TI - Inhibition of development of peripheral neuropathy in streptozotocin-induced diabetic rats with N-acetylcysteine. AB - N-acetylcysteine (NAC) is a precursor of glutathione (GSH) synthesis, a free radical scavenger and an inhibitor of tumour necrosis factor alpha (TNF). Because these functions might be beneficial in diabetic complications, in this study we examined whether NAC inhibits peripheral neuropathy. Motor nerve conduction velocity (MNCV) was significantly decreased in streptozotocin-induced-diabetic Wistar rats compared to control rats. Oral administration of NAC reduced the decline of MNCV in diabetic rats. Structural analysis of the sural nerve disclosed significant reduction of fibres undergoing myelin wrinkling and inhibition of myelinated fibre atrophy in NAC-treated diabetic rats. NAC treatment had no effect on blood glucose levels or on the nerve glucose, sorbitol and cAMP contents, whereas it corrected the decreased GSH levels in erythrocytes, the increased lipid peroxide levels in plasma and the increased lipopolysaccharide-induced TNF activity in sera of diabetic rats. Thus, NAC inhibited the development of functional and structural abnormalities of the peripheral nerve in streptozotocin-induced diabetic rats. PMID- 8721771 TI - Prevention of decline in renal function in the diabetic db/db mouse. AB - We recently reported that when diabetic db/db mice, which develop glomerular pathology resembling that in human diabetes mellitus, are treated with monoclonal antibodies (A717) that neutralize the effects of excess glycated albumin, there is an amelioration of mesangial expansion, renal overexpression of mRNAs encoding for the extracellular matrix proteins collagen IV and fibronectin and proteinuria. These findings suggested that A717 might also retard the development of compromised renal function in this animal model. To examine this possibility, serum creatinine and blood urea nitrogen (BUN) were measured in diabetic db/db mice and their non-diabetic db/m littermates before and after an 8-week course of treatment with A717 or irrelevant murine immunoglobulin (MIg). Early in the course of diabetes, BUN and serum creatinine concentrations did not significantly differ from those in the db/m littermates, but were significantly increased after 10 weeks of sustained hyperglycaemia. Treatment of db/db mice with A717 prevented the rise in creatinine and attenuated the elevation in BUN. A717 also prevented the decrease in creatinine clearance observed in diabetic compared with non diabetic animals (2.2 +/- 0.8 vs 4.1 +/- 0.3 vs 5.0 +/- 1.1 ml/h in db/db vs db/db-A717 vs db/m, respectively). MIg did not alter the change in renal function with time in db/db mice. Taken together with our previous results, the present findings indicate that the diabetic db/db mouse develops changes in renal function and structure that parallel the course of human diabetic nephropathy in nature and chronology and demonstrate, for the first time, that therapy directed against increased glycated albumin can prevent the decline in renal function in this rodent model of genetic diabetes. PMID- 8721773 TI - Soluble, fatty acid acylated insulins bind to albumin and show protracted action in pigs. AB - We have synthesized insulins acylated by fatty acids in the epsilon-amino group of LysB29. Soluble preparations can be made in the usual concentration of 600 nmol/ml (100 IU/ml) at neutral pH. The time for 50% disappearance after subcutaneous injection of the corresponding TyrA14(125I)-labelled insulins in pigs correlated with the affinity for binding to albumin (r = 0.97), suggesting that the mechanism of prolonged disappearance is binding to albumin in subcutis. Most protracted was LysB29-tetradecanoyl des-(B30) insulin. The time for 50% disappearance was 14.3 +/- 2.2 h, significantly longer than that of Neutral Protamine Hagedorn (NPH) insulin, 10.5 +/- 4.3 h (p < 0.001), and with less inter pig variation (p < 0.001). Intravenous bolus injections of LysB29-tetradecanoyl des-(B30) human insulin showed a protracted blood glucose lowering effect compared to that of human insulin. The relative affinity of LysB29-tetradecanoyl des-(B30) insulin to the insulin receptor is 46%. In a 24-h glucose clamp study in pigs the total glucose consumptions for LysB29-tetradecanoyl des-(B30) insulin and NPH were not significantly different (p = 0.88), whereas the times when 50% of the total glucose had been infused were significantly different, 7.9 +/- 1.0 h and 6.2 +/- 1.3 h, respectively (p < 0.04). The glucose disposal curve caused by LysB29-tetradecanoyl des-(B30) insulin was more steady than that caused by NPH, without the pronounced peak at 3 h. Unlike the crystalline insulins, the soluble LysB29-tetradecanoyl des-(B30) insulin does not elicit invasion of macrophages at the site of injection. Thus, LysB29-tetradecanoyl des-(B30) insulin might be suitable for providing basal insulin in the treatment of diabetes mellitus. PMID- 8721772 TI - Angiotensinase A gene expression and enzyme activity in isolated glomeruli of diabetic rats. AB - One of the characteristics of early diabetic nephropathy is glomerular hyperfiltration and hyperperfusion. Many factors have been suggested to induce glomerular hyperperfusion among which are an increased production of vasodilatory prostanoids, an increased synthesis of nitric oxide, a reduced responsiveness of afferent glomerular arterioles to vasoconstrictor stimuli due to diabetic metabolic disturbances and a decreased receptor density for angiotensin II. It has been known for years that angiotensin II is formed locally due to the local activation of the renin angiotensin system. The local angiotensin II concentration, however, is not only regulated by the synthesis rate but also by the local degradation through activation of an aminopeptidase. The main finding of the present study was that the mRNA expression and activity of the angiotensin II degrading enzyme, angiotensinase A, was increased twofold in diabetic rats at 5 weeks and that the increase in mRNA expression was suppressed by insulin therapy and short-term treatment with the angiotensin II antagonist saralasin, whereas angiotensinase A enzyme activity was only reduced by saralasin and not by insulin. These results demonstrate that the angiotensin II degrading exopeptidase angiotensinase A is activated in diabetic glomeruli. This increased activity may be an additional mechanism to explain glomerular hyperfiltration and hyperperfusion in early diabetic nephropathy. PMID- 8721775 TI - Intra-individual variation of glucose, specific insulin and proinsulin concentrations measured by two oral glucose tolerance tests in a general Caucasian population: the Hoorn Study. AB - We studied the intra-individual variation in plasma glucose, specific serum insulin and serum proinsulin concentrations, measured by two 75-g oral glucose tolerance tests in an age, sex, and glucose tolerance stratified random sample from a 50-74-year-old Caucasian population without a history of diabetes mellitus. The intra-individual variation was assessed by the standard deviation of the test-retest differences (SDdif). For subjects with normal (n = 246), impaired glucose tolerance (n = 198), and newly detected diabetes (n = 80) classified at the first test, the following (SDdif/median level of individual average scores) were found: fasting glucose: 0.4/5.4, 0.5/5.9 and 0.7/7.2 mmol/l; 2-h glucose: 1.3/5.6, 1.8/8.5 and 2.3/12.8 mmol/l; fasting insulin: 23/76, 32/89 and 30/116 pmol/l; 2-h insulin: 190/303, 278/553 and 304/626 pmol/l; fasting proinsulin: 4/8, 6/13 and 9/18 pmol/l; 2-h proinsulin: 19/49, 23/84 and 33/90 pmol/l, respectively. In both glucose, proinsulin and insulin concentrations the total intra-individual variation was predominantly determined by biological variation, whereas analytical variation made only a minor contribution. The SDdif can easily be interpreted, as 95% of the random test-retest differences will be less than 2.SDdif, or in terms of percentage, less than (2.SDdif/median level of individual average score) 100. Therefore, for subjects with normal glucose tolerance, 95% of the random test-retest differences will be less than 15% (fasting glucose), 46% (2-h glucose), 61% (fasting insulin), 125% (2-h insulin), 100% (fasting proinsulin) and 78% (2-h proinsulin) of the median value of the individual average scores. No substantial independent association of either age, gender or obesity with the intra-individual variation in glucose, proinsulin, or insulin concentrations was found. PMID- 8721774 TI - Delayed intracellular dissociation of the insulin-receptor complex impairs receptor recycling and insulin processing in cultured Epstein-Barr virus transformed lymphocytes from insulin-resistant subjects. AB - Insulin-receptor internalization and processing are defective in insulin resistant subjects. To assess the reversibility of these defects, we cultured Epstein-Barr virus-transformed-lymphoblasts from six normal, six obese, and six non-insulin-dependent diabetic (NIDDM) subjects in media containing low (5 mmol/l) or high (25 mmol/l) glucose concentrations, and studied the insulin receptor internalization and processing in vitro. In cells from normal, obese, and NIDDM subjects cultured in low glucose concentrations, exposure to 100 nmol/l insulin for 30 min at 37 degrees C reduced cell-surface 125I-insulin binding to a similar extent (82 +/- 2, 77 +/- 5, and 82 +/- 5% of initial values, respectively). The same results were obtained with cells cultured in high glucose concentrations. In cells cultured under both glucose conditions, and exposed to 100 nmol/l insulin for 30 min at 37 degrees C, a complete recovery of the initial 125I-insulin binding was observed in normal but not in obese and NIDDM subjects. Release of intracellular insulin and its degradation in vitro was determined by incubating cells with 600 pmol/l of 125I-insulin for 60 min at 37 degrees C, acid washing cells, and re-incubating in insulin-free buffer at 37 degrees C. The radioactivity released by cells was characterized by trichloroacetic acid precipitability, Sephadex G-50 column chromatography, and re-binding to fresh cells. Rates of release of internalized radioactivity were reduced in obese and NIDDM subjects (t1/2 = 61 +/- 9 min, p < 0.02; 58 +/- 10 min, p < 0.05; and 38 +/ 4 min in obese, NIDDM, and normal subjects, respectively). The percentage of intact insulin released from cells was significantly higher in obese and NIDDM subjects than in the normal subjects. The t1/2 of intracellular dissociation of insulin-receptor complexes measured by a polyethylene glycol assay was lower in normal (6 +/- 1 min) than in obese (12 +/- 2 min, p < 0.03) and NIDDM subjects (14 +/- 3 min, p < 0.02). The results suggest that in insulin-resistant subjects a primary defect in intracellular dissociation of insulin is responsible for alterations of receptor recycling and insulin processing. PMID- 8721776 TI - C-peptide stimulates glucose transport in isolated human skeletal muscle independent of insulin receptor and tyrosine kinase activation. AB - We have previously demonstrated that C-peptide stimulates glucose transport in skeletal muscle from non-diabetic subjects in a dose-dependent manner. To further elucidate the mechanism by which C-peptide activates glucose transport, we investigated the influence of human recombinant C-peptide on receptor and post receptor events involved in the glucose transport process. Human skeletal muscle specimens were obtained from the vastus lateralis by means of an open biopsy procedure. Stimulation of isolated muscle strips from healthy control subjects with supra-physiological concentrations of insulin (6,000 pmol/l) and C-peptide (2,500 pmol/l), did not further augment the twofold increase in the rate of 3-o methylglucose transport induced by either stimulus alone. C-peptide did not displace 125I-insulin binding from partially purified receptors, nor did it activate receptor tyrosine kinase activity. Tyrosine-labelled 125I-C-peptide did not bind specifically to crude membranes prepared from skeletal muscle, or to any serum protein other than albumin. The beta-adrenergic receptor stimulation with isoproterenol inhibited insulin- but not C-peptide-mediated 3-o-methylglucose transport by 63 +/- 18% (p < 0.01), whereas the cyclic AMP analogue, Bt2cAMP, abolished the insulin- and C-peptide-stimulated 3-o-methylglucose transport. C peptide (600 pmol/l) increased 3-o-methylglucose transport 1.8 +/- 0.2-fold in skeletal muscle specimens from patients with insulin-dependent diabetes mellitus. In conclusion, C-peptide stimulates glucose transport by a mechanism independent of insulin receptor and tyrosine kinase activation. In contrast to the effect on insulin-stimulated glucose transport, catecholamines do not appear to have a counter regulatory action on C-peptide-mediated glucose transport. PMID- 8721777 TI - Association of the glycogen synthase locus on 19q13 with NIDDM in Pima Indians. AB - Skeletal muscle glycogen synthase (encoded by GYS1 on chromosome 19q13.3) is the rate-limiting enzyme in insulin-mediated non-oxidative glucose disposal. Our previous studies have demonstrated an impairment of insulin-stimulated GYS1 activities in insulin-resistant Pima Indians, and associations of non-insulin dependent diabetes mellitus (NIDDM) with the GYS1 locus were reported recently in Finnish and Japanese populations. We have performed linkage and association analyses of GYS1 and seven additional DNA markers on 19q with NIDDM, and with parameters of insulin action in the Pima Indians. We have found a significant association of NIDDM with GYS1 genotypes (p = 0.009), and with common GYS1 alleles (p = 0.022) in the Pima Indians. We have performed a detailed comparative analysis of the GYS1 gene, mRNA, and protein product in insulin-sensitive and insulin-resistant Pima Indians. No mutations in GYS1 coding sequences were detected; nor did we find alterations of GYS1 mRNA expression or of its basal enzymatic activity in insulin-resistant Pima Indians. These results contrasted with a 25% reduction of immunoreactive protein in insulin-resistant subjects as detected by Western blotting with an antibody specific for the C-terminal end of GYS1 (t-test p = 0.024; Wilcoxon's rank-sum test, p = 0.04). Because no mutations were detected in the DNA encoding this epitope, the difference in immunoreactivity may reflect post-translational modification(s) of the protein rather than a difference in the gene itself, or it could have occurred by chance. We conclude that our data do not indicate alterations in the GYS1 gene as the cause for the observed association, and that a different locus near GYS1 may be the contributing genetic element. PMID- 8721778 TI - Variant sequences of the Hexokinase II gene in familial NIDDM. AB - Hexokinase II (HKII) plays a central role in the intracellular metabolism of glucose in skeletal muscle, catalysing the phosphorylation of glucose to glucose 6-phosphate. It is therefore considered to be a potentially important candidate gene in the development of insulin resistance and non-insulin-dependent diabetes mellitus (NIDDM). The aim of this study was to screen the HKII gene for mutations in NIDDM subjects from insulin-resistant families. Insulin sensitivity was assessed in unaffected first degree relatives from families with two or more living NIDDM subjects, and 15 families were identified as being insulin resistant. In 15 NIDDM subjects (one from each family) and 4 normoglycaemic control subjects, all 18 exons of the HKII gene were amplified by the polymerase chain reaction, and the products screened for mutations using a combination of single-stranded conformational polymorphism analysis and direct sequencing. Six sequence variations were detected in the NIDDM subjects; four silent polymorphisms [GAT vs GAC at codon 251 in exon 7, AAT vs AAC at codon 692 in exon 15, CCG vs CCC at codon 736 in exon 15, and CTG vs CTA at codon 766 in exon 16]; a single base change [T-->C], 22 base pairs distal to the exon-intron junction of exon 17 in the 5'-splice donor; and a single amino acid substitution [Gln142- >His] in exon 4, which was identified in 6 of the 15 NIDDM subjects. The frequency of the mutated codon 142 allele however, was comparable between NIDDM subjects with familial NIDDM (n = 56) and normoglycaemic control subjects (n = 48) (18.8% and 14.6% for NIDDM subjects and control subjects respectively; chi 2 = 0.6, p > 0.25). In addition, measures of insulin sensitivity were comparable in normal glucose tolerant subjects with (n = 20) and without (n = 40) the codon 142 polymorphism. IN CONCLUSION: (1) mutations in the coding regions of the HKII gene are unlikely to be major determinants in the development of insulin resistance and familial NIDDM; although (2) the influence of the codon 142 mutation in combination with other abnormalities of the insulin-signalling pathway on insulin action remain to be addressed. PMID- 8721779 TI - Arterio-venous shunting and proliferating new vessels in acute painful neuropathy of rapid glycaemic control (insulin neuritis). AB - Insulin neuritis, or painful neuropathy following rapid improvement in glycaemic control, is well recognised but its aetiology is unclear. An understanding of the processes involved in the genesis of acute painful neuropathy of rapid glycaemic control may give an insight into the early pathogenetic factors leading to diabetic nerve damage in general. We have identified five subjects with insulin neuritis including one who developed severe autonomic neuropathy following treatment with insulin. Subjects underwent: 1) assessment of neuropathic symptom and deficit scores; 2) quantitative sensory and electrophysiological studies and 3) sural nerve epineurial vessel photography and fluorescein angiography in vivo. The sural nerve photographs were independently graded by an ophthalmologist. All subjects with insulin neuritis presented with severe sensory symptoms but clinical examination and electrophysiological tests were normal except in the subject with the severe autonomic neuropathy in whom all the tests were abnormal. On nerve photography, there was an abundance of epineurial nutrient vessels although these showed severe abnormalities including arteriolar attenuation, tortuosity and arterio-venous shunting in all subjects. Proliferating neural 'new vessels' which bear striking similarities to those found in the retina and that were more leaky to fluorescein than normal vessels, were observed in three subjects. Venous distension and/or tortuosity was also observed in three subjects and this was most marked in the subject with severe autonomic neuropathy. This study shows that epineurial nutrient vessel anatomy is abnormal in subjects with acute painful neuropathy of rapid glycaemic control, a condition previously thought to be purely metabolic in origin. The presence of epineurial arterio venous shunting and a fine network of vessels resembling the new vessels of the retina, may lead to a 'steal' effect rendering the endoneurium ischaemic. This process may be important in the genesis of neuropathic pain, and further supports the importance of vascular factors in the pathogenesis of diabetic neuropathy. PMID- 8721780 TI - Elevated von Willebrand factor antigen predicts deterioration in diabetic peripheral nerve function. AB - We have studied the temporal relationship of plasma von Willebrand Factor (vWF), a marker of endothelial damage, with the development of complications in 63 young diabetic patients (56 of whom were insulin-dependent) who took part in a prospective study. Results are presented from baseline to follow-up. In the group as a whole, no significant changes were found in any autonomic function tests, temperature discrimination threshold or nerve conduction velocities. Median motor and peroneal latency were significantly increased, while median motor, peroneal motor and sural sensory potentials were significantly decreased at follow-up compared with baseline (p < 0.001). There was a significant fall in mean plasma vWF antigen and ristocetin co-factor activity in the entire group. Within the group, we identified eight patients whose peroneal motor and sural sensory conduction velocities had decreased by over 2 ms-1. In these patients (Group A), baseline vWF antigen and activity were significantly higher than in the rest of the patients (Group B), (p = 0.04) and vWF antigen was still significantly higher after 3 years (p = 0.02). There were no differences between the groups in incidence of retinopathy, urinary albumin excretion rate or macrovascular disease. To assess the influence of glycaemic control on vWF, we first compared a matched group (C) of diabetic patients with similar HbA1 to that of group A, but with normal nerve conduction velocities: vWF was still significantly higher in group A compared with group C (p = 0.02). Furthermore, hierarchical regression showed that vWF predicted deteriorating nerve function independently of glycaemic control or the type of diabetes. Endothelial dysfunction may be associated with development of peripheral neuropathy in young diabetic patients. PMID- 8721781 TI - The impact of reversal of hypoxia by revascularization on the peripheral nerve function of diabetic patients. AB - Hypoxia is considered to be one of the main aetiopathogenic factors of diabetic neuropathy. We have examined the effects of the reversal of hypoxia, achieved by revascularization, on peripheral nerve function in diabetic patients with or without clinical neuropathy. Fifty-six patients [mean age 62 (range 30-74) years, 44 (79%) males, 15 (27%) with insulin-dependent diabetes of 20 years (range 1-57) duration, and creatinine level 92.8 +/- 30.9 mumol/l (mean +/- SD)] were tested pre-operatively while 30 (54%) were reexamined at least 6 weeks post-operatively. At baseline the leg scheduled for operation showed worse measurements compared to the control leg when tested for Semmes-Weinstein monofilaments, peroneal motor conduction velocity (PMCV) (33.7 +/- 7.18 vs 35.7 +/- 6.09 m.s-1, p < 0.05) and transcutaneous oxygen tension (37.4 +/- 24.6 vs 52.0 +/- 21.5 mm Hg, p < 0.0001) while no differences were found in the vibration perception threshold and leg temperature. When baseline and post-operative measurements were later compared in the operated leg, no differences were noticed in the vibration perception threshold, PMCV and Semmes-Weinstein monofilaments but the transcutaneous oxygen tension increased significantly (32.7 +/- 27.1 vs 64.6 +/- 14.5 mm Hg, p < 0.001). No differences were noticed in any of the above parameters in the contralateral leg. No correlations were found between changes in transcutaneous oxygen tension and PMCV values measured at baseline and at the follow-up visit in either leg. Similar results were found when patients were stratified according to severity of neuropathy, ischaemia and the level of the bypass. We conclude that although there is greater impairment of nerve function in the more ischaemic leg, reversal of hypoxia does not result in any significant improvement of the nerve function measurements. PMID- 8721782 TI - Association of Trp64Arg mutation of the beta3-adrenergic-receptor with NIDDM and body weight gain. AB - A possible pathogenic mutation in the beta 3-adrenergic-receptor gene (Trp64Arg) has been reported to be associated with an earlier age of onset of non-insulin dependent diabetes mellitus (NIDDM) and clinical features of the insulin resistance syndrome in Pima Indian, Finnish and French subjects. Since marked heterogeneity has been reported in the association of mutations of candidate genes with NIDDM between Japanese and other ethnic groups, we investigated the association of Trp64Arg with NIDDM in Japanese subjects. The allele frequency of the mutation (Arg) was slightly, but not significantly, higher in NIDDM than in control subjects (70 out of 342 alleles [20.5%] vs 40 out of 248 [16.1%], respectively, p > 0.2). When our data were combined with those of Pima Indian and Finnish subjects, however, the Arg/Arg genotype was significantly associated with NIDDM as compared with the other two genotypes (p < 0.005, relative risk [RR] 2.13, 95% confidence interval [CI] 1.28-3.55). The Arg allele was also associated with NIDDM (p < 0.05, RR 1.27, 95% CI 1.06-1.52). Japanese subjects homozygous for the mutation had a significantly higher body mass index (mean +/- SD: 25.5 +/ 3.9 kg/m2) than heterozygotes (22.6 +/- 4.1, p < 0.05) and normal homozygotes (22.8 +/- 3.8, p < 0.05). NIDDM patients homozygous for the mutation tended to have an earlier age of onset of NIDDM than those with other genotypes. These data suggest that the Trp64Arg mutation not only contributes to weight gain and age-at onset of NIDDM but is also associated with susceptibility to NIDDM. PMID- 8721783 TI - Intracerebroventricular administration of neuropeptide Y to normal rats increases obese gene expression in white adipose tissue. AB - The aim of this work was to determine the possible inter-relationship between neuropeptide Y (NPY, a hypothalamic stimulator of feeding) and adipose tissue expression of the ob protein (a novel potent inhibitor of feeding). Such a relationship could be of importance in the maintenance of normal body weight. To this end, normal rats were intracerebroventricularly (i.c.v.) infused for 6 days with NPY. NPY infusion resulted in hyperphagia and a marked increase in adipose tissue ob mRNA levels. The effect of NPY on ob expression persisted when hyperphagia was prevented by pair-feeding, and was reversed following cessation of NPY infusion. Basal and glucose-stimulated insulinaemia were increased by i.c.v. NPY infusion compared to control values, regardless of whether animals were ad libitum-fed or pair-fed. Cessation of NPY infusion was accompanied by normalisation of insulinaemia. These changes in insulinaemia produced by i.c.v. NPY infusion paralleled the observed changes in ob expression. When normal rats were made hyperinsulinaemic-euglycaemic for 24 h, such hyperinsulinaemia also resulted in increased ob mRNA levels in white adipose tissue. This suggested that NPY-induced hyperinsulinaemia could be responsible for the upregulation of ob mRNA levels of NPY-infused rats. It is concluded that central (i.c.v.) NPY infusion increases adipose tissue ob expression, a functional relationship that is linked, at least in part, via NPY-induced hyperinsulinaemia. PMID- 8721785 TI - The need for a 'free radical initiative'. PMID- 8721786 TI - Cholesterol-lowering therapy may retard the progression of diabetic nephropathy. PMID- 8721784 TI - Oxidative stress and insulin action: is there a relationship? PMID- 8721787 TI - The effect of cholesterol-lowering therapy on the progression of diabetic nephropathy is unproved. PMID- 8721788 TI - Normal values of first-phase insulin response to intravenous glucose in healthy Italian children and adolescents. PMID- 8721789 TI - Age-dependent association of HLA-A24 in Japanese IDDM patients. PMID- 8721790 TI - Report from the post-EASD symposium of the EASD-Islet Study Group, "Pancreatic Islet Research-Towards the Year 2000", held 16-18 September 1995, Sigtuna, Sweden. PMID- 8721791 TI - [Mechanism of DNA recognition by a protooncogene product, Myb]. PMID- 8721792 TI - [Structural studies of phorbol ester-binding domain of protein kinase C by photoaffinity labeling]. PMID- 8721793 TI - [Abnormality of cytoskeletons and their phosphorylation in Alzheimer's disease]. PMID- 8721794 TI - [YAC-modification system for analyzing gene regulation]. PMID- 8721795 TI - [Angiogenesis and enzymatic activity of thymidine phosphorylase]. PMID- 8721796 TI - [The structure of smooth muscle myosin phosphatase]. PMID- 8721797 TI - Assessing the quality of reports of randomized clinical trials: is blinding necessary? AB - It has been suggested that the quality of clinical trials should be assessed by blinded raters to limit the risk of introducing bias into meta-analyses and systematic reviews, and into the peer-review process. There is very little evidence in the literature to substantiate this. This study describes the development of an instrument to assess the quality of reports of randomized clinical trials (RCTs) in pain research and its use to determine the effect of rater blinding on the assessments of quality. A multidisciplinary panel of six judges produced an initial version of the instrument. Fourteen raters from three different backgrounds assessed the quality of 36 research reports in pain research, selected from three different samples. Seven were allocated randomly to perform the assessments under blind conditions. The final version of the instrument included three items. These items were scored consistently by all the raters regardless of background and could discriminate between reports from the different samples. Blind assessments produced significantly lower and more consistent scores than open assessments. The implications of this finding for systematic reviews, meta-analytic research and the peer-review process are discussed. PMID- 8721798 TI - Relationship between total mortality and cholesterol reduction as found by meta regression analysis of randomized cholesterol-lowering trials. AB - All randomized trials of cholesterol lowering except those without information on total mortality or cholesterol reduction were included in this meta-analysis. Its purpose was to review the relationship between log odds ratio of total mortality in active vs. control group and degree of cholesterol reduction, adjusting for appropriate covariates such as type of trial (single or multifactor), risk level in the control group, and type of treatment. Analysis revealed that risk level in the control group was particularly important to adjust for because it correlated both to degree of cholesterol reduction (r = 0.48) and to log OR of total mortality (r = -0.38, single-factor trials) and thus confounded the cholesterol mortality association. After adjustment, a borderline significant association was found between degree of absolute cholesterol reduction and total mortality log OR. Type of treatments also seemed to be associated with log OR, mostly through excess risk in hormone-treated patients. There was no statistically significant indication that fibrate trials had inferior results as compared to other drug trials. Meta-analysis estimates of global treatment effects can be misleading in the guidance for creating appropriate recommendations regarding cholesterol reduction. More emphasis must be put on how cholesterol is reduced and how much. PMID- 8721799 TI - Community volunteers as recruitment staff in a clinical trial: the systolic hypertension in the elderly program (SHEP) experience. AB - During the recruitment phase of a trial it is often found that recruitment is not proceeding as quickly as projected. Budget limitations require innovative methods, such as use of volunteers, to increase recruitment yields without increasing cost. In the Systolic Hypertension in the Elderly Program (SHEP), volunteer staff at 12 of the 16 clinical centers (CCs) performed a range of tasks such as mailings, telephoning, and clerical work. SHEP volunteers donated almost 40,000 hours, at an estimated cost savings to the program of more than $368,000. Staff volunteers appear to require more training and supervision than regular staff and may require flexible work schedules. These limitations can be obviated by careful planning. This paper describes the SHEP experience with staff volunteers and provides suggestions for their use in other trials. PMID- 8721800 TI - Maximizing recruitment efforts in a drug lipid-lowering trial with dietary intervention to lower LDL cholesterol. AB - A select group of screened applicants initially disqualified from a four-center, primary prevention drug lipid-lowering trial because of borderline elevated serum low-density lipoprotein cholesterol (LDL-C) levels, as defined in National Cholesterol Education Program-Adult Treatment Panel I (NCEP-ATP I) guidelines, participated in a dietary intervention protocol that was incorporated into the screening phase of the trial. Seventy-seven screened applicants for the Asymptomatic Carotid Artery Progression Study entered the dietary program, which was overseen by an experienced registered dietitian at the central operations sites who collaborated with local staff at clinical sites during program implementation. NCEP-ATP I fat-modified step I diet specifications served as the basis for the intervention. The program, consisting of five sessions conducted over an 8-week period, primarily used written and audiovisual educational materials in combination with behavioral approaches. Of the original 77 participants, 36 responded to the intervention by achieving their LDL-C goal. Twenty-nine were nonresponders and 12 were dropouts. Responders achieved an average 11.7% drop in total cholesterol at the end of the 8-week program. Mean LDL-C decline paralleled total cholesterol change. High-density lipoprotein cholesterol also decreased significantly. These results were sustained for 24 of the responders attending the final screening visit approximately a month later, when another fasting blood lipid measurement was made. Participants who dropped out were more likely to be smokers. Pre- and postintervention nutrition data assessed by semiquantitative food frequency questionnaire for 20 screenees randomized into the study indicated significant reductions in total fat, saturated fat, polyunsaturated fat, and dietary cholesterol, all known to influence blood lipid levels. Similar programs may prove useful to other drug lipid-lowering trials to maximize recruitment efforts. PMID- 8721801 TI - Clinical trial enrollers vs. nonenrollers: the Cardiac Arrhythmia Suppression Trial (CAST) Recruitment and Enrollment Assessment in Clinical Trials (REACT) project. AB - The Recruitment and Enrollment Assessment in Clinical Trials (REACT) was a National Heart, Lung, and Blood Institute (NHLBI)-sponsored substudy to the Cardiac Arrhythmia Suppression Trial (CAST). Two-hundred-sixty (260) patients who enrolled in CAST and 140 partially or fully eligible patients who did not enroll were compared across several parameters, including demographic variables, disease severity, psychosocial functioning, health beliefs, recruitment experience, and understanding of informed consent procedures used in CAST. Significant predictors of enrollment included several demographic variables (e.g., being male, not having medical insurance), episodes of ventricular tachycardia, and health beliefs (e.g., extra beats are harmful, a higher degree of general health concern). Enrollment was higher for those who read and understood the informed consent and those who were initially recruited after hospital discharge, particularly nondepressed patients. In the multivariate model, the key variables that emerged were the patient's reading of the informed consent form and the patient's lack of medical insurance. These results suggest that (1) the clinical trial staff's interaction with the patient and the time when recruitment is initiated contribute significantly to the decision to enroll; and (2) it may be a greater challenge to motivate patients to enroll in future clinical trials if health care reform improves access to medical insurance coverage. Some of the significant variables are modifiable, suggesting interventions that may increase enrollment rates in future trials. PMID- 8721803 TI - Do low observed error rates lessen the need for study-specific data quality audits? PMID- 8721802 TI - Is evening primrose oil of value in the treatment of premenstrual syndrome? AB - A systematic literature search of clinical trials of evening primrose oil (EPO) for the treatment of the premenstrual syndrome (PMS) was carried out with a view to performing a meta-analysis. Only seven placebo-controlled trials were found but only in five trials was randomization clearly indicated. Inconsistent scoring and response criteria made statistical pooling and hence a rigorous meta-analysis inappropriate. The two most well-controlled studies failed to show any beneficial effects for EPO, although because the trials were relatively small modest effects cannot be excluded. Nonetheless, on current evidence EPO is of little value in the management of premenstrual syndrome. PMID- 8721804 TI - Rationale, design, implementation, and baseline characteristics of patients in the DIG trial: a large, simple, long-term trial to evaluate the effect of digitalis on mortality in heart failure. AB - This article provides a detailed overview of the rationale for key aspects of the protocol of the Digitalis Investigation Group (DIG) trial. It also highlights unusual aspects of the study implementation and the baseline characteristics. The DIG trial is a large, simple, international placebo-controlled trial whose primary objective is to determine the effect of digoxin on all cause mortality in patients with clinical heart failure who are in sinus rhythm and whose ejection fraction is < or = 0.45. An ancillary study examines the effect in those with an ejection fraction > 0.45. Key aspects of the trial include the simplicity of the design, broad eligibility criteria, essential data collection, and inclusion of various types of centers. A total of 302 centers in the United States and Canada enrolled 7788 patients between February 1991 and September 1993. Follow-up continued until December 1995 with the results available in Spring 1996. PMID- 8721805 TI - Significance of programmed cell death (apoptosis) in mammalian limb and palate development. PMID- 8721806 TI - [Heterogenity of diacylglycerol Kinase--Structure, biochemical characteristics and gene expression in the brain]. PMID- 8721808 TI - A histocytological study of the rat hair follicle--an appraisal of a monocellular layer ("middle root sheath") between inner and outer root sheaths. AB - Hair follicles of male Wistar rats in the anagen stage were examined by light and electron microscopy. A distinct monocellular layer was identified between the Henle's layer and the outer root sheath (ORS). In the lower part of the ORS, this cell layer is more closely bound to the inner root sheath (IRS) than to the ORS, while in the upper half the situation is reversed. Electron density and staining property of this layer are gradually changed according to regions of the hair follicle which may indicate the metabolic conditions of the organism. The origin of this layer can be traced back to the germinal matrix directly facing the neck of the hair papilla. Findings are also reported about the gaps formed between the cells of Henle's layer after keratinization and about the contact formed between the cells of Huxley's layer and the cells of this monocellular layer through these gaps. Our observation reported in this study strongly suggests that this layer belongs neither to the IRS nor to the ORS, but that it represents an independent layer of cells. Therefore we propose to name this layer the "middle root sheath." This middle root sheath may serve the following three functions: 1) alleviation of intrinsic friction generated by colliding cell dynamics between the IRS and the ORS, 2) deterrence of keratinization of the ORS after exfoliation of the IRS and 3) secretion and excretion of metabolites. PMID- 8721807 TI - Expression of heparin-binding growth factors and their receptors in neural development. PMID- 8721809 TI - -Structure of the respiratory system of the Ulodera Hynobius neblosus tokyoensis Tago, with special reference to the distribution of serotonin-immunoreactive cells in its respiratory tract epithelium. AB - The respiratory tract epithelium in many vertebrate species has a neuroepithelial endocrine (NEE) system. This system is composed of solitary NEE cells or organoid cell groups called neuroepithelial bodies (NEBs). In response to chemical and physical stimuli, NEE cells may release bioactive substances. Serotonin, one of the biogenic amines, well-known as a constricter of smooth muscle, can be found in NEE cells, serotonin-immunoreactive cells can be used as a marker for these cells. Comparative histological studies of lower vertebrates can improve our understanding of mammalian respiratory systems. The Tokyo salamander (Hynobius neblosus tokyoensis Tago), classified as Ulodera, is particularly useful for comparative studies of respiration. In this study, the serial sections of respiratory tract of the Tokyo salamander were stained by a commonly used staining method and by an immunocytochemical method for serotonin, and the distribution of serotonin-immunoreactive cells in the respiratory tract was examined. The respiratory tract was found to be connected to the alimentary tract via an aditus laryngis, which opens on the medio-ventral side of the esophagus. The laryngotrachea was slit-like or elliptically shaped with a total length of about 3.5 mm, joining the aditus laryngis. The laryngotrachea was supported by a pair of lateral cartilages, and a fibromuscular layer was seen between the cartilages and the epithelium. In the cranial region, a laryngeal sphincter was seen around the laryngotrachea. The laryngotrachea branches into a pair of tube like lungs, that are about 17-20 mm in length. Two apposed primary trabeclae run along the entire length of the lung wall, perpendicular to the axis, and containing the pulmonary arteries and veins. The lungs were divided into two portions: 1) an airway portion (trabeclae, septa) in which smooth muscles surrounding the large vessels were well developed, and 2) a respiratory portion which was give that name because it has well developed capillary networks that were assumed to be involved in gas exchange. The lumen of the laryngotrachea and the pulmonary airway portion contained pseudostratified cilio-mucous epithelium. In the caudaldorsal region of the laryngotrachea adjacent to the lungs, the non ciliated respiratory epithelium was seen lining the capillaries. In cilio-mucous epithelium of the laryngotrachea, all serotonin-immunoreactive cells were solitary. They apposed to be columnar, cuboidal, triangular, oval, and flask- or spindle-shaped. Solitary serotonin-immunoreactive cells were classified "open type cell" with appical process reaching to the luminal surface and "closed type cell" insulated from the lumen by an epithelial lining. In the pulmonary airway portion, serotonin-immunoreactive cells were solitary cells and in clusters. Serotonin-immunoreactive cells were widely distributed throughout the respiratory tract, but they tended to be found mainly in the cranial portion. The density was highest in the area with the laryngeal sphincter, and decreased caudally in the laryngotrachea and lung. No serotonin-immunoreactive cells were found in the respiratory portion of the dorsal-caudal area of the laryngotrachea or in the part of the lung with non-ciliated cells. So the structure and distribution of serotonin-immunoreactive cells in the respiratory tract of the Tokyo salamander are similar to those of NEE cells and NEBs in mammalian respiratory systems. The density of serotonin-immunoreactive cells appears to be related to the distribution of smooth muscles in the fibromuscular layer and airway portion. The cells may be involved in regulation of the respiratory system. Serotonin is released in response to stimulation, which could result in constriction of the fibromuscular layer and shrinkage of the laryngotracheal cavity, and may regulate pulmonary volume by constricting smooth muscles PMID- 8721810 TI - [Examination on the courses of the arteries in the axillary region I. The course of the subscapular artery system, especially the relationships between the arteries and the posterior cord of the brachial plexus]. AB - The subscapular artery (Sbs) is the largest and most variable branch of the axillary artery (Ax). Although this Sbs has been defined as a common trunk of the thoracodorsal artery (TD) and the circumflex scapular artery (CS), TD and CS do not always form a common trunk, and the posterior circumflex humeral artery (CHP) is often associated with CS or the CS+TD trunk. Moreover, the subscapular branch (RS) usually arises from CS or CS+TD. Therefore, the subject of the study on Sbs must at least include RS and CHP. However, many anatomical studies on Sbs variation have not included them, and the mechanism causing the variation of Sbs has not been treated except in a few papers. Yamada distinguished two types of subscapular artery in 1967. The first type arose from Ax proximal to the point where Ax passed between the lateral and the medial cord or penetrated the ventral stratum of the brachial plexus. The second type arose from Ax distal to the point. The former type crossed over the medial cord of the plexus to reach the upper lateral thoracic wall, gave off the lateral thoracic artery (TL) and then ran backward on the serratus anterior muscle to reach the deep region of the axilla. Yamada named the former type Sbs "A.subscapularis superficialis" and considered it to be derived from TL. Yamada made an epoch-making report regarding the arteries of the axilla in 1967. He did not, however, refer to the relationship between the arteries to the deep region of the axilla and the posterior cord of the plexus. We called Sbs and the related arteries to the deep region of the axilla "the subscapular artery system" (Sbs system) and examined the courses of the Sbs system in 202 sides of cadavers during the dissecting practices from 1988 to 1992. Our examination revealed that there were three types of Sbs systems, i.e., S-type, I-type, and P-type Sbs systems. The S-type Sbs system was almost the same as Yamada's "A. subscapularis superficialis", branching off from the lateral thoracic artery arising from Ax proximal to the point where Ax penetrated the ventral stratum of the brachial plexus, and ended up in TD or bifurcating into TD and CS. CHP arose from the proximal part of the TD+CS-trunk and ran via the proximal course, or from CS and via the distal course. The S-type CHP ran posterior to the radial nerve (R). The frequency of appearance of the main arteries of the S-type Sbs system decreased sequentially from the medial artery to the lateral one as follows: TD > CS > CHP. Both the I type and P-type Sbs system arose from Ax distal to the point where Ax penetrated the ventral stratum of the brachial plexus. But the former ran posterior across the medial side of R and the latter ran posterior across the lateral side. The branching point of the I-type Sbs system from Ax was located more proximally than that of P-type. The I-type Sbs system more often gave off CS than TD and CHP. The P-type Sbs system arose from the most distal part of Ax just in front of the lateral axillary hiatus. The sequence of the frequency of appearance was CHP > CS > TD. Both CS and TD of P-type passed behind R. Different types of Sbs systems coexisted. In such cases, the more proximal type gave off more medial arteries and the more distal type gave off more lateral arteries. Exceptions to the rule, for example, coexistence of P-type CS with S-type CHP or I-type TD with S-type CS etc., were never observed. Although different features were distinguished in the stem parts of the three types of Sbs systems, the courses of the peripheral branches of Sbs systems displayed a common pattern on the surface of the subscapular muscle when they were analyzed collectively disregarding the direction of the flow of blood. Therefore, the peripheral part of each Sbs system on the surface of the subscapular muscle was considered to be derived from a common arterial network. In conclusion, the mechanism of the formation of variations in the Sbs system can be understood by the combination PMID- 8721811 TI - Vincula tendinum of human hands with special reference to vascular patency. AB - As persisting parts of the mesotendon, Vinculum breve and Vinculum longum are known to serve as transport and conduction pathways to the intravaginal segments of flexor tendons. For morphological evaluation of their constancy, they were dissected out of, measured, and histologically examined on the 2nd-5th fingers in 53 hands of donated cadavers. The Vincula brevia existed in all cases more or less in association with the insertion of the superficialis as well as profundus flexor tendon, meanwhile the Vincula longa varied in number from 0 to 3 per tendon from one finger to the others, even 4 in the case of the index superficialis tendon. Light microscopic investigation of vascular patency was carried out on semithin resin sections of 137 Vincula longa obtained from the flexor superficialis and profundus tendons of 53 index fingers. Up to 5 or more arterial and venous vessels were counted per Vinculum longum. The complete absence of a patent artery was recognized in 6 out of 62 Vincula longa of the flexor superficialis and in 22 out of 75 of the profundus flexor. Most frequently, the Vincula longa had a single artery and a single vein. Considering the constant presence of vessels on the cross section, the Vinculum breve is regarded as essential for maintaining the tendons at work. In contrast, the significance of Vinculum longum will be variable for the microcirculation of intravaginal segments of the flexor tendons individually and also in the course of aging. PMID- 8721812 TI - Persistent median artery: an anatomical study in neonatal and adult cadavers. AB - To examine the association of the median artery with the median nerve, 110 forearms from 20 neonatal and 35 adult cadavers of Turkish populations were dissected and compared with those of adult cadavers. In neonatal cadavers, the persistent median artery was found unilaterally in 5 (12.5%) forearms, in 4 of which the artery arose from the ulnar artery and in one from the common interosseous artery. In adult cadavers, however, no persistent median artery was observed. From these results, it could be suggested that this artery undergoes regression even after birth. PMID- 8721813 TI - Axonal transport of protein products of the bacterial reporter gene in the brain of transgenic mice. AB - The aim of the present study was to examine the intracellular localization of ectopically expressed proteins of the bacterial reporter gene in neurons of transgenic mice. In the brain of transgenic mice that carry a chimeric gene composed of the human tyrosine hydroxylase gene promoter and the bacterial gene encoding chloramphenicol acetyltransferase (CAT), the expressed CAT protein was previously found in catecholaminergic neurons as well as in non-catecholaminergic neurons expressing CAT ectopically. In the gene-expressing catecholaminergic neurons, the CAT protein was formerly detected in the axon terminals. In the present study, we immunocytochemically showed that the ectopically expressed protein was localized in cell bodies and axon terminals of the granule cells of the dentate gyrus. This suggests that the ectopically expressed protein of the bacterial gene is transported in the axons to their terminals. PMID- 8721814 TI - [A territorial profile of tuberculosis in Romania in 1994 with details on the location of the disease, the bacteriological confirmation of pulmonary forms and the grouping of patients based on age and sex]. PMID- 8721816 TI - [Tuberculosis in the city of Bucharest in 1989-1993]. AB - The global incidence of tuberculosis increased during the last 5 years in the city of Bucharest by 37.8%, from 84.2%000 in 1989 to 116.0%000 in 1993. The risk of disease was permanently higher in the rural area of the Ilfov Agricultural Sector (148.5%000 in 1993). During 1993 almost 55% of new cases of pulmonary tuberculosis registered in the city of Bucharest presented advanced lesions on case-finding; 71% of these cases were tb bacilli eliminators. In the epidemiological conditions induced by a marked increase of contamination sources (periodic prevalence of tb bacilli eliminators reached 111.6%000 in 1993) the disease risk in children increased as well in Bucharest from 19.6%000 in 1989 to 38.3%000 in 1993. Tuberculosis control in Bucharest benefits from a rather complex material basis which should be improved according to the present day epidemiological conditions. PMID- 8721815 TI - [The incidence of tuberculosis in Romania in 1994]. AB - The 1994 TB notification rate structure in Romania was based on the data reported in "New-case/relapse TB notification form", registered between 1.01.-31.12.1994 and stored in "National TB-Register" data base. The 21,422 TB patients notified in 1994 correspond to an annual incidence of 94.2%000, which means an increase of 5.41% as compared with 1993, of 34.6% as compared to 1990 and of 68.8% as compared to 1985. As compared with 1985 tb notifications, this increase supplied a commulative overload of 32,546 patients, out of which, 18,233 only in the last 5 years. Tb notification rate show large differences between districts, varying from 38.9%000 (Harghita district) to 130.2%000 (Giurgiu district). The great variations between the number of the quarterly reported cases for 1994 in different counties, having an alleatory distribution of the extreme values in different periods of the year, could represent an under-diagnosis. This hypothesis could be sustained also by the large differences in the ratio of the pleural tb effusions in various districts (from 16% of the TB-incidence in Brasov district to 1.4% in Satu Mare county) or the ratio of the other extra-pulmonary tb forms (that are dispersed between 1.5% in Salaj county to 8% in Constanta county). 3,428 patients were registered more than 3 months after the start of the treatment, 500 after more than 6 months and even 24 patients were notified more than 1 year later. Taking into account this delay in reporting and the facts already mentioned, we could suppose a higher number of cases than reported for a given time interval. The ratio of bacteriologicaly prouved pulmonary-TB cases was 76.4% (i.e. 61.6%000), varying in different counties between 48.4% (Arges) and 95.4% (Sibiu). These variations could rather be explained by the quality of the sputum-examination than by the profile of the registered patients. A large proportion (81%) of the confirmed cases are smear-positive, varying between 47.6% (Arges) and 88.6% (Bistrita). The high percentage of the smear-positive cases underscore the large number of "sources" whereas variations between the ratios are perhaps due to the quality of the smear examination. Ratio of pulmonary cases confirmed only by cultures (14.5% in Romania) revealed also teritorial differences, on a scale between 2.9% (Teleorman) and 34.5% (Botosani). To the number of the smear-positive cases notified during 1994, another 6,136 chronic patients (at the end of 1994) had to be considered. The peak prevalence of sputum positiv cases in 1994 (positive cases at the beginning of the year, plus positive cases added during 1994) was of 24,568 (108%000 inhabitants, 137%000 adults). New notified bacteriologicaly positive TB-cases (13,914) added to all chronic TB cases (6,136) reported, represent a lower number than "maximal bacilli excretors prevalance" reported. The lacking 4,500 positive patients could represent either cases reported as having no culture results (the culture results could have been registered two months later) or unreported cases. This situation could be corrected by a "double-reporting" with matching of the reports, starting in 1995. The TB notification rate in children was 19.8%000 (949 cases). The 2,405 tb deaths correspond to a TB-mortality of 10.6%000 in 1994. During 1994, 66,779 patients were hospitalised with "pulmonary tb" diagnosis, 44,915 (67.3%) were repeatedly negatives and 1,545 had no examination for mycobacteria reported. Out of the total of 20,248 positive cases, 15,398 became negative whereas 4,834 (23.9%) were released sputum-positive. The number and notification rate of tb cases by sex and age groups, localisation and bacterial confirmation (for pulmonary TB), by districts, are presented in tables and graphs (drown on the same scale), printed as supplement of this issue. PMID- 8721817 TI - [The treatment of bronchial asthma with powdered salbutamol and powdered beclomethasone dipropionate]. AB - The active bronchodilators and anti-inflammatory drugs used as dry powder in inhalatory treatment of bronchial asthma represent an important advance during last years. In this way the inconveniences of pressurized aerosols treatment where the drugs are only partially inhaled as well as severe side-effects of systemic corticotherapy are avoided. The bronchodilator drug Ventodisk and the antiinflammatory Becodisks as dry powder are given by inhalation with a special device (Diskhaler). The study deals with a group of 60 subjects with moderate and severe forms of bronchial asthma out of which 20 were treated by Ventodisk associated to Becodisks and other two groups of 20 subjects each treated during the same interval time with only Becodisks and Ventodisk respectively. The clinical results after two months treatment were much better in the first group of subjects (85% good results) in comparison to Becodisks (75%) and Ventodisk (60%) treated cases. One can state in conclusion that the association of Ventodisk to Becodisks leads to very good results in basic treatment of bronchial asthma. PMID- 8721818 TI - [The role of the epidemiological survey in childhood tuberculosis]. AB - All epidemiological inquiries in tuberculous children are centralized and processed in the Institute of Pneumophtisiology. The analysis of 4,171 inquiries processed during 1985-1992 shows: after 1985 an increase of diseased children is noted. This increase is higher in gipsy children, whose weight reached 38.7% out of the total in 1992; the tb. children were found out mainly by symptoms and in 20-25% by ascendent epidemiological inquiry; the bacillary sources which infected the children have been found out by ascendent epidemiological inquiry in 70% of cases; the infection sources consisted mainly in new lung tb. cases in adults (78 82%), relapses (6-8%) and chronic cases (12-14%). In certain areas, the weight of chronic cases as infection sources exceded 20%. PMID- 8721819 TI - [A profile of the smoker with pulmonary tuberculosis]. AB - A group of 485 subjects suffering from lung tb admitted in the Institute of Pneumophthisiology "M. Nasta" has been studied as for the relationships between found the lung disease and smoking. High frequency of smoking has been among these patients: 87.7% in males and 68.2% in females. These figures marquedly exceed the frequency of smoking in the whole population. The increase of frequency among females is more evident. No difference has been found as for the type of lesions between smokers and non-smokers. Smoking increases the gravity of associated diseases (e.g. chronic ethylism) evidencing the lack of health education of people in preventing this condition as well as the difficulties met by physicians in the implementation of smoking abandonment programs. PMID- 8721820 TI - [Functional study of the respiratory muscles in patients with diffuse interstitial pneumopathies (DIP)]. AB - The aim of the study was to evaluate the function of the respiratory muscles in patients with diffuse interstitial lung diseases (DILD) by measuring the maximal static inspiratory (MIPS) and expiratory (MEPS) pressures and to find out a possible relationship between these pressures and some parameters of the respiratory function of the lung. The battery of functional tests consisted in: lung volumes, flow rates, lung elasticity and arterial partial pressure of oxygen at rest and on exercise. MIPS and MEPS were measured using the Black and Hyatt method. 82 patients with DILD of different etiologies were studied, 39 males and 43 females, with a mean age of 33.6 +/- 8.74 years. The results signaled the decrease of MIPS in 29 cases--35.3% (the mean value in % predicted and the standard deviation: 47.0 +/- 9.75) and of MEPS in 47 cases--57.3% (the mean value in % predicted and the standard deviation 49.7 +/- 9.3). No relationship has been found between the low values of MIPS and MEPS on one hand and the altered values of some parameters of the lung function (diminished lung volumes, increased elastic recoil, hypoxemia on exercise) on the other. The peak expiratory flow rate--PEF--was significantly greater in patients with normal values of MIPS and MEPS than in those with decreased ones, which confirms the dependence of PEF on the force of the respiratory muscles. In our study, despite previous published data, the decrease of the force of respiratory muscles in patients with DILD seems to appear with a rather high frequency. PMID- 8721821 TI - [The clenbuterol treatment of patients with the chronic obstructive syndrome]. AB - We have studied the effect of a new long acting bronchodilator: CLENBUTEROL, in 10 patients suffering from chronic obstructive diseases. All patients received 1 tablet of Clenbuterol, then we measured the following parameters in each patient, 2 and 6 hours after the drug administration: VC (vital capacity), FEV1 (forced expiratory volume in the first second of forced expiration), FEV1 x 100/VC, Raw (airways resistance), RV (residual volume). In all patients, excepting one of them, we registered a modification of all or some of the parameters 2 hours after drug administration. The bronchodilator effect was still present at 6 hours after drug administration. This data suggest that Clenbuterol induced a significant and long acting improvement of chronic obstruction; this drug might offer the advantage of sustained effect in patients suffering from chronic bronchospastic obstruction. PMID- 8721822 TI - Pulmonary mechanics in patients with asthma bronchiale in symptom-free interval. AB - Pulmonary and airway mechanics were assessed in 30 patients with asthma bronchiale during a symptom-free interval, and with normal spirometry. Static expiratory pressure-volume (PV) curves and maximum expiratory flow-volume curves were obtained, and maximum flow-static recoil (MFSR) curves were drawn-up. In 11 patients elastic pressures were significantly lower than normal at all lung volumes, but the slope of the PV curve was normal; the loss in lung elastic recoil was not associated with an increase in lung volume, suggesting that surface forces acting in the alveoli may be responsible for the elastic abnormality. In 5 patients static pressures were increased at all lung volumes, possibly due to the increase in elastic pull exerted by the thorax on the lungs. Obstruction of small airways was present in most cases: MEF50 was lower than predicted in 70% of them, and in 73.3% the upstream resistance was increased. In 17 subjects, the MFSR curves showed increased pressures at given maximal flows suggesting that the airways obstruction was generated by an intrinsic mechanism. In 4 cases, an extrinsic mechanism might be responsible since the elastic pressures at given maximal flows were lower than normal. PMID- 8721823 TI - [The etiopathogenesis of bronchial asthma]. PMID- 8721825 TI - [Goodpasture's syndrome--a case report]. AB - A female subject case aged 64 years is presented. The history shows several hospital admissions for lung diffuse infiltrative processes, accompanied with hemoptoic sputa. These infiltrations had a regressive evolution either spontaneously or under treatment. The diagnosis of Goodpasture syndrome was fixed post-mortem by alterations evidenced in the kidneys. PMID- 8721824 TI - [Pulmonary manifestations in a case of toxic shock syndrome]. AB - The evolution of a toxic shock syndrome in a young adult has yielded particular pleural and lung lesions concomittantly with phenomena of insufficiency of other organs and systems (kidney, digestive, CNS a.o.). X-ray aspect of bilateral bronchopneumonia with small diffuse infiltrates are noted, together with an aspect of bilateral spontaneous pneumothorax, which appeared successively with a tendency to relapse. Hemodynamic alterings during the acute phase of the syndrome initiated by staphylococcal exotoxin are similar to those during the initial phase of the septic shock, requiring a careful monitoring during the treatment. PMID- 8721827 TI - [Apropos of tuberculosis]. PMID- 8721826 TI - [Pulmonary cryptococcosis--a pathology of the future?]. PMID- 8721828 TI - [The 9th World Congress on Smoking or Health, Paris, 9-14 October 1994]. PMID- 8721829 TI - [Smoking in rural areas in Romania]. PMID- 8721830 TI - [Amantadine in the prevention of nosocomial influenza]. AB - Amantadine prophylaxis was performed in 91 patients during influenza A epidemics. It was used for patients with chronic heart, pulmonary and metabolic disease, for immundeficienty and elderly patients. Patients with gravidity, lactation, epilepsy, peptid ulcer or serious liver disorder were excluded from prophylaxis. The daily dose was 200 mg, which was reduced to 100 mg in people over 65. The chemoprophylaxis was combined with killed influenza vaccine in 6 patients. No influenza-like illness occurred among patients with prevention. Light side-effect was observed in 5 patients. Ten peoples who were excluded from prophylaxis caught serologic proven influenza. Amantadine prophylaxis is appropriate for prevention of nosocomial influenza among high-risk patients in institutions because of other diseases. PMID- 8721831 TI - [Atopy score in infancy and the severity of asthma at 7 years of age]. AB - Due to limitations in early diagnosis of atopy at wheezy infants in 1982 the authors introduced a special scoring system called "atopy-score". This evaluates the skin prick tests' results, serum IgE level, the presence of other allergic symptoms, and atopy in the family history. The relationship between atopy-score in early childhood and severity of atopic asthma at age 7 was investigated in 30 children (15 mild, and 15 severe asthmatics). It was proved, that atopy-score during infancy was high in those children who had severe atopic asthma at seven, and low in those who developed mild disease: 9.2 vs. 5.9. The findings indicate that wheezy infants with high atopy-score are candidates for preventive measures and early protective treatment. PMID- 8721832 TI - [Incidence of spontaneous bacterial peritonitis in cirrhotic patients with ascites. A two-year prospective study]. AB - The prevalence of spontaneous bacterial peritonitis in patients with decompensated liver cirrhosis admitted to our unit during last two years was studied. Criteria of spontaneous bacterial peritonitis were positive bacteriology and greater than 250/mm3 polymorphonuclear leukocyte count. 84 examinations in 50 patients were made. Spontaneous bacterial peritonitis was diagnosed in 4 cases. E. coli, Klebsiella pneumoniae, alpha haemolyzing Streptococcus were isolated from ascitic fluid in positive cases. Three patients died despite of therapy, one patient recovered. Bacterascites (positive bacteriology without increased polymorphonuclear leukocyte count) was detected in two cases. Culture-negativ neutrocytic ascites (greater than 250/mm3 polymorphonuclear leukocyte count without positive bacteriology) was detected in four cases. The clinical picture of above-mentioned cases was symptomless. Low protein concentration of ascitic fluid (less than 10 g/l) predisposing to spontaneous bacterial peritonitis, was found in 15% of cases. Incidence of spontaneous bacterial peritonitis and equivocal states found by us was similar to the data given by the literature: 15 20%. Because of the high mortality rate and frequent symptomless course in cirrhotics, importance of diagnostic paracentesis is stressed. PMID- 8721833 TI - [Low-molecular-weight heparin in the prevention of thromboembolism in pregnant thrombophilic patients]. AB - Prophylaxis of thromboembolism with low molecular weight heparin (LMWH) may offer some advantages over unfractionated heparin during pregnancy. Controlled studies with LMWH for thromboprophylaxis in pregnancy are failing, although according to some recent studies LMWH did not cross the placental barrier. LMWH as thromboprophylactic agent was used in three young pregnant women with familial thrombophilia (two with PC one with AT-III deficiency). According to the bodyweight of patients the applied doses of LMWH were 5,000-10,000 ICU once daily. Laboratory control (determination of anti-FXa activity in plasma samples) was made monthly. The three pregnancies were uneventful, thromboembolic or haemorrhagic complications did not develop. Newborns were healthy, with no coincide of disturbances of haemostasis. The LMWH-demand is certainly increased at the late period of pregnancy. LMWH as thromboprophylaxis is recommended for pregnants with familial thrombophilia. The necessary dose of Fraxiparine may be 70 ICU/kg/day in the first and 100 ICU/kg/day in the second half of pregnancy. PMID- 8721835 TI - [Russian witnesses to the Hungarian cholera epidemic 1848/49]. PMID- 8721834 TI - [Intrauterine ultrasonic diagnosis of ectopia cordis associated with exencephaly]. AB - The heart positioned outside the chest is called ectopic heart. The forms of ectopia cordis is generally a particular sign of a multiplex malformation and/or genetic aberration. According to the types of this malformation the authors described the sonographic signs, incidence and consequences of the condition on the base of a clinical case. PMID- 8721836 TI - [Vilmos Lippich, forgotten pioneer of medical statistics and fight against alcoholism. Addition to footnotes]. PMID- 8721837 TI - [The changing picture of rheumatic fever and the permanent need for its prevention]. PMID- 8721838 TI - [Radiation-induced cytogenetic markers detected 8 years after the accident at the Chernobyl Atomic Electric Power Station by different methods of analyzing metaphase chromosome preparations in persons who have had acute radiation sickness]. AB - A comparative cytogenetic observation of 10 patients suffered from acute radiation sickness of second and third degree as a result of Chernobyl accident has been carried out. The new data about the level of unstable and stable biomarkers of irradiation delayed exposure were established using conventional G banding and FISH-staining. PMID- 8721839 TI - [The distribution of chromosome aberrations in cells under the action of radiation of different qualities]. AB - The distribution of chromosome aberrations in the cultured human peripheral blood lymphocytes exposed to 6 MeV neutrons was compared with that of lymphocytes exposed to gamma-rays. The regularities of both distributions were similar. PMID- 8721840 TI - [A cytogenetic assessment of the mutagenic background in the Dnieper River industrial area]. AB - Cytogenetic investigation of plant cells and somatic cells of children from Dnepropetrovsk shown and increase of the frequency of chromosome aberrations and gene mutations in contaminated regions that testified the increased mutagenic background of urban environment. PMID- 8721841 TI - [The phenomenon of premature chromatid disjunction in the tissues of human embryos in vivo, in vitro and in pregnant women]. AB - The study of cell karyotype features in human medical abortions and tissues of pregnant women indicated the phenomenon of premature chromatid (centromere) disjunction. The association of the frequency of premature chromatid disjunction with embryo sex and with the prolonged cell cultivation in vitro was revealed. PMID- 8721842 TI - [The mutagenic activity of suspended particles in the atmosphere in the industrial cities of Ukraine]. AB - The total mutagenic activity of chemical pollutants was studied by semi quantitative Ames test in atmosphere of towns with different specificity of industrial development: towns with developed metallurgy, chemical industry, and relatively clean towns. The air samples from towns of metallurgical and chemical industry shown middle mutagenicity. PMID- 8721843 TI - [An experimental assessment of the genetic consequences of anthropogenic environmental pollution by a complex of chemical industry factors]. AB - Tests for dominant lethal mutations and gonadotoxicity were carried out on 45 males of albino laboratory mice exposed during two months to unfavourable environmental factors. An increase of the level of dominant lethal mutations and pronounced gonadotoxic effects were detected in experimental groups. PMID- 8721844 TI - [Nucleolus organizers in the epithelium of serous ovarian tumors]. AB - Morphofunctional state of nucleolar organizers in epithelial cells of benign, and malignant serous ovarian tumours has been studied. A rise in the activity of nucleolar organizers depending upon an increase of the level of malignancy has been revealed. PMID- 8721845 TI - [An analysis of human marker chromosomes originating from chromosome 21 by using in situ hybridization]. AB - Four cases of patients with additional marker chromosomes are described. The clinical, cytogenetic and molecular cytogenetic methods have been used for investigation. The identification of marker chromosomes was made by using in situ hybridization and the collection of chromosome-specific DNA probes. All marker chromosomes were determined as originated from chromosome 21 with break-points in the region between 21q11 and 21q22. PMID- 8721846 TI - [The modifying action of a complex of physiologically active substances isolated from the human amnion on chromosome damage]. AB - It was determined that the complex of physiological active substances separated from human amnion reduced the frequency of the hepatocytes with chromosomal aberrations in rats. This reduction of chromosomal damages was noted at spontaneous and induced mutagenesis. The value of cytogenetical effect, the availability of raw material and the natural origin of the drug allow to propose it for further study and use as modifier of mutagenic action. PMID- 8721847 TI - [The cytogenetic analysis of unfertilized human oocytes]. AB - Cytogenetic analysis of 186 unfertilized human oocytes obtained from 145 women were carried out according to programs of in vitro fertilization. The ratio of different cytogenetic types of oocytes in their heterogenic population was determined. At least 30% of oocytes were shown to have nuclear and cytoplasmic immaturity. The level of chromosome anomalies in oocytes of second meiotic metaphase comprised 30.8%. PMID- 8721848 TI - [Segregation and genetic dispersion analysis of the pedigrees of breast cancer patients in the Kiev region]. AB - Complex clinical-genealogical and genetic study of 261 patients with breast cancer (BC) was carried out in Kiev region. It was demonstrated that the multifactorial origin of BC was of first importance in this population. The share of genetic component in the susceptibility to BC comprise 55.68 +/- 2.44%. Risk of the origin of malignant tumors in progeny was calculated. PMID- 8721849 TI - [Erythroid differentiation and specific protein synthesis by the leukemic cell line K-562 (under in-vivo cultivation conditions)]. AB - Prolonged cultivation of K-562 human leukemic cells in plastagar diffusion chambers inserted into the peritoneal cavities of CBA mice indicated erythroid commitment and differentiation of cells. It was shown by the method of dot hybridization with the use of oligonucleotide probe the presence of mRNA of globin in these cells. PMID- 8721850 TI - [A systems analysis of the relationships between phenotypic traits and the clinical polymorphism of respiratory diseases in children]. AB - Genetic phenotypic risk factors have been investigated in 1630 children with various forms of acute respiratory diseases and in control groups (314 children with bronchial asthma, 768 healthy children). Each of them was estimated by points using special elaborated prognostic tables. This allowed to distribute children into risk groups according to the development of disease from the moment of birth. PMID- 8721851 TI - [The formation of risk groups and the performance of invasive prenatal diagnosis]. AB - The results of the formation of women risk groups for invasive methods of prenatal diagnosis and cytogenetic investigations of amniotic fluid are presented. It was shown that the efficiency of prenatal diagnosis was increased due to the complex use of ultrasonography, screening for alpha-fetoprotein and chorionic gonadotropin, and invasive methods of prenatal diagnosis. PMID- 8721852 TI - [An analysis of the causes for the occurrence of Down's syndrome and the current approaches to its prevention at the stage of mass prenatal ultrasonic and immunobiochemical screening of pregnant women]. AB - Problems of prenatal complex diagnosis of Down's syndrome are considered. The levels of alpha-fetoprotein in blood serum of pregnant women should be studied in the optimum terms (17-19 weeks of pregnancy). The assessment of several immuno biochemical markers is most important for diagnosis. PMID- 8721853 TI - Methicillin-resistant Staphylococcus aureus (MRSA). PMID- 8721855 TI - Cerebrospinal meningitis: Nigeria. PMID- 8721854 TI - Cholera in Africa. PMID- 8721856 TI - [Geographic distribution of mortality from myocardial infarct in Baranya County]. AB - The acute myocardial infarction is one of the leading causes of death in Hungary. The geographical epidemiology is able to quantify the possibilities of preventive activities and to ascertain the target groups for intervention. On the basis of the analysis of standardized data from Baranya county, the mortality rate is 3.16 times higher in the settlements of the lower quartile compared to the rate in the settlements or higher quartile. This relative risk is equivalent with 68% of etiologic fraction. The results demonstrate that there are a group of settlements where local factors predominate the etiologic background of the myocardial infarction. The maps determine the actual clusters calling for some intervention. It seems to be very urgent task for the Hungarian public health organizations to fit the routine cluster investigation into the general public health practice since there is a definite demand for doing it. PMID- 8721857 TI - [Evaluation of complaints and diagnostic methods, considered to be objective, in gastroesophageal reflux]. AB - Authors analyse complaints of 166 patients suffering in gastroesophageal reflux disease, and results of different objective diagnostic methods (X-ray swallowing, endoscopy, 24 hour pH-monitoring, esophageal manometry). The degree of severity of typical complaints and changes found by diagnostic procedures were scored. Individual diagnostic proceduresindicated the characteristic changes of gastroesophageal reflux with great deviations. Hiatal hernia identified by x-ray, esophagitis observed by endoscopy, abnormal reflux revealed by pH-metry together occurred only at 36% of patients. Significant correlation could be revealed only between the degree of changes found by x-ray or pH-metry--applying them one by one--and the complaints of the patients, but with the combination of four diagnostic methods a tight, positive correlation could be demonstrated with the severity of the complaints of the patients in every case. Applying of diagnostic methods as much as possible completing each others, with consideration to the complaints of the patients is suggested for having the diagnosis been precise of the gastroesophageal reflux disease, and for measuring of efficacy of treatment. The importance of tight collaboration of internist, radiologist and surgeon and the significance of complex examination is underlined. PMID- 8721858 TI - [Serum elastin peptide concentration and human leukocyte elastase/antiproteinase balance in peripheral obstructive atherosclerosis]. AB - In some pathological states such as therosclerosis tissue destruction may be accelerated due to uncontrolled protease release of polymorphonuclear leukocytes and other events such as decreased concentration and/or the inactivation of main protease inhibitor molecules in the serum. In this study, the authors measured the elastase release of polymorphonuclear leukocytes which increased in atherosclerosis independently of the patients aged compared to healthy young subjects. These findings were similar to the response of polymorphonuclear leukocytes separated from healthy elderly subjects. Simultaneously, the main plasma proteinase inhibitors such as alpha-1-antitrypsin and alpha-2 macroglobulin in healthy and atherosclerotic subjects were determined. alpha-1 antitrypsin did not decrease significantly, whereas alpha-2-macroglobulin did in sera of atherosclerotic patients compared to age matched subjects (p < 0.05). In contrast, the activity of porcine pancreatic elastase was more effectively neutralized by the plasma obtained from healthy subjects suggesting diminished antiprotease activity of sera obtained from patients. The authors concluded that increased elastase release and decreased antiproteinase activity should be considered in atherosclerotic arterial wall damage. The similarity of the results in aged and therosclerotic subjects suggests that arteriosclerosis is an earlier aging process. PMID- 8721859 TI - [Chemical castration in homosexual pedophilia]. AB - The treatment of reducing sexual drive with antiandrogenic cyproterone acetate in a man with homosexual paedophilia is shown with the summary of rules for the use of cyproterone acetate in the therapy of sexual deviations. PMID- 8721860 TI - [Polycytemia vera in an 11-year old girl]. AB - The case of an 11 year old girl with three line type of polycythaemia vera, with 4 cm splenomegaly and a plethoric complexion is presented. Peripheral blood values were as follows: RBC: 7.75 x 10(12)/l, Hb: 18.8 g/l, WBC: 15.2 x 10(9)/l, platelets: 920 x 10(9)/l. Serum erythropoietin level: < 1 mU/ml. In vitro, erythroid colonies developed from the bone marrow in the absence of added erythropoetin. For three years the haematocrit value has been controlled by regular venesections. Since extreme thrombocytosis developed, the treatment was continued with interferon alpha. Different treatment protocols are discussed. PMID- 8721861 TI - [Hippocrates or Taigetos--ethical dilemmas in neonatal surgery]. AB - As a result of the decrease of the medical authority and the increasing power of the lawyers, the relationship between doctors and patients, patients and society has changed. The situation is more complicated in the case of neonates, infants and children who are unable to articulate their rights and wishes. The problem is complex in connection with the fetus because their interest and that of the mothers are often in conflict. The neonatal surgeon--only together with the geneticist, obstetrician and neonatologist--plays and important role in the evaluation of the mentally or somatically handicapped fetuses and neonates. The neonatal surgeon should suggest the termination of a pregnancy, should give his/her expert opinion about the result of the treatment of a malformation or should propose the withdrawal of the intensive care in the case of a critically ill neonate. Fetuses and neonates with congenital anomalies can be divided into 6 groups. 1. potential for total recovery--2. anomaly enabling a nearly normal life -3. malformations requiring permanent supervision and/or medical care--4. somatic rest defect and subnormal mental development--5. serious somatic and mental damage--6. anomal incompatible with life. In the complex evaluation of a fetus or a neonate with congenital malformation the neonatal surgeon--while not forgetting his/her Hippocratic oath--should take into consideration the quantity and quality of the rescued life, possibilities of a long treatment he/she should act according to the law and fulfil the expectations of the society. PMID- 8721862 TI - [Eosinophil activation markers in status asthmaticus]. AB - Eosinophil activation markers-eosinophil cationic protein, eosinophil protein-X were investigated in acute severe asthma. The elevation of the eosinophil activity markers were found in every cases at the time of admission. The measurements were repeated eight days later. The eosinophil cationic protein and eosinophil protein-X levels were increased at the time of the admission, and a decreasing tendency was detected eight days later. The peak expiratory flow rates were monitored during the period of the hospital staying. It is important to note that, the size of the bronchial obstruction did not show any similarity in all cases comparing to the elevation or decrease of eosinophil activity markers. The authors proved the eosinophil activation in vivo in acute severe asthma. The continuous monitor of the eosinophil activity markers might have a practical value in searching of the optimal therapeutic modalities in the asthmatic patients. PMID- 8721863 TI - [Cholesterol levels in young men and women planning conception]. AB - The purpose of this study is to incorporate the primary prevention of coronary heart disease into a periconception care. Among others total cholesterol was determined in 2610 female and 2307 male participants. The mean (+/- S.D.) of total cholesterol was 4.93 +/- 1.04 in females and 5.20 +/- 1.24 mmol/l in males. Only 7.9% of females and 4.7% of males had previous knowledge about their high total cholesterol. Three month later 1.08 mmol/l (16%) and 1.18 mmol/l (18%) reduction was found in total cholesterol of participants with > 6.5 mmol/l due to education programme including change in diet, to stop smoking, etc. PMID- 8721864 TI - [Analysis of costs of drug therapy in patients with acute respiratory insufficiency]. AB - In a retrospective study, the authors analysed the clinical data of 38 patients who were admitted to a surgical intensive care unit (SICU) for mechanical ventilation lasted for at least 72 hours. The APACHE III score was calculated on the basis of clinical data documented during the first 24 hour of the treatment and the cost of drug administrations per patients per day was also determined by analysing all the drugs prescribed on the first 5 days of intensive care. The patients admitted to SICU with polytrauma or with abdominal septic focus required significantly higher cost of drug treatment than the patients after cardiopulmonary resuscitation or with bronchial asthma or pneumonia. The APACHE III score of the patients died at the SICU was significantly higher compared to the survivors who needed more expensive drug therapy than the non-survivors required. Antibiotic treatment, blood transfusions, and human plasma proteins caused the highest drug expenditure. There was no significant correlation between the APACHE III score and the cost of drug treatment. PMID- 8721865 TI - [Inferior vena cava thrombosis developing in primary antiphospholipid syndrome]. AB - Antiphospholipid antibodies predispose to venous and arterial thrombosis. The case of a sixteen-year-old boy with primary antiphospholipid syndrome is presented. Features of different organ involvement such as thrombosis of the inferior vena cava, pulmonary thromboembolism and cerebral involvement were present with thrombocytopenia and high titre of anticardiolipin in the patient's sera. On twelve-month follow-up while taking anticoagulant the patient was entirely free of symptomes though certain immunological findings showed slight positivity. However, considering the ARA criteria systhemic immunopathologic disorder could not have been proved. Other pathological conditions with anticardiolipin positivity were also excluded. PMID- 8721866 TI - [Helicobacter pylori detection]. PMID- 8721867 TI - [Association of neurofibromatosis and carcinoid tumor]. PMID- 8721868 TI - [Medical fees from the radiologist's viewpoint (satisfaction instead of gratitude)]. PMID- 8721869 TI - [Correlation between p-53 expression and clinical resistance in testicular cancer]. AB - One of the most common cellular gene which negatively regulates the cell cycle, thus functioning as tumour suppressor gene, is the p-53 gene. The presence of this mutated gene has been correlated with, the aggressiveness of several malignant neoplasmas. Expression of the p-53 gene product protein was screened in 55 untreated human germ cell testicular tumours, furthermore a relationship between p-53 expression and clinical resistance was investigated. Using monoclonal antibody and immunoenzyme staining elevated p-53 level could be demonstrated in nuclei of embryonal carcinoma (84%) and seminoma components (56%). Most of the choriocarcinoma cases showed positive staining. Teratomas expressed this antigen negatively or scarcely. In seminomas the highest level of p-53 was stated in stage I. In contrast the opposite tendency could be demonstrated in embryonal carcinomas where p-53 was ++ positive in stage III. Between the high level of p-53 and clinical resistance a converse correlation could be stated because the resistant tumours expressed no or low, the sensitive tumours high level of p-53 protein (P 0.01). These results suggest that elevated p-53 expression could be a prognostic marker of sensitivity in testis cancer. PMID- 8721871 TI - [Fibrinolysis studied by an in vitro clot lysis test in polycythemia vera]. AB - Results obtained by a simple in vitro clot lysis test in polycythemia vera, polyglobulia, essential thrombocythemia and chronic immun thrombocytopenic purpura are reported. Increased lysis resistance of the platelet-rich clots was demonstrated in polycythemia and essential thrombocythemia, ostensibly caused by the high plasma level of plasminogen activator inhibitor-1. Following retraction of platelet-rich clots no further increase of lysis-resistance occurred in polycythemia and essential thrombocythemia. This phenomenon is most likely due to abnormal platelet function. The well known thromboembolic complications may at least partly result from the impaired fibrinolysis in diseases with high platelet counts. PMID- 8721870 TI - [Relationship between intestinal permeability and antibodies against food antigens in IgA nephropathy]. AB - Elevated antibody titers to food antigens and increased intestinal permeability in IgA nephropathy (IgA NP) suggest that these factors may be interdependent and play a role in the pathogenesis of the disease. In 1989 IgA and IgG antibodies against eight common food antigens were examined in the sera of 35 IgA NP patients and 12 controls using the ELISA method. Intestinal permeability was determined in 29 IgA NP patients and 20 controls using the 51Cr-EDTA test. Both examinations were carried out in 17 patients. Twenty-one patients allowed the intestinal permeability examinations to be repeated five years later. Compared with controls, significantly elevated IgA antibody titers against gliadin, soy, HAV (salt extracted antigens of oat flour) and ovalbumin were found. Intestinal permeability was significantly higher in IgA NP patients both in 1989 (3.86% +/- 0.29 vs. control, 2.72% +/- 0.23; p < 0.005) and in 1994 (4.57% +/- 0.63; p < 0.02) than in controls. A strong correlation was observed between the intestinal permeability and the IgA type of antibody titers against soy (p < 0.01; r = 0.72) and HAV (p < 0.02; r = 0.57). In patients without azotemia (serum creatinine > 150 mumol/l; n = 22) the decrease in creatinine clearance was significant only in the cases of increased intestinal permeability. On the basis of these data we assume that there is a connection between intestinal permeability and the production of IgA type antibody against food antigens in least a proportion of patients with IgA NP, and as a results of the lasting increase in intestinal permeability the production and the mesangial deposition of IgA immunocomplexes also increase. PMID- 8721872 TI - [Phototherapeutic use of excimer laser]. AB - The photorefractive excimer laser is not only able to alter the refractive power of the corneas but also suitable to treat superficial corneal pathologies. Authors performed phototherapeutic keratectomies (PTK) in 80 eyes of 77 patients. The age was on average (+/- S.D.) 49.4 +/- 22.03 years. The follow-up time varied between 3-24 months. All treatments were performed with the Aesculap Meditec MEL 60 ArF excimer laser, working in the 193 nm extreme ultraviolet-C range. Authors conclude that the best postoperative results can be achieved with PTK in cases of recurrent corneal erosions and superficial corneal opacities. Favourable outcome could be expected in treatment of infectious corneal ulcers. In cases of bullous keratopathy the subjective complaints decreased and it was successful to postpone the time of perforating keratoplasty. The PTK is not recommended in case of corneal Bowen disease and recurrent pterygia. A hyperopic shifts was observed in 2 eyes. Besides, the traditional surgical methods the phototherapeutic keratectomy is an important progress in treatment of superficial corneal pathologies. Applying the method, the recurrent erosions healed quicker, in most of the cases the transparency of the cornea could be reestablished in the pupillary area and the need for perforating keratoplasty could be postponed or avoided. PMID- 8721873 TI - [Loss of consciousness of unusual etiology in a young patient]. AB - The authors have presented the case of a young woman, who was observed because of repetitive loss of consciousness. The unbalance of the vegetative nervous system (parasympathetic overactivity) was responsible for the syncopic events, which could be prevented by the implantation of pacemaker successfully. The reoccurrence of loss of conciousness after 10 years could be attributed partly to the presence of dysfunction of pacemaker (undersensing+inhibition of myopotential), partly to a passing off the stress situation, with previous status of fever, and the consequent orthostatic hypotension caused by actually inadequat use of nitratvasodilator. PMID- 8721874 TI - [Albert Szent-Gyorgyi and flavonoid research]. PMID- 8721876 TI - [To the question of routine preoperative x-ray examination]. PMID- 8721875 TI - [Remembering Jozsef Balo (centenary of his birth)]. PMID- 8721877 TI - [Bronchiolitis obliterans organizing pneumonia]. PMID- 8721878 TI - [Preconceptional screening of sexually transmittable diseases]. PMID- 8721879 TI - [Prevention of anencephaly-spina bifida]. PMID- 8721880 TI - Umbilical cord osteocalcin in normal pregnancies and pregnancies complicated by fetal growth retardation or diabetes mellitus. AB - A homologous radioimmunoassay for human osteocalcin was used to measure cord serum osteocalcin concentrations in normal pregnancies and in pregnancies complicated by intrauterine growth retardation (IUGR) and diabetes. The mean osteocalcin concentrations in term newborns (100-110 micrograms/l) were comparable with levels that we previously measured in pubertal children. There was a small increase in mean osteocalcin concentrations during the third trimester of fetal life, with the maximum at 35 weeks; between weeks 36 and 41, the osteocalcin levels dropped by 13%. Osteocalcin was 20% lower in IUGR neonates than in age- or weight-matched newborns. The newborns of diabetic mothers had markedly lower osteocalcin concentrations than the weight-matched neonates, and 16 of the 19 samples were more than 1 SD below the mean of the gestational-age osteocalcin regression curve. Cord serum osteocalcin appears to be a useful parameter in studying normal and abnormal fetal mineralization. PMID- 8721881 TI - Maternal obesity and body composition of the neonate. AB - Obese women generally deliver heavier infants, but the body composition of these infants is unknown. The principal objective of this study was to determine if neonates of obese women have more adipose tissue. At 35-36 weeks of gestation, a fasting blood sample was collected from 37 pregnant women. Shortly after birth, the body fat of the neonates was measured with an infant total-body electrical conductivity (TOBEC) instrument using a prediction equation derived from 10 miniature pigs. At 6 weeks post partum, the infant body fat was measured a second time, and the body fat of each mother was measured using an adult TOBEC instrument. We found no differences between the obese (n = 16) and lean subjects (n = 21) in the concentrations of glycerol, beta-hydroxybutyrate, triglyceride, total cholesterol, high-density lipoprotein cholesterol, or glucose in the blood. However, the insulin concentration was elevated in the obese women (199 +/- 57 pmol/l) as compared with the lean women (128 +/- 68 pmol/l, p < 0.01). At birth, maternal adiposity (% body fat) was significantly associated with infant adiposity (r = 0.37, p < 0.05). However, by 6 weeks post partum the association no longer existed. Multiple regression analysis showed that maternal adiposity, fasting glucose level, and gestational age are independently associated at birth with infant adiposity. PMID- 8721882 TI - Brain and respiratory cilia: the effect of temperature. AB - We have used a modified photodiode method to measure ciliary beat frequency of human neonatal cilia, and have developed techniques to allow similar measurements of rat brain cilial function. The effect of temperature change, within the physiological range, on neonatal human respiratory cilia and neonatal rat brain cilia was studied. The mean (SD) ciliary beat frequency of neonatal respiratory cilia at body temperature was 14 (1.1) Hz and that of rat brain cilia 18.9 (1.3) Hz. Respiratory cilial beat frequency decreased by 9.1% and brain cilia by 13% on cooling to 32 degrees C compared to normal body temperature. Beat frequency increased by 8.5% in respiratory samples and by 12% in brain samples on warming to 40 degrees C compared to the frequency at normal body temperature. The Q10 of human neonatal respiratory cilia was 1.2 and that of rat brain cilia 1.3. PMID- 8721883 TI - Predicting the need for phototherapy in healthy mature neonates using transcutaneous bilirubinometry on the first postnatal day. AB - Since newborn infants are frequently sent home within 24-48 h after birth, the prediction of severe hyperbilirubinaemia, requiring treatment, is important. We tested the prediction of the need for phototherapy by measuring the yellow skin colour and the progression in yellow skin colour over 6 h using transcutaneous bilirubinometry (TcB), during the first 33 h of age. The first TcB reading at median 21 h (11-33 h, 5th and 95th percentiles) of age and the difference in readings over a 6-hour interval were reliable predictors of later phototherapy. An even better prediction was possible using the first TcB reading and the 6-hour difference in TcB readings. PMID- 8721884 TI - Hemodynamic adaptations at birth and neonates delivered vaginally and by Cesarean section. AB - In order to assess the possible influence of differences in delivery mode on cardiovascular adaptation at birth, we measured left ventricular output and its regional distribution in the major organs sequentially using an echographic technique during the first 96 h of life. We studied 27 normal newborns, of whom 15 were delivered vaginally and 12 by cesarean section. We also measured umbilical arterial and venous catecholamine concentrations. The umbilical arterial epinephrine and norepinephrine concentrations in the infants delivered vaginally were significantly greater than those in the infants delivered by cesarean section (epinephrine 1,195 +/- 208 vs. 565 +/- 81 pg/ml, p < 0.05; norepinephrine 11,832 +/- 3,819 vs. 5,153 +/- 1,400 pg/ml, p < 0.05). The left ventricular output and its regional distribution showed a similar pattern in the two groups, and there were no significant differences between them. These results indicate that the capacity of infants delivered by cesarean section to tolerate cardiovascular changes during the early neonatal period is comparable to that in infants delivered vaginally, even though there are significant differences in the catecholamine surge between these groups. PMID- 8721885 TI - Intrauterine growth retardation due to growth hormone deficiency in rats. AB - The effect of growth hormone (GH) on fetal growth was examined in the spontaneous dwarf rat (SDR) with the isolated GH deficiency. A 328-bp GH gene fragment containing the site of SDR mutation was amplified from fetal liver genomic DNA using PCR and digested with restriction enzyme SauI to distinguish the fetal genotype. When heterozygote (dr/+) males and females were crossed, a significant reduction in body weight of dr/dr fetuses was observed on days 20 and 21 (92 and 89% that of +/+ fetuses, respectively). Body weight reduction was also noticed on day 21 in dr/dr fetuses (80%) in comparison with that of dr/+ fetuses crossed from dr/dr females and dr/+ males. Unexpectedly, the body weight of dr/+ fetuses was found to be significantly less than that of +/+ fetuses on days 19 (94%) and 20 (97%), which reached the level of +/+ fetuses by day 21 of gestation. The fetal tail length was not affected by the GH status. These results provide direct evidence for the involvement of pituitary GH in the regulation of fetal body weight during late gestation, but not in fetal skeletal growth. PMID- 8721886 TI - Brain cell membrane Na+,K(+)-ATPase activity after inhibition of cerebral nitric oxide synthase by intravenous NG-nitro-L-arginine in newborn piglets. AB - This study investigated the effects of in vivo inhibition of cerebral nitric oxide synthase by intravenous administration of NG-nitro-L-arginine (NNLA) on the cell membrane Na+,K(+)-ATPase activity in the cerebral cortex of newborn piglets. NNLA was administered intravenously to 22 piglets at doses of 5 mg/kg (n = 3), 25 (n = 3), 50 (n = 4), 75 (n = 4), and 100 mg/kg (n = 2). Control animals (n = 6) received normal saline only. 90 min after infusion the cerebrum was obtained. The cerebral nitric oxide synthase activity, determined by measuring the conversion of [3H]-L-arginine into [3H]-L-citrulline in the brain homogenate, decreased from 9.1 +/- 2.0 pmol/mg protein/min in controls to 1.7 +/- 0.6 pmol/mg protein/min after the administration of 75 and 100 mg/kg NNLA. The Na+,K(+)-ATPase activity was measured in the P2 fraction of cortical tissue homogenate. The Na+,K(+) ATPase activity was within the normal range (48.3 +/- 4.9 mumol/mg protein/h) up to 75 mg/kg of NNLA. At a dose of NNLA of 100 mg/kg, the Na+,K(+)-ATPase activity decreased to 31.5 +/- 0.7 mumol/mg protein/h (p < 0.05). Four animals developed hypoxemia and lactic acidosis. The results demonstrate that inhibition of the cerebral nitric oxide synthase activity in vivo in newborn piglets by intravenous administration of NNLA did not affect the cortical cell membrane Na+,K(+)-ATPase activity up to a dose of 75 mg/kg. Doses of 100 mg/kg decreased the Na+,K(+) ATPase activity, probably by inducing cerebral hypoxia-ischemia. PMID- 8721887 TI - 3rd European workshop on drug information Helsinki, Finland, June 14-16, 1995. PMID- 8721888 TI - A study of the clinical outcome of patients with profound mental retardation gradually withdrawn from chronic neuroleptic medication. AB - In a naturalistic study of 23 severely and profoundly mentally retarded adult male patients undergoing slow "diagnostic" neuroleptic taper, it was determined that at least 60% could eventually be managed without psychoactive medication. However, many of these demonstrated a remarkably long, but nonetheless transient, period of worsening. This suggests that behavioral deterioration during drug reduction trials does not always indicate a need for chronic neuroleptic maintenance since these behaviors may return to baseline without pharmacological intervention. On the other hand, 40% demonstrated persistent (> 2 years) behavioral worsening. Those individuals who demonstrated persistent deterioration had been generally well controlled on neuroleptics, were somewhat older, and were receiving a higher baseline dose. Most of these persistently worsened subjects currently require some type of psychoactive medication (although only two have been returned to neuroleptics). PMID- 8721889 TI - Routine screening of thyroid function in patients hospitalized for major depression or dysthymia? AB - Our objective was to examine the clinical practice of testing thyroid function in a mixed community and referral psychiatry inpatient unit. We evaluated (1) the frequency of ordering screening, (2) the type of test, and (3) the incidence of thyroid function abnormality among those tested. We reviewed thyroid function tests obtained on 277 consecutive first time adult admissions to the psychiatric wards at Walter Reed Army Medical Center, from 1992 through 1993, who met the DSM III R diagnostic criteria for major depression or dysthymia. Of the 277 subjects, 260 (94%) had thyroid function tests performed which included evaluation of thyroid stimulating hormone (TSH). Seventeen patients (6.5%) had a TSH outside the normal range. Of these, there were two cases (0.4%) suggestive of hyperthyroidism and no overt cases of hypothyroidism. Eight patients had subclinical hypothyroidism (elevated TSH, normal T4). Although screening thyroid tests are often routine for depressed inpatients, our data suggest that thyroid screening may add little to diagnostic evaluation. Overt thyroid disease is rare among depressed inpatients. PMID- 8721890 TI - Risk factors for clozapine discontinuation among 805 patients in the VA hospital system. AB - The goal of this study was to determine if demographic or clinical factors collected at baseline on patients treated with clozapine would increase the risk of having clozapine discontinued for (a) lack of response, (b) side effects, (c) noncompliance, (d) concomitant illness, or (e) death. The subjects were 805 patients treated with clozapine at 96 Department of Veterans Affairs Hospital System facilities. Multiple logistic regression was used to determine if any of the baseline variables predisposed patients to discontinuation from treatment. Factors which were studied include age, race, history of inadequate response to traditional neuroleptics, history of substance abuse, and DSM-III-R Axis I diagnosis. Of the 805 patients started on clozapine 167 (20.7%) were discontinued from treatment. The only significant variable in the logistic regression model was race. This study finds that African American patients are more likely to have clozapine discontinued than non-African American patients, and there is a trend for prior history of inadequate response to traditional neuroleptics to predict clozapine discontinuation. We found no effect of substance abuse or dependence, diagnosis, or age on outcome in the overall patient group. In a post hoc analysis the African American patients had a significantly lower baseline white blood count than the non-African American patients, which could have explained the difference in clozapine discontinuation. The findings of this study support further investigation into the causes of ethnic differences in treatment outcome with clozapine. PMID- 8721891 TI - Impulse control disorders and attention deficit disorder in pathological gamblers. AB - Little systematic research has been done on psychiatric comorbidity of pathological gambling, an impulse control disorder. This report describes the occurrence of attention deficit disorder and impulse control disorders in 40 pathological gamblers in treatment for gambling problems and 64 controls. Diagnoses were made by structured interviews which utilized operationalized diagnostic criteria. An impulse control disorder other than pathological gambling was noted in 35% of the pathological gamblers, compared to 3% of the controls (p < .001). Compulsive buying (p < .001) and compulsive sexual behavior (p < .05) were significantly higher in pathological gamblers than controls. A strong association was seen among pathological gambling, attention deficit, and other impulse control disorders. Attention deficit disorder was seen in 20% of the pathological gamblers. Rates of impulse control disorders did not differ by gender. Implications of these high rates of comorbidity are discussed. PMID- 8721892 TI - Acute treatment response in outpatients with panic disorder: high versus low depressive symptoms. AB - The authors studied 75 outpatients with DSM-III-R panic disorder who had participated in a clinical trial and had been randomly assigned to receive fluvoxamine, cognitive therapy, or placebo for an 8-week period. They compared a group with high levels of depressive symptoms and a group with low levels of depressive symptoms. At baseline, patients with high levels of depressive symptoms were more likely to have severe phobic avoidance and to have higher scores on measures of anxiety, hyochondriasis, and disability. An important finding was that depressive symptoms improved at a rate which paralleled improvement in panic and anxiety. Likewise, the presence of depressive symptoms did not interfere with treatment response in panic disorder. Clinical implications of the findings are discussed. PMID- 8721893 TI - Clinical implications of antidepressant drug effects on sexual function. AB - Sexual dysfunction in a patient being treated with antidepressant medications may be due to the underlying depression, a coexisting medical illness, disruption of interpersonal relationships, or it may be a side effect of the medication. Almost all antidepressants are associated with sexual side effects that go above and beyond any symptoms that can be explained by the disease process itself. The incidence of such sexual side effects can be as high as 92% for some antidepressants. Some of the newer antidepressants currently on the market seem to have a lower incidence of sexual dysfunction as a side effect. In view of the fairly common occurrence of these unwanted effects, and their potential contribution to noncompliance, careful selection of antidepressant medications is necessary. A variety of treatment options is available, including decreasing the dosage of medication to the lowest-effective level, adjunctive medications (such as cyproheptadine, bethanechol, yohimbine, and amantadine, as well as other antidepressants) to counteract the adverse sexual effects, or switching to another antidepressant. The treatment of antidepressant-induced sexual dysfunction requires a creative approach on the part of the treating psychiatrist, and must be individualized to the patient. PMID- 8721894 TI - Clozapine treatment of euphoric mania. AB - Four patients with treatment-resistant euphoric mania characterized by elevated mood, hyperactivity, and rapid speech, refractory to standard treatments and anticonvulsants, showed dramatic improvement in symptoms when treated with clozapine. All developed enhanced functioning and new insight into previous psychopathology. This degree of improvement suggests that clozapine can be considered in the treatment of not only schizophrenia, schizoaffective disorder, and dysphoric mania but also classic euphoric mania unresponsive to traditional treatments. PMID- 8721895 TI - Encephalopathy associated with rapid decrease of high levels of lithium. AB - A case is described in which a highly detailed time course of serum lithium levels and mental status observations was consistent with neurotoxicity from the rapid decrease in high lithium levels rather than from the high lithium levels alone. The highest lithium level observed was 4.89 mEq/L; the patient remained oriented until about 40 h after hospital admission and the serum lithium level fell below 1.65 mEq/L. An EEG taken during the 2-day period of gross disorientation showed focal sharp epileptiform waves and paroxysmal bursts of slowing without full seizure. In view of these and previous complementary observations, and a rationale of similarity between hyperlithemia and hyponatremia, the potential toxicities of dialysis and abrupt lithium dose discontinuation should be considered in patients who develop high lithium levels after taking regular doses. PMID- 8721896 TI - Risperidone: a once-daily dosage schedule. PMID- 8721897 TI - A phase II controlled study of a combination of the immune modulator, lentinan, with didanosine (ddI) in HIV patients with CD4 cells of 200-500/mm3. AB - This study was carried out to assess the safety and efficacy of a combination of lentinan, an immune modulator, and didanosine (ddI) in a controlled study in HIV positive patients with CD4 levels of 200-500 cells/mm3. Didanosine was administered to HIV patients at doses of 400 mg/day (po) for six weeks (bid), then 2 mg of lentinan i.v. was added per week for 24-80 weeks. A control group (20%) received ddI only. A total of 107 patients were enrolled at three sites, and 88 patients started the ddI/lentinan phase. The combination caused significant increases in CD4 levels up to 38 weeks, whereas ddI alone was significant at the 5% level at 14 weeks. Based on these data, lentinan qualifies as a participant in future multi-drug studies in HIV. PMID- 8721898 TI - Serum levels of interleukin-8 as a marker of disease activity in patients with chronic sarcoidosis. AB - We determined the serum concentration of interleukin-8 (IL-8) in patients with chronic sarcoidosis and assessed its value as a marker of disease activity. The serum concentrations of IL-8 and tumor necrosis factor-alpha (TNF-alpha) were determined by enzyme-linked immunosorbent assays. Serum angiotensin-converting enzyme (ACE) activity was measured by a colorimetric assay in 16 patients with chronic sarcoidosis. Thirteen of the 16 patients with chronic sarcoidosis showed a serum IL-8 concentration higher than the normal range. The mean (+/- S. E.) serum IL-8 concentration in patients with active sarcoidosis (n = 6) was 18.7 +/- 1.3 pg/mL and that in patients with inactive sarcoidosis (n = 10) was 8.0 +/- 1.1 pg/mL; the difference was statistically significant (p = 0.002). An overall positive correlation between serum IL-8 concentration and serum ACE activity was apparent; however, only four of the 16 patients with sarcoidosis had a serum ACE activity greater than the normal range. TNF-alpha was detected in the serum of only two patients. The serum concentration of IL-8 may prove a useful marker for monitoring disease activity in sarcoidosis, especially in the chronic stage. PMID- 8721899 TI - Risk factors for myocardial infarction in cancer patients. AB - The increasingly long survival of cancer patients raises the question whether there might be an association between myocardial infarction (MI) and different types of cancer, especially since these different diseases share some of the same risk factors, like smoking. We describe a retrospective study in which we examined the autopsied hearts of 1642 cancer patients to determine the incidence of MI in different types of cancer and then studied the records of 106 of these cancer patients with complicated of fatal MI to determine their risk factors. The incidence of MI among the 1642 cancer patients was 6.5%. It was significantly higher in patients with squamous cell type cancers of the lung, head and neck, and urothelial tract cancer as compared with other types of cancers. The major coronary artery disease (CAD) risk factors in these patients were smoking, hypertension and left ventricular hypertrophy. Before therapy begins, cancer patients, especially those with squamous-cell-type cancer, should be screened for CAD risk factors, such as smoking, hypertension and left ventricular hypertrophy, and treated accordingly. PMID- 8721900 TI - Histamine excess symptoms in basophilic crisis of chronic myelogenous leukemia. AB - We encountered a patient with chronic myelogenous leukemia in basophilic crisis accompanied with histamine excess symptoms including bronchial asthma and gastric ulcer. The concentrations of histamine and histidine decarboxylase in leukemic cells containing granules typical for basophils were similar to those in mature basophils. His histamine excess symptoms rapidly disappeared concomitant with the reduction of blast cells after chemotherapy. We speculate that his histamine excess symptoms were induced by the leukemic cells. PMID- 8721901 TI - Production of granulocyte colony stimulating factor, granulocyte macrophage colony stimulating factor, and tumor necrosis factor alpha during remission and infections in patients with acute leukemia. AB - We measured circulating serum levels of granulocyte colony stimulating factor (G CSF), granulocyte macrophage colony stimulating comparable to the levels of these factors in 12 children with acute febrile infections without malignancy or hematological disorders and 15 age matched healthy controls. There were significantly elevated levels of G-CSF, GM-CSF and TNF alpha, in 12 children with infections without leukemia, as compared with controls. Also in 18 leukemic children with infections serum G-CSF and TNF alpha levels were significantly higher than those in the leukemic children without infection and healthy controls, whereas no significant difference was noted in the GM-CSF levels in these groups. Although elevation in TNF alpha levels in response to infections were similar in the children with and without leukemia, in the G-CSF levels lower elevation was noted in the leukemic children with infections as compared to the children with infections without leukemia. Despite leukopenia enhanced the production of G-CSF, even in leukopenic children with leukemia and infections, serum G-CSF levels were still lower than those for the children with infections without leukemia. We concluded that, the production of G-CSF and GM-CSF as a response to infection was deficient in the patients with acute leukemia in remission, probably due to the maintenance and reinforcement chemotherapy. Therefore, the use of recombinant G-CSF may be recommended in the infections of these patients. PMID- 8721902 TI - Klippel-Trenaunay-type syndrome: an eponym for various expressions of the same entity. AB - Klippel-Trenaunay (KT) or as it is also called Klippel-Trenaunay-Weber (KTW) syndrome is a rare congenital phacomatosis of uncertain aetiology and variable expression. The classical syndrome is a triad of congenital mesodermal abnormalities characterized by cutaneous angiomatous nevus commonly called port wine stain, venous varicosities and hypertrophy of soft tissue and/or overgrowth of bone of one or more limbs. Clinically, a diversity of phenotypes with subjacent malformations may be encountered all having in common abnormalities of the mesoblastic sheets that include angioblastic, lymphoblastic and osteoblastic lineages. Each of them may give rise to malformations that may exist alone or in an unlimited diversity of associations. We present three widely different clinical expressions of this syndrome and we support the proposal to group all these malformations under the eponym of Klippel-Trenaunay-Type (KTT) syndrome. PMID- 8721903 TI - Clinical characteristics of adult asthmatics requiring intubation. AB - Fifty eight admissions for 52 adult asthmatics who required intubation were reviewed for the years of 1988-1995 to examine factors related to specific clinical patterns and profile the course of these patients. Of the 56 admissions where patients were intubated for respiratory failure and/or cyanosis, 5 were associated with significant complications of mechanical ventilation/intubation (pneumothorax, subcutaneous emphysema, aspiration pneumonia, and laryngeal edema) and there were no fatalities. Patients > or = 35 years of age had significantly more profound respiratory acidosis in initial arterial blood gases (pH = 7.14 versus 7.23, p = 0.03). In contrast, patients with a history of drug abuse or psychiatric disorders had lower mean pCO2 (p < = 0.01). The overall mean length of intubation was 17.6 hours, while the overall mean hospital stay was 6.6 days. Longer intubation times were associated with the occurrence of major complications, female gender, and hospital administration of ipratropium. Hospital stay was correlated with length of intubation, later month of admission, and earlier year of admission. Common precipitating factors noted for first admissions were upper respiratory infections (61%), followed by allergy or smoke exposure (13%), compliance related problems (12%), and drug abuse/inhalation (6%). Inhaled anti-inflammatory drugs, oral steroids or either were taken at the time of admission by 35%, 35% and 65% of the patients, respectively. There was an even distribution of patients with respect to medical insurance coverage type, admissions per season or year, ethnicity, marital status, and gender. We conclude that severe asthma resulting in respiratory failure is common despite the frequent use of anti-inflammatory asthma medications. Mechanical ventilatory support can be administered safely in the majority of these patients and should be considered early in acute asthma. PMID- 8721904 TI - Correlation between steady-state plasma concentration of antituberculous drugs and age, inclusion of rifampicin in the treatment regimen, adverse drug reactions and other clinical parameters. AB - A regional hospital in Hong Kong examined the correlation between plasma concentrations of rifampicin, pyrazinamide, isoniazid and its metabolite hydrazine and age, inclusion of rifampicin in the treatment regimen, adverse drug reactions and other clinical parameters. One hundred eighty subjects with tuberculosis were admitted consecutively to the medical wards of the Prince of Wales Hospital over a one and a half year period. Elderly patients > 65 years were randomized into one of two treatments with and without rifampicin in addition to isoniazid, pyrazinamide and ethambutol; younger patients received all four drugs. Plasma antituberculous drug concentrations were determined using high performance liquid chromatography. Elderly patients taking rifampicin had a higher mean steady-state concentration of isoniazid, together with a higher incidence of adverse effects compared with those not taking rifampicin. No age related differences were observed for the other drugs. For the whole group, higher mean concentrations of hydrazine, rifampicin and pyrazinamide were associated with a higher incidence of adverse effects and the presence of coexisting diseases. It is concluded that in sick elderly patients with coexisting diseases, use of rifampicin in the antituberculous regimen should be accompanied by close monitoring for side effects, and that there may be an indication for use of lower dosages of antituberculous drugs in such patients. PMID- 8721905 TI - Hepatocyte growth factor and acute phase proteins in patients with chronic liver diseases. AB - Serum hepatocyte growth factor (HGF) levels are increased in patients with liver diseases. HGF has been recently reported to stimulate production of acute phase proteins such as alpha 2-macroglobulin and albumin of hepatocytes in primary culture. To clarify whether serum HGF concentrations have any relation to concentrations of acute phase proteins, we measured serum HGF and acute phase proteins in chronic liver diseases where the synthesis of many plasma proteins is decreased with the decline of liver function. Eighty three patients with chronic liver diseases and 20 normal individuals were examined for serum HGF, albumin, C reactive protein (CRP), alpha-fetoprotein (AFP), alpha 1-acid glycoprotein (alpha 1-AG) and alpha 2-macroglobulin (alpha 2-MG). Mean values for serum HGF in chronic hepatitis (CH), liver cirrhosis (LC) and hepatocellular carcinoma (HCC) were 0.37, 0.79 and 0.66 ng/mL, which were significantly higher than those in controls (p < 0.05, p < 0.0001 and p < 0.0001, respectively). The levels of CRP increased in parallel with the progression of chronic liver diseases. Levels of alpha 2-MG were not changed in patients with CH Or LC, while those in patients with HCC were significantly higher than in controls or LC (p < 0.01 and p < 0.05, respectively). Serum HGF showed a positive correlation with CRP and a negative correlation with albumin. However, no relations between HGF and alpha 2-MG were observed. These data suggest that serum levels of acute phase proteins such as albumin and alpha 2-MG are more closely associated with the degree of hepatic dysfunction than serum HGF levels. PMID- 8721906 TI - Experimental autoimmune hepatitis in mice following immunization with syngeneic liver proteins. AB - In this study, we describe the successful induction of experimental autoimmune hepatitis by immunizing inbred strains of mice with syngeneic liver antigens. Hepatic lesions, presumably based on autoimmune mechanisms, induced after 3 weekly immunizations were of only moderate degree. Of the various mouse strains used, C57BL/6 mice showed the most prominent histological changes of the liver, which were accompanied by elevated serum alanine aminotransferase levels. This system offers a potential model for further investigating experimental autoimmune hepatitis. PMID- 8721907 TI - Advances in computer technology: impact on the practice of medicine. AB - Advances in computer technology provide a wide range of applications which are revolutionizing the practice of medicine. The development of new software for the office creates a web of communication among physicians, staff members, health care facilities and associated agencies. This provides the physician with the prospect of a paperless office. At the other end of the spectrum, the development of 3D work stations and software based on computational chemistry permits visualization of protein molecules involved in disease. Computer assisted molecular modeling has been used to construct working 3D models of lens alpha crystallin. The 3D structure of alpha-crystallin is basic to our understanding of the molecular mechanisms involved in lens fiber cell maturation, stabilization of the inner nuclear region, the maintenance of lens transparency and cataractogenesis. The major component of the high molecular weight aggregates that occur during cataractogenesis is alpha-crystallin subunits. Subunits of alpha-crystallin occur in other tissues of the body. In the central nervous system accumulation of these subunits in the form of dense inclusion bodies occurs in pathological conditions such as Alzheimer's disease, Huntington's disease, multiple sclerosis and toxoplasmosis (Iwaki, Wisniewski et al., 1992), as well as neoplasms of astrocyte origin (Iwaki, Iwaki, et al., 1991). Also cardiac ischemia is associated with an increased alpha B synthesis (Chiesi, Longoni et al., 1990). On a more global level, the molecular structure of alpha crystallin may provide information pertaining to the function of small heat shock proteins, hsp, in maintaining cell stability under the stress of disease. PMID- 8721908 TI - Primary brain malignant lymphoma newly diagnosed during pregnancy. AB - Clinical brain tumors become symptomatic or show acceleration of symptoms in pregnant women less often when compared with those in non-pregnant women. We describe the clinical findings and MR imaging of a rare case of malignant lymphoma arising in the brain of a pregnant woman. PMID- 8721909 TI - Ultrasound-detected polycystic ovarian syndrome associated with depleted basal level but enhanced response of gonadotropin. AB - A patient with ultrasound-detected polycystic ovaries and anovulatory oligomenorrhea had extremely low luteinizing hormone (LH) levels, but a heightened sensitivity to gonadotropin-releasing hormone (GnRH). Of interest is the finding of undetectable basal LH levels. This rare presentation may suggest that polycystine ovarian syndrome (PCOS) features may be a consequence of the increased response of LH, independently on basal levels of LH. PMID- 8721910 TI - Recurrent abortion associated with 1qh+. PMID- 8721911 TI - [Intravascular therapies]. PMID- 8721912 TI - [Dietetics and neurology]. PMID- 8721913 TI - [Neurological complications associated with vitamin B1 deficiency (thiamine)]. PMID- 8721914 TI - [Epilepsy and dementia. An atavistic association]. AB - We carried out a brief historical review as to the meaning of the association between epilepsy and dementia and of 'epileptic dementia'. We also reviewed the literature published in the last decade on this theme, analyzing whether chronic recurrence of epileptic fits brings about cognitive deterioration. We concluded by affirming that 'epileptic dementia' was a mixed bag containing numerous diseases and syndromes with symptoms of both. Only exceptionally could epileptic fits bring on cognitive deterioration and this usually only slight. A great many factors and not just recurrent epileptic fits determine cognitive prognosis in epileptics. PMID- 8721915 TI - [Drug utilization studies]. AB - Studies in drug utilization allow us to study different aspects of the use of drugs and to implement ways of improving therapeutic quality, bearing in mind the fact that health resources are limited. The aim of this work is to give a panoramic view of the various aspects of medicine use studies: origins, aims, methodology. PMID- 8721916 TI - [Diet and neurology (I). Basis for nutrition]. PMID- 8721917 TI - [Survey of morbid obesity: current knowledge]. PMID- 8721918 TI - [Optimizing a diagnostic test using the receiver operating characteristic analysis and information theory]. PMID- 8721919 TI - [Bibliometric evaluation of the Spanish scientific output in neurosciences. Analysis of the publication with international readership between 1984 and 1993]. AB - A bibliometric study was made of the Spanish scientific output in the Neurosciences between 1984 and 1993. The material studied was selected in accordance with the Science Citation Index (SCI) for the year 1993 (Neurosciences section) and from the journals considered to be a "priority' by the data base Excepta Medica CD: Neurosciences. The usual bibliometric rules were used: price's law of scientific literature growth, Bradford's law of scientific literature dispersal and Lotka's law of author productivity. The national participation indices (IPa) in the leading publications with an international readership, the % SUP index (% of documents included in 10% of the world top quality production), the authors/articles index, institutional contributions and the geographical origin of these were all analyzed also. Our results showed an exponential growth in the Spanish neuroscientific literature (correlation coefficient: 0.981) and increasing IPa (period 1984-1988; 0.88; period 1989-1993: 1.59). A total of 1,488 original documents were analyzed. There was an impact factor (FI) of over 3 in 20.8% of the articles, with an average FI of 2.701. The % SUP was 6.07. The journal with the largest numbers of original papers in Brain Research (Bradford's first zone) with 186 articles and the largest IPa was in the Journal of Pineal Research (8.403). The total number of authors was 2,470, total signatures of authors were 5,958 (3.04% of the authors had a productivity index (IP) > or = 1 and 57% an IP = 0. The average number of authors/article was 4.02. The origin of the articles was: University (56.77%), Hospital (26.73%) and CSIC (14.95%). Scientific production was concentrated in the CCAA of Madrid and Cataluna 55.93%, followed by Andalucia, Castilla-Leon and Valencia (22.57%). PMID- 8721920 TI - [Latent periods in attending patients with acute cerebrovascular disease. A multicenter study]. AB - Treatment of stroke is effective when given straight away at the onset of symptoms. Our objective was to find what the present latent periods in the medical attention of stroke patients are. A study was made in 18 hospital neurology units. The time elapsed between onset of the stroke and being attended by a doctor in the Hospital Emergency Department, by a neurologist and in being admitted to hospital was determined. 794 patients were studied. The average time lapse before attention by the first doctor was 10.7 (0.1-504) hours, in receiving attention in the Emergency Department 14.1 (0.1-505), in being attended by a neurologist 24.3 (0.2-510) hours and in being admitted to hospital 21.3 (0-511) hours. In the first six hours 68.7% of the patients were seen by the first doctor, 58.2% went to the Emergency Department, 22.7% were attended by a neurologist and 22.3% were admitted to a Neurology Unit. The average time between arrival at the Emergency Department and seeing a neurologist was 10.5 (0-67.5) hours. To improve the treatment of stroke, reduction of the latent periods is necessary. PMID- 8721921 TI - [Latent period before hospital admission in cerebrovascular pathology: determining factors]. AB - OBJECTIVE: Establish the factors determining the latent period before hospital admission of patients with cerebrovascular disorders. MATERIAL AND METHOD: One hundred patients with cerebrovascular disorders admitted to the Hospital de la Santa Creu i Sant Pau, Barcelona were analyzed. All patients were systematically interviewed, evaluating: age, sex, place of residence, initial condition, type of transport to hospital, vascular risk factors, past history of cerebrovascular disorders, time of onset of symptoms, sleeping-wakefulness state, initial symptom, accompanying symptoms, initial course of illness, complications whilst in hospital, clinical condition and functional ability on leaving hospital, and duration of stay in hospital. All these factors were compared according to the latent period before admission (6 hours or less, or more than 6 hours). RESULTS: Patients with a latent period before admission of 6 hours or less more frequently had: changes in the level of consciousness (p = 0.04), motor disorders (p < 0.01), stable course (p = 0.04), systemic complications (p = 0.02) and were severely affected (p > 0.01). Patients with a latent period of more than 6 hours before admission more frequently: were smokers (p > 0.01), had a clinical course in stages (p < 0.01) and no limitation (p = 0.02). The stay in hospital lasted longer in the first group of patients. No differences were found when comparing: age, sex, place of residence, state at onset, type of transport to the hospital, vascular risk factors apart from smoking, past history of cerebrovascular illness, time of onset of symptoms, state of sleeping-wakefulness, initial symptoms and accompanying symptoms apart from changes in level of consciousness and motor defect. CONCLUSIONS: Factors determining delay in the hospital admission of patients with cerebrovascular disorders are: smoking, the clinical picture (alteration of consciousness, motor disorders) and the initial course of symptoms. PMID- 8721922 TI - [Some epidemiological aspects of myasthenia gravis in Cuba]. AB - We estimated the rates of prevalence, incidence, mortality and lethality for myasthenia gravis in Cuba based upon previous epidemiological studies carried out in eight of Cuba's fourteen provinces: Pinar del Rio, Havana City, Havana, Matanzas, Cienfuegos, Sancti Spiritus, Camaguey and Guantanamo. Rates of incidence, prevalence and mortality were 4.52, 29.22 and 0.72 per million inhabitants respectively. Lethality rate was 10.77%. PMID- 8721923 TI - [Daily defined doses of parkinsonian drugs in Alcoi]. AB - BACKGROUND: Some authors have suggested that the use of DDD (defined daily doses) of L-dopa may be useful as an indicator of the frequency of Parkinson's disease. AIM: To determine the measurement for L-dopa and selegiline in Alcoi. MATERIAL AND METHODS: We have obtained the intakes for L-dopa and selegiline between January and December, 1990 and we have calculated the DDD/1,000 inhab/day. RESULTS: During the study period, we have found that the mean intakes were 18,000.969 mg and 53,800 mg for L-dopa and selegiline, respectively. And so, the L-dopa intake in Alcoi was 0.60 DDD/ 1,000 inh, and the selegiline intake was 0.21 DDD/1,000 inh. DISCUSSION: Comparing the other data of the literature, specially in scandinavian countries, the obtained intake for L-dopa was low, and that could indicate that the frequency of the disease in Alcoi was lower. PMID- 8721924 TI - [Cerebrovascular accident in young adults. A study of 52 cases]. AB - OBJECTIVE: A retrospective study of ischaemic and haemorrhagic cerebral vascular accidents in patients under 45 years of age, admitted to hospital Severo Ochoa (Area sanitaria 9 de la Comunidad de Madrid) during a 6 year period. METHOD: 23 CVAs in women and 29 in men were studied. The aetiology of each episode was determined following preestablished criteria. RESULTS: Aetiology: cardiac 13 (25%), atherosclerosis: 8 (15%), haematological: 6 (11%), non-atherosclerotic: 8 (15%), migraine: 1 (2%), hypertensive: 2 (3%), unknown: 15 (29%). There was a predominance of men in the patients with atherosclerosis. These also had more cardiovascular risk factors and were older than the other patients. All patients with CVA due to antiphospholipid antibodies had multiple CVAs. Examinations used to establish the aetiology were: echocardiogram, angiography, carotid doppler, hypercoagulability studies and the clinical history. CONCLUSIONS: In spite of extensive study, 30% of the CVAs in young people still have no aetiological diagnosis. In general in ischaemic CVAs, there is good recovery of the neurological defects and a low mortality. The detection and oral anticoagulation of patients with a source of cardiac emboli, and the elimination of cardiovascular risk factors would probably considerably reduce the incidence of ischaemic stroke in young adults. PMID- 8721925 TI - [Pseudotumor cerebri secondary to cerebral venous defects not identified by magnetic resonance]. AB - Pseudotumour cerebri is the name of a syndrome characterized by headache and papilloedema, with normal cerebral CT/MR studies and CSF with a high pressure and normal laboratory findings. We describe four patients who fulfilled the diagnostic criteria of this condition (including normal 0.5T MR studies). They all had cerebral angiograms showing minor abnormalities localized to the level of the superior longitudinal sinus. All improved on treatment with anticoagulants and steroids. In view of these findings we consider that in cases of pseudotumour cerebri without a clear aetiological factor, an angio MR study should be done, or if this technique is not available, a cerebral angiogram should be done, to exclude cerebral venous drainage defects. PMID- 8721926 TI - [Low flow cavernous sinus fistula. Treatment by highly selective embolization]. AB - Carotid-cavernous sinus fistulae are anomalous communications between the carotid artery and the cavernous sinus. They are classified according to whether they are traumatic or spontaneous, but distinction in relation to the anatomical origin of the branches of the carotid artery which supplies them is more important. The anatomy of the regional vasculature is analysed in this article. The vessels most frequently involved in spontaneous dural fistulae are the internal maxillary artery (from the external carotid) and the meningohypophyseal trunk (from the internal carotid) which anastomose respectively with the inferior cavernous sinus artery and with the ascending pharyngeal artery. We present two cases of spontaneous carotid-cavernous fistulae. Both were type D of Barrow's classification, and in view of progressive changes in the visual fields, it was decided to proceed with invasive treatment by highly selective embolization of the branches of the external carotid artery, which is currently the treatment of choice in such cases. Good results were obtained. PMID- 8721927 TI - [Neuropathic beriberi as a complication of surgery of morbid obesity]. AB - We describe a women of 22 years of age who had had a vertical gastroplasty (as treatment for morbid obesity). She was admitted to hospital with a 4 week history of nausea and vomiting of food. Treatment with intravenous dextrose, without vitamin supplements was started. One week later she complained of diplopia, paresthesia and weakness of the limbs. All investigations proved to be normal. A deficiency state was suspected, probably Wernicke's encephalopathy, although no alterations were seen in her mental state. We started treatment with high doses of parenteral thiamine, other vitamins and a suitable diet. The treatment was followed by complete recovery. Few neurological complications have been described in association with vertical gastroplasty. The commonest are polyneuropathies. The probable deficiency origin of these is considered. We emphasise the importance of vitamin supplements following the surgical treatment of morbid obesity to avoid the development of deficiency states. Wernicke's encephalopathy is due to thiamine deficiency. It may be associated with any type of malnutrition, not only with chronic alcoholism. The full clinical triad which is diagnostic of this condition is only present in one third of the cases. When the condition is suspected on clinical grounds treatment should be started early to avoid the occurrence of irreversible secuelae. PMID- 8721928 TI - [Contralateral hyperhidrosis secondary to the pontine infarct]. AB - Unilateral hyperhydrosis due to a cerebral infarct is a clinical sign rarely described in the literature, the significance of which is unknown and may be due to a lesion of the crossed sympathetic inhibitory tract. We describe the case of a patient with contralateral hyperhydrosis due to a hemipontine infarct. PMID- 8721929 TI - [Cerebral infarct as a presentation of a hereditary defect of type II C protein]. AB - The case of a 50 year-old patient with a cerebral infarct related to a deficit in type II C protein is described. The patient showed no other vascular risk factor and the cardiological study, which included a transthoracic and a transoesophagic echogram ruled out the presence of embologenic cardiopathy. A family study detected the presence of a deficit of C protein in 6 of the 8 sons and in the patient's sister who had a deep vein thrombosis at the age of 54. A hereditary deficiency in C protein was confirmed. We consider it necessary to perform hypercoagulability studies which include the determination of C protein in patients under 55 years of age with cerebral infarcts of unknown cause, especially when there is a family history of thrombosis. PMID- 8721930 TI - [A 21-year-old man with myoclonic jerks cognitive deterioration and progressive ataxia]. PMID- 8721931 TI - [Hemichorea-hemiballism by hemorrhage in nucleo-thalamic syndrome. Letter]. PMID- 8721932 TI - Psychological stress and the onset of IDDM in children. AB - OBJECTIVE: The purpose of the study was to determine whether psychosocial stress during different life periods could be a risk factor in the etiology/pathogenesis of insulin-dependent diabetes mellitus (IDDM) in children. RESEARCH DESIGN AND METHODS: In a population-based sample of 67 case patients 0-14 years of age and 61 matched healthy control subjects, life events during the entire lifespan before the onset of IDDM were recorded as well as measures of child behavior before onset, social support, and family function. RESULTS: Negative life events occurring during the first 2 years of life, life events with difficult adaptation, child behavioral deviances, and a more chaotic family function were more common in the case group. A stepwise logistic regression indicated that negative life events in the first 2 years increased the risk of IDDM and that premorbid child behavior as well as dysfunctional hierarchical family pattern affect the risk. CONCLUSIONS: Stress early in life may increase the risk for IDDM, presumably by affecting the autoimmune process. To confirm these results, it is necessary to make a truly prospective study. PMID- 8721933 TI - Psychosocial status of children with diabetes in the first 2 years after diagnosis. AB - OBJECTIVE: The purpose of this study was to determine the pattern of adjustment over time of a cohort of children with newly diagnosed diabetes compared with a cohort of peer-selected children without diabetes over the first 2 years after the diagnosis of insulin-dependent diabetes mellitus (IDDM). RESEARCH DESIGN AND METHODS: Children (n = 89 with IDDM, n = 53 without IDDM) ages 8-14 years were studied with the Children's Depression Inventory, State-Trait Anxiety Inventory for Children, Child and Adolescent Adjustment Profile, Self-Perception Profile for Children, and a general health scale. Initial data were collected within 6 weeks of the diagnosis of IDDM and at 3, 6, 12, and 24 months thereafter. RESULTS: There were no demographic differences between the two groups. Initially, children with diabetes were more depressed, more dependent, and more withdrawn than their peers. By 1 year postdiagnosis, there were no significant differences in psychosocial status between the two groups. By 2 years postdiagnosis, depression, dependency, and withdrawal were significantly higher in children with diabetes than in their peers without IDDM. Self-perceived competence remained similar between the two groups at all time points. CONCLUSIONS: After an initial period of adjustment, children with IDDM have equivalent psychosocial status to children without IDDM, but by 2 years after diagnosis, they have experienced twice the amount of depression and adjustment problems as their peers. Interventions should be aimed at this critical period between 1 and 2 years postdiagnosis. PMID- 8721934 TI - Effect of aging on HbA1c in a working male Japanese population. AB - OBJECTIVE: To evaluate whether there is any change in HbA1c with age and to determine the effects of body mass index (BMI), exercise, and family history of diabetes on this change. RESEARCH DESIGN AND METHODS: A cross-sectional survey of 7,664 male Japanese workers aged 20-59 years was performed. All subjects received a physical examination that included measurement of HbA1c as an indicator of plasma glucose level. The subjects were classified according to their ages and BMIs, and any relationship with HbA1c levels was evaluated. Information on physical activity and family history of diabetes was obtained by a questionnaire. RESULTS: In all BMI groups, HbA1c increased with age. The greatest increase in HbA1c was observed in the 40- to 49-year-old age-group in subjects with a BMI < or = 26 kg/m2 and in the 30- to 39-year-old age-group in subjects with a BMI > 26. HbA1c in the subjects aged 20-29 years did not change with BMI. In contrast, HbA1c in subjects aged 30-59 years was significantly higher in those with a BMI > 26 when compared with those with BMI < or = 20. The age-dependent increase in HbA1c was greater in subjects with a positive rather than negative family history of diabetes. This age-dependent increase was not affected by active participation in sporting activities of mild to moderate intensity for 3 or more days per week. CONCLUSIONS: The age-dependent increase in HbA1c may be a consequence of the aging process itself. Furthermore, both BMI and a hereditary predisposition to diabetes, but not active participation in sports, affect this age-dependent increase in HbA1c. PMID- 8721935 TI - Probucol normalizes cholesteryl ester transfer in IDDM. AB - OBJECTIVE: Cholesteryl ester transfer (CET) is pathologically increased in insulin-dependent diabetes mellitus (IDDM), and the resulting enrichment of the apolipoprotein B-containing lipoproteins with cholesteryl ester (CE) is believed to increase their atherogenicity. Because we have shown previously that treatment with the lipid-modifying antioxidant probucol normalizes CET in nondiabetic patients with hypercholesterolemia, we sought to determine whether the same beneficial effects could be achieved in IDDM. RESEARCH DESIGN AND METHODS: CET was measured by both mass and isotopic assay in eight normolipidemic (triglyceride, 102; cholesterol, 192; high-density lipoprotein [HDL] cholesterol, 45 mg/dl) IDDM patients (fructosamine 495 +/- 146 mumol/l; normal 174-286) before and after 2 months of treatment with probucol (1.0 g/day). RESULTS: Before treatment, CET was accelerated abnormally (P < 0.001). As expected, probucol decreased plasma (-13%; P < 0.025) and HDL2 cholesterol levels (-52%; P < 0.025) and the concentration of lipoprotein A-I particles (P < 0.025). In conjunction with these changes, CET fell dramatically in all subjects (mass assay: -94%; isotopic assay: -22%, P < 0.001) with no change in the mass of cholesteryl ester transfer protein (CETP) (pretreatment 2.91 +/- 0.97 vs. posttreatment 3.21 +/- 1.03 micrograms/ml). Glycemic control, however, improved significantly (fructosamine 409 +/- 85 mumol/l, P < 0.025). CONCLUSIONS: Because it is believed that accelerated CET promotes the formation of apolipoprotein B-containing lipoproteins enriched with atherogenic CE, the capacity of probucol to reverse this functional abnormality without adversely affecting glycemic control suggests that it has a place in the therapy of IDDM. PMID- 8721936 TI - Empirical validation for a family-centered model of care. AB - OBJECTIVE: To examine whether family relations and family-life stress predict adherence behaviors and metabolic control in youths by testing a theoretically- and empirically-based model. RESEARCH DESIGN AND METHODS: HbA1c levels of 157 youths 12-20 years of age with insulin-dependent diabetes mellitus (IDDM) were evaluated, along with treatment adherence and psychosocial family data based on interviews and self-reports. A family-centered model for IDDM care was analyzed with the EQS structural modeling statistical program. An additional exploratory model including HbA1c levels 6 months earlier was also tested. RESULTS: Positive family relations (high family cohesion and low family conflict), especially during the first years of illness, indirectly related to good metabolic control through positive adherence behaviors. High levels of family-life stress related to worse metabolic control directly, and also indirectly via poor family relations, which in turn related to poor adherence to treatment. Longer illness duration related indirectly to metabolic control via adherence to treatment. Longer illness duration also related to maternal perceptions of higher family life stress. Older youths perceived more family-life stress as well. The older age of the youths related indirectly to poor metabolic control through its association with poor adherence behaviors and less cohesive and more conflictual family relations. Results were consistent when earlier levels of metabolic control were included in the model, except that the direct relationship between family-life stress and metabolic control became nonsignificant. CONCLUSIONS: The findings suggest that a family-centered approach to diabetes care that facilitates positive family functioning and leads to lower levels of family-life stress may be useful in promoting optimal health outcomes. PMID- 8721937 TI - Elevated serum total renin is insensitive in detecting incipient diabetic nephropathy. AB - OBJECTIVE: To assess the influence of incipient diabetic nephropathy on the levels of total renin in serum. RESEARCH DESIGN AND METHODS: Fifty-five adult patients with insulin-dependent diabetes mellitus (IDDM) were examined in a cross sectional study. The main outcome measures were serum total renin concentration and urinary albumin excretion rate. RESULTS: The total renin concentrations in serum were significantly (P < 0.05) higher in 12 patients with microalbuminuria than in 43 patients without albuminuria, but this difference was significant only in men. There was a significant but weak positive correlation between urinary albumin excretion rate and serum total renin concentration in all patients (r = 0.412, P < 0.05, n = 55), but the sensitivity of high serum concentrations of total renin in detecting incipient nephropathy was low (17%). In the study group, two of the three patients suffering from proliferative retinopathy had the highest total renin concentrations in serum. CONCLUSIONS: Microalbuminuric patients have higher mean serum total renin concentrations than normoalbuminuric patients, but because of low sensitivity, high total renin concentration cannot be used for screening incipient diabetic nephropathy. PMID- 8721939 TI - Psychological impact of islet cell antibody screening for IDDM on children, adults, and their family members. AB - OBJECTIVE: To describe the psychological impact of positive islet cell antibody (ICA) screening results in children and adults, as well as their parents and spouses. RESEARCH DESIGN AND METHODS: The psychological impact of ICA screening results was assessed subsequent to subjects' being informed of ICA-positive (ICA+) status and was re-evaluated 4 months later. Impact was measured using the state subscale of the State-Trait Anxiety Inventory (STAI) for adults or the State-Trait Anxiety Inventory for Children (STAIC), as well as structured interviews. A total of 34 ICA+ children, 34 ICA+ adults, 33 parents, and 25 spouses were evaluated. RESULTS: At initial notification of ICA+ status, clinically and statistically significant anxiety was observed in ICA+ children and adults and their family members (P < 0.001). Parents of ICA+ children were more anxious than spouses of ICA+ adults (P < 0.05). Child and parent anxiety were significantly correlated (P < 0.05); more-anxious children lived with more anxious parents. No significant association was found between ICA+ adults' initial anxiety and their spouses' anxiety. For ICA+ participants and their family members, anxiety dissipated to normal levels in 4 months (P < 0.02). ICA+ children were less likely than parents to believe they would ever develop insulin dependent diabetes mellitus (IDDM). Nevertheless, 52% of ICA+ children and 24% of ICA+ adults endorsed lifestyle or behavior changes as a result of their IGA+ status. Behavior change was associated with greater initial anxiety in both children and adults (P < 0.05 for both). CONCLUSIONS: These data suggest that notification of ICA+ status has both emotional and behavioral impact. Initial notification of ICA+ status is associated with considerable anxiety in both ICA+ individuals and their family members. In most cases, this initial anxiety appears to dissipate to normal levels over time. However, many ICA+ individuals report initiating lifestyle or health behavior changes in an effort to delay or prevent IDDM onset. PMID- 8721938 TI - Effects of insulin on cholesterol synthesis in type II diabetes patients. AB - OBJECTIVE: To evaluate the effects of intensive insulin therapy and subsequent optimized metabolic control on daily urinary mevalonic acid (MVA) excretion, an index of whole-body cholesterol synthesis, and the acute effects of insulin on plasma MVA concentrations in type II diabetes. RESEARCH DESIGN AND METHODS: Ten (five men and five postmenopausal women) nonobese, normolipidemic (total cholesterol < 6.2 mmol/l, triglycerides < 2.82 mmol/l), type II diabetic patients in poor metabolic control (HbA1c > 10%, fasting plasma glucose > 11 mmol/l) and receiving sulfonylurea treatment were selected. The 24-h urinary MVA excretion and plasma lipid values were determined before and after intensive insulin therapy. The acute effects of insulin on plasma MVA concentrations were also evaluated during a 3-h euglycemic hyperinsulinemic clamp study. RESULTS: Urinary MVA excretion rates (mumol/24h) were 1.82 +/- 0.21 in control subjects and 2.49 +/- 0.35 (P < 0.01 vs. control subjects) and 1.78 +/- 0.28 in patients before and after intensive insulin therapy, respectively. Total cholesterol, low-density lipoprotein (LDL) cholesterol, and triglycerides decreased by 9, 8, and 12%, respectively, after blood glucose optimization. Acute insulin infusion during the euglycemic clamp studies reduced mean plasma MVA concentrations at 120 and 180 min by 29 and 38%, respectively (P < 0.01 for both vs. baseline). CONCLUSIONS: Our study demonstrates that in nonobese, normolipidemic, type II diabetic patients under poor metabolic control, an increased cholesterol synthesis is normalized by insulin therapy. Hyperinsulinemia in the presence of euglycemia acutely decreases the circulating levels of MVA, the immediate product of hydroxymethylglutaryl-CoA reductase activity and an index of whole-body cholesterol synthesis. PMID- 8721940 TI - Ginseng therapy in non-insulin-dependent diabetic patients. AB - OBJECTIVE: To investigate the effect of ginseng on newly diagnosed non-insulin dependent diabetes mellitus (NIDDM) patients. RESEARCH DESIGN AND METHODS: In this double-blind placebo-controlled study, 36 NIDDM patients were treated for 8 weeks with ginseng (100 or 200 mg) or placebo. Efficacy was evaluated with psychophysical tests and measurements of glucose balance, serum lipids, aminoterminalpropeptide (PIIINP) concentration, and body weight. RESULTS: Ginseng therapy elevated mood, improved psychophysical performance, and reduced fasting blood glucose (FBG) and body weight. The 200-mg dose of ginseng improved glycated hemoglobin, serum PIIINP, and physical activity. Placebo reduced body weight and altered the serum lipid profile but did not alter FBG. CONCLUSIONS: Ginseng may be a useful therapeutic adjunct in the management of NIDDM. PMID- 8721941 TI - Manufactured shoes in the prevention of diabetic foot ulcers. AB - OBJECTIVE: To evaluate the efficacy of manufactured shoes specially designed for diabetic patients (Podiabetes by Buratto Italy) to prevent relapses of foot ulcerations. RESEARCH DESIGN AND METHODS: A prospective multicenter randomized follow-up study of patients with previous foot ulcerations was conducted. Patients were alternatively assigned to wear either their own shoes (control group, C; n = 36) or therapeutic shoes (Podiabetes group, P; n = 33). The number of ulcer relapses was recorded during 1-year follow-up. RESULTS: Both C and P groups had similar risk factors for foot ulceration (i.e., previous foot ulceration, mean vibratory perception threshold > 25 mV). After 1 year, the foot ulcer relapses were significantly lower in P than in C (27.7 vs. 58.3%; P = 0.009; odds ratio 0.26 [0.2-1.54]). In a multiple regression analysis, the use of therapeutic shoes was negatively associated with foot ulcer relapses (coefficient of variation = -0.315; 95% confidence interval = -0.54 to -0.08; P = 0.009). CONCLUSIONS: The use of specially designed shoes is effective in preventing relapses in diabetic patients with previous ulceration. PMID- 8721942 TI - Nesidioblastosis causing reversal of insulin-dependent diabetes and development of hyperinsulinemic hypoglycemia. PMID- 8721943 TI - Well-child care and IDDM: whose responsibility? PMID- 8721944 TI - Preventing amputation in the patient with diabetes. PMID- 8721945 TI - Acute cataracts in newly diagnosed IDDM in five children and adolescents. PMID- 8721946 TI - Lessons for the American Diabetes Association from the health care debate. PMID- 8721947 TI - Elevation and fluctuation of serum CA 19-9 and SPAN-1 levels after administration of sulfonylurea in a diabetic patient with Lewis(a-,b-) blood phenotype. PMID- 8721948 TI - No increase of IDDM incidence among Swiss military conscripts. PMID- 8721949 TI - Absence of the Gly40-Ser mutation in the glucagon receptor among diabetic patients in the Netherlands. PMID- 8721950 TI - American Diabetes Association scientific sessions, 1995. PMID- 8721951 TI - Dimethyl sulfoxide-induced apoptosis in human leukemic U937 cells. AB - Dimethyl sulfoxide (DMSO), which induces differentiation of myeloid cells, was found to cause apoptosis in human leukemic U937 cells. Apoptosis was assessed by DNA electrophoresis and flow cytometry. The time needed to induce apoptosis varied from a few hours to 2-3 days, depending on the concentration of DMSO used. The plasma membrane remained intact long after DNA fragmentation had occurred. DMSO-induced apoptosis was inhibited by zinc ions and, to a lesser extent, by the protein kinase C activator: phorbol 12-myristate 13-acetate (PMA). Cycloheximide and actinomycin D did not prevent DMSO-induced apoptosis, showing that U937 cells do not require protein or RNA synthesis to undergo apoptosis. DMSO induced apoptosis despite the expression of the anti-apoptotic Bcl-2 protein in U937 cells. The amount of Bcl-2 remained unchanged during DMSO-induced apoptosis. PMID- 8721952 TI - 'Blind' multicenter evaluation of the prognostic value of DNA image cytometric and morphometric features in invasive breast cancer. AB - The prognostic value of classical prognosticators in 92 invasive breast cancers with long follow-up has been compared with a number of DNA-related image cytometric features (2c deviation index (2cDI)), Auer (histogram) type, 5c exceeding rate (5cER), DNA malignancy grade (MG), and the morphometrically assessed mitotic activity index (MAI) and mean nuclear area (MNA). The quantitative analyses were carried out without knowledge of the outcome nor of clinicopathological features. The cytometric analyses and assessments of MNA were performed on Feulgen stained, 4-microns thick tissue sections in Dresden. The MAI was assessed in Amsterdam. Thereafter, the survival data were matched with the classical variables, the cytometric features, the MAI and the MNA in univariate and multivariate analyses. Lymph node status (LN) and tumour stage were significantly different, tumour stage being the strongest discriminator between survivors and non-survivors (Mantel-Cox value = MC = 30.7, P < 0.0001). Grade was (just) significant (P = 0.05). The cytometric features, the MAI and the MNA were all highly significant prognosticators. The strongest cytometric feature was the Auer type followed by the 5cER, and the 2cDI. The MNA was also significant. The MAI was the strongest single prognostic factor in this blind analysis (MC = 50.8, P < 0.0001), and the MAI combined with the lymph node status, tumour stage and the cytometric features Auer type and 2cDI was the best multivariate combination (MC = 94.0). Separate analyses of the lymph node negative (LN-, n = 39) and positive (LN+, n = 53) groups showed that the 5cER and MAI were the most important in the LN- and the MAI in the LN+ patients. PMID- 8721953 TI - Cell proliferation and ploidy of human solid tumours: methodological experience with in vivo bromodeoxyuridine and DNA flow cytometry. AB - A sequential procedure for single and multiparameter flow cytometry (FCM) that allows detailed cell proliferation and DNA ploidy studies of human solid tumours is reported here. This description includes time collection and storage, tissue disaggregation and staining as well as FCM analysis of samples. The overall feasibility, together with some critical aspects of the DNA/bromodeoxyuridine (BrdU) in vivo assay for cell kinetic studies in human solid tumours, are reported, based on our experience in recent years. We found that the BrdU in vivo method, coupled with bivariate FCM for measurements, is a valuable approach for a 'dynamic' assessment of tumour cell proliferation in the clinical setting. Routine measurements can be achieved providing that well standardised tissue disaggregation and immunolabelling procedures are performed. In different solid tumours, cytokinetic results were satisfactory in 85% of cases as far as the BrdU labelling index is concerned while 70% were acceptable for DNA synthesis time and tumour potential doubling time. Causes of failure are also considered and discussed. In order to guarantee the finest detection of aneuploidy, routine high resolution single-parameter DNA analysis is suggested. Our experience with further improvements in cell kinetic studies based on the possibility of gating the BrdU/DNA analysis for a subpopulation of interest (i.e. cytokeratin-positive cells in epithelial tumours) is also reported. PMID- 8721954 TI - Endocervical columnar cell intraepithelial neoplasia (ECCIN). 3. Interobserver variability in feature use. AB - In our previous studies on endocervical columnar cell abnormalities, the ability of 5 cytotechnicians was tested to distinguish between cases of: no abnormalities, different grades of intraepithelial endocervical columnar cell atypias and invasive adenocarcinoma. On the basis of stepwise multiple regression analysis, nuclear chromatin distribution, variation in cellular and nuclear size, cytoplasmic eosinophilia and architectural features such as cell-crowding, cluster formation, formation of gland-like structures and pseudostratification appeared to be of primary diagnostic importance to discriminate between no abnormalities, different grades of cervical columnar cell atypias, adenocarcinoma in situ and adenocarcinoma. The present paper reports the results of a study which was designed to assess the individual performance in feature use and classification of endocervical lesions. The results of this study indicate that observers use different characteristics and different grading for the level of expression of these characteristics. However, all observers illustrated a strong relationship between the presence and the expression grade of a certain feature and the degree of endocervical columnar cell atypia. With acceptance of one grade of difference in grades of severity, an overall correct diagnosis was made in 87.6% of cases (range 80-98.2%). PMID- 8721955 TI - Immunocytometric determination of the alpha and beta isoforms of human DNA topoisomerase II: influence of different fixatives. AB - The immunoreactivity of the alpha and beta isoforms (170 and 180 kDa, respectively) of human DNA topoisomerase II (topo II) toward specific monoclonal antibodies (MoAbs), was investigated in HeLa cells. The extent of antigen preservation achieved with the dehydrating fixatives, ethanol and acetone, was compared with that obtained with the cross-linking agent paraformaldehyde. Binding of each MoAb to the relative isozyme was detected by indirect immunofluorescence labelling and quantified by flow cytometry. The amount of antigen detectable was calculated by normalizing the mean fluorescence intensity of samples incubated with specific MoAbs, to the respective background fluorescence. For both isozymes, acetone provided the highest immunofluorescence/background ratio. A strong reduction in the immunoreactivity of the 180-kDa isoform was observed after ethanol, and to a minor extent after paraformaldehyde fixation, while reactivity of the 170-kDa isoform was not significantly affected. Cell cycle distribution of each protein was assessed by dual-parameter analysis of immunofluorescence vs. DNA content. No evident difference in the cell cycle distribution of each isozyme was found with each fixation protocol. At the morphological level, the pattern of immunofluorescence showed that the localization of each isozyme was very similar for all these fixatives. These results confirm the great sensitivity of the 180-kDa isoform to degradation, and indicate that fixation has to be carefully chosen in order to determine the relative amount of topo II isoforms with immunocytometric techniques. PMID- 8721956 TI - Expression of c-erbB2 oncogene product in different tumours and its standardised evaluation. AB - In order to evaluate the reliability and reproducibility of the immunohistochemical demonstration of the c-erbB2 oncoprotein and define criteria for positivity assessment in paraffin-embedded material from different tumours, we examined immunohistochemically 79 gastric, 100 colorectal and 20 bladder carcinomas using 5 different c-erbB2 antibodies. To determine criteria for the evaluation of c-erbB2 positivity, the observed immunohistochemical staining was compared with the actual protein prevalence, determined by Western blot analysis of each tumour. The comparison of protein levels obtained from Western blot with corresponding immunohistochemistry revealed that positive staining can only be considered to be specific for c-erbB2 oncoprotein if: (i) there is a definite staining along the tumour cell membrane with no predominant immunoreactivity inside the tumour cell cytoplasm, (ii) the intensity of immunoreactivity is comparable with a positive standard (e.g. in this study breast carcinoma) and (iii) the surrounding normal, non-neoplastic tissue shows no immunoreactivity. The graduation of staining intensities did not correlate to the actual amount of protein detected in Western blots. This study stresses the importance of defined and controlled criteria for evaluation of c-erbB2 oncoprotein positivity in immunohistochemistry. PMID- 8721957 TI - Lower extremity angiography: improved image quality and outflow vessel detection with bilaterally antegrade selective digital subtraction angiography. A blinded prospective intraindividual comparison with aortic flush digital subtraction angiography. AB - RATIONALE AND OBJECTIVES: To introduce routine bilaterally antegrade selective stationary digital subtraction angiography (DSA), and prospectively compare it with unselective stationary DSA in the detection of calf arteries and assess additional time and complication rate. METHODS: Twenty-five patients received one unselective and two separate antegrade selective studies of each calf. Images were evaluated for image quality, number of depicted run-off vessels, and potential crural bypass recipient arteries. RESULTS: Bilaterally antegrade selective DSA was significantly superior in image quality and motion artifacts (P < 0.01). The number of adequately depicted run-off arteries per calf increased from 79% (2.37 of 3) to 96% (2.89 of 3) for legs with advanced peripheral vascular disease (PVD). Seventy-nine instead of 62 potential bypass recipients were identified (P = 0.002). Mean procedure time needed for selective catheterizations was 7 minutes. No adverse events were seen. CONCLUSIONS: Bilaterally antegrade selective DSA clearly is superior to aortic run-off DSA depicting tibial arteries. It requires comparatively small additional effort. Outflow vessel detection essentially is independent of advanced PVD. PMID- 8721958 TI - Computed tomography of experimental liver abscesses using a new liposomal contrast agent. AB - RATIONALE AND OBJECTIVES: Evaluation of computed tomographic enhancement characteristics of a new liposomal contrast agent (liposomal iodixanol [LI]) in a pyogenic liver abscess model in 17 rabbits. METHODS: Eight to 14 days after abscess induction (Escherichia coli), density-time curves were calculated for regions of interest in liver, abscess wall and center, spleen, portal vein, abdominal aorta, inferior vena cava, and kidney. Images were obtained every minute between 1 and 10 minutes, every 5 minutes between 15 and 60 minutes, and 75 minutes after 200 mg/kg LI application (group A: 7 rabbits) and after 600 mg/kg iopentol application (group B: 10 rabbits), and 90, 105, and 120 minutes after LI. RESULTS: The abscess wall-liver contrast after LI lasted from 10 to more than 120 minutes with a maximum of 30 delta Hounsfield Units (HU) at 45 minutes. For iopentol, the abscess wall-liver contrast lasted from 2 to 7 minutes with a maximum of 8 delta HU at 5 minutes. The abscess wall-center contrast after LI lasted from 1 to more than 120 minutes with a maximum of 112 delta HU at 40 minutes. For iopentol, the abscess wall-center contrast lasted from 1 to 75 minutes with a maximum of 79 delta HU at 1 minute. The liver-portal vein contrast after LI lasted from 1 to more than 120 minutes with a maximum of 100 delta HU at 20 minutes. For iopentol, the liver-portal vein contrast lasted from 1 to 8 minutes with a maximum of 38 delta HU at 2 minutes. An abscess wall was detected in a higher percentage of the LI images (86% LI, 56% iopentol), and images in the LI group correlated better with histopathology. CONCLUSIONS: The diagnostic value of LI exceeds that of iopentol in terms of overall abscess contrast and duration of the diagnostic interval. The higher hepatic vessel contrast allows better abscess localization. PMID- 8721959 TI - Reproducibility of human kidney perfusion and volume determinations with electron beam computed tomography. AB - RATIONALE AND OBJECTIVES: Alterations in whole kidney, cortical, and medullary perfusion and volume play a pivotal role in various physiologic and pathophysiologic processes. Electron-beam computed tomography (EBCT) provides accurate measurements of these traits in animals, but their reproducibility in humans has not been established. METHODS: Perfusion, volume, and flow measurements were obtained by EBCT in eight healthy human volunteers under controlled conditions on two consecutive days. RESULTS: Mean values for whole kidney, cortical, and medullary perfusion and volume obtained with EBCT were similar in scan 1 and scan 2 (P > 0.1), and correlated highly. Coefficients of variation for the repeated measurements usually were less than 10%. Values obtained for renal regional perfusion and volume agreed with previously reported values. CONCLUSIONS: Electron-beam computed tomography estimates of single whole kidney, cortical, and medullary perfusions and volumes are highly reproducible in normal humans, and may be useful to advance understanding of renal involvement in human disease. PMID- 8721960 TI - Hepatic kinetics and magnetic resonance imaging of gadolinium-EOB-DTPA in dogs. AB - RATIONALE AND OBJECTIVES: To measure the hepatic uptake and biliary elimination kinetics of gadolinium (Gd)-EOB-DTPA in dogs. METHOD: Two groups of four beagles each were anesthetized and given an intravenous bolus of 25 mumol/kg or 250 mumol/kg of Gd-EOB-DTPA. Blood, hepatic bile, and urine were collected over 140 minutes, and liver samples were obtained immediately after the dogs were killed. Conventional T1-weighted spin echo sequences of the liver were performed on a 1.5 Tesla (T) magnetic resonance imager during sampling. A ninth beagle received a bolus of 25 mumol/kg followed 140 minutes later with a bolus of 250 mumol/kg of Gd-EOB-DTPA. Wedge liver biopsies were obtained for Gd estimation at various times after dosing, and Gd concentration was measured by inductively coupled plasma atomic emission spectroscopy. RESULTS: The plasma concentration of Gd-EOB DTPA decreased in a biexponential manner with half-lives of approximately 4 minutes and 60 minutes for the distribution and elimination phase independent of the dose given. Gadolinium bile concentration reached peak values between 80 and 140 minutes: 6.3 +/- 1.6 mmol/L for the low dose (LD) and 11.6 +/- mmol/L for the high dose (HD). Bile Gd output was 62.0 +/- 8.8 (LD) and 78.3 +/- 30.2 (HD) nmol/minute-kg 50 to 80 minutes after injection. Gadolinium-EOB-DTPA was excreted by the biliary route to 24.8 +/- 2.6 (LD) and 3.6 +/- 1.2 (HD) percent of the dose within 140 minutes. Liver Gd concentration was 0.43 +/- 0.14 (LD) and 4.3 +/ 0.5 (HD) mmol/kg liver tissue at the conclusion of the studies. Calculated concentrations in the hepatocyte were 60 (LD) and 15 (HD) times higher than in plasma at 25 minutes after dosing. Whereas the low dose exhibited excellent contrast enhancement for the whole period, the high dose displayed a biphasic signal enhancement with a decreasing signal caused by the too-high hepatic gadolinium accumulation. CONCLUSIONS: Transport of the Gd-EOB-DTPA into the hepatocyte exceeded elimination from hepatocyte to bile. The high dose defined a biliary transport maximum for Gd-EOB-DTPA of 78.3 +/- 30.2 nmol/minute-kg. The liver accumulation results from fast transport into the hepatocyte and rate limited slower transport from hepatocyte to bile. The accumulation occurs against a strong concentration gradient, suggesting energy-dependent active transport into the hepatocyte. PMID- 8721961 TI - Behavior of bone mass measurements. Dual energy x-ray absorptiometry total body bone mineral content, ultrasound bone velocity, and computed metacarpal radiogrammetry, with age, gonadal status, and weight in healthy women. AB - RATIONALE AND OBJECTIVES: Bone mass as measured by dual energy x-ray absorptiometry varies with body weight. The authors studied the behavior of bone mass measurements made by ultrasound bone velocity and metacarpal radiogrammetry in relation to body weight. METHODS: Eighty healthy women were studied: 40 post menopausal and 40 premenopausal (mean age 60 +/- 6 and 38 +/- 8 years, respectively). The authors performed in every subject the following studies: a radiograph of the nondominant hand for metacarpal cortical thickness, a study of the 2nd to 5th proximal phalanges for ultrasound bone velocity, and total bone mineral content by dual energy x-ray absorptiometry. RESULTS: The measurements obtained with the three methods correlated significantly with each other (P < 0.0001). The only parameter related significantly with weight were total bone mineral content (P < 0.0001). In the premenopausal women, age did not correlate with any measurement. In the postmenopausal women, age correlated significantly with the three measured parameters (P < 0.0058 to P < 0.0001). CONCLUSIONS: Weight did not influence ultrasound bone velocity and metacarpal cortical thickness measurements. PMID- 8721962 TI - Covered stents for prevention of restenosis. Experimental and clinical results with different stent designs. AB - RATIONALE AND OBJECTIVES: Metallic stents in small vessels go along with a significant risk of restenosis and reocclusion. Different models of stents and covering materials have been purported to prevent intraluminal neointimal proliferation by cover-based closure of the spaces in the wire mesh. METHODS: Tantalum stents covered with polyethylacrylate/polymethylmethacrylate (PEM) were implanted in the infrarenal aorta of six New Zealand white rabbits by aortotomy and compared with eight rabbits treated with uncovered tantalum stents. For deployment, covered and uncovered stents necessitated a 7-French (F) and 5-F sheath, respectively. In addition, nine human patients with arteriosclerotic lesions of the superficial femoral arteries (stenosis > 5 cm or total occlusion) were treated percutaneously with a Dacron-covered nitinol vascular stent via a 9 F sheath. Patients were followed for a mean of 13.5 months, and control angiography was performed after 6 months. RESULTS: Experimental placement of the tantalum Wiktor stent was feasible technically in all cases. Five of six stents covered with PEM were occluded 3 days after placement despite the intravenous use of heparin and aspirin. In the group with uncovered stents, no area of stenosis greater than 10% was observed. There was a neointimal layer of 89 +/- 68 microns around the stent wires. Stent placement was successful in all patients. In four patients, a hyperergic reaction occurred, resulting in noninfectious periarteriitis. This complication was treated successfully with nonsteroidal antiinflammatory drugs. The primary patency was 50%, and the secondary patency (after application of a second covered stent in two patients) was 63%. CONCLUSIONS: The uncovered stent induces little neointimal proliferation around the stent wires. The insertion of stents covered with PEM into the rabbit aorta was accompanied by a strong thrombotic reaction, despite sufficient anticoagulation. Dacron-covered nitinol stents showed a surprisingly high restenosis rate after 9 months of follow-up. Further research concerning the in vivo properties of new covering materials is mandatory before routine vascular clinical application. PMID- 8721963 TI - Reliability of experimental models of iodinated contrast media-induced acute renal failure. From methodological considerations to pathophysiology. PMID- 8721964 TI - Accidental embryo irradiation during barium enema examinations. An estimation of absorbed dose. AB - RATIONALE AND OBJECTIVES: To investigate the possibility of an embryo to receive a dose of more than 10 cGy, the threshold of malformations induction in embryos reported by the International Commission on Radiological Protection, during barium enema examinations. METHODS: Thermoluminescent dosimeters were placed in a phantom to calculate the depth-to-skin conversion coefficient needed for dose estimation at the average embryo depth in patients. Barium enema examinations were performed in 20 women of childbearing age with diagnostic problems demanding longer fluoroscopy times. Doses at 6 cm, the average embryo depth, were determined by measurements at the patients' skin followed by dose calculation at the site of interest. RESULTS: The range of doses estimated at embryo depth for patients was 1.9 to 8.1 cGy. The dose always exceeded 5 cGy when fluoroscopy time was longer than 7 minutes. CONCLUSION: The dose at the embryo depth never exceeded 10 cGy. This study indicates that fluoroscopy time should not exceed 7 minutes in childbearing-age female patients undergoing barium enema examinations. PMID- 8721965 TI - Hemoglobin-nitric oxide interaction and its implications. PMID- 8721966 TI - Autologous fibrin glue: production and clinical use. PMID- 8721967 TI - von Willebrand disease: pathogenesis, classification, and management. PMID- 8721968 TI - The acquired-B phenomenon. PMID- 8721969 TI - Red cell compatibility testing: a perspective for the future. PMID- 8721970 TI - Blood proteins from transgenic animal bioreactors. PMID- 8721971 TI - The implementation and use of automated group and screen procedures in a hospital transfusion laboratory. PMID- 8721972 TI - The brain and the lymphatic system (I). PMID- 8721973 TI - The brain and the lymphatic system (II). PMID- 8721974 TI - Enzyme triple staining for differentiation of lymphatics from venous and arterial capillaries. AB - 5'-nucleotidase (5'-Nase)-dipeptidyl aminopeptidase IV (DAPase)-alkaline phosphatase (ALPase) triple staining was used to differentiate lymphatics from venous and arterial capillaries in a variety of mammalian tissue sections including human. This triple staining method facilitates specific identification under a light microscope of 5'-Nase activity in lymphatics, DAPase activity in venous capillaries and venules and ALPase activity in arterial capillaries and arterioles. This technique depicts initial lymphatics more clearly and extensively than other methods so far reported although some interspecies and tissue differences are obtained in each enzyme activity. PMID- 8721975 TI - Radical excision and delayed reconstruction of a lymphedematous leg with a 15 year follow-up. AB - An adult female patient with primary lymphedema of the lower extremity was treated with total excision of the subcutaneous tissues followed by delayed reconstruction with a giant full thickness skin graft taken from the excised surgical specimen. The leg has maintained excellent function and contour over the ensuing 15 years. A small area on the dorsum of the foot that initially was covered with a split thickness skin graft required subsequent regrafting using abdominal skin. This area developed verrucoid changes. PMID- 8721976 TI - P-selectin and von Willebrand factor in bovine mesenteric lymphatics: an immunofluorescent study. AB - P-selectin (PADGEM, GMP-140, CD62) is an integral membrane protein specific to alpha granules of platelets and Weibel-Palade bodies of blood vascular endothelial cells. The presence in lymphatic endothelial cells of numerous Weibel Palade bodies and their positivity to immunocytochemical reaction for von Willebrand factor have previously been characterized and described. Because von Willebrand factor and P-selectin codistribute in Weibel-Palade bodies of blood vascular endothelial cells we investigated the presence of both P-selectin and von Willebrand factor in lymphatic endothelium. Lymphatic vessels expressed positive reaction to immunocytochemical assay thereby demonstrating the presence of P-selectin in the endothelium. Distribution and intensity of the reaction were similar to those observed in bovine blood vascular endothelium. PMID- 8721977 TI - Treatment of secondary lymphedema of the upper limb with CYCLO 3 FORT. AB - Fifty seven patients with secondary lymphedema of the upper limb after previous treatment for breast cancer were treated for 3 months with an extract of Ruscus + Hesperidin Methyl Chalcone (CYCLO 3 FORT) or placebo according to a double-blind protocol in the context of a controlled clinical trial. All patients also underwent manual lymphatic drainage twice a week for at least one month. With CYCLO 3 FORT, the reduction in volume of arm edema, the main assessment criteria, was 12.9% after 3 months of treatment as compared with a placebo (p=0.009). Decreased edema tended to be more marked in the forearm compared with the upper arm where excess fat deposition seemed to dominate over excess fluid accumulation. CYCLO 3 FORT was well tolerated with minimal adverse reaction. PMID- 8721978 TI - Retroperitoneal laparoscopic lymph node dissection for staging non-seminomatous germ cell tumors before and after chemotherapy. AB - We describe our experience with laparoscopic retroperitoneal lymph node dissection in 19 patients with non-seminomatous germ cell tumors. Twelve patients had stage I disease with no clinical evidence (CT-scan, ultrasound, tumor markers) of metastases; 7 patients (stage IIb=2, stage IIc=5) had residual tumor after chemotherapy but with negative tumor markers. A laparoscopic dissection was used to asses more fully the pathologic status of the relevant retroperitoneal lymph nodes of both groups. The patient was positioned and trocars introduced at sites similar to that used for transperitoneal laparoscopic nephrectomy (flank position, five ports - 3 x 10 mm; 2 x 5 mm). After reflecting the colon anteromedially, the landmarks of the lymph node dissection were isolated-namely the ureter, aorta, inferior vena cava, and both renal veins. The lymph node dissection included the paracaval, interaorto-caval, upper preaortic, and right common iliac zonal nodes for right-sided tumors, and paraaortic, upper preaortic zones for left-sided tumors. Retrieval of the lymph nodal chains was accomplished using a small organ bag. The mean duration of the procedure was 298 (range 150 405) minutes. In only one patient was a lymph node positive for tumor (stage I). Otherwise nodes showed extensive necrosis (after chemotherapy). No intraoperative complications were encountered but three patients developed a delayed complication (ureteral stenosis, pulmonary embolism, and retrograde ejaculation, respectively). Whereas we completed the dissection in each patient with stage I tumors, the laparoscopic procedure was more difficult in patients with stage II tumors after chemotherapy. In two patients with stage IIb disease laparoscopic lymphadenectomy was successful. In four other patients parts of the dissection had to be done after conversion to an open (conventional) operation using a small incision (suprainguinal or pararectal); in one patient the laparoscopic approach was abandoned and converted to an open operation. In the post-chemotherapy group the outcome depended primarily on the tumor bulk prior to drug treatment. In two patients in whom all residual necrotic tissue was removed laparoscopically they had "minor" disease (stage IIb); the others had stage IIc tumors. Our preliminary experience suggests that a modified laparoscopic lymph node dissection is feasible for stage I tumors and in selected patients with marker negative residual tumor after chemotherapy (stage IIb). PMID- 8721979 TI - Evidence for redundancy in propeptide/prohormone convertase activities in processing proglucagon: an antisense study. AB - To further examine the physiological roles of the neuroendocrine prohormone convertases (PCs) in proglucagon processing, alpha TC1-6 cells were transiently transfected with PC1/3 and PC2 expression vectors containing either antisense or sense encoding cDNAs. PC1/3- and PC2-directed RIAs were used to determine that the PC1/3 antisense transfections lowered endogenous levels of PC1/3 by 40 +/- 7.9% but did not alter the levels of PC2. The PC2 antisense transfections decreased the endogenous levels of PC2 by 91 +/- 11.7% without affecting the levels of PC1/3. To quantitate the levels of proglucagon and proglucagon-derived products, transfected cells were metabolically labeled with [3H]tryptophan, and extracts were chromatographed by reversed-phase HPLC. Recovered peptides were then subjected to peptide mapping analyses, allowing precise quantification of 3H radioactivity incorporated into proglucagon and its cleavage products. Product precursor ratios were determined, and percent change in the proportion of products generated in antisense-transfected vs. sense-transfected cells was calculated. The decrease in PC1/3 after antisense treatment significantly reduced the amounts of glicentin produced and partially reduced the levels of all other proglucagon cleavage products. PC2 antisense treatment significantly reduced the levels of glicentin and 9K glucagon generated but had no significant effect on the remainder of the proglucagon-derived peptides. These results suggest the existence of redundant mechanisms that ensure the production of each of the intermediate and product peptides derived from proglucagon. PC1/3 is potentially an important enzyme in the processing of most proglucagon-derived peptides, whereas PC2-processing activity appears to predominate at only two of the four potential cleavage sites. PMID- 8721980 TI - Role of prohormone convertases in the tissue-specific processing of proglucagon. AB - Proglucagon (proG) is processed in a tissue-specific manner to glucagon in the pancreas and to gilcentin, oxyntomodulin, glucagon-like peptide (GLP)-1, and GLP 2 in the intestine. Recombinant vaccinia virus (vv) vectors were used to infect prohormone convertase 1 (PC1) or PC2 into nonendocrine (BHK-proG) cells, which stably express proG. Similarly, endocrine (GH3, AtT-20) cells were coinfected with proG along with PC1 or PC2 alone, or in combination with furin, PACE4, PC5a, or PC5b. Cell extracts were analyzed for various proG-derived peptides by RIA of fractions obtained from HPLC. Upon infection of BHK-proG cells with either vv: furin or vv:PC1, glicentin was produced, while vv: PC2 did not process proG. In GH3 and AtT-20 cells, vv:PC1 produced glicentin, oxyntomodulin, GLP-1(1-37), GLP 1(7-37), and GLP-2. All other enzymes tested produced only glicentin. Interestingly, no enzyme or combination produced glucagon. Coinfection of GH3 cells with vv:PC2 and members of the chromogranin family of peptides, including chromogranin A and B and secretogranin II, as well as the PC2-binding protein 7B2, did not result in processing to glucagon. It is concluded that: 1) PC1 is responsible for the processing of proG to produce the intestinal peptides glicentin, oxyntomodulin, GLP-1(1-37), GLP-1(7-37), and GLP-2, and 2) PC2 processes proG to glicentin but does not produce glucagon, alone or in combination with other enzymes or with known molecular chaperones. PMID- 8721981 TI - Transcriptional activation of the gonadotropin-releasing hormone receptor gene by activin A. AB - It has been reported that activin A stimulates the synthesis of the GnRH receptors (GnRHR) in rat pituitary cultures. However, the role of activin A in the regulation of the GnRHR gene at the molecular level is not known. In the present work, we have studied the regulation of the GnRHR gene by activin A in the gonadotrope cell line, alpha T3-1, where the GnRHR gene is highly expressed. First, we demonstrate that these cells express the mRNAs of three types of activin receptors: I, II, and IIB. Activin A increases GnRHR mRNA levels in a dose-and time-dependent manner, with maximal stimulation (2.5 +/- 0.5-fold) occurring with a dose of 20 ng/ml after 36 h of incubation. To ascertain whether this effect occurs at the transcriptional level, we performed nuclear run-off experiments in alpha T3-1 cells, which demonstrate a 1.6-fold increase in the levels of newly synthesized GnRHR mRNA in response to activin A. To investigate further the effect of activin A on the transcription of the GnRHR gene, alpha T3 1 cells were transiently transfected with a mouse GnRHR promoter/luciferase reporter gene (GnRHR-Luc) and challenged with activin A. Luciferase activity increases in response to activin A to the same extent (2.4 +/- 0.4-fold) and with similar dose-response and time-course profiles as the mRNA levels. Follistatin (100 ng/ml), a well known activin antagonist, completely abolishes the activin A effect on both mRNA levels and GnRHR-Luc activity. Follistatin also decreases the basal expression of the GnRHR gene by 33% as determined by GnRHR-Luc activity. This, together with our demonstration of the presence of the inhibin beta B subunit mRNA in alpha T3-1 cells, suggests a potential paracrine/autocrine role of endogenous activin B on the regulation of the GnRHR gene in these cells. To provide evidence for biological significance of activin A stimulation of GnRHR gene expression, the response of a human gonadotropin alpha-subunit promoter/luciferase reporter gene (alpha Gon-Luc) to GnRH was assessed in alpha T3-1 cells pretreated with activin A. Activin enhances the stimulation of alpha Gon-Luc activity by GnRH by 1.6 +/- 0.4-fold. These data demonstrate that activin A can stimulate the expression of the GnRHR gene at the transcriptional level. Furthermore, transfection studies localize the activin responsive element to 1.2 kb of the 5'-flanking region of the GnRHR gene. Transcriptional activation of the GnRHR gene by activin A may serve as a mechanism for the modulation of gonadotrope responsiveness to GnRH. PMID- 8721982 TI - Formation and activation by phosphorylation of activin receptor complexes. AB - Activin is a protein growth and differentiation factor that initiates intracellular events through the activation of a complex of transmembrane protein serine kinases. Two subfamilies of receptor serine kinases, type I and type II, have been identified, and both receptor types may be required to generate a transmembrane signal. Investigation of the interaction between various activin receptors (ActRs) revealed that ActRs I and II could exist in a stable complex and that formation of that complex between transiently overexpressed molecules was not regulated by ligand. Analysis of phosphorylation suggested that activin induced phosphorylation of receptor I, probably at residues within a conserved glycine and serine-rich sequence in the juxtamembrane region referred to as the GS domain. Phosphorylation of the GS domain was dependent upon a functional ActRII. Introduction of an activin type I receptor, ALK4, into the mink lung epithelial cell line, L17, conferred activin responsiveness on those cells. Mutation of specific combinations of serines and threonines in the core sequence of the ALK4 GS domain to alanine rendered that receptor incompetent for signaling. Mutation of the same sets of residues to glutamic acid produced molecules that supported activin signaling but that did not display elevated basal signaling anticipated for a constitutively active receptor. However, mutation of a threonine residue in the carboxy-terminal half of the GS domain, T206, to glutamic acid yielded receptors with constitutive activity. Taken together, these results support a role for phosphorylation of type I ActRs in the generation of a biological signal. PMID- 8721983 TI - 5 alpha-reduced androgens play a key role in murine parturition. AB - Two steroid 5 alpha-reductase isozymes designated type 1 and 2 synthesize 5 alpha reduced androgens and other 5 alpha-reduced steroid hormones. Naturally occurring mutations in the gene encoding 5 alpha-reductase type 2 cause male pseudohermaphroditism, indicating that this isozyme is responsible for the synthesis of dihydrotestosterone required for virilization of the embryonic male urogenital tract. To determine the physiological role of 5 alpha-reductase type 1, homologous recombination in mouse embryonic stem cells was used to produce male and female mice with a disruption (null allele) in the type 1 gene (Srd5a1). Male mice lacking 5 alpha-reductase type 1 appear normal. Females exhibit a parturition defect that is maternal in origin. The parturition defect is reversed by administration of 5 alpha-androstan-3 alpha, 17 beta-diol (3 alpha-Adiol), a 5 alpha-reduced androgen previously thought to be a breakdown product. Enzymes that synthesize 3 alpha-Adiol, including 5 alpha-reductase type 1 and 3 alpha hydroxysteroid dehydrogenase, are induced in wild type uterus during late gestation. Induction leads to peak circulating levels of 3 alpha-Adiol on days 17/18 of gestation in wild type but not mutant mice. The results document a role for 5 alpha-reduced androgens synthesized by the type 1 isozyme in normal female physiology, and they suggest that 3 alpha-Adiol is a new hormone required for parturition in mice. PMID- 8721984 TI - Progesterone receptor-induced bending of its target DNA: distinct effects of the A and B receptor forms. AB - We have used circular permutation and phasing electrophoretic mobility shift assays to determine the ability of the A and B forms of human progesterone receptor (PR) to bend target DNA. Studies were done with baculovirus-expressed full-length receptors purified to apparent homogeneity. By circular permutation analysis, both forms of PR induced substantial distortions in the structure of target DNA with calculated distortion angles (alpha D) of 57 degrees for PR-A and 84 degrees for PR-B. The apparent bend centers for both forms of PR were similarly located a few base pairs (-4 to -2 bp) from the middle of the progesterone response element. No differences were detected in the magnitude of distortion or apparent bend centers when PR was bound to hormone agonist (R5020) or the antagonist RU486. Phasing analysis, which can determine the orientation of a DNA bend, revealed that both forms of PR mediated directional bends toward the major groove of the DNA helix. Calculated directed bend angles (alpha B) were 40 degrees for PR-B and 31 degrees for PR-A. The chromatin high mobility group protein HMG-1, which acts as an accessory factor to enhance the binding affinity of purified PR for progesterone response elements, had minimal influence on PR mediated DNA bending. This result, taken together with the fact that HMG-1 can form a ternary complex with PR and DNA, is consistent with the conclusion that HMG-1 facilitates PR binding by stabilizing a receptor-induced DNA conformation that is required for assembly of a high affinity PR-DNA complex. The results of this study also suggest that DNA bending may be coupled to transcriptional regulation since PR-B is generally a stronger transcriptional activator than PR-A and also mediates a larger bend in target DNA than PR-A. PMID- 8721985 TI - Retinoid X receptor isotype identity directs human vitamin D receptor heterodimer transactivation from the 24-hydroxylase vitamin D response elements in yeast. AB - Unlike estrogen and progesterone receptors that operate as homodimers on response elements, retinoid X receptors (RXRs) and vitamin D receptors (VDRs) can function as heterodimers. Studies concerning the significance of heterodimeric partnerships are usually performed utilizing mammalian or insect cells. These cells express endogenous nuclear receptors, making it impossible to assign a role for one receptor subtype over another while studying the function of transfected receptor(s). Yeast lacks endogenous VDRs and RXRs and their ligands and provides a unique cellular context to study nuclear receptor function. We examined the interaction between human VDR and human RXR alpha, mouse RXR beta 2, and mouse RXR gamma to identify physiologically important receptor interactions. DNA binding studies on consensus, osteocalcin, or the rat 24-hydroxylase vitamin D response elements (VDREs) indicated that although RXR complexes can form on the consensus DNA elements, RXR:VDR heterodimers preferentially interact with the natural VDREs. The interaction is RXR isotype-specific and affected by ligands. Transactivation studies using the rat 24-hydroxylase VDREs indicated that VDR preferentially associated with RXR alpha or RXR gamma to stimulate transcription, and the activity was potentiated by ligand. Although RXR beta 2:VDR bound tightly to DNA, the resulting heterodimer transactivated poorly. The regulation of the 24 hydroxylase promoter observed in yeast is similar with respect to transactivation potential of specific VDRE and fold activation observed in osteosarcoma cells. Ligand binding to both receptors in a RXR:VDR complex is required for maximal transcriptional activity, indicating that the isotype-specific RXR partner significantly contributes to the ability of RXR:VDR heterodimers to transactivate from target response elements in yeast. PMID- 8721986 TI - Molecular cloning of human p48, a transient component of progesterone receptor complexes and an Hsp70-binding protein. AB - A 48-kDa protein (p48) that transiently associates with progesterone receptor during cell-free assembly in rabbit reticulocyte lysate was isolated by two dimensional gel separation. Tryptic peptide sequences were generated and used to develop an antipeptide antiserum recognizing p48 by Western immunostaining, and this antiserum was used to monitor purification of native p48 from reticulocyte lysate. Eight mouse monoclonal antibodies capable of immunoprecipitating vertebrate p48 were generated. These monoclonal antibodies served as probes to clone ten p48 cDNAs from a HeLa cDNA expression library. One of the cloned cDNAs was sequenced in its entirety and codes for a 369-amino acid protein (calculated Mr = 41,324). Northern blot analysis of RNA from multiple human tissues suggest that p48 may be ubiquitously expressed. Expression of human p48 cDNA in vitro yielded a product that comigrated with rabbit p48 by SDS-PAGE and associated with progesterone receptor in a similar manner. Immunoprecipitation of p48 complexes revealed a common association of p48 with hsp70 and, to a lesser extent, with hsp90 and p60. Thus, it appears that p48 is a novel component of the cytoplasmic molecular chaperone machinery. PMID- 8721987 TI - Protein kinase C-dependent growth hormone releasing peptides stimulate cyclic adenosine 3',5'-monophosphate production by human pituitary somatotropinomas expressing gsp oncogenes: evidence for crosstalk between transduction pathways. AB - The effects of the synthetic GH-releasing peptides, GHRP-2 and GHRP-6, on phosphatidylinositol (PI) hydrolysis and cAMP production have been examined in human pituitary somatotropinomas with and without adenylyl cyclase-activating gsp oncogenes. Both peptides dose-dependently stimulated the rate of PI hydrolysis and GH secretion by cell cultures of both types of somatotropinoma. GHRP-2 was considerably more potent than GHRP-6. The effects on GH secretion were reduced or abolished by phloretin, an inhibitor of protein kinase C, and W7, an inhibitor of calmodulin. However, antagonism of the GHRH-receptor and of protein kinase A with (N-Ac-Tyr1,D-Arg2)GRF-(1-29)-NH2 and Rp-adenosine-3',5'-cyclic monophosphothioate, respectively, did not alter the stimulatory effects of GHRP-2 and GHRP-6 on GH secretion. The effect of GHRP-2 and/or GHRP-6 on cAMP production was studied in 15 tumors, seven of which possessed constitutive adenylyl cyclase activity as evidenced by presence of gsp oncogenes. Both peptides stimulated cAMP production in the latter but not former types of tumor. Moreover, GHRP-2 and GHRP 6 potentiated the stimulation of cAMP production induced by GHRH and pituitary adenylate cyclase-activating polypeptide in tumors without gsp oncogenes. These results demonstrate that GHRP-2 and GHRP-6 exert identical effects on human pituitary somatotropinomas, except for differences in potency. Additionally, under conditions of adenylyl cyclase activity above basal levels (i.e. through stimulation of G2-protein coupled receptors or because of gsp oncogene expression), cAMP production can be increased even further by GHRP, providing evidence for cross-talk between the PI and adenylyl cyclase transduction systems in pituitary cells. PMID- 8721988 TI - Steroid and pulsatile gonadotropin-releasing hormone (GnRH) regulation of luteinizing hormone and GnRH receptor in a novel gonadotrope cell line. AB - Properties of a pituitary gonadotrope include the capacity to regulate gonadotropin synthesis and secretion in response to a GnRH signal. Progress in identifying the steps involved in these processes has been impeded by the lack of a homogeneous in vitro model of gonadotropes. This paper presents functional characterization of a L beta T2 gonadotrope cell line generated by tumorigenesis in transgenic mice carrying the rat LH beta-subunit regulatory region linked to the SV40 T-antigen oncogene. This cell line expresses LH beta, alpha-subunit, and GnRH-receptor (GnRH-R) mRNAs (though not FSH beta), responds to glucocorticoid treatment with a reversible dampening of proliferation, and responds to pulsatile, concentration-dependent GnRH administration with LH secretion. L beta T2 cells presented with four GnRH pulses (10 nM, 90-min interpulse interval) on each of 4 days respond with incremental increases in LH secretion on successive days. This increase was greatest (15-fold) in the presence of estradiol and dexamethasone. Part of the enhanced responsiveness is apparently due to an increase in GnRH-R; pulsatile GnRH treatment alone as well as steroid treatment alone led to an increase in GnRH-R mRNA levels. When secretion was stimulated on day 4 with 54 mM [K+] pulses, bypassing the GnRH-R, the LH-secretory response indicated that the GnRH pulse history as well as estradiol and dexamethasone have actions on L beta T2-secretory capacity distinct from changes in the GnRH-R. This increase can be explained in part by the marked up-regulation of LH beta, but not alpha-subunit, mRNA observed in GnRH-pulsed cells. In summary, L beta T2 clonal gonadotropes exhibit functional characteristics consistent with those of normal pituitary gonadotropes such as LH secretion via a regulated pathway and changes in GnRH-R and LH beta gene expression in response to signaling by GnRH and steroid hormones and therefore should be a useful tool for dissecting the cellular and molecular events involved in these fundamental gonadotrope properties. PMID- 8721989 TI - Convergence of signaling transduced by prolactin (PRL)/cytokine chimeric receptors on PRL-responsive gene transcription. AB - Ligand binding to cytokine receptors rapidly triggers tyrosine phosphorylation of Janus family tyrosine kinases (Jaks) and signal transducers and activators of transcription (Stats). Jak2 activation is mediated by PRL receptor homodimers as well as by receptors for the interleukin (IL)-3, IL-5, and granulocyte macrophage colony stimulating factor, which share the common beta c-subunit. Otherwise, Jak1 and Jak3 are involved in IL-2 signaling through heterodimerization of the IL-2 receptor-beta (IL-2R beta) and gamma c-chains. Stat5, a member of the Stat family, confers the PRL response on milk protein genes. Here we show that chimeric PRL receptors that contain the transmembrane and cytoplasmic domains of the IL-2R beta or beta c-chains transduce in response to PRL tyrosine phosphorylation and activation of Jak1 and Jak2, respectively. Tyrosine phosphorylation of Stat5, activation of its DNA-binding activity assessed in bandshift experiments using a lactogenic hormone responsive region (LHRR) probe, and transcriptional induction of a beta-casein promoter luciferase construct in stably transfected CHO cells are observed with both chimeras upon PRL stimulation. Our results demonstrate that distinct cytoplasmic domains of these cytokine receptors elicit convergent signaling pathways and provide evidence that beta c and IL-2R beta function as a complete signal transducer. Our data strengthen previous observations that Stat5 activation is not dependent on the activation of a specific Jak kinase and also suggest that neither Jak3 nor gamma c have a specific role in this process. PMID- 8721990 TI - A milestone in ribosomal crystallography: the construction of preliminary electron density maps at intermediate resolution. AB - Preliminary electron density maps of the large and the small ribosomal particles from halophilic and thermophilic sources, phased by the isomorphous replacement method, have been constructed at intermediate resolution. These maps contain features comparable in size with what is expected for the corresponding particles, and their packing arrangements are in accord with the schemes obtained by ab-initio procedures as well as with the motifs observed in thin sections of the crystals by electron microscopy. To phase higher resolution data, procedures are being developed for derivatization by specific labeling of the ribosomal particles at selected locations with rather small and dense clusters. Potential binding sites are being inserted either by site directed mutagenesis or by chemical modifications to facilitate cluster binding on the surface of the halophilic large and the thermophilic small ribosomal particles, which yield the crystals diffracting to highest resolution (2.9 and 7.3 A (1 A = 0.1 nm), respectively). For this purpose, the surface of these ribosomal particles is being characterized and procedures are being developed for quantitative detachment of selected ribosomal proteins and for their incorporation into core particles. The genes of these proteins are being cloned, sequenced, mutated to introduce reactive side groups, mainly cysteines, and overexpressed. In parallel, two in situ small and stable complexes were isolated from the halophilic ribosome. Procedures for their crystal production in large quantities are currently being developed. Models, reconstructed at low resolution from crystalline arrays of ribosomes and their large subunits, are being used for initial low-resolution phasing of the X-ray amplitudes. The interpretation of these models stimulated the design and the crystallization of complexes mimicking defined functional states of a higher quality than those obtained for isolated ribosomes. These models also inspired modelling experiments according to results of functional studies, performed elsewhere, focusing on the progression of nascent proteins. PMID- 8721991 TI - Modeling the structure of the ribosome. AB - Considering the size and complexity of the ribosome and the growing body of data from a wide range of experiments on ribosomal structure, it is becoming increasingly important to develop tools that facilitate the development of reliable models for the ribosome. We use a combination of manual and computer based approaches for building and refining models of the ribosome and other RNA protein complexes. Our methods are aimed at determining the range of models compatible with the data, making quantitative statements about the positional uncertainties (resolution) of different regions, identifying conflicts in the data, establishing which regions of the ribosome need further experimental exploration, and, where possible, predicting the outcome of future experiments. Our previous low-resolution model for the small subunit of the Escherichia coli ribosome is briefly reviewed, along with progress on atomic resolution modeling of the mRNA-tRNA complex and its interaction with the decoding site of the 16S RNA. PMID- 8721992 TI - A model of the translational apparatus based on a three-dimensional reconstruction of the Escherichia coli ribosome. AB - The morphology of the Escherichia coli ribosome, i.e., its shape at moderate to low (20-40 A (1 A = 0.1 nm)) resolution, provides important constraints in modeling both the folding of ribosomal RNA and the translational process. A new reconstruction, obtained by low-dose cryoelectron microscopy and image processing of single ribosomes, contains clues to the way in which the ribosome interacts with the key functional ligands: the mRNA and the A- and P-site tRNAs. It also suggests possible pathways of the nascent polypeptide chain. From an interpretation of these clues in the light of existing knowledge, a plausible model for the locations and interactions of key components of protein synthesis is suggested. PMID- 8721993 TI - Getting closer to an understanding of the three-dimensional structure of ribosomal RNA. AB - Two experimentally unrelated approaches are converging to give a first low resolution solution to the question of the three-dimensional organization of the ribosomal RNA from Escherichia coli. The first of these is the continued use of biochemical techniques, such as cross-linking, that provide information on the relative locations of different regions of the RNA. In particular, recent data identifying RNA regions that are juxtaposed to functional ligands such as mRNA or tRNA have been used to construct improved topographical models for the 16S and 23S RNA. The second approach is the application of high-resolution reconstruction techniques from electron micrographs of ribosomes in vitreous ice. These methods have reached a level of resolution at which individual helical elements of the ribosomal RNA begin to be discernible. The electron microscopic data are currently being used in our laboratory to refine the biochemically derived topographical RNA models. PMID- 8721994 TI - Genomic evolution drives the evolution of the translation system. AB - Our thesis is that the characteristics of the translational machinery and its organization are selected in part by evolutionary pressure on genomic traits have nothing to do with translation per se. These genomic traits include size, composition, and architecture. To illustrate this point, we draw parallels between the structure of different genomes that have adapted to intracellular niches independently of each other. Our starting point is the general observation that the evolutionary history of organellar and parasitic bacteria have favored bantam genomes. Furthermore, we suggest that the constraints of the reductive mode of genomic evolution account for the divergence of the genetic code in mitochondria and the genetic organization of the translational system observed in parasitic bacteria. In particular, we associate codon reassignments in animal mitochondria with greatly simplified tRNA populations. Likewise, we relate the organization of translational genes in the obligate intracellular parasite Rickettsia prowazekii to the processes supporting the reductive mode of genomic evolution. Such findings provide strong support for the hypothesis that genomes of organelles and of parasitic bacteria have arisen from the much larger genomes of ancestral bacteria that have been reduced by intrachromosomal recombination and deletion events. A consequence of the reductive mode of genomic evolution is that the resulting translation systems may deviate markedly from conventional systems. PMID- 8721995 TI - Processing of eukaryotic pre-rRNA: the role of the transcribed spacers. AB - The 17-18S, 5.8S, and 25-28S rRNA species of eukaryotic cells are produced by a series of nucleolytic reactions that liberate the mature rRNAs from the large primary precursor transcript synthesized by RNA polymerase 1. Whereas the order of the cleavage reactions has long been established, until recently little information was available on their molecular details, such as the nature of the proteins, including the nucleolytic enzymes, involved and the signals directing the processing machinery to the correct sites. This situation is now rapidly changing, in particular where yeast is concerned. The use of recently developed systems for in vivo mutational analysis of yeast rDNA has considerably enhanced our knowledge of cis-acting structural features within the pre-rRNA, in particular the transcribed spacer sequences, that are critical for correct and efficient removal of these spacers. The same systems also allow a link to be forged between trans-acting processing factors and these cis-acting elements. In this paper, we will focus predominantly on the nature and role of the cis-acting processing elements as identified in the transcribed spacer regions of Saccharomyces cerevisiae pre-rRNA. PMID- 8721996 TI - Trans-acting factors in yeast pre-rRNA and pre-snoRNA processing. AB - The major intermediates in the pathway of pre-rRNA processing in yeast and other eukaryotes were originally identified by biochemical analyses. However, as a result of the analysis of the effects of mutations in trans-acting factors, the yeast pre-rRNA processing pathway is now characterized in far more detail than that of other eukaryotes. These analyses have led to the identification of processing sites and intermediates that were either too close in size or too short lived to detected by biochemical analyses alone. In addition, it was generally unclear whether pre-rRNA processing steps were endonucleolytic or exonucleolytic; analyses of trans-acting factors is now revealing a complex mixture of endonucleolytic and exonucleolytic processing steps. Many of the small nucleolar RNAs (snoRNAs) are excised from larger precursors. Analyses of trans acting factors are also revealing details of pre-snoRNA processing in yeast. Interestingly, factors involved in pre-snoRNA processing turn out to be components that also function in pre-rRNA processing, suggesting a potential mechanism for the coregulation of rRNA and snoRNA synthesis. In general, very little is known about the regulation of pre-rRNA processing steps. The best candidate for a system regulating specific pre-rRNA processing reactions has recently been revealed by the analysis of a yeast pre-RNA methylase. Here we will review recent data on the trans-acting factors involved in yeast ribosome synthesis and discuss how these analyses have contributed to our current view of this complex process. PMID- 8721997 TI - Preribosomal RNA processing in archaea: characterization of the RNP endonuclease mediated processing of precursor 16S rRNA in the thermoacidophile Sulfolobus acidocaldarius. AB - The hyperthermophilic archaeon Sulfolobus acidocaldarius uses a novel RNA containing endonuclease to excise and mature 16S rRNA from the precursor (pre) rRNA transcript. A cell-free processing system has been developed using an in vitro transcribed RNA substrate containing the entire 144 nucleotide 5' external transcribed spacer (5'ETS) and the first 72 nucleotides of 16S rRNA. The cell free extract cleaves in the 5'ETS at positions -99, -31, and +1 (i.e., the 5'ETS 16S junction). These positions are at or near the positions cleaved in vivo during processing of the pre rRNA transcript. The processing activity has been purified between 100 and 200-fold and appears to contain five or six polypeptide components and perhaps as many as 10 different small RNA components. Using combined reverse transcription-PCR amplification, full or partial cDNA copies of two of the RNA components have been obtained. One of the RNAs exhibits sequence and structural similarities to eukaryotic U3 snoRNA. The processing activity has been shown to be inactivated by micrococcal nuclease. It can be reactivated by reconstituting using bulk RNA from S.acidocaldarius but not bulk RNA from distantly related organisms. The activity is also abolished by RNase H digestion in the presence of oligonucleotides complementary to the U3-like RNA. These results demonstrate that the U3-like RNA is an essential component of the pre rRNA processing RNP endonuclease. Furthermore, this RNP endonuclease is not a derived eukaryotic feature, instead its existence predates the divergence of archaea and eukaryotes. PMID- 8721998 TI - Global regulators of ribosome biosynthesis in yeast. AB - Three abundant ubiquitous DNA-binding protein factors appear to play a major role in the control of ribosome biosynthesis in yeast. Two of these factors mediate the regulation of transcription of ribosomal protein genes (rp-genes) in yeasts. Most yeast rp-genes are under transcriptional control of Rap1p (repressor activator protein), while a small subset of rp-genes is activated through Abf1p (ARS binding factor). The third protein, designated Reb1p (rRNA enhancer binding protein), which binds strongly to two sites located upstream of the enhancer and the promoter of the rRNA operon, respectively, appears to play a crucial role in the efficient transcription of the chromosomal rDNA. All three proteins, however, have many target sites on the yeast genome, in particular, in the upstream regions of several Pol II transcribed genes, suggesting that they play a much more general role than solely in the regulation of ribosome biosynthesis. Furthermore, some evidence has been obtained suggesting that these factors influence the chromatin structure and creat a nucleosome-free region surrounding their binding sites. Recent studies indicate that the proteins can functionally replace each other in various cases and that they act synergistically with adjacent additional DNA sequences. These data suggest that Abf1p, Rap1p, and Reb1p are primary DNA-binding proteins that serve to render adjacent cis-acting elements accessible to specific trans-acting factors. PMID- 8721999 TI - Novel intron-encoded small nucleolar RNAs with long sequence complementarities to mature rRNAs involved in ribosome biogenesis. AB - Recently, several new snoRNAs encoded in introns of genes coding for ribosomal, ribosome-associated, or nucleolar proteins have been discovered. We are presently studying four of these intronic snoRNAs. Three of them, U20, U21, and U24, are closely related to each other on a structural basis. They are included in genes encoding nucleolin and ribosomal proteins L5 and L7a, respectively, in warm blooded vertebrates. These three metabolically stable snoRNAs interact with nucleolar protein fibrillarin. In addition, they display common features that make them strikingly related to snoRNA U14. U14 contains two tracts of complementarity to 18S rRNA, which are required for the production of 18S rRNA. U20 displays a 21 nucleotide (nt) long complementarity to 18S rRNA. U21 contains a 13 nt complementarity to an invariant sequence in eukaryotic 28S rRNA. U24 has two separate 12 nt long complementarities to a highly conserved tract of 28S rRNA. Phylogenetic evidences support the fundamental importance of the pairings of these three snoRNAs to pre-rRNA, which could be involved in a control of pre rRNA folding during preribosome assembly. By transfection of mouse cells, we have also analyzed the processing of U20 and found that the -cis acting signals for its processing from intronic RNA are restricted to the mature snoRNA sequence. Finally, we have documented changes of host genes for these three intronic snoRNAs during the evolution of eukaryotes. PMID- 8722000 TI - Small nucleolar RNA. AB - A growing list of small nucleolar RNAs (snoRNAs) has been characterized in eukaryotes. They are transcribed by RNA polymerase II or III; some snoRNAs are encoded in the introns of other genes. The nonintronic polymerase II transcribed snoRNAs receive a trimethylguanosine cap, probably in the nucleus, and move to the nucleolus. snoRNAs are complexed with proteins, sometimes including fibrillarin. Localization and maintenance in the nucleolus of some snoRNAs requires the presence of initial precursor rRNA (pre-rRNA). Many snoRNAs have conserved sequence boxes C and D and a 3' terminal stem; the role of these features are discussed. Functional assays done for a few snoRNAs indicate their roles in rRNA processing for cleavage of the external and internal transcribed spacers (ETS and ITS). U3 is the most abundant snoRNA and is needed for cleavage of ETS1 and ITS1; experimental results on U3 binding sites in pre-rRNA are reviewed. 18S rRNA production also needs U14, U22, and snR30 snoRNAs, whereas U8 snoRNA is needed for 5.8S and 28S rRNA production. Other snoRNAs that are complementary to 18S or 28S rRNA might act as chaperones to mediate RNA folding. Whether snoRNAs join together in a large rRNA processing complex (the "processome") is not yet clear. It has been hypothesized that such complexes could anchor the ends of loops in pre-rRNA containing 18S or 28S rRNA, thereby replacing base-paired stems found in pre-rRNA of prokaryotes. PMID- 8722002 TI - Structure and function of 5S rRNA in the ribosome. AB - 5S rRNA is a small RNA molecule that is a component of a ribosome from almost all living organisms. In this review, we discuss the biogenesis of 5S rRNA and its properties as an independent structural domain of a ribosome as well as the current concepts concerning the higher order structure of 5S rRNA in free state and in its complexes with ribosomal proteins and its folding in the ribosome. Special attention is paid to recent experimental approaches that have been useful in 5S rRNA studies. Our own data on topography of 5S rRNA in the ribosomes are discussed in detail. The hypothesis describing the possible functional role of 5S rRNA for ribosome functioning is discussed. PMID- 8722003 TI - Antibiotic inhibition of the movement of tRNA substrates through a peptidyl transferase cavity. AB - The present review attempts to deal with movement of tRNA substrates through the peptidyl transferase centre on the large ribosomal subunit and to explain how this movement is interrupted by antibiotics. It builds on the concept of hybrid tRNA states forming on ribosomes and on the observed movement of the 5' end of P site-bound tRNA relative to the ribosome that occurs on peptide bond formation. The 3' ends of the tRNAs enter, and move through, a catalytic cavity where antibiotics are considered to act by at least three primary mechanisms: (i) they interfere with the entry of the aminoacyl moiety into the catalytic cavity before peptide bond formation; (ii) they inhibit movement of the nascent peptide along the peptide channel, a process that may generally involve destabilization of the peptidyl tRNA, and (iii) they prevent movement of the newly deacylated tRNA between the P/P and hybrid P/E sites on peptide bond formation. PMID- 8722001 TI - Genetic probes of ribosomal RNA function. AB - We have used a genetic approach to uncover the functional roles of rRNA in protein synthesis. Mutations were constructed in a cloned rrn operon by site directed mutagenesis or isolated by genetic selections following random mutagenesis. We have identified mutations that affect each step in the process of translation. The data are consistent with the results of biochemical and phylogenetic analyses but, in addition, have provided novel information on regions of rRNA not previously investigated. PMID- 8722004 TI - A proposal for the conformation of loop E in Escherichia coli 5S rRNA. AB - A proposal is advanced for the conformation of the loop E region of prokaryotic 5S rRNAs based on spectroscopic data obtained from pAD3 RNA, a construct that includes helix IV, helix V, and loops D and E from Escherichia coli 5S rRNA. Even though loop E juxtaposes bases that cannot form Watson-Crick base pairs, it resembles an A-form double helix; its nucleotides relate to each other spectroscopically in a helix-like way and are in the anti conformation. The ends of loop E, which is palindromic, have the same conformation. Working in from either end towards the center of the loop, a closing GC is followed by a side-by side GA and then by a reversed Hoogsteen AU, a pattern resembling that found at one end of eukaryotic loop E. The center of the loop consists of three nucleotide pairs, which appear to be an asymmetric GG pair, a Watson-Crick-like AG, and a GU stabilized by a single hydrogen bond. PMID- 8722005 TI - An oligonucleotide analog approach to the decoding region of 16S rRNA. AB - Despite the passage of about 30 years since the discovery of the translational activities of ribosomes and the outlining of the roles of the large and small subunits, the actual molecular basis for the mRNA decoding activities of the small subunit has remained essentially obscure. In this paper, we describe a new approach using oligonucleotide analogs of 16S ribosomal RNA, in which the small ribosomal subunit is effectively deconstructed into a smaller more experimentally tractable form. Specifically, we review the results of experiments using an oligonucleotide analog of the decoding region of 16S ribosomal RNA, suggesting that the decoding region is the functional core of the small subunit, that it contacts both mRNA codons and tRNA anticodons, and that it mediates and probably enhances codon-anticodon base pairing, that is, decoding. PMID- 8722006 TI - Pleiotropic effects of mutations at positions 13 and 914 in Escherichia coli 16S ribosomal RNA. AB - Mutations at position 13 or 914 of Escherichia coli 16S ribosomal RNA exert pleiotropic effects on protein synthesis. They interfere with the binding of streptomycin, a translational miscoding drug, to the ribosomes. They increase translational fidelity, and this effect can be related to a perturbation of the higher order structure of the 530 stem-loop, a key region for tRNA selection. In contrast, the structure of the decoding center is not perturbed. The mutations also affect translational initiation, slowing down the formation of the 30S initiation complex. This effect can be related to a destabilization of the pseudoknot helix (17-19/916-918), at the convergence of the three major domains of 16S ribosomal RNA. PMID- 8722007 TI - The pseudouridine residues of ribosomal RNA. AB - Pseudouridine (psi), the most common single modified nucleoside in ribosomal RNA, has been positioned in the small subunit (SSU) and large subunit (LSU) RNAs of a number of representative species. Most of the information has been obtained by application of a rapid primed reverse transcriptase sequencing technique. The locations of these psi residues have been compared. Many sites for psi are the same among species, but others are distinct. In general, the percentage psi in multicellular eukaryotes is greater than in prokaryotes. In LSU RNA, the psi residues are strongly clustered in three domains, all of which are near or connected to the peptidyl transferase center. There is no apparent clustering of psi in SSU RNA. The psi sites in LSU RNA overlap those for the methylated nucleosides, but this is not the case in SSU RNA. There are 265 psi sites known to nucleotide resolution, of which 246 are in defined secondary structures, and 112 of these are in nonidentical structural contexts. All 246 psi sites can be classified into five structural types. Two Escherichia coli psi synthases have been cloned and characterized, one for psi 516 in SSU RNA and one for psi 746 in LSU RNA. The psi 746 synthase recognizes free RNA, but the psi 516 enzyme requires an intermediate RNP particle. Possible functional roles for psi in the ribosome are discussed. PMID- 8722008 TI - Variety of nonsense suppressor phenotypes associated with mutational changes at conserved sites in Escherichia coli ribosomal RNA. AB - To screen for ribosomal RNA mutants defective in peptide chain termination, we have been looking for rRNA mutants that exhibit different patterns of suppression of nonsense mutations and that do not suppress missense mutations at the same positions in the same reporter gene. The rRNA mutations were induced by segment directed randomly mutagenic PCR treatment of a cloned rrnB operon, followed by subcloning of the mutagenesis products and transformation of strains containing different nonsense mutations in the Escherichia coli trpA gene. To date, we have repeatedly obtained only two small sets of mutations, one in the 3' domain of 16S rRNA, at five nucleotides out of the 610 mutagenized (two in helix 34 and three in helix 44), and the other in 23S rRNA at only four neighboring nucleotide positions (in a highly conserved hexanucleotide loop) within the 1.4 kb mutagenized segment. There is variety, however, in the suppression patterns of the mutants, ranging from suppression of UAG or UGA, through suppression of UAG and UGA, but not UAA, to suppression of all three termination codons. The two helices in 16S rRNA have previously been associated both physically and functionally with the decoding center of the ribosome. The 23S region is part of the binding site for the large subunit protein L11 and the antibiotic thiostrepton, both of which have been shown to affect peptide chain termination. Finally, we have demonstrated that the 23S mutant A1093, which suppresses trpA UGA mutations very efficiently, is lethal at temperatures above 36 degrees C (when highly expressed). This lethality is overcome by secondary 23S rRNA mutations in domain V. Our results suggest that specific regions of 16S and 23S rRNA are involved in peptide chain termination, that the lethality of A1093 is caused by high-level UGA suppression, and that intramolecular interaction between domains II and V of 23S rRNA may play a role in peptide chain termination at the UGA stop codon. PMID- 8722009 TI - Structure and evolution of mammalian ribosomal proteins. AB - Mammalian (rat) ribosomes have 80 proteins; the sequence of amino acids in 75 have been determined. What has been learned of the structure of the rat ribosomal proteins is reviewed with particular attention to their evolution and to amino acid sequence motifs. The latter include: clusters of basic or acidic residues; sequence repeats or shared sequences; zinc finger domains; bZIP elements; and nuclear localization signals. The occurrence and the possible significance of phosphorylated residues and of ubiquitin extensions is noted. The characteristics of the mRNAs that encode the proteins are summarized. The relationship of the rat ribosomal proteins to the proteins in ribosomes from humans, yeast, archaebacteria, and Escherichia coli is collated. PMID- 8722010 TI - Location and domain structure of Escherichia coli ribosomal protein L7/L12: site specific cysteine crosslinking and attachment of fluorescent probes. AB - Five different variants of L7/L12 containing single cysteine substitutions, two in the N-terminal (NTD) and three in the C-terminal domain (CTD), were produced, modified with [125I]N-[4-(p-azidosalicylamido)butyl]-3-(2'-pyridyldithio) propionamide ([125I]APDP), a sulfhydryl-specific, heterobifunctional, cleavable photo-cross-linking reagent, and reconstituted into ribosomes. These were irradiated, the total proteins were extracted and reductively cleaved, and the cross-linked proteins were identified. The effect of zero-length disulfide cross linking on binding and activity was also determined. The same sites in L7/L12 were used to attach a rhodamine dye. The formation of ground-state rhodamine dimers caused the appearance of a new absorption band at 518 nm that was used to estimate the extent of interaction of the probes in the free protein and in complexes with L10. The three sites in the CTD, but not the N-terminal sites, cross-linked to L2 and L5 and to 30S proteins S2, S3, S7, S14, and S18 in a manner influenced by elongation factors. Binding to the ribosome and, therefore, function were blocked by zero-length cross-linking within the NTD, but not the CTD. Binding also disrupted rhodamine dimers in the NTD. No rhodamine dimers formed in the CTD. PMID- 8722012 TI - Structure and expression of ribosomal protein genes in Xenopus laevis. AB - In Xenopus laevis, as well as in other vertebrates, ribosomal proteins (r proteins) are coded by a class of genes that share some organizational and structural features. One of these, also common to genes coding for other proteins involved in the translation apparatus synthesis and function, is the presence within their introns of sequences coding for small nucleolar RNAs. Another feature is the presence of common structures, mainly in the regions surrounding the 5' ends, involved in their coregulated expression. This is attained at various regulatory levels: transcriptional, posttranscriptional, and translational. Particular attention is given here to regulation at the translational level, which has been studied during Xenopus oogenesis and embryogenesis and also during nutritional changes of Xenopus cultured cells. This regulation, which responds to the cellular need for new ribosomes, operates by changing the fraction of rp-mRNA (ribosomal protein mRNA) engaged on polysomes. A typical 5' untranslated region characterizing all vertebrate rp-mRNAs analyzed to date is responsible for this translational behaviour: it is always short and starts with an 8-12 nucleotide polypyrimidine tract. This region binds in vitro some proteins that can represent putative trans-acting factors for this translational regulation. PMID- 8722011 TI - Proteins P1, P2, and P0, components of the eukaryotic ribosome stalk. New structural and functional aspects. AB - The eukaryoic ribosomal stalk is thought to consist of the phosphoproteins P1 and P2, which form a complex with protein PO. This complex interacts at the GTPase domain in the large subunit rRNA, overlapping the binding site of the protein L11 like eukaryotic counterpart (Saccharomyces cerevisiae protein L15 and mammalian protein L12). An unusual pool of the dephosphorylated forms of proteins P1 and P2 is detected in eukaryotic cytoplasm, and an exchange between the proteins in the pool and on the ribosome takes place during translation. Quadruply disrupted yeast strains, carrying four inactive acidic protein genes and, therefore, containing ribosomes totally depleted of acidic proteins, are viable but grow with a doubling time threefold higher than wild-type cells. The in vitro translation systems derived from these stains are active but the two-dimensional gel electrophoresis pattern of proteins expressed in vivo and in vitro is partially different. These results indicate that the P1 and P2 proteins are not essential for ribosome activity but are able to affect the translation of some specific mRNAs. Protein PO is analogous to bacterial ribosomal protein L10 but carries an additional carboxyl domain showing a high sequence homology to the acidic proteins P1 and P2, including the terminal peptide DDDMGFGLFD. Successive deletions of the PO carboxyl domain show that removal of the last 21 amino acids from the PO carboxyl domain only slightly affects the ribosome activity in a wild type genetic background; however, the same deletion is lethal in a quadruple disruptant deprived of acidic P1/P2 proteins. Additional deletions affect the interaction of PO with the P1 and P2 proteins and with the rRNA. The experimental data available support the implication of the eukaryotic stalk components in some regulatory process that modulates the ribosomal activity. PMID- 8722013 TI - Structures of prokaryotic ribosomal proteins: implications for RNA binding and evolution. AB - After a long hiatus, the pace of determination of the structures of ribosomal proteins has accelerated dramatically. We discuss here the structures of five ribosomal proteins from Bacillus stearothermophilus: S5, S17, L6, L9, and L14. These structures represent several new motifs. Each of these structures has revealed new insights, and we have developed criteria for recognizing RNA-binding regions of each protein and correlating the structures with such properties as antibiotic resistance. The information here should also prove invaluable in an eventual high-resolution picture of the intact ribosome. PMID- 8722014 TI - Relocation of the unusual VAR1 gene from the mitochondrion to the nucleus. AB - The Var1 protein (Var1p) is an essential, stoichiometric component of the yeast mitochondrial small ribosomal subunit, and it is the only major protein product of the mitochondrial genetic system that is not part of an energy transducing complex of the inner membrane. Interestingly, no mutations have been reported that affect the function of Var1p, presumably because loss of a functional mitochondrial translation system leads to an instability of mtDNA. To study the structure, function and synthesis of Var1p, we have engineered yeast strains for the expression of this protein from a nuclear gene, VAR1U, in which 39 nonstandard mitochondrial codons were converted to the universal code. Immunoblot analysis using an epitope-tagged form of Var1Up showed that the nuclear-encoded protein was expressed and imported into the mitochondria. VAR1U was tested for its ability to complement a mutation in mtDNA, PZ206, which disrupts '3-end processing of the VARI mRNA, causing greatly reduced synthesis of Var1p and a respiratory-deficient phenotype. Respiratory growth was restored in PZ206 mutants by transformation with a centromere plasmid carrying VAR1U under ADH1 promoter control, thus proving that VAR1 function can be relocated from the mitochondrion to the nucleus. Moreover, epitope-tagged Var1Up co-sedimented specifically with small ribosomal subunits in high salt sucrose gradients. The relocation of VAR1 from the mitochondrion to the nucleus provides an excellent system for the molecular genetic analysis of structure-function relationships in the unusual Var1 protein. PMID- 8722015 TI - Structure and function of ribosomal RNA. AB - A refined model has been developed for the folding of 16S rRNA in the 30S subunit, based on additional constraints obtained from new experimental approaches. One set of constraints comes from hydroxyl radical footprinting of each of the individual 30S ribosomal proteins, using free Fe(2+)-EDTA complex. A second approach uses localized hydroxyl radical cleavage from a single Fe2+ tethered to unique positions on the surface of single proteins in the 30S subunit. This has been carried out for one position on the surface of protein S4, two on S17, and three on S5. Nucleotides in 16S rRNA that are essential for P site tRNA binding were identified by a modification interference strategy. Ribosomal subunits were partially inactivated by chemical modification at a low level. Active, partially modified subunits were separated from inactive ones by binding 3'-biotinderivatized tRNA to the 30S subunits and captured with streptavidin beads. Essential bases are those that are unmodified in the active population but modified in the total population. The four essential bases, G926, 2mG966, G1338, and G1401 are a subset of those that are protected from modification by P-site tRNA. They are all located in the cleft of our 30S subunit model. The rRNA neighborhood of the acceptor end of tRNA was probed by hydroxyl radical probing from Fe2+ tethered to the 5' end of tRNA via an EDTA linker. Cleavage was detected in domains IV, V, and VI of 23S rRNA, but not in 5S or 16S rRNA. The sites were all found to be near bases that were protected from modification by the CCA end of tRNA in earlier experiments, except for a set of E site cleavages in domain IV and a set of A-site cleavages in the alpha-sarcin loop of domain VI. In vitro genetics was used to demonstrate a base-pairing interaction between tRNA and 23S rRNA. Mutations were introduced at positions C74 and C75 of tRNA and positions 2252 and 2253 of 23S rRNA. Interaction of the CCA end of tRNA with mutant ribosomes was tested using chemical probing in conjunction with allele-specific primer extension. The interaction occurred only when there was a Watson-Crick pairing relationship between positions 74 of tRNA and 2252 of 23S rRNA. Using a novel chimeric in vitro reconstitution method, it was shown that the peptidyl transferase reaction depends on this same Watson Crick base pair. PMID- 8722016 TI - The elongating ribosome: structural and functional aspects. AB - We determined the positions and arrangements of RNA ligands within the ribosome with a new neutron-scattering technique, the proton-spin contrast-variation. Two tRNAs were bound to the ribosome in the pre-translocational and the post translocational state. The mass centre of gravity of both tRNAs resides at the subunit interface of the body of the 30S subunit. Both tRNAs are separated by an angle of 50-55 degrees, and their mutual arrangement does not change during translocation. The mass centre of gravity moves by 13 +/- 3 A (1A = 0.1 nm) during translocation, corresponding well with the length of one codon. Using an RNase-digestion technique, the length of the mRNA sequence covered by the ribosome was determined to be 39 +/- 3 nucleotides before and after translocation. The ribosome moves like a rigid frame along the mRNA during translocation. In contrast, both tRNAs seem to be located on a movable ribosomal domain, which carries the tRNAs before, during, and after translocation, leaving the microtopography of the tRNAs with the ribosome unaltered. This conclusion was derived from an analysis of the contract patterns of thioated tRNAs on the ribosome. The results have led to a new model of the elongation cycle, which reinterprets the features of the previous "allosteric three-sites model" in a surprisingly simple fashion. Finally, a mutational analysis has identified a single nucleotide of the 23S rRNA essential for the peptidyltransferase activity. PMID- 8722017 TI - Escherichia coli initiator tRNA: structure-function relationships and interactions with the translational machinery. AB - We showed previously that the sequence and (or) structural elements important for specifying the many distinctive properties of Escherichia coli initiator tRNA are clustered in the acceptor stem and in the anticodon stem and loop. This paper briefly describes this and reviews the results of some recently published studies on the mutant initiator tRNAs generated during this work. First, we have studied the effect of overproduction of methionyl-tRNA transformylase (MTF) and initiation factors IF2 and IF3 on activity of mutant initiator tRNAs that are defective at specific steps in the initiation pathway. Overproduction of MTF rescued specifically the activity of mutant tRNAs defective in formylation but not mutants defective in binding to the P site. Overproduction of IF2 increased the activity of all mutant tRNAs having the CUA anticodon but not of mutant tRNA having the GAC anticodon. Overproduction of IF3 had no effect on the activity of any of the mutant tRNAs tested. Second, for functional studies of mutant initiator tRNA in vivo, we used a CAU --> CUA anticodon sequence mutant that can initiate protein synthesis from UAG instead of AUG. In contrast with the wild type initiator tRNA, the mutant initiator tRNA has a 2-methylthio-N6-isopentenyl adenosine (ms2i6A) base modification next to the anticodon. Interestingly, this base modification is now important for activity of the mutant tRNA in initiation. In a miaA strain of E. coli deficient in biosynthesis of ms2i6A, the mutant initiator tRNA is much less active in initiation. The defect is specifically in binding to the ribosomal P site. PMID- 8722019 TI - Peptidyl transferase and beyond. AB - The peptidyl transferase center of the Escherichia coli ribosome encompasses a number of 50S-subunit proteins as well as several specific segments of the 23S rRNA. Although our knowledge of the role that both ribosomal proteins and 23S rRNA play in peptide bond formation has steadily increased, the location, organization, and molecular structure of the peptidyl transferase center remain poorly defined. Over the past 10 years, we have developed a variety of photoaffinity reagents and strategies for investigating the topography of tRNA binding sites on the ribosome. In particular, we have used the photoreactive tRNA probes to delineate ribosomal components in proximity to the 3' end of tRNA at the A, P, and E sites. In this article, we describe recent experiments from our laboratory which focus on the identification of segments of the 23S rRNA at or near the peptidyl transferase center and on the functional role of L27, the 50S subunit protein most frequently labeled from the acceptor end of A- and P-site tRNAs. In addition, we discuss how these results contribute to a better understanding of the structure, organization, and function of the peptidyl transferase center. PMID- 8722018 TI - Identification of ribosome-ligand interactions using cleavage reagents. AB - To characterize ribosome-ligand interactions, we have used a cleavage reagent, 1,10-orthopenanthroline-Cu(II), tethered to various ligands, to cleave nearby regions of rRNA. The phenanthroline is tethered to the ligand using either an internal 4-thiouridine or a terminal thiophosphate. When Cu2+ and a reducing agent, such as mercaptopropionic acid, are present, cleavage of nearby nucleic acids occurs. The cleavage sites can be identified using primer-extension analysis. We have identified rRNA cleavage sites resulting from transcribed tRNAPhe having randomly placed phenanthroline-Cu(II), tRNAPhe with phenanthroline Cu(II) at position 8, and a DNA oligomer complementary to positions 2655-2667 (alpha-sarcin region) with phenanthroline-Cu(II) placed at the 5' end. These results provide important new information on the structure of the rRNA within ribosomal subunits and on the proximity of rRNA neighborhoods to these bound ligands. PMID- 8722020 TI - tRNA-ribosome interactions. AB - Direct measurements of the rates of dissociation of dipeptidyl-tRNA from the ribosome show that hyperaccurate SmP and SmD ribosomes have unstable A-site binding of peptidyl-tRNA, while P-site binding is extremely stable in relation to the wild type. Error-prone Ram ribosomes, on the other hand, have stable A-site and unstable P-site binding of peptidyl-tRNA. At least for these mutant ribosomes, we conclude that stabilization of peptidyl-tRNA in one site destabilizes binding in the other. Elongation factor Tu (EF-Tu) undergoes a dramatic structural transition from its GDP-bound form to its active GTP-bound form, in which it binds aa-tRNA (aminoacyl-tRNA) in ternary complex. The effects of substitution mutations at three sites in domain I of EF-Tu, Gln124, Leu120, and Tyr160, all of which point into the domain I-domain III interface in both the GTP and GDP conformations of EF-Tu, were examined. Mutations at each position cause large reductions in aa-tRNA binding. An attractive possibility is that the mutations alter the domain I-domain III interface such that the switching of EF Tu between different conformations is altered, decreasing the probability of aa tRNA binding. We have previously found that two GTPs are hydrolyzed per peptide bond on EF-Tu, the implication being that two molecules of EF-Tu may interact on the ribosome to catalyze the binding of a single aa-tRNA to the A-site. More recently we found that ribosomes programmed with mRNA constructs other than poly(U), including the sequence AUGUUUACG, invariably use two GTPs per peptide bond in EF-Tu function. Other experiments measuring the protection of aa-tRNA from deacylation or from RNAse A attack show that protection requires two molecules of EF-Tu, suggesting an extended ternary complex. To remove remaining ambiguities in the interpretion of these experiments, we are making direct molecular weight determinations with neutron scattering and sedimentation diffusion techniques. PMID- 8722021 TI - Translational bypassing: a new reading alternative of the genetic code. AB - The translation of the genetic code, once thought to be rigid, has been found to be quite flexible, and several alternatives in its reading have been described. An unusual alternative is translational bypassing, a frameshift event where the transition from frame 0 to another frame occurs by translational bypassing of an extended region of the mRNA sequence rather than by slippage past a single nucleotide, as has been described for most examples of frameshifting. Translational bypassing has been characterized in two cases, T4 gene 60 coding for a topoisomerase subunit and in a trpR-lac'Z fusion. The latter was discovered in our laboratory, and the unique bypass mechanism is investigated further in this study. Using a trpR+1-lac'Z fusion system, we show that the Gln codon at the beginning of lacZ end at the 3' side of the gap is required for bypassing to occur. The Gln codon is part of an mRNA segment that can (potentially) base pair with a segment at the 5' and of Escherichia coli 16S rRNA. A model of trpR+1 lac'Z bypassing is suggested in which the untranslated region of the mRNA is looped out through base pairing between a segment in the 5' end of the 16S rRNA and two sites in the mRNA. Translational bypassing is a newly discovered mechanism of gene expression, and trpR is the first cellular gene identified in which such a mechanism could operate. The understanding of this mechanism and its associated signals may be considered a paradigm for the expression of other genes by this alternative reading of the genetic code. PMID- 8722022 TI - Comparison of functional peptide encoded in the Escherichia coli 23S rRNA with other peptides involved in cis-regulation of translation. AB - A new approach for studying functional rRNA fragments has been developed based on using a plasmid library expressing random fragments of rRNA. A 34 nucleotide long fragment of Escherichia coli 23S rRNA has been identified that renders cells resistant to erythromycin, when expressed in vivo. The rRNA fragment contains a five codon long open reading frame, initiating at GUG and terminating at UAA, with a Shine-Dalgarno sequence located at an appropriate distance from the initiator codon. Translation of this mini-gene is required for the observed erythromycin resistance. Experiments with in vitro translated, or synthetic, peptide indicate the ribosome as a likely target for the action of the identified rRNA-encoded peptide, which apparently remains associated with the ribosome after completion of its translation. The known properties of the rRNA-encoded peptide are compared with information about other functionally active short peptides that can be involved in regulation of translation. PMID- 8722023 TI - Translational regulation of infC operon in Bacillus stearothermophilus. AB - A Bacillus stearothermophilus in vitro translational system has been developed to study the expression of the three cistrons (infC, rpml, and rplT) constituting the infC operon of this bacterium. When directed by homologous in vitro transcribed infC tricistronic mRNA, this system, which consists of partially purified and purified components of the B. stearothermophilus translational apparatus, synthesizes with high efficiency and specificity the three gene products (IF3, L35, and L20) in a ratio similar to that found in vivo (i.e., about 1:6:6). The three cistrons are translationally coupled and expressed in a specific temporal order: a low level of IF3 synthesis stimulates the expression of L35 which, in turn, greatly stimulates the synthesis of L20 and IF3. Protein L20 and an excess of IF3 were found to act as translational feedback inhibitors of the entire operon. The synthesis of IF3 displayed a strong dependence on IF2. This dependence as well as the repressibility by excess IF3 were found to be due to the presence of the rare AUU initiation triplet at the beginning of infC. PMID- 8722024 TI - Termination of translation in eukaryotes. AB - Termination of translation is governed in ribosomes by polypeptide chain release factors (pRF and eRF in prokaryotes and eukaryotes, respectively). In prokaryotes, three pRF have been indentified and sequenced, while in eukaryotes, only a single eRF has been identified to date. Recently, we have characterized a highly conserved protein family called eRF1. At least, human and Xenopus laevis proteins from this family are active as eRFs in the in vitro assay with any of the three stop codons. No structural similarity has been revealed between any of the three pRFs and eRF1 family. Furthermore, GTP-binding motifs have not been revealed, although translation termination in eukaryotes is a GTP-dependent process. We have demonstrated that in eukaryotes a second eRF exists in addition to eRF1, called eRF3. The eRF3 family has two features in common: presence of GTP binding motifs and high conservation of the C-terminal domain structure. The C terminal domain of the X. laevis eRF3 has no RF activity although it stimulates the eRF1 activity considerably at low concentration of the stop codons, conferring GTP dependence to the termination reaction. Without eRF3, the eRF1 activity is entirely GTP independent. Some features of X. laevis eRF3 (C-terminal domain) resemble those of pRF3. The newly identified eRF1 and eRF3 are structurally conserved and distinct from the respective pRF1/2 and pRF3 proteins, pointing to the possibility of different evolution of translation termination machinery in prokaryotes and eukaryotes. Bipartition of the translation termination apparatus probably provides high rate and accuracy of translation termination. PMID- 8722025 TI - Histidine 229 in protein L2 is apparently essential for 50S peptidyl transferase activity. AB - It has recently been suggested that peptidyl transferase activity is primarily a property of ribosomal RNA and that ribosomal proteins may act only as scaffolding. On the other hand, evidence from both photoaffinity labeling studies and reconstitution studies suggest that protein L2 may be functionally important for peptidyl transferase. In the work reported here, we reconstitute 50S subunits in which the H229Q variant of L2 replaces L2, with all other ribosomal components remaining unchanged, and determine the catalytic and structural properties of the reconstituted subunits. We observe that mutation of the highly conserved His 229 to Gin results in a complete loss of peptidyl transferase activity in the reconstituted 50S subunit. This is strong evidence for the direct involvement of L2 in ribosomal peptidyl transferase activity. Control experiments show that, though lacking peptidyl transferase activity, 50S subunits reconstituted with H229Q-L2 appear to be identical with 50S subunits reconstituted with wild-type L2 with respect to protein composition and 70S formation in the presence of added 30S subunits. Furthermore, as shown by chemical footprinting analysis, H229Q-L2 appears to bind 23S RNA in the same manner as wild-type L2. Thus, the effect of H229 mutation appears to be confined to an effect on peptidyl transferase activity, providing the most direct evidence for protein involvement in this function to date. PMID- 8722026 TI - Translational termination efficiency in both bacteria and mammals is regulated by the base following the stop codon. AB - The translational stop signal and polypeptide release factor (RF) complexed with Escherichia coli ribosomes have been shown to be in close physical contact by site-directed photochemical cross-linking experiments. The RF has a protease sensitive site in a highly conserved exposed loop that is proposed to interact with the peptidyltransferase center of the ribosome. Loss of peptidyl-tRNA hydrolysis activity and enhanced codon-ribosome binding by the cleaved RF is consistent with a model whereby the RF spans the decoding and peptidyltransferase centers of the ribosome with domains of the RF linked by conformational coupling. The cross-link between the stop signal and RF at the ribosomal decoding site is influenced by the base following the termination codon. This base determines the efficiency with which the stop signal is decoded by the RF in both mammalian and bacterial systems in vivo. The wide range of efficiencies correlates with the frequency with which the signals occur at natural termination sites, with rarely used weak signals often found at recoding sites and strong signals found in highly expressed genes. Stop signals are found at some recoding sites in viruses where -1 frame-shifting occurs, but the generally accepted mechanism of simultaneous slippage from the A and P sites does not explain their presence here. The HIV-1 gag-pol-1 frame shifting site has been used to show that stop signals significantly influence frame-shifting efficiency on prokaryotic ribosomes by a RF-mediated mechanism. These data can be explained by an E/P site simultaneous slippage mechanism whereby the stop codon actually enters the ribosomal A site and can influence the event. PMID- 8722027 TI - Regulation of the Escherichia coli S10 ribosomal protein operon by heterologous L4 ribosomal proteins. AB - We have cloned the L4 ribosomal protein genes from Morganella morganii and Haemophilus influenza. The sequences of these genes were compared with published sequences for Escherichia coli, Yersinia pseudotuberculosis, and Bacillus stearothermophilus. All five of these L4 genes were expressed in E. coli and shown to function as repressors of both transcription and translation of the E. coli S10 operon. Possible implications for regulation of r-protein synthesis in species other E. coli are discussed. PMID- 8722028 TI - Regulation of translation termination: conserved structural motifs in bacterial and eukaryotic polypeptide release factors. AB - Translation termination requires codon-dependent polypeptide release factors. The mechanism of stop codon recognition by release factors is unknown and holds considerable interest since it entails protein-RNA recognition rather than the well-understood mRNA-tRNA interaction in codon-anticodon pairing. Bacteria have two codon-specific release factors and our picture of prokaryotic translation is changing because a third factor, which stimulates the other two, has now been found. Moreover, a highly conserved eukaryotic protein family possessing properties of polypeptide release factor has now been sought. This review summarizes our current understanding of the structural and functional organization of release factors as well as our recent findings of highly conserved structural motifs in bacterial and eukaryotic polypeptide release factors. PMID- 8722029 TI - Upstream stimulators for recoding. AB - Recent progress in elucidation of 5' stimulatory elements for translational recoding is reviewed. A 5' Shine-Dalgarno sequence increases both +1 and -1 frameshift efficiency in several genes; examples cited include the E. coli prfB gene encoding release factor 2 and the dnaX gene encoding the gamma and tau subunits of DNA polymerase III holoenzyme. The spacing between the Shine-Dalgarno sequence and the shift site is critical in both the +1 and -1 frameshift cassettes; however, the optimal spacing is quite different in the two cases. A frameshift in a mammalian chromosomal gene, ornithine decarboxylase antizyme, has recently been reported; 5' sequences have been shown to be vital for this frameshift event. Escherichia coli bacteriophage T4 gene 60 encodes a subunit of its type II DNA topoisomerase. The mature gene 60 mRNA contains an internal 50 nucleotide region that appears to be bypassed during translation. A 16 amino acid domain of the nascent peptide is necessary for this bypass to occur. PMID- 8722030 TI - A pseudoknot is required for efficient translational initiation and regulation of the Escherichia coli rpsO gene coding for ribosomal protein S15. AB - Escherichia coli ribosomal protein S15 down regulates its own synthesis by binding to its mRNA in a region overlapping the ribosome binding site, called the translational operator. This binding stabilizes a pseudoknot structure that exists in equilibrium with two stem-loop structures. When synthesized in excess over 16S rRNA, S15 binds to its translational operator and traps the ribosome on its loading site in a transient state, preventing the formation of the active ternary (30S-mRNA-rRNA(f)Met) complex. This inhibition can be suppressed by 16S rRNA, which displaces S15 from the mRNA. An extensive mutational analysis showed that the pseudoknot is the structural element required for S15 recognition and in vivo translational control. Specific sequence determinants are located in limited regions of the structure formed by the pseudoknot. An unexpected result is that the pseudoknot can exist in a variety of topologically equivalent structures recognizable and shapable by S15. Based on footprinting experiments and computer graphic modelling, S15 shields the two stems of the pseudoknot, sitting in the major groove of the coaxial stack. PMID- 8722031 TI - The accuracy center of a eukaryotic ribosome. AB - Mutations in yeast ribosomal proteins and ribosomal RNAs have been shown to affect translational fidelity. These mutations include: proteins homologous to Escherichia coli's S4, S5, and S12; a eukaryote specific ribosomal protein; yeast ribosomal rRNA alterations at positions corresponding to 517, 912, and 1054 in 16S E. coli rRNA and to 2658 in the sarcin-ricin domain of 23S E. coli rRNA. Overall there appears to be a remarkable conservation of the accuracy center throughout evolution. PMID- 8722032 TI - Fungal ribotoxins: a family of naturally engineered targeted toxins? AB - alpha-Sarcin, mitogillin, and restrictocin are small (approximately 17 kDa) basic robosome-inactivating proteins (RIPs) produced by the Aspergilli that catalytically inactivate the large ribosomal subunits of all organisms tested to date. These three fungal ribotoxins act as specific ribonucleases by hydrolyzing one single phosphodiester bond in the universally conserved alpha-sarcin domain of 23-28S rRNAs and are among the most potent inhibitors of protein synthesis known. Previous molecular studies of ribotoxins indicated that they belong to the superfamily of ribonucleases and analysis of the mitogillin gene employing PCR mediated site-specific mutagenesis suggests that certain domains in ribotoxins, which share homologies with motifs in ribosome-related proteins, may be responsible for the targeting of ribotoxins to the ribosome. The applications of the ribotoxins as tools in research and their uses as therapeutic and diagnostic agents are also reviewed in this paper. PMID- 8722033 TI - Fragmentation of the ribosome to investigate RNA-ligand interactions. AB - RNA molecules perform a variety of important and diverse functions and, therefore, an understanding of their structure and interaction with proteins and ligands is essential. Large RNA molecules (for example, the ribosomal RNAs) are complex and hence reports describing their fragmentation into functional subdomains has provided a means for their detailed analysis. We present here an in vivo approach to study RNA-ligand interactions. This is based on the concept that an RNA fragment could mimic a drug-binding site present on the intact molecule. Overexpression of the fragment would sequester the drug thereby permitting the continued functioning of the ribosome and, thus, ensuring cell viability. Accordingly, a fragment of 16S rRNA encompassing the spectinomycin binding domain in helix 34 (nucleotides 1046-1065 and 1191-1211) was cloned and in vivo expression resulted in drug resistance. Furthermore, an RNA fragment lacking flanking sequences to helix 34 was also selected from among a pool of random rRNA fragments and shown to confer spectinomycin resistance. A similar in vitro approach is also described for the analysis of rRNA molecules that interact with the yeast elongation factor 3 (EF-3). PMID- 8722034 TI - Antibiotic resistance mechanisms of mutant EF-Tu species in Escherichia coli. AB - Analysis of antibiotic-resistant EF-Tu mutants has revealed a connection between resistance and structural elements that participate in the GTPase switching mechanism. Both random and site-directed mutagenesis methods have yielded sets of purified mutant EF-Tu resistant to kirromycin (kirT) or pulvomycin (pulT). All kirT mutations cluster in the interface of domain 1 and 3 of EF-Tu in its GTP bound conformation, not in that of EF-Tu.GDP. Other evidence also suggests that kirromycin binds to the interface of wild-type EF-Tu, thereby jamming the GTPase switch. Various functional studies reveal two subsequent resistance mechanisms. The first hinders kirromycin binding to EF-Tu.GTP and the second occurs after GTP hydrolysis by rejection of bound kirromycin. All pulT mutations cluster in the three-domain junction interface of EF-Tu. GTP (which is an open hole in EF Tu.GDP) and destabilize a salt-bridge network. Pulvomycin may bind nearby and overlap with tRNA binding. Mutations show that a D99-R230 salt bridge is not essential for the transduction of the GTPase switch signal from domain 1. In vivo and in vitro studies reveal that pulvomycin sensitivity is dominant over resistance. This demands a revision of the current view of the mechanism of pulvomycin inhibition of protein synthesis and may support a translation model with two EF-Tus on the ribosome. Several mutant EF-Tu species display altered behaviour towards aminoacyl-tRNA with interesting effects on translational accuracy. KirT EF-Tu(A375T) is able to reverse the streptomycin-dependent phenotype of a ribosomal protein S12 mutant strain to streptomycin sensitivity. PMID- 8722035 TI - Recognition determinants for proteins and antibiotics within 23S rRNA. AB - Ribosomal RNAs fold into phylogenetically conserved secondary and tertiary structures that determine their function in protein synthesis. We have investigated Escherichia coli 23S rRNA to identify structural elements that interact with antibiotic and protein ligands. Using a combination of molecular genetic and biochemical probing techniques, we have concentrated on regions of the rRNA that are connected with specific functions. These are located in different domains within the 23S rRNA and include the ribosomal GTPase-associated center in domain II, which contains the binding sites for r-proteins L10.(L12)4 and L11 and is inhibited by interaction with the antibiotic thiostrepton. The peptidyltransferase center within domain V is inhibited by macrolide, lincosamide, and streptogramin B antibiotics, which interact with the rRNA around nucleotide A2058. Drug resistance is conferred by mutations here and by modification of A2058 by ErmE methyltransferase. ErmE recognizes a conserved motif displayed in the primary and secondary structure of the peptidyl transferase loop. Within domain VI of rRNA, the alpha-sarcin stem-loop is associated with elongation factor binding and is the target site for ribotoxins including the N-glycosidase ribosome-inactivating proteins ricin and pokeweed antiviral protein (PAP). The orientations of the 23S rRNA domains are constrained by tetiary interactions, including a pseudoknot in domain II and long-range base pairings in the center of the molecule that bring domains II and V closer together. The phenotypic effects of mutations in these regions have been investigated by expressing 23S rRNA from plasmids. Allele-specific priming sites have been introduced close to these structures in the rRNA to enable us to study the molecular events there. PMID- 8722036 TI - Structural and functional implications in the eubacterial ribosome as revealed by protein-rRNA and antibiotic contact sites. AB - Contact sites between protein and rRNA in 30S and 50S ribosomal subunits of Escherichia coli and Bacillus stearothermophilus were investigated at the molecular level using UV and 2-iminothiolane as cross-linkers. Thirteen ribosomal proteins (S3, S4, S7, S14, S17, L2, L4, L6, L14, L27, L28, L29, and L36) from these organisms were cross-linked in direct contact with the RNAs, and the peptide stretches as well as amino acids involved were identified. Further, the binding sites of puromycin and spiramycin were established at the peptide level in several proteins that were found to constitute the antibiotic-binding sites. Peptide stretches of puromycin binding were identified from proteins S7, S14, S18, L18, AND L29; those of spiramycin attachment were derived from proteins S12, S14, L17, L18, L27, and L35. Comparison of the RNA-peptide contact sites with the peptides identified for antibiotic binding and with those altered in antibiotic resistant mutants clearly showed identical peptide areas to be involved and, hence, demonstrated the functional importance of these peptides. Further evidence for a functional implication of ribosomal proteins in the translational process came from complementation experiments in which protein L2 from Halobacterium marismortui was incorporated into the E. coli ribosomes that were active. The incorporated protein was present in 50S subunits and 70S particles, in disomes, and in higher polysomes. These results clearly demonstrate the functional implication of protein L2 in protein biosynthesis. Incorporation studies with a mutant of HmaL2 with a replacement of histidine-229 by glycine completely abolished the functional activity of the ribosome. Accordingly, protein L2 with histidine-229 is a crucial element of the translational machinery. PMID- 8722037 TI - Cotranslational folding of nascent proteins on Escherichia coli ribosomes. AB - Evidence is presented for cotranslational folding of rhodanese or ricin during its synthesis on Escherichia coli ribosomes. During transcription-translation, full-length but enzymatically inactive polypeptides accumulated as peptidyl-tRNA on the ribosomes. These polypeptides were activated and released by subsequent incubation with the bacterial chaperones and with release factor (RF-2). Coumarin was incorporated cotranslationally at the N-terminus of the nascent protein from fluorophore-S-Ac-Met-tRNAf. Changes in fluorescence indicated that DnaJ bound to the nascent proteins and to a fluorescently labeled synthetic peptide corresponding to the N-terminal 17 amino acids of bovine rhodanese. This peptide also bound to 70S ribosomes or 50S subunits but not to 30S subunits. It inhibited activation and RF-2-dependent release of the full-length ribosome-bound rhodanese. A deletion mutant of rhodanese lacking the N-terminal 23 amino acids was not accumulated on the ribosome but was synthesized very efficiently. However, the protein that was formed was enzymatically inactive. DnaJ did not bind to this deletion mutant on ribosomes. We conclude that the chaperone mediated reactions facilitate binding of the N-terminal sequence of nascent proteins to a specific site on 50S ribosomal subunits where it blocks release. The ribosome-bound protein undergoes chaperone-mediated reactions that are required for folding into an enzymatically active conformation. PMID- 8722038 TI - Crystallographic studies of elongation factor G. AB - The elongation factors G (EF-G) and Tu (EF-Tu) go through a number of conformation states in their functional cycles. Since they both are GTPases, have similar G domains and domains II, and have similar interactions with the nucleotides, then GTP hydrolysis must occur in similar ways. The crystal structures of two conformational states are known for EF-G and three are known for EF-Tu. The conformations of EF-G.GDP and EF-Tu.GTP are closely related. EF-Tu goes through a large conformational change upon GTP cleavage. This conformational change is to a large extent due to an altered interaction between the G domain and domains II and III. A number of kirromycin-resistant mutations are situated at the interface between domains I and III. The interface between the G domain and domain V in EF-G corresponds with this dynamic interface in EF-Tu. The contact area in EF-G is small and dominated by interactions between charged amino acids, which are part of a system that is observed to undergo conformational changes. Furthermore, a number of fusidic acid resistant mutants have been identified in this area. All of this evidence makes it likely that EF-G undergoes a large conformational change in its functional cycle. If the structures and conformational states of the elongation factors are related to a scheme in which the ribosome oscillates between two conformations, the pretranslocational and posttranslocational states, a model is arrived at in which EF-Tu drives the reaction in one direction and EF-G in the opposite. This may lead to the consequence that the GTP state of one factor is similar to the GDP state of the other. At the GTP hydrolysis state, the structures of the factors will be close to superimposable. PMID- 8722039 TI - Cotranslational folding of proteins. AB - Many unfolded polypeptides are capable of refolding into their native structure upon the removal of the denaturant. However, the folding of the mature protein during renaturation does not accurately reflect the folding process of nascent proteins in the interior of the cell. This view resulted from the discovery of molecular chaperones known to modulate protein folding. Recent publications discussing the possible role and mechanisms of chaperone action suggest that folding in vivo may be a posttranslational process. Here we discuss data that indicate the final native structure and biological activity can be attainted can be nascent protein on the ribosome, thus supporting the cotranslational folding hypothesis. PMID- 8722040 TI - Elongation factor Tu, a GTPase triggered by codon recognition on the ribosome: mechanism and GTP consumption. AB - The mechanism of elongation factor Tu (EF-Tu) catalyzed aminoacyl-tRNA (aa-tRNA) binding to the A site of the ribosome was studied. Two types of complexes of EF Tu with GTP and aa-tRNA, EF-Tu.GTP-aa-tRNA (ternary) and (EF-Tu.GTP)2.aa-tRNA (quinternary), can be formed in vitro depending on the conditions. On interaction with the ribosomal A site, generally only one molecule of GTP is hydrolysed per aa-tRNA bound and peptide bond formed. The second GTP molecule from the quinternary complex is hydrolyzed only during translation of an oligo(U) tract in the presence of EF-G. The first step in the interaction between the ribosome and the ternary complex is the codon-independent formation of an initial complex. In the absence of codon recognition, the aa-tRNA-EF-Tu complex does not enter further steps of A site binding and remains in the initial binding state. Despite the rapid formation of the initial complex, the rate constant of GTP hydrolysis in the noncognate complex is four orders of magnitude lower compared with the cognate complex. This, together with the results of time-resolved fluorescence measurements, suggests that codon recognition by the ternary complex on the ribosome initiates a series of structural rearrangements that result in a conformational change of EF-Tu, presumably involving the effector region, which, in turn, triggers GTP hydrolysis and the subsequent steps of A site binding. PMID- 8722042 TI - Antigen-specific suppressor factor: missing pieces in the puzzle. AB - Few areas of immunologic research have endured such strident criticism or engendered such fainthearted support as the study of antigen-specific suppression of the immune response. Although enjoying a modest resurgence as a means of promoting or maintaining peripheral tolerance to autoantigens, the study of antigen-specific suppression is not mainstream immunology. The field of immune regulation has, in fact, shifted focus toward explaining the data in terms of the Th1/Th2 paradigm. Indeed, the term suppression has been coopted, by those willing to use it, to describe the bioactivity of conventional cytokines, such as IL-4, IL-10 or TGF beta, which can be inhibitory in certain experimental models. In a very real sense, those who performed much of the early work in the field bear responsibility for the outcast status of suppression. With the increasing number of soluble mediators and cascades of interacting T cells, which populated reviews of the subject in the 1980s, the concept of antigen-specific suppression and suppressor factors simply became too complicated and was dismissed as artifact. Several laboratories have in the past few years made significant advances in the molecular characterization of antigen-specific TsF. Their work, as well as that of our own laboratory have established certain minimal molecular requirements for the expression of TsF bioactivity. PMID- 8722041 TI - The selectins: insights into selectin-induced intracellular signaling in leukocytes. AB - Characteristic features of the inflammatory and immune responses involve the recruitment of leukocytes to sites of tissue injury and the recirculation of lymphocytes through hematopoietic and lymphoid tissues. Recent studies indicate that the regulated cell surface expression of a family of protein adhesion molecules known as selectins and their counterreceptors on both leukocytes and endothelium play critical roles in both biologic processes. Initially, the function of these molecules was thought to be restricted to regulating cell-cell adhesive interactions. Selectin-dependent cell-cell binding has been shown to be essential in localizing leukocytes within tissues by promoting cell rolling along endothelium prior to the development of tight adhesion and subsequent cell migration. However, recent studies suggest that these molecules also play an active role in regulating additional leukocyte functions. This article will review the emerging evidence that indicates a broader and significant role of selectin molecules and their counterreceptors in the initiation of intracellular signaling pathways and regulation of other leukocyte functional responses including degranulation, cytokine expression, activation of the respiratory burst, and T lymphocyte activation. PMID- 8722043 TI - Induction of Th2 cell tolerance to a soluble antigen by blockade of the LFA-1 dependent pathway prevents allergic inflammation. AB - In this article, we show that induction of Th2 cell tolerance prevents antigen induced eosinophil recruitment into the tissue and IgE antibody production, and that ICAM-1/LFA-1 interaction is involved as a costimulatory signal in inducing T cell tolerance to a soluble antigen. In vivo pretreatment with anti-ICAM-1 monoclonal antibody (mAb), anti-LFA-1 mAb, and a soluble antigen inhibited antigen-induced eosinophil recruitment into the airways and IgE antibody production in mice in an antigen-specific manner. In vitro antigen-induced IL-2, IL-4 and IL-5 production were decreased in spleen cells of the mice pretreated with the two mAbs and the antigen, indicating the induction of both Th1 and Th2 cell tolerance in vivo. These results suggest that the induction of antigen specific Th2 cell tolerance by allergen immunotherapy with blockade of the ICAM /LFA-1 interaction would be a rational therapeutic approach to allergic inflammation such as asthma. PMID- 8722047 TI - Pregnancy outcomes in women without gestational diabetes mellitus related to the maternal glucose level. Is there a continuum of risk? AB - OBJECTIVE: To examine selected pregnancy outcomes in women without gestational diabetes mellitus to see whether there was a continuum of risk related to the maternal glucose level. RESEARCH DESIGN AND METHODS: Consecutive women attending two prenatal clinics and three obstetricians in private practice were tested for GDM at the beginning of the third trimester using a 75-g glucose load in the fasting state. The rate of induction, the number of assisted deliveries, the presence of pregnancy-induced hypertension, fetal birth weights, and morbidity were examined with respect to the maternal 2-h glucose level. RESULTS: Data were available for 1,441 women with a 2-h glucose level < 8.0 mmol/l (144 mg/dl). For each 1.0 mmol/l (18 mg/dl) increase in the glucose level, the odds in favor of an assisted delivery increased by 15.2%, and the odds in favor of the baby being admitted to a special care nursery (SCN) increased by 22.6%. There was no significant association between maternal glucose levels and the probability of either pregnancy-induced hypertension or a large-for-gestational-age (LGA) baby after adjustment for other variables. CONCLUSIONS: In normal women there is a continuum of risk related to the maternal glucose level 2 h after a glucose tolerance test for the probability of having an assisted delivery and the likelihood of the baby being admitted to an SCN. The chance of having pregnancy induced hypertension or a LGA baby also increased as the maternal glucose level increased but could be largely explained by an increasing body mass index. PMID- 8722044 TI - Regulatory mechanisms of antitumor T cell responses in the tumor-bearing state. AB - Tumor-bearing hosts develop antitumor immune responses. However, a number of immunosuppressive mechanisms come into operation with the progression of tumor growth. This article will review the observations regarding the modulation of antitumor immune responses in the tumor-bearing state, and consider the mechanisms underlying tumor-induced immune defects, especially in the light of the induction of an abnormal cytokine network. We will also describe the restoration of suppressed antitumor immune responses by administration of a particular cytokine, interleukin-12. PMID- 8722046 TI - Skin-homing T cells in human cutaneous allergic inflammation. AB - The cutaneous lymphocyte-associated antigen (CLA) is a carbohydrate epitope present on memory/effector T cells that infiltrate inflamed skin. E-selectin is the ligand for CLA and is induced under inflammation on endothelial cells. CLA was originally postulated as a phenotype marker for skin-associated T cells. We studied the specific in vitro response to skin-associated allergens of CLA+ and CLA-CD45RO+ T cells in atopic dermatitis (AD) and contact dermatitis (CD), which represent two well-characterized T cell-mediated cutaneous allergic inflammations. Whereas CLA+ T cells from AD patients preferentially responded to house dust mite (HDM) and CLA+ T cells from nickel CD patients showed an increased response to nickel, CLA-T cells showed very little response in both cases. In contrast, tetanus toxoid, a systemically acting antigen, induced a proliferative response in both CLA+ and CLA- cells. Interestingly the response to HDM in patients with asthma +/- AD was preferentially found in the CLA- subset indicating the involvement of different homing receptors for mucosal tissues. Moreover, CLA+ T cells showed enhanced migration through activated human umbilical vein endothelial cell monolayers compared to CLA- T cells. The CLA binding to E-selectin is initially responsible for the extravasation that also involves VLA-4/VCAM-1 and LFA-1/ICAM-1 interactions. We have recently identified IL-8 as an endothelial cell-derived chemokine and the IL-8 receptor type B which control CLA+ T cell migration. Such a CLA-mediated migration would localize memory/effector T cells that respond to antigens and reach the body through inflamed skin. Our data support the existence of a regionalization of the immune system and in particular of the skin immune system. It may allow an efficient distribution of the immune defense to different sites of the body. PMID- 8722045 TI - Moloney leukemia virus-induced cell surface antigen mimicry by monoclonal antibodies. AB - We have investigated antigen-independent modulation of immune responses by monoclonal antibodies directed against both viral and nonviral antigens. BALB/c mice were immunized with monoclonal IgM (i.e. Ab1) specific for either Moloney murine leukemia virus-induced cell surface antigen (MCSA) or the hapten 2,4 dinitrophenyl (DNP). Injection with either Ab1 activated a functional idiotypic (Id) network as evidenced by production of both anti-Id (Ab2) antibodies and anti anti-Id (Ab3) antibodies. A subset of induced Ab3 (designated Ab1'), exhibited specificity for antigen (virus or DNP). In mice immunized with anti-Id antibodies (Ab2), production of Ab3 and Ab1' was also observed. In the MCSA system, antibody induced Ab1' responses were effective in protecting mice from tumor development upon subsequent challenge with live virus. Furthermore, antigen-independent modulation of immunity to both viral and nonviral antigens was found to be thymus dependent. Similar findings in other viral systems suggest that antibody-induced activation of Id networks may prove a viable alternative vaccine strategy that can elicit antigen-specific responses, and in some cases protection, in the apparent absence of exposure to antigen. PMID- 8722048 TI - Long-term glycemic control relates to mortality in type II diabetes. AB - OBJECTIVE: To study the influence of long-term glycemic control on mortality in a cohort of newly detected type II diabetic individuals. RESEARCH DESIGN AND METHODS: A total of 411 newly detected type II diabetic individuals diagnosed between 1972 and 1987 were followed until 31 December 1989. Clinical data concerning fasting blood glucose (FBG) values, body mass index (BMI), type of treatment, and concomitant diseases were collected during 8 randomly selected years. Long-term glycemic control was measured as annual averages of FBG values during these years. Mortality data were obtained from official registers. RESULTS: There were 161 diabetic subjects who died during a mean follow-up time of 7.4 years. In univariate analyses, higher age at diagnosis; higher baseline or average FBG; and the presence of heart disease, cerebrovascular disease, or kidney disease at the beginning or during the course of diabetes were related to higher mortality. Type of diabetes treatment or having a diagnosis of hypertension was not related to mortality. In multiple logistic regression analyses, age at diagnosis and average FBG were independently related to all cause (P = 0.0002), cardiovascular (P = 0.0006), and ischemic heart disease mortality (P = 0.03). No correlations between glycemic control and noncardiovascular deaths were found. In a Cox's regression analysis, average FBG was significantly related to length of survival when age at diabetes diagnosis was taken into account (P < 0.05). Diabetic subjects with average FBG > or = 7.8 mmol/l had 50% higher mortality compared with diabetic subjects with average FBG < 7.8 mmol/l. CONCLUSIONS: Diabetic patients with good long-term glycemic control had a better survival rate than subjects with high average FBG values. The findings should be interpreted cautiously because possible confounding factors such as dyslipoproteinemia and smoking were not studied. PMID- 8722049 TI - Impaired retinal artery blood flow in IDDM patients before clinical manifestations of diabetic retinopathy. AB - OBJECTIVE: To determine whether hemodynamic changes in retinal arteries precede clinical manifestations of diabetic retinopathy and to examine the effects of control of hyperglycemia on retinal artery blood flow. RESEARCH DESIGN AND METHODS: We assessed blood flow in bilateral central retinal arteries in 50 insulin-dependent diabetes mellitus (IDDM) patients without retinopathy and 20 sex- and age-matched control subjects using duplex Doppler sonography. We determined the peak systolic velocity (PSV), end-diastolic velocity (EDV), time averaged velocity (TAV), resistance index (RI), and pulsatility index (PI). RESULTS: PSV, EDV, and TAV were significantly lower in IDDM patients than in control subjects (P < 0.05, P < 0.01, and P < 0.01, respectively). The RI was significantly higher in IDDM patients than in control subjects (P < 0.01) and was significantly correlated with plasma levels of glucose in IDDM patients (r = 0.0.310, P = 0.0248). Multiple regression analysis identified the plasma levels of glucose as a significant determination of RI in IDDM patients. After 14 days of intensive insulin therapy in 7 IDDM patients, the RI and plasma levels of glucose showed significant decreases (P = 0.018, P = 0.001, respectively). CONCLUSIONS: Our results showed that changes in retinal hemodynamics were present before the clinical detection of overt diabetic retinopathy and suggest that the presence of short-term hyperglycemia partly contributes to impaired retinal circulation. PMID- 8722050 TI - Maternal serum triglyceride, glucose tolerance, and neonatal birth weight ratio in pregnancy. AB - OBJECTIVE: To determine the value of measuring serum triglyceride (TG) levels early in pregnancy for predicting late-gestation glucose tolerance and neonatal birth weight ratio (BWR) (birth weight corrected for gestational age). RESEARCH DESIGN AND METHODS: The relationships between morning nonfasting TG measured early in pregnancy (gestational age 12 +/- 6 weeks [mean +/- SD]) and glucose tolerance measured by a 3-h 50-g oral glucose tolerance test (OGTT) late in pregnancy (gestational age 30 +/- 3 weeks) and BWR were investigated in 388 women attending routine antenatal care. The data were analyzed for all women in addition to subgroups of Australian/Western European-born (n = 246) and Asian born (n = 97) women. RESULTS: Morning nonfasting TG positively correlated with the OGTT glucose area under the curve (OGTT-GAUC) (r = 0.23, P < 0.0001) in all subjects. This correlation was stronger in the subset of subjects who had TG measured between 9 and 12 weeks of gestation (r = 0.35, P = 0.0001) and was particularly strong in Asian-born women who had TG measured within this period (r = 0.71, P < 0.0001). Mean TG and the 2- and 3-h OGTT values were higher in Asian born subjects compared with Australian/Western European-born subjects (P = 0.004, P < 0.0001, and P = 0.02, respectively). TG correlated positively with BWR in all subjects (r = 0.12, P = 0.02), in Asian-born subjects (r = 0.23, P = 0.02), and in subjects with gestational diabetes mellitus (GDM) (r = 0.60, P = < 0.001). CONCLUSIONS: TG, if measured between 9 and 12 weeks of gestation, has moderate predictive value for subsequent glucose tolerance in pregnancy. TG is also predictive of BWR in GDM subjects. Further studies are warranted to investigate the role of early TG measurement in the screening and management of GDM. Metabolic heterogeneity exists between Asian-born and Australian/Western European born women, the significance of which is still unclear and warrants further study. PMID- 8722052 TI - Morbidity and mortality in the Wolfram syndrome. AB - OBJECTIVE: To determine the major causes of morbidity and mortality in the autosomal recessive Wolfram syndrome, which is defined by diabetes and bilateral progressive optic atrophy with onset in childhood or adolescence. RESEARCH DESIGN AND METHODS: We abstracted and reviewed the medical records of 68 confirmed cases of Wolfram syndrome identified through a nationwide survey of endocrinologists, ophthalmologists, institutes, and homes for the blind. We also reviewed all available autopsy records. RESULTS: The most common causes of morbidity and mortality were the neurological manifestations of this syndrome and the complications of urinary tract atony. There was a lower frequency of diabetic ketoacidosis, no histologically proven diabetic glomerulosclerosis, and less severe, more slowly progressive, diabetic retinopathy than in classic type I diabetic patients. Mortality in Wolfram syndrome is much higher than in type I diabetes; 60% of Wolfram syndrome patients die by age 35. Recognition of these clinical differences from classic type I diabetes is important for the proper management of Wolfram syndrome patients. CONCLUSIONS: Identification of Wolfram syndrome patients among all diabetic patients presenting in childhood or adolescence is important because the management of patients with this syndrome is different from that of patients with classic type I diabetes. PMID- 8722051 TI - GAD antibodies in NIDDM. Ten-year follow-up from the diagnosis. AB - OBJECTIVE: To study the frequency of antibodies to glutamic acid decarboxylase (GAD) and islet cell antibodies (ICAs) and their predictive value with respect to the development of insulin deficiency in 133 newly diagnosed middle-aged patients with non-insulin-dependent diabetes mellitus (NIDDM) and in 126 control subjects and to study the persistence of GAD antibodies in diabetic patients during the follow-up. RESEARCH DESIGN AND METHODS: The study participants consisted of a well-characterized group of 133 middle-aged newly diagnosed patients with NIDDM and 126 control subjects. The follow-up examinations were performed 5 and 10 years after the baseline. The development of absolute and relative insulin deficiency was based on a stimulated C-peptide level that was undetectable or < 0.70 nmol/l, respectively. GAD antibodies were measured retrospectively from stored samples. RESULTS: The overall prevalence of GAD antibody and ICA positivity at the time of diagnosis was 9.0 and 3.8% in diabetic patients and 1.6 and 0% in the control population, respectively. During the 10-year follow-up, 3 (2.3%) and 10 (7.5%) of the diabetic patients developed absolute and relative insulin deficiency, respectively. Of these, two (67%) and six (60%) had been GAD antibody-positive at the time of diagnosis. The sensitivity and specificity of the GAD antibody to predict absolute or relative insulin deficiency were 67 vs. 94% and 60 vs. 95%, while corresponding figures for ICA were 33 vs. 97% and 20 vs. 98%, respectively. The negative predictive value of GAD antibody testing was higher than positive predictive value (97 vs. 50%). During the follow-up, low grade GAD antibody positivity showed an evanescent nature, whereas the high levels were quite persistent. CONCLUSIONS: In an unselected population of newly diagnosed NIDDM patients, the prevalence of latent autoimmune diabetes in adults was < 10%. While GAD antibody and ICA measured at the time of diagnosis of NIDDM are equally specific predictors of subsequent insulin dependency, the GAD antibody may have a higher sensitivity. Therefore, measurements of GAD antibody may aid the clinician in the choice of treatment of these patients. PMID- 8722053 TI - Progression of retinopathy after change of treatment from oral antihyperglycemic agents to insulin in patients with NIDDM. AB - OBJECTIVE: In insulin-dependent diabetes mellitus, institution of good glycemic control has been shown to retard development of retinopathy even though temporary progression has occurred. Few data have been available from patients with non insulin-dependent diabetes mellitus (NIDDM). To determine the impact of improved glycemic control on retinopathy in patients with NIDDM, we examined, in a case control study, the progression of retinopathy in 94 patients who changed treatment from oral antihyperglycemic agents to insulin. RESEARCH DESIGN AND METHODS: We used the Wisconsin retinopathy scale and related progression of retinopathy during a 2-year observation period to changes in HbA1c after institution of insulin therapy. RESULTS: Progression of retinopathy > or = 3 levels occurred in 23% of the patients and was significantly more common in the patient group in which HbA1c was lowered > or = 3% compared with progression in the group in which HbA1c was lowered < 3% (P = 0.0001; relative risk 3.2; 95% confidence interval 1.5-6.9). CONCLUSIONS: Improved glycemic control as achieved by insulin therapy may be associated with worsening of retinopathy in patients with NIDDM. PMID- 8722054 TI - Fibrinolysis and diabetic retinopathy in NIDDM. AB - OBJECTIVE: By contributing to a prothrombotic state, increased levels of plasminogen activator inhibitor-1 (PAI-1) may be involved in the pathogenesis of the vascular complications of non-insulin-dependent diabetes mellitus (NIDDM). The objective of this study was to compare levels of components of the fibrinolytic system in NIDDM subjects with and without retinopathy. RESEARCH DESIGN AND METHODS: A total of 135 Caucasian NIDDM subjects treated with oral therapy or diet alone were classified by the presence or absence of retinopathy, and fasting blood samples were taken for assay of PAI-1 antigen and activity, tissue plasminogen activator (t-PA), t-PA complexed with PAI-1, euglobulin clot lysis time (a measure of overall fibrinolytic activity), glucose, HbA1c, cholesterol, triglyceride, and insulin levels. RESULTS: Subjects with retinopathy had a longer disease duration than those without (9 vs. 5 years, P < 0.0001) and had lower levels of PAI-1 (PAI-1 antigen) (geometric mean and 95% confidence interval), 13.9 (10.4-18.6) vs. 24.1 (21.2-27.4) ng/ml (P < 0.0005); t-PA antigen, 9.6 (8.6-10.7) vs. 11.5 (10.8-12.3) ng/ml (P < 0.01); t-PA--PAI-1 complexes, 6.2 (5.2-7.2) vs. 7.8 (6.8-8.8) ng/ml (P < 0.05); and insulin, 11.5 (9.3-14.3) vs. 19.5 (17.0-22.3) mU/l (P < 0.0005). Euglobulin clot lysis time was shorter in the subjects with retinopathy, 273 (238-312) vs. 327 (303-352) min. When entered into a logistic regression model, disease duration, PAI-1 antigen, and HbA1c remained as significant independent associates of the presence of retinopathy. CONCLUSIONS: These results do not support the hypothesis that impaired fibrinolysis due to elevated PAI-1 is associated with the development of retinopathy because fibrinolysis is indeed enhanced and PAI-1 lower in subjects with retinopathy. PMID- 8722055 TI - Effect of glipizide treatment on response to an infused glucose load in patients with NIDDM. AB - OBJECTIVE: This study was initiated to compare the effect of sulfonylurea treatment on the response to an infused glucose load of patients with non-insulin dependent diabetes mellitus (NIDDM) at a basal insulin concentration and in response to physiological hyperinsulinemia. RESEARCH DESIGN AND METHODS: We used the insulin suppression test, in which subjects were infused for 180 min with somatostatin, exogenous insulin, and glucose. Since similar steady-state plasma insulin (SSPI) concentrations are reached in all subjects, the resultant steady state plasma glucose (SSPG) concentration permits comparison of the ability of a given individual to maintain glucose homeostasis in response to the infused glucose load. RESULTS: We studied 15 nonobese patients at two different SSPI concentrations, before and after glipizide treatment, at basal (68 +/- 4 pmol/l) and high (470 +/- 31 pmol/l) levels. Values for SSPG concentrations were lower after treatment at both the basal (15.3 +/- 0.5 vs. 18.5 +/- 0.6 mmol/l; P < 0.001) and the high (10.6 +/- 0.7 vs. 14.2 +/- 0.7 mmol/l; P < 0.001) SSPI concentrations. To compare the responses of each patient before and after treatment, we calculated the fractional glucose metabolic rate, i.e., (glucose infusion rate--urinary glucose loss) divided by SSPG. To provide an alternative method of comparing the effect of sulfonylurea treatment, we divided the incremental increase in fractional metabolic glucose rate between the studies done at the low and high SSPI by the incremental increase in SSPI between the two studies (insulin sensitivity index [SI]). CONCLUSIONS: The results of these calculations indicated that glipizide treatment was associated with a significant increase in fractional glucose metabolic rate at a basal insulin concentration (29 +/- 3 to 42 +/- 2 ml.m-2.min-1, P < 0.001), and in response to the incremental change in SSPI (14 +/- 4 to 23 +/- 3 ml.m-2.min-1, P < 0.02). Finally, SI also increased in association with sulfonylurea (0.24 +/- 0.06 to 0.43 +/- 0.07 ml.m-2.min-1/microU.ml-1, P < 0.001). PMID- 8722056 TI - Infected puncture wounds in diabetic and nondiabetic adults. AB - OBJECTIVE: To evaluate bone and soft tissue pathogens resulting from puncture wounds among diabetic and nondiabetic adults. RESEARCH DESIGN AND METHODS: We used a case-control design to compare bacterial pathogens in diabetic and nondiabetic subjects with foot infections precipitated by puncture injuries. We used ICD-9-CM code E920.8 to identify 77 diabetic and 69 nondiabetic patients admitted to the hospital for infected puncture wounds. We identified surgical bone and soft tissue cultures and number and type of organisms per culture. RESULTS: Nondiabetic subjects had significantly less osteomyelitis (13 vs. 35%, P < 0.01) than diabetic subjects and were infected by fewer organisms. Pseudomonas was the most common cause of osteomyelitis among nondiabetic subjects (P < 0.001). Staphylococcus aureus was more common in diabetic bone (P < 0.001) and soft tissue (P < 0.001) infections. Polymicrobial osteomyelitis was more common in diabetic subjects. There was a longer delay until diabetic subjects received medical treatment compared with nondiabetic subjects (8.7 vs. 5.3 days, P < 0.002). Diabetic subjects were more likely to have neuropathy (P < 0.001) and to have sustained their injuries while barefoot (P < 0.006). CONCLUSIONS: Puncture wounds in diabetic subjects were commonly associated with polymicrobial infections. Pseudomonas was the most common cause of nondiabetic osteomyelitis. These results have implications for differential emergent and chronic treatment of puncture wounds in diabetic versus nondiabetic subjects. PMID- 8722057 TI - Biomedical and psychiatric risk factors for retinopathy among children with IDDM. AB - OBJECTIVE: Illness duration and glycemic control influence the development of retinopathy in childhood-onset insulin-dependent diabetes mellitus (IDDM). Psychiatric disorders and sociodemographic factors also affect diabetes-related outcomes. However, biomedical and psychosocial factors have not been examined together in modeling the risk of retinopathy. RESEARCH DESIGN AND METHODS: We conducted a single-site prospective longitudinal study of 66 children (aged 8-13 years) newly diagnosed with IDDM. Repeated assessments served to derive psychiatric diagnoses. Poor glycemic control was defined as the upper 15th percentile of all HbA1 values. After a median follow-up of 10 years, severity of retinopathy was determined. It was modeled with a stepwise polychotomous regression procedure using antecedent biomedical and psychosocial variables. RESULTS: Young adults with childhood-onset IDDM were found to be at increased risk of retinopathy the longer they had IDDM, the more persistently they evidenced poor antecedent glycemic control, and the longer they suffered from depressive illness. These three factors operated individually and additively, with duration of IDDM conferring a baseline level of risk. In depressed patients (27%), depression onset antedated the detection of retinopathy generally by 7 years. CONCLUSIONS: Duration of childhood-onset IDDM confers a baseline level of risk of retinopathy irrespective of glycemic control; antecedent clinical depression is also a risk factor. Depression therefore may serve as a marker of vulnerability and help to identify a subgroup of patients at risk for complications. The findings raise the question whether timely treatment of depression could forestall diabetic retinopathy. PMID- 8722058 TI - Steadily high IDDM incidence over 4 years in Sardinia. AB - OBJECTIVE: To verify whether the high incidence of insulin-dependent diabetes mellitus (IDDM) in Sardinia is an epidemic outbreak or a steady phenomenon. RESEARCH DESIGN AND METHODS: All newly diagnosed cases of IDDM with onset in patients 0-29 years of age between 1 January 1989 and 31 December 1992 among residents in Sardinia were obtained from the Sardinian IDDM Incidence Registry. The local IDDM patient association (Associazione Diabete Infantile Giovanile) served as the secondary and independent source. RESULTS: The completeness of ascertainment was 91%. The age-standardized mean annual incidence of IDDM (per 100,000) was 34.4 in the 0- to 14-year-old age-group and 26.2 in the entire 0- to 29-year-old range, respectively. Men-to-women ratios were 1.38 and 1.55, respectively. Seasonal variation in incidence was observed, with a peak in fall and winter and a nadir in summer. CONCLUSIONS: Sardinia has a very high and steady IDDM incidence rate, which is up to fivefold that of other Italian regions and Mediterranean countries and approaches the Finnish top rate in the world. Interaction between the genetic peculiarity of Sardinians and still unidentified powerful environmental agents is likely to account for the phenomenon. PMID- 8722059 TI - Exercise testing as a long-term predictor of the development of microalbuminuria in normoalbuminuric IDDM patients. AB - OBJECTIVE: To determine the association between exercise-induced albuminuria and the development of microalbuminuria over 10 years in subjects with insulin dependent diabetes mellitus (IDDM) who were initially normoalbuminuric. RESEARCH DESIGN AND METHODS: Thirty-two patients with IDDM and a resting urinary albumin/creatinine ratio (UA/UC) < 2.1 mg/mmol (< 15 micrograms/min) were exercised after water loading on a treadmill for 20 min at double their resting heart rate. UA/UC was determined before and after exercise. The exercise test was considered positive if the UA/UC was > 4.3 mg/mmol (> 30 micrograms/min). Results were compared with resting UA/UC after a 10-year follow-up. Persistent microalbuminuria was defined as a UA/UC > 2.1 mg/mmol (> 15 micrograms/min) in each of two early-morning urine collections. RESULTS: Five patients developed persistent microalbuminuria after 10 years, and four patients were predicted by a positive exercise test. Two patients with positive exercise tests did not develop persistent microalbuminuria. The sensitivity of the exercise test for the development of microalbuminuria was 80% (95% confidence interval [CI] 65.8-94.2%) and the specificity was 92.9% (95% CI 83.9-100%). The postexercise UA/UC was positively associated with the UA/UC after 10 years (P = 0.005, R2 = 0.31). This association was independent of HbA1, systolic blood pressure, body mass index, and duration of diabetes, but HbA1 remained an independent predictor (P = 0.02) of UA/UC at follow-up. CONCLUSIONS: Exercise testing may be useful for identifying normoalbuminuric IDDM patients who are susceptible to the later development of microalbuminuria. PMID- 8722061 TI - Fructosamine measurements in the adolescent with type I diabetes. PMID- 8722060 TI - Screening for NIDDM in nonpregnant adults. A review of principles, screening tests, and recommendations. AB - The economic burden of diabetes in the U.S. is now more than 90 billion dollars per year (113-115). In the diabetes community, there is a feeling that aggressive measures to detect diabetes in earlier stages may be warranted. Regardless of whether evidence is convincing that there are benefits from early detection of diabetes (3,116,117), screening activities have been sanctioned by many organizations and are now taking place. Operational research is needed to define more clearly the who, when, where, and how of screening and the effectiveness of screening programs (4,118). Diabetes screening programs should be comprehensive and should educate about diabetes and provide appropriate interpretation of both positive and negative screening results. It is essential that programs have diagnostic follow-up and refer those with newly diagnosed diabetes for treatment. In addition, individuals with previously diagnosed diabetes encountered during screening activities should be made aware of the importance of glycemic control and follow-up with their health care providers. PMID- 8722062 TI - Severe hypoglycemia and type I diabetes with isolated ACTH deficiency. PMID- 8722063 TI - American Diabetes Association scientific sessions, 1995. Magnesium deficiency, atherosclerosis, and health care. PMID- 8722064 TI - Adverse events and their association with treatment regimens in the diabetes control and complications trial. AB - OBJECTIVE: To describe the incidence of adverse events associated with intensive versus conventional therapy of insulin-dependent diabetes mellitus (IDDM) as implemented in the Diabetes Control and Complications Trial (DCCT). RESEARCH DESIGN AND METHODS: The DCCT was a randomized, controlled clinical trial conducted at 26 centers in the U.S. and 3 centers in Canada. All data were collected from patient notifications of events and/or standardized, quarterly interviews that were validated and analyzed at a data coordinating center as events per 100 patient-years. The 1,441 volunteers were between the ages of 13 and 39 with IDDM for 1-15 years. Average length of follow-up was 6.5 years (range 3-9). Subjects were randomly assigned to conventional or intensive diabetes treatment. RESULTS: The two treatment groups did not differ in mortality, major morbidity secondary to accidents, or ketoacidosis. However, intensive therapy was associated with a threefold increase in the risk of severe hypoglycemia (for hypoglycemia requiring assistance, the event rate per 100 patient-years was 61.2 in the intensive treatment group versus 18.7 in the conventional treatment group; for hypoglycemia involving coma or seizure, the rate was 16.3 vs. 5.4). Intensive therapy was also associated with a 73% higher risk of becoming overweight. There was a 46% reduction in the incidence of vaginitis in the intensive treatment group, but there were no significant differences in the rates of other infections. CONCLUSIONS: The major adverse effect of intensive therapy of IDDM is a threefold increase in the risk of severe hypoglycemia with potentially serious sequelae. An increased incidence of becoming overweight, the long-term significance of which has yet to be determined, was also observed. Because the results of the DCCT were attained in highly selected, healthy IDDM patients who received attentive clinical management and frequent health education, DCCT adverse event rates may not reflect incidence or prevalence rates that would be expected in nonselected populations or in other clinical settings. PMID- 8722065 TI - Immunogenetic, clinical, and demographic characterization of childhood type I diabetes in New Zealand. AB - OBJECTIVE: To examine the relationship between genetic susceptibility alleles and islet cell antibodies (ICAs) in type I diabetes. RESEARCH DESIGN AND METHODS: The human leukocyte antigen (HLA)-DQB1 alleles and ICA levels of all incident type I diabetic cases in patients < 20 years of age diagnosed in 1990 and 1991 in Canterbury, New Zealand, were determined. RESULTS: The mean annual incidence rate for type I diabetes over the 24 months was 19.0/100,000 patient-years (95% confidence interval [CI] 13.5-26.0/100,000), which was considerably higher than rates observed between 1982 and 1989 (11.7/100,000; 95% CI 9.6-14.3/100,000). ICAs > or = 10 Juvenile Diabetes Foundation units (JDF U) were present in 84.6% of the subjects, but there was a higher prevalence of ICA-negative (ICA-) subjects among those diagnosed during the winter months. The frequencies of the susceptibility allele DQB1*0302 and susceptibility genotype 0302/0201 were 71.8% and 43.5%, respectively. Subjects with 0302 tended to be younger (mean age 8.3 +/ 5.1 years) than those with nonsusceptibility types (mean age 11.8 +/- 4.7 years, P = 0.056). The probability of being ICA positive (ICA+) was not significantly different between subjects with allele 0302 (85.7%) and those without it (81.8%). All seven patients negative for ICA were homozygous for DQB1 nonaspartate-57. There was no clustering of the immunogenetic markers with demographic and clinical characteristics apart from age at diagnosis. CONCLUSIONS: No direct relationship was observed between DQB1-defined genetic susceptibility and ICA at diagnosis, suggesting that variations at the DQB1 locus are not linked to the expression of this autoimmune marker of beta-cell destruction. PMID- 8722066 TI - Albuminuria and 24-h ambulatory blood pressure in normoalbuminuric and microalbuminuric NIDDM patients. A longitudinal study. AB - OBJECTIVE: To assess the long-term relationships between 24-h ambulatory blood pressure (AMBP), urinary albumin excretion (UAE) rate, and metabolic control in non-insulin-dependent diabetes mellitus (NIDDM) patients with normo- and microalbuminuria. RESEARCH DESIGN AND METHODS: We conducted a prospective study of 23 NIDDM patients (11 with normoalbuminuria and 12 with microalbuminuria) receiving standard clinical care, including antihypertensive treatment, attending the outpatient clinic and 8 healthy control subjects. Twenty-four-hour AMBP and UAE were measured synchronously in addition to fasting plasma glucose, HbA1c, and serum creatinine at baseline and after 4.6 (4.2-5.1) years [mean (range)]. RESULTS: Baseline systolic, but not diastolic, 24-h AMBP was significantly higher in diabetic patients compared with control subjects (146/80 [16/11] vs. 133/78 [9/9] mmHg, P < 0.05), but was similar in normoalbuminuric (143/81 [11/11] mmHg) and microalbuminuric (148/80 [20/10] mmHg) patients during strict blood pressure control. The annual increase in 24-h AMBP was equivalent in diabetic patients (0.6/-0.2 [2.6/1.5] mmHg/year) and control subjects (0.7/0.2 [1.2/1.4] mmHg/year, NS) and not significantly different from zero. Overall UAE did not change in control subjects (5.6 [1.6] vs. 4.4 [1.9]) (geometric mean [antilog SD]) or in the normoalbuminuric (8.7 [1.7] vs. 11.3 [3.0] micrograms/min) and microalbuminuric (35.7 [2.1] vs. 34.5 [3.2] micrograms/min) patients. In diabetic patients, the annual change in UAE correlated significantly with the annual change in the systolic (r = 0.61, P < 0.002) and diastolic (r = 0.54, P < 0.008) 24-h AMBP. In microalbuminuric patients, only the annual increase in systolic 24 h AMBP correlated significantly with the annual change in UAE (r = 0.71, P = 0.010), whereas in the normoalbuminuric patients, only the annual increase in diastolic 24-h AMBP and the annual change in UAE were significantly correlated (r = 0.66, P = 0.026). In a stepwise multiple linear regression analysis, the annual progression in albuminuria in NIDDM patients was significantly determined by increases in systolic (parameter estimate 0.018, SE 0.006, P < 0.008) as well as in diastolic 24-h AMBP (parameter estimate 0.026, SE 0.011, P < 0.033). CONCLUSIONS: In an outpatient clinical setting, 24-h AMBP is similar in NIDDM patients with normo- and microalbuminuria. Alterations in both 24-h AMBP and UAE are on average moderate and equivalent compared with those in healthy control subjects. Although the average change in albuminuria is small, a progression in albuminuria relates to increments in both systolic and diastolic 24-h AMBP. PMID- 8722067 TI - The interrelationship between ethnicity and gestational diabetes in fetal macrosomia. AB - OBJECTIVE: To determine the possibility of an ethnic influence on the development of macrosomia (birth weight > 90th percentile for gestational age) in gestational diabetes mellitus (GDM). RESEARCH DESIGN AND METHODS: We prospectively followed all African-American and Latino women enrolled in the Temple diabetes-in pregnancy program. GDM was diagnosed in 103 African-American and 36 Latino women during the study period (1991-1994) according to the criteria of Carpenter and Coustan. All women were treated according to our previously published protocols. Data were collected on gestational weight gain, previous history of macrosomia, body mass index (BMI), and level of maternal glycemic control. RESULTS: Insulin therapy was required in 53 women (37.5%) to maintain fasting blood glucose levels at < 95 mg/dl and 2-h postprandial levels at < 120 mg/dl. Macrosomia developed in 50% of the neonates of Latino women versus 19% of neonates of African-American women (relative risk 2.68; 95% confidence interval 1.57-4.59). Potential confounding factors were not significantly different between the Latino and African-American women: mean blood glucose 96.6 +/- 15.7 vs. 96.5 +/- 22.4 mg/dl; BMI 29.0 +/- 5.5 vs. 31.5 +/- 8.2 kg/m2; pregnancy weight gain 29.2 +/- 12.7 vs. 30.9 +/- 20.5 lb; and parity 1.8 +/- 1.5 vs. 1.6 +/- 1.4, respectively. CONCLUSIONS: We have demonstrated that Latino women with GDM are at higher risk for having macrosomic infants in comparison with African-American women. This ethnic variation in fetal growth may be due to varying influences of in utero growth promoters among these populations as well as underlying genetic factors. PMID- 8722068 TI - Congenital malformations in pregnancies complicated by NIDDM. AB - OBJECTIVE: To determine whether the use of oral hypoglycemic agents during early pregnancy is associated with a risk of congenital malformations in infants of mothers with non-insulin-dependent diabetes mellitus (NIDDM) independent of maternal metabolic control. RESEARCH DESIGN AND METHODS: From a prospectively collected data-base of pregnancies complicated by diabetes at a large urban medical center, we identified 332 consecutive infants born to women with NIDDM who did not participate in a preconceptional diabetes care program. Stepwise logistical regression was used to identify maternal characteristics that were independently associated with risks of major and minor congenital malformations in infants. RESULTS: Overall, 56 (16.9%) of the 332 infants were born with congenital anomalies (11.7% major anomalies and 5.1% minor anomalies). Analysis of data from subgroups of women who were treated with diet therapy, exogenous insulin, or sulfonylurea compounds during the first 8 weeks of gestation did not reveal statistically significant differences in major or minor malformation rates among the three groups. Stepwise logistic regression analysis revealed two maternal characteristics that were independently associated with major malformations in infants: maternal HbA1c at initial presentation for care (direct relationship; P = 0.0007) and the maternal age at onset of diabetes (inverse relationship; P = 0.009). The risk of major malformations was unrelated to the mode of antidiabetic therapy during early pregnancy. No relationship was found between maternal glycemia or treatment modality and rates of minor congenital anomalies. CONCLUSIONS: These data indicate that, in the absence of special preconceptional care, NIDDM is associated with a risk for major congenital anomalies that is in the range reported for pregnancies complicated by insulin dependent diabetes mellitus. Moreover, the risk in individual patients appears to be related to maternal glycemic control rather than to the mode of antidiabetic therapy during early pregnancy. PMID- 8722069 TI - Improved postprandial metabolic control after subcutaneous injection of a short acting insulin analog in IDDM of short duration with residual pancreatic beta cell function. AB - OBJECTIVE: To compare postprandial metabolic control after subcutaneous injection of a short-acting insulin analog [Lys(B289),Pro(B29)] (Lispro) or human regular insulin (Humulin R U-100 [Hum-R]) in insulin-dependent diabetes mellitus (IDDM) of short duration with residual beta-cell function. RESEARCH DESIGN AND METHODS: Six IDDM patients (age 25 +/- 2 years, diabetes duration 14 +/- 2 months, HbA1c 6.4 +/- 0.5%) with residual pancreatic beta-cell function (fasting plasma C peptide 0.19 +/- 0.02 nmol/l) were studied on three different occasions. Postbreakfast plasma glucose was maintained at approximately 7.1 mmol/l by means of intravenous insulin until either 1200 when 0.1 U/kg Hum-R was injected or until 1225 when 0.1 U/kg of either Hum-R or Lispro was injected subcutaneously. Lunch (mixed meal, 692 Kcal) was served at 1230 (0 min). Six nondiabetic control subjects were also studied. RESULTS: After Lispro administration, the 120-min plasma glucose decreased more (6.1 +/- 0.3 mmol/l) than after injection of Hum-R at -30 min (7.7 +/- 0.3 mmol/l) or -5 min (9.9 +/- 0.2 mmol/l). By the end of the study, plasma glucose was still lower after Lispro was injected (6.7 +/- 0.3 mmol/l) than after Hum-R was injected at -30 min (7.6 +/- 0.3 mmol/l) or -5 min (7.3 +/- 0.2 mmol/l) (P < 0.05). Two IDDM patients required glucose to prevent hypoglycemia after being injected with Lispro, but four required glucose after being injected with Hum-R at -5 min (Lispro approximately 27 mmol glucose infused between 90 and 240 min; Hum-R approximately 80 mmol between 240 and 390 min). After Lispro, plasma insulin peaked earlier (at 30 min, 342 +/- 29 pmol/l) than after Hum-R injection at -30 min (at 90 min, 198 +/- 28 pmol/l) and was superimposable on that of nondiabetic subjects. In Hum-R injected at -5 min, plasma insulin peaked later (at 120 min) and subsequently remained greater than in the two other studies. CONCLUSIONS: Despite the lack of a time interval between injection and meal, Lispro controls postprandial plasma glucose concentration better than Hum-R given 30 min before meals and, to an even greater extent, better than Hum-R given 5 min before meals. In addition, Lispro minimizes the risk of postprandial hypoglycemia, thus closely mimicking the postprandial glucose homeostasis of nondiabetic subjects. IDDM patients with residual pancreatic beta-cell function are the ideal candidates for prandial use of Lispro because they can maintain near-normoglycemia longer after subcutaneous analog injection because of residual endogenous insulin secretion. PMID- 8722070 TI - Increased foot pressures after great toe amputation in diabetes. AB - OBJECTIVE: To compare peak pressures on the sole of the foot in non-insulin dependent diabetic patients with isolated, unilateral amputations of the great toe and first metatarsal with the patients' contralateral, intact foot. RESEARCH DESIGN AND METHODS: Eleven patients with a unilateral great toe and partial first metatarsal amputation of at least 6 months duration were evaluated with the F Scan in-shoe pressure measurement system. Patients were studied in the same brand and style of footwear--a thin, rubber-soled, canvas boat shoe. We compared mean peak plantar foot pressures under the first metatarsal, lesser metatarsals, lesser toes, and heel in feet with and without a great toe amputation using the Wilcoxon's matched pairs signed-rank test. RESULTS: Peak foot pressures were significantly higher under the first metatarsal head (P = 0.046), lesser metatarsal heads (P < 0.001), and toes (P < 0.001) in feet with a great toe amputation compared with the contralateral foot without an amputation. Pressure under the heel was higher on the contralateral foot (P < 0.01). CONCLUSIONS: After a great toe amputation, pressure distribution of the foot is significantly altered. Because preamputation risk factors such as peripheral neuropathy, foot deformity, and limited joint mobility for many of these patients remain unchanged, an increase in foot pressures contributes to an increased risk of reulceration and reamputation in these patients. PMID- 8722071 TI - Atropine inhibits the increase in gastric emptying during hypoglycemia in humans. AB - OBJECTIVE: To study the effect of a cholinergic muscarinic blockade on the gastric emptying rate during insulin-induced hypoglycemia in healthy subjects. RESEARCH DESIGN AND METHODS: In eight healthy subjects, the rate of gastric emptying of an isotope-labeled meal was assessed by a scintigraphic technique during normoglycemia and hypoglycemia with simultaneous infusion of either atropine or saline. Blood glucose concentrations were controlled by an insulin glucose clamp. RESULTS: The median time for emptying 50% of the liquid phase from the stomach (T50) was 24.9 min (range 13.9-120.0) during normoglycemia compared with 8.1 min (range 3.6-16.5) during hypoglycemia without atropine infusion (P = 0.0005). The T50 for the solid phase was 26.8 min (range 9.7-74.0) and 43.1 min (range 29-57.8), respectively (P = 0.007). During hypoglycemia with atropine infusion, T50 was 40.7 min (range 10.0-120.0) for the liquid phase and 111.4 min (range 38.9-120.0) for the solid phase, not statistically different from normoglycemic examinations. CONCLUSIONS: Cholinergic muscarinic blockade with atropine inhibits the increase in gastric emptying during hypoglycemia. Vagal activity seems to be an important determinant of gastric emptying during hypoglycemia. PMID- 8722072 TI - Resource utilization and costs of care in the diabetes control and complications trial. AB - OBJECTIVE: To describe in detail the resources used and costs incurred in the clinical management of patients with insulin-dependent diabetes mellitus (IDDM) in the Diabetes Control and Complications Trial (DCCT). RESEARCH DESIGN AND METHODS: The resources used for intensive and conventional therapy and to deal with the side effects of therapy were assessed at each of the 29 DCCT clinics and summarized. Unit costs were derived from the DCCT, manufacturers, and Medicare and chosen to reflect what an item would cost to a single-payer national health system. Costs were calculated as the product of resources used and unit costs. The costs of the research component of the DCCT were not included. RESULTS: In the DCCT, the annual cost of intensive therapy ($4,000 and $5,800/year for multiple daily injections and continuous subcutaneous insulin infusion, respectively) was approximately three times the cost of conventional therapy ($1,700/year). A large portion of the difference in cost was related to the greater frequency of outpatient visits and the greater resources used in self care. CONCLUSIONS: DCCT intensive therapy is more expensive than conventional therapy, but it offers the hope of cost savings as a result of averted complications. PMID- 8722073 TI - Risk factors for poor glycemic control in diabetic children in France. AB - OBJECTIVE: To test whether immigrant versus nonimmigrant origin, among other factors, is associated with differences in glycemic control in diabetic children. RESEARCH DESIGN AND METHODS: A cross-sectional multicenter survey was performed in 165 French diabetic children aged 7-13 years and their parents. HbA1c was measured by high-performance liquid chromatography in a single laboratory (reference value: 5.1 +/- 0.6%). Standardized questionnaires were used to evaluate cognitive, familial, and socioeconomic factors and adherence to diabetes regimen. RESULTS: The mother's place of birth was found to account for 6.6% of the HbA1c variance. Two groups of children were identified with French-speaking immigrant (n = 27) and native French mothers (n = 138) who had significantly different HbA1c values of 9.1 +/- 8% vs. 8.1 +/- 1.4%, respectively (P < 0.001). Adherence to diabetes treatment was significantly lower in immigrant mothers for diet and insulin changes (P < 0.01), as well as their level of diabetes knowledge (P < 0.01). This immigrant group had no significant differences in parents' socioeconomic status, educational background, family status, family functioning, diabetes education background, and social support compared with the native French mothers' group. CONCLUSIONS: Despite no differences in socioeconomic status, a group of children with less well-controlled diabetes was identified by the mother's place of birth. Cultural factors have a crucial importance in glycemic control in diabetic children and in parents' adherence to treatment and must be taken into account in the development of educational programs. PMID- 8722074 TI - Transcomplementation of HLA DQA1-DQB1 in DR3/DR4 and DR3/DR9 heterozygotes and IDDM in Taiwanese families. AB - OBJECTIVE: To study the human leukocyte antigen (HLA)-DQ heterodimers in the susceptible DR haplotypes for patients with insulin-dependent diabetes mellitus (IDDM) in Taiwan. RESEARCH DESIGN AND METHODS: Extended class II HLA haplotypes were studied in 57 unrelated IDDM patients, 31 simplex IDDM families, and 105 unrelated control subjects recruited from the same area in Taiwan. Class II HLA genotyping was based on PCR-SSO DNA typing techniques. Extended class II HLA haplotypes were deduced unequivocally by the Taiwanese pedigree studies. RESULTS: DR3/DR3, DR3/DR4, and DR3/DR9 genotypes were strongly associated with IDDM susceptibility in this population. In addition to the reported DR3/DR4 in Caucasians, the heterozygotic effect of DR3/DR9 for IDDM was remarkable in the Taiwanese population. Extended HLA haplotypes studies revealed that DRB1*0301/DQA1*0501/DQB1*0201, DRB1*0405/DQA1*0301/DQB1*0302, and DRB1*0405/DQA1*0301/DQB1*0401 were the susceptible haplotypes in this population. There were several hypothetical ways to produce susceptible HLA-DQ heterodimers to explain the susceptibility carried by DR3/DR4 and DR3/DR9 genotypes. Among all DR4 subtypes, only DRB1*0405 was associated with the increased risk of IDDM. CONCLUSIONS: These data strongly suggest that the HLA-DR-associated IDDM susceptibility is most likely explained by the formation of the susceptible DQ heterodimers encoded by the DQA1/DQB1 either in cis or in trans. PMID- 8722075 TI - Cyclosporin A does not delay insulin dependency in asymptomatic IDDM patients. AB - OBJECTIVE: To measure the effects of cyclosporin A (CyA) with no insulin therapy on glucose tolerance and beta-cell function in the preclinical phase of insulin dependent diabetes mellitus (IDDM). RESEARCH DESIGN AND METHODS: beta-cell responses to the intravenous glucose tolerance test (IVGTT), hyperglycemic clamp, intravenous arginine, and intravenous glucagon were evaluated before and after a 6-month course of CyA in seven patients (mean age 19.6 years) with asymptomatic IDDM. RESULTS: Initial insulin secretory responses were severely decreased when the patients were compared with eight healthy control subjects: IVGTT (1 + 3 min): 106 +/- 16 vs. 884 +/- 190 pmol/l (P < 0.001); hyperglycemic clamp: 102 +/- 16 vs. 310 +/- 42 pmol/l (P < 0.001); intravenous arginine: 346 +/- 72 vs. 1104 +/- 168 pmol/l (P < 0.01); and intravenous glucagon: 170 +/- 37 vs. 247 +/- 35 pmol/l (NS). The beta-cell responses remained markedly abnormal after 6 months of CyA, although the response to intravenous glucose and oral glucose tolerance tests improved in three subjects. All the patients became insulin-dependent after 5-36 months. CONCLUSIONS: CyA alone is not a suitable treatment for asymptomatic IDDM. Earlier identification of subjects with substantial beta-cell secretory capacity and newer nontoxic intervention strategies are required for the prevention of IDDM. PMID- 8722076 TI - Nutrition principles and diabetes. A role for "lente carbohydrate"? AB - The current nutrition recommendations of the American Diabetes Association (ADA) represent a thoughtful synthesis of much current data. They depart from tradition by not advocating specific figures for total fat and carbohydrate intake. Rather, since many issues are still topics of scientific debate, they endorse the principle of individualization and set guidelines accordingly. One topic that may be worthy of further debate is the principle of "spreading the nutrient load," or lengthening the absorption time. This principle covers the effects of altered meal frequency, viscous dietary fibers, low-glycemic index foods, and inhibitors of carbohydrate absorption. In its simplest form it is illustrated by studies of altered meal frequency ("nibbling versus gorging"). Reducing the size and increasing the frequency of meals has been shown acutely to result in lower mean blood glucose and insulin levels over the day in type II diabetes and to result in reduced 24-h urinary C-peptide losses. In the longer term in nondiabetic subjects, total and low-density lipoprotein cholesterol levels are reduced, together with fasting apolipoprotein B and serum uric acid levels, as additional risk factors for coronary heart disease. These and other physiological effects make slowing carbohydrate absorption ("lente carbohydrate") a potentially useful therapeutic modality. However, of the possible ways of slowing absorption, only alteration in meal frequency was of general interest in the current ADA nutrition recommendations. Nevertheless, the effects of slowing carbohydrate absorption by various means may have beneficial metabolic effects in diabetes and may support the use of ethnic foods in diets compatible with further modifications identified more favorably in the current nutrition recommendations (e.g., increased use of monounsaturated fat). PMID- 8722077 TI - Is it time to draw the curtain on immune intervention trials in newly diagnosed patients with IDDM? PMID- 8722078 TI - Albuminuria is an independent predictor of carotid intima-media thickness and atherosclerosis in NIDDM patients. PMID- 8722080 TI - Size of the pancreas in type I diabetic children and adolescents. PMID- 8722079 TI - Eating attitude and behavior in IDDM patients. PMID- 8722082 TI - Intrauterine devices are safe and effective contraceptives for type I diabetic women. PMID- 8722081 TI - The possible role of insulin release in cardiovascular morbidity and mortality in NIDDM patients. PMID- 8722083 TI - Insulin and proinsulin secretion in subjects with abnormal glucose tolerance and a mitochondrial tRNA(Leu(UUR)) mutation. PMID- 8722084 TI - The pharmacological treatment of hyperglycemia in NIDDM. American Diabetes Association. PMID- 8722085 TI - Purification of acid phosphatase I from germinating seeds of Vigna sinensis. AB - Acid phosphatase I (AP-I) is the major isoform of Vigna acid phosphatase. It is constitutively expressed in seed cotyledons during germination. AP-I was separated from other isoforms and purified to homogeneity by three simple purification steps; (NH4)2SO4 precipitation, and phosphocellulose and DEAE cellulose column chromatography. The activity of AP-I was not affected by 1 mM Mg2+, Mn2+, Ca2+, Co2+ or Pb2+, but severely inhibited by 1 mM Cu2+, Fe3+, Hg2+, Mo6+ or Zn2+. AP-I has both phosphatase and pyrophosphatase activities, and is highly stable even at 50 degrees. PMID- 8722086 TI - Partial purification and characterization of a calcium-dependent protein kinase in rice leaves. AB - A protein from rice leaves, which was partially purified by sequential chromatography on DE52, MONO-Q and Superose 12, presented calcium-dependent protein kinase (CDPK) activity. This protein kinase phosphorylated the substrate, histone III-S, in a Ca(2+)-dependent manner and the half-maximum concentration of Ca2+ to protein kinase activity (EC50) was 1 microM. This phosphorylation was independent of phosphatidylserine and a phorbol ester. The apparent M(r) of the protein kinase, as determined by phosphorylation in SDS-polyacrylamide gel containing histone III-S, was 45 k. This kinase was found to react differently from other protein kinases, such as protein kinase C from rat brain or CDPK from soybean leaves, owing to the absence of a phospholipid or phorbol ester dependency. CDPK phosphorylated three endogenous proteins as detected by in vitro phosphorylation on two-dimensional PAGE. PMID- 8722087 TI - 6-O-alpha-L-Arabinopyranosyl-beta-D-glucopyranosides as aroma precursors from passion fruit. AB - The 6-O-alpha-L-Arabinopyranosyl-beta-D-glucopyranosides of linalool, benzyl alcohol and 3-methyl-but-2-en-1-ol were isolated from passion fruit (Passiflora edulis) by adsorption chromatography on XAD-2 resin, then further extracted on the same resin after partial enzymic hydrolysis and semi-preparative chromatography on RP-18 phase by HPLC. Their structures were identified by 1H NMR spectroscopy and mass spectral analysis and by methylation analysis of the carbohydrate moieties. PMID- 8722088 TI - Extracellular polysaccharides produced by tuberose callus. AB - A high yield of extracellular polysaccharide (ECP) was obtained from callus cultures of tuberose (Polianthes tuberosa), which could be separated into an unadsorbed and two acidic fractions (TPS-1, -2) by ion-exchange column chromatography. The yields of each fraction were markedly increased by the addition of 10(-5) M 2,4-dichlorophenoxyacetic acid to the medium. Of the three fractions, the amount of TPS-1 accounted for over 60% of total yield of ECP, which was a predominant polysaccharide consisting of arabinose (Ara), mannose (Man) and galactose (Gal) as major neutral monosaccharides. Judging from the patterns of electrophoresis and ultra-centrifugation, TPS-1 was identified to be homogeneous. Methylation and GC-mass spectrometry analyses of this fraction revealed the presence of 1,2,3-linked Man, 1-linked Ara, 1,3-linked Ara, 1-linked Gal and 1,3,4-linked glucuronosyl (GlcUA) residues in a molar ratio of 1.0:1.08:0.85:0.75:1.08. Based on additional analyses of the mild acid hydrolysate and the absolute configuration of the constituent monosaccharides, a possible structure for TPS-1 was a glucuronomannan possessing the unit of -->4) beta-D-GlcUAp-(1-->2)-alpha-D-Manp-(1--> with branching at the C-3 position, where -->1)-alpha-L-Araf, -->1)-beta-D-galp, -->1)-alpha-L-Araf-(3-->1)-alpha-L Araf or -->1)-alpha-L-Araf-(3-->1)-beta-D-Galp were attached randomly. About 35% of the GlcUA moieties were present as methyl esters. Further confirmation was made by 1H and 13C NMR spectroscopy. PMID- 8722089 TI - Minor constituents from Bupleurum fruticosum roots. AB - A new triterpenoid saponin, 23-acetoxy,16 alpha-hydroxy-13,28-epoxyolean-11-en-3 beta-yl--beta-D-glucopyranosyl- (1-->2)---beta-D-glucopyranosil(1-->3)-beta-D fucopyranoside, has been isolated from the roots of Bupleurum fruticosum, along with eight coumarins, scopoletin, scoparone, prenyletin, capensin, fraxetin, aesculetin,7-(3-methyl-2-butenyloxy-6-methoxycoumarin, 7-(2-hydroxy-3-methyl-3 butenyloxy)-6-methoxycoumarin and 5,7-dihydroxy-6-methoxy-8-(3-methyl-2 butenyl)coumarin. The last compound has not been previously encountered in nature. The structures were determined by analysis of NMR spectral data including two-dimensional techniques. PMID- 8722090 TI - Phenolic and triterpenoid glycosides from Aster batangensis. AB - A new phenolic glycoside, asterbatanoside A [p-hydroxyacetophenone-4-O-beta-D xylopyranosyl-(1-->6)-beta-D- glucopyranoside], and two new triterpenoid saponins, asterbatanoside B [2 alpha,3 beta,23-trihydroxyolean-12-en-28-oic acid 28-O-beta-D-glucopyranosyl-(1-->6)-beta-D-glucopyranoside] and asterbatanoside C [3-O-beta-D-glucopyranosyl-2 beta,3 beta,23-trihydroxyolean- 12-en-28-oic acid-28 O-beta-D-glucopyranoside] were isolated from the roots of Aster batangensis. Their structures were determined by spectroscopic methods and chemical evidence. The total synthesis of asterbatanoside A is also reported. PMID- 8722091 TI - Acylated delphinidin glycosides in the blue-violet flowers of Consolida armeniaca. AB - Four new acylated delphinidin 3,7-glycosides were isolated from the blue-violet flowers of Consolida armeniaca as major anthocyanin pigments. The first pigment was based on delphinidin 3,7-diglucoside and was identified as delphinidin 3-O-[6 O-(malonyl)-beta-D-glucopyranoside]-7-O- [6-O-(4-O-(6-O-(p-hydroxybenzoyl)-beta-D glucopyranosyl)-p-hydroxybenzoy l)- beta-D-glucopyranoside] by spectral methods. The other three acylated anthocyanins were based on delphinidin 3-glucoside-7 sophoroside. The second pigment was determined to be delphinidin 3-O-[6-O (malonyl)-beta-D-glucopyranoside]-7-O-[2-O-(beta-D- glucopyranosyl)-6-O-(4-O-(6-O (p-hydroxybenzoyl)-beta-D-glucopyranosyl)- p- hydroxybenzoyl)-beta-D glucopyranoside]. The third pigment was elucidated to be delphinidin 3-O-[6-O (malonyl)-beta-D-glucopyranoside]-7-O-[2-O-(6-O-(p- hydroxybenzoyl)-beta-D glucopyranosyl)-6-O-(4-O-(6-O-(p-hydroxybenzoyl)- beta-D- glucopyranosyl)-p hydroxybenzoyl)-beta-D-glucopyranoside], and the fourth as delphinidin 3-O-[6-O (malonyl)-beta-D-glucopyranoside]-7- O-glucopyranosyl[(2-O-(6-O-(p hydroxybenzoyl)-beta-D-glucopyranosyl)-6-O - (4-O-(6-O-(p-hydroxybenzoyl)-beta-D glucopyranosyl)-p-hydroxybenzoyl)-be ta-D- glucopyranoside]. PMID- 8722092 TI - Acylated pelargonidin 3-sambubioside-5-glucosides in Matthiola incana. AB - Ten acylated pelargonidin 3-sambubioside-5-glucosides were isolated from the red purple flowers of Matthiola incana, and also pelargonidin 3-glucoside was isolated from the brownish-red flowers of this plant. FAB mass measurements of 10 acylated anthocyanins gave their molecular ions [M]+ at 903-1195 m/z, which were based on acylated pelargonidin 3-sambubioside-5-glucosides with malonic acid, sinapic acid, p-coumaric acid, caffeic acid and/or ferulic acid. This was confirmed by the analysis of NMR spectra and the experiments of acid and alkaline hydrolysis. By spectral and chemical methods, seven of the 10 pigments were determined to be pelargonidin 3-O-[2-O-(2-O-(acyl-I)-beta-D-xylopyranosyl)- 6-O (acyl-II)-beta-D-glucopyranoside]-5-O-[6-O-(malonyl)-beta-D- glucopyranoside], in which acyl moieties varied between sinapic, ferulic, caffeic and p-coumaric acids. The occurrence of these pigments was examined in 10 red-purple, 10 salmon pink, three apricot and three copper colour cultivars of M. incana by HPLC. The acylated pelargonidin 3-sambubioside-5-glucosides were present as the dominant pigments in the red-purple, salmon-pink and apricot colour cultivars. On the other hand, pelargonidin 3-glucoside was present as a dominant anthocyanin in the copper colour cultivars and also pelargonidin 3-sambubioside-5-glucoside was confirmed by HPLC as a minor pigment in the copper colour flowers. PMID- 8722093 TI - Two isoflavones from the bark of Petalostemon purpureus. AB - Two new isoflavones, 6,7,8,3',4',5'-hexamethoxyisoflavone and 7,8,3',4',5' pentamethoxyisoflavone, have been isolated and characterized from the combined root bark and stem bark of Petalostemon purpureus. PMID- 8722094 TI - Steroid alkaloid glycosides from Solanum coccineum. AB - In addition to solamargine, isoanguivine and solasonine, two new steroid alkaloid glycosides, xylosylsolamargine and xylosyl-beta-solamarine, have been isolated from the aerial parts of Solanum coccineum, the structures of which have been elucidated as (25R)-3 beta-[O-beta-D-xylopyranosyl-(1-->2)-O-alpha-L rhamnopyranosyl- (1-->4)-O-[alpha-L-rhamnopyranosyl-(1-->2)]-beta-D glucopyranosyloxy+ ++]-22 alpha N-spirosol-5-ene and (25S)-3 beta-[O-beta-D xylopyranosyl-(1-->2)-O-alpha-L-rhamnopyranosyl- (1-->4)-O-[alpha-L rhamnopyranosyl-(1-->2)]-beta-D-glucopyranosyloxy+ ++]-22 beta N-spirosol-5-ene. PMID- 8722095 TI - The molecular basis of pathogenesis of cerebral malaria. PMID- 8722096 TI - Transferrin binding protein two interacts with both the N-lobe and C-lobe of ovotransferrin. AB - The present study was initiated to identify the region(s) of ovotransferrin involved in binding to the bacterial transferrin receptors from Haemophilus paragallinarum and Haemophilus avium. Ovotransferrin was digested with either trypsin or thermolysin to obtain its N-lobe and C-lobe fragments. The individual fragments were then purified by a combination of gel exclusion and ion-exchange chromatography. Solid phase binding experiments with the individual fragments demonstrated that the C-lobe fragments blocked the binding of horse radish peroxidase-conjugated ovotransferrin to the transferrin receptors and that much higher concentrations of the N-lobe fragment were required for any detectable blocking. Affinity isolation of the bacterial transferrin receptor from the two Haemophilus species revealed that both native ovotransferrin and its C-lobe fragment were capable of isolating two iron repressible outer membrane proteins. These 95 and 60 kDa outer membrane proteins correspond to Tbp1 and Tpb2, respectively. In contrast, the N-lobe fragment was capable of isolating Tbp2 of H. paragallinarum but not that of H. avium. The inability of the N-lobe and C lobe fragments from ovotransferrin and human transferrin to support the growth of iron-limited cultures of H. paragallinarum and Neisseria meningitidis, respectively, suggested that interaction with both lobes is necessary for efficient iron acquisition. PMID- 8722097 TI - Lactic acid is the factor in blood cell extracts which enhances the ability of CMP-NANA to sialylate gonococcal lipopolysaccharide and induce serum resistance. AB - In previous work, a factor which enhances the ability of cytidine 5'-monophospho N-acetyl neuraminic acid (CMP-NANA) to sialylate gonococcal lipopolysaccharide (LPS) was liberated at 4 degrees C in diffusates from high M(r) fractions of blood cell sonicates. The diffusates also contained CMP-NANA and converted serum susceptible gonococci to resistance. The enhancer has now been separated from CMP NANA and material absorbing at 260 nm by HPLC on mu Bondapak-10 NH2. Resistance inducing activity was found only in fractions containing CMP-NANA and recovery was poor (about 25%). However, addition of enhancer fractions to CMP-NANA substantially increased its resistance inducing activity. Blood cell sonicates dialysed at 18-20 degrees C released enhancer in diffusates. These were ultrafiltered (nominal cut off 3000 Da) and fractionated on Biogel P2 which removed saccharides and most material absorbing at 260 nm. Over 90% of a fraction which was enhancer-active in nanogram quantities was identified by nuclear magnetic resonance (NMR) spectroscopy and gas chromatography/mass spectometry (GC/MS) as lactic acid. A fraction with similar properties was obtained from a different batch of diffusate by fractionation on Dowex 1. Authentic lithium L lactate in nanogram quantities enhanced LPS sialyation by CMP-NANA and increased its serum resistance inducing activity. These results have important implications for gonococcal pathogenicity. PMID- 8722098 TI - Isolation and characterization of chromosomal mTn 10 insertion mutations affecting K88 fimbriae production in Escherichia coli. AB - mTn 10 transposon mutagenesis of Escherichia coli producing K88 fimbria was carried out in order to identify host factors involved in the regulation of the fae (K88) operon and the production of K88 fimbriae. Five independent chromosomal insertion mutants were obtained which showed an increased expression of K88 fimbriae. Inverse PCR and nucleotide sequencing were carried out to characterize the mutations. One insertion affected the Ipp gene, encoding the major outer membrane lipoprotein. Another mutation was found to be located in the Irp gene, encoding the 'global' regulatory protein Lrp (leucine responsive regulatory protein). A third mutant was found to affect the expression of rfaF, encoding heptosyltransferase II, which resulted in a partially wild-type and partially Re Rd1 type of LPS. A fourth mutation affected sseB, a gene involved in serine sensitivity of E. coli cells. Another mutant contained an insertion in an unknown region of the E. coli genome. The mutants were further characterized with respect to K88 as well as K99 fimbriae production. PMID- 8722099 TI - Trichomonas vaginalis haemolysis: pH regulates a contact-independent mechanism based on pore-forming proteins. AB - There is a controversy in literature about involvement of secreted factors in the pathogenetic mechanisms of Trichomonas vaginalis, described mostly as contact dependent. We found that the protozoan, under triggering conditions, is able to release molecules that lead to lysis without direct contact between parasite and target cells as a prerequisite. In this paper we characterize contact-independent cytotoxicity using the red blood cell as a cellular model. Contact-independent haemolysis is a phenomenon were pH exerts a key role, triggering the secretion of a lytic molecule and regulating its activity. A partial physicochemical characterization of the haemolytic factor suggests that a protein of M(r) > 30 kDa could be the effector responsible for damage. Furthermore, the parasite induced membrane permeabilization, detected by measuring potassium escape from the target cell, and an effective osmotic protection by carbohydrates allowed us to relate the previously described pore-forming mechanism involved in contact dependent cytotoxicity with the contact-independent lysis. PMID- 8722100 TI - Using a new inbred fish model and cultured fish tissue cells to study Aeromonas hydrophila and Yersinia ruckeri pathogenesis. AB - An inbred strain of the southern platyfish, Xiphophorus maculatus, was used as a host for Aeromonas hydrophila and Yersinia ruckeri infections. The infections were initiated by holding the platyfish in inoculation baths containing dilutions of virulent A. hydrophila or Y. ruckeri strains. Inoculating the platyfish in this manner resulted in a dose-dependent mortality over a range of bacterial input from 10(5) to 10(8) A. hydrophila and 10(6) to 10(8) Y. ruckeri/ml. Clinical manifestations of A. hydrophila infections were noted in infected platyfish that eventually died, but not in platyfish that survived. In this model, the Y. ruckeri infected fish died before obvious signs of infection were detected. The A. hydrophila strain used to establish the infections was recovered from the kidney and intestine of infected fish that died, but not from survivors receiving the same inoculation dose. Both infective bacteria were tested for the ability to invade a number of different fish and human cultured cells. A hydrophila strain TF7 did not invade of the cells tested, whereas the Y. ruckeri strain invaded fish derived cultured cells, but not human derived Hep-2 cells. PMID- 8722102 TI - Spotlight on ECRInet's SELECTplus membership services. PMID- 8722101 TI - Surgical helmet systems. AB - A surgical helmet system (SHS) is either a loose-fitting hood or a hood combined with an integral gown (called a toga system). SHSs are used during orthopedic procedures to decrease the patient's risk of deep wound infection by keeping skin and other particles from the surgeon's face from falling into the open surgical site. They are also used to protect surgical staff from infectious blood splashes and potentially infectious aerosols generated by power tools during orthopedic procedures - and it is this aspect of SHS use that has attracted particular attention, given the intensified interest in healthcare worker (HCW) protection (e.g., against human immunodeficiency virus [HIV], tuberculosis [TB], and hepatitis C virus [HCV]) in hospitals today. Ironically, the very factors that have led to SHSs' recent popularity may limit the need for their use, inasmuch as facilities have instituted so many other effective infection control methods against bloodborne and airborne pathogens. Although SHSs do provide splash protection against bloodborne pathogens, institutions may already by adequately protecting their staff against this hazard. In addition, they may be using SHSs to provide other types of patient and HCW protection that are already being provided by existing infection control measures. SHSs are available in a wide variety of configurations. We evaluated a total of six SHSs: one hood and one toga system each from three manufacturers. The units we tested were among each manufacturer's bestselling systems. We examined how well they would protect patients against deep wound infections and HCWs against splashes and aerosolized particles. We also evaluated their comfort and ease of use for wearers. We found that all of the SHSs provided sufficient patient protection against particles shed by the surgeon. We also found that the hood-only units and the hood components of the toga units provided sufficient HCW splash protection. (The gown portion of the toga systems failed our fluid-resistance criterion. However, we did not test the separate surgical gowns that are used with the hood-only units, and we do not know if they provide any greater level of protection than the toga fabric does). We rated all hood-only units Acceptable. We rated all toga units Acceptable-Not Recommended because the level of patient and HCW protection provided by the toga systems did not justify their significantly higher cost. We did find differences in the level of HCW respiratory protection these systems provided against aerosols. However, we do not believe that this should be a significant selection consideration because there is no established need for respiratory protection during orthopedic procedures. (Respiratory protection is needed against TB aerosols, but hip and knee arthroplasty procedures are usually elective and would be delayed on any active or suspected TB patient until the patient is noninfectious). To establish the context for this study, we first present a Guidance Section, "Do You Need Surgical Helmet Systems for Orthopedic Procedures?" In this section, we discuss the infection risks present during orthopedic procedures. We also outline the protective measures that institutions may already have in place to reduce these risks. We then discuss how SHSs fit into the infection control picture. Supplementary material in this section includes a discussion of the respiratory protection needed against Mycobacterium tuberculosis and a table of factors affecting the choice among SHSs, respirator masks, and surgical masks for patient and HCW protection. Ultimately, the choice to use SHSs may be dictated by the training and preference of the surgical staff. But this Evaluation will help facilities to (1) make this decision and, if appropriate, (2) select the best system for their needs... PMID- 8722103 TI - Errors in using blood glucose monitors in hospitals. PMID- 8722104 TI - Possible cross-contamination with Boehringer Mannheim AccuData GTS glucose test station. PMID- 8722105 TI - Excessive cornstarch powder on latex (Esmark) bandages. PMID- 8722106 TI - Quality control problems with ITC's coagulation reagent tubes. PMID- 8722107 TI - Fish models for environmental carcinogenesis: the rainbow trout. AB - Progress over the past 30 years has revealed many strengths of the rainbow trout as an alternative model for environmental carcinogenesis research. These include low rearing costs, an early life-stage ultrasensitive bioassay, sensitivity to many classes of carcinogen, a well-described tumor pathology, responsiveness to tumor promoters and inhibitors, and a mechanistically informative nonmammalian comparative status. Low-cost husbandry, for example, has permitted statistically challenging tumor study designs with up to 10,000 trout to investigate the quantitative interrelationships among carcinogen dose, anticarcinogen dose, DNA adduct formation, and final tumor outcome. The basic elements of the trout carcinogen bioassay include multiple exposure routes, carcinogen response, husbandry requirements, and pathology. The principal known neoplasms occur in liver (mixed hepatocellular/cholangiocellular adenoma and carcinoma, hepatocellular carcinoma), kidney (nephroblastoma), swim bladder (adenopapilloma), and stomach (adenopapilloma). Trout possess a complex but incompletely characterized array of cytochromes P450, transferases, and other enzymic systems for phase I and phase II procarcinogen metabolism. In general, trout exhibit only limited capacity for DNA repair, especially for removal of bulky DNA adducts. This factor, together with a high capacity for P450 bioactivation and negligible glutathione transferase-mediated detoxication of the epoxide, accounts for the exceptional sensitivity of trout to aflatoxin B1 carcinogenesis. At the gene level, all trout tumors except nephroblastoma exhibit variable and often high incidences of oncogenic Ki-ras gene mutations. Mutations in the trout p53 tumor suppressor gene have yet to be described. There are many aspects of the trout model, especially the lack of complete organ homology, that limit its application as a surrogate for human cancer research. Within these limitations, however, it is apparent that trout and other fish models can serve as highly useful adjuncts to conventional rodent models in the study of environmental carcinogenesis and its modulation. For some problems, fish models can provide wholly unique approaches. PMID- 8722109 TI - Pulmonary reactions to organic dust exposures: development of an animal model. AB - Acute inhalation of organic dusts such as cotton, hay, silage, grain, animal confinement, or compost dust can result in illness characterized by fever, pulmonary inflammation, chest tightness, and airway obstruction. These agricultural materials are complex mixtures of plant, bacterial, and fungal products. Elucidation of the time course of disease onset, the mechanisms of disease progression, and the identity of etiologic agents is essential for effective prevention and treatment. Toward this end, animal models for acute organic dust-induced reactions have been developed and characterized. Information concerning the applicability of various animal models to humans and progress toward elucidation of causative agents and mechanisms of action is presented. PMID- 8722108 TI - Pharmacokinetics and molecular detoxication. AB - This paper presents a comprehensive overview of the pharmacokinetic parameters used from in vivo and in vitro studies that are important in order to understand the major conceptual approaches of toxicokinetics and the disposition of environmental chemicals. In vitro biochemical information concerning the detoxication of environmental chemicals is also presented. The discussion leads to a more complete appreciation for the use of in vitro measurements for in vivo correlations. The concept of interspecies scaling in the interpolation and extrapolation of fundamental biochemical metabolic processes is illustrated with a number of examples. Additional examples of in vitro-in vivo correlations are presented in the evaluation of the impact of chemical exposure to humans. Finally, several important metabolic detoxication enzymes are presented, including the mammalian microsomal cytochrome P450 and flavin-containing monooxygenases as well as carboxylesterases and glucuronosyltransferases, to provide insight into the processes of chemical detoxication in mammalian tissue and blood. Because interspecies scaling and the pharmacokinetics of chemical disposition have already shown their usefulness in understanding some examples of chemical disposition, our summary focuses on showing the usefulness of the pharmacokinetic equations and providing confidence in using the approach for in vitro-in vivo correlations. Ultimately, the presentation may provide the reader with a conceptual framework for future evaluation of the human health risks associated with environmental toxicants. PMID- 8722111 TI - Selected new developments in computational chemistry. AB - Molecular dynamics is a general technique for simulating the time-dependent properties of molecules and their environments. Quantum mechanics, as applied to molecules or clusters of molecules, provides a prescription for predicting properties exactly (in principle). It is reasonable to expect that both will have a profound effect on our understanding of environmental chemistry in the future. In this review, we consider several recent advances and applications in computational chemistry. PMID- 8722110 TI - Biomarker research in neurotoxicology: the role of mechanistic studies to bridge the gap between the laboratory and epidemiological investigations. AB - There is an increasing interest in the development and validation of biomarkers for use in biochemical/molecular epidemiological studies. Though the area of neurotoxicology has received much attention in the past several years, it still lags behind with regard to the development of biomarkers, particularly those of health effects and susceptibility. This review discusses several aspects of biomarker research as it relates to neurotoxic compounds and focuses on selected agents (organophosphorus insecticides, styrene, n-hexane, carbon disulfide, acrylamide), which have been the subject of a number of investigations in animals and humans. While traditional biomonitoring approaches and novel techniques (e.g., hemoglobin adducts) provide several measurements for monitoring exposure to neurotoxic chemicals, potential markers of genetic susceptibility have been seldom investigated in a neurotoxicology context. Furthermore, the complexity of the nervous system, together with the multiplicity of end points and the limited knowledge of the exact mechanism(s) of action of neurotoxicants, has led to only limited advancements in the development of biomarkers for neurotoxic effects. Significant progress in this area will depend upon an increased understanding of the cellular, biochemical, and molecular targets directly involved in neurotoxicity. PMID- 8722112 TI - Toxicology of chlorofluorocarbon replacements. AB - Chlorofluorocarbons (CFCs) are stable in the atmosphere and may reach the stratosphere. They are cleaved by UV-radiation in the stratosphere to yield chlorine radicals, which are thought to interfere with the catalytic cycle of ozone formation and destruction and deplete stratospheric ozone concentrations. Due to potential adverse health effects of ozone depletion, chlorofluorocarbon replacements with much lower or absent ozone depleting potential are developed. The toxicology of these compounds that represent chlorofluorohydrocarbons (HCFCs) or fluorohydrocarbons (HFCs) has been intensively studied. All compounds investigated (1, 1-dichloro-1-fluoroethane [HCFC-141b], 1,1,1,2-tetrafluoroethane [HFC-134a], pentafluoroethane [HFC-125], 1-chloro- 1,2,2,2-tetrafluoroethane [HCFC-124], and 1,1-dichloro-2,2,2-trifluoroethane [HCFC-123]) show only a low potential for skin and eye irritation. Chronic adverse effects on the liver (HCFC 123) and the testes (HCFC-141b and HCFC-134a), including tumor formation, were observed in long-term inhalation studies in rodents using very high concentrations of these CFC replacements. All CFC replacements are, to varying extents, biotransformed in the organism, mainly by cytochrome P450-catalyzed oxidation of C-H bonds. The formed acyl halides are hydrolyzed to give excretable carboxylic acids; halogenated aldehydes that are formed may be further oxidized to halogenated carboxylic acids or reduced to halogenated alcohols, which are excretory metabolites in urine from rodents exposed experimentally to CFC replacements. The chronic toxicity of the CFC replacements studied is unlikely to be of relevance for humans exposed during production and application of CFC replacements. PMID- 8722113 TI - Exposure, metabolism, and toxicity of rare earths and related compounds. AB - For the past three decades, most attention in heavy metal toxicology has been paid to cadmium, mercury, lead, chromium, nickel, vanadium, and tin because these metals widely polluted the environment. However, with the development of new materials in the last decade, the need for toxicological studies on those new materials has been increasing. A group of rare earths (RE) is a good example. Although some RE have been used for superconductors, plastic magnets, and ceramics, few toxicological data are available compared to other heavy metals described above. Because chemical properties of RE are very similar, it is plausible that their binding affinities to biomolecules, metabolism, and toxicity in the living system are also very similar. In this report, we present an overview of the metabolism and health hazards of RE and related compounds, including our recent studies. PMID- 8722114 TI - Pesticides: an important but underused model for the environmental health sciences. AB - Pesticides are high-volume, widely used, environmental chemicals and there is continuous debate concerning their possible role in many chronic human health effects. Because of their known structures, known rates of application, and the presence of a large occupationally exposed population, they are not only important in their own right but are ideal models for the effects of environmental chemicals on the population in general. For reasons that are not always clear, this potential has not been realized. These exposed populations represent an underused asset in the study of the human health effects of environmental contaminants. Chronic effects thought to involve pesticides include carcinogenesis, neurotoxicity, and reproductive and development effects. In this paper we attempt to summarize this concern and, relying to a large extent on studies in our own laboratory, to indicate the importance and present status of studies of the mammalian metabolism of pesticides and indicate the need for further use of this model. Aspects considered include the role of pesticides as substrates for xenobiotic-metabolizing enzymes such as cytochrome P450 and the flavin-containing monooxygenase and their role as inducers or inhibitors of metabolic enzymes. The interaction of pesticides with complex multienzyme pathways, the role of biological characteristics, particularly gender, in pesticide metabolism, and the special role of pesticides at portals of entry and in target tissues are also considered. PMID- 8722115 TI - The road to embryologically based dose-response models. AB - The goal of researchers working in the area of developmental toxicology is to prevent adverse reproductive outcomes (early pregnancy loss, birth defects, reduced birth weight, and altered functional development) in humans due to exposures to environmental contaminants, therapeutic drugs, and other factors. To best achieve that goal, it is important that relevant information be gathered and assimilated in the risk assessment process. One of the major challenges of improved risk assessment is to better use all pertinent biological and mechanistic information. This may be done qualitatively (e.g., demonstrating that the experimental model is not appropriate for extrapolation purposes); semiquantitatively (using information to reduce the degree of uncertainty present under default extrapolation procedures), or quantitatively (formally describing the relationships between exposure and adverse outcome in mathematical forms, including components that directly reflect individual steps in the overall progression of toxicity). In this paper we review the recent advances in the risk assessment process for developmental toxicants and hypothesize on future directions that may revolutionize our thinking in this area. The road to these changes sometimes appears to be a well-mapped course on a relatively smooth surface; at other times the path is bumpy and obscure, while at still other times it is only a wish in the eye of the engineer to cross an uncharted and rugged environment. PMID- 8722116 TI - Implications for risk assessment of suggested nongenotoxic mechanisms of chemical carcinogenesis. AB - Nongenotoxic carcinogens are chemicals that induce neoplasia without it or its metabolites reacting directly with DNA. Chemicals classified as nongenotoxic carcinogens have been assumed to act as tumor promoters and exhibit threshold tumor dose-responses. This is in contrast to genotoxic carcinogens that are DNA reactive, act as tumor initiators, and are assumed to exhibit proportional responses at low doses. In this perspective, we examine the basic tenets and utility of this classification for evaluating human cancer risk. Two classes of so-called nongenotoxic chemical carcinogens selected for review include cytotoxic agents that induce regenerative hyperplasia (trihalomethanes and inducers of alpha 2-microglobulin nephropathy) and agents that act via receptor-mediated mechanisms (peroxisome proliferators and dioxin). Major conclusions of this review include: a) many chemicals considered to be nongenotoxic carcinogens actually possess certain genotoxic activities, and limiting evaluations of carcinogenicity to their nongenotoxic effects can be misleading; b) some nongenotoxic activities may cause oxidative DNA damage and thereby initiate carcinogenesis; c) although cell replication is involved in tumor development, cytotoxicity and mitogenesis do not reliably predict carcinogenesis; d) a threshold tumor response is not an inevitable result of a receptor-mediated mechanism. There are insufficient data on the chemicals reviewed here to justify treating their carcinogenic effects in animals as irrelevant for evaluating human risk. Research findings that characterize the multiple mechanisms of chemical carcinogenesis should be used quantitatively to clarify human dose-response relationships, leading to improved scientifically based public health decisions. Excessive reliance on oversimplified classification schemes that do not consider all potential contributing effects of a toxicant can obscure the actual causal relationships between exposure and cancer outcome. PMID- 8722117 TI - The melatonin hypothesis: electric power and breast cancer. AB - Breast cancer is a disease of modern life. As societies industrialize, risk increases, yet it is unclear which of the myriad changes coming with industrialization drives this increase. One important hallmark of modern life is the pervasive use of electric power. Electric power produces light at night (LAN) and electric and magnetic fields (EMF), either or both of which may alter pineal function and its primary hormone melatonin, thereby, perhaps increasing the risk of breast cancer. This hypothesis, stated a decade ago, is now receiving considerable experimental and epidemiological attention. The circumstantial case for the hypothesis has three aspects: light effects on melatonin, EMF effects on melatonin, and melatonin effects on breast cancer. The strongest of these aspects is the effects of light on melatonin. It is clear that the normal nocturnal melatonin rise in humans can be suppressed by light of sufficient intensity. The evidence for an effect of melatonin on breast cancer in experimental animals is strong, but the evidence in humans is scant and difficult to gather. The weakest aspect of the circumstantial case is EMF effects on melatonin. Whereas a half dozen independent laboratories have published findings of suppression in animals, there are inconsistencies, and there are no published data on humans. The direct evidence bearing on the hypothesis is sparse but provocative. Two laboratories have published data showing substantial increases in chemically induced breast cancer in rats by a weak AC (alternating current) magnetic field. The epidemiological evidence is very limited but has offered some support as well. An effect of electric power on breast cancer would have profound implications, and this possibility deserves continued investigation. PMID- 8722119 TI - Apparent stimulation of thrombopoesis with interleukin-3 (IL-3) in chemotherapy induced myelosuppression. AB - Two patients with advanced ovarian cancer (FIGO III C) and one patient with abdominal cancer developed severe thrombocytopenia with petechial haemorrhages after chemotherapy. These patients were treated with recombinant human interleukin-3 (rhIL-3) after multiple platelet transfusions (HLA-matched) and erythrocyte transfusions had not been effective. Increase of thrombocyte counts began three days after rhIL-3 therapy. Side effects due to rhIL-3 administration (flu-like symptoms) were moderate and could be treated effectively with paracetamol. These data support evidence for efficacy of rhIL-3 in chemotherapy induced thrombocytopenia and may open new therapeutic perspectives. PMID- 8722118 TI - Unraveling the chronic toxicity of lead: an essential priority for environmental health. AB - Although population exposure to lead has declined, chronic lead toxicity remains a major public health problem in the United States affecting millions of children and adults. Important gaps exist in knowledge of the pathophysiology of chronic lead intoxication. These gaps have impeded development of control strategies. To close current gaps in knowledge of chronic lead toxicity, we propose an integrated, multidisciplinary, marker-based research program. This program combines a) direct measurement of individual lead burden by 109Cd X-ray fluorescence analysis of lead in bone, b) determination of ALA-D phenotype, an index of individual susceptibility to lead, and c) assessments of subclinical injury produced by lead in the kidneys, nervous system and, reproductive organs. Data from this research will provide answers to questions of great public health importance: a) Are current environmental and occupational standards adequate to prevent chronic lead intoxication? b) is lead mobilized from the skeleton during pregnancy or lactation to cause fetal toxicity? c) Is lead mobilized from bone during menopause to cause neurotoxicity? d) What is the significance of genetic variation in determining susceptibility to lead? e) What is the contribution of lead to hypertension, renal disease, chronic neurodegenerative disease or declining sperm counts? f) Is chelation therapy effective in reducing body lead burden in persons with chronic overexposure to lead? PMID- 8722120 TI - Characterization of imidazopyrroloquinoline compounds synthesized from coenzyme PQQ and various amino acids. AB - Imidazopyrroloquinoline (IPQ) compounds synthesized from coenzyme PQQ (pyrroloquinoline quinone) and various kinds of amino acids were characterized. IPQ was synthesized from coenzyme PQQ and one of three amino acids (glycine, L tryptophan and L-tyrosine). Two kinds of IPQ compound were formed from coenzyme PQQ and L-serine. At higher pH, IPQ was synthesized, at lower pH hydroxy-methyl IPQ was synthesized. IPQ with R of amino acids represented by R-CH(NH2)-COOH was synthesized from coenzyme PQQ and other amino acids. IPQ compounds do not act as cofactor of glucose dehydrogenase. Molar absorption coefficients of IPQ are 31,400 M-1 cm-1 at 251 nm, 30,700 M-1 cm-1 at 276 nm, and 20,700 M-1 cm-1 at 423 nm at pH 7.0 and room temperature. PMID- 8722121 TI - A carbon-skeleton walk: a novel double rearrangement of glutaryl-CoA catalyzed by the human methylmalonyl-CoA mutase. AB - Methylmalonyl-CoA mutase is a member of the coenzyme B12-dependent family of isomerases and interconverts methylmalonyl-CoA and succinyl-CoA. We have examined the ability of the enzyme to effect a double rearrangement reaction when presented with glutaryl-CoA, a substrate analog with a three-carbon template on which two successive 1,2 migrations can occur. Our results demonstrate that the enzyme converts glutaryl-CoA to both methylsuccinyl-CoA and ethylmalonyl-CoA. To our knowledge, this is the first example of a double rearrangement reaction catalyzed by a coenzyme B12-dependent enzyme. PMID- 8722122 TI - FMN cofactor dissociation from the soluble hydrogenase of Alcaligenes eutrophus H16. AB - The specific activity of purified soluble hydrogenase of Alcaligenes eutrophus H16 was found to vary with enzyme concentration. Specific activity as a function of concentration of purified enzyme could be fit to an equation describing the dissociation of a compound into two components. An association constant, kappa(a), was determined in this way to be 39.4 +/- 8.7 micrograms protein/ml. The concentration of the enzyme affected its kinetic parameters: a tenfold decrease in enzyme concentration caused by a reduction of the V(max) and Kappa(m) (NAD) values to 45% and 58%, respectively, of the values for undiluted (0.64 mg/ml) enzyme. Diaphorase (NAD-dependent reduction of benzyl viologen) specific activity of the hydrogenase was unaffected by dilution. The extent of dilution induced activity loss was dependent on pH, with greater activity loss observed at higher pH values. The substrate NAD prevented loss of specific activity due to dilution, while the product NADH did not. Specific activity loss due to dilution as reversed with the addition of the cofactor FMN. Dilution of the hydrogenase caused an increase in the enzyme's specific flavin fluorescence. These results suggest that dilution of the soluble hydrogenase of Alcaligenes eutrophus causes dissociation of the cofactor FMN, and this activity loss should be taken into account as an important factor governing hydrogenase activity and kinetic properties. PMID- 8722123 TI - Identification of molybdopterins in molybdenum- and selenium-containing enzymes. AB - Selenium is coordinated to a molybdenum atom in nicotinic acid hydroxylase (NAH) from Clostridium barkeri and formate dehydrogenase H (FDH) from Escherichia coli. Selenium is present in FDH in a selenocysteine residue whereas in NAH in occurs in an unidentified labile cofactor. In this paper we describe a simple procedure for isolation and identification of molybdopterins from Mo-containing enzymes. The molybdopterin, after release from the protein with guanidine-hydrochloride, is reduced with KBH4, alkylated with iodoacetamide and separated on a reverse phase HPLC column. The carboxam-idomethylated pterin compound is further characterized by UV spectroscopy and mass-spectrometry. We found that FDH contains molybdopterin guanine dinucleotide whereas NAH contains molybdopterin cytosine dinucleotide. PMID- 8722124 TI - The mutual sparing effects of selenium and vitamin E in animal nutrition may be further explained by the discovery that mammalian thioredoxin reductase is a selenoenzyme. AB - The recent discovery that mammalian thioredoxin reductase is a selenoprotein furnishes an additional explanation of the mutual sparing roles of selenium and vitamin E in cellular antioxidant systems. Thioredoxin reductases isolated from human lung adenocarcinoma cells, human Jurkat T-cells and HeLa cells contain selenocysteine which is located in a C-terminal tripeptide, -Cys-SeCys-Gly. PMID- 8722125 TI - Medial compartment arthrosis of the knee. AB - When the resultant forces on the tibial plateau are displaced medially, compressive stresses cause apposition of bony tissue, thus thickening the dense subchondral bone underlying the medial plateau. Loss of the articular cartilage and an increase in subchondral bone density facilitate the progression of osteoarthrosis. Surgical management is dependent on the presence of a varus deformity; patients with medial compartment disease and varus alignment should be considered for high tibial osteotomy (HTO) or unicondylar or total knee arthroplasty (TKA), depending on their age and activity level. Patients without varus deformity and with mechanical symptoms, only mild joint-space narrowing, and pain less than 1 year are likely to benefit from arthroscopic debridement. Patients without varus alignment but with chronic pain associated with loading and more pronounced joint-space loss should be considered for HTO, or unicondylar or TKA. PMID- 8722127 TI - The frequency of and relationship between the palmaris longus and plantaris tendons. AB - A study of 186 cadavers found that 156 cadavers (83.9%) had both palmaris and both plantaris tendons. The palmaris was absent in 22 cadavers (12%); 12 cadavers (6%) were missing the plantaris tendon. If the tendon was missing from one side, there was a 67% chance that the contralateral tendon would also be missing. In all instances where both a palmaris and a plantaris tendon were absent (2.2%), the tendons were missing bilaterally. Fourteen cadavers (7.5%) missing bilateral palmaris tendons still had bilateral plantaris tendons, and 2 cadavers (1%) had no plantaris but had both palmaris tendons. Of the cadavers that had plantaris tendons, 11% had tendons that were 2 mm or less in width and of questionable value for grafting purposes. No correlation was noted between the size of the plantaris tendon and the size of the individual or the size of the palmaris tendon. PMID- 8722126 TI - 100-year-old patient with pressfit prosthesis: a postmortem retrieval study. AB - A 100-year-old woman underwent a pressfit hemiarthroplasty for treatment of a hip fracture. She was ambulant, pain free, and died 45 months after surgery. Her femurs were retrieved for analysis. Clinical, radiographic, histologic, and densitometric evaluations suggested stable bone fixation. Histologically, intimate bone contact with the prosthesis ranged from 14% to 25% over six transverse sections throughout the length of the component. Bone densitometry was generally equal to the nonimplanted femur, except at the medial cortex. Few osteoclasts or inflammatory cells were noted. Most new bone was noted at the corners of the implant. These results suggest that cementless pressfit fixation may be achievable in elderly patients and that osteointegration and prosthetic stability is possible without a complete "fit and fill." PMID- 8722128 TI - A rapid method for emergency ring removal. AB - Injuries caused by embedded rings are extremely rare. Swelling can lead to decreased circulation and can threaten the viability of involved fingers. Skin changes can become chronic, as when rings are left on fingers of patients with decreased mentation or sensation. In this report, the authors discuss a technique for removal of embedded rings from the fingers of a schizophrenic patient. PMID- 8722129 TI - Ganglion of the anterior cruciate ligament presented as a knee mass. AB - A ganglion cyst of the anterior cruciate ligament (ACL) is a rare disorder and should be considered in cases of benign masses of the knee. A ganglion cyst originating on the ACL found at surgery on a 26-year-old woman was not diagnosed during her preoperative evaluation. PMID- 8722130 TI - An unusual fracture dislocation pattern in a patient with an Os lunatotriquetrum. PMID- 8722131 TI - Treatment of high-energy proximal tibial fractures using the Monticelli-Spinelli external fixator: a preliminary report. AB - Between 1990 to 1993, 21 patients with tibial plateau or proximal tibial fractures resulting from high-energy trauma were treated with the Monticelli Spinelli external fixator. There were 13 men and 8 women (mean age, 45.2 years; range, 26 to 78). There were a total of 5 type A, 2 type B, and 14 type C fractures, using the Arbeitsgemeinschaft Fur Osteosynthesefragen (AO) classification system. Immediate postoperative reductions were good or excellent, according to strict radiographic criteria, in 16 of 21 patients. All patients obtained at least 90 degrees of knee flexion, and only one patient lost more than 5 degrees of full extension. Complications included seven superficial pin-tract infections and one deep vein thrombosis with resultant pulmonary embolism. Nineteen patients were available for follow-up (mean, 14 months). Clinically, 13 patients had satisfactory results where good or excellent radiographic reductions were maintained, knee extension was within 5 degrees of full, flexion was > or = 90 degrees, with < or = 5 degrees valgus/varus angulation. Six patients had unsatisfactory results, not meeting the above criteria. The Monticelli-Spinelli external fixator is a much-needed tool in the treatment of high-energy tibial plateau fractures that are not amenable to more extensive surgical procedures because of the associated soft-tissue injuries. PMID- 8722132 TI - A 68-year-old man with gradual onset of low-back pain. AB - The following case is presented to illustrate the roentgenographic and clinical findings of a condition of interest to orthopedic surgeons. The initial history, physical findings, and roentgenographic examinations are noted on this page. The clinical and roentgenographic diagnoses are presented on the following pages. PMID- 8722133 TI - A technique for comparison of leg lengths during total hip replacement. AB - Leg-length inequality after total hip arthroplasty is not uncommon and can be distressing to the patient. The use of a particular design of Steinmann pin for comparing a patient's leg lengths during total hip arthroplasty is illustrated. PMID- 8722134 TI - Primary health care and severe mental illness: the need for national and state policy. PMID- 8722135 TI - Primary health care physicians' treatment of psychosocial problems: implications for social work. AB - This study explores the extent to which primary care physicians serve as gatekeepers for the treatment of psychosocial problems and the extent to which they have contact with social workers. We also attempted to identify physician variables related to gatekeeping and amount of contact with social workers. Data from a national study of the role of primary care physicians in Israel were used. About one-third of the physicians had regular contact with social workers, one quarter had contact less often, and about one-third almost never had contact. Variables associated with gatekeeping for psychosocial problems included having a large caseload, specializing in family medicine, practicing in a rural location, and being under age 55. Variables associated with social work contact were affiliation with the largest health maintenance organization and specialization in family medicine. PMID- 8722136 TI - Discharge planning for dementia patients: factors influencing caregiver decisions and satisfaction. AB - Discharge planning is a major concern of hospitals as they seek to reduce costs associated with prolonged lengths of hospital stays, prevent subsequent readmissions, and ensure that discharge dispositions are appropriate. This process can be even more complicated when the patient is suffering from dementia and the family has primary responsibility for the discharge outcome. Data on the discharge process and caregiver satisfaction with it were collected from 179 caregivers of hospitalized dementia patients. The findings indicate that social workers are important influences in the discharge decisions but are significantly more influential when the discharge is to a nursing home. Findings from regression analyses indicate that caregiver satisfaction is dependent on the amount of caregiver involvement in the process. PMID- 8722138 TI - Social work and end-of-life decisions: self-determination and the common good. AB - Client self-determination is the key element of NASW's policy statement about social work intervention in end-of-life decisions. However, both self determination and the common good must be respected in social work practice and policy regarding end-of-life decisions. This article discusses self-determination in end-of-life decision making, ethical decision making and the NASW Code of Ethics, and professional ethics based on a balanced view of both self determination and the common good. Recommendations for professional practice and social policy are offered. PMID- 8722137 TI - Immunization among African American children: implications for social work. AB - Although childhood immunizations have proved to be one of the most effective means of preventing and controlling the spread of infectious and communicable diseases, thousands of preschool children, particularly children from urban African American poor families, are not being immunized. This article discusses the critical problem of low rates of immunization for this population and identifies ways social workers can play an active role in ensuring that more children are immunized. Immunization of preschool children is a function of the interrelationship among health-seeking behavior of parents, financial and nonfinancial barriers to health care, and provider practices that inhibit appropriate immunization. Improving access to existing public programs, facilitating community organization efforts, assisting communities through self help and mutual-aid initiatives, and supporting national efforts can improve immunization status among poor children. PMID- 8722139 TI - Attitudes toward and knowledge of chronic pain: a survey of medical social workers. AB - This study addressed two research questions: What are medical social workers' attitudes toward the management of chronic pain, and how knowledgeable are they about chronic pain? Findings indicate that social workers hold a highly positive orientation toward both nonmedical treatment of chronic pain and social work involvement in chronic pain services, but they display low levels of knowledge about chronic pain and pain management. Together these findings suggest that social workers possess the willingness and opportunity to develop a social work specialization in pain management but may be hindered by an uncertain knowledge base in the area. PMID- 8722140 TI - Racial differences in discharge planning. AB - Given previously reported findings of racial differences in elderly people's use of posthospital care, this article focuses on discharge planning processes as explanations of differential service utilization. We studied the discharge plans for 369 African American and white elderly patients and examined options pursued for posthospital care by social workers, patients, and families for evidence of racial differences. We also looked for racial differences in ruling out nursing home care for reasons of patient and family preference. Discharge planning with African American patients and family members involved less pursuit of nursing home care and more pursuit of formal services in the home than planning with white patients and families. Implications for practice and future research are discussed. PMID- 8722141 TI - Gay men with AIDS and their families of origin: an analysis of social support. AB - This article reviews the literature on the relationships between gay men with AIDS and their families of origin to determine why the family is not a principal source of social support. Several reasons explain the absence of the family from the support network, including the family's lack of acceptance of homosexuality and the relationship with a male partner; the stigma associated with AIDS; the inability of family members to communicate openly about homosexuality and AIDS; the lack of competence among family members in dealing with HIV issues; and overprotective, infantilizing behavior by parents. A sibling, most often a sister, is the family member to whom the gay man with AIDS feels closest and from whom he is most likely to seek support. Implications for practice and research are discussed. PMID- 8722142 TI - The governors' dangerous Medicaid endgame. PMID- 8722143 TI - [The surgical treatment of pancreatic cysts and pseudocysts]. AB - Pancreatic cysts and pseudocysts are now being detected more often as a result of wide use of abdominal echography and CT scan. Surgical strategy advocates resection whenever possible for true cysts and internal drainage for pseudocysts. The present study retrospectively analyses 8 cases of true cysts and 23 cases of pseudocysts submitted to surgery in a twenty years period. Conservative treatment and ultrasonography-guided percutaneous drainage were not discussed. Four true cysts with positive benign structure were treated by internal drainage and only two adenocarcinomas were resected. In two more malignancies internal drainage was done as a palliative procedure followed by lethal outcome. Pseudocysts were secondary to acute pancreatitis in 7 cases, chronic pancreatitis in 6 cases, abdominal trauma in 4 cases and were idiopathic in 6 cases. Caudal pancreatectomy was performed whenever distal localisation was diagnosed. External drainage imposed in 5 emergency situations but internal drainage was elective in 13 cases. In conclusion according to our experience internal drainage should be preferred as the main surgical treatment in all mature pseudocysts and also when benign true cysts are certified. Posterior transgastric cystogastroanastomosis is the procedure of choice. PMID- 8722144 TI - [A comparative study of the hospital costs for laparoscopic cholecystectomy and classical cholecystectomy]. PMID- 8722145 TI - [Training in laparoscopic surgery. The problem of competence and responsibility]. AB - Laparoscopic cholecystectomy has become the procedure of choice for the treatment of gallbladder disease. Many general surgeons have incorporated laparoscopic cholecystectomy into their clinical practices usually after completing postgraduate didactic and laboratory animal training course. As in laparoscopic surgery, it is necessary a special training of the surgeons, it is recommended that the medical universities for postgraduate studies should train them in this respect. At the same time the medical scientific societies should confirm the competence the surgeons' competence in the field and, moreover protect them legally. PMID- 8722146 TI - [Diagnostic and therapeutic endoscopic retrograde cholangiopancreatography, an analysis of 100 consecutive cases]. PMID- 8722147 TI - [Post-bulbar duodenal ulcer]. PMID- 8722148 TI - [Intra-abdominal fibromatosis]. AB - The authors describe three cases of intraabdominal fibromathosis: two cases with intraperithoneal location and another one with retroperithoneal location. All of them are benign noncapsulated tumours of the fibrous tissue with tendancy to local recurrence. Abdominal fibromathosis may determine any form of acute or chronic digestive manifestations. Only to the accuracy of the histo-pathological examination is due the diagnosis between fibromathosis and fibrosarcoma, reactive fibrosis, mixoma and nodular fasceitis. The surgical excision must not be economical and the association with radiant therapy must also be considered. PMID- 8722149 TI - [Is medical/surgical false acute abdomen a clinical reality?]. AB - The false acute abdomen is a real Clinic picture. In pur experience it represents the 0.63% of acute surgical abdomen. Sometimes diagnosis could escage because patient's erethism. To obviate this event surgeons must reach a detailed history, perform a complete pshysical examination and repeat clinical evaluation many times above all when then the clinical picture is dubious. Laboratory and instrumental tests could be useful to reach accurate diagnosis. However, in our opinion, when clinical and physical examination are unclear, sometimes in spite of disappointment of other physicians, do explorative laparotomy is better is than non-treatment of an acute surgical abdomen. False acute abdomen. PMID- 8722150 TI - [Fibroma of the ascending colon]. PMID- 8722151 TI - [Familial abdominal fibromatosis]. AB - Two cases of abdominal fibromatosis are followed-up in two brothers patients development, each of them having a peculiar development. First of them underwent operation for a huge abdominal tumor with a retroperitoneal origin and intraperitoneal development which needed a complex partial resection with first jejunal loop enterectomy. The other patient had first surgery for fibrosarcoma of nuchal area and after that he underwent an operation for superior digestive haemorrhagia as a result of antral gastric fibroid tumor with transverse colic and mezocolic extension, which needed gastro-colectomy. The patients father was followed up for tangible abdominal tumors, but he rejected the proposed coeliotomia. The two brothers patients had a good postoperative development. The examination of the charriotype showed anomalies of the short branch of the 21st and 22nd chromosome (which are still normal). PMID- 8722152 TI - [Pancreatic hydatid cyst]. PMID- 8722153 TI - [AIDS and tuberculosis]. PMID- 8722154 TI - [Use of antibiotics in primary care: treatment of urinary infection]. AB - OBJECTIVES: To analyse the primary care therapeutic approach to urinary tract infections (UTI), and, a secondary objective, to obtain information on the microbiological profile and resistance to antibiotics in this field, with the aim of making therapeutic recommendations. DESIGN: The UTI diagnosed through systematic checking of sediment and/or culture, requests from health centres are described. SETTING: 9 Health Centres in Palma (Mallorca) during November 1992. PATIENTS AND OTHER PARTICIPANTS: Out of 2,484 requests for sediment and/or uroculture, clinical history was found in 2,033. At the end there were 232 patients whose request for analysis was for suspected UTI, among whom there were 43 recurring cases, which meant a total of 275 cases. MEASUREMENTS AND MAIN RESULTS: The most frequent germ was Escherichia coli. 68% had complicated UTI. Empirical treatment was carried out in 60.7%. Treatment of the first episode of UTI was with quinolones in 73.3% of cases, with norfloxacine in 44.1%. There was a higher percentage of resistance of Escherichia coli to cotrimoxazole (49.0%), ampicilline-sulbactam (46.4%), ampicilline (42.0%) and cephalexin (31.4%). In 50 cases there was therapeutic failure, due (among the most commonly used drugs) to norfloxacine in 12.4%, to cotrimoxazole in 28.6%, to norfloxacine in 27.3% and to pipemidic in 18.9%. CONCLUSIONS: There are many more specific than empirical treatments as well as excessive use of drugs not recommended as first choice in primary care. The level of resistance to the new quinolones is considerable and is greater still for some antimicrobial drugs used for UTI. PMID- 8722155 TI - [Improvement in the quality of the interclinic referral note in regard to the information given]. AB - OBJECTIVE: To quantify the improvement in compliance with filling out the interclinical note after both general practitioners and specialists were informed of the compliance level found in an earlier study. DESIGN: An intervention before and-after study. Unit of analysis: interclinical note in the primary care clinical record. SETTING: Health Areas 11 and 12 in the Community of Valencia, November 1993 and 1994. Field work was developed on the premises of the two specialist centres of these areas. PARTICIPANTS: Two samples, of 708 and 326 interclinical notes, were used. These were stratified in function of the number of consultations during the first six months of each of the study years, by centre and speciality. INTERVENTION: All primary care and specialist professionals were informed of the level of compliance with the interclinical note in the first part of the study. MAIN RESULTS: The primary care section improved significantly in: personal history, data on physical examination, suspected diagnosis, reason for consultation. The section on specialists improved significantly in the identification of the specialist, diagnosis, treatment, action to take. CONCLUSIONS: Information given to professionals improved their filling out of the interclinical note, basically on variables to do with the pathological process. It should be routine to feed back information to the professionals concerned. PMID- 8722156 TI - [Analysis of information sources in the journal, Atencion Primaria]. AB - OBJECTIVES: To contribute to improving the understanding of the information bases of Spanish scientific production in the primary care field; to aid international contacts and the diffusion of information among doctors in our country. DESIGN: A bibliometric analysis of the bibliographic references of the studies published in the journal Atencion Primaria during 1991. A data base, managed by dBASE IV, was created. SETTING: The Unit of Documental Analysis and Bibliometry of the Institute of Documental and Historical Studies in Science (University of Valencia CSIC). MATERIAL: 2,615 bibliographic references in 205 studies contained in volume 8 of Atencion Primaria. MAIN RESULTS: References from journal articles (68%) and books (26%) predominated. There were a large number of references from Spanish publications (45%) as against Northamerican (27%) and British (15%). There were few references from other European countries or Latin America. Information was quickly out of date (4 year semiperiod; Price index 50%). High proportion of self-quotations from the journal. CONCLUSIONS: The major role of the journal Atencion Primaria in communicating information in the primary care field was underlined. The small amount of information from Latin America and E.U. countries (except Great Britain) highlights the isolation of Spain from these countries, which can be explained by the limits of the primary care field itself. There was also little use of information to which access was more difficult for primary care doctors, such as doctoral theses, congress papers or reports. Moreover this is a field with a high percentage of recent literature and rapidly out-of-date information. PMID- 8722157 TI - [Perception of the risk of venous thromboembolic disease in primary care]. AB - OBJECTIVES: To determine the prevalence in out-patients of risk factors leading to venous thromboembolic disease (VTE); and to analyse what perception primary care centre (PCC) doctors have of this risk and what attitude they adopt to therapy. DESIGN: 1) An observational, crossover study, with patients included at random. 2) A study with a teaching intervention, which was neither controlled nor randomised. PATIENTS: PCC patients over 25 seen either on demand, with appointments or at home. INTERVENTION: A clinical history was composed, using a closed questionnaire. A risk category and therapeutic attitude were then assigned. A guide on prevention and treatment of VTD was presented and discussed in clinical sessions. RESULTS: 11 PCC doctors polled 272 patients. 45 of these had moderate to high risk of VTD; and 4 had one of the criteria of the European Accord on VTD prevention for commencing prophylactic treatment. In two of these four, it was thought necessary to start prophylaxis. CONCLUSIONS: The prevalence of patients with moderate to high risk of VTD is 17% in patients over 25 who seek medical attention at PCCs. Doctors detected 50% of moderate to high risk patients. If the criteria used for in-patients were followed, prophylaxis should have been started in 1.5% of patients. PMID- 8722158 TI - [Factors affecting the follow up of hypertensive patients]. AB - OBJECTIVES: To study the factors affecting lack of continuity in hypertense patients' follow-up care and to define these patients' profile. DESIGN: Crossover study. SETTING: Rural health district. PATIENTS: Systematic sample of the 1,487 hypertense patients in our health district (n = 642). INTERVENTIONS: Patients' socio-demographic variables, characteristics of their AHT, treatment, risk factors and chronic illnesses were taken from the clinical records. The following were noted concerning lapses (defined as periods longer than six months without a blood pressure reading): beginning, end and number of lapses, consultations during a lapse and reasons for the consultation. RESULTS: 56% of the individuals in the sample had lapses. These patients consulted 2.8 times during each lapse, making a total of 1,174 consultations (47.1% acute problems; 23.5% chronic pathologies; 19.8% administrative matters and 7.5% AHT or related ailments). On comparing patients with and without lapses we found that variables with significant differences were: pensioner, education, profession, initial and current pharmacological treatment, smoker, years of the AHT's evolution and doctor. No significant differences were found for age, gender or tension figures. Lapses were less in diabetics and patients with Hypercholesterolaemia. CONCLUSIONS: Although we defined an approximate profile of the patient with lapses, we found no relationship with tension figures. Both the fact that a high number of patients consult during lapses and the differences between clinics show the co-responsibility of the health factor as a cause of lack of continuity. PMID- 8722159 TI - [Apgar questionnaire in the study of family function]. AB - OBJECTIVES: To analyse family dynamics among health service users. To establish a connection between perception of family dysfunction and different variables. DESIGN: A descriptive crossover study, in line with a closed question survey model: a) family Apgar, b) 16 item questionnaire. SETTING: Jumilla Health Centre, Murcia. PATIENTS: 397 patients between 18 and 65 were selected by systematic sampling. INTERVENTIONS: The Chi-squared statistical method and remainder analysis were used. MEASUREMENTS AND RESULTS: a) Analysis of family perception: 315 normally functioning (79.35%) and 82 dysfunctional (20.65%). b) There were statistically significant connections between family dysfunction and the number of brothers and sisters and the place occupied by them, marital status, number of children, educational level, mental health and reason for the consultation. CONCLUSIONS: The family Apgar enables patients with family dysfunction to be identified. There is a clear connection between family dysfunction and the reasons for seeking health care. PMID- 8722160 TI - [Diagnosis of mild arterial hypertension: 3 or 5 measurements?]. AB - OBJECTIVE: To determine the differences in the diagnosis of light arterial hypertension between using three tension measurements (over 2 or 3 weeks) or five (6 or 7 weeks). DESIGN: A descriptive, longitudinal study. SETTING: Santomera Health Centre, Murcia. PARTICIPANTS: In 245 patients initially diagnosed as suffering from light Hypertension (with three takes of blood pressure), two more tension measurements were taken, over four weeks, to make a definitive diagnosis of light arterial Hypertension. 156 patients completed the series, for whom the differences in diagnosis between three and five takes, and variations in their blood pressure figures on repetition of the measurements, were studied. There were no differences in patients' basic characteristics between those who completed the tests and those who didn't. But there were differences between different professionals regarding their compliance with the procedure. RESULTS: 19.9% of the patients diagnosed initially after three blood pressure measurements did not have the diagnosis confirmed after five. Repetition of the measurements signified decrease in diastolic (p < 0.00005) and systolic (p < 0.00005) pressures. Average age of patients definitely diagnosed as having light hypertension was higher than the age of those not confirmed (p < 0.005). The older the patient, the higher the systolic pressure (p < 0.0001). CONCLUSIONS: The diagnosis of light arterial hypertension differed in 19.9% between three measurements and five. PMID- 8722161 TI - -Factors of dissatisfaction with health care. A Delphi study-. AB - OBJECTIVE: To study the causes of the disagreement between the general atmosphere of insatisfaction inherent in the assistance offered by the health care system and the favourable evaluation that the public shows in the questionnaires carried out in the field of primary health care. The less satisfactory issues of the health service offert are also identified. DESIGN: A Delphi thecnique has been used as consensus method. SETTING: Community setting: Salamanca urban area. PARTICIPANTS: The Delphi group is made up of people working in jobs with a pronounced social character: members of neighbour associations, educators, social workers, etc. Three consecutive questionnaires were distributed among fifty people. Twenty-seven of them completed the three rounds. MEASUREMENTS AND MAIN RESULTS: The score of the reasons of disagreement perceived as the most important were: the difficulties of access at a specialised level, 8.3 points; the inequality of the services offered by the two organizative models that coexist in the primary health care, 7.1; the lack of discrimination between the primary and the specialized level, 6.8. The highest scores were for the following complaints: excessive waiting lists at the second care level (8.5), delay in solving the problems (7.8) and masification at the consultancy (7.1). The most satisfactory aspects are the possibility of free choice of doctor, 7.9 points; the previous appointement system, 7.6; and a reliable relationship with the doctor, 7.3. CONCLUSIONS: To take into account these factors in the future design and interpretation of inquiries will increase their information validity and usefulness. PMID- 8722162 TI - [Examinations and referrals at a health center: a study of associated factors]. AB - OBJECTIVES: To determine the profile of the use of physical examination, further tests and referrals; and the impact which socio-family and behavioural factors, questions connected with patients' morbidity and other variables in health service use, have on these resources. DESIGN: Descriptive crossover study. SETTING: Urban health centre. PATIENTS AND OTHER PARTICIPANTS: 719 attendances of patients seen over a two and a half month period in two general medical clinics at the health centre were analysed. A questionnaire on health care was presented by the doctor to the patient at the end of the visit and also covered the activities and resources used during the visit. MAIN RESULTS: A multivariant analysis identified the factors which best explained variability in the frequency of physical examination as symptoms and, in particular, the type of consultation. None of the factors studied affected the use of analysis, radiology or referral to specialists. CONCLUSIONS: Symptoms and, in particular, the type of consultation are the factors which affect the use of physical examination. The scant relevance of other factors analysed and the comparison of our findings with other studies published suggest that the use of further examinations and referrals depend basically on other variables, such as those connected to the availability of resources and accessibility to them. PMID- 8722164 TI - [Use of ticlopidine in primary care]. PMID- 8722163 TI - [Human scabies]. PMID- 8722165 TI - [Brucellosis in a low incidence zone]. PMID- 8722166 TI - [Myasthenia gravis caused by D-penicillamine]. PMID- 8722167 TI - [Chronic lymphatic B-cell leukemia. Clinical approach in primary care]. PMID- 8722168 TI - Cutting apart V(D)J recombination. AB - The past year has seen major advances in our understanding of the recombination mechanism by which antibody and T cell receptor genes are assembled during lymphoid development. The initial cleavage events can be carried out in vitro by purified RAG1 and RAG/ protein. In addition, a number of genes involved in later steps of the reaction have been cloned, opening the way for an in-depth biochemical analysis of this critical developmental process. PMID- 8722169 TI - DNA repair and transcription. AB - The transcription factor TFIIH continues to be a subject of interest. In addition to its function as a repair and transcription factor, TFIIH includes a cyclin dependent kinase and a cyclin, which raises the possibility that nucleotide excision repair (NER), RNA polymerase II transcription and cell cycle control are connected. Progress in mechanistic studies of NER include the identification of dual incision activities operating on either side of base damage and the isolation of a repairosome supercomplex in yeast. Additionally, NER has been demonstrated in reconstituted human and yeast systems, both of which include TFIIH. PMID- 8722170 TI - Transcription: basal factors and activation. AB - Much progress has been made in the past few years in understanding the mechanism and regulation of mRNA synthesis. This rapid progress has largely been due to the availability of cloned genes encoding components of the transcription machinery. Structural and biochemical studies are rapidly defining the architecture of components in the transcription complex. Highly purified biochemical systems are beginning to elucidate the role of the individual initiation factors. The identification of a large complex that contains a polymerase, termed holoenzyme, has provided a new way of thinking about how the transcription complex assembles at a promoter. The mechanism of transcription stimulation by activators is beginning to be unraveled but still appears to be a complex process. Finally, analyses of genes involved in DNA repair, cell cycle control and transcription have revealed similarities between transcription and other forms of cell regulation. PMID- 8722171 TI - Multiple roles for DNA methylation in gametic imprinting. AB - Allele-specific DNA methylation has been observed for all tested imprinted genes and has a clear role in the imprinting mechanism. It remains to be resolved whether this role is to act as the gametic imprinting signal or to cause or maintain allele-specific expression. PMID- 8722172 TI - Regulation of gene expression by nucleosomes. AB - During the past year, the characterization of mechanisms and factors capable of disrupting nucleosomes during transcriptional activation has been a recurrent theme in studies which address the contribution of nucleosome structure to gene regulation. In vivo studies using yeast and Drosophila together with biochemical purification schemes using nucleosome perturbation assays have provided evidence for the existence of multiprotein complexes that are able to alleviate nucleosome repression. At the same time, new insights into the mechanism of heterochromatin formation have been gained, which have direct links to nucleosome structure. PMID- 8722173 TI - Multiple SWItches to turn on chromatin? AB - The SWI/SNF complex is a highly conserved multisubunit assembly that facilitates the function of gene-specific transcriptional regulatory proteins by antagonizing chromatin-mediated transcriptional repression. Recent studies have suggested the existence of multiple functionally distinct SWI/SNF-like complexes. One possibility is that different chromatin remodeling systems are targeted to different gene sets or, alternatively, that they may remodel chromatin structure to facilitate cellular processes other than transcription, such as recombination or DNA repair. PMID- 8722174 TI - Special HATs for special occasions: linking histone acetylation to chromatin assembly and gene activation. AB - Post-translational acetylation of the core histone amino-terminal tails has long been associated with both chromatin assembly and the regulation of gene expression. The recent identification and cloning of histone acetyltransferase genes represents a significant breakthrough in our understanding of how specific acetylation states are established. Ongoing characterization of these enzymes and their molecular cohorts supports a direct role for acetylation in a signaling pathway that modulates chromatin structure to create new patterns of transcription. PMID- 8722175 TI - Boundary and insulator elements in chromosomes. AB - Recent progress in understanding boundary and insulator elements has concentrated on the identification of their protein components. BEAF-32 is a protein present in the scs' element of Drosophila that is also localized to most interband regions and puffs of polytene chromosomes, suggesting a role in the organization of structural chromosomal domains. The suppressor of Hairy-wing and modifier of mdg4 proteins have been characterized as components of the gypsy insulator. The latter seems to play a crucial role in conferring on the insulator its ability to unidirectionally affect enhancer function. PMID- 8722176 TI - Heterochromatin and gene regulation in Drosophila. AB - We have recently learned more about the biochemistry of heterochromatin and about how heterochromatic environments affect gene function. New findings have emphasized the distinctions between telomeric and pericentric heterochromatin in Drosophila and have suggested a mosaic structure within pericentric heterochromatin. Theories concerning the mechanism of inactivation of euchromatic genes in heterochromatic environments have been tested using transgenes inserted into heterochromatin. The current data support a competition/chromatin structure model, in which multiprotein repressor complexes compete with transcriptional activators to assemble an active or inactive chromatin structure. PMID- 8722177 TI - Replication origins in eukaroytes. AB - Recent experiments in budding yeast and Xenopus have provided new insights into the regulation of eukaroytic DNA replication. The multi-subunit origin recognition complex plays a key role in initiation, remaining bound at origins of replication during most of the cell cycle. Early in the cell cycle, Cdc6 and the Mcm proteins 'reset' chromatin for another round of DNA replication. Cyclin dependent kinases appear to play a dual role, both in activating replication origins and blocking the formation of new pre-replicative complexes; thus limiting replication to once per cell cycle. PMID- 8722178 TI - Cell cycle control of eukaryotic DNA replication. AB - A clearer picture of replication control is emerging through the characterization of proteins, such as cdc18/Cdc6 and members of the mini-chromosome maintenance (MCM) protein family, that are involved in the initiation step. Cyclin B dependent kinases have conserved roles in both Saccharomyces cerevisiae and Schizosaccharomyces pombe, switching on DNA replication in G1 and preventing re replication in G2. A model is suggested where MCMs and CDKs play complementary roles to ensure 'once-per-cell-cycle' replication, with CDKs maintaining a G1 or G2 state, whereas MCMs provide a cis-acting control on chromatin to prevent reinitiation during a single S phase. PMID- 8722179 TI - Initial splice-site recognition and pairing during pre-mRNA splicing. AB - Advances over the past year have provided new insights into the mechanisms involved in the initial recognition and pairing of the 5' and 3' splice sites in complex metazoan pre-mRNAs. Highlights include the demonstration that exonic enhancers can promote trans splicing and that an excess of the serine and arginine rich family of splicing proteins can obviate the requirement for U1 small nuclear ribonucleoprotein particle in splicing. PMID- 8722180 TI - RNA editing: how a message is changed. AB - Considerable progress has been made in unraveling the mechanistic features of RNA editing processes in a number of genetic systems. Recent highlights include the identification of the catalytic subunit of the mammalian apolipoprotein B mRNA editing enzyme as a zinc-dependent cytidine deaminase that binds to RNA, the demonstration that adenosines in brain glutamate receptor pre-mRNAs are converted into inosines and that double-stranded RNA A deaminase (dsRAD), the candidate enzyme, is another zinc-dependent RNA nucleotide deaminase, and a mounting body of evidence for a cleavage-ligation mechanism for U insertion/deletion editing in kinetoplastid protozoa. PMID- 8722181 TI - Chromosomes and expression mechanisms. PMID- 8722182 TI - Influence of pH on heat resistance of Bacillus licheniformis in buffer and homogenised foods. AB - The influence of pH of heating menstruum (McIlvaine buffer) on the heat resistance of Bacillus licheniformis was investigated and compared with the heat resistance in homogenised tomato and asparagus at pH 7 and 4 in a wide range of temperatures. Heat resistance was in all mestrua smaller at acid pH. At 99 degrees C and pH 4, heat resistance was 1/20 lower than at pH 7. However, the magnitude of this effect decreased as heat treatment temperatures were increased almost disappearing at 120 degrees C. z values increased from 6.85 at pH 7, to 10.75 at pH 4. At 99 degrees C the effect of pH on heat resistance was constant along the range of pH's tested. The increase of one pH unit increased D99 by 180%. At pH 7 and 4, heat resistance was the same in buffer as in tomato and asparagus homogenates at all temperatures tested. The diminishing influence of the acidification of some foods on the heat resistance of B. licheniformis sterilisation temperatures should be taken into account when a raise in temperature is considered to shorten the duration of heat processes. PMID- 8722183 TI - Differentiation of bifidobacteria by use of pulsed-field gel electrophoresis and polymerase chain reaction. AB - Several different genomic fingerprints can be obtained from various commercially important species of Bifidobacterium using pulsed-field gel electrophoresis (PFGE) following digestion of DNA with XbaI and SpeI. Four different genomic finger printings were discernible for reference strains of Bifidobacterium animalis, five for B. bifidum, three for B. breve, five for B. infantis and three for B. longum. Standard commercially-available industrial strains of B. animalis are identical to the reference strain ATCC 27536, previously isolated from chicken feces. There was more genomic heterogeneity among industrial strains of B. longum, in that only one gave profiles similar to the type strain of this species (ATCC 15707). The other 14 commercially-available strains of B. longum (mainly isolated from Japanese commercial preparations) were divided into four new molecular types based on their PFGE patterns. The PFGE method indicated that only five distinct strains of B. longum and one strain of B. animalis are used in commercial preparations. Additionally, the use of polymerase chain reaction amplification of portions of 16S rDNA provides a highly specific technique to discriminate between the species B. breve, B. infantis and B. longum. PMID- 8722184 TI - Salmonella detection by the polymyxin-cloth enzyme immunoassay using polyclonal and monoclonal detector antibodies. AB - Several commercially available O-antigen polyclonal antisera and a monoclonal antibody to the core region of lipopolysaccharide (LPS) were examined as sources of detector antibodies in a polymyxin-cloth enzyme immunoassay (polymyxin-CEIA) for Salmonella. In this assay, polymyxin-coated polyester cloth captured the LPS antigens from Salmonella broth cultures, followed by immunoenzymatic detection of the captured LPS using specific antibodies. Pools of polyvalent antisera reacted with all of the Salmonella strains tested, but also gave cross-reactions with some non-Salmonella bacteria. On the other hand, the monoclonal antibody gave positive reactions with all of the Salmonella tested except serogroup O-strains, but did not react with any of the non-Salmonella bacteria. The monoclonal antibody supplemented with a single factor serogroup O:35 rabbit antiserum was able to detect the serogroup O-strains without causing any cross-reactions with the non-Salmonella bacteria. As an example of the applicability of this assay system, low levels of Salmonella cells spiked into various food samples were successfully detected after an overnight enrichment in broth. PMID- 8722185 TI - Collaborative ring-trial of Dynabeads anti-Salmonella for immunomagnetic separation of stressed Salmonella cells from herbs and spices. AB - Eight laboratories participated in a Salmonella detection ring-trial which compared selective enrichment by conventional broths with immunomagnetic separation (IMS) using Dynabeads Anti-Salmonella. Laboratories analyzed six types of herbs and spices that were spiked with one of six freeze-dried Salmonella species. Each herb and spice analysis comprised of 12 samples (25 g each) which had been spiked at three different levels, plus a negative control and stored for one week prior to testing. Out of a total 468 samples analyzed, 195 (41.7%) were positive by both methods. Eighteen samples were positive only by IMS enrichment, in comparison with 19 positive samples by conventional enrichment broths and not IMS. These results confirm the potential use of IMS as an alternative to enrichment broths for Salmonella isolation. PMID- 8722186 TI - Microbiology of mesu, a traditional fermented bamboo shoot product. AB - The use of mesu as a pickle and as the base of curry is a tradition in the Darjeeling hills and Sikkim of India. A total of 327 strains of lactic acid bacteria, representing Lactobacillus plantarum, L. brevis and Pediococcus pentosaceus were isolated from 30 samples of mesu. These species were present in all samples of raw bamboo shoots tested. Mesu was dominated by L. plantarum followed by L. brevis; P. pentosaceus was isolated less frequently and recovered from only 40-50% of the mesu samples. The fermentation was initiated by P. pentosaceus, followed by L. brevis, and finally succeeded by L. plantarum species. During fermentation, the titratable acidity increased from 0.04 to 0.95%, resulting in the decline in pH from 6.4 to 3.8. PMID- 8722187 TI - Thermotolerance of meat spoilage lactic acid bacteria and their inactivation in vacuum-packaged vienna sausages. AB - Heat resistance of three meat spoilage lactic acid bacteria was determined in vitro. D-values at 57, 60 and 63 degrees C were 52.9, 39.3 and 32.5 s for Lactobacillus sake, 34.9, 31.3 and 20.2 s for Leuconostoc mesenteroides and 22.5, 15.6 and 14.4 s for Lactobacillus curvatus, respectively. The three lactic acid bacteria were heat sensitive, as one log reductions in numbers were achieved at 57 degrees C in less than 60 s. Z-values could not be accurately determined as D values did not change by a factor of 10 over the temperature range studied. In package pasteurization processes were calculated using the highest in vitro D value and applied to vacuum-packaged vienna sausages. Microbiological shelf life (time for lactic acid bacteria count to reach 5 x 10(6) CFU/g) increased from 7 days for non-pasteurized samples to 67, 99 and 119 days for samples of the three pasteurization treatments at 8 degrees C storage. Enterobacteriaceae were detected at levels of log 4.0 CFU/g in non-pasteurized samples, but were reduced to < log 1.0 CFU/g in pasteurized samples. The incidence of listeriae in non pasteurized samples was low as only one Listeria innocua strain was isolated. No Listeria spp. were isolated from pasteurized samples. Numbers of Clostridium isolates increased from one in non-pasteurized samples to 25 in pasteurized samples. Increasing incidences of clostridia, and the presence of C. perfringens in pasteurized samples indicated that in-package pasteurization could compromise product safety. PMID- 8722188 TI - Isolation of thermotolerant species of Campylobacter from commercial chicken livers. AB - The qualitative and quantitative contamination levels of Campylobacter spp. in frozen commercial chicken livers were assessed. From the 126 samples studied, 117 (92.9%) were positive for Campylobacter spp. C. coli was isolated more frequently (78.6%) than C. jejuni (21.4%). The MPN method showed that Campylobacter contamination levels ranged from 10 to > 230 bacteria/100 ml of liquid exuded by livers. PMID- 8722189 TI - Ferrioxamine E-supplemented pre-enrichment and enrichment media improve various isolation methods for Salmonella. AB - Supplementation of pre-enrichment broth and enrichment broth media with ferrioxamine E (1 microgram/ml) significantly improved the recovery of Salmonella from artificially or naturally contaminated foods. Based on the selectivity of ferrioxamine E, Salmonella enteritidis and S. typhimurium could be isolated also from various mixed cultures (one Salmonella cell in 10(3)-10(4)-fold concentration of cells of competitors) by shaking for 6 h in supplemented buffered peptone water followed by cultivation on XLD- or XLT-4 agars. Isolation of Salmonella from these pre-enrichment cultures by use of Dynabeads-Anti Salmonella was highly effective. 27 S. typhimurium strains were isolated from 762 naturally infected chicken giblets by use of unsupplemented Tetrathionate broth. However, 33 S. typhimurium isolates were obtained with ferrioxamine E supplemented Tetrathionate broth from the same samples. Three Salmonella isolates out of 50 evenly divided meat meal samples were obtained by use of ferrioxamine E supplemented buffered peptone water followed by direct streaking onto XLD- and Rambach agars, no Salmonella isolates could be detected by the conventional method. PMID- 8722190 TI - A rapid mechanical lysing procedure for routine analysis of plasmids from lactobacilli, isolated from sourdoughs. AB - Variations in cell wall composition of lactobacilli complicate attempts to standardize plasmid detection on the basis of enzymatic lysing procedures. Mechanical disruption of cell walls by a horizontal mixer-mill proved to be more effective, rapid and reproducible. In order to judge the risk of damaging plasmid deoxyribonucleic acids (DNA) in this way, plasmid profiles obtained after enzymatic lysing and mechanical disruption of cell walls were compared. As no differences were detected the mechanical procedure was accepted as a suitable method for routine analysis of plasmid profiles. PMID- 8722191 TI - Characterization of Lactobacillus acidophilus strains for use as dietary adjunct. AB - Seven Lactobacillus acidophilus strains were studied for their morphology, biochemical properties, growth behaviour, bile tolerance, cholesterol uptake and survival at low pH. The strains significantly differed in their biochemical and beneficial traits. The strains which exhibited cholesterol uptake and ability to grow in the presence of 0.3% bile salt also utilized mannitol. PMID- 8722192 TI - Prevalence and serovar distribution of Salmonella in fresh and frozen meat from captive Nile crocodiles (Crocodylus niloticus). AB - Salmonella was isolated from 6 of 20 (30.0%) samples of fresh meat, and from 28 of 140 (20.0%) samples of frozen meat processed for human consumption from captive Nile crocodiles (Crocodylus niloticus) in Zimbabwe. Salmonella enterica isolates showed a serovar distribution of 41.2% (14/34) subsp. enterica, 11.8% (4/34) subsp. salamae and 41.2% (14/34) subsp. diarizonae. Analyses of fresh meat samples yielded Aeromonas (A. hydrophila) in 18 of 20 samples (90%), and a mean aerobic plate count (APC, 30 degrees C) of 5.79 cfu/g, a mean coliform count (TC, 37 degrees C) of 5.08 cfu/g and a mean faecal coliform count FC, 44 degrees C) of 4.76 cfu/g. It is suggested that the presence of Salmonella in meat samples may be due to skin surface contamination originating from faecally polluted rearing water ponds combined with excessive handling procedures during flaying. The common presence of Salmonella, including serovars of proven pathogenic potential, in crocodile meat offered for human consumption should concern consumers and public health authorities, as well as staff employed at crocodile farms. PMID- 8722193 TI - The stability of stored gari. AB - Gari is a widely consumed stored staple from cassava (Manihot esculenta Crantz). Processing involves peeling, grating, fermentation, dewatering and roasting, resulting in a granular product. It is a shelf stable food consumed as processed or cooked. When retailed, it is subject to post-process contamination. Investigations were carried out to establish the stability of processed gari. Samples of various water activities (a(w)) were packaged in 250 gauge polyethylene bags with or without vacuum packaging and in nylon sacks. Initial and weekly determination of pH, a(w) and mycoflora were undertaken over 16 weeks. An initial pH of 4.3 +/- 0.1 was stable during the experiment. Nylon sack samples with initial a(w) of 0.40 and 0.55 showed an increase of a(w) to 0.65 at the end of the storage. Vacuum packed polyethylene bag samples with initial a(w) of 0.70, 0.55 and 0.40 recorded 30, 20 and 10%, respectively of the initial mycoflora after 16 weeks. A low mycological load, a(w) < 0.70 and vacuum sealed plastic packaging ensure physico-chemical and microbiological stability of stored gari. PMID- 8722194 TI - Cellular and molecular bases of B-cell clonal expansions. AB - The concept that lymphomagenesis is a multistep process is now widely accepted. Various factors are involved in the development and malignant progression of B cell lymphoproliferative disorders. The most frequently recognized alterations in these disorders are chromosomal translocations which lead to the activation of proto-oncogenes (c-myc) or genes encoding for proteins involved in the control of the cell cycle (cyclin D1), differentiation (bcl-6) and apoptosis (bcl-2). In addition, genetic changes that inactivate tumor suppressor genes (p53, Rb, p16) have recently been identified. Infectious agents may also play a role in lymphomagenesis either by directly driving B-cell proliferation (EBV) or by inducing a chronic antigenic stimulation (EBV, HCV, HBV, helicobacter pylori). Finally, several data indicate that local cytokine networks and, in particular, autocrine (IL-6, IL-10) and/or paracrine (IL-2, IL-4, IL-6) loops probably play a contributory role in the development and evolution of B-cell lymphoproliferation. In the last few years, the advent of molecular biology techniques has allowed important advances in the definition of the events involved i the earlier phases of lymphoma development. This has been made possible, in particular, by the study of a series of oligoclonal or monoclonal lymphoproliferative disorders characterized by an indolent or "smoldering" clinical course, such as follicular lymphoma and the lymphoproliferation associated with autoimmune diseases, which are at high risk of evolution to a highly malignant lymphoma. In nearly all of these conditions, the clonal B-cells responsible for the early stages of the disease are probably not fully transformed and retain various degrees of responsiveness to a wide variety of microenvironmental stimuli (antigen or autoantigen stimulation, interactions with "reactive" T lymphocytes, local cytokine networks). These latter in turn may induce the regression of pathological lesions, maintain the disease in an active state or contribute to the evolution towards an overtly malignant lymphoma. These findings open new avenues for the design of unconventional strategies of intervention aimed at preventing the malignant evolution of pre-lymphomatous lesions and controlling the clinical course of certain low-grade B-cell lymphomas. PMID- 8722195 TI - The atypical lymphoproliferative disorders: tissue expression. AB - The atypical lymphoproliferative disorders (ALDs) represent one of the most intriguing problems in diagnostic histopathology. In fact, there is no sharp distinction between benign and malignant lesions within this group of diseases and the name ALD reflects this doubt. New developed molecular and genetic techniques probably will help to solve this diagnostic problem. PMID- 8722196 TI - The role of molecular analyses of B-cell and T-cell clonality in the study of B cell lymphomagenesis. AB - While the simple detection of B-cell clonality does not imply B-cell malignancy, comprehensive analyses of B-cell clonality are crucial to investigate the pathobiology of B-cell lymphoproliferative disorders. Our recent studies in patients with Sjogren's syndrome have highlighted how multiple molecular analyses of B-cell clonality (Southern blot, polymerase chain reaction, single strand conformation polymorphism, and DNA sequence analysis) in prelymphomatous lesions may be of great value in helping to define the stages of progression towards low grade malignancy. The study of T-cell expansion may also be important in investigations of the pathobiology of the different stages of B-cell lymphoproliferation (fully benign, pseudolymphomatous, or definitely malignant), which may still be T-cell-and antigen/ autoantigen-dependent. PMID- 8722197 TI - Monitoring clonal expansion in the laboratory. AB - The definitive demonstration of the clonal origin of an expanded B or T cell population is achieved by the molecular analysis of B or T cell receptor gene rearrangement. However, in recent years techniques that are both sensitive and relatively easy to carry out have been introduced as the first step to the diagnosis of clonality. These include immunochemical analysis of serum and urine to characterize their monoclonal immunoglobulin components, and immunophenotypic studies of peripheral blood or tissue cells. The most widely used techniques are reviewed here, with particular emphasis on the clinical applications of immunofixation electrophoresis and multiparametric cytofluorimetric procedures. PMID- 8722198 TI - Monoclonal gammopathies antigen driven in autoimmune diseases. AB - An important link exists between cancer, autoimmune diseases and immune deficiency. This presentation will review some data concerning clonal growth in B lymphocytes, with focus on the occurrence of autoantibody f2p4vity in the monoclonal proteins, which are the products of the clonally expanded lymphocyte plasma cell population. The role of antigen in causing or promoting monoclonal B cell growth is considered for several different forms of monoclonal gammopathies benign, malignant, and secondary. There is recent demonstration of M-proteins and clonal growth complicating HIV infections and some autoimmune conditions which illustrate well the link between cancer, autoimmunity and immune deficiency. PMID- 8722199 TI - Hepatitis C virus infection and clonal B-cell expansion. AB - The striking association between hepatitis C virus (HCV) infection and the so called "essential" mixed cryoglobulinemia (MC) has led to the hypothesis that HCV plays a major role in the production of cryoglobulins. Analysis of soluble and cryoprecipitable immune complexes shows that the hepatitis C virion is bound to IgM bearing the WA cross-idiotype (XId). The production of WA XId IgM seems to be the result of chronic stimulation by HCV of a population of WA XId + CD5 + B cells. It is possible that the reactivity of WA XId IgM does not initially include rheumatoid factor (RF) activity, which may be acquired secondarily from mutational events accompanying a probably T-cell independent B cell proliferation. Type II MC is a benign proliferation that progresses to malignancy in a minority of patients. This is consistent with the concept that malignancy progression involves the accumulation of multiple mutations of proto-oncogenes and tumor suppressor genes that are facilitated by chronic antigenic stimulation. The recent demonstration of HCV in hyperplastic reactive lymphoadenopathy and in the neoplastic lymph nodes of patients with MC strengthens the putative role played by HCV in lymphomagenesis. A fuller understanding of the virus-related mechanisms of lymphoproliferation in MC patients would contribute significantly to the development of therapeutic strategies. PMID- 8722200 TI - Helicobacter pylori, gastric MALT and B-cell clonality. AB - MALT, or mucosa associated lymphoid tissue, is normally not present in gastric tissue. Its presence is often associated with persistent antigenic stimulation. MALT is a precursor of gastric MALT lymphoma, a low-grade lymphoma whose incidence recently appears to have increased. Although much epidemiologic and clinical evidence has linked both MALT and MALT lymphoma to Helicobacter pylori infection, it is not known whether other agents and or mechanisms may also play a role and whether there is a clearly defined pre-neoplastic lesion. In particular, the clinical significance of B-cell clonality remains unknown. In a recent study we attempted to define the role of H. pylori and MALT in the genesis of B-cell clonality in a northern Italian patient population referred to us for simple dyspepsia. The results show that B-cell clonality is unexpectedly frequent in these patients regardless of the presence of H. pylori infection. These observations raise the possibility that agents and mechanisms other than H. pylori may be involved in the genesis of MALT lymphoma. Indeed, other studies conducted by our group in patients with Sjogren's syndrome indicate that genetic/immunologic factors and possibly viruses may play a role. The high prevalence of B-cell clonality in an otherwise healthy population suggests either that most of these patients are at risk of developing MALT lymphoma (in which case this condition at the moment may be greatly underdiagnosed) or that B-cell clonality is a very early step in the development of neoplasia, which requires several other factors and which will occur only in a restricted fraction of these patients. Careful follow-up studies will provide an answer to this question. PMID- 8722201 TI - Autoimmune diseases and monoclonal gammopathies. AB - Rheumatoid arthritis, systemic lupus erythematosus and primary Sjogren's syndrome are nonorgan-specific autoimmune diseases in which serum monoclonal immunoglobulins (MIg) have been identified repeatedly. Conversely, autoimmune traits have been detected in a number of patients with lymphoproliferative disorders such as multiple myeloma, Waldenstrom's macroglobulinemia and chronic lymphocytic leukemia. The latter cells have even shown to produce multispecific autoantibodies. One connection between connective tissue diseases and Iymphoid malignancies might be established by a limitedfraction of B Iymphocytes expressing the CD5 marker. PMID- 8722202 TI - Sjogren's syndrome, mixed cryoglobulinaemia and the monoclonal gammopathies. AB - Sjogren's syndrome (SS) represents a pathological model of the evolution from polyclonal B lymphocyte activation to oligo-monoclonal B cell expansion, which may culminate in the development of a true lymphoproliferative disease. The different phases of this process are usually marked by the appearance of type II mixed cryoglobulins in which the monoclonal component is commonly represented by an IgM with rheumatoid factor activity. A similar mechanism exists in mixed cryoglobulinaemia (MC), a different pathological entity in which hepatitis C virus infection has been demonstrated to play an important etiopathogenetic role. Although there are significant differences between SS and MC, they also share many clinical and immunological characteristics, which suggest that common pathogenetic mechanisms may underlie both disorders. PMID- 8722203 TI - B-cell lymphoproliferation in primary Sjogren's syndrome. AB - Sjogren's syndrome (SS) is characterized by lymphocytic infiltration of the exocrine glands, along with a polyclonal B-cell activation which is illustrated by the presence of multiple autoantibodies against organ and non-organ specific autoantigens. Lymphoproliferative disorders present with a higher frequency in patients with Sjogren's syndrome. The spectrum of lymphoproliferation extends from an increased frequency of circulating monoclonal immunoglobulins, free light chains and mixed monoclonal cryoglobulins (type II cryoglobulinemia) to an increased frequency of non-Hodgkin's lymphoma. In fact, patients with Sjogren's syndrome have a 44-times higher risk of developing lymphoma compared to the normal population. Several studies have shown that the affected exocrine glands are the major site of monoclonal B-cell proliferation in SS. This has been demonstrated by both immunophenotyping (an increased proportion of kappa:lambda light chains in the B-cells infiltrating the salivary glands) and immunogenotyping studies (monoclonal or oligoclonal light chain gene rearrangements in the salivary glands). The monoclonal rheumatoid factors in the sera of patients with SS share common cross-reactive idiotypes. The presence of these cross-reactive idiotypes is correlated with a higher frequency of extraglandular manifestations, circulating monoclonal immunoglobulins and autoantibodies. The lymphomas, which appear, may vary in type and location; immunocytomas, and intermediate grade and high grade non-Hodgkin's lymphomas have been described. Predictive factors for the development of lymphoma in SS include clinical signs such as lymphadenopathy, splenomegaly, and parotid gland enlargement, as well as laboratory parameters such as the presence of mixed monoclonal cryoglobulinemia and the presence of immunoglobulins bearing the cross reactive idiotypes 17109 (Vk IIIb-related) and G-6 (VH1-related). PMID- 8722204 TI - Autoimmune connective tissue disease, chronic polyarthritides and B cell expansion: risks and perspectives with immunosuppressive drugs. AB - Several autoimmune diseases, including Sjogren's syndrome (SS), systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA), are characterized by B cell hyperactivity, polyclonal activation, and autoantibody synthesis. Overt B cell clonal expansion occurs in a minority of the patients, while at the tissue level clonotypic B cells may be more easily detected in the majority of patients. The data available suggests that antigen-driven B cell expansion, eventually leading to somatic mutation and transformation, is the main event. Immunosuppressive drugs known to increase chromosomal damage and to lead to earlier transformation should therefore be avoided, unless strictly necessary to preserve vital organ functioning. New immunosuppressive drugs such as methotrexate, cyclosporine A, and Rapamycin are promising for they seem to offer effective control of disease related organ damage with acceptable side effects. The B cell lymphoproliferative diseases occurring under treatment seem to remit spontaneously after prompt drug withdrawal. Close surveillance, employing new techniques capable of detecting early B or T cell clonal expansion, may allow better monitoring of possible complications. Biological agents such as alpha-interferon and monoclonal antibodies (which are directed against specific immunological mediators and thus target-selected steps of the immune-inflammatory process) have opened promising new research topics in all these diseases. PMID- 8722205 TI - Public health social work: pro bono publico. PMID- 8722206 TI - Curanderismo: demystifying the health beliefs and practices of elderly Mexican Americans. AB - In Mexican American communities traditional folk healing historically has played an important role in meeting critical health care needs of residents. Despite this fact, researchers who study health care rarely consider the role of folk healing among elderly Mexican Americans, and thus little knowledge is available about its use by this group. In this article, 25 Mexican American elderly people participated in extensive ethnographic interviews about folk healing (curanderismo) and its influence on health care behaviors. Although participants relied on modern medicine to treat serious injuries and major health problems, they still considered traditional folk healing a viable alternative in situations in which modern health care was unsatisfactory or ineffective. Therefore, social workers need to become more knowledgeable about and sensitive to folk beliefs and practices and to develop culturally appropriate health care interventions. PMID- 8722207 TI - Multiple perceptions of discharge planning in one urban hospital. AB - Since the advent of diagnosis-related groups (DRGs), advocacy groups have claimed that although hospital discharge planners perceive the discharge planning process as helpful, elderly patients and their families do not. This article explores how the discharge planning process was perceived by 40 discharge planners and 40 family caregivers. Planners greatly overrated caregiver influence and the amount adequacy of information shared about posthospital health care, choice of discharge to home or nursing home, and time to decide. Caregivers perceived that nursing homes were forced on patients by social workers and physicians. DRGs, physicians, and hospital administrators appeared to pressure social workers to coerce mentally competent patients into nursing homes. Excessive concern by hospital staff about patient safety after discharge may override patients' rights to autonomy and self-determination, violating the NASW Code of Ethics. Implications for practice, policy, and future research are discussed. PMID- 8722208 TI - Provider and consumer profiles of traditional and high-tech home health care: the issue of differential access. AB - This study, based on data from 154 home health care agencies, examines the comparative profiles of traditional and technology-enhanced, or "high-tech", home health care service providers and recipients. Voluntary, not-for-profit agencies have been providing services the longest, serve more patients, make more home visits, and have more staff. However, proprietary programs have grown faster than agencies under other auspices. One in 10 patients receives high-tech care, with hospice agencies serving the largest proportion of high-tech patients. High-tech patients are more likely to be younger, male, married, white, and living with others than other service recipients in general. A slight majority of agencies have special admissions criteria for high-tech patients. Variations in the profiles of traditional and high-tech patients are considered in terms of their implications for differential access to particular categories of in-home services. PMID- 8722209 TI - Applying the codependency model to a group for families of obsessive-compulsive people. AB - Obsessive-compulsive disorder places formidable burdens on family members who are drawn into bizarre enabling behaviors to pacify their ill relatives. Some families exhibit a high tolerance for inappropriate behavior and evidence characteristics similar to those seen in families of alcoholics. This article applies the codependency group model to families of obsessive-compulsive people based on a definition of codependency that views these families as normal, feeling people who are trying to cope with unremitting stress. Clinical vignettes illustrate how these families are similar to families of alcoholics in their management of emotions and in their dysfunctional behaviors. Recommendations are offered for practitioners who work with families of mentally ill people. PMID- 8722210 TI - Interdisciplinary expectations of the medical social worker in the hospital setting: Part 2. AB - This second part of a survey report focuses on the views of physicians, nurses, and social workers concerning the role of medical social workers in addressing health-related patient problems. All three groups, but particularly physicians and nurses, were more likely to see counseling services as a social worker's job when the client was a family member of the patient, rather than the patient, and when the objective was to modify social-environmental problems of the client rather than to support or modify the person. Also, all three groups, but particularly physicians and nurses, were likely to perceive environmental problems of patients, more than emotional or behavioral ones, as an area distinctive of social work. In contrast to previous study findings, physicians and nurses did not wish to exclude social workers in hospitals from counseling patients and from addressing psychosocial problems; they just did not see these activities as distinctive of social work. PMID- 8722211 TI - Social work perspectives in organ procurement. AB - The gap between the supply of and demand for organs for transplantation has widened in the past two decades, resulting in low quality of life and increased mortality for people waiting for a transplant. Current strategies for organ procurement are inadequate; therefore, alternative methods have been suggested. In the center of the debate are the required request strategy, which endorses the informed consent principle, and the presumed consent strategy, which assumes but does not require explicit consent from the donor. To date social workers have not been extensively involved in the debate, although organ procurement issues involve social work values such as individual rights, free choice, and self determination. To become active and influential in the policy of organ procurement, social workers should become familiar with these issues and use crisis intervention techniques with grieving families to help them with the donation decision. PMID- 8722212 TI - Does apologizing help? The role of self-blame and making amends in recovery from bereavement. AB - This article presents the results of an investigation into the relationship between attributions of self-blame for the death of a loved one and subsequent psychological recovery from this loss. Two hundred and forty-four people who had suffered the death of a loved one identified the emotional, cognitive, and behavioral responses they used at the time of their loss. Data indicated that use of self-blame was associated with poorer long-term adjustment. However, it was also found that self-blame often led people to make amends or reparations. Significantly, when self-blame was linked to making amends, it became correlated with favorable adjustment outcomes. The potential therapeutic value of reviewing the moral and spiritual beliefs of self-blaming clients and determining how, within their belief system, they might make amends is considered. PMID- 8722213 TI - Of technological targets and budgetary bombs: the dangers of depersonalized budgetary warfare. PMID- 8722214 TI - Excess of rare cancers in Werner syndrome (adult progeria). AB - The association between genetic disorders and diverse cancers has provided clues for laboratory research into carcinogenesis. Such an opportunity now arises from studies of cancer in Werner syndrome (WRN). Soft-tissue sarcoma (STS) and benign meningioma have been associated with WRN, an autosomal recessive disorder characterized by premature aging, more commonly reported in Japan than elsewhere, in part because of inbreeding. In the literature we found 124 case-reports of neoplasia and WRN from Japan and 34 from outside Japan, 1939-August, 1995. They reveal a greater diversity of neoplasia in WRN than was previously known. In Japanese, there were 127 cancers, 14 benign meningioma, and 5 myeloid disorders, as compared with 30, 7 and 2 respectively in non-Japanese. The ratio of epithelial to non-epithelial cancers was about 1:1 for Japanese and for non Japanese instead of the usual 10:1. Both series had excess of STS, osteosarcoma, myeloid disorders, and benign meningioma. In addition, the Japanese had an excess of thyroid cancer (20 versus 2 cases in non-Japanese) and melanoma (21 versus 3 cases), including 5 intranasal and 13 of the feet. STS, osteosarcoma, melanoma, and thyroid carcinoma accounted for 57% of all cancer in WRN as compared with 2% expected based on the Osaka population at 25-64 years of age. Multiple tumors were reported in 19 Japanese and 5 non-Japanese. In Japan, nine first-degree relatives had WRN and cancer, six of whom were concordant as to site and/or cell type. The WRN gene has been mapped to chromosome 8p. The high frequency of thyroid cancer and melanoma in Japanese, not found in Caucasians, may be related to a report of linkage disequilibrium with the WRN gene in Japanese but not in Caucasians and to haplotype differences within and between the two races, suggesting multiple independent mutations. PMID- 8722215 TI - Ethnic variation in prostate cancer survival in New Mexico. AB - Prostate cancer survival varies markedly by ethnicity. American Indians and blacks have the lowest 5-year relative survival among ethnic groups in the U.S. In New Mexico, relative survival for prostate cancer is lower for ethnic minority groups than for non-Hispanic whites. To examine factors underlying ethnic differences in prostate cancer survival in New Mexico, we analyzed Surveillance, Epidemiology, and End Results Program data collected by the New Mexico Tumor Registry from 1983 to 1992. Unadjusted relative risk (RR) of death after prostate cancer diagnosis was greater for Hispanics [RR = 1.1; 95% confidence interval (CI), 1.0, 1.2], American Indians (RR = 1.4; 95% CI, 1.2, 1.5), and blacks (RR = 1.5; 95% CI, 1.2, 1.7) than for non-Hispanic whites. After adjusting for age, stage, histological grade, year of diagnosis, and initial treatment, the risk for Hispanics (RR = 1.0; 95% CI, 0.9, 1.1), American Indians (RR = 1.0; 95% CI, 0.9, 1.1), and non-Hispanic whites was comparable. Although based on small numbers, adjusted risk ratios among blacks remained elevated (RR = 1.2; 95% CI, 0.9, 1.6), due in part to lower survival during the first 12 months after diagnosis (RR = 2.0; 95% CI, 1.2, 3.3) and poorer survival following radical prostatectomy (RR = 4.2; 95% CI, 1.3, 13). These findings suggest that poorer survival for Hispanics and American Indians may be explained by delayed detection and differences in treatment. PMID- 8722216 TI - Temporal patterns of aflatoxin-albumin adducts in hepatitis B surface antigen positive and antigen-negative residents of Daxin, Qidong County, People's Republic of China. AB - Molecular epidemiological studies of populations at high risk for liver cancer have shown that hepatitis B virus (HBV) and aflatoxin B1 (AFB1) exposure are two major risk factors for this disease. These etiological agents, combined with nutritional deficiencies, are important for the initiation and promotion of liver cancer in various parts of the world. In Qidong, People's Republic of China, liver cancer accounts for 10% of all adult deaths, and both HBV and AFB1 exposures are common. To study temporal and possible chemical-viral interactions in people, serum samples were collected during a longitudinal study designed to measure aflatoxin molecular biomarkers in residents of Daxin Township, Qidong City, People's Republic of China. In this study, the temporal modulation of aflatoxin adduct formation with albumin over multiple lifetimes of serum albumin was examined in both HBV-positive and HBV-negative people in two periods: September-December 1993 (wave 1) and June-September 1994 (wave 2). During the 12 week monitoring period of wave 1, 120 individuals (balanced by gender and HBV status) provided a total of 792 blood samples. AFB1-albumin adducts were detected in all but one of the serum samples. The range of binding detected by RIA in the Daxin population was 0.17-4.39 pmol AFB11/mg albumin with an overall mean +/- SD of 1.51 +/- 0.21 pmol AFB11/mg albumin. The mean +/- SD for weeks 0, 2, 4, 6, 8, 10 and 12 of wave 1 were 1.21 +/- 0.41, 1.58 +/- 0.70, 1.36 +/- 0.52, 1.71 +/- 0.44, 1.18 +/- 0.60, 2.00 +/- 0.59, and 1.68 +/- 0.34 pmol AFB1/mg albumin, respectively. During wave 2, 103 individuals from wave 1 provided a total of 396 blood samples collected monthly over wave 2, with mean +/- SD aflatoxin-albumin adduct levels of 1.19 +/- 0.37, 0.85 +/- 0.45, 0.89 +/- 0.28, and 0.61 +/- 0.15 pmol AFB1/mg albumin. Using linear regression models, the mean aflatoxin-albumin adduct levels increased (P < 0.05) during the 12 weeks of wave 1 and decreased (P < 0.05) over the 4 months of wave 2. Neither HBV surface antigen status nor gender modified either the baseline mean or the temporal trend. High-performance liquid chromatography confirmation was done on a subset of serum samples, and the results show an excellent association between the immunoassay data and high performance liquid chromatography. Taken together, these data demonstrate that AFB1-albumin is a sensitive and specific biomarker for assessing exposure to this carcinogen in the population in Qidong. PMID- 8722217 TI - Effects of dietary supplementation of alpha-tocopherol on plasma glutathione and DNA repair activities. AB - In a randomized double-blind trial of alpha-tocopherol (vitamin E), we investigated the effects of alpha-tocopherol supplementation on lipid- and water soluble antioxidants in plasma and DNA repair activities in peripheral mononuclear leukocytes. Baseline levels of antioxidants and DNA repair activities were assessed twice before alpha-tocopherol intervention: on day 1 (visit 1) and day 3 (visit 2). During the second visit, participants were randomized to receive one of three dosages of alpha-tocopherol, 15, 60, or 200 mg/day for 4 weeks. The same biochemical measurements as at baseline were repeated twice after intervention: on day 17 (visit 3) and day 31 (visit 4). A total of 31 healthy volunteers were eligible for the study, completed all four visits and were included in the final data analysis. At baseline, no appreciable differences of dietary intake of vitamin E and plasma alpha-tocopherol were observed among the three dosage groups. In general, supplementation of alpha-tocopherol for 2-4 weeks resulted in a dose-dependent increase of plasma level of alpha-tocopherol (compared to baseline); significant increases of plasma alpha-tocopherol at visits 3 and 4 were observed in the two higher dosage groups, 60 and 200 mg, but not in the lowest dosage group, 15 mg. At visit 4 (but not visit 3), plasma glutathione levels were significantly elevated (compared to baseline) in the two higher dosage groups, 60 and 200 mg, but not in the lowest dosage group, 15 mg. In addition, there was an increase in the lipid protection ratio by supplementation of alpha-tocopherol for 2-4 weeks in the two higher dosage groups, 60 and 200 mg, but not in the lowest dosage group, 15 mg. In general, there were no consistent effects of alpha-tocopherol on DNA repair activities in peripheral mononuclear leukocytes after being adjusted for baseline DNA repair activities. Results from this study demonstrate the interrelationship between alpha-tocopherol and other antioxidants in plasma; total plasma antioxidants can be modulated by short-term dietary supplementation of alpha-tocopherol. PMID- 8722218 TI - Human papillomavirus DNA in invasive cervical carcinomas and its association with patient survival: a nested case-control study. AB - This study sought to examine the association between the presence of human papillomavirus (HPV) DNA in invasive cervical cancer and prognosis. A case control study was undertaken nested in a cohort of 208 patients with invasive cervical carcinoma in Montreal. All 40 deceased patients formed the case groups. A control group of equal size was selected by matching to each case (1:1) a patient of the same age and year of admission who had survived her disease. HPV DNA was detected by the use of a PCR protocol. The odds ratio (OR) for cervical cancer death was computed by logistic regression. Detection of HPV, particularly of types 16 and 18, was negatively correlated with disease stage and histological grade. The OR for death was 0.27 [95% confidence interval (CI), 0.1-0.8] for those whose tumors were positive for HPV DNA versus those in whom HPV DNA was not detected. After adjusting for the confounding effects of stage and grade, the prognostic effect was somewhat reduced, with an OR of 0.34 (CI, 0.1-1.1), which was no longer significant. The magnitude of the HPV-survival association was not altered when the analyses were restricted to carcinomas of stages I and II. Regardless of the mechanism for the prognostic role, detection of HPV DNA in primary tumors may play an important adjunct role in assessing prognosis of patients with invasive cervical cancer. PMID- 8722219 TI - p53 mutational spectrum of esophageal carcinomas from five different geographical locales in China. AB - A mutational spectrum for exons 5-8 of the p53 tumor suppressor gene in esophageal carcinomas in mainland China and Hong Kong was established. This study involved 209 squamous cell carcinoma specimens obtained from five different geographical locales in China: Zhengzhou, Taiyuan, Shantou, Guangzhou, and Hong Kong. Zhengzhou and Shantou were high-incidence regions for esophageal cancer, whereas the other three regions had low or intermediate incidence of the disease. Analysis by single-strand conformation polymorphism and DNA sequencing showed that 87 specimens (41.6%) contained mutations in exons 5-8 of the p53 gene compared to 163 cases (78%) that had accumulation or aberrant expression of the protein, as detected by immunohistochemical staining. Point mutations accounted for 80.4% (87/107) of all genetic changes. The specimens from northern China exhibited fewer p53 gene aberrations and a more even distribution of mutations in exons 5-8 compared to those from southern China in which 60% of all mutations were found in exon 5. A major hot spot was found at codon 176 in exon 5, where 41 samples from Shantou, Guangzhou, and Hong Kong had a G-->T transversion. It is likely that among southern Chinese this codon is susceptible to mutagenesis by carcinogens. Codons 175, 203, 245, 250, 273, and 282 were also shown to be mutational hot spots, with three or more mutations observed at each site. The p53 mutational data obtained in this study showed that Chinese esophageal carcinomas are often associated with some unique genetic alterations, which may be attributed to specific dietary or environmental carcinogens that affect the Chinese but not Caucasians. PMID- 8722220 TI - Familial and nutritional risk factors for p53 overexpression in colorectal cancer. AB - Although familial and dietary factors are recognized as important risk determinants for colorectal tumorigenesis, the specific causes of colorectal cancer remain unclear. Studies of p53 genetic alterations have provided clues concerning the etiology of many cancers. This study was designed to determine whether overexpression of the p53 protein is associated with familial and dietary risk factors. Epidemiological data were obtained from 163 colorectal cancer cases and 326 healthy controls. Tumors of all patients were analyzed immunohistochemically for p53 overexpression using an avidin-biotin immunoperoxidase procedure and polyclonal anti-p53 antibody CM1. Of patient tumors, 44.8% showed p53 nuclear reactivity. Colorectal cases versus controls were three times more likely to report a family history of colorectal cancer [odds ratio (OR), 3.12; 95% confidence interval (CI), 1.77-5.52]. Only cruciferous vegetables exhibited a significant inverse association (OR, 0.59; 95% CI, 0.34-1.02; trend test, P = 0.03) for the highest versus lowest quartiles. Both meat and beef displayed an elevated increase in risk. When cases with p53 overexpression (p53 positive) were compared with cases without p53 overexpression (p53 negative), etiological heterogeneity was suggested for family history of colorectal cancer (OR, 0.39; 95% CI, 0.16-0.93), cruciferous vegetables (trend test, P = 0.12), and beef consumption (trend test, P = 0.08). To estimate the individual relative risks for p53-dependent and p53-independent pathways, each p53 subtype was compared with controls. Cruciferous vegetables exhibited a significant association (OR, 0.37; 95% CI, 0.17-0.82; trend test, P = 0.03) when p53 positive cases were compared with controls. When p53 negative cases were compared with controls, a significant increase in risk was observed for family history of cancer (OR, 4.46; 95% CI, 2.36-8.43) and beef (OR, 3.17; 95% CI, 1.83 11.28; trend test, P = 0.006). The p53 (positive) dependent pathway was characterized by an inverse association with cruciferous vegetables, and p53 independent tumors were characterized by family history and beef consumption. These data may indicate the significance of linking epidemiology and molecular biology in assessing specific etiological pathways. PMID- 8722221 TI - Detection of tyrosinase mRNA from the blood of melanoma patients. AB - Surgical therapy for localized melanoma is highly successful. However, if melanoma spreads beyond its primary site, the results of treatment are poor. Therefore, early detection of circulating melanoma cells in the blood may be important. Currently, circulating melanoma cells are undetectable. Tyrosinase is an enzyme in the melanin synthetic pathway the expression of which is only found in melanin-producing cells. Because melanocytes are not normally found in the peripheral blood, we hypothesize that melanoma cells circulating in the peripheral blood could be detected by amplifying the tyrosinase mRNA using the reverse transcription-PCR (RT-PCR). The purpose of this study was to determine the sensitivity of a RT-PCR-based assay for tyrosinase mRNA from peripheral blood and evaluate correlations with tumor status in melanoma patients. RNA was isolated from the peripheral blood or tissue culture cells, and cDNA was prepared. DNA was amplified using RT-PCR with nested primers for tyrosinase and beta(2)-microglobulin. Serial dilution experiments using cells from the SK-MEL-28 cell line were performed in culture media and in whole blood. Twelve patients with melanoma, 10 healthy controls, and 15 patients with nonmelanoma malignancies were tested for tyrosinase expression in peripheral blood. The sensitivity of this assay was determined to be as low as 1 melanoma cell in 5 ml of whole blood. No tyrosinase was found in healthy subjects or other cancer control patients. Tyrosinase mRNA was detected in the blood of five melanoma patients (one stage II, two stage III, and two stage IV). Three of these tyrosinase-positive patients had biopsy-proven evidence of melanoma, whereas the other two had no clinical evidence of malignant disease after surgical resection. The remaining seven melanoma patients had no evidence of disease and tested negative for tyrosinase mRNA. This study suggests that a RT-PCR-based assay for the detection of tyrosinase mRNA in peripheral blood is feasible. Moreover, the presence of tyrosinase mRNA in the blood seems to correlate with the stage of melanoma. Further study and follow-up are needed to clarify the role of tyrosinase mRNA as a tumor marker for malignant melanoma. PMID- 8722222 TI - Human metallothionein gene expression determined by quantitative reverse transcription-polymerase chain reaction as a biomarker of cadmium exposure. AB - Expression of the metallothionein (MT) gene in frozen human lymphocytes has been developed as a new molecular biomarker of heavy metal exposure. Workers at a Polish battery factory with high exposure to cadmium were monitored for airborne exposure and blood cadmium levels. A novel quantitative reverse transcription-PCR (RT-PCR) technique, making use of a homologous internal standard, was used to assess the level of MT-specific mRNA in frozen stored aliquots of blood samples taken from exposed and control workers. Results from this assay showed a statistically significant 2.5-fold increase in MT mRNA in exposed compared to control workers. The RT-PCR results also showed significant correlation with airborne cadmium, as registered on personal monitors and with blood cadmium levels. The results suggest that gene induction measured by quantitative RT-PCR is a promising approach for application as a biomarker of biologically effective dose in small samples of frozen tissues or cells. PMID- 8722223 TI - The practice of causal inference in cancer epidemiology. AB - Causal inference is an important link between the practice of cancer epidemiology and effective cancer prevention. Although many papers and epidemiology textbooks have vigorously debated theoretical issues in causal inference, almost no attention has been paid to the issue of how causal inference is practiced. In this paper, we review two series of review papers published between 1985 and 1994 to find answers to the following questions: which studies and prior review papers were cited, which causal criteria were used, and what causal conclusions and public health recommendations ensued. Fourteen published reviews on alcohol and breast cancer and 6 published reviews on vasectomy and prostate cancer were examined. For both series of reviews, nearly all available published studies were cited except for ecological studies and prior reviews. Sources of causal criteria were often not provided. When they appeared, all citations were either the 1964 Surgeon General's report or works of Austin Bradford Hill. Reviews often excluded and sometimes altered criteria without giving reasons for these changes. The criteria of consistency and strength of association were almost always used accompanied by dose-response and biological plausibility in a majority of reviews. The criterion of temporality, considered by many methodologists to be a necessary causal condition, was infrequently used. Confounding and bias were often added considerations. Public health recommendations were not discussed in nearly one-half of the reviews. PMID- 8722224 TI - Recurrent adenomatous polyps and body mass index. AB - Interest in risk factors for the recurrence of adenomatous polyps derives from the use of recurrent adenomas as surrogate end points in longitudinal studies of invasive colorectal cancer. In this case-control study, the effect of increased body mass index (BMI) on the risk of recurrent adenomas was investigated. Subjects consisted of patients seen at three colonoscopy practices in New York City, all of whom had a previous history of adenomas. On index colonoscopy, recurrent cases had an adenoma, whereas controls were normal. Men and women were analyzed separately, with different logistic models developed using backward elimination from a full model containing the covariates age at diagnosis, age-at highest-weight, pack-years of smoking, activity level, energy intake, and fat and fiber intake. Men in the upper quartiles of BMI were found to be at greater risk of recurrent adenomas. In a model which controlled for age at diagnosis, age-at highest-weight, activity level, pack-years of smoking and kilocalories, the estimated odds ratios were 2.2, 1.9 and 1.9 respectively for the second, third and fourth quartiles compared to the first quartile. Only the estimate for the second quartile was found to be statistically significant. No effect was observed for women, even in a model which controlled for age at diagnosis, age-at-highest weight, pack-years and total fat. Obesity may play a role in adenoma recurrence. Confirmation of this finding would have important implications for possible prevention strategies in the future. PMID- 8722225 TI - Regional differences in the incidence and treatment of carcinoma in situ of the breast. AB - Greater use of mammography in the United States in recent years has increased the detection of early neoplasms of the breast, including carcinoma in situ. However, the occurrence and treatment of diagnosed carcinoma in situ of the breast has not been fully described. Our goal was to examine temporal, geographic, and demographic patterns in the incidence and treatment of in situ breast cancer. The study included data from all women with in situ breast cancer that had been detected in the nine Surveillance, Epidemiology, and End Results areas of the United States from 1975 through 1990 (Surveillance Program, Cancer Statistics Branch, Bethesda, MD: National Cancer Institute, November, 1993). We calculated age-adjusted incidence rates (1970 United States standard) using data on histology and treatment from the Surveillance, Epidemiology, and End Results data tape. We assessed predictors of treatment by mastectomy using multiple logistic regression. From 1975-1979 to 1986-1990, the age-adjusted incidence rate of in situ breast cancer increased from 4.7 to 16.9/100,000 women. The increase occurred in all age groups and among both white and black women. However, there was nearly a 2-fold difference in incidence rates across geographic areas in 1986 1990, ranging from < 12/100,000 in Iowa and New Mexico to > 20/100,000 in San Francisco and Seattle. Geographic variability in treatment was also evident, with mastectomy, rather than breast-conserving therapy, performed on 46% of the women with in situ breast cancer in San Francisco and on 66% of those in Iowa. The incidence of diagnosed in situ breast cancer increased markedly during the 1980s, and there was substantial geographic variability in the rates of detection of these tumors and in the type of therapy received. Although mastectomy became a less common treatment over time, it was still performed on a high proportion of women with in situ breast cancer during the latter part of the decade. PMID- 8722226 TI - Experimental chiroptical verification of linkage flexibility in methyl 3-O-(alpha d-mannopyranosyl)-alpha-d-mannopyranoside. AB - Vacuum UV CD spectra of methyl 3-O-(alpha-D-mannopyranosyl)-alpha-D- mannopyranoside in D2O and as a cast film were obtained in the 145-200 nM region. The disaccharide solution CD per residue is nearly identical to that of the monosaccharide solution CD, and to the monosaccharide film CD. Conversely, the disaccharide film spectrum exhibits a strong positive CD linkage contribution in the 160-170 nm range, which is consistent with the known crystal conformation under the aegis of previously determined sector rules. The close similarity between the monosaccharide and disaccharide solution spectra, therefore, reflects conformational averaging in which the net linkage contribution is approximately zero. The present observation of significant solution linkage flexibility confirms previous conclusions based on optical rotation, as well as conclusions of others based on nmr data. Moreover, when combined with those earlier results, the present work demonstrates the population of at least three distinct potential energy wells on the disaccharide phi,psi potential energy surface. PMID- 8722227 TI - Quantum mechanical calculations and experimental measurement of N-terminal charge effects on 1HN and 1HC alpha chemical shifts in peptides. AB - Nuclear magnetic resonance spectroscopists are increasingly utilizing chemical shifts to characterize the secondary structure of proteins. The present study addresses the effects that the positively charged amino group at the N-terminus of a peptide has on 1HN and 1HC alpha chemical shifts along the chain. This information is necessary for interpreting chemical shift data for proteins and/or for peptides that are used as models for protein structure. The chemical shifts for the 1H resonances of four peptides that differ only in the location of their N-terminii are assigned using two-dimensional nmr spectroscopy. The peptides have sequences derived from the beta subunit of the glycoprotein hormone human chorionic gonadotropin (hCG-beta). Comparison of the 1HN and the 1HC alpha chemical shifts for residues common to all four peptides reveals downfield shifts for 1HN and the 1HC alpha resonances within three residues of the N-terminus compared with chemical shifts in the interior of the peptide. The magnitude of the downfield shift is larger for resonances nearer the N-terminus. Quantum mechanical calculations of the 1HN and 1HC alpha chemical shifts in peptides constructed with six alanine units also predict a significant terminus effect. The calculations agree both qualitatively and quantitatively with the experimental data. The inductive nature of the end effect is confirmed in the calculations by Mulliken population analysis. End effects should be taken into account in determining protein secondary structures from chemical shifts. PMID- 8722228 TI - Effects of the addition of hyaluronate segments with different chain lengths on the viscoelasticity of hyaluronic acid solutions. AB - The effects of the addition of sodium hyaluronate segments (sHA) with different chain lengths on the viscoelasticity of high molecular weight sodium hyaluronate (HA) aqueous solutions were studied. The additive effects of sHA depended on the chain length of sHA. Shorter sHA decreased storage (G') and loss (G") moduli, whereas longer sHA increased both moduli. In physiological saline, the effects of the addition of sHA depended on the chain length of sHA, as in aqueous solutions, except that shorter segments did not decrease both moduli. The effects of the addition of monosaccharides, i.e., N-acetyl glucosamine (GlcNAc) and sodium glucuronate (GlcANa), were also studied. Although GlcNAc increased G' and G", GlcANa decreased both moduli. A shift factor "A," which depended on chain-length effects, a sugar effect, and a salt effect, was developed to explain these additive effects quantitatively. PMID- 8722229 TI - DNA structural features responsible for sequence-dependent binding geometries of Hoechst 33258. AB - The complexes of Hoechst 33258 with poly[d(A-T)2], poly[d(I-C)2], and poly[d(G C)2], and poly[d(G-m5C)2] were studied using linear dichroism, CD, and fluorescence spectroscopies. The Hoechst-poly[d(I-C)2] complex, in which there is no guanine amino group protruding in the minor groove, exhibits spectroscopic properties that are very similar to those of the Hoechst-poly[d(A-T)2] complex. When bound to both of these polynucleotides, Hoechst exhibits an average orientation angle of near 45 degrees relative to the DNA helix axis for the long axis polarized low-energy transition, a relatively strong positive induced CD, and a strong increase in fluorescence intensity--leading us to conclude that this molecule also binds in the minor groove of poly[d(I-C)2]. By contrast, when bound to poly[d(G-C)2] and poly[d(G-m5C)2], Hoechst shows a distinctively different behavior. The strongly negative reduced linear dichroism in the ligand absorption region is consistent with a model in which part of the Hoechst chromophore is intercalculated between DNA bases. From the low drug:base ratio onset of excitonic effects in the CD and fluorescence emission spectra, it is inferred that another part of the Hoechst molecule may sit in the major groove of poly[d(G C)2] and poly[d(G-m5C)2] and preferentially stacks into dimers, though this tendency is strongly reduced for the latter polynucleotide. Based on these results, the importance of the interactions of Hoechst with the exocyclic amino group of guanine and the methyl group of cytosine in determining the binding modes are discussed. PMID- 8722230 TI - A fluorescence and CD study on the interaction of synthetic lipophilic hepatitis B virus preS(120-145) peptide analogues with phospholipid vesicles. AB - The interaction of the immunogenic peptide of human hepatitis B virus (HBV) preS(120-145), including B and T epitopes, with phospholipid vesicles has been studied by fluorescence techniques and CD. In addition, interaction of three lipopeptides derived from preS(120-145) containing stearoyl, cholanoyl, and tripalmitoyl-S-glyceryl-cysteine (Pam3C) SS moieties with dipalmitoylphosphatidylcholine (DPPC) has been investigated by polarization fluorescence spectroscopy. Fluorescence experiments showed an increase in fluorescence intensity and a blue shift of the maximum emission wavelength upon interaction of preS(120-145) with DPPC vesicles below the transition temperature (Tc), indicating that the tryptophan moiety enters a more hydrophobic environment. Moreover, fluorescence polarization experiments showed that the peptide decreased the membrane fluidity at the hydrophobic core, increasing the Tc of the lipid and decreasing the amplitude of the change of fluorescence polarization associated with the cooperative melting of 1,6-diphenyl-1,3,5 hexatriene labeled vesicles. The absence of leakage of vesicle-entrapped carboxyfluorescein indicates that the peptide did not promote vesicle lysis. Besides, the three lipopeptides derived from preS(120-145) showed a more pronounced rigidifying effect at the hydrophobic core of the bilayer, with a significative increase in the Tc. Stearoyl- and cholanoyl-preS(120-145) restricted the motion of lipids also at the polar surface, whereas Pam3CSS preS(120-145) did not alter the polar head group order. Finally, CD studies in 2,2,2-trifluoroethanol or in presence of vesicles suggested that the bound peptide adopted amphiphilic alpha-helical and beta-sheet structures, with an important contribution of the beta-turn. It is concluded that preS(120-145) can interact with the lipid membrane through the formation of an amphipathic structure combination of beta-sheet and alpha-helix aligned parallel to the membrane surface, involving the N-terminal residues, and penetrating only a short distance into the hydrophobic core. The C-terminal part, with a combination of beta-turn and beta-sheet structure, remains at the outer part of the bilayer, being potentially accessible to immunocompetent cells. Furthermore, coupling of an hydrophobic moiety to the N-terminal part of the peptide favors anchoring to the membrane, probably facilitating interaction of the peptide with the immunoglobulin receptor. These results are in agreement with the induction of immune response by preS(120-145) and with the enhanced immunogenicity found in general for lipid-conjugated immunopeptides. PMID- 8722231 TI - A pseudo-particle approach for studying protein-ligand models truncated to their active sites. AB - A molecular dynamics method has been developed to describe the structural and dynamic properties of protein-ligand complexes that are truncated to their active sites. The active site is comprised of the ligand and discontinuous, positionally unrestrained peptide chains. This truncated active-site complex is surrounded by big unspecific pseudo-particles representing the complete protein and the solvent. Thus, knowledge of the folding of the outer parts of the protein is not required, and the method can be applied to protein models, derived from homology modeling. The method has been tested using ligand complexes of adenylate kinase, retinol binding protein, HIV-1 protease, and human leucocyte antigen. Comparisons with their crystal structures and with results from time-demanding simulations of the whole complexes in explicit water solvent show that the ligand binding properties are conserved. Most of the hydrogen bonds between the ligand and the active-site residues are reproduced and, furthermore, the simulation time is reduced. PMID- 8722232 TI - Determination of the secondary structure of isomeric forms of human serum albumin by a particular frequency deconvolution procedure applied to Fourier transform IR analysis. AB - A new deconvolution procedure was applied to the analysis of Fourier transform ir spectra of human serum albumin secondary structure in the native state and in states denatured by heat and acid treatment. The deconvolution method is based on the use of the Conjugate Gradient Minimization Algorithm, with the addition of suitable constraints directly obtained by the application to the measured spectrum of the second derivative operator. This method computes central band frequency, bandwidth, and amplitude of the different spectral components of conformation-sensitive amide bands. In the specific case, it was applied to analysis of the amide I band, and the quantitative determination of the different secondary structures (alpha-helix, beta-sheet, beta-turns, and random) was attempted for all the samples examined. The precision of the quantitative determination depends on the amounts of these structures present in the protein. The coefficient of variation is < 10% for values of amide I component > 15%. The accuracy was tested by comparing, by means of linear regression, the results obtained for human serum albumin, hemoglobin, alpha-chymotrypsin, and cytochrome c, using our method, with those obtained by x-ray crystallography and CD; the results obtained by other vibrational spectroscopic approaches were also compared. The fit standard error between x-ray and ir secondary structure values estimated by our method is 2.5% for alpha-helix, 7.16% for beta structures, and 5.1% for other structures (turns and random coils). Quantitative results are given for the secondary structures (alpha-helix, turns, and beta-strands) present in the native state (turns and beta-strands up to now unknown in aqueous solution), together with the percentages of these structures and additional ones (random coils and beta-sheets) formed during denaturization. PMID- 8722233 TI - Tyrosines in two-stranded coiled coils are CD active near 280 nm even in the absence of interhelix tyrosine-tyrosine interactions. PMID- 8722234 TI - Bone densitometry: patients with end-stage renal disease. AB - Bone mass loss and osteoporosis are associated with various conditions, such as end-stage renal disease (ESRD), and treatments, such as prolonged steroid therapy. Bone densitometry is used to measure bone mass density to determine the degree of osteoporosis and to estimate fracture risk. Bone densitometers measure the radiation absorption by the skeleton to determine bone mass of the peripheral, axial, and total skeleton. Common techniques include single-photon absorptiometry (SPA) of the forearm and heel, dual-photon (DPA) and dual-energy x ray absorptiometry (DXA) of the spine and hip, quantitative computed tomography (QCT) of the spine or forearm, and radiographic absorptiometry (RA) of the hand. Part I of this report addresses important technical considerations of bone densitometers, including radiation dose, site selection, and accuracy and precision, as well as cost and charges. Part II evaluates the clinical utility of bone densitometry in the management of patients with ESRD. End-stage renal disease affected more than 242,000 Americans in 1992, and each year 10,000 to 20,000 new cases are diagnosed. Although the survival rate of ESRD patients has improved, metabolic bone diseases that fall under the generic term "renal osteodystrophy" represent abnormal development of bone and major long-term complications. Issues addressed are the type and extent of bone loss associated with ESRD and whether these patients have an increased risk for fracture. The other assessments in this series address the clinical utility of bone densitometry for patients with asymptomatic primary hyperparathyroidism, steroid dependent patients, estrogen-deficient women, and patients with vertebral abnormalities. PMID- 8722235 TI - [Patients' attitudes toward their schizophrenic illness after their hospitalization]. AB - Description of patients' attitudes toward their experience of illness as well as relationships between these attitudes and some other clinical variables (praemorbid background, picture and dynamics of clinical status, insight of illness, and indices of early prognosis) were examined in the group of 20 firstly addmitted inpatients suffering for schizophrenia (ICD-10 criteria). As a measure the scale "Experience of Illness" was used allowing to point the typus of the whole attitude (isolating, undecided, or integrating) and its three dimensions (cognitive, emotional and behavioural). Isolating or undecided attitudes prevailed over assimilating ones when patients were leaving the hospital (2:2:1). In addition, the findings suggest also that assimilating the illness was for patients more easy to achieve in the domain of knowledge about it (identifying the illness with self) or of reaction to it (ability to reflection) than in the domain of feelings (readiness to evaluate it positively). Some interesting correlations between attitudes and clinical variables need confirmation in larger sample of patients. PMID- 8722236 TI - [Patients' attitudes toward their psychosis after its long-term duration]. AB - The aim of the study was to analyze frequency of various types of patients' attitudes toward their illness and correlations between these attitudes and other clinical and social variables. Attitudes of 107 patients suffering from delusional psychoses (70% of them schizophrenic) were studied during follow-up examination ca. 8.7 years after their first hospitalization and 11.2 years after the onset of their illness. The attitudes were assessed by the "Experience of Illness" scale. Its results were correlated with several measures--indices of intensity of hospitalization during the follow-up period, as well as patients' clinical and social functioning at the end of this period. The isolating attitudes predominated over the undecided and integrating (7:4:1). Integrating attitudes correlated with earlier age of the onset of illness, and paranoid structure of delusional syndroms. They also correlated sligthly with better life functioning as well as with lower intensity of residual symptomatology. No relationships between attitudes and nosological position of the illness (schizophrenia versus non-schizophrenia) were stated. The implications of results for future treatment programs is briefly discussed. PMID- 8722237 TI - [The frequency of occurrence of negative symptoms at the early stage of schizophrenic psychosis]. AB - 103 patients were admitted for the first time to the psychiatric hospital (Institute of Psychiatry and Neurology in Warsaw) between 1976-1983 and received a research diagnosis of schizophrenia (in accordance with the ICD-9 criteria). The course and clinical pattern of the illness were analyzed at a follow-up in 65 patients--rehospitalized in the 5th year from their first admission. As regards the clinical pattern analysis, it was focused mostly on positive and negative symptoms occurrence, as assessed using the Andreasen Scales (SANS, SAPS). The data obtained from the case reports were statistically tested and the results were presented in the tables according to the research questions. The frequency of occurrence of at least one negative symptom was found to be high in the group under study (in 72% of patients during the first hospitalization and in 83% at the five-years' follow-up). The increment in the frequency of occurrence pertained also to all groups of negative symptoms, which is concordant with many authors, results. PMID- 8722238 TI - [Frequency analysis of verbal associations obtained from schizophrenics and normals. Formal study]. AB - An association experiment was conducted in a group of schizophrenic people and in a group of sane ones. Words varied as to their emotional shade were given as the stimuli. There were 4 words emotionally neutral, 4 words with a positive emotional shade, 4 words with a negative one and 4 words with an aggressive one. The hierarchies obtained in the group of the insane people had a smaller range: the "frequent" responses were more rare here and the "rare" ones more frequent; the strength of the dominating response was also reduced. All these characteristics were clearer in the relation to the emotionally shaded stimuli, especially the aggressive ones. The views of Broen and Storms as well as Mednick and Osgood were corroborated here. PMID- 8722239 TI - [Schizophrenia combination therapy with neuroleptics and benzodiazepines]. AB - The literature review of combination therapy of schizophrenia with neuroleptics and benzodiazepines is presented. Early trials from the 60s are difficult to evaluate because of the heterogeneity of the patients' groups selected, non standardized evaluation methods and different diagnostic criteria. The more recent trials, methodologically more correct, were aimed at the allevation by the benzodiazepines of anxiety and agitation symptoms often observed in schizophrenia. The trials aimed at the efficacy of co-therapy with neuroleptics and benzodiazepines, (mainly alprazolam) in schizophrenia were less frequent. As the result, not univocally positive effect of such a therapy was found. The differences in clinical effect might be caused by the individual susceptibility to the anxiolytics. The attempts to correlate various clinical effects of combination therapy with neuroleptics and benzodiazepines and the biological changes in central nervous system in schizophrenia, are also discussed. PMID- 8722240 TI - [Successful therapy of the psychiatric patient with catatonic paranoidal syndrome and monitoring of induced cytokine production]. AB - The paper reports the case of a 42-year-old woman with a diagnosed catatonic paranoidal syndrome and a noted dysfunction as regards lymphocyte subpopulation and the production of some cytokins: Tumor Necrosis Factor (TNF) and interferons (IFN). As the first phase of the treatment had proved unsuccessful and there appeared symptoms hinting at the possibility of a subclinical inflammatory process in the brain, parallel to psychiatric therapy, treatment was instituted in the system of successive three-week cycles with Tolpa's Peat Extract (PTT), Selenium and Zinc on a natural yeast basis (Selenium and Zinc preparations) as well as with encortone. After a month of treatment a complete clinical remission was achieved as well as normalization as regards all the tested immunological markers. PMID- 8722241 TI - [Depression in schizophrenia. Comments on possibilities to use fluoxetine]. AB - Depressive symptoms are frequently diagnosed in patients with schizophrenia. The coexistence of mood disorders and criteria of schizophrenia may result in the diagnosis of schizoaffective disorders. Depressive symptoms can also be recognized after the acute phase of schizophrenia or after neuroleptic treatment which results in the diagnosis of postpsychotic or postneuroleptic depression. Sometimes, depressive symptoms can be misinterpreted; it happens because depressive symptoms can resemble negative symptoms of schizophrenia or neuroleptic side effects. The principles of therapeutic approach to depressive symptoms in schizophrenia have been discussed. Taking into consideration the hypothetical role of the serotoninergic system in the genesis of schizophrenia, special attention has been paid to the drugs selectively acting on this system. The results of the use of fluoxetine as co-treatment with neuroleptics, published by various authors have been presented. PMID- 8722242 TI - [Cost analysis of treating depression. Comparison of tricyclic antidepressants and selective serotonine re-uptake inhibitors]. AB - The question of costs in treating the most widely spread diseases has appeared in medical literature for several years. A task of modern health economics is to estimate and, if possible, to reduce direct and indirect costs of illnesses having social dimension. Since depression takes important place among them, it also stays in a circle of economists' interests. Latest changes in pharmacotherapy of depression, connected with appearence, by tricyclics, selective serotonine reuptake inhibitors, seem to be very promising: they have similar antidepressive effectiveness, but differ in toxicity after overdose, quantity and quality of side-effects, treatment dropout. However the organisation of Polish health service still passes over this economic aspect, it is probable that this situation will change in the nearest future, so it can be interesting to approach the methods of economic thinking in western psychiatric services. PMID- 8722243 TI - [Pharmacotherapy of depression in patients with and without conversion from depression to mania in the course of bipolar affective disorder]. AB - To answer the question whether the kind and duration of the applied antidepressant therapy is a risk factor for the occurrence of mania directly after depression, the treatment of depression relapses in 30 patients with switch from depression to mania in the course of bipolar disease, and the treatment of 30 (appropriately selected) patients without this change was compared. No differences in treatment procedures were stated in comparable relapses, apart from more frequent application of lithium carbonate and carbamazepine in the patients with the switch from depression to mania. It was shown that long-term application of antidepressant drugs is not connected with the increased risk of such change. PMID- 8722244 TI - [Pharmacotherapy in depression during conversion from depression to mania in patients with bipolar affective disorder]. AB - Frequency of occurrence of mania directly after depression during antidepressant treatment with drugs of different pharmacological characteristics was examined. 210 depressive episodes (in 118 patients) finished with switching from depression to mania were analyzed. It was found that this phenomenon occurs more frequently during therapy with classical tricyclic drugs (amitryptyline, imipramine) than with the second generation antidepressant drugs. A group of patients in whom the switch from depression to mania occurs independently of the kind of applied therapy, was observed. PMID- 8722245 TI - [Mianserin efficacy in the treatment of depression]. AB - Fifty patients suffering from depression were treated wigh mianserin in monotherapy. ICD-9 and DSM-III criteria for depression were used. Patients were divided into four groups--with monopolar depression (28 patients), bipolar depression (8 patients), organic depression (10 patients), neurotic depression (4 patients). The intensity of psychopathological symptoms of depression was established using the Hamilton Depression Rating Scale (HDS) on the 7th, 14th and 28th day of the treatment. The antidepressant action of mainserin was evident already on the 14th day of treatment. Mianserin proved to be most effective in endogenous bipolar depression group and neurotic depression group (70% reduction in the score obtained on the HDRS). Mianserin was well tolerated by most patients. Most frequent side effects observed were: hypertension (8 patients), feeling of anxiety (10 patients), constipation (8 patients), tachycardia (6 patients), dry mouth (3 patients). PMID- 8722246 TI - [Therapeutic problems in the application of antiparkinsonian drugs in psychiatry]. AB - This paper describes therapeutic problems concerning antiparkinsonian drugs in psychiatry. Particular attention was paid to problems like: 1. Possibility of making a therapeutic mistake of too prolonged and therefore harmful pharmacotherapy with antiparkinsonian drugs in the treatment of most common psychoses. 2. The importance of a careful selection of an optimal antiparkinsonian drug, especially in psyhogeriatric cases (for example Amantadinum). 3. The description of other indications for antiparkinsonian drugs: affective diseases or alcoholic withdrawal syndroms. PMID- 8722247 TI - [Can psychiatric examination be useful in early diagnosis of intracranial expansion processes?]. AB - The authors carried out pilot investigations of the role of basic psychiatric examination, paying special attention to psychiatric evaluation of anamnestic data in the early recognition of intracranial expansion processes (intracranial tumors). The investigations show that psychiatric diagnostic is not sufficiently effective in the case of intracranial tumors. Despite rich symptomatology of the advanced disease processes, the early symptoms are ambiguous and "litle alarming". What occurs earlier is the "strong" neurological symptoms that can be verified with the use of more and more accessible techniques (mainly neuropicturing). PMID- 8722248 TI - [Serum cholesterol, triglycerides, HDL and LDL in aggressive elderly patients with dementia]. AB - Associations were analysed between serum concentrations of cholesterol, triglycerides, HDL and LDL measured after hospital admission, and physical aggression in a sample of elderly patients with dementia (210 women and 160 men). A significant lower serum cholesterol and LDL concentration were found in aggressive patients of both sexes and a significant lower serum triglycerides only in aggressive women. In the subgroups of Alzheimer type dementia, women showed significant lower serum triglycerides, and aggressive men significant lower LDL concentration compared with non-aggressive controls. There were no significant differences in serum HDL between aggressive and non-aggressive patients. PMID- 8722249 TI - [Comments from the conference "Understanding social phobia". Oslo, September 15 16, 1995]. PMID- 8722250 TI - [International conference of mentally disordered workers. Liege (Belgium). August 28-September 8, 1995]. PMID- 8722251 TI - [Non-governmental organizations for the social assistance to people with mental disorders]. PMID- 8722253 TI - Fluoride exposure in cervids inhabiting areas adjacent to aluminum smelters in Norway. I. Residue levels. AB - Mandibular fluorine concentrations were determined in 1,425 red deer (Cervus elaphus), 240 moose (Alces alces), and 424 roe deer (Capreolus capreolus) collected in Norway from 1990 to 1993 in seven municipalities in which aluminum smelters are located, in eight neighboring municipalities, and in eight reference areas representing background levels. Background fluorine concentration was significantly correlated with age in all three species. Roe deer had the highest mean background fluorine level in each age group, followed by red deer. Due to differences in fluoride exposure, large variations in bone fluorine residues were evident between locations. In Ardal, the district most severely exposed to fluoride contamination, nine of ten cervids had fluorine concentrations exceeding background levels. The proportions of red deer with fluorine residues exceeding background levels also were high in neighboring municipalities to Ardal. We propose that roe deer are a better biomonitor of local fluoride exposure than red deer and moose, due to their more sedentary behavior. PMID- 8722254 TI - Fluoride exposure in cervids inhabiting areas adjacent to aluminum smelters in Norway. II. Fluorosis. AB - Mandibles from 1104 red deer (Cervus elaphus), 147 moose (Alces alces), and 453 roe deer (Capreolus capreolus), collected between 1990 and 1993 in the vicinity of seven Norwegian aluminum smelters, were examined for dental fluorotic and osteofluorotic lesions. The metacarpal or metatarsal bones from 214 of these cervids also were evaluated. Dental fluorotic lesions occurred in all three cervid species. Prevalence of dental fluorosis was generally low at the various locations, with the exception of Ardal, where 15% of the cervids examined were affected. Only sporadic cases of severe dental fluorotic lesions were diagnosed. All red deer yearlings (1.5 yr) with mandibular fluorine (F) levels exceeding 2,000 ppm F, had dental fluorosis. However, the lowest skeletal fluorine level found in a fluorotic animal of this age was 1,355 ppm F. Gross osteofluorosis occurred in only three cervids, all with mandibular fluorine residues > 8,000 ppm F. Hence, generalized fluorosis was not a prominent feature in the material studied. PMID- 8722255 TI - Fluoride exposure and selected characteristics of eggs and bones of the herring gull (Larus argentatus) and the common gull (Larus canus). AB - Fluorine concentrations were determined in the shell of 285 herring gull eggs (Larus argentatus) and 120 common gull eggs (Larus canus), collected May 1991 to 1993, from breeding colonies exposed to emissions from two Norwegian primary aluminum smelters located at Karmoy and Sunndal, and from unexposed reference localities in Eigersund, Sola, and Stavanger. Volume-index, shell thickness, thickness-index, and fertilization of the eggs also were monitored. In both species, the shell fluorine concentration was significantly increased in eggs collected at sites exposed to fluoride emissions. No effects on other egg characteristics were observed. In both exposed and unexposed sites, the last-laid egg in a clutch, normally containing three eggs, had the highest shell fluorine residue. Fluorine levels also were analyzed in femurs from 42 herring gulls, collected from Karmoy and Sola in May 1993. The relationship between sex and fluoride accumulation, and the relations between fluorine concentration in femurs of laying herring gulls and in the shell of their eggs, were evaluated. Bone morphology also was studied. Bone fluorine concentrations were raised significantly in emission-exposed female birds. Moreover, females from the exposed site had significantly higher fluorine residues than males. There was a positive correlation between fluorine levels in femurs of individual laying birds and those in the shells of their eggs. No changes in bone morphology due to fluoride exposure was found. PMID- 8722256 TI - Immunotoxicity studies in mink (Mustela vison) chronically exposed to dietary bleached kraft pulp mill effluent. AB - The immunotoxic potential of bleached kraft pulp mill effluent (BKME) to cell mediated immunity in mink (Mustela vison) was investigated October 1993 through May 1994. For 26 weeks, 20 mink were fed a diet based upon fish caught within 6 km downstream of a bleached kraft mill in Saskatchewan, Canada. Water for this group contained 25% softwood-run BKME. Twenty control mink were fed nutritionally matched diets based upon fish from lakes receiving no municipal or industrial effluent and tap water. Using in vitro and in vivo immunotoxicity assays, the proliferative response of mink peripheral blood mononuclear cells (PBMC) to mitogens was optimal, at 72 hr with 10 micrograms/ml Concanavalin A, 1/80 dilution pokeweed mitogen, and 1/80 dilution phytohemagglutinin. Bacterial cell wall Escherichia coli lipopolysaccharide did not stimulate mitosis of the mink PBMC. No difference (P < 0.05) in PBMC proliferation was seen between the control and BKME-exposed mink with any of the mitogens used. Delayed type hypersensitivity (DTH), a cell mediated response, was assessed in mink vaccinated with live bacille Calmette-Guerin (BCG) and then challenged by intradermal toe web injection with 200 micrograms of sonicated BCG approximately 6 weeks later. The DTH response in the BKME-exposed mink was impaired based upon assessment using skin thickness measurements, histopathological assessment and image analyzer technology. This decreased response is evidence for suboptimal immune function associated with BKME exposure, which could affect the competitive fitness of piscivorous mammals naturally exposed to BKME. PMID- 8722257 TI - Effects of Prudhoe Bay crude oil on hatching success and associated changes in pipping muscles in embryos of domestic chickens (Gallus gallus). AB - Fertile white leghorn chicken eggs were exposed to 0, 1, 2, 4, 6, 8 or 16 microliters of Prudhoe Bay Crude oil (PBCO) on day 9 of incubation. The effects of oil on pipping and hatching success, body weight gain after hatching, serum creatine kinase levels, and pathological changes in organ systems were assessed in embryos that had survived acute toxic effects and were alive on day 18 of incubation. Exposure to oil greatly reduced pipping and hatching success. Severe edema and hemorrhage in the pipping muscle, multifocal subcapsular hepatic necrosis, marked depletion of lymphocytes in the bursa of Fabricius with infiltration by heterophils, and occasional dorso-caudal subcutaneous edema were observed in treated embryos. Pipping muscles were heavier in oil-exposed embryos. Embryos exposed to 4 microliters of PBCO had significantly reduced gain in body weight post-hatching. Serum creatine kinase levels were significantly elevated in the oil-exposed embryos only at the time of hatching. There was no evidence that exposure to oil caused degenerative changes in pipping muscle cells. PMID- 8722258 TI - Further studies of brain cholinesterase: cholinergic receptor ratios in the diagnosis of acute lethal poisoning of birds by anticholinesterase pesticides. AB - Cholinesterase (ChE) and the muscarinic (mAChR) and nicotinic (nAChR) cholinergic receptors were measured in the brains of 86 birds of 20 different species collected in Saskatchewan, Canada during 1991 and 1992. There was a strong correlation between ChE and mAChR, and the ratio of ChE: mAChR was 38% less variable than was ChE alone. In a hypothetical test for minimal acute lethal poisoning of all birds based on a reduction by 50% of the normal ChE measured in each, the use of a single diagnostic threshold value of the ChE: mAChR ratio had a diagnostic sensitivity of 98% and a specificity of 99%, thus making theoretically possible the diagnosis of acute lethal poisoning by anticholinesterase chemicals without the use of reference control values for ChE. Both ChE and mAChR were highly stable during 12 days of postmortem decomposition under several different daily temperature regimes. Substantial changes in these molecules occurred only under conditions of constant high temperature (36 C). Acute lethal poisoning by the organophosphate chlorpyrifos did not affect the density of mAChR. Postmortem decomposition did not appear to be an important confounding factor in the diagnostic interpretation of either ChE or the ChE: mAChR ratio except under hot climatic conditions. PMID- 8722259 TI - Prouterina wescotti n. gen., n. sp. (Trematoda: Prouterinidae N. Fam.) from the brain, lungs, and nasal sinuses of a black bear (Ursus americanus) from Idaho. AB - Prouterina wescotti gen. n. and sp. n. (Trematoda: Prouterinidae N. Fam.) is described from a free-ranging black bear (Ursus americanus) which died in May 1995 in northern Idaho (USA). Adult digenetic trematodes were detected in brain, lungs, and nasal sinuses, and were likely responsible for the emaciated condition, copious nasal discharge, neurological signs, and death of the bear. Mature trematodes recovered from the bear were conical with small spines on the tegument. The anterior end was broad and tapered gradually toward the posterior. Mean (+/-SE) size of the mature trematodes was 3.67 (+/-0.08) by 2.14 (+/-0.04) mm (n = 80). Eggs are operculated, gold, and 68.2 (+/-0.42) by 41.4 (+/-0.41) microns (n = 75). Suckers are well developed and located in the anterior half of the body, with the genital pore just posterior to the ventral sucker. Testes are tandem and the ovary is lateral and slightly anterior to the anterior testes just posterior to the ventral sucker. The uterus is predominantly anterior to the ventral sucker and is the most distinctive feature of the trematode. PMID- 8722260 TI - Efficacy and safety of naltrexone hydrochloride for antagonizing carfentanil citrate immobilization in captive Rocky Mountain elk (Cervus elaphus nelsoni). AB - We evaluated efficacy and safety of naltrexone for antagonizing carfentanil immobilization in 12 captive Rocky Mountain elk (Cervus elaphus nelsoni) using a randomized incomplete block experiment. In three replicate trials, elk were hand injected with 10 micrograms carfentanil citrate/kg body weight intramuscularly. Fifteen min after each elk became recumbent, we administered naltrexone HCl (25% of dose intravenously, 75% subcutaneously) dosed at 0 (control), 25, 50, or 100 mg/mg carfentanil; after an additional 15 min of immobilization, controls received 500 mg naltrexone HCl/mg carfentanil. Elk were immobilized in 34 of 36 attempts; the mean (+/-SE) induction time was 3.1 +/- 0.2 min. Regardless of dose, all elk stood < 9 min after receiving naltrexone; controls remained immobilized until they received antagonist. Mean recovery times did not differ with increasing naltrexone dose (P = 0.31) or among individuals (P = 0.16). None of the elk receiving 100 or 500 mg naltrexone/mg carfentanil renarcotized, but three of eight and seven of nine elk receiving 50 and 25 mg naltrexone/mg carfentanil, respectively, showed signs of mild renarcotization 8 to 24 hr later (P = 0.0002). We observed no adverse clinical effects in elk receiving < or = 500 mg naltrexone/mg carfentanil. Based on these data, we recommend 100 mg/mg carfentanil as a minimum effective dose for rapidly antagonizing immobilization and preventing renarcotization. PMID- 8722261 TI - Forensic (medico-legal) necropsy of wildlife. AB - Post-mortem examination of wild animals for legal purposes has become increasingly common. Special procedures are necessary during such necropsies to ensure that the information collected is suitable for use in a court of law. Forensic necropsies should be performed by pathologists with formal training and experience, because these credentials will be examined if a case reaches court. There must be strict attention to maintaining a chain of custody when specimens are received from the field and in all subsequent procedures, so that the identity of specimens and information resulting from the necropsy is beyond question. A complete record must be kept of all procedures and observations. The necropsy report must be detailed, clear and, as far as possible, written in non technical language. Photography should be used to preserve visual evidence. Special precautions are required for collecting specimens, such as bullets, suspected toxins, and material for DNA analysis, and for sending these specimens to other laboratories. PMID- 8722262 TI - Metastatic carcinoma of probable transitional cell origin in 66 free-living California sea lions (Zalophus californianus), 1979 to 1994. AB - Sixty-six (18%) cases of widely metastatic carcinoma of probable transitional cell origin were identified in 370 California sea lions (Zalophus californianus) stranded alive along the central California (USA) coast, between January 1979 and December 1994. Live animals were usually emaciated and anorectic, with perineal edema and occasionally hind-flipper paralysis or paresis. Large yellow caseous masses were observed in the sub-lumbar lymph nodes, often extending around the ureters resulting in hydroureter. Histologically, metastases were usually widespread, and the primary neoplastic focus undetectable. This is the highest reported prevalence among necropsied animals of neoplasia in a pinniped population to date. PMID- 8722263 TI - Prevalence and histopathology of shell disease in turtles from Lake Blackshear, Georgia. AB - Turtles in Lake Blackshear, Crisp County, Georgia (USA) were evaluated for shell disease during intensive trapping efforts on 8 and 9 May 1990. The disease was most prevalent in Pseudemys concinna (74%) and Trachemys scripta (35%). The degree of necrosis on the carapace was significantly positively correlated with the degree of necrosis on the plastron in T. scripta (rs = 0.50), but not in P. concinna (rs = 0.06). Female T. scripta with lesions were significantly larger than females without lesions. Lesions were not detected on six other species of turtles. Some areas contained multinucleate osteoclasts that were destroying bone. No tumors were detected in soft tissue samples. PMID- 8722264 TI - Causes of owl mortality in Hawaii, 1992 to 1994. AB - Eighty-one barn owls (Tyto alba) and five Hawaiian owls or pueo (Asio flammeus sandwichensis) from Kauai, Oahu, Lanai, Molokai, Maui and Hawaii (USA) were evaluated for cause of death, November 1992 through August 1994. The most common cause of death in barn owls was trauma (50%) followed by infectious disease (28%) and emaciation (22%). Most traumas apparently resulted from vehicular collisions. Trichomoniasis was the predominant infectious disease and appeared to be a significant cause of death in barn owls in Hawaii. Pasteurellosis and aspergillosis were encountered less commonly. No predisposing cause of emaciation was detected. Stomach contents from 28 barn owls contained mainly insects (64%) of the family Tetigoniidae and Gryllidae, and rodents (18%); the remainder had mixtures of rodents and insects or grass. Three pueo died from trauma and one each died from emaciation and pasteurellosis. We found no evidence of organochlorine, organophosphorus, or carbamate pesticides as causes of death in pueo or barn owls. PMID- 8722265 TI - Serological survey of small mammals in a vesicular stomatitis virus enzootic area. AB - Small mammals were captured in a Costa Rican dairy farm located in a vesicular stomatitis virus (VSV) enzootic focus, in order to determine which species were naturally infected by this virus. Monthly captures were performed from March 1989 to February 1990. Eighty-four individuals belonging to the orders Rodentia (n = 52), Insectivora (n = 31) and Marsupialia (n = 1) were captured. Only Sigmodon hispidus had neutralizing antibodies to VSV; among 21 animals, six had antibodies to Indiana, one to New Jersey, and two to both serotypes. In addition, groups of 40 sentinel mice (Mus musculus, strain C3H) were placed in cages distributed throughout the farm. Each group was exposed for 1 mo over a period of 1 yr. None of 312 sentinel mice developed antibodies against either VSV serotype. Based on these results, we believe that S. hispidus might be part of the natural cycle of VSV in this enzootic focus. Caged Mus musculus do not seem appropriate for monitoring VSV activity in this area. PMID- 8722266 TI - European brown hare syndrome in free-ranging hares in Poland. AB - A study of European brown hare syndrome (EBHS) was conducted in Poland (Czempin). From April 1993 until February 1994, 100 blood and 78 spleen samples of European brown hares (Lepus europaeus) were tested for prevalence of EBHS and rabbit hemorrhagic disease (RHDV) antibodies and EBHS virus antigen with two enzyme linked immunosorbent assay (ELISA) test kits. Thirty-eight of 100 serum samples were positive for EBHS, and six (7.6%) of 78 of the spleen materials were-antigen positive for EBHS virus. Three (3%) of the sera were positive against RHDV, whereas two of these also were seropositive for EBHSV. European brown hare syndrome virus seropositive hares were most frequently found from April to September. Based on negative staining electron microscopy investigations of liver and spleen homogenates of all antigen-positive hares, we observed caliciviruses in only one animal. For histopathological investigations organ specimens were available from 98 hares. Histopathological findings corresponded with the clinical picture of chronic EBHS. A pathohistological picture consistent with EBHS was found in 22 (22%) of investigated hares and corresponded in 50% of the animals which reacted positively in the EBHSV antigen-ELISA and in 29% of the animals which reacted positively in the EBHSV antibody-ELISA. These results from western Poland are the first that caliciviruses are present in European brown hare population in Eastern Europe and may be one of the causes for increased mortality in the Polish hare population over the past 10 years. PMID- 8722267 TI - Effects of visible signs of contagious ecthyma on mass and survival of bighorn lambs. AB - External signs of contagious ecthyma became common in a population of Rocky Mountain bighorn sheep (Ovis canadensis) in Alberta, Canada, after it attained high density. Between 1990 and 1993, we studied effects of this disease on mass gain and survival of lambs. Prevalence and severity were independent of lamb sex. Lambs with large sores and scabs gained less mass than other lambs and were lighter the following spring as yearlings. There was no significant effect of the disease upon lamb survival, and contagious ecthyma did not appear to play a primary role on the dynamics of the study population. PMID- 8722268 TI - Susceptibility of selected rodent species from Colorado to Borrelia burgdorferi. AB - To determine the susceptibility of some common Colorado (USA) rodent species to Borrelia burgdorferi, pregnant Peromyscus maniculatus, Tamias minimus, and Spermophilus lateralis were trapped in May 1990 and kept in quarantine until their young were old enough to be used in the experiment. Six to eight 8-wk-old individuals of each of the Colorado species and, for comparison, eight laboratory raised P. leucopus were subcutaneously inoculated with > or = 10(5) spirochetes in 0.1 ml in July 1990. Tissue specimens were collected for isolation from these animals through April 1991. Spirochetes were isolated from blood, ear, bladder, kidney, spleen, liver, and eye in Barbour-Stoener-Kelly (BSK) medium from P. maniculatus, P. leucopus and T. minimus. Spirochetes were isolated from at least one tissue from all of these animals and no isolations were obtained from any of the S. lateralis. Thus, three of the four rodent species tested are susceptible to, and could harbor, B. burgdorferi. PMID- 8722269 TI - Experimental infection of the raccoon (Procyon lotor) with Borrelia burgdorferi. AB - The reservoir competence of the raccoon (Procyon lotor) for the Lyme disease spirochete (Borrelia burgdorferi) was evaluated in the laboratory during September 1991 to April 1993. Five raccoons were exposed to spirochete-infected (JD1 and Wisconsin 210 Wise strains) Ixodes scapularis nymphs (20/raccoon). A second feeding of spirochete-infected (Wisconsin 210 Wise strain) nymphs (20/raccoon) was performed with four of the original raccoons. Xenodiagnosis with cohorts of I. scapularis larvae (300/cohort) or nymphs (150/cohort) that were periodically placed on each animal was used to detect infection. We examined 1943 engorged ticks by a indirect immunofluorescence monoclonal antibody assay, but no spirochetes were detected. After exposure to spirochete-infected ticks, blood samples were collected at approximately weekly intervals and ear-skin biopsy samples were taken from each animal every third week. These tissues were placed in Barbour-Stoenner-Kelly media. Spirochetes were isolated in cultures of skin (wk 3, 5, 9, 81, and 83) and blood (wk 5, 8, 9, 11, and 12) of one raccoon and the skin (wk 28 and 31) of another raccoon. Antibody response of each animal was monitored through enzyme-linked immunosorbent assays and immunoblotting of blood serum against B. burgdorferi proteins. Except for one animal, raccoons did not have an antibody response until they were fed upon by a second cohort of infected I. scapularis nymphs. Based on Western blot analyses, raccoons exposed to B. burgdorferi via tick bite responded to the 31- (OspA) and 34-KDa (OspB) antigens. Response to other antigens varied among raccoons. Based on our results raccoons may be inefficient reservoirs for B. burgdorferi. Although some raccoons can become infected with B. burgdorferi, they may not transfer the infection to attached ticks. PMID- 8722270 TI - Attempted protection of bighorn sheep (Ovis canadensis) from pneumonia using a nonlethal cytotoxic strain of Pasteurella haemolytica, biotype A, serotype. AB - Between February and April, 1994, we tested the hypothesis that bighorn sheep (Ovis canadensis canadensis) inoculated with a cytotoxic isolate of Pasteurella haemolytica biotype A, serotype 11 (A11) could withstand challenge inoculation with a cytotoxic strain of P. haemolytica A2 of domestic sheep origin known to cause lethal pneumonia in bighorn sheep. On experimental day O, two bighorn sheep were inoculated intratracheally with 6 x 10(9) colony forming units (cfu) of a cytotoxic strain of P. haemolytica A11 (group 1); two bighorn sheep were inoculated intratracheally with 6 x 10(9) cfu of a noncytotoxic P. haemolytica A11 (group 2), and two control bighorn sheep were inoculated intratracheally with a similar volume of brain heart infusion (BHI) broth (group 3). After inoculation, all bighorn sheep remained healthy. On experimental day 16, group 1 bighorn sheep each were given the same intratracheal inoculation as on day O, and groups 2 and 3 bighorn sheep each were inoculated with BHI broth at the same volume as group 1. All bighorn sheep remained healthy following inoculations. On experimental day 42, bighorn sheep in groups 1 and 3 each were challenged with an intratracheal inoculation of 6 x 10(9) cfu of P. haemolytica A2 of domestic sheep origin known to be lethal in bighorn sheep. Group 2 sheep each were inoculated intratracheally with BHI broth at the same volume as groups 1 and 3. The four bighorn sheep in groups 1 and 3 that received the challenge inoculation died from acute bronchopneumonia within 72 hours after challenge inoculation, and cytotoxic P. haemolytica A2 was isolated from the four dead bighorn sheep. Both cytotoxic or noncytotoxic strains of P. haemolytica A11 were not lethal and did not cause pneumonia in the experimentally inoculated bighorn sheep. However, previous inoculation with cytotoxic P. haemolytica A11 did not protect the bighorn sheep against later experimental challenge inoculation with a known lethal strain of cytotoxic P. haemolytica A2 under the conditions defined in these experiments. PMID- 8722271 TI - Isolation of Pasteurella spp. from free-ranging American bison (Bison bison). AB - From November 1991 through March 1992, nasal and pharyngeal swab samples were collected from 45 bison (Bison bison) from Yellowstone National Park, Montana (USA) and cultured for Pasteurella spp. Thirteen isolates of Pasteurella spp. were recovered from 10 (22%) of the animals. Ten isolates were from pharyngeal samples in contrast to three isolates from nasal samples. Pasteurella haemolytica (six biotype T, two biotype A, and two biotype 3) was the predominant Pasteurella species. Five biotype T isolates were serotype 4 and the sixth agglutinated in antisera 3, 4, and 10. Both biotype A isolates were untypable with antisera to recognized type strains. Pasteurella multocida was isolated from the pharyngeal samples of one animal. Two isolates could not be identified to species. PMID- 8722272 TI - Infectious keratoconjunctivitis in free-ranging mule deer (Odocoileus hemionus) from Zion National Park, Utah. AB - An epizootic of infectious keratoconjunctivitis (IK) was studied opportunistically in free-ranging mule deer (Odocoileus hemionus) from Zion National Park, Utah (USA), from November 1992 to March 1994. Moraxella sp. and Chlamydia sp. were isolated from the conjunctiva of two of seven deer. In addition, Thelazia californiensis occurred on the conjunctivas of six of seven deer. Based on field observations, adults appeared to be affected clinically at a higher incidence during both years as opposed to juveniles. Corneal opacity was the most apparent clinical sign from 1992 to 1993. However, in the following year, blepharospasm and epiphora were noted more often. We were also able to document the clinical recovery of three affected deer. In addition, Moraxella sp. was recovered from the eyes of a clinically unaffected deer 1 year after the epizootic occurred. PMID- 8722273 TI - Exposure of wild waterfowl to Mycoplasma anatis. AB - We developed an ELISA procedure to assess the presence of M. anatis-specific serum antibody in ducks. Sera from exposed and unexposed Pekin ducks (Anas platyrhynchos) were used to standardize the ELISA and to establish reference ranges to classify ELISA results as exposed or not exposed. We conducted serological surveys of female waterfowl in the central and eastern United States between 1988 and 1992 to assess the frequency of exposure in wild waterfowl. Adult breeding mallards (Anas platyrhynchos), wintering mallards, and black ducks (Anas rubripes) had high prevalences of exposure to M. anatis (25% to > 80%). In comparison, none of the breeding adult canvasbacks (Aythya valisineria) had serum antibody levels indicating exposure. Approximately 50% of the juvenile mallards and black ducks were exposed to M. anatis by 8 months of age, indicating high transmission rates among wild birds. PMID- 8722274 TI - First record of a louse fly, Stilbometopa impressa (Bigot), and new host for Microlynchia pusilla (Speiser) (Hippoboscidae) from the Cape Region, Baja California Sur, Mexico. AB - Nine of thirty California quail (Callipepla californica achrustera) captured in autumn of 1992, 17 km west of La Paz, Baja California Sur, Mexico, were parasitized by louse flies. We identified eight Microlynchia pusilla and three Stilbometopa impressa from 30 quails in the ratio of 2.75:1. These are the first records of S. impressa for Cape Region and the first time either fly has been reported from the California quail in Baja California Sur. PMID- 8722275 TI - Intrahepatic larval nematode infection in the northern spring peeper, Pseudacris crucifer crucifer (Anura: Hylidae), in West Virginia. AB - Larval stages of an unknown nematode were observed encapsulated in the livers of spring peepers, Pseudacris crucifer crucifer (Weid-Neuweid), collected from a marsh in western West Virginia (USA) during the spring breeding seasons of 1993 and 1994. Prevalence and mean intensity of infection were 37% (30 of 82 animals) and 2.03 parasites per infected host, respectively. Capsules with white or darkly pigmented walls were observed in infected livers; the former containing viable larvae, and the latter enveloping larvae in various stages of degeneration. PMID- 8722276 TI - Protozoan parasites and pathological findings in Hydromys chrysogaster (Muridae: Hydromyinae) from Queensland. AB - Tissues from 23 Australian water rats (Hydromys chrysogaster) collected from five localities in central and northern Queensland, Australia, between February 1992 and May 1993, were examined for protozoan parasites and additional pathological changes. We found Klossiella hydromyos in the kidneys, Toxoplasma gondii in the brain and skeletal muscles and Sarcocystis sp. in the somatic musculature. Other pathological findings, including interstitial nephritis, interstitial pneumonia and a tongue abscess, as well as helminth-induced lesions in the lungs, mesenteries, stomach wall and cecal wall were also noted. PMID- 8722277 TI - Seroprevalence of orthopox virus specific antibodies in red foxes (Vulpes vulpes) in the Federal State Brandenburg, Germany. AB - The prevalence of orthopox virus (OPV)-specific antibodies in 1,040 red foxes (Vulpes vulpes) was evaluated on a large scale in the German Federal State Brandenburg. Serum samples were selected from 809 communities within the study area from January 1991 to September 1994 by simple random sampling. Screening was carried out by an indirect enzyme-linked immunosorbent assay (ELISA). Orthopox virus-specific antibodies were found in 162 (16%) of the 1,040 fox sera. Furthermore 154 (15%) sera were considered suspect positive. The specificity of the antibodies detected in ELISA-positive and suspect positive sera was confirmed by Western blotting. Presence of OPV-antibodies occurred in 291 communities. No correlation of OPV-antibodies findings to latitude or characteristic topographical and ecological peculiarities of the study area was found. Although the causative agent is still unknown we believe that orthopox viruses probably have a ubiquitous presence among red foxes. PMID- 8722278 TI - Serological evidence for zoonotic hantaviruses in North Carolina rodents. AB - In a survey of seven species of wild rodents (n = 423) collected between October 1993 and March 1994 from the three principal ecological biomes of North Carolina (USA), we found hantavirus antibodies in seven (2%) of 301 Peromyscus spp. Hantavirus antibodies were detected in P. leucopus and P. maniculatus captured from mountain and coastal island biomes. Three mice were positive for Sin Nombre virus, while four others had antibodies to Seoul virus or a related agent. Two mice serologically positive for Sin Nombre virus were collected from inside a private mountain domicile. We conclude that the risk of human exposure to hantaviruses in North Carolina resembles that for most other areas of the continental United States. PMID- 8722279 TI - Assessment of humoral immune response in mink (Mustela vison): antibody production and detection. AB - A method for investigating the humoral immune response in mink (Mustela vison) was developed between October 1993 and March 1994. Protein A, 1:8000 dilution, had a high affinity for mink immunoglobulin, while anti-ferret (Mustela putorius) antibody, 1:200 dilution, had a weaker affinity. Four adult mink were immunized with a hapten, dinitrophenol (DNP), conjugated to a large carrier protein, keyhole limpet hemocyanin (KLH), and received two boosters at 3-week intervals. This provoked a strong T-lymphocyte dependent humoral immune response. An indirect enzyme linked immunosorbent assay (ELISA) was used to quantify the antibody produced. All mink had undetectable anti-DNP-KLH antibody in the pre immune sera, with antibody levels increasing post-immunization, and peaking after the first or second booster. PMID- 8722280 TI - Isolation of Yersinia pseudotuberculosis and Listeria monocytogenes serotype 4 from a gray fox (Urocyon cinereoargenteus) with canine distemper. AB - In February 1993, clinical, pathological, and microbiological investigations were performed on an adult female gray fox (Urocyon cinereoargenteus) from northern Mississippi (USA). The fox had clinical signs consistent with canine distemper virus encephalitis. Eosinophilic inclusions characteristic of canine distemper virus were in the nuclei and cytoplasm of cerebral neurons and glial cells and in the cytoplasm of urinary, gastric, pancreatic and biliary epithelial cells. The liver contained multifocal microscopic nodular foci of granulomatous to pyogranulomatous inflammation and necrosis with large colonies of small Gram negative coccobacilli. A low number of small Gram-positive bacilli were within viable-appearing Kupffer cells and hepatocytes. Yersinia pseudotuberculosis and Listeria monocytogenes serotype 4 were isolated from the liver and a mesenteric lymph node. PMID- 8722281 TI - Myocardial Purkinje degeneration and necrosis with fibrosis in free-ranging black rhinoceroses (Diceros bicornis) in Zimbabwe. AB - Degeneration and necrosis of Purkinje fibers with fibrosis around Purkinje fibers were found in the hearts of three adult black rhinoceroses (Diceros bicornis) in Zimbabwe in 1989 and 1990, among 38 animals examined from 1988 to 1994. Causes of death were not apparently related to these changes, nor was there evidence of heart failure. The etiology of these changes is unknown. PMID- 8722282 TI - Synovial chondromatosis in a great horned owl (Bubo virginianus). AB - A case of synovial chondromatosis in a great horned owl (Bubo virginianus) was found in June 1993. In radiographs of bilateral swelling of the scapulohumeral joint we observed numerous mineralized foci in the soft tissue. The foci were identified by light microscopy as cartilaginous metaplasia. This is the first report of synovial chondromatosis in an owl. PMID- 8722283 TI - Pyogranulomatous peritonitis associated with Nocardia sp.-like organisms in a raccoon (Procyon lotor). AB - During 1992 raccoons (Procyon lotor) were live-trapped in northeastern Pennsylvania (USA). In one of these animals a localized pyogranulomatous peritonitis was seen. Grossly a large mass with a central area of liquifactive necrosis was present in the anterior abdomen. Microscopically the lesion contained multiple colonies of filamentous organisms with histomorphologic and histochemical charcteristics resembling Nocardia sp. This appears to be the first report of Nocardia infection in the raccoon. PMID- 8722284 TI - Dental and mandibular injury in an Antarctic fur seal, Arctocephalus gazella, at Heard Island, Southern Ocean. AB - The skull of an adult male Antarctic fur seal (Arctocephalus gazella) collected at Heard Island, Southern Ocean, October 1992, had chronic changes attributable to a fracture of the left lower canine, luxation of the mental symphysis, osteomyelitis of the left and right mandibles, and periostitis of the left maxilla. PMID- 8722285 TI - Hemolytic anemia in wild seaducks caused by marine oil pollution. AB - Clinico-pathological examinations were conducted on wild white-winged scoters (Melanitta fusca) contaminated with fuel oil (Bunker C oil) from a capsized cargo ship in February 1993 in Japan. The erythrocyte count, hemoglobin concentration and hematocrit value in the oiled seaducks all were decreased and numerous immature erythrocytes were observed in blood smears. In addition, hemosiderosis was observed in the liver, kidney, and lung of some birds. We propose that the sea-ducks suffered from hemolytic anemia induced by ingestion of oil, which occurs when the birds preen their oiled plumage. PMID- 8722286 TI - Winter poisoning of coyotes and raptors with Furadan-laced carcass baits. AB - Three bald eagles (Haliaeetus leucocephalus), a red-tailed hawk (Buteo jamaicensis), and two coyotes (Canis latrans) found in a field in north-central Kansas (USA) in December 1992 were poisoned by flowable carbofuran (Furadan 4F) placed on sheep (Ovis aries) carcasses to kill coyotes. The carbofuran was placed on the carcasses in October 1992, but the coyotes and raptors apparently were killed in late December. Thus, flowable Furadan can cause direct and secondary deaths of wildlife under some circumstances for at least 60 days following placement. PMID- 8722287 TI - Environmental, age, and sex effects on cotton rat (Sigmodon hispidus) hematology. AB - We determined the effects of sex, age, and environment (inbred, captive-wild, and wild animals) on selected blood parameters of the cotton rat (Sigmodon hispidus) in central Oklahoma (USA) from 1990 to 1994. Male and female cotton rats had similar blood profiles. Age-related differences were confined to differential white blood cell counts where adults possessed greater numbers of neutrophils and lower numbers of lymphocytes compared to juveniles. Environment had a strong influence on many hematological parameters. Hematocrit, hemoglobin concentration, RBC count, and eosinophil number were generally greater for wild stocks compared to inbred animals, and differences were more pronounced for adults than juveniles. PMID- 8722288 TI - The attractiveness of a liquid bait to white-tailed deer in the Central Appalachian Mountains, Virginia, USA. AB - We determined the attractiveness of a bait composed of apple juice, apple odor, water, glycerin, and salt to captive white-tailed deer (Odocoileus virginianus) at the National Zoological Park Conservation and Research Center, Front Royal, Virginia, USA. Thirty-nine deer (three adult males, 20 adult females, eight male fawns, eight female fawns) were presented with the bait formulation in a 22-ha enclosure during January, February, and May 1994. Daily consumption, and the frequency of visits to bait dispensers and the frequency of drinking bouts were recorded. Overall consumption was greatest in May (P < 0.001). While adults and fawns visited bait dispensers equally often, fawns drank more frequently (P < 0.001). PMID- 8722289 TI - Antagonism of xylazine in white-tailed deer with intramuscular injection of yohimbine. AB - Eighteen free-ranging white-tailed deer (Odocoileus virginianus) were captured near Chestertown, Maryland (USA) from 15 February to 21 March, and 7 October to 13 November 1986. Deer were immobilized by intramuscular injection of 1.1 to 2.2 mg/kg xylazine hydrochloride and 1.8 to 4.4 mg/kg ketamine hydrochloride. Four captive deer from The Pennsylvania State University, Pennsylvania (USA), were immobilized on 16 September 1986 with 1.5 to 2.0 mg/kg xylazine hydrochloride. Intramuscular injection of yohimbine hydrochloride (0.4 mg/kg) was used to antagonize the immobilizations. Free-ranging adult ( > or = 17 months) males could stand after a mean (+/-SE) time of 7.3 +/- 2.4 min, adult females after 8.6 +/- 1.7 min, male fawns after 5.7 +/- 3.3 min, and female fawns after 8.9 +/- 1.9 min. Captive adult males could stand after 20.2 +/- 3.4 min. Intramuscular injections of yohimbine hydrochloride effectively and safely antagonized the xylazine hydrochloride in immobilized deer and were easier to administer than intravenous injections. PMID- 8722290 TI - Cluster of rabies cases of probable bat origin among red foxes in Prince Edward Island, Canada. AB - Between 15 November and 13 December 1993, three cases of rabies of probable bat origin were confirmed in red foxes (Vulpes vulpes) from the same area of Prince Edward Island, Canada, previously thought to be free of rabies in terrestrial mammals. Such clusters have rarely been described in North America. PMID- 8722291 TI - Synergistic inhibitory effect of enviroxime and disoxaril on poliovirus type 1 replication. AB - The effects of enviroxime, disoxaril and ribavirin in pair combinations on poliovirus type 1 (Mahoney) replication in FL cells were tested. Beforehand, the fifty percent inhibitory concentration (IC50) was determined for each compound alone: enviroxime - 0.2 mumol/1, disoxaril - 0.3 mumol/1, ribavirin - 3 mumol/1. Combining enviroxime with disoxaril resulted in synergistic interaction, while combinations with ribavirin were markedly antagonistic. Enviroxime-and disoxaril resistant poliovirus mutants appeared following 10 and 2 consecutive passages in FL cells, respectively. No cross-resistance was observed between these mutants towards disoxaril and enviroxime, respectively. PMID- 8722292 TI - Immunogenicity and protective ability of corpuscular and soluble vaccines prepared from different Coxiella burnetii phase I strains. AB - BALB/c mice immunized intraperitoneally (ip) with killed purified Coxiella burnetii phase I corpuscular vaccines or trichloroacetic acid (TCA) extracts from phase I corpuscles (soluble vaccines) were protected against ip challenge with both homologous and heterologous C. burnetii phase I strains. Though the degree of protection, namely the inhibition of C. burnetii multiplication in the mouse spleen slightly varied, in general, corpuscular vaccines provided better protection than soluble ones. Cross-protection was accompanied by comparable levels of cell-mediated immune response as evaluated by lymphocyte transformation test (LTT). However, higher stimulation indices of LTT were obtained with homologous than with heterologous strains. The values of antibody response as determined by enzyme-linked immunosorbent assay (ELISA) were higher with homologous strains too. On average, both antibody-inducing and antibody-binding capabilities of the strains Priscilla and S were lower than those of the Nine Mile and Luga strains, except for values obtained with the antigens from homologous strains. PMID- 8722293 TI - Antigenic similarity of central European encephalitis and louping-ill viruses. AB - Twenty isolates of Central European encephalitis (CEE) virus were compared with 20 isolates of louping-ill (LI) virus in indirect immunofluorescence test (IIFT), using a panel of 17 monoclonal antibodies (MoAbs) prepared against the prototype LI virus. Three Asian members of the tick-borne encephalitis (TBE) complex were also included in the comparison: Turkish sheep encephalitis (TSE), Russian spring summer encephalitis (RSSE) and Langat (LGT) viruses. Antigenic relationships of the viruses were evaluated by Dice similarity coefficient and cluster analysis. The results revealed antigenic heterogeneity of LI isolates, antigenic homogeneity of CEE isolates, and indicated that CEE and LI are related varieties of Eurasian TBE flavivirus that also includes TSE and RSSE strains. PMID- 8722294 TI - Electropherotypes of rotavirus strains causing gastroenteritis in infants and young children in Tirana, Albania, from 1988 to 1991. AB - During 1988-1991, an epidemiological survey was conducted in Tirana (Albania) on group A rotavirus strains which cause gastroenteritis in infants and young children. Rotaviruses were detected in 312 of 1,241 (25.1%) examined specimens from children with acute diarrhoea. Viruses were detected throughout the study period. Among the 72 rotavirus strains tested for double-stranded RNA (dsRNA) electrophoretic migration pattern, 9 different electropherotypes were recognized, 1 of those being more frequent than the others. At the beginning and at the end of the examined period (1988 and 1990-1991) two different long electropherotypes were predominant, whereas in 1989 (middle period) short electropherotypes were common indicating an involvement of virus strains with short electropherotypes in hospitalization-requiring diarrhoeas occurring in the area surveyed in that year. PMID- 8722295 TI - Detection of spotted fever group rickettsiae in ticks and rodents by polymerase chain reaction technique in People's Republic of China. AB - The polymerase chain reaction (PCR) technique for amplification of genomic fragments of spotted fever group (SFG) rickettsiae directly from field samples of ticks, tick ova, tick larvae, tick faeces and organs of wild mice was employed for the first time in P.R. of China. Ticks and rodents were collected in Beijing and Heilongjiang, Hainan and Hebei Provinces. The PCR primers were designed from the DNA sequence encoding the 190 K protein of R. rickettsii for a 532 bp long product. Seven of ten tick samples, three of four tick ovum samples, one of two tick larva samples, four of seven tick faeces samples (the samples represented pools of several individuals), and two of twenty-seven wild mouse organs were found PCR-positive. In comparison with PCR assay, the haemolymph test gave similar but not so clear-cut results. PCR assay is recommended as a rapid, sensitive and convenient tool for the detection of SFG rickettsiae in endemic foci. The fact that tick faeces were found to certain extent PCR-positive for the presence of SFG rickettsiae is apparently the first report on this subject and contributes to the knowledge of the transmission of these micro-organisms in the nature. PMID- 8722296 TI - Coxiella burnetii in polymorphic lymphocytes in tissue and blood of patients with polymorphic reticulosis. AB - Coxiella burnetii is a well-known causative agent of granulomatous inflammation and an inducer of morphological changes and transformation of human B lymphocytes in vitro. An association of the organism with polymorphic reticulosis (PMR), a malignant granulomatous inflammation characterized by polymorphic lymphocytes, was examined. The infection of C. burnetii was demonstrated in all cases tested, especially in polymorphic lymphocytes. Also the presence of morphologically transformed peripheral blood lymphocytes (PBLs) infected with C. burnetii was demonstrated. In cultures of blood lymphocytes, C. burnetii-infected polymorphic cells identified as B cells became immortalized in vitro. These findings implicate the role of C. burnetii in the process of PMR. PMID- 8722297 TI - Effect of hydrocarbons and crude oil contamination on the sensitivity of French bean to alfalfa mosaic virus. AB - Determination of local necrotic lesions on primary leaves infected by alfalfa mosaic virus (AMV) revealed that hydrocarbons (HC) contamination of the substrate used for cultivation of French bean (Phaseolus vulgaris L., cv. Black Turtle Soup) caused a reduction of bean leaf area and an increase of plant sensitivity to AMV infection. On the other hand, superficial contamination of the leaves by crude oil caused an inhibition of lesion formation. Changes of SDS-polyacrylamide gel electrophoresis (SDS-PAGE) patterns of extractable bean leaf proteins related to the cultivation substrate contamination by HC were also detected. PMID- 8722298 TI - Changes of morphological, biological and antigenic properties of avian influenza A virus haemagglutinin H2 in the course of adaptation to new host. AB - The alterations of avian influenza A virus haemagglutinin (HA) H2 as a result of adaptation to mice were first investigated in this study. HA of mouse-adapted (MA) variant was somewhat different from that of the original strain in electrophoretical mobility, antigenic structure and in haemagglutination activity with mouse red blood cells. PMID- 8722299 TI - Susceptibility of some pea cultivars to pea seed-borne mosaic virus infection and virus transmission by seeds. AB - The susceptibility of 19 pea cultivars to pea seed-borne mosaic virus (PSbMV) by mechanical inoculation and the virus transmission by seeds were determined. All tested cultivars were highly susceptible to the virus infection. The virus transmission by seeds was found in 1.9% - 32.7%. A correlation between mechanical and vector-mediated infections of pea and virus transmission by seeds is discussed. PMID- 8722300 TI - Nutrient composition for cultivation of Japanese encephalitis virus in vitro. AB - Effects of certain groups of nutrients such as glucose, essential amino acids (AA), nonessential AA, vitamins and trace nutrients on the multiplication of various strains of Japanese encephalitis virus (JEV) were studied with an aim to optimise the conditions for cultivation of the virus in porcine stable (PS) kidney cell cultures. Eagle's Minimal Essential Medium (MEM) was modified by addition of the nutrients in different concentrations and combinations. Glucose was found the most important single nutrient in promoting significantly the virus multiplication. Essential AA alone did not influence the virus yield, while in combination with glucose they caused its marked increase. Vitamins and other nutrients did not stimulate significantly virus multiplication. The study revealed that the extent of the glucose effect depends on the virus strain used. PMID- 8722301 TI - Rats with hypertension induced by in utero exposure to maternal low-protein diets fail to increase blood pressure in response to a high salt intake. AB - Hypertension in the rat has been demonstrated to be determined in utero by exposure to maternal low-protein diets. Assessment was made of the response of rats with maternal diet-induced hypertension to a chronic high intake of sodium chloride. Normotensive and hypertensive animals were provided with either drinking water (control) or 1.5% sodium chloride over a 7-day period. Normotensive rats significantly increased blood pressure in response to the increased salt intake. 5-7 days after the start of the study systolic blood pressure was 30-41 mm Hg higher than in controls. 7 days of salt drinking did not alter blood pressure in maternal low-protein diet-exposed, hypertensive rats. In both normotensive and hypertensive groups provision of 1.5% sodium chloride significantly increased fluid intake. In hypertensive rats in the increased fluid intake appeared to be an immediate response, whilst normotensive rats increased intake only after the first day. Urinary volume was increased in both groups of animals receiving salt, within 1 day, and Na+ excretion similarly increased by between 3.5- and 4.5-fold in both groups. K+ excretion initially decreased significantly in both normotensive and hypertensive rats drinking 1.5% sodium chloride, but returned to pre-salt drinking levels by day 5 of the experiment. Rats with maternal-diet-induced hypertension appear to be insensitive to the hypertensive effects of sodium chloride. This insensitivity does not appear to stem from a more rapid clearance of excess sodium, and may relate to other aspects of kidney function and metabolism. PMID- 8722302 TI - Energy expenditure and substrate oxidation rates of obese rats during a 12-day treatment with dexfenfluramine. AB - Serotoninergic neuronal networks are included in regulation and modification of eating behavior and energy metabolism. Dexfenfluramine (dF), a serotonin releaser and reuptake inhibitor, was used to investigate changes in food intake, body weight development, energy expenditure, respiratory quotient, and substrate oxidation rates for 12 days. Rats which had been made obese by postnatal overfeeding received an energy-controlled mash diet and water ad libitum and were intraperitoneally injected with either saline or 5 or 10 mg dF/kg. As compared with controls, food intake and energy expenditure were significantly decreased in a dose-dependent manner, especially during the first 6 days. Lipid oxidation was increased, while the oxidation of carbohydrates was decreased. The body weight was only slightly reduced after 2 days of dF treatment. After 4 days, dF-treated rats resumed body weight, but as compared with controls both dF groups exhibited lower body weights at the end of the experiment. After 12 days the plasma glucose concentration was unchanged, whereas plasma free fatty acids were significantly decreased. Plasma insulin levels were unchanged after dF, but 10 mg dF/kg led to increased muscle and, especially liver glycogen contents, indicating an improved nonoxidative glucose disposal. Muscle pyruvate kinase was slightly but not significantly increased after dF treatment but that of the liver was significantly decreased, indicating a reduced glycolytic activity of the liver. Whereas the renal N excretion was rather decreased, the plasma concentrations of urea, citrulline, arginine, and ornithine were increased, and the liver contents of glutamine and arginine were decreased. Possibly, there is a shift of ammonia removal from glutamine synthesis to production of urea. The sum of all large neutral amino acids in muscle was significantly decreased after dF treatment, indicating a diminished proteolysis. Pair-feeding experiments over 2 days revealed that this was not solely a result of diminished food intake, but also an additional metabolic effect of dF, different from its anorectic effect. It is concluded that both increased oxidation of endogenous fat and reduced food intake could mediate the body weight reducing effect of dF. PMID- 8722303 TI - Vitamin status of healthy subjects in Burgundy (France). AB - A nutrition survey was conducted in Burgundy (France) with a population sample of 337 middle-aged and healthy subjects (157 males and 180 females) recruited at a health examination center in 1985-1986. The status of beta-carotene, thiamin, riboflavin, folate, vitamin B6, B12, C, A, D, and E was assessed by means of 7 day food records and biochemical studies. Results were compared with two other recent nutritional surveys conducted in France: ESVITAF (control group only) and Val de Marne surveys. The dietary information collected for each subject was compared to the 1992 French Recommended Dietary Allowances (FRDA). Dietary vitamin intakes were higher in males than in females. Low vitamin intakes (< 1/2 FRDA) were found in 5% of males and 7% of females for thiamin, in 11% of males and 28% of females for vitamin B6, in 6% of males and 3% of females for vitamin C, in 87% of males and 91% of females for vitamin D, and in 8% of males and 13% of females for vitamin E. No subject had a vitamin intake < 1/2 FRDA for riboflavin, folate, vitamins A and B12. ESVITAF and Val de Marne studies also show low vitamin intakes for vitamin B6, thiamin, riboflavin, vitamins D and E. Biochemical status was examined using erythrocyte enzyme function and blood vitamin levels. The percent of subjects with deficient biochemical values was high for vitamin B6 (15% of all males and 20% of all females), and vitamin D (13% of all males, and 15% of all females). With regard to thiamin, riboflavin, vitamin C, folate, vitamin B12, vitamin A, and vitamin E, < 5% of subjects had values in the range of major vitamin deficiency. However, in both genders, except for vitamin C and vitamin A (only for females), low values corresponding to a moderate risk of vitamin deficiency was high for most vitamins. The incidence of a severe deficient vitamin status for thiamin and riboflavin was higher in Val de Marne than in Burgundy, or ESVITAF. In Val de Marne, the probability of a moderate risk of vitamin deficiency was high for thiamin, riboflavin, vitamin B6, vitamin A and vitamin E. Our results (as other studies performed in France and in other industrialized countries) raise the issue of the health significance of marginally deficient vitamin status. PMID- 8722304 TI - Liver transglutaminases and vitamin-A deficiency in hairless mice. AB - In hairless mice, a moderate vitamin-A deficiency, without any clinical signs or weight changes, reduces the activity of soluble cytoplasmic hepatic transglutaminase without affecting the membrane form of the enzyme. This attack of soluble transglutaminase appears to be a biological marker of early deficiency. The relations between this disturbance and the hepatocyte sensitivity to aggressors at this stage are discussed. PMID- 8722305 TI - Characterisation and reactivity of a glycidic antigenic component in Toxoplasma gondii. AB - Toxoplasma gondii is composed of a complex antigenic mosaic and some antigenic fractions are of major diagnostic interest. We studied an antigenic fraction with molecular weight < 17 kda, identified by SDS-PAGE 15% and immunoblotting. This fraction could be easily extracted from the whole antigen with phenol/CH-Cl3 and its analysis showed a mainly carbohydrate composition. With immunoblotting this fraction showed strong reactivity with specific IgM and IgG in early-acute infection; IgM reactivity disappears or is much less marked in chronic acquired infection. Extraction with phenol/CHCl3 allows this fraction to be used as an antigen in other diagnostic methods. PMID- 8722306 TI - Leishmania donovani lipophosphoglycan (LPG) inhibits respiratory burst and chemotaxis of dog phagocytes. AB - Lipophosphoglycan (LPG) is the major glycoconjugate of Leishmania promastigote surface membrane. Previous studies on human and murine models have demonstrated that this molecule is involved in the attachment and survival of Leishmania in the host cells. Dog is the main reservoir of Leishmania strains responsible for human leishmaniasis in Italy. Since no studies have been performed on the LPG canine phagocyte interactions, we investigated the LPG effects on dog phagocyte functions by evaluating: 1) the chemotactic activity of peripheral monocytes and polymorphonuclear (PMN) cells, in terms of cellular polarization; 2) the PMN cell respiratory burst, by measuring superoxide anion and hydrogen peroxide production. Results demonstrated a significant reduction of metabolic and chemotactic activity in LPG-preincubated cells, thus emphasizing the ability of this molecule to impair also the canine phagocyte responses. PMID- 8722307 TI - Characterisation of a phospholipase C produced by Pseudomonas fluorescens. AB - Phospholipase C (phosphatidylcholine phosphohydrolase, EC 3.1.4.3) and lipase (EC 3.1.1.3) activities were detected in the supernatant fluid of Pseudomonas fluorescens strain D cultures. A combination of ultrafiltration and successive chromatography through columns of Sephadex G-75 and DEAE-cellulose was used to purify the phospholipase C over 700-fold from the culture medium, with 28.5% yield. The purified enzyme appeared as a single band after polyacrylamide gel electrophoresis. The apparent molecular mass of the phospholipase C was 36,000 daltons when estimated by gel permeation chromatography. The purified enzyme hydrolysed phosphatidylcholine more efficiently than phosphatidylethanolamine. The synthetic substrate p-nitrophenylphosphorylcholine, phosphatidylinositol or sphingomyelin were not hydrolysed. Hydrolysis of phosphatidylcholine was inhibited by EDTA (1mM) and stimulated by Zn2+, Mg2+ ions and detergents. These properties of the enzyme indicate that it is distinct from the previously reported Ps. fluorescens phospholipase C. PMID- 8722308 TI - Composition of the Lactobacillus acidophilus complex isolated from vaginal flora. AB - One hundred and fifty human vaginal samples containing a diversity of pathogens or nonpathogens (Gardnerella vaginalis, Streptococcus sp., Staphylococcus sp., Candida albicans. Mycoplasma sp.) were examined for their content in lactobacilli of the Lactobacillus acidophilus complex. Although all samples contained lactobacilli, strains of the L. acidophilus complex were present in only twenty nine cases. Isolates were further characterized and compared with type strains or reference strains in an attempt to differentiate by phenotypic means the genospecies of the L. acidophilus complex. Data regarding specific activities of beta-galactosidase (beta-gal) and of phospho-beta-galactosidase (P-beta-gal) provided no specific information at the species level within the L. acidophilus complex. DNA-relatedness differentiates this genospecies. Most lactobacilli isolated from the vaginal flora of symptomatic women were genotypically close to L. gasseri CIP 102991T by the technique of DNA/DNA hybridization. PMID- 8722309 TI - Effect of sucrose consumption on level of Streptococcus mutans in saliva. AB - The aim of the present paper was to study whether sucrose consumption may affect the level of salivary Streptococcus mutans. The study was justified by the disagreement between results of "in vitro", animal and human experiments and results of epidemiological studies made on study-populations. According to experimental research, sucrose availability promotes Streptococcus mutans selection and development in the oral cavity. On the other hand, the results of epidemiological studies run counter to these experiments. In the present study, the main reasons for this disagreement were investigated and identified. From this, an epidemiological study was performed, taking into account these potentially confounding factors. The results show that sucrose consumption is actually able to promote Streptococcus mutans selection and development in the oral cavity, thus increasing the risk of dental caries. PMID- 8722310 TI - Rapid diagnosis of influenza type A infection: comparison of shell-vial culture, directigen flu-A and enzyme-linked immunosorbent assay. AB - Two direct antigen detection methods (Directigen FLU A, Becton-Dickinson, Cokeysville, Md. and Influenza A virus ELISA Antigen detection kit, Alpha Biotech, Milano) were compared with isolation by shell-vial cultures for rapid diagnosis of influenza type A virus infection. The three methods were performed using a reference viral suspension consisting of a clinical isolate of influenza type A virus as well as 41 nasopharyngeal aspirates collected from patients with influenza symptoms. Influenza type A virus was recovered from 14 specimens by shell-vials. Ten specimens were positive by Directigen FLU A and 7 by ELISA antigen detection kit. When performed on clinical samples the sensitivity of Directigen FLU A and ELISA was 64.2% and 42.8%, respectively. The limit of sensitivity of the shell-vials, performed on a virus stock suspension titrated by haemagglutinin activity, was 0.06-0.03 HA units/ml at 24 hours or 72 hours after inoculation. Directigen FLU A and ELISA detected as positive a virus stock suspension containing 0.25 HA units/ml and 1 HA units/ml, respectively. PMID- 8722311 TI - Gastric infection by Helicobacter pylori and antral gastritis in hyperglycemic obese and in diabetic subjects. AB - The aim of this work was to ascertain if diabetes and obesity can affect gastric colonization by Helicobacter pylori. 59 hospitalized subjects with dyspepsia and endoscopic antral gastritis were selected. They were divided into three groups: I) 13 patients with normal body weight and without disease other than gastritis (control group); II) 15 patients with essential obesity of whom 10 had impaired glucose tolerance (IGT); III) 31 patients with type II diabetes mellitus, of whom 14 were obese. Three gastric biopsies were obtained from each patient for histologic examination and H. pylori detection by means of rapid urea test, culture and histological evidence of Helicobacter-Like Organisms (HLO). Age, sex, blood glucose, cholesterol, triglycerides, HDL-cholesterol, basal gastrine, duration of illness, body weight were statistically analysed. Differences between the three groups were not statistically significant. There was a higher prevalence of H. pylori infection both in obese and in diabetic patients with respect to control subjects. Prevalence became still higher in obese patients with impaired glucose tolerance. Among the three tests used for the detection of H. pylori, culture and rapid urea were reliable and specific, while the histologic test was highly sensitive but barely specific. Our data suggest that both obesity and type II diabetes may be associated with an increased incidence of H. pylori-colonization. This could be related to the reduced gastric motility observed in both pathologies and chemical changes in gastric mucosa following non enzymatic glycosylation processes. PMID- 8722312 TI - Distribution and numerical taxonomy of Vibrionaceae in the waters of the Straits of Messina. AB - The results of a study carried out by numerical analysis on Vibrio strains isolated from the waters of the Straits of Messina are reported. The quantitative data showed the presence of low bacterial densities (ranging from 9 to 99 CFU/100 ml of water) due to the intense currents which characterize this area; also the highest bacterial counts were generally found during the "montante" current. With regard to the qualitative results, there was a predominance of vibrios belonging to V. mediterranei, V. splendidus II and V. pelagius II species, which represented respectively 25%, 19% and 13% of the total bacterial population. The species distribution did not seem related to sampling stations or depth. The taxonomic structure, obtained using the simple matching coefficient and unweighted average linkage clustering, revealed the presence of 7 main clusters (S = 80-95%), which included strains of various origin because of the particular turbulence of the waters of the Straits. PMID- 8722313 TI - Absent inhibition of hepatitis B virus replication by azidothymidine in patients chronically coinfected by HIV-1 and HBV. AB - The effect of Azidothymidine (AZT) in vivo on the replication of the Hepatitis B Virus (HBV) was studied in a population of 25 patients chronically coinfected by HBV and Human Immunodeficiency Virus Type 1 (HIV-1), and receiving AZT at the usual dosage. The drug effect was evaluated by sequential measurement of the HBV DNA level. No significant activity at short and medium term was found on HBV replication in either homosexuals or IV drug users chronically coinfected by HIV 1 and HBV. PMID- 8722314 TI - Antibodies to Borrelia burgdorferi in sheep and goats. Alto Adige-South Tyrol, Italy. AB - A serologic survey for antibodies to Borrelia burgdorferi was conducted on sheep and goat serum samples collected in Alto Adige-South Tyrol, Italy, in 1990. Sera were tested by Indirect Immune Fluorescence Assay (IIFA) and Microbial Adherence Immobilization Assay (MAIA). IIFA and/or MAIA anti-B. burgdorferi antibodies were detected in 14.1% of the 269 sheep and 36.8% of the 133 goats examined. IIFA and MAIA were both positive in 4 out of 38 positive sheep sera (10.5%) and 21 out of 49 positive goat sera (42.8%). These discrepancies suggest that MAIA- and IIFA detected antibodies do differ from each other. The detection by MAIA of antibodies sensitizing B. burgdorferi to the killing effect of complement seems to be a valid parameter to evaluate the acquired immunity of sheep and goats to B. burgdorferi infections. PMID- 8722315 TI - Leptospira interrogans serovar hardjo in the kidneys and genital tracts of naturally infected sheep. AB - A bacteriological study was carried out to identify possible renal and/or genital carriers of Leptospira interrogans serovar hardjo. L. hardjo was found at slaughter in the kidneys of three seropositive ewes, but not in uterus or salpinges of these animals. PMID- 8722316 TI - Preparation of serotype specific antiserum against porcine adenoviruses in mice. AB - Serotyping of porcine adenoviruses needs type-specific antiserum. The injections of purified virions to SPF Balb/C mice, followed by an intraperitoneal injection of Ehrlich ascitic tumour cells, gave enough ascitic fluid containing practically applicable type-specific antibody. PMID- 8722317 TI - Role of endocrine cell microvesicles in intercellular chemical transduction. AB - Microvesicles (MVs) in endocrine cells are morphologically similar to neuronal synaptic vesicles. MVs were shown to contain proteins involved in neurotransmitter storage such as vacuolar H(+)-ATPase and neurotransmitter transporters, and ones in vesicular trafficking such as synaptobrevins and N ethylmaleimide-sensitive fusion protein. Isolated MVs accumulate cell-specific neurotransmitters in an energy-dependent manner. Upon stimulation, the MVs may fuse with the plasma membrane and secrete the internal neurotransmitters. Thus, endocrine cells possess an MV-mediated secretion system as an intercellular signal transducing system. PMID- 8722318 TI - Properties of particulate transglutaminase from Yoshida tumor cells. AB - Homogenates of Yoshida hepatoma cells, cultured as ascite suspension in vivo, display significant transglutaminase activity in both the cytosolic and the particulate fraction. The enzyme, however, is predominantly membrane-bound. Transglutaminase was solubilized from the membranes either by extraction with detergents or treatment with neutralized hydroxylamine or proteinases. We observed similar molecular weight under denaturing conditions, catalytic and immunologic properties for purified cytosolic and solubilized transglutaminase, and identity of the limited proteolytic maps. These results suggest that transglutaminase isoforms actually consist of the same protein undergoing translocation by unknown mechanisms. PMID- 8722319 TI - Structure of recombinant human parathyroid hormone in solution using multidimensional NMR spectroscopy. AB - The solution structure of human parathyroid hormone, in the form of recombinant prolyl-hPTH(1-84), has been investigated by multidimensional NMR spectroscopy under conditions (aqueous trifluoroethanol) which favour the structured-state of the protein. Spin systems were identified from 3D 1H DQF (double-quantum filtered)-COSY and TOCSY spectra and sequence-specific assignments were from 2D 1H phase-sensitive NOESY spectra. Signal overlap was resolved in a 3D-NOESY-TOCSY spectrum and assignments were confirmed with 2D NOESY-15N-HMQC (heteronuclear multiple-quantum coherence) spectra taken of a sample universally labeled with 15N. A satisfactory set of final structures was calculated from the quantitative NOE data using restrained molecular dynamics and energy minimization calculations. The N-terminus is dominated by three, well defined helices between Ser-3 to Asn-10, Ser-17 to Lys-27 and Asp-30 to Leu-37, while the most significant structural features in the C-terminus are a short, less-well defined helix between Asn-57 to Ser-62 and a series of loose turns. These two terminal units are joined by an unstructured mid-region. The molecule shows a tendency towards tertiary structure, defined by a number of long-range NOEs. A detailed RMS deviation analysis allowed the final refined structures to be classified into a limited ensemble of stable conformations that reflect the inherent flexibility of the hormone in solution. PMID- 8722320 TI - Evaluation of HBV promoters for use in hepatic gene therapy. AB - Strategies for in vivo hepatic gene therapy will require regulatory elements which allow for long-term expression of therapeutic genes and restriction of expression to hepatocytes. This study investigates the suitability of promoters derived from hepatitis B virus (HBV) for liver-specific gene expression in vectors for hepatic gene therapy. We provide three hepatocyte-specific promoters, the HBV core promoter, the HBV core promoter linked directly to the HBV enhancer I, and a hybrid promoter containing the HBV enhancer II and a basic CMV promoter, which are hepatocyte-specific and allow for increasing levels of reporter gene expression. Moreover, in long-term expression studies using our promoter constructs in the context of an EBV based expression system we found that expression from these promoters remained nearly unchanged over a period of at least two months in hepatocyte-derived cell lines. PMID- 8722321 TI - Inactivation of the very strong HCMV immediate early promoter by DNA CpG methylation in vitro. AB - The influence of DNA methylation in vitro on the activity of the very strong human cytomegalovirus (HCMV) major immediate early (IE) modulator/enhancer/promoter region was investigated by transient transfection experiments of premonocytic HL-60 cells. While sequence-specific methylation of the major IE enhancer and/or modulator with the cytosine methyl-transferases FnuDII, HhaI and HaeIII had no significant effect, the promoter activity was completely repressed by methylation of the cytosine in 5'-CpG sites with the Spiroplasma methyltransferase SssI. Addition of TNF-alpha or PMA which are strong stimulators of HCMV major IE enhancer/promoter activity in premonocytic HL-60 cells had no effect on repression. Inactivation of the IE enhancer/promoter via methylation by M.SssI could be partially alleviated by co-transfection with an excess of untranscribable highly methylated DNA. These results indicate that a methyl-CpG binding factor is involved as mediator in the inhibitory effect of HCMV enhancer/promoter methylation. Taken together, the HCMV major IE enhancer/ promoter has been shown to be susceptible to transcriptional inactivation by methylation of the cytosines in CpG dinucleotides, a process that is proposed to play a modulatory role in viral latency. PMID- 8722322 TI - Comparison of the substrate specificities of cAMP-dependent protein kinase from bovine heart and Ascaris suum muscle. AB - The catalytic subunits of cAMP-dependent protein kinases (protein kinase A) from bovine heart and Ascaris suum muscle exhibit only 48% sequence identity and show quantitative differences in substrate specificity. These differences were not obvious at the level of short synthetic substrate peptides but were distinct for some protein substrates. Phosphofructokinase from Ascaris, a physiological substrate, was a better substrate for the protein kinase from the nematode in comparison to the mammalian protein kinase due to a 10-fold lower Michaelis constant. Selective phosphorylation by the two kinases was also observed with some in vitro substrates. In addition, quantitative differences in the interactions between R- and C-subunits from Ascaris and bovine heart were observed. However, several synthetic peptides whose sequence reflected the phosphorylation site of Ascaris suum phosphofructokinase (AKGRSDS*IV), or variations of it, were phosphorylated with the same efficiency by both protein kinases. Based on the data the following are concluded: (1) In agreement with the conservation of structure in the catalytic cleft, the recognition of substrates by protein kinases from phylogenetically distant organisms exhibits similarity. (2) Non-conserved parts of the surface of the protein kinase molecule may contribute to binding of protein substrates and thus to selective recognition. PMID- 8722323 TI - Heterologous expression and characterisation of mouse brain fatty acid binding protein. AB - A novel brain-type member of the fatty acid binding protein family (B-FABP) was heterologously expressed in Escherichia coli, either as inclusion bodies at 37 degrees C or in soluble form at 22 degrees C. Both B-FABP renatured from inclusion bodies and the solubly expressed protein could be purified to homogeneity by anion exchange chromatography and gel filtration in a functional conformation as they bound oleic acid with high affinity. None of the five cysteines of B-FABP was involved in disulphide bond formation. Isoelectric focusing revealed heterogeneity of the renatured protein but not of the solubly expressed protein. By Western blotting using affinity purified rabbit antibodies raised against the recombinant B-FABP it was demonstrated that in adult mice, B FABP is predominantly expressed in the olfactory bulb. PMID- 8722324 TI - First international symposium on allogeneic peripheral blood precursor cell transplants. PMID- 8722325 TI - The evolution of clinical peripheral blood stem cell transplantation. PMID- 8722326 TI - Biological characteristics of cells migrating into the bloodstream. PMID- 8722327 TI - Allogeneic peripheral blood precursor cell transplants in rabbits. AB - This report summarizes a series of experiments undertaken to evaluate the role of mobilized peripheral blood precursor cells (PBPC) for transplantation across a major histocompatibility barrier. Adult outbred red Burgundy rabbits were used as donors, New Zealand white rabbits of the opposite sex as recipients. Conditioning consisted of single dose total body irradiation (TBI) of 10 Gy supported by a short course of cyclosporine to enhance engraftment. Human recombinant G-CSF at a dose of 10 micrograms/kg was used for mobilization of precursor cells. Three methods of PBPC transplants were tested initially in 5 animals each. PBPC were collected and infused at once on day 0; collected initially, cryopreserved for one month, infused on day 0 and followed by 3 additional fresh donations or collected and infused on 6 occasions between days 0 and + 11. 13 animals engrafted, 2 became complete, longterm chimeras. Survival was best in the group given repetitive infusions (39 days median, 12 days to > 180 days, range). 10 additional animals were transplanted as in the last group and the number of transplanted nucleated cells (10.5 x 10(8)/kg median, 7.3 - 15.7 x 10(8)/kg range) and colony forming units CFU-GM (42 x 10(4)/kg median, 12.3 - 176.8 x 10(4)/kg range), were compared with outcome. Median survival of the 10 animals was 29 days (12 - 55 days range; 1 autologous reconstitution). Survival did not correlate with total nucleated cells per kg (r = 0.10; p = 0.79), but there was a trend to prolong survival with higher numbers of CFU-GM per kg (r = 0.47; p = 0.19). These data show that allogeneic PBPCT can engraft across a major histocompatibility barrier, that the high number of CFU-GM per kg might be advantageous, but also that additional methods are warranted to reduce acute GvHD. PMID- 8722328 TI - Treatment of normal donors with recombinant growth factors for transplantation of allogeneic blood stem cells. PMID- 8722329 TI - PBPC grafts from healthy donors: analysis of CD34+ and CD3+ subpopulations. PMID- 8722330 TI - Safety of AlloPBPCT donors: biometrical considerations on monitoring long term risks. AB - Up to now there are no data on long-term effects of allogeneic peripheral blood cell transplantation (AlloPBPCT). In particular, long term effects on healthy donors by the mobilization procedure which includes the exposition to G-CSF over several days are unknown. Recently the possibility of an increase in risk for acute leukaemia in this cohort has been discussed. Systematic long-term safety monitoring for AlloPBPCT donors cannot be adequately planned without agreeing on a both relevant and reasonably pessimistic hypothetical size of the increased leukaemia risk to be detected if present. Using data on leukaemia after treatment for Hodgkin's disease as example it is argued that a) excess leukaemia cases should be expected to occur predominantly between 2 and 10 years after the leukaemogenic event and b) a reasonably pessimistic guess would expect about 0.5% leukaemia cases at 10 years in AlloPBPCT donors. Such a tenfold increase over the general population's 10 year leukaemia incidence would be relevant, but require long-term follow up of several thousands of donors to demonstrate or exclude. In conclusion, safety monitoring for AlloPBPCT donors can only be organized on an international scale. PMID- 8722331 TI - Enrichment of allogeneic CD34+ cells and T cell depletion by percoll density gradient centrifugation. PMID- 8722332 TI - Mobilization and purification of CD34+ cells from normal donors-regimens with G CSF, GM-CSF, or a combination of both. PMID- 8722333 TI - Transplantation of allogeneic CD34+ peripheral blood stem cells (PBSC) in patients with advanced hematologic malignancy. PMID- 8722334 TI - Transplantation of allogeneic peripheral blood progenitor cells--the EBMT experience. PMID- 8722335 TI - Clinical results of allogeneic peripheral blood stem cell transplantation: Japanese survey 1995. PMID- 8722336 TI - Allogeneic peripheral blood progenitor cell transplantation--the Australian experience. PMID- 8722337 TI - Allogeneic transplantation for hematological malignancies using rho G-CSF mobilized blood stem cells. PMID- 8722338 TI - Transplantation of allogeneic peripheral blood stem cells. PMID- 8722339 TI - Addition of PBPCs to the marrow inoculum allows engraftment of mismatched T cell depleted transplants for acute leukemia. PMID- 8722340 TI - Transplantation of peripheral blood progenitor cells from unrelated donors. AB - Six patients with high risk haematological malignancies received peripheral blood progenitor cells (PBPC) from unrelated donors. Four patients received PBPC as primary treatment and 2 following graft failure. Five donors were HLA-A, -B and DR identical and one had a one antigen mismatch. PBPC were mobilised by treatment with G-CSF for 4-6 days. The patients received a range of 3.4 to 11.4 x 10(8) mononuclear cells/kg and 1.0 to 15.0 x 10(6) CD34 positive cells/kg. Four patients were given Campath 1G and 2 ATG prior to transplantation. The patient with one antigen mismatch received in vitro T-cell depleted PBPC using Campath 1M. All received cyclosporin and 5 in addition methotrexate. All recipients were given G-CSF and all engrafted. The patients developed no or mild acute GVHD. Two patients had limited chronic GVHD of the skin. The recipient of the mismatched graft died from extensive chronic GVHD. Three patients have had a relapse and two are alive and free of leukaemia. PMID- 8722341 TI - Allogeneic and autologous transplantation for haematological diseases, solid tumours and immune disorders. Current practice in Europe in 1996 and proposals for an operational classification. Accreditation Sub-Committee of the European Group for Blood and Marrow Transplantation (EBMT). AB - The indications and techniques employed for haematopoietic stem cell transplants performed in Europe differ substantially. The Accreditation Sub-Committee of the EBMT has consulted widely and has attempted to describe practice in Europe current in 1996 and in particular to define whether a particular procedure should be regarded as 'Routine' or whether it should be carried out preferably in the context of a Clinical Research Protocol or as part of a 'Developmental' pilot study. It is recognised that a classification of this kind must be imperfect and will require revision at frequent intervals. PMID- 8722342 TI - Bone marrow engraftment following unrelated donor transplantation utilizing busulfan and cyclophosphamide preparatory chemotherapy. AB - Two major problems of unrelated donor transplantation have been an increased incidence of GVHD and graft failure. Even with HLA identity by microlymphocytotoxicity assay and non-reactive MLC, URD marrow transplant recipients have a higher incidence of graft rejection and GVHD. The preparative regimen busulfan 16 mg/kg and cyclophosphamide 120 mg/kg (BuCy2) has been shown to be at least as effective in preparation of recipients with CML of HLA identical sibling grafts as cyclophosphamide and total body irradiation (Cy/TBI). However, concern about a high rejection rate in URD transplants has prevented most centers from using BuCy2 in this setting. From March 1990 to March 1994, 26 patients underwent URD transplantation following preparation with BuCy2. Patients received either standard cyclosporine and methotrexate or cyclosporine and methylprednisolone for GVHD prophylaxis. Two patients died on day 16 and 20 without evidence of hematopoietic engraftment. Of the 24 patients evaluable for engraftment, 23 (96%) had evidence of donor engraftment defined as an ANC > 0.5 x 10(9)/1. No patient who had initial engraftment had late graft failure. Within our study group the risk of graft rejection or graft failure does not appear to be higher than that reported for URD transplants utilizing TBI-containing regimens. PMID- 8722343 TI - Phase I study of high-dose continuous intravenous infusion of VP-16 in combination with high-dose melphalan followed by autologous bone marrow transplantation in children with stage IV neuroblastoma. AB - The purpose of the study was to determine the maximum tolerated dose of continuous infusion of high-dose VP-16 in combination with high-dose melphalan (HDM) for conditioning before autologous bone marrow transplantation (ABMT). Thirteen children (median age 27 months) with stage IV neuroblastoma were treated with high-dose VP-16 and HDM followed by ABMT as consolidation treatment. All had previously received conventional chemotherapy with a mean number of six drugs. Surgery of the primary tumor had been performed in 12/13. We performed a dose escalating study of VP-16 from 1800 mg/m2/72 h with 300 mg/m2/72 h dose increments according to toxicity. VP-16 was administered as a 72-h i.v. infusion. Melphalan (140 mg/m2/day) was administered once as an i.v. push. VP-16 pharmacokinetics were analyzed in 12 patients. Five children received 1800 mg/m2/72 h of VP-16, five received 2100 mg/m2/72 h and three, 2400 mg/m2/72 h. The mean duration of granulocytopenia (< 0.5 x 10(9)/1) was 24 days and thrombocytopenia (< 50 x 10(9)/1) was 36 days. No major infectious complications occurred. Gastrointestinal (GI) toxicity was the dose-limiting toxicity. Five severe manifestations of GI toxicity in three patients led us to consider 2400 mg/m2/72 h as the MTD. The mean VP-16 clearance rate was 17.3 ml/min/m2 with continuous infusion. A mean steady-state plasma concentration of 24.2 micrograms/ml (s.d. = 2) and 28.3 micrograms/ml (s.d. = 1.9) was achieved at the 1800 mg/ml and 2100 mg/m2 dose levels, respectively, GI toxicity is dose limiting when VP-16 at 2400 mg/m2/72 h, is associated with HDM. When given as a continuous i.v. infusion, at 2100 mg/m2/72 h, VP-16 associated with HDM is well tolerated before ABMT in young heavily pre-treated children. PMID- 8722344 TI - Busulfan, cyclophosphamide and fractionated total body irradiation for autologous or syngeneic marrow transplantation for acute and chronic myelogenous leukemia: phase I dose escalation of busulfan based on targeted plasma levels. AB - In a previous phase I study, it was concluded that tolerable doses of busulfan (BU), cyclophosphamide (CY) and total body irradiation (TBI) were 8 mg/kg, 60 mg/kg and 12.0 Gy, respectively, for autologous marrow transplant recipients. In an attempt to decrease the variability of BU steady-state concentration (Css) following oral dosing, a BU dose escalation study based on targeted plasma levels was performed in patients receiving autologous transplants for AML or syngeneic transplants for CML. In this study, the BU dose was adjusted up or down based on observed plasma concentration. All patients received a fixed dose of CY 60 mg/kg and TBI of 12 Gy. The first dose level evaluated was 8.6 mg/kg with a target BU Css of 511 ng/ml. Eight patients were entered at this level and the median BU Css achieved was 441 (range 253-566). One of eight patients developed grade 3-4 regimen-related toxicities (RRT). The oral dose of BU for dose level II was 10.6 mg/kg with a target Css of 632 ng/ml. Six patients were entered at this level and median BU Css achieved was 642 (range 566-674). One of six patients developed grade 3-4 RRT. The oral dose for dose level III was 12.6 mg/kg with a target BU Css of 754 ng/ml. Five patients with AML were entered at this dose level and the median plasma BU Css was 733 ng/ml (682-900). Two of five (40%) patients at dose level III developed grade 3-4 RRT which was considered excessive making dose level II the MTD. This study showed that targeted BU Css can reliably be achieved with a bias of -5.23% and mean absolute error of 11.3%. Overall, targeting made a -32.5% to 158.3% change in plasma BU Css as compared to expected BU Css based on first dose pharmacokinetics if targeting were not performed in this study. Thus, targeting avoided much of the variability in BU Css seen in other studies and appears to have allowed for an increase in oral dosing from 8 mg/kg to 10.6 mg/kg. Despite achieving higher and more uniform BU Css, there was no apparent effect on relapse or survival, although the number of patients evaluated was small. PMID- 8722345 TI - Urinary elimination of cyclophosphamide alkylating metabolites and free thiols following two administration schedules of high-dose cyclophosphamide and mesna. AB - Twenty patients with a variety of neoplastic diseases were treated with preparative regimens containing high-dose cyclophosphamide (CY) administered as a 2-h infusion (60 mg/kg) for 2 days or by continuous infusion (1500 mg/m2/day) for 4 days. In patients receiving CY by 2-h infusion, the uroprotectant 2 mercaptoethane sulfonate (MESNA) was administered as an intermittent, bolus intravenous infusion (20% of CY dose) every 6 h. In patients receiving continuous infusion CY, MESNA was administrated concomitantly at an equivalent dose to CY by continuous infusion. During the first 24 h of CY administration, urine was collected at 2-h intervals and analyzed for free thiols and CY-alkylating metabolites. In patients receiving CY by short infusion and MESNA by intermittent bolus infusion, urinary concentrations of alkylating metabolites peaked at 4-8 h. During each dose of MESNA, urinary free thiols peaked at 2 h following administration but fell to pre-treatment levels at subsequent intervals. In patients receiving CY by continuous infusion, CY alkylating metabolites increased gradually over the 24-h study period while free thiols remained at a constant level during this period. With bolus administration of CY and intermittent bolus administration of MESNA every 6 h, there are periods where urinary CY-alkylating metabolites are elevated and free thiol concentrations are diminished. During continuous infusion of CY and MESNA, urinary CY alkylating metabolites reached peak concentrations at 18-22 h while the exposure of the bladder to free thiols remained constant. Recommendations are provided to increase the exposure of free thiols in the bladder when MESNA is administered by bolus or continuous infusion. PMID- 8722347 TI - The absolute number of circulating CD34+ cells predicts the number of hematopoietic stem cells that can be collected by apheresis. AB - We report our single institution's effort to standardize the method of collecting peripheral blood progenitor cells (PBPC) used for autologous transplantation. PBPC were mobilized by different types of chemotherapy followed by G-CSF (24 patients) or G-CSF alone (six patients) in 30 patients with various underlying neoplastic diseases. A median of three aphereses (range: 2-7), using the CS3000 cell separator was performed and a blood volume of 101 was processed. We studied the relationship between the absolute numbers of circulating leukocytes, mononuclear cells and CD34+ cells and the amount of PBPC collected during a single apheresis procedure. CD34+ cells were quantified by leukocyte subset analysis based on flow cytometry. Both CFU-GM and CD34+ cell contents of the apheresis products (69 procedures analyzed) correlated strongly: rho = 0.936, P = 0.0001). Regression analysis showed that the total number of CD34+ cells or CFU GM content of the apheresis products could be predicted (r = 0.915, P = 0.0001) from the absolute number of CD34+ cells in the blood. A number of 10 circulating CD34+ cells per mm3 blood ensured a minimum of 0.5 x 10(6) CD34+ cells per kg, collected on the same day. Of the 30 patients, 17 received an autologous graft that contained only PBPC in 13. Long-term and complete hematological reconstitution was observed in all patients who had a minimum of 2 x 10(6) CD34+ cells per kg reinfused. PMID- 8722346 TI - Peripheral blood progenitor cell transplantation: a single centre experience comparing two mobilisation regimens in 67 patients. AB - Between June 1991 and January 1995 we performed 67 peripheral blood progenitor cell transplants (PBPCT). Ten patients (group 1) were mobilised with 7 gm/m2 of cyclophosphamide followed by daily G-CSF injections (5 micrograms/kg, subcutaneously). When the white cell count reached 1 x 10(9)/1 they were leukapheresed for 5 days. After stem cell infusion they received G-CSF (10 micrograms/kg/day) until the neutrophil count reached 1.5 x 10(9)/1. Fifty-six patients had PBPCs mobilised with 3 gm/m2 of cyclophosphamide followed by daily subcutaneous G-CSF (5 micrograms/kg) and PBPCs were harvested on 2 consecutive days, when the white cell count rose to 4 x 10(9)/1. After stem cell infusion this group did not receive G-CSF. In 47 of the 56 patients (group 2) adequate MNC (> or = 4 x 10(8)/kg) and/or CFU-GM (> or = 10 x 10(4)/kg) were obtained. Insufficient MNC and/or CFU-GM were obtained in 10 patients. They were therefore transplanted using a combination of bone marrow and peripheral blood progenitor cells (group 3). Overall 64 patients successfully engrafted. Median days to neutrophils > or = 0.5 x 10(9)/1 were 9 (range 8-13), 12 (range 8-25) and 11 (range 9-16) and to platelets > or = 50 x 10(9)/1 were 11 (range 9-23), 13 (range 9-90) and 16 (range 13-99) in groups 1, 2 and 3 respectively. Patients in group 1 had a faster neutrophil recovery than patients in group 2 (P = 0.0002). The three patients who failed to engraft all received a combination of autologous peripheral blood and bone marrow cells. PMID- 8722348 TI - Density enrichment and characterization of hematopoietic progenitors and stem cells from umbilical cord blood. AB - Umbilical cord blood (UCB) is being used for hematopoietic rescue after myeloablative therapy in a rapidly growing number of patients. Recent developments of cord blood banking, ex vivo progenitor expansion and gene therapy techniques have raised the issue of efficient progenitor and stem cell enrichment procedures using UCB. We have used discontinuous density gradient techniques to analyze progenitor distribution in the mononuclear cell fraction of cord blood. This resulted in establishment of a highly reproducible, rapid, cost-effective single-step density separation method that generates a light density fraction, which when compared to conventional mononuclear cells has a high number of clonogenic progenitors, can be extensively expanded in vitro for up to 21 days and has the ability to sustain long-term hematopoiesis when inoculated on a preformed stromal layer. It can also serve as an efficient target for retrovirally mediated gene transfer, utilizing a vector expressing a mutated dihydrofolate reductase gene that confers methotrexate resistance. PMID- 8722349 TI - Combination therapy with G-CSF and erythropoietin after autologous bone marrow transplantation for lymphoid malignancies: a randomized trial. AB - Previous studies have shown that, unlike in patients submitted to allogeneic BMT, administration of recombinant erythropoietin (Epo) after autologous BMT (ABMT) had no significant effect on erythroid recovery and transfusional requirements. On the other hand, it has also been shown that combining Epo with recombinant granulocyte colony-stimulating factor (G-CSF) in patients with the acquired immunodeficiency syndrome (AIDS) and with myelodysplastic syndromes resulted in additive effects on erythropoiesis. To test the effects of combined G-CSF + Epo therapy on erythroid recovery after autologous bone marrow transplantation a pilot randomized, three-arm trial was designed. Thirty patients suffering from lymphoid malignancies submitted to ABMT were randomly assigned to receive G-CSF alone (5 micrograms/kg, from day + 1 up to reaching an ANC > or = 10(9)/1), G-CSF + Epo (150 U/kg, from day +1 to +21), or neither of these (controls). Patients receiving G-CSF + Epo had significantly more reticulocytes on day +21 and reached 30 x 10(9)/1 reticulocytes earlier when compared to both G-CSF and control patients; however, the number of red blood cell (RBC) transfusions was not modified by the addition of Epo to G-CSF, although both groups had significantly fewer units transfused than controls. No effect on platelet recovery or platelet transfusional requirements was observed. Myeloid recovery was comparable in the G CSF and G-CSF+Epo groups, and significantly accelerated as compared to controls. We conclude that the addition of Epo to G-CSF causes a slight acceleration of erythroid recovery after ABMT, but is not associated with transfusional benefits. Therefore, the present data do not support the use of Epo in association with G CSF to hasten erythroid recovery after ABMT. PMID- 8722350 TI - Administration of G-CSF can be delayed after transplantation of autologous G-CSF primed blood stem cells: a randomized study. AB - Hematopoietic growth factors like G-CSF or GM-CSF have been shown to shorten the period of severe neutropenia after HD chemotherapy and autologous BMT, and are now widely used to mobilize hemopoietic stem cells into peripheral blood. In order to evaluate the possibility of delaying G-CSF administration after transplantation of G-CSF mobilized blood stem cells (BSC), we randomized 35 cancer patients to receive CSF at day 1 (group 1, n = 19) or at day 6 (group 2, n = 16) after transplantation and here we present their hematological reconstitution. BSC collection was performed by apheresis after G-CSF priming for 5 or 6 days (600 micrograms daily subcutaneously). Hematological recovery is comparable between the two groups: a median of 10 (range 7-16) vs 11 (range 9-18) days to reach an ANC > 0.5 x 10(9)/1 in group 1 (G-CSF day 1 after transplant) vs group 2 (G-CSF day 6 after transplant, P = NS). Median time to reach an unsupported platelet count of 25 x 10(9)/1 was 14 days in the two groups (range 8 110 and 10-40 respectively, P = NS); patients received less G-CSF after transplantation in group 2. No difference appeared in terms of transfusion support, number of days of fever of i.v. antibiotic treatment. Patients' hospital stay was the same in the two groups. Our data suggest that delaying G-CSF administration after infusion of mobilized blood cells is not detrimental to hematological recovery, while it lowers the overall cost of the procedure. PMID- 8722351 TI - Related donor marrow transplant for chronic myeloid leukemia: patient characteristics predictive of outcome. AB - Pre-transplant characteristics of 137 consecutive patients (including 103 patients with one or more features suggesting advanced disease) undergoing related donor marrow transplant for chronic myeloid leukemia (CML) were analyzed to determine their association with outcome. Multivariate analysis identified increased recipient age (relative risk (RR) for patients over 30 years of relapse or death 2.37; P = 0.004), and longer interval between diagnosis and transplant (RR 1.20; P = 0.0001) as significant adverse influences on disease-free survival (DFS). The 5-year DFS for patients transplanted within 1 year of diagnosis (?early transplant', n = 71) was significantly higher (51%) than that for patients transplanted beyond 1 year from diagnosis ('delayed transplant', n = 55) (34%; log rank P = 0.02). For early transplant patients, poor prognostic features included myelofibrosis (RR 3.53; P = 0.018), splenomegaly (RR 2.22; P = 0.029) and the use of a female donor (RR 3.16; P = 0.002). The 5-year DFS for patients transplanted within 1 year of diagnosis and without signs of advanced disease was 67%. The presence of increasing numbers of features suggesting acceleration prior to transplant had a cumulative adverse influence of DFS. The risk of relapse (5 year estimate 20%) was also independently and significantly increased in association with a longer interval from diagnosis to transplant (P = 0.012). Early transplant is an important influence on DFS and relapse after related donor transplant therapy for CML, although additional patient characteristics influencing outcome can be identified and may have cumulative adverse effects. PMID- 8722352 TI - Bone marrow 'boosts' following T cell-depleted haploidentical bone marrow transplantation. AB - Nine patients transplanted for non-malignant conditions (eight severe combined immunodeficiency, one aplastic anemia) received lectin-treated T cell-depleted BMTs from haploidentical donors. Each patient subsequently received a second T cell-depleted transplant (?boost') from the same donor, without conditioning, because of a delay in the recovery of T cell immunity associated with evidence of engraftment. The median time to boost after initial BMT was 0.5 years (range 0.2 4.6 years). No conditioning therapy was used prior to the boost (except one patient who received ATG) and no GVHD prophylaxis was used during either the initial or subsequent BMTs. Eight of the nine patients are surviving at a median follow-up of 2.9 years (range 0.3-6.2 years). Following BMT boost, T cell function improved with the lymphocyte proliferative responses to PHA, PWM and alloantigen all increasing at least eight-fold. The mean percentage of CD3+ lymphocytes increased from 20 +/- 7% of total lymphocytes following the first BMT to 66 +/- 7% following the marrow boost (P < 0.001). This increase was generated primarily by an increase in the CD4+ lymphocyte subset which increased from 13 +/ 3% post-transplant to 44 +/- 6% after the BMT boost (P < 0.005), and was reflected in both the CD4+/Leu8+ and CD4+/Leu8- lymphocyte populations. Measurements of B cell immunity (immunoglobulins, isohemagglutinins and B cells) showed no significant effect of the marrow boosts. These results suggest that bone marrow 'boosts' are an effective means for improving T cell immunity in patients who fail to recover adequate immune function after T cell-depleted bone marrow transplantation and may be applicable as immunotherapy following allogeneic BMT undertaken to treat malignancy. PMID- 8722353 TI - Unrelated donor bone marrow transplantation without T cell depletion using a chemotherapy only condition regimen. Low incidence of failed engraftment and severe acute GVHD. AB - Twenty-five patients with hematologic malignancies were treated with busulfan (16 mg/kg) and cyclophosphamide (50 mg/kg x 3 days) as conditioning for bone marrow transplantation using marrow from serologically matched, DR locus genotypically identical unrelated donors. Previous studies of BuCy2 as conditioning for UD-BMT have reported a graft failure rate of up to 21% suggesting it may be insufficiently immunosuppressive in this setting. We elected not to use BuCy4 as it may have a higher incidence of extramedullary toxicity. In addition the patients received GM-CSF (500 mg/m2) from day 0, cyclosporine and short-course methotrexate (15 mg/m2 x 1, then 10 mg/m2 x 3) as GVHD prophylaxis and prophylactic ganciclovir at engraftment if either they or their donor were CMV antibody positive. The median age of the 25 patients was 41 years and the most common diagnosis was CML (76%). Seven patients were considered poor risk and eight males were recipients of marrow from female donors. Sixteen patients survive at a median of 435 days from transplant. The actuarial overall and disease-free survivals at 1 year in this group of older patients were 62 +/- 20% and 57 +/- 20% and 100-day survival was 70%. The engraftment rate was 100%; there have been no instances of secondary graft failure. Fifteen patients (60%) developed grade II-IV GVHD and 12 of 16 (75%) developed some chronic GVHD but only half of these were extensive. The performance status of survivors is good (median of 90); seven of 12 eligible patients are back at work. This study demonstrates that UD-BMT can be successfully performed in very closely HLA matched older patients using a chemotherapy-only protocol and that low rates of severe acute GVHD can be achieved without T cell depletion. PMID- 8722354 TI - High incidence of chronic GVHD after primary allogeneic peripheral blood stem cell transplantation in patients with hematologic malignancies. AB - To assess feasibility and potential advantages of PBSC allograft, we transplanted nine patients (age 20-47 years) with advanced or poor-risk hematologic malignancies. These included eight HLA-identical sibling transplants and one partially matched. Cells were collected from donors by apheresis after rh-G-CSF 10-16 micrograms/kg/day for 4-5 days, and stored at 4 degrees C until infusion. Patients were conditioned with busulfan 16 mg/kg and cyclophosphamide 200 mg/kg, and received GVHD prophylaxis with CSA-MTX. The graft consisted of PBSC alone, with a median of 101.2 (range 28-254.2) x 10(4)/kg CFU-GM, 6.84 (range 4.57-15.9) x 10(6)/kg CD34+ cells and 2.5 (range 1.2-6) x 10(8)/kg CD3+ cells. An ANC > 0.5 x 10(9)/1 occurred on (median) day 13 range 11-17), and a platelet count > 50 x 10(9)/l on (median) day 15 (range 12-29) post graft. One patient died of ARDS on day 13, the others are alive 96-485 (median 245) days from the graft. Two patients have relapsed, one of them with isolated CNS involvement. Acute GVHD (grade I-II) occurred in three patients and severe chronic GVHD in six patients, with no relationship to CSA withdrawal. This unexpected incidence of chronic GVHD might be linked to the high number of CD3+ cells in the graft, contributing to a favourable GVL effect. PMID- 8722355 TI - Cardiopulmonary function and autologous bone marrow transplantation: results and predictive value for respiratory failure and mortality. The University of Nebraska Medical Center Bone Marrow Transplantation Pulmonary Study Group. AB - Cardiopulmonary complications are a major cause of morbidity and mortality in patients undergoing high-dose therapy with stem cell transplant support. Since exercise tolerance testing (ETT) assesses the cardiopulmonary reserve of an individual, we hypothesized that ETT performed prior to transplant would predict respiratory failure and mortality and would be a superior predictor over resting cardiopulmonary function tests. We performed a retrospective study of 191 lymphoma patients who underwent ETT prior to transplant between 1 June 1990 and 31 December 1992 and compared the results of ETT with resting pulmonary function tests (PFT) and resting cardiac ejection fraction (EF). ETT revealed that cardiac, pulmonary and combined cardiopulmonary limitation were observed in 31, 20 and 16% of the patients, respectively, with a gas diffusion-type limitation being the most common exercise limitation. Resting PFT were abnormal in 58% of patients with a diffusion defect being the most common abnormality. Low EF was observed in 6.8% of patients. Twelve patients eventually required mechanical ventilation post-transplant with only the resting diffusion PFT predictive of this complication. There were five early deaths that were attributable to respiratory failure and neither resting nor ETT studies were predictive of these deaths. ETT and EF performed prior to transplant in lymphoma patients undergoing autologous transplant do not predict for either respiratory failure or short-term mortality. Our findings may be due to the rather low incidence of respiratory failure (6.3%) and low early mortality from cardiopulmonary complications (2.6%) seen in our patient population. PMID- 8722356 TI - Autologous bone marrow transplantation is feasible in patients with a prior history of invasive pulmonary aspergillosis. AB - We report on seven adult leukemic patients who were autografted in spite of a prior history of invasive pulmonary aspergillosis (IPA). Their median age was 41 years (range: 19-61); six patients were male and one female. All seven had acute myeloblastic leukemia (AML) and underwent an autologous marrow transplantation (ABMT) with a marrow purged in vitro by mafosfamide. IPA was suspected prior to ABMT on clinical and radiological features. CT scan confirmed nodular infiltrates and cavitations in six cases. Microbiological documentation consisted of: identification of the fungus from bronchoalveolar lavage: one case, positive antigenemia: one case, positive antibodies: two cases. Prior ABMT patients received amphotericin B for a median total dose of 1915 mg (range: 970-3300). No patient underwent surgery. The median time from diagnosis of IPA to ABMT was 7.3 months (range: 3-10). During ABMT all patients received prophylactic amphotericin B and itraconazole. No patient died from toxicity and no IPA reactivation was observed in any patients. Post-graft, itraconazole was kept on for a median of 3 months (range: 3-5). This study demonstrates that IPA occurring during the management of AML patients is not necessarily a contraindication to subsequent ABMT. PMID- 8722357 TI - Presence of human polyomavirus DNA in the peripheral circulation of bone marrow transplant patients with and without hemorrhagic cystitis. AB - In BMT patients, shedding of BK virus (BKV) in the urine has been strongly but not absolutely correlated to hemorrhagic cystitis (HC). The possible presence of human polyomaviruses in peripheral blood leukocytes (PBLs), plasma, serum and urine in BMT patients and an association with HC was investigated by a nested PCR assay. Samples from allogeneic BMT patients with and without HC as well as from autologous BMT patients were analyzed. Human polyomaviruses were detected in urine and blood samples of both allogeneic and autologous BMT patients with and without HC. An association between the presence of a specific human polyomavirus in blood and HC was thus not observed. PMID- 8722358 TI - Activation of coagulation and deep vein thrombosis after bone marrow harvesting and insertion of a Hickman-catheter in ABMT patients with malignant lymphoma. AB - Evidence of activation of coagulation was sought in serial plasma samples from 25 ABMT candidates with malignant lymphoma admitted for bone marrow harvesting: 10 females and 15 males, median age 41 years (range 27-58 years). Nineteen patients had non-Hodgkin's lymphoma (NHL) and six had Hodgkin's disease. Of those with NHL, 14 had high-grade and five low- grade disease. The plasma levels of markers of activation (prothrombin fragment 1 + 2, thrombin-antithrombin complexes, fibrinopeptide A and fibrinmonomers) increased significantly (P < 0.001) in association with harvesting. Except for fibrinopeptide A, the indicators of activation were still significantly elevated 24 h after marrow aspiration. Beta thromboglobulin, a marker of the platelet release reaction, also increased significantly (P < 0.01). Four out of nine patients in whom a long-term central venous catheter was inserted just after marrow aspiration, developed catheter related deep vein thrombosis, verified venographically, shortly after harvesting. These results suggest that patient with malignant lymphoma undergoing marrow harvesting develop a hypercoagulable state, and that insertion of a central intravenous catheter immediately after marrow harvesting should be avoided to prevent the development of symptomatic deep vein thrombosis. PMID- 8722359 TI - Use of a semi-quantitative PCR for cytomegalovirus DNA as a basis for pre-emptive antiviral therapy in allogeneic bone marrow transplant patients. AB - The aim of this study was to evaluate the efficacy of pre-emptive antiviral therapy, based on a semi-quantitative nested PCR for cytomegalovirus (CMV) DNA in leukocytes, for the prevention of CMV disease after allogeneic BMT. Fifty-eight patients were prospectively followed with PCR for CMV DNA and antiviral therapy with ganciclovir was initiated after two consecutive positive tests. The levels of CMV DNA were determined by serial dilutions of the positive samples. The probability of detection of CMV DNA was 48.3% and the probability of CMV disease 6% at 100 days after BMT. Patients with CMV disease had higher CMV DNA levels compared with patients without CMV disease (P = 0.001). In comparison to 58 matched historical controls detection of CMV DNA was 5 days earlier (NS) and antiviral therapy could be initiated 10 days earlier in patients followed by PCR (P = 0.05). Pre-emptive antiviral therapy was given to 28 patients in a total of 36 courses. Patients became negative in PCR after 28 of 36 courses (77%). We conclude that PCR for CMV DNA can be used for early detection of CMV infection and as the basis of initiation of pre-emptive antiviral therapy in BMT patients. PMID- 8722360 TI - Ganciclovir prophylaxis for cytomegalovirus infection in pediatric allogeneic bone marrow transplant recipients. AB - Twenty-nine pediatric allogeneic bone marrow transplant (BMT) recipients, ages 2 17 years, were followed prospectively for cytomegalovirus (CMV) infection. Patients at risk received ganciclovir (GCV) prophylactically at a dose of 5 mg/kg/day i.v., 3 to 5 days per week, until day 100. Surveillance blood and urines were obtained weekly. Twelve patients developed DMV infection: one patient died with CMV interstitial pneumonitis on day 19 post-transplant prior to initiating GCV prophylaxis; 10 patients developed CMV viremia (n = 9) or viruria (n = 1) between day 30 and day 95 (median day 50) while receiving GCV prophylaxis; and one patient developed asymptomatic CMV viruria on day 130, 1 month after completing GCV prophylaxis. Patients with breakthrough infections on prophylaxis were treated with intensified GCV and i.v. immunoglobulin. No patient developed visceral involvement, although five patients had recurrent viremia. Six of the seven long-term survivors continued to excrete CMV in the urine intermittently for 6 to 28 months post-transplant. GCV was well tolerated with transient, mild neutropenia in five patients and thrombocytopenia in four patients. No extramedullary toxicity was encountered. GCV prophylaxis at a dose of 15-25 mg/kg/week is not adequate to prevent CMV reactivation in children receiving marrow transplants from unrelated donors and/or T cell-depleted grafts. PMID- 8722361 TI - Sexual dysfunction prior to high-dose chemotherapy and bone marrow transplantation. AB - A number of studies have suggested that high-dose chemotherapy and bone marrow transplantation may be associated with a variety of abnormalities of psychological function including sexual dysfunction. However, few studies have prospectively evaluated the association between sexual dysfunction and BMT. In particular, there are very little baseline data about sexual function immediately before transplant. The sexual function of 30 patients was assessed immediately prior to high-dose chemotherapy and bone marrow transplantation using the Derogatis Interview for Sexual Functioning (DISF) for males and females. More than 80% of patients had hematological malignancies; more than half were allograft recipients. Forty-seven percent of patients were found to have global sexual dysfunction and 60% had abnormalities of at least one parameter of sexual function. Forty-seven percent were dissatisfied with their sex life. Sexual dysfunction was associated with ejaculatory problems (P < 0.02) and erectile problems (P = 0.06) but not with amenorrhea. There was an association between cancer-related psychological problems and sexual dysfunction. A control group of inpatients with cancer had a similar incidence of sexual dysfunction (53% vs 47%, P = NS) suggesting that the tumor and its therapy were the major reasons for the sexual problems and not the prospect of transplant per se. This study emphasizes the need for baseline (pre-BMT) studies of quality of life and psychological function in BMT patients. We conclude that sexual dysfunction is a common finding prior to BMT; whether intervention can reduce this problem requires further study. PMID- 8722363 TI - Absence of cytomegalovirus and Epstein-Barr virus expression in labial salivary glands of patients with chronic graft-versus-host disease. AB - We investigated in 15 consecutive patients a possible correlation between expression of CMV or EBV in labial salivary gland (LSG) biopsies performed 100 days after allogeneic BMT and subsequent development of chronic GVHD. Three techniques were performed for the detection of each virus: immunohistochemistry, in situ hybridization and PCR. Eleven patients developed chronic GVHD. Histologic examination detected a moderate lymphoid infiltrate (grade 1 according to Sale's score) in LSG biopsy in only one patient. CMV genes or proteins could not be detected in any patients. Likewise, EBV genome or proteins were not detected by in situ hybridization and immunohistochemistry. However, in three of the 15 patients, EBV DNA was detected by PCR in LSG biopsies. Only one of these three patients developed chronic GVHD. Therefore, at the present time, the presence of a lymphoid infiltrate on lip biopsies performed at day 100 post-BMT does not appear to be sensitive enough for the diagnosis or the prediction of the subsequent development of chronic GVHD. Moreover, the absence of EBV and CMV expression in a day-100 LSG biopsy does not preclude the development of chronic GVHD. PMID- 8722362 TI - Hepatitis C virus (HCV) infection in bone marrow transplant patients after transfusions from anti-HCV-positive blood donors. AB - In March 1992, 12 bone marrow transplant patients at the Fred Hutchinson Cancer Research Center received blood components from donors who were anti-HCV nonreactive by first generation ELISA but whose serum later tested anti-HCV reactive to a second generation ELISA. All these blood components were further tested for anti-HCV using a second-generation RIBA and for HCV RNA by polymerase chain reaction. Recipient sera were tested for HCV RNA prior to and following blood component infusion. Blood components from four donors were positive for HCV RNA. All recipients of HCV RNA-positive blood components became viremic on the first day tested post-infusion. In addition, two recipients of HCV RNA-negative blood components tested HCV RNA-positive both pre- and post-infusion. Viremia persisted up to the time of death or day 100 in five of the six patients who were HCV RNA-positive post-transplant. No HCV RNA-positive recipient developed symptomatic acute hepatitis, and only two had aminotransferase elevations consistent with chronic hepatitis. We conclude that HCV RNA-positivity in blood components accurately predicts transmission of virus. Infection with HCV did not adversely affect short-term patient outcome following bone marrow transplantation. PMID- 8722364 TI - Acquired ciliary abnormalities of nasal mucosa in marrow recipients. AB - Respiratory symptoms are frequent after bone marrow transplantation (BMT). Most studies focus on lesions of the lower respiratory tract. However, sinusitis is also common in this setting, especially after allogeneic BMT. The nasal respiratory epithelium is the first line of airway defense and is very similar to the bronchial epithelium, especially in terms of ciliary beat frequency and ultrastructural pattern of ciliated cells. We have prospectively studied the nasal respiratory epithelium of 20 marrow recipients (four autologous, 16 allogeneic) with or without sinusitis, by brushing and biopsy of the median turbinate between 2.5 and 148 months after transplant. Samples were studied for ciliary beat frequency, cytology, ultrastructural pattern and HLA-DR expression. We found that 17 of our 20 patients had abnormalities of their nasal epithelium, mainly consisting of either squamous metaplasia or heterogeneous axonemal defects of peripheral and central microtubules. No relationship between these findings and the presence of acute or chronic sinus infection, previous irradiation, graft versus-host disease or immunosuppressive therapy could be demonstrated in this preliminary study. These abnormalities probably have multiple causes. Prospective studies are needed to determine the respective roles of treatments, infections and immune disorders associated with BMT in these abnormalities, and to know their natural evolution over time and their impact on the occurrence of upper or lower respiratory tract infections. PMID- 8722365 TI - Antibodies to interferon-alpha: a novel type of autoantibody occurring after allogeneic bone marrow transplantation. AB - Antibodies to IFN-alpha have been recognized as a novel type of autoantibody developing after allogeneic BMT. Ninety-six patients undergoing BMT for various hematologic disorders were followed for the presence of spontaneous IFN-alpha antibodies until 12 years after transplantation. Seven of them (7.3%) developed IFN-alpha antibodies occurred late after BMT (> or = 15 months), rose to very high titers in some patients, and persisted for years despite combined immunosuppressive treatment. They were oligo- or polyclonal in nature, predominantly IgG with a broad IgG subclass distribution, and neutralized the antiviral and antiproliferative activity of various natural and recombinant IFN alpha types including the patients' endogenous IFN-alpha in vitro. All antibody positive recipients suffered from chronic GVHD (n = 5) or chronic viral hepatitis (n = 2), but the only significant association was with prior severe aplastic anemia (3/9, 33%; P = 0.022). There was no discernible HLA association of IFN antibody development. Although the clinical relevance of the IFN-alpha antibodies is uncertain they may interfere with cellular defence mechanisms and immune regulation after BMT. PMID- 8722366 TI - Monitoring and prognostic evaluation of sex-mismatched bone marrow transplantation by competitive PCR on Y chromosome sequences. AB - In one case out of four, allogeneic BMT concerns a male recipient and a female donor. The monitoring of sex-matched BMT can be carried out by PCR amplification on Y-specific chromosome sequences (YCS), whatever the hematological disease. Twelve patients with sex-mismatched non-T-depleted BMT were first studied through a qualitative PCR, which gave semi-quantitative results. When the qualitative PCR revealed YCS, a competitive amplification was performed in order to estimate the YCS amount in the patient blood sample. For the purpose of the study, we classified the patients in two categories according to the results obtained 9 months after BMT. For 10 patients, we did not detect any YCS amplification after this time. These patients were in complete cytogenetic and clinical remission. For the remaining two patients, we always found male DNA in their blood samples. These patients were in cytogenetic remission but relapsed and died 21 and 25 months after BMT. Our results suggest that the persistence of male cells in peripheral blood, even at the low rate of 1% or 0.1%, 1 year after sex-mismatched BMT, is a bad prognosis. PMID- 8722367 TI - Response to measles, mumps and rubella vaccine in paediatric bone marrow transplant recipients. AB - Bone marrow transplant (BMT) recipients are routinely reimmmunized with the childhood vaccine series after transplantation excluding the live attenuated vaccines. In this study, the clinical and serologic responses to measles, mumps and rubella (MMR) vaccine in children after BMT was assessed. Twenty-two BMT recipients were vaccinated with MMR II (MSD). All were at least 2 years post-BMT and without GVHD. Their underlying conditions were leukemia (11), aplastic or Fanconi's anemia (7), thalassemia (3) and metabolic disease (1). All were allogeneic transplants with matched related donors. The mean age at transplantation was 6.9 years. There were no reported adverse effects of the vaccination. Antibody status for MMR was determined using commercial assays (IFA and ELISA) on paired specimens. The mean interval between transplantation and vaccination was 48 months. Pre-vaccination, no BMT recipient was sero-positive for all three, but 23% were positive for measles, 31% for mumps and 14% for rubella. Post-vaccination, 68% of BMT recipients were sero-positive for all three, with 77% for measles, 87% for mumps and 91% for rubella. Therefore, MMR vaccination at 2 years or later after BMT in paediatric recipients without GVHD was safe and significantly increased the proportion sero-positive for MMR. PMID- 8722369 TI - Polymerase chain reaction is highly predictive of relapse in patients following T cell-depleted allogeneic bone marrow transplantation for chronic myeloid leukemia. AB - In chronic myeloid leukemia (CML) the polymerase chain reaction (PCR) can be used to detect minimal residual disease after bone marrow transplantation (BMT). Previous studies have shown that PCR positivity is common following BMT. However, the clinical significance of this finding for any given individual who is PCR positive remains unclear, as many of these patients remain long-term disease-free survivors after allogeneic BMT. In the present study, we used PCR to detect BCR ABL mRNA in 144 blood or marrow samples from 36 patients who received a T cell depleted BMT for CML in first chronic phase. Six patients had no evidence of PCR detectable residual disease at any time following transplant. The other 30 patients had at least one positive PCR result post-BMT. Once PCR positivity was found, it was usually sustained, with only four patients having a subsequent PCR negative assay. No patient who had two consecutive PCR-positive assays had a return to PCR negativity. None of the six patients with exclusively PCR-negative assays have developed either cytogenetic or hematologic relapse at a median follow-up of 42 months. Of the 30 patients with at least one PCR-positive assay post-BMT, 28 were PCR positive at last follow-up, and 22 have progressed to cytogenetic or hematologic relapse. If the PCR-positive assay occurred within 24 months of the transplant then the estimated probability of progression to cytogenetic or hematologic relapse was 65% at 24 months. Twenty of the 26 patients who were studied early (< or = 6 months) after BMT had at least one positive PCR assay. Fifteen of the 20 patients who were PCR positive < or = 6 months following transplant have progressed to either cytogenetic or hematologic relapse resulting in an estimated probability of relapse of 84% at 24 months. These results indicate that following T cell-depleted BMT for CML in first chronic phase, PCR is highly predictive of relapse and may identify a cohort of patients in need of therapeutic intervention before the onset of overt clinical relapse. PMID- 8722368 TI - Central nervous system relapse after bone marrow transplantation for acute leukemia in first remission. AB - Four hundred and eighty-seven patients undergoing allogeneic or autologous BMT for acute leukemia in first remission received no prophylactic intrathecal chemotherapy after BMT. The conditioning regimen included total body irradiation in 433 (89%). Patients with acute lymphoblastic leukemia received cranial irradiation if they had no central nervous system (CNS) disease and all patients with CNS disease received craniospinal irradiation. Eleven of 311 patients examined had CNS disease at presentation, but none had active CNS disease at the time of BMT. Lumbar punctures were performed in 93 patients 1-2229 days (median 98) after BMT because of suspected CNS infection, hemorrhage or relapse (n = 65), or after systemic relapse (n = 28). CNS disease was seen in seven patients at 217 1209 days (median 340), none of whom had CNS disease pre-transplant. Systemic relapse had preceded CNS relapse by 4-207 (median 128) days in all seven and no isolated CNS relapse was seen. The actuarial risk of CNS and systemic relapse at 5 years was 2.9 and 35.9%, respectively. We conclude that CNS relapse is uncommon after BMT for acute leukemia in first remission, and that isolated CNS relapse is likely to be extremely uncommon. Therefore, routine prophylactic intrathecal chemotherapy is not warranted after BMT for acute leukemia in first remission. PMID- 8722370 TI - Tacrolimus (FK506) and methotrexate regimens to prevent graft-versus-host disease after unrelated dog leukocyte antigen (DLA) nonidentical marrow transplantation. AB - We previously reported an synergism between methotrexate and tacrolimus (FK506) in preventing graft-versus-host disease (GVHD) in dogs given DLA-nonidentical unrelated marrow grafts after 9.2 Gy of total body irradiation (TBI). Methotrexate was given at 0.4 mg/kg i.v. on days 1, 3, 6 and 11 and FK506 at 0.15 mg/kg/day i.m. on days 0-8 and 0.5 mg/kg/day orally on days 9-90. Half of the dogs became long-term survivors. A major toxicity was gastrointestinal, and 25% of dogs died with intussusception. The current study addresses the problem of intussusception by making changes in drug doses used. In one group of dogs, FK506 was reduced to 0.075 mg/kg i.m. on days 1-8, while methotrexate was administered per original schedule. In a second group, methotrexate was reduced to a single dose on day 7, while FK506 was either administered per the original or reduced dose schedule. None of the 17 current dogs developed intussusception, however, all but two dogs died with GVHD (n = 12) or graft failure (n = 3). Only two dogs survived after transient GVHD. Results show that there is little room for maneuvering FK506 or methotrexate doses, and hopes of reducing gastrointestinal toxicity by dose modifications while retaining the ability to prevent GVHD were not fulfilled. PMID- 8722371 TI - Laparoscopic liver biopsy to evaluate hepatic dysfunction in patients with hematologic malignancies: a useful tool to effect changes in management. AB - Hepatic dysfunction is common in patients who receive intensive chemotherapy and it is important to determine the etiology in order to institute appropriate therapy. The role of laparoscopic liver biopsy in patients with neutropenia, thrombocytopenia, or both was evaluated as a mean of making treatment decisions and as a determinant of clinical outcome. Laparoscopic liver biopsy was performed in 29 subjects who were receiving intensive cytotoxic therapy with or without bone marrow transplantation. One to three direct-vision laparoscopic liver biopsies were performed in each patient using a Tru-cut needle during general anesthesia. Platelet concentrate transfusions were usually given before, during, and immediately after biopsy. Bleeding was controlled with spatula electrocautery. Thirty-two biopsies were obtained in 29 patients. At the time of liver biopsy, white blood cell and platelet counts ranged from 0 to 14,300/microliters (median: 2500/microliters), and 1000 to 47,000/microliters (median: 20,000/microliters), respectively. Bleeding at the liver biopsy site was readily controlled during the procedure without clinical evidence of significant bleeding; no procedure-related complications were noted and no patients required re-exploration. All biopsies were informative and the lesions observed in 32 biopsies revealed graft-versus-host disease (n = 5), hepatic candidiasis (n =1), hepatic veno-occlusive disease (n = 3), cholestasis (n = 19), hemosiderosis (n = 26), toxic injury (n = 8), hepatic steatosis (n = 4), granuloma (n = 1), viral infection (n =1), and malignancy (n = 1). Laparoscopic liver biopsy has been proven to be an effective means of assessing the cause of liver dysfunction in patients who were thrombocytopenic and immunosuppressed. The diagnosis obtained at laparoscopic liver biopsy altered therapy in nine of 29 (31%) patients. PMID- 8722372 TI - Tunneled, double lumen Broviac catheters are useful, efficient and safe in children undergoing peripheral blood progenitor cell harvesting and transplantation. AB - We prospectively evaluated performance, efficiency, safety and compliance of large-bore central venous catheters (Cook TPN; Cook (Switzerland) AG) introduced via saphenous veins in 32 children and infants elected to undergo peripheral blood progenitor cell (PBPC) harvesting and transplantation (PBPCT). With these catheters a flow rate (25-65 ml/min) adequate for leukapheresis was achieved in all patients. There were no important catheter-related complications during harvest. The total duration of catheter placement was 4569 days (median, 139 days; range, 8-268 days). During this period which included conditioning and PBPCT, we observed five mechanical complications and 12 septic episodes not necessarily catheter-related (0.11 and 0.26 events per 100 catheter days, respectively). All infections resolved after systemic antibiotic treatment. There was no exit or tunnel area infection, and no catheter had to be removed due to infection. Two catheters were replaced because of displacement. Tunneled double lumen Broviac catheters introduced via saphenous veins were not only efficient and safe, they were also well accepted by children and young adults undergoing PBPC harvesting and transplantation. PMID- 8722373 TI - Kaposi's sarcoma after autologous bone marrow transplantation for multiple myeloma. AB - We report a case of Kaposi's sarcoma in a patient who underwent autologous bone marrow transplantation (ABMT) for multiple myeloma. Four months after ABMT he presented with numerous asymptomatic, dark blue and purplish macules and nodules on the trunk and lower extremities. Biopsy revealed the typical histologic pattern of Kaposi's sarcoma. The patient died due to disseminated Kaposi's sarcoma while in complete remission of his hematologic malignancy. PMID- 8722376 TI - Immune and inflammatory responses to Helicobacter pylori infection. AB - The gastroduodenal response to chronic Helicobacter pylori infection is characterized by the infiltration of plasma cells, lymphocytes, neutrophils and monocytes into the mucosa. Eradication studies have shown that this inflammatory response represents a specific reaction to the presence of H. pylori. As well as stimulating specific local T and B cell responses and a systemic antibody response, H. pylori infection also induces a local pro-inflammatory cytokine response. Interleukin-8 (IL-8), which is expressed and secreted by gastric epithelial cells, may be an important host mediator inducing neutrophil migration and activation. IL-8 mRNA and protein secretion in gastric epithelial cell lines can be up-regulated by the cytokines tumour necrosis factor-alpha and IL-1 and also by type I strains of H. pylori (expressing the vacuolating toxin and cytotoxin-associated protein, CagA). The gastric epithelium thus plays an active role in mucosal defence. Neutrophil activation and the production of reactive oxygen metabolites will be induced directly by bacterial factors and indirectly via host-derived cytokines, products of complement activation and bioactive lipids. Strain variation in the induction of both IL-8 from epithelial cells and the oxidative burst in neutrophils may be an important factor determining the extent of mucosal injury. There is now increasing evidence from both in vivo and in vitro studies that type I strains induce an enhanced inflammatory response and mucosal damage. An understanding of the bacterial mediators of mucosal inflammation is important in elucidating the role of chronic H. pylori infection in the pathogenesis of gastroduodenal disease. PMID- 8722374 TI - Peripheral blood progenitor cell (PBPC) transplantation with a single apheresis in various malignancies. PMID- 8722377 TI - Prevention of Helicobacter pylori infection. AB - The only successful strategy for large-scale eradication of an infectious disease from whole populations has been through immunization. In societies fortunate enough to have a healthcare infrastructure that allows mass vaccination, diseases such as poliomyelitis, whooping cough and diphtheria have been virtually eliminated. But what about infection with Helicobacter pylori, which has now been proved to be a major worldwide pathogen? Infection by H. pylori is lifelong, despite a vigorous immune response. How, then, could immunization be feasible? Recent investigations using a mouse model of H. pylori infection have shown that protective immunity can indeed be induced with an oral vaccine. Furthermore, protection was lifelong in the animal model. Also, more importantly, the vaccine actually cured existing infection, raising the exciting possibility of therapeutic immunization. Stimulated by the medical need, the race is now on to produce the first vaccine. However, many critical questions remain to be answered before the first patients are immunized with the complete vaccine. For example, what is the best antigen to target? Urease is currently the favourite antigen, but others will be tested. What about the adjuvant? LT, the heat labile enterotoxin of diarrhoeaogenic strains of Escherichia coli, is the most likely candidate, but we need to examine alternatives. Is there any risk of immunopotentiation? The answer is probably no, but this has to be proved. A vaccine against H. pylori will be developed, and infection with H. pylori will be prevented--the question remains 'how long will it take'? PMID- 8722378 TI - Pathological consequences of Helicobacter pylori infection. PMID- 8722379 TI - Pathogenic properties of Helicobacter pylori. AB - Helicobacter pylori possesses factors that allow it to colonize the gastrointestinal mucosa and persist at that site. Here it produces adverse pathological changes, and thereby causes disease. Colonization factors: animal models have shown that motility and the production of urease are essential for colonization by H. pylori. The ability of an organism to adhere to host structures is often considered pivotal in colonization. A number of adhesins associated with H. pylori have been described, which may imply that adherence is a multistep process and that different adhesins mediate adherence to different sites in the gastric tissue. Persistence factors: H. pylori lipopolysaccharide (LPS) possess low immunological activity, thereby minimizing the local inflammatory response and contributing to the persistence of the infection. There is also evidence that the LPS affects the qualitative nature of gastric mucin and stimulates pepsinogen secretion. Whether survival during exposure to antimicrobial agents is aided by the development of coccoid forms with intact membranes and polyphosphate energy reserves is not yet known. Putative disease inducing factors: these include the vacuolating cytotoxin that is capable of inducing gastric ulceration in mice, ammonia products that induce vacuolation, and phospholipases that may affect the hydrophobicity of the mucosa. Mimicry of Lewis blood group antigens on the surface of H. pylori may also contribute to pathogenesis. Characteristics of certain strains, such as the expression of a cytotoxin-associated gene (cagA) and the ability to induce rapid chemiluminescence in neutrophils, are associated with the induction of peptic ulceration. PMID- 8722380 TI - Helicobacter pylori and disturbance of gastric function associated with duodenal ulcer disease and gastric cancer. AB - Helicobacter pylori is now recognized as the major acquired factor in the pathogenesis of duodenal ulcer disease (DU). There is also an association between H. pylori infection and the subsequent development of gastric cancer. The mechanisms by which such infection predisposes the host to these diseases are incompletely understood, but disorders induced by the bacterium in gastric function play a pivotal role. In most patients, H. pylori infection stimulates acid secretion, leading to a predisposition to DU development. However, in some patients, the infection is associated with a significant decrease in acid secretion, a predisposition to gastric cancer. These divergent effects of H. pylori on gastric acid secretion explain the early conflicting reports on changes in acid secretion associated with the infection. The reason why H. pylori infection produces divergent effects on gastric acid secretion is unclear, but may be related to differences in bacterial strains or genetic, dietary or other environmental factors. PMID- 8722381 TI - Helicobacter pylori infection and dyspepsia. AB - It has proved remarkably difficult to confirm or refute an association between Helicobacter pylori-induced gastritis and non-ulcer dyspepsia for several reasons. Dyspepsia has not been defined adequately and current definitions include a variety of symptoms that probably reflect a number of underlying pathophysiological processes. Dyspepsia is a symptom complex, rather than a specific disease entity, and cannot be easily identified. There are probably many causes of dyspepsia, and if H. pylori does cause symptoms, it may produce different symptoms in different people. However, little is known about variations in host susceptibility and H. pylori virulence, or their potential effect on symptomatology. In addition, the mechanisms responsible for dyspeptic symptoms are unknown, regardless of whether or not there is concomitant H. pylori-induced inflammation. A possible aetiological role for H. pylori in dyspepsia has been sought in three major types of study. Epidemiology studies have suggested that H. pylori is not a major cause of non-ulcer dyspepsia. However, both dyspepsia and H. pylori are common, and the studies cannot account for many of the variables detailed above. Pathophysiological studies have commonly reported a variety of motor and sensory 'abnormalities' in association with H. pylori, but none has been confirmed or reproduced. In eradication studies, there is continuing debate as to whether dyspeptic symptoms diminish with treatment. This is partly because of the high placebo-response rate and partly because many treatment regimens have not cured the infection. Suppression of bacterial growth may not affect symptoms significantly, if they are due to mucosal inflammation, and symptom resolution may take many weeks or months following cure of H. pylori and the associated gastritis. Recent findings have shown that symptom reduction is more evident 1 year after eradication of H. pylori than after 4 weeks. There is a continuing and urgent need for well-designed studies to assess the long-term effect of H. pylori and its cure on both gastrointestinal function and dyspeptic symptoms. PMID- 8722382 TI - Helicobacter pylori and gastric cancer. AB - The International Agency for Research on Cancer, sponsored by the World Health Organization, has recently categorized Helicobacter pylori infection as a class I carcinogen, based on evidence that this infection increases the risk of gastric cancer. The classification was intentionally qualitative in nature and not associated with any public health recommendations. In addition, no specific causal mechanism was proposed to explain the relationship between H. pylori and gastric cancer. In this paper, the magnitude of the risk, implications of the relationship for the prevention of gastric cancer and nature of the causal mechanisms are considered. Relative risk of gastric cancer may be substantial; even with conservative assumptions, the proportion of new cases of gastric cancer worldwide attributable to H. pylori infection is approximately one third of a million annually. This figure is likely to increase with changes in the age structure of the population, and the eradication of H. pylori as a means of prevention of gastric cancer should be considered. A strategy of screening populations in middle age and treating those infected could be relatively inexpensive to administer, but the efficacy is totally unknown and requires evaluation in a randomized controlled trial. Studies designed to address this issue in the general population would need to be large and long-term if gastric cancer is used as an end-point. With respect to carcinogenic mechanisms, a number of constitutive properties of H. pylori may be of relevance to cancer without being specifically carcinogenic. Thus ammonia, which is produced in abundance as a result of urease activity, may promote cell division. Other relevant properties result from the immune response of the host to the bacterium. For example, the excessive production of reactive oxygen metabolites can lead to extensive DNA damage and molecular mutations. PMID- 8722383 TI - Diagnosis of Helicobacter pylori infection. PMID- 8722384 TI - Advantages and disadvantages of current diagnostic tests for the detection of Helicobacter pylori. AB - Current tests used to detect Helicobacter pylori are either invasive (histological detection, culture, the polymerase chain reaction (PCR), smear examination) or non-invasive (serology, 13C-urea breath test). These tests vary in their sensitivity and specificity, and the choice of test will depend on the situation, for example, whether the test is to detect infection or the success of eradication treatment. The accuracy of histological tests depends, to a large degree, on the expertise of the pathologist, while the accuracy of culture can depend on the conditions in which the specimen is transported and processed. When performed under optimal conditions, both techniques give very good results. The PCR test has similar sensitivity and specificity to histological and culture tests but a strict protocol must be followed to avoid contamination with H. pylori DNA. The rapid urease test (with a reading taken 1 hour later) is suitable for diagnosis before treatment but its sensitivity decreases after treatment. Smear examination has limited sensitivity. The urea breath test and serology (specific IgG detected by enzyme-linked immunosorbent assay with purified antigens) have sensitivities close to those using the best of the biopsy methods. Other points to consider when selecting a test are its availability, the rapidity of the results (which can range from a few minutes to 2 weeks), possibilities for retrospective analysis, quantification and the detection of pathogenic properties, the globality of certain tests that present an overall picture of the stomach, thus avoiding errors in sampling, and the cost of the test. Important added value can be gained from certain tests: histology allows evaluation of the status of the mucosa, culture allows strain typing and tests for antibiotic susceptibility, and the breath test can confirm successful eradication without endoscopy. When the diagnostic tests are performed correctly, most of them are highly accurate. PMID- 8722385 TI - Diagnosis of Helicobacter pylori infection. When to use which test and why. AB - Tests for the detection of Helicobacter pylori differ in many respects. The choice of test depends on the clinical situation. In symptomatic patients, endoscopy is an important tool for diagnosing peptic ulcer disease or other gastroesophageal lesions. A biopsy-based detection system for H. pylori, such as the rapid urease test, microscopy or culture of the organisms, is therefore appropriate. The diagnostic sensitivity will increase if more than one of these is performed. In asymptomatic patients, a non-invasive test should be used, and serology with a titre is suitable in this situation. If the results prove positive, confirmation of H. pylori infection is recommended using a urea breath test. Patients who have previously received H. pylori eradication therapy and present with a recurrence of symptoms should receive further eradication therapy if they are still H. pylori-positive, even if no endoscopic abnormality is found. A non-invasive test should be used and the urea breath test is the best option in these individuals, as serology is not suitable within 6-12 months of eradication therapy. If the breath test is unavailable, a biopsy-based test should be used. The outcome of H. pylori eradication therapy should be assessed using a non invasive test. The urea breath test is appropriate, as serology is not suitable post-treatment. The breath test should be performed 4-8 weeks after the end of treatment to avoid false-negative results caused by temporary suppression of the infection. If the breath test is unavailable, a serology with a titre or a biopsy based test must be used, at least 6-12 months after the end of treatment. PMID- 8722386 TI - Management of Helicobacter pylori-positive peptic ulcer disease. PMID- 8722387 TI - Current indications for Helicobacter pylori eradication therapy. AB - Two questions need to be answered before the eradication of Helicobacter pylori infection can be discussed. Firstly, can the infection be cured, and if so, is the cure permanent or transient? Secondly, what is the long-term clinical outcome of H. pylori infection? Current indications for the eradication of H. pylori infection can be divided into three categories: well-established, equivocal and experimental. Peptic ulcer disease in patients positive for H. pylori is a well established indication for eradication of the organism. H. pylori-positive dyspepsia and the need for prolonged treatment with non-steroidal anti inflammatory drugs are equivocal indications. MALT (mucosa-associated lymphoid tissue)-lymphoma, hypertrophic/hyperplastic gastritis, atrophic gastritis with or without premalignant mucosal alterations, and the need for prolonged powerful acid-suppressive therapy, in patients positive for H. pylori are all experimental indications. The decision to recommend H. pylori eradication therapy should not be taken lightly. Despite the risks associated with antimicrobial therapy, however, the benefits from cure of the infection are such that antimicrobial therapy is being increasingly favoured, and the reasoning behind withholding eradication therapy is becoming more difficult to understand in the light of current knowledge. PMID- 8722388 TI - Review of Helicobacter pylori eradication regimens. AB - A large number of different treatment regimens aimed at eradicating Helicobacter pylori have been evaluated and reported. A meta-analysis of the pooled results is reviewed here. The treatment strategies have been grouped into 13 treatment classes. The two dual combination therapies reviewed, omeprazole with amoxycillin or clarithromycin, have been analysed with regard to dose and dosing. In all other groups, the total dose of the drugs, number of doses, duration of therapy, and formulation are not taken into account. RESULTS: A sensitivity test comparing the effect of different study designs on eradication rates showed that study quality has no conclusive impact on success rates. The two dual therapies, together with the H2-receptor antagonist triple therapies, are less effective than triple therapies with omeprazole. Bismuth-based triple therapies have a mean overall eradication rate of 79%, but are limited by frequent and significant side effects causing poor drug compliance. Data on lansoprazole triple therapies are almost non-existent, while data on omeprazole triple therapies are extensive and consistently show eradication rates of close to or above 90%. PMID- 8722389 TI - Eradication of Helicobacter pylori: omeprazole in combination with antibiotics. AB - Until recently, the mainstay of treatment for Helicobacter pylori infection was either dual therapy, using omeprazole with amoxycillin or clarithromycin, or traditional triple therapy comprising bismuth and two antibiotics. Success with these treatment strategies has, however, varied widely between centres. Furthermore, the side-effects reported for bismuth triple therapy and the 2-week treatment period recommended have limited its popularity. These drawbacks have thus stimulated research aimed at identifying better drug combinations, with a simpler dosage for a shorter period, fewer side-effects, and greater and more consistent efficacy. A number of studies have now been undertaken using an acid inhibitor in combination with two antibiotics. Omeprazole, a highly effective acid pump inhibitor, has been investigated most extensively in this context, and is markedly effective in eradicating H. pylori when combined with any two of clarithromycin, a nitroimidazole and amoxycillin. These omeprazole triple therapy combinations provide eradication rates that are usually in the range of 85-95%, when assessed on a per protocol basis. Side-effects are minor and rarely interfere with compliance. Increasingly, these combinations are being given in a twice-daily dosage, making them more acceptable for the patient, and the dosage of antibiotics, in some cases, can be reduced. Furthermore, 1 week of treatment has been shown to be effective. In a few patients, however, even these highly effective eradication regimens fails, and anecdotal reports suggest that, once this has happened, other treatments are often similarly ineffective. Failure is not simply a matter of antibiotic resistance because patients with resistant organisms are often cured. In some patients, poor compliance, antibiotic resistance, coccoid bacterial forms, or the presence of sanctuary sites may be the cause of failure, in others, it has been suggested that pretreatment with an acid inhibitor may be the explanation. Research into these particular areas will be required, unless a new and universally effective drug combination can be identified. PMID- 8722390 TI - Cost-effectiveness of Helicobacter pylori eradication therapy in duodenal ulcer disease. AB - A model analysis applied to Helicobacter pylori eradication found that, following successful healing with omeprazole and H. pylori eradication, virtually all patients were cured and experienced no relapse during the next 5 years. In contrast, almost all of the patients receiving episodic therapy relapsed and, during maintenance therapy with H2-receptor antagonists, most experienced at least one relapse. Although H. pylori eradication initially resulted in higher costs than the alternative therapies, it reduced the risk of recurrence and, for most patients, no future costs were incurred. Even with a worst case scenario, such as an H. pylori eradication rate of only 50%, the H. pylori eradication therapy had a pay-off period of less than 1.3 years compared with maintenance treatment and 3 years compared with episodic treatment. A preliminary analysis also compared the cost-effectiveness of three different H. pylori eradication therapies: omeprazole plus one or two antibiotics, ranitidine plus two antibiotics, and ranitidine plus bismuth triple therapy. The highest eradication rates (in excess of 90%) were achieved using 1-week regimens including omeprazole in combination with either clarithromycin or amoxycillin and a nitroimidazole. These regimens were also shown to be the most cost-effective. As the difference in costs between the therapies is small compared with the savings that can be achieved by successful H. pylori eradication, it is logical that the eradication strategy with the highest eradication rate is the most cost-effective. The model analysis concludes that H. pylori eradication in patients with duodenal ulcer disease is cost-effective in comparison to episodic therapy with omeprazole or maintenance therapy with ranitidine. PMID- 8722391 TI - Helicobacter pylori reinfection after apparent eradication--the Ipswich experience. AB - The reported rate of Helicobacter pylori reinfection following eradication therapy is highly variable. In Ipswich, the 14C-urea breath test (UBT) has been used since 1986 as a tool to study H. pylori eradication and reinfection. Updated results from 1182 patients in whom the organism had apparently been successfully eradicated, following a number of different eradication regimens between October 1986 and 31 March 1995, are presented. During this period, 57 "reinfections' were observed, of which 45 had occurred within 6 months of treatment. After the first year, the 'reinfection' rate was less than 0.6% per year. The criterion for eradication of the infection was a UBT (2-hour area under curve) of less than 40 at least 1 month after treatment. The treatment regimens were arbitrarily divided into five groups with eradication rates of: less than 20%, 20-39%, 40-59%, 60-79% and over 80%. In these groups, the 6-month 'reinfection' rates were 28.0%, 15.8%, 16.4%, 4.6% and 1.7%, respectively (p < 0.001). These and other data presented in the paper strongly suggest that, in Westernized countries, most so-called reinfections in adults are in fact the late recrudescence of a suppressed infection rather than a true reinfection. Our data also suggest that the true reinfection rate is particularly low if the eradication therapy chosen has an efficacy of more than 85%. Several effective and well-tolerated 1-week triple H. pylori eradication regimens are now available, and we would advocate their use in preference to the less effective dual regimens where initial eradication rates are lower and there is consequently a higher risk of 'reinfection'. We would predict that even in developing countries with a high prevalence of metronidazole resistant H. pylori, the 'reinfection' rate would be low if a combination of omeprazole, amoxycillin and clarithromycin were to be used. PMID- 8722392 TI - Effects of permanent eradication or transient clearance of Helicobacter pylori on histology of gastric mucosa using omeprazole with or without antibiotics. AB - Changes in Helicobacter pylori-associated gastritis of the antrum and corpus were investigated in a large number of patients treated with omeprazole, with or without the addition of amoxycillin. To investigate the role of H. pylori associated gastritis in ulcerogenesis and its interplay with omeprazole, biopsies were taken and evaluated according to the Sydney system. Successful eradication of H. pylori (assessed histologically 4 weeks after the end of therapy) led to prompt and persistent suppression of gastritis activity, slow, partial regression of mononuclear inflammation and an ulcer recurrence rate of only 14% during the 6 months' follow-up. In patients treated with omeprazole and placebo, or where eradication treatment with omeprazole and amoxycillin had failed, transient clearance of H. pylori from the antral (but not oxyntic) mucosa was seen. In both of these groups of patients transient regression in the antrum, and worsening in the corpus, of gastritis activity and mononuclear inflammation were evident, coupled with ulcer recurrence rates of 72 and 46%, respectively. It was concluded that H. pylori colonization and gastritis activity play a crucial role in ulcerogenesis, that acid inhibition treatment improves antral H. pylori gastritis and worsens the oxyntic mucosal gastritis, and that this can be prevented by eradication of the H. pylori infection. PMID- 8722393 TI - Long-term consequences of Helicobacter pylori eradication: clinical aspects. AB - Helicobacter pylori is probably the most important factor in the pathogenesis of peptic ulcer disease in the absence of other precipitating factors, such as the intake of ulcerogenic drugs. Clinical studies have shown convincingly that eradication of H. pylori dramatically alters the long-term natural history of this chronic relapsing disorder, and that ulcer recurrences following eradication are rare. Available evidence also suggests that figures for H. pylori reinfection are low in adults in developed countries following eradication. In addition, H. pylori eradication may prevent peptic ulcer bleeding or rebleeding. Curing H. pylori infection significantly increases the quality of life of patients with duodenal ulcer disease and is, to date, the most cost-effective treatment in the long-term management of this disease. Ulcer patients presenting with recurrent dyspeptic symptoms after apparently successful eradication of H. pylori should be checked for ulcer and H. pylori recurrence, as well as for reflux oesophagitis, which has been shown to occur in about 9% of patients with duodenal ulcer disease and previously cured of H. pylori infection. We would also argue that H. pylori infection in young patients with dyspepsia should be treated, although this issue will remain controversial until well-designed, placebo-controlled studies establish the real benefits of eradication therapy in the long-term treatment of this disease. PMID- 8722394 TI - Managing patients in general practice. AB - General practitioners will, in the near future, have several choices of how to treat patients with peptic ulcer disease, as there will be many treatment guidelines that present different combinations of acid-inhibitory drugs, such as omeprazole, and antibiotics. However, they will still be left with the problem of who to treat, because patients do not present with a confirmed diagnosis; rather, they present with dyspeptic symptoms. PMID- 8722395 TI - Italian omeprazole triple therapy--a 1-week regimen. AB - Eradication of Helicobacter pylori infection is increasingly recognized as an appropriate therapeutic option, although it may prove difficult to achieve. Variable results, possibly due to side-effects, poor patient compliance and antimicrobial resistance, have been reported with traditional bismuth triple therapy. The combination of highly effective antisecretory drugs, such as omeprazole, and antimicrobial agents are providing a useful alternative to such traditional triple therapy. PMID- 8722396 TI - Helicobacter pylori--a revolution in understanding. Proceedings of 2 symposia. Osaka and Tokyo, 27 and 28 November 1994. PMID- 8722397 TI - History of Helicobacter infection. AB - Helicobacter pylori infection is now recognized as the cause of most cases of chronic gastritis, and a major causal factor in peptic ulcer disease. However, though the presence of intragastric bacteria has been recognized for many years, the connection between H. pylori and peptic ulcers has only been established during the last decade. Pathological studies have suggested that intragastric bacteria damage the gastric mucosa, reducing its resistance to acid, while epidemiological studies have suggested that a non-physiological factor, such as bacterial infection, is involved in the development of peptic ulcer. Subsequently, H. pylori, associated with chronic gastritis, was identified in gastric biopsies from patients with peptic ulcer. Eradication of H. pylori has been shown to reduce the recurrence of peptic ulcer after ulcer healing, and such treatment is now widely accepted. PMID- 8722398 TI - The nature of Helicobacter pylori. AB - Helicobacter pylori is an important pathogen in humans, causing chronic gastritis and playing a major role in the development of peptic ulcers and gastric cancer. The organism is highly adapted to the human stomach, largely due to its motility and ability to produce large amounts of urease. It binds specifically to the gastric mucosa via adhesion pedestals; colonization of the duodenum only occurs in the presence of gastric metaplasia. Infection with H. pylori leads to gastritis, but the majority of infected patients are asymptomatic, and it is thought that the ability of H. pylori to cause more severe disease may be related to the presence of the cagA gene. With improvements in public health and living conditions, the prevalence of H. pylori infection in developed countries is decreasing, and this is associated with a decline in the incidence of peptic ulcer and gastric cancer. PMID- 8722399 TI - Helicobacter pylori and the evolution of gastritis. AB - Following the isolation of Helicobacter pylori from the stomachs of patients with peptic ulcer in 1982, intensive research has examined the relationship between this organism and gastritis. H. pylori affects the grade and activity of gastritis, the quality of surface epithelial cells, and the nature of mucus production. Gastritis results from a direct action, involving bacterial cytotoxins and enzymes, and also in an indirect way through the effect of monocyte and granulocyte infiltration. Eradication of H. pylori leads to the long term healing of gastritis. H. pylori is also associated with intestinal metaplasia and the development of mucosa-associated lymphoid tissue (MALT). Regression of MALT lymphomas has been reported after H. pylori eradication treatment. PMID- 8722400 TI - Gastritis in the Japanese stomach. AB - Endoscopic studies of Japanese patients with gastritis have revealed the existence of an endoscopic atrophic border that marks the transition between non atrophic gastritis and atrophic gastritis, and between fundic glands and pyloric glands. Marked changes in cell type, gastritis activity, atrophy and density of Helicobacter pylori infection occur across this border. Gastritis appears to extend from the antrum to the corpus more quickly in Japanese patients than in patients from other populations. Infection with H. pylori may be an important factor influencing the rate of progression of gastritis in Japanese patients. PMID- 8722401 TI - Helicobacter pylori and peptic ulcer disease. AB - Infection with Helicobacter pylori is a causal factor in the development of peptic ulcers. There is a strong association between gastritis induced by H. pylori and ulcer development, and peptic ulcer disease seldom develops in the absence of infection. Eradication of H. pylori leads to long-term remission and cure of peptic ulcer disease. The ability of H. pylori to cause ulcers appears to be linked to strain-specific characteristics, such as the presence of the cagA gene. Studies of ulcer patients with healed H. pylori infection suggest that the pathogenesis of peptic ulcers in Japan is comparable to that elsewhere in the world. Smoking, a recognized risk factor for peptic ulcer disease, does not cause ulcers in the absence of infection with H. pylori; by contrast, the presence of H. pylori is not necessary for the development of ulcers associated with non steroidal anti-inflammatory drugs (NSAIDs). Reinfection after eradication of H. pylori occurs at a rate similar to the infection rate; both are infrequent events in adults living in industrialized countries. Effective and well-tolerated eradication therapies are now available, and represent the most cost-effective treatment for peptic ulcer disease. PMID- 8722402 TI - Helicobacter pylori and dyspepsia. AB - Non-ulcer dyspepsia (NUD) is a common condition that often goes unrecognized. Although approximately 50% of patients with NUD are infected with Helicobacter pylori, attempts to establish a link between this organism and the pathogenesis of NUD have yielded conflicting results. Recent studies have shown that eradication of H. pylori leads to improvements in symptoms in patients with NUD, and have highlighted the importance of long-term follow-up in this condition. It is possible that patients with NUD and H. pylori infection have a discrete condition that forms part of a spectrum of peptic ulcer disease. Hence, it may soon be possible to recommend H. pylori eradication treatment for patients with NUD. PMID- 8722403 TI - Helicobacter pylori and gastric cancer. AB - Epidemiological studies have consistently shown an association between infection with Helicobacter pylori and gastric cancer. The strongest evidence comes from three prospective studies that have shown the risk of gastric cancer to be increased by about fourfold in infected individuals. This raises the possibility of preventing a substantial number of gastric cancer cases by controlling H. pylori. Screening asymptomatic populations and eradicating the organism in those infected is the only treatment strategy currently available, but several questions remain to be answered before this can be recommended as a routine public health measure. Research into H. pylori has transformed understanding of the aetiology of gastric cancer, focusing attention on factors such as chronic inflammation, impaired host defence mechanisms and oxidative cell damage. PMID- 8722404 TI - The pathological sequence in the development of gastric cancer: I. AB - Although Helicobacter pylori has been designated a Group 1 carcinogen in humans, a causal role for this organism in the pathogenesis of gastric cancer has yet to be established. Histogenetically, gastric cancers have been classified into two main histological types, diffuse and intestinal, which have differing characteristics and pathogenesis. In diffuse-type cancer, minute signet ring tumours develop de novo in the neck region of glands. Gastritis caused by H. pylori infection does not contribute to the development of gastric cancer in this situation. In intestinal-type gastric cancer, the primary event is not always intestinal metaplasia, and the significance of H. pylori in this condition is not clear. There is evidence that a proportion of adenocarcinoma initially have a gastric-type phenotype, but undergo transformation to the intestinal type as the condition progresses. PMID- 8722405 TI - The pathological sequence in the development of gastric cancer: II. AB - Studies of gastric microcarcinomas (size: 5 mm or less in diameter) have highlighted the importance of intestinal metaplasia in the development of gastric cancer. In this study, a consecutive selection of 57 microcarcinomas were examined and classified: 16 were gastric-phenotype tumours and 41 were predominantly intestinal-phenotype tumours. Intestinal metaplasia was severe within the surrounding mucosa of 31 of these intestinal-type carcinomas, moderate in 7 and mild in 3. Residual (pseudo) pyloric glands were found below the centre of 22 of the 41 intestinal-type tumours, but the centre of the remaining 19 tumours occupied the full thickness of the mucosa. It is suggested that intestinal-type microcarcinomas develop in the intestinalized mucosa or in the double-phenotypic mucosa (i.e. with intestinal type in the upper part and gastric type in the lower part). Understanding the relationship between intestinal metaplasia and gastric cancer may help to clarify the involvement of Helicobacter pylori in gastric cancer. PMID- 8722406 TI - Diagnosis of Helicobacter pylori infection. AB - A number of invasive and non-invasive techniques are available for the diagnosis of infection with Helicobacter pylori. These can be used to establish whether infection is present and to confirm that eradication has been achieved after treatment. Invasive techniques require biopsy of the gastric mucosa; such techniques include histology, bacterial culture, the rapid urease test and the polymerase chain reaction (PCR) technique. The principal non-invasive techniques are the 13C-urea breath test and serological assays in serum or saliva samples. Histological examination of gastric biopsy samples should be mandatory at the initial presentation of the patient because it also gives insight on the status of the gastric mucosa. The rapid urease test can be used to give a rapid diagnosis of infection. Eradication should also be confirmed by histological examination, though other sensitive tests such as bacterial culture, PCR or the 13C-urea breath test may also be used. PMID- 8722407 TI - Eradication of Helicobacter pylori. AB - It has become clear over the past 10 years that Helicobacter pylori is responsible for two of the most common gastrointestinal diseases worldwide, gastric and duodenal ulcer, and that it plays an important role in the development of the second most common lethal malignancy affecting humankind, gastric cancer. At present eradication therapy is considered to be the most appropriate way of treating gastric and duodenal ulcer associated with infection. Treatment, however, remains a difficult area. Although the organism is sensitive to a wide variety of antibiotics in vitro, monotherapy in vivo is rarely successful. The first treatments used bismuth compounds, but results were variable and side-effects significant. When a single antibiotic, such as amoxycillin or clarithromycin, is combined with the proton pump inhibitor omeprazole, results are good but still with varied success and in the case of clarithromycin, the organism can develop resistance to macrolides. Triple therapy using omeprazole, 20 mg daily, in conjunction with clarithromycin, 250 mg twice daily, and tinidazole, 500 mg twice daily, and given for only one week, has had a 95% success rate. This regimen appears to be the most effective and acceptable combination described so far. However, the major difficulty facing most clinicians today is that even with the most effective therapies, infection in some patients would appear to be ineradicable, and this is an area in which further research is urgently required. PMID- 8722408 TI - Suppression of Helicobacter pylori colonization with omeprazole. AB - The efficacy of omeprazole, 20 mg once daily, in suppressing Helicobacter pylori colonization was studied in 33 patients with peptic ulcers. Omeprazole treatment produced a significant fall in the number of H. pylori colony-forming units in the antrum. This finding, together with the results of a meta-analysis of clinical trials of H. pylori eradication therapy, suggests that combinations of omeprazole and antibiotics may be useful as eradication therapy in patients with peptic ulcer disease and H. pylori infection. PMID- 8722409 TI - Treatment of Helicobacter pylori infection by topical administration of antimicrobial agents. AB - A technique has been developed in which a combination of bismuth subnitrate, amoxycillin and metronidazole is instilled into the stomach via a nasogastric tube and removed after 1 hour, thereby avoiding the side-effects often seen with triple therapy. This technique has achieved eradication of H. pylori in 24 of 25 patients with non-ulcer dyspepsia. A modified version of the technique was used in six patients, and eradication achieved in each case. PMID- 8722410 TI - Helicobacter pylori in Japan. AB - Recent international consensus statements have concluded that Helicobacter pylori is a causal factor in peptic ulcer disease and a Group 1 carcinogen in humans, and that all patients with peptic ulcer associated with H. pylori infection should receive eradication therapy. There are marked differences in the presentation of peptic ulcer disease between Japanese patients and those from other countries, however, and thus a committee of the Japanese Society of Gastroenterology has been set up to develop guidelines for clinical trials in patients with gastric or duodenal ulcers associated with H. pylori. According to these guidelines, eradication therapy should normally consist of dual therapy with a proton pump inhibitor, such as omeprazole, and an antibiotic; a nitroimidazole can be added, however, if necessary. The diagnosis of H. pylori should be made on the basis of a positive culture of the organism or histological examination of gastric biopsies, together with a positive urease test or 13C-urea breath test. Eradication should be confirmed by the same means 4-6 weeks after treatment. Other research is investigating the relationship between H. pylori and gastric cancer, an important issue in view of the high incidence of this condition in Japan. There is evidence that the Mongolian gerbil may be a useful animal model to investigate this question. PMID- 8722411 TI - The coronary artery disease paradox. AB - After effective antihypertensive therapy was introduced in the 1950s, we rapidly showed that lowering blood pressure in patients with hypertensive crises saved lives. In the 1960s, the benefit for those with asymptomatic elevations of diastolic blood pressure levels of > or = 115 mm Hg was proven and, in the early 1970s, the value for those with diastolic levels ranging from 105 to 114 mm Hg was demonstrated. In the early 1990s, the Systolic Hypertension in the Elderly Program (SHEP) showed benefits for elderly hypertensive patients with isolated systolic hypertension. Because it has been difficult to show that treating hypertensive patients with diastolic elevations from 90 to 99 mm Hg reduces clinical events, metaanalyses were used to pool available clinical trial data to evaluate antihypertensive therapy for such patients. Using this technique, it was clear that active therapy reduced strokes as expected but fell short for coronary artery disease. There are many possible explanations for this "coronary artery disease paradox," including the misclassification and stratification of hypertensive patients, inadequacy of available trial data, unidentified and identified competing cardiovascular risk factors, and limited selection of the antihypertensive agents used in the trials analyzed, which were exclusively diuretics and beta-blockers. This paper will review these possible reasons for the coronary artery disease paradox, concentrating on classification and problems with drug therapy. PMID- 8722412 TI - Chronobiology and chronotherapeutics. Applications to cardiovascular medicine. AB - The concept of homeostasis, ie, constancy of the milieu interne, has dominated the teaching, research, and practice of medicine during the 20th century. According to this theory, biologic functions and processes are maintained in relative constancy over time. The emerging concepts of chronobiology, the scientific discipline of biologic rhythm study, and the findings from research in this field challenge the construct of homeostasis. Epidemiology studies document that the occurrence and exacerbation of many human diseases vary predictably in time over 24 h, the menstrual cycle, and year due to underlying rhythms of corresponding period. Advances in the chronobiology of cardiovascular disease have proceeded rapidly during the past decade and have influenced the manner in which diagnostic procedures are conducted and interpreted. Twenty-four-hour ambulatory blood pressure (BP) monitoring and Holter monitoring reveal the marked circadian (24-h) rhythms in BP in hypertensive patients and electrocardiographic events in patients with ischemic heart disease. Homeostatically devised pharmacotherapies, ie, medications designed to ensure constant drug levels over time, may be inadequate to optimally control diseases whose courses vary in risk or severity during the 24-h period. Chronotherapies--medications formulated to deliver varying amounts of drug at different times during the 24 h period to correlate with biologic need--theoretically could offer improved efficacy. A chronotherapy for cardiovascular disease already exists in the form of the evening administration for lipid-lowering medications. The chronotherapy for hypertension and of ischemic heart disease is forthcoming. PMID- 8722413 TI - The pathophysiology of the onset of morning cardiovascular events. AB - Evidence obtained over the past decade indicates that myocardial infarction (MI) and sudden death are not random events but rather, in many cases, may be triggered by the daily activities of the subject. The importance of physical or mental stresses as triggers is suggested by the parallel morning increased onsets of MI, sudden cardiac death, and stroke. Unstable angina and MI are usually precipitated by thrombus formation over a disrupted plaque that causes partial or complete obstruction of coronary artery blood flow. This process may be caused by physiologic factors that lead to rupture of a vulnerable plaque and subsequent thrombosis. Beta-blockers and aspirin, which can diminish these physiologic processes, have been shown to blunt or abolish the morning peak of onset of acute MI. It is hypothesized that occlusive coronary thrombosis occurs when an atherosclerotic plaque becomes vulnerable to rupture, and mental or physical stress causes the plaque to rupture. Increases in coagulability or vasoconstriction triggered by daily activities may also contribute to complete occlusion of the coronary artery lumen. Recognition of the circadian variation of the onset of acute cardiovascular disease suggests the need for pharmacologic protection of patients during the vulnerable periods and provides clues to the mechanism of disease onset, the investigation of which may lead to improved methods of prevention. PMID- 8722414 TI - A chronotherapeutic approach to the management of hypertension. AB - Patients with hypertension have a characteristic circadian pattern of blood pressure (BP) variability in a 24-h period, characterized by a wide range while awake and active, and a narrower range during sleep and inactivity. Blood pressure in the untreated hypertensive individual declines by approximately 10% to 20% during sleep compared with the mean awake BP values. Early in the morning (assuming a typical day-night activity cycle), the BP rises sharply on awakening, when physical and mental activities increase. Antihypertensive therapy has been traditionally dosed in the morning after awakening, and in recent years most of the newly developed antihypertensive agents have been once-daily, long-acting preparations. However, theoretically, this pattern of drug dosing may be suboptimal since pharmacodynamic activity is attenuated at the end of the dosing period, when a large rise in BP may occur. Presently, only a few studies have been performed to evaluate the effects of nocturnal dosing of standard long acting drugs. Consequently, we undertook an assessment of the effects of a new formulation of verapamil--controlled onset extended release verapamil HCl (COER 24)--that is dosed nocturnally and has a controlled onset of delivery (4 to 5 h postdose) and an extended release for the remainder of the dosing period. Through ambulatory BP monitoring, evaluation revealed that this verapamil formulation produced changes in BP that followed the circadian rhythm of BP: lower reductions during sleep, when the BP is intrinsically lowest, and appropriately larger reductions during the early morning and daytime hours, when ambulatory BP values accelerate and plateau to their highest levels. These data demonstrate that it is possible to design antihypertensive therapy for once-nightly dosing, thereby providing a chronotherapeutic regimen for patients with hypertension. PMID- 8722415 TI - The emerging role of chronotherapeutics in managing hypertension. Panel discussion. PMID- 8722416 TI - Monoclonal antibodies in tumor therapy. PMID- 8722417 TI - Experimental radioimmunotherapy: biological effectiveness and comparison with external beam radiation. PMID- 8722418 TI - Higher efficiency of 131I-labeled anti-carcinoembryonic antigen-monoclonal antibody F(ab')2 as compared to intact antibodies in radioimmunotherapy of established human colon carcinoma grafted in nude mice. PMID- 8722419 TI - Problems and advances in the dosimetry of radionuclide targeted therapy. PMID- 8722420 TI - Minimal residual disease: a target for radioimmunotherapy with 131I-labeled monoclonal antibodies? Some dosimetric considerations. PMID- 8722421 TI - Optimal therapeutic strategies for radioimmunotherapy. PMID- 8722423 TI - Can preirradiation enhance tumor uptake of radiolabeled pharmaceuticals? Experimental data in a mouse neuroblastoma xenograft system. PMID- 8722422 TI - Two approaches for enhancing radioimmunotherapy: alpha emitters and hyperthermia. PMID- 8722424 TI - The use of radioimmunotherapy in combination with bioreductive agents. PMID- 8722426 TI - The treatment of patients with recurrent malignant gliomas with intratumoral radioimmunoconjugates. PMID- 8722425 TI - Treatment of diffuse leptomeningeal malignancy by intrathecal injection of 131I radioimmunoconjugates. PMID- 8722427 TI - High dose radioimmunotherapy in malignant lymphoma. PMID- 8722428 TI - The use of epidermal growth factor receptor-425 monoclonal antibodies radiolabeled with iodine-125 in the adjuvant treatment of patients with high grade gliomas of the brain. PMID- 8722429 TI - A telecommunications system for monitoring and counseling patients with hypertension. Impact on medication adherence and blood pressure control. AB - This study was conducted to evaluate the effect of automated telephone patient monitoring and counseling on patient adherence to antihypertensive medications and on blood pressure control. A randomized controlled trial was conducted in 29 greater Boston communities. The study subjects were 267 patients recruited from community sites who were >or= 60 years of age, on antihypertensive medication, with a systolic blood pressure (SBP) of >or= 160 mm Hg and/or a diastolic blood pressure (DBP) of >or= 90 mm Hg. The study compared subjects who received usual medical care with those who used a computer-controlled telephone system in addition to their usual medical care during a period of 6 months. Weekly, subjects in the telephone group reported self-measured blood pressures, knowledge and adherence to antihypertensive medication regimens, and medication side effects. This information was sent to their physicians regularly. The main study outcome measures were change in antihypertensive medication adherence, SBP and DBP during 6 months, satisfaction of patient users, perceived utility for physicians, and cost-effectiveness. The mean age of the study population was 76.0 years; 77% were women; 11% were black. Mean antihypertensive medication adherence improved 17.7% for telephone system users and 11.7% for controls (P = .03). Mean DBP decreased 5.2 mm Hg in users compared to 0.8 mm Hg in controls (P = .02). Among nonadherent subjects, mean DBP decreased 6.0 mm Hg for telephone users, but increased 2.8 mm Hg for controls (P = .01). For telephone system users, mean DBP decreased more if their medication adherence improved (P = .03). The majority of telephone system users were satisfied with the system. Most physicians integrated it into their practices. The system was cost-effective, especially for nonadherent patient users. Therefore, weekly use of an automated telephone system improved medication adherence and blood pressure control in hypertension patients. This system can be used to monitor patients with hypertension or with other chronic diseases, and is likely to improve health outcomes and reduce health services utilization and costs. PMID- 8722430 TI - Blood pressure evaluation by noninvasive and traditional methods. Consistencies and discrepancies among photoplethysmomanometry, office sphygmomanometry, and ambulatory monitoring. Effects of blood pressure measurement. AB - Three hundred eight outpatients referred for hypertension were studied. A continuous beat-to-beat noninvasive recording (Finapres) of blood pressure evaluated the blood pressure increase (9 mm Hg systolic and 4 mm Hg diastolic) induced by office sphygmomanometry. Thereafter, patients underwent a 24-h ambulatory blood pressure monitoring. The evaluation against Finapres showed that office sphygmomanometry overestimates the systolic blood pressure by 3 +/- 36 mm Hg (mean +/- 2 SD) and the diastolic blood pressure by 15 +/- 25 mm Hg (mean +/- 2 SD). Blood pressure monitoring showed similar discrepancies. On the basis of both monitoring (normalcy defined from a population of 550 normotensive subjects) and office sphygmomanometry, patients were considered normotensive, hypertensive (either untreated or under active drug treatment), white coat hypertensive (monitoring below the 95th percentile and sphygmomanometry more than 140/90 mm Hg, either untreated or under active drug treatment), and reverse white coat patients (monitoring over the 95th percentile and sphygmomanometry less than 140/90 mm Hg). Patients showed different levels of alert reaction (the highest in white coat hypertensive and the lowest in reverse white coat hypertensive patients), and a similar increase in blood pressure induced by conventional sphygmomanometry. During initial readings of ambulatory monitoring, blood pressure decreased from the first reading to the third reading. This decrease is related to the increase of blood pressure under sphygmomanometry. Caution should be paid in interpreting results of sphygmomanometry (error level in the single patient as high as +/- 40 mm Hg), and interpreting and averaging results of the first hour of blood pressure monitoring (variably affected by the alert reaction to the clinical environment). PMID- 8722431 TI - Circadian variations of blood pressure in patients with sequelae of carbon monoxide poisoning. AB - It has been shown that carbon monoxide poisoning causes necrosis of the globus pallidum and the cerebral cortex, and a diffuse demyelination of the cerebral subcortical white matter, resulting in the impairment of the higher brain functions manifested as memory disturbances, apraxia, and agnosia. The purpose of the present study was to determine the effects of the lesions in the brain caused by carbon monoxide poisoning on the circadian changes in blood pressure and pulse rate. We measured the ambulatory blood pressure in 15 male patients with the sequelae of acute carbon monoxide poisoning and 16 age- and sex-matched controls. Using either brain computed tomography or brain magnetic resonance imaging, we determined that seven patients had lesions in the globus pallidum bilaterally, four had lesions in the parietotemporooccipital lobe bilaterally, and five had multiple deep white matter lesions. Circadian variations of blood pressure and pulse rate did not show any significant differences in either group. The average 24-h blood pressures were 120.8 +/- 2.2 (systolic)/74.1 +/- 1.5 mm Hg (diastolic) in the patients and 117.4 +/- 2.7/74.5 +/- 2.1 mm Hg in the controls. The daytime (6:00 to 21:00) and nighttime (21:00 to 6:00) blood pressures were 127.3 +/- 2.3/78.0 +/- 1.5 mm Hg and 109.7 +/- 2.4/67.4 +/- 1.7 mm Hg in the patients, and 121.6 +/- 2.9/77.5 +/- 2.2 mm Hg and 110.0 +/- 2.7/69.3 +/- 1.8 mm Hg in the controls, respectively. Furthermore, there were no differences in cardiovascular and plasma catecholamine responses induced by either a head-up tilt or a cold pressor test between the two groups. It is concluded that diffuse or multiple lesions in bilateral cerebral hemispheres caused by carbon monoxide poisoning per se do not affect the circadian changes in blood pressure and pulse rate observed in normotensive subjects. PMID- 8722432 TI - Efficacy and safety of fosinopril/hydrochlorothiazide combinations on ambulatory blood pressure profiles in hypertension. Fosinopril/Hydrochlorothiazide Investigators. AB - Twenty-four-hour ambulatory blood pressure monitoring (ABPM) was used to assess the antihypertensive efficacy and safety of the angiotensin converting enzyme (ACE) inhibitor fosinopril (Fos) in combination with hydrochlorothiazide (HCTZ) in doses of 10/12.5 mg and 20/12.5 mg taken once daily versus placebo in patients with mild-to-moderate hypertension. In two methodologically identical studies, the antihypertensive effects were evaluated by 24-h ABPM and by seated office diastolic (DBP) and systolic (SBP) blood pressures. After a 4- or 5-week placebo washout, 79 patients received randomized, double-blind treatment for 8 weeks with either the Fos/HCTZ 10/12.5-mg dose combination (n = 41) or placebo (n = 38), and in the second study, 62 patients were treated with either the Fos/HCTZ 20/12.5-mg dose combination (n = 30) or placebo (n = 32). Changes from baseline in mean 24-h systolic and diastolic ABPM were significantly different from placebo for both doses (SBP/DBP with 10/12.5 mg, -18.2/ -10.1 mm Hg, P or= 30 mg/24 h (overt microalbuminuria). Office systolic BP was not different in the three groups, whereas 24-h systolic BP was higher in the subjects with microalbuminuria than in those with normal AER (P < .0001) and was similar in the two microalbuminuric groups. Office and 24-h diastolic BPs were higher in the subjects with overt microalbuminuria than in those with normal AER. Left ventricular mass was correlated to systolic (P < .0001) and diastolic (P = .01) 24-h BP, but was unrelated to AER. Family history for hypertension, smoking, coffee and alcohol intake, and physical activity habits did not influence AER. In a logistic regression analysis, 24-h systolic BP emerged as the only determinant of microalbuminuria (P < .0001). In conclusion, these results indicate that borderline levels of microalbuminuria may also be clinically relevant in stage I hypertension. Overweight and lifestyle factors do not appear to influence AER in these patients. Finally, the lack of correlation between AER and left ventricular mass suggests that renal and cardiac involvement do not occur in a parallel fashion in the initial phase of hypertension. PMID- 8722437 TI - Rationale and design for the Antihypertensive and Lipid Lowering Treatment to Prevent Heart Attack Trial (ALLHAT). ALLHAT Research Group. AB - Are newer types of antihypertensive agents, which are currently more costly to purchase on average, as good or better than diuretics in reducing coronary heart disease incidence and progression? Will lowering LDL cholesterol in moderately hypercholesterolemic older individuals reduce the incidence of cardiovascular disease and total mortality? These important medical practice and public health questions are to be addressed by the Antihypertensive and Lipid Lowering Treatment to Prevent Heart Attack Trial (ALLHAT), a randomized, double-blind trial in 40,000 high-risk hypertensive patients. ALLHAT is designed to determine whether the combined incidence of fatal coronary heart disease (CHD) and nonfatal myocardial infarction differs between persons randomized to diuretic (chlorthalidone) treatment and each of three alternative treatments--a calcium antagonist (amlodipine), an angiotensin converting enzyme inhibitor (lisinopril), and an alpha-adrenergic blocker (doxazosin). ALLHAT also contains a randomized, open-label, lipid-lowering trial designed to determine whether lowering LDL cholesterol in 20,000 moderately hypercholesterolemic patients (a subset of the 40,000) with a 3-hydroxymethylglutaryl coenzyme A (HMG CoA) reductase inhibitor, pravastatin, will reduce all-cause mortality compared to a control group receiving "usual care." ALLHAT's main eligibility criteria are: 1) age 55 or older; 2) systolic or diastolic hypertension; and 3) one or more additional risk factors for heart attack (eg, evidence of atherosclerotic disease or type II diabetes). For the lipid-lowering trial, participants must have an LDL cholesterol of 120 to 189 mg/dL (100 to 129 mg/dL for those with known CHD) and a triglyceride level below 350 mg/dL. The mean duration of treatment and follow-up is planned to be 6 years. Further features of the rationale, design, objectives, treatment program, and study organization of ALLHAT are described in this article. PMID- 8722438 TI - Presence of cardiovascular structural changes in essential hypertensive patients with coronary microvascular disease and effects of long-term treatment. AB - In asymptomatic essential hypertensive patients with angiographically normal coronary arteries and without left ventricular hypertrophy, dipyridamole-induced ischemic-like ST segment depression may be a marker of coronary microvascular disease. In this study we evaluated, first, whether this cardiac abnormality is linked to structural or functional vascular abnormalities, and second, the effect of antihypertensive treatment by 12-month administration of the angiotensin converting enzyme (ACE) inhibitor captopril (50 mg twice a day orally). In essential hypertensives with dipypridamole echocardiography stress test (DET) (DET+, n = 8) and without (DET-, n = 8) ST segment depression greater than 0.1 mV during intravenous dipyridamole infusion (0.84 mg/kg over 10 min), we studied the forearm blood flow (FBF, venous plethysmography, mL/100) modifications induced by intrabrachial acetylcholine (Ach) (0.15, 0.45, 1.5, 4.5, 15 micrograms/100 mL/min x 5 min each), an endothelium-dependent vasodilator, and by sodium nitroprusside (SNP) (1, 2, 4 micrograms/100 mL/min x 5 min each), a smooth muscle cell relaxant compound. Minimal forearm vascular resistances (MFVR), an index of arteriolar structural changes, were also calculated. Both Ach and SNP caused greater vasodilation in DET- as compared to DET+ while MFVRs were lower in DET- compared to DET+. After treatment, both DET+ and DET- patients showed a significant and similar reduction in blood pressure and left ventricular mass index, while vasodilation to acetylcholine and sodium nitroprusside was increased only in the DET+ group. In addition, forearm minimal vascular resistances were significantly reduced only in DET+ patients, who showed disappearance of dipyridamole-induced ischemic-like ST segment depression. In conclusion, these data confirm that essential hypertensive patients with microvascular coronary disease are characterized by the presence of structural changes in the forearm vascular bed. Our results also indicate that both cardiac and forearm vascular abnormalities can be reversed by antihypertensive treatment with an ACE inhibitor. PMID- 8722439 TI - Defective endothelium-dependent relaxation in fructose-hypertensive rats. AB - The present study examined the endothelium-dependent and -independent responses of isolated mesenteric arteries to acetylcholine and the endothelium-independent vasodilator sodium nitroprusside in mesenteric arteries from fructose-induced hypertensive rats. Fructose feeding resulted in hyperinsulinemia and elevated blood pressure when compared to controls (plasma insulin, 5.9 +/- 0.4 v control 3.6 +/- 0.4 ng/mL, P < .05; systolic blood pressure, 154 +/- 5 v control 127 +/- 7 mm Hg, P < .05). The maximum contractile response of mesenteric arteries to norepinephrine did not differ between the control and fructose groups, either with or without the endothelium. In arteries with intact endothelia, precontracted with the approximate ED50 of norepinephrine, the percent maximum relaxation produced by acetylcholine in hypertensive rats was lower than the control arteries (62 +/- 7 v control 95 +/- 5, P < .05) without any change in sensitivity. In arteries precontracted with norepinephrine, the endothelium independent vasodilator sodium nitroprusside produced a dose-dependent relaxation in arteries obtained from control and fructose groups, both with and without the endothelium. The maximum relaxation produced by sodium nitroprusside did not differ between control and fructose arteries, either with or without the endothelium; however, removal of the endothelium caused an increase in sensitivity of this agonist. These data suggest that in the insulin resistant and hyperinsulinemic fructose-hypertensive rats, there is a defective endothelium dependent yet preserved endothelium-independent relaxation. PMID- 8722441 TI - Different vascular responses to subpressor angiotensin II administration in the mesenteric and renal circulation of rats. AB - Administered in small doses, angiotensin II (ANG II) potentiates its own pressor responses (autopotentiation). In the present study, we investigated whether potentiation of vasoconstrictor responses in the mesenteric and renal circulation accounted for the autopotentiation of pressor responses. Male Sprague-Dawley rats, 350 to 400 g, were infused with 200 ng/kg/min ANG II intraperitoneally for 7 to 10 days. Control rats were fitted with an osmotic minipump containing vehicle only. Vascular responses to ANG II, norepinephrine (NE), arginine vasopressin (AVP) and nerve stimulation were measured in the mesenteric and renal circulation of rats. In ANG II-treated rats: 1) tail systolic BP was 4 mm Hg higher than in controls (P = NS); 2) mesenteric vascular responses to ANG II (P < .002) and nerve stimulation (P = .03) were increased, and those to NE and AVP were unchanged; and 3) renal vascular responses to nerve stimulation were reduced (P < .02), and those to ANG II, NE, and AVP were unchanged. Thus, we demonstrated autopotentiation of vascular responses to ANG II in the mesenteric, but not the renal circulation. An interaction between ANG II and sympathetic activity may explain these differences in regional vascular responses. Regional hemodynamic measurements may provide important clues to pathogenetic mechanisms in hypertension. PMID- 8722440 TI - Activation of nitric oxide synthesis in vascular smooth muscle cells and macrophages during development in spontaneously hypertensive rats. AB - We previously showed that the lymphocyte proliferation response was significantly suppressed in spontaneously hypertensive rats (SHR) and that this depressed response was due to excessive production of nitric oxide (NO) in macrophages and vascular smooth muscle cells (VSMC). Whether lymphocyte depression and activation of NO synthesis are related to age and development of hypertension remains unclear. The present study addresses such a correlation by examining the time course of development of hypertension, NO synthesis alteration and lymphocyte depression in SHR. Our results show that 1) SHR spleen cell proliferation responses are depressed at 4, 8, and 12 weeks and 1 year of age, with the lowest response occurring at 4 weeks of age; 2) this depressed response is corrected by either NO synthase inhibitor or removal of macrophages from spleen cells; 3) NO production by SHR spleen macrophages is significantly higher in all age groups; 4) upon stimulation with lipopolysaccharide or cytokines, SHR VSMC produce a significantly greater amount of NO in all age groups; 5) the increase in NO synthesis in VSMC correlates significantly with the rise in blood pressure in SHR. However, statistical correlation analysis suggests that lymphocyte depression and the alteration of NO synthesis in macrophages were not associated with either age or increased blood pressure in SHR. On the contrary, the activation of NO synthesis in VSMC can be statistically correlated with elevated blood pressure throughout the development of hypertension in SHR. Nevertheless, the results also suggest that a general alteration in the NO synthesis system may exist in SHR. PMID- 8722442 TI - Influence of pressure overload and ACE inhibitor therapy on constitutive protein mRNA expression in the spontaneously hypertensive rat. AB - Despite use as constitutive protein standards to quantify mRNA, data are limited regarding alteration of cyclophilin or glyceraldehyde-3-phosphate dehydrogenase (G3PDH) in hypertension or angiotensin converting enzyme (ACE) inhibitor treatment. We assessed these standards in 6 month old Wistar-Kyoto rats (WKY, n = 16), compared to age-matched spontaneously hypertensive rats (SHR, n = 14). Additional SHR (n = 8) had received enalapril for 3 to 4 months at evaluation. Left ventricular (LV) and kidney RNA was extracted for dot blot cyclophilin and G3PDH cDNA hybridization. Cyclophilin and G3PDH mRNA densitometries were expressed as a ratio. Cyclophilin/G3PDH for the WKY, untreated SHR, and enalapril SHR were 1.56 +/- 0.33, 1.45 +/- 0.42, and 1.49 +/- 0.51, respectively, for the LV, and 1.52 +/- 0.09, 1.43 +/- 0.22, and 1.38 +/- 0.22, respectively, for the kidney. Differences were not significant. Relative expression of cyclophilin/G3PDH was unaffected by genetic SHR hypertension, or long term enalapril. Thus, either constitutive mRNA may be confidently used to index structural or functional protein responses, at the transcriptional level, in the SHR. PMID- 8722443 TI - American Society of Hypertension Workshop on Absolute Risk. PMID- 8722444 TI - Dopamine receptor signaling defects in spontaneous hypertension. AB - Dopamine produced by renal proximal tubules acts as an intrarenal natriuretic factor by direct tubular action; this paracrine effect is influenced by the state of sodium balance. Up to 60% of sodium excretion with volume (2%-10%) expansion may be mediated by D1-like receptors. The renal paracrine effect of dopamine is impaired in genetic hypertension; this is due to defects in renal dopamine production or transduction of the dopamine signal. The Dahl salt sensitive rat and the spontaneously hypertensive rat (SHR), which have normal renal dopamine production and expression of dopamine receptors, have a defect in the coupling of a D1-like receptor to G-protein/effector enzyme complex. A consequence of the defective D1-like receptor/effector enzyme coupling in SHR is a decreased ability of D1 agonists to inhibit Na+/H+ exchange and Na+/K+-ATPase activity. The defect is 1) genetic, since it precedes the onset of and cosegregates with the hypertension; 2) receptor specific, since it is not shared by other humoral agents; and 3) confined to the renal proximal tubule. Two of the cloned dopamine receptors in mammals are D1-like (D1A and D1B). The D1A receptor gene is expressed to a greater extent in renal proximal tubules than the D1B receptor gene. The D1-like receptor is important in the pathogenesis of hypertension. Chronic blockade of dopamine receptors accelerates the development of hypertension in normotensive and hypertensive rats. Moreover, disruption of the D1A receptor gene in mice increases systolic blood pressure and results in diastolic hypertension. The abnormal D1-like receptor in SHR may be the D1A receptor; its uncoupling from the G-protein/effector enzyme complex in renal proximal tubules of SHR may be due to mistargeting. The mechanism for this "mistargeting" of the D1A receptor is not due to a mutation in the primary sequence and remains to be determined. PMID- 8722445 TI - The heart in hypertension. Thinking small. PMID- 8722446 TI - ALLHAT: the largest and most important clinical trial in hypertension ever done in the USA. Antihypertensive and Lipid Lowering Treatment to Prevent Heart Attack Trial. PMID- 8722447 TI - Any man's death: presidential address. PMID- 8722448 TI - Drug abuse research accomplishments and opportunities: a report from the national perspective. PMID- 8722449 TI - Linking research and service delivery: the unique mission of the Substance Abuse and Mental Health Services Administration. PMID- 8722450 TI - Introduction of Nathan B. Eddy Award recipient. PMID- 8722451 TI - The Nathan B. Eddy Lecture: challenging conventional wisdom about drug abuse. PMID- 8722452 TI - Opioid receptors: molecular and structural studies. PMID- 8722453 TI - The teratogenicity of the drugs of abuse: a symposium. PMID- 8722454 TI - Cannabinoid receptors: pharmacology, second messenger systems and endogenous ligands. PMID- 8722455 TI - Innovative approaches to drug abuse treatment. PMID- 8722456 TI - The role of excitatory amino acids in the actions of abused drugs. PMID- 8722457 TI - Tolerance and sensitization to opioids and cocaine. PMID- 8722458 TI - Overview: antisocial personality and treatment response in substance abusers. PMID- 8722459 TI - Cardiovascular effects of cocaine: underlying mechanisms. PMID- 8722460 TI - Forum on drug research with policy implications. PMID- 8722461 TI - Drugs of abuse and immunosuppression. PMID- 8722462 TI - Hallucinogenic agents: drugs of abuse as neurochemical tools. PMID- 8722463 TI - Behavioral, pharmacologic and neurobiologic variables important to the analysis of drug self-administration: implications for the discovery of potential pharmacotherapies. PMID- 8722464 TI - Dependence studies of new compounds in the rhesus monkey, rat and mouse (1993). PMID- 8722465 TI - Biological evaluation of compounds for their physical dependence potential and abuse liability. XVII. Drug Evaluation Committee of the College on Problems of Drug Dependence, Inc. (1993). PMID- 8722466 TI - Evaluation of new compounds for opioid activity annual report (1993). PMID- 8722467 TI - Progress report from the testing program for stimulant and depressant drugs (1993). PMID- 8722468 TI - Culture and microsurgical manipulation of the early avian embryo. PMID- 8722469 TI - Quail-chick transplantations. PMID- 8722471 TI - Manipulation of the avian segmental plate in vivo. PMID- 8722470 TI - Manipulations of neural crest cells or their migratory pathways. PMID- 8722472 TI - Somite strips: an embryo fillet preparation. PMID- 8722473 TI - Embryo slices. PMID- 8722474 TI - Operations on limb buds of avian embryos. PMID- 8722475 TI - Iontophoretic dye labeling of embryonic cells. PMID- 8722476 TI - Gene transfer using replication-defective retroviral and adenoviral vectors. PMID- 8722477 TI - Manipulating gene expression with replication-competent retroviruses. PMID- 8722478 TI - In situ hybridization analysis of chick embryos in whole mount and tissue sections. PMID- 8722479 TI - Micromass cultures of limb and other mesenchyme. PMID- 8722480 TI - Autonomic and sensory neuron cultures. AB - This chapter has provided a rather detailed protocol for the dissection, dissociation, and culture of autonomic and sensory neurons from the chicken embryo. These protocols are by no means absolute. Many other laboratories that routinely culture these neurons may use techniques that differ significantly from the ones detailed in this chapter. All of the protocols described in this chapter can also be applied to quail embryos, which develop more rapidly but are of comparable size to chicken embryos until about E9. The list of suppliers for the various reagents described in these protocols is also limited. Many other vendors of cell culture products are probably equally reliable. PMID- 8722481 TI - Retinal cultures. PMID- 8722482 TI - Migration and adhesion assays. PMID- 8722483 TI - Cell division and differentiation in avian embryos: techniques for study of early neurogenesis and myogenesis. PMID- 8722484 TI - Time-lapse cinephotomicrography, videography, and videomicrography of the avian blastoderm. PMID- 8722485 TI - Molecules under the microscope. PMID- 8722486 TI - Bioequivalence and generic prescribing: a pharmacy view. PMID- 8722487 TI - Bioequivalence and generic prescribing: an industrial view. AB - The whole concept of bioequivalence is based upon the existence of a clear relationship between drug concentration and clinical effect. To date there are insufficient data available in the form of publications to support this concept. Both the pharmaceutical industry and the regulatory authorities could do more to promote this issue and publish relevant information. The pharmaceutical industry could provide more information on concentration-effect relationships in volunteers and patients. Upon expiry of the patent, regulators could provide estimates of the inter- and intra-subject variability in the pharmacokinetics of a drug in volunteers and patients, assessment of therapeutic windows for drugs and drug classes and their impact on bioequivalence acceptance criteria. Current regulatory guidelines refer to rate and extent of absorption BUT there is no rate parameter which allows products to be compared for both pharmaceutical quality and safety and efficacy. PMID- 8722489 TI - Chemical stability of artesunate injection and proposal for its administration by intravenous infusion. AB - Artesunate, the only artemisinin analogue that can be given intravenously, produces rapid parasite and fever clearance in falciparum malaria. A significant therapeutic problem is a high, late recrudescence rate, probably due to short half-lives of both artesunate and its active metabolite dihydroartemisinin relative to conventional dosing intervals. One method of extending the duration of action of artesunate could be to administer the drug by infusion rather than bolus injection, provided that it is chemically stable at ambient temperature. Artesunate was found to be stable in 0.9% w/v sodium chloride at 9 degrees C, 23 degrees C and 36.5 degrees C for 130, 10.6 and 1.6 h, respectively. Interpolating from an Arrhenius plot, artesunate should be stable for approximately 4 h at 30 degrees C, a temperature representative of ambient conditions in tropical countries. Exposure to light did not affect the degradation rate. Single compartment pharmacokinetic modelling was used to evaluate potential differences in artesunate and dihydroartemisinin plasma concentrations following administration of artesunate by intravenous bolus or infusion. A bolus injection of artesunate at a dose of 4 mg kg-1 gives a peak concentration of 5.3 mg L-1, falling to 0.005 mg L-1 at 5 h. The same dose infused over 4 h results in a peak concentration of 0.92 mg L-1, falling to 0.005 mg L-1 at 8 h. Simultaneous modelling of dihydroartemisinin showed that while its peak plasma concentration was reduced by 27% and the peak delayed by 2.5 h following artesunate administration by infusion, substantially higher concentrations were maintained compared with those predicted after bolus artesunate. These data indicate that artesunate can be administered as a high-dose intravenous infusion, thus avoiding high plasma concentrations. This strategy also has the potential to prolong the duration of antimalarial effect and reduce toxicity, and consequently improve clinical outcome in seriously ill patients. PMID- 8722488 TI - Effects of polyacrylic polymers on the degradation of insulin and peptide drugs by chymotrypsin and trypsin. AB - The purpose of this study was to determine whether carbopol polymers, polyacrylic acid polymers, can inhibit lumenal degradation of insulin, calcitonin and insulin like growth factor I (IGF-I) by trypsin and chymotrypsin and to understand whether reducing the pH of the incubation medium by these polymers results in inhibition. Further, the effects of carbopol polymers on the in-situ absorption of insulin were studied in rats. In saline, carbopol polymers at 1% and 4% (w/v%) inhibited close to 100% of trypsin and chymotrypsin activities against insulin. In 50 mM Tris buffer, carbopol polymers, including 934P, 974P and 971P, at 0.1% only weakly inhibited degradation of calcitonin and insulin by both enzymes; however, as the polymer concentration increased to 0.4%, degradation of insulin, calcitonin, and IGF-I by both enzymes was complete or almost complete. When the Tris buffer was increased to 100 mM, no inhibition was observed at 0.1%. Determination of the final pH of the incubation medium in the presence of polymers revealed that the inhibitory effects of carbopol polymers correlated with the final pH. When the incubation medium has no or low buffer capacity to buffer the protons released by carbopol polymers, these polymers are able to reduce the pH much lower than the optimum pH for the enzyme activities, and thus inhibit proteolytic degradation. When the buffer capacity of the incubation medium increases, the inhibitory effects of carbopol polymers weaken. In-situ absorption of insulin revealed that carbopol polymers improved insulin absorption and induced a significantly greater decline in blood glucose levels. It is concluded that carbopol polymers with strong bioadhesive properties also can inhibit lumenal degradation of peptide hormones, offering multiple advantages for their uses in oral drug delivery. PMID- 8722490 TI - In-vitro evaluation of biphenylyl acetic acid-beta-cyclodextrin conjugates as colon-targeting prodrugs: drug release behaviour in rat biological media. AB - Biphenylyl acetic acid was selectively conjugated to one of the primary hydroxyl groups of beta-cyclodextrin through an ester- or amide-linkage, and the physicochemical properties (aqueous solubility and hydrolysis) were investigated. Aqueous solubility of the conjugates was lower than those of either drug or parent beta-cyclodextrin. The amide conjugate was stable in aqueous solution and in rat biological fluids and gastrointestinal contents. The ester conjugate was hydrolysed to beta-cyclodextrin and biphenylyl acetic acid at moderate rates resulting in a V-shaped rate-pH profile in aqueous solution. The ester conjugate released the drug preferentially when incubated with the contents of caecum or colon, whereas no appreciable drug release was observed on incubation with contents of stomach or intestine, nor on incubation with intestinal or liver homogenates, nor on incubation with rat blood. The present results suggest that the ester-type drug conjugate of beta-cyclodextrin may serve as a colon-targeting prodrug. PMID- 8722491 TI - Prolonged immune response evoked by a single subcutaneous injection of microcapsules having a monophasic antigen release. AB - A model antigen, bovine serum albumin (BSA), was successfully incorporated into microcapsules fabricated from blends of poly(d,l-lactide-co-glycolide) and poly(d,l-lactide). The microcapsules possessed diameters ranging from 10 to 100 microns and exhibited a continuous monophasic release of the protein in-vitro for 3 weeks. They were found to enhance its immunogenicity, thereby potentiating the anti-BSA antibody response following a single subcutaneous injection in mice and rabbits. Enzyme-linked immunosorbent assays demonstrated that the microcapsules provoked high-titre and long-lived immunoglobulin G immune responses over a period of 192 days in mice. When rabbits were immunized by a single subcutaneous injection of BSA-loaded microcapsules, a high level of anti-BSA antibody was still present in the sera obtained at 17 weeks post-injection. The immunization protocol using the BSA-loaded microcapsules was superior to that using BSA dissolved in saline or adsorbed to alum. These microcapsules providing the controlled release of antigens may be valuable in designing better vaccine formulations. PMID- 8722492 TI - The role of gastric mucosal sulphydryls in the ulcer-protecting effects of cisapride. AB - The present study was designed to examine the role of endogenous sulphydryls (SHs) in the gastro-protection induced by cisapride (CIS) (10, 25 and 50 mg kg-1 i.p.), a potent benzamide stimulating gastrointestinal motility in mucosal injury induced by 50% v/v ethanol. Results were compared with those of 5 hydroxytryptamine (5-HT) (10mg kg-1). Ethanol mucosal damage was significantly reduced by treatment with CIS and 5-HT. On the contrary, administration of n ethylmaleimide (NEM) (10 mg kg-1) an SH alkylator, markedly worsened lesion formation and counteracted the protective effect of CIS. Rats pretreated with CIS significantly increased the total sulphydryls as reflected in the non-protein and protein fractions however, 5-HT treatment showed a fall in the non-protein level. The present results suggest that 5-HT-ergic dependent mechanisms have no relation to the gastro-protection afforded by CIS in this experimental model. It is possible that mucosal SHs could be involved. PMID- 8722493 TI - Oral cavity absorption of (R)-8-hydroxy-2-(di-n-propylamino)tetralin and (S)-8 acetyl-2-(di-n-propylamino)tetralin in the rat. AB - The present communication reports on the efficacy of (R)-8-OH-DPAT ((R)-8-hydroxy 2-(di-n-propylamino)tetralin) and (S)-LY-41 ((S)-8-acetyl-2-(di-n propylamino)tetralin) in displaying the 5-HT1A syndrome and decreasing body temperature after administration of the compound subcutaneously into the gastric ventricle or into the oral cavity in the rat. The dose range eliciting a clear cut HT1A syndrome and hypothermia after oral cavity administration was 1/10-1/30 that of the gastric ventricle dose range, but 10-30 times higher than the dose range used for subcutaneous administration of both (R)-8-OH-DPAT and (S)-LY-41. Determination of the concentrations of (R)-8-OH-DPAT in plasma and brain tissue confirmed a higher bioavailability after oral cavity than after gastric ventricle administration; plasma and brain tissue concentrations of the drug were found to be approximately 3 times those after 10 mumol/kg-1 orally than after 100 mumol/kg 1 gastroventrically at 15-60 min after administration of (R)-8-OH-DPAT. These findings suggest that the oral cavity may be an important site for drug delivery of 8-OH-DPAT, LY-41 and other compounds with a low gastrointestinal bioavailability. PMID- 8722494 TI - Effects of 5-lipoxygenase inhibitors on interleukin production by human synovial tissues in organ culture: comparison with interleukin-1-synthesis inhibitors. AB - Prostaglandins and leukotrienes differentially regulate the production of interleukin-1 (IL-1) in monocytes. It was, therefore, decided to investigate the effects of some 5-lipoxygenase inhibitors compared with standard IL-1 synthesis inhibitors on the production of IL-1 by human synovial tissue explants in organ culture. Human synovial (from hip/knee arthroplasty) or porcine tibio-tarsal joint synovial explants were incubated in organ culture in Dulbecco's Modified Eagle's Medium + 5% foetal calf serum in the presence of the test compounds or solvents (controls), or media alone for 1-5 days. Total bioactive IL-1 was assayed in the medium (following serial dilution or with polyethylene glycol 8000 added in some assays to remove inhibitors) using the D-10 T-cell bioassay. Some assays of interleukins 1 alpha, 1 beta, 6 or 8 were performed by ELISA. Of the 5 lipoxygenase inhibitors investigated, MK-886(3-(1-(4-chlorobenzyl)-3-tert-butyl thio-5-isopropylindol-2- yl)-2,2 -dimethyl propanoic acid), L-656,224 ((7-chloro 2-[4-methoxypenyl]methyl)-3 -methyl-5-propyl-4-benzofuranol), PF-5901 and tepoxalin were the most potent inhibitors of IL-1 production. While the PF-5901 was effective at 5-30 microM and tepoxalin was effective at 1-10 microM, the others were the most potent having minimal inhibitory activity in the range of 0.01-0.1 microM. The presumed IL-1-synthesis inhibitors, tenidap and IX-207,887, were inactive at concentrations of 30-50 microM. Leukotriene B4 (1-100 ng/mL-1) added to MK-886 (5 microM)-treated cultures reversed the inhibitory effects of the latter on IL-1, confirming the role of 5-lipoxygenase products in the regulation of IL-1 production. Addition of polyethylene glycol 8000 to MI-886 treated cultures eliminated the inhibitory effects of this drug, suggesting that this drug exerts its effects by promoting production of IL-1 inhibitors. MK-886 also inhibited synovial production of two other pleiotrophic cytokines which it regulates, IL-6 and IL-8. The results suggest that some 5-lipoxygenase inhibitors may be usefully employed in regulating production of those interleukins involved in joint cartilage destruction. PMID- 8722495 TI - Pyridine derivatives stimulate phosphatidylcholine secretion in primary cultures of rat type II pneumocytes. AB - We have examined the effects of pyridine derivatives on phosphatidylcholine secretion in primary cultures of rat type II pneumocytes. Of 12 pyridine derivatives, 4-aminopyridine, 4-dimethylaminopyridine and 4-pyrolidinopyridine had a stimulatory effect on phosphatidylcholine secretion, whereas other derivatives had little effect. The stimulatory effect of 4-aminopyridine was concentration- and time-dependent, and was inhibited by the acetoxymethyl ester of 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (3 microM), an intracellular Ca2+ chelator. In addition, the stimulatory effect of 4 aminopyridine was suppressed by W-7(N-(6-aminohexyl)-5-chloro-1-napthalene sulphonamide)(10 microM), a calmodulin inhibitor, and sphingosine (10 microM) and staurosporine (0-1 microM), protein kinase C inhibitors. These results indicate that several pyridine derivatives stimulate phosphatidylcholine secretion in type II pneumocytes. PMID- 8722496 TI - Pharmacokinetics and brain distribution of magnolol in the rat after intravenous bolus injection. AB - The pharmacokinetics of magnolol in rats was studied after 2, 5, or 10 mg/kg-1 intravenous bolus injection. Plasma concentration-time profiles of magnolol were fitted by a two-compartment open model. There were no significant differences in the elimination half-life, the total body clearance, steady-state volume of distribution, or mean residence time. The area under the plasma-time curve and area under the moment-time curve of magnolol appears to increase proportionally from a dose of 2 to 10 mg/kg-1. These results suggest that magnolol possess linear pharmacokinetics. Notwithstanding, brain concentration of magnolol showed no significant difference among various regions (cerebral cortex, olfactory bulb, hippocampus, striatum, cerebellum, brain stem and rest of brain) after 10 min of magnolol (5 mg/kg-1, i.v.) administration, the mean brain drug concentration was approximately fourfold that of magnolol in plasma. PMID- 8722497 TI - Antimicrobial activity of electrochemical silver ions in nonionic surfactant solutions and in model dispersions. AB - The microbicidal effectiveness against Gram-positive and Gram-negative bacteria and Candida albicans of electrochemical silver ions in aqueous solutions containing nonionic surfactants was investigated. From the perspective of the possible use of anodic silver as a preservative in cosmetic or pharmaceutical preparations, microbicidal efficacy was also studied in oil/water model dispersions. Surfactants and botanical extracts partially inhibited the microbicidal effectiveness of anodic silver. Nevertheless in all the experimental conditions, silver ions reduced the microbial concentration up to 4 log units of the starting inoculum in less than 6 h. The wide microbicidal spectrum and the high rate of kill of silver ions appear, therefore, attractive enough to suggest a possible utilization of anodic silver as a preserving agent. PMID- 8722498 TI - Susceptibility of mice to bacterial and fungal infections after intragastric administration of ebselen. AB - The seleno-organic compound ebselen (2-phenyl-1,2-benzisoselenazol-3(2H)-one) has anti-inflammatory activity and exhibits glutathione peroxidase-like activity in vitro. Ebselen inhibited candidacidal activity over the same range of concentrations as it inhibited the production of microbicidal H2O2 by human neutrophils and macrophage-like cells. Therefore, the long-term administration of ebselen might be expected to induce an immunocompromised state in the host. To examine such a possibility, mice (5-weeks-old ddY, male) were given daily intragastric doses of 0, 10 or 100 mg/kg-1 ebselen for 21 days and then infected intraperitoneally with Candida albicans (10(8) cells/mouse), Pseudomonas aeruginosa (1.5 x 10(7) cells/mouse) or methicillin-resistant Staphylococcus aureus (5 x 10(8) cells/mouse). Ebselen at none of the tested doses affected the increase in body weight of mice during administration of the drug. No evidence was obtained that mice became more susceptible to the various microorganisms after the administration of ebselen at any tested dose. PMID- 8722499 TI - Serum corticosterone increases reflect enhanced uptake inhibitor-induced elevation of extracellular 5-hydroxytryptamine in rat hypothalamus. AB - The increase in extracellular 5-hydroxytryptamine (5-HT) in rat hypothalamus following administration of fluoxetine, a 5-HT-uptake inhibitor, was enhanced by the injection of LY206130(1-[1-H-indol-4-yloxy]-3-[cyclohexylamino]-2-prop ano l maleate), a 5HT1A receptor antagonist, or by L-5-hydroxytryptophan (L-5-HTP), the 5-HT precursor. Elevation of serum corticosterone, measured as a functional output of hypothalamic 5-HT pathways, was greater in rats treated with fluoxetine plus LY206130 or with fluoxetine plus L-5-HTP than in rats treated with the agents alone. Synergism between effects of fluoxetine and L-5HTP has often been reported, but this is the first report of an increased functional effect when a 5 HT1A receptor antagonist is combined with a 5-HT uptake inhibitor to augment the increase in extracellular 5-HT. PMID- 8722500 TI - Acetylcholinesterase inhibition and protection by dizocilpine (MK-801) enantiomers. AB - The optical isomers of the N-methyl-D-aspartate (NMDA) receptor ion-channel blocker dizocilpine (MK-801) were shown to interact with electric eel and rat brain acetylcholinesterase (AChE) in a mixed competitive-noncompetitive way. The (-) form, pharmacologically less active, was the most potent of the two isomers as an AChE inhibitor (Ki for electric eel and rat brain AChE being 6.2 and 17.9 microM, respectively, compared with 200 and 450 microM, respectively, of the (+) form). Both enantiomers premixed with AChE preparations, dose-dependently protected the enzyme from inactivation by diisopropylfluorophosphate (DFP). The maximal protective effects against 40 and 10 microM DFP were in the ranges 10.7 23.8 and 19.5-31.4% of control enzymic activity for the (+) and (-) forms of dizocilpine, respectively. The extent of the protective effect against DFP was increased up to 80.1% of control enzymic activity for (-)-dizocilpine and to 38.4% for (+)-dizocilpine by diluting the enzymic mixtures 1000 times after treatment with the organophosphate agent. The two enantiomers added to AChE 15 min after DFP, failed to reactivate the enzyme. Finally, it was shown that (+)- and (-)-dizocilpine dose-dependently and competitively decreased the DFP bimolecular reaction constant, K(i). We conclude that dizocilpine exerts a protective action towards AChE against irreversible DFP inhibition, but the molecular mechanism of such an action is at present unclear. PMID- 8722501 TI - alpha-Trinositol: a functional (non-receptor) neuropeptide Y antagonist in vasculature. AB - Neuropeptide Y is a sympathetic co-neurotransmitter released with noradrenaline upon sympathetic nerve stimulation. This study describes the ability of a synthetic inositol phosphate, alpha-trinositol(D-myo-inositol 1,2,6-triphosphate; PP 56) to antagonize vasoconstrictor responses to neuropeptide Y in-vitro as well as in-vivo. In human and guinea-pig isolated arteries alpha-trinositol potently (10 nM to 1 microM extracellular concentration) suppressed the constriction evoked by neuropeptide Y alone, the potentiation by neuropeptide Y of noradrenaline-evoked constriction, and the neuropeptide Y-induced inhibition of relaxation. Moreover, in the pithed (areflexive) rat, a non-adrenergic portion of the pressor response to preganglionic sympathetic nerve stimulation was sensitive to alpha-trinositol. As studied in the recently cloned human (vascular-type) Y1 receptor, the action of alpha-trinositol does not occur through antagonism at the neuropeptide Y recognition site nor does it induce allosteric changes of this receptor. However, we found alpha-trinositol to inhibit the rise in intracellular Ca2+ as well as inositol triphosphate concentrations induced by neuropeptide Y. It is, therefore, proposed that alpha-trinositol represents a non-receptor, but yet selective antagonist of neuropeptide Y in vasculature, opening up the possibility to investigate involvement of neuropeptide Y in sympathetic blood pressure control and in cardiovascular disorders. PMID- 8722502 TI - 3-Nitropropionic acid exacerbates [3H]GABA release evoked by glucose deprivation in rat striatal slices. AB - 3-Nitropropionic acid (3-NPA) is a metabolic poison that produces lesions of striatal intrinsic neurones such as gamma-aminobutyric acid (GABA) neurones. This study was carried out to determine whether 3-NPA would impair the ability of striatal GABAergic neurones to withstand hypoglycaemic stress. 3-NPA (500 microM) did not affect [3H]GABA release from striatal slices under normal (11 mM) glucose concentrations. When the glucose concentration was lowered to 0.3 mM, however, 3 NPA greatly potentiated the leakage of [3H]GABA from the slices. Blockage of N methyl-D-aspartate (NMDA) or alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors with 1 microM 5-methyl-10,11-dihydro-5H dibenzo[a,d]cyclohepten-5,10-imine (MK-801) or 10 microM 2,3-dihydroxy-6-nitro-7 sulpha-moylbenzo[F]quinoxaline (NBQX), respectively, or a combination of both, had no effect. However, blockade of voltage-dependent sodium channels with tetrodotoxin totally antagonized the [3H]GABA overflow induced by the combination of 3-NPA and hypoglycaemis. Riluzole (10 to 100 microM), a neuroprotective agent that stabilizes the inactivated state of the voltage-dependent sodium channel, also dose-dependently antagonized the increase in [3H]GABA release induced by the combination of the two stresses. PMID- 8722503 TI - Pharmacological activity of DC-015, a novel potent and selective alpha 1 adrenoceptor antagonist. AB - The pharmacological activity of 3-((4-(2-methoxyphenyl)piperazin-1-yl)methyl)-2,3 dihydroimidaz o(1,2 -c)quinazolin-5(6H)-one (DC-015), a newly synthesized quinazoline derivative, was determined in rat isolated thoracic aorta and pressor responses were determined in spontaneously hypertensive rats (SHR). Experimental results indicated that DC-015 is an alpha 1-adrenoceptor-blocking agent in rat thoracic aorta as revealed by its competitive antagonism of phenylephrine-induced vasocontraction (pA2 = 10.54 +/- 0.55). These effects still persisted in denuded aorta. It was as potent as prazosin (pA2 = 10.04 +/- 0.63). At higher concentration (1.0 microM), DC-015 also expressed 5-hydroxytryptamine (5-HT) receptor competitive antagonism, but this 5-HT blocking effect was not found in the prazosin-administration group. [3H]Inositol monophosphate formation stimulated by phenylephrine (30 microM) in rat thoracic aorta was diminished by DC-015 (3 and 10 nM) and prazosin (10nM); whereas the cAMP content of rat thoracic aorta was not altered by DC-015 and prazosin. Furthermore, intravenous administration of DC-015 and prazosin (both at 0.01, 0.05 and 0.1 mg/kg-1) induced a dose-dependent reduction of mean arterial pressure which reached a maximal effect at 5 mm after injection and persisted over 2 h in SHR. A higher dose of DC-015 (0.1 mg/kg-1, i.v.) did not cause any significant changes in heart rate, whereas, the same dose of prazosin (0.1 mg/kg-1, i.v.) produced a decrease which seems to parallel the time course of the hypotensive response. We can conclude that the DC-015 is a potent, highly selective alpha 1-adrenoceptor antagonist in vascular smooth muscle. PMID- 8722504 TI - Involvement of sigma-receptors in the increase in contraction of mouse vas deferens induced by exogenous ATP. AB - The effects of sigma-receptor ligands on the twitch contraction elicited by the exogenous application of adenosine 5'-triphosphate (ATP) in the unstimulated mouse vas deferens were studied. (-)-Pentazocine, 1,3-di(2-tolyl)guanidine(DTG) and two pairs of optical isomers of 3-(3-hydroxyphenyl)-N-(1-propyl)piperidine(3 PPP) and N-allylnormetazocine (SKF-10,047) potentiated the exogenous application of ATP-induced twitch-type contraction in a concentration-dependent manner, while (+)-pentazocine did not affect it. The order of potentiating ability was: (+)-3 PPP > (-)pentazocine > (-)-SKF-10,047> DTG > (-)-3-PPP > (+)-SKF-10,047. On the other hand, haloperidol and rimcazole, putataive sigma-receptor antagonists, suppressed this twitch contraction. In addition, these antagonists significantly blocked the (+)-3-PPP- and (-)-pentazocine-induced potentiation at concentrations which did not affect contractions per se. These findings indicate that the exogenous application of ATP-induced twitch contraction in the mouse vas deferens is regulated by sigma-receptors. In addition, the present ranking order suggests that the sigma-receptor potentiating the ATP-induced twitch contraction at post junctional sites may differ from the sigma 1- and/or sigma 2-receptor subtypes. PMID- 8722505 TI - Effects of endothelin B antagonist RES-701-1 on endothelin-induced contractile responses in-vivo and in-vitro in guinea-pigs. AB - The effects of an endothelin (ET)-receptor B-specific antagonist, RES-701-1, on ET-induced contraction of guinea-pig trachea and on ET-induced bronchoconstriction in anaesthetized guinea-pigs were investigated. In the epithelium-removed tracheal preparation, 1 x 10(-5) M RES-701-1 inhibited contractions induced by the ETB-specific agonist sarafotoxin S6c (pKB = 6.10). In the epithelium-intact tracheal preparation, RES-701-1 (1 x 10(-5) M)inhibited the ET-3-induced contraction (pKB = 5.27), but enhanced the ET-1-induced contraction significantly and shifted the concentration-response curve to the left. The maximal responses of ET-1- and ET-3-induced contraction were augmented by epithelium removal by 1.5 and 1.8-fold, respectively. Against ET-3-induced contraction in the tracheal preparation without epithelium, RES-701-1 (0.3-10 x 10(-6)M) antagonized the contraction in a concentration-dependent manner (pA2 = 5.9). On the other hand, RES-701-1 (1 x 10(-5)M) did not affect ET-1-evoked responses in the trachea without epithelium. The intravenous administration of ET 1 (1.5 nmol/kg-1) or ET-3 (1.5 nmol/kg-1) evoked a biphasic, fast and sustained bronchoconstriction in anaesthetized guinea-pigs pretreated with propranolol (1.0 mg/kg-1). when administered intravenously, RES-701-1 (0.3 or 1.0 mg/kg-1) showed significant reduction in both phases of bronchoconstriction induced by ET-3. As in the case of ET-1-induced bronchoconstriction, rES-701-1 augmented the sustained phase although a significant reduction of the fast phase was observed. These results indicate that RES-701-1 can inhibit the ET-3 induced airway responses not only in-vitro but also in-vivo. PMID- 8722506 TI - Pharmacological profile of rat pleurisy induced by Bothrops jararaca venom. AB - Bothrops jararaca venom (30 micrograms/site) triggered a marked inflammatory reaction in the pleural cavity that was long-lasting and reproducible. In the first 1 h after pleurisy induction, a significant decrease of total and differential cell count was observed in comparison with control values, despite the gradual enhancement of fluid leakage. A significant increase of cell migration was observed after 3 h of pleurisy induction, due to mononuclear and neutrophil cells that peaked 8 h later and this was followed by a gradual decrease, remaining elevated up to 24 h. In parallel with cell influx, a significant increase of fluid leakage that peaked between 1 and 8 h was observed, being completely abolished after 12 h following pleurisy induction. This inflammatory response was not associated in parallel with significant changes in circulating leucocyte cells and it was significantly inhibited by compound 48/80, cyproheptadine, pyrilamine, dexamethasone, indomethacin and phenidone. Preheating of the venom (100 degrees C) caused a significant decrease of both leakage of fluid and cell migration in the pleural cavity 8 h after pleurisy induction. Previous exposure to the venom (30 micrograms/site, 5 days before) produced a significant decrease of both cell migration and fluid leakage 4 h after triggering pleurisy with the same dose of the venom. Otherwise, prior daily treatment with the venom (10 micrograms/site, 4 days) resulted only in marked fluid leakage reduction 1 h after treating the animals with BJV (30 micrograms/site). These results show that the venom elicits pro-inflammatory effects in the rat pleural cavity which involve the participation of several mediators, including histamine, 5-hydroxytryptamine and products of arachidonic pathways. PMID- 8722507 TI - Antitussive activity of moguisteine enantiomers in guinea-pigs and rats. AB - The antitussive effect of the R-(+)- and S-(-)-enantiomers of moguisteine were evaluated in comparison with the racemate in cough induced by 7.5% citric acid and 30 microM capsaicin aerosol in conscious guinea-pigs. No difference in potency was observed between moguisteine and the enantiomers. The oral ED50 values (with 95% confidence limits) for moguisteine, R-(+)- and S-(-)-enantiomers were respectively: 20.4 (12.9-26.6), 20.9 (14.9-26) and 21.6 (11.8-30.0) mg/kg-1 in cough provoked by citric acid and 17.7 (12.5-29.8), 18.9 (14.1-30.1) and 20.5 (15.1-36.6) mg/kg-1 in cough induced by capsaicin. The acute oral and intraperitoneal toxicities of the enantiomers and moguisteine in the rat are very similar. These findings suggest that the use of either enantiomer does not offer any advantage over the racemate. PMID- 8722508 TI - The tricyclic antidepressants clomipramine and citalopram induce apoptosis in cultured human lymphocytes. PMID- 8722509 TI - Morphology of incipient mesoderm formation in the rabbit embryo: a light- and retrospective electron-microscopic study. AB - Mesoderm formation is a hallmark of vertebrate gastrulation and, at the same time, one of the prime examples for epithelio-mesenchymal transformation. Recent advances in experimental embryology and molecular biology have clarified the role of growth factors and genes in this process; however, its microscopic anatomy in higher vertebrates is still far from clear. Therefore, the present study describes the morphology of mesoderm formation in the rabbit embryo, a species which may be representative for both the avian and the mammalian embryo in this respect. Serial semithin sections were correlated with topographical landmarks in surface views of embryonic discs at 6.4, 6.5, and 6.6 days post conceptionem, and selected semithin sections were reembedded for ultrastructural analysis. Mesoderm cells are shown to be generated by ingression of bottle-shaped epiblast cells in the area of the posterior node and the primitive streak. Here, basal endocytotic pits and absence or discontinuity of the basal lamina are taken as suggestive evidence for specific removal of extracellular matrix material. Within the bottle shaped cells most organelles are concentrated in a narrow apical neck which will subsequently constitute the 'trailing end' of the ingressing mesoderm cells. These features support the assumption that most principles of epithelio mesenchymal transformation seen during primary mesenchyme formation in the sea urchin also apply to mesoderm formation in vertebrates. However, transient tripartite zonula adherens-type junctions are formed apically between ingressing mesoderm cells and the neighboring epiblast cells. They are interpreted here as being responsible for maintaining supracellular integrity of the embryonic disc during the shedding of mesoderm cells in the amniote embryo. PMID- 8722510 TI - A subset of SBA lectin-binding proteins isolated from myocardial-conditioned media transforms cardiac endothelium into mesenchyme. AB - It has been shown that the inductively active proteins for cardiac mesenchyme formation are localized to a particulate form of extracellular matrix that resembles adheron-like complexes. These complexes are extractable from the embryonic heart using EDTA and can be visualized with the lectin SBA (Glycine max). In addition, the growth medium obtained from embryonic myocardial cell cultures has also been shown to support mesenchyme formation. However, except for the identification of EDTA extract and conditioned media, all previous experiments analyzing this system have relied on negative type results (i.e. the loss of biological activity) to show a relationship between the particulate matrix and the transformation process. We report here that SBA affinity chromatography can be used to partially purify a subset of proteins from myocardial conditioned medium which elicits the transformation of endothelial cells into mesenchyme. In addition, a polyclonal antibody made against this subset of proteins is specific for the in situ particulate matrix and recognizes several proteins in conditioned medium and EDTA extracts. This antibody is also specific for matrix particulates in other areas of the embryo that undergo an epithelial/mesenchymal interaction. These results provide the most direct evidence to date that conditioned medium is equivalent to the hypothesized inductively active particulate matrix. In addition, the data provides evidence that conditioned medium can be used to identify the functional role of the components of the particulate matrix in mesenchyme formation. PMID- 8722511 TI - Immunofluorescent analysis of the spinal cord in dysraphic mice. AB - Development of the spinal cord was analyzed immunocytochemically at 10-12 days of gestation in normal and dysraphic embryos of the loop-tail mutant mouse, using an anti-neural cell adhesion molecule (NCAM) and the lectin concanavalin A (Con A) as histological markers for evaluating neural cell organization and distribution. The normal and abnormal embryos showed similar patterns of reactivity to anti NCAM and Con A, even though the neural folds were open and everted in the abnormal embryos, with displacement of the dorsal root ganglia. In the abnormal embryos the floor plate was similar to that in normal embryos, as evidenced by its increased anti-NCAM and Con A labeling relative to that in the rest of the neuroepithelium. Moreover, each lateral end of the everted abnormal neuroepithelium developed an attenuated 'roof plate' that appeared to be structurally similar to the normal roof plate. However, some of these 'roof plates' exhibited prominent clusters of labeled and nonlabeled cells, especially in the 10-day embryos. In addition, whereas normal embryos showed strong luminal labeling of the neuroepithelial cells with Con A, comparable regions in the abnormals were spotty and poorly defined except for the 'roof plate' and floor plate. The results indicate that dorsoventral polarity in the spinal cord, as assessed structurally and histochemically, develops essentially normally in abnormal dysraphic embryos, even though the topographic relationships of the abnormal neural tube are disturbed. PMID- 8722512 TI - An anatomical and embryological study of the clavicle in cats (Felis domestus) and sheep (Ovis aries) during the prenatal period. AB - The prenatal development of the clavicular area was studied in two species: the domesticated sheep, which lacks a clavicle, and the cat, with a nonfunctional, rudimentary clavicle. A morphological and computerized morphometric study of the clavicle was performed in 18 cat embryos between 25 and 48 days of gestation, and in 12 sheep embryos of 37-45 days. One group of embryos was processed with double staining in toto according to Hanken and Wassersug in 1981. The other group was examined by histological techniques: hematoxylineosin-Alcian blue and picrosirius. In both species, clavicular ossification is delayed (27% of gestation time elapsed in sheep and 53% in cats) compared to 16% in humans. Histological and morphological differences in shape and length of the clavicle were observed in both species. The clavicle is transient in sheep, whereas in the cat it persists with little change. In neither species does secondary cartilage develop. In cats, the periosteum is well developed with active osteoblasts, whereas in sheep the clavicle is surrounded by a single layer of epitheloid cells and the periosteum is less developed and contains osteoclasts. These results suggest that the morphogenetic pattern for the clavicle is altered at about day 34 in cats and day 40 in sheep, and is subsequently partly inhibited in the former and blocked in the latter. PMID- 8722513 TI - Stereological analysis of the urethra in sexually intact and spayed female dogs. AB - The perfusion-fixed urethrae of 5 sexually intact nulliparous and 5 ovariohysterectomized beagles were analyzed stereologically. Different urethral tissue components were estimated stereologically in each of the three proximal urethral quarters and compared between the two groups of dogs. Considerable individual variability within all groups was observed. In all the three urethral quarters considered, average smooth muscle and connective tissue volumes were generally lower in spayed than in intact animals, both in absolute and relative terms. However, only the difference between the mean relative volumes of urethral quarter I was statistically significant. In contrast, mean absolute and relative volumes of the vascular plexus were generally greater in the ovariohysterectomized dogs as compared to the animals of the other group- deviations in statistical relevance concerning the first urethral quarter were noted. Mean epithelial and striated muscle volumes did not vary essentially between the spayed and intact group of dogs. Despite the fact that all findings were not always statistically meaningful, our research nevertheless demonstrates that ovariohysterectomy decreases smooth muscle mass and connective tissue; it also elevates vascular plexus volumes in the urethra of spayed animals. Moreover, our results imply a general weakening of the urethral wall as a sequel to ovariohysterectomy, and this, in turn, could be seen to affect the closure mechanism of the urethra. PMID- 8722514 TI - Electron-microscopic study of gamma-aminobutyric acid immunoreactivity in the chief cells of the mouse carotid body. AB - The present study shows gamma-aminobutyric acid (GABA) immunoreactivity in almost all chief cells of the mouse carotid body. GABA immunoreaction products were distributed diffusely in the cytoplasm as well as in the dense-cored granules. These findings suggest that GABA may be localized in the cytoplasm and granules, and may be released from the cells. PMID- 8722515 TI - Adaptations of diaphragm neuromuscular junction following inactivity. AB - We hypothesized that differences exist in the morphological adaptations of neuromuscular junctions (NMJs) on different fiber types in response to prolonged inactivation. Two weeks of inactivity of both phrenic motoneurons and diaphragm muscle was induced by spinal cord hemitransection at C2 (spinal isolation; SI). A three-color fluorescent immunocytochemical technique, combined with laser scanning confocal microscopy, was used to create two- (2D) and three-dimensional (3D) images of NMJs and obtain morphological information concerning: (1) innervating axons and presynaptic nerve terminals; (2) motor endplates (postsynaptic apparatus consisting of acetylcholine receptors), and (3) myosin heavy chain (MHC) phenotype of muscle fibers. In both sham controls (CTL) and SI animals, planar (2D) and surface (3D) areas of motor endplates and nerve terminals on type II muscle fibers (anti-fast MHC immunoreactive) were smaller than on type I (nonimmunoreactive to anti-fast MHC) fibers, when normalized for fiber diameter. The number of branches, total branch length and perimeter of both motor endplates and nerve terminals were greater for NMJs on type II fibers than on type I fibers. The extent of overlap between nerve terminal and endplate was greater on type I fibers than on type II fibers. After SI, there was a significant expansion of NMJs on type II fibers. Planar and surface areas of motor endplates and nerve terminals, number of endplate and nerve terminal branches, total branch length, and perimeter were all increased on type II fibers following SI. The extent of overlap of nerve terminal and endplate increased on type II fibers, approaching that observed in type I fiber NMJs. These results indicate that neuromuscular inactivation leads to a selective expansion of type II fiber NMJs through addition of new terminal area, and elongation of existing terminal branches. These changes may represent a compensatory effort to improve neuromuscular transmission. PMID- 8722516 TI - Locational relationship of the supraorbital notch or foramen and infraorbital and mental foramina in Koreans. AB - The morphology and locational relationship of the supraorbital notch/foramen, infraorbital foramen, and mental foramen were studied from photographs of 124 Korean skulls (male 35, female 18, unknown sex 71). The infraorbital foramen was on the sagittal plane passing through the supraorbital notch/foramen (36.4%), or lateral to the plane (63.6%). The mental foramen was either on the plane (69.3%), lateral to the plane (21.8%), or medial to the plane (9.0%). The supraorbital notch/foramen, infraorbital foramen, and mental foramen were on the same sagittal plane in 38.1% of the cases. The supraorbital notch (69.9%) was found more frequently than the supraorbital foramen (28.9%). The average distance from the median plane to the center of the supraorbital notch/foramen, infraorbital foramen, and mental foramen was 22.7, 27.2 and 24.4 mm, respectively. The average length of the line from the center of the supraorbital notch/foramen to the center of the infraorbital foramen was 45.6 mm, and the angle of this line to the sagittal plane was 5.8 degrees. The average distance from the infraorbital margin to the center of the infraorbital foramen was 8.6 mm, and that from the inferior margin of mandible to the center of the mental foramen was 15.5 mm in males and 14.0 mm in females. The average horizontal width of the supraorbital notch/foramen, infraorbital foramen, and mental foramen was 4.7, 4.8 and 2.4 mm, respectively. Most commonly, the infraorbital foramen was lateral to the sagittal plane of the supraorbital notch/foramen, and the mental foramen was on that plane. This locational relationship would be helpful clinically to determine the location of the infraorbital and mental foramina, by palpation of the supraorbital notch. PMID- 8722517 TI - One hundred years of anatomical terminology. PMID- 8722518 TI - Too much compassion? PMID- 8722519 TI - U.K. moves toward compulsory vaccination. PMID- 8722520 TI - Public goods and fair prices. Balancing technological innovation with social well being. PMID- 8722521 TI - The run on Ritalin. Attention deficit disorder and stimulant treatment in the 1990s. PMID- 8722522 TI - "Give me children or I shall die!" New reproductive technologies and harm to children. PMID- 8722523 TI - The value of a uterus. PMID- 8722524 TI - Reconceiving the family. The process of consent in medical decisionmaking. PMID- 8722525 TI - Outcomes research and practice guidelines. Upstream issues for downstream users. PMID- 8722526 TI - Effects of speech and language disorders on raters' perceptions. AB - Using semantic differential scales with nine trait pairs, 465 students at two universities rated five descriptions, one portraying an individual without a disorder and four depicting communicative disorders. Statistical analyses indicated that the descriptions with no disorder and with a language disorder were rated significantly lower in ambition than those depicting stuttering, voice, or articulation disorders. Other differences emerged when the raters were divided by geographical location and age. Location was a significant factor in the responses for the traits of social adjustment, employability, and ambition. Age of respondent correlated significantly with ratings of tension and employability. PMID- 8722527 TI - Young children's acquisition of the location aspect of American Sign Language signs: parental report findings. AB - The acquisition of the location aspect of American Sign Language signs was examined in 9 young children of deaf parents. In monthly home visits, the parents demonstrated on videotape how their children formed each newly-acquired sign in their lexicons; these videotaped records served as the basis for the present analyses. Sign locations, overall, were produced with relatively high accuracy: 83.5% were correct on average across the different signs in the children's early lexicons. Certain sign locations were found to be more easily or readily acquired than others. Highly contrasting locations (forehead, chin, on and in front of the trunk) were acquired first. Among the locations typically acquired later were those that involved complex handshapes, provided a small area for a point of contact, or required the active signing hand to cross the body's midline. The location aspect was interpreted as playing a central role in young children's early sign language acquisition. PMID- 8722528 TI - Metalinguistic and pragmatic abilities of participants in adult literacy programs. AB - The relationship between oral and written language skills was explored in this study involving 97 participants enrolled in an adult literacy program. Pragmatic language skills were assessed in videotaped conversational dyads which were later analyzed for level of conversational assertiveness and responsiveness and for the status of basic interactional skills. Those behaviors which were judged as being most likely to detract from communicative effectiveness were body movement, eye gaze, and facial expression. Metalinguistic/ semantic language skills were assessed with the Test of Word Knowledge (TOWK) (Wiig & Secord, 1991). A significant relationship was found between total scores on the TOWK and reading levels of subjects as they entered the program. Post-testing was conducted approximately 6 to 8 months after the initial testing. These results were available for 22 of the subjects and indicated that, even for adults, some aspects of oral language improve as reading levels increase. PMID- 8722529 TI - Predictive factors of persistence and recovery: pathways of childhood stuttering. AB - This article presents broad preliminary findings from a longitudinal study of stuttering pertaining to differentiation of developmental paths of childhood stuttering, as well as possible early prediction of High Risk, Low Risk, and No Risk for chronic stuttering. More than 100 preschool children who stutter have been closely followed for several years from near the onset of stuttering using a multiple data collection system, with 45 nonstuttering children serving as controls. Thirty-two stuttering and 32 control subjects who have progressed through several stages of the investigation were identified for the present indepth analyses. They represent four subgroups: I. Persistent Stuttering; II. Late Recovery; III. Early Recovery; IV. Control. Comparative data for the groups with special reference to differences in frequency of disfluency, acoustic features, phonologic skills, language development, nonverbal skills, and genetics are presented. The results suggest several promising predictors of recovery and chronicity. PMID- 8722530 TI - Antimicrobial resistance and Helicobacter pylori. PMID- 8722531 TI - A meta-analysis of studies on the safety and efficacy of aminoglycosides given either once daily or as divided doses. AB - We performed a meta-analysis of randomised clinical studies in which the efficacy and toxicity of the same total daily dose of aminoglycosides administered once daily was compared with multiple divided daily dosing for treating human infections. Of twenty-eight publications identified from a literature search using Medline 19 publications of 20 study comparisons involving 2881 patients met the criteria for analysis. Netilmicin was investigated in 11 studies, amikacin in seven studies and gentamicin in two studies but no studies of tobramycin met the inclusion criteria. The meta-analysis showed that there was a small, statistically significant difference in clinical efficacy of 3.5% (95% confidence intervals 0.5% to 6.5%, P = 0.027) in favour of once-daily administration but no significant differences in bacteriological efficacy or nephrotoxicity were detected. Auditory and vestibular toxicity rates were low for all agents and no differences in these toxicities were identified between once-daily or multiple dose administration regimens either clinically or by audiometry or electronystagmography. Aminoglycosides can be given once-daily without loss of efficacy or increased toxicity offering greater simplicity and potentially improved cost-effectiveness than can be achieved by giving these drugs in divided doses. PMID- 8722532 TI - Temperature-dependent aminoglycoside resistance in Stenotrophomonas (Xanthomonas) maltophilia; alterations in protein and lipopolysaccharide with growth temperature. AB - Clinical strains of Stenotrophomonas (Xanthomonas) maltophilia often show large, growth temperature-dependent, variations in their susceptibility (TDVS) to aminoglycoside antibiotics. Strains showing more than a fourfold increase in susceptibility between 30 degrees and 37 degrees C (TDVS+ strains; n = 23) were contrasted with those showing lesser variation (TDVS- strains; n = 15) in studies of growth temperature-dependent variation in protein and cell-wall lipopolysaccharide (LPS) electrophoresis patterns in an attempt to determine the mechanism of TDVS. Several proteins showed increased intensity with increasing growth temperature. These comprised bands at c. 65, 55, 42.5, 26 and 21.5 kDa in the whole cell proteins, an outer membrane protein band at c. 21.5 kDa, and cytoplasmic membrane protein bands at c. 42.5 and 27 kDa. Two whole cell protein bands at c. 30 and 24 kDa and three outer membrane protein bands at c. 45, 30 and 24 kDa decreased in intensity with increasing growth temperature. However, there was no correlation with the extent of variation in susceptibility, either in the extent of temperature dependent changes in protein banding patterns, or the presence or absence of specific protein bands. By contrast, temperature-dependent variation in LPS patterns correlated well with TDVS. TDVS+ strains yielded intense ladder patterns of more than 30 discrete bands, and the mean molecular weight of the ladder pattern was markedly higher at growth temperatures < or = 30 degrees C, than at > or = 37 degrees C. TDVS- strains gave a clearly distinct high mol. wt LPS banding pattern showing fewer, less intense bands and a smaller and less consistent shift in mean molecular weight with temperature. Strains which were clearly resistant at 30 degrees and 37 degrees C, had a high mol. wt. polysaccharide component but an absence of the typical LPS-ladder pattern. We conclude that the temperature-dependent variation in the aminoglycoside susceptibility of this species was not correlated with any detectable change in protein composition, but correlated well with changes in LPS structure. PMID- 8722533 TI - A reverse transcriptase-PCR based assay for in-vitro antibiotic susceptibility testing of Chlamydia pneumoniae. AB - Infections caused by Chlamydia spp are an important cause of human disease, and the accuracy and reproducibility of antimicrobial susceptibility tests for these bacteria could have considerable clinical implications. We have developed a reverse transcriptase PCR (RT-PCR) based method to determine the antibiotic susceptibility of Chlamydia spp., and compared this with conventional tests using immunofluoresence (IF) staining. The MICs of antimicrobial agents for a test strain of Chlamydia pneumoniae were higher by RT-PCR as compared with IF staining, indicating the greater stringency of the former method. Using RT-PCR, doxycycline and tetracycline were the most active agents (MIC 1 mg/L), followed by erythromycin (1.6 mg/L), and ciprofloxacin (16 mg/L). Neither trimethoprim nor sulphamethoxazole (400 mg/L) inhibited growth as assessed by both techniques. The RT-PCR based method may thus represent an improved and less time consuming assay for in-vitro determination of the antibiotic susceptibility of Chlamydia spp. PMID- 8722534 TI - Use of the coagulase gene typing method for detection of carriers of methicillin resistant Staphylococcus aureus. AB - Strains of Staphylococcus aureus can be typed on the basis of the polymorphism of the coagulase gene. DNA fragments generated after amplification by polymerase chain reaction (PCR) of the variable region of this gene and digested with the restriction enzyme HaeIII can be compared by their restriction fragment length polymorphism (RFLP). Seventy-nine of 86 (91.8%) methicillin-resistant S. aureus (MRSA) strains isolated in various hospitals had a characteristic RFLP pattern. This pattern differed from those of 32 methicillin-susceptible S. aureus (MSSA). Only one MSSA shared with MRSA the same RFLP pattern. After modification, we applied this method to the rapid detection of MRSA from 255 nasal swabs from patients hospitalized in intensive care units. When screened by plating on Chapman agar, 55 of these samples contained MRSA, whereas 40/55 yielded the expected restriction profile after amplification by PCR. No DNA was amplified by PCR in 9/55 samples and restriction profiles were uninterpretable in six cases. When compared to culture, the positive and negative predictive values of the PCR test were 100% and 93%, respectively. The specificity was 100% and the sensitivity was 72.7%. Since control of spread of MRSA strains in hospitals is based in part on rapid isolation of carriers, this method which allows detection of epidemic MRSA in nasal swabs within a day could be helpful, though culture would still be necessary to confirm the result. PMID- 8722535 TI - In-vitro evaluation of beta-lactamase inhibition by latamoxef and imipenem. AB - The in-vitro 50% inhibitory concentrations (IC50s) of latamoxef and imipenem against a set of plasmid-mediated beta-lactamases including TEM-1, TEM-3, TEM-5 and TEM-10 were determined by an enzymatic method using nitrocefin as substrate. The IC50s of both antibiotics against extended-spectrum beta-lactamases were below 0.2 mg/L. The conventional spectrum beta-lactamase TEM-1 was not inhibited by either antibiotic at the highest concentration tested. Except for TEM-10 for which the IC50s of the two antibiotics were the same, imipenem showed significantly greater activity than latamoxef against TEM-3 and TEM-5. Clavulanic acid taken as a control demonstrated greater and wider inhibitory activity, but on the other hand it has no significant antibiotic activity. PMID- 8722536 TI - A pharmacodynamic evaluation of ciprofloxacin and ofloxacin against two strains of Pseudomonas aeruginosa. AB - The greater potency of ciprofloxacin in vitro to that of ofloxacin against Pseudomonas aeruginosa may be potentially offset by the more favorable pharmacokinetic profile of the latter drug. In order to test this hypothesis, we generated time concentration kill curves for P. aeruginosa ATCC 27853 and a clinical isolate P. aeruginosa PSA 9258 using an in-vitro model to simulate the pharmacokinetic characteristics found in vivo for ciprofloxacin at a peak concentration (CPmax) of 5 mg/L and an elimination T1/2 of 4.5 h, and ofloxacin at a CPmax of 5 mg/L and a T1/2 of both 4.5 h and 6 h, and at a CPmax of 8.0 mg/L and a T1/2 of 6 h. A 3 log10 kill (T3 kill) of P. aeruginosa ATCC 27853 was achieved in 0.15 h by ciprofloxacin and of P. aeruginosa PSA 9258 in 0.09 h. Ofloxacin at a CPmax of 8 mg/L and T1/2 of 6 h achieved a T3 kill of P. aeruginosa ATCC 27853 in 0.74 h and of P. aeruginosa PSA 9258 in 0.16 h. The area under the kill curve (AUKC) was 1.10 x 10(4)and 1.96 x 10(3) mL-h/cfu for ciprofloxacin against P. aeruginosa ATCC 27853 and P. aeruginosa PSA 9258, respectively and that of ofloxacin at CPmax 8 mg/L and a T1/2 of 6 h was 9.78 x 10(4)and 2.20 x 10(4) mL-h/cfu respectively. Significant differences (P > or = 0.05) were evident between ciprofloxacin and all ofloxacin regimens against P. aeruginosa ATCC 27853 but not against P. aeruginosa PSA 9258. There was a poor correlation (r = 0.22) between the AUKC and area under the time concentration curve (AUC) for P. aeruginosa ATCC 27853 but a strong correlation (r = 0.96) between the AUKC and area under the inhibitory curve (AUIC). Similar results were obtained for P. aeruginosa PSA 9258 for which the correlation between AUKC and AUC was weak (r = 0.10) whereas that between the AUKC and AUIC was strong (r = 0.93). When the data for both P. aeruginosa were combined, a correlation coefficient of r = 0.04 for AUC and r = 0.80 for AUIC was found. These limited data suggest that fluoroquinolones can be compared using the AUIC for specific bacterial isolates. In addition, the larger AUC, higher CP, and longer T1/2 of ofloxacin in vivo did not fully compensate for the intrinsic differences in the antibiotic susceptibility against P. aeruginosa. PMID- 8722537 TI - In-vitro and in-vivo antibacterial activity of LB10517, a novel catechol substituted cephalosporin with a broad antibacterial spectrum. AB - LB10517 is a new injectable cephalosporin with a broad spectrum of antibacterial activity against Gram-positive and Gram-negative bacteria. LB10517 inhibited 90% of methicillin-susceptible Staphylococcus aureus (MSSA) at 0.25 mg/L (MIC90), and was 8-fold more active than cefpirome. LB10517 was two- or four-fold more active than cefpirome against methicillin-susceptible Staphylococcus epidermidis (MSSE), Streptococcus pyogenes and Enterococcus faecalis. Both methicillin-resistant S. aureus (MRSA) and methicillin-resistant S. epidermidis (MRSE) were highly resistant to all compounds. LB10517 activity against most Enterobacteriaceae was comparable to or greater than that of cefpirome, and it also showed high activity against Pseudomonas aeruginosa. In a mouse septicaemia model, LB10517 exhibited excellent protective effects. In a respiratory tract infection model, the protective effect of LB10517 was comparable to that of cefpirome and ceftazidime. It was highly stable to hydrolysis by beta-lactamases, showing better physiological efficiency for TEM-9 than the other test compounds. LB10517 had the most potent antibacterial activity against beta-lactamase producing resistant strains. LB10517 did not induce beta-lactamase production in Enterobacter cloacae 1194E. PMID- 8722538 TI - Unorthodox antibiotic combinations including ciprofloxacin against high-level gentamicin resistant enterococci. AB - Development of high-level gentamicin resistance among enterococci represents a serious therapeutic problem as it precludes synergy between aminoglycosides and cell-wall active agents. As part of a search for active antibiotic combinations against enterococci with high-level gentamicin resistance, we tested by the time kill curve method the efficacy of ciprofloxacin combined with ampicillin, trimethoprim-sulphamethoxazole, vancomycin or teicoplanin against ten isolates of Enterococcus faecium, three of Enterococcus casseliflavus and 13 of Enterococcus faecalis that exhibited a MIC of gentamicin > or = 2000 mg/L. Most of the E. faecium were also resistant to ampicillin and to ciprofloxacin. The combination of ciprofloxacin with ampicillin was bactericidal against five of seven E. faecium strains that exhibited a ciprofloxacin MIC < or = 4 mg/L, but was inactive against the three E. faecium that were highly resistant to ciprofloxacin. This combination was also bactericidal against the E. casseliflavus and all the E. faecalis strains. The combination of ciprofloxacin with trimethoprim-sulphamethoxazole was bactericidal against five of the seven E. faecium and seven of the nine E. faecalis strains with a ciprofloxacin MIC < or = 4 mg/L. No bactericidal activity of this combination was seen against the enterococci that were highly resistant to either ciprofloxacin or to trimethoprim sulphamethoxazole. The combination of ciprofloxacin with glycopeptides was inactive against E. faecium and E. casseliflavus and against E. faecalis, it was either ineffective or antagonistic; in only one case it was bactericidal. Five strains of E. faecium were resistant to all antibiotic combinations tested. PMID- 8722539 TI - The incidence and epidemiology of beta-lactam resistance in Haemophilus influenzae. AB - Consecutive isolates of Haemophilus influenzae were collected by the Public Health Laboratory, Bath between 1 June 1992 and 31 May 1993. Of 379 apparently distinct isolates, 216 originated from the respiratory tract, 102 from eyes and 61 from other sites. The minimum inhibitory concentrations of amoxycillin, amoxycillin/clavulanate, cefaclor, cefuroxime, cefotaxime and cefpodoxime were determined for each isolate. Forty strains (10.6%) were beta-lactamase producers. MIC50 and MIC90 values and the range of MICs were determined for all isolates. The overall resistance rates were: amoxycillin (MIC > 1.0 mg/L), 22.7%; amoxycillin/clavulanate (MIC > 1.0 mg/L), 14.8%; cefuroxime (MIC > 1.0 mg/L), 18.5%, (MIC > 4.0 mg/L), 5.5%; cefaclor (MIC > 8 mg/L), 15.6%; cefpodoxime (MIC > 1.0 mg/L), 0.3%; cefotaxime (MIC > 1.0 mg/L), 0%. Twenty non-beta-lactamase producing but beta-lactam resistant strains (cefuroxime MIC > 4.0 mg/L) were matched with 20 susceptible strains on the basis of patient age, sex, and specimen type. The strains were characterised by outer-membrane protein (OMP), random amplified polymorphic DNA (RAPD) and ribotyping patterns. Eleven of the 20 resistant strains were indistinguishable by the methods used, suggesting spread of a single beta-lactam resistant, non-beta-lactamase producing clone. The distribution of resistant strains within the local community was plotted geographically. PMID- 8722540 TI - Ciprofloxacin resistant Campylobacter spp. in humans: an epidemiological and laboratory study. AB - From the end of April 1991 until the end of 1991, 2209 isolates of Campylobacter spp. have been collected in Plymouth PHL of which 91 (4.1%) were resistant to ciprofloxacin. None of the 91 patients involved had taken a quinolone, but 30/91 (33%) had travelled abroad (16 to the Iberian peninsula) in the three months preceding isolation of the ciprofloxacin-resistant Campylobacter spp. In the case control study 12/15 (80%) of the cases had recently consumed poultry as had 20/24 (83%) of controls with enteritis due to ciprofloxacin-susceptible Campylobacter spp. A small study of poultry purchased from the supermarket revealed that only 1/37 campylobacters isolated from 64 UK bred chickens was resistant to ciprofloxacin, whereas 7/26 campylobacters isolated from 50 imported chickens were ciprofloxacin-resistant. Of the 75 clinical isolates of ciprofloxacin resistant Campylobacter spp. subjected to detailed analysis, 68 were Campylobacter jejuni, six were Campylobacter lari, and one was Campylobacter coli. All isolates from man and poultry were resistant to ciprofloxacin, norfloxacin, sparfloxacin and tosufloxacin, and there was an association between fluoroquinolone-resistance and increased MICs of tetracycline. The range of susceptibility to erythromycin and kanamycin were typical of the species. gyrA from C. jejuni P6 (a case with history of travel to Spain) and C. jejuni P16 (isolate from imported chicken) contained point mutations corresponding to an amino acid substitution of isoleucine for threonine at codon 86. It has been suggested that veterinary use of quinolones, notably enrofloxacin, is providing a selective pressure for emergence of resistance to ciprofloxacin amongst human isolates. Now that enrofloxacin has been licensed for use in broiler flocks in the UK, it will be interesting to monitor the prevalence of resistance of campylobacters to quinolones in UK-produced poultry and in UK-acquired human infection. PMID- 8722541 TI - Characterization of antiviral activity of a sesquiterpene, triptofordin C-2. AB - The activities of 13 sesquiterpenes isolated from Tripterygium wilfordii Hook fil. var. regelii Makino were studied against herpes simplex virus type 1 (HSV-1) in vitro. Among these compounds, only triptofordin C-2 showed a selectivity index of more than 10. The compound, which could also inhibit the replication of human cytomegalovirus (HCMV), did not affect either adsorption or penetration of HSV-1 to host cells, but showed moderate virucidal activity against several enveloped viruses including HSV-1, HCMV, measles virus and influenza A virus. Triptofordin C-2 suppressed viral protein synthesis of infected cells when added at early steps of HSV-1 replication and exerted inhibition of translation of the transcripts of the immediate early genes. When acyclovir and triptofordin C-2 were evaluated in combination for antiviral activity against HSV-1 replication, additive antiviral effects were observed for this virus. PMID- 8722542 TI - Doxycycline or ciprofloxacin prophylaxis and therapy against experimental Yersinia pestis infection in mice. AB - The efficacy of doxycycline and ciprofloxacin against an experimental plague infection was assessed by comparing the median lethal dose (MLD) of Yersinia pestis in antibiotic-treated and untreated mice. The MLD of Y. pestis GB strain in untreated mice by the intra-peritoneal route was 23 cfu. If ciprofloxacin dosage (20 or 40 mg/kg twice daily) was initiated 48 h before infection, it afforded complete protection against an intra-peritoneal challenge of 5.24 x 10(7) cfu. Ciprofloxacin therapy initiated 24 h post-challenge was less protective, the MLD was raised to 2.0 x 10(5) and 2.2 x 10(5) cfu for 40 and 20 mg/kg respectively. Doxycycline dosage (40 mg/kg twice daily) initiated 48 h prior to infection raised the MLD to 1.6 x 10(4) cfu, but other prophylactic and therapeutic regimes were ineffective against challenges greater than 6.76 x 10(2) cfu. Ciprofloxacin may therefore be a useful antibiotic to consider for the treatment of plague. PMID- 8722543 TI - Clinical pharmacokinetics of meropenem after the first and tenth intramuscular administration. AB - We investigated the pharmacokinetics of meropenem after the first and tenth i.m. administration in patients with respiratory tract infections. Ten patients (mean age 63.8 +/- 5.2 years) received meropenem 500 mg tds for at least ten doses, and plasma and urine antibiotic concentrations were determined by microbiological assay. After the first injection a mean peak plasma concentration of 7.93 +/- 1.29 mg/L was observed at 1 h. Trough levels at 8 h (0.29 +/- 0.16 mg/L) were detectable in five of ten treated patients. The mean terminal half-life was 1.08 +/- 0.2 h with an area under the curve (AUC) value of 23.8 +/- 4.59 mg/L.h, and a cumulative urinary recovery at 8 h of 48.43 +/- 3.12%. There was no evidence of change in the pharmacokinetics of meropenem after repeated i.m. administration, though the mean peak plasma concentration and AUC value were slightly increased. The accumulation ratio (assessed using AUC values) was 1.18 +/- 0.19 after multiple doses and was considered to be of little kinetic and clinical importance. Moreover, many of the trough concentrations of meropenem were below the limit of detection of the assay. After i.m. administration meropenem concentrations exceeded 0.5 mg/L for longer than previously described following i.v. infusion. No adverse events were reported. PMID- 8722544 TI - Elimination of bacteraemia after dental extraction: comparison of erythromycin and clindamycin for prophylaxis of infective endocarditis. AB - Erythromycin and clindamycin are currently recommended for antibiotic prophylaxis of infective endocarditis in predisposed patients allergic to penicillin undergoing oral invasive procedures. Thirty-eight healthy patients were randomized to receive either erythromycin (1 g) or clindamycin (0.6 g) orally 1.5 h prior to dental extraction. Blood samples for microbiological investigation were collected before, during and 10 min after surgery and were processed by lysis filtration under anaerobic conditions. The incidence of bacteraemia with viridans streptococci was 79% in the erythromycin group and 74% in the clindamycin group. No statistically significant difference was noted in incidence or magnitude of bacteraemia with viridans streptococci or anaerobic bacteria between the two groups, at any sampling time. Ninety-six aerobic and 133 anaerobic strains recovered from the blood samples were tested for their susceptibility to erythromycin and clindamycin as well as to penicillin V and ampicillin. The antimicrobials were found to be highly active against the majority of bacteria except for some enterococci, staphylococci and veillonella. Protection from endocarditis by prophylaxis with erythromycin or clindamycin must be due to elimination of bacteria at a later stage in the development of the disease, rather than by elimination of bacteria from blood during the short period of postoperative bacteraemia. PMID- 8722545 TI - Detection of mutations conferring extended-spectrum activity on SHV beta lactamases using polymerase chain reaction single strand conformational polymorphism (PCR-SSCP). AB - Single strand conformational polymorphism (SSCP) is a recently developed technique used to detect single base mutations in short PCR-generated amplimers. The method has been adapted and applied to differentiation of beta-lactamase genes. Each of the five standard SHV strains used produced a unique SSCP pattern, allowing the possibility of rapid identification of the SHV genes of other isolates. A clinical isolate that phenotypically produced SHV-5 yielded a pattern of major bands indistinguishable from that of the SHV-5 standard strain, illustrating the applicability of this technique. We therefore report a reliable and reproducible technique that can be applied to the characterisation of the SHV beta-lactamases. PMID- 8722546 TI - Comparative in-vitro activity of CP-99219, a new quinolone, against respiratory pathogens. AB - The comparative in-vitro activity of CP-99219, a new quinolone, against Haemophilus influenzae (150 isolates), Moraxella catarrhalis (100), Streptococcus pneumoniae (80) and Group A beta-haemolytic streptococci (40) was determined using an agar dilution technique. CP-99219 was the most active compound tested against M. catarrhalis (MIC50 = 0.015 mg/L, MIC90 = 0.03 mg/L). Ceftriaxone, CP 99219 and ciprofloxacin were the three most active agents tested against H.influenzae. CP-99219 showed good activity, 16-fold greater than that of ciprofloxacin, against S.pneumoniae (MIC50 = 0.12 mg/L; MIC90 = 0.25 mg/L) and was also active against Group A streptococci. Clinical studies regarding the use of CP-99219 in respiratory tract infections seem indicated. PMID- 8722547 TI - Comparative distribution of resistance patterns and serotypes in Pseudomonas aeruginosa isolates from intensive care units and other wards. AB - The resistance patterns and O-serotypes of 2952 Pseudomonas aeruginosa isolates were studied and the relationship between patterns and serotypes was investigated. The penicillinase-producing and cephalosporinase-overproducing phenotypes were significantly more frequent in intensive care units than other wards, but there was no difference for the intrinsic resistance phenotype. The predominant serotypes were O6, O11 and O1. The incidence of serotype O12 was low. O11 isolates were more common in intensive care units and more resistant to all antibiotics than other isolates. Most O12 isolates had a penicillinase-producing phenotype and were resistant to aminoglycosides and ciprofloxacin, but susceptible to fosfomycin. PMID- 8722548 TI - In-vitro antifungal activity of sertaconazole, bifonazole, ketoconazole, and miconazole against yeasts of the Candida genus. AB - The in-vitro antifungal activity of sertaconazole against 110 strains of Candida yeasts (50 Candida albicans, 15 Candida glabrata, 2 Candida guilliermodii, 8 Candida krusei, 1 Candida kefyr, 8 Candida parapsilosis and 26 Candida tropicalis) was assessed in comparison with bifonazole, ketoconazole, econazole and miconazole. The majority of the strains were clinical isolates; some reference strains were included. A commercial agar diffusion method (NeoSensitabs, Rosco, Taastrup, Denmark) in Shadomy's modified medium pH 7 was used. Using the manufacturer's criteria, 86.4% of the strains were classified as "sensitive" to sertaconazole. The only strain classified as "resistant" to sertaconazole was the control reference strain of C. albicans. The remaining strains were classified as "moderately sensitive". The sensitivity/resistance percentages for the other antifungals tested were 75.5/1.8 for ketoconazole, 71.8/2.7 for miconazole, 63.7/13.6 for econazole, and 59.1/5.5 for bifonazole. Sertaconazole showed a higher antifungal activity than that of the other antimycotics, tested in vitro which was statistically significant (P < 0.001), as well as a lower resistance rate than that of econazole, bifonazole and ketoconazole. PMID- 8722549 TI - Concentrations of cefpodoxime in plasma, adenoid, and tonsillar tissue after repeated administrations of cefpodoxime proxetil in children. AB - Cefpodoxime proxetil was administered to 36 children undergoing tonsillectomy, adenoidectomy or both. It was very well tolerated. The detectable tissue concentrations of cefpodoxime were moderate but remained constant (approximately 0.05 mg/kg) 3, 6, and 12 h after the last dose of the drug, while the respective plasma concentrations were declining. This suggests the possibility of twice daily administration. However, 30% of children did not have quantifiable concentrations in the tonsil and more than half the adenoids did not have quantifiable levels. Whether a higher dosage would lead to higher and more satisfactory tissue concentrations is a matter for further investigation. PMID- 8722550 TI - Zidovudine absorption and small intestinal function in HIV seropositive patients. AB - Zidovudine absorption was evaluated in HIV seropositive patients with (n = 15) and without diarrhoea (n = 20) in a standardised prospective pharmacokinetic study using single oral 200 mg doses. Zidovudine was rapidly absorbed with large peak variation (Cmax: 1.10 +/- 0.43 mg/L). There were no significant associations between pharmacokinetic parameters and presence of diarrhoea, CD4 counts, red blood cell folate concentrations, stool cultures/leucocytes or the lactulose/mannitol absorption test. Mean diarrhoea AUC (mg/L.h) was 1.13 +/- 0.30 and 1.07 +/- 0.36 in non-diarrhoeal patients. Antidiarrhoeals increased AUC and Cmax values in a small, non statistically significant subset of diarrhoea patients. Although zidovudine absorption varies significantly, our data do not support dosage individualisation based on any of the above parameters. PMID- 8722551 TI - Peripheral sensory disturbances related to treatment with fluoroquinolones. AB - The symptoms and possible risk factors of peripheral sensory disturbances related to fluoroquinolones are reviewed on the basis of 37 reports submitted to the Swedish Adverse Drug Reactions Advisory Committee. In 25 patients (68%), symptoms occurred within 1 week after start of treatment. Paraesthesia was the most common complaint and occurred in 81% of the cases. Fifty-one per cent of the reports concerned numbness/hypoaesthesia, 27% pain/hyperaesthesia and 11% muscle weakness. Seventy-one per cent of the patients recovered within 2 weeks after drug discontinuation. Possible predisposing factors were impaired renal function, diabetes, lymphatic malignancy and treatment with another drug known to cause neuropathy. PMID- 8722552 TI - The first molecular characterization of tetracycline-resistant Neisseria gonorrhoeae from Uruguay. PMID- 8722553 TI - Drug protonation and pH in relation to the lethal action of tamoxifen on Candida albicans. PMID- 8722554 TI - A simple micro-method for time-kill studies amenable to routine laboratory use. PMID- 8722555 TI - Reassessment of methods for testing the susceptibility of Haemophilus influenzae to clarithromycin. PMID- 8722556 TI - In-vitro activity of trovafloxacin (CP99,219) tested by two methods against 150 vancomycin-resistant enterococcal isolates. PMID- 8722557 TI - A comparative trial of short term therapy with omeprazole plus either amoxycillin or azithromycin for Helicobacter pylori eradication. PMID- 8722558 TI - The long pore gets molecular. PMID- 8722559 TI - Unidirectional K+ fluxes through recombinant Shaker potassium channels expressed in single Xenopus oocytes. AB - We describe a method to evaluate the ratio of ionic fluxes through recombinant channels expressed in a single Xenopus oocyte. A potassium channel encoded by the Drosophila Shaker gene tested by this method exhibited flux ratios far from those expected for independent ion movement. At a fixed extracellular concentration of 25 mM K+, this channel showed single-file diffusion with an Ussing flux-ratio exponent, n', of 3.4 at a membrane potential of -30 mV. There was an apparent, small voltage dependence of this parameter with n' values of 2.4 at -15 and -5 mV. These results indicate that the pore in these channels can simultaneously accommodate at least four K+ ions. If each of these K+ ions is in contact with two water molecules, the minimum length of the pore is 24 A. PMID- 8722560 TI - Ca2+ currents in cerebral artery smooth muscle cells of rat at physiological Ca2+ concentrations. AB - Single Ca2+ channel and whole cell currents were measured in smooth muscle cells dissociated from resistance-sized (100-microns diameter) rat cerebral arteries. We sought to quantify the magnitude of Ca2+ channel currents and activity under the putative physiological conditions of these cells: 2 mM [Ca2+]o, steady depolarizations to potentials between -50 and -20 mV, and (where possible) without extrinsic channel agonists. Single Ca2+ channel conductance was measured over a broad range of Ca2+ concentrations (0.5-80 mM). The saturating conductance ranged from 1.5 pS at 0.5 mM to 7.8 pS at 80 mM, with a value of 3.5 pS at 2 mM Ca (unitary currents of 0.18 pA at -40 mV). Both single channel and whole cell Ca2+ currents were measured during pulses and at steady holding potentials. Ca2+ channel open probability and the lower limit for the total number of channels per cell were estimated by dividing the whole-cell Ca2+ currents by the single channel current. We estimate that an average cell has at least 5,000 functional channels with open probabilities of 3.4 x 10(-4) and 2 x 10(-3) at -40 and -20 mV, respectively. An average of 1-10 (-40 mV and -20 mV, respectively) Ca2+ channels are thus open at physiological potentials, carrying approximately 0.5 pA steady Ca2+ current at -30 mV. We also observed a very slow reduction in open probability during steady test potentials when compared with peak pulse responses. This 4-10-fold reduction in activity could not be accounted for by the channel's normal inactivation at our recording potentials between -50 and -20 mV, implying that an additional slow inactivation process may be important in regulating Ca2+ channel activity during steady depolarization. PMID- 8722561 TI - Ion permeation and block of M-type and delayed rectifier potassium channels. Whole-cell recordings from bullfrog sympathetic neurons. AB - Ion permeation and conduction were studied using whole-cell recordings of the M current (I(M)) and delayed rectifier (IDR), two K+ currents that differ greatly in kinetics and modulation. Currents were recorded from isolated bullfrog sympathetic neurons with 88 mM [K+]i and various external cations. Selectivity for extracellular monovalent cations was assessed from permeability ratios calculated from reversal potentials and from chord conductances for inward current. PRb/PK was near 1.0 for both channels, and GRb/GK was 0.87 +/- 0.01 for IDR but only 0.35 +/- 0.01 for I(M) (15 mM [Rb+]o or [K+]o). The permeability sequences were generally similar for I(M) and IDR: K+ approximately Rb+ > NH4+ > Cs+, with no measurable permeability to Li+ or CH3NH3+. However, Na+ carried detectable inward current for IDR but not I(M). Nao+ also blocked inward K+ current for IDR (but not IM), at an apparent electrical distance (delta) approximately 0.4, with extrapolated dissociation constant (KD) approximately 1 M at 0 mV. Much of the instantaneous rectification of IDR in physiologic ionic conditions resulted from block by Nao+. Extracellular Cs+ carried detectable inward current for both channel types, and blocked I(M) with higher affinity (KD = 97 mM at 0 mV for I(M), KD) approximately 0.2 M at 0 mV for IDR), with delta approximately 0.9 for both. IDR showed several characteristics reflecting a multi ion pore, including a small anomalous mole fraction effect for PRb/PK, concentration-dependent GRb/GK, and concentration-dependent apparent KD's and delta's for block by Nao+ and Cso+. I(M) showed no clear evidence of multi-ion pore behavior. For I(M), a two-barrier one-site model could describe permeation of K+ and Rb+ and block by Cso+, whereas for IDR even a three-barrier, two-site model was not fully adequate. PMID- 8722562 TI - Mechanism of action of K channel openers on skeletal muscle KATP channels. Interactions with nucleotides and protons. AB - The molecular mechanisms underlying the actions of K channel openers (KCOs) on KATP channels were studied with the patch clamp technique in excised inside-out patches from frog skeletal muscle fibers. Benzopyran KCOs (levcromakalim and SR 47063) opened channels partially blocked by ATP, ADP, or ATP gamma s, with and without Mg2+, but they had no effects in the absence of internal nucleotides, even after channel activity had significantly declined because of rundown. The effects of KCOs could therefore be attributed solely to a competitive interaction between KCOs and nucleotides, as confirmed by observations that ATP decreased the apparent affinity for KCOs and that, conversely, KCOs decreased ATP or ADP sensitivity. Protons antagonized the action of the non-benzopyran KCOs, pinacidil and aprikalim, by enhancing their dissociation rate. This effect resembled the effect of acidification on benzopyran KCOs (Forestier, C., Y. Depresle, and M. Vivaudou. FEBS Lett. 325:276-280, 1993), suggesting that, in spite of their structural diversity, KCOs could act through the same binding sites. Detailed analysis of the inhibitory effects of protons on channel activity induced by levcromakalim or SR 47063 revealed that, in the presence of 100 microM ATP, this effect developed steeply between pH 7 and 6 and was half maximal at pH 6.6. These results are in quantitative agreement with an allosteric model of the KATP channel possessing four protonation sites, two nucleotidic sites accessible preferentially to Mg(2+)-free nucleotides, and one benzopyran KCO site. The structural implications of this model are discussed. PMID- 8722563 TI - Osmotic gradient-induced water permeation across the sarcolemma of rabbit ventricular myocytes. AB - The mechanism of water permeation across the sarcolemma was characterized by examining the kinetics and temperature dependence of osmotic swelling and shrinkage of rabbit ventricular myocytes. The magnitude of swelling and the kinetics of swelling and shrinkage were temperature dependent, but the magnitude of shrinkage was very similar at 6 degrees, 22 degrees, and 37 degrees C. Membrane hydraulic conductivity, Lp, was approximately 1.2 x 10(-10) liter.N-1.s 1 at 22 degrees C, corresponding to an osmotic permeability coefficient, Pf, of 16 microns.s-1, and was independent of the direction of water flux, the magnitude of the imposed osmotic gradient (35-165 mosm/liter), and the initial cell volume. This value of Lp represents an upper limit because the membrane was assumed to be a smooth surface. Based on capacitive membrane area, Lp was 0.7 to 0.9 x 10(-10) liter.N-1.s-1. Nevertheless, estimates of Lp in ventricle are 15 to 25 times lower than those in human erythrocytes and are in the range of values reported for protein-free lipid bilayers and biological membranes without functioning water channels (aquaporin). Evaluation of the effect of unstirred layers showed that in the worst case they decrease Lp by < or = 2.3%. Analysis of the temperature dependence of Lp indicated that its apparent Arrhenius activation energy, Ea', was 11.7 +/- 0.9 kcal/mol between 6 degrees and 22 degrees C and 9.2 +/- 0.9 kcal/mol between 22 degrees and 37 degrees C. These values are significantly greater than that typically found for water flow through water filled pores, approximately 4 kcal/mol, and are in the range reported for artificial and natural membranes without functioning water channels. Taken together, these data strongly argue that the vast majority of osmotic water flux in ventricular myocytes penetrates the lipid bilayer itself rather than passing through water-filled pores. PMID- 8722564 TI - Kinetic isoforms of intramembrane charge in intact amphibian striated muscle. AB - The effects of the ryanodine receptor (RyR) antagonists ryanodine and daunorubicin on the kinetic and steady-state properties of intramembrane charge were investigated in intact voltage-clamped frog skeletal muscle fibers under conditions that minimized time-dependent ionic currents. A hypothesis that RyR gating is allosterically coupled to configurational changes in dihydropyridine receptors (DHPRs) would predict that such interactions are reciprocal and that RyR modification should influence intramembrane charge. Both agents indeed modified the time course of charging transients at 100-200-microM concentrations. They independently abolished the delayed charging phases shown by q gamma currents, even in fibers held at fully polarized, -90-mV holding potentials; such waveforms are especially prominent in extracellular solutions containing gluconate. Charge movements consistently became exponential decays to stable baselines in the absence of intervening inward or other time-dependent currents. The steady-state charge transfers nevertheless remained equal through the ON and the OFF parts of test voltage steps. The charge-voltage function, Q(VT), shifted by approximately +10 mV, particularly through those test potentials at which delayed q gamma currents normally took place but retained steepness factors (k approximately 8.0 to 10.6 mV) that indicated persistent, steeply voltage dependent q gamma contributions. Furthermore, both RyR antagonists preserved the total charge, and its variation with holding potential, Qmax (VH), which also retained similarly high voltage sensitivities (k approximately 7.0 to 9.0 mV). RyR antagonists also preserved the separate identities of q gamma and q beta species, whether defined by their steady-state voltage dependence or inactivation or pharmacological properties. Thus, tetracaine (2 mM) reduced the available steady-state charge movement and gave shallow Q(VT) (k approximately 14 to 16 mV) and Qmax (VH) (k approximately 14 to 17 mV) curves characteristic of q beta charge. These features persisted with exposure to test agent. Finally, q gamma charge movements showed steep voltage dependences with both activation (k approximately 4.0 to 6.5 mV) and inactivation characteristics (k approximately 4.3 to 6.6 mV) distinct from those shown by the remaining q beta charge, whether isolated through differential tetracaine sensitivities, or the full approximation of charge-voltage data to the sum of two Boltzmann distributions. RyR modification thus specifically alters q gamma kinetics while preserving the separate identities of steady-state q beta and q gamma charge. These findings permit a mechanism by which transverse tubular voltage provides the primary driving force for configurational changes in DHPRs, which might produce q gamma charge movement. However, they attribute its kinetic complexities to the reciprocal allosteric coupling by which DHPR voltage sensors and RyR-Ca2+ release channels might interact even though these receptors reside in electrically distinct membranes. RyR modification then would still permit tubular voltage change to drive net q gamma charge transfer but would transform its complex waveforms into simple exponential decays. PMID- 8722565 TI - Pharmacology of the skate electroretinogram indicates independent ON and OFF bipolar cell pathways. AB - Organization of afferent information into parallel ON and OFF pathways is a critical feature of the vertebrate visual system. All afferent visual information in the vertebrate retina reaches the inner plexiform layer (IPL) via bipolar cells. It is at the bipolar cell level that separation of ON and OFF information first appears for afferent information from cones. This may also hold true for the rod pathway of cold-blooded vertebrates, but not for mammals. The all-rod retina of the skate presents an opportunity to examine such pathways in a retina having but a single class of photoreceptor. Immunocytochemical evidence suggests that both ON and OFF bipolar cells are present in the skate retina. We examined the pharmacology of the skate electroretinogram (ERG) to test the hypothesis that independent ON and OFF bipolar cell pathways are functional as rod afferent pathways from outer to inner plexiform layer in the skate. 100 microM 2-amino-4 phosphonobutyric acid (APB) reversibly blocked the skate ERG b-wave. A small d wave-like OFF component of the ERG revealed by DC recording of response to a prolonged (10 s) flash of light was reduced or blocked by 5 mM kynurenic acid (KYN). We found that addition of 200 microM picrotoxin to the Ringer's solution revealed prominent ON and OFF components of the skate ERG while reducing the c wave. These ON and OFF components were reversibly blocked by 100 microM APB and 5 mM KYN, respectively. Reversible block of the OFF component by KYN was also accomplished in the presence of 500 microM N-methyl-DL-aspartate. From these findings, we conclude that ON and OFF bipolar cells are likely to be functional as parallel afferent interplexiform pathways in the all-rod retina of the skate. PMID- 8722566 TI - Distribution and characterization of functional amiloride-sensitive sodium channels in rat tongue. AB - The role of amiloride-sensitive Na+ channels (ASSCs) in the transduction of salty taste stimuli in rat fungiform taste buds has been well established. Evidence for the involvement of ASSCs in salt transduction in circumvallate and foliate taste buds is, at best, contradictory. In an attempt to resolve this apparent controversy, we have begun to look for functional ASSCs in taste buds isolated from fungiform, foliate, and circumvallate papillae of male Sprague-Dawley rats. By use of a combination of whole-cell and nystatin-perforated patch-clamp recording, cells within the taste bud that exhibited voltage-dependent currents, reflective of taste receptor cells (TRCs), were subsequently tested for amiloride sensitivity. TRCs were held at -70 mV, and steady-state current and input resistance were monitored during superfusion of Na(+)-free saline and salines containing amiloride (0.1 microM to 1 mM). Greater than 90% of all TRCs from each of the papillae responded to Na+ replacement with a decrease in current and an increase in input resistance, reflective of a reduction in electrogenic Na+ movement into the cell. ASSCs were found in two thirds of fungiform and in one third of foliate TRCs, whereas none of the circumvallate TRCs was amiloride sensitive. These findings indicate that the mechanism for Na+ influx differs among taste bud types. All amiloride-sensitive currents had apparent inhibition constants in the submicromolar range. These results agree with afferent nerve recordings and raise the possibility that the extensive labeling of the ASSC protein and mRNA in the circumvallate papillae may reflect a pool of nonfunctional channels or a pool of channels that lacks sensitivity to amiloride. PMID- 8722567 TI - Characterization of cDNA encoding for phosphoglucose isomerase of rice (Oryza sativa L.). AB - Two types of genes (Pgi-a and Pgi-b) encoding phosphoglucose isomerase (PGI; EC 5.3.1.9) were cloned from cDNA libraries of rice cultured cells (Oryza sativa L.). Pgi-a and Pgi-b consisted of 2132 and 2030 nucleotides, respectively. The homology between these genes was 93.0% at nucleotide level. The homology scores between these genes in protein coding region and 3' non-coding region were 95.6% and 79.4%, respectively. PGI proteins encoded by Pgi-a and Pgi-b consisted of 567 and 568 amino acid, respectively, sharing 95.8% homology at amino acid sequences. Of 11 PGI genes from other plant species and organisms whose amino acid sequences had been determined, a dicotyledonous plant Clarkia lewisii PGI showed the highest homology (about 80%) with rice PGIs. GC contents at the third position of rice PGI genes were about 40%. In order to confirm the enzyme activity of the protein encoded by the rice cDNA, Pgi-a was subcloned into an expression vector, pBluescript II SKp, which was introduced into Escherichia coli. The transformant had an additional PGI activity from Pgi-a. PMID- 8722568 TI - Identification of three highly expressed replacement histone H3 genes of alfalfa. AB - One genomic and six cDNA clones for the replacement histone H3.2 protein of alfalfa (Medicago sativa) were isolated and sequenced. By gene organization they represent 3 distinct genes. PCR methods were used to confirm that only three intron-bearing histone H3.2 genes of this type exist per haploid genome. They co exist with approximately 56 copies of the previously characterized replication dependent, intronless histone H3.1 variant gene. Comparison of the relative expression of few constitutive H3.2 genes with the high S phase expression of the abundant cell cycle-dependent H3.1 genes by mRNA levels and protein synthesis measurements revealed that the replacement histone H3.2 genes are very highly expressed. Structural analysis of the genomic replacement H3.2 gene revealed a unique feature. A repeated polypyrimidine sequence motif in the 5' untranslated region of this gene replaces the ubiquitous intron present in all known replacement H3 genes. A hypothesis is presented that this motif and other, non randomly distributed polypyrimidine sequences in the introns of replacement histone H3 genes of alfalfa and Arabidopsis, may affect nucleosome assembly. Chromatin repression of these replacement genes would be avoided, consistent with the high, constitutive expression of replacement H3 histone genes in plants. PMID- 8722569 TI - Cloning and nucleotide sequence of a xylanase-encoding gene from Streptomyces sp. strain EC3. AB - Using the p1J702 vector, a xylanase-encoding gene (xin) of Streptomyces sp. EC3 has been cloned by functional complementation of a mutant of Streptomyces lividans TK24, producing xylanase at a very low level. Normal level of xylanase synthesis was restored in at least three clones, containing the same 3802 bp Sstl DNA fragment. In this fragment, several open reading frames (ORFs) have been identified, one of which coded for a xylanase; the products of the other ORFs did not show homology with any of the already known proteins. The complete nucleotide sequence of the 3802 bp Ssti insert has been determined on both strands. Xylanase is very probably synthesized as a 240 amino acid (aa) precursor (25949 Da) including a long (49 aa) signal sequence presenting significant similarity with the signal sequences of other Streptomyces xylanase genes. The xylanase aa sequence showed a clear homology with the aa sequences of other xylanases of the glycanase G family. The xln gene has been introduced into Streptomyces parvulus, a naturally xylanase-negative species. In contrast with its expression in Streptomyces sp. EC3, in S. parvulus, xln was expressed constitutively, a probable consequence of the absence of a regulatory system. PMID- 8722570 TI - Identification and mapping of tRNA genes on the Helianthus annuus mitochondrial genome. AB - The physical map for seventeen tRNA genes on the mitochondrial genome of the dicotyledonous plant Helianthus annuus has been established. Eleven are genuine mitochondrial genes, while the other six show a high degree of similarity with the chloroplast counterparts. The genes, with the exception of the genuine trnS(GCT) and of the chloroplast-like trnV and trnP, are expressed. The comparison of the organization of some tRNA genes in the H. annuus mitochondrial genome with that of similar genes detectable in other plants reveals that their association is common to several dicotyledons. PMID- 8722571 TI - Sequence of the highly conserved gene encoding the human 54kDa subunit of signal recognition particle. AB - The complete sequence of the human gene encoding the 54kDa subunit of the signal recognition particle has been isolated from a cDNA library. Degenerate oligonucleotides based on the 5' and 3' coding region of the canine gene and a mammalian codon usage table, were used to amplify the sequence using PCR. The nucleotide sequence of the human gene shows that the human sequence shares a 95.8% nucleotide sequence homology and 100% amino acid sequence homology to it's canine counterpart. The sequence has been given the accession number X86373 in the EMBL database. PMID- 8722572 TI - Significant divergence in nucleotide sequences for beta-tubulin from different laboratory strains of Chinese hamster ovary cells. AB - The nucleotide sequences for isotype 1 beta-tubulin cDNAs cloned from different laboratory strains of Chinese hamster ovary (CHO) cells were compared and found to contain an unexpected number of sequence differences in both translated and untranslated regions of the gene. The results indicate significant changes in the DNA, but not protein, sequence while the cells have been in culture and reveal sequences in the 5' and 3' untranslated regions that have resisted these changes. PMID- 8722573 TI - Nucleotide sequence of the internal transcribed spacers and 5.8S region of ribosomal DNA in Pinus pinea L. AB - The nucleotide sequence of the first internal transcribed spacer (ITS1) belonging to different ribosomal RNA genes from Pinus pinea are reported. The analyzed ITS1 can be distinguished on the basis of their length, being one 2631 bp and the other 271 bp long. Nucleotide comparison of these regions did not show appreciable sequence homology. The larger ITS1 contains five tandem arranged subrepeats with size ranging between 219 bp and 237 bp. The nucleotide sequence of the 5.8S and the ITS2 regions belonging to the larger ribosomal RNA gene are also reported. PMID- 8722574 TI - Cloning and sequencing of the dnaK locus in Streptomyces coelicolor A3(2). AB - The dnaK operon of Streptomyces coelicolor A3(2) was cloned by the DNA-probing method using synthetic oligonucleotides designed on the basis of two of the most conserved regions in 30 different DnaK proteins (HSP70). The isolated insert-a BamHI 5.6-kb fragment-was sequenced and shown to contain three open-reading frames organized in an operon and coding for proteins analogous to DnaK, GrpE and DnaJ, successively. PMID- 8722575 TI - Nucleotide sequence of exons 5 to 9 of the p53 tumour-suppressor gene of the horse (Equus caballus). AB - An evolutionary conserved 1.3 kb fragment corresponding to the horse p53 tumour suppressor gene was PCR amplified, cloned and the nucleotide sequence determined. The p53 fragment encoded exons 5 to 9 and the intervening introns. The nucleotide sequence and the predicted aminoacid sequence showed a high level of homology with human and donkey p53 sequences. PMID- 8722576 TI - Identification of thyroid blocking antibodies and receptor epitopes in autoimmune hypothyroidism by affinity purification using synthetic TSH receptor peptides. AB - To examine the interaction of immunoglobulins from patients with newly diagnosed hypothyroidism with the TSH receptor (TSHr), we tested protein-A purified IgG in an ELISA assay with a series of peptides representing the entire extracellular domain (ECD) of human TSHr. Antibodies bound, on average, 4.1 peptides (range 0 16) per patient, and antibodies from 26 of 30 patients (86.6%) demonstrated binding to at least one peptide. Six of the 20-mer peptides (61, 151, 181, 301, 361, 376) were most frequently recognized. These were used to construct affinity columns and separate IgGs from 10 patients into bound and unbound fractions. All fractions were tested for their ability to stimulate and inhibit cAMP generation in FRTL-5 cells. Inhibitory IgGs were purified from 9 patients (90%), suggesting that the incidence of blocking antibodies (TBAb) in autoimmune hypothyroidism is higher than previously reported. 7 of 10 patients had antibodies that recognized peptide 361 further supporting the importance of this epitope in TBAb binding. Anti-microsomal and anti-thyroglobulin antibodies did not co-purify with inhibitory antibodies, and were always in the unbound fractions. We found no correlation between the pattern of antibody binding or bioactivity with clinical manifestations of hypothyroidism. CONCLUSIONS: (1) The majority of patients with autoimmune hypothyroidism have antibodies against the TSHr-ECD that recognized linear epitopes. Most have antibodies directed at more than one site and the pattern is quite heterogeneous. (2) Six sites (noted above) are most frequently recognized. (3) Inhibitory antibodies are distinct from anti-microsomal and anti thyroglobulin antibodies. PMID- 8722577 TI - Naturally occurring anti-idiotypic antibodies to anti-phosphotyrosine in systemic lupus erythematosus interact with SRC-homology 2 domains. AB - We have recently identified in SLE sera naturally occurring anti-idiotypic antibodies against anti-phosphotyrosine antibodies. Analysis of immunochemical properties of these anti-idiotypic antibodies suggest that they are of beta/gamma type mimicking the antigen. The interaction between these anti-idiotypes and SH2 domains of various fusion proteins was analysed by immunoprecipitation and immunoblotting. Our data demonstrate that these anti-idiotypic antibodies specifically bind SH2 domains, with the highest affinity for SH2 domain of lck protein tyrosine kinase. The significance of this interaction is discussed. PMID- 8722578 TI - Amino acid sequence of a tryptic peptide of human thyroglobulin reactive with sera of patients with thyroid diseases. AB - Autoantibodies to human thyroglobulin (hTg) are found in the sera of many patients with thyroid diseases. To localize epitopes recognized by these autoantibodies, hTg was incubated with tryspin for 4 hours at 37 degrees C under non-reducing conditions. Releasing peptides from hTg in their natural conformation. These peptides were then analyzed by western immunoblot using either autoantibodies from patients with autoimmune thyroiditis or murine monoclonal antibodies (mAb) produced against hTg. The autoantibodies reacted primarily with two low molecular weight peptides with apparent molecular weights (MWap) of 15 and 20 kDa. The pattern of tryptic peptides recognized by these autoantibodies resembled that of one of the mAbs (137C1), as shown by immunoblots in either one or two dimensional SDS-PAGE. To characterize these peptides further, they were separated by a high performance liquid chromatography (HPLC). The column separated the 4-hour tryptic digest of hTg into multiple peptide peaks. Further analysis by SDS-PAGE showed that one of these peaks contained the 15 kDa peptide. The 15 amino acid sequence at the amino-terminus of this peptide was determined. This amino acid sequence (KVPTFATPWPDFVPR) corresponds to a unique sequence near the carboxyl-terminal end of hTg, starting with amino acid 2657. The size of the peptide indicates that it extends to the carboxyl-terminal end of hTg. This fragment contains one of the antigenic sites of hTg that binds autoantibodies from patients with autoimmune thyroid disease. PMID- 8722579 TI - Immunological mimicry between retinal S-antigen and group A streptococcal M proteins. AB - Immunological mimicry between host and microbial proteins has been suggested as a potential mechanism in the development of uveitis in humans. In this study immunological crossreactivity between anti-streptococcal monoclonal antibodies (MAbs) and the human eye was investigated. In indirect immunofluorescence, we demonstrated novel immunological crossreactivity of two anti-streptococcal MAbs (27 and 112) with the rod outer (and inner) segments of the retina of the human eye. In further studies, retinal S-Ag, a uveitogenic protein in the rod outer (and inner) segments, was found to react with the anti-streptococcal MAbs. In addition, several uveitogenic peptides of S-Ag were recognized by the anti streptococcal MAbs. In the ELISA and Western immunoblot, anti-S-Ag MAbs crossreacted with group A streptococci and the streptococcal M protein further demonstrating sites of antigenic similarity. Homology between the retinal S-Ag and streptococcal M protein was observed in amino acid sequences repeated in the B repeat region of the streptococcal M5 protein. These data show that retinal S antigen has immunological similarities with streptococcal M protein, a major virulence determinant and strong bacterial cell surface antigen. PMID- 8722580 TI - Agalactosyl IgG and antibody specificity in rheumatoid arthritis, tuberculosis, systemic lupus erythematosus and myasthenia gravis. AB - Agalactosyl IgG (Gal(0) is a glycoform lacking terminal galactose from the oligosaccharides situated on the Fc. The percentage of circulating IgG that is Gal(0) is increased in a a number of autoimmune diseases, and in certain chronic infections associated with autoantibody production. However it is not known whether this represents decreased galactosylation of all IgG, or an increase in the relative concentration of a subset of agalactosyl antibodies of specificity relevant to the disease process. Since there is currently no way to separate agalactosyl from galactosylated IgG, we devised an assay for the relative degree of galactosylation of antibody to tetanus toxoid (TT), an antigen irrelevant to the diseases studied, and compared this value with the %Gal(0) of the whole circulating IgG. In rheumatoid arthritis (RA) and tuberculosis (TB), a raised %Gal(0) in serum IgG was reflected in a parallel rise in the extent to which antibody to TT was agalactosyl. In SLE a rise in %Gal(0) was seen in the presence of very little rise in agalactosyl anti-TT, and in myasthenia gravis (MG), where serum %Gal(0) is normal, an abnormally low percentage of the anti-TT was agalactosyl. These results imply that in RA and TB a systemic influence is downregulating the galactosylation even of irrelevant IgG. However in SLE and MG antibodies of specificities not studied here must be responsible for the %Gal(0) found in serum. It remains to be seen whether these are the autoantibodies involved in the disease process. PMID- 8722581 TI - Increased utilization of polyreactive B cells during periods of generalized immune activation. AB - This work examines the hypothesis that B cells secreting polyreactive antibodies (antibodies capable of binding to more than one self or foreign antigen) are preferentially utilized during periods of generalized immune stimulation. Four conditions characterized by such stimulation were examined: chronic virus infection, mitogen treatment, autoimmune disease and neonatal repertoire development. In normal adult mice, polyreactive IgM secreting lymphocytes constituted 8-9% of the actively expressed repertoire. Under conditions of generalized immune activation, this frequency increased to 13-19% (p. < .01). Polyreactive IgG secreting B cells, which were present at frequencies of < 0.5% in normal adult mice, were found at freqeuncies of 6-10% in mice with autoimmune disease, chronic virus infection or following mitogen treatment (p. < .001). We postulate that polyreactive lymphocytes are preferentially activated when the immune system is confronted with stimuli inadequately controlled by antigen specific responses. PMID- 8722582 TI - Age-dependent sensitivity to streptozotocin of pancreatic islets isolated from female NOD mice. AB - Streptozotocin (STZ), a selective beta-cell cytotoxin, given in multiple low doses to susceptible mouse strains causes insulin-dependent diabetes mellitus (IDDM) with an autoimmune pathology. Studies in the human suggest that environmental factors such as viruses and certain toxins may modulate the expression of the disease in genetically-prone individuals and the effect may also be age-dependent. Here we have examined the effects of graded, low doses of STZ on beta-cell function and insulin and DNA contents in vitro in cultured islets isolated from female IDDM-prone NOD mice at 4 weeks and at 8 weeks. Results were compared with islets from age and sex-matched non-diabetes prone C57BL/Ks mice. No changes in islet DNA or insulin contents were observed after an acute 30 min exposure to STZ (0, 1.1, 2.2 and 4.4 mM) in the two strains at each of the age groups. However, the DNA content in the NOD mouse islets tended to be lower at 8 weeks, being significant at 1.1 mM STZ. At 4 weeks, islets from NOD mice had a higher insulin content than the control mice but this declined at 8 weeks when it became comparable to the control strain. STZ caused a dose dependent inhibition of islet glucose oxidation rates in all groups. However, at 4 weeks, exposure to 2.2 mM STZ resulted in a significantly greater inhibition in NOD mice than in age-matched control mice. This was reversed at 8 weeks when the islets from NOD mice showed a greater resistance to oxidative impairment than from C57BL/Ks mice. In the presence of 16.7 mM glucose, an inhibitory pattern, similar to the glucose oxidation rate, was also observed for insulin release. In the control mice the relative inhibition of insulin release and glucose oxidation rate was similar at 4 and 8 weeks. These results suggest that islets from the NOD mouse at 4 weeks and prior to insulitis are more sensitive to STZ-induced functional impairment. This enhanced sensitivity suggests that cumulative exposure of diabetes-prone islets to low doses of selective beta cell toxins may be a determinant for later development of IDDM. PMID- 8722583 TI - IgG for intravenous use, autologous serum and plasma induce comparable interleukin-1 receptor antagonist liberation from human mononuclear cells: an in vitro phenomenon depending upon plastic adherence. AB - Immunoglobulin G (IgG) for intravenous use (IVIg) selectively stimulates production of interleukin-1 receptor antagonist protein (IL-1ra) by mononuclear cells in vitro and has been proposed to stimulate IL-1ra production in vivo as part of the therapeutic effect. We tested if IVIg differed from human IgG containing media (i.e., autologous serum (HS) and plasma (HP)) in stimulating IL 1ra release by MNC in vitro and whether IVIg induced a delayed increase in serum levels of IL-1ra. IVIg, 0.01-0.5 mg/ml, increased the IL-1ra liberation from MNC 10-15 times over that of untreated controls. HP and HS (5% v/v) had comparable effects. However, the stimulated IL-1ra liberation was reduced to less than twice the background liberation when fetal calf serum (FCS)-precoated tubes were used. Three days of high-dose IgG infusion had no significant effect on the serum levels of IL-1ra. It is concluded that therapeutic effects of IVIg cannot be ascribed to significant stimulation of IL-1ra production in vivo, as previously suggested, and that the observed stimulation of IL-1ra production in vitro is an epiphenomenon strictly dependent upon adherence of human serum and plasma constituents. PMID- 8722585 TI - Defibrillation shocks over epicardial patches produce sympathetic neural dysfunction in man. AB - INTRODUCTION: The purpose of this study was to determine the effect of direct current (DC) shocks on cardiac sympathetic innervation in humans using I-123 metaiodobenzylguanidine (MIBG) scintigraphy. Decreased efferent sympathetic neural function has been demonstrated following > 10-J DC shocks delivered through epicardial patch electrodes in dogs. To evaluate the effect of DC shocks on cardiac sympathetic innervation in humans, we performed MIBG scintigraphy in 11 patients (ages 46 to 75 years) prior to and after receiving shocks from an implantable cardioverter defibrillator (ICD). METHODS AND RESULTS: This study was performed during an ICD generator change in 7 patients with epicardial patch electrodes and at the time of initial ICD implantation in 4 patients: 2 with epicardial patch electrodes, and 2 with a transvenous ICD system. All patients had spontaneous and inducible ventricular tachycardia. Prior to ICD implantation and remote from any cardioversions or shocks, baseline MIBG and thallium-201 scintigraphy were performed. Repeat MIBG scintigraphy was performed after delivery of ICD shocks and compared with the baseline scans to determine the effect of the shock on sympathetic neural function. The baseline scans revealed focal areas of reduced MIBG uptake in areas of thallium perfusion defects in all patients except the patient without structural heart disease whose scans were normal. Postshock, patients with epicardial patch electrodes who received at least one 24-J shock and had the postshock MIBG scan performed within 4 hours demonstrated no cardiac uptake of MIBG. Two patients with epicardial patch electrodes had no change in the postshock MIBG scans: 1 had a maximal shock of 20 J, and the other had the postshock scan delayed for 11 hours. The 2 patients with a transvenous lead system demonstrated no change in the postshock MIBG scan when compared with baseline. CONCLUSIONS: This study demonstrates that following DC shocks delivered over epicardial patch electrodes, there is diffuse reduction in MIBG uptake that probably represents cardiac sympathetic neural dysfunction that appears to be transient. Sympathetic function does not appear to be affected by shocks delivered over a transvenous lead system. PMID- 8722584 TI - Local electrogram changes in response to a high-voltage intracardiac shock in humans. AB - INTRODUCTION: Transvenous defibrillators may have difficulty sensing ventricular fibrillation following an unsuccessful shock. This study was undertaken to characterize the changes that occur in intracardiac electrograms following a defibrillator shock that may contribute to the failure to redetect arrhythmias. METHODS AND RESULTS: Unipolar and bipolar electrogram recordings were made during sinus rhythm before and following monophasic and biphasic test shocks delivered in random order through a single lead defibrillator system in 15 patients. An additional 14 patients received only a biphasic shock. Electrogram amplitude, dV/dt, and activation recovery time were determined. Following biphasic shocks, unipolar electrogram amplitude decreased by a mean of 19.6% and the bipolar amplitude by a mean of 20.8% (P = NS). The mean dV/dt in the unipolar recordings decreased by 30.7% and in the bipolar recordings decreased by 33.0% (P = NS). Time constants for recovery were approximately 60 seconds. Reduction in dV/dt was greater when the shocking coil was closer to the distal sensing electrode. There was no significant influence of shock waveform (monophasic vs biphasic), recording type (unipolar vs bipolar), or clinical variables on the change in electrogram characteristics. CONCLUSION: Changes in intracardiac electrograms following defibrillator shocks through transvenous leads are seen with shocks delivered in sinus rhythm and may be related to the proximity of the shocking coil to the distal sensing electrode. PMID- 8722586 TI - Developmental effects of d-sotalol on anterograde and retrograde atrioventricular conduction in the rabbit. AB - INTRODUCTION: These experiments investigate the developmental effects of d sotalol on standard electrophysiologic parameters of anterograde and retrograde AV conduction in the rabbit. METHODS AND RESULTS: Using bipolar electrograms and standard pacing techniques, the effects of graded concentrations of d-sotalol on anterograde and retrograde conduction in mature and immature perfused rabbit hearts were compared. Also, a quantitative assessment of the drug's effects on a rate-dependent property of anterograde AV node (AVN) conduction, termed the "recovery process," was compared in mature and immature rabbit hearts. The main developmental electrophysiologic findings of this investigation are: (1) in both the mature and immature rabbit heart, d-sotalol increases the anterograde conduction time and prolongs refractoriness of the AVN, yet the minimal concentrations of d-sotalol that produce these changes are lower in the neonate; (2) d-sotalol increases the anterograde refractory period of the His-Purkinje system in both age groups, but increases anterograde infra-Hisian conduction only in the neonate; (3) 1 x 10-4 M d-sotalol significantly changes the time constant of the AVN recovery process in the neonate, but not in the adult; (4) for retrograde conduction, slow conduction through the AVN (HAmax) and infra-Hisian region (VHmax) are increased by d-sotalol in the neonate, but not in the adult. CONCLUSIONS: The findings of this study illustrate that d-sotalol has different effects on parameters of the developing AV conduction system. This implies that there may be maturational changes in the ionic currents that are responsible for anterograde and retrograde AVN and His-Purkinje conduction. PMID- 8722587 TI - Radiofrequency catheter ablation of the left bundle branch in a canine model. AB - INTRODUCTION: Transcatheter ablation of the left bundle branch may be considered for management of selected macroreentrant ventricular tachycardias. Left bundle ablation can also change the sequence of left ventricular contraction and may simulate pacing in hypertrophic obstructive cardiomyopathy. The purpose of this study was to determine electrophysiologic and anatomic parameters for successful selective transcatheter left bundle ablation in a canine model. METHODS AND RESULTS: A catheter was advanced to the left ventricular apex and the tip deflected toward the septum, until a discrete left bundle potential (LBP) was found. Radiofrequency (RF) energy was then applied until left bundle branch block or complete AV block occurred. In 29 (85%) dogs, an LBP (mean LBP-V 16 +/- 3 msec; range 10 to 20 msec) was identified resulting in successful left bundle ablation. In 5 (15%) dogs, a similar potential (mean potential-V 28 +/- 4 msec; P = 0.001 vs LBP-V) was identified, but RF energy application produced complete AV block. The A:V electrogram ratio at the successful LBP ablation site was < 1:10 in all 29 dogs successfully ablated, but only 2 (40%) of 5 dogs in the unsuccessful group (P = 0.0017). In 4 successfully ablated dogs, the right bundle potential was mapped and complete AV block was created by RF energy application, confirming that the left bundle was completely ablated. In 9 dogs, the left bundle and AV junction were sequentially ablated with 1 lesion at each site. Postmortem examination showed 2 discrete lesions 1.2 +/- 0.7 cm apart. CONCLUSIONS: Selective transcatheter left bundle ablation was successfully guided by the LBP. The distance between the AV junction and the main left bundle was 1.2 cm in this canine model. An A:V ratio < 1:10 and an LBP-V time < 20 msec appear to minimize the risk of AV block. Prudent use of similar techniques may cure macroreentrant ventricular tachycardias and reduce the need for permanent pacing in hypertrophic obstructive cardiomyopathy. PMID- 8722588 TI - Anisotropy, fiber curvature, and bath loading effects on activation in thin and thick cardiac tissue preparations: simulations in a three-dimensional bidomain model. AB - INTRODUCTION: A modeling study is presented to explore the effects of tissue conductivity, fiber orientation, and presence of an adjoining extracellular volume conductor on electrical conduction in cardiac muscle. Simulated results are compared with those of classical in vitro experiments on superfused thin layer preparations and on whole hearts. METHODS AND RESULTS: The tissue is modeled as a three-dimensional bidomain block adjoining an isotropic bath. In the thin layer model, the fibers are assumed parallel. In the thick block model, fiber rotation, curvature, and tipping are incorporated. Results from the thin layer model explain experimental observations that the rate of rise of the entire action potential upstroke is faster and the magnitude of the extracellular potential is smaller across fibers than along fibers in a uniformly propagating front. The simulation identified that this behavior only arises in tissue with unequal anisotropy in the two spaces and adjoining an extracellular bath. Simulated conduction and potential distributions in the thick block model are shown to well approximate experimental maps. The potentials are sensitive to changes in the fiber orientations. A slight 5 degrees tipping of intramural fibers out of the planes parallel to the epicardium and endocardium will lead to an asymmetry of the magnitudes of the positive regions. In addition, the introduction of fiber curvature leads to more realistic isochrone and extracellular potential distributions. The orientation of the central negative region of the extracellular potential is shown to be determined by the average of the fiber direction at the plane of pacing and the plane of recording. CONCLUSIONS: The simulations demonstrate the sensitivity of spread of activation and potential time courses and distributions to the underlying electrical properties in both thick and thin slabs. The bidomain model is shown to be a useful representation of cardiac tissue for interpreting experimental data of activation. PMID- 8722589 TI - Bezold-Jarisch-like phenomenon induced by radiofrequency ablation of a left posteroseptal accessory pathway via the coronary sinus. AB - We report a case of asystole induced by radiofrequency (RF) ablation via the coronary sinus in a 35-year-old man suffering from symptomatic left posteroseptal accessory pathway. RF application provoked progressive slowing of the sinus rhythm, disappearance of the preexcitation, and an 8-second period of asystole followed by atrial fibrillation. The causal mechanism proposed is a strong stimulation of vagal afferent pathways linked with sensory endings of the inferoposterior myocardial wall leading to a Bezold-Jarisch-like phenomenon. PMID- 8722590 TI - Autonomic dysfunction after catheter ablation. AB - Autonomic dysfunction may occur as a consequence of radiofrequency (RF) catheter ablation of a variety of supraventricular tachycardias. Effects suggestive of autonomic dysfunction that may be seen acutely during the ablation procedure include sudden profound slowing of the sinus rate or transient AV block. These abnormalities may occur during application of RF current, typically along the tricuspid or mitral annulus, at sites distant from both the sinus and AV nodes; they resolve quickly when RF current delivery is terminated. The most common long term indication of autonomic dysfunction after ablation is inappropriate sinus tachycardia. This complication, rarely a lasting significant clinical problem, is seen after AV node modification and after ablation of accessory pathways. It usually resolves within several months. The mechanism appears to be loss of parasympathetic influence on the sinus node. Autonomic dysfunction after ablation of ventricular tachycardia has not yet been described, but could occur as newer catheter technologies capable of producing larger lesions are perfected. PMID- 8722591 TI - To fumble flutter or tackle "tach"? Toward updated classifiers for atrial tachyarrhythmias. AB - Aristotle proposed in his short work, The Categories, that a definition is a statement of a thing's essential nature, and the essence of a thing are those of its properties that cannot change without losing its identity. But Aristotle was not faced with the flux of new information that confronts modern medicine. Nowadays, the argot of a discipline arises organically at the intersection of a given state of empiric knowledge and the exigencies of present scientific discourse. Thus, when the only treatment for a regular, narrow QRS complex tachycardia was digitalis glycosides or vasopressor infusion, the term "PAT" ("paroxysmal atrial tachycardia") seemed adequate, at least to distinguish it from ventricular tachycardia. We now prefer the term "PSVT" (paroxysmal supraventricular tachycardia) because we understand that most such tachycardias are not in truth "atrial" but involve the AV node and/or an accessory AV connection, and because we wish to report on the results of treatment specific to each of the subcategories of "PSVT." Similarly, as our knowledge of atrial arrhythmias has grown and especially as we need to describe the outcome of new interventional approaches to therapy, it may be prudent to use a nomenclature for atrial tachyarrhythmias that is based on the geometry of the tachycardia substrate, the relationship of that substrate to atrial anatomy, and the type of atrial lesions required to abolish that substrate. PMID- 8722592 TI - What is the mechanism of the change in QRS morphology during tachycardia? PMID- 8722593 TI - Surgical therapy of ventricular tachyarrhythmias in patients with coronary artery disease. PMID- 8722602 TI - Modulation of in vitro cytokine release from human leukemic mast cells (HMC-1) by glucocorticoids. AB - Mast cells are well known effector cells not only in allergic but also in diverse acute and chronic inflammatory diseases. We have shown previously that these cells produce a broad spectrum of cytokines which might contribute to mast cell dependent pathology. In the present study, we have investigated the influence of four potent glucocorticoids, methylprednisolone-aceponate, methylprednisolone-17 propionate, prednicarbate, and betametasone valerate (10(-5) M-10(-9) M), on the IL-1 beta, IL-3, IL-8, and tumor necrosis factor alpha secretion of the HMC-1 mast cell line as measured by ELISA. All four glucocorticoids caused a comparable dose- and time-dependent inhibition of cytokine release from HMC-1 cells stimulated for 24 h with phorbol 12-myristate 13-acetate 25 ng/ml and calcium ionophore 2 x 10(-7) M. These results shed further light on the mechanisms involved in antiinflammatory effects of glucocorticoids in allergic inflammation. PMID- 8722603 TI - Effect of fumaric acid, its dimethylester, and topical antipsoriatic drugs on epidermal differentiation in the mouse tail model. AB - Fumaric acid, fumaric acid dimethylester, and the dithranol derivative C4-lactone were studied in the mouse tail test to evaluate their effects on epidermal cell differentiation compared with other topical antipsoriatic drugs, such as betamethasone, calcipotriol, and dithranol. Mouse tails were treated for 2 weeks and longitudinal histological sections prepared of the tail skin. The length of the orthokeratotic regions (stratum granulosum) was measured on 10 sequential scales per tail and expressed as percentage of the full length of the scale. In addition, epidermal thickness was measured and the efficacy of the various compounds evaluated. In comparison to 2% salicylic acid ointment, all tested compounds except fumaric acid significantly (p < or = 0.05) increased the proportion of the orthokeratotic region. C4-lactone and calcipotriol were less effective than dithranol, fumaric acid dimethylester only moderately influenced cell differentiation, and betamethasone showed the least potent effect. Dithranol was the most potent substance inducing orthokeratosis without increasing epidermal thickness. PMID- 8722604 TI - Evaluation of the effect of zinc acetate on the stratum corneum penetration kinetics of erythromycin in healthy male volunteers. AB - Erythromycin with or without additional zinc acetate is used topically in the treatment of acne vulgaris. A potential effect of zinc on the stratum corneum penetration of erythromycin was investigated in human volunteers. Skin surface washings and tape strippings from the skin of the back were collected after drug applications in 12 subjects for quantification of erythromycin levels. Zinc acetate increased the amount remaining on the back skin at 6 h after application from 40 +/- 19 to 56 +/- 15% of the dose and, vice versa, reduced the amount in stratum corneum strips from 22 +/- 7 to 18 +/- 7%, both with statistical significance. The effect varied with body region. Zinc acetate thus provided to prolong the residence time of erythromycin on the skin. PMID- 8722605 TI - Cutaneous permeability barrier repair following various types of insults: kinetics and effects of occlusion. AB - Previous studies have shown that acute disruption of the cutaneous permeability barrier by acetone results in an initial rapid phase of repair followed by a later, slower phase. In the present study, we demonstrate that manipulations which disrupt the barrier by other mechanisms, such as tape stripping or detergent treatment, have a similar pattern of barrier repair. In all three models, the return of lipid to the stratum corneum parallels the normalization of barrier function, and occlusion immediately after disrupting the barrier blocks both the return of lipid and the normalization of function. Moreover, occlusion beginning 6-8 h following barrier disruption blocks the late, slower phase of repair, indicating that the late phase can be inhibited independently of the initial phase. Lastly, both severe and relatively minor perturbations of the barrier elicit a repair response with a similar kinetic pattern. In summary, the present study demonstrates that barrier repair responses are similar regardless of the etiology or extent of barrier disruption. PMID- 8722606 TI - 0.05% clobetasol 17-propionate cream and ointment but not the corresponding 0.1% triamcinolone acetonide preparations increase skin surface roughness: a possible dissociation of unwanted epidermal and dermal effects. AB - Repeated open application of clobetasol 17-propionate cream and ointment to normal skin over a period of 6 weeks induced an increase in skin surface roughness as assessed by profilometry (p < 0.05), while 6 weeks' application of triamcinolone acetonide cream and ointment did not. The increase in skin roughness with clobetasol 17-propionate cream turned out to be greater than with ointment containing identical amounts of clobetasol 17-propionate (p < 0.05). A clear-cut correlation between increase of skin surface roughness and skin thickness as assessed by high-frequency ultrasound could be demonstrated only with clobetasol 17-propionate cream and ointment. PMID- 8722607 TI - Human skin irritation studies of a lecithin microemulsion gel and of lecithin liposomes. AB - Soybean lecithin microemulsion gels offer promising features for the possible use as matrices in transdermal therapeutic systems. In order to assess the skin irritancy potential of the gel, acute and cumulative irriation tests were performed in human subjects in vivo using as comparison an unilamellar soybean lecithin liposome preparation and the solvent isopropyl palmitate (IPP). Acute irritation was tested in 151 volunteers in a 48-hour patch test, whereas cumulative irritation was assessed in a 21-day human repeated insult patch test in 20 volunteers. In the acute irritation test, discrete irritation (erythema only) developed with the gel in 2 subjects (1.3%), with the liposomes in 3 subjects (2.0%), and with IPP in 2 subjects (1.3%). For the assessment of cumulative irritation, the IT50 (irritation time of 50% of the test population) was calculated. IT50 was 13 days for the gel, 14 days for the liposomes and 17 days for IPP. This study shows a very low acute and a low cumulative irritancy potential for the soybean lecithin microemulsion gel making it a candidate matrix for transdermal therapeutic systems also under toxicological aspects. PMID- 8722608 TI - In vitro/in vivo difference in enhanced skin permeation of nicardipine hydrochloride by the 1-menthol-ethanol system. AB - The in vitro/in vivo difference in enhanced skin permeation of nicardipine hydrochloride (NC) by simultaneous use of 1-menthol and ethanol (MEW system, 1 menthol:ethanol:water = 5:40:55) was investigated in hairless rats. First, the cutaneous blood flow clearance (clearance from skin to blood flow) of NC per unit area of skin (CLCB/A), which was comparable to the permeability coefficient across skin (PC), was calculated from intracutaneous and intravenous injection data using the deconvolution method; the value was 1.67 microliters/h/cm2. Two formulations containing NC and the MEW system, solution (SOL) and 15% hydroxypropyl cellulose gel (GEL), were used for in vitro and in vivo permeation experiments. The in vitro PC of NC via excised skin from SOL (23.3 microliters/h/cm2) was significantly higher than CLCB/A, and that from GEL (1.48 microliters/h/cm2) was similar to CLCB/A. Consequently, the steady-state concentration of NC in skin during in vivo application of SOL was 6.6 times higher than corresponding in vitro data. In vivo PCs from SOL and GEL were however significantly lower than CLCB/A. These results may be explained by the findings, using a laser Doppler flowmeter, that cutaneous blood flow was decreased by the application of MEW. PMID- 8722609 TI - Cyclosporine A inhibits tPA mRNA transcription in A431 cell line. AB - Cyclosporine A (CyA), a well established treatment of psoriasis, is a highly lipophilic cyclic undecapeptide mainly used for its immunosuppressive properties and exerting a wide spectrum of biological activities including fungicidal antiproliferative and anti-inflammatory effects. Plasminogen activators (PA), urokinase (UK, M(r) 55,000) and tissue type plasminogen activators (tPA, M(r) 74,000), physiologically catalyze the conversion of the plasminogen to the wide spectrum proteinase plasmin. UK and tPA are involved in cell growth, differentiation and migration. It has recently been shown that psoriatic epidermis is provided with abnormal tPA-dependent PA activity and that in lesional epidermis elevated tPA mRNA levels are present. It has been suggested that the tPA-dependent PA activity is a marker of disease activity and is reversible with different topical and systemic treatments. In this preliminary study we investigate the effect of CyA on the tPA mRNA transcription on A431 keratinocytes cell line. Subconfluent A431 cell cultures have been treated with CyA at in vivo relevant concentrations (10, 7.5, 5 micrograms/ml) for 48 h. Northern blot analysis of total RNA extracted from cultured A431 cell line has been performed for detecting tPA mRNA. mRNA for tPA has been detected in the control samples whereas an evident decrease of tPA mRNA expression has been detected in the CyA-treated samples. These data suggest that CyA could have an effect in clearing psoriatic lesions also modulating the abnormal plasminogen activation i.e. tPA-dependent serinoproteinase activity. PMID- 8722610 TI - Communicable disease report: Salmonella indiana. PMID- 8722611 TI - AIDS and HIV-1 infection in the United Kingdom: monthly report. PMID- 8722612 TI - Mucin (Muc-1) expression is differentially regulated in uterine luminal and glandular epithelia of the baboon (Papio anubis). AB - Changes in the organization and composition of apical cell surface glycoconjugates accompany the transition of luminal epithelial cells from the prereceptive state of the uterus in many species. In spite of the biological and clinical significance of this process, few molecular markers have arisen as useful predictors of uterine receptivity. Recent studies in mice demonstrate that the transmembrane mucin glycoprotein, Muc-1, is abundantly expressed at the apical surface of luminal epithelia under most conditions and is invariably reduced in receptive uteri. These and other observations have led to the suggestion that mucins serve an antiadhesive role and function to maintain a nonreceptive uterine state. A pan-species Muc-1 specific antibody recognizing a peptide motif conserved in the cytoplasmic domain of Muc-1 was used to examine the temporal and spatial expression of cell-associated Muc-1 in baboon uteri under a variety of conditions, including the pre- and perimplantation periods. Muc-1 expression was not driven by estrogen influences alone, but required progesterone action. In animals exposed to both steroids, Muc-1 was expressed at low moderate levels in epithelia of the basalis and functionalis regions. The highest expression of Muc-1 was detected in surface epithelium of the preimplantation phase, i.e., up to Day 8 (Day 0 = day of ovulation), or in ovariectomized animals receiving a steroid hormone regime that mimicked this phase (14 days of estrogen priming followed by 7 days of estrogen plus progesterone). Continued exposure to both hormones, i.e., as seen at Days 10-12 or in ovariectomized baboons given 14 days of estrogen plus progesterone treatment after estrogen priming, resulted in marked reduction of Muc-1 expression in the surface epithelium; however, staining patterns in the glandular epithelium were unchanged by this treatment. The expression of Muc-1 on the surface epithelium during the prereceptive phase was associated with the presence of both estrogen and progestin receptors in these epithelia. Muc-1 expression was reduced by neither antiestrogen treatment during the prereceptive stage nor antiprogestin treatment through to the receptive phase. Furthermore, persistent Muc-1 expression in the functionalis and basalis epithelium correlated with expression of progestin receptors. Thus, Muc-1 expression appeared to be progesterone-dependent rather than estrogen-dependent. It is concluded that Muc-1 expression in surface epithelium serves as a marker of the prereceptive phase in the baboon and that loss of Muc-1 from surface epithelium correlates with generation of a receptive uterine state. PMID- 8722613 TI - Effect of developmental age or time after transplantation on Sertoli cell number and testicular size in inbred Fischer rats. AB - The objectives were to establish the developmental age of Fischer rats at which the Sertoli cell number is stabilized, to establish the normal reference plateau number of Sertoli cells for evaluation of testes after transplantation, and to determine whether the developmental pattern establishing Sertoli cell proliferation and stability are similar between intact and transplanted testes. Sertoli cell number was determined at ages 1--120 days in intact rats and at various times (10-90 days) after transplantation of prenatal or neonatal tests. Tests were fixed by vascular perfusion or by immersion with 2% glutaraldehyde and and immersion in 1% osmium and were embedded in Epon 812. Sections and serial sections were cut at 0.5 micrometer to determine the Sertoli cell nuclei volume density and the volume of an individual Sertoli cell nucleus by brightfield microscopy or at 20 micrometers to determine the maximum height and width of nuclei. A correction factor was calculated for intact (0.663 +/- 0.025) or for transplanted (0.558 +/- 0.029) testes to determine the volume of a single Sertoli cell nucleus from height and width measurements. In intact testes, Sertoli cell numbers significantly increased to Day 20 but were not different between 15 and 90 days. Sertoli cell number in prenatal or neonatal transplanted testes increased to 20 or 30 days posttransplantation and then stabilized to Day 60 or 90. There was no difference in the plateau number of Sertoli cells per rat between prenatal and neonatal testes. Sertoli cells in 10-day- and 30-day transplanted testes incorporated 3H-thymidine when placed in culture. A few tubules had complete spermatogenesis at 90 days posttransplantation, indicating that Sertoli cells in some of these tubules were functional. Leydig cell structure appeared to be normal. Leukocytic infiltration of testes was not observed in intact rats or in rats receiving neonatal testes. Although transplanted testes showed a delay in reaching the plateau value for Sertoli cell number per testis and although the value reached was lower, the developmental pattern of Sertoli cell proliferation in transplanted testes was similar to that in intact rats. PMID- 8722614 TI - Effect of hypophysectomy, sex of host, and/or number of transplanted testes on Sertoli cell number and testicular size of syngeneic testicular grafts in Fischer rats. AB - One or more neonatal testicular grafts were transplanted for 60-65 days into young adult inbred Fischer rats to determine the effect of hypophysectomy, sex of host, and/or the number of transplanted testes on testicular size and Sertoli cell number. All host rats had been castrated or ovariectomized and some were hypophysectomized as well. At the end of the treatment, testes were fixed and embedded in Epon before histologic sections (0.5 micrometer or 20 micrometers) were evaluated by stereology. Testicular grafts placed in castrated adult male rats with intact pituitaries weighed more (p < 0.01) and had more (p < 0.01) Sertoli cells than those placed in hypophysectomized hosts. Testicular grafts that were recovered from hypophysectomized rats 34 days posttransplantation and placed in pituitary-intact males for 30 day had larger (p < 0.05) parenchymal weights and more Sertoli cells than did testes re-transplanted into hypophysectomized rats. However, this delayed period of Sertoli cell proliferation id not extend to 65 days of hypophysectomy. When two or four testes were transplanted into castrated males or ovariectomized female hosts for 65 days, there was no difference in the graft weights or Sertoli cell numbers between sexes. Four transplanted testes per rat produced more (p < 0.01) total testicular parenchyma and a greater (p < 0.01) number of Sertoli cells per testis than did two tests regardless of sex of the host. This model has shown that the period of Sertoli cell proliferation can be delayed by hypophysectomy, that Sertoli cell number can be influenced by endogenous hormones, and that a major component in regulation of testicular size is at the level of the testis in this model. Hence, this model should facilitate study of experimental endocrine manipulation control and potential experimental intervention to increase Sertoli cell number and testicular size. PMID- 8722615 TI - Characterization of ovine stem cell factor messenger ribonucleic acid and protein in the corpus luteum throughout the luteal phase. AB - Stem cell factor (SCF) is a growth factor known to have profound effects on the proliferation, migration, differentiation, and survival of numerous cell types, including those of the ovary. The objectives of the present study were to identify and characterize expression of this growth factor in the ovine corpus luteum (CL). A 952-bp cDNA was amplified from Day 3 (Day 0 = estrus) ovine luteal total cellular (tc) RNA by reverse transcriptase-polymerase chain reaction and determined to encode SCF. Northern analysis of Day 10 luteal poly(A)+ RNA indicated one major transcript of approximately 6.5 kb. SCF mRNA was localized within Day 3 and Day 10 CL by in situ hybridization and was expressed throughout luteal tissue on both days examined. To asses expression throughout the luteal phase, SCF mRNA was quantified by ribonuclease protection assay in tcRNA collected on Day 3, 7, 10, 13, and 16; values did not differ across days (p > 0.10). Similarly, SCF mRNA was quantified in tcRNA isolated from pools of Day 10 large and small steroidogenic cells (n = 4 and 3, respectively); levels did not differ (p > 0.10) between cell types. In addition, SCF protein was detected in CL on Days 3 and 10, and was expressed in a cell-specific manner in cells with morphological characteristics of large and small luteal cells. These data indicate that SCF may be involved in communication among steroidogenic cells and/or between steroidogenic and nonsteroidogenic cells of the CL. PMID- 8722616 TI - Interphase chromosome arrangement in Sertoli cells of adult mice. AB - Sertoli cells of adult male laboratory mice were examined with a number of banding techniques and by nonradioactive in situ hybridization applying different repetitive DNA probes. All banding methods revealed the typical features of mouse Sertoli cells, i.e., a central nucleolus, usually with two chromocenters associated at diametrically opposed sides in which the centromeric regions of the chromosomes are clustered. Silver staining as well as in situ hydridization with rDNA labeled part of the chromocenters and the nucleolus, indicating transcriptional activity of at least some of the nucleolus organizer regions. In situ hybridization with X- and Y-specific DNA probes showed both sex chromosomes to be undercondensed in Sertoli cells This decondensation suggests expression of sex chromosomal genes in Sertoli cells. While the X chromosome appeared to occupy a central position near one of the chromocenters, the Y chromosome was found at the periphery of the nucleus in the majority of cells. Hybridization with telomeric sequences resulted in strong labeling of the chromocenters and dispersed signals at the nuclear periphery. PMID- 8722617 TI - Sexual differences in the diurnal changes of tuberoinfundibular dopaminergic neuron activity in the rat: role of cholinergic control. AB - A diurnal change of the tuberoinfundibular dopaminergic (TIDA) neuron activity in ovariectomized rats treated with or without estrogen was recently reported, and the endogenous cholinergic system may be responsible for its induction. Whether a similar rhythm exists in intact female or male rats was the focus of this study. TIDA neuron activity was assessed by measurement of the precursor or metabolite of dopamine in the median eminence by HPLC with electrochemical detection. In intact female Sprague-Dawley rats, diurnal changes in TIDA neuron activity were observed during all stages of the estrous cycle, i.e., proestrus, estrus, and diestrus 1, and they were nearly identical. No such rhythm, however, was observed in intact male rats. Castration alone decreased and increased basal TIDA neuron activity in female and male rats, respectively, and estrogen treatment increased the activity in both sexes. The diurnal changes in TIDA neuron activity, however, were observed only in the female rats, not the male rats, regardless of castration or estrogen treatment. Serum prolactin levels in the male rats exhibited no diurnal changes either, irrespective of whether the animals had been castrated or treated with estrogen. Central administration of mecamylamine (1 micrograms/3 microliters per rat, i.c.v.), a nicotinic receptor antagonist, in the morning (at 1000 h) or afternoon (at 1500 and 1700 h) induced a differential effect on the TIDA neuron activity of intact female rats, i.e., no effect in the morning and stimulation in the afternoon; it had no effect on the TIDA neuron activity in the male rats regardless of the injection time. In castrated male rats treated with estrogen, however, mecamylamine treatment further increased TIDA neuron activity, but still with no diurnal difference. In summary, a sexual difference in the diurnal changes of TIDA neuron activity was observed, and these changes may be under differential control by the cholinergic system. PMID- 8722618 TI - Human sperm plasma membrane progesterone receptor(s) and the acrosome reaction. AB - Progesterone initiation of te human sperm acrosome reaction (AR) includes stimulation of a transient Ca2+ influx and a transient Cl- efflux. A role for one or more plasma membrane receptors has been suggested, but, except for evidence supporting the involvement of a sperm GABAA-like receptor/Cl- channel, there is little information about possible receptor identity. Here, we attempt to identify plasma membrane progesterone receptors in human sperm using a mouse monoclonal antibody (mAb-C262) raised against the C-terminal steroid-binding domain of the human intracellular progesterone receptor. C-262 inhibited the progesterone initiated AR in a dose-dependent manner. Maximum inhibition was 77% as detected by fluorescein isothiocyanate (FITC)-concanavalin A (conA). Motility was unaffected. A control mouse mAb (h-151) raised against the human estrogen receptor did not inhibit the progesterone-initiated AR. Western blotting with C 262, but not with h-151, detected a major sperm protein band of 50-52 kDa. In indirect immunofluorescence localization studies, live and ethanol-fixed uncapacitated sperm and fixed capacitated sperm incubated with C-262, but not with h-151, displayed fluorescence at the equatorial segment region of the sperm head plasma membrane. In spectrofluorometric studies using capacitated sperm loaded with the Ca2+ probe Fura-2 or the Cl- probe MEQ, C-262 but not h-151 inhibited both Ca2+ influx and Cl- efflux. These ion fluxes could be due to the binding of progesterone to two different receptor/channels or to its binding to one and cross talk with the other. Our results strongly support the involvement of sperm plasma membrane receptors in the progesterone-initiated AR and provide a candidate for one such receptor. PMID- 8722619 TI - Influence of hypothalamus and ovary on pituitary function in transgenic mice expressing the bovine growth hormone gene and in growth hormone-deficient Ames dwarf mice. AB - Female transgenic mice expressing the bovine (b) growth hormone (GH) gene exhibit reduced fertility, and GH-deficient dwarf mice are sterile. However, in these animals, the influence of the hypothalamus and ovary on pituitary functions is poorly understood. To address this issue, female Ames dwarf mice received twice daily injections of either vehicle or bGH (25 micrograms/injection per mouse) for 12 days. Vehicle-injected normal female siblings served as controls. On Day 7, these mice and adult female transgenic mice expressing the bGH gene with mouse metallothionein-I promoter, as well as their normal littermates, were ovariectomized. Two experiments were conducted with each group of mice: on Day 8 after ovariectomy of transgenic mice and their normal siblings, and on Day 4 after ovariectomy of dwarf mice and their normal siblings. In experiment I, mice were primed with 0.5 micrograms estradiol benzoate (EB) in oil and 24 h later received injections of 5 micrograms EB/100 g BW. The next day, these mice received injections of either saline or GnRH (1 ng/g BW), and 15 min later blood was obtained via heart puncture under anesthesia. In experiment II, ovariectomized mice either received injections of oil or were primed with 0.5 micrograms EB and 24 h later treated with 10 micrograms EB/100 g BW in oil. On the next day, blood was obtained as in experiment I. In transgenic mice and in their normal littermates, plasma bGH, FSH, LH, and prolactin levels were measured. In dwarf mice and in their normal siblings, plasma LH and insulin-like growth factor (IGF)-I levels were determined. Bovine GH was secreted in transgenic mice, but this hormone was not detectable in normal siblings. Expression of the GH gene resulted in attenuated (p < 0.001) FSH response to GnRH treatment, while LH response was similar to that in controls. The ovariectomy induced increases in plasma FSH (p < 0.001) and LH (p < 0.05) levels were decreased in transgenic mice. The absolute circulating FSH levels were also decreased (p < 0.001) in estrogen-treated transgenic mice, while plasma LH level were similar in normal mice and in mice bearing the bGH gene. The ovariectomized transgenic mice were hyperprolactinemic. In ovariectomized dwarf mice, IGF-I was not detectable, and GH treatment induced its secretion. In these mice, plasma LH concentrations were reduced (p < 0.05), and GH treatment increased (p < 0.01) LH secretion. In oil-injected ovariectomized dwarf mice, the plasma LH response to GnRH treatment was reduced (p < 0.001), and GH administration normalized this response. The negative feedback effect of estrogen on LH secretion was decreased (p < 0.025) in dwarf mice. These indicate that the alterations in gonadotropin secretion are dependent on the duration of exposure of neuroendocrine system to bGH and that bGH alters the effects of GnRH and estrogen on gonadotropin secretion in these two widely divergent experimental mouse models. PMID- 8722620 TI - Surface localization of P34H an epididymal protein, during maturation, capacitation, and acrosome reaction of human spermatozoa. AB - During epididymal transit, spermatozoa acquire new surface antigens that are involved in the acquisition of their fertilizing ability. We have previously described a 34-kDa (P34H) human epididymal sperm protein that shows antigenic and functional homologies with the hamster P26h. P34H is localized on the acrosomal cap of human spermatozoa and has been proposed to be involved in the interaction with the zona pellucida. The aim of this study was to document the expression of P34H on the sperm surface during transit along the male and female genital tracts. Immunohistochemical techniques were performed on human testes and epididymides by means of an antiserum specific for P34H. No labelling was detected on those spermatozoa found within the seminiferous tubules or in the vasa efferentia. P34H first appeared in the caput epididymidis and was restricted to the acrosomal cap. Signal intensity then increased considerably from the proximal corpus to the cauda region of the epididymis. After ejaculation, the same pattern of P34H distribution was observed, but the intensity was much lower than that characterizing the cauda epididymal spermatozoa. Strong labeling was restored after incubation in B2 medium and was maximal after 5 h of capacitation. After acrosomal exocytosis induced by a Ca2+ ionophore, the percentage of P34H labeled spermatozoa decreased proportionally to the number of acrosome-reacted spermatozoa as determined by Pisum sativum-fluorescein isothiocyanate (FITC) labeling. P34H appeared to be strongly anchored to the sperm plasma membrane during epididymal transit as indicated by the requirement for detergent to extract this surface antigen from ejaculated spermatozoa. This confirms the importance of P34H binding to the sperm plasma membrane during epididymal maturation. We have previously proposed that P34H is involved in sperm-zone pellucida interaction. The appearance and accumulation of P34H on the sperm plasma membrane during epididymal maturation, followed by its inaccessibility associated with ejaculation, its unmasking during capacitation, and finally its elimination after the acrosome reaction, are in agreement with te proposed function of this sperm antigen. PMID- 8722621 TI - Cases of human infertility are associated with the absence of P34H an epididymal sperm antigen. AB - The interaction of spermatozoa with the zona pellucida is a critical step of fertilization. Specific sperm surface proteins involved in this process can be added or modified during epididymal transit. We have previously described a 34 kDa human epididymal sperm protein (P34H) that we proposed to be involved in sperm-zona pellucida interaction. In this study, Western blot analysis were performed to determine the level of P34H protein present on the spermatozoa of 16 men with idiopathic infertility. These levels were compared with the amount of P34H protein found in men of proven fertility. In addition, a sperm-zona pellucida binding assay was performed with spermatozoa from fertile and infertile men. Spermatozoa obtained from different semen samples from a given individual had similar P34H levels. However, the amount of P34H varied from one man to another. Nine of 16 infertile men had a P34H level that was less than 30% of the normal value based on a population of fertile men, while the remaining 7 males were in the normal range. Sperm from infertile subjects with a normal P43H determination bound to zonae pellucidae as efficiently as those from controls. However, spermatozoa from subjects with a low amount of P34H exhibited a dramatic diminution in their ability to interact with zonae pellucidae. Our results show that the quantity of the epididymal protein P34H varied from one male to another and that low levels of this epididymal sperm protein are associated with certain cases of idiopathic infertility. Results are discussed with regard to the function of human epididymis in sperm maturation. PMID- 8722622 TI - Progestin-induced puberty and secretion of luteinizing hormone in heifers. AB - Two experiments were designed to investigate the endocrine mechanisms by which progestin administration induces puberty in heifers. In experiment 1, prepubertal heifers were randomly assigned by age to receive either a single norgestomet implant for 10 days (NORG; n = 15) or to serve as unimplanted controls (CONT; n = 14). Serial blood samples were obtained on Days -0.5, 8.5, and 10.5 (Day 1 = day of implant insertion) and were analyzed for concentrations of LH. On days 9 and 11, 4 heifers in each treatment were slaughtered, and the reproductive tracts were obtained. Weekly progesterone analyses were performed to estimate the day of puberty in heifers not slaughtered. Puberty was induced in 6 of 7 heifers in the NORG treatment, resulting in an earlier (p < 0.05) day of puberty in the NORG than in the CONT treatment. The frequency of LH pulses was higher (p < 0.05) on Days 8.5 and 10.5 in the NORG as compared to the CONT treatment. Although no difference (p > 0.10) was observed between treatments in follicular development, uterine weight was greater (p < 0.05) in NORG than in CONT heifers on Day 11. In experiment 2, prepubertal heifers (n = 47) were administered either 1 (1NORG; n = 16), 3(3NORG; n = 16), or 0 (CONT; n = 15) norgestomet implants for 10 days, and serial blood samples were obtained as described for experiment 1. Transrectal ultrasonography was used to determine the diameter of the largest follicle on Day 9. Plasma samples were obtained after each serial sample collection period and were analyzed for estradiol concentrations. Puberty was induced in 75% (12 of 16) and 81% (13 of 16) of heifers in the 1NORG treatments, respectively. Four heifers in the 1NORG treatment, from which serial blood samples were collected, ovulated before removal of the progestin implant, and the LH data for this treatment were deleted. In the 3NORG treatment, LH pulse frequency was suppressed (p < 0.05) on Day 8.5, but was greater (p < 0.05) 12 h after removal of the progestin (Day 10.5) than in the CONT treatment. We conclude that progestin administration hastens puberty by increasing LH secretion after progestin withdrawal and propose that progestin administration induces puberty by accelerating the peripubertal decrease of estradiol negative feedback on LH secretion. PMID- 8722623 TI - Temporal surge of glycosyltransferase activities in the genital tract of the hamster during the estrous cycle. AB - Mammalian spermatozoa must undergo maturational changes between the events of mating and fertilization. These biochemical and functional alterations, collectively termed capacitation, take place as spermatozoa traverse the female reproductive tract. The preparatory biochemical changes include removal, modification, and reorganization of sperm surface molecules. Although details of all the changes are not known, lectin binding studies have provided evidence suggesting that carbohydrate moieties of sperm surface glycoproteins are modified during capacitation. In an attempt to gain insight into the potential modifications of sperm plasma membrane glycoproteins, we quantified glycoprotein modifying enzyme activities in the uterine and oviductal fluid of the hamster during the 4 days of the estrous cycle. These enzymes are known to modify existing glycoproteins, either by adding sugar residues (glycosyltransferases) or by removing terminal sugar residues (glycosidases). Data from these studies showed that 1) levels of all glycosyltransferase activities assayed (sialyltransferase, fucosyltransferase, galactosyltransferase, and N acetylglucosaminyltransferase) were negligible in the uterine fluid at the onset of ovulation (Day 1) but sharply increased preceding ovulation (Day 4); 2) levels of the four glycosyltransferase activities assayed were higher in the oviductal fluid at the onset of ovulation (Day 1) and then gradually decreased through the remainder of the estrous cycle (Day 2 to Day 4); 3) levels of all glycohydrolase activities (acidic alpha-D-mannosidase, beta-D-galactosidase, beta-D glucuronidase, beta-D-glucosaminidase, and alpha-L-fucosidase) and protein in the uterine and oviductal fluids did not vary widely during the 4 days of the cycle. These results demonstrate a temporal surge of glycosyltransferase activities in the genital tract fluids of the hamster. The temporal changes in the glycoprotein modifying enzymes may have an effect on the glycosylation of sperm plasma membrane and zona pellucida glycoproteins at the site of fertilization or may alter the surface glycoproteins of the fertilized egg in the uterus prior to implantation. PMID- 8722624 TI - Chemical cues are necessary but insufficient for reproductive activation of female pine vole (Microtus pinetorum). AB - Among various arvicoline rodents, reproduction is influenced to varying degrees by social factors, including behavioral or chemical cues. Since previous research suggested that chemosignals from adult males reproductively activate female pine voles (Microtus pine-torum), we sought to determine specifically what types of stimuli promote the activation response. In these experiments, female were exposed to unfamiliar adult males, or to some combination of cues from males, or were housed alone. Using uterine mass as a measure of reproductive activation, we found that females were not activated by exposure either to male urine by itself or to male-soiled bedding by itself, but full contact with a male clearly resulted in heavier uteri. Females whose vomeronasal organs were surgically excised failed to undergo reproductive activation when housed with males. Finally, females allowed physical contact by being housed directly underneath males had heavier uteri than did females whose housing allowed contact only with the chemical cues from males. Among female arvicoline rodents, it appears that there exists a physiological continuum between absolute dependence on both contact and chemical cues from males vs. absolute independence for reproductive activation. The present results place female pine voles closer to the former extreme than to the latter. PMID- 8722625 TI - Regulation of plasminogen activator in rat endometrial stromal cells: the role of prostaglandin E2. AB - The rat endometrium undergoes decidualization, a tissue remodeling process, during embryo implantation. Plasminogen activator (PA), particularly the urokinase-type PA (uPA), has been implicated in tissue remodeling. The present study determined whether rat endometrial stromal cells secrete uPA during decidualization in vitro and, if so, whether the secretion is regulated by prostaglandins that are required in decidualization. Endometrial stromal cells were obtained from rats that had been treated with estrogen and progesterone to sensitize their uteri for decidualization, and the cells were cultured for up to 72 h in a serum-free medium. PA activity in the conditioned medium, as determined by a chromogenic assay, increased steadily during the 72-h culture period. PA secretion decreased when endogenous prostaglandin synthesis was inhibited by the addition of indomethacin to the cell cultures. The inhibitory effect of indomethacin on PA secretion was reversed by prostaglandin E2, and much less effectively by prostaglandin F2 alpha. PA activity in the medium was due primarily to uPA because 1) PA activity was inhibited by a uPA-specific inhibitor amiloride-and by an anti-mouse uPA antibody, and 2) the predominant PA activity in the medium, as identified in zymography, had a molecular mass of approximately 40 kDa, similar to that reported for uPA. Northern blot analyses of RNA from the cultured cells demonstrated that the steady-state levels of mRNA for uPA, but not for tPA and plasminogen activator inhibitor-1, were decreased by indomethacin; this decrease was reversed by prostaglandin E2. Taken together, the data indicate that rat endometrial stromal cells secrete uPA during decidualization in vitro, and that prostaglandin E2 regulates uPA secretion by the decidualizing cells by directly increasing uPA gene transcription and/or stabilizing its transcripts. These findings may help to partially elucidate the mechanism of action of prostaglandin E2 in decidualization. PMID- 8722627 TI - Development into blastocysts of bovine oocytes cryopreserved by ultra-rapid cooling. AB - The objective of the research described was to devise an efficient procedure to cryopreserve in vitro-matured bovine oocytes, using in vitro fertilization (IVF) and development of resultant zygotes into blastocysts as criteria of oocyte survival. Oocytes at metaphase II were found to be extremely sensitive to chilling. Cooling them to O degrees C for as little as 5 sec significantly decreased their capability to cleave and develop further after IVF; after 80 sec at 0 degrees C, only approximately 10% of chilled oocytes developed into blastocysts. Oocytes were also adversely affected by brief exposures to 4 M and 5.5 M ethylene glycol (EG) solutions supplemented with sucrose; after being suspended in either of these EG solutions in plastic straws and plunged directly into liquid nitrogen (LN2), few of the oocytes were fertilized and developed. To "outrace" chilling injury, oocytes contained in < 1 microliter of EG solution were placed onto electron microscope grids and plunged directly into N2 slush or LN2. After such ultra-rapidly cooled oocytes were warmed, 30% of them cleaved after IVF, and half of these developed into blastocysts-- survival rates equivalent to those for oocytes that had been exposed to EG without any cooling. This method offers promise as a novel way to cryopreserve bovine oocytes. PMID- 8722626 TI - Regulation of urokinase plasminogen activator production in implanting mouse embryo: effect of embryo interaction with extracellular matrix. AB - Embryo implantation in the mouse is an invasive process and requires the action of proteinases, including plasminogen activator (PA) and metalloproteinases. After the implanting embryo establishes close contact with the endometrium, the invasion process begins, at least in part, through interactions of the embryo with the extracellular matrix in the endometrium. This study determined whether embryo interaction with extracellular matrix components would affect the secretion of PA in vitro. PA in vitro. Mouse embryos were collected from the uterus on Day 3.5 of development, just before implantation, and were cultured dishes precoated with bovine serum, plasma fibronectin, or BSA (control). Embryos cultured on serum- or fibronectin-coated dishes secretes significantly more PA than those cultured on BSA. The effect of fibronectin was inhibited by hexapeptides that contained the integrin-recognizing Arg-Gly-Asp sequence. This indicates that the action of fibronectin in enhancing PA secretion is mediated through its receptor (integrins) in the embryo. Fibronectin fragments reproduced the effect of the whole fibronectin molecule, suggesting that the clustering of integrins by specific ligands is responsible, at least in part, for the increase PA secretion. The increase in PA secretion was a specific response to fibronectin rather than a reflection of increased total protein secretion, and was at least partially a result of the increased steady-state level of PA mRNA in the cultured embryos. Laminin was as effective as fibronectin in promoting PA secretion. Epidermal growth factor increased PA secretion, probably by promoting the interaction of the embryos with the extracellular matrix. In summary, our findings indicate that the interactions of the implanting embryos with their extracellular matrix may regulate trophoblast invasion by controlling PA secretion. PMID- 8722628 TI - Stress response of the rat testis: in situ hydridization and immunohistochemical analysis of heme oxygenase-1 (HSP32) induction by hyperthermia. AB - By using in situ hybridization and immunohistochemistry, the distribution patterns of heme oxygenase (HO)-1 (HSP32) transcript and protein were studied, and their response to thermal stress was examined. And, by using an HO-1 cDNA probe and polyclonal antibody, the levels of HO-1 mRNA and protein in normal and heat-shocked testis were quantified. The digoxigenin-labeled probe detected a strong signal for HO-1 transcript in Leydig cells, and in the Sertoli cells, spermatogonia, primary spermatocytes, and spermatids of the seminiferous tubules. In all cell types, the transcript was predominantly concentrated in the nucleus in a defined pattern. Thermal stress (42 degrees C, 20 min) did not change the cell population pattern of HO-1 transcript expression; however, it did cause distortion of the nuclear pattern and diffusion of the transcript signal in cells. Hyperthermic treatment of rats resulted in a modest (2- to 2.8-fold), time dependent, and sustained (1-16 h) increased in testicular 1.8-kb HO-1 mRNA. Immunohistochemical analysis of normal and heat shock patterns of testicular HO-1 expression showed robust staining of Sertoli ad leydig cells after heat shock; in normal tissue, immunoreactivity was low in these cell populations. As with the transcript distribution, hyperthermia did not affect the pattern of HO-1 immunoreactivity, and the protein was not detected in spermatogenic cells under control or stress conditions. In the Leydig cells, hyperthermia led to a more than 3-fold increase in the intensity of cytoplasmic staining for HO-1 protein. Consistent with the selective expression of HO-1, the level of the single HO-1 immunoreactive protein (approximately 32 kDa) detected in total testis microsomes showed a modest (1.5-fold) increase 6 h after heat shock. Data are consistent with te suggestion that differential distribution of HO-1 protein in the germ cell line and Sertoli cells reflects differential HO-1 mRNA processing in these cell types. The increase may be essential for the catalysis of the heme moiety of denatured hemoproteins such as cytochrome P450 and hemoglobin heme and hence may protect against heme-catalyzed free radical formation. We propose that induction of HO-1 protein in Sertoli and Leydig cells may function to protect the spermatogenic cells under conditions of thermal stress. PMID- 8722629 TI - Ovine endometrial expression of transforming growth factor beta isoforms during the peri-implantation period. AB - During the estrous cycle and early pregnancy, the uterus undergoes a variety o morphological changes. Because of their powerful effects on angiogenesis, on extracellular matrix modification, and on cellular proliferation and differentiation, the transforming growth factor beta (TGF beta) family of polypeptides may be involved in the regulation of pregnancy-related endometrial modification. In this study, endometrial stead-state mRNA expression of the three isoforms, TGF beta 1, beta 2, and beta 3, was quantified, and the proteins were localized during the later part of the estrous cycle (Day 13 and 16) and during early pregnancy (Days 13 through 30) in sheep. TGF beta 1 mRNA was expressed as a single transcript with steady-state mRNA expression levels 2-fold higher on Day 16 of the estrous cycle than on Day 13 of the estrous cycle (p < or = 0.002) or Days 13 and 16 (p < or = 0.004) and 0.008, respectively) of gestation. During pregnancy, levels increased progressively to peak on Day 27 (2.5-fold above Day 16 of pregnancy; p < or = 0.0001) and then leveled off at Day 30. Immunocytochemical localization of TGF beta 1 demonstrated protein in glandular protein in glandular and luminal epithelium at all days examined. TGF beta 2 mRNA was expressed as five distinct transcripts, and mRNA expression levels were lowest in Day 16 pregnant endometrium. Thereafter the expression level increased steadily through Day 30 (p < or = 0.0003). TGF beta 2 protein was localized in epithelium, diffusely within the endometrial stroma, and in leukocyte-like cells within the stroma. TGF beta 3 was expressed as one major transcripts and two minor transcripts. As with TGF beta 1, there was a dramatic difference in TGF beta 3 levels between Day 16 of pregnancy and Day 16 of the estrous cycle (3.8 fold, p < or = 0.0001). In pregnant ewes, endometrial TGF beta 3 levels increased 1.9-fold (p = 0.13) between Days 16 and 23 and remained relatively constant through Day 30. Immunohistochemistry localized TGF beta 3 protein most prominantly in the subepithelial stroma of the caruncule from Day 16 of the cycle in endometrial tissue. The observed changes in mRNA and protein expression patterns of TGF beta s within the ovine endometrium suggest that TGF beta s play a role in restructuring and modifying endometrium for a subsequent estrous cycle and/or pregnancy. PMID- 8722630 TI - Embryo infiltration by maternal macrophages is associated with selective expression of proto-oncogenes in a murine model of spontaneous abortion. AB - The causes and precise mechanisms leading to early embryo loss in mammals remain largely unknown, especially from a molecular point of view. Using the CBA/J x DBA/2 murine model of early spontaneous embryo loss (25-30% embryo loss), we have previously demonstrated the involvement of infiltrating activated macrophages and their cytolytic products such as nitric oxide and tumor necrosis factor alpha (TNF alpha) in the etiology of early embryo loss. On the other hand, far fewer of the CBA/J x Balb/c conceptuses (5-10% embryo loss) displayed significant cellular infiltration and nitric oxide and TNF alpha. Having used probes for cellular activation markers, we now present evidence indicating that significantly increased expression of AP-1 family members, Ha-ras, Ki-ras, v-erbA, v-raf, v abl, and c-myc was present in 24.4% of the CBA/J x DBA/2 embryonic units that also harbored significant Mac-1, F4/80, and class II major histocompatibility complex (MHC) molecule cellular infiltration. In contrast, only 7% of CBA/J x Balb/c conceptuses displayed increased proto-oncogene expression and increased cellular infiltration. Therefore, macrophage infiltration, cellular activation as identified by the increased expression of proto-oncogenes, and the production of cytotoxic macrophage products are closely linked to early embryo loss. These data add to the evidence that activated maternal macrophages may be directly responsible for spontaneous pregnancy failure. PMID- 8722631 TI - Regulation of steady-state concentrations of messenger ribonucleic acid encoding prostaglandin F2 alpha receptor in ovine corpus luteum. AB - To investigate the regulation of ovine luteal receptors for prostaglandin F2 alpha (PGF2 alpha), reverse transcription-polymerase chain reaction was used to produce a 284-bp partial cDNA that was 98% identical to that reported for the bovine PGF2 alpha receptor (PGF2 alpha-R). In situ hybridization localized mRNA for PGF2 alpha-R specifically to large luteal cells. In experiment 1, pools of luteal tissue (n = 4/day) collected from ewes on Days 3, 6, 9, 12, and 15 of the estrous cycle were analyzed for mRNA encoding PGF2 alpha-R. There was no difference in mean steady-state concentrations of mRNA encoding PGF2 alpha-R among any of the days studied (range = 2.3 +/- 0.3 to 3.5 +/- 0.7 fmol PGF2 alpha R mRNA/ microgram poly[A]+ RNA as assessed by slot-blot hybridization). In experiment 2, ewes on Day 11 or Day 12 of the estrous cycle were administered PGF2 alpha, and corpora lutea were collected 4, 12, or 24 h later (n = 4-5 per time point). Nontreated (n = 4) or saline-treated (n = 4) ewes served as controls. Luteal concentrations of mRNA encoding PGF2 alpha-R were decreased (p < 0.05) at 4, 12, and 24 h after injection of PGF2 alpha. In experiment 3, ewes (midluteal phase) were administered saline, PGF2 alpha, phorbol 12-myristate 13 acetate (PMA), or LH via ovarian arterial injection, and luteal tissue was collected 0, 4, 12, or 24 h later (n = 3-4 per treatment per time). Steady-state concentrations of mRNA encoding PGF2 alpha-R were decreased (p < 0.05) by PGF2 alpha and PMA treatment (4 and 12 h) but were increased (p < 0.05) at 24 h after LH treatment. In summary, 1) mRNA encoding PGF2 alpha-R was localized to large luteal cells; 2) concentrations of mRNA encoding PGF2 alpha-R did not vary during the estrous cycle; 3) treatment with PGF2 alpha or PMA to activate protein kinase C decreased concentrations of PGF2 alpha-R mRNA within 4 h of treatment; and 4) administration LH increased concentrations of mRNA encoding PGF2 alpha-R 24 h following injection. PMID- 8722632 TI - Maternal cells are widely distributed in murine fetuses in utero. AB - Passage of maternal cells into conceptuses in utero is recognized but poorly defined in species with hemochorial placentation. Despite the potential importance for such a phenomenon in vertical disease transmission, only limited data address the frequency of material to fetal cell trafficking or the developmental stage of its initiation. A murine model system, involving transfer of LacZ-, scid/scid, or wild type (+/+) blastocysts to pseudo-pregnant, LacZ+ transgenic ROSA26 females provided both flow cytometric and in situ information. In 100% of the late-gestation pregnancies studied, nucleated LacZ+ maternal cells crossed to conceptuses. In 90% of scid/scid fetuses, nucleated maternal cells were present in at least one lymphoid organ and often in more than one organ. Thymus was the most frequent site for maternal cell detection while the highest proportions of maternal cells were found in liver. Maternal cells were also visualized in fetal lung, heart, and bone marrow. Maternal cell trafficking into scid/scid fetuses commenced about midgestation, coincident with maturation of a placental circulation. In late-gestation +/+ fetuses, maternal cells were found extensively throughout bone marrow but not in other organs. The presence of maternal cells within primary lymphoid organs of fetuses may influence the repertoire of the developing fetal immune system and may be an underappreciated mechanism for vertical disease transmission. PMID- 8722633 TI - Region-specific localization of retinoic acid receptor-alpha expression in the rat epididymis. AB - Vitamin A has been shown to be involved in spermatozoal maturation in the epididymis. The action of vitamin A in male reproduction is postulated to be mediated at least partly by retinoic acid receptor-alpha (RAR alpha). The objective of this study was to determine whether the RAR alpha gene exhibits regional specificity in its pattern of expression along the length of the epididymis. The results would indicate where in the epididymis vitamin A may be required during maturation of spermatozoa. Northern blot analyses of RNA from the epididymis revealed two major transcripts, 3.4 kb and 2.7 kb, similar to the two major transcripts found in testis. In situ hybridization analyses demonstrated the expression of transcripts in the luminal epithelia to be highest in the proximal caput, low in the corpus, and high again in the distal cauda. This regional specificity in expression of the RAR alpha transcripts along the epididymis was virtually identical to that for the proteins, as visualized by reactions with anti-RAR alpha antibody and immunohistochemical stains. These results suggest roles for RAR alpha in the regions of epididymal epithelium that are postulated to participate in spermatozoal maturation and storage. PMID- 8722634 TI - Prolactin-induced regression of the rat corpus luteum: expression of monocyte chemoattractant protein-1 and invasion of macrophages. AB - Monocyte chemoattractant protein-1 (MCP-1) is a potential mediator of the recruitment of monocytes/macrophages into the regressing corpus luteum (CL). We investigated whether the luteolytic effect of prolactin in the rat is associated with the expression of MCP-1 and an invasion of monocytes/macrophages. Ovulation was induced in immature female rats by injection of eCG (5 IU, s.c.) at 30 days of age. All rats were hypophysectomized 3 days later. Rats received injections of ovine prolactin (250 micrograms, s.c.) at 12-h intervals on Day 9, 10, and 11 posthypophysectomy; controls received injection of vehicle. Rats were killed by decapitation 24, 48, or 72 h after the first injection of prolactin or vehicle. In rats treated with prolactin, immunoreactive MCP-1 was detected in the CL at 24 h after the first injection, and a consistent level of staining was reached by 72 h with immunodetectable MCP-1 diffused throughout individual CL. The number of monocytes/macrophages in the CL (mean +/- SEM) increased significantly after prolactin treatment, from 3.1 +/- 1.8 at 24 h to 49.3 +/- 8.2 at 72 h (p < 0.05), and the number of monocytes/macrophages was different from that in control, vehicle-treated rats at 72 h (10.3 +/- 4.1; p < 24 and 72 h in prolactin-treated rats (p < 0.05). It is concluded that a potentially important component of the luteolytic effect of prolactin in the rat is the expression of MCP-1 and invasion of monocytes/macrophages into the CL. PMID- 8722635 TI - Effects of nitric oxide-related agents on opioid regulation of rat testicular steroidogenesis. AB - These studies examined whether nitric oxide (NO) mediates opioid suppression of testicular steroidogenesis. Adult male rats were treated with various combinations of a NO synthase (NOS) inhibitor (NG-nitro-L-arginine methyl ester; NAME), a NO donor (isosorbide dinitrate; ISDN), an opioid agonist (morphine, and an opioid antagonist (naltrexone). Serum LH and testosterone and testicular interstitial fluid (TIF) testosterone concentrations were then measured. Inhibition of NO production by NAME reversed morphine-suppressed testosterone secretion; treatment with the NO donor, ISDN, reversed naltrexone-stimulated testosterone secretion. NAME did not alter morphine's effects on LH secretion and attenuated morphine's suppression of hCG-stimulated testosterone secretion, indicating that these effects occur directly in the testes and are not dependent on LH secretion. Even though these effects suggested possible interactions between NO and opioid systems, no additive or synergistic effects were found with suppressive combinations of morphine and ISDN, or with stimulatory combinations of naltrexone and NAME at does that had little effect on testosterone secretion when given alone. These results indicate that opioid and NO exert independent effects on testicular steroidogenesis through separate pathways or mechanisms and that NO does not mediate opioid-induced testicular suppression. PMID- 8722636 TI - Microtubular dynamics in granulosa cells of periovulatory follicles and granulosa derived (large) lutein cells of sheep: relationships to the steroidogenic folliculo-luteal shift and functional luteolysis. AB - Indirect immunofluorescence light microscopy was used to monitor temporal perturbations in the microtubular (tubulin) system of granulosa/lutein cells in paraffin-embedded sections of periovulatory follicles and corpora lutea of sheep. Estrogen-active granulosa cells of preovulatory follicles not yet exposed to the gonadotropin surge immunostained intensely for tubulin. Immunostaining of the microtubular matrix diminished after the onset of the surge and coincident with an abrupt fall in follicular estradiol production. A transient period of microtubular retraction was characterized by low-level steroid hormone output. Microtubules reappeared with the approach of ovulation and increase in follicular progesterone biosynthesis (luteinization). Treatment of animals during the preovulatory period with colchicine, a drug that binds specifically with tubulin and interferes with microtubular assembly, obstructed the follicular shift toward progesterone. Microtubular dynamics (polymerization<->depolymerization) evidently underpin fundamental mechanisms of follicular steroidogenesis. Finally, corpora lutea were isolated from ewes on Day 10 of the estrous cycle before (0 h) and after administration of prostaglandin (PG) F2 alpha. There was a small augmentation in luteal concentrations of progesterone at 2 h, followed by a sharp decrease from 4 to 16 h. Luteal weights were reduced (structural regression) at 24 h. Sections of large (PG-sensitive) steroidogenic cells of control corpora lutea typically displayed a radiating microtubular network. After administration of PGF2 alpha, tubular matrices of large cells were scant; mitochondrial clustering was evident in transmission electron micrographs. Affixed disassembly of the cytoskeleton of large luteal cells may be a heretofore unrecognized event in the biomechanics of functional luteolysis--perhaps uncoupling cholesterol translocation to mitochondrial cytochrome P450 side-chain cleavage. PMID- 8722637 TI - Chromatin reorganization in rat spermatids during the disappearance of testis specific histone, H1t, and the appearance of transition proteins TP1 and TP2. AB - Transition proteins replace testis-specific histones and are finally replaced by protamines in the nucleus of germ cells during spermiogenesis. In this study, immunoperoxidase and immunogold localization were used to determine both qualitatively and quantitatively the intracellular distribution of testis specific histone (H1t), transition protein 1(TP1), and transition protein 2 (TP2) during rat spermatogenesis. H1t labeling was concentrated over heterochromatin in the nucleus of late-pachytene spermatocytes and spermatids up to mid-steps 10. In step 9 spermatids, H1t was confined to the caudal end of the nucleus where heterochromatin was still present, while in early step 10 spermatids, only a few of the nuclei remained caudally labeled. In late step 10 spermatids, a fibrillar chromatin network was distributed throughout the nucleus coincident with the loss of H1t. A statistically significant rise in TP1 and TP2 labeling density over control values was first encountered in the nucleus of step 11 spermatids coincident with the initiation of condensation of the fibrillar chromatin. The TP1 and TP2 labeling density progressively increased in nucleus of step 11-13 spermatids with the apical to caudal condensation of the fibrillar chromatin, In step 13 spermatids, the chromatin was homogeneously condensed throughout the nucleus. In the case of TP1, the nuclear labeling density gradually declined after step 13 and disappeared by step 17. In the case of TP2, the nuclear labeling density disappeared by step 16. This study shows that, coincident with the loss of H1t, the chromatin of the spermatid is reorganized into a fibrillar network, whereas, coincident with the appearance and progressive increase of TP1 and TP2, the fibrillar chromatin condenses in an apical to caudal direction in the nucleus of the spermatid. Thus the remodeling of chromatin structure during spermiogenesis appears to be a two-step process that is sequentially influenced by the loss of spermatid-specific histones and the appearance of transition proteins. PMID- 8722638 TI - Granulocyte macrophage colony stimulating factor induces Fc epsilon RII/CD23 expression on normal human polymorphonuclear neutrophils. AB - Three major molecules have been recognized as IgE-binding structures on hematopoietic cells: the heterotrimeric high-affinity receptor for IgE (Fc epsilon RI), the low-affinity receptor for IgE (Fc epsilon RII/CD23) and the Mac 2/IgE-binding protein (epsilon BP). The latter has been shown to be expressed on polymorphonuclear neutrophils (PMN), where it regulates IgE-dependent activation. Experiments were undertaken to determine whether the IgE-binding capacity of PMN is mediated exclusively by this molecule. No detectable binding of human myeloma IgE to unstimulated PMN from normal volunteers could be evidenced. In contrast, PMN stimulated with granulocyte macrophage colony stimulating factor (GM-CSF) (500 U/ml) for 24 h displayed positive IgE binding. This binding was significantly inhibited in the presence of mAb directed against Mac-2/epsilon BP and also in the presence of anti-CD23 mAb, but not of anti-Fc epsilon RI mAb or isotype-matched controls. By flow cytometry, CD23 expression was detected on GM CSF-primed PMN by several anti-CD23 mAb, including EBVCS-5, BB10 or Mab135, which recognize different epitopes. CD23 was also evidenced by immunocytochemistry in GM-CSF-primed PMN. By in situ hybridization, GM-CSF-treated PMN exhibited a hybridization signal for CD23 mRNA and the presence of the CD23b isoform-specific mRNA was detected by RT-PCR. These findings indicate that PMN can synthesize CD23 molecules under GM-CSF induction. This strong CD23 expression might be of physiopathological relevance in IgE-dependent activation during allergic processes. PMID- 8722640 TI - Researchers in psychiatry in Canada. PMID- 8722639 TI - The soluble form of the IL-6 receptor (sIL-6R alpha) is a potent growth factor for AIDS-associated Kaposi's sarcoma (KS) cells; the soluble form of gp130 is antagonistic for sIL-6R alpha-induced AIDS-KS cell growth. AB - Kaposi's sarcoma (KS) is most frequently associated with HIV-infected individuals (AIDS-KS). While AIDS-KS-derived spindle cells (AIDS-KS cells) contribute to the development of KS lesions, growth regulation of these cells in vivo is poorly understood. AIDS-KS cells express considerable amounts of the signal transducing subunit (gp130) of the IL-6 receptor, but only a scanty amount of its binding subunit (IL-6R alpha). This phenotype can account for the lack of IL-6 responsiveness of AIDS-KS cells. We now report that the soluble form of IL-6R alpha (sIL-6R alpha), lacking transmembrane and cytoplasmic regions, functions as a potent growth factor for AIDS-KS cells by making these cells responsive to IL 6. After exposure to sIL-6R alpha together with IL-6 in culture, AIDS-KS cells assumed a spindle-shaped morphology and showed a remarkable augmentation of growth, while IL-6 alone did not induce AIDS-KS cell growth. Even without the addition of IL-6, sIL-6R alpha induced significant growth levels of AIDS-KS cells. Since AIDS-KS cells express high levels of IL-6, it is likely that, in the presence of sIL-6R alpha, these cells acquire an IL-6 autocrine growth loop. Anti gp130 antibodies blocked the action of sIL-6R alpha on AIDS-KS cells; hence, we refer to sIL-6R alpha as a gp130-related AIDS-KS cell growth factor. In contrast, the soluble form of gp130 (sgp130) had inhibitory effects on AIDS-KS cell growth, thereby suggesting that a complex regulatory system is involved in the modulation of the gp130-mediated AIDS-KS cell growth. In recent years, soluble forms of IL 6R alpha and gp130 have been detected in the sera of healthy individuals and increased levels of sIL-6R alpha as well as IL-6 have been noted in the sera of HIV-infected patients. It seems reasonable to assume that perturbed production of sIL-6R alpha and sgp130 may play a crucial role in the development and regression of AIDS-KS lesions by directly acting on growth of KS cells through the gp130 mediated pathway. PMID- 8722641 TI - "While you're up, get me a grant": a guide to grant writing. AB - Over the past few years, the number of well-trained researchers has increased, but the amount of money available for research has remained the same or even decreased. Consequently, competition for support is keener than it ever was. How a grant is written may tip the balance between getting funded or not. This article provides some guidelines for writing better proposals, touching on the various parts of the protocol and how it should be packaged. It addresses both the overt purpose of the proposal, which is telling the reviewers what you want to do, why, and how; and the covert message, which is convincing them that you have the ability to do what you propose. The article also discusses the review mechanism, who it is who reads the grant, and why this matters. PMID- 8722642 TI - Research training and productivity among faculty: the Canadian Association of Professors of Psychiatry and the Canadian Psychiatric Association Survey. AB - OBJECTIVE: This survey assesses the research training and productivity of academic faculty in Canadian departments of psychiatry and compares the findings with those of colleagues in the United States. METHOD: A questionnaire was adapted to suit the Canadian milieu and was distributed to a target population of 2484, including a core 522 full-time faculty. RESULTS: The response rate among full-time faculty was 65%, but only 26.5% for clinical and adjunct faculty. A small proportion (16%) of our MD and a greater proportion (57%) of our PhD respondents were included in a fairly lenient definition of researcher. Departments seek to recruit PhDs with an active involvement in research. Overall there appear to be more similarities than differences in research interests with our colleagues in the United States. The pharmaceutical industry was the most frequently mentioned source of research funding for MDs, while the availability of a mentor was perceived as the most influential factor determining the choice of a research career. CONCLUSIONS: Recommendations include adequate exposure to research during medical school and residency as well as appropriate inducements for the recruitment and retention of practising researchers. PMID- 8722643 TI - Physician resource variables and their impact on the future pool of research expertise among psychiatrists: the Canadian Association of Professors of Psychiatry and Canadian Psychiatric Association Survey. AB - OBJECTIVE: To assess the impact of the increased proportion of female psychiatrists and trainees, the rapid reduction of international medical graduates allowed in the country, and the expanding number of practitioners with subspecialization on the future pool of psychiatrist researchers in Canadian academic departments of psychiatry. METHOD: A questionnaire was adapted to suit the Canadian milieu and distributed to a target population of 2484, including a core of 522 full-time faculty. RESULTS: Female psychiatrists responding to the questionnaire reported less research involvement overall than their male counterparts. International medical graduates with both undergraduate and residency training abroad reported more research interests than other graduate categories. Respondents training in a subspecialty showed no difference in research commitment. CONCLUSIONS: Concerted efforts must be made to recruit, support, and retain female researchers. Particular attention must be paid to developing research expertise among Canadian medical graduates. The trend towards subspecialization influences only the selection of research topic. PMID- 8722644 TI - Psychiatric practice for the coming decade. AB - OBJECTIVE: To identify important trends and themes that will affect psychiatric training and practice. METHOD: Selective literature review and reflections by the authors. RESULTS: Three principal themes are elucidated relating to empiricism and integration, accountability and collaboration, and training and the public trust. CONCLUSIONS: There must be debate and action on these and other themes to maintain the relevance of psychiatry to its changing context. PMID- 8722645 TI - Clozapine: current status and role in the pharmacotherapy of schizophrenia. AB - OBJECTIVE: This study evaluates clozapine and its present role in the pharmacotherapy of schizophrenia. METHOD: Clozapine's current clinical status is reviewed, as is its position with respect to other treatment options. RESULTS: Clozapine represents the prototype of "atypical" neuroleptics, with evidence of clinical efficacy in both positive and negative symptoms, as well as a diminished risk of extrapyramidal side effects. It is the only neuroleptic to date that has established itself as having little, if any, risk of tardive dyskinesia. More recent research has focused on its potential for overall savings in health care costs, as well as possible benefits in the area of neuropsychological functioning. CONCLUSION: Evidence suggesting that the course of schizophrenia can be altered by effective treatment favours a systematic approach that optimizes treatment options. While clozapine does not represent a 1st-line agent because of its risk of agranulocytosis, it has an integral role to play in treatment resistant schizophrenia or in individuals experiencing intolerable side effects with conventional neuroleptics. PMID- 8722646 TI - Competency assessments: perceptions at follow-up. AB - OBJECTIVES: To report on the perceptions of assessment of competency and its consequences on a group of clients and significant others at follow-up. METHODS: Ninety-five interviews were conducted using a carefully developed semistructured telephone interview of 24 clients and 71 family/caregivers, representing the perceptions of about 80 clients. RESULTS: There was general satisfaction in the competency assessment process. There was a perception that interests and rights were protected. Clients were seen to be less involved in all spheres of decision making regardless of capacity outcome. Clients and families were satisfied with how decisions were made. CONCLUSIONS: Follow-up study of competency assessment does not support the conclusions previously drawn based on court record studies that assessments are deleterious and frequently result in violations of rights. PMID- 8722647 TI - Characteristics of psychogeriatric patient visits to a general hospital emergency room. AB - OBJECTIVE: This study describes psychogeriatric patients who visit the Emergency Room (ER) in a teaching hospital with a comprehensive psychogeriatric service. METHOD: A survey assessing demographic and clinical characteristics was completed for every psychiatric ER patient during a 2-year period. RESULTS: One hundred and seventy-three ER patients aged over 60 (mean age 71.2) were assessed. Fifty-three percent were previously known to the psychogeriatric service. The majority of these previously known psychogeriatric patients who used the ER suffered from mood disorders. The majority of new psychogeriatric patients seen in the ER had an organic brain syndrome (OBS). CONCLUSIONS: Few OBS patients who visited the ER were previously known to the psychogeriatric service, but many new OBS patients accessed the psychogeriatric service via the ER. For patients with mood disorders, particularly if suicidal, the ER was used as part of their service delivery system. PMID- 8722648 TI - [Capgras syndrome: evolution of the hypotheses]. AB - OBJECTIVE: to trace the evolution of hypotheses concerning Capgras' syndrome. METHODS: The data consist of slightly over 60 studies published between 1866 and 1994 which were selected in terms of their innovative nature and relevance to the clinical description of the syndrome and to psychodynamic, neurological and neuropsychological interpretations. RESULTS: Two partially overlapping major stages can be identified in the evolution of hypotheses regarding the mechanisms of the syndrome. The 1st, beginning in 1923, is characterized by the predominance of psychodynamic interpretations. The 2nd, resulting from the observation of organic dysfunctions in a high percentage of cases, is distinguished by the advent of neurological interpretations, and by a few mixed hypotheses. CONCLUSIONS: Overall, this review highlights the broad diversity of viewpoints concerning the syndrome. It will be used as a basis for the following study, which is designed to show that it is possible to test each of the viewpoints experimentally. PMID- 8722649 TI - Re: Treatment of SAD. PMID- 8722650 TI - Geriatric endogenous cortisol psychosis--role of cortisol antagonists. PMID- 8722651 TI - [Anxiety and the defibrillator: the Foreign Legion]. PMID- 8722652 TI - Treating depression among alcoholics. PMID- 8722653 TI - Freud, Prozac, and the human condition. PMID- 8722654 TI - Acute psychosis caused by mefloquine prophylaxis? PMID- 8722655 TI - Severe depression with risperidone-induced EPS in an elderly schizoaffective patient. PMID- 8722656 TI - Mitochondrial disorders in neuro-ophthalmology. AB - The pathogenesis of mitochondrial disorders relevant to neuro-ophthalmology has been further clarified by identifying and correlating to disease manifestations the three primary pathogenic mutations of Leber's hereditary optic neuropathy, and by correlating genetic, biochemical, and histochemical abnormalities in the mitochondrial myopathies and encephalomyopathies. PMID- 8722657 TI - The optic nerve. AB - Recent articles in the scientific literature have described major advances in our understanding of the anatomy and vascular relationships of the optic nerve (cranial nerve II) and of the diagnosis and treatment of a variety of disorders affecting this nerve, including congenital anomalies of the optic disc, dominant hereditary optic neuropathy, anterior and retrobulbar arteritic and nonarteritic ischemic optic neuropathy, optic neuritis, Cuban epidemic optic neuropathy, toxic and nutritional optic neuropathies, radiation-induced optic neuropathy, AIDS related optic neuropathy, optic neuropathies caused by tumors, and papilledema. PMID- 8722658 TI - The retina. AB - Prominent amongst last year's diverse papers on the retina were a study of the clinical manifestations of dominant cerebellar ataxia with pigmentary macular dystrophy, a review of the pathogenesis of carcinoma associated retinopathy, the Ischaemic Optic Neuropathy Decompression Trial, and a review of congenital optic disc anomalies. Ocular complications of several neurosurgical procedures were also reported during this period. PMID- 8722659 TI - The ocular motor nerves. AB - Recent articles in the scientific literature have described major advances in our understanding of the anatomy and vascular relationships of the three ocular motor nerves (cranial nerves III, IV, and VI) and of the diagnosis and treatment of a variety of pathologic processes that damage these nerves, including ischemia, inflammation, and compression. PMID- 8722660 TI - Eye movements. AB - Recent discoveries emanating from basic electrophysiological, anatomical, and pharmacological studies have implications for the diagnosis and treatment of clinical disorders of eye movements caused by disease affecting the brainstem and cerebellum. Electrophysiological studies have better defined the role of the superior colliculus in programming of saccades, and the nucleus reticularis tegmenti pontis in saccade-vergence interaction. Pharmacological inactivation studies have demonstrated the important role played by gamma-aminobutyric acid in the normal control of gaze and lead to trials of new agents in the treatments of nystagmus. PMID- 8722661 TI - Vestibular and auditory disorders. AB - Benign positional vertigo, the most common cause of vertigo, can now be cured with a simple bedside maneuver. A series of recent publications have clarified the pathophysiology of benign positional vertigo and documented the efficacy of particle repositioning maneuvers for treating the condition. The number of genetic loci and abnormal genes associated with inherited auditory and vestibular disorders is rapidly expanding. Within the past years, genes have been identified for Usher syndrome, Alport syndrome, deafness with fixation of the stapes and familial periodic ataxia. These discoveries should lead to improved diagnosis and, eventually, specific treatments for these disorders. PMID- 8722662 TI - Functional neuroimaging: radiology. AB - Rapid advances, in respect of the visual and auditory systems, have occurred in neuroimaging over the past year in the area of so-called 'functional imaging', and in particular of functional magnetic resonance imaging. This has been accelerated by the introduction of echo-planar imaging, which allows for the very fast acquisition (< 100 ms/image) of multislice images. Radiology is no longer just a question of interpreting anatomy, for, with the ability to combine structure with function, we are now able to extract physiological information. This opens up exciting possibilities for advances in the field of neuroscience, and functional imaging will therefore form the main thrust of this review. PMID- 8722663 TI - The many windows of therapeutic opportunities in stroke. PMID- 8722664 TI - Primary and secondary stroke prevention. AB - Recent clinical trials have demonstrated that we can prevent stroke with appropriate medical and surgical therapy. Treatment of stroke risk factors, antiplatelet therapy, anticoagulant therapy, and carotid endarterectomy have all proven to be effective if applied in appropriate clinical circumstances. This review will highlight the areas in which we have clinical trial evidence to guide us, and will point out those circumstances in which uncertainty remains. PMID- 8722665 TI - Stroke in children and young adults. AB - This selective review of stroke in children and young adults addresses: (1) stroke in sickle cell disease, particularly the prognostic use of transcranial Doppler, (2) oral contraceptives as a stroke risk factor, (3) stroke in pregnancy, and (4) the prognosis of ischemic stroke in young adults. Factors relevant to the primary prevention and outcome of stroke are emphasized. PMID- 8722666 TI - Vascular neurosurgery: aneurysms, arteriovenous malformations, subarachnoid hemorrhage, and intracranial hemorrhage. AB - Cerebrovascular disease continues to be a major source of morbidity and mortality in our aging population. Endovascular therapy for the treatment of cerebral aneurysms and associated vasospasm is gaining credibility. Some arteriovenous malformations considered inoperable in the past can now be successfully treated using multimodality therapy. Quantitative measurement of cerebral blood flow is helping to identify patients at risk of ischemic events and defining new guidelines for intervention. New noninvasive imaging modalities may replace conventional angiography for surgical disease. Advances in technology continue to improve our ability to treat cerebrovascular disease prior to irreversible ischemic events. PMID- 8722667 TI - Endovascular treatment for acute and chronic brain ischemia. AB - Recent advances in intracranial vascular catheterization along with new microballoon technology have led to increasing interest in the use of interventional techniques to treat cerebrovascular disease. We review several areas in which interventional neuroradiology is emerging as an important component of a brain ischemia treatment program. PMID- 8722668 TI - Anti-adhesion molecule and nitric oxide protection strategies in ischemic stroke. AB - In this review we focus on recent efforts designed to elucidate the role of inflammatory cells and nitric oxide in promoting cerebral ischemic cell damage, the mechanisms by which these events exacerbate ischemic cell damage, and the associated therapeutic interventions to protect tissue from ischemic injury. PMID- 8722669 TI - Neuro-ophthalmology and neuro-otology. PMID- 8722670 TI - Cerebrovascular disease. PMID- 8722671 TI - Selected list of nursing books and journals. PMID- 8722672 TI - Unlicensed assistive personnel: a solution to dwindling health care resources or the precursor to the apocalypse of registered nursing? PMID- 8722673 TI - Clinical practice guidelines as a new source of middle-range theory: focus on acute pain. PMID- 8722674 TI - Oregon's physician-assisted suicide vote: its effect on palliative care. PMID- 8722675 TI - Is caring a moral trap? PMID- 8722676 TI - Strategic planning for international nursing education. AB - International educational program planning is a complex process that benefits from the use of a strategic planning framework. The NANC project is presented as a demonstration model, utilizing the concepts and principles of strategic planning as outlined in the Regis Model. Developing and following a strategic plan in international nursing education is a necessity, rather than a luxury, in order to provide the necessary direction for your future providing a suggested "Blueprint for Action." Employing an effective strategic plan as your individualized "Blueprint for Action" can mean the difference between success and failure in your international "bridge-building efforts." The extensive assessment, planning, and review process identified through the utilization of the Regis Model that preceded the implementation of the NANC project was the key factor to project success. It is hoped that utilization of a strategic planning model such as the Regis Model, used in this project, will provide a general "Blueprint for Success" for those who are committed to quality international nursing education. PMID- 8722677 TI - Akrasia, health behavior, relapse, and reversal theory. AB - Philosophical explanations of the phenomenon of akrasia have been used in this article to characterize some of the difficulties involved in successful behavior change. The theory of psychological reversals provides a framework for understanding various types of akratic behavior. In addition, empirical research on the temptations of persons trying to quit smoking has supported hypotheses generated by the theory. The theory suggests that resisting the temptation to engage in akratic behavior is a complex phenomenon. Different strategies are needed in different types of metamotivational states. Reversal theory provides a coherent system for understanding and predicting akratic behavior. It suggests possible interventions that would improve the probability of circumventing akrasia and that would improve behavior change efforts of nurses engaged in health-promotion and disease-prevention activities. PMID- 8722678 TI - Tuberculosis: what nurses need to know to help control the epidemic. PMID- 8722679 TI - Doctor-nurse game. PMID- 8722680 TI - Myelodysplastic syndromes. AB - These two issues of the Seminars in Hematology will provide the physician the necessary knowledge to help make sense of this somewhat confusing array of diseases. The subdivisions of MDS reflect the precision of our techniques of dissection, with morphological and histochemical analyses forming the foundation to identify and subdivide MDS. Although steady refinement has occurred over the last half-century, the basic morphologic technique is unchanged. Cytogenetic analysis, which has been possible since the 1960s and 1970s, should be done at least at initial presentation in all patients to provide refinement of diagnosis and prognosis. FISH is not, at this time, useful as a screening technique. Although the 1990s is an era of rapidly growing knowledge and technical abilities in molecular biology, the use of these techniques in MDS is in its infancy. Very few genes have been identified which are altered in MDS, although many must exist. The molecular assays continue to be cumbersome and impractical to use in the clinical laboratory and remain the domain of the research scientist. Nevertheless, in the future, molecular biology will enable the internist to give each individual a clearer diagnosis and prognosis and may even provide targetted therapies of patients with MDS. At this time the center of management is good supportive care. Some patients, however, will benefit from special interventions, which include the use of growth factors, BMT, and in selected patients, aggressive chemotherapy. Induction of differentiation of the abnormal hematopoietic clone remains only a dream, although some of the differentiation agents may have applicability for their ability to induce apoptosis and prevent growth of the MDS clone of cells. Many of the major advances in our knowledge of cancer developed through the study of hematopoietic malignancy. A lot of these advances are due to the ease of obtaining the abnormal cells. MDS provides an excellent model for studying the progression of cells from their normal to preneoplastic and fully transformed states. A lucid understanding of this progression can form the paradigm for basic science to study neoplastic progression, and the molecular biology techniques used for these studies will be the basic tools used by hematologists and oncologists in the future. PMID- 8722681 TI - Morphology and classification of the myelodysplastic syndromes and their pathologic variants. PMID- 8722682 TI - Recurring diagnostic problems in the pathology of the myelodysplastic syndromes. PMID- 8722683 TI - Cytogenetics of myelodysplastic syndromes. PMID- 8722684 TI - Molecular genetic aspects of myelodysplastic syndromes. PMID- 8722685 TI - Immunological abnormalities in myelodysplastic syndromes. PMID- 8722686 TI - Biologic and clinical implications of marrow culture studies in the myelodysplastic syndromes. PMID- 8722688 TI - Testis-specific expression of mRNAs for a unique human type 1 hexokinase lacking the porin-binding domain. AB - Several enzymes in the glycolytic pathway are reported to have spermatogenic cell specific isozymes. We reported recently the cloning of cDNAs representing three unique type 1 hexokinase mRNAs (mHk1-sa, mHk1-sb, and mHk1-sc) present only in mouse spermatogenic cells and the patterns of expression of these mRNAs (Mori et al., 1993: Biol Reprod 49:191-203). The mRNAs contain a spermatogenic cell specific sequence, but lack the sequence for the porin-binding domain that somatic cell hexokinases use to bind to a pore-forming protein in the outer mitochondrial membrane. We now report the cloning of cDNAs representing three unique human type 1 hexokinase mRNAs (hHK1-ta, hHK1-tb, and hHK1-tc) expressed in testis, but not detected by Northern analysis in other human tissues. These mRNAs also contain a testis-specific sequence not present in somatic cell type 1 hexokinase, but lack the sequence for the porin-binding domain. The hHK1-tb and hHK1-tc mRNAs each contain an additional unique sequence. The testis-specific sequence of the human mRNAs is similar to the spermatogenic cell-specific sequence of the mouse mRNAs. Furthermore, Northern analysis of RNA from mouse, hamster, guinea pig, rabbit, ram, human, and rat demonstrated expression of type 1 hexokinase mRNAs lacking the porin-binding domain in the testes of these mammals. These results suggest that hexokinase may have unique structural or functional features in spermatogenic cells and support a model proposed by others for hexokinase gene evolution in mammals. PMID- 8722687 TI - Germ cell-specific DNA and RNA binding proteins p48/52 are expressed at specific stages of male germ cell development and are present in the chromatoid body. AB - Proteins homologous to the Xenopus oocyte mRNA binding proteins mRNP3+4 and designated p48/52 have been identified in male mouse germ cells (1993: Dev Biol 158:90-100). Western and North-western blots of extracts from testes and isolated germ cells indicate that p48/52 are present during meiosis but reach their highest levels postmeiotically at a time when many mRNAs are stored. Here we analyze the cellular and subcellular distribution of p48/52 in rat and mouse testes by LM and EM immunocytochemistry using an anti-mRNP3+4 antibody. Immunolabeling was found to be predominantly cytoplasmic and specific to germ cells at certain periods during their development. p48/52 were first detected in early pachytene spermatocytes at stage V of the seminiferous cycle and progressively increased during the remainder of meiotic prophase to a post meiotic peak in steps 1-8 round spermatids; thereafter, labeling gradually declined as elongated spermatids underwent nuclear condensation and elongation. A proportionally higher concentration of cytoplasmic immunolabeling was found within the lacunae of the anastomotic granulofilamentous network of the chromatoid body. The pattern of synthesis of these mRNA binding proteins together with their association with the chromatoid body suggests a role as germ cell specific mRNA stabilizing and/or storage proteins. PMID- 8722689 TI - Insulin-like growth factor (IGF-I) mRNA and IGF-I receptor in trout testis and in isolated spermatogenic and Sertoli cells. AB - Few data exist concerning the occurrence and potential role of an insulin-like growth factor (IGF) system in fish gonads. Using Northern and slot blot hybridization with a specific salmon IGF-I cDNA, we confirmed that IGF-I transcription occurs in trout testis. Testicular IGF-I mRNA abundance may be increased by long-term GH treatment in juvenile fish, while shorter treatment with growth hormone (GH) or a gonadotropin (GTH-II) in maturing males had no statistically significant effect. Radiolabelled recombinant human IGF-I binds with high affinity to crude trout testis preparation, to cultured isolated testicular cells, and to a membrane fraction of these cells (Ka = 0.2 to 0.7 x 10(10) M-1; Bmax = 10 to 20 fmol/10(7) cells, and 68 fmol/mg protein of membrane). The binding site was identified as type 1 IGF receptor by its binding specificity (IGF-I > IGF-II >>> insulin) and the molecular size of its alpha subunit labelled with 125I-IGF-I (M(r)125-140 kDa). 125I-IGF-II also bound to the type 1 receptor whereas IGF-II/ mannose 6 phosphate receptors could not be detected. Separation of isolated testicular cells by Percoll gradient and centrifugal elutriation provided populations enriched in different types of intratubular cells. IGF-I mRNA (detected by reverse transcription + polymerase chain reaction [PCR]) and IGF-I receptors (measured by competitive binding) were observed to a greater extent in Sertoli cell-enriched populations and in spermatogonia with primary spermatocytes. Therefore, IGF-I is a potential paracrine/autocrine regulator inside the spermatogenic compartment and appears as a possible mediator of GH action at the gonadal level in fish. PMID- 8722690 TI - Cloning, expression, and localization of a new member of a Paracentrotus lividus cell surface multigene family. AB - We have isolated and characterized a cDNA clone corresponding to a new member of bep (butanol, extracted, proteins) Paracentrotus lividus multigene family coding for cell surface proteins. The cDNA, called bep3, encodes a 370 amino acid protein and shares the same structural organization in the coding region with other members of the same gene family already characterized. Expression of this clone studied by Northern blot and by whole mount hybridization shows that the bep3 messenger is transcribed during oogenesis and utilized till the gastrula stage, whereas at the prism stage, unlike other members of the same gene family, new synthesis of messenger occurs. By whole mount hybridization spatial distribution of bep3 messenger in egg and embryos is established. This messenger appears located in the animal half of the unfertilized egg and moves to the cortical zone after fertilization; it is not present in the structures derived by the vegetal part of the embryo, such as the micromeres of the 16-cell stage, the primary mesenchyme cells of the blastula, and the primary intestine of the gastrula. At the prism stage instead, hybridization of bep3 messenger is restricted to the part of the embryo that will give origin to the oral region as successively confirmed by hybridization at the pluteus stage. The result of whole mount hybridization was confirmed by Northern blot hybridization of separated meso-macromere and micromere RNAs. A Southern blot experiment demonstrates that bep3 is codified by a single copy gene. Conservation of the bep multigene family in several Mediterranean and Japanese sea urchin species has also been analyzed. PMID- 8722691 TI - Differential expression of laminin chain-specific mRNA transcripts during mouse preimplantation embryo development. AB - Laminin is the first extracellular matrix protein that has been shown to be synthesized in preimplantation mouse embryos. In the present study, chain specific expression patterns of laminin mRNAs were examined by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR). During preimplantation mouse embryo development, temporal expression patterns of laminin chain mRNAs were somewhat differential: B1 chain mRNA was first detectable at the late two cell stage and its level was gradually increased by the blastocyst stage. In contrast, B2 and A chain mRNAs first appeared at the morula and blastocyst stages, respectively. At the blastocyst stage, all of the laminin chain mRNAs were highly detected compared to the earlier stages. When embryos were flushed at the morula stage and cultured in vitro, all laminin chain mRNA levels were decreased or not changed in the process of blastocoele expansion. In contrast, in the in vivo condition where embryos at different stages of blastocyst were flushed at different time points, laminin chain mRNA levels were increased as a function of blastocoele expansion. These changes in laminin mRNAs were parallel with its receptors such as integrin alpha 3 and alpha 6. 3-Isobutyl-1 methylxanthine (IBMX), which is known to be a potent activator of blastocoele expansion and regulates cAMP metabolism, upregulated laminin expression (except B1 chain) in blastocysts cultured in vitro. In vitro cultured embryos normally developed up to the late blastocyst, although their development was delayed compared with the in vivo condition where laminin gene expression was gradually increased as the blastocoele expanded. These results indicate that laminin expression may not be involved directly in the regulation of blastocoele expansion. The uterine environment enclosing the preimplantation embryos appears, therefore, to play an important role in the regulation of laminin gene expression during blastocyst development. PMID- 8722692 TI - YAC transgenesis in farm animals: rescue of albinism in rabbits. AB - The generation of transgenic mice with mammalian genes cloned in yeast artificial chromosomes (YACs) has generated great interest in the field of gene transfer into livestock. Many of the problems associated with standard transgenesis-such as lack of crucial regulator elements and position effects related to the integration site, which lead to variation in expression levels irrespective of the dose of the transgene-have been practically overcome. The large size of YAC derived gene constructs (in excess of 1 Mb) facilitates the presence and transfer of all elements required for the faithful regulation of a gene. With the experiments discussed in this report, we have addressed the possibility of applying the obvious advantages of YAC transgenesis to farm animals. We have generated transgenic rabbits carrying a 250 kb YAC covering the mouse tyrosinase gene by pronuclear microinjection, and thus rescued the albino phenotype of the transgenic individuals. To date, this is the first demonstration of a successful transfer of large genetic units into the germ line of farm animals. This development might improve the occurrence of transgene expression at physiological levels and specific sites in livestock. YAC transgenesis therefore will be applied in genetic engineering, for example, in the production of pharmacologically interesting proteins encoded by large gene units and generating transgenic donors for xenotransplantation. PMID- 8722693 TI - Nucleologenesis in the cleaving bovine embryo: immunocytochemical aspects. AB - In vivo nucleologenesis was studied in bovine embryos by electron microscopic immunogold labelling of DNA, RNA, protein C23 and protein B23. We have used the classification of Kopecny et al. (1989b) and Kopecny (1990) dividing nucleologenesis in four steps: compact nucleolar precursor body (NPB), monovacuolated NPB, NPB containing secondary vacuoles and fully reticulated nucleolus. These different features of early bovine embryo nucleologenesis were mainly observed during the eight-cell stage. In the first step of nucleolar development, the association of compact NPB with DNA structures was observed. DNA was also labelled in some small secondary vacuoles appearing during the third developmental step. From the second step onward, the labelling of protein C23 was observed in the compact fibrillar network of the NPB. Protein B23 started to be labelled in the compact fibrillar mass at the third step. RNA labelling was also observed for the first time in NPB containing secondary vacuoles. Labelled RNA was located in the peripheral region of compact fibrillar mass as well as along the border of the vacuoles. In the reticulated nucleolus, the dense fibrillar component was found to contain both proteins and RNA. PMID- 8722694 TI - IGF-I and insulin regulate glucose transport in mouse blastocysts via IGF-I receptor. AB - The roles of glucose deprivation, insulin, and insulin-like growth factor I (IGF I) in the regulation of glucose transport in the mouse blastocyst were examined. Glucose transport, measured by uptake of 3-0-methyl glucose (3-OMG), was increased by 19% (P < 0.01) in response to glucose deprivation. Both IGF-I and insulin stimulated uptake, but IGF-I was 1,000-fold more potent than insulin, increasing uptake by 51% at 1.7 pM (P < 0.001). These effects began to appear after 20 min of incubation with growth factors, and required the simultaneous presence of glucose. The relative potencies of insulin and IGF-I suggest that the actions of IGF-I and insulin were both mediated via the IGF-I receptor. The inactivity of a specific agonistic insulin receptor antibody (B10) confirms this and suggests that this action may be independent of signalling through IRS-1. Cycloheximide decreased growth factor-stimulated transport by about 40%, indicating that both protein synthesis and transporter recruitment from cytoplasmic stores are responsible for maximal stimulation. These characteristics are consistent with GLUT1-facilitated glucose uptake and suggest that GLUT1 is the regulatable transporter in mouse blastocysts. Stimulation of GLUT1 may be a ubiquitous feature of the autocrine/ paracrine activity of IGF-I in cell growth and proliferation. PMID- 8722695 TI - Experimental manipulations of compaction and their effects on the phosphorylation of uvomorulin. AB - Compaction of the eight-cell stage mouse embryo is a critical event in the generation of different cell types within the preimplantation embryo. Uvomorulin, a member of the cadherin family of cell adhesion molecules, is important during compaction and its phosphorylation increases early in the eight-cell stage, suggesting that this posttranslational modification may be important for compaction to proceed. We have assessed the importance of the phosphorylation of uvomorulin during compaction by preventing, reversing, or inducing adhesion prematurely. The only condition that affected the overall level of uvomorulin phosphorylation was the prevention of compaction through prolonged exposure of four-cell embryos to low Ca2+. This treatment reduced the level of uvomorulin phosphorylation in eight-cell embryos, and perturbed its localization to regions of cell-cell contact. Thus, whilst the phosphorylation of uvomorulin does not appear to regulate directly uvomorulin's adhesive function, it may be associated with the redistribution of uvomorulin during compaction. PMID- 8722696 TI - Differential preimplantation development of rhesus monkey embryos in serum supplemented media. AB - Several media, some augmented with amino acids, have been formulated recently, based on simplex optimization, to support the preimplantation development of mouse embryos. For the highly limited studies on preimplantation development of nonhuman primate embryos, a complex medium (CMRL-1066) has been employed. Our objective was to compare the developmental ability of rhesus monkey embryos in a simple medium containing amino acids, KSOM/AA, with the complex media used previously. Zygotes (99) were recovered following in vitro fertilization (IVF) from six monkeys, allocated to either CMRL or KSOM/AA both containing 10% fetal calf serum (FCS), and monitored daily until reaching the expanded or hatched blastocyst stage. The distribution of cells between the inner cell mass (ICM) and trophectoderm was determined at the end of culture by differential nuclear staining. Although a greater number of embryos cultured in KSOM/AA vs. CMRL developed to the morula stage (80%) and beyond (66% to expanded blastocyst), the differences were not significant. Such embryos in KSOM/AA did, however, develop at a significantly faster rate, on average, reaching the expanded blastocyst stage 26 hr earlier than did embryos cultured in CMRL. KSOM/AA embryos hatched in less time and had a higher percentage (43 vs. 34) of cells allocated to the ICM. These results indicate that a simple medium, KSOM/AA, in the presence of serum, supports the development of rhesus monkey embryos at high efficiency and at a faster rate than that observed for embryos cultured in the complex medium, CMRL 1066. PMID- 8722697 TI - Uterine lipid alterations during early pseudopregnancy and following the artificial induction of decidualization by Concanavalin A in QS mice. AB - Lipid extracts of whole uterine tissue from mice were examined by gas chromatography-mass spectrometry during days 2,3, and 4 pseudopregnancy (day 1 = copulatory plug) and following the artificial induction of the decidual cell reaction (DCR) on day 4. The range of lipids identified during pseudopregnancy and their percentage composition on day 2 included saturated fatty acids (SFA, 38%), monounsaturated fatty acids (MUFA, 20%), polyunsaturated fatty acids (PUFA, 17%), sterols (25%), long chain alcohols (0.12%), and alkylglycerols (0.11%). Of these, the main components were the fatty acids 16:0 (21%), 18:0 (14%), cis18:In 9 (14%), 18:2n-6 (8.5%), and cholesterol (24%). Although only subtle changes in the composition of uterine lipids occurred through days 2 and 3 of pseudopregnancy, more substantial changes were detected on day 4, at a time when the uterus normally initiates its transient "window of receptivity." Following induction of the DCR with the lectin Concanavalin A (Con A) at this time, even greater alterations in uterine lipid composition were observed. From 20 to 1,280 min post-Con A-treatment the percentage composition of SFA in the treated left uterine horn changed from 43% to 64%, sterols from 19% to 4%, PUFA from 15% to 10%, while MUFA remained unchanged at 23%. The lipid profile of the untreated right uterine horn of these animals was similar to that of the Con A-treated left uterine horn during the early stages. However, by 1,280 min substantial differences were observed, at a time corresponding with Con A-induced uterine growth. In contrast, differences in the lipid profile of Con A- and saline treated uteri were observed at 320 min post-treatment, a time preceding Con A induced uterine growth. Furthermore, the tissue concentration (nmol/mg dry weight) of SFA and sterols in uterine tissue decreased significantly following Con A treatment. The results suggest that uterine lipid changes are implicated in the development of uterine receptivity, and in the remodeling of uterine tissue for successful embryonic invasion and the establishment of pregnancy. PMID- 8722698 TI - Major secretory product of the mesometrial decidua in the rat, a variant of alpha 2-macroglobulin, binds insulin-like growth factor I via a protease-dependent mechanism. AB - Decidualization-associated protein (DAP), the quantitatively major secretory product of the mesometrial decidua in the rat, is a pl variant of the liver derived acute-phase reactant, alpha-2-macroglobulin (alpha 2M). Alpha 2M, a broad spectrum protease inhibitor, has been demonstrated in the human to bind a variety of cytokines and growth factors. In humans, the quantitatively major secretory product of decidual tissue is an insulin-like growth factor (IGF) binding protein. In this study, we have therefore tested the ability of liver- and decidual-derived alpha 2M in the rat to bind IGF-I. Alpha 2M purified from acute phase plasma and DAP purified from cytosolic extracts of decidual tissue and medium from tissue incubations both bound radiolabeled IGF-I. The binding of IGF I was principally dependent upon the coincubation of the protein with a proteinase. Therefore, it occurred during the conversion of the "slow" to the "fast" form of alpha 2M. Pretreatment with proteinase to produce the fast form before addition of the IGF-I reduced the binding. Binding was enhanced at a ratio protein:proteinase of 1:1. Results from gel electrophoretic analysis were consistent with the covalent linkage of IGF-I to alpha 2M during the cleavage of the "bait region." A saturable displacement by increasing concentrations of unlabeled IGF-I suggested high affinity interaction. Under conditions of demonstrated binding to purified proteins binding in acute-phase plasma, decidual tissue extracts and tissue incubation medium were associated with a high molecular weight species which was confirmed to represent alpha 2M and DAP, respectively. Our studies demonstrate that IGF-I may now be added to the list of regulatory peptides which alpha 2M may bind and that, in rat decidua, DAP may represent the functional homolog of decidual IGFBP-1 in the human and regulate growth factor function during placental development. PMID- 8722699 TI - Evidence for Ca(2+)-dependent ATPase activity, stimulated by decapacitation factor and calmodulin, in mouse sperm. AB - Membrane preparations from mouse sperm heads and tails were used in a gamma 32P ATP hydrolysis assay to investigate Ca(2+)-dependent ATPase activity. In membranes from sperm heads, but not tails, a Ca2(+)-dependent ATPase that was further stimulated by calmodulin was detected. The addition of partially purified mouse sperm decapacitation factor (DF) to head membrane preparations significantly stimulated Ca(2+)-ATPase activity, this effect being further increased in the presence of DF plus calmodulin; in contrast, no response was observed when the same treatment was applied to tail membranes. Sperm preincubated in the presence of trifluoperazine (TFP), a calmodulin antagonist, were significantly more fertile than cells from the same males incubated in the absence of TFP, indicating that inhibition of calmodulin accelerates capacitation. When sperm cells were preincubated briefly, then gently centrifuged to remove DF and resuspended in medium containing 45Ca2+ +/- DF, their ability to accumulate 45Ca2+ was significantly lower in the early stages after resuspension in the presence of DF than in its absence. These data correlated with chlortetracycline analysis of the sperm functional state. When cells were centrifuged and resuspended in medium only, there was a noticeable shift from the F pattern (characteristic of uncapacitated cells) to the B pattern (characteristic of capacitated cells), but the reintroduction of DF caused a significant reversion to the F pattern. Finally, using a monoclonal antibody to somatic cell Ca2(+)-ATPase, we have obtained evidence that the enzyme is particularly localized to the postacrosomal region of the mouse sperm head; specific binding was observed only in permeabilized cells, indicating that the epitope involved in the binding has an intracellular location. Based on these various pieces of evidence, we propose that when present on mouse sperm, DF stimulates calmodulin-sensitive Ca(2+)-ATPase activity and thus ensures maintenance of a low intracellular Ca2+ concentration. As capacitation proceeds, DF is lost and Ca2(+)-ATPase activity declines, allowing intracellular Ca2+ to rise and promoting capacitation-related changes. The fact that inhibitors of Ca(2+)-ATPase and calmodulin appear to accelerate capacitation in several mammalian species, as determined by chlortetracycline analysis, suggests that Ca(2+)-ATPase activity may play an important role in modulating capacitation in many or even all mammals. PMID- 8722701 TI - Morphological differences in nuclear materials released from hamster sperm heads at an early stage of incorporation into immature oocytes, mature oocytes, or fertilized eggs. AB - To elucidate the effects of ooplasmic factors on the early morphological changes in hamster sperm heads within the ooplasm, immature ovarian oocytes at the germinal vesicle stage (GV oocytes), ovulated fully mature oocytes, and fertilized eggs at anaphase II or the pronuclear stage (PN eggs) were examined in detail 15-30 min after insemination or reinsemination. Thin-sectioning studies demonstrated distinct materials released from the sperm nucleus over the entire postacrosomal nuclear surface immediately after disappearance of the sperm nuclear envelope. The release occurred in all of the oocytes and eggs prior to or even in the absence of subsequent chromatin decondensation. Depending upon the stage of the penetrated oocyte or egg, however, the materials varied in morphology: several hemispherical projections of amorphous material within mature oocytes; a number of electron-dense globules within GV oocytes and PN eggs; and both forms within eggs at anaphase II-telophase II. These observations and the fact that only the release of the amorphous material was accompanied by sperm chromatin decondensation indicate that this release was the initial process of chromatin decondensation, whereas the release of the globules resulted from a deficiency or lack of ooplasmic factors affecting the sperm nucleus. Restriction of the release in both forms of material to the late meiotic phase suggests changes in the factors associated with progression of meiosis. To approach an understanding of the mechanism of successful decondensation of sperm chromatin, the ooplasmic factors considered responsible for the stage-dependent release of nuclear materials are discussed. PMID- 8722700 TI - Glucose utilization during gonadotropin-induced meiotic maturation in cumulus cell-enclosed mouse oocytes. AB - Earlier work from this laboratory has determined that glucose plays an important role in the mechanisms regulating meiotic maturation in mammalian oocytes. In the current study, we have further explored the role of glucose in hormone-induced germinal vesicle breakdown (GVB) in an effort to better understand how glucose utilization and metabolism relate to the control of meiotic maturation in mouse cumulus cell-enclosed oocytes (CEO). When CEO were cultured in medium containing 4 mM hypoxanthine (to maintain meiotic arrest), 5.5 mM glucose, and 0.23 mM pyruvate, follicle-stimulating hormone (FSH) stimulated lactate accumulation in a time-dependent manner. Addition of 2-deoxyglucose (2-DG) to the medium at various times after the initiation of culture resulted in rapid termination of lactate production and suppression of FSH-induced GVB scored after 18 hr of culture, the effectiveness diminishing the longer the delay before addition of 2-DG. By 8 hr, addition of 2-DG was without effect on GVB. Similar effects were seen when FSH treated CEO were washed free of glucose. In a 2-DG dose-response experiment, gonadotropin-induced lactate production was prevented, but this inhibition did not necessarily prevent GVB. The activities of six metabolic enzymes were measured in extracts of freshly isolated complexes, and in order of increasing activity were: hexokinase, 6-phosphogluconate dehydrogenase, glucose-6-phosphate dehydrogenase, phosphofructokinase, lactate dehydrogenase, and pyruvate kinase. Of the six enzymes examined, only hexokinase activity was increased in CEO exposed to FSH. CEO were cultured in microdrops in the presence or absence of FSH, and aliquots from the same microdrop were assayed for glucose, lactate, and pyruvate. In response to FSH, utilization of glucose in microdrop cultures by CEO was markedly increased and was accompanied by comparable lactate production and limited pyruvate production. Cycloheximide and alpha-amanitin both blocked FSH induced oocyte maturation, but only cycloheximide prevented the increase in hexokinase activity and glucose consumption. These data suggest that hexokinase is an important rate-limiting enzyme for glucose utilization that is under translational control and participates in the mechanisms controlling the reinitiation of meiosis. However, stimulation of glycolytic activity does not appear to be a necessary concomitant for meiotic induction. PMID- 8722702 TI - Occurrence of androgen and estrogen receptor mRNAs in the harderian gland: a comparative survey. AB - In Rana esculenta the presence of an androgen receptor in both the male and female Harderian gland (HG) has been demonstrated. Hybridization analysis has evidenced a high degree of homology between the rat androgen receptor cDNA and the frog androgen receptor mRNA (fARmRNA). Correspondingly the molecular size of fARmRNA is similar to those described in mammals (9.4 kb). In in vivo experiments testosterone (T) increases the levels of fARmRNA. The use of the antiandrogen alone or in combination with T prevents the increase of fARmRNA. In the control animals a loss of fARmRNA has been observed. In primary cultures of HG cells, the steady-state levels of fARmRNA increase in the cells exposed to T. These results suggest that T exerts an autoinduction on its own receptor, increasing the levels of fARmRNA. In Xenopus laevis the HG shows a sexual dimorphism of the protein pattern. The female shows two major proteins (210 and 180 kDa). Administration of estradiol to the male shifts the protein pattern into the female one. In this respect an estrogen receptor mRNA (ERmRNA) has been found in the female gland and can be induced in the male one. No ARmRNA has been detected in either sexes. A similar sex dimorphism has been found in Gallus domesticus. The female pattern is characterised by a protein fraction of about 210 kDa, the male one by a protein fraction of about 180 kDa. In 4-day-old chicks no sex differences have been found. An ERmRNA is expressed in the female, while no ARmRNA has been detected in both sexes. Neither AR nor ER mRNAs have been detected in the chick HG. Among mammals the HG or the hamster (Mesocricetus auratus) shows an androgen-dependent sex dimorphism. In in vitro experiments T 10(-12) M induces a onefold increase of ARm-RNA with respect to unexposed cells. This effect reaches its maximum (4.4 fold) when cells are exposed to T 10(-8) M. The size of the hamster ARmRNA is similar to that observed in other mammals (9.5 kb). The above results suggest that in the HG the phenomenon of autoinduction occurs and that there is a relationship between the androgen or estrogen dependence of the HG and the digamety of the species. PMID- 8722703 TI - Lipid secretory mechanisms in the mammalian harderian gland. AB - The mammalian Harderian glands are lipid-secreting glands. In an unstimulated condition, the glandular cells frequently exocytose the lipid materials; however, no intracellular calcium ion ([Ca2+]c) changes are detectable. Cholinergic (muscarinic) secretagogues induce secretory activity and increase of [Ca2+]c. A G protein activator, sodium fluoride, enhances the secretory activity and increase of [Ca2+]c. Removal of extracellular calcium ions inhibits the secretion enhanced by cholinergic stimulation. Under pharmacologic stimulation, glandular cells may show an apocrine-like secretory pattern. Cholinergic stimulation also induces contraction of the myoepithelial cells covering glandular end pieces; however, the reduction in volume of glandular end pieces is not prominent. Catecholamines have no effect on the release of lipid materials. These results indicate the involvement of G-proteins linking with muscarinic receptors and Ca2+ dynamics (increase of [Ca2+]c and Ca2+ influx) in lipid secretion by glandular cells and in contraction of myoepithelial cells of mammalian Harderian glands. However, the increase of [Ca2+]c in Harderian glands was less when compared with other cells- for instance, those which secrete protein. PMID- 8722704 TI - Isolation and identification of sex-specific cDNA clones from the Syrian hamster harderian gland. AB - Syrian hamster Harderian glands show a typical sexual dimorphism, with males having two secretory cell types and females having one cell type and intraluminal porphyrin accretions, among other differences. Since these differences may be due to the expression of specific genes, our interest is to identify those genes and their role on the development and control of the sexual dimorphism. The experimental approach was to construct cDNA libraries for male and female Syrian hamster Harderian glands and then subtracted libraries for male vs. female and for female vs. male. By this method, cDNA libraries enriched either in male specific or in female-specific clones were obtained. Clones from those libraries were checked for differential expression by using double colony hybridization with [32P]-cDNA from male and female glands. Then, the selected clones were checked again for expression in Harderian glands by Northern hybridization, using poly(A+) RNA from males, castrated males, and females. Finally, the clones were sequenced and compared to search for significant homologies. One of the male specific clones showed strong homology with rat cytochrome p450b/e. Among the female-specific clones, homologies were found to the complement C3 fragment from several species, to sequences from the mouse mammary tumor virus, and to the subunits C1 and C2 of the rat prostatic steroid binding protein. Several other clones showed no significant homologies and need further characterization. PMID- 8722705 TI - Expression of the somatostatin gene and receptors in the rat harderian gland. AB - Somatostatin is one of the numerous peptides described in the Harderian gland of different animals. With the aim of trying to elucidate its physiological role, we investigated whether this peptide is expressed in OFA rat Harderian gland at different ages and seasons and, if so, studied the regulatory proteins involved in the activation of the somatostatin gene, and also whether it contains any somatostatin receptors. Nursing (4-15-day-old), prepubertal (21-30-day-old), and adult (54-day-old) OFA rats were sacrificed by decapitation throughout the year, and the Harderian glands were excised and immediately frozen in liquid N2. The expression of somatostatin and its receptors was investigated using RT-PCR techniques; additionally, the existence of proteins which bind to cAMP responsive elements (CRE) was investigated using a band-shift technique. The somatostatin gene was expressed in the Harderian gland of rats aged 4-30 days in autumn and winter but not in spring and summer or in older animals. However, the somatostatin receptor was expressed throughout the year at all the ages studied. In the autumn, nuclear proteins binding to CRE (CREB) were present in 8-10-day old rats but not in younger 4-day-old animals. We conclude that rat Harderian gland cells transcribe the somatostatin gene depending on the season and age of the animals, while its receptor is always present at all the ages studied; the CREB found produces the same retardation complex as ICER (inducible cAMP early repressor), an isoform of CREM (cAMP responsive element modulator), which in the pineal has been shown to be under adrenergic control. Since somatostatin expression is regulated by cAMP mechanisms, it is feasible that the existence of this repressor ICER could explain why somatostatin expression disappears in adult animals once maturation is complete. PMID- 8722706 TI - Hormones and the control of porphyrin biosynthesis and structure in the hamster harderian gland. AB - The hamster Harderian gland seems to present both an excellent model for the control of porphyrin biosynthesis and an unusually robust example of the interrelationship between structure and function. It has been known for some time that 1) the capacity for manufacturing and storing porphyrins and 2) gland histology and ultrastructure are controlled by androgens. Thus, in intact males as well as in gonadectomised animals of either sex treated with androgens, porphyrin synthesis by the Harderian gland is suppressed and the gland tubules characteristically possess two cell types, the cytoplasm of both containing polytubular complexes. By contrast, the Harderian glands of intact females and castrated males synthesise and store large amounts of protoporphyrin, while their tubules possess only one cell type which lacks a polytubular complexes. So overarching is the effect of androgens that they have been described as a "coarse tuning" effect on the gland. By contrast, the role of the ovary is both less dramatic and less well understood. In female hamsters, ovariectomy leads to degenerative changes in Harderian gland tubules and (probably) a release of stored porphyrin; at the same time there is a reduction in enzyme levels and new synthesis. The causative hormone in this "fine tuning" is unclear at present. There is now clear evidence that the Harderian gland is also controlled directly by pituitary hormones. In particular, the use of continuous infusion osmotic minipumps has allowed us to demonstrate not only 1) that the expected rise in porphyrins and feminisation of gland morphology does not occur in castrated males receiving the dopamine agonist bromocriptine, but that 2) the simultaneous administration of prolactin does permit these changes; furthermore, 3) the administration of prolactin alone increases porphyrin synthesis above the levels found in untreated castrates. Similarly, bromocriptine administration to ovariectomised females markedly reduces porphyrin synthesis and masculinises gland structure; again, this is reversed by the simultaneous administration of prolactin. Prolactin must therefore be seen as equipotent with androgens in determining gland structure and activity. PMID- 8722707 TI - Sexual dimorphism in the harderian gland of the Syrian hamster is controlled and maintained by hormones, despite seasonal fluctuations in hormone levels: functional implications. AB - The Harderian gland of the Syrian hamster (Mesocricetus auratus) is unusual amongst rodents in the degree of dimorphism present. Other types of hamsters have Harderian glands which are apparently identical in male and female animals. Laboratory populations of Syrian hamsters are derived from very limited genetic stock, which makes one concerned lest they not be representative of wild populations; however, until wild stocks of M. auratus become available, we should assume that insights derived from studies of dimorphism in Syrian hamsters represent important considerations for the life of these animals. Two dimorphic features are the histology and the porphyrin content of the Harderian glands. About 95% of the lipid droplets in female glands are small (type 1), whereas only about 65% of those in males in type 1, with the other 35% being type 2 (large droplets). Five weeks of castration of males led to an increase in type 1 droplets to 90%. On the other hand, 2 weeks treatment of females with testosterone led to a reduction in type 1 droplets to about 82%. Short day photoperiods led to a large increase in type 2 droplets in both males and females (to 52% in males, 35% in females after 8 weeks). These results suggest that the lipid contained in type 2 droplets is important to hamsters of both sexes during the winter. Porphyrin concentrations are 100-1,000 times higher in females than males, and this is largely controlled by testosterone as orchidectomy leads to increased male levels and testosterone treatment leads to reduced female levels. However, a number of treatments which also lead to reduced testosterone levels do not lead to increased porphyrins and may, in fact, prevent the rise which would normally follow orchidectomy. One of these antiporphyrinogenic treatments is exposure to short day photoperiods. Thus, the sexual differences in porphyrin, levels in Syrian hamsters are maintained, despite seasonal fluctuations in hormone levels. This suggests that this dimorphism is important for the function of the gland. PMID- 8722708 TI - Characterization of binding sites for beta-adrenergic agonists and vasoactive intestinal peptide in the rat harderian gland. AB - Vasoactive intestinal peptide (VIP) receptors and beta-adrenergic receptors were investigated in rat Harderian gland membranes using 125I-VIP and 125I cyanopindolol (125I-CYP), respectively, as ligands. The receptor bindings were rapid, reversible, saturable, specific, and dependent on time, temperature, and membrane concentration. The stoichiometric data suggested the presence of two classes of VIP receptors with Kd values of 0.36 and 65.37 nM and binding capacities of 323 and 39,537 fmol VIP/mg protein, respectively. The interaction showed a high degree of specificity, as suggested by competitive displacement experiments with several peptides structurally or not structurally related to VIP as follows: VIP > helodermin > rGRF > PHI > > secretin. Glucagon, somatostatin, insulin, and pancreastatin were ineffective at concentrations up to 1 microM. However, the stoichiometric data suggest the presence of one class of binding sites for 125I-CYP. The Kd for the single site was 290 pM with a binding capacity of 32 pmol/L. The pharmacological characterization of 125I-CYP binding to membranes showed that only isoproterenol, a beta-adrenergic agonist, and norepinephrine, an alpha beta-adrenergic agonist, was as effective as propranolol in inhibiting 125I-CYP binding to Harderian gland membranes. However, alpha 1- and alpha 2-adrenergic agonists and blockers such as methoxamine, prazosin, clonidine, and yohimbine were shown to be ineffective. These results demonstrate the presence of specific VIP and beta-adrenergic receptors in the Harderian gland and suggest a role for VIP and beta-adrenergic agonists in the physiology of this gland. PMID- 8722709 TI - Mg(2+)-dependent and Ca2+, Mg(2+)-dependent ATPase activities in the harderian gland of rodents: age and sex influences. AB - Three experiments employing male and female Syrian hamsters (aged 1, 2, and 8-10 months), male Sprague-Dawley rats (aged 1, 2, and 10 months) and male C57B1 mice (aged 2, 7, 13, and 29 months) examined the effects of age and sex on Mg(2+) dependent and Ca2+, Mg(2+)-dependent ATPase activity in the Harderian gland. Significant differences due to age and sex were observed in the hamsters and rats but not with age in mice. Generally, male hamsters had significantly higher Mg(2+)-dependent and Ca2+, Mg(2+)-dependent (exception at one timepoint) ATPase activity than did females. Age-matched male and female rats had similar values of Mg(2+)-dependent ATPase activity, but males had significantly higher Ca2+, Mg(2+) dependent ATPase activity than females at 2 months of age. PMID- 8722710 TI - Immune cell proliferation in the harderian gland: an avian model. AB - Experimentation has been carried out to study proliferation of plasma cells in the chicken Harderian gland (HG) and to determine if a HG factor influences immune cell (i.e., B cell) proliferation. In young chickens, flow cytometric analysis of propidium iodide (PI)-stained plasma cells revealed that the percentages of cells in both the synthetic (S) and mitotic (G2M) phases of the cell cycle were highest between 6 and 9 weeks of age. A pattern of plasma cell depletion and repopulation in the HG was observed following administration of emetine dihydrochloride. At 3 and 5 days posttreatment the plasma cell population decreased, and by 7 days posttreatment repopulation of the gland with plasma cells occurred. This repopulation appeared as a result of plasma cell proliferation within the HG. Anti-5-bromo-2'-deoxyuridine (BrdUrd) staining of frozen sections showed that the numbers of plasma cells incorporating BrdUrd were low at 3 days posttreatment but were as high, or higher than, controls at 5 and 7 days posttreatment. These results were verified with flow cytometric data of PI stained plasma cells. Data from bursal cell bioassays revealed proliferative activity influenced by a HG factor. Coculture of bursal cells with phorbol dibutyrate and diluted HG supernatants resulted in prolonged and increased proliferation of these cells. It is possible that the HG of chickens supports plasma cell proliferation through the elaboration of a factor which acts like a lymphokine. PMID- 8722711 TI - The harderian gland in autoimmune diabetes of the nonobese diabetic mouse. AB - Infiltration of the nonobese diabetic (NOD) mouse's Harderian gland (HG) was studied in 1-30-week-old animals. A mononuclear cell invasion of this gland is first seen in 8-week-old female mice (i.e., at a slightly later age than that for the onset of infiltration of pancreatic islets). Infiltrating elements are mainly located at the hilus of the gland or at one or two foci (periacinar infiltration) within the parenchyma. In the latter case, a few elements infiltrate the fibrous connective tissue surrounding the acini (one or more) without damaging them. The most severe histopathological lesion was observed in 16-week-old animals; at this time infiltration ranges from a still focal lesion to complete acinar destruction of the gland. Ultrastructural observations confirm that in several cases acinar cells are destroyed and the HG parenchyma is substituted with infiltrating elements, fibroblasts, and connective tissue. HG infiltration is comparable to the pancreatic inflammatory infiltration; the two processes are very similar, though insulitis starts slightly earlier than HG infiltration. Furthermore, as for insulitis and diabetes incidence, HG infiltration affects NOD males less than females. Moreover, immunocytochemistry has shown that T lymphocytes are the prevalent infiltrating element both in pancreatic islets and HGs. Further studies are required to understand the reasons for autoimmune destruction of this gland. PMID- 8722712 TI - The lymphoid substance of the chicken's harderian gland is organized in two histologically distinct compartments. AB - Light and electron microscopical investigations revealed that the lymphoid structure of the chicken Harderian gland is organized in different histological frameworks. In the head the surface epithelium of the central canal can be classified as a lymphoepithelial tissue which covers the dense lymphoid substance. It consists of small and medium-sized lymphocytes, dendritic-like cells, and occasional macrophages. High endothelial venules are associated with intense lymphocyte migration and homing that gives circumstantial evidence for a T-dependent region, as found in a secondary lymphoid organ. The B-dependent germinal centers are also common structural units of the head region's lymphoid substance. The body of the gland is loaded with plasma cells of different maturation stages. They immigrate into the epithelium of the central canal and produce IgM and IgA. Only a few scattered IgG producing plasma cells can be found in the gland of Harder. This plasmocytic region accounts for the immunosurveillance on the conjunctiva and in the upper respiratory tract through antibody production against bacterial or parasitic infections. In both the head and body regions of the gland, anti-B-L (anti-Ia) antibody recognized scattered elongated cells which might represent dendritic cells. The immunological relationship between the two histologically different parts of the Harderian gland is unknown, but we speculate that the dense lymphoid tissue with high endothelial venule receives the blood-borne, immunologically mature, but uncommitted B cells. By the influence of local antigen stimulus, these B cells transform to plasma cells which gradually appear in the body of the gland. The lymphoid structures of the head and the body fulfill the function of secondary and tertiary lymphoid organs, respectively. PMID- 8722713 TI - Monitoring water pathways through mungbean pods using backscattered electron microscopy. AB - Weather damage by rainfall is a major constraint to production of high quality mungbean seed. Breeding mungbeans for weathering resistance is assisted by an understanding of the pod wall structure and how this structure can affect water absorption. This paper describes a simple microscopic technique for determining the transport route of salt solutions from the external pod surface through the pod wall tissues and into the pod lumen. Different mungbean cultivars were selected based on differences in field performance. Mungbean pods were immersed in concentrated salt solutions (lanthanum nitrate, caesium chloride and sodium chloride) for different time periods, embedded in resin blocks and polished prior to backscatter imaging in a field emission scanning electron microscope. The salts precipitated between the pod wall tissues and through timed experiments clearly demonstrated the passage of salt solution through the pod wall tissues and into the pod lumen. The sale molecules penetrated the outer epidermis and parenchyma but were unable to penetrate the dense sclerenchymatous layer beneath. The salt solution entered the lumen of the pod via the small gap between the suture cap and the dense tissue sheath. Although this technique may not provide a true determination of fresh water absorption through mungbean pods it does demonstrate a simple means of identifying more resistant pod structures suitable for use in achieving genetic improvement. PMID- 8722714 TI - Meta-analysis of clinical studies of the efficacy of granulocyte transfusions in the treatment of bacterial sepsis. AB - BACKGROUND: Meta-analysis was used to explain disagreements across controlled clinical studies of the efficacy of granulocyte transfusions (GTX) in the treatment of bacterial sepsis. METHODS: Studies published in English in 1970-1994 were retrieved. Seven studies of adults and five of neonates were eligible for analysis. Summary relative odds (RR) of survival in treated patients versus controls were computed for patient subsets defined on the basis of microbiologic proof of infection, recovery of bone marrow function, method of procurement of granulocytes for transfusion, dose of granulocytes transfused, assessment of leukocyte compatibility, and survival rate of controls. The random-effects method was used for all analyses. RESULTS: Differences between the reviewed studies in the dose of granulocytes transfused and the survival rate of controls were primarily responsible for the disagreements across the reports. Adults (RR = 4.2) and neonates (RR = 18.0) receiving adequate doses of granulocytes and adults transfused in centers with a low survival rate of controls (RR = 8.9) experienced a significant (P < .05) benefit from GTX. CONCLUSION: GTX of adequate dose may be indicated in the 1990s for the treatment of sepsis in neonates, and perhaps also adults admitted to centers with an unusually high mortality rate of untransfused controls. More research is needed to reassess the proper role of GTX, in the light of modern transfusion medicine technology and the presently available options for the treatment of sepsis. PMID- 8722716 TI - In vitro study on the removal of pemphigus antibody with immunoadsorption. AB - The adsorbability of pemphigus vulgaris (PV) antibodies to immunoadsorbent with phenylalanine, tryptophan, and dextran sulfate as ligands was examined. PV antibodies were adsorbed to immunoadsorbents with both phenylalanine and tryptophan as ligands, in batchwise adsorption and column adsorption. PV antibodies were not adsorbed to immunoadsorbent with dextran sulfate as ligand, in column adsorption. It was also shown that the rates of decrease of normal protein fractions in column adsorption with phenylalanine as a ligand were small. Our results suggest that immunoadsorption therapy using sorbent with phenylalanine as a ligand may be useful for the treatment of PV. PMID- 8722715 TI - Plasmapheresis in primary dysfunction of hepatic transplants. AB - BACKGROUND: Primary dysfunction is a failure of graft function which occurs in approximately 5% of transplanted livers. Retransplantation is often required. The presence of elevated serum cytokines interleukin 6 and tumor necrosis factor with hepatic graft dysfunction, as well as the historical benefit of plasmapheresis in fulminant hepatic failure-associated coma suggest a possible role for plasmapheresis therapy in the management of primary graft dysfunction in liver transplantation. DESIGN AND METHODS: We evaluated the effectiveness of plasmapheresis in the management of primary graft dysfunction in 18 patients who underwent orthotopic liver transplantation in this institution. Patients who were diagnosed with primary dysfunction of hepatic grafts underwent a course of four daily plasma exchange procedures. The clinical outcome, patient and graft survival, was compared to that of historical controls. RESULTS: Graft survival at 10 days was 77.7% and 76.2% and patient survival at 100 days was 83.3% and 85.7% in the plasmapheresed and control groups, respectively. The patients who underwent plasmapheresis had a higher incidence of dialysis intervention, 38% versus 19%, indicating more severe graft dysfunction. In the small number of patients compared for concomitant dialysis therapy, patient survival in the plasmapheresed group was 85.7% versus 50% (control), and graft survival was 57.0% versus 50%. Serum cytokine levels of tumor necrosis factor and interleukin 6 were reduced by 66.0% and 55.2%, respectively, with a single procedure. CONCLUSION: Plasmapheresis did not significantly effect graft survival in patients with primary graft dysfunction. An increase in patient survival in severe graft dysfunction with renal failure was noted but was not significant. Removal of elevated serum cytokines TNF and IL-6 was documented. PMID- 8722717 TI - Anti-DNA antibody kinetics following selective removal by adsorption using dextran sulphate cellulose columns in patients with systemic lupus erythematosus. AB - The aim of this study is to determine by mathematical analysis which of two models, the one- or the two-compartment model, more closely approximates the kinetics of anti-dsDNA following immunoadsorption procedures in patients with systemic lupus erythematosus. Titers of anti-dsDNA were measured at specified intervals after apheresis to each model by nonlinear least-squares methods, and Akaike's Information Criterion (AIC) was calculated to determine which model most approximately described the kinetics. The AIC of the two-compartment model was larger than that of the one-compartment model in all 14 SLE patients (P < .001). Therefore, the one-compartment model is thought to be suitable. The generation rate and catabolic rate of anti-dsDNA were obtainable using this model. The anti dsDNA replenishment curve after an immunoadsorption session was defined by only two parameters: the generation and catabolic rates of anti-dsDNA. PMID- 8722718 TI - Determining factors for the outcome of peripheral blood progenitor cells harvests. AB - This is a pilot retrospective study to investigate the factors that may affect the collection of peripheral blood progenitor cells (PBPC). Sixty-nine PBPC harvests in 18 cancer patients (median age 39.5; 8 males and 10 females) were performed during marrow recovery after chemotherapy and hematopoietic growth factors. Median number of nucleated cells (MNC) collected were 13.3 (range 2.3 44.5) x 10(9) per session. Median CFU-GM was 362 colonies (range 63-1,720) per 500,000 MNC. Neither sex, body weight, diagnosis, nor the number of days into leukapheresis was significantly associated with MNC and CFU-GM. Older patients tend to have higher CFU-GM in the PBPC harvests (P = .0437). Higher WBC on the day of harvest is significantly associated with higher yield of MNC after leukapheresis (P < .0001). Patients without any evidence of disease have significantly higher yield of MNC than those having local/distant metastases with or without marrow involvement (P = .0302 and .0446). For patients with metastatic disease, those with bone marrow involvement tend to have higher CFU-GM than those without bone marrow involvement although the difference is not statistically significant (P = .0559). Those patients who have received only one, or three and more chemotherapy regimens have a higher yield of MNC than those who have only two previous chemotherapy regimens (P = .036 and .0324). The mechanism of PBPC mobilization is also discussed. In view of the limited patient number in this study, the results should be confirmed by larger studies. PMID- 8722719 TI - Providing leukocyte-reduced platelets using the CS-3000 PLUS. AB - Reducing leukocyte (WBC) contamination of platelet (PLT) concentrates diminishes some adverse effects associated with transfusions. To provide WBC-reduced PLTs, we initiated a program using bedside filtration. However, the inability to easily quantitate WBC removal and PLT loss at the bedside prompted us to perform filtration in the blood bank. To establish optimal methods, production of WBC reduced PLTs using the CS-3000 PLUS was studied in three phases, during which technical modifications were made. During phase 1, prestorage WBC reduction was performed using the PALL LRF-10H filter, sterilely connected. WBC reduction was satisfactory, but PLT loss was excessive. During phase 2, the PLT-30 collection chamber and Fenwal Closed System Apheresis Kit with Integral Sepacell Leukocyte Reduction Filter were used. PLT yields were improved, but now WBC contamination was excessive. During phase 3, the interface offset was reduced from 10 to 6, and both PLT yields and WBC reduction were satisfactory. Using this final method (CS 3000 PLUS, PLT-30 collection chamber, integral filter and offset setting of 6), the mean PLT yield per unit is 4.29 x 10(11) (N = 1,146), and the mean WBC contamination is 0.50 x 10(6) (N = 32). PMID- 8722720 TI - High-avidity anti-DNA antibody removal from the serum of systemic lupus erythematosus patients by adsorption using dextran sulfate cellulose columns. AB - The purpose of this study is to determine whether immunoadsorption treatment using a dextran sulfate (DS) column can remove high-avidity anti-double-stranded DNA (anti-dsDNA) antibody from the blood of patients with systemic lupus erythematosus (SLE). Before and after each immunoadsorption therapy routine, titers of the high-avidity anti-dsDNA antibody of 11 SLE patients were measured by using a newly developed assay kit to exclusively detect high avidity anti-DNA antibody. Patients with active SLE showed significantly higher titers of high avidity antibody than did those with inactive SLE, and their titers were significantly reduced by immunoadsorption procedures. Removal of high-avidity antibodies in vitro was also confirmed by mixing patients' sera and DS gel. Immunoadsorption therapy using DS columns is effective in the removal of high avidity anti-dsDNA antibodies that are closely associated with pathogenicity in SLE. PMID- 8722722 TI - Organizational guidelines for therapeutic apheresis facilities. American Society for Apheresis Standards and Education Committee. PMID- 8722721 TI - Reversal of acute renal allograft rejection by extracorporeal photopheresis: a case presentation and review of the literature. AB - There is a clear need for well-tolerated immunomodulatory agents that can aid in the prevention of acute solid organ rejection. Extracorporeal photopherosis is an apheresis-based therapy that is currently available at many medical centers worldwide. Preliminary studies utilizing photopheresis with standard immunosuppressives have shown this therapy to successfully reverse acute cellular rejection of cardiac allografts with minimal toxicity. No formal evaluation of the role of extracorporeal photopheresis had been performed in renal transplantation. In this report, photopheresis was successfully utilized to treat acute cellular rejection in a patient with a renal allograft. This lends further support to the existing literature suggesting that photopheresis may be useful for the reversal of acute solid organ rejection. Although our experience with this patient is anecdotal, photopheresis merits further study as treatment for severe renal allograft rejection. PMID- 8722723 TI - A new treatment of refractory ascites: ascitoapheresis. PMID- 8722724 TI - Symptomatic hyperviscosity syndrome with polyclonal gammopathy responding to therapeutic plasmapheresis in a patient with myelodysplastic syndrome. PMID- 8722725 TI - Plasmalymphacytapheresis in the treatment of cold autoimmune hemolytic anemia. PMID- 8722726 TI - A case of thrombocytopoenic thrombotic purpura preceding onset of systemic lupus erythematosus. PMID- 8722727 TI - Preemption and prevention. PMID- 8722728 TI - The pain medicine specialist as a physician-healer. PMID- 8722729 TI - Measuring the activity of older people with chronic pain. AB - OBJECTIVE: A variety of instruments have been applied to the measurement of activity, yet few, if any, have been validated specifically for older people with chronic pain. This study has sought to examine the utility of the Human Activity Profile (HAP) for describing activity in a sample drawn from a pain clinic for older people. DESIGN: The HAP was administered to 193 older pain clinic patients, 72 of whom completed the profile on a second occasion. A further 55 responses were collected from a group of community-dwelling volunteers. The factor structure of the HAP was tested using these 320 responses. The factors subsequently derived were compared with the Sickness Impact Profile (SIP) and the Barthel Index (BI). The discriminant validity of the HAP was examined by comparing factor scores for groups determined by gender, diagnosis, and status in the pain clinic. RESULTS: The 94 items of the HAP loaded onto 10 factors, which explained 63.7% of the variance. These factors demonstrated moderate associations with the BI and the subscales of the SIP. The factors discriminated between men and women (F[12.180] = 9.85. p < 0.000). Differences were also present between subjects with a musculoskeletal pain problem, postherpetic neuralgia, and pain free volunteers (F[24.340] = 4.7. p < 0.000). Factor scores increased between pre and postclinic assessments (F[12.60] = 4.79. p < 0.000). CONCLUSIONS: The HAP has demonstrated qualities which favor its adoption as an activity measure for older pain clinic patients. PMID- 8722730 TI - Premature Infant Pain Profile: development and initial validation. AB - OBJECTIVE: Inadequate assessment of pain in premature infants is a persistent clinical problem. The objective of this research was to develop and validate a measure for assessing pain in premature infants that could be used by both clinicians and researchers. DESIGN: The Premature Infant Pain Profile (PIPP) was developed and validated using a prospective and retrospective design. Indicators of pain were identified from clinical experts and the literature. Indicators were retrospectively tested using four existing data sets. PATIENTS AND SETTINGS: Infants of varying gestational ages undergoing different painful procedures from three different settings were utilized to develop and validate the measure. METHODS AND RESULTS: The largest data set (n = 124) was used to develop the PIPP. The development process included determining the factor structure of the data, developing indicators and indicator scales and establishing internal consistency. The remaining three data sets were utilized to establish beginning construct validity. CONCLUSIONS: The PIPP is a newly developed pain assessment measure for premature infants with beginning content and construct validity. The practicality and feasibility for using the PIPP in clinical practice will be determined in prospective research in the clinical setting. PMID- 8722731 TI - Gender differences in the expression of depressive symptoms among chronic pain patients. AB - OBJECTIVE: To investigate the extent to which gender differences in the expression of depressive symptomatology exist among chronic pain patients. SETTING: A multidisciplinary pain center affiliated with a state medical school in the southern United States. PATIENTS: A total general sample of 245 patients with heterogeneous chronic pain complaints randomly selected from a database of approximately 1,000 pretreatment evaluation patient files. A depressed subsample of 113 patients based on scores > or = 15 on the Beck Depression Inventory (BDI). MEASURE: The 21-item BDI. RESULTS AND CONCLUSIONS: Gender differences in total BDI scores were not revealed for the total general sample or depressed subsample, but gender differences in the expression of depressive symptoms (i.e., item level responses) were found via separate discriminant function analyses on the total general sample and depressed subsample. Consistent with previous studies performed on diverse samples, females' higher endorsement of body image distortion was noted in both of our samples. Females also endorsed significantly higher levels of fatigue in our total sample. Items that were not significant in our total sample include females' higher levels of loss of appetite and crying. In the depressed subsample, items probing pessimism and failure were not statistically significantly different to males' endorsing higher levels of each. Sensitivity to these gender differences is suggested in clinical practice. The likely consequences that these differences in the experience and expression of distress have on coping activities (e.g., help-seeking responses) and the reactions of others (e.g., health-care providers) are highlighted. PMID- 8722732 TI - Myofascial pain syndrome of the peroneus longus: biomechanical approach. AB - OBJECTIVE: The aim of the study was to investigate the role played by anatomical (lower limb length discrepancy) and biomechanical (alterations in the dynamics of movement) factors in the pathophysiology of myofascial pain syndrome (MPS) of the peroneus longus. DESIGN: Patients affected with MPS of the peroneus longus of one side were submitted to either correction of their lower limb discrepancy by heel lift (6 patients) or normalization of altered biomechanical parameters during movement, as measured via ground-foot reaction analysis (g-f) by dynamic insoles (6 patients) for 60 days. At days 7, 15, 30, and 60, the effect of treatment was verified on painful symptoms [VAS scale, presence of the active trigger point (TrP) in the muscle] and on g-f parameters [peaks of vertical force (F1 and F3 of Fz) and of lateral shear force (Fx)]. RESULTS: Treatment with heel lift produced a moderate, significant reduction of the spontaneous pain and of the abnormal Fx peak in the affected leg after 7 to 14 days with no further improvement afterward. Treatment with dynamic insoles caused a marked, significant reduction of the pain at 7 days, with complete resolution of the painful symptoms at 30 days and concomitant disappearance of the active TrP in the muscle. It also produced a significant and progressive reduction of the abnormal Fx peak in the affected leg starting at the 7th day and continuing until the 60th day. CONCLUSION: Both anatomical and biomechanical alterations of the dynamics of movement play a role in the painful symptoms of MPS of the peroneus longus, but the biomechanical factor is by far the more prominent. PMID- 8722733 TI - Changes in daily hassles, mood, and sleep in the 2 days before a migraine headache. AB - OBJECTIVE: The determination of the changes in the incidence and stressfulness of daily hassles as well as in the mood states: alert, tense, irritable, annoyed, depressed, and tired and in the quality of sleep in the 2 days before a migraine headache. DESIGN: Prospective cohort study. SETTING: Private headache practice. PATIENTS: Twenty female migraine patients. INTERVENTIONS: Diary four times per day for 10 consecutive weeks with the variables quantified through the use of 100 mm visual analogue scales. RESULTS: The incidence and stressfulness of daily hassles as well as the mood states: tense, irritable, annoyed, depressed, and tired were significantly increased in the 2 days before a migraine headache in comparison to control days. CONCLUSIONS: The results confirm previous studies that daily hassles are increased in the days before a migraine headache. They also show that the increase in daily hassles is associated with mood changes, particularly consisting of tension and depression. PMID- 8722734 TI - A psychoanalytic investigation to improve the success rate of spinal cord stimulation as a treatment for chronic failed back surgery syndrome. AB - OBJECTIVE: The analysis of patient data concerning psychological structure and functioning produced an instrument to determine whether a neurostimulator ought to be implanted or not. DESIGN: A questionnaire containing 24 items was developed by a psychologist and tested in 40 chronic failed back surgery patients for whom a spinal cord stimulation seemed to be the only therapeutic approach. This questionnaire was based upon some crucial psychological themes, on which the patient took a position. A predictive indication factor (I.F.; %) for implantation of the neurostimulator was obtained from the 24 items. Six months after the implantation of the neurostimulator, we correlated the evaluation factor (E.F.; %) with a six-point evaluation scale considering the pain reduction. The aim was to compare the I.F. and E.F. to verify the correlation between them. This comparison was intended to answer the question if psychological variables included in our scale improve the success rate of the therapy. SETTING: Data were collected by a psychologist at the Pain Clinic of the University Hospital of Gent, Belgium. RESULTS AND CONCLUSIONS: The correlation between the I.F. and the E.F. was calculated for the 40 patients by the Spearman correlation test. A coefficient value of 0.8083 (p = 0.000) was found, indicating the existence of a very close correlation between the predictive I.F. and the E.F. The indication scale appears to be a useful instrument for clinical psychologists to predict the success rate of a spinal cord stimulator in this group of patients. PMID- 8722735 TI - Acute pain after thoracic surgery predicts long-term post-thoracotomy pain. AB - OBJECTIVE: Long-term pain is a common sequela of thoracotomy, occurring in approximately 50% of patients 2 years after thoracic surgery. Despite this alarming statistic, little is known about the factors responsible for the transition of acute to chronic pain. The aim of the present study is to identify predictors of long-term post-thoracotomy pain. DESIGN: Follow-up was for 1.5 years for patients who had participated in a prospective, randomized, controlled trial of preemptive, multimodal analgesia. SETTING: Subjects were recruited from a tertiary care center. PATIENTS: Thirty patients who had undergone lateral thoracotomy were followed up by telephone, administered a structured interview, and classified according to long-term pain status. MAIN OUTCOME MEASURES: Present pain status was measured by a verbal rating scale (VAS). Measures obtained within the first 48 h after surgery were compared between patients with and without pain 1.5 years later. These include VAS pain scores at rest and after movement, McGill Pain Questionnaire data, patient-controlled morphine consumption (mg), and pain thresholds to pressure applied to a rib contralateral to the thoracotomy incision. RESULTS: Fifty-two percent of patients reported long-term pain. Early postoperative pain was the only factor that significantly predicted long-term pain. Pain intensity 24 h after surgery, at rest, and after movement was significantly greater among patients who developed long-term pain compared with pain-free patients. A significant predictive relationship was also found at 24 and 48 h using the McGill Pain Questionnaire. Cumulative morphine was comparable for the two groups. Pain thresholds to pressure applied to a rib contralateral to the incision did not differ significantly between the groups. CONCLUSION: Aggressive management of early postoperative pain may reduce the likelihood of long-term post-thoracotomy pain. PMID- 8722736 TI - Gabapentin adjunctive therapy in neuropathic pain states. AB - OBJECTIVE: This is a report of a trial of the new antiepileptic agent gabapentin in patients with intractable neuropathic pain. DESIGN: A case series of patients with a diagnosis of neuropathic pain whose previous management was inadequate were given oral gabapentin in increasing doses and were followed for a minimum of 2 months, monitored for efficacy and side effects. SETTING: An outpatient pain management center located within a major university medical center. PATIENTS: Convenience sample of patients referred for management of intractable neuropathic pain. INTERVENTIONS: Simplification of existing pharmacologic management, addition of gabapentin, and attempted reduction of opiate analgesic doses. MAIN OUTCOME MEASURES: Patient self-reports and pain scores in successive office visits. RESULTS: Gabapentin provides analgesic activity for patients with neuropathic pain and has the advantage of a low side effect profile and drug toxicity. PMID- 8722737 TI - Clonazepam for the treatment of lancinating phantom limb pain. AB - OBJECTIVE: To propose clonazepam for use in the empiric treatment of shooting/shocking phantom limb pain. SETTING: Outpatient pain clinic associated with a university hospital. PATIENTS: Two patients with phantom limb pain after total hip disarticulation. INTERVENTIONS: Treatment with clonazepam. RESULTS AND CONCLUSIONS: Clonazepam provided effective relief for > 6 months in two patients with shooting/shocking phantom limb pain. Although clonazepam therapy is not new, it appears to have been omitted from current pain texts and journals as a treatment option for phantom limb pain. PMID- 8722738 TI - Long-term intrathecal administration of midazolam and clonidine. AB - OBJECTIVE: To determine the clinical usefulness of the long-term intrathecal administration of midazolam and clonidine in patients with refractory neurogenic and musculoskeletal pain. SETTING: Pain Centre, Academic Hospital Groningen, Groningen, The Netherlands. PATIENTS: Four patients with chronic benign neurogenic and musculoskeletal pain, not responding to conventional analgesic therapy. OUTCOME-MEASURES: Visual Analogue Score, Activities of Daily Living. RESULTS: The intrathecal administration of midazolam and clonidine produced almost immediate and nearly complete pain relief. Even with continuous use, tolerance seemed to be no problem, and side effects appear to be minimal. CONCLUSION: Intrathecal infusion of midazolam and clonidine produced promising results in four patients with refractory chronic benign pain. Further research will be necessary to determine the efficacy and the risk-to-benefit ratio of long term administration of this combination. PMID- 8722739 TI - Psoas sheath chemical neurolysis for management of intractable leg pain from metastatic liposarcoma. AB - CASE REPORT: A 56-year-old man with widely metastatic liposarcoma, after left Tower extremity amputation, complained of severe right lower extremity pain. Trials of systemic opioids had resulted in poor pain control while introducing intolerable dose-limiting side effects. METHODS AND RESULTS: Initial inpatient management consisted of a lumbar epidural infusion of a dilute local anesthetic and preservative-free morphine. This provided satisfactory relief but was discontinued because of recrudescence of phantom limb pain. A lumbar epidural infusion of preservative-free morphine sulfate was associated with poor pain relief, central nervous system (CNS) side effects, and severe urinary retention resulting in acute renal failure. A repeated trial of parental opioids provided marginal pain relief with persistent CNS side effects. Chemical neurolysis of the lumbar plexus was performed with 10 ml of 10% aqueous phenol injected into the psoas muscle sheath. The pain gradually resolved over a 2-day period without apparent side effects. Motor function was preserved, pain was resolved, and as systemic opioids were reduced, cognitive function and overall well-being were improved. PMID- 8722740 TI - Assessing a new procedure: thoracic radiofrequency dorsal root ganglion lesions. PMID- 8722741 TI - Patient-controlled epidural analgesia during labor. PMID- 8722742 TI - Affected-sib-pair interval mapping and exclusion for complex genetic traits: sampling considerations. AB - We describe an extension of Risch's [(1990a,b) Am J Hum Genet 46:222-228, 229 241] method of linkage detection and exclusion for complex genetic traits. The method uses interval mapping to infer disease locus identity-by-descent (IBD) sharing for affected sib pairs (ASPs) based on marker information for the ASP and other genotyped family members. The method is likelihood based, and makes use of Risch's parameterization in terms of recurrence risk ratios for relatives. We describe specific linkage detection and exclusion tests for use as genome screening tools to prioritize genomic regions for further study. We also examine issues of optimal study design. We advocate initially typing a large panel of ASPs (and no additional family members) with a map of genetic markers evenly spaced at 10-20-cM intervals. We recommend a screening procedure that 1) investigates further all regions with maximum lod scores greater than 1 and 2) excludes from consideration those regions that result in lod scores less than -2 at the smallest genetic effect that is viewed as important to detect. Further investigation of an interval might include typing other available families or family members, typing additional markers in the interval, and carrying out further statistical analyses. This strategy is efficient in the number of genotypings required and focuses attention on regions most likely to harbor a disease gene with a substantial impact on disease risk, while resulting in the pursuit of a manageable number of false-positive linkage results. Modification may be required if insufficient ASPs are available or if families come from a significantly admixed population. PMID- 8722743 TI - Comparison of methods for survival analysis of dependent data. AB - Analysis of dependent survival data by conventional partial likelihood methods produces unbiased estimates of the regression coefficients but incorrectly estimates their variance. Here we compared the conventional partial likelihood methods with two alternative methods for analyzing dependent survival data. The first alternative method estimated the regression coefficient by the partial likelihood approach but adjusted the variance to account for clustering. The second alternative method used marginal likelihoods to estimate both the regression coefficient and its variance. We evaluated the performance of the three methods using simulated and actual data. Simulated data were used to examine bias, efficiency, type I errors, and power. An Old Order Amish genealogy was analyzed under these models to illustrate their performance on real data. The simulation study showed that all three methods provided unbiased estimates of the regression coefficient, but the efficiency of the estimated regression coefficient varied according to the simulation conditions. The standard partial likelihood method showed increasing type I error as the dependence increased within clusters. Both alternative methods had acceptable levels of type I errors at all dependence levels. In the analysis of genealogic data, the regression coefficient was similar in the three methods showing stable estimates of the regression coefficients. The variance estimates from the alternative methods were slightly different from the conventional method, suggesting a flow level of dependence. This study displays the effect of violating the independence assumption and provides guidelines for using alternative statistical methods. PMID- 8722744 TI - Influence of apolipoprotein E genotypes on plasma lipid and lipoprotein concentrations: results from a segregation analysis in pedigrees with molecularly defined familial hypercholesterolemia. AB - Familial hypercholesterolemia (FH) is a monogenic disorder caused by mutations in the low-density lipoprotein (LDL) receptor gene. Large variations in plasma lipids and lipoprotein levels have been observed in FH families. These may be caused by other environmental and genetic factors of which apolipoprotein E (apo E) is a candidate. The possible influence of apo E polymorphism on components of variation in plasma LDL-C, triglycerides, high-density lipoprotein cholesterol (HDL-C), and lipoprotein(a) (Lp(a)) levels was investigated in 235 members of 14 families with FH. Sex-and age-adjusted mean LDL-C was influenced significantly by the apo E genotype in non-FH subjects (P Ste20p-->Ste5p. Overexpression of Cdc42p resulted in a slight increase in pheromone induction of a reporter gene, and overexpression of activated forms of Cdc42p resulted in a further twofold increase. Mutations in pheromone response pathway components did not suppress the lethality associated with the activated CDC42 mutations, suggesting that this effect is independent of the pheromone response pathway. PMID- 8722767 TI - Genetic interactions between REG1/HEX2 and GLC7, the gene encoding the protein phosphatase type 1 catalytic subunit in Saccharomyces cerevisiae. AB - Mutations in GLC7, the gene encoding the type 1 protein phosphatase catalytic subunit, cause a variety of abberrant phenotypes in yeast, such as impaired glycogen synthesis and relief of glucose repression of the expression of some genes. Loss of function of the REG1/HEX2 gene, necessary for glucose repression of several genes, was found to suppress the glycogen-deficient phenotype of the glc7-1 allele. Deletion of REG1 in a wild-type background led to overaccumulation of glycogen as well as slow growth and an enlarged cell size. However, loss of REG1 did not suppress other phenotypes associated with GLC7 mutations, such as inability to sporulate or, in cells bearing the glc7Y-170 allele, lack of growth at 14 degrees. The effect of REG1 deletion on glycogen accumulation is not simply due to derepression of glucose-repressed genes, although it does require the presence of SNF1, which encodes a protein kinase essential for expression of glucose-repressed genes and for glycogen accumulation. We propose that REG1 has a role in controlling glycogen accumulation. PMID- 8722768 TI - Gene conversion alone accounts for more than 90% of recombination events at the am locus of Neurospora crassa. AB - We have used closely flanking molecular markers located approximately 4 kb distal and 6 kb proximal of the am locus to investigate the incidence of crossover events associated with the generation of prototrophic recombinants in a cross heteroallelic am1 am6. Ninety-three percent of prototrophs were generated by events that did not recombine the molecular markers, indicating that simple conversion accounts for the formation of most prototrophs and that associated crossovers are much less frequent (approximately 0.07) than estimated previously using more distant flanking markers. This suggests that conversion and crossing over during meiosis may arise from distinct mechanisms or that if, as is widely supposed, conversion and crossing over result from alternate modes of resolution of Holliday junctions then, at least for the am locus of Neurospora, the mode of resolution is strongly biased in favor of retaining the parental association of flanking sequences. Because estimates of the association of conversion and crossing over based on more distant gene markers are similar for yeast and Neurospora (approximately 0.35), our observation may have general significance. PMID- 8722769 TI - Epigenetic control of a transposon-inactivated gene in Neurospora is dependent on DNA methylation. AB - An unstable allele of the Neurospora am (GDH) gene resulting from integration of the retrotransposon Tad3-2 into 5' noncoding sequences was found in previous work. We report that reversion to Am+ depends on DNA methylation within and upstream of Tad. Levels of methylation were correlated with the proportion of Am+ conidia, whether the cultures were derived from Am- or Am+ isolates. Reversion to Am+ did not occur when conidia were plated on 5-azacytidine, which reduces DNA methylation. The mutation dim-2, which appears to abolish DNA methylation, also prevented reversion to Am+. The native am allele, in a strain that lacked Tad elements, was replaced with am::Tad3-2 or with a deletion derivative that prevents transposition of Tad. Transformants of both classes showed instability comparable with that of the original isolates, which contain multiple Tad elements. Deletion of the upstream enhancer-like sequences, URSam alpha and beta, did not prevent the instability of am::Tad3-2. The results suggest that am expression is dependent on DNA methylation but not on proliferation or transposition of the Tad element and that the instability does not require the upstream sequences of am. PMID- 8722771 TI - Suppression and enhancement of the Aspergillus nidulans medusa mutation by altered dosage of the bristle and stunted genes. AB - Asexual reproduction in Aspergillus nidulans is characterized by the orderly differentiation of multicellular reproductive structures (conidiophores) and chains of uninucleate conidia (spores). Mutations in the developmental modifier medusa (medA) result in aberrant conidiophores with branching chains of reiterated reproductive cells (metulae), delayed conidial differentiation and frequent reinitiation of secondary conidiophores. We show that incorrect morphology is in part a consequence of modified bristle (brlA) and abacus (abaA) expression, key regulators of the core genetic pathway directing conidial differentiation. First, correct temporal expression of both brlA transcripts (brlA alpha and brlA beta) requires MedAp. Second, MedAp functions as a coactivator required for normal levels of abaA expression. Finally, we show that wild-type morphology results from a finely tuned balance in the expression of brlA, medA and the developmental modifier stunted (stuA). One extra copy of brlA suppresses medA mutations and restores normal abaA mRNA abundance. In contrast, an extra copy of stuA in a medA- strain results in an enhanced medusoid phenotype with extensive metulae proliferation and nearly complete absence of conidia. abaA and brlA alpha transcription are completely repressed in these strains. In general, low stuA:brlA ratios promoted conidiation while high ratios caused proliferation of unicellular sterigmata and inhibited conidiation. PMID- 8722770 TI - The vacuolar ATPase of Neurospora crassa is indispensable: inactivation of the vma-1 gene by repeat-induced point mutation. AB - To analyze the phenotype of cells lacking the vacuolar ATPase, we inactivated the vma-1 gene, which encodes the catalytic subunit of the enzyme. Because preliminary experiments suggested the vma-1 gene was essential, we developed a method of simultaneously inactivating the gene and complementing it with a functional copy. We call this method repeat-induced point mutation (RIP) & Rescue. Two strains, both of which contained an extra copy of the vma-1 gene, were mated. Progeny that had inherited a functional copy of the gene at an ectopic site in the genome were selected. In some of these progeny the endogenous vma-1 gene had been altered by the RIP process. Sequencing showed the endogenous vma-1 gene had been inactivated by multiple point mutations. Progeny from strains with an inactive endogenous vma-1 gene were inviable unless a functional copy of the gene cosegregated, indicating that the vacuolar ATPase is essential in Neurospora crassa. PMID- 8722772 TI - The REC1 gene of Ustilago maydis, which encodes a 3'-->5' exonuclease, couples DNA repair and completion of DNA synthesis to a mitotic checkpoint. AB - Mutation in the REC1 gene of Ustilago maydis results in extreme sensitivity to killing by ultraviolet light. The lethality of the rec1-1 mutant was found to be partially suppressed if irradiated cells were held artificially in G2-phase by addition of a microtubule inhibitor. This mutant was also found to be sensitive to killing when DNA synthesis was inhibited by external means through addition of hydroxyurea or by genetic control in a temperature-sensitive mutant strain defective in DNA synthesis. Flow cytometric analysis of exponentially growing cultures indicated that wild-type cells accumulated in G2 after UV irradiation, while rec1-1 cells appeared to exit from G2 and accumulate in G1/S. Analysis of mRNA levels in synchronized cells indicated that the REC1 gene is periodically expressed with the cell cycle and reaches maximal levels at G1/S. The results are interpreted to mean that a G2-M checkpoint is disabled in the rec1-1 mutant. It is proposed that the REC1 gene product functions in a surveillance system operating during S-phase and G2 to find and repair stretches of DNA with compromised integrity and to communicate with the cell cycle apparatus. PMID- 8722773 TI - A genetic map of Gibberella fujikuroi mating population A (Fusarium moniliforme). AB - We constructed a recombination-based map of the fungal plant pathogen Gibberella fujikuroi mating population A (asexual stage Fusarium moniliforme). The map is based on the segregation of 142 restriction fragment length polymorphism (RFLP) markers, two auxotrophic genes (arg1, nic1), mating type (matA+/matA-), female sterility (ste1), spore-killer (Sk), and a gene governing the production of the mycotoxin fumonisin B1 (fum1) among 121 random ascospore progeny from a single cross. We identified 12 linkage groups corresponding to the 12 chromosome-sized DNAs previously observed in contour-clamped homogeneous electric field (CHEF) gels. Linkage groups and chromosomes were correlated via Southern blots between appropriate RFLP markers and the CHEF gels. Eleven of the 12 chromosomes are meiotically stable, but the 12th (and smallest) is subject to deletions in 3% (4/121) of the progeny. Positive chiasma interference occurred on five of the 12 chromosomes, and nine of the 12 chromosomes averaged more than one crossover per chromosome. The average kb/cM ratio in this cross is approximately 32. PMID- 8722774 TI - A Mendelian mutation affecting mating-type determination also affects developmental genomic rearrangements in Paramecium tetraurelia. AB - In Paramecium tetraurelia, mating type is determined during the differentiation of the somatic macronucleus from a zygotic nucleus genetically competent for both types, O and E. Determination of the developing macronucleus is controlled by the parental macronucleus through an unknown mechanism resulting in the maternal inheritance of mating types. The pleiotropic mutation mtFE affects macronuclear differentiation. Determination for E is constitutive in mutant homozygotes; a number of unrelated mutant characters are also acquired during development. We have examined the possibility that the mutation causes a defect in the developmental rearrangements of the germ-line genome. We show that the excision of an IES (internal eliminated sequence) interrupting the coding sequence of a surface antigen gene is impaired in the mutant, resulting in an alternative macronuclear version of the gene. Once established, the excision defect is indefinitely transmitted across sexual generations in the cytoplasmic lineage, even in a wild-type genetic context. Thus, the processes of mating-type determination and excision of this IES, in addition to their common sensitivity to the mtFE mutation, show a similar maternal inheritance of developmental alternatives in wild-type cells, suggesting a molecular model for mating-type determination. PMID- 8722775 TI - Change of genetic architecture in response to sex. AB - A traditional view is that sexual reproduction increases the potential for phenotypic evolution by expanding the range of genetic variation upon which natural selection can act. However, when nonadditive genetic effects and genetic disequilibria underlie a genetic system, genetic slippage (a change in the mean genotypic value contrary to that promoted by selection) in response to sex may occur. Additionally, depending on whether natural selection is predominantly stabilizing or disruptive, recombination may either enhance or reduce the level of expressed genetic variance. Thus, the role of sexual reproduction in the dynamics of phenotypic evolution depends heavily upon the nature of natural selection and the genetic system of the study population. In the present study, on a permanent lake Daphnia pulicaria population, sexual reproduction results in significant genetic slippage and a significant increase in expressed genetic variance for several traits. These observations provide evidence for substantial genetic disequilibria and nonadditive genetic effects underlying the genetic system of the study population. From these results, the fitness function of the previous clonal selection phase is inferred to be directional and/or stabilizing. The data are also used to infer the effects of natural selection on the mean and the genetic variance of the population. PMID- 8722776 TI - Genetic and molecular analysis of spe-27, a gene required for spermiogenesis in Caenorhabditis elegans hermaphrodites. AB - Hermaphrodites with mutations in the spe-27 gene are self-sterile, laying only unfertilized eggs; mutant males are fertile. Hermaphrodites make spermatids that fail to activate to crawling spermatozoa so passing oocytes sweep them out of the spermatheca. These spermatids do activate and produce self-progeny if young mutant hermaphrodites are mated by fertile (or sterile) males. Spermatids isolated from either mutant males or hermaphrodites initiate activation in vitro when treated with proteases, but then arrest with spiky membrane projections that resemble those of a normal intermediate in pseudopod formation. These phenotypes are identical to spe-8 and spe-12 mutants. They can be explained if males and hermaphrodites have distinct pathways for spermatid activation, and these three genes are necessary only for the hermaphrodite pathway. Consistent with this model, when spe-27 mutant male spermatids without seminal fluid are artificially inseminated into hermaphrodites, they fail to activate. The spe-27 gene has been isolated, sequenced and its regulatory regions identified. The sequence predicts a 131 amino acid polypeptide that has no striking structural motifs and no resemblance to known proteins. Two of the mutations in spe-27 alter mRNA splicing; a third mutation is a temperature-sensitive missense mutation. PMID- 8722777 TI - Suppressors of the unc-73 gene of Caenorhabditis elegans. AB - The unc-73 gene of Caenorhabditis elegans is necessary for proper axon guidance. Animals mutant in this gene are severely uncoordinated and also exhibit defects in cell migration and cell lineages. We have isolated coordinated revertants of unc-73 (e936). These fall into three classes: intragenic revertants, extragenic dominant suppressors (sup-39), and a single apparently intragenic mutation that is a dominant suppressor with a linked recessive lethal phenotype. sup-39 mutations cause early embryonic lethality, but escapers have a wild-type movement phenotype as larvae and adults. Gonads of sup-39 mutant animals show a novel defect: normal gonads have a single row of oocytes, but sup-39 gonads often have two rows of oocytes. This result suggests that the mutant gonad is defective in choosing on its surface only a single site form which nuclei will emerge to form oocytes. These results are interpreted in terms of an effect of unc-73 on determination of cell polarity. PMID- 8722778 TI - The Caenorhabditis elegans sel-1 gene, a negative regulator of lin-12 and glp-1, encodes a predicted extracellular protein. AB - The Caenorhabditis elegans lin-12 and glp-1 genes encode members of the LIN 12/NOTCH family of receptors. The sel-1 gene was identified as an extragenic suppressor of a lin-12 hypomorphic mutant. We show in this report that the sel-1 null phenotype is wild type, except for an apparent elevation in lin-12 and glp-1 activity in sensitized genetic backgrounds, and that this genetic interaction seems to be lin-12 and glp-1 specific. We also find that sel-1 encodes a predicted extracellular protein, with a domain sharing sequence similarity to predicted proteins from humans and yeast. SEL-1 may interact with the LIN-12 and GLP-1 receptors and/or their respective ligands to down-regulate signaling. PMID- 8722779 TI - Single amino acid mutations in Drosophila fascin disrupt actin bundling function in vivo. AB - Fascins bundle actin filaments into large, tightly packed hexagonal arrays that support diverse cellular processes including microvillar projections and filopodial extensions. In Drosophila, fascin is encoded by the singed locus. Severe singed mutants have gnarled bristles and are female sterile due to a defect in rapid cytoplasm transport during oogenesis. In this paper, we report the results of a large EMS mutagenesis screen to generate new singed alleles. A mutation that changes glycine 409 to glutamic acid results in partial inactivation of fascin in vivo; singedG409E mutants have kinked bristles and are fertile with a mild nurse cell cytoplasm transport defect. This mutation is in a small conserved domain near the C-terminus of fascin. A mutation that changes serine 289 to asparagine almost completely inactivates fascin in vivo; singedS289N mutants have gnarled bristles and are sterile due to a severe defect in nurse cell cytoplasm transport caused by the absence of nurse cell cytoplasmic actin bundles. A subsequent EMS mutagenesis screen for dominant suppressors of singedS289N sterility revealed an intragenic suppressor mutation that changes serine 251 to phenylalanine and restores much of fascin's function. These two mutations, S289N and S251F, draw attention to a central domain in fascin. PMID- 8722780 TI - Bioassaying putative RNA-binding motifs in a protein encoded by a gene that influences courtship and visually mediated behavior in Drosophila: in vitro mutagenesis of nonA. AB - The no-on-transient-A (nonA) gene of Drosophila melanogaster influences vision, courtship song, and viability. The nonA-encoded polypeptide is inferred to bind single-stranded nucleic acids. Although sequence-analysis of NONA implies that it belongs to a special interspecific family of this protein type, it does contain two classical RNA recognition motifs (RRM). Their behavioral significance was assayed by generating transgenic strains that were singly or multiply mutated within the relatively N-terminal motif (RRM1) or within RRM2. Neither class of mutation affected NONA binding to polytene chromosomes. The former mutations led to extremely low viability, accompanied by diminished adult longevities that were much worse than for a nonA-null mutant, implying that faulty interpolypeptide interactions might accompany the effects of the amino-acid substitutions within RRM1. All in vitro-mutated types caused optomotor blindness and an absence of transient spikes in the electroretinogram. Courtship analysis discriminated between the effects of the mutations: the RRM2-mutated type generated song pulses and trains that tended to be mildly mutant. These phenotypic abnormalities reinforce the notion that nonA's ubiquitous expression has its most important consequences in the optic lobes, the thoracic ganglia, or both, depending in part on the nonA allele. PMID- 8722781 TI - Effects of single P-element insertions on bristle number and viability in Drosophila melanogaster. AB - Single P-element mutagenesis was used to construct 1094 lines with P[lArB] inserts on all three major chromosomes in an isogenic background previously free of P elements. The effects of insertions on bristle number and on viability were assessed by comparison to 392 control lines. The variance and effects of P element inserts on bristle number and viability were larger than those inferred from spontaneous mutations. The distributions of effects on bristle number were symmetrical and highly leptokurtic, such that a few inserts with large effects caused most of the increase in variance. The distribution of effects on viability were negatively skewed and platykurtic. On average, the effects of P-element insertions on bristle number were partly recessive and on viability were completely recessive. P-element inserts with large effects on bristle number tended to have reduced viability, but the correlation between the absolute value of the effects on bristle number and on viability was not strong. Fifty P-element inserts tagging quantitative trait loci (QTLs) with large effects on bristle number were mapped cytogenetically. Two P-element-induced scabrous alleles and five extramacrochaetae alleles were generated. Single P-element mutagenesis is a powerful method for identifying QTLs at the level of genetic locus. PMID- 8722782 TI - Effects of single P-element insertions on olfactory behavior in Drosophila melanogaster. AB - Single P-element (P[lArB]) insertional mutagenesis of an isogenic strain was used to identify autosomal loci affecting odor-guided behavior of Drosophila melanogaster. The avoidance response to benzaldehyde of 379 homozygous P[lArB] element-containing insert lines was evaluated quantitatively. Fourteen smell impaired (smi) lines were identified in which P[lArB] element insertion caused different degrees of hyposmia in one or both sexes. The smi loci map to different cytological locations and probably are novel olfactory genes. Enhancer trap analysis of the smi lines indicates that expression of at least 10 smi genes is controlled by olfactory tissue-specific promoter/enhancer elements. PMID- 8722783 TI - A negative feedback mechanism revealed by functional analysis of the alternative isoforms of the Drosophila splicing regulator transformer-2. AB - The Drosophila sex determination gene transformer-2 (tra-2) is a splicing regulator that affects the sex-specific processing of several distinct pre-mRNAs. While the tra-2 gene itself is known to produce alternative mRNAs that together encode three different TRA-2 protein isoforms, the respective roles of these isoforms in affecting individual pre-mRNA targets has remained unclear. We have generated transgenic fly strains with mutations affecting specific TRA-2 isoforms to investigate their individual roles in regulating the alternative processing of doublesex, exuperantia and tra-2 pre-mRNA. Our results indicate that in somatic tissues two different isoforms function redundantly to direct female differentiation and female-specific doublesex pre-mRNA splicing. In the male germline, where tra-2 has an essential role in spermatogenesis, a single isoform was found to uniquely perform all necessary functions. This isoform appears to regulate its own synthesis during spermatogenesis through a negative feedback mechanism involving intron retention. PMID- 8722784 TI - A screen for genes that function downstream of Ras1 during Drosophila eye development. AB - Cell-fate specification of the R7 photoreceptor cell is controlled by the sevenless receptor tyrosine kinase (SevRTK) and Ras1, the Drosophila homologue of mammalian H-ras, K-ras and N-ras oncogenes. An activated form of Ras1 expressed under control of the sevenless enhancer/promoter (sev-Ras1V12) induces production of supernumerary R7 photoreceptor cells, which causes the eye to become rough in appearance. To isolate mutations in genes functioning downstream of Ras1, we carried out a screen for dominant suppressors and enhancers of this rough eye phenotype. Approximately 850,000 mutagenized flies were screened, and 282 dominant suppressors and 577 dominant enhancers were isolated. Mutations in the Drosophila homologues of Raf, MEK, MAPK, type I Geranylgeranyl Transferase and Protein Phosphatase 2A were isolated, as were mutations in several novel signaling genes. Some of these mutant genes appear to be general signaling factors that function in other Ras1 pathways, while one seems to be more specific for photoreceptor development. At least two suppressors appear to function either between Ras1 and Raf or in parallel to Raf. PMID- 8722785 TI - Courtship anomalies caused by doublesex mutations in Drosophila melanogaster. AB - The role played by the sex-determining gene doublesex (dsx) and its influence on Drosophila courtship were examined. Against a background of subnormal male-like behavior that is reported to be an attribute of haplo-X flies homozygous for the original dsx mutation, and given that a sex-specific muscle is unaffected by genetic variation at this locus, analyses of several reproductive behaviors and control for genetic background effects indicated that XY dsx mutants are impaired in their willingness to court females. When they did court, certain behavioral actions were normal, including components of courtship song. However, these mutants never produced courtship humming sounds. Mature XY dsx flies elicited anomalously high levels of courtship; that this occurs merely because of a delay in imaginal development was experimentally discounted. The current analysis reconciled two ostensibly conflicting reports involving the courtship-stimulating qualities of this mutant type. Such experiments also uncovered a new behavioral anomaly: dsx mutations caused chromosomal males to court other males at abnormally high levels. These results are discussed from the perspective of doublesex's influence on internal tissues of adult Drosophila involved in the triggering and neural control of male- and female-like elements of courtship, reproductive pheromone production, or a combination of such factors. PMID- 8722786 TI - Elimination of introns at the Drosophila suppressor-of-forked locus by P-element mediated gene conversion shows that an RNA lacking a stop codon is dispensable. AB - The suppressor of forked [su(f)] locus affects the phenotype of mutations caused by transposable element insertions at unlinked loci. It encodes a putative 84-kD protein with homology to two proteins involved in mRNA 3' end processing; the product of the yeast RNA14 gene and the 77-kD subunit of human cleavage stimulation factor. Three su(f) mRNAs are produced by alternative polyadenylation. The 2.6- and 2.9-kb mRNAs encode the same 84-kD protein while a 1.3-kb RNA, which terminates within the fourth intron, is unusual in having no stop codon. Using P-element-mediated gene replacement we have copied sequences from a transformation construct into the su(f) gene creating a su(f) allele at the normal genomic location that lacks the first five introns. This allele is viable and appears wild type for su(f) function, demonstrating that the 1.3-kb RNA and the sequences contained within the deleted introns are dispensable for su(f) function. Compared with studies on gene replacement at the white locus, chromosomal breaks at su(f) appear to be less efficiently repaired from ectopic sites, perhaps because of the location of su(f) at the euchromatin/heterochromatin boundary on the X chromosome. PMID- 8722787 TI - Genetics of differences in pheromonal hydrocarbons between Drosophila melanogaster and D. simulans. AB - Females of Drosophila melanogaster and its sibling species D. simulans have very different cuticular hydrocarbons, with the former bearing predominantly 7,11 heptacosadiene and the latter 7-tricosene. This difference contributes to reproductive isolation between the species. Genetic analysis shows that this difference maps to only the third chromosome, with the other three chromosomes having no apparent effect. The D. simulans alleles on the left arm of chromosome 3 are largely recessive, allowing us to search for the relevant regions using D. melanogaster deficiencies. At least four nonoverlapping regions of this arm have large effects on the hydrocarbon profile, implying that several genes on this arm are responsible for the species difference. Because the right arm of chromosome 3 also affects the hydrocarbon profile, a minimum of five genes appear to be involved. The large effect of the third chromosome on hydrocarbons has also been reported in the hybridization between D. simulans and its closer relative D. sechellia, implying either an evolutionary convergence or the retention in D. sechellia of an ancestral sexual dimorphism. PMID- 8722788 TI - Germline transformation of Drosophila virilis with the transposable element mariner. AB - An important goal in molecular genetics has been to identify a transposable element that might serve as an efficient transformation vector in diverse species of insects. The transposable element mariner occurs naturally in a wide variety of insects. Although virtually all mariner elements are nonfunctional, the Mos1 element isolated from Drosophila mauritiana is functional. Mos1 was injected into the pole-cell region of embryos of D. virilis, which last shared a common ancestor with D. mauritiana 40 million years ago. Mos1 PCR fragments were detected in several pools of DNA from progeny of injected animals, and backcross lines were established. Because G0 lines were pooled, possibly only one transformation event was actually obtained, yielding a minimum frequency of 4%. Mos1 segregated in a Mendelian fashion, demonstrating chromosomal integration. The copy number increased by spontaneous mobilization. In situ hybridization confirmed multiple polymorphic locations of Mos1. Integration results in a characteristic 2-bp TA duplication. One Mos1 element integrated into a tandem array of 370-bp repeats. Some copies may have integrated into heterochromatin, as evidenced by their ability to support PCR amplification despite absence of a signal in Southern and in situ hybridization. PMID- 8722789 TI - Nucleotide variation at the Gpdh locus in the genus Drosophila. AB - The Gpdh locus was sequenced in a broad range of Drosophila species. In contrast to the extreme evolutionary constraint seen at the amino acid level, the synonymous sites evolve at rates comparable to those of other genes. Gpdh nucleotide sequences were used to infer a phylogenetic tree, and the relationships among the species of the obscura group were examined in detail. A survey of nucleotide polymorphism within D. pseudoobscura revealed no amino acid variation in this species. Applying a modified McDonald-Kreitman test, the amino acid divergence between species in the obscura group does not appear to be excessive, implying that drift is adequate to explain the patterns of amino acid change at this locus. In addition, the level of polymorphism at the Gpdh locus in D. pseudoobscura is comparable to that found at other loci, as determined by a Hudson-Kreitman-Aguade test. Thus, the pattern of nucleotide variation within and between species at the Gpdh locus is consistent with a neutral model. PMID- 8722790 TI - A quantitative measure of the mitotic pairing of alleles in Drosophila melanogaster and the influence of structural heterozygosity. AB - In Drosophila there exist several examples of gene expression that can be modified by an interaction between alleles; this effect is known as transvection. The inference that alleles interact comes from the observations that homologous chromosomes pair in mitotically dividing cells, and that chromosome rearrangements can alter the phenotype produced by a pair of alleles. It is thought that heterozygous rearrangements impede the ability of alleles to pair and interact. However, because the existing data are inconsistent, this issue is not fully settled. By measuring the frequency of site-specific recombination between homologous chromosomes, we show that structural heterozygosity inhibits the pairing of alleles that lie distal to a rearrangement breakpoint. We suggest that some of the apparent conflicts may owe to variations in cell-cycle lengths in the tissues where the relevant allelic interactions occur. Cells with a longer cell cycle have more time to establish the normal pairing relationships that have been disturbed by rearrangements. In support, we show that Minute mutations, which slow the rate of cell division, partially restore a transvection effect that is disrupted by inversion heterozygosity. PMID- 8722791 TI - Amino acid polymorphism and rare electrophoretic variants of G6PD from natural populations of Drosophila melanogaster. AB - Identifying the amino acid changes responsible for electrophoretic variants is essential to understanding the significance of allozyme polymorphism in adaptation. The amino acid mutations responsible for the common G6PD allozyme polymorphisms in Drosophila melanogaster have been recently described. This study characterizes the amino acid changes associated with 11 rare electrophoretic G6PD variants. The 11 rare electrophoretic variants result from six independent amino acid mutations. The in vivo function of the rare variants was determined in an earlier study and most variants fell into one of two function classes. It is shown here that the function of the rare variants reflects the state of the Pro/Leu mutation responsible for the A/B allozyme polymorphism in each variant. Two mutations destabilize quaternary structure resulting in shifts from tetrameric dimeric alleles, and one of these also results in a variant with in vivo function intermediate to A and B. That mutation is an aspartic-acid-to asparagine change that is two residues away from the Pro/Leu polymorphism responsible for the A/B dimertetramer quaternary shift. Structure-function relationships based on studies of human G6PD deficiency-associated mutations predict that these last two amino acid changes fall within the protein domain responsible for NADP binding. PMID- 8722792 TI - Dominance of insecticide resistance presents a plastic response. AB - Dominance level of insecticide resistance provided by one major gene (an insensitive acetylcholinesterase) in the mosquito Culex pipiens was studied in two distinct environments. Dominance level was found to be very different environments, varying from almost complete dominance to almost recessive when either propoxur (a carbamate insecticide) or chlorpyrifos (an organophosphorus insecticide) was used. To better understand this plastic response, three environmental parameters were manipulated and their interactions studied. For chlorpyrifos, each parameter had a small effect, but when all parameters were changed, the dominance level was greatly affected. For propoxur, one environmental parameter had a large effect by itself. It was further studied to understand the causal relationship of this plasticity. Recessivity of resistance was associated with more demanding environments. These results are discussed in the context of the various theories of the evolution of dominance. It appears that dominance of insecticide resistance cannot be directly predicted by Wright's physiological theory. PMID- 8722793 TI - Woot, an active gypsy-class retrotransposon in the flour beetle, Tribolium castaneum, is associated with a recent mutation. AB - A recently isolated, lethal mutation of the homeotic Abdominal gene of the red flour beetle Tribolium castaneum is associated with an insertion of a novel retrotransposen into an intron. Sequence analysis indicates that this retrotransposon, named Woot, is a member of the gypsy family of mobile elements. Most strains of T. castaneum appear to harbor approximately 25-35 copies of Woot per genome. Woot is composed of long terminal repeats of unprecedented length (3.6 kb each), flanking an internal coding region 5.0 kb in length. For most copies of Woot, the internal region includes two open reading frames (ORFs) that correspond to the gag and pol genes of previously described retrotransposons and retroviruses. The copy of Woot inserted into Abdominal bears an apparent single frameshift mutation that separates the normal second ORF into two. Woot does not appear to generate infectious virions by the criterion that no envelop gene is discernible. The association of Woot with a recent mutation suggests that this retroelement is currently transpositionally active in at least some strains. PMID- 8722794 TI - Mitochondrial DNA sequence variation in the eastern house mouse, Mus musculus: comparison with other house mice and report of a 75-bp tandem repeat. AB - The control region and flanking tRNAs were sequenced from 139 Mus musculus mitochondrial DNAs (mtDNAs) from mice collected at 44 localities extending from Germany to Japan. Among the 36 types of M. musculus mtDNA resolved, five have an added 75-bp direct repeat; the two copies within an individual differ by two to four base substitutions. Among 90 M. domesticus mtDNAs sequenced, 12 new types were found; 96 M. domesticus types have now been identified by sequencing this segment. Representative mtDNAs from M. castaneus, M. macedonicus, M. spicilegus and M. spretus were also sequenced. A parsimony tree for the M. musculus mtDNAs is about half as deep as the tree for the M. domesticus mtDNAs, which is consistent with the idea that M. musculus is genetically less diverse and younger than M. domesticus. The patterns of variation as a function of position are similar but not identical in M. musculus and M. domesticus mtDNAs. M. castaneus and M. musculus mtDNAs are allied, at a tree depth about three times as great as the start of intra-M. musculus divergence. The coalescence of the M. musculus and M. castaneus mtDNAs is about half as deep as their coalescence with the M. domesticus mtDNA lineages. The mtDNAs of the aboriginal M. macedonicus and M. spicilegus are each other's closest relatives, at a tree depth greater than the deepest intracommensal node. The mtDNA results support the view that the aboriginal M. spretus is the sister group of the other five species. PMID- 8722795 TI - Complementation mapping of skeletal and central nervous system abnormalities in mice of the piebald deletion complex. AB - The s15DttMb, s36Pub, s1Acrg and s24Pub piebald deletion alleles belong to a set of overlapping deficiencies on the distal portion of chromosome 14. Molecular analysis was used to define the extent of the deletions. Mice homozygous for the smallest deletion, s15DttMb, die shortly after delivery and display alterations in the central nervous system, including hydrocephalus and a dorsally restricted malformation of the spinal cord. These mice also display homeotic transformations of vertebrae in the midthoracic and lumbar regions. Homozygous s27Pub mice contain a point mutation in the piebald gene, survive to weaning, and display no central nervous system or skeletal defects, arguing that the s15DttMb phenotype results from the loss of genes in addition to piebald. A larger deletion, s36Pub, exhibits additional cartilage malformations and defects in the anterior axial and cranial skeleton. The skeletal defects in both s15DttMb and s36Pub mice resemble transformations associated with the targeted disruption of Hox genes and genes encoding the retinoic acid receptors, which play a role in the specification of segmental identity along the anteroposterior axis. Complementation analysis of the s15DttMb and s36Pub phenotypes, using two additional deletions, localized the gene(s) associated with each phenotype to a defined chromosomal region. PMID- 8722796 TI - Genotype-environment interaction: apolipoprotein E (ApoE) gene effects and age as an index of time and spatial context in the human. AB - We analyzed the age-dependence of the estimates of the parameters of the genetic architecture of plasma ApoE levels associated with ApoE gene variation. Our study sample included 1988 individuals in multigeneration pedigrees from the Rochester, MN, population. We used a 30-yr sliding window across the age range (5-90 yr) to estimate the age dependency of parameters. Additive ApoE allelic variance of transformed plasma ApoE values for both genders, heritabilities for males and phenotypic and residual variance for females peaked in the 20-40-yr age windows and decreased significantly with age (P < 0.05). Phenotypic and residual variance for males and dominance variance for both genders did not vary significantly with age. All parameter estimates were significantly different from zero across all age windows for both genders. Most studies of ApoE have focused on its functions in the pathophysiology of coronary artery disease (CAD) in middle-aged and older individuals. Our findings suggest the greatest role of this gene is in determining phenotype differences among younger and middle-aged individuals. These observed genotypic effects on the plasma ApoE levels may contribute to age dependent differences in physiological health, growth, and risk of disease. PMID- 8722798 TI - Mosaic analysis of the liguleless3 mutant phenotype in maize by coordinate suppression of mutator-insertion alleles. AB - Liguleless3-O (Lg3-O) transforms the leaf blade, auricle and ligule into sheath around the midrib region. We conducted a genetic mosaic analysis of the Lg3 phenotype to determine the site of Lg3 gene action. Combining the Mutator (Mu) suppressible Lg3-Or211 and a1-mum2 alleles in a Mu-active background generated a stock wherein somatic loss of Mu activity resulted in anthocyanin-marked clonal sectors expressing Lg3 in the leaf. Lg3-Or211 plants appear wild type in a Mu active line, but Mu-inactive plants express a severe Lg3 phenotype. We observed four sector classes: wild type, sheath-like with ligule displacement, sheath-like with ectopic ligule, and auricle-like. The mutation does not cause transformation to a specific cell or regional identity. Lg3-Or211 activity in the mesophyll alters wild-type epidermal cell fates; activity in epidermis seems functionless. Lg3 mutant activity has a nonautonomous, cell-layer-specific function in the transverse dimension. In the lateral dimension, sectors of Lg3 mutant phenotype can exhibit either cell-autonomous or nonautonomous effects. Our work demonstrates that mosaic analysis by coordinate suppression of Mu-induced alleles is useful for analyzing the cell autonomy of genetically defined functions. PMID- 8722797 TI - The y1 gene of maize codes for phytoene synthase. AB - The cloned y1 locus of maize was sequenced and found to encode phytoene synthase. Different "wild-type" alleles of the locus were found to differ by the insertion of transposable elements in their promoter and polyA addition regions, and by the length of a CCA tandem repeat series, without any obvious effect on function of the gene. A dominant Y1 ("wild-type") allele was observed to be expressed at highest levels in the seedling but also in the embryo and endosperm. The Mu3 transposable element insertion responsible for a pastel allele of y1, which gives lowered levels of carotenoids in the endosperm of kernels and seedlings grown at high temperatures, was located in the 5' end of the gene. Although the size of the transcript from this y1 mutation suggests that the Mu3 element provides the promoter for this allele, leaf tissue in this mutant line contained approximately normal amounts of y1 mRNA. A recessive allele of y1, which conditions normal levels of carotenoids in the embryo and seedling, but almost no carotenoids in the endosperm, was found to accumulate normal amounts of y1 mRNA in the seedling and embryo, while y1 transcripts were not detected in the endosperm. PMID- 8722799 TI - Nuclear genes associated with a single Brassica CMS restorer locus influence transcripts of three different mitochondrial gene regions. AB - Previous studies have shown that the mitochondrial orf224/atp6 gene region is correlated with the Polima (pol) cytoplasmic male sterility (CMS) of Brassica napus. We now extend this correlation by showing that the effects of nuclear fertility restoration on orf224/atp6 transcripts cosegregate with the pol restorer gene Rfp1 in genetic crosses. We also show, however, that the recessive rfp1 allele, or a very tightly linked gene, acts as a dominant gene, designated Mmt (modifier of mitochondrial transcripts), in controlling the presence of additional smaller transcripts of the nad4 gene and a gene possibly involved in cytochrome c biogenesis. A common sequence, TTGTGG, maps immediately downstream of the 5' termini of both of the transcripts specific to plants with the Mmt gene and may serve as a recognition motif in generation of these transcripts. A similar sequence, TTGTTG, that may be recognized by the product of the alternate allele (or haplotype), Rfp1, is found within orf224 just downstream of the major 5' transcript terminus specific to fertility restored plants. Our results suggest that Rfp1/ Mmt is a novel nuclear genetic locus that affects the expression of multiple mitochondrial gene regions, with different alleles or haplotypes exerting specific effects on different mitochondrial genes. PMID- 8722800 TI - Secondary trisomics and telotrisomics of rice: origin, characterization, and use in determining the orientation of chromosome map. AB - Secondary trisomics and telotrisomics representing the 12 chromosomes of rice were isolated from the progenies of primary trisomics. A large population of each primary trisomic was grown. Plants showing variation in gross morphology compared to the primary trisomics and disomic sibs were selected and analyzed cytologically at diakinesis and pachytene. Secondary trisomics for both arms of chromosomes 1, 2, 6, 7 and 11 and for one arm of chromosomes 4, 5, 8, 9 and 12 were identified. Telotrisomics for short arm of chromosomes 1, 8, 9 and 10 and for long arms of chromosomes 2, 3 and 5 were isolated. These secondary and telotrisomics were characterized morphologically and for breeding behavior. Secondary trisomics 2n + 1S.1S, 2n + 1L.1L, 2n + 2S.2S, 2n + 2L.2L, 2n + 6S.6S, 2n + 6L.6L and 2n + 7L.7L are highly sterile, and 2n + 1L.1L, 2n + 2L.2L and 2n + 7L.7L do not set any seed even upon backcrossing. Telotrisomics are fertile and vigorous. Genetic segregation of 43 marker genes was studied in the F2 or backcross progenies. On the basis of segregation data, these genes were delimited to specific chromosome arms. Correct orientation of 10 linkage groups was determined and centromere positions on nine linkage groups were approximated. A revised linkage map of rice is presented. PMID- 8722801 TI - Molecular genetics of rust resistance in poplars (Melampsora larici-populina Kleb/Populus sp.) by bulked segregant analysis in a 2 x 2 factorial mating design. AB - With random amplified polymorphic DNA (RAPD) markers, we have tagged a genomic region in Populus sp. involved in qualitative resistance to Melampsora larici populina. Our approach was based on three steps: use of RAPD markers that can be quickly and efficiently researched: application of "bulked segregant analysis" technique on individuals of one interspecific family P. trichocarpa x P. deltoides to search for RAPD markers linked to resistance; and validation of these markers in two other families linked with the first one in a 2 x 2 factorial mating design. Of five detected markers, only one marker M03/04_480 was polymorphic in the three segregating families, involving 89 individuals and four different parents. We have estimated the recombination value of 1 cM with 1 cM sampling error. PMID- 8722802 TI - Patterns of nucleotide substitution in mitochondrial protein coding genes of vertebrates. AB - Maximum likelihood methods were used to study the differences in substitution rates among the four nucleotides and among different nucleotide sites in mitochondrial protein-coding genes of vertebrates. In the 1st + 2nd codon position data, the frequency of nucleotide G is negatively correlated with evolutionary rates of genes, substitution rates vary substantially among sites, and the transition/transversion rate bias (R) is two to five times larger than that expected at random. Generally, largest transition biases and greatest differences in substitution rates among sites are found in the highly conserved genes. The 3rd positions in placental mammal genes exhibit strong nucleotide composition biases and the transitional rates exceed transversional rates by one to two orders of magnitude. Tamura-Nei and Hasegawa-Kishino-Yano models with gamma distributed variable rates among sites (gamma parameter, alpha) adequately describe the nucleotide substitution process in 1st+2nd position data. In these data, ignoring differences in substitution rates among sites leads to largest biases while estimating substitution rates. Kimura's two-parameter model with variable-rates among sites performs satisfactorily in likelihood estimation of R, alpha, and overall amount of evolution for 1st+2nd position data. It can also be used to estimate pairwise distances with appropriate values of alpha for a majority of genes. PMID- 8722803 TI - Dynamics of repeat polymorphisms under a forward-backward mutation model: within- and between-population variability at microsatellite loci. AB - Suggested molecular mechanisms for the generation of new tandem repeats of simple sequences indicate that the microsatellite loci evolve via some of forward backward mutation. We provide a mathematical basis for suggesting a measure of genetic distance between populations based on microsatellite variation. Our results indicate that such a genetic distance measure can remain proportional to the divergence time of populations even when the forward-backward mutations produce variable and/or directionally biased alleles size changes. If the population size and the rate of mutation remain constant, then the measure will be proportional to the time of divergence of populations. This genetic distance is expressed in terms of a ratio of components of variance of allele sizes, based on expressions developed for studying population dynamics of quantitative traits. Application of this measure to data on 18 microsatellite loci in the nine human populations leads to evolutionary trees consistent with the known ethnohistory of the populations. PMID- 8722804 TI - New statistical tests of neutrality for DNA samples from a population. AB - The purpose of this paper is to develop statistical tests of the neutral model of evolution against a class of alternative models with the common characteristic of having an excess of mutations that occurred a long time ago or a reduction of recent mutations compared to the neutral model. This class of population genetics models include models for structured populations, models with decreasing effective population size and models of selection and mutation balance. Four statistical tests were proposed in this paper for DNA samples from a population. Two of these tests, one new and another a modification of an existing test, are based on EWENS' sampling formula, and the other two new tests make use of the frequencies of mutations of various classes. Using simulated samples and regression analyses, the critical values of these tests can be computed from regression equations. This approach for computing the critical values of a test was found to be appropriate and quite effective. We examined the powers of these four tests using simulated samples from structured populations, populations with linearly decreasing sizes and models of selection and mutation balance and found that they are more powerful than existing statistical tests of the neutral model of evolution. PMID- 8722805 TI - A generalization of the mixture model in the mapping of quantitative trait loci for progeny from a biparental cross of inbred lines. AB - The recent advent of molecular markers has created a great potential for the understanding of quantitative inheritance. In parallel to rapid developments and improvements in molecular marker technologies, biometrical models have been constructed, refined and generalized for the mapping of quantitative trait loci (QTL). However, current models present restrictions in terms of breeding designs to which they apply. In this paper, we develop an approach for the generalization of the mixture model for progeny from a single bi-parental cross of inbred lines. Detailed derivations are given for genetic designs involving populations developed by selfing, i.e., where marker genotypes are obtained from Fx (x > or = 2) individuals and where phenotypes are measured on Fy (y > or = x) individuals or families. Extensions to designs involving doubled-haploids, backcross-derived individuals and random matings are outlined. The derivations presented here can easily be combined with current QTL mapping approaches. PMID- 8722806 TI - Gene genealogies within mutant allelic classes. AB - A coalescent theory of the gene genealogy within an allelic class that arises by a unique mutational event is developed and analyzed. To interpret this theory it was necessary to expand on existing theory for populations of varying size. Two features of the gene genealogy--the average pairwise distance and the total tree length--within the mutant class and within the nonmutant class are found. An index, I, is proposed that describes the extent to which a genealogy is similar to one from a population of constant size (for which I = 0) or to a star genealogy (for which I = 1). The value of I is positive in growing populations and is generally positive for the gene genealogy for the mutant class. The value of I is negative for a population decreasing in size and for the nonmutant class, if the mutant arose recently. The results are discussed in the context of the infinite sites model of mutation, which is appropriate for nucleotide sequence data, and the generalized stepwise mutation model, which is appropriate for microsatellite loci. The same genealogical methods are used to find the probability of at least one recombination event between the nucleotide that defines an allelic class and a marker at a nearby linked site. PMID- 8722807 TI - Detecting heterogeneity of substitution along DNA and protein sequences. AB - Regions of differing constraint, mutation rate or combination along a sequence of DNA or amino acids lead to a nonuniform distribution of polymorphism within species or fixed differences between species. The power of five tests to reject the null hypothesis of a uniform distribution is studied for four classes of alternate hypothesis. The tests explored are the variance of interval lengths; a modified variance test, which includes covariance between neighboring intervals; the length of the longest interval; the length of the shortest third-order interval; and a composite test. Although there is no uniformly most powerful test over the range of alternate hypotheses tested, the variance and modified variance tests usually have the highest power. Therefore, we recommend that one of these two tests be used to test departure from uniformity in all circumstances. Tables of critical values for the variance and modified variance tests are given. The critical values depend both on the number of events and the number of positions in the sequence. A computer program is available on request that calculates both the critical values for a specified number of events and number of positions as well as the significance level of a given data set. PMID- 8722808 TI - Resurrecting Muller's theory of Haldane's rule. PMID- 8722809 TI - Maternal effect as a cause of exceptions for Haldane's rule. PMID- 8722810 TI - Dominance and Haldane's rule. PMID- 8722811 TI - Apparent stabilizing selection and the maintenance of neutral genetic variation. PMID- 8722812 TI - Short echo time proton magnetic resonance spectroscopic imaging of macromolecule and metabolite signal intensities in the human brain. AB - A novel approach is presented for imaging macromolecule and metabolite signals in brain by proton magnetic resonance spectroscopic imaging. The method differentiates between metabolites and macromolecules by T1 weighting using an inversion pulse followed by a variable inversion recovery time before localization and spectroscopic imaging. In healthy subjects, the major macromolecule resonances at 2.05 and 0.9 ppm were mapped at a nominal spatial resolution of 1 x 1 x 1.5 cm3 and were demonstrated to be highly reproducible between subjects. In subacute stroke patients, a highly elevated macromolecule resonance at 1.3 ppm was mapped to infarcted brain regions, suggesting potential applications for studying pathological conditions. PMID- 8722813 TI - Simultaneous multicompartment intracellular Ca2+ measurements in the perfused heart using 19F NMR spectroscopy. AB - Although Ca2+ transport regulation at subcellular organelles is of great interest, only limited methodology has been available for measuring organellar [Ca2+] levels. The present study employs the 19F NMR resonance frequency of 4F BAPTA to measure free [Ca2+]. In 4F-BAPTA loaded perfused rabbit hearts, two 19F NMR resonances were clearly observed. The frequency of one was consistent with cytosolic [Ca2+] levels. Responses to agents that after sarcoplasmic reticulum function identified the other resonance as originating from that organelle. The experiments demonstrate the utility of NMR shift indicator methodology in obtaining simultaneous multi-compartment intracellular [Ca2+] measurements and in enabling organellar [Ca2+] measurements to be made from within intact living tissue. PMID- 8722814 TI - A new ytterbium chelate as contrast agent in chemical shift imaging and temperature sensitive probe for MR spectroscopy. AB - A paramagnetic Yb(III) complex that is the prototype of a novel class of probes for MRI and MRS has been developed. The complex displays highly shifted 1H resonances that are characterized by short relaxation times and, as such, may prove to be a valuable alternative in applications that currently require fluorine-containing probes. Selective excitation of the paramagnetically shifted resonances allows the spatial distribution of the complex to be mapped. This communication reports the images that were obtained by selectively exciting the most intense methyl group (-14.2 ppm at 27 degrees C) for complex concentrations ranging from 0.003-0.1 M. Spectroscopically, the complex may be used as a temperature probe since the proton chemical shifts exhibit a strong temperature dependence. In human serum the chemical shift difference of a selected pair of proton resonances was observed to follow a gradient of -0.42 +/- 0.01 ppm/degrees C. Furthermore, since the chemical shift of the methyl resonance displays a temperature coefficient of -0.04 +/- 0.01 ppm/degrees C, it should be possible to use the image phase for thermal mapping. PMID- 8722815 TI - Single-shot, B1-insensitive slice selection with a gradient-modulated adiabatic pulse, BISS-8. AB - An adiabatic pulse has been developed to accomplish uniform slice-selective excitation with a spatially inhomogeneous B1. This new pulse can generate a uniform, arbitrary flip angle that is determined by four adjustable phase shifts in the pulse. Self-refocused slice selection is achieved by modulating a B(O) gradient in concert with the pulse frequency (or phase) modulation. B1 compensated, self-refocused slice selection is demonstrated in computer simulations and phantom experiments using a surface transmitter/receiver coil. This adiabatic pulse can provide optimal performance in multislice MRI and localized spectroscopy when transmitting with an inhomogeneous B1. PMID- 8722816 TI - Abnormal cerebral metabolism in polydrug abusers during early withdrawal: a 31P MR spectroscopy study. AB - Phosphorus magnetic resonance spectroscopy (31P MRS) at 1.5 T was performed on nine polysubstance abusing men. All nine patients met DSM-III-R criteria for concurrent cocaine and heroin dependence, were neurologically normal, were negative for the human immunodeficiency virus, and had normal clinical brain MRI scans. Patients were scanned 2-7 days after admission to a drug treatment unit. Eleven age-matched control subjects also were studied. The ISIS localized phosphorus spectra were obtained from a 5-cm thick axial brain slice and a 100-cc white matter volume. In the brain slice, the phosphorus metabolite signal expressed as a percentage of total phosphorus signal was 15% higher for phosphomonoesters, 10% lower for nucleotide triphosphates (beta-NTP), and 7% lower for total nucleotide phosphates in polydrug abusers compared with those in controls. Phosphodiesters, inorganic phosphate, phosphocreatine, total phosphorus, pH, and free magnesium concentration were unchanged. None of these parameters correlated with the methadone dose or the number of days abstinence. Single photon emission computed tomographic imaging of a subgroup of the patients revealed abnormal cerebral perfusion in 80% of the patients scanned. These data suggest that cerebral high energy phosphate and phospholipid metabolite changes result from long term drug abuse and/or withdrawal and that these changes can be detected and studied by 31P MRS. PMID- 8722817 TI - Human cardiac high-energy phosphate metabolite concentrations by 1D-resolved NMR spectroscopy. AB - We have developed a method that can measure high-energy phosphate metabolite concentrations in humans with 1D resolved surface-coil NMR spectroscopy. The metabolites are measured by phosphorus (31P) NMR spectroscopy, and the tissue water proton (1H) resonance from the same volume serves as an internal concentration reference. The method requires only the additional acquisition of a 1H data set, and a simple calibration, performed separately, to determine the ratio of the signal per proton to the signal per phosphorus nucleus. The quantification method is particularly useful for human cardiac spectroscopy, where it eliminates image-based tissue volumetry and the corrections for signal sensitivity and phase nonuniformity necessary in prior approaches. Corrections are introduced to account for blood and fat contributions to the spectra. The method was validated on phantoms of phosphate of varying concentrations and on the human calf muscle. In calf, the adenosine triphosphate (ATP) and phosphocreatine (PCr) concentrations were 5.6 +/- 1.6 (mean +/- SD) and 26 +/- 4 mmol/kg wet wt, respectively. In normal heart, [ATP] was 5.8 +/- 1.6 and [PCr] was 10 +/- 2 mmol/kg wet wt. These values are in excellent agreement with prior NMR studies and biopsy data. The protocol is easily accommodated within existing 1D cardiac patient protocols, and the same approach is advantageous for eliminating tissue volumetry and sensitivity corrections when measuring concentrations by 2D and 3D resolved spectroscopy. PMID- 8722818 TI - Single-shot diffusion MRI of human brain on a conventional clinical instrument. AB - A single-shot diffusion MRI technique on a standard clinical 1.5T scanner is presented. The method incorporates the following elements: (a) an inversion RF pulse followed by a delay of 1.3 s to null cerebral spinal fluid (CSF) signal, (b) a stimulated echo sequence (TE = 56 ms, TM = 100 ms) to obtain strong diffusion weighting, (c) a single-shot gradient- and spin-echo (GRASE) sequence for imaging with a modified k-space trajectory and Carr-Purcell Meiboom-Gill (CPMG)-phase cycle. The trace of the diffusion coefficient obtained with this approach is in good agreement with values reported for animal brain, and for recent human studies. It is demonstrated that single-shot diffusion imaging of human brain is feasible on an unmodified standard instrument without high gradient slew rate or extreme field homogeneity. PMID- 8722819 TI - Removal of lipid artifacts in 1H spectroscopic imaging by data extrapolation. AB - Proton MR spectroscopic imaging (MRSI) of human cerebral cortex is complicated by the presence of an intense signal from subcutaneous lipids, which, if not suppressed before Fourier reconstruction, causes ringing and signal contamination throughout the metabolite images as a result of limited k-space sampling. In this article, an improved reconstruction of the lipid region is obtained using the Papoulis-Gerchberg algorithm. This procedure makes use of the narrow-band-limited nature of the subcutaneous lipid signal to extrapolate to higher k-space values without alteration of the metabolite signal region. Using computer simulations and in vivo experimental studies, the implementation and performance of this algorithm were examined. This method was found to permit MRSI brain spectra to be obtained without applying any lipid suppression during data acquisition, at echo times of 50 ms and longer. When applied together with optimized acquisition methods, this provides an effective procedure for imaging metabolite distributions in cerebral cortical surface regions. PMID- 8722821 TI - NMR studies on water and polymer diffusion in dextran gels. Influence of potassium ions on microstructure formation and gelation mechanism. AB - At room temperature aqueous solutions of dextrans with concentrations > 25% (w/w) exhibit a sol-gel transition in the presence of > 1.0 M potassium chloride. In dextrans the gelation was unexpected due to missing anionic groups that usually provide the binding sites for cations. The quantitative investigation of the gel formation is based on changes of the diffusibility of water and dextran chains. The apparent diffusion coefficients of bulk water (in the order of 10(-6) cm2/s) and of water trapped in the junction zones as well as of polymer chains (in the order of 10(-7) to 10(-8) cm2/s) are determined by employing pulsed field gradient stimulated echo (PFGSTE) NMR. The restricted diffusion of bulk water in viscous sols and in soft and rigid gels has been quantitatively analyzed providing data for interbarrier distances (pore size), permeabilities of the diffusion barriers (density of junction zones) and interbarrier diffusion coefficients of water. Based on already published x-ray structure data and in accordance with the diffusion data presented in this paper "potassium-bonding" is assumed to be the most important interaction for the formation of a microstructure and for the stabilization of cross-links. The ionic radius of the potassium ion perfectly fits to the cage established by six oxygen atoms of glucose units of three polymer chains. Other cations, such as Li+, Na+, Rb+ and Cs+, according to their nonfitting ionic radii, do not provoke dextran gelation under these conditions. The mechanism of the transitions from sol to soft gel and further to rigid gel is discussed on the basis of restricted diffusion and x-ray structure data. PMID- 8722820 TI - Metabolite 1H relaxation in normal and hyponatremic brain. AB - Proton spin relaxation rate constants in normal and hyponatremic rat brain were measured to determine the sensitivity of metabolite relaxation properties to cytotoxic edema and to quantify metabolite concentration in normal and edematous brain. Relaxation rate constants for protons of water and spectral regions with dominant contributions from methyl protons of cholines (Cho), creatines (Cr), N acetylaspartate (NA), and lactate (Lac), and for methylene protons of glutamate (Glu) were measured at 7 T. Changes in metabolite relaxation properties associated with cytotoxic edema were a decrease in the Cr longitudinal rate constant, from 0.63 +/- 0.02 s-1 (mean +/- SE) in controls to 0.50 +/- 0.03 s-1 in edematous brain, and an increase in the transverse rate constant of NA from 5.3 +/- 0.2 s-1 in controls to 6.6 +/- 0.3 s-1 in edematous brain. Four hours after induction of hyponatremia, there was a 14% reduction in summed metabolite concentrations of Cho, Cr, and NA, and a 200% increase in Lac signal intensity. It is concluded that changes in both metabolite spin relaxation and detectable spin concentration accompany the cerebral pathology of cytotoxic edema complicated with secondary ischemia. PMID- 8722822 TI - MR measurement of relative water content and multicomponent T2 relaxation in human breast. AB - MR techniques providing accurate measurement of relative volumetric water content and multicomponent T2 relaxation times from a large volume of interest, have been implemented for characterization of breast tissue in vivo. In a sequence of 20-s breath-holds, data are obtained from much of the breast volume while suppressing signal from the chest wall and torso. Relative water content of each breast is calculated from one-dimensional fat and water profiles obtained using a hybrid two-point Dixon method (TE/TR = 17/5000 ms). Multicomponent T2 relaxation measurements are calculated from T2 decay curves obtained using a CPMG train of rectangular pulses (TE/TR = 8/5000 ms, 140 echoes) preceded by saturation pulses to localize longitudinal magnetization spatially. These techniques, validated in phantoms and human volunteers, are suitable for quantitative study of breast tissue in vivo, and in particular to investigate the potential role of MR for assessment of breast cancer risk. PMID- 8722824 TI - Signal-to-noise ratio comparison between surface coils and implanted coils. AB - The signal-to-noise ration (SNR) from implanted coils is widely known to be superior to that from surface coils. This article addresses the quantitative aspects of this improvement by explicitly evaluating the magnetic vector potential in a conducting medium of finite extent for both implanted and surface coils. The predictions of the model are tested with actual image data from spin warp experiments on gelatin phantoms. The authors derived a simplified expression that yields the gain in SNR of an implanted coil relative to that of a surface coil and is valid in many practical situations. PMID- 8722823 TI - Myocardial perfusion modeling using MRI. AB - In the present study, it is shown that it is possible to quantify myocardial perfusion using magnetic resonance imaging in combination with gadolinium diethylenetriaminopentaacetic acid (Gd-DTPA). Previously, a simple model and method for measuring myocardial perfusion using an inversion recovery turbo-FLASH (fast low-angle shot) sequence and Gd-DTPA has been presented. Here, an extension of the model is presented taking into account fast and slow water exchange between the compartments, enabling the calculation of the unidirectional influx constant (Ki) for Gd-DTPA, the distribution volume of Gd-DTPA (lambda), the vascular blood volume (Vb), and the time delay through the coronary arteries (delta T). The model was evaluated by computer simulation and used on experimental results from seven healthy subjects. The results in the healthy volunteers for a region of interest placed in the anterior myocardial wall were (mean +/- SD) Ki = 54 +/- 10 ml/100 g/min, lambda = 30 +/- 3 ml/100 g, Vb = 9 +/- 2 ml/100 g, delta T = 3.2 +/- 1.1 s. These results are in good agreement with similar results obtained by other methods. PMID- 8722825 TI - A real-time reconstruction system for magnetic resonance imaging. AB - A digital-electronic reconstruction system for MRI has been designed and demonstrated. The system is capable of reconstructing a 128 x 128 pixel image from complex-valued data in approximately 8 ms (122 frames per second) or a 256 x 256 pixel image in 32 ms (30 frames per second) using the standard 2D FFT reconstruction algorithm. Real-time MR imaging can be obtained when this reconstruction system is coupled with fast continuous echo-planar type data acquisition. This provides the unique potential for real-time monitoring of interventional procedures or for rapid patient positioning. The real-time reconstruction system presented here consists of four main subsystems: an analog to digital converter, an interface memory, the Fourier processor, and the display processor. The basic design of this reconstruction system is presented along with results, demonstrating the capability of the system. PMID- 8722826 TI - Hemodynamically independent analysis of cerebrospinal fluid and brain motion observed with dynamic phase contrast MRI. AB - Brain and cerebrospinal fluid (CSF) movements are influenced by the anatomy and mechanical properties of intracranial tissues, as well as by the waveforms of driving vascular pulsations. The authors analyze these movements so that the purely hemodynamic factors are removed and the underlying mechanical couplings between brain, CSF, and the vasculature are characterized in global fashion. These measurements were used to calculate a set of impulse response functions or modulation transfer functions, characterizing global aspects of the vasculature's mechanical coupling to the intracranial tissues, the cervical CSF, and the cervical spinal cord. These functions showed that a sudden influx of blood into the head was rapidly accommodated by some type of intracranial reserve or capacity. After this initial response, an equal volume of CSF was driven through the foramen magnum over the next 200-300 ms as the intracranial reserve relaxed to its base-line state. PMID- 8722827 TI - Frequency response of multi-phase segmented k-space phase-contrast. AB - A theoretical analysis of the temporal frequency response of multi-phase segmented k-space phase-contrast was developed. This includes the effects of both segment duration and the number of cardiac phases that are reconstructed. An increase in the number of views per segment and the corresponding increase in segment duration results in an increased smoothing or low-pass filtering of the time-resolved flow waveform. Reconstruction of all intermediate cardiac phases makes the Nyquist sampling frequency independent of the number of views per segment. This analysis was verified experimentally using a multi-phase phase contrast segmented k-space MR pulse sequence. This sequence reconstructs all intermediate cardiac phases and uses fractional segments at the end of the cardiac cycle if an entire segment does not fit. The use of fractional segments increases the portion of the cardiac cycle over which data are acquired. PMID- 8722828 TI - Diffusion-weighted interleaved echo-planar imaging with a pair of orthogonal navigator echoes. AB - This work describes a diffusion-weighted (DW) interleaved echo-planar imaging (IEPI) method for use on either conventional whole-body scanners or scanners equipped with high-speed gradient and receiver hardware. In combination with cardiac gating and motion correction with a pair of orthogonal navigator echoes, the presented method is time-efficient, compensates for patient motions, and is less sensitive to image distortions than single-shot methods. The motion correction scheme consists of correction for constant and linear phase terms found from the orthogonal navigator echoes. The correction for the linear phase term in the phase-encode direction includes gridding the data to the Cartesian grid. The DW IEPI was used to image a phantom rotating about the slice-select direction, and motion correction was performed to eliminate ghost artifacts arising from motion in either the readout- or phase-encoding directions. DW IEPI with motion correction was performed on a normal volunteer and on a patient with a 26-day-old region of ischemia over much of the right hemisphere. PMID- 8722829 TI - Myocardial perfusion and function in dogs with moderate coronary stenosis. AB - MRI studies of first-pass contrast enhancement with polylysine-Gd-DTPA and myocardial tagging using spatial modulation of magnetization (SPAMM) were performed to assess the feasibility of a combined regional myocardial blood flow and 2D deformation exam. Instrumented closed-chest dogs were imaged at a baseline control state (Cntl) followed by two interventions: moderate coronary stenosis (St) achieved by partial occlusion of the left anterior descending (LAD) and moderate coronary stenosis with dobutamine loading (StD). Hypoperfusion of the anterior region (ANT) of the myocardium (LAD distribution) relative to the posterior wall (POS) based on the upslope of the signal intensity time curve from the contrast-enhanced MR images was demonstrated only with dobutamine loading (ANT:POS Cntl = 1.077 +/- 0.15 versus ANT:POS StD = 0.477 +/- 0.11, P < 0.03) and was confirmed with radiolabeled microspheres measurements (ANT:POS Cntl = 1.18 +/ 0.2 ml/min/g versus ANT:POS StD = 0.44 +/- 0.1 ml/min/g; P < 0.002). Significant changes in regional myocardial shortening were only seen in the StD state (P < 0.02); the anterior region showed impaired myocardial shortening with dobutamine loading (P = NS), whereas the nonaffected POS region showed a marked increase in shortening when compared with Cntl (Cntl = 0.964 +/- 0.02 versus StD = 0.884 +/- 0.03; P < 0.001). These results demonstrate that an integrated quantitative assessment of regional myocardial function and semiquantitative assessment of myocardial blood flow can be performed noninvasively with ultrafast MRI. PMID- 8722830 TI - T1 rho relaxation and its application to MR histology. AB - The application of T1 rho an an alternative contrast parameter in high-field magnetic resonance histology (MRH) has been investigated. Spectroscopic measurements of T1 rho were performed on 5.75% agar and 1.0 mM MnCI2 phantoms at 9.4 T to validate the accuracy of the imaging measurements. Image studies were performed at 2.0 and 9.4 T on perfusion-fixed 17.5-day-old mouse embryos. T1, T2, and T1 rho relaxation times were calculated for the phantoms and muscle, diencephalon, and liver tissues. The 5.75% agar phantom and all tissues showed T1 rho dispersion with B1L, whereas the 1.0 mM MnCI2 phantom showed no significant B1L dependence. T1 rho dispersion with B(O) was observed arising from the effects of diffusion through susceptibility-induced gradients. T1 rho shows promise as a contrast parameter in high-field MRH because it is capable of producing T2-like contrast without the susceptibility artifacts associated with T2-weighted images. PMID- 8722831 TI - Evaluation of methemoglobin as an autologous intravascular MRI contrast agent. AB - Methemoglobin (MetHb) was evaluated as an intravascular paramagnetic contrast agent. Methemoglobin formation was induced by 4-dimethylaminophenol (4-DMAP), causing a reduction in blood T2* in vitro. The 4-DMAP generated metHb with a time constant of 62 s. A 4-DMAP bolus did not decrease measurably the signal intensity in the in vivo rabbit kidney in the first pass. At steady state, a MetHb concentration of 24.8 +/- 2.3% resulted in a signal decrease of 9.2 +/- 2.6% in the kidney. Methemoglobin is an effective vascular T2* relaxation agent, but the formation of MetHb by 4-DMAP is too slow for first-pass imaging. A more effective conversion agent resulting in a bolus of at least 25% MetHb within 5 s would result in a detectable first-pass signal and a viable contrast technique. PMID- 8722832 TI - Improved MR images of arterial specimens by submersion in trichlorotrifluoroethane. AB - MR images of ex vivo arterial specimens immersed in 1,1,2-trichloro-1,2,2 trifluoroethane (R-113) have improved signal-to-noise ratio and contrast-to-noise ratio. R-113 has no hydrogen atoms, so it yields no proton signal; hence, the contrast between the specimen and its background is maximized. SNR is maximized because (i) R-113 is nonconductive so that coil loading and inductive noise are minimized, and (ii) the volume susceptibility of R-113 closely matches that of water and tissue so that T2* effects are minimized. Short-term submersion of porcine aortas in R-113 was found to have no significant effect on the artery's hydration level, relaxation time, tensile strength, and structure or quantity of elastin, collagen, or smooth muscle cells. PMID- 8722833 TI - The role of the gastrointestinal tract in hematogenous candidiasis: from the laboratory to the bedside. AB - The gastrointestinal (GI) tract is a frequent source of hematogenous candidiasis in humans. Animal models of GI and hematogenous candidiasis have provided insights into the nature of candidal infection of host mucosal tissue, mechanisms of fungal dissemination to body organs, and features of host response to candidal infections. Biological systems such as these that simulate human candidiasis can be used for testing novel antifungal drugs. We have focused on two murine models of candidiasis with similarities to this fungal disease in humans. The first model simulates a commensal association of Candida albicans with the GI tract of immunocompetent hosts; it has permitted studies of innate and immune cell response to long-term ( > 60 days) infection of the esophageal, gastric, and intestinal mucosa. The second model simulates candidal infection in granulocytopenic patients with invasive candidiasis that originated from sites of colonization in the gut. Both models are well suited for investigating new approaches to prevention and treatment of hematogenous candidiasis. A review of the data on the role of GI candidiasis in hematogenous candidal infections is presented. PMID- 8722834 TI - Nosocomial candidiasis: emerging species, reservoirs, and modes of transmission. AB - During the 1980s, the frequency of nosocomial candidiasis increased dramatically. This trend has continued into the 1990s, and Candida species remain a major cause of nosocomial infections. Although Candida albicans remains the most frequent cause of fungemia and hematogenously disseminated candidiasis, a number of reports have documented infections caused by other Candida species: C. tropicalis, C. glabrata, C. parapsilosis, C. krusei, and C. lusitaniae. Many of these infections arise from an endogenous source, and their frequency is influenced by the patient population, the various treatment regimens, and the antibiotics or other supportive care measures employed at specific institutions. Additional infections may be accounted for by exogenous acquisition via the hands of health care workers, contaminated infusates and biomaterials, and the inanimate environment. Ongoing investigation should help improve our understanding of the epidemiology of candidiasis and facilitate the development of rational preventive measures. PMID- 8722835 TI - Problems and controversies in the management of hematogenous candidiasis. AB - Hematogenous candidiasis is associated with substantial mortality and morbidity. Amphotericin B has routinely been used to treat this infection. However, tolerance of therapy with amphotericin B is limited by the drug's toxicity. The results of recently completed prospective randomized clinical studies comparing amphotericin B with fluconazole for the treatment of hematogenous candididiasis suggest that fluconazole is as effective as amphotericin B and that fluconazole is better tolerated by patients. Nevertheless, several questions remain to be answered regarding the optimal choice of antifungal agent for both nonneutropenic and neutropenic patients, the dosing schedule and duration of therapy, the role of combination antifungal therapy, and the efficacy of the lipid formulations of polyenes. Controversial issues with respect to the role of central venous catheters in the pathogenesis of hematogenous candidiasis, as well as the roles of cytokines and white blood cell transfusions in the treatment of neutropenic patients with hematogenous candidiasis, also need to be addressed. PMID- 8722836 TI - Histoplasmosis and blastomycosis. AB - Histoplasmosis and blastomycosis are caused by dimorphic fungi, can be epidemic or endemic, and can produce a spectrum of illness, from subclinical infection to progressive disseminated disease. Diagnosis of both is best made by visualization of yeast in tissue or by culture. Itraconazole is the drug of choice for treatment of both histoplasmosis and blastomycosis, except in cases of life threatening infection, for which amphotericin B is indicated. A heavy inoculum of Histoplasma capsulatum may cause acute pulmonary infection in an otherwise healthy host, resulting in fever, hypoxia, and pulmonary infiltrates. Opportunistic histoplasmosis develops as chronic pulmonary histoplasmosis in those with a structural defect in the lung (emphysema) or as disseminated histoplasmosis in patients with cellular immune deficiency (due to immunosuppressants or AIDS). Blastomyces dermatitidis causes both pulmonary and extrapulmonary disease. Lung involvement may mimic bacterial pneumonia, while chronic presentations mimic lung cancer or tuberculosis. Skin is the most common extrapulmonary site of disease, followed by bone, prostate, and central nervous system. PMID- 8722837 TI - The new fungal opportunists are coming. AB - Frequent and prolonged exposure of immunocompromised patients to a variety of environmental conditions has resulted in the recognition of infections with new fungal opportunists. Infections with fungi such as Fusarium species, Paecilomyces lilacinus, Acremonium species, Trichosporon beigelii, Blastoschizomyces capitatus, Malassezia furfur, Penicillium marneffei, Scedosporium prolificans, and other dematiaceous species have become significant problems in the treatment of immunocompromised hosts. This discussion addresses several issues of pathogenesis, epidemiology, diagnosis, and treatment with regard to these new opportunists, through a review of both general and specific concepts. The growing need for training in clinical mycology is also summarized. PMID- 8722838 TI - Recent advances in the management of cryptococcal meningitis in patients with AIDS. AB - The optimum regimen for the treatment of cryptococcal meningitis in patients with AIDS is still not totally clear. The triazoles fluconazole and itraconazole are associated with response rates of 50%-60%. Amphotericin B appears to be associated with a more rapid clearance of organisms, and there are some data suggesting that initial therapy with amphotericin B is preferable to that with triazoles; however, this finding has not been definitively shown in large comparative trials. Results of a recently completed large trial suggested that initial treatment with amphotericin B followed by triazole therapy is associated with an acute mortality rate (approximately 6%) that is substantially less than that in previous studies. Relapse is common (20%-60% of cases) if the patient does not receive chronic suppressive therapy. The drug of choice for maintenance therapy is fluconazole (200 mg/d). A recent trial showed that fluconazole was superior to itraconazole (200 mg/d) as suppressive therapy. Prophylactic use of fluconazole (200 mg/d) significantly decreases the incidence of cryptococcosis and mucosal candidiasis, especially in patients with CD4 cell counts of < 50/mm3. However, because of the lack of a survival benefit and the risk of the selection effect on fluconazole-resistant Candida, it is difficult to make the recommendation of routine prophylaxis with fluconazole for all patients with AIDS; the decision to use prophylaxis should be based on more selective criteria. PMID- 8722839 TI - Is vulvovaginal candidiasis an AIDS-related illness? AB - Early in the AIDS epidemic, retrospective studies reported that vaginal candidiasis occurred more frequently in women who were infected with human immunodeficiency virus (HIV) than in those who were not infected. Some investigators suggested that new onset or recurrent vaginal candidiasis might identify HIV-infected individuals and predict the course of AIDS in women already known to be infected. In this article, studies of vaginal candidiasis in HIV infected women are examined, and several observations are made. First, early studies were small and likely reflected biased populations. Second, adherence to previously accepted diagnostic criteria for vaginal candidiasis was not consistent in these studies. Finally, conclusions about the increased risk of recurrent or chronic candidal vaginitis in HIV-infected women have been promulgated in the medical literature and may have influenced clinical practice even though such statements are not supported epidemiologically. Prospective trials with uninfected community controls should determine the true impact of HIV infection on vulvovaginal candidiasis. PMID- 8722840 TI - Common superficial fungal infections in patients with AIDS. AB - Superficial mycotic infections such as seborrheic dermatitis, tinea pedis, tinea corporis, and onychomycosis are common in patients infected with human immunodeficiency virus (HIV). In communities where HIV infections are frequent, some of these clinical presentations serve as markers of the stage of HIV infection. The diagnosis of superficial fungal infection in HIV-positive patients may be difficult because of atypical clinical manifestations. Therefore, to ensure a correct diagnosis, skin scrapings should be collected for potassium hydroxide preparations and cultures. Most forms of dermatophytosis in HIV positive patients respond well to many topical antifungal agents, such as azoles, terbinafine, and ciclopirox olamine. If the disease is chronic and extensive, then ketoconazole, fluconazole, and itraconazole are each effective. PMID- 8722841 TI - Lipid formulations of amphotericin B: recent progress and future directions. AB - Three lipid formulations of amphotericin B are now either marketed for clinical use or undergoing further study before they can be approved in various countries worldwide. Amphotericin B lipid complex (ABLC; Abelcet, Liposome Company, Princeton, NJ) is a concentration of ribbonlike structures of a bilayered membrane formed by combining a 7:3 molar ratio of dimyristoyl phosphatidylcholine and dimyristoyl phosphatidylglycerol with amphotericin B. Amphotericin B colloidal dispersion (ABCD; Amphocii, Sequus Pharmaceuticals, Menlo Park, CA) is composed of disklike structures of cholesteryl sulfate complexed with amphotericin B. AmBisome (Nexstar, San Dimas, CA), the only true liposomal amphotericin B, consists of small unilamellar vesicles made up of a bilayer membrane of hydrogenated soy phosphatidylcholine and distearoylphosphatidylglycerol stabilized with cholesterol in a 2:0.8:1 ratio combined with amphotericin B. All of the preparations appear to be preferentially accumulated in organs of the reticuloendothelial system, as opposed to the kidney. In vivo animal models as well as current clinical experience suggest that use of these formulations results in overall improvement in the therapeutic index. Patients with life-threatening mycosis for whom therapy has failed or who are intolerant to therapy with amphotericin B deoxycholate have been successfully treated with these formulations. However, further study is warranted to help clarify the usefulness of each of the lipid formulations as first-line therapy for documented or suspected invasive fungal infections. PMID- 8722842 TI - Amphotericin B: a commentary on its role as an antifungal agent and as a comparative agent in clinical trials. PMID- 8722843 TI - Role of azoles in antifungal therapy. AB - The addition of itraconazole and fluconazole to the antifungal armamentarium has resulted in less toxic, easily administered therapy for both endemic and opportunistic mycoses. Itraconazole has become the treatment of choice for most patients with mild-to-moderate histoplasmosis, blastomycosis, and sporotrichosis. Coccidioidomycosis is effectively treated with either itraconazole or fluconazole; fluconazole has become the drug of choice for the treatment of coccidioidal meningitis. In several large cooperative trials, fluconazole has been shown to have a useful role in the treatment of patients with candidemia, localized forms of mucocutaneous and visceral candidiasis, and cryptococcal meningitis. PMID- 8722844 TI - Comparison of the efficacy of amphotericin B and fluconazole in the treatment of cryptococcosis in human immunodeficiency virus-negative patients: retrospective analysis of 83 cases. French Cryptococcosis Study Group. AB - We retrospectively analyzed clinical outcome of meningeal and extrameningeal cryptococcosis in HIV-negative patients treated with amphotericin B (43 patients) or fluconazole (40 patients). Amphotericin B and fluconazole were prescribed equally to patients with neoplastic diseases and no risk factor, but organ transplant recipients and patients with other diseases were mostly given fluconazole and amphotericin B, respectively. Patients with more severe infections (i.e., meningitis, neurological disorders, or higher levels of antigen in cerebrospinal fluid) were more frequently treated with amphotericin B. A cure rate of > 70% was achieved regardless of the initial treatment and the severity of the infection. A Cox regression analysis showed that age of > 60 years, neoplastic disease, abnormal mental status, disseminated infection at the time of diagnosis, and therapeutic failure were independent predictors of death. Although fluconazole appears to be as effective as amphotericin B, only a prospective multicenter study will determine the best treatment regimen for patients with cryptococcal meningitis who do not have AIDS. PMID- 8722845 TI - Is antifungal susceptibility testing useful in guiding fluconazole therapy? AB - Recently, the National Committee for Clinical Laboratory Standards (NCCLS) subcommittee on antifungal susceptibility testing published a proposed standard for a broth macrodilution method for in vitro susceptibility testing of yeasts. The major reason for developing such a method is to predict the likely clinical and microbiological outcome of treatment. A number of studies of fluconazole have attempted to correlate in vitro minimum inhibitory concentrations (MICs) with in vivo efficacy in humans by means of a variety of methods, including the NCCLS proposed standard. These studies have clearly demonstrated that the ability to predict clinical outcome with MICs is dependent on the patient population studies. Thus, fluconazole MICs were correlated with clinical outcomes for patients with AIDS-associated oropharyngeal candidiasis. In contrast, fluconazole MICs did not correlate with clinical outcomes for patients with candidemia who did not have AIDS; these patients have numerous factors that can independently influence the response to antifungal therapy aside from the MIC. Data on patients with AIDS-associated cryptococcal meningitis from my laboratory demonstrated that the use of fluconazole MICs in conjunction with clinical variables, such as blood culture results, enhances the ability to predict treatment outcome. More investigations addressing the role of host factors in the vitro-in vivo correlation are necessary before antifungal testing can be useful in guiding therapy. PMID- 8722846 TI - The future of antifungal therapy. AB - In the late 1970s the options for systemic antifungal therapy doubled with the addition of intravenous miconazole and oral ketoconazole to the two previously available agents, amphotericin B and flucytosine. The 1980s ushered in the next generation of triazole antifungals, fluconazole and itraconazole. These are the present-day mainstays of treatment for some of the most serious systemic fungal infections. However, the increase in the numbers and types of fungal pathogens, and especially the emergence of azole-resistant fungi, have prompted a continuing search for new therapeutic options. This search has yielded more-potent triazole antifungals, new vehicles for both polyenes and triazoles, and entirely new classes of agents such as the echinocandin derivatives; in addition, it has prompted the evaluation of new combinations of present-day antifungals and exploration of the use of immunomodulators for treatment of fungal infections. Rapid developments in molecular mycology are permitting a concentrated search for more targets for antifungals. We are entering a new era of antifungal therapy in which we will continue to be challenged by systemic fungal diseases but will have greatly expanded options for treatment. PMID- 8722847 TI - Identification of dematiaceous fungi and their role in human disease. AB - Phaeohyphomycosis is the disease caused by the dematiaceous hyphomycetes or those fungi that are darkly pigmented because of the presence of melanin in their cell wall. Infections with these fungi may remain localized at the site of traumatic inoculation or within the sinuses or may become disseminated. As these fungi pose a significant problem to both the clinician who must treat the patient and the laboratorian faced with identification of the organism, a selection of representative organisms was presented at the meeting entitled "Focus on Fungus Infections." These fungi often infect patients who are not overtly immunocompromised and therefore may be encountered in a wide variety of cases. PMID- 8722848 TI - Percutaneous Inoue-balloon valvuloplasty in patients with mitral stenosis and associated moderate mitral regurgitation. AB - The present study compares the outcome of percutaneous Inoue-balloon mitral valvuloplasty performed in 21 patients with (group A) and 83 patients without (group B) preexisting moderate mitral regurgitation, using our height-derived balloon-sizing method. All procedures were successfully completed without untoward complications. The immediate increments in mitral valve area measured by echocardiographic methods and optimal valvuloplasty results were significantly higher in group B compared with group A (0.9 cm2 vs. 0.7 cm2, P = 0.01, and 99% vs. 90%, P = 0.007, respectively). No patients in either group sustained a final grade > or = 3/4 angiographic mitral regurgitation. A mild increase in mitral regurgitation was encountered more commonly in group B than in group A patients (22% vs. 0%, P = 0.03). At a mean follow-up of 19-20 months, the substantial majority of patients (> or = 90%) in both groups continued to experience maintained symptomatic benefits. In conclusion, our preliminary data seem to indicate that percutaneous Inoue-balloon mitral valvuloplasty using our height derived balloon-sizing method in the stepwise dilatation approach in selected patients with significant mitral stenosis and concomitant moderate mitral regurgitation is associated with a low risk of developing severe mitral regurgitation, with effective mitral valve enlargement, and with sustained midterm symptomatic benefits. PMID- 8722850 TI - Advantage of Inoue balloon catheter in mitral balloon valvotomy: experience with 220 consecutive patients. AB - Percutaneous mitral balloon valvotomy (PMV) using the Inoue balloon catheter was attempted in 220 consecutive patients with severe symptomatic mitral stenosis. Their age range was 10-63 mean 30 +/- 10 years; 161 patients were females and 59 were males; 29 patients were in atrial fibrillation. Eleven patients were pregnant; 14 patients underwent previous surgical commissurotomy. The procedure was technically successfully performed in 215 (97.7%) patients. The mean fluoroscopy time was 15.5 +/- 6.4 min and mean procedure time was 109 +/- 79 min. Optimal results (group I) was achieved in 207 patients who have mitral score of 7 +/- 1. PMV resulted in decrease in left atrial pressure from 23 +/- 5 to 14 +/- 4 mm Hg (P < 0.001), the mean mitral valve gradient (MVG) decreased from 15 +/- 4 to 6 +/- 3 mm Hg (P < 0.001). The mitral valve area (MVA) by catheter increased from 0.7 +/- 0.2 to 1.7 +/- 0.5 cm2 (P < 0.001) and MVA as determined by echocardiography (2DE) increased from 0.8 +/- 0.1 to 1.9 +/- 0.3 cm2 (P < 0.001). The results were suboptimal in eight patients who have a mitral score of 10 +/- 1 (group II) MVA by catheter increased from 0.7 +/- 0.2 to 1 +/- 0.1 cm2 and Doppler MVA increased from 0.8 +/- 0.1 to 1.3 +/- 0.1 cm2. There were no deaths or thromboembolism. Two patients developed cardiac tamponade; mild mitral regurgitation (MR) developed in 24 patients (11%) and increased by one grade in another 22 patients (10%). Severe MR was encountered in three patients (1.4%). A small ASD (QP/QS < or = 1.3) was detected by oximetry in 5% of patients and by color-flow mapping in 26% of patients. One hundred fifty-eight patients from group I were followed up, for a mean of 32 +/- 12 months; MVA remained at 1.7 +/- 0.4 cm2. Seven patients developed mitral restenosis in group I, and 5 out of 8 patients developed restenosis in group II. We conclude that the hemodynamic results are good and comparable with those reported with double balloon technique. However, the Inoue balloon has several advantages over the double balloon technique: (1) low incidence of mitral regurgitation and ASDs; (2) shorter procedure and fluoroscopy time; and (3) low complication rates and the valve anatomy affects the immediate and late outcome of mitral balloon valvotomy. PMID- 8722851 TI - Top 10 reasons to use the Inoue balloon. PMID- 8722852 TI - Stent implantation for relief of pulmonary artery stenosis: immediate and short term results. AB - Our objective was to assess the immediate and short-term results of stent implantation to relieve pulmonary artery stenosis (PAS). Thirty-seven patients underwent an attempt at stent implantation at a median age of 7.0 years (range, 0.8-31.4 years) and a median weight of 20.5 kg (range, 7.4-85 kg). Twenty-two patients had previous tetralogy of Fallot repair. A total of 55 stents were implanted successfully in 36 patients. The peak systolic gradient across the stenotic segment decreased from a mean of 43 +/- 20.4 mmHg prestent to 13 +/- 13.9 mmHg (P < 0.001) poststent. The diameter of the narrowest segment increased from a mean of 4.8 +/- 1.6 mm to 10.5 +/- 2.6 mm (P < 0.001). The right ventricular-to-aortic mean systolic pressure ratio decreased from 0.74 +/- 0.2 to 0.52 +/- 0.19 (P < 0.001). Complications included balloon rupture prior to full stent expansion in 4 patients (in 2 patients the stent was positioned in the superior vena cava, and in 2 in the inferior vena cava), distal migration of a stent which was successfully retrieved at surgery 1 mo later in 1 patient, and tethering of the stent to the balloon requiring surgical removal in 1 patient. One patient died several hours after stent placement. Sixteen patients underwent repeat catheterization at a mean follow-up interval of 0.9 +/- 0.5 years (range, 0.2-2.0 years). The mean gradient across the stent for these 16 patients was 26.7 +/- 19.8 mmHg, and there was no change in the mean diameter (9.4 +/- 3.2 mm). Two patients developed stenosis related to neointimal proliferation at the stent site which was redilated successfully. In conclusion, stent implantation is generally safe and effective in relieving PAS. PMID- 8722853 TI - The superiority of stenting stenotic pulmonary arteries--best but not easiest. PMID- 8722854 TI - Tortuosity of coronary arteries in chronic pressure and volume overload. AB - The role of coronary tortuosity in the pathophysiology of chronic pressure and volume overload is still unclear. A new method for measuring coronary tortuosity in patients with chronic pressure and volume overload was evaluated in 62 patients. Sixteen controls, 14 patients with arterial hypertension, and 32 patients with aortic regurgitation were included in the present analysis. The left anterior descending (LAD) and circumflex (LCX) coronary arteries were traced, and tortuosity was determined in the 30 degrees right (RAO) and 60 degrees left (LAO) anterior oblique projection. Tortuosity index (TI, %) was defined as the percent ratio of calculated shortest distance divided by total length of the coronary artery. TI was 104.1 +/- 3.2% at end-diastole in controls, 105.7 +/- 3.8% in hypertensives (P < 0.05 vs. controls), and 102.9 +/- 2.5% in patients with aortic regurgitation (P < 0.05 vs. controls, P < 0.001 vs. hypertensives). Respective values at end-systole were 107.8 +/- 4.7% in controls, 109.8 +/- 7.1% in hypertensives (ns vs. controls), and 104.3 +/- 3.3% in patients with aortic regurgitation (P < 0.001 vs. controls and vs. hypertensives). No differences were found in tortuosity between RAO and LAO projection or between LAD and LCX artery. There was a significant correlation between TI and left ventricular (LV) muscle mass, LV volume, and age. Females tended to have more tortuous vessels than males. Coronary tortuosity is more pronounced in patients with chronic pressure than with volume overload. Determinants of coronary tortuosity are gender, age, LV volume, and muscle mass. Thus, coronary tortuosity seems to play an important role as a physiologic determinant for the flow and the mechanics of the vessel wall. PMID- 8722855 TI - Long-term intracoronary stent placement: arteriographic and histologic results after 7 years in a dog model. AB - Prosthetic intracoronary stenting has become an important adjunct to balloon angioplasty in the percutaneous treatment of coronary artery disease. However, there are few reports of the long-term histologic consequences of stenting in the world literature. We report the arteriographic, histologic, and ultrastructural examination of a flexible tantalum wire coil stent implanted in the coronary artery of a dog for 7 years. The vessel was arteriographically patent without evidence of either stenosis or ectasia. The stent had been incorporated into the arterial wall, and was covered with a neointima of approximately 100 microns maximum thickness. There was necrosis of the tunica media adjacent to the stent but the inflammatory response was restricted to occasional macrophages in the neointima of the immediate vicinity of the stent wire. The endothelial cell layer was normal in its morphologic appearance. In summary, the flexible tantalum wire coil stent was well tolerated by the host blood vessel and demonstrated acceptable biocompatibility for the 7-year duration of its implantation in the normal dog coronary artery. PMID- 8722856 TI - Porcine model of stent thrombosis: platelets are the primary component of acute stent closure. AB - Acute stent thrombosis remains a major concern of coronary stent implantation. Animal studies using stents do not adequately mimic this clinical problem, since stent placement is rarely associated with acute closure. The purpose of this study was to develop and characterize a porcine model of stent thrombosis. Improved understanding through such a model may be useful toward preventing and treating acute stent closure. Whole blood was drawn from domestic crossbred swine one day before study. Platelets were isolated, labeled with 111-In tropolone, and reinjected within 18 hr of the study. Bilateral carotid arteries were exposed, and severe injury induced by a series of mechanical crushes. This method produced histologic injury similar to human coronary angioplasty, with medial disruption and large dissections protruding into the lumen. Stenting was performed in standard fashion with 3.5-mm JJIS stents. Local platelet deposition was measured and recorded as 111-In radioactivity using a miniaturized scintillation detector (Dosimeter Corp.) mounted directly at the artery injury site. This measurement was made in real time at 1-min intervals. Similarly, volumetric blood flow was measured in real time by Doppler flowmeter. Eighteen arteries of nine pigs were studied. In nine arteries from nine pigs, crush injury only was performed and monitored. In the contralateral artery, crush injury was followed immediately by placement of a 3.5-mm Palmaz-Schatz (coronary) stent. Blood flow decreased rapidly following injury in both groups and followed a cyclic pattern. Eight arteries of the crush alone and two arteries of the crush plus stent groups were totally occluded 1 hr after crush. 111-In counts normalized to baseline were significantly higher at 1 hr in both groups compared to baseline; in the stented group, counts were higher than in the unstented group. Blood flow was higher in the stented group than in unstented group for 1 hr. Histopathologic observation of the thrombi forming in both crush-only and crush-stent injuries showed severe medial dissections with obstructing medial flap formation. The thrombi forming in both groups were highly platelet rich. This model of stent and arterial thrombosis showed rapid formation of platelet-rich thrombus, cyclic blood flow variations, and acute occlusion in 20% of cases. Stent placement at arterial injury sites is associated with thrombus that is predominantly platelet rich. Stent placement at injury sites enhances platelet deposition over crush injury alone. Despite greater numbers of platelets, as shown by increased 111-In counts, stenting improved vessel patency. These were likely due to higher volumetric blood flow, continuous deposition, and embolization of labeled platelets. PMID- 8722857 TI - Swine model of coronary restenosis: effect of a second injury. AB - Looking for a coronary artery restenosis model closer to human pathology, a protocol of balloon injury/reinjury (plaque of dilatation) in swine coronary artery was designed. Pig coronary arteries (n = 24) were dilated for this study: 12, group 1, once (sacrifice at 10.0 +/- 2.2 weeks); 6, group 2, twice at 2-wk intervals (sacrifice at 5.2 +/- 0.2 wk); 6, group 3, twice at 4-wk intervals (sacrifice at 9.3 +/- 1.9 wk). A single overdilatation resulted in an eccentric neointimal hyperplasia representing half of the wall area (group 1, 45.6 +/- 5.1%). In animals (groups 2 and 3) subjected to redilatation, fracture length, ratio of fracture length to internal elastic lamina (IEL) circumference, and neointimal hyperplasia response were similar to those observed in group 1. In group 3, the shape of the lesion appeared more concentric and the fracture of the IEL more fragmented than in group 1. Although this model of injury/reinjury did not lead to more severe intimal hyperplasia, performing a second angioplasty at the same site did lead to a more concentric intimal response, related to multiple fractures of the IEL. PMID- 8722858 TI - Comparison of tissue disruption caused by excimer and midinfrared lasers in clinical simulation. AB - Laser coronary angioplasty is a useful therapy for selected complex coronary lesions. Laser-induced acoustic trauma is postulated to be a cause of dissection and acute vessel occlusion. Controversy exists regarding the relative degree of photoacoustic effects of midinfrared and excimer lasers in clinical practice. To date, these systems have not been compared at clinical energy doses and with clinical pulsing strategies. Therefore, we studied the photoacoustic effects of both midinfrared and excimer lasing at clinically accepted doses. Human atherosclerotic iliofemoral artery segments were obtained at autopsy (n = 36) and placed lumen side up in a saline bath. Clinical laser catheters were advanced over an 0.018" guide wire, perpendicular to the tissue. A 10-g down force was applied to the catheter for full-thickness lasing. Pulsing strategies were, for midinfrared laser: 5 pulses, 1-sec pause, 5 pulses, 1-sec pause, 5 pulses, withdraw; for excimer: 5 sec of pulses, wait 10 sec, 5 sec of pulses. Several clinically acceptable energy levels were used; for excimer: 25 mJ/mm2, 40 mJ/mm2, 60 mJ/mm2; for midinfrared: 3 W (400 mJ/mm2), 3.5 W (467 mJ/mm2). Photoacoustic effect was assessed histologically by determining the number of lateral cleavage planes (dissections) arising from the lased crater border and extending into the surrounding tissue. In normal tissue, midinfrared lasing produced less acoustic damage than excimer lasing (2.79 +/- 0.78 vs. 5.27 +/- 0.75 cleavage planes, mean +/- SD, P < 0.05, data for lowest energy for each system). The same was true in noncalcified atheroma (2.48 +/- 0.71 vs. 6.43 +/- 1.09, P < 0.05) and calcified atheroma (2.47 +/- 1.21 vs. 6.27 +/- 1.13, P < 0.05). This effect was similar at all energy levels, with a trend for more damage at higher energies in both systems. This study demonstrates that midinfrared lasing causes less acoustic damage than excimer lasing when using clinical catheters, energy levels, and pulsing strategies. This effect is independent of tissue-type but tends to be dose-related. These findings may explain, in part, the differences in dissection rates seen clinically. PMID- 8722859 TI - Hapten resistant dextran induced ARDS reaction following intracoronary stent deployment. AB - A case of dextran induced adult respiratory distress syndrome is used to illustrate the most serious complication of dextran usage during intracoronary stent placement. This reaction occurred despite premedication with the hapten dextran 1. Mechanisms of dextran induced complications are discussed. A mechanism for hapten resistant dextran reactions is postulated. Suggestions to prevent such reactions are made. PMID- 8722860 TI - Spontaneous coronary dissection resulting in acute myocardial infarction: successful treatment with primary angioplasty. AB - Spontaneous coronary dissection is an unusual cause of acute myocardial infarction for which the optimal therapy has not been established. We report the case of a 63-year-old man with acute lateral wall myocardial infarction resulting from spontaneous dissection of the major diagonal branch of the left anterior descending artery in whom acute and long-term patency was restored by primary PTCA. PMID- 8722861 TI - Interventional cardiac catheterization therapy for combined coarctation of the aorta and patent ductus arteriosus: successful outcome in two infants. AB - We report two infants each with coarctation of the aorta and small patent ductus arteriosus who had balloon angioplasty therapy for aortic obstruction at 3-6 months of age followed by anterograde placement of a single Gianturco coil to close the ductus at a separate catheterization. One infant developed recoarctation that resolved with repeat angioplasty prior to closure of the ductus. Follow-up evaluations have demonstrated a trivial aortic gradient, no aortic aneurysm, and no residual shunt. PMID- 8722863 TI - Acute cyanosis following balloon angioplasty of residual arch obstruction after the Norwood I operation. AB - A 7-month-old child with residual aortic arch obstruction following a modified Norwood I operation developed profound cyanosis with subsequent bradycardia and hypotension immediately after successful balloon angioplasty. This complication of aortic angioplasty is specific to patients with this physiology. The pathophysiology of the event is discussed. PMID- 8722864 TI - Aortocaval fistula complicating abdominal aortic aneurysm: case report and literature review. AB - Aortocaval fistula is a rare complication of abdominal aortic aneurysm involving less than 1% of all abdominal aortic aneurysms. The diagnosis is difficult, and preoperative identification can be challenging. We describe an interesting case of a 72-year-old man presenting with acute-onset dyspnea who was found to have an aortocaval fistula secondary to a large abdominal aortic aneurysm. The clinical features that exemplify aortocaval fistula are discussed and the literature reviewed. PMID- 8722865 TI - Coronary artery bypass graft pseudoaneurysm communicating with the right atrium: a case report and review. AB - We report a pseudoaneurysm of the right coronary artery bypass graft with fistulous drainage into the right atrium. This patient presented with an acute myocardial infarction in a different vascular territory. Cardiac catheterization led to the diagnosis of the pseudoaneurysm. A review of pseudoaneurysms of aortocoronary bypass grafts is presented. PMID- 8722867 TI - Gradual angioplasty and stent implantation to treat complete superior vena cava occlusion after Mustard procedure. AB - A 16-year-old male was admitted with complete occlusion of the superior vena cava pathway 14 years after Mustard procedure for transposition of the great arteries. From a left subclavian vein approach, the atretic vein segment was perforated using a straight guidewire, and was followed by sequential balloon dilation. Implantation of a 30 mm Palmaz-stent through a femoral vein approach resulted in a widely patent channel of the vena cava superior into the systemic venous atrium. PMID- 8722866 TI - Pseudoaneurysm causing partial obliteration of the left atrium: case report and review. AB - We describe a case of left ventricular pseudoaneurysm with subepicardial dissection onto the left atrial wall, mimicking a smooth left atrial mass, causing partial obliteration of the left atrial cavity. This patient presented with a transient ischemic attack, most likely resulting from a thromboembolic episode to the brain. Emergent surgery was successfully performed and the patient is doing well at 1-year follow-up, with no signs of left atrial compression by transthoracic echocardiography. PMID- 8722869 TI - Implantation of Palmaz stents in branch pulmonary arteries using Olbert balloons. AB - Twenty-three patients, ranging in weight between 4.9 and 74 kg, underwent stent implantation in branch pulmonary arteries. At first, high-pressure balloons were used for stent implantation but, because of stents slipping off the balloons, subsequently in eight patients, aged 2-18 years and weighing 11-72 kg, Olbert balloons were used. Fifteen stents were implanted in 11 vessels in these 8 patients. No instance of stent slippage occurred during introduction through the sheath or when the stent was passed around the curves of the right heart. In one patient, the fully expanded stent slipped over the wire into the main pulmonary artery. This occurred because of undersizing of the stent. The stent was re advanced into position and expanded using a larger balloon. No significant complications occurred with these balloons. The use of Olbert balloons for stent implantation in branch pulmonary arteries is recommended. PMID- 8722870 TI - Incomplete occlusion of left ventricular aneurysms after endoventricular aneurysmorrhaphy: diagnosis by echocardiography and ventriculography. AB - Surgical treatment of left ventricular aneurysms have recently focused on maintaining normal left ventricular geometry by using a circular patch repair to exclude the aneurysmal cavity (endoaneurysmorrhaphy). We describe two patients who underwent this procedure and were subsequently found by echocardiography and angiography to have a residual communication between the left ventricular cavity and the aneurysm which contained thrombus. This finding may have implications regarding the optimal hemodynamic result of the surgery and the risk of thromboembolism. PMID- 8722871 TI - Coronary flow after stenting. PMID- 8722872 TI - Selective catheterization of the anomalous left coronary artery arising from the pulmonary trunk. PMID- 8722873 TI - How to avoid having a stent mounted on a ruptured balloon in a coronary artery. PMID- 8722874 TI - Appropriate methods for bone fixation. PMID- 8722875 TI - Perioperative management of the rheumatoid patient. AB - Rheumatoid arthritis is a systemic disease that can affect multiple organ systems of the body. It is a symmetrical polyarthropathy which leads to destruction of cartilage and bone. Before undertaking surgery on the rheumatoid patient, several issues need to be addressed. A thorough evaluation by a multidisciplinary team is essential to decrease any operative risks involved. This is a review of some of the systemic manifestations of rheumatoid arthritis with an emphasis on anesthesiology, and radiological consultation, and perioperative surgical management of rheumatoid drug therapy. PMID- 8722876 TI - Arthroscopy of the calcaneocuboid and talonavicular joints. AB - The authors describe the anatomy, portal placement, technique, indications, and preliminary results of seven cases of calcaneocuboid or talonavicular joint pathology that underwent elective investigational diagnostic or therapeutic arthroscopy with a follow up of 3-14 months. All patients had failed available conservative therapy. Results of treatment were five excellent and two good cases. Two patients required arthrotomy. There were no complications associated with portal placement, arthroscopic technique, or instrumentation. Arthroscopy of the calcaneocuboid and talonavicular joints has proven to be a relatively safe and valuable diagnostic and therapeutic tool. It is hoped that advances in small joint arthroscopy application will encourage further technological evolution in arthroscopic instrumentation and establish a foundation for further arthroscopic exploration of the small joints of the foot. PMID- 8722877 TI - Long survival with metastatic clear cell sarcoma of the Achilles tendon. AB - Clear cell sarcoma of tendons and aponeuroses is a rare tumor with a predilection for the lower extremities, particularly the foot. Although it is claimed to have a poor prognostic outlook, it is sometimes characterized by a slow clinical course. The authors report a case with a prolonged survival period of 7 years--5 years in the presence of multiple lung metastases. PMID- 8722878 TI - Morton's interdigital neuroma: a clinical review of its etiology, treatment, and results. AB - Morton's neuroma is most likely a mechanically induced degenerative neuropathy which has a strong predilection for the third common digital nerve in middle-aged women. The excessive motion between the third and fourth metatarsals, the tethered third common digital nerve in the third web space, the third and fourth metatarsal heads flanking the third common digital nerve, the stout third transverse intermetatarsal ligament overlying the third common digital nerve, and excessive weightbearing stress on the forefoot, particularly by wearing pointed and high-heeled shoes, can collectively produce microdamage to the third common digital nerve. If allowed to continue for a long period of time, this can become manifested microscopically by nerve fiber degeneration and excessive intraneural and juxtaneural reparative fibrous tissue formation resulting in a significantly enlarged nerve. Such enlargement can create further trauma, and therefore become even more symptomatic. When nonsurgical means fail to relieve patient's symptoms, surgical removal of this offending neuroma through a dorsal approach can produce dramatic relief of symptoms. In addition, when a painful recurring Morton's neuroma does not respond to conservative treatments, removal of this lesion through a plantar approach can provide lasting relief. PMID- 8722879 TI - Ankle and subtalar fusion utilizing a tricortical bone graft, bone stimulator, and external fixator after avascular necrosis of the talus. AB - A unique ankle and subtalar fusion was performed on a patient with avascular necrosis of the talus. The body of the talus was removed and replaced with an autogenous tricortical bone graft. Healing was aided by the use of a bone stimulator and external fixator. The authors believe this fusion to be a more stable fusion than others used to treat avascular necrosis of the talus. PMID- 8722880 TI - The Tillaux fracture. AB - The Tillaux fracture is an avulsion fracture of the distal anterior tibial tubercle that is seen in adolescents. The authors present a literature review that includes classification, mechanism of injury, epiphyseal development, and treatment. Two cases of the Tillaux fracture are presented that represent patients of different physeal maturity and severity of injury, and different techniques of osteosynthesis. PMID- 8722881 TI - The surgical dilemma of the malunited calcaneal joint depression fracture: the VAMC Miami experience. AB - During the period of October 1985 through March 1995, 25 patients presented to the Veterans Affairs Medical Center, Miami with a chief complaint of chronic pain status post calcaneal joint depression fracture. Their acute injury was treated conservatively at this and various other institutions. Nine patients were managed conservatively with orthotics, University of California Biomechanics Laboratory braces (UCBLs), shoe modification, or prosthetics. Sixteen patients underwent hindfoot fusions. Good to excellent results were obtained in 15 of the 16 fusions. The remaining patient, having an isolated subtalar fusion, was later diagnosed with multiple sclerosis. Proper evaluation of the chronic pain patient is critical when mapping out an appropriate treatment plan. Hindfoot arthrodesis with soft tissue decompression is a successful tool in eliminating chronic pain due to malunited depression fractures. PMID- 8722882 TI - Reconstruction of an Achilles tendon defect utilizing an Achilles tendon allograft. AB - Acute ruptures of the Achilles tendon are common. Treatment for such ruptures is well documented. Treatment for chronic ruptures with a significant defect poses a surgical dilemma. The authors present the use of a tendon allograft to replace an Achilles tendon defect with a 2.5-year clinical follow-up inclusive of isokinetic testing. PMID- 8722883 TI - A comparison of weightbearing pressures in various postoperative devices. AB - Currently, there are a multitude of computerized gait analysis systems to evaluate weightbearing patterns in the foot. The following study utilizes the F Scan System of foot pressure analysis. This system was applied to evaluate plantar pressures as they occur in various postoperative shoe gear and to determine which devices are most clinically beneficial in ambulatory surgery. A pressure sensor recorded plantar pressures in 20 feet wearing various types of postoperative shoe wear. Initially, a stockinette was used to simulate barefoot pressure, followed by a wooden postoperative shoe, a Darco postoperative shoe, a Darco closed postoperative shoe, a Darco "softie," a Darco wedge, an Equalizer Premium Walker, an Equalizer Low Top Walker, and an Aircast. The results demonstrated that the Equalizer Premium Walker was most efficient in reducing forefoot pressures, while the postoperative surgical shoes had variable results, noticeably with several increases in pressure being recorded. PMID- 8722884 TI - The dose effect of continuous passive motion in postoperative rehabilitation of the first metatarsophalangeal joint. AB - Continuous passive motion (CPM) has been utilized in postoperative rehabilitation of many joints of the body. Little has been published regarding its applications in the first metatarsal phalangeal joint or other joints of the foot. A review of the basic science and clinical studies regarding the dose effects on the healing of articular cartilage defects and periarticular tissues has led to possible correlations in treating disorders of the first metatarsal phalangeal joint. Proposed protocols are being presented which may be helpful in future clinical studies to confirm the benefits of CPM in this joint of the foot. PMID- 8722885 TI - Modified Elmslie lateral ankle stabilization procedure. AB - An alternative to traditional lateral ankle stabilization procedures is presented using freeze-dried fascia lata graft tissue. This procedure is minimally invasive and obviates the complications typically associated with tenodesis procedures. Furthermore, the fascia lata graft is more anatomically similar to ligament tissue, and studies suggest that the architecture is maintained through a neovascularization process. PMID- 8722886 TI - Talonavicular joint arthrodesis for paralytic post poliomyelitis forefoot instability. AB - A 17-year-old patient presented with paralytic valgus left foot deformity after poliomyelitis. Forefoot instability resulted from the transfer of peroneus brevis to the second metatarsal bone. This was secondary to a medial gapping of the talonavicular joint as a result of unbalanced pull of the transferred tendon. Talonavicular arthrodesis following a medial-based wedge articular resection has corrected the deformity. PMID- 8722887 TI - Bleomycin sulfate in the treatment of mosaic plantar verrucae: a follow-up study. AB - In this report, the authors will supplement research performed in 1989 by Sollitto, Napoli, and Gazivoda on the use of intralesional bleomycin for the treatment of verrucae. With various alterations of technique, their original success rate of 32.2% was improved to 65.4%. Thus, bleomycin is a very effective first line treatment for multiple verrucae, particularly of the mosaic variety. An association is also made between patients with pedal hyperhidrosis and the occurrence of multiple verrucae, notably lesions that prove recalcitrant to initial therapy. PMID- 8722888 TI - Radiologic identification of Ewing's sarcoma. AB - The authors discuss Ewing's sarcoma, a rare pedal neoplasm. They concentrate on radiologic interpretation of this entity. Case studies are illustrated. PMID- 8722889 TI - The Valenti procedure for hallux limitus/rigidus. PMID- 8722890 TI - The Valenti procedure for hallux limitus/rigidus. PMID- 8722891 TI - Methicillin-resistant coagulase-negative Staphylococcal osteomyelitis and its relationship to broad-spectrum oral antibiosis in a predominantly diabetic population. PMID- 8722892 TI - The medial instep plantar fasciotomy. PMID- 8722893 TI - A retrospective comparison of endoscopic plantar fasciotomy to open plantar fasciotomy with heel spur resection for chronic plantar fasciitis/heel spur syndrome. PMID- 8722894 TI - Giant cell tumor of tendon sheath. PMID- 8722895 TI - Buried Kirschner wire fixation in digital fusion. PMID- 8722896 TI - Comparative study of oral and written picture description in patients with Alzheimer's disease. AB - Oral and written picture descriptions were compared in 22 patients with Alzheimer's disease (AD) and 24 healthy elderly subjects. AD patients had a significant reduction of all word categories, which, similarly to controls, was more pronounced in written than in oral texts. They also reported fewer information units than controls, but without task difference. At the syntactic level, written descriptions of AD subjects were characterized by a diminution of subordinate clauses and a reduction of functors. More grammatical errors were present in written descriptions by AD and control subjects. AD and control groups produced an equivalent number of semantic errors in both tasks. However, in oral description, AD patients had more word-finding difficulties. In sum, AD descriptions were always shorter and less informative than control texts. Additionally, written descriptions of AD patients appeared shorter and more syntactically simplified than, but as informative as oral descriptions. Whereas no phonemic paraphasias were observed in either group, AD patients produced many more graphemic paragraphias than controls produced. Furthermore, written descriptions had more irrelevant semantic intrusions. Thus, as compared to oral descriptions, written texts appeared to be a more reliable test of semantic and linguistics difficulties in AD. PMID- 8722898 TI - Theory of mind and pragmatic understanding following right hemisphere damage. AB - It has been maintained that 3-year-olds' difficulties in correctly predicting the undesired outcome of false beliefs reflects difficulties in interpreting the implications of conversations rather than a conceptual limitation in their theory of mind. As the right hemisphere has been seen to be responsible for the interpretation of the pragmatic aspects of communication, right-hemisphere damaged (RHD) and left-hemisphere-damaged (LHD) adult patients in our study were compared on their ability to correctly draw inferences in false belief tasks. The RHD but not the LHD patients were found to have difficulties similar to those of young children in understanding the conversational implications test questions. Most reported that a central story character would look for a pet in the place where it was really located instead of where the character believed it was located. However, when then asked in a control question where the pet really was, the RHD patients often switched their answer to the test question and referred to the believed location. Removal of the need to infer the questioner's meaning enabled both RHD and LHD subjects to make correct false belief predictions. The results are discussed in terms of the effects of brain damage on spatial memory and the pragmatic demands of theory of mind tasks. PMID- 8722897 TI - A positron emission tomography study of binaurally and dichotically presented stimuli: effects of level of language and directed attention. AB - Regional cerebral blood flow (rCBF) was measured using positron emission tomography with oxygen- 15 labeled water as 10 normal subjects listened to three types of auditory stimuli (environmental sounds, meaningless speech, and words) presented binaurally or dichotically. Binaurally presented environmental sounds and words caused similar bilateral rCBF increases in left and right superior temporal gyri. Dichotically presented stimuli (subjects attended to left or right ears) caused asymmetric activation in the temporal lobes, resulting from increased rCBF in temporal lobe regions contralateral to the attended ear and decreased rCBF in the opposite hemisphere. The results indicate that auditorily presented language and non-language stimuli activate similar temporal regions, that dichotic stimulation dramatically changes rCBF in temporal lobes, and that the change is due both to attentional mechanisms and to hemispheric specialization. PMID- 8722899 TI - Differences in susceptibility to the "blending illusion" among Native Hebrew and English speakers. AB - This study was designed to explore whether native Hebrew and English speakers differ in their susceptibility to the blend illusion and in their lip-reading of words and cliche sentences. Four lip-reading tests, designed to assess recognition of syllables, words, and overlearned sentences, were administered to two groups of subjects who were matched for age, sex, and educational level. The English-speaking group showed significantly higher susceptibility to the blending illusion. The source of the effect was in the enhanced sensitivity to the auditory input in males along with reduced number of auditory biases in females. No sex differences in lip-reading of any item of the lip-reading battery was obtained in native Hebrew speakers. Subjects in both groups performed at the same level in recognizing the words and overlearned sentences. The result is attributed to the shorter Voice Onset Time values of Hebrew when native listeners would hear acoustic messages prior to discernable lip movements. PMID- 8722900 TI - Knowledge of scripts reflected in discourse of aphasics and right-brain-damaged patients. AB - Script knowledge was examined in discourse produced by 15 mildly moderately impaired aphasics, 15 right-hemisphere-damaged (RHD) subjects. 15 neurological patients without central nervous system damage, and 30 normal controls. Experimental task were designed to elicit knowledge of particular scripts, various scripts from a category, metascripts, and plans. The results showed that the brain-damaged patients were able to produce information belonging to a given semantic category of scripts. Disorders demonstrated by both aphasics and RHD patients involved different discourse processes. Aphasics' impairment was mainly connected with discourse macroprocessing. Disturbances in the RHD subjects were found in text structures, contents, and macrostructure. The problem of the probable mechanisms of the observed discourse disturbances were discussed. PMID- 8722901 TI - Clinical insights into pragmatic theory: frontal lobe deficits and sarcasm. AB - The validity of psycholinguistic theories of sarcasm was explored by examining subjects with mainly frontal lobe (FL) damage and concomitant concreteness of thought. The majority of FL subjects could interpret consistent verbal exchanges but not literally contradictory (sarcastic) verbal exchanges which implied that the literal meaning of a sarcastic comment needs to be rejected in order for the inference to be detected. Subsidiary analyses confirmed that failure on sarcasm tasks was associated with poor conceptual skills. Ability to process attitude was not associated with success at recognizing sarcasm. This suggested that attitude is not pivotal to the detection of sarcastic inference. PMID- 8722903 TI - Hemispheric independence in word recognition: evidence from unilateral and bilateral presentations. AB - We compared behavioral laterality effects in a lexical decision task using cued unilateral or bilateral presentations of different stimuli to normal subjects. The goals were to determine the effects of lexical variables on word recognition in each hemisphere under conditions of maximal independence of information processing in the two hemispheres and to assess the degree of residual interhemispheric effects that can still exist then. Bilateral presentations increased hemispheric independence in word recognition, indexed by a significant interaction of response hand with target visual field. Bilateral presentations also selectively impaired word decisions, suggesting that word processing benefits from interhemispheric interactions, whereas nonword processing is done independently in each hemisphere. Indeed, there was a significant congruity effect for word targets only, whereby the wordness of the unattended stimulus affected the speed of processing of attended word targets. Word frequency and regularity affected both hemispheres equally, arguing against the hemispheric interpretation on the dual route model of word recognition. Length affected the processing of nonwords more than words and in the left visual field more than in the right visual field. Taken together, the data support the conclusion that each normal hemisphere can control word recognition independently of the other. PMID- 8722902 TI - Picture confrontation oral naming: performance differences between aphasics and normals. AB - Oral confrontation naming was compared in 108 normal subjects controlled for education, age, and gender and in 18 aphasic patients for the same set of 115 pictures. Demographic variables influenced both normals' and aphasics' performance. However, the nature of aphasics' misnamings on the one hand and the differential effects of characteristics of pictures and words on normals' and aphasics' responses on the other indicated specific deficits in patients. The classical hypothesis that aphasics' misnamings and the production of word associations by normals should rely on similar mechanisms (Rinnert & Whitaker, 1973) is questioned. Nondominant responses observed in normals accounted for a larger proportion of verbal errors than associates to target words. PMID- 8722904 TI - Use of randomly amplified polymorphic DNA markers for the genetic analysis of relatedness and diversity in chickens and turkeys. AB - A study involving the use of random amplified polymorphic DNA (RAPD) was conducted to evaluate genetic polymorphism and relatedness within and among four chicken breeds: Araucona, Rhode Island Red, White Leghorn, and White Plymouth Rock, and two turkey populations, a long-term randombred and a commercial strain. A total of 60 random primers were used in the RAPD analyses. Forty-two of the 60 primers tested amplified patterns with at least one polymorphic fragment in one or more of the populations. Six of these 42 primers amplified polymorphic fragments in each of the six strains with a within- and between-population average band-sharing frequency of less than one but above zero (P < 0.05). Differences among the six primers for genetic distance (D) among populations were significant (P < 0.05). A consensus dendogram was therefore developed to show the phylogenetic relationships among the populations. As expected, estimates of D between populations were lowest within species and highest between species. The results provide evidence of the applicability of RAPD to determining genetic relatedness within and among different poultry populations and in developing reproducible markers useful in evaluating individual variation in chickens and turkeys. PMID- 8722905 TI - A method for separating sex-linked imperfect albino (S*ALS) and nonalbino embryos before hatch. AB - The feasibility of using the sex-linked gene for imperfect albinism (S*ALS) to sex chicks during incubation by candling was studied. With this technique, the dark eye of nonalbino embryos can be positively identified. Two trials were performed. In a first trial, 66.5 and 89.5% of the 254 nonalbino and 210 albino chicks produced in four hatches were correctly identified by candling at 7 d of incubation. Of 191 eggs predicted to be nonalbinos, 22 were albinos, resulting in an overall accuracy of 88.5% for identification of nonalbino embryos. In a second trial, the accuracy of the technique from 7 to 10 d of incubation was evaluated. Increased age resulted in a tendency for lower accuracy, but candling at 8, 9, or 10 d of incubation allowed identification of a greater (P < 0.05) proportion of the nonalbino population than at 7 d of incubation. Candling at 8 d of incubation allowed identification of nonalbinos and albinos with an accuracy of 81.3 and 84.9%, respectively, suggesting that the ALS gene could be used to sex chicks during incubation when used in a sex-linked cross. This technique may prove advantageous to the laying industry because of savings of incubator and hatcher space. The males of commercial layer lines are normally killed at hatch. Reducing the number that hatch by eliminating them before 10 d of incubation could diminish animal welfare concerns. PMID- 8722906 TI - The use of intermittent lighting in broiler raising. 1. Effect on broiler performance and efficiency of nitrogen retention. AB - The purpose of the present experiments was to compare the effects of the intermittent lighting schedule (IL) consisting of repeated cycles of 1 h light (L):3 h darkness (D) with a nearly continuous lighting schedule 23L:1D (CL) on the performance of floor-reared male and female broiler chickens. In addition, N balance trials were performed on caged broilers in order to assess their efficiency of dietary N retention. Changing from CL to IL at Day 7 was followed by a temporary reduction in feed intake and BW gain. However, IL males exhibited compensatory growth during the finisher period in such a way that at Day 41 (market age), a similar BW as that of their CL counterparts was attained. In contrast, IL females did not completely attain a BW equal to that of CL females by 41 d of age. Imposing IL reduced feed intake and improved feed conversion. Abdominal fat pad contents of broilers reared under IL were significantly smaller than those of CL broilers. From the N balance trials, it can be inferred that broilers reared under IL have an improved efficiency of dietary N retention, especially during the period of increased growth velocity. Imposing IL at Day 7 is associated with a temporary reduction in BW gain, followed by complete growth compensation of males by Day 41. Female broilers did not restore BW by this age. As a consequence of the altered growth trajectory, the manifestation of the fat growth wave is shifted to an older age together with an enhanced efficiency of N retention. PMID- 8722908 TI - An application of the augmented randomized complete block design to poultry research. AB - Avian health researchers are often faced with the need to evaluate large numbers of potential disease agents, but limited facilities and the desire to minimize animal usage and costs prohibit large, all-inclusive experiments. Plant breeders encounter a similar problem with variety screening experiments in which a large number of varieties must be evaluated, but the supply of seed may be limited. In that situation, the use of augmented experimental designs has proven effective for obtaining the replication needed for valid statistical analysis. Those designs rely on the use of multiple trials consisting of different sets of plant varieties; standard varieties are included as controls or checks in every trial to provide an estimate of experimental error. Although the comparisons among varieties are not all equally precise and the estimate of experimental error may be biased, use of augmented designs can reduce the time and number of experimental units needed to isolate the most promising new varieties for further in-depth study. Augmented designs can similarly be applied to screening experiments in poultry research. This paper describes the augmented randomized complete block design and its application in an avian health setting. Data collected in an exploratory study of infectious enteritis in chickens illustrate details of the statistical analysis. Suggestions for implementing this analysis using the SAS statistical analysis package are also given. PMID- 8722907 TI - The use of intermittent lighting in broiler raising. 2. Effects on the somatotrophic and thyroid axes and on plasma testosterone levels. AB - Male and female broiler chicks were raised separately in nearly continuous lighting [23 h light (L):1 h dark (D), CL] and consumed feed ad libitum. At 7 d of age, the intermittent lighting schedule (1L:3D, IL) was imposed on half of the chicks, whereas the other chicks remained under CL. In addition to performance characteristics, several parameters of the somatotrophic and thyrotrophic axes were studied together with plasma concentrations of testosterone. Males had a higher growth rate than females regardless of the imposed lighting schedule and this pronounced growth difference is reflected by higher plasma concentrations of growth hormone (GH), and a better GH receptor occupancy. Differences in growth rate between sexes could not be attributed to differences in circulating 3,3',5 triiodothyronine (T3) levels or to hepatic deiodination activities. However, from 3 wk of age onwards, males had significantly higher plasma testosterone levels than females. Plasma GH and T3 levels decreased whereas plasma insulin-like growth factor-I and thyroxine levels increased with age in all experimental groups. The age-related decline in plasma GH levels were less pronounced for males than for females. No major changes in other hormonal parameters or deiodination activities could be observed as a result of imposing IL, except for the higher plasma GH levels of IL chickens, and for plasma testosterone concentrations in IL males at Day 41, which were twice the levels found in their CL counterparts. These results therefore suggest that the somatotrophic axis as well as circulating testosterone levels mediate the sex-related differences in growth rate and the compensatory growth as present in males. PMID- 8722909 TI - Biodegradability and microbial activities during composting of poultry litter. AB - Poultry litter is composted to reduce odor and pathogens and to improve its quality as a soil amendment. Organic material, e.g., sawdust, is added to increase the C:N ratio to achieve optimum degradation of organic C and retention of N through microbial biomass formation. However, the relative biodegradabilities of the organic material in poultry litter and the amendment are usually not known. Furthermore, it is assumed that as microorganisms metabolize organic compounds and produce CO2, they increase in biomass and, therefore, retain N. In this study, bench-scale compost reactors were used to determine the relative contributions of poultry litter and of the amendment (sawdust) to the biodegradability of a compost mix. Approximately 29% of the volatiles lost from the poultry litter mix came from the sawdust. Fiber analyses revealed that only a small portion of cellulose was degraded. Although microbial subpopulations able to degrade selected macromolecules were present at varying levels, the overall level of microorganisms did not change markedly. Populations capable of degrading bacterial cell walls were present throughout the composting period, and microbiological assays indicated that inorganic nutrients were available to support limited microbial growth. These results suggest that N compounds and inorganic nutrients are recycled, rather than fixed during composting. PMID- 8722910 TI - Effects of supplemental phytase and phosphorus on histological and other tibial bone characteristics and performances of broilers fed semi-purified diets. AB - Two trials with day-old chicks were conducted to investigate the effects of supplemental phytase (Natuphos) on histological, mechanical, and chemical properties of tibia, and performances of broilers fed semi-purified diets containing soybean meal as the only organic P source [0.11% nonphytate P (nP)]. Dietary treatments in Trial 1 were: 1) 0.20% nP, 2) Diet 1 + 800 U of phytase/kg of diet, 3) 0.27% nP, 4) Diet 3 + 600 U of phytase, 5) 0.34% nP, 6) Diet 5 + 400 U of phytase. Supplemental phytase and inorganic P increased tibial length (P < 0.01), shear force (P < 0.001), shear stress (P < 0.05), ash content (P < 0.001), and BW gain and feed intake (P < 0.001) during Trial 1. The hypertrophic zone width at the proximal end of the tibia was decreased (P < 0.05), and the tibial width (P < 0.05) of the long axis of the tibia was increased by the phytase and P supplementation. Supplemental phytase enlarged the cartilaginous and proliferative zones of the tibial proximal end (P < 0.05), and an increase in nP levels produced similar effects. Supplementation of phytase and P also tended to improve the orderliness of development and arrangement of cartilage and bone cells. Dietary treatments in Trial 2 were: 1) 0.27% nP, 2) Diet 1 + 350 U of phytase, 3) Diet 1 + 1,050 U of phytase, 4) 0.45% nP, 5) 0.54% nP, 6) Diet 5 + 1,050 U of phytase. Broilers fed diets containing relatively high levels of nP and phytase supplementation in Trial 2 gave results similar to those observed in Trial 1. Marked improvements (P < 0.05) in the ash content, shear force, shear stress, length of tibia, BW gain, and feed intake, and reduced hypertrophic zone width were achieved for broilers fed the P-deficient diet supplemented with phytase. Also, supplemental phytase tended to increase the width of cartilaginous and proliferative zones, to increase trabecular bone density, and to improve the orderliness of development and mineralization of cartilage and bone cells. In summary, supplementing a low-nP diet with inorganic P or phytase resulted in similar beneficial effects on bone development. PMID- 8722911 TI - Age-related changes of protein turnover in specific tissues of the chick. AB - The effect of age on muscle and liver protein turnover was analyzed in growing broilers. Protein turnover was compared in the Pectoralis major (PM), the Anterior Latissimus dorsi (ALD), and the Sartorius (SART) muscles, as well as in the liver in broilers at three ages (2, 3, and 4 wk). Protein fractional synthesis rates (FSR, in percentage per day) were measured by a flooding dose of L-[4-3H] Phe. Protein fractional breakdown rates (FBR, in percentage per day) were estimated as the difference between FSR and the gain rates of tissue protein. When expressed in absolute rates (grams per day), tissue protein deposition increased during chick growth (approximately 1.7-fold, whatever the tissue). This phenomenon was accompanied by increased protein synthesis (ASR) and proteolysis (ABR) in the PM and in the ALD muscles as well as in the liver. However, ASR and ABR did not seem to be significantly modified in SART. The FSR in skeletal muscles significantly decreased with age. This developmental decline was associated with decreases in RNA content and RNA: protein ratio (Cs, capacity for protein synthesis). In contrast to muscle, we found no significant variation in liver FSR between 2 and 4 wk of age. In addition, liver Cs and translational efficiencies were not decreased. Finally, in muscle as in liver, FBR did not show any clear age-related pattern. PMID- 8722912 TI - Interrelationship between the effects of dietary factors and feed withdrawal on the content and composition of liver fat in broiler chicks. AB - Four experiments were carried out to evaluate the effects of the composition and pelleting of diets fed to broiler chicks up to marketing age on changes in their liver size and the content and composition of liver fat due to feed withdrawal (FW) during 0, 10, and 24 h. Birds not exposed to FW that were fed diets with high energy to protein ratio (E:P) and diets in pelleted from-diets that increase fattening-had significantly (P < 0.05) higher liver fat content than those fed diets with the recommended E:P and in mash form, respectively. Those fed pellets also had higher liver weight. Dietary energy level did not affect these variables. Feed withdrawal for 10 or 24 h decreased, at times significantly, liver weight and its fat content, irrespective of the diets fed previously. The values observed after FW were not affected by the dietary factors evaluated. The composition of liver fatty acid in chicks not exposed to FW was markedly affected by increases in dietary energy (soybean oil) level and E:P. The main effects of the former were increased levels of stearic, linoleic, and arachidonic acids and a decreased level of oleic acid; those of the latter were an increased level of oleic and decreased levels of stearic, linoleic, and arachidonic acids. Irrespective of the diets used, FW increased the concentrations of stearic, linoleic, and arachidonic acids, and decreased those of palmitic and oleic acids. This effect on arachidonic acid was consistently significant, whereas the effects on the other fatty acids were significant in two out of three experiments. Due to these effects of FW, part of the differences in the composition of liver fatty acids caused by dietary factors observed in fed chicks, completely disappeared after FW. The length of FW (10 vs 24 h) did not affect liver size or fat content, but the concentrations of oleic and arachidonic acids in liver fat were significantly higher in birds exposed to 24 h of FW. It was concluded that the composition and form of the diets markedly affect liver weight and the content and composition of its fat in birds not exposed to FW. The effects of the dietary factors on liver size and its fat content completely disappear after 24 h of FW. PMID- 8722913 TI - Mapping functional chicken genes: an alternative approach. AB - Functional genes were selected for linkage analysis mapping using the East Lansing (EL) reference population ?[Jungle Fowl (JF) x White Leghorn (WL)] x WL?. The approach used was based on the identification of DNA sequence polymorphisms in the introns of those genes found in JF and WL. Deoxyribonucleic acid sequence analysis revealed single base substitutions in introns of six Type I marker genes: adenylate kinase 1 (AK1), aldolase B (ALDOB), a lysosomal membrane protein gene (LAMP1), vitellogenin 2 (VTG2), apolipoprotein A1 (APOA1), and creatine kinase B (CKB). Transitions or transversions were found in introns of AK1, ALDOB, LAMP1, VTG2, APOA1, and CKB. A transversion in the intron of the JF allele of AK1 generated a unique BspHI cleavage site. The design of polymerase chain reaction (PCR) primers based on the site of base substitution led to the specific amplification of the JF allele in the remaining five genes. A size polymorphism in the PCR production derived from iron response element binding protein (IREBP) distinguished the JF from the WL allele. Linkage analysis of the EL reference population revealed that these candidate genes were located in the following EL linkage groups (E) or chromosomes (Chrom) of the chicken genome: AK1 (E41), VTG2 (E43), APOA1 (E49), CKB (E07), LAMP1 (E01), ALDOB (Chrom Z), and IREBP (Chrom Z). Provided that a base substitution can be found in the parents of the reference population, this PCR-based approach can be used to map any cloned candidate gene. This approach will lead to further information on synteny of the chicken genome with cognate genes of mammalian species. PMID- 8722915 TI - Relationship of eggshell ultrastructure and shell strength to the soundness of shell eggs. AB - Factors affecting the soundness of shell eggs are of primary concern to egg processors due to substantial financial losses from cracked and leaker eggs. Ultrastructural analyses were used to examine the palisade layer width and mammillary knob layer thickness of sound, cracked, and leaker eggshells. Subjective observations were also made. There was no significant difference (P > 0.05) in the width of the palisade layer or the mammillary knob layer among sound, cracked, and leaker eggshells. The eggshell strengths of sound, cracked, and leaker eggs were evaluated using puncture force and shell thickness measurements. Sound eggshells were found to have a significantly (P < 0.05) higher mean puncture force (35.3 N) than cracked (30.4 N) and leaker (28.4 N) eggshells. The mean puncture force values for cracked and leaker eggshells were not significantly different from each other (P > 0.05). Regression and correlation analyses indicated a significant correlation (P < 0.01; r = 0.61) between palisade layer width and puncture force. The mean shell thickness values for cracked (0.36 mm) and leaker (0.35 mm) eggs were significantly lower (P < 0.05) than those of sound eggs (0.38 mm), although cracked and leaker eggshell thickness did not differ significantly (P > 0.05). The correlation coefficient between puncture force and shell thickness was significant (P < 0.01; r = 0.56), indicating a possible relationship. PMID- 8722914 TI - Effects of an in ovo injection of an anti-androgen on embryonic and posthatching growth of broiler chicks. AB - Experiments were conducted to determine whether in ovo injection of Flutamide, a non-steroidal anti-androgen, would affect posthatching growth of broiler chickens. Before doing these studies it was necessary to develop a bioassay to demonstrate that Flutamide had anti-androgenic actions when injected in ovo. In ovo injection of 0.06 to 1.74 mumol of testosterone propionate (TP) per egg on the 8th d of incubation caused a dose-related decrease in weight of the bursa of Fabricius of newly hatched chicks. In subsequent experiments, 1.74 mumol of Flutamide significantly decreased the bursal regression produced by 0.58 mumol of TP when both substances were injected on Day 8 of incubation and bursal weights were taken on the day of hatch. Having established that Flutamide could block the effects of high doses of exogenous TP, it was reasoned that it could block the action of much lower endogenous levels. In two of the three experiments, injection of Flutamide on the 8th d of incubation resulted in a dose-related decrease in 21- and 49-d BW of male broiler chickens. The material had no effect on BW of males in one experiment and had no effect on BW of females at any age in any of the experiments. The data suggest that interference with the actions of endogenous androgens during embryonic life suppresses the posthatching growth of male broilers. PMID- 8722916 TI - The effects of alternative electrical and mechanical stunning methods on hemorrhaging and meat quality of broiler breast and thigh muscles. AB - Five trials were conducted to determine the effects of various alternative stunning and restraining methods on the quality of chicken broiler meat. The stunning methods used were electrical whole-body and head-only stunning with different voltages (25 vs 100 V) and frequencies (50 vs 200 Hz), and mechanical captive bolt stunning. Before being stunned, the broilers were restrained either by shackling them by the feet, restraining their heads while they were standing, or placing them in a cone. Convulsions during and after stunning were subjectively scored. At 1 d post-mortem, the pH, color, and the intensity and number of hemorrhages, of breast and thigh muscles were measured. Convulsions were, in order of increasing intensity, observed after whole-body stunning using 50 Hz, whole-body stunning using 200 Hz, head-only stunning, and captive bolt stunning. No differences in ultimate pH were measured. A consistent difference in color was observed between whole-body and head-only stunning. Head-only stunning resulted in darker and more red (P < or = 0.01) breast muscles. Head-only and captive bolt stunning resulted in hemorrhages that were located more distal in the breast muscle than those observed after whole-body stunning. Captive bolt stunning (in a cone) resulted in fewer (P < or = 0.01) hemorrhages in breast and thigh muscles than whole-body stunning (100 V, 50 Hz, shackled), and in a similar level of hemorrhaging as electrical head-only stunning (25 V, 200 Hz, in a cone). PMID- 8722917 TI - The effects of stunning amperage and deboning time on early rigor development and breast meat quality of broilers. AB - Two trials were conducted to determine the effects of stunning amperage on the early rigor development and quality of broiler breast meat. In each trial, 108 broilers were killed either without stunning or following constant amperage stunning at 50 or 125 mA. The breast muscles (Pectoralis major) were removed from the carcasses at 0.2, 1, 2, 4, 6, 8, 10, 12, or 24 h post-mortem. Breast muscle pH and R-value (inosine to adenosine nucleotide ratio) were determined immediately upon deboning on one side of each carcass. After aging the deboned breast fillets for 48 h, cook loss and Allo-Kramer shear values were determined on the opposite side. Stunning at 50 or 125 mA resulted in significantly higher muscle pH values up to 6 h post-mortem, after which there were no further significant differences. Stunning at 50 and 125 mA resulted in significantly lower R-values up to 6 h post-mortem. Stunning had no effect on cook loss. Stunning at 125 mA resulted in significantly tougher meat up to 10 h post-mortem than meat from unstunned birds or birds stunned at 50 mA. These results indicate that the delayed onset of rigor, as measured by muscle pH and R-value, noted in electrically stunned birds lasts for approximately 4 to 6 h, after which differences due to stunning disappear. PMID- 8722918 TI - Effect of stunning time and polyphosphates on quality of cooked chicken breast meat. AB - The objective of this research was to determine effects of stunning duration and polyphosphates on color, pH, and texture of chicken breast meat. Sixty broiler chickens were electrically stunned for 2 to 10 s in 2-s intervals and then slaughtered. Left and right Pectoralis major muscles were excised 1 h post mortem. The left muscles (treated) were treated with a sodium tripolyphosphate salt solution, and the right muscles (controls) were treated with a salt solution alone. All muscles were cooked to an endpoint temperature of 72 C in a 95 C water bath. Measurements of muscle pH were taken on the meat 1 h post-mortem, after treating, and after cooking. Color values and Warner-Bratzler shear values were evaluated on each cooked meat sample. As stunning time increased, pH increased at 1 h post-mortem but treating with either of the solutions eliminated this trend. Muscle pH also increased upon cooking except in the polyphosphate-treated muscles, because the latter muscles achieved maximum pH prior to cooking. Stunning had no apparent effect on color values of the cooked meat; however, polyphosphate treatment resulted in meat that was darker and less red than controls. Longer stunning times resulted in greater shear values but polyphosphates did not affect Warner-Bratzler results. Taken together, these data suggest that processing variables that affect muscle pH also affect quality attributes. PMID- 8722919 TI - Dispelling the myths of publishing. PMID- 8722920 TI - Patients' perceptions of environmental factors that disturb sleep after cardiac surgery. AB - BACKGROUND: Effective management of sleep disturbances after cardiac surgery requires insight into patients perceptions of which factors disturb sleep in the intensive care unit and after transfer from the intensive care unit. OBJECTIVE: To describe patients perceptions of environmental factors that disturbed sleep after cardiac surgery. SAMPLE: A convenience sample of 102 patients was surveyed in a 300-bed, acute-care teaching hospital. METHOD: Patients were interviewed several days before discharge from the hospital and rated the extent to which specific factors disturbed their sleep while in the intensive care unit and after transfer. RESULTS: The group mean for the extent of disturbance scores, averaged across all 35 environmental factors studied, was low during and after stay in the intensive care unit. However, selected items such as pain, and inability to get comfortable and perform a familiar nighttime routine, were moderately disturbing to many patients across phases of recovery. Patients varied widely in the number of factors, ranging from 0 to 33 factors, that disturbed sleep at least to some extent. Patients added factors that were not included in the original monitoring instrument. CONCLUSIONS: Patients attribute disturbed sleep to specific factors after cardiac surgery. The number of factors that disturb sleep varies among patients. Nurses can modify many of the factors that disturb sleep to promote an environment that will facilitate improved sleep, thereby enhancing the acute phase of recovery from cardiac surgery. PMID- 8722921 TI - Individual factors that influence sleep after cardiac surgery. AB - BACKGROUND: Up to 50% of patients who undergo cardiac surgery report problems with sleep after cardiac surgery. Knowledge about which individual factors are associated with sleep problems after cardiac surgery would help nurses identify patients who are at greater risk for sleep problems during hospitalization. OBJECTIVE: To compare patients perceptions of sleep before and during hospitalization for cardiac surgery and identify and analyze individual factors in relation to patients perceptions of sleep. METHOD: A sample of 102 patients who underwent elective or emergent cardiac surgery were studied at a 300-bed teaching hospital in the northwestern United States. A few days before their anticipated discharge from the hospital, consenting patients completed questions about their sleep before hospitalization and the night before their interview. RESULTS: Patients reported that they slept fewer hours in the hospital than at home. No differences were found in patients perceived depth and sufficiency of sleep, or refreshment before and after surgery. Patients who slept poorly at home did not report any worse sleep after surgery than patients who slept well at home. Women's perceptions about the sufficiency, refreshment, and quality of sleep were consistent before and after surgery, but no relationships were found among men's ratings. The length of sleep at home was positively related to the length of sleep after surgery in older patients. CONCLUSIONS: Sleep length is related to patients perceptions of sleep after cardiac surgery. Gender and age are also related to qualitative aspects of sleep before and after surgery and can be instrumental in an individualized assessment of sleep patterns anticipated after cardiac surgery. PMID- 8722923 TI - ST segment monitoring with a derived 12-lead electrocardiogram is superior to routine cardiac care unit monitoring. AB - BACKGROUND: Prior studies have shown that a derived 12-lead electrocardiogram with a simple electrode configuration is comparable with the standard electrocardiogram for arrhythmia analysis. METHODS: A prospective, comparative, within subjects design was used to compare the value of the derived 12-lead electrocardiogram with that of routine monitoring of leads V1 and II for detection of transient myocardial ischemia in 250 patients treated for unstable angina or myocardial infarction. RESULTS: During 11,532 hours of derived 12-lead ST segment monitoring, 55 (22%) of 250 patients had 176 episodes of ischemia. Of the 55 patients with ischemia, 75% reported no chest pain and 64% had no ischemic ST changes with routine monitoring leads. All five patients who developed angiographically confirmed abrupt reocclusion after percutaneous transluminal coronary angioplasty had ischemic ST changes with the derived electrocardiogram (sensitivity, 100%), compared with only two patients with routine monitoring (sensitivity, 40%). Serious complications occurred in 17% of angina patients with ischemic events compared to 3% of those without ischemia. Length of stay in the cardiac care unit was twice as long in angina patients who had ischemic events. In patients with acute myocardial infarction, ischemic events were not associated with a more complicated hospital course; however, length of stay in the cardiac care unit was longer in patients with recurrent ischemia. CONCLUSIONS: The findings show that derived 12-lead ST monitoring is superior to routine monitoring of leads V1 and II for detecting transient myocardial ischemia. ST monitoring of the derived 12-lead electrocardiogram may identify high-risk patients with unstable angina and provide prognostic information that would not be otherwise available from the usual clinical measures. PMID- 8722922 TI - Endotracheal suctioning: ventilator vs manual delivery of hyperoxygenation breaths. AB - BACKGROUND: Despite a large number of studies on endotracheal suctioning, there is little data on the impact of clinically practical hyperoxygenation techniques on physiologic parameters in critically ill patients. OBJECTIVE: To compare the manual and mechanical delivery of hyperoxygenation before and after endotracheal suctioning using methods commonly employed in clinical practice. METHODS: A quasi experimental design was used, with twenty-nine ventilated patients with a lung injury index of 1.54 (mild-moderate lung injury). Three breaths were given before and after each of two suction catheter passes using both the manual resuscitation bag and the ventilator. Arterial pressure, capillary oxygen saturation, heart rate, and cardiac rhythm were monitored for 1 minute prior to the intervention to obtain a baseline, continuously throughout the procedure, and for 3 minutes afterward. Arterial blood gases were collected immediately prior to the suctioning intervention, immediately after, and at 30, 60, 120, and 180 seconds after the intervention. Data were analyzed with repeated measures analysis of variance. RESULTS: Arterial oxygen partial pressures were significantly higher using the ventilator method. Peak inspiratory pressures during hyperoxygenation were significantly higher with the manual resuscitation bag method. Significant increases were observed in mean arterial pressure during and after suctioning, with both delivery methods, with no difference between methods. Maximal increases in arterial oxygen partial pressure and arterial oxygen saturation occurred 30 seconds after hyperoxygenation, falling to baseline values at 3 minutes for both methods. CONCLUSION: Using techniques currently employed in clinical practice, these findings support the use of the patient's ventilator for hyperoxygenation during suctioning. PMID- 8722924 TI - Technology assessment in critical care: understanding statistical analyses used to assess agreement between methods of clinical measurement. AB - Many new measurement methods that employ various technologies to measure physiological parameters have been introduced into the field of critical care. Clinical assessment of these new methods occurs through the conduct of method comparison studies in which the level of agreement between a new measurement method and a clinical standard method is determined. Clinicians and researchers are often faced with the complicated task of analyzing and interpreting the results of method-comparison studies. Use of correlation and linear regression techniques has been prevalent in method-comparison studies but has proven inappropriate and inadequate in determining how well methods compare. The purposes of this article are to briefly review the terms of accuracy, agreement, and the precision in context with method-comparison studies, and discuss inappropriate and appropriate statistical analyses and their interpretation. Appropriate data analysis of method-comparison studies will aid in determining not only whether new monitoring methods can be interchanged or used in place of existing methods, but whether new methods warrant further research of their effect on patient outcomes. PMID- 8722925 TI - A guideline for competency of the critical care nurse. AB - BACKGROUND: To maintain a high standard of quality nursing care the individual critical care nurse s competencies should be determined objectively. Few international guidelines describe the qualities required by critical care nurses to function effectively in a critical care unit. These guidelines often focus only on the education and training of critical care nurses. OBJECTIVE: To formulate and describe guidelines for the competency requirements of registered critical care nurses. METHOD: A focus group interview, a qualitative research method, was conducted as an open conversation in which each participant could ask questions, comment, or respond to comments. Interaction among the respondents was encouraged to stimulate in-depth discussion. The study was conducted within the framework of South African critical care nursing. RESULTS: The four main categories that were identified are as follows: professional competence, cognitive competence, interpersonal skills, and critical care patterns of interaction. These are described in detail along with a formulated guideline for critical care nurse competency requirements. CONCLUSION: This study describes an attempt to formulate objective guidelines for critical care nurses competency requirements. PMID- 8722927 TI - Pleural empyema. PMID- 8722926 TI - Hypertension, stroke, and noncompliance: an avoidable triad. PMID- 8722928 TI - Photo quiz. Sporotrichosis. PMID- 8722929 TI - General principles of specimen collection and transport. AB - In this issue of Clinical Infectious Diseases, we present the first article in a series entitled "Diagnostic Microbiology Updates." Although clinical microbiology is included in the curricula of virtually all infectious disease fellowships, the degree of emphasis on this subject varies considerably. Infectious disease physicians--even those who have direct responsibilities or consulting responsibilities for the microbiology laboratories of the institutions in which they practice--may be hard pressed to keep up with the rapidly changing content of the primary literature in clinical microbiology. The purpose of this series, therefore, is at least in part to fill this void and to provide concise updates for clinicians. The first article, written by Dr. Michael L. Wilson, reviews current concepts in specimen collection and transport. A key issue for all clinicians (which is not always sufficiently emphasized) is the quality of the specimen submitted to the laboratory. It is an axiom that if specimens of poor quality are submitted, the results generated by the laboratory will have little or no clinical utility. Dr. Wilson's article describes some of the methods available to assure that only specimens of good quality, i.e., those most likely to be useful clinically, are processed in the microbiology laboratory. Future articles will address specific types of specimens, groups of pathogens, and diagnostic techniques, including molecular methods. We hope this series will be informative and valuable to the readers of Clinical Infectious Diseases, and we look forward to your comments. PMID- 8722930 TI - Community respiratory virus infections among hospitalized adult bone marrow transplant recipients. AB - From 1 November 1992 through 1 May 1993 and from 1 November 1993 through 1 May 1994, we conducted a prospective surveillance study at the University of Texas M.D. Anderson Cancer Center (Houston) to evaluate the role of community respiratory virus infections in hospitalized adult bone marrow transplant (BMT) recipients, Respiratory secretions were obtained from all adult BMT recipients with acute respiratory illnesses. During these two winters, a community respiratory virus was isolated from 37 (36%) of 102 patients and 30 (26%) of 115 patients, respectively. Approximately half (49%) of these infections were due to respiratory syncytial virus (RSV); the remainder were due to influenza virus (18%), picornaviruses (18%), parainfluenza virus (9%), or adenovirus (6%). Fifty eight percent of these infections were complicated by pneumonia, with an associated mortality of 51%. The pneumonias that complicated RSV infection were almost exclusively viral in origin and were associated with a mortality of 100% if not treated promptly with antiviral agents. In contrast, many of the pneumonias that complicated the other viral infections, such as influenza, appeared to be either self-limited viral pneumonias or secondary bacterial or fungal pneumonias. Community respiratory viruses are frequent causes of acute respiratory illnesses in adult BMT recipients hospitalized during the winter and are associated with substantial morbidity and mortality. PMID- 8722931 TI - Bone and joint infections in patients with infective endocarditis: review of a 4 year experience. AB - Musculoskeletal complaints have been reported to occur in as many as 44% of patients with infective endocarditis (IE). A 4-year retrospective review of 104 cases of IE in which new diagnostic criteria were used showed that 24 episodes (23%) began with musculoskeletal complaints. Sixteen of the 24 episodes (66%) involved osteoarticular infection (OAI) documented during the period of hospitalization. Thus, in 15% of the 104 cases of IE, OAI was documented. The 16 episodes occurred in 15 patients, all of whom were intravenous drug abusers (IVDAs). In contrast, OAI was documented in none of the 38 cases in the IE population that did not involve IVDAs. Only three episodes were associated with single rather than multiple bone or joint involvement. There were no deaths, and the patients' conditions seemed to respond well to appropriate therapy. OAIs appear to be uncommon in patients with IE who are not IVDAs. Musculoskeletal complaints in the IVDA population with IE, however, should prompt a careful search for OAI. PMID- 8722932 TI - Treatment of erythema migrans in pregnancy. AB - The course of the illness and the outcome of pregnancy were investigated in a prospective study of 58 consecutively enrolled pregnant women with typical erythema migrans at the University Medical Centre of Ljubljana, Slovenia. Thirteen patients (22.4%) acquired borrelial infection in the first trimester of pregnancy, 27 (46.6%) in the second trimester, and 18 (31%) in the third trimester. All patients were treated for 14 days: 2 (3.4%) with phenoxymethylpenicillin (1 million IU thrice per day), 3 (5.2%) with benzylpenicillin (10 million units twice per day), and 53 (91.4%) with ceftriaxone (2 g daily). For 51 of the 58 women (87.9%), the pregnancies were normal and the infants were born at term, were clinically healthy, and had normal psychomotor development. Of the remaining 7 pregnancies, 1 ended with a missed abortion and 5 with preterm birth; 1 of the preterm babies had heart abnormalities. One child born at term was found to have urologic abnormalities at 7 months of age. A causal relationship between borrelial infection and fetal adverse effects has not been proven, and for at least some such cases a reasonable explanation (not associated with Lyme borreliosis) was found. PMID- 8722933 TI - Bacillary angiomatosis and bacillary peliosis in patients infected with human immunodeficiency virus: clinical characteristics in a case-control study. AB - Clinical characteristics associated with bacillary angiomatosis and bacillary peliosis (BAP) in patients with human immunodeficiency virus (HIV) infection were evaluated in a case-control study; 42 case-patients and 84 controls were matched by clinical care institution. Case-patients presented with fever (temperature, > 37.8 degrees C; 93%), a median CD4 lymphocyte count of 21/mm3, cutaneous or subcutaneous vascular lesions (55%), lymphadenopathy (21%), and/or abdominal symptoms (24%). Many case-patients experienced long delays between medical evaluation and diagnosis of BAP (median, 4 weeks; range, 1 day to 24 months). Case-patients were more likely than controls to have fever, lymphadenopathy, hepatomegaly, splenomegaly, a low CD4 lymphocyte count, anemia, or an elevated serum level of alkaline phosphatase (AP) (P < .001). In multivariate analysis, a CD4 lymphocyte count of < 200/mm3 (matched odds ratio [OR], 9.9; P < .09), anemia reflected by a hematocrit value of < 0.36 (OR, 19.7; P < .04), and an elevated AP level of > or = 2.6 mukat/L (OR, 23.9; P < .05) remained associated with disease after therapy with zidovudine was controlled for. BAP should be considered an AIDS-defining opportunistic infection and should be included in the differential diagnosis for febrile, HIV-infected patients with cutaneous or osteolytic lesions, lymphadenopathy, abdominal symptoms, anemia, or an elevated serum level of AP. PMID- 8722934 TI - Bacillary angiomatosis and parenchymal peliosis: frequent absence of mucocutaneous lesions. PMID- 8722935 TI - Vertical and horizontal transmission of unique Candida species to premature newborns. AB - The number of nosocomial bloodstream infections due to Candida species in critically ill newborns is increasing. This pathogen may be vertically transmitted from the mother or nosocomially acquired in the nursery. The goal of this study was to identify the route of transmission of unique Candida species and strains from mothers to their preterm offspring. Specimens from mothers for fungal cultures were obtained before delivery, and specimens from infants for sequential fungal cultures were obtained at defined intervals. Candida species were identified by standard methods and were typed by electrophoretic karyotyping (EK) and restriction endonuclease analysis of genomic DNA (REAG) with pulsed field gel electrophoresis. Antifungal susceptibility testing was performed on all isolates. Fungal cultures were positive for Candida species in 12 (63%) of 19 mothers' specimens and in seven (33%) of 21 infants' specimens. EK and REAG revealed that both the mother and the infant in three (14%) of 21 mother-infant pairs were colonized with the identical strain of Candida albicans. C. albicans was most commonly transmitted vertically. Candida parapsilosis colonized other infants and could not be accounted for by a maternal reservoir. PMID- 8722936 TI - Babesiosis in patients with AIDS: a chronic infection presenting as fever of unknown origin. AB - Babesiosis is a malaria-like, tick-transmitted zoonosis caused by protozoa of the family Piroplasmorida, which includes Babesia and Theileria species. In the United States, the infection is endemic in the Northeast and upper Midwest, although cases have recently been described in Northern California and Washington State. We report a case of babesiosis in a patient infected with HIV who presented with a prolonged fever of unknown origin; the patient had not undergone splenectomy. Parasitemia persisted despite initial clinical improvement after treatment with quinine and clindamycin. Babesiosis was controlled with a maintenance regimen consisting of clindamycin, doxycycline, and high-dose azithromycin, but the infection was not eradicated. Babesiosis should be considered in the differential diagnosis of HIV-infected patients with fevers and/or anemia in areas where the infection is endemic. HIV-infected patients who are severely immunosuppressed, even those without a history of splenectomy, may present with severe manifestations of babesiosis and develop a chronic infection, which may require therapy to prevent relapse of disease. PMID- 8722937 TI - Comparison of vancomycin, teicoplanin, metronidazole, and fusidic acid for the treatment of Clostridium difficile-associated diarrhea. AB - We conducted a prospective, randomized study to compare the efficacy of oral fusidic acid, oral metronidazole, oral vancomycin, and oral teicoplanin for the treatment of Clostridium difficile-associated diarrhea. Treatment resulted in clinical cure for 94% of the patients who were treated with vancomycin, 96% of those treated with teicoplanin, 93% of those treated with fusidic acid, and 94% of those treated with metronidazole. Clinical symptoms recurred in 16% of patients treated with vancomycin, 7% of those treated with teicoplanin, 28% of those treated with fusidic acid, and 16% of those treated with metronidazole. There was asymptomatic carriage of C. difficile toxin in 13% of patients treated with vancomycin, 4% of those treated with teicoplanin, 24% of those treated with fusidic acid, and 16% of those treated with metronidazole. No adverse effects related to therapy with vancomycin or teicoplanin were observed. Considering the costs of treatment, our findings suggest that metronidazole is the drug of choice for C. difficile-associated diarrhea and that glycopeptides should be reserved for patients who cannot tolerate metronidazole or who do not respond to treatment with this drug. PMID- 8722938 TI - Chlamydia pneumoniae infection in patients with cystic fibrosis. AB - The prevalence of atypical community-acquired infections as acute pulmonary exacerbations in patients with cystic fibrosis was prospectively studied. Thirty two patients admitted to the hospital because of acute pulmonary exacerbations and 24 clinically stable patients seen for their routine visits were examined. The prevalence of infection with Chlamydia pneumoniae was assessed by culture and serology, and the presence of IgE to C. pneumoniae was studied by immunoblotting. A subgroup of patients was also examined for the presence of Mycoplasma pneumoniae infection. C. pneumoniae was isolated from four patients presenting with acute pulmonary exacerbations (12.5%) and from none of the stable patients; all patients for whom cultures were positive also had IgE to C. pneumoniae. Polymerase chain reaction analysis for M. pneumoniae was not positive for any patient, and only one patient with an acute exacerbation had an antibody titer compatible with a recent infection. We conclude that infection with C. pneumoniae is associated with acute pulmonary exacerbations in some patients with cystic fibrosis and that it may trigger the production of IgE specific to C. pneumoniae, thus leading to bronchial reactivity in these patients. PMID- 8722939 TI - Evidence against person-to-person transmission of hantavirus to health care workers. AB - Unusual, primarily pulmonary, manifestations of hantaviral illness occurring in the southwestern United States raised the possibility of person-to-person transmission of a recently recognized hantavirus, Sin Nombre virus. To determine whether such transmission had occurred among health care workers (HCWs) exposed to patients with confirmed hantavirus pulmonary syndrome, we evaluated HCWs who had cared for patients with hantavirus pulmonary syndrome or who had processed specimens from these patients. Information about exposure to these patients and about recent illnesses was obtained via a standardized questionnaire. Serum specimens were tested for IgM and IgG antibodies to hantaviruses with use of ELISAs. Of the 396 HCWs, 266 (67%) reported that they had been exposed to patients with hantavirus pulmonary syndrome or to their body fluids or that they had processed laboratory specimens from these patients. Although 108 (27%) of the HCWs reported fever, myalgias, or respiratory illnesses during the 3 months before the serum specimens were obtained, hantavirus antibodies were not detected in any HCW. These data suggest that person-to-person transmission of Sin Nombre virus is unlikely to occur in health care settings. PMID- 8722941 TI - Disseminated intravascular coagulation associated with rotavirus gastroenteritis: report of two cases. AB - During one winter season, two children with rotavirus gastroenteritis who developed fulminant disseminated intravascular coagulation were seen at our hospital. Disseminated intravascular coagulation probably resulted from hypovolemic shock and acidosis, although extraintestinal spread of the virus cannot be excluded. PMID- 8722940 TI - Randomized controlled trial of a drug regimen that includes ciprofloxacin for the treatment of pulmonary tuberculosis. AB - The fluoroquinolones are promising new antituberculous agents. A randomized controlled trial of 200 adult patients with sputum smear-positive pulmonary tuberculosis was conducted in Tanzania. Patients received either a trial regimen (HRC) consisting of isoniazid (300 mg), rifampin (600 mg), and ciprofloxacin (750 mg) or a control regimen (HRZE) consisting of isoniazid (300 mg), rifampin (600 mg), pyrazinamide (25 mg/kg), and ethambutol (15 mg/kg). The 168 evaluable patients all had negative smears and cultures by month 6, but the time to conversion to negativity was longer for the HRC group than for the HRZE group because of the poor response of patients infected with human immunodeficiency virus (HIV) to the HRC regimen. Relapse was more frequent in the HRC group. The sterilizing activity of ciprofloxacin does not appear to be equal to that of the combination of pyrazinamide and ethambutol, but the difference in outcome was significant only among HIV-infected patients. PMID- 8722942 TI - High-dose fluconazole therapy for cryptococcal meningitis in patients with AIDS. AB - Fluconazole (800-1,000 mg i.v.) was administered to 14 consecutive patients with AIDS and cryptococcal meningitis. At 10 weeks the rate of clinical success was 54.5% (six of 11 patients responded to fluconazole); the Kaplan-Meier estimate of the response rate was 67.1%, and the overall mortality rate was 18.2% (two of 11 patients died). At the end of treatment, eight (72.7%) of 11 patients responded to fluconazole. The median time to the first negative cerebrospinal fluid (CSF) culture was 33.5 days (95% confidence interval, 18.3-67.3); the median time for patients with initial CSF cryptococcal antigen titers of > or = 1:1,024 was 66 days compared with 18 days for patients with initial CSF cryptococcal antigen titers of < 1:1,024 (P = .06). The median time to the first negative CSF culture for patients with an isolate for which the minimum inhibitory concentration (MIC) was 4 micrograms/mL was 56 days compared with 16 days for patients with an isolate for which the MIC was < 4 micrograms/mL (P = .11). The mean serum and CSF levels of fluconazole at steady state were 42.47 +/- 26.31 micrograms/mL and 36.63 +/- 21.08 micrograms/mL, respectively (ratio of CSF:serum, 0.86). No treatment was interrupted and no dose was tapered because of side effects. High dose fluconazole might be an effective and well-tolerated therapeutic option for patients with AIDS and acute cryptococcal meningitis. PMID- 8722944 TI - Pyogenic infections due to Ochrobactrum anthropi. AB - Ochrobactrum anthropi is a nonfermentative gram-negative bacillus that has been isolated with increasing frequency from human clinical specimens. Previously, its pathogenic niche was believed to involve the causation of catheter-associated bacteremic illnesses. We describe three cases of pyogenic infection due to O. anthropi, thereby expanding the known pathogenic potential of this organism. PMID- 8722943 TI - Occupational exposure leading to hantavirus pulmonary syndrome in a utility company employee. AB - Hantavirus pulmonary syndrome is a newly recognized rodent-borne zoonosis. We report a case of hantavirus pulmonary syndrome in an employee of a California utility company who was probably occupationally exposed to Sin Nombre virus. Environmental assessment and genetic comparison of the patient's hantavirus isolates to hantavirus isolates from rodents trapped at possible sites of exposure suggested that the patient contracted his infection at the work site. The study revealed a close correspondence between the patient's viral genotype and that from a rodent trapped at the work site. This report alerts the public health and medical community to the fact that employees of utility companies and similar industries may be an important risk group in areas where hantavirus is endemic and emphasizes the need to incorporate strategies for preventing exposure to hantavirus and other emerging infections into occupational safety protocols. PMID- 8722946 TI - A necrotic soft-tissue lesion due to Corynebacterium urealyticum in a neutropenic child. AB - Corynebacterium urealyticum has been associated mainly with infections of the urinary tract. Other infections due to this organism are highly unusual. We report what we believe is the first case of necrotic infection of soft tissue due to C. urealyticum in a neutropenic child who was previously treated with chemotherapy. The infection was cured when the patient was treated with vancomycin and surgical debridement. The increase in the number of neutrophils may also have contributed to the patient's recovery. PMID- 8722945 TI - Severe, protracted intestinal cryptosporidiosis associated with interferon gamma deficiency: pediatric case report. AB - We describe a human immunodeficiency virus-negative infant with chronic intractable diarrhea due to Cryptosporidium parvum; he had a history of significant weight loss since the second week of life. A preliminary study of his immune function was negative for primary immunodeficiency. To further study his immune function, proliferation of peripheral blood mononuclear cells (PBMCs) was elicited by crude antigenic extract from C. parvum oocysts (CCE). Supernatants of CCE-stimulated PBMCs contained interleukin 10 but not interferon gamma (IFN gamma), while PBMCs from a control (a 2-year-old immunocompetent infant who had recovered from cryptosporidiosis) showed a strong response to IFN-gamma when stimulated with CCE. These results stress the role of IFN-gamma in recovery from cryptosporidiosis. PMID- 8722947 TI - Infectious endocarditis due to non-typhi Salmonella in patients infected with human immunodeficiency virus: report of two cases and review. AB - Endocarditis is not usually considered a complication of AIDS. Because salmonellal bacteremia is common in HIV-infected patients and because salmonellae have a propensity to adhere to endothelial cells, these patients are at risk of endocarditis and endarteritis. We report two cases of endocarditis due to Salmonella enteritidis and review three previously reported cases. All five patients had underlying heart valve disease and developed fever, breakthrough or relapsing bacteremia, heart murmurs, and cardiac failure; four of five patients were older than 45 years. One patient died, but the other four were successfully treated with beta-lactam agents alone or in combination with aminoglycosides or with ofloxacin (valve replacement was not required). As AIDS patients get older, the number of cases of endocarditis or endarteritis due to Salmonella species may increase, particularly in geographic areas where Salmonella species are prevalent. PMID- 8722948 TI - Primary Vibrio damsela septicemia. PMID- 8722949 TI - Acute abdomen and intestinal necrosis produced by varicella-zoster virus in an immunocompromised host. PMID- 8722950 TI - Pyomyositis in intravenous drug abusers: report of a unique case and review of the literature. PMID- 8722951 TI - Fatal pneumococcal septic shock in a patient with ulcerative colitis. PMID- 8722952 TI - Pyomyositis as the sole manifestation of disseminated gonococcal infection: case report and review. PMID- 8722953 TI - Vertebral osteomyelitis due to Shigella boydii. PMID- 8722954 TI - Outer retinal necrosis due to a strain of varicella-zoster virus resistant to acyclovir, ganciclovir, and sorivudine. PMID- 8722955 TI - Age distribution among patients at high risk for human T-cell lymphotrophic virus type I infection. PMID- 8722956 TI - Clostridium perfringens as a cause of infectious endocarditis in a patient with a vascular prosthesis. PMID- 8722957 TI - Development of Aeromonas hydrophila bacteremia in a patient recovering from cholera. PMID- 8722958 TI - Ciprofloxacin- and azithromycin-resistant Campylobacter causing traveler's diarrhea in U.S. troops deployed to Thailand in 1994. PMID- 8722959 TI - Bacille Calmette-Guerin (BCG) meningitis in an AIDS patient 12 years after vaccination with BCG. PMID- 8722961 TI - Baker's cyst as a clinical presentation of brucellosis. PMID- 8722960 TI - Pulsatile chest wall swelling due to tubercular mediastinal lymphadenopathy in a patient with human immunodeficiency virus infection and syphilis. PMID- 8722962 TI - Tuberculosis-related retinal vasculitis in an immunocompetent patient. PMID- 8722963 TI - Bacteremia and hemoptysis due to an infected false left ventricular aneurysm associated with a bronchoventricular fistula. PMID- 8722964 TI - Staphylococcal toxic shock syndrome associated with human immunodeficiency virus infection: report of a case with bacteremia. PMID- 8722965 TI - Levels of nitric oxide correlate with high levels of tumor necrosis factor alpha in cerebrospinal fluid samples from children with bacterial meningitis. PMID- 8722966 TI - Reversible peripheral neuropathy associated with sodium stibogluconate therapy for American cutaneous leishmaniasis. PMID- 8722967 TI - Anaerobic septic bursitis: case report and review. PMID- 8722968 TI - Vincristine as therapy for idiopathic thrombocytopenic purpura in patients infected with human immunodeficiency virus. PMID- 8722969 TI - Recurrent erysipelas or erysipelas-like rash? PMID- 8722970 TI - Progressive multifocal leukoencephalopathy as an AIDS-defining condition in a patient with human immunodeficiency virus infection. PMID- 8722971 TI - Levels of interleukin-10 and tumor necrosis factor alpha in patients with bacterial meningitis. PMID- 8722972 TI - Adverse events associated with high-dose rifabutin in macrolide-containing regimens for the treatment of Mycobacterium avium complex lung disease. PMID- 8722973 TI - Use of famciclovir and valaciclovir in the treatment of viral keratitis. PMID- 8722974 TI - Invasive staphylococcal infection complicating coronary angiography without angioplasty. PMID- 8722975 TI - Craniofacial injuries resulting from taxicab accidents in New York City. AB - Taxicab accidents are a common occurrence in New York City. This review was undertaken to characterize the nature of craniofacial injuries that result from taxicab accidents. Data were collected on 16 patients who required admission to trauma or plastic and reconstructive surgery services, after sustaining craniofacial injury as a result of a taxicab accident. Front-end deceleration collisions were the most common mechanism of injury. Fifty-six percent of the patients were thrown against the bulletproof, Plexiglas driver safety divider and sustained an injury most commonly to the anterior midface. Both bony and soft tissue injuries were common in the entire group. Complex facial fractures were sustained by 56% of patients, with nasal-septal fractures most common, followed by naso-ethmoid-orbital, anterior frontal, anterior maxillary, and Le Fort I and II fractures. Only 1 patient in the group was wearing a seat belt and that patient was a driver. Given the high incidence of craniofacial injury, appropriate safety standards for taxicabs must be initiated, including the reevaluation of the utility of the safety divider and mandatory seat belt use for rear-seat passengers. PMID- 8722976 TI - Hemostasis at skin graft donor sites: evaluation of topical agents. AB - Blood loss from split-thickness skin graft donor sites may be significant. Various topical agents have been used to decrease this blood loss, including thrombin and epinephrine solutions of varying concentrations. We describe a K-Y jelly/epinephrine mixture that serves both as a lubricant for the dermatome and as a hemostatic agent. This mixture, in comparison with other topical agents, produces rapid hemostasis and offers the advantages of easy use, ready availability, and low cost. The blood loss savings based on this hemostatic technique is quantifiable and significant. PMID- 8722977 TI - Warmed local anesthetic reduces pain of infiltration. AB - The effect of warming local anesthetic on the amount of pain experienced during local infiltration was tested by comparing equal volumes of 40 degrees C- and 21 degrees C-infiltrates in each of 26 subjects. Six subjects were patients undergoing excision of two benign asymptomatic nevi in separate locations, and 20 subjects were healthy adult volunteers who were injected in bilateral antebrachial sites. The warmed and room temperature solutions were randomized to each side, so that each subject received both temperature injections in random order. All subjects and the injector were blinded. The rate of injection was time controlled (0.05 ml/sec). Following both injections, subjects were asked to rate the pain experienced at each site. In addition, the subject was asked if there was no difference, a slight difference, or a substantial difference between the two sites. A two-tailed paired t-test was used to analyze the mean difference in pain scores for all subjects. Paired analysis of the pain scores for each subject eliminated intersubject variance of pain tolerance. The mean difference in pain score between the room temperature and warmed solutions was +1.5 (p < 0.0001). Of the 21 subjects (81%) who found the warmed solution less painful, 11 (52%) found the difference to be significant, while 10 (48%) found the difference to be slight. Two subjects (8%) found no difference between the two, while 3 subjects (11%) found the colder solution slightly less painful. We conclude that warming local anesthetic to 40 degrees C prior to subcutaneous injection is a simple, inexpensive means of reducing the pain of local infiltration. PMID- 8722978 TI - Arterial thrombosis enhanced by an acute phase response. AB - Induction of an acute phase response causes changes in the levels of specific plasma proteins. Fibrinogen is elevated to approximately twice its normal concentration by trauma, inflammation, or infection. In this study, an acute phase response was induced by subcutaneous turpentine injection in rats. Twenty four hours later, an arterial model of thrombosis was created bilaterally in the femoral arteries. Patency at 1 and 7 days postoperatively was 24% (9/38) in comparison to 56% (20/36) in control (saline-injected) rats undergoing the same thrombosis model. Fibrinogen levels were elevated in the turpentine group to 5.3 +/- 0.8 mg/ml at the time of the surgery and 7.2 +/- 0.3 mg/ml 24 hours after surgery. In contrast, the control group had plasma fibrinogen levels of 2.5 +/- 0.2 mg/ml at the time of surgery and 5.4 +/- 0.5 mg/ml 24 hours postoperatively. These findings suggest that when an acute phase reaction has been induced several hours prior to a microvascular procedure, there may be an increased risk for development of arterial thrombosis. PMID- 8722979 TI - High-energy gunshot wounds to the face. AB - Thirty-three patients with high-energy gunshot wounds to the face were treated at the University of Kentucky Chandler Medical Center between 1976 and 1993. Wounds were classified according to the mass and velocity of the projectile, and the range from weapon to target. More than half the injuries involved multiple facial regions. Twenty patients underwent immediate definitive reconstructive procedures. Intervals between injury and initial nondefinitive reconstruction for the other patients ranged from 1 day to 1 month. Toward the end of the study period, reconstruction was undertaken earlier and more aggressively, and included more attention to primary bone grafting and free tissue transfer. These patients developed fewer problems with infection, long-term scarring, and contracture, and they required fewer operative procedures. There was no operative mortality and none of the patients with self-inflicted injuries reattempted suicide. We conclude that early aggressive treatment of these wounds can produce better structural, functional, and rehabilitative results. PMID- 8722980 TI - Upper airway obstruction after pharyngeal flap surgery. AB - Upper airway obstruction after superiorly based pharyngeal flap procedures for the treatment of velopharyngeal insufficiency is described in this series of 32 flaps performed in 29 patients at our institution between 1979 and 1993. A high incidence of upper airway obstruction symptoms (38%) occurred in the early postoperative period but resolved in all but 2 patients within 5 months. None of the patients required flap revision or other procedures for relief of upper airway obstruction. Velopharyngeal insufficiency was improved or completely eliminated in 87% of patients. Significant residual velopharyngeal insufficiency in 13% of patients was treated successfully in all flap revision cases. Race, gender, age at closure, and associated craniofacial anomalies did not correlate with upper airway obstruction or with the efficacy of treatment for velopharyngeal insufficiency. However, patients with transient postoperative upper airway obstruction were only half as likely to suffer residual postoperative velopharyngeal insufficiency. The inverse relationship between velopharyngeal insufficiency and upper airway obstruction (p = .008) suggests that the surgeon may sometimes need to accept some transient upper airway obstruction to achieve correction of velopharyngeal insufficiency. PMID- 8722981 TI - Anatomical parameters for nipple position and areolar diameter in males. AB - In this paper we present anatomic parameters for nipple position and areolar diameter in males. Larger forms of gynecomastia with significant ptosis pose a challenge to the plastic surgeon with respect to relocation of the nipples on the chest wall. Selection of the appropriate areolar size is also of concern in gynecomastia correction. There is a paucity of information in the current literature pertaining to this problem. In order to establish guidelines for the placement of the nipple in gynecomastia correction and for the selection of the appropriate areolar size, we set out to determine these anatomic parameters. We believe use of these parameters will enhance the aesthetic results of gynecomastia correction. One hundred males between the ages of 17 to 30 years were chosen for this study. The males selected were of ideal body weight and without evidence of gynecomastia. The distances from the sternal notch to the nipple, the midclavicular line to the nipple, and the nipple-to-nipple distance were recorded. The areolar diameter was also measured in each subject. The average distances were determined for each category. The validity of these values was confirmed with statistical analysis. Equations were then derived, using this analysis, to determine nipple position in males. We have determined the nipple position in males to be approximately 20 cm from the sternal notch and 18 cm from the midclavicular line. The ideal nipple-to-nipple distance is 21 cm. The average areolar diameter is 2.8 cm. PMID- 8722982 TI - Microsurgical reconstruction of the head and neck: interdisciplinary collaboration between head and neck surgeons and plastic surgeons in 305 cases. AB - Three hundred five microsurgical free flaps have been performed for defects of the head and neck by a team of two head and neck surgeons and two plastic surgeons over a 9-year period, with a success rate of 91.2%. The most common flaps used were the jejunum (89), radial forearm (57), rectus abdominis (48), latissimus dorsi (40), scapular (32), fibula (15), and iliac crest (11). Thirty three flaps required reexploration for anastomotic thrombosis or hematoma (10.8%), of which 18 flaps were salvaged (54.5%). Thirteen flap failures occurred in 113 patients who had received preoperative irradiation (11.5%), but this was not statistically significant. Seven flaps failed in 20 patients who required an interposition vein graft (35%) and this was statistically significant. Ninety patients (31.5%) developed a major complication other than anastomotic thrombosis or death. Despite postoperative intensive care nursing and monitoring, 18 patients died postoperatively in the hospital (6.3%). The average hospital stay was 21.1 days with a range from 5 to 95 days. During this 9-year time period, various free flaps have evolved as the preferred choice for free flap reconstruction of a specific defect of the head and neck. The latissimus dorsal muscle flap surfaced with a nonmeshed split-thickness skin graft is the optimal free flap for reconstruction of the scalp and skull, whereas a multiple-paddle latissimus dorsi musculocutaneous flap is the best flap for reconstruction of complex defects of the middle third of the face and maxilla. The radial forearm flap and free jejunal transfer have become the preferred choices for intraoral reconstruction and pharyngo-esophageal reconstruction, respectively. There still remains no universally accepted flap for mandibular reconstruction, but the fibular osteocutaneous flap and a reconstruction plate protected by a radial forearm flap have largely superseded the iliac crest and scapular osteocutaneous flaps. Radical resection of tumors of the head and neck with immediate reconstruction by microsurgical free tissue transfer followed by adjuvant radiation therapy provides the best possible chance for cure and functional and social rehabilitation of the patient. PMID- 8722983 TI - Absorbable and nonabsorbable buried sutures for primary cleft lip repair. AB - Absorbable and nonabsorbable buried sutures were studied in primary cleft lip repair. Group 1 (N = 56) consisted of patients repaired with buried nonabsorbable material (monofilament nylon). Group 2 (N = 47) consisted of patients repaired with absorbable materials (polyglyconate, polydioxanone). All patients were monitored for 12 months. There were stitch abscesses in Group 1 (14%). There were no abscesses in Group 2. This difference was significant (p = 0.007). Abscesses were located in the suture line with no identifiable distribution. There was no significant difference in the cosmetic appearance of the scars in Groups 1 and 2. These results support the view that absorbable sutures are preferable to nonabsorbable sutures for primary cleft lip repair. PMID- 8722984 TI - A reposition of Tennison's triangular flap in long lip deformity. AB - The authors have used a new technique in 5 cases with severe long lip deformity and drooping of the Cupid's bow, who all have undergone Tennison's triangular flap method for unilateral cleft lip at primary repair. We corrected this deformity by repositioning Tennison's triangular flap back into its original site and then obtained (1) effective shortening of a vertical lip length, (2) restoration of a symmetric Cupid's bow, (3) prevention of tight upper lip, (4) refinement of the transverse scar, and (5) low recurrence. PMID- 8722985 TI - Multiple segmental gradual distraction of facial skeleton: an experimental study. AB - The method of distraction osteogenesis was used to move several facial segments in different directions with formation of new bone. On 3 young adult sheep a Le Fort II osteotomy, combined with circumferential orbital osteotomies, was performed. The periosteum over the osteotomies was preserved. Titanium bone screws were placed as bone markers. The segments were fixed with pins and plates to a combined distraction apparatus in order to advance the segments in different directions. By a gradual distraction, the midface was advanced forward by 30 mm. The left orbit was moved laterally 20 mm and the right orbit was moved forward 20 mm. In both orbits there was a concomitant globe movement. New bone formation in the distracted area was obvious radiographically, clinically, and histologically. In conclusion, multiple segmental distraction is possible in the adult sheep face and may offer three-dimensional (3D), controlled correction of complex face deformities. PMID- 8722986 TI - Healing of microvenous polytetrafluoroethylene (PTFE) prostheses implanted into the rat femoral vein by means of 3M precise. AB - Thirty PTFE prostheses (internal diameter, 1.0 mm; length, 5.0 mm; fibril length, 30 microns) were implanted into rats' femoral veins by means of a coupling device (3M precise microvascular anastomosis system, 3M, St Paul, MN) and evaluated with electron microscopy at regular intervals from 1 day to 3 weeks after implantation to study, in detail, the healing process. Eighty-three percent of mechanically anastomosed grafts were found to be patent. At 1 and 3 days after implantation, the whole length of PTFE was covered with a clot layer containing platelets and a fibrin network. After 1 week, endothelial-like cells originating from the anastomotic sides grew in across the anastomoses. At 3 weeks, the prostheses were completely covered by an endothelial-like cell layer. These results demonstrate that the degree of neo-endothelialization in the microvenous PTFE prosthesis anastomosed with 3M rings was not delayed, as was seen in the healing of microarterial PTFE tubing that was mechanically anastomosed. PMID- 8722987 TI - Using hyaluronic acid to create a fetal-like environment in vitro. AB - The fetal wound healing matrix is exceptionally rich in hyaluronic acid (HA). Fetal wounds heal without scarring or contraction. Noting these observations, we cultured adult dermal explants in the presence of various concentrations of medical-grade HA in vitro. In the presence of HA, fibroblasts migrated from the dermal explant and multiplied more rapidly than control explants. Subsequently, sterile toothpicks were used to disrupt (wound) fibroblast monolayers mechanically and the rate of closure was monitored. Cells cultivated in the presence of 5 mg/ml of exogenous HA changed in morphology and closed the wound more quickly than control cultures. Cells surrounding the wound extended numerous podalic processes and showed increased interdigitation. The effect of HA on cell proliferation is usually discussed in terms of the mechanical effects HA exerts on cells and the extracellular matrix. The physiological effect of HA may lie in its ability to act as an accessory receptor in cooperative ligand-binding pathways. For example, HA may bind growth and/or other factors, and thereby increase the effective concentration of these factors at the cell surface. PMID- 8722988 TI - Soft-tissue blood flow following reamed versus unreamed locked intramedullary nailing: a fractured sheep tibia model. AB - A study was performed to compare the effects of reamed versus unreamed locked intramedullary nailing on soft-tissue blood flow in a fractured sheep tibia model. After creation of a standardized short spiral fracture, each tibia was stabilized by insertion of a locked intramedullary nail. Ten animals were randomized into two groups: one that had reaming prior to nail insertion and one that did not. Blood flow was measured in real time, using laser Doppler flowmetry. Skin and muscle perfusion measurements were made at three locations (proximal diaphysis, fracture site, and distal diaphysis) and at five time intervals (postfracture; postnail insertion; and at 2-week, 6-week, and 12-week follow-ups). All animals were sacrificed at 12 weeks postoperatively. Muscle blood flow remained elevated longer in the reamed group than in the unreamed group, following completion of the procedure. Muscle perfusion was significantly greater in the reamed group than in the unreamed group (p < 0.033) and was significantly greater than skin perfusion at 2 weeks (p < 0.024). There was no difference in skin perfusion between the reamed and unreamed groups at any time interval. The study demonstrates that muscle perfusion is altered following reamed and unreamed intramedullary nailing. The findings demonstrate the change in blood flow in the surrounding soft-tissue envelope during the process of revascularization, following devascularization of a segment of tibial cortex by intramedullary nailing. PMID- 8722989 TI - Chronic, burning facial pain following cosmetic facial surgery. AB - Chronic, burning facial pain as a result of cosmetic facial surgery has rarely been reported. During the year of 1994, two female patients presented themselves at our Pain Relief Clinic with chronic facial pain that developed following aesthetic facial surgery. One patient underwent bilateral transpalpebral surgery for removal of intraorbital fat for the correction of the exophthalmus, and the other had classical face and anterior hairline forehead lifts. Pain in both patients was similar in that it was bilateral, symmetric, burning in quality, and aggravated by external stimuli, mainly light touch. It was resistant to multiple analgesic medications, and was associated with significant depression and disability. Diagnostic local (lidocaine) and systemic (lidocaine and phentolamine) nerve blocks failed to provide relief. Psychological evaluation revealed that the two patients had clear psychosocial factors that seemed to have further compounded their pain complaints. Tricyclic antidepressants (and biofeedback training in one patient) were modestly effective and produced only partial pain relief. PMID- 8722990 TI - Treatment of chronic, nonhealing abdominal wound in a liquid environment. AB - A 66-year-old woman with an abdominal wound caused by infected synthetic mesh had failed to heal in spite of many surgical attempts to close the wound. A sealed transparent vinyl chamber was glued to the periphery of the wound and antibiotics in high concentrations (up to 2,500 times the minimum inhibitory concentration) were delivered through the chamber. The wound fluid in the chamber was used for analysis of microbial activity, concentration of residual antibiotics, and growth factor activity. After 10 weeks of treatment, the wound was closed and has not recurred in 24 months. PMID- 8722991 TI - Late rupture of a flexor tendon after electrical injury: tendon localization using magnetic resonance imaging. A case report. AB - Rupture of a flexor tendon in the nonrheumatoid population is rare. Localizing the level of the tendon rupture can be a difficult task. We describe an individual who presented with the sudden inability to flex the index finger of the hand, which had been subjected to a severe electrical injury. Physical exam demonstrated lack of index finger flexion at the distal joint. The patient denied any pain or history of recent trauma. Preoperative magnetic resonance imaging (MRI) successfully located the proximal and distal ends of the ruptured flexor digitorum profundus tendon in the forearm. MRI allows for accurate preoperative assessment of tendon position and degree of retraction, thereby facilitating surgical planning and approach. Accurate localization of tendon pathology preoperatively minimizes unnecessary dissection, shortens the operative procedure, and clarifies operative planning. PMID- 8722992 TI - Scarpa's adipofascial flap for repair of wide scalp defects. AB - Scarpa's fascia is a prominent superficial fascial system of the body. It consists of a single membrane between the superficial fatty layer and deep fatty layer, and lies widely in the lower abdominal wall. We describe a case with a wide scalp defect resulting from a resection of a dermatofibrosarcoma, and reconstruction of the defect with Scarpa's adipofascial flap (i.e., a combined paraumbilical perforator-based adipofascial flap-groin adipofascial flap). The primary advantage of Scarpa's adipofascial flap for scalp defects is that (1) the donor site is most acceptable for a free flap with a minimal donor scar and minimal dysfunction; (2) even in cases in which large flaps are used, donor defects can be closed directly without skin grafting; (3) in the obese patient, this flap is preferable because of cosmetic improvement of the abdominal wall; (4) the donor area has so many perforators that an extended adipofascial flap can be obtained with a combination of these perforators; and (5) the flap may be nourished with one of several arteries, such as the superficial or deep inferior epigastric artery, or the superficial or deep circumflex iliac artery. The disadvantages of this flap are that the territory with a single artery may be smaller than a skin flap with the same artery and oversurfacing of the graft results in a poor cosmetic appearance. Scarpa's adipofascial flap is indicated when the defects are in an exposed area, especially in children, young patients, and females, and when this procedure is combined with a skin-expanding method in the secondary repair. PMID- 8722993 TI - Above-elbow arm replantation during pregnancy. AB - We report a successful replantation of an above-elbow amputated left arm in a 26 year-old female with a good, functional, long-term result. The patient was at 18 weeks estimated gestational age at the time of surgery. Major upper extremity replantation during pregnancy has not been previously described in the literature. Current operative and perioperative aspects, including considerations that favored microsurgical replantation in this unusual case, are discussed. PMID- 8722994 TI - H-plasty: a new modification for correction of nasal stenosis. AB - Unilateral complete nasal stenosis is a rare condition and is usually acquired. The alar wing and nasal vestibule may be disturbed because of stenosis. We operated on 2 patients with complete nasal stenosis by using the H-flap technique described in this paper. Anterior nares are formed by this technique and, in one case, we used a cartilage graft for creating symmetry of the alar wings. Functional and aesthetic anterior nares are obtained by this technique. PMID- 8722995 TI - Familial polythelia without associated anomalies. AB - Of the many forms of supernumerary breast tissue, the most common form is the isolated presence of an accessory nipple, polythelia. While familial polythelia is recognized, it is extremely rare. In the past several years, polythelia has been noted to be associated with nephrourological anomalies. All reports of such a relationship are in random, nonfamilial cases of polythelia. We report three cases of polythelia in a family over two generations who had no urinary tract abnormalities. Discussion includes a comprehensive review of familial polythelia and its association with renal anomalies. From this review, the association of familial polythelia with nephrourological abnormalities will be delineated. Although in this report of a single family with polythelia we did not demonstrate any renal anomalies, we feel that a thorough physical exam, urine analysis, and renal ultrasound should be pursued in any patient with a significant familial history of polythelia. PMID- 8722996 TI - Supernumerary extensor tendon to the thumb: a case report. AB - Anomalies of the dorsal tendons of the hand are not uncommon. Numerous variations in the tendons coursing through the fourth dorsal extensor compartment have been described. We report a rare case of a supernumerary tendon passing through the fourth dorsal compartment and inserting into the distal phalanx of the thumb. We also review the anomalous muscles and tendons that are found in this compartment. PMID- 8722997 TI - Stable trait components of hopelessness: baseline and sensitivity to depression. AB - Hopelessness (H) plays an important theoretical and practical role in depression. The authors hypothesized that a patient's H is comprised of (a) a baseline level of H when not depressed and (b) an increment in H related to the severity of depression at the time and the person's rate of increase in H as a function of severity of depression (sensitivity). Baseline and sensitivity are explanatory stable traits; H and depression are observed, time-varying states. The corresponding statistical model described well the longitudinal data of 316 participants. Baseline and sensitivity were uncorrelated and correlated with different clinical and demographic variables. Baseline predicted a future suicide attempt; sensitivity and H when depressed did not. It may be useful to ask "How hopeless is this person when not depressed and how much more hopeless is he or she when depressed?", rather than simply "How hopeless is this depressed person?" PMID- 8722998 TI - Escalated substance use: a longitudinal grouping analysis from early to middle adolescence. AB - The authors surveyed a cohort of 1,184 adolescents in the 7th, 8th, and 9th grades. Measures of tobacco, alcohol, and marijuana use and of constructs from 3 theoretical models of substance use were obtained at each point. Clustering analysis for 3-wave substance use data indicated subgroups of nonusers, minimal experimenters, late starters, and escalators. Discriminant function analyses tested whether study variables differentiated the subgroups. One discriminant function accounted for the majority of between-group association; it had loadings for (high) life stress, nonadaptive coping, deviance-prone attitudes, and parental and peer substance use, and (low) parental support, academic competence, and behavioral control. Escalators were high on this function; late starters and experimenters had intermediate values; and nonusers were low on the function. Implications for theories of vulnerability to substance abuse are discussed. PMID- 8722999 TI - Structure of anxiety and the anxiety disorders: a hierarchical model. AB - The authors used a semistructured clinical interview and a self-report battery of questionnaires to measure key features of the anxiety disorders in a large sample of patients seeking treatment at an outpatient anxiety disorders clinic and in a no mental disorder group. Results were consistent with hierarchical models of anxiety and the anxiety disorders such as the model implicit in American Psychiatric Association (1987, 1994) and trait models positing a trait diathesis common to all the anxiety disorders. A higher order general factor differentiated each of the patient groups from the no mental disorder group. Several lower order factors provided the basis for differentiation among the patient groups. Conclusions regarding the degree to which models predicting a hierarchical structure of anxiety and the anxiety disorders are empirically supported must await replication of these results with additional samples. PMID- 8723000 TI - Genital and psychoaffective response to erotic stimulation in sexually functional and dysfunctional men. AB - To understand the sexual response patterns of men with premature ejaculation and erectile dysfunction, the authors compared genital and affective responses of sexually functional and dysfunctional men with 3 types of sexual stimulation: an erotic video, penile vibrotactile stimulation, and a combination of both. Genital response differed across both groups and stimulus conditions, with an interactive effect indicating that groups showed different response patterns depending on the stimulation. Affective responses also differed across groups and interacted with stimulus conditions. The combination genital and affective response was superior to either alone in distinguishing men with no sexual problems from those with erectile or ejaculatory problems (or both). These factors were particularly useful in discriminating men with premature ejaculation from those with combined premature ejaculation and erectile dysfunction. PMID- 8723001 TI - Memory performance among women with parental abuse histories: enhanced directed forgetting or directed remembering? AB - Performance on a directed forgetting task was assessed in 24 individuals with borderline personality disorder and early life parental abuse, 24 borderline individuals with no history of abuse, and 24 healthy nonclinical controls under conditions of explicit and implicit memory. In the explicit memory condition, individuals with abuse histories showed greater differential recall of "to-be remembered" versus "to-be-forgotten" material compared to the 2 comparison groups. Implicit memory performance was equivalent for all 3 groups. The enhanced selective memory in the abused group was the result of better recall for "remember" and not poorer recall for "forget" information, indicating that abused individuals have an enhanced ability to sustain attention to designated "remember" information. Because most people with childhood abuse recall their abuse, enhanced remembering of designated events (e.g., information not associated with abuse) may be a coping strategy. PMID- 8723002 TI - Working memory, attention, and communication disturbances in schizophrenia. AB - The authors hypothesized that schizophrenic communication disturbances reflect specific cognitive deficits in the areas of working memory and attention. They examined the cognitive correlates of communication disturbances, as measured by linguistic reference performance, in schizophrenic (n = 48), bipolar (n = 24), and nonpsychiatric control (n = 23) individuals. Reference performance ratings in the schizophrenic patients were associated with scores on tests of working memory and attention and were not related to performance on concept formation or verbal fluency tests. In contrast, in the bipolar and nonpsychiatric individuals, reference performance was associated with concept formation and verbal fluency test scores but was not related to performance on tests of working memory. Implications with respect to the processes underlying schizophrenic communication disturbances are discussed. PMID- 8723003 TI - Depressive symptomatology in new mothers: a stress and coping perspective. AB - The aim of the present research was to test the utility of a stress-coping model of postpartum depression. Data were collected during the last trimester of pregnancy (n = 197) and twice after the birth (4 weeks, n = 180, and approximately 5 months, n = 163). Coping resources and depressive symptomatology were assessed at Time 1, stress and coping were assessed at Time 2, and depressive symptomatology and partner ratings of coping effectiveness were assessed at Times 2 and 3. After control of the effects of initial depression, there was evidence of significant effects of levels of stress and coping responses on the Time 2 and Time 3 outcome measures. There were also some evidence linking coping resources (particularly self-esteem and family support) to postpartum depressive symptomatology. PMID- 8723004 TI - Appraisal of self, social environment, and state authority as a possible mediator of posttraumatic stress disorder in tortured political activists. AB - This study examined appraisal of self and others, as measured by semantic differential ratings of Police, State, Society, Family, Friend, Myself as a Man/Woman, and Myself as a Political Person, in 55 tortured political activists in Turkey, 55 nontortured political activists, and 55 nontortured, politically noninvolved controls. There were no remarkable differences between tortured and nontortured political activists; both groups differed from controls in having a more negative appraisal of the police and the state and stronger perceptions of danger, mistrust, and injustice in relation to state authority. Lack of beliefs concerning a "benevolent state" may have protected the survivors from the traumatic effects of state-perpetrated torture. Further research into the possible protective role of belief systems in posttraumatic stress disorder is needed. PMID- 8723005 TI - Prediction of posttraumatic stress symptoms in children after hurricane Andrew. AB - The authors used an integrative conceptual model to examine the emergence of posttraumatic stress disorder (PTSD) symptoms in 568 elementary school-age children 3 months after Hurricane Andrew. The model included 4 primary factors: Exposure to Traumatic Events, Child Characteristics, Access to Social Support, and Children's Coping. Overall, 62% of the variance in children's self-reported PTSD symptoms was accounted for by the 4 primary factors, and each factor improved overall prediction of symptoms when entered in the analyses in the order specified by the conceptual model. The findings suggest that the conceptual model may be helpful to organize research and intervention efforts in the wake of natural disasters. PMID- 8723006 TI - Do schizophrenic patients show a disjunctive relationship among expressive, experiential, and psychophysiological components of emotion? AB - Recent research has found a discrepancy between schizophrenic patients' outward expression of emotion and their reported emotional experience. In this study, which attempts to replicate and extend the findings of previous studies, participants with and without schizophrenia viewed emotional film clips while their facial expressions were videotaped and skin conductance was recorded. Participants also reported their subjective experience of emotion following each film. Those with schizophrenia were less facially expressive than controls during the emotional films and reported experiencing as much positive and negative emotion, replicating previous findings. Additionally, schizophrenic patients exhibited greater skin conductance reactivity to all films than controls. These findings suggest a disjunction among emotional response domains for schizophrenic patients; alternative explanations for the findings are considered as well as suggestions for future research. PMID- 8723007 TI - Modeling causal relations between academic and social competence and depression: a multitrait-multimethod longitudinal study of children. AB - The authors obtained self-reports, peer nominations, teacher ratings, and parent reports of depression and social and academic competence on 490 3rd graders and 455 6th graders near the beginning and end of the school year. Confirmatory factor analysis and structural equation modeling revealed that (a) measures showed significant convergent and discriminant validity; (b) within-wave correlations between constructs were large and significant, although the depression-social competence correlation was larger than the depression-academic competence correlation; (c) the cross-wave stability of all constructs was remarkably high; and (d) social competence at Wave 1 predicted depression at Wave 2 for 6th graders after controlling for depression at Wave 1. Depression did not predict change in either academic or social competence over time. Implications for competence-based and failure-based models of child depression are discussed. PMID- 8723008 TI - Emotion recognition and social competence in chronic schizophrenia. AB - This study evaluated (a) whether chronic, medicated schizophrenia patients show deficits in emotion recognition compared to nonpatients, and (b) whether deficits in emotion recognition are related to poorer social competence. Two emotion recognition tests developed by S. L. Kerr and J. M. Neale (1993) and Benton's Test of Facial Recognition (A. Benton, M. VanAllen, K. Hamsher, & H. Levin, 1978) were given to patients with chronic schizophrenia and nonpatient controls. Patients' social skills, social adjustment, and symptomatology were assessed. Like Kerr and Neale's unmedicated patients, these patients performed worse than controls on both emotion recognition tests and the control test. For patients, facial perception was related to the chronicity of illness and social competence. Chronicity of illness may contribute to face perception deficits in schizophrenia, which may affect social competence. PMID- 8723009 TI - Perfectionism and depression: longitudinal assessment of a specific vulnerability hypothesis. AB - The authors tested whether perfectionism dimensions interact with specific stress to predict depression over time. A sample of 103 current and former patients completed measures of perfectionism and depression at Time 1 and measures of stress and depression 4 months later. After controlling Time 1 depression, self oriented perfectionism interacted only with achievement stress to predict Time 2 depression. Socially prescribed perfectionism did not interact with achievement or social stress to predict Time 2 depression, but it did predict Time 2 depression as a main effect. The results provide support for the contention that perfectionism dimensions are involved in vulnerability to depression over time. PMID- 8723010 TI - Postpartum psychiatric disorders: their relationship to psychological adjustment and marital satisfaction in the spouses. AB - The authors examined mental health and marital quality in an index group of spouses of women with postpartum psychiatric disorders and a control group of men whose wives had recently given birth but had no such disorders. At 6 to 9 weeks postpartum, couples underwent a psychiatric interview and completed self-report measures of psychological symptoms, marital satisfaction, and changes in couple and family functioning since the birth. Index spouses reported more symptoms and had lower Global Assessment of Functioning (R. L. Spitzer, J. B. W. Williams, M. Gibbon, & M. B. First, 1990) scores than controls. Index men reported greater marital dissatisfaction and more change in household routines, recreation, and intimacy with their partners than controls. PMID- 8723011 TI - Anxiety, depression, and the anticipation of future positive and negative experiences. AB - An experiment is reported that attempts to distinguish between anxious and depressive future thinking in terms of anticipation of future positive and future negative experiences. Anxious, mixed (anxious-depressed), and control participants were given an adapted verbal fluency paradigm to examine the ease with which they could think of future positive and negative personal experiences. Anxious participants differed from controls only in anticipating more future negative experiences; mixed participants showed both greater anticipation of negative experiences and reduced anticipation of positive experiences. Self report measures of hopelessness and worry followed a similar pattern to future positive and future negative anticipation, respectively. The results are discussed in terms of the distinction between positive affect and negative affect (D. Watson, L. A. Clark, & G. Carey, 1988). PMID- 8723012 TI - Profibrinolytic and antifibrinolytic effects of platelets. AB - The pro- and antifibrinolytic effects of platelets have been reviewed periodically over the past 25 years. Review of the current publications indicates that the profibrinolytic and antifibrinolytic effects attributed to platelets are highly dependent upon the test system used. On balance, the antifibrinolytic effect appears to predominate; however, this conclusion awaits further experimentation, particularly through clinical studies clarifying the effects of platelets in therapeutic fibrinolysis. PMID- 8723013 TI - Platelet activation by fibrinolytic agents: a potential mechanism for resistance to thrombolysis and reocclusion after successful thrombolysis. PMID- 8723014 TI - Antiplatelet effects of fibrinolytic agents: a potential contributor to the hemostatic defect after thrombolysis. PMID- 8723015 TI - Aspirin augmentation of the efficacy of thrombolysis. PMID- 8723016 TI - Adjunctive therapy in coronary thrombolysis: prostaglandins, nitrates and calcium antagonists. PMID- 8723017 TI - Inhibitors of platelet glycoprotein IIb/IIIa as augmenters of thrombolysis. PMID- 8723018 TI - Improved beta-adrenergic receptor function after coronary artery bypass grafting in patients with congestive heart failure. AB - BACKGROUND: To evaluate whether coronary artery bypass grafting (CABG) in patients with congestive heart failure (CHF) modifies beta-adrenergic receptor dysfunction, we measured lymphocytic beta-adrenergic receptor density and function, and plasma catecholamine levels in patients with congestive ischaemic disease scheduled for CABG. METHODS: Assays were performed on 20 patients with CHF at admission and 6 months following CABG; 20 age-matched healthy subjects served as the control group. RESULTS: Preoperative lymphocyte beta-adrenergic receptor density was significantly reduced in patients with CHF compared with the control group. Similarly, basal adenylyl cyclase as well isoproterenol- and NaF stimulated adenylyl cyclase activity was significantly reduced in lymphocytic membranes of patients with CHF compared with controls. Conversely, no difference was found in forskolin-stimulated adenylyl cyclase activity between the two groups of patients. Preoperatively, plasma levels of both noradrenaline and adrenaline were significantly higher in patients with CHF compared with the control group. CABG was associated with clinical and haemodynamic improvement in all patients. Lymphocyte beta-adrenergic receptor density and function were also improved. Basal beta-adrenergic receptor density improved from 39.2 +/- 1.7 to 46 +/- 1.5 fmol/mg protein. Basal adenylyl cyclase activity increased from 33.1 +/- 2.6 40 +/- 2.4 pmol/mg protein per min; a significant increase in activity stimulated by isoproterenol (from 41.5 +/- 3.1 to 61 +/- 3.8 pmol/mg protein per min) and by NaF (from 71.8 +/- 2.7 to 85.3 +/- 3.5 pmol/mg protein per min) was also observed. Although postoperative plasma catecholamine levels tended to decrease, the difference compared with preoperative values was not significant. CONCLUSION: The results of our study indicate that CABG in patients with CHF is accompanied by the restoration of an almost normal functional state of the lymphocytic beta-adrenergic receptor system. PMID- 8723019 TI - Immunohistochemical characterization of immune cell composition and cytokine receptor expression in human coronary atherectomy tissue. AB - BACKGROUND: Prior histologic studies have examined smooth muscle cell, macrophage and thrombus constituents of atherosclerotic coronary atherectomy specimens. Lymphocytes and mononuclear leukocytes are also detectable in atherosclerotic surgical pathology specimens utilizing immunocytochemical techniques. METHODS: In order to quantify the histological contribution of cytokine receptor-expressing immunocompetent cells to human coronary artery stenoses, 30 directional atherectomy catheter biopsy specimens (wet weight < or = 10 mg) from 16 patients were snap frozen (-70 degrees C) for quantitative immunocytochemical studies. Following computer-assisted quantification of total intimal nuclei per tissue section (mean 297 +/- 177; cell density 7 +/- 5/10(4) microns 2), monoclonal antibody cytochemistry was used to identify the percentage of these cells expressing antigenic clusters of differentiation (CD) characteristic of T lymphocytes, B-lymphocytes and monocytes. Identification of alpha (low affinity) and beta (intermediate affinity) interleukin-2 receptors on intimal cells was accomplished using a three-step streptavidin-biotin method. RESULTS: A significant percentage of intimal cells were of lymphocytic (11 +/- 13%) or monocytic (12 +/- 14%) origin, with helper T-cells (9 +/- 12%) outnumbering both suppressor T-cells (2 +/- 4%) and B-lymphocytes (1 +/- 2%). Interleukin-2 receptors were noted on 9 +/- 12% of intimal cells, including cells with a vascular smooth muscle phenotype. CONCLUSIONS: These quantitative immunocytochemical data conclusively demonstrate that lymphocytes and monocytes account for over 20% of coronary plaque cells obtained by in-vivo atherectomy, and that helper (CD4) T-cells predominate over suppressor (CD8) T-cells and B lymphocytes. Variable interleukin-2 receptor subtype expression occurs in mononuclear leukocytes infiltrating chronic human atheroma. By applying these techniques, the therapeutic effects of cytotoxic agents on selectively targeted cytokine receptor-expressing cells may now be evaluated in vivo in small human directional coronary atherectomy specimens. PMID- 8723020 TI - Risk stratification for malignant arrhythmic events in patients with an acute myocardial infarction: role of an open infarct-related artery and the signal averaged ECG. AB - BACKGROUND: There is increasing evidence that an open infarct-related artery results in increased electrical stability of the heart and that this effect is at least in part responsible for the favorable outcome of these patients. In the thrombolytic era the optimal strategy for risk stratification remains controversial. This study examined the predictors of serious arrhythmic events during the first year after myocardial infarction. METHODS: A total of 222 patients with acute myocardial infarction, 41.4% of whom were treated with thrombolysis, were studied. At hospital discharge, signal-averaged electrocardiography was performed on 196 subjects and Holter monitoring on 200. One hundred and ninety-seven patients underwent coronary angiography. Left ventricular ejection fraction was determined in 201 subjects. RESULTS: An open infarct-related artery was documented in 106 patients. The incidence of late potentials was 34% (66 patients). Twenty-four patients (10.8%) had an arrhythmic event during follow-up (sudden death in seven, sustained ventricular tachyarrhythmias in 15, unexplained syncope in two). Signal-averaged electrocardiography had a sensitivity of 94% and a specificity of 72% for prediction of arrhythmic events. An occluded infarct-related artery was 78% sensitive and 58% specific, a left ventricular ejection fraction below 40% had a sensitivity of 71% and specificity of 80%, and Holter monitoring was only 38% sensitive and 92% specific. A combination of late potentials plus an occluded infarct-related artery was 68% sensitive and 84% specific. Positive predictive value was low for all variables examined, but could be improved by the combination of several risk factors. The highest positive predictive value was provided by the combination of an abnormal signal-averaged ECG and complex ventricular arrhythmias on ambulatory ECG. On multivariate analysis, in rank order, presence of late potentials, ejection fraction below 40%, high-grade ventricular ectopic activity and an occluded infarct-related artery were predictive of arrhythmic events. CONCLUSION: Among patients surviving an acute myocardial infarction, the occurrence of malignant arrhythmic events can be reliably predicted by the combination of an abnormal signal-averaged ECG, left ventricular dysfunction, complex ventricular arrhythmias on Holter monitoring and an occluded infarct-related artery at the time of hospital discharge. PMID- 8723022 TI - Bibliography of the current world literature. PMID- 8723021 TI - Alteration of lipid profile in hyperlipidemic rabbits by allicin, an active constituent of garlic. AB - BACKGROUND: The effect of garlic on the serum lipid profile has been the subject of controversy. This study was therefore designed to examine the effects of allicin, an active constituent of garlic, on the lipid profile in a rabbit model. METHODS: Allicin was produced by reacting alliin, synthesized in our laboratory, with purified alliinase. Nineteen New Zealand White rabbits were fed a cholesterol-rich diet (0.25% cholesterol) for 18 weeks. Ten rabbits received freshly produced allicin (3 mg/kg orally) starting at 8 weeks, and nine received placebo. There was no significant difference between the lipid profiles of the two groups at baseline up to 8 weeks. RESULTS: From day 28 of allicin supplementation a significant difference was found between the allicin and placebo groups in the graph regression lines describing the influence of allicin on serum cholesterol: Y = 41.39 + 8.69 multiplied by day (control) versus Y = 877.24 + 17.67 multiplied by day (allicin). The same trend was found for low density lipoprotein concentrations: Y = 10.3 + 8.4 multiplied by (control) versus Y = -750.4 + 15.7 multiplied by day (allicin). The serum high-density lipoprotein levels also differed significantly between the groups: Y = 20.29 + 0.24 multiplied by day (control) versus Y = -109.9 + 1.65 multiplied by day (allicin). CONCLUSIONS: Our results indicate that allicin has a beneficial effect on the serum lipid profile in hyperlipidemic rabbits, and should be further tested clinically. PMID- 8723023 TI - The angiotensin II subtype 2 (AT2) receptor is linked to protein kinase C but not cAMP-dependent pathways in the cardiomyocyte. AB - To define the signal transduction pathway linked to the cardiac angiotensin II (AII) receptor type 2 (AT2-R), cardiomyocytes were prepared as primary culture from 7-day-old chick embryo hearts. Protein kinase C (PKC) activity was assayed in membrane and cytosolic fractions of the cardiac cells. AII significantly (p < 0.05) increased membrane PKC activity and decreased cytosolic PKC activity, in a concentration-dependent manner, suggesting a translocation of PKC from cytosol to membrane. AT2-R blockade by its antagonist, PD123319, produced a dose-dependent antagonism of AII-induced activation of PKC. PD123319, at 10(-6) M or greater concentrations, completely antagonized AII-induced PKC activation, in contrast to a small reduction in PKC activity by the AII receptor type 1 (AT1-R) antagonist losartan. Isoproterenol-induced cAMP generation was significantly (p < 0.05) blunted by AII. AT2-R blockade did not alter AII-induced reduction of beta adrenoceptor-mediated increases in cAMP generation, in contrast with AT1-R blockade, which abolished AII-induced reduction of beta-adrenergic-mediated cAMP production. These data suggest that AT2 receptors have a previously unrecognized, role in the heart, namely, coupling AII to PKC. PMID- 8723024 TI - Effect of age on stretch-induced secretion of atrial natriuretic factor. AB - Plasma atrial natriuretic factor (ANF) levels are known to be higher in aged rats and humans. Although this may be partially explained by a reduction in clearance from the circulation, it was not known whether the secretory nature of the atrial tissue also changes with age. We measured ANF release in response to atrial distention in young adult (2-3 months) and older retired breeder (4-6 months) male rats both in vivo (conscious instrumented animals) and in vitro (isolated perfused atria). Whereas increased intraluminal pressure caused a rise in ANF secretion when using atria derived from the younger rats, there was no such response when using atria derived from the older rats, i.e., stretch induced secretion was impaired. This appeared to be secondary to a reduction in atrial compliance in the older animals. Unlike previous studies, basal plasma ANF levels were lower in our retired breeders, although the response to volume loading was preserved. These results suggest that, already by about 6 months of age, the atrial tissue is less responsive to changes in atrial pressure. The increased plasma levels previously reported in senescent rats are therefore probably a result of reduced clearance rather than increased secretion. PMID- 8723025 TI - In vitro and in vivo characterization of bradykinin B2 receptors in the rabbit and the guinea pig. AB - A comparative study has been performed in isolated organs and in anesthetized animals, rabbits, and guinea pigs, to evaluated the myotropic responses (in the organs) and the blood pressure changes (in the animals) induced by bradykinin (BK) and related peptides. Antagonist affinities have also been estimated in vitro in terms of PA2 and in vivo in terms of ID50, to characterize the kinin B2 receptors in the two species. Differences have been found both in the order of potency of agonists and in the affinity of antagonists: in fact, in the rabbit, [Hyp3]BK > [Aib7]BK, is the opposite order of what is found in the guinea pig, namely, [Aib7]BK < [Hyp3]BK, both in vitro and in vivo. Results obtained with antagonists also show important differences between the two species, since DArg[Hyp3, DPhe7, Leu8]BK is more active in the rabbit than in the guinea pig, while WIN-64338 is fairly active in the guinea pig and almost inactive in the rabbit. HOE-140, the long-acting antagonist of the B2 receptor, shows similar affinities in vitro in the two species. In another series of experiments, peptide degradation by angiotensin converting enzyme (ACE) has been investigated to see whether the differences of potency observed between certain peptides interacting with the B2 receptor were due to metabolic degradation. When incubated in the presence of pure ACE from rabbit lung, BK,[Hyp3]BK, and des Arg9BK are readily degraded, while [Aib7]BK, HOE-140, and DArg[Hyp3, DPhe7, Leu8]BK are not. When applied intravenously (i.v.), to obtain degradation by the lung, and intraarterially (i.a.), to avoid such degradation, the effect of BK (i.v.) is markedly reduced (compared with the effect i.a.), while no difference is observed for [Aib7]BK. Thus, despite its resistance to degradation by ACE, [Aib7]BK shows very little activity in the rabbit, suggesting that the major cause in the variation of affinities observed between kinin analogs is related to their pharmacodynamic properties. Taken together, the results speak strongly in favor of the existence of B2 receptor subtypes in the peripheral circulation of the rabbit and the guinea pig. Results obtained in vivo, both in pharmacological and biochemical experiments, are in accord with the findings obtained in isolated organs and with purified ACE enzyme. PMID- 8723026 TI - A method to evaluate the response of the coronary circulation of perfused rat heart to adenosine. AB - Exogenous adenosine causes a monophasic dilation of the coronary vessels in paced, perfused rat heart preparations. Because levels of endogenous adenosine in paced hearts may mask the presence of high potency adenosine receptors, we have developed a method to measure coronary vascular responses in a potassium-arrested heart. Hearts from adult male, Wistar rats were perfused at a constant flow rate of 10 mL/min in the nonrecirculating, Langendorff mode, using Krebs-Henseleit buffer. After 30 min, coronary perfusion pressure was 44 +/- 1 mmHg (mean +/- SEM). Hearts were then perfused with a modified Krebs-Henseleit buffer containing 35 mM potassium. Coronary perfusion pressure increased by 84 +/- 3 mmHg. Adenosine-induced reductions in coronary perfusion pressure were expressed as a percentage of the maximal increase in pressure produced by modified Krebs Henseleit buffer from the equilibration level. A concentration-response curve for adenosine (n = 6) was biphasic and best described by the presence of two adenosine receptors, with negative log EC50 values of 8.8 +/- 0.3 and 4.3 +/- 0.1, representing 29 +/- 3 and 71 +/- 3%, respectively, of the observed response. Interstitial adenosine sampled by microdialysis during potassium arrest was 25% of the concentration found in paced hearts. Endogenous adenosine in nonarrested hearts may obscure the biphasic response of the coronary vessels to adenosine. PMID- 8723027 TI - Modulation of the expression on constitutive rat hepatic cytochrome P450 isozymes by 5-fluorouracil. AB - The objective of this study was to determine the effects of 5-fluorouracil (5FU) on the expression of individual cytochrome P450 (CYP) isozymes in rat liver at the catalytic activity and apoprotein levels. Male Fischer 344 rats (9 to 10) weeks old) received a single intraperitoneal injection of 5FU (120 mg/kg) or vehicle. Rats were euthanized 1, 2, 7, or 14 days following drug administration. Hepatic microsomes were isolated and used for determination of spectral CYP and heme content and steroid hydroxylation activity, and immunoblot analysis of CYP apoproteins. 5FU treatment did not alter the levels of total microsomal CYP and heme. The major male-specific CYP isozyme, CYP2C11, was downregulated by 5FU treatment, as revealed by a significant decrease in CYP2C11 immunoreactive protein and catalytic activity (progesterone 2 alpha-hydroxylase) levels at day 14. Members of the CYP3A subfamily also appeared to be modulated by 5FU treatment in a complex manner. Two days following 5FU exposure, CYP3A immunoreactive protein was increased compared with vehicle control; however, 7 days after treatment, both CYP3A immunoreactivity and catalytic activity (progesterone 6 beta-hydroxylase) were suppressed by 5FU. 5FU appears to modulate the expression of constitutive CYP isozymes in the liver of male rats. The modulation of the catalytic activity of CYP2C11 and CYP3A by 5FU appears to be due to changes in the expression of the corresponding proteins. PMID- 8723028 TI - Effects of hemorrhagic hypotension on cerebral blood flow and perfused capillaries in newborn pigs. AB - We examined the effect of hemorrhagic hypotension on cerebral blood flow and perfused capillaries in newborn pigs, 2-10 days old. Cerebral blood flow was measured by using radioactive microspheres, perfused capillaries were determined by infusing a plasma marker, fluorescein isothiocyanate (FITC) - dextran (molecular mass 147,000 Da), and total capillaries were determined by staining with alkaline phosphatase. Rapid removal of 66 +/- 16 mL of blood resulted in a fall in mean blood pressure from 68 +/- 6 to 31 +/- 4 mmHg (1 mmHg = 133.3 Pa), an increase in heart rate from 137 +/- 18 to 240 +/- 34 beats/min, and a drop in arterial pH from 7.33 +/- 0.05 to 7.23 +/- 0.07. PCO2 was controlled by mechanical ventilation (36 +/- 4 mmHg before hemorrhage and 35 +/- 5 mmHg after hemorrhage) and PO2 remained stable (89 +/- 11 mmHg before hemorrhage and 94 +/- 10 mmHg after hemorrhage). Blood flow (n = 9) did not fall significantly in any brain region after hemorrhage. The percentage of perfused capillaries/mm2 (control, n = 7, hemorrhage, n = 6) was reduced in all brain regions during hypotension (cortex from 72 +/- 8 to 57 +/- 8%, cerebellum from 75 +/- 10 to 52 +/- 10%, and medulla from 76 +/- 8 to 51 +/- 9%). Similar results were seen for perfused capillary surface area We conclude that hemorrhagic hypotension resulted in a variable blood flow response with no significant reduction in cerebral blood flow but a reduction in perfused capillary number and surface area in all brain regions studied. We speculate that decreased capillary perfusion may be a contributing factor in diffuse neuronal injury after severe hemorrhagic hypotension. PMID- 8723029 TI - Prevention of diabetes in the spontaneously diabetic BB rat by the glutamine antimetabolite acivicin. AB - The autoimmune syndrome of the BB rat is associated with a marked increase in glutamine (Gln) metabolism in immune system cells of both diabetes-prone (BBdp) and diabetic (BBd) rats. To test whether inhibition of Gln metabolism prevents diabetes, 17 BBdp received acivicin (1 mg/kg) and 17 received saline subcutaneously every 2 days from age 48 days until diabetes onset or age 186 days. Twenty-seven non-diabetes-prone (BBn) rats served as controls. Acivicin caused some growth effects and a macrocytic anemia, but no other clinical or biochemical side effects. Only one acivicin-treated BBdp became diabetic (age 158 days), compared with saline-treated rats, of which 10 became diabetic and 2 became glucose intolerant (p < 0.001). Insulitis was moderate to severe in 88% of the saline-treated BBdp rats, but minimal in most acivicin-treated BBdp rats. Liver glutamine and glutamate tended to be higher in acivicin- than saline treated BBdp rats. Acivicin caused no change in the proportions of T or B lymphocytes, NK cells, or macrophage phenotypes in spleen or blood; all BBdp rats were typically lymphopenic. Mitogenic responses of splenocytes in vitro were not affected. The results are consistent with the hypothesis that acivicin, by interfering with Gln metabolism, "targets" activated cells of the immune system and thereby attenuates the process and prevents overt diabetes, without major disturbance of Gln levels or generalized immunosuppression. This prevention is not due to a nutritional-growth retardation effect, as diabetes was prevented in females that showed no such effect. PMID- 8723030 TI - Interactions between angiotensin II and adenosine 3':5'-cyclic monophosphate in the regulation of amino acid transport by vascular smooth muscle cells. AB - The regulation of amino acid transport by angiotensin II (AII) and cyclic AMP (cAMP) was assessed in cultured vascular smooth muscle cells, using a nonmetabolizable amino acid, alpha-[3H]aminoisobutyric acid (AIB). An exposure time in excess of 2 h was required for AII to elicit a stimulatory response, the magnitude of which increased in a time-dependent manner for 12 h. AII-induced transport was blocked by [1-sarcosine, 8-isoleucine]AII, a competitive inhibitor of AII binding. The effect of AII was not abolished by downregulation protein kinase C with phorbol 12,13-dibutyrate or by use of a protein kinase C inhibitor, suggesting that transport in response to AII can be mediated by a protein kinase C independent pathway. In contrast, the elimination of calcium from the incubation medium reduced AII-stimulated AIB uptake. The calmodulin inhibitor N (6-aminohexyl)-5-chloro-1-naphthalenesulfonamide partially inhibited AIB uptake in response to AII, suggesting that calmodulin may be involved in the modulation of AII-stimulated amino acid transport. AIB transport was also increased by elevating intracellular cAMP levels via beta-adrenergic receptor stimulation, the use of a cAMP analog (N6-monobutyryl cAMP), or a phosphodiesterase inhibitor (3 isobutyl-1-methylxanthine) or by direct stimulation of adenylate cyclase with forskolin. cAMP-induced AIB transport was evident within 10 min and peaked within 1 h. At 1 h AII enhanced cAMP-stimulated AIB transport. A possible mechanism for this effect is suggested by the observation that AII potentiated cAMP production in response to isoproterenol and 3-isobutyl-1-methylxanthine. PMID- 8723031 TI - Selective inhibition of oxalate-stimulated Ca2+ transport by cyclopiazonic acid and thapsigargin in smooth muscle microsomes. AB - 45Ca2+ uptake and efflux studies were performed on membranes prepared from dog mesenteric artery and rat vas deferens. Oxalate-stimulated, ATP-dependent Ca2+ uptake in microsomal vesicles, a property characteristic of sarcoplasmic reticulum, was completely inhibited in a concentration-dependent manner by cyclopiazonic acid (0.1-30 microM) and thapsigargin (10 nM-10 microM). Using discontinuous sucrose gradient centrifugation, rat vas deferens microsomes were separated into two fractions, one enriched in plasma membrane (F2), the other enriched in sarcoplasmic reticulum (F3). The F3 fraction had a major increase in Ca2+ uptake in the presence of oxalate, which was completely inhibited by either cyclopiazonic acid or thapsigargin. In the F2 fraction Ca2+ uptake in the presence of oxalate was lower than in F3 and was not completely inhibited by thapsigargin and cyclopiazonic acid. Instead, the F2 fraction had a thapsigargin insensitive and cyclopiazonic acid insensitive, saponin-sensitive component of uptake, which probably represents Ca2+ uptake by plasma membrane. In the absence of oxalate, the inhibition of Ca2+ uptake by saponin and cyclopiazonic acid or thapsigargin was additive in the F2 and F3 fractions, suggesting that cyclopiazonic acid and thapsigargin selectively inhibited sarcoplasmic reticulum derived Ca2+ uptake and did not affect plasma membrane derived Ca2+ uptake. Measurement of the initial rate of Ca2+ uptake in the presence and absence of oxalate by rat vas deferens microsomes demonstrated selective inhibition of oxalate-stimulated Ca2+ uptake by cyclopiazonic acid and thapsigargin. Ca2+ efflux from rat vas deferens microsomes actively loaded with 45Ca2+ either in presence or the absence of oxalate was not increased by cyclopiazonic acid or thapsigargin, showing that the inhibition of Ca2+ accumulation was not due to an increase in Ca2+ efflux. In both rat vas deferens and dog mesenteric artery, the maximal inhibitory effects of cyclopiazonic acid developed rapidly, whereas for maximal inhibition thapsigargin required pretreatment of microsomes prior to measurement of Ca2+ uptake. In rat vas deferens microsomes the inhibitory effects of cyclopiazonic acid could be quickly and completely reversed, whereas the effects of thapsigargin were not easily reversed. Collectively, these results suggest selectivity of cyclopiazonic acid and thapsigargin for the sarcoplasmic reticulum Ca2+ pump. Their selective inhibitory properties and differences in onset and offset of inhibition make cyclopiazonic acid thapsigargin useful pharmacological tools in the study of the physiological and pathophysiological roles of the sarcoplasmic reticulum Ca2+ pump in regulating smooth muscle Ca2+. PMID- 8723032 TI - The mode of rabbit platelet shape change and aggregation induced by theonezolide A, a novel polyketide macrolide, isolated from the Okinawan marine sponge Theonella sp. AB - Theonezolide-A (TZ-A), a novel polyketide macrolide, isolated from the Okinawan marine sponge Theonella sp., caused a marked platelet shape change at low concentrations (0.2-0.6 microM). Increasing concentrations of TZ-A to 6 microM or more caused shape change followed by a small but sustained aggregation. In a Ca(2+)-free solution, TZ-A-induced aggregation was markedly inhibited, although the marked shape change was still observed. Aggregation stimulated by TZ-A increased in a linear fashion with increasing Ca2+ concentrations from 0.1 to 3.0 mM. Furthermore TZ-A markedly enhanced 45Ca2+ uptake into platelets. Aggregation induced by TZ-A was inhibited by Arg-Gly-Asp-Ser, an inhibitor of fibrinogen binding to glycoprotein IIb-IIIa, H-7 and staurosporine, protein kinase C inhibitors, or genistein and tyrphostin A23, protein tyrosine kinase inhibitors, whereas shape change was blocked by genistein and tyrphostin A23. H-7 or staurosporine did not affect the TZ-A-induced shape change. These results suggest that TZ-A-induced platelet shape change is not dependent on external Ca2+, whereas TZ-A-induced aggregation is caused by an increase in Ca2+ permeability of the plasma membrane. It is also suggested that both aggregation and shape change induced by TZ-A are associated with protein phosphorylation by protein kinase C and tyrosine kinase. PMID- 8723033 TI - Molecular biomarkers for aflatoxins: from adducts to gene mutations to human liver cancer. AB - Over the past 30 years there have been extensive efforts to investigate the association between aflatoxin exposure and human liver cancer. These studies have been hindered by the lack of adequate dosimetry data on aflatoxin intake, excretion, and metabolism in people, as well as by the general poor quality of worldwide cancer morbidity and mortality statistics. These realities have spurred the efforts to develop new technologies to assess exposure status and risk for aflatoxins, and these agents are among the few environmental carcinogens for which quantitative risk assessments have been attempted. One of the goals of these risk assessments has been the development of primary and secondary preventive intervention methods to lower the human health impact from aflatoxin exposures. The long-term goal of the research described herein is the application of biomarkers to the development of preventive interventions for use in human populations at high risk for cancer. Several of the aflatoxin-specific biomarkers have been validated in epidemiologic studies and are now available for use as intermediate biomarkers in prevention trials. The development of these aflatoxin biomarkers has been based upon the knowledge of the biochemistry and toxicology of aflatoxins gleaned from both experimental and human studies. These biomarkers have been utilized subsequently in experimental models to provide data on the modulation of the markers under different situations of disease risk. This systematic approach provides encouragement for preventive interventions and should serve as a template for the development for the development and validation of other chemical-specific biomarkers and their application to cancer or other chronic diseases. PMID- 8723034 TI - Altered porphyrin metabolism as a biomarker of mercury exposure and toxicity. AB - Changes in urinary porphyrin excretion patterns (porphyrin profiles) have been described in response to a variety of drugs and chemicals. The present studies were conducted to define the specific changes in the urinary porphyrin profile associated with prolonged exposure to mercury and mercury compounds. In rats, exposure for a prolonged period to mercury as methyl mercury hydroxide was associated with urinary porphyrin changes, which were uniquely characterized by highly elevated levels of 4- and 5-carboxyl porphyrins and by the expression of an atypical porphyrin ("precoproporphyrin") not found in urine of unexposed animals. These distinct changes in urinary porphyrin concentrations were observed as early as 1-2 weeks after initiation of mercury exposure, and increased in a dose- and time-related fashion with the concentration of mercury in the kidney, a principal target organ of mercury compounds. Following cessation of mercury exposure, urinary porphyrin concentrations reverted to normal levels, consistent with renal mercury clearance. In human studies, a comparable change in the urinary porphyrin profile was observed among subjects with occupational exposure to mercury as mercury vapor sufficient to elicit urinary mercury levels greater than 20 micrograms/L. Urinary porphyrin profiles were also shown to correlate significantly with mercury body burden and with specific neurobehavioral deficits associated with low level mercury exposure. These findings support the utility of urinary porphyrin profiles as a useful biomarker of mercury exposure and potential health effects in human subjects. PMID- 8723035 TI - Assessing the estrogenic and dioxin-like activities of chemicals and complex mixtures using in vitro recombinant receptor-reporter gene assays. AB - In vitro recombinant receptor-reporter gene assays have been used to assess and rank the potency of chemicals and complex mixtures suspected of possessing estrogen and (or) aryl hydrocarbon receptor (AhR) mediated activity. The environmental estrogen (E2) bioassay consists of a Gal4-human estrogen receptor chimeric construct (Gal4-HEGO) and a Gal4-regulated luciferase reporter gene (17m5-G-Luc) that have been stably integrated into HeLa cells. The assay exhibits 10-fold induction in luciferase reporter gene activity following treatment with 1 nM 17 beta-estradiol and has a detection limit of approximately 5 pg of 17 beta estradiol/mL. The AhR bioassay uses Hepa 1c1c7 wild-type cells transiently transfected with a dioxin response element regulated luciferase reporter gene. These assays were used to assess the estrogen and dioxin-like activities of naringenin, atrazine, and simazine and complex mixtures such as pulp and paper mill black liquor and urban air particulates. The activities of these chemicals and complex mixtures are confirmed using the pure antiestrogen ICI 164,384 and in in vitro gel retardation assays. Results of this study demonstrate the utility of in vitro recombinant receptor-reporter gene assays in identifying and assessing the estrogenic and dioxin-like activities of chemicals and complex mixtures. PMID- 8723036 TI - Linkage results in schizophrenia. PMID- 8723037 TI - Reflections on the evidence for a vulnerability locus for schizophrenia on chromosome 6p24-22. PMID- 8723038 TI - Neurobehavioral profiles of children with neurofibromatosis 1 referred for learning disabilities are sex-specific. AB - We compared neurobehavioral profiles of 10 children with neurofibromatosis 1 (NF 1) referred for evaluation of learning disabilities (NF/LD) to those to learning disabled children without known genetic disease (LD), matched for age, sex, and estimated IQ. It was hypothesized that the NF/LD children would exhibit a neurobehavioral profile diagnostic of compromise of frontal/subcortical brain systems while those of the case controls would be heterogeneous. Records from a clinical data base were reviewed retrospectively for the neurological and neuropsychological components of an interdisciplinary learning disabilities evaluation. Neurological abnormalities were more frequent in the NF/LD group, involving gross and fine motor coordination, praxis, and megencephaly. As predicted, clinical neuropsychological diagnostic ratings and composite neurobehavioral observation scores were consistent with compromise of frontal systems in the NF/LD group. An unanticipated finding was that outcomes in the NF/LD group were sex dependent: Megencephaly was observed in females only; and the frontal/subcortical neurobehavioral profile was more consistently observed in females. Females with NF-1 with megencephaly may be at increased risk for a neurobehavioral syndrome contributing to LD that is consistent with compromise of frontal/subcortical brain systems. PMID- 8723039 TI - Further association study on dopamine D2 receptor variant S311C in schizophrenia and affective disorders. AB - The dopamine D2 receptor gene is a candidate gene for schizophrenia because the potency of certain neuroleptics correlates with their affinity for this receptor. Case-control studies in 291 schizophrenics, 78 patients with affective disorders, and 579 controls on an association of a molecular variant of S311C of the dopamine D2 receptor with psychiatric disorders were conducted. The frequency of individuals with S311C was significantly higher in schizophrenics with the absence of negative symptoms (17.1%, P < 0.00001), but similar in schizophrenics with the presence of negative symptoms (5.7%, P = 0.46) when compared with the controls (4.1%). The frequency of S311C was significantly higher in familial schizophrenics from one local area but not in those from other areas. It was significant that S311C was frequently present in patients with mood-incongruent psychotic affective disorders (33.3%, P < 0.0001), but not in those with other affective disorders. These data suggest that S311C might be one of the genetic factors for symptomatic dimensions of delusions and hallucinations and might be involved in underlying clinical heterogeneity in schizophrenia and affective disorders. PMID- 8723041 TI - New segregation analysis of panic disorder. AB - We performed simple segregation analyses of panic disorder using 126 families of probands with DSM-III-R panic disorder who were ascertained for a family study of anxiety disorders at an anxiety disorders research clinic. We present parameter estimates for dominant, recessive, and arbitrary single major locus models without sex effects, as well as for a nongenetic transmission model, and compare these models to each other and to models obtained by other investigators. We rejected the nongenetic transmission model when comparing it to the recessive model. Consistent with some previous reports, we find comparable support for dominant and recessive models, and in both cases estimate nonzero phenocopy rates. The effect of restricting the analysis to families of probands without any lifetime history of comorbid major depression (MDD) was also examined. No notable differences in parameter estimates were found in that subsample, although the power of that analysis was low. Consistency between the findings in our sample and in another independently collected sample suggests the possibility of pooling such samples in the future in order to achieve the necessary power for more complex analyses. PMID- 8723040 TI - Analysis of thirteen trinucleotide repeat loci as candidate genes for schizophrenia and bipolar affective disorder. AB - A group of diseases are due to abnormal expansions of trinucleotide repeats. These diseases all affect the nervous system. In addition, they manifest the phenomenon of anticipation, in which the disease tends to present at an earlier age or with greater severity in successive generations. Many additional genes with trinucleotide repeats are believed to be expressed in the human brain. As anticipation has been reported in schizophrenia and bipolar affective disorder, we have examined allele distributions of 13 trinucleotide repeat-containing genes, many novel and all expressed in the brain, in genomic DNA from schizophrenic (n = 20-97) and bipolar affective disorder patients (23-30) and controls (n = 43-146). No evidence was obtained to implicate expanded alleles in these 13 genes as causal factors in these diseases. PMID- 8723042 TI - Psychiatric disorder in a familial 15;18 translocation and sublocalization of myelin basic protein of 18q22.3. AB - Two related patients with similar clinical features consisting of a few dysmorphic signs and psychiatric disturbance were reported to have a partial trisomy of chromosomes 15(pter-q13.3) and 18(q23-qter) deriving from a familial translocation t(15;18). One patient is affected by bipolar disorder and the other by schizoaffective disorder. Both cases have a predominantly affective course; nevertheless, a clear diagnosis is difficult in the first patient, who is 15 years of age, and only a longitudinal course will allow us to establish a definite diagnosis. The possibility that these two pathologies belong to a single category is discussed, and the presence of a susceptibility locus on chromosome 18 is hypothesized. Cytogenetic data, FISH, and DNA studies indicate that the myelin basic protein (MPB) gene is not involved in the translocation, and localize it centromeric to the breakpoint on chromosome 18(q22.3). Thus, it is unlikely to be involved in the disease. PMID- 8723043 TI - Evidence of a locus for schizophrenia and related disorders on the short arm of chromosome 5 in a large pedigree. AB - We attempted to identify a locus for schizophrenia and related disorders in 24 nuclear families of schizophrenic probands using a predefined classification system for affected cases that included those disorders most clearly identified as sharing a genetic relationship with schizophrenia--schizoaffective disorder and schizotypal personality disorder. Initially, we evaluated 8 markers on chromosome 5 on the first 12 families with available genotyping and diagnostic assessments and, assuming autosomal dominant transmission, found a lod score of 2.67 for the D5S111 locus (5p14.1-13.1) in one large nuclear family (no. 17; sibship: n = 12; schizophrenia: n = 3; schizotypal personality disorder: n = 2); the other 11 families were much smaller, less complete, and provided little additional information. Other branches of no. 17 were then assessed and the 2 point lod score for family 17 rose to 3.72; using multipoint analysis the lod score in 17 was 4.37. When only schizophrenia was used to define affectedness, the positive evidence for linkage to D5S111 was greatly reduced. Sensitivity analysis indicated that the lod score is heavily dependent upon the predefined diagnostic criteria. Our studies of other families of schizophrenic probands eventually totalled 23, but linkage to D5S111 in these yielded a -2.41 lod score. The results provide evidence for genetic linkage of the D5S111 locus to schizophrenia and related disorders in one family. It may be of interest that over several generations, almost all the ancestors of family 17 could be traced back to a small, relatively isolated, hill region of Puerto Rico. PMID- 8723044 TI - Neuropsychiatry of 18q- syndrome. AB - Our understanding of neuropsychiatric abnormalities in patients with deletions of the long arm of chromosome 18 (18q- syndrome) is based mainly on sporadic case reports. We characterized the neuropsychiatric phenotype in 27 patients across a wide age range (2-47 years) with breakpoints ranging from 18q22.3-18q21.2. Adaptive behavior scores (Vineland Composite) were significantly higher in females than in males (62 +/- 5 vs. 43 +/- 3). Intelligence ranged from borderline to severely deficient (IQ, 73- < 40), with academic achievement similarly impaired. Performance in specific neuropsychological functions, including attention, novel problem solving, memory, language, visuomotor integration, and fine motor dexterity, was consistently in the moderately-to severely impaired range. Behavioral problems were common in both sexes, including aggressivity, hyperactivity, and temper tantrums. Contrary to the few previous reports, we found no evidence of psychosis in any patients. In a subset of patients selected on the basis of no prior knowledge of behavioral problems, 1 of 16 patients (6%) had autism, as defined by the Autistic Diagnostic Interview- Revised (ADI-R) [Lord et al., 1994: J Autism Dev Disord 24:659-685]. Thus, the prevalence of autism in 18q- syndrome is probably no greater than that in other developmental disabilities with a similar level of cognitive impairment. In contrast to what has been believed since 18q- was first described 30 years ago, we found no relationship between chromosome deletion size and any measure of cognition or behavior; nor were there any correlations between any of these measures with the presence or absence of abnormalities on MRI or somatosensory evoked potentials. PMID- 8723045 TI - Irish study on high-density schizophrenia families: field methods and power to detect linkage. AB - Large samples of multiplex pedigrees will probably be needed to detect susceptibility loci for schizophrenia by linkage analysis. Standardized ascertainment of such pedigrees from culturally and ethnically homogeneous populations may improve the probability of detection and replication of linkage. The Irish Study of High-Density Schizophrenia Families (ISHDSF) was formed from standardized ascertainment of multiplex schizophrenia families in 39 psychiatric facilities covering over 90% of the population in Ireland and Northern Ireland. We here describe a phenotypic sample and a subset thereof, the linkage sample. Individuals were included in the phenotypic sample if adequate diagnostic information, based on personal interview and/or hospital record, was available. Only individuals with available DNA were included in the linkage sample. Inclusion of a pedigree into the phenotypic sample required at least two first, second, or third degree relatives with non-affective psychosis (NAP), one whom had schizophrenia (S) or poor-outcome schizo-affective disorder (PO-SAD). Entry into the linkage sample required DNA samples on at least two individuals with NAP, of whom at least one had S or PO-SAD. Affection was defined by narrow, intermediate, and broad criteria. The phenotypic sample contained 277 pedigrees and 1,770 individuals and the linkage sample 265 pedigrees and 1,408 individuals. Using the intermediate definition of affection, the phenotypic sample contained 837 affected individuals and 526 affected sibling pairs. Parallel figures for the linkage sample were 700 and 420. Individuals with schizophrenia from these multiplex pedigrees resembled epidemiologically sampled cases with respect to age at onset, gender distribution, and most clinical symptoms, although they were more thought-disordered and had a poorer outcome. Power analyses based on the model of linkage heterogeneity indicated that the ISHDSF should be able to detect a major locus that influences susceptibility to schizophrenia in as few as 20% of families. Compared to first-degree relatives of epidemiologically sampled schizophrenic probands, first-degree relatives of schizophrenic members from the ISHDSF had a similar risk for schizotypal personality disorder, affective illness, alcoholism, and anxiety disorder. With sufficient resources, large-scale ascertainment of multiplex schizophrenia pedigrees is feasible, especially in countries with catchmented psychiatric care and stable populations. Although somewhat more severely ill, schizophrenic members of such pedigrees appear to clinically resemble typical schizophrenic patients. Our ascertainment process for multiplex schizophrenia families did not select for excess familial risk for affective illness or alcoholism. With its large sample ascertained in a standardized manner from a relatively homogeneous population, the ISHDSF provides considerable power to detect susceptibility loci for schizophrenia. PMID- 8723046 TI - Unique case of growth hormone (GH) deficiency accompanied by clinical anophthalmia, hypoplastic orbits, digital dysplasia, short stature, obesity, and diabetes mellitus. AB - A 43-year-old female was admitted to our hospital for polydipsia and hyperglycemia. She had total blindness and globes were not recognized by inspection, indicating clinical anophthalmia. Physical examination revealed short stature, obesity, prematurely gray hair, shortness of fingers and toes, syndactyly, and multiple dental caries. Laboratory examination showed hyperglycemia, increased glycosilated hemoglobin (HbA1c) and insulin resistance on euglycemic glucose clamp. Blunted growth hormone (GH) secretion was shown in response to insulin-induced hypoglycemia, arginine infusion, and GH-releasing hormone (GHRH) loading test, and in 24 h spontaneous GH profile. Magnetic resonance imaging (MRI) and computed tomography (CT) showed dysostosis of orbit, defect of optic nerve, enlarged suprasellar cistern, and prolonged pituitary stalk. This may be the first report of a unique case with GH deficiency accompanied by clinical anophthalmia, hypoplastic orbits, digital dysplasia, short stature, obesity, and diabetes mellitus. PMID- 8723047 TI - Familial alcoholism in manic-depressive (bipolar) disease. AB - A previous analysis found a relatively high rate of alcoholism in a cohort of bipolar I subjects, and a trend for increased rates of alcoholism in relatives of subjects with both bipolar I disorder and alcoholism, compared to relatives of subjects with bipolar I disorder and no alcoholism. The sample of subjects with bipolar I disorder has been enlarged through continued follow-up, permitting new analyses to address the association and heritability of bipolar I disorder with alcoholism. Probands with bipolar I disorder were followed for 10 years as part of the NIMH Collaborative Depression Study. The rate of alcoholism in relatives of probands with both bipolar I disorder and alcoholism was compared to the rate of alcoholism in relatives of probands with bipolar disorder and no alcoholism. The prevalence of alcoholism in relatives of subjects with bipolar I disorder was compared to the rate of alcoholism in relatives of control subjects. Relatives of probands with bipolar I disorder showed a higher rate of alcoholism than relatives of controls. Relatives of probands with bipolar I disorder and alcoholism showed a higher rate of alcoholism than relatives of probands with bipolar I disorder without alcoholism. These data suggest that familial alcoholism may contribute to a vulnerability to bipolar I disorder, and that there is a shared heritability for the two disorders. PMID- 8723048 TI - Maternal inheritance and chromosome 18 allele sharing in unilineal bipolar illness pedigrees. AB - We have replicated the observation of McMahon et al. [1995] that there is excess maternal transmission of illness in a series of previously described unilineal Bipolar manic-depressive illness extended pedigrees [Berrettini et al., 1991]. ("Transmission" is defined for any ill person in a pedigree when father or mother has a personal or immediate family history of major affective disorder.) We divided our pedigrees into exclusively maternal transmission (Mat) and mixed maternal-paternal transmission (in different pedigree branches) (Pat). Using affected sib-pair-analysis, linkage to a series of markers on chromosome 18p-cen was observed in the Pat but not the Mat pedigrees, with significantly greater identity by descent (IBD) at these markers in the Pat pedigrees. As compared with the pedigree series as a whole, the proportion of alleles IBD in the linkage region is much increased in the Pat pedigrees. As shown by Kruglyak and Lander [1995], as the sharing proportion of alleles in affected relative pairs increases, the number of such pairs needed to resolve the linkage region to a 1 cM interval becomes smaller. Genetic subdivision of an illness by clinical or pedigree configuration criteria may thus play an important role in discovery of disease susceptibility mutations. PMID- 8723049 TI - Lack of association between dopamine D2 receptor gene Cys311 variant and schizophrenia. AB - Itokawa et al. [1993] reported identifying one missense nucleotide mutation from C to G resulting in a substitution of serine with cysteine at codon 311 in the third intracellular loop of the dopamine D2 receptor in schizophrenics. Arinami et al. [1994] reported finding a positive association between the Cys311 variant and schizophrenia. In response to the report by Arinami et al. [1994] we examined 106 unrelated Japanese schizophrenics and 106 normal controls to determine if there is any association of the Cys311 variant with schizophrenia. However, we found no statistically significant differences in allelic frequencies of Cys311 between schizophrenia and normal controls. The present results as well as those of all previous studies except for that of Arinami et al. [1994] indicated that an association between the dopamine D2 receptor gene and schizophrenia is unlikely to exist. PMID- 8723050 TI - Lujan-Fryns syndrome in the differential diagnosis of schizophrenia. AB - Schizophrenia is considered to be a heterogenous disorder. Different etiopathological mechanism can be attributed to a similar clinical picture as described in DSM-III-R criteria. We present a case of a young man diagnosed on different occasions as schizophrenic with mild mental retardation. Clinical examination revealed signs and symptoms most compatible with the diagnosis of Lujan-Fryns syndrome, an X-linked mental retardation syndrome with marfanoid features, frequently associated with psychotic or other psychiatric symptoms. In all patients with symptoms of schizophrenia and mental retardation Lujan-Fryns syndrome should be considered in the differential diagnosis. PMID- 8723052 TI - Linkage study between manic-depressive illness and chromosome 21. AB - Chromosome 21, of interest as potentially containing a disease gene for manic depressive illness as possible evidence for a gene pre-disposing to affective disorder, has recently been reported in a single large family as well as samples of families. The present study investigates for linkage between manic-depressive illness and markers covering the long arm of chromosome 21 in two manic depressive families, using ten microsatellite polymorphisms as markers. No conclusive evidence for a disease gene on the long arm of chromosome 21 was found. Assuming either a dominant or recessive mode of inheritance, close linkage to the marker PFKL, which has been reported as possibly linked to affective disorder, seems unlikely in the families studied here. PFKL and more telomeric markers yielded small positive lod scores at higher recombination fractions in the largest family, and small positive lod scores at lower recombination fractions in the affected-only analyses in the smallest family. PMID- 8723051 TI - Genetic linkage study of bipolar disorder and the serotonin transporter. AB - The serotonin transporter (HTT) is an important candidate gene for the genetic transmission of bipolar disorder. It is the site of action of many antidepressants, and plays a key role in the regulation of serotonin neurotransmission. Many studies of affectively ill patients have found abnormalities in serotonin metabolism, and dysregulation of the transporter itself. The human serotonin transporter has been recently cloned and mapped to chromosome 17. We have identified a PstI RFLP at the HTT locus, and here report our examination of this polymorphism for possible linkage to bipolar disorder. Eighteen families were examined from three populations: the Old Order Amish, Iceland, and the general North American population. In addition to HTT, three other microsatellite markers were examined, which span an interval known to contain HTT. Linkage analyses were conducted under both dominant and recessive models, as well as both narrow (bipolar only) and broad (bipolar + recurrent unipolar) diagnostic models. Linkage could be excluded to HTT under all models examined. Linkage to the interval spanned by the microsatellites was similarly excluded under the dominant models. In two individual families, maximum lod scores of 1.02 and 0.84 were obtained at D17S798 and HTT, respectively. However, these data overall do not support the presence of a susceptibility locus for bipolar disorder near the serotonin transporter. PMID- 8723054 TI - Lack of imprinting of the human dopamine D4 receptor (DRD4) gene. AB - The term genomic imprinting has been used to refer to the differential expression of genetic material depending on whether it has come from the male or female parent. In humans, the chromosomal region 11p15.5 has been shown to contain 2 imprinted genes (H19 and IGF2). The gene for the dopamine D4 receptor (DRD4), which is of great interest for research into neuropsychiatric disorders and psychopharmacology, is also located in this area. In the present study, we have examined the imprinting status of the DRD4 gene in brain tissue of an epileptic patient who was heterozygous for a 12 bp repeat polymorphism in exon 1 of the DRD4 gene. We show that both alleles are expressed in equivalent amounts. We therefore conclude that the DRD4 gene is not imprinted in the human brain. PMID- 8723053 TI - Systematic screening for mutations in the human serotonin 1F receptor gene in patients with bipolar affective disorder and schizophrenia. AB - Using single strand conformational analysis we screened the complete coding sequence of the serotonin 1F (5-HT1F) receptor gene for the presence of DNA sequence variation in a sample of 137 unrelated individuals including 45 schizophrenic patients, 46 bipolar patients, as well as 46 healthy controls. We detected only three rare sequence variants which are characterized by single base pair substitutions, namely a silent T-->A transversion in the third position of codon 261 (encoding isoleucine), a silent C-->T transition in the third position of codon 176 (encoding histidine), and an C-->T transition in position -78 upstream from the start codon. The lack of significant mutations in patients suffering from schizophrenia and bipolar affective disorder indicates that the 5 HT1F receptor is not commonly involved in the etiology of these diseases. PMID- 8723055 TI - No association detected between a D3 receptor gene-expressed variant and schizophrenia. AB - A missense polymorphism (glycine to serine) in the first exon of the dopamine D3 (DRD3) gene was examined in the sib-pairs schizophrenia collection by the transmission test for linkage disequilibrium (TDT). No association due to linkage disequilibrium was detected using TDT. Additionally, no evidence for excess homozygosity was found. PMID- 8723057 TI - Lack association between schizophrenia and the CYP2D6 gene polymorphisms. PMID- 8723056 TI - Apolipoprotein E genotype in schizophrenia. PMID- 8723058 TI - In vivo study of the mutagenicity of biperidine, pipotiazine, chlorpromazine, and haloperidol. PMID- 8723060 TI - Familial congenital bicuspid aortic valve: a disorder of uncertain inheritance. AB - Congenital bicuspid aortic valve (BAV) is one of the most frequent heart defects detected by echocardiographic investigation and necropsy (0.9-1% of the general population), but only 16 families with familial congenital BAV have been described up to now. We report on a family in which 4 members of two generations (2 brothers, 1 sister, and her son) are affected with BAV. The BAV mode of inheritance is discussed. PMID- 8723059 TI - Mosaic vs. nonmosaic trisomy 9: report of a liveborn infant evaluated by fluorescence in situ hybridization and review of the literature. AB - We report on a newborn infant with multiple congenital anomalies and apparent nonmosaic trisomy 9 in the blood (by conventional cytogenetic studies) who died shortly after birth. Clinical observations at birth and autopsy are compared with phenotypes of mosaic and nonmosaic trisomy 9 cases reported previously. Unlike the initial cytogenetic analysis, fluorescence in situ hybridization (FISH) studies of metaphase and interphase blood cells and skin fibroblasts detected the presence of euploid and trisomy 9 cells. These results suggest that earlier reports of trisomy 9, which relied on conventional chromosome analysis of a few metaphase cells and/or only one tissue type, may not have excluded mosaicism, and that trisomy 9 may be viable only in the mosaic state. PMID- 8723061 TI - Suture formation, premature sutural fusion, and suture default zones in Apert syndrome. AB - On the basis of our studies, we postulate that suture formation in Apert syndrome is related to the relative maturity of abutting calvarial bones. The fused coronal suture, a consistent manifestation at birth, develops first because the ossification centers of the frontal and parietal bones are in intimate contact early during intrauterine life. Calvarial immaturity and the megalencephalic brain characteristic of the Apert syndrome appear to work in concert to produce a widely patent midline calvarial defect extending from the glabella to the posterior fontanelle. Because sagittal growth in the coronal sutures cannot take place, the megalencephalic brain grows upward and laterally, and bulges forward through the midline defect. The defect fills in by coalescence of bony islands without proper suture formation because the gap to be bridged is so great that the time window for developing sutural interdigitations may have closed. Other sutures, such as the lambdoid, squamosal, and sphenotemporal, develop with normal interdigitations because abutting bone margins are in close enough proximity to permit suture formation. PMID- 8723062 TI - Interstitial deletion (6)q13q15. AB - We report on a 2-year-old child with psychomotor retardation, facial and urogenital anomalies. His chromosome constitution was 46,XY, del(6)(q13q15). This case further contributes to the karyotype-phenotype correlation of proximal deletion 6q syndromes. PMID- 8723063 TI - Linear disruption of umbilical cord: a rare anomaly of the cord associated with acute fetal distress and perinatal death/profound psychomotor retardation. AB - We report on a non-malformed child with severe microcephaly and profound psychomotor delay. Review of the delivery/birth records documented descriptions consistent with linear disruption of the umbilical cord. This rare anomaly typically leads to acute fetal distress and perinatal death. Severe microcephaly and psychomotor delay without other anomalies should prompt a careful review of the delivery/birth records to search for umbilical cord descriptions consistent with this diagnosis. PMID- 8723065 TI - Hereditary nonpolyposis colorectal cancer: review of clinical, molecular genetics, and counseling aspects. AB - Lynch syndrome, or hereditary nonpolyposis colon cancer (HNPCC), is an autosomal dominant disease accounting for approximately 1-5% of all colorectal cancer cases. Due to the lack of pathognomonic morphological or biomolecular markers, HNPCC has traditionally posed unique problems to clinicians and geneticists alike, both in terms of diagnosis and clinical management. Recently, novel insight into the pathogenesis of this syndrome has been provided by the identification of its molecular basis. In HNPCC families, germline mutations in any of four genes encoding proteins of a specialized DNA repair system, the mismatch repair, predispose to cancer development. Mutations in mismatch repair genes lead to an overall increase of the mutation rate and are associated with a phenotype of length instability of microsatellite loci. The present report summarizes the clinicopathological aspects of HNPCC and reviews the most recent molecular and biochemical findings. PMID- 8723064 TI - Deletion of small nuclear ribonucleoprotein polypeptide N (SNRPN) in Prader-Willi syndrome detected by fluorescence in situ hybridization: two sibs with the typical phenotype without a cytogenetic deletion in chromosome 15q. AB - The small nuclear ribonucleoprotein polypeptide N (SNRPN) gene is regarded as one of the candidates for Prader-Willi syndrome (PWS). We describe two sibs with typical PWS presenting deletion of SNRPN detected by fluorescence in situ hybridization (FISH). Neither a cytogenetically detectable 15q12 deletion nor a deletion for the D15S11, D15S10, and GABRB3 cosmid probes were found in either patient. This implies a smaller deletion limited to the PWS critical region. FISH with a SNRPN probe will permit analysis of PWS patients with limited deletions not detectable with other probes. PMID- 8723066 TI - Autosomal recessive disorder with muscle contractions resembling neonatal tetanus, characteristic face, camptodactyly, hyperthermia, and sudden death: a new syndrome? AB - This work describes an autosomal recessive syndrome observed over the past 25 years in 17 newborn babies (8 males, 9 females), from 12 different families in Southern Sardinia. This disorder is evident at birth and is characterized by marked muscular contraction of the facial muscles in response to tactile stimuli or during crying, with trismus and abundant salivation simulating a tetanic spasm. The contractions slowly disappear as the infant calms. There is also neck muscle hypertonia with a tendency to opisthotonus. All patients present facial anomalies such as large face, chubby cheeks, broad nose with anteverted nostrils, and long philtrum. The hands show bilateral camptodactyly. The clinical course in all patients was characterized by marked feeding difficulties and appearance of variable fever at about 38 degrees C, with peaks of irregular hyperthermia of over 42 degrees C, with onset ranging from birth to a few weeks. In some patients these symptoms were accompanied by generalized seizures. Death occurred after a period of a few weeks to some months and coincided with fever above 42 degrees C. Laboratory investigations performed in all of these cases did not give any useful pathogenetic indications. Only patients 10 and 16 are still alive today. Patient 10 is now 14 years old. She presents slow regression of the dystonic symptomatology, while dysthermia and mild psychomotor delay persist. PMID- 8723067 TI - Hunter-McAlpine craniosynostosis phenotype associated with skeletal anomalies and interstitial deletion of chromosome 17q. AB - Hunter-McAlpine syndrome is an autosomal dominant disorder consisting of variable manifestations including craniosynostosis, almond-shaped palpebral fissures, small mouth, mild acral-skeletal anomalies, short stature, and mental deficiency. We report on a 9-year-old boy with this phenotype with more severe skeletal abnormalities than previously described. Chromosomes showed del(17)(q23.1- >q24.2); the more severe phenotype may be explained by the deletion. The deletion also suggests the possibility that the gene for Hunter-McAlpine syndrome might map to that region. PMID- 8723068 TI - Rates of Down syndrome at livebirth by one-year maternal age intervals in studies with apparent close to complete ascertainment in populations of European origin: a proposed revised rate schedule for use in genetic and prenatal screening. AB - Precision and accuracy in determining rates of Down syndrome at livebirth are indispensible to algorithms which determine eligibility for prenatal cytogenetic diagnostic services. We derived Down syndrome rates by single year of maternal age which we propose as a revised rate schedule for background risk. Data on European-origin populations were obtained from 5 sources judged most likely to have complete ascertainment of cases. A "constant plus exponent" regression model and variants extending the analysis to higher powers of maternal age were applied to several ranges of maternal age. Confidence intervals about the rates were calculated. This analysis results in rates significantly higher than those in widespread use though the confidence intervals show a need for caution in assuming precision. Sources of variation in rates are also considered. PMID- 8723069 TI - Hypertrichosis, pigmentary retinopathy, and facial anomalies: a new syndrome? AB - We report on a 22-month-old male with congenital hypertrichosis of the face, arms, legs, shoulders, back, and buttocks, abnormal facial appearance, dolichocephaly, and pigmentary retinopathy. Symmetrical hyperpigmentation is present on the sideburn areas of his face, and hyperpigmented streaks are seen on arms and legs. Biopsy of the hyperpigmented' skin showed many separate bundles of smooth muscles in the dermis. No relative had hypertrichosis or other birth defects. To our knowledge, the syndrome of facial anomalies, pigmentary retinopathy, and congenital hypertrichosis has not been reported previously. PMID- 8723070 TI - Anophthalmia-Waardenburg syndrome: a report of three cases. AB - We report on 2 Turkish families with children who had bilateral anophthalmia, upper and lower limb abnormalities, mental retardation and consanguineous parents. We have evaluated the 2 cases in the first family and the only case in the second as anophthalmia-Waardenburg syndrome. This is an extremely rare autosomal recessive syndrome. PMID- 8723072 TI - The expanding clinical phenotype of the tRNA(Leu(UUR)) A-->G mutation at np 3243 of mitochondrial DNA: diabetic embryopathy associated with mitochondrial cytopathy. AB - We describe a family which demonstrates and expands the extreme clinical variability now known to be associated with the A-->G transition at nucleotide position 3243 of the mitochondrial DNA. The propositus presented at birth with clinical manifestations consistent with diabetic embryopathy including anal atresia, caudal dysgenesis, and multicystic dysplastic kidneys. His co-twin was normal at birth, but at 3 months of life, presented with intractable seizures later associated with developmental delay. The twins' mother developed diabetes mellitus type I at the age of 20 years and gastrointestinal problems at 22 years. Since age 19 years, the maternal aunt has had recurrent strokes, seizures, mental deterioration and deafness, later diagnosed as MELAS syndrome due to the tRNA(Leu(UUR)) A-->G mutation. A maternal uncle had diabetes mellitus type I, deafness, and normal intellect, and died at 35 years after recurrent strokes. This pedigree expands the known clinical phenotype associated with tRNA(Leu(UUR)) A-->G mutation and raises the possibility that, in some cases, diabetic embryopathy may be due to a mitochondrial cytopathy that affects both the mother's pancreas (and results in diabetes mellitus and the metabolic dysfunction associated with it) and the embryonic/fetal and placental tissues which make the embryo more vulnerable to this insult. PMID- 8723071 TI - VACTERL with the mitochondrial np 3243 point mutation. AB - The VACTERL association of vertebral, anal, cardiovascular, tracheo-esophageal, renal, and limb defects is one of the more common congenital disorders with limb deficiency arising during blastogenesis. The cause is probably heterogeneous; a molecular basis has not yet been defined. We report on a family in which a female infant with VACTERL was born in 1977 and died at age 1 month due to renal failure. Because her mother and sister later developed classical mitochondrial cytopathy associated with the A-G point mutation at nucleotide position (np) 3243 of mitochondrial (mt) DNA, we performed a molecular analysis of mt DNA in preserved kidney tissue from the VACTERL case. We discovered 100% mutant mt DNA in multicystic and 32% mutant mt DNA in normal kidney tissue. Mild deficiency of complex I respiratory chain enzyme activity was found in the mother's muscle biopsy. Other maternal relatives were healthy but had low levels of mutant mt DNA in blood. This is the first report to provide a precise molecular basis for a case of VACTERL. The differing tissue pathology depending on the percentage of mutant mt DNA suggests a causal connection between the mutation and symptoms. VACTERL, and this type of multicystic renal dysplasia, are new phenotypes for the np 3243 point mutation. The possibility of a mitochondrial disorder should be born in mind and also that VACTERL may occur as a first manifestation of a mutation that has been present for generations. This would have major implications for patient management and for genetic counselling regarding both the risk of recurrence and risk of other mitochondrial syndromes in affected families. PMID- 8723073 TI - Hand anomalies in fetal-hydantoin syndrome: from nail/phalangeal hypoplasia to unilateral acheiria. PMID- 8723074 TI - Exclusion of 22q11 deletion in Noonan syndrome with tetralogy of Fallot. PMID- 8723075 TI - Recurrent anencephaly as a primary manifestation of the acrocallosal syndrome. PMID- 8723076 TI - Revised diagnostic criteria for the Marfan syndrome. AB - In 1986, the diagnosis of the Marfan syndrome was codified on the basis of clinical criteria in the Berlin nosology [Beighton et al., 1988]. Over time, weaknesses have emerged in these criteria, a problem accentuated by the advent of molecular testing. In this paper, we propose a revision of diagnostic criteria for Marfan syndrome and related conditions. Most notable are: more stringent requirements for diagnosis of the Marfan syndrome in relatives of an unequivocally affected individual; skeletal involvement as a major criterion if at least 4 of 8 typical skeletal manifestations are present; potential contribution of molecular analysis to the diagnosis of Marfan syndrome; and delineation of initial criteria for diagnosis of other heritable conditions with partially overlapping phenotypes. PMID- 8723077 TI - A split hand-split foot (SHFM3) gene is located at 10q24-->25. AB - The split hand-split foot (SHSF) malformation affects the central rays of the upper and lower limbs. It presents either as an isolated defect or in association with other skeletal or non-skeletal abnormalities. An autosomal SHSF locus (SHFM1) was previously mapped to 7q22.1. We report the mapping of a second autosomal SHSF locus to 10q24-->25. A panel of families was tested with 17 marker loci mapped to the 10q24-->25 region. Maximum lod scores of 3.73, 4.33 and 4.33 at a recombination fraction of zero were obtained for the loci D10S198, PAX2 and D10S1239, respectively. An 19 cM critical region could be defined by haplotype analysis and several genes with a potential role in limb morphogenesis are located in this region. Heterogeneity testing indicates the existence of at least one additional autosomal SHSF locus. PMID- 8723078 TI - Evolution of the bone dysplasia family. PMID- 8723079 TI - Osteochondrodysplasias in South Africa. AB - Jurgen Spranger's visit to the University of Cape Town in 1974 provided impetus for the establishment of a bone dysplasia registry. By 1996 more than 2,500 affected persons had been documented and radiographs and DNA had been obtained in many instances. Of these disorders, about 1,500 fall into the category of "osteochondrodysplasias" as listed in the International Nomenclature [Spranger, 1992]. The numbers of affected persons with each of these disorders are presented in this article. Departmental or collaborative investigations on DNA banked in conjunction with the registry, has resulted in localization or characterization of several determinant genes. In this way, Spranger's early contributions to the Cape have led directly to the elucidation of several import genetic skeletal dysplasias. PMID- 8723080 TI - Stuve-Wiedemann syndrome: update and historical footnote. AB - Stuve-Wiedemann syndrome (SWS) is, at last, beginning to emerge from the shadows of campomelic syndrome as a nosologically and, presumably, causally-distinct entity, first delineated in 1971 on the basis of 2 affected sisters. The fact that these sisters had an affected double first cousin supports autosomal recessive inheritance of SWS. PMID- 8723081 TI - Stuve-Wiedemann dysplasia in a 3 1/2-year-old boy. AB - Stuve-Wiedemann osteochondrodysplasia is a rare disorder with distinct clinical and diagnostic radiographic findings. The condition is classified as a bent-bone dysplasia with early, lethal outcome. We report on a boy with Stuve-Wiedemann syndrome who is well and alive at the age of 3 1/2 years. PMID- 8723082 TI - Sponastrime dysplasia: five new cases and review of nine previously published cases. AB - Sponastrime dysplasia (SD) is a dwarfing autosomal recessive short-limb bone dysplasia. The diagnosis is established by a combination of clinical and radiological findings of which the radiological are the more specific. The current diagnostic criteria are ambiguous as demonstrated by the fact that, in our opinion, three of the five patients reported since the original article do not have this condition. Comparison of our five patients and the 9 published patients has led to development of more specific diagnostic criteria. Previously undescribed complications of this condition are subglottic stenosis and tracheo broncho-malacia, developmental coxa vara, and avascular necrosis of the capital femoral epiphyses. PMID- 8723083 TI - Diastrophic dysplasia: extreme variability within a sibship. AB - Three sibs were eventually diagnosed as having diastrophic dysplasia. The firstborn, a male, lacked many of the clinical manifestations and radiological abnormalities of the hands, such that the diagnosis of diastrophic dysplasia could not be made. Subsequently, his twin sisters were born with more severe skeletal involvement, but not many additional clinical findings, until one of his sisters (patient 2) developed a left ear cyst at age 2 months. Some cases of diastrophic dysplasia may be mild enough not to allow for a secure diagnosis, particularly in a neonate or infant occurring as the first affected person in the family. PMID- 8723084 TI - Kniest dysplasia: radiologic, histopathological, and scanning electronmicroscopic findings. AB - We describe severe neonatal Kniest dysplasia. Radiological findings in a severe case include short bowed tubular bones with exaggerated metaphyseal flare, moderate platyspondyly with vertical clefts of the vertebral bodies, and characteristically shaped iliac bones. Pathologic findings included a disorganized physeal growth plate, soft crumbly cartilage with a "Swiss-cheese" appearance, and diastase resistant intracytoplasmic inclusions in the resting chondrocytes. Transmission electronmicroscopy showed dilated cisternae of rough endoplasmic reticulum with finely granular material of accumulated protein. Scanning electronmicroscopy documented striking fragmentation and disintegration of collagen fibrils resulting in a web-like pattern and large open cyst-like spaces, and deficiency and disorganization of the collagen fibrils. PMID- 8723085 TI - Dyssegmental dysplasia with glaucoma. AB - We report on a "new" syndrome in 2 unrelated children with some manifestations of Kniest dysplasia and with spine abnormalities suggestive of dyssegmental dysplasia. Glaucoma with important ocular impairment was associated with severe dwarfism. No mutation of the COL2A1 gene was detected. The inheritance of this new type of skeletal dysplasia in unknown. PMID- 8723086 TI - Clinical and ultrastructural findings in three patients with geleophysic dysplasia. AB - Geleophysics dysplasia, a rare disorder with autosomal-recessive inheritance, is characterized by short stature with a "happy-looking" facial appearance. Nonskeletal findings, particularly in an advanced stage, include hepatosplenomegaly and valvular cardiopathy. Based on the clinical picture and the detection of lysosome-like inclusions in hepatocytes, the underlying cause of the condition is considered to be a storage defect in the metabolism of glycoproteins. The clinical course, with progressive worsening of the condition favors this hypothesis. We report on 3 further cases, in which light and electron microscopic studies of iliac crest biopsies and cultured skin fibroblasts provided additional evidence that geleophysic dysplasia represents a lysosomal storage disease. The additional discovery of storage vacuoles in chondrocytes and skin fibroblasts strongly suggests that the condition is a generalized storage defect. To date, it has not yet been possible to identify the presumed biochemical defect in the metabolic pathways of glycoproteins. PMID- 8723087 TI - Syndrome of multiple epiphyseal dysplasia (ribbing type) with rhizomelic shortness, cleft palate, and micrognathia in two unrelated patients. AB - We report on two unrelated patients with an apparently new syndrome. In each family they are the only affected members, their parents are not consanguineous, and paternal and maternal ages are not advanced. At birth each patient was noted to have a marked Robin phenotype (cleft of the secondary palate and micrognathia) plus rhizomelic shortness. Delay in the appearance of long bone epiphyses was noted and followed by small fragmented and later very flat epiphyses of all long bones. The fibulae are short and radial heads dislocated. Scoliosis and marked genu valgum developed in both. Both patients have normal intelligence, vision, and hearing. Both have mildly upward slanting palpebral fissures, broad nasal tip, and apparent hypertelorism. PMID- 8723088 TI - Metaphyseal peg in geroderma osteodysplasticum: a new genetic bone marker and a specific finding? AB - We describe two sibs with geroderma osteodysplasticum (GO) who, in addition to the known clinical and radiologic manifestations of the disorder, presented a metaphyseal peg indenting the epiphysis of the long bones, particularly at the knees. The peg was visible only at the age of 4 to 5 years but was invisible in infancy and following physeal closure. This may explain why this anomaly was not described in previous reports of 23 patients in 11 families with GO. The metaphyseal peg is an abnormality of bone development so far unknown to us. We speculate that it represents a primary, agedependent alteration of bone shape and hence a new genetic bone marker apparently specific to GO. PMID- 8723089 TI - Osteopathia striata with cranial sclerosis: variable expressivity in a four generation pedigree. AB - Osteopathia striata is a manifestation of several bone dysplasias. In association with cranial sclerosis it represents a separate entity, which is not limited to the bones but may affect other structures, leading to abnormal face, cleft palate, deafness, heart defects, and vertebral anomalies. Neurological findings range from normal development to marked retardation with hydrocephalus, cranial nerve deficiencies and deafness. Ten families, including our own, clearly demonstrate autosomal dominant inheritance with female preponderance and great inter- and intrafamilial variability. PMID- 8723090 TI - Jeune syndrome with tongue lobulation and preaxial polydactyly, and Jeune syndrome with situs inversus and asplenia: compound heterozygosity Jeune-Mohr and Jeune-Ivemark? AB - We report on a male infant with internal hydrocephalus, absence of corpus callosum, papillomas and lobulation of the tongue, notches of the alveolar ridges, short ribs, dysplastic pelvis, hypospadias, short limbs with bowed long tubular bones and postaxial polydactyly of hands, and preaxial polydactyly in one foot. Radiologically this case shares manifestations with Jeune syndrome; the tongue lobulation and the preaxial polydactyly are similar to findings in Mohr syndrome, or short-rib polydactyly syndrome (SRPS), type Majewski. In addition, a female newborn presented with manifestations of Jeune and Ivemark syndromes. One explanation for this overlap may be compound heterozygosity for these syndromes. PMID- 8723091 TI - An unknown spondylo-meta-epiphyseal dysplasia in sibs with extreme short stature. AB - In three sibs of Jordanian descent a unique type of severe spondylo-meta epiphyseal dysplasia results in extreme disproportionate dwarfism. They have a distinct facial appearance with hypotelorism, prognathia, and hypodontia. The limbs are short and the hands and feet stubby. Radiologically, the irregular end plates of the vertebral bodies, the very small and late appearing epiphyseal ossification centres, and the hypoplastic acetabular roofs are most impressive. Histopathologic studies of the growth plate demonstrate characteristic findings with fingerprint-like inclusion bodies in the hypertrophic chondrocytes. This seems to be a distinct, autosomal recessive skeletal dysplasia. PMID- 8723092 TI - Blomstrand lethal osteochondrodysplasia. AB - We present the clinical, roentgenographic, and histologic abnormalities in a stillborn infant with Blomstrand osteochondrodysplasia. Parental consanguinity and multiplex occurrence in the patients' sibship confirm the hypothesis of autosomal recessive inheritance of this monogenic lethal entity. The unknown genetic defect interferes severely with skeletal growth through lack of chondrocyte multiplication and apparent uncoupling of the processes of enchondral ossification and skeletal growth. PMID- 8723094 TI - Progress in medical genetics: map-based gene discovery and the molecular pathology of skeletal dysplasias. PMID- 8723093 TI - Prenatal diagnosis of osteochondrodysplasias in high risk pregnancy. AB - We collected data on 39 prenatally diagnosed osteochondrodysplasias. We detected 30 (76.9%) cases in the first and second trimesters, including 18 (46.2%) with two twins before the 24th week of gestation. Of 39 cases 11 (28.2%) had osteogenesis imperfecta (OI) type II. Verification of the prenatal diagnosis was attempted in 26 cases on the basis of the data obtained from ultrasonographs, radiographs, external examination, and autopsy protocols. The prenatal diagnosis was confirmed in 19 (73%) fetuses. In 13 cases verification was not possible because one or several investigations could not be performed. Counselling followed all identified cases with osteochondrodysplasia. We present the pedigree of two families indicating the possibility of early prenatal diagnosis of achondrogenesis type I and metatropic dysplasia. We propose indications for ultrasonographic anatomical screening with subsequent phenotype analysis in high risk pregnancy to provide for the prenatal detection of malformations and hereditary diseases. PMID- 8723095 TI - Molecular defects in the chondrodysplasias. AB - There has been a recent explosion of knowledge concerning the biochemical and molecular defects in the skeletal dysplasias. Through both the candidate gene approach and positional cloning, specific gene defects that produce the skeletal dysplasias have been identified and may be classified into several general categories: 1) qualitative or quantitative abnormalities in the structural proteins of cartilage; 2) inborn errors of cartilage metabolism; 3) defects in local regulators of cartilage growth; and 4) systemic defects influencing cartilage development. PMID- 8723096 TI - Phenotypic expressions of a Gly 154Arg mutation in type II collagen in two unrelated patients with spondyloepimetaphyseal dysplasia (SEMD). AB - Type II collagenopathies consist of chondrodysplasias ranging from lethal to mild in severity. A large number of mutations has been found in the COL2A1 gene. Glycine substitutions have been the most common types of mutation. Genotype phenotype correlations in type II collagenopathies have not been established, partly because of insufficient clinical and radiographic description of the patients. We found a glycine-to-arginine substitution at position 154 in type II collagen in two unrelated isolated propositi with spondyloepimetaphyseal dysplasia and provide a comparative clinical and radiographic analysis from birth to young adulthood for this condition. The clinical phenotype was disproportionate short stature with varus/valgus deformities of the lower limbs requiring corrective osteotomies, and lumbar lordosis. The skeletal radiographs showed an evolution from short tubular bones, delayed epiphyseal development, and mild vertebral involvement to severe metaphyseal dysplasia with dappling irregularities, and hip "dysplasia." The metaphyseal abnormalities disappeared by adulthood. PMID- 8723097 TI - A specific collagen type II gene (COL2A1) mutation presenting as spondyloperipheral dysplasia. AB - We report on a patient with a skeletal dysplasia characterized by short stature, spondylo-epiphyseal involvement, and brachydactyly E-like changes. This condition has been described as spondyloperipheral dysplasia and the few published cases suggest autosomal dominant inheritance with considerable clinical variability. We found our sporadic case to be due to a collagen type II defect resulting from a specific COL2A1 mutation. This mutation is the first to be located at the C terminal outside the helical domain of COL2A1. A frameshift as consequence of a 5 bp duplication in exon 51 leads to a stop codon. The resulting truncated C propeptide region seems to affect helix formation and produces changes of chondrocyte morphology, collagen type II fibril structure and cartilage matrix composition. Our case with its distinct phenotype adds another chondrodysplasia to the clinical spectrum of type II collagenopathies. PMID- 8723098 TI - An alpha 1(II) Gly913 to Cys substitution prevents the matrix incorporation of type II collagen which is replaced with type I and III collagens in cartilage from a patient with hypochondrogenesis. AB - A heterozygous mutation in the COL2A1 gene was identified in a patient with hypochondrogenesis. The mutation was a single nucleotide transition of G3285T that resulted in an amino acid substitution of Cys for Gly913 in the alpha 1(II) chain of type II collagen. This amino acid change disrupted the obligatory Gly-X Y triplet motif required for the normal formation of a stable collagen triple helix and prevented the deposition of type II collagen into the proposita's cartilage, which contained predominantly type I and III collagens and minor amounts of type XI collagen. Biosynthetic analysis of collagens produced and secreted by the patient's chondrocytes cultured in alginate beads was consistent with the in vivo matrix composition, demonstrating that the main products were type I and III collagens, along with type XI collagen. The synthesis of the cartilage-specific type XI collagen at similar levels to controls indicated that the isolated cartilage cells had re-differentiated to the chondrocyte phenotype. The chondrocytes also produced small amounts of type II collagen, but this was post-translationally overmodified and not secreted. These data further delineate the biochemical and phenotypic consequences of mutations in the COL2A1 gene and suggest that cartilage formation and bone development can take place in the absence of type II collagen. PMID- 8723099 TI - Type II collagenopathies: are there additional family members? AB - The type II collagenopathies represent a group of chondrodysplasias sharing clinical and radiological manifestations which are expressed as a continuous spectrum of phenotypes, ranging from perinatally lethal to very mild conditions. Their common molecular bases are mutations in the type II collagen gene (COL2A1). We describe one case of lethal platyspondylic dysplasia, Torrance type, and a variant of lethal Kniest dysplasia, neither of which has been reported as a type II collagenopathy. Biochemical studies of cartilage collagens and morphological analysis of cartilage sections suggest that abnormalities of type II collagen structure and biosynthesis are the main pathogenetic factors in both cases. Thus, the phenotypic spectrum of type II collagenopathies might be greater than hitherto suspected. PMID- 8723100 TI - A chondrodysplasia family produced by mutations in the diastrophic dysplasia sulfate transporter gene: genotype/phenotype correlations. AB - Achondrogenesis type 1B (ACG-1B), atelosteogenesis type 2 (AO-2), and diastrophic dysplasia (DTD) are recessively inherited chondrodysplasias of decreasing severity caused by mutations in the diastrophic dysplasia sulfate transporter (DTDST) gene on chromosome 5. In these conditions, sulfate transport across the cell membrane is impaired which results in insufficient sulfation of cartilage proteoglycans and thus in an abnormally low sulfate content of cartilage. The severity of the phenotype correlates well with the predicted effect of the underlying DTDST mutations: homozygosity or compound heterozygosity for stop codons or transmembrane domain substitutions mostly result in achondrogenesis type 1B, while other structural or regulatory mutations usually result in one of the less severe phenotypes. The chondrodysplasias arising at the DTDST locus constitute a bone dysplasia family with recessive inheritance. PMID- 8723101 TI - Common mutations in the fibroblast growth factor receptor 3 (FGFR 3) gene account for achondroplasia, hypochondroplasia, and thanatophoric dwarfism. AB - The mapping of the achondroplasia locus to the short arm of chromosome 4 and the subsequent identification of a recurrent missense mutation (G380R) in the fibroblast growth factor receptor 3 (FGFR-3) gene has been followed by the detection of common FGFR-3 mutations in two clinically related disorders: thanatophoric dwarfism (types I and II) and hypochondroplasia. The relative clinical homogeneity of achondroplasia was substantiated by demonstration of its genetic homogeneity as more than 98% of all patients hitherto reported exhibit mutations in the transmembrane receptor domain. Although most hypochondroplasia cases were accounted for by a recurrent missense substitution (N540K) in the first tyrosine kinase (TK 1) domain of the receptor, a significant proportion (40%) of our patients did not harbor the N540K mutation and three hypochondroplasia families were not linked to the FGFR-3 locus, thus supporting clinical heterogeneity of this condition. In thanatophoric dwarfism (TD), a recurrent FGFR-3 mutation located in the second tyrosine kinase (TK 2) domain of the receptor was originally detected in 100% of TD II cases, our series seven distinct mutations in three different protein domains were identified in 25 of 26 TD I patients, suggesting that TD, like achondroplasia, is a genetically homogenous skeletal disorder. PMID- 8723102 TI - Radiological and histological variants of thanatophoric dysplasia are associated with common mutations in FGFR-3. AB - We describe two fetuses of the 21st week of gestation that share some macroscopic, radiologic, and histologic findings of thanatophoric dysplasia (TD), but also show distinct differences from the usual subtypes of TD. These differences mainly comprise the lack of facial abnormality, only mild reduction of chondrocyte proliferation and hypertrophy, and the lack of fibrous tissue interposition between cartilage and periosteal bone. Thus, these two cases may represent a distinct variant of thanatophoric dysplasia. The molecular analysis of the FGF-R-3 gene demonstrated in both cases mutations which were not significantly different from those of other cases of TD. Thus, the phenotypic modulation within the subtypes of TD may be influenced by additional and yet unknown factors. PMID- 8723103 TI - Deficient expression of the small proteoglycan decorin in a case of severe/lethal osteogenesis imperfecta. AB - In osteogenesis imperfecta (OI) the effects of mutations in type I collagen genes generally reflect their nature and localization. Unrelated individuals sharing identical mutations present, in general, similar clinical phenotypes. However, in some such cases the clinical phenotype differs. This variable clinical expression could be the result of abnormalities in other connective tissue proteins. Since decorin is a component of connective tissue, binds to type I collagen fibrils and plays a role in matrix assembly, we studied decorin production in skin fibroblasts from OI patients. Cultured fibroblasts from one patient with extremely severe osteogenesis imperfecta (classified as type II/III) who has an alpha 1(I)gly415ser mutation were found to secrete barely detectable amounts of decorin into culture medium. Western blotting using antibodies raised against decorin confirmed the reduction of the decorin core protein and Northern blot analysis showed decorin mRNA levels below the limit of detection. Cells from a patient, with a less severe phenotype, bearing a mutation in the same position of the triple helix (alpha 1(I)gly415) expressed decorin normally. The different clinical phenotypes could be due to the differing genetic backgrounds of the patients so it is tempting to conclude that in our most severely affected patient the absence of decorin aggravates the clinical phenotype. PMID- 8723105 TI - Asymmetrical coronal synostosis, cutaneous syndactyly of the fingers and toes, and jejunal atresia in a male infant. AB - We report on a male infant with an unusual type of acrocephalosyndactyly presenting with unilateral coronal craniosynostosis, cutaneous syndactyly of toes 2 and 3, loss of distal triradii, and transverse alignment of dermal ridges of the palm suggesting syndactyly, atresia of the proximal jejunum, and anal stenosis. PMID- 8723104 TI - Tracing the pathway between mutation and phenotype in osteogenesis imperfecta: isolation of mineralization-specific genes. AB - The brittleness of bone in people with lethal (type II) osteogenesis imperfecta, a heritable disorder caused by mutations in the type I collagen genes, arises from the deposition of abnormal collagen in the bone matrix. The inability of the abnormal collagen to participate in mineralization may be caused by its failure to interact with other bone proteins. Here, we have designed a strategy to isolate the genes important for mineralization of collagen during bone formation. Cells isolated from 16-day embryonic chick calvaria and seeded post-confluence in culture deposited a mineralized matrix over a period of 2 weeks. Chick skin fibroblasts seeded and cultured under the same conditions did not mineralize. Using RT-PCR, we prepared short cDNAs (approximately 300 bp) corresponding to the 3' ends of mRNA from fibroblasts and separately from the mineralizing calvarial cells. Subtractive cDNA hybridization generated a pool of cDNAs that were specific to mineralizing calvarial cells but not to fibroblasts. Screening of 100,000 plaques of a chick bone ZAP Express cDNA library with this pool of mineralizing-specific cDNAs identified ten clones which comprised full-length cDNAs for the bone proteins osteopontin (eight of the ten positives), bone sialoprotein II (one of the ten positives), and cystatin (one of the ten positives). cDNAs for type I collagen, fibronectin, alkaline phosphatase, house keeping genes, and other genes expressed in fibroblasts were not identified in this preliminary screen. The pool of short cDNAs is likely to comprise cDNAs for further bone-specific genes and will be used to screen the entire bone cDNA library of 4.2 million clones. PMID- 8723106 TI - Craniosynostosis suggestive of Saethre-Chotzen syndrome: clinical description of a large kindred and exclusion of candidate regions on 7p. AB - We describe the clinical manifestations of an autosomal dominant form of craniosynostosis in a large family with eight affected relatives. Unilateral or bilateral coronal synostosis, low frontal hair line, strabismus, ptosis, and partial cutaneous syndactyly of fingers and toes are findings suggestive of the diagnosis of Saethre-Chotzen syndrome. The disease locus was excluded from the two adjacent Saethre-Chotzen candidate regions on 7p by linkage analysis with markers D7S664 and D7S507. This indicates heterogeneity of Saethre-Chotzen syndrome with a locus outside the candidate regions on 7p. PMID- 8723107 TI - Ectrodactyly and absence (hypoplasia) of the tibia: are there dominant and recessive types? AB - We present a kindred of brother, sister, and cousin with ectrodactyly and hypoplasia of the tibia. The parents of the cousin are consanguineous; the parents of the sibs originate from the same small Algerian village. We also report on a boy with tibial defect and split hands and feet with consanguineous parents. These observations are further hints for an autosomal recessive type of ectrodactyly with aplasia (hypoplasia) of the tibia, as was favoured by some authors. However, review of the present and reported cases does not demonstrate any clinical differences between the seemingly recessive and the dominant types. Statistical analysis of 17 families with affected sibs and normal parents showed a 1:3.1 ratio of affected:unaffected by the proband method. Despite consanguinity among nine sets of parents, this ratio, and approximately 30 additionally reported families generally are in favour of autosomal dominance with reduced penetrance. PMID- 8723108 TI - Rectal atresia as rare manifestation in EEC syndrome. AB - A newborn boy presented with bilateral split hand/foot malformation, sparse hair, dry and scaly skin, and nasolacrimal duct obstruction. Despite absence of cleft lip or palate, the findings fit the EEC syndrome. Additionally, the boy had rectal atresia. At least six further patients with EEC syndrome and anal atresia (two published, four unpublished) demonstrate, that anorectal malformation is a further, but rare anomaly in EEC syndrome. PMID- 8723109 TI - Bilateral radial deficiency with lower limb involvement. AB - We describe a 10-month-old boy with an unclassified form of radial aplasia with absent thumbs, tibia hypo/-aplasia, and partial absence of toes. Only a few cases with similar limb deficiencies have been published. We try to classify the malformations on the basis of embryological considerations and discuss possible differential diagnosis. PMID- 8723110 TI - Methylamine accumulation in cultured cells as a measure of the aqueous storage compartment in the laboratory diagnosis of genetic lysosomal diseases. AB - Intracellular accumulation of the lysosomotropic compound [14C]methylamine was used to estimate the size of the lysosomal compartment in fibroblasts cultured from patients with a variety of lysosomal storage diseases. In previous work from our laboratory, it was shown that methylamine accumulation was significantly increased in diseases with infantile or juvenile onset and storage of predominantly water-soluble material such as in the mucopolysaccharidoses, mucolipidoses, and oligosaccharidoses. In the present study, methylamine incorporation was abnormally increased in cells from patients with glycogenosis type II and with Niemann-Pick type C disease, whereas it was normal in other sphingolipidoses and in the late-infantile and juvenile forms of neuronal ceroid lipofuscinoses. The methylamine test was also checked regarding its potential use for prenatal diagnostic testing. In model systems with cultured amniotic or chorionic villus cells, lysosomal storage was experimentally induced by the cathepsin inhibitor leupeptin and was readily detected when compared to untreated controls. Cultured amniotic cells from a fetus with mucopolysaccharidosis II were found to incorporate significantly higher amounts of [14C]methylamine than the normal controls. The results indicate that the methylamine accumulation method is an additional tool in the diagnosis and prenatal diagnosis of lysosomal diseases with abnormal storage of water-soluble material. PMID- 8723111 TI - The spectrum of free neuraminic acid storage disease in childhood: clinical, morphological and biochemical observations in three non-Finnish patients. AB - N-acetylneuraminic acid (sialic acid) storage disease is a rare autosomal recessive lysosomal disorder. Clinically two major forms exist, an infantile type with severe progression leading to early death, and a milder form (Salla disease) with a protracted course. Intermediate forms may also exist. Diagnosis rests on the determination of an excessive excretion of sialic acid in urine and concomitant storage in fibroblasts, the severe forms exhibiting the highest excretion and storage. We present clinical, morphological, and biochemical data on three non-Finnish patients with sialic acid storage disease. Patient 1 was a preterm infant with neonatal ascites, coarse face, hepatosplenomegaly, pale skin, and wispy hair. Vacuolated lymphocytes were abundant in a peripheral blood smear and he excreted large amounts of free sialic acid. High levels of free sialic acid were also found in cultured skin fibroblasts. He died at age 6 months from progressive respiratory insufficiency. Patient 2 was an 11-month-old Egyptian girl with coarse face, frequent upper respiratory tract infections, hepatosplenomegaly, and severe psycho-motor retardation. Sialic acid excretion was elevated, likewise the storage in fibroblasts. Histological investigations documented vacuolar storage in a skin biopsy and in iliac crest tissue. Patient 3 was a 16-year-old girl with slightly coarse face, severe generalized muscular hypotonia, ataxia, and kyphoscoliosis originally diagnosed as having post-partum asphyxia. She suffered progressive motor function loss and had dysarthria. Urinary sialic acid was elevated and a skin biopsy demonstrated vacuolization. The clinical variability of sialic acid storage disease is exemplified by these three cases. Simple urinary screening for free sialic acid facilitates the diagnosis. The degree of urinary excretion may indeed correlate with clinical presentation and progression. PMID- 8723112 TI - Clinical and neuroradiological findings in classic infantile and late-onset globoid-cell leukodystrophy (Krabbe disease). AB - In the present study the clinical course and imaging of early and late-onset forms of Krabbe disease are analyzed. We report on 11 patients with a biochemical diagnosis of galactosyl ceramide beta-galactoside deficiency. Two presented as the classic infantile form and died within the second year of life. In 9 children the first clinical signs, such as gait difficulties and visual failure, started after age 2 years. All these patients developed slow regression of motor and mental capacities, and most of them died within their first decade. In patients of both groups computed tomography (CT) and magnetic resonance imaging (MRI) were performed. In the late-onset form, hypodensities of the central white matter and pyramidal tracts were the leading radiological signs, whereas in the early-onset form, hyperdensities and cerebellar white matter lesions were also detected. From our results it becomes clear that variability of Krabbe disease refers not only to clinical manifestation but also to CT and MRI findings. Better knowledge of phenotypic and radiological diversity will help to understand the pathogenesis of the disease. PMID- 8723113 TI - The clinical and molecular spectrum of androgen insensitivity syndromes. AB - Androgen insensitivity syndromes (AIS) are due to end-organ resistance to androgenic steroids in males leading to defective virilization of the external genitalia. The phenotype encompasses a wide array of genital ambiguity and may range from completely female to undervirilized but unequivocally male with infertility. This disorder is caused by mutations of the androgen receptor and is an X-linked recessive trait. We have studied 47 patients with AIS and have characterized the underlying molecular abnormality in the androgen receptor gene. Twenty patients had complete AIS and twenty-seven had partial AIS. Of the latter, 11 were of predominantly female phenotypic appearance and gender was assigned accordingly, while 16 were raised as males. Within the group of complete AIS, two patients had gross deletions within the gene, one had a small deletion, and one had an insertion. In the other patients with complete AIS, as well as all individuals with partial AIS, single nucleotide substitutions within the coding region were detected, each leading to an amino acid alteration. Seven codons were involved in more than one mutation in different cases. In addition, in one patient with spinal and bulbar muscular atrophy, an elongation of a glutamine repeat was characterized. We conclude that mutations in the androgen receptor gene may be present throughout the whole coding region. However, our study provides evidence that several mutational hot spots exist. PMID- 8723114 TI - Phenotypic classification of male pseudohermaphroditism due to steroid 5 alpha reductase 2 deficiency. AB - Conversion of testosterone (T) to dihydrotestosterone (DHT) in genital tissue is catalysed by the enzyme 5 alpha-reductase 2, which is encoded by the SRD5A2 gene. The potent androgen DHT is required for full masculinization of the external genitalia. Mutations of the SRD5A2 gene inhibit enzyme activity, diminish DHT formation, and hence cause masculinization defects of varying degree. The classical syndrome, formerly described as pseudovaginal perineoscrotal hypospadias, is characterized by a predominantly female phenotype at birth and significant virilization without gynecomastia at puberty. We investigated nine patients with steroid 5 alpha-reductase 2 deficiency (SRD). Phenotypes, which were classified according to the severity of the masculinization defect, varied between completely female (SRD type 5), predominantly female (SRD type 4), ambiguous (SRD type 3), predominantly male with micropenis and hypospadias (SRD type 2), and completely male without overt signs of undermasculinization (SRD type 1). T/DHT-ratios were highly increased ( > 50) in the classical syndrome (SRD type 5), but variable in the less severe affected patients (SRD types 1-4) (14-35). Mutations in the SRD5A2 gene had been characterized using PCR-SSCP analysis and direct DNA sequencing. A small deletion was encountered in two patients, while all other patients had single base mutations which result in amino acid substitutions. We conclude that phenotypes may vary widely in patients with SRD5A2 gene mutations spanning the whole range from completely female to normal male without distinctive clinical signs of the disease. Hence, steroid 5 alpha-reductase deficiency should be considered not only in sex reversed patients with female or ambiguous phenotypes, but also in those with mild symptoms of undermasculinization as encountered in patients with hypospadias and/or micropenis. A classification based on the severity of the masculinization defect may be used for correlation of phenotypes with enzyme activities and genotypes, and for comparisons of phenotypes between different patients as the basis for clinical decisions to be made in patients with pseudohermaphroditism due to steroid 5 alpha-reductase 2 deficiency. PMID- 8723115 TI - Differential display RT PCR of total RNA from human foreskin fibroblasts for investigation of androgen-dependent gene expression. AB - Male sexual differentiation is a process that involves androgen action via the androgen receptor. Defects in the androgen receptor, many resulting from point mutations in the androgen receptor gene, lead to varying degrees of impaired masculinization in chromosomally male individuals. To date no specific androgen regulated morphogens involved in this process have been identified and no marker genes are known that would help to predict further virilization in infants with partial androgen insensitivity. In the present study we first show data on androgen regulated gene expression investigated by differential display reverse transcription PCR (dd RT PCR) on total RNA from human neonatal genital skin fibroblasts cultured in the presence or absence of 100 nM testosterone. Using three different primer combinations, 54 cDNAs appeared to be regulated by androgens. Most of these sequences show the characteristics of expressed mRNAs but showed no homology to sequences in the database. However 15 clones with significant homology to previously cloned sequences were identified. Seven cDNAs appear to be induced by androgen withdrawal. Of these, five are similar to ETS (expression tagged sequences) from unknown genes; the other two show significant homology to the cDNAs of ubiquitin and human guanylate binding protein 2 (GBP-2). In addition, we have identified 8 cDNA clones which show homologies to other sequences in the database and appear to be upregulated in the presence of testosterone. Four of these clones again are similar to ETS from unknown genes. Three differential expressed sequences that appear to be upregulated in the presence of testosterone show significant homology to the cDNAs of L-plastin and one to the cDNA of testican. This latter gene codes for a proteoglycan involved in cell social behavior and therefore of special interest in this context. The results of this study are of interest in further investigation of normal and disturbed androgen-dependent gene expression. PMID- 8723117 TI - Clinical and molecular cytogenetic observations in three cases of "trisomy 12p syndrome". AB - Two unpublished cases with partial tandem duplication of 12p and one previously published case were studied by fluorescence in situ hybridization using 11 cosmid DNA probes from 12p. We propose that the smallest duplications of 12(p13.2pter) and 12(p13.1p13.33) produce the "trisomy 12p syndrome" which is characterized by heavy birth weight, macrocephaly, muscular hypotonia, short neck, flat face, high forehead, prominent cheeks, large philtrum, short nose with anteverted nostrils, and broad everted lower lip. From a review of the published cases we conclude that gross malformations are lacking in "pure" trisomy 12p, and mental retardation is severe in complete and moderate in partial trisomy 12p. Polydactyly and accessory nipples were found only with almost complete trisomy 12p. Abnormalities of hair growth may be related to a gene at 12p. The sub-band 12p11.21 may be critical for acrocallosal syndrome. Macrocephaly may be due to a metabolic disorder. PMID- 8723116 TI - Short stature in a mother and daughter with terminal deletion of Xp22.3. AB - Short stature in females is often caused by hemizygosity for the terminal portion of Xp due to monosomy X or a deletion. We report on a mother and daughter with short stature as sole phenotypic abnormality and deletion of bands Xp22.32-p22.33 demonstrated by classic and molecular cytogenetic analysis. In both individuals, the deleted X chromosome was late replicating. Molecular analysis suggested that the deletion is terminal and the breakpoint was localized between the STS and DXS7470 loci in Xp22.32. Chromosome analysis is often done on females with short stature to exclude Ullrich-Turner syndrome. Small deletions, terminal or interstitial, are easily missed by conventional cytogenetic investigation; thus molecular analyses are useful to detect those cases. PMID- 8723118 TI - Cytogenetic and molecular analysis in trisomy 12p. AB - We studied a male patient with de novo pure trisomy 12p syndrome by molecular analysis and fluorescence in situ hybridization (FISH) with markers from chromosome 12. G-banding studies demonstrated a 46,XY, 22p+ karyotype and the banding pattern and clinical findings suggested that the extra chromosomal material was derived from 12p. Trisomy 12p was confirmed by dosage analysis with chromosome 12p markers and FISH analysis with a whole chromosome 12 paint. The de novo re-arranged chromosome was of paternal origin. A comparison of the clinical and cytogenetic findings in this patient was made with previously described cases of trisomy 12p. We propose a classification system for 12p trisomy in order to better characterize the correlative relationships between specific cytogenetic constitution and phenotype. PMID- 8723119 TI - Computer assisted diagnosis of malformation syndromes: an evaluation of three databases (LDDB, POSSUM, and SYNDROC). AB - Computer programs which can be used as an aid to diagnose multiple congenital anomaly syndromes have been used for many years, but up to now they have been evaluated very rarely. The diagnostic abilities of three of these systems [LDDB (London Dysmorphology Database), POSSUM (Pictures of Standard Syndromes and Undiagnosed Malformations), and SYNDROC] were analyzed. All three programs are based on an algorithm which defines a diagnosis by a set of phenotypic components all having the same weight (descriptive algorithm). A second algorithm is applied by SYNDROC to rank competing diagnoses in order of probability. This pseudo Bayesian algorithm provides a coefficient of certitude (CC). For a test the clinical findings of 102 patients who had received a firm diagnosis were used. Two search strategies were tried: "novice's strategy" with all findings taken for a search and "expert's strategy" with a selected set of anomalies. Only those diagnoses that were suggested with the 1st rank, defined as the highest degree of agreement, or the highest CC were studied. The greatest resemblance between suggestions of the databases and the clinical diagnosis was obtained with the expert strategy. The highest number of matches were produced by SYNDROC (80 with expert strategy) and the lowest by POSSUM (54 with novice strategy). The overall agreement between the databases is about 40% for the 1st rank. This number reflects that different authors use different pivotal signs for the description of a syndrome. With the pseudo-Bayesian algorithm 59 cases obtained the highest CC value. Great difficulties exist with the subjective estimates for the calculation of these values; the absolute CC values seem to be meaningless. A small number of unusual cases with special combinations of anomalies provide serious problems for correct diagnosis. PMID- 8723120 TI - Serial examination of 20,248 newborn fetuses and infants: correlations between drug exposure and major malformations. AB - Maternal medication during the first trimester of pregnancy has been discussed as a risk factor for development of birth defects. The correlation between maternal drug use and major malformations was investigated in a population-based case control study in Mainz. Over a period of 5 years (1990-1994), 20,248 livebirths, stillbirths, and abortions underwent physical and sonographic examination, and anamnestic data were collected. A total of 1,472 births with congenital anomalies (cases) and 9,682 births without major and minor malformations (controls) were analyzed. We distinguished between 30 different drug categories, which were divided into medication taken continuously (before and during pregnancy; CM) and acute medication (drugs given within the first 3 months of gravidity; AM). Statistically highly-significant results [CM: Odds Ratios (OR) 1.2, Confidence Intervals (CI) 1.1-1.4, P = 0.008; AM: OR 1.2, CI 1.1-1.3, P = 0.008] were established for maternal drug use in correlation to birth defects. For the majority of combinations between drugs and specific malformations no teratogenic risks were found. However, statistically significant associations were recorded for antiallergics and heart anomalies (CM, AM) as well as musculoskeletal anomalies (AM); for bronchodilators and heart anomalies (CM, AM); for antiepileptics and anomalies of the internal urogenital system (CM), as well as cleft palate/cleft lips (AM); for thyroid hormones and anomalies of the nervous system (CM, AM), as well as anomalies of the external urogenital system (CM, AM); for insulin and anomalies of the musculoskeletal system (CM); for digitalis and anomalies of the musculoskeletal system (AM). PMID- 8723121 TI - Dubowitz syndrome: review of 141 cases including 36 previously unreported patients. AB - We review clinical information on 141 individuals with Dubowitz syndrome, 105 reported since 1965, and 36 previously unreported. We define the Dubowitz syndrome phenotype on the basis of clinical descriptions. The facial appearance is characteristic and present in most patients with Dubowitz syndrome. The phenotypic spectrum is quite variable and ranges from normal growth and head circumference with mild psychomotor retardation and lack of eczema to a condition of severe growth retardation, mental retardation, microcephaly, and eczema. Overall, the condition may involve the cutaneous, ocular, dental, digestive, musculoskeletal, urogenital, cardiovascular, neurological, hematological, and immune systems. Characteristic behavior patterns which have not been cited previously are present in our cases; most patients are hyperactive, shy, hate crowds, and like music, rhythm, and vibrations from music speakers, tape recorders, or transmitted through floors. Dubowitz syndrome is an autosomal recessive disorder with possibly increased frequency of parental consanguinity. Heterogeneity cannot be excluded at this time. PMID- 8723122 TI - Oculo-facio-cardio-dental (OFCD) syndrome. AB - Oculo-facio-cardio-dental syndrome consists of (1) eye anomalies: congenital cataract and microphthalmia, or secondary glaucoma, (2) facial abnormalities: (long narrow face, high nasal bridge, pointed nose with cartilages separated at the tip, cleft palate, or submucous cleft palate, (3) cardiac anomalies: atrial septal defect (ASD), ventricular septal defect (VSD), or floppy mitral valve; and (4) dental abnormalities: canine radiculomegaly, delayed dentition, oligodontia, persistent primary teeth, or variable root length. Other less common findings are: sensorineural hearing loss, septate vagina, and syndactyly of toes 2-3. Inheritance may be an X-linked dominant trait, lethal in the male. PMID- 8723123 TI - Arthrogryposis associated with unsuccessful attempts at termination of pregnancy. AB - We report three cases of failed termination of pregnancy in which the children were subsequently born with arthrogryposis (AMC) (multiple congenital contractures). Arthrogryposis is a sign with many causes. We suggest that the multiple congenital contractures seen in these children are due to vascular compromise during the attempted termination with secondary loss of functional neurons leading to fetal akinesia and subsequent contractures. Two of the children have additional evidence of intrauterine vascular compromise. Limitation of movement secondary to the rupture of the fetal membranes and continuous leakage of amniotic fluid after the attempted termination may have compounded the contractures in two of the children. PMID- 8723124 TI - Incidence and spectrum of renal abnormalities in Williams-Beuren syndrome. AB - Kidneys and urinary tract were examined systematically by ultrasonography in 130 patients with Williams-Beuren syndrome (59 females, median age 5.5 years; 71 males, median age 6.4 years). In addition, serum creatinine was determined and an analysis was performed. Creatinine clearance was available in 79 patients. Renal angiographic examinations were done in 18 patients, 8 of whom had renal artery narrowing (44%). The incidence of renal anomalies in Williams-Beuren syndrome was 17.7% vs. around 1.5% in the normal population (P < 0.0003). The spectrum of these anomalies ranged from minor anomalies such as bladder diverticula to more severe malformations such as renal aplasia or hypoplasia (in 5 of 130 patients). In nine patients a duplicated kidney was found. A decreased creatinine clearance (two patients), recurrent symptomatic urinary tract infections (four patients), and hypertension were uncommon. Nephrocalcinosis was not found in our patients. Our data demonstrate that the risk of a structural abnormality of the kidneys and the urinary tract is increased 12- to 36-fold in Williams-Beuren syndrome compared to the normal population. Ultrasound screening of the renal system should be part of the first evaluation of WBS patients. PMID- 8723125 TI - Informative morphogenetic and phenogenetic variants in children with cleft lip/cleft palate. AB - In 230 patients with nonsyndromic cleft lip/cleft palate (138 boys and 92 girls) and in 226 age related healthy children (137 boys and 89 girls) informative morphogenetic and phenogenetic variants (IMV and PHV, respectively) were investigated. There was no difference between the number of IMVs between both groups (chi 2 = 5.89; d.f. = 3; alpha > 0.70). This finding is in line with the hypothesis that facial cleft disorders occur during blastogenesis, whereas IMVs and PHVs are typical patterns of the embryo- and fetogenesis. The anthropometric findings are contradictory. In a few non-craniofacial phenogenetic variants significant differences were found between the patients and the healthy children. Intrinsic factors or secondary sequelae of the primary defect might additionally act in the morphological fine tuning of children with single cleft lip/cleft palate. PMID- 8723126 TI - Familial arhinia, choanal atresia, and microphthalmia. AB - We describe two females (aunt and niece) with variable manifestations of arhinia, choanal atresia, microphthalmia, and hypertelorism. In the literature there is only one report on this syndrome in sibs. We hypothesize autosomal dominant inheritance with reduced penetrance. PMID- 8723127 TI - Diffuse polymicrogyria associated with an unusual pattern of multiple congenital anomalies including turribrachycephaly and hypogenitalism. AB - A newborn male infant born to consanguineous parents was found to have diffuse polymicrogyria associated with an unusual pattern of congenital anomalies including microbrachycephaly, turricephaly, blepharophimosis, microstomia with maxillary retrusion and mandibular prognathism, micropenis with cryptorchidism, camptodactyly and adducted thumbs, and a progeroid appearance. The combination of manifestations in our patient represents a unique form of polymicrogyria with congenital anomalies, probably representing a new syndrome. PMID- 8723128 TI - Detection of a novel mutation in the SRC homology domain 2 (SH2) of Bruton's tyrosine kinase and direct female carrier evaluation in a family with X-linked agammaglobulinemia. AB - X-linked agammaglobulinemia (XLA) is an inherited immunodeficiency disease with a block in differentiation from pre-B to B cells resulting in a selective defect in the humoral immune response. Affected males have very low concentrations of serum immunoglobulins leading predominantly to recurrent bacterial infections beginning at age 6 to 18 months. The gene responsible for XLA was identified recently to encode a cytoplasmatic tyrosine kinase (Bruton's tyrosine kinase, BTK). We have analyzed the BTK gene in a large family in which two brothers presented with the severe phenotype of XLA. Genomic DNA of affected boys and from healthy relatives was amplified by PCR with primers specific for the putative promoter region and for all 19 exons, including flanking intron boundaries, and subsequently screened for mutations using single strand conformation polymorphism (SSCP) analysis. Altered single strand band patterns were found using primers specific for exon 10, 15, and 18. Direct cycle-sequencing of these BTK segments detected two known polymorphisms in intron 14 and in exon 18. Sequencing of exon 10 from two boys with XLA demonstrated a novel point mutation in the SH2 domain of BTK. Direct identification of healthy female carriers in three generations was performed by amplification mutagenesis using PCR with a modified first primer. This method can easily be applied also to prenatal diagnosis. PMID- 8723129 TI - Glomerulopathy associated with predominant fibronectin deposits: exclusion of the genes for fibronectin, villin and desmin as causative genes. AB - Glomerulopathy with predominant fibronectin deposits (GFD) is a newly recognized autosomal dominant renal disease that leads to albuminuria, microscopic hematuria, hypertension, renal tubular acidosis type IV, and end-stage renal disease in the second to fourth decade of life. Light microscopy documents extensive deposits in the subendothelial space, which on electron microscopy consist of non-oriented 12 x 125 nm fibers. Deposits are strongly immunoreactive for antibodies to fibronectin. We examined the hypothesis that a genetic defect in the gene for fibronectin is responsible for the disease. In a 197 member pedigree, 13 relatives developed end-stage renal failure from the disease. In 99 individuals haplotype analysis was performed using 6 microsatellite markers spanning a > 56 cM interval in chromosome region 2q34, where fibronectin, villin, and desmin map in close proximity. Haplotype analysis resulted in exclusion of the whole range of 78 cM covered by the markers examined. This result excludes fibronectin, villin, and desmin from being the causative genes for GFD in this large kindred. PMID- 8723130 TI - Enhanced synaptophysin immunoreactivity in rat hippocampal culture by 5-HT 1A agonist, S100b, and corticosteroid receptor agonists. AB - Serotonin (5-HT) has been shown to modulate brain maturation during development and adult plasticity. This effect in the whole animal may be due to activation of 5-HT1A receptors and a corresponding increases in S100b and corticosterone. Synaptophysin, an integral protein of the synaptic vesicle membrane that correlates with synaptic density and neurotransmitter release, is reduced by depletion of 5-HT in the cortex and hippocampus of the adult rat. Injections of a 5-HT1A agonist or dexamethasone can reverse the loss of synaptophysin immunoreactivity (IR). In this study we used morphometric analysis of synaptophysin-IR to study the effects of the 5-HT1A agonist, ipsapirone, and the neuronal extension factor, S100b on hippocampal neurons grown in a serum and steroid free media. Both compounds increased the synaptophysin-IR at doses previously established to be highly specific. Ipsapirone (10(-9)M) was more effective on neuronal cell bodies staining and S100b (10 ng/ml) was more effective in increasing the number of synaptophysin-IR varicosities on neuronal processes. In addition both types of corticosteroid receptor agonists, at previously established specific doses, Ru28362 (10(-8) M) and aldosterone (10(-9) M) produced smaller increases compared to control groups in both the cell body staining and the number of varicosities. The effect of these differentiating factors on the expression of synaptophysin-IR suggests multiple regulation sites for producing and maintaining pre-synaptic elements in the brain. PMID- 8723131 TI - Androgen modulates N-methyl-D-aspartate-mediated depolarization in CA1 hippocampal pyramidal cells. AB - Previously, research elucidating steroid hormone actions in the central nervous system has focused on their role in sexual reproduction and maintaining homeostasis. The hippocampus is a target of steroid modulation and is involved in the development of emotional behavior and memory storage. Area CA1 of the hippocampus contains a high density of androgen receptor (AR) and N-methyl-D aspartate (NMDA) receptors. NMDA receptors underlie excitatory synaptic transmission and excitotoxicity in CA1 neurons. The effects of AR activation on the neurophysiology of hippocampal pyramidal neurons is unknown. Standard intracellular recording techniques in hippocampal slices were used to investigate the effects of the non-aromatizable androgen, 5-alpha-dihydrotestos-terone proprionate (DHTP), on CA1 pyramidal cell characteristics and NMDA receptor mediated responses. Male Sprague-Dawley rats were unoperated, sham-operated (SHAM), gonadectomized (GDX), or gonadectomized with DHTP replacement therapy (GDX + DHTP). Neuronal AR was saturated by DHTP treatment as determined by binding studies and immunocytochemistry. Chronic DHTP treatment increased the action potential duration and decreased the fast afterhyperpolarization (fAHP) amplitude. To test the effect of DHTP on glutamate receptor-mediated responses, hippocampal slices were exposed to increasing concentrations of NMDA. In pyramidal cells from SHAM and GDX-treated animals, 30 microM NMDA induced an irreversible depolarization; the membrane potential of pyramidal cells from GDX + DHTP-treated animals recovered to baseline. The effect of DHTP was time dependent, implicating protein synthetic mechanisms. Our findings demonstrate that androgens can influence pyramidal cell characteristics and neurotransmitter evoked actions in hippocampal CA1 pyramidal cells. PMID- 8723132 TI - Regulation of serotonin-facilitated dopamine release in vivo: the role of protein kinase A activating transduction mechanisms. AB - Recent neuroanatomical, biochemical, and electrophysiological studies suggest that serotonin (5HT) can modulate dopaminergic function at the level of the cell body and the nerve terminal. The receptor subtypes, regulatory processes, and intracellular transduction mechanisms mediating these interactions remain to be characterized. The potential involvement of cAMP in mediating 5HT-facilitated increases in extracellular levels of striatal dopamine (DA) was assessed using in vivo microdialysis. Local infusion of 0.4 nmol 5HT delivered via probes located in the anterior striata of chloral hydrate-anesthetized male rats significantly increased extracellular DA levels to approximately 700% of basal control levels. Local, intrastriatal infusion of either 2 nmol forskolin, 2 nmol rolipram, 100 nmol isobutylmethylxanthine, or 200 nmol dibutyryl cAMP significantly increased basal DA levels to 28 +/- 3%, 143 +/- 5%, 56 +/- 7%, and 52 +/- 3% above control levels, respectively. Additionally, coperfusion of any of these agents with 5HT significantly decreased the 5HT-facilitory effect on DA release to approximately 50% of observed 5HT controls. The current results suggest a role for the cAMP second-messenger systems in modulating 5HT-facilitated DA release. PMID- 8723133 TI - Noradrenergic mechanisms in stress and anxiety: I. Preclinical studies. AB - There is considerable preclinical evidence for a relationship between noradrenergic brain systems and behaviors associated with stress and anxiety. The majority of noradrenergic neurons are located in the locus coeruleus (pons), with projections throughout the cerebral cortex and multiple subcortical areas, including hippocampus, amygdala, thalamus, and hypothalamus. This neuroanatomical formation of the noradrenergic system makes it well suited to rapidly and globally modulate brain function in response to changes in the environment, as occurs during the presentation of stressors. Stress exposure is associated with an increase in firing of the locus coeruleus and with associated increased release and turnover of norepinephrine in brain regions which receive noradrenergic innervation. Increased firing of the locus coeruleus is also associated with behavioral manifestations of fear, such as arched back and piloerection in the cat. Exposure to chronic stress results in long-term alterations in locus coeruleus firing and norepinephrine release in target brain regions of the locus coeruleus. Norepinephrine is also involved in neural mechanisms such as sensitization and fear conditioning, which are associated with stress. These findings are relevant to an understanding of psychiatric disorders, such as panic disorder and post-traumatic stress disorder (PTSD), the symptoms of which have been hypothesized to be related to alterations in noradrenergic function. PMID- 8723134 TI - Noradrenergic mechanisms in stress and anxiety: II. Clinical studies. AB - Studies in animals have shown a relationship between alterations in noradrenergic brain system function and behaviors of anxiety and fear. These findings have generated the hypothesis that the symptoms seen in patients with anxiety disorders may be related to alterations in noradrenergic function. A number of clinical studies have tested this hypothesis, utilizing measures of catecholaminergic function such as heart rate and blood pressure, measurement of norepinephrine and its metabolites in urine and plasma and adrenergic receptor binding in platelets, as well as pharmacological challenge to the noradrenergic system. Acute stressors, such as public speaking, have been associated with an increase in heart rate, blood pressure, and norepinephrine and its metabolites in urine and plasma. Findings in patients with panic disorder at baseline related to heart rate, blood pressure, baseline norepinephrine and its metabolites, and platelet adrenergic receptors have been mixed, while the most consistent findings have been blunted growth hormone response to clonidine and increased 3-methoxy-4 hydroxy-phenylethylene-glucol (MHPG) and anxiety following stimulation of the noradrenergic system with yohimbine. Baseline measures of noradrenergic function in patients with posttraumatic stress disorder (PTSD) have also been mixed, while an increased heart, blood pressure and norepinephrine response to traumatic reminders, as well as increased behavioral (as well as different brain metabolic) response to yohimbine, have been found in PTSD. There are fewer studies of noradrenergic function in the other anxiety disorders, and the findings there have not been consistent. These studies provide evidence for increased noradrenergic responsiveness in panic disorder and PTSD, although there does not appear to be an alteration in baseline noradrenergic function in these patients. PMID- 8723135 TI - Examination of the action of 3,4-methylenedioxymethamphetamine on rat A10 dopamine neurons. AB - Extracellular single cell recording was used to examine the effect of intravenous administration of (-), (+), and (+/-)-3,4-methylenedioxymethamphetamine (MDMA) on A10 dopamine (DA) neurons in chloral hydrate anesthetized male rats. Both (+/-) MDMA and (+)-MDMA inhibited the firing rate of most (79%) A10 DA cells. By contrast, (-)-MDMA induced either no effect or a slight increase in the firing rate of these cells. Analysis of the effects of (+/-)-MDMA on the firing pattern of the DA cells revealed on overall decrease in the percentage of spikes in bursts but both increases and decreases were seen in the coefficient of variation of interspike intervals. To determine the contribution of 5-HT and DA to the (+/ )-MDMA-induced inhibition of A10 DA cells rats were pretreated either with the 5 HT synthesis inhibitor p-chlorophenylalanine (PCPA) or the DA synthesis inhibitor alpha-methyl-p-tyrosine (AMPT). Pretreatment of rats with PCPA did not reduce the ability of (+/-)-MDMA to inhibit the DA cells. However, in rats pretreated with AMPT, the (+/-)-MDMA-induced inhibition was blocked and some cells (44%) showed instead an increase in firing rate following administration of (+/-)-MDMA. The administration of l-3,4-dihydroxyphenylalanine (L-DOPA) to AMPT-treated rats rapidly restored the inhibition of cell firing by (+/-)-MDMA. In conclusion, the results reported here demonstrate that MDMA has an overall inhibitory effect on A10 DA cells. Despite MDMA's greater potency in releasing 5-HT compared to DA, the inhibitory effect of this drug on A10 DA cells appears to be mediated by the latter transmitter. PMID- 8723136 TI - S-nitrosothiols and nitric oxide, but not sodium nitroprusside, protect nigrostriatal dopamine neurons against iron-induced oxidative stress in vivo. AB - Intranigral infusion of ferrous citrate (4.2 nmol) induced an acute lipid peroxidation in the substantia nigra and a chronic dopamine depletion in the striatum of rat nigrostriatal system. Coinfusion of 8.4 nmol nitric oxide donors such as S-nitrosoglutathione (GSNO) and S-nitroso-N-acetylpenicillamine (SNAP) or nitric oxide (approximately 2 nmol) protected nigrostriatal neurons against iron induced lipid peroxidation and associated oxidative injury. However, sodium nitroprusside (SNP, 8.4 nmol) augmented dopamine depletion caused by ferrous citrate because SNP is a ferricyanide complex. The present in vivo results indicate that nitric oxide and S-nitrosothiols are antioxidants which can protect brain dopamine neurons against oxidant stress/damage. PMID- 8723137 TI - Abnormal involuntary movements induced by subthalamic nucleus stimulation in parkinsonian patients. AB - Chronic electrical subthalamic nucleus (STN) stimulation is under investigation for alleviating parkinsonian symptoms. STN alterations may carry the risk of provoking abnormal involuntary movements (AIMs). We took advantage of the reversibility of the stimulation technique to assess the possibility of inducing AIMs, using different electrical variables with or without concomitant levodopa intake. Above a given threshold voltage, stimulation could induce contralateral distal mobile AIMs or hemiballism in the off-drug condition in two patients. AIMs occurred after a latency that varied from a few minutes up to several hours after switching on the stimulator. Hemiballism immediately disappeared upon switching off the stimulator. In these patients, levodopa had never provoked that type of AIMs before surgery. Levodopa-induced AIMs were not modified by electrical stimulation, but off-phase dystonia disappeared in one patient. Stimulation of the STN induced AIMs that resembled both those observed following spontaneous lesions of the STN and levodopa-induced diphasic AIMs in parkinsonian patients. As electrical stimulation provoked AIMs and antiparkinsonian benefit occurred with different electrical variables and different timing, the mechanisms responsible for these two effects could be distinct. PMID- 8723138 TI - Clinical course of patients with idiopathic Parkinson's disease. AB - We determined the variables associated with the progression and duration of illness of patients with Parkinson's disease (PD) and investigated the cause of death. In 474 patients with parkinsonism, who visited the Academic Hospital between January 1, 1960 and August 31, 1993, we did a survival analysis with the following covariates: age at onset, initial symptom (tremor or rigidity/hypokinesia), age at reaching Hoehn and Yahr stage III, dementia-free period, and levodopa treatment. A total of 345 patients with parkinsonism fulfilled the criteria of idiopathic PD; 258 of them were still alive on the closing date of this study. There were significantly more men than women (1.43:1). Medical advice was sought in an earlier stage by men and by patients with tremor as presenting symptom. For patients with rigidity/hypokinesia as first symptom, the duration of illness until reaching Hoehn and Yahr stage III was shorter than for patients starting with tremor alone. If the initial symptom is tremor, patients develop dementia less frequently and later after onset than when tremor is not involved. Reaching Hoehn and Yahr stage III and developing dementia both, limit the patient's survival time. The mortality risk for a patient was found to be increased from the moment levodopa treatment was started as compared to those patients who had not yet started with the treatment. The effect of levodopa on survival could not be disentangled from effects of other factors related to the start of levodopa treatment. PMID- 8723139 TI - Basal ganglia iron content in Parkinson's disease measured with magnetic resonance. AB - The possibility of using magnetic resonance (MR) to evaluate the severity of the pathological changes of Parkinson's disease (PD) is suggested by the known accumulation of iron in the basal ganglia in PD and the reduced signal evident from this area with conventional T2-weighted MR imaging. To improve the specificity of MR for the measurement of tissue iron content, we have developed a method that quantifies the effects of paramagnetic centers sequestered inside cellular membranes, based on the echo time dependence of the decay of transverse magnetization caused by the local field inhomogeneities which are due to intracellular iron. This method enables an index of local tissue iron content to be calculated for structures of the basal ganglia. We report here the application of this method to a series of patients with PD (n = 12) and of normal, age matched controls (n = 13). Our objective was to determine whether this measurement of basal ganglia iron concentration correlates with the presence and severity of PD. We observed a significant increase in iron content in both the putamen and pallidum in PD as well as a correlation with the severity of clinical symptomatology. More severely affected patients had a higher iron content in both of these structures. Our results suggest that this MR measurement may provide a noninvasive method of measuring the severity of the pathological changes underlying PD. PMID- 8723140 TI - A randomized, double-blind, placebo-controlled study to evaluate botulinum toxin type A in essential hand tremor. AB - Twenty-five patients with hand tremor of 2+ (moderate) to 4+ (severe) on the tremor severity rating scale were randomized to receive either 50 U of botulinum toxin (BTX) type A (Allergan, Irvine, CA) or placebo injections into the wrist flexors and extensors of the dominant limb. If patients failed to respond to the initial injection, they were eligible to receive another injection of 100 U 4 weeks later. Rest, postural, and kinetic tremors were evaluated at 2- to 4-week intervals over a 16-week study period using tremor severity rating scales, accelerometry, and assessments of improvement and disability. A significant improvement (p < 0.05) was observed on the tremor severity rating scale 4 weeks after injection in patients treated with BTX as compared with placebo, and this effect was maintained for the duration of the study. Four weeks after injection, 75% of BTX-treated patients vs. 27% of placebo-treated patients (p < 0.05) reported mild to moderate improvement (peak effect rating > or = 2). There were no significant improvements in functional rating scales, although trends were observed for some items. Postural accelerometry measurements showed a > or = 30% reduction in amplitude in nine of 12 BTX-treated subjects and in one of nine placebo-treated subjects (p < 0.05). Although all patients treated with BTX reported some degree of finger weakness, no severe, irreversible, or unexpected adverse events occurred. Chemodenervation with BTX may significantly ameliorate essential hand tremor in patients who fail to improve with conventional pharmacologic therapy. PMID- 8723141 TI - Effect of the putative dopamine D1 agonist and D2 antagonist FCE 23884 on Parkinson's disease. AB - The ergoline derivative FCE 23,884 acts as a dopamine D1 agonist in untreated parkinsonian animals and as a D2 antagonist in animals whose dopamine system is intact or levodopa treated. To evaluate whether this dual action might benefit patients with Parkinson's disease (PD) who have developed levodopa-induced dyskinesias, the motor effects of FCE 23,884 were examined in seven such individuals using a double-blind, placebo-controlled design. At doses up to the maximum tolerated dose (3.5 +/- 0.5 mg), FCE 23,884 monotherapy did not affect parkinsonian severity. On the other hand, coadministration of FCE 23,884 with a mildly dyskinetic dose of levodopa, infused intravenously under steady-state conditions, reduced the antiparkinson response by 54 +/- 19% and tended to diminish dyskinesia severity. The results thus fail to suggest any useful role for FCE 23,884 in the symptomatic treatment of PD. Although D2 receptor blockade provided by FCE 23,884 antagonizes both the antiparkinson and dyskinesigenic responses to levodopa, the degree of D1 receptor stimulation appears insufficient to ameliorate parkinsonian symptomatology. PMID- 8723143 TI - Sympathetic skin response and R-R interval variability in multiple system atrophy and idiopathic Parkinson's disease. AB - We compared autonomic function in patients with multiple system atrophy (MSA) or with idiopathic Parkinson's disease (IPD) by measuring sympathetic skin response (SSR) and R-R interval variability (RRIV). SSR was investigated in 26 patients (13 with MSA and 13 patients with IPD). RRIV during deep breathing, Valsalva maneuver, and on standing was investigated in 20 patients (nine with MSA and 11 with IPD). MSA and IPD patients had similar age, illness duration, and therapy. Abnormal SSR was more frequent in MSA (69%) than in IPD (7.7%; x2, 10.4; p < 0.002). RRIV during deep breathing and the Valsalva maneuver was lower in MSA than in IPD (p = 0.02). RRIV during standing up was not significantly different in IPD and MSA. These differences between MSA and IPD may be due to more severe and widespread autonomic disturbance in MSA, related to more severe neuropathologic involvement of the autonomic nervous system. SSR and RRIV may aid in the differential diagnosis of parkinsonism and help to exclude from clinical trials MSA patients clinically misdiagnosed as having IPD. PMID- 8723142 TI - Neutrophil function, nitric oxide, and blood oxidative stress in Parkinson's disease. AB - We studied nitrogen radical nitric oxide (.NO) release and reactive oxygen species (ROS) production by isolated neutrophils after phorbol myristate acetate (PMA) stimulation in 12 newly diagnosed and nine treated Parkinson's disease (PD) patients and 10 age-matched healthy controls. Neutrophils of both groups of PD patients had an elevated PMA-activated release of .NO [61 and 57%, respectively, higher than that of controls (p < 0.05)]. In contrast, H2O2 release was only significantly increased by 56% in chronically treated patients. In agreement, the maximum rate of luminol-dependent chemiluminescence, which partly represents O2- H2O2- .NO interactions, was increased only in the treated group. When other blood markers of oxidative stress were compared, only erythrocyte catalase activity was decreased in both PD patient series by 33 and 39%, respectively (p < 0.05), whereas plasma antioxidant capacity and erythrocyte superoxide dismutase activity levels were decreased only in treated PD patients. This study suggests that neutrophils express a primary alteration of .NO release in PD patients, whereas H2O2 and oxidative-stress parameters are more probably related to the evolution of PD or to effects of treatment with L-dopa. PMID- 8723144 TI - A controlled trial of remacemide hydrochloride in Huntington's disease. AB - We conducted a randomized, double-blind, placebo-controlled tolerability study of a N-methyl-D-aspartate (NMDA) glutamate receptor ion-channel blocker, remacemide hydrochloride, in 31 independently ambulatory patients (18 men, 13 women) with Huntington's disease (HD). Subjects were randomized to receive either placebo or active remacemide at dosages of 200 mg/day or 600 mg/day. The primary outcome measure was the proportion of subjects able to complete the study with the assigned treatment. Remacemide was generally well tolerated, and no significant differences between the treatment arms were found in the primary outcome measure. A trend toward improvement in chorea was observed among subjects administered remacemide 200 mg/day. Based on the tolerability and safety demonstrated during this short-term trial, remacemide warrants more extended controlled investigation in patients with HD. PMID- 8723145 TI - Stuttering may be a type of action dystonia. AB - We observed abnormal involuntary movements, involving principally the facial and neck muscles, in 23 patients with stuttering. These movements were similar to involuntary movements seen in distinct dystonic syndromes. There was a history of stuttering in the first degree relatives of six patients. The association of stuttering with degenerative neurologic disorders and focal brain lesions, cerebral blood flow changes in patients with developmental stuttering, its occurrence as a side effect of centrally acting drugs, induction and alleviation of stuttering by mechanical perturbation, or by electrical stimulation of the thalamus, a strong genetic predisposition with male preponderance, and the statistically significant occurrence of stuttering in the family history of patients with idiopathic torsion dystonia suggest an organic basis for developmental stuttering. These findings and the reported similarities between the involuntary movements associated with stuttering and dystonic involuntary movements support the hypothesis that stuttering is a form of segmental or focal action dystonia. PMID- 8723146 TI - The electroencephalogram in acetazolamide-responsive periodic ataxia. AB - Acetazolamide-responsive periodic ataxia (ARPA) is a rare movement disorder, characterized by recurrent episodes of vertigo, cerebellar ataxia, and nystagmus, which has recently been characterized genetically. The pathophysiology is unknown, but it is probably not epileptic. By definition, acetazolamide produces an impressive symptomatic relief. Because of the paroxysmal nature of the disorder, EEG tracings were often obtained. We report four new cases (two familial and two sporadic) with typical ARPA (none of whom had metabolic abnormalities or continuous electrical muscle activity) and review the EEG findings associated with this disorder. EEG findings were reported in 18 kindreds and nine sporadic cases (including ours). EEG was described in 54 of the 140 affected cases and was abnormal in 52% (28/54). Most commonly seen was intermittent generalized slow activity, observed in 35% (19/54), frequently intermingled with spikes (10 cases). Other abnormalities included nonspecific mild generalized or focal slowing in seven (13%) and focal epileptic activity in two (4%) patients. The paroxysmal EEG activity frequently seen in ARPA should not establish a diagnosis of epilepsy. Although not specific, it may suggest the correct diagnosis and indicate treatment with acetazolamide. PMID- 8723147 TI - Computational analysis of open loop handwriting movements in Parkinson's disease: a rapid method to detect dopamimetic effects. AB - We used a computational analysis of open loop handwriting movements and a clinical rating scale for monitoring the effect of apomorphine in 16 patients with early untreated parkinsonism [subsequently L-DOPA responsive, probable Parkinson's disease (PD)], six patients with long-standing PD with L-DOPA associated motor fluctuations, and seven patients with known L-DOPA unresponsive parkinsonism. Subjects were instructed to write fluently concentric circles of approximately 12 mm in diameter. Movements were recorded for two periods of 3 s each, using a digitizing tablet. Mean peak velocity (Vmax) and mean peak acceleration (Amax) were determined. In addition, two sensitive indices describing the degree of automation of handwriting were derived: (a) NCV, calculated as the mean Number of Changes in direction of Velocity per half circle, and (b) NCA, the mean Number of Changes in the direction of Acceleration. Clinical rating was performed according to the Unified Parkinson's Disease Rating Scale part III (UPDRS III). After apomorphine injection, the patients with early untreated probable PD showed significant improvement of Vmax, Amax, NCV, NCA, and UPDRS III scores. Likewise, the patients with long-standing PD improved significantly in all kinematic parameters and UPDRS III scores. Patients with L DOPA unresponsive parkinsonism failed to change significantly in any of the parameters tested. These observations suggest that the computer-assisted analysis of automated handwriting movements can be used as an objective quick method for quantifying dopamimetic effects on the kinematics of handwriting movements in parkinsonian patients. PMID- 8723148 TI - Case 1, 1996: rapidly progressive parkinsonism, incontinence, impotency, and levodopa-induced moaning in a patient with multiple myeloma. PMID- 8723149 TI - Effect of stimulation in the ventral intermediate nucleus of the thalamus on limb control in Parkinson's disease: a case study. AB - This study focuses on upper extremity strength and movement control in a patient with Parkinson's disease who had stimulating electrodes surgically implanted in the ventral intermediate nucleus (VIM) of the left thalamus. We examined torque generation and control of movement distance in single degree-of-freedom elbow movements under three different stimulation conditions: (a) no stimulation, (b) high stimulation, in which tremor was minimized but there was also tingling and perceived weakness, and (c) moderate stimulation, in which tremor was partially reduced, but there was also a subjective sense of increased strength compared with the high-stimulation condition. The patient's ability to generate both steady torque and rapid movements was poorest with no stimulation. The patient generated the largest torques with moderate stimulation and performed the fastest movements with high stimulation. However, even with tremor minimized, the patient's electromyogram (EMG) burst patterns were not typical of those of neurologically healthy subjects, although the movements were clearly improved. PMID- 8723150 TI - Familial paroxysmal dystonic choreoathetosis revisited. AB - A case of familial paroxysmal dystonic choreoathetosis (PDC) documented by video/EEG monitoring is described. The father of the proband is affected by exertional cramping but not PDC, lending support to the previous hypothesis that exertional cramping may represent a "forme fruste" or the incomplete expression of PDC. Other family members affected by PDC are women, with exercise-induced cramping alone found in two men. Two of the women report prolonged exertion as a precipitant of lengthy spells consistent with typical PDC rather than the previously described "intermediate," exercise-induced form of PDC. Exertional cramping in families affected by PDC may represent the variable expression of the "dystonia gene" in male members. Conversely, exercise-induced PDC, both of the intermediate and longer form described here, may have a predilection to manifest in women. PMID- 8723151 TI - Assessment of coenzyme Q10 tolerability in Huntington's disease. AB - We performed a 6-month open-label trial to evaluate the tolerability and efficacy of coenzyme Q10 (CoQ) in 10 patients with Huntington's disease (HD). Subjects were evaluated at baseline, 3 months, and 6 months using the HD Rating Scale (HDRS), the HD Functional Capacity Scale (HDFCS), and standardized neuropsychological measures. Adverse events (AEs) were assessed by telephone interview every month. CoQ doses ranged from 600 to 1,200 mg per day. All subjects completed the study, although four subjects reported mild AEs, including headache, heartburn, fatigue, and increased involuntary movements. There was no significant effect of the treatment on the clinical ratings. The good tolerability of CoQ suggests that it is a good candidate for evaluation in long term clinical trials designed to slow the progression of HD. PMID- 8723152 TI - Complex movement disorders induced by fluoxetine. AB - We report two cases of complex movement disorders induced by fluoxetine. A 72 year-old woman developed rhythmic palatal movements, myoclonus, chorea, and possibly dystonia after 2 years of therapy with fluoxetine. On withdrawal of fluoxetine, the movements abated after 5 days and did not recur. The second patient, a 58-year-old man, developed myoclonic jerking and rapid, stereotypic movements of his toes after a year of fluoxetine therapy. These complex movements have not been reported previously as an adverse effect of fluoxetine. PMID- 8723153 TI - High-dose pergolide monotherapy in the treatment of severe levodopa-induced dyskinesias. PMID- 8723154 TI - Supranuclear palsy of voluntary eyelid closure: electrophysiological correlate. PMID- 8723155 TI - A case of pelvic floor myoclonic jerk syndrome. PMID- 8723156 TI - Mycoplasma pneumoniae postinfectious encephalomyelitis with bilateral striatal necrosis. PMID- 8723157 TI - Cervical dystonia due to a frontal meningioma. PMID- 8723158 TI - No effects on heart-rate variability and cardiovascular reflex tests after botulinum toxin treatment of cervical dystonia. PMID- 8723159 TI - Painful calf, moving foot. PMID- 8723160 TI - Hemidystonia occurring in a patient with sarcoidosis. PMID- 8723161 TI - Prevalence of focal dystonias in the western area of Tottori Prefecture in Japan. PMID- 8723162 TI - Does levodopa therapy delay death in Parkinson's disease? PMID- 8723163 TI - Teaching tape for the motor section of the Unified Parkinson's Disease Rating Scale. PMID- 8723165 TI - Usefulness of diltiazem in the acute management of supraventricular tachyarrhythmias in the elderly. AB - Acute management of supraventricular tachyarrhythmias is often difficult in elderly patients. Diltiazem was given intravenously (loading dose of 0.25 mg/kg over 2 minutes followed by a 4 mg/kg/24 hr infusion) in 37 elderly patients (mean age 70 years, range 60-91). Fifteen out of the 37 patients (41%) had left ventricular cardiac disease, 12 (32%) had cor pulmonale, and 10 (27%) had no obvious cardiac disease. Hemodynamic tolerance of the supraventricular tachyarrhythmia was poor in 12 patients. A good result was defined as a return to sinus rhythm after bolus or infusion, or as a slowing of the ventricular rate (VR) to less than 100 beats/min. Of the 23 patients in atrial fibrillation, about half reverted to sinus rhythm after diltiazem, and in most of the others the ventricular rate decreased to less than 100 beats/min. Side effects occurred in 10 patients (bradycardia in 6, cutaneous rash in 2, hypotension in 2). They rapidly reversed after cessation of diltiazem. They were responsible for 2 out of the 5 poor results. Thus, diltiazem appeared effective and safe when used carefully in elderly patients with supraventricular tachyarrhythmia. PMID- 8723164 TI - Intermittent or continuous transdermal nitroglycerin: still an issue, or is the case closed? AB - After a decade of controversy and debate, many experts have now concluded that continuous nitroglycerin patch treatment leads to complete tolerance development and therefore cannot be recommended for any angina patient. This conclusion is largely based on the disappointing results of the large Transdermal Nitroglycerin Cooperative Study, in which continuous patch treatment in doses of 15-105 mg daily failed to increase exercise duration more than placebo after 2 and 8 weeks of treatment. However, other well-designed studies recently reported maintained efficacy during continuous treatment, and the differences in results has remained unexplained. The disagreeing data may be better understood if certain facts are considered: (1) The cooperative study tested a patient population with a very low first-dose treatment response--only 34 seconds (or 10-12%) improvement compared with placebo. At the end of the study, 25% of the patients terminated exercise for reasons other than angina, and a reduced nitrate responsiveness had developed, even in the placebo group. (2) Patients who demonstrate a large first dose nitrate responsiveness tend to be less susceptible to tolerance development. (3) Even during continuous therapy, maintained efficacy is often observed in exercise tests done 2-5 hours after patch renewal, while typically no effect is seen at the end of the application period. Attenuation of the initial effects is seen with all long-acting nitrate treatment regimens, but the degree of tolerance varies with the patient population, the efficacy parameter (exercise test vs. attack counts), the timing of the efficacy test, the patch dose, and whether or not nitrate-free (-low) intervals are used. In general, intermittent patch therapy is superior to continuous therapy in improving exercise duration, but even continuous therapy may retain some effect. Rebound phenomena do occur but are clinically relevant only in a minority of the patients. Rebound phenomena are not a problem during continuous therapy, which therefore may be of value in patients with frequent and/or nocturnal angina attacks. Patients experiencing angina during exercise only and with low first-dose effects are likely to benefit more from intermittent therapy. Doses of 0.6-0.8 mg/hr (15-20 mg/24 hr) are usually optimal, and the efficacy is comparable with that observed after oral nitrates. PMID- 8723166 TI - Coronary vasomotor responses in cyclosporine-treated piglets. AB - Chronic treatment with cyclosporine A (Cx) seems to produce a decreased ability of the endothelium to secrete nitric oxide. However, its effect on the coronary arterial system remains controversial. Therefore, coronary arteries isolated from piglets treated by intramuscular injections of Cx (10 mg/kg/day, IM, for 22 days) were studied in organ baths and compared with those isolated from control animals (IM injections of the Cx solvent). Depolarization-induced contractions (KCl 120 mM) were similar in both groups, whereas the acetylcholine-induced contractions (percent of 120 mM KCl) were enhanced: The area under the curve (AUC) was 245 +/- 51 in the Cx group versus 110 +/- 15 in the control group (p < 0.05). Removal of the endothelium did not significantly modify the acetylcholine-induced contractions in both groups and, therefore, did not attenuate the enhanced responsiveness to acetylcholine in the Cx group. On unrubbed rings contracted with prostaglandin F2 alpha, the endothelium-dependent relaxations from serotonin (in the presence of 1 microM ketanserin) were reduced: The AUC was 479 +/- 24 in the Cx group versus 385 +/- 22 in the control group (p < 0.02). Larger AUC values were also found for bradykinin and substance P in the Cx group: 158 +/- 18 versus 55 +/- 17 (in the control group, p < 0.01) and 198 +/- 8 versus 145 +/- 12 (p < 0.01), respectively. Nevertheless, no alteration in calcium ionophore-induced relaxations was observed: The AUC was 217 +/- 10 in the Cx group and 224 +/- 18 in the control group (NS). Indomethacin incubation (10 microM) did not prevent the impairment in endothelium-dependent relaxations and did not attenuate the cyclosporine-induced augmentation of acetylcholine-induced contractions. Thus, chronic administration of cyclosporine to piglets impairs the coronary endothelial function and produces functional changes in smooth muscle cells. These alterations may play a role in the occurrence of cardiac graft vasculopathy. PMID- 8723167 TI - Tilisolol hydrochloride dilates coronary arteries through an ATP-sensitive K(+) channel opening mechanism in dogs. AB - Tilisolol is a beta-blocking agent with vasodilatory properties that was recently shown to possess a potassium (K+) channel opening activity. We investigated whether tilisolol has vasodilatory effects on coronary circulation in dogs. Mongrel dogs were chronically instrumented for measurements of circumflex coronary artery diameter (CoD) and coronary blood flow (CBF). We compared the effects of tilisolol on dog coronary arteries with those of two beta-blockers, propranolol and arotinolol. Both propranolol (1 mg/kg, intravenously, i.v.) and arotinolol (0.25 mg/kg, i.v.) decreased CoD and increased coronary vascular resistance (CVR). Tilisolol (2 mg/kg, i.v.) decreased CVR but had no significant effect on CoD. To investigate the mechanism of the coronary action of tilisolol, we examined differences in the response to tilisolol in the absence and presence of glibenclamide, an ATP-sensitive K+ channel blocker. Tilisolol (1,2,4, and 8 mg/kg, i.v.) produced a dose-dependent decrease in CVR without glibenclamide, whereas pretreatment with glibenclamide significantly suppressed this effect. Without glibenclamide, tilisolol had no significant effect on CoD at doses of 1-4 mg/kg (i.v.). However, at the higher dose of 8 mg/kg (i.v.), tilisolol significantly increased CoD (1.00 +/- 0.15%, p < 0.01). After pretreatment with glibenclamide, tilisolol (1-8 mg/kg, i.v.) produced a significant decrease in CoD. Therefore, we concluded that tilisolol exerts its vasodilatory effect on the coronary circulation through an ATP-sensitive K+ channel opening mechanism, and that its vasodilatory action is more prominent in coronary resistance vessels than in large coronary arteries. PMID- 8723168 TI - A novel catecholamine, arbutamine, for a pharmacological cardiac stress agent. AB - Arbutamine, developed for use as a cardiac stress agent, was compared with isoproterenol and dobutamine in anesthetized dogs for cardiovascular actions prior to and after beta-adrenergic blockade with propranolol. The efficacy and safety of arbutamine were also evaluated in a canine model of myocardial ischemia obtained by partially occluding the left anterior descending coronary artery. Comparison of hemodynamic variables in normal dogs showed that arbutamine was approximately equipotent to isoproterenol in increasing heart rate and cardiac contractility, and in decreasing total peripheral vascular resistance and mean arterial blood pressure. Arbutamine was 210 times more potent than dobutamine in increasing cardiac contractility by 70%; however, at this dose dobutamine exhibited a negative chronotropic response. Beta-adrenergic blockade with propranolol shifted the agonist's dose-response curves for heart rate and contractility to the right; however, low doses of dobutamine exhibited a negative chronotropic effect and increased the total peripheral vascular resistance. In dogs subjected to partial left anterior descending coronary artery occlusion, arbutamine produced significant ST-segment deflections, beginning at a dose of 0.1 nmol/kg/min. Impairment of segment shortening, reflecting cardiac wall motion abnormality, was evident at a dose of 0.3 nmol/kg/min. Isoproterenol did not cause significant changes in these parameters. These results show that arbutamine is capable of producing graded increments in cardiac contractility and rate before and after beta-adrenergic blockage in normal dogs. In dogs subjected to coronary artery occlusion, it is capable of provoking myocardial ischemia at dose levels devoid of toxicity. PMID- 8723169 TI - Characterization of the adrenergic activity of arbutamine, a novel agent for pharmacological stress testing. AB - In this study, we characterized the interactions of arbutamine, a novel catecholamine developed for use as a cardiac stress testing agent, with different adrenergic receptor subtypes in vitro. These effects were compared with those of isoproterenol. In the electrically stimulated left atria of rats, arbutamine increased contractile force. The pD2 values (- log of the dose that produces 50% of the maximal responses) for arbutamine and isoproterenol were 8.45 +/- 0.15 and 8.55 +/- 0.02, respectively. Metoprolol shifted the concentration-effect curves for both isoproterenol and arbutamine to the right with a pA2 value (- log of the dose of the antagonist that reduces the maximal responses of an agonist to 50%) of 7.22-7.5. Both arbutamine and isoproterenol increased the rate of spontaneously beating rat right atria with pD2 values of 9.0 +/- 0.19 and 8.82 +/ 0.18, respectively. The affinity constants (KA) of arbutamine and isoproterenol for cardiac beta1-adrenergic receptors, as determined by competition binding assays, were found to be 7.32 and 6.04, respectively. In guinea pig trachea, arbutamine and isoproterenol produced a concentration-dependent relaxation that was blocked by propranolol. Their pD2 values were 7.9 +/- 0.1 and 8.2 +/- 0.1, respectively. Arbutamine contracted isolated rat aortic rings with a maximal increase of 38.1 +/- 6.7% that of 10 microM of norepinephrine. In rat white adipocytes, arbutamine, isoproterenol, and BRL-37344 stimulated glycerol release, with the order of potency being BRL-37344 > arbutamine > isoproterenol. In hamster brown adipocytes, the order was arbutamine > isoproterenol > BRL-37344. Moreover, arbutamine stimulated beta3-adrenergic receptors in guinea pig ileum. In conclusion, arbutamine is a novel catecholamine with similar potency and efficacy to that of isoproterenol. It stimulates cardiac beta1-, tracheal beta2-, and adiopocyte beta3-adrenergic receptors. Arbutamine does not stimulate alpha adrenergic receptors at concentrations that were high enough to maximally activate the beta-adrenergic receptors. PMID- 8723171 TI - Pranidipine, a novel calcium antagonist, once daily, for the treatment of hypertension: a multicenter, double-blind, placebo-controlled dose-finding study. AB - The antihypertensive effects and tolerance of once-daily (od), pranidipine, a novel dihydropyridine derivative with a long duration of action, were evaluated in a double-blind, placebo-controlled, parallel-group dose-finding study. A total of 199 patients, with a diastolic blood pressure (BP) of 95-115 mmHg, were included in the trial. After 4 weeks on placebo, patients were randomly assigned to either placebo or pranidipine at 1, 2, 4, or 8 mg od for a further 4 weeks. A dose response was seen in the reduction (delta) of diastolic BP: placebo, delta 1.7 mm Hg; 1 mg, delta 6.4 mmHg; 2 mg, delta 7.5 mmHg, p < 0.01; 4 mg, delta 11.5 mmHg, p < 0.01; and 8 mg, delta 10.6 mmHg, p < 0.01. There were no meaningful changes in heart rate. The number of responders (decrease of diastolic blood pressure to < 90 mmHg and by 10 mmHg or more from baseline value) in each group also revealed a dose-response relationship: placebo = 9%; 1 mg = 25%, n.s.; 2 mg = 27%, n.s.; 4 mg = 41.5%, p < 0.01; and 8 mg = 41%, p < 0.01 (compared with placebo). Plasma concentrations of pranidipine also demonstrated linear dose response relationships. An increase in adverse events was observed within the 8 mg group. The degree of reduction in BP and the number of responders were not greater in the 8 mg group compared with the 4 mg group, although the plasma concentration (mean values, ng/dl) of pranidine in the 8 mg group was higher (2.2 on day 42; 2.3 on day 56) compared with the 4 mg group (1.4 on day 42; 1.6 on day 56). In conclusion, pranidipine is a well-tolerated and 24-hour effective novel calcium antagonist that reduces BP in a dose-related manner up to 4 mg od. PMID- 8723170 TI - Influence of antihypertensive therapy with cilazapril and hydrochlorothiazide on the stiffness of the aorta. AB - The purpose of this study was to examine the effects of the angiotensin converting enzyme (ACE) inhibitor cilazapril on the elastic properties of the aorta. A standard diuretic antihypertensive drug, hydrochlorothiazide, served for comparisons. Increased aortic stiffness leads to a reduction of the buffering windkessel function and is a major component in the pathophysiology of systolic hypertension, inducing an increase in left ventricular afterload and arterial pulsatile stress as well as a decrease in the subendocardial blood supply. Stiffness of arteries increases with age and blood pressure, and depends on the functional elastic structures of the aortic wall. ACE inhibitors have been shown to directly influence elastic properties of peripheral arteries. Seventeen patients with mild to moderate essential hypertension (age 45-67 years) were treated for 3 months double-blind randomized with either cilazapril (C) 5 mg daily (n = 9) or hydrochlorothiazide (HCTZ) 25 mg daily (n = 8). Aortic elastic properties were noninvasively assessed by measurement of pulse wave velocity along the aorta at rest and during isometric handgrip stress. Accelerated pulse wave velocity indicates elevated arterial stiffness and vice versa. A pressure standardized index of aortic cross-sectional distensibility (2 m) was calculated from arterial mean pressure and pulse wave velocity. Compared with pretreatment values, both therapies significantly reduced blood pressure and pulse wave velocity at rest (C: 9.4 +/- 0.9 vs. 7.7 +/- 0.7 m/sec; HcTZ: 8.9 +/- 0.3 vs. 7.8 +/- 0.4 m/sec; means +/- SEM p < 0.05). During isometric stress only C showed a significant decrease in pulse wave velocity (C: 11.3 +/- 0.8 vs. 9.1 +/- 0.8 m/sec; HCTZ: 9.9 +/- 0.5 vs. 9.0 +/- 0.5 m/sec; means +/- SEM p < 0.05). The index 2m at rest and during handgrip increased significantly (p < 0.05) after C but not after HCTZ. With cilazapril we obtained steeper slopes for the treatment induced reductions in blood pressure and pulse wave velocity for both rest and handgrip stress values. Correlation of the data at rest and during stress revealed a direct relationship between blood pressure and pulse wave velocity. HCTZ linearly extended the relation observed before treatment toward lower values of blood pressure and corresponding pulse wave velocity without changing the relation per se. Cilazapril, in contrast, moved the relation between these variables and decelerated the pulse wave velocities to a greater extent than would have been expected from the corresponding blood pressure reduction (delta approximately 1 m/sec). These results in patients with mild to moderate essential hypertension support the idea that ACE inhibitors, in addition to reducing blood pressure, may exert an additional hemodynamic effect in improving the elastic properties of the aorta. PMID- 8723172 TI - Effects of beta-blockers on HMG CoA reductase and LDL receptor activity in cultured human skin fibroblasts. AB - Previous reports, based on clinical trials and animal experiments, suggest that beta-blockers may be useful in the prevention of atherosclerosis. Betaxolol, a new beta1-selective blocker, was shown to decrease plasma total and LDL cholesterol levels or to have no adverse effect on those [1-4]. While many reports deal with metabolism of triglyceride and high density lipoprotein, fewer publications about cholesterol metabolism are currently available. To clarify the mechanism by which beta-blockers affect lipid metabolism, we examined the effects of beta-blockers on HMG CoA reductase and LDL receptor activity in cultured human skin fibroblasts. L-propranolol, a nonselective beta-blocker, increased HMG CoA reductase activity and decreased LDL receptor activity. However, d-propranolol had no major effects on HMG CoA reductase activity. These results suggest that beta-blockers act on HMG CoA reductase through the beta receptors. Beta1-blocking action should decrease HMG CoA reductase activity and increase LDL receptor activity. In fact, betaxolol, a beta1-selective blocker, decreased HMG CoA reductase activity and increased LDL receptor activity, but metoprolol had no major effect. We speculate that the discrepancy between betaxolol and metoprolol in the effect on HMG CoA reductase and the LDL receptor might be due to the difference of the extent of beta1-selectivity. We conclude that beta1-selective blockers are antihypertensive agents potentially valuable in the prevention of atherosclerosis. PMID- 8723173 TI - Prediction of renal impairment in elderly patients with congestive heart failure treated with captopril. AB - This study assessed the usefulness of the oral captopril test in the prediction of renal impairment among elderly patients with congestive heart failure (CHF). Forty-seven patients aged > or = 65 years with CHF (EF < 40%) participated in a prospective nonrandomized series. Blood samples for plasma renin activity (PRA) were drawn before and 60 minutes after 50 mg of oral captopril. Twenty-four hours later, captopril was administered (up to 75 mg/day over a 4 day period), and renal laboratory and clinical assessment were performed at baseline and for a 9 day period. In 7 of 47 patients (14.9%), deterioration of renal function was observed. During the captopril test, the PRA increased significantly after 1 hour in almost all patients and the mean blood pressure decreased from 99.2 +/- 14.6 mmHg to 92.2 +/- 13.7 mmHg (p < 0.001). All patients whose baseline PRA level was < 1.9 ng/ml/hr and whose stimulated PRA was < 3.2 ng/ml/hr maintained a stable renal function throughout the study period. Significant statistical correlation (p < 0.05) was found between the initial PRA, the changes in PRA or mean blood pressure during the captopril test, and the change in plasma creatinine and creatinine clearance in the entire group, and was even more evident in a subgroup of patients with an ejection fraction > or = 30%. All these correlations were not statistically significant in the patients with an ejection fraction < 30%. It is thus concluded that measurement of pretreatment PRA levels might be a useful laboratory tool for predicting the renal safety of captopril use in patients with CHF whose EF > or = 30%. PMID- 8723175 TI - Measuring defensive medicine using clinical scenario surveys. AB - For more than two decades, advocates of malpractice system reform have claimed that the most damaging and costly result of the U.S. medical malpractice system is the practice of defensive medicine, in which physicians order tests and procedures primarily because of fear of malpractice liability. In this article, we discuss the issues raised by different definitions of defensive medicine and propose a working definition to guide measurement of the concept. We also consider the strengths and weaknesses of available approaches for measuring defensive medicine. Finally, we describe an empirical approach to measuring defensive medicine using clinical scenario surveys. The results suggest that, if physicians actually practice as they say they would in these surveys, defensive medicine does exist, although not to the extent suggested by anecdotal evidence or direct physician surveys. The results also suggest that defensive medicine varies considerably across clinical situations. In all of the scenarios, many physicians chose aggressive patient management styles even though conservative management was considered medically acceptable by the expert panels that developed the scenarios. In most cases, medical indications, not malpractice concerns, motivated clinical choices. Our results highlight the limitations of surveys as a method of measuring the extent of defensive medicine. The implications of managed care and health care reform for defensive medicine are also discussed. PMID- 8723174 TI - Acute enoximone effect on systemic and renal hemodynamics in patients with heart failure. AB - Patients with heart failure generally show improvement in their clinical condition after enoximone infusion over the period of treatment; this effect cannot be ascribed only to the known hemodynamic action of this drug. Thirty-six patients (age range 44-82 years) with heart failure (NYHA class II-IV) underwent 48-hour enoximone infusion to study whether this prolonged improvement might depend on changes in systemic or renal hemodynamics or in neurohormonal balance. All patients underwent Swan-Ganz hemodynamic monitoring; renal plasma flow, glomerular filtration rate, plasma atrial natriuretic factor (ANF), and plasma renin activity (PRA) were all measured at baseline, at the peak of the enoximone action, and 48 hours after drug discontinuation. The main hemodynamic parameters were significantly improved during enoximone infusion and after drug discontinuation. The cardiac index basal value of 2.2 +/- 0.1 l/min/m2 increased to 3.1 +/- 0.1 l/min/m2 after 24-hour therapy (p < 0.01); similarly, pulmonary wedge pressure, mean pulmonary arterial pressure, and right atrial pressure decreased markedly (p < 0.01). Beneficial effects were also observed in renal hemodynamics; indeed, renal plasma flow (basal value 485 +/- 39 ml/min) increased significantly after 24-hour enoximone infusion (575 +/- 35 ml/min; p < 0.01), and this tendency was also observed 48 hours after drug discontinuation. No significant modifications were observed in plasma hormone data; however, the PRA plasma level had a tendency to decrease. We conclude that in patients with heart failure, enoximone infusion has a less marked effect on renal hemodynamics, but this is more lasting than systemic hemodynamic effects. The tendency of PRA to decrease (although not statistically significant), still detectable 2 days after treatment in the presence of steady high plasma ANF concentrations, may also contribute to the paradoxical longlasting benefit despite the short-lived improvement in systemic hemodynamics after brief cycles of enoximone infusion. PMID- 8723176 TI - Physicians' personal malpractice experiences are not related to defensive clinical practices. AB - Whether personal malpractice experience is part of a tort signal prompting physicians to practice defensively is unclear. To explore this issue further, we assessed how physicians' malpractice experiences affect clinical decision making. We surveyed 1,540 physicians from four specialty groups (cardiologists, surgeons, obstetrician-gynecologists, and internists) using specialty-specific clinical scenarios. Physicians were in active private practice, were covered by a single malpractice insurer for five or more years, and worked in an eastern state. The net response rate was 54 percent (835 of 1,540) but measurable bias, based on practice characteristics, was negligible. Physicians evaluated clinical scenarios that were designed to maximize potential for finding positive defensive practices (extra tests and procedures). Then they rated how various factors influenced their decisions and answered questions on practice attitudes. The study compared management and testing recommendations among physicians with varying levels of malpractice exposure, which we defined in three separate ways. Participants were unaware of the study hypotheses. Physicians with greater malpractice experience showed no systematic differences in initial management choice or subsequent test recommendations. For example, similar percentages of internists in the top and bottom claims rate quartiles admitted a patient with syncope (78 percent versus 73 percent; p = 42), discharged a patient with nonspecific chest pain (80 percent versus 80 percent; p = .88), and delayed surgery in a patient with nonspecific changes on a electrocardiograph (58 percent versus 68 percent; p = .18). Attitudes about malpractice also did not differ with varying malpractice experience. Personal malpractice experience is not a predominant factor in the tort signal that prompts physicians to engage in defensive practices, to the extent that such practices exist. PMID- 8723177 TI - The use of low-osmolar contrast agents: technological change and defensive medicine. AB - The escalating cost of medical care in the United States, especially in the past decade, has resulted in efforts to identify the factors contributing to rising costs. One factor often assumed to cause higher medical costs is the physician's fear of liability for not using the latest available technology. In this article, we report the results of a case study we conducted to better understand the relationship between the introduction and use of one particular technology, low osmolar contrast agents, and liability concerns. Our study suggests that both clinicians and administrators are primarily guided by the medical benefits of low osmolar contrast agents, and that liability concerns, although widespread, are of secondary importance. The inability to control this and similar technologies is likely to put a far greater strain on the nation's health care resources than is the practice of defensive medicine. These findings may be helpful to health policy makers, physicians, administrators, and legislators considering choices for health care reform in general and for medical liability reform in particular. PMID- 8723178 TI - Defensive medicine and tort reform: new evidence in an old bottle. AB - Quantitative analysis of medical liability's influence on medical practice is a small but growing field. The three foregoing articles illustrate three of the possible analytic approaches: case study of technological diffusion, survey of physician responses to detailed clinical scenarios, and multivariate analysis of the relation of physicians' scenario responses to objective liability experience. The articles also offer a good picture of the state of the art: Many difficulties hamper research in this area, and these articles, like others, offer considerable illumination but leave much uncovered. Defensive medicine surely exists, but its effects on health care spending and access are unclear. The most important lessons for public policy are that tort reform may be necessary but not sufficient to reduce the problems associated with defensive medicine, and that the major malpractice problem continues to be malpractice. PMID- 8723179 TI - Medical practice guidelines in malpractice litigation: an early retrospective. AB - The role of practice guidelines in malpractice litigation has been discussed in some theoretical detail. Little information is available, however, on how courts use guidelines or on the effort of state legislatures to explicitly link guideline compliance with malpractice defenses. We review all relevant case law and legislative enactments to shed light on the influence of medical malpractice on guidelines. We also use data from a nationwide survey of malpractice attorneys to supplement our legal analysis. Although guidelines are being used for both inculpatory and exculpatory purposes in common-law litigation (a two-way street), legislatures are interested in applying them only for exculpatory purposes (a one way street). PMID- 8723180 TI - Medical malpractice in Michigan: an economic analysis. AB - We analyzed the factors determining the amount of the recovery on claims based on medical malpractice. Our data set, which previously was not explored, consists of 20,428 claims arising within Michigan that were closed between 1978 and 1990. During this period, major changes were made in the law governing malpractice litigation. We determine the effect of these changes and of various other factors affecting medical malpractice claims. We analyze the effect of a statute that was designed to curtail "forum shopping" by attorneys for plaintiffs. This statute was initially successful in curbing such activity, but its effect seems to be diminishing. In addition, we compare mediation awards with settlement payments, and settlement payments with the expected value of claims in litigation. Evidence suggests that a mediation award is the mediation panel's estimate of a settlement payment appropriate for the case, and that cases are settled for substantially less than their expected value at trial. PMID- 8723181 TI - The Quinlan case revisited: a history of the cultural politics of medicine and the law. AB - This article explores the cultural politics of medicine and the law through a historical examination of the case of Karen Ann Quinlan. In viewing the Quinlan case as, in part, an unanticipated consequence of the 1968 redefinition of death, this article implicates the importance of historical perspective and methodology in examining the nuances of the cultural negotiation of professional power. Using popular, legal, medical, and bioethics sources in historical context, it reveals how the legal process can nurture misconceptions about medical practice and the role of technology. It also sheds light on how the search for legal protection can motivate medical behavior. In so doing, it challenges the view that Quinlan was an important gain for patients' rights. Instead it views Quinlan's chief legacy as its offer to the medical profession of freedom from criminal prosecution when removing life support from patients in a chronically vegetative state. PMID- 8723183 TI - ESOT update on immunosuppressive substances in clinical development or use 1995. AB - The ESOT Council conducted an inquiry on new immunosuppressive substances in order to help keep members of ESOT informed. Thirty-one pharmaceutical companies were sent a questionnaire indicating whether they were developing new immunosuppressive substances/antibodies. Sixteen companies responded: 11 furnished information on 16 substances; 5 said that they were not developing any new immunosuppressive agents. Fifteen companies did not reply at all. The results of the first inquiry are reported here. PMID- 8723184 TI - Magnetic resonance imaging for the evaluation of rejection of a kidney allograft in the rat. AB - Orthotopic DA (RT1a) into Lewis (RT1l) rat kidney allografts and control Lewis into-Lewis grafts were assessed by magnetic resonance imaging (MRI) and perfusion measurement after intravenous injection of a superparamagnetic contrast agent. MRI anatomical scores (range 1-6) and perfusion rates were compared with graft histology (rank of rejection score 1-6). Not only acute rejection, but also chronic events were monitored after acute rejection was prevented by daily cyclosporine (Sandimmune) treatment during the first 2 weeks after transplantation. In acute allograft rejection (n = 11), MRI scores reached the maximum value of 6 and perfusion rates were severely reduced within 5 days after transplantation; histology showed severe acute rejection (histologic score 5-6). In the chronic phase (100-130 days after transplantation), allografts (n = 5) manifested rejection (in histology cellular rejection and vessel changes), accompanied by MRI scores of around 2-3 and reduced perfusion rates. Both in the acute and chronic phases, the MRI anatomical score correlated significantly with the histological score (Spearman rank correlation coefficient rs 0.89, n = 30, P < 0.01), and perfusion rates correlated significantly with the MRI score or histological score (rs values between -0.60 and -0.87, n = 23, P < 0.01). It is concluded that MRI represents an interesting tool for assessing the anatomical and hemodynamical status of a kidney allograft in the acute and chronic phases after transplantation. PMID- 8723185 TI - Prolongation of lung xenograft survival in rats with a short course of deoxyspergualin and cyclosporin A. AB - The efficacy of 15-deoxyspergualin (DSG), cyclosporin A (CyA), and splenectomy- alone or in combination--in prolonging the survival of concordant lung xenotransplants was studied in the hamster-to-rat model. In the untreated group, rejection occurred within 3 days, with an elevation of lymphocytotoxic antibody titers. The rejected lung revealed that ED1 + cells were more prevalent than MRC OX8 + cells in the perivascular infiltrates. In the DSG group, the antibody response was suppressed and median survival increased to 7.5 days. The rejected lungs demonstrated a highly significant depression in ED1 + cellular infiltration and a moderate MRC OX8 + cellular infiltration. When maintenance CyA was combined with a short course of DSG, survival dramatically increased to beyond 100 days. There were no deposits of IgM, IgG, or C3 or of any cell infiltrate in the grafts of two animals sacrificed 107 and 119 days post-transplantation. We conclude that initial treatment with DSG combined with continuous CyA can suppress acute rejection in the hamster-to-rat lung xenograft model, resulting in long-term graft survival. PMID- 8723186 TI - Evaluation of a new solution containing trehalose for twenty-hour canine lung preservation. AB - We examined the efficacy of two new preservation solutions containing trehalose- an extracellular type (ET-K) of solution and an intracellular type (IT-K) of solution--in relation to that of Euro-Collins (EC) solution in 20-h canine lung preservation. Canine lungs were flushed with one of the three solutions (n = 5 for each solution) after pretreatment with PGE1 (20 micrograms/kg) and were stored for 20 h at 4 degrees C. The left lungs were transplanted and evaluated to 6 h post transplant. In the ET-K group, the arterial oxygen tension after reperfusion was significantly higher than in the IT-K and EC groups. The pulmonary vascular resistance, wet/dry weight ratio, and histological evaluation of each transplanted lung in the ET-K group were also better than in the IT-K and EC groups. This indicates that ET-K solution is useful for 20-h preservation of canine lung grafts. PMID- 8723187 TI - BQ-123, a specific endothelin (ETA) receptor antagonist, prevents ischemia reperfusion injury in kidney transplantation. AB - We studied the effects of the specific endothelin (ETA) receptor antagonist, BQ 123, on reperfusion injury in a rat model of kidney transplantation. First, Sprague-Dawley rats were divided into three groups: a sham nephrectomy (SNEPH), an autotransplantation (AUTO-Tx), and an allotransplantation (ALLO-Tx) group. In a fourth group, ALLO-Tx + BQ, allografts were flushed with 20 micrograms BQ-123 containing cold Ringer's lactate before transplantation. For the allograft groups, kidneys from white Wistar albino rats were transplanted into allogeneic Sprague Dawley recipients. Grafts were allowed 120 min of reperfusion after 40 min of cold ischemia. ET-1,2 plasma concentrations in the renal venous blood, and kidney tissue prostaglandin (PG) E2 and leukotriene (LT) B4 levels were studied. Diene conjugates (DC), hydroxyalkanals (HAA), hydroxyalkenals (HAE) and malondialdehyde (MDA) levels, as the products of lipid peroxidation, and protein carbonyls (PC) and protein sulphydryls (PS), as the parameters of protein oxidation, were also analyzed in the kidney tissue. Plasma ET concentrations increased significantly in the AUTO-Tx and ALLO-Tx groups (P < 0.05 and P < 0.01, respectively) but this increase was reversed in the ALLO-Tx + BQ group. None of the lipid peroxidation products except DCs (P < 0.05) increased in the AUTO-Tx group, whereas they all increased in the ALLO-Tx group (P < 0.01). Protein oxidation parameters also changed significantly (P < 0.01) in the ALLO-Tx group but did not in the AUTO-Tx group (P < 0.05). The differences in PGE2 and LTB4 levels were not significant. Histopathologic examination revealed prominent glomerular and tubular injury in the AUTO-Tx and ALLO-Tx groups but less in the ALLO-Tx + BQ group. In the last group, all parameters of lipid peroxidation (P < 0.001 for all) and PCs decreased, and PSs were preserved (P < 0.001 for both) when compared with the AUTO-Tx and ALLO-Tx groups. We conclude that BQ-123, in addition to inhibiting the binding of ET-1,2 to the ETA receptor, may also inhibit the release and/or synthesis of ET-1,2 and prevent reperfusion injury in kidney transplantation. PMID- 8723188 TI - Guidelines for donor selection and an overview of the donor operation in living related liver transplantation. AB - Guidelines for donor selection and an overview of the donor operation are reported on the basis of our experience with 120 cases of living related liver transplantation (LRLT) in pediatric patients. Once the parents had clearly expressed their desire to serve as donors, tests were performed to functionally and anatomically screen the donor livers to determine whether or not the parents' general physical condition allowed them to serve as donors. We then evaluated which of the two parental candidates was more suitable as a donor. The wishes of the family as to which parent should serve as donor was considered secondary and taken into account only in a few cases in which certain functional and/or anatomical abnormalities were uncovered that made the prime candidate less suitable. For the 120 LRLTs, 135 candidates were evaluated as potential donors, 15 (11.1%) of whom were rejected for various reasons. The mean volume of blood loss during the donor operation decreased significantly from 489 g in the first 60 LRLTs to 390 g in the latter 60 LRLTs; this was accompanied by a significant decrease in the mean volume of autologous blood transfused from 449 g to 390 g. Mean cold ischemia time of the graft increased significantly from 71.4 to 128.0 min, while mean operation time conversely decreased from 6.7 to 6.2 h. Bile leakage from the cut surface of the remnant liver, which was the only post operative surgical complication encountered, was noted in five cases. We conclude that donor candidates should be strictly selected according to basic guidelines, taking into account both the results of preoperative screening and the wishes of the family. With this accumuled experience, we have been able to simplify our LRLT operative procedure, resulting in decreases in blood loss volume, blood transfused, and operation time. PMID- 8723189 TI - Dopamine, dopexamine and dobutamine in liver transplant recipients: a comparison of their effects on hemodynamics, oxygen transport and hepatic venous oxygen saturation. AB - The purpose of this study was to determine the effects of vasoactive treatment with dopamine (DO), dopexamine (DX), and dobutamine (DOB) on hemodynamics, oxygen transport and hepatic venous oxygen saturation (SvhO2) after orthotopic liver transplantation (OLT). A pulmonary artery catheter was inserted into the right hepatic vein of 17 OLT patients. Timed infusion of DO, DX, and DOB was performed at the following rates: DO at 4 and 8 micrograms/kg per minute, DX at 4 and 8 micrograms/kg per minute, and DOB at 5 and 10 micrograms/kg per minute. Hemodynamics, oxygen transport variables, and SvhO2 were assessed. Each catecholamine induced a significant increase in cardiac index, oxygen delivery, and SvhO2. Mean arterial pressure was increased during DO and DOB, but significantly reduced during DX. Each inotrope increased oxygen delivery in parallel with SvhO2, suggesting a corresponding increase in hepatic oxygen supply. Therefore, it appears that each vasoactive drug may be utilized in OLT patients to provide oxygen delivery without impairment of splanchnic oxygenation. PMID- 8723190 TI - Factors influencing plasma lipid profiles including lipoprotein (a) concentrations in renal transplant recipients. AB - Fasting plasma cholesterol, triglycerides, high-density lipoprotein (HDL) and apoprotein (apo) B were elevated in 214 nondiabetic renal transplant recipients when compared to a reference group. Apo (a) was slightly but not significantly lower in transplant recipients (median 118 mg/dl, range 16-1680 vs 130 mg/dl, 10 1176) and this difference could be predicted from Lp (a) isoform analysis. Cholesterol, triglyceride, apo B and apo (a) concentrations correlated negatively with creatinine clearance but none of these parameters showed a significant association with proteinuria. Patients treated with steroids had higher plasma HDL concentrations than those receiving cyclosporin monotherapy (P < 0.01). The use of diuretics was associated with raised triglycerides (P < 0.001) and cholesterol (P < 0.01) and with reduced HDL (P < 0.01) whilst patients receiving beta-blockers had significantly higher triglycerides (P < 0.01) and lower HDL levels (P < 0.02). In multiple regression analysis, age (P < 0.01), creatinine clearance (P < 0.05) and diuretic therapy (P < 0.005) were independent risk factors for increased cholesterol whilst apo (a) levels correlated negatively with creatinine clearance (P < 0.005). These results suggest that impaired renal function, steroids and non-immunosuppressive drugs contribute to lipid abnormalites in renal transplant recipients. PMID- 8723191 TI - Extracorporeal membrane oxygenation for lung transplant recipients with primary severe donor lung dysfunction. AB - Primary severe donor lung dysfunction (DLD) is a significant complication after lung transplantation (LTx), and a high mortality is reported with conventional therapy. The purpose of this report is to review the experience of the University of Pittsburgh with extracorporeal membrane oxygenation (ECMO) for primary severe DLD after LTx. From September 1991 to May 1995, 220 LTx were performed at our center. Eight patients (8/220 = 3.6%) with severe DLD after LTx required ECMO support. The age of LTx recipients was 44 +/- 5 years (mean +/- SD); seven patients were female and one was male. Indications for LTx were: chronic obstructive pulmonary disease in four patients, bronchiectasis in two, and pulmonary hypertension in two. There were three single LTx and five bilateral LTx. The interval from LTx to institution of ECMO was 5.6 +/- 3.2 h (range 0-10 h). Three patients were supported with veno-venous (v-v) ECMO and five had veno arterial (v-a) ECMO. The duration of ECMO support was 7.3 +/- 4.8 days (range 3 15 days). activated glotting time (ACT) was maintained between 110 and 180 s with intermittent use of heparin. Seven patients (7/8 = 87%) were successfully weaned from ECMO and six patients (6/8 = 75%) were discharged home; they are currently alive after a follow-up of 17 +/- 10.1 months. One patient died on ECMO support for refractory DLD and another died 2 months after ECMO wean from multisystem organ failure. At 6 months follow-up, forced expiratory volume in 1 s (FEV1) is 2.35 +/- 0.91 (75% +/- 17.4% predicted) and mean forced vital capacity (FVC) is 2.53 +/- 0.81 (64% +/- 14% predicted). We conclude that ECMO can be lifesaving when instituted early after primary severe DLD. The v-v ECMO support is preferred when the patient is hemodynamically stable and adequate long-term function of the allograft is anticipated. PMID- 8723192 TI - High mortality from urothelial carcinoma despite regular tumor screening in patients with analgesic nephropathy after renal transplantation. AB - Patients with end-stage renal failure due to analgesic nephropathy have an increased risk of developing a urothelial carcinoma. To determine the impact of renal transplantation on the frequency of urothelial carcinomas, we analyzed 2072 patients who underwent 2371 renal transplantation between 1968 and 1993, including 78 (3.8%) with clinically proven analgesic nephropathy. Before and after transplantation a regular tumor screening was performed in patients with analgesic nephropathy by urine cytology and abdominal sonography. In 11 of the 78 patients with analgesic nephropathy (14.1%; age 51-66 years, 40-108 months after initiation of dialysis treatment, 5-77 months after transplantation), a urothelial carcinoma of the native urinary tract, especially the kidneys, was diagnosed. Therapy comprised nephroureterectomy (n = 6), transurethral resection (n = 6) and/or cystectomy (n = 2). Seven patients died due to tumor progression 16.3 (4-33) months postoperatively and one patient died due to a perioperative complication. Despite regular tumor screening after transplantation, the diagnosis of a urothelial carcinoma was made very late, leading to a high tumor related mortality. As a consequence, we suggest that a bilateral nephroureterectomy should be performed prophylactically in patients with proven analgesic nephropathy. In addition, a cystoscopy with lavage cytology testing of the bladder should be performed twice a year. PMID- 8723193 TI - Definitions of cytomegalovirus disease after heart transplantation: antigenemia as a marker for antiviral therapy. AB - In this prospective study, cytomegalovirus (CMV) antigenemia was defined as the marker for initiation and episodes of antigenemia as the indicator for the duration of antiviral therapy (CMV hyperimmune globulin and ganciclovir). The CMV antigenemia assay and CMV-specific IgM and IgG antibody tests were used to monitor CMV infection in 22 heart transplant recipients who, between October 1992 and July 1994, were followed up for 6 months. A total of 178 out of 627 antigenemia assays tested positive. The highest number of positive cells was greater after primary infection than after either reactivation (43.3 vs 0.3; P < 0.01) or reinfection (43.3 vs 9.3; P = NS). Sixty episodes of antigenemia were observed. More episodes of antigenemia were seen after primary infection than after either reactivation (4.6 vs 0.2; P < 0.01) or reinfection (4.6 vs 2.2; P = NS). The detection of antigenemia indicated the initiation of antiviral therapy within 24 h after the blood sample was harvested. Therapy was stopped immediately after a subsequent negative result became available. Our experience indicates that antigenemia directed antiviral therapy prevents CMV disease after primary and secondary infection in heart transplant recipients. PMID- 8723194 TI - How many endomyocardial biopsies are necessary in the first year after heart transplantation? AB - Since 1989, the immunosuppressive regimen used in all heart transplant (HTx) patients at our center has consisted of a combination of cyclosporin, azathioprine, and prednisone. No prophylactic cytolytic agents have been given. One hundred consecutive patients were followed for periods of 4-56 months (mean 27 months). The incidence of rejection was so low in the initial 18 patients that we felt confident about reducing the number of routine endomyocardial biopsies (EMBs) that were performed. The mean number of EMBs in this subgroup was 10 (median 11). In the next 20 patients, EMB was performed routinely on only three occasions during the 1st post-transplant year (at 2, 4, and 8 weeks). In the subsequent 62 patients, EMB was performed on post-transplant days 10, 20, 30, and 60. Further EMBs were performed after acute rejection episodes had been treated. No noninvasive methods of diagnosing rejection were employed. In 82 consecutive patients, therefore, the mean number of EMBs within the 1st year was five per patient (median four), with 58% undergoing fewer than five EMBs and 25% requiring more than five EMBs. In the entire group of 100 patients, the mean number of EMBs was 5.9. The incidence of acute rejection requiring increased therapy was 24%. Only 7% required i.v. steroids, two of whom (2%) also required ALG and/or OKT3, with 17% requiring increased oral immunosuppression alone. Actuarial survival was 98% at 30 days, 94% at 1 year, and 92% at 2 years. It is possible that we may have missed acute rejection episodes that resolved spontaneously. However, the excellent medium-term results would suggest that any such rejection episode did not progress to become hemodynamically significant. It may be, therefore, that when an effective immunosuppressive regimen is utilized, the number of EMBs performed at many centers is excessive. PMID- 8723195 TI - Cytomegalovirus infection and colonic perforation in renal transplant patients. AB - Cytomegalovirus (CMV) infection in immunocompromised patients is a major cause of morbidity and mortality. A well-documented manifestation of gastrointestinal CMV infection is gastrointestinal haemorrhage. In contrast, CMV-associated intestinal perforation has rarely been reported after transplantation, although it is well documented in AIDS patients. Three patients are reported who received their first cadaveric renal transplant in 1994 and subsequently developed CMV disease. During the course of their CMV illness, which was treated with ganciclovir, each presented with clinical suspicion of peritonitis and proceeded to laparotomy. All three were found to have sigmoid colon perforations with histological evidence of CMV infection. Following bowel resection and defunctioning, two patients made an uneventful recovery and have had the continuity of their bowel restored, but one died of overwhelming sepsis within hours of surgery. The explanation for the apparent clustering of this rare condition in transplant patients is uncertain. PMID- 8723196 TI - Living unrelated renal transplantation: the new alternative? PMID- 8723197 TI - JAK/STAT eats the fat. PMID- 8723198 TI - Transmitter timecourse in the synaptic cleft: its role in central synaptic function. AB - The speed of clearance of transmitter from the cleft influences many aspects of synaptic function, including the timecourse of the postsynaptic response and the peak postsynaptic receptor occupancy. The timecourse of transmitter clearance can be estimated either by detailed theoretical modelling, or from the attenuation of synaptic transmission produced by a low-affinity competitive antagonist. These approaches have been applied to several classes of central synapse, and results obtained are in broad agreement. The average concentration of transmitter peaks in the range 1-5 mM, and clearance is biphasic, with time constants of approximately 100 microseconds and 2 ms. The pulse of transmitter, although very brief, can prolong the timecourse of the fastest AMPA synaptic currents, and is sufficient to saturate postsynaptic GABA, glycine or NMDA receptors. PMID- 8723199 TI - Schizophrenia, the heteromodal association neocortex and development: potential for a neurogenetic approach. AB - The heteromodal association neocortex is believed to be a major site of involvement in schizophrenia. This system includes the prefrontal cortex and portions of the superior temporal and inferior parietal cortices, which are linked in cognitive networks observing complex executive functions. The heteromodal cortex is highly elaborated in humans and is believed to continue to develop past birth. The neuropathology of schizophrenia is likely to be heterogeneous and appears to involve developmental abnormalities, with a prominent genetic component. However, the genes involved in the development of the neocortex, and particularly the heteromodal cortex, are not well understood. A candidate-gene approach to schizophrenia using techniques of differential expression might now be feasible and could illuminate the basic neurobiology of the heteromodal cortical network. PMID- 8723200 TI - Gene-targeting studies of mammalian behavior: is it the mutation or the background genotype? AB - Gene targeting to create null mutations in mice is a powerful new tool in biology which will allow the molecular dissection of complex phenotypes such as mammalian brain function, and learning and memory. However, the attempt to interpret the phenotypical changes which arise in null-mutant mice is subject to several caveats. For example, the ability to disrupt a single gene in a targeted manner might lead one to overlook the effects of other genes. Ignoring the genetic background might lead to misinterpretation of results: the present article will demonstrate that the phenotypical abnormalities attributed to the null mutation in several molecular neurobiological studies could simply result from the effects of background genes. PMID- 8723201 TI - Unusual behavioral phenotypes of inbred mouse strains. PMID- 8723202 TI - Mice, gene targeting and behaviour: more than just genetic background. PMID- 8723203 TI - Gene-targeting studies: new methods, old problems. PMID- 8723204 TI - Neural simulation programs. PMID- 8723205 TI - Selective agonists for GABAC receptors. PMID- 8723206 TI - The TiPS/TINS Lecture. Catecholamines: from gene regulation to neuropsychiatric disorders. AB - In addition to their ability to change the electrical properties of neurons, evidence suggests that neurotransmitters are able to alter the cell's metabolism. Transmitter phenotype is labile and expression might be regulated, during development, by the cellular environment of neurons. The study of a key enzyme in the synthesis of catecholamines, tyrosine hydroxylase (TH), has provided clues about these adaptive responses. This enzyme has a large molecular diversity, resulting from the differential splicing of its mRNA, which is tissue-specific and might result in long-term changes in activity of the enzyme and, therefore, in the availability of neurotransmitter at various synapses. The presence of different DNA sequences at the TH locus confers susceptibility to various disorders of the brain, including manic-depressive illness and schizophrenia. Indeed, an association between a rare variant allele of the gene encoding TH and the occurrence of schizophrenia has been found in several populations. New techniques being developed to treat diseases such as Parkinson's disease involve various gene therapies, including a method of transferring genes directly into nerve cells using an adenovirus-based system. PMID- 8723207 TI - Interactions between self-motion and depth perception in the processing of optic flow. AB - Moving and acting in a 3D environment requires the perception of its 3D structure. Vision is known to play a crucial role in the control of self-motion, particularly through the changes in the retinal image subsequent to movements of the observer. Reciprocally, signals related to self-motion can also influence our visual perception of 3D space. These interactions between 3D visual perception and self-motion, as demonstrated behaviourally, are now better understood thanks to the development of computational models for processing moving images. They also bear a particular interest in the context of the recent intensive exploration of the inferior parietal lobe (IPL) by neurophysiologists. The IPL is now firmly established as one site of interaction between 3D visual perception and motor control. The parallel between behaviour and neurophysiology leads to a set of crucial, yet unanswered, questions. PMID- 8723208 TI - Neuronal networks for induced '40 Hz' rhythms. AB - A fast, coherent EEG rhythm, called a gamma or a '40 Hz' rhythm, has been implicated both in higher brain functions, such as the 'binding' of features that are detected by sensory cortices into perceived objects, and in lower level processes, such as the phase coding of neuronal activity. Computer simulations of several parts of the brain suggest that gamma rhythms can be generated by pools of excitatory neurones, networks of inhibitory neurones, or networks of both excitatory and inhibitory neurones. The strongest experimental evidence for rhythm generators has been shown for: (1) neocortical and thalamic neurones that are intrinsic '40 Hz' oscillators, although synchrony still requires network mechanisms; and (2) hippocampal and neocortical networks of mutually inhibitory interneurones that generate collective 40 Hz rhythms when excited tonically. PMID- 8723210 TI - Clinical practice guidelines. PMID- 8723211 TI - The challenge of health outcomes. AB - The health outcomes initiative can be seen as another passing phase in health care management or taken as a serious challenge to the planning, management and evaluation of health services. This paper explores those challenges. Implementation of the health outcomes initiative will require the application of valid, reliable and appropriately sensitive measures, the use of a broad approach to research, development and monitoring in such a way that it is an intrinsic part of service delivery, the adoption of policy and practice that is firmly based on evidence of outcomes, and the development of an approach to research that emphasises generalizability. PMID- 8723212 TI - RANZCP guidelines for psychotropic drugs: application to hospital inpatients. Royal Australian and New Zealand College of Psychiatrists. AB - The objective of this study was to monitor compliance with the Royal Australian and New Zealand College of Psychiatrists (RANZCP) guidelines for psychiatric drug use, in the forensic section of a psychiatric institution in Queensland. The author performed an audit of the charts and medical records of 65 patients in three forensic hospital wards at the John Oxley Memorial Hospital, a 73-bed forensic/secure section of the Wolston Park Hospital Complex. It was found that many patient records were lacking some relevant documentation. There was a higher rate of prescription for multiple psychotropic drugs than is recommended for patients at discharge from inpatient units. This quality assurance exercise has provided a basis for discussion to find ways to more closely comply with the RANZCP Guidelines for Psychotropic Drugs in Psychiatric Practice. No benchmarks are available for equivalent continuing inpatients in a similar setting. It has been shown that, for continuing inpatients in a forensic hospital, there is poor compliance with Draft Clinical Indicator No. 2.1 for Pilot Test (developed jointly by the RANZCP and the ACHS). Perhaps the threshold for this indicator needs to be raised in this clinical setting. The situation may be clarified when the proposed follow-up audit has taken place. There were no patients receiving doses in excess of currently-accepted limits, that is, Clinical Indicator (CI) No. 2.2 was 0, and there were 31 patients (47.7%) concurrently receiving two or more antipsychotic drugs and CI no.2.3 had a value of 31. PMID- 8723213 TI - Clinical indicators in colorectal surgery. AB - Two studies conducted in the state of Victoria have tested potential clinical indicators and the suggested thresholds for resection of colorectal carcinoma where an anastomosis has been performed. These studies involving 535 patients were independent of one another: one hospital based and one surgeon based. Threshold figures for these draft indicators have been compared with the study figures and found to be similar. It is suggested that wound infection (elective operation without formation of a stoma), anastomotic leak (clinically recognized) and mortality (elective operations in patients under the age of 80 years) are the most appropriate clinical indicators of colorectal resection for carcinoma. PMID- 8723214 TI - Mortality soon after discharge from a major teaching hospital: linking mortality and morbidity. AB - In this pilot study, data held by the Registrar of Births, Deaths and Marriages was linked to the Patient Master Index at St Vincent's Hospital Melbourne in order to identify early post discharge deaths, and inpatient records were linked with the Victorian Inpatient Minimum Database. Between July 1991 and December 1993 there were 831 deaths within 28 days of discharge. Injuries and poisoning were the commonest cause of death in younger patients who had attended the emergency department. These were responsible for 64% of deaths under the age of 40 years. There were deaths from cardiovascular disease which had been unrecognised during the hospital stay. There is also the need to identify in hospital those at risk of suicide. Amongst inpatients who were discharged home or transferred, the risk of death was greatest in the days immediately following discharge. Policies relating to the transfer of patients need to be reviewed as the study found that 30% of patients who had been transferred to long term care facilities died within four days. In conclusion, linking mortality and morbidity data can highlight areas of hospital care where there is the potential to prevent premature death, and improve management. The outcome of coronial investigations should be transmitted to hospitals for quality assurance purposes. PMID- 8723215 TI - Alfred Hospital Coronary Care Unit: an acute myocardial infarction quality assurance study. AB - A six-month prospective study was performed to assess the efficacy of delivering thrombolytic therapy to patients with acute myocardial infarction and admitted to the coronary care unit. Patient characteristics, time from chest pain onset to presentation to emergency department, from presentation to thrombolytic therapy, transfer from emergency department to coronary care unit major interventions and outcome were assessed. One hundred and twenty patients were admitted with acute myocardial infarction, mean age 66 years (26-91), 69% were males. Of these, 50% received thrombolytic therapy. The mean time from chest pain onset to emergency department was 192 +/- 164 minutes, transfer from emergency department to coronary care unit was 195 +/- 150 minutes. The mean time from presentation to emergency department to receiving thrombolytics was 63 +/- 12 minutes. Streptokinase was the choice of thrombolytic in 97%. Thrombolytic therapy was administered in emergency department in 80% of cases. Thirty-eight (63%) patients received thrombolytic therapy within 60 minutes of presentation. Compared to the non-thrombolytic group, the thrombolytic group were younger, 63 vs 69 years, P < 0.01, presented earlier to hospital (192 vs 394 minutes, P < 0.0005), were transferred to coronary care unit sooner (195 vs 472 minutes, P < 0.001), and had more coronary angiograms (29 vs 23, P < 0.02) and PTCA performed 10 vs 3, P < 0.04. There were no significant differences in length of coronary care unit stay, length of hospital stay, patients receiving CABG or death. PMID- 8723216 TI - International health technology assessment meetings. PMID- 8723217 TI - Agonist self-inhibitory binding site of the nicotinic acetylcholine receptor. AB - A major focus of current research on the nicotinic acetylcholine receptor (AChR) has been to understand the molecular mechanism of ion channel inhibition. In particular, we put special emphasis on the description of the localization of the agonist self-inhibitory binding site. Binding of agonist in the millimolar concentration range to this particular site produces inhibition of the ion flux activity previously elicited by the same agonist at micromolar concentrations. Due to the similitude in the pharmacological and electrophysiological behavior in inhibiting the ion channel of both high agonist concentrations and noncompetitive antagonists, we first describe the localization of noncompetitive inhibitor binding sites on the AChR. There is a great body of experimental evidence for the existence and location of luminal high-affinity noncompetitive inhibitor binding sites. In this regard, the most simple mechanism to describe the action of noncompetitive inhibitors which bind to luminal sites and, by its semblance, the agonist self-inhibition itself, is based on the assumption that these compounds enter the open channel, bind to different rings within the M2 transmembrane domain of the receptor, and block cation flux by occluding the receptor pore. However, the existence of high-affinity nonluminal noncompetitive inhibitor binding sites is not consistent with the open-channel-blocking mechanism. Instead, the presence of the quinacrine locus at the lipid-protein (alpha M1) interface approximately 7 A from the lipid-water interface and the ethidium domain located approximately 46 A from the membrane surface in the wall of the vestibule open the possibility for the regulation of cation permeation by an allosteric process. Additionally, the observed (at least partially) overlapping between the quinacrine and the agonist self-inhibitory binding site also suggests an allosteric process for agonist self-inhibition. For this alternative mechanism, cholinergic agonist molecules first need to be partitioned into (or to be adsorbed onto) the lipid membrane to further interact with its binding site located at the lipid-protein interface. PMID- 8723218 TI - Functional reconstitution of KCl-evoked, Ca(2+)-dependent acetylcholine release system in Xenopus oocytes microinjected with presynaptic plasma membranes and synaptic vesicles. AB - We have developed a new method for the generation of functionally active presynaptic chimeras in Xenopus laevis oocytes. Frog oocytes injected with presynaptic subcellular fractions extracted from the electric organ of Torpedo marmorata release acetylcholine in a calcium-dependent manner upon chemical stimulation. Neither oocytes injected without presynaptic plasma membranes nor oocytes injected with ghost erythrocyte plasma membrane instead of presynaptic plasma membrane release acetylcholine. This suggests that specific presynaptic components necessary for KCl-evoked, Ca(2+)-dependent acetylcholine release become functionally integrated in the Xenopus laevis oocytes. Moreover, rhodaminated presynaptic plasma membranes and the synaptic vesicle protein synaptophysin are detected on the oocyte surface by fluorescence or immunofluorescence, respectively, showing that the injected presynaptic components are incorporated into the membrane of the frog oocyte. Furthermore, Botulinum neurotoxin type A, a specific blocker of acetylcholine release in the neuromuscular junction, inhibits the neurotransmitter release from the chimerical oocytes. This suggests that targets for toxin action are also functionally incorporated in the oocyte upon injection of membranous presynaptic components. Our results show that oocytes injected with presynaptic components behave as cholinergic nerve ending chimeras, at least in terms of neurotransmitter release and toxin targets. The system bypasses some problems associated with messenger RNA expression because not only proteins, but native presynaptic components are incorporated. This new technique may provide a useful approach for electrophysiological and pharmacological studies in order to characterize the synaptic transmission. PMID- 8723219 TI - Three different states of the chromatin structure of the mouse peripherin gene. AB - The chromatin structure of the mouse peripherin gene domain was analyzed in peripherin-positive and -negative cell lines. At least nine DNase I hypersensitive sites (HSS) are present within the 20-kb peripherin domain in the mouse neuroblastoma cell lines which express peripherin. Three of them are situated in intron I and intron III, the others being distributed within the 5' flanking region up to -5.5 kb. The presence of these sites was also investigated in the peripherin chromatin domain of peripherin-negative cell lines. Two other types of HSS distribution were observed along the peripherin gene according to the category of cell considered: constantly peripherin-negative cells, or negative cells arising from transiently peripherin-expressing precursors. From comparison of HSS patterns in these cell lines with those of neuroblastoma cells, it can be predicted that HSS located in the region -1500/+800 bp participate in cell-specific expression of the mouse peripherin gene. PMID- 8723220 TI - Induction of connexin43 and gap junctional communication in PC12 cells overexpressing the carboxy terminal region of amyloid precursor protein. AB - Previous studies have shown that PC12 cells overexpressing beta/A4 amyloid peptide display altered morphology characterized by pronounced membrane ruffling and extensive intercellular appositions. Having observed other cell types in which these features accompany increased connexin43 (Cx43) production and gap junctional communication, we examined Cx43 in normal and beta/A4-transfected PC12 cells. Studies of two beta/A4-transfected PC12 clones revealed an induction of Cx43 expression by Western blotting, intracellular and plasma membrane-associated Cx43 in some cells of cultures processed by immunofluorescence, dye-transfer between some cells microinjected with Lucifer Yellow, and gap junctions between cells examined by EM. Normal and vector-transfected PC12 cells exhibited none of these properties. Increased immunofluorescence in some clusters of beta/A4 transfected cells was also observed with a monoclonal antibody against connexin32. The results suggest that beta/A4 amyloid peptide may cause aberrant intercellular communication and gap junction formation through induction or increased expression of connexins in cells that are not normally coupled or only poorly coupled by gap junctions. PMID- 8723221 TI - Combined effects of GDNF, BDNF, and CNTF on motoneuron differentiation in vitro. AB - We have previously shown that glial cell line-derived neurotrophic factor (GDNF), in addition to promoting the survival of dopaminergic neurons in cultures from embryonic rat ventral mesencephalon,also increases the activity of choline acetyltransferase (ChAT) in the cranial motoneurons present in these cultures (Zurn et al.: Neuroreport 6:113-118, 1994). By using the intermediate filament protein peripherin as a motoneuron marker, we report here that GDNF increases the number of motoneurons as well as the length of their neurites. Brain-derived neurotrophic factor (BDNF) and ciliary neurotrophic factor (CNTF) also promote ChAT activity, motoneuron survival, and neurite outgrowth in these cultures, but to varying degrees. Although these three molecules have similar effects on cultured motoneurons, we provide evidence for a distinct mode of action of GDNF, BDNF, and CNTF, since combinations of GDNF and BDNF, GDNF and CNTF, and BDNF and CNTF have either additive or synergistic effects on ChAT activity and motoneuron number. In addition to the previously described motoneuron-specific neurotrophic factors BDNF and CNTF, GDNF combined with the latter two factors may provide an important tool for the treatment of human motoneuron diseases such as amyotrophic lateral sclerosis and spinal muscular atrophy, both by increasing efficiency of treatment, and by decreasing the likelihood of deleterious side-effects. PMID- 8723222 TI - Upregulation of B50/GAP-43 protein mRNA in rat dorsal root ganglia during cisplatin intoxication. AB - Expression of the growth-associated protein B50 (GAP-43) mRNA in dorsal root ganglia (DRG) of rats was studied by in situ hybridization. In response to treatment with the neurotoxic agent cisplatin, B50 mRNA expression was significantly enhanced following a cumulative cisplatin dose of 14 mg/kg. In the untreated age-matched control animals, only half of the ganglion cells exhibited expression of B50 mRNA (mean hybridization signal, 10 times background), whereas at a cumulative cisplatin dose of 14 mg cisplatin every neuron exhibited well above background expression (mean hybridization signal, 34 times background). Cotreatment with a neuroprotective ACTH4-9 analog known to prevent cisplatin neuropathy in rats did not affect the overall expression of B50 mRNA. However, in the subpopulation of large sensory neurons, B50 mRNA content was significantly higher in the group cotreated with the ACTH4-9 analog as compared with the saline cotreated group after 14 mg/kg of cisplatin. We conclude that in analogy with the well-known upregulation of B50 mRNA following mechanical nerve lesions, treatment with the neurotoxic drug cisplatin also leads to an increase in B50 mRNA expression. This observation lends strength to the hypothesis that in neuropathies an imbalance between regenerative and degenerative mechanisms exists. The ability of the larger sensory neurons to retain an increased B50 mRNA expression better after cotreatment with the peptide than without may be related to stimulation of regenerative processes by this ACTH4-9 analog. PMID- 8723223 TI - Pharmacological characterization of inositol-1,4,5,-trisphosphate binding to membranes from retina and retinal cultures. AB - Light and excitatory amino acids (EAA) stimulate the phosphoinositide cycle in the vertebrate retina. The regulation of Ca2+ release from intracellular stores by inositol-1,4, 5-trisphosphate (IP3) involves an interaction of this compound with specific receptors. By means of [3H]IP3-specific binding, we studied the kinetic and pharmacological properties of IP3 receptors in the chick retina as well as in primary cultures of neurons and glia from this tissue. The equilibrium time for the binding reaction was 15 min and was optimal at alkaline pH (8.3). IP3 receptor displayed high affinity (K(B) approximately 40 nM) and selectivity for D-IP3, compared to D-IP4 > L-IP3 > D-IP2 > D-IP1. These characteristics were the same in subcellular fractions from outer (P1) and thinner (P2) plexiform layers, binding sites being more abundant in P2 (2.65 pmol/mg protein). IP3 receptors were present in both neuronal and glial cultures, but were concentrated in neuronal cultures. Binding was not affected by ryanodine, or caffeine, related to calcium-induced calcium release (CICR) channels, nor by the endoplasmic reticulum Ca2+ ATPase inhibitor thapsigargin, while heparin affectively inhibited IP3 binding. GSSG and thimerosal increased the affinity of [3H]IP3 binding from IC50 approximately 80 nM to IC50 approximately 40 nM; this effect was reversed by DTT. Binding in zero Ca2+ was decreased by low concentrations of Ca2+ (350 nM). These results suggest that actions of IP3 in the retina are regulated by physiological changes in intracellular pH and Ca2+ concentrations, as well as by the oxidation state of the receptor. Additionally, the presence of IP3 receptors in Muller glia opens the possibility of IP3 participation in nonsynaptic signalling through Ca2+ waves in glial cells. PMID- 8723224 TI - Increased expression of brain-derived neurotrophic factor but not neurotrophin-3 mRNA in rat brain after cortical impact injury. AB - Levels of brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NT3) mRNA expression were measured in a rodent model of traumatic brain injury (TBI) following unilateral injury to the cerebral cortex. To obtain reliable data on the co-expression of neurotrophin genes, adjacent coronal sections from the same rat brains were hybridized in situ with BDNF and NT3 cRNA probes. BDNF mRNA increased at 1,3, and 5 hr after unilateral cortical injury in the cortex ipsilateral to the injury site and bilaterally in the dorsal hippocampus. NT3 mRNA did not change significantly following injury. Our results suggest that TBI produces rapid increases in BDNF mRNA expression in rat brain without changes in NT3 mRNA expression, a finding which differs from studies of ischemia and seizures. It is possible that increased levels of BDNF mRNA rather than NT3 are important components of pathophysiological responses to TBI. PMID- 8723225 TI - Major histocompatibility complex class I expression in oligodendrocytes induces hypomyelination in transgenic mice. AB - Increased expression of MHC Class I occurs in the central nervous system in association with demyelinating diseases such as multiple sclerosis and experimental allergic encephalomyelitis. To determine if MHC Class I expression by oligodendrocytes induces white matter pathology, the MHC Class I gene was expressed in transgenic mice under the control of the myelin basic protein (MBP) promoter. These mice display a neurological phenotype at 21 days-of-age. We examined these mice at 1,3, and 12 weeks-of-age. MHC Class I was detected in the brains and spinal cords of transgenic mice but not in control mice. Class I was located in oligodendrocyte perikarya but not in myelin sheaths. The central nervous system of these transgenic mice was hypomyelinated and contained hypertrophic microglia and astrocytes. These observations establish that Class I expression by oligodendrocytes delays normal myelination but does not cause inflammatory demyelination. This hypomyelinating animal model is of potential use in studying the interactions between immunologically active molecules and remyelination in disorders of myelin. PMID- 8723226 TI - Mitochondrial NADH dehydrogenase and CYP2D6 genotypes in Lewy-body parkinsonism. AB - The cause of nerve-cell death in sporadic Parkinson's disease remains unknown. Although environmental factors have been traditionally implicated in the etiology of Parkinson's disease, recent studies strongly suggest that there is a genetic contribution to this multifactorial disorder. We studied archival brain tissue from clinically and neuropathologically verified cases of Parkinson's disease, using nonradioactive cycle sequencing and restriction enzymatic analysis of polymerase chain reaction products. Twenty-one Parkinsonian brains with brain stem Lewy-bodies and 77 control brains were genotyped at two mitochondrial loci previously implicated in the etiology of neurodegenerative disease. In addition, genotyping was performed for two alleles of the debrisoquine 4-hydroxylase gene (CYP2D6). A heteroplasmic mtDNAG5460A missense mutation in the ND2 subunit gene of NADH dehydrogenase was three times more frequent in Parkinson cases (4/21) compared to controls (5/77). A homoplasmic mtDNAA4336G transition which alters the mitochondrial tRNAGln gene product was found in one Parkinson case. Frequencies of the CYP2D6G1934A and CYP2D6C2938T alleles were not significantly different between Parkinson cases and controls. Two Parkinsonian brains with high degrees of heteroplasmy for the ND2G5460A mutation and one CYP2D6C2938T homozygous case showed very high numbers of Lewy-bodies in the substantia nigra. The results of this study are in line with the concept that different genetic loci may be involved in Parkinson's disease susceptibility. They provide a hint that the ND2(5460) mutation, in combination with other factors, could play a role in disease pathogenesis in a subset of patients. PMID- 8723227 TI - Synaptic potentials recorded from sea anemone muscle cells in situ. AB - Three pulse types, A,B, and C, can be recorded from column ring preparations of the sea anemone Calliactis parasitica (Couch). Lucifer Yellow injection confirmed that the recording sites are intracellular in endodermal myoepithelial cells. Type C pulses are spontaneously active depolarizing pulses, arising from a resting potential of around -60mV, and are usually < 5 mV in amplitude. When the cells were voltage-clamped these pulses reversed at around 0 mV membrane potential; therefore, we conclude that they are synaptic potentials. In some cells an increase in frequency of type C pulses accompanied contraction of the circular muscle field. Inward currents in these myoepithelial cells are carried by Ca2+, and although Na(+)-free solutions did not affect inward currents they did eliminate type C pulses, i.e., these synaptic potentials may result from activity of Na(+)-dependent nerve cells. PMID- 8723228 TI - Characterization of Na+/H+ exchange activity in cultured rat hippocampal astrocytes. AB - Astrocytes actively maintain their intracellular pH (pHi) more alkaline than expected by passive distribution of H+. Acid extruding transporters such as the amiloride-sensitive Na+/H+ exchanger (NHE) are necessary for pH regulation. Currently, four mammalian NHEs (NHE1-NHE4) have been cloned, with a fifth (NHE5) partially cloned. We attempted to determine which isoform(s) of NHE was present in cultured hippocampal astrocytes using amiloride sensitivity and immunospecificity as criteria. In the absence of HCO3-, amiloride blocked pHi recovery after an acid load with an IC50 of approximately 3.18 microM, similar to values reported for the amiloride-sensitive isoforms NHE1 and NHE2. Immunoblotting with a highly specific antibody for NHE1 identified a 100 kDa protein, indicating the presence of NHE1 in whole brain, hippocampus, and cultured hippocampal astrocytes. Further probing for an additional amiloride sensitive NHE failed to detect evidence of the presence of NHE4. Surprisingly, application of the potent analog of amiloride, ethylisopropylamiloride (EIPA), caused a reversible alkalinization of pHi, suggesting the presence of an additional acid/base transport mechanism that is EIPA-sensitive. PMID- 8723229 TI - Strategies for using nonhuman primates in genetic research on multifactorial diseases. AB - Multifactorial diseases such as coronary heart disease, cancer, and osteoporosis are of increasing public health concern. Such diseases have a complex etiopathogenesis involving genetic and environmental factors and interactions between these factors. Nonhuman primates are uniquely suited as animal models for complex human diseases because of their close evolutionary relationship to humans. We present the rationale for using nonhuman primates as animal models in research designed to assess the genetic determinants of multifactorial diseases. Strategies and current approaches for the use of nonhuman primate models in genetic research on complex diseases are reviewed. PMID- 8723230 TI - Mechanisms regulating the reproduction and fertility of some mammalian species in their natural environments. PMID- 8723232 TI - Hepatic pathology resulting from mouse hepatitis virus S infection in severe combined immunodeficiency mice. AB - Mouse hepatitis virus (MHV) is a pervasive pathogen that causes morbidity and mortality in mouse colonies worldwide. Although it is not a major cause of mortality in immunocompetent mice, infections from MHV strains of lower virulence can be fatal to athymic nude mice. The histopathologic features and alterations of serum biochemical parameters resulting from infection with a low-virulence MHV strain in severe combined immunodeficiency (scid) mice has not been well described. Thus we recently studied the disease caused by MHV-S in scid mice after intranasal inoculation. Mouse hepatitis virus infection in scid mice, which have severe defects of B and T cells, may be highly lethal, resulting in immediate mortality. However, our results indicate that scid mice survived for an average of 12 to 14 days after infection with doses of MHV up to 10(7) PFU/mouse. The virus caused a significant increase in serum enzyme activities and bilirubin concentration associated with histologically demonstrable hepatocellular injury at postinoculation days 3, 4, and 8. Furthermore, virus was detected in mouse liver homogenates and nasal and bronchial lavage specimens. These results provide valuable information regarding the histopathologic and biochemical consequences of MHV-S infection in scid mice. PMID- 8723231 TI - Attachment glycoproteins and receptor specificity of rat coronaviruses. AB - Murine coronavirus (MHV) and rat coronavirus (RCV) are antigenically related viruses that have different natural rodent hosts. Both MHV and RCV can be propagated in the L2(Percy) and CMT-93 mouse cell lines. In these cell lines MHV uses the MHV receptor (MHVR or Bgp1a) and several related murine Bgp glycoproteins in the immunoglobulin superfamily as receptors. To determine whether RCV also uses these murine glycoproteins as receptors, we characterized the envelope glycoproteins of two strains of RCV and compared the effects of anti MHVR monoclonal antibody on susceptibility of the mouse cells to MHV and RCV. The Parker (RCV-P) and sialodacryoadenitis (RCV-SDAV) strains of RCV expressed the spike glycoprotein S, but only RCV-P expressed a hemagglutinin-esterase glycoprotein that had acetylesterase activity. Therefore RCV-SDAV must bind to cellular receptors by the viral S glycoprotein, whereas RCV-P might bind to cells by its hemagglutinin-esterase glycoprotein as well as by S. Pretreatment of L2(Percy) 41.a or CMT-93 cells with anti-MHVR monoclonal antibody blocked infection with MHV-A59 but did not prevent infection of these murine cells with RCV-P or RCV-SDAV. Baby hamster kidney cells transfected with cDNAs encoding MHVR (Bgp1a) or Bgp2 were susceptible to MHV-A59 but not to RCV-P or RCV-SDAV. Thus the RCV strains cannot use these murine coronavirus receptors and must be infecting murine cells by another, as yet unknown, receptor. PMID- 8723233 TI - Evaluation of cecal ligation as a model of mucoid enteropathy in specific pathogen-free rabbits. AB - Mucoid enteropathy is a serious disease of rabbits, the cause of which is unknown. Ligation of the cecum has been reported to cause a mucoid enteropathy like syndrome in 70% of conventional rabbits. The goal of this study was to evaluate this model of mucoid enteropathy in Pasteurella-free, coccidia-free rabbits for use in future studies. Five rabbits served as unoperated controls (group 1). Eight rabbits underwent ligation of the cecum, with large vessels and nerves spared (group 2). In six rabbits the distal branch of the ileocecocolic vessels and nerve were incorporated into the cecal ligature (group 3). At necropsy 3 to 5 days after surgery, all group-3 rabbits had copious amounts of clear, gelatinous mucus in the colon. Only one group-2 rabbit had grossly evident mucus hypersecretion, whereas none of the group-1 rabbits did. Group-3 rabbits had areas of necrosis in the cecum; this was not seen in group-1 or group-2 rabbits. Rabbits of groups 2 and 3 had inflammation of the distal portion of the colon. In specific-pathogen-free rabbits cecal ligation alone did not reliably stimulate mucus hypersecretion but induced a disease similar to natural mucoid enteropathy. Cecal ligation including vessels provided a reproducible syndrome of mucus hypersecretion; however, the severe cecal necrosis was not consistent with the naturally acquired disease. PMID- 8723235 TI - Use of polymerase chain reaction for diagnosis of type D simian retrovirus infection in macaque blood. AB - We developed a simple and specific polymerase chain reaction (PCR) method for the detection of type D simian retrovirus (SRV) infection (SRV-1, SRV-2, and SRV-3) using whole blood samples from macaques. Each pair of primers for the three serotypes of SRV was highly specific for its respective envelope proviral DNA and was sensitive enough to easily detect about five copies of the SRV-2 proviral genome. The PCR products were confirmed by Southern blot hybridization with digoxigenin-labeled internal oligonucleotide probes. For diagnostic purposes the three sets of primers were mixed together. The molecular weight of the PCR product for each of the three serotypes differed. Serotypes were confirmed by hybridization with a mixture of SRV-2 and SRV-1 and -3 internal probes. The PCR analysis of 39 whole blood samples correctly identified five SRV-1 and nine SRV-2 culture-positive samples. It also detected SRV-2 in two culture-negative blood samples from monkeys from which SRV had been previously isolated. PMID- 8723234 TI - Mucus secretagogue activity in cecal contents of rabbits with mucoid enteropathy. AB - Cecal filtrates from rabbits with cecal ligation-induced mucoid enteropathy have been reported to cause goblet cell hyperplasia in intestinal explants. This study was performed to see whether such filtrates would stimulate mucus secretion from intestinal explants. Filtrates were prepared from cecal contents of five control rabbits, five rabbits that had undergone cecal ligation, and five rabbits that had had the distal branch of the ileocecocolic vessels and nerve incorporated into the cecal ligature. After incubation of each with ileal and colonic explants from five healthy rabbits, the amount of mucus in the media/filtrate solution was measured with an enzyme-linked lectin assay. Significantly (P < 0.05) more mucus was secreted by colonic explants in the presence of cecal filtrates from either of the cecal ligation groups compared with controls. With pooled filtrates from each group, these results were confirmed by measuring secretion of radiolabeled mucus from explants that had been preincubated with [3H]glucosamine. The secretagogue activity was found to be precipitable by 85% ammonium sulfate and destroyed by heat treatment (100 degrees C for 30 min) or acid treatment (pH 1.0 for 30 min). Very little, if any, secretagogue activity was present in cecal vein serum. The results support the hypothesis that the cecal contents of rabbits with cecal ligation-induced mucoid enteropathy stimulate mucus secretion from colonic explants taken from healthy rabbits. PMID- 8723236 TI - Evaluation of various oral antimicrobial formulations for eradication of Helicobacter hepaticus. AB - Helicobacter hepaticus colonizes the cecum and colon of several strains of mice from a variety of commercial suppliers, persistently infects mice, causes chronic hepatitis, is linked to hepatic tumors in A/JCr mice, and has been associated with inflammatory bowel disease of athymic and scid mice. For this reason, eradication of the organism from infected mouse colonies is desirable. We recently reported that amoxicillin or tetracycline-based triple therapy (amoxicillin or tetracycline in combination with metronidazole and bismuth) given by oral gavage 3 times daily for 2 weeks eradicated H. hepaticus in 8- to 10-week old A/JCr mice. To establish a more convenient therapy regimen for eradicating H. hepaticus, we evaluated water and dietary administration of various antibiotic combinations in A/JCr and DBA/2 mice naturally infected with H. hepaticus. The A/JCr male mice received amoxicillin-based triple therapy in drinking water or by oral gavage, or received tetracycline-based triple therapy in the drinking water. The DBA/2J female mice received amoxicillin-based triple therapy in a specially formulated dietary wafer or by oral gavage, or received enrofloxacin in drinking water. All treatments were given for a 2-week period. Control animals received no treatment. One month after treatment, H. hepaticus was recovered from the liver, cecum, or colon of A/JCr control mice and mice receiving amoxicillin- or tetracycline-based triple therapy in drinking water but not in mice receiving amoxicillin-based triple therapy by oral gavage. Helicobacter hepaticus was not recovered from DBA/2J mice receiving amoxicillin-based triple therapy in dietary wafer or by oral gavage but was recovered from control mice and 7 of 10 mice receiving enrofloxacin in drinking water. Results indicate that amoxicillin-based triple therapy administered in the diet or by oral gavage is effective in eradicating H. hepaticus. Antibiotics administered in the water, however, were not effective in eradicating the organism. PMID- 8723237 TI - Immunoassay for detection of antibodies to simian immunodeficiency virus and human immunodeficiency virus in serum. AB - We developed a simple, inexpensive, rapid assay for the detection of antibodies to simian immunodeficiency virus (SIV) and human immunodeficiency virus type 1 (HIV-1) in serum. The immunoassay uses inactivated SIV and HIV-1 gp41 transmembrane recombinant protein as antigenic adsorbents on a nitrocellulose filter membrane. Diluted serum, with the addition of Protein-A-Gold, is gravity filtered through the filter membrane, blocked, and buffer-washed. Antibodies to HIV or SIV or both in serum bind to the appropriate antigen, and the resulting antigen-antibody complex reacts with Protein-A-Gold to produce a readable pink color. Field evaluation of the test on 30 human and 70 nonhuman primate sera in Kenya and Zaire indicated that the test had at least 93 and 90% correlation with Western blot sensitivity and specificity respectively. Prior refrigeration of the test kit and incubation of sera during testing were not required. This result indicates that the test may be a rapid, economical, and simple test for detecting HIV, SIV, or both in serum. This immunoassay can be useful for carrying out HIV and SIV serosurveys in countries with limited or no laboratory facilities. PMID- 8723238 TI - Measurement of insulin pulsatility by sampling directly from the portal vein: a surgical model for placement of long-term prehepatic vascular sampling catheters. AB - A practical and repeatable method for measurement of pulsatile insulin release in the dog was developed by direct, chronic cannulation of the portal vein cephalad to the pancrease and the entry point of the cranial pancreaticoduodenal vein, with subsequent exteriorization of the sampling catheter. Mixed breed male dogs of various body weights and ages underwent midline laparotomy for exposure of the portal vein system. After exposure the portal vein was dissected free and cannulated through a purse-string suture placed just caudad (approximately 1 cm) to the entry point of the cranial pancreaticoduodenal vein with a cuffed, medical grade, silastic catheter. The catheter was advanced cranial to a point just distal to the level of the liver and secured in place via the purse-string suture and a single stay suture. The catheter was then passed through the dorsolateral abdominal wall and routed subcutaneously to a point between the shoulder blades. After catheter plug attachment the catheter was tested for patency, flushed, heparin locked, then was secured to the subcutaneous tissue between the scapulas. The dorsal skin wound and abdominal incision were closed in a routine manner, and the dogs recovered from anesthesia. Catheter patency and function in all dogs were maintained for periods up to 4 weeks. No postoperative complications such as catheter tract infections occurred, maintenance was minimal, and all dogs were afforded complete freedom of movement and activity throughout the study, with no requirement for cumbersome jackets or wraps. PMID- 8723239 TI - Vascular anatomy of experimentally induced left varicocele in the rat. AB - A variety of techniques have been used by various investigators to study the effects of experimentally induced left varicocele in the rat model. It has become apparent that information about the vascular anatomy of this condition in the rat is deficient, though such information could be important to understanding the model. We have examined the left and right venous architecture serving the testes in rats with and without experimentally induced left varicocele. The left spermatic vein has four consistent collaterals that are tributary to the left iliac vein. The right spermatic vein does not have these consistent collaterals, but many times a collateral develops from the right spermatic vein to either the right iliac vein or the distal part of the caudal vena cava. When this occurs, the remaining length of the right spermatic vein becomes a minor effluent vessel. Eleven irregularly observed vessels were identified, none of which existed on the left and only two of which were observed on the right. Experimentally induced left varicocele is associated with dilatation of the left spermatic vein and all collaterals to the left iliac vein. The condition does not cause dilatation of the right venous system, but one collateral was identified that communicated between the left and right sides via the vesicular plexus. It is speculated that the differences between left and right sides in the number of consistent collaterals might be due to the patterns of embryologic development, which differ somewhat between the left and right spermatic veins. PMID- 8723240 TI - P-wave duration in a clinically normal hound population. AB - Electrocardiogram (ECG) analysis is a common noninvasive technique used for diagnosis of cardiac disease in the clinical and research branches of veterinary medicine. Accurate analysis of P-wave duration, amplitude, and morphology is crucial to identification of morphologic and functional changes of the atria. The published accepted maximal normal value for P-wave duration in the dog is < or = 40 milliseconds. We looked at P-wave duration in ECG obtained as part of routine quarantine health screening over a period of 1 year in 364 clinically normal hounds weighing 13 to 35 kg. The dogs were neither anesthetized nor sedated and were placed in standard position. P-wave duration was classically determined from the lead-II recording. Mean P-wave duration for all dogs (44.9 +/- 6.1 milliseconds) was greater than published accepted normal values for the dog. There was a significant difference in mean P-wave duration by body weight (P < 0.001); dogs weighing > or = 20 kg had longer mean P-wave durations than dogs weighing < 20 kg (45.3 and 41.6 milliseconds respectively). There were also significant differences in mean P-wave duration by sex (P < 0.01), with a greater mean duration for females (45.4 milliseconds) than for males (43.8 milliseconds). All other ECG parameters were within published accepted normal values. A P-wave of prolonged duration leads to a diagnosis of abnormalities in cardiac morphology and/or function. Published accepted normal values for P-wave duration, at least for a clinically normal hound population, appear to be shorter than the true normal values. An error in published accepted normal standards may lead to overdiagnosis of cardiac abnormalities, as well as to erroneous results in cardiovascular studies. Therefore we recommend that the standard for P-wave duration be increased above the currently accepted standard of < or = 40 milliseconds. PMID- 8723241 TI - Cardiopulmonary and analgesic effects of xylazine, detomidine, medetomidine, and the antagonist atipamezole in isoflurane-anesthetized swine. AB - The cardiovascular and respiratory effects of three alpha(2)-adrenergic agonists (xylazine 2mg/kg of body weight; detomidine, 40 micrograms/kg; medetomidine, 40 micrograms/kg) and their specific antagonist, atipamezole (200 micrograms/kg) were examined in young, isoflurane-anesthetized (1.3% end-tidal concentration) swine (weight range, 15 to 35 kg). The intravenous administration of all three alpha(2)-agonists caused an initial significant (P < 0.05) but short-lived increase in arterial blood pressure. Atipamezole also increased blood pressure, and this effect persisted throughout the period of observation. All agonists caused a sustained significant bradycardia, whereas atipamezole significantly increased heart rate (30 +/- 7 beats per min). The cardiac index tended to transiently decrease 5 to 10 min after agonist injection (significant only for xylazine at 2 min after injection) from an average pre-injection value of 166 ml/kg per min and did not change in response to atipamezole. None of the drugs significantly modified arterial blood gas (PaO2, PaCO2) or pH values. Xylazine and medetomidine but not detomidine or atipamezole manifested short-lived analgesic properties in response to clamping of the interdigital fold. PMID- 8723242 TI - Use of the novel carbocyanine tracer fast-DiI for investigating upper respiratory tract cranial nerves in prenatal rats. AB - The use of traditional neuroanatomic tracing methods with tracers such as horseradish peroxidase requires living systems to take up and transport the label. These tracers have limited application in the study of prenatal systems. The advent of the carbocyanine dye DiI provided a tool by which neuronal pathways may be traced in postmortem fixed tissue. This dye allows careful dissection of prenatal organisms and specific application of the tracer to the neuroanatomic structure under investigation. Although DiI has become increasingly popular, it is limited by the difficulty in applying crystals to peripheral nerves and in the lengthy trace times, particularly in fixed tissue. A chemically modified version of DiI has been introduced that may overcome some of the limitations in using this tracer. The newer dye, fast-DiI, is easy to apply and can trace neural pathways in a shorter period. We describe our experience with the use of fast-DiI in the prenatal rat for the investigation of motoneurons that subserve upper respiratory tract structures. We have determined protocols for fixation, application of dye, processing of tissue, and visualization of traces. The entire protocol can be completed within 1 week, and the use of fast-DiI is easy to learn and apply. The resultant labeling of traced nerves is specific and clearly demonstrates respective motor nuclei and individual motoneurons. PMID- 8723243 TI - Diagnostic exercise: peracute death in a research dog. PMID- 8723244 TI - Diagnostic exercise: granulomatous encephalitis in guinea pigs. PMID- 8723245 TI - Noninvasive versus invasive blood pressure measurement in normotensive and hypotensive baboons. PMID- 8723246 TI - Measurement of milk yield and constituents in mice by a unilateral teat occlusion method. PMID- 8723247 TI - Use of cryopreservation techniques of embryos and spermatozoa for production of transgenic (Tg) mice and for maintenance of Tg mouse lines. PMID- 8723248 TI - Superovulation, fertilization, and in vitro embryo development in BALB/cByJ, BALB/cJ, B6D2F1/J, and CFW mouse strains. PMID- 8723249 TI - Quantitative evaluation of a technique for harvesting peritoneal macrophages from normal and laparotomized rats. PMID- 8723250 TI - Tail sectioning: a rapid and simple method for repeated blood sampling of the rat for corticosterone determination. PMID- 8723251 TI - Isolations of Potosi virus from mosquitoes collected in the United States, 1989 94. AB - Potosi (POT) virus, a recently characterized Bunyamwera serogroup virus, was discovered when it was isolated from Aedes albopictus collected at a waste-tire site in Potosi, Washington County, Missouri, during 1989. During the following year, POT virus was not isolated from 39,048 mosquitoes, including 17,519 Ae. albopictus, collected in Washington County. In 1991, mosquito collections from South Carolina, Ohio, and Michigan yielded 8 strains of POT virus: 6 from Coquillettidia perturbans and one each from Culex restuans and Psorophora columbiae. Additional collections of Ae. albopictus from several states during 1990-93 failed to yield further isolates of POT virus. In 1994, POT virus was isolated from Ae. albopictus and Anopheles punctipennis in North Carolina and from Ae. albopictus in Illinois. These represent the first virus isolations of any type for Ae. albopictus in those states. Thus far, POT virus has been isolated from 5 mosquito species in different genera in 6 states. The known geographic range of POT virus, based on virus isolations, has been extended from Missouri to the upper Midwest and the Atlantic seaboard. Potential vector relationships and possible transmission cycles of POT virus are discussed. PMID- 8723252 TI - Elevational distribution of mosquitoes in a mountainous area of southeastern Wyoming. AB - During the summer of 1984, mosquito sampling with CDC miniature light traps and standard mosquito dippers was used to investigate the elevational distribution of different species of mosquitoes. Of 27 species found in significant numbers in the study area in southeastern Wyoming, 8 were found primarily in the lower elevations (2,134 and 2,591 m). Nine additional species were found in both the middle and lower elevations (2,134-3,048 m), whereas 8 occurred only in the middle areas (2,592-3,048 m). A single species (Aedes punctor) was found in both the middle and upper elevations (2,439-3,292 m), and yet another species (Aedes impiger) was found primarily in the upper (alpine) area, from 3,049, to 3,292 m. There is some evidence to indicate that restriction in elevational distribution is a result of habitat specificity. PMID- 8723253 TI - Evaluation of cattle insecticide treatments on attraction, mortality, and fecundity of mosquitoes. AB - Attraction, engorgement, mortality, and fecundity were observed on host-seeking Aedes vexans and Psorophora confinnis given the opportunity to feed on insecticide-treated steers in stable traps. Individual steers were treated with Ectrin ear tags, Ectrin spray, Terminator ear tags, Saber ear tags, or Ivomec injection or were left untreated. Aedes vexans and Ps. confinnis accounted for over 98% of 20,738 mosquitoes collected. Relative attraction ranged from 0.45 with Saber ear tag to 0.84 with Terminator ear tag. Engorgement rates of mosquitoes from insecticide-treated steers ranged from 55 to 91%. Engorgement was reduced 18-45% by Terminator ear tag, Ectrin spray, and Saber ear tag treatments. Forty-eight percent mortality of unengorged Ae. vexans and 61% mortality of unengorged Ps. confinnis was observed at collection with the Saber ear tag treatment. Percentage of mortality of engorged mosquitoes of both species was generally much lower. Ivomec injection treatment showed the greatest effect on delayed (48-h) mortality of engorged mosquitoes, with 41 and 82% mortality for Ae. vexans and Ps. confinnis, respectively. No insecticide treatment had a significant effect on fecundity. PMID- 8723255 TI - Effect of tadpole shrimp, Triops longicaudatus, (Notostraca: Triopsidae), on the efficacy of the microbial control agent Bacillus thuringiensis var. israelensis in experimental microcosms. AB - Laboratory bioassays using Culex quinquefasciatus larvae evaluated the effect of tadpole shrimp on the persistence of Bacillus thuringiensis var. israelensis (B.t.i.) in water collected from the surface of field microcosms. Time elapsed since B.t.i. treatment, as well as presence or absence of soil and tadpole shrimp, affected B.t.i. persistence at the water surface in 15- and 30-cm total depths. The presence of tadpole shrimp slowed the natural decline in B.t.i. effectiveness over time, but this effect was depressed when soil was present. Tadpole shrimp foraged throughout the water column and stirred up the substrate, keeping more particles in suspension at the surface than in microcosms with no shrimp, in microcosms with water depths of 15 and 30 cm. PMID- 8723254 TI - Oviposition preferences of Culex pipiens and Culex restuans for infusion-baited traps. AB - The number of egg rafts oviposited by Culex pipiens and Culex restuans in infusion-baited ovitraps varied with the type of infusion substrate, infusion age, method of infusion preparation, and calendar date. In one or more tests, more Culex egg rafts were collected from water infused with Kentucky bluegrass sod, mixed grass clippings, straw, or a commercial rabbit chow than from a water control. Infusions prepared with oak leaves, maple leaves, and sod soil were ineffective as Culex oviposition attractants; however, alfalfa infusions ranged from marginally effective to unattractive. Rabbit chow infusions required one or more reinfusions before becoming active oviposition lures when prepared in unconditioned (unused) ovitraps. Infusions prepared in conditioned ovitraps (those that had previously contained the same type of infusion) had shorter delay periods. Effective oviposition lures for Culex species included continuous infusions, such as sod and sod grass, and those in which the substrate was infused in the water for only a brief period (24-48 h), such as infusions of rabbit chow and mixed grass clippings. The percentage of egg rafts from Cx. restuans was usually greater in sod and grass infusions than in rabbit chow infusions, whereas Cx. pipiens showed a slight preference for rabbit chow infusions over sod and grass infusions. The temporal shift in relative oviposition activity of the 2 Culex species (cross-over) varied from year to year and between sites in one year. PMID- 8723256 TI - Evaluation of controlled-release mosquito repellent formulations. AB - Eight polymer and 9 microcapsule formulations of deet were tested on laboratory rabbits against Aedes aegypti and Anopheles albimanus. Several formulations were significantly more effective than simple (unformulated) deet at the same strength for periods up to 24 h. Best results were obtained with a polymer formulation containing a high molecular weight fatty acid and 3 microcapsule formulations containing lanolin, gum arabic, gelatin, tannic acid, stearic acid, polypropylene glycol, water, and a commercial lotion in the microcapsule and carrier fractions. PMID- 8723257 TI - Deposition of malathion and permethrin on sod grass after single, ultra-low volume applications in a suburban neighborhood in Michigan. AB - Deposition of malathion and permethrin onto grass surfaces, after ultra-low volume (ULV) application, was studied in a suburban neighborhood in Saginaw County, Michigan. Commercial concentrates of malathion (Cythion ULV) and permethrin (Biomist 4 + 12) were sprayed using a truck-mounted ULV aerosol generator. Sod-grass blocks (0.18 m2) were placed in the frontyard and backyard of homes in the neighborhood at 4 distances to 91.4 m from the road where applications were made. Grass samples were taken from the sod blocks before application and at 15 min, 12 h, 24 h, and 36 h after application. Samples were extracted with solvent, and extractions were subjected to gas-liquid chromatography for detection of malathion and permethrin. Ranges of detection for malathion were 0.0-16.6 mg/0.18 m2 and for permethrin were 0.0-25.9 mg/0.18 m2. Most detections were from samples taken nearest the road at 15 min after application. Detections declined as a logarithmic function of time after application and as an exponential function of distance from the road. PMID- 8723259 TI - Human host avidity in Aedes albopictus: influence of mosquito body size, age, parity, and time of day. AB - Diel patterns of human host attack by Aedes albopictus in the laboratory were studied in relation to mosquito body size, age, parity, and time of day. Analysis of responses in 10-, 15-, and 20-day-old females indicated a significant main effect due to time of observation in the diel period, as well as significant time x parity and time x age interactions. The distribution of mean host attack responses during the diel period was bimodal with approximately 70% of all activity during photophase (0800-2000 h); attack rates were highest in the morning (0800 h) and evening (1400-2000 h) and lowest between 0200 and 0600 h. The diel pattern of attack responses was bimodal for nulliparous and parous females, but parous females were more active than nulliparous females between 1400 and 2000 h. This pattern became increasingly bimodal during photophase, as mosquitoes aged, regardless of mosquito body size or parity. Variations in host avidity patterns between young and old females suggest that mosquito repellent bioassays initiated early in the day, that last > or = 6 h, or that use young females (approximately 5 days old) overestimate the protection period of deet against mosquitoes > 10 days old. PMID- 8723258 TI - Observations on nocturnal biting activity and host preference of anophelines collected in southern Thailand. AB - Over a 13-month period, 5,127 adult female mosquitoes were collected on human bait during multiple collecting periods between 1800 and 0600 h in 5 villages in southern Thailand. There were marked differences in the biting activity of different species at each of the villages studied. Anopheles maculatus and Anopheles sawadwongporni were collected most often at dusk or in the first hours of darkness between 1800 and 2100 h. Although specimens of Anopheles dirus were collected consistently between 1900 and 0400 h, peak collections were made between 2000 and 2300 h. Anopheles minimus was collected consistently throughout the night without a clearly discernible peak. Of more than 2,400 Anopheles mosquitoes collected in CDC light traps, 133 specimens (5.4%) contained blood, nearly all of which was identified by enzyme-linked immunosorbent assay (ELISA) to be of bovine origin. Ten specimens contained blood from more than one host species. PMID- 8723260 TI - Population size, parity structure, and wing length of Coquillettidia perturbans in an Ohio focus of eastern equine encephalitis. AB - Adult female density, parity status, and wing length were determined weekly for a population of Coquillettidia perturbans in an area enzootic for eastern equine encephalitis virus in central Ohio. Samples were collected in CO2-baited CDC miniature light traps from the first week in June through the 2nd week of September 1992. Population density indicated a single emergence peak during the 2nd week in July. However, parity rates showed 2 peaks, occurring in the first week of August (70.9% parous) and the 2nd week of September (55.3% parous), which suggested that there was a relatively small 2nd generation. Average wing length declined significantly over the season. The decline in size was negatively correlated with average air temperature occurring at least 6 wk before the time of emergence. Despite the seasonal decline in wing length, the low coefficient of variation for the average wing length (5.5) indicated relatively little variation in size. Comparison of parous and nulliparous female wing lengths each week suggested that there was no association between size and survival in this species. PMID- 8723261 TI - Essential oil analysis and field evaluation of the citrosa plant "Pelargonium citrosum" as a repellent against populations of Aedes mosquitoes. AB - A plant recently introduced into North America as the citrosa, Pelargonium citrosum ('Van Leenii'), has been marketed as a biological repellent against mosquitoes. Citrosa is claimed to repel mosquitoes within a 10 ft.2 (0.93 m2) area due to a continuous fragrant release of citronella oil. The total essential oil yield was 0.2 +/- 0.1% from fresh plant material. Chemical analysis by the authors revealed that combined essential oils of fresh greenhouse- and field grown citrosa have 35.4 +/- 6.2% geraniol, 10.4 +/- 1.6% citronellol, 8.9 +/- 2.0% isomenthone, and 6.8 +/- 3.8% linalool. Both the morphology and essential oil of citrosa fall within the Pelargonium x asperum hybrid complex and are similar to 'Rose', the commercial rose geranium. No character of morphology or essential oil of a Cymbopogon species yielding commercial citronella oil could be detected in the citrosa. The effectiveness of the citrosa as a repellent against field populations of spring Aedes spp. mosquitoes was evaluated and compared with a 75% deet (N,N-diethyl-3-methylbenzamide) formulation. Deet provided > 90% reduction in mosquitoes biting subjects for up to 8 h post-treatment. There was no significant difference between citrosa-treated and nontreated subjects. PMID- 8723262 TI - Seasonal abundance and parity rates of Anopheles species in southern Thailand. AB - Mosquitoes were collected with human bait, animal bait, and CO2-baited light traps during a 13-month period in 5 villages located along the Myanmar border in southern Thailand. Altogether, 11,608 adult females consisting of 21 anopheline species were collected. Abundance of species varied according to the village where collections were made. In one village that was studied most intensely, Anopheles minimus was the most commonly collected species, representing 55.6, 10.6, and 56.5% of the human bait, cattle bait, and light trap collections, respectively. In addition, An. minimum was abundant throughout the year, whereas Anopheles maculatus was most commonly collected in July and August coincident with the rainy season. Anopheles sawadwongporni, abundant at the beginning and the end of the rainy season, was most commonly collected in June. Anopheles dirus was most abundant in April; however, it represented only 3.6% of all mosquitoes collected that month. Of 813 females collected in Palao-U Village and dissected, 38.9% were parous. PMID- 8723263 TI - Contact toxicity of permethrin-impregnated military uniforms to Culex pipiens (Diptera:Culicidae) and Phlebotomus papatasi (Diptera: Psychodidae): effects of laundering and time of exposure. AB - The effects of laundering and time of exposure on the insecticidal activity of military uniform fabric impregnated with permethrin at 0.125 mg active ingredient (AI)/cm2 are reported from susceptibility tests with laboratory-reared mosquitoes, Culex pipiens, and sand flies, Phlebotomus papatasi. Knockdown/mortality resulting from exposure of groups of female insects for periods of 1, 3, 5, 7, and 10 min was recorded and compared among 5 different treatment/wash groups (untreated/unwashed, treated/unwashed, treated/1-wash, treated/2-wash, treated/3-wash). Laundering was by machine washing with detergent and warm water followed by hot-air machine drying. Post-exposure assessments at intervals from 0 to 60 min and at 24 h showed that knockdown for each exposure time and wash group was initially low but increased steadily during the first hour post-exposure. Sand flies were less sensitive than mosquitoes to knockdown during the first 60 min after contact with treated/unwashed fabric; however, 24-h mortality rates for sand flies were higher as compared with mosquitoes. The permethrin remaining after a 3rd wash had little knockdown effect on mosquitoes but was toxic to sand flies at each of the 5 exposure times. Significant reductions in the knockdown effectiveness of permethrin-treated fabric to mosquitoes and sand flies was associated with single and repeated washings of the fabric. PMID- 8723264 TI - Evaluation of light traps combined with carbon dioxide and 1-octen-3-ol to collect anophelines in Venezuela. AB - A 6-month study was carried out in northcentral Venezuela to evaluate the efficiency of the CDC light trap and the updraft ultraviolet (UV) light trap combined with carbon dioxide (CO2) or 1-octen-3-ol (or both) and human baits to sample outdoor Anopheles aquasalis and Anopheles albimanus populations. The human baits caught far more mosquitoes than did any of the other trapping methods. Comparing each of the trapping methods to the human bait catches, UV light trap + CO2 gave a closer correspondence of the ratio of An. aquasalis to An. albimanus compared with the ratio found in human baits than did any of the other trapping methods. The mean parous rate was significantly lower in human bait catches than in all of the trapping methods except for An. aquasalis in UV light trap with CO2. We consider the UV light trap with CO2 to be the most reliable substitute for human bait catches. PMID- 8723265 TI - Metaphase karyotypes of Anopheles of Thailand and Southeast Asia: V. The Myzomyia Series, subgenus Cellia (Diptera:Culicidae). AB - Metaphase karyotypes of 9 species of the Myzomyia Series show intra- and interspecific differences based on quantitative variation and distribution of constitutive heterochromatin in the sex chromosomes or the centromeric regions of the autosome pairs or both. Anopheles jeyporiensis and Anopheles aconitus each exhibit 4 forms of mitotic karyotypes, which may reflect interspecific differences within each taxon. The well-defined genetic species within the Anopheles minimus and the Anopheles culcifacies complexes clearly exhibit distinctive metaphase karyotypes that can be used as diagnostic characters for separating these sibling species, which are difficult to identify by morphological criteria alone. Our analysis on metaphase karyotypes of Anopheles pampanai, Anopheles varuna, and Anopheles flavirostris also confirms their morphological identification based on heterochromatin differences in the sex chromosomes and autosome 2. PMID- 8723266 TI - Long-term (1980-94) population trends of pestiferous Chironomidae (Diptera) along a lakefront in central Florida. AB - Populations of adult chironomids occurring along 5-6 km of waterfront in the city of Sanford facing Lake Monroe, central Florida, were monitored from January 1980 to December 1994 using New Jersey light traps. The annual mean number of total midges per trap per day ranged from 269 (1994) to 8,009 (1980). Among the more than 20 species of midges occurring in the traps, Glyptotendipes paripes was the most abundant, followed by Chironomus crassicaudatus. These 2 species comprised 95.6% of total midges collected. Annual cycles of midge abundance were positively correlated with air temperature. Maxima of most species occurred in late spring/early summer; G. paripes peaked in late summer. Year-to-year midge population levels showed significant inverse correlations with Lake Monroe water depth and annual rainfall in the Sanford area. PMID- 8723267 TI - Organophosphate resistance in Culex pipiens from Cyprus. AB - Populations of Culex pipiens were sampled from 8 locations in Cyprus between 1987 and 1993. All population samples generally revealed organophosphate resistance to malathion, temephos, chlorpyrifos, fenthion, dichlorvos, and pirimiphos methyl, in decreasing order of magnitude. Of 7 populations assessed with the carbamate propoxur, all proved to be resistant to different degrees. Of the 6 populations tested with permethrin, 2 were resistant to permethrin. Resistance was associated with the presence of 5 different overproduced esterases (esterases A1, A2, A5, B2, and B5) as well as an insensitive form of acetylcholinesterase. These results are discussed in relation to the ongoing mosquito abatement program in Cyprus and to similar programs in other parts of the world. PMID- 8723268 TI - Attraction of Aedes albopictus adults in sod infusion. AB - Aedes albopictus adults were recovered from sod-baited gravid traps at 7 sites in 5 counties of southern and southwestern Illinois in areas ranging from suburban to rural. The attraction of this species to a lure usually associated with members of the Culex pipiens complex was investigated under laboratory and field conditions. Olfactometer and oviposition preference tests in the laboratory showed that Ae. albopictus is attracted to sod infusion and females readily oviposit on substrates in contact with the infusion. Both males and females responded to the sod infusion in the olfactometer. The attraction of multiple mosquito species to sod-baited gravid traps was demonstrated at a waste tire site in Jasper County, Illinois. Females of Ae. albopictus, Aedes triseriatus, and Culex species were collected from gravid traps placed along the edge of woods at distances ranging from 100 to 200 m from the tire site. Only a small percentage of the Ae. albopictus females appeared to be gravid (less than 5%), unlike the majority of Culex and Ae. tristeriatus females. PMID- 8723269 TI - Rapid decomposition of permethrin in the outer fly of an experimental tent in Pakistan. AB - The residual activity of permethrin on the canvas of an experimental tent, used by nomadic refugees in a malarious area in northern Pakistan, was assessed. A permethrin emulsion sprayed on the inner sheet of a pitched double-fly tent (0.5 g/m2) had an effective residue for at least 6 months in bioassays using the local malaria vectors, Anopheles stephensi and Anopheles culcifacies. However, a high concentration of permethrin (2 g/m2) sprayed on the inner surface of the outer tent, simulating single-fly tents, decomposed within 2 months. These findings were confirmed with chromatographic analysis. Under the conditions found in the study area, the shortened residual effect most likely resulted from photodecomposition, not previously reported to affect operational performance of this insecticide. PMID- 8723270 TI - Notes on the mosquitoes of Nepal. IV. Results of the 1994 collecting in the Midwestern Region, including new country records and voucher confirmation (Diptera, Culicidae). AB - During 1994, field studies were conducted in the Midwestern Region of Nepal. Two camps were located in the "inner terai," low mountain valleys between the Churia Range and the Mahabarat Lekh. A third camp was in the mountains at Jumla. Visits were made to 2 high mountain sites, Simikot, Humla District, and Rara National Park. The result from all these sampling sites was the recovery of 6 new country records, 5 in the genus Aedes and 1 in the genus Heizmannia. Some mosquitoes in the Nepal checklist had no voucher specimens. Locality data are given for 9 of these. Biodata on another species that is quite rare in Nepal are given. PMID- 8723272 TI - Fecundity, fertility, and developmental stage survivorship of Eretmapodites quinquevittatus. AB - This laboratory study analyzed fecundity, fertility (percentage hatched), and developmental stage survivorship of Eretmapodites quinquevittatus. Fecundity for females averaged 190.4 eggs, whereas paired (one male/female pair) females averaged 100.5 eggs. Fertility for autogenous eggs from paired females was 84.4%, which was significantly greater than percentage hatched of anautogenous eggs (67.3%). Percentage hatched of autogenous eggs from experimental groups (25 males/25 females) was 87.8%, whereas percentage hatched of anautogenous eggs was 74.5%. Developmental stage survivorship was significantly different for every stage of development of progeny from autogenous eggs, except the egg stage, when comparing experimental groups with pairs. Developmental stage survivorship of progeny from anautogenous eggs was significantly different only at the egg stage and 1st instar when comparing experimental groups and pairs. PMID- 8723271 TI - Detection of multiple bloodfeeding in Culiseta melanura using a histologic technique. AB - We used a histologic technique to detect multiple bloodfeeding in a single gonotrophic cycle by Culiseta melanura. In a laboratory study with colonized mosquitoes, 77% (10/13) of known multiple meals were detected when the interval between meals was 24-30 h. Outside that range, known multiple meals were undetectable with this method. No multiple blood meals were detected in 653 wild engorged mosquitoes collected during 3 consecutive years from the Pocomoke Swamp in Maryland. Although previous studies have shown that Cs. melanura will feed twice in the same evening, it rarely, if ever, takes multiple blood meals at 24-h intervals. Our study also indicates that a thorough laboratory standardization is required prior to application of the histologic technique to species for which it has not been studied. This would include a time-series analysis to define species specific limits for detecting known multiple meals. PMID- 8723273 TI - Evaluation of the laboratory rabbit model for screening topical mosquito repellents. AB - The laboratory rabbit was evaluated as a model for screening topical mosquito repellents, using data obtained in tests of deet (N,N-diethyl-3-methylbenzamide) against Aedes aegypti on humans and rabbits. Host-specific differences in the action of the test material were quantified by multiple regression analysis. The test material was less effective but more persistent in tests on rabbits, and responses of the mosquito test population were more variable. PMID- 8723274 TI - Efficacy and persistence of Altosid pellets against Culex species in catch basins in Michigan. AB - Larvae of Culex pipiens and Cx. restuans in catch basins were exposed to Altosid pellets (4% active ingredient, [S]-methoprene) applied at a rate of 11.3 kg/ha (7 g of pellets per catch basin). Under field conditions, the pellets yielded an average 82% emergence inhibition of adult mosquitoes over the 15-wk trial period. PMID- 8723275 TI - Anopheles fluviatilis complex: host feeding patterns of species S, T, and U. AB - The host feeding specificity of Anopheles fluviatilis sibling species S, T, and U was determined by analyzing blood meal source using countercurrent immunoelectrophoresis. A distinct difference in the feeding patterns was observed among these species. Species S was found to be predominantly anthropophagic with 91% of the population having fed on human blood. In contrast, species T and U were either exclusively or primarily zoophagic. PMID- 8723276 TI - Volunteers needed for study of HIV viral load test. PMID- 8723277 TI - The "epidemic" of breast cancer in the U.S.--determining the factors. AB - Breast cancer incidence rates in the United States rose by 24% between 1973 and 1991. Mortality during this period, however, remained stable. Both the 5-year relative survival rate and the rates of in situ and stage I breast cancers have been increasing, while the incidence of later-stage cancers has been decreasing. Increased mammography screening may explain the documented jump in breast cancer incidence rates during the mid-1980s. Differences in the distribution of breast cancer risk factors may account, in part, for the temporal trends in breast cancer incidence. In particular, breast cancer risk factors may vary by birth cohort, including age at menarche, age at first birth, physical activity, obesity, diet, alcohol intake, estrogen therapy, and exposure to environmental organochlorines. After decades of epidemiologic research, a preventive approach to breast cancer that focuses on the physiologic effects of the sex steroid hormones, and their potential interactions with family history, is being carefully formulated. PMID- 8723278 TI - Palliative pelvic exenteration: patient selection and results. AB - Total pelvic exenteration is a radical abdominoperineal operation designed to treat locally extensive pelvic malignancy. In the past, the morbidity and mortality has been such that this procedure was considered justified only in the curative setting. As experience has increased and operative survival has improved, the indications for radical pelvic surgery have expanded to include palliation of the symptoms of pelvic tumors, including pain, obstruction, fistulas, and bleeding. The careful selection of suitable candidates for palliative pelvic exenteration requires an extensive preoperative evaluation of the patient's underlying physical and emotional health, as well as an assessment of the local and distant extent of the tumor. In appropriately selected patients, significant palliation of the symptoms associated with an uncontrolled pelvic malignancy can be accomplished with substantial improvement in quality of life. PMID- 8723279 TI - Clinical trials referral resource. Cancer chemoprevention and control trials. PMID- 8723280 TI - Do families understand "do not resuscitate" orders? PMID- 8723281 TI - Post-transplantation complications of unrelated bone marrow. PMID- 8723282 TI - Fox Chase/Stanford study: radiation therapy effective in treating younger men with early-stage prostate cancer. PMID- 8723283 TI - Pathologic evaluation of prostatic carcinoma: critical information for the oncologist. AB - During the last few years, pathologists have assumed several important roles in the assessment of adenocarcinomas of the prostate. The establishment of postoperative serum prostate-specific antigen (PSA) levels as a sensitive indicator of progression following radical prostatectomy has provided the impetus to correlate pathologic findings with progression following surgery. The advent of the thin needle biopsy gun and screening with serum PSA has dramatically increased the number of needle biopsies performed to rule out prostatic adenocarcinoma. With these very limited tissue samples, pathologists are challenged to make diagnoses, as well as to accurately grade and quantify the tumor. This article explores topical issues in the pathology of prostate adenocarcinoma, with particular emphasis on practical aspects of pathologic evaluation that oncologists need to know for patient management. PMID- 8723284 TI - Regular NSAID use appears to reduce breast cancer severity. PMID- 8723285 TI - Monoclonal antibodies may help locate and treat occult metastases. PMID- 8723286 TI - New sequential dosing regimen shows promise in node-positive breast cancer. PMID- 8723287 TI - MoAb A33 shows promise in targeting colon cancer for radioimmunotherapy. PMID- 8723288 TI - Intensified VICE regimen may improve SCLC survival. PMID- 8723289 TI - Nonionizing electromagnetic fields and cancer: a review. AB - Low-frequency electromagnetic radiation had previously been thought to cause human injury only by generation of excess heat or by shock from direct contact with electric current. Information accumulating over the past few decades, however, suggests that nonionizing electric and magnetic fields associated with this radiation may be an environmental etiology in human disease. Human beings are affected not only by natural background nonionizing electromagnetic fields produced by the Earth, but also by a host of manmade sources. Of the diseases believed related to these fields, cancer and participation of these fields in the carcinogenic process have received considerable attention. This paper is a review of the basic science that points to this possible association. PMID- 8723290 TI - Transition to managed care will bring changes for oncologists, need for guidelines, says ACCC panel. PMID- 8723291 TI - Oncology carve-outs another managed care option. PMID- 8723292 TI - MoAb to block growth signals plus chemo may act synergistically to kill tumor cells. PMID- 8723293 TI - Drug-resistant pneumococci increasingly common in elderly, study suggests. PMID- 8723294 TI - ABMT recommended for relapsed Hodgkin's disease, but better overall survival not yet proved. PMID- 8723295 TI - Studies to determine if IL-2 can prevent relapses in post-transplant NHL and AML. PMID- 8723296 TI - The outpatient management of febrile neutropenia in cancer patients. AB - Treatment of fever and neutropenia in cancer patients has been recognized for 30 years as a medical emergency, requiring prompt in-hospital evaluation and institution of broad-spectrum intravenous (i.v.) antibiotics. This action was deemed necessary due to the high frequency of life-threatening infections in febrile neutropenic patients, with no way to distinguish patients who are infected from those who are not. In recent years, it has become clear that not all neutropenic cancer patients are at the same level of risk for developing severe infections or life-threatening complications during neutropenia. Those who are at low risk may be candidates for treatment outside the hospital setting, either with i.v. regimens or potent oral antibiotics. The identification of low risk febrile neutropenic patients and the specific outpatient approaches that have been tested to date are discussed. Outpatient management of fever during neutropenia could obviously be much less costly than standard inpatient care and could improve quality of life for low-risk patients undergoing cancer therapy. PMID- 8723297 TI - Antibody with two targets may boost immunotherapy benefit. PMID- 8723298 TI - NSAIDs may prevent colon cancer through apoptosis, not anti-inflammatory effects. PMID- 8723299 TI - Time-dependent effects of a haloperidol challenge on energy metabolism in the normal human brain. AB - Positron emission tomography and the fluorodeoxyglucose method were used to measure regional brain metabolism before and 2 h after haloperidol (5 mg, i.m.) in 11 young normal men. These data were compared with measures obtained from nine previously studied normal men who had received no drug intervention. Although a previously published study had demonstrated significantly decreased metabolism in whole brain, neocortex, limbic cortex, thalamus, and caudate nucleus 12 h after a 5-mg dose of haloperidol, the present 2-h study did not show significant metabolic changes despite the fact that significant extrapyramidal effects occurred. Taken together, these studies demonstrate differences in the temporal organization of behavioral and metabolic responses to haloperidol challenge. PMID- 8723301 TI - Hypometabolism in olfactory cortical projection areas of male patients with schizophrenia: an initial positron emission tomography study. AB - Several studies have reported olfactory deficits in schizophrenic patients. This study examines local cerebral metabolic rate within two cortical areas in eight normal men and eight schizophrenic men. A significantly greater degree of hypometabolism was observed in the schizophrenic men in the cortical area of the nondominant hemisphere that receives direct uncrossed olfactory projections. PMID- 8723300 TI - A controlled positron emission tomography study of obsessive and neutral auditory stimulation in obsessive-compulsive disorder with checking rituals. AB - Ten nondepressed patients with obsessive-compulsive disorder (OCD) who were characterized by predominant checking rituals were compared with 10 age- and sex matched control subjects. Hemispheric and regional cerebral blood flow levels (rCBF) were measured with positron emission tomography (H2 15O) across four conditions: rest, auditory stimulation with idiosyncratic normal or abnormal obsession, auditory stimulation with neutral verbal stimuli, and rest. Order of neutral and obsessive stimulation was randomized. Higher subjective responses to obsessive than to neutral stimulation were found in both groups; subjective response was higher in OCD patients when obsessive stimulation was presented first. A four-way analysis of variance (group x stimulation order x hemisphere x condition [neutral or obsessive stimulation]) was performed on stimulation minus rest normalized rCBF values. Control subjects had significantly higher rCBF in the thalamus and putamen. A trend toward higher rCBF in OCD patients was found in the superior temporal regions. When neutral stimulation was presented first, rCBF was significantly higher in the caudate region of control subjects. Obsessive stimulation was associated with higher rCBF than neutral stimulation in orbitofrontal regions in both groups of subjects. Under obsessive stimulation, superior temporal and orbitofrontal activities were correlated in OCD patients but not in control subjects. Our study suggests specific abnormalities of information processing in the basal ganglia and temporal structures of compulsive checkers. PMID- 8723302 TI - MRI changes in schizophrenia in late life: a preliminary controlled study. AB - The aim of this pilot study was to compare cerebral changes on magnetic resonance imaging (MRI) scans in elderly schizophrenic subjects with those in psychiatric and normal control subjects. We compared the MRIs of 19 subjects with schizophrenia, 19 age- and gender-matched subjects with recurrent major depression, and 19 age- and gender-matched nonpsychiatric control subjects. Deep white matter hyperintensities (DWMH) in right posterior regions were significantly more prominent in the schizophrenic group than in the two comparison groups. Total ratings of MRI abnormalities were significantly related to age in both the normal control and schizophrenic groups, but not in the depressive group. Age of onset was positively associated with total ratings in the depressive group, but not in the schizophrenic group. Thus, a subset of elderly patients with schizophrenia appear to have cerebral white matter abnormalities; such abnormalities may not be confined to late-onset schizophrenia. Systematic MRI studies of early- and late-onset schizophrenia in late life are needed to resolve this question. PMID- 8723303 TI - Slow wave sleep and computed tomographic measures of brain morphology in schizophrenia. AB - To test the hypothesis that slow wave sleep in schizophrenia is inversely correlated with ventricular system volume, polysomnography and computed tomographic (CT) brain imaging were carried out in 14 psychiatric patients who met Research Diagnostic Criteria for schizophrenia (h = 11) or schizoaffective disorder (n = 3). Three measures of ventricular system volume were analyzed: (1) raw ventricular volume expressed in cm3; (2) ventricle-to-brain ratio; and (3) ventricular volume corrected for normal variation in age and head size expressed as a standardized (z) score. All three quantifications of ventricular volume were significantly and inversely correlated with visually scored measures of stage 3 and stage 4 sleep. This finding suggests that the etiology of slow wave sleep deficits in schizophrenia is related either directly or indirectly to underlying brain dysmorphology. PMID- 8723305 TI - A simple clinical assessment of attention in schizophrenia. AB - Although attentional dysfunction is considered a predominant feature of schizophrenia, it is seldom quantitatively assessed in clinical practice. A simple paper-and-pencil test of selective attention (Ruff et al., 1986) provides a 5-min assessment with theoretical relevance for schizophrenia. Thirty schizophrenic subjects performed the 2 and 7 Selective Attention Test, which measures speed and accuracy, and compares controlled and automatic information processing. Ninety percent of the subjects displayed automatic processing. Speed was severely impaired, and accuracy was less so. Sixty-seven percent of the schizophrenic subjects performed in the normal range (+/-1 SD) for accuracy, but only 23% scored in the normal range for speed. Speed and accuracy were moderately correlated. Although theories of a lateralized deficit were not supported, the results are consistent with a hypothesized frontal lobe dysfunction. The automatic processing demonstrated may represent old learning. Poor controlled processing may preclude the acquisition of automatic functioning. This selective attention test may provide a measure of severity of cognitive impairment relevant to daily life functioning. PMID- 8723304 TI - Assessing declarative memory in schizophrenia using Wisconsin Card Sorting Test stimuli: the Paired Associate Recognition Test. AB - The Paired Associate Recognition Test (PART) was developed to measure declarative memory using Wisconsin Card Sorting Test (WCST) stimuli, so that both tasks could be administered during functional neuroimaging to differentiate memory and executive function, and associated frontal and temporal lobe activation in schizophrenia. The current study was designed to compare PART and WCST performance in schizophrenic patients and to examine effects of medication and symptomatology. The PART, WCST, and standard declarative memory tasks were administered to 30 chronic schizophrenic patients and 30 matched healthy control subjects. Supporting task validity was the finding that patients were equally impaired on the PART and the WCST. Neuroleptics did not appear to affect performance. The effect of anticholinergic medication correlated negatively with WCST performance in a small subsample. Severity of schizophrenia-specific symptoms measured at intake on the Brief Psychiatric Rating Scale correlated negatively with performance on the WCST. These results support the application of the PART and WCST in future functional neuroimaging studies. PMID- 8723306 TI - Auditory sustained attention in schizophrenia: a comparison of the Continuous Performance Test and the Pigache Attention Task. AB - An auditory version of the Continuous Performance Test (CPT) was compared with the Pigache Attention Task (PAT) to evaluate relative sensitivities in schizophrenia. Digits spoken seamlessly for 5 min required a button-press response to each of 50 targets. Diotic and dichotic event rates were 1/s for the CPT and 0.5/s and 2/s for the PAT. 'Correct' responses were restricted to the interstimulus interval (CPT) or to longer intervals (PAT). This study demonstrated that the CPT method of scoring is invalid since significant numbers of late responses counted as both omission and commission errors. The CPT was significantly easier than the PAT, which provided a more appropriate range of difficulty. The PAT again discriminated between healthy subjects and schizophrenic patients. PMID- 8723307 TI - Effects of crack cocaine on neurocognitive function. AB - Because crack cocaine appears to have a preferential effect on the metabolic and electrophysiological activity of the frontal and temporal regions of the brain (Pascual-Leone et al., 1991a, 1991b; Volkow, 1992), we hypothesized that cognitive measures of those regions would be impaired in crack cocaine users relative to measures in normal volunteers. We used logistic regression to determine the relationship of cocaine usage to neuropsychological test performance. We compared 38 patients with an average of 3.6 (SD = 2.5) years of crack cocaine use and 24.5 (SD = 28.1) days of abstinence to 54 normal volunteers on a battery of neuropsychological tests. Statistical adjustments were made for the effects of age, education, socioeconomic class, and level of depression. Our findings were mixed with regard to purported measures of executive/frontal functioning, with worse performance associated with cocaine usage on the Booklet Categories Test, but better performance associated on others (number of categories on the Wisconsin Card Sorting Test, Controlled Oral Word Association). Cocaine usage was associated with impairment on measures of spatial, but not verbal memory, confrontation naming, and Trail-making Test, Part B, a measure of perceptual-motor speed and cognitive flexibility. In summary, it appears that continuous crack cocaine use produces a dissociative pattern in neuropsychological test performance with improvement on some measures, but deterioration on others. The permanence of these effects remains to be determined with longitudinal studies. PMID- 8723308 TI - Serotonin indices and impulsivity in normal volunteers. AB - Hormonal responses to oral paroxetine were examined in a group of healthy subjects. The calcium response to serotonin (5-hydroxytryptamine, 5HT), mediated by platelet 5HT2A, was also measured. Paroxetine elicited a cortisol response that was directly correlated with the magnitude of platelet calcium response. The cortisol response was also correlated with the trait of impulsivity. These results suggest that paroxetine may be a useful probe in studies of serotonergic systems. PMID- 8723309 TI - Association of levels of N-acetyl-beta-glucosaminidase with severity of psychiatric symptoms in panic disorder. AB - Urinary levels of N-acetyl-beta-glucosaminidase (NAG) were measured in 58 patients with panic disorder. NAG levels were found to be significantly related to the severity of 23 of 72 mood states, measured by the Profile of Mood States, which were grouped in three categories: hostility or irritability, sadness, and panic. A similar result was found in a previous study of bipolar patients. NAG levels were also related to scores on the Hamilton Rating Scale for Anxiety and the Sheehan Patient-Rated Anxiety Scale. It is speculated that NAG could be a marker for serotonin. PMID- 8723310 TI - Biological and clinical validation of atypical depression. AB - Depressed patients with (a) mood reactivity alone (MR group), (b) mood reactivity plus one or more associated features (atypical depression, AD group), and (c) patients with neither mood reactivity nor atypical depression (non-MR/AD group) were compared on their cortisol response to 75 mg of desipramine (DMI), a relatively selective norepinephrine reuptake inhibitor. AD patients exhibited a significantly higher cortisol response to DMI compared with MR and non-MR/AD patients, suggesting that atypical depression may be associated with a less impaired norepinephrine system. MR and non-MR/AD patients did not differ, suggesting that mood reactivity alone is not associated with the biological profile observed in atypical depression. Results indicate that while mood reactivity may be necessary for the diagnosis of atypical depression, the additional presence of at least one associated symptom is required for a distinct biological profile. Our findings provide further biological validation of the concept of atypical depression. PMID- 8723311 TI - Violence in hospitalized psychiatric patients: diurnal and seasonal patterns. AB - The use of four-point physical restraints on a single occasion or repeatedly in psychiatric inpatients was followed prospectively during 1993 in Beer Sheva Mental Health Center. Mood-disordered patients had significantly more restraints per patient than did schizophrenic and schizoaffective patients. The monthly rates of restraints of mood-disordered and schizophrenic patients were found to be negatively correlated with the duration of the daily photoperiod, which was phase-advanced by a month. The findings suggest that seasonality (the duration of the daily photoperiod) should be taken into account in attempts to predict the potential dangerousness of psychiatric patients. PMID- 8723312 TI - No season-of-birth effect in schizophrenic patients from a tropical island in the Southern Hemisphere. AB - The distribution of birth rates was examined in 668 schizophrenic patients born in Reunion, a tropical French island in the Southern Hemisphere, and compared with that in the general local population. We failed to observe a significant season-of-birth effect, either in the total sample of schizophrenic patients or in subgroups categorized by gender, age, or family history of schizophrenia. Seasonal factors do not appear to affect the yearly distribution of births among schizophrenic patients on Reunion Island. It is nevertheless possible that environmental factors which are seasonal in countries with more contrasting climates have a continuous effect, throughout the whole year, in subtropical areas. Conversely, these findings provide some evidence against the hypothesis that there is an age-incidence artifact in seasonal studies from countries in the Northern Hemisphere. PMID- 8723313 TI - Automated docking of flexible ligands: applications of AutoDock. AB - AutoDock is a suite of C programs used to predict the bound conformations of a small, flexible ligand to a macromolecular target of known structure. The technique combines simulated annealing for conformation searching with a rapid grid-based method of energy evaluation. This paper reviews recent applications of the technique and describes the enhancements included in the current release. PMID- 8723314 TI - The effect of water activity on the association constant and the enthalpy of reaction between lysozyme and the specific antibodies D1.3 and D44.1. AB - The reactions of lysozyme with the specific monoclonal antibody D1.3, its Fv fragment and a mutant of the Fv, were studied under conditions of reduced water activity through the addition of the cosolutes glycerol, ethanol, dioxane and methanol. Titration calorimetry, BIAcoreTM and ultracentrifugal analyses were used to determine enthalpy of reactions and affinity constants. There was a decrease in the values of the enthalpies of reactions as well as in the association constants which was proportional to the decrease in water activity. These results are consistent with a structural model in which water molecules bound to the antigen and the antibody are conserved upon complex formation and provide bonds which are important for the stability of the complex. In contrast, the reaction of lysozyme with the specific monoclonal antibody D44.1, or its Fab, showed the inverse effect: a small increase in the value of the association constant with decreasing water molarities. This is in agreement with a model in which binding of antigen to antibody D44.1 is accompanied by the release of a very small number of water molecules. PMID- 8723315 TI - Lac repressor-operator interaction: N-terminal peptide backbone 1H and 15N chemical shifts upon complex formation with DNA. AB - When the lac repressor tetramer is bound to its DNA operator, methylation protection shows the nearly symmetric operator half-sites are contracted asymmetrically. This asymmetric binding results from the DNA sequence/structure. The reported structure of lac repressor N-terminal fragment and an 11 base-pair operator left half-site provides no information concerning the effect of asymmetric binding, from left operator half-site to right half-site, upon the polypeptide backbone. We isolated uniformly 15N labeled 56 amino acid wild-type (HP56WT) and 64 residue mutant [Pro3 > Tyr3] (HP64tyr3) lac repressor N-terminal DNA binding fragments for 1H/15N NMR studies with the left and right operators separately. Spectral coincidence of these longer fragments, indicating structural similarity with a protease derived 51 amino acid fragment for which the amide correlations are assigned, allows for assignment of the common amide resonances. For both HP56WT and HP64tyr3, spectral overlap of the amide correlation peaks reveals the polypeptide backbones of the uncomplexed polypeptides are structurally similar. Likewise the complexes of the peptides to the 11 base-pair lac left operator half-site are similar. On the other hand, complexes of HP56WT and the left compared to the right lac operator half-site show different residues of the polypeptide are affected by binding different half-sites of the operator. Thus, the DNA sequence/structure transmits asymmetry to the polypeptide backbone of the interacting protein. PMID- 8723316 TI - Roles of lysine-69 in dimerization and activity of Trimeresurus flavoviridis venom aspartate-49-phospholipase A2. AB - Trimeresurus flavoviridis (Habu snake) venom aspartate-49-phospholipase A2 (Asp 49-PLA2) was reacted at pH 9.0 with a 2-fold molar excess of 2,4,6 trinitrobenzenesulfonate in the absence of Ca2+ and two trinitrophenylated derivatives were isolated by HPLC. One was a derivative modified at Lys-11 and its activity was mostly retained. The other was a derivative modified at both Lys 11 and Lys-72 and its activity was 40% that of unmodified enzyme. Trinitrophenylation of Lys-72 appeared to bring about a conformational disorder at the lipid-water interface recognition site and thus a reduction of activity. When the enzyme was modified in the presence of Ca2+, activity decreased at a rate much faster than that in the absence of Ca2+ and Lys-69 came to be modified. These results suggested that conformational displacement of Asp-49-PLA2 of a local to global type occurs upon the binding of Ca2+. The derivative modified at Lys-69 had 28% activity and existed as a monomer. This supports a previous assumption that Lys-69 participates in dimerization of group II Asp-49-PLA2s [Brunie et al. (1985) J. Biol. Chem. 260, 9742-9749] and shows that dimerization is not necessarily essential for activity manifestation. PMID- 8723317 TI - Binding of fibrinogen to platelet integrin alpha IIb beta 3 in solution as monitored by tracer sedimentation equilibrium. AB - Fibrinogen showed essentially no binding (KD > 1 mM) to platelet alpha IIb beta 3 integrin in solution in the presence of Triton or octylglucoside above critical micellar concentrations. Under these conditions the integrin was an alpha beta monomer. After removal of the detergent from the Triton containing buffer (25 mM Tris/HCl;, 150 mM NaCl, 1 mM CaCl2, 1 mM MgCl2, pH 7.4) the integrin formed aggregates with hexamers as the most prominent species, as demonstrated by analytical ultracentrifugation and electron microscopy. Tracer sedimentation equilibrium experiments indicate that fibrinogen binds to the integrin aggregates, but with a surprisingly large KD (at least 3 microM). This value is 10- to 100-fold higher than values determined by solid phase assays or with integrins reconstituted onto lipid bilayers. PMID- 8723318 TI - Comparative interaction kinetics of two recombinant Fabs and of the corresponding antibodies directed to the coat protein of tobacco mosaic virus. AB - Two recombinant Fab fragments, 57P and 174P, recognizing peptide 134-146 of the coat protein of tobacco mosaic virus have been cloned, sequenced and expressed in Escherichia coli. They differ by 15 amino acid changes in the sequence of their variable region. The interaction kinetics of the Fabs with the wild-type and four mutant peptides have been compared using a BIAcoreTM biosensor instrument. The recombinant Fab 174P had the same reactivity as the Fab fragment obtained by enzymatic cleavage of monoclonal antibody 174P. The two recombinant Fabs recognized the various peptides in the same ranking order but Fab 174P consistently dissociated somewhat faster from the peptides compared to Fab 57P. The two whole antibodies showed the same relative differences in reactivity as the two recombinant Fabs. The location of amino acid changes was visualized on a model structure of the Fab. Differences in dissociation rates of the two antibodies are most likely due to changes located at the periphery of the antigen combining site and/or at the interface between the light and heavy chain domains. Our results demonstrate the feasibility of detecting very small differences in binding affinity by the biosensor technology, which is a prerequisite for assessing the functional effect of limited structural changes. PMID- 8723319 TI - Metal-induced hepatotoxicity. AB - Figure 3 summarizes several proposed mechanisms of iron- or copper-induced hepatotoxicity. It has long been suspected that free radicals may play a role in iron- and copper-induced cell toxicity because of the powerful prooxidant action of iron and copper salts in vitro. In the presence of available cellular reductants, iron or copper in low molecular weight forms may play a catalytic role in the initiation of free radical reactions. The resulting oxyradicals have the potential to damage cellular lipids, nucleic acids, proteins, and carbohydrates, resulting in wide-ranging impairment in cellular function and integrity. However, cells are endowed with cytoprotective mechanisms (antioxidants, scavenging enzymes, repair processes) that act to counteract the effects of free radical production. Thus, the net effect of metal-induced free radicals on cellular function will depend on the balance between radical production and the cytoprotective systems As a result, there may be a rate of free radical production that must be exceeded before cellular injury occurs. Evidence has now accumulated that iron or copper overload in experimental animals can result in oxidative damage to lipids in vivo, once the concentration of the metal exceeds a threshold level. In the liver, this lipid peroxidation is associated with impairment of membrane-dependent functions of mitochondria (oxidative metabolism) and lysosomes (membrane integrity, fluidity, pH). Although these findings do not prove causality, it seems likely that lipid peroxidation is involved, since similar functional defects are produced by metal-induced lipid peroxidation in these organelles in vitro. Both iron and copper overload impair hepatic mitochondrial respiration, primarily through a decrease in cytochrome c oxidase activity. In iron overload, hepatocellular calcium homeostasis may be impaired through damage to mitochondrial and microsomal calcium sequestration. DNA has also been reported to be a target of metal-induced damage in the liver; this may have consequences as regards malignant transformation. The levels of some antioxidants in the liver are decreased in rats with iron or copper overload, which is also suggestive of ongoing oxidative stress. Reduced cellular ATP levels, lysosomal fragility, impaired cellular calcium homeostasis, and damage to DNA may all contribute to hepatocellular injury in iron and copper overload. There are few data addressing the key issue of whether free radical production is increased in patients with iron or copper overload. Patients with hereditary hemochromatosis have elevated plasma levels of TBA-reactants and increased hepatic levels of MDA-protein and HNE-protein adducts, indicative of lipid peroxidation. Mitochondria isolated from the livers of Wilson disease patients have evidence of lipid peroxidation, and some patients with Wilson disease have decreased hepatic and plasma levels of vitamin E. Additional investigation will be required to fully assess oxidant stress and its potential pathophysiologic role in patients with iron or copper overload. PMID- 8723320 TI - Metals, oxidative stress, and hepatic fibrogenesis. PMID- 8723321 TI - The role of metals in ischemia/reperfusion injury of the liver. AB - Based on current information, we have described the role that metals play in potentiating and ameliorating liver I/R injury. To date, most of the data have focused on the deleterious effects of free iron in mediating I/R injury. Several therapeutic strategies have proven useful in animal models to counteract the effect of iron as a potentiator of I/R injury. These approaches have predominantly centered on the role of iron chelation using DFO and DFO conjugates. The data suggest that chelation of iron may prove useful in preventing I/R injury such as occurs in liver transplantation. Indeed, enough data are now available to initiate and support clinical trials (e.g., addition of DFO conjugates to explant storage solutions). The role of copper, however, is less well defined. Copper is important for the function of copper-zinc SOD. However, free copper may be as injurious as free iron. Further studies are needed to clarify the role of copper in I/R-induced hepatocellular necrosis. Selenium has a well-defined antioxidant role as part of GSH peroxidase (GSH antioxidant pathway). More recent data suggest that selenium may also act as an antioxidant through selenoprotein P, but the role of selenoprotein P in I/R injury remains to be defined. Finally, zinc appears to function as an antioxidant in less well defined pathways. Further studies are needed to identify the fundamental mechanisms by which zinc may ameliorate oxidative damage during I/R injury. These data demonstrate that metals play a critical role in I/R injury of the liver and remain a fruitful area for investigation and development of therapeutic strategies. PMID- 8723322 TI - Antioxidant defenses in metal-induced liver damage. AB - Recent investigations have begun to define more clearly the cellular and molecular roles of oxidant stress in mediating the liver injury and fibrosis of metal storage diseases. Because of a variety of perturbations in antioxidant homeostasis in iron and copper overload, restoring the antioxidant balance to normal, or even exceeding normal levels of selected antioxidants, may provide additional protection against liver injury and prevent the progression to fibrosis and cirrhosis. Inasmuch as GSH levels appear to be elevated in livers of experimentally iron-overloaded animals, attempts to increase this antioxidant should perhaps be limited to copper overload conditions in which hepatic GSH is low. Vitamin C (ascorbate) supplementation should probably be avoided in all metal overload states because of its potentiation of radical generation by transition metals. The safety of beta-carotene in alcoholic liver disease has been questioned. Therefore, until more is known about its toxicity in metal overload, beta-carotene may not be an ideal antioxidant for clinical trials. Vitamin E and related compounds, therefore, appear to be the most reasonable antioxidants to test in metal overload states at this time. In the near future, the results of controlled clinical trials of the use of antioxidants in these and other liver disorders will hopefully provide clearer guidelines for their safety and possible use. PMID- 8723323 TI - Evolving expression of hereditary hemochromatosis. PMID- 8723324 TI - Hemochromatosis: genetics and pathogenesis. PMID- 8723325 TI - Iron in liver diseases other than hemochromatosis. AB - There is growing evidence that normal or only mildly increased amounts of iron in the liver can be damaging, particularly when they are combined with other hepatotoxic factors such as alcohol, porphyrogenic drugs, or chronic viral hepatitis. Iron enhances the pathogenicity of microorganisms, adversely affects the function of macrophages and lymphocytes, and enhances fibrogenic pathways, all of which may increase hepatic injury due to iron itself or to iron and other factors. Iron may also be a co-carcinogen or promoter of hepatocellular carcinoma, even in patients without HC or cirrhosis. Based on this and other evidence, we hope that the era of indiscriminate iron supplementation will come to an end. Bloodletting, a therapy much in vogue 2 centuries ago, is deservedly enjoying a renaissance, based on our current understanding of the toxic effects of iron and the benefits of its depletion. PMID- 8723326 TI - Wilson disease: genetic basis of copper toxicity and natural history. AB - The discovery that the gene for Wilson disease encodes a copper-transporting ATPase has greatly improved our understanding of the pathophysiology of this disorder and of copper metabolism in humans. The abundance of disease-specific mutations and their location at multiple sites across the genome have limited molecular genetic diagnosis to kindred of known patients, and confirm the necessity for de novo screening by well-proven clinical and biochemical means. It is uncertain whether the variety of specific mutations will account for the wide range of presenting clinical signs and symptoms of Wilson disease, and environmental and extragenic factors are likely to be important contributing factors. Chelation therapy with penicillamine and trientine remain effective treatment for most symptomatic hepatic and neurologic Wilson disease. Zinc salts may be used for some asymptomatic patients, and OLT for fulminant hepatitis and patients for whom pharmacotherapy is ineffective. The chelating agent tetrathiomolybdate is under investigation for the treatment of neurologic Wilson disease. Gene therapy is the new horizon for treatment of Wilson disease. However, the ability to treat this disorder effectively by this means awaits further characterization of the gene product and more efficient methods for gene delivery to all hepatocytes in the liver. PMID- 8723327 TI - A 49-year-old man with alpha 1-antitrypsin deficiency and abnormal iron study results. PMID- 8723328 TI - The mysterious mechanism of growth. PMID- 8723329 TI - Enteropneusts and chordate evolution. PMID- 8723330 TI - Population genetics. PMID- 8723331 TI - Visual attention: spotlights, highlights and visual awareness. PMID- 8723332 TI - Cytoskeleton: a catastrophic kinesin. AB - The 'plus' ends of microtubules exhibit dynamic instability, switching stochastically from growth to shortening phases. The first endogenous regulator of such 'catastrophes' has been identified, and is a kinesin-related microtubule motor protein. PMID- 8723333 TI - Craniofacial development: knockout mice take one on the chin. PMID- 8723334 TI - Basal ganglia: new therapeutic approaches to Parkinson's disease. PMID- 8723335 TI - Transcriptional control: how nuclear receptors get turned on. AB - The recently determined structures of the ligand-binding domains from three nuclear receptors show that a receptor undergoes a significant conformational change on ligand binding. It is not yet clear how this structural change results in transcriptional activation. PMID- 8723336 TI - Synaptic plasticity: the basis of particular types of learning. AB - A new twist to the relationship between synaptic plasticity and learning and memory is revealed by the latest series of studies comparing animal behavior with electrophysiological recordings in the hippocampus. PMID- 8723337 TI - Drosophila morphogenesis: follow-my-leader in epithelia. AB - The shape of an organism largely depends on movements of epithelial sheets during embryogenesis. The discovery of a second MAPK kinase in Drosophila provides new insights into mechanisms that control the coordinated movement of epithelial cells. PMID- 8723338 TI - Cell signalling: MAGUK magic. AB - Diverse extracellular signals are coupled at the plasma membrane to intracellular signal transduction pathways and the cytoskeleton. Members of a ubiquitous multidomain family of proteins, the MAGUK proteins, are emerging as common mediators of this coupling. PMID- 8723339 TI - Selfish DNA: the best defense is a good offense. AB - The recent discovery of novel biochemical activities of intron-encoded endonucleases emphasizes the selfish nature of mobile genetic elements. PMID- 8723340 TI - Perception: the seeing ear. AB - Some people perceive colours when they hear words--a phenomenon known as synaesthesia. Brain imaging techniques have revealed the areas of the cortex involved in this rare mingling of the senses. PMID- 8723341 TI - Molecular motors: single-molecule mechanics. AB - Novel techniques are revealing the movements and forces associated with single interactions of motor proteins, such as myosin and kinesin, and also of processive enzymes, such as RNA polymerase. PMID- 8723342 TI - Neurophysiology: the changing face of inhibition. AB - Inhibition is commonly thought to suppress neuronal responses, but new discoveries suggest that it may also gate transmission by coordinating the temporal patterning of neuronal responses and so play an important part in information processing in the brain. PMID- 8723343 TI - Lymphocyte activation: T-cell regulation by CTLA-4. PMID- 8723344 TI - Transcription: building an initiation machine. AB - Recent crystal and solution structures of components of the core transcription initiation complex that assembles at RNA polymerase II promoters reveal similarities to core histones and cyclin A. PMID- 8723345 TI - Sequencing and analysis of bacterial genomes. AB - The complete sequences of two small bacterial genomes have recently become available, and those of several more species should follow within the next two years. Sequence comparisons show that the most bacterial proteins are highly conserved in evolution, allowing predictions to be made about the functions of most products of an uncharacterized genome. Bacterial genomes differ vastly in their gene repertoires. Although genes for components of the translation and transcription machinery, and for molecular chaperones, are typically maintained, many regulatory and metabolic systems are absent in bacteria with small genomes. Mycoplasma genitalium, with the smallest known genome of any cellular life form, lacks virtually all known regulatory genes, and its gene expression may be regulated differently than in other bacteria. Genome organization is evolutionarily labile: extensive gene shuffling leaves only very few conserved gene arrays in distantly related bacteria. PMID- 8723346 TI - Vitamin A-deficient quail embryos have half a hindbrain and other neural defects. AB - BACKGROUND: Retinoic acid (RA) is a morphogenetically active signalling molecule thought to be involved in the development of severely embryonic systems (based on its effect when applied in excess and the fact that it can be detected endogenously in embryos). Here, we adopt a novel approach and use the vitamin A deficient (A-) quail embryo to ask what defects these embryos show when they develop in the absence of RA, with particular reference to the nervous system. RESULTS: We have examined the anatomy, the expression domains of a variety of genes and the immunoreactivity to several antibodies in these A- embryos. In addition to the previously documented cardiovascular abnormalities, we find that the somites are smaller in A- embryos, otic vesicle development is abnormal and the somites continue up to and underneath the otic vesicle. In the central nervous system, we find that neural crest cells need RA for normal development and survival, and the neural tube fails to extend any neurites into the periphery. Using general hindbrain morphology and the expression patterns of Hoxa 2, Hoxb-1, Hoxb-4, Krox-20 and FGF-3 as markers, we conclude that segmentation in the myelencephalon (rhombomeres 4-8) is disrupted. In contrast, the dorsoventral axis of the neural tube using Shh, islet-1 and Pax-3 as markers is normal. CONCLUSIONS: These results demonstrate at least three roles for RA in central nervous system development: neural crest survival, neurite outgrowth and hindbrain patterning. PMID- 8723347 TI - Calcium-mediated apoptosis in a plant hypersensitive disease resistance response. AB - BACKGROUND: Avirulent pathogens elicit a battery of plant defenses, often accompanied by collapse of the challenged cells. In soybean cells, sustained accumulation of H2O2 from an oxidative burst cues localized host cell death. Such hypersensitive cell death appears to be an active process, but little is known about the mechanisms underlying cellular collapse. RESULTS: We show that H2O2 stimulates a rapid influx of Ca2+ into soybean cells, which activates a physiological cell death program resulting in the generation of large (approximately 50 kb) DNA fragments and cell corpse morphology--including cell shrinkage, plasma membrane blebbing and nuclear condensation--characteristic of apoptosis. In contrast, H2O2 induction of the cellular protectant gene glutathione S-transferase is Ca(2+)-independent. Apoptosis in soybean cells and leaf tissue was induced by avirulent Pseudomonas syringae pv. glycinea but was not observed at comparable stages of the compatible interaction with the isogenic virulent strain, which fails to elicit a hypersensitive response. Apoptosis was also observed at the onset of the hypersensitive response in Arabidopsis leaves inoculated with avirulent P. syringae pv. tomato and in tobacco cells treated with the fungal peptide cryptogein, which is involved in the induction of non host resistance to Phytophthora cryptogea. CONCLUSIONS: These observations establish a signal function for Ca2+ downstream of the oxidative burst in the activation of a physiological cell death program in soybean cells that is similar to apoptosis in animals. That the characteristic cell corpse morphology is also induced in Arabidopsis and tobacco by different avirulence signals suggests that apoptosis may prove to be a common, but not necessarily ubiquitous, feature of incompatible plant-pathogen interactions. Emerging similarities between facets of hypersensitive disease resistance and the mammalian native immune system indicate that apoptosis is a widespread defence mechanism in eukaryotes. PMID- 8723348 TI - Multiple forms of an inositol polyphosphate 5-phosphatase form signaling complexes with Shc and Grb2. AB - BACKGROUND: Shc and Grb2 form a complex in cells in response to growth factor stimulation and link tyrosine kinases to Ras during the resulting signaling process. Shc and Grb2 each contain domains that mediate interactions with other unidentified intracellular proteins. For example, the Shc PTB domain binds to 130 kDa and 145 kDa tyrosine-phosphorylated proteins in response to stimulation of cells by growth factors, cytokines and crosslinking of antigen receptors. The Grb2 SH3 domains bind to an unidentified 116 kDa protein in T cells. We have identified three proteins, of 110 kDa, 130 kDa and 145 kDa, as a new family of molecules encoded by the same gene. In vivo studies show that these proteins form signal transduction complexes with Shc and with Grb2. RESULTS: The 130 kDa and 145 kDa tyrosine-phosphorylated proteins that associate with the Shc PTB domain were purified by conventional chromatographic methods. Partial peptide and cDNA sequences corresponding to these proteins, termed SIP-145 and SIP-130 (SIP for signaling inositol polyphosphate 5-phosphatase), identified them as SH2 domain containing products of a single gene and as members of the inositol polyphosphate 5-phosphatase family. The SIP-130 and SIP-145 proteins and inositol polyphosphate 5-phosphatase activity associated with Shc in vivo in response to B-cell activation. By using an independent approach, expression cloning, we found that the Grb2 SH3 domains bind specifically to SIP-110, a 110 kDa splice variant of SIP-145 and SIP-130, which lacks the SH2 domain. The SIP proteins hydrolyzed phosphatidylinositol (3,4,5)-trisphosphate (PtdIns (3,4,5)-P3) and Ins (1,3,4,5) P4, but not PtdIns (4,5)-P2 or Ins (1,4,5)-P3. CONCLUSIONS: These findings strongly implicate the inositol polyphosphate 5-phosphatases in Shc- and Grb2 mediated signal transduction. Furthermore, SIP-110, SIP-130 and SIP-145 prefer 3 phosphorylated substrates, suggesting a link to the phosphatidylinositol 3-kinase signaling pathway. PMID- 8723349 TI - A mechanism of Bud1p GTPase action suggested by mutational analysis and immunolocalization. AB - BACKGROUND: Yeast cells polarize, bud, and divide in either of two genetically programmed patterns: axial or bipolar. The Saccharomyces cerevisiae gene BUD1 (also known as RSR1) encodes a Ras-related GTPase critical for selection of a bud sites in these patterns. To distinguish between possible mechanisms of Bud1p action, we have examined the function and subcellular localization of Bud1p in a variety of mutant situations. RESULTS: Bud1p has 57% identity to H-ras, except for an 81 amino-acid insertion near the carboxyl terminus. Mutation of the proposed BUD1 effector domain produces a protein which can neither support normal patterns of budding nor interact with CDC24, which encodes a likely Bud1p effector. A version of Bud1p deleted for the 81 amino-acid unique region is essentially wild-type. Immunofluorescence and cell fractionation indicate that Bud1p remains associated with the membrane throughout its GTPase cycle. Both potential effectors of Bud1p, Bem1p and Cdc24p, are also membrane associated even in the absence of Bud1p, suggesting that Bud1p is not required to dock these proteins from the cytosol but may couple these proteins and others within the plane of the plasma membrane. CONCLUSIONS: Based upon observations reported here and elsewhere, we propose a novel mechanism of Bud1p GTPase action. Like Ras, Bud1p GTPase is constitutively associated with the plasma membrane; however, concentrated activities of Bud5p GDP-GTP exchange factor and Bud2p GTPase activating protein at the future bud site promote rapid cycling of Bud1p between GTP- and GDP-bound conformations in a spatially restricted manner. Local GTPase cycling serves to efficiently nucleate complexes between polarity establishment functions that direct cytoskeletal polarization towards the bud site. PMID- 8723350 TI - Identification of a novel ubiquitin-conjugating enzyme involved in mitotic cyclin degradation. AB - BACKGROUND: The destruction of cyclin B is required for exit from mitosis, and is mediated by the ubiquitin pathway. Recently, a 20S complex, termed the anaphase promoting complex (APC) or the cyclosome, has been genetically and biochemically identified as the cyclin-specific ubiquitin ligase (E3). In addition, a ubiquitin conjugating enzyme (E2), UBC4, was shown to be involved in cyclin ubiquitination in Xenopus egg extracts. Another E2 activity, designated UBCx, can independently support cyclin ubiquitination in Xenopus. A similar activity (E2-C) has also been observed in clams. However, the molecular identity of Xenopus UBCx or clam E2-C has not been established. RESULTS: We have purified and cloned Xenopus UBCx. Sequence comparisons with known E2s reveal that UBCx is a novel ubiquitin conjugating enzyme. Purified recombinant UBCx is sufficient to complement purified APC and E1 in destruction box-dependent cyclin ubiquitination. UBCx and UBC4 are active in a similar concentration range and with similar kinetics. At saturating enzyme concentrations, UBCx converts twice as much substrate into ubiquitin conjugates, but generates conjugates of lower molecular mass than UBC4. CONCLUSIONS: UBCx is a novel ubiquitin-conjugating enzyme involved in cyclin ubiquitination in Xenopus. Like UBC4, ubiquitination catalyzed by UBCx is dependent on both the destruction box and the APC, suggesting that these E2s function through a similar mechanism. However, as the patterns of conjugates generated by these E2s are distinct, these enzymes may play different roles in promoting cyclin proteolysis in mitosis. PMID- 8723351 TI - Large, complex modular structure of a fission yeast DNA replication origin. AB - BACKGROUND: In the budding yeast, Saccharomyces cerevisiae, each DNA replication origin is associated with an autonomously replicating sequence (ARS) element. Each element contains several modules, including an essential close match to the 11 base-pair (bp) ARS consensus sequence (ACS) and two or three short (< 20 bp) stimulatory motifs, within a stretch of approximately 150 bp or less. To determine whether a similar origin structure exists in the evolutionarily distant fission yeast, Schizosaccharomyces pombe, we used deletion and linker substitution scanning to identify the sequences important for the function of ars3002, a chromosomal replication origin. RESULTS: We detected two large (30-55 bp) essential regions and several additional stimulatory sequences within a 600 bp stretch of a restriction fragment containing ars3002. The two essential regions are similar to each other, and sequences similar to them are found in all known S. pombe ARS elements, suggesting that one or both of them may represent the S. pombe equivalent of the S. cerevisiae ACS. CONCLUSIONS: Like S. cerevisiae origins, the S. pombe origin, ars3002, possesses a modular structure, but the number and size of modules is greater for ars3002, and ars3002 is larger than S. cerevisiae origins. These observations suggest that origin function in S. pombe requires more protein-DNA interactions than in S. cerevisiae. PMID- 8723352 TI - E2F-1 but not E2F-4 can overcome p16-induced G1 cell-cycle arrest. AB - BACKGROUND: The transition from G1 to S phase is the key regulatory step in the mammalian cell cycle. This transition is regulated positively by G1-specific cyclin-dependent kinases (cdks) and negatively by the product of the retinoblastoma tumour suppressor gene, pRb. Hypophosphorylated pRb binds to and inactivates the E2F transcription factor, which controls the expression of genes required for S-phase progression. Hyperphosphorylation of pRb in late G1 phase results in the accumulation of active E2F, a critical event in the progression to S phase. The E2F factor is not a single entity, but rather represents a family of highly related molecules, all of which bind to pRb or the pRb-related proteins p107 and p130. RESULTS: In this study, we have used specific inhibitors of cdks to explore the requirements for cell-cycle progression from G1 to S phase. Expression of p16Ink4, which specifically inhibits cyclin D-directed cdks, blocks cells in G1 phase; this block can be overcome by expression of the viral proteins that inactivate pRb or by E2F-1. Importantly however, the G1 arrest is not overcome by overexpression of E2F-4. By using chimeric E2F proteins, containing amino-acid sequences from E2F-1 and E2F-4, we have shown that their differential abilities to overcome a p16-imposed arrest is determined by their respective amino-terminal regions. We also demonstrate that E2F-1 can promote entry into S phase without concomitant phosphorylation of pRb. In contrast to the p16-mediated G1 block, G1 arrest mediated by the cdk inhibitors p21Cip1 or p27Kip1 cannot be bypassed either by inactivation of pRb or overexpression of E2F family members. CONCLUSIONS: These data demonstrate that the role of the cyclin D-directed cdks in promoting the progression of cells from G1 into S phase is wholly to activate an E2F-1-like activity through phosphorylation, thus preventing the formation of the E2F-pRb complex. The cyclin E-cdk2 complex is also required for the G1/S transition but has a different and as yet undefined role. We also provide evidence for a functional difference between E2F-1 and E2F-4, dependant upon the region that contains the DNA-binding and dimerization domains. These results indicate that these two E2F family members are likely to regulate the expression of different subsets of E2F-responsive promoters. PMID- 8723353 TI - MSH6, a Saccharomyces cerevisiae protein that binds to mismatches as a heterodimer with MSH2. AB - The process of post-replicative DNA-mismatch repair seems to be highly evolutionarily conserved. In Escherichia coli, DNA mismatches are recognized by the MutS protein. Homologues of the E. coli mutS and mutL mismatch-repair genes have been identified in other prokaryotes, as well as in yeast and mammals. Recombinant Saccharomyces cerevisiae MSH2 (MSH for MutS homologue) and human hMSH2 proteins have been shown to bind to mismatch-containing DNA in vitro. However, the physiological role of hMSH2 is unclear, as shown by the recent finding that the mismatch-binding factor hMutS alpha isolated from extracts of human cells is a heterodimer of hMSH2 and another member of the MSH family, GTBP. It has been reported that S. cerevisiae possesses a mismatch-binding activity, which most probably contains MSH2. We show here that, as in human cells, the S. cerevisiae binding factor is composed of MSH2 and a new functional MutS homologue, MSH6, identified by its homology to GTBP. PMID- 8723354 TI - The procurement of vital organs: a synopsis of policy from various nations and the ethical implications of policy options. PMID- 8723355 TI - Comparison of the effects of a low-protein diet with the effects of a converting enzyme inhibitor on the progression of renal insufficiency in hypertensive rats. AB - The beneficial effects of a low-protein diet vs. angiotensin-converting enzyme inhibitor (ACEI, enalapril) on the course of ablation nephropathy (5/6 nephrectomy, 5/6NX) were compared in a new strain of genetic hypertensive rats, the Prague hypertensive rat (PHR). Both maneuvers were followed by a significant drop in proteinuria (1.27 and 8.8 vs. 56.2 mg/24 hod, p < 0.001, low-protein diet vs. ACEI vs. untreated), plasma levels of creatinine (175.3 and 177.1 vs. 245.3 mumol/L, p < 0.001) and urea (7.95 and 13.51 vs. 37.6 mmol/L, p < 0.001). Endogenous creatinine clearance was higher after both low-protein diet and ACEI than without them (134.6 and 127.8 vs. 56.7 microL/min/100 g BW, p < 0.001). Both maneuvers had a similar beneficial effect: no additional amelioration was observed with a combination of both low-protein diet and ACEI. Compared to normotensive Wistar rats, the results were quite similar in PHR except the blood pressure values; hypertension had no substantial effect on the course of 5/6NX or on the beneficial action of both low-protein diet and ACEI. PMID- 8723356 TI - Time course analysis of serum and urinary proteins by SDS-PAGE in experimental nephrotic syndrome. AB - Serum and urinary proteins from rats with nephrotic syndrome (NS) induced by puromycin aminonucleoside (PAN) were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Analysis was made on days 2, 4, 6, 8, 10, 12, 16, 20, and 30 after PAN injection. Data were compared with control rats (C). Rats developed proteinuria on days 4-30 and hypoproteinemia on days 4 16. Total protein concentration in serum and urine was similar on day 6. SDS-PAGE revealed that urinary albumin augmented on days 4-30 and serum albumin decreased markedly on days 4-20. Albumin concentration in serum and urine was similar on days 4-16. In addition, the study examined serum changes of 7 other proteins (designed as A, B, C, D, E, F, and G) which appeared or increased in urine, and whose molecular weights were higher (A, B, and C) or lower (D, E, F, and G) than that of albumin. In serum, protein A remained unchanged; protein B and G increased; proteins C, D, E, and F decreased. The qualitative pattern of urinary proteins remained essentially unchanged on days 4-30. During the intense proteinuria, the serum concentrations of protein B and albumin were similar and the urine concentrations of proteins C and D became comparable to that found in serum. These 7 serum proteins did not show the same behavior although all of them were excreted in urine. These data indicate that in PAN-nephrotic rats: (a) urinary proteins can be of low and high molecular weight, (b) serum proteins can be regulated independently of their urinary excretion and molecular weight, (c) the urine concentration of total protein and some specific proteins can reach values similar to that found in serum during the intense hypoproteinemia, and (d) the qualitative pattern of urinary proteins was unrelated to the magnitude of proteinuria. PMID- 8723357 TI - The influence of pefloxacine on experimental adriamycin-induced nephrotic syndrome in rats. AB - Initial reports on antiproteinuric effect of pefloxacine in small groups of patients with minimal-change nephropathy (MCN) and focal and segmental glomerulosclerosis (FSGS) have not been confirmed in other papers. To assess its antiproteinuric effect in experimental animals we administered pefloxacine to rats with adriamycin nephropathy showing morphological changes resembling human minimal-change disease or focal segmental glomerulosclerosis, and clinically with full-blown nephrotic syndrome. Pefloxacine treatment was at least partially effective in preventing further increase of proteinuria in rats with adriamycin nephropathy. The mechanism of this effect remains unclear and deserves further studies concentrating on the glomerular cytokine network and glomerular production of reactive oxygen species. PMID- 8723358 TI - Increased urinary trypsin-inhibitory activity in mercuric chloride induced nephrotoxicity in Wistar rats. AB - The relationship between trypsin-inhibitory activity (TIA) and the nephrotoxic effects of mercuric chloride (HgCl2)--as illustrated by proteinuria and by a drop in the glomerular filtration rate (GFR) measured by creatinine clearance test (CCT)--was investigated in Wistar rats. HgCl2, 150 or 250 micrograms/100 g BW per day was injected intraperitoneally three times a week for 2 weeks. Both groups showed a significant degree of proteinuria and urinary TIA. Group B (250 micrograms HgCl2/100 g BW) displayed a greater drop in GFR than group A (150 micrograms HgCl2/100 g BW). The urinary TIA was significantly correlated with proteinuria (group A: r = 0.87, group B: r = 0.84), but it was also significantly inversely correlated with the CCT (A: r = -0.96; B: r = -0.88). IN CONCLUSION: these results suggest that increased urinary TIA may be involved in and indicative of the pathogenesis of mercuric chloride induced nephrotoxicity. PMID- 8723359 TI - Ischemia-reperfusion injury of rat kidney relates more to tubular than to microcirculatory disturbances. AB - Several pathophysiological mechanisms have been purported to be involved in the development of acute ischemic renal failure, such as impairment of tubular function and/or of the renal microcirculation. However, it has not been elucidated as yet which of these mechanisms relates to the extent of kidney damage. Besides, little is known about the time course relationship between tubular and microcirculatory disturbances during the development of ischemia reperfusion injury. We therefore performed intravital videomicroscopy of the proximal tubules as well as the peritubular microcirculation of the rat renal cortex during the first 24 hr of reperfusion after varying lengths of warm ischemia (30 min, 30 WI group; 60 min, 60 WI group; 90 min, 90 WI group). In a separate group of animals subjected to the same protocol, the survival rate (SR) was determined. The SR in these groups were 100%, 20% and 0%, respectively. Initially, the tubular and microcirculatory changes (i.e., increased tubular diameter and reduced capillary blood flow) relate well to the length of warm ischemia as well as the SR. At a later stage of reperfusion, however, we observed that peritubular capillary blood flow and tubular diameter recovered more quickly in the 90 WI group than in the 30 WI and 60 WI groups. As a result, these parameters as obtained at 24 hr of reperfusion did not relate anymore to the survival rate. Besides, at this stage a severe loss of integrity of the tubular wall was noted in the 60 WI and 90 WI groups. These findings suggest that kidney viability is not determined by the extent of recovery of microcirculatory blood flow and/or tubular diameter during early reperfusion, but by the integrity of the tubular wall. PMID- 8723360 TI - Protective effect of N-benzoyl-beta-alanine against cisplatin nephrotoxicity in rats. AB - Prophylactic effects of N-benzoyl-beta-alanine (betamipron, BP), one of a series of N-acyl amino acids, were examined against cisplatin-induced nephrotoxicity. Male Wistar rats were injected i.p. with 6 mg/kg of cisplatin combined with an i.p. BP dose given at various times and various doses. Rats were sacrificed 5 days after cisplatin injection to weigh the kidney and liver, and to determine blood urea nitrogen (BUN) and serum creatinine (serum Cr) levels. Preliminary results suggest that treatment with BP is an effective means of protection against cisplatin-induced nephrotoxicity. Combination with BP reduced the weight loss following treatment with cisplatin. The ratios of the kidney and liver weights to the body weight in the animals treated with cisplatin followed later with BP are significantly different (p < 0.05) from those in the animals that received only cisplatin. The BUN and serum Cr levels in the animals treated with cisplatin followed from -1 to 4 hr, and from -4 to 4 hr later with 250 mg/kg BP dose and followed 1 hr later with from 250 to 1000 mg/kg, and from 250 to 2000 mg/kg BP doses differed significantly (p < 0.05) from those in the animals that received only cisplatin. Histological analysis of the kidneys confirmed the protective effect of BP. PMID- 8723361 TI - Hypercalcemia reduces renal medullary content of organic osmolytes. AB - Hypercalcemia is often associated with a urinary concentration defect. During antidiuresis, organic osmolytes [sorbitol, myo-inositol, taurine, and glycerophosphorylcholine (GPC)] accumulate in the renal inner medulla and are essential for urinary concentration. To clarify the relationship between organic osmolytes and urinary concentration defect in hypercalcemia, examination was made of the effects of hypercalcemia on renal medullary osmolytes content. Rats were put in a state of hypercalcemia by a calcium-rich diet supplemented with CaCO3 (2.5%/wt) and daily s.c. injection of 1.25(OH)2VitD3 (1.6 micrograms/kg). They were killed on days 7 and 14. Hypercalcemia induced a urinary concentration defect. Myo-inositol, sorbitol, and GPC contents in the renal medulla were significantly reduced. Aldose reductase activity decreased significantly. Hypercalcemia would thus appear to directly affect renal medullary content of organic osmolytes, thereby modifying renal concentration ability. PMID- 8723362 TI - Erythrocyte nucleotides and blood hypoxanthine in patients with uremia evaluated immediately and 24 hours after hemodialysis. AB - Using high-performance liquid chromatography, concentrations of erythrocyte adenine nucleotides and hypoxanthine were evaluated in patients undergoing regular acetate hemodialysis before dialysis, immediately following dialysis, and 24 hr after. It was shown that adenosine triphosphate concentration was maintained consistently high, not only just after hemodialysis but also 24 hr later. There was also no difference in concentration of mono- and diphosphates of adenosine. Hypoxanthine concentration decreased twofold after hemodialysis. However, it was still markedly higher than normal values. The level of hypoxanthine was maintained at the postdialysis level, 24 hr later. This suggests that hypoxanthine production could be stimulated during acetate dialysis. PMID- 8723363 TI - Midodrine hydrochloride in patients on hemodialysis with chronic hypotension. AB - Chronic hypotension, volume nonresponsive, is not rare in hemodialysis patients and is usually refractory to various therapies. Midodrine hydrochloride is an alpha-mimetic drug acting directly on the peripheral alpha-receptor and increases blood pressure. We studied 10 uremic patients on hemodialysis with chronic hypotension to evaluate efficacy and safety of midodrine. Midodrine hydrochloride was administered at a dose of 2.5 mg twice on the dialytic day, 1.25 mg twice on the nondialytic day. Subjective symptoms and objective parameters were evaluated and compared before and after midodrine treatment. Midodrine significantly increased systolic pressure from 73.0 +/- 10.5 mm Hg to 90.5 +/- 12.3 mm Hg (p < 0.01); and diastolic blood pressure from 44.0 +/- 8.4 mm Hg to 55.4 +/- 7.9 mm Hg (p < 0.01) before dialysis. Orthostasis, dizziness, fatigue, blurred vision, dullness, headache, and depression improved an average of 62%. All patients tolerated midodrine treatment well. Only mild side effects were noted, including flush sensation, neck soreness, and headache. We conclude that midodrine may be another choice for uremic patients on hemodialysis with chronic hypotension which responds poorly to other conventional management. PMID- 8723364 TI - Changes of plasma atrial and brain natriuretic peptide levels during hemodialysis. AB - OBJECTIVES: The purpose of this study was to evaluate the effect of hemodialysis on the plasma concentration of atrial and brain natriuretic peptides, and to determine the two-dimensional echocardiographic parameters affecting the changes of plasma atrial and brain natriuretic peptide levels in patients with chronic renal failure. BACKGROUND: Brain natriuretic peptide has been found in human cardiac tissue and increases in patients with congestive heart failure. However, the factors that stimulate the secretion of plasma brain natriuretic peptide have not yet been fully clarified. METHODS: In 15 patients with chronic renal failure, plasma atrial and brain natriuretic peptide levels and two-dimensional echocardiographic parameters were measured before and after each session of hemodialysis. RESULTS: Plasma atrial natriuretic peptide levels significantly decreased from 367 +/- 537 pg/mL to 138 +/- 167 pg/mL after hemodialysis (p < 0.01). However, plasma brain natriuretic peptide levels did not significantly change after hemodialysis. Left atrial dimension significantly decreased (41.1 +/ 6.6 vs. 36.3 +/- 6.2 mm, p < 0.01) and left ventricular end-diastolic dimension slightly decreased after hemodialysis (57.0 +/- 10.3 vs. 55.7 +/- 9.9 mm, p < 0.05). The decrease of left atrial dimension was greater than that of left ventricular end-diastolic dimension (4.9 +/- 1.6 vs. 1.3 +/- 0.6 mm, p < 0.05). Plasma brain natriuretic peptide levels significantly correlated with fractional shortening both before and after hemodialysis (r = 0.65, p < 0.05). CONCLUSION: Plasma atrial natriuretic peptide levels significantly decreased as the right and left atrial overloads decreased, and plasma brain natriuretic peptide levels did not significantly decrease after hemodialysis. Plasma brain natriuretic peptide levels were not significantly influenced by acute hemodynamic change, such as hemodialysis. However, plasma brain natriuretic peptide levels were significantly correlated with basic cardiac function. PMID- 8723365 TI - Possible relationship between hyperinsulinemia and glomerular hypertrophy in nephrosclerosis. AB - Hyperinsulinemia is potentially associated with the development of vascular sclerosis. On the other hand, the relationship between hyperinsulinemia and nephrosclerosis has not been elucidated. In this investigation clinicopathological studies were performed in 40 patients with nephrosclerosis, with special attention to the relationship between hyperinsulinemia and glomerular hypertrophy. Forty patients with biopsy-proven nephrosclerosis were divided into two groups by the 75-g oral glucose tolerance test (OGTT): group A, 2-hr plasma glucose concentration > 140 mg/dL (n = 25); group B, 140 < or = 2-hr plasma glucose < 200 mg/dL (n = 15). Patients with diabetes mellitus or diabetic nephropathy were not included. Morphometric analysis of the glomeruli revealed a significantly larger mean glomerular volume in subjects with nephrosclerosis in both subgroups. In addition, the mean glomerular volume was significantly correlated with the fasting insulin level, while no significant correlation was observed between the mean glomerular volume and creatinine clearance or degree of global sclerosis. These results indicate that hyperinsulinemia may be intimately related to glomerular hypertrophy in patients with nephrosclerosis. PMID- 8723366 TI - Influence of length of time on dialysis before grafting on kidney transplant results. AB - The outcome of kidney transplantation was evaluated in 246 nondiabetic, CsA treated recipients of primary cadaver transplant, divided into 4 groups according to length of time on dialysis: group < or = 2, 0-24 months; group 2-5, 25-60 months; group 5-15, 61-180 months; group > 15, over 180 months. The 4 groups did not differ in graft survival, proteinuria (g/die), or estimated GFR values at 1, 2, 3, 4, and 5 years after grafting. They did not differ in the frequency of cataract, hip osteonecrosis, tumors, or posttransplant diabetes mellitus at 3 years after grafting. Ocular hypertone (p < 0.02), tendon ruptures (p < 0.001), arterial occlusive disease of lower limbs (p < 0.01), cholelithiasis (p < 0.05), and chronic hepatitis--which occurred only in anti-HCV and/or HBs Ag-positive patients--(p < 0.001), were more frequent in group > 15, and in all these cases but ocular hypertone a linear trend of increasing frequencies with increasing dialytic age was statistically significant. Group 5-15 had the lowest patient survival (p < 0.02). Moreover, a progressive decline of patient survival with increasing dialytic age was noted in groups < or = 2, 2-5, and 5-15. Unexpectedly, group > 15 had remarkably good survival, and this finding denies the hypothesis of a purely linear decline of patient survival after transplantation with increasing dialytic age. PMID- 8723367 TI - Acute renal failure induced by angiotensin converting enzyme inhibitor in a patient with polyarteritis nodosa. AB - We report a patient who presented with malignant hypertension and renal failure. He was treated with lisinopril, spironolactone, and nifedipine retard for blood pressure control. Subsequent renal function showed further deterioration, but it then improved after withdrawal of the angiotensin converting enzyme inhibitor (ACE I). The diagnosis of classical polyarteritis nodosa was established with aneurysmal dilatation demonstrable in the renal vasculature. His renal impairment improved further following immunosuppressive therapy and the disease has remained inactive 4 years after first presentation. This is the first reported case of acute renal failure associated with the use of ACE I in polyarteritis nodosa. PMID- 8723368 TI - Hemodiafiltration in pentobarbital poisoning. AB - We report the use of continuous venovenous hemodiafiltration (CVVHD) in a case of massive intentional ingestion of pentobarbital associated with severe coma and hypotension. Despite a poor hemodynamic state, toxic epuration (15% of drug ingested) was obtained. However, death occurred 7 hr after the end of treatment. This report suggests that CVVHD can be effective in the treatment of massive pentobarbital intoxication. PMID- 8723369 TI - Induction of sleep. PMID- 8723370 TI - Epidemiological study on insomnia in the general population. AB - This study was conducted with a representative sample of the French population of 5,622 subjects of 15 years old or more. The telephone interviews were performed with EVAL, an expert system specialized for the evaluation of sleep disorders. From this sample, 20.1% of persons said that they were unsatisfied with their sleep or taking medication for sleeping difficulties or anxiety with sleeping difficulties (UQS). A low family income, being a woman, being over 65 years of age, being retired and being separated, divorced or widowed are significantly associated with the presence of UQS. A sleep onset period over 15 minutes, a short night's sleep and regular nighttime awakenings are also associated with UQS. Medical consultations during the past 6 months and physical illnesses are more frequent among the UQS group. The consumption of sleep-enhancing medication and medication to reduce anxiety is important: in the past, 16% of subjects had taken a sleep-enhancing medication and 16.2% a medication to reduce anxiety. At the time of the survey 9.9% of the population were using sleep-enhancing medication and 6.7% were using medication for anxiety. For most, hypnotic consumption was long-term: 81.6% had been using it for more than 6 months. PMID- 8723371 TI - Adult psychophysiologic insomnia and positive history of childhood insomnia. AB - Sixty-five adults diagnosed with "psychophysiologic chronic insomnia" following the criteria of the Association of Sleep Disorders Centers were investigated. They were subdivided into two groups based on whether onset of the insomnia had occurred in childhood or adulthood. Fifty similar-aged patients with obstructive sleep apnea syndrome (OSAS) were also investigated and served as a contrast group. All subjects were given polygraphic recordings and structured interviews, and all completed sleep questionnaires after reviewing the questions with an investigator. On many indices evaluating subjective daytime alertness and well being, the psychophysiologic insomnia patients had scores similar to the OSAS patients. There were no significant differences between the childhood-onset and adult-onset psychophysiologic insomnia patients on most of the investigated items. However, the childhood-onset psychophysiologic insomniinacs had moderately but significantly higher reports of nightmares. This group also reported having had longer sleep latencies, significantly more "fear of the dark" and more frequent nightmares during childhood than the adult-onset group. These statistically significant findings reflected only a moderate increase in subjective scores, however, and in general the childhood-onset and adult-onset psychophysiologic insomnia patients were very similar. PMID- 8723372 TI - Insomnia in the child and adolescent. PMID- 8723373 TI - Is sleep apnea syndrome in the elderly a specific entity? AB - The prevalence of apneas is increased in the elderly. In the present paper, we briefly discuss the data on epidemiology, clinical significance, morbidity and mortality of sleep apnea in the elderly. We further try to identify possible mechanisms linking aging and sleep apnea that may explain the high prevalence found in aged patients. Structural changes in upper airway, increase in pharyngeal resistance, oscillations in upper airway resistance and sleep instability are among the main factors that may be involved. Last, we present data on 389 apneics consecutively diagnosed in our sleep laboratory, separated according to whether they were less or more than 65 years of age. In the group of elderly patients that cannot be extrapolated to a random sample of the general population, sleep apnea does not seem to be different in terms of severity of breathing disorders, sleep data and therapeutic implications. Further studies are needed to determine whether or not the associated morbidity is different in the elderly; these studies could lead to different diagnosis strategies and therapeutic approaches. PMID- 8723374 TI - Interaction of serotonergic excitatory drive to hypoglossal motoneurons with carbachol-induced, REM sleep-like atonia. AB - The facilitatory effect of serotonin (5HT) on hypoglossal (XII) motoneurons is likely to be reduced during rapid eye movement (REM) sleep, when the activity of the brainstem serotonergic system reaches its nadir. Therefore, we assessed the hypothesis that application of exogenous 5HT will attenuate the REM sleep-like suppression of XII motoneurons produced in decerebrate cats by pontine microinjections of a cholinergic agonist, carbachol. Microinjections of 5HT or 5 carboxamidotryptamine into the XII nucleus increased XII nerve activity to 182 +/ 53% (standard deviation; SD) of control. Subsequent pontine microinjections of carbachol reduced XII nerve activity by 55 +/- 21% of the pre-5HT level (n = 12). Microinjections of methysergide (a 5HT antagonist) into the XII nucleus reduced XII nerve activity to 54 +/- 17% of the pre-methysergide control (n = 6). Pontine carbachol injections after methysergide further reduced XII nerve activity by 49 +/- 20% of the pre-methysergide level. Treatments with both agonists and the antagonist attenuated the carbachol-induced decrease when compared to two previous studies using the same model: 1) In experiments with no injections of serotonergic agents, pontine carbachol injections decreased XII nerve activity by 90 +/- 6% of control. 2) After enhancement of XII nerve activity by inhibitory amino acid antagonists (to 135 +/- 60%), the subsequent carbachol-induced decrease was even larger, 112 +/- 62% of control. We propose that serotonergic excitation can significantly attenuate the REM sleep-like suppression of XII nerve activity, and that this is achieved, in part, by substituting for the decreased endogenous 5HT in the XII nucleus. The study also demonstrates that other, non-serotonergic, mechanisms also contribute to the carbachol-induced suppression. PMID- 8723375 TI - Persistence of repetitive EEG arousals (K-alpha complexes) in RLS patients treated with L-DOPA. AB - Compared to normal control subjects, patients with primary restless legs syndrome (RLS) have an increased number of K-complexes, followed by bursts of electroencephalographic (EEG) alpha activity-together these are referred to as K alpha complexes. In the present study, a large proportion of K-alpha complexes was found to be temporally associated with periodic leg movements in sleep. Because many K-alpha complexes precede the onset of movements, they cannot be considered secondary to the movements themselves. The persistence of K-alpha complexes after suppression of leg movements by means of L-DOPA further supports the hypothesis that these EEG events are a primary phenomenon. The results may have clinical implications because a large proportion of RLS patients complain of non-restorative sleep even after treatment with L-DOPA. PMID- 8723376 TI - Periodic K-alpha sleep EEG activity and periodic limb movements during sleep: comparisons of clinical features and sleep parameters. AB - The K-alpha sleep electroencephalographic (EEG) phenomenon is characterized by periodic (approximately 20-40 seconds) K-complexes, immediately followed by alpha EEG activity (7.5-11 Hz) of 0.5- to 5.0-second duration. A group of 14 subjects with the periodic K-alpha anomaly was found to have a similar distribution pattern of interevent intervals as compared with previously published data for sleep-related periodic limb movements during sleep (PLMS). Sleep parameters and somatic symptoms of 30 patients with K-alpha were compared with 30 patients with PLMS. The periodic K-alpha group was predominantly female, younger, exhibiting more slow-wave sleep, gastrointestinal symptoms and muscular complaints and fewer movement arousals on overnight polysomnography. The K-alpha group presented uniformly with complaints of unrefreshing sleep, often associated with fibromyalgia and chronic fatigue syndrome. The PLMS group was predominantly male, showed greater sleep disruption and presented with a variety of sleep-related symptoms. PMID- 8723377 TI - Augmentation of the restless legs syndrome with carbidopa/levodopa. AB - Dopaminergic agents and carbidopa/levodopa have become the preferred treatment for both the restless legs (RL) syndrome and for periodic limb movements in sleep (PLMS). For once-nightly treatments with carbidopa/ levodopa, a problem with morning end-of-dose rebound increases in leg movements has been reported to occur in the about one-fourth of the patients. In our clinical studies a previously unreported but far more significant problem of markedly augmented RL symptoms occurred in the afternoon and the evening prior to taking the next nightly dose. A systematic prospective evaluation of this augmentation in 46 consecutive patients treated with carbidopa/ levodopa for RL syndrome or PLMS disorder found this augmentation to be the major adverse effect of treatment. Augmentation occurred for 31% of PLMS patients and 82% of all RL patients. It was greater for subjects with more severe RL symptoms and for patients on higher doses (> or = 50/200 mg carbidopa/levodopa) but was unrelated to gender, age or baseline severity of PLMS. This augmentation was severe enough to require medication change for 50% of the RL patients and 13% of PLMS patients. Augmentation resolved with cessation of the medication and could be minimized by keeping the dose low. PMID- 8723379 TI - Are periodic leg movements associated with clinical sleep disturbance? AB - We examined 67 patients with periodic leg movement (PLM) disorder who were seen in a university-based sleep center. The most common reasons for coming to the sleep center were insomnia, sleepiness and a request for an evaluation for possible sleep apnea. There was a significant positive correlation between PLM arousal index and age but no association with gender. Approximately one-quarter of patients were under age 30. The multiple sleep latency test (MSLT) revealed borderline normal wakefulness in the group as a whole (sleep latency of 10.2 +/- 0.9 minutes), and there was no significant correlation between the PLM arousal index and either the MSLT mean sleep latency or a measure of subjective sleepiness. Similarly, the PLM arousal index did not differentiate those who entered with chief complaints of insomnia or sleepiness. There was no significant difference in the PLM arousal index in those who reported that they did or did not awaken refreshed in the morning. In summary, in this clinical population we found no significant association between the PLM arousal index and the subjective complaint of disturbed sleep, an objective measure of daytime sleepiness or a sense of awakening refreshed in the morning. Other interesting observations included the relatively high frequency of a PLM index > 5 in patients under 30 years old and a relatively high rate of past treatment for depression. PMID- 8723378 TI - L-DOPA/carbidopa for nocturnal movement disorders in uremia. AB - Restless legs syndrome (RLS) and periodic limb movements during sleep (PLMS) are sleep disorders that are common and distressing to uremic patients. There are few data regarding effective treatment in this population. Five chronic hemodialysis patients completed a double-blind, placebo-controlled, crossover study using a single bedtime dose of controlled release L-DOPA/carbidopa (100/25 mg) for treatment of RLS and sleep disruption. Leg movements per hour of sleep and percentage of sleep time accompanied by leg movements were decreased with treatment (101.0 +/- 29.1 events/hour on placebo vs. 61.0 +/- 28.3 events per hour on drug, p = 0.006; and 15.1 +/- 4.9% of sleep time with leg movements on placebo vs. 8.6 +/- 4.0% on drug, p = 0.014). In addition, arousals associated with leg movements (mean 209 +/- 49 events on placebo, mean 108 +/- 46 events on drug) and the leg movement arousal index (mean 59 +/- 23 events/hour on placebo, mean 23 +/- 9 events/hour on drug) were decreased by active medication (p = 0.03 and 0.04, respectively). Patients, however, continued to have very disrupted sleep and we could not document consistent subjective or objective improvement in overall sleep except for an increase in slow-wave sleep (SWS) from 9.0% to 22.8% (p = 0.01). The patterns of movements during sleep were not uniform in different patients, and the movements, although often periodic, were much longer than defined for PLMS. Because of this, finding suitable objective parameters to analyze was problematic. Measuring the percentage of sleep time during which there were leg movements was probably the most efficient and reproducible means of quantitating this disorder. Thus, although controlled-release L-DOPA/carbidopa at a dose of 100/25 mg given once nightly reduced leg movements and increased SWS, sleep continued to be disrupted. Whether higher doses or more frequent dosing is effective requires further investigation. PMID- 8723380 TI - Management of restless legs syndrome with gabapentin (Neurontin) PMID- 8723381 TI - Chronic hiccups and sleep. AB - To explore the effect of sleep on hiccups, we studied eight patients aged 20-81 years, all males with chronic hiccups lasting 7 days to 7 years, by means of overnight polysomnography. The incidence of new bouts of hiccups and the likelihood of hiccups being present were both highest in wakefulness and became progressively lower through stages I-IV of slow wave sleep (SWS) to rapid eye movement sleep (REMS). There was a significant tendency for hiccups to disappear at sleep onset and REMS onset. Of all 21 bouts of hiccups that were observed to stop, 10/21 did so during an apnea or hypopnea. Frequency of hiccups within a bout slowed progressively from wakefulness through the stages of SWS to REMS. For the whole group, mean frequency decreased significantly from wakefulness [(25.6 +/- 12.1), (mean +/- SD)] to sleep onset or stage I (22.3 +/- 12.2). Sleep latency was increased from 8 +/- 16.3 minutes when hiccups were absent to 16.35 +/- 19.9 minutes when it was present. Sleep efficiency was poor because of long waking periods, and there were deficiencies of both SWS and REMS. Hiccups themselves were not responsible for any arousals or awakenings. We conclude that neural mechanisms responsible for hiccups are strongly influenced by sleep state and that hiccups disrupt sleep onset but not established sleep. PMID- 8723382 TI - Language in dreaming and regional EEG alpha power. AB - In an intensive single-subject design, electroencephalographic (EEG) alpha power and receptive and expressive language in dreaming were studied in 12 dreams during rapid eye movement (REM) sleep on 12 separate nights. Bilateral EEG was recorded continuously from 21 sites and digitized. We used the Fast Fourier transformation (FFT) for power spectral analysis to measure EEG power in the alpha frequency range (8-12 Hz) at each of the EEG sites. The subject was awakened after about 14 minutes into the second REM period, and dream reports were collected. We scored the dream reports for expressive and receptive language. The lower the alpha power on the left sides of those homologous pairs that roughly correspond to Broca's (C3) or Wernicke's area (P3), the more expressive or receptive language in dream reports. The largest difference between the correlation of the left and that of the right homologous pair of regions was found in the central (C3, C4) area for expressive language and in the parietal (P3, P4) area for receptive language. Our finding suggests lateralized and localized cortical activation in relation to language in dreaming. PMID- 8723383 TI - Sleep-induced breathing instability. University of Wisconsin-Madison Sleep and respiration Research Group. AB - We present a view of the neuromechanical regulation of breathing and causes of breathing instability during sleep. First, we would expect transient increases in upper airway resistance to be a major cause of transient hypopnea. This occurs in sleep because a hypotonic upper airway is more susceptible to narrowing and because the immediate excitatory increase in respiratory motor output in response to increased loads is absent in non-REM sleep. Secondly, sleep predisposes to an increased occurrence of ventilatory "overshoots", in part because abruptly changing sleep states cause transient changes in upper airway resistance and in the gain of the respiratory controller. Following these ventilatory overshoots, breathing stability will be maintained if excitatory short-term potentiation is the prevailing influence. On the other hand, apnea and hypopnea will occur if inhibitory mechanisms dominate following the ventilatory overshoot. These inhibitory mechanisms include: a) hypocapnia-if transient, will inhibit carotid chemoreceptors and cause hypopnea, but if prolonged will inhibit medullary chemoreceptors and cause apnea; b) a persistent inhibitory effect from lung stretch; c) baroreceptor stimulation, from a transient rise in systemic blood pressure immediately following termination of apnea or hypopnea may partially suppress the accompanying hyperpnea; d) depression of central respiratory motor output via prolonged brain hypoxia. Once apneas are initiated, reinitiation of inspiration is delayed even though excitatory stimuli have risen well above their apneic thresholds, and these prolonged apneas are commonly accompanied by tonic EMG activation of expiratory muscles of the chest wall and upper airway. PMID- 8723384 TI - EEG frequency changes during sleep apneas. AB - To study the effect of transient, apnea-induced hypoxemia on electrocortical activity, five patients with severe obstructive sleep apnea syndrome (OSAS) were investigated during nocturnal sleep. Polysomnographic and simultaneous digitized electro encephalographic (EEG) recordings for topographic and compressed spectral array analysis were made. The EEG recordings were timed exactly to respiratory events. During nonrapid eye movement (NREM) apnea, delta band amplitude increased, starting on average 13 seconds after the apnea onset. Average differences were 268% between initial and maximal values and 202% between initial and final values. In contrast, significant increases in delta amplitudes between the onset and end of REM apneas did not occur, although some caused deep oxygen desaturations. Changes in delta activity were not correlated to NREM apnea duration or degree of desaturation. These results indicate that the increased delta activity during NREM apneas may not be caused by arterial hypoxemia. It could instead be due to either an arousal mechanism, since arousals may be preceded by slow waves in EEG, or to a breakthrough of slow-wave-sleep activity. The sleep disturbance in severe OSAS may create such a propensity for slow-wave sleep that stages pass much more rapidly than in normal persons. PMID- 8723385 TI - Cardiogenic oscillations and airway patency. PMID- 8723386 TI - Bibliography of recent literature in sleep research. PMID- 8723387 TI - Gene transcription through activation of G-protein-coupled chemoattractant receptors. AB - Receptors for leukocyte chemoattractants, including chemokines, are traditionally considered to be responsible for the activation of special leukocyte functions such as chemotaxis, degranulation, and the release of superoxide anions. Recently, these G-protein-coupled serpentine receptors have been found to transduce signals leading to gene transcription and translation in leukocytes. Transcription factors, such as NF kappa B and AP-1, are activated upon stimulation of the cells with several chemoattractants at physiologically relevant concentrations. Activation of transcription factors through these receptors involves G-protein coupling and the activation of protein kinases. The underlying signaling pathways appear to be different from those utilized by TNF alpha, a better characterized cytokine that induces the transcription of immediate-early genes. Chemoattractants stimulate the expression of several inflammatory cytokines and chemokines, which in turn may activate their respective receptors and initiate an autocrine regulatory mechanism for persistent cytokine and chemokine gene expression. PMID- 8723388 TI - Sequential steps in Tat trans-activation of HIV-1 mediated through cellular DNA, RNA, and protein binding factors. AB - The regulation of HIV expression is controlled by the activity of the Long Terminal Repeat (LTR). Trans-activation by the virally encoded Tat protein is one of the main mechanisms of LTR activation. Tat binds to its target, TAR RNA, and cellular proteins that bind the LTR, Tat, or TAR RNA are important components of the trans-activation process. We will review the factors that have been characterized for a possible involvement in this mechanism. Whereas LTR binding proteins consist of Sp1 and TBP, a large number of factors that bind TAR RNA have been isolated. We have previously cloned two of them by RNA probe recognition: TRBP and La. We have shown that the in vitro and in vivo binding of TRBP to TAR RNA correlates with a constant expression of the protein during HIV-1 infection. Several proteins that interact with Tat have mainly positive, but some negative, effects on trans-activation. Genetic studies have defined that human chromosome 12 encodes a protein that will allow trans-activation in rodent cells. The binding and the functional data about these proteins suggest sequential steps for the Tat trans-activation mechanism. Each of these intracellular molecular events could be the target for molecular intervention against the virus. PMID- 8723389 TI - From repression domains to designer zinc finger proteins: a novel strategy of intracellular immunization against HIV. AB - Tissue-specific gene regulation of eukaryotic organisms is to a large extent mediated by transcription factors that interact with genomic DNA sequences in a sequence-specific manner. The purpose of this synopsis is to put forward the potential of designer zinc finger proteins in treating infections of human immunodeficiency virus (HIV). Artificial transcription factors containing designer zinc finger structures fused to activator or repressor domains have been designated Transcription Response Modifiers (TRMs). The principle of engineering TRMs has been derived from the analysis of human Kruppel-type zinc finger genes and their products. Our research efforts encompass two fascinating features that are displayed by the human Kruppel-type zinc finger protein KOX1: 1) the Kruppel type zinc finger domains display rules of sequence-specific DNA recognition, and 2) the evolutionarily conserved Kruppel-associated box (KRAB) presents one of the strongest transcriptional repressors identified so far in mammalian organisms. The KRAB repressor activity is postulated to be mediated through co-repressor molecules, such as Silencing Mediating Protein-1 (SMP-1). Thus, the structural organization and functional analysis of zinc finger proteins revealed principles of zinc finger transcription factors that are applicable for reducing the viral load in individuals infected with HIV. In this article, a novel concept of generating therapeutic proteins is outlined that might be conceptually promising in modulating gene expressions of any kind. PMID- 8723390 TI - The expression of a small fraction of cellular genes is changed in response to histone hyperacetylation. AB - Posttranslational modifications of histones in chromatin are emerging as an important mechanism in the regulation of gene expression. Changes in histone acetylation levels occur during many nuclear processes such as replication, transcriptional silencing, and activation. Histone acetylation levels represent the result of a dynamic equilibrium between competing histone deacetylase(s) and histone acetylase(s). We have used two new specific inhibitors of histone deacetylase, trichostatin A (TSA) and trapoxin (TPX), to probe the effect of histone hyperacetylation on gene expression. We confirm that both drugs block histone deacetylase activity and have no detectable effects on histone acetylation rates in human lymphoid cell lines. Treatment with either TSA or TPX results in the transcriptional activation of HIV-1 gene expression in latently infected cell lines. In contrast, TSA and TPX cause a rapid decrease in c-myc gene expression and no change in the expression of the gene for glyceraldehyde-3 phosphate dehydrogenase (GAPDH). Using differential display to compare the differences in gene expression between untreated cells and cells treated with TSA, we found that the expression of approximately 2% of cellular genes (8 genes out of approximately 340 examined) changes in response to TSA treatment. These results demonstrate that the transcriptional regulation of a restricted set of cellular genes is uniquely sensitive to the degree of histone acetylation in chromatin. PMID- 8723392 TI - Neurohumoral mechanisms in heart failure: a central role for the renin angiotensin system. AB - Congestive heart failure (CHF) is often the result of a progressive deterioration in cardiac function associated with marked alterations in peripheral physiology. The neurohumoral mechanisms involved in these alterations have the paradoxical role of serving simultaneously as adaptive, compensatory changes and as major contributing elements in the progression of CHF. This review discusses some of the clinical and experimental data that describe the time course of these neurohumoral mechanisms, with an emphasis on the renin-angiotensin system (RAS). The central role of the RAS in the pathophysiology of heart failure is discussed in relation to its interaction with other neurohumoral systems, its role in the peripheral vascular alterations, and its role in the myocardial alterations in CHF. In addition, these effects are discussed in the context of both the circulating and local RAS and the ability of pharmacologic interventions that modulate the RAS to modify the course of CHF favorably, especially the angiotensin-converting enzyme (ACE) inhibitors. An understanding of the intricate interactions of these neurohumoral mechanisms in CHF has already led to the development of therapies that reduce morbidity and mortality and provides the basis for continuing advances in the treatment of CHF. PMID- 8723393 TI - Importance of endothelial function in chronic heart failure. AB - Chronic heart failure (CHF) is associated with neurohumoral activation and alterations of the peripheral circulation. Several mechanisms are involved in the impaired peripheral perfusion, including increased sympathetic tone. Recent data suggest an important role of the endothelium for peripheral perfusion in CHF. Endothelium-dependent dilation of resistance vessels is blunted in patients with severe CHF and may be involved in the impaired reactive hyperemia in these patients. In conductance vessels, flow-dependent dilation is reduced in CHF, reflecting endothelial dysfunction of large conduit vessels. To investigate endothelial function in humans in vivo, agents such as acetylcholine are used to stimulate the release of endothelium-derived nitric oxide (NO). Conversely, N mono-methyl-L-arginine (L-NMMA), a specific inhibitor of NO synthesis from L arginine, decreases forearm blood flow by inhibiting the basal release of NO. Conversely to the impaired stimulated release of NO by acetylcholine, the decrease in blood flow induced by L-NMMA appears to be exaggerated in CHF. The blood flow response to nitroglycerin or sodium nitroprusside, both endothelium independent vasodilators, is usually preserved in patients with nonedematous CHF, indicating a normal response of the vascular smooth muscle of resistance vessels to exogenous NO. Therefore, impaired endothelium-dependent dilation of peripheral resistance vessels emerges in CHF, suggesting a reduced release of NO on stimulation. Endothelial dysfunction may therefore be involved in the impaired vasodilator capacity in the peripheral circulation, e.g., during exercise. In contrast, the basal release of NO from endothelium of resistance vessels appears to be enhanced and may play an important compensatory role in CHF. PMID- 8723391 TI - Human nuclear receptor heterodimers: opportunities for detecting targets of transcriptional regulation using yeast. AB - Nuclear receptors are model transcription factors. This highly conserved superfamily of ligand binding transcription factors includes estrogen, progesterone, retinoic acid, thyroid hormone, vitamin D receptors, and several orphan receptors. Nuclear receptors function as homodimers, heterodimers, or monomers. Human thyroid hormone, retinoic acid, vitamin D, and several orphan receptors prefer to work as heterodimers with retinoic X receptor (RXR). RXR function is regulated by its cognate ligand 9-cisretinoic acid. In some cases heterodimers of RXR are subject to regulation by two different ligands. Mammalian cells are not entirely suited to study pure heterodimeric functions because they contain a repertoire of endogenous receptors and their ligands. Yeast does not contain nuclear receptors or its ligands. Ligand-dependent function of several human nuclear receptors has been reconstructed in yeast. Yeast can be used as a model system to dissect interaction between various heterodimeric partners. The molecular genetics and the speed of doing the experiments in yeast allows us to rapidly clone mammalian cofactors that prefer to work with different heterodimeric partners. Once the human genome sequence is complete, we predict that the total number of human nuclear receptors will increase from 150 to 500. Novel and efficient cell-based systems will be needed to understand the function of orphan receptors. Yeast is an ideal system to identify pure heterodimeric partners and discover novel ligands for orphan receptors. The advantages and disadvantages of yeast and mammalian system to study nuclear receptor function are discussed. PMID- 8723394 TI - Correction of remodeling and function of small arteries in human hypertension by cilazapril, an angiotensin I-converting enzyme inhibitor. AB - Angiotensin II may contribute to the altered structure and function of small arteries. We proposed that angiotensin I-converting enzyme (ACE) inhibitor treatment could induce a regression of vascular remodeling. A double-blind trial was performed comparing effects of the ACE inhibitor cilazapril with the beta blocker atenolol on small arteries obtained from biopsy specimens of subcutaneous gluteal fat. Nine patients with essential hypertension were randomized to cilazapril and eight to atenolol. Blood pressure was below 140/95 mm Hg under treatment for the duration of the study in all patients. Media-to-lumen ratio of small arteries of the patients, which before treatment was significantly higher than in normotensive subjects, was corrected after 1 year of treatment in the cilazapril group. There was no change in the increased media-to-lumen ratio of small arteries in the atenolol group, even after 2 years of treatment. Attenuated constrictor responses to endothelin-1 returned to normal only in the patients treated with cilazapril. Endothelium-dependent relaxation responses to acetylcholine were slightly depressed in hypertensive patients and improved in the cilazapril-treated group, but remained blunted in the arteries of the atenolol-treated patients. Treatment with cilazapril corrects small artery remodeling and endothelium-related functional abnormalities of gluteal subcutaneous small arteries in hypertensive patients. It remains to be demonstrated whether these apparently beneficial effects translate into reduced morbidity and mortality in hypertension. PMID- 8723395 TI - Clinical benefit of angiotensin-converting enzyme inhibitors in chronic heart failure. AB - The ideal therapy for patients with chronic heart failure should reduce symptoms related to pulmonary congestion or low perfusion, prevent the progression of left ventricular dysfunction and, ultimately, should reduce mortality. Extensive studies in humans have investigated the effects of angiotensin-converting enzyme (ACE) inhibitors on these goals of therapy. As an example, the ACE inhibitor cilazapril significantly improved exercise tolerance, as borne out by a meta analysis of six placebo-controlled, randomized 3-month trials. Comparison of the effects of cilazapril and captopril vs. placebo in one of the trials documented similar improvement in exercise tolerance (14 vs. 17%). Results from other randomized comparative trials suggest that the improvement in symptoms represents a class effect of ACE inhibitors. A beneficial effect of ACE inhibition on the progression of left ventricular dysfunction has also been demonstrated in the SOLVD trial, and a reduction of mortality has been amply documented in several mortality trials (CONSENSUS I, SOLVD, V-HeFT-II, SAVE, AIRE, SMILE) in patients with or without preceding myocardial infarction. Reports that ACE inhibitors also reduce the incidence of reinfarction after myocardial infarction have not been confirmed in all studies but raise the interesting concept that ACE inhibition may interact, in a beneficial but thus far not well-understood way, with key processes in the development of atherosclerosis, thereby preventing plaque rupture, thrombus formation, and myocardial infarction. Taken together, a large database convincingly demonstrates that ACE inhibitors are effective not only in improving symptoms but also in the prevention of progression of left ventricular dysfunction, in the reduction of mortality, and possibly in stabilizing the atherosclerotic disease process. PMID- 8723396 TI - Improvement of vagal tone by ACE inhibition: a mechanism of cardioprotection in patients with mild-to-moderate heart failure. AB - A total of 35 patients in sinus rhythm and with mild-to-moderate congestive heart failure (CHF) (NYHA II-III) aged 53 (+/- 3) years were examined before therapy with angiotensin-converting enzyme (ACE) inhibitors. Of these patients, 16 were reexamined after therapy with ACE inhibitors for 17 +/- 3 days. The relation of hemodynamic alterations to vagal tone was assessed and the influence of parasympathetic (baroreflex activation) tone on survival was evaluated. Only hemodynamic responders to ACE inhibition showed a significant increase of vagal tone from 1.4 +/- 0.4 to 3.6 +/- 1.2 ms/mm Hg (p < 0.01). The magnitude of increase of vagal tone was dependent on the baseline level. All 35 patients were discharged on ACE inhibitors and were followed for 56 months or longer. We compared patients whose hearts survived (20 patients) with those whose hearts did not (15 patients). Twelve patients died and three underwent cardiac transplantation. The two groups differed (p < 0.05) in terms of mean arterial blood pressure (98 +/- 3 vs. 90 +/- 3 mm Hg), heart rate (82 +/- 2 vs. 93 +/- 4 beats/min), and mean pulmonary artery pressure (24 +/- 3 vs. 35 +/- 2 mm Hg). Cardiac index, stroke volume index, and right atrial pressures were not different. Heart survivors had lower (p < 0.05) renin values (3.6 +/- 0.8 vs. 9.0 +/- 3.6 Ang-I/ml/h) but norepinephrine values at baseline were not different. Baroreflex sensitivity was lower (p < 0.02) in survivors than in nonsurvivors (1.3 +/- 0.2 vs. 2.3 +/- 0.3 ms/mm Hg). We then calculated the mortality risk in relation to baroreflex sensitivity at the median BS of 1.6 ms/mm Hg. Survival was different (p < 0.004) between the resulting two groups: 2 of 15 subjects (13%) with high baroreflex sensitivity died, and 13 of 20 patients (65%) whose baroreflex sensitivity was less than 1.6 ms/mm Hg died. When systemic blood pressure, pulmonary artery pressure, stroke volume index, plasma norepinephrine concentrations, and baroreflex sensitivity were entered into a Cox proportional hazards regression, only systolic blood pressure and plasma norepinephrine values predicted survival (p < 0.001). We concluded that high vagal tone is correlated with a good prognosis in patients with CHF. ACE inhibitor therapy can increase the vagal tone significantly. This may alter the incidence of sudden cardiac death and thereby improve prognosis. PMID- 8723397 TI - Quality of life with ACE inhibitors in chronic heart failure. AB - The randomized trials assessing the effect of angiotensin-converting enzyme (ACE) inhibitors in chronic heart failure (CHF) are reviewed. The Minnesota Living with Heart Failure Questionnaire has demonstrated the benefits of enalapril in some but not all circumstances and the Yale Dyspnea-Fatigue Index improves with lisinopril. A recent trial of both cilazapril and captopril vs. placebo employed the Sickness Impact Profile and supports the concept that ACE inhibitors have a small (and in this trial nonsignificant) beneficial effect on mobility. Other vasodilators and inotropes may also have small benefits on quality of life, such that comparisons of an ACE inhibitor with vasodilators, as was done in the V-HeFT II trial, fail to reveal any different effects on quality of life. PMID- 8723398 TI - Cardiovascular hypertrophy: role of angiotensin II and bradykinin. AB - Angiotensin II can raise blood pressure rapidly by inducing direct vasoconstriction and by activating the sympathetic nervous system via central and peripheral mechanisms. In addition, this peptide may act as a growth factor to cause vascular and cardiac hypertrophy (CVH). The structural changes caused by hypertension can therefore be amplified by angiotensin II. Blockade of angiotensin II generation with angiotensin-converting enzyme (ACE) inhibitors appears to be particularly effective in preventing the development of cardiovascular hypertrophy. This beneficial effect might be related to some extent to local accumulation of bradykinin. ACE is one of the enzymes physiologically involved in bradykinin degradation. Treatment of hypertensive rats with a selective bradykinin antagonist can attenuate the blood pressure lowering effect of ACE inhibition and render less effective the prevention of intimal thickening after endothelial removal from the rat carotid artery. Bradykinin is a vasodilator that acts by increasing the release of endothelium derived factors such as nitric oxide and prostacyclin, which may have antiproliferative activity. However, blockade of the renin-angiotensin system with an angiotensin II subtype 1-receptor antagonist is also effective in preventing cardiac hypertrophy and neointimal proliferation after endothelial injury. Therefore, the exact contribution of bradykinin to the beneficial effects of ACE inhibition on cardiovascular hypertrophy remains to be further explored. PMID- 8723399 TI - Amputation versus reconstruction in traumatic defects of the leg: outcome and costs. AB - This retrospective review covers global aspects of reconstructive efforts to salvage severely injured legs. Eighteen patients with traumatic lower leg amputation were compared to 21 patients who underwent complex microvascular reconstruction. The mean number of interventions was 3.5 for amputation and 8 for reconstruction (p < 0.009). Total rehabilitation time was 12 months for amputation and 30 months for reconstruction (p < 0.009). Changes in lifestyle were consistently more important in the amputee group. The mean annual hospital costs for amputated patients were 15,112 Swiss Francs (SD 7,094 SF) for the first 4 years. The mean annual hospital costs for reconstructed patients were 17,365 Swiss Francs (SD 8,702 SF) for the first 4 years. Fifty-six percent of the amputees and 19% of the reconstructed patients were retrained to a different profession (p < 0.025). Fifty-four percent of the amputees and 16% of the reconstructed patients were drawing an extremely costly and life long invalidity pension (p < 0.02). We conclude that for potentially salvageable legs reconstruction is advisable because the functional outcome was better than for amputation and there was no permanent social disintegration due to the long treatment. Total costs (including pensions) for reconstruction were far lower than for amputation. PMID- 8723400 TI - Incidence of deep-vein thrombosis in patients with fractures of the lower extremity distal to the hip. AB - Our purpose was to determine the incidence of deep-vein thrombosis (DVT) in patients who have had early operative fixation of fractures of the lower extremity distal to the hip. There is a high incidence of distal thrombosis in patients who have undergone early operative fixation of lower-extremity fractures. The incidence of DVT is higher with proximal extremity fractures than with distal extremity fractures. Most clots are occult and do not progress clinically. Given the high incidence of DVT with femoral and tibial plateau fractures, older age, and longer operating times, anticoagulation prophylaxis may be indicated. Ours was a prospective incidence study. All patients who had had early operative fixation of lower-extremity fractures were eligible (n = 176). Seventy-four were excluded based on specific criteria. The remaining 102 patients underwent lower-extremity venography an average of 9 days after operative fixation of their fractures (range 3-22 days). Eight patients had bilateral lower extremity fractures. The patients were followed clinically for 6 weeks to identify complications related to venous disease. The overall incidence of clinically occult DVT was 28%. Of the represented fractures, 40% were of the femoral shaft (eight of 20), 43% of the tibial plateau (12 of 28), 22% of the tibial shaft (12 of 54), and 12.5% of the tibial plafond (one of eight). Four of the thrombi were proximal to the popliteal fossa. Four of the patients had clinical evidence of pulmonary embolism while in the hospital. One of them had objective confirmation on further testing. No patient had clinical evidence of pulmonary emboli as an outpatient by follow-up criteria. The incidence of DVT has not been determined for patients with early stabilization of lower-extremity fractures. This study suggests a higher DVT incidence in more proximal fractures, but little risk of embolization. Thrombus formation proximal to the popliteal fossa is rare. Older age, longer operating times, and longer times before fracture fixation all correlate with an increased incidence of DVT. PMID- 8723401 TI - Improvement of the bone-pin interface with hydroxyapatite coating: an in vivo long-term experimental study. AB - The present study was designed to comparatively evaluate the bone-pin interface in a long-term unloaded experimental study in two groups of pins, uncoated and coated with hydroxyapatite. Forty pins made of stainless steel were used. Half of the pins were plasma-sprayed with hydroxyapatite, and the other half remained uncoated. Four adult sheep were selected. Each sheep tibia was implanted with five pins. Two sheep were euthanized 4 months after surgery and the remaining two 12 months after surgery. Extraction torque was higher in the hydroxyapatite coated pins compared with the uncoated ones at both 4 (p < or = 0.0005) and 12 months (p < or = 0.0005) after implantation. The histological patterns observed in the sheep euthanized 4 and 12 months after implantation were very similar. An extensive bony coverage of the hydroxyapatite-coated pins without any coating resorption and delamination from the metallic substrate was observed. Fibrous tissue encapsulation was found in the uncoated pins. These results demonstrate that the hydroxyapatite coating significantly improved the bone-pin interface. A similar improvement of bone-pin interface rigidity in many clinical situations is likewise possible. PMID- 8723402 TI - The use of an anterior incision of the meniscus for exposure of tibial plateau fractures requiring open reduction and internal fixation. AB - The purpose of this study was to examine the use of an anterior incision of the meniscus for exposure of tibial plateau fractures. We studied 27 fractures of the proximal tibia treated with open reduction and internal fixation (ORIF). There were nine unicondylar fractures (five A-O B2; four A-O B3) fixed with plates and screws and 18 bicondylar fractures (seven A-O C1; five A-O C2; six A-O C3) fixed with combination internal and external fixation. Length of follow-up averaged 26 months. All patients were treated with an anterior incision of the meniscus and retraction with the condyle. Of the 18 bicondylar fractures, nine severely displaced fractures were found to have peripherally detached menisci. Unicondylar fractures did not display this finding. After fixation, menisci were repaired at the periphery and sewn to the original anterior insertion. The repair begins posteriorly and advances the cartilage to ensure anatomic placement. There were four medial and 23 lateral menisci in this series. Ten patients underwent knee arthroscopy 6 months to 2 years post-ORIF as a routine procedure during hardware removal. All menisci were found to be healed to the periphery and were stable. There were no gross tears. In one patient, the anterior meniscal incision could be visualized. No patients developed mechanical symptoms either in postoperative rehabilitation or postoperative follow-up at a maximum of 6 years. All patients had > 125 degrees of motion. Less motion when compared with the normal knee was felt to be related to more complex fracture patterns. In conclusion, the anterior meniscal incision allows for excellent exposure of severe proximal tibia fractures. This technique allows for anatomic meniscal repair and early rehabilitation. Arthroscopic examination confirms peripheral meniscal healing. No patient experienced clinical symptoms of meniscal pathology. PMID- 8723403 TI - Relationship of mechanical factors to the strength of proximal femur fractures fixed with cancellous screws. AB - The decision of whether to attempt screw fixation of a femoral neck fracture is based partly on the estimated strength of the fixed bone/implant construct in relation to the loads it will be required to bear. The goal of this study was to determine in vitro the relation of the following biomechanical factors to the strength of internally fixed femoral neck fractures subjected to cyclic and failure loading: (a) square of the density of cancellous bone in the femoral head, (b) percent comminution of the inferior fracture surface, (c) moment arm of the joint force, or distance from the axis of the joint force to the fracture surface, and (d) orientation angle of the fracture surface in the medial/lateral plane relative to the axis of the femoral shaft. Femoral neck fractures were created in each of 38 fresh cadaveric proximal femora using a dropweight or with a materials testing machine. After sustaining a displaced fracture, fixation was achieved using three cannulated cancellous bone screws. The fixed femur was then subjected to 10,000 cycles of a sinusoidially varying load acting on the femoral head, parallel to the femoral shaft, with an initial peak magnitude of 2.2 times body weight, while the hip was flexed, extended, and rotated to mimic some motions of gait. Muscle loading was not simulated. The magnitude of the peak load decreased as the femoral head displaced during cycling. The mean of the peak load for each cycle over the duration of the test was defined as the average load. Following cycling, the bone/screw construct was loaded to failure in the same direction, and this measurement was termed the maximum load. Average and maximum load were then correlated to the four biomechanical factors using a multiple regression analysis. These factors correlated to a high degree with average force (R2 = 0.771; p < 0.0001) and to a lesser but still significant degree with maximum force (R2 = 0.458; p = 0.012), demonstrating that they can be used to estimate the strength of fixation under these loading conditions. The strongest correlation for average force was with fracture angle (p = 0.005) and for failure force was with moment arm length (p = 0.072). PMID- 8723404 TI - Prevention of heterotopic bone formation after acetabular fracture fixation by single-dose radiation therapy: a preliminary report. AB - A common complication after acetabular fracture fixation is formation of heterotopic ossification (HO) in the soft tissues about the hip. We are reporting on the use of a single, low-dose radiation therapy regimen to prevent HO in the population of postoperative acetabular fracture patients. Thirty acetabular fracture patients who were at risk for HO after surgery were reviewed with an average follow-up of 12 months. Twenty-one of these patients received radiation therapy in the postoperative period according to a single dose protocol. Nine patients did not receive any prophylactic treatment for the prevention of HO during the same period. One patient from each of these categories was lost to follow-up. Of the 20 treated and followed patients, none developed HO higher than grade 2, and nine had no HO at all. None of the treated patients had any functional deficit due to HO, and none had any complications related to radiation treatment. Of the untreated patients, all had grade 2 HO or above. Three developed grade 4 HO and have undergone reoperation for resection of HO. Study weakness include the fact that the non-treated patients were significantly different from the treated patients in several ways; including the incidence of multiple trauma, head injury, and delay to surgery. No attempt was made to randomize patients prospectively, nor to directly compare the two groups. However, we can conclude, based on comparison with reported rates of HO formation, that single, low-dose radiation therapy is a safe and effective method for prevention of HO in the postoperative acetabular fracture patient. It is as effective as previously reported multidose regimens. PMID- 8723405 TI - Anatomic considerations of plate-screw fixation of the anterior column of the acetabulum. AB - Fifteen cadaveric adult bony hemipelvis specimens and 30 adult dry bone specimens were obtained to evaluate the configuration of the anterior column of the acetabulum and to develop a safe path for screw placement into it. Each cadaveric specimen was sectioned at 1-cm intervals, beginning at the level of the inferior border of the acetabulum (junction between the anteroinferior edge of the acetabulum and the most anterolateral edge of the superior ramus of the pubic bone). The plane of the cross-section was perpendicular to the anterior column. The projection of the medial acetabular boundary on the anterior column was determined by analysis of each cross-section. Results showed that the average width of the anterior column at 1.0, 2.0, and 3.0 cm superior to the inferior acetabular boundary is 31.0 +/- 4.7, 34.2 +/- 5.1, and 39.4 +/- 6.2 mm, respectively. At 1.0 cm superior to the inferior margin of the acetabulum, the average medial angulation for 0.5-, 1.0-, and 1.5-cm entry points lateral to the pelvic brim were 24.9 +/- 4.4 degrees, 35.5 +/- 5.2 degrees, and 44.4 +/- 6.6 degrees, respectively. At 2.0 cm superior to the inferior acetabular margin, the corresponding average medial angulation for 0.5-, 1.0-, 1.5-cm entry points were determined to be 29.2 +/- 5.5 degrees, 38.6 +/- 5.9 degrees, and 48.1 +/- 5.7 degrees, respectively. At 3.0 cm superior to the inferior acetabular margin, these angles were found to be 20.7 +/- 4.3 degrees, 29.4 +/- 6.0 degrees, and 39.3 +/- 5.9 degrees, respectively. All of the above mentioned angles are with respect to the perpendicular of the longitudinal axis of the anterior column without violation of the hip joint. Screws placed 1.0 cm lateral to the pelvic brim at the levels of 1.0, 2.0, 3.0, and 4.0 cm superior to the inferior acetabular margin and directed perpendicular to the anterior column penetrated the hip joint. PMID- 8723406 TI - Gunshot fractures of the humeral shaft treated with external fixation. AB - In a prospective study, we assessed 38 consecutive gunshot fractures of the humeral shaft treated between 1 April 1990 and 30 June 1994. The average age was 34 (range, 16-60) years. Average follow-up was 31 (range, 8-50) months. Low velocity fractures occurred in 35 patients. All fractures were comminuted and displaced. Nerve injuries (radial and/or median) occurred in eight and vascular injuries in three patients. All three vascular injuries had combined nerve injuries. The arteries and nerves were explored; only one artery was found severed and required repair; two arteries and all explored nerves were in continuity. All patients had minimal debridement, antibiotic therapy and stabilization of the fracture with an external fixator. The wounds healed by granulation in 27 patients; seven had secondary closure, and four had split skin grafts. The external fixator was left in place for 6-24 (average, 16) weeks. In 34 patients union occurred between 12 and 24 (average, 16) weeks. Two patients with delayed union required bone grafting. Nonunion occurred in two patients (5%). Full recovery of the nerve palsy without further intervention was observed in seven of the eight cases. Superficial pin track infection was present in five patients; two had deep wound sepsis, and one had bone sepsis. We recommend this treatment for low-velocity humeral shaft fractures. PMID- 8723407 TI - Effects of periosteal stripping on healing of segmental fractures in rats. AB - The present study was undertaken to assess the effect of periosteal detachment upon the healing of segmental diaphyseal fractures. In rats we produced two standardized partial osteotomies with an 8-mm intermediary fragment in the femoral diaphysis. The osteotomies were then manually broken, retaining the periosteal and muscular attachment on the medial side in one group. In the other group, the segment was stripped of periosteum circumferentially. The fractures were stabilized with 1.6-mm steel pins, and the rats were allowed free movement. After 4, 8, and 12 weeks eight rats in each group were sacrificed, and callus formation, mechanical parameters, and bone blood flows were evaluated. The area of callus in the fractures with periosteal attachment was significantly less after 12 weeks, but not different from the fractures stripped of periosteum after 4 and 8 weeks. Bending moment increased throughout the experimental period in both groups. In the fractures with attached periosteum, bending moment was significantly larger at 12 weeks and bending rigidity after 4 weeks. No differences were found in total bone blood flow. Blood flow in the segmental fractured area was substantially increased in both groups after 4 weeks. At 8 weeks the segmental flow was significantly larger in the fractures with stripped periosteum, while a normalization was observed in those with attached periosteum. This study indicates that segmental fractures initiate a substantial increase in blood flow, and that additional stripping of the periosteum delays normalization of the flow. Periosteal stripping of the segment impairs fracture healing, measured as gain of bending moment. PMID- 8723408 TI - Irreducible fracture of the calcaneus due to flexor hallucis longus tendon interposition. AB - An unusual fracture-dislocation of the calcaneus is presented in which tendinous interposition in the subtalar joint prevented the possibility of closed reduction. This situation has been reported in the literature in only five previous patients. The position of the flexor hallucis tendon in the joint could be visualized on the preoperative computed tomographic scan. The patient was treated operatively with reduction through a lateral approach without a supplemental medial incision. PMID- 8723409 TI - Hazards to the orthopaedic trauma surgeon: occupational exposure to HIV and viral hepatitis (a review article). AB - The risk of transmission of blood-borne pathogens is directly related to the prevalence of the infectious disease in the physician's patient population, the infectivity after a single exposure, and the frequency and nature of exposure. After reading this material the reader should understand the guidelines for risk prevention of human immunodeficiency virus (HIV), hepatitis B virus, and hepatitis C virus transmission as published by the Centers for Disease Control and Prevention (CDC) and the Occupational Safety and Health Administration (OSHA) and be better able to evaluate and choose personal protective devices for use while caring for patients. To date there have been no documented cases of HIV seroconversion from solid-bore needle puncture or aerosolized transmission from an HIV-positive patient to a physician in the operating room. However, the authors and the Orthopaedic Trauma Association urge all physicians who have sustained an occupational seroconversion to communicate with the CDC so that prevention guidelines and health-care worker protection can be improved. Regardless, all physicians performing invasive procedures must reevaluate their techniques to determine what procedures can be modified or new instruments developed to reduce the risk of exposure. PMID- 8723410 TI - Are cytokine gene polymorphisms important in the pathogenesis of inflammatory bowel disease? PMID- 8723411 TI - Ulcerative colitis: how can we test theories of immune dysregulation. PMID- 8723412 TI - Evidence of genetic heterogeneity in IBD: 1. The interleukin-1 receptor antagonist in the predisposition to suffer from ulcerative colitis. AB - OBJECTIVE: To investigate a polymorphism within intron 2 of the interleukin-1 receptor antagonist (IL-1ra) gene in a Dutch white population of patients with ulcerative colitis and Crohn's disease. DESIGN: Genotype and allele frequencies of the polymorphic region in the IL-1ra gene were determined in 111 unrelated patients with ulcerative colitis, 92 patients with Crohn's disease, and 86 healthy controls. METHODS: The polymorphic region on the IL-1ra gene was amplified by the polymerase chain reaction (PCR). The resultant products were analyzed by electrophoresis on 2% agarose gels and stained with ethidium bromide. Amplification of the second exon of HLA-DRB1, HLA-DQA1, and HLA-DQB1 genes was performed by PCR, and Dot-blot analysis with biotin-labelled sequence-specific oligonucleotide probes was used for HLA typing. The standard perinuclear antineutrophil cytoplasmic antibodies (pANCA) indirect immunofluorescence assay was performed according to the protocol described by the First International Workshop on ANCA. RESULTS: The frequency of allele 2 of the IL-1ra gene in ulcerative colitis (27.0%) and Crohn's disease patients (25.5%) did not differ significantly from healthy controls (23.8%). However, the estimate of the relative risk for carriers of allele 2 for ulcerative colitis [odds ratio (OR): 1.35, 95%-confidence interval (CI) from 0.73 to 2.49] was in agreement with a previous British study. The exact test for homogeneity of odds ratios provided no evidence for heterogeneity between both populations (P = 0.35) and therefore confirmed the genetic associated between ulcerative colitis and the IL-1ra allele 2 in a different population. Our data confirm that, in ulcerative colitis, the presence of this allele is a genetic marker for severity of the disease. A significant association was demonstrated between the carriership of allele 2 of the IL-1ra gene and the trend over the three localizations in ulcerative colitis (P = 0.023). The same approach for Crohn's disease when compared with healthy controls did not provide evidence for a similar association. The meta-analysis, based on the British data and our data, yielded: OR = 1.06, 95%-CI from 0.71 to 1.59, and P = 0.767. No association between IL-1ra gene polymorphism, and pANCA and the HLA-DR2 allele was found in ulcerative colitis. PMID- 8723413 TI - Absence of antineutrophil cytoplasmic antibodies in relatives of UK patients with primary sclerosing cholangitis and ulcerative colitis. AB - OBJECTIVE: Perinuclear antineutrophil cytoplasmic antibodies (ANCA) have been reported in patients and relatives of patients with ulcerative colitis and primary sclerosing cholangitis, suggesting that ANCA may be a genetic marker of disease susceptibility. The reported frequency of ANCA in relatives has varied greatly, between 0 and 30%. We therefore studied the prevalence of ANCA in unaffected first-degree relatives of British patients with primary sclerosing cholangitis and ulcerative colitis. DESIGN: Thirty-six patients with ulcerative colitis, 33 with primary sclerosing cholangitis and 187 relatives were studied. Ninety-seven relatives were from the primary sclerosing cholangitis proband and 90 were from the ulcerative colitis proband. As an environmental control, 32 spouses were included: 14 from the primary sclerosing cholangitis group and 18 from the ulcerative colitis group. Eighteen healthy volunteers were additional controls. METHODS: ANCA was detected using immunoalkaline phosphatase method. RESULTS: Only 3 of 97 (3%) of the primary sclerosing cholangitis proband relatives had ANCA. One of these had ulcerative colitis, one had rheumatoid arthritis and the third systemic lupus erythematosus. Both rheumatoid arthritis and system lupus erythematosus are known to exhibit ANCA. All other sera were negative. CONCLUSION: ANCA was found only in patients with primary cholangitis and ulcerative colitis and not in their healthy first-degree relatives. ANCA is therefore not a genetic marker for increased disease susceptibility to primary sclerosing cholangitis or ulcerative colitis in the British population. PMID- 8723414 TI - Serum 7 alpha-hydroxy-4-cholesten-3-one and selenohomocholyltaurine (SeHCAT) whole body retention in the assessment of bile acid induced diarrhoea. AB - OBJECTIVE: To assess the reliability of serum 7 alpha-hydroxy-4-cholesten-3-one (7 alpha-3ox-C) in the differential diagnosis of bile acid induced diarrhoea by comparison with 75selenohomocholyltaurine whole body retention (SeHCAT WBR). DESIGN: One hundred and sixty-four patients with chronic diarrhoea were investigated prospectively in two centres (Edinburgh and Sweden) by two different tests which measure bile acid loss or synthesis: the SeHCAT test which measures the 7-day SeHCAT WBR and serum 7 alpha-3ox-C which reflects the rate of bile acid synthesis. RESULTS: Forty-six patients had SeHCAT WBR of less than 10% (19 with ileal disease or resection, nine with idiopathic bile acid induced diarrhoea and 18 with miscellaneous causes for bile acid induced diarrhoea). All patients with ileal or idiopathic disease showed a favorable response to treatment as did 13 of the miscellaneous group. Serum 7 alpha-3ox-C was raised in all subjects with ileal disease/resection, seven patients with idiopathic disease and all subjects in the miscellaneous group who responded to treatment. Sixteen out of 118 patients with SeHCAT WBR greater than or equal to 10% had raised serum 7 alpha 3ox-C. CONCLUSION: The positive predictive value of serum 7 alpha-3ox-C was 74%. The high negative predictive value (98%) of serum 7 alpha-3ox-C indicates the possible use of this test for excluding bile acid malabsorption in this population. All but two subjects who responded to treatment had raised serum 7 alpha-3ox-C concentrations. The possibility that the sensitivity of the test can be improved by repeat testing needs to be further investigated. There was a significant correlation between fractional catabolic rate (FCR) SeHCAT and serum 7 alpha-3ox-C (r = 0.63, P < 0.0001). Further data are required to validate the reference range in women over 70 years of age. PMID- 8723415 TI - Liver steatosis and chronic hepatitis C: a spurious association? AB - OBJECTIVE: Based on the observation of steatosis in the majority of liver biopsy specimens from hepatitis C virus (HCV) infected patients, it has been suggested that HCV may be pathogenetically implicated. We aimed to determine the influence of possible underlying metabolic disorders on this association. DESIGN: In a series of 148 consecutive patients with chronic hepatitis, with and without HCV infection, we evaluated by logistic regression analysis the association between steatosis and HCV, controlling for diabetes, obesity, hyperlipidaemia and alcohol. These are all known to be factors associated with a fatty liver, and also with the histological degree of liver disease. RESULTS: Antibodies to HCV were detected in 121 of 148 (81.8%) patients. Steatosis, distributed in different histological patterns, was found in 73 of 121 (60%) HCV-positive and in 14 of 27 (52%) HCV-negative patients (P = NS). Using simple logistic regression, the association Of HCV to steatosis was weak and not statistically significant (OR, 1.14; 95% CI, 0.61-3.27). The same was true for hyperlipidaemia (OR, 4.45; 95% CI, 0.52-37.9). A strong and statistically significant association was found, however, between obesity and steatosis (OR, 4.18) and between steatosis and the highest degree of histological severity (Liver cirrhosis vs chronic persistent hepatitis: OR, 12.8). Using multivariate analysis, the association between steatosis and HCV was shown to be not significant. Hyperlipidaemia, among all the independent variables tested, was shown to be co-linear with obesity. CONCLUSION: Our findings seem to suggest that HCV is irrelevant as a risk factor for a fatty liver. The results indicated that there is a 'confounding' role of obesity and hyperlipidaemia, and that the severity of liver disease is associated with steatosis and HCV. PMID- 8723416 TI - Bile levels of carcino-embryonic antigen in patients with hepatopancreatobiliary disease. AB - OBJECTIVE: To evaluate the value of biliary carcino-embryonic antigen (CEA) in the differential diagnosis of malignant and benign hepatopancreatobiliary disease. PATIENTS: One hundred patients were prospectively studied. Benign diseases were present in 39% of the patients while 61% had malignant diseases. METHODS: Samples of serum were taken from all patients just before endoscopic retrograde cholangiopancreatography (ERCP) and samples of biliary CEA were obtained during ERCP. RESULTS: The sensitivity of serum CEA and carbohydrate antigen 19-9 (CA 19-9) in detecting malignancy were 50% and 92%, respectively, while the respective specificities were 95% and 72%. The mean biliary CEA level of the benign group was significantly different from that of the malignant group (35.7 +/- 8.7 ng/ml vs 268 +/- 85.5 ng/ml), but there was considerable overlap between the two groups. With a cut-off level of 20 ng/ml, the sensitivity and specificity were 84% and 64% respectively. The mean bilirubinaemia value was significantly higher in malignant disease than in benign disease (57.4 +/- 13.9 mumol/l vs 235 +/- 19.8 mumol/l). Multidimensional analysis indicated that only bilirubinaemia (P < 109-3)) was independently predictive of malignant disease. CONCLUSION: Biliary CEA assessment seems useless in distinguished between benign and malignant causes of cholestasis. PMID- 8723417 TI - Neutrophil activation in chronic liver disease. AB - OBJECTIVE: To assess the relationship between neutrophil activation and indices of disease severity in patients with chronic liver disease. METHODS: Plasma neutrophil elastase was measured by radioimmunoassay as a marker of neutrophil activation, and disease severity assessed by standard clinical, biochemical, haematological and histological techniques. PATIENTS: Eighty-eight patients with chronic liver disease were studied, Thirty-nine had alcohol-induced liver disease (ALD), 18 autoimmune chronic hepatitis, 13 cryptogenic cirrhosis, seven primary biliary cirrhosis, six primary sclerosing cholangitis, three haemochromatosis and two secondary biliary cirrhosis. Seventy-three patients were cirrhotic and 15 were non-cirrhotic, confirmed by biopsy. RESULTS: Levels of neutrophil elastase were raised in Childs C cirrhotic patients with ALD compared with Childs A or B patients with ALD (P < 0.01), Childs A or B patients with non-ALD (P < 0.01), and Childs C patients with non-ALD (P = 0.02). In patients with ALD, neutrophil elastase correlated with prothrombin time (r = 0.679, P = 0.001), bilirubin (r = 0.587, P < 0.001), Child-Pugh score (r = 0.546, P < 0.001) and inversely with serum albumin (r = -0.511, P < 0.001). In patients with non-ALD, there were no correlations with the measurements of with transaminase levels. CONCLUSION: Neutrophil activation, as measured by plasma neutrophil elastase, is a marker of disease severity in patients with alcohol-induced chronic liver damage, but not in those with other causes of liver disease. PMID- 8723418 TI - Carcinoma of the ampulla of Vater: a tumour with a poor prognosis? AB - OBJECTIVE: To assess the overall prognosis of patients with ampullary carcinomas and evaluate the presentation, diagnosis, pathology and management of these potentially highly curable tumours, attempting to relate these factors to overall survival. PATIENTS AND METHODS: Forty patients with ampullary carcinoma were reviewed. Age, Sex, nature and duration of history, laboratory information at admission, results of diagnostic radiology, endoscopic retrograde cholangiopancreatograms and pathological findings were considered. Both curative and palliative management strategies were reviewed. RESULTS: The overall median survival was 19 months. The median duration of history was 5.9 +/- 5.4. weeks, with no significant difference in survival between patients with short and those with long histories (P = 0.46). Twenty nine (73%) patients were potentially resectable, but only 15 (37%) underwent potentially curative surgery. The difference in survival between the Whipple's (13) and the endoscopically stented (20 ) groups was not significant (p = 0.08). The Whipple's group were significantly younger than the stented group (P = 0.001) and had a significant operative morbidity, re-operation rate (38%) and post-operative mortality (15%). Only five of 13 patients were alive following Whipple's treatment after a mean follow-up of 18.9 months. Sphincterotomy before Whipple's treatment improved survival significantly (P = 0.04); absence of jaundice, exophytic macroscopic appearance, well-differentiated tumours and early stage were also associated with good survival. Endoscopic retrograde cholangiopancreatography has a high diagnostic yield and a low associated morbidity and mortality, with endoscopic papillectomy aiding cannulation while effective palliation was provided through stenting, endoscopic papillectomy and laser debulking of obstructing tumours. Little benefit was obtained from chemoradiotherapy. CONCLUSION: Despite the potential for curative resection in patients with ampullary carcinoma, the majority of such patients are unsuitable for curative surgery on grounds of age, general health status or advanced disease; since only 37% of patients undergo potentially curative surgery the condition has a poor prognosis. PMID- 8723419 TI - ERCP: present practice in a single region. Suggested standards for monitoring performance. AB - A survey of consecutive endoscopic retrograde cholangiopancreatography (ERCP) procedures has been undertaken for all the Units performing this procedure in the (old) Northern region of the UK. The process, imaging success, procedural success and complications have been analysed for 255 ERCP procedures. Overall, the results seem to be reassuring, with an imaging success rate of 88% (n = 297 ducts required), additional procedural success rate of 88% (n = 178) and with a serious complication rate of 6%. However, there are wide differences both in terms of practice (contrast agent used, antibody prophylaxis), imaging success (76-95%) and outcomes (procedure success 75-100%, serious complications 0-16%). This wide difference in practice seems difficult to justify. Individual Units should monitor their performance. Standards of practice are suggested for discussion and debate. PMID- 8723420 TI - Sigmoidoscopy and rectal biopsy: a survey of current UK practice. AB - OBJECTIVE: To establish the pattern of practice of sigmoidoscopy and rectal biopsy in the UK, and to estimate the rate of occurrence of important complications. DESIGN: Postal questionnaire enquiring about practice in the previous 2 years. SUBJECTS: Consultant members of the British Society of Gastroenterology. MAIN OUTCOME MEASURES: Bleeding or perforation following sigmoidoscopy and rectal biopsy. RESULTS: Completed replies were received from 277 consultants, giving a response rate of 84%. Of the sigmoidoscopies performed, 71% were rigid and the remainder fibreoptic flexible sigmoidoscopy (FOS). The complication rate after sigmoidoscopy and rectal biopsy was 0.01%. Perforation was significantly more common in FOS than in rigid sigmoidoscopy (eight in 134,482 sigmoidoscopies versus five in 328,815, chi2 = 5.18, P < 0.05). The distance from anal margin and wall of biopsy did not influence perforation rates. Significant bleeding was similar with both techniques (eight in 134,482 versus 35 in 328,815, chi2 = 2.27, NS). CONCLUSION: Rigid sigmoidoscopy is still more widely used than FOS in the UK, and is probably safer. Complications resulting from rectal biopsy are uncommon but perforation occurs more frequently with FOS than with rigid sigmoidoscopy. The site of biopsy appears to be unimportant. PMID- 8723421 TI - Helicobacter pylori, pepsinogens and gastrin: relationship with age and development of atrophic gastritis. AB - BACKGROUND: Helicobacter pylori causes chronic gastritis in all infected individuals and thus may be a risk factor for the ultimate development of trophic gastritis and gastric cancer. The serum levels of pepsinogen A, pepsinogen C and gastrin can be used as markers for both non-atrophic and atrophic gastritis. METHODS: We determined the serum levels of gastrin, pepsinogen A and pepsinogen C and the pepsinogen A/C ratio in 150 H. pylori-negative and 186 H. pylori-positive individuals. RESULTS: The H. pylori infected patients had significantly higher serum levels of pepsinogen A, pepsinogen C and gastrin and a significantly lower pepsinogen A/C ratio. In the non-infected patients, none of the respective serum values changed with increasing age. In contrast, in the infected patients, the pepsinogen A level and pepsinogen A/C ratio decreased significantly with increasing age. CONCLUSION: H. pylori infection increases serum levels of pepsinogen A, pepsinogen C and gastrin and decreases the pepsinogen A/C ratio. In infected subjects, levels of pepsinogen A and the pepsinogen A/C ratio decrease with ageing. These findings support the concept of H. pylori as a risk factor for the development of atrophic gastritis. PMID- 8723422 TI - Chronic Diarrhoea: current concepts on mechanisms and management. PMID- 8723423 TI - Plasma cell variant of Castleman's disease localized to the liver: a case report. AB - Localized plasma cell variant of Castleman's disease is an uncommon lymphoproliferative disorder. It is characterized by numerous, evenly distributed germinal centres with moderate or extensive sheets of plasma cells in between. It may be associated with a wide variety of systemic manifestations that disappear after removal of lesion. Rare cases of Castleman's disease have been reported in extranodal locations, i.e. muscle, larynx, vulva, lung, pericardium and cranium. We report the first the case of a liver tumour with clinico-pathological features of the plasma cell variant of Castleman's disease. This tumour differs from the hepatic inflammatory pseudotumour, which is frequently associated with infectious conditions. PMID- 8723424 TI - Treatment of a symptomatic hepatic cyst by tetracycline hydrochloride instillation sclerosis. AB - We report the case of an 84-year-old woman with abdominal pain, distension and inferior veno-caval compression due to a large non-parasitic hepatic cyst. She was treated by instillation of tetracycline hydrochloride into the hepatic cyst. Six months after treatment, the patient is asymptomatic, with no reaccumulation of the hepatic cyst. We conclude that symptomatic hepatic cysts may be successfully treated by tetracycline hydrochloride instillation sclerosis. PMID- 8723425 TI - Idiopathic mesenteric varices causing lower gastrointestinal bleeding. AB - We report a case of lower gastrointestinal bleeding caused by idiopathic mesenteric varices. A 25-year-old man presented with a history of two episodes of lower gastrointestinal bleeding without transfusion. Colonoscopy revealed varices of the entire colon and terminal ileum. Intravariceal blood flow was demonstrated by dopplersonography. Vascular abnormalities were excluded by selective angiography of the upper and lower mesenteric artery. No therapy was given without a new episode of bleeding and there has been no further bleeding to date (a period of 14 months). Mesenteric varices are a rare cause of lower gastrointestinal bleeding, almost always associated with portal hypertension. The varices are idiopathic in only a few cases. The therapy depends on the intensity of bleeding, but resection is the most frequent treatment. PMID- 8723426 TI - Long-term outcome of idiopathic hypereosinophilic syndrome--transition to eosinophilic gastroenteritis and clonal expansion of T-cells. AB - We describe a patient with idiopathic hypereosinophilic syndrome, without initial gastrointestinal symptoms, and their transition to eosinophilic gastroenteritis. This patient, a 65-year-old man, presented with fever, constitutional symptoms, peripheral and bone marrow eosinophilia 20 years ago. During the course of the disease, diarrhoea and malabsorption became prominent, whereas bone marrow eosinophilia regressed completely and blood eosinophilia regressed partially. Biopsies showed a severe eosinophilic gastroenteritis of the mucosal type involving the stomach, small bowel and colon. During the final years of the patient's disease, mucosal eosinophilia became less intense and a mucosal infiltration with T-cells dominated. At autopsy, immunopathological studies of small intestines and colon specimens showed a clonal expansion of morphologically normal T-cells in the intestinal mucosa, which expressed the abnormal phenotype CD2+CD3+CD4-CD5-CD8-. Flow cytometry examination of peripheral blood revealed a corresponding abnormal population of CD3+CD4-CD8- T-cells, indicating a systemic spread of the process. The patient eventually died of non-obstructive small bowel infarction with peritonitis 20 years after the onset of the first symptoms. We postulate that the destructive eosinophilic/lymphocytic inflammation is caused by a clonal proliferation of T-lymphocytes with probable secretion of Type 2 T(helper) cell cytokines and consecutive stimulation of eosinophils. PMID- 8723427 TI - Xerostomia: diagnosis and management. AB - Xerostomia, or dry mouth, is a common complaint that may be caused by several conditions, which include side effects of a wide variety of drugs, such as antidepressants, therapeutic radiation to the head and neck, dehydration, diabetes, and diseases involving salivary glands, such as Sjogren's syndrome. The complaint of dry mouth may or may not be associated with decreased salivary gland function. Individuals with xerostomia complain of problems with eating, speaking, swallowing, and wearing dentures. Some people also complain of salivary gland enlargement or changes in taste. Lack of saliva may predispose one to oral infections, such as candidiasis, and increase the risk of dental caries. Management of the individual patient with xerostomia includes assessment of salivary gland function, replacement therapy, and prevention of caries and oral candidiasis. Early recognition and management of xerostomia may prevent devastating dental disease and help to improve the quality of life. PMID- 8723428 TI - A nursing perspective on radiation-induced xerostomia. AB - Xerostomia during and following a course of head and neck irradiation profoundly impacts the quality of life of many patients. Xerostomia not only affects mucous membranes and teeth but also interferes with patient comfort, nutrition, and activities of daily living. A thorough evaluation of xerostomia is essential and should include providing anticipatory guidance to the patient and family. In addition, education on prophylactic oral care is necessary during the initial phases of treatment. As symptoms occur, various palliative interventions are tailored to the patient's and family's needs, promoting adherence to mouth care regimens and enhancing patient comfort. Long-term follow-up with education and counseling is critical for optimal patient management. PMID- 8723429 TI - A multicenter maintenance study of oral pilocarpine tablets for radiation-induced xerostomia. AB - Two hundred sixty-five patients with head and neck cancer who had previously participated in either a fixed-dose, dose-titration, or dose-ranging trial of oral pilocarpine hydrochloride tablets were enrolled in a 36-month multicenter maintenance study to evaluate the long-term safety and efficacy of oral pilocarpine for the treatment of radiation-induced xerostomia. In this open-label study, the initial drug dose was 5.0 mg tid, with possible adjustments from 2.5 to 10.0 mg tid or bid. Efficacy was evaluated by subjective measures of oral function. Safety evaluations were based on self-report of symptoms (or of adverse effects), various examinations, and laboratory tests. There was significant improvement in all criteria of oral function. Sweating was the most frequent adverse experience (55%). Less frequent side effects, mild to moderate in nature, included increased urinary frequency, lacrimation, and rhinitis. Side effects usually diminished within hours after the cessation of therapy. We conclude that oral pilocarpine at these doses effectively and safely reduces the symptoms of radiation-induced xerostomia. PMID- 8723430 TI - Effects of long-term psychological stress on sexual behavior and brain catecholamine levels. AB - The effects of long-term psychological stress on sexual behavior and brain catecholamines were investigated in rats. Stress was applied using the communication box developed by Ogawa and Kuwabara (1996), and a psychological stress group (n = 12), a physical stress group (n = 5), and a control group (n = 5) were established. Stress was applied for 1 hour every day for 10 consecutive weeks. Sexual behavior was observed before the start of exposure to stress and 2, 4, 6, 8, and 10 weeks thereafter. The results showed that long-term psychological stress impaired the sexual behavior of male rats. Long-term psychological stress decreased the concentrations of catecholamine and its metabolites in the brain, especially in the medial preoptic area (MPOA). Thus, we hypothesized that low catecholamine neurotransmission in the brain results in impairment of male rat sexual behavior. We then tried to restore the impaired sexual behavior by administration of a cerebral-activating drug, indeloxazine hydrochloride. The administration of indeloxazine hydrochloride for a 3-week period restored the sexual behavior that had been impaired by long-term psychological stress. These present results suggest that impairment of neurotransmission in the central nervous system could be a cause of sexual dysfunction, and activation of neurotransmission may result in restoration of impaired male sexual behavior. PMID- 8723431 TI - Morphometric study of the gubernaculum in male estrogen receptor mutant mice. AB - To determine role of estrogen receptors in testicular descent, a morphometric study of the testis and structures derived from the gubernaculum was made in sexually mature male mice having an estrogen receptor disrupted gene mutation (ERKO). Macroscopic dissections and sagittal serial sections were made of the pelvis of four wild-type mice, four mice heterozygous for the ERKO mutation, and four homozygous ERKO males. By external morphological examination the testes appeared to be descended in all three genotypes. All mice had development of a cremaster sac, which is derived from the gubernaculum, but this was twice as large in wild-type mice than in both the heterozygote or homozygote ERKO groups. The cause for the smaller cremaster sac appeared to be excessive development of the cremaster muscle in ERKO mice. The thickened muscle was associated with postmortem retraction of the testes into the inguinal canal or abdomen. Spermatogenesis and testicular volume were deficient in homozygous ERKO mice at this age. This study demonstrates that estrogen has a previously unknown role in masculine sexual development of the gubernaculum and the structures derived from it, such as the cremaster muscle. PMID- 8723432 TI - Ultrastructural and immunohistochemical characterization of the tunica albuginea in Peyronie's disease and veno-occlusive dysfunction. AB - The tunica albuginea and corpus cavernosum from patients with Peyronie's disease (PD), patients with veno-occlusive dysfunction (VOD), and those from normal control subjects were studied by transmission electron microscopy and immunohistochemical staining for type I, III, and V collagens, platelet-derived growth factor (PDGF) AA and BB homodimers, and PDGF alpha and beta receptors. Ultrastructural modifications resembling a fibrotic reaction were detected in the two pathological tunica albuginea, but not in those from control subjects. Ultrastructural data demonstrated a general increase in fibrous and amorphous extracellular matrix material in the pathological tunica albuginea. The amorphous material probably represents glycoproteins and proteoglycans. The fibrous material, representing collagen, appears disorganized in the tissue and does not display the typical and homogeneous diameter, size, and spatial arrangement. Large areas of extracellular and intracytoplasmic, partially degraded, fibers are visible. An increased type I/III collagen ratio was detected by immunohistochemistry in the two pathological tunica albuginea. Moreover, a strong expression of type V collagen, correlated to fibroblasts, was revealed. Fibroblasts from control tissues, on the other hand, were totally negative. Finally, PDGF AA and BB were positive in fibroblasts from pathological tunica albuginea but were negative in control tissues. PDGF beta receptor was positive in pathological and normal tissue fibroblasts. Tunica albuginea from PD and VOD show similar ultrastructural and immunohistochemical alterations, whereas the corpus cavemosum shows no visible modifications. PMID- 8723433 TI - Regulation of the epididymal synthesis of P26h, a hamster sperm protein. AB - We have previously identified a 26-kDa epididymal hamster sperm glycoprotein (P26h) that is involved in gamete interaction. This protein is added to the acrosomal cap during epididymal transit of spermatozoa. Because the epididymis secretes proteins under androgen control, the aim of this study was to document the testicular control of the epididymal ontogenesis of P26h. The cytosolic fraction of the epididymides of male hamsters of different ages, prepared by ultracentrifugation, was used as well as those from mature males at different times following castration. These extracts were Western blotted and probed with an anti-P26h antiserum. P26h was initially immunodetectable in extracts prepared from hamsters 4 weeks of age, and the signal increased up to the 7th week of age. The P26h signal decreased rapidly after castration until the antigen was undetectable at 3 days following castration. Administration of testosterone to 3 week-old male hamsters resulted in the epididymal expression of P26h earlier than that observed in untreated or control animals. These results suggest that P26h protein expression in hamster epididymis is under testicular androgen control. PMID- 8723434 TI - Mouse epididymal sperm contain active P450 aromatase which decreases as sperm traverse the epididymis. AB - Recently we reported that mouse germ cells in the testis contain active P450 aromatase (P450arom), the enzyme that converts androgens to estrogens. This finding suggested that germ cells have the ability to produce estrogen. Further studies have shown that germ cells in the testis of several species contain P450arom. The goal of this study was to determine if epididymal sperm contain P450arom and if P450arom activity in sperm changes during traversion of the epididymis in the adult mouse. P450arom was localized in sperm present in the efferent ductules and epididymis by immunocytochemistry using an antiserum generated against purified human placental cytochrome P450arom. P450arom immunostaining in sperm was most prominent in sperm located in the proximal caput epididymis, decreased as sperm traversed the corpus epididymis, and was only slightly apparent in sperm in the cauda epididymis. The immunolocalization of P450arom in epididymal sperm was supported by the measurement of P450arom activity in sperm by the 3H2O assay. We found that P450arom activity in sperm significantly decreases as sperm traverse the epididymis. Based upon these observations, we conclude that sperm can synthesize estrogen and that the synthesis of estrogen by sperm present in the efferent ductules and caput epididymis could be important in the process of sperm maturation. PMID- 8723435 TI - Changes in luminal fluid protein composition in the rat cauda epididymidis following partial sympathetic denervation. AB - Sympathetic denervation of the rat cauda epididymidis by surgical removal of the inferior mesenteric ganglion (IMG) results in an excessive accumulation of sperm in the cauda epididymidis as well as significant changes in cauda sperm motility and cauda epididymal gross histology. The objective of the present study was to determine if the cauda-specific changes in sperm storage, sperm motility, and epididymal histology following the loss of sympathetic innervation were accompanied by changes in the protein composition of epididymal fluid. One and 4 weeks after surgical IMG removal or sham operations, luminal fluid obtained from the caput and cauda epididymidis and cauda epididymal sperm-associated proteins were subjected to two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) and silver-stained proteins were quantitated. One week after IMG removal, two cauda epididymal fluid (CEF) proteins (2 and 13) had increased 43% and 49%, respectively, whereas four CEF proteins (5, 8, 9, and 19) had decreased between 30% and 73% compared to controls. Four weeks after IMG removal, changes in CEF proteins observed 1 week following surgery were no longer present, but the staining intensities of three additional CEF proteins (11, 12, and 18) were reduced an average of 70% compared to control CEF proteins. By obstructing the cauda epididymidis, we confirmed that the changes in CEF protein composition observed following IMG removal were not the result of sperm accumulation but were due directly to the loss of innervation; the staining intensity of CEF protein 2 increased as a result of excessive sperm accumulation in the cauda epididymidis both in the presence and absence of innervation from the IMG. No significant changes in caput epididymal fluid proteins or cauda epididymal sperm-associated proteins were detected following IMG removal. These data show that the protein composition of rat CEF is significantly affected by the loss of sympathetic innervation and suggest that neuronal input may play an important role in the maintenance of epididymal function. PMID- 8723436 TI - Detection of germ cell-derived proteins in testicular interstitial fluid: potential for monitoring spermatogenesis in vivo. AB - The aim of the present study was to assess whether proteins secreted by the seminiferous tubules (ST) can be detected in testicular interstitial fluid (IF) and testicular (TV), spermatic (SV), and peripheral venous (PV) plasma from adult rats. An antiserum was raised against seminiferous tubule-conditioned medium (STCM) prepared from adult rats and used in conjunction with Western blot analysis to screen IF and blood samples resolved by one-dimensional (1-D) and two dimensional (2-D) sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS PAGE). Samples of IF and PV were analyzed from control adult rats and rats exposed to scrotal heating (43 degrees C for 30 minutes) 24 hours earlier to ascertain whether damage to spermatogenesis would affect 'leakage' of proteins from the seminiferous tubules. In all control rats, the STCM antiserum specifically detected three proteins in testicular IF with molecular weights of 24, 16, and 14 kDa, respectively. Heat treatment increased the abundance of these proteins and induced the appearance of several other less-abundant proteins, all with molecular masses below 25 kDa. Two of the proteins present in IF were identified, the 24-kDa protein as phosphatidylethanolamine-binding protein (PEBP), and the 14-kDa protein as an androgen-regulated protein (ARP-2). Both of these proteins have been shown in previous studies to be secreted by round spermatids. Our results suggest that germ cell secretory products can gain access to the interstitium under both normal physiological conditions and more easily after induction of damage to spermatogenesis. The antiserum was unable to detect any ST-derived proteins in blood, although it is likely that this result may be due to insensitivity of the presently used techniques. The development of specific immunoassays for germ cell-secreted proteins (e.g., PEBP and ARP-2) should enable more definitive assessment of whether proteins secreted by the seminiferous epithellum can be measured in blood and thus provide a potential means of monitoring spermatogenesis. PMID- 8723437 TI - Testosterone autoregulation of its biosynthesis in the rat testis: inhibition of 17 alpha-hydroxylase activity. AB - Using the in vitro perfused rat testis, the effects of testosterone (T) on its own biosynthesis, and in particular on the inhibition of specific steroidogenic step(s) in the biosynthetic pathway from cholesterol to T, were examined. Rat testes perfused in vitro for 1 hour with medium containing 1.5 microM T secreted significantly less T than control testes in response to physiologic or maximal luteinizing hormone (LH) stimulation. To locate the site(s) of this rapid inhibition, the effects of arterial T infusion on steroidogenesis by testes also infused with pregnenolone (PREG), progesterone (PROG), 17 alpha hydroxyprogesterone (17-PROG), or androstenedione (ADIONE) were measured by summing all the possible reaction products from each substrate. This approach allowed us to examine the effect of T in situ on the reactions: LH-stimulated PREG secretion; PREG to PROG; PROG to 17-PROG; 17-PROG to ADIONE; and ADIONE to T. Only PROG to 17-PROG (17 alpha-hydroxylase activity) was inhibited by arterial T infusion. A kinetic examination of the PROG to 17-PROG reaction demonstrated that the specific inhibition by T was competitive. The apparent km for PROG in this system was 16.0 microM, whereas the apparent ki of T was 1.6 microM, indicating a relatively high degree of sensitivity of the reaction to T. Taken together, these data confirm that T is able to regulate its own synthesis and indicate that this autoregulation is the result of rapid, specific inhibition by T of 17 alpha-hydroxylase activity. PMID- 8723438 TI - Hexokinase present in human sperm is not tyrosine phosphorylated but its antibodies affect fertilizing capacity. AB - The present study was conducted to investigate the presence of hexokinase in human sperm cell, its subcellular localization, its modulation and role in capacitation/acrosome reaction, and tyrosine kinase activity, if any. These studies were conducted using antibodies (Ab) raised against rat brain hexokinase (type I isozyme) that have significant cross-reaction with various human tissue hexokinases and neutralize the catalytic activity of the enzyme. Hexokinase Ab reacted with acrosomal, mid-piece and tail regions of methanol-fixed (approximately 70-80%) and live (approximately 35-52%) sperm in the indirect immunofluorescence technique (IFT) and the immunobead binding technique (IBT), respectively. Hexokinase Ab specifically recognized a band of 116 kDa on the Western blot of detergent-solubilized human sperm preparation that was different from the 95-kDa phosphotyrosine protein. Hexokinase Ab caused a significant (P < 0.01 to < 0.001) and concentration-dependent inhibition of human sperm penetration of zona-free hamster oocytes in the sperm penetration assay (SPA). These data indicate that the hexokinase of 116-kDa molecular weight is present in acrosomal, mid-piece, and tail regions of human sperm. The sperm hexokinase is a glycoprotein that is different from the 95-kDa phosphotyrosine protein and is not phosphorylated at tyrosine residues; however, its antibodies cause agglutination and a concentration-dependent inhibition of fertilizing capacity of human sperm. PMID- 8723439 TI - Effect of sperm lipid peroxidation on fertilization. AB - This study was aimed at determining, with homologous mouse gametes, whether a level of membrane lipid peroxidation insufficient to affect sperm motility parameters can alter sperm fertilizing potential. The addition of ferrous ions and ascorbic acid (Fe2+/ Asc) to mouse sperm suspensions increases the formation of thiobarbituric acid-reactive substances (TBARS), an indicator of lipid peroxide breakdown products, without significantly affecting the level of sulfhydryl groups. In the presence of Fe2+/Asc concentrations above > 0.4/2.0 mM, spermatozoa become immotile. However, at concentrations < or = 0.4/2.0 mM of Fe2+/Asc, i.e., conditions in which the TBARS formation is increased by < or = 4.6-fold over that of controls, motility remains unaffected for up to 3 hours. In the presence of 0.4/ 2.0 mM Fe2+/Asc, treated spermatozoa increase their fertilizing potential by 50%, as measured by the formation of two-cell embryos. This increase is not caused by improvements in sperm motility parameters (percentage, linearity, velocity, hyperactivation) or sperm capacitation. On the other hand, there is a significant increase in the capacity of mouse spermatozoa to bind to homologous zona pellucida. In conclusion, mild peroxidative conditions, that increase lipid peroxide formation 4.6-fold without significantly modifying free sulfhydryl groups and sperm motility parameters, improve the fertilizing potential of spermatozoa by increasing their binding capacity to zona pellucida. PMID- 8723440 TI - Cytokine levels in the seminal plasma of infertile males. AB - Cytokines released by various cell subsets in the male urogenital tract are capable of markedly influencing sperm function and fertility. We determined the cytokine content in the seminal plasma of patients with unexplained infertility and correlated the results with urogenital infections and sperm parameters. Routine sperm parameters, bacterial culture of seminal plasma and blood follicle stimulating hormone (FSH), luteinizing hormone (LH), and testosterone were obtained from 14 infertile males and 8 healthy control subjects. Interleukin 1 beta (IL-1 beta), interleukin 6 (IL-6), and tumor necrosis factor alpha (TNF alpha) levels in the seminal plasma were measured by enzyme-linked immunosorbent assay (ELISA). IL-1 beta, IL-6, and TNF alpha levels in the seminal plasma were negatively correlated with the number of progressively motile sperm, but there was no correlation with total sperm counts, viability, pH, morphological alterations, type of abnormality, and hormone parameters. Cytokine levels were significantly elevated in seminal plasma exhibiting bacterial or mycoplasmal infections of the urogenital tract. Urogenital infections lead to an release of inflammatory cytokines, most probably by immunocompetent cells of the lymphocyte/macrophage origin. Cytokines such as IL-1, IL-6, and/or TNF alpha might influence sperm motility via direct or indirect effects, resulting in reduced mucosa penetration properties. Therefore, our data suggest that cytokines may be involved in reduced male fertility. PMID- 8723441 TI - Influence of semen collection method on ejaculate characteristics in the common marmoset, Callithrix jacchus. AB - A source of normal spermatozoa (sperm) is required for many andrological and reproductive studies. Ideally the method used for semen collection should be repeatable and reliable but should not influence sperm characteristics or sperm function. Two methods of semen collection from marmosets, vaginal washing after copulation and electroejaculation, were compared in terms of the success rate in obtaining samples and the characteristics of the sperm suspensions. Vaginal washing was shown to be a reliable, repeatable, and apparently non-stressful method of obtaining ejaculates, with 8 out of 10 males ejaculating on at least four of the five attempts. The semen was of good quality as assessed by conventional means, and the sperm were highly fertile when used for artificial insemination (Al; 100% conception rate). In contrast, fewer ejaculates were obtained from the same males by electroejaculation (success rate 30%), sperm survival in vitro was reduced compared to sperm collected by vaginal washing (0.93 +/- 0.15 days for electroejaculated sperm compared to 1.98 +/- 0.3 days for sperm from vaginal washing), and the number of animals giving birth after Al was smaller (0 vs. 6; P < 0.05). The proportions of motile (74.8 vs. 70.7%), live (84.7 vs. 81%), and morphologically normal (91.9 vs. 87.6%) sperm in the ejaculates were not affected by the semen collection method, but velocity parameters, such as curvilinear velocity, straight line velocity, and average path velocity, as assessed by computerized motility analysis, were significantly lower in vaginal washings than in electroejaculates (P < 0.023, 0.008, and 0.008, respectively). Mean angular deviation and beat cross frequency were greater in vaginal washings than in electroejaculates (P < 0.016 and 0.008, respectively). Therefore the effect of semen collection method on sperm function should always be considered when designing reproductive studies. PMID- 8723442 TI - Synthetic combinatorial libraries: novel discovery strategy for identification of antimicrobial agents. PMID- 8723444 TI - Evaluation of a peptidomimetic ribonucleotide reductase inhibitor with a murine model of herpes simplex virus type 1 ocular disease. AB - The ribonucleotide reductase (RR) of herpes simplex virus type 1 (HSV-1) is an important virulence factor, being required for neurovirulence, ocular virulence, and reactivation from latency. The RR activity requires the association of two distinct homodimeric subunits, and the association of the subunits is inhibited in the presence of a peptide homologous to the carboxy terminus of the small subunit. A structural analog of the inhibitory peptide (BILD 1263) has been shown to inhibit the replication of HSV-1 at micromolar concentrations in vitro. We used a mouse model of HSV-1 ocular infection to determine the in vivo efficacy of topical BILD 1263. Treatment of HSV-1 KOS-infected mice resulted in significant reductions in the severity and incidence of stromal keratitis and corneal neovascularization. At higher concentrations (5%) BILD 1263 reduced the severity but not the incidence of blepharitis. Treatment with 5% BILD 1263 also reduced viral shedding from the cornea by 10- to 14-fold (P < 0.001). In uninfected mice treated with 5% BILD 1263, we found no evidence of corneal epithelial damage, conjunctivitis, or blepharitis, and histopathological studies revealed no changes in the corneas of these mice. These results show that the peptidomimetic RR inhibitor BILD 1263 is effective in preventing disease, has an antiviral effect in vivo, and has little or no toxicity. PMID- 8723443 TI - MDL 74,968, a new acyclonucleotide analog: activity against human immunodeficiency virus in vitro and in the hu-PBL-SCID.beige mouse model of infection. AB - The novel acyclonucleotide derivative of guanine, 9-[2-methylidene-3 (phosphonomethoxy)propyl] guanine (MDL 74,968), had antiviral activity comparable to those of 9-(2-phosphonomethoxyethyl) adenine (PMEA) and 2',3'-dideoxyinosine against laboratory strains of both human immunodeficiency virus (HIV) types 1 and 2 cultured in MT-4 cells and several clinical HIV isolates cultured in human peripheral blood mononuclear cells (PBMCs). MDL 74,968 was at least fourfold less toxic than PMEA to MT-4 cells or PBMCs, thereby producing a more favorable in vitro selectivity index for the former compound. Studies of acute toxicity in CD 1 mice showed that MDL 74,968 was not toxic at doses of 1,600 mg/kg of body weight via the intraperitoneal route or at doses of 500 mg/kg via the intravenous route. Furthermore, no adverse effects of MDL 74,968 were apparent when mice were treated at doses of 200 mg/kg twice daily for 5 days. Treatment by continuous subcutaneous infusion of MDL 74,968 or PMEA at the daily dose of 20 mg/kg in the hu-PBL-SCID.beige murine model of HIV infection significantly reduced the severity of infection compared with that in placebo-treated controls. Quantitation of virus recovery by endpoint titration of spleen cells in coculture with mitogen-activated PBMCs demonstrated that MDL 74,968 as well as PMEA significantly reduced the amount of virus (P < 0.02). Moreover, by using DNA extracted from spleens, the mean HIV:HLA PCR product ratio, which takes into account individual variation in immune system reconstitution, were 0.50 and 0.40 for MDL 74,968 and PMEA treatments, respectively, whereas animals receiving the placebo control had significantly higher levels of HIV proviral DNA (mean 0.78; P < 0.02). Taken together, these promising findings suggest that an orally bioavailable prodrug of MDL 74,968 should be developed for the treatment of HIV infection. PMID- 8723446 TI - Population pharmacokinetics of ceftazidime in cystic fibrosis patients analyzed by using a nonparametric algorithm and optimal sampling strategy. AB - Postinfusion data obtained from 17 patients with cystic fibrosis participating in two clinical trials were used to develop population models for ceftazidime pharmacokinetics during continuous infusion. Determinant (D)-optimal sampling strategy (OSS) was used to evaluate the benefits of merging four maximally informative sampling times with population modeling. Full and sparse D-optimal sampling data sets were analyzed with the nonparametric expectation maximization (NPEM) algorithm and compared with the model obtained by the traditional standard two-stage approach. Individual pharmacokinetic parameter estimates were calculated by weighted nonlinear least-squares regression and by maximum a posteriori probability Bayesian estimator. Individual parameter estimates obtained with four D-optimally timed serum samples (OSS4) showed excellent correlation with parameter estimates obtained by using full data sets. The parameters of interest, clearance and volume of distribution, showed excellent agreement (R2 = 0.89 and R2 = 0.86). The ceftazidime population models were described as two-compartment kslope models, relating elimination constants to renal function. The NPEM-OSS4 model was described by the equations kel = 0.06516+ (0.00708.CLCR) and V1 = 0.1773 +/- 0.0406 liter/kg where CLCR is creatinine clearance in milliliters per minute per 1.73 m2, V1 is the volume of distribution of the central compartment, and kel is the elimination rate constant. Predictive performance evaluation for 31 patients with data which were not part of the model data sets showed that the NPEM-ALL model performed best, with significantly better precision than that of the standard two-stage model (P < 0.001). Predictions with the NPEM-OSS4 model were as precise as those with the NPEM-ALL model but slightly biased (-2.2 mg/liter; P < 0.01). D-optimal monitoring strategies coupled with population modeling results in useful and cost-effective population models and will be of advantage in clinical practice, as it allows pharmacokinetic-pharmacodynamic modeling with sparse data, thus describing the relationship between ceftazidime exposure and response in the treatment of acute exacerbations in patients with cystic fibrosis. PMID- 8723445 TI - Chromosomal gentamicin resistance transposon Tn3706 in Streptococcus agalactiae B128. AB - Streptococcus agalactiae B128 is the only highly gentamicin-resistant group B streptococcal (GBS) strain described so far. This strain carries a chromosomal gentamicin resistance transposon, designated Tn3706, which is similar, if not identical, to the Tn4001 and Tn5281 transpons detected in Staphylococcus aureus and Enterococcus faecalis, respectively. Transposition of Tn3706 occurred onto the GBS plasmid pIP501 in two different loci of its 7.5-kb AvaII fragment carrying the genes for chloramphenicol and erythromycin resistance. Molecular analysis of pIP501 derivatives showed that Tn3706 is composed of a central fragment containing the aac6'-aph2" gene; this fragment is flanked by two tandemly repeated copies of IS256 at the 5' extremity of the resistance gene and a single inverted copy of IS256 at its 3' extremity. The two tandemly repeated copies of IS256 were separated by a 6-bp segment identical to that found, in the same orientation, in the IS256-aac6'-aph2" junction. The hybrid replicons pIP501::Tn3706 were found to be structurally unstable following conjugative transfer between GBS strains. Numerous individual copies of IS256 were detected in B128, but this insertion sequence was not found in the 11 wild-type, gentamicin-susceptible GBS strains studied. PMID- 8723447 TI - Lack of effect of vancomycin and gentamicin on auditory function in guinea pigs. AB - The dispositions of vancomycin (VCM) and gentamicin (GM) in plasma and perilymph after single and multiple administrations and the effects of multiple administrations of VCM or GM alone and the combination of both drugs on auditory function were studied in male guinea pigs. The pharmacokinetic parameters of VCM and GM obtained from plasma drug concentration-time data after single and multiple (22 days) intramuscular administrations of VCM (200 mg/kg of body weight) alone and GM (50 mg/kg) alone were not significantly different from those after combined administration of VCM (200 mg/kg) and GM (50 mg/kg). There was no change in the penetration ratio of VCM and GM into perilymph between administration of VCM or GM alone and the combination of both drugs. Furthermore, the hearing threshold of guinea pigs was not affected by VCM or GM alone or the combination of both drugs within the range of therapeutic VCM and GM levels in plasma in humans. PMID- 8723448 TI - Concentrations of doxycycline and penicillin G in sera and cerebrospinal fluid of patients treated for neuroborreliosis. AB - Concentrations of doxycycline and penicillin G in serum and cerebrospinal fluid (CSF) were analyzed in 46 patients during treatment for neuroborreliosis. Twenty patients were treated intravenously with penicillin G at 3 g every 6 h (q6h), and 26 patients were treated orally with doxycycline at 200 mg q24h. All samples were collected on day 13 of treatment. The median concentrations of penicillin G in serum were 0.5, 37, and 5.6 micrograms/ml before and 1 and 3 h after drug administration, and that in CSF was 0.5 (range, 0.3 to 1.6) microgram/ml after 2 to 3 h. The median concentrations of doxycycline in serum were 2.1, 6.1, and 4.7 micrograms/ml before and 2 and 6 h after drug administration, and that in CSF was 0.6 (range, 0.4 to 2.5) microgram/ml after 4 h. All patients had concentrations of penicillin G or doxycycline in CSF above the lowest reported MICs of penicillin G (0.003 microgram/ml) and doxycycline (0.12 microgram/ml) for Borrelia burgdorferi. However, no patients had a drug concentration in CSF above the highest reported MIC of penicillin G (8 micrograms/ml), and only one had a drug concentration in CSF above the highest reported MIC of doxycycline (2 micrograms/ml), despite good clinical response to treatment. No treatment failure or relapse was observed during a 1-year follow-up, although one patient treated with penicillin G and one treated with doxycycline were retreated because of residual pain. The chosen dosages of penicillin G and doxycycline seem to give sufficient concentrations in serum and CSF for the treatment of neuroborreliosis. PMID- 8723449 TI - Monotherapy with meropenem versus combination therapy with ceftazidime plus amikacin as empiric therapy for fever in granulocytopenic patients with cancer. The International Antimicrobial Therapy Cooperative Group of the European Organization for Research and Treatment of Cancer and the Gruppo Italiano Malattie Ematologiche Maligne dell'Adulto Infection Program. AB - Combinations of beta-lactams plus aminoglycosides have been standard therapy for suspected infections in granulocytopenic cancer patients, especially those with profound long-lasting granulocytopenia. With the advent of new broad-spectrum bactericidal antibiotics such as extended-spectrum cephalosporins or carbapenems, the need to combine beta-lactams with aminoglycosides became more controversial. The objective of this prospective randomized multicenter study was to compare the efficacy, safety, and tolerance of meropenem monotherapy with those of the combination of ceftazidime plus amikacin for the empirical treatment of fever in granulocytopenic cancer patients. Of 1,034 randomized patients, 958 were assessable in the intent-to-treat analysis for response to antibacterial therapy, including 483 in the meropenem group and 475 in the ceftazidime-plus-amikacin group. The median durations of neutropenia were 16 and 17 days, respectively. A successful outcome was reported in 270 of 483 (56%) patients treated with monotherapy compared with 245 of 475 (52%) patients treated with the combination group (P = 0.20). The success rates in the monotherapy group and the combination group were similar by type of infection (single gram-negative bacteremia, single gram-positive bacteremia, clinically documented infection, and possible infection). The occurrence of further infections assessed in patients for whom the allocated regimen was not modified did not differ between the two groups (12% in both groups). Mortality due to the presenting infection or further infection was relatively low (8 patients treated with the monotherapy compared with 13 patients treated with the combination). A total of 1,027 patients were evaluable for adverse events; the proportion of those who developed adverse effects was similar between the two groups (29% in both groups), and only 19 (4%) patients in the monotherapy group and 31 (6%) in the combination group experienced an adverse event related or probably related to the study drug. Allergic reactions were the only reason for stopping the protocol antibiotic(s) (3 and 5 patients, respectively). This study confirms that monotherapy with meropenem is as effective as the combination of ceftazidime plus amikacin for the empiric treatment of fever in persistently granulocytopenic cancer patients, and both regimens were well tolerated. PMID- 8723450 TI - Possible reason for preferential damage to renal tubular epithelial cells evoked by amphotericin B. AB - An important determinant of nephrotoxicity, which is the major complication of long-term amphotericin B treatment, is dysfunction of distal tubular epithelial cells. The underlying cause for this rather selective damage to the cells is unknown. In the present investigation, it was shown that kidney epithelial cells were initially damaged by amphotericin B at concentrations of 2.5 to 10 micrograms/ml, as demonstrable by a dramatic drop in cellular K+ levels. Cells could recover from the initial toxic action of the polyene if they were kept in medium of neutral pH, and cellular K+ levels returned to normal after 6 h. However, the recovery mechanisms failed at lower pHs of 5.6 to 6.0. At low pHs, cells became progressively depleted of ATP; they leaked lactate dehydrogenase and became irreversibly damaged after approximately 6 h. The possibility that the low pH characteristic of the distal tubulus lumen renders the renal epithelial cells particularly vulnerable to the toxic action of amphotericin B is raised. The concept is in line with an earlier report that alkalization ameliorates amphotericin B nephrotoxicity in rats. PMID- 8723451 TI - In vitro effect of tinidazole and furazolidone on metronidazole-resistant Trichomonas vaginalis. AB - Trichomonas vaginalis is a common sexually transmitted protozoan parasite. Although often considered simply a nuisance infection, T. vaginalis has been implicated in premature rupture of placental membranes and increases in the risk of acquiring human immunodeficiency virus. Metronidazole, a 5-nitroimidazole, is currently the drug of choice to treat T. vaginalis infection. Because some patients have severe reactions to metronidazole and others are infected with metronidazole-resistant T. vaginalis, we were prompted to investigate alternative therapies. Tinidazole, another 5-nitroimidazole used in other countries to treat T. vaginalis infections, and furazolidone, a nitrofuran presently used to treat giardiasis and infections with some anaerobic enteric bacteria, were investigated for effectiveness against 9 metronidazole-susceptible and 12 metronidazole resistant T. vaginalis patient isolates. The in vitro aerobic and anaerobic minimum lethal concentrations (MLC) and the time for drug efficacy were determined. Tinidazole killed the metronidazole-susceptible isolates at a low MLC but was effective against only 4 of the 12 metronidazole-resistant isolates. In contrast, furazolidone was effective at a low MLC for all isolates. When tinidazole was effective, it required > 6 h to kill trichomonads. However, furazolidone killed both metronidazole-susceptible and resistant trichomonads within 2 to 3 h of exposure. These data suggest that furazolidone may be a good candidate for treating metronidazole-resistant trichomoniasis and that further investigation of this drug is warranted. PMID- 8723452 TI - Cloning and nucleotide sequence of the DNA gyrase gyrA gene from the fish pathogen Aeromonas salmonicida. AB - The DNA gyrase gyrA gene from the fish pathogen Aeromonas salmonicida 2148/89 was cloned, and the nucleotide sequence was determined. An open reading frame of 2,766 nucleotides was identified and was found to encode a protein of 922 amino acids with a calculated molecular mass of 101.1 kDa. The derived amino acid sequence shared a high degree of identity with other DNA gyrase A proteins, in particular, with other gram-negative GyrA sequences. When the amino acid sequence of A. salmonicida GyrA was compared with that of Escherichia coli GyrA, a number of conserved residues were present at identical coordinates, including the catalytic Tyr residue at position 122 (Tyr-122) and residues whose substitution confers quinolone resistance, notably, Ser-83, Ala-67, Gly-81, Asp-87, Ala-84, and Gln-106. An intragenic region corresponding to 48 amino acids, which is not present in E. coli or other bacteria, was identified in the C-terminal part of A. salmonicida GyrA. This intragenic region shared sequence identity with various DNA-binding proteins of both prokaryotic and eukaryotic origins. PMID- 8723453 TI - Protein binding of clindamycin in sera of patients with AIDS. AB - Patients with AIDS have altered pharmacokinetics of clindamycin compared with those of healthy control subjects. In an attempt to better understand these differences, we undertook a study of protein binding of clindamycin in sera of patients with AIDS. Fifteen patients with AIDS and 15 healthy volunteers were given a single 600-mg dose of clindamycin orally and intravenously, and serum samples were collected at three time points corresponding to high, midpoint, and low clindamycin concentrations. Protein binding was determined by ultrafiltration, and total and unbound clindamycin concentrations were measured with a gas chromatography assay. AIDS patients had alpha 1-acid glycoprotein values approximately twice those of healthy volunteers (mean +/- standard deviation, 103 +/- 27 versus 61 +/- 11 mg/dl; P = 0.001). Overall, serum protein binding levels were higher in AIDS patients (mean +/- standard deviation, 83 +/- 7 versus 78% +/- 8%; P = 0.0001), which is likely the result of increased alpha 1 acid glycoprotein levels in these patients. Total concentrations of clindamycin in plasma were significantly higher in AIDS patients at most time points studied, while unbound serum clindamycin concentrations did not differ among the groups at each sampling time after both oral and intravenous dosing. Increased protein binding may partly explain the altered pharmacokinetic disposition of clindamycin in AIDS patients; however, other factors cannot be excluded. PMID- 8723454 TI - Pharmacokinetics of vancomycin in critically ill infants undergoing extracorporeal membrane oxygenation. AB - Extracorporeal membrane oxygenation (ECMO) is a widely used therapy for neonates with respiratory failure. Because of sepsis, many of these infants require antibiotics like vancomycin during ECMO treatment. ECMO transiently alters renal function and increases the circulating blood volume by 75%. Initial vancomycin pharmacokinetics were determined in 12 infants undergoing ECMO to determine an adequate drug administration regimen. Vancomycin dosage was based on current recommendations for weight and gestational age. Pharmacokinetic parameters were determined by fitting the data to a two compartment model. This study yielded a mean steady-state volume of distribution of 1.1 +/- 0.5 (range, 0.6 to 2.1) liters/kg and a mean vancomycin clearance of 0.78 +/- 0.19 (range, 0.49 to 1.07) ml/min/kg. The mean vancomycin half-life was 16.9 +/- 9.5 (range, 8.8 to 42.9) h. Nomogram-calculated creatinine clearance was a significant predictor of vancomycin terminal rate constant and clearance. These data suggest alterations in the pharmacokinetics of vancomycin in infants on ECMO. With the goal of achieving vancomycin concentrations in serum above the MIC for the offending pathogen while using the least amount of the drug necessary, new administration guidelines for term infants without renal impairment undergoing ECMO should be 20 mg of vancomycin per kg at an interval of 24 h. With significant renal impairment, the interval should be extended on the basis of concentrations in serum. In comparison with previously published data, the neonates undergoing ECMO in our study demonstrated a much larger volume of distribution, a lower clearance, and consequently a longer vancomycin half-life. PMID- 8723455 TI - Modeling combinations of antiretroviral agents in vitro with integration of pharmacokinetics: guidance in regimen choice for clinical trial evaluation. AB - We propose a method for the selection of doses and dosing schedule for drugs to be used in combination. This approach uses the simulation of steady-state concentrations of the drugs in the combination and overlays these concentrations onto a three-dimensional effect surface. The MacSynergy II program is used to construct the three-dimensional drug interaction surface from the direct evaluation of drug combination effect in vitro. The study examined the combination of an inhibitor of the human immunodeficiency virus protease, A 77003, and the nucleoside analog zidovudine. Zidovudine concentrations from a steady-state interval were simulated on the basis of the administration of 100 mg every 12 h by mouth, while for A-77003 simulation profiles were for intravenous administration of 800 mg every 4 h as well as a continuous infusion of 200 mg/h. The average percentage of the maximal effect was taken as a measure of regimen effectiveness. Three different schedules of administration were examined. If both drugs were to be administered simultaneously, the model predicts a mean maximal effect of a steady-state interval (12 h) of 67%. If the drug doses were offset by 2 h, the mean maximal effect predicted was 71%. If A-77003 was to be given by continuous infusion, the mean maximal effect predicted was 90%. This method holds promise as a way of quickly evaluating potential combinations of agents that takes into account the drug interaction in a mathematically robust way and that allows the evaluation of the effect of each drug's pharmacokinetic profile. PMID- 8723456 TI - Bactericidal activities of cefprozil, penicillin, cefaclor, cefixime, and loracarbef against penicillin-susceptible and -resistant Streptococcus pneumoniae in an in vitro pharmacodynamic infection model. AB - We examined the bactericidal activities of penicillin, cefprozil, cefixime, cefaclor, and loracarbef against three clinical isolates of Streptococcus pneumoniae which were susceptible, moderately susceptible, and resistant to penicillin. An in vitro two-compartment glass infection model was used to simulate human pharmacokinetics in the presence of bacteria. Also, changes in organism susceptibility and development of resistant subpopulations were evaluated. Simulated pediatric dosage regimens and target peak concentrations in the central compartment were as follows: penicillin V-potassium, 26 mg/kg of body weight every 6 h (q6h) and 14 micrograms/ml; cefaclor, 13.4 mg/kg q8h and 16 micrograms/ml; loracarbef, 15 mg/kg q12h and 19 micrograms/ml; cefprozil, 15 mg/kg q12h and 11 micrograms/ml; and cefixime, 8mg/kg q24h and 4 micrograms/ml. Targeted half-lives of each agent were 1 h for penicillin, cefaclor, and loracarbef; 1.3 h for cefprozil; and 3.5 h for cefixime. Growth controls were performed at two different pump rates, 0.8 and 2.0 ml/min (half-lives = 3.5 and 1 h, respectively). Each isolate demonstrated autolysis at the lower rate which was attributed to a decreased supply of fresh nutrients available to the organisms in the infection compartment. Against the susceptible isolate, the time to 99.9% killing was statistically significant between penicillin V-potassium and both cefaclor and cefixime (P < 0.029). Loracarbef never achieved a 99.9% reduction in the inoculum. At 48 h penicillin, cefprozil, and cefaclor were equivalent in extent of killing. Against the intermediately resistant isolate, cefprozil was superior to all other regimens with respect to rate of killing (P < 0.013) and extent of killing at 24 h (P < 0.0003). At 48 h penicillin, cefprozil, and cefaclor were equivalent in extent of killing. All of the regimens exhibited inferior activity against this penicillin-resistant isolate. A 99.9% kill was never obtained with any of the regimens, nor was there an appreciable decrease in the colony counts. In conclusion, it appears that cefprozil, penicillin, and cefaclor are effective therapies against sensitive and even intermediately sensitive isolates of S. pneumoniae. However, none of the oral therapies appear to be of any benefit against penicillin-resistant isolates. The in vitro model may be an effective tool in evaluating other multiple-dose therapies against this fastidious organism, since the continual supply of fresh medium maintains the viability of S. pneumoniae with minimal stationary-phase autolysis. PMID- 8723457 TI - Ciprofloxacin pharmacokinetics in burn patients. AB - Many drugs exhibit altered pharmacokinetic parameters in burn patients. We prospectively evaluated the pharmacokinetics of ciprofloxacin in eight burn patients with active infections. Each patient received a 400-mg dose of ciprofloxacin intravenously (i.v.) every 8 h, with each dose infused over 1 h by using a rate control device. Blood samples for analysis of plasma ciprofloxacin concentrations, determined by high-performance liquid chromatography, were obtained immediately predose, at the end of the infusion, and 1, 2, 3, 4, 5, 6, and 7 h after the end of the infusion. Urine was collected from 0 to 2, 2 to 4, and 4 to 8 h following the same dose, and an aliquot was saved for determination of the ciprofloxacin concentration. Urine was also collected for 24 h prior to this dose for measurement of creatinine clearance (CLCR). Pharmacokinetic parameters were estimated by noncompartmental analysis. Mean maximum and minimum plasma ciprofloxacin concentrations were 4.2 +/- 1.1 and 0.70 +/- 0.55 microgram/ml, respectively. Mean values for clearance (CL), renal clearance (CLR), volume of distribution, terminal elimination rate constant, half-life (t1/2), and area under the concentration-time curve (AUC) were 29.1 +/- 17.5 liters/h, 13.5 +/- 10.1 liters/h, 1.75 +/- 0.41 liters/kg, 0.222 +/- 0.098 h-1, 4.5 +/- 3.9 h, and 20.7 +/- 16.6 micrograms.h/ml, respectively. CL was higher and t1/2 was shorter than noted in previous studies of acutely ill, hospitalized patients. A good correlation was noted between creatinine clearance CL(CR) and both total ciprofloxacin CL (r = 0.85) and CLR (r = 0.84). A moderate inverse correlation was noted between percent body surface area burned and total ciprofloxacin CL (r = -0.55). An AUC/MIC ratio above 125 SIT-1 (where SIT is serum inhibitory titer), which has been strongly correlated with clinical response and time to bacterial eradication, was achieved in five of eight patients (63%) with a MIC of 0.25 microgram/ml. At a ciprofloxacin dosage of 400 mg i.v. every 12 h, an AUC/MIC ratio above 125 SIT-1 would have been achieved in only two of eight patients (25%). We conclude that ciprofloxacin CL is highly variable, but generally increased, in burn patients compared with that in acutely ill, general medical and surgical patients. Because of an increase in CL, a ciprofloxacin dosage of 400 mg i.v. every 8 h is more likely to produce the desired response in burn patients than the same dose given every 12 h. PMID- 8723458 TI - Alterations in the DNA topoisomerase IV grlA gene responsible for quinolone resistance in Staphylococcus aureus. AB - A 4.2-kb DNA fragment conferring quinolone resistance was cloned from a quinolone resistant clinical isolate of Staphylococcus aureus and was shown to possess a part of the grlB gene and a mutated grlA gene. S-80-->F and E-84-->K mutations in the grlA gene product were responsible for the quinolone resistance. The mutated grlA genes responsible for quinolone resistance were dominant over the wild-type allele, irrespective of gene dosage in a transformation experiment with the grlA gene alone. However, dominance by mutated grlA genes depended on gene dosage when bacteria were transformed with the grlA and grlB genes in combination. Quinolone resistant gyrA mutants were easily isolated from a strain, S. aureus RN4220, carrying a plasmid with the mutated grlA gene, though this was not the case for other S. aureus strains lacking the plasmid. The elimination of this plasmid from such quinolone-resistant gyrA mutants resulted in marked increases in quinolone susceptibility. These results suggest that both DNA gyrase and DNA topoisomerase IV may be targets of quinolones and that the quinolone susceptibility of organisms may be determined by which of these enzymes is most quinolone sensitive. PMID- 8723459 TI - In vivo antibacterial effects of simulated human serum profiles of once-daily versus thrice-daily dosing of amikacin in a Serratia marcescens endocarditis experimental model. AB - Once-daily dosage of aminoglycosides is currently under consideration. The lower toxicity of this regimen has been clearly established, but there are conflicting experimental and clinical data concerning its efficacy. It is inadvisable to optimize human therapy by extrapolation from experimental studies since animal and human pharmacokinetics differ. The simulation of human pharmacokinetics in experimental infectious models would seem to offer a more rational approach. We used computer-controlled infusion of amikacin at a variable flow rate to simulate human pharmacokinetics in a Serratia marcescens rabbit endocarditis model and to compare two therapeutic regimens (once-daily versus thrice-daily doses). The doses corresponded to simulations of 15 and 30 mg/kg of body weight per day in humans, and antibacterial activity was measured in vegetations (Veg) after 24 h of treatment. The results show that the dose corresponding to 15 mg/kg/day failed to produce a significant reduction of CFU (6.8 +/- 0.9 and 6.4 +/- 0.8 log10 CFU/g of Veg, respectively, for once-daily and thrice-daily doses versus 7.6 +/- 1.0 for controls). A significant reduction was observed only for the dose corresponding to 30 mg/kg/day in humans (5.2 +/- 1.5 and 5.4 +/- 1.1 log10 CFU/g of Veg, respectively, for the two regimens). With this model, the efficacy of amikacin was similar for both regimens after 24 h of treatment simulating human pharmacokinetics. PMID- 8723460 TI - Comparison of the beta-lactamase gene cluster in clonally distinct strains of Enterococcus faecalis. AB - Ten beta-lactamase-producing Enterococcus faecalis isolates were examined for the presence of the staphylococcal beta-lactamase repressor and antirepressor genes. Four isolates, previously shown to be unrelated to each other by pulsed-field gel electrophoresis analysis, were positive for both genes by PCR, although beta lactamase production was not induced with methicillin. Six isolates, previously shown to be clonally related, were negative for both genes by PCR. The blaZ sequences of eight beta-lactamase-producing E. faecalis isolates were determined. Seven isolates from five distinct clones had sequences identical to that previously reported for E. faecalis HH22, regardless of whether the repressor or antirepressor was demonstrated by PCR. However, blaZ from one isolate differed from those of the other enterococci by 11 nucleotides; this isolate is part of the large clone, as defined by pulsed-field gel electrophoresis and multilocus enzyme analysis, that includes HH22. These findings suggest either that enterococci have acquired the bla gene cluster from more than one source or that the gene cluster has undergone considerable change since acquisition by this clone. PMID- 8723461 TI - Ofloxacin versus standard therapy in treatment of community-acquired pneumonia requiring hospitalization. Pneumonia Study Group. AB - Community-acquired pneumonia occurs 3 to 4 million times per year in the United States, accounting for about 500,000 hospitalizations annually. Empiric treatment is usually instituted because of a lack of early organism-specific diagnostic tests. This study compared empiric therapy with ofloxacin to standard antibiotic regimens (usually a beta-lactam with or without a macrolide) for patients hospitalized for community-acquired pneumonia. Therapy was administered to 298 patients (146 receiving ofloxacin and 152 receiving standard therapy); 227 patients (ofloxacin, 109; standard treatment, 118) were evaluable for treatment efficacy. The most common pyogenic respiratory pathogens were Haemophilus influenzae (30 isolates) and Streptococcus pneumoniae (24 isolates). There was evidence of infection with either Mycoplasma pneumoniae (38 patients), Chlamydia pneumoniae (40 patients), or a Legionella sp. (8 patients) in a total of 79 patients (35%). The clinical success rates were similar in both groups among evaluable patients (92%, ofloxacin; 87%, standard therapy) and among patients with atypical respiratory pathogens (88%, ofloxacin; 81%, standard therapy). The mean numbers (+/- the standard deviations) of intravenous doses of antibiotics were 7.5 +/- 8.0 in the ofloxacin group and 18.4 +/- 18.5 in the standard therapy group (P < 0.001); the mean number of oral doses of ofloxacin per patient was 19.7 +/- 11.2, compared with 30.2 +/- 16.0 oral antibiotic doses in the standard therapy group (P < 0.001). All treatments were well tolerated and associated with no significant clinical or laboratory abnormalities. The findings of this study indicate that ofloxacin is active against traditional bacterial pathogens as well as the major atypical respiratory pathogens. When given as monotherapy for the empiric treatment of community-acquired pneumonia, ofloxacin is as effective as standard antimicrobial therapy. PMID- 8723462 TI - Interaction between ganciclovir and foscarnet as inhibitors of duck hepatitis B virus replication in vitro. AB - Safe and effective treatments for chronic hepatitis B virus (HBV) infection have yet to be developed. Both ganciclovir (9-[1,3-dihydroxy-2-propoxymethyl]guanine) and foscarnet (trisodium phosphonoformate hexahydrate) are potent inhibitors of hepadnavirus replication when used individually in vitro and in vivo. However, the clinical usefulness of each drug is reduced by dose-limiting toxicity, especially during long-term monotherapy. Here we demonstrate additive inhibition of duck HBV DNA replication in cultures of primary duck hepatocytes congenitally infected with duck HBV by combinations of ganciclovir and foscarnet at low, clinically achievable concentrations. These results suggest that the effects of ganciclovir and foscarnet against HBV may be additive in vivo. PMID- 8723463 TI - Characterization of streptomycin resistance mechanisms among Mycobacterium tuberculosis isolates from patients in New York City. AB - From a collection of 367 isolates of Mycobacterium tuberculosis from patients in New York City in 1994, 45 isolates (12.3%) were resistant in vitro to 2 micrograms or more of streptomycin (SM) per ml. We further evaluated these isolates for levels of SM resistance and for mutations previously associated with resistance in the rpsL (S12 ribosomal protein) gene and the rrs (16S rRNA)-coding region. Twenty-four isolates, representing nine distinct patterns of susceptibility to antituberculosis drugs, were resistant to 500 micrograms of SM per ml and shared a common point mutation at nucleotide 128 in the rpsL gene. This mutation, which substitutes lysine for arginine in the S12 ribosomal binding protein, was not present in isolates with low-level SM resistance or in SM susceptible control isolates. Among 20 isolates with low-level SM resistance, one possessed a substitution (C-->G865) in the 912 loop of the rrs gene. No mutations in the 530 loop of the rrs coding region were detected, suggesting the presence of an alternative SM resistance mechanism in 19 isolates. Single-strand conformation polymorphisms of mutants were readily detected by a nonradioactive gel screen. PMID- 8723464 TI - A novel antiviral agent which inhibits the endonuclease of influenza viruses. AB - A novel anti-influenza virus compound, flutimide, was identified in extracts of a recently identified fungal species, Delitschia confertaspora (F. Pelaez, J.D. Polishook, M. Valldosera, and J.Guarro, Mycotaxon 50:115-122, 1994). The compound, a substituted 2,6-diketopiperazine, selectively inhibited the cap dependent transcriptase of influenza A and B viruses and had no effect on the activities of other polymerases. Similar to the 4-substituted 2,4-dioxobutanoic acids, a series of transcriptase inhibitors which we described previously (J. Tomassini, H. Selnick, M.E. Davies, M.E. Armstrong, J. Baldwin, M. Bourgeois, J.Hastings, D. Hazuda, J. Lewis, W. McClements, G. Ponticello, E. Radzilowski, G. Smith, A. Tebben, and A. Wolfe, Antimicrob. Agents Chemother. 38:2827-2837, 1994), this inhibitor, which is a natural product, affected neither the initiation nor the elongation of influenza virus mRNA synthesis, but it specifically targeted the cap-dependent endonuclease of the transcriptase. Additionally, the compound was inhibitory to the replication of influenza A and B viruses in cell culture. The selective antiviral properties of this compound further demonstrate the utility of influenza virus endonuclease as a target of antiviral agents. PMID- 8723466 TI - Penetration and bactericidal activity of cefixime in synovial fluid. AB - The penetration of oral cefixime into the synovial fluids of 16 patients (mean age, 50.6 years) who underwent joint taps for rheumatic noninfectious disorders was examined. The patients were each given a single dose (400 mg) 2 to 24 h prior to the tap. Cefixime concentrations in serum and joint fluid samples were measured by high-performance liquid chromatography, and the bactericidal activities of these fluids against three isolates each of Haemophilus influenzae and Escherichia coli were examined. The highest concentrations in serum and synovial fluid were achieved 4 h following drug intake, the mean values being 2.8 and 2.03 micrograms/ml, respectively. Effective bactericidal activities (bactericidal titer, > 1:2) against E. coli and H. influenzae were demonstrated in serum and joint fluid up to 10 h following oral intake of cefixime. These results suggest that cefixime penetrates well into joint fluid, achieving levels above the MIC for E. coli lasting as long as 10 h and levels above the MIC for H. influenzae lasting up to 24 h after administration. Good bactericidal activity against susceptible bacterial isolates was observed for at least 10 h after dosing. PMID- 8723465 TI - Effects of pentoxifylline or dexamethasone in combination with amphotericin B in experimental murine cerebral cryptococcosis: evidence of neuroexcitatory pathogenic mechanisms. AB - In a murine model of intracerebral infection by Cryptococcus neoformans the therapeutic effects of pentoxifylline or dexamethasone were studied alone and in combination with amphotericin B. Assessed parameters were mean survival time, brain histopathology index, amounts of glutamate and gamma-aminobutyric acid in the brain, and yeast CFU per brain. Survival increased significantly in mice treated with dexamethasone, amphotericin B, amphotericin B plus dexamethasone, and amphotericin B plus pentoxifylline; the latter had significantly longer survival than other treated groups. Indices of histopathological damage were similar in all treated groups. In infected untreated mice, the amounts of glutamate in the brain were decreased, presumably by depletion. In mice treated with amphotericin B plus dexamethasone, glutamate levels returned to the range of control mice. No differences in the amounts of gamma-aminobutyric acid were found between control and treatment groups. Brain fungal counts were significantly lower in mice treated with amphotericin B, amphotericin B plus dexamethasone, and amphotericin B plus pentoxifylline than in untreated animals. In this model, pentoxifylline in combination with amphotericin B improved survival, decreasing the fungal burden, and has potential as adjuvant therapy in cerebral cryptococcosis. PMID- 8723467 TI - In vitro and in vivo antibacterial activities of CS-940, a new 6-fluoro-8 difluoromethoxy quinolone. AB - The in vitro and in vivo activities of CS-940, a new 6-fluoro-8-difluoromethoxy quinolone, were compared with those of ciprofloxacin, tosufloxacin, sparfloxacin, and levofloxacin. The in vitro activity of CS-940 against gram-positive bacteria was nearly equal to or greater than those of the other quinolones tested. In particular, CS-940 was two to eight times more active against methicillin resistant Staphylococcus aureus than the other quinolones, at the MIC at which 90% of the clinical isolates are inhibited. Against gram-negative bacteria, the activity of CS-940 was comparable to or greater than those of tosufloxacin, sparfloxacin, and levofloxacin, while it was lower than that of ciprofloxacin. The activity of CS-940 was largely unaffected by medium, inoculum size, or the addition of horse serum, but it was decreased under acidic conditions, as was also seen with the other quinolones tested. CS-940 showed potent bactericidal activity against S. aureus, Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa. In oral treatment of mouse systemic infections caused by S. aureus, Streptococcus pneumoniae, Streptococcus pyogenes, E. coli, K. pneumoniae, Serratia marcescens, and P. aeruginosa, CS-940 was more effective than ciprofloxacin, sparfloxacin, and levofloxacin against all strains tested. Against experimental pneumonia with K. pneumoniae in mice, CS-940 was the most effective of all the quinolones tested. These results suggest that CS-940 may be effective in the therapy of various bacterial infections. PMID- 8723468 TI - Antimicrobial resistance of Streptococcus pneumoniae recovered from outpatients in the United States during the winter months of 1994 to 1995: results of a 30 center national surveillance study. AB - A total of 1,527 clinically significant outpatient isolates of Streptococcus pneumoniae were prospectively collected in 30 different U.S. medical centers between November 1994 and April 1995. Overall, 23.6% of strains were not susceptible to penicillin, with 14.1% intermediate and 9.5% high-level resistant. The frequencies of recovery of intermediate and high-level resistant strains varied considerably between different medical centers and in different geographic areas. In general, intermediate and high-level penicillin resistance was most common with isolates of S. pneumoniae recovered from pediatric patients. The in vitro activities of 22 other antimicrobial agents were assessed against this collection of isolates. Ampicillin was consistently 1 twofold dilution less active than penicillin. Amoxicillin and amoxicillin-clavulanate were essentially equivalent to penicillin in activity. The rank order of activity for cephalosporins was cefotaxime = ceftriaxone > or = cefpodoxime > or = cefuroxime > cefprozil > or = cefixime > cefaclor = loracarbef > cefadroxil = cephalexin. The National Committee for Clinical Laboratory Standards [Performance Standards for Antimicrobial Susceptibility Testing, Sixth Information Supplement (M100-S6), 1995] has established MIC breakpoints for resistance (i.e., > or = 2 micrograms/ml) with three cephalosporins versus S. pneumoniae, namely, cefotaxime, ceftriaxone, and cefuroxime. The overall percentages of strains resistant to these three antimicrobial agents were 3, 5, and 12, respectively. The overall frequency of resistance was 10% with all three macrolides examined in this study, clarithromycin, erythromycin, and azithromycin. The overall percentages of chloramphenicol, tetracycline, and trimethoprim-sulfamethoxazole resistance were 4.3, 7.5, and 18, respectively. The resistance percentages among the cephalosporins, macrolides, chloramphenicol, tetracycline, and trimethoprim sulfamethoxazole were consistently higher among penicillin-intermediate strains than among susceptible isolates and even higher still among organisms expressing high-level penicillin resistance. Multiply resistant strains represented 9.1% of the organisms examined in this study. Finally, rifampin resistance was uncommon (i.e., 0.5%), and vancomycin resistance was not detected. The quinopristin dalfopristin combination was consistently active at concentrations of 0.25 to 4 micrograms/ml, but rates of resistance could not be determined in the absence of established interpretive criteria for MIC results. PMID- 8723469 TI - Therapy of visceral leishmaniasis due to Leishmania infantum: experimental assessment of efficacy of AmBisome. AB - The tolerance and efficacy of amphotericin B (AmB) deoxycholate (Fungizone) were compared with those of liposomal AmB (AmBisome) in a murine model of visceral leishmaniasis induced by Leishmania infantum. Control groups consisted of untreated mice and mice treated with a pentavalent antimonial (Glucantime). BALB/c mice were infected intravenously on day 0 with 10(7) promastigotes of L. infantum and then treated from day 7 to 17 (early treatment group) or from day 60 to 70 (delayed treatment group). The pentavalent antimonial was administered daily by intraperitoneal injection, whereas AmB formulations were administered intravenously on alternate days. On days 20, 60, and 120 (early treatment group) and on days 72 and 125 (delayed treatment group), parasite burdens in the liver, spleen, and lungs were determined by subculturings using a microtitration method. A dose range study showed that administration of AmBisome at the well-tolerated doses of 5 or 50 mg/kg of body weight completely eradicated the parasites from the tissues. At 0.8 mg/kg, AmBisome proved more efficacious than AmB deoxycholate administered at the same dose. We also compared the levels of AmB deoxycholate and AmBisome in plasma and tissue. Mice treated with AmBisome had levels of AmB in tissue much higher than did AmB deoxycholate-treated mice with persistent detectable levels 14 weeks after treatment. These results seem to account for the remarkable efficacy of the liposomal formulation of AmB in the treatment of visceral leishmaniasis due to L. infantum. PMID- 8723470 TI - Importance of penicillinase production for activity of penicillin alone or in combination with sulbactam in experimental endocarditis due to methicillin resistant Staphylococcus aureus. AB - The activity of penicillin, alone and in combination with sulbactam, against a heterogeneously methicillin-resistant, penicillinase-producing clinical isolate of Staphylococcus aureus and its penicillinase-negative derivative was investigated in vitro and in a rabbit experimental endocarditis model. Penicillin was significantly more effective than vancomycin against the penicillinase negative derivative in vivo (P < 0.001), and it sterilized 25% of the vegetations. The combination of penicillin and sulbactam exhibited an in vivo synergistic effect on the penicillinase-producing strain (P < 0.01) but did not produce any advantage over treatment with vancomycin, even when a high dose of sulbactam was used (100 mg/kg of body weight every 6 h). This combination was significantly less effective against the penicillinase-producing strain than was penicillin alone against the penicillinase-negative derivative (P < 0.03). In addition, the most resistant subpopulation of the surviving bacteria, which grew on agar containing 16 micrograms of methicillin per ml, was detected in 5 of 6 animals treated with penicillin and a high dose of sulbactam against the penicillinase-producing strain compared with only 1 of 12 animals treated with penicillin alone against the penicillinase-negative derivative (P < 0.01). We conclude that penicillin is highly effective against penicillinase-negative methicillin-resistant S. aureus and that penicillinase production, rather than methicillin resistance, appears to be the limiting factor for the activity of the penicillin-sulbactam combination against penicillinase-producing, methicillin resistant S. aureus. PMID- 8723471 TI - Effect of recombinant human gamma interferon on intracellular activities of antibiotics against Listeria monocytogenes in the human macrophage cell line THP 1. AB - Listeria monocytogenes is a facultative intracellular pathogen which enters cells by endocytosis and reaches phagolysosomes from where it escapes and multiplies in the cytosol of untreated cells. Exposure of macrophages to gamma interferon (IFN gamma) restricts L. monocytogenes to phagosomes and prevents its intracellular multiplication. We have tested whether IFN-gamma also modulates the susceptibility of L. monocytogenes to antibiotics. We selected drugs from three different classes displaying marked properties concerning their cellular accumulation and subcellular distribution, namely, ampicillin (not accumulated by cells but present in cytosol), azithromycin (largely accumulated by cells but mostly restricted to lysosomes), and sparfloxacin (accumulated to a fair extent but detected only in cytosol). We used a continuous line of myelomonocytic cells (THP-1 macrophages), which display specific surface receptors for IFN-gamma, and examined the activity of these antibiotics against L. monocytogenes Hly+ (virulent variant) and L. monocytogenes Hly- (a nonvirulent variant defective in hemolysin production). Untreated THP-1 and phorbol myristate acetate differentiated THP-1 were permissive for infection and multiplication of intracellular L. monocytogenes Hly+ (virulent variant). All three antibiotics tested were bactericidal against this Listeria strain when added to an extracellular concentration of 10x their MIC. After preexposure of THP-1 to IFN gamma, L. monocytogenes Hly+ was still phagocytosed but no longer grew intracellularly. The activity of ampicillin became almost undetectable (antagonistic effect), and that of azithromycin was unchanged (additive effect with that of IFN-gamma), whereas that of sparfloxacin was markedly enhanced (synergy). A similar behavior (lack of bacterial growth, associated with a loss of activity of ampicillin, an enhanced activity of sparfloxacin, and unchanged activity of azithromycin) was observed in cells infected with L. monocytogenes Hly-. This modulation of antibiotic activity, which we ascribe to the change of subcellular localization of L. monocytogenes caused by IFN-gamma or by the lack of virulence factor, could result from a change in bacterial responsiveness to antibiotics, a modification of the drug activity, or differences in drug bioavailabilities between cytosol and phagosomes. PMID- 8723472 TI - Zidovudine, trimethoprim, and dapsone pharmacokinetic interactions in patients with human immunodeficiency virus infection. AB - Zidovudine is widely prescribed for the treatment of human immunodeficiency virus (HIV) infection. Trimethoprim and dapsone are commonly used in the management of Pneumocystis carinii pneumonia in HIV-infected patients. To examine the pharmacokinetic interactions among these drugs, eight HIV-infected patients (26 to 43 years old) with a mean CD4 count of 524.4 +/- 405.7 cells per mm3 received zidovudine (200 mg), trimethoprim (200 mg), and dapsone (100 mg) as single agents and in two- and three-drug combinations. Blood and urine samples were collected at a specified time and analyzed for zidovudine, zidovudine-glucuronide, trimethoprim, dapsone, and monoacetyl-dapsone concentrations under single-dose and steady-state conditions. Zidovudine did not influence the pharmacokinetic disposition of dapsone or trimethoprim. Dapsone had no effect on the pharmacokinetic disposition of zidovudine. Trimethoprim significantly decreased the renal clearance of zidovudine by 58% (5.0 +/- 1.8 versus 2.1 +/- 0.5 ml/min/kg of body weight [P < 0.05]). There was a concurrent 54% decrease in the mean urinary recovery of zidovudine (11.7 +/- 3.5 versus 5.4 +/- 3.0 [P < 0.05]), and the metabolic ratio was decreased by 78% (0.32 +/- 0.4 versus 0.07 +/- 0.05 [P < 0.05]). The mean area under the concentration-time curve from 0 to 6 h of the zidovudine-glucuronide/ zidovudine ratio was unchanged. We conclude that zidovudine, trimethoprim, and dapsone can be coadministered to patients with AIDS without significant pharmacokinetic interaction. However, in AIDS patients with liver impairment and impaired glucuronidation, doses of zidovudine may need to be decreased. PMID- 8723473 TI - Effect of polyaspartic acid on pharmacokinetics of gentamicin after single intravenous dose in the dog. AB - The effects of poly-L-aspartic acid on the pharmacokinetics of gentamicin were examined by using a randomized crossover trial design with the dog. When analyzed according to a three-compartment open model, poly-L-aspartic acid reduced some first-order rate equation constants (A3, lambda 1, and lambda 3), the deep peripheral compartment exit microconstant (k31), the elimination rate constant (k(el)), and the area under the concentration-time curve from 0 to 480 h (AUC0 480) (0.21-, 0.60-, 0.26-, 0.27-, 0.72-, and 0.76-fold, respectively; P < 0.05) but increased the volume of distribution at steady state (Vss), the volume of distribution calculated by the area method (V(area)), the apparent volume of the peripheral compartment (Vp), and all mean time parameters. These results suggested that poly-L-aspartic acid increased the distribution of gentamicin to or binding within the deep peripheral compartment and that poly-L-aspartic acid may have delayed gentamicin transit through the peripheral tissues. In contrast, poly-L-aspartic acid did not alter pharmacokinetic parameters relevant to the central or shallow peripheral compartments to a clinically significant extent. Although gentamicin's pharmacokinetic parameters of relevance to therapeutic drug monitoring were not directly altered, this study has provided pharmacokinetic evidence that poly-L-aspartic acid alters the peripheral distribution of gentamicin. This pharmacokinetic interaction occurred after a single intravenous dose of each drug. Therefore, this interaction should be investigated further, before polyaspartic acid can be considered for use as a clinical nephroprotectant. PMID- 8723475 TI - In vitro activity of the tricyclic beta-lactam GV104326. AB - GV104326 is a novel tricyclic beta-lactam (a trinem or, formerly, tribactam). The in vitro activity of GV104326 was compared with those of cefuroxime, cefixime, amoxicillin, amoxicillin-clavulanic acid, cefpirome, and ciprofloxacin. GV104326 had in vitro activity generally similar to that of cefixime against members of the family Enterobacteriaceae (MIC at which 90% of the isolates are inhibited [MIC90], < or = 2 micrograms/ml), with cefuroxime and amoxicillin-clavulanic acid being 8- to 32-fold less active and with cefpirome being 4- to 8-fold more active against members of this family. The trinem had no activity against Pseudomonas aeruginosa or Stenotrophomonas maltophilia (MIC90, > 128 micrograms/ml) but was the most active agent against Acinetobacter calcoaceticus. GV104326 was particularly active against gram-positive cocci. Ninety percent of methicillin susceptible Staphylococcus aureus strains were susceptible to 0.03 microgram of GV104326 per ml, making it the most active agent studied. Enterococci and Lancefield group A and B streptococci were generally equally or somewhat more susceptible to GV104326 than they were to amoxicillin. Streptococcus pneumoniae strains were highly susceptible to GV104326, and those strains which showed decreased susceptibility to penicillin were generally twofold more susceptible to the trinem than to amoxicillin. Haemophilus influenzae and Moraxella catarrhalis were highly susceptible to GV104326 (MIC90s, 0.12 and 0.03 microgram/ml, respectively). The anaerobes Clostridium perfringens, Bacteroides fragilis, and Peptostreptococcus spp. were more susceptible to the trinems (formerly tribactams) than to the other agents studied. PMID- 8723474 TI - Population pharmacokinetic study of teicoplanin in severely neutropenic patients. AB - The teicoplanin pharmacokinetics (PK) of 30 febrile and severely neutropenic patients (polymorphonuclear count, < 500/mm3) with hematologic malignancies were compared with those determined for five healthy volunteers (HV). Neutropenic patients were given piperacillin combined with amikacin, and teicoplanin was added to the regimen the day fever developed in patients suspected of having a staphylococcal infection or 48 h later. Teicoplanin was given intravenously at a dosage of 6 mg/kg of body weight at 0, 12, and 24 h and once a day thereafter. Five to eleven blood samples per patient were collected. Teicoplanin concentrations were measured by liquid chromatography. A bicompartmental model was fitted to the data by a nonlinear mixed-effect-model approach. Multiple linear regression analysis was applied in an attempt to correlate PK parameters to nine covariates. The mean trough concentrations of teicoplanin 48 h after the onset of treatment and 24 h after the last injection (last trough) +/- standard deviations were 8.8 +/- 4.1 and 17.5 +/- 13.5 mg/liter, respectively. A significant increase was noted in the mean rate of elimination clearance of teicoplanin in neutropenic patients compared with that of HV (0.86 versus 0.73 liter/h, P = 0.002), as was the case with rates of distribution clearance (5.89 versus 4.94 liter/h, P = 0.002); the mean half-life of distribution was significantly shorter in patients than in HV (0.43 versus 0.61 h, P = 0.002). In contrast, the volumes of the central compartment (ca. 5.8 liters for both groups), the volumes of distribution at steady state (HV, 37.6 liters; patients, 55.9 liters), and the elimination half-lives (HV, 39.6 h; patients, 52.7 h) were not significantly different between HV and neutropenic patients. Interindividual variabilities of rates of clearance (coefficient of variation [CV], 43%) and elimination half-lives (CV, 56%) were mainly explained by the variabilities among rates of creatinine clearance. Interindividual variabilities of the volumes of the central compartment (CV, 33%) and the volumes of distribution at steady state (CV = 51%) were correlated to interindividual variabilities among numbers of leukocytes and the ages of patients, respectively. On the basis of the population PK model of teicoplanin, simulations were made to optimize the dosing schedule. A supplemental 6 mg/kg dose of teicoplanin at 36 h resulted in a trough concentration at 48 h of 16.0 +/- 4.5 mg/liter, with only 7% of patients having a trough concentration of less than 10 mg/liter, compared with 46% of patients on the usual schedule. PMID- 8723476 TI - Sequence and characterization of a novel chromosomal aminoglycoside phosphotransferase gene, aph (3')-IIb, in Pseudomonas aeruginosa. AB - A novel, probably chromosomally encoded, aminoglycoside phosphotransferase gene was cloned on a 2,996-bp PstI fragment from Pseudomonas aeruginosa and designated aph (3')-IIb. It coded for a protein of 268 amino acids that showed 51.7% amino acid identity with APH (3')-II [APH(3') is aminoglycoside-3' phosphotransferase] from Tn5. Two other open reading frames on the cloned fragment showed homology to a signal-transducing system in P. aeruginosa. PMID- 8723477 TI - Directly repeated insertion of 9-nucleotide sequence detected in penicillin binding protein 2B gene of penicillin-resistant Streptococcus pneumoniae. AB - We investigated the molecular mechanism of 50 penicillin-resistant Streptococcus pneumoniae strains (penicillin: MIC, > or = 0.125 microgram/ml) having neither class A nor class B mutations in the penicillin-binding protein 2B gene (pbp2b). An analysis of the nucleotide sequences of the pbp2b genes from seven strains revealed an unique direct repeat of 9 nucleotides (TGGTATACT) between active-site serine (residue 385) and Ser-X-Asn (residues 442 to 444) motifs. The same insertion was detected in 13 strains. PMID- 8723478 TI - Increasing antibiotic resistance in clinical isolates of Aeromonas strains in Taiwan. AB - A total of 234 clinical isolates of Aeromonas, primarily A. hydrophila, were collected for the present study. Most were isolates from blood. By the agar dilution method, more than 90% of the Aeromonas strains were found to be susceptible to moxalactam, ceftazidime, cefepime, aztreonam, imipenem, amikacin, and fluoroquinolones, but they were more resistant to tetracycline, trimethoprim sulfamethoxazole, some extended-spectrum cephalosporins, and aminoglycosides than strains from the United States and Australia. PMID- 8723479 TI - Plasmid-borne high-level resistance to gentamicin in Enterococcus hirae, Enterococcus avium, and Enterococcus raffinosus. AB - Enterococcus hirae, E. avium, and E. raffinosus isolated in Romania, Tunisia, and Portugal harbored plasmids pICC8, pIP1700, and pIP1701, respectively, encoding resistance to high levels of gentamicin (Gmr). The Gmr marker was carried on pIP1700 by a Tn4001-like element and on pICC8 and pIP1701 by Tn4001-truncated structures. pICC8 carried, in addition to Gmr, chloramphenicol, erythromycin, and tetracycline-minocycline (TetM) resistance determinants. The gene tetM of pICC8 was carried on a Tn916-like element. PMID- 8723480 TI - Multiple antibiotic resistance (mar) locus protects Escherichia coli from rapid cell killing by fluoroquinolones. AB - The multiple antibiotic resistance (mar) locus in Escherichia coli consists of two divergently expressed operons (marC and marRAB), both of which contribute to the Mar phenotype. Overexpression of the marRAB operon protected E. coli against rapid cell killing by fluoroquinolones. Inactivation of the operon in mar mutants restored a wild-type bactericidal susceptibility. Both operons of the locus were required for protection from the quinolone-mediated bactericidal activity in mar locus deletion mutants. The effect was lost at high concentrations of fluoroquinolones, unlike the case for the previously described genes hipA and hipQ. The inducible mar locus appears to specify a novel antibactericidal mechanism which may play a role in the emergence of fluoroquinolone-resistant clinical E. coli isolates. PMID- 8723481 TI - Comparative kinetic analyses of interaction of inhibitors with Rauscher murine leukemia virus and human immunodeficiency virus reverse transcriptases. AB - The inhibitory effects of several nucleoside triphosphate analogs on Rauscher murine leukemia virus (RMuLV) and human immunodeficiency virus (HIV) type 1 reverse transcriptases (RTs) were studied. With RNA as the template, the apparent K(m) and apparent K(i) values of HIV RT toward its substrates and inhibitors are 12 to 500 times lower than the corresponding values for RMuLV RT. However, the k(i)/k(m) ratios (inhibition efficiencies) for HIV and RMuLV RTs'are similar for AZTTP (zidovudine triphosphate), d4TTP [3'-deoxythymidine-2'-ene-(3'-deoxy-2',3' didehydrothymidine) triphosphate], PMEADP [9-(2-phosphonylmethoxyethyl)adenine diphosphate], FIAUTP [1-(2-fluoro-2-deoxy-beta-D-arabinofuranosyl)-5-iodouracil triphosphate], and HPMPCDP [(S)-1-(3-hydroxy-2-phosphylmethoxypropyl) cytosine diphosphate]. With DNA as the template, the K(m) values are similar for HIV and RMuLV RTs. However, the K(i)/K(m) values of HIV and RMuLV RTs are significantly different for ddCTP, ddATP, and 3TCTP (2',3'-dideoxy-3'-thiacytidine). The RTs of RMuLV and HIV are sufficiently different from one another that the kinetic inhibition constants for a particular antiviral compounds should be determined to indicate whether anti-RMuLV activity is likely to be predictive for the anti-HIV activity of the compound. This information, in conjunction with species-specific drug metabolism differences and tissue culture antiviral activity, is important in determining the suitability of a particular animal model. PMID- 8723482 TI - N-acetylcysteine inhibits germination of conidia and growth of Aspergillus spp. and Fusarium spp. AB - N-Acetylcysteine inhibited hyphal growth and germination of conidia of Aspergillus spp. and Fusarium spp. N-Acetylcysteine inhibited conidial germination as well as or better than L-cysteine. Cysteine-related compounds may provide a potential therapeutic strategy against agriculturally and medically important fungal pathogens. PMID- 8723484 TI - Ceftazidime kinetics of diffusion in inoculated agar plates. AB - The ceftazidime concentration in agar plates inoculated with Pseudomonas aeruginosa was determined by high-performance liquid chromatography at fixed points (3, 6, 9, 12, and 15 mm) from the disk center and at fixed times (2, 4, 6, 16, and 24 h) to study the antibiotic kinetics of diffusion. A statistical difference between the concentrations determined in the presence of microorganisms and in uninoculated plates after 16 and 24 h was evidenced and was probably ascribable to the drug hydrolysis carried out by the induced beta lactamase. PMID- 8723483 TI - In vitro activities of semisynthetic pneumocandin L-733,560 against fluconazole resistant and -susceptible Candida albicans isolates. AB - Lipopeptide L-733,560 is a water-soluble derivative of pneumocandin B0 that exhibits enhanced anti-Candida activity. We investigated the in vitro activity of L-733,560 compared with those of amphotericin B, flucytosine, and itraconazole, against fluconazole-resistant (n = 44) and fluconazole-susceptible (n = 46) Candida albicans isolates. Tests were performed with a photometer-read broth microdilution method with RPMI-2% glucose and National Committee for Clinical Laboratory Standards reference strains. Except for those of itraconazole, MICs were not significantly different between the two groups of isolates, as expected for agents with different mechanisms of action. L-733,560 was the most active agent against C.albicans, with MICs for 50 and 90% of the strains tested of 0.01 and 0.06 microgram/ml, respectively. PMID- 8723485 TI - Penciclovir is a selective inhibitor of hepatitis B virus replication in cultured human hepatoblastoma cells. AB - Penciclovir [9-(4-hydroxy-3-hydroxymethylbut-1-yI)guanine], an effective antiherpesvirus agent, was found to be a potent and selective antiviral agent against intracellular hepatitis B virus (HBV) replication (drug concentration at which a 10-fold decrease in HBV DNA from the average level in an untreated culture was observed [EC90], 1.6 microM) and extracellular virion release (EC90, 0.7 microM) by cultured human hepatoblastoma (2.2.15) cells. Acyclovir and three other related 9-alkoxypurines with activity against either herpesviruses or human immunodeficiency virus were uniformly inactive against HBV. The activity of penciclovir is discussed in relation to recent findings related to its mode of action against HBV. PMID- 8723486 TI - Synergistic inhibition of human immunodeficiency virus isolates (including 3' azido-3'-deoxythymidine-resistant isolates) by foscarnet in combination with 2',3'-dideoxyinosine or 2',3'-dideoxycytidine. AB - The combinations of foscarnet plus 2',3'-dideoxyinosine and foscarnet plus 2',3' dideoxycytidine synergistically inhibit the replication of human immunodeficiency virus type 1 isolates, including two 3'-azido-3'-deoxythymidine-resistant isolates. The combination of 2',3'-dideoxyinosine plus 2',3'-dideoxycytidine showed additive inhibition of the majority of the human immunodeficiency virus isolates tested. All three combinations showed pronounced antagonistic cytotoxicity and thus were less toxic to the growth of peripheral blood mononuclear cells than the separate drugs. PMID- 8723487 TI - Differences in the resistant variants of Enterobacter cloacae selected by extended-spectrum cephalosporins. AB - The rates of development of resistance to ceftriaxone, ceftazidime, cefepime, and cefpirome in 10 strains of Enterobacter cloacae were determined by daily transfer for 7 days to fresh medium containing twofold serial dilutions of the antibiotics. Development of resistance to ceftriaxone was the most rapid; this was followed by ceftazidime, cefpirome, and cefepime. Resistant variants selected by ceftriaxone and ceftazidime were cross-resistant and produced very high levels of beta-lactamase. On the other hand, resistant variants selected by cefepime and cefpirome often had moderately high levels of beta-lactamase and diminished levels of the 39- to 40-kDa porin protein. PMID- 8723488 TI - In vitro and in vivo immunomodulatory effects of anti-Pneumocystis carinii drugs. AB - The anti-Pneumocystis carinii drug effects on mitogen-, antigen-, and interleukin 2-induced proliferative responses and on natural killer (NK) cell-mediated activity were analyzed in vivo (rats) and in vitro (normal human peripheral blood mononuclear cells). Splenocytes derived from in vivo piritrexim- and clindamycin treated rats showed a significant inhibition of mitogen-induced proliferative responses. In vitro exposure to clindamycin, piritrexim, and pyrimethamine caused an inhibition of human T lymphocyte proliferation in response to mitogen, antigen, and interleukin-2 stimulation. Rat NK cell-mediated cytotoxic activity was not affected by the drugs, and human NK cell activity was reduced only at the highest concentration (10 micrograms/ml) of the drugs. The potential immunotoxicity of the long-term administration of these agents in humans needs further investigation. PMID- 8723489 TI - Trophozoite elimination in a rat model of Pneumocystis carinii pneumonia by clinically achievable plasma deferoxamine concentrations. AB - In a rat model of Pneumocystis carinii pneumonia, a 3-week infusion of deferoxamine producing concentrations in plasma of > or = 1.5 micrograms m-1 eliminated the trophozoite life cycle stage. Since this concentration is well below that routinely achieved in patients treated for iron overload, deferoxamine has promise as a therapy for AIDS-associated P.carinii pneumonia. PMID- 8723490 TI - 50S ribosomal subunit synthesis and translation are equivalent targets for erythromycin inhibition in Staphylococcus aureus. AB - Macrolide antibiotics like erythromycin can prevent the formation of the 50S ribosomal subunit in growing bacterial cells, in addition to their inhibitory effect on translation. The significance of this novel finding has been further investigated. The 50% inhibitory doses of erythromycin for the inhibition of translation and 50S subunit assembly in Staphylococcus aureus cells were measured and were found to be identical. Together they account quantitatively for the observed effects of erythromycin on cell growth rates. There is also a direct relationship between the loss of rRNA from the 50S subunit and its accumulation as oligoribonucleotides in cells. The importance of this second site for erythromycin inhibition of bacterial cell growth is discussed. PMID- 8723491 TI - Anti-influenza virus activities of 4-substituted 2,4-dioxobutanoic acid inhibitors. AB - We previously identified a series of compounds which specifically inhibited the transcription of influenza A and B viruses (J. Tomassini, H. Selnick, M.E. Davies, M.E. Armstrong, J. Baldwin, M. Bourgeois, J. Hastings, D. Hazuda, J. Lewis, W. McClements, G. Ponticello, E. Radzilowski, G. Smith, A. Tebben, and A. Wolfe, Antimicrob. Agents Chemother. 38:2827-2837, 1994). The compounds, 4 substituted 2,4-dioxobutanoic acids, selectively targeted the cap-dependent endonuclease activity of the transcriptase complex. Additionally, several of these compounds effectively inhibited the replication of influenza virus but not other viruses in cell culture assays. Here, we report on the anti-influenza virus activities of other potent derivatives of the series evaluated in both in vitro and in vivo infectivity assays. These compounds inhibited the replication of influenza virus in yield reduction assays, with 50% inhibitory concentrations ranging from 0.18 to 0.71 microM. These 50% inhibitory concentrations were similar to those observed for inhibition of in vitro transcription (0.32 to 0.54 microM). One selected compound also elicited a dose-dependent inhibition of influenza virus replication in mice following an upper respiratory tract challenge. These studies demonstrate the antiviral efficacy of this inhibitor class and thereby establish the utility of influenza virus endonuclease as a chemotherapeutic target. PMID- 8723492 TI - Treatment of Staphylococcus aureus catheter-related infection and infective endocarditis with granulocyte colony-stimulating factor in the experimental rabbit model. AB - The role of granulocyte colony-stimulating factor with and without antibiotics in the treatment of catheter-related infection and infective endocarditis caused by methicillin-susceptible Staphylococcus aureus was assessed in the experimental rabbit model. Granulocyte colony-stimulating factor stimulated leukocytosis in infected animals but did not increase the clearance of methicillin-susceptible S. aureus from peripheral blood, subcutaneous port catheters, intravascular cardiac catheters, or aortic valve vegetations. PMID- 8723493 TI - Concentrations of ceftriaxone (1,000 milligrams intravenously) in abdominal tissues during open prostatectomy. AB - Ceftriaxone concentrations in abdominal tissues were evaluated at different stages of open prostatectomy. Ceftriaxone was administered as antibiotic prophylaxis, and 15 consecutive patients were given a single dose of ceftriaxone (1,000 mg intravenously in 1 min) 30 min before surgery. Ceftriaxone concentrations in tissue were determined at three stages of the surgical procedure; upon the opening of the abdominal cavity, during the prostatectomy, and upon the closure of the abdominal cavity. Samples of the following tissues or sample were assayed: epiploic and abdominal-wall fat; Retzius' space, bladder, and prostate tissue; and urine. During the different stages of the surgical procedure, for all patients, and in the different tested tissues, ceftriaxone concentrations greater than or equal to the cutoff point (4 micrograms/g of tissue) were measured. The highest concentrations were obtained in the bladder (43 +/- 18 micrograms/g) and in the prostate (35 +/- 18 micrograms/g). In fatty tissues, concentrations were between 13 +/- 5 and 22 +/- 8 micrograms/g. All patients (15 of 15) had ceftriaxone levels in tissue greater than the MICs for the potential pathogens (Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, and Proteus mirabilis). In conclusion, during open prostatectomy and after the use of a single dose of ceftriaxone (1,000 mg), high antibiotic levels were obtained throughout the surgical procedure in the tissues potentially involved in postoperative infection. PMID- 8723494 TI - In vitro and in vivo activities of SCH 56592 against Blastomyces dermatitidis. AB - The new triazole derivative SCH 56592 has been tested in a National Committee for Clinical Laboratory Standards-adapted in vitro susceptibility test, and its activity against 12 isolates of Blastomyces dermatitidis yeast-like forms has been compared with those of amphotericin B, itraconazole, and fluconazole. SCH 56592 was the most active of the four compounds, with an MIC at which 90% of the isolates are inhibited of 0.06 microgram/ml and a minimal fungicidal concentration at which 90% of the isolates are inhibited of 4 micrograms/ml. The results of the treatment of mice infected with B. dermatitidis with three different doses of SCH 56592 (25, 5, or 1 mg/kg of body weight), amphotericin B (1 mg/kg), or itraconazole (150 mg/kg) confirmed the potent activity of SCH 56592. Survival was prolonged at each dose of SCH 56592, and sterilization of the lungs occurred in the high-dose group but not in the groups treated with itraconazole or fluconazole. SCH 56592 is a promising new azole antifungal drug that should be studied in humans with blastomycosis. PMID- 8723495 TI - Variation in fluconazole efficacy for Candida albicans strains sequentially isolated from oral cavities of patients with AIDS in an experimental murine candidiasis model. AB - Four strains of Candida albicans, isolated from two patients with AIDS who had undergone prolonged fluconazole therapy for oral candidiasis, were studied in a model of disseminated murine candidiasis. Pre- and posttreatment isolates from each patient were genetically related, and the fluconazole MICs for the strains had increased significantly, from 0.25 to 32 micrograms/ml for the strains isolated from patient 1 and from 1.0 to 16 micrograms/ml for the strains isolated from patient 2. Mice were infected intravenously and were treated orally with fluconazole. For survival studies, mice were treated from day 1 to day 10 postinfection and were observed through day 30. The fluconazole dosages were as follows: 0.25, 0.5, 1.0, and 5.0 mg/kg of body weight twice a day. For tissue burden studies, two groups of mice (each group received fluconazole at 0.25 or 5.0 mg/kg) were treated from day 1 to day 7 and were sacrificed 1 day later for quantitative tissue cultures of the spleen and both kidneys. For pretreatment isolates from both patients, all fluconazole dosing regimens were effective at prolonging survival compared with the survival of the control groups. For posttreatment isolates, only fluconazole at 5.0 mg/kg was effective at prolonging survival. Both fluconazole dosing regimens used in the tissue burden studies significantly reduced the counts of the pretreatment isolate from patient 1 in the spleen and kidney, while fluconazole at 5.0 mg/kg was effective at reducing the counts of the posttreatment isolate. For both isolates from patient 2, only fluconazole at 5.0 mg/kg was effective at reducing the counts in the spleen and kidney. The study indicates that C. albicans mutation to resistance to fluconazole may play a critical role in fluconazole-refractory oral candidiasis in AIDS patients. PMID- 8723496 TI - Kill kinetics and regrowth patterns of Pseudomonas aeruginosa exposed to concentration-time profiles of tobramycin simulating in vivo infusion and bolus dosing. AB - Pseudomonas aeruginosa ATCC 27853 was exposed to tobramycin concentration-time profiles modelling in vivo bolus and infusion dosing. Dependence of bactericidal and bacteriostatic activity on the initial profile of peak concentration (bolus effect > infusion) and area under the antibiotic concentration-time curve was observed at peak concentration/MIC ratios of 10 or below. PMID- 8723497 TI - Cefuroxime efficacy in treatment of bacteremic pneumonia due to penicillin resistant and cefuroxime-resistant Streptococcus pneumoniae. PMID- 8723498 TI - Is bactericidal activity of amoxicillin against Helicobacter pylori concentration dependent? PMID- 8723499 TI - Potentiation of the stavudine anti-human immunodeficiency virus activity by 5 fluorouracil. PMID- 8723500 TI - Radiochemistry and radiopharmacy. PMID- 8723501 TI - Clinical radiopharmacy: principles and practices. AB - On the average, radiopharmacists spend about 17.2% of their time in clinical activities if their practice setting is in an institution, and about 8.5% of their time if their practice setting is in a centralized nuclear pharmacy. A recent survey of radiopharmacists was conducted to determine: (1) the percentage of time they spend engaged in selected activities, and (2) the specific clinical activities in which they are involved. A few radiopharmacists spend as much as 50% of their time in clinical activities, but most spend only 5% to 20% of their time. Some of the clinical activities involve direct interactions with patients, such as explaining the reasons for administering the radioactive material or actually administering the dose. Other clinical activities are indirect, such as reviewing charts before or after studies and making recommendations to other health care professionals. About half of the pharmacists surveyed see a need for increasing their clinical activities. The need to maximize the time involved in providing pharmaceutical care is discussed and several patient-care activities/responsibilities are proposed. PMID- 8723502 TI - The role of commercial nuclear pharmacy in the future practice of nuclear medicine. AB - It has been estimated that today 70% to 80% of all radiopharmaceutical doses are dispensed through commercial nuclear pharmacy channels. These services are provided by the approximately 250 facilities in the United States, with some multisite corporations dispensing in excess of 20,000 unit-dose prescriptions per day. As pressures mount within health care institutions to reduce manpower, increase cost-effectiveness, increase participation in managed care contracts, and to seek outside vendors for many services that were previously provided in house, the future role of the commercial nuclear pharmacy in the practice of nuclear medicine will only continue to increase. The essence of nuclear pharmacy practice is the dispensing of a full range of high quality radiopharmaceuticals in patient-specific unit doses. These doses must be delivered in a timely and cost effective manner, without compromising quality or patient safety. Commercial nuclear pharmacies have expanded to provide such varied functions as radiation safety and waste management, as well as consultative and marketing activities directed towards clinicians within a nuclear medicine practitioners own facility. In-service continuing education programs directed towards physicians and technologists are frequently offered by many commercial nuclear pharmacies. Changes in health care economics, merging and down-sizing in the hospital industry, and the overall impact of managed care on the viability of hospitals in general has resulted in slow growth, or even a small decline in the number of institutionally based nuclear pharmacists. As a result, nuclear medicine practitioners will be looking to the commercial nuclear pharmacies to meet a larger portion of their radiopharmaceutical needs, as well as to value added services, such as education and research and development. Specialized practice settings, such as nuclear cardiology and free-standing nuclear medicine clinics, are especially well suited to the services provided by commercial nuclear pharmacies. Involvement in the distribution of positron-emission tomography radiopharmaceuticals will continue to increase regardless of the results of current regulatory debates on this issue. In the future, nuclear medicine practitioners will look to the commercial nuclear pharmacies for an increasing portion of their radiopharmaceutical needs and the industry should be ready and able to meet these demands in a safe, timely, and cost efficient manner. PMID- 8723503 TI - Radiopharmaceuticals as orphan drugs. AB - Rare diseases and conditions are defined as diseases or conditions that affect less than 200,000 individuals in the United States. Because of the limited population suffering from any given rare disorder, there is little economic motivation for the development of drug products for its diagnosis or treatment. Also, frequently these rare diseases are misdiagnosed or undiagnosed due to a lack of familiarity with their symptoms and course. Thus, these rare disorders are commonly referred to as orphan diseases or conditions. To address this problem, legislation has been enacted that provides for incentives for the development of drug products for rare diseases or conditions (i.e., orphan drug products). Nuclear medicine scientists can take advantage of these incentives in the development of orphan radiopharmaceuticals for the treatment or diagnosis of rare diseases or conditions. Nuclear medicine physicians should know where information on orphan diseases and orphan radiopharmaceuticals can be obtained to maximize the care of their patients. PMID- 8723504 TI - Radiopharmaceutical regulation and Food and Drug Administration policy. AB - The regulatory policy of the Food and Drug Administration (FDA) on radiopharmaceuticals flows from a rigid, traditional, drug-like interpretation of the FDC Act on the licensing of radiopharmaceuticals. This contributes to significant delays in the drug-approval process for radiopharmaceuticals, which are very costly to the nuclear medicine community and the American public. It seems that radiopharmaceuticals would be better characterized as molecular devices. Good generic rule-making principles include: use of a risk/benefit/cost analysis; intent based on sound science; performance standards prepared by outside experts; a definite need shown by the regulatory agency; to live with the consequences of any erroneous cost estimates; and design individual credential requirements so that additional training results in enhanced professional responsibility. When these common elements are applied to current FDA policy, it seems that the agency is out of sync with the stated goals for revitalizing federal regulatory policies as deemed necessary by the Clinton administration. Recent FDA rulings on positron-emission tomography, Patient Package inserts, and on medical device service accentuate the degree of such asynchronization. Radiopharmaceutical review and licensing flexibility could be dramatically improved by excluding radiopharmaceuticals from the drug category and reviewing them as separate entities. This new category would take into account their excellent record of safety and their lack of pharmacological action. Additionally, their evaluation of efficacy should be based on their ability to provide useful scintiphotos, data, or responses of the physiological system it portends to image, quantitate, or describe. To accomplish the goal of transforming the FDA's rigid, prescriptive policy into a streamlined flexible performance-based policy, the Council on Radionuclides and Radiopharmaceuticals proposal has been presented. In addition, it is suggested that the United States Pharmacopeia write radiopharmaceutical review standards, that an independent scientific body review the data submitted, and that the FDA either accept or reject the recommendation. PMID- 8723505 TI - Radiation safety in the central radiopharmacy. AB - The currently increasing number of new radiopharmaceuticals and the increase in individual patient-dose activity, particularly for single photon emission computer tomography imaging, coupled with the switch toward unit doses probably contribute to an increase in the radiation burden to radiopharmacy personnel. To meet this increased need of the nuclear medicine community, a medium-sized radiopharmacy may order seven to eight 3-Ci molybdenum-99/technetium-99M generators per week and may perform 10 to 15 elutions per day. The maximum radiation exposure to radiopharmacy personnel occurs during the generator elutions, compounding (kit preparation), and unit-dose dispensing. Additional sources of radiation exposure can occur during receipt of radioactive packages, transport of prepared doses, and the process of storage and disposal of radioactive waste. Recent reports have shown extremity radiation doses to the fingers and hands at 14 to 21 rem/year. Radiation exposure can be monitored by film badges, thermoluminescent detectors, or one of the analog or digital self reading dosimeters. Each monitoring device has advantages and the choice depends upon the personnel involved (eg, visitors) and the particular needs of the radiopharmacy. Extremity dose reductions during radiopharmacy procedures can be achieved by a number of techniques, some as simple as the use of 8- to 9-inch forceps during transfers of radioactive vials and syringes and the use of a dose drawing syringe shield during unit-dose dispensing procedures. In preparation of unsealed doses or capsules of radioactive iodine, there is a potential for a high internal dose because of inhalation of the volitile solution. Frequent monitoring in the form of bioassays and the use of strict threshold levels will ensure early detection of exposure to radioactive iodine and quick corrective action. In pharmacies and nuclear medicine departments handling fluorine-18 for use with Anger gamma cameras, the high levels of both beta and gamma radiation present an additional set of radiation safety problems. PMID- 8723506 TI - Is there life after technetium: what is the potential for developing new broad based radionuclides? AB - The use of radionuclides for medical and for a multitude of other basic research applications has continued to grow at a very rapid pace. Procedures, based on their use as radiotracers for nuclear medicine imaging and for radiotherapy of cancer and other pathology, have become firmly established as important clinical modalities. It is estimated that on an annual basis in the United States alone, radionuclides are used medically in over 13 million imaging procedures, in over 100 million laboratory tests, and in an ever increasing number (> 100,000) for therapeutic administrations. One out of every four hospital patients undergoes a procedure that involves the use of radionuclides. Diagnostic imaging methods using planar/single-photon emission computed tomography and positron-emission tomography (PET) imaging, as well as the measurement of in vivo organ function, physiology, or biochemistry, have become indispensable tools in patient workup and management. More than 80% of all imaging studies (mostly anatomic) currently use technetium-99m (99mTc), because it has turned out to be the ideal isotope from various considerations. However, over the past few years, nuclear medicine has experienced a slow but steady evolution towards functional studies, quantitative PET imaging, and novel therapeutic approaches. New radionuclides are required for these applications, and their development has attracted considerable interest. This article reviews the current status and future prospects for the development of many new potential isotopes. Practical issues, such as the feasibility of large-scale production and wide-spread availability in a continuous reliable fashion, are addressed. To date, the data are not sufficient to answer the question as to whether any of these radionuclides (or their applications, for that matter) will eventually assume as broad-based a role as that of 99mTc. Nonetheless, there are a number of promising radionuclides that could assume an important place in the future practice of nuclear medicine. PMID- 8723507 TI - Widespread unilateral increased rib uptake on bone scan. PMID- 8723508 TI - Noncardiac uptake of technetium-99m sestamibi: an updated gamut. PMID- 8723509 TI - Liver visualization on perfusion scan after umbilical vein catheter administration. PMID- 8723510 TI - Ultrasound-guided fine-needle aspiration biopsy of lung cancers. AB - One hundred eighty-eight patients with 191 lung cancers were collected retrospectively to evaluate the diagnostic results and to determine the accuracy of cytologic diagnoses obtained from ultrasound-guided fine-needle aspiration biopsy (US-guided FNAB), and to discuss the necessity of large-bore tissue core needle biopsy. All 188 patients underwent US-guided FNAB, and 20 patients with 21 lung tumors also underwent US-guided tissue-core needle biopsy. Using US-guided FNAB alone, the positive cytologic results and correct cytologic diagnoses were 91% (174 of 191) and 71% (37 of 52). If both US-guided FNAB and selected US guided tissue core needle biopsy (n = 21) were evaluated, the positive cytologic or histologic results and correct cytologic or histologic diagnoses were 94% (180 of 191) and 80% (45 of 57), respectively. Analyzing the disagreement between the cytologic results and histologic diagnoses (n = 15), we found that the disagreement usually occurred in the specimens with poorly differentiated carcinomas (nonspecific cell type) (53% [8 of 15]); of these, two patients (13% [2 of 15], small cell carcinoma) would have a change in treatment. The complications of US-guided FNAB were pneumothorax (n = 3), hemoptysis (n = 1), hemothorax (n = 1), and suspected tract metastasis (n = 1). We conclude that US guided FNAB has a high diagnostic yield in lung cancers, and US-guided tissue core needle biopsy is only necessary in patients whose cytologic results are negative or who have poorly differentiated carcinomas. PMID- 8723511 TI - Sonography of hepatobiliary ascariasis. AB - We present the real-time sonographic features of 19 surgically proven cases of hepatobiliary ascariasis, in which ultrasound examination was the primary modality used to achieve the diagnosis. All the patients presented with right upper quadrant abdominal symptoms. There were no false positive diagnoses. The "impacted worm" sign, reported here for the first time, presented in two patients as long curved, tubular, non-shadowing structures with echoluscent cores, in the distribution of the intrahepatic biliary ducts. Both of these patients died soon after surgery indicating poor prognosis in such cases. Two other patients are described in whom macerated round worms were seen as intraluminal, localized, soft tissue masses in the common duct mimicking a cholangiocarcinoma. Thus, biliary ascariasis must be added to the differential diagnosis of an intraluminal bile duct mass in patients from endemic areas. PMID- 8723512 TI - Relevance of the determination of ovarian volume in adolescent girls with menstrual disorders. AB - Pelvic ultrasound and hormonal studies were performed in 29 adolescent patients, aged 12 to 20 years, to evaluate menstrual irregularities. Patients were divided in three groups according to ultrasound ovarian volumes: group I (n = 16) both ovaries < 10 cm3; group II (n = 8) one of the ovaries > or = 10 cm3; and group III (n = 5) both ovaries > or = 10 cm3. Serum levels of LH, LH:FSH ratio, testosterone, and androstenedione were significantly higher (p < .05) in group III. Positive predictive value of both ovarian volumes > or = 10 cm3 in terms of polycystic ovary syndrome (PCOS) was 100%, negative predictive value was 81%, sensitivity was 63%, specificity was 100%. These data suggest that, in adolescent patients with menstrual disorders, bilateral ovarian volumes of higher than 10 cm3 are correlated with the diagnosis of PCOS. PMID- 8723513 TI - Factors affecting urethrocystographic parameters in urinary continent women. AB - To evaluate the urethrocystographic changes in different conditions, 154 women were evaluated by using introital sonography. Patients were divided into three groups: group 1 (n = 103) normal, including 10 postmenopausal women; group 2 (n = 46) pregnant, including 16 women in the first trimester, 15 in the second trimester, and 15 in the third trimester; group 3 (n = 15) severe genitourinary prolapse. None of the 154 women had a history of urinary incontinence. The following parameters were measured at rest: urethral thickness, uretheral length, urethral inclination, and posterior urethrovesical angle. On maximum straining, urethral inclination, posterior urethrovesical angle, and rotational angle were measured. In general, age, parity, and menopause did not affect the urethrocystographic parameters in Group 1 patients. Postmenopausal women had a significant decrease in the urethral thickness compared with the premenopausal women (p = 0.026). Patients in Groups 2 and 3 had a significantly lower urethral position than those in group 1. However, hypermobility of the urethra was found only in Group 3. Different menstrual ages did not affect the urethral position but could affect the posterior urethrovesical angle at rest in the first trimester. Introital sonography, without the risk of radiation exposure, enables the observation of static and dynamic changes in the lower urinary tract, both repeatedly and reproducibly. PMID- 8723514 TI - Transvaginal sonohysterography: a new aid in the diagnosis of residual trophoblastic tissue. AB - OBJECTIVE: The study was undertaken to evaluate the diagnostic accuracy of transvaginal sonohysterography in postabortion and postpartum patients suspected of having residual trophoblastic tissue. METHODS: Twenty two women with late postabortion and postpartum bleeding were evaluated by transvaginal sonography and transvaginal sonohysterography. The patients were prospectively divided into two groups according to the sonohysterographic findings. Women with an empty uterine cavity or with free-floating masses received conservative treatment. Those with suspected attached residual trophoblastic tissue underwent curettage. RESULTS: Sonohysterography confirmed the findings of two-dimensional sonography of an empty uterus in 8 patients; however, a free floating endometrial mass was found in 4 patients and that of an empty uterine cavity in another 2 of 14 patients suspected of having residual trophoblastic tissue by the two-dimensional ultrasound. All 14 patients who were treated conservatively ceased bleeding. The 4 patients with a free-floating mass reported ejection of a vaginal mass within 6 days of treatment. Eight patients underwent curettage. Residual trophoblastic tissue was found in all of them. CONCLUSION: Transvaginal sonohysterography is a simple and effective method for the evaluation of residual trophoblastic tissue in patients with postpartum and post-abortion bleeding. Its use may eliminate unnecessary curettage procedures. PMID- 8723515 TI - Control of intrauterine device insertion with three-dimensional ultrasound: is it the future? AB - A comparative study was designed to determine whether three-dimensional transvaginal sonography (3D-TVS) offered advantages over two-dimensional transvaginal sonography (2D-TVS) for the identification and location of IUDs in 66 asymptomatic women. Hysteroscopy was performed in cases in which there was a discrepancy between the information obtained by both methods (n = 14). In eight cases (12.2%) the IUD was misidentified with 2D-TVS. In six cases (9.1%) it was not possible to identify the device model with 2D-TVS. In two cases (3.0%) 2D-TVS failed to identify the position of the device. In contrast, all IUDs were identified and located accurately with 3D-TVS. PMID- 8723517 TI - Superficial dorsal penile vein thrombosis (penile Mondor's phlebitis): ultrasound diagnosis. PMID- 8723516 TI - Imaging of angioleiomyoma. PMID- 8723518 TI - Neuronal cell death in the mammalian nervous system: the calmortin hypothesis. AB - 1. This review is concerned with the calcium-dependent mechanisms involved in neuronal cell death. To this end, it provides definitions of the major types of cell death and then describes what is known of their occurrence during development and degeneration of the mammalian nervous system. 2. An analysis is presented of the different sources and compartments of calcium in neurons and of how these are related to the known calcium-dependent enzymes whose excess activation will lead to cell death. 3. The review uses the relatively large amount of pertinent information now available for other cell types, especially thymocytes, to reveal our limited knowledge of how calcium controls neuronal cell death. 4. In the final section, consideration is given to the identification of those factors that may mitigate against the calcium-dependent pathways leading to neuronal degeneration. PMID- 8723519 TI - Development, pharmacology, role of DT-diaphorase and prospects of the indoloquinone EO9. AB - 1. The indoloquinone EO9 (3-hydroxymethyl-5-aziridinyl-1-methyl-2- (1H-indole-4,7 dione)-propenol; E85/053; NSC 382,459) is a synthetic bioreductive alkylating agent that is structurally related to mitomycin C (MMC). 2. EO9 does, however, show a different mechanism of action and a broader antitumour profile than MMC. It is also a more potent cytotoxic agent in vitro than MMC, probably because of its impressive efficient activation by reductive enzymes, particularly DT Diaphorase. This enzyme is elevated in several tumours compared to normal tissues. 3. The preferential cytotoxicity of EO9 under hypoxic conditions makes it an interesting compound to combine with radiation. 4. In preclinical and the Phase I clinical studies, no myelosuppression was observed but reversible proteinuria was dose-limiting. Phase II clinical studies were started in the summer of 1994. PMID- 8723520 TI - Possible mechanism of antihyperglycemic effect of Azadirachta indica leaf extract. Part IV. AB - 1. Effect of epinephrine on the increment index calculated from intravenous glucose tolerance tests and on hepatic glycogen before and after A. indica leaf extract treatment was studied in normal and streptozotocin-induced diabetic rabbits. 2. A. indica leaf extract, in itself, was found to have no action on peripheral utilization of glucose or on hepatic glycogen. 3. The reduction in peripheral utilization of glucose and glycogenolytic effect due to epinephrine action was blocked by A. indica leaf extract, however, almost completely in diabetic rabbits and to a certain extent in normal ones. 4. The results are discussed. PMID- 8723521 TI - Comparison of the antiplatelet agent potential of the whole molecule, F(ab)2 and Fab fragments of humanized anti-GPIIb/IIIa monoclonal antibody in monkeys. AB - 1. The potential antiplatelet agent use of the whole molecule, F(ab)2 and Fab fragments of humanized antiglycoprotein (GP) IIb/IIIa monoclonal antibody, hC4G1, were investigated in rhesus monkeys. 2. Fab completely inhibited platelet aggregation 1 hr after an i.v. bolus administration of 1 mg/kg without a decrease in platelet count or prolongation of bleeding time, and the duration of inhibition was much shorter than that of F(ab)2. 3. These results suggest that the Fab fragment of hC4G1 may be a more useful antiplatelet agent in patients with acute thromboembolic diseases than the whole molecule or F(ab)2 fragments. PMID- 8723522 TI - Abnormal modulation of cholinergic neurotransmission by opioid in hyperresponsive bronchus of rats. AB - 1. The electrical field stimulation (EFS)-induced bronchoconstriction in vitro in rats challenged by DNP-Ascaris antigen was significantly greater than that in normal rats. 2. Morphine inhibited the EFS-induced bronchoconstriction in normal rats. Whereas the inhibition of EFS-induced bronchoconstriction by the opioid was little, if any, in the DNP-Ascaris-challenged rats. 3. These findings suggest that dysfunction of presynaptic inhibitory modulation through the opioid receptor may take place in the airways of DNP-Ascaris-challenged rats. PMID- 8723523 TI - The reduction of antinociceptive effect of morphine administered intraventricularly is correlated with the decrease of serotonin release from the spinal cord in streptozotocin-induced diabetic rats. AB - 1. The antinociceptive effect of morphine (25 micrograms) administered into the 3rd ventricle was significantly attenuated in streptozotocin-induced diabetic rats as measured by the tail-flick assay. 2. The release of serotonin (5-HT; 5 hydroxytryptamine) from the spinal cord caused by intraventricular injection of morphine (25 micrograms) was significantly reduced in streptozotocin-induced diabetic rats. 3. No differences of 5-HT contents of the spinal cord (lumbar cord) between streptozotocin-induced diabetic- and vehicle-treated rats were found. 4. It is concluded that the reduction of antinociception produced by intraventricular injection of morphine in streptozotocin-induced diabetic rats might be, at least partly, due to the decrease of 5-HT release from the spinal cord. PMID- 8723524 TI - Myocardial and vascular effects of efonidipine in vitro as compared with nifedipine, verapamil and diltiazem. AB - 1. Effects of efonidipine on isolated myocardial and aortic preparations were compared with those of nifedipine, verapamil and diltiazem. 2. All drugs produced concentration-dependent negative chronotropic effects on isolated guinea-pig atrial preparations. The potency order was efonidipine > or = nifedipine > diltiazem > or = verapamil, EC30 values being 3.08 x 10(-8)M, 3.48 x 10(-8)M, 1.27 x 10(-7)M and 1.47 x 10(-7)M, respectively. 3. Nifedipine, verapamil and diltiazem produced concentration-dependent negative inotropic effects on isolated guinea-pig left atrial preparations. The potency order was nifedipine > verapamil > diltiazem, EC30 values being 4.94 x 10(-8)M, 1.49 x 10(-7)M and 8.03 x 10(-7)M, respectively. Efonidipine, even at 1 microM produced no inotropic effect: 10 microM efonidipine decreased the contractile force by about 20%. 4. All drugs concentration-dependently attenuated the KCl-induced contraction of isolated rat aortic ring preparation. The potency order was nifedipine > efonidipine > verapamil > diltiazem, EC30 values being 2.98 x 10(-9)M, 1.24 x 10(-8)M, 3.96 x 10(-8)M and 2.13 x 10(-7)M, respectively. 5. Thus, efonidipine was demonstrated to be a potent vasodilator with negative chronotropic but minimal negative inotropic activity, which may be of benefit in the treatment of cardiovascular disorders. PMID- 8723525 TI - Enhancement of K+ currents by stimulation of protein kinase C in the uterine smooth muscle cells of the pregnant rat. AB - 1. Effects of phorbol esters on the K+ currents in isolated rat uterine smooth muscle cells during (18-day) pregnancy were examined using whole-cell voltage clamp modes. All experiments were performed at room temperature. 2. Test pulses were applied between -20 to + 90 mV from a holding potential of -40 mV. Initially, a transient outward current (ITO) was activated, and outward K+ current (IK) was followed. Threshold potential was - 10 to 0 mV, and the activation was voltage-dependent. At - 80 mV, ITO and IK were 17.8 +/- 3.3 and 13.2 +/- 2.6 pA/pF as a current density. Membrane capacitance was 64.0 +/- 11.5 pF (n = 8). 3. At 0.1 microM, 12-O-tetradecanoylphorbol-13-acetate (TPA) and 4 beta-phorbol-12-13-dibutyrate (PDB) enhanced IK at +80mV by 14.5 +/- 2.0% (n = 8, P < 0.05) and 23.5 +/- 2.2% (n = 7, P < 0.01). Also, ITO at +80mV was increased by 22.1 +/- 2.1% (n = 8, P < 0.01) at 1 microM TPA and by 22.7 +/- 3.0 (n = 7, P < 0.05) at 0.1 microM PDB, significantly. 4. These results indicate that the IK and ITO currents are present in the uterine smooth muscle cells of pregnant rat, and PK-C stimulation modulates the K+ currents, resulting in the regulation of physiological contraction of the uterine muscle during pregnancy. PMID- 8723527 TI - Effect of endothelin on angiotensin converting enzyme activity in cultured vascular smooth muscle cells. AB - We evaluated the effect of endothelin-1 (ET) on the angiotensin converting enzyme (ACE) activity in rat aortic smooth muscle cells (VSMCs). ACE activity was determined by radioimmunoassay of the amount of angiotensin II generated after the addition of angiotensin I (500 pg/ml) to cultured VSMCs. The antibody used had less than 0.1% cross-reactivity with angiotensin I. ACE activity increased 1.9-fold 5 hr after the addition of 10(-6) M ET under serum-free conditions. This stimulatory effect of ET on ACE activity in VSMCs was completely inhibited by 10( 7) M captopril. Results suggested that the ACE present in SMCs is stimulated by ET. PMID- 8723526 TI - Twitch responses dependent on external calcium ions in the mouse diaphragm in a potassium-free bathing solution. AB - We examined the effect of external Ca2+ on twitches of mouse diaphragm under the inhibition of Na(+)-K(+)-ATPase in vitro. The muscle was directly stimulated in presence of d-tubocurarine (dTc). K+ removal potentiated the amplitude of twitches with a transient prior reduction. This potentiation depended on external Ca2+. Membrane potentials decreased after removal of both external K+ and Ca2+ and were restored after an addition of Ca2+. The removal of both ions increased the content of Na+ in tissues and decreased K+. These changes were restored to the levels in the K(+)-free bathing solution by the addition of Ca2+. These results imply that Na+/Ca2+ exchange can support twitch contraction under the inhibition of Na(+)-K(+)-ATPase activity. PMID- 8723528 TI - The effect of N6-cyclohexyladenosine and 5'-N-ethylcarboxamidoadenosine on body temperature in normothermic rabbits. AB - 1. Thermal responses to i.v. administration of N6-cyclohexyladenosine (CHA; 0.15 mg/kg), A1 adenosine receptor agonist, or 5'-N-ethylcarboxamidoadenosine (NECA; 0.15 mg/kg), A2 adenosine receptor agonist, were investigated in normothermic rabbits at an ambient temperature (Ta) of 20.0 +/- 1.0 degrees C. 2. Although both compounds inhibited metabolic heat production, only NECA produced hypothermia. 3. NECA showed strong hypotensive activity. 4. Both compounds produced vasoconstriction of the ear skin vessels and CHA, in addition, slowed down the respiratory rate. 5. The role of A1 or A2 adenosine receptors in the thermoregulatory activity of these compounds is discussed. PMID- 8723529 TI - Effect of gliclazide on the functional response to calcium in diabetic rat heart. AB - 1. The cardiac functional response to extracellular Ca2+ in isolated working hearts was evaluated in streptozotocin-induced diabetic rats treated or not treated with gliclazide. 2. Gliclazide treatment of diabetic rats allowed a partial recovery of the body weight decrease, but not of the hyperglycemia nor insulinopenia. 3. The cardiac mechanical response of diabetic rats was altered, especially at high Ca2+ concentration, and 6-week gliclazide treatment restored the dysfunction close to the control values. 4. The results suggest that gliclazide treatment restores the cardiac function of chronic diabetic rats partly through modulating the Ca2+ metabolism. PMID- 8723530 TI - Cigarette smoke-modified low density lipoprotein impairs endothelium-dependent relaxation in isolated rabbit arteries. AB - 1. We investigated the effects of cigarette smoke-modified low-density lipoprotein (CS-LDL) on endothelium-dependent relaxations in various isolated rabbit arteries, and compared them with those of oxidized LDL (Ox-LDL). 2. In aorta rings, acetylcholine-induced endothelium-dependent relaxations were attenuated by preincubation with CS-LDL and Ox-LDL. Endothelium-independent relaxations induced by sodium nitroprusside were not modified. Similar changes were observed in rings of coronary and basilar arteries. 3. These findings indicate that CS-LDL, as well as Ox-LDL, may decrease the release or activity of endothelium-derived relaxing factor in vascular endothelial cells. PMID- 8723531 TI - Elevated local production of neopterin from alveolar macrophages in patients with internal lung diseases. AB - 1. We measured the neopterin level in the supernatant of cultured alveolar macrophages from patients with interstitial lung diseases (ILD patients) as a marker for the activation of alveolar macrophage. 2. In ILD patients, the supernatant neopterin level (40.1 +/- 7.8 pmol/ml) was significantly higher (P < 0.01) than that in control subjects (10.0 +/- 1.6 pmol/ml). 3. We also found that macrophage-colony stimulating factor (M-CSF) and interleukin-2 (IL-2) augmented neopterin production from alveolar macrophage in both ILD patients (51.6 +/- 10.4 and 60.1 +/- 10.8 pmol/ml, respectively, P < 0.01) and control subjects (28.1 +/- 6.0 and 25.7 +/- 4.9 pmol/ml, respectively). 4. These findings suggest that alveolar macrophages produce neopterin by M-CSF or IL-2. PMID- 8723532 TI - Effects of L-glutamic acid and its agonists on snail neurones. AB - The sensitivities of 22 giant neurone types of an African giant snail (Achatina fulica Ferussac) to threo-beta-hydroxy-L-glutamic acid (threo-L-BHGA), a derivative of L-glutamic acid (L-Glu), applied by brief pneumatic pressure ejection, were examined under current clamp. The 5 neurone types were depolarized by this compound, whereas 2 were hyperpolarized. The 4 neurone types, PON (periodically oscillating neurone), RAPN (right anterior pallial nerve neurone), d-RPLN (dorsal-right parietal large neurone) and RPeNLN (right pedal nerve large neurone) that are excited by threo-L-BHGA and one type, v-LCDN (ventral-left cerebral distinct neurone), inhibited by this compound, were selected to study their pharmacological features in detail. Effects of the stereoisomers of L-Glu and threo-L-BHGA, and mammalian L-Glu receptor agonists, ejected by brief pressure, on the 5 Achatina neurone types were examined under voltage clamp. d RPLN produced an inward current (Iin) by L-Glu and threo-L-BHGA, whereas this neurone type was insensitive to D-Glu and erythro-L-, threo-D- and erythro-D BHGA. This was also excited by AMPA, indicating that the pharmacological features of the L-Glu receptors in this neurone type were similar to those of the mammalian ionotropic AMPA type L-Glu receptors. RAPN produced Iin by L-Glu and threo-L-BHGA. This neurone type was also excited by quisqualic acid and ibotenic acid, indicating that the features of the L-Glu receptors were similar to those of the mammalian metabotropic L-Glu receptors. PON and RPeNLN produced Iin by L Glu and threo-L-BHGA. These neurone types were also excited by quisqualic acid, AMPA and ibotenic acid, indicating that their L-Glu receptors seemed to be in the mixed type, of the two types mentioned. On the other hand, v-LCDN produced an outward current (Iout) by threo-L- and erythro-L-BHGA, but was insensitive to L Glu, indicating that the receptors activated by L-BHGA were not L-Glu receptors. This neurone type was also inhibited by quisqualic acid. PMID- 8723533 TI - Pharmacological characteristics of an outward current produced by beta-hydroxy-L glutamic acid on a snail neurone. AB - An outward current (Iout) was produced by stereoisomers of beta-hydroxy-L glutamic acid (L-BHGA), an L-glutamic acid (L-Glu) derivative, applied by brief pneumatic pressure ejection on an identifiable neurone type, v-LCDN (ventral-left cerebral distinct neurone), of Achatina fulica Ferussac. However, L- and D-Glu were almost ineffective on this neurone type. The pharmacological features of this Iout caused by L-BHGA were elucidated in the present study. According to the dose (pressure duration)-response studies on the L-BHGA stereoisomers that produced the Iout, the effective potency of threo-L-BHGA was approximately similar to that of erythro-L-BHGA. The dose (pressure duration)-response curve of quisqualic acid was shifted towards the left direction from those of threo-and erythro-L-BHGA, suggesting that the binding activity of quisqualic acid to the receptors would be stronger than those of the L-BHGA stereoisomers. GABA, glycine and L-homocysteic acid showed an inward current (Iin) on this neurone type, in contrast to the Iout caused by L-BHGA. beta-Alanine and taurine had absolutely no effect. Therefore, no amino acid inhibitory neurotransmitter candidate was found for this neurone type except for L-BHGA. It was assumed that L-BHGA, in either threo-or erythro-configuration, would be an inhibitory neurotransmitter for this neurone type. Mammalian L-Glu receptor antagonists. D(-)-AP-5, (+/-)-CPP, CNQX and L(+)-AP-3, applied by perfusion, showed no effect on the Iout of v-LCDN caused by threo-L-BHGA, indicating that the features of the inhibitory receptor activated by L-BHGA were much different from those of any type of the mammalian L Glu receptors. Among the inhibitors of ATP-sensitive K+ channel, glipizide significantly inhibited the Iout caused by threo-L-BHGA, whereas tolbutamide did not. Inhibitors of intracellular signal transduction systems, H-7, H-8, H-9, staurosporine, calphostin C, KT5823 and W-7, had no effect on the Iout caused by threo-L-BHGA, suggesting that the receptors activated by threo-L-BHGA would be ionotropic. PMID- 8723534 TI - Morphological and biochemical action of dipyrone on rat placenta. AB - 1. The morphological and biochemical action of dipyrone (N-[2,3-dimethyl-5-oxo-1 phenyl-3-pyrazolin-4-yl]-methylamino methanesulfonate, sodium monohydrate) on the placenta of albino rats was studied by means of karyometry of trophoblastic giant cells and by determinations of DNA, RNA and total protein contents. 2. The animals were treated with a single daily dose of 50 mg/kg body weight during 5 different periods: from the 9th to the 12th, 11th to the 14th, 13th to the 16th, 15th to the 18th or 17th to the 20th day of pregnancy. 3. Karyometric results showed that the nuclear volumes of placental cells in rats treated with dipyrone during the first 3 periods were significantly greater than in control animals and that, closer to term, no differences were observed in this regard. Only the animals treated from the 9th to the 12th day of pregnancy had higher placental contents of DNA, RNA and protein than the corresponding controls. 4. Our results showed that dipyrone had a blocking effect on placental cell division which occurs mainly in the initial steps of placental development. PMID- 8723535 TI - Characterization and autoradiographic localization of [3H] alpha,beta-methylene ATP binding sites in cat urinary bladder. AB - 1. The characteristics and distribution of [3H] alpha,beta-methylene ATP ([3H] alpha,beta-MeATP), a radioligand for P2X-purinoceptors, binding sites in cat urinary bladder detrusor were examined. 2. Saturation analysis revealed that, in cat bladder membrane preparations, only one population of binding sites with high affinity (Kd = 1.8 nM) was present, in contrast to other species where both high and low-affinity binding sites are present. Another feature is that the density of the binding sites in the cat bladder (Bmax = 21.2 pmol/mg protein) is considerably higher (about 2-fold) than the high-affinity binding component in the rat bladder membrane preparations. 3. Displacement experiments with unlabelled purinoceptor ligands indicate that [3H] alpha,beta-MeATP mainly binds to P2X-purinoceptors. The order of binding displacement activity was: alpha,beta methylene ATP, beta,gamma-methylene ATP > 2-methylthioATP > ATP > suramin and L beta,gamma-methylene ATP > > adenosine. 4. Autoradiographic study demonstrated dense specific binding sites of [3H] alpha,beta-MeATP on detrusor smooth muscle of cat bladder. 5. The results of this study are consistent with pharmacological studies for the existence of P2X-purinoceptors in cat bladder. PMID- 8723536 TI - Dantrolene blocks the tonic contraction and calcium influx evoked by K+ in ileal longitudinal smooth muscle. AB - 1. Dantrolene (10(-5)-10(-4) M) reduced the K+ (60 mM) tonic response in accordance with decreased Ca2+ uptake, as determined by La method, more than the phasic in ileal muscle. 2. After saponin-treatment of the fibers, which leaves the Ca2+ storage sites intact, dantrolene had only a slight effect on the caffeine-induced contraction. In Triton-X-100-treated fibers, in which the Ca2+ release sites are destroyed, dantrolene did not affect the Ca(2+)-induced contraction. 3. The results suggest that dantrolene inhibited the high-K(+) induced tonic response mainly by inhibiting Ca2+ influx via voltage-dependent Ca2+ channels in ileum without affecting contractile elements. In contrast, dantrolene had only a slight effect on the intracellular Ca2+ release from caffeine-sensitive release sites. PMID- 8723537 TI - Effects of Androctonus crassicauda scorpion venom on endothelium-dependent and independent vascular responses of rabbit aorta. AB - The effects of Androctonus crassicauda scorpion venom on acetycholine (ACh) induced relaxations and contractions of rabbit thoracic aorta were studied. Endothelium-dependent relaxations induced by ACh in phenylephrine-precontracted arteries were enhanced by the scorpion venom. ACh-induced contractions in endothelium-intact open aortic rings were less than those obtained in denuded preparations (n = 6, P < 0.05, ANOVA). Venom (5, 10 and 30 micrograms/ml) potentiated ACh-induced contractions in intact and denuded segments. In the denuded segments, this potentiation was inhibited by indomethacin (10 microM). Thromboxane synthase inhibitor, BW 149H (100 microM) and thromboxane A2 (TXA2) receptor antagonist, R 68070 (10 microM) partly inhibited venom-induced potentiation. NG-nitro-L-arginine (100 microM) increased venom-potentiated ACh responses in intact arterial segments. Venom increased the basal tone by 25-35% at 30 micrograms/ml. These results suggest that A. crassicauda venom may release a relaxing factor from endothelium and contracting factor from the smooth muscle of rabbit isolated thoracic aorta. The contracting factor may be a cyclooxygenase like product, most likely TXA2. The increase in basal tone by 30 micrograms/ml venom was inhibited by phentolamine (10 microM) and guanethidine (10 microM), indicating a venom-induced release of a neurotransmitter from adrenergic nerve endings. PMID- 8723538 TI - Clonidine-induced rhythmic activity in rabbit anococcygeus muscle. AB - 1. Clonidine 0.5 mM induced an extremely regular rhythmic activity in isolated rabbit anococcygeus muscle. The movements were resistant to tetrodotoxin effect. 2. Prazosin (5 x 10(-8)M-5 x 10(-6)M) and yohimbine (1.5 x 10(-7)M-5 x 10(-4)M) showed no remarkable effect on clonidine-induced rhythmic activity. 3. The clonidine-induced contractions were dependent on extracellular calcium and could be inhibited by the omission of calcium from medium or the introduction of verapamil (IC50 = 1.3 x 10(-7)M) or nifedipine (IC50 = 7.5 x 10(-8)M). 4. Pretreatment of animals with reserpine made the preparations 2800-fold more sensitive to this action of clonidine. 5. It can be concluded from this study that clonidine is able to induce rhythmic activity in rabbit anococcygeus muscle through a mechanism that increases intracellular concentration of Ca++ via membrane calcium channels. PMID- 8723539 TI - Endothelium-independent contractile and relaxant responses to histamine in the rabbit aorta and common carotid, mesenteric, renal, and femoral arteries. AB - 1. The role of the vascular endothelium in the relaxant and contractile responses to histamine of the isolated rabbit aorta; common carotid, mesenteric, renal, and femoral arteries; as well as receptor types mediating these responses were analyzed. 2. Histamine (10(-8) to 10(-4) mol/l) contracted resting rings and caused a further concentration-dependent contraction of rings of the arteries precontracted by phenylephrine. 3. Pyrilamine abolished the contractile response to histamine in resting rings of the arteries, whereas it reversed that response into a concentration-dependent relaxant response in precontracted rings of the arteries. The relaxant effect of histamine was abolished by metiamide, but it was not affected by sotalol and atropine. Moreover, in control experiments, the phenylephrine-induced contractions and acetylcholine-induced relaxations were not changed by pyrilamine and metiamide, respectively. 4. Endothelial removal did not influence the contractile and relaxant responses of the arteries to histamine. 5. These findings indicate that, in the isolated rabbit aorta and common carotid, mesenteric, renal, and femoral arteries, the contractile effect of histamine resulting from the activation of H1 receptors overcomes its relaxant effect resulting from the activation of H2 receptors. The effects of histamine are neither mediated nor modulated by the endothelial cells. PMID- 8723540 TI - Mechanism of the potentiating effect of NH4Cl on vasoconstriction in rat aorta. AB - 1. In medium containing 10 or 20 mM KCl, NH4Cl (10 mM) addition significantly diminished tetraphenylphosphonium bromide (TPP+) uptake in A7r5 vascular smooth muscle cells. 2. Addition of 10 mM NH4Cl significantly potentiated the 45Ca2+ uptake of A7r5 cells in medium containing 20 mM KCl; this increased uptake could be reduced to the basal level by pretreatment with 0.1 microM verapamil. 3. In rat aortic strips, KCl at 10 mM induced a slight contraction, that was greatly potentiated by the simultaneous addition of 10 mM NH4Cl. In this case, also, pretreatment with verapamil (1 microM) completely abolished the NH4Cl potentiating effect. 4. The results suggest that ammonium ion amplifies the K(+) induced contraction of rat aorta by facilitating transmembraneous Ca2+ influx through the voltage-dependent calcium channel, which may be due to potentiation of KCl-induced depolarization of the plasma membrane. PMID- 8723541 TI - Effect of left ventricular pressure and volume overload on alpha-adrenoceptor activity in patients with rheumatic heart valvular disease. AB - We have investigated the possibility that the various left ventricular load conditions may exert different effects on the sympathetic function by comparing the influence of volume (VOL) and pressure (POL) overload on platelet alpha adrenoceptor activity, plasma catecholamines and cAMP in 44 patients with rheumatic heart valvular disease. Receptor activity was determined by radioligand binding methods, catecholamines by HPLC using an electrochemical detector, and cAMP by radioimmunoassay. The mean alpha-adrenoceptor density (Bmax) of the control group (n = 29) was 4.71 +/- 0.41 fmol per 10(7) platelets and the corresponding dissociation constant (Kd) was 2.47 +/- 0.15 nM. In VOL patients, the density was elevated by 70% (P < 0.0001), but it remained unchanged in the POL patients. In contrast to the Bmax, the Kd of the VOL group was not changed, and it increased by 34% (P < 0.01) in the POL group. Norepinephrine was elevated by 91% (P < 0.05) in POL, and epinephrine increased by 65% (P < 0.05) in POL and 71% (P < 0.05) in VOL. These results suggest that the sympathetic nervous system responds to left ventricular volume overload by increasing alpha-adrenoceptor density with no apparent change in receptor affinity toward [3H]-yohimbine binding, and to left ventricular pressure overload by decreasing their binding affinity without a parallel decrease in receptor density. The increase in receptor density in VOL is accompanied by an increase in plasma epinephrine, and the decrease in binding affinity in POL is associated with increased plasma norepinephrine and epinephrine levels. PMID- 8723542 TI - Role of polymorphonuclear leukocytes and oxygen-derived free radicals in chronic gastric lesion induced by acetic acid in rat. AB - This study examined the role of oxygen-derived free radicals, the potential involvement of neutrophils and the possible mucosal vascular permeability changes involved in the pathogenesis and evolution of gastric mucosal lesions induced by acetic acid in the rat. Myeloperoxidase activity was assayed and used as an index of leukocyte infiltration. Application of acetic acid produced a significant increase in this activity 7 and 14 days after induction of chronic injury. Administration of hydroxyurea intraperitoneally was associated with a decrease in the severity of chronic ulceration and neutrophil infiltration into the gastric lesion. This effect was detectable enzymatically and microscopically. Orally administered allopurinol did not produce any beneficial effects on either the macroscopic and histological appearance or on vascular permeability. These results suggest that oxygen-derived free radicals may contribute to the formation and development of chronic lesions and that oxygen-derived free radicals were generated from neutrophils, but not from the xanthine oxidase pathway. These inflammatory cells may, therefore, have a lesive role in the origin and course of acetic acid ulcer disease. PMID- 8723543 TI - Cardiac electrophysiological effects of levosimendan, a new calcium sensitizer. AB - 1. The conventional microelectrode and the patch-clamp techniques were used to study the electrophysiological effects of levosimendan, a new calcium-sensitizing cardiotonic drug, in cardiac ventricular muscle. 2. Levosimendan (5 microM) did not change the main repolarizing currents, such as the inward rectifier potassium, transient outward and the delayed rectifier outward potassium current, in rabbit ventricular myocytes. 3. In rabbit ventricular muscle, levosimendan, at relatively low concentrations (0.1-1 microM), did not change significantly the amplitude of the inward calcium current but increased the amplitude of the twitch tension. 4. In guinea pig ventricular muscle, levosimendan, at higher concentrations (1-5 microM), significantly increased the amplitude of the inward calcium current and the slow-response action potential parameters. 5. It is concluded that levosimendan, in addition to its calcium sensitizing properties characterized by "silent electrophysiology," exhibits cardiac electrophysiological effects similar to those of phosphodies-terase inhibitors. PMID- 8723544 TI - Synthesis of peroxynitrite in a two-phase system using isoamyl nitrite and hydrogen peroxide. AB - A new method for the preparation of high concentrations of peroxynitrite (up to 1 M) is described. The synthesis uses a two-phase system and involves a displacement reaction by the hydroperoxide anion (in the aqueous phase) on isoamyl nitrite (in the organic phase). The product peroxynitrite remains in the aqueous phase, whereas isoamyl alcohol forms a new organic phase along with the unreacted isoamyl nitrite. The aqueous phase contains some 0.15 M isoamyl alcohol and the unreacted hydrogen peroxide, but no isoamyl nitrite. Removal of isoamyl alcohol or traces of isoamyl nitrite is accomplished by washing the aqueous phase with dichloromethane, chloroform, or hexane. A near total removal of hydrogen peroxide is then achieved by passing the solutions through a short column of manganese dioxide. The peroxynitrite in these postprocessed solutions has broad absorption spectrum with a maximum around 302 nm, follows a characteristic first order decomposition at pH 7.2 and 25 degrees C (k = 0.34 +/- 0.1 s-1), and reacts with organic compounds to give either nitrated or one-electron transfer products. When stored frozen at -20 degrees C, these peroxynitrite solutions decompose at a rate of about 1.7 % per day and should be used within 2-4 weeks. For short-term storage of about 1 week or less, these solutions can be stored at refrigerator temperatures (approximately 5 degrees C) where peroxynitrite has a half-life of about 7 days. PMID- 8723545 TI - Interactive distance education: improvisation helps bridge the gap. AB - In this paper, distance learning through the use of interactive duplex video and audio will be described. The major advantage of this technology is the ability to provide live, face-to-face educational access to distant or isolated students. Yet faculty are concerned about the interrelationships between the instructor and the student and among students in the distance education environment. Improvisation techniques force active participation by the students and can be used to bridge the distance. PMID- 8723546 TI - Computer-based patient education: observations on effective communication in the clinical setting. AB - Computer-based patient education materials are becoming more widely used in an attempt to increase the efficacy and efficiency of the delivery of health care. This paper discusses the authors' experiences in designing and implementing interactive multimedia presentations for patient education in an orthopaedic surgery office setting. Content must be chosen carefully to effectively educate without alienating the patient; simple graphics and animation are best for conveying complex medical and surgical concepts. The interface must be simple; both mouse input and touchscreen have proven acceptable and require minimal instruction. Improvements in program design will result in task-oriented evaluation systems that allow the patient to use the information gained to demonstrate understanding. PMID- 8723547 TI - Health education, the health promotion movement, and communication. AB - This paper explores the debate between health education and health promotion. It is argued that each is located in a different philosophical tradition, The Modern and The Post-Modern. The Health Promotion movement can unintentionally undermine the very basis of prevention. It is argued that a skills-based approach is required. Drawing on the work of Habermas, a model for the effective delivery of the public health education message is outlined. PMID- 8723548 TI - Telephone T.A.L.K.: a teaching/assessment tool for medical training. AB - An instrument for teaching and assessing interpersonal communication skills of physicians on the telephone was developed. A conceptual framework based on the acronym T.A.L.K. [Trust, Assert, Listen and K.I.S.S. (Know, Inquire, Solve, and Stroke)] was used for organizing 22 specific behaviors. Faculty gave feedback to residents using T.A.L.K. guidelines while listening to taped telephone calls of the residents T.A.L.K.ing with patients. A five-point scale enables the instrument to be used for skill assessment and program evaluation. PMID- 8723549 TI - Evaluating telephone T.A.L.K. AB - A telephone communication instrument, Telephone T.A.L.K. (Trust, Assert, Listen, and K.I.S.S.--Know, Inquire, Solve, Stroke), was developed for a pediatric residency program for teaching and assessment purposes. Residents had call reviews with faculty as an intervention. Eight residents had pre-intervention calls and 3 call reviews. At post-intervention, Trust and Listening skills showed significant improvement and Assert and K.I.S.S. near significance. Ranking frequencies of items for calls of 30 residents indicated which items to select for future teaching. PMID- 8723551 TI - Endotherapy for chronic pancreatitis. PMID- 8723550 TI - Localization of gastrinomas. PMID- 8723552 TI - Carbon dioxide transport in rats with acute pancreatitis. AB - CONCLUSION: The production, release, and transport of carbon dioxide from tissues to blood are facilitated both systemically and in the gastrointestinal tract in acute pancreatitis. Red blood cells are responsible for the major exchange and transport of this increase in CO2. The existence of arteriovenous shunting within the intestine is associated with tissue ischemia, which may be involved in the etiology of gut barrier failure in acute pancreatitis. BACKGROUND: Hemodynamic alterations in acute pancreatitis have been described, while little is known about CO2 metabolism. METHODS: Carbon dioxide metabolism was evaluated by virtual values of venoarterial CO2 concentration differences in the early phase after sham operation or induction of acute pancreatitis by intraductal injection of 5% sodium taurocholate in rats. RESULTS: In acute pancreatitis, virtual values of the CO2 concentration increased in arterial RBC at 6 and 12 h as well as in caval and portal vein RBC and plasma. Virtual values of the dissolved CO2 concentration were reduced in arterial and portal vein blood. The increment in blood CO2 concentration related to the increase in CO2 tension from arterial to caval or portal vein valves at constant CO2 tension. The total increment in CO2 concentration from arterial to caval or portal vein blood increased. Whole body oxygen extraction increased, whereas gut oxygen extraction decreased in pancreatitis. PMID- 8723553 TI - Pulmonary infiltrations. Sign of severe acute pancreatitis. AB - CONCLUSION: Pleural infiltrates are indicative of severe acute pancreatitis and a negative prognostic parameter for the course of the disease. BACKGROUND: The purpose of this study was to determine the frequency of pulmonary infiltrates in acute pancreatitis and their importance for evaluating the severity of the disease. METHODS: Incidence and localization of pulmonary infiltrates were evaluated in 140 patients with acute pancreatitis. Chest X-ray was obtained within 24 h, and a contrast-enhanced computed tomography within 72 h after admission. Ranson's prognostic parameters were estimated within the first 48 h. RESULTS: Patients with pulmonary infiltrates (n = 36, 26%) had a significantly higher Ranson's score and pancreatic necroses more frequently than patients without this complication, plus a higher mortality rate. PMID- 8723554 TI - Effects of pancreatic duct ligation and aging on acute taurocholate-induced pancreatitis. Experiments in the perfused pancreas in rats. AB - CONCLUSION: When taurocholate was injected into the common bile duct, high ductal pressure due to ligation of the pancreatic duct did not produce more damage in the pancreas of both old rats and young adult rats, and levels of pancreatic enzymes in portal venous effluent were lower in old rats than in younger rats. BACKGROUND: The effects of ligation of the pancreatic duct and aging on acute pancreatitis caused by taurocholate are still unclear. METHODS: Young adult and old male Wistar rats were used. Six hours after ligation of the common bile duct in both the duodenum and liver hilus, rats were killed and the pancreata were perfused. Taurocholate or normal saline was injected retrogradely into the common bile duct. The levels of amylase and lipase in the portal venous effluent were determined as markers of damage to the pancreas. The pancreas was also histologically examined after the perfusion experiments using an Image Analysis System. RESULTS: (1) A nonsignificant elevation of pancreatic enzymes was found in portal venous effluent by the retrograde injection of saline into the common bile duct. Injection of taurocholate caused a marked elevation of enzymes in the effluent for the first 30 min after injection, which then gradually decreased. (2) Basal levels of pancreatic enzymes were significantly higher in the ligation group than in the nonligation group. Injection of saline into the common bile duct had no apparent effect on enzymes in the effluent. In contrast, taurocholate injection into the common bile duct produced a marked increase in enzymes in the portal venous effluent. However, no significant difference was found between the ligation group and the nonligation group. (3) Similar findings were obtained when old rats were used. (4) Although basal levels of enzymes were almost the same in nonligated old and young adults rats, taurocholate injection into the pancreatic duct in old rats resulted in a significant depression of enzymes compared to that in young adult rats. In the ligation group, pancreatic enzymes in the portal venous effluent following taurocholate injection tended to be lower in old rats than in young adult rats. The results were histologically supported in that various degrees of fibrosis were found in the pancreata of old rats. PMID- 8723555 TI - Complement inhibition by soluble complement receptor type 1 fails to moderate cerulein-induced pancreatitis in the rat. AB - CONCLUSION: Cerulein-induced pancreatitis in rats associated with remote liver and lung dysfunction. Soluble complement receptor 1 (sCR1) does not reduce the local or remote injury. Thus complement activation does not moderate cerulein induced pancreatitis or the associated liver and injury. BACKGROUND: The local and remote injury of pancreatitis resembles other inflammatory events that are mediated by complement. This study examines the effect of complement inhibition with sCR1 in cerulein-induced pancreatitis in rats. METHODS: Thirteen Sprague Dawley rats received five hourly subcutaneous doses of cerulein (100 micrograms initially, then 50 micrograms/kg). Six of these animals received hourly i.v. sCR1 (15 mg/kg, a proven complement-inhibiting dose in rats) and the other seven received i.v. saline. In parallel, 12 additional rats received hourly s.c. and i.v. saline. RESULTS: Compared to saline controls, rats receiving cerulein showed increased pancreatic wet-to-dry ratio (3.25:8.52), hematocrit (40 to 47%), ascites volume (2.1 to 6.1 mL), serum amylase (1680 to 10,700 U/L), and ascites amylase (32,200 to 167,000 U/L) (all p < 0.05). None of these parameters were modified by treatment with sCR1. Serum SGPT, which increased from 33.4 to 46.6 U/L in cerulein-infused rats, showed a trend toward reduction to 38.8 U/L in rats treated with sCR1. Cerulein-treated rats also had increased lung myeloperoxidase (0.069 to 0.097 U/g) and lung permeability, as assessed by a alveolar lavage to serum ratio of labeled albumen (0.041:0.121) both p < 0.05). Neither were changed by sCR1 treatment. PMID- 8723556 TI - Granulocyte-colony stimulating factor produced by pancreatic carcinoma. AB - CONCLUSION: A rare case of granulocyte-colony stimulating factor (G-CSF) produced by carcinoma of the pancreas has been reported. BACKGROUND: This is the first case showing high G-CSF concentration in the aspirated tumor fluid (mucin) at its early stage without leukocytosis. METHODS: The tumor, detected incidentally in a 64-yr-old male, was removed by a distal pancreatectomy. The mass was 7.0 x 6.5 x 4.5 cm, and was histologically diagnosed as cystadenocarcinoma with prominent sarcomatous transformation. It was classified as anaplastic carcinoma. RESULTS: After 4 wk of resection, progressive leukocytosis was observed. Seven weeks after the operations, the peripheral leukocyte count increased to 126,000/mL. After 8 wk of resection, the patient died of recurrence. The serum G-CSF concentration was elevated after recurrence. The preserved mucin contained in the cystic components of the resected specimen had a G-CSF concentration higher than 2400 pg/mL. G-CSF is a known cytokine and an etiologic agent in paraneoplastic syndromes. An early diagnosis can, therefore, be made prior to the manifestation of clinical symptoms by the evaluation of the aspirated tumor fluid. This can lead to the prevention of the paraneoplastic syndrome with inhibitory cytokines in future. PMID- 8723557 TI - Serous cystadenoma of the pancreas communicating with a pancreatic duct. AB - CONCLUSION: To differentiate serous cystadenoma from other cystic lesions communicating with the pancreatic duct, careful radiological examination is necessary. BACKGROUND: Communication between the cystic cavity and the pancreatic duct in an ordinary serous cystadenoma is uncommon, although it is not uncommon in other cystic lesions, such as pseudocyst, mucinous cystadenoma/adenocarcinoma, and intraductal papillary tumor. METHODS. A serous cystadenoma of the pancreas communicating with main pancreatic duct in a 76-yr-old male is reported. RESULTS: The communication was preoperatively demonstrated by endoscopic retrograde cholangiopancreatography and confirmed by histopathological examination of the resected specimen. PMID- 8723558 TI - Solitary pancreatic metastasis occurring eight years after nephrectomy for renal cell carcinoma. A case report and surgical review. AB - CONCLUSION: Pancreatic metastasis from renal cell carcinoma is extremely rare. The average time between nephrectomy and the diagnosis of metachronous metastases is reported to exceed 10 yr. Therefore, the initial diagnosis may be neglected in the cases of prolonged disease-free interval. When it does occur simultaneously or metachronously, aggressive surgical resection, when possible, seems to be the most effective treatment for this metastatic lesion. BACKGROUND: An 81-yr-old female patient, who 8 yr previously had undergone right radical nephrectomy for renal cell carcinoma, presented with solitary pancreatic metastasis, which was successfully treated with a distal pancreatectomy. Only 66 cases of clinically diagnosed renal cell carcinoma metastatic to the pancreas are reported in the world literature and 49 of the patients (including ours) underwent a definitive surgical resection. Our case, treated by distal pancreatectomy, and a review of the relevant literature including all reported cases of renal cell carcinoma metastatic to the pancreas, are presented. RESULTS: The patient was well without any evidence of recurrence at 22 mo after the operation. PMID- 8723559 TI - Cataracts, radiculomegaly, septal heart defects and hearing loss in two unrelated adult females with normal intelligence and similar facial appearance: confirmation of a syndrome? AB - Two unrelated, adult females with normal intelligence are described. They show a similar clinical picture with a long and narrow face, congenital cataract, microphthalmia, microcornea, a high nasal bridge, a short nose, a broad nasal tip, a long philtrum, bilateral hearing loss, persistent primary teeth, oligodontia, variable root length including dental radiculomegaly, heart defects and cutaneous syndactyly of the 2nd-3rd toes. Abnormalities present in only one of the two patients were a cleft palate and a transverse vaginal septum, respectively. There are numerous similarities between our two patients and the family described by Wilkie et al. ((1993): Clin Dysmorphol 2: 114-119) and all may be examples of the same entity. PMID- 8723560 TI - Serpentine fibula syndrome: expansion of the phenotype with three affected siblings. AB - We describe three siblings, one of whom has serpentine fibula syndrome (SFS) and has many facial and skeletal features in common with two deceased brothers, making it highly likely that they too had the condition. The karyotype of one of the deceased males was 47,XXY. These are the first affected sibs with SFS, and the first affected males. They all have other abnormalities which have not previously been described as part of the condition, namely congenital heart disease, inguinal herniae (two sibs), intestinal malrotation (two sibs) and coloboma (one sib). Facially they resemble the cases described by ter Haar et al. (1983), who also had congenital heart disease and a skeletal dysplasia though did not have the characteristic bowing of the fibulae. There are also features in common with Hadju-Cheney syndrome. PMID- 8723561 TI - The EEC syndrome: a literature study. AB - Analysis of 230 published cases of the EEC syndrome revealed that, besides the cardinal symptoms (ectrodactyly, ectodermal dysplasia and clefting), lacrimal tract abnormalities and urogenital abnormalities are part of this syndrome. Mental retardation and various abnormalities and dysmorphisms of the ears and face, reported in EEC syndrome, do not really seem to be part of it. Conductive hearing loss is associated with clefting. A score for the severity of symptoms in the EEC syndrome is described, and using this score it appears that isolated cases are generally more severely affected than familial cases. We did not find signs of genomic imprinting or anticipation in published EEC families. We did find, however, that interfamilial variability is significantly larger than intrafamilial variability, pointing to genetic (allelic?) heterogeneity. The penetrance of the EEC-mutation is estimated to be between 93% and 98%. PMID- 8723562 TI - Three sibs with microcephaly, congenital heart disease, lung segmentation defects and unilateral absent kidney: a new recessive multiple congenital anomaly (MCA) syndrome? AB - We present three sibs with similar dysmorphic features, including brain, heart and lung malformations that have not been reported previously. Cytogenetic studies have failed to demonstrate a chromosomal abnormality. Although parental consanguinity was not present, we suggest that this may constitute a new recessive syndrome. PMID- 8723563 TI - Schinzel-Giedion syndrome: further delineation of the phenotype. AB - We describe a male infant with findings typical of Schinzel-Giedion syndrome. Characteristic features include: midface retraction, widely patent fontanelles, hirsutism, choanal stenosis, hypospadias with chordae, club feet and broad ribs. The patient suffered from seizures and died at 14 months of age of fulminant bronchopneumonia. Pathological examination revealed steatosis in the liver as well as lipid vacuolization of the zona fasciculata of the adrenals. PMID- 8723564 TI - PEHO or PEHO-like syndrome? AB - PEHO syndrome is a rare progressive infantile encephalopathy, with variable age of onset of hypotonia, convulsions, mental retardation, oedema, and optic atrophy. Neuroimaging shows cerebellar and brainstem atrophy in most instances. A PEHO-like syndrome has been described in which those affected do not have the typical changes on neuroimaging. We report four new cases, two isolated cases and two sisters, who might be part of the PEHO-like syndrome. PMID- 8723565 TI - An unusual presentation of Smith-Magenis syndrome with iris dysgenesis. AB - A boy who presented with iris dysgenesis is described. He was shown to have Smith Magenis syndrome with a deletion of 17p11.2. PMID- 8723566 TI - A new lethal neonatal short limb dwarfism. AB - We report on a baby with an apparently new type of lethal short limb dwarfism. The inheritance appears to be autosomal recessive. PMID- 8723567 TI - The Jarcho-Levin syndrome (spondylocostal dysplasia) and complex congenital heart disease: a case report. PMID- 8723568 TI - Bilateral brachial amelia, facial clefts, encephalocele, orbital cyst and omphalocele: a recurrent fetal malformation pattern coming into focus. AB - Ultrasonographic and pathoanatomical features of a fetus displaying bilateral brachial amelia, complex facial clefting, encephalocele and omphalocele are described. The findings are compared with the few previously published cases. Bilateral brachial amelia with complex other organ malformations in previable fetuses may be more frequently diagnosed with improving prenatal ultrasonography. PMID- 8723569 TI - A large inguinal hernia with undescended testes and micropenis in Robinow syndrome. AB - We present a 1 month-old infant with Robinow syndrome and large inguinal hernia, undescended testes and micropenis mimicking penile agenesis. Genital anomalies are common in Robinow syndrome. The ambiguous genitalia appearance may lead to confusion in gender assignment. PMID- 8723570 TI - Polysyndactyly and trigonocephaly with partial agenesis of corpus callosum. AB - Several malformation syndromes which include polysyndactyly and craniofacial anomalies have been described. We report a case of an 11-month-old boy with a pattern of anomalies including polysyndactyly, trigonocephaly, partial agenesis of corpus callosum and dysmorphic facies. PMID- 8723571 TI - Another patient with an unusual syndrome of mental retardation and sparse hair? PMID- 8723572 TI - Potential of epoetin alfa in patients in autologous blood donation programs for orthopedic surgery. AB - The ability of epoetin alfa to increase hematopoiesis in a dose-dependent manner when administered by the intravenous (i.v.) or subcutaneous (s.c.) route has been demonstrated in pharmacokinetic studies in healthy volunteers. Epoetin alfa may therefore be a useful adjunct to autologous blood (AB) donation. By facilitating AB donation, the use of allogeneic blood could be reduced. In patients scheduled to undergo orthopedic surgery, i.v. administration of epoetin alfa 600 IU/kg twice weekly for 3 weeks prior to surgery (in conjunction with oral iron supplementation) significantly increased the number of AB units and total red blood cell (RBC) volume donated and increased the number of patients able to donate > or = 4 AB units. However, there was no difference between epoetin alfa and placebo groups with respect to allogeneic blood exposure. PMID- 8723573 TI - Erythropoietin, iron metabolism, and red blood cell production. AB - Erythropoietin (EPO) plays a central role in the regulation of red blood cell (RBC) production. Since iron is an essential element for erythropoiesis and hemoglobin (Hb) synthesis, its importance is heightened in patients treated with epoetin alfa. Stimulation of erythropoiesis following the administration of epoetin alfa is associated with several changes in iron metabolism; indeed, plasma ferritin levels fall as a result of increased utilization of iron by the expanding erythroid marrow. The administration of epoetin alfa can therefore lead to a state of relative iron deficiency. Thus, iron supplementation is essential to maximize the effect of epoetin alfa-induced erythropoiesis. PMID- 8723574 TI - Epoetin alfa increases the volume of autologous blood donated by patients scheduled to undergo orthopedic surgery. AB - In patients scheduled to undergo major orthopedic surgery, predonation of autologous blood (AB) has emerged as a means of avoiding subsequent exposure to allogeneic blood. However, patients with a baseline hematocrit (Hct) less than 40% may not be able to donate sufficient AB to fully meet their requirements. In female patients with a baseline Hct < or = 39%, epoetin alfa (300 to 600 IU/kg twice weekly for 3 weeks) significantly increased the amount of AB donated prior to elective orthopedic surgery and significantly reduced allogeneic blood requirements in comparison with placebo. Iron availability was a critical factor in determining the response to epoetin alfa. In these patients, parenteral supplementation with iron saccharate significantly increased the amount of AB donated and the volume of red blood cells (RBCs) collected in comparison with oral iron alone. Parenteral iron supplementation, therefore, ensures that sufficient iron is available to meet the demands of epoetin alfa-accelerated erythropoiesis in patients enrolled in an AB donation program. PMID- 8723575 TI - Epoetin alfa as an adjunct to autologous blood donation in patients with a low hematocrit scheduled for elective orthopedic surgery. AB - Predonation of autologous blood (AB) represents an attractive alternative to allogeneic blood transfusion in patients scheduled for elective orthopedic surgery. However, anemic patients may not be able to donate sufficient AB prior to surgery, thus increasing the risk of exposure to allogeneic blood. This group of patients may benefit from the administration of epoetin alfa to facilitate AB donation. A recent multicenter, double-blind, placebo-controlled study in 204 patients with a low hematocrit (Hct; < or = 39%) scheduled for orthopedic surgery demonstrated that the intravenous administration of epoetin alfa (600 IU/kg twice weekly for 3 weeks) significantly increased the number of AB units predeposited and the percentage of patients able to donate > or = 4 AB units. Furthermore, epoetin alfa attenuated the decrease in Hct associated with AB donation and significantly (P = .027) reduced allogeneic blood exposure in these patients. PMID- 8723576 TI - Epoetin alfa for autologous blood donation in patients with rheumatoid arthritis and concomitant anemia. AB - In patients scheduled for major orthopedic surgery, the presence of anemia can preclude the donation of sufficient autologous blood (AB) to meet transfusion requirements. Although a number of studies have investigated the use of epoetin alfa (in conjunction with parenteral iron supplementation) to facilitate AB donation and reduce exposure to allogeneic blood in this patient population, the optimum treatment regimen and route of administration has yet to be defined. In rheumatoid arthritis (RA) patients with a low predonation hematocrit (Hct; < or = 39%), intravenous (i.v.) treatment with epoetin alfa 300 IU/kg twice weekly for 3 weeks was the optimum dosage for facilitation of AB donation and minimization of the decrease in Hct prior to elective orthopedic surgery. However, the subcutaneous (s.c.) route of epoetin alfa administration may allow lower dosages of epoetin alfa to be used. Indeed, epoetin alfa 100 IU/kg s.c. twice weekly for 3 weeks (in conjunction with a single i.v. bolus of 200 IU/ kg at the first s.c. dose) was as effective as 300 IU/kg i.v. administered according to the same schedule. The number of AB units collected, total red blood cell (RBC) volume donated, and peak proportion of reticulocytes were similar regardless of the route of administration. Both treatment groups were associated with a significant reduction in allogeneic blood exposure compared with historical controls. Findings consistent to all of these studies were that epoetin alfa was well tolerated, and that i.v. iron supplementation was necessary to maximize its beneficial effects. PMID- 8723577 TI - Epoetin alfa plus autologous blood donation in patients with a low hematocrit scheduled to undergo orthopedic surgery. AB - A low predonation hematocrit (Hct) can preclude the collection of sufficient autologous blood (AB) to meet the transfusion requirements of patients scheduled for orthopedic surgery. Subcutaneous (s.c.) administration of epoetin alfa, in conjunction with intravenous (i.v.) iron supplementation, has proved effective for the facilitation of AB donation by such patients. Compared with untreated controls and patients treated with i.v. iron alone, epoetin alfa 50 to 150 IU/kg SC plus i.v. iron twice weekly for 3 weeks prior to surgery significantly increased total red blood cell (RBC) production (P < .01) and the volume of RBCs donated (P < .05). Epoetin alfa was particularly effective in females and patients with a predicted blood volume (PBV) less than 5 L. Treatment with epoetin alfa led to an increase (albeit nonsignificant) in the number of AB units predonated compared with i.v. iron alone. However, in patients with a PBV less than 5 L, a substantially greater percentage of epoetin alfa-treated patients donated > or = 4 AB units (80% v 30%). Allogeneic blood requirements were reduced, albeit not significantly (P = .051), in patients treated with epoetin alfa. However, in comparison with untreated controls, there was a significant reduction in the mean volume of allogeneic blood transfused per transfused patient in the epoetin alfa groups. The optimum s.c. dose of epoetin alfa in patients with a low predonation Hct scheduled for orthopedic surgery appears to be between 100 and 150 IU/kg twice weekly for 3 weeks. PMID- 8723578 TI - Epoetin alfa as an adjuvant to autologous blood donation. AB - The development of anemia is one factor that can limit the donation of sufficient autologous blood (AB) to meet a patient's expected blood requirements following elective orthopedic surgery. Two clinical studies have been conducted in nonanemic patients to investigate the use of epoetin alfa as an adjuvant to AB predonation to facilitate AB procurement and minimize the development of anemia. Both studies demonstrated that patients with a normal hematocrit (Hct) can donate > or = 3 units of AB prior to surgery. However, treatment with epoetin alfa minimized the decrease in Hct associated with AB donation. While there was a trend toward a reduction in allogeneic blood exposure in patients treated with epoetin alfa, the difference relative to placebo was not significant. This observation may be explained by a limited requirement for blood in this patient population that was met by predonation of 3 AB units. Thus, the use of epoetin alfa as an adjunct to AB predonation is likely to be of most benefit in patients with a normal Hct scheduled for surgical procedures where the expected blood requirements exceed 3 units. In addition, epoetin alfa may enable patients with a low Hct, low body weight, or low predicted blood volume to participate in or to complete an AB donation program, thus reducing the possibility of exposure to allogeneic blood. PMID- 8723579 TI - Autologous blood donation plus epoetin alfa in nonanemic orthopedic surgery patients. AB - Intravenous (i.v.) administration of epoetin alfa facilitates the collection of autologous blood (AB) prior to elective orthopedic surgery. However, the optimum dose and dosage regimen remains to be defined. The aim of this multicenter, randomized, placebo-controlled study was to determine the optimum i.v. dose of epoetin alfa (300 IU/kg or 600 IU/kg) that would allow nonanemic patients to donate > or = 4 units of AB within 14 to 21 days of elective orthopedic surgery. All patients (n = 103) received oral iron supplementation and were treated with epoetin alfa or placebo three times during 1 week, within 3 weeks of surgery. Eighty patients were evaluable in the efficacy analysis. Compared with placebo, significantly more evaluable patients treated with epoetin alfa were able to donate > or = 4 AB units. Furthermore, epoetin alfa dose-dependently increased the preoperative reticulocyte count and attenuated the decrease in hematocrit associated with AB predonation. There was no significant difference with respect to allogeneic blood exposure between the epoetin alfa and placebo treatment groups. In addition, the 600 IU/kg dose was not significantly more effective than the 300 IU/kg dose for most efficacy parameters assessed. Several patients became iron-deficient, suggesting that oral supplementation is not an adequate source of iron in these patients. Epoetin alfa 300 IU/kg therefore appears to be the optimum i.v. dose for facilitating the collection of > or = 4 units of AB within 14 to 21 days of elective orthopedic surgery. PMID- 8723580 TI - Epoetin alfa plus autologous blood donation and normovolemic hemodilution in patients scheduled for orthopedic or vascular surgery. AB - In previous studies, treatment with epoetin alfa facilitated preoperative donation of autologous blood (AB). However, some patients may not be able to donate sufficient AB to meet their surgical blood requirements when the time to surgery is short. In this multicenter, double-blind, placebo-controlled study, the ability of epoetin alfa combined with normovolemic hemodilution (NVHD) to facilitate the collection of > or = 4 AB units within 2 weeks of surgery was investigated in 112 nonanemic patients scheduled for elective orthopedic or vascular surgery. All patients received oral iron supplementation and were treated with three intravenous (i.v.) injections of epoetin alfa (300 or 600 IU/kg) on days 1,4, and 7; surgery, in combination with NVHD, was performed on day 13. A total of 4 units of AB were predeposited if the patient's hemoglobin (Hb) level exceeded 11 g/dL at each donation. Compared with placebo, epoetin alfa dose-dependently increased reticulocyte counts prior to surgery and significantly minimized the decrease in hematocrit (Hct) associated with AB donation, although there were no significant differences between dosages. While significantly more patients treated with epoetin alfa were able to donate > or = 4 AB units compared with placebo, there was no difference between the groups in exposure to allogeneic blood. This effect of epoetin alfa was particularly apparent in female patients. I.v. epoetin alfa 300 IU/kg, administered three times within 1 week, appears to be the optimum dose for facilitating the collection of > or = 4 units of AB in nonanemic patients scheduled for elective surgery and NVHD within 2 weeks. PMID- 8723581 TI - Enhanced efficacy of autologous blood donation with epoetin alfa. AB - In the setting of elective orthopedic surgery, epoetin alfa has been shown to facilitate autologous blood (AB) donation and reduce allogeneic blood requirements. However, the most appropriate dose and route of administration remains to be defined. A randomized, double-blind, placebo-controlled study was therefore conducted to compare the effect of different subcutaneous (s.c.) doses of epoetin alfa on hemoglobin (Hb) and endogenous erythropoietin (EPO) levels and transfusion requirements in 62 patients donating AB prior to hip surgery. Epoetin alfa (100 or 200 IU/kg s.c. on days -30, -23, -16, and -9 prior to surgery, in conjunction with oral iron supplementation) dose-dependently ameliorated the reduction in Hb levels associated with AB donation. No patient treated with epoetin alfa required allogeneic blood compared with two placebo recipients (one of whom required additional blood because of reoperation). The results of this study confirm that relatively low dosages of epoetin alfa minimize the decrease in Hb levels during AB donation and help to reduce exposure to allogeneic blood in patients scheduled for elective orthopedic surgery. PMID- 8723582 TI - Autologous blood donation and epoetin alfa in cancer surgery. AB - Patients undergoing cancer surgery frequently require blood, and the transfusion of allogeneic blood in these patients has been controversially linked to an increased risk of tumor recurrence. This patient population may therefore benefit from preoperative autologous blood donation (ABD) with or without epoetin alfa therapy, although the precise impact of autologous blood transfusion has not been fully explored. In some trials, preoperative ABD reduced allogeneic blood exposure by 50% in patients undergoing surgery for cancer resection, while, in another study, perioperative treatment with epoetin alfa significantly increased hematocrit (Hct) levels preoperatively and led to a reduction in postoperative allogeneic blood exposure. A combination of epoetin alfa and preoperative ABD seems a reasonable approach to reducing allogeneic blood exposure in patients undergoing cancer surgery. PMID- 8723583 TI - Effectiveness of perioperative epoetin alfa in patients scheduled for elective hip surgery. AB - Several strategies have been investigated as a means of reducing allogeneic blood requirements in patients undergoing surgery, including the perioperative administration of epoetin alfa. In a multicenter, double-blind, placebo controlled study in 208 patients undergoing elective hip replacement surgery, subcutaneous administration of epoetin alfa (300 IU/kg daily) for 14 or 9 days perioperatively (commencing 10 and 5 days preoperatively, respectively) significantly reduced the incidence of primary outcome events (any allogeneic blood transfusion or a postoperative hemoglobin [Hb] level < 8.0 g/dL) compared with placebo (P = .003). Furthermore, the transfusion requirements of epoetin alfa-treated patients were significantly lower than those of patients treated with placebo (P = .007). Preoperative and postoperative Hb levels and reticulocyte counts were higher in epoetin alfa-treated patients compared with placebo. Epoetin alfa was well tolerated, and the incidence of deep vein thrombosis (DVT) was not different from that observed in placebo recipients. Thus, perioperative administration of epoetin alfa reduces the allogeneic blood requirements of patients undergoing elective hip replacement surgery and is of particular benefit in the subgroup of patients whose baseline Hb levels are less than 13.5 g/dL. PMID- 8723584 TI - Perisurgical use of epoetin alfa in orthopedic surgery patients. AB - In order to avoid exposure to allogeneic blood, perisurgical administration of epoetin alfa has been proposed as an alternative to autologous blood (AB) predonation in patients who are unable or unwilling to donate AB prior to elective surgery. The efficacy of perisurgical epoetin alfa to reduce allogeneic blood exposure was investigated in a randomized, double-blind, placebo-controlled study in 200 patients unable to participate in an AB predonation program who were scheduled for orthopedic surgery with expected blood loss > or = 2 units. Epoetin alfa (100 IU/kg or 300 IU/kg) was administered daily by subcutaneous (s.c.) injection, commencing 10 days preoperatively and continuing until day 4 postoperatively (15 doses in total). All patients received oral iron supplementation. Patients treated with epoetin alfa required significantly fewer (P < .001) allogeneic transfusion compared with placebo, and this effect of epoetin alfa was particularly evident in the subgroup of patients with baseline hemoglobin (Hb) levels of more than 10 to < or = 13 g/dL. In terms of the reduction in allogeneic blood exposure, no significant difference was evident between epoetin alfa regimens. Epoetin alfa was well tolerated. Although 15 s.c. doses of epoetin alfa 100 IU/kg appears to be the optimum dosage regimen in patients scheduled for orthopedic surgery, a presurgical simulation study in 24 healthy volunteers suggested that two s.c. doses of epoetin alfa 600 IU/kg in 10 days prior to expected surgery may be a suitable regimen for further study. However, the optimum timing of epoetin alfa administration in relation to surgery remains to be established. A finding consistent to all studies is that adequate iron supplementation (most probably in parenteral form) is necessary to optimize the erythropoietic response to epoetin alfa in the surgical setting. PMID- 8723585 TI - Epoetin alfa: new directions in orthopedic surgery. AB - The introduction of autologous blood (AB) donation programs has led to a decrease in the number of orthopedic surgery patients exposed to allogeneic blood, although there is still room for improvement. For example, some patients may not be able to donate sufficient AB to meet their expected blood requirements. Virtually all nonanemic patients can donate 3 AB units prior to orthopedic surgery before further AB donation is limited by the development of anemia. In preliminary studies, the administration of epoetin alfa (150 IU/kg subcutaneously (s.c.) on alternate days; six doses) following the donation of 3 AB units reversed phlebotomy-induced anemia and enabled a further 2 units of AB to be collected. The ability of this therapeutic approach to increase AB procurement and reduce allogeneic blood requirements is being investigated in an ongoing, placebo-controlled study. An alternative approach may be to combine perisurgical treatment with epoetin alfa and normovolemic hemodilution (NVHD) prior to orthopedic surgery. Although such studies have yet to be initiated, they may demonstrate a reduction in allogeneic blood exposure in patients unable to donate AB prior to orthopedic surgery, a group of patients traditionally at high risk of exposure to allogeneic blood. PMID- 8723586 TI - Autologous blood donation with recombinant human erythropoietin in cardiac surgery: the Japanese experience. AB - Four units of predonated autologous blood (AB) is considered sufficient to cover the blood requirements of 95% of patients undergoing elective cardiac surgery, thus avoiding the risks associated with allogeneic blood transfusion. A review of six Japanese studies was undertaken to summarize the potential for recombinant human erythropoietin (rHuEPO) to facilitate donation of AB by patients scheduled for cardiac surgery. Intravenous (i.v.) administration of rHuEPO improved the anemia associated with AB donation, an effect that was further enhanced by i.v. iron supplementation. Once weekly subcutaneous (s.c.) administration of rHuEPO facilitated the donation of AB and reduced allogeneic blood requirements in patients scheduled for cardiac surgery, suggesting that rHuEPO could be administered on an outpatient basis. rHuEPO was of particular benefit in anemic patients, eliminating exposure to allogeneic blood in the majority of patients. In conclusion, rHuEPO facilitates the donation of AB and reduces allogeneic blood requirements of patients scheduled for cardiac surgery. PMID- 8723587 TI - Subcutaneous epoetin alfa as an adjunct to autologous blood donation before elective coronary artery bypass graft surgery. AB - Autologous blood (AB) donation can minimize exposure to allogeneic blood in patients scheduled for coronary artery bypass graft (CABG) surgery. During AB donation in this group of patients, minimization of the accompanying decrease in hemoglobin (Hb) levels is important to reduce the risk of provoking silent myocardial ischemia and/or arrhythmias. Recombinant human erythropoietin (rHuEPO) has been used to facilitate AB donation and minimize the accompanying decrease in Hb levels in patients scheduled for cardiac surgery. In 24 patients scheduled for CABG surgery, once-weekly subcutaneous (s.c.) administration of rHuEPO (epoetin alfa 400 IU/kg) plus oral iron supplementation for 4 weeks prior to surgery caused marked stimulation of erythropoiesis and significantly increased collection of autologous red blood cells (RBCs) compared with oral iron alone. Furthermore, epoetin alfa minimized the decrease in Hb levels associated with AB donation and significantly attenuated allogeneic blood requirements by facilitating the collection of 4 AB units prior to surgery. During AB donation, no changes in the incidence or severity of ischemic attacks or ST-segment changes were observed using electrocardiographic monitoring. Epoetin alfa was well tolerated. Once-weekly s.c. administration of epoetin alfa for 4 weeks therefore represents a practical means of facilitating AB donation by patients scheduled for CABG surgery. PMID- 8723588 TI - Use of epoetin alfa in autologous blood donation programs for patients scheduled for elective cardiac surgery. AB - The optimum dosage of subcutaneous (s.c.) epoetin alfa was assessed in a double blind study in 31 patients scheduled for cardiac surgery. Patients received a total of four doses of either epoetin alfa 150 IU/kg (n = 11), epoetin alfa 300 IU/kg (n = 10), or placebo (n = 10) administered as single s.c. injections at weekly intervals starting 23 days prior to surgery. AB was collected with isovolemic replacement prior to each of the first three doses of medication. During the AB donation period, Hb levels decreased significantly (P < .05) from baseline to surgery in the placebo group (16.5%), compared with no significant decrease in either of the epoetin alfa groups (8.1% and 9.7% in the 150 IU/kg and 300 IU/kg groups, respectively). In addition, the difference between groups with regard to the decrease in Hb level reached statistical significance (P < .05) for the 150 IU/kg group versus placebo. Epoetin alfa treatment was also associated with significantly higher reticulocyte counts and serum erythropoietin levels in the preoperative period compared with placebo. PMID- 8723589 TI - Consensus statement: using epoetin alfa to decrease the risk of allogeneic blood transfusion in the surgical setting. Roundtable of Experts in Surgery Blood Management. PMID- 8723590 TI - QT interval in cardiac amyloidosis. PMID- 8723591 TI - The calcium antagonist scare in America: what have I learned? PMID- 8723592 TI - Restenosis--an open file. AB - The main procedural drawback to percutaneous coronary angioplasty is restenosis of the treated site within 6 months. Despite advances in equipment, technique, and adjunctive therapies, restenosis has occurred in approximately one-third to one-half of all patients. The biology of restenosis can be divided into plaque persistence and recoil, thrombus formation and transformation, and cellular proliferation and vascular remodeling. Animal models of restenosis have helped to elucidate these mechanisms of restenosis and provide a means to test pharmacologic and mechanical strategies to reduce stenosis recurrence. While numerous agents have been tested in animal models, until recently none has translated into benefit in large-scale clinical trials. Two therapeutic "hopefuls" which have recently emerged in clinical practice are the potent platelet inhibitors, glycoprotein IIb/IIIa receptor antagonists, and intracoronary metallic stents. The IIb/IIIa receptor antagonists target thrombus formation at the angioplasty site, thereby minimizing abrupt vessel closure acutely and neointimal growth chronically, while intracoronary stents safely produce a large coronary arterial lumen acutely and prevent vessel recoil. Separately, these therapeutic strategies have been shown to reduce clinical restenosis 20-30% at 6-month follow-up. With these encouraging results, the future will certainly provide more pharmacologic and mechanical therapies targeting restenosis. With increased understanding of the restenotic process and continued refinement of effective treatments, it may be possible one day to prevent stenosis recurrence. PMID- 8723593 TI - Antiplatelet and anticoagulant use after myocardial infarction. AB - Coronary thrombosis leading to myocardial infarction is a complex process involving the interaction of the arterial wall, the coagulation cascade, and platelets. Increased understanding of the molecular biology of thrombosis has prompted an evolution in antithrombotic therapy, from the early use of warfarin following myocardial infarction to agents targeting specific receptors or modulators in the thrombotic process. The complexity of thrombosis allows for numerous sites of pharmacologic intervention; the multiple pathways leading to platelet aggregation and thrombin formation provide the opportunity for combined therapies. This review presents the current clinical data on antiplatelet, anticoagulant, and specific antithrombin therapies following myocardial infarction. PMID- 8723594 TI - Clinical importance of viability assessment in chronic ischemic heart failure. AB - BACKGROUND AND HYPOTHESIS: Revascularization has provided an effective treatment of depressed left ventricular function in patients with chronically ischemic or "viable" myocardium. Assessment of viable myocardium can be achieved by several noninvasive techniques including dobutamine stress echo or radionuclides such as flurodeoxyglucose (F18DG). F18DG uptake studies are based on the assumption that enhanced glucose uptake in areas of diminished blood flow provides evidence of viable myocardium. To determine the clinical utility of viability assessment in the management of chronic ischemic left ventricular dysfunction, we reviewed the findings and short-term treatment of a series of patients referred for heart failure evaluation who had subsequent F18DG uptake scans. METHODS: We retrospectively reviewed 59 consecutive F18DG viability studies in a series of patients who had documented coronary artery disease and depressed left ventricular function. Single photon emission computerized tomography (SPECT) with F18DG was performed in the patients and these images were compared to SPECT images of resting myocardial perfusion using thallium, sestamibi, or teboroxime. Clinical decisions based on the results of these scans were obtained from chart review. Thirty-day mortality was determined from chart review or contact with the patient's physician. The patients were divided into those without and with F18DG uptake consistent with viable ischemic myocardium. Further analysis included subgroups of patients who were advised to undergo transplantation, revascularization, or to continue medical therapy. RESULTS: Of 34 patients referred for cardiac transplantation, 18 had viable myocardium and 13 underwent revascularization. In the entire study group, 34 of 59 (58%) had evidence of viable myocardium and 29 had subsequent revascularization procedures. Thirty-day survival for all revascularization patients was 86%. CONCLUSION: Assessment of myocardial viability with F18DG SPECT imaging in patients with ischemic left ventricular dysfunction led to a clinical decision for revascularization in approximately half the patients with severe coronary disease and left ventricular dysfunction who were evaluated for myocardial viability in our institution. PMID- 8723595 TI - Symptoms of hypertrophic cardiomyopathy, with special emphasis on syncope and postprandial exacerbation of symptoms. AB - We undertook a prospective study of the symptoms of hypertrophic cardiomyopathy with the aim of profiling symptomatic morbidity in detail, determining the prevalence of anxiety and depression, and describing the prevalence and associations of syncope and postprandial symptom exacerbation. A questionnaire was administered to consecutive outpatients; 70 with hypertrophic cardiomyopathy, 43 with coronary artery disease, 32 with idiopathic dilated cardiomyopathy, and to 40 normal subjects. Hypertrophic cardiomyopathy patients underwent exercise testing, echocardiography, and Holter monitoring. Hypertrophic cardiomyopathy patients had a high frequency of cardiac symptoms and, on average, had a level of symptomatic morbidity equivalent to that of chronic stable angina and dilated cardiomyopathy. There was no evidence for an excess of anxiety (14%) or depression (6%) in patients with hypertrophic cardiomyopathy. Syncope and presyncope, especially provoked by exertion or posture change, were characteristic and common symptoms in hypertrophic cardiomyopathy. A history of syncope was associated with an abnormal blood pressure response to exercise in over 50% of cases that may be the mechanism of syncope in some. Postprandial exacerbation of symptoms occurred in over one-third of hypertrophic cardiomyopathy patients, half of coronary disease patients, and infrequently in dilated cardiomyopathy. Hypertrophic cardiomyopathy patients with postprandial symptoms had a greater frequency of angina, were more symptomatic, and had a reduced exercise capacity, suggesting that postprandial symptoms are a marker for more severe disease. PMID- 8723596 TI - Relationship between regional abnormality of left ventricular rapid filling and coronary microcirculation disturbance in hypertrophic cardiomyopathy. AB - BACKGROUND AND HYPOTHESIS: To investigate the mechanism of regional left ventricular diastolic filling disturbance in hypertrophic cardiomyopathy (HCM), we assessed the relationship between abnormalities of regional ventricular rapid filling and regional coronary microcirculation using radionuclide ventriculography and exercise 201thallium (201TI) myocardial scintigraphy with sector analysis. METHODS: Thirty patients with HCM and 14 patients with atypical chest pain syndrome (controls) were studied. Left ventricular images (left anterior oblique view) were obtained by electrocardiogram-gated 99mtechnetium radionuclide angiography and were divided into three sectors radiating from the geometric center. The time-activity curves and their first derivative curves were analyzed, and the peak filling rate (PFR), the ratio of the time-to-peak filling rate per diastolic interval (TPFR/T) were calculated for the global left ventricle and for the lateral and septal sectors. Exercise stress 201TI myocardial scintigraphy was also performed, and early and delayed images were obtained. The regional washout rate (WR) was then evaluated for the lateral and septal sectors. RESULTS: In HCM patients, the regional PFR in the septal sector (corresponding to the region of hypertrophic myocardium) was 285 +/- 76%/s, and was significantly lower than that in the controls (398 +/- 90%/s, p < 0.01). The regional TPFR/T in the septal sector (32 +/- 10%) was prolonged compared with the value of 21 +/- 5% in the controls (p < 0.05). The regional WR in the septal sector was 21 +/- 9%, and was significantly lower than that in the controls (43 +/- 5%, p < 0.01). Moreover, regional WR correlated positively with regional PFR (r = 0.5, p < 0.05) and showed a weak negative relationship with regional TPFR/T (r = -0.4, p < 0.07) in the septal sector. CONCLUSIONS: These results suggest that regional impairment of rapid filling might be related to a disturbance of the coronary microcirculation in HCM. PMID- 8723598 TI - Angioplasty balloon compliance: can in vivo size be predicted from in vitro pressure profile measurements? AB - BACKGROUND AND HYPOTHESIS: This study was undertaken to determine whether the behavior of angioplasty balloons within coronary arteries may differ from that anticipated from data provided by the manufacturers. In particular, the in vitro pressure-diameter profiles may not truly represent in vivo sizes. METHODS: Thus, we assessed the degree of correlation of in vitro with in vivo measurements obtained during routine angioplasty practice. In vivo size of 2.5 mm compliant (n = 8) and 3 mm semicompliant (n = 8) balloons was assessed using quantitative angiography for first, second, and third inflations. RESULTS: In vivo size was less than expected from in vitro measurements. In general balloon diameter increased with inflation pressures up to 8 atmospheres, and some degree of elastic recoil was evident with both balloon types after the last inflation. CONCLUSION: In vivo balloon size may not be accurately predicted from manufacturers' published data. Size is more likely to be affected by factors such as lesion characteristics and elasticity of the vessel wall than by balloon material compliance characteristics. PMID- 8723597 TI - Acute hemodynamic deterioration during rapid atrial pacing in patients with hypertrophic cardiomyopathy. AB - BACKGROUND AND HYPOTHESIS: Supraventricular tachycardia and ventricular tachycardia are often observed in patients with hypertrophic cardiomyopathy (HCM) and they often alter the clinical features of HCM. We examine the influence of supraventricular tachycardia on cardiac function and assess the clinical characteristics of patients with HCM. METHODS: We studied 32 patients with HCM and 8 normal volunteers using echocardiography under transesophageal rapid atrial pacing. RESULTS: Presyncope-associated hypotension was observed during rapid atrial pacing in 8 HCM patients, but in none of the normal controls. During rapid atrial pacing (144 +/- 8 beats/min in HCM, 146 +/- 5 beats/min in controls), systolic blood pressure (SBP), the product of left ventricular filling volume (FV) and heart rate, and fractional shortening (%FS) in the HCM patients decreased significantly compared with the basal values (138 +/- 19 mmHg vs. 99 +/ 24 mmHg, 5.0 +/- 1.2 l/min vs. 2.9 +/- 0.9 l/min, 41.7 +/- 6.2% vs. 35.2 +/- 6.0%, respectively), but these decreases were not observed in normal controls. The decrement of SBP during rapid atrial pacing in HCM patients with a history of syncope was more marked than that in those without such history. The decrement correlated positively with the indices of left ventricular hypertrophy (maximal wall thickness and wall thickness index) and with %FS, and correlated negatively with the endsystolic left ventricular diameter at rest. CONCLUSIONS: In some patients with HCM, supraventricular tachycardia causes marked hemodynamic deterioration that may be related to a history of syncope, marked hypertrophy, hyperkinesis, small cavity size, and small filling volume of the left ventricle. PMID- 8723599 TI - Time to dP/dt(max), a useful index for evaluation of contractility in the catheterization laboratory. AB - HYPOTHESIS: The time from onset of contraction to dP/dt(max), td, is suggested as an index of contractility in the catheterization laboratory. METHODS: We studied 22 normal patients and 18 patients with myocardial failure in the catheterization laboratory. The two groups were completely separated on the td-heart rate (HR) plane. In the normal patients, HR = 73 +/- 19 beats/min, td = 73 +/- 11 ms, and an inverse linear relation td = 109-0.49 x HR (p < 0.001) exist. In the patients with myocardial failure, despite significantly higher HR than in normal patients (HR = 93 +/- 14 beats/min) (p < 0.001), td paradoxically increased (td = 89 +/- 11 ms, p < 0.0001). CONCLUSIONS: These findings support a mathematical analysis of the left ventricular pressure (LVP) during isovolumic contraction in the time domain which shows that td and (dP/dt)/P reflect the time-dependent aspects of contraction and, hence, decrease with increasing contractility. This study shows that td, at any given HR, is a reliable index of contractility. Thus, a ready-to use td-HR plot containing a well-based separation line can provide a reliable and simple method for determining contractility in the catheterization laboratory by examining whether a patient's td value at any HR is below (normal) or above (impaired contractility) the separation line. PMID- 8723600 TI - Acute infarction of the left ventricular papillary muscle: electrocardiographic pattern and recognition of its location. AB - BACKGROUND AND HYPOTHESIS: ST-segment depression during acute myocardial infarction (AMI) is known to herald serious hemodynamic complications. Since the mechanism of this dependence is not clear, we reinvestigated the old concept of papillary muscle infarction (PMI) as a cause of marked ST depression. METHODS: Autopsies and morpho-electrocardiographic cor-relations were performed in 53 patients with AMI involving one or both left ventricular papillary muscles, and in 10 patients with AMI, but without acute PMI. RESULTS: ST-segment depression > or = 1 mm in at least two leads (mean 3.6 +/- 2.2 mm) was found in 46 (86.8%) patients with, and in one without acute PMI. Thus, the sensitivity and specificity in selecting patients with acute PMI from among those with AMI were 86.8 and 90%, respectively, with an overall accuracy of diagnosis of acute PMI in the course of AMI of 87.3%. Among 26 patients with ST elevation consistent with diagnosis of AMI, ST depression, recorded in 22 patients, was insignificantly greater than in 24 of 27 patients without ST elevation: 4.1 +/- 2.9 versus 3.1 +/ 1.2 mm. Localization of ST depression in the limb leads allowed recognition of which papillary muscle suffered from acute infarction: ST depression in the inferior leads was seen only in patients with anterolateral PMI, whereas in leads I and/or a VL it was seen only in cases with posteromedial PMI. This rule was also valid in patients without concomitant ST elevation. CONCLUSION: Patients with acute PMI show marked ST-segment depression. Its location in the limb leads allows recognition of which papillary muscle has undergone necrosis. This cause of marked ST depression in patients with AMI may explain the high mortality in this particular group. PMID- 8723601 TI - Intravenous propafenone in paroxysmal atrial fibrillation: a randomized, placebo controlled, double-blind, multicenter clinical trial. Paroxysmal Atrial Fibrillation Italian Trial (PAFIT)-2 Investigators. AB - BACKGROUND: Pharmacological conversion of paroxysmal atrial fibrillation is frequently necessary. The aim of this study was to compare intravenous propafenone, a class Ic antiarrhythmic agent, with placebo in paroxysmal atrial fibrillation (AF) of recent onset (< 72 h). PATIENTS AND METHODS: We randomly allocated 75 patients, aged 18 to 70 years, with paroxysmal AF to receive intravenous propafenone (2 mg/kg in 15 min followed by 1 mg/kg in 2 h) or the matching placebo. Patients were followed for 3 h. Exclusion criteria were the presence of one of the following: clinical heart failure, recent acute myocardial infarction, hypotension, atrioventricular block, Wolff-Parkinson-White syndrome, or current treatment with antiarrhythmic agents or digitalis. RESULTS: No sign of heart disease was found in 74.7% of the patients. Echocardiographically determined left atrium diameter was similar in the two groups. Conversion to sinus rhythm occurred in 24 of 41 patients allocated to propafenone and in 10 of 34 patients allocated to placebo (odds ratio 3.2, 95% confidence intervals 1.3 7.9; p < 0.01). Mean conversion time was 34 +/- 29 and 71 +/- 55 min, respectively, for propafenone and placebo. Mean heart rate in nonconverters decreased from 146 to 109 beats/min in patients treated with propafenone while it remained virtually unchanged in those treated with placebo. Only minor side effects were noted. CONCLUSIONS: Intravenous propafenone is an effective therapeutic option for restoring sinus rhythm in patients with paroxysmal AF of recent onset. PMID- 8723602 TI - Correlation between left ventricular contractility and relaxation in patients with idiopathic dilated cardiomyopathy. AB - BACKGROUND AND HYPOTHESIS: It is not fully understood whether alterations in left ventricular (LV) relaxation in patients with idiopathic dilated cardiomyopathy (DCM) precede or follow alterations in LV systolic function. Accordingly, we investigated the relationship between LV systole and diastolic relaxation in patients with DCM using LV pressure-volume analysis. METHODS: Hemodynamic data were collected for 38 consecutive patients with DCM in normal sinus rhythm and in the resting steady state (New York Heart Association functional class II to III). Continuous simultaneous pressure-volume relationships were obtained by conductance catheter. Left ventricular end-systolic elastance (Ees) was determined during transient occlusion of the inferior vena cava. RESULTS: Left ventricular ejection fraction, peak negative dP/dt, and peak positive dP/dt were depressed (31 +/- 11%, -1242 +/- 351 mmHg/s, and 1118 +/- 253 mmHg/s, respectively). The LV end-diastolic volume index, the time constants of isovolumic LV pressure decay TL and TD were increased (141 +/- 38 ml/m2, 54 +/- 14 ms, and 69 +/- 17 ms, respectively). Ees was markedly depressed (0.9 +/- 0.5 mmHg/ml) and was not correlated with established systolic or diastolic indices except for peak positive dP/dt. Our results showed that abnormalities of relaxation were not correlated with contractile dysfunction in patients with DCM in the resting steady state; however, most patients (79%) had both prolonged relaxation and extremely depressed contractility. Abnormal relaxation may have been transient or compensated in some patients (21%) with DCM who had preserved relaxation despite severely depressed contractility. PMID- 8723603 TI - Nonlipid primary and secondary prevention strategies for coronary heart disease. AB - Widespread application of proven primary and secondary preventive strategies for coronary heart disease would result in substantial savings of life and health care dollars. Proven strategies (excluding lipid therapy) include quitting smoking, treating hypertension, physical activity, aspirin therapy, and appropriate use of anticoagulants, beta blockers, and angiotensin-converting enzyme inhibitors in survivors of myocardial infarction. Estrogen replacement therapy is currently under clinical investigation. Avoidance of obesity and tight control of diabetes are prudent interventions as yet unproved by clinical trials. Unfortunately, preventive strategies are frequently underutilized. The greatest challenge for preventive cardiology is to put into practice what we already know to prevent the development and progression of atherosclerosis. PMID- 8723604 TI - Chronotropic incompetence--Part I: Normal regulation of the heart rate. AB - Knowledge of normal adjustment of heart rate and its response to exercise is essential for understanding and management of chronotropic incompetence. The autonomic nervous system plays an important role in the modulation of normal heart rate. Chronotropic responses of a normal heart to exercise are associated with parallel hemodynamic changes in order to meet the metabolic demand of the body. Determination of chronotropic incompetence is widely based on the assessment of maximal heart rate. However, maximal effort should always be confirmed before an attempt to measure a maximal heart rate is made. PMID- 8723605 TI - An unusual case of cyanotic heart disease in a patient with patent foramen ovale and right ventricular hypertrophy. AB - An unusual case of cyanosis and patent foramen ovale/atrial septal defect with right-sided septal hypertrophy is presented. Mild right ventricular dysfunction led to predominant right-to-left shunting. The atrial defect was surgically closed. The patient is from a family with hypertrophic cardiomyopathy; she is negative for the gene abnormality. PMID- 8723606 TI - Hemopericardium with cardiac tamponade after intravenous thrombolysis for acute myocardial infarction. AB - This report presents 2 patients with hemopericardium and tamponade encountered among a group of 26 patients who had received intravenous thrombolytic therapy for acute myocardial infarction during an 8-month time span. Both responded promptly to pericardiocentesis and pericardial catheter drainage. It is suggested that this rarely reported complication has the potential of occurring more frequently with increasing use of thrombolytics followed by aggressive anticoagulant regimens. PMID- 8723607 TI - Monitoring right-sided chest leads during chest pain in patients with obstructive lung disease. PMID- 8723608 TI - Intramural hematoma of the aorta caused by a penetrating atheromatous ulcer. AB - Aortic dissection is a life-threatening condition which is difficult to diagnose because of its variable clinical presentation and diverse pathophysiologic manifestations. We present a case in which intramural hematoma of the thoracic aorta occurred in the absence of an intimal flap. Computed tomography and magnetic resonance imaging both showed hemorrhage in the wall of the aorta. Transesophageal echocardiography also revealed a penetrating atherosclerotic ulcer in the transverse aorta as a cause of the hematoma. A high index of clinical suspicion and a combination of diagnostic procedures resulted in proper diagnosis and successful surgical treatment. PMID- 8723609 TI - Niels Stensen. PMID- 8723610 TI - Building national electronic medical record systems via the World Wide Web. AB - Electronic medical record systems (EMRSs) currently do not lend themselves easily to cross-institutional clinical care and research. Unique system designs coupled with a lack of standards have led to this difficulty. The authors have designed a preliminary EMRS architecture (W3-EMRS) that exploits the multiplatform, multiprotocol, client-server technology of the World Wide Web. The architecture abstracts the clinical information model and the visual presentation away from the underlying EMRS. As a result, computation upon data elements of the EMRS and their presentation are no longer tied to the underlying EMRS structures. The architecture is intended to enable implementation of programs that provide uniform access to multiple, heterogeneous legacy EMRSs. The authors have implemented an initial prototype of W3-EMRS that accesses the database of the Boston Children's Hospital Clinician's Workstation. PMID- 8723611 TI - Relationships among performance scores of four diagnostic decision support systems. AB - OBJECTIVE: To examine the relationships among different performance scores for each of four diagnostic decision support systems (DDSSs). DESIGN: Intercorrelations among seven performance scores on a set of 105 cases for each of four DDSSs (DXplain, Iliad, Meditel, QMR) were computed. METHODS: The performance scores for each case reflected: 1) presence or absence of the case diagnosis in the DDSS knowledge base; 2) presence or absence of the correct diagnosis anywhere on the DDSS diagnosis list; 3) presence or absence of the correct diagnosis in the top ten diagnoses; 4) relevance of the DDSS diagnosis list; 5) comprehensiveness of the DDSS diagnosis list; 6) whether the DDSS suggested additional diagnoses to the experts' list; and 7) the length of the DDSS diagnosis list. RESULTS: For all DDSSs, the two Correct Diagnosis scores (top ten and total list) were significantly related: 1) to the presence of the correct diagnosis in the knowledge base; 2) to the Comprehensiveness score; and 3) to each other. There were significant differences among the four DDSSs on the magnitude and/or direction of the relationships between: 1) the two Correct Diagnosis scores; 2) the Relevance and Length scores; and 3) the Relevance and Additional Diagnoses scores. CONCLUSION: The production of a correct diagnosis for a given case is not related to the number of diagnoses suggested by the DDSS and, across different DDSSs, is not consistently related to other measures of performance. These data indicate that multiple measures are needed to fully describe the performance of a DDSS. PMID- 8723612 TI - Monitoring expert system performance using continuous user feedback. AB - OBJECTIVE: To evaluate the applicability of metrics collected during routine use to monitor the performance of a deployed expert system. METHODS: Two extensive formal evaluations of the GermWatcher (Washington University School of Medicine) expert system were performed approximately six months apart. Deficiencies noted during the first evaluation were corrected via a series of interim changes to the expert system rules, even though the expert system was in routine use. As part of their daily work routine, infection control nurses reviewed expert system output and changed the output results with which they disagreed. The rate of nurse disagreement with expert system output was used as an indirect or surrogate metric of expert system performance between formal evaluations. The results of the second evaluation were used to validate the disagreement rate as an indirect performance measure. Based on continued monitoring of user feedback, expert system changes incorporated after the second formal evaluation have resulted in additional improvements in performance. RESULTS: The rate of nurse disagreement with GermWatcher output decreased consistently after each change to the program. The second formal evaluation confirmed a marked improvement in the program's performance, justifying the use of the nurses' disagreement rate as an indirect performance metric. CONCLUSIONS: Metrics collected during the routine use of the GermWatcher expert system can be used to monitor the performance of the expert system. The impact of improvements to the program can be followed using continuous user feedback without requiring extensive formal evaluations after each modification. When possible, the design of an expert system should incorporate measures of system performance that can be collected and monitored during the routine use of the system. PMID- 8723613 TI - The content coverage of clinical classifications. For The Computer-Based Patient Record Institute's Work Group on Codes & Structures. AB - BACKGROUND AND OBJECTIVE: Patient conditions and events are the core of patient record content. Computer-based records will require standard vocabularies to represent these data consistently, thereby facilitating clinical decision support, research, and efficient care delivery. To address whether existing major coding systems can serve this function, the authors evaluated major clinical classifications for their content coverage. METHODS: Clinical text from four medical centers was sampled from inpatient and outpatient settings. The resultant corpus of 14,247 words was parsed into 3,061 distinct concepts. These concepts were grouped into Diagnoses, Modifiers, Findings, Treatments and Procedures, and Other. Each concept was coded into ICD-9-CM, ICD-10, CPT, SNOMED III, Read V2, UMLS 1.3, and NANDA; a secondary reviewer ensured consistency. While coding, the information was scored: 0 = no match, 1 = fair match, 2 = complete match. RESULTS: ICD-9-CM had an overall mean score of 0.77 out of 2; its highest subscore was 1.61 for Diagnoses. ICD-10 scored 1.60 for Diagnoses, and 0.62 overall. The overall score of ICD-9-CM augmented by CPT was not materially improved at 0.82. The SNOMED International system demonstrated the highest score in every category, including Diagnoses (1.90), and had an overall score of 1.74. CONCLUSION: No classification captured all concepts, although SNOMED did notably the most complete job. The systems in major use in the United States, ICD-9-CM and CPT, fail to capture substantial clinical content. ICD-10 does not perform better than ICD-9-CM. The major clinical classifications in use today incompletely cover the clinical content of patient records; thus analytic conclusions that depend on these systems may be suspect. PMID- 8723614 TI - The accuracy of medication data in an outpatient electronic medical record. AB - OBJECTIVE: To measure the accuracy of medication records stored in the electronic medical record (EMR) of an outpatient geriatric center. The authors analyzed accuracy from the perspective of a clinician using the data and the perspective of a computer-based medical decision-support system (MDSS). DESIGN: Prospective cohort study. METHODS: The EMR at the geriatric center captures medication data both directly from clinicians and indirectly using encounter forms and data-entry clerks. During a scheduled office visit for medical care, the treating clinician determined whether the medication records for the patient were an accurate representation of the medications that the patient was actually taking. Using the available sources of information (the patient, the patient's vials, any caregivers, and the medical chart), the clinician determined whether the recorded data were correct, whether any data were missing, and the type and cause for each discrepancy found. RESULTS: At the geriatric center, 83% of medication records represented correctly the compound. dose, and schedule of a current medication; 91% represented correctly the compound. 0.37 current medications were missing per patient. The principal cause of errors was the patient (36.1% of errors), who misreported a medication at a previous visit or changed (stopped, started, or dose-adjusted) a medication between visits. The second most frequent cause of errors was failure to capture changes to medications made by outside clinicians, accounting for 25.9% of errors. Transcription errors were a relatively ucommon cause (8.2% of errors). When the accuracy of records from the center was analyzed from the perspective of a MDSS, 90% were correct for compound identity and 1.38 medications were missing or uncoded per patient. The cause of the additional errors of omission was a free-text "comments" field-which it is assumed would be unreadable by current MDSS applications-that was used by clinicians in 18% of records to record the identity of the medication. CONCLUSIONS: Medication records in an outpatient EMR may have significant levels of data error. Based on an analysis of correctable causes of error, the authors conclude that the most effective extension to the EMR studied would be to expand its scope to include all clinicians who can potentially change medications. Even with EMR extensions, however, ineradicable error due to patients and data entry will remain. Several implications of ineradicable error for MDSSs are discussed. The provision of a free-text "comments" field increased the accuracy of medication lists for clinician users at the expense of accuracy for a MDSS. PMID- 8723615 TI - Focus on the frontiers of informatics: call for papers on Telehealth and the Informatics of Medical Imaging. PMID- 8723616 TI - ATSDR evaluation of health effects of chemicals. II. Mirex and chlordecone: health effects, toxicokinetics, human exposure, and environmental fate. AB - This document provides public health officials, physicians, toxicologists, and other interested individuals and groups with an overall perspective of the toxicology of mirex and chlordecone. It contains descriptions and evaluations of toxicological studies and epidemiological investigations and provides conclusions, where possible, on the relevance of toxicity and toxicokinetic data to public health. Additional substances will be profiled in a series of manuscripts to follow. PMID- 8723617 TI - [The general principles of cellular gravisensing]. AB - A review of the results of studies over many years into cellular mechanisms underlying gravireception in different cell types, which were carried out by the author in the laboratory during simulation of the effects of altered gravity using a clinostat and centrifuge and under the conditions of space flight. Comparative analyses of the obtained results with the published data has been performed. It was shown that besides specific features of adaptation of different cell types to altered gravity, including microgravity, there are common pathways of adaptation of the structural-functional organization of the cell to this factor. An integral molecular mechanism underlying cell gravisensing was revealed. A working hypothesis on general principles of cell gravisensing has been formulated. PMID- 8723618 TI - [The dose-dependent effects of a combination of different classes of antioxidants exemplified by dibunol and beta-carotene]. AB - In order to enhance antioxidant protection of the organism in various pathological states, effective combinations of antioxidants have been developed that interact with various types of free radicals. On two experimental models,- acute alcohol intoxication and calcium chloride arrhythmia of rats, the prophylactic activity of antioxidant combinations was established for certain doses, rather then for monotherapy. The possibility of obtaining high protective effects at relatively low concentrations of the components in the complex preparation was shown. Am increase of antioxidant concentration in the preparation could decrease its efficiency. The highest efficiency in the complex preparations, as compared with in each component taken separately, was observed in the case of calcium chloride arrhythmia when dibunol and beta-carotene were combined at concentrations of 10 mg/kg and in the case of acute alcohol intoxication (1 and 0.25 mg/kg, respectively). Enhanced efficiency of combined antioxidant therapy opens up the way to the production of new active complex preparations with minimum toxic side toxic effects and complications. PMID- 8723619 TI - [The role of sensory systems in the behavior of Ryukyu mice (Mus caroli Banhote, 1902)]. AB - The role of vibrissa sense of touch, sense of smell, and vision in the Ryukyu mice in experiments with the optic boundary and the glass corridor were studied. A total of 20 adult and 20 young mice were used in the experiments. At the first stage, observations on the intact animals were performed; the animals with removed vibrissa and anosmic animals were used at the second and third stages, respectively. Significant similarities were observed between the behavior of adult males and females. More essential differences were found between the young and adult mice. The role of vibrissa sense of touch is important for orientation of Mus caroli, especially in young mice. Adult animals with a great life experience compensate for the absence of vibrissae more successfully through the intensified use of vision. Anosmia markedly reduces the motor activity of animals and leads to a reduced marking behavior. However, it stimulates the intense use of vision to a lesser extent than does vibrissectomy. PMID- 8723620 TI - [The development of the organum vomeronasale system in human embryos 19--40 mm in crown-rump length]. AB - Age-related changes in the organon vomeronasalis (VO) and related structures have been described in normal human embryos of 19, 25, 28 and 40 mm in crown-rump length and in the embryo of 19 mm in crown-rump length during abnormal development in the uterine tube. The fourth branch of the VO nerve appeared in the 25 mm embryo. Signs of asymmetry and degenerative VO changes were analyzed in the 25 and 40 mm embryos. A supposed vomeronasal ganglion has been described. The possibility of fetuses at the late stages of prenatal development using the VO system is discussed. PMID- 8723621 TI - [Sexual differences in the social behavior of lower catarrhine monkeys (exemplified by grooming in stumptail macaques)]. AB - The structure of social relationships was studied in three groups of stumptail macaques: two groups consisted of macaques of different sexes and one group consisted of males only. In the all-male group, as compared to the groups with different sexes, the social hierarchy was less strict and the social status and resulting grooming depended to a great extent on the age. The attractiveness of individuals as objects was the most variable aspect of grooming. The attractiveness increased with age, while individual preferences were less distinct. PMID- 8723622 TI - [The craniometric characteristics of the elk (Alces alces L.) of Eurasia]. AB - Comparative craniometric investigation was done on the elk in Eurasia. Four morphological forms are distinguished, thus favoring the subdivision of Alces alces L. into four subspecies: A. a. alces, A. a. cameloides, A. a. pfizenmayeri, and A. a. buturlini. PMID- 8723623 TI - [Lipids on the hair surface and the waterproof properties of the mammalian fur coat]. AB - Lipid-like substances incorporated in keratinized parts of the hair or appearing on its surface as sebaceous secretion act as detergents in water. At a sufficient concentration they decrease water surface tension to 0.03 N/m, and, contrary to widespread opinion, should contribute to fur wetting. The influence of these lipids on water surface tension markedly varies among both semiaquatic and land mammals, but in any case their natural concentration is too low to significantly affect the waterproof properties of the cover of the swimming animals. PMID- 8723624 TI - [Comparative genetics of representatives of the genus Yersinia and evolution of their pathogenicity]. AB - This paper was prepared on the basis of a report presented at the first Congress of the All-Russian Society of Geneticists and Breeders (December 1994, Saratov, Russia). Analysis of published data and the authors' own investigations enabled them to propose a concept of evolution of pathogenicity within the Yersinia genus. They concluded that phenotypic features conferred by yersinia plasmids should be considered taxonomic traits. PMID- 8723625 TI - [Cosmid libraries containing DNA from human chromosome 13]. AB - We characterized two cosmid libraries constructed from flow-sorted chromosome 13 at the Imperial Cancer Research Fund (ICRF), UK (13,000 clones) and Los Alamos National Laboratory (LANL), USA (17,000 clones). After storage for two years, clones showed high viability (95%) and structural stability. EcoR I and Hind III restriction patterns were studied in more than 500 ICRF and 200 LANL cosmids. The average size of inserts was shown to be 35-37 kb in both the libraries. Most cosmids (83% and 93% of ICRF and LANL libraries, respectively) exceed the lower size limit of DNA fragments that can be packaged and represent a good source for physical mapping of chromosome 13. Total length of inserts is four and five genome equivalents in the ICRF and LANL libraries, respectively. ICRF cosmids showed hybridization to 22 of 24 unique probes tested, which corresponds to a 90% probability of having any DNA fragment represented in the library. More than 1 Mb of chromosome 13 is overlapped by 90 cosmids of 22 groups revealed. A chromosomal region of more than 150 kb, containing the ATP1AL1 gene for alpha-1 peptide of Na+, K(+)-ATPase, is covered by 12 cosmids forming a contig. The results of restriction and hybridization analyses are stored in a CLONE database. These data and all the cosmids described are publicly available. PMID- 8723626 TI - [Effect of specific nucleotide sequences on the transcription activity of long terminal repeats of avian retroviruses]. AB - Influence of specific nucleotide sequences on the constitutive transcriptional activity of the Rous sarcoma long terminal repeats (LTR) was studied. The possible function of different elements of the transcriptional control region of the avian retroviral LTR was discussed on the basis of a comparative analysis of the effect of specific nucleotide insertions on LTR activity in vivo and in vitro. PMID- 8723627 TI - [Selection and properties of totally phage-resistant mutant Pseudomonas putida PpG1]. AB - The efficiency of using bacteria in open systems to degrade different anthropogenic toxic pollutants can depend strongly on the interaction between these bacteria and natural bacteriophages. The possibility of selecting bacterial Pseudomonas putida mutants resistant to all bacteriophages of this species known so far was tested (in our work, these mutants were designated totally phage resistant mutants). In a model experiment, changes in the composition of a population upon prolonged growth of bacteria in the presence of one of the virulent phages were examined. On the basis of the results obtained, it is postulated that: (1) Mutants differing in resistance to various phages accumulate in a population; relative numbers of different mutants can undergo alterations over the course of time; mutants selected in the presence of a given virulent phage do not often manifest complete resistance to this phage. (2) It is possible to isolate totally phage-resistant mutants of P. putida PpG1. These mutants carry up to three different mutations simultaneously; however, these mutants regain sensitivity to many phages upon pseudoreversion occurrence. PMID- 8723628 TI - [Bioluminescent analysis of the SOS-response of Escherichia coli cells]. AB - We constructed a recombinant plasmid pPLS-1 to estimate the level of SOS response in Escherichia coli by the bioluminescent method. A 6.7-kb promoterless operon of bioluminescence from Photobacterium leiognathi was cloned into a pBR322 vector, in which its expression was controlled by the SOS promoter of gene cda from a plasmid ColD. The sequence between the 5'-terminal Sph1 site of the operon and start codon ATG of the luxC gene was shown to be 56 bp in length and had no effect on the level of light emission. SOS-inducing potency of six mutagenic substances was tested in E. coli strain C600(pPLS-1). The bioluminescent method proved to be very sensitive for estimating the level of SOS response. The results obtained by this method showed good correlation with results obtained by SOS Chromotest, umu-test, and Ames' test. PMID- 8723629 TI - [A strain of the yeast Saccharomyces cerevisiae for testing environmental mutagens based on interaction of the mutations rad2 and him1]. AB - The interaction between mutations at the RAD2 and HIM1 genes was studied. The RAD2 gene encodes endonuclease involved in nucleotide excision repair. Mutants at this gene are highly sensitive to the lethal effect of a variety of mutagens. The product of the HIM1 gene is needed for correction of mismatched bases and repair of premutational DNA damage. Mutations in this gene lead to the formation of the mutator phenotype and high sensitivity to induced mutagenesis. The double rad2 him1 mutant manifested the synergic type of interaction. The level of UV-induced mutagenesis in the double mutant was five times higher than in single mutants, and the absolute yield of forward mutations in five genes controlling adenine biosynthesis was 1 to 2%. UV-induced mutagenesis was increased, at low doses, by several orders of magnitude in the double mutant, compared to the wild-type strain. The high level of mutagenesis in this mutant was caused by ethyl and methyl methanesulfonate. These properties of the stock with the double rad2 him1 mutation makes it promising as a tester in analysis of the gene toxicity of different substances. PMID- 8723630 TI - [Polymorphism of biochemical markers in European bison (bison bonasus)]. AB - The data on 16 polymorphic systems (22 loci) of blood biochemical markers were studied in different breeding lines of European bison: Lowland, Lowland Caucasian, and Mountain lines. Polymorphism was revealed in three loci: Pgm, Idh, and Tf. The average heterozygosity was 3.95%. Statistically significant deviation from the Hardy-Weinberg equilibrium towards an excess of heterozygotes was revealed in the Pgm-2 locus. This suggests certain processes that prevent the decrease of population genetical diversity. The estimations obtained of the genetic distances between European bison lines demonstrated the close relationship among them. PMID- 8723631 TI - [Selection in vivo in cell populations of rat RA-23 rhabdomyosarcoma at the trait "frequency of cells with bridges"]. AB - Selection in vivo of cell clones of rat transplantable organospecific rhabdomyosarcoma RA-23 for increased and decreased frequencies of cells with chromosomal bridges (FCB) was performed. The initial average FCB in clones was 0.8% at a range of variation of from 0 to 3.0%. Selection for an increase, as well as for a decrease, in the FCB in RA-23 clones was effective. After one step of selection for an increase in the FCB, the average FCB increased to 3.0%. Thereafter, selection for an increase in the FCB was impracticable due to loss of transplantability in cell populations in which the FCB exceeded 5.0%. Over five steps of selection for a decrease in the FCB, the average FCB significantly decreased to 0.3% (P < 0.001). The heritability coefficient h2 of the trait FCB upon selection ranged from 0.25 to 0.30. The high and low FCBs attained by selection correlated with the index "frequency of cells with micronuclei." The population of RA-23 cells after selection for a decrease in the FCB differed from the population of cells of the original RA-23 strain by significantly lower karyotypic heterogeneity. The values of h2 obtained upon selection for the FCB and the effectiveness of selection for an increase, as well as for a decrease, in the FCB show that, with selection at the trait "frequency of cells with bridges," which characterizes the stability of the karyotype, existing cell strains can be subjected to karyotypic stabilization or destabilization. PMID- 8723632 TI - [Cytogenetic effect of volatile components of urine of sexually mature animals on bone marrow cells of young female house mice Mus musculus L]. AB - Effects of volatile components of the urine (VCU) of mature male and female CBA/LacStoRap mice on proliferating bone marrow cells in young females of the same line were analyzed. To estimate the effects of VCU, cytogenetic analysis of mitotic disturbances in anaphase-telophase cells was performed. The overall frequency of mitotic disturbances such as chromosome bridges, fragments, retarded chromosomes, and multiple rearrangements was assessed. The overall frequency of mitotic disturbances in young female mice treated with VCU from mature females for 2 h significantly increased 24 h after exposure. The spectrum of mitotic disturbances was also affected by the VCU treatment. After exposure to VCU from mature males, no significant variation of parameters analyzed was detected in young female mice. The possible mechanisms and role of specific effects of VCU from animals of different sexes on young female house mice are discussed. PMID- 8723633 TI - [Genetic study of idiopathic torsion dystonia in Russia]. AB - Inheritance of idiopathic torsion dystonia (ITD) was studied in 41 Russian families including 41 probands with generalized, focal, and segmental dystonia and 140 recurred cases. Affected relatives appeared in two or more generations in 31 families analyzed. It was shown that in 76% of segregated cases, ITD was inherited as an autosomal dominant trait with a penetrance of 40% and varying expression. An autosomal recessive type was observed in 24% of the cases. Approximately 10% of the cases of disease could be caused by a new mutation and 14.6% by a nongenetic phenotype similar to genetic forms in its clinical symptoms. ITD with the X-linked recessive type of inheritance did not occur in the families studied. The recurrence risk was 20% in autosomal dominant forms. The risk correlated with age the relative's: clinical symptoms developed in 98.4% of patients by the age of 30. PMID- 8723634 TI - [Polymorphism of placental alkaline phosphatase on the level of DNA and protein in the Mordovian population]. AB - Data on DNA and enzyme polymorphisms of human placental alkaline phosphatase (PLAP) in Mordvinian populations are presented. Restriction fragment length polymorphism (RFLP) was detected after the digestion of DNA samples with Rsa I and Pst I endounucleases. The frequencies of the second (2) allele for PLAP Pst I and Rsa I were 0.53 and 0.192, respectively. Comparative data suggest that there are no population differences between Mordvinians and Scandinavian ethnic groups. In Mordvinians gene frequencies measured at the level of gene products were PLAP*1(S) = 0.681, PLAP*2(F) = 0.244, PLAP*3(I) = 0.069, and PLAP*18(D) = 0.006, indicating the similarity of the corresponding values in Scandinavians. The observed RFLP and "protein" genotype frequencies were in good agreement with that expected according to the Hardy-Weinberg equation. In the Mordvinian population, as in those, surveyed previously, a strong linkage disequilibrium between Pst I and Rsa I PLAP alleles was observed. PMID- 8723635 TI - [Population genetics of spinal muscular atrophy]. AB - A population genetic study of spinal amyotrophy (SMA) in six Russian and three Central Asian regions was carried out. In total, 29 patients with autosomal recessive (AR) infantile proximal SMA (SMA I-III) and four patients with rare SMA forms with an unspecified type of inheritance were revealed. In Russian populations, the prevalence of SMA I-III is similar (1.5-2.5/100000), it is one of the most common hereditary neurological diseases. A tendency toward nonuniform territorial SMA prevalence is observed in genetically subdivided populations. The lesser SMA I-III prevalence in Central Asian populations might be due in part to inbreeding depression. A segregation frequency of 0.21 is in accordance with AR inheritance; the proportion of sporadic cases is 3%. Clinical genealogical data support the genetic unity of forms I-III. The origin of pedigrees with SMA in distant relatives is discussed. PMID- 8723636 TI - [Mitochondrial DNA variation in native inhabitants of Siberia with reconstructions of the evolutional history of the American Indians. Restriction polymorphism]. AB - Restriction polymorphism of mitochondrial DNA (mtDNA) isolated from the blood of 107 Koryaks, 28 Northern Altaians, and 23 Kets was studied. The results were compared with similar data that in aggregate characterized haplotypes of 569 individual DNAs from 13 Siberian populations, in order to elucidate the genetic similarity and differences between mtDNA haplogroups of native Siberians and Americans. Natives of Siberia are distinguished by the significant variation of the composition and frequency of three (A, C, and D) of four haplogroups characteristic of American Indians. In the full composition, all four haplogroups A, B, C, and D-are found only in a sample of Northern Altaians. The peculiarities of their geographical distribution in Asia allow us to consider the southern regions of Siberia and the adjacent territories of Central and East Asia as the site inhabited by the ancestors of the first wave of migrants to North America. PMID- 8723637 TI - [Marital structure of the Kursk population]. AB - Age characteristics, national composition, migration processes, and the level of inbreeding among Kursk citizens were analyzed on the basis of a sampling study of marriages registered in Kursk in 1960 and 1989. Assortative marriages in reference to age were r = 0.910 in 1960 and r = 0.904 in 1989. The polychoric relationship index characterizing national correlation of married pairs was K = 0.440 in 1960 and K = 0.405 in 1989. Positive assortive mating in reference to the birthplace of the married pairs was revealed: K = 0.190 in 1960; K = 0.161 in 1989. The coefficient of local population relationship (a) was 0.000026 in 1960 and 0.000011 in 1989. PMID- 8723638 TI - [Differential characteristics of the temperature-sensitive period of the lawcp1 mutation on the basis of macrochaetae overexpression in Drosophila]. AB - Temperature-sensitive periods (TSPs) of lawcpl mutations were studied via the frequencies of arising of extras macrochaetae for each macrochaetae type separately. TSPs for the formation of extras macrochaetae of original classes were unevenly distributed within larval instars II and III. It was found that macrochaetae of different types differed from one another in their temperature sensitivity. The TSP of development of macrochaetae belonging to new ectopic classes was multiphase, with the first phase at larval instar II, and the second phase, at the prepupal to early pupal stage. It is assumed that in Drosophila, the products of the lawc gene participate, along with other genes, in the development of the nervous system through regulation of ac-sc expression, both in proneural clusters and outside them. PMID- 8723639 TI - [Fingerprinting of potato species and cultivar using RAPD-PCR]. AB - Polymerase chain reaction with random primers (RAPD-PCR) is used to fingerprint DNA polymorphism of six species and eight cultivars of potato. Three primers were developed experimentally for detection by DNA fingerprinting of cultivars and species. High significance and reproducibility of amplified DNA spectra for each species and cultivar was demonstrated. In these spectra, RAPD-PCR fragments, were revealed which may serve as cultivar-and species-specific markers for identification and differentiation of Solanum cultivars and species. PMID- 8723640 TI - [Effect of a single-locus mutation of the silver-blue color allele in mink on the dopaminergic system of the brain]. AB - The peculiarities of dopamine metabolism and dopamine receptors in the brain of homozygous silver-blue (pp) minks were examined in comparison with wild-type (PP) minks. A pp mutation was found to affect the dopamine turnover in corpus striatum, which is one of the major dopaminergic brain structures. In silver-blue minks, increased dopamine catabolic enzyme activity monoamine oxidase type B (MAO B), together with the decreased level of dopamine and the tendency for the level of the metabolite 3,4-dihydrophenyl acetic acid to increase, were demonstrated in this brain region. No significant alterations in the level of another dopamine metabolite, homovanillic acid, or in D1 and D2 receptor density were shown. In the midbrain and hippocampus, an elevated level of MAO B activity was detected. We found no modifications in receptor properties, nor in dopamine turnover in a mesolimbic dopamine structure, the nucleus accumbens. We propose that the primary effect of pp mutation is an increase in MAO B activity resulting in dopamine turnover alterations and probably in dopamine-dependent behavior (pleiotropic cascade). PMID- 8723641 TI - [Increase in the frequency and risk of birth of children with Down syndrome in Leningrad (1982-1989)]. PMID- 8723642 TI - Roles of dopamine receptors in long-term depression: enhancement via D1 receptors and inhibition via D2 receptors. AB - The effects of both the activation and the blockade of D1 or D2 dopamine receptors on long-term depression (LTD) of synaptic transmission, and the involvement of NMDA and GABA receptors in LTD, were investigated in CA1 neurons of rat hippocampal slices. Low-frequency stimulation (LFS, 450 pulses at 1 Hz) produced LTD of the slope of field EPSPs (-14.3%, mean, n = 10). The induction of LTD was blocked by the NMDA receptor antagonist, AP5 (1.4%, n = 7), by the D1 receptor antagonist, SCH-23390 (3.5%, n = 8), or by the D2 receptor agonist, LY 171555 (4.4%, n = 8). Either the activation of D1 receptors by SKF-38393 or the blockade of D2 receptors by sulpiride produced significantly larger LTD than the control LTD (-31.1%, n = 11; -30.6%, n = 9, respectively). Although LTD was blocked by picrotoxin, a GABAA receptor/Cl- channel antagonist (4.9%, n = 8), LTD was produced by LFS in the medium containing both SKF-38393 and picrotoxin ( 27.3%, n = 7). These results indicate that: (1) the induction of LTD by LFS in hippocampal CA1 neurons is under the influence of both NMDA and GABA receptors; (2) both D1 and D2 receptors are involved in the modulation of LTD in that the activation of D1 receptors enhances LTD, while that of D2 receptors inhibits LTD, and (3) while the induction of LTD is blocked by picrotoxin, this effect is superseded by SKF-38393. PMID- 8723644 TI - The alpha and omega of G-protein coupled receptors: a novel method for classification. Part 1. AB - In this paper, a novel method is presented for classification of 113 neurotransmitter and opioid receptors that is based on the counting of amino acid residues. With the use of sequence alignment, ten conserved key residues were identified: alpha (the first Met residue of the sequence), Asn in the tentative first transmembrane domain (TM1), Asp (TM2), Cys (top of TM3), Arg (bottom of TM3), Trp (TM4), Cys (in the loop between TM4 and TM5), Pro (TM5), Pro (TM6), Pro (TM7) and omega (the last residue of the sequence). The number of residues between these key residues is unique for each receptor or receptor subtype and is used for classification. The number of residues between two key residues defines a segment. The sum of the segments before or after a key residue is defined as a partition. In total, 73 possible classification schemes were found using two or three segments, partitions or a combination of segments and partitions. The surprising and striking results is that each of the sequences examined can be characterized by a code consisting of two or three figures. Each figure represents a number of amino acid residues. PMID- 8723643 TI - The amino terminal domain of the glucagon-like peptide-1 receptor is a critical determinant of subtype specificity. AB - Glucagon-like peptide-1 (GLP-1) and glucagon are peptide hormones involved in glucose homeostasis. The ligands are closely related (48% identical) and bind with different affinities to distinct, although highly homologous (47% identical), G protein coupled receptors on the surface of cells. By these criteria, glucagon and GLP-1 receptors can be considered receptor subtypes. A series of chimeric receptors in which 4-6 amino acids in the N-terminal extracellular domain of the human GLP-1 receptor were replaced with the analogous region of the human glucagon receptor were constructed and expressed in COS-7 cells. One of these chimeric receptors, C29-32 displays a 7 to 10-fold decrease in affinity for GLP-1 and the GLP-1 antagonist exendin 9-39 amide and a concomitant 7 to 9-fold increase in its affinity for glucagon. This change in affinity results in a 50-fold decrease in the selectivity of this receptor for GLP-1 over glucagon. Thus, the substitution of as few as four residues of the GLP 1 receptor profoundly affects its selectivity for the homologous peptide agonists GLP-1 and glucagon. These results suggest the extracellular N terminal domain of the GLP-1 receptor harbours molecular determinants for both agonist binding affinity and selectivity. PMID- 8723645 TI - A unitary non-NMDA receptor short subunit from Xenopus: DNA cloning and expression. AB - A high-affinity homomeric, non-NMDA glutamate receptor was previously purified from the amphibian Xenopus laevis. We have obtained nine peptide sequences from its subunit, applied in cDNA cloning. The cDNA encodes a subunit (XenU1) containing all nine sequences. The 51,600-dalton mature subunit has four hydrophobic domains homologous to the four in the C-terminal half of mammalian non-NMDA receptor subunits. Transient expression in COS cells showed 1:1 binding (at Bmax) of [3H] kainate (KD = 9.1 nM) and of [3H] AMPA (alpha-amino-3-hydroxy-5 methyl-isoxazole-4-propionic acid; KD = 62 nM). The competitive binding series domoate > kainate > AMPA > NBQX > glutamate was established (where NBQX is 2,3 dihydroxy-6-nitro-7-sulphamoyl-benzo (f) quinoxaline). Each agonist shows the same KI value against [3H] kainate and [3H] AMPA binding, suggesting a common agonist site, but two conformations thereof are distinguishable by their different affinities for the antagonist NBQX and by the allosteric effect of thiocyanate anion (greatly potentiating AMPA binding, inert with kainate). XenU1 is exceptional among non-NMDA receptor subunits because it lacks most of the large N-terminal domain found in those of mammals and it has high affinity for both kainate and AMPA. It differs from the similarly-short "kainate-binding proteins" (KBPs), in binding AMPA and in forming glutamate receptor channels when the native protein is reconstituted. Moreover, whereas a full-length kainate receptor of mammals, GluR6, is shown here (from a partial cDNA sequence) to exist also in Xenopus, with approximately 97% sequence identity to rat GluR6, XenU1 is much less homologous to any rat kainate or AMPA receptor and also to the KBPs, even from another amphibian, Rana. Another difference is that a potential concensus sequence ("EF hand") for Ca2+ binding is present in the N-terminal domain of XenU1, but not in the chicken (glial) KBP. XenU1 is deduced to be in a new family of non-NMDA receptors. PMID- 8723646 TI - The S. cerevisiae outwardly-rectifying potassium channel (DUK1) identifies a new family of channels with duplicated pore domains. AB - Potassium channel subunits have six or two transmembrane segments in addition to a conserved pore-forming (P) domain; four subunits come together to form a channel. A gene was identified in S. cerevisiae (J0911) encoding a protein with eight probable membrane-spanning segments and two such P regions. This protein (Duk1p) is a potassium channel because Xenopus oocytes injected with the corresponding RNA express potassium currents activated by depolarization that are not seen in control oocytes. Similar potassium currents were recorded from wildtype S. cerevisiae spheroplasts, but not from those in which the DUK1 locus had been disrupted. Cells carrying the duk1 delta 1::HIS disruption in addition to a chimeric gene comprising DUK1 behind the GAL1 promoter showed outward currents when grown in galactose, but not when grown in glucose. Additional sequences with the duplicate pore motif were found in C. elegans, suggesting that these proteins represent a novel structural family of potassium channel proteins. PMID- 8723647 TI - Mutation of R555 in CFTR-delta F508 enhances function and partially corrects defective processing. AB - The most common cystic fibrosis mutation deletes phenylalanine 508 in CFTR (CFTR F508). This mutation causes the loss of CFTR Cl- channel activity by disrupting biosynthetic processing so that mutant protein does not reach the plasma membrane. It also decreases the rate at which mutant channels open. To identify second-site mutations that could reverse the effects of delta F508, we used a chimeric yeast STE6/CFTR system bearing the delta F508 mutation. This chimera manifests defective mating. After mutagenesis of the first nucleotide-binding domain, we found that mutation R555K partially restored mating. However, it also increased mating in the chimera lacking the delta F508 mutation. When we introduced the R555K mutation into human CFTR-delta F508, we found that the loss of apical Cl- current caused by delta F508 was partially restored, predominantly due to a partial correction of the delta F508 processing defect. Analysis of single CFTR Cl- channels showed that the R555K mutation did not correct the prolonged closed time associated with delta F508, rather it increased activity of both wild-type and delta F508 channels by prolonging the duration of bursts of activity. These data suggest that the region around residue R555 in the first nucleotide-binding domain is important both in determining the ability of the delta F508 protein to be properly processed and in determining channel function. PMID- 8723648 TI - [Different character of zinc ion binding with lactate dehydrogenase M4 in diluted and concentrated solutions]. AB - The study of the melting characteristics of lactate dehydrogenase M4 and Zn/lactate dehydrogenase at pH 6.5-8.5 by microcalorimetric methods shows that in dilute solutions of the enzyme (0.05% lactate dehydrogenase solution) one Zn ion causes destabilisation of enzyme structure, attended by a decrease of melting temperature throughout the entire pH interval under study. Concentrated solutions of the enzyme (0.4% lactate dehydrogenase solution) are a seat of such conformational changes of the latter which affect the character of Zn ions binding depending on pH. PMID- 8723649 TI - [Comparative study of the kinetics of phospholipid liposome peroxidation, induced by hypochlorite and in the Fe2+ + ascorbate system]. AB - The present study examined the kinetics of accumulation of lipid peroxidation products in phosphatidylcholine liposomes incubated with hypochlorite (HOCl/OCl-) or with FeSO4 in the presence of ascorbate. The incubation of liposomes with HOCl/OCl- led to the accumulation of primary (lipid hydroperoxides and conjugated dienes) and secondary (TBA-reactive substances) lipid peroxidation products identical to those generated in the presence of Fe2+/ascorbate. However the rate of the formation of lipid peroxidation products as well as the gain in their amount were sufficiently higher (4-150-fold and 3-6-fold, respectively) in the case of HOCl/OCl(-)-induced lipid peroxidation as compared to lipid peroxidation in the Fe2+/ascorbate system. The accumulation of lipid peroxidation products lasted until HOCl/OCl- was present in the reaction mixture. The data obtained support our hypothesis that hypochlorite produced by activated neutrophils and monocyte/macrophages may be reasonable for the initiation of lipid peroxidation in blood. PMID- 8723650 TI - [Kinetics of lipid peroxidation induced by Fe2+ ions in a suspension of mitochondrial and nuclear membranes]. AB - Kinetics of TBARS accumulation, Fe2+ oxidation, and chemiluminescence were measured in nuclear and mitochondrial membrane suspensions during Fe(2+)-induced lipid peroxidation development. Above certain critical Fe2+ concentrations ([Fe2+].) the latent period (t) of lipid peroxidation development were observed. By using the dependence of t on [Fe2+] the [Fe2+]. values were measured in nuclear and mitochondrial membrane suspensions. At the same concentrations of nuclear and mitochondrial membranes (1 mg/ml) the [Fe2+] value changed from 10 to 30 mu kM. In the case of nuclear membrane suspension the [Fe2+] value changed from 20 to 30 mu kM with the change of membrane concentrations from 0.5 to 1.5 mg/ml. For mitochondrial membrane suspension [Fe2+]. value increased from 10 to 40 mu kM with increasing membrane concentrations from 1 to 4 mg/ml. Apparently, [Fe2+]. value is the main parameter determining the kinetics of Fe(2+)-induced lipid peroxidation in native membrane preparations; the [Fe2+]. value in different membrane systems, in turn, is dependent on the ability of the membrane to bind Fe2+ ions. PMID- 8723651 TI - [Interaction of 4-(n-dimethylaminostyryl)-1-methylpyridinium-n-toluolsulfonate with liposomes: analysis of fluorescence spectra]. AB - Using fluorescent probe 4-(n-dimethylaminostyryl)-1-methylpyridinium n toluenesulfonate the radiation effect on the structure of liposomes composed of phosphatidylcholine and diphosphatidylgycerol has been investigated. The from of fluorescence spectra was analyzed and inhomogeneous broadening parameters of spectral components were estimated. The polarity of DSM environment and probe distribution between different sites were found to change after irradiation. PMID- 8723652 TI - [Competitive interaction of the fluorescent probe 4-(n-dimethylaminostyryl)-1 methylpyridinium n-toluolsulfonate and lysozyme with lysosomes]. AB - Competitive binding of fluorescent probe 4-(n-dimethylaminostyryl)-1 methylpyiridinium n-toluenesulfonate and lysozyme to liposomes composed of phosphatidylcholine and diphosphatidylglycerol has been investigated. Association constants and binding stoichiometry for probe and protein have been estimated. Increase of the bilayer negative charge was found to be followed by the increase of binding constant and number of phospholipids interacting with lysozyme. PMID- 8723653 TI - [Changes in binding parameters of fluorescent probes 1-anilinonaphthalene-8 sulfonate and 4-(n-dimethylaminostyryl)-1-methylpyridinium n-toluolsulfonate with liposomes upon radiation exposure]. AB - Using fluorescent probes 1-anilinonaphthalene-8-sulfonate and 4-(n dimethylaminostyryl)-1-methylpyridinium n-toluenesulfonate the effect of radiation on the properties of liposomes composed of phosphatidylcholine and diphosphatidylglycerol has been investigated. Parameters of the probes association with liposomes were estimated. It was suggested that radiation induced increase of 1-anilinonaphthalene-8-sulfonate fluorescence intensity and decrease of that for 4-(n-dimethylaminostyryl)-1-methylpyridinium n toluenesulfonate are caused by lipid's peroxidation. PMID- 8723654 TI - [Role of components of formate-hydrogen-lyase in forming molecular hydrogen and their connection with proton-potassium exchange in anaerobically grown Escherichia coli]. AB - It is shown that -2H+/K(+)-exchange through the H(+)-K(+)-pump, formed by the F0F1-ATPase and the Trk H system, H(+)-K(+)-exchange via H(+)-K(+)-antiporter, formed by the F0 and the Trk G (core) system [1-2], and production of H2 in anaerobically grown E.coli are changed in the mutants with defects in components of formate hydrogen lyase complex, oxidizing formate to CO2 and H2. 2H+/K(+) exchange and H2 production are destroyed, but H(+)-K(+)-exchange with a variable stoichiometry for N,N'-dicyclohexyl-carbodiimide-sensitive ion fluxes is displayed in the fdhF mutant E.coli FM911, where formate dehydrogenase(H) is absent. 2H+/K(+)-exchange does not occur, but H(+)-K(+)-exchange with variable stoichiometry for N,N'-dicyclohexylcarbodiimide-sensitive ion fluxes and H2 production are observed in the uncD mutant E.coli AN817 with defect in beta subunit of the F1. Deletion of the hyc-operon in mutant E.coli HD700, led to absence of hydrogenase 3, destroys H(+)-K(+)-exchange and H2 production. H2 evaluation is shown in the E.coli K12(lambda) protoplasts, treated with toluene, by adding of NADH into the medium, containing ATP and K+. It is inhibited by N,N' dicyclohexylcarbodiimide. H2 production is increased by adding of dithiothreitol, when NADH is changed by formate. It is lost in the mutants with defects in the F0 (E.coli AN936) or in the Trk A protein (E.coli TK2242). Dehydrogenase(H) and hydrogenase 3 are assumed to link mutually with a H(+)-K(+)-pump operation, reducing equivalents, necessary for a dithiol-disulfide interconversion within a mechanism of pump, are transferred from formate by means of dehydrogenase(H) to hydrogenase 3 through the F0F1 and the Trk H system to produce H2. It is assumed that hydrogenase 3 can interact with a mechanism of H(+)-K(+)-antiporter, NADH could serve as a donor of reducing equivalents. A role of thiol-groups and dithiol-disulfide interconversion in a functions of both mechanism for H(+)-K(+) exchange is confirmed. PMID- 8723655 TI - [Character of K+ absorption and its interaction with membrane protein pumps in Escherichia coli, grown under anaerobic conditions in the presence of nitrate]. AB - The character of H(+)-K(+)-exchange in E.coli, grown in anaerobic conditions in the presence of sodium nitrate and performed a nitrate respiration, has been studied. The K+ uptake has been shown to occur by one step, to be not inhibited by N,N'-dicyclohexylcarbodiimide, but to be stopped by arsenate and protonophore. It has K(m) of 4.5 mM, K+ accumulation in cell is more than 200 mM and K+ distribution between the cytoplasm and the medium is more than 10(3) (K+ equilibrium potential is achieved 190 mV). To switch on a mechanism of K+ uptake is depended on osmotic shock and carried out upon positive as well as negative shocks in spheroplasts. A H+ efflux occurs with constant rate while glucose is in the medium and a stoichiometry for the initial H+ to K+ fluxes is variable upon a different experimental conditions. In spite of H(+)-K(+)-exchange in anaerobically grown E.coli, a production of H2 in bacteria is not observed, an ATPase activity of isolated membranes sensitive, to N,N'-dicyclohexylcarbodiimide is not stimulated by K+ and lost in E.coli mutant with an unc-deletion. It is concluded that H+ and K+ transfer through the membranes in E.coli performed a nitrate respiration, occur through different systems-a respiration chain and the TrkA system of K+ uptake. The latest operates itself, has no ATPase activity and interacts with membrane proton pumps indirectly using transmembrane proton gradient (delta mu H+) as a driving force as well as ATP as a regulator of activity. A sensitivity of this system to osmotic shock is lost under destruction of periplasmic space. PMID- 8723656 TI - [Kinetics of nerve ending ion currents with nonhomogeneous distribution of ion channel density]. AB - The Hodgkin-Huxley's equations of unmyelinated nerve fiber's electric behaviour has been used to create model of nerve endings electrogenesis. The Eler's integration method has been applied to solve the partial differential equation system which describes the spread of excitation in nerve ending. The effect of nonhomogeneous distribution densities of ionic channels along nerve ending on the ionic currents were simulated. The comparison of computed curves with nerve ending responses recorded by extracellular electrodes was performed. It was shown that the better match between simulated and recorded responses observes during densities' decrease potential-dependent sodium, potassium and calcium-activated potassium channels according to exponent along the nerve ending without changes of calcium channels density. As the result the amplitude and time-course of spread action potential, calcium current and transmitter release along the nerve ending appear to be changed. PMID- 8723657 TI - [Study of the viscosity of excited membranes using combined dispersion spectroscopy]. AB - Raman spectra and spin-labeled fatty acids of myelinated frog sciatic nerve and mollusc neurons at different temperature was investigated. The correlation between nerve and excitation and microviscosity of the membrane and cytosol lipids was found. This results is interpreted as reflecting an increase of fluidity of the hydrocarbon core of nerve membrane with temperature. Finding may be interpreted as transient "decreased-increased" fluidity of the membrane lipid during rhythmic excitation. PMID- 8723658 TI - [Anomalous dependence of erythrocyte cytosol microviscosity on temperature and composition of the media]. AB - Abnormal decrease in microviscosity in erythrocytes cytosol during temperature decrease has been established when erythrocytes are in hypertonic sodium chloride solution in the presence of 1,2-propane diol and without it and also in the solution of polyethylene glycol-1500. PMID- 8723659 TI - [Effect of erythrocyte membrane structure on the dose dependence of photohemolysis]. AB - The influence of different factors modifying erythrocyte membrane structure on the value of power index of doze dependence of photohemolysis of hematoporphyrin sensitized erythrocytes has been investigated. It has been established that thermoinduced spectrin aggregation results in increase the index, whereas vesiculation under metabolic exhaustion of erythrocytes greatly decreases value n. A more considerable decrease of the parameter was observed when photohemolysis of erythrocytes was occurred in alkaline medium. It has been shown that the observed decrease of the power index in the latter case is not caused by pH induced changes of photochemical reactions but by some dark processes. The decrease is possibly brought about by pH-induced conformational changes of protein band 3. PMID- 8723660 TI - [Autohemolysis of human erythrocytes in the temperature range 4-37 C]. AB - It is shown that the value of activation energy of red blood cells autohemolysis in temperature range of 4 + 37 degrees C is considerably smaller than that at t > 37 degrees C and is about 50 kJ/mol in isotonic saline buffer, pH 7.4. This means that the change of limiting stage of hemolysis occurs at 37 degrees C. It is found that the decreasing of oxygen partial pressure in the medium resulted in marked inhibition of hemolysis without alteration of activation energy in temperature range of 20 divided by 37 degrees C and on the contrary in intensification of hemolysis at t < 20 degrees C. It is concluded that oxidative processes are essential in autohemolysis in temperature range of 20 divided by 37 degrees C and the oxygen stabilizes the structure state of erythrocytes below 20 degrees C. PMID- 8723661 TI - [Mechanisms of the combined effect of SHF electromagnetic radiation and hydrogen peroxide on the viability of microorganisms]. AB - The cellular (viability) and molecular (kinetics of changes in nucleoid viscosity) mechanisms of damage to various strains of microorganisms after single and combined action of SHF electromagnetic radiation and hydrogen peroxide are compared. The conditions are found in which marked synergetic interactions of the agents are realized. The role of repair systems in formation of final products of the interactions is discussed. PMID- 8723662 TI - [Effect of a galvanic current on ischemic damage to the myocardium]. AB - Galvanization of precardial region in rats with experimental infarction elicits considerable cardioprotective effect manifested in a decrease of the mass of necrotic tissue and in partial normalization of ion content both in the necrotic focus and outside it. PMID- 8723663 TI - [Model for isolating spectral maxima of an acoustic signal at the first levels of the hearing system]. AB - The mathematical model of the frequency analysis of acoustic signal, based on the physiological data about that processing in the peripheral part of mammals auditory system is given. The main principle of this model is the use of phase temporal structure of signal in each frequency channel. PMID- 8723664 TI - [Effect of microphone potential feedback on formation of adjusted curves of the hearing periphery]. AB - The model of the formation of the exiting factor on the neuron endings of the cochlear nerve taking into consideration the feedback mechanisms described in recent experimental investigations is proposed. It is shown that the functioning of the feedback in the model give the possibility to explain the principle of action of the mechanism of enhancement of mechanical duct tuning which produce exquisite tuning, reflected in the tuning properties of cochlear nerve fibers. At the same time the utilisation in the model the mechanism of the lateral inhibition provide it the property similar to the effect of inhibition of the neurone response to a given tone signal when another tone signal arises in a nearby frequency region. PMID- 8723665 TI - [Informative value of forms of visual evoked potentials in assessing the location of a visual tract block]. AB - It is proposed the explanation of the form of visual evoke potential (VEP) on the base of physical imagination of the electrical field of double electrical layers that can approximate the wave of excitement of the visual tract. We mast take into consideration the electrical potential as active as indifferent electrode simultaneously and estimate visual evoke potential as diversity. It is shown on mathematical model the more smooth form of visual evoke potential under block and unimportant to arrangement of the electrode over the place of block. PMID- 8723666 TI - [Role of the exterior geniculate bodies and superior colliculus in the system that collects and accumulates visual information]. AB - An analysis of picture on- and off-decompositions by retina ganglion cells and ipsi- and contralateral projections interleaving in lateral geniculate bodies (LGB) layers lead to founding earlier unknown LGB function-the logic filtering of the accurate (common in two approximations) parts of images. On- and off splitting guarantees non-depended picture over and below approximations and reliable performing the above filtering function in the LGB layers. System complementation to LGB function is retinotopic mapping of non-percept pixels. It relates to primary superior colliculus (SC) function. Correlation between LGB and SC functions and saccade eye movements is discussed. Neurological nets for LGB and for a part of SC which realize the specified processor functions are presented. PMID- 8723667 TI - [A model of the respiratory control system]. AB - A modification of the Mackey-Glass model for a respiration control system is proposed. The equations corresponding to the proposed model were built. A solution of the equations was obtained, which corresponds to the Cheyne-Stokes irregular respiration. PMID- 8723668 TI - [Short-period oscillations of isotopic carbon in CO2 from expired air in different human functional states]. AB - The examination of diurnal variations in carbon isotope composition of CO2 under different hormonal-metabolic states in man revealed short-period inphase oscillations with a period of about 2-3 h. The oscillations occur throughout 24 h and their period is little dependent on the functional state of the organism. In endocrine pathologies, substantial differences in oscillation amplitude are observed. The relationship between the variations in carbon isotope composition of CO2 of expired air and energy exchange in cells of insulin-dependent tissues of the organism is substantiated. PMID- 8723669 TI - [Average daily values of the isotopic carbon composition in expired air and urea in human urine in normal states and in some endocrine disorders]. AB - Daily average carbon isotope composition of CO2 of expired air and urine urea in patients being in different hormonal-metabolic states was determined. The observations were carried out under conditions of clinic to provide standard diet, and the same daily and food taking regimes. In all persons, a substantial increase enrichment in 12C of CO2 of expired air (by 3-6%) and 13C of urine urea (by 3-5%) relative to carbon of food was revealed. Marked variations in these characteristics in norm, on starvation and in endocrine pathologies were found. A relationship between the carbon isotope composition and the metabolic shifts characterizing the deviations of the studied functional states from norm was established. The changes in the isotope characteristics and their relation to the hormonal-metabolic status of the organism are interpreted in terms of the model for cell fractionation of carbon isotopes proposed earlier. PMID- 8723670 TI - [Effect of weak SHF radiation on the level of fat soluble vitamins A and E in rat tissues]. AB - The effect of SHF radiation on the content of A and E vitamins in tissues of rat brain, liver and testis was studied. Among the tissues examined, the brain was found to be most sensitive to weak SHF radiation. The radiation induced a change in the ratio of A and E vitamins in the brain, which, in turn, increased the probability of damage to cell membranes. In the liver, the tocopherol level after irradiation remained unchanged, and in the testis it increased insignificantly. This suggests a high resistance of the antioxidant systems of liver and testis tissues. PMID- 8723671 TI - [Probability of a "greenhouse catastrophe" and the nature of feedback]. AB - The chance of "hotbed catastrophe" appearance is discussed on the theoretical base. The feedbacks have essential significance in ecological and evolutional processes. They are directed against the cause, which disturb the status of ecological equilibrium in biosystem always. The "hotbed effect" will provoke biospherical processes, which will be directed to the reconstruction of ecological equilibrium status. "Hotbed catastrophe" chance is very small. PMID- 8723672 TI - [Greenhouse catastrophe and the problem of developing human civilization]. AB - Principal distinctions between the "greenhouse disaster" and traditional scenarios of mean global temperature increase on Earth caused by greenhouse effect are discussed. Processes accompanying "greenhouse disaster" are compared with actual climatic catastrophies of Carbon and Cretaceous. Different factors, such as underground nuclear explosions, are discussed in view of their influence on lithosphere stability and rate of mean global temperature increase. PMID- 8723673 TI - [Greenhouse effect and probable prospects: global catastrophe or optimization of the biosphere?]. AB - Author pay attention to polyvariantness of expectative consequences of hotbed effect, to evolution-biological aspect of problem, and to evolution prognosis difficulties. The perspective of biosphere and mankind survival is mutual adaptation in coevolution process. Creation of complete biological evolution theory is becoming more and more urgent. Evolution prognosis and biosphere management will be realized on evolution theory base. PMID- 8723674 TI - [Greenhouse catastrophe: results of discussion]. PMID- 8723675 TI - [Conformational features of circular DNA with natural curvature]. AB - Monte-Carlo modelling of DNA circles revealed that features of LK distributions must strongly depend on the DNA sequence. The effect is very significant for circles of 300-1500 bp length with a(stt) < or = 250 nm. PMID- 8723676 TI - [Use of polarized thermomicroscopy for recording formation and degrading processes of collagen fibrils]. AB - Polarized thermomicroscopic method were used for registration of collagen fibril formation and thermal degradation processes. It was compared with differential scanning microcalorimetry and optical density measurement methods and recommended as a fast method for registration of collagen fibril formation and degradation processes. PMID- 8723677 TI - Relationship between genotype and phenotype in monogenic diseases: relevance to polygenic diseases. AB - Since the early descriptions of sickle cell anemia, it has been clear that genotype at a single locus rarely completely predicts phenotype. This paper reviews explanations for phenotypic variability in some monogenic diseases. In cystic fibrosis, there is strong correlation between genotype and pancreatic phenotype but only weak association with respiratory phenotype, possibly due to differential inheritance of alleles at loci controlling susceptibility to respiratory infection. In addition, disease mutations have been shown to have more or less severe effect, depending on other variation within the cystic fibrosis gene. In phenylketonuria, genotype at the phenylalanine hydroxylase locus appears to explain the biochemical phenotype, but not the intellectual status. There may be genetically determined variation in flux through the minor metabolic pathways for phenylalanine, influencing levels of alternative metabolites involved in mental development. Phenotypic discordance in sickle cell anemia and beta-thalassemia has been associated with the co-inheritance of genes for hereditary persistence of fetal hemoglobin. A mouse locus has been identified that influences tumour number in mice with the multiple intestinal neoplasia gene. Understanding of the genetic interactions that determine phenotype in apparently monogenic diseases should lead to clarification of the role of different genes in polygenic diseases with complex inheritance patterns, as well as enhancing the ability to predict the outcome of a disease mutation. PMID- 8723678 TI - CDKN2A (p16INK4A) somatic and germline mutations. AB - The cell cycle is composed of a series of steps that can be negatively or positively regulated by various factors. A group of low-molecular-weight proteins have recently been identified that specifically inhibit the function of cyclin dependent kinases in mammalian cells. Inactivation of the CDKN2A gene (also known as p16INK4A and MTS1) attracted considerable interest after it was mapped to 9p21, a locus for familial melanoma. In an effort to standardize the information regarding human CDKN2A mutations detected in cancers, a database with information of 146 point mutations has been created. Cancer type, origin of cells, specific mutation, amino acid change, literature citation, and other data are provided for each mutation entry. Studies of biochemical and biological functions of both wild type and mutant proteins are central to our understanding of the role of p16INK4a mutations in tumorigenesis, a summary of these studies is also included in the present update. PMID- 8723679 TI - Characterization of large CTG repeat expansions in myotonic dystrophy alleles using PCR. AB - The CTG trinucleotide repeat expansions that are associated with myotonic dystrophy can be up to several thousand repeat units in length. We have developed a PCR protocol that has the potential to amplify mutant alleles with very large numbers of CTG repeats. The amplification uses the rTth DNA polymerase, XL system for long PCR targets together with primers which do not closely flank the repeat region and partial substitution of 7-deaza-dGTP for dGTP. Alleles containing up to approximately 800 CTG repeats were detected directly in agarose gels stained with ethidium bromide. Larger CTG repeat expansions required Southern blot transfer and detection with a repeat sequence probe; using this method, alleles containing up to approximately 2700 CTG repeats were detected. The PCR-based method described here was comparable to previous Southern blots of EcoRI restriction digested genomic DNA in both the approximate size and heterogeneity of mutant alleles detected, but provided more precise sizes of the CTG repeat expansions than the restriction digest approach. This PCR protocol could potentially simplify current mutation detection protocols in the molecular diagnosis of myotonic dystrophy, and facilitate molecular studies of the disease. PMID- 8723680 TI - Characterization of two arylsulfatase A missense mutations D335V and T274M causing late infantile metachromatic leukodystrophy. AB - Metachromatic leukodystrophy is a lysosomal storage disorder caused by the deficiency of arylsulfatase A. We describe a novel missense mutation in exon 6 causing the substitution of Asp335 by Val. In transient transfections no enzyme activity could be expressed from the arylsulfatase A cDNA carrying this mutation. Examination of the effects of the mutation in cells stably overexpressing the mutant enzyme revealed, that the mutant enzyme is catalytically inactive and degraded in an early biosynthetic compartment. We have also investigated the effects of a previously identified mutation (T274M). The specific catalytic activity of the Met274 substituted arylsulfatase is reduced to about 35% of the normal enzyme when measured with an artificial substrate. Most of this enzyme is also degraded in an early biosynthetic compartment. PMID- 8723681 TI - Mutation in the carboxy-terminal propeptide of the Pro alpha 1(I) chain of type I collagen in a child with severe osteogenesis imperfecta (OI type III): possible implications for protein folding. AB - A young girl presented with severe type III osteogenesis imperfecta; her otherwise healthy mother also had a mild connective tissue disorder with blue sclerae and recurrent joint dislocations. Skin fibroblast cultures from the child produced both normal and post-translationally over-modified type I collagen. The mutant collagen was poorly secreted but had normal thermal stability. Cyanogen bromide peptide maps of the abnormal protein indicated a C-terminal mutation. The mother's cells produced only normal-appearing collagens. Mismatch analysis and extensive sequencing of cDNAs covering the suspect region did not reveal any potentially causal changes in the triple helical domains of either the alpha 1(I) or alpha 2(I) chains. However, examination of the C-propeptide sequences revealed two heterozygous single base changes in the child. One, an A->C changing threonine to proline at residue 29 of the alpha 2(I) C-propeptide was also present in the mother and maternal grandfather but not in 50 unrelated control individuals. The second, a T->C altered the last amino acid residue of the alpha 1(I) C-propeptide from leucine to proline and had occurred de novo in the affected child. This mutation highlights the importance of the C-propeptides in molecular assembly but it is not clear how such an extreme mutation causes the delay in triple helix formation indicated by the extensive over-modification and reduced secretion of the mutant type I collagen. It may inhibit intrachain disulfide bonding or possibly affect the association of the procollagen chain with an intracellular "chaperone" protein that normally assists the assembly of trimeric procollagen molecules. PMID- 8723682 TI - Mutation of the hMSH2 gene in two families with hereditary nonpolyposis colorectal cancer. AB - We examined 18 unrelated individuals who have colorectal cancer or cancers associated with the HNPCC syndrome and have a family history of cancer for mutations in exon 13 of the hMSH2 gene. Two of the 18 individuals had the same previously unreported single-base deletion in codon 705 of hMSH2, resulting in a frame-shift mutation. Two other individuals had a T-to-C base change in the intron sequence at -6 position of the splice acceptor site at the 5' end of exon 13 which has previously been reported to be a polymorphism found in normal individuals (Leach et al., 1993). Nucleotide sequence changes were not detected in the remaining 14 individuals. We examined DNA from additional family members of the two subjects with codon 705 mutations. Of these, two individuals were identified with the mutation who were older than 50 years and who are apparently cancer free. The occurrence of breast cancer in both families (including one individual with a confirmed codon 705 mutation) suggests that breast cancer may be a part of the HNPCC syndrome. This is the first study to describe mutations in the hMSH2 gene in families that do not fit the definition of HNPCC. Because both families with the mutations at codon 705 failed to meet the Amsterdam criteria for HNPCC (Vasen et al., Dis Colon Rectum 34:424-425, 1991), our findings suggest that these criteria should now be reconsidered for purposes of diagnosis of HNPCC. PMID- 8723683 TI - Mutations in the BRCA1 gene in Japanese breast cancer patients. AB - Predisposing germline mutations in the BRCA1 gene were identified recently in families with 17 q-linked breast and ovarian cancers. Using single-strand conformation polymorphism (SSCP) analysis, we examined primary breast cancers for mutations in coding exons of BRCA1 in a panel of 103 patients, of whom all either represented early-onset cases (< 35 of age), were members of multiply-affected families, and/or had developed bilateral breast cancers. Mutations were detected in tumors from four patients, all of whom had developed breast cancers bilaterally: a frame-shift due to a 2-bp deletion at codon 797; a nonsense mutation at codon 1214; and two missense mutations, one at codon 271 leading to Val-->Met substitution, and the other at codon 1150 leading to Pro-->Ser substitution. In each case the same mutation was present in constitutional DNA. The mean age of onset was 49 years among the Japanese carriers of BRCA1 mutations identified in this study, in contrast to the mean age of 35 observed among carriers of BRCA1 mutations in a similar U.S. study (Futreal et al., 1994). The evidence reported here supports a rather limited role of BRCA1 in breast carcinogenesis. PMID- 8723684 TI - Phenotypic expression in double heterozygotes for familial hypercholesterolemia and familial defective apolipoprotein B-100. AB - Variability in the expression of monogenic lipid disorders may be observed in patients carrying the same DNA mutation, suggesting possible genetic or environmental interactions. Our objective was to investigate the genotype phenotype relationships in two unrelated French patients with an aggravated expression of a dominantly inherited hypercholesterolemia. In probands, segregation analysis complemented by DNA sequencing identified heterozygous defective alleles and mutations on two nonallelic loci for two monogenic lipid disorders: familial hypercholesterolemia at the low density lipoprotein (LDL) receptor locus and familial defective apolipoprotein B-100 at the locus encoding its ligand, apolipoprotein B-100. The LDL-receptor missense mutations had been reported in French Canadians. The apolipoprotein B mutation was the Arg3500Gln founder mutation in Northern Europe. Probands had an unusual phenotype of aggravated hypercholesterolemia that was complicated with premature coronary arterial disease, although remaining responsive to lipid-lowering drugs. This phenotype was distinct from that observed in their heterozygous relatives and distinct from those observed in FH or FDB homozygotes. These cases refer to a new class of patients with digenic lipid disorders, defined by specific clinical features that result from the combined effects of two independent loci. Moreover, the observed phenotype of aggravated hypercholesterolemia gives further evidence that receptor and ligand play distinct roles in regulating LDL metabolism. Although uncommon, these cases give insight into the molecular mechanisms that underly the clinical variability of inherited hypercholesterolemia. PMID- 8723685 TI - Mutation detection by solid phase primer extension. AB - A mutation analysis method based upon a wild-type DNA sequence is presented. Oligonucleotides were utilized for primer extension by T7 DNA polymerase to discriminate between wild-type and mutant sequences in two solid phase approaches. 1. Oligonucleotides were annealed to an immobilized template, extended with fluorescent dideoxynucleotides (ddNTPs), and analyzed on an automated fluorescent DNA sequencer. The oligonucleotide length identified the known mutation site, and the fluorescence emission of the ddNTP identified the mutation. 2. Template DNA was annealed to an oligonucleotide array, extended with alpha-32P dNTPs, and analyzed with a Phosphor Imager. The grid position of the oligonucleotide identified the mutation site and the extended base identified the mutation. PMID- 8723686 TI - Pelizaeus-Merzbacher disease: a novel mutation in the 5'-untranslated region of the proteolipid protein gene. PMID- 8723687 TI - A novel combination of mitochondrial tRNA and ND1 gene mutations in a syndrome with MELAS, cardiomyopathy, and diabetes mellitus. PMID- 8723688 TI - Two novel frameshift mutations causing premature stop codons in a patient with the severe form of Maroteaux-Lamy syndrome. PMID- 8723689 TI - Deletion of Gly723 in the insulin receptor substrate-1 of a patient with noninsulin-dependent diabetes mellitus. PMID- 8723690 TI - Six novel mutations in the fumarylacetoacetate hydrolase gene of patients with hereditary tyrosinemia type I. PMID- 8723691 TI - Estimation of carrier frequency of a 2.7 kb deletion allele of the P gene associated with OCA2 in African-Americans. PMID- 8723692 TI - Identification of a donor splice site mutation leading to loss of p22-phox exon 5 in autosomal chronic granulomatous disease. PMID- 8723693 TI - A novel mutation in exon 12 (Y569C) of the CFTR gene identified in a patient of Croatian origin. PMID- 8723694 TI - A donor splice site mutation (1811 + 1G-->C) in intron 11 of the CFTR gene identified in a patient of Macedonian origin. PMID- 8723695 TI - Identification of three novel mutations in the cystic fibrosis transmembrane conductance regulator gene in Argentinian CF patients. PMID- 8723696 TI - Identification of a novel PAX6 gene mutation in an aniridia patient. PMID- 8723697 TI - Identification of a 4 bp deletion (1560del4) in po gene in a family with severe Charcot-Marie-Tooth disease. PMID- 8723698 TI - Simple detection of a (Finnish) hereditary tyrosinemia type 1 mutation. PMID- 8723699 TI - Hippocampal afferents to the rat prefrontal cortex: synaptic targets and relation to dopamine terminals. AB - Afferents to the prefrontal cortex (PFC) from the hippocampal formation and from midbrain dopamine (DA) neurons have been implicated in the cognitive and adaptive functions of this cortical region. In the present study, we investigated the ultrastructure and synaptic targets of hippocampal terminals, as well as their relation to DA terminals within the PFC of adult rats. Hippocampal afferents were labeled either by anterograde transport of wheat germ agglutinin-horseradish peroxidase (WGA-HRP) from the ventral hippocampal formation or by anterograde degeneration following fimbria lesion. Hippocampal terminals in the PFC, identified by either method, formed primarily asymmetric axospinous synapses, with a small percentage forming asymmetric axodendritic synapses. Dopamine terminals in the PFC were identified by peroxidase immunocytochemistry for either tyrosine hydroxylase or DA and formed primarily symmetric synapses onto dendritic spines and small caliber dendritic shafts. Spines that received symmetric synaptic contact from DA terminals invariably also received an asymmetric synapse from an unlabeled terminal, forming a triadic complex. Hippocampal and DA terminals in the PFC were not often observed in the same area of the neuropil, and no examples of convergence of hippocampal and DA terminals onto common postsynaptic targets were observed. Further analysis revealed that spines receiving synaptic contact from hippocampal terminals did not receive additional synaptic contact from any other source. However, when localized to the same area of the neuropil, hippocampal and DA terminals were often in direct apposition to one another, without forming axo-axonic synapses. These results suggest that 1) hippocampal terminals primarily form excitatory synapses onto spiny pyramidal neurons, 2) hippocampal afferents are unlikely to be synaptically modulated by DA or non-DA terminals at the level of the dendritic spine, and 3) appositions between hippocampal and DA terminals may facilitate presynaptic interactions between these afferents to the PFC. PMID- 8723700 TI - Distribution of nitric oxide synthase-containing ganglionic neuronal somata and postganglionic fibers in the rat kidney. AB - Nitric oxide synthase (NOS)-immunoreactive neurons were identified in the rat kidney by using an antibody against type Ia NOS and the avidin-biotin complex immunoperoxidase method in whole kidneys examined in 100 microns serial sections. The histochemical method for demonstration of the nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) was also used to characterize NOS containing neurons. All somata showing NOS immunoreactivity also displayed NADPH d activity. The greatest number of neuronal somata were observed in groups at the wall of the renal pelvis and in the angular space formed by the pole of the renal parenchyma and renal pelvic wall. They were also seen at the renal hilus close to the renal artery and along the interlobar vasculature. The size of the neuronal somata in the 35-day-old rat ranged from 13.6 to 34.8 microns, with a mean size of 21.52 +/- 4.81 microns. Seventy percent, however, ranged in size from 17.8 to 26.8 microns. The shape of the neuronal somata also varied, with the majority having an ovoid or round shape. The distribution of the postganglionic fibers was investigated by means of the camera lucida. Postganglionic fibers projected into the wall of the renal pelvis and/or to the interlobar arteries extending to the arcuate arteries and to the beginning of the afferent arterioles. The NOS immunoreactive neurons may have a vasodilator and relaxing function on the renal pelvic wall and vasculature. In addition, the presence of NOS-containing nerve fibers in nerve bundles, which are known to have predominantly vasomotor and sensory fibers, suggest that they may have a possible modulatory role on renal neural function. PMID- 8723701 TI - Comparative ontogenic profile of cholinergic markers, including nicotinic and muscarinic receptors, in the rat brain. AB - The ontogenic profiles of several cholinergic markers were assessed in the rat brain by using quantitative in vitro receptor autoradiography. Brain sections from animals at different stages of development were processed with [3H]AH5183 (vesamicol; vesicular acetylcholine transport sites), [3H]N-methylcarbamylcholine (alpha(4)beta(2) nicotinic receptor sites), [3H]hemicholinium-3 (high-affinity choline uptake sites), [3H]3-quinuclidinyl benzilate (total population of muscarinic receptor sites), [3H]4-DAMP (muscarinic M1/M3 receptor sites), [3H]pirenzepine (muscarinic M1 receptor sites), and [3H]AF-DX 116 and [3H]AF-DX 384 (muscarinic M2 receptor sites) as radiolabeled probes. The results revealed that, by the end of the prenatal period (embryonic day 20), the densities of nicotinic receptor and vesicular acetylcholine transport sites already represented a considerable proportion of those observed in adulthood (postnatal day 60) in different laminae of the frontal, parietal, and occipital cortices, in the layers of Ammon's horn fields and the dentate gyrus of the hippocampal formation, as well as in the amygdaloid body, the olfactory tubercle, and the striatum. In contrast, at that stage, the densities of total muscarinic, M1/M3, M1, and possibly M2 receptor and high-affinity choline uptake sites represent only a small proportion of levels seen in the adult. Differences were also observed in the postnatal ontogenic profiles of nicotinic, muscarinic, vesamicol, and high-affinity choline uptake sites. For example, between postnatal weeks 3 and 5, the levels of M1/M3 and M1 sites were at least as high as in the adult, whereas M2 and high-affinity choline uptake site densities appeared to be delayed and to reach adult values only after postnatal week 5. With regard to cholinergic innervation in the developing rat brain, the present findings suggest a temporal establishment of several components of the cholinergic systems. The first components are the vesicular acetylcholine transporter and nicotinic sites; these are followed by M1/M3 and M1 sites and, finally, by M2 and high-affinity choline uptake sites. PMID- 8723702 TI - Distinct populations of cells in the adult dentate gyrus undergo mitosis or apoptosis in response to adrenalectomy. AB - Granule neurons of the rat dentate gyrus are born in adulthood as well as during development. Apoptotic cell death also occurs normally in this population throughout the life of the rat. Removal of adrenal steroids results in both increased production and increased degeneration of dentate gyrus granule cells. In order to determine whether the age of a cell affects its response to adrenalectomy (ADX), the numbers of dentate gyrus cells of different ages were assessed following ADX or sham operation. Older cells, i.e., those labeled with the thymidine analog bromodeoxyuridine (BrdU) on postnatal day (P) 6, were reduced in number following ADX on P60, and some had the morphologic characteristics of degenerating cells, indicating that significant numbers of mature cells die in response to ADX. In contrast, the number of younger cells, labeled with 3H-thymidine or BrdU in adulthood, 24 hours or 2 weeks before ADX, did not decrease, suggesting that these less mature cells do not die following ADX. An increase in the number of cells that are immunoreactive for proliferating cell nuclear antigen, a marker of dividing or recently mitotic cells, indicates that immature dentate gyrus cells divide following ADX. These results suggest that following ADX, mature cells born during the 1st postnatal week die, whereas immature cells divide. PMID- 8723703 TI - Plasticity of geniculocortical afferents following brief or prolonged monocular occlusion in the cat. AB - During a critical period in early life, physiological studies reveal that either prolonged or brief periods of monocular occlusion induce similar plastic changes in the primary visual cortex (area 17) of the cat, leading to a nearly complete loss of visual responses from the deprived eye (Hubel and Wiesel [1970], J. Physiol. (London) 160: 106-154). However, the recovery of function is markedly different in the two conditions, being complete and thorough only after a brief period of monocular occlusion (Movshon [1976] J. Physiol. (London) 261: 125-174). In search for anatomical correlates that distinguish between these two experimental conditions, we compared the geniculocortical connectivity in normal kittens with that following brief (4 days and 6-7 days) or prolonged (> 5 weeks) periods of occlusion of vision in one eye. Transneuronal labeling of the geniculocortical pathway revealed changes in both cases, and single afferent arbors reconstructed in serial sections were similarly shrunken after short or long periods of deprivation. However, a significant expansion of the geniculocortical afferents serving the nondeprived eye was evident mainly after prolonged deprivation. These findings provide further evidence for rapid, activity-dependent remodeling of afferents during development. PMID- 8723704 TI - Distribution of GABAA, GABAB, and glycine receptors in the central auditory system of the big brown bat, Eptesicus fuscus. AB - Quantitative autoradiographic techniques were used to compare the distribution of GABAA, GABAB, and glycine receptors in the subcortical auditory pathway of the big brown bat, Eptesicus fuscus. For GABAA receptors, the ligand used was 35S-t butylbicyclophosphorothionate (TBPS) for GABAB receptors, 3H-GABA was used as a ligand in the presence of isoguvacine to block binding to GABAA sites; for glycine, the ligand used was 3H-strychnine. In the subcortical auditory nuclei there appears to be at least a partial complementarity in the distribution of GABAA receptors labeled with 35S-TBPS and glycine receptors labeled with 3H strychnine, GABAA receptors were concentrated mainly in the inferior colliculus (IC) and medial geniculate nucleus, whereas glycine receptors were concentrated mainly in nuclei below the level of the IC. Within the IC, there was a graded spatial distribution of 35S-TBPS binding; the most dense labeling was in the dorsomedial region, but very sparse labeling was observed in the ventrolateral region. There was also a graded spatial distribution of 3H-strychnine binding. The most dense labeling was in the ventral and lateral regions and the weakest labeling was in the dorsomedial region. Thus, in the IC, the distribution of 35S TBPS was complementary to that of 3H-strychnine. GABAB receptors were distributed at a low level throughout the subcortical auditory nuclei, but were most prominent in the dorsomedial part of the IC. PMID- 8723706 TI - Light and electron microscopic immunohistochemical study of dopaminergic terminals in the striatal portion of the pigeon basal ganglia using antisera against tyrosine hydroxylase and dopamine. AB - A dopaminergic projection from the midbrain to the striatal portion of the basal ganglia is present in reptiles, birds, and mammals. Although the ultrastructure of these fibers and terminals within the striatum has been studied extensively in mammals, little information is available on the ultrastructure of this projection in nonmammals. In the present study, we used immunohistochemical labeling with antibodies against tyrosine hydroxylase (TH) or dopamine (DA) to study the dopaminergic input to the striatal portion of the basal ganglia in pigeons (i.e., lobus parolfactorius and paleostriatum augmentatum). At the light microscopic level, the anti-TH and anti-DA revealed a similar abundance and distribution of numerous labeled fine fibers and varicosities within the striatum. In contrast, the use of an antidopamine beta-hydroxylase antiserum (which labels only adrenergic and noradrenergic terminals) labeled very few striatal fibers, which were restricted to visceral striatum. These results demonstrate that anti-TH mainly labels dopaminergic terminals in the striatum. At the electron microscopic level, the anti-TH and anti-DA antisera labeled numerous axon terminals within the striatum (15-20% of all striatal terminals). These terminals tended to be small (with an average length of 0.6 microns) and flattened, and their vesicles tended to be small (35-60 nm in diameter) and pleomorphic. About 50% of the terminals were observed to make synaptic contacts in the planes of section examined, and nearly all of these synaptic contacts were symmetric. Both TH+ and DA+ terminals typically contacted dendritic shafts or the necks of dendritic spines, but a few contacted perikarya. No clear differences were observed between TH+ and DA+ terminals within medial striatum (whose neurons project to the nigra in birds) or between TH+ and DA+ terminals within lateral striatum (whose neurons project to the pallidum in birds). In addition, no differences were observed between medial and lateral striata in either TH+ or DA+ terminals. Thus, there is no evident difference in pigeons between striatonigral and striatopallidal neurons in their dopaminergic innervation. Our results also indicate that the abundance, ultrastructural characteristics, and postsynaptic targets of the midbrain dopaminergic input to the pigeon striatum are highly similar to those in mammals. This anatomical similarity is consistent with the pharmacologically demonstrable similarity in the role of the dopaminergic input to the striatum in birds and mammals. PMID- 8723705 TI - Eicosanoids and nitric oxide influence induction of reactive gliosis from spreading depression in microglia but not astrocytes. AB - Microglia and astrocytes are transformed into reactive glia (RG) by brain disease and normal function. Eicosanoids and nitric oxide (NO), two intercellular mediators, may influence gliosis. We investigated how drugs that alter production of these paracrine signals effect induction of glial reactivity from spreading depression. Unilateral (left) neocortical spreading depression was induced in 95 halothane anesthetized rats by intracortical injections of 0.5 M KCl, with or without drug treatment (five animals/group). Immunohistochemical staining (IS) intensity using the OX-42 and anti-glial fibrillary acidic protein (GFAP) antibodies determined reactivity in microglia and astrocytes, respectively. After 3 days, brains were processed for OX-42 and GFAP-IS and mean optical densities (OD) of IS were measured. Average OD's (for OX-42) and the log ratio (left/right) of OD's (OX-42 and GFAP) were compared to normal animals. Spreading depression induced significant log ratios for both OX-42- and GFAP-IS (P's < 0.01). However, dexamethasone (a glucocorticoid), nordihydroguaiaretic acid (a lipoxygenase inhibitor), and nitroprusside (a NO donor) prevented significant left sided and log ratio OD values for microglia (P's > 0.05). L-Name, a NO synthase inhibitor, caused significant increases in left and right OD's for microglia (P's < 0.05). Mepacrine, a phospholipase A2 inhibitor, Indomethacin, a cyclooxygenase inhibitor, and phenylephrine, an adrenergic agonist, did not prevent induction of significant OX-42 log ratios (P's < 0.01, 0.05, 0.01), and resulted in increases in left side OD's (P's < 0.01, 0.05, 0.05). Significant GFAP log ratios occurred after spreading depression in all drug groups, P's < 0.01. Thus, induction of reactivity in microglia is more sensitive to eicosanoids and NO than in astrocytes. PMID- 8723707 TI - Presynaptic and postsynaptic subcellular localization of substance P receptor immunoreactivity in the neostriatum of the rat and rhesus monkey (Macaca mulatta). AB - The substance P receptor (SPR) gene is expressed at high levels in basal ganglia, but the paucity of information about localization of the encoded receptor protein has limited our understanding of this peptide's involvement in cellular and subcellular mechanisms in this region. Morphological evidence in the rodent striatum indicates that SPRs are expressed in postsynaptic neuronal elements, while pharmacological studies suggest the existence of presynaptic SPRs in this structure. We have examined the issue of subcellular distribution of this receptor protein in rat and primate neostriatal tissue, employing an antiserum raised against SPR. Electron microscopic analysis revealed that SPR immunoreactivity is present in presynaptic and postsynaptic neuronal elements in both species. In agreement with earlier studies, SPR immunoreactivity was found predominantly in perikarya and dendrites of a small subset of striatal neurons, the large and medium-sized aspiny interneurons. In addition, a small but significant proportion of the immunoreaction product was localized in presynaptic profiles, both in axons and axon terminals. The majority of SPR immunoreactive boutons formed asymmetric synapses with dendrites and dendritic spines. The association of SPRs with asymmetric synapses provides a morphological substrate for peptidergic modulation of excitatory neurotransmission of extrastriatal origin. A minor proportion of immunolabeled axons established symmetric synaptic junctions with unlabeled dendrites. The presence of SPRs in these synapses suggests a presynaptic peptidergic modulation of intrinsic striatal transmitter systems. The observations in this study also indicate that SPR mediates a complex combination of postsynaptic and presynaptic effects on acetylcholine release in the mammalian striatum. PMID- 8723708 TI - Distribution and neurochemical character of substance P receptor (SPR) immunoreactive striatal neurons of the macaque monkey: accumulation of SP fibers and SPR neurons and dendrites in "striocapsules" encircling striosomes. AB - The striatal distribution of the substance P receptor (SPR) protein was examined in relation to its ligand, the neuro-peptide SP, as well as to the neurochemical and compartmental composition of the neostriatum in rhesus monkeys (Macaca mulatta) in immunohistochemical experiments. About 2% of striatal neurons, displaying varicose, virtually spine-free dendrites characteristic of large and medium-sized aspiny interneurons, expressed SPR immunoreactivity. SPR/choline acetyltransferase, SPR/somatostatin, SPR/GABA, SPR/calbindin D28k, and SPR/parvalbumin double immunolabeling experiments demonstrated that SPR-positive cells are either cholinergic or somatostatinergic. Comparison of SP and SPR immunoreactivities in double-labeled and adjacent single-labeled sections revealed compartment-specific match and mismatch between the densities of the peptide and receptor. A matching high density of SP fibers and SPR cells and dendrites was only observed in the rim of the striosome compartments. To our knowledge, this is the first evidence for an anatomical border comprised of dendritic processes that separate striatal compartments. We have termed these zones "striocapsules," because they encircle and encapsulate striosomal cell islands. In the striatal matrix, an abundance of SPR-labeled profiles was complemented with light SP staining. By contrast, in the core of the striosomes, SPR labeling was sparse and SP staining intense. SP-positive axon-like puncta frequently contacted SPR-positive dendrites in all striatal compartments. The SP receptor/ligand match indicates a sharp increase in the efficacy of SP action in the striocapsules, and suggests that the influence of SP might be heightened in this striatal subcompartment. PMID- 8723709 TI - Cerebellar granule cell differentiation in mutant and X-irradiated rodents revealed by the neural adhesion molecule TAG-1. AB - In the external granular layer of the cerebellum, the granule cell precursors express the transient axonal glycoprotein TAG-1, a molecule involved in adhesion and neurite outgrowth. Granule cells express TAG-1 transiently, just as they extend neurites before migrating over the radial glia. The present study aims to investigate whether the expression pattern of TAG-1 is altered when granule cells develop abnormally. We studied in vivo models in which Purkinje and/or granule cell defects occur during postnatal development. These include the cerebellar mutant mice staggerer and lurcher as well as rats irradiated during postnatal development. Neither alterations in Purkinje cell differentiation nor the related granule cell loss in the mouse mutants impairs the ability of the surviving granule cell precursors to express TAG-1. Also, early granule cell loss in the X irradiated rats do not disturb the TAG-1 expression phase in the patches of surviving granule cell precursors. Ectopic granule cells found in the adult cerebellum of X-irradiated rats do not bear the molecule, although they are located in the most superficial part of the molecular layer, occupied by the immunopositive cells a few days earlier. Thus, TAG-1 marks a very precise stage of granule cell differentiation, and the inward migration process itself is not required for the cessation of the expression. We postulate that TAG-1 may be involved in local differentiation steps restricted to the deep external granular layer such as parallel migratory routes or synchrony of axonal growth. PMID- 8723710 TI - Effect of dopaminergic drugs on the in vivo binding of [3H]WIN 35,428 to central dopamine transporters. AB - [11C]WIN 35,428 (also designated [11C]CFT) is now being used in several positron emission tomography (PET) centers to image dopamine (DA) transporter sites in the mammalian brain. Whether and to what extent in vivo WIN 35,428 binding is influenced by intra- and extrasynaptic dopamine levels are largely unknown. The purpose of the present study was to evaluate the effects of various drugs, known to affect DA levels and release, on the binding of [3H]WIN 35,428 to central DA transporters in the mouse brain. D-Amphetamine, which releases DA from neurons and blocks the DA transporter directly, inhibited striatal [3H]WIN 35,428 binding in dose-dependent manner. Similarly, alpha-methyl-DL-p-tyrosine, an inhibitor of tyrosine hydroxylase, blocked [3H]WIN 35,428 binding, possibly via competitive inhibition by the metabolite p-hydroxyamphetamine. Specific binding of [3H]WIN 35,428 was insensitive to changes in synaptic DA levels caused by pretreatment of the animals with high doses of D2 receptor agonists (apomorphine, bromocriptine), antagonists (haloperidol) or the inhibitor of dopaminergic neuron firing gamma butyrolactone (GBL). High doses (> 50 mg/kg) of L-DOPA (in combination with benserazide), however, reduced [3H]WIN 35,428 binding significantly, yet for a relatively short time (approximately 2.5 h). Chronic treatment with L-deprenyl elicited no changes in in vivo [3H]WIN 35,428 accumulation in the striatum. Neurotoxic damage of DA neurons caused by administration of high doses of amphetamine was detected in the striatum by a significant reduction in [3H]WIN 35,428 binding 7 days after cessation of amphetamine treatment. Thus, [3H]WIN 35,428 binding was only affected by neurotoxic loss of neurons, by administration of uptake inhibitors, or by some treatments which significantly elevate DA levels. Compounds which inhibit DA release or deplete DA acutely do not increase [3H]WIN 35,428 binding, suggesting that normal or "resting" levels of DA are not sufficient to alter [3H]WIN 35,428 binding in vivo. These findings are important for our understanding of the function and regulation of the DA transporter, as well as the in vivo binding of the radioligand [3H/11 C]WIN 35,428. Moreover, they will be important for the interpretation of PET studies in which [11C]WIN 35,428 is used to assess the integrity of dopaminergic neurons. PMID- 8723711 TI - Effects of dopamine on neurons of the lateral geniculate nucleus: an iontophoretic study. AB - In urethane anesthetized rats, the iontophoretic administration of dopamine (DA) induced an inhibition of flash-evoked activity in the majority of geniculate cells investigated. Excitatory effects of DA also were found in some neurons of the dorsal lateral geniculate nucleus. The observed excitatory effects of DA were blocked selectively by D2 receptor antagonists, and the majority of inhibitory effects could be blocked by D1 receptor antagonists. In some neurons, the D2 receptor antagonist also blocked the DA-induced inhibition. Nine of 33 neurons tested responded differently to DA according to the amount ejected: with lower iontophoretic currents they increased their rates of discharge, whereas higher DA ejecting currents resulted in a suppression of their activity. Iontophoretic administration of a D1 agonist (SKF 38393) for the most part induced a decrease in baseline activity, whereas the D2 agonist (quinpirole) frequently induced an increase. These effects of agonists were dose-dependent and reproducible. Effects of the D1 and D2 agonists could be reversed by the receptor-specific dopamine antagonists. Presumed local circuit interneurons appeared to be involved in mediation of some inhibitory effects of DA, since the D2-induced inhibitions could be abolished by the GABAA antagonist, bicuculline. The majority of cells also was affected by DA antagonists given alone; these cells' responses to light usually were of an inhibitory nature. The results show that like other monoamines, DA also is involved in certain aspects of visual processing at the level of the thalamus. PMID- 8723712 TI - Rabphilin-3A is transported with fast anterograde axonal transport and associated with synaptic vesicles. AB - Rab3a has been proposed to be involved in the process of exocytosis. It undergoes a membrane association/dissociation cycle which is dependent upon alteration in its GTP-GDP state. Rabphilin-3A is one of the most attractive candidates for a Rab3a effector molecule in synaptic vesicle exocytosis. It binds selectively to the GTP-bound form of Rab3a as well as of Rab3c. Fast axonal transport was studied by crushing spinal roots and sciatic nerves, resulting in accumulations of transported, organelle-bound substances. Rabphilin-3A was transported with fast anterograde transport, but < 10% of the proximal accumulation was detected in the recycling vesicle population. The pattern for accumulation of rabphilin-3A was similar to that of Rab3a accumulation, including the poor recycling. However, synaptophysin and SV2, two transmembrane synaptic vesicle proteins, accumulated in large amounts in many axons distal to the crushes, and the amounts of recycling synaptophysin and SV2, in percentages of anterograde accumulations, were about 70% and 60%, respectively. Double-labelling showed a high degree of colocalization between Rab3a and rabphilin-3A in spinal roots and sciatic nerves. The two proteins essentially colocalized in the nerve terminals of the spinal cord and in motor endplates. Immuno-EM studies demonstrated that rabphilin-3A was present on clear small synaptic vesicles in the proximal segment, but was absent from vesicles in the distal segment. Rabphilin-3A also associated with the synaptic vesicles in the presynaptic compartment. Results indicate that rabphilin 3A is associated with the membrane of synaptic vesicles during anterograde transport. Furthermore, as it colocalized in most, but not all, structures with Rab3a, results support the hypothesis that rabphilin-3A is interacting with Rab3a. PMID- 8723713 TI - Striatal c-fos levels do not correlate with haloperidol-induced behavioral supersensitivity. AB - Striatal c-fos levels and stereotyped behavior have been evaluated in chronically haloperidol-treated rats which received subsequent subacute dopamine (DA) agonist treatment to investigate the possible relationship between striatal c-fos and behavioral supersensitivity. Haloperidol treatment (1 mg/kg/day for 21 days) increased apomorphine-induced stereotypies but did not modify striatal c-fos levels. The subacute administration of the DA D-1 agonist SKF38393 (10 mg/kg/day for 5 days) and the combination of the D-1 agonist with the D-2 agonist quinpirole (1 mg/kg/day for 5 days) attenuated apomorphine-induced stereotypies after haloperidol pretreatment. The administration of quinpirole alone, however, did not modify the response to haloperidol. All DA agonists significantly increased c-fos levels after apomorphine injection. The dissociation between haloperidol-induced behavioral supersensitivity and striatal c-fos levels observed in this study suggests that mechanisms different from striatal c-fos induction might be involved, and that striatal c-fos levels are not a good marker of behavioral supersensitivity expression. PMID- 8723714 TI - Distribution of preproenkephalin, preprotachykinin A, and preprodynorphin mRNAs in the rat nucleus accumbens: effect of repeated administration of nicotine. AB - The effects of a repeated treatment with nicotine on the expression of mRNAs encoding preproenkephalin (PPE), preprotachykinin-A (PPT-A), and preprodynorphin (PPDYN) were examined by in situ hybridization histochemistry in various subregions of the nucleus accumbens (Acb). In saline-treated rats, optical density measurements on autoradiographic films showed marked anteroposterior decreasing gradients for PPE and PPT-A mRNAs in the rostral pole and the core, in the cone, and in the ventral shell of the Acb, whereas a lower anteroposterior gradient was observed for PPDYN mRNA signals. The intensity of the three mRNA signals also varied according to Acb subregion. However, analysis of percentages of prepropeptide mRNA-containing neurons as compared to total neurons showed, in the rostral pole, the core, and the cone, a similar percentage of PPE mRNA (around 45%)- and PPT-A mRNA (around 40%)-expressing neurons. The ventral shell can be distinguished from the other subregions by a lower percentage of PPE mRNA (35.8%)- and PPT-A mRNA (30.6%)-expressing neurons. The percentage of PPDYN mRNA containing neurons, by contrast, was similar (around 37%) in the core, the cone, and the ventral shell. Repeated nicotine administration increases the PPE mRNA level in the rostral pole and the anterior third of the core without any change in PPT-A and PPDYN mRNA levels in the various Acb subregions examined. The PPE mRNA increase does not support an effect mediated through an interaction of nicotine with DA neurons. The effect could be linked to a nicotine activation of other afferents to the anterior Acb and/or to a direct nicotine stimulation of PPE mRNA neurons. PMID- 8723715 TI - Striatal efferents preferentially innervate neurons in the ventral pallidum containing GABAA receptor alpha 1 subunit-like immunoreactivity. AB - The gamma-aminobutyric acid-A receptor consists of several subunits. In this immunohistochemical study we investigated the regional distribution of the alpha 1 and alpha 2 subunits with subunit-specific antibodies in the ventral pallidum, and compared the staining patterns to those of substance P (SP). alpha 1 subunit antigenic sites were found to be localized to pallidal neurons, varicosities, and varicose fibers. alpha 1 immunopositive fibers mainly appeared "tubulus-like" due to the intense staining of the membranes of the long pallidal dendrites. Double labelling of alpha 1 subunit and substance P revealed that alpha 1 subunit-like immunoreactive (IR) dendrites and somata of the pallidal neurons were often invested by SP-IR striatal efferents. Subcellularly, the dendritic and somatic membranes of pallidal neurons were strongly immunopositive for the alpha 1 subunit, whereas only a few axon terminals exhibited alpha 1-IR. alpha 2-IR was restricted to a low number of ventral pallidal neurons. The distributional patterns obtained for the alpha 1 and alpha 2 subunits suggest that striatal efferent neurons directly influence pallidal neurons displaying a distinct GABAA subunit composition, which may be of pharmacological importance since the alpha 1 beta x gamma 2-subunits containing receptors have mainly a benzodiazepine type I pharmacology. PMID- 8723716 TI - Autoradiographic localization of extrastriatal D2-dopamine receptors in the human brain using [125I]epidepride. AB - Epidepride is a benzamide with high affinity for central D2- and D3-dopamine receptors. The anatomical distribution of [125I]epidepride binding was examined by autoradiography, using postmortem human whole-hemisphere cryosections. The density of [125I]epidepride binding sites was high in caudate nucleus and putamen. [125I]epidepride also labeled receptors in extrastriatal region such as in the pallidum, some thalamic nuclei, the neocortex, and the substantia nigra. The neocortical binding was heterogeneously distributed. In most cortical regions, binding sites were located in superficial layers (I-II). However, in basal levels of the occipital cortex, [125I]epidepride binding was located in a deeper layer, probably corresponding to layer V. Competition studies indicated that most of the [125I]epidepride binding represented predominantly D2-dopamine receptors, in striatal as well as in extrastriatal regions. The presence of extrastriatal D2-dopamine receptor populations is of particular interest for research on schizophrenia and antipsychotic drug action. PMID- 8723717 TI - Mapping the human Y chromosome by fingerprinting cosmid clones. AB - We have used Y-specific cosmid clones in a random fingerprinting approach to build contigs on the human Y chromosome. Clones derived from two libraries have been analyzed. The construction of one library is described here, the second was the Y chromosome-specific library LLOYNCO3 "M" (Lawrence Livermore National Laboratory). To date, we have fingerprinted 4430 cosmids: 377 contigs have been constructed containing from 2 to 39 clones. Along with the singletons, we estimate that we have covered 72.5% of the euchomatic portion of the Y chromosome with fingerprinted clones. Sequence tagged sites are being used to anchor cosmids and contigs onto the YAC framework. PMID- 8723718 TI - Dominantly and recessively inherited cornea plana congenita map to the same small region of chromosome 12. AB - Cornea plana congenita occurs in a mild autosomal dominant (CNA1) and a more severe autosomal recessive (CNA2) form. We recently assigned a CNA2 locus to a region on chromosome 12 by linkage analysis and excluded linkage to that locus in two Finnish CNA1 families. Here we describe a Cuban pedigree in which 14 members are affected with dominantly inherited cornea plana. By linkage analysis this phenotype was mapped to the immediate vicinity of markers D12S82 and D12S351 on 12q, that is, precisely the same small region (3 cM or less) to which CNA2 previously had been assigned. Our results support the existence of at least three genetically distinct forms of cornea plana. It remains to be determined whether recessive and dominant cornea plana are caused by different mutations of a single gene or whether the region in 12q harbors two or more genes whose mutations cause corneal maldevelopment. PMID- 8723719 TI - Mapping the RP10 locus for autosomal dominant retinitis pigmentosa on 7q: refined genetic positioning and localization within a well-defined YAC contig. AB - Retinitis pigmentosa is a genetically heterogeneous disease that has autosomal dominant, autosomal recessive and X-linked forms. Autosomal dominant retinitis pigmentosa (adRP) has thus far been associated with eight distinct loci, including the rhodopsin and peripherin/RDS genes as well as unidentified genes on chromosomes 7p, 7q, 8q, 17p, 17q, and 19q. The RP10 locus for adRP on chromosome 7q was first mapped in a Spanish family; later, an unrelated American family was identified that also showed linkage to 7q. By combining the linkage results from both families, we are able to assign the disease gene to a 5-cM interval on 7q. Based on extensive physical mapping of this region, the genetic interval is now fully contained within a approximately 5-Mb segment on a well-defined YAC contig. These studies significantly reduce the size of the RP10 critical region, exclude a number of possible candidate genes, and provide the necessary cloned DNA for the positional cloning of the RP10 gene. PMID- 8723720 TI - A palindromic structure in the pericentromeric region of various human chromosomes. AB - The primate-specific multisequence family chAB4 is represented with approximately 40 copies within the haploid human genome. Former analyis revealed that unusually long repetition units ( > 35 kb) are distributed to at least eight different chromosomal loci. Remarkably varying copy-numbers within the genomes of closely related primate species as well as the existence of human specific subfamilies, which most probably arose by frequent sequence exchanges, demonstrate that chAB4 is an unstable genomic element, at least in an evolutionary sense. To analyze the chAB4 basic unit in more detail we established a cosmid contig and found it to be organized as inverted duplications of approximately 90 kb flanking a noninverted core sequence of approximately 60 kb. FISH as well as the analysis of chromosome specific hybrid cell lines revealed a chromosomal localization of chAB4 on chromosomes 1, 3, 4, 9, Y, and the pericentromeric region of all acrocentrics. Furthermore, we can detect chAB4 sequences together with alpha satellites, beta satellites, and satellite III sequences within a single chromosome 22-specific YAC clone, indicating that chAB4 is located in close proximity to the centromere, at least on the acrocentrics. Hence, chAB4 represents an unstable genomic structure that is located just in the chromosomal region that is very often involved in translocation processes. PMID- 8723721 TI - Genetic mapping in Xiphophorus hybrid fish: assignment of 43 AP-PCR/RAPD and isozyme markers to multipoint linkage groups. AB - The combined use of the arbitrarily primed polymerase chain reaction [AP-PCR, also known as random amplification of polymorphic DNA (RAPD)] and isozyme mapping resulted in the production of 87 potential marker loci, enabling an overall expansion within the genetic map of the fish genus Xiphophorus. Use of DNA sequencing-style acrylamide gels and carefully controlled conditions of amplification and silver staining allowed exceptional resolution and reproducibility of AP-PCR/RAPD generated markers. Linkage analysis of AP-PCR/RAPD and isozyme markers resulted in the addition of 16 new markers to Xiphophorus linkage groups (LGs) I, II, III, V, IX, X, XII, and XIV. Addition of 5 AP PCR/RAPD markers to linkage group U6 containing the Tailspot pigment pattern locus (P) and designation of eight new unassigned linkage groups with 22 markers was also accomplished. Genetic linkage data allowed inference of the existence of a novel pigment pattern modifier locus. Expansion of the Xiphophorus gene map by linkage analysis of AP-PCR/RAPD markers in conjunction with isozyme polymorphisms should lead to the rapid saturation of genetic linkage groups such as LG V, which will probably be instrumental to cloning the Diff tumor suppressor gene locus. PMID- 8723722 TI - Toward the construction of integrated physical and genetic maps of the mouse genome using interspersed repetitive sequence PCR (IRS-PCR) genomics. AB - Using two recently developed techniques, IRS-PCR YAC walking and IRS-PCR genotyping, a framework-integrated physical and genetic map of the mouse genome was constructed. The map consists of 821 contigs, containing 7746 YAC clones originating from three different YAC libraries. Three hundred eighty of the contigs have been anchored to the genetic map. Approximately 16% of the physical length of the mouse genome is estimated to be represented. PMID- 8723723 TI - A tandem duplication within the fibrillin 1 gene is associated with the mouse tight skin mutation. AB - Mice carrying the Tight skin (Tsk) mutation have thickened skin and visceral fibrosis resulting from an accumulation of extracellular matrix molecules. These and other connective tissue abnormalities have made Tskl + mice models for scleroderma, hereditary emphysema, and myocardial hypertrophy. Previously we localized Tsk to mouse chromosome 2 in a region syntenic with human chromosome 15. The microfibrillar glycoprotein gene, fibrillin 1 (FBN1), on human chromosome 15q, provided a candidate for the Tsk mutation. We now demonstrate that the Tsk chromosome harbors a 30- to 40-kb genomic duplication within the Fbn1 gene that results in a larger than normal in-frame Fbn1 transcript. These findings provide hypotheses to explain some of the phenotypic characteristics of Tskl + mice and the lethality of Tsk/Tsk embryos. PMID- 8723725 TI - An efficient method for isolating putative promoters and 5'-transcribed sequences from large genomic clones. AB - Efficient strategies to isolate promoters and flanking exons from large genomic clones would facilitate the assembly of transcription units, complement existing techniques to isolate expressed sequences, and provide 5' regulatory elements. We have developed a rapid and simple method to isolate promoters from large mammalian genomic DNA clones by exploiting the abundance of binding sites for the ubiquitous transcription factor Sp1 in gene promoters. Using this method, putative promoter sequences with Sp1-binding sites are enriched approximately 100 fold from fragmented P1 clone DNA. Based on the abundance of Sp1-binding motifs in promoters, we predict that a significant subset of vertebrate promoters could be isolated by this method. PMID- 8723724 TI - A gene-rich cluster between the CD4 and triosephosphate isomerase genes at human chromosome 12p13. AB - The genomic sequence of the human CD4 gene and its neighboring region, located at chromosome 12p13, was generated using the large-scale shotgun sequencing strategy. A total of 117 kb of genomic sequence and approximately 11 kb of cDNA sequence were obtained. Six genes, including CD4, triosephosphate isomerase, B3 subunit of G proteins (GNB3), and ubiquitin isopeptidase T (ISOT), with known functions, and two new genes with unknown functions were identified. Using a battery of strategies, the exon/intron boundaries, splice variants, and tissue expression patterns of the genes were determined. Various computer software was utilized for analyses of the DNA and amino acid sequences. The results of the analyses and sequence-based strategies for gene identification are discussed. PMID- 8723726 TI - Single nucleotide primer extension: quantitative range, variability, and multiplex analysis. AB - The quantitative measurement of transcription products from homologous alleles at a diploid locus has broad application for the study of mammalian gene expression. Single nucleotide primer extension (SNuPE) analysis is a simple and sensitive method for allelic transcript discrimination requiring only 1 bp difference between alleles. In this study the effective range, experimental variation, and the influences of poly(dT)-primed and gene-specific reverse transcriptions are characterized. The ability to analyze several genes from a single reverse transcription reaction is assessed as well. For the genes examined, the maximum range of detection is reached when the minor transcript represents 1/250 of the major allele. Relatively little error is seen within or between assays and linearity of response is maintained over an approximately thousandfold range. PMID- 8723730 TI - Characterization of microsomal cytochrome P450 enzymes involved in the oxidation of xenobiotic chemicals in human fetal liver and adult lungs. AB - Levels and catalytic activities of cytochrome P450 (P450) enzymes involved in the oxidation of drugs and carcinogens were determined in human adult lungs and fetal livers and compared with those in microsomes from adult livers. P450s immunoreactive with anti-human P4501A1 and anti-human P4503A antibodies were detected in fetal liver microsomes by immunoblotting analysis, and P450s related P4501A1, 2A6, 2C9, 2E1, and 3A4 were determined in adult lung microsomes; all of these P450 enzymes were detected in much higher amounts in adult liver microsomes except that P4501A2 was only the 1A subfamily of P450 found in adult livers. Drug oxidation activities with the substrates ethoxyresorufin, coumarin, 7 ethoxycoumarin, bufuralol, and testosterone were determined in these microsomes, and we found that none of the activities were higher in microsomes of adult lungs and fetal livers than in adult livers. Activation of procarcinogens to reactive metabolites that induce umu gene expression in Salmonella typhimurium TA1535/pSK1002 or NM2009 was also examined and it was found that activities with (+)- and (-)-enantiomers of 7,8-dihydroxy-7,8-dihydrobenzo[a]pyrene were higher in fetal liver microsomes than adult lung or liver microsomes. The adult liver and lung activities for these two procarcinogens were similar on the basis of microsomal protein contents despite the fact that p450 contents are higher in liver than lung microsomes. alpha-Naphthoflavone, a known inhibitor of P4501A related activities, did not affect these procarcinogen activation in fetal liver microsomes. Fetal liver microsomes catalyzed activation of aflatoxin B1 and sterigmatocystin, two procarcinogens known to be activated by P4503A4/7 in humans, although activation of carcinogenic arylamines that are good substrates for P4501A2 was much lower in microsomes of fetal livers and adult lungs than in adult livers. These results suggest that in human fetal livers at least two P450 enzymes, a form of P450 that is immunoreactive P4501A1 and P4503A7, are actually expressed and these enzymes are suggested as being involved in the activation of the (+)- and (-)-enantiomers of 7,8-dihydroxy-7,8-dihydrobenzo[a]pyrene and the carcinogenic mycotoxins, respectively. The exact nature of the former enzyme in fetal livers is unknown. In adult human lungs, several P450 enzymes are expressed, although the precise roles of these enzymes in the oxidation of xenobiotics were not determined due to the low level of expression of these P450s. PMID- 8723729 TI - Protein-reactive metabolites of carbamazepine in mouse liver microsomes. AB - The character of reactive metabolites formed from carbamazepine (CBZ) was sought in incubations of [14C]CBZ in hepatic microsomes prepared from adult female mice of a strain (SWV/Fnn) susceptible to CBZ-induced teratogenicity. The formation of radio-labeled protein adducts was used as an index of reactive metabolite exposure. A dependence on cytochrome P450 was shown by a requirement for NADPH and inhibition by carbon monoxide, 1-aminobenzotriazole, piperonyl butoxide, and stiripentol. The addition of ascorbic acid, caffeic acid, N-acetylcysteine, and glutathione decreased the rate of binding of the radiolabel from [14C]CBZ to microsomal protein by more than 50%. The addition of glutathione transferases diminished protein adduct formation beyond that seen with glutathione alone. Evidence for the formation of an arene oxide was sought through the use of inhibitors of epoxide hydrolases, including cyclohexene oxide, chalcone oxides (with the addition of cytosol as appropriate), and by the addition of recombinant human soluble and microsomal epoxide hydrolases and recombinant rat microsomal epoxide hydrolase. The microsomal epoxide hydrolases decreased the velocity of 14C-labeled protein adduct formation by approximately 23%, whereas inhibitors had no effect, most likely because of the low native activity of microsomal epoxide hydrolase in mice. Both DT-diaphorase and catechol-O-methyltransferase diminished 14C-labeled protein adduct formation by 54% and 45%, respectively. The data suggest that the major reactive metabolites formed from CBZ by adult female SWV/Fnn liver microsomes are quinones and arene oxides. PMID- 8723731 TI - Characterization of the effects of tebufelone on hepatic cytochromes P450 in the beagle dog. AB - Tebufelone (1-[3,5-bis(1,1-dimethylethyl)-4-hydroxy-phenyl]-hex-5-yne-1-one) is an investigational ditertiary butylphenol nonsteroidal anti-inflammatory drug. The purpose of the present study was to assess the effects of tebufelone on hepatocyte ultrastructure and hepatic cytochromes p450 (P450s) in the beagle dog after 2 weeks of oral administration at dose levels of 0, 5, 15, 50, and 100 mg/kg/day (N = 1/sex/dose level). Hepatic tissue was obtained at necropsy for histologic, ultrastructural, and biochemical evaluation. Hepatocellular hypertrophy was observed in only a single tebufelone-treated dog (50 mg/kg). Electron microscopic evaluation, however, revealed marked dose-dependent increases in smooth endoplasmic reticulum in all of the tebufelone treatment groups. Biochemical indicators suggested that tebufelone produced mixed effects on hepatic P450s. p-Nitroanisole O-demethylase and, to a greater extent, ethoxyresorufin O-deethylase activities were decreased with increasing tebufelone dose. The precise mechanism by which tebufelone decreased ethoxyresorufin O deethylase activity in dogs in unknown, but it was not by competitive inhibition, P450 inactivation, or reduced CYP1A expression. Tebufelone treatment increased NADPH-dependent cytochrome c reductase, total P450, and indicators of CYP2B11 (chloramphenicol covalent binding and immunochemically determined 2B11) and CYP3A12 (erythromycin N-demethylase, triacetyloleandomycin spectral complex formation, testosterone 6 beta-hydroxylase, and immunochemically determined 3A12). The largest increase in the 2B11 and 3A12 markers occurred in the 50 or 100 mg/kg treatment groups. The greatest increase in CYP2B11 markers produced by tebufelone treatment ranged from 2- to 3-fold, whereas the increase in CYP3A12 markers ranged from 5- to 10-fold. The changes in hepatic ultrastructure and increases in CYP2B11 and CYP3A12 markers produced by tebufelone in dogs are similar to that reported for phenobarbital. PMID- 8723732 TI - Caffeine induces cytochrome P4501A2: induction of CYP1A2 by tea in rats. AB - Previous reports indicated that treatment of rats with green tea or black tea extracts increased CYP1A2 activity, but such an induction was not observed with decaffeinated green tea in our preliminary study. Herein we report a comparative study on the induction of CYP1A2 with different tea preparations and caffeine as an inducer. When green tea (2%) or black tea (2%) was given to male Fischer 344 rats as the sole source of drinking fluid for 21 days, a 2.4- or 2.7-fold induction, respectively, of CYP1A2-dependent O-methoxyresorufin demethylase (MROD) activity in liver microsomes was observed. Treating rats with caffeine (0.04%) also resulted in an 1.9-fold increase in the MROD activity, but decaffeinated green tea (0.8%) did not cause such an induction. Rats treated with green tea (2%) or caffeine (0.055%) as the sole source of drinking fluid for 1, 3, and 7 days also showed comparable induction (from 1.7- to 2.1-fold) of the MROD activity. The induction was also shown by intragastric administration of caffeine (100 mg/kg). The induced MROD activity caused by consumption of green tea, black tea, and caffeine corresponded to the increase in liver microsomal CYP1A2 protein, as determined by immunoblot analysis. The concentrations of tea polyphenols and caffeine in plasma were also measured. Close correlation of the increase in the MROD activity was observed only with the plasma caffeine level (r = 0.736, n = 10, p = 0.015), not with the combined tea polyphenol level (r = 0.058, n = 6, p = 0.913). The present study establishes caffeine as an inducer of CYP1A2 and demonstrates that caffeine, not tea polyphenols, is the component in tea responsible for the induction of this enzyme. PMID- 8723733 TI - Effects of taurine supplementation in parenteral nutrition-associated hepatosteatosis and lidocaine metabolism. A study using isolated rat liver perfusion. AB - The effects of taurine in parenteral nutrition (PN)-related hepatic dysfunction are controversial. The aims of the present study were to evaluate the effects of taurine on hepatic function and on lidocaine metabolism, using two strengths of taurine (15 and 50 mg/dl) that are present in commercially available preparations. Animals (N > or = 4) were randomly assigned to one of the four 7 day treatment groups: chow-fed (CF); dextrose/amino acids (PN); dextrose/amino acids and taurine, 15 mg/dl (T15); dextrose/amino acids and taurine, 50 mg/dl (T50). Between 40-75% of the animals treated with PN developed steatosis. The origin of steatosis was zonal specific and dependent on taurine treatment. All livers in the CF group had a normal cellular architecture. Lidocaine metabolism was found to be impaired in groups PN, T15, and T50. This was indicated by a significant reduction in the intrinsic clearance values: 70%, 76%, and 85% in groups PN, T15, and T50, respectively (p < 0.05). Metabolites-to-drug ratios indicated that N-dealkylation, m-hydroxylation, and aryl methyl hydroxylation were significantly reduced in all treatment groups; the most pronounced effect was observed in the T50 group. These findings suggest that PN infusion results in impaired liver function, and the reduction of drug elimination rate is exacerbated by the addition of taurine. PMID- 8723734 TI - Suppression of constitutive cytochrome P450 gene expression by epidermal growth factor receptor ligands in cultured rat hepatocytes. AB - In rat hepatocytes cultured on Matrigel, incubation with epidermal growth factor (EGF) or transforming growth factor-alpha for 24 hr suppressed the constitutive expression of cytochrome P450 (CYP) 2C11 mRNA by 60-70%. The growth factors were maximally effective at concentrations of 10-30 ng/ml. These agents also suppressed the expression of CYP2C11 protein measured 48-72 hr after addition to the medium. Significant suppression of CYP2C11 mRNA was first seen 8 hr after EGF addition to the medium, was maximal by 16 hr, and persisted for at least 36 hr. The suppression of CYP2C11 mRNA by EGF was comparable in magnitude with that produced by interleukin (IL)-1, but greater than that by IL-6. The suppressive effects of EGF and IL-1 on CYP2C11 mRNA were additive, suggesting that the signaling pathways for suppression of CYP2C11 by EGF and IL-1 are different. PMID- 8723735 TI - Placental transfer of the hypolipidemic drug, clofibrate, induces CYP4A expression in 18.5-day fetal rats. AB - Rats at day 15.5 of gestation were dosed intraperitoneally with 300 mg.kg-1 of clofibrate for three consecutive days at 24-hr intervals and were culled 24 hr after the final injection. This regime produced maximal induction of the cytochrome P4504A (CYP4A) mRNAs in the maternal liver and kidney and in 18.5-day fetal tissues. The maternal hepatic and renal CYP4A mRNA levels had risen 12- and 2-fold, respectively, above the constitutive levels seen in untreated pregnant rats at an equivalent stage of gestation. Clofibrate was capable of traversing the placenta and modulating the fetal CYP4A mRNA expression as demonstrated by a 3-fold elevation in the mRNA levels in those fetuses explanted from drug-induced mothers, compared with those fetuses removed from untreated mothers. The CYP4A mRNAs were demonstrated in the fetal liver via dot-blot and Northern blot analyses. In addition, low levels of CYP4A mRNA expression were detected in the induced placenta via Northern blot analysis. Western blot analysis revealed that the CYP4A protein levels increased in the maternal liver and in the kidney and fetal livers after exposure to clofibrate. Peroxisome proliferation, a phenomenon associated with induction of CYP4A1 expression in rodents, was demonstrated in both maternal and fetal livers, with the use of light and electron microscopy. PMID- 8723736 TI - The aliphatic oxidation of salmeterol to alpha-hydroxysalmeterol in human liver microsomes is catalyzed by CYP3A. AB - Salmeterol xinafoate (Serevent) is a long-acting beta2-adrenoceptor agonist, used in the treatment of asthma, that has bronchodilator and anti-inflammatory action. Salmeterol is extensively metabolized by aliphatic oxidation in humans, with the major metabolite being alpha-hydroxysalmeterol. The aim of this investigation was to identify the specific cytochrome P450 (P450) isoform or isoforms involved in the formation of alpha-hydroxysalmeterol in human liver microsomes. [14C]Salmeterol was incubated with a pooled sample (N = 19) of human liver microsomes in the absence or presence of selective chemical inhibitors of the major human P450 isoforms. One microM ketoconazole, a selective inhibitor of CYP3A, substantially inhibited the metabolism of salmeterol to alpha hydroxysalmeterol. Disulfiram caused a small but consistent decrease in the amount of alpha-hydroxysalmeterol formed, possibly reflecting less than total selectivity for CYP2E1 under the conditions used. Other selective inhibitors had no significant effect on the metabolism of salmeterol. The rates of formation of alpha-hydroxysalmeterol in 10 individual liver microsomal samples showed an approximately 10-fold variation and were found to be highly correlated (r2 = 0.94; p < 0.001) with rates of metabolism of midazolam to 1'-hydroxymidazolam, a marker of CYP3A activity, in the same microsomal samples. No significant correlation was evident for the metabolism of salmeterol with levels of total P450 or other markers of human P450 activities in the same microsomal samples, thus indicating that the formation of alpha-hydroxysalmeterol is catalyzed predominantly by CYP3A. Insect cell microsomes that coexpressed human CYP3A and NADPH-P450 reductase were able to metabolize [14C]salmeterol to alpha hydroxysalmeterol, thus confirming the role of CYP3A in catalyzing this reaction. The therapeutic dose of salmeterol is very low, so it is unlikely that any clinically relevant interactions will be observed as a consequence of the coadministration of salmeterol and other pharmaceutical agents that are metabolized by CYP3A. PMID- 8723737 TI - Disposition of valpromide, valproic acid, and valnoctamide in the brain, liver, plasma, and urine of rats. AB - Valpromide (VPD) and valnoctamide (VCD) are amide derivatives of valproic acid (VPA), one of the major antiepileptic drugs (AEDs). In rodent models, both VPD and VCD are more potent as anticonvulsants than VPA. However, in humans, VPD served as a prodrug to VPA, whereas VCD acts as a drug on its own, which is not biotransformed to its corresponding acid--valnoctic acid (VCA). The present study investigates the pharmacokinetics (PKs of VPD and VCD in rats by monitoring the levels of these two amide isomers in the brain, liver, plasma, and urine of rats. The disposition of VPD and VCD was analyzed in a comparative manner with that of VPA. The following PK parameters were obtained for VPD and VCD, respectively: clearance, 6.1 and 3 ml/min/kg; volume of distribution (Vss), 0.63 and 0.58 liter/kg; half-life (t1/2), 42 and 94 min; and mean residence time (MRT), 102 and 196 min. The clearance of VCD in rats was half of that of VPD, and their Vss was similar. Therefore, VCD, t1/2, and MRT were twice as long as those of VPD.PK analysis of VPD and VCD liver and brain levels gave similar major PK parameters to those obtained from the plasma data. VPD underwent hepatic biotransformation to VPA, which persisted in the liver and brain for a longer period than VPD. The fraction metabolized of VPD to VPA was 42%. The brain was not found to be a metabolic site of the VPD-VPA biotransformation. Unlike VPD, VCD did not undergo amid-acid biotransformation to its corresponding acid, but was eliminated by biotransformation to unidentified metabolites. In contrast to VPD and VCD that distributed about evenly between the plasma, liver, and brain, VPA showed different disposition patterns in the plasma, liver, and brain. VCD and VPD distribute better into the brain than VPA, a fact that may contribute to their better anticonvulsant activity. PMID- 8723738 TI - Identification and characterization of limonene metabolites in patients with advanced cancer by liquid chromatography/mass spectrometry. AB - Limonene is a farnesyl transferase inhibitor that has shown antitumor properties. The drug had been given orally to cancer patients. Plasma and urine samples collected from the patients were examined by reversed-phase HPLC-atmospheric pressure chemical ionization and electrospray ionization MS. The drug underwent rapid conversion to hydroxylated and carboxylated derivatives. Characterization and structural elucidation of the metabolites were achieved by LC/MS and NMR. Five major metabolites were detected in the plasma extracts, namely limonene-1,2 diol, limonene-8,9-diol, perillic acid, an isomer of perillic acid, and dihydroperillic acid. Urinary metabolites comprised the glucuronides of the two isomers of perillic acid, dihydroperillic acid, limonene-8,9-diol, and a monohydroxylated limonene. PMID- 8723739 TI - Fate of 4"-epiacetylamino-4"-deoxyavermectin B1 in rats. AB - Distribution, excretion, and metabolism of 4"-epiacetylamino-4"-deoxyavermectin B1 (AAB1), a new avermectin, were determined in Sprague-Dawley VAF rats. The rats were dosed orally for 7 consecutive days at approximately 6 mg/kg body weight with [5-3H]AAB1 as a 1.2 mg/ml aqueous suspension containing 0.5% methyl cellulose. Rats were killed at approximately 7 hours and 1, 2, and 5 days after the last dose. The major route of excretion of drug residues was via feces, with less than 1% of the dose found in urine. The radioactive residue levels in tissues and blood followed the order GI > liver approximately equal to fat approximately equal to kidney > muscle > plasma approximately equal to red blood cells and were comparable in male and female rats. HPLC-radiochromatographic profiles revealed that 4"-epiamino-4"-deoxyavermectin B1a was the major metabolite in all tissue samples and plasma samples, and was usually the major residue at later time points. The results indicate that N-deacetylation of AAB1 was the primary route of metabolism in rats. A distinct feature of the metabolism was a sex difference in the extent of metabolism. When metabolite profiles of male and female rats killed at the same time were compared, less parent drug and more of the N-deacetylated metabolite were found in the female rats, indicating that the drug was metabolized more extensively in female rats than in male rats. The sex difference in the extent of metabolism was also demonstrated in vitro. PMID- 8723740 TI - p,p'-Dichlorodiphenyl sulfone metabolism and disposition in rats. AB - p, p'-Dichlorodiphenyl sulfone (DDS) is a lipophilic monomer used extensively in the synthesis of high temperature plastics. Studies of the fate of uniformly labeled [14C]DDS in the rat have established that it is readily absorbed from the gastrointestinal tract, distributed to all tissues examined, and concentrated in adipose tissue. After intravenous administration of 10 mg/kg and determination of the time course of DDS distribution, increasing accumulation of DDS in adipose was observed up to 24 hr, followed by slow elimination with a half-life of approximately 12 days. DDS equivalents in tissues were primarily (> 90%) parent compound, whereas excreted DDS equivalents were primarily (> 80%) present as metabolites. On repeat oral dosing at 10 mg/kg, levels of DDS in tissues seemed to reach steady state after approximately 2 weeks, at which time the concentrations in adipose reached 265 micrograms/g tissue. Hepatic cytochrome P450 (CYP) content, ethoxyresorufin O-deethylase activities, and levels of metabolites arising from phase I metabolism were doubled after repeat oral administration of DDS, but benzphetamine N-demethylase activity was unchanged. Thus, it seems that DDS induces CYP1A forms, but not CYP2B isozymes. DDS-derived radioactivity was excreted primarily in feces and to a lesser extent in urine as a phenolic metabolite and its glucuronide. The aglycone of this glucuronide was isolated and characterized by NMR and MS as 3-hydroxy-4,4'-dichlorodiphenyl sulfone. PMID- 8723741 TI - 17 beta-Estradiol metabolism by hamster hepatic microsomes. Implications for the catechol-O-methyl transferase-mediated detoxication of catechol estrogens. AB - We have shown that the metabolism of 17 beta-estradiol in hamster liver microsomes is concentration-dependent. At low (< 25 microM) concentrations of 17 beta-estriol, 16 alpha-hydroxylase activity predominated, and estriol was the major metabolite. At higher concentrations (25-75 microM), 16 alpha-hydroxylation and aromatic hydroxylation at C2 contributed equally to 17 beta-estradiol metabolism. Aromatic C4-hydroxylation was maximal at 75 microM of 17 beta estradiol and was always less than C2-hydroxylation. Dehydrogenation of the 17 beta-hydroxyl group to the ketone (estrone) was also observed, but both estrone and 2-hydroxyestrone were minor (approximately 3%) metabolites of 17 beta estradiol, only detectable at concentrations of 50 microM and above. Catechol-O methyl transferase (COMT) effectively converted both 2- and 4-hydroxyl-17 beta estradiol to their corresponding monomethoxy metabolites. Effective reducing conditions are required for COMT activity, because catechol estrogens are readily oxidized to their corresponding ortho-quinones, and ascorbic acid is routinely added to assays of COMT activity. Interestingly, although ascorbic acid (1 mM) increased the recovery of 2- and 4-hydroxy-17 beta-estradiol from microsomal incubations, it decreased the recovery of the methoxy metabolites (approximately 40%). Since the enediol function of ascorbate resembles that of a catechol group, ascorbate is a substrate for COMT and probably competes with the catechol estrogens for methylation. Because previous studies describing the ability of COMT to inhibit the covalent binding of electrophilic reactive metabolites of [4 (14)C]17 beta-estradiol to microsomal protein were performed in the presence of high (100 mM) Mg2+ concentrations, we also investigated the effects of Mg2+ on 17 beta-estradiol metabolism. Concentrations of Mg2+ > 10 mM inhibited the metabolism of 17 beta-estradiol, as evidenced by i) the increased recovery of substrate; ii) a decrease in the formation of estriol, estrone, and 2-, and 4 hydroxy-17 beta-estradiol; iii) a decrease in the recovery of water-soluble metabolites when incubations were performed in the presence of glutathione (GSH) to trap the reactive electrophilic metabolites; and iv) a decrease in the amount of reactive electrophilic metabolites bound to microsomal protein. GSH also decreased the covalent binding of electrophilic metabolites of [4-(14)C]17 beta estradiol to microsomal protein, with the concomitant formation of water-soluble metabolites. Thus, both COMT and GSH combine to limit the formation of electrophilic metabolites from 17 beta-estradiol. The relative importance of each of these pathways to the disposition of the catechol estrogens remains to be determined. PMID- 8723742 TI - 3-Hydroxymethylcholanthrene: metabolic formation from 3-methylcholanthrene and stereoselective metabolism by rat liver microsomes. AB - 3-Hydroxymethylcholanthrene (3-OHMC) was identified as one of the three initial hydroxylation products formed in the metabolism at the aliphatic carbons of 3 methylcholanthrene (3MC) by rat liver microsomes. The 3-OHMC formed in 3MC metabolism by liver microsomes prepared from untreated (control) rats, and rats treated with phenobarbital, 3MC, and polychlorinated biphenyls (Aroclor 1254) was determined by HPLC analysis and the effect of enzyme inducers on its formation was phenobarbital > polychlorinated biphenyls > 3MC > control. Incubation of 3 OHMC with rat liver microsomes produced the following identifiable products: 1 hydroxy-3-hydroxymethylcholanthrene (enriched in 1S-enantiomer, enantiomer excess 14-50%), 2-hydroxy-3-hydroxymethylcholanthrene (enriched in 2S-enantiomer, enantiomer excess 30-92%), 3-hydroxymethylcholanthrene-1-one, 3 hydroxymethylcholanthrene-2-one, 8-hydroxy-3-hydroxymethylcholanthrene, 3 hydroxymethylcholanthrene trans-9,10-dihydrodiol (enriched in 9R,10R-enantiomer, enantiomer excess 64-86%), 3-hydroxymethylcholanthrene trans-7,8-dihydrodiol (enriched in 7R,8R-enantiomer), and 3-hydroxymethylcholanthrene trans-11,12 dihydrodiol (enriched in 11R,12R-enantiomer). The enantiomer compositions were determined by circular dichroism spectral analysis and/or chiral stationary phase HPLC analysis. PMID- 8723743 TI - Characterization of the cytochrome P450 enzymes involved in the in vitro metabolism of dolasetron. Comparison with other indole-containing 5-HT3 antagonists. AB - Dolasetron mesilate [(2 alpha, 6 alpha, 8 alpha, 9a beta)-octahydro-3-oxo-2,6 methano-2H-quinolizin-8-yl-1H-indole-3-c arboxylate monomethane-sulfonate], is a 5-HT3 receptor antagonist, which is in development for the treatment of chemotherapy-induced emesis. The compound is rapidly reduced by carbonyl reductase to form its major pharmacologically active metabolite reduced dolasetron (red-dolasetron), which us further metabolized by cytochrome P450 (CYP450). Studies were conducted, using human liver microsomes, to characterize the CYP450 enzymes responsible for the in vitro metabolism of red-dolasetron. Red dolasetron underwent oxidation of the indole aromatic ring at positions 5, 6, and 7, and also N-oxidation. Enzyme-selective inhibition and correlation studies showed that hydroxylation of red-dolasetron was CYP2D6-dependent, and N-oxidation was conducted by CYP3A4. The rate of formation of 6-hydroxy red-dolasetron was significantly correlated with that of 5-hydroxy red-dolasetron, which further suggested that these metabolites were formed by the same CYP450 enzyme(s). Inhibition studies also demonstrated that 6-hydroxylation was, to a lesser extent, CYP3A4-dependent. This was confirmed by correlation experiments, wherein formation of this metabolite was significantly correlated with that of N-oxide formation, in quinidine-inhibited microsomes. Results were compared with those obtained with two other indole-containing 5-HT3 receptor antagonists: tropisetron and ondansetron. Tropisetron hydroxylation was CYP2D6-dependent, whereas that of ondansetron was both CYP2D6- and CYP2E1-dependent. Results were further confirmed, when compounds were incubated with microsomes containing recombinant human liver CYP2D6, CYP3A4, and CYP2E1. Red-dolasetron was a competitive inhibitor of CYP2D6, with an IC50 value of 70 microM, which is 2 orders of magnitude above maximum plasma concentrations found in humans. The implications of these in vitro results to the in vivo metabolism of these compounds in humans and their potential pharmacokinetic consequences is discussed. PMID- 8723744 TI - Formation of (R)-8-hydroxywarfarin in human liver microsomes. A new metabolic marker for the (S)-mephenytoin hydroxylase, P4502C19. AB - Kinetic studies demonstrate that two forms of human liver cytochrome P450 are responsible for the formation of (R)-8-hydroxywarfarin: a low-affinity enzyme (KM approximately 1.5 mM), previously identified as P4501A2; and a high-affinity enzyme (KM = 330 microM), now identified as P4502C19 on the basis of the following evidence. In crossover inhibition studies with P4501A2-depleted human liver microsomes between (R)-warfarin and (S)-mephenytoin, reciprocal competitive inhibition was observed. Apparent KM values for (S)-mephenytoin-4'-hydroxylation (52-67 microM) were similar to the determined Ki values (58-62 microM) for (S) mephenytoin inhibition of (R)-8-hydroxywarfarin formation. Similarly, the apparent KM for (R)-warfarin 8-hydroxylation in furafylline-pretreated microsomes (KM = 289-395 microM) was comparable with the Ki values (280-360 microM) for (R) warfarin inhibition of (S)-4'-hydroxymephenytoin formation. Inhibition studies with tranylcypromine, a known inhibitor of (S)-mephenytoin hydroxylase activity, and either substrate in three different microsomal preparations yielded nearly identical inhibitory constants: Ki = 8.7 +/- 1.6 microM for inhibition of (S)-4' hydroxymephenytoin formation and 8.8 +/- 2.5 microM for inhibition of (R)-8 hydroxywarfarin formation. In addition, fluconazole, a potent inhibitor of (R) warfarin 8-hydroxylation, Ki = 2 microM, was found to inhibit (S)-mephenytoin hydroxylation with an identical Ki (2 microM). Finally, a strong correlation between (S)-mephenytoin 4-hydroxylation and (R)-warfarin 8-hydroxylation activities in furafylline-pretreated microsomes was demonstrated in 14 human liver microsomal preparations (r2 = 0.97). PMID- 8723745 TI - Hepatic drug-metabolizing activities in rats after 14 days of oral administration of the human immunodeficiency virus-type 1 protease inhibitor ritonavir (ABT 538). PMID- 8723746 TI - The effect of dimethoate on bone marrow cell chromosomes of rats in subchronic four-generation experiments. AB - The effect of the insecticide dimethoate (O,O-dimethyl S-/N-methylcarbamoylmethyl phosphorodithioate) on bone marrow cells of four generations of rats was investigated. Male and female Wistar rats were treated for 6 weeks with 5 treatments per week via gavage at doses of 7.0, 9.33, and 14.0 mg/kg (1/ 100, 1/75, and 1/50 of the LD50). Chromosomes of 10 of the treated males were prepared. Other males (6-8 males per dose) were paired with females (11-16 females per dose). This represented the first (P) generation. During pregnancy and lactation females were treated with adequate doses of dimethoate. In F1 (70 males and 56 females), F2 (71 males and 62 females), and F3 generations (62 males and 44 females) treatment with dimethoate and the preparation of chromosomes (40 males per generation) were the same as those in the P generation. Following treatment with dimethoate in P and F1 generations, the number of numerical chromosome aberrations increased significantly. No significant changes in chromosome aberrations were found in F2 and F3 generations compared to the control groups. The frequency of numerical chromosome aberrations decreased considerably compared to the P generation at the dose of 14.0 mg/kg (1/50 of LD50). PMID- 8723747 TI - Effects of copper sulfate on black bindweed (Polygonum convolvulus L.). AB - Seedlings of black bindweed (Polygonum convolvulus L.) were transplanted to soil contaminated with copper sulfate at different dosages. These plants were followed until maturity under greenhouse conditions, and measures of mortality, growth, and reproduction were obtained from harvests 21, 34, and 105 days after the transplant. It was found that application of 125 mg Cu2+ kg-1 resulted in 1.7% lifetime mortality increasing to 100% at 500 mg kg-1. The mortality was dependent on exposure time. No further mortality was observed after 50 days. The population consisted of a relatively tolerant and a sensitive group. This was indicated by differences in lifetime of plants treated with 500 mg kg-1 and by a diversification in biomass of individuals dying from treatment with 315 mg kg-1. The plants surviving treatment had reduced biomass and seed production at dosages above 200 mg kg-1. PMID- 8723748 TI - Population level consequences of toxicological influences on individual growth and reproduction in Lumbricus rubellus (Lumbricidae, Oligochaeta). AB - The effects of increased environmental concentrations of copper on the population dynamics of Lumbricus rubellus are investigated. A size-structured matrix model is used to translate sublethal effects on individual growth and reproduction into their population dynamical consequences. Laboratory data on growth and reproduction under different, sublethal conditions of copper stress are used to parameterize the model. An estimate for the critical threshold concentration of copper (critical in a sense that the population growth rate at this concentration equals zero), obtained from the model analysis, agrees well with observations on field populations of L. rubellus. PMID- 8723749 TI - Toxicity of dimethoate to some soil animal species in different soil types. AB - Toxicity of dimethoate (insecticide) to an earthworm (Aporrectodea caliginosa tuberculata), a collembola (Folsomia candida), and an enchytraeid worm (Enchytraeus crypticus/variatus) was studied in three different soil types (artificial soil, clayey soil, and humus sandy soil). Parameters measured were survival and biomass change of the earthworms and survival and reproduction of the collembolas and enchytraeids. The degradation of dimethoate was analyzed too. Toxic effects were observed at the concentrations of some mg/kg dry soil. The biomass reduction of the earthworms occurred at lower concentrations than reduction in survival. The collembolas were more susceptible to dimethoate than the enchytraeids. High organic matter content of the soil reduced the toxic effects. The degradation of dimethoate was rather slow in the soils and under conditions tested. Half-life varied between 41 and 84 days. PMID- 8723750 TI - Effects of algae frozen at different temperatures on chronic assessment endpoints observed with Daphnia magna. AB - The Daphnia magna 21-day juvenile production test is not yet fully standardized because of many sources of variation. One is the diet provided to daphnids: the ration must be sufficient and the quality of algal cells must be optimal for achieving the required number of offspring defined by the new OECD guidelines. The experiments reported herein first examined the effects of Raphidocelis subcapitata after it had been maintained under four different conditions of storage (4, -20, -80, and -196 degrees C) on the survival, reproductive performance (over 21 days), and growth (ascertained by dry weight) of individually held D. magna for three generations. Under all of the four regimes tested, daphnids survived and reproduced in a manner which fulfilled the current OECD guidelines for a valid test, but the best results were obtained with fresh algae and algae frozen at -80 degrees C. Second, although D. magna has been widely used to determine toxicity of chemical substances, there are no reports in the literature that describe a rigorous study of the nutritional quality of the algae given to daphnids. Therefore, cell number, optical density, amount of organic carbon, and esterasic activity (assessed by intracellular breakdown of FDA to fluorescein) of algae that have been preserved at 4, -20, -80, and -196 degrees C were investigated. This part of the study indicated that freezing had no effect on cell numbers, in contrast to optical density, amount of organic carbon, and esterasic activity. First, it was found that esterasic activity was closely correlated to the reproductive performance of daphnids. It appears important, therefore, to consider the inclusion of this enzymatic activity as part of the routine quality control given to this microinvertebrate chronic procedure. PMID- 8723751 TI - Fate and biological effects of polymeric MDI (4,4'-diphenylmethane diisocyanate and homologs) in small artificial ponds. AB - The effects from a simulated accidental pollution event in a pond with polymeric MDI (4,4'-diphenylmethane diisocyanate and homologs) on different trophic levels of the aquatic ecosystem were investigated in small artificial ponds. Three 4.5 m3 volume ponds, interconnected with closable locks, were provided with natural lake sediment and ground water. Caged fish (rainbow trout, Oncorhynchus mykiss) were added to each pond, and the interconnecting locks were kept open to establish nearly identical physicochemical and biological conditions. At this stage, the ponds were isolated from one another and MDI was added at a dosage of 1 g/liter on top of the sediment of treated part of the first pond, 10 g/liter to the second pond, and 0 g/liter to the third pond (untreated control). Neither the applied monomer MDI nor its potential reaction product MDA (4,4' diphenylmethanediamine) was detected in water or accumulated by fish. The MDI polymerized to inert polyurea on the sediment of the test ponds. This polymerization formed carbon dioxide, released as bubbles which floated to the water surface. Some carbon dioxide was solubilized in water and reduced the water pH of about 9 by 2.0 units as an average in the high-dosed pond and 0.7 in the low-dosed pond. This reduction caused some other minor changes in the physicochemical characteristics of the pond water. Neither application rate caused any direct effect on the pelagic community (phytoplankton, zooplankton, fish, macrophytes) of the test ponds. Some minor indirect effects caused by the production of carbon dioxide were observed in phyto- and zooplankton community structures. Also, an increase of macrophyte growth was noted. Organisms living in the untreated part of the sediment (macrobenthos) were affected as a result of physical obstructions in this habitat. These populations, however, regained densities equivalent to the control after some weeks, except for Bivalvia which have too long of a generation time for the test period of this study. PMID- 8723752 TI - Sensitivity of Daphnia carinata sensu lato to the insect growth regulator, pyriproxyfen. AB - This paper reports the sensitivity of Daphnia carinata sensu lato to pyriproxyfen, a promising chironomid control agent. Neonates exposed to pyriproxyfen in acute toxicity trials produced 48-hr LC50 of 0.08 ppm, eight times the concentration known to control the emergence of nuisance chironomids. However, continuous exposure to pyriproxyfen (0.01 ppm) during a 3-brood, 14-day life cycle test, suppressed the growth of D.carinata and reduced reproduction by as much as 80%. PMID- 8723753 TI - In vitro effect of lead on Ca(2+)-ATPase in synaptic plasma membranes and microsomes of rat cerebral cortex and cerebellum. AB - Earlier investigations from this laboratory suggest that lead interferes with the calcium homeostasis of rat brain through modulation of an inositol polyphosphate second-messenger system. The present investigation was initiated to study the comparative effects of lead chloride and lead acetate on synaptosomal and microsomal Ca(2+)-ATPase of rat cerebral cortex and cerebellum in vitro. The synaptic plasma membranes and microsomes were prepared by using sucrose gradient (1.2-0.8 M). The assay of Ca(2+)-ATPase was done by hydrolysis of ATP and the liberated inorganic phosphate was estimated. Both lead chloride and lead acetate at micromolar concentrations significantly inhibited the Ca(2+)-ATPase of synaptic plasma membranes and microsomes of cerebral cortex and cerebellum in a concentration-dependent manner. The IC50 values of Ca(2+)-ATPase for both lead salts in synaptosomes were significantly lower (P < 0.05) than that of microsomes, indicating more sensitivity. Significantly (P < 0.05) lower IC50 values for both synaptosomal and microsomal Ca(2+)-ATPase were obtained for lead acetate than for lead chloride. The results suggest that lead acetate is more potent than lead chloride in inhibiting the Ca(2+)-ATPase. The microsomal Ca2+ uptake was also studied in cerebellum and cerebral cortex in the presence of different concentrations of both the lead salts. However, these lead salts in vitro did not reveal a significant (P < 0.05) change in the microsomal Ca2+ uptake of cerebellum and cerebral cortex. But earlier investigations indicated that in vitro lead (0.25-2 microM) inhibits inositol 1,4,5-triphosphate-mediated Ca2+ uptake and release in microsomes of rat cerebellum. PMID- 8723754 TI - Amitraz effects on foot-and-mouth disease virus in mammalian cells in vitro. AB - The toxicity of the acaricide amitraz and its effect on foot-and-mouth disease virus multiplication were evaluated in IB-RS-2 cells in vitro. A reduction of cell growth rate that was dependent on the dose and the length of treatment was observed in cells exposed to amitraz concentrations ranging from 20 to 50 micrograms/ml. Foot-and-mouth disease virus infectivity remained essentially unchanged in cells exposed to amitraz (20 micrograms/ml) 24 hr prior to virus infection or after the adsorption period. Viral RNA synthesis evaluated through [3H]uridine incorporation in cells treated for 24 hr prior to infection was not affected by amitraz. PMID- 8723755 TI - Changes in oxygen consumption and biochemical composition of the marine fouling dreissinid bivalve Mytilopsis sallei (Recluz) exposed to mercury. AB - The effect mercury (Hg) on tolerance, oxygen consumption, and body biochemical composition of a marine dreissinid bivalve Mytilopsis sallei was investigated. The 96-hr LC50 of Hg for M. sallei was found to be 0.2571 mg/liter and the safe concentration was 2.6 micrograms/liter. Exposure of M. sallei to Hg resulted in a significant decrease in oxygen consumption with increasing concentration of the metal. The effect of exposure time and concentration of Hg on body biochemical composition was also studied in M. sallei. In time-dependent experiments, carbohydrates were utilized, whereas both proteins and carbohydrates were consumed in concentration-dependent exposure. A decrease in the ratios of glycogen/protein and glycogen/lipid was observed in time-dependent experiments. In concentration-dependent experiments, there was also a decrease in glycogen/lipid ratio but the glycogen/protein ratio was almost constant at all exposure concentrations. The caloric concentration was also presented for both time and concentration-dependent exposure of Hg. However, the results of the present investigation indicate that this bivalve exhibited a differential preference in their utilization of biochemical constituents during time- and concentration-dependent stress of Hg. The decrease in oxygen consumption together with the utilization of glycogen and carbohydrates during Hg exposure suggests that these bivalves might shift to anaerobic metabolism in order to encounter the heavy metal stress in the environment. PMID- 8723756 TI - Fate and impact of pesticides applied to potato cultures: the Nicolet River basin. AB - The fate of cash-crop (potato) pesticides was monitored from the fields on which they were applied to the nearby streams. The investigation took place in the Nicolet River basin in the province of Quebec, Canada. The main pesticides under study were aldicarb, fenvalerate, metribuzin, and phorate. Aldicarb was never detected in any of the samples. The other pesticides were all detected in soils at low concentrations. Only fenvalerate and metribuzin were detected in tile drain. Metribuzin concentrations of up to 0.25 microgram/g were detected in the soil giving rise to a concentration of 1.3 micrograms/liter in tile drain and 47.1 micrograms/liter in surface runoff. Low concentrations of metribuzin up to 0.41 microgram/liter were detected in the nearby streams. The CREAMS model simulating pesticide movement in the fields overestimated metribuzin losses in the runoff at a concentration of 107 micrograms/liter. The subsurface EXPRES model using a PRZM time series adequately estimated a metribuzin field subsurface runoff concentration of 0.5 microgram/liter. According to the Canadian Water Quality Guideline for the protection of aquatic life, the concentrations of pesticides found in surface waters of this potato-growing region of Quebec do not have a potential to impact on the aquatic life in these systems. PMID- 8723757 TI - Innate immune function as a bioindicator of pollution stress in fish. AB - Immunotoxicological studies, based on processing of samples in the field and laboratory, were conducted on fish collected from a stream receiving point-source contaminants near its headwaters. Previous studies in this stream have revealed that cytochrome P4501A activity, liver somatic indices, macrophage aggregates, and parasitic liver lesions are significantly elevated in sunfish with the degree of impact decreasing with distance from the contaminant source. Fish collected from each sampling site were equally divided, One group was sacrificed in the field and the spleen and anterior kidney tissues were removed and placed in buffer on ice. The other group was kept in MS-222 for 2 hr and transported to the laboratory for processing. The spleen and anterior kidney from each fish were then prepared as a single cell suspension and shipped overnight to Mississippi State University. Cells were then evaluated for PMA-stimulated phagocyte oxidative burst and non-specific cytotoxic cell (NCC) activity against K562 tumor targets. Oxidative burst responses were dramatically suppressed in both groups at sampling sites near the headwaters but returned to reference levels further downstream. There were no differences between processing strategies at each station. NCC activities did not follow gradient-response patterns observed with phagocyte oxidative burst data and there were inconsistent differences between processing strategies at each site. These data indicate that simple immune function assays, such as phagocyte oxidative burst responses, can be used as a ancillary bioindicator in fish health monitoring and that immune function in these fish can be reliably assessed even if samples are not immediately processed. PMID- 8723758 TI - A comparison of the Ames assay and Mutatox in assessing the mutagenic potential of contaminated dredged sediment. AB - The ability of the Ames assay and of Mutatox to identify the genotoxic potential of dredged sediments was compared. The Ames assay has been used extensively in the testing of environmental contaminants, whereas Mutatox, a new bacterial bioluminescence test, has only recently been used for this purpose. Ten sediments with varying degrees of contamination were soxhlet extracted. Each of the 10 extracts was split with half remaining in a crude form and half cleaned using silica gel chromatography, resulting in 20 extract samples. Both the Ames assay (using Salmonella typhimurium strains TA98 and TA100) and Mutatox were conducted with and without S9 metabolic activation. When metabolically activated, TA98 and TA100 indicated a positive mutagenic response in 80 and 50%, respectively, of the sediment extracts. Without S9 activation, TA98 indicated a positive mutagenic response with half the extracts, whereas only 10% did so with TA100. Mutatox indicated a positive mutagenic response with S9 activation in 75% of the extracts and no mutagenic response in any of the sediment extracts without metabolic activation. In a side-by-side comparison of the Ames assay (TA98 with S9) and Mutatox, 80% of the sediment extracts had similar responses, both positive and negative. Fifty percent of the sediment extracts had similar responses when tested with TA100 and Mutatox in the presence of S9. Mutatox compared reasonably well with the Ames assay but was insensitive to the presence of direct-acting mutagens in the sediments tested. Although Mutatox demonstrates promise as a screening tool to assess sediment genotoxicity, the authors consider it appropriate to use the Ames assay as a confirmation for definitive investigations. PMID- 8723759 TI - Three forms of RPTP-beta are differentially expressed during gliogenesis in the developing rat brain and during glial cell differentiation in culture. AB - In situ hybridization and Northern analysis demonstrate that the three splicing variants of RPTP-beta have different spatial and temporal patterns of expression in the developing brain. The 9.5-kb and 6.4-kb transcripts, which encode transmembrane protein tyrosine phosphatases with different extracellular domains, are predominantly expressed in glial progenitors located in the subventricular zone (SVZ). The 8.4-kb transcript, which encodes a secreted chondroitin sulfate proteoglycan (phosphacan), is expressed at high levels by more mature glia that have migrated out of the SVZ. The three transcripts are also differentially expressed in glial cell cultures; O2A progenitors express high levels of the 9.5- and 8.4-kb transcript, whereas type 1 astrocyte progenitors predominantly express the 6.4-kb transcript. C6 gliomas also express high levels of the 6.4-kb transcript. Treating C6 cells with the differentiating agent dibutyryl cyclic-AMP (DBcAMP), induces a decrease in the 6.4-kb transcript and a corresponding increase in the 8.4-kb transcript. O2A cells grown in the presence of basic fibroblast growth factor (bFGF) and platelet-derived growth factor (PDGF) remain highly proliferative and undifferentiated, and continue to express high levels of RPTP-beta. However, when O2A cells are grown in conditions that induce oligodendrocyte differentiation, there is a marked decrease in the expression of the transmembrane forms of RPTP-beta, as determined by immunofluorescence. These results demonstrate that RPTP-beta expression is regulated during glial cell differentiation and suggest that the different forms of RPTP-beta perform distinct functions during brain development. PMID- 8723760 TI - Astrocyte-astrocytoma cell line interactions in culture. AB - Astrocytomas are the most common brain tumors arising in the CNS and account for 65% of all primary brain tumors. Astrocytes have been shown to have the highest predisposition to malignant transformation compared to any other CNS cell type. The majority of astrocytomas are histologically malignant neoplasm. Previous studies have shown that resident astrocytes are the first cell type to react to tumors and surround them. However, the role of these astrocytes in tumor formation and progression has not been determined. In the present study, we have co-cultured astrocytes with a permanent cell line S635c15 (derived from anaplastic astrocytoma) in order to understand the cellular interactions between astrocytes and astrocytoma cells. Our studies demonstrate that astrocytes in contact with the tumor cells become reactive and fibrous with an increase in glial fibrillary acidic protein (GFAP) immunoreactivity as early as 4 days in culture. By 8 days, astrocytes formed glial boundaries around the tumor cells which grew as round colonies. The astrocytic processes surrounding the tumor cells were also intensely GFAP positive. Since the behavior of these cells observed in culture is very similar to their interaction seen in vivo, this co culture system may serve as an in vitro model for astrocyte and astrocytoma cell line interaction and aid in our understanding of the molecular and cellular mechanisms during early stages of tumor formation and cell interactions. PMID- 8723761 TI - Recovery phase of acute experimental autoimmune encephalomyelitis in rats corresponds to development of endothelial cell unresponsiveness to interferon gamma activation. AB - Activation of the vascular endothelium is important in the development of inflammation. Activated endothelial cells (EC) express surface markers not expressed by quiescent EC. These surface markers augment adhesion reactions and leukocyte migration. We examined microvessel EC activation longitudinally in experimental autoimmune encephalomyelitis (EAE) in Lewis rats. CNS microvessels were isolated at 0, 3, 7, 12, 20, and 30 days post-inoculation (PI). Normal and CFA-injected rat microvessels do not express activation antigens (Ag). Increased expression of major histocompatibility complex (MHC) class II molecule and intercellular adhesion molecule-1 (ICAM-1) were detected on CNS microvessels from immunized rats at 7 days PI, prior to development of clinical signs, and at 12 days PI. Enhanced MHC class I molecule was seen only at 12 days. MHC class II molecule expression was focally expressed along microvessel fragments. By 20 days PI, EC did not exhibit increased levels of any of the markers tested. Perivascular cells (possibly pericytes), however, were found to express MHC class II molecule and ICAM-1 up to 30 days PI. During the recovery phase isolated CNS microvessels from MBP-immunized rats were unresponsive to IFN gamma-mediated endothelial activation. Unresponsiveness was independent of IFN gamma concentration. These results suggest that the endothelium is restored to functional quiescence during the recovery phase of acute EAE. PMID- 8723762 TI - Lipopolysaccharide stimulates differential expression of myristoylated protein kinase C substrates in murine microglia. AB - Microglia rapidly respond to lipoplysaccharide (LPS) by transformation from resting to active states and secretion of several neuro- and immuno-regulators including tumour necrosis factor alpha (TNF-alpha), interleukin 1 beta (IL-1 beta), and interleukin 6 (IL-6). With longer LPS treatment, microglia are converted to reactive or phagocytic states with characteristics similar to macrophages in inflammation and injury processes. We have investigated LPS mediated changes in two myristoylated substrates of protein kinase C (PKC): MARCKS (myristoylated alaninerich C kinase substrate) and MRP (MARCKS-related protein). Within 6 hours of addition, LPS induced a twofold increase in [3H]myristoylated and immunoreactive MARCKS protein and a sevenfold increase in MRP. The differential effect of LPS on expression of MRP vs. MARCKS was even more dramatic at the level of transcription: S1 nuclease protection assays revealed a 40-fold increase in MRP mRNA levels (maximum at 4-6 hours), whereas a threefold increase was observed for MARCKS. TNF alpha and colony-stimulating factor 1 (CSF 1), two cytokines which are induced by LPS, did not reproduce the observed effect of LPS on MARCKS and MRP gene transcription. CSF-1 also induced differential transcription of MRP, but of lower magnitude (threefold) and more sustained than by LPS. Accordingly, these two substrates for PKC are differentially up-regulated by LPS, apparently independent of TNF alpha or CSF-1. PMID- 8723763 TI - Trophic effect of cholera toxin B subunit in cultured cerebellar granule neurons: modulation of intracellular calcium by GM1 ganglioside. AB - Survival of cerebellar granule cells (CGC) in culture was significantly improved in the presence of cholera toxin B subunit (Ctx B), a ligand which binds to GM1 with specificity and high affinity. This trophic effect was linked to elevation of intracellular calcium ([Ca2+]i), and was additive to that of high K+. Survival was optimized when Ctx B was present for several days during the early culture period. 45Ca2+ and cell survival studies indicated the mechanism to involve enhanced influx of Ca2+ through L-type voltage-sensitive channels, since the trophic effect was blocked by antagonists specific for that channel type. Inhibitors of N-methyl-D-aspartate receptor/channels were without effect. During the early stage of culture Ctx B, together with 25 mM K+, caused [Ca2+]i to rise to 0.2-0.7 microM in a higher proportion of cells than 25 mM K+ alone. A significant change in the nature of GM1 modulation of Ca2+ flux occurred after 7 days in culture, at which time Ctx B ceased to elevate and instead reduced [Ca2+]i below the level attained with 25 mM K+. GM1 thus appears to serve as intrinsic inhibitor of one or more L-type Ca2+ channels during the first 7 days in vitro, and then as intrinsic activator of (possibly other) L-type channels after that period. This is the first demonstration of a modulatory role for GM1 ganglioside affecting Ca2+ homeostasis in cultured neurons of the CNS. PMID- 8723765 TI - Productive and non-productive binding of botulinum neurotoxin A to motor nerve endings are distinguished by its heavy chain. AB - Botulinum neurotoxin type A, a di-chain protein produced by Clostridium botulinum and responsible for botulism, blocks acetylcholine release from peripheral nerves by binding to the terminals, undergoing internalization and proteolyzing a protein essential for exocytosis. As butolinum neurotoxin is being used clinically for the treatment of dystonias and certain spasticities, deciphering the details of its specific targeting to cholinergic nerve endings has assumed great importance. Thus, interaction of butolinum neurotoxin type A with murine motor nerve terminals-a prime target in vivo-was investigated. Autoradiographic analysis revealed saturable, high-affinity interaction of radioiodinated toxin (0.4 nM) with two ecto-acceptor types, distinguished by an excess of the toxin's heavy chain which prevented only a fraction of this binding. Botulinum neurotoxin was also biotinylated through its free sulfhydryl groups, known not to be essential for neurotoxicity. Similar binding of this active derivative was, likewise, partially blocked by heavy chain, confirming the above results. This binding that is resistant to heavy chain equates to botulinum neurotoxin interacting with productive ecto-acceptors, leading to delivery to its cytosolic site of action, because heavy chain proved unable to antagonize toxin-induced neuromuscular paralysis. In contrast, it is deduced that botulinum neurotoxin bound to heavy chain-susceptible sites has a different fate, presumably due to trafficking via another route, and thus would be inefficient in causing neuroparalysis. PMID- 8723764 TI - Presence of calbindin D28K and GAD67 mRNAs in both orthotopic and ectopic Purkinje cells of staggerer mice suggests that staggerer acts after the onset of cytodifferentiation. AB - We used in situ hybridization to study the expression of GAD67 and calbindin D28K mRNAs in developing mouse cerebellar Purkinje cells. Both genes are expressed prenatally; calbindin D28K mRNAs can be detected in Purkinje cells of embryonic day (E) 15 mice, whereas GAD67 mRNAs first appear slightly later, in E16 mice. The stunted Purkinje cells of staggerer (sg/sg) mutant mice maintain calbindin D28K and GAD67 expression. Our data suggest that the sg/sg mutation does not interfere with the transcriptional activation of these two genes, and might therefore act after the induction of specific gene expression in developing Purkinje cells. PMID- 8723766 TI - Ultrastructural distribution of synaptophysin and synaptic vesicle recycling at the frog neuromuscular junction. AB - Synaptic vesicle recycling after intense acetylcholine (ACh) release was studied at the frog neuromuscular junction (NMJ) using the synaptic vesicle transmembrane protein synaptophysin as immunocytochemical marker of the synaptic vesicle membrane during the process of exo-endocytosis. ACh release in cutaneous pectoris nerve-muscle preparations was stimulated by three different means: K+, Cd2+ in Ca(2+)-free medium, and electrical stimulation in the presence of 4-aminopyridine (4-AP). Cd2+ stimulation produced synaptic vesicle depletion and nerve terminal swelling. Electrical stimulation in the presence of 4-AP produced a reduction in the number of synaptic vesicles, deep axolemmal infoldings, coated pits, and coated vesicles. K+ stimulation did not produce any observable ultrastructural changes. Synaptophysin was labeled using silver-intensified immunogold in dissociated muscle fibers. Unstimulated and K(+)-stimulated preparations showed synaptophysin immunolabeling associated only with synaptic vesicles. In contrast, in Cd(2+)-stimulated preparations, synaptophysin appeared along the axolemma, mainly at the active zones, and after electrical stimulation it appeared in both axolemmal infoldings and the remaining synaptic vesicles. The results show that when synaptic vesicle recycling is inhibited by Cd2+ in Ca(2+)-free medium, or when 4-AP is present during electrical stimulation, synaptic vesicle fusion is accompanied by translocation and incorporation of synaptic vesicle membrane proteins into the axolemma. However, during the latter condition, synaptic vesicles are recycled through coated vesicles arising from the axolemmal infoldings. Conversely, during physiological-like stimulation of ACh release by K+ the synaptic vesicles are rapidly recycled at the active zones, by a double and rapid process of exo-endocytosis, without collapse into the axolemma. PMID- 8723767 TI - Fatty acid binding protein is induced in neurons of the dorsal root ganglia after peripheral nerve injury. AB - Peripheral nerve trauma induces the expression of genes presumed to be involved in the process of nerve degeneration and repair. In the present study, an in vivo paradigm was employed to identify molecules which may have important roles in these processes. A cDNA library was constructed with RNA extracted from rat dorsal root ganglia (DRG) 3 days after a sciatic nerve crush. After differential hybridization to this library, several cDNAs were identified that encoded mRNAs that were upregulated in the DRG ipsilateral to the crush injury, as opposed to the contralateral or naive DRG. Approximately 0.15% of all the clones screened were found to be induced. This report presents the types of induced sequences identified and characterizes one of them, DA11. The 0.7 kb DA11 full length cDNA clone contains a 405 nucleotide open reading frame that encodes a putative protein of 15.2 kDa (135 amino acid residues) and is a member of the family of fatty acid binding proteins (FABP). The DA11 protein differs by one amino acid residue from the sequence of the C-FAPB protein and by eight residues from the sequence of mal1, proteins found in rat and mouse skin, respectively. Northern and Western blot analyses showed that the DA11 mRNA and protein were induced in the injured DRG. Furthermore, studies using antibodies generated against DA11 found that the DA11-like immunoreactivity was more pronounced in the nuclei of neurons located in the DRG ipsilateral to the sciatic cut than those located in the contralateral DRG. The induction of DA11 mRNA and protein in DRG neurons suggests, for the first time, the involvement of a neuronal FABP in the process of degeneration and repair in the nervous system. PMID- 8723768 TI - Inhibition of lipid peroxidation attenuates axotomy-induced apoptotic degeneration of facial motor neurons in neonatal rats. AB - The purpose of this study was to investigate the role of oxygen radical-induced lipid peroxidative mechanisms in trophic deprivation-induced apoptotic motor neuronal degeneration by testing the ability of the 21-aminosteroid lipid peroxidation inhibitor tirilazad mesylate (U-74006F) to attenuate the retrograde degeneration of facial motor neurons following axotomy in 14-day-old rat pups. On day 0, the right facial nerve of each rat was transected at its point of exit from the stylomastoid foramen. Pups were treated orally with either 10 or 30 mg/kg U-74006F or cyclodextrin vehicle 10 min before axotomy, and post-treated once a day from days 1 to 6, and then once every other day from days 8 to 21. The rats were sacrificed 3 weeks post-transection and the surviving motor neurons, identified through choline acetyltransferase immunocytochemistry, were counted in three regions (planes) in the facial nucleus. In vehicle-treated rats, 56.2% (region A), 50.6% (region B), and 57.4% (region C) of the motor neurons in the ipsilateral facial nucleus survived 21 days following facial nerve axotomy in comparison to the non-axotomized contralateral nucleus (P < 0.0001). Treatment with 10 mg/kg U-74006F significantly enhanced motor neuron survival in regions B and C to 72.8% (P < 0.01) and 66.7% (P < 0.02%), respectively. The 30 mg/kg dose level also increased survival rates to 64.2% (P < 0.02) and 67.9% (P < 0.01), respectively. A second experiment demonstrated that oral dosing with U-74006F (30 mg/kg), when limited to the first 5 days after axotomy, also significantly blunted retrograde degeneration measured at 21 days post-axotomy. The efficacy of the lipid peroxidation inhibitor U-74006F in protecting a portion of the facial motor neuron pool from post-axotomy degeneration suggests that lipid peroxidation may play a mechanistic role in trophic deprivation-induced apoptotic neuronal death. PMID- 8723770 TI - Toward an exhaustive sampling of the configurational spaces of the two forms of the peptide hormone guanylin. AB - The recently introduced Essential Dynamics sampling method is extended such that an exhaustive sampling of the available (backbone) configurational space can be achieved. From an initial Molecular Dynamics simulation an approximated definition of the essential subspace is obtained. This subspace is used to direct subsequent simulations by means of constraint forces. The method is applied to the peptide hormone guanylin, solvated in water, of which the structure was determined recently. The peptide exists in two forms and for both forms, an extensive sampling was produced. The sampling algorithm fills the available space (of the essential coordinates used in the procedure) at a rate that is approximately six to seven times larger than that for traditional Molecular Dynamics. The procedure does not cause any significant perturbation, which is indicated by the fact that free Molecular Dynamics simulations started at several places in the space defined by the Essential Dynamics sample that complete space. Moreover, analyses of the average free Molecular Dynamics step have shown that nowhere except close to the edge of the available space, there are regions where the system shows a drift in a particular direction. This result also shows that in principle, the essential subspace is a constant free energy surface, with well defined and steep borders, in which the system moves diffusively. In addition, a comparison between two independent essential dynamics sampling runs, of one form of the peptide, shows that the obtained essential subspaces are virtually identical. PMID- 8723769 TI - Flexible loop in the structure of S-adenosylmethionine synthetase crystallized in the tetragonal modification. AB - S-Adenosylmethionine synthetase (MAT, ATP:L-methionine S-adenosyltransferase, E.C.2.5.1.6.) plays a central metabolic role in all organisms. MAT catalyzes the two-step reaction which synthesizes S-adenosylmethionine (AdoMet), pyrophosphate (PPi) and orthophosphate (Pi) from ATP and L-methionine. AdoMet is the primary methyl group donor in biological systems. MAT from Escherichia coli was crystallized in the tetragonal modification with space group P4(3)2(1)2 using the same conditions as previously yielded crystals of the hexagonal system [Takusagawa, et al., (1996), J. Biol. Chem. 171, 136-147], except for the crystallization temperature. The structure has been determined by molecular replacement at 3.2 A resolution. The overall structure of the tetrameric MAT in the tetragonal modification is essentially the same as the structure found in the hexagonal modification. However there are two remarkable differences between the structures of two modifications. One is the contents in the active sites (holoform vs. apo-form), and the other is the conformation of the flexible loop over the active site (open vs. closed). These differences in the crystal structures are caused solely by the difference in crystallization temperatures (26 degrees C vs. 4 degrees C). We have interpreted the structural data obtained from the X-ray analyses in conjunction with the results of the mechanistic and sequencing studies in terms of possible dynamic motion of the flexible loop. When a substrate/product binds in the active site (hexagonal modification), the loop becomes disordered, apparently due to flexibility at the entrance of the active site as if it acts as a "mobile loop" during the catalytic reaction. On the other hand, when the temperature is decreased, the dynamic motion of the flexible loop may be reduced, and the loop residues enter the active site and close its entrance (tetragonal modification). Thus, the active site of the tetragonal modification is empty despite the crystals being grown in mother liquor containing a large concentration of phosphate (100 mM). There is no significant displacement of amino acid residues in the active site between the holo and apo forms, suggesting that the flexible loop plays an important role in determination of the contents in the active site. Since the functionally important amino acid residues in the active site are all conserved throughout various species, the structures of the active sites and the mechanism of the catalysis are probably essentially identical in the enzymes from a wide range of organisms. However, the substrate KM and Vmax values of MATs from various species are distributed over a wide range. The amino acid residues in the flexible loop regions are poorly conserved throughout various species. Therefore, the wide differences in catalysis rates of MATs from various speeches may be due to the differences in the composition of the flexible loop. PMID- 8723771 TI - Detailed description of an alpha helix-->pi bulge transition detected by molecular dynamics simulations of the p185c-erbB2 V659G transmembrane domain. AB - Molecular dynamics simulations of a 29-residue peptide including the transmembrane domain of the V659G mutant of the c-erbB2 protein demonstrate important dynamical behavior. Although the alpha helix is the structure commonly assumed for transmembrane hydrophobic segments, we found that hydrogen bond rearrangements can occur, giving rise to a structural deformation termed pi bulge stabilized by successive hydrogen bonds of pi helix type. A series of simulations enables us to give a detailed description, at the atomic level, of the alpha helix->pi bulge transition. The major consequence of this deformation covering one and a half turn of helix results in a noticeable shift around the helix axis of the C-Terminal residues relatively to those of the N-terminus. Such a deformation closely related to structural motifs described in the literature, induces a change in the distribution of the residues along the helix faces which could modulate the protein activity mediated by a dimerization process. PMID- 8723772 TI - Biopolymer-surfactant interaction: 4. Kinetics of binding of cetyltrimethyl ammonium bromide with gelatin, hemoglobin, beta-lactoglobulin and lysozyme. AB - The binding of CTAB with the proteins, gelatin, hemoglobin, beta-lactoglobulin and lysozyme follow first order kinetics and occurs either in two or three distinct stages. The number of stages depends on the overall configuration of the biopolymers. The denatured protein, gelatin has shown three-stage kinetics under all conditions, whereas the native proteins, hemoglobin, beta-lactoglobulin and lysozyme have exhibited two stage kinetics. Heat treated lysozyme in 8 mol dm-3 urea medium has also shown a two-stage kinetics. On the basis of non interacting binding sites on the proteins and independent sequential binding, the rates of reaction have been observed to increase with temperature and follow the trend k1 >> k2 > k3. The interaction of CTA+ with the proteins is both electrostatic and hydrophobic. Hemoglobin has shown maximum reaction rate whereas, beta lactoglobulin has shown a minimum. The activation parameters for the kinetic process have exhibited almost non-variant delta G++ and delta H++ < T delta S++. The formation of activation complex in the Eyring model is entropy controlled so also the overall kinetics. An isokinetic entropy-enthalpy compensation phenomenon has been observed for the respective kinetic stages. PMID- 8723773 TI - Simplified representation of proteins. AB - The conventional methods for characterizing the secondary structures of proteins based on hydrogen bonding patterns and phi,rho torsions are not fully specific in determining the spatial arrangements of various secondary structural elements. This fact motivates a search for an efficient description of the various secondary structures and their interactions. The successive identical repeating units of a polypeptide chain, namely the atoms of the peptide plane may be superposed using a method based on the mathematical quaternion. The superposition angle then characterizes different secondary structures. The distortions in protein alpha-helices such as kinks and bends are also precisely determined. The twist in beta-sheets is quantified and reverse turns are found to have characteristic variations. This new representation might pave the way for a better understanding of the final folding conformation of the polypeptide chain. PMID- 8723774 TI - An FTIR spectroscopic study of calf-thymus DNA complexation with Al(III) and Ga(III) cations. AB - The interaction of calf-thymus DNA with trivalent Al and Ga cations, in aqueous solution at pH = 6-7 with cation/DNA(P) (P = phosphate) molar ratios (r) 1/80, 1/40, 1/20, 1/10, 1/4 and 1/2 was characterized by Fourier Transform infrared (FTIR) difference spectroscopy. Spectroscopic results show the formation of several types of cation-DNA complexes. At low metal ion concentration (r = 1/80, 1/40), both cations bind mainly to the backbone PO2 group and the guanine N-7 site of the G-C base pairs (chelation). Evidence for cation chelate formation comes from major shifting and intensity increase of the phosphate antisymmetric stretch at 1222 cm-1 and the mainly guanine band at 1717 cm-1. The perturbations of A-T base pairs occur at high cation concentration with major helix destabilization. Evidence for cation binding to A-T bases comes from major spectral changes of the bands at 1663 and 1609 cm-1 related mainly to the thymine and adenine in-plane vibrations. A major reduction of the B-DNA structure occurs in favor of A-DNA upon trivalent cation coordination. PMID- 8723775 TI - The J-coupling restrained molecular mechanics (JrMM) protocol--an efficient alternative for deriving DNA endocyclic torsion angle constraints. Part I: Correlation of endocyclic torsion angles and vicinal torsion angle phi 1'2'. AB - An efficient alternative which makes use of the reliable 3J1'2' value to derive the endocyclic torsion angle constraints is proposed in this study. Based on the information embedded in the two plots, (i) the vicinal proton-proton J-couplings, 3J1'2', 3J1'2", 3J2'3', 3J2"3' and 3J3'4' against the pseudorotation phase angle, and (ii) 3J1'2", 3J2'3', 3J2"3' and 3J3'4' against 3J1'2'; using the calculated J couplings obtained for a range of sugar geometries of deoxyribose ring in nucleosides and nucleotides encountered along the pseudorotation itinerary [J. van Wijk, B.D. Huckriede, J.H. Ippel and C. Altona, Methods Enzymol. 211, 286-306 (1992)], it is suggested that the vicinal 3J1'2' possesses structural information other than the vicinal torsion angle phi 1'2'. This study is divided into two parts. In Part I, a correlation diagram between the endocyclic torsion angles nu(i) (i = 0,1,2,3,4) and the restrained vicinal torsion angle phi 1'2' is obtained through the use of the J-coupling restrained molecular mechanics (JrMM) protocol. The established phi 1'2'-nu(i) correlation shows nu(i) can be deduced from the reliable 3J1'2' value and it forms the basis for developing an alternative protocol to derive endocyclic torsion angle constraints. In Part II of this series, extensive testing demonstrating the validity of the JrMM protocol to derive nu(i) for defining the sugar geometry of solution DNA molecules is presented. PMID- 8723776 TI - The J-coupling restrained molecular mechanics (JrMM) protocol--an efficient alternative for deriving DNA endocyclic torsion angle constraints. Part II: Experimental application of the JrMM protocol. AB - The J-coupling restrained molecular mechanics (JrMM) protocol, which correlates deoxyribose endocyclic torsion angles and vicinal proton-proton torsion angle phi 1'2' in Part I of this study, is demonstrated to be a viable alternative to efficiently derive the endocyclic torsion angle constraints for the determination of the solution structures of DNA molecules. Extensive testing demonstrating the validity of the JrMM-derived torsion angle constraints in the restrained molecular dynamics and energy minimization structural refinement processes is performed theoretically using an energy-minimized B-DNA model and experimentally using a DNA hexamer d(CGTACG)2. The results show that only a 0.2 A difference exists between the RMSD values of the refined structures using the ideal and the JrMM-derived endocyclic torsion angle constraints. The JrMM-derived torsion angles are also determined to be in good agreement with the torsion angles derived through the use of the vicinal J-derived torsion angles. These results show that through the use of reliably measured J1'2' values and computer simulation method, the endocyclic torsion angle constraints can be derived reliably and efficiently. Thus the JrMM method serves as an alternative strategy to generate endocyclic torsion angle constraints for the determination of the solution structures of DNA molecules. PMID- 8723777 TI - Nonplanar DNA base pairs. AB - Three-dimensional structures of a representative set of more than 30 hydrogen bonded nucleic acids pairs have been studied by reliable ab initio quantum mechanical methods. We show that many hydrogen-bonded nucleic acid base pairs are intrinsically nonplanar, mainly due to the partial sp3 hybridization of nitrogen atoms of their amino groups and secondary electrostatic interactions. This finding extends the variability of intermolecular interactions of DNA bases in that i) flexibility of the base pairs is larger than has been assumed before, and ii) attractive proton-proton acceptor interactions oriented out of the base pair plane are allowed. For example, all four G-A mismatch base pairs are propeller twisted, and the energy preferences for the nonplanar structures range from less than 0.1 kcal/mol to 1.8 kcal/mol. We predict that nonplanarity of the amino group of guanine in the G(anti)...A(anti) pair of the ApG step of the d(CCAAGATTGG)2 crystal structure is an important stabilizing factor that improves the energy of this structure by almost 3 kcal/mol. Currently used empirical potentials are not accurate enough to properly cover the interactions associated with amino-group and base-pair nonplanarity. PMID- 8723778 TI - A molecular mechanics and dynamics study of alternate triple-helices involving the integrase-binding site of the HIV-1 virus and oligonucleotides having a 3'-3' internucleotide junction. AB - Triple helix formation by oligonucleotides can be extended beyond polypurine tracts with the help of specially designed linkers. In this paper we focus our attention on the integrase-binding site of the HIV-1 virus located on the U5 LTR end which contains two adjacent purine tracts on opposite strands. Two alternate triple helices with a 3'-3' junction in the third strand are considered: 5' GGTTTTp3'-3'pTGTGT-5' and 5'-GGAAAAp3'-3'pAGAGA-5' The structural plausibility of these triplexes is investigated using molecular mechanics and dynamics simulations, both in vacuo and in aqua. The non-isomorphism of the triplets in the GpT steps in the first sequence, gives rise to non canonical conformations in the torsion angles, hydration appears to be crucial for this triplex. Sugar puckers are predominantly South during in vacuo simulations while they turn East in aqua. In the simulation in aqua the triplexes are shrouded by an hydration shell, however, we have not been able to detect any permanent hydrogen bond bridge between DNA and water. The solvation of ions as well as their radial distribution, appear to be relatively well behaved despite the artifacts known to be generated by the simulation procedure. The experimental feasibility of these structures is discussed. PMID- 8723779 TI - DNA topological context affects access to eukaryotic DNA topoisomerase I. AB - We have analyzed the reactivity of a 217 base pair segment of the intrinsically curved Crithidia fasciculata kinetoplast DNA towards eukaryotic DNA topoisomerase I. The substrates were open [linear fragment and nicked circle] and closed minidomains [closed relaxed circle and circles with linking differences of -1 and -2]. We interpreted the results with the aid of a model that was used to predict the structures of the topoisomers. The modelling shows that the delta Lk(-1) form is unusually compact because of the curvature in the DNA. To determine the role of sequence-directed curvature in both the experimental and modeling studies, controls were examined in which the curved Crithidia sequence was replaced by an uncurved sequence obtained from the plasmid pBR322. Reactivity of the Crithidia DNA [as analyzed both by the cleavage and topoisomerization reactions] markedly varied among the DNA forms: (i) the hierarchy of overall reactivity observed is: linear fragment > nicked circular, closed circular [delta Lk(0)], interwound [delta Lk(-2)] > bent interwound [delta Lk(-1)]; (ii) the intensity of several cleavage positions differs among DNA forms. The results show that eukaryotic DNA topoisomerase I is very sensitive to the conformation of the substrates and that its reactivity is modulated by the variation of the compactness of the DNA molecule. The C. fasciculata sequence contains a highly curved segment that determines the conformation of the closed circle in a complex way. PMID- 8723780 TI - Characterization of polyion counterion interactions in cartilage by 23Na NMR relaxation. AB - Nuclear quadrupole relaxation is a sensitive measure of electrolyte environments. We used the relaxation of 23Na to probe mobile ion-matrix interactions and the electrostatic structure of the polyelectrolyte extracellular matrix of cartilage. Specifically, we measured spin-lattice and spin-spin relaxation times of 23Na in bovine nasal cartilage at 132 MHz under several conditions. Matrix fixed charge density was reduced by protonating anionic sites and by matrix digestion with trypsin and the relaxation times compared to controls. Under all conditions studied, measured longitudinal relaxation was monoexponential with values ranging from 16-32 msec. Transverse relaxation exhibited biexponential behavior in all cartilage samples with a fast component in the range of 2 to 5 ms and a slow component between 16 and 53 ms. Reduction in matrix fixed charge density in all cases led to a decrease in the relaxation rates. The results suggest a two-site model for Na+ ions in cartilage and a relaxation mechanism involving both polyion segmental motion and counterion diffusion. In the context of ion condensation theory, the implication of a two-site model is that the mean polyion-polyion spacing may be less than 0.7 nm. The mean polyion-counterion spacings were estimated by calculating correlation times and quadrupole coupling constants. These spacings were found to be 0.5-0.7 nm. PMID- 8723781 TI - The effect of nucleic acid geometry on counterion association. AB - The very high axial charge density of nucleic acids is a physical characteristic that substantially influences the thermodynamics of virtually all processes in which they are involved. This arises from long range electrostatic interacts between nucleic acids and the counter- and co-ions in solution so that salt concentration dramatically effects the activities of both reactants and products. A significant contributor to the resulting salt dependence for processes involving nucleic acids (e.g. ligand binding to a choice of nucleic acid substrates or a structural change), is the difference in arrangement of the sugar phosphate backbone of competing structures. This article reviews the results of a set of Grand Canonical Monte Carlo (GCMC) simulations that explores the effect of nucleic acid geometry, varied as a function of oligomer length and four-way junction branch length, on counterion association and therefore many nucleic acid processes. These GCMC simulations, which utilize a "primitive" model description of the nucleic acid, are complemented by a number of simulations which numerically solve the non-linear Poisson-Boltzmann equation utilizing detailed models for nucleic acids and proteins. Simulations of this kind are particularly useful for the study of systems that have been well characterized structurally, as well as thermodynamically. What is sought in the current article is insight into how an extremely general feature of DNA, namely the geometric arrangement of its phosphate charges surrounded by an exclusion surface, might play a role in determining nucleic acid processes. PMID- 8723782 TI - Umbilical cord blood: a novel alternative source of hematopoietic stem cells for bone marrow transplantation. PMID- 8723783 TI - Does CD34+ cell selection enrich malignant stem cells in B cell (and other) malignancies? PMID- 8723784 TI - Clinical challenges and cord blood transplantation: are we asking the right questions? AB - Cord blood offers an exciting alternative to traditional sources of stem cells for clinical transplantation, but its use will test traditional limits on minimal cell numbers required for engraftment as well as other aspects of the stem cell model of hematopoiesis. There is great interest in stem cell selection and ex vivo expansion techniques as they apply to cord blood. Although provocative, these approaches in their emphasis on the quantitative should not be allowed to obscure the important qualitative differences of cord blood with respect both to hematopoietic reconstitution per se and in the larger context to the unique immunologic status of that product. Whereas the momentum of cord blood transplantation may arise from the explosion in animal model-based stem cell biology, its direction must be determined by the clinical end points we week. PMID- 8723785 TI - Hematopoietic transplant potential of unrelated cord blood: critical issues. AB - To date, hematopoietic stem and progenitor cells from human umbilical cord blood (CB) have been employed in approximately 90 allogeneic (56 sibling and 34 unrelated) matched and mismatched transplantations worldwide with easy and successful restoration of hematopoiesis. Requests for stem cell preparations from CB will continue to increase. Thus, as a pilot study, the examination and standardization of unrelated cord blood-derived stem cell preparations and banking as well as their biologic characterization were initiated. Up to October 1995, a total of 574 samples [mean volume 79 +/- 26 ml, total nucleated cells (NC) 8.5 +/- 5 x 10(8), BFU-E 9.5 +/- 8.6 x 10(5), CFU-GM 5.7 +/- 6.3 x 10(5), CFU-GEMM 1.6 +/- 1.9 x 10(5)] from cord-derived or placental-derived residual blood have been defined by hematologic, immunologic, and microbiologic criteria. These CB samples were collected from the umbilical cord vein immediately after vaginal full-term delivery (n = 450) or cesarean section (n = 124) and stored frozen in liquid nitrogen. Seven percent of all samples collected could not be considered for potential transplants because of volumes < 40 ml. Only 5.0 ml of a CB sample is required for routine laboratory testing, consisting of HLA class I typing, HLA class II typing by sequence-specific oligonucleotide probes (PCR SSOP), ABO typing, sterility control, assessment of progenitor and stem cells by colony-forming and LTC-IC assays, and CD34+ status. To assess the potential problem of contaminating maternal cells, a PCR was performed on 7 representative samples. During the initial 6 months of the unrelated CB collection program, a median bacterial contamination rate of 18% (20% skin flora species, 80% perineal flora species) was encountered, which has since been reduced to < 1% through practical experience. With regard to viral infections, maternal sera was tested for HBsAg (0.6% positive), anti-HCV (0%), anti-HAV (IgG 18%, IgM 0%), anti-HIV-1 2 (0%), anti-EBV (IgG 98%, IgM 0%), anti-HTLVI-II (0%), anti-CMV (IgG 43%, IgM 0.4%), toxoplasmosis (46%) and syphilis (0%). In addition, all cord blood samples were tested by PCR for CMV infection. With regard to its clinical relevance, it is important that only 0.3% of all the samples were positive for CMV by this sensitive method. This may represent a critical advantage of CB grafts over bone marrow (BM) since, in contrast, > 40% of the unrelated BM donors have been identified to be positive for CMV. An additional advantage of CB is that since 20% of CB samples were collected from ethnic minorities, it appears possible to balance common HLA types and uncommon HLA types represented in this group. In summary, with the extensive practical experience of the obstetric collection team as well as the stem cell-processing laboratory, it appears feasible to obtain a 90% yield of unrelated CB-derived stem cell preparations for banking, which clearly should meet the medical and regulatory qualification criteria required for clinical transplantation. To test the feasibility of hematopoietic transplant potential of unrelated CB for adult patients, ex vivo expansion of CD34+-enriched stem/progenitor cell populations isolated from fresh or frozen CB was attempted in the presence of rh-IL-3, rh-IL-6, rh-EPO, rh-GM-CSF, and rh-SCF with or without fit 3. At varying time points (days 0, 2, 4, 7, 14, 21), the contents of these cultures were analyzed for the numbers of cells, CFC (BFU-E, CFU-GM, CFU GEMM), and LTC-IC. In this setting, the increase of cells was 200-fold, that of CFC 70-fold, and most importantly that of LTC-IC was 4.5-fold after 7 days in culture in the presence of flt3. In conclusion, LTC-IC derived from CB can be maintained and considerably expanded ex vivo from highly enriched CD34 + CB cell populations from fresh or frozen CB samples. PMID- 8723786 TI - The Milan Cord Blood Bank and the Italian Cord Blood Network. AB - We describe the activities of the Milan Cord Blood (CB) Bank and of the Italian Cord Blood Network. By October 31, 1995, 763 units were banked in Milan. Of these, 8 units were used to perform 4 related and 4 unrelated transplants in 5 children and 3 adult patients. Early cord clamping after delivery was found to be crucial to increase the volume of CB collected. This procedure does not seem to be detrimental to the newborn. Of various red cell sedimenting agents used to reduce the unit volume and concentrate progenitor cells, 3% gelatin seems to be associated with the best yields. After a preliminary experience of 2 years, the Italian Cord Blood Network (Gruppo Italiano Amplificazione Cellule Emopoietiche, GRACE) was founded in 1995. The initial activities of GRACE are aimed at the development at the national level of CB banking standard operative procedures in agreement with the draft issued by The North American Task Force for The Development of Standards for Hematopoietic Cell Transplantation. Moreover, a wet workshop has been organized to standardize colony-forming unit (CFU) evaluation. The main goal in Milan is the collection of 5000 CB units. Other issues of interest include CB volume reduction, hematopoietic progenitor purification, ex vivo expansion prior to transplantation, and experimental protocols for gene transfer, such as those related to the multidrug resistance (MDR) gene. PMID- 8723787 TI - Experience in autologous and allogeneic cord blood banking. AB - Transplantation and gene therapy offer the hope of treating thousands of individuals with various malignancies and genetic disorders. However, if this approach is to become a reality, there must be readily available sources of suitable HLA-A typed stem cells. Cord blood has been demonstrated to be a suitable source of hematopoietic stem cells that can be used in place of bone marrow stem cells for transplant. However, to implement the widespread clinical use of cord blood, it is necessary to establish both autologous and allogeneic cord blood banks from which stem cell specimens can be obtained. These cord blood banks must be large in size, reflect the ethnic diversity of the patient populations in need, and be financially and logistically feasible. At the Cord Blood Stem Cell Bank at the University of Arizona, we have addressed many of these issues and have taken the necessary steps to create both autologous and allogeneic cord blood banks. We have developed simple, efficient, and reproducible methods for cord blood collection that can be used by any physician without need for previous training. Further, we have developed methodologies to process the cord blood specimen so that samples can be stored in cryovials. This approach to cord blood banking allows for the storage of more than 10,000 independent cord blood specimens in a single liquid nitrogen freezer, reducing the costs and physical space requirements for such a facility. We also have developed the use of electronic integration of laboratory worksheets and controlled rate freezers that permit high laboratory throughput with reduced problems of reproducibility, data entry, and management errors. In terms of allogeneic cord blood banking, we have based our bank on several important assumptions regarding HLA haplotype frequency, ethnic diversity, and the need for less than perfect HLA matching between donor and recipient. These assumptions have allowed us to begin to create an allogeneic cord blood bank that can rapidly attain the size needed for clinical utility without exorbitant costs or time involved. Results of our efforts in autologous and allogeneic cord blood banking are presented (including factors that positively and negatively affect cord blood collection and banking). PMID- 8723788 TI - Issues in the development of a local cord blood bank. AB - An increasing number of transplants are being performed using stem cells harvested in placental and umbilical cord blood. Thus, interest in developing local cord blood banks is also increasing. This report discusses the issues and challenges that one may face in the development of a local cord blood bank. PMID- 8723789 TI - The applicability of cGMP to cord blood cell banking. AB - The Food and Drug Administration will regulate cord blood cells stringently as a distinct category of somatic cells. The FDA has declared that cord blood cell products are subject to licensure and will require a claimed exemption for an investigational new drug (IND) to undertake clinical trials. This regulatory approach will require both establishment and product licenses, requirements that will pose significant difficulties for regulated establishments. The manufacture of cord blood cell products will be divided between delivery rooms or attached laboratories and regional cord cell processing laboratories. It is possible that cooperative manufacturing arrangements, perhaps using a short supply agreement model, will be the most practical way for FDA to regulate cord blood cell product manufacture. Without regard to regulatory or business arrangements for providing cord blood cell products, it is clear that current good manufacturing practices (cGMP) will be required in both the delivery suite and the processing laboratory. The concepts of cGMP will be foreign to many medical practitioners, but embracing them will be crucial to comply with FDA regulatory expectations and will concomitantly ensure uniform product quality. PMID- 8723790 TI - Implementing a validation program in a cord blood bank. AB - The regulatory climate surrounding cord blood banking has been slowly evolving since the publication of the FDA notice, Applications of Current Statutory Authorities to Human Somatic Cell Therapy in the October 14, 1993, Federal Register. The pace of establishing regulatory policy for cord blood banking is about to quicken dramatically. Cord blood banking will most likely be regulated as a new blood product within the Office of Blood Research and Review in the Center for Biologics Evaluation and Research (CBER) and would then be subject to licensing preapproval inspections. Validation is a critical element of the approval process. Fundamental elements of structuring and implementing a scientifically sound validation program are discussed. PMID- 8723791 TI - Cord blood stem cells: how to get them and what to do with them. AB - This article reviews the means of obtaining cells from the available reservoirs of cord blood, intended as sources of immature hematopoietic stem cells that ultimately could be useful for transplantation, gene therapy, and research. Various issues must be considered when collecting umbilical cord blood regardless of the method employed. One must regard the basic fetal-placental physiology and hemodynamic characteristics prior to and at the time of procurement. Additional concerns exist with the mother, not only at the time of collection but also prenatally, including informed consent, health history, and psychosocial issues. Collection methods may be characterized as either ex utero or in utero, employing either open or closed collections methods. Each of these variables presents limitations and offers specific advantages over the others. Once collected, the cells must be appropriately tested, processed, and prepared for cryopreservation if not used immediately, using good manufacturing practices and acceptable standards of operation. An ideal collection method has yet to be defined that fulfills the need for reliability, reproducibility, and ease of use. PMID- 8723792 TI - Collection and cryopreservation of cord blood for the treatment of hematopoietic disorders: the obstetrician's overview. AB - Bone marrow transplantation provides successful treatment for many diseases of the immune and hematopoietic systems. The main therapeutic ingredient in this procedure is stem cells collected from the bone marrow. Recently, it has been demonstrated that stem cells from human umbilical cord blood can serve as an alternative to bone marrow transplantation in children. Although cord blood transplantation in adults has not yet been attempted, it appears that there are enough stem cells present in cord blood for successful engraftment in adults. Obstetric health care providers should be aware that many familial conditions are treatable by cord blood stem cell transplantation in children. Obstetric health care providers caring for patients with familial disorders should consider counseling such patients regarding the collection and storage of cord blood for potential future uses in autologous or allogeneic transplantation. PMID- 8723793 TI - Methods of isolation and cryopreservation of stem cells from cord blood. AB - Successful reconstitution of the hematopoietic system by umbilical cord blood (UCB) stem cells has sparked interest in large-scale banking of UCB. This would only be feasible if UCB samples could be frozen in small volumes. Early efforts to fractionate UCB produced significant losses of stem and progenitor cells. Several groups have proposed techniques to concentrate the stem/progenitor cell fraction of UCB, with good recovery. These are described, together with the principles of cryopreservation of stem cells. It seems feasible to store UCB samples on a large scale, although the different fractionation methods need to be compared, and the optimal, most efficient technique must be determined. PMID- 8723794 TI - Immune functions of cord blood cells before and after transplantation. AB - There have been numerous reports of decreased acute and chronic graft-versus-host disease in patients receiving HLA-matched or HLA-disparate umbilical cord transplants. It has been proposed that this may be due to the unique properties of the neonatal immune system, which permit the development of tolerance to alloantigens. This review discusses experimental evidence contrasting the immune functions of cells derived from cord blood and those from peripheral blood. PMID- 8723795 TI - Circulating progenitor cells in human ontogenesis: response to growth factors and replating potential. AB - We have measured the number of progenitor cells circulating in fetal (17-32 weeks of gestation), perinatal (36 weeks of gestation), and adult (30-50 years old) blood. The progenitor cells at each ontogenetic stage were also characterized in terms both of the minimal combinations of growth factors they required to form maximal numbers of colonies in vitro and of their self-replication potential, as measured by the number of secondary and tertiary progenitor cells each could generate. The number of progenitor cells circulating in fetal and perinatal blood can be measured by directly plating the unfractionated blood. In this assay, fetal blood contains half the number of progenitor cells detected in perinatal blood (18.0 +/- 16.4 versus 40.88 +/- 0.63, p < 0.01), and the number of progenitor cells in adult blood is below the level of detection of the assay (< 1/8 microliter of blood). To compare the number of progenitor cells in all three stages of human development, progenitor cell counts were performed on blood mononuclear cells enriched by density separation. In this case, the light density cell fractions from fetal and neonatal blood contained the same number of progenitor cells (300/10(5) cells), numbers that were 10-fold higher than those observed with adult blood (30/10(5) cells). Circulating fetal-neonatal erythroid and multipotential progenitor cells were found to differ from their adult counterparts in terms of their response to growth factors and their self-renewal ability. In fact, the number of cytokines required to observe maximal colony formation increased with the ontogenetic stage of the cells. No differences were found in the frequency of primary colonies containing progenitor cells or in the mean number of secondary progenitor cells per primary colony in cultures of fetal, neonatal, or adult blood. Differences between the three ontogenetic stages, however, were found with respect to the number of sequential replatings that were possible. In fact, although both secondary granulocyte-macrophage (GM) and mixed-cell colonies derived from fetal cells gave rise to tertiary colonies, only perinatal secondary mixed-cell colonies grew in tertiary cultures, and no growth was observed in tertiary cultures of adult cells. These results suggest that the greater amplification of progenitor cells observed in liquid culture of fetal/neonatal versus adult blood is due both to a higher proliferative capacity of neonatal progenitor cells (up to two replatings versus one) and to a higher frequency in these samples of mixed-cell colony-forming cells (CFC) (37.7 +/- 7.3 versus 2.0 +/- 0.7/10(5) light density cells, respectively). Because of the high numbers of progenitor cells circulating in the fetus, as well as their high proliferative capacity, it is predicted that if blood could be harvested directly in utero, fetal blood would be as good a source of stem cells for transplantation as perinatal placental/cord blood. Circulating fetal stem cells would, therefore, represent an ideal target for gene therapy and in utero autologous transplantation. PMID- 8723797 TI - CD34+ progenitor cell selection: clinical transplantation, tumor cell purging, gene therapy, ex vivo expansion, and cord blood processing. AB - A unique avidin-biotin immunoadsorption system (CellPro CEPRATE SC) has been developed for the rapid clinical scale purification of CD34+ cells from bone marrow, mobilized peripheral blood (PBSC), or cord blood. This system has been used to treat more than 2500 patients worldwide in a variety of clinical studies. In the autologous transplantation setting, selection for CD34+ cells results in significant reduction (2- > 5 logs) of contaminating tumor cells in the enriched fractions, as documented using sensitive immunocytochemistry staining techniques or quantitative PCR analyses. In the allogeneic bone marrow or PBSC transplant setting, using the CEPRATE SC system as a method for T cell depletion has shown a 3-log reduction in overall T cell content, with more rapid hematopoietic recovery after PBSC reinfusion as compared with bone marrow. CD34+ enrichment is required for efficient ex vivo expansion of progenitor cells as well as for gene therapy applications involving the genetic modification of stem cells. PMID- 8723796 TI - Inhibitory effects of interleukin 12 on retroviral gene transduction into CD34 cord blood myeloid progenitors mediated by induction of tumor necrosis factor alpha. AB - Interleukin 12 (IL-12), a heterodimeric cytokine with potent biologic activity, was evaluated for effects on retroviral-mediated gene transduction into human myeloid progenitor cells in vitro. Cord blood CD34 cells were prestimulated with Steel factor (SLF), IL-3, GM-CSF, and erythropoietin (Epo) in the presence and absence of 5-80 ng/ml IL-12 for 40 hr in suspension culture prior to gene transduction using viral supernatant collected from a packaging cell line containing the pLNL6 vector encoding Neo sequences. After gene transduction, cells were assayed for colony formation stimulated by Epo, GM-CSF, IL-3, and SLF, and gene transduction efficiency was determined by the percentage of G418 resistant (R) colonies and confirmed by PCR analysis. IL-12 dose-dependently inhibited retroviral-mediated gene transduction into human cord blood CD34 granulocyte-macrophage (CFU-GM) and erythroid (BFU-E) progenitors. These suppressive effects could be neutralized by incubation of IL-12 with polyclonal antihuman IL-12. IL-12 had no inhibitory effects directly on colony formation. To understand the possible mechanisms for this suppression, ELISA assays were used to detect the release of interferon (IFN)-gamma and tumor necrosis factor (TNF) alpha, which could potentially have been induced by IL-12 from CD34 cells. TNF alpha protein release was significantly increased in CD34 cells incubated with IL 12. No detectable levels of IFN-gamma were noted. Anti-TNF-alpha, but not anti IFN-gamma, blocked the inhibitory effects of IL-12 on gene transduction. Moreover, TNF-alpha, but not IFN-gamma, suppressed gene transfer to the same degree as IL-12. No change of amphotropic receptor mRNA expression was noted by Northern blot analysis in cells treated with or without IL-12. The results suggest that the suppressive effects of IL-12 on retroviral gene transduction are, at least in part, mediated by IL-12 induction of the release of TNF-alpha. PMID- 8723798 TI - Quo vadis? Fetal tissue transplantation. AB - The epidemiology and biologic characteristics of fetal tissue harvested from elective and spontaneous abortions are reviewed. The use of fetal bone marrow obtained from second trimester lost pregnancies is discussed. Allogeneic fetal tissue transplantation carried out in utero is reviewed. Data on intrauterine transplantation of human fetal bone marrow obtained from second trimester lost pregnancies into baboon fetuses are presented. The viability of this tissue, its clonogenic efficiency, engraftment, use in the future, and banking are discussed. PMID- 8723799 TI - Fetal tissue banking: standards and regulatory issues. AB - There has been a long-standing interest in collection and study of fetal tissues for the purpose of understanding normal ontogeny, as well as aberrant processes in development. As the unique features and capabilities of fetal tissues have become elucidated, it is evident that fetal tissues could potentially be used to ameliorate adult degenerative diseases through transplantation. Indeed, there has been significant work surrounding the transplantation of fetal-derived hematopoietic stem cells, islet cells, and central nervous system cells. Many involved with collection and transplantation of fetal tissues seem poised on the edge of the federal regulatory playing field. This article discusses the history of regulations related to cellular and tissue-derived products and current regulatory issues facing cell and tissue banks in the United States from the tissue bank perspective and focuses on these issues as they relate to the use of cells derived from fetal tissue. PMID- 8723800 TI - In utero transplantation of fetal liver stem cells into human fetuses. AB - Based on our experience in the field of fetal liver transplantation (FLT) that we have developed since 1976, we initiated, in 1988, in utero FLT into human fetuses, taking advantage of the immunologic tolerance in young fetuses. The transplants have involved fetuses suffering from various diseases at 12-28 weeks postfertilization, with 2 of the 6 cases eventually resulting in abortion. With the 4 other fetuses, a favorable outcome was observed. Three children are more than 4 years old, and they are alive and well, with evidence of engraftment, reconstitution of immunity, and partial correction of beta-thalassemia. In the fourth case, the fetus is alive and well and birth is expected soon. In utero transplantation of stem cells is a therapy with remarkable advantages: (a) tolerance induction due to immune immaturity of the host, (b) lack of graft versus-host disease (GVHD) due to immaturity of the donor, (c) ideal isolation of the fetus in the maternal uterus, and (d) optimal environment for donor fetal cell development in the vicinity of host fetal cells and growth factors. PMID- 8723801 TI - The impact of early obstetric discharge on newborn health care. AB - The impetus for shorter hospital stay of mother and newborn infant after delivery is based on economic constraints and parental preference. Earlier published studies did not demonstrate any increase in morbidity rate with shorter stay, but these studies were limited by methodologic flaws and biases that limited the validity and generalizability of the conclusions. More recent studies showed that readmission rates increased with shorter stay and that the severity of illness of readmitted infants may have increased. In addition, the interpretation of current newborn screening tests may not be applicable when performed prior to early discharge. In light of recent changes in neonatal hospital length of stay, a careful review and update of current guidelines and practices for newborn care are required. PMID- 8723802 TI - Management of painful procedures in the newborn. AB - Management of painful procedures in the neonate is less than optimal. Although evidence exists to support the infant's capacity to feel pain at birth, health professionals do not prescribe or administer adequate analgesia. This is largely because of the lack of published data on the safety and efficacy of pharmacologic interventions and lack of sufficient understanding of the benefits of nonpharmacologic measures. In this review, recent research on the safety and efficacy of pharmacologic and nonpharmacologic interventions for managing painful procedures in the neonate is summarized. PMID- 8723803 TI - Current knowledge of Kangaroo Mother Intervention. AB - Kangaroo Mother Intervention (KMI) started in 1978 in Colombia as a way of dealing with overcrowding and scarcity of resources in hospitals caring for low birth weight infants. Currently the intervention comprises three components: kangaroo position (skin-to-skin contact), kangaroo nutrition (exclusive or nearly exclusive breast-feeding), and kangaroo discharge policies (early discharge in kangaroo position regardless of weight or gestational age). Different authors have adopted and adapted diverse components of the KMI to suit the particular needs of their parents. We discuss different modalities of kangaroo care reported in developed and in developing countries and also describe in some detail the components of the whole KMI program. In addition, results from a systematic review of kangaroo-related papers published in English between 1991 and 1995 are provided, together with a summary of current knowledge (evidence-based) and research needs. PMID- 8723804 TI - Use of surfactant in pulmonary disorders in full-term infants. AB - Pulmonary surfactant is routinely used for treatment of premature babies with respiratory distress syndrome, and its use in full-term infants with respiratory failure is now under extensive clinical investigation. Several clinical reports have shown that intratracheal instillation of large doses of bovine and porcine lung-derived surfactant preparations has the potential to improve blood oxygenation and ventilation efficiency in full-term neonates with respiratory distress syndrome, meconium aspiration syndrome, pneumonia, pulmonary hemorrhage, and lung hypoplasia. These findings are suggestive of secondary surfactant deficiency or surfactant dysfunction in the pathogenesis of respiratory failure in full-term infants. Further clinical investigation with multicenter, randomized, controlled clinical trials is warranted to verify new indications of surfactant application for these diseases. PMID- 8723805 TI - The role of nitric oxide in the treatment of neonatal pulmonary hypertension. AB - Nitric oxide production appears to be decreased in infants with persistent pulmonary hypertension (PPHN). Inhaled nitric oxide may improve oxygenation by two mechanisms: increased pulmonary blood flow and improved ventilation-perfusion matching. Nitric oxide inhalation has been tested in newborns with PPHN, congenital heart diseases, and bronchopulmonary dysplasia. We present a review of the articles concerning inhaled nitric oxide for infants with PPHN. Overall, 59% of the neonates had an initial improvement in oxygenation in response to nitric oxide inhalation. A sustained response was observed in 60% of the infants. Patients with extrapulmonary shunting, clear chest radiographs, and adequate lung volume seem to have a better response, whereas patients with congenital diaphragmatic hernia, severe sepsis, and alveolar capillary dysplasia are more likely to fail. To define the benefit-risk ratio, six prospective randomized trials are currently in progress. PMID- 8723806 TI - Pediatric renovascular hypertension. AB - Renovascular disease is a frequent cause of severe hypertension in children and may result in significant morbidity or mortality. Most children presenting with renovascular hypertension have few if any symptoms, but devastating neurologic injury and congestive heart failure are still too often observed. Several new radiographic techniques have been used to detect renovascular lesions, but none has yet demonstrated consistently superior results when compared with intra arterial digital subtraction angiography. Renal venous renin sampling, duplex ultrasonography, and captopril-enhanced renal scintigraphy may be useful diagnostic adjuncts. Therapeutic objectives include cure of hypertension and restoration or preservation of renal function. At many institutions, percutaneous transluminal angioplasty has become the treatment of choice for patients with renal transplant artery stenosis and discrete, nonostial stenoses caused by fibromuscular dysplasia. More extensive lesions generally respond well to surgical correction. Chronic pharmacologic therapy is reserved for patients who do not respond to angioplasty or in cases in which the location or extent of involvement of the renal arterial system precludes surgical revascularization. Nephrectomy is usually reserved for kidneys that have minuscule function of irreparable vascular anomalies. An individualized approach to therapy is increasingly emphasized. PMID- 8723807 TI - An update on peritoneal dialysis and hemodialysis in the pediatric population. AB - Automated forms of peritoneal dialysis are used by the majority of children receiving chronic renal replacement therapy. Hemodialysis is performed in a smaller but substantial population of patients. Studies of peritoneal membrane solute transport conducted with exchange volumes standardized by body surface area reveal similar characteristics in children and adults. Target urea clearance and creatinine clearance values that define "adequate" dialysis for the pediatric peritoneal dialysis and hemodialysis populations have not yet been determined. Limited data are available to characterize the ideal dialysis access, whereas the clinical complications of infection, growth retardation, and patient mortality receive a great deal of attention. PMID- 8723808 TI - Primary nephrotic syndrome of childhood. AB - Primary nephrotic syndrome encompasses the spectrum of minimal change disease, focal segmental glomerulosclerosis, and related disorders. It is a common cause of morbidity and, more rarely, of mortality in children. Recent progress has permitted us to better understand the mechanism of proteinuria and how it causes the other findings of nephrosis. Developments related to the pathogenesis of nephrosis and why some patients develop focal segmental glomerulosclerosis are less striking, but recent studies have indicated useful directions for subsequent work in these areas. Finally, application of newer concepts to treatment has provided both a better prognosis for these diseases and decreased morbidity from treatment. PMID- 8723809 TI - Etiology, pathophysiology, diagnosis, and management of chronic renal failure in children. AB - There are a large number of causes of chronic renal disease in children. This review updates recent findings on conditions that arise in childhood that lead to chronic kidney disease in adulthood (eg, polycystic kidney disease, reflux nephropathy) or cause chronic kidney disease in childhood. Recent studies addressing the etiology and genetics of chronic kidney disease, as well as its pathophysiology, prognosis, and treatment, are discussed. PMID- 8723810 TI - A 9-year-old girl with vomiting, abdominal pain, and pancreatic and renal cysts. PMID- 8723811 TI - An historical perspective on the roots of managed care. AB - To most people, managed care and managed competition are terms of relatively recent origin, but the concepts are far from new. The use of these terms and their principal predecessors, prepaid group practice and medical care foundations, are symbolic of the underlying problems of the health care systems identified in the United States. With the passage of time, the individuals who first spoke loudly and well on the subject have come and gone, but the central theme of what they had to say was that major changes in the organization, financing, and delivery of health care are absolutely essential if high-quality preventive, diagnostic, and therapeutic care are to be available to the population at an acceptable cost. This paper reviews the early years of recognition of the need for change and the sporadic start of the changes. The presentation leaves out details on the recent growth of managed care and the political turmoil caused by the high level of medical costs, the inadequate documentation of the quality of care, and the large numbers of people who have no insurance or at best inadequate insurance. PMID- 8723812 TI - Managed care, primary care, and quality for children. AB - In an effort to provide medical care that is both more effective and less costly, the new variants of managed care organizations have instituted a variety of incentives and administrative controls that impact on the types and quantity of care provided to patients. Evidence suggests that the early forms of managed care, namely prepaid group practices, showed particular promise in improving the primary care delivered to children, ie, care that is accessible, person-focused in the long term, comprehensive, coordinated, and oriented toward achieving better outcomes. However, recent evidence concerning the quality of care delivered to children in the newer variants of managed care is mixed and scant; the newer organizational forms may not facilitate and may even have a negative impact on the attainment of primary care. Managed care can have a positive effect on first contact care, because it contractually defines a primary care provider and reduces use of the emergency room as a source of care. It may, however, have mixed effects on other aspects of access and use, depending on the plan's particular characteristics. Longitudinality is threatened by the disruption of prior relationships with out-of-plan providers and by the instability of both enrollees and providers in managed care plans. Children's benefits in managed care arrangements tend to include more preventive services, but access to specialty services has generally been found to be more restrictive. Coordination of care is not inherent to managed care, and many plans are no more likely to foster communication than are traditional indemnity plans. Evidence for the superior clinical quality afforded to children by new variants of managed care is lacking. Because managed care arrangements are proliferating rapidly, better studies are needed to prove or refute the contention that managed care has a significant positive effect on quality of care. PMID- 8723813 TI - Understanding different models of health maintenance organizations. AB - More than 50 million Americans belong to a health maintenance organization (HMO), and enrollment in all types of managed care plans continues to grow. To survive and thrive in the evolving health care system, independent and group practitioners find it essential to understand managed care. This article describes the key features and main characteristics of the most prevalent models of HMOs, as well as the role of physicians in each. It discusses the theoretical and practical benefits and drawbacks of HMOs, including financial and organizational relationships with physicians and other providers, variation in consumer choice, provider incentives, cost effectiveness, and management oversight. PMID- 8723814 TI - A capitation primer for pediatricians. AB - This paper discusses the basic issues involved in capitation contracting for pediatricians. We provide a blueprint for analyzing capitation arrangements. We begin by reviewing the basic concepts of capitation. We then discuss contract features, underwriting and pricing risks, administration, risk assessment and management, and finally quality of care considerations. PMID- 8723815 TI - A 10-year experience in pediatric after-hours telecommunications. AB - A rapidly growing area within the field of telephone medicine is the use of centralized telephone triage systems to help in the after-hours coverage of pediatric practices. This paper describes a 10-year experience with a pediatric telecommunications program within the 302,000-member Health Centers Division of Harvard Pilgrim Health Care. Telephone volume averages 3,000 calls per month, and over 175,000 telephone calls have been received since the program began. This article highlights the linkage to the primary care physician, the enhancement of service by the automated medical record system, and the application of the telecommunications program to resident teaching. The literature review focuses on the rationale for structured telecommunications programs, including improved quality of care, reduction of medicolegal risk, and the potential for reimbursement of services. PMID- 8723816 TI - Special perinatal services in a large health maintenance organization. AB - The Kaiser Permanente (KP) Medical Care Program, the largest and oldest health maintenance organization in the United States, is generally considered a model and template for other health maintenance organizations and managed health care programs. Nationally, KP has almost 7 million members-about 5 million in California, 2.4 million of whom are in the KP Northern California Region (KPNCR). About 500,000 of those are children; physicians in the program see 32,000 prenatal patients and deliver more than 28,000 babies annually. KPNCR is the largest KP region; 26% of northern California's population are subscribers. This review addresses special perinatal programs in the KPNCR and emphasizes perinatal screening and genetic services. PMID- 8723817 TI - Child abuse, sudden infant death syndrome, and psychosocial development. AB - A review of the literature on child abuse continues to emphasize the importance of careful attention to physical findings. Children who are allegedly sexually abused very often have no abnormal physical findings, yet they may be subjected to repeat examinations in an attempt to document possible physical effects of the abuse. Information is reviewed about the potential psychologic impact of these repeated assessments on young children. Controversy regarding the etiology of sudden infant death syndrome persists, and risk factors are reviewed. Changes in recommendations for infant sleep position by the American Academy of Pediatrics are not universally implemented. The importance of death scene investigations in cases of sudden unexplained infant death is emphasized. A review of the current research on infant colic does not provide many new insights, and the etiology remains controversial. The primary care pediatrician has an important role in providing advice and counseling, albeit on an empiric basis. PMID- 8723818 TI - Human papilloma virus infection of the vulva. AB - The relationship between human papilloma virus (HPV) and cervical cancer is in the process of being defined. This potential opportunity to understand a human oncogenic virus has drawn significant attention to HPV. While cervical cancer is a potentially fatal outcome of HPV infection, genital warts are the most common manifestation of genital HPV infection. In addition to knowledge of etiology, natural history, and therapeutic options, patients and providers need to consider emotional impact to successfully manage the care of patients with this common infection. This article summarizes current knowledge of recent advances of HPV infection of the vulva. PMID- 8723819 TI - Herpesvirus infections of the vulva. PMID- 8723820 TI - Candida vulvovaginitis. PMID- 8723821 TI - Vulvar disease in the pediatric population. PMID- 8723822 TI - Dermatitis of the vulva. PMID- 8723823 TI - Lichen sclerosus. PMID- 8723824 TI - Erosive vulvar lichen planus and desquamative vaginitis. PMID- 8723825 TI - Vulvar squamous cell carcinoma. AB - Squamous cell carcinoma of the vulva is a rare disease, mainly seen in elderly women. Risk factors are advanced age, an immunocompromised status, longstanding vulvar dystrophy, VIN, a history of vulvar human papillomavirus infection, and a history of cervical cancer. Vulvar cancer should be considered as a skin tumor and detection is possible in an early stage. However, because of patients' and doctors' delay, one in three vulvar cancers is not treated before an advanced stage. The tumor metastasizes mainly lymphatogenic. Spread starts in the inguinal lymph nodes. In the middle of this century, standard treatment, consisting of an en bloc dissection of the vulva and inguinal lymph nodes has been developed and applied. As a result, considerably improved survival rates were achieved: up to 90% 5-year survival rates for patients without lymph node metastases. However, complication rates were high. In recent years, a more individualized approach has replaced standard treatment. Surgical treatment now depends on the localization, size and extent of the tumor, and is followed or preceded by radiotherapy in selected cases. The role of chemotherapy in advanced disease is currently being studied in several referral centers. The most important success in the treatment of vulvar cancer in recent years is the maintenance of high survival rates despite considerably less extensive surgical treatment, resulting in lower complications rates. An important challenge for the near future will be the improvement of the management of advanced disease. However, an even more difficult issue may be the prevention of such large lesions. The reduction of treatment delays requires a considerable effort in education of both health care workers and the general public. PMID- 8723826 TI - Paget's disease. PMID- 8723827 TI - Vulvar melanoma. AB - Vulvar melanomas are seen in about 3% of all melanomas. The percentage reaches 10% of all malignant tumors of the vulva. The mean age of the patients is approximately 55 years. The first symptoms noticed by the patients are bleeding, pruritus, or a mass in the groin. Vulvar melanomas are mostly located at the labia minora. The differential diagnosis comprises different pigmented lesions, such as seborrheic keratosis, nevocellular nevus, lentigo, or hyperpigmentations of different origin. There is no typical histopathological pattern, but mostly melanomas of this site are of the mucosal-lentiginous type. As treatment modalities, most investigators propose a complete excision for melanomas thinner than 1 mm, and for thicker melanomas a wide excision. Recent investigations have shown that hemivulvectomies or vulvectomies do not show a better survival. The question of a prophylactic ipsilateral lymph node dissection is not answered yet. The survival rates depend on the tumor thickness and/or the infiltration level. The importance of an early diagnosis should be stressed in vulvar melanomas, especially through an accurate inspection of the vulva at the time of a genital examination. In addition, vulvar self examinations are an excellent possibility for early diagnosis. This allows a definite treatment and vulvar conservation. PMID- 8723828 TI - Treatment of vulvar dermatoses. PMID- 8723829 TI - Nerve regeneration through autogenous vein grafts: an SEM evaluation. AB - The role of autogenous vein grafts in nerve regeneration was studied in a rat model. Three groups of animals underwent autogenous vein grafting for ischiatic nerve regeneration, and were sacrificed 30, 60, and 90 days after surgery. Samples obtained were processed for S.E.M. study. Nerve regeneration consisted of two stages: the first (up to 60 days) was characterized by the presence of active endothelial cells; the second (after 60 days) by cell disappearance, with the venous wall acting as a protective conduit throughout both steps. Complete qualitative morphologic and functional studies of neural regeneration are presented, confirming previous investigations. PMID- 8723830 TI - Microsurgical reconstruction during the early puerperium. AB - A transverse rectus abdominis musculocutaneous free flap was electively performed for reconstructing the cutaneous cover in a patient during the early puerperium (12 hr after delivery). This flap was chosen because of the natural expansion achieved by pregnancy, and because it would cover the entire area to be reconstructed. During surgery, considerable vasodilation and continuous bleeding were encountered, as well as severe spasm of the arteries and veins both in the flap pedicle and at the recipient site. This spasm eased when local papaverine was used, and blood flow was interrupted but recurred as soon as the artery or vein had contact with the patient's blood. The administration of systemic papaverine caused dilation of flap and recipient vessels, but also of uterine vessels, with severe hemorrhage. Under such circumstances, the systemic use of papaverine was interrupted and systemic oxytocin was administered, with subsequent decrease in tissue perfusion and necrosis of the transplanted flap. PMID- 8723831 TI - The use of artificial dermis on the donor defect of the free forearm flap. AB - Microsurgical grafting of a free flap is an established technique of surgical reconstruction. The forearm free flap is frequently used following removal of oral and/or pharyngeal tumors. However, simple split-thickness skin grafting on full-thickness defects of the skin on the flexor side of the forearm results in deformations, pigmentation, and the adhesion of the graft to the tendons. The authors attempted to minimize such problems by repairing the flap donor site with a collagen sponge used as an artificial dermis. This artificial dermis was used to reconstruct postoperative defects in the skin of 5 patients. Split-thickness skin measuring 0.25 to 0.30 mm was grafted onto the defect. The donor site skin was successfully repaired in 2 to 3 weeks without adverse effects, without the development of depressed deformations, and with only slight pigmentation. The artificial dermis formed a matrix on the donor wound bed, with good cosmetic results. PMID- 8723832 TI - The effect of combined treatment with dextran 40 and acetylsalicylic acid on patency in severely traumatized small veins and arteries: an experimental study in the rabbit. AB - The effects of pharmacologic intervention on the fates of severely traumatized small veins and arteries have been studied in a rabbit model. Controls were given bolus doses of saline and a group treated with a combination of dextran 40 and acetylsalicylic acid starting prior to traumatization and continuing until postoperative day 5. Relative to controls, bleeding times in the treated group were significantly lengthened in arteries but not in veins, and venous patency significantly improved throughout the interval ending 2 weeks postoperatively. Arterial patency was at first highly improved but by 2 weeks, occlusion was virtually 100 percent. Since some of the occlusions took place more than a week after traumatization, the effects of antithrombotic agents on patency may need to be evaluated over considerably longer time periods than has previously been the rule. PMID- 8723833 TI - The management of unsalvageable free flaps. AB - The loss of vascular flow in the early postoperative period will generally lead to free flap failure. When attempts at flap salvage are unsuccessful, conservative management with delayed flap debridement may be indicated. Seven unsalvageable free flaps were managed with observation and flap debridement 4 to 14 days following loss of vascular signals. At the time of debridement, six of the seven wounds had viable granulation tissue and were successfully closed with skin grafts. The seventh patient had loss of vascular flow to the free flap within 12 hr of surgery and, at the time of delayed debridement, had no evidence of granulation ingrowth. Local revascularization of flaps is known to occur and offers an explanation for these findings. Delayed debridement of unsalvageable free flaps is indicated for noncritical wounds, and may obviate the need for a second free-tissue transfer to obtain wound closure. PMID- 8723834 TI - Microsurgical composite tissue transplantation at difficult recipient sites facilitated by preliminary installation of vein grafts as arteriovenous loops. AB - In microvascular surgery when local recipient vessels are inadequate, vein grafting is required. There are several potential inherent disadvantages of immediate vein grafting, including the development of graft thrombosis or leakage, an increased opportunity for technical errors, and an increased number of anastomoses in series. All of these may contribute to a higher failure rate for composite-tissue transplantation requiring vein grafts. The authors hypothesized that in cases where vein grafting is obviously required, the creation of a temporary, looped, arteriovenous fistula (AVF) would reduce the morbidity of vein grafting, by allowing the detection of thrombosis or technical errors predisposing to thrombosis prior to free-tissue transplantation. Since delaying the division of an AVF for 5 or more days may allow time for healing of the endothelium at the AVF anastomotic site, the hypothesis was that composite tissue transplantation whose vein grafts were installed as an AVF divided in a delayed manner, might have better patency than those in which vein grafts were installed at the time of reconstruction. This study reviews the results of 16 patients (8 females, 8 males) who underwent 17 microvascular reconstructions using AVFs. Patient courses and outcomes were compared between those undergoing immediate (8 patients) and delayed (7 patients) AVF construction, division, and free-tissue transplantation. There was a low patency rate for AVFs which were divided in a delayed fashion (2 of 7 patients, 29 percent), compared with those which were immediately divided (10 of 10, 100 percent). These results suggest that, in spite of a strong theoretical basis for delayed division of the AVF, delayed harvesting of the AVF is empirically associated with a higher thrombosis rate (p = 0.0048, Fishers exact test). PMID- 8723835 TI - Comparison of vascular effects of calcitonin gene-related peptide and lidocaine on human veins. AB - Arterial and venous constriction during microsurgery are among the leading causes of flap failure. So far, pharmacologic tools have been proposed only for arterial constriction, and local anesthetics are commonly used to counteract the vascular spasm. The purpose of this study was to investigate the vascular effects of two known arterial vasodilators on human veins, in order to evaluate the possibility of their potential use during microsurgery. In vitro experiments were performed on 31 greater saphenous vein (SV) rings obtained from 20 patients and on 12 cephalic vein (CV) rings from 7 patients. The rings were mounted in organ baths and their isometric contractile activity was measured. Experiments were conducted by cumulatively adding calcitonin gene-related peptide (CGRP) or lidocaine to the organ baths. The endothelium was mechanically removed in 18 SV rings and in 6 CV rings. Results demonstrated that in vitro CGRP produced vasodilation only; no constrictive effects were seen. Lidocaine produced a biphasic response with contraction at low concentrations (1.5 x 10(-5) M, 1.5 x 10(-3) M) and release of the maximal contraction at higher concentrations, similar to that observed in arteries. Removal of the endothelium did not significantly affect contractile activity in either CGRP- or lidocaine-treated vessels. The data supported the conclusion that CGRP could be used to relieve venous constriction, and it is suggested that further studies on the clinical use of CGRP in microsurgery is warranted. PMID- 8723836 TI - Cross-over replantation after bilateral traumatic lower-leg amputation: a case report with a six-year follow-up. AB - A suicidal 66-year-old female patient was run over by a train. The lower extremities were amputated at different levels. On the right side, there was a complete amputation in the distal third of the lower leg. Proximal to the amputation site, there was an extensive soft-tissue and bone defect. On the left side, there was a crush injury of the tarsus and mid-tarsal bones. The left lower leg showed only minimal injuries. An ipsilateral (anatomic) replantation was not possible. In order to save one lower extremity, a cross-over (contralateral) replantation of the right foot to the left lower leg was carried out. After a follow-up of 6 years, the patient is able to walk well with a prosthesis on the right side, and the right foot replanted to the left lower leg. From a psychological point of view, it seemed better for the patient to preserve one extremity, even with a cross-over replanted foot. PMID- 8723837 TI - The bipedicled iliac crest flap. AB - The use of the iliac crest flap based on the deep circumflex iliac artery and vein is well established in orofacial reconstruction. In cases where the deep circumflex iliac artery and vein are of inadequate size, or when large composite flaps are required, vascular augmentation can be achieved using the iliac branches of the iliolumbar artery and vein as an additional pedicle. Routine inclusion of this additional pedicle in iliac crest flaps creates extra safety with a low effort. PMID- 8723838 TI - Young neurons from the adult subependymal zone proliferate and migrate along an astrocyte, extracellular matrix-rich pathway. AB - The subependymal zone (SEZ) of the lateral ventricle of adult rodents has long been known to be mitotically active. There has been increased interest in the SEZ, since it has been demonstrated that neuroepithelial stem cells residing there generate neurons in addition to glia in vitro. In the present study, we have examined parasagittal sections of the adult mouse brain using immunocytochemistry for extracellular matrix (ECM) molecules (tenascin and chondroitin sulfate-containing proteoglycans), glial fibrillary acidic protein (GFAP, a cytoskeletal protein prominently expressed by immature and reactive astrocytes), RC-2 (a radial glial and immature astrocyte cytoskeletal marker), TuJ1 (a class III beta-tubulin isoform expressed solely by postmitotic and adult neurons), nestin (a cytoskeletal protein associated with stem cells), neuron specific enolase, and bromodeoxyuridine (BrdU, which is taken up by dividing cells). Our results demonstrate that a population of young neurons reside within an ECM-rich, GFAP-positive astrocyte pathway from the rostral SEZ all the way into the olfactory bulb. Furthermore, BrdU labeling studies indicate that there is a high level of cell division along the entire length of this path, and double labeling studies indicate that neurons committed to a neuronal lineage (i.e., TuJ1+) take up BrdU (suggesting they are in the DNA synthesis phase of the cell cycle), again along the entire length of the SEZ "migratory pathway." Thus, the SEZ appears to retain the ability to produce neurons and glia throughout the life of the animal, functioning as a type of "brain marrow." The implications of these findings are discussed in relation to the role that such a glial/ ECM-rich boundary (as seen in the embryonic cortical subplate and other developing areas) may play in: confining the migratory populations and maintaining them in a persistent state of immaturity; facilitating their migration to the olfactory bulb, where they are incorporated into established adult circuitries; and potentially altering SEZ cell cycle dynamics that eventually lead to cell death. PMID- 8723839 TI - 5-Hydroxytryptamine2A receptors on cultured rat Schwann cells. AB - Intracellular calcium responses of cultured rat Schwann cells to 5 hydroxytryptamine (5-HT) were examined using the calcium indicator dye fluo-3. Consistent changes in [Ca2+]i were observed with bath application of 5-HT and the basis of these responses was characterized. Application of 5-HT elicited a transient increase in intracellular calcium in a subpopulation of cultured Schwann cells. In many responding cells, the response recurred at approximately regular intervals following the initial transient. In some cases, these oscillations lasted for hours following removal of 5-HT from the bath. The increase in intracellular calcium evoked by 5-HT still occurred in the absence of extracellular calcium, suggesting that 5-HT induces calcium release from intracellular stores. Consistent with this hypothesis, the response to 5-HT was prevented by depletion of inositol trisphosphate-sensitive intracellular calcium stores with thapsigargin. Bath application of caffeine, known to activate Ca2+ release from ryanodine receptor-mediated stores, did not elicit an increase in [Ca2+]i. These results also suggested that 5-HT acted by stimulating a member of the 5-HT2 receptor family since this family employs inositol trisphosphate as a second messenger. In agreement with this interpretation, it was found that the 5 HT-induced intracellular calcium transients could be reversibly blocked by both ketanserin and spiperone, suggesting that the transients are mediated by 5-HT2A receptors. Additional support for this conclusion was obtained by immunocytochemistry using an anti-idiotypic antibody that recognizes a subset of 5-HT receptors. PMID- 8723840 TI - Schwann cell heparan sulfate proteoglycans play a critical role in glial growth factor/neuregulin signaling. AB - Glial growth factors are proteins encoded by the neuregulin gene and are thought to signal via receptor tyrosine kinases. Many neuregulin gene products bind heparin, and we hypothesize that affinity for heparin may implicate cell surface heparan sulfate proteoglycans (HeSPGs) as co-receptors for the soluble neuregulin gene product, recombinant human glial growth factor 2 (rhGGF2). Using primary rat Schwann cell cultures, we show that exogenous heparin and heparan sulfate block rhGGF2-induced phosphorylation of putative neuregulin receptors, and block subsequent DNA synthesis; other glycosaminoglycans show no such effect. Inhibition of Schwann cell HeSPG biosynthesis by administration of beta-xyloside also blocks responsiveness to rhGGF2. In cell-free binding assays, rhGGF2 binds heparin and heparan sulfate with high affinity, while suramin and suramin-like molecules block this binding. These suramin-like molecules reversibly block Schwann cell responsiveness to rhGGF2 with a rank order of potency identical to that in the cell-free binding assay. Thus we demonstrate high affinity and specificity in the interaction of rhGGF2 with heparin-like molecules, and show that three distinct perturbations of this interaction on Schwann cells (exogenous heparin/ heparan sulfate treatment, inhibition of HeSPG biosynthesis, and treatment with suramin-like molecules) result in a loss of responsiveness to rhGGF2. These results support a model in which HeSPGs are critical components that modulate extracellular rhGGF2 signaling interactions with appropriate receptor tyrosine kinases. PMID- 8723841 TI - In vivo characterization of endogenous proliferating cells in adult rat subcortical white matter. AB - Proliferating cells in adult rat subcortical white matter were characterized in vivo using stereotactic injections of a replication-deficient retrovirus containing the construct for beta galactosidase (BAG); BAG was deposited into the cingulum at the level of the septal nuclei. Morphological profiles, generated using Xgal substrate to visualize labeled cells, revealed a population of simple, immature cells. The antigenic profile, generated immunohistochemically with cell specific markers 2 or 30 days post injection (dpi), showed a population of cells that primarily expressed nestin or an oligodendrocyte-specific glutathione-S transferase isoform, Yp (GST-Yp) at 2 dpi and nestin, GST-Yp or Rip at 30 dpi. Occasionally, labeled cells differentiated in vivo into myelinating oligodendrocytes 30 dpi. Labeled cells did not express the astrocyte markers GFAP, GST-Yb, or S100 beta at 2 or 30 dpi. Comparisons of cell distribution 2 and 30 dpi indicated the non-migratory nature of these cells. Cell distribution patterns and nearest neighbor analyses confirmed the emergence of clusters of labeled cells 30 dpi, which bromodeoxyuridine (BrdU) incorporation studies suggested arose from continued proliferation of some labeled cells. In vivo characterization of proliferating cells in the adult revealed a non-migratory, primarily undifferentiated population of cells. PMID- 8723842 TI - Beta-adrenergic receptors regulate astrogliosis and cell proliferation in the central nervous system in vivo. AB - Astrocytes express several cell surface receptors including the beta 2 adrenergic receptor. To explore whether beta-adrenergic receptors (beta-ARs) directly regulate astrogliosis and glial scar formation, we evaluated the effects of beta-AR activation and blockade on astrocyte hypertrophy and cell proliferation in rabbit optic nerves in vivo. Artificial cerebrospinal fluid (CSF), isoproterenol (ISO; a beta-agonist), or propranolol (PROP; a beta antagonist) were infused via osmotic minipumps into non-injured and crushed optic nerves for 14 days. Changes in nerve cell numbers and astroglial hypertrophy were monitored by ethidium bromide nuclear staining and glial fibrillary acidic protein (GFAP) immunohistochemistry, respectively. In non-injured nerves infused with CSF or PROP, there were no alterations in GFAP-immunoreactivity or cell numbers compared to normal optic nerves; however, in non-injured nerves infused with ISO, there was a significant increase in both GFAP-immunoreactivity and cell number. In crushed optic nerves, there was a significant increase in both GFAP immunoreactivity and cell number compared to normal nerves, and this increase was not altered by infusion of either CSF or ISO. In contrast, PROP infusion significantly reduced the crush-induced increase in GFAP-immunofluorescence and cell number. These findings suggest that a) beta-AR activation, in the absence of injury, can promote astroglial hypertrophy and cell proliferation; b) after injury, beta-AR activation drives injury-induced astrogliosis and cell proliferation; c) astrocyte beta-ARs are maximally stimulated after neuronal injury; and d) neuronal regeneration may be influenced, both positively and negatively, through the pharmacological manipulation of glial receptors. PMID- 8723843 TI - ATP-evoked inositol phosphates formation through activation of P2U purinergic receptors in cultured astrocytes: regulation by PKC subtypes alpha, delta, and theta. AB - ATP-induced phosphoinositide (PI) hydrolysis was studied in cultured astrocytes. To characterize the P2 purinergic receptor-mediated effects of ATP, the subtype specific agonists 2-methylthio ATP (2-MeSATP), UTP, and alpha, beta-methylene ATP were compared. ATP, UTP, or 2-MeSATP induced a dose-dependent increase of inositol phosphates (IP) accumulation; alpha, beta-methylene ATP and adenosine had no effect. The order of potency was ATP > or = UTP >> 2-MeSATP. Cross desensitization experiments indicated that ATP interacted with both P2U and P2Y receptors. P2U was the predominant P2 receptor in mediating PI hydrolysis in astrocytes. The effect of ATP, UTP, or 2-MeSATP was markedly inhibited by pretreatment of cells with pertussis toxin (PTX), indicating that both P2U and P2Y receptors coupled to phospholipase C through PTX-sensitive G protein. Short term (10 min) treatment of cells with 1 microM TPA attenuated ATP, UTP, and 2 MeSATP-induced PI breakdown; however, long-term (24 h) pretreatment resulted in marked potentiation of both ATP and UTP, and restoration of 2-MeSATP responses. In a further analysis of the effect of TPA, 10 min and 1.5 h pretreatment attenuated ATP-and UTP-induced PI breakdown, but this inhibitory action was lost after 3 h of treatment. Both 6 and 24 h pretreatments resulted in a potentiation. Western blot analysis showed translocation of protein kinase C (PKC) alpha, delta, and -theta from the cytosol to the membrane following 10 min and 1.5 h treatments, and restoration to basal levels in the membrane fraction was seen after 3 h of treatment. On the other hand, partial and complete down-regulation of these three isoforms was seen after 6 and 24 h of treatment, respectively. PKC eta was translocated but not down-regulated by TPA. These results suggested that PKC alpha, -delta, and -theta, not -eta may exert tonic inhibition on P2U receptor-mediated PI turnover in unstimulated astrocytes. PMID- 8723844 TI - Potassium currents in cultured glia of the frog optic nerve. AB - The processes that participate in clearing increases in [K+]o produced by active neurons include KCl uptake, Na pump stimulation, and spatial buffering. The latter process requires glial cells to carry: 1) inward K+ currents in regions where K+ is elevated at a glial membrane potential more negative than EK; and 2) outward K+ currents at normal K+ and glial membrane potential more positive than EK (Orkand et al: J Neurophysiol 29:788, 1966). Techniques for isolation and culturing glial cells brought new possibilities for studying ionic channels involved in spatial buffering. However, they raised the question of the extent to which the properties of ionic channels are changed due to the process of culturing when glial cells are exposed to an artificial environment and deprived of direct interaction with neurons. We studied potassium currents in glial cells from the frog optic nerve that were cultured for 1-8 days. At 24-48 h, cells exhibited an inwardly rectifying Cs+ blocked current (IK(IN)) that increased in amplitude and shifted its threshold of activation to EK when [K+]o was increased from 3 to 6 or 10 mM. IK(IN), diminished after 3 days in culture and virtually disappeared after 5 days. At 24-48 h, a potassium delayed rectifier current (IKD) was relatively small but became large at 3 days, and was practically the only current present after 5 days. IKD was activated at -8.5 +/- 0.58 mV(SE, n = 48) and 58 +/- 2.2% (SE, n = 48) blocked by 20 mM tetraethylammonium. The results of this study support the idea that the inward rectifying potassium channels (Kir) are responsible for carrying K+ into glial cells whenever [K+]o increases. However, the delayed rectifier potassium channels (KD) cannot provide the pathway for outward K+ current during spatial buffering, and another mechanism must be involved in this process. Our study provides further evidence that culture conditions can greatly influence functional expression of ionic channels in glial cells. PMID- 8723845 TI - Simulation: it's a start. PMID- 8723846 TI - Anaesthesia crisis resource management training: an intimidating concept, a rewarding experience. AB - PURPOSE: This two-part study was performed to identify and address anaesthetists' concerns regarding anaesthesia simulation and to evaluate the response of practitioners to simulation-based Anaesthesia Crisis Resource Management Training (ACRM). METHODS: First, 150 survey questionnaires were distributed to participants of the Anaesthesia Practice '94 meeting in Toronto and to staff and resident anaesthetists at the Sunnybrook Health Science Centre. In the second part of the study, 35 anaesthetists from the Toronto area who participated in Anaesthesia Crisis Resource Management (ACRM) workshops at the Canadian Simulation Centre completed an anonymous exit evaluation questionnaire. RESULTS: Among staff anaesthetists (n = 42), 19% of the surveyed respondents had never heard about anaesthesia simulation, whereas all residents (n = 17) had heard of, or seen an anaesthesia simulator. Horizontal numerical scale ratings (from 1-10, with 10 being extremely supportive) indicated support for the purchase of a simulator (8.3 +/- 2.0 for staff, 9.2 +/- 1.1 for residents). Staff and residents anticipated substantial anxiety while training with a simulator (6.8 +/- 2.4 and 7.6 +/- 1.4 respectively, with 10 indicating extreme anxiety). Participants in the ACRM workshops at the Canadian Simulation Centre enjoyed the course (1.2 +/- 0.6, on a scale form 1 through 5, with 1 indicating total support and 5 representing no support), felt that it would be beneficial to most anaesthetists (1.2 +/- 0.5) and should be taken, on average, every 18 mo. CONCLUSIONS: Even though the majority of respondents have not been exposed to anaesthesia simulators, they appear to support their use in education strongly. Whereas substantial anxiety could delay the introduction of simulation based education, participants of ACRM workshops enjoy the courses and perceive them as very educational. PMID- 8723847 TI - Laryngeal mask insertion with a laryngoscope in paediatric patients. AB - PURPOSE: To assess epiglottic position after laryngeal mask airway (LMA) insertion with or without the use of a laryngoscope. METHODS: A double-blind randomized study. In 48 children an LMA (#2 for 6-20 kg, #2.5 for 20-30 kg) was inserted either blindly or with the help of a laryngoscope and its position assessed using fibreoptic endoscopy. RESULTS: An unobstructed view of the glottis, as assessed by fibrescope, was observed in 10 of 25 patients in the laryngoscope group, but only in 1 of 22 patients in the blind insertion group (P = 0.005). CONCLUSION: This technique offers an alternative when the standard technique has failed, or when LMA insertion precedes bronchoscopy or intubation via the laryngeal mask. PMID- 8723848 TI - Ketorolac does not decrease postoperative pain in elderly men after transvesical prostatectomy. AB - PURPOSE: To assess the postoperative analgesic efficacy and morphine-sparing effect of ketorolac in elderly patients. METHODS: Sixty ASA-physical status I to III men, aged 60-88 yr, undergoing transvesical prostatectomy were studied according to a randomized, placebo controlled, double-blind study protocol. A standard general anaesthetic was administered. Thirty minutes before concluding the surgical procedure either ketorolac 60 mg or an equal volume of saline was administered, im. Postoperative pain was assessed hourly for six hours using a 100 mm visual analog score (VAS) and a patient-controlled analgesia (PCA) device. RESULTS: Hourly PCA-demands, actual morphine delivered, and patient generated VAS pain scores were unaffected by the treatment modality. On conclusion of the study the total PCA morphine delivered was 11.9 mg +/- 1.38 and 10.8 mg +/- 1.52 for the saline and ketorolac groups, respectively. CONCLUSION: The intraoperative administration of ketorolac, 60 mg, im, was not associated with postoperative morphinesparing or improved analgesia in this elderly population. PMID- 8723849 TI - A double-blind comparison of ropivacaine 0.5%, 0.75%, 1.0% and bupivacaine 0.5%, injected epidurally, in patients undergoing abdominal hysterectomy. AB - PURPOSE: Ropivacaine is a new long-acting, injectable local anaesthetic currently undergoing clinical investigation world wide. It is structurally very similar to bupivacaine, but with less potential for central nervous system or cardiac toxicity. The purpose of this double-blind study was: to investigate the dose response relationship of increasing doses of ropivacaine on the quality of anaesthesia and the duration of both motor and sensory blockade, and to compare these results with an established local anaesthetic, bupivacaine. METHODS: One hundred and twenty five patients were randomly assigned to one of four treatment groups and 116 completed the study. Epidural anaesthesia was established using 25 ml test solution, injected over three minutes following a satisfactory test dose. Sensory onset, spread and duration, using the pin prick method, and motor scores using a modified Bromage scoring system were compared. RESULTS: A dose/response relationship was observed with increasing doses of ropivacaine for all variables tested except analgesia and muscle relaxation (P < 0.01). There were differences in: (i) motor onset (Levels 1 and 2), when ropivacaine 1.0% was compared with ropivacaine 0.75% and 0.5% (P < 0.05); (ii) in sensory duration at all levels except T6 when ropivacaine was compared with ropivacaine 0.5% (P < 0.05); (iii) differences in sensory duration at T12 and S1 when ropivacaine 1.0% was compared with bupivacaine 0.5% (P < 0.05); (iv) differences in motor duration at all levels when ropivacaine 1.0% was compared with ropivacaine 0.5% (P < 0.05). No serious adverse events were reported in this study. CONCLUSION: Increasing doses of ropivacaine were associated with an increased clinical effect. The most consistent differences occurred when ropivacaine 1.0% was compared with 0.5% and the least consistent between ropivacaine 0.5%, 0.75% and bupivacaine 0.5%. The main difference between ropivacaine 1.0% and bupivacaine was in sensory duration. No serious adverse events were reported. PMID- 8723850 TI - Comparative absorption kinetics of intramuscular midazolam and diazepam. AB - PURPOSE: This study investigates the rate and extent of absorption following intramuscular injection of midazolam and diazepam. METHODS: Four healthy male volunteers were recruited in this randomized three-way cross-over study. On one occasion each subject received simultaneous im injections of 5 mg midazolam and 10 mg diazepam in separate deltoid muscles. On two other separate occasions each subject received an iv infusion of 7.5 mg midazolam and 30 mg diazepam over five minutes. Frequent arterial blood samples were collected for up to two hours and venous blood samples were collected for up to 24 hours for midazolam and ten days for diazepam. A gas chromatography assay was used to determine the plasma concentrations of midazolam and diazepam. The im absorption profiles were estimated using constrained least-squares deconvolution. RESULTS: There were substantial intersubject variabilities in the estimated pharmacokinetic parameters (volume and clearances) of intravenous midazolam and diazepam. The mean (+/-sd) time to peak plasma concentration (Cmax) was shorter for im midazolam (17.5 +/- 6.5 min) relative to diazepam (33.8 +/- 7.5 min). The mean (+sd) time to peak absorption rate was also shorter for midazolam (9 +/- 2 vs 13.8 +/- 7.5 min). The peak rate of absorption was identical (0.18 mg. min-1) and bioavailability was 1.0 for both drugs. CONCLUSIONS: We conclude that midazolam has more rapid absorption than diazepam following im administration. PMID- 8723851 TI - Ulinastatin reduces elevation of cytokines and soluble adhesion molecules during cardiac surgery. AB - PURPOSE: To investigate whether ulinastatin pretreatment (6000 U.kg-1 before CPB and before declamping of aorta) influenced the production of cytokines and adhesion molecules in the peripheral circulation. METHODS: This prospective randomized study was performed in 22 patients undergoing cardiac surgery. They were divided into two groups. Patients in Group I were untreated and in Group II treated with ulinastatin. The soluble intercellular adhesion molecule-1 (S-ICAM 1), soluble endothelial leukocyte adhesion molecule-1 (S-ELAM-1), interleukin 8 and 6 (IL- 8, 6) were measured using ELISA kits. RESULTS: Serum S-ICAM-1 concentration in Group I increased from the preoperative value of 297 +/- 27 ng.kg-1 to 418 +/- 106 ng.kg-1 at 60 min after declamping of the aorta (P < 0.01) but did not change in Group II. Serum S-ELAM-1 concentration did not change in either group. Serum concentration of IL-8 and IL-6 in Group I (37 +/- 44 pg.kg-1, and 59 +/- 59 pg.kg-1, preoperatively) increased to 169 +/- 86 pg.kg-1 and 436 +/ 143 pg.kg-1 at 60 min after declamping of the aorta (P < 0.001, P < 0.001). The increases were greater than those from 25 +/- 6 pg.kg-1 and 30 +/- 26 pg.kg-1 to 56 +/- 36 pg.kg-1 and 132 +/- 78 pg.kg-1 in Group II (P < 0.001, P < 0.001). The levels of S-ICAM-1 correlated with those of IL-8 (r = 0.5, P < 0.001). CONCLUSION: These results suggest that ulinastatin may suppress the increase in IL-8 production and the expression of ICAM-1 during cardiac surgery. PMID- 8723852 TI - Cerebral blood flow velocity after mannitol infusion in children. AB - PURPOSE: There is conflicting evidence as to whether the effect of mannitol on brain bulk arises from haemodynamic, rheologic, or osmotic mechanisms. If mannitol alters cerebral haemodynamics by inducing vasoconstriction, this change should be reflected in cerebral blood flow velocity (CBFV) in the middle cerebral artery (MCA). The purpose of this study was to evaluate the effect of mannitol on CBFV in children. METHODS: Children scheduled for intracranial surgery were enrolled. After a loading dose of 10 micrograms.kg-1 of fentanyl, general anaesthesia was maintained with fentanyl (3 micrograms.kg-1.hr-1), 66% nitrous oxide, and isoflurane (0.2-0.5% inspired). Mean and systolic CBFV (Vm and Vs) and pulsatility index (PI) were recorded with a transcranial Doppler (TCD) directed at the M1 segment of the MCA. Mannitol was administered, 1 gm.kg-1 iv over 15 min. The osmolality (Osm), haematocrit (Hct), mean arterial pressure (MAP), heart rate (HR), and TCD variables were recorded before and 15, 30, 45, and 60 min after the mannitol infusion. RESULTS: Mannitol infusion resulted in an increase in Osm and decrease in Hct (P < 0.05). Heart rate, MAP and arterial carbon dioxide tensions did not change (P > 0.05) during the measuring period. The Vm did not vary from baseline. The Vs and PI both increased briefly (P < 0.01 at 15 min and P < 0.05 at 30 min) after the mannitol, suggesting an increase in resistance distal to the MCA. CONCLUSION: The time course of CBFV changes produced by mannitol corresponds with previous animal data concerning cerebrovascular tone. Our results suggest that mannitol briefly increases cerebrovascular resistance and thereby diminishes cerebral blood volume. PMID- 8723853 TI - Anaesthetic management of a patient with glycogen storage disease type 1b. AB - PURPOSE: A rare case of a ten-year old patient with type 1b glycogen storage disease (GSD), scheduled for extracorporeal shockwave lithotripsy (ESWL), is described. CLINICAL FEATURES: Patients with type 1b GSD manifest a range of clinical symptoms, including mental retardation, hepatosplenomegaly, renal enlargement, stomatitis, hypoglycaemic convulsions, bleeding diathesis, lactic acidosis and leukopaenia, thus creating a challenge for the anaesthetist. Following preanaesthetic administration of glucose-containing fluids, general anaesthesia was induced and the patient was mechanically ventilated. Except for mild hypoglycaemia after induction of anaesthesia, and moderate intraoperative metabolic acidosis which was attributed to the underlying disorder, anaesthesia was uneventful. No postoperative complications occurred and the patient was discharged home three days after lithotripsy. Clinical features of this rare inborn error of metabolism are reviewed and the approach for the anaesthetic management is discussed. CONCLUSIONS: A skillful perioperative management of patients with type 1b GSD can be achieved by cautious attention to the metabolic and homeostatic derangements that occur with the disease. PMID- 8723854 TI - Bacterial meningitis and cauda equina syndrome after epidural steroid injections. AB - PURPOSE: To describe a rare adverse outcome resulting from lumbar epidural steroid injections for the treatment of chronic lower back pain. CLINICAL FEATURES: We report a case of staphylococcus aureus meningitis and cauda equina syndrome following a series of epidural steroid injections for chronic back pain. Although rare, bacterial meningitis following epidural analgesia has been reported, but epidural steroid injections have not been associated with either bacterial meningitis or cauda equina syndrome. The causal relationship between epidural steroid injections, bacterial meningitis, and cauda equina syndrome is discussed. CONCLUSION: A through pre-procedure assessment with attention to the neurologic examination and signs/symptoms of infection is essential. PMID- 8723855 TI - Properties of fibres, endplates and acetylcholine receptors in the diaphragm, masseter, laryngeal, abdominal and limb muscles in the goat. AB - PURPOSE: Although differences in fibre composition, fibre size or acetylcholine receptor (AChR) density between muscles have often been proposed to explain the unequal sensitivities of muscles to muscle relaxant drugs, it is not clear whether or how these parameters differ among muscles or are related to one another, In this study, several muscles were examined to determine the composition and cross-sectional area (CSA) of types I and II fibres, the surface area of their motor endplates (ESA), and their AChR density. METHODS: Biopsies from the thyroarytenoideus, cricoarytenoideus dorsalis, masseter, diaphragm, transversus abdominis, rectus abdominis, gastrocnemius and soleus muscles of goats were processed by muscle histochemistry and morphometry and the ESA:CSA ratio was computed. The number and density of AChRs per endplate were estimated by 125I-alpha-bungarotoxin binding studies. RESULTS: The mean type I fibre composition (range: 0-100%), fibre diameter (28-50 microns) and the ESA:CSA ratio (0.27-1.01) differed among muscles (P = 0.0001), but there were no significant differences (P > 0.05) in the mean endplate size (577-725 microns 2) AChR number (6.6-14.5 x 10(6)) or AChR density (8,900-22,300 microns-2) probably because of marked individual variations. Fibre size increased and the ESA:CSA ratio decreased in the order laryngeal, diaphragm, jaw, limb and abdominal muscles. CONCLUSION: It is concluded that between muscles fibre size varies more than endplate size or AChR number. PMID- 8723856 TI - Duration of succinylcholine and vecuronium blockade but not potency correlates with the ratio of endplate size to fibre size in seven muscles in the goat. AB - PURPOSE: Muscles differ in their response to neuromuscular relaxants. This study investigated whether (1) the relative responses of muscles is inverted between succinylcholine (SUX) and vecuronium (VEC), and (2) differences in dose-response or duration of action are related to the morphology of fibres, endplates and acetylcholine receptors (AChR) in muscles. METHODS: In goats during thiopentone anaesthesia, the evoked EMG response to indirect train-of-four stimulation was monitored and the cumulative dose-response curves and duration of action of SUX and VEC in the diaphragm, cricoary-tenoideus dorsalis, thyroarytenoideus, transversus abdominis, rectus abdominis, soleus and gastrocnemius muscles were determined and related to their fibre composition, fibre size, endplate size, endplate to fibre size ratio, AChR number or AChR density by regression analysis. RESULTS: There were no differences in the ED50S of SUX [range, 119 +/- 11 (SE) to 159 +/- 20 micrograms.kg-1] or VEC [range, 2.8 +/- 0.2 to 3.7 +/- 0.8 microgram.kg-1] among muscles. With either drug, duration to 25% or 50% T1 recovery was shortest at the laryngedl muscles and longest at abdominal muscles (P = 0.0001), and correlated directly with fibre size (r > or = 0.40; P < 0.004) and inversely with the endplate to fibre size ratio (r > or = 0.40; P < 0.008). CONCLUSION: The results show that (1) the relative responses of muscles do not differ between depolarizing and non-depolarizing relaxants; (2) the duration of blockade is shorter in muscles composed of small fibres with large endplates relative to fibre size; and (3) there is no relation between fibre type composition and sensitivity to muscle relaxants. PMID- 8723857 TI - Isoflurane and halothane impair both systolic and diastolic function in the newborn pig. AB - PURPOSE: Volatile anaesthetics have considerable effects on diastolic relaxation in the adult myocardium. We hypothesized that isoflurane (1) and halothane (H) may have even greater effects on diastolic function in the newborn, as the newborn heart has increased passive stiffness and altered calcium handling relative to the adult. Using a newborn pig model, we compared I and H at three clinically relevant concentrations with respect to both systolic and diastolic function. METHODS: Sixteen newborn pigs were randomized for study at control (background fentanyl 100 micrograms.kg-1.hr-1 and 0.5, 1.0 and 1.5 MAC of I (n = 8) or H (n = 8). Temperature, arterial blood gases, and LVEDP were controlled. Left ventricular pressure (LVP) was monitored continuously and LV anterior posterior dimension was determined by using sonomicrometry crystals. Systolic function was assessed by peak positive dP/dT (dP/dTmax) and the slope of the end systolic pressure-dimension (ESP-D) relationship. Diastolic relaxation was given by peak negative dP/dT (-dP/dTmax) and the time constant for ventricular relaxation (tau). Left ventricular stiffness was calculated from the slope of the end-diastolic pressure-dimension (EDP-D) relationship. RESULTS: At equal MAC concentrations, I and H were identical in effect for every variable studied. Systolic function was depressed at all anaesthetic concentrations. Control dP/dTmax (I:1815 +/- 561 (SD) mmHg.sec-1, H:1841 +/- 509) decreased to 832 +/- 341 with 1.5 MAC I and 691 +/- 127 with 1.5 MAC H (P < 0.05 vs control). ESP-D slope decreased from 62 +/- 31 mmHg.mm-1 at control to 15 +/- 11 with 1.5 MAC I and from 79 +/- 16 to 37 +/- 15 with 1.5 MAC H (P < 0.05 vs control). Diastolic function was affected only at higher MAC anaesthesia. Control tau increased from 18.0 +/- 6 msec to 29.1 +/- 7.5 with 1.5 MAC I and from 20.8 +/- 5.9 to 30.0 +/- 11.3 with 1.5 MAC H (P < 0.05). EDP-D slope was increased at both 1 and 1.5 MAC anaesthesia. EDP-D slope increased from 0.16 +/- 0.24 mmHg.mm-1 at control to 0.58 +/- 0.46 with I MAC I and from 0.16 +/- 11 to 0.50 +/- 0.35 with 1 MAC H. The -dP/dTmax decreased at every MAC level of anaesthesia. CONCLUSION: These combined systolic and diastolic effects help to explain the increased sensitivity of the newborn myocardium to volatile anaesthetics. PMID- 8723859 TI - The precipitate formed by thiopentone and vecuronium. AB - PURPOSE: To determine the composition and solubility of the precipitate formed by thiopentone and vecuronium in vitro. METHODS: The precipitate formed by mixing thiopentone 2.5% and vecuronium 0.1% at room temperature was analyzed by ultraviolet spectrophotometry and high performance liquid chromatography (HPLC). The solubility of the precipitate in human plasma was measured by HPLC. RESULTS: The UV absorption spectrum of the precipitate resembled that of thiopentone. HPLC analysis produced a single peak with the same retention time as thiopentone (4.6 min). In human plasma the solubility of the precipitate was not different from that of thiopentone acid. The solubility of thiopentone was greater than that of the precipitate. CONCLUSION: The precipitate formed by thiopentone and vecuronium in vitro consisted of thiopentone acid, which was insoluble in human plasma. PMID- 8723858 TI - Thiopentone induced enhancement of somatic motor responses to noxious stimulation: influence of GABAA receptor modulation. AB - PURPOSE: This study was conducted to determine whether hyperalgesic effects of subanaesthetic concentrations of thiopentone could be attributed to GABAA receptor effects. METHODS: All studies were performed on 50 rats in a prospective, randomized, blinded fashion using saline-injected animals as controls. Using a modified Randall-Selitto technique, the motor behavior stimulated by noxious stimulation was quantified by determining the lowest tail pressure required to provoke a withdrawal response (somatic motor response threshold, SMRT). In the first protocol (21 rats), we studied the effects of 0.5, 1.5 and 5 mg.kg-1 i.v. of the GABAA agonist, muscimol, on SMRT. In the second protocol (20 rats), the effects of administration of saline, muscimol 0.5 mg.kg 1, or the competitive GABAA antagonist, bicuculline 0.25 mg.kg-1, upon the SMRT reducing effects of a standardized thiopentone infusion were observed. RESULTS: No dose of muscimol produced hyperalgesia. The highest dose of muscimol used (5 mg.kg-1) produced pronounced analgesic effects, raising the SMRT above 750 g. No change in SMRT was detected with the smaller doses of muscimol. Given in combination with muscimol (0.5 mg.kg-1), thiopentone produced analgesia, as shown by an increase in SMRT (P = 0.009). In the bicuculline treated animals, SMRT decreased linearly with increasing plasma thiopentone concentrations (P < 0.001). The slope of the relationship in the bicuculine group was not significantly different from that observed in the saline-treated group, indicating that bicuculline did not block the hyperalgesic effects of thiopentone. CONCLUSION: The results of these studies suggest that hyperalgesia associated with thiopentone is not mediated primarily by GABAA receptors. PMID- 8723860 TI - Cuff bulk of tracheal tubes in adolescence. AB - PURPOSE: To investigate cuff types for potential use in late childhood and early adolescence for appropriate fit. METHOD: Formulae suggest the use of a tracheal tube having a 6.5 mm internal diameter at ages 8 to 10 years, so a model system was used to test the passage of cuffed and uncuffed tracheal tubes of 6.5 mm internal diameter through calibrated holes 8.1-16.6 mm diameter, in plastic plates. RESULTS: Uncuffed tracheal tubes passed through 9.0 mm diameter holes. "High pressure" cuffed tubes passed through 10.5 to 11.3 mm holes. "Low pressure" cuffed tubes passed through 12.3 to 16.4 mm. CONCLUSION: Patients aged 8 to 10 yr have a limiting internal diameter at the cricoid of 8 to 10 mm. Their unstretched tracheal diameter below the cricoid is only about 11 mm. The actual outside diameters of "low pressure" cuffs chosen by commonly used formulae considerably exceed the limiting diameters of eight to ten year old patients, even considering recommended compensation factors. Analysis of background data indicates that these problems are especially aggravated at this age by non-linear growth of the cricoid. PMID- 8723861 TI - The contribution of Canadian anaesthetists to the evolution of cardiac surgery. AB - PURPOSE: There have been great improvements in the management of patients with heart disease over the past 50 yr much of which has been due to the development of surgical procedures for the correction of acquired and congenital cardiac abnormalities. A great deal has been written about the surgeons and the innovative procedures they developed. They were undoubtedly courageous, imaginative, knowledgeable and skillful. Little is written about the anaesthetists who often worked in the laboratory with the surgeons and provided anaesthesia for patients having this surgery which, in the early days, was experimental. The purpose of this article is to present the contributions made by Canadian anaesthetists to the evolution of cardiac surgery. PRINCIPAL FINDINGS: The contributions have been important over five clearly discernable eras and have been identified through publications. Canadian anaesthetists wrote about their experience giving anaesthesia for mitral commissurotomy, relief of pulmonic stenosis, ligation of patent ductus arteriosus, resection of aortic coarctation, correction of simple congenital heart defects under hypothermia and myocardial revascularization. When open heart surgery was introduced, Canadian anaesthetists working both in the United States and Canada were amongst the first to publish on surgery supported by cardiopulmonary bypass and anaesthesia for these procedures. CONCLUSION: An analysis of the literature and personal, verbal and written communications with anaesthetists who experienced the trials and tribulations of anaesthesia for these early surgical procedures clearly indicates that Canadians were at the forefront in advancing anaesthesia for cardiac surgery. PMID- 8723862 TI - Divided doses of methohexitone improves ECT outcome. PMID- 8723863 TI - Midazolam for caudal analgesia in children. PMID- 8723864 TI - Propofol myoclonus. PMID- 8723865 TI - Leak reduction with the LMA. PMID- 8723866 TI - Spinal ketamine anaesthesia for hemithyroidectomy. PMID- 8723867 TI - Inhibition of the protease activity in tracheobronchial aspirates of horses with chronic obstructive pulmonary disease. AB - OBJECTIVES: To clarify the role of proteolytic enzymes in the pathogenesis of chronic obstructive pulmonary disease (COPD) in horses, and to investigate new possibilities for treatment of this disease by interfering in the proteolytic process. DESIGN: Effect of antiproteolytic activity of selected protease inhibitors on tracheal aspirates was studied in vitro, and the inhibition profiles were compared with those of purified proteases. SAMPLE POPULATION: Respiratory tract secretions with antiproteolytic activity from 9 horses with COPD. PROCEDURE: Caseinolytic agar-diffusion assay. RESULTS: The protease inhibition profile of tracheal aspirates differed from horse to horse. The profiles did not resemble that of any of the pure proteases. Acetylcysteine, pentamidine, and diminazene were most effective in inhibiting proteolytic activity in tracheal aspirates in vitro. CONCLUSIONS: A mixed type of proteolytic activity is present in the respiratory tract secretions of horses with COPD. CLINICAL RELEVANCE: Acetylcysteine, pentamidine, and diminazene seem to have potential to be used in vivo to protect the lungs of horses with COPD from proteolytic damage. PMID- 8723868 TI - Comparison of the abilities of serologic tests to detect pseudorabies-infected pigs during the latent phase of infection. AB - OBJECTIVE: To compare the sensitivities of all available serologic tests in detecting pseudorabies virus (PRV) antibodies in pigs during long-term latent pseudorabies. DESIGN: Pigs experimentally infected with a virulent strain of PRV were maintained for 2 to 27 months after inoculation. At the time of necropsy of each pig, blood was collected for serologic evaluation, and tissues were obtained for polymerase chain reaction (PCR) verification of latency. ANIMALS: 65 crossbred pigs each weighing approximately 18 kg at the start of the study. PROCEDURE: Serum samples from each pig were analyzed by serum neutralization, latex agglutination, screening ELISA, particle concentration fluorescence immunoassay, automated latex agglutination, and differential ELISA for glycoproteins I, III, and X. DNA was extracted from the trigeminal ganglia and tonsils of each pig and was analyzed by PCR for PRV genomic sequences. RESULTS: PCR analysis of trigeminal ganglia and tonsils indicated that all pigs were latently infected with PRV at the time of necropsy, and serologic testing verified that all pigs had PRV-specific antibodies, regardless of duration of infection. The screening tests were virtually equivalent in sensitivity for detection of PRV antibodies. Of the differential serologic tests, the glycoprotein-I and -III marker systems, which performed with similar sensitivity as screening tests, were superior to the glycoprotein-X marker system in detecting PRV antibodies in latently infected pigs. CONCLUSION: Serologic testing consistently detects pigs in the latent phase of PRV infection, provided that the test detects the antibody response to the whole virus or to a reliable PRV-marker glycoprotein. PMID- 8723869 TI - Benign familial hyperphosphatasemia in Siberian huskies. AB - OBJECTIVE: To evaluate benign familial hyperphosphatasemia involving serum alkaline phosphatase (SAP) in pups. DESIGN: Pups with markedly increased SAP activity were evaluated and compared with unaffected siblings, and with other unaffected Siberian Husky pups from the same colony. ANIMALS: 8 related litters of Siberian Husky pups (n = 56). PROCEDURE: At ages 11 and 16 weeks, pups were given physical examinations and blood was obtained for hematologic and serum biochemical analyses (including electrolytes and isoenzymes of alkaline phosphatase), ionized calcium concentration, and serum parathyroid hormone concentration. Diet, growth and health performance, skeletal radiographs, and genealogical data also were evaluated. RESULTS: Of 42 pups tested, 17 had markedly high total SAP values. Mean total SAP activity of affected pups at ages 11 and 16 weeks was over 5 times greater than mean total SAP activity of unaffected siblings and other unaffected Siberian Husky pups of the same age (P < 0.001). Clinical, radiologic, and biochemical evaluation of the subjects revealed no other abnormal findings. The source of the increased SAP activity was characterized in 5 affected pups as bone isoenzyme. The mode of inheritance could be deduced from the data, but the trait clearly is familial and autosomal. CONCLUSION: The condition described in the family of Siberian Huskies bears similarity to human benign, persistent, familial hyperphosphatasemia. CLINICAL RELEVANCE: Benign familial hyperphosphatasemia should be considered in the differential diagnosis of markedly increased SAP activity in young dogs. PMID- 8723870 TI - Comparison of hematologic reference intervals between culture system and type of hybrid striped bass. AB - OBJECTIVE: To determine and compare hematologic reference intervals for hybrid striped bass from different culture systems and for 2 types of hybrid. DESIGN: Observational comparison study. ANIMALS: Clinically normal sunshine bass (reciprocal hybrid striped bass, female Morone chrysops X male M saxatilis) raised in high-density recirculating systems, sunshine bass raised in tanks with biofilters, and palmetto bass (original hybrid striped bass, female M saxatilis X male M chrysops) raised in tanks with biofilters. PROCEDURES: Hematologic reference intervals were determined for fish from the different production systems, following the National Committee for Clinical Laboratory Standards guidelines. Reference intervals from the 3 groups were compared. RESULTS: Reference intervals were significantly different between sunshine bass in the 2 culture systems and between sunshine and palmetto bass. Many of the differences were slight, but notable differences were observed. Sunshine bass in recirculating systems had higher total leukocyte, lymphocyte, neutrophil, and monocyte counts than did either hybrid in tanks. Palmetto bass had a greater number of neutrophils than monocytes, whereas sunshine bass had more monocytes than neutrophils. Additionally, palmetto bass had a lower PCV and hemoglobin value than did either group of sunshine bass. CONCLUSIONS: Separate reference intervals should be developed for hybrid striped bass in different culture systems and for different types of hybrids. CLINICAL RELEVANCE: Determining reference intervals for hybrid striped bass provides a tool to assess the health status of these fish. PMID- 8723871 TI - Blood biochemical reference intervals for sunshine bass (Morone chrysops x Morone saxatilis) in three culture systems. AB - OBJECTIVE: To determine and compare biochemical reference intervals for sunshine bass (hybrid striped bass) from 3 culture systems. DESIGN: Observational comparison study. ANIMALS: Clinically normal sunshine bass (reciprocal hybrid striped bass, female Morone chrysops x male M saxatilis raised in high-density recirculating systems (80 g/L), low-density tanks (5 g/L) with biofilters, and cages (70 g/L) in a fresh water pond. PROCEDURE: Biochemical reference intervals were determined for fish from the different production systems following the National Committee for Clinical Laboratory Standards guidelines. Reference intervals from the 3 groups were compared. RESULTS: Reference intervals were significantly different between sunshine bass in the various culture systems. Though most of the differences in reference intervals were minor, fish in the high-density recirculating system had higher concentrations of total protein, albumin, globulin, creatinine, and phosphorus, and lower chloride values. There were no significant differences in glucose concentrations among the 3 groups of fish, and no differences in cortisol concentrations between fish in tanks and cages. CONCLUSIONS: Separate reference intervals should be developed for hybrid striped bass in different culture systems. CLINICAL RELEVANCE: Determining biochemical reference intervals for hybrid striped bass provides a tool to assess the health status of these fish. PMID- 8723872 TI - Quantitative investigation of population persistence and recurrent outbreaks of bovine respiratory syncytial virus on dairy farms. AB - OBJECTIVE: To determine if transmission of virus among seropositive cattle is a plausible mechanism for the permanent presence of bovine respiratory syncytial virus (BRSV) in dairy herds, and how likely, with the scenario for persistence, there will be only 1 clinical outbreak of BRSV per year. DESIGN: Build a stochastic model, parameter estimation from serologic data on BRSV, and interpret the estimated parameter values from the model analysis. SAMPLE POPULATION: Monthly data on the prevalence of antibodies directed against BRSV in sera from all cattle in 6 dairy herds. PROCEDURE: Parameter estimation applying general linear models, model analysis using calculation of the reproduction ratio for simplified models, and Monte-Carlo simulation for the whole model. RESULTS: Persistence of BRSV by transmission among seropositive cattle given estimated parameter values would be accompanied by frequent extinctions (once every 10 to 50 years) and long infectious periods in seropositive cattle (100 days). Moreover, in the model, a single clinical outbreak among seronegative cattle only occurred with external forcing. CONCLUSIONS: From these data, transmission among seropositive cattle is not a plausible mechanism for persistence of BRSV in dairy herds. PMID- 8723873 TI - Event-time analysis of the effect of 60-day milk production on the parturition-to conception interval in dairy cows. AB - OBJECTIVE: To quantify the effect of early-lactation milk and milk fat production on time to conception. DESIGN: Event-time analysis (survival analysis). SAMPLE POPULATION: 44,450 cows delivering calves between September 1985 and September 1986, from 6,227 Ayrshire dairy herds in 80 Finnish communities. PROCEDURE: Primiparous and multiparous cows were analyzed in separate Cox proportional hazards models. 60-day milk and milk fat production were categorized and regressed against days from parturition to conception for the period 56 to 120 postpartum days, controlling for season of parturition, parity, herd production, occurrence of 43 diseases, and community. RESULTS: Multiparous cows producing above the 80th percentile for 60-day milk, but with milk fat production below the 75th percentile, were significantly more likely to conceive, compared with cows with lower production and compared with cows with similar milk production but with milk fat between the 75th and 97th percentiles. They were slightly more likely to conceive than cows with highest milk fat production, although not significantly so. Primiparous cows producing above the 80th percentile for 60-day milk with milk fat below the 75th percentile also had the highest conception probability. The highest producing primiparae had significantly lower conception probability than all others. CONCLUSIONS: Controlling for disease occurrence, calving season, parity, herd production level, and community, there appeared to be an inverted U-shaped relation between 60-day production and conception probability. CLINICAL RELEVANCE: Primiparous cows that are high producers may not be receiving the same careful management that high-producing multiparous cows receive. PMID- 8723874 TI - Event-time analysis of the effect of season of parturition, parity, and concurrent disease on parturition-to-conception interval in dairy cows. AB - OBJECTIVE: To quantify the effect of season of parturition, parity, and various diseases on time to conception. DESIGN: Event-time analysis (survival analysis). SAMPLE POPULATION: 44,450 cows delivering calves between September 1985 and September 1986, from 6,227 Ayrshire dairy herds in 80 Finnish communities. PROCEDURES: Primiparous and multiparous cows were analyzed in separate Cox proportional hazards models for postpartum days 56 to 120. Occurrence of 43 diseases or disorders was recorded; codes were condensed into 25 variables offered to each model. Season of parturition was categorized into spring, summer, and fall-winter; for multiparous cows, 3 categorical variables represented parity. Models controlled for 60-day milk and milk fat production, herd milk production, and community. For all observations, parturition-to-conception interval was plotted against calendar day of parturition. RESULTS: For multiparous cows, parturition in the spring or summer and being of parity 2 or 3 4 (vs older) increased the chance of conceiving; 10 diseases or disorders decreased this probability. Similarly, in the model for primiparae, parturition in spring or summer increased the probability of conception, and 6 disorders decreased it. Disorders that were detrimental in both models were anestrus, ovulatory dysfunction, other infertility, late metritis, and clinical ketosis. CONCLUSIONS: The effect of season on the parturition-to-conception interval was marked at this latitude. Parturition during late April, causing confluence of postpartum day 55 with the summer solstice, was associated with the shortest parturition-to-conception intervals. Ketosis was found to be an important factor in lengthening the parturition-to-conception interval, highlighting the effect of negative energy balance on postpartum restoration of reproductive function. PMID- 8723875 TI - Haptoglobin response to clinical respiratory tract disease in feedlot cattle. AB - OBJECTIVE: To quantify haptoglobin response to respiratory tract disease in feedlot cattle, and to investigate its ability to predict disease outcome and response to antibiotic treatment. DESIGN: Randomized clinical trial. ANIMALS: 60 feedlot calves with clinical respiratory tract disease. PROCEDURE: Calves were randomly assigned to receive a standard antibiotic treatment regimen (TRT), or to observation pens without antibiotic treatment. Serum haptoglobin concentration was measured at initial and final examinations. Calves were examined for presence of gross pulmonary lesions at slaughter. RESULTS: Mean +/- SD serum haptoglobin concentration at initial examination was 67 +/- 108 mg/dl, with range of 0 to 508 mg/dl. Haptoglobin concentration at initial examination was similar for the TRT group and the group that did not receive antibiotic treatment, but at final examination, TRT-group calves had lower (P < 0.01) mean values. Calves receiving antibiotic treatment had haptoglobin concentration at or near zero at final examination. Calves not receiving antibiotic treatment had only slightly lower mean haptoglobin concentration at final examination, compared with initial examination. Within treatment groups, haptoglobin concentration was similar for cases with different outcomes. Calves with gross pulmonary lesions at slaughter had numerically higher, although statistically similar, haptoglobin concentrations at initial examination, compared with calves without lesions. CONCLUSIONS: Feedlot cattle with clinical respiratory tract disease have a large and variable haptoglobin response. Antibiotic treatment resulted in lower serum haptoglobin values, although low values were not required for full clinical recovery. CLINICAL RELEVANCE: Serum haptoglobin concentration may be an indicator of response to antibiotic therapy, although it appears to be unrelated to case severity or need for treatment. PMID- 8723876 TI - Mutation segregation and rapid carrier detection of X-linked muscular dystrophy in dogs. AB - OBJECTIVES: To use exon 7-specific genomic polymerase chain reaction (PCR) products to identify the genotypes of normal, affected, and carrier female dogs in pedigrees segregating Golden Retriever muscular dystrophy (GRMD), and to confirm the concordant segregation of the mutation in all carrier and affected dogs presently available. DESIGN: The GRMD mutation is found in the consensus splice acceptor site in intron 6 of the canine dystrophin gene. PCR cycle sequencing and restriction fragment length polymorphism/PCR were used for determination of the pattern of segregation of the point mutation which causes GRMD. ANIMALS: Normal, clinically affected, and obligate carrier dogs in pedigrees of GRMD. PROCEDURE: DNA from blood was amplified, using PCR and primers that bracket all of exon 7 of the canine dystrophin gene as well as 100 base pairs of intron on either side. PCR products were either cycle-sequenced directly or submitted to a second round of PCR, using 1 of the original primers coupled with a mutagenic restriction fragment length polymorphism-primer, which thus creates an artificial restriction site. Digestion with Stu I detected the normal allele. To detect the affected allele, Sau96 I was used to digest the 310-base pair exon 7 genomic fragment directly. CONCLUSIONS: Simple, clear diagnosis of carrier status was possible using these methods. This mutation is passed through all carrier and affected dogs in both United States GRMD colonies and the colony in Australia. CLINICAL RELEVANCE: Rapid, accurate diagnosis of carrier and affected dogs will enhance study of this homologue of Duchenne muscular dystrophy. PMID- 8723877 TI - Effect of vaccination of ponies with A4 anti-idiotypic antibody on serum idiotype (1C9) and antilipid A concentration. AB - OBJECTIVE: To evaluate the humoral response of horses to vaccination, using a murine monoclonal anti-idiotypic antibody (A4) that shares an epitope with lipid A. DESIGN: Serum concentrations of antilipid A antibody and 1C9 (epitope on murine monoclonal antilipid A antibody) were measured serially during the period of vaccination with A4. ANIMALS: 6 clinically normal adult ponies. PROCEDURE: Ponies were inoculated IM 3 times at monthly intervals with A4. Two weeks after each inoculation, serum was obtained and was assayed by ELISA for antilipid A and 1C9 concentrations. Additional vaccinations were given to 2 of the ponies after a several-month rest period. RESULTS: There was significant increase in 1C9 concentration (P < 0.0001) during the period of vaccination and a trend toward increased antilipid A concentration. The latter effect was not significant (P = 0.055). Additional vaccinations produced further increase in serum 1C9 concentration; antilipid A concentration increased in one of these ponies but not the other. Maximal antilipid A concentration recorded in these ponies was approximately 6 times preimmune concentration and was comparable to that found in a commercial antiendotoxin antiserum. 1C9 also was detected in the commercial antiserum. CONCLUSIONS: A4 anti-idiotype vaccination of horses is safe and may be effective in eliciting an antibody response against endotoxin. The finding of 1C9 in a commercial antiendotoxin antiserum indicates that this idiotype may be part of the normal polyclonal antibody response of horses to endotoxin. CLINICAL RELEVANCE: It may be possible to use anti-idiotypic antibody vaccination in horses to induce protection against the effects of endotoxin. PMID- 8723878 TI - Attempt to pharmacologically modulate procoagulant activity of lipopolysaccharide stimulated adherent bovine alveolar macrophages. AB - OBJECTIVE: To investigate the effects of anti-inflammatory drugs on lipopolysaccharide-induced procoagulant activity of bovine alveolar macrophages. DESIGN: Procoagulant activity was induced in bovine alveolar macrophages from 4 healthy Holstein calves aged 6 to 16 weeks by incubation with lipopolysaccharide. 3 anti-inflammatory drugs were used at 4 concentrations and 3 times to pretreat the alveolar macrophages. Results were analyzed to determine whether drug, concentration, or exposure period had a significant (P > 0.05) effect. PROCEDURE: Bovine alveolar macrophages, harvested by volume-controlled bronchoalveolar lavage, were pretreated for 30, 60, or 120 minutes with an anti-inflammatory compound (dexamethasone, flunixin meglumine, or phenylbutazone) at several concentrations ( 0, 1, 10, and 100 microM). Bovine alveolar macrophages were exposed to lipopolysaccharide (Escherichia coli O55:B5) in the presence and absence of fetal bovine serum for 4 hours. Procoagulant activity was measured, using a chromogenic assay. RESULTS: None of the drugs was associated with a modification of procoagulant activity expression. CONCLUSION: Use of these 3 anti inflammatory drugs is unlikely to modify the extent of the fibrinous reaction commonly observed in cases of acute bovine respiratory tract disease complex. CLINICAL RELEVANCE: The alveolar macrophage has a key role in fibrin production. Assuming in vivo events mimic the in vitro model, is appears unlikely that administration of anti-inflammatory drugs will reduce the procoagulant activity of the bovine alveolar macrophages and the directly associated pulmonary fibrosis. PMID- 8723879 TI - Contribution of passive immunity to porcine respiratory coronavirus to protection against transmissible gastroenteritis virus challenge exposure in suckling pigs. AB - OBJECTIVE: To determine the ability of porcine respiratory coronavirus (PRCV) infections to induce passive immunity in suckling pigs to transmissible gastroenteritis virus (TGEV) challenge exposure. DESIGN AND ANIMALS: 4 TGEV seronegative sows and their litters (group A) served as controls, whereas 2 other groups (B and C) of sows (also TGEV seronegative) were oronasally inoculated with live PRCV during 1 or 2 subsequent pregnancies, respectively. PROCEDURE: Effectiveness of passive immunity provided to pigs via colostrum and milk was assessed after TGEV challenge exposure, and TGEV antibody responses in colostrum and milk were analyzed. RESULTS: Mortality in the 3 groups of young pigs correlated with severity of clinical signs of TGEV infection and was highest in control litters (86% in group-A pigs) and lowest in litters of sows inoculated with PRCV in 2 subsequent pregnancies (14% in group-C pigs). Virus-neutralization and IgA and IgG TGEV antibody titers of milk collected from sows at challenge exposure had significant positive correlation with litter survival. Significantly higher numbers of TGEV-specific IgA and IgG antibody-secreting cells were found in group-A pigs than in group-C pigs, suggesting that high titer of maternal antibodies (induced in group-C sows multiply exposed to PRCV) may interfere with active antibody responses. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that, in PRCV-infected pig herds, multiple exposures of pregnant sows are associated with higher IgA and IgG antibody titers to TGEV in milk, and these titers contribute to protection against TGEV infection. PMID- 8723880 TI - Identification of neutrophil NADPH oxidase proteins gp91-phox, p22-phox, p67 phox, and p47-phox in mammalian species. AB - OBJECTIVE: To identify and characterize 4 components of the NADPH oxidase complex, gp91-phox, p22-phox, p67-phox, and p47-phox, in common laboratory animal species. ANIMALS: 2 clinically normal animals from each of the following species: rabbit, sheep, cow, pig, and macaque (Macaca nemistrina). A pool of 8 rats. PROCEDURE: Neutrophils were harvested from blood, Membrane and cytosol fractions were isolated and separated by sodium dodecyl sulfate-polyacrylamide gels. Gels were transferred, and immunoblots were probed with antibodies directed against individual human NADPH oxidase proteins. Human neutrophil membrane and cytosol fractions served as controls. RESULTS: Immunoreactive bands were observed in all species for gp91-phox, p67-phox, and p47-phox proteins. Immunoreactive bands for p22-phox protein were observed in cells from rats, rabbits, pigs, and macaques. CONCLUSIONS: The NADPH oxidase and its component proteins have been highly conserved across mammalian species. Lack of immunoreactivity to p22-phox in sheep and cows can be explained by sequence divergence and epitope variability at the p22-phox C-terminus. CLINICAL RELEVANCE: The high degree of NADPH oxidase protein conservation may allow the existing knowledge of the human neutrophil NADPH oxidase to be applied to the study of animal disease. PMID- 8723881 TI - Protection of BALB/c mice against homologous and heterologous species of Brucella by rough strain vaccines derived from Brucella melitensis and Brucella suis biovar 4. AB - OBJECTIVE: To evaluate stable rough mutants derived from Brucella melitensis 16M and B suis 2579 (biovar 4) as vaccines against homologous and heterologous Brucella spp in the BALB/c mouse model. DESIGN, ANIMALS, AND PROCEDURE: Rough mutants VTRM1 and VTRS1 were obtained from B melitensis 16M and B suis 2579, respectively, by allelic exchange of rfbU gene encoding mannosyltransferase with a Tn5-disrupted rfbU gene. Mice were vaccinated with VTRM1 or VTRS1 and challenge exposed 8 weeks later. RESULTS: VTRM1 and VTRS1 replicated extensively in the spleen during the first 3 weeks of infection, then decreased rapidly. Antibodies specific for the O polysaccharide were not detected in sera of mice inoculated with either rough strain. Vaccination with VTRM1 or VTRS1 induced protection against virulent strains of B abortus (2308), B melitensis (16M), B suis biovar 1 (750), and B suis biovar 4 (2579). VTRM1 also protected against B ovis (PA) and against 4 field isolates of B abortus from bison or elk. VTRS1 conferred protection against 4 field isolates of B suis biovar 4 from reindeer. Vaccines prepared from live VTRM1 or VTRS1 provided significantly greater protection than that afforded by vaccines of killed cells in QS-21 adjuvant. Vaccination with VTRM1 containing VTRS1 gave minimal protection against the antigenically unrelated Listeria monocytogenes, thus demonstrating the immunologic specificity of protection against Brucella spp. CONCLUSIONS AND CLINICAL RELEVANCE: Results encourage evaluation, in primary host species, of VTRM1 and VTRS1, along with RB51, as alternative vaccines to strain 19, Rev 1, or other smooth phase vaccines. PMID- 8723882 TI - Venereal shedding of ovine lentivirus in infected rams. AB - OBJECTIVE: To assess shedding of ovine lentivirus (OvLV) in semen of infected rams with or without epididymitis. DESIGN: Rams 1 and 2 were naturally infected with OvLV. Rams 3-6 were inoculated with OvLV strain 85/ 34. Ram 7 was inoculated with uninfected cell culture supernatant (OvLV-negative control). 14 weeks after OvLV inoculation, rams 1-3, 6, and 7 were inoculated with Brucella ovis into the epididymis. Ram 4 was a natural case of B ovis epididymitis, and ram 5 was left noninoculated (B ovis-negative control). Blood mononuclear cells (BMNC) and semen were collected between 0 and 44 weeks after OvLV inoculation. ANIMALS: Seven 2- to 3-year-old rams. PROCEDURE: Infective OvLV in BMNC and semen was determined by virus isolation and subsequent OvLV-DNA amplification by polymerase chain reaction (PCR). Bronchoalveolar lavage cells collected after death were used for DNA extraction and PCR amplification. RESULTS: OvLV was detected in the semen of rams 3 and 6, but only after B ovis inoculation. OvLV was isolated consistently from BMNC of rams 3 and 6, but only occasionally from rams 1, 2, 4, and 5. Leukocytospermia was evident in every ejaculate of all B ovis-infected rams after infection. Semiquantitative PCR determination of OvLV DNA from bronchoalveolar lavage cells revealed the highest OvLV DNA load in rams 3 and 6. CONCLUSIONS: Leukocytospermia and a high virus load in infected animals are important factors that determine shedding of OvLV in semen. CLINICAL RELEVANCE: Dissemination of OvLV through contaminated semen could have important implications in the epidemiology and control of this infection. PMID- 8723883 TI - Serum alpha-tocopherol, vitamin A, and blood selenium concentrations, and glutathione peroxidase activity in llamas fed alfalfa hay. AB - OBJECTIVES: To measure blood selenium concentration and glutathione peroxidase (GSH-Px) activity and serum concentrations of vitamin A and alpha-tocopherol, and to determine the correlation between blood selenium concentration and GSH-Px activity of llamas fed alfalfa hay. DESIGN: Mean (+/- SD) serum vitamin A and alpha-tocopherol concentrations, blood selenium concentrations, and GSH-Px activity were calculated from 9 sequential blood samples collected from llamas fed a diet of alfalfa hay. ANIMALS: 15 clinically normal llamas (8 males, 7 females) between 10 and 14 months of age. PROCEDURE: Llamas were fed alfalfa hay for 40 days prior to sample collection and then for the duration of the trial. Vitamin E, selenium, and concentrations of vitamin A precursors were measured in the hay. Blood samples were collected on days 0, 6, 7, 9, 13, 20, 42, 64, and 98. Blood selenium concentrations were measured, using an inductively coupled spectrometric method. Blood GSH-Px activity was measured with a spectrophotometer, using a modification of a previously described assay. Isocratic high-performance liquid chromatography with florescent detection was used to determine serum alpha-tocopherol and vitamin A concentrations. RESULTS: The alfalfa hay contained 0.2 mg/kg of selenium, 5 mg/kg of vitamin E, and 0.9 mg/kg of vitamin A precursors. The mean (+/- SD) blood selenium concentration and GSH-Px activity were 0.179 +/- 0.032 micrograms/ml and 25.76 +/- 6.53 mU NADPH oxidized/min/mg of Hb, respectively, with a correlation coefficient of 0.97. The mean (+/- SD) concentrations for serum alpha-tocopherol and vitamin A were 128.1 +/- 41.7 and 74.8 +/- 5.5 micrograms/dl, respectively. CONCLUSIONS: Blood selenium concentrations in llamas are highly correlated to GSH-Px activity. Blood selenium concentrations in llamas appear to be similar to other domestic ruminants and diets containing 0.2 mg/kg of selenium appear to provide an adequate dietary source. The concentrations of vitamin A precursors and vitamin E in the hay were below currently recommended dietary levels for llamas, and alfalfa hay appears to provide an unreliable source of vitamins A and E in this species. Further studies are required to determine optimal dietary concentrations and to substantiate a reference range for these vitamins in llamas. PMID- 8723884 TI - Expression of the oncogene c-erbB-2 in canine mammary cancers and tumor-derived cell lines. AB - OBJECTIVE: To determine, for canine mammary tumors, whether malignancy, with or without local invasion or regional metastasis, was associated with overexpression of the oncogene c-erbB-2. DESIGN: c-erbB-2 expression was measured in canine mammary tumor-derived cell lines and in mammary tumor tissues from clinical cases. Clinical samples were examined histologically to determine whether they were benign or malignant and, if malignant, whether they had evidence of local invasion or regional metastasis. Canine fibroblast cultures and normal canine mammary epithelial tissues were used as reference standards for cell lines and mammary tumors, respectively. SAMPLE POPULATIONS: 28 canine mammary tumor tissue samples obtained surgically from clinical cases and samples from 7 canine mammary tumor cell lines derived from primary canine mammary tumors. PROCEDURE: c-erbB-2 mRNA levels were determined by means of hybridization of total polysomal RNA with a 32P-labeled human c-erbB-2 probe on dot blots, and results were quantified by means of scanning densitometry. Overexpression of c-erbB-2 was defined as an autoradiographic density > or = 2 times the density of reference samples on the same blot. RESULTS: Overexpression of c-erbB-2 was detected in 17 of 23 malignant tumors, 0 of 5 benign tumors, and 2 of 7 mammary tumor cell lines. c-erbB-2 overexpression was correlated with a histopathologic diagnosis of malignancy (P = 0.005) but not with the presence of local invasion or regional metastatic disease (P = 0.621). CONCLUSIONS: Results suggest that overexpression of c-erbB-2 occurs prior to the development of metastatic disease in canine mammary tumors and plays a role in the development of malignancy. PMID- 8723885 TI - Morphometric evaluation of immunoglobulin A-containing and immunoglobulin G containing cells and T cells in duodenal mucosa from healthy dogs and from dogs with inflammatory bowel disease or nonspecific gastroenteritis. AB - OBJECTIVE: To investigate the distribution of IgA- and IgG-containing cells and T cells in the villi of duodenal mucosa from healthy dogs and from dogs with inflammatory bowel disease (IBD) of gastroenteritis. DESIGN: Case-control study. ANIMALS: 28 dogs, grouped according to clinical and histologic criteria: 11 dogs with IBD, 8 dogs with non-specific gastroenteritis, and 9 healthy dogs. PROCEDURE: Endoscopic biopsy specimens of duodenal mucosa from each dog were stained specifically for IgA and IgG heavy chains and pan T-cell (CD3) antigen, using immunoperoxidase techniques. Morphometric analysis, performed via an image analysis system, was used to count IgA- and IgG-containing cells and T cells within paired contiguous villi from each dog. RESULTS: cells were the predominant immune cell type in all groups of dogs. Significant differences in the villus distribution of IgA- and IgG-containing cells and T cells were not observed. Healthy dogs had significantly higher T-cell counts than had dogs with IBD or gastroenteritis. Dogs with nonspecific gastroenteritis had a significantly higher concentration of IgA-containing cells than the other groups of dogs had. Significant group differences for IgG-containing cells also were evident, with dogs with IBD having the lowest cell counts. CONCLUSIONS AND CLINICAL RELEVANCE: High concentrations of IgA- and IgG-containing cells and T cells in the villus lamina propria cannot be reliably used to distinguish IBD from other intestinal disorders in dogs. Evaluation of T cells may be the most discriminatory method for differentiating dogs with IBD from clinically normal dogs via examination of intestinal biopsy specimens. PMID- 8723886 TI - Effects of a combination of detomidine and butorphanol on respiratory function in horses with or without chronic obstructive pulmonary disease. AB - OBJECTIVE: To evaluate the effects of detomidine and butorphanol in combination on respiratory function in horses and to determine whether these effects are more severe in horses with pre-existing respiratory dysfunction, DESIGN: Pulmonary function testing and arterial blood gas analyses were performed before and after administration of a combination of detomidine (10 micrograms/kg of body weight, i.v.) and butorphanol (20 micrograms/kg, i.v.). ANIMALS: 5 horses with chronic obstructive pulmonary disease and 5 horses free of respiratory disease (controls). PROCEDURES: Flow rates were obtained from a pneumotachograph attached to a face mask, and esophageal pressure was measured with a catheter placed in the distal third of the esophagus. Respiratory rate, tidal volume (VT), minute ventilation (VE), maximal change in transpulmonary pressure, pulmonary resistance, and dynamic compliance were calculated by use of a pulmonary function computer. Arterial blood was collected from the transverse facial artery, and blood gas partial pressures were measured with an automated blood gas analyzer. RESULTS: The combination of detomidine and butorphanol caused significant changes with time in respiratory rate, VT, VE, PaO2, and PaCO2. Changes with time in VT, VE, and maximal change in transpulmonary pressure were significantly different between groups. CONCLUSION: The combination of detomidine and butorphanol affects respiratory function in horses, and the observed changes are affected by the presence of pre-existing respiratory dysfunction. CLINICAL RELEVANCE: Changes in respiratory function after administration of a combination of detomidine and butorphanol are not necessarily more severe in horses with pre-existing respiratory dysfunction. PMID- 8723887 TI - Pharmacokinetics of diminazene in plasma and lymph of goats. AB - OBJECTIVE: To characterize the pharmacokinetics of diminazene in plasma and pseudo-afferent lymph of East Africa X Galla goats. DESIGN: The efferent prescapular lymphatic duct of 3 goats was cannulated 8 weeks after surgical removal of the lymph node. Thereafter, 3.5 mg of diminazene base/ kg of body weight was administered to these goats and to 3 noncannulated goats. PROCEDURE: Using high-performance liquid chromatography, concentration of diminazene was determined in plasma and lymph collected up to 96 hours after treatment. RESULTS: Maximal concentrations of diminazene in plasma of noncannulated goats (median [range], 4.30 [4.28 to 5.01] micrograms/ml), plasma of cannulated goats (3.94 [2.94 to 4.06] micrograms/ml), and lymph (1.06?0.73 to 1.86] micrograms/ml) were significantly different (P < 0.05); values in lymph were considerably lower than those in plasma from noncannulated and cannulated animals. Time to reach maximal concentration did not differ significantly between lymph and plasma of noncannulated and cannulated goats. Over the first 24 hours after drug administration, concentration of diminazene in plasma of noncannulated goats was generally higher than that in lymph, but thereafter was similar. Apparent volume of distribution of diminazene in the plasma of noncannulated (2.57 [1.93 to 2.60] L/kg) and cannulated (2.30 [1.04 to 2.40] L/kg goats did not differ significantly. Penetration ratio of diminazene into lymph, compared with plasma, of cannulated goats was 1.69:1. CONCLUSIONS: Disposition of diminazene in goats is characterized by higher concentration in plasma than in lymph. However, the drug persists longer in lymph than in plasma. CLINICAL RELEVANCE: The longer persistence of diminazene in lymph than in plasma may account for the enhanced therapeutic efficacy of diminazene in the early stage, compared with later stages, of a tsetse fly-transmitted trypanosome infection. PMID- 8723888 TI - Disposition of transdermally administered fentanyl in dogs. AB - OBJECTIVE: To evaluate the disposition of fentanyl after i.v. and transdermal administrations. The hypothesis was that transdermal administration of fentanyl would result in a measurable plasma opioid concentration. DESIGN: Each dog received 2 treatments in a randomized, crossover design. ANIMALS: 6 clinically normal Beagles. PROCEDURE: 2 treatments consisting of i.v. fentanyl (50 micrograms/kg of body weight) and transdermal fentanyl (50 micrograms/h) administrations. Plasma fentanyl concentrations were measured at fixed times, and pharmacokinetic values were calculated. RESULTS: Intravenous pharmacokinetics of fentanyl was similar to those previously described in dogs and provided the distribution and clearance data necessary to calculate the rate of absorption of the transdermally administered opioid. The transdermal fentanyl patch produced average steady-state concentrations of 1.6 ng/ml. The actual rate of delivery of transdermal fentanyl was 35.7 (range, 13.7 to 49.8) micrograms/h, which represented 71.48% (range, 27.45 to 99.56%) of the theoretical rate of delivery. The mean elimination half-life of fentanyl after patch removal was 1.39 hours. CONCLUSIONS: Transdermally administered fentanyl resulted in fairly constant plasma concentrations, in the range generally considered to be analgesic, from 24 to 72 hours after application of the patch. The rate of drug delivery was less than expected, and there was substantial individual variation. CLINICAL RELEVANCE: Transdermally administered fentanyl has the potential to be a clinically useful analgesic regimen in dogs, and further evaluation of its analgesic actions and potential side effects warranted PMID- 8723889 TI - Pharmacokinetic model for cefazolin distribution during total hip arthroplasty in dogs. AB - OBJECTIVES: To compare cefazolin pharmacokinetics in serum and concentrations in tissues during total hip arthroplasty in dogs with and without hip dysplasia, and to calculate the optimal dosage of cefazolin for prophylactic use during total hip arthroplasty. ANIMALS: 10 dogs with hip dysplasia and 3 clinically normal dogs. PROCEDURE: Blood samples and tissue specimens from the coxofemoral joint capsule, acetabulum, and femur were obtained during unilateral total hip arthroplasty. Cefazolin concentrations in serum and tissue specimen supernatant were determined, using high-performance liquid chromatography, for use in pharmacokinetic analysis. Mathematical simulation of serum cefazolin concentration was used to to predict the optimal dose. RESULTS: Mean pharmacokinetic constants (SEM) were 0.146 (0.013) min-1 for alpha, 4.47 min for t1/2 alpha 0.015 (0.004) min-1 for beta, 46.83 min for t1/2 beta. Significant different was not detected for cefazolin distribution and elimination between dogs with and without hip dysplasia. Additional, significant difference was not observed in pharmacokinetic parameters describing distribution and elimination between the first and second doses of cefazolin. The predicted optimal dosage regimen was 8 mg/kg of body weight, i.v. every hour or mg/kg, i.v. every 2 hours. CLINICAL RELEVANCE: For prophylactic i.v. treatment during total hip arthroplasty, use of cefazolin at a dosage of 8 mg/kg every hour or 22 mg/kg every 2 hours should maintain serum cefazolin concentrations at least 10x the minimum inhibitory concentration for 3 to 4 hours. PMID- 8723891 TI - Cardiovascular response to exogenous serotonin in healthy calves. AB - OBJECTION: To characterize the cardiovascular response to i.v. administration of serotonin (5-hydroxytryptamine [5-HT]) in calves. ANIMALS: 5 healthy unsedated Friesian calves. PROCEDURE: 41 5-HT administrations were performed: II slow infusions (duration, 5 minutes) and 30 bolus infusions (duration, 5 seconds). Cardiovascular function values were recorded before, during, and after the infusion. RESULTS: Slow infusion of 5HT first resulted in a brief period of severe bradycardia, then in sustained tachycardia with a concomitant increase in cardiac output. Systemic initial hypotension concomitant with bradycardia, then a pressor phase associated with an increase in systemic vascular resistance, and finally, a long-lasting hypotensive phase associated with decreased systemic vascular resistance. Pulmonary hypertension was associated with increased pulmonary vascular resistance, reflecting intense pulmonary vasoconstriction. Bolus infusion at increasing dosages resulted in dose-dependent bradycardia and systemic hypotension, followed by dose-dependent systemic hypertension. Unlike with slow infusion, neither the second tachycardiac nor the third systemic hypotensive phases were evident. CONCLUSIONS: 5-HT induces dose-dependent cardiovascular responses, including a reflex response followed by pulmonary and systemic vasoconstriction, in healthy calves. CLINICAL RELEVANCE: Determining the type of serotonergic receptors responsible for these responses may help to determine whether 5-HT is involved in the mechanisms underlying brisket disease in cattle. PMID- 8723890 TI - Hemodynamic effects of medetomidine-midazolam-butorphanol and medetomidine midazolam-buprenorphine combinations and reversibility by atipamezole in dogs. AB - OBJECTIVE: To characterize the hemodynamic effects of medetomidine (1 mg/m2 of body surface area; dosage, 39 to 46 micrograms/kg of body weight, IM) and midazolam (1 mg/kg of body weight, i.v.) combined with butorphanol (0.1 mg/kg, i.v.), buprenorphine (10 micrograms/kg, i.v.) or saline solution. Reversibility of these effects by atipamezole (2.5 mg/m2; dosage, 97.5 to 115 micrograms/kg, IM) was evaluated. DESIGN: 2 treated groups and 1 control group, without repetition. ANIMALS: 15 clinically normal dogs (3 groups of 5). PROCEDURE: Medetomidine was administered at time 0; midazolam and butorphanol, buprenorphine, or saline solution at time 20; and atipamezole at time 60. Heart rate, systemic and pulmonary arterial pressures, central venous pressure, body temperature, cardiac output, and arterial and mixed venous blood gas tensions and pH were measured. Cardiac index, stroke index, systemic and pulmonary vascular resistances, and left and right stroke work indexes were calculated. RESULTS: Body temperature, heart rate, cardiac index, and stroke index were significantly decrease below baseline values in some groups. Central venous pressure, pulmonary capillary wedge pressure, and systemic vascular resistance were significantly increased above baseline in all groups. Arterial and venous PO2 and pH decreased in all groups and PCO2 increased, but these changes were more pronounced when buprenorphine was administered. Arterial pressure decreased after atipamezole administration. CONCLUSION: The combinations seemed to result in cardiorespiratory depressant effects of similar importance and most of these effects, which are related to medetomidine, were reversed by atipamezole. PMID- 8723892 TI - Determination of the lifespan of erythrocytes from greyhounds, using an in vitro biotinylation technique. AB - OBJECTIVE: To determine the RBC lifespan of Greyhounds, using an in vitro labeling technique. DESIGN: RBC from dogs were labeled with NHS-biotin and their disappearance measured over time to determine RBC lifespan. SAMPLE POPULATION: 5 Greyhounds that had been vaccinated against distemper, adenovirus 1 and 2 infections, parainfluenza, leptospirosis, parvovirus, and coronavirus infections, Bordetella bronchiseptica infection, and rabies the previous year; 3 sexually intact 14-month-old Beagles served as controls. PROCEDURE: After venipuncture for CBC, catheters were inserted in the cephalic vein of each dog. Butorphanol was then administered to achieve mild sedation and analgesia, and glycopyrrolate was administered to ensure maintenance of adequate heart rate during phlebotomy. Dogs were positioned in lateral recumbency; blood was removed via jugular venipuncture, using a standard laboratory donor blood bag containing citrate phosphate-dextrose solution. Blood was transferred aseptically into sterile polystyrene containers and NHS-biotin was added. After incubation, the labeled RBC were reinfused into the dogs and the blood was allowed to recirculate for 1 hour before the first postinfusion sample was taken. At frequent intervals, blood to be analyzed was taken by jugular venipuncture, and the percentage of labeled cells was determined by flow cytometry. RESULTS: The mean RBC lifespan of non Greyhounds was significantly longer than that of Greyhounds (104.3 +/- 2.2 days vs 53.6 +/- 6.5 days; P = 0.001). A negative linear correlation was also found between age of the Greyhounds and their RBC lifespan (P = 0.01, R2 = 0.91). CONCLUSIONS: The shorter RBC lifespan of the Greyhounds may explain the finding of macrocytosis reported in earlier work. The reason for the shorter RBC lifespan in Greyhounds may be caused by differences in Greyhound RBC membrane structure or accelerated RBC removal from the circulation. PMID- 8723893 TI - Investigation into intraerythrocytic and extraerythrocytic acid-base and electrolyte changes after long-term ammonium chloride administration in dogs. AB - OBJECTIVES: To investigate the effect of ammonium chloride-induced urine acidification on acid-base status (ABS) of blood, plasma, and erythrocytes, and to compare the diagnostic value of acid-base analysis for erythrocytes with that for blood and plasma. DESIGN: Ammonium chloride (100 mg/kg of body weight) was administered orally every 12 hours for 8 consecutive days. The ABS was determined daily in venous blood, plasma, and lysed, packed erythrocytes (erythrolysate) and in urine. In plasma and erythrocytes, concentrations of sodium Na+) and potassium (K+) were analyzed. ANIMALS: 16 clinically normal (8 treated and 8 control dogs. PROCEDURE: The acid-base analysis (standard pH, standard bicarbonate concentration, base excess, and carbon dioxide tension) in blood, plasma, and erythrocytes was determined by use of the CO2, O2 gas equilibration method. Determination of urine ABS (pH, concentrations of acids and bases, net acid-base excretion, base-acid quotient) was performed by titration methods. Determination of concentrations of Na+ and K+ in erythrocytes was performed by flame photometry, and in plasma, by use of ion-specific electrodes. RESULTS: Ammonium chloride caused metabolic acidosis in blood and plasma, but no change of ABS in erythrocytes. The concentrations of K+ in plasma and erythrocytes did not change in treated dogs; however, the concentrations of Na+ in plasma and erythrocytes decreased significantly (P < 0.05) after ammonium chloride administration. Urinary acid excretion increased significantly (P < 0.05) in treated dogs; urine pH was between 4.51 and 5.49 at all times. CONCLUSIONS: Ammonium chloride administration (100 mg/kg, PO, q 12 h) causes substantial blood and urine acidification but does not influence erythrocyte ABS. In this study, determination of erythrocyte ABS did not provide any additional benefit in diagnosing metabolic acidosis, compared with analysis of blood. PMID- 8723894 TI - Differential distribution of von Willebrand factor in canine vascular endothelium. AB - OBJECTIVE: To determine whether endothelial cell (EC) von Willebrand factor (vWf) is uniformly distributed in canine blood vessels. DESIGN: Contents of EC vWf from vascular segments was evaluated in Hautchen preparations, using immuno histochemistry. EC from femoral arteries and veins and jugular veins were grown in culture, and the intracellular content and constitutive release of vWf from these cells were measured. The amount of vWf mRNA in the cultured EC was determined. ANIMALS: Vascular segments for Hautchen preparations and EC for culture were obtained from 5 and 10 clinically normal, mixed-breed dogs, respectively. PROCEDURES: Appropriate vascular segments were removed, fixed; processed for immunohistochemistry, using a monospecific polyclonal antibody to canine vWf, and Hautchen preparations were made. Intracellular and constitutive released vWf was measured, using an ELISA, and vWf mRNA was measured by Northern blot analysis. RESULTS: Intact endothelial linings from femoral veins, jugular veins, vena cava, and pulmonary veins stained more intensely than femoral arteries, carotid arteries, aorta, and pulmonary veins. Constitutive release and intracellular content of vWf in cultured EC from femoral veins was about 30 times higher than that from femoral arterial EC, which was barely detectable. Similar differences were seen in amounts of mRNA. CONCLUSIONS: There is marked diversity in EC vWf in canine vasculature that may result from differences in vWf mRNA. CLINICAL RELEVANCE: Low amounts of vWf in canine systemic arterial EC may contribute to thromboresistance of canine arteries. PMID- 8723895 TI - Effects of potentiated chlorhexidine on bacteria and tarsocrural joints in ponies. AB - OBJECTIVES: To evaluate the bactericidal properties of chlorhexidine diacetate (CHD) after potentiation with EDTA and Tris buffer (EDTA-Tris), and to find a potentiated CHD concentration that would achieve 90 to 100% killing for all bacteria tested. ANIMALS: 6 adult ponies. PROCEDURES: Serial dilutions of CHD, CHD in EDTA-Tris and EDTA-Tris alone were evaluated for bactericidal activity against Staphylococcus aureus, Escherichia coli, and Streptococcus zooepidemicus. The tarsocrural joints of 6 ponies were lavaged with either 1 L phosphate buffered saline solution (control) or 1 L of 0.0005% CHD in EDTA-Tris. Synovial fluid was collected before lavage and on days 1,4, and 8. Synovia, cartilage, and bone with cartilage were collected on day 8 when the ponies were euthanatized. RESULTS: In vitro results indicated that 0.0005% CHD in EDTA-Tris was 90% lethal to all bacteria tested. Results of synovial fluid analysis, glycosaminoglycan analysis, and histologic examination of the synovial membrane and articular cartilage indicated that joint lavage with 0.0005% CHD in EDTA-Tris was not detrimental to the synovium or the articular cartilage of pony tarsocrural joints. Changes observed were a result of the actual lavage process, the phosphate-buffered saline solution, and hemarthrosis. CONCLUSIONS: A concentration of 0.0005% CHD in EDTA-Tris was 90% lethal to all bacteria tested. Pony tarsocrural joint lavage with 0.0005% CHD in EDTA-Tris was not detrimental to the synovium or the articular cartilage. The efficacy of 0.0005% CHD potentiated with EDTA-Tris as a potential joint lavage fluid for treatment of infectious arthritis needs to be evaluated in clinical patients. PMID- 8723896 TI - Effects of U-74389G, a novel 21-aminosteroid, on small intestinal ischemia and reperfusion injury in horses. AB - OBJECTIVE: To determine the effects of the 21-aminosteroid, U-74389G, on reperfusion of the equine jejunum, using total (TVO) and partial (PVO) vascular occlusion during the ischemic period. DESIGN: TVO: 16 healthy horses were randomly allotted to 3 groups-4 horses received the vehicle alone, 6 horses received a low dosage (3 mg/kg o body weight), and 6 horses a high dosage (10 mg/kg) of U-7438G. PVO: 10 healthy horses were randomly allotted to 2 groups--5 horses received the vehicle alone, and 5 horses received the low dosage (3 mg/kg) of U-74389G. PROCEDURES: TVO was induced for 1 hour followed by 2 hours of reperfusion. During PVO, blood flow was reduced to 20% of baseline for 2 hours, followed by 2 hours of reperfusion. For both models, either the vehicle alone or the drug was given 15 minutes prior to reperfusion. Samples were obtained before, during, and after ischemia for determination of myeloperoxidase (MPO) activity, malondealdehyde (MDA) concentration, concentration of conjugated dienes (PVO experiment only), and morphometric analysis. RESULTS: TVO: tissue concentration of MDA and MPO activity were not altered in any group by ischemia or reperfusion. During ischemia, mucosal volume and surface area were reduced. After reperfusion, no further reduction occurred. After initial decrease in submucosal volume during ischemia, there was a significant increase after reperfusion in the vehicle-only group (P < 0.05). PVO: there were no alterations in the concentration of either MDA or conjugated dienes. There was significant increase in the activity of MPO during ischemia and reperfusion (P < 0.05). These effects were similar for the vehicle-only and drug groups. During ischemia, there was a significant decrease in mucosal surface area and volume (P < 0.05), that was continued during reperfusion for the vehicle-only (P < 0.05). Submucosal volume increased during ischemia and reperfusion. CONCLUSIONS AND CLINICAL RELEVANCE: Reduced blood flow during ischemia (PVO group) caused continued loss in mucosal volume and surface area during reperfusion. At the dosage given, the 21-aminosteroid, U-74389G, was not effective in preventing continued reduction in mucosal volume and surface area after restoration of blood supply in the horses subjected to reduced blood flow. PMID- 8723897 TI - Comparative genomic hybridization imaging by the one-chip true-color CCD camera kappa CF 15 MC. AB - In comparative genomic hybridization (CGH), quantitative measurement of fluorescence intensity ratios on metaphase chromosomes is the basic method for detecting copy number changes in the test DNA. The microscope images are usually acquired by high-resolution, highly sensitive, black and white (B&W) CCD cameras. This requires subsequent recording of the different color images using appropriate filter combinations for excitation and emission. We describe an alternative approach using the one-chip true-color CCD camera Kappa CF 15 MC and an Omega triple-bandpass filter for simultaneous registration of the three dyes Texas red, FTTC, and DAPI. A detailed examination of the imaging properties of the system was performed. The camera response in the three color planes R, G, and B was evaluated, and calibration factors for image correction were calculated. An error estimate is given. A complete computer program for the image analysis of CGH experiments has been developed for an 80486 PC, using the commercially available software package Optimas as the basis for image recording. Examples that confirm the suitability of the system for ratio imaging in CGH are presented. The results were compared with others obtained from the same microscope slides using an established setup consisting of a B&W CCD camera (Photometrics) and a software program based on the TCL software package and run on a Macintosh Quadra 950. The results obtained using the two different systems were found to correspond closely. PMID- 8723898 TI - Quantitative analysis of three-dimensional distribution of AgNOR proteins during interphase in leukemic cells. AB - Acidic proteins of the nucleolar organizer regions, selectively stained by silver (AgNOR-proteins), were investigated during interphase in leukemia cells with a confocal scanning laser microscope (CSLM). Simultaneous confocal fluorescence (for specific labeling of DNA, using propidium iodide) and transmitted light microscopy combined with digital deconvolution (for the location of the AgNOR proteins in nonconfocal mode) were used. The distribution of the AgNOR proteins measured by 3D microscopy was described by their number, the volume occupation of the nucleus by the AgNOR aggregates, the distance between each AgNOR, the distance of each AgNOR to the nucleolar border, and their anisotropy. The results of the 3D analysis were compared to those obtained by conventional 2D analysis, cytogenetical analysis of metaphase nucleolar organiser regions (NORs), and cell duplication rate. The descriptive power of these 3D parameters were assessed for nine leukemic cell lines. The measurements of the 3D spatial distribution of AgNORs was a better discriminant parameter than the morphological parameters (i.e., number and volume). The 3D expression of AgNORs is also a reliable parameter for assessing proliferative activity of leukemic cells and seems to be in relation with the differentiation stage of these leukemic cells. PMID- 8723899 TI - Flow-stream waveguide for collection of perpendicular light scatter in flow cytometry. AB - We report a new physical configuration for the detection of perpendicular light scatter or fluorescence in flow cytometry when using a fluid stream in air. This configuration increases the signal-to-noise ratio and narrows the coefficient of variation for uniformly sized latex spheres when compared to using a microscope objective to collect such light. The new technique views the scattered light that is trapped within the optical waveguide that is naturally formed by the flow stream in air. One efficient and simple way to detect the light trapped within this optical waveguide is to place one end of a fiber optic, with a conically polished tip and sufficiently large-core diameter, directly into the flow stream and to place an optical detector at the fiber's other end. For perpendicular light scatter, the flow-stream waveguide achieves high collection efficiency (NA = 0.88) as well as high efficiency of optical through put due to lack of surfaces between the light scatterers and the detector. We obtained 10-fold higher signals with this technique than with a long-working-distance microscope objective. The flow-stream waveguide is also much easier to align than traditional microscope lens-based systems. PMID- 8723900 TI - Reliable flow cytometric estimation of nuclear DNA content in coffee trees. AB - Flow cytometry gave high resolution of genome size in two coffee taxa (Coffea liberica dewevrei and C. pseudozanguebariae). Propidium iodide (PI) and Petunia hybrida were used as dye and internal standard, respectively. Proportionally between the DNA content and the digitized fluorescence signal was checked. Five main results were evident. First, optimal experimental conditions were established for peak location estimation (mean or mode), staining time (at least 2 minutes), high voltage (557 V) for the photomultiplier tube (PMT), and PI concentration (333 micrograms/ml). Second, a parameterization of the effects of high voltage and PI concentration were deduced from curve fitting. Third, two biases in DNA content estimation were recorded for high voltage and PI concentration, and were minimized. Fourth, the genome sizes of C. liberica dewevrei and C. pseudozanguebariae were estimated with accuracy 2C = 1.421 +/- 0.005 pg and 2C = 1.129 +/- 0.005 pg, respectively. Fifth, between-genotype variance was emphasized within each taxon. PMID- 8723901 TI - Development of a monoclonal antibody to immuno-cytochemical analysis of the cellular localization of the peripheral benzodiazepine receptor. AB - Based on the amino acid sequence deduced from the cloned human peripheral benzodiazepine receptor (PBR) gene, monoclonal antibody (Mab 8D7) was produced against the C-terminal fragment of the receptor. Immunoblot experiments, performed against purified PBR, indicated that the antipeptide antibody recognized, under denaturing conditions, the corresponding amino acid sequence of the PBR. When mitochondrial membranes from PBR transfected yeast or from THP1 and U937 cells were used on immunoblot analysis, a high level of immunoreactivity was observed at 18 kDa, the PBR molecular mass deduced from cDNA, establishing the specificity of the antibody for the receptor. Moreover, binding experiments realized with intact mitochondria demonstrated that the immunogenic sequence was accessible to the antibody indicating that the C-terminal fragment of the PBR faces the cytosol. Using this Mab we developed a technique which allowed precise quantification of PBR density per cell. Furthermore, cellular localization studies by flow cytometric analysis and confocal microscopy on cell lines displaying different levels of PBR showed that Mab 8D7 was entirely colocalized with an antimitochondria Mab. PMID- 8723902 TI - Intracellular localization of cyclin B1 during the cell cycle in glioma cells. AB - We investigated cyclin B1 expression during the cell cycle in human glioma cells cultured under asynchronous growing condition by two cytometry techniques: flow cytometry (FCM) and laser scanning cytometry (LSC). FCM analysis revealed the specific accumulation of cyclin B1 in G2/M phase with a wide intercellular variation (Dunphy WG: Trend Cell Biol 4:202-207, 1994). It is noteworthy that LSC, which is characterized by rapid quantitative analysis followed by imaging, allows morphological observation of the intracellular distribution of cyclin B1 as a function of cell cycle position cell by cell (Hunter T: Cell 75:839-841, 1993). Cyclin B1 was virtually undetectable in cells from G0/G1 phase to mid S phase, but became visible in the cytoplasm in late S phase. As cells proceeded within G2 phase, the level of cyclin B1 rapidly increased in the perinuclear region of the cytoplasm, but cyclin B1 was still faintly present in the nucleus. Cyclin B1 appeared in the nucleus at the mitotic phase. Then the nuclear membrane was disrupted and cyclin B1 was distributed evenly in the cell. The level of cyclin B1 was maximum in metaphase. However, it abruptly degraded at the end of metaphase, and subsequently G1 cells were cyclin B1 negative. PMID- 8723903 TI - Flow cytometric study of changes in the intracellular free calcium during the cell cycle. AB - We measured the intracellular levels of free cytoplasmic calcium in different phases of the cell cycle in viable rat fibroblasts, using two parameter flow cytometric analysis with Hoechst 33342 as the DNA specific dye and Fluo-3 as the calcium sensitive dye. We studied changes in calcium levels during the G1 phase of cell cycle by arresting cells with chemical agents such as staurosporine and hydroxyurea or by density dependent arrest of cell growth. We show that levels of calcium are lowest at the beginning of G1 phase but rise steadily with its progression and culminate at the G1/S border. Our results suggest that complex changes occur in calcium levels during the process of mitotic division. During progression of S and G2 phases, calcium levels decline and increase respectively. Our results offer a new methodology to estimate intracellular calcium levels in specific phases of the cell cycle. Based upon these results we propose a general scheme representing the changes in the intracellular calcium concentration during the progression of the cell cycle. PMID- 8723905 TI - Cellular sociology applied to neuroendocrine tumors of the lung: quantitative model of neoplastic architecture. AB - This paper reports on cellular sociology, which consists of modeling tissular architecture based on graph theory. Voronoi's diagram was chosen to build the models. This diagram derives from a cell neighborhood concept and generates parameters which objectively represent tissue architecture. Minimal spanning tree (MST) is probably the more frequently used among graphs and successfully discriminates different grades of pathological process. However, Voronoi's diagram is more comprehensive and a more complete representation of architecture with the advantage of stability. The lung neuroendocrine tumor classification is far from being consensual, especially for lesions which don't fall in with typical carcinoid and small cell carcinoma groups. By comparing architectural models of 20 neuroendocrine tumors of the lung, this work supports the morphologic spectrum concept of these tumors and also supports the recently proposed concept of large-cell neuroendocrine tumors of the lung. Finally, architectural parameters separate small-cell-lung carcinomas from neuroendocrine non-small-cell lung carcinomas. PMID- 8723906 TI - Methodological aspects of using decision trees to characterise leiomyomatous tumors. AB - The aim of the present work is to present the potential uses of a classification technique labeled the "decision tree" for tumor characterisation when faced with a large number of features. The decision tree technique enables multifeature logical classification rules to be produced by determining discriminatory values for each feature selected. In this report, we propose a methodology that used decision trees to compare and evaluate the information contributed by different types of features for tumor characterisation. This methodology is able to produce a set of hypotheses related to a diagnosis and or prognosis problem. For example, hypotheses can be producted (on the basis of a set of descriptive features) to explain why tumor cases belong to a given histopathological group. To illustrate our purpose, this methodology was applied to the difficult problem of leiomyomatous tumour diagnosis. The aim was to illustrate what kind of diagnostic information can be extracted from a sample data set including 23 smooth muscle tumors (14 benign leiomyomas and 9 malignant leiomyosarcomas) described by a large set of computer-assisted, microscope-generated features. Three groups of features were used relating to: (1) ploidy level determination (10 features), (2) quantitative chromatin pattern description (15 features), and (3) immunohistochemically related antigen specificities (6 features). All these features were quantified by digital cell image analysis. The results suggest that an objective distinction between leiomyomas and leiomyosarcomas can be established by means of simple logical rules depending on only a few features among which the immunohistochemically revealed antigen expression of desmin plays a preponderant part. One of the combinations of features proposed by the methodology is interesting for pathologists, because it includes two features describing the appearance of a nucleus in terms of chromatin distribution homogeneity and density, two features widely used by pathologists in tumor grading systems. PMID- 8723904 TI - Depletion of intracellular calcium stores facilitates the influx of extracellular calcium in platelet derived growth factor stimulated A172 glioblastoma cells. AB - Calcium signaling in non-excitable cells is the consequence of calcium release from intracellular stores, at times followed by entry of extracellular calcium through the plasma membrane. To study whether entry of calcium depends upon the level of saturation of intracellular stores, we measured calcium channel opening in the plasma membrane of single confluent A172 glioblastoma cells stimulated with platelet derived growth factor (PDGF) and/or bradykinin (BK). We monitored the entry of extracellular calcium by measuring manganese quenching of Indo-1 fluorescence. PDGF raised intracellular calcium concentration ([Ca2+]i) after a dose-dependent delay (tdel) and then opened calcium channels after a dose independent delay (tch). At higher doses (> 3 nM), BK increased [Ca2+]i after a tdel approximately 0 s, and tch decreased inversely with both dose and peak [Ca2+]i. Experiments with thapsigargin (TG), BK, and PDGF indicated that BK and PDGF share intracellular Ca2+ pools that are sensitive to TG. When these stores were depleted by treatment with BK and intracellular BAPTA, tdel did not change, but tch fell to almost 0 s in PDGF stimulated cells, indicating that depletion of calcium stores affects calcium channel opening in the plasma membrane. Our data support the capacitative model for calcium channel opening and the steady-state model describing quantal Ca2+ release from intracellular stores. PMID- 8723907 TI - An improved technique for optical interferometric imaging of isolated cells. AB - We have improved the optical interferometric imaging technique that was recently used to measure local organic material concentrations in quasicylindrical cells. This allowed similar measurements for cells of arbitrary shape. The setup was used to measure the thickness of skin corneocytes. PMID- 8723908 TI - Fluorescence in situ hybridization in whole-mount Drosophila embryos. PMID- 8723909 TI - Direct analysis of in vitro transcription products from supercoiled templates containing ribozymes. PMID- 8723910 TI - Heterogeneity of DNA binding to membranes used in quantitative dot blots. PMID- 8723911 TI - Microsatellite libraries enriched for several microsatellite sequences in plants. PMID- 8723912 TI - Broad-host-range cloning and cassette vectors based on the R388 trimethoprim resistance gene. PMID- 8723913 TI - PCR-based quantitation of transfected luciferase DNA: evaluation using an Entamoeba histolytica reporter construct. PMID- 8723914 TI - LA-PCR-based quick method for the identification of genes responsible for the complementation of Saccharomyces cerevisiae mutations. PMID- 8723915 TI - Identification of introns by reverse-transcription PCR. PMID- 8723916 TI - Use of elevated reverse transcription reaction temperatures in RT-PCR. PMID- 8723917 TI - Isolation of DNA from stool and bodily fluids for PCR amplification. PMID- 8723918 TI - Simultaneous extraction of DNA and RNA from nuclease-rich pathogenic protozoan Trichomonas vaginalis. PMID- 8723919 TI - Reducing "double sequences" in automated DNA sequencing with T7 DNA polymerase and internal labeling. PMID- 8723920 TI - Lysis buffer composition dramatically affects extraction of phosphotyrosine containing proteins. PMID- 8723921 TI - Improved technique for the preparation of water-in-oil emulsions containing protein antigens. PMID- 8723922 TI - Robotic automation of Xenopus oocyte bath perfusion. PMID- 8723923 TI - Single-step purification/solubilization of recombinant proteins: application to surfactant protein B. PMID- 8723924 TI - Quantitative dot-immunoblot assay to measure protein in multiple brain samples. PMID- 8723925 TI - Conversion of pipet-tip boxes into secure storage containers for microcentrifuge tubes. PMID- 8723926 TI - Screening for recombinant plasmids in yeast colonies of the two-hybrid system using PCR. PMID- 8723927 TI - Transient transfection of primary cultured hepatocytes using CaPO4/DNA precipitation. AB - We present a detailed protocol for the transient transfection of non proliferating primary cultured hepatocytes that is easily reproducible. Using a modification of the classical CaPO4/DNA precipitation method, this protocol is an inexpensive alternative to other methods that are often cumbersome, expensive, difficult to reproduce or harmful to primary hepatocytes. Because only 0.5 x 10(6) cells are needed for a single transfection experiment, several reporter genes can be introduced into hepatocytes of a single liver preparation. With our protocol, different plasmids can be introduced into one cell. In this way, cis trans interactions can be examined and reporter gene expression can be normalized for transfection efficiency. Furthermore, we describe details of a transfection experiment with two different reporter gene vectors using a luciferase gene and a lacZ gene. The results presented may be helpful to other groups concerned with improved timing of transfection experiments. PMID- 8723928 TI - In vivo chromatin structure of the murine interleukin-5 gene region: a new intact cell system. AB - The study of chromatin involvement in the regulation of gene expression has traditionally required the isolation of nuclei. However, cell fractionation techniques are subject to redistribution of proteins during the isolation procedure, which prevents rigorous physiologically relevant analysis. To eliminate the need to isolate nuclei and to analyze chromatin structures in vivo in response to agents regulating murine interleukin-5 (IL-5) gene activation, we have established a novel lysolecithin permeabilized intact cell system for suspension cell types, in this case T cells. Nuclear integrity of permeabilized cells is demonstrated by nuclear transport assays using confocal laser scanning microscopy. Results are identical in unstimulated and stimulated T cells, indicating that the chromatin structure after activation is not the result of gross alterations in nuclear protein transport properties. Potential new IL-5 gene regulatory regions are identified by DNase I hypersensitivity mapping. Our lysolecithin permeabilized intact cell system is amenable to physiologically relevant analysis of responses to signaling pathways at the level of chromatin, nuclear protein translocation and possibly other cellular functions in a variety of suspension and adherent cell types. PMID- 8723929 TI - Single-step immunocapture RT-PCR in the detection of raspberry bushy dwarf virus. AB - An immunocapture reverse transcription polymerase chain reaction (IC-RT-PCR) method for a highly sensitive analysis of raspberry bushy dwarf virus (RBDV) in infected plants is described. In the method, preliminary purification of virus particles or viral RNA from the plant material is not necessary. Viruses are enriched during the assay by antibodies bound in the PCR microplate wells, followed by lysis of the viral particles, and RT-PCR of the viral RNA. The reaction mixtures, including reverse transcriptase and DNA polymerase, have been selected so that both enzymes are active in the lysis and amplification conditions; by this way, it is possible to conduct the whole procedure in a single step. Using the method, four fragments from RNA-3 of RBDV have been amplified with various combinations of four primers. The procedure is sensitive enough to allow a simple detection of RBDV in in vitro cultured plants in which the detection of viruses by conventional immunological methods is difficult or even impossible. PMID- 8723930 TI - Single-tube protocol for the extraction of DNA or RNA from paraffin-embedded tissues using a starch-based adhesive. AB - We describe a method using an inexpensive craft glue to routinely isolate specific areas of tissue as small as 1 mm2 from paraffin sections. The tissue may be digested to release nucleic acid suitable for PCR or reverse transcription PCR. The use of this procedure obviates the requirement for manual microdissection or ultraviolet light irradiation. Tissue remaining on the slide can be stained and analyzed, allowing the precision of the extraction to be determined. The slide can be stored as a permanent record of the material taken for analysis. PMID- 8723931 TI - Improved method for the production of M13 phage and single-stranded DNA for DNA sequencing. AB - An improved method is described for the efficient production of M13 phage and M13 single-stranded (ss)DNA in a relatively short time period. Infection of E. coli (F') cells with as few as 5 phage particles can yield 10(12) phage particles/mL in 3 hours if the cells are grown in LB broth or SOB broth supplemented with about 5 mM Mg2+. The method tolerates large variations in the initial multiplicity of infection (5-5000 phage per 5 x 10(7) cells) and still yields about 10(12) phage particles/mL. These amounts are sufficient to purify 10-15 micrograms of ssDNA and to carry out at least 10-15 DNA sequencing reactions. PMID- 8723932 TI - Sequencing homopolymer tracts and repetitive elements. AB - We investigated the use of Taq dye primer and Taq terminator sequencing chemistry to optimize the quality of sequence data obtained from templates containing homopolymer tracts and repetitive elements. In direct side-by-side comparisons using the Applied Biosystems Model 373A Fluorescent Sequencer, the Taq terminator sequencing chemistry gave much cleaner and more consistent results on long homopolymer tracts and dinucleotide repeats. We also investigated various thermal cycling conditions and determined that higher annealing temperatures and longer denaturation times improved the ability to sequence through these problem templates. PMID- 8723934 TI - Prevention of radioactive contamination of cell-culture incubators during metabolic labeling experiments. PMID- 8723933 TI - A control system for cDNA enrichment reactions. AB - Most cDNA enrichment techniques involve technically complicated subtractive and hybridization reactions whose kinetics have not been characterized. We describe a control for cDNA enrichment procedures based on commercially available reagents, and we use this control to optimize a commonly used subtractive hybridization reaction. Using this control, we show that high-abundance transcripts can be efficiently removed following very short hybridization reactions. With the addition of unlabeled rat liver cDNA, we develop a system that mimics the reaction kinetics of complex cDNA pools. Because this system enables the measurement of specific vs. nonspecific hybridization and subtraction in the same tube and is an effective control for all steps in the cDNA enrichment procedure, it facilitates the development and optimization of novel cloning techniques for a desired abundance level of differentially expressed cDNAs. PMID- 8723935 TI - Economical apparatus for safe, accurate recovery of biohazardous or radioactive gradient fractions. PMID- 8723936 TI - Comments on a recently published PCR method for site-directed mutagenesis. PMID- 8723937 TI - Multiplex sets for the amplification of polymorphic short tandem repeat loci- silver stain and fluorescence detection. AB - Multiplex PCR amplification systems were developed using well-characterized, polymorphic short tandem repeat (STR) loci. Eight loci utilized in the multiplex amplifications included HUMCSF1PO, HUMTPOX, HUMTH01, HUMVWFA31, HUMF13A01, HUMFESFPS, HUMBFXIII and HUMLIPOL. From this list, three or four non-overlapping loci were simultaneously amplified, separated by denaturing polyacrylamide gel electrophoresis and visualized using silver stain or fluorescence detection. The multiplex PCR amplification systems offer a non-isotopic method for rapid, simple and accurate analysis of STR loci. This high-throughput method for DNA identification has immediate and valuable application in forensic analysis, paternity determination, tissue culture strain identification and bone marrow transplantation studies. PMID- 8723938 TI - Rapid, high-level transient expression of papillomavirus-like particles in insect cells. AB - Empirical scanning of natural or engineered peptide sequences for functional residues is inherently dependent upon efficient expression of large numbers of individual sequence variants to assay their relative functional potency. The insect baculovirus system has been widely used for expression of viral coat proteins, but it generally requires prior isolation and expansion of a plaque purified recombinant viral stock to generate useful quantities of self-assembled virus-like particles. In search of a more rapid means of expression of analytical levels of the L1 coat protein of cottontail rabbit and human type 11 papilloma viruses, we found that even brief transient cotransfection of insect cells with baculovirus plasmid transfer vectors and viral DNA yielded assembled particles that were immunologically indistinguishable from particles obtained with plaque purified viral stocks. Within six days of plasmid/viral DNA cotransfection of Sf9 cells, at least 1-2 micrograms of assembled L1 particles/100-mm plate could be demonstrated, which proved more than sufficient to assay functionality. Transient cotransfection of insect cells should provide general utility for rapid high level expression of sets of protein sequence variants, as well as other sequence scanning applications such as sequence optimization in protein engineering. PMID- 8723939 TI - CD-tagging: a new approach to gene and protein discovery and analysis. AB - We describe a new method for gene discovery and analysis, CD-tagging, that puts specific molecular tags on a gene, its transcript and its protein product. The method has been successfully tested in two organisms, the haploid unicellular alga Chlamydomonas reinhardtii and the metazoan Drosophila melanogaster. The method utilizes a specially designed DNA molecule, the CD-cassette, that contains splice acceptor and donor sites surrounding a short open reading frame. Insertion of the CD-cassette into an intron in a target gene introduces a new exon, represented by the open reading frame of the CD-cassette, surrounded by two functional hybrid introns. As a result (i) the gene is tagged by a specific nucleotide sequence, (ii) the mRNA is tagged by a specific nucleotide sequence and (iii) the protein is tagged by a specific peptide sequence. Because these tags are unique, specific nucleotide or antibody probes can be used to obtain and/or analyze the gene, transcript or protein. As a gene discovery technology, CD-tagging has two unique advantages: 1) Genes can be identified through a primary screen at the protein level, and so the very process by which a gene is identified provides specific empirical information about its biological function. 2) The cassette arms, which are spliced out of the transcript of the target gene, are available to carry a wide variety of DNA sequences, such as genes encoding drug resistance that can be used to select for the presence of the CD-cassette in the genome. PMID- 8723940 TI - Glycerol enhancement of ligand-polylysine/DNA transfection. AB - Primary human fibroblasts and a series of cell lines (A549, BNL CL.2, H225, NIH 3T3 and Rat-1) are efficiently transfected by using positively charged complexes of plasmid DNA and transferrin-polylysine or polylysine in the presence of glycerol (1 molar to 1.8 molar, depending on the cell type). An increase in gene expression of up to several-hundredfold (compared to complexes without glycerol) is obtained if the transfection mixture is incubated with the cells for 3-4 h at 37 degrees C. This simple method has been used for transient expression of luciferase, beta-galactosidase and interleukin-2, and also for the generation of stably transfected cells. PMID- 8723941 TI - Accelerating the pace of luciferase reporter gene assays. AB - Luciferase reporter gene assays have gained more importance because of their easy readout, high sensitivity and lack of environmental waste disposal problems. However, several obstacles remain that have prohibited a wider use and the implementation of this type of assay in high-throughput screening programs: (i) Measurements need to be carried out within an active enzyme reaction, and the assessment of such reactions are time-dependent; (ii) the signal produced has a "flash" type characteristic and therefore requires specialized equipment for measurement; and (iii) side-reactions can occur that interact with the signal readout of the assay in a non-reproducible way. These hurdles make an otherwise convenient assay principle troublesome for larger-scale screening use. We have attempted to overcome these problems by different means, leading to the development of LucLite, a stable signal homogeneous reagent system. This system allows use in a higher throughput screening capacity and enables the use of standard scintillation/luminescence instruments. PMID- 8723942 TI - Better gel resolution and longer cDNAs increase the precision of differential display. PMID- 8723943 TI - The contralateral ear in patients with chronic otitis media. AB - The status and hearing function of the contralateral ears in 493 patients undergoing surgery for chronic otitis media were studied at the final follow-up examination of the patients (on average, 6.5 years after the surgical treatment). In only 37% of the patients was the contralateral ear found to be normal, with atrophy of the pars tensa and tympanosclerosis being the most common abnormal findings, followed by pars tensa and attic retractions. In 18% of the patients, the contralateral ear had also undergone surgery. In only 64% of the patients, the contralateral ears had normal (< or = 20 dB) hearing levels, and in 21%, the hearing levels were > 30 dB, the hearing function of patients with cholesteatoma being worse than that of other patients with chronic otitis media. In 14 (3%) of the patients, even the better hearing ear had a hearing level of > 60 dB, including one patient with both ears totally deaf. To improve the functional outcome of patients with chronic otitis media, earlier and more effective treatment of middle ear infections is needed. PMID- 8723944 TI - Surgical anatomy of the epitympanic space. AB - A clear understanding of the surgical anatomy of the epitympanic space is crucial in modern otologic surgery. The tortuous facial nerve within the tight confines of this region presents important surgical challenges, especially when normal anatomy is obscured by disease. Microdissections of 20 human temporal bones were carried out with particular emphasis on the spatial relations of the facial nerve with various temporal bone landmarks. Measurements were made through a standard transmastoid approach to ensure clinical relevance. A high margin of safety can be achieved by observing consistent landmarks and relations in dealing with the facial nerve. We demonstrated the limitations of the transmastoid approach, particularly in accessing the labyrinthine segment of the facial nerve. PMID- 8723945 TI - A morphological study on obliteration of the temporal dorsal bullae using hydroxyapatite granules. AB - The present study deals with morphological observations of temporal dorsal bullae obliterated by implantation of hydroxyapatite granules. The specimens were obtained from guinea pigs 1 year after implantation, and undecalcified tissue sections were examined under a light microscope. The temporal dorsal bullae were found to be completely filled with bone tissue including the hydroxyapatite granules in all animals examined. The bone and hydroxyapatite granules interdigitated tightly without any intervening connective tissue or dead space. The newly formed bone displayed incomplete osteon structures with lamellae surrounding a haversian canal. There was no evidence of inflammation, such as lymphocyte infiltration, in the temporal bullae. The bone formation induced by hydroxyapatite implantation appeared to be a physiological reaction. It was concluded that the hydroxyapatite granules are safe and useful for mastoid obliteration during tympanoplasty. PMID- 8723946 TI - Ossiculoplasty by interposition of a minor columella between the tympanic membrane and stapes head. AB - This study analyzes short- and long-term results of 436 ossiculoplasties accomplished by a minor columella sculptured in the remnants of the incus and placed between the tympanic membrane and stapes head. Most cases were operated on by a transcanal approach through an ear speculum, under local anesthesia. Cases in which an associated mastoidectomy was performed to remove diseased tissue are excluded. The residual air-bone gap was < 20 dB in 86.1% of cases 2 months after surgery, in 77.0% 1 year later, and then remained stable over time, even in patients seen 5 to 15 years after the operation. Complications were rare. A total sensorineural hearing loss occurred in two cases (0.4%), the cause of which remained unexplained. Other complications included a high-frequency hearing loss in 10 (2.3%) cases, tinnitus in 12 (2.7%), and transitory vertigo in two (0.4%). We conclude that this type of ossiculoplasty is a simple but adequate procedure for primary restoration of hearing in chronic otitis media if the stapes is intact and mobile. PMID- 8723947 TI - Epidermal growth factor receptor (EGF-R) in human middle ear cholesteatoma: an analysis of protein production and gene expression. AB - Previous studies have shown an altered epithelial cell proliferation in middle ear cholesteatoma, reporting an aberrant expression of epidermal growth factor receptor (EGF-R) glycoprotein by immunohistochemistry. In this study, we quantified the presence of EGF-R using enzyme-linked immunosorbent assays (ELISAs) on tissue extracts, as well as the EGF-R gene expression by in situ hybridization on frozen sections. Human skin obtained from the external ear canal was used as control. The amounts of EGF-R glycoprotein in cholesteatoma were very similar to those in human skin. Human skin showed EGF-R messenger RNA (mRNA) only in the basal layer. A higher percentage of cells hybridized for the anti-sense probes EGF-R was found in cholesteatoma epithelium. Furthermore, we could find suprabasal cells with EGF-R mRNA. Our results confirm that the abnormal growth of cholesteatoma epithelium is reflected in an aberrant expression of EGF receptor. PMID- 8723948 TI - In vitro susceptibility of aural isolates of Pseudomonas aeruginosa to commonly used ototopical antibiotics. AB - The choice of antimicrobial agents used to treat Pseudomonas aeruginosa infections of the ear is quite empiric. Yet in spite of this, very little has been published examining susceptibility patterns of aural isolates of P. aeruginosa. Recently, increasing concern has emerged over the development of resistance to many of the commonly used ototopical preparations with activity against P. aeruginosa. This concern stems from the fact that these preparations have been in use for a long time, and P. aeruginosa is known to develop resistance fairly readily. We prospectively studied the susceptibilities of aural isolates of P. aeruginosa in 231 consecutive children who were seen in the outpatient Pediatric Otolaryngology Department at Children's Hospital of Pittsburgh during the years 1992 and 1993. The agents tested included neomycin, polymyxin B, colistin, and norfloxacin. We found that only 17.8% of the isolates were sensitive to neomycin, as opposed to > 95% for each of the other agents tested (polymyxin B, 99.6%; colistin, 97.4%; and norfloxacin, 98.3%). This difference proved to be statistically significant (p < 0.05). Given the concern of aminoglycoside-induced ototoxicity and the high rate of neomycin resistance, we believe that further investigation of other alternative ototopic agents with activity against P. aeruginosa is warranted. PMID- 8723949 TI - Stapedectomy in residency training. AB - Because fewer stapedectomies are performed today, concern has focused on whether or not residents are adequately trained in stapedectomy techniques. A retrospective study was done of patients undergoing stapedectomy over the last 11 years. This series represents primary resident-assisted cases performed under direct supervision of the attending surgeon. For 102 consecutive stapedectomies, the air-bone gap was closed to within 10 dB in 91 (89%), and no serious complications occurred. These results are compared with other published results, and our philosophy of teaching stapedectomy in residency is discussed. PMID- 8723950 TI - Reporting operative hearing results in stapes surgery: does choice of outcome measure make a difference? AB - In a prior study, findings indicated that when reporting results of chronic ear surgery, neither choice of pre-versus postoperative bone-conduction scores nor choice of frequencies to include a averaging makes a substantial difference in reported outcome. In this study, audiologic data from 240 stapes-surgery patients at three different institutions were used to generate a variety of outcome measures. Use of preoperative rather than postoperative bone-conduction values in computing postoperative air-bone gap resulted in an approximately 5-dB smaller mean gap and a 2% higher success rate. Frequencies included in averaging made little difference in mean computed air-bone gap, although success rate (gap < 10 dB) was lower by 6% when 4 kHz was used in a four-frequency average rather than 3 kHz. Results for air conduction were similar to those for air-bone gap regarding choice of frequencies to include in averaging. When using air-conduction pure tone average (PTA) as the outcome measure, those with normal preoperative sensorineural hearing had a > 20% higher success rate than the general population of stapes-surgery patients. The greatest differences in success rate were based on definition of and criteria for success. Success rate was higher when based on air-bone gap than when based on air-conduction PTA. As in the prior chronic ear study, differences in outcome were more drastically affected by criteria for "success" than by frequencies included. Unlike similar data from chronic ear surgery, however, success rate differed depending on choice of air-bone gap or air-conduction PTA as the definition for success. Further, air and bone scores from the same test interval must be used to accurately reflect air-bone gap in stapes surgery. PMID- 8723951 TI - A comparison of the results of KTP and CO2 laser stapedotomy. AB - Over the last 10 years, lasers have evolved to become an integral part of stapes surgery. Argon, KTP, and CO2 lasers have all been tested for their improved accuracy and safety in the middle ear. Because of the difference in optical properties between visible light (argon, KTP) and infrared (CO2) lasers, discussions have evolved around which laser is better and safer. In an effort to clarify the issues, a prospective comparison was made of 100 consecutive primary stapedotomies by using either the CO2 or KTP laser. PMID- 8723952 TI - Revision stapedectomy with and without the CO2 laser: an analysis of results. AB - A retrospective review is presented of revision stapedectomies performed by the senior authors between 1986 and 1994. A total of 106 stapedectomies was performed during that period, of which 30 were revisions. These revisions were within the first 100 stapedectomies performed by the senior authors. Most failures occurred early in the series. The overall success rate for closure to within 10 dB was 52%, and the average closure was 12.72 dB. Five cases were performed without the laser with no successes and an average closure of 27 dB. After the use of the laser, the success rate was 64%, and the average closure was 9.75 dB. There was a statistically significant difference between the hearing results with and without the laser (p < 0.01). The prognosis was better when surgery was performed primarily for hearing loss and with the laser, with success in 13 (72%) or 18 cases. In two cases, there was a sensorineural decline > 10 dB. The only dead ear occurred preoperatively in a patient with a granuloma. We have found the use of the CO2 laser in revision stapedectomy to be a safe technique that produces reliable results. PMID- 8723953 TI - Vein graft in stapes surgery. AB - Sealing the opening of the oval window during stapes surgery is essential; it prevents postoperative complications, such as perilymph fistula and sensorineural hearing loss. In this small series of 269 cases with otosclerosis, tympanosclerosis, and congenital ossicular abnormality, vein grafting was used to seal the opening of the footplate. Hearing improvement after surgery was acceptable, and none had total hearing loss or perilymphatic fistula. World literature from the last half of this century on grafting the oval window is reviewed. Absorbable gelatin sponge (Gelfoam) seems to be causing more complications, so its use is highly discouraged. Temporalis fascia, fat, and perivenous loose areolar tissue have been used by different authors at different times in footplate surgery. The opening created in the oval window during stapes surgery must not be left uncovered. PMID- 8723954 TI - Neurophysiological approach to tinnitus patients. AB - The principal postulate of the neurophysiological model of tinnitus is that all levels of the auditory pathways and several nonauditory systems play essential roles in each case of tinnitus, stressing the dominance of nonauditory systems in determining the level of tinnitus annoyance. Thus it has been proposed to treat tinnitus by inducing and facilitating habituation to the tinnitus signal. The goal is to reach the stage at which, although patients may perceive tinnitus as unchanged when they focus on it, they are otherwise not aware of tinnitus. Furthermore, even when perceived, tinnitus does not evoke annoyance. Habituation is achieved by directive counseling combined with low-level, broad-band noise generated by wearable generators, and environmental sounds, according to a specific protocol. For habituation to occur, it is imperative to avoid masking tinnitus by these sounds. Since 1991, > 500 tinnitus patients have been seen in our center. About 40% exhibited hyperacusis to varying degrees. A survey of > 100 patients revealed > 80% of significant improvement in groups of patients treated with the full protocol involving counseling and the use of noise generators. Notably, in patients who received counseling only, the success rate was < 20%. The improvement in hyperacusis was observed in approximately 90% of treated patients. PMID- 8723955 TI - Pulsatile tinnitus associated with congenital central nervous system malformations. AB - Pulsatile tinnitus as a manifestation of congenital central nervous system malformations has not been previously described. We present two patients with type I Arnold-Chiari malformation and one patient with congenital stenosis of the sylvian aqueduct with pulsatile tinnitus. Pulsatile tinnitus in these patients is believed to be secondary to increased intracranial pressure. Tinnitus subsided in one patient with Arnold-Chiari malformation after suboccipital decompression. The other two patients declined surgery. Audiologic and radiologic findings are presented, and our diagnostic algorithm for pulsatile tinnitus is described. The pathophysiology of pulsatile tinnitus can be a manifesting symptom in some patients with congenital central nervous system malformations. PMID- 8723956 TI - A human temporal bone study of changes in the basilar membrane of the apical turn in endolymphatic hydrops. AB - We observed that some temporal bones with endolymphatic hydrops (EH) showed varying degrees of basalward displacement (towards the scala tympani) of the basilar membrane (BM) in the apical turn of the cochlea. In some, the BM was adherent to the bony wall of the scala tympani (i.e., the interscalar septum). Such mechanical distortion of the BM could conceivably alter cochlear mechanics and lead to sensorineural hearing loss. The results of a systematic evaluation of 234 temporal bones to characterize, quantify and determine the functional significance of this observation are presented. Four groups of bones were evaluated: normal (N = 78), presbycusis (N = 96), Meniere's disease (N = 23), and EH secondary to labyrinthitis (N = 37). The incidence of extreme displacement of the BM in the apical turn such that it adhered to the interscalar septum was 52% in Meniere's disease, 57% in EH secondary to labyrinthitis, 10% in presbycusis, and 1% in normals. These differences were significant and could not be explained on the basis of age, sex, postmortem time, or artifact of technique or processing. Displacement of the BM was not observed in other turns of the cochlea. Its pathogenesis is not known, but may be related to atrophy of the spiral ligament. It is likely that such BM displacement results in sensorineural hearing loss. However, our data and theoretical analyses both indicate that such a loss will be restricted to frequencies below 100 Hz and that this pathologic change alone is not likely to cause appreciable hearing loss at clinically tested frequencies of 250 HZ and higher. Hence, even though this pathologic finding is common in endolymphatic hydrops, it cannot explain the low-frequency hearing lost observed in Meniere's disease. PMID- 8723957 TI - Vestibular neurectomy: retrosigmoid-intracanalicular versus retrolabyrinthine approach. AB - Selective vestibular neurectomy is an effective treatment for intractable vertigo of peripheral vestibular origin when preservation of hearing is a goal. The retrolabyrinthine and retrosigmoid-intracanalicular approaches have been used predominantly at our institutions over the last 10 years. The results and complications of these two techniques were compared. No significant differences were found between hearing results in these two patient groups. The retrosigmoid internal auditory can approach yielded better control of recurrent episodic vertigo, as well as superior ablation of postoperative ice-water caloric responses (p < 0.05). Surgical complications, including the incidence of cerebrospinal fluid leakage (greater in retrolabyrinthine approach) and postoperative headache (more prevalent in retrosigmoid approach), were also analyzed. To further evaluate the results of this study, data were reanalyzed and compared with previously published reports of selective vestibular nerve section. PMID- 8723958 TI - Fluorescein as an easy, low-cost, indirect, or reverse intraoperative marker to rule out perilymph versus local injection. AB - Clear fluid found in the dependent portions of the middle ear (round and oval windows) is generally assumed to be perilymph (PL). However, all clear fluid observed at middle ear exploration is not necessarily PL. PL should be distinguished from local injection. An accumulation of the local injection could be confused with perilymph fistula (PLF) diagnosis. There is no standard way of distinguishing perilymph from local injection, as both are clear, watery fluids. Fluorescein, tagged with a mixture of xylocaine and epinephrine (FLOTAX) was used as the local injection in 10 patients undergoing middle ear exploration for possible PLF.FLOTAX was injected into the vascular strip and ear canal skin in routine fashion. Transtympanic endoscopy and special Zeiss custom fluorescein filters were used to document any accumulation of fluid in the dependent portions of the middle ear and confirm whether any fluorescein (from the FLOTAX) was present. The middle ear was checked both before (endoscopically) and after raising the tympanomeatal flap. After the "local" injection, FLOTAX was observed to accumulate in dependent portions of the middle ear before raising the tympanomeatal flap in six of 10 ears. In the other four ears, FLOTAX slowly seeped into the middle ear cleft after the tympanomeatal flap was elevated and manipulated. Fluorescein can be used as an inexpensive, indirect intraoperative marker or "reverse test" for possible PLF. By eliminating a likely and common offender (local injection), any accumulation or reaccumulation of clear fluid that is not fluorescein tagged may more confidently be identified as possibly perilymph. The use of FLOTAX helps eliminate the confusion between the local injection and potential PLF. It does not in any way eliminate the possibility of other transudates from incisions, allergic reactions, or other manipulation of the tympanomeatal flap or middle ear mucosa. PMID- 8723959 TI - Vestibular symptoms in otosclerosis--correlation of otosclerotic involvement of vestibular apparatus and Scarpa's ganglion cell count. AB - Although several histopathologic studies have shown otosclerotic involvement of the vestibular apparatus in patients with otosclerosis, the pathogenesis of vestibular symptoms in otosclerosis remains unknown. A quantitative study of Scarpa's ganglion was performed in 217 temporal bones from 118 subjects with otosclerosis. Review of clinical records revealed an incidence of vestibular symptoms in 11.9% of these subjects. Scarpa's ganglion cell counts in temporal bones of subjects with otosclerosis and vestibular symptoms were lower than counts in temporal bones of subjects with otosclerosis but without vestibular symptoms and those of normal subjects. This difference in Scarpa's ganglion cell counts, adjusted for age, between the group with otosclerosis and vestibular symptoms and a group of normal subjects was highly significant (p = 0.0015), whereas the difference in Scarpa's ganglion cell count between a group with otosclerosis but without vestibular symptoms and a group of normal subjects was not significant (p = 0.53). There was also a significant correlation between elevation of the average bone-conduction threshold and the presence of vestibular symptoms in these subjects (p = 0.041). The endosteum of the perilymphatic space of the vestibule and the endosteum of the canal for the superior vestibular nerve or its cribrose area were the two most common sites of involvement by otosclerosis. However, there was no significant correlation between the presence of vestibular symptoms and otosclerotic involvement of any single site or the number of involved sites. Histologic examination of the vestibular nerve fibers and end organs subjacent to otosclerotic foci demonstrated no obvious degenerative changes. Thus our findings appear to suggest that the vestibular symptoms present in patients with otosclerosis are more common in patients with elevated bone conduction thresholds and are correlated with degeneration of the vestibular nerve, which appears to be independent of the severity of otosclerotic involvement of the vestibular end organs. PMID- 8723960 TI - Clinical utility of lateral head tilt posturography. AB - The Sensory Organization Test (SOT) of dynamic posturography, a functional test of balance, has a low sensitivity for compensated unilateral peripheral vestibular lesions. Tilting the head 45 degrees laterally during SOT [i.e. lateral heat tilt (LHT) SOT] may better detect such lesions. Normal subjects and vestibular clinic patients were studied using SOT and LHT SOT. Caloric, rotational chair, and oculomotor testing were also performed for patients, in either a double-blind or non-double-blind manner. The results for both patient groups were similar and thus were collapsed. Normative LHT SOT performance ranges were established to be slightly lower than corresponding SOT scores, for subjects aged 20-59 years. Subjects in older age groups performed with similar equilibrium scores in both SOT and LHT SOT. Normal subjects aged 20-59 and 60-69 years performed similarly, whereas those aged 70-79 performed significantly worse. Sensitivity of LHT SOT to unilateral peripheral vestibular lesions was poor (24%). Specificity of LHT SOT from patients having balance-system weakness was 90%. The results of this study indicate that LHT posturography does not facilitate significant discrimination of patients with compensated unilateral peripheral weakness from those without such lesions. PMID- 8723961 TI - The effect of endolymphatic sac excision in Meniere disease. AB - The role of the endolymphatic sac in the pathoetiology of Meniere's disease has been extensively studied but remains controversial. Likewise, the mechanism behind the efficacy of the endolymphatic-mastoid shunt procedure in Meniere's disease remains obscure. The central hypothesis of this study is that the endolymphatic-mastoid shunt procedure is a destructive rather than a drainage procedure. Therefore removal of the extraosseous endolymphatic sac with scarring of the intraosseous portion may be more destructive and thereby more effective in the treatment of Meniere's disease than conventional shunt placement alone in the extraosseous endolymphatic sac. This study reports a 2-year follow-up on the effect of complete removal of the extraosseous endolymphatic sac for 10 patients with intractable Meniere's disease and 10 matched control Meniere's disease patients undergoing conventional endolymphatic-mastoid shunt surgery. This study showed no statistical difference in the change in hearing, vertigo, or disability in the two groups. PMID- 8723962 TI - Craniometaphyseal dysplasia: operative findings and treatment. AB - Craniometaphyseal dysplasia (CMD) is a genetic bone disorder involving an abnormality of modeling of the long bones and sclerosis of the cranium. Both conductive and sensorineural hearing may be frequently associated with this syndrome. Conductive hearing loss is due to attic fixation of the lateral ossicular chain and hyperostosis formation with stapes ankylosis. Two patients with CMD were treated surgically. Two of three ears operated on had closure of the air-bone gap, whereas the third ear failed ossicular reconstruction because of hyperostosis of the promontory with deepening of the oval window. PMID- 8723963 TI - Congenital cholesteatomas of the middle ear: a different experience. AB - A retrospective analysis is presented of 44 cases of congenital cholesteatoma of the middle ear. Twenty-one patients had cholesteatoma localized to the posterosuperior mesotympanum in complete contrast to the commonly reported anterosuperior location seen in only two cases. The remaining 21 patients had cholesteatoma involving either the entire mesotympanum or epitympanum or both. The posteriorly located congenital cholesteatomas might represent a completely different entity and originate from the trapped epithelial cell rests in the posterior mesotympanum during the development of temporal bone. All but one patient were treated by using a closed tympanoplasty. Eight patients underwent a single-stage surgery. A preplanned second-stage procedure was performed in 33 patients, and three are awaiting their second stage. Residual disease was seen in 19 (57%) patients undergoing second-stage surgery. No patient has had recurrent disease so far. Thirty-eight (85%) patients had a preoperative air-bone gap of > or = 30 dB. Of the 33 patients considered for hearing results, 16 (48%) had a postoperative gap within 10 db. PMID- 8723964 TI - Type I tympanoplasty in children. AB - To identify factors that could influence the success of tympanoplasties in children, a retrospective study was done of 51 tympanoplasties performed on children aged 9-14 years during the years 1984-1989. Surgical success was defined as an intact membrane found after 3 years' follow-up. The success rate of tympanoplasties was 92%. A functioning eustachian tube was found to be important for graft take. No difference was found in the rate of graft take whether fascia of the temporal muscle or tragal perichondrium was used. An audiologic improvement was seen in 70.6% of the patients. We conclude that tympanoplasty in children has a high probability of success and will enable most children operated on to resume a normal lifestyle. PMID- 8723965 TI - A unique temporal bone lesion resembling juvenile active ossifying myxoma. AB - Juvenile active ossifying fibroma (JAOF) is a lesion characterized by early age of onset, slow growth, frequent recurrence, and locally aggressive behavior. Histologically, it consists of three major components: cellular fibrous stroma, osteoid bodies, and myxomatous matrix, which may become cystic. This article describes a case of a slowly growing destructive lesion of the mastoid in a 2 year-old girl with histologic features resembling those of juvenile active ossifying myxoma (JAOM). Histologically, its prominent features were myxoid and fibromyxoid stroma with cystic areas, vascular spaces, bone, and multinucleated giant cells. This is the first pediatric temporal bone lesion with these features recorded These histologic components led to the diagnosis of JAOM of the temporal bone, probably developing in relation to the development of the mastoid air spaces. PMID- 8723966 TI - Growth of vestibular schwannomas: in situ model employing the monoclonal antibody Ki-67 and DNA flow cytometry. AB - Vestibular schwannoma (VS) growth potentials were studied in an in situ model, in which the cycling cellular fraction was determined immunohistochemically by applying the mouse monoclonal Ki-67 antibody, and the tumor ploidy was estimated by DNA flow cytometry in a consecutive series of 124 VSs. The tumors were classified according to the average number of positively stained nuclei in 10 high-power fields into three groups: 28 highly (> 10), 33 moderately (> 5-10) and 63 low proliferating (< or = 5). The intratumoral proliferative variation was studied in 10 tumors. Only slight variation in the number of the positively stained nuclei were observed. Six of seven tumors removed because of macroscopically documented growth by computed tomography (CAT) scan were moderately or highly proliferative. Proliferation of VS was correlated to prospectively registered clinical data. A statistically significant relation was found between VS proliferation and the prediagnostic duration of symptoms (p = 0.0001). The proliferative status was unrelated to age, sex, and tumor size. Flow cytometric determination of DNA index of the 124 tumors revealed 12 tetraploid (DNA index = 2), 110 diploid (DNA) index = 1) and two nondiploid tumors. A statistically significant relation was noted between tumor ploidy and proliferation status expressed by Ki-67 (p = 0.024). The tetraploid tumors showed significantly lower proliferation compared with the diploid tumors. Tumor ploidy was statistically unrelated to age, sex, tumor size, and duration of symptoms. The results of this study provide a link between the immunohistochemical, flow cytometric findings, and clinical data, which could probably be relevant in identifying patients at risk for rapid tumor growth and tumor recurrences, because a rapid test for cell proliferation is now available. PMID- 8723967 TI - Acoustic neuroma recurrence after suboccipital resection: management with translabyrinthine resection. AB - Thirty-five patients with recurrent acoustic tumors have been studied before, during, and after revision surgery. Each had their original surgery via a suboccipital approach. Six patients had undergone multiple suboccipital surgeries. Revision surgery was accomplished using a translabyrinthine approach in all patients. Total tumor removal was possible in 33 patients. No second recurrences have occurred to date among the patients with total tumor removal. Anatomic facial nerve integrity was maintained in 24 patients. All recurrent tumors were present in the lateral internal auditory canal. Direct tumor access in this area is not possible with the suboccipital approach without entering the vestibular labyrinth. The suboccipital approach to acoustic neuromas is contraindicated if tumor extends to the fundus of the internal auditory canal. Translabyrinthine resection is the treatment of choice for recurrence of acoustic neuroma after primary suboccipital resection. PMID- 8723968 TI - Correlation of hearing loss and radiologic dimensions of vestibular schwannomas (acoustic Neuromas). AB - A retrospective analysis was performed of puretone audiograms, speech discrimination scores, and gadolinium-enhanced magnetic resonance imaging scans of 75 patients with vestibular schwannomas (acoustic neuroma). Sensorineural hearing loss was analyzed for low frequencies (250-500 Hz), midfrequencies (1,000 2,000 Hz), and high frequencies (4,000-8,000 Hz). The largest tumor diameter in the cerebellopontine angle and the lateral extent of invasion by tumor into the internal auditory canal were calculated from magnetic resonance images. There were statistically significant correlations between the largest tumor diameter and the severity of low-frequency sensorineural hearing loss (p = 0.001). However, no significant correlations were found between the following: largest tumor diameter and the severity of mid-frequency or high-frequency sensorineural hearing loss or speech-discrimination scores and lateral extent of invasion of the internal auditory canal and sensorineural hearing loss at all frequencies or speech-discrimination scores (p > or = 0.05). The findings suggest that nerve compression is not the only cause of hearing loss in vestibular schwannoma. PMID- 8723969 TI - A prospective study of ABR and MRI in the screening for vestibular schwannomas. AB - The advent of magnetic resonance imaging (MRI) has significantly increased the clinician's ability to detect small vestibular schwannomas. This had led to controversy in the evaluation of patients with asymmetric sensorineural hearing loss, as some recent studies have suggested that the auditory brainstem response (ABR) does not adequately detect small tumors of the internal auditory canal and cerebellopontine angle. As these studies evaluated ABR results in patients already diagnosed with vestibular schwannomas, they could not determine the epidemiologic accuracy (validity) of the ABR as a screening test for retrocochlear pathology. We report on the preliminary results of an ongoing prospective study on the evaluation of patients with asymmetric sensorineural hearing loss. All patients with asymmetry in two or more pure-tone thresholds of > or = 15 decibels or asymmetry in speech discrimination scores of > or 15% or both entered the study and underwent both an ABR examination and an enhanced MRI scan. Based on preliminary results obtained from the first 47 patients entered in this study, the ABR screening test for retrocochlear pathology was determined to have a sensitivity of 63%, a specificity of 64%, a positive predictive value of 26%, and a negative predictive value of 89%. All patients in whom ABR failed to diagnose a vestibular schwannoma had unilateral hearing los. These results bring into question the validity of ABR as a screening test for retrocochlear pathology, particularly in cases of unilateral hearing loss. Continued patient enrollment in this study will allow the confirmation of these results. PMID- 8723970 TI - Delays in the diagnosis of acoustic neuromas. AB - From the medical files of 164 consecutive patients who underwent surgical treatment for a unilateral acoustic neuroma between 1980 and 1992, we collected data on the delay until the diagnosis was made. A distinction was made between the patient's and general practitioner's delay (delay 1) and the delay after the specialist's first visit until the radiologic diagnosis (delay 2). The average delay was 35.7 months (SD, 62.2) for delay 1 and 15.2 months (SD, 36.3) for delay 2. Specialist's delay (otolaryngologist or neurologist) was divided into a delay of a maximum of 12 months (134 patients) and a longer delay (30 patients). In 27 of the 30 patients, no specific tests had been performed, and in the remaining three, the test results were inconclusive. Reasons for not conducting further tests included familial hearing impairment, Meniere's disease, otosclerosis, and alcoholism. In cases in which the specialist had not made the diagnosis within 1 year, it took an average of 6 months extra to make the diagnosis of an acoustic neuroma, usually with a fairly short patient delay. The specialist's delay remained constant in the period of investigation, with the possibility of magnetic resonance imaging (MRI) scanning only in the last 2 years. In view of the increasing accessibility of MRI, it is now recommended if possible to perform MRIs in all patients with symptoms suspicious for an acoustic neuroma. PMID- 8723971 TI - Histopathology of the peripheral vestibular system in small vestibular schwannomas. AB - Gadolinium-enhanced magnetic resonance imaging can be used to detect small vestibular schwannomas/acoustic neuromas. Early detection raises the question of the necessity of their surgical removal. Do all tumors induce lesions in the vestibule and to what extent? We thus investigated the ultrastructure of peripheral vestibular systems in grades I and II schwannomas. Vestibular tissues were fixed as soon as they were removed during the resection of tumors, by the translabyrinthine approach, and then processed for transmission electron microscope observations. In neurosensory epithelia, hair cells lost stereocilia, whereas cuticular plates disaggregated. The cytoplasm of hair cells degenerated in either a dense or vacuolated manner, and cytoplasmic blisters extended into the endolymph. In some cases, supporting cells extended processes covering the apical surface of hair cells. Nerve fibers massively disappeared from epithelia, only few nerve fibers contacted type I and type II hair cells, and both afferent and efferent terminals were abnormal. In vestibular nerves, axons degenerated, and myelin sheaths disaggregated. Glycogen was present in both intracellular and extracellular spaces. Luse bodies associated with collagen bundles were found between fibers. Scarpa ganglion neurons contained lysosomes/lipofuscin granules and vacuoles. Tumor cells were found in both the ganglion and the vestibular nerve. Thus small tumors induce extensive degeneration of vestibular tissues. The various hallmarks of schwannomas are already present in small acoustic neuromas. Moreover, different types of degeneration of hair cells and neurons were observed, together with the covering of hair cells by supporting cells and the accumulation of glycogen in the vestibular nerve. PMID- 8723972 TI - Tubal and tympanic openings of the peritubal cells: implications for cerebrospinal fluid otorhinorrhea. AB - Cerebrospinal fluid otorhinorrhea after surgery for cerebellopontine angle tumors may persist despite obliteration of the mastoid, middle ear, and tympanic orifice of the eustachian tube. In this study, histologic sections of 120 adult temporal bones were examined by light microscopy to determine the incidence of peritubal pneumatization and to demonstrate the frequency of tubal and tympanic openings of the peritubal cells. The results of this study suggest that the pathway for these persistent cerebrospinal fluid leaks may be via the peritubal cells that open directly into the eustachian tube anterior to its tympanic orifice. Peritubal pneumatization was present in 78 (65%) of the temporal bones. Of the 57 specimens in which the openings of the peritubal cells could be identified, in 52 (91%), the cells opened into the eustachian tube anterior to its tympanic orifice, and in only five (9%), they opened into the middle ear. The overall incidence of tubal openings in this study was 59%. In 13 temporal bones (21%), the tubal opening were at a distance of > 5 mm anterior to the tympanic orifice of the eustachian tube. Therefore, cerebrospinal leak may persist through these tubal openings despite obliteration of the mastoid, middle ear, and tympanic orifice of the eustachian tube. A case of persistent cerebrospinal fluid leak in which extensive peritubal pneumatization was demonstrated by computed tomography scan is presented. Successful control of the leak was obtained only after the tubal openings of these cells several millimeters anterior to the tympanic orifice were obliterated. PMID- 8723973 TI - Defining functional limitation, disability, and societal limitations in patients with facial paresis: initial pilot questionnaire. AB - Experiences with patients with facial paralysis over the last 25 years and recent efforts to develop objective measures of paresis and synkinesis led us to three hypotheses: (a) dysfunction in certain regions of the face is more disturbing than that in others, (b) there are major psychosocial impacts of facial paralysis, and (c) the impact of facial paralysis is underestimated. An initial questionnaire of 10 open-ended items was submitted to 11 subjects stabilized after acoustic tumor resection. Responses were tabulated qualitatively, and frequency counts were made of responses. These results show that the region of the face that is most disturbing is the mouth; however, early in the time course of paralysis, the eye is most disturbing. Synkinesis above the eye is ultimately more disturbing than paresis in that region, and it may worsen. Major psychosocial impacts of paralysis appear common and underestimated. These pilot data qualitatively support the hypotheses. PMID- 8723974 TI - In vivo effects of surgical sympathectomy on intramembranous bone resorption. AB - Bone modeling and remodeling are highly regulated processes in the mammalian skeleton. The exact mechanism by which bone can be modeled at a local site with little or no effect at adjacent anatomic sites is unknown. Disruption of the control of modeling within the temporal bone may lead to various bone disease such as otosclerosis, osteogenesis imperfecta, Paget's disease of bone, fibrous dysplasia, or the erosion of bone associated with chronic otitis media. One possible mechanism for such delicate control may be related to the ubiquitous and rich sympathetic innervation of all periosteal surfaces. Previous studies have indicated that regional sympathectomy leads to qualitative alterations in localized bone modeling and remodeling. In this study, unilateral cervical sympathectomy resulted in significant increases in osteoclast surface and osteoclast number within the ipsilateral bulla of experimental animals. The mechanisms by which sympathectomy leads to increased local bone loss is unknown. Potential mechanisms include disinhibition of resorption, secondary to the elimination of periosteal sympathetics, as well as indirect vascular effects. PMID- 8723975 TI - Reconstructions and cross-sectional area measurements from magnetic resonance microscopic images of the cochlea. AB - In this study, magnetic resonance (MR) microscopy was used to obtain serial sections through the cochleae of mustached bats. As previously reported, 25 microns isotropic voxels can be obtained. Specific areas in each slice were segmented and then three-dimensional (3-D) reconstructions of the perilymphatic and endolymphatic spaces and spiral ligament were obtained. Quantitative measurements of the cross-sectional areas were made with customized macros written for the public-domain software, NIH Image. Results of this study revealed enlargements of the scalae and spiral ligament in areas known to be involved with processing of the animal's biosonar and fine-frequency analysis. PMID- 8723976 TI - Penicillin reduces secretory capacity in rat middle ear mucosa in acute otitis media. AB - In the United States, antibiotic treatment of acute otitis media is almost mandatory, whereas several other western countries are more reticent. Most clinical trails on antibiotic effect have important methodologic flaws, making an overall interpretation quite difficult. This study determined the effect of penicillin V administration on the secretory capacity of rat middle ear mucosa, during and after acute pneumococcal otitis media, by quantitative studies of the goblet cell density. The right middle ear bullae of 25 rats were inoculated with type 3 pneumococci. Beginning 2 days after inoculation, penicillin V 100 mg/kg/day was administered orally for 5 days. After inoculation, five randomly selected rats were killed on days 4, 8, 16, 90, and 180. The middle ear bullae were removed, split in half, stained with periodic acid-Schiff (PAS)-alcian blue, and the mucosae dissected from the bone. Whole mounts were prepared and the goblet cell density determined in 24 well-defined localities, making a total of 160 counts per ear and covering the entire bulla mucosa. Goblet cell densities were compared with those of 25 normal ears and 25 inoculated, untreated ears. Except on day 4, the penicillin V-treated ears had a significantly lower goblet cell density in almost all localities, on all days of death, when compared with untreated ears. Six months after the acute incident, the goblet cell density was almost normal. However, the enlargement of the mucosal area containing goblet cells seen in untreated ears was unaffected by penicillin V administration. We conclude that administration of penicillin V reduces the increase in secretory capacity of rat middle ear mucosa during and 6 months after acute pneumococcal otitis media. PMID- 8723977 TI - External and middle ear pathology in TGF-alpha-deficient animals. AB - Transforming growth factor-alpha (TGF-alpha) is a growth-regulatory peptide found in a wide range of embryonic and adult tissues. TGF-alpha is produced by keratinocytes and has been reported to be overexpressed in several epidermal diseases, including middle ear cholesteatoma. This report describes ear pathology in the waved-1 mutant mouse, which is severely deficient in TGF-alpha. Morphologic changes of the external and middle ear were studied histologically in waved-1 mutants 2 weeks to 6.5 months of age. Abnormalities found in the mutants included epidermal hyperplasia of the external ear canal (EAC) and tympanic membrane (TM) and enlargement of specialized sebaceous glands adjacent to the cartilaginous EAC. Sebum and desquamated keratin progressively accumulated within the EAC, displacing the TM into the middle ear. These changes appear similar to those occurring in Mongolian gerbils, which are known to develop cholesteatoma. The alterations found in waved-1 mutants are discussed in relation to the possible involvement of TGF-alpha in cholesteatoma pathogenesis. PMID- 8723978 TI - Antihypertensive therapy: safety and efficacy of drugs and publications. PMID- 8723979 TI - Genes for ischaemic stroke: strategies for their detection. PMID- 8723980 TI - Genotypes of sarco(endo)plasmic reticulum Ca(2+)-dependent ATPase II gene in substrains of spontaneously hypertensive rats. AB - OBJECTIVE: To search for a genetic marker of the sarco(endo)plasmic reticulum Ca(2+)-dependent ATPase (SERCA) II gene in spontaneously hypertensive rats (SHRs) and to investigate differences in blood pressure and intracellular Ca2+ among some substrains of SHRs and Wistar-Kyoto (WKY) rats related to their SERCA II genotypes. DESIGN AND METHODS: The coding region of the SERCA II gene was sequenced in SHRs. Blood pressure and intracellular Ca2+ concentration ([Ca2+]i) in platelets were measured in substrains of SHRs and WKY rats with different SERCA II genotypes. RESULTS: A point mutation that provided restriction fragment length polymorphisms (RFLPs) by HindIII or Saul was found in the SERCA II gene. The polymerase chain reaction (PCR) products were digested by HindIII in SHR substrains and WKY-Kyoto rats, whereas they were digested by Saul in normotensive strains and SHR-Toho. Among SHR-Kyoto, SHR-Toho, WKY-Kyoto and WKY-Charles River, the substrains with the HindIII-digested SERCA II genotype showed slightly but significantly higher systolic blood pressure and augmented agonist-stimulated [Ca2+]i than those with the Saul-digested genotype. CONCLUSIONS: RFLPs were found in the SERCA II gene. In the substrain analysis of SHRs and WKY rats, higher blood pressure and increased [Ca2+]i were associated with the SERCA II genotype digested by HindIII. The SERCA II gene locus has the potential to contribute to the development of hypertension and abnormal intracellular Ca2+ metabolism in SHRs. These RFLPs in the SERCA II gene should be a useful genetic marker. PMID- 8723981 TI - Left ventricular weight but not blood pressure is associated with sex chromosomes in Lyon rats. AB - OBJECTIVE: To investigate, by studying reciprocal back-crosses to Lyon hypertensive (LH) rats, the involvement of sex chromosome loci in high blood pressure and heart weight in LH rats. METHODS: Reciprocal F1 and F'1 generation male rats obtained from male LH x female Lyon normotensive (LN) and male LN x female LH crosses respectively, male back-cross rats obtained from male LH x female F1 or F'1 and from male F1 or F'1 x female LH crosses, and parental LH and LN male rats were studied at 29-31 weeks of age. Systolic, diastolic, mean and pulse pressures were measured beat to beat in unrestrained rats for 1 h. In addition, relative left and right ventricular weights were measured. RESULTS: The average blood pressures did not differ between F1 and F'1 rats or between the four populations of back-cross rats. On the contrary, the relative left ventricular weight was higher in F'1 than in F1 rats. As a role for loci on the Y chromosome could be discarded by comparison of the two populations of back-cross rats, which differ only in the origin of their Y chromosome, this increase in the relative left ventricular weight of F'1 rats was consistent with an effect of a locus on the LH X chromosome. This was supported by findings in the back-cross populations, those populations in which 100% of the rats carried an LH X chromosome having a significantly higher left ventricular weight than those in which only 50% of the rats carried such a chromosome. The estimate of the genetic determination was higher for left ventricular weight (41%) than for mean arterial pressure (19%). Although these two parameters were associated in the back-cross population, the origin of the X chromosome had no influence on this association. CONCLUSIONS: These findings indicate that, against the genetic background provided by the cross studied and at the age studied, the sex chromosomes of LH and LN rats do not contain loci at which alleles differentially influence blood pressure. They do, however, suggest that relative left ventricular weight in this model has a specific genetic determination, partly contributed by a locus on the X chromosome. PMID- 8723982 TI - Influence of insulin, sympathetic nervous system activity, and obesity on blood pressure: the Normative Aging Study. AB - OBJECTIVE: To examine the association of insulin and sympathetic nervous system activity with blood pressure elevation in a cross-sectional study of 752 nondiabetic male participants of the Normative Aging Study, aged 43-90 years. METHODS: Testing included a physical examination, medical history, fasting and post-carbohydrate insulin and glucose levels determinations, an anthropometric examination, and 24 h urine collection for catecholamine level determination. Total obesity was represented by body mass index, central obesity by the abdomen circumference:hip circumference ratio, and sympathetic nervous system activity by 24 h urinary excretion of norepinephrine. RESULTS: Systolic and diastolic blood pressure (SBP and DBP, respectively) were positively related to body mass index, abdomen:hip ratio, norepinephrine excretion, and insulin levels in univariate analyses. The relationship between insulin level and SBP and DBP persisted after adjustment for body mass index, abdomen:hip ratio, norepinephrine, age, smoking, physical activity level, and antihypertensive medication use. The norepinephrine level was related to SBP and DBP after adjustment for insulin level, age, smoking, physical activity level, and antihypertensive medication use, and these relationships remained marginally significant after further adjustment for body mass index and abdomen:hip ratio. In contrast, neither body mass index nor abdomen:hip ratio were related to blood pressure after adjustment for insulin level. Among participants in the lowest tertiles both of insulin and of norepinephrine levels, 10% were hypertensive, compared with 35% in the highest tertiles of these variables. In a multiple logistic regression model, insulin level, norepinephrine level, and an interaction term for insulin level with norepinephrine excretion were independent predictors of hypertension. CONCLUSIONS: The results suggest that insulin level and sympathetic nervous system activity are associated with hypertension among middle-aged and elderly men. PMID- 8723983 TI - Mortality and morbidity during a period of 2 years after coronary artery bypass surgery in patients with and without a history of hypertension. AB - OBJECTIVE: To describe mortality and morbidity during a period of 2 years after coronary artery bypass grafting (CABG) in relation to a history of hypertension. PATIENTS: All patients in western Sweden in whom CABG was undertaken between June 1988 and June 1991 and in whom simultaneous valve surgery was not performed were included in the study. DESIGN: A prospective 2-year follow-up study. RESULTS: Patients with a history of hypertension (n = 777) differed from patients without such a history (n = 1348) in that the proportion of women was higher, they were older and more frequently had a history of congestive heart failure, diabetes mellitus, renal dysfunction, cerebro-vascular disease, intermittent claudication and obesity, and the number of smokers and patients with previous CABG was lower. They were also more likely to develop post-operative cerebrovascular complications and signs of myocardia damage. Patients with hypertension tended to have increased mortality during the first 30 days after CABG and the late mortality (between day 30 and 2 years) was significantly higher than in non hypertensive participants. Whereas the development of myocardial infarction was similar in both groups, the hypertensive study participants more frequently developed stroke during 2 years of follow-up. In a multivariate analysis including age, sex, history of different cardiovascular diseases, smoking, ejection fraction, and the occurrence of three-vessel disease, hypertension did not emerge as an independent predictor of death in the early or late phase or during a total of 2 years of follow-up. CONCLUSION: Among CABG patients, those with a history of hypertension have a different pattern of risk factors. They have a higher mean age, include a higher proportion of women and have a higher prevalence of congestive heart failure, diabetes mellitus, renal dysfunction, cerebro-vascular disease, intermittent claudication, and obesity. They also have an increased frequency of immediate post-operative complications and an increased 2-year mortality, even if a history of hypertension was not an independent predictor of death during 2 years of follow-up. PMID- 8723984 TI - Angiotensinogen and blood pressure among blacks: findings from a community survey in Jamaica. AB - OBJECTIVE: To examine the association between blood pressure, angiotensinogen levels, angiotensin converting enzyme activity and polymorphisms of the angiotensinogen and angiotensin converting enzyme genes in a population-based sample. METHOD: Five hundred participants were recruited in a house-to-house survey of three communities in metropolitan areas of Kingston and St Andrew, in Jamaica. Demographic data, anthropometric and blood pressure measurements were obtained for each participant during a brief clinic visit. Circulating levels of angiotensinogen and angiotensin converting enzyme activity were measured in venous blood samples. Polymorphisms of the angiotensinogen and angiotensin converting enzyme genes were determined. RESULTS: A weak association between angiotensinogen level, angiotensin converting enzyme activity and blood pressure was identified in this population, but substantial joint effect of angiotensin converting enzyme activity and angiotensinogen level on blood pressure was apparent. Variants of the angiotensinogen gene had inconsistent effects on blood pressure and on the risk of hypertension. Angiotensinogen level and angiotensin converting enzyme activity were significantly related to several measures of obesity, including body mass index, waist circumference and skin fold thickness. CONCLUSION: The angiotensinogen and angiotensin converting enzyme genetic variants which were studied appear to have only a modest relationship with blood pressure and associated anthropometric risk factors among blacks. PMID- 8723985 TI - Patient assessment of self-measurement of blood pressure: results of a telephone survey in Germany. AB - OBJECTIVE: To obtain information on the adoption of self-measurement of blood pressure by hypertensives and their attending physicians. METHODS: Callers to the Cardiovascular Hotline Heidelberg revealing themselves to have high blood pressure were interviewed about self-recording of blood pressure. RESULTS: Among the callers about 70% practised self-measurement. Half of them bought a device on their own initiative, one-third on recommendation or prescription by their physician. A quarter of those practising the technique were introduced to it by experienced personnel and a comparable number were adequately informed about the aims and objectives. Nevertheless, more than half of those practising self measurement claimed to be coping better with the disease. CONCLUSIONS: The data indicate that self-monitoring is widely practised by hypertensives in Germany. In contrast to this finding, too few physicians have adopted home-recording as a useful tool in the treatment of hypertension. PMID- 8723986 TI - Isolated office hypertension: a prehypertensive state? AB - OBJECTIVE AND METHOD: Isolated office hypertension, defined as hypertensive blood pressure values in a medical setting but normal self-measured or ambulatory recorded blood pressures, is frequently encountered in clinical practice. Yet, whether this condition represents a transient state in the development of a sustained ambulatory hypertension is still unknown as no long-term analysis of the evolution of ambulatory blood pressure has been carried out in patients with isolated office hypertension. To evaluate whether such patients should be considered as truly normotensive or hypertensive, we have studied the long-term changes in office and ambulatory blood pressures in 81 patients in whom isolated office hypertension was observed between 1982 and 1988. RESULTS: After a 5-6 year follow-up, 60 of the 81 patients had a mean 12 h daytime ambulatory blood pressure greater than 140/90 mmHg, suggesting an evolution towards ambulatory hypertension. The development of hypertension could not be predicted on the basis of the follow-up office blood pressures as these tended to decrease during the follow-up period. CONCLUSIONS: The results of this study suggest that patients with isolated office hypertension should not be considered as truly normotensive individuals. Hence, these patients require a careful medical follow-up. Office blood pressure readings alone, however, do not appear to provide a good indicator of the long-term outcome of isolated office hypertension. PMID- 8723987 TI - Blood pressure rhythm and endocrine functions in normotensive women on oral contraceptives. AB - DESIGN AND METHODS: Twenty-four hour blood pressure profiles were determined by non-invasive ambulatory blood pressure measurements in young normotensive women during the follicular and luteal phases of the menstrual cycle. Forty women participated, 20 of whom were on oral contraceptives (ethinyl-ethylestradiol 0.03 mg + levonorgestrel 0.15 mg) and 20 of whom were age- and weight-matched control individuals not on oral contraceptives. Data on systolic and diastolic blood pressure and heart rate were analyzed in each case by linear and rhythm analysis. Urine was collected day and night on each occasion on which ambulatory blood pressure measurements were recorded. RESULTS AND CONCLUSION: Daytime, night-time, and 24 h mean systolic and diastolic blood pressure and heart rate did not depend on the cycle phase. The nocturnal fall in blood pressure was preserved in both groups and during both phases. Rhythm analysis by partial Fourier series showed that, of 240 individual 24 h blood pressure profiles, only 12 (5%) did not exhibit a significant circadian rhythm. Linear and rhythm analyses revealed that during both phases of the menstrual cycle systolic and diastolic blood pressure were significantly higher throughout the 24 h, especially during the night, in women taking oral contraceptives. During both phases of the menstrual cycle urinary aldosterone excretion was significantly higher in women taking oral contraceptives. This increase could contribute to the night-time blood pressure elevations caused by oral contraceptives. The results suggest a modulating influence of sex steroids on the circadian blood pressure profile even in normotensive healthy volunteers. PMID- 8723988 TI - Effect of angiotension II on the expression of the early growth response gene c fos and DNA synthesis in human vascular smooth muscle cells. AB - OBJECTIVES: The aims of this study were to characterize the angiotensin II receptor subtype present on vascular smooth muscle cells from human saphenous vein and to assess the effect of angiotensin II on the expression of the early growth response gene c-fos and on DNA synthesis. METHODS AND RESULTS: Using radioligand binding studies, we have defined the angiotensin II receptors present on these cells as being predominantly of the AT1 subtype. Angiotensin II increased peak intracellular calcium levels by 126 +/- 16 nmol/l (mean +/- SEM) in 17/49 cultures. Angiotensin II induced c-fos expression in a concentration dependent manner only in cultures that exhibited an intracellular calcium transient in response to stimulation with angiotensin II. The induction of c-fos was inhibited by the selective AT1 antagonist losartan in accordance with the binding studies. Angiotensin II stimulated DNA synthesis with a maximal increase of 66.4% +/- 20.5% over serum-free levels at 1 nmol/l (mean +/- SEM, n = 6, P < 0.05). DNA synthesis declined with increasing angiotensin II concentration, falling to control values at 1 mumol/l, suggesting that a growth-inhibitory influence may counter-balance the stimulatory effect that is observed at lower concentrations. CONCLUSION: Vascular smooth muscle cells from human saphenous vein possess predominantly AT1 receptors and in response to angiotensin II show an induction of c-fos and a modest increase in DNA synthesis. PMID- 8723989 TI - Angiotensin II receptor gene expression in hypertrophied left ventricles of rat hearts. AB - OBJECTIVE: To examine the expression of angiotensin II AT1a, AT1b and AT2 receptor genes in the left ventricles of rats subjected to ventricular pressure overloading induced by aortic banding for 6 weeks and then 6 weeks medical treatment. RESULTS: Aortic banding was related to an increase in relative weight of the left ventricle from 1.73 +/- 0.06 (sham-operated) to 2.81 +/- 0.25 g/kg, an increase in beta-myosin: alpha-myosin messenger RNA (mRNA) ratio from 0.30 +/- 0.02 to 1.94 +/- 0.55 and an 18-fold increase in left ventricular atrial natriuretic peptide mRNA levels. In contrast, left ventricular pressure overload hypertrophy was not related to a significant change in the abundance of AT1a and AT1b mRNA, which were expressed in a relative ratio of 5:1. Similarly, the abundance of AT2 mRNA was not significantly changed in hypertrophied ventricles. In rats receiving the angiotensin II AT, receptor antagonist losartan (40 mg/kg) for 6 weeks after banding, relative heart weights were 2.39 +/- 0.14 g/kg, the beta-myosin: alpha-myosin ratio was 1.04 +/- 0.20 and atrial natriuretic peptide mRNA levels displayed a blunted increase (11-fold over sham-treated controls), documenting a significant amelioration of left ventricular hypertrophy by blockade of the AT1 receptor. CONCLUSION: Losartan treatment in parallel did not affect AT1a, AT1b and AT2 receptor mRNA levels, which were not different from those in vehicle-treated or sham-treated controls. These findings confirm that left ventricular hypertrophy in the rat is associated with increased ventricular expression of beta-myosin and of atrial natriuretic peptide and with reduced expression of alpha-myosin. Despite these significant changes in cardiac gene expression no alteration was observed in AT1a, AT1b and AT2 receptor mRNA levels. PMID- 8723990 TI - The effect of vitamin B6 on the systolic blood pressure of rats in various animal models of hypertension. AB - OBJECTIVE: To investigate whether a dietary supplement of vitamin B6 could attenuate the elevation of systolic blood pressure (SBP) in Zucker obese or spontaneously hypertensive rats, or rats ingesting sucrose. METHODS: Zucker obese rats (fa/fa), Sprague-Dawley rats with sucrose-induced elevation of SBP, spontaneously hypertensive rats (SHRs) and their corresponding controls were tested for the effects of vitamin B ingestion in different ways: (1) vitamin B6 was included as a supplement (five times the normal intake) from the start of the experiment until the development of hypertension; (2) vitamin B6 supplement was removed from the diet of Zucker obese and Zucker lean control groups after 16 weeks on the dietary treatments; and (3) a diet deficient in vitamin B6 was instituted in SHRs and control Wistar-Kyoto (WKY) rats. The SBP of rats in all groups was monitored in the conscious animal by tail-cuff plethysmography. The effects of the various treatments on the uptake of calcium by caudal artery segments were examined. RESULTS: Male Zucker obese rats (fa/fa) of age 6 weeks fed a commercial rat chow developed hypertension in 3-4 weeks, whereas their lean controls (Fa/Fa) did not. The inclusion of a vitamin B6 supplement (five times the normal intake) resulted in a complete attenuation of the hypertension in the obese strain. Removal of the vitamin B6 supplement from the diet of these obese rats resulted in the return of hypertension within 2 weeks. Similar changes in SBP were also observed in the Zucker lean controls treated with vitamin B6. The ingestion of sucrose by male Sprague-Dawley rats resulted in modest elevation of SBP that was attenuated by the inclusion of the vitamin B6 supplement in their diet. In contrast, there was no response to the inclusion or removal of dietary vitamin B6 supplement in the SHRs. However, the WKY control rats responded to both these conditions in a similar manner to that seen in the Sprague-Dawley strain. Increased peripheral resistance resulting from increased permeability of vascular smooth muscle plasma membrane to Ca2+ is thought to be one of the mechanisms of hypertension. Changes in SBP correlated with changes in the uptake of calcium by caudal artery segments in all the groups studied. The Zucker obese and sucrose-induced hypertensive rats have abnormalities in carbohydrate metabolism. The vitamin B6 supplement decreased the random or fasting blood glucose levels in the Zucker obese and sucrose-fed rats respectively. CONCLUSION: This is the first observation that animal models of hypertension can be classified on the basis of their response to a vitamin B6 supplement. On this basis, the etiology of hypertension in SHRs is quite distinct from that in Zucker obese rats and in rats ingesting sucrose. PMID- 8723991 TI - Mesenteric arterial function in vitro in three models of experimental hypertension. AB - OBJECTIVE: To investigate arterial function in three models of experimental hypertension. METHODS: Twenty-five-week-old spontaneously hypertensive rats (SHRs), deoxycorticosterone-sodium chloride-treated Wistar-Kyoto rats (DOCs), and obese Zucker rats (OZRs) were used. Untreated Wistar-Kyoto rats (WKYs) and lean Zucker rats (LZRs) served as normotensive control rats. The function of mesenteric arterial rings was investigated in organ baths. RESULTS: In norepinephrine-precontracted rings, the relaxations to acetylcholine (ACh) and nitroprusside were reduced in SHRs, DOCs, and OZRs. NG-nitro-L-arginine methyl ester attenuated the dilations to ACh effectively in all study groups, the inhibitory effect being more potent in SHRs and DOCs and similar in OZRs when compared with corresponding controls. Unlike in norepinephrine-precontracted rings, the relaxations to ACh during precontraction with 60 mmol/l potassium chloride (prevention of endothelium-dependent hyperpolarization) were not impaired in the hypertensive animals. Nifedipine inhibited the contractile responses induced by cumulative addition of Ca2+ during stimulation with norepinephrine more effectively in SHRs and DOCs than in WKYs, whereas no such difference was seen between OZRs and LZRs. CONCLUSIONS: Experimental models of genetic, mineralocorticoid-sodium chloride-induced, and obesity-related hypertension were associated with attenuated arterial dilation. The defect of endothelium-mediated relaxation most likely resulted from impaired endothelium dependent hyperpolarization of vascular smooth muscle in these hypertensive models, whereas the endothelial L-arginine-nitric oxide pathway appeared to be preserved. The function of voltage-dependent Ca2+ channels, as evaluated by enhanced inhibitory effect of nifedipine on the arterial contractions, was abnormal in smooth muscle from SHRs and DOCs, whereas such an abnormality was not observed in obesity-related hypertension. PMID- 8723992 TI - Vascular reactivity and flow-pressure curve in isolated kidneys from rats with N nitro-L-arginine methyl ester-induced hypertension. AB - OBJECTIVES: The purpose of this study was to determine the contribution of the functional changes in resistance vessels to the hypertension induced by chronic nitric oxide synthase inhibition in rats. Another goal of this study was to evaluate whether this model of hypertension is accompanied by changes in the activity of endothelium-derived hyperpolarizing factor (EDHF). METHODS: Hypertension was induced by long-term (6 weeks) oral administration of N-nitro-L arginine methyl ester (L-NAME; 75 mg/100 ml in the drinking fluid). Vascular reactivity to vasoconstrictors (phenylephrine and barium chloride) and vasodilators (acetylcholine and nitroprusside) and the flow-pressure curve were examined in isolated perfused kidneys preparations. Vascular reactivity to vasoconstrictors and the flow-pressure curve were studied under basal conditions or after the infusion of L-arginine (100 mumol/l). The activity of EDHF was evaluated by comparing the dose-response curves for acetylcholine obtained in potassium chloride- and phenylephrine-preconstricted preparations. RESULTS: Kidneys from L-NAME-induced hypertensive rats showed increased sensitivity to vasoconstrictors with a greater duration of the pressor responses at high doses and markedly up-shifted flow-pressure curve in comparison with that obtained in control kidneys. These differences disappeared when the kidneys from control and L-NAME-treated rats were infused with L-arginine. The kidneys from L-NAME-treated rats also showed a decreased responsiveness to acetylcholine with an augmented reactivity to nitroprusside. The acetylcholine dose-response curve was reduced in control preparations and greatly attenuated in L-NAME-treated preparations when the renal vasculature was preconstricted with potassium chloride. CONCLUSIONS: The changes in vascular reactivity observed in L-NAME-induced hypertensive rats may play an important role in the pathogenesis of this type of hypertension. Moreover, it is also suggested that long-term nitric oxide inhibition may be associated with increased activity of EDHF. PMID- 8723993 TI - Demonstration of an in vitro direct vascular relaxant effect of diuretics in the presence of plasma. AB - OBJECTIVE: To determine whether diuretics have direct vascular actions and to compare the in vitro effects of a loop diuretic with thiazide and thiazide-like diuretics. DESIGN: A randomized, double-blind, vehicle-controlled design was used to avoid experimenter bias. METHODS: Concentration-response curves to hydrochlorothiazide, chlorthalidone, indapamide, and furosemide were tested on the following male Wistar rat vascular smooth muscle preparations: rat aortic rings, rat pulmonary artery rings, and rat mesenteric portal vein. RESULTS: All four diuretics demonstrated no vasorelaxant action in Krebs solution. They all exhibited vasorelaxant actions in aortic and pulmonary artery rings when plasma was mixed with Krebs solution in a 50:50 ratio. The magnitude of the relaxation was greater in the aortic ring preparation. This direct vascular action was found to be concentration dependent and endothelium independent. The order of potency of the vasorelaxant action of the diuretics was (from most to least) indapamide, hydrochlorothiazide, chlorthalidone, and furosemide on both aortic and pulmonary artery rings. CONCLUSION: Diuretics possess direct vasorelaxant effects that are dependent upon the presence of plasma; this action may contribute to their antihypertensive properties. PMID- 8723995 TI - Relationship of plasma insulin concentration and insulin sensitivity to blood pressure. Is it modified by obesity? AB - OBJECTIVE: To investigate the relationship between insulin level, insulin sensitivity and blood pressure in normoglycaemic men (n = 51) and women (n = 64) aged 53-61 years who were not receiving blood pressure medication and were participants in a previous population-based study. METHODS: Insulin sensitivity was estimated by the minimal model from a frequently sampled intravenous glucose tolerance test. RESULTS: Systolic blood pressure (SBP) did not correlated significantly with fasting insulin level, 2 h insulin level or insulin sensitivity. Diastolic blood pressure (DBP) correlated positively with fasting insulin level but not with 2 h insulin level or insulin sensitivity. However, the positive association between fasting insulin level and DBP was not significant after adjustment for obesity and age. The relationship between high fasting insulin concentration and high DBP was stronger in lean than in obese subjects. The positive correlation between fasting insulin level and DBP was significant in lean but not in obese subjects. CONCLUSIONS: The relationships between decreased insulin sensitivity and compensatory hyperinsulinaemia and blood pressure were rather weak. It is possible that different mechanisms may control blood pressure in lean and obese subjects, with a weaker association between insulin level and blood pressure in obese subjects. Alternatively, in obese subjects long-standing hyperinsulinaemia might increase blood pressure by mechanisms such as sympathetic activation and effects of vasculature, which may mask the underlying contribution of hyperinsulinaemia. PMID- 8723994 TI - Endothelium-dependent relaxation in resistance arteries from spontaneously hypertensive rats: effect of long-term treatment with perindopril, quinapril, hydralazine or amlodipine. AB - OBJECTIVE: To examine the effects of different types of antihypertensive treatment on endothelium-dependent relaxation in resistance arteries from spontaneously hypertensive rats (SHRs). DESIGN AND METHODS: Three-week-old SHRs were treated with the angiotensin converting enzyme (ACE) inhibitor perindopril (1 mg/kg/day) or quinapril (3 mg/kg/day) or the vasodilator hydralazine (50 mg/kg/day) or the calcium antagonist amlodipine (10 mg/kg/day). Control SHRs and Wistar-Kyoto (WKY) rats were treated with water. After 21 weeks rats were culled and mesenteric resistance arteries were mounted in a myograph. Relaxation responses to the endothelium-dependent vasodilators acetylcholine (ACh) and bradykinin were recorded before and after incubation with the nitric oxide synthase inhibitor NG-nitro-L-arginine (L-NOARG), as was the relaxation response to the nitric oxide donor sodium nitroprusside (SNP). RESULTS: All drugs prevented the rise in blood pressure found in the untreated SHRs. ACh-induced relaxation was significantly impaired in the untreated SHRs compared with the WKY rats. Treatment with either ACE inhibitor prevented the development of this impaired response. ACE inhibitor treatment significantly increased the relaxation response to bradykinin. Despite lowering blood pressure, hydralazine or amlodipine had no effect on ACh- or bradykinin-induced relaxation. Responses to SNP were not different between untreated SHRs and WKY rats and were not affected by drug treatment. CONCLUSION: Specific properties of certain antihypertensive drugs may play an important role in restoring endothelium-dependent relaxation in the small arteries that regulate peripheral resistance in the SHR. PMID- 8723996 TI - Oxidative stress and hyperinsulinaemia in essential hypertension: different facets of increased risk. AB - OBJECTIVE: To evaluate oxidative stress markers in normo- and hyperinsulinaemic essential hypertension patients, and to relate these parameters to plasma glucose and insulin levels. METHODS: Diene conjugates, thiobarbituric acid reactive substances, iron-stimulated thiobarbituric acid reactive substances and anti oxidative capacity of serum were detected in 32 untreated essential hypertension patients with normal glucose tolerance, divided into hyperinsulinaemic (n = 12, fasting plasma insulin level > 13.5 mU/l, means 2 SD of controls) and normo insulinaemic (n = 20) subgroups, compared with 26 age- and body mass index matched controls. Plasma glucose and insulin levels were measured during an oral glucose tolerance test. RESULTS: Levels of lipid peroxidation products (diene conjugates, thiobarbituric acid reactive substances and iron-stimulated thiobarbituric acid reactive substances) were elevated and serum anti-oxidative capacity decreased both in hyper- and in normo-insulinaemic patients compared with those in controls, with no significant differences between the hypertensive subgroups. No independent correlations were detected between oxidative stress markers and fasting or stimulated plasma insulin and glucose levels. The essential hypertension patients were characterized by a lower fasting glucose: insulin ratio and enhanced plasma insulin response to oral glucose test compared with controls. CONCLUSIONS: The results suggest that oxidative stress occurs, in addition to disturbances in glucose metabolism, in essential hypertension patients, thus potentially exposing them to increased risk of developing complications. Factors other than plasma insulin level are likely to contribute to oxidative stress in hypertensive patients with normal glucose tolerance. PMID- 8723997 TI - Methodological aspects in the assessment of circadian blood pressure variation in the elderly. PMID- 8723998 TI - The use of embryo genotyping in the propagation of genes involved in the immune response. AB - Multiple ovulation and embryo transfer (MOET) now enables researchers to produce identical twin animals, to obtain progeny from pre-pubertal females and to obtain more offspring from valuable animals. MOET and sexed semen have produced genetic progress of up to 60% of milk production. The oestrous cycles of animals are synchronized with progestagens before superovulation with gonadal hormones, pregnant mare serum gonadotrophin and follicle stimulating hormone. Surgical, non surgical and laparoscopic methods are applied to recover and transfer embryos. Sexing and genotyping of the pre-implantation embryos is a key step in improving the management and breeding programmes for livestock, as well as in the human for the prenatal diagnosis of genetic disorders. Several serological and physiological methods have been used to determine the sex of the pre-implantation embryos; none has had satisfactory results in terms of time and accuracy. Sexing by polymerase chain reaction (PCR) using male-specific chromosome sequences alone or with female-specific chromosomal DNA probes simultaneously has been sufficient to identify the sex of the embryos with 100% accuracy. However, caution should be taken against sources of the contamination. The MHC class I, class II and background genes have been implicated in resistance to internal parasites in animals. Biotechnological methods such as screening of embryos prior to transfer using PCR and primer extension pre-amplification have already made it possible to detect transgenic or genetically disordered embryos and could be applied to select those embryos bearing immunological genotypes of interest, such as resistance to internal parasites. Ultimately, cloning and nuclear transplantation would provide the possibility of isolating these resistance genes and to transfer them to livestock pre-implantation embryos to propagate these desirable traits. PMID- 8723999 TI - Transcription factor AP-1, and the role of Fra-2. AB - Transcription factors function to regulate gene transcription. They may be constitutively expressed or may only be activated during specific situations. Activator protein-1 (AP-1) is an inducible transcription factor, and is comprised of multiple protein complexes that include the gene products of the fos and jun gene families. Numerous cellular and viral genes contain AP-1 binding sites within their promoters and, accordingly, AP-1 has been shown to play a role in the regulation of both basal and inducible transcription of these genes. fos related antigen-2 (fra-2) has been found to have both similar and unique properties to that of other fos gene members in terms of its regulation and expression. The analysis and determination of the function of Fra-2 will provide further information on the role of AP-1. PMID- 8724000 TI - Gonadotropin-releasing hormone agonist influences absolute levels of lymphocyte subsets in vivo in male mice. AB - Our earlier studies have demonstrated that gonadotropin-releasing hormone (GnRH) agonists suppress immune system function in female mice. No systematic studies regarding the effect of gender on immune system function following GnRH agonist treatment, however, have been reported. This study, therefore, investigated sequential changes in lymphocyte subsets in 3- and 10-week-old male mice following agonist or placebo administration. Changes in immunophenotypic expression of lymphocytes from thymus, bone marrow, spleen, and blood were analysed at periodic intervals. Upon agonist administration, plasma testosterone levels were significantly increased in pre-pubertal mice, but were significantly decreased in post-pubertal males. Absolute thymic weights, thymocytes and T subsets were significantly increased from the third week regardless of gonadal status. Blood lymphocyte subsets showed a decreasing trend after agonist administration in pre-pubertal males, whereas no differences were observed in post-pubertal males. No significant differences were observed in spleen cells after agonist administration. These studies, together with earlier observations in female mice indicate that GnRH agonist effects on the immune system, are independent of steroid hormone levels. In contrast to suppressive effects in females, GnRH agonist induce no change or ultimately enhanced lymphocyte counts in males, indicating differential effects on the immune system between males and females. This may have important implications for the treatment of various diseases. PMID- 8724001 TI - Control of scarring in adult wounds using antisense transforming growth factor beta 1 oligodeoxynucleotides. AB - Adult wounds heal with scar formation, whereas fetal wounds heal without scarring and with a lesser inflammatory and cytokine response. We reasoned that a strategy employing antisense oligodeoxynucleotides (ODN) complementary to transforming growth factor (TGF)-beta-1 mRNA might decrease the scarring of dermal wounds in the mouse. To evaluate this concept, we tested the effects of antisense ODN targeted to TGF-beta 1 mRNA by topical application of the ODN on the skin wound. Phosphorothioation of ODN to retard their degradation. When antisense TGF-beta 1 ODN were applied on the wound site, there was marked reduction of scarring compared with a control wound site. This effect of antisense TGF-beta 1 ODN on scar formation was associated with decreased expression of the TGF-B1 gene. However, sense TGF-beta 1 ODN had no effect on the expression of the TGF-beta 1 gene. In addition, control wounds healed with excessive fibrosis compared with the antisense-treated wounds. In conclusion, our results indicate that antisense TGF-beta 1 ODN could be used for ameliorating scar formation during wound healing. PMID- 8724003 TI - Uncontrolled triggering of programmed cell death (apoptosis) in haematopoietic stem cells: a new hypothesis for the pathogenesis of aplastic anaemia. AB - The cause of bone marrow failure in aplastic anaemia (AA) is still unknown; however, it is clear that acquired AA is a heterogeneous disease including basically different pathophysiological conditions. Causative agents, clinically associated with AA, possibly exert their action through restricted pathways. Some theoretical and experimental data show that programmed cell death (PCD) or apoptosis is physiologically important in normal haematopoiesis and could be involved in the pathophysiological events responsible for the development of AA. Therefore, it is intriguing to hypothesize that the pathogenetic mechanism underlying most cases of acquired AA could be represented by an excessive and/or uncontrolled triggering of PCD in haematopoietic stem cells. Investigations to test this hypothesis are proposed. PMID- 8724002 TI - Molecular cloning and characterization of tumor necrosis factor alpha (TNF-alpha) from the Australian common brushtail possum, Trichosurus vulpecula. AB - Immune responses in the Australian common brushtail possum (Trichosurus vulpecula) and in particular the role of cytokines are poorly understood. We have undertaken to isolate cytokine genes using reverse transcriptase-polymerase chain reaction (RT-PCR) and in this study describe the molecular cloning of TNF-alpha. Primers were designed from consensus sequences at the N-terminus end of eutherian mammalian TNF-alpha and the possum cDNA, derived from spleen RNA, identified by RT-PCR. The complete cDNA encoding possum TNF-alpha was amplified from lymphocyte RNA by 5' and 3' rapid amplification of cDNA ends (RACE). The nucleotide sequence of the protein coding region of this cDNA shared 66-69% identity with other mammalian TNF-alpha genes. The predicted protein of 233 amino acids shared 56-58% identity with eutherian mammalian TNF-alpha was expressed in both Saccharomyces cerevisiae and Escherichia coli by constructing expression plasmid derivatives of the vectors pYES2 and pGEX-2T respectively. Cell extracts prepared from transformants and the purified GST/TNF-alpha fusion protein exhibited cytotoxic activity on the TNF-alpha-sensitive murine fibroblast L929 cells and stimulated proliferation of possum thymocyte cells. The induction of possum TNF-alpha mRNA in alveolar macrophages was analysed by RT-PCR using possum-specific TNF-alpha primers. Macrophages cultured in the presence of LPS showed enhanced transcription of TNF-alpha mRNA. This is the first report of the cloning and sequence analysis of the cDNA encoding a marsupial cytokine gene. PMID- 8724005 TI - Antigen presentation in allergic sensitization. AB - IgE antibodies, when cross-linked by allergen on the surface of effector cells such as mast cells and basophils, are known to be directly responsible for immediate type hypersensitivity reactions. In addition, IgE may be involved in other, indirect, mechanisms, fundamental to the pathogenesis of allergic diseases, such as enhancement of the antigen capturing capacity of antigen presenting cells. IgE mediated antigen presentation could lead to a continuous activation of the immune system by very low concentrations of allergen. As a result, Th2 cell populations may expand and may induce more B cells to switch to IgE production. Subsequently, the overproduction of IgE and Th2 cells in a patient may explain the clinical observation that certain allergic patients deteriorate from sensitivity to a single group of allergens to sensitivity to multiple groups of allergens. Therefore, control of IgE production is not only important for the treatment of allergic symptoms, but may also regulate deterioration of allergy via the mechanism of CD23/IgE mediated allergen presentation by naive B cells. The role that monocytes, which have recently been found to express Fc epsilon RI, play in the pathogenesis of allergy, remains speculative. We hypothesize that their role may be to remove IgE from the circulation and re-direct the immune response from naive B cells. IgG antibodies which cannot be used for antigen uptake by B cells also direct the immune response to monocytes. PMID- 8724004 TI - Populations of thymocytes and peripheral blood leucocytes in leukaemia-bearing mice treated with G-CSF. AB - The influence of granulocyle-colony stimulating factor (G-CSF) on thymocyte subsets and peripheral blood leucocytes in leukaemia L1210-bearing mice was evaluated. Leukaemia-bearing mice have a markedly reduced L3T4+Lyt2+ thymocyte subpopulation. We observed "correction" of relative values of thymocyte subpopulations induced by treatment of mice with a G-CSF preparation. The suggestion is presented that this could result from liberation of lymphocyte T precursor cells from the bone marrow and increased homing of them into the thymus. The direct or indirect influence of G-CSF on the L3T4+Lyt2+ subpopulation of thymocytes is discussed. PMID- 8724006 TI - Immunologic responses following respiratory sensitization to house dust mite allergens in mice. AB - Allergens from the house dust mite Dermatophagoides pteronyssinus are a major cause of human respiratory diseases, including asthma. In order to help in understanding the early events in allergen sensitization, a murine model of allergic respiratory disease was developed. Mice were immunized by intranasal inoculation of Der p 1 or Der p 2 on days 0, 3, 7, 10, 14, 17, 21 and 29. T cell reactivity was determined using in vitro assays of allergen-specific cytokine production by cells from the spleen, the draining superficial cervical lymph nodes (SCLN) and the non-draining brachial and inguinal nodes. The cytokines assayed in supernatants were IL-4, as a measure of Th2 activation, IL-2 as a measure of Th 1 activation, and IL-3/GM-CSF as an overall marker of T cell stimulation. There was evidence of local and systemic T cell activation by day 7, with the release of IL-2 and of IL-3/GM-CSF by SCLN and spleen cells, respectively. IL-4 production by SCLN and spleen cells was not evident until day 21. T cell sensitization in non-draining node groups was not detected. Intradermal skin tests were performed at the above specified times and positive wheal responses indicated that specific IgE was present from day 3. The above data suggest that respiratory immunization to allergen is rapid and is associated with early systemic sensitization. In this model both Th1 and Th2 responses were evident, with the Th1 response occurring early and the Th2 following after repeated immunizations. PMID- 8724007 TI - Peptide-mediated regulation of the allergic immune response. AB - A major key to successful immunotherapy may depend on altering the qualitative nature of the immune response in allergic patients. In this review we examine how immune responses to environmental allergens are regulated, and the mechanisms used by the immune system to prevent allergic sensitization. We also discuss future prospects of using allergen-derived peptides in immunotherapy and the possibility of 'reprogramming' the immune responses by immunizing under conditions that promote Th1 responses instead of Th2 responses. PMID- 8724008 TI - Immunological and structural similarities among allergens: prerequisite for a specific and component-based therapy of allergy. AB - It is known that allergic patients are frequently co-sensitized against different allergen sources. Progress made in the field of allergen characterization by molecular biological techniques has now revealed that sensitization against different allergen sources can be explained as cross-reactivity of IgE antibodies with structurally and immunologically related components present in these allergen sources. This review defines groups of cross-reactive plant allergens with significant sequence homology. The similarities among allergens may facilitate allergy diagnosis by using a few representative cross-reactive allergens to determine the patient's IgE reactivity profile (allergogram). According to that typing, a few cross-reactive allergens, carrying most of the relevant IgE epitopes, may then be selected for patient-tailored specific therapy. PMID- 8724009 TI - Anti-IgE in allergic sensitization. AB - Anti-IgE autoantibodies exist predominantly in the sera of patients with atopic disease. For some time such anti-IgE autoantibodies have been considered a phenomenon that may not be of clinical importance. The cloning of such anti-IgE autoantibodies has eliminated doubts of whether these antibodies exist, but it is still unclear whether such autoantibodies play a pathophysiological role. However, there are ongoing clinical trials that use humanized anti-IgE antibodies for passive immunization of atopic individuals. While this approach may not definitely clarify the role of anti-IgE autoantibodies, it will nevertheless clarify the role of IgE. PMID- 8724010 TI - Atopy and the ectopic immune response. AB - A hypothesis if formulated for the mode of action of the 'atopy gene': the gene(s) causing an increased tendency to mount an IgE immune response to trivial environmental antigens. It is postulated that some antigens are not efficiently transported to the lymphoid organs, the location of the 'eutopic' immune system; migratory lymphoid cells may occasionally react and start an inefficient immune response in sites not so destined. This is called an ectopic immune response. This response lacks the regulatory influences prevailing in the lymphoid organs, which results in approximately equal quantities of the various immunoglobulin isotypes, including IgE. The atopy gene(s) work by increasing the risk of such an ectopic immune response. PMID- 8724011 TI - The role of Th2 type CD4+ T cells and Th2 type CD8+ T cells in asthma. AB - The cellular inflammatory responses which are seen in allergic and asthmatic diseases are viewed as being quite strongly dependent on the activities of T cells and their products. The major T cell subset involved appears to be the so called CD4+ Th2 subset which produces interleukin-4 (IL-4) and interleukin-5 (IL 5). In vitro and in vivo experiments have indicated that IL-4 is a key regulator in these kinds of immune responses, not only switching B cells to IgE production, but acting on CD4+ T cells to drive their development towards a Th2 phenotype. Recent results have shown that the functional phenotype of CD8+ T cells can be switched from interferon gamma production to IL-4 and IL-5 production by the presence of IL-4. This could prove an especially important phenomenon since it is the production of interferon gamma by CD8 T cells which is seen as necessary for protection against virus infection. This short review updates our current knowledge of how IL-4 can act on CD4+ and CD8+ T cell subsets in in vivo models of asthma and allergic disease. PMID- 8724013 TI - How is expression of the interleukin-5 gene regulated? AB - Eosinophilia is a uniquely specific phenomenon regulated by interleukin-5 (IL-5), suggesting specific control for IL-5 expression. However in eosinophilia IL-5 is often co-expressed with other lymphokines such as IL-4, indicating that common, as well as independent, control mechanisms must exist. IL-5 gene expression is regulated at the transcriptional level. The molecular analysis of the IL-5 promoter region reveals the presence of positive regulatory sites that are common to many lymphokine genes. Results from immunosuppression studies suggest that the key control mechanism of IL-5 regulation may not depend on specific regulatory factors but on how gene expression is activated. PMID- 8724012 TI - The relevance of murine animal models to study the development of allergic bronchial asthma. AB - Bronchial asthma (BA) develops on the basis of a genetic predisposition and involves a characteristic sequence of changes in immune functions. In the immunopathogenesis, several phases can be distinguished: the initial stage is defined as the development of allergic sensitization. This step is dependent on: (i) T cell activation; (ii) IL-4 production; (ii) IgE synthesis; and (iv) mediator release by effector cells. The second phase of allergic inflammation as a consequence of the T cell dependent sensitization is characterized by IL-5 production and eosinophil activation and recruitment. Airway mucosa remodelling is the consequence of chronic inflammatory processes and represents the final stage of BA. In this article animal models will be discussed with regard to their relevance for these different phases in development of chronic allergic BA. PMID- 8724014 TI - The structure and function of hyaluronan: An overview. AB - Hyaluronan is a major component of synovial tissue and fluid as well as other soft connective tissues. It is a high-Mr polysaccharide which forms entangled networks already at dilute concentrations (< 1 mg/mL) and endows its solutions with unique rheological properties. Physiological functions of hyaluronan (lubrication, water homeostasis, macromolecular filtering, exclusion, etc.) have been ascribed to the properties of these networks. Recently a number of specific interactions between hyaluronan and a group of proteins named hyaladherins have also pointed towards a role of hyaluronan in recognition and the regulation of cellular activities. Many more or less well documented hypotheses have been proposed for the function of hyaluronan in joints, for example, that it should lubricate, protect cartilage surfaces, scavenge free radicals and debris, keep the joint cavities open, form flow barriers in the synovium and prevent capillary growth. PMID- 8724015 TI - HMG-CoA reductase inhibitors and bile lithogenicity: cholesterol supersaturation or nucleation defect? PMID- 8724016 TI - Does the HMG-CoA reductase inhibitor pravastatin influence nucleation of cholesterol crystals in supersaturated model bile? AB - OBJECTIVE: To assess whether the presence in bile of HMG-CoA reductase inhibitors, which are secreted predominantly into the bile, influences biliary lithogenicity. DESIGN: Physiologic biliary concentrations of the hydrophilic HMG CoA reductase inhibitor pravastatin were added to supersaturated model bile (cholesterol saturation index 1.4) and vesicles, the latter with and without the concomitant addition of the nucleation-promoting bile salt taurodeoxycholate. OUTCOME MEASURES: Nucleation time, defined as the number of days after which cholesterol monohydrate crystals are visible by phase contrast microscopy in filtered samples of model bile and vesicles, was assessed. RESULTS: The addition of pravastatin 0.01-1 mg/ml did not influence the nucleation time of supersaturated model bile (mean nucleation time without pravastatin: 8.3 +/- 2.2 days (SD), with pravastatin 1 mg/ml 9.3 +/- 0.4 days and pravastatin 0.01 mg/ml 7.6 +/- 2.3 days). The addition of similar concentrations of pravastatin to vesicle fractions alone did not influence nucleation time (> 20 days), nor could it prevent the nucleation-promoting effect of taurodeoxycholate (nucleation time with or without pravastatin 1 day). CONCLUSION: The results from this in-vitro study indicate that the presence of pravastatin in bile may not influence gallbladder bile lithogenicity. It can be hypothesized that this also applies to other HMG-CoA reductase inhibitors. PMID- 8724017 TI - 24-hour oesophageal motility in gastro-oesophageal reflux disease (GORD): increased occurrence of simultaneous contractions. AB - Impaired oesophageal peristalsis may play a major pathogenetic role in gastro oesophageal reflux disease (GORD). Therefore 55 patients with suspected GORD were studied simultaneously by ambulatory 24-hour pH and pressure monitoring with three pressure transducers (3, 8 and 13 cm above the lower oesophageal sphincter) in order to to test for a relationship between oesophageal motility and GOR. Twenty-one patients (38%) had pathological reflux; these patients had significantly more simultaneous contractions than patients without pathological GOR (30.1 +/- 3.3% vs. 19.0 +/- 1.8%, P = 0.002, mean +/- SEM). Further analysis revealed a significant difference between groups in the occurrence of simultaneous contractions in the mid-oesophagus (33.7 +/- 3.8% vs. 23.9 +/- 1.8%, P = 0.012), but not in the distal oesophagus (34.4 +/- 2.7% vs. 33.9 +/- 3.1%, P = 0.90). In addition, a moderate but highly significant correlation between the rate of simultaneous contractions and reflux time was found (r = 0.463, P = 0.0005). Mean amplitude and mean duration of the contractions were no different between groups, neither in the proximal (43.4 +/- 3.3 mmHg vs. 44.9 +/- 1.9 mmHg, P = 0.68 and 2.4 +/- 0.2s vs. 2.5 +/- 0.1s, P = 0.50, respectively) nor in the distal oesophagus (48.8 +/- 4.6 mmHg vs. 54.2 +/- 3.4 mmHg, P = 0.34 and 3.0 +/- 0.2s vs. 2.9 +/- 0.2s, P = 0.71, respectively). It was concluded that pathological GOR is associated with an increased occurrence of simultaneous contractions in the mid, but not in the distal, oesophagus. PMID- 8724018 TI - Oesophageal acid clearance in patients with systemic sclerosis: effect of body position. AB - OBJECTIVE: To test the effect of body position on oesophageal acid clearance time in patients with systemic sclerosis. DESIGN AND METHODS: Fifteen consecutive patients with systemic sclerosis and six healthy subjects underwent oesophageal manometry and an acid clearance test in three body positions (supine, recumbent at 30 degrees and seated at 90 degrees) in randomized order. RESULTS: In healthy subjects the body position did not affect acid clearance time, whereas in patients the oesophagus cleared mainly by gravity. In patients the acid clearance time was significantly longer in the supine than in the seated position (P < 0.05). Nine patients did not have a detectable peristaltic wave in the distal oesophagus. In the other six oesophageal peristalsis was still detectable but contractions had reduced amplitude and often had double and triple peaks; also in this subgroup the acid clearance time recorded in the supine position was prolonged. CONCLUSIONS: In systemic sclerosis gravity plays a major role in oesophageal acid clearance time. The finding of delayed acid clearance in a supine patient may suggest initial oesophageal involvement in the disease. PMID- 8724019 TI - Hyperglycaemia affects proximal gastric motor and sensory function in normal subjects. AB - OBJECTIVE: Hyperglycaemia delays gastric emptying in normal subjects and patients with diabetes mellitus by uncertain mechanisms and may affect the perception of somatic sensations. The effects of hyperglycaemia on the motor function of the proximal stomach and the perception of gastric distension were evaluated in normal subjects. DESIGN: Paired studies were performed in randomized order in 10 healthy volunteers on separate days during euglycaemia and hyperglycaemia (blood glucose approximately equal to 15 mmol/l). METHODS: With a barostat and a balloon positioned in the proximal stomach, tasting subjects underwent a stepwise gastric distension. Each 2 mmHg step was maintained at a constant pressure for 2 min. The volume of the barostat balloon was measured and perception of the sensations of fullness, desire to belch, nausea, abdominal discomfort and hunger was scored at each step. RESULTS: Hyperglycaemia was associated with an increase in proximal gastric compliance (P < 0.01) evident from 2 mmHg above basal intragastric pressure. Perception scores for the sensations of nausea and desire to belch were greater during hyperglycaemia than euglycaemia (P < 0.05) in relation to both pressure at each step and volume. Hyperglycaemia did not affect perception of the sensations of abdominal discomfort, fullness or hunger. CONCLUSIONS: Hyperglycaemia increases proximal gastric compliance, reflecting a reduction in gastric tone. This may contribute to the previously observed delay in gastric emptying associated with hyperglycaemia. Hyperglycaemia appears to increase the perception of some of the sensations induced by gastric distension. PMID- 8724020 TI - Follow-up of treated coeliac patients: sugar absorption test and intestinal biopsies compared. AB - OBJECTIVE: To determine whether the sugar absorption test (SAT) during follow-up of patients with coeliac disease on a gluten-free diet (GFD) correlates with improvement of the villous architecture of the small intestine. METHODS: The SAT was performed in coeliacs at diagnosis and during follow-up with GFD. For the SAT, a solution of lactulose (L) and mannitol (M) was given to the fasting patient and the L-M ratio calculated in a 5-hour urine sample by gas chromatography: ratios > 0.089 are considered abnormal. The solution was made hyperosmolar by adding sucrose (1560 mmol/l). RESULTS: The L-M ratio was 2-3 times higher at diagnosis than either at 8 months to 2 years gluten free, or beyond 2 years gluten free, consecutively. The L-M ratio (mean, range) was significantly higher in cases of biopsies with (sub)total villous atrophy (VA) (0.388, 0.062-0.804, n = 28), partial VA (0.240, 0.062-0.841, n = 18) and villous irregularity (0.143, 0.017-0.322, n = 29) than in case of normalized histology after GFD (0.085, 0.021-0.230, n = 19). The rate of normalization of functional integrity was slower in adults than in children, demonstrated by a combination of histology and SAT. CONCLUSION: The SAT correlates well with the degree of VA. It is important for daily clinical practice that the simple and non-invasive SAT can be used as an indicator of intestinal damage, thus influencing need for and timing of intestinal biopsies. PMID- 8724021 TI - Effect of cigarette smoking on the course of Crohn's disease. AB - OBJECTIVE: It has been shown that smokers are more likely than non-smokers to develop Crohn's disease. In order to examine the influence of smoking on the course of Crohn's disease, as measured by the risk of surgery, the need for surgery was assessed in patients with Crohn's disease. DESIGN: The course of Crohn's disease was evaluated as to smoking status by life table analysis. SETTING: A tertiary hospital in Essen, Germany. PATIENTS: The study involved 346 patients with Crohn's disease admitted to hospital between 1989 and 1992. Only patients residing in the City of Essen were included. RESULTS: Of the 346 patients, 144 smokers and 143 non-smokers were included in the analysis. Overall, 73% of smokers and 39% of non-smokers required one or more operations. The corresponding relative risks (RR) and 95% confidence intervals were for one operation and more than one operation or no operation at all (RR 1.0) 3.9 (2.2 6.9) and 10.8 (5.3-22.1), respectively. There were significant differences as to recurrence rates (defined as further surgery after first surgery for control of disease) between smokers and non-smokers. For smokers the 5- and 10-year recurrence rates were 43% and 64%. For non-smokers corresponding recurrence rates were 26% and 33%. For 5- and 10-year recurrence rates the RR estimates for smokers versus non-smokers were 3.1 (1.7-5.8) and 6.7 (2.7-6.8). When stratified by gender, the increased risk for recurrence was obvious in both sexes. For the number of cigarettes smoked a dose-response effect was obvious in women. CONCLUSION: These data suggest that the course of Crohn's disease is less favourable in smokers than in non-smokers. PMID- 8724022 TI - Placebo-controlled clinical trial of mesalazine in the prevention of early endoscopic recurrences after resection for Crohn's disease. Groupe d'Etudes Therapeutiques des Affections Inflammatoires Digestives (GETAID). AB - OBJECTIVE: Endoscopic postoperative recurrences occur early after 'curative' surgery for Crohn's disease. Pentasa has been shown to be effective in the maintenance treatment of quiescent Crohn's disease. The aim of this study was to test the efficacy of a 12-week oral intake of Claversal in the prevention of endoscopic recurrences after 'curative' resection for ileal, colonic or ileocolonic Crohn's disease. We conducted a multicentre double-blind controlled trial comparing Claversal (1g tid) with placebo, starting within 15 days after surgery. The macroscopic normality of the two anastomotic segments was assessed at surgery. Patients were clinically and biologically evaluated twice (6-week interval), and colonoscopy was performed at 12 weeks. Endoscopic relapse was defined by any anastomotic ulcerations or stenosis and staged according to a four grade score. RESULTS: Between May 1989 and May 1991 12 centres included 126 patients, 70 women and 56 men, aged 33 +/- 12 years (range 16-70) in the study. Disease locations were ileal, colonic and ileocolonic in 45, 6 and 49%, respectively. Claversal and placebo groups were similar at inclusion, except for ESR (37 +/- 26 vs. 27 +/- 23 mm/h in the Claversal and placebo groups, respectively; P < 0.05). Nine patients were withdrawn from the study. Adverse reactions occurred only in six patients. Five patients were excluded for protocol violation. Finally, 106 patients could be evaluated at 12 weeks (55 Claversal and 51 placebo). An endoscopic relapse was observed in 50% and 63% of the Claversal and placebo groups, respectively (P = 0.16), with a similar grade distribution. Claversal was well tolerated. CONCLUSIONS: Our study confirms that a large proportion of endoscopic recurrences occur within 3 months of resection in Crohn's disease. There was a slight trend towards greater efficacy of Claversal; it could be worthwhile trying higher dosages and/or 5-ASA compounds with different intestinal release profiles. PMID- 8724023 TI - Faecal incontinence with transitory absence of anal contraction in two sexually or physically abused women. AB - OBJECTIVE: To report two cases of faecal incontinence caused by transitory absence of anal voluntary contraction without anal anomalies. PATIENTS: Two women referred to our gastroenterology department with faecal incontinence. One patient had been sexually abused by her father; the other had been physically abused by her husband. RESULTS: Clinical and manometric anal contraction was absent despite normal anal endosonography and normal electrophysiological perineal study. Anal contraction was completely normalized after biofeedback, and the patients recovered from the faecal incontinence. CONCLUSION: Transitory absence of anal contraction is very uncommon as the origin of faecal incontinence. It was observed in two abused women. PMID- 8724024 TI - Endoscopic and histological appearance of pancreatic metaplasia in the human gastric mucosa: a preliminary report on a recently recognized new type of gastric mucosal metaplasia. AB - Recently a new type of gastric mucosal metaplasia referred to as pancreatic metaplasia (pancreatic acinar metaplasia) has been recognized. The difference is characterized by the presence of epithelial cells with the cytoplasm strongly basophilic in the basal compartment, whereas in the middle and apical portions it is acidophilic and finely granular. In contrast to intramuscular heterotopic pancreas, foci of pancreatic metaplasia are located in the lamina propria of the gastric mucosa. They form lobules, small clusters, or may be single. The present series included 13 cases of pancreatic metaplasia found in gastroscopic specimens obtained from seven women and six men. We investigated the histological pattern and endoscopic changes. Based upon clinical data it was found that in five patients the gastric mucosa was defined as 'dichromatic'. It is possible that studies involving greater numbers of patients will permit closer correlation between the histological pattern of pancreatic metaplasia and the corresponding endoscopic changes. PMID- 8724025 TI - Long-term clinical outcome of post-cholecystectomy patients with biliary-type pain: results of manometry, non-invasive techniques and endoscopic sphincterotomy. AB - OBJECTIVE: The purpose of the present study was to investigate prospectively the value of manometry, non-invasive techniques and endoscopic sphincterotomy in biliary type II and III patients with suspected sphincter of Oddi dysfunction. METHODS: All patients (n = 31) had undergone a cholecystectomy from 1 to 45 years before entry into the study, and had a history of biliary-type pain ranging from 4 to 156 months. In a primary diagnostic work-up, including ERCP, other causes of gastrointestinal disease were excluded. The width of the common bile duct, pain induced by the injection of contrast medium into the common bile duct, delayed contrast drainage and the results of a morphine-neostigmine test were analysed. Manometric measurements of basal sphincter of Oddi pressure were performed. Twenty-three patients with an elevated basal pressure (> 40 mmHg) underwent endoscopic sphincterotomy. RESULTS: During the follow-up period (8-62 months) 83% of the patients became symptom free or experienced an improvement. Basal sphincter of Oddi pressure and long-term clinical outcome of patients correlated to different degrees with the results of non-invasive techniques. CONCLUSIONS: Our data suggest that endoscopic sphincterotomy may be an effective therapeutic modality in group II and III patients with sphincter of Oddi dysfunction in whom other gastrointestinal disorders have previously been excluded. PMID- 8724026 TI - Do patients with sphincter of Oddi dysfunction benefit from endoscopic sphincterotomy? A 5-year prospective trial. AB - OBJECTIVES: To assess the incidence of elevated sphincter of Oddi baseline pressure and the response to endoscopic sphincterotomy in patients with suspected sphincter of Oddi dysfunction. DESIGN: A 5-year prospective clinical trial. METHODS: One-hundred and eight patients with recurrent biliary-type pain after cholecystectomy were enrolled. After thorough investigation, 35 patients with suspected type II sphincter of Oddi dysfunction (SOD) and another 29 type III patients remained for further investigation. Both groups were similar with respect to demographic data and severity of pain. Biliary manometry was performed in all except three patients in either group. Endoscopic sphincterotomy was performed in all patients with abnormal sphincter of Oddi baseline pressure (> 40 mmHg). All patients were clinically re-evaluated after 4-6 weeks, and thereafter the sphincterotomized patients were followed for a median period of 2.5 years. RESULTS: An abnormal sphincter of Oddi baseline pressure was found in 62.5% of the type II patients and in 50% of the patients with suspected type III SOD (P = 0.66). At the 4-6 week follow-up none of those patients without abnormal manometry, but 70% of the patients with type II SOD, and 39% of the type III SOD patients, respectively, reported subjective benefit after sphincterotomy (P = 0.13 type II vs. type III). However, after a median follow-up of 2.5 years, sustained symptomatic improvement after sphincterotomy was found in 60% of the type II patients, but only in 8% of the patients with type III SOD (P < 0.01). CONCLUSION: Disregarding a lack of difference in the incidence of abnormal sphincter of Oddi baseline pressure between type II and type III SOD, the Geenen Hogan classification helps to predict the clinical outcome after endoscopic sphincterotomy. PMID- 8724027 TI - Splenic hypofunction in cirrhosis is not associated with increased risk for infections. AB - BACKGROUND/AIM: Pitted cell count has been described as a sensitive marker of splenic function. Recently, pitted cell count was shown to be increased in patients with alcoholic liver disease, and also to be associated with an increased susceptibility to infection. Therefore, our aim in the present study was to assess splenic function in a group of patients with cirrhosis and to determine its possible role in the development of infections. METHODS: Splenic function was assessed during hospitalization in 44 patients diagnosed as having cirrhosis, and was compared to 18 healthy subjects. Function was evaluated by counting the number of pitted cells in peripheral blood films. Results were the mean value of two independent counts and were expressed as number of pitted cells/100 erythrocytes. RESULTS: The mean percentage of pitted cells in peripheral blood of cirrhotic patients was significantly greater than that assessed in controls (3.5 +/- 3.7% vs. 1.6 +/- 0.9%, P < 0.01, Mann-Whitney U test). Eighteen patients with cirrhosis (36%) had counts greater than 3.4% (mean + 2 SD of the control group). There were no significant differences between patients with counts above or below 3.4% in terms of liver tests, platelet and reticulocyte counts, the presence of complications, Child-Pugh score and the prevalence of infections. There was no difference in pitted cell count between alcoholic and non-alcoholic cirrhotic patients (3.8 +/- 3.4% vs. 3.1 +/- 2.8% respectively, P, NS). Moreover, the mortality rate and the occurrence of infections were similar in patients with normal and increased pitted cell counts during 1-year follow-up. CONCLUSION: The increased number of pitted cells in patients with cirrhosis is not related to heptic function and does not represent a risk factor for infections. PMID- 8724028 TI - Cerebral blood flow, oxygen metabolism and transcranial Doppler sonography during high-volume plasmapheresis in fulminant hepatic failure. AB - OBJECTIVE: The effect of high-volume plasmapheresis on hepatic encephalopathy, cerebral blood flow (CBF) and cerebral metabolic rate for oxygen (CMRO2) was investigated in patients with fulminant hepatic failure (FHF). METHODS: Twelve consecutive patients (8 women, 4 men, median age 34 years (range 19-51), were studied before and after high-volume plasmapheresis with 10-16 litres fresh frozen plasma, while PaCO2 and body temperature were maintained at 30 (23-34) mmHg and 37.6 degrees C (36.6-38.4), respectively. Blood samples from the internal jugular vein and a radial artery allowed calculation of the cerebral arteriovenous oxygen difference (AVDO2) and oxygen extraction (AVDO2 divided by arterial oxygen content). CBF was determined by a xenon-133 clearance method in eight patients and CMRO2 calculated as AVDO2 times CBF. Cerebral perfusion pressure (CPP) was determined as the difference between mean arterial and subdural pressures in eight patients. RESULTS: High-volume plasmapheresis was initiated 22 (6-168) h after the development of hepatic encephalopathy and 11 patients had grade 4 encephalopathy. Following high-volume plasmapheresis the grade of encephalopathy improved in four patients. The CBF increased from a median of 31 (16-86) to 45 (18-97) ml/100 g/min and as oxygen extraction remained unchanged (32 (9-41) vs. 29 (7-39)%), CMRO2 increased from 1.24 (0.96-1.82) to 1.86 (1.00-2.07) ml/100 g/min (P < 0.05). The CPP increased from 62 (19-76) to 92 (50-105) mmHg (P < 0.01), whereas the intracranial pressure remained unchanged (19 (3-45) vs. 11 (5-33) mmHg). No statistical difference was found between the relative changes in the above parameters in survivors compared to non-survivors. CONCLUSION: Although the clinical status did not improve in all patients, both CBF and CMRO2 increased after high-volume plasmapheresis. The alleviation of brain oxygen metabolism by high-volume plasmapheresis may reflect partial removal of neuroinhibitory plasma factors. PMID- 8724029 TI - Cyclosporin A-mediated cholestasis in patients with chronic hepatitis after heart transplantation. AB - Viral chronic hepatitis often occurs in heart transplant recipients receiving cyclosporin. This essential immunosuppressive drug may induce cholestasis. We investigated the effect of treatment with cyclosporin on serum conjugated bile acids in patients with chronic hepatitis developing after heart transplantation. Fifty-nine patients were studied: 17 with chronic hepatitis, 15 heart transplant patients with normal alanine aminotransferase activity, and 27 heart transplant patients with chronic hepatitis, the last two groups receiving cyclosporin. Hepatic biochemical tests and total bile acid concentration were determined on fasting blood samples. The individual glyco- and tauroconjugated bile acids were quantified by high-performance liquid chromatography and direct spectrometry. In patients taking cyclosporin the bilirubin concentration and the alkaline phosphatase activity were increased only when hepatitis was present, in association with a slight increase in cholic acid level (5.13 microM vs. 0.68 microM; P < 0.01). Conjugated lithocholate concentration was dramatically higher when hepatitis and immunosuppression with cyclosporin were associated (1.17 microM vs. 0.03 and 0.04 microM; P < 0.01). Chenodeoxycholate was the main circulating bile acid only in the heart transplant patients treated with cyclosporin but without hepatitis. These results suggest that the mechanisms which explain the cyclosporin-associated modifications of the bile acid pool are different according to the presence or absence of hepatitis. The occurrence of hepatitis in patients on cyclosporin led to an increase in serum lithocholate and primary bile acid concentrations. Further studies are required to assess the effect of ursodeoxycholic acid for this cholestasis. PMID- 8724030 TI - Hepatitis C virus infection in Spanish volunteer blood donors: HCV RNA analysis and liver disease. AB - OBJECTIVE: To evaluate in a large group of volunteer blood donors the prevalence of antibodies to hepatitis C virus (anti-HCV) and the relation of transaminase (ALT) levels and viraemia to liver damage. DESIGN: A prospective study. SETTING: Transfusion Centre of the Autonomous Community of Madrid and the Liver Unit of the Princesa University Hospital. PATIENTS: From a population of 55,587 volunteer blood donors, 160 seropositive cases were further evaluated for virological and histological assessment. METHODS: Anti-HCV was tested by ELISA-2 and RIBA-2 assays. HCV RNA was analysed by nested PCR. Liver biopsies were obtained in 35 volunteer blood donors with abnormal ALT levels. RESULTS: The prevalence of anti HCV detected by ELISA-2 was 0.93%. Serum ALT was abnormal in 61 of the 160 volunteers (38.1%). Of these, RIBA-2 was positive in 96.7% and HCV RNA was detectable in 96.1%. Serum ALT was normal in the remaining 99, 70.7% being RIBA-2 negative and 98.3% HCV RNA negative. The majority of biopsies (85.6%) showed chronic hepatitis. CONCLUSION: This study demonstrates that in blood donors screening for anti-HCV, a positive ELISA-2 test, when associated with abnormal ALT levels, is effective in recognizing subjects with active infection detected by HCV RNA and liver disease. Concerning ELISA-2 positive volunteer blood donors with normal ALT, long-term studies are warranted to elucidate whether they are really infected by HCV. PMID- 8724031 TI - Control of hepatic carbohydrate metabolism and haemodynamics in perfused rat liver by arterial and portal angiotensin II. AB - DESIGN: Angiotensin II (AII; 0.2, 5 and 25 nM) was infused during a single-pass perfusion of a rat liver via both the hepatic artery and the portal vein (portal or arterial AII). Infusion occurred both in the absence and in the presence of the AT1-receptor-antagonist losartan (1 and 10 microM). METABOLISM: Arterial and portal AII increased glucose output and shifted lactate uptake to release. Portal AII was 3 (0.2 nM) and 1.5 times (5 and 25 nM) more effective in increasing hepatic glucose release than similar levels of arterial AII. However, 0.2, 5 and 25 nM of arterially and portally applied AII had a similar level of efficiency in switching lactate uptake to release. The metabolic alterations by arterial and portal AII were, however, strongly inhibited by the addition of 1 microM losartan (an AT1-receptor-antagonist) and completely blocked by the presence of 10 microM losartan. HAEMODYNAMICS: Arterial and portal AII decreased the flow in the ipsilateral vessels to a similar extent, both demonstrating similar kinetics. Medium and high levels of arterial and portal AII caused pronounced flow alterations of the contralateral vessels. The AII-dependent reductions of arterial and portal flow were strongly inhibited by the presence of 1 microM losartan and were stopped by 10 microM of this blocker. RESULTS: The results demonstrate that arterial and portal AII caused alterations in the hepatic metabolism, demonstrating either clear (glucose balance) or no (lactate balance) differences, produced similar reductions of the ipsilateral flow, and pronounced and complex modulations of the contralateral flow. PMID- 8724032 TI - Fulminant hepatitis after carbimazole and propranolol administration. AB - We report the case of a patient who developed jaundice, encephalopathy, a marked increase in serum aminotransferase activity and a decrease in prothrombin and proaccelerin levels, after 6 weeks' treatment with carbimazole and propranolol for hyperthyroidism. The patient ultimately underwent orthotopic liver transplantation. This case strongly suggests that carbimazole may occasionally induce fulminant hepatitis and that careful monitoring of liver enzymes may be useful during the treatment of hyperthyroidism with this drug. PMID- 8724033 TI - Achalasia-like syndrome as the first manifestation in a patient with CREST syndrome. AB - We report a case of oesophageal disease as the first manifestation in a patient with CREST syndrome. A 46-year-old man with achalasia-like syndrome developed CREST syndrome 4 years later. A pneumatic dilatation of the cardia was performed. After pneumatic dilatation the dysphagia and regurgitation disappeared but the patient developed reflux oesophagitis. Four years after diagnosis of oesophageal disease he presented with a clinical picture of CREST syndrome. An acute ileus and constipation developed later. After receiving medical therapy with omeprazole and cisapride the patient is free of oesophageal symptoms and bowel movements are normal. Oesophageal disease is common in patients with limited and diffuse scleroderma, but to our knowledge achalasia-like syndrome has not been previously described as the first manifestation of the systemic disease. PMID- 8724035 TI - Enhanced nuclear factor-kappa B activation induced by tumour necrosis factor alpha in stably tat-transfected cells is associated with the presence of cell surface-bound Tat protein. AB - OBJECTIVE: An enhanced nuclear factor (NF)-kappa B activation in response to tumour necrosis factor (TNF)-alpha has been observed in stably tat-transfected cells. Recent experimental evidence suggests that Tat may autocrinously influence both cellular physiology and HIV-1 long terminal repeat-directed gene expression in Tat-producing cells. Therefore, the possible association of a Tat autocrinous loop with the enhanced NF-kappa B-binding activity induced by TNF-alpha in Tat producing cells was studied by anti-Tat antibody blocking experiments. DESIGN AND METHODS: Permanently tat-transfected Jurkat cells, maintained either in the presence or absence of anti-Tat antibody, were studied for the presence of TNF alpha-induced NF-kappa B-binding activity (quantified by electrophoretic mobility shift assays) and the presence of cell-surface-bound Tat (determined by flow cytometry and confocal microscopy of anti-Tat immunofluorescence-stained cell preparations. RESULTS: The enhanced production of TNF-alpha-induced NF-kappa B binding activity exhibited by tat-transfected Jurkat cells was completely abolished in cell cultures maintained in the presence of anti-Tat antibody, thus indicating that the increased TNF-alpha-induced NF-kappa B binding activity observed in Jurkat-tat cells was dependent on the presence of Tat protein in an antibody-accessible location. In accordance with these findings, immunofluorescence-stained preparations of unfixed tat-transfected Jurkat cells showed the presence of cell-surface-bound Tat protein which was completely absent in cells incubated in the presence of anti-Tat antibodies. CONCLUSIONS: This study demonstrates that the enhanced NF-kappa B activation exhibited by stably tat-transfected cells in response to TNF-alpha, is associated with cell surface interaction of extracellularly released Tat protein. These data add further evidence to the possible relevance of a Tat autocrinous loop in the physiology of Tat-producing cells and suggest that in HIV-1-infected cells Tat is likely to behave as a bifunctional molecule which not only acts from within facilitating NF kappa B recruitment in the viral transcription complex, but may also act from without increasing the availability of activated NF-kappa B. PMID- 8724034 TI - Virological and immunological analysis of a triple combination pilot study with loviride, lamivudine and zidovudine in HIV-1-infected patients. AB - OBJECTIVE: To compare two antiretroviral regiments, loviride plus lamivudine (3TC) plus zidovudine (ZDV) (triple combination) and loviride plus ZDV (double combination) in terms of pharmacokinetic interactions, tolerability, safety, and immunological and virological efficacy. STUDY DESIGN: An open, case-controlled, pharmacokinetic and 24-week continuous treatment pilot study. PATIENTS: Twenty p24 antigen-positive patients, 10 per treatment group, were matched according to p24 antigenaemia less or more than 100 pg, CD4 count less or more than 150 x 10 (6)/l, and gender. Eight out of 10 cases and seven out of 10 controls had received previous antiretroviral therapy. RESULTS: No clinically relevant pharmacokinetic interactions were observed. Both treatment combinations were well tolerated. Median absolute and percentage CD4 count increases above baseline were more pronounced in the triple combination arm than in the double combination arm. Median p24-antigen and plasma viraemia level decreases below baseline were more pronounced in the triple combination arm. The M(184)I/V mutation was detected in all plasma samples of triple combination patients examined at week 12. Mutations conferring resistance to loviride and ZDV were found in a significant subset of patients in both treatment arms. CONCLUSIONS: Both combination regimens have an excellent safety/tolerability profile, but a higher level of in vivo efficacy is achieved by the triple combination, despite genotypic changes conferring resistance to one or all of these agents. The conclusions drawn are limited by small population size and the heterogenous pretreatment history. However, they support the validity of and strongly encourage a rationally designed multidrug combination approach to HIV therapy. PMID- 8724036 TI - Human brain N-methyl-D-aspartate receptors regulating noradrenaline release are positively modulated by HIV-1 coat protein gp120. AB - OBJECTIVE: To investigate the effect of HIV-1 gp120 on the function of glutamate receptors of the N-methyl-D-aspartate (NMDA) type in the human brain. DESIGN: The monitoring of neurotransmitter release from superfused isolated nerve endings is widely recognized as a technique appropriate for the study of neurotransmitter release and to attribute a precise localization to the site(s) of action of drugs able to modulate release. METHODS: Synaptosomes (pinched-off nerve endings) were prepared from fresh human brain tissue samples removed during neurosurgery, labelled with [3H]-noradrenaline and superfused at a rate of 0.5 ml/min with NMDA in the presence of gp41, gp160, gp120 or the V3 loop, with or without NMDA receptor antagonists. Fractions of superfusate were collected and measured for radioactivity. RESULTS: NMDA elicited a glycine-sensitive release of [3H] noradrenaline from human brain synaptosomes. HIV-1 gp120 potentiated the NMDA (1 mM)-evoked [3H]-noradrenaline release (maximal effect approximately 110% at 1 nM). The release elicited by NMDA plus gp120 was prevented by the classical NMDA receptor antagonists dizocilpine or 7-chlorokynurenic acid, as well as by memantine. The potentiation by gp120 of the NMDA-evoked [3H]-noradrenaline release was mimicked by gp160 but not by gp41. The effect of gp120 was retained by the V3 loop. Finally, gp120 reversed (1 nM) and surmounted (10nM) the antagonism by 10 microM 7-chlorokynurenate of the NMDA-evoked [3H]-noradrenaline release. CONCLUSION: gp 120 binds directly through the V3 loop at noradrenergic axon terminals in human brain neocortex and may alter the function of presynaptic NMDA receptors mediating regulation of noradrenaline release. PMID- 8724037 TI - Pentoxifylline inhibits acute HIV-1 replication in human T cells by a mechanism not involving inhibition of tumour necrosis factor synthesis or nuclear factor kappa B activation. AB - OBJECTIVE AND DESIGN: To study the in vitro activity of pentoxifylline (PTX), which may be of benefit in AIDS, on cell proliferation, tumour necrosis factor (TNF)-alpha, interferon (IFN)-gamma (a type 1 cytokine) and interleukin (IL)-10 (a type 2 cytokine) production, viral replication and CD4+ depletion in acutely HIV-1-infected human T cells. METHODS: T cells were stimulated with anti-CD3 antibody or phytohaemagglutinin (PHA) and infected with HIV-1 in presence or absence of PTX. Cell proliferation, CD4+ cell number, nuclear factor (NF)-kappa B activation, p24 antigen release or lymphokine content of the supernatants were evaluated by [3H]-thymidine incorporation, cytofluorimetry, electrophoretic mobility shift assays and specific enzyme-linked immunosorbent assay, respectively. RESULTS: In HIV-1-infected T cells, PTX inhibited cell proliferation and p24 release and prevented CD4+ depletion associated with HIV replication. Moreover, PTX reduced TNF-alpha, IFN-gamma and IL-10 production and NF-kappa B activation. PTX inhibited with similar potency IFN-gamma, TNF-alpha and cell proliferation. However, the inhibition of p24 release and specially of IL-10 production required significantly lower doses of PTX. Exogenous addition of IL-2 or TNF-alpha in presence of PTX restore T-cell proliferation and NF-kappa B activation respectively, but did not affect p24 inhibition. CONCLUSIONS: Our data suggest that the inhibitory effect of PTX on HIV replication cannot be satisfactorily explained by the inhibition of NF-kappa B or TNF-alpha. Moreover, PTX cannot be primarily considered as a TNF-alpha inhibitor and has several immunomodulatory and antiviral properties which could be of benefit against HIV-1 at various levels. PMID- 8724038 TI - Inhibition of fungicidal activity of polymorphonuclear leukocytes from HIV infected patients by interleukin (IL)-4 and IL-10. AB - OBJECTIVE: To investigate the effect of human recombinant interleukin (hrIL)-4 or hrIL-10 on the functional status of polymorphonuclear leukocytes (PMNL) from normal subjects and HIV-infected patients. DESIGN: In an in vitro system we studied the effect of hrIL-4 or hrIL-10 on phagocytosis, fungicidal activity and superoxide anion production by PMNL. METHODS: PMNL were treated in vitro with hrIL-4 or hrIL-10 or their combination for 6 h and then candidacidal activity was evaluated in a colony-forming unit inhibition assay. Superoxide anion generation by PMNL was measured in the presence or absence of preopsonized zymosan or Candida albicans. RESULTS: Treatment in vitro with hrIL-4 or hrIL-10 of PMNL for 6 h was able to impair candidacidal activity of neutrophils in both normal or HIV infected patients. The inhibitory effect was time- and dose-dependent and was more evident in PMNL from HIV-infected subjects, and reflected in these latter cells a decrease of superoxide anion generation. The impairment of candidacidal activity in PMNL from HIV-infected patients was accompanied by survival of the yeasts shown by budding formation into phagosomic organelles of cytokine-treated PMNL. CONCLUSIONS: Our data highlight new biological effects of IL-4 and IL-10 evidenced by suppressed effector function of neutrophils; this phenomenon is emphasized in HIV-infected patients suggesting a role for these cytokines in mediating increased susceptibility to microbial infection during AIDS progression. PMID- 8724039 TI - A 24-week open-label phase I/II evaluation of the HIV protease inhibitor MK-639 (indinavir). AB - OBJECTIVE: To investigate the safety, pharmacokinetics, and activity of the orally bio-available protease inhibitor MK-639. DESIGN: An open-label Phase I/II trial of medically stable subjects with screening CD4 lymphocyte counts < or = 300 x 10(6)/I and > or = 20,000 HIV RNA copies/ml. Pharmacokinetics were performed at days 1 and 15. In order to better understand the relationships between drug exposure, baseline activity markers, and their changes during the study, mathematical modeling was performed using the traditional sigmoid-Emax relationship of pharmacologic effect and first order inhomogeneous differential equations for a two compartment system. RESULTS: The five men enrolled had extensive prior nucleoside therapy (mean, 32.6 +/- 25.6 months), a low mean CD4 lymphocyte cell count (CD4 count, 66.1 +/- 61 x 10(6)/I and CD4 percentage, 4.4 +/- 3.1%), high soluble tumor necrosis factor-alpha type II (sTNFII) receptor concentration (6.23 +/- 2.76 ng/ml) and high viral load (5.13 +/- 0.46 log10 RNA copies/ ml; geometric mean, 133,941 copies/ml). The drug was well tolerated at a dose of 600 mg every 6 h. The steady state concentrations Cmax and Cmin were 4.94 +/- 2.16 microM and 0.28 +/-0.1 microM, respectively, which are approximately equal to 50 and 3 times the 95% inhibitory concentration (IC95) for clinical isolates, respectively. The mean increase in CD4 cell count was 143 x 10(6)/ (217% increase ), the mean increase in CD4 percentage was 5.2 percentage points (118%), mean decrease in HIV RNA was 1.55 log10 RNA copies/ml (a geometric mean difference of 130,120 copies/ml or 97% decrease) with a slow upward drift on continued therapy to a mean 0.64 log10 RNA copies/ml decrease by week 24 (a geometric mean difference of 103,084 copies/ml or 77% decrease), and a mean decrease in sTNFII receptors of 2.78 ng/ml (45% decrease). The mean CD4 counts per week as a function of the starting CD4 counts fit a sigmoid-Emax relationship (r2 = 0.998, P < 0.0001) with the return of CD4 cells being strongly related to the number of CD4 cells at baseline. Drug exposure as measured by either the total exposure (area under the concentration/time curve) or as the Cmin gave similar significant relationships to the fractional inhibition of HIV generation (r2 = 0.999, P < 0.0001, and r2 = 0.996, P < 0.0001, respectively). CONCLUSIONS: MK-639 appears to have significant dose-related antiviral activity and is well tolerated. PMID- 8724040 TI - Influence of risk group and zidovudine therapy on the development of HIV encephalitis and cognitive impairment in AIDS patients. AB - OBJECTIVE: To determine the associations between HIV encephalitis and other central nervous system (CNS) pathology, viral burden, cognitive impairment, zidovudine therapy and risk group in AIDS patients. DESIGN: Planned autopsy study in AIDS patients evaluated prospectively for numerous clinical parameters. SETTING: Regional academic centre for clinical care and pathology examination of patients with HIV infection. PATIENTS: Edinburgh cohort of HIV-positive patients prospectively assessed for cognitive impairment, immunosuppression and clinical course. Unbiased series of consecutive autopsies in 27 homosexual men and 39 drug using patients with AIDS. INTERVENTIONS: Zidovudine therapy monitored in all patients. MAIN OUTCOME MEASURES: Determination of CNS viral burden and pathology including immunocytochemically confirmed HIV encephalitis in injecting drug users (IDU) versus homosexual AIDS patients with known CD4 counts and cognitive function. RESULTS: HIV encephalitis was present in 59% of IDU and 15% of homosexuals: 88% of patients with encephalitis had displayed cognitive impairment. HIV encephalitis was strongly associated with a high viral load and HIV p24 immunopositivity. Opportunistic infections and lymphomas were more common in homosexuals (63%) than in IDU (31%) and were associated with the degree of immunosuppression before death. Within both groups, prolonged zidovudine treatment was associated with a lower incidence of HIV encephalitis. CONCLUSIONS: This study documents two separate CNS outcomes in AIDS patients in that HIV encephalitis occurs independently of opportunistic infections and lymphomas and shows different associations with risk group, immunosuppression and antiviral treatment before death. PMID- 8724041 TI - Intracellular glutathione in the peripheral blood cells of HIV-infected patients: failure to show a deficiency. AB - OBJECTIVE: To determine whether HIV-infected patients have a deficiency of intracellular glutathione (GSH) in peripheral blood mononuclear cells (PBMC) and erythrocytes. DESIGN: Initial experiments determining the stability of intracellular GSH preceded the measurement of GSH levels in 33 HIV-positive patients and 40 control subjects within 1 h of isolation of their blood cells. In addition, the susceptibility of erythrocytes to dapsone hydroxylamine-induced methaemoglobinaemia was evaluated. METHODS: GSH levels were determined by an high performance liquid chromatography method utilizing a fluorescent probe, monobromobimane. The bimane-GSH adduct formed in PBMC was also characterized by mass spectrometry. Methaemoglobin formation on exposure to dapsone hydroxylamine was determined spectrophotometrically. RESULTS: GSH levels remained stable for only 1 h after cell isolation, thereafter showing a decrease of 20 and 60% at 4 and 24H, respectively, There was no difference in the GSH levels in PBMC and erythrocytes of the HIV-positive patients compared with controls. The GSH levels were not related to the disease stage or to CD4+ cell counts. There was no difference in GSH levels in PBMC taken from trimethoprim-sulphamethoxazole hypersensitive and non-hypersensitive patients. Methaemoglobinaemia on exposure of erythrocytes to dapsone hydroxylamine was concentration-dependent, but there was no significant difference between patients and controls. CONCLUSION: In contrast to previous studies, no deficiency of intracellular GSH in the PBMC and erythrocytes of HIV-infected patients was found. The discrepancy between studies may be methodological reflecting the instability of GSH, which requires prompt sample analysis. PMID- 8724042 TI - Reasons for failure of prevention of Toxoplasma encephalitis. Swiss HIV Cohort Study. AB - OBJECTIVE: To study whether absence of prophylaxis is a risk factor for cerebral toxoplasmosis, and to determine the reasons for absence of prophylaxis among AIDS patients diagnosed with Toxoplasma encephalitis (TE). DESIGN: Retrospective chart review and matched case-control study. PATIENTS: Patients (104 first episodes and 26 relapses of TE) were registered in the Swiss HIV Cohort Study from three centres from July 1992 to December 1994; 91 matched controls were included for 52 patients with a first episode, and 17 matched controls for 17 patients with relapse of TE. RESULTS: Prophylaxis was prescribed to 17 patients (16%) with a first episode and 19 patients (73%) with a relapse of TE. Reasons for the absence of prophylaxis included patient refusal (25%), non-proposal by physicians (17%), and drug intolerance (17%). Reduced absorption due to non-compliance, diarrhoea or vomiting was identified among 12 patients with a first episode and 14 patients with a relapse of TE. Absence of prescription of prophylaxis was associated with a 10-fold increased risk of a first episode of TE (odds ratio, 9.8; 95% confidence interval, 2.7-35.4) in the matched case-control study. CONCLUSIONS: TE continues to occur among patients not receiving prophylaxis. At least one-half of the cases may be prevented with better motivation of physicians and increased compliance of patients. PMID- 8724043 TI - Randomized cross-over comparison of liposomal daunorubicin versus observation for early Kaposi's sarcoma. AB - OBJECTIVES: To evaluate single-agent liposomal daunorubicin chemotherapy in the management of early HIV-related Kaposi's sarcoma (KS). DESIGN: Randomized cross over comparison of liposomal daunorubicin versus observation. SETTING: Study conducted at single site in tertiary referral HIV unit. PATIENTS: Twenty-nine HIV seropositive men with < 20 cutaneous KS, no visceral involvement and CD4 cell counts < 400 x 10(6)/I were randomized. Adequate haematological, hepatic and renal function was required for entry. A left ventricular ejection fraction of > 45% was necessary for eligibility. INTERVENTIONS: Patients were randomized to 12 weeks observation or 12 weeks of liposomal daunorubicin 40 mg/m2 every 2 weeks. After 12 weeks, or at disease progression, patients were crossed over to receive the alternative arm. MAIN OUTCOME MEASURES: Disease evaluation was according to AIDS Clinical Trials Group criteria for response assessment and toxicity was recorded using the World Health Organization standardized grading. RESULTS: Response rate to initial liposomal daunorubicin was six out of 15 (40%) and none experienced a spontaneous response during the observation arm. Six patients (40%) randomized to the initial chemotherapy arm progressed during chemotherapy, while 10(72%) in the observation arm progressed. Neutropenia was the main toxicity associated with liposomal daunorubicin and was documented following 20 out of 139(14%) treatment cycles. CONCLUSIONS: Liposomal daunorubicin is a well tolerated and efficacious treatment for early KS; however, the duration of response is brief. PMID- 8724044 TI - Nosocomial outbreak of tuberculosis in an outpatient HIV treatment room. AB - OBJECTIVE: To investigate a possible outbreak of tuberculosis in an outpatient HIV treatment facility in Sydney, Australia. DESIGN: Following the diagnosis of pulmonary tuberculosis in an attendee, a prospective screening program was instituted to investigate the potential outbreak. METHODS: Screening of 89 potentially exposed patients included chest radiographs (n = 89), and sputum examination (n = 37) over a period of 23 weeks. RESULTS: No cases of tuberculosis were detected by the screening program. However, three (3.4%) of this cohort developed pulmonary tuberculosis between 8 and 10 weeks following diagnosis of the index case. The incidence of active tuberculosis during the following-up period (median, 7.4 months) was 5.3 per 100 person years and represents the lower limit of possible tuberculous infection, as both latent infection, and undiagnosed tuberculosis among those who died could not be excluded. Mycobacterium tuberculosis strains isolated from the index case and three subsequent cases were found to be identical by DNA typing. CONCLUSION: Nosocomial transmission of tuberculosis in an outpatient treatment setting has been demonstrated. The risk of nosocomial transmission of tuberculosis is significant in institutions caring for HIV-infected patients even in countries with a low prevalence of tuberculosis infection, and highlights the importance of adherence to tuberculosis control guidelines. PMID- 8724045 TI - Rapid increase in HIV-related tuberculosis, Chiang Rai, Thailand, 1990-1994. AB - OBJECTIVE: Chiang Rai, the northernmost province of Thailand, has experienced an explosive HIV epidemic since 1989. This study assessed the impact of HIV infection on tuberculosis (TB) in the area. METHODS: We analyzed the incidence of reported TB in the province from 1982 through 1993 and TB registry data at Chiang Rai Hospital from 1985 through 1994. RESULTS: Following a steady decline in reported TB from 1982 through 1991, the incidence of TB increased sharply after 1991. TB registry data from Chiang Rai Hospital, which began confidential HIV testing in October 1989, indicated a steady and rapid increase in the number and proportion of HIV-seropositive TB patients from four (1.5% of all TB patients) in 1990 to 207 (45.5%) in 1994 (P < 0.001). Compared with HIV-negative TB patients, HIV-positive TB patients were more likely to be men, aged 20-39 years and have extrapulmonary TB (P < 0.001). Treatment completion rates were similar. Twelve months after beginning TB treatment, HIV-positive TB patients had a mortality rate of 68.6% [95% confidence interval (Cl), 62.7-74.3] compared with 10.0% (95% Cl, 8.3-12.1%) in HIV-negative patients (P < 0.001). CONCLUSION: Thailand and other Asian countries where HIV is spreading rapidly must promptly address the dual epidemic of TB and HIV in order to reduce preventable morbidity and mortality. PMID- 8724046 TI - Sexual mixing patterns in Uganda: small-time urban/rural traders. AB - OBJECTIVE: To document the extent to which rural and urban-based male traders have sexual contacts away from their place of residence. METHODS: Fifty-one traders (46 who travel by bicycle and five others) kept daily diaries of all their journeys and sexual contacts for a total of 584 person-weeks. Twenty-five were resident in a trading town where HIV prevalence was about 40% and 26 lived up to 25 km away in rural areas where HIV prevalence was approximately 8%. RESULTS: A total of 2147 return trips were made (mean, 3.7 per week). Eighty per cent were between the trading town and the surrounding rural area. A total of 1377 sexual contacts were recorded (mean, 2.3 per week); 95% of the contacts of urban-based men occurred in the town, 3% in other urban areas and 2% in a rural area. For rural-based men 82% of sexual contacts took place in their home village, 14% in a neighbouring village, 2% in the trading town and 3% in other urban centres. CONCLUSION: Despite considerable economic interaction there is very little sexual mixing between the town and surrounding rural areas. This may explain why the high HIV prevalence found in some trading towns in Africa has not diffused out to rural areas. PMID- 8724047 TI - A controlled HIV/AIDS-related health education programme in Managua, Nicaragua. AB - OBJECTIVES: To evaluate the impact of a community-wide intervention to increase HIV/AIDS-related knowledge, change attitudes and increase safer-sex practices in Managua, Nicaragua. DESIGN AND SETTINGS: Household-based health education intervention trial comprising a knowledge, attitudes and practices (KAP) survey at baseline, a health education intervention and a follow-up KAP survey. Four neighbourhoods were included, two received the intervention, and the other two served as controls. Randomly selected residents aged 15-45 years were interviewed at baseline (n = 2160) and follow-up (n = 2271) using an identical questionnaire. The intervention consisted of a health education campaign that emphasized HIV transmission and condom use. OUTCOME AND ANALYSIS: Knowledge levels regarding transmission and prevention of HIV infection, self-reported use of condoms, levels of worries about HIV/AIDS and perceptions of personal risk of HIV infection. Comparisons between baseline and follow-up employed chi 2 tests with continuity correction. The influence of the intervention was examined in multivariate logistic models including an appropriate interaction term. RESULTS: Intervention and control samples were comparable with regard to sex, age, and age at first intercourse. Significantly less intervention residents had formal education (P < 0.001). At baseline, outcome variables were generally similar in control and intervention samples. Condom use increased from 9 to 16% (P = 0.003) among intervention women, but only from 9 to 11% (P = 0.5) in control women (test for interaction, P = 0.08). Among men, increases were from 31 to 41% (P < 0.001) and from 30 to 37% (P = 0.06), respectively (test for interaction, P = 0.3). Levels of worries about HIV/AIDS decreased in all groups, but perception of individual risk increased only among intervention women (test for interaction, P = 0.02). CONCLUSIONS: This household-targeted health education intervention appears to have had some effect; however, sustained efforts are needed further to improve levels of knowledge and to increase condom use in Managua. PMID- 8724048 TI - Human herpesvirus-8 in AIDS-related and unrelated lymphomas. PMID- 8724049 TI - Interaction between lamivudine (3TC) and other nucleoside analogues for intracellular phosphorylation. PMID- 8724050 TI - Identification of HIV-1 viral RNA in intestinal tissue from patients with early and late HIV infection. PMID- 8724051 TI - T-cell receptor V beta-specific expansions in children from HIV-infected mothers. PMID- 8724052 TI - Hospital-based surveillance and emerging patterns of HIV transmission. Groupe d'Epidemiologie Clinique du SIDA en Aquitaine (GECSA) PMID- 8724053 TI - Where's the virus? Monitoring current HIV transmission patterns. PMID- 8724054 TI - Origin of hyperamylasaemia in HIV-infected patients. PMID- 8724055 TI - HIV infection among antenatal women in Zambia, 1990-1993. PMID- 8724056 TI - Factors related to inconsistent condom use with commercial sex workers in northern Thailand. PMID- 8724057 TI - The efficacy of nonoxynol-9 from an in vitro point of view. PMID- 8724058 TI - A hundred years of vaccination: the legacy of Louis Pasteur. PMID- 8724059 TI - Varicella vaccine and health care workers. PMID- 8724060 TI - Trends in hospitalizations for diarrhea in United States children from 1979 through 1992: estimates of the morbidity associated with rotavirus. AB - OBJECTIVES: To examine trends in the hospitalizations of children for diarrheal disease in the U.S. and to provide estimates for the burden of disease associated with rotavirus diarrhea. METHODS: Data for diarrheal hospitalizations among U.S. children ages 1 month through 4 years were compiled from the National Hospital Discharge Survey for the years 1979 through 1992. Between 1979 and 1992, 12% of all hospitalizations of U.S. children 1 month through 4 years of age had an International Classification of Diseases code for diarrhea listed in one of the top three positions on the discharge diagnosis. RESULTS: The annual rate of diarrheal hospitalizations, 97 per 10 000 persons (average, 185 742 per year), did not change substantially during the 14-year study period and accounted annually for 724 394 inpatient days (3.9 days per hospitalization). For most diarrheal hospitalizations (75.9%) no causative agent was specified in the National Hospital Discharge Survey records; of the remaining 24.8%, viruses were most commonly reported (19.3%), followed by bacteria (5.1%) and parasites (0.7%). The proportion of hospitalizations associated with viral diarrheas rose from 13% to 27% during the 14-year study period, whereas the proportion of hospitalizations for noninfectious diarrhea declined from 79% to 60%. Every year the number of hospitalizations peaked from November through April, the "winter" months, among children ages 4 through 35 months; this peak began in the West during November and December and reached the Northeast by March. CONCLUSIONS: Diarrhea continues to be a common cause of hospitalization among children in the United States and the winter seasonality estimated to be caused in large part by rotavirus would be expected to decrease if rotavirus vaccines currently being developed were introduced. Our analysis of temporal trends in diarrheal hospitalizations provides a unique surrogate with which to estimate the disease burden associated with rotavirus diarrhea. PMID- 8724061 TI - Adverse reactions in children during long-term antimicrobial therapy. AB - BACKGROUND: It is difficult to obtain reliable date on the rate of adverse reactions caused by drugs in general use. Yet it would be important to compile data on adverse reactions to long-term antimicrobial therapy. METHODS: A sample of 1607 girls and 218 boys from 16 409 children younger than 16 years who had received long term antimicrobial therapy for recurrent urinary tract infections during 1976 to 1985 was analyzed with regard to adverse reactions. RESULTS: Altogether 5066 courses of treatment were given to female patients and 607 to male patients. Adverse reactions were reported in 589 courses of the 5673 (10.4%), and 463 courses (8.2%) were discontinued because of adverse reactions. None of the patients had serious life-threatening reactions, and none of those receiving nitrofurantoin had pulmonary problems. The most common adverse reactions associated with the use of nitrofurantoin were nausea and vomiting (rate, 4.4/100 person years at risk; 95% confidence interval, 3.4 to 5.4), whereas sulfonamides caused most commonly allergic skin reactions (rate, 4.6; 95% confidence interval, 3.2 to 6.5). Patients younger than the age of 2 years receiving nitrofurantoin had adverse reactions more often than those who received sulfonamides, but in the age group 2 to 15 years sulfonamides caused adverse reactions leading to discontinuation of treatment more often than did nitrofurantoin of treatment the adverse reactions occurred during the first 6 months of treatment. CONCLUSIONS: We found nitrofurantoin and sulfonamides to be safe drugs for use in long term preventive antimicrobial therapy. PMID- 8724062 TI - Efficacy of ceftibuten compared with amoxicillin for otitis media with effusion in infants and children. AB - OBJECTIVES: This trial compared the efficacy of ceftibuten with that of amoxicillin in resolving otitis media with effusion. METHODS: Two hundred ten children with otitis media with effusion were randomly assigned to receive either ceftibuten (9 mg/kg/day in one daily dose) or amoxicillin (40 mg/kg/day divided into 3 daily doses) for 14 days. Outcome was assessed at 2 and 4 weeks in all subjects; those without middle ear effusion at the 4-week visit were examined at 8, 12 and 16 weeks. Middle ear status was determined by pneumatic otoscopy and by an algorithm combining pneumatic otoscopy with tympanometry. RESULTS: The percentages of subjects who were effusion-free in the ceftibuten and amoxicillin groups at 2 weeks by otoscopy were 29.8 and 27.2%, respectively (P = 0.59), and by the algorithm, 23.4 and 20.4%, respectively (P = 0.47). In children who were effusion-free at 2 weeks, recurrence of effusion by 16 weeks was noted in approximately 60% of the ceftibuten group and 67% of the amoxicillin group. No medication side effects were reported by 88% of subjects in the ceftibuten group and by 93% of subjects in the amoxicillin group. We found no significant differences between the ceftibuten and amoxicillin groups with respect to resolution of middle ear effusion, rate of recurrence or side effects. CONCLUSIONS: Amoxicillin remains the drug of first choice for treatment of otitis media with effusion when treatment is deemed advisable, but ceftibuten may be an alternative drug in selected situations. PMID- 8724063 TI - Incidence of Vibrio cholerae O1 diarrhea in children at the onset of cholera epidemic in periurban Lima, Peru. AB - OBJECTIVE: To determine the incidence of Vibrio cholerae O1-associated diarrhea in children during the onset of the 1991 cholera epidemic in Peru. METHODS: Stool cultures were obtained from children (mean age, 26 months) participating in a prospective community-based study of diarrhea in a periurban community in Lima between February and May, 1991. RESULTS: Of the 409 diarrheal episodes cultured V. cholerae O1 was isolated in 14 (3.4%) episodes. This represented an incidence of 0.11 episode per child year, higher than previously reported rates in children from endemic cholera areas. Most cases were mild; only 1 case required hospitalization. CONCLUSIONS: Our study indicates that from the beginning of this epidemic, V. cholerae O1 caused diarrhea in children as well as adults and should therefore be considered as one of the possible pathogens when children from a cholera-affected area develop diarrhea. PMID- 8724064 TI - Immunoglobulin allotypes and genetic susceptibility to invasive Haemophilus influenzae type b and Staphylococcus aureus infections in South African children. AB - OBJECTIVE: The purpose of this study was to determine whether the G2m(n), G1m(f) and Km(3) immunoglobulin allotypes have any association with susceptibility to invasive Haemophilus influenzae type b (Hib) and Staphylococcus aureus (S. aureus) infections in children. METHODS: Direct enzyme-linked immunosorbent assays with commercially available monoclonal antibodies were established to quantitate G2m(n) and G1m(f) allotypes. A qualitative enzyme-linked immunosorbent assay with polyclonal rabbit anti-Km(3) antibody was established for Km(3) determination. RESULTS: The G2m(n) marker occurred in 34.4% of the mixed ancestry population and 2.9% of the Black population. There was a significantly decreased frequency of the G2m(n) allotype in mixed ancestry children with Hib meningitis (8.5%) and Hib osteomyelitis/septic arthritis and a decreased frequency of Km(3) in black and mixed ancestry children with Hib meningitis. The frequency of G2m(n), G1m(f) and Km(3) allotypes in patients with S. aureus osteomyelitis/septic arthritis were not significantly different from normal population frequency. CONCLUSIONS: This study shows a clear association between the absence of the G2m(n) allotype in mixed ancestry children and susceptibility to invasive infections caused by H. influenzae and an association between the absence of Km(3) and Hib meningitis in both black and mixed ancestry children. PMID- 8724066 TI - Pneumococcal disease among children in a rural area of west Africa. AB - BACKGROUND: The pneumococcus is a frequent cause of pneumonia and other serious infections among young children in developing countries. Defining the pattern of pneumococcal infection in these countries is important so that, with the advent of pneumococcal conjugate vaccines, rational vaccination policies can be developed. METHODS: Children younger than 5 years of age who attended clinics in a rural area of The Gambia, West Africa, were screened by assistants during a 2 year period. Children with predefined features suggestive of a diagnosis of pneumonia, meningitis or septicemia were referred to the Medical Research Council Field Station at Basse for investigation. RESULTS: Of 2898 children investigated 103 cases of invasive pneumococcal disease (70 definite and 33 probable) were identified, suggesting that the incidence of this infection in the study community is at least 554/100,000/year in children younger than 1 year of age and 240/100,000/year in those younger than 5 years, rates many times higher than those found in industrialized societies. The mean age of presentation was 15 months; more boys than girls were affected. Cases of pneumonia were encountered 8 times more frequently than those of meningitis. Antibiotic resistance was rarely found and cases of pneumonia, but not meningitis, responded well to treatment. Case-fatality rates in children with pneumonia and meningitis were 1 and 55%, respectively. The most prevalent pneumococcal serotypes were types 6, 14, 19, 1 and 5. CONCLUSION: About 60% of invasive pneumococcal infection in children in this community could potentially be prevented by a nine-valent pneumococcal conjugate vaccine (types 1, 4, 5, 6B, 9, 14, 18, 19F and 23) given at the ages of 2, 3 and 4 months. PMID- 8724065 TI - Immunologic priming of young children by pneumococcal glycoprotein conjugate, but not polysaccharide, vaccines. AB - BACKGROUND: Streptococcus pneumoniae is the most common cause of invasive bacterial disease and otitis media in infants and young children. Licensed pneumococcal polysaccharide vaccines are not reliably immunogenic in children younger than 2 years of age; therefore pneumococcal glycoprotein conjugate vaccines are currently being evaluated for safety, immunogenicity and efficacy in various age groups. METHODS: During a 12-month period we determined the kinetics of pneumococcal IgG antibody in 60 children who received primary immunization with one dose of bivalent (serotypes 6A and 23F) pneumococcal polysaccharide CRM197 vaccines at 18 to 30 months of age. To assess immunologic priming a subgroup of 20 subjects received secondary immunization with pneumococcal polysaccharide vaccine, including serotypes 6B and 23F, at 11 to 20 months after primary immunization. Pneumococcal-specific IgG subclass distributions were also evaluated in the subgroup. RESULTS: In the 12 months after primary immunization with glycoprotein conjugate vaccine, geometric mean pneumococcal IgG antibody concentrations to 6B and 23F serotypes remained stable. Pneumococcal polysaccharide vaccine induced a greater anamnestic response in children primed with glycoprotein conjugate vaccines (13- to 40-fold increases to geometric mean concentrations of 6 to 30 micrograms/ml for type 23F), than in those primed with polysaccharide (2- to 4-fold increases). A greater IgG response to pneumococcal serotype 23F than to 6B was observed with both primary and secondary immunization. The serotype-specific pneumococcal IgG antibody response was virtually restricted to the IgG1 subclass after primary immunization, but secondary immunization elicited antibodies of IgG1 and IgG2 subclasses. CONCLUSIONS: These glycoprotein conjugate vaccines appear to prime for anamnestic IgG antibody responses to subsequent immunization with polysaccharide vaccine, suggesting that the polysaccharide-CRM197 vaccine effectively induces a predominantly T cell-dependent immune response. The greater IgG response to 23F than to 6B indicates that pneumococcal serotype is a major determinant of immunogenicity of pneumococcal glycoprotein conjugate vaccines. PMID- 8724067 TI - Clinical spectrum of human immunodeficiency virus disease in children in a west African city. Project RETRO-CI. AB - OBJECTIVES: To determine the prevalence of HIV infection in children and to compare diagnostic syndromes and outcomes in HIV-positive and HIV-negative children. METHODS: Consecutive children hospitalized in Abidjan's three university hospitals were examined, tested for HIV infection and followed to discharge. Admission or discharge diagnoses and outcome (survived or died) were compared in HIV-positive and HIV-negative children. RESULTS: The prevalence of HIV infection in the 4480 children hospitalized for the first time was 8.2%; the highest age-specific rate (11.2%) was in children ages 15 to 23 months. Six clinical syndromes accounted for more than 80% of admissions in HIV-positive and negative children (all ages combined): respiratory infection; malnutrition; malaria; anemia; diarrhea; and meningitis. The dominant syndromic diagnoses in HIV-positive children were respiratory infection (26.1%) and malnutrition (25.8%); in HIV-negative children they were malaria (30.4%) and respiratory infection (19.1%). The overall mortality rate in HIV-positive children was 20.8%, compared with 8.7% in HIV-negative children (relative risk, 2.4; 95% confidence interval, 1.9 to 3.1); the highest death rate (28.1%) was in children younger than 15 months. CONCLUSIONS: Clinical syndromes associated with HIV infection in African children are difficult to recognize without access to HIV serology. Respiratory infection and malnutrition were the dominant clinical syndromes in HIV-positive children in Abidjan. Greater overlap exists between the clinical presentations of HIV-associated disease and other common health problems in African children than in adults. PMID- 8724068 TI - Clinical manifestation and outcome of tuberculosis in children with acquired immunodeficiency syndrome. AB - BACKGROUND: Atypical clinical manifestations and rapid progression of tuberculous disease (TB) are well-recognized in adults with the acquired immunodeficiency syndrome (AIDS). There are few reports of children with AIDS and TB. We report the manifestations, clinical course and outcome of 12 pediatric patients with AIDS and TB. METHODS: The charts of all children admitted to our institution, from 1989 through 1994, with the diagnoses of AIDS and culture-proved TB were reviewed. RESULTS: Twelve children between the ages of 2 months and 13 years fit the criteria. The mean time between the diagnosis of AIDS and TB was 20 months. The most frequent presenting symptoms were fever (75%) and tachypnea (33%). All had negative Mantoux tests (5 tuberculin units of purified protein derivative). Extrapulmonary TB was present in 3 (25%). A source case was identified for 4 (33%). Previous pulmonary disease was present in 7 (58%). Chest roentgenograms were abnormal in 11 (91%), with diffuse interstitial infiltration the most common finding. Susceptibility tests were performed on 10 strains, 3 of which were resistant to 1 or more antituberculosis drugs. Three patients (25%) died of TB, 1 of whom was appropriately treated with antituberculosis drugs but had a strain resistant to isoniazid and rifampin. CONCLUSION: Children with AIDS and TB most frequently present with atypical manifestations of TB. A high index of suspicion is needed to correctly diagnose TB in this group of children. Early diagnosis is important because most respond well when treated appropriately. PMID- 8724069 TI - Bronchiolitis obliterans organizing pneumonia in a child with acquired immunodeficiency syndrome. PMID- 8724070 TI - Infections complicating lawn mower injuries in children. PMID- 8724072 TI - Varicella disease and transmission in pediatric house officers. PMID- 8724071 TI - Rapid oral desensitization to trimethoprim-sulfamethoxazole in infants and children. AB - BACKGROUND: Although trimethoprim-sulfamethoxazole is the preferred chemoprophylaxis against Pneumocystis carinii pneumonia, there are frequent IgE mediated reactions among children infected with the human immunodeficiency virus (HIV). Oral desensitization allows more patients to receive chemoprophylaxis, but it has been studied in only a limited number of children. METHODS: We desensitized five children infected with the HIV using a rapid, 4-h oral protocol. RESULTS: Three children (including two infants) successfully completed desensitization and started maintenance therapy, but the other two experienced reactions that precluded further administration of trimethoprim-sulfamethoxazole. CONCLUSIONS: We conclude that a rapid, oral trimethoprim-sulfamethoxazole desensitization protocol is safe and, in some instances, effective among HIV infected children and infants with a history of non-life-threatening, IgE mediated reactions to trimethoprim-sulfamethoxazole. PMID- 8724073 TI - Pulmonary tuberculosis after lung-liver transplantation from cystic fibrosis. PMID- 8724074 TI - Cutaneous aspergillosis in a neonate. PMID- 8724075 TI - Prolonged thalidomide therapy for human immunodeficiency virus-associated recurrent severe esophageal and oral aphthous ulcers. PMID- 8724076 TI - Fatal intrauterine infection associated with penicillin-resistant Streptococcus pneumoniae. PMID- 8724077 TI - Group B streptococcal sepsis and meningitis complicated with severe sensorineural hearing loss in a fourteen-year-old boy. PMID- 8724078 TI - Alice in Wonderland syndrome caused by coxsackievirus B1. PMID- 8724079 TI - Herpes zoster ophthalmicus with delayed contralateral hemiplegia. PMID- 8724080 TI - The school child with human immunodeficiency virus. PMID- 8724081 TI - Recommendations for hospital visitation by recipients of varicella vaccine. PMID- 8724082 TI - Morphometric analysis of the lumbosacral nerve roots and dorsal root ganglia by magnetic resonance imaging. AB - STUDY DESIGN: Lumbosacral nerve roots and dorsal root ganglia in relation to surrounding bony structures in normal subjects were investigated using magnetic resonance imaging. OBJECTIVES: This study determined the normal anatomic parameters of the lumbosacral nerve root and dorsal root ganglion, to which degenerative or pathologic changes may be compared. SUMMARY OF BACKGROUND DATA: In the previous literature, most authors have used various modalities in either cadavers or symptomatic patients to study the anatomic details of the lumbar nerve roots and dorsal root ganglia. The data in the literature are conflicting, mainly because of individual variations and different degrees of degenerative change in the spine. METHODS: Twenty male volunteers who had no back pain or radiculopathy underwent magnetic resonance imaging. Ages ranged from 22 to 38 years, with a mean of 30.4 years. T1-weighted coronal magnetic resonance images were taken from L1 to S1. Two hundred thirty-three nerve roots were examined, including 36 L1, 40 L2, 40 L3, 39 L4, 40 L5, and 38 S1. nerve roots Measurements were determined using a computer digitizer. RESULTS: The nerve root origin was at a more cephalad level for the caudad nerve roots, particularly the S1. The take off angels acutely changed at L1 and S1. The length of the nerve roots increased progressively to a maximum at L5, and decreased at S1. The center of the dorsal root ganglion was positioned more cephalad at S1. The average dimension of the dorsal root ganglion gradually increased from L1 to S1. The most striking difference was in the S1 root, which takes off more cephalad, at a more vertical angle, and has the shortest length of any of the nerve roots. The S1 dorsal root ganglion was also unique in that it was the largest and more frequently located intraspinally. CONCLUSION: The anatomy of the lumbar nerve roots and dorsal root ganglia and their relations to bony structures have been better defined in this study. Because of its more medial location, S1 radiculopathy may involve both the nerve root and dorsal root ganglion as a result of either disc herniation or degenerative L5-S1 facet changes. The relatively larger dorsal root ganglia and the greater dorsal root ganglion/foramen height ratios in the lower lumbar region may explain the higher incidence of L5 or S1 radiculopathy, particularly given the propensity to disc degeneration and intervertebral foraminal narrowing in the lower lumbar region. PMID- 8724083 TI - Morphometric study of myelinated fibers in human cervical spinal cord white matter. AB - STUDY DESIGN: Using human autopsy spinal cord specimens, morphologic measurements of myelinated nerve fibers were performed, focusing on the regions that include the main white matter conduction paths. The hemilateral spinal cord morphology was also measured, and its relation with the component myelinated nerve fibers determined. OBJECTIVES: To determine the relation between spinal cord transverse area in the normal lower cervical spine, the site most vulnerable to chronic compressive myelopathy, and myelinated nerve fibers. SUMMARY OF BACKGROUND DATA: Considerable interindividual variation normally is observed in the morphology of the spinal cord transverse area. The influence of this variation on the composition of the white matter myelinated nerve fibers is obscure. METHODS: The C7 segments from seven cadavers were resected, and from magnified photographs of paraffin-embedded specimens, the hemilateral spinal cord area and funicular area were measured. Nerve fiber morphology was measured using Epon-embedded specimens. Three regions that included the main conduction paths were sampled, and magnified photographs obtained. The nerve fiber transverse morphology was measured using the ellipse conversion method, and the myelinated nerve density and fiber area were determined. RESULTS: Marked interindividual variations were found in both the hemilateral spinal cord transverse area and funicular area. A positive correlation was noted between the two, with the spinal cord transverse area large in the cases with a large funicular area. For fiber density and area, histograms were constructed that showed characteristic distribution patterns in each region. By dividing each region into two components (i.e., small- and large-diameter fibers), it was found that the interindividual variation in large-diameter fiber density was small, clarifying that the absolute number of large-diameter fibers compared to fiber density is more strongly dependent on the funicular area. CONCLUSIONS: The absolute number of large-diameter myelinated fibers is smaller in cross-sections of thin as compared to those of thick spinal cord. When elucidating the pathophysiology of compressive myelopathy, it is necessary to study not only the circumstances surrounding the spinal cord, but this kind of factor intrinsic to the spinal cord itself. PMID- 8724084 TI - Anatomic considerations for posterior iliac bone harvesting. AB - STUDY DESIGN: This study analyzed bony features of the posterior ilium and relevant vital structures using cadavers and dry ilium specimens. OBJECTIVES: To determine quantitatively the safely zone of the posterior ilium and relevant vital structures with regard to bone graft harvesting. SUMMARY OF BACKGROUND DATA: The most frequently used site for bone graft harvesting is the posterior ilium. However, complications related to posterior iliac bone harvesting, such as donor site pain, neurovascular injury, instability of the sacroiliac joint, and herniation of abdominal contents, are still major concerns. Very little research with regard to the quantitative study of the posterior ilium has been reported. METHODS: Six cadavers (four male, two female) were used for the first part of this study. The posterior superior iliac spine was determined as a reference landmark. The distances from the posterior superior iliac spine to the superior cluneal nerves, the gluteal line, and the superior gluteal vessels were measured. The second part of the study involved 30 adult, dry iliac bony specimens. The posterior iliac region (extra-articular portion) was divided into three zones, and the corresponding dimensions of these zones were measured. RESULTS: The average distances from the posterior superior iliac spine to the superior cluneal nerves, gluteal line, and superior gluteal vessels were 68.8, 26.6, and 62.4 mm, respectively. The average width, height, and maximum thickness for Zone 1 were 34, 27.8, and 17.1 mm, respectively; the measurements for Zone 2 were 16.5, 31.8, and 14.2 mm, respectively. The average height for Zone 3 was 20.4 mm, and the average maximum thickness was 16.8mm. CONCLUSION: The ideal area of the posterior ilium for bone graft harvesting was found in Zone 1. Zones 2 or 3 may be considered it a greater quantity of cancellous bone graft is required; however, the risk of injury to the sacroiliac joint and superior gluteal vessels in these zones is increased. PMID- 8724085 TI - Quantified pain drawing in subacute low back pain. Validation in a nonselected outpatient industrial sample. AB - STUDY DESIGN: The criterion and construct validities of pain drawing, quantified by a simple total body area score of pain extent (area raw extent assessment score), were analyzed prospectively on consecutive patients (n = 103), drawn from a predefined blue collar worker population, all sick listed for 6 weeks as a result of low back pain. OBJECTIVES: To evaluate the validity of pain drawing as a screening tool in the secondary prevention of subacute low back pain. SUMMARY OF BACKGROUND DATA: Pain drawings have been used clinically for more than 40 years as a complement to a patient's verbal pain descriptions. The main objectives have been to differentiate functional pain from organic pain and to identify meaningful features in spatial-anatomic pain distribution. The ability of the pain drawing to delineate concurrent psychopathology correctly has been questioned. There is no consensus on which scoring method should be used. METHODS: The area raw extent assessment score was analyzed concurrently against the penalty point system and predictively against return to work and absenteeism over a period of 2 years. Content and construct validity assessed the relative influence of medical, psychologic, and subjective disability as well as psychosocial factors. RESULTS: Criterion validation of the area raw extent assessment score showed significant correlations, both concurrently against the penalty point score (r = 0.86, P < 0.001, with explained variance R2 = 0.75, P < 0.001) and predictively against occupational handicap (r = 0.48, P < 0.001). In construct validation, the highest explained variance was shown for medical (R2 = 0.46, P < 0.001) and psychologic factors (R2 = 0.46, P < 0.001) and psychologic factors (R2 = 0.34, P < 0.001) and for subjective disability (R2 = 0.32, P < 0.001). Variance in the area raw extent assessment score also was explained by psychosocial factors (R2 = 0.19, P < 0.01). CONCLUSIONS: Pain drawing quantification of the extent of pain shows high criterion and construct validity for the area raw extent assessment score. Content validity could be shown for significant clinical aspects of the disability experience--assets preferred for a screening tool in secondary prevention. PMID- 8724086 TI - Biomechanical analysis of thoracolumbar interbody constructs. How important is the endplate? AB - STUDY DESIGN: A biomechanical study of human cadaveric thoracic vertebral bodies was conducted using several anterior fusion options subjected to axial loads. This study emphasized the contribution of the endplate to resistance of graft subsidence. OBJECTIVES: To determine the importance of the vertebral endplate in resisting subsidence of various constructs into the vertebral body; the relative efficacy of potential alternative graft constructs such as iliac crest, ribs, humerus, and titanium mesh cage; and the importance of bone mineral content, vertebral level, and cross-sectional graft area on construct subsidence. SUMMARY OF BACKGROUND DATA: As the fixation length of anterior and posterior spinal constructs is reduced, load sharing of the anterior column has become more important to reduce failure of the shorter devices. Several alternative graft constructs and surgical techniques have been used for reconstruction of the anterior column. There exist little comparative data as to whether any of these constructs are superior and whether the vertebral endplate contributes significantly to the integrity of the construct. METHODS: Sixty-three isolated human cadaveric vertebral bodies from T3 to T12 were used to test seven different constructs in direct axial load onto prepared endplates with an electrohydraulic testing device. These constructs were: 1) titanium mesh cage (17 x 22 mm) on intact endplate, 2) C-shaped humerus on intact endplate, 3) tricorticated iliac graft in "tee configuration" on intact endplate, 4) tricorticated iliac graft in cancellous trough, 5) triple rib strut graft, 6) single rib on endplate, and 7) single rib on cancellous body. Dual X-ray absorptiometry assessment of bone mineral content was performed. A uniaxial load was applied with force and displacement data collected to determine maximal load to "failure" of the vertebral body. RESULTS: Preservation of vertebral endplate did not significantly increase the resistance to graft subsidence. The titanium cage construct provided the greatest resistance to axial load. CONCLUSIONS: Preservation of the vertebral endplate may not offer a significant biomechanical advantage in reconstructing the anterior column. Several alternative constructs are mechanically equivalent. PMID- 8724087 TI - Effect of specimen fixation method on pullout tests of pedicle screws. AB - STUDY DESIGN: Experimental axial pullout tests of a new type of pedicle screw were done on cadaveric lumbar vertebrae. The manner in which specimens were secured in the testing apparatus was varied to determined influence of specimen fixation method on the maximum pedicle screw pullout force. OBJECTIVES: To determine the appropriateness of embedding (i.e., potting) spinal specimens in polymer resin (e.g., bone cement or Plastic Padding [Plastic Padding Ltd., High Wycombe, Buckinghamshire, England]) for axial pullout tests of pedicle screws. Several different specimen fixation methods were examined to make recommendations for the standardization of future experimental testing protocols. SUMMARY OF BACKGROUND DATA: Axial pullout of transpedicular screws, although not a likely clinical mode of failure, is a popular experimental testing mode for evaluating screw-bone biomechanics. A wide variety of techniques for securing a vertebral specimen to counter the axial pullout force has been reported (including the use of polymer resin) with a correspondingly wide range in the resulting axial pullout strengths. The possible influence of the specimen fixation method on pedicle screw axial pullout strength has not been addressed previously. METHODS: Axial pullout tests of pedicle screws (DDS, Plus Endoprothetik, Rotkreuz, Switzerland) from the pedicles of 21 isolated lumber vertebral bodies were done using a Model 810 MTS Universal Testing Machine (MTS Systems, Inc., Minneapolis, Minnesota). The specimens were secured in a custom-made vise fixture either as is or after the vertebral bodies were potted in Plastic Padding up to the pedicle origin. Some of the potted specimens were wrapped first in latex to prevent polymer resin intrusion, and the others were unprotected. Pullout tests were attempted on both the left and right pedicles of each specimen, and the maximum pedicle screw pullout force was recorded. Measurement of bone mineral density by means of dual energy x-ray absorptiometry, in addition to macroscopic and scanning electron microscopy histologic analyses, microradiography, and energy dispersive X-ray spectroscopy, was done post-test to assist in the interpretation of the data. RESULTS: The maximum pedicle screw pullout force was found to be dependent on both the bone mineral density and the mode of fixation of the vertebrae. Embedding in polymer resin without protection of the specimen (i.e., latex wrapping) led to several instances of well-documented polymer resin intrusion; in these specimens, mean maximum pedicle screw pullout force was significantly greater than that of specimens secured without polymer resin and that of embedded specimens for which intrusion did not occur. CONCLUSIONS: Polymer resin intrusion can have a significant effect on the biomechanical characteristics of the bone-pedicle screw interface. When polymer resins are used to secure vertebral specimens for in vitro biomechanical tests of the bone pedicle screw interface, it is important to either prevent intrusion (e.g., with a latex wrapping) or document post-test (e.g., through the methods described in this article) that intrusion did not occur for the specimens included in the analysis. PMID- 8724088 TI - Increased limb lengths in patients with shortened spines due to tuberculosis in early childhood. AB - STUDY DESIGN: The spine and limb lengths of 26 patients who had a severe spinal deformity due to tuberculosis in childhood were measured and compared with similar data from 79 normal adult volunteers. OBJECTIVES: The aim of the study was to assess whether there are any compensatory growth mechanisms present in patients who had stunted spinal growth in childhood. SUMMARY OF BACKGROUND DATA: Previous studies have documented increased leg lengths in patients with adolescent idiopathic scoliosis who have had spinal fusions done. The aim of this study was to ascertain if this response was unique to scoliosis or was a more general response to the interruption of normal spine growth. METHODS: The standing height, spine height, leg length, and upper limb length were measured in 26 adults with stunted spinal growth due to tuberculosis of the spine contracted in early childhood, and compared with similar measurements in 79 normal volunteers. RESULTS: The patients with spinal deformity due to tuberculosis had significantly shorter mean standing and spine heights compared to the volunteers. However, the mean leg length of these patients was 19.4 mm longer than the volunteers and their mean upper limb length was 18.9 mm longer than the volunteers. These differences were statistically significant. CONCLUSION: Patients whose spinal growth was stunted due to disease in childhood have longer legs and upper limbs than healthy people. A compensatory stimulatory growth mechanism may be responsible for this. This has implications for the whole gamut of childhood spinal disorders that result in stunted spinal growth. PMID- 8724089 TI - Cost-effectiveness of lumbar discectomy for the treatment of herniated intervertebral disc. AB - STUDY DESIGN: A cost-effectiveness analysis of lumbar discectomy based on existing efficacy data and newly gathered cost data. OBJECTIVES: For patients with herniated lumbar discs unresponsive to conservative management, surgery relieves pain more rapidly but at higher costs than continued medical therapy. We evaluated the cost-effectiveness of lumbar discectomy for these patients. SUMMARY OF BACKGROUND DATA: Effectiveness estimates were based on the results of a published trial of 126 herniated disc patients randomized to surgical or nonsurgical treatment. Quality of life values were based on a study of 83 subjects with low back pain. Treatment costs for herniated discs were estimated from insurance data for 372 patients treated surgically and 1,803 treated medically. METHODS: Efficacy results were weighted by quality of life values to estimate the quality-adjusted benefit of surgery. Cost-effectiveness was calculated in dollars per quality-adjusted year of life gained. Supplemental data sources for cost and effectiveness provided ranges for sensitivity analyses. RESULTS: Surgery increased average quality-adjusted life expectancy by 0.43 years during the decade following treatment, a benefit similar to extending a healthy life by 5 months. Reimbursements for surgical patients were $12,550 more than for medical patients. Nondiscounted and 5% discounted cost-effectiveness were $29,200 and $33,900 per quality-adjusted year of life gained. Supplemental analyses confirmed the basecase effectiveness estimates but suggested that the cost of discectomy was overestimated. Replacing the main cost estimate with one based on HMO patients lowered discectomy's cost to $12,000 per quality-adjusted life-year gained. CONCLUSION: For carefully selected patients with herniated discs, surgical discectomy is a cost-effective treatment. Discectomy's favorable cost effectiveness results from its substantial effect on quality of life and moderate costs. PMID- 8724090 TI - Predictors of bad and good outcome of lumbar spine surgery. A prospective clinical study with 2 years' follow up. AB - STUDY DESIGN: Based on prospective assessment, patients with lumbar disc surgery were examined to determine reliable predictors for clinical outcome. OBJECTIVES: The prognostic value of a screening checklist developed in a previous study was evaluated in a 2-year follow-up. SUMMARY OF BACKGROUND DATA: Outcome studies of lumbar disc surgery document a success rate between 49-90%. It has been shown that a number of medical history data and sociodemographic and psychodiagnostic findings are of prognostic value for the outcome of lumbar spine surgery. METHODS: In addition to clinical and neuroradiologic examinations, 164 patients took part in a standardized interview. Eighty-two percent participated in a follow-up performed 2 years after the operation. Preoperative findings, outcome, and prediction of three diagnostic subgroups were compared. Eighty-three (51%) patients had disc herniation only, 29 (18%) had disc herniation and other relevant back diagnoses, and 51 (31%) had no disc herniation but had other relevant back diagnoses. RESULTS: In patients with disc herniation only, good results were observed in 53%, moderate in 19%, and bad in 28%. The accuracy of prediction of the postoperative result was 75% for the patients with good outcome and 86% for those with bad outcome. In the group of patients with diagnoses other than disc herniation, the success rate of the operation was 38% good, 28% moderate, and 41% bad, but the predictor score was not as useful as for the other groups. CONCLUSION: Patients with a high risk of a bad operation outcome after lumbar discectomy could be identified preoperatively. It is suggested that those patients take part in a pain management approach instead of or in addition to surgical intervention. PMID- 8724091 TI - Factors predicting employment 1 year after traumatic spine fracture. AB - STUDY DESIGN: This prospective cohort study evaluates the employment status of 489 persons after traumatic spine fracture. OBJECTIVES: To determine the rate, type, and predictors of employment 1 year after traumatic spine fracture. SUMMARY OF BACKGROUND DATA: The limited existing literature regarding employment after spine fracture reports variable return-to-work rates, tends to be retrospective, and generally evaluates a limited number of predictor factors at a time. METHODS: Four hundred eighty-nine persons ranging in age from 15 to 64 years who had experienced a spine fracture were assessed by a single examiner at hospital discharge and 1 year postinjury. Employment status and type. discharge neural and functional status, pain level, demographics, injury level and severity, and early treatment details were evaluated. RESULTS: At 1 year postinjury, 54% of subjects were working. A higher percentage of the employed were working part time and for fewer weeks per year than preinjury. A higher percentage were working at unskilled clerical, sales, or service jobs than preinjury. The significant positive (+) and negative (-) predictors of employment were (from strongest to weakest); worked in year previous to injury (+); employed at time of injury (+); Worker's compensation Board coverage (-); spinal fracture surgery (+); high-level spine fracture (-); pain (-); Functional Independence Measure score (+); and days of stay in intensive care unit and spinal unit (-). CONCLUSIONS: For the first year after spinal fracture, unemployment is common. Those who do return to work are more likely to modify the amount and type of work they do and to have been employed preinjury. PMID- 8724092 TI - First-time operation for lumbar disc herniation with or without free fat transplantation. Prospective triple-blind randomized study with reference to clinical factors and enhanced computed tomographic scan 1 year after operation. AB - STUDY DESIGN: This prospective triple-blind randomized study of 99 patients concerned the use of free fat transplantation for operation for lumbar disc herniation. OBJECTIVE: To subsequently examine the patients after median 376 days who were subjected to enhanced computed tomographic scan. SUMMARY OF BACKGROUND DATA: In studies on experiments with animals, the degree of intraspinal scar tissue has shown to be reduced in connection with free fat transplantation. Scar tissue is seen frequently after operation for lumbar disc herniation, but it is uncertain whether the scar tissue can lead to symptoms. METHODS: The clinical outcome was scored using the Low Back Pain Rating Scale. Enhanced computed tomographic scan was assessed regarding the degree of scar tissue and survival of fat transplant. RESULTS: There was no different in the clinical outcome between the two groups. Significantly fewer patients had dural scarring in the group who had a fat transplantation, but there was no difference regarding the degree of radicular scarring. The transplant was shown on computed tomographic scan at the follow-up examination in 66% of the patients who had a fat transplantation. CONCLUSIONS: Free fat transplantation can reduced the degree of dural scar tissue after operation for lumbar disc herniation but does not result in a clinically better outcome. PMID- 8724093 TI - Nonorganic signs of significance in low back pain. AB - STUDY DESIGN: Medical records and imaging studies including one or more interviews and physical examinations by the author were reviewed in 878 people remaining out of work for 13 weeks or more because of occupational low back pain. OBJECTIVES: To assess the prognostic value of nonorganic signs, including those validated by Waddell et al and two others previously undescribed, limitation of shoulder motion with production of low back pain, and low back pain resulting from movement limited to the cervical spines. SUMMARY OF BACKGROUND DATA: Review of all records of people evaluated by the author over a 3-year period (1990-1993) provided the material for this study. All available medical, insurance, and legal records were reviewed, including reports or examination results of radiographic films, or both. In the majority, more sophisticated investigation had been performed with complete reports of the studies themselves being available to the author. METHODS: A complete history and physical examination were accomplished by the author on each individual, including evaluation of each of the test results for nonorganicity. Medical records, including radiographic films, magnetic resonance imaging, computerized axial tomography, myelography, bone scan, electromyography, etc., were collated. Statistical analysis of results followed. RESULTS: The finding of limitation of shoulder motion resulting in low back pain indicated a worse prognosis for return to work than that of positive Waddell signs alone, 69.6% versus 52.9%. If cervical motion additionally produced low back pain, the outlook was poorer (no return to work in 73.1%). CONCLUSION: When present with a triad of the signs validated by Waddell, the new signs significantly increase the predictability of failure to return to work by workers with compensable low back pain. PMID- 8724094 TI - Symptomatic spinal hemangiomas in association with cutaneous hemangiomas. A case report. AB - STUDY DESIGN: This case report describes a patient with associated vertebral and peripheral vascular malformations presenting with a painful scoliosis. OBJECTIVES: To treat the vertebral osseous lesion successfully with embolization. SUMMARY OF BACKGROUND DATA: Although vertebral vascular malformations are well described, the association with peripheral lesions is uncommon. When associated peripheral vascular malformations occur, they are usually in tissues derived from the same embryonic segment, which was not the case in this instance. METHODS: After investigation with angiography, this patient's vertebral osseous lesion was treated by embolization. RESULTS: This patient's pain has been abolished since embolization. Her scoliosis has not progressed during follow-up. CONCLUSIONS: This case illustrates the association between spinal and peripheral vascular malformations, and that this association is not always metameric. It highlights the fact that invasive procedures other than surgery are a valuable part of the therapeutic armamentarium. PMID- 8724095 TI - Tortuosity of the vertebral artery in patients with cervical spondylotic myelopathy. Risk factor for the vertebral artery injury during anterior cervical decompression. AB - STUDY DESIGN: The case report presented herein shows tortuosity of the vertebral artery in a patient with cervical myelopathy. This case led the authors to evaluate 22 other patients who also had undergone anterior cervical fusion. They were studied before operation by either magnetic resonance imaging angiography or selective vertebral angiography. OBJECTIVES: To analyze the radiographs, computed tomography, magnetic resonance imaging, and angiography findings to detect any tortuosity of the vertebral artery in patients with cervical myelopathy to show the risk factors of vertebral artery injury during anterior decompression. SUMMARY OF BACKGROUND DATA: Complications of vertebral artery laceration during cervical anterior decompression are rare, so this injury and abnormality in the course of vertebral artery in patients with cervical myelopathy receive little attention. METHODS: The tortuosity of the vertebral artery was assessed by angiography, magnetic resonance imaging, and computed tomography. RESULTS: Mild vertebral artery tortuosity was observed in 10 patients and loop formation in three associated with cervical spondylotic changes. CONCLUSIONS: This study suggests that vertebral artery loop formation is developed associated with cervical spondylotic changes. During the anterior decompression of cervical spondylotic myelopathy or radiculopathy, the looped vertebral artery could be injured by an excessive wide rejection of the bone or disc material. In the case of vertebral artery migration, the looped vertebral artery can even be injured by routine procedures. PMID- 8724096 TI - Retroperitoneal pseudomeningocele complicated by meningitis following a lumbar burst fracture. A case report. AB - STUDY DESIGN: This case report demonstrates an unusual complication after anterior decompression and fusion of a lumbar burst fracture. OBJECTIVES: The treatment of this patient involved placement of a computed tomography-guided percutaneous drain and intravenous antibiotics to treat an infected retroperitoneal pseudomeningocele. SUMMARY OF BACKGROUND DATA: A case of an anterior retroperitoneal pseudomeningocele complicated by meningitis is presented. This pseudomeningocele occurred in a patient after an L3 burst fracture associated with a dural laceration. METHODS: The patient was admitted to the authors' trauma unit after a motor vehicle accident with an acute L3 fracture associated with incomplete paraplegia. He underwent an urgent anterior corpectomy, strut grafting, and instrumentation. At surgery, he was noted to have a large anterior dural laceration. After surgery, a large retroperitoneal pseudomeningocele developed that became infected with Staphylococcus epidermidis. RESULTS: After placement of a computed tomography-guided percutaneous drain and intravenous antibiotics, the pseudomeningocele resolved. His anterior fusion healed uneventfully and his neurologic deficit improved dramatically. CONCLUSIONS: A case of an anterior retroperitoneal pseudomeningocele complicated by meningitis is presented. This pseudomeningocele occurred in a patient after an L3 burst fracture associated with a dural laceration. The patient was treated successfully with computed tomography-guided percutaneous drain placement and intravenous antibiotics. He made an excellent functional recovery after a severe neurologic injury. PMID- 8724097 TI - Multifocal metachronous epidural abscesses of the spine. A case report. AB - STUDY DESIGN: A retrospective case report of a patient who had a lumbar epidural abscess treated surgically who then developed a cervical epidural abscess that also required surgical treatment. OBJECTIVES: To describe a patient in whom treatment of a single epidural abscess with surgery and antibiotics was not sufficient to eradicate the systemic infection. SUMMARY OF BACKGROUND DATA: Epidural abscesses are most commonly seen after invasive procedures that violate the epidural space. Epidural abscesses are usually a solitary event occurring in only one location and are usually treatable with surgical drainage and parenteral antibiotics. METHODS: An elderly patient presented with neck and shoulder pain and fever. Evaluation revealed degenerative disease of the cervical spine. Within a week, she developed a cauda equina syndrome secondary to a lumber epidural abscess. The abscess was drained and intravenous antibiotics were given. Seventeen days later, while still receiving antibiotics, she developed a cervical epidural abscess which also required surgical drainage. RESULTS: The patient showed gradual improvement in her neurologic status. No recurrence of either epidural abscess was observed. CONCLUSIONS: An epidural abscess may represent a serious systemic infection that requires aggressive treatment. Close follow-up is necessary to ensure that the infection has been eradicated and that no recurrent abscess has formed in the same or a different location. Aggressive antibiotic treatment is also strongly recommended. PMID- 8724098 TI - Spinal stenosis above a healed tuberculous kyphosis. A case report. AB - STUDY DESIGN: This case report illustrates two patients with paraplegia of late onset resulting from cord compression in the hyperlordotic thoracic or thoracolumbar spine proximal to a healed tuberculous kyphosis. OBJECTIVES: The objective of this paper is to highlight that degenerative stenosis of the thoracic spine proximal to a healed stable kyphosis can be a cause of paraplegia of healed disease. SUMMARY OF BACKGROUND DATA: Paraplegia resulting from tuberculosis of the spine can be of early or late onset. Early onset paraplegia is usually a result of cord compression by active disease. Late onset paraplegia can be due to disease reactivation, bony ridge compression, or unstable kyphosis. To our knowledge, compression of the cord proximal to a healed, stable kyphosis giving rise to paraplegia has never been reported. METHODS: Patient 1 presented with symptoms of spinal claudication and progressive paraparesis. He was found to have spinal stenosis in the hyperlordotic thoracolumbar spine proximal to a healed lumbosacral tuberculous kyphosis. Patient 2 presented with a 2-year history of progressive paraplegia. Imaging revealed cord compression at the hyperlordotic T10-11 segment by disc protrusion, and facet hypertrophy. RESULTS: Laminectomy successfully relieved the first patient of all symptoms but the second patient had significant deterioration of the neurologic status after surgery. CONCLUSION: Compensatory hyperlordosis of the thoracic or thoracolumbar spine commonly occurs in patients with severe tuberculous thoracolumbar or lumbosacral kyphosis. Degenerative spinal stenosis and cord compression at such hyperlordotic segment can cause late onset paraplegia. The blood supply of these chronically compressed cords is precarious and the risk of surgery is high. PMID- 8724099 TI - Chemonucleolysis. AB - Even with a history of controversy and troubling complications, chymopapain has endured the test of time to show 30 years of clinical success in the treatment of herniated nucleus pulposus. Strict attention to indications, contraindications, and technique ensures safety and efficacy of treatment. A trend to decreased dosage may result in less postinjection spasm. Between 1982-1991, 121 adverse events in 135,000 patients were reported to the Food and Drug Administration and investigated. Seven cases of fatal anaphylaxis, 24 infections, 32 bleeding problems, 32 neurologic events, and 15 miscellaneous occurrences were found. Overall mortality rate was 0.019%. All categories were of lesser incidence than complications with laminectomy. Long-term results show that improvement after chemonucleolysis is maintained, whereas the outcome after laminectomy is reported to deteriorate with time. Cost savings with chemonucleolysis over laminectomy are largely a matter of shorter hospitalization. A protocol for cervical chemonucleolysis is being developed in the United States after good results have been shown in Spain and France. Other enzymes continue under investigation, but chymopapain remains the standard to which they are compared. PMID- 8724100 TI - Sir William Gowers 1845-1915. AB - This is a brief review of the life and contributions to neurologic science of Sir William Gowers. He was a neurologist in England in the late 19th century who, before the availability of modern diagnostic aids, established the importance of clinical examination, including history and physical signs, in the diagnosis of neurologic disorders. He was the first neurologist to identify a removable spinal tumor. He arranged for his colleague, Sir Victor Horsley, to remove it and the operation was completely successful. He invented the patella hammer and his book "Manual of Diseases of the Nervous System" (in two volumes) was the standard reference until early in the 20th century. PMID- 8724101 TI - Controversy in spine care. Is fusion necessary after anterior cervical discectomy? AB - Anterior cervical discectomy is an effective and reliable treatment for nerve root or cord compression caused by disc herniation or spondylosis. Although physicians have traditionally included fusion as a part of this procedure, recent experience has suggested that this may not be necessary. Dr. Volker Sonntag and Dr. Peter Klara express opposing views on the need for fusion after discectomy and support their perspectives with clinical experience and a review of the pathoanatomy of disc disease. Dr. Sonntag believes that the majority of patients are well served with discectomy alone, avoiding the complications of graft harvest and potential nonunion. Dr. Klara feels that the interposed graft restores foraminal height and maintains cervical lordosis, both of which are important to a good outcome. PMID- 8724102 TI - Assessment of spinal fusion. Critical evaluation of imaging techniques. AB - Before any radiologic imaging modality is employed in an investigation study, its efficacy must be critically assessed. The purpose of this report is to demonstrate how the choice of computed tomography imaging parameters affects the information provided by a computed tomographic examination. It is apparent from these results that imaging parameters must be optimized before the results of an imaging technique can be compared to other modes of diagnostic evaluation, including surgical observations. PMID- 8724103 TI - Effects of spinal flexion and extension exercises and their associated postures in patients with acute low back pain. PMID- 8724104 TI - Effects of spinal flexion and extension exercises and their associated postures in patients with acute low back pain. PMID- 8724105 TI - Effects of spinal flexion and extension exercises and their associated postures in patients with acute low back pain. PMID- 8724106 TI - The effects of the apolipoprotein B signal peptide (ins/del) and XbaI polymorphisms on plasma lipid responses to dietary change. AB - There is interindividual variation in plasma lipid response to dietary changes. The polymorphisms which are associated with plasma lipid levels could possibly explain part of this variation. Therefore, the apolipoprotein B (apo B) signal peptide insertion/deletion (ins/del) and XbaI restriction fragment length polymorphisms are possible regulators of plasma lipid responses. We examined their role in the regulation of plasma lipid responses in 87 North Karelians (43 men, 44 women). The dietary study consisted of a 2-week baseline period (34-35% of energy from fat), followed by an 8-week low-fat (24 En%), low-cholesterol (279 mg/d) diet period and an 8-week switchback period. In this study population the apo B ins/del and XbaI polymorphisms exhibited mainly similar and partly significant effects on the responses of plasma very low-density lipoprotein (VLDL) and high density lipoprotein2 (HDL2) cholesterol to dietary changes. After consumption of the low saturated fat, low-cholesterol diet, ins/ins X - /X - homozygotes showed the greatest increase in VLDL cholesterol (p < 0.05 for differences between ins/del genotypes) and the greatest fall in HDL2 cholesterol (p = 0.01 for ins/del and p = 0.05 for XbaI), while only minimal alterations were seen in the del/del and X + /X + groups. After returning to the original diet, the changes of these lipids were reversed, ins/ins and X -/X - homozygotes having the greatest reductions in VLDL cholesterol (p < 0.05 for XbaI) and the greatest increases in HLDL2 cholesterol (p < 0.001 for XbaI). The findings suggest that plasma VLDL and HDL2 cholesterol responsiveness to diet may be partly explained by variation at the apo B gene. PMID- 8724107 TI - Lipoprotein(a) and peripheral atherosclerosis in older adults. AB - As part of an ancillary study to the Systolic Hypertension in the Elderly Program, carotid and lower extremity arterial disease (LEAD) were evaluated in 369 subjects, 186 with a systolic blood pressure (SBP) > or = 160 mmHg, and 183 with SBP < 160 mmHg. Both groups had a diastolic blood pressure (DBP) < 90 mmHg. Internal carotid stenosis was identified by Doppler and LEAD was assessed using the ankle to arm systolic blood pressure ratio, commonly called the ankle/arm index (AAI). Lp(a) values were obtained from frozen sera and values > or = 20 mg/dl were considered elevated. Rates of carotid stenosis were 24% among those with an Lp(a) > or = 20 mg/dl and 14% among those with an Lp(a) level < 20 mg/dl (P = 0.020). The relationship between Lp(a) and LEAD was even stronger. Those with an Lp(a) > or = 20 mg/dl had a 36% prevalence of a low AAI vs 14% among those with a Lp(a) level < 20 mg/dl (P < 0.001). Lp(a) values were also associated with the severity of LEAD. Controlling for other risk factors did not reduce the association between either LEAD or carotid stenosis and an Lp(a) > or = 20 mg/dl. Thus, Lp(a) appears to be independently associated with peripheral atherosclerosis in older adults, both men and women. The relationship is particularly strong for atherosclerotic disease of the lower extremities. PMID- 8724108 TI - Association between the LPL-D9N mutation in the lipoprotein lipase gene and plasma lipid traits in myocardial infarction survivors from the ECTIM Study. AB - Using polymerase chain reaction (PCR) based techniques, we have identified individuals in the ECTIM study of myocardial infarction survivors (cases) and healthy matched controls who are carriers for a mutation of the gene for lipoprotein lipase (LPL) which alters amino acid 9 from aspartic acid to asparagine (LPL-D9N). The frequency of carriers in the cases from Belfast and France (3 separate centres) was 2.5 and 3.7%, respectively (mean 3.3%, 95% CI 1.9 4.7) and in the controls 2.0 and 2.9%, respectively (mean 2.7%, 95% CI 1.6-3.8%), but this difference was not statistically significant. In the cases, carriers of the allele for LPL-N9 had higher levels of several plasma lipid traits including total triglycerides (TG) (30%), very low density lipoprotein (VLDL) cholesterol (19%), apo E (24%), apo C-III (17%), lipoprotein particles (Lp) containing both apo E and apo B (LpE:B) (32%), and particles containing both apo C-III and apo B (LpCIII:B) (39%), and this effect was consistent in cases both from Belfast and from the French centres combined. By contrast, in the controls there were no differences in any lipid trait between carriers and non-carriers of the mutation that was consistent between the French centres and Belfast. There were no significant differences in the levels of any measured factor between cases and controls that could explain the different effect on plasma lipid traits associated with the mutation. However, compared to the non-carriers, in both cases and controls who carried the mutation, plasma TG concentrations were higher in those whose body mass index (BMI) was above the mean of the sample (26.0 kg/m2), with statistically significant interaction seen between BMI and genotype and levels of apo C-III, and lipoprotein particles containing both apo C-III and apo B (P < 0.02). The data suggest that carriers for the LPL-N9 mutation have a mild genetic predisposition to developing hyperlipidaemia and an atherogenic lipid profile, but that this requires the presence of other genetic or environmental factors for full expression, one of which appears to be increasing obesity. PMID- 8724109 TI - Lipoprotein(a) is an independent risk factor for multiple cerebral infarctions. AB - In an attempt to ascertain whether Lp(a) is a risk factor for multiple cerebral infarctions (MCI), we have studied 83 patients with proven MCI and 39 subjects without MCI by computed tomography (CT). Seventy-one patients with non-insulin dependent diabetes mellitus (NIDDM) were included: 52 with and 19 without MCI. Serum Lp(a) levels were significantly higher in patients with MCI than in subjects without MCI. There were no differences in serum Lp(a) levels between NIDDM and non-diabetic patients with MCI. The logistic regression analysis revealed that Lp(a) and hypertension were independent risk factors for the cerebral event. The current study demonstrated that Lp(a) and hypertension are significant risk factors for multiple cerebral infarctions. PMID- 8724110 TI - Incomplete dominance of type III hyperlipoproteinemia is associated with the rare apolipoprotein E2 (Arg136-->Ser) variant in multigenerational pedigree studies. AB - In the process of screening apolipoprotein (apo) E genotypes in a population of subjects with lipid abnormalities, we have identified five subjects (one homozygote and four heterozygotes) with an abnormal 109 base pairs band following apo E restriction isotyping of amplified DNA with the restriction endonuclease CfoI. The polymerase chain reaction (PCR) products were cloned and their sequencing revealed a C-->A substitution at the first nucleotide of codon 136. This mutation resulted in an amino acid substitution Arg to Ser, previously described as apo E2 Christchurch. Family studies were carried out for four of the probands. In these kindreds, stepwise multiple regression analyses indicated that 78% of the cholesterol variability in men was predicted by body mass index, age and the rare apo E2 (Arg136-->Ser) variant. In women, age and the apo E2 (Arg136- >Ser variant predicted 54.9% of the variability in cholesterol levels. Linkage analysis suggested that the presence of the apo E2 (Arg136-->Ser) variant was linked with the occurrence of cholesterol enriched triglyceride rich lipoproteins and with an incomplete dominance of type III hyperlipoproteinemia. Our data indicates that this mutation may be a relatively common cause of dyslipidemia in the Spanish population. PMID- 8724111 TI - Blood pressure and heart rate response during exercise in men and women in the USA and Russia lipid research clinics prevalence study. AB - Examination of blood pressure (BP) and heart rate (HR) measurements at rest and during exercise in samples of USA and Russian middle-aged men and women show significant differences between countries for both genders. Russian men had higher resting systolic blood pressure (SBP) than USA men but lower SBP at both stages of exercise. Russian women had significantly higher resting SBP than USA women at rest and also during exercise. Russian men and women had significantly lower HR at rest and during exercise than USA men and women. Differences between countries were also noted for mean body weight, height, lipid levels, percentages of smokers and several other variables, and it was postulated that differences in these factors might be responsible in part for differences in SBP and HR during exercise. After adjustment for these variables, differences in SBP during exercise between USA and Russian men remained, but differences between USA and Russian women disappeared. For both genders, differences in HR during exercise remained after adjustment. Mortality analyses in USA and Russian men indicated that stage 2 SBP response during exercise was generally not a significant mortality risk factor after adjustment for age, BMI, TC, smoking and resting SBP. In a similar model, stage 2 HR response during exercise was also a non significant risk factor in mortality. PMID- 8724112 TI - Expression of VCAM-1 in rabbit iliac arteries is associated with vasodilator dysfunction of regenerated endothelium following balloon injury. AB - Previous studies have demonstrated impaired endothelium-dependent vasodilation following balloon injury of the rabbit iliac artery, suggesting dysfunction of the regenerated endothelium. More recently, expression of vascular cell adhesion molecule-1 (VCAM-1) has been shown up to 4 weeks in a different injury model of the rabbit aorta, suggesting sustained inflammatory activation of endothelium following injury. The aim of the present study was to combine the examination of VCAM-1 expression, as a marker of cellular activation, and the assessment of endothelium-dependent relaxation to test the hypothesis that different forms of altered endothelial function are concurrently present in the chronic phase following experimental balloon angioplasty. New Zealand White rabbits fed either a standard (n = 7) or a 1% cholesterol (n = 8) diet, underwent balloon injury of the iliac artery 5 weeks following the initiation of the diet. Four weeks after balloon injury, control and balloon-injured arteries were harvested for in vitro studies of vascular reactivity, for morphometric analysis and for immunocytochemical staining with Rb 1/9 monoclonal antibody directed against VCAM 1 and with CD 31 monoclonal antibody for the identification of endothelial cells. The combination of balloon injury and hypercholesterolemia resulted in a marked impairment of endothelium-dependent relaxation to acetylcholine and in a pronounced intimal proliferation compared to control or either intervention alone. Control rings of rabbits fed a normal diet did not reveal positive staining for VCAM-1. Balloon-injured rings of the animals fed a normal diet showed focal areas of positive staining in the superficial cell layer overlying intimal lesions. In the group fed a high cholesterol diet, control rings and ballooned rings showed positive staining for VCAM-1 in cells overlying intimal lesions. In all groups the superficial cell layers were identified as endothelial cells by positive staining for CD 31. In conclusion, the present study shows that regenerated endothelium following mechanical arterial injury reveals expression of VCAM-1 together with impaired receptor-mediated vasodilator capacity. Thus, the expression of VCAM-1 and the impairment of endothelium-dependent relaxation may represent different features of endothelial dysfunction following balloon injury which may actively influence the proliferative lesion of restenosis after balloon angioplasty. PMID- 8724114 TI - Prevalence of hyperapobetalipoproteinemia and factors affecting the phenotype expression in children and young adults. The Cardiovascular Risk in Young Finns Study. AB - Hyperapobetalipoproteinemia (hyperapoB) is one of the most common phenotypes in patients with premature coronary heart disease. In this study the factors that affect the expression of the hyperapoB phenotype were evaluated in young individuals. A cohort of 1125 children and young adults aged 9-24 years was classified into three groups by sex: (1) normal serum apolipoprotein B (apoB), (2) high apoB (> or = 90th percentile) and normal low density lipoprotein cholesterol (LDL-C < 90th), (3) high apoB and high LDL-C (> or = 90th percentile). In females, alcohol use (11, 33, 0%, in groups 1-3, P < 0.05) and oral contraceptive use (35, 83, 47%, P < 0.01) were significantly different between the groups and the highest frequencies were seen in the hyperapoB group (group 2). In both sexes smoking tended to be more common in the hyperapoB group (29, 43, 18%, P < 0.14). The two hyperapoB definition criteria (high apoB and low LDL-C/apoB ratio) were studied with multiple linear regression analyses. Oral contraceptive use correlated positively with apoB values (coefficient beta = 0.101, R2 = 2.1%, P < 0.01) and negatively with LDL-C/apoB ratio (beta = -0.134, R2 = 3.3%, P < 0.001). Alcohol use (beta = -0.072, R2 = 2.9%, P < 0.001) and smoking (beta = -0.050, R2 = 1.0%, P < 0.05) correlated negatively with LDL C/apoB ratio. Prevalence of the hyperapoB phenotype was 4.4%. According to the results, the expression of the hyperapoB phenotype may be influenced by common lifestyle habits. This should be considered if high risk young individuals are identified through the expression of the hyperapoB phenotype. PMID- 8724113 TI - Molecular and biochemical approaches in the identification of heterozygotes for homocystinuria. AB - We compared biochemical and molecular methods for the identification of heterozygous carriers of mutations in the cystathionine beta-synthase (CBS) gene. Eleven relatives of seven unrelated patients with homocystinuria due to homozygous CBS deficiency and controls were studied with respect to total homocysteine concentrations before and after methionine loading. In addition, we determined CBS activity in cultured skin fibroblasts and tested for the presence of five known mutations by a PCR-based method in these seven patients, their relatives and controls. The results demonstrate that measurement of homocysteine after methionine loading and assay of CBS enzyme activity in cultured fibroblasts identify most but not all heterozygotes. There was significant correlation between homocysteine concentrations and CBS activities only after methionine loading (r = 0.12, 0.48, 0.48 and 0.50 at 0, 4, 6 and 8 h, respectively). Among the homozygous patients, molecular approaches identified five T833C and two G919A mutations out of 14 independent alleles, confirming the studies of others that these represent the two most prevalent mutations. In addition, we found that three of six heterozygotes with the T833 C allele had post-methionine loading homocysteine levels which overlapped with controls and of the other three, one (as well as an obligate heterozygote who did not carry any of the five mutant alleles tested) had CBS activity comparable to that of controls. These findings demonstrate that genotyping is useful as an adjunctive method for the diagnosis of the heterozygous carrier state of CBS deficiency. PMID- 8724115 TI - Testosterone-induced suppression of lipoprotein(a) in normal men; relation to basal lipoprotein(a) level. AB - The concentration of lipoprotein(a) [Lp(a)] in human plasma is largely genetically determined and is inversely correlated to the size of apolipoprotein(a) [apo(a)]. Additionally, Lp(a) values are relatively stable within individuals and are only marginally susceptible to therapeutic treatment. The aim of our study was to evaluate the effect of exogenous testosterone on plasma Lp(a) concentration. The study was carried out on 19 healthy men who were receiving weekly intramuscular injections of 200 mg testosterone enanthate. Lp(a) values were determined at multiple time-points by a double monoclonal antibody based enzyme immunoassay. This method is not sensitive to variation in Lp(a) size and the values are expressed in nmol/l. Apo(a) size isoforms were determined by agarose gel electrophoresis followed by immunoblotting. No correlation was found between the baseline Lp(a) values and the baseline values of testosterone or estradiol. The Lp(a) response to testosterone treatment varied widely among subjects and was dependent upon the pretreatment Lp(a) concentration. For 10 subjects with low Lp(a) values (< 25 nmol/l), no significant decrease in Lp(a) was observed while, for the nine individuals with Lp(a) values > 25 nmol/l, there was a significant and consistent reduction in Lp(a) ranging from 25 to 59%. Lp(a) levels returned to baseline values following cessation of testosterone administration. Apo(a) size polymorphism did not appear to play a role in the determination of Lp(a) response to testosterone. PMID- 8724116 TI - Exercise-induced increase in lipoprotein (a). AB - In the Oslo Diet and Exercise Study (ODES) 219 healthy middle-aged physically inactive persons with moderately deranged risk factor levels (increased bodyweight, diastolic blood pressure, serum cholesterol, triglycerides, decreased HDL-cholesterol) were randomized to 4 intervention groups: dietary intervention, exercise, diet + exercise and control. The purpose of the study was to test if these interventions maintained for a year, isolated or in combination, would change coronary risk factor levels as compared to control. One of the risk factors included was lipoprotein (a) (Lp(a)). The hypothesis to be tested was if physical exercise would be associated with increased levels of Lp(a) as a result of intervention. Those who exercised increased their Lp(a) levels with 15.4 (S.E. = 8.0) mg/l as compared to no exercise (P < 0.05). Also, dietary intervention tended to increase Lp(a), but the increase did not reach statistical significance. There was no detectable interaction on the effect on Lp(a) of the two intervention modalities. A dose-response relationship was found between change in the exercise-specific variables heart rate and peak oxygen uptake, and Lp(a)-change and this dose-response was most pronounced in the exercise group. Change in Lp(a) was associated to change in several lifestyle related variables such as alcohol intake and waist circumference, pointing to the possibility that Lp(a), at least in some subpopulations, is more amenable to change through lifestyle alterations than reported so far. PMID- 8724117 TI - Triglyceride enriched lipoprotein particles correlate with the severity of coronary artery disease. AB - A group of 100 male normotensive, non-obese, non-diabetic subjects who had undergone coronary angiography were studied to determine relationship between the severity of coronary artery disease (CAD) and plasma lipids, apolipoproteins and lipoprotein particles defined by their apolipoprotein composition. CAD was found in 84 and no measurable lesions were found in 26 subjects. The severity of CAD was determined on the basis of size and number of lesions and expressed in terms of a global CAD score. Low density lipoprotein (LDL)-cholesterol showed a tendency to be higher in CAD patients than in CAD-free subjects (216 vs. 205 mg/dl, P = 0.07). HDL-cholesterol showed a tendency towards lower values in CAD patients compared to CAD-free subjects 35 vs. 41 mg/dl, P = 0.07). In univariate analysis the severity of CAD correlated with (i) complex, apolipoprotein (apo) B containing particles (Lp-B-complex, r = 0.31, P = 0.005), (ii) HDL-cholesterol (r = -0.30, P = 0.005), (iii) apoC-III in heparin precipitate (r = 0.30, P = 0.005) and (iv) plasma triglycerides (r = 0.25, P = 0.02), all of which are related to triglyceride-rich lipoproteins. A comparison between the two subspecies of complex lipoprotein particles revealed that those containing apolipoproteins B, C III and E (Lp-B:C:E complex) were more closely associated with CAD score (r = 0.27, P = 0.01) than those containing apolipoproteins A-II, B, C, D and E (Lp-A II:B-complex). LDL-cholesterol also correlated with the global CAD score (r = 0.23, P = 0.03). In multiple regression analysis only HDL-cholesterol (P = 0.003), apoC-III-ratio (P = 0.007), Lp-B-complex (P = 0.02) and Lp-B:C:E-complex (P = 0.04) showed significant correlation with CAD score. The results of this study demonstrate that some of the triglyceride rich lipoprotein particles represent a risk factor for CAD and support the clinical usefulness of specific assays capable of distinguishing lipoprotein particles on the basis of apolipoprotein composition. PMID- 8724118 TI - Genetic correlations between lipoprotein phenotypes and indicators of sex hormone levels in Mexican Americans. AB - Previous studies have shown that the inverse relationship between HDL cholesterol (HDL-C) and triglyceride (TG) levels, risk factors for cardiovascular disease, is due largely to the effects of shared genes. HDL-C and TG are also known to be related to endogenous sex hormone levels, however the nature of the relationships is unclear. The objective of this study is to ascertain the extent to which these relationships are determined by shared genes. We conducted a multivariate quantitative genetic analysis of HDL-C, TG, dehydroepiandrosterone sulfate (DHEAS) and sex hormone-binding globulin (SHBG) in 635 people from 27 pedigrees participating in the San Antonio Family Heart Study. Heritabilities (h2) and genetic and environmental correlations (rho G and rho E) were estimated simultaneously by maximum likelihood methods. All four traits showed significant (P < 0.05) heritabilities: h2HDL-C = 0.38, h2TG = 0.54, h2DHEAS = 0.43, h2SHBG = 0.26. Significant genetic correlations were detected between HDL and each of the other traits: rho G(HDL-TG) = -0.56, rho G(HDL-DHEAS) = 0.23 and rho G(HDL-SHBG) = -0.56. However, there were no significant genetic correlations between TG and either measure of sex hormones. Thus, at least three separate groups of genes influence HDL-C levels in Mexican Americans: one group that has pleiotropic effects on HDL and TG, one group influences both HDL-C and SHBG and a third influences both HDL-C and DHEAS. PMID- 8724119 TI - Lipoprotein(a) is increased in triglyceride-rich lipoproteins in men with coronary heart disease, but does not change acutely following oral fat ingestion. AB - Association of apo(a)/Lp(a) with triglyceride-rich lipoproteins (TGR-Lps) is determined by different factors that are poorly understood. Some previous studies suggested that apo(a) in TGR-Lps may affect the atherogenicity of the TGR particles. To study whether there are any peculiarities in postprandial (pp) Lp(a) metabolism, we have determined apo(a) phenotypes and Lp(a) concentrations in 46 subjects with coronary heart disease (CHD) and in six normolipidemic individuals at different time points (4, 6 and 8 h) following an oral fat tolerance test. While mean triglyceride concentration reached its maximum 6 h after a standardized fat meal, no change in total cholesterol and in mean Lp(a) plasma concentration was detected at any time point after the fat load. In 6 normolipidemic probands and in 8 patients with CHD, who were matched for apo(a) phenotype, lipoprotein levels, age and body weight, we followed the distribution of apo(a) in plasma density gradient fractions in the fasting and pp state. In the CHD patients a significant larger percentage of apo(a) reactivity was detected in TGR-Lps in the pre- as well as in the postprandial state, compared to control subjects. The fat intake did not induce a significant change of apo(a) reactivity in the TGR-Lp fractions in both groups. The apo(a) isoform-size and the Lp(a) plasma concentration in the fasting state had no influence on the individual variation of the Lp(a) concentration in pp TGR-Lp fractions. Our results provide evidence that TGR-Lp fractions of CHD patients are enriched in apo(a) reactivity compared to healthy controls, but do not support the hypothesis that Lp(a) acts atherogenically through a pp increase of its plasma concentration. PMID- 8724120 TI - Hypocholesterolemic effect of sesame lignan in humans. PMID- 8724121 TI - Distribution pattern of computed tomography findings in patients with bronchiolitis obliterans organizing pneumonia. AB - RATIONALE AND OBJECTIVES: The authors evaluated high-resolution computed tomography (HRCT) findings in 15 patients with biopsy-proven bronchiolitis obliterans organizing pneumonia (BOOP). Special attention was paid to lobar distribution to establish a predominant distribution pattern for this disease. SUBJECTS AND METHODS: Fifteen patients (average age, 61.8 years) with BOOP underwent computed tomography examinations. The examination technique included a slice thickness of 4 mm with a 4-mm interval; matrix size was 256 x 256 pixels. In all patients, additional HRCT images with a 2-mm slice thickness and a 10-mm interval (matrix, 512 x 512 pixels) were obtained. Interpretation included assessment of pulmonary, pleural, and mediastinal involvement. Special attention was paid to the distribution pattern of pulmonary abnormalities. RESULTS: High resolution computed tomography in all patients demonstrated areas of air space consolidation in a multifocal but peripheral distribution. The right lower lobe was involved in 60% of the patients, the left lower lobe in 53%, the middle and right upper lobes in 20%, and left upper lobe in 23%. Five patients had ground glass opacities in addition to the areas of air space consolidation, with the incidence in these patients being 100% in the right lower lobe. 80% in the left lower and middle lobes, 60% in the right upper lobe, and 20% in the left upper lobe. Nodules were found in two patients, and the left lower and middle lobes were affected in both. In one patient, the right lower lobe was affected. The interstitium was thickened in 66%, with axial involvement in 20%, septal thickening in 30%, and peripheral regions affected in 50%. Bronchiectasis was present in 60% of all patients studied, predominantly located in the lower lobes. CONCLUSION: Bronchiolitis obliterans organizing pneumonia presents a predominant peripheral, bilateral, and nonsegmental distribution, with the lower and middle lobes affected more than the upper lobes. PMID- 8724122 TI - Electrocardiographic changes in elderly patients during small bowel enema. AB - RATIONALE AND OBJECTIVES: Enteroclysis (small-bowel enema) involves the introduction of a large amount of fluid into the small bowel, through a tube, producing small bowel distention. A study was done to determine the incidence of any electrocardiographic changes during enteroclysis with Holter monitoring. METHODS: Continuous electrocardiographic monitoring and 12-lead electrocardiograms were performed in 30 elderly patients undergoing enteroclysis and in 30 control subjects undergoing routine chest, bone, and upper gastrointestinal small bowel follow-up studies. Two channel qualitative and quantitative electrocardiographic analysis was performed by a computerized nontriggered template system. Arrhythmias, change in cardiac axis, conduction defects, pauses, ST segment changes, and ectopics were sought. RESULTS: Increased sympathetic tone resulting in increased heart rate and transient atrial and ventricular ectopics was frequent during enteroclysis compared with the control group. In one patient ventricular tachycardia developed, and two patients had diminished heart rate, but this was attributed to preexisting heart disease and concurrent medication. CONCLUSION: Transient, nonhazardous cardiac arrhythmias are encountered during enteroclysis in elderly patients. These arrhythmias may be attributed to the preexisting heart disease, fear, and anxiety during intubation, or increased sympathetic tone from the enteric loop distention. PMID- 8724123 TI - Postsurgical outcome of patients with uncontrolled complex partial seizures and temporal lobe hypometabolism on 18FDG-positron emission tomography. AB - RATIONAL AND OBJECTIVE: The purpose of this study is to evaluate the relation between a focus of temporal lobe hypometabolism, including comparison between mesial and lateral asymmetry on fluorine-18-labeled-deoxyglucose-positron emission tomography (18FDG-PET) and surgical outcome in patients with uncontrolled partial seizures. METHODS: Case histories, electroencephalogram (EEG) findings, radiographic findings, and surgical outcome (36 +/- 11 months of follow-up) were reviewed in 38 consecutive patients who had a interictal 18FDG PET scan and subsequent temporal resection. RESULTS: Among the 36 patients who had a temporal lobe focus of hypometabolism (more than 15% asymmetry to contralateral side), 61% (22 of 36) became seizure-free, 33% (12 of 36) markedly improved and 6% (2 of 36) did not improve. The focus of hypometabolism on PET was in agreement with the epileptic focus on the noninvasive EEG in 30 of 36 patients and in 19 of the 22 patients who underwent an invasive EEG. The asymmetry index for the mesial temporal lobe was significantly higher in the group of patients who became seizure-free compared with the other patients. CONCLUSION: This study confirms that a focus of interictal temporal hypometabolism on PET is associated with marked improvement of seizure control after surgery in 94% (34 of 36) of the patients. Hypometabolism in the mesial temporal lobe appears to be associated with a seizure-free outcome. PMID- 8724124 TI - Differentiation of spontaneous canine breast tumors using dynamic magnetic resonance imaging with 24-Gadolinium-DTPA-cascade-polymer, a new blood-pool agent. Preliminary experience. AB - RATIONALE AND OBJECTIVES: The author assess the enhancement characteristics over time of spontaneous breast tumors in dogs comparing gadopentetate dimeglumine with a new blood-pool agent (24-gadolinium [Gd]-DTPA-cascade polymer). METHODS: Eighteen dogs with spontaneous breast tumors (5 carcinomas, 4 adenomas, and 9 benign mixed-tissue tumors) underwent dynamic magnetic resonance imaging after intravenous injection of gadopentetate dimeglumine and the blood-pool agent. Signal intensity time curves were followed up to 30 minutes after injection of both agents in the same animal. A nonlinear fitting routine enabled calculation of the delivery and clearance half lives of the contrast agent kinetics in each tumor. RESULTS: For gadopentetate dimeglumine, a fast signal increase was found immediately after intravenous injection, with a subsequent signal decay in all tumors. No difference was observed between the enhancement kinetics of different tumor types after gadopentetate dimeglumine application. Similar kinetics were found in benign lesions after injection of the blood-pool agent. However, in carcinomas the blood-pool agent displayed a slower delivery, delayed peak enhancement, and slower tumor tissue clearance or even a signal plateau of more than 30 minutes. CONCLUSIONS: Dynamic magnetic resonance imaging of breast neoplasms using a blood-pool agent may help to better differentiate between benign and malignant lesions because it demonstrates the enlarged interstitial space and increased capillary permeability in carcinomas. PMID- 8724125 TI - Vascular manifestations of small solitary pulmonary masses. Angiographic pathologic correlations and clinical significance. AB - RATIONALE AND OBJECTIVES: By performing pulmonary specimen angiographies, the authors attempted to determine the pathologic correlations of the vascular related radiologic manifestations within, at the edge of, and adjacent to the small solitary pulmonary masses, and to evaluate their usefulness in differentiating small bronchogenic carcinomas from tuberculomas. METHODS: A total of 29 resected lobe specimens with 1.5- to 3.5-cm solitary pulmonary masses, including 24 carcinomas and 5 tuberculomas, were studied prospectively with preoperative radiographs, postoperative specimen arteriographies (in 19 carcinomas and 5 tuberculomas), and venographies (in 5 carcinomas), 10- to 15 micrograms-thick whole-mount sections, and 5-micrograms-thick slices for the examination of angiographic-pathologic correlation. Another series of chest radiographs and conventional tomographs of 100 patients with 1- to 3-cm peripheral pulmonary masses, including 60 carcinomas and 40 tuberculomas, were reviewed retrospectively and analyzed with the chi-square test. RESULTS: Specimen angiographies showed the intralesion avascularity, small arterial speculation, and lobulation or notch at the mass margin with arterial compression, as well as vascular convergence to the mass, in both carcinoma and tuberculoma groups. The irregular arterial wall (79.2%) and venous dilation distal to the mass (100%) were found in the carcinoma group only. Microscopically, arterial or venous fibrous hyperplasia was observed in both carcinomas and tuberculomas, whereas the arterial erosion by tumor tissue and tumor emboli within the vessels were found in carcinomas only. The retrospective review of the 100 patients showed that two radiologic signs of the vascular convergence to the mass and the vascular dilation distal to the mass occurred at similar frequencies (12%-13%) between the carcinoma and tuberculoma groups. CONCLUSIONS: In addition to compression of vessels by tumor and vessel occlusion by tumor embolus, pulmonary vascular fibrotic hyperplasia can cause intramass avascularity. Small vessels running vertically into or from the mass margin can construct the spiculation sign of the tuberculomas. Any evidence of pulmonary vascular irregularity will indicate a bronchogenic carcinoma. The vascular convergence to the mass and the vascular dilation distal to the mass are not specific radiologic signs for small solitary bronchogenic carcinomas. PMID- 8724126 TI - Imaging of human colon cancer xenograft with gadolinium-texaphyrin. AB - RATIONALE AND OBJECTIVES: The authors explore the efficacy of gadolinium (Gd) texaphyrin (PCI-0101), an expanded porphyrin, as a contrast medium for magnetic resonance imaging of nude mice implanted with the human colon cancer xenograft LS174T. METHODS: Magnetic resonance images were obtained in six nude mice 7 to 8 days after implantation of LS174T cells in dorsal subcutaneous tissues. Spin-echo T1-weighted images were obtained at baseline and at 5, 15, and 60 minutes after injection of 10 or 20 mumols/kg of Gd-texaphyrin. Delayed images were obtained at 24 and 48 hours after injection. Region-of-interest measurements were taken of the tumors and of enhancing tumor rims at all time points. Percent enhancement was calculated and compared among the various time points. RESULTS: All tumors were enhanced after injection of Gd-texaphyrin. Heterogeneous patterns of enhancement were seen, with peak enhancement seen at the 15-minute time point; however, greater enhancement was seen at 48 hours than at 24 hours after Gd texaphyrin intravenous injection. CONCLUSIONS: Gadolinium-texaphyrin may prove to be a useful contrast medium for magnetic resonance imaging of tumors. PMID- 8724127 TI - Induction of sister chromatid exchange in the presence of gadolinium-DTPA and its reduction by dimethyl sulfoxide. AB - RATIONALE AND OBJECTIVES: The authors investigate the frequency of sister chromatid exchange (SCE) after the addition of gadolinium (Gd)-DTPA to venous blood samples. METHODS: Venous blood was obtained from nonsmokers. Samples were incubated with Gd-DTPA alone or in combination with mitomycin C, cytarabine, and dimethyl sulfoxide (DMSO), and then evaluated for SCEs. RESULTS: The frequency of SCE increased with the concentration of Gd-DTPA and as each chemotherapeutic agent was added. Sister chromatid exchange frequencies were lower when the blood was treated with a combination of Gd-DTPA and DMSO compared with Gd-DTPA alone. DISCUSSION: The increase in frequency of SCE seen after the addition of Gd-DTPA was decreased by the addition of DMSO, indicating the production of hydroxyl radicals. The effect likely is dissociation-related. PMID- 8724128 TI - Carboxymethyl-dextran-gadolinium-DTPA as a blood-pool contrast agent for magnetic resonance angiography. Experimental study in rabbits. AB - RATIONALE AND OBJECTIVES: The authors evaluate the efficiency of various doses of a paramagnetic macromolecular contrast agent, a gadolinium (Gd)-DTPA-dextran conjugate, as a blood-pool contrast media, in a transverse three-dimensional time of-flight (TOF) magnetic resonance (MR) angiography sequence of the abdominal aorta in rabbits. METHODS: Imaging experiments were performed on a 1.5-T magnet, using a transverse three-dimensional TOF tilted optimized nonsaturating excitation (TONE) sequence. The macromolecular contrast media used was a carboxymethyl-dextran-Gd-DTPA (CMD-Gd-DTPA). Different concentrations of CMD-Gd DTPA (0.005, 0.01, 0.03, 0.05 mmol Gd/kg) were evaluated. A comparative study using Gd-DOTA (0.01 and 0.1 mmol/kg) was performed. A visual analysis based on the gain in the visualized length of small arteries (renal arteries), and a quantitative analysis based on the percent contrast enhancement of the aorta plotted against distance in the slab from the top edge of the acquisition volume were obtained. RESULTS: A signal-to-noise ratio enhancement of the distal part of the aorta and an improvement in the visualized length of the renal arteries were noted for concentrations of CMD-Gd-DTPA ranging form 0.01 to 0.05 mmol Gd/kg. Venous enhancement was noted for concentrations greater than 0.01 mmol Gd/kg when using CMD-Gd-DTPA or Gd-DOTA. CONCLUSION: Carboxymethyl-dextran-Gd-DTPA reduced, in part, the saturation effect in a three-dimensional transverse TOF TONE MR angiography in rabbits. To prevent venous enhancement, observed with the higher concentrations used in this study, a decrease in the polydispersity of the polymer should be a goal in the future. Rapid extravasation of the low-molecular weight fraction of the polymer could explain the venous enhancement. PMID- 8724129 TI - The use of gadolinium-BOPTA on magnetic resonance imaging in brain infection. AB - RATIONALE AND OBJECTIVES: The use of gadolinium (Gd)-BOPTA as a magnetic resonance contrast agent for central nervous system disease was studied in a canine brain abscess model. METHODS: A Streptococcus faecalis brain abscess was evaluated in five dogs at 1.5T. Imaging was performed during the late cerebritis stage, at 5 to 7 days after surgery. Magnetic resonance scans were acquired before and at 1, 5, 15, 30, 45, and 60 minutes after contrast administration, using a dose of 0.1 mmol/kg. Scans also were acquired both before and after contrast injection with the implementation of magnetization transfer. RESULTS: Lesion enhancement, quantified by region-of-interest measurement, peaked at 5 minutes after contrast injection. Both the increase in lesion enhancement from 1 to 5 minutes after injection and the decrease from 5 to 15 minutes after injection, although small, were statistically significant (P < 0.004 and P < 0.03, respectively). The application of magnetization transfer improved lesion enhancement, as measured by signal difference/noise, by 39%. This result also was statistically significant (P < 0.001). CONCLUSIONS: In intraparenchymal brain infection, Gd-BOPTA provides effective lesion enhancement when used at a dose of 0.1 mmol/kg. Further research is needed to compare the magnitude of enhancement achieved with Gd-BOPTA, which has weak protein binding and both hepatobiliary and renal excretion, with that with Gd chelates, which have pure renal excretion. PMID- 8724130 TI - Consistency of rotator-cuff calcifications. Observations on plain radiography, sonography, computed tomography, and at needle treatment. AB - RATIONALE AND OBJECTIVES: The author analyzes findings of consistency of rotator cuff calcifications found at ultrasound (US)-guided needle treatment compared with findings of plain radiography, US, and computed tomography (CT). METHODS: Twenty patients had rotator-cuff calcifications (mean diameter, 1.5 cm; range, 1.1-2.6 cm) resistant to conservative therapy. At needle treatments, the consistency was assessed as hard or soft (slurry calcific deposit). In each imaging examination, the calcifications were divided into two groups. In radiographs, calcifications were divided into "well-defined" and "ill-defined." In US, they were divided into calcifications with acoustic shadow and calcifications with a faint shadow or no shadow. On CT, they were divided into homogeneous or nonhomogeneous calcifications. At CT, the density of the calcifications also was determined. The findings of consistency obtained at needle treatments were compared with the findings of plain radiography, US, and CT. RESULTS: At needle treatments, 45% (9 of 20) of the calcifications were soft or nearly liquid, and 55% (11 of 20) were hard. On plain radiographs, 67% (6 of 9) were as soft and 64% (7 of 11) as hard. On sonograms, 77% (7 of 9) were soft and 82% (9 of 11) were hard. On CT images, 77% (7 of 9) were soft and 91% (10 of 11) were hard; CT attenuation values were 77% (7 of 9) and 91% (10 of 11), respectively. CONCLUSIONS: Ultrasound and CT were reliable in predicting the consistency of rotator-cuff calcifications. Computed tomography attenuation values were the most accurate, and plain radiographs were the least accurate. PMID- 8724131 TI - Limitations of hyperselective intraarterial injections for the treatment of hepatocellular carcinoma. AB - The authors report the case of a cirrhotic patient with a multinodular hepatocellular carcinoma. Two nodules were located in the right liver lobe and a minute nodule was in the left lobe. Because of poor liver function, two injections of iodine-131-labeled Lipiodol were delivered in the right hepatic artery to protect the left lobe. The efficacy was obvious in the treated areas, but the minute lesion enlarged dramatically and was responsible for the patient's death. PMID- 8724132 TI - Renaturation and ligand blotting of the major subunit of the rat asialoglycoprotein receptor after denaturing polyacrylamide gel electrophoresis. AB - Rat hepatic asialoglycoprotein receptors (ASGP-Rs) bind terminal clustered galactosyl or N-acetylgalactosaminyl residues with high affinity. The affinity purified ASGP-R consists of three subunits designated RHL1, RHL2, and RHL3. The ligand-binding activity of individual subunits was investigated by ligand blotting, after separation of subunits by SDS-PAGE under nonreducing conditions, electrotransfer to nitrocellulose, and incubation with 125I-asialo-orosomucoid (ASOR). No ligand-binding to any subunits could be detected when proteins such as BSA, casein, gelatin, or fat-free dry milk were used as blocking agents. However, subsequent incubation of BSA-blocked nitrocellulose blots with some nonionic detergents resulted in renaturation of RHL1. 125I-ASOR-binding to RHL2 or RHL3 was weaker and could be detected only after longer exposure. Similarly, direct use of detergents such as Tween 20, Nonidet P-40, or Triton X-100 as blocking agents also preserved the ASOR-binding activity of RHL1. Ionic detergents tested did not show any ability to renature the ligand-binding activity of RHL subunits. Among nonionic detergents tested, Tween 20, Tween 85, Lubrol PX, Nonidet P-40, and Triton X-100 were more effective than Tween 40, Tween 65, Tween 80, or Brij 35, whereas SPAN, digitonin, or octyl-glucoside showed no effect. Weak 125I-ASOR binding to RHL2 or RHL3 could be detected only when the Tween series or Lubrol PX were used. Incubation of blots with dithiothreitol caused a dose-dependent loss of binding activity. The carbohydrate recognition domain (CRD) of RHL1, isolated after subtilisin digestion of ASGP-R bound to ASOR-Sepharose, retained ligand binding activity as assessed by its binding to ASOR-Sepharose and by ligand blotting. 125I-ASOR binding to electroblotted CRD after SDS-PAGE was also dependent on the presence of nonionic detergents. We conclude that restoration of ligand-binding activity of RHL1 after SDS-PAGE by some nonionic detergents is not dependent on the presence of the cytoplasmic, transmembrane, or stalk domains of this subunit. PMID- 8724133 TI - Distinctive ganglioside patterns revealed by anti-ganglioside antibody binding to differentiating CG-4 oligodendrocytes. AB - Oligodendrocytes are central nervous system glial cells responsible for myelination of neuronal axons. During brain development oligodendrocyte progenitor cells progress through a series of morphologically and immunohistochemically distinct differentiation steps leading to mature myelin producing oligodendrocytes. Much of this same differentiation sequence is expressed in vitro by primary oligodendrocyte progenitor cells, and by the clonal progenitor cell line CG-4. We report the use of highly specific monoclonal antibodies against GM1, GD1a, GD1b, GT1b, and GQ1b to determine major brain ganglioside expression and morphological distribution during CG-4 differentiation in vitro. Prominent anti-GD1b antibody stained defined a highly arborized intermediate stage of oligodendrocyte differentiation. In contrast, anti-GT1b antibody bound to discrete patches on the cell bodies of early progenitor cells and more mature oligodendrocytes, and to sites of progenitor arborization. The other anti-ganglioside antibodies tested did not bind above background levels. Cells with anti-GD1b antibody binding and morphology similar to those in differentiating CG-4 cells were detected in rat brain primary cell cultures enriched in oligodendrocyte precursors. The remarkably distinctive ganglioside immunoreactivity on differentiating oligodendrocytes suggests the possibility of a functional role for their surface expression. PMID- 8724134 TI - A spectrophotometric assay for alpha-mannosidase activity. AB - A simple and versatile spectrophotometric assay for alpha-mannosidase activity, which can be used with unlabelled natural substrate or synthetic substrates, was developed. The reducing mannose released from the substrate by the enzyme is quantitated using glucose oxidase, peroxidase and o-dianisidine. Using recombinant alpha 1,2-mannosidase obtained from Saccharomyces cerevisiae and Man9, GlcNAc, the spectrophotometric assay yielded values of 0.3 mM for Km and 15 mU/microgram for V(max), which are comparable to those obtained using the traditional radiochemical assay. The assay was also used to evaluate some alternative oligosaccharides as substrates for the enzyme. Man5-O(CH2)8-COOCH3 shows potential as an alternative synthetic substrate as the enzyme retained its specificity for a single alpha 1,2-mannose residue. Kinetic results suggest that the lower 1,3 linked arm of Man9GlcNAc is more critically involved in substrate recognition than the upper 1,6 linked arm. PMID- 8724135 TI - Transcription of the beta-galactoside alpha 2,6-sialyltransferase gene in B lymphocytes is directed by a separate and distinct promoter. AB - A single human gene, SIAT1, encodes the beta-galactoside alpha 2,6 sialyltransferase from which multiple mRNA isoforms are generated. In rat, expression of the hepatic mRNA isoform (Form 1) has been defined with respect to the transcriptional initiation site and promoter region. We show here that a similar hepatic SIAT1 mRNA isoform exists in human. Another human mRNA isoform, a mature B-cell-specific mRNA isoform (Form 2), was previously reported. Here, we used 5'-RACE and S1 nuclease protection analysis to define the 5'-untranslated region of Form 2 human SIAT1 mRNA. We demonstrate conclusively that Form 2 mRNA is initiated from a point completely distinct from that of Form 1 mRNA. A number of cis-acting regulatory elements residing immediately 5'of the Form 2 initiation site includes AP-1, AP-2, NF-kappa B, NF-IL6, C/EBP, and CREB. A TATAA box is also present 29 bp 5' of the transcriptional initiation site. CAT reporter gene expression from serially-truncated segments of the 5'-flanking region of the Form 2 initiation site indicates that the segment between -784 and +125 was sufficient to promote high level CAT expression in Louckes, a mature B-cell line. The 5' flanking region to the human Form 1 initiation site is competent in expression of CAT upon transfection of the fusion construct into HepG2, a human hepatoma cell line. Cellular specificity of expression is apparently retained. Louckes cells expressed CAT efficiently from Form 2 promoter but only marginally from the Form 1 promoter. In contrast, CAT expression from Form 1 promoter is more efficient than from the Form 2 promoter in HepG2 cells. PMID- 8724136 TI - Comparison of NMR and molecular modeling results for a rigid and a flexible oligosaccharide. AB - Three-bond heteronuclear coupling constants (3JCH) are extremely useful in describing flexible models for oligosaccharides. We show that antiphase methods for measuring 3JCH in oligosaccharides have limited reliability but that the coupling constants can be reliably measured in natural abundance by quantitative J-correlation methods. Interpretation of 3JCH data for a pentasaccharide (lacto-N fuco-pentaose 2) from human milk are consistent with a rigid model for the Lewis(a) trisaccharide epitope but for an antigenic tetrasaccharide fragment from the cell wall polysaccharide of viridans streptococci, 3JCH data imply a considerably more flexible model. Nuclear Overhauser effect (NOE) data are reported for a heptasaccharide repeating unit isolated from the cell wall polysaccharide of Streptococcus gordonii 38. The results for a tetrasaccharide fragment are similar to data reported for the same fragment in the cell wall polysaccharide from S.mitis J22. This result implies a similar conformation for the tetrasaccharide fragment in the polysaccharide and in the heptasaccharide and also implies that anisotropy of motion is not significant in the interpretation of the nuclear Overhauser effects in the polysaccharide. Interpretation of the NOE results for the tetrasaccharide fragment, like the 3JCH data, implies a flexible model with three conformations in fast exchange. The results of the two experimental techniques are combined with molecular modeling results including molecular dynamics simulation to provide a clear delineation between flexible and rigid oligosaccharide epitopes. The blood group Lewis(a) trisaccharide antigenic determinant is highly restricted in its motions by steric interactions while the antigenic tetrasaccharide fragment of the S.gordonii 38 heptasaccharide is considerably more mobile. We propose that some branched oligosaccharides are relatively rigid and some are flexible depending on subtle details of the linkages. PMID- 8724137 TI - Unique alpha 2, 8-polysialylated glycoproteins in breast cancer and leukemia cells. AB - The N-linked oligosaccharides of neural cell adhesion molecule and the rat brain voltage-dependent sodium channel alpha subunit are specifically modified by alpha 2, 8-polysialic acid chains. Until now, this carbohydrate modification has been observed only on these two proteins in mammalian cells. We have identified 180 260 kDa proteins in RBL rat basophilic leukemia cells and MCF7 human breast cancer cells that are modified by alpha 2, 8-polysialylated oligosaccharides. Immunofluorescence microscopy and Northern analysis confirmed that these proteins are neither the neural cell adhesion molecule nor the sodium channel alpha subunit. The presence of authentic alpha 2, 8-polysialic acid on the basophilic leukemia and breast cancer proteins was confirmed by the elimination of anti polysialic acid antibody staining after treatment with the alpha 2, 8-polysialic acid-specific endo-N-acetylneuraminidase. The failure of peptide N-glycosidase F to completely remove alpha 2, 8-polysialic acid bearing oligosaccharides from the RBL protein, and the sensitivity of these oligosaccharides to beta-elimination, suggests that alpha 2, 8-polysialic acid may be found on O-linked oligosaccharides. This identification of new alpha 2, 8-polysialylated proteins in RBL basophilic leukemia and MCF7 breast cancer cells suggests that alpha 2, 8 polysialylation of glycoproteins may be more widespread than originally believed, especially in cancer cells. PMID- 8724138 TI - Oxidized low density lipoprotein stimulates aortic smooth muscle cell proliferation. AB - We have investigated the effects of oxidized low density lipoproteins (Ox-LDL) on aortic smooth muscle cell (SMC) proliferation and the biosynthesis of glycosphingolipids. We found that Ox-LDL exerted a concentration, time, and temperature dependent alteration of cell proliferation and the biosynthesis of lactosylceramide. At low concentrations (5-10 micrograms/ml medium) Ox-LDL stimulated cell proliferation measured by an increase in the incorporation of [3H]-thymidine in cells and the synthesis of lactosylceramide, but not glucosylceramide synthesis. Oxidized LDL exerted a threefold increase in the incorporation of [3H]-galactose and [3H]-serine in lactosylceramide. The activity of lactosylceramide synthetase; UDP-galactose glucosylceramide beta 1 --> 4 galactosyltransferase (GalT-2), but not glucosylceramide synthetase (GlcT-1) was stimulated by Ox-LDL. On the other hand, LDL suppressed the activity of GalT-2 in these cells. When cells were preincubated with antibody against Ox-LDL or GalT-2 it compromised the Ox-LDL mediated stimulated in cell proliferation and GalT-2 activity. Similarly, D-PDMP an inhibitor of GalT-2 compromised the Ox-LDL mediated effects in cells. In contrast, L-PDMP further stimulated the Ox-LDL mediated cell proliferation and GalT-2 activity. However, preincubation of cells with preimmune rabbit serum IgG failed to abrogate Ox-LDL mediated stimulation in cell proliferation and GalT-2 activity. In sum, we found that Ox-LDL stimulated aortic smooth muscle cell proliferation in culture. This effect resulted from Ox LDL mediated activation of GalT-2 that produced lactosylceramide. Lactosylceramide in turn, contributed to cell proliferation. Such correlations are supportive of the notion that GalT-2 action mediates the signal transduction of Ox-LDL contributing to cell proliferation. PMID- 8724139 TI - Defective threonine-linked glycosylation of human insulin-like growth factor in mutants of the yeast Saccharomyces cerevisiae. AB - Mutants of the yeast Saccharomyces cerevisiae were identified, in which O glycosylation at threonine 29 of a heterologous protein, human insulin-like growth factor (hIGF-1), is defective. In mutant M195, O-glycosylation of hIGF-1, but not of yeast proteins chitinase and a-agglutinin, was reduced; in mutant M577 yeast proteins were affected besides hIGF-1. The mutations of M195 and M577 did not affect viability and could not be complemented by the PMT1 or PMT2 genes. The mutant phenotype of strain M195 was reconstituted in an in vitro system, in which a hIGF-1-derived peptide encompassing residues 24-34 was not used as acceptor for mannosylation, while unrelated peptides were glycosylated at wild-type levels. hIGF-1 glycosylation was drastically reduced in pmt1 disruptants and to a lesser extent in pmt2 disruptants, suggesting interaction between the PMT gene products and components mutated in M195 and M577 cells. The results suggest that mutations may only affect O-glycosylation of a specific subset of secreted proteins in yeast. PMID- 8724140 TI - Forssman disaccharide is the specific ligand of a galectin from the sponge Geodia cydonium but does not mediate its binding to nuclear protein np56. AB - The galectin from Geodia cydonium (GCA) had previously been shown to be involved in regulatory mechanisms of cell sorting and adhesion during reaggregation of allogeneic sponge cells. In this contribution the binding specificity of GCA was established to be GalNAc alpha 1-3GalNAc beta as structural component of Forssman pentasaccharide. Crossreactivities of terminal structural elements were revealed in the order GalNAc alpha 1-3GalNAc beta > GalNAc alpha 1-3(Fuc alpha 1-2)Gal beta >> Gal beta 1-3GlcNAc beta > Gal beta 1-4Glc. Lectin binding to the Forssman antigen (Ki range 10(-7)) or to blood group A-trisaccharide exceeded that to lactose (Ki range 10(-3)/10(-2)) by three to four orders of magnitude. Cytochemical staining of eukaryotic cells on the light and electron microscopic level revealed lectin binding in the cytosol and in the nucleus (nucleoli), which was inhibitable with the soluble high affinity ligands. The nuclear binding of GCA could be ascribed to affinity-isolated 56 kDa protein (np56) in the nucleoplasm and was shown to be mediated by the peptide conformation of the ligand. Although GCA-np56 interaction was inhibitable with Forssman glycolipid or globopentaose, the carbohydrate binding site of the lectin is not involved due to the lack of competition by Forssman-specific lectins HPA or DBA. Since anti-CBP70 was immunologically cross-reactive to np56, it is concluded that the galectin GCA binds to np56 via similar mechanisms as reported previously for the interaction of CBP-35 (galectin-3) and CBP-70. Thus, GCA resembles galectin-3 in its binding characteristics but is likewise related to galectin-1 by sequence homology of its primary structure and by the molecular mass of its subunits. PMID- 8724141 TI - Retention of Saccharomyces cerevisiae cell wall proteins through a phosphodiester linked beta-1,3-/beta-1,6-glucan heteropolymer. AB - Yeast cell wall proteins, including Cwp1p and alpha-agglutinin, could be released by treating the cell wall with either beta-1,3-or beta-1,6-glucanases, indicating that both polymers are involved in anchoring cell wall proteins. It was shown immunologically that both beta-1,3- and beta-1,6-glucan were linked to yeast cell wall proteins, including Cwp1p and alpha-agglutinin. It was further shown that beta-1,3-glucan was linked to the wall protein through a beta-1,6-glucan moiety. The beta-1,6-glucan moiety could be removed from Cwp1p and other cell wall proteins by cleaving phosphodiester bridges either enzymatically using phosphodiesterases or chemically using ice-cold aqueous hydrofluoric acid. These observations are consistent with the notion that cell wall proteins in Saccharomyces cerevisiae are linked to a beta-1,3-/beta-1,6-glucan heteropolymer through a phosphodiester linkage and that this polymer is responsible for anchoring cell wall proteins. It is proposed that this polymer is identical to the alkali-soluble beta-1,3-/beta-1,6-glucan heteropolymer characterized by Fleet and Manners (1976, 1977). PMID- 8724142 TI - Effects of exogenous gangliosides on intracellular Ca2+ mobilization and functional responses in human platelets. AB - Gangliosides, highly expressed in the outer leaflet of plasma membranes, mediate a variety of biological processes, including cell-cell and cell-matrix interactions. We examined the effects of exogenous gangliosides on intracellular Ca2+ mobilization and functional responses in human platelets. Gangliosides (GM3 and GM1) induced rapid and reversible elevation of intracellular Ca2+ in fura2 loaded platelets in a concentration-dependent manner. The Ca(2+)-mobilizing effect of gangliosides was not mimicked by deN-acetyl-GM3, lactosylceramide, or free sialic acid, suggesting that structural integrity as ganglioside is essential for this effect. GM3 and GM1 also induced platelet shape change by themselves and elicited aggregation in combination with epinephrine. Our observations suggest the involvement of ganglioside-activated platelets in atherosclerosis, in view of the high observed ganglioside levels in atherosclerotic lesions of human aorta. PMID- 8724143 TI - Chemical modifications of heparin that diminish its anticoagulant but preserve its heparanase-inhibitory, angiostatic, anti-tumor and anti-metastatic properties. AB - Structural features of heparin potentially important for heparanase-inhibitory activity were examined by measuring the ability of heparin derivatives to affect the degradation of [3H]acetylated heparan sulphate by tumor cell heparanases. IC50 values were determined using an assay which distinguished degraded from undegraded substrate by precipitation of the latter with cetylpyridinium chloride (CPC). Removal of heparin's 2-O-sulphate and 3-O-sulphate groups enhanced heparanase-inhibitory activity (50%). Removal of its carboxyl groups slightly lowered the activity (18%), while combining the treatments abolished the activity. At least one negative charge on the iduronic acid/idose moiety, therefore, is necessary for heparanase-inhibitory activity. Replacing heparin's N sulphate groups with N-acetyl groups reduced its activity (37%). Comparing this heparin derivative with 2,3-O-desulphated heparin, the placement of sulphate groups appears important for activity since the two structures have similar nominal linear charge density. In addition, unsubstituted uronic acids are nonessential for inhibition since their modification (periodate oxidation/borohydride-reduction) enhanced rather than reduced heparanase inhibitory activity. The most effective heparanase inhibitors (2,3-O-desulphated heparin, and [periodate-oxidized, borohydride-reduced] heparin) were tested in the chick chorioallantoic membrane (CAM) bioassay for anti-angiogenic activity and found to be at least as efficacious as heparin. 2,3-O-desulphated heparin also significantly decreased the tumor growth of a subcutaneous human pancreatic (Ca-Pan-2) adenocarcinoma in nude mice and prolonged the survival times of C57BL/6N mice in a B16-F10 melanoma experimental lung metastasis assay. PMID- 8724144 TI - Selective ganglioside desialylation in the plasma membrane of human neuroblastoma cells. AB - Gangliosides of the plasma membrane are important modulators of cellular functions. Previous work from our laboratory had suggested that a plasma membrane sialidase was involved in growth control and differentiation in cultured human neuroblastoma cells (SK-N-MC), but its substrates had remained obscure. We now performed sialidase specificity studies in subcellular fractions and found ganglioside GM3 desialylating activity in presence of Triton X-100 to be associated with the plasma membrane, but absent in lysosomes. This Triton activated plasma membrane enzyme desialylated also gangliosides GD1a, GD1b, and GT1b, thereby forming GM1; cleavage of GM1 and GM2, however, was not observed. Sialidase activity towards the glycoprotein fetuin with modified C-7 sialic acids and towards 4-methylumbelliferyl neuraminate was solely found in lysosomal, but not in plasma membrane fractions. The role of the plasma membrane sialidase in gangliosides desialylation of living cells was examined by following the fate of [3H]galactose-labelled individual gangliosides in pulse-chase experiments in absence and presence of the extracellular sialidase inhibitor 2-deoxy-2,3-dehydro N-acetylneuraminic acid. When the plasma membrane sialidase was inhibited, radioactivity of all gangliosides chased at the same rate. In the absence of inhibitor, GM3, GD1a, GD1b, GD2, GD3 and GT1b were degraded at a considerably faster rate in confluent cultures, whereas the GM1-pool seemed to be filled by the desialylation of higher gangliosides. The results thus suggest that the plasma membrane sialidase causes selective ganglioside desialylation, and that such surface glycolipid modification triggers growth control and differentiation in human neuroblastoma cells. PMID- 8724145 TI - Thirty-six month follow-up of excimer laser photorefractive keratectomy for myopia. AB - BACKGROUND AND OBJECTIVE: The aim of the study was to evaluate the refractive outcome 36 months following excimer laser photorefractive keratectomy (PRK) performed on myopic eyes. We analyzed the refractive stability between 24 and 36 months. PATIENTS AND METHODS: PRK was performed on 457 eyes with a pre-operative refraction ranging from -1.25 to -7.50 diopters (D). Ablation zone diameters of 4.3 to 4.5 mm were used. The follow up time was 36 months. RESULTS: The mean refraction 36 months following surgery was -0.22 to 0.75 D for the whole group, which was not significantly different from the mean at 24 months (-0.27 +0.74D). If we analyze the subgroups at 36 months, we find a significantly better refractive outcome in the low myopia group (up to -2.90D) compared with those with higher myopia. At 36 months, 91% of the 456 eyes had an uncorrected visual acuity of at least 20/40, and 88% of the eyes were within 1.00 D of emmetropia. CONCLUSION: These results show stability between 24 and 36 months. PMID- 8724146 TI - Corneal asphericity following excimer laser photorefractive keratectomy. Summit PRK Topography Study Group. AB - BACKGROUND AND OBJECTIVE: To analyze corneal asphericity following excimer laser photorefractive keratectomy (PRK) and its influence on clinical outcomes. PATIENTS AND METHODS: A computer program (Holladay Diagnostic Summary, EyeSys Laboratories, Houston, TX) was used to qualitatively and quantitatively analyze the corneal asphericity of 132 patients 1 year following PRK for correction of myopia. Color maps depicting actual corneal asphericity as compared to the normal expected asphericity were reviewed, and quantitative values of asphericity were evaluated for associations with clinical outcomes of uncorrected visual acuity and spectacle corrected visual acuity, achieved refractive correction, a subjective glare/halo index, and subjective patient satisfaction, as well as standard corneal topography patterns and optical zone decentration following PRK. RESULTS: Following PRK, all corneas exhibited a positive central asphericity, changing from a prolate (negative asphericity) to an oblate optical contour. There was a trend toward higher positive asphericity measurements with improving spectacle corrected visual acuity which was not statistically significant; such a relationship was not found with uncorrected visual acuity. A significant association was found between greater achieved refractive correction and increased postoperative positive asphericity. No association was found between postoperative asphericity and the glare/halo index, subjective patient satisfaction, topography pattern, or optical zone decentration. CONCLUSION: Asphericity may be a useful quantitative descriptor of corneal optical contour following PRK. Greater positive central corneal asphericity is found with greater degrees of refractive correction. Further understanding of both the pre- and post operative corneal contour and the consequent optical effects should aid in a better understanding of the optical outcomes of PRK. PMID- 8724147 TI - Analysis of photorefractive keratectomy patients who have not had PRK in their second eye. AB - BACKGROUND AND OBJECTIVE: Excimer laser photorefractive keratectomy (PRK) is performed at the Singapore National Eye Centre (SNEC) with a minimum period of 3 months between first and second eye treatment. During this period, iatrogenic anisometropia may occur, which can lead to significant visual disability. We analyzed the reasons why some patients delayed or did not receive PRK treatment to their second eye. The reasons for originally electing PRK, and the expectations these patients had, were also studied. PATIENTS AND METHODS: Between January 1992 and September 1993, 341 patients underwent PRK at SNEC. Of these, 86 (25.2%) did not receive PRK to their fellow eye within 1 year of follow-up and were recruited into the study. Data were collected retrospectively using a standardized questionnaire, and objective clinical data were obtained from the case records. Sixty-eight patients (79%) responded. RESULTS: Fifty-one patients (14.9%) had not undergone PRK at the conclusion of the study (mean follow-up = 31 months). Of these, 36 were due to dissatisfaction with the results of the procedure. Symptoms relating to hypermetropia (15 patients) and symptoms of glare and halos were the main reasons which discouraged patients from receiving PRK in their fellow eye. Seventeen of the 51 patients (33.3%) who did not have PRK performed in the second eye were overcorrected to hypermetropia, as opposed to 9 out of 255 patients (3.5%) who had PRK in the second eye (P < 0.001, chi-square test). Seventy-five percent of the patients who did not undergo PRK in the second eye expected postlaser unaided vision to be equal to prelaser best corrected vision; however, only 23% of these patients achieved this. CONCLUSION: Hypermetropia, glare and halos are significant causes of patient dissatisfaction after PRK. Patients who request PRK should be advised against harboring unrealistically high expectations for the procedure. PMID- 8724148 TI - Myopic regression after photorefractive keratectomy. AB - BACKGROUND AND OBJECTIVE: Myopic photorefractive keratectomy (PRK) using the 193 nm excimer laser is an effective and precise surgical procedure to correct myopia, but not without complications, such as corneal haze, over- or undercorrection, halo, glare, and myopic regression. Among these, myopic regression, which can reduce uncorrected visual acuity, is the most common and progressive long-term complication. PATIENTS AND METHODS: We evaluated 228 eyes that received successful myopic PRK with the Summit excimer laser (ExciMed UV 200LA, 5.0 mm optical zone) and followed up more than three years at Kangnam St. Mary's hospital. Two hundred twenty eight eyes were divided into 3 groups according to the degree of preoperative myopia and correction: Group I, 79 eyes of moderate myopia (-2.25 to-6.75 diopters [D]) with full correction; Group II, 110 eyes of high myopia (-7.00 to -12.50 D) with full correction by two-zone ablation; Group III, 29 eyes of high myopia (-7.00 to -12.00 D) with partial correction of 6.00 D. Retrospective analysis of the data was performed with statistical evaluation of the incidence of myopic regression, regression formula, coefficient of parameters, and cumulative hazard of regression in each group. RESULTS: An incidence of myopic regression greater than 1.00 D was noted in 40.5%, 83.3%, and 18.2% of patients in Group I, II, and III, respectively. The regression formula obtained from the repeated measure ANOVA was Y = 2.13 -0.47 InX, Y = 3.02 -0.82 InX, and Y = 3.69 -0.50 InX, in Group I, II, and III, respectively. The amount of correction was a statistically significant correlation factor to the myopic regression (Pearson's correlation coefficient = 0.78, P < 0.001). The amount of preoperative myopia may act as a negative correlation factor to the myopic regression. According to the Kaplan-Meyer survival table, the cumulative correlation hazard was significantly higher in high myopia with full correction than in moderate myopia with full correction. CONCLUSION: Our results indicated that the incidence and amount of myopic regression have a tendency to be dependent on the amount of correction, but may be reduced as the preoperative myopia increases, if the amount of correction is the same. A longer term and more detailed study of PRK using a 6.0 mm optical zone will be needed to determine how the amount of preoperative myopia and would profiles affect the myopic regression. PMID- 8724149 TI - Early visual recovery after excimer laser surgery for myopia: the Melbourne OmniMed results. AB - BACKGROUND AND OBJECTIVE: Although the refractive outcome can change for months after photorefractive keratectomy (PRK), early visual results are of particular importance during pre-operative counseling of prospective PRK candidates. The expected vision in the first week post-operatively, following 6.0 mm or larger zone photoablation for myopia with the Summit OmniMed (Apex) laser, has not been analyzed in the literature to date. This study is intended to provide an indication of the early visual acuity that may be experienced by a patient undergoing PRK for myopia. PATIENTS AND METHODS: The visual acuity recorded for 123 consecutive patients, (79 low myopes [-1.00 to -5.90 diopters (D)], 44 high extreme myopes [-6.00 to -19.00 D]), one week after they had undergone PRK, was analyzed. All patients included in the study had undergone PRK with the Summit OmniMed (Apex) excimer laser, leaving small degrees of astigmatism uncorrected. RESULTS: One week after PRK, uncorrected visual acuity was better for low myopes than high myopes. Of the low myopes, 83.0% were 20/40 or better compared with high myopes of whom 61.4% were 20/40 or better. CONCLUSION: Patients can be counseled that functional vision will be present in the period immediately after re-epithelialization. PMID- 8724150 TI - Photorefractive keratectomy: the Sankara Nethralaya experience. AB - BACKGROUND AND OBJECTIVE: Excimer laser photorefractive keratectomy (PRK) is quickly establishing itself as a dependable treatment modality for the correction of myopia. We review in this report our experience with the Summit OmniMed UV200 excimer laser (Summit Technology, Inc., Waltham, MA) in the treatment of low to high degrees of myopia. PATIENTS AND METHODS: We retrospectively analyzed the results of excimer laser PRK in 139 eyes of 127 patients treated between November 1993 and August 1994. RESULTS: The study population was comprised of 75 males and 52 females with a mean ago of 26.9 +/- 6.21 years (range, 19 to 52 years). All patients were followed for at least six months and the mean follow-up was 11.7 +/ 3.63 months (range, 6 to 22 months). Mean pre-operative myopia was -8.23 +/- 3.32 diopters (D) (-2.63 to -19.50 D). Of 139 eyes, 101 (72.7%) had a pre operative myopia greater than -6.00 D; and 32 (23.0%) had a pre-operative myopia greater than -10.00 D. At last follow-up, 73 (52.5%) eyes were within +/-1.00 D of emmetropia, 92 (66.2%) eyes had an uncorrected visual acuity of > or = 6/12, and 12 (8.6%) eyes lost 2 or more lines of spectacle corrected visual acuity. Severe corneal haze (grade 3 to 4) was seen in 20 (14.4%) eyes at last follow-up. Complications encountered in this series included steroid response in 15 (10.8%) eyes, and reactivation of herpes simplex keratitis in one (0.7%) eye. Statistical analysis reveals that the only risk factor independently associated with regression > or = 2.00 D following PRK is the occurrence of severe corneal haze (grade 3 and 4). CONCLUSION: Excimer laser PRK appears to be a safe and predictable method for the treatment of myopia < 6.00 D. With higher degrees of myopia the results appeared to be more guarded, mainly due to the healing pattern of the cornea following single zone myopic ablations. PMID- 8724151 TI - Excimer laser photorefractive keratectomy: one year follow-up. AB - BACKGROUND AND OBJECTIVE: To confirm the effectiveness of the 193 nm excimer laser to correct low myopia (1.00 to 6.00 D), moderate myopia (6.00 to 10.00 D), and high myopia (10.00 D and above). PATIENTS AND METHODS: One hundred eighty-one eyes of 114 patients underwent excimer laser photorefractive keratectomy (PRK). The minimum follow-up was 12 months. RESULTS: After one year, 96% of the eyes in the low myopia group, 69% in the moderate myopia group and 29% in the high myopia group achieved uncorrected visual acuity of 20/40 or better with one treatment using a single application with a 5 mm optic zone. There were no significant major complications. CONCLUSION: PRK is a safe, effective and relatively accurate procedure to correct low to moderate myopia. PMID- 8724152 TI - Evaluation of three different approaches to perform excimer laser photorefractive keratectomy for myopia. AB - BACKGROUND AND OBJECTIVE: The aim of this study was to evaluate the results of myopic excimer laser photorefractive keratectomy (PRK) performed using three different ablation approaches. PATIENTS AND METHODS: Using the Apex excimer laser (Summit Technology, Inc., Waltham MA), we have treated 75 eyes. Twenty-five of them were treated by a single ablation zone (SA, 6.5 mm diameter) to correct myopia of -4.50 +/- 2.10 D (mean +/- SD); 25 eyes underwent PRK for the correction of myopia of -4.98 +/- 2.17 D using a multipass multizone technique (MP, 3 zones); and 25 eyes were corrected for myopia of -5.03 +/- 1.98 D using new software which enables the creation of three ablation zones in a single-pass fashion (MZ). RESULTS: Mean refractive error (+/-SD) at the first post-operative month was +1.34 +/- 1.00 D for SA, +1.78 +/- 1.49 D for MP, and +0.90 +/- 0.75 D for MZ. Eyes treated by MZ had significantly better uncorrected visual acuity than those treated by SA (P=0.04). Corneal topography revealed a central island in 14 eyes (56%) of those treated by SA and 2 eyes (8%) of those treated by MZ, but in no eyes treated by MP. Six months after treatment, mean refractive error was +0.28 +/- 0.75 D for SA, +0.36 +/- 0.92 D for MP, and - 0.07 +/- 0.77 D for MZ. CONCLUSION: MZ ablation induced less hyperopic shift at 1 month, thus leading to a faster visual recovery and better uncorrected visual acuity. MP and MZ ablations proved effective in preventing central island formation, thus inducing less visual disturbances than SA ablations. These results suggest that MZ ablation may be preferable to the other techniques in performing myopic PRK. PMID- 8724153 TI - Multizone versus monozone in the treatment of high and moderate myopia with an excimer laser. AB - BACKGROUND AND OBJECTIVE: Numerous modifications have been proposed to photorefractive keratectomy (PRK) in the treatment of myopia in excess of 6.00 diopters (D). One of the techniques employed in an attempt to achieve better refractive results and to avoid, above all, the appearance of an intense haze, is the technique known as multizone. PATIENTS AND METHODS: We have attempted, in a prospective study, to compare the refractive results and the complications of monozone and multizone techniques for the treatment of moderate and high myopia. We performed 139 monozone PRKs and 30 multizone PRKs with a one-year follow-up. RESULTS: We did not find statistically significant differences between the two groups either in the degree of correction obtained nor in the final visual acuity. A statistically significant difference was found only in the number of cases with severe haze (greater than 3) which proved more frequent in the monozone group. CONCLUSION: Our results suggest the techniques are comparable in predictability and effectiveness. However, the incidence of severe haze was lower in the multizone group, which may render its use preferable in the treatment of myopias in excess of 6.00 D. PMID- 8724154 TI - Topographic comparison of monozone, multipass and multizone ablations for myopic photorefractive keratectomy. AB - BACKGROUND AND OBJECTIVE: Multizone and multipass ablation techniques have been developed in an attempt to improve results over monozone photorefractive keratectomy (PRK). This study was conducted to evaluate the results obtained in the treatment of myopia with multizone PRK in comparison with monozone ablation in low myopia, and with multipass ablation in high myopia. PATIENTS AND METHODS: Sixty-two eyes with low myopia (-1.90 to -6.25 diopters [D]) were divided into two groups: 42 eyes were treated with a 5.0 mm monozone ablation and compared with 20 eyes treated with a multizone software using 5.0-5.7-6.5 mm optical ones and a 100-60-40% algorithm. Thirty-four eyes with high myopia (-6.50 to -12.00 D) were also divided into two groups: 17 eyes were treated with a multipass technique dividing the attempted correction in three successive treatments with 5.0-6.0-6.5 mm optical zones and 50-30-20% of the total correction, and compared with 17 eyes treated with the multizone software using the same parameters as in the low myopia group. RESULTS: Multizone treatment induced less initial hyperopic shift than monozone or multipass techniques, with significant differences at 15 days and 1 month. At 6 months after surgery 100% of low myopes and 88.24% of high myopes treated with the multizone software were within +/- 1.00 D of emmetropia as compared with 90.5% of the monozone eyes and 52.9% of the multipass patients. CONCLUSION: In low myopes this improvement seems to be related only to the increase in ablation diameter, but in high myopes we found a great improvement in the ablation profile and a shortening of the procedure as the likely factors of the better results obtained with multizone ablation. PMID- 8724155 TI - The treatment of pain following excimer laser photorefractive keratectomy: additive effect of local anesthetic drops, topical diclofenac, and bandage soft contact. AB - BACKGROUND AND OBJECTIVE: Post-operative pain is a transient but undesirable side effect of photorefractive keratectomy (PRK). Pain control after excimer laser PRK was assessed in 112 eyes that were divided into 6 pain management groups. PATIENTS AND METHODS: "Pain at its worst" was measured using a visual analog scale. The groups are arranged in order, Group 6 having experienced the most effective relief of pain and Group 1 the least relief of pain: 1) Topical amethocaine/tetracaine, 2) Topical amethocaine/tetracaine plus bandage contact lens, 3) Topical diclofenac, 4) Topical diclofenac plus bandage contact lens, 5) Topical amethocaine/tetracaine plus diclofenac, and 6) Topical amethocaine/tetracaine, diclofenac plus bandage contact lens. All patients were given 10 tablets of co-dydramol (10 mg dihydrocodeine and 500 mg paracetamol per tablet). They were to be used every 4 hours, but patients were specifically instructed not to take the tablets unless the topical pain management was inadequate for their needs. RESULTS AND CONCLUSION: Group 6 was dramatically better than any other group. In fact, 31% of Group 6 patients had no pain whatsoever. Removing any one of the 3 pain management tools resulted in much less pain control. Day 1 pain was also much more effectively controlled by the Group 6 management compared with those of the other groups. Group 6 patients took fewer co-dydramol tablets than any of the other groups. In addition, Group 6 scored best n several other areas that were studied: a) Day 1 light sensitivity; b) Day 1 tearing; c) Day 1 drowsiness; and d) number of hours slept on the first night. The epithelium healed in all patients but one Group 5 and one Group 6 patient had an epithelial defect for longer than 7 days. There was no instance of infectious keratitis, nor was there an instance of the sterile keratitis that has been associated with the use of topical non-steroidal anti-inflammatory drops. PMID- 8724156 TI - Effect of steroids and nonsteroidal anti-inflammatory agents on stromal wound healing following excimer laser keratectomy in rabbits. AB - BACKGROUND AND OBJECTIVE: The effect of topical corticosteroids and nonsteroidal anti-inflammatory drugs (NSAIDs) on stromal wound healing after excimer laser keratectomy was evaluated quantitatively. MATERIALS AND METHODS: Excimer laser keratectomy was performed in each eye of 75 rabbits and the animals were divided into five groups according to planned post-operative medications: group I, 0.1% dexamethasone; group II, 0.1% fluorometholone; group III, 0.03% flurbiprofen; group IV, 0.1% dexamethasone and 0.03% flurbiprofen; and control group V, balanced salt solution. All treatment regimens were instilled four times daily. After dichlorotriazinyl aminofluorescein (DTAF) or hematoxylin-eosin staining, the thickness of new collagen and keratocyte density were quantified by image analysis system. In addition, the ultrastructural alterations in stroma were examined by transmission electron microscopy. RESULTS: At four weeks, keratocyte density had peaked and new collagen identified in all groups. At 12 weeks, new collagen thickness and keratocyte density of the control were significantly different from that of each experimental group (P < 0.05). Transmission electron microscopy showed the activated keratocytes and extracellular matrixes deposition. At 12 weeks, the structural changes had much subsided in each experimental group but sustained in control. CONCLUSION: Topically applied corticosteroids and NSAIDs can modulate stromal wound healing after excimer laser keratectomy and the effect of dexamethasone, which can be evaluated by the quantitative analysis of the morphologic changes, is the most prominent. PMID- 8724157 TI - Removal of epithelium and scraping the underlying stroma as treatment for photorefractive keratectomy overcorrection or undercorrection of myopia. AB - BACKGROUND AND OBJECTIVE: A minority of patients who undergo excimer laser photorefractive keratectomy are overcorrected to the extent that they are symptomatic; they tend to be "poor healers" with little post-excimer subepithelial haze and a thinned epithelial layer. The reverse situation can also occur where "aggressive healing" results in undercorrection, excessive haze and a thickened epithelial layer. PATIENTS AND METHODS: Overcorrection was treated in 19 eyes by removal of the corneal epithelium overlying the treated cornea with a disposable number 15 scalpel blade under topical anesthesia at the slit lamp. The exposed stroma was gently abraded with the blade in order to create inflammation and the laying down of tissue to partially resteepen the cornea. Eight eyes with undercorrection had "hyperplastic epithelium" removed with the number 15 blade. The intention was to abrade underlying subepithelial haze only if it was felt that a plaque of "haze" could be removed by so doing; in fact, no plaques were removed and it was difficult to remove the epithelium without associated trauma to the superficial stroma. RESULTS: The mean refraction of the overcorrected eyes was +2.77 diopters (D) compared with +0.68 D at 12 months post-scraping. This can be contrasted with -3.11 D prior to scraping, and -4.67 D at 12 months for the undercorrected eyes. The only difference that could be detected between the overcorrected and undercorrected eyes was that the mean pre-excimer refraction of the overcorrected patients was -5.68 D, whereas that of the undercorrected patients was -8.22 D. It was not possible to predict which overcorrected eyes would do well after scraping. CONCLUSION: Scraping is a very effective treatment of overcorrection after photorefractive keratectomy. Scraping as a treatment of undercorrection should not be undertaken. PMID- 8724158 TI - Treatment of astigmatism associated with myopia or hyperopia with the holmium laser: second year follow-up. AB - BACKGROUND AND OBJECTIVE: In 1993, the Summit Technology apogee system for treating astigmatism was not available, so (to avoid the deep cuts of astigmatic keratotomy) the Summit Holmium: YAG laser was used to treat astigmatism associated with myopia; an excimer laser photorefractive keratectomy was to be performed when the keratometry readings had stabilized. The holmium:YAG laser also was utilized to concurrently treat hyperopia and astigmatism. PATIENTS AND METHODS: For myopic astigmatism, four holmium spots were administered, two on either side of the optical zone. An optical zone of 6.5 mm was used for keratometric astigmatism of 4.00 diopters (D) or greater; 7.0 mm for 3.00 to 3.90 D; 7.5 mm for 1.50 to 2.90 D. For hyperopic astigmatism, two extra spots were placed in the inner treatment ring, one on either side of the flattest meridian for 1.50 to 2.90 D of keratometric astigmatism. By omitting the outer ring treatments in the steeper meridian and using four extra spots, two on either side of the flattest meridian in the inner ring, 3.00 D or more was treated. RESULTS: Thirty-one myopic eyes were treated; five were lost to follow-up; 12 have subsequently had a photoastigmatic refractive keratectomy (PARK); six have had a PRK; eight had had no further procedures. Seven eyes now have 18 months of follow up or more (and have not had a PARK); the four successes are presented in detail and it appears that their keratometry readings are nearly stable. CONCLUSION: Four of 7 (57%) myopic eyes, which had holmium laser treatment of associated astigmatism, can be considered as successes after 18 months or more follow-up. Keratometry readings are nearly stable; further follow-up is required to determine if regression continues. The treatment of astigmatism simultaneously with hyperopia treatment with the holmium laser was not successful. PMID- 8724159 TI - One year follow-up of photoastigmatic keratectomy (PARK) with the VISX 20/20 excimer laser. AB - BACKGROUND AND OBJECTIVE: One hundred thirteen eyes were treated with photoastigmatic keratectomy using the VISX 20/20 excimer laser. The results were evaluated after 1 year. PATIENTS AND METHODS: The patients were divided into 2 groups, low astigmatism group with less than 2.00 diopters (D) of pre-operative astigmatism, the second group having 2.00 D or more. RESULTS: Astigmatism was reduced by 43.9% in the low astigmatic group and by 71.7% in the high astigmatic group. Eighty percent of the eyes in the low astigmatic group and 90% in the high astigmatic group achieved an uncorrected visual acuity of 20/40 (0.5) or better. Eighty-four percent in the low astigmatic group and 80% in the high astigmatic group were within +/- 1.00 D in spherical equivalent results. The mean spherical equivalent results were about -0.45 D in both groups using this algorithm. CONCLUSION: The results are very promising at this early stage of algorithm development. PMID- 8724160 TI - Phototherapeutic keratectomy for corneal scars. AB - BACKGROUND AND OBJECTIVE: Phototherapeutic keratectomy (PTK) is an effective method of treating a variety of corneal disorders. We report our experience in using PRK to treat corneal scars. PATIENTS AND METHODS: We retrospectively analyzed the data from a cohort of 22 eyes with corneal scars. The etiology of the scar was recorded for each patient. The change in best corrected visual acuity, spherical equivalent, and astigmatism were evaluated at one and three months after surgery. RESULTS: At three months, seven of 18 eyes (39%) had improved 2 or more lines of best corrected visual acuity and 3 eyes (16%) had lost 2 or more lines. Eleven of 18 eyes (61%) had a hyperopic shift of more than 1.00 diopter (D) and eight of 18 eyes (44%) had a hyperopic shift of more than 2.00 D. There was a mean reduction in astigmatism of 0.67 +/- 4.00 D. Four of eight eyes with traumatic scars had significant improvement as compared to none of five eyes with infectious scars. CONCLUSION: PTK is a relatively safe and effective means of treating corneal scars and thereby may offer an alternative to corneal transplantation. PMID- 8724161 TI - Laser assisted in situ keratomileusis for high myopia. AB - BACKGROUND AND OBJECTIVE: We present the clinical results of laser assisted in situ keratomileusis (LASIK) to correct myopia of more than -10.00 diopters (D). PATIENTS AND METHODS: Eighteen eyes are included in this study. Pre-operative spherical equivalent was -16.40 D, range from -10.50 to -21.50 D. Hinge corneal cap techniques were used with automated microkeratome (Steinway, USA). The thickness of the flap was 130 microns. The stromal bed was ablated with the OmniMed II excimer laser, using the standard MKM program (Summit Technology, Inc., Waltham, MA). The ablation zone was between 4.2 and 5.0 mm. RESULTS: At 6 months postoperatively, the mean spherical equivalent was +0.14 D with a range of +3.50 to -2.00 D. A result within 2.00 D of the desired correction was achieved in 93% of eyes, while a result within 1.00 D was achieved in 47%. Uncorrected visual acuity was improved in all cases. One eye lost greater than two lines of pre-operative best corrected vision due to macular hemorrhage. CONCLUSION: These data show that the LASIK procedure appears to be effective for correcting high myopia. PMID- 8724162 TI - The Martines enhancement technique for correcting residual myopia following laser assisted in situ keratomileusis. AB - BACKGROUND AND OBJECTIVE: Laser assisted in situ keratomileusis (LASIK) is highly effective for correcting moderate to severe levels of myopia with rapid visual recovery, minimal pain and easier post-operative management compared with Photorefractive keratectomy. Nevertheless, undercorrection may occur. The purpose of this paper is to evaluate 14 undercorrected LASIK cases in which the cap was dissected and the stromal bed re-ablated using a method developed by one of the authors (EM). PATIENTS AND METHODS: Among a series of 297 LASIK procedures, visual and refractive results of 14 cases undergoing enhancement were evaluated. All cases were targeted for plano refraction. RESULTS: All 14 cases were followed at least 6 months and 7 (50%) have been followed two years. Mean pre-LASIK myopia of -14.60 +/- 4.90 D had been reduced to a mean of -4.50 +/- 2.60 D prior to enhancement. Following enhancement mean spherical equivalent was -0.22 D at 6 months. Results were stable at two years. All cases were within 1.25 D of target and 11 of 14 were within one diopter. Uncorrected vision was improved in all cases. Best corrected vision was maintained or improved in all but one case. This case had a decentered ablation. CONCLUSION: Laser enhancement of undercorrected LASIK cases using the Martines Technique was predictable and efficacious. PMID- 8724163 TI - LASIK for high myopia: one year experience. AB - BACKGROUND AND OBJECTIVE: Laser in situ keratomileusis (LASIK) is a technique combining the advantages of lamellar corneal surgery (not disturbing Bowman's layer) with the precision of excimer laser surgery. It can be used to correct myopia in a wide range of diopters. In this study we present our data on the first 34 consecutive cases of myopia correction by LASIK. PATIENTS AND METHODS: Thirty four eyes of 27 patients with myopia ranging from -10.00 to -22.50 diopters (D) were operated with LASIK. The follow-up was from six months to one year. The mean age was 33.02 +/- 7.98 years, 32.4% were males and 67.6% were females. Surgery was always performed with the Chiron Automated corneal shaper resecting a corneal flap of 160 mu and then ablating the stromal bed (OZ 4.50 mm) with the Summit OmniMed 5.0 excimer laser. The parameters used to assess the results were predictability, safety (variation of spectacle corrected visual acuity), stability, and patient satisfaction. RESULTS: Of the patients, 67.65% were between -1.00 and +1.00 at 6 months; 44.11% of eyes retained the same spectacle corrected visual acuity, 11.76% gained 1 line, 26.47% gained 2 lines, 8.82% lost 1 line and 8.82% lost 2 lines. Refraction at 1 month was 0.07 +/- 1.99 (range, -4.00 to +4.00); at 3 months 0.61 +/- 1.69 (range, -4.25 to +2.25); and at 6 months 0.81 +/- 1.71 (range, -4.75 to +2.25). Of the patients, 85.3% were happy with the procedure. Complications included two dislocated caps in the immediately post-operative period and two cases of epithelial ingrowth under the cap. Two eyes developed irregular astigmatism. These complications were found in the first two sessions of surgery and are related to the learning curve of the method. CONCLUSION: Results suggest that LASIK is a viable method for correction of high myopia with few complications and may have advantages over keratomileusis and PRK as it combines the best features of both methods--it preserves Bowman's layer and has the excimer laser precision. Refining the nomogram of ablation should result in even better results. PMID- 8724164 TI - Holmium:YAG laser thermokeratoplasty to correct hyperopia: two years follow-up. AB - BACKGROUND AND OBJECTIVE: Treatment of hyperopia is more of a challenge than PRK but the development of the holmium:YAG laser has provided a more controlled way of carrying out laser thermokeratoplasty (LTK). PATIENTS AND METHODS: Twenty two eyes with hyperopia were treated with a Summit Technology OmniMed holmium laser by placing two rings of eight laser spots at 6.5 and 9 mm (centred on the visual axis) to produce a 4.00 diopters (D) correction. RESULTS: An average +2.10 D refractive correction was achieved in the 17 eyes with no induced astigmatism with an accuracy of 25% within 1.00 D, 60% within 2.00 D and 100% within 3.00 D. However, significant astigmatism (+1.25 to +2.5 Dcyl) was produced in 23% of eyes from 6 months post-laser and these required astigmatic LTK correction. CONCLUSIONS: The overall results were disappointing in that there was an approximate 50% regression at two years post-LTK. However, the rate of regression was very slow at 24 months, and most patients remarked how well they could see in the first few months following the treatment. PMID- 8724165 TI - Functional electrical stimulation systems: what have we accomplished, where are we going? PMID- 8724166 TI - Functional neuromuscular stimulation (FNS). PMID- 8724167 TI - Challenges to clinical deployment of upper limb neuroprostheses. AB - The technology for functional neuromuscular stimulation (FNS) as a means of providing upper limb function to people with tetraplegia has been under development by three clinical research groups for almost two decades. This paper presents the current status of the clinical trials of three FNS systems: a noninvasive system built into a cosmetic forearm splint, a 30-channel percutaneous system, and an 8-channel implantable system. The complexity of FNS systems and the unique characteristics of the individuals to whom they are applied combine to create many clinical and technical challenges that must be addressed before the devices can be deployed. The emerging challenges to widespread clinical introduction of FNS systems for hand and arm function are identified and analyzed. In addition to the demands of designed and conducting the clinical trials to satisfy regulatory requirements, the lack of knowledge, skepticism, and the complacency on the part of potential FNS recipients, as well as of rehabilitation professionals, must be overcome through education and careful consideration of economic and societal factors in the design of clinical systems. PMID- 8724168 TI - Electrical stimulation to restore respiration. AB - Electrical stimulation has been used for over 25 years to restore breathing to patients with high quadriplegia causing respiratory paralysis and patients with central alveolar hypoventilation. Three groups have developed electrical pacing systems for long-term support of respiration in humans. These systems consist of electrodes implanted on the phrenic nerves, connected by leads to a stimulator implanted under the skin, and powered and controlled from a battery-powered transmitter outside the body. The systems differ principally in the electrode design and stimulation waveform. Approximately 1,000 people worldwide have received one of the three phrenic pacing devices, most with strongly positive results: reduced risk of tracheal problems and chronic infection, the ability to speak and smell more normally, reduced risk of accidental interruption of respiration, greater independence, and reduced costs and time for ventilatory care. For patients with partial lesions of the phrenic nerves, intercostal muscle stimulation may supplement respiration. PMID- 8724169 TI - Dynamic myoplasty: surgical transfer and stimulation of skeletal muscle for functional substitution or enhancement. AB - Dynamic myoplasty combines muscle transfer with electrical stimulation to provide contractile function that augments or replaces impaired organ function. Dynamic cardiomyoplasty was the first clinical application in which a skeletal muscle, latissimus dorsi, was transferred and stimulated to provide cardiac assistance, a function different from its original one. The problem of early muscle fatigue that was encountered in the initial implementation of the method was solved by training the muscle with electrical stimulation and thus changing its fiber composition. With intramuscular electrodes, the conditioned latissimus dorsi is stimulated in synchrony with the heart muscle. Safeguards are built into the two channel implanted stimulator to avoid excessively high pulse rates. Clinicians report that 80% of patients with moderate to severe heart failure prior to operation showed a clinical improvement of 1.6 New York Heart Association classes. Alternative methods of providing cardiac assistance that are also being investigated include wrapping the muscle around the aorta, creating a skeletal muscle ventricle, and using the muscle to power an implantable pump. These latter techniques are still under preclinical investigation. Compared with heart transplant, cardiomyoplasty has the great advantage of not being subject to tissue rejection. The second principal application of dynamic myoplasty is treatment of fecal incontinence through creation of an electrically stimulated skeletal muscle neosphincter (ESMNS). The gracilis muscle of the leg is mobilized, wrapped around the anal canal, and conditioned with electrical stimulation to become more fatigue resistant. To achieve continence, the muscle is continuously stimulated except when the patient wishes to defecate. Overall success rates in achieving continence are 60-65%. Both cardiomyoplasty and the ESMNS technique, and their associated devices, are being refined through ongoing clinical trials. PMID- 8724171 TI - New control strategies for neuroprosthetic systems. AB - The availability of techniques to artificially excite paralyzed muscles opens enormous potential for restoring both upper and lower extremity movements with neuroprostheses. Neuroprostheses must stimulate muscle, and control and regulate the artificial movements produced. Control methods to accomplish these tasks include feedforward (open-loop), feedback, and adaptive control. Feedforward control requires a great deal of information about the biomechanical behavior of the limb. For the upper extremity, an artificial motor program was developed to provide such movement program input to a neuroprosthesis. In lower extremity control, one group achieved their best results by attempting to meet naturally perceived gait objectives rather than to follow an exact joint angle trajectory. Adaptive feedforward control, as implemented in the cycle-to-cycle controller, gave good compensation for the gradual decrease in performance observed with open loop control. A neural network controller was able to control its system to customize stimulation parameters in order to generate a desired output trajectory in a given individual and to maintain tracking performance in the presence of muscle fatigue. The authors believe that practical FNS control systems must exhibit many of these features of neurophysiological systems. PMID- 8724170 TI - Neural signals for command control and feedback in functional neuromuscular stimulation: a review. AB - In current functional neuromuscular stimulation systems (FNS), control and feedback signals are usually provided by external sensors and switches, which pose problems such as donning and calibration time, cosmesis, and mechanical vulnerability. Artificial sensors are difficult to build and are insufficiently biocompatible and reliable for implantation. With the advent of methods for electrical interfacing with nerves and muscles, natural sensors are being considered as an alternative source of feedback and command signals for FNS. Decision making methods for higher level control can perform equally well with natural or artificial sensors. Recording nerve cuff electrodes have been developed and tested in animals and demonstrated to be feasible in humans for control of dorsiflexion in foot-drop and grasp in quadriplegia. Electromyographic signals, being one thousand times larger than electroneurograms, are easier to measure but have not been able to provide reliable indicators (e.g., of muscle fatigue) that would be useful in FNS systems. Animal studies have shown that information about the shape and movement of arm trajectories can be extracted from brain cortical activity, suggesting that FNS may ultimately be directly controllable from the central nervous system. PMID- 8724172 TI - Technology transfer of neuroprosthetic devices. AB - Despite long development periods for neuroprosthetic devices, the numbers in clinical use or clinical trials are rising, with an estimated 3,000 systems in use today. As they gain experience with the regulatory approval process, developers are learning to conduct research to best prepare for transfer of technology to industry. The track record of the first motor prosthesis to be approved by the United States Food and Drug Administration contains important lessons for a company planning to undergo the regulatory process. Throughout the development of a neuroprosthesis, the capabilities and preferences of the customers who will use it (physicians, surgeons, therapists, and end-users) should be sought out and used in device design. When a device has reached clinical application, particular attention is needed to maximize both the population who will use it and each individual's degree of use (optimal, partial, reluctant). Identification of person-technology mismatches can help to select training strategies and other interventions that can be applied to ensure a good rehabilitation outcome. PMID- 8724173 TI - Social modulation of sex steroid concentrations in the urine of male cichlid fish Oreochromis mossambicus. AB - The relationship between urinary concentrations (free + sulfates + glucuronides) of the steroids testosterone (T), 11-ketotestosterone (11KT), 17 alpha, 20 beta dihydroxy-4-pregnen-3-one (17, 20 beta-P) and 17 alpha, 20 alpha-dihydroxy-4 pregnen-3-one (17, 20 alpha-P), and the social behavior of males of the cichlid fish Oreochromis mossambicus was investigated. After 8 days of isolation none of the steroids were good predictors of social dominance developed after subsequent formation of all-male groups. One day after group formation dominance indexes were good predictors of the urine concentrations of all sex steroids. Dominance indexes and androgen concentrations measured after all-male group formation were positively correlated with territoriality, courtship rate, and nest size. Similar relationships were found for progestins with the exception that they were not correlated with courtship rate. All-male group formation was also accompanied by an increase in urinary sex steroid concentrations in fish that became territorial and a decrease in non-territorial fish with the exception of T, which increased in both groups. Addition of ovulating females caused steroid concentrations to return to levels near isolation, except for 17, 20 alpha-P in territorials, which underwent a large increase. Thus, social interactions may have an important modulatory effect on sex steroid concentrations in O. mossambicus. PMID- 8724174 TI - Dominance, nutritional state, and growth hormone levels in rainbow trout (Oncorhynchus mykiss). AB - This study addressed three questions concerning interactions between physiology and dominance in juvenile rainbow trout: (1) the validity of a model predicting a time-dependent effect of fasting on competitive ability (i.e., the ability to obtain contested food items) was tested in a series of dominance trials between fed and progressively more fasted trout, as was (2) the association between fasting and plasma growth hormone levels. (3) The relationship between plasma growth hormone levels and the competitive ability of individual trout was also studied. The main results were as follows: (1) The predictions of the time dependent model were supported by the fasting-dominance experiment. After 3 days, fasted fish were dominant over fed fish, whereas after 6 and 9 days, the competitive ability of fed and fasted fish was similar. After 12 days, there was a tendency for fed fish to be dominant over their fasted competitors. (2) Sampling of plasma from fed and fasted trout, after 3, 6, 9, and 12 days, demonstrated that plasma growth hormone levels increases in food-deprived rainbow trout after more than 6 days of fasting, which is consistent with previous work. (3) No difference in plasma growth hormone levels was found between paired dominant and subordinate trout. Possible interactions between nutritional state, growth hormone levels, and dominance, and their implications are discussed. PMID- 8724175 TI - Progesterone does not inhibit aggression induced by testosterone metabolites in castrated male mice. AB - Sufficient data exist indicating that testosterone (T) or metabolites of T, i.e., dihydrotestosterone (DHT) and estrogen (E), induce aggressive behavior. The effects of T have also been shown to be suppressed by concurrent administration of progesterone (P). It is unknown if the effects of P are exerted on T directly or upon the T metabolites. The objective of this experiment was to evaluate the inhibitory effects of P on aggression induced by T and T metabolites. Seven male mice were randomly allocated to one of the following treatment groups: castration (C), C + T, C + 5 alpha-dihydrotestosterone (DHT), C + estrogen (E), C + DHT + E (DE), T + P, and DE + P. The males were then placed with androgenized females in three 10-min trials to determine the effects of each treatment on hormone induced aggression. The degree of aggressive behavior displayed was measured by number of trials where attacks occurred (N), mean latency to attack (L), frequency of attacks (AF), duration of attacks (D), and a composite index score (INDEX). All androgen treated groups showed significantly higher degrees of aggression compared to C and T + P animals. P inhibited aggression in T-treated males but did not inhibit aggression in the DE treated group. The data suggest that P inhibits the aggressive effects of T by acting on receptors of the metabolite producing enzymes or by direct competitive binding for 5 alpha-reductase. PMID- 8724176 TI - Odors providing sexual information in Djungarian hamsters: evidence for an across odor code. AB - Experiments were conducted to determine the repertoire of odors that are involved in sexual recognition and communication about reproductive state in Djungarian hamsters (Phodopus campbelli). By examining the preferences of males for odors of females over those of males, it was found that some odors support preferences across female reproductive states, other odors induce preferences only during estrus, and yet other odors do not elicit sexual preferences. Similarly, female preferences for male odors was restricted to just some scents. Males were also tested for their preferences for female odors from different reproductive states (diestrous, day before parturition, post-partum estrus). Four different odors were found to vary in attractiveness with these reproductive states, and there were three different patterns of change: (1) mouth and urine odors were attractive just during post-partum estrous; (2) midventral gland was most attractive the day before parturition; (3) vaginal secretions showed a graded pattern of attractiveness, peaking at post-partum estrus. These results show that different odors provide some redundant and some different information, and they suggest that an across-odor code could be used to provide very precise information about a female's reproductive state. PMID- 8724177 TI - Allopregnanolone (THP) mediates anesthetic effects of progesterone in rat brain. AB - Intravenous infusion of the progesterone or that of progesterone 5 alpha-reduced metabolite, 3 alpha-hydroxy-5 alpha-pregnan-20-one (THP), induces the loss of righting reflex in freely moving rats at the doses of 49 +/- 15 mg/kg or 5.6 +/- 2.2 mg/kg, respectively. The recovery time of righting reflex was 71 +/- 12 min and 21 +/- 5 min for progesterone and THP, respectively. The time course of brain concentrations of THP, but not that of progesterone, correlated with the loss and the recovery of righting reflex. The pretreatment of animals with uncompetitive inhibitor of 5 alpha-reductase 17 beta (N,N-diisopropylcarbamoyl)-androst-3-diene 3-carboxylic acid (SKF 105111) significantly reduces the anesthetic activity of progesterone, but not that of THP. Following progesterone infusion brain level of THP in SKF 105111 pretreated rats was 12% that of vehicle-treated control, and the level of progesterone was 160%. No effect of SKF 105111 on brain THP level was detected in animals infused with THP. These results demonstrate that anesthetic effect of progesterone is mediated through its conversion to THP and support the hypothesis that endogenous metabolites of progesterone may be involved in the regulation of behavior in rats. PMID- 8724178 TI - The hormonal induction of mating behavior in female gray short-tailed opossums (Monodelphis domestica). AB - The activating effects of estradiol alone and of estradiol plus progesterone on copulation and related behavior were examined in ovariectomized female gray short tailed opossums. Following receipt of subcutaneous estradiol (E) or cholesterol (C) implants, E-females showed less screeching threat behavior toward intact males than C-females. Intact males showed more precopulatory behavior toward E- than toward C-females, and 20% of E-females and no C-females copulated with males. In a second experiment, females that had previously received subcutaneous E implants received injections of either corn oil alone (E + OIL) or 500 micrograms progesterone in corn oil (E + P) 4 hr before behavioral testing. Of E + P-females, 88% copulated with males, compared to 38% of E + OIL-females. All females that copulated in both experiments showed anogenital dragging just prior to copulation. Hormonal induction of estrous behavior and copulation has not been reported previously in a female marsupial. PMID- 8724180 TI - Levels of plasma corticosterone and testosterone in male copperheads (Agkistrodon contortrix) following staged fights. AB - Fighting behavior between male copperheads (Agkistrodon contortrix) occurs during the two mating periods (late summer/fall and spring) to gain priority of access to females. Fights are characterized by prominent vertical challenge displays, swaying, and a high degree of physical contact that does not involve biting. At the moment of subordination, losers retreat quickly from fights and winners respond by chasing. Subsequently, losers do not participate in further challenge displays or fighting for at least 7 days, and also they show behavioral signs of stress, which includes submissive acts and suppression of sexual behavior. The goal of this study was to determine whether or not losers show elevated levels of plasma corticosterone (B) and depressed levels of plasma testosterone (T) relative to winners and controls. Winners and losers were produced in 13 staged trials. Two different controls (N = 26) were run. Males with no recent agonistic experience were (1) tested in the fighting arena in the absence of a competitive male but paired with a single female (N = 13), and (2) tested alone in their cages (N = 13). All trials, including controls, were conducted in spring and late summer. Mean B in losers at 1-hr postfight was significantly greater than in winners and both control groups in both seasons. Mean T was significantly greater in late summer in all groups, as expected, but in each season was not significantly different between the groups. Levels of B and T were not correlated with SVL, mass, or duration of fighting. This study provides further support for the social insensitivity/challenge hypotheses and is the first to document postfight B and T levels in snakes. PMID- 8724179 TI - Corticotropin-releasing factor enhances locomotion and medullary neuronal firing in an amphibian. AB - Corticotropin-releasing factor (CRF) administration has been shown to act centrally to enhance locomotion in rats and amphibians. In the present study we used an amphibian, the roughskin newt (Taricha granulosa), to characterize changes in medullary neuronal activity associated with CRF-induced walking and swimming in animals chronically implanted with fine-wire microelectrodes. Neuronal activity was recorded from the raphe and adjacent reticular region of the rostral medulla. Under baseline conditions most of the recorded neurons showed low to moderate amounts of neuronal activity during periods of immobility and pronounced increases in firing that were time-locked with episodes of walking. These neurons sometimes showed further increases in discharge during swimming. Injections of CRF but not saline into the lateral ventricle produced a rapidly appearing increase in walking and pronounced changes (mostly increases) in firing rates of the medullary neurons. CRF produced diverse changes in patterns of firing in different neurons, but for these neurons as a group, the effects of CRF showed a close temporal association with the onset and expression of the peptide's effect on locomotion. In neurons that were active exclusively during movement prior to CRF treatment, the post-CRF increase in firing was evident during episodes of walking; in other neurons that also were spontaneously active during immobility prior to CRF infusion, post-CRF activity changes were evident during immobility as well as during episodes of locomotion. Thus, a principal effect of CRF was to potentiate the level of neuronal firing in a population of medullary neurons with locomotor-related properties. Due to the route of administration CRF may have acted on multiple central nervous system sites to enhance locomotion, but the results are consistent with neurophysiological effects involving medullary locomotion-regulating neurons. PMID- 8724181 TI - Corticosterone and interchromosomal race discrimination in the house mouse. AB - We report results of a study on emotional arousal, as reflected by corticosterone variation, following dyadic encounters within and between chromosomal races of the house mouse (Mus musculus domesticus). Three contiguous chromosomal races from northeast Scotland were analyzed (2n = 32, 2n = 36, 2n = 40). Within-race encounters resulted in similar average corticosterone levels in mice from the three races. For all three races, certain of the between-race encounters resulted in a significant increase in corticosterone relative to those following within race encounters. These results confirm our previous observations of a differential behavioral treatment of house mice from the same vs from a different chromosomal race, and support our proposition for behavioral discrimination between chromosomal races of this species in northeast Scotland. PMID- 8724182 TI - Effects of testosterone and 7 alpha-methyl-19-nortestosterone (MENT) on sexual and aggressive behaviors in two inbred strains of male mice. AB - Behavioral and endocrine effects of a synthetic androgen, 7 alpha-methyl-19 nortestosterone (MENT), which is not 5 alpha-reduced to dihydrotestosterone, were compared to those of testosterone in two inbred strains of male mice, C57BL/6J and DBA/2J, in two experiments. In the first experiment, seminal vesicle (SV) weights, kidney weights, and circulating steroid levels were examined in castrated mice treated with three doses of testosterone (3.125, 12.5, or 50 micrograms/day) or four doses of MENT (1, 4, 16, or 64 micrograms/day) for 2 weeks to determine the optimal replacement levels of the two androgens for behavioral studies. Both testosterone and MENT dose-dependently increased the SV weights that were greatly reduced, in both strains, by castration. MENT was more effective than testosterone in increasing SV weights, fully restoring them to intact levels in both strains, at the dose of 4 micrograms/day. At the dose of 12.5 micrograms/day, testosterone restored the SV weights completely in C57BL/6J and up to 80% in DBA/2J mice. DBA/2J mice were more sensitive than C57BL/6J mice to both androgens, as measured by kidney weights, although circulating levels of either steroid were very similar between the two strains of mice. In the second experiment, we investigated the effects of testosterone (12.5 micrograms/day) and MENT (4 micrograms/day) on sexual and aggressive behaviors. In each strain, MENT treated and testosterone-treated mice showed similar numbers of mounts or intromissions. MENT was equally effective as testosterone to fully (C57BL/6J) or partially (DBA/2J) restore sexual behaviors as well as the SV weights to the intact levels. In contrast, MENT-treated mice of both strains were much less aggressive than testosterone-treated mice. In both C57BL/6J and DBA/2J mice, testosterone fully restored aggression to the intact levels as measured by aggression latency, number of aggressive bouts, and duration of aggression, whereas aggressive behaviors of the MENT-treated groups were not different from those of the castrated control groups. These results suggest that MENT can restore both male sexual behaviors and reproductive organ weights as effectively as testosterone, at one-third of the testosterone dose, without stimulating male aggressive behaviors. PMID- 8724183 TI - Behavioral strategy and the physiological stress response in rainbow trout exposed to severe hypoxia. AB - In higher vertebrates, two opposite behavioral coping strategies can be distinguished that are associated by a typical neuroendocrine pattern. Little is known about the individual variation in the stress response in lower vertebrates such as teleosts. In the present study, rainbow trout were fitted with an indwelling aortic catheter for repeated blood sampling and exposed to severe hypoxia and subsequent recovery and their behavior was characterized semiquantitatively during hypoxia. Blood levels of catecholamines, cortisol, glucose, FFA, lactate, and electrolytes were measured. About 60% of the fish survived the experiment whereas the others died during the recovery period. Behavioral strategy appeared to be highly related to survival since nonsurviving fish displayed strenuous avoidance behavior involving burst type activity whereas surviving fish did not panic and remained quiet. These behavioral differences were associated with marked differences in plasma catecholamine levels, which were 4- to 5-fold higher in nonsurviving fish as compared to survivors whereas the cortisol response tends to be lower in nonsurviving fish. Plasma lactate levels in nonsurvivors were 4- to 5-fold higher as compared to survivors while a severe hyperkalemia developed during recovery indicating the loss of intracellular homeostasis. The individual differences in behavioral concepts and neuroendocrine activation observed in rainbow trout during stress show great similarity with the active and passive coping strategies distinguished in higher vertebrates and may be determinant for survival during hypoxia. PMID- 8724184 TI - Long-term effects of androgen treatment on fear reactions in ewes. AB - We have previously demonstrated that testosterone treatment reduces fear reactions of ewes subjected to daily injections of testosterone propionate (10 mg/day) for 56 days. The long-term effects of this treatment were studied in the same ewes and using the same battery of fear-eliciting tests (isolation from conspecifics, surprise, and presence of a human) 4 and 8 months after the injections were terminated. Twenty-six behavioral items related to fear were recorded. A number of differences in the behavior of treated vs control animals indicate that the former are still less fearful, especially in the surprise and human tests. Overall fearfulness scores of treated and control ewes (the higher the score, the more fearful the animals) were 7.3 +/- 1.7 vs 9.7 +/- 1.9, (P < 0.05) for the surprise test, and 7.0 +/- 1.4 vs 10.0 +/- 2.0 (P < 0.01) for the human test at 4 months, and 6.8 +/- 1.4 vs 8.2 +/- 1.1 (P < 0.05) for the human test at 8 months. Thus, not only does testosterone treatment reduce fear reactions in ewes but it also has a long lasting effect on such reactions. PMID- 8724185 TI - Spinal surgery: where is it coming from? what is it? where is it going to? PMID- 8724186 TI - Threshold values for supine and standing Cobb angles and rib hump measurements: prognostic factors for scoliosis. AB - Seven parameters recorded at the first clinical examination of 326 growing scoliotic patients were correlated with the speed of progression of the scoliotic curve during a natural history survey period. The parameters were: age; bone age (according to Greulich and Pyle); pubertal and Risser stage; curve shape; rib hump, measured in forward bending in a sitting patient and supine and standing radiographic Cobb angles of the scoliotic curve. The speed of progression of the scoliotic curve was expressed as the annual increase in Cobb angle. It was quantified graphically after plotting the measurements taken from all the radiographic examinations made during the survey. The survey period ranged from 6 months to several years, depending on the rate of progression. It was 6 months only if the scoliotic curve demonstrated worsening of more than 3 degrees at two successive examinations performed at least 3 months apart. The authors aimed to identify the minimum values of curve angle and rib hump, identified at first examination in 95-100% of patients whose parameters at follow-up were above these values (supine angle: 17 degrees; standing angle: 24 degrees; rib hump: 11 mm), therefore demonstrating curve worsening. Then, they analysed how the other parameters such as age, bone age, state of maturation and curve shape influenced these threshold values of rib hump and supine and standing angles. The authors present the threshold values for the whole sample according to the sexual state of maturation and also for each curve shape. They demonstrate that a combination of states of maturation, several measures of the scoliotic curve and curve shape provides the best basis for individual prognosis. PMID- 8724187 TI - The relationship between surface and radiological deformity in adolescent idiopathic scoliosis: effect of change in body position. AB - Three-dimensional (3D) surface deformity of the trunk in adolescent idiopathic scoliosis (AIS) is affected by changes in patient position. Initial quantification of the curve and its associated deformity, as well as subsequent monitoring, relies on both radiological and surface measurements. However, there is often a discrepancy between radiological and apparent surface deformity. The present investigation studied the dynamics of the 3D deformity associated with changes in patient position on 27 patients with AIS. The trunk deformity was quantified in three positions by measuring the angle of thoracic inclination at each vertebral level using a scoliometer. The patients all had full spine radiography in the antero-posterior erect position. Vertebral rotation and lateral spinal curvature were measured from the radiographs. Body position altered the magnitude of the surface deformity over the whole trunk, with the prone position offering the optimum relationship between 3D trunk shape and radiological deformity. This could be attributed to the ease and standardisation of positioning for prone measurements, together with increased patient comfort. It is suggested that adoption of standardised positioning and measurement of surface and radiological deformity will permit consistency of clinical judgement based on these parameters. PMID- 8724188 TI - Vertebral deformities and scoliosis. AB - Scoliosis, especially idiopathic scoliosis, is a complex three-dimensional deformity of the spine in which the vertebral deformities are known, cuneal deformation being the most commonly known deformity but not the only one. We report here data concerning these specific vertebral deformities in chickens. A pinealectomy was performed in a controlled series of animal experiments. This technique induces progressive scoliosis in more than 80% of chickens, with the advantage of being non-aggressive to the spine. Vertebrae included in major thoracolumbar curves were observed in 17 chickens (11 male, 6 female) and classified into three types of vertebral deformities. Vertebral deformity type 1 is characterized by three-dimensional corporeal torsion, which defines the horizontal disorientation of the curve. Vertebral deformities type 2 and 3 define lateral imbalance in the election plane of the curve. Radiological and anatomical data collected throughout the progression of the scoliosis indicate that there is a correlation between structural vertebral deformities and growth/maturation patterns. We compare our results with those reported in literature concerning human idiopathic scoliosis and experimental animal scoliosis. PMID- 8724189 TI - Predominantly posterior instrumentation and fusion in neuromuscular and neurogenic scoliosis in children and adolescents. AB - We studied the results in 46 patients with neuromuscular and neurogenic scoliosis (average age 13.5 years, range 6-19 years) who had had posterior fusion with a modified Luque technique between May 1985 and June 1992. The main criteria to recommend surgery were curve progression, loss of balance when sitting, control of the head and difficulties in wearing an external orthotic support. The mean preoperative curve was 63 degrees, the postoperative value was 24 degrees, representing a correction of about 62%. The average number of stabilized segments was 13. In 39 out of 46 patients, lumbosacral fixation was included in the construct. Failure of implants, pseudarthroses and major losses of correction in purely neuromuscular scolioses could be avoided by using rigid segmental fixation and a dorsolateral fusion with a mixture of autologous and allogenous bone. The scoliosis most difficult to influence was found to be Friedreich's ataxia. In Duchenne muscular dystrophy the best method of treatment was surgery performed as early as possible, i.e. at the time of loss of walking capacity in the case of a scoliosis exceeding 20 degrees and with two consecutive X-rays proving curve progression. Analysis of our series does not confirm the morbidity and complication rates of previous studies. PMID- 8724190 TI - Radiographic evaluation of the upper cervical spine in rheumatoid arthritis: a retrospective analysis. AB - We analysed retrospectively 295 lateral roentgenograms of the cervical spine in 150 patients with classic or definite rheumatoid arthritis. In addition to measuring the atlantodental interval, measurements of the different vertical parameters described by McGregor, Ranawat and Redlund-Johnell and a new measurement method with high reproducibility were described and their results compared statistically. As a control group we analysed 100 lateral roentgenograms of the cervical spine in patients with no inflammatory disease, posttraumatic lesion, tumour or osseous deformity. PMID- 8724191 TI - Short device fixation and early mobilization for burst fractures of the thoracolumbar junction. AB - The authors present a retrospective study based on a homogeneous series of 34 patients with burst fractures of the thoracolumbar junction, fixed using Cotrel Dubousset instrumentation. These patients underwent instrumentation using a short construct of hooks and screws gripping the two vertebrae above the lesion (2HS) and screws and hooks gripping the first vertebra below the lesion (1SH). This construct was therefore called "2HS-1SH". In order to evaluate just the material resistance after getting up, only the patients who were upright on the 4th day without a body cast and with no secondary anterior osteosynthesis were included in this study. Four patients showed some neurological symptoms on admission but recovery was so quick that they could be included in this study. Mean follow-up was 4 years 1 month (range 3 years 1 month and 6 years 2 months). Vertebral and regional kyphosis angles were measured preoperative, postoperatively and at the final follow-up. Functional recovery and complications were analyzed. Mean vertebral kyphosis was 21.2 degrees preoperatively, 3.8 degrees postoperatively and 5.3 degrees at the final follow-up. Regional kyphosis angles were respectively 19.2, 0.2, and 2.7 degrees. We had two cases of deep suppuration, one early and the other late. None of the patients required analgesics for more than 6 months after the operation. Patients returned to work after 5 months on average. The authors concluded that fixation by screw-and-hook constructs is an effective way to stabilize thoracolumbar junction burst fractures. PMID- 8724192 TI - Urinary bladder dysfunction following anterior lumbosacral spine fusion: case report and review of the literature. AB - A 34-year-old woman suffering from chronic degenerative low back pain involving L5-S1 disc space, refractory to conservative treatment, underwent spinal fusion. A combined instrumented posterolateral, followed by anterior, interbody allograft fusion through a left retroperitoneal approach was performed. Postoperatively, the patient was unable to evacuate her bladder and control her micturition. Anal tone and sensation were intact. A self-catheterisation regime was instituted with a diagnosis of parasympathetic nerve injury during the anterior spinal fusion. After a period of 3 months, the patient regained control of urination. We report this case to highlight the importance of protecting the parasympathetic presacral nerve during L5-S1 anterior interbody fusion, as injury to this nerve affects urinary evacuation. PMID- 8724193 TI - Thoracic spinal stenosis in two brothers due to vitamin D-resistant rickets. AB - X-linked hypophosphataemic vitamin D-resistant rickets is a rare cause of spinal canal stenosis. Two brothers with this condition presented in adulthood with thoracic myelopathy due to spinal canal stenosis. Both were treated by laminectomy using diamond-tipped burrs, with symptomatic improvement. PMID- 8724194 TI - Chronic contained rupture of an aortic aneurysm mimicking infective spondylitis. AB - Chronic contained rupture of an abdominal aortic aneurysm is rare. These aneurysms are small and affected patients are usually normotensive. The resultant pseudoaneurysmal sac may cause extensive vertebral erosion. Diagnosis is often delayed as the condition may present with symptoms referable to the lumbar spine. This report is of two cases where the initial diagnosis was infective spondylitis. PMID- 8724195 TI - Vertebral hemangioma presenting with intermittent claudication. AB - The case of a patient with vertebral hemangioma and unusual clinical presentation is reported, with an attempt to explain these unusual clinical complaints. Vertebral hemangioma is a common and often asymptomatic tumor. Neurologic symptoms may appear due to pressure on the neural tissue caused by extraosseous extension. The patient reported here presented with intermittent claudication. Conventional radiography CT, and MRI revealed vertebral hemangioma at T5 and extraosseous extension compressing the spinal cord. Gravity-related vascular dilatation may induce further compression of the spinal cord and, thus, is thought to be the underlying event in the induction of the intermittent clinical symptoms. The lesion was treated with subtotal corpectomy after embolization and fusion with a strut iliac crest graft. At the 9-month follow-up, the patient was without complaints. We conclude that a diagnosis of vertebral hemangioma should be considered in cases of intermittent neurologic symptoms of the lower extremities. PMID- 8724196 TI - Cauda equina syndrome secondary to lumbar spondylodiscitis caused by Streptococcus milleri. AB - A 79-year-old Afro-Caribbean woman presented with a 5-month history of low back pain and a 2-month history of weakness of the lower limbs associated with cauda equina syndrome. A plain X-ray of the lumbar spine showed a collapse of the L4 body consistent with osteomyelitis. An MRI scan of the lumbar spine showed narrowing of the L3/4 disc space, destruction of the adjacent vertebral bodies and an epidural abscess. She was admitted to hospital and treated with a high dose of i.v. antibiotics followed by radical surgical excision of the lesion through a combined anterior and posterior approach with instrumentation. Tissue culture grew Streptococcus milleri. One week after surgery the patient developed septicaemia. A blood culture grew Pseudomonas aeruginosa, which was successfully treated with antibiotics. She eventually recovered bowel and bladder control and regained muscle power in the lower limbs. Streptococcus milleri is a rare causative organism in osteomyelitis, this being only the eighth reported case in the literature. Aggressive surgical treatment combined with a prolonged antibiotic regime is recommended to achieve a satisfactory result. PMID- 8724197 TI - Migration of an acupuncture needle into the medulla oblongata. AB - A case of a delayed lesion of the medulla oblongata caused by migration of an acupuncture needle is presented. The patient was a 60-year-old woman who had undergone embedded-type acupuncture needle treatment around 1975. In 1993 she was admitted to our hospital with a 3-week history of progressive motor and sensory disturbance of her right upper extremity. CT demonstrated that one needle had penetrated the medulla oblongata transversely at the level of the foramen magnum. The needle was removed surgically without any complications. This is the first report of an acupuncture needle migrating in the medulla oblongata. PMID- 8724198 TI - Tuberculosis of the craniocervical junction: two case reports. AB - We report on two children with tuberculosis of the craniocervical junction. Atlantoaxial instability was evident in both patients due to the destruction of the dens and the atlantoaxial ligaments. In addition, one child had tuberculosis involving the anterior arch of the atlas and the anterior rim of the foramen magnum. Despite anti-tuberculosis treatment and immobilization, atlantoaxial instability was evident on flexion/extension views. Successful fusion of C1 and C2 was performed in both patients. PMID- 8724199 TI - Apical hemifacetectomy of the first sacral vertebra in the treatment of an osteoid osteoma. AB - We present the surgical technique of apical hemifacetectomy, which is useful for performing selective resection of lesions located in the upper articular processes avoiding facet joint instability. This surgical technique was used in a 20-year-old patient to remove an osteoid osteoma deeply located in the S1 articular process. PMID- 8724200 TI - Health concerns of artistic women gymnasts. PMID- 8724201 TI - Exercise testing and prescription. Practical recommendations for the sedentary. AB - A sedentary lifestyle is prevalent in most industrialised societies. Persuasive evidence allows us to demonstrate that a physically active lifestyle protects against the development and progression of many chronic diseases. The assessment of sedentary individuals for the purpose of exercise testing and or exercise prescription should always culminate in the determination of the relative risk of the individual for traumatic events which may be precipitated by participation in moderate physical activity. Sedentary individuals may be categorised in a low to high risk stratification as apparently healthy (Class I), higher risk (Class II), or known coronary heart disease and/or symptomatic of chronic disease (Class III). An expanded role for allied health professionals, such as a clinical exercise physiologist, may enhance and extend the services of physicians and nurses as they relate to exercise testing, exercise prescription and preventative healthcare in general. Risk stratification will determine the type of exercise test, the exercise prescription and the exercise environment (low to high levels of supervision). The exercise prescription may include a determination of mode, duration, frequency, intensity, and progression of activity. Although target heart rate remains one of the most effective instruments for monitoring exercise intensity, the rate of perceived exertion should be incorporated especially in the titration of exercise prescriptions for those on beta-blockade therapy. Finally the benefits of an exercise programme, derived from a foundation of proper assessment, are numerous and include improvements in cardiovascular fitness, body composition, blood lipid profile and retention of essential muscle mass during the course of the life-cycle. A considerable public health benefit will result if sedentary individuals become regularly more physically active. PMID- 8724202 TI - Benefits of aerobic exercise after stroke. AB - The debilitating loss of function after a stroke has both primary and secondary effects on sensorimotor function. Primary effects include paresis, paralysis, spasticity, and sensory-perceptual dysfunction due to upper motor neuron damage. Secondary effects, contractures and disuse muscle atrophy, are also debilitating. This paper presents theoretical and empirical benefits of aerobic exercise after stroke, issues relevant to measuring peak capacity, exercise training protocols, and the clinical use of aerobic exercise in this patient population. A stroke, and resulting hemiparesis, produces physiological changes in muscle fibres and muscle metabolism during exercise. These changes, along with comorbid cardiovascular disease, must be considered when exercising stroke patients. While few studies have measured peak exercise capacity in hemiparetic populations, it has been consistently observed in these studies that stroke patients have a lower functional capacity than healthy populations. Hemiparetic patients have low peak exercise responses probably due to a reduced number of motor units available for recruitment during dynamic exercise, the reduced oxidative capacity of paretic muscle, and decreased overall endurance. Consequently, traditional methods to predict aerobic capacity are not appropriate for use with stroke patients. Endurance exercise training is increasingly recognised as an important component in rehabilitation. An average improvement in maximal oxygen consumption (VO2max) of 13.3% in stroke patients who participated in a 10-week aerobic exercise training programme has been reported compared with controls. This study underscored the potential benefits of aerobic exercise training in stroke patients. In this paper, advantages and disadvantages of exercise modalities are discussed in relation to stroke patients. Recommendations are presented to maximise physical performance and minimise potential cardiac risks during exercise. PMID- 8724205 TI - Demonstration and distribution of HCV RNA sequences by in situ hybridization and HCV-related proteins by immunohistochemistry in the liver tissue of patients with chronic HCV infection. AB - Nonisotopic in situ cytohybridization of HCV RNA was attempted in liver specimens from 12 chronically hepatitis C virus (HCV) infected patients. Oligonucleotides deduced from 5'-noncoding and core regions of the HCV genome were labeled with digoxigenin and used on paraformaldehyde-fixed frozen liver sections. The hybrids were visualized immunohistochemically with alkaline phosphatase-conjugated anti digoxigenin and alkaline phosphatase substrate. These findings were correlated with the results of tissue immunohistochemistry for HCV antigens identified with specific mouse monoclonal antibodies developed against c22-3 antigen (Ag), a core encoded protein, and c100-3 Ag, a NS4-encoded protein, and histologic assessment of each liver. HCV RNA detected in the above assay was predominantly cytoplasmic; it was detected in all 12 patients and in none of the controls. Tissue HCV RNA was associated with the presence of cytoplasmic (c100-3 Ag) and membrane (c22-3 Ag) expression of viral proteins in all 9 patients with histological evidence of chronic progressive liver disease as judged by the presence of piecemeal necrosis, and lobular and portal tract inflammation. Despite the presence of abundant HCV RNA, none of 3 patients without histological evidence of chronic liver disease showed intrahepatocyte expression of viral proteins. These findings support the view that tissue HCV antigens are markers of progressive damage and demonstrate that active liver disease does not occur without such markers. It is proposed that synthesis of viral proteins and membrane accumulation of c22-3 Ag may be involved in the pathogenesis of hepatocyte injury in chronic hepatitis C infection. PMID- 8724204 TI - The role of quadriceps exercise in the treatment of patellofemoral pain syndrome. AB - In recent years there has been increased interest in the role of exercise in the alleviation of patellofemoral pain syndrome. Contradictions which result in a lack of a consensus amongst clinicians and researchers as to the most beneficial type of exercise are reviewed. The popularly held belief that there are imbalances between some components of the quadriceps femoris is also reviewed and conflicting evidence that quadriceps contractions can be enhanced by altering hip and knee positions is found. Some traditional exercises, previously considered essential to any regime for this common condition, were not found to be as beneficial as previously thought. Consequently, some of these exercises should be less prominent in a rehabilitation programme for patellofemoral pain syndrome. PMID- 8724206 TI - Expression of basic fibroblast growth factor and angiogenin in arthritis. AB - Angiogenesis is an integral component of the vasculoproliferative phase of rheumatoid arthritis (RA). We examined the distribution of two angiogenic factors, basic fibroblast growth factor (bFGF) and angiogenin (ANG) in arthritic diseases. We used an enzyme-linked immunosorbent assay and immunohistochemical analysis to determine the levels of bFGF and ANG in synovial fluid (SF) and their distribution in synovial tissue (ST), respectively. Most SF contained little or no detectable bFGF ( < 5 pg/ml). ANG in RA SF was 248.7 +/- 17.4 ng/ml, which did not differ significantly from levels found in osteoarthritis (OA; 305.9 +/- 23.1 ng/ml). Synovial lining cells, macrophages, endothelial cells, and vascular smooth muscle cells were immunopositive for bFGF and ANG; however, their expression was not up-regulated in RA ST compared to ST from OA and normal subjects. Though bFGF and ANG are present in the joints of patients with arthritic diseases, they are not up-regulated in RA. These results suggest that not all angiogenic mediators are up-regulated in RA compared to normal subjects and subjects with other arthritic diseases. It may be that some of these mediators, like ANG, play a role in the physiology of normal synovium. PMID- 8724207 TI - Expression of platelet-derived growth factor ligands and receptors by rat aortic endothelium in vivo. AB - In situ hybridization and immunostaining were used to study the time course of expression for platelet-derived growth factor (PDGF) ligands and receptors in endothelium of the rat aorta after injury. The PDGF-A and -B chains were expressed in endothelial cells at the wound edge within 4 h after injury, but no expression was detectable in uninjured endothelium. PDGF alpha-receptor was expressed in a pattern similar to the PDGF-A chain, while expression of PDGF beta receptor was not detected at any time. Expression of the PDGF-B chain remained elevated in endothelial cells at the leading edge even at later measurements when these cells had stopped replicating. Smooth muscle cells (SMCs), which are absent from the intima of the normal aorta and are known to express PDGF beta-receptors, were predominantly found to migrate into the intima near the endothelial leading edge where PDGF-B was expressed. These data suggest a paracrine role for endothelial PDGF in SMC migration. PMID- 8724208 TI - Quantitative reverse transcription polymerase chain reaction shows that ethanol enhances interleukin-6 mRNA expression in cultured mesangial cells. AB - IgA nephropathy, a form of mesangial glomerulonephritis, is associated with chronic ethanol ingestion in humans and in a rat model. We investigated the hypothesis that ethanol has a direct effect on a mesangial cell cytokine, interleukin-6 (IL-6). We measured IL-6 mRNA in cultured mesangial cells using a novel method, quantitative reverse transcription-polymerase chain reaction (Q-RT PCR). Primers were used to amplify a 346-bp segment. To quantify the results, we generated an internal standard using site-directed mutagenesis which resulted in a restriction site for EcoRI, absent in target IL-6 cDNA. In the presence of the same primers, the mutated internal standard (exogenous template) amplifies with equal efficiency as the target IL-6 cDNA. Q-RT-PCR, using 250 ng of total RNA, showed that after ethanol incubation, IL-6 mRNA was 65 (+/-30) attomol, a 1.5 fold increase from 44 (+/-28) attomol in control cells. This study shows that in vitro, ethanol enhances IL-6 mRNA expression in rat mesangial cells. PMID- 8724203 TI - Resistive exercise training in cardiac rehabilitation. An update. AB - Resistive exercise training has become very popular for patients of cardiopulmonary rehabilitation programmes (CRPs). For decades, CRPs focused almost exclusively on improving cardiorespiratory endurance and most programmes ignored muscular fitness development. Moreover, resistance training was thought to be potentially hazardous for the cardiac patient due to the risk of cardiovascular complications from adverse haemodynamic responses. We now know that resistive exercise testing and training is very safe for properly screened patients, even at relatively high workloads. Improvement in muscular strength facilitates return to daily vocational and avocational activities and is important for the CRP participant to regain lost strength and resume work soon after a cardiac event. Circuit weight training (CWT) is helpful in this respect and has been shown to increase muscular strength, cardiovascular endurance, body composition, bone density and mineral content, self-confidence, and self-efficacy in various populations. This article presents an update on current research in cardiac patients and also presents guidelines for implementing a properly supervised cardiac resistive exercise programme. PMID- 8724209 TI - Oxytetracycline-mediated alteration of murine immunocompetence. AB - The immunomodulatory effect of oxytetracycline (OTC) on murine splenic lymphocytes (MSL), peritoneal exudate macrophage (PEM) functions and antibody production was examined. In vivo exposure to OTC slightly delayed initiation of antibody formation during the primary response. However, OTC exposure had no effect on either the peak time of antibody response or peak antibody titer. OTC also had no significant effect on the secondary antibody response. Mitogen induced proliferation of MSL cocultured with OTC and pokeweed mitogen, phytohemagglutinin or concanavalin was equivocal. However, allogeneic stimulation of MSL was inhibited at 100 micrograms/ml OTC. There was also a decrease in the number of cells recovered. OTC had no effect on lymphocyte cytotoxicity in cells cultured in vitro. OTC inhibited the cytotoxic response of Corynebacterium parvum elicited PEM at 10 micrograms/ml (effector:target of 10:1). Low levels of OTC (1 10 micrograms/ml) augmented the cytotoxic response (effector:target of 5:1). The effect of OTC on induction of PEM cytotoxicity was assessed by coculturing thioglycollate-elicited (TG) PEM in vitro with IFN-gamma and endotoxin along with 0-100 micrograms/ml OTC. Induction of cytotoxicity was inhibited at 0.5 microgram/ml. The effect of OTC on TG-PEM antimicrobial activity was assessed by measuring reduction of nitroblue tetrazolium (NBT) and cytochrome C. OTC inhibited the reduction of NBT at 500 micrograms/ml following PMA stimulation of TG-PEM. OTC had no effect on either NBT or cytochrome C reduction following stimulation with opsonized zymosan. These results demonstrate that OTC-mediated immunosuppression is a multifaceted event, with differing sensitivities both between immune cells and between different pathways within the same cell. PMID- 8724210 TI - The action of phenazine methosulphate in causing cellular damage in the isolated mouse soleus muscle preparation. AB - Exposure of the isolated mouse soleus preparation to 1.0 mM phenazine methosulphate (PMS) caused (i) a slow and modest release of creatine kinase (CK) that was exacerbated by removal of extracellular Ca2+, (ii) a specific type of ultrastructural damage, namely a characteristic spacing of the myofibrils, (iii) swelling of the mitochondria, indicating a modest rise in [Ca2+]i, and (iv) swelling of the sarcoplasmic reticulum (SR). It is suggested that PMS (i) activates a sarcolemma oxidoreductase which synergistically interacts with raised [Ca2+]i to cause modest CK efflux and (ii) activates an oxidoreductase on the SR, thereby generating electrons which directly modify the integrity and organisation of the contractile apparatus. PMID- 8724211 TI - Effect of recombinant growth hormone and chromium picolinate on cytokine production and growth performance in swine. AB - The effect of dietary chromium picolinate (CrP) and recombinant porcine growth hormone, somatotropin (rPST) administration on growth performance and cytokine production in Landrace-Poland China gilts was determined using a 2 by 2 treatment array. Treatments were: (1) control (basal diet), (2) CrP-supplemented diet (basal diet + 300 micrograms Cr3+/kg diet as CrP), (3) rPST (100 pg/kg body weight/day), and (4) rPST+CrP. CrP-supplemented diets were fed beginning at 20 kg body weight through 90 kg. Administration of rPST was begun at 60 kg weight and continued through 90 kg. All rPST treated pigs demonstrated improvements in growth performance versus controls. Pigs given CrP-supplemented diets showed no differences in growth performance. At 90 kg, pigs were challenged with endotoxin (lipopolysaccharide, 0.2 microgram/kg i.v.). Blood samples were collected at 0, 1, and 3 h postchallenge. Plasma IL-6 levels increased from 23 U/ml at time 0 to 1,927 U/ml at 3 h for control swine. Swine from the CrP treatment group had IL-6 levels of 8,130 U/ml at 3 h post-LPS. There were no differences in plasma IL-6 from pigs in the rPST and rPST+CrP treatment groups compared to the controls. Endotoxin challenge had no effect on either blood glucose levels or induction of TNF-alpha in any treatment group. PBMC from CrP-treated animals produced more IL 2 than peripheral blood mononuclear cells from all other groups. PMID- 8724212 TI - Lack of natural killer cell augmentation in vitro by human interferon gamma in a subset of patients with systemic sclerosis. AB - Systemic sclerosis (SSc) is a generalized connective tissue disorder characterized by fibrosis of skin and various viscera. Natural killer (NK) cells are a subset of lymphocytes that can lyse targets without prior sensitization. Few studies have tried to assess NK cell function in patients with SSc. To evaluate NK cell cytotoxicity in patients with SSc and to see the extent of its augmentation in vitro by human interferon (hIFN) gamma in the clinical subset of limited and diffuse cutaneous diseases, we evaluated 27 patients with SSc and 22 age- and sex-matched controls by 51Cr release assay. Fifteen patients had limited cutaneous disease (mean disease duration 6.2 +/- 2.7 years) and 12 diffuse cutaneous disease (mean disease duration 5.7 +/- 2.4 years). Patients with limited SSc had significantly higher baseline NK cell function than controls (p < 0.05) and the augmentation following in vitro stimulation with hIFN gamma was negligible. Patients with diffuse SSc had lower baseline NK cell cytotoxicity than controls but this was not statistically significant. Augmentation with hIFN gamma in this group was comparable to controls. This study suggests that NK cells may have a role in the pathophysiology of this disease. PMID- 8724213 TI - Understanding research synthesis (meta-analysis). AB - Synthesis of research findings has long been a part of reviewing and summarizing a field of study. Public health decisions are made on the available evidence. We summarize the approaches to research synthesis that draw on the best available evidence and the use of quantitative summaries through meta-analysis. We focus on observational studies. Heterogeneity offers the potential to observe a relation across study populations and circumstances. We emphasize the benefits of heterogeneity in overviews and the need to explore and describe the sources of heterogeneity. Random effects approaches to combining data are recommended, and the use of regression approaches is emphasized. Excluding studies with extreme results may bias a research synthesis and underestimate the true variance of the results, thus contributing to misleading inference. Thorough searching is the best guard against publication bias. We conclude with guidelines for combining epidemiological studies. PMID- 8724214 TI - Disability as a public health outcome in the aging population. AB - Improvements in life expectancy in the twentieth century have resulted from major declines in mortality at younger ages, but it is less well recognized that mortality declines at older ages have also played a substantial role in prolonging expectation of life. A person reaching age 65 in 1900 could expect to live an additional 11.9 years. Life expectancy at age 65 rose to 14.4 years by 1960 and then increased by about three years in the next three decades, reaching 17.5 years in 1992 (56, 70). As a greater proportion of the population survives to very old ages, the public health impact of the burden of disease and disability and related utilization of medical care and need for supportive and long-term care has become an important concern. In particular, the ability of the older person to function independently in the community is a critically important public health issue. A growing body of research in the last decade has addressed the measurement of disability, factors related to its onset, consequences of disability, and the potential for preventive interventions. This article summarizes the state of the art in these areas and discusses their public health relevance. PMID- 8724215 TI - Epidemiology and prevention of breast cancer. AB - Breast cancer is the most common cancer in women, accounting for 32% of all newly diagnosed cancers. Demographic characteristics associated with an increased risk include increasing age, birth in North America or northern Europe, high socioeconomic status, never having been married, and, for breast cancer diagnosed after 45 years of age, the white race. Early age at menarche, late age at menopause, late age at first full-term pregnancy, and low parity increase risk, while removal of the ovaries at an early age is protective. Obesity increases risk in postmenopausal women. Having a first degree relative with breast cancer confers an increased risk, especially if both a mother and a sister have had breast cancer at an early age. Mutations in the BRCA1 and BRCA2 genes are associated with an inherited susceptibility to breast cancer at an early age. Other markers of increased risk include atypical and hyperplastic epithelial cells in nipple aspirate fluid, nodular densities on mammogram, and biopsy confirmed benign proliferative breast disease. Little can be done at present to reduce breast cancer risk through primary prevention, but secondary prevention by mammographic screening in women of age 50 and older reduces mortality from breast cancer. PMID- 8724216 TI - Cervical intraepithelial neoplasia, cervical cancer, and HPV. AB - Major progress has been made to control cervical cancer in the United States and Europe using screening programs, although it remains a major cause of morbidity and mortality in the developing world. The association between cervical cancer and a sexually transmissible etiologic agent was hypothesized long before identification of human papillomaviruses (HPV) as agents that infected the genital tract. HPVs are among the most common sexually transmitted agents and have been shown to induce several squamous anogenital cancers, including squamous cell cancer of the cervix. After an etiologic role for HPV was identified in cervical cancer and CIN, efforts to understand the molecular biology of HPV were greatly expanded, enhanced by the advent of deoxyribonucleic acid (DNA) probes to identify HPV infection. Recent research has focused on specific types of HPV in relation to other recognized risk factors in the pathogenesis of CIN and invasive cervical cancer. PMID- 8724217 TI - Ovarian cancer. AB - Ovarian cancer incidence rates are highest and stable in white populations; in Asia previously low incidence rates may be increasing. Most cases present with disseminated disease, and mortality rates remain high despite the use of aggressive polychemotherapy. Mortality among younger women has decreased, which has been attributed to widespread use of oral contraceptives. Studies consistently show a protective effect of oral contraceptives that increases with duration of use; no dose effect has been identified to date. Risk decreases substantially with increasing numbers of pregnancies; periods of lactation are relatively less protective. Periods of oral contraceptive use are less protective than equal periods of pregnancy. These factors may protect against ovarian cancer due to inhibition of ovulation or due to suppression of another aspect of ovarian function. Hysterectomy and tubal ligation are both protective, perhaps by preventing the ascent of environmental carcinogens that are as yet unidentified. A positive family history substantially increases risk; mutations in the BRCA1 gene may be responsible for about 5% of cases. No other exposures have been consistently associated with disease risk. Whether risk is modified by ovarian damage mediated by dietary galactose is being evaluated; studies to date have conflicting results. The effect of infertility and its treatments on ovarian cancer risk is controversial; two studies suggest that infertility treatments increase risk. PMID- 8724218 TI - Female lung cancer. AB - Female lung cancer death rates increased by more than 550 percent between 1950 and 1991. In 1986 lung cancer surpassed breast cancer to become the leading cause of cancer death in women in the United States. The lung cancer epidemic is primarily attributable to cigarette smoking, which is responsible for at least 80% of the disease in women. There are gender differences in the distribution of lung cancer by histologic type, even controlling for smoking, and some data suggest greater female than male susceptibility to lung cancer at a given level of smoking. Exposure to environmental tobacco smoke increases risk of lung cancer in nonsmoking women. Family history of lung cancer, personal history of lung disease (e.g. asthma, chronic bronchitis, pneumonia, or tuberculosis), and a history of radiotherapy also appear to be associated with increased risk. Data specific to women on the role of household radon exposures or of specific occupational or environmental exposures are relatively sparse and inconsistent. Finally, many studies have reported a decreased risk of lung cancer in individuals who consume high levels of fruits and vegetables; however, clinical trials fail to support a beneficial effect of beta-carotene supplementation. Since cigarette smoking accounts for the vast majority of lung cancer cases in women, efforts to prevent adolescent girls from starting to smoke and to encourage cessation among established smokers have the greatest potential for reducing the female lung cancer burden. PMID- 8724219 TI - Public health aspects of ophthalmic disease: introduction. PMID- 8724220 TI - The epidemiology and control of open angle glaucoma: a population-based perspective. AB - Chronic open angle glaucoma is an etiologically heterogeneous group of diseases characterized by damage to the optic nerve, resulting in peripheral visual loss that can progress to involve the fovea and central vision. Open angle glaucoma can be divided into primary conditions and conditions which are secondary to another ocular or systemic disease. Causes of secondary glaucoma include uveitis, cataract, trauma, and disorders affecting the development structure of the angle. This review focuses on primary open angle glaucoma (POAG), since it accounts for the vast majority of the disease burden in the US population and its etiology remains unknown. PMID- 8724221 TI - Diabetic retinopathy. AB - Diabetic retinopathy is a leading cause of loss of vision in the United States. Results of recent population-based studies and randomized controlled clinical trials suggest that glycemic control can lower the incidence and prevent the progression of retinopathy and loss of vision associated with diabetes. In addition, data from clinical trials showed that timely photocoagulation treatment of severe proliferative retinopathy or clinically significant macular edema prevents loss of vision. This report reviews the epidemiology of diabetic retinopathy and highlights areas in need of further epidemiologic research. PMID- 8724222 TI - Blindness due to cataract: epidemiology and prevention. AB - Cataract is the leading cause of blindness, accounting for 50% of blindness worldwide. Although significant progress has been made toward identifying risk factors for cataract, there is no proven primary prevention or medical treatment. Surgical removal of cataract remains the only therapy. In this paper, we reviewed (a) the prevalence and incidence data of cataract, (b) the findings on risk factors for cataract, and (c) health service utilization and outcome of cataract surgery in both developed and developing countries. PMID- 8724223 TI - Public health and regulatory considerations of the Safe Drinking Water Act. AB - This paper provides an overview of the public health and economic issues associated with drinking water quality regulations in the United States. A historic perspective is provided by the use of filtration and chlorine disinfection, and of public health laws from the early 20th century up to passage of the Safe Drinking Water Act (SDWA), in 1974. The contaminants regulated under the Act, and the 1986 Amendments to the SDWA, are evaluated according to health endpoint, related issues in risk assessment, and the cost of complying with associated regulations. Risk-cost and benefit-cost analyses are offered for carcinogens, systemics, and pathogens. The paper describes the evolution of public health issues from the initial focus on waterborne infectious diseases to concerns over chemical contaminants, and the recent reemergence of microbials as the high-priority public health concern. PMID- 8724224 TI - Benefit-cost analysis in public health. AB - This chapter gives an overview of benefit-cost analysis (BCA) and similar evaluation tools, such as cost-effectiveness analysis and technology-based standards, in the context of public health. We describe these evaluation tools, how they are used, their shortcomings, and how they should be interpreted. As with other powerful tools, they are subject to misuse and misinterpretation, even by professionals doing the analysis. PMID- 8724225 TI - Health hazards to humans associated with domestic pets. AB - Allergy to pets, particularly when manifested as asthma, is an important health hazard. The health impact of severe clinical disease associated with allergy to pets can be minimized by avoidance of pets. Many individuals with milder disease keep their pets and obtain medications for relief of their allergy symptoms. Bird fancier's disease can result in profound deterioration of pulmonary function, and requires avoidance of bird exposure. Animal bites are the commonest health hazard of domestic pets and result in tissue damage and infection risk. Bites are treated by cleaning the wound, and appropriate use of antibiotics and tetanus prophylaxis. Diverse infectious diseases are transmitted to humans by their pets; these diseases are particularly dangerous to immunocompromised individuals. The infectious diseases can be partly prevented by avoiding ill animals and by washing of hands following exposure to pets or pet-derived secretions. They can be treated by appropriate antibiotics and/or specific immune globulins and/or vaccines. PMID- 8724226 TI - Health enhancement and companion animal ownership. AB - The relationship between people and companion animals, on the one hand, explains the bites and zoonotic diseases that occur among those with companion animals and, on the other hand, appears to enhance the psychological and physiological well-being of many people. Presently, no less than 56% of households in the United States have animals, typical of developed countries around the world. It is well documented that people denied human contact do not thrive well. All indications are that companion animals play the role of a family member, often a member with the most desired attributes. Animals play special roles for children, aiding the teaching of nurturing behavior and appreciation of nonverbal communication. Ordinary interactions with animals can reduce blood pressure and alter survival after a heart attack. For some, pets increase the opportunities to meet people, while for others pets permit them to be alone without being lonely. PMID- 8724227 TI - Crack cocaine abuse: an epidemic with many public health consequences. AB - In the mid-1980s a new, smokable form of cocaine, called crack, was introduced in the United States. Soon thereafter, it became apparent that crack cocaine abuse was a serious and important public health concern. Over the past several years, crack cocaine use has increasingly been associated with a myriad of immediate and long-term adverse effects. During this same period, crack cocaine use has progressively moved away from experimentation and recreational use to chronic and compulsive drug use. PMID- 8724228 TI - Pervasive media violence. AB - In this review, we focus our discussion on studies examining effects on children and young adults. We believe that the current epidemic of youth violence in the United States justifies a focus on this vulnerable segment of society. We consider media effects on individual children's behaviors, such as imitating aggressive acts. In addition, we examine how the media influence young people's perceptions of norms regarding interpersonal relationships. Next, we assess mass media effects on societal beliefs, or what children and adolescents think the "real world" is like. We suggest these media influences are cumulative and mutually reinforcing, and discuss the implications of repeated exposure to prominent and prevalent violent media messages. Finally, we catalog multiple intervention possibilities ranging from education to regulation. From a public health perspective, therefore, we evaluate the effects that pervasive media messages depicting violence have on young people and present multiple strategies to promote more healthful outcomes. PMID- 8724229 TI - Healthy Cities: toward worldwide health promotion. AB - Healthy Cities is a community problem-solving process for health promotion that began in Canada and Europe in the mid-1980s. Although Healthy Cities have expanded worldwide, there is limited scientific literature that documents the effects of these efforts. This review describes the dynamic status of Healthy Cities globally and summarizes what is known about these efforts. It is difficult to draw generalizations or clear differences between Healthy Cities and other efforts with similar objectives in the United States. There appears to be less variation among Healthy Cities in Europe, most likely because of the extensive technical support provided by the WHO European Regional Office to the Healthy Cities Project. Case descriptions present the diversity and types of activities of Healthy Cities. It is concluded that community participation and broad involvement from different sectors of the community are promising approaches to addressing urban problems. PMID- 8724230 TI - Mental health problems of homeless women and differences across subgroups. AB - Homeless women are a large and diverse group, constituting one fifth of the US homeless adult population. Although most homeless women do not have major mental illness, homeless women exhibit disproportionately high rates of major mental disorders and other mental problems. Rates of mental disorders are highest among whites and women without children, and important variations by subgroups of homeless women reinforce the need for disaggregated analysis. Many homeless women with serious mental illness are not receiving needed care, apparently due in part to the lack of perception of a mental health problem and the lack of services designed to meet the special needs of homeless women. PMID- 8724231 TI - Community-based approaches for the prevention of alcohol, tobacco, and other drug use. AB - This paper summarizes what is known about community-based approaches for the prevention of ATOD problems and how the current practices in the field reflect these approaches. The first section of the chapter provides a brief summary of events early in this century when community-based approaches were central to addressing alcohol and other public health problems. The second section contains an overview of current research and empirical findings that yield consensus as to what conceptually and in practice constitutes a comprehensive, community-based prevention program for the prevention of ATOD problems. The third section reviews the literature of existing programs to assess the extent to which they include the salient elements and employ interventions determined to be fundamental to comprehensive community-based prevention programs. The final section discusses some of the challenges that confront researchers and practitioners when developing prevention initiatives and programs in high-risk environments. PMID- 8724232 TI - Population and women's reproductive health: an international perspective. AB - This paper gives a brief overview of current world population or demographic issues, followed by a discussion of the ICPD proceedings and various notable aspects of the ICPD Programme of Action. It then focuses on six of the most pressing reproductive health concerns facing women today: gender inequalities, access to contraceptive services, sexually transmitted diseases (including HIV), maternal mortality, unsafe abortion, and adolescent pregnancy. Because the ICPD Programme of Action is intended to have far-reaching consequences for each of these issues, it is taken as a focal point of analysis. PMID- 8724233 TI - The role of medical examiners and coroners in public health surveillance and epidemiologic research. AB - The role of medical examiners and coroners (ME/Cs) in public health surveillance and epidemiologic research is reviewed. Definitions are offered, and discussion centers on the advantages of, and obstacles to the use of ME/C data; existing surveillance systems relevant to ME/Cs; studies assessing the usefulness of ME/C data; newly emerging tools for ME/C surveillance and epidemiologic research; and recommendations for the future. ME/C data have been used quite successfully in some settings and are potentially very useful for surveillance and epidemiologic research on a large scale, but the data have limitations that need to be addressed in the future. PMID- 8724234 TI - Racial differences in health: not just black and white, but shades of gray. AB - Explanations for racial/ethnic disparities in health are varied and complex. This paper reviews the literature to assess the extent to which current disparities are a consequence of racial differences in the social class composition of the US population. We focus this review on African Americans and examine studies that provide information on the effect of race on four outcome measures: infant mortality, hypertension, substance use, and mortality from all-causes. Twenty three studies were identified that met criteria for inclusion in this review. As expected, most studies provide evidence that socioeconomic conditions are a major factor explaining racial differences in health. Findings, however, vary for the different health indices. Research in the area of substance abuse provides the most consistent evidence that socioeconomic conditions account for observed racial differences. In contrast, studies on infant mortality and hypertension provide a compelling case that the effects of socioeconomic status are important, but not sufficient to explain racial differences. Evidence on mortality from all causes is equally divided between studies showing no significant race effect and those in which racial differences persist after adjusting for social class. The paper offers possible explanations for the seemingly divergent results and identifies conceptual and methodologic issues for future research seeking to disentangle the complex relations between race, social class, and health. PMID- 8724235 TI - Health and unemployment. AB - This paper reviews the relationship between health and inadequate employment, especially unemployment. Poor physical or mental health can lead, via poor work performance, to job loss; however, studies that control for such selection effects are still scarce except for a few health outcomes. For example, aggregate level studies typically find a positive association between unemployment and suicide rates over time. At the individual level of analysis, panel surveys of laid-off workers tend to find increased psychiatric problems such as depression and substance abuse. Few studies have evaluated interventions to prevent or reduce the adverse health effects of job loss. There have been even fewer studies of the health effects of other types of inadequate employment such as the increasingly prevalent forms of underemployment. PMID- 8724236 TI - Economic evaluation of HIV prevention programs. AB - Program managers and policy makers need to balance the costs and benefits of various interventions when planning and evaluating HIV prevention programs. Resources to fund these programs are limited and must be used judiciously to maximize the number of HIV infections averted. Economic evaluation studies of HIV prevention interventions, which we review and critique here, can provide some of the needed information. Special emphasis is given to studies dealing with interventions to reduce or avoid HIV-related risk behaviors. Ninety-three cost benefit, cost-effectiveness and cost-utility analyses were identified overall. However, only 28 dealt with domestic, behavior change interventions; the remainder focused on screening and testing without prevention counseling, and on care and treatment services. There are compelling demonstrations that behavioral interventions can be cost-effective and even cost-saving. The threshold conditions under which these programs can be considered cost-effective or cost saving are well defined. However, several important intervention types and multiple key populations have been unstudied. Research in these areas is urgently needed. PMID- 8724237 TI - Prevention and the reforming U.S. health care system: changing roles and responsibilities for public health. AB - This review presents historical and cost-effectiveness perspectives of prevention in health care; discusses the nature, extent, and determinants of health system change, particularly the transition to managed care with large integrated health care corporations; and identifies implications for public health agencies and opportunities for prevention within the reforming health system. PMID- 8724238 TI - Developing evidence-based clinical practice guidelines: lessons learned by the US Preventive Services Task Force. AB - The US Preventive Services Task Force is an expert panel established by the federal government in 1984 to develop evidence-based practice guidelines on screening tests and other preventive services. Its recommendations are published elsewhere. This article explores the lessons learned in the process of developing and disseminating the recommendations. Topics include project organization (analytic philosophy, project sponsorship, panel composition, topic selection); the review of evidence (selecting outcome measures for judging effectiveness, constructing "causal pathways," searching the literature, rating the evidence, synthesizing the results); crafting recommendations (extrapolation, assessing magnitude, balancing risks and benefits, addressing costs, dealing with insufficient data, separating science from policy); peer review; collaboration with other groups; evaluating impact on clinicians' knowledge, attitudes, and behavior; updating recommendations; and defining a research agenda. The lessons learned suggest potential refinements in the future work of the task force and other groups engaged in guideline development. PMID- 8724239 TI - Cell surface hydrophobicity and medically important fungi. PMID- 8724240 TI - Nomenclature of fungal diseases. PMID- 8724241 TI - Treatment of eumycetoma and actinomycetoma. AB - Mycetoma is a chronic disease caused by aerobic actinomycetes and eumycetes which mainly affects the lower extremities. It predominates among farm workers in tropical, subtropical and adjacent zones. Clinically it is characterized by a firm swelling with abscesses and fistulae discharging pus that contains granules or grains of the causal agent. Their color, size, consistency and histopathology contribute to their identification. Cultures and metabolic studies determine the disease's etiology. Eumycete and actinomycete antigens can be used serologically to diagnose and predict prognosis of the disease. Many different antimicrobials and antifungal drugs have been used with varying degrees of success. Trimethoprim sulfamethoxazole alone or together with diamino-diphenyl-sulfone is the treatment of choice for actinomycetoma. Amikacin is used for severe cases, unresponsive to previous treatment, and for those in danger of dissemination to adjacent organs. Surgery is seldom used for actinomycetoma. In eumycetoma a combination of medical treatment and surgery is advised. Small eumycetomas are easily surgically removed. Ketoconazole at a dosage of 400 mg/day is the medical treatment of choice for eumycetoma caused by M. mycetomatis. The therapeutic response to itraconazole varies. Fluconazole has been unsuccessful in the treatment of eumycetoma but amphotericin B has shown good to poor therapeutic response. PMID- 8724242 TI - A contemporary view of sporotrichosis. PMID- 8724243 TI - Seborrhoeic dermatitis and Pityrosporum yeasts. AB - The connection between P. ovale and seborrhoeic dermatitis has been clearly demonstrated in a number of treatment studies but we still do not know how P. ovale induces skin lesions. An enhanced growth of P. ovale cannot be the cause, because a number of studies with quantitative determinations of P. ovale have not been able to show any difference in the number of yeast cells between patients and healthy controls. The number of P. ovale is probably only important for the individuals who are susceptible to seborrhoeic dermatitis. An abnormal immune response to P. ovale could be another explanation. Sohnle et al. have shown that P. ovale can activate complement by both the classical and the alternative pathway. A defective cell-mediated immunity to P. ovale in patients with seborrhoeic dermatitis has been demonstrated by Wikler et al. In patients with AIDS, who are known to have a diminished T-cell function, a high incidence of seborrhoeic dermatitis has been found. Activation of the alternative complement pathway by P. ovale, which does not require T-cell function, could be an explanation for the inflammatory response. I also believe that the skin lipids are important in the pathogenesis. An improvement of seborrhoeic dermatitis has been demonstrated after treatment with drugs that reduce the sebum excretion. Pityrosporum has lipase activity and may generate free fatty acids, which could also contribute to the inflammatory response. There are a number of factors which are probably important in the pathogenesis of seborrhoeic dermatitis, that is, the number of P. ovale, P. ovale lipase activity, skin lipids, immune function, heredity, atmospheric humidity and emotional state. A reduction in the number of P. ovale in patients suffering from seborrhoeic dermatitis and being treated with antimycotic treatment is, at the present state of knowledge, the best way to treat the disease. PMID- 8724244 TI - Oral candidiasis in HIV-infected patients. PMID- 8724245 TI - Immunology of fungal infection. PMID- 8724246 TI - Cell wall polysaccharides of pathogenic yeasts. PMID- 8724247 TI - Chromomycosis. PMID- 8724248 TI - Tools and trends in the detection of Aspergillus fumigatus. PMID- 8724249 TI - Dematiaceous fungal infections in China. PMID- 8724250 TI - Excluding gastroesophageal reflux disease as the cause of chronic cough. AB - Gastroesophageal reflux disease (GERD) is recognized to be present in 10-20% of cases of chronic cough. Proving that it is the cause of the cough is more difficult. This problem is illustrated by way of a case report demonstrating that GERD can still be the cause, even when the patient is unresponsive to conventional use of proton pump inhibitors. In the commentary following the case history, we review the medical literature to confirm that GERD and cough may each precipitate the other. The role of esophageal pH monitoring in difficult cases of chronic cough is explored; we emphasize the use of pH monitoring while the patient is on therapy to prove or disprove the link. PMID- 8724251 TI - Genomania of p53 protein in gastric cancer. AB - The role of oncogenes and tumor suppressor genes in the pathogenesis of gastric cancer has recently received considerable attention. p53 is a tumor suppressor gene that is essential in the cell cycle; it prevents G1/S phase transition, after exposure to ionizing radiation or DNA-damaging chemotherapy. This allows the cell to repair its DNA or, if the damage is irreversible, to elicit apoptotic cell death. p53 mutations are seen in many human tumors including gastric carcinoma. Evidence suggests that mutant p53 is associated with shorter life expectancy in gastric, breast, lung, and colorectal cancer. A number of studies have shown cellular resistance to chemotherapy in the presence of mutant p53. Currently, increasing interest has been devoted to the potential role of mutant p53 as a screening tool. PMID- 8724252 TI - Role of dental plaque in the transmission of Helicobacter pylori infection. AB - With regard to the role of dental plaque in the transmission of Helicobacter pylori infection, data from the literature vary greatly, owing to differences in sample collection and H. pylori-detecting techniques. Using the polymerase chain reaction (PCR), we have determined the incidence of H. pylori colonization in the dental plaque of 31 consecutive patients who underwent gastroscopy. The patients were divided into two groups on the basis of H. pylori infection, determined by Giemsa stain and the rapid urease test: group A made up of 21 H. pylori-positive patients and group B with 10 H. pylori-negative patients. Our PCR assay of dental plaque samples proved negative in all group A subjects but was positive in only one patient in group B. In our study, we found that H. pylori had a low prevalence (3.2%) in the oral cavity, with no significant relationship between gastric mucosa and dental plaque colonization. More comprehensive studies are needed to determine whether dental plaque is an important reservoir in the epidemiology of H. pylori-induced gastric disease. PMID- 8724253 TI - Are duodenal ulcer seasonal fluctuations paralleled by seasonal changes in 24 hour gastric acidity and Helicobacter pylori infection? AB - The occurrence of duodenal ulcer is characterized by seasonal variation, for poorly understood reasons. No previous study has assessed whether 24-h intragastric acidity and Helicobacter pylori infection have similar seasonal fluctuations in patients with this disorder. For this reason, we evaluated retrospectively the circadian gastric pH in 319 new patients (226 men and 93 women, mean age 45.2 years) with endoscopically proven duodenal ulcer, who agreed to undergo this examination during the years 1987-1992 in our center. The month by-month occurrence of the disease over the global 6-year period was assessed, and the mean pH values were calculated for each patient during three time intervals of interest: 24 h, daytime (08:00-19:59 h), and nighttime (20:00-07:59 h). The mean pH values of these three time periods were then calculated month by month throughout the annual cycle. H. pylori infection was sought by histology in 171 patients examined in the period from 1990 to 1992. The percentage of H. pylori-positive duodenal ulcer patients was then calculated for each season. The calendar fluctuation of duodenal ulcer occurrence showed an evident increase (p < 0.001) in fall (October-December) and in winter (January-March) compared with spring (April-June) and summer (July-September). Both 24-h and nighttime gastric acidity showed no significant variation by month, whereas daytime gastric pH varied significantly (p < 0.05) with two evident decreases, meaning higher acidity, in April and August. H. pylori infection was detected in 152 of 171 patients (89%), and the percentage of H. pylori-positive duodenal ulcers did not differ from season to season. We conclude that there was no parallel circannual fluctuation of duodenal ulcer, gastric acidity, and H. pylori infection in the restricted sample of patients we studied. This reduces the apparent relevance of acid in inducing ulcer seasonal fluctuation. Also, the responsibility of H. pylori in this phenomenon can be excluded until a reliable diagnostic method capable of distinguishing recent from old infection is found. PMID- 8724254 TI - Helicobacter pylori serology in chronic gastritis with antral atrophy and negative histology for Helicobacter-like organisms. AB - It has been suggested that there may be a correlation between Helicobacter pylori (Hp) infection and precancerous lesions of the stomach. However, histological evaluation of bacterial colonization in chronic atrophic gastritis shows a relatively low prevalence of the microorganism, which does not support the hypothesis. The aim of our study was to investigate the Hp serology in 95 patients with chronic gastritis with antral atrophy, with (27 cases) and without (68 cases) intestinal metaplasia, and without Helicobacter-like organisms in antral and corpus biopsy specimens. For all subjects, serum anti-Hp immunoglobulin IgG was identified by a fluorescent immunoenzymatic method (Helori test; Eurospital), and mucosal atrophy and activity were graded histologically (Sydney System score). The serum Hp-antibody status documented the presence of current bacterial infections in 64 of 95 (67.4%) patients and previous infections in another 17 subjects. In only 14.7% of cases was there no evidence of current or previous infection. These subjects had less severe mucosal atrophy and lower inflammatory scores. In addition, there were no cases of intestinal metaplasia in such subjects. The high prevalence of Hp infection confirms the primary role of the microorganism in the pathogenesis of chronic gastritis with antral atrophy, although the bacterium is no longer present in the advanced stages of such disease. The histological evaluation of Hp colonization following the criteria of the Sydney System appears from our study to underestimate the true prevalence of the infection in the stomach when there is mucosal atrophy. PMID- 8724255 TI - Whole-bowel irrigation as an adjunct to the treatment of chronic, relapsing Clostridium difficile colitis. AB - We report the successful treatment of two patients with chronic, intractable Clostridium difficile infection using whole-bowel irrigation with a polyethylene glycol solution (Golytely) as adjunctive therapy. Before this treatment, both patients had recurrent symptoms of diarrhea, weight loss, abdominal pain, and documented C. difficile toxin-positive stools despite multiple pharmacologic treatments. Each child was prescribed myriad drug therapies, including vancomycin, metronidazole, bacitracin, and rifampin. Cholestyramine and lactobacillus were also tried alone and in combination with antibiotics. In each case, symptoms recurred shortly after cessation of therapy. Whole-bowel irrigation was subsequently administered until profuse, clear liquid stools were produced. This treatment was followed by a 3-week course of oral vancomycin and lactobacillus. In both cases, the patient became asymptomatic within 3 days of therapy; they have remained symptom-free for 36 and 48 months, respectively. We suggest that whole-bowel irrigation clears active C. difficile organisms, toxins, and spores from the intestine and is effective as an adjunct to routine therapy for chronic, relapsing C. difficile infections. PMID- 8724256 TI - Chronic idiopathic constipation: pathophysiology and treatment. AB - Chronic constipation is common in the general population, especially in women, in its idiopathic form. However, confusion still surrounds its definition, despite recent efforts to standardize it. Constipation can be divided in two large subgroups-normal transit and slow transit. The have different pathophysiological bases still not completely understood. Most patients respond to simple therapeutic measures aimed at correcting dietary fiber intake and lifestyle. Others, however, need more aggressive treatment, including laxatives, psychological therapy, and biofeedback. In a few patients with intractable constipation, surgery might be indicated to give relief. PMID- 8724257 TI - The postcholecystectomy syndrome. A role for duodenogastric reflux. AB - We have assessed the relationship between dyspepsia and gallstones and evaluated the effects of cholecystectomy on symptoms, endoscopic findings, and degree of duodenogastric reflux. Thirty patients with gallstones were enrolled in our study. Their symptoms, gastroscopic findings, and bile salt concentrations in fasting gastric juice were evaluated before and after surgery. Before cholecystectomy, biliary colic was present in 26 patients and dyspepsia in 20 patients; 16 patients also had biliary colic. After surgery, biliary colic disappeared in all patients. Dyspeptic symptoms improved in 12 patients (40%), 13 (43%) remained the same, and five patients (17%) developed dyspepsia or showed increase in their symptoms, the postcholecystectomy syndrome (PCS). Endoscopic gastritis developed in 50% after surgery compared with 30% before. Benign gastric ulcers developed in three patients, whereas none had been present before. Concentration of bile salts in fasting gastric juice increased from 0.56 +/- 0.4 mM to 1.47 +/- 0.75 mM after cholecystectomy (p < 0.0001). There was a positive correlation between the severity of symptoms in the postcholecystectomy syndrome and the change in the concentration of bile salts in fasting gastric juice (p = 0.0012). These observations suggest that duodenogastric reflux may play a significant role in the pathogenesis of symptoms in the postcholecystectomy syndrome. PMID- 8724258 TI - Impaired pancreatic function in patients with Crohn's disease with and without pancreatic autoantibodies. AB - Pancreatic autoantibodies (PABs) are found in 31% of patients with Crohn's disease (CD), but they do not correlate with the activity of intestinal disease or the incidence of acute pancreatitis. Exocrine pancreatic insufficiency has been observed in patients with CD. The aim of our study was to correlate the occurrence of PABs with exocrine pancreatic function to explore the clinical significance of these antibodies. Serum samples of 64 patients with CD were tested for PABs by indirect immunofluorescence. In addition, all patients were tested for exocrine pancreatic insufficiency by a fluorescein dilaurate test. PABs were detected in 26 of 64 patients (40%). The PAB-positive and -negative groups did not differ in clinical characteristics, such as age, sex, involvement of intestine, previous surgical interventions, drug therapy, and disease activity. Seven of the antibody-positive patients (27%) had impaired pancreatic function, in contrast to three of 38 PAB-negative patients (8%) (p < 0.05). In conclusion, exocrine pancreatic function is impaired significantly more often in PAB-positive than in PAB-negative patients. A prospective follow-up is required to determine whether PAB-positive patients are more likely to develop pancreatic insufficiency later in their course of disease. PMID- 8724259 TI - Large-volume paracentesis in the treatment of cirrhotic patients with refractory ascites. The role of postparacentesis plasma volume expansion. AB - Ascites is a common complication of cirrhosis and has a major clinical impact on the patient's general well-being. Approximately 10% of patients with cirrhosis can develop diuretic-resistant, tense ascites that requires other therapeutic interventions. In recent years, there has been a renewed interest in large-volume paracentesis (LVP) as a safe, simple, and inexpensive method to substitute for other more complicated and costly therapeutic interventions for refractory ascites. In this article, we review the latest literature supporting the use of LVP for the treatment of refractory, tense ascites. We also address the role of intravascular volume expansion after LVP, note that usually no postparacentesis volume expansion is necessary, and compare, when used, the different plasma volume expanders in terms of efficacy, safety, and cost. PMID- 8724260 TI - Hepatotoxicity due to antituberculosis therapy. Clinical profile and reintroduction of therapy. AB - The clinical profile of antituberculosis treatment (ATT)-induced hepatotoxicity is variable, and the reintroduction of ATT in patients who have developed such injury is controversial. We conducted a prospective study to determine the clinical profile in patients with ATT-induced hepatotoxicity and to test a predefined strategy of reintroduction of ATT. Seventy-two consecutive patients with clinical evidence of ATT-induced hepatotoxicity were included. Jaundice was the presenting symptom in 44 (61%) patients; prodromal symptoms were present in 28 (39%). Serious complications developed in 12 (16.6%) patients (fulminant hepatic failure in seven, subacute hepatic failure in four, hepatic encephalopathy in one). Nine patients (three males, six females) died from these complications. The mean duration of treatment before the onset of hepatitis was significantly longer in the group that died (53.22 +/- 36.22 days) than in the rest of the patients (31.07 +/- 30.30 days; p < 0.01). Malnutrition was present in 37 of the 72 patients. After resolution of drug induced hepatitis, reintroduction of isoniazid and rifampicin was possible in 41 of 44 patients. Thus, our results showed that ATT-induced hepatitis carried significant morbidity and mortality, that malnutrition was common in patients with ATT-related hepatitis, and that potentially hepatotoxic antituberculosis agents could be safely reintroduced after recovery from hepatitis. PMID- 8724261 TI - Jejunal phlebectasia presenting with massive gastrointestinal hemorrhage. AB - Our case report illustrates the clinicopathologic features of this rare vascular lesion and highlights that phlebectasia should be considered as a cause of gastrointestinal bleeding of undetermined etiology in adult patients. PMID- 8724262 TI - A parastomal hernia causing small-bowel obstruction. AB - A parastomal hernia is an incisional hernia that occurs at the site of an intestinal stoma on the abdominal wall. We report the case of a 52-year-old woman who had had ulcerative colitis and a total colectomy and ileostomy 9 years before being evaluated for a 2-year history of intermittent small-bowel obstruction thought initially to be due to Crohn's disease. On examination, a clear parastomal hernia was detected and confirmed by a small-bowel series. At surgery the hernia was corrected, and her intermittent episodes of small-bowel obstruction resolved. A parastomal hernia should be considered as a cause of intermittent small-bowel obstruction in anyone who has a stoma. Careful history and physical examination should suffice in establishing a diagnosis. PMID- 8724263 TI - Fulminant herpes colitis in a patient with Crohn's disease. AB - Herpes simplex virus (HSV) is a well-recognized cause of gastrointestinal infection, most commonly in patients with underlying immunodeficiency. The esophagus, perianum, and rectum are the most common sites of involvement; however, extensive colitis is rare. We describe a woman with Crohn's disease who developed pathologically proven HSV colitis. We review the literature and present the possible implications of the diagnosis. PMID- 8724264 TI - Colonic ulceration associated with the systemic use of nonsteroidal antiinflammatory medication. AB - We report the first case of colonic ulceration precipitated by the intramuscular administration of ketorolac. This report suggests a role for the systemic effects of nonsteroidal antiinflammatory agents in the development of gastrointestinal ulceration. PMID- 8724266 TI - Clinical enteroscopy. AB - We prospectively evaluated the diagnostic yield of push enteroscopy using either the Olympus SIF 10-LY fiberscope or the SIF-100 video enteroscope in 120 patients suspected of having small-bowel pathology. Insertion beyond the ligament of Treitz, checked by fluoroscopy, ranged from 30 to 150 cm (median 100). The source of bleeding was identified in 25 (41%) of 61 patients referred for gastrointestinal bleeding of obscure origin; vascular malformations were the most common finding, followed by neoplasias. Of 16 patients with inherited polyposis syndromes, multiple distal duodenal or jejunal hamartomas were diagnosed in five of six with Peutz-Jeghers syndrome and multiple duodenal or jejunal adenomas in six of 10 with familial adenomatous polyposis. Overall, isolated vascular malformations were successfully cauterized in three patients, and polypectomy was performed in 12 patients. Endoscopic and histological alterations, in both duodenum and jejunum, were found in 16 of 43 patients with diarrhea and/or malabsorption of unclear origin. The remaining 27 had normal duodenal and jejunal mucosal appearances; six had abnormal histological findings only in the jejunum and one both in the duodenum and in the jejunum. If we take jejunal histology as diagnostic, the sensitivity of duodenal histology was 74% and the specificity was 100%. We conclude that push enteroscopy provides useful diagnostic information and has therapeutic capabilities in patients with small-bowel pathology. PMID- 8724265 TI - Antral nodularity, gastric lymphoid hyperplasia, and Helicobacter pylori in adults. AB - Our objective is to evaluate endoscopically-diagnosed antral nodularity in adults and its relationship to lymphoid hyperplasia (LH) and Helicobacter pylori (HP). Antral biopsy specimens were studied for inflammation, presence of HP, and lymphoid follicles. Patients with marked functional dyspepsia or recurrent duodenal ulcers were given triple therapy (bismuth subcitrate, tetracycline, and metronidazole in the recommended doses for 2 weeks) to eradicate HP. Follow-up endoscopy and biopsies, at least 4 weeks after finishing treatment, were performed to assess eradication of HP and its effect on nodularity and LH. In all 25 patients (age range, 20-42 years) with antral nodularity, biopsy specimens were positive for HP. Twenty (80%) of subjects had lymphoid follicles; 13 of these 20 were given triple therapy. Eradication of HP was achieved in five cases (38%). Patients in whom HP was successfully eradicated showed improvement of their symptoms; antral nodularity subsided and there was marked regression of the lymphoid follicles. Antral nodularity with LH, reported to be unique to children, is not uncommon in adults and is induced by HP, eradication of which leads to regression of nodularity and LH in most cases. Low eradication rates achieved with metronidazole-based triply therapy is due, possibly, to primary resistance to metronidazole. Long-term follow-up of such patients is required to assess the evolution of these findings. PMID- 8724267 TI - Endoscopic treatment of ampullary adenomas in familial adenomatous polyposis. AB - Duodenal adenomas, usually considered premalignant, are found in < or = 100% of patients with familial adenomatous polyposis (FAP). Endoscopic screening is accepted, but the optimal treatment is unclear. Our objective was to assess endoscopic treatment of the upper gastrointestinal tract in patients with FAP. We reviewed the clinical records of 393 FAP patients in detail. Six patients had ampullary cancers. Sixty-nine had periampullary adenomas, none of whom developed malignancy during follow-up. Several endoscopic approaches were used, leading to various outcomes. (a) Follow-up with ampullary biopsy was the only method in 18 patients, with macroscopic improvement in one, unchanged condition in 11, and enlargement of adenomas in six. (b) Thermal ablation was used in 19 patients, with resolution in 10, improvement in seven unchanged condition in one, and one recurrence. (c) Yearly push enteroscopy, duodenoscopy, and ampullary biopsies were conducted in 11 of the 19 patients treated first with thermal ablation. Positive biopsies resulted in endoscopic retrograde cholangiopancreatography (ERCP), prophylactic sphincterotomy, and ablation with reexamination every 2-6 months. Follow-up of the patients treated with this last and favored strategy showed that five experienced resolution of symptoms, five had macroscopic improvement, and one had macroscopic as well as histologic progression. We conclude that patients with FAP should have periampullary surveillance, including duodenoscopy and biopsies from the time of diagnosis. Periampullary adenomas can be eradicated endoscopically. It is not clear whether ablation of adenomas or periodic biopsy is the ideal treatment. PMID- 8724268 TI - Anaplastic carcinoma arising in ectopic pancreas located in the distal esophagus. PMID- 8724269 TI - Colonic carcinoma after duodenocolic anastomosis. PMID- 8724270 TI - Mycobacterium avium complex peritonitis in a patient with alcoholic liver disease. PMID- 8724272 TI - Rigid bronchoscopy. AB - Rigid bronchoscopy is indicated for the presence of significant bleeding, a foreign body, or in pediatric patients. A thorough knowledge of endobronchial anatomy and expertise with rigid bronchoscopy is a necessity for the practicing general thoracic surgeon. PMID- 8724271 TI - Inoperable liver echinococcosis treated by oral antiparasitic drugs. PMID- 8724273 TI - Flexible bronchoscopy. AB - Fiberoptic bronchoscopy is an excellent tool for the evaluation of the airways and lung parenchyma. The history of flexible bronchoscopy, anesthesia, technique, indications, contraindications, and complications in using the bronchoscopy are reviewed. PMID- 8724274 TI - The documentation of thoracic endoscopy. AB - Thoracic endoscopy encompasses bronchoscopy, esophagoscopy, laryngoscopy, mediastinoscopy, and thoracoscopy (pleuroscopy). In this article, the role of documentation as it applies to bronchoscopy is discussed. However, the discussion is generally applicable to other thoracic endoscopy procedures. PMID- 8724275 TI - Bronchoscopic biopsies and bronchoalveolar lavage. AB - Bronchoscopy with biopsies, bronchoalveolar lavage, and other sampling techniques are frequently needed to establish definitive diagnoses of pulmonary disorders. Combinations of specimens provide superior results to single specimens alone for lung cancer, including those which are endoscopically visible and peripheral in location. Transbronchial biopsy is useful to establish tissue diagnoses in certain diffuse parenchymal lung diseases with specific recognizable histologic patterns such as sarcoidosis or eosinophilic granuloma, but it is less useful for disorders such as interstitial pulmonary fibrosis. Patients with tuberculosis can be diagnosed by performing bronchoscopy, but other sampling techniques are equally good and safer for the bronchoscopist and other health care workers. Bronchoalveolar lavage is especially valuable for confirming infectious complications in immunocompromised hosts, and it also has great potential to elucidate basic mechanisms of pulmonary diseases in research applications. PMID- 8724276 TI - The staging of lung cancer by bronchoscopic transbronchial needle aspiration. AB - The development of flexible transbronchial needles for hilar and mediastinal aspiration had made possible noninvasive staging at the time of the initial diagnostic bronchoscopic examination. Combining the specificity of TBNA with the sensitivity of thoracic CT scanning greatly enhances the utility of this procedure. A bronchoscopically oriented staging system that utilizes the complementary advantages of both of these techniques is outlined. The staging of bronchogenic carcinoma by TBNA is a safe and cost-effective technique that can be routinely utilized to stage lung cancer. Ultimately, its value will depend upon whether the initial encouraging reports of its use can be reproduced by bronchoscopists in practice. PMID- 8724277 TI - Pediatric bronchoscopy. AB - Endoscopic techniques for evaluation and management of airway and pulmonary problems in pediatric patients have matured greatly in the past decade, especially with the introduction of flexible instruments suitable for use in the smallest infants. Careful selection of the most appropriate instrument and technique by well-trained medical or surgical endoscopists will result in safe and effective diagnosis and treatment of a wide variety of conditions affecting the upper and lower respiratory tract. PMID- 8724278 TI - Airway foreign bodies. AB - The removal of airway foreign bodies can be challenging even for the most experienced endoscopist. A familiarity with and working knowledge of older time tested techniques and instrumentation as well as of the newer rigid and flexible fiberoptic equipment is essential for all who desire to accept these challenging situations. Each instrument has inherent advantages, disadvantages, and limitations in certain situations. Occasionally, a foreign body may be removed more safely through open surgical procedures. A knowledge of the lessons learned and techniques developed by pioneering endoscopists coupled with continuing practice of different endoscopy techniques with a variety of instrumentation will prepare the endoscopist to handle unusual foreign body dilemmas with greater skill and safety. PMID- 8724279 TI - Laser bronchoscopy. AB - Laser bronchoscopy allows reestablishment of patent airways in patients with exophytic intraluminal tracheobronchial obstruction from malignancy. Patients with benign strictures can also be treated, thereby avoiding open surgical repair in selected instances. Because of laser-tissue interactions, and because patients often have terminal illness or severe underlying medical problems, these are potentially high-risk procedures. Proper resection techniques, experience, and repetitive performance are necessary to ensure satisfactory patient safety and outcome. PMID- 8724280 TI - Endoscopic brachytherapy. AB - Intraluminal radiation for the local control of bronchogenic carcinoma has recently undergone rapid technological progress. Remote afterloading of high intensity radiation sources into endoscopically placed multiple small catheters with computerized optimization of dosimetry is the state-of-the-art. Fractionated high dose rate (HDR) outpatient treatment has been shown to reduce airway obstruction and improve performance status and quality of life. PMID- 8724281 TI - Expandable stents. AB - Expandable metallic stents are effective in selected patients with malignant or benign airway stenoses. When used for malignant lesions, the primary purpose of the stent is to improve the quality of life; stents are usually chosen for palliation of symptoms in recognition of the low likelihood of success for other therapy. For patients with benign stenoses, the stents provide a permanent source of structural support to alleviate the narrowed segment. The advantages of the expandable metallic stents are as follows: (1) they can be inserted through an endotracheal tube or under local anesthesia with relative simplicity under fluoroscopic guidance; (2) they do not impair the drainage of sputum because ciliary movement is not interrupted; (3) over a period of a few weeks, the meshwork is gradually covered with mucosa as the stent becomes incorporated into the airway wall; (4) ventilation usually is not impaired if the metallic mesh stent covers another nonstenosed bronchus, because the interstices of the stent are nonobstructive; and (5) they are dynamic and continue to expand over time, particularly if concurrent treatment achieves an effect on the lesion that caused stenosis. Disadvantages of the expandable stent include (1) they often are only temporarily effective for tracheobronchial stenosis due to intraluminal tumor or granulation tissue, both of which can grow between the wires; (2) they are considered permanent stents because removal is difficult; and (3) they can be poorly positioned during placement or can become displaced by progressive migration after placement, and they cannot be repositioned. A relative contraindication to insertion is an inflammatory process or infection that can predispose to granulation formation, particularly at the points of maximal contact pressure of the stent to the airway mucosa. In the presence of inflammation, it may be better to use a silicone prosthesis until the inflammatory process subsides and fibrosis occurs. Granulation tissue is less likely to occur in areas of established scar than in areas of acute inflammation. In circumstances in which it is essential that a stent remain only temporarily, an expandable stent should not be inserted in favor of a silicone stent, which can be removed. In the future, expandable stents may have silicone coverings or may be constructed of materials that facilitate removal. Until that time, expandable stents should be considered permanent and nonremovable. Subtle differences exist among the available stents. Standard is low-profile expandable construction from wire mesh. A relatively minor difference is the slightly wider expansibility of the Gianturco stent, a quality that makes it the best suited of the three stents for lesions that involve tracheomalacia. Perhaps the only major difference between the Wallstent and the Gianturco and Palmaz stents is its better ability to conform to tortuous lesions. In an acutely angled stricture, a Wallstent offers a better opportunity for successful placement than other stents. The filamentous meshwork of the Wallstent allows it to bend and conform better to distorted airways. The Gianturco and Palmaz stents have little longitudinal elasticity, which makes them less effective in a tortuous or highly angulated airway stenosis. Expandable stents have demonstrated their efficacy and exposed their limitations in the treatment of airway stenoses. Refinements in design should help to lessen specific disadvantages and problems. Covered expandable stents, a realistic prospect, have specific advantages over standard expandable stents; they will be removable and prevent tumor ingrowth. Current investigative work with such prostheses for the vascular system may provide the foundation for their investigative use in the airway. In essence, two categories of expandable stents are evolving, covered and uncovered, each having unique features adaptable to the specific clinical needs. PMID- 8724282 TI - The use of fiberoptic endoscopy in anesthesia. AB - The fiberoptic bronchoscope has contributed greatly to the practice of anesthesiology over the past 30 years. It has become an indispensable tool in the approach to the difficult airway, as well as the placement and positioning of double-lumen tubes for thoracic surgery. The equipment, preparation, and methods for using the endoscope in anesthesia, are presented here. PMID- 8724283 TI - Bronchoscopy in the intensive care unit. AB - With the prevalence of bronchopulmonary disease in the intensive care unit, bronchoscopy has become an essential tool for the management of patients. This article describes the variety of situations in which bronchoscopy can be of assistance in establishing diagnosis, managing the difficult airway, and supporting the patient's suffering of trauma, hemoptysis, atelectasis, and pneumonia. PMID- 8724284 TI - Future diagnostic procedures. AB - Although the exact proportion of patients with carcinoma in situ in whom disease will progress to invasive lung cancer is not known, and although there have been reports that some individuals may continue to show malignant cells in sputum for several years without symptoms or abnormality on chest radiograph, untreated or suboptimally treated carcinoma in situ has been shown to progress to invasive cancer or metastatic disease. A study by Frost and co-workers showed that cancer developed in approximately 10% of individuals with moderate atypia and 40% of those with severe atypia in sputum cytology. Therefore, the proportion of individuals with carcinoma in situ in whom invasive cancer will develop is likely to be greater than 40%. Lung cancer is almost uniformly fatal when untreated, and, if the disease is allowed to progress to the invasive stage, the results of currently available therapy are poor. A lesson must be learned from cervical cancer screening. As has been shown by Anderson and co-workers, if individuals harboring dysplasia or carcinoma in situ are actively sought for and treated by laser or cryotherapy, the incidence and mortality of invasive cervical cancer can be reduced to extremely low levels. If current work directed toward detecting early lung cancer in sputum cytology specimens in high-risk groups using quantitative image cytometry or molecular markers is successful, the ability to localize small preinvasive lesions with fluorescence bronchoscopy will become even more important for the pulmonologist or thoracic surgeon. Endoscopic ultrasound can be used to determine the depth of tumor infiltration into the bronchial wall or adjacent structures. Biopsy of mediastinal and peribronchial lymph nodes can be performed under sonographic guidance for more accurate staging. By coupling sensitive diagnostic tools to new treatment modalities, such as chemoprevention and various endobronchial therapies, it is hoped that the traditionally poor prognosis for patients with lung cancer can be altered in the near future. PMID- 8724285 TI - Future therapeutic procedures. AB - Management of disorders of the tracheobronchial tree have advanced significantly since the first use of the rigid bronchoscope by Gustav Killian. The addition of the flexible bronchofiberscope by Shigeto Ikeda offered a second generation of new therapeutic opportunities. The future use of thoracic endoscopy will depend upon the application and improvement of technologies that currently exist in addition to those not yet developed. There are three current technologies which are being improved and may be applicable in the future management of diseases of the tracheobronchial tree and these include cryotherapy, electrosurgery, and photodynamic therapy. PMID- 8724286 TI - Intravenous immunoglobulins in Still's disease: still controversial, still unproven. PMID- 8724287 TI - Monitoring radiographic changes in early rheumatoid arthritis. PMID- 8724288 TI - Antiinflammatory drugs in the treatment of Alzheimer's disease. PMID- 8724289 TI - CD4+ T cell inducible immunoregulatory cytokine response in rheumatoid arthritis. AB - OBJECTIVE: Monocytes and CD4+/CD8+ T cells produce immunoregulatory cytokines that participate in the pathogenesis of various immune disorders. We investigated the secretion of Th1-Th2 cell response cytokine production of CD4+/CD8+ T cells from the synovial fluid (SF) and blood of patients with rheumatoid arthritis (RA). METHODS: Blood and SF purified monocytes, CD4+ and CD8+ T cells were stimulated with bacterial lipopolysaccharides or anti-CD3 antibody, and secretion of various cytokines was determined by bioassay or ELISA: RESULTS: Monocytes from SF and blood of patients with RA produced highly elevated levels of interleukin-1 alpha (IL-1 alpha), IL-6, tumor necrosis factor-alpha (TNF-alpha), and granulocyte macrophage colony stimulating factor (GMCSF), the leading mediators of inflammation. However, CD4+ T cells secreted deficient levels of IL-2 and interferon-gamma (IFN-gamma), but higher levels of IL-4 and IL-10, the typical immunoregulatory Th2 cell response cytokines. CD8+ T cells also produce elevated levels of IL-4 and IL-10 but almost normal levels of IFN-gamma in this disease. CONCLUSION: The cytokine produced by monocytes (IL-alpha, IL-6, TNF-alpha, and GMCSF) and by CD4+ T cells Th2 cell responses (IL-4 and IL-10) may exert immunopathologic and immunoregulatory effects in SF and thus mediate some of the clinical manifestations of RA. PMID- 8724290 TI - Insulin-like growth factor I and its binding proteins 3 and 4 are increased in Human inflammatory synovial fluid. AB - OBJECTIVE: To measure insulin-like growth factor I (IGF-I) and its binding proteins (IGFBP) in human synovial fluid (SF), as IGF-I resistance in the chondrocyte was implicated in the pathogenesis of inflammatory arthritis. METHODS: Ten patients with inflammatory arthritis and 7 controls were evaluated. IGF-I and IGFBP-3 were measured by radioimmunoassay while all IGFBP were evaluated by Western ligand blotting. RESULTS: Serum IGF-I did not differ between patients and controls (22.3 +/- 2.9 vs 22.0 +/- 3.0 nmol/l), while significantly higher values were found in inflammatory SF compared with noninflammatory SF (14.7 +/- 1.5 vs 10.2 +/- 1.2 nmol/l; p = 0.05). IGFBP-3 and IGFBP-4 were the most prominent bands found in inflammatory SF by Western ligand blotting. The 24 kDa IGFBP-4 band appeared faintly in 2 noninflammatory effusions and was nearly absent in others. IGFBP-3 levels in sera from the 2 groups did not differ (2.9 +/ 0.3 vs 2.5 +/- 0.3 mg/l), but were significantly higher in inflammatory compared with noninflammatory effusions (1.5 +/- 0.2 vs 0.9 +/- 0.2 mg/l; p = 0.05). CONCLUSION: The pattern of IGFBP in SF differs in inflammatory versus noninflammatory joints. PMID- 8724291 TI - Antigen binding differences between secreted and cell surface expressed rheumatoid factor derived from inflamed synovium. AB - OBJECTIVE: Rheumatoid factor (RF) is the predominant autoantibody in rheumatoid arthritis (RA). but its role in the pathogenesis of RA remains unclear. We hypothesized that surface RF (sRF) expressed on antigen presenting B cells (B APC) might have different binding specificities than secreted RF. METHODS: We examined RF binding in a novel RF antigen capture enzyme linked immunoassay (ACE) that mimicked sRF binding, and compared it with a direct binding enzyme linked immunoassay (DBE) that mimicked secreted RF. RESULTS: Significant differences in binding characteristics by the same rheumatoid synovial cell (RSC) derived monoclonal RF (mRF) were observed between the ACE and the DBE. For example, several mRF that demonstrated the classical Ga binding pattern (binding to IgG1, 2, and 4) in the DBE showed considerable binding to selected IgG3 proteins in the ACE; and several mRF that bound only to rabbit IgG in DBE bound to human IgG in the ACE. CONCLUSION: These RF reactivity differences may be attributed to conformational modifications in the RF and IgG molecules that expose different epitopes or altered binding sites depending on the physical state of the antibody and/or antigen and may be important pathogenically. PMID- 8724292 TI - Incidence, target antigens, and clinical implications of antineutrophil cytoplasmic antibodies in rheumatoid arthritis. AB - OBJECTIVE: To determine the incidence of antineutrophil cytoplasmic antibodies (ANCA) in rheumatoid arthritis (RA), to detect the target antigens of ANCA, and to compare clinical and laboratory data of ANCA+ with ANCA- patients with RA. METHODS: 385 sera of patients with RA were screened for ANCA by indirect immunofluorescence. ANCA+ sera were further analyzed for target antigens by ELISA: The ANCA+ patients were compared to randomly selected ANCA- patients with RA with respect to serological and radiological variables and extraarticular involvement. RESULTS: ANCA were found in 16% of patients with RA (61/385 sera). All sera showed a perinuclear (pANCA) pattern. Antibodies directed against proteinase 3 were not observed. The analysis of ANCA+ and ANCA- patients revealed that the pANCA+ group exhibited significantly higher serological markers of inflammation (p < 0.005) and a higher incidence of rheumatoid factor (p < 0.005). Furthermore, vasculitic involvement was found at a higher frequency (p < 0.05) in the pANCA+ group. Five patients in the pANCA+ group had pulmonary involvement, but none in the pANCA- group. CONCLUSION: pANCA in RA may be a marker for a more aggressive course of disease in respect to serological variables and extraarticular manifestations including rheumatoid vasculitis and lung involvement. PMID- 8724293 TI - Pathogenetic mechanism and prevalence of the stable atlantoaxial subluxation in rheumatoid arthritis. AB - OBJECTIVE: To study the pathogenetic mechanisms responsible for and prevalence of stable atlantoaxial subluxation (AAS). METHODS: Lateral view cervical spine radiographs during flexion and extension were studied in 262 patients with rheumatoid arthritis (RA). RESULTS: AAS was found in 130 patients, 42 (32%) of whom had a stable deformity; in a population based subseries, stable AAS was seen in 16% of the patients with ASS. Atlantoaxial impaction (AAI) was diagnosed in 98 cases and in 37 (88%) of the 42 patients with stable AAS. CONCLUSION: The main cause of the spontaneous stabilization and occasional ankylosis of the atlantoaxial area is the development of erosions and collapse in the lateral facet joints leading to AAI. PMID- 8724294 TI - Determinants of hand function in patients with rheumatoid arthritis. AB - OBJECTIVE: To investigate which clinical variables contribute to the function of the hand in activities of daily living (ADL) in patients with rheumatoid arthritis (RA). METHODS: In 50 patients with RA hand function in ADL was assessed by (1) the dexterity items of the Arthritis Impact Measurement Scales (AIMS), (2) direct observation of the same items by an occupational therapist, and (3) the Jebsen hand function test. A combined hand function factor was constructed by principal component analysis of the 3 hand function measures. Further assessments included measurements of muscle strength, deformity and destruction, range of motion, and local arthritis activity. After bivariate analyses, variables that correlated significantly with the measures of hand function were entered into stepwise multiple regression analyses. RESULTS: The variables having a significant correlation with most of the hand function measures were: pinch and grip strength, patient's assessment of pain and stiffness of the hands, flexion of the thumb and fingers, range of motion of the wrist, alignment of the metacarpophalangeal (MCP) joints, swan neck and Z deformities, and the Larsen erosion score. 78% of the variance of the combined hand function factor could be explained by pinch strength, stiffness of the hands, and the presence of Z deformity and ulnar deviation. CONCLUSION: Pinch and grip strength should be carefully considered in setting goals for conservative or surgical treatment of the rheumatoid hand. In addition, reported stiffness of the hands, malalignment of the MCP joints, and flexion and deformity of the thumb were the most consistent indicators of impaired hand function. PMID- 8724295 TI - Validity and reproducibility of self-administered joint counts. A prospective longitudinal followup study in patients with rheumatoid arthritis. AB - OBJECTIVE: To investigate the reproducibility and validity of self-administered joint counts (JC), measuring tenderness, swelling and the combination of both, in a longitudinal study. METHODS: At the outpatient department a form self administered by patients (SAI-form), was used to measure joint involvement. Concurrent joint examinations were performed by an assessor. The JC and scores for groups of joints by assessors were correlated with those by patients. As a retest the form was completed again by the patients within 10 days. Correlations between the JC measured by the test and retest were computed to investigate reproducibility. RESULTS: Correlations between test and retest were high (> 0.7). Correlations between JC and groups of joints measured by the assessors and by the patients were moderate (0.06). Correlations with other disease activity variables did not differ between assessors' and patients' joint examination scores. CONCLUSION: The patient-administered joint examination was reproducible; however, correlation with the assessors' joint examination was moderate. The value of the self-administered joint count needs further examination and cannot yet replace the assessor's joint examination. PMID- 8724296 TI - Validity of retrospective disease activity assessment in systemic lupus erythematosus. AB - OBJECTIVE: To evaluate the validity of retrospective disease activity assessment derived from clinical charts. METHODS: We prospectively evaluated 37 patients with systemic lupus erythematosus (SLE) in 90 visits using the SLE Disease Activity Index (SLEDAI), the Mexican SLEDAI (Mex-SLEDAI), and the Lupus Activity Criteria Count (LACC) indices. Routine clinical observations were written by rheumatologists blind to index scores. These notes were reviewed 2 years later to obtain retrospective index scores and their validity was assessed using prospective scores as the standard. Statistical analysis was by Spearman's rank correlation coefficient (rs), Wilcoxon matched pairs test, kappa statistic, and intraclass correlation coefficient (ri). We calculated the sensitivity and specificity of retrospective indices to detect active disease. RESULTS: Median retrospective scores were lower in all indices: SLEDAI (4 VS 2, p =0.004, RS = 0.68, ri = 0.30); Mex-SLEDAI (2 vs 1, p < 0.0003, rs = 0.79, ri = 0.31); and LACC (1 vs 1, p = 0.007, rs = 0.65, ri = 0.21). Used to detect active SLE, the retrospective SLEDAI had a sensitivity of 0.68 and a specificity of 0.86; corresponding values for the Mex-SLEDAI were 0.72 and 0.91, and for the LACC, 0.77 and 0.76. CONCLUSION: Retrospective disease activity indices tended to provide lower scores than prospective evaluations. They often missed patients with mildly active disease, but when positive they were good predictors of disease activity. PMID- 8724297 TI - Quinolinic acid in patients with systemic lupus erythematosus and neuropsychiatric manifestations. AB - OBJECTIVE: To evaluate the relationship between quinolinic acid, a neuroactive metabolite of L-tryptophan, and neuropsychiatric manifestations of systemic lupus erythematosus (SLE). METHODS: Forty specimens of cerebrospinal fluid (CSF) were obtained from 39 patients with SLE who were evaluated for 40 episodes of neuropsychiatric dysfunction. The diagnosis of the neuropsychiatric dysfunction was determined clinically. CSF and serum specimens were analyzed for levels of quinolinic acid without knowledge of the clinical diagnosis. RESULTS: Neuropsychiatric dysfunction attributed to SLE (NPSLE) was confirmed in 30 patient-episodes (Group 1), whereas in the other 10 (Group 2) other etiologies were felt to explain their CNS dysfunction. The median levels of CSF quinolinic acid for Group 1 (232.5 nmol/l) were significantly higher than those for Group 2 (median 38.2 nmol/l) (p < 0.014). CSF and serum quinolinic acid levels correlated significantly (p < 0.003) but there was not correlation between CSF quinolinic acid and CSF protein concentrations or white blood cell counts. CONCLUSION: We conclude that elevated quinolinic acid levels in the CSF and serum may be associated with NPSLE and could possibly play a role in its pathogenesis. PMID- 8724298 TI - Dehydroepiandrosterone sulfate is positively correlated with soluble interleukin 2 receptor and soluble intercellular adhesion molecule in systemic lupus erythematosus. AB - OBJECTIVE: To determine whether dehydroepiandrosterone sulfate (DHEAS) is linked with soluble immune mediators in systemic lupus erythematosus (SLE). METHODS: DHEAS and various soluble immune mediators were measured by ELISA in the serum of 35 patients with SLE (26 women, 9 men) and in 41 control subjects. RESULTS: DHEAS was lower in patients with SLE compared to controls (male 1.29 +/- 0.32 vs 3.04 +/- 0.33 micrograms/ml, p < 0.001; female 0.75 +/- 0.12 vs 2.16 +/- 0.18 micrograms/ml, p < 0.001). The DHEAS reduction was in part dependent on prior glucocorticosteroid treatment (p < 0.02). After adjustment for multiple comparisons, there was significant negative correlation between steroid dose and DHEAS (RRank = -0.426, p = 0.005), but with none of the soluble immune mediators. No significant difference in the percentage of steroid treated male and female patients was found (p = 0.220). However, there was positive correlation between DHEAS and soluble interleukin 2 receptor in women, but not in men, with SLE [RRank = 0.747 (n = 26, p < 0.0001) vs RRank = -0.1333( n = 9, p = 0.366)] and between DHEAS and soluble intercellular adhesion molecule in women, but not in men, with SLE [RRank = 0.509 (n = 26, p = 0.005) vs RRank = 0.4833 (n = 9, p = 0.094)]. CONCLUSION: These data demonstrate positive interrelation between DHEAS and soluble immune mediators involved in leukocyte function and leukocyte adhesion only in female patients with SLE. PMID- 8724299 TI - Prostaglandin E2 enhances interleukin 8 (IL-8) and IL-6 but inhibits GMCSF production by IL-1 stimulated human synovial fibroblasts in vitro. AB - OBJECTIVE: To examine in vitro the effect of prostaglandin E2 (PGE2) on synovial cell cytokine production. METHODS: Human synovial fibroblasts were stimulated with PGE2 alone or PGE2 in combination with interleukin 1 alpha (IL-1 alpha) (5 ng/ml) and/or indomethacin (10(6) M) and assessed for the production of IL-8, IL 6, and granulocyte macrophage colony stimulating factor (GMCSF) at the protein and messenger RNA (mRNA) levels. RESULTS: PGE2 alone had little detectable effect on IL-8 or GMCSF; however, a small enhancement of both IL-6 mRNA and protein levels was seen. While all cytokines were markedly stimulated by IL-1 alpha), co addition of the cyclooxygenase inhibitor indomethacin enhanced IL-8 and GMCSF levels, but caused a reduction in IL-6 expression. The addition of PGE2 to cultures stimulated with IL-1 alpha and indomethacin resulted increases in IL-6 mRNA and protein expression while causing a concomitant reduction in GMCSF protein and mRNA expression. PGE2 and illoprost (PGI2 analog) enhanced IL-8 production in stimulated cells. CONCLUSION: While PGE2 alone has limited effects on synovial cell production of IL-8 and GMCSF, its effects are significant in context of IL-1 alpha stimulation; endogenous PGE2 may alter cytokines secreted by mesenchymally derived cells. PGE2 may be an important modulator of cytokine driven inflammation. PMID- 8724300 TI - Evaluation of destructive spondyloarthropathies in hemodialysis by computerized tomographic scan and magnetic resonance imaging. AB - OBJECTIVE: The incidence of destructive spondyloarthropathies (SpA) in hemodialysis can reach 50% after 10 years of treatment. Few studies have analyzed the magnetic resonance (MR) appearance of destructive SpA and data are controversial. Our objective was to evaluate the computerized tomographic (CT) scan and MR appearance of destructive SpA in comparison to infectious spondylodiscitis. METHODS: 23 destructive SpA were observed in 11 patients who had undergone hemodialysis [mean duration of treatment 12 years (89-228 mo)]: 17 SpA of the cervical spine, one of the dorsal, and 5 of the lumbar spine. CT scans at all levels demonstrating narrowing and discovertebral erosions were performed without injection. MRI was performed with T1 and T2* weighted spin echo sequences (0.5T) without gadolinium injection. Radiographs were analyzed by 2 independent examiners. The data concerning MRI of infectious spondylodiscitis were based on a study of 57 personal cases and literature reports. RESULTS: CT scan appearance was similar in all cases, especially showing multiple small well defined lucencies of the vertebral endplates. In 9 patients, MRI showed low signal intensity of the disk and the adjacent vertebral endplates on T1 and T2* sequences. In no case was discal or vertebral high signal intensity on T2* sequence noted. No periodontoidal pseudotumor was observed. CONCLUSION: CT scans and MRI of destructive SpA, even nonspecific, are different from those observed in infectious spondylodiscitis, particularly due to the absence of discal and vertebral hypersignal on T2* sequences. PMID- 8724301 TI - Periarticular ankle sarcoidosis: a variant of Lofgren's syndrome. AB - OBJECTIVE: To focus more attention on the syndrome characterized by periarticular ankle inflammation associated with bilateral hilar adenopathy--some authors regard it as a variant of the Lofgren syndrome, while other suggest it is a distinct clinical entity-- we present a series of 33 cases of periarticular ankle sarcoidosis. METHODS: Sarcoidosis was diagnosed in 330 patients at Bellvitge Hospital over a 20 year period. The medical charts of patients who presented with periarticular ankle inflammation (swelling of both ankles with acute inflammatory signs and preserved articular motion) were reviewed. RESULTS: 33 patients (10%) were identified. Periarticular ankle inflammation began during the spring in more than one-half (54.5%). The mean age was 33 years and patients were predominantly women (66.6%). Most cases (78.8%) were stage I on chest radiograph. All patients had thoracic gallium scans showing increased hilar/mediastinal uptake. Increased characteristic parotid, lacrimal, and/or submandibular uptake was found in 11 of 13 patients who had head scans. Four of 6 whose scans included the legs had bilateral ankle uptake. Erythema nodosum was present concomitantly in 36.3% of patients. No granulomas were found in 4 biopsies of periarticular ankle tissue. All 24 patients who were followed had inactive disease one year after diagnosis. CONCLUSION: The association of periarticular ankle inflammation with bilateral hilar adenopathy is an acute form of sarcoidosis that follows a benign course to total remission. It should be regarded as a variant of the Lofgren syndrome. PMID- 8724302 TI - Amelioration of antigen induced arthritis in rabbits treated with a secreted viral serine proteinase inhibitor. AB - OBJECTIVE: To evaluate the effects of intraarticular administration of SERP-1 protein, a myxoma virus derived antiinflammatory serine protease inhibitor, in an antigen induced arthritis (AIA) model of chronic inflammation. METHODS: AIA was induced in a single joint of 15 rabbits after intraarticular (i.a.) injection of ovalbumin in animals previously immunized to the same agent administered in complete Freund's adjuvant. A 2nd i.a. injection of ovalbumin was given 2 weeks later preceded by 2 daily injections of human transforming growth factor-beta 2. SERP-1 was given by i.a. injection at 2 and 4 weeks after the last i.a. injection of ovalbumin. Three synovial specimens per joint were obtained for synovial histology. Patellar articular cartilage was assessed for collagen integrity and proteoglycan staining. RESULTS: Synovial histology revealed significant diminution in synovial lining cellular hyperplasia, chronic inflammatory infiltration, and cartilage erosion in treated animals, particularly in those receiving 2 i.a. injections of 1 ng of SERP-1. Preservation of articular cartilage was concomitantly noted in treated animals. A dose-response influence on histopathologic change was discernible. CONCLUSION: A viral derived antiinflammatory protein, SERP-1, demonstrates considerable potency in ameliorating chronic inflammation in the AIA model and warrants further investigation as a potential anti-arthritic agent. PMID- 8724304 TI - Relation between flexion angle and intraarticular pressure during active and passive movement of the normal knee. AB - OBJECTIVE: To determine the effect of full active and passive flexion and extension at physiological rates of movement on intraarticular pressure of the normal knee. METHODS: A 22 gauge Intracath catheter was introduced into 7 clinically normal knees and one abnormal knee in 4 subjects. Pressures were recorded via a pressure transducer and correlated with simultaneous recordings of flexion angle from an electronic goniometer while the subject's knees were traverse through active and passive horizontal flexion at a rate of 1 cycle/2 s. RESULTS: The technique produced 6 satisfactory records over mean active and passive flexion ranges of 135 degrees and 148 degrees. On passive movement, pressures remained negative through most of the cycle, rising to main maximum pressures of 10 mm Hg after about 110 degrees of flexion. On active movement, the most common result was a U shaped curve rising from negative in midflexion to positive on full flexion and extension. The mean maximum pressures recorded on active movement were 38 mm Hg on flexion and 18 mm Hg on extension. CONCLUSION: There is no linear correlation between flexion angle and pressure. Under dynamic conditions at zero gravity intraarticular pressure shows a moderate rise on full passive flexion and in most subjects a substantial rise on active flexion and extension. These differences are significant. The factor governing pressure is not the flexion angle but the accompanying soft tissue changes. Failure to use the full movement range could reduce the efficiency of trans-synovial flux. PMID- 8724303 TI - Calcium pyrophosphate dihydrate crystal deposition disease: frequency of tendon calcification about the knee. AB - OBJECTIVE: To delineate the prevalence of tendon and cartilage calcification as well as its possible predilection for age, sex, or side of involvement in calcium pyrophosphate dihydrate (CPPD) crystal deposition disease of the knee. METHODS: We reviewed 225 lateral knee radiographs from 138 patients (M/F: 85/52, 1 unspecified sex, mean age: 73 yrs) to document quadriceps and gastrocnemius tendon calcification. Chondrocalcinosis of the knee was evaluated on 275 knee radiographs (lateral and anteroposterior projection) from 153 patients (M/F: 97/55, 1 unspecified sex, mean age: 73 yrs) to determine the frequency of meniscal and hyaline cartilage calcification. RESULTS: Gastrocnemius tendon calcification was found in 28.4% of knees, which is significantly more common than quadriceps tendon calcification (8.4%). No significant age or laterality difference existed for calcification of either tendon. Quadriceps tendon calcification was more prevalent in men, but no sex difference was present for the gastrocnemius tendon. Calcification of either tendon never occurred without associated chondrocalcinosis. Meniscal calcification was significantly more prevalent than hyaline cartilage calcification; meniscal calcification also was significantly more frequent in men. Increased prevalence of calcification with age was observed in hyaline cartilage but not in the menisci. CONCLUSION: Gastrocnemius tendon calcification is not infrequent in CPPD crystal deposition disease of the knee; identification of such calcification may further delineate this extent of the disorder. PMID- 8724305 TI - The effects of bright light treatment on the symptoms of fibromyalgia. AB - OBJECTIVE: To investigate the effects of bright light treatment on the symptoms of pain, mood, and sleep in patients with fibromyalgia (FM) reporting seasonality of symptoms on the Seasonal Pattern Assessment Questionnaire (SPAQ). METHODS: A randomized 10 week crossover study compared the effects of 4 weeks of "visible electromagnetic fields" (EMF) (light condition; mean 4750 lux, SD 2337 lux) to 4 weeks of "nonvisible EMF" (no light condition) in 14 patients with FM having a minimum SPAQ score of 11. The light visor system (Bio-Brite) was fitted with an opaque filter for the "nonvisible EMF" control condition. RESULTS: No significant differences were found between treatment conditions on tenderness measured with dolorimetry, self-ratings of sleep, pain, mood, and global measures. Mood was not related to pain or sleep. There was significant reduction in depression scores and subjective pain, but increased tenderness and nocturnal awakenings related to time. CONCLUSION: The were no significant differences between the light and no light conditions on pain, mood, or sleep in patients with FM reporting seasonality of symptoms. No relationship was found between mood and the symptoms of FM (i.e., pain, sleep, and fatigue). PMID- 8724306 TI - Assessing functional disability and health status of women with fibromyalgia: validation of a Hebrew version of the Fibromyalgia Impact Questionnaire. AB - OBJECTIVE: To validate a translated version of the Fibromyalgia Impact Questionnaire (FIQ) to be used by Hebrew speaking populations. METHODS: We administered the FIQ to 100 women with fibromyalgia (FM). The FIQ measures physical functioning, work status, depression, anxiety, sleep, pain, stiffness, fatigue, and well being. All patients were asked about the presence and severity (assessed by visual analog scale) of relevant FM symptoms (pain, fatigue, anxiety, etc.); a count of 18 tender points was conducted by thumb palpation, and tenderness thresholds were assessed by dolorimetry. Test-retest reliability was assessed using Spearman correlations. Internal consistency was evaluated with Cronbach's alpha coefficient of reliability. To assess content validity a cutoff criterion of > or = 25% impairment responses was set to indicate a valid item. Construct validity of the FIQ was evaluated by correlating the physical functioning score as well as the separate items with measures of symptom severity, count of tender points, and tenderness thresholds. RESULTS: Test-retest reliability was r = 0.96 for physical functioning, and 0.80-0.96 for other items of FIQ. Internal consistency was alpha = 0.93 at Time 1 and 0.86 at Time 2. Seventeen of 19 items of the FIQ met the > or = 25% criterion. Significant moderate to high correlations were obtained between the FIQ items and severity symptoms, point count, and tenderness threshold. CONCLUSION: The FIQ is a reliable and valid instrument for measuring functional disability and health status in Israeli women with FM. PMID- 8724307 TI - Low prevalences of chronic widespread pain and shoulder disorders among the Pima Indians. AB - OBJECTIVE: To establish the prevalence of shoulder disease and chronic widespread pain in Pima Indians. METHODS: Cross sectional analyses of data from 4230 subjects for shoulder disease and 105 subjects for chronic widespread pain participating in population surveys RESULTS: The prevalence of shoulder disease was 4.4% (95% CI, 3.8-5.1), age-sex adjusted to the 1980 US census population. This is lower than in a study of Caucasians [prevalence ratio (PR) = 0.29, 95% CI, 0.20-0.42 for men and PR = 0.55, 95% CI, 0.41-0.73 for women]. Shoulder disease was associated with non-insulin-dependent diabetes mellitus (PR = 1.67, 95% CI, 1.19-2.36). No chronic widespread pain was identified (95% CI, 0-3.5%). CONCLUSION: Prevalence of these pain syndromes in Pima Indians is lower than in predominantly Caucasian populations. These findings suggest that these populations have different pain perception or different patterns of risk factors for these disorders. PMID- 8724308 TI - Intravenous immunoglobulin therapy in systemic onset juvenile rheumatoid arthritis: a followup study. AB - OBJECTIVE: To report the short and longterm effect of intravenous immunoglobulin (IVIG) in patients with systemic onset juvenile rheumatoid arthritis (SOJRA). METHODS: A retrospective chart review of 27 patients with SOJRA treated with IVIG and followed for 37.1 +/- 18.2 months was undertaken. Indications for treatment were fever, arthritis, or steroid dependency. RESULTS: We treated 27 patients with SOJRA with IVIG monthly for 3-54 months. Six months after IVIG therapy, 20 patients had a least a 50% decrease in at least one of the following: the number of days of fever; prednisone dose; or the number of active joints. Five patients failed to respond to IVIG, and 2 dropped out after 3 and 4 months. At last followup visit (mean 37.6 +/- 18 months), 11 of the initial 20 responder group patients were in remission, while 3 had significantly improved but still had active arthritis, and 6 were now unresponsive. Of the initial 5 patients in the nonresponder group, 4 had nonresponsive arthritis and 1 had improved at last followup. Three patients in the responder group subsequently developed other diseases. CONCLUSION: The main benefit of IVIG therapy to most of our patients was a significant improvement in the systemic features, with resolution of fever and a significant reduction in the steroid dose. The efficacy of IVIG in altering the course of arthritis was less predictable. We suggest that IVIG has a role in the management of SOJRA, but it should be limited to patients with severe SOJRA in whom prolonged unresponsiveness to standard therapy is present. PMID- 8724309 TI - Intravenous immunoglobulin in the treatment of polyarticular juvenile rheumatoid arthritis: a phase I/II study. Pediatric Rheumatology Collaborative Study Group. AB - OBJECTIVE: To obtain preliminary information about the safety and efficacy of intravenous immune globulin (IVIG: Iveegam, Immuno AG, Vienna) in the treatment of polyarticular juvenile rheumatoid arthritis (poly-JRA) resistant to other forms therapy. METHODS: We used a multicentered, phase I/II blinded-withdrawal design with stratified entry. All patients began by receiving open infusions of IVIG at a dose between 1.5 and 2.0 g/kg/infusion (100 g maximum) bimonthly for the first 2 months, then monthly for up to 6 months. Beginning at Month 3, those who met the criteria for "clinically important improvement" were randomized to receive monthly infusions for 4 months of either placebo or IVIG in a double blind (DB) phase. Patients were permitted nonsteroidal antiinflammatory drugs, slow acting antirheumatic drugs, and low dose (< 10 mg/day) prednisone at constant doses. An "early escape" provision in the DB allowed those who showed "clinically important worsening" to again receive IVIG (if taking placebo) or a higher dose of IVIG (if taking the lower dose of IVIG). RESULTS: Efficacy. Twenty five children entered the trial and 19 (76%) met the criteria for "clinically important improvement" during the open phase (OP) and entered the DB. Three patients completed the OP but failed to meet the criteria for response, and 3 patients dropped out of the OP, none of whom showed benefit from IVIG. Treatment effect sizes produced by IVIG were moderate to large for all variables in the OP. Patients who continued IVIG in the DB continued to show improvement over that achieved in the OP. Those given placebo showed a rapid loss of efficacy, suggesting IVIG has a limited duration of effect after discontinuation. Safety. No patient developed serious or unexpected adverse side effects in the open or DB phases, and none dropped out of the study due to toxicity or side effects. CONCLUSION: Substantial clinical improvement from IVIG is produced in about three fourths of patients with poly-JRA during open administration, but the duration of the beneficial effect is short after discontinuation. Those with disease < 3 years' duration may be more likely to respond than those who have had their disease for > 5 years. Short term safety is excellent. PMID- 8724310 TI - Subclass distribution of maternal and neonatal anti-Ro(SSA) and La(SSB) antibodies in congenital heart block. AB - OBJECTIVE: To compare the subclass distribution of anti-48 kDa La(SSB) and anti 52 and 60 kDa Ro(SSA) antibodies in the maternal and neonatal circulation, in pregnancies affected and unaffected by the development of congenital heart block (CHB). METHODS: Sera were obtained from 32 mothers (during 34 pregnancies 23 complicated by CHB and 11 healthy) demonstrated to have anti-Ro(SSA) and/or La(SSB). Maternal and neonatal autoantibodies were evaluated for subclass distribution by ELISA: RESULTS: All 4 subclasses of anti-Ro(SSA) and La(SSB) antibodies cross the placenta and are detectable in sera obtained from the umbilical cord, IgG1 and IgG3 were the major subclasses represented in the 48 kDa La(SSB) and 52 kDa Ro(SSA) responses. All subclasses, including IgG2 and IgG4, were observed in about one-third of the anti-52 kDa Ro(SSA) and 48 kDa La(SSB) responses. In contrast, anti-60 kDa antibodies were, with rare exception, confined to IgG1. Except for anti-48 kDa La(SSB) IgG3 antibodies, no significant differences were observed between affected and unaffected pregnancies in the ratio of maternal to neonatal levels of any of the antibody subclasses. Overall, there were no significant differences in the subclass profiles between mothers whose children had heart block and those who did not. CONCLUSION: The IgG subclasses of anti-48 kDa La(SSB) and anti-52 and 60 kDa Ro(SSA) do not account for the susceptibility of one fetus versus another for the development of CHB. Anti-60 kDa Ro(SSA) antibodies are more restricted in subclass distribution than anti-52 kDA Ro(SSA) or 48 kDa La(SSB) responses. PMID- 8724311 TI - Diffuse interstitial pulmonary amyloidosis in rheumatoid arthritis. AB - We describe a woman with seronegative rheumatoid arthritis (RA) who presented with diffuse septal pulmonary amyloidosis mimicking interstitial rheumatoid lung disease. Her systemic amyloidosis was diagnosed by biopsy of the kidney, stomach mucosa, and salivary glands 28 years after the onset of RA. Diffuse interstitial pulmonary infiltrates had been noted since that diagnosis, but infiltrates had been considered due to rheumatoid lung. Results of pulmonary function tests, which revealed restrictive changes and decreased diffusion capacity, were also compatible with rheumatoid lung disease. Pulmonary amyloidosis was diagnosed at autopsy. Pulmonary amyloidosis should be considered a cause of pulmonary infiltrates in patients with longstanding RA. PMID- 8724312 TI - Steroid sparing effect of methotrexate in relapsing polychondritis. AB - Relapsing polychondritis is a rare inflammatory disorder causing recurrent inflammatory reactions in the cartilaginous structures. It often worsens as prednisone is tapered. We describe 3 biopsy proven patients with relapsing polychondritis in whom methotrexate was useful as a steroid sparing agent in the management of auricular chondritis. PMID- 8724313 TI - Bilateral patellar metastases as the clinical presentation of bronchogenic adenocarcinoma. AB - Common symptoms of the musculoskeletal system may occur as a rare presentation of an underlying malignancy. We describe a case of bronchogenic adenocarcinoma presenting as bilateral knee pain with arthritis due to bilateral metastases to the patellae. We also review the literature of patellar metastases. PMID- 8724314 TI - Proposal for classification criteria for idiopathic arthritides of childhood. PMID- 8724315 TI - Proposal for classification criteria for idiopathic arthritides of childhood. PMID- 8724316 TI - Acute local reactions after intraarticular hylan for osteoarthritis of the knee. PMID- 8724317 TI - How low can you go? Use of very low dosage of gold in patients with mucocutaneous reactions. PMID- 8724318 TI - Acute local reactions after intraarticular hylan for osteoarthritis of the knee. PMID- 8724319 TI - Lupus anticoagulant and thrombosis in relapsing polychondritis. PMID- 8724320 TI - Does male:female sex ratio in ankylosing spondylitis change with age? PMID- 8724321 TI - Concentration-effect studies--making sense of sparse concentration data? PMID- 8724322 TI - IUDs--an update. AB - The intrauterine devices (IUDs) now available offer almost available offer almost complete protection from pregnancy. Some models are effective longer than any other reversible family planning method. Correctly inserted, IUDs are safe for women at low risk of sexually transmitted disease. In fact, because IUDs prevent pregnancy so well, they save many lives. PMID- 8724323 TI - Assisted suicide and euthanasia: what about the clinical issues? AB - This paper aims to address the clinical issues involved in a patient's request for assisted suicide. The psychiatric and broader psychosocial issues for the dying patient, their family and their treating doctor have been largely unaddressed in the debate concerning euthanasia to date. A range of the clinical issues that need to be incorporated in the ethical and legal considerations are reviewed. The reasons for a patient seeking suicide as a treatment are complex and go beyond questions of a patient's right to die. The request for euthanasia needs to be seen in the context of the patient's circumstances, including relationships with and attitudes of carers and health professionals, along with patterns of psychiatric disorder and psychiatric symptoms in the medically ill. The clinical issues involve not only the diagnosis or management of psychiatric disorder but also the acknowledgment of the factors influencing an individual's adjustment to the threat of illness and death. The dynamics of family interactions and doctor-patient relationships in this setting are factors that may impinge upon a request for assistance to die. These factors may be more important than the severity of a person's illness or their quality of life, and are less likely to be recognised and addressed in situations of professional isolation. There are critical issues facing psychiatry in new legislative developments. PMID- 8724324 TI - The present state of psychiatric epidemiology. AB - The seven uses of epidemiology identified by Morris all have direct relevance for mental disorders. But psychiatric epidemiology has a number of distinguishing features: it has to depend on a nosology that may lack validity; it depends largely on subjective symptoms for its data; the morbidity it deals with is really a continuum, and not a dichotomy of cases and unaffected persons; the measurement of morbidity using scales or standardised interviews is open to considerable error; and the unbiased assessment of environmental exposures proves to be technically challenging. Some of these features have been the basis of interesting research developments. The main milestones in the progress of psychiatric epidemiology are reviewed, together with some of the current issues of general interest, including activities in Australasia. PMID- 8724325 TI - Drug strategies and treatment-resistant schizophrenia. AB - OBJECTIVES: The aims of the paper are to review the notion of treatment resistance in schizophrenia and consider the factors important in determining non responsiveness to standard neuroleptic treatment, and to review the strategies currently available in the treatment of such patients, including an evaluation of recently-introduced, novel drug treatments. METHOD: A selective review of the literature relating to treatment resistance was undertaken using medline searches, followed by cross-checking for further articles identified in these references. RESULTS: The various treatment approaches available are considered, including adjunctive treatment with lithium or carbamazepine. The risks and benefits of high dose antipsychotic treatment are discussed. The possible benefits and side-effects of new treatments, particularly the atypical neuroleptics, are also reviewed. CONCLUSIONS: The reasons why a proportion of patients with schizophrenia fail to respond to standard neuroleptic treatment are ill-understood. Nevertheless, initial assessment should include identification of any factors that may be related to a patient's poor response, such as poor compliance, substance use or epilepsy. This may help to determine an appropriate treatment strategy. There is a need to be systematic and to ensure that patients be given an adequate trial of each treatment tested in terms of duration and dosage. The available evidence does not support the use of high doses of neuroleptics for the majority of patients. Adjunctive treatments, such as lithium, carbamazepine or benzodiazepines may be beneficial in non-responsive patients, particularly if certain target symptoms are present. Atypical neuroleptics, particularly clozapine, have proved particularly effective in non responsive patients as well as those sensitive to the motor side-effects of standard drugs. However, the high risk of agranulocytosis with clozapine is a problem; also the drug and the necessary haematological monitoring are expensive. There are hints that some of the other, new, atypical neuroleptics have some benefit in non-responsive patients, but controlled studies are required. PMID- 8724326 TI - Restlessness: the anatomy of a neuropsychiatric symptom. AB - OBJECTIVE: The aim of this paper is to review the literature on restlessness and related syndromes in order to examine the different causes and clinical descriptions, and to present a pathogenetic model that would incorporate its diverse aetiology. METHOD: A literature search was undertaken with restlessness, agitation, akathisia, hyperactivity, fidgetiness and jitteriness as key words. RESULTS: Causes of restlessness are diverse, and its distinction from other descriptions, such as agitation and hyperactivity, is poorly defined in the literature. Detailed descriptions of the syndromes are therefore lacking. The neuroanatomical basis of restlessness may consist of abnormalities in the cortico subcortical neuronal circuits, the complex regulation of which may explain why different causes often lead to a common end result. CONCLUSIONS: The terms used to describe restlessness and related disorders should be standardised, and the clinical manifestations investigated pedantically. Human and animal studies should investigate the pathophysiology so that intervention can be based on the underlying mechanisms. PMID- 8724327 TI - The psychological sequelae of miscarriage: a critical review of the literature. AB - Miscarriage, although a common event in pregnancy, has been frequently overlooked in psychological research. This paper reviews the literature on the psychological sequelae of miscarriage, including the shortcomings of that literature. Best understood against the background of psychological changes in early pregnancy, the literature reveals aspects of grief specific to miscarriage. Important components of this grief comprise high levels of guilt, the loss of part of the self and a large impact upon personal identity. The psychological sequelae impact upon other family members, including partners and surviving children. Psychiatric consequences include depression, anxiety and posttraumatic stress disorder. Given the potentially serious nature of these sequelae, it behooves the psychiatrist to inquire routinely about pregnancy loss in all female patients. PMID- 8724328 TI - Consultation-liaison psychiatry in an obstetric service. AB - OBJECTIVE: The aim of this paper is to provide an overview of the work of a consultation-liaison (C-L) psychiatry service to an obstetric inpatient unit in a university affiliated teaching hospital, with the aid of a comprehensive clinical database. METHOD: The MICRO-CARES clinical database system was used for prospective recording of data on all obstetric inpatients referred to the C-L psychiatry service and the hospital clinical database was used to compare referred inpatients with all obstetric inpatients. RESULTS: Ninety patients were referred in 3 years, a referral rate of 1.2% of obstetric admissions. There was no bias in referral on age, but there were significantly more unmarried patients in the referred group (p < 0.001). Referred patients had a significantly prolonged length of stay: a mean of 9 days for referred patients, with 6 days for all obstetric patients (p < 0.001). The most frequent reasons for referral were: coping problems, depression, anxiety/fear and past history of major psychiatric illness. The most common DMS-III-R psychiatric diagnoses were: Personality Disorders (19%), Mood Disorders (17%), Schizophrenic Disorders (15%) and Adjustment Disorders (13%). At least one confirmed diagnosis was made in 77% of patients, but 42% of diagnoses remained differential. There were significantly more patients with diseases of the nervous system, endocrine and circulatory disorders in the referred group (p < 0.001). Recommendations of psychosocial interventions predominated over psychopharmacological interventions, and concordance with these was 97% and 98%, respectively. Issues in the relationship of obstetrics and psychiatric C-L services are discussed and future directions indicated. CONCLUSIONS: The referral rate was low compared with other specialty units. This is probably due to factors inherent in obstetric practice. A wide spectrum of psychiatric disorders was referred, including a higher than expected number of women with postpartum psychotic disorders, the majority of whom were managed successfully in the obstetric ward. There is a need for increased liaison with obstetric and community services and for collaborative research, with a particular emphasis on the prevention of psychiatric morbidity associated with pregnancy. PMID- 8724329 TI - Psychological morbidity in obstetric-gynaecology patients: testing the need for expanded psychiatry services in obstetric-gynaecology facilities. AB - OBJECTIVE: The present study set out to determine general levels of distress, anxiety and depression in an obstetric-gynaecology inpatient population and to ascertain levels of patient satisfaction with services. METHOD: 200 English speaking consecutive admissions to the hospital filled out a package of questionnaires consisting of a demographic data form, the 30-item General Health Questionnaire, the Speilberger State-Trait Anxiety Inventory, the Inventory to Diagnose Depression and a Patient Satisfaction with Services Questionnaire. 134 packages were fully completed. RESULTS: The most significant findings were: first, that high levels of anxiety prevailed, with approximately 1/3 of patients at or above the 75th percentile for anxiety levels; and second, that those patients born in non-English speaking countries were significantly more dissatisfied with their care. CONCLUSIONS: The results of this study suggest that there is a need for increased awareness of and further research into the psychological needs of obstetric-gynaecology patients. PMID- 8724330 TI - Progress and evaluation of a consultation-liaison psychiatry service to an obstetric-gynaecology hospital. AB - OBJECTIVE: Recently we described the establishment and first 6 months of a psychiatric consultation-liaison service to a Melbourne obstetric-gynaecology teaching hospital. The follow-up report evaluates the service in two ways: first, it compares referral data for the 12 months of 1992 with that of the first 6 months of operation in 1990; and second, it reports on results of a survey of referrer and patient satisfaction with the service. METHOD: Referral data were collected for the data comparison from the consultation-liaison referral book and patients' files. Referrer and patient satisfaction was evaluated by questionnaires sent to 45 medical staff, 7 charge nurses, and 100 consecutive patients. RESULT: A fairly constant referral rate for inpatients has emerged at 0.8%, this very low referral rate being some cause for concern. An internal shift in the type of referrals over time has developed, with an increase in obstetric and a decrease in gynaecology referrals. High referrer satisfaction, with 86% of the doctors finding the consultation(s) "very" or "quite" helpful, and high patient satisfaction, with 83% of respondents having found the consultation(s) "very" or "quite" helpful, was found. CONCLUSIONS: Despite high referrer and consumer satisfaction, and objective evidence of need, this pioneering consultation-liaison service in obstetric-gynaecology continues to be grossly under-utilised. Increasing the referral rate will be one of the greatest challenges in the future of this service. PMID- 8724331 TI - An overview of depression-prone personality traits and the role of interpersonal sensitivity. AB - A number of personality styles have been proposed as vulnerability traits to depression. In this paper methodological problems associated with identifying such traits are discussed. These include state effects contaminating personality scales and the issue of depressive heterogeneity. Potential depression-prone personality traits are then discussed, including obsessionality, neuroticism, dependency and cognitive dysfunction. High interpersonal sensitivity as a risk factor to depression is then discussed. PMID- 8724332 TI - Depressive symptoms in patients with HIV infection. AB - OBJECTIVE: The aim of the paper was to assess the frequency and severity of depressive symptoms amongst HIV positive patients attending an out-patient treatment clinic. METHOD: One hundred HIV positive patients attending the out patient clinic at Fairfield Hospital, Melbourne, completed the Beck Depression Inventory (BDI). RESULTS: Forty-four patients scored 14 or more on the BDI. No significant relationship was demonstrated between BDI score and living situation, relationship status, duration of known seropositivity, current CD4 count, or Centres for Disease Control (CDC) category of illness. Significantly more patients scoring in the depressed range were unable to work because of illness. Forty patients reported suicidal ideation. CONCLUSIONS: Depressive symptoms and suicidal ideation are common amongst HIV positive patients, occurring at comparable or greater rates than those found in a variety of other medically ill populations. PMID- 8724333 TI - Phobic disorders are associated with temperament in alcoholic men. AB - OBJECTIVE: To investigate whether in a sample of men with alcohol dependence those with comorbid phobic disorders had higher persistence scores than those without. METHOD: Ninety-nine men with moderate-severe alcohol dependence recruited from an alcohol treatment programme were interviewed using the Structured Clinical Interview (SCID) and completed a Tridimensional Personality Questionnaire (TPQ). RESULTS: Subjects with a comorbid phobic disorder had significantly higher persistence scores compared to those without. CONCLUSIONS: The temperament dimension, persistence, is associated with phobic disorder. PMID- 8724334 TI - Rural psychiatric services. AB - OBJECTIVE: The objective was to describe and evaluate a community mental health service developed during 1991-1992 in an attempt to meet the mental illness needs of an isolated rural community. The setting was the Grampians health region in Western Victoria: this region has an area of 45,000 square kilometers and a population of 182,000. METHOD: The method involved firstly describing the evolution of the service delivery model. This comprised a team of travelling psychiatrists and community psychiatric nurses which succeeded in providing a combined inpatient and outpatient service which was integrated with general practitioners. Secondly, diagnostic and case load descriptions of patients receiving service were compared for both the inpatient and outpatient settings. RESULTS: The results were that reduced reliance on inpatient beds and increased consumer satisfaction were achieved. CONCLUSION: It was concluded that on initial evaluation of the service it was seen to be meeting its objective of treating the seriously mentally ill in an isolated rural community-based setting. PMID- 8724335 TI - Sexual abuse in therapy: gender issues. AB - OBJECTIVE: That sexual abuse in therapy occurs predominantly with male perpetrators and female patients is a phenomenon that requires analysis in terms of gender relations. Such an analysis is undertaken here from the perspective of feminist psychoanalytic and psychosocial theories. METHOD: Data informing the analysis are derived from assessments of 40 women who experienced sexual abuse in therapy. These women had mostly presented depression, 68% had a history of childhood abuse, and one half were themselves helping professionals. RESULTS: The majority were seriously damaged by the abusive therapy. Offenders were chiefly male (90%) and most were senior, well-qualified therapists of high status: some were charismatic leaders or teachers. Such a group cannot be dismissed as marginal, deviant, or ill-informed; a more systemic analysis is necessary to understand how the professions spawn and sometimes protect offenders. CONCLUSION: It is concluded that the professional culture mirrors fundamental problems of gender relations that inhere in the larger socio-cultural context where they are expressed in various forms of sexual abuse and violence. A cultural change requires better education on issues of power and sexual politics. PMID- 8724336 TI - Crime, psychiatry and the insanity defence: a report on some recent reforms in the United States. AB - There have been substantial developments in mental health law in the United States over the last 10-15 years. Once focal point has been the insanity defence, discussed here. The operational consequences of the legal changes remain to be assessed empirically, but informed speculation is possible. Both a description of the reforms and the assessment of their potential effect are relevant to members of the psychiatric profession in Australia, whether they be forensic specialists or traditional practitioners or researchers. Selective consideration of the American experience, as opposed to contemplating Wholesale transposition, is the appropriate posture for Australian policymakers. PMID- 8724337 TI - Computer aids to treatment in psychiatry. AB - OBJECTIVE: A critical discussion is presented of computer-administered treatments in psychiatry, focusing on the results of outcome studies, the range of approaches under investigation, and advantages and disadvantages of this treatment approach. METHOD: A selective review of controlled outcome studies, comparing computer and human therapist administered treatments in clinical populations was made. RESULTS: The small number of studies reported suggest that computer administered forms are no less effective than human therapists for circumscribed, structured cognitive and behavioural interventions, and are acceptable to patients. CONCLUSIONS: Putative computer-administered treatments will become commonplace. They will require thorough evaluation of benefits, risks and costs to determine their proper role in relations to the alternative treatments available. PMID- 8724338 TI - Adrenal carcinoma and hypertension presenting with catatonic stupor. AB - A case of adrenal carcinoma with severe hypertension referred for treatment of "catatonia" illustrates the need for a high index of suspicion of organic mental disorder in medical patients, even when there is clinical evidence of a psychiatric condition and a history of psychosocial stressors. This case illustrates the need for concurrent medical and psychiatric investigation when there is a problem differentiating medical from psychiatric disturbance. PMID- 8724339 TI - A case of unrelenting pursuit of castration. AB - OBJECTIVE: Explore the limitations of the classification of body dysmorphic disorder. CLINICAL PICTURE: A chinese male relentlessly pursued castration to relieve painful erections which were never substantiated. The testes and corporaspongiosum had been removed. No psychotic symptom was prominent. TREATMENT: Cognitive therapy and psychotropics were tried. OUTCOME: The remaining corpora cavernosa was removed subsequently. CONCLUSION: The distorted somatic perception and secondary cognitive amplification (painful erections) could represent a variant of body dysmorphic disorder involving internal body image; diagnostic classifications should accommodate such a category. PMID- 8724340 TI - Use of interpreters in individual psychotherapy. AB - OBJECTIVE: This paper was written after one of the authors treated a case by individual therapy using an interpreter, as patient and therapist spoke different languages. There is little literature on this subject, and this paper describes our findings and recommendations for using this approach. METHOD: A 15-year-old Chinese, Cantonese-speaking in-patient in Hong Kong was treated with individual psychodynamic psychotherapy by an English-speaking Caucasian psychotherapist. The Chinese interpreter attended each session, and therapy was supervised by a bilingual Chinese supervisor. The alternative was to not carry out any therapy, as there was no other therapist available. RESULTS: The patient was treated for a total of 32 sessions. Issues involving language and culture differences between therapist and patient, issues of therapy in a triadic situation involving group dynamics, and specific therapy difficulties raised by the presence of the interpreter are discussed. CONCLUSION: Therapy was not as effective as hoped, but the patient made some improvements. Finding a suitable interpreter is difficult and their role must be well defined. A bilingual supervisor is also needed to monitor the translation as well as supervising the therapist. Psychotherapy through an interpreter is feasible but not ideal. PMID- 8724341 TI - ECT stimulus dosing techniques. PMID- 8724342 TI - Developmental and stage-dependent expression of melanin-concentrating hormone in mammalian germ cells. AB - Melanin-concentrating hormone (MCH) is a cyclic peptide predominantly expressed in the hypothalamus of mammals. This peptide modulates the stress response and regulates many goal-oriented behaviors in the rat brain. MCH mRNA and peptides generated from the precursor, namely MCH and neuropeptide (N) glutamic acid (E) isoleucine (I) amide (NEI), were also found in rodent peripheral tissues including those in adult testis. In the present study, we first examined the cellular distribution and content of MCH gene products and peptide in the testes of adult rats. Using reverse-transcription polymerase chain reaction with MCH gene primers and in situ hybridization with specific 33P-labeled oligoprobes, we characterized the MCH RNA species. Pro-MCH products were revealed through use of immunoperoxidase detection and an RIA with specific MCH and NEI antisera. Both MCH gene transcripts and MCH peptide were found within germ cells at the periphery of some of the seminiferous tubules of adult rats. We further investigated temporal expression of MCH in 7-microns sections of testes from adult rats and mice. MCH was predominantly found in nuclei of spermatogonia at stages II-IV of spermatogenesis and in nuclei of leptotene and zygotene spermatocytes (at stages IX- XIII). MCH was markedly absent in preleptotene spermatocytes at the stage VII-VIII, and MCH immunoreactivity was no longer detectable as spermatocytes underwent the pachytene step. MCH immunoreactivity was also found in some peritubular cells, mainly at stages II-IV. Finally, we studied MCH expression during puberty in the rat, in sterile mutant mice, and in one adult man. We found predominant staining with MCH antiserum over nuclei of immature germ cells as early as 10 days postpartum in the rat and further confirmed the absence of staining except for that in spermatogonia and early spermatocytes during rat postnatal development. MCH distribution was found to be similar in normal and sterile mutant mice, suggesting that MCH expression is not dependent upon the early steps of spermiogenesis. MCH immunoreactivity was found to be confined to the nuclei of spermatogonia and early spermatocytes in the adult man. Our results indicate that MCH is present predominantly within nuclei of spermatogonia and primary spermatocytes in three mammalian species and that its expression is under strong stage-specific and developmental regulation. This peptide may play a role during stem cell renewal and/or differentiation of early spermatocytes. PMID- 8724343 TI - A differentiation antigen of human large luteal cells in corpora lutea of the menstrual cycle and early pregnancy. AB - To investigate the mechanism(s) of luteal cell differentiation, we raised a monoclonal antibody, HCL-1, against human large luteal cells. The antigen was undetectable in growing and preovulatory follicles by immunohistochemistry. The antigen was initially detected on centrally located luteinizing granulosa cells during CL formation. During the midluteal phase, the antigen was expressed at high levels on large luteal cells. Large luteal cells in the CL during late luteal phase and early pregnancy also expressed high levels of HCL-1 antigen, whereas small luteal cells at any stage of the CL did not. Granulosa cells in some atretic follicles weakly expressed HCL-1 antigen. Immunofluorescence staining of enzymatically dispersed luteal cells from human mature CL revealed that HCL-1 antigen was present on the cell surface of large luteal cells. Human granulosa cells isolated from patients who had undergone in vitro fertilization treatment were cultured for 7 days. Indirect immunofluorescence detected HCL-1 antigen on only a few granulosa cells after culture for 1 day and on almost all granulosa cells after culture for 7 days. Flow cytometry of 7-day-cultured cells showed that the percentages of positivity for HCL-1 antigen as well as the mean fluorescence intensities of granulosa cells cultured with hCG (1 IU/ml) were significantly lower than those of the controls (without treatment) (44.3 +/- 3.2% vs. 62.9 +/- 4.0%. p < 0.01; 60.9 +/- 6.7 vs. 82.1 +/- 7.6, p < 0.05). By contrast, the mean fluorescence intensities of cells cultured with interleukin- 1 alpha (10 ng/ml, 105 +/- 6.3, p < 0.05) and tumor necrosis factor alpha (10 ng/ml, 112 +/- 11.2, p < 0.05) were significantly higher than those of controls. These findings showed that the cell surface antigen detected on human large luteal cells by HCL-1 was differentiation-related, and that large luteal cells in the CL of pregnancy are derived from granulosa cells via large luteal cells in the CL of the menstrual cycle. PMID- 8724344 TI - Effect of testosterone on maturational gonadotropin subunit messenger ribonucleic acid levels in the goldfish pituitary. AB - In this study, we investigated the effects of testosterone and a nonaromatizable androgen, 11 beta-hydroxyandrosterone, on maturational gonadotropin (GtH-II) subunit gene expression in the goldfish pituitary. While testosterone treatment at physiological doses resulted in stimulation of GtH-II-alpha and -beta subunit mRNA production, time-course and dose-response studies performed on sexually immature goldfish of mixed sex, using a wider dose range exceeding physiological levels, demonstrated a biphasic response to in vivo androgen treatment. Time related treatment with testosterone and 11 beta-hydroxyandrosterone (20 micrograms/fish) resulted in an initial inhibition of GtH-II subunit mRNA production (12-24 h) followed by stimulation at 72-96 h. In dose-response studies, treatment for 24 h with testosterone resulted in a significant stimulation at the low physiological doses of 0.2 and 2 micrograms/fish. At the supraphysiological level of 20 micrograms/fish, testosterone treatment resulted in no stimulation or in decreased GtH-II subunit mRNA levels compared to the control values. Similarly, treatment with 11 beta-hydroxyandrosterone resulted in a significant stimulation of GtH-II subunit mRNA levels at low physiological concentrations (0.2 microgram/ fish) and an inhibition, or no stimulation, at higher concentrations (2-20 micrograms/fish). In sexually mature goldfish of mixed sex, the biphasic effect of testosterone was not observed in vivo, and treatment with this steroid resulted in stimulation of GtH-II subunit mRNA production in a dose-related manner. To investigate the direct action of testosterone, studies were carried out using isolated goldfish pituitary fragments from goldfish of mixed sex in vitro. Treatment with testosterone at various concentrations was found to stimulate GtH-II subunit mRNA production in pituitary glands obtained from both sexually immature and sexually mature goldfish. Overall, the present study demonstrates a stimulatory effect of testosterone on GtH-II subunit mRNA levels in goldfish. The observed stimulation of basal GtH-II subunit mRNA production by testosterone occurs, in part, through a direct action at the level of the pituitary in both sexually immature and mature goldfish. PMID- 8724345 TI - Puberty occurring either spontaneously or induced precociously in rhesus monkey (Macaca mulatta) is associated with a marked proliferation of Sertoli cells. AB - The pubertal ontogeny of Sertoli cell proliferation in primates is controversial. In the study of the rhesus monkey presented here, we investigated this issue during normal sexual maturation and during precocious puberty induced experimentally with pulsatile GnRH treatment. Cell number was calculated by standard stereological methods employing volume fraction of either seminiferous tubule or, whenever possible, cell nuclei. In the first experiment, testes were collected from normal monkeys at three developmental phases: neonates (7-18 days old, n = 5); juveniles (approximately 17 mo old, n = 4); and adults (5 yr and older, n = 4). In the second experiment, tissue was obtained from an additional three untreated juvenile monkeys and from six other juveniles of comparable age that had been subjected to premature activation of the pituitary-testicular axis for 5 or 10 wk with pulsatile GnRH (1 microgram/min for 3 min every 3 h). The number of Sertoli cells in adults was 6-fold greater than that in neonatal and juvenile animals. A similar increase in number of this somatic cell type was also observed during experimentally induced puberty. A less dramatic proliferation of Sertoli cells occurred during the neonatal-juvenile transition. Although the increase in Sertoli cell number between the juvenile and adult stages of development was paralleled by a comparable change in Ad and Ap spermatogonia, proliferation of these stem spermatogonia during premature activation of the pituitary-testicular axis was less striking. These results lead to the suggestion that, in primates, the onset of puberty is associated with a rapid and substantial proliferation of Sertoli cells, which is subsequently followed by amplification of stem spermatogonia. They also indicate that while Sertoli cells and stem spermatogonia proliferate during infancy and juvenile development, these earlier changes appear to be more insidious than those at puberty. PMID- 8724346 TI - Failure of spermatogenesis to recover despite the presence of a spermatogonia in the irradiated LBNF1 rat. AB - The dose and time response of LBNF1, rat testis to gamma irradiation was studied with use of single doses from 2.5 Gy to 6.0 Gy. Germ cells were initially depleted as a result of killing the radiosensitive differentiating spermatogonia. Some recovery of spermatogenesis was observed at 4 and 6 wk after irradiation as indicated by the repopulation of tubules with germ cells derived from surviving stem spermatogonia. Although spermatogenesis showed additional recovery and was maintained throughout the 60-wk follow-up period after 2.5 Gy, at doses from 3.5 Gy to 6.0 Gy, repopulation indices declined after 6 wk to less than 2%. The numbers of Sertoli cells per nonrepopulating tubule were constant, independent of radiation dose or time. In addition, the nonrepopulating tubules contained an average of one A spermatogonium per 100 Sertoli cells. The size and shape of these cells corresponded to undifferentiated A spermatogonia in nonirradiated control tests. Despite high labeling (40%) and mitotic (20%) indices, the numbers of A spermatogonia changed very little with time, and no differentiated cells were produced in these tubules. The failure of spermatogenesis to recover was not due to hormone deficiency: serum gonadotropin levels increased after irradiation, and serum testosterone remained at control levels. The irradiated LBNF1 rat model may be useful for studying the regulation of differentiation of A spermatogonia. PMID- 8724347 TI - Alterations of intraembryonic metabolites in preimplantation mouse embryos exposed to elevated concentrations of glucose: a metabolic explanation for the developmental retardation seen in preimplantation embryos from diabetic animals. AB - Preimplantation mouse embryos exposed to hyperglycemia, whether in vivo or in vitro, experience delayed development from the 2-cell to blastocyst stage. By comparing metabolites from embryos exposed to high vs. normal glucose conditions, a metabolic explanation for the delayed growth pattern was sought. Fertilized 1 cell embryos obtained from superovulated B5 x CBA F1 mice were cultured for 96 h in medium containing 2.8 mM glucose (C) or in medium with added glucose to give 10 mM, 30 mM, or 52 mM glucose (HG). After incubation, each embryo was quick frozen and freeze-dried. Metabolites were assayed by the ultramicrofluorometric technique and enzymatic cycling to obtain measurable levels in single embryos. Embryos cultured in HG exhibited 7-fold higher intracellular glucose levels than those cultured in C (C: 2.25 +/- 0.6 vs. HG: 16.61 +/- 2.4 mmol/kg wet weight; p < 0.001; C, n = 9; HG, n = 16). This accumulation of glucose was dose-related and stage-dependent. Citrate (C: 1.07 +/- 0.14 vs. HG: 1.98 +/- 0.12; p < 0.001), sorbitol (C: 0.41 +/- 0.06 vs. HG: 0.57 +/- 0.03; p < 0.01), malate (C: 0.81 +/- 0.13 vs. HG: 1.72 +/- 0.17; p < 0.001), and fructose (C: 2.1 +/- 0.3 vs. HG: 5.3 +/- 0.6; p < 0.001) were all significantly higher in HG. Also, these metabolites were highest in the most delayed embryos. Glycogen and 6-phosphogluconate levels were not significantly different. In conclusion, intraembryonic levels of glucose, and polyol pathway and Krebs cycle metabolites are elevated and correspond to the degree of developmental delay. These findings suggest that a metabolic abnormality may be responsible for retarded development experienced by embryos exposed to high glucose. PMID- 8724348 TI - Recombinant soluble human complement receptor type 1 inhibits antisperm antibody- and neutrophil-mediated injury to human sperm. AB - The pathogenesis of antisperm antibody (ASA)-mediated infertility is postulated to be related in part to complement (C)-dependent sperm dysfunction in the female genital tract. We have previously demonstrated that C can be involved in ASA mediated sperm injury by the deposition of activated C3 fragments and the assembly of terminal membrane attack complex (C5b-9) leading to C3-mediated sperm binding to neutrophils or C5b-9-mediated sperm motility loss. This study evaluated the protective effect of recombinant soluble C receptor type 1 (sCR1) on ASA-and C-mediated neutrophil/sperm interaction, neutrophil aggregation, and sperm motility loss. Motile sperm with or without neutrophils were incubated in the presence of 10% C-fixing ASA+ serum or ASA- control sera in the presence or absence of sCR1. After defined incubation periods, the following neutrophil and sperm parameters were evaluated: 1) neutrophil aggregation (by the flow cytometric pulse processing method), 2) sperm phagocytosis (by light microscopy), 3) the deposition of C3 cleavage fragments (C3b, iC3b, and C3d) on motile sperm (by immunofluorescence flow cytometry), and 4) the relation between sperm motility loss and sperm-bound C3d. Only the coincubation of neutrophils with sperm in the presence of C-fixing ASA+ sera resulted in marked neutrophil aggregation (20.5 +/- 0.26% vs. 2.4% +/- 1.6; p < 0.0001) and a concomitant increase in neutrophils containing ingested sperm (71 +/- 5.8% vs. 3.5%; p < 0.0001). Soluble CR1 inhibited ASA- and C-mediated neutrophil aggregation by 46% and sperm phagocytosis by 57%. Motile sperm incubated with C-fixing ASA- sera showed a time-dependent increase in the binding of C3 fragments as detected by flow cytometry using anti-iC3b neoantigen, anti-C3c, and anti-C3d monoclonal antibodies (mAbs). A negative correlation (r2 = -0.930; p < 0.001) was found between the increase in sperm-associated C3d fluorescence and the percentage motile sperm in the presence of ASA- sera. Soluble CR1 (200 micrograms/ml) maximally inhibited the binding of anti-C3b, anti-C3c, and anti-C3d mAbs to sperm by 96%, 83%, and 72%, respectively. Thus, sCR1 abrogated the binding of C3 fragments to human sperm and fully protected sperm from C5b-9-mediated sperm immobilization. These findings suggested the therapeutic potential of sCR1 as an intravaginal pharmacophore to prevent C-dependent sperm dysfunction and related inflammatory events in the female genital tract. PMID- 8724349 TI - Inhibin and activin have antagonistic paracrine effects on gonadal steroidogenesis during the development of the chicken embryo. AB - Primary cultures of gonadal cells from chicken embryos were used to explore the paracrine role of inhibin and activin during gonadal development. Fetal testicular and ovarian cells secreted high amounts of immunoactive inhibin. FSH caused a dose-related increase of cAMP and immunoactive inhibin concentrations in testicular cell cultures. Postreceptor signalling through the protein kinase A (PKA) pathway was confirmed by additional experiments with 8-bromo-cAMP, 3 isobutyl-1-methyl-xanthine (MIX), prostaglandins, forskolin, and choleratoxin. The relative ability of these agonists to stimulate cAMP production did not always correlate with their ability to stimulate inhibin secretion. Experiments with phorbol 12-myristate 13-acetate suggested that the regulation of immunoactive inhibin secretion also involves the protein kinase C (PKC) pathway. In addition, it was shown that recombinant human (rh)-inhibin increases the conversion of pregnenolone to androgens whereas rh-activin has the opposite effect. Recombinant human follistatin, an activin-binding protein, antagonized the actions of rh-activin and to a lesser extent those of rh-inhibin. In conclusion, these results show that during the development of the chicken embryo, gonadal inhibin secretion may be regulated by hormones and by local factors such as prostaglandins. Cross talk between the PKA and PKC pathways may be involved in this regulation. Recombinant human inhibin and rh-activin may have antagonistic roles in the paracrine control of gonadal steroidogenesis. PMID- 8724350 TI - Histone-binding domains in a human nuclear autoantigenic sperm protein. AB - In one of our previous studies, the deduced amino acid sequence of the human nuclear autoantigenic sperm protein (hNASP) revealed two conserved histone binding domains when compared to the Xenopus N1/N2 protein sequence. These histone-binding domains of Xenopus N1/N2 are known to be functional; however, their function in hNASP is unknown. In this study we have determined the number, location, and activity of the histone-binding domains on the primary sequence of hNASP. Purified recombinant polypeptides expressing the full-length hNASP and various deletion constructs covering the entire length of the hNASP sequence were tested by Western blotting and in ELISA for binding to biotin-labeled histones. A positive reaction was detected for the full-length recombinant protein and for the polypeptides spanning the N-terminal region (amino acids [aa] 32-192), and two additional regions: aa 193-352 and aa 353-572. The lack of binding to the expressed C-terminal (aa 573-787), which also contains polyacidic amino acids, suggests that the binding of hNASP to the somatic core histones is a sequence specific as well as an electrostatic interaction. The removal of flanking sequences from the binding domains did not abrogate their ability to bind histones. We conclude that there are at least three functional histone-binding domains in hNASP, two of them encompassing the predicted histone binding sites homologous to the N1/N2 protein, and a third novel domain. Therefore, hNASP may be defined as a nuclear histone-binding protein found in human testis. PMID- 8724351 TI - Occurrence of deoxyribonucleic acid fragmentation during prolactin-induced structural luteolysis in cycling rats. AB - We determined whether fragmentation of genomic DNA, one of the hallmarks of apoptosis, occurs during structural luteolysis in cycling rats. Corpora lutea (CL) were collected from rats at each estrous cycle stage (1800 h), and fragmented DNA was extracted. Only CL from rats at the proestrous stage showed distinct DNA fragmentation. To determine the period of occurrence of DNA fragmentation, CL were collected at several points between 1200 h on the day of proestrus and 0600 h on the day of estrus. Distinct DNA fragmentation was observed from 1800 h (proestrus) to 2400 h (proestrus), and the extent was significantly lower at 0600 h (estrus). It is known that prolactin (PRL) induces structural luteolysis in rats. To examine the role of PRL in luteal DNA fragmentation, 2-bromo-alpha-ergocryptine (BE) was used to suppress the PRL surge on the day of proestrus. CL collected at 1800 h from BE-treated rats did not show distinct DNA fragmentation, and PRL injection offset the effect of BE. Histochemical analysis with a 3'-end labeling technique confirmed the occurrence of DNA fragmentation in luteal tissue. These results suggest that apoptotic cell death occurs during PRL-induced structural luteolysis. PMID- 8724352 TI - Use of synthetic canine relaxin to develop a rapid homologous radioimmunoassay. AB - Canine relaxin (cRlx) was synthesized by a combination of solid-phase methods and sequential site-directed disulfide bond formation. Proof that the intended molecule had been synthesized was obtained by analytical HPLC of the intact and reduced molecule, by amino acid and sequence analysis, and by receptor binding and in vivo mouse interpubic ligament bioassays. Antisera to synthetic cRlx were raised in six male rabbits; these cross-reacted with relaxins of other species, but not with insulin, LH, FSH, hCG, or prolactin (PRL). Three of the antisera neutralized relaxin-induced interpubic ligament formation in estrogen-primed mice in vivo. A new homologous cRlx RIA was developed through the use of rabbit antiserum 79888, synthetic cRlx for standards and 125l-labeled trace, and a goat anti-rabbit lgG-polyethylene glycol precipitant. The new RIA can be completed in 26 h and has a sensitivity of 0.195-0.39 ng cRlx/tube. Intra- and interassay coefficients of variation were 3% and 12.5%. During pregnancy in bitches, serum cRlx rose to about 10 micrograms/ml. Immunoactive cRlx was also detected in serum, colostrum, and milk of lactating bitches, but not in large volumes (100 300 microliters) of serum of pseudopregnant or estrous bitches. Immunoreactive cRlx was also found in seminal plasma, but not in serum, of male dogs. The new homologous cRlx RIA is simple, rapid, sensitive, and specific, and will be used in future studies of canine relaxin physiology. PMID- 8724353 TI - Unusual abundance of arginine and ornithine in porcine allantoic fluid. AB - The concentrations of free arginine, ornithine, and glutamine in porcine allantoic and amniotic fluids were determined on Days 30,35, 40, and 45 of gestation. Arginine and ornithine were the most abundant amino acids in allantoic fluid on Days 35-40 and 45 of gestation, respectively. Arginine and ornithine nitrogen accounted for 40%, 50%, and 55% of the total free alpha-amino acid nitrogen in allantoic fluid on Days 35, 40, and 45 of gestation, respectively. Glutamine was the most abundant amino acid in amniotic fluid during early gestation and was also abundant in allantoic fluid. On Day 45 of gestation, glutamine nitrogen accounted for 41% of the total free alpha-amino acid nitrogen in amniotic fluid. The unusual abundance of arginine (2.5-4.1 mM) and ornithine (1.08-2.52 mM) in allantoic fluid on Days 35-40 of gestation has not been reported for any other biological fluid. These results are novel and interesting with respect to the role of these two basic amino acids in fetal-placental nutrition and metabolism. PMID- 8724354 TI - Ovary mediates the effects of RU486 given during proestrus on the diestrous secretion of luteinizing hormone in the rat. AB - The aim of these experiments was to study the action of proestrous afternoon follicular progesterone secretion on the preovulatory secretion of gonadotropins in the rat. Four-day-cycling rats were given 4 mg of the antiprogestagen RU486 in the morning of proestrus (Day 1), and its effects on the pituitary function during diestrus were compared with those of RU486 given in the morning of estrus (Day 2). The pituitary function was assessed by measuring basal secretion of LH and FSH as well as the pituitary response to either estradiol benzoate (EB) (3 mug/100 g BW at 1300 h on Day 3) or LHRH (100 ng/rat at 1200 h on Day 4). In all experiments, trunk blood was taken at 1300 h on Day 4 to measure serum gonadotropin concentrations. In rats receiving an injection of RU486 on estrus, the absence of only the diestrous progesterone actions increased basal serum concentrations of LH and decreased those of FSH, and as in vehicle-treated controls, EB inhibited and LHRH stimulated LH secretion. In contrast, the absence of both proestrous afternoon and diestrous progesterone actions (as characterized rats treated with RU486 on proestrus) antagonized the inhibitory effect of EB and sensitized the pituitary to LHRH. These effects of RU486 on proestrus are ovary dependent and eliminated by ovariectomy on metestrus. The increased ovarian secretion of testosterone and estradiol-17 beta during diestrus does not mediate the effects of proestrus-administered RU486 on pituitary function: no differences were found in the serum concentrations of estradiol-17 beta in diestrus between the groups of rats treated with RU486, and administration of the antiandrogen flutamide (2 mg/rat at 0900 h on Days 2 and 3) did not reverse the effects of RU486 on proestrus. In conclusion, the results suggest that in the absence of proestrous afternoon progesterone action, the ovaries of the 4-day-cyclic rat keep the pituitary gland in a state of low sensitivity to the inhibitory effects of estradiol and high sensitivity to the stimulatory effects of LHRH. Moreover, the results suggest that the putative ovarian factors involved are factors other than progesterone, androgens, or estradiol-17 beta. PMID- 8724355 TI - Localization of three genes in the hook-shaped hamster sperm nucleus by fluorescent in situ hybridization. AB - We mapped the positions of three different genes in the flat, hook-shaped hamster sperm nucleus to determine the specificity of sperm DNA positioning. The positions of the 5S rRNA gene cluster, the CAD gene, and the class I 1.6 gene were determined by fluorescent in situ hybridization (FISH) in over 50 hamster sperm nuclei for each gene. We first demonstrated by FISH with mitotic chromosomes that the latter two genes were localized on the same chromosome. Within the sperm nuclei, we found that the precise position was variable for each of the three genes, but that there were two areas of preferred localization that contained 26-31% of the nuclear area and within which 80% of the signals were located. Nuclei were then hybridized to two genes simultaneously, using either two genes located on the same chromosome or two genes located on different chromosomes. We found no preference for orientation of one gene relative to the other for either pair of genes examined. This suggested that the relative arrangements of chromosomes within the sperm nucleus are flexible. These data demonstrate that the topographical arrangements of genes within the hamster sperm nucleus have a limited plasticity allowing for a relatively large range of possible localization. PMID- 8724356 TI - Gap junctional protein connexin 43 in bovine corpora lutea throughout the estrous cycle. AB - The present study examined the pattern of expression of the gap junctional protein connexin 43 (Cx43) in bovine corpora lutea (CL) during growth, differentiation, and regression. CL from the early (n = 6), mid- (n = 6), and late (n = 6) luteal phases of the estrous cycle were weighed and divided into several portions. One portion of each CL was frozen in liquid nitrogen for evaluation of protein, DNA, progesterone, and presence of Cx43 by Western immunoblot analysis; another portion was frozen in liquid propane for immunofluorescent staining of Cx43. An additional portion of each CL was dispersed, and the luteal cells were cultured for 2 days, fixed, and used for immunofluorescent staining of Cx43. Weights and DNA, protein, and progesterone contents of CL increased (p < 0.05) from the early to mid-luteal phases and then decreased (p < 0.05) from the mid- to late luteal phases. The ratio of protein to DNA was similar in the early and mid-luteal phases and then decreased (p < 0.05) to the late luteal phase. Western immunoblot analysis revealed bands at 43 kDa that differed in volume (evaluated by densitometry); the early luteal phase volume was greater (p < 0.05) than that at the mid-luteal phase, which was greater (p < 0.05) than that at the late luteal phase. Immunofluorescent staining demonstrated that Cx43 was present in luteal tissues and cultured luteal cells throughout the estrous cycle, and the area of positive staining decreased (p < 0.05) as the estrous cycle progressed. Staining for Cx43 was punctate and localized to the cellular borders. Thus, levels of Cx43 in bovine CL are greatest early in the estrous cycle and are least late in the estrous cycle. These data demonstrate that gap junctions may be important for regulation of luteal growth, differentiation, and regression in the cow. PMID- 8724357 TI - Malignant potential, major histocompatibility complex antigen expression, and genomic imprinting of rat trophoblast cell lines. AB - Invasive growth, variation in major histocompatibility complex antigen expression, and genomic imprinting are important properties of both trophoblast cells and malignant tumors. This study, undertaken to address these three issues, used cultured trophoblast cell lines derived from Day 11/12 rat placentas of all mating combinations of the DA and WF inbred strains. In addition, genomic imprinting was also examined in intact rat placentas from Days 11-19. There was no correlation in trophoblast cells between class I antigen expression, DNA content, and cell ploidy on the one hand and oncogenic potential on the other hand. The constitutive suppression of class II antigens in the trophoblast cells could not be abrogated by treatment with interferon-gamma, whereas such treatment always maximally induced class I antigen expression regardless of the initial resting levels. The trophoblast cells at Day 11/12 expressed both maternal and paternal class I antigens, and studies in whole placental tissues showed that the imprinting of the maternal class I antigens was manifested by a decreased level of expression rather than an absence of expression. Thus, genomic imprinting in the rat placenta is a quantitative, rather than an all-or-none, phenomenon. PMID- 8724358 TI - Regulated assembly of connexin33 and connexin43 into rat Sertoli cell gap junctions. AB - Immunofluorescence and immunogold electron microscopy were employed to examine the assembly of connexins (Cx) 33, 37, and 43 into testis cell gap junctions in mature and postnatal rats. Cx37 was localized by immunofluorescence to the endothelia of blood vessels in both mature and immature testes and was not further characterized. Only Cx43 assembled into Leydig cell gap junctions, but Cx43 also co-assembled with Cx33 in some Sertoli-Sertoli gap junction plaques within and near Sertoli occluding junctions and on adluminal surfaces Assembly of Sertoli gap junctions appeared to be regulated according to the stage of the seminiferous epithelium since Cx33 (and Cx43) immunoreactivity was strong in Sertoli cells from stages II-VII but weak in stages IX-XIV. During postnatal maturation, assembly of Cx33 into gap junctions was regulated independently of Cx43 assembly. Cx43 was present on Sertoli cells of all tubules from postnatal Day 5 through Day 28. In contrast, Cx33 was not apparent on Sertoli cell surfaces until Day 15 and gradually accumulated in all tubules through Day 28. Between postnatal Days 38 and 43, the immunoreactivities of Cx33 and Cx43 became weak in Sertoli cells containing step 9-14 elongated spermatids. Thus, connexin abundance and gap junction composition in Sertoli cells is regulated during testis maturation and the cycle of the seminiferous epithelium. PMID- 8724359 TI - Interferon-gamma inhibits the synthesis and release of renin from human decidual cells. AB - Experiments were performed to examine the effect of interferon-gamma (IFN gamma) on the expression of renin by human uterine decidual cells and decidual macrophages. Exposure of a mixed population of decidual cells consisting of 80% decidualized stromal cells and 20% macrophages to IFN gamma for 4 days caused a dose-dependent inhibition of renin release beginning 2 days after exposure. Renin release on Day 4 was inhibited by a maximum of 83.9%, and the half-maximal effective dose of IFN gamma was 5 ng/ml (290 pM). The inhibition of renin release in response to IFN gamma was accompanied by a comparable inhibition of renin mRNA levels. In addition to inhibiting basal renin expression, IFN gamma potentiated the inhibitory effect of tumor necrosis factor alpha (TNF alpha) on renin expression. IFN gamma also inhibited basal renin release and potentiated the inhibitory effect of TNF alpha by highly purified populations of decidual stromal cells and decidual macrophages prepared by immunomagnetic separation with beads coupled to an anti-human leukocyte antigen (HLA-DR) antibody that binds macrophages but not stromal cells. Reverse transcription-polymerase chain reaction analysis showed that HLA-DR(+) cells express IFN gamma mRNA, and that both HLA-DR(+) and HLA-DR(-) cells express IFN gamma receptors. Since IFN gamma is expressed only by decidual macrophages, the results of this study strongly suggest that IFN gamma inhibits the expression of decidual renin by a paracrine action. PMID- 8724360 TI - Inhibitory effect of peritoneal macrophages on progesterone release from cocultured rat granulosa cells is reversed by dexamethasone: evidence for an action independent of nitric oxide and distal to cyclic adenosine 3',5' monophosphate generation. AB - We investigated the effects of cocultured macrophages on progesterone release from granulosa cells obtained at different stages of the rat estrous cycle and determined whether the immunosuppressive actions of the synthetic glucocorticoid can modulate the effects of cocultured macrophages. Basal and hCG-stimulated progesterone release from granulosa cell-only cultures varied according to the stage of the estrous cycle at which granulosa cells were obtained, but macrophages exerted profound inhibitory effects on both basal and hCG-stimulated steroidogenesis at all stages. These inhibitory effects were partially reversed by the addition of dexamethasone (5 x 10(-7) to 5 x 10(-5) M) to the culture medium. Measurement of nitrite accumulation in the media, as an index of nitric oxide production, showed that granulosa cells produce very low levels of this metabolite compared with macrophages, although granulosa/luteal cells from freshly ruptured follicles of estrous rats produced significantly higher concentrations of nitrite compared with granulosa cells obtained from diestrous and proestrous rats. Dexamethasone had only limited effects in suppressing the accumulation of nitrite, showing that the macrophages were synthesizing nitric oxide synthase and suggesting that the action of dexamethasone was independent of nitric oxide. Macrophages also inhibited the steroidogenic response of granulosa cells to forskolin (5 x 10(-6) M to 10(-4) M) and dibutyryl cAMP (1mM and 0.5 mM) and the potentiation of the response by the addition of hCG. However, this inhibition was reversed by the presence of 5 x 10(-6) M dexamethasone, and responses were comparable with those measured from granulosa cell-only cultures. Overall, the results suggest that macrophage inhibition of progesterone synthesis acts at a site distal to the generation of cAMP and that dexamethasone reverses this inhibition. PMID- 8724361 TI - Expression of heat shock protein 25,000 in rat uterus during pregnancy and pseudopregnancy. AB - In previous studies, we found that the human estrogen-regulated heat shock protein (hsp) 27 (human homologue of rat hsp25) is modulated in the endometrium during the different phases of the menstrual cycle and that it is present in endometrial predecidual cells and in decidual cells attached to the placenta. In the present report, we describe the cell type-specific pattern of hsp25 expression in the rat uterus during the periimplantation period as well as during early and late decidualization and placentation. The hsp25 expression pattern was also analyzed in pseudopregnant rats with deciduomas. Immunocytochemistry was performed with an antibody generated against a chimeric hybrid protein containing the N-terminal of the murine hsp25 and the C-terminal of the human hsp27. During pregnancy at the time of implantation, hsp25 was expressed in the endothelial cells of the endometrial vessels and in the luminal epithelium of the antimesometrial region. As pregnancy advanced, hsp25 appeared in predecidual/decidual cells close to the implantation region and then expanded to the mesometrial region. This expression pattern was very similar during pseudopregnancy. Hsp25 was strongly expressed in trophoblastic giant cells beginning on Day 11 of gestation; less expression was noted in the junctional and labyrinth zones of the chorioallantoic placenta (in some cells lining the vascular spaces). In all the disparate cell types that expressed hsp25, the presence of the protein did not correlate with cell proliferation or with apoptosis but with the state of differentiation. Some placental PRL-family members with molecular weights similar to that of hsp25 are also present in antimesometrial decidua and in differentiated trophoblast giant cells; therefore, in this study we eliminated the possibility that our antibody was recognizing prolactin. We also determined that the hybrid hsp25/27 protein did not bind prolactin receptors, and noted that the hsp25 immunostaining pattern was not identical to that of decidual prolactin. In conclusion, the striking cell type specific timing of hsp25 expression points to hsp25 as a molecule that is important during the implantation, decidualization, and placentation processes. PMID- 8724362 TI - Duration of estrogen exposure prior to follicle-stimulating hormone stimulation is critical to granulosa cell growth and differentiation in rats. AB - Estrogens have been reported to exert both stimulatory and inhibitory effects on granulosa cell function. Previous studies from our laboratory showed that 12 h after administration of diethylstilbestrol (DES; a synthetic estrogen), FSH stimulated granulosa cell proliferation and aromatase activity were increased; however, 48 h after DES, FSH stimulation of both parameters was inhibited. In other experiments, exposure of rats to DES for a period of 26 h blocked ovulation in response to eCG and hCG administration, whereas the same treatment regimen without DES caused ovulation in all treated rats. Thus, DES may in some cases actually interfere with maturation and development of ovulatory follicles. The present study was designed 1) to confirm that the duration of estrogen pre exposure determines the way granulosa cells respond to FSH and 2) to investigate the underlying mechanisms involved. While DES was used in preliminary experiments, the majority of the studies were conducted with estradiol, a natural estrogen, in order to conform as closely as possible to the normal physiology. In the experimental protocol, immature female rats received injections of DES or implants of estradiol pellets 12 h (short exposure) or 36 h (long exposure) before 36 h of FSH treatment. Rats were killed, ovaries removed, and granulosa cells collected at the end of the FSH treatment period. The results demonstrate that exposure to either of these estrogens for 12 h allowed the subsequent FSH stimulation to produce high cellular proliferation, high aromatase enzyme activity, and large amounts of FSH receptor and aromatase mRNA. Estrogen exposure for 36 h, however, resulted in significantly decreased FSH stimulation of all these parameters. These findings confirm that short exposure to estrogen enhances the response of granulosa cells to FSH while longer exposure makes granulosa cells refractory to FSH. This differential sensitivity of granulosa cells to estrogen exposure could help explain how dominant follicles survive to ovulate while others are lost to atresia during ovarian cycles. PMID- 8724363 TI - Structural relationship of sperm soluble hyaluronidase to the sperm membrane protein PH-20. AB - The sperm plasma membrane protein PH-20 has a hyaluronidase activity that enables acrosome-intact sperm to pass through the cumulus cell layer of the egg. In this study we analyzed the relationship of guinea pig PH-20 and the "classical" soluble hyaluronidase released at the time of the acrosome reaction of guinea pig sperm. PH-20 is a membrane protein, anchored in the plasma and inner acrosomal membranes by a glycosyl phosphatidyl inositol anchor. Several types of experiments indicate a structural relationship of PH-20 and the soluble hyaluronidase released during the acrosome reaction. First, an antiserum raised against purified PH-20 is positive in an immunoblot of the soluble protein fraction released during the acrosome reaction. In the released, soluble protein fraction, the anti-PH-20 antiserum recognizes a protein of approximately 64 kDa, i.e., identical in molecular mass to PH-20 (approximately 64 kDa). Second, the enzymatic activity of the released hyaluronidase is completely inhibited (100%) by the anti-PH-20 antiserum. Third, almost all (97%) of the soluble hyaluronidase is removed from the released protein fraction by a single pass through an affinity column made with an anti-PH-20 monoclonal antibody. These findings suggest that the released, soluble hyaluronidase is a soluble form of PH-20 (sPH 20). During the acrosome reaction, PH-20 undergoes endoproteolytic cleavage into two disulfide-linked fragments whereas the released sPH-20 is not cleaved, suggesting the possible activity of a membrane-bound endoprotease on PH-20. We searched for a cDNA encoding sPH-20 but none was found. This result suggests that sPH-20 may arise from the enzymatic release of PH-20 from its membrane anchor, possibly at the time of acrosome reaction. PMID- 8724364 TI - Rat luteinizing hormone receptor messenger ribonucleic acid expression and luteolysis: inhibition by prostaglandin F2 alpha. AB - To elucidate the mechanisms by which prostaglandin F2 alpha (PGF2 alpha) permanently inhibits LH-dependent steroidogenesis during luteolysis, we investigated the effect on luteal LH receptor mRNA levels of the stable PGF2 alpha analogue cloprostenol injected into adult pseudopregnant rats on different days during the luteal period. After treatment, LH receptor mRNA expression was determined by RNase protection assay. Twelve hours after cloprostenol injection on Day 8 of pseudopregnancy, the luteal LH receptor mRNA levels were drastically reduced (0.95 +/- 0.18 fmol mRNA/microgram DNA, p < 0.01) as compared with those in untreated controls (12.3 +/- 1.3 fmol mRNA/microgram DNA) or in corresponding controls given an injection of saline (8.8 +/- 0.7 fmol mRNA/microgram DNA) (n = 6-8 per group). At 24 h the levels rose to 4.3 +/- 0.8 fmol mRNA/ microgram DNA but were still significantly decreased compared to control values. Forty-eight hours after cloprostenol injection, the luteal LH receptor mRNA levels were not significantly different from control levels; but if the rats received an injection every twelfth hour, levels were significantly decreased compared to those in controls. When PGF2 alpha was injected, LH receptor mRNA levels were reduced in the same manner as seen after cloprostenol injection. LH receptor mRNA of young corpora lutea (CL) (Day 3) was more resistant to down-regulation by cloprostenol than that of CL of the mid (Day 8)-or late (Day 11) luteal phase. On the eighth day of pseudopregnancy, serum progesterone levels were decreased at 0.5 h after cloprostenol injection and fell further at 3 h; serum 20 alpha dihydroprogesterone levels were first increased at 7 h after cloprostenol injection. We conclude that luteal LH receptor mRNA expression is under direct regulatory control by PGF2 alpha in a both time-and dose-dependent manner and thereby may decisively contribute to the inhibition of LH responsiveness during luteolysis. PMID- 8724365 TI - Proliferative activity in the different ovarian compartments in cycling rats estimated by the 5-bromodeoxyuridine technique. AB - The ovary is one of the most active proliferating tissues, and several methods for assessing cellular proliferation have been used to study follicular kinetics during the estrous cycle in the rat. In this study, we have estimated the proliferative activity of the different ovarian tissue compartments by using the 5-bromodeoxyuridine (BrdU) technique. Adult cyclic rats were studied 1 h after pulse-labeling with BrdU. Follicles were divided by size as small (mean diameter < 275 micrograms) or large (mean diameter > or = 275 micrograms). Small follicles were recorded into five classes: 1) primordial follicles (formed by flattened pre granulosa cells), 2) unilaminar primary follicles (which ranged from having at least one enlarged pre-granulosa cell up to having a complete layer of cuboidal granulosa cells), 3) multilaminar class a (having 1-2 layers of granulosa cells and measuring up to 75 micrograms in diameter), 4) multilaminar class b (76-150 micrograms), and 5) multilaminar class c (151-274 micrograms). Large follicles were recorded into classes 1-5 according to previous classifications [Osman P. J Reprod Fertil 1985; 73:261-270]. Primordial follicles and about 70% of unilaminar primary follicles were unlabeled, whereas all of the remaining classes of small and large follicles were labeled. The labeling index (percentage of BrdU-positive cells) increased in parallel with follicle size but showed a progressive decrease in large preovulatory follicles and a centripetal pattern of cell proliferation. Atretic follicles showed a significantly lower number of labeled cells than healthy follicles of the same size. The number of proliferating cells was not related to follicle size but was inversely correlated with the degree of atresia. Cyclic CL showed intense proliferative activity that was well correlated with functional activity. The number of labeled cells was maximal in metestrus, decreased in the succeeding days, and was very low in proestrus. Proliferating cells seemed to correspond to vascular cells lining blood vessels. These results indicate that the BrdU technique is a useful, accurate, and sensitive method for assessing cellular proliferation in the rat ovary. PMID- 8724366 TI - Identification and distribution of prostaglandin E receptors on porcine luteal cells. AB - This study examined whether pig luteal cells contain specific prostaglandin (PG) E receptors. Corpora lutea, from PG 600R -induced cycling prepubertal gilts or naturally cycling gilts, were dissociated enzymatically, and large, small, and mixed cells were obtained by elutriation. Preliminary experiments demonstrated that optimal [3H]PGE2 binding to mixed cells was obtained after incubation at 22 degrees C for 2 h in binding buffer containing 1% BSA at pH 7.38. In addition, [3H]PGE2 binding was displaced by prostaglandins in the order of PGE2 approximately or = to PGE1 > PGF2a > > PGI2. The mixed, large, and small cells were assayed for specific PGE2 binding by displacement analysis with [3H]PGE2 and varying concentrations of unlabeled PGE2. Each gilt was assigned to group A (n = 6) or group B (n = 3) on the basis of whether the serum progesterone concentrations in that pig had increased or decreased, respectively, from 2 days before ovary collection to the time of ovary collection. Luteal cells from PG 600R -induced cycling prepubertal gilts had similar (p > 0.1) affinities and numbers of PGE receptors. In group A, the mixed cells contained a PGE binding site with a Kd of 21.1 +/- 1.1 nM and a concentration of 5.0 x 10(5) sites per large cell. Affinities of PGE receptors were similar (p > 0.1) regardless of cell types and functional status. PGE receptor concentrations were higher on large cells than on small cells (p > 0.05) and in group A than in group B (p = 0.006). These data demonstrated that PGE receptors primarily exist on porcine large luteal cells and that the concentration of PGE receptors appeared to be related to corpus luteum functional status. PMID- 8724367 TI - Validation of radioimmunoassays for two salmon gonadotropins (GTH I and GTH II) and their plasma concentrations throughout the reproductivecycle in male and female rainbow trout (Oncorhynchus mykiss). AB - RIAs were developed for the two salmon gonadotropins (GTH I and GTH II) and used to measure plasma GTH throughout the life of the rainbow trout, Oncorhynchus mykiss. The RIA for GTH II was specific and sensitive (< 0.001% cross-reaction with GTH I, mean sensitivity = 0.26 +/- 0.02 ng/ml). The RIA for GTH I was less specific and less sensitive than the GTH II RIA (9.7% cross-reaction with GTH II, mean sensitivity = 2.34 +/- 0.23 ng/ml). In both males and females, the levels of GTH II remained undetectable (< 0.3 ng/ml) throughout most of the reproductive cycle, until shortly preceding spermiation/ovulation, when they began to rise. Concentrations of plasma GTH II were maximal at spermiation/ovulation. In both sexes, plasma profiles of GTH I differed from those of GTH II. The plasma GTH I concentration in females was elevated during early vitellogenesis. It then fell to a basal level shortly before ovulation and finally was elevated again at ovulation. In males, increases in plasma GTH I were seen a year before spermiation and again later, during the final stages of testicular growth. These results support the contention that GTH I mediates gonadal growth, whereas GTH II regulates the final stages of maturation and ovulation/spermiation. In rainbow trout, plasma profiles of GTH I and GTH II mimic the cycles of plasma FSH and LH, respectively, in the ovulatory cycle of higher vertebrates. PMID- 8724368 TI - Pervanadate mediated an increased generation of inositol phosphates and tension in rat myometrium. Activation and phosphorylation of phospholipase C-gamma 1. AB - Stimulation of [3H]inositol-labeled rat myometrial strips with pervanadate, formed by mixing orthovanadate and H2O2, induced a dose-dependent accumulation of [3H]inositol phosphates. Orthovanadate or H2O2 added alone had no effect. Pretreatment of myometrium with two tyrosine kinase inhibitors, namely genistein and tyrphostin 47 (at 100 microM), reduced pervanadate-stimulated inositol phosphate formation by 50%. Pervanadate induced a time-sequential formation of inositol phosphates in the order inositol trisphosphate, inositol bisphosphate, and inositol monophosphate. The inhibitory effect of genistein was observed at the level of the three inositol phosphates. Pervanadate induced contraction of the myometrium; the response was dose-dependent. H2O2 or orthovanadate was without effect. Pervanadate-mediated contraction was inhibited (50%) by genistein and tyrphostin 47 (100 microM). Western blot analysis, using anti-phosphotyrosine antibodies, revealed that phosphorylated proteins were present in detergent extracts from pervanadate-stimulated myometrium. Tyrosine phosphorylation was reduced by a preincubation with 100 microM genistein or tyrphostin 47. Phospholipase C-gamma1 was immunodetected in myometrial extracts and was identified as one of the substrates subject to tyrosine phosphorylation following pervanadate treatment. The results demonstrate that, in myometrium, protein tyrosine kinase/phosphatase activities controlled both phosphorylation and activation of phospholipase C-gamma1, contributing to the modulation of the generation of inositol phosphates and tension. PMID- 8724369 TI - Lipopolysaccharide inhibits in vitro luteinizing hormone-stimulated rat ovarian granulosa cell estradiol but not progesterone secretion. AB - Endotoxin, known as lipopolysaccharide (LPS), is a component of gram-negative bacterial cell walls and is a potent immunostimulator, inducing the release of several cytokines, such as tumor necrosis factor alpha (TNF alpha) and interleukins (IL) 1, 6, and 8. A previous study with immature rats revealed that exogenous administration of LPS inhibits ovarian estradiol secretion in response to eCG. The present study was undertaken in order to determine whether LPS could directly inhibit rat granulosa cell (GC) steroid secretion. GC were collected and purified from 26-day-old hypophysectomized female rats (hypophysectomy on Day 23). Purified GC were highly responsive to FSH (1-100 ng/ml), leading to increased estradiol, progesterone, and cAMP accumulation in culture media. GC were also capable of binding 125I-labeled hCG and were responsive to LH stimulation. Treatment of GC with LPS (1-100 ng/ml) led to a significant (p < 0.01) dose-dependent decrease in LH-stimulated estradiol accumulation in culture media (maximum 75% inhibition). However, treatment of GC with LPS had no significant effect on FSH-stimulated progesterone or estradiol, or LH-stimulated progesterone accumulation in culture media. GC stimulated with 8-bromo cAMP were also insensitive to the effects of LPS. LPS had no significant effect on 125I labeled hCG binding to GC homogenates, nor did it have any significant effect on FSH or LH-stimulated cAMP accumulation. Treatment of both FSH and LH-stimulated GC with LPS was associated with an increase in IL-6 bioactivity in culture media. This effect could be blocked with the nonreceptor tyrosine kinase inhibitor herbimycin A. TNF alpha bioactivity was undetectable with or without LPS challenge. Direct challenge of GC with recombinant murine IL-6 had no effect on either FSH or LH-stimulated estradiol whereas TNF alpha inhibited FSH-stimulated estradiol secretion. Collectively, these results suggest that the inhibitory effects of LPS were not mediated by either IL-6 or TNF alpha. Treatment of GC with the epidermal growth factor receptor tyrosine kinase inhibitor, tyrphostin A46, blocked the inhibitory effects of LPS on steroid secretion and was associated with an increased cAMP accumulation in culture media. The results indicate that LPS inhibits in vitro GC estradiol secretion. This effect appears to be restricted to the LH-stimulated aromatization of androgens to estrogen and may involve a tyrosine kinase signaling pathway. PMID- 8724370 TI - Microtubule organization in porcine oocytes during fertilization and parthenogenesis. AB - Microtubule configurations in porcine oocytes after sperm penetration or after artificial activation by electrical stimulation were imaged by immunocytochemistry and laser scanning confocal microscopy. Soon after sperm penetration, an aster was seen adjacent to the incorporated sperm head. Polyspermic penetrations led to the presence of multiple sperm asters in association with each sperm. The sperm aster enlarged and, at the time of pronuclear apposition, filled the cytoplasm. After male and female gamete union, the microtubule matrix was reduced. At the mitotic metaphase stage, microtubules were detected in the spindle, which was anastral and fusiform. At anaphase, asters assembled at each spindle pole, and at telophase, large asters filled the cytoplasm. Artificial activation by electrical stimulation induced in the cytoplasm a dense network of microtubules, which seem to be involved in proper positioning of the female pronucleus. At mitotic metaphase, microtubules were concentrated around the chromatin. The results of experiments using taxol, a microtubule stabilizing agent, suggest that maternal centrosomal material is present in the mature porcine oocyte as dispersed undetectable material that can form a microtubule network after parthenogenetic activation. However, at fertilization, the paternal centrosome collects centrosomal material to form a sperm aster. These results suggest that the functional centrosome that forms during fertilization is a result of the blending of paternal and maternal centrosomal components. PMID- 8724371 TI - Stimulation of human sperm during capacitation in vitro by an adenosine agonist with specificity for A2 receptors. AB - The effects of an adenosine agonist with specificity for A2 receptors, on human sperm prepared for in vitro fertilization (IVF), were examined to verify physiological effects and possible pharmacological use. 5' -N-ethyl carboxamidoadenosine (NECA), when added at 100 microM over 30 min in B2 medium, did not induce a spontaneous acrosome reaction after 0, 3, and 6 h capacitation in B2, nor did it modify sperm motility. However, NECA increased the number of capacitated spermatozoa able to respond (p < 0.05) to A23187 (10 microM) after 6 h preincubation in B2 medium. This effect was associated with an increase in cAMP production, which was measured by RIA after 10 and 20 min incubation with NECA in uncapacitated sperm, and with changes in the kinetics of protein tyrosine phosphorylation as revealed by Western blot. Phosphorylation of a 95-kDa protein was enhanced by NECA in uncapacitated sperm and inhibited in capacitating sperm (incubated 1 h in B2), whereas phosphorylation of a 50-kDa protein was systematically enhanced whatever the preincubation time. NECA can stimulate uncapacitated human sperm via cAMP production and protein phosphorylation/dephosphorylation without inducing an acrosome reaction or influencing motility. Cyclic AMP-dependent protein kinase A seems to positively control protein phosphorylation involved in human capacitation. Adenosine present in the tubal fluid or produced by the spermatozoon itself may influence capacitation in vivo through sperm A2 receptors. In cases of male infertility, use of NECA in sperm handling for IVF should be evaluated as a means to improve capacitation without increasing the possibility of a premature spontaneous acrosome reaction. PMID- 8724372 TI - Presence of organic osmolytes in maturation medium enhances cytoplasmic maturation of porcine oocytes. AB - The effects of organic osmolytes on cytoplasmic maturation of porcine oocytes were examined in maturation medium (modified Whitten's medium) containing various NaCl concentrations. The presence of organic osmolytes, such as taurine and sorbitol, at 6 and 12 mM in maturation medium containing 68.49 or 92.40 mM NaCl increased oocyte glutathione content. Microfilament organization in oocytes was disrupted in maturation medium containing the higher level of NaCl (92.40 mM). However, supplementation with 12 mM sorbitol to the medium reduced the severity of the abnormality. Early embryonic development in vitro to the blastocyst stage was 8.3 +/- 0.9% for oocytes matured in modified Whitten's medium (68.49 mM NaCl) supplemented with 12 mM sorbitol, and 7.9 +/- 0.8% in modified NCSU23 medium (containing 108.73 mM NaCl, 7 mM taurine, 5 mM hypotaurine, and 1 mM glutamine), compared to 4.7 +/- 0.6% in modified Whitten's medium (68.49 mM Na Cl), which did not contain organic osmolytes. These results indicate that the presence of organic osmolytes, such as sorbitol and taurine, reduces the detrimental effects of high NaCl concentration in media used for the maturation of porcine oocytes. This effect is reflected by oocyte glutathione content and microfilament organization at the end of maturation and early development following in vitro maturation and in vitro fertilization. PMID- 8724373 TI - Role of epidermal growth factor in bovine oocyte maturation and preimplantation embryo development in vitro. AB - Epidermal growth factor (EGF) has been shown to have a positive effect during in vitro maturation (IVM) and has been reported in follicular fluid at levels capable of stimulating meiosis in a variety of species. The aim of the present work was to study the effect on subsequent development of EGF present in defined medium during bovine 1) oocyte maturation or 2) embryo culture. The presence of EGF during IVM, irrespective of concentration (1, 10, 100 ng/mg), stimulated cumulus expansion and significantly increased the proportion of oocytes attaining metaphase II, the rate of cleavage, and the proportion of embryos reaching the 5- to 8-cell stage at 72 h postinsemination. Blastocyst rates on Days 7 and 9 were also significantly improved for oocytes matured in the presence of EGF (10% vs. 18-24% on Day 7 and 21% vs. 31-32% on Day 9, for Tissue Culture Medium 199 [M199] and M199 + EGF, respectively). The presence of fetal calf serum (FCS) during IVM resulted in similarly elevated rates of development. There was no cumulative effect when EGF and FCS were present together during IVM. The presence of EGF also altered the pattern of proteins neosynthesized during maturation. The maturation-promoting effect of EGF was evident for denuded oocytes also, suggesting that EGF may act, at least in part, directly on the oocyte. Immunofluorescence studies revealed the EGF receptor on immature cumulus-oocyte complexes. When present during postfertilization culture in defined medium (synthetic oviduct fluid), EGF stimulated development in comparison to that of the control but could not replace serum. The results suggest a physiological role for EGF in the regulation of bovine oocyte maturation and development. PMID- 8724374 TI - Cell-cell interactions related to reproductive biology: report of National Institutes of Health Workshop, June 11, 1995, Bethesda, Maryland. PMID- 8724375 TI - Anorexia during acute and chronic disease. AB - Anorexia is associated with disorders of all systems. Anorexia represents a consistent clinical manifestation during acute and chronic pathophysiological processes (infection, inflammation, injury, toxins, immunological reactions, malignancy and necrosis). Anorexia during disease can be beneficial or deleterious depending on the timing and duration. Temporary anorexia during acute disease may be beneficial to an organism since a restriction in the intake of micro- and macro-nutrients will inhibit bacterial growth. Long-term anorexia during chronic disease, however, is deleterious to an organism and may be associated with cachexia, which can ultimately result in death. Various mechanisms participate in the anorexia observed during disease, including cytokine action. Anorexia induced by cytokines is proposed to involve modulation of hypothalamic-feeding associated sites, prostaglandin-dependent mechanisms, modifications of neurotransmitter systems, gastrointestinal, metabolic, and endocrine factors. In addition, the anorexia-cachexia syndrome is multifactorial and may involve chronic pain, depression or anxiety, hypogeusia and hyposmia, chronic nausea, early satiety, malfunction of the gastrointestinal system, metabolic alterations, cytokine action, production of other anorexigenic substances and/or iatrogenic causes (chemotherapy, radiotherapy). Cachexia may result not only from anorexia and a decreased caloric intake, but also from malabsorption and losses from the body (ulcers, hemorrhage, effusions), or a change in body metabolism. Research has focused on potential interventions to modify anorexia during disease and the anorexia-cachexia syndrome. Nutritional modifications and the use of specific steroids (such as megestrol acetate) are being tested in the clinical setting. Understanding the specific mechanisms responsible for anorexia during disease as well as their interactions is essential to develop interventions for the control of anorexia (during a critical time in a specific disease), and to devise less toxic immunotherapeutic regimens using cytokines. PMID- 8724376 TI - Elimination rate of fat emulsion particles from plasma in Japanese subjects as determined by a triglyceride clamp technique. AB - The elimination rate of emulsion triglyceride (TG) from plasma was investigated in Japanese subjects by a plasma TG clamp technique. Two different studies were performed. In Study 1, a lipid emulsion (20% long-chain triglyceride emulsion: LCT) was infused into a healthy research associate to achieve a certain concentration of TG in the plasma. Thereafter, the infusion rate was adjusted to maintain the chosen concentrations of TG in plasma (namely, 4-5 mmol/L, 3-4 mmol/L, and approximately 2 mmol/L) over a period of 160 min by measuring plasma TG concentrations at 10-min intervals. Concentrations of TG in plasma were clamped within 2.13 +/- 0.13 mmol/L by an infusion rate of 0.10 g.kg-1.h-1, within 3.34 +/- 0.20 mmol/L by an infusion rate of 0.14 g.kg-1.h-1, and within 4.46 +/- 0.22 mmol/L by an infusion rate of 0.11 g.kg-1.h-1. The mean rate of infusion of emulsified TG that had maintained the steady concentrations of TG in plasma was limited to the very narrow range of 0.12 +/- 0.02 g of TG.kg-1.h-1 regardless of the chosen concentration of TG in plasma. Concentrations of nonesterified fatty acids (NEFA) also remained at a fixed level of 1.378 +/- 0.103 mEq/L regardless of the chosen concentration of TG in the plasma. Study 2 was undertaken to determine whether plasma TG concentration reached a plateau during a period when emulsion TG was infused into three different subjects at a rate of 0.12 g.kg-1.h-1. The plasma TG concentrations were steady at a level of 2.04 +/- 0.32 mmol/L, and the plasma NEFA concentrations remained at a fixed level of 1.33 +/- 0.13 mEq/L, over a period of 160 min after 50-min priming infusion. These results indicate that the plasma TG elimination rate was limited to the narrow range of 0.12 +/- 0.02 g.kg-1.h-1 when the fat emulsion was infused into Japanese subjects in a steady state. However, the plasma TG elimination rate in Japanese subjects appeared to be lower than that of Europeans. This may be due to a difference in lipoprotein lipase activity caused by different dietary habits, namely, a lower fat intake. PMID- 8724377 TI - Low ceruloplasmin levels during recovery from major burn injury: influence of open wound size and copper supplementation. AB - Copper (Cu) status is often judged by the plasma level of its chief transport protein, ceruloplasmin (Cp). Only copper deficiency and heredity are known to decrease circulating Cp. Cp is an acute-phase responsive protein in trauma and it is also induced by Cu supplementation. Despite this, plasma concentrations of Cp remain low during the acute recovery from major burn injury. The high provision of vitamin C typically used in burn patients may influence these observations when an indirect oxidase activity assay is used. We employed a radial immunodiffusion (RID) assay specific for the Cp protein as well as an indirect oxidase assay for Cp in a series of 11 burned children who were supplemented with both Cu and vitamin C, either enterally or parenterally. Our findings confirm that low Cp is a characteristic of the acute recovery from major burns. The oxidase assay is shown to be valid for very low Cp levels even during high vitamin C provision. When these data are combined with our previously reported series, a strong relationship between the size of the open wound area and the amount of circulating Cp is demonstrated. Copper supplementation by either the enteral or parenteral routes is only marginally successful in restoring Cp toward normal levels. PMID- 8724378 TI - NO2/NO3 and cytokine plasma profiles under different postoperative parenteral nutrition regimens. AB - Both nitric oxide and cytokines are considered mediators of the acute-phase response in humans, and their early postoperative period plasma levels have been found to be of prognostic value. On the other hand, it has been suggested that the fatty emulsions used in total parenteral nutrition (TPN) may induce changes in macrophage function. In the present study we investigated the postoperative evolution of tumor necrosis factor-alpha (TNF-alpha), interleukin-1 (IL-1), interleukin-6 (IL-6), and nitrate/nitrite plasma levels under three different TPN regimens. Twenty-one patients diagnosed with upper digestive tract neoplasm, without preoperative TPN, and having undergone radical surgery, were randomly assigned to three groups: Group I, all nonprotein calories supplied by hypertonic glucose solution: Group II, 55% of the nonprotein calories supplied by glucose and 45% by 20% long-chain triacylglycerides emulsion (LCT) (Intralipid 20%, Kabi Pharmacia); Group III, same as Group II, but a 20% emulsion of a mixture of medium-chain and long-chain triacylglycerides (MCT/LCT) (Lipofundina MCT/LCT 20%, B. Braun) was used instead of LCT. Blood samples were obtained on postoperative Days 1-5 and 10, 3 h after ending the lipid infusion. In all the three groups IL 1, IL-6, and TNF-alpha levels rose after surgery, peaking at Day 2, whereas NO2/NO3 levels had their peak at Day 3. Day-to-day comparison of plasma levels of cytokines and NO2/NO3 between the investigated groups did not show any statistical significance. Differences between group means were not found when the areas under the curve over the first 5 postoperative days were compared (1.72 +/- 0.25, Group I; 1.88 +/- 0.34, Group II; and 2.52 +/- 0.50, Group III, for TNF alpha; 1.79 +/- 0.12, Group I; 1.92 +/- 0.18, Group II; and 1.50 +/- 0.12, Group III, for NO2/NO3). We conclude that the different parenteral nutrition regimens studied do not evoke alterations in cytokine and NO2 + NO3 levels in the patient groups investigated in this study. PMID- 8724379 TI - Influence of medium-chain triglyceride-based lipid emulsion on rat polymorphonuclear cell functions. AB - Intravenous lipid emulsions are used as energy and essential fatty acids sources. There are controversial reports postulating in vitro and in vivo inhibitory effects of long-chain triglycerides (LCT) upon the blood polymorphonuclear leukocytes (PMNL) functions. In the present study the in vivo and in vitro effects of LCT and a physical mixture of medium- and long-chain triglycerides (MCT/LCT) emulsions were investigated on select PMNL functions, i.e., chemotaxis, phagocytosis, and bacterial killing. Blood from 20 rats was incubated with LCT, MCT, MCT/LCT, and saline, respectively. MCT-containing emulsions exhibited an inhibitory effect on all PMNL functions investigated, whereas LCT exerted an effect on the phagocytic index only. The administration of a parenteral supply of LCT, MCT/LCT, and saline for 30 h followed by saline infusion for 14 h in discontinuous mode did not influence any of the investigated PMNL functions. Similarly, continuous infusion over 44 h at increasing infusion rates up to 1.5 mL/h did not affect the PMNL functions. The obvious difference between in vitro and in vivo response of the PMNL model emphasizes the necessity for continuous monitoring of in vivo conditions. Appropriate interpretation of the data requires continuous circumspection and consideration of trials in a clinical setting. PMID- 8724381 TI - Nutritional and surgical management of perforation of the esophagogastric junction. PMID- 8724380 TI - Flavonoids extracted from fonio millet (Digitaria exilis) reveal potent antithyroid properties. AB - Digitaria exilis (fonio) is a tiny variety of millet commonly eaten by inhabitants of semiarid regions. A sample of fonio collected right in the middle of a severely iodine-depleted goitrous endemic was submitted to phytochemical investigations in order to assess the potential contributory roles played by vegetable molecules to the goitrogenic processes. The total content of flavonoids amounts to 500 mg/kg of the edible whole cereal grains. Their extraction and identification fail to detect the C-glycosylflavones described in other millet varieties but point out the presence of apigenin (A = 150 mg/kg) and of luteolin (L1 = 350 mg/kg). Ten percent of A and 80% of L1 are present in free form, whereas the remaining 90% of A and 20% of L1 are bound as O-glycosylflavones. Both A and L1 aglycones manifest strong anti-thyroid peroxidase (TPO) activities, resulting in a significant reduction of the hormonogenic capacity of this enzyme. In addition, L1 significantly depresses the cyclic AMP phosphodiesterase, implying a concomitant overproduction of the thyrotropin-dependent nucleotide. These last unreported data are regarded as counteracting to some extent the TPO mediated goitrogenic properties of L1. Since fonio is devoid of other molecules likely to interfere with the thyroid function, our results are directly and casually attributed to A and L1 found in the customary diet. PMID- 8724383 TI - Changing from fasting motor activity to the fed pattern in the gut: how it happens. PMID- 8724382 TI - From laboratory to bedside: new strategies in the treatment of malnutrition in cancer patients. AB - The objective of this review is to present and discuss the current and the possible future perspectives of artificial nutrition in preventing and treating cancer cachexia. English-language studies published in the last 25 years were retrieved using MEDLINE, bibliographies and consultation with experts. MEDLINE search terms included "cancer," "malnutrition," and "nutritional support." In vivo and in vitro controlled studies were selected, and data from each study were independently abstracted by the authors. Data obtained indicate that cancer cachexia is a common manifestation of advanced malignant disease, characterized by marasmus due to impaired nutrient metabolism, weight loss, and anemia. In patients, the presence of cachexia increases morbidity and mortality, delays or impedes the delivery of the adequate treatment, reduces the efficacy of anti neoplastic therapy, and impairs patient's quality of life. Indirect evidence indicates that cancer cachexia is mediated by circulating cytokines, produced by the host in response to the presence of the malignancy. In the past two decades, nutritional support was used to counteract the detrimental effect of cancer cachexia on the host, but with limited success. More recently, the better understanding of the mechanisms of cancer cachexia and the identification of pharmacologic effects provided by specific nutrients, has formed the rationale for a "new" mode of nutrition therapy/nutritional support, adding pharmacotherapy to adjunctive initial care. PMID- 8724384 TI - Nutritional alterations in high-risk groups in relationship to HIV-1 disease progression. PMID- 8724385 TI - Nutritional support of head-injured patients. PMID- 8724386 TI - On the biologic significance of vitamin C interaction with ceruloplasmin. PMID- 8724387 TI - Flavonoids and the thyroid. PMID- 8724388 TI - Interaction between nutrition, interleukins, and the NO-arginine system. PMID- 8724390 TI - Logistic regression. PMID- 8724389 TI - Recent outcomes research in Britain. PMID- 8724391 TI - The silence of the lions. PMID- 8724392 TI - Roles and functions of the hospital pharmacist on the nutrition support team. PMID- 8724393 TI - Roles and functions of the hospital pharmacist on the nutrition support team. PMID- 8724394 TI - Defining dietetics practice for today and tomorrow: an international perspective. PMID- 8724395 TI - Device selection: a critical strategy in the reduction of catheter-related complications. PMID- 8724396 TI - Stool cultures for Shigella spp: improved specificity by using MacConkey agar with xylose. AB - A total of 678 stool specimens were cultured on four different agars: on xylose lysine-desoxycholate agar (XLD), MacConkey agar (Mac), MacConkey agar supplemented with xylose (Mac-X), and Hektoen enteric agar (HE). Isolation rates for shigellae were 77% on HE, 86% on Mac and Mac-X, and 91% on XLD. The specificities of the media were 61% for Mac, 75% for HE, and 78% for XLD and Mac X. After overnight incubation, Mac-X is much easier to read than XLD, which requires incubation for at least 22 hours. Based on these results and also on the practical aspect that incubation for 22-21 hours does not fit well in our schedule, we now use Mac-X whenever shigellae need to be looked for (i.e. mainly patients with recent travel to tropical countries). As compared to our previous procedure the workload in the laboratory could be reduced by about 20%. PMID- 8724397 TI - Rapid detection of group B streptococcal antigen from vaginal specimens using a new Optical ImmunoAssay technique. AB - A total of 531 vaginal specimens were used to evaluate a new Optical ImmunoAssay (OIA) screening technique for the rapid detection of group B streptococcal antigen. The results of the OIA test, the ICON Strep B membrane immunoassay (Hybritech ICON), and conventional culture on sheep blood agar (direct TSA) were compared to broth enhanced culture. Results obtained from the OIA test, ICON, and direct TSA yielded 72, 39, and 68 positives, respectively, as compared to 100 positives using the Lim broth culture method as the standard. The Optical ImmunoAssay technique is as sensitive as the conventional plating method and is capable of providing results in 30 minutes. PMID- 8724398 TI - Legionella urinary antigen testing: potential impact on diagnosis and antibiotic therapy. AB - Although Legionella is an important cause of severe pneumonia, difficulty still exists in its diagnosis. Because at least 80% of patients with legionellosis excrete the Legionella antigen in their urine, various methods have been investigated for urinary antigen detection. Specificity for these methods has been reported to be 100%, and sensitivity has been shown to vary between 70 and 100%. The advantages of these methods include ease of specimen collection, the ability to obtain large quantities of specimen for concentration, the ability to detect antigen after initiation of antibiotic therapy, and the ability to obtain results quickly. Disadvantages include the ability to only reliably detect urinary L. penumophila serogroup 1 antigen and the inability to diagnose relapse or reinfection due to persistence of antigen excretion. Of the commercially available detection methods, the polyclonal enzyme-linked immunosorbent assay (ELISA) appears to be most efficient. Its use with routine Legionella screening procedures should be considered in target populations, with the intent of early diagnosis and antibiotic therapy streamlining. PMID- 8724399 TI - Lack of requirement for prolonged incubation of Septi-Chek blood culture bottles in patients with bacteremia due to fastidious bacteria. AB - A questionnaire survey for which there were 538 respondents from American clinical microbiology laboratories was used to assess the need for prolonged incubation of Septi-Chek blood culture bottles for the recovery of HACEK bacilli, Brucella species, Francisella tularensis, and nutritionally deficient streptococci from blood. Among a total of 219 reported isolates of these bacteria, in only 6 cases (2.7%) was incubation longer than 7 days required. Only 2 of 136 patients (1.5%) were noted to be bacteremic exclusively following incubation of Septi-Chek blood culture bottles for periods of greater than 7 days. It appeared from the results of this study, that incubation of Septi-Chek blood culture bottles for 7 days is usually sufficient in patients suspected of having bacteremia due to fastidious bacteria. PMID- 8724400 TI - Comparison of five different susceptibility test methods for detecting antimicrobial agent resistance among Haemophilus influenzae isolates. AB - The detection of antimicrobial agent resistance among ninety-eight Haemophilus influenzae isolates was assessed by six different antibiotic test methods: agar dilution on Mueller-Hinton agar supplemented with 5% lysed horse blood (MH-LHB), E-test using both Haemophilus test medium (HTM) agar and chocolate Mueller-Hinton (CMH) agar plates, Vitek Haemophilus susceptibility cards, and three overnight microdilution systems that included two commercial systems, Micro-Media and MicroScan, and the reference broth microdilution method using HTM broth. Agents tested in the study included ampicillin, amoxicillin/clavulanic acid (A/C), cefaclor, cefuroxime, cefotaxime, ceftriaxone, chloramphenicol, and trimethoprim/sulfamethoxazole. Both the reference HTM microbroth dilution method and agar dilution correctly classified all nine of the beta-lactamase negative ampicillin resistant (BLNAR) isolates. Each of the other test methods failed to detect one of the BLNAR strains, either because of growth failure (Micro-Media and MicroScan) or miscategorization of an isolate as susceptible (E-Test HTM, E Test CMH, and Vitek). None of the test methods detected all six isolates identified as A/C resistant by HTM microbroth dilution. Of the remaining antimicrobials tested, ampicillin and cefuroxime yielded data that could be compared by all test methods. The very major, major, and minor errors for these two antimicrobials in comparison to the reference HTM microdilution method were as follows: Micro-Media (1.7%, 0%, and 4.8%); MicroScan (11.9%, 0%, and 8.1%); E Test HTM (1.6%, 0%, and 2.0%); E-Test CMH (1.6%, 1.6%, and 4.6%); Vitek (8.1%, 0%, and 3.1%); and agar dilution on MH-LHB (0%, 0%, and 4.6%). Micro-Media and MicroScan panels failed to support the growth of 4.1% and 5.1% of the isolates, respectively. PMID- 8724401 TI - Susceptibility testing interpretive criteria for levofloxacin when testing respiratory pathogens, Haemophilus influenzae and Moraxella catarrhalis. AB - The more active L-isomer, levofloxacin, of the racemic ofloxacin mixture has been under development for therapeutic use. In this study, we evaluated the activity of ofloxacin, levofloxacin, and D-ofloxacin against the fastidious respiratory tract pathogens Haemophilus influenzae and Moraxella catarrhalis. Levofloxacin was two-fold more active than ofloxacin against H. influenzae (MIC90, 0.015 microgram/ml), and D-ofloxacin was least active (MIC90, 1 microgram/ml). For M. catarrhalis the MIC90 values were 0.03 microgram/ml, 0.06 microgram/ml, and 2 micrograms/ml for levofloxacin, ofloxacin, and D-ofloxacin, respectively. For disk diffusion susceptibility testing, Chocolate Mueller-Hinton agar (CMH) was considered preferable to Haemophilus test medium (HTM) because it supported the growth of all of 105 H. influenzae strains whereas five strains failed to grow on HTM. In addition, the margins of the zones of inhibition were more distinct on CMH and the Haemophilus species strains with elevated fluoroquinolone MICs were readily distinguished. The superior growth on CMH was reflected in a reduction of inhibition zone diameters of 2-3 mm relative to the inhibition zone diameters on HTM. The previously proposed interpretive criteria for the 5 microgram disk diffusion susceptibility test (susceptible at > or = 17 mm) results in complete categorical agreement with the reference microdilution broth method for M. catarrhalis on Mueller Hinton agar and for H. influenzae on HTM and CMH. However, the minimum diameter of the zone of inhibition recorded for a member of the dominant population of either species was considerably greater (25 mm) than 17 mm on any of the media tested. PMID- 8724402 TI - Candida albicans osteomyelitis of the zygomatic bone. A distinctive case with a possible peculiar mechanism of infection and therapeutic failure with fluconazole. AB - This report describes a distinctive case of zygomatic candidiasic osteomyelitis in a diabetic patient with oral candidiasis and malar ulceration secondary to topic 5-fluoroacil toxicity that eventually exposed part of the underlying bone. The mechanism of infection may have been self-inoculation of spores from muguet plaques on the oral mucosa to the exposed bone tissue by hand contact. Such a mechanism of bone infection probably should be considered in patients who frequently have oral candidiasis (diabetes, malignancies, and HIV infection) and open lesions of the skin and soft tissues. Treatment with fluconazole was ineffective, but amphotericin B was curative. PMID- 8724403 TI - Antibodies that react with the capsular polysaccharide of Vibrio vulnificus are detectable in infected patients, and in persons without known exposure to the organism. AB - In serious infections with Vibrio vulnificus, IgG antibodies to the capsular polysaccharide of the infecting strain were demonstrable in patient serum. It was not possible to show that persons with probable increased exposure to V. vulnificus (shellfish industry workers) had increased levels of antibodies to any one of three capsular types tested when compared with persons who would be expected to have had minimal exposure to the organism (Seventh Day Adventists). Antibodies that reacted with the capsular polysaccharides were demonstrable in persons without a history of V. vulnificus infection, suggesting that cross reacting antibodies are present in the general population. PMID- 8724404 TI - Multi-center validation of proposed disk diffusion susceptibility testing interpretive criteria for lomefloxacin using more than 1,500 clinical isolates. AB - The accuracy of disk diffusion susceptibility testing of lomefloxacin was evaluated using 1,555 recent clinical isolates from ten medical centers. The isolates were rapidly growing nonfastidious aerobic species (1,501 isolates) and Haemophilus species (54 isolates), each found as an indicated species in the product package insert. Applying the recently proposed modification of disk diffusion interpretive criteria (susceptible at > or = 20 mm and resistant at < or = 16 mm), absolute categorical agreement for nonfastidious aerobes (1,501 strains) was 95.5% with 0.5% very major and 0.1% major errors (error rates calculated using all tested strains as the denominator). The intermethod discord (MIC vs disk diffusion) was 0.5%. This contrasts to the current NCCLS recommended criteria (susceptible at > or = 22 mm and resistant at < or = 18 mm) where the absolute categorical agreement was significantly less (89.6%) with 0.2% very major, and 0.3% major errors, and the intermethod discord was 7.1%. For Haemophilus species (54 strains), intermethod agreement was complete using either the current NCCLS interpretive criteria or the modified criteria. These multicenter (ten laboratories) data support the acceptance of the proposed modification of disk diffusion interpretive criteria for 10-microgram lomefloxacin disks. PMID- 8724405 TI - Minimal residual epithelial cancer: diagnostic approaches and prognostic relevance. PMID- 8724407 TI - Quantification of white matter and gray matter volumes from T1 parametric images using fuzzy classifiers. AB - White matter (WM) and gray matter (GM) were accurately measured using a technique based on a single standardized fuzzy classifier (FC) for each tissue. Fuzzy classifier development was based on experts' visual assessments of WM and GM boundaries from a set of T1 parametric MR images. The fuzzy classifier method's accuracy was validated and optimized by a set of T1 phantom images that were based on hand-detailed human brain cryosection images. Nine sets of axial T1 images of varying thickness equally distributed throughout the brain were simulated. All T1 data sets were mapped to the standardized FCs and rapidly segmented into WM and GM voxel fraction images. Resulting volumes revealed that, in most cases, the difference between measured and actual volumes was less than 5%. This was consistent throughout most of the brain, and as expected, the accuracy improved to generally less than 2% for the 1-mm simulated brain slices. PMID- 8724406 TI - Magnetic resonance imaging of the brain with gadopentetate dimeglumine-DTPA: comparison of T1-weighted spin-echo and 3D gradient-echo sequences. AB - Short TR, short TE, high resolution, 3D gradient-recalled echo (GRE) imaging was evaluated for lesion detection in the brain. High resolution 3D GRE data acquisition was used to reduce partial volume effects and flow artifacts, to better visualize smaller structures, to minimize signal losses caused by field inhomogeneities, and to allow better image reformatting. Spin-echo (SE) and 3D GRE approaches were compared for lesion detection after the administration of an MR contrast agent, gadopentetate dimeglumine. Preliminary clinical studies demonstrated that the signal-to-noise ratio (SNR) in each slice of the GRE scan was worse than that of the SE scan because of the much thicker slices acquired with the SE technique. However, by averaging two adjacent 3D slices, the SNR of the two methods was essentially equivalent. In the averaged GRE slices, large lesions were seen just as well as in the SE images. More importantly, small lesions were better visualized in the thin 3D GRE images than in the thick SE images for the lesions studied in this work and the protocols used. These observations were confirmed by theoretical simulations. PMID- 8724408 TI - Classification of 1H MR spectra of human brain neoplasms: the influence of preprocessing and computerized consensus diagnosis on classification accuracy. AB - We study how classification accuracy can be improved when both different data preprocessing methods and computerized consensus diagnosis (CCD) are applied to 1H magnetic resonance (MR) spectra of astrocytomas, meningiomas, and epileptic brain tissue. The MR spectra (360 MHz, 37 degrees C) of tissue specimens (biopsies) from subjects with meningiomas (95; 26 cases), astrocytomas (74; 26 cases), and epilepsy (37; 8 cases) were preprocessed by several methods. Each data set was partitioned into training and validation sets. Robust classification was carried out via linear discriminant analysis (LDA), artificial neural nets (NN), and CCD, and the results were compared with histopathological diagnosis of the MR specimens. Normalization of the relevant spectral regions affects classification accuracy significantly. The spectra-based average three-class classification accuracies of LDA and NN increased from 81.7% (unnormalized data sets) to 89.9% (normalized). CCD increased the classification accuracy of the normalized sets to an average of 91.8%. CCD invariably decreases the fraction of unclassifiable spectra. The same trends prevail, with improved results, for case based classification. Preprocessing the 1H MR spectra is essential for accurate and reliable classification of astrocytomas, meningiomas, and nontumorous epileptic brain tissue. CCD improves classification accuracy, with an attendant decrease in the fraction of unclassifiable spectra or cases. PMID- 8724409 TI - A model for detecting early metabolic changes in neonatal asphyxia by 1H-MRS. AB - In newborn rabbits, the early cerebral metabolic changes caused by hypoxic ischemic (H-I) insult was examined by using volume localized 1H-MRS (STEAM). Partial ischemia was caused by unilateral carotid artery ligation, and hypoxia was induced by 10% oxygen inspiration for 150 minutes. Lactate immediately increased after hypoxia induction and almost disappeared 120 to 150 minutes after removal of hypoxia in both H-I and hypoxia-only experiments. Lactate production correlated well with decrease of the blood oxygen saturation. More lactate was produced on ischemic side 50 minutes post-hypoxia induction in H-I study. Ischemia alone did not cause any significant lactate production. Lactate caused by hypoxia can be dynamically monitored by localized 1H-MRS. Existence of regional ischemia can induce greater anaerobic glycolysis and may affect the pattern of brain injury under hypoxia. 1H-MRS is a sensitive tool to detect the acute metabolic change caused by H-I insult. PMID- 8724410 TI - Localized 2D J-resolved 1H MR spectroscopy of human brain tumors in vivo. AB - Application of two-dimensional (2D) J-resolved MR spectroscopy, fully localized in three dimensions to monitor the metabolites in human brain tumors in vivo on a whole body MR scanner is presented. A modified PRESS sequence with [90 degrees - 180 degrees - t1/2 - 180 degrees - t1/ 2-acquisition] was used for voxel localization (2D J point-resolved spectroscopy [PRESS]); chemical shift selective (CHESS) sequence was used for suppression of water. The incremental delay (t1/2) added to the intervals before and after the last slice-selective 180 degrees RF pulse allowed the monitoring of the J-evolution in a localized 2D NMR spectrum. The addition of the second frequency dimension in 2D J-resolved spectroscopy to encode the indirect spin-spin coupling allowed the visualization of lactate peaks not observed in the 1D MR spectrum because of severe overlap with lipid peaks. 2D spectra of a two-layer phantom with 100 mM alanine and corn oil and also from three patients with tumors are presented here. The 2D spectra show that the J coupled lactate peaks could be separated even when the lipids peaks severely overlap. PMID- 8724411 TI - MR perfusion imaging of the kidney pre- and post-dipyridamole stress. AB - Animal studies have demonstrated that renal MR contrast enhancement depends on the timing of image acquisition. Limited human studies have demonstrated effects of dipyridamole (DP) on total renal perfusion. This study assessed the effect of DP on total and regional renal perfusion using gated perfusion MRI for patients undergoing DP stress. Five subjects with no evidence of renal ischemia were examined at rest and after DP stress. Rest MRI images in the left kidney were acquired using electrocardiogram (ECG)-gated MR: turbo fast low-angle shot (FLASH); echo time (TE) = 12, repetition time (TR) = 6, flip angle = 12, inversion time (TI) = 100) 10 to 45 seconds after injection of gadopentetate dimeglumine. Stress was induced in the MRI scanner (DP, .56 mg/kg over 4 minutes) followed by stress MRI after a second bolus of gadopentetate dimeglumine in the same position and identical time intervals. MR signal in the whole left kidney and renal medulla and cortex pre- and post-DP demonstrated a 70% depression of total renal perfusion with relative preservation of cortical perfusion at the expense of medullary perfusion. Post-DP MR images demonstrated a decrease in cortical perfusion with an additional 29% depression of medullary perfusion (P < .001) with respect to cortical perfusion. Turbo FLASH MRI can provide adequate time and spatial resolution to demonstrate changes in renal perfusion. Depression of renal medullary perfusion after DP appears to be caused by the intrarenal effect of DP and may have clinical impact. PMID- 8724413 TI - Contrast enhancement with Gd-EOB-DTPA in MR imaging of hepatocellular carcinoma in mice: a comparison with superparamagnetic iron oxide. AB - The purpose of this study was to evaluate the ability of the new liver-specific magnetic resonance contrast agent gadolinium-ethoxybenzyl-diethylenetriamine penta-acetic acid (Gd-EOB-DTPA) to detect hepatocellular carcinoma (HCC). Seventeen mice with 66 chemically induced HCCs underwent magnetic resonance imaging with both Gd-EOB-DTPA (30 mumol/kg) and superparamagnetic iron oxide (SPIO; 10 mumol/kg). After enhancement, lesion-to-liver contrast-to-noise ratios (CNRs) of 47 detected HCCs increased negatively from 3.7 +/- 10.7 (mean +/- SD) to -55.1 +/- 25.8 with Gd-EOB-DTPA (P < .001) and increased positively from 10.4 +/- 10.4 to 26.1 +/- 16.3 with SPIO (P < .001). The improvement of CNR after administration of SPIO was less in smaller lesions (< 4 mm), whereas that after administration of Gd-EOB-DTPA was independent of lesion size. However, Gd-EOB DTPA positively enhanced four HCCs (8.5%), both highly differentiated (grade 1) and moderately differentiated (grade 2). Gd-EOB-DTPA allows the conspicuous detection of small HCCs; however, moderately differentiated HCCs occasionally may be positively enhanced. PMID- 8724412 TI - Rectal carcinoma: prospective comparison of conventional and gadopentetate dimeglumine enhanced fat-suppressed MR imaging. AB - The purpose of this study is to compare the usefulness of conventional MR imaging and gadopentetate dimeglumine enhanced fat-suppressed MR imaging for the depiction and staging of rectal carcinoma. Thirty-two patients were prospectively evaluated by MR imaging using a 1.5-T unit. Based on the results of a barium study and/or digital examination, a balloon catheter was inserted to the level of the lesion before examination. Both conventional T1- and T2-weighted images and gadopentetate dimeglumine enhanced fat-suppressed T1-weighted images were obtained for all patients. The kappa statistics were performed for the evaluation of interobserver agreement and the McNemar test was performed for the analysis of staging accuracy. When only T1- and T2-weighted images were used, 5 of 32 tumors were not detected and the extent of 18 of 32 tumors were unclear. However, when gadopentetate dimeglumine enhanced fat-suppressed imaging was added, 24 of 32 tumors were well defined and only one tumor was not detected. In determining the depth of invasion, the staging accuracy was 72% for conventional imaging and 68% for all images combined. There was no significant difference between with gadopentetate dimeglumine fat-suppressed imaging and conventional imaging (P > .05). Use of gadopentetate dimeglumine (fat-suppressed imaging) resulted in overestimation of muscular invasion, perirectal fat invasion, and adjacent organ invasion in 12 patients, whereas nine patients were overestimated without the use of gadopentetate dimeglumine. In the detection of metastatic lymph nodes, gadopentetate dimeglumine enhanced fat-suppressed imaging also was not useful. Tumor detection was excellent using gadopentetate dimeglumine enhanced fat suppressed images. However, the accuracy of staging was not improved by obtaining such images. PMID- 8724414 TI - Magnetic resonance imaging evaluation of the ascending aorta after graft inclusion surgery: comparison between an ultrafast contrast-enhanced MR sequence and conventional cine-MRI. AB - The aim of this study was to compare the value of two magnitude-based MR sequences, a contrast-enhanced ultrafast MR sequence and a cine MR sequence, for the detection of flow around the graft in the postoperative ascending thoracic aorta. Thirteen patients who underwent ascending aortic surgery for dissection (n = 11) and aneurysm (n = 2) were enrolled. They were referred to MRI for mediastinum enlargement on the chest x-ray or mediastinal symptoms. All patients had replacement of the diseased ascending aorta using a graft-inclusion technique (with wrapping of the native aorta over the graft). Delays between surgery and MRI ranged from 15 days to 8 years (mean: 10 months). All patients were examined using a T1-weighted spin-echo sequence, a magnitude-based cine gradient-echo sequence, and an ultrafast contrast-enhanced MR sequence. MR images of the cine and the contrast-enhanced sequences were retrospectively and independently analyzed by two qualified radiologists for evidence of flow or thrombus around the graft, including perigraft structure and aneurysmal formations. Evidence of flow was detected by both methods in all aneurysmal formations of the ascending aorta located at suture level (n = 6). Evidence of flow in the perigraft structure was found in seven cases by contrast-enhanced MR sequence and in only four cases by cine MR sequence. Contrast-enhanced ultrafast MR sequence could be proposed as a minimally invasive and fast modality for assessing the perigraft structure, in graft-inclusion surgical procedures of the ascending aorta, when noncontrast enhanced MR sequences do not exclude the possibility of flowing blood in this structure. PMID- 8724415 TI - Black blood dual phase turbo FLASH MR imaging of the heart. AB - An MR imaging scheme for dark blood cardiac images acquired simultaneously in end diastolic and end systolic phases, in breath-hold times, is presented. The sequence consists of a magnetization preparation period followed by two segmented k-space acquisitions. Image quality was investigated with respect to different sequence parameters (optimal values are indicated in brackets): (a) echo time (TE)/repetition time (TR)/ flip angle (FA) (3/6.2 msec/20 degrees); (b) number of lines/ segments in the acquisition window (11 lines/segment); (c) location of acquisition windows and inversion time; and (d) thickness of slice "reinverted" during preparation (1.53 times the acquisition slice thickness). The image quality of the basal slices at end systole was critically dependent on the last parameter. High quality short axis views of the heart, with good blood signal suppression, were acquired with the optimized sequence on four volunteers from apex to base at two phases in 10 breath-holds. PMID- 8724416 TI - Dynamic range extension of cine velocity measurements using motion-registered spatiotemporal phase unwrapping. AB - A motion-registered spatiotemporal phase-unwrapping method for extending the dynamic range of cine magnetic resonance phase velocity measurements is presented. The interframe cardiac movement is estimated from the magnitude image derived from the velocity encoded raw data, which ensures that the phase signal is unwrapped in the temporal direction with reference to pixels belonging to the same anatomic flow region. An extra step of spatial phase correction is then used to further eliminate any residual errors. The combination of spatial and temporal information for phase unwrapping provides a robust way of extending the dynamic range of cine velocity data in the presence of large phase aliasing errors. PMID- 8724417 TI - MR flow quantification using RACE: clinical application to the carotid arteries. AB - Real time MR flow quantification was performed with real time acquisition and evaluation of motion (RACE) in a rigid phantom under steady flow conditions and in the common carotid arteries of 43 subjects aged 24-78 years. Hemodynamic information included the intraluminal velocity distribution during the complete cardiac cycle, the distensibility of the arterial wall, and age-dependent changes of the flow curves. Systolic peak velocities of 51 +/- 33 cm/s and time-averaged volume flow rates of 4.3 +/- 2.0 ml/s were measured in healthy subjects. Flow rates below 3.0 ml/s and the observation of abnormal flow patterns indicated stenoses greater than 70% in the region of the bifurcation (sensitivity: 83.3%; specificity: 93.7%; accuracy: 71.4%). Improvements may be achieved from a combination with MR angiography, providing both functional and morphologic vascular information noninvasively within one observer-independent examination. MR imaging, therefore, has a strong potential for the diagnosis of critical stenoses in symptomatic patients. PMID- 8724418 TI - Physiologic motion phantom for MRI applications. AB - To address the need for a complex physiologic motion phantom for use in MR applications, such as the verification of techniques for measuring myocardial motion dynamics and motion insensitive pulse sequences, a computer-controlled motion phantom has been designed. The phantom, which consists of a deformable silicone gel annulus mounted on a translation stage, can undergo a range of bulk motions and deformations. Available motions include bulk rotation and translation, rotational shear, axial shear, and combinations of some or all of these motions. In this paper, the capability of the phantom to produce accurate constant and time-varying waveforms is demonstrated. In the current implementation, peak linear translation and rotation rates are 175 mm s-1 and 10 rad s-1, respectively. Cycle-to-cycle reproducibility is excellent, with variations of less than .003 radians over the period of hours while undergoing rotational shear. The phantom has been designed in a flexible fashion so that various test objects can be scanned while undergoing bulk translation and can be adapted to produce different deformations. PMID- 8724419 TI - Local histogram correction of MRI spatially dependent image pixel intensity nonuniformity. AB - We describe a computationally straightforward post-hoc statistical method of correcting spatially dependent image pixel intensity nonuniformity based on differences in local tissue intensity distributions. Pixel intensity domains for the various tissues of the composite image are identified and compared to the distributions of local samples. The nonuniformity correction is calculated as the difference of the local sample median from the composite sample median for the tissue class most represented by the sample. The median was chosen to reduce the effecters on determining the sample statistic and to allow a sample size small enough to accurately estimate the spatial variance of the image intensity nonuniformity. The method was designed for application to two-dimensional images. Simulations were used to estimate optimal conditions of local histogram kernel size and to test the accuracy of the method under known spatially dependent nonuniformities. The method was also applied to correct a phantom image and cerebral MRIs from 15 healthy subjects. Results show that the method accurately models simulated spatially dependent image intensity differences. Further analysis of clinical MR data showed that the variance of pixel intensities within the cerebral MRI slices and the variance of slice volumes within individuals were significantly reduced after nonuniformity correction. Improved brain cerebrospinal fluid segmentation was also obtained. The method significantly reduced the variance of slice volumes within individuals, whether it was applied to the native images or images edited to remove nonbrain tissues. This statistical method was well behaved under the assumptions and the images tested. The general utility of the method was not determined, but conditions for testing the method under a variety of imaging sequences is discussed. We believe that this algorithm can serve as a method for improving MR image segmentation for clinical and research applications. PMID- 8724420 TI - Multilevel wavelet-transform encoding in MRI. AB - This paper explores an alternative to standard Fourier MRI called wavelet transform encoding. Spatially selective radiofrequency (RF) pulses are used to excite slice profiles in the shape of the wavelet-basis functions of a discrete wavelet transform. Our implementation on a standard commercial whole body MRI system resolves one spatial dimension through wavelet encoding and orthogonal directions through conventional frequency encoding and rectangular-slice selection. The wavelet transform is described as a method for multiresolution analysis, and we show how a high resolution MR image can be constructed from lower resolution images, representing the approximation and detail structures of the object. A multiple level architecture of the wavelet reconstruction is described, thereby allowing wavelet-encoded images to be reconstructed through several modes. Multilevel wavelet-encoded images of a gel phantom are presented that show comparable image quality to Fourier-encoded images of similar signal-to noise ratio. However, a chemical-shift artifact is identified with this particular implementation. PMID- 8724421 TI - Determination of 1H relaxation times of water in human bone marrow by fat suppressed turbo spin echo in comparison to MR spectroscopic methods. AB - Relaxation times of water were measured in human vertebral bodies by a fat suppressed dual-echo turbo spin echo/turbo inversion recovery MRI sequence. Comparison was made with T1 and T2 values obtained by localized 1H-MR spectroscopy. The accuracy of the results and the diagnostic potential of the fast quantitative MRI technique were evaluated in 20 volunteers, 11 patients with osteoporosis, 6 patients with lymphoma, and 6 patients with bone marrow metastasis. No significant alterations of T1 and T2 relaxation times of water and fat were found in osteoporosis. With both methods, an increase in the T1 values of the water resonance by 16% was observed in lymphomas, which was highly significant (P < .001) in the MRS measurements, and an elevation by the same amount was obtained by the MRI sequence for the metastases (P = .040). A strong reduction of fat fraction was quantified by MRS in the tumorous cases. T2 of the water resonance increased by more than 30% (P < .003) in metastases. Water T2 values obtained by the MRI sequence showed systematic deviations from the MRS results, especially at short echo spacings. PMID- 8724422 TI - Investigation of MR decay rates in microphantom models of trabecular bone. AB - MR measurements of transverse relaxation time, T2*, in trabecular bone may provide both structural and density-related information for assessment of bone mineral status in osteoporosis. Using submillimeter scale glass phantoms as simplified models of trabecular bone, we have made a quantitative investigation of the dependence of T2* decay on modeled trabecular microstructure and MR image resolution. The experimental MR data are in excellent agreement with predictions from a computer simulation. Decreasing the modeled trabecular bone volume fraction, sigma, decreases the decay rate, as expected. However, if trabecular width and spacing are both increased without changing sigma, the decay rate is unchanged. The measured decay curves closely follow the predicted dependence on trabecular orientation. The decay rates are independent of image resolution, provided that the pixel dimensions are larger than the intertrabecular spacing. For smaller pixel sizes, the decay rate decreases with decreasing pixel size. PMID- 8724423 TI - An infrared device for monitoring the respiration of small rodents during magnetic resonance imaging. AB - A device that uses infrared reflectometry for monitoring the respiratory waveform of small rodents during MRI was developed. This system uses a photoplethysmograph coupled to the animal by a light pipe to detect movements as small as 1.0 mm. This device can also be used for monitoring larger animals or for controlling respiratory-compensated or respiratory-triggered MRI. PMID- 8724424 TI - The electrophysiological work of Carlo Matteucci. 1964. AB - In 1964, Giuseppe Moruzzi (1910-1986), a prominent neurophysiologist of worldwide fame with a scholarly interest in the history of science, published an extensive and insightful analysis of the work and personality of Carlo Matteucci, a 19th century Professor of Physics at the University of Pisa who had worked on "animal electricity" in the great tradition of Galvani. By discovering fundamental phenomena such as the demarcation potential and action current in muscle and nerve, Matteucci paved the way for the development of modern electrophysiology. Being written in Italian and having appeared in a not easily available journal of history of science, Moruzzi's essay has been lost to most members of the international neuroscience community. An English translation of it is published here with two aims: first, to make known to a large audience of neuroscientists a fascinating account of the origins of electrophysiology as seen through a perceptive interpretation of the successes and failures of a brilliant scientist and passionate man; and second, to pay tribute to the memory of Giuseppe Moruzzi and his everlasting contributions to learning and science on the occasion of the 10th anniversary of his death. PMID- 8724425 TI - Effects of lesions of amygdaloid nuclei and substantia nigra on aversive responses induced by electrical stimulation of the inferior colliculus. AB - Stimulation of the central nucleus of the inferior colliculus causes defensive behavior. In this work we examined the influence of lesions of brain structures involved in the expression of fear, such as periaqueductal gray matter, amygdala, and substantia nigra pars reticulata (SNpr), on these aversive responses. Thus, rats were implanted with an electrode in the central nucleus of the inferior colliculus, for the determination of the thresholds of alertness, freezing, and escape responses. Each rat also bore a cannula implanted in the periaqueductal, amygdala or Snpr for injection of the neurotoxin N-methyl-D-aspartate (8 micrograms/0.8 microliters). The data obtained show that lesion of the central nucleus of the amygdala increases the thresholds of aversive responses whereas lesion of the basolateral complex decreases the threshold of these responses. Lesion of the Snpr increased the aversive consequences of the electrical stimulation of the inferior colliculus whereas periaqueductal gray lesions, either dorsal or ventral regions, did not change these responses. From the evidences obtained in this work, it is suggested that the expression of the defensive behavior induced by activation of the neural substrates of the inferior colliculus does not seem to depend on the integrity of the periaqueductal gray. On the contrary, the basolateral complex inhibits and the central nucleus amplifies the aversive responses integrated in the inferior colliculus. Furthermore, SNpr seems also to be an important motor output for the defensive behavior induced by stimulation of the inferior colliculus, in agreement with what has been suggested for other brain structures implicated in the expression of fear. PMID- 8724426 TI - Central opiate modulation of growth hormone and insulin-like growth factor-I. AB - The effects of central administration of morphine-sulfate (MOR:80 micrograms) and morphine-6-glucuronide (M6G:1 microgram) on the growth hormone (GH)/insulin-like growth factor (IGF) system were assessed. MOR and M6G were injected intracerebroventricularly (ICV) in chronically catheterized 24 h fasted rats; time-matched control animals received H2O (5 microliters). MOR increased plasma GH concentrations 3-fold 2 h after i.c.v. injection, and transiently increased the plasma concentration and liver content of IGF-I (60% and 90%, respectively) 30 min after i.c.v injection. M6G did not produce any significant alterations in plasma GH and IGF-I levels at the time-points measured. Both MOR and M6G increased the concentration of IGF binding protein-1 (IGFBP-1) in plasma and liver 2 h after injection. However, MOR showed 2- to 2.5-fold greater effect than M6G in stimulating plasma and liver IGFBP-1. MOR and M6G produced similar increases in plasma epinephrine (5-fold), norepinephrine (3-fold) and corticosterone (1.5-fold). Neither opiate significantly altered circulating insulin levels. These findings suggest that opiate modulation of GH and IGF may be hormone-independent and centrally modulated. We speculate that differential affinities of MOR and M6G to the different opiate receptor subtypes might be responsible for their distinct effects on GH/IGF-I system. PMID- 8724427 TI - Gangliosides attenuate stress-induced changes on body weight, motor activity and on the behavioral response to 5-methoxy-N,N-dimethyltryptamine. AB - The major goal of this study was to evaluate the influence of gangliosides (GANG) treatment on the onset of adaptive changes and the sequelae induced by stress exposure. With this purpose, the behavioral response to 5-methoxy-N,N dimethyltryptamine (5-MeODMT, 5 mg/kg, IP) and motor activity were evaluated in rats previously submitted either to a single restraint session (2 h) or to a daily restraint event for 3 consecutive days, combined or not to GANG administration (30 mg/kg IP). GANG was always injected 2 h before stress exposure. In addition, differences in body weights were recorded throughout the experiments. A similar behavioral response after 5-MeODMT was observed between saline (SAL) and GANG unstressed rats. Exposure to one or three restraint sessions did not modify the behavioral response to 5-MeODMT, whereas the association of GANG and stress during 3 consecutive days enhanced forepaw treading and hindlimb abduction. SAL-treated animals submitted to a single or to three stressful stimuli showed reduced locomotion and rearing. The combination of GANG and stress for 3 days, but not after a unique association, reversed the decrease on motor activity induced by the aversive experience. The decrease of body weights produced by one or three stress sessions was recovered only in animals treated with GANG and stress for 3 days. These findings suggest that GANG may accelerate the onset of adaptive changes on 5-HT1 sites and attenuate certain sequelae induced by previous stress experience. PMID- 8724428 TI - Extracellular catechol and indole turnover in the nucleus of the solitary tract of spontaneously hypertensive and Wistar-Kyoto normotensive rats in response to drug-induced changes in arterial blood pressure. AB - Drug-induced alterations in arterial blood pressure are reflected in the extracellular fluid neurotransmitter levels of the nucleus of the solitary tract (NTS). Urethane-anesthetized spontaneously hypertensive rats (SHRs) and Wistar Kyoto normotensive (WKY) rats were used in this study. The extracellular neurochemical profile of the NTS was quantified using the in vivo microdialysis technique. In SHR, phenylephrine-induced hypertension produced no significant changes in the extracellular norepinephrine (NE) and dihydroxyphenylacetic acid concentrations, whereas a significant increase in the serotonin metabolite 5 hydroxyindoleacetic acid (5-HIAA) concentration was observed. Wistar normotensive rats, in response to phenylephrine-induced hypertension, showed a significant increase in extracellular NE and 5-HIAA concentrations. Hypotension produced by the intravenous infusion of nitroprusside failed to show significant changes in the extracellular neurotransmitters in both SHR and WKY rats. However, there was a significant increase in 5-HIAA concentration in SHRs during the rebound hypertension, which followed the nitroprusside-infused hypotension. No such change was observed in the case of the WKY rats. These results suggest the possible involvement of the serotonergic mechanisms of NTS in the regulation of normal arterial blood pressure in these two rat strains. PMID- 8724429 TI - The effects of prefrontal intracortical microinjections of an alpha-2 agonist, alpha-2 antagonist and lidocaine on the delayed alternation performance of aged rats. AB - A selective alpha-2 agonist medetomidine, a selective alpha-2 antagonist atipamezole and lidocaine were injected into the prefrontal cortex of aged rats that had been trained to perform a delayed alternation (DA) task in a T maze. Medetomidine at the dose of 0.01 microgram/microliter improved the memory task performance in four of five rats. The fifth rat improved its performance at the dose of 1.0 microgram/microliter. Atipamezole did not have any effect on the task performance. The effect of a systemic dose of medetomidine (3 micrograms/kg), which was previously shown to improve the task performance in old rats, was reversed by an intracortical injection of atipamezole. Lidocaine impaired the DA performance significantly. The results suggest that the beneficial effects of medetomidine on the working memory of old rats are mediated at least partly through the prefrontal cortex. PMID- 8724430 TI - Association of dopaminergic terminals and neurons releasing nitric oxide in the rat striatum: an electron microscopic study using NADPH-diaphorase histochemistry and tyrosine hydroxylase immunohistochemistry. AB - To examine synaptic input and association of terminals containing dopamine and other transmitters to rat striatal nitric oxide synthase-expressing neurons, an electron microscopic study using tyrosine hydroxylase (TH) immunohistochemistry combined with histochemistry for NADPH-diaphorase (NADPHd) was performed. NADPHd positive neurons had medium-sized cell bodies containing a highly invaginated nucleus and received relatively sparse synaptic input; 3.6% of boutons apposed to the NADPHd-positive neurons were TH-immunoreactive. Of these TH-immunoreactive boutons, two synaptic contacts showing symmetrical synaptic specializations were found on a cell body and a proximal dendrite of a NADPHd-positive neuron. Other nonsynaptic TH-immunoreactive boutons were occasionally associated with unlabeled terminals adjacent to the NADPHd-positive dendrites and also forming asymmetric synaptic contacts with unlabeled spinous or dendritic profiles. These results suggest that activity of the striatal neurons that release nitric oxide may be regulated by direct synaptic input from dopaminergic neurons and also suggest that the TH-immunoreactive terminals associated with the dendrites of nitric oxide synthase-expressing neurons provide the sites where nitric oxide influences dopamine release from neighboring terminals. PMID- 8724431 TI - State-dependent changes of brain endogenous opioids in mammalian hibernation. AB - Endogenous opioids belonging to three opioid families were measured in different states of the hibernation cycle in brain of the Columbian ground squirrels. Using high-performance liquid chromatography-EC detection, the hypothalamic and septal concentrations of met-enkephalin were found to be significantly higher (p < 0.05) in the hibernating state than that in the nonhibernating state. In contrast, met enkephalin content in the medulla decreased significantly during hibernation. Leuenkephalin content was only increased in the hypothalamus of hibernating animals. Using radioimmunoassay, dynorphin A-like immunoreactivity was observed to increase in the claustrum and striatum, whereas beta-endorphin-like peptides showed a significant increase in the hypothalamus during hibernation. It is evident that the changes in endogenous opioids in brain during hibernation are state dependent, type specific and region specific. These specific alterations of various endogenous opioids may imply their different roles in hibernation. PMID- 8724432 TI - Effects of neurotensin on neurons in the rat central amygdaloid nucleus in vitro. AB - The effects of neurotensin (NT) on neurons in the central amygdaloid nucleus (ACe) were investigated in rat brain slice preparations by adding the peptide to the perfusing medium. Of 115 ACe neurons, 69 cells (60%) showed excitatory responses and 10 cells (9%) showed inhibitory responses to application of NT. The excitatory response to NT was observed in a dose-dependent manner and the threshold concentration was approximately 3 x 10(-9) M. The excitatory effects of NT persisted under blockade of synaptic transmission. The NT fragment neurotensin 8-13 and the NT analogue neuromedin N showed effects similar to those of NT, whereas the NT fragment neurotensin 1-8 had no effect on ACe neurons. Of 43 neurons in the septal nucleus, 8 cells (19%) and 3 cells (7%) showed excitatory and inhibitory responses, respectively, to NT. The results suggest that NT exerts a potent excitatory effect on ACe neurons through a direct action on specific receptors, in which NT may play a role in amygdala-relevant functions. PMID- 8724433 TI - Neuronal activity in monkey visual areas V1, V2, V4 and TEO during fixation task. AB - We analyzed 577 neurons recorded from visual areas V1, V2, V4, and the inferotemporal area (TEO) of macaque monkeys, which performed a visual fixation task and a spot-off-on (blink) test during the fixation period. Among these neurons, 35% were defined as task-related cells, because they gave responses at the task-start, fixation, or task-end periods but were unresponsive to the spot blink, which was physically identical to these stimuli. Blink-responsive cells accounted for 29% and task-unresponsive cells for 30% of the neurons. The task related response was large and frequent in V4 (34%) and TEO (41%), but small and less frequent in V1 (31%) and V2 (27%). Other observations further demonstrated nonsensory activities in these areas: In some cells, response to the fixation spot was inhibitory, whereas light stimulation on the fovea was excitatory; some V1 and V2 cells had color-irrelevant responses, and some cells responded to the spot-off only when the monkey regarded it as a task-end cue. PMID- 8724434 TI - Effect of aqueous and nonaqueous fixatives on the quantitative estimation of collagen-proteoglycan interaction in tissue sections. AB - The present study was designed to assess the influence of aqueous and nonaqueous fixatives on the quantitative estimation of collagen-proteoglycan interaction in tissue sections. Tissues containing different collagen types and distinct sulfated proteoglycan classes were isolated from pig costal cartilage, human skin, and the inner muscular layer of dog small intestine and fixed using aqueous or nonaqueous methods. The results showed that the best fixation method was exposure to paraformaldehyde gas. When using aqueous fixatives, proteoglycans were lost to different degrees among the various tissues analyzed, reflecting differences in chemical properties of proteoglycan classes and/or in their interactions with other matrix components such as collagen. PMID- 8724435 TI - An improved method for preparing the chromosomes of pines and other gymnosperms. AB - A simple technic is described to produce well spread gymnosperm chromosomes. Root tip meristems are digested with a pectinase:cellulase mixture to produce a cell suspension which then is squashed to yield flat, well spread chromosome complements that can be stained or used for in situ hybridization. PMID- 8724436 TI - The influence of staining procedure on differential round cell analysis in stained smears of human semen. AB - Giemsa and Bryan-Leishman smear staining techniques have been quantitatively evaluated for their ability to determine round cells in human semen. Samples of fertile and vasectomy ejaculates were compared against counts obtained from semithin Araldite sections stained with toluidine blue. TEM studies and immunogold staining of the Pradite section permitted identification and quantitation of nucleated cell profiles. Differential counts from each of the three stains on the same set of semen samples were compared using regression analysis. Counts of seminiferous tubule elements from stain to stain correlated well (r > 0.9). Numerical analyses indicated, however, that leucocytes were commonly misidentified. The r values for neutrophils were less than 0.8 and as low as 0.55 for lymphocytes. These low correlations presumably were due to failures to distinguish between these cells and seminiferous tubule elements. PMID- 8724437 TI - The role of DNase and EDTA on DNA degradation in formaldehyde fixed tissues. AB - Degradation and extraction of high molecular weight DNA from formaldehyde fixed tissues suitable for gene analysis are presented. We previously reported that DNase might play an important role in the degradation of DNA extracted from formaldehyde fixed tissues (Tokuda et al. 1990). In the present study, DNase activity of the supernatant from rat tissues fixed in buffered formaldehyde at room temperature was negligible within 3 hr. Analysis of DNA extracted from reconstituted chromatin revealed that the degradation increased in the absence of DNase depending on the duration of the formaldehyde fixation. Furthermore, high molecular weight DNA could be extracted from tissues devoid of DNase activity fixed in buffered formaldehyde containing EDTA. These results demonstrated that DNA degradation was due mainly to a mechanism other than DNAse which was inhibited by EDTA. For clinical application, v-H-ras gene was successfully detected by Southern blotting from rat spleen tissues fixed in buffered formaldehyde especially at 4 C. Fixation at low temperature is useful for gene analysis. PMID- 8724438 TI - Highly selective acridine and ethidium staining of bacterial DNA and RNA. AB - The acridine dyes acridine orange (AO) and coriphosphine O (CPO) and ethidium bromide (EtBr) were used to stain bacterial digests after electrophoresis in native and denaturing (SDS) polyacrylamide gels and were shown to stain DNA and RNA preferentially over other subcellular components in the gels. Vegetative cell digests of Bacillus subtilis, Escherichia coli, Micrococcus luteus, and Staphylococcus aureus showed intense staining of DNA with AO and CPO near the top of the gel, but little or no staining of other cellular constituents. EtBr stained both DNA and RNA in the gels. Protein standards and non-nucleic acid cellular constituents stained faintly with high concentrations (> or = 100 microM) of AO, lower concentrations (13.9 microM) of CPO, and did not stain with 0.5 microgram/ml EtBr in denaturing gels. The complete set of cellular biochemicals was visualized by silver staining, while the protein subset was detected by Coomassie blue staining. The highest concentrations of AO (120 microM) and CPO (13.9 microM) were shown to detect purified DNA in gels with a sensitivity in the range of 25-50 ng per band. This work demonstrates the specificity of acridine and ethidium dyes for nucleic acids, while illustrating the level of non-nucleic acid-specific interactions with other cellular components by staining of electrophoretically separated cellular components in a gel matrix. PMID- 8724439 TI - Preparation of insect chromosomes for immunolabeling. AB - We describe a method for isolating chromosomes from testes of the desert locust, Schistocerca gregaria, and their subsequent incubation with antibodies directed against chromosomal proteins. The procedure involves hypotonic pretreatment of the germ cells, centrifugation onto coverslips in a cytocentrifuge and immunolabeling, while still unfixed, using a chromatin-stabilizing buffer. In the present case, an antibody specific for the acetylated isoforms of histone H4 was tested. After the antibody treatment, the preparations are fixed using formaldehyde, stained with a DNA-specific fluorescent dye and mounted. Analysis of the preparations revealed good preservation of chromosome structure in prophase spermatogonia and late prophase I spermatocytes. Fully condensed chromosomes were not observed and are probably lost during preparation. The bright fluorescence of the autosomes indicates that the reaction between the antibody against acetylated histone H4 and its chromosomal antigen is not impeded. In contrast, the X univalent remained unstained with the exception of a small terminal band. Thus, cytospin preparations of locust germ cells allow high resolution immunolabeling with antibodies against chromosome-associated proteins. PMID- 8724440 TI - The Gram stain after more than a century. AB - The Gram stain, the most important stain in microbiology, was described more than a century ago. Only within the past decade, however, has an understanding of its mechanism emerged. It now seems clear that the cell wall of Gram-positive microorganisms is responsible for retention of a crystal violet:iodine complex. In Gram-negative cells, the staining procedures damage the cell surface resulting in loss of dye complexes. Gram-positive microorganisms require a relatively thick cell wall, irrespective of composition, to retain the dye. Therefore, Gram stainability is a function of the cell wall and is not related to chemistry of cell constituents. This review provides a chronology of the Gram stain and discusses its recently discovered mechanism. PMID- 8724441 TI - Detection of S-phase cells in smear cytology using in vitro bromodeoxyuridine labeling. AB - A technique is described for rapid detection of S-phase cells of tumor tissues in smear specimens using bromodeoxyuridine (BrdU) immunostaining. Mouse NR-S1 tumors and human tumor specimens were prepared for smear cytology after incubation in RPMI 1640 culture medium containing 200 microM BrdU at 37 degrees C under 3 atm for 1 hr. Samples were fixed in 70% ethanol for 30 min and used immediately or air dried for 30 min. Samples were then denatured in either 4 N HCl or 0.07 N NaOH to prepare partially single-stranded DNA. Fixation with air drying for 30 min followed by 30 min in 70% ethanol and 1 min denaturation with 0.07 N NaOH resulted in satisfactory staining quality. Cultured tumor specimens were processed for routine paraffin sections after smears were made for cytology. The labeling indices of the smear specimens and of the paraffin sections gave similar results. This technique should be useful in evaluating the cell proliferative potential of tumor tissue in smear cytology without processing paraffin sections. PMID- 8724442 TI - A new method for detecting nucleolus organizer regions in fish chromosomes using denaturation and propidium iodide staining. AB - A rapid method for detecting nucleolus organizer regions (NORs) in fish chromosomes based on thermal denaturation and staining with propidium iodide is described. Under epifluorescence, the NORs of 15 fish species from six families could be detected. This protocol differentiates constitutive heterochromatin in mammalian and avian chromosomes, and in some cases, heterochromatic blocks in fish chromosomes. The staining of NORs of fish chromosomes with propidium iodide following denaturation with formalin is likely the result of differential denaturation of the rDNA due to the thermal characteristics of AT- and GC-rich domains of the rDNA cistron. This technique provides a new useful marker for descriptive fish cytogenetic studies. PMID- 8724443 TI - A comparison of the acute behavioral effects of flunitrazepam and triazolam in healthy volunteers. AB - Flunitrazepam is an hypnotic benzodiazepine marketed in different European countries. Epidemiological studies have shown that it is frequently abused by opioid addicts. In a survey, "liking" scores for flunitrazepam in methadone maintenance patients were higher than ratings for other benzodiazepines. A double blind, placebo controlled, crossover clinical trial was conducted to assess the acute behavioral effects of flunitrazepam (0.50 and 2 mg) and triazolam (0.25 and 0.50 mg) in healthy male volunteers. Drug effects on physiological measures, psychomotor performance, and subjective rating scales, including specific questionnaires to evaluate abuse liability (e.g., ARCI or "liking" scores), were assessed before and 6 h after drug administration. Flunitrazepam 2 mg produced the most intense disruptive effects on all the performance tasks, triazolam 0.50 impaired performance except balance. All study drugs at all doses produced sedation symptoms in all or part of the subjective effects questionnaires. Only flunitrazepam 2 mg induced significative increases in some of the scales ("liking", "good effects", "high") that could be related to a possible abuse potential. The results seem to indicate that flunitrazepam, when administered to healthy subjects, produces some pleasurable subjective feelings, that could indicate a higher abuse liability of this drug as compared with other benzodiazepines. PMID- 8724444 TI - Self-administration of the dopamine D3 agonist 7-OH-DPAT in rhesus monkeys is modified by prior cocaine exposure. AB - Cocaine self-administration was compared with responding maintained by the dopamine D3 agonist 7-hydroxy-N,N-di-n-propyl-2-aminotetralin (7-OH-DPAT) in rhesus monkeys. In the first experiment, monkeys (n = 3) with an extensive cocaine history responded under a fixed-interval (FI) 5-min schedule of IV cocaine (0.03 mg/kg per injection) presentation, during daily 4-h sessions. When responding was stable, dose-response curves were determined for cocaine (0.01-0.3 mg/kg per injection) and 7-OH-DPAT (0.001-0.1 mg/kg per injection); each dose was available for at least five consecutive sessions. When substituted for the baseline dose of cocaine, other doses of cocaine and 7-OH-DPAT maintained rates higher than responding maintained by saline injections, in all monkeys. 7-OH-DPAT maintained response rates equal to or higher than rates of cocaine-maintained responding in all monkeys. In a second experiment, acquisition of 7-OH-DPAT self administration was evaluated in a group of cocaine-naive monkeys (n = 3). Various doses of 7-OH-DPAT (0.003-0.03 mg/kg per injection) were available under an FI 15 s schedule during daily 4-h sessions. After 10-13 sessions, 7-OH-DPAT self administration could not be trained in any cocaine-naive monkey. When cocaine was made available to these monkeys, responding was reliably maintained within one to four sessions and the schedule was gradually increased to FI 5-min. After stable responding under an FI 5-min schedule of 0.03 mg/kg per injection cocaine presentation, 7-OH-DPAT (0.01 mg/kg per injection) was again made available to two of the monkeys, and maintained responding at rates higher than saline. To determine better whether a history of responding maintained by another reinforcer would result in high rates of 7-OH-DPAT self-administration, two cocaine-naive monkeys were trained to respond under an FI 5-min schedule of food presentation. Substituting 7-OH-DPAT (0.003-0.03 mg/kg per injection) for food resulted in very low rates of responding. Taken together, these results suggest that despite comparable reinforcing effects in cocaine-substitution studies, 7-OH-DPAT and cocaine differ in their rate of acquisition, perhaps indicating a lower abuse liability for 7-OH-DPAT. PMID- 8724445 TI - Dizocilpine prevents the development of tolerance to ethanol-induced error on a circular maze test. AB - Dizocilpine [(+)MK-801] and ketamine, in doses that disrupt learning and memory, also prevent the development of tolerance to the motor impairing effects of ethanol (EtOH). However, dizocilpine itself affects motor behavior. In order to separate the possible influence of these two effects on the development of tolerance to EtOH, food-reinforced performance on a circular maze test was used in two different experiments. EtOH alone (1.2 g/kg) tended to increase the error score and reduce number of runs per trial, running speed, and total distance run, but on chronic administration of EtOH, tolerance developed progressively to all these effects. Dizocilpine also increased the error score, but had a biphasic effect on measures of running: low and intermediate doses (0.009 and 0.075 mg/kg, IP) increased running distance, whereas a high dose (0.15 mg/kg) decreased running speed and distance. When combined with EtOH, dizocilpine tended to overcome the effect of EtOH on running activity, but not on error score. Chronically, dizocilpine (0.075 and 0.15 mg/kg) prevented the development of tolerance to the effect of EtOH on error score, even though the lower dose of dizocilpine permitted tolerance to the effects of EtOH on running. These results suggest that NMDA receptor antagonists selectively inhibit tolerance to cognitive effects of ethanol even when the antagonists do not affect motor performance. PMID- 8724446 TI - Repeated 7-OH-DPAT treatments: behavioral sensitization, dopamine synthesis and subsequent sensitivity to apomorphine and cocaine. AB - Male Wistar rats (250-350 g) were injected (SC) daily with the putative selective dopamine D3 receptor agonist, 7-OH-DPAT (0.01, 0.10, or 1.0 g/kg) or vehicle for 10 days. Fifteen minutes after each injection, the rats were tested for locomotor activity in photocell arenas for 20 min or 2 h. In two experiments, following this subchronic treatment, all rats received a challenge injection of apomorphine (1.0 mg/kg, SC), or cocaine (10 mg/kg, IP) on day 11, and were tested for locomotor activity. In a third experiment, dopamine synthesis in striatal and mesolimbic (nucleus accumbens-olfactory turbercle) tissue was assessed following acute or chronic 7-OH-DPAT treatments by measuring the accumulation of dihydroxyphenylalanine (DOPA) after treatment with a DOPA decarboxylase inhibitor. Major findings were as follows: a) acute 7-OH-DPAT treatment produced a dose-dependent decrease in locomotor activity; b) when tested for 2 h, the 1.0 mg/kg dose of 7-OH-DPAT produced a progressively greater increase in activity across the 10 test days (i.e., behavioral sensitization); c) subchronic treatment with 7-OH-DPAT did not result in cross-sensitization to either apomorphine or cocaine; d) acute treatment with the 1.0 mg/kg dose of 7-OH-DPAT significantly decreased dopamine synthesis in both striatal and mesolimbic regions; and e) chronic 7-OH-DPAT treatments did not affect basal dopamine synthesis in either brain region. Although the behavioral effects of 7-OH-DPAT were similar to the reported effects of the D2/D3 dopamine agonist quinpirole, the effects of repeated 7-OH-DPAT treatments differed from those of quinpirole in terms of cross sensitization and basal dopamine synthesis. These results suggest that locomotor inhibition produced by low doses of 7-OH-DPAT is not related to dopamine autoreceptor stimulation, and the development of behavioral sensitization to high doses of 7-OH-DPAT is not due to the development of dopamine autoreceptor subsensitivity. PMID- 8724447 TI - Time-dependent exacerbation of amphetamine-induced taste aversions following exposure to footshock. AB - Previous studies have shown that stressors attenuate LiCI-induced conditioned taste aversions (CTA) but not morphine-induced CTA. The current studies examined the effects of footshock on the acquisition and extinction of amphetamine-induced CTA. Experiment 1 demonstrated that exposure to 30 footshocks between saccharin consumption and amphetamine injections did not alter either the acquisition or the extinction of amphetamine-CTA. Experiment 2 demonstrated that exposure to the same shock parameters 2 and 4 days before saccharin-amphetamine pairing increased the magnitude of amphetamine-CTA after one saccharin-amphetamine pairing and delayed the recovery from the CTA. Experiment 2 also demonstrated that footshock increased the initial neophobic response to novel saccharin but did not alter subsequent saccharin consumption among saline-injected animals. These results indicate that stress-induced facilitation of amphetamine CTA are time-dependent and contrast with reports that stressors attenuate LiCI CTA. They also add support to the contention that CTAs induced by self-administered drugs like amphetamine are qualitatively different from CTAs induced by toxic substances like LiCL. PMID- 8724448 TI - Age-related decline in central cholinergic function demonstrated with scopolamine. AB - Scopolamine hydrobromide was administered intravenously to 23 normal subjects (40 89 years) in doses of 0.1 mg, 0.25 mg, and 0.5 mg, in a double-blind. Placebo controlled, random-order fashion. The effects of scopolamine, as compared to placebo, were assessed using a comprehensive cognitive test battery, as well as behavioral and physiological measures. Scopolamine produced the expected dose dependent impairments in most of the cognitive functions assessed. Behavioral and physiological measures were also affected, but only minimally. More importantly, there was a significant overall correlation between age and scopolamine-impaired performances on psychomotor speed, short-term recall, visual tracking speed, visuo-motor coordination, and sequencing ability. There was, however, some inter individual variability in this phenomenon. The results provide further evidence that cholinergically mediated cognitive functions show an increased sensitivity to scopolamine with age, albeit with heterogeneity that bears further investigation. PMID- 8724449 TI - Neurotensin attenuates the reduction in alcohol drinking produced by angiotensin II. AB - Neurotensin enhances some of the behavioral effects of alcohol including motor impairment, narcosis, hypothermia and also interacts with some of the physiological actions of angiotensin (ANG) II including aldosterone release and increased blood pressure. ANG II injections also produce a dose-dependent antagonist reversible reduction in alcohol drinking. The present study is the first to examine the interaction between neurotensin and angiotensin in the behavioral context of oral alcohol self-administration. Adult male Wistar rats acquired alcohol drinking (6% w/v) using the limited access procedure which makes alcohol available for 40 min every day. When intake stabilized ANG II (400 micrograms/kg per day) or vehicle were administered subcutaneously (SC) just prior to alcohol availability but only the group receiving ANG II showed a marked reduction in alcohol intake. Following this the groups were pretreated sc with either vehicle or ascending doses of neurotensin (5, 10, 20 micrograms/kg) followed by either ANG II or vehicle. Control groups received either two vehicle injections or vehicle and neurotensin injection. Neurotensin alone did not affect alcohol intake at any of the doses tested but did attenuate, in a dose-dependent fashion, the reduction in alcohol intake produced by ANG II. These results demonstrate neurotensin's ability to alter the behavioral effect of ANG II on alcohol intake. PMID- 8724450 TI - Ovarian steroids modify the behavioral and neurochemical responses of the central benzodiazepine receptor. AB - The effect of ovarian steroids on the benzodiazepine receptor was assessed in the elevated plus-maze and, after restraint stress, in benzodiazepine receptor binding assays. Vehicle-treated proestrous rats displayed anxiolytic behavior, relative to diestrus or estrous rats. Anxiolytic behavior was observed after 1 or 2 mg/kg diazepam in diestrus and estrus. However, whereas 4 mg/kg increased open arm exploration in diestrus, a decrease in the same measure was found at estrus. At proestrus, a decrease in anxiolytic behavior was observed after 2 and 4 mg/kg. In ovariectomized vehicle-treated rats, restraint stress increased NaCl-induced potentiation of 3H-flunitrazepam binding in cortical and cerebellar, but not in hippocampal membranes. Estradiol benzoate (2 micrograms) prevented the potentiation of flunitrazepam binding by NaCl in nonstressed and stressed animals, whereas progesterone (0.5 mg) increased the NaCl-induced potentiation of flunitrazepam binding in both nonstressed and stressed animals. Combined estradiol benzoate and progesterone treatment produced effects that were intermediate to those seen after injection of either steroid alone. The potentiation of flunitrazepam binding by NaCl observed in vehicle-treated stressed or progesterone-treated nonstressed animals was mimicked in vitro by addition to reaction test tubes of the neuroactive metabolite of progesterone, 3 alpha-hydroxy-5 alpha-pregnan-20-one (allopregnanolone). These results point to a significant role of ovarian hormones in modifying the stress response of the benzodiazepine receptor. PMID- 8724451 TI - Fawn-hooded rats show enhanced active behaviour in the forced swimming test, with no evidence for pituitary-adrenal axis hyperactivity. AB - Fawn-hooded (FH) rats have been reported to have high basal corticosterone levels that can be normalized by antidepressant administration. In the present work, some behavioural and endocrine aspects were compared in FH and Sprague-Dawley (SD) rats. No interstrain differences in basal corticosterone levels or response to acute tail-cut stress were observed either in the morning or in the evening. Relative adrenal weight was lower in FH than SD, and relative thymus weight was, accordingly, higher in FH than SD rats. FH rats were hyperactive in the holeboard and showed behaviour similar to that of SD in the plus-maze. In contrast, FH rats showed greater levels of active behaviour (struggling) and lower levels of immobility than SD rats in the forced swimming test (FST). After desipramine (DMI) administration the differences between the two strains in the FST were more marked than in vehicle-treated animals. These results indicate that FH rats show no signs of pituitary-adrenal (PA) hyperactivity or depression-like behaviour, and therefore they are not an appropriate animal model of depression. PMID- 8724452 TI - Effect of the 5-HT1A agonist, 8-OH-DPAT on instrumental performance in rats. AB - These experiments assessed whether reported increases in food consumption and food-reinforced instrumental performance in undeprived rats by the 5-HT1A agonist 8-hydroxy-2(di-n-propylamino)tetralin (8-OH-DPAT) are due to an increment in the incentive value of foods. Against this hypothesis, we found that when undeprived rats were trained to lever press for the food pellets and then allowed to consume the pellets under 8-OH-DPAT, this reexposure decreased subsequent instrumental extinction performance regardless of test drug condition relative to reexposure under vehicle. Although both food consumption and reinforced lever press performance were incremented, 8-OH-DPAT was found generally to reduce instrumental extinction performance and lever pressing during a period when the reinforcer was delivered non-contingently. Rats injected with 8-OH-DPAT were, however, more sensitive to delay of reinforcement, and increased their lever press performance at a 3-s delay but decreased performance at 6-s and 12-s delays relative to animals injected with vehicle. These results are consistent with the hypothesis that 8-OH-DPAT modifies arousal processes in a manner similar to mild stress, thereby acting both to elevate rewarded instrumental performance and to increase sensitivity to the effects of non-reward. PMID- 8724453 TI - Somatodendritic 5-HT1A receptors are critically involved in the anxiolytic effects of 8-OH-DPAT. AB - In the rat shock-induced ultrasonic vocalization test, the anxiolytic effects of the 5-HT1A receptor agonist 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT) obtained after systemic (IP) and intracerebral injection into the dorsal raphe nucleus (DRN) were selectively abolished by pretreatment with the 5-HT1A receptor antagonist WAY-100635 [N-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]-N-(2 pyridinyl) cyclo-hexane-carboxamide trihydrochloride]. This blockade was demonstrated both after systemic and DRN application of WAY-100635. Therefore, it is concluded that the anxiolytic effects of 8-OH-DPAT are mediated by activation of somatodendritic 5-HT1A receptors. PMID- 8724454 TI - Dose-dependent aversive and rewarding effects of amphetamine as revealed by a new place conditioning apparatus. AB - Amphetamine-induced place conditioning was evaluated in mice using a newly designed apparatus. It was demonstrated that this apparatus provides a neutral set of cues devoid of rewarding or aversive properties and can reveal place preference or aversion after pairings with drugs (amphetamine and morphine for preference and naloxone for aversion) known to produce such effects. Moreover, repeated pairings of environmental cues with either 2 or 3 mg/kg d-amphetamine resulted in significant conditioned place preference on a drug free test, whilst repeated pairings with a lower dose of the drug (1 mg/kg) resulted in significant conditioned place aversion. Finally, a small number of mice showed opposite responses in comparison with group means at low as well as at high doses of amphetamine. These results suggest that amphetamine may promote conditioned place preference or avoidance depending on dosage and individual susceptibility. PMID- 8724455 TI - Carbohydrate histochemistry of vertebrate taste organs. PMID- 8724456 TI - Repetitive strain injuries and cumulative trauma disorders. PMID- 8724457 TI - Outcome assessment for carpal tunnel surgery: the relative responsiveness of generic, arthritis-specific, disease-specific, and physical examination measures. AB - Clinical evaluation of outcome after treatment of carpal tunnel syndrome has not been standardized. To assess the value of various clinical and questionnaire measures for the assessment of outcome after carpal tunnel surgery, we surveyed 22 patients 1 day before and 3 months after carpal tunnel release with the following measures: the Medical Outcomes Study 36-item short form health survey, the Arthritis Impact Measurement Scale, the Brigham and Women's Hospital carpal tunnel questionnaire, wrist range of motion, power pinch grip strength, pressure sensibility, and dexterity. Significant changes, all in the direction of improved health status postoperatively, were noted in the following scales or measures: the Arthritis Impact Measurement Scale pain, satisfaction, health perception, arthritis impact, and symptom scales; the Brigham and Women's Hospital symptom and function scales; the short form health survey's physical role, emotional role, and bodily pain scales; and the measurement of dexterity. In this study, standardized questionnaires were more sensitive to the clinical change produced by carpal tunnel surgery than many commonly performed physical measures of outcome. The condition-specific questionnaire was more sensitive to change than were more generic questionnaires. PMID- 8724458 TI - Outcome of reoperation for carpal tunnel syndrome. AB - One hundred thirty-one patients with reoperation for carpal tunnel syndrome were followed for a mean of 11 years. Reoperation failed in 15 patients, necessitating a third operation. Satisfaction, symptom severity, and functional status scores were assessed with a standardized questionnaire in the other 116 patients. Patients with normal findings on preoperative nerve conduction studies, those who filed for compensation, and those who had pain in the distribution of the ulnar nerve had significantly worse results. Those with abnormal findings on nerve conduction studies who had not filed for compensation had the best symptom and function scores and satisfaction at latest follow-up examination; those with normal findings on nerve conduction studies who had filed for compensation had the poorest outcome. Although most patients were satisfied with the overall outcome, many reported residual symptoms; in addition to the 15 patients who required a third operation, 22 patients were dissatisfied with the final result. PMID- 8724459 TI - Intergroup comparisons of carpal tunnel dimensions. AB - Although it is a common disease entity worldwide, idiopathic carpal tunnel syndrome is rarely seen in black South Africans. This study compares anatomic dimensions of the carpal canal in black and white population groups, using computed tomography. No significant anatomic differences were found between the groups tested; thus, other explanations need to be sought to explain the difference in occurrence of carpal tunnel syndrome in these groups. A parameter, the carpal canal index, was defined and may prove useful in future investigations of this region. PMID- 8724460 TI - Acute carpal tunnel syndrome caused by hydroxyapatite crystals: a case report. PMID- 8724461 TI - Compression of the median nerve proximal to the carpal tunnel in scleroderma. PMID- 8724462 TI - The hypothenar adductor muscle: an anomalous intrinsic muscle compressing the ulnar nerve. PMID- 8724463 TI - Superficial radial nerve compression at the elbow due to an accessory brachioradialis muscle: a case report. PMID- 8724464 TI - Innervated radial thenar flap for sensory reconstruction of fingers. AB - A radial thenar flap, which is supplied by the superficial palmar branch of the radial artery and innervated by the superficial branch of the radial nerve, was used in four patients for sensory reconstruction of the fingers. Two patients had degloving injuries of the hand and two had amputation injuries of the thumb. Three of the cases underwent free-flap transfers, and one case underwent a reverse-flow island flap. The size of these flaps was about 3 x 5 cm except in one case, where a 15 x 5 cm flap combined with a radial forearm flap was used. Sensory reinnervation was excellent and donor site morbidity was minimal in all four cases. An innervated radial thenar flap from the ipsilateral hand offers an alternative choice to reconstruct large palmar defects of the fingers. PMID- 8724465 TI - Restoration of sensibility in the hand after complete brachial plexus injury. AB - Twenty-one patients with complete brachial plexus palsy due to the avulsion of multiple cervical nerve roots underwent motor and sensory reconstruction. Of these, 15 patients who had been followed for at least 2 years were included in the present study. Sensory reconstruction was performed by nerve graft or nerve crossing to the median nerve. The donor nerves included the supraclavicular nerve in 10 patients, the intercostal nerve in 3, and the C5 nerve root in the first 2 patients. Limited sensibility in the median nerve distribution of the hand was restored in 12 patients, and the results were classified as S2+ in 2, S2 in 4, S1 in 6, and S0 in 3, according to the S0 to S4 grading system. No moving two-point discrimination was restored in any case. While the best recovery obtained was only at the S2+ level, even this limited sensibility was useful for otherwise completely anesthetic patients. PMID- 8724466 TI - Spinal accessory neurotization for restoration of elbow flexion in avulsion injuries of the brachial plexus. AB - Traumatic root avulsion brachial plexus injuries in 216 patients were treated with spinal accessory-musculocutaneous neurotization to restore elbow flexion. The average postoperative follow-up period was 6 years, with a minimum of 2 years for all patients. The percentage of satisfactory biceps recovery (MRC III or better) was 72.5%. The average interval between the operation and MRC III motor recovery was 17 months. The percentage of poor results increased from 25.5% to 62.5% in patients who underwent operation later than 9 months after injury. This method of neurotization produces a result comparable with, if not better than, the results of other types of neurotization in restoration of elbow flexion. PMID- 8724467 TI - Neurologic injury from axillary block anesthesia. AB - Three cases of severe permanent neurologic injury following axillary block anesthesia from a single hand surgery practice are presented, along with the results of a survey regarding complications of axillary block anesthesia sent to the membership of the American Society for Surgery of the Hand. This survey resulted in 800 replies. Of respondents, 171 (21%) had seen a major neurologic complication at some time in their practice. Minor, transient complications were reported by 521 respondents (65%). Only 232 respondents (29%) reported no major or minor neurologic complications in their practice. These cases and the results of this survey suggest that serious permanent neurologic injury following axillary block anesthesia, although rare, may be more frequent than previously believed. Transient neurologic problems following axillary block are common but generally do not lead to long-term problems. Permanent neurologic injury following axillary block anesthesia is a devastating complication and should be carefully considered when determining the preferred anesthesia for an individual patient. PMID- 8724468 TI - Which setting of the dynamometer provides maximal grip strength? AB - The purpose of this study was to determine which setting of the clinical dynamometer produced maximal grip strength and to correlate that setting with characteristics of the individual. Using a computerized isometric dynamometer, the maximal grip strength was determined in 288 normal asymptomatic hands in 4- to 78-year-old individuals at five dynamometer size settings. The majority of hands (256, or 89%) had a maximal grip strength at setting II. Mean body weight and mean height were lower in the group that had a maximal strength at or equal to setting I than in those individuals with maximal setting II and above. However, no clear significant correlation between body mass or hand size and maximal setting was evident. Those with a maximal grip at setting III or IV did not have larger hands or other features that differed from those with maximal settings at II. If measured at setting II only, maximal grip strength would have been underestimated in about 10% of individuals, but the error was small, ranging from 2% to 24%. Since no feature was identified that predicted maximal grip strength setting, it is recommended that grip strength of all patients should be routinely measured at setting II, irrespective of age, weight, or hand dimensions. PMID- 8724469 TI - Sensitivity of the Jamar Dynamometer in detecting submaximal grip effort. AB - Twenty-two people with no extremity disability were tested in a standard fashion using the Jamar Dynamometer to establish their maximum grip strength. Each participant was asked to grip first right-handed then left-handed three consecutive times. The directions were reexplained so that each participant would give a consistent, less than optimal effort; three trials right and three trials left were recorded. The standard deviations of each set of these trials were calculated for both right and left hands. These standard deviations were then tabulated as scores for 44 trials of 22 patients, both hands, for maximal and submaximal efforts. These scores were then compared, maximal versus submaximal, using a paired t-test. We found no statistical difference in the two groups in comparing the variability of results. Therefore, the current protocol for Jamar testing can allow a patient to make a consistently submaximal effort, resulting in a false apparent loss of grip strength. PMID- 8724470 TI - Clinical results of treatment of triangular fibrocartilage complex tears by arthroscopic debridement. AB - In order to determine indications for arthroscopic debridement and the management of triangular fibrocartilage complex (TFCC) tears, we reviewed 16 wrists retrospectively. The mean patient age was 30 years, with a range of 20 to 53 years. The follow-up period averaged 35 months. Two groups were identified: post traumatic tears (n = 11) and degenerative tears (n = 5). The results of arthroscopic debridement were compared and analyzed based on the preoperative and postoperative evaluation of pain, range of motion, grip strength, return to work, patient acceptance, and complications. Failures were further evaluated to determine identifiable lesions or anatomic defects associated with poor results. Patients with positive ulnar variance and lunotriquentral interosseous ligament tears had a poor clinical outcome. Good results correlated with grip strength; all patients with post-traumatic TFCC tears had excellent results, while those with degenerative TFCC tears did poorly. PMID- 8724471 TI - Arthroscopic management of partial scapholunate and lunotriquetral injuries of the wrist. AB - Outcome was studied in 14 patients with chronic mechanical (crepitant) wrist pain (symptomatic for more than 6 months) who had undergone arthroscopic debridement and early mobilization for the treatment of isolated partial scapholunate and lunotriquetral ligament injuries. Fourteen patients were available for a minimum 2-year follow-up interview, physical examination, and x-ray film evaluation. Eleven patients had complete relief of symptoms and had returned to work within 7 weeks of surgery. The other three (all workers' compensation cases) reported occasional mild wrist pain; two had returned to work a mean of 6.5 weeks after surgery, and the third did not return for 1 year. None of the patients had a statistically significant loss of grip or pinch strength. X-ray films and dynamic examination under fluoroscopy demonstrated no abnormalities. Thirteen of the 14 patients were highly satisfied (although 2 patients still had occasional mild wrist pain). Arthroscopic debridement of partial intrinsic ligament injuries provided excellent relief of painful crepitant symptoms and allowed for early return of wrist function in 13 of 14 cases. PMID- 8724472 TI - Classification and treatment of ulnar styloid nonunion. AB - Symptomatic nonunion of the ulnar styloid is an uncommon problem that is usually best treated by simple excision of the ulnar styloid fragment. Two types of nonunion of the ulnar styloid are described here on an anatomic basis, and their treatment differs. Type 1 is defined as a nonunion associated with a stable distal radioulnar joint. Type 2 is defined as a nonunion associated with subluxation of the distal radioulnar joint. The postoperative follow-up period for the two types ranged from 4 months to 13 years, with a mean of 5 years 2 months. Eleven type 1 wrists were treated with excision of the fragment, and all patients had satisfactory relief of pain. Nine type 2 wrists required restoration of the anatomy of the traingular fibrocartilage complex. Three of these had large fragments that were treated by open reduction and internal fixation. All three patients were completely relieved of their discomfort. Six other patients underwent excision of the fragment and repair of the triangular fibrocartilage complex to the distal ulna. This group had four excellent, one good, and one fair result. If the distal radioulnar joint is stable on presentation or if its stability is restored, then long-term relief of pain from ulnar styloid nonunion is achieved by treatment of the nonunion. PMID- 8724473 TI - Solitary osteochondroma of the scaphoid: a case report. PMID- 8724474 TI - Osteoblastoma of the scaphoid--long-term results following curettage: a case report. PMID- 8724475 TI - Isolated injuries to the dorsoradial capsule of the thumb metacarpophalangeal joint. AB - Isolated injuries to the dorsoradial capsule of the thumb metacarpophalangeal joint are rarely reported. We report 11 patients who had a tear in the dorsoradial capsule of the thumb metacarpophalangeal joint without an associated collateral ligament injury. Seven of the injuries were confirmed at surgical exploration and repaired by imbrication/direct repair. Six of these seven patients were able to return to unrestricted activities without requiring analgesics. The surgical patients had increased baseline metacarpophalangeal joint flexion compared to "average," which may predispose them to isolated capsular injuries. PMID- 8724476 TI - Combined extensor pollicis brevis and radial collateral ligament injury: three case reports. PMID- 8724477 TI - Distal interphalangeal joint arthrodesis comparing tension-band wire and Herbert screw: a biomechanical and dimensional analysis. AB - Thirty cadaveric distal interphalangeal joints (15 male and 15 female joints) were prepared with either a Herbert screw or a tension-band wire technique to simulate an arthrodesis. To elucidate mechanical differences between these constructs, the strength of the specimens was determined for three-point anteroposterior and lateral bending and for axial torsion. The Herbert screw demonstrated significantly greater anteroposterior bending strength and greater torsional rigidity when compared to the tension-band wire technique. For dimensional analysis, the height and width of each distal phalanx was measured prior to fixation, 4 mm from the distal tip of the bone (the region that must accommodate the large-diameter threads of the Herbert screw). Results indicated that the mean height of the distal phalanx (3.55 mm) is smaller than the diameter of the screw (3.90 mm). Fracture or thread penetration at the tip of the distal phalanx during screw placement occurred in 25 of the specimens overall and in all the female phalanges, often resulting in stretching or violation of the nail bed. Despite fracture or screw penetration, the Herbert screw appears to offer additional strength that may be clinically important for joint arthrodesis. PMID- 8724478 TI - Efficiency of the flexor tendon pulley system in human cadaver hands. AB - The efficiency of the flexor tendon system was examined in a human cadaver model. Pulleys were randomly sectioned, and the results were evaluated on the basis of the tendon excursion, force generated at the fingertip, and the work (force multiplied by distance) involved, as compared to the intact pulley system. When a single minor pulley (A1 or A5) was cut, there was no statistical difference in work efficiency or excursion efficiency from controls. Cutting all minor pulleys (A1, A3, A5) lead to a significant loss in excursion efficiency. The intact three pulley systems of A2, A3, and A4 were near normal and statistically better than A2 and A4 together for work efficiency. Cutting one of the major pulleys (A2, A4) resulted in significant changes in efficiency, but what was unexpected was to find an 85% loss of both work and excursion efficiency for the loss of A4 but only an excursion difference of 94% for the loss of A2. Our findings demonstrated that in this model, with the influence of the skin removed, A4 absence produced the largest biomechanically measured efficiency changes and that a combination of A2, A3, and A4 was necessary to preserve both work and excursion efficiency. PMID- 8724479 TI - Magnetic resonance imaging scanning in the diagnosis of zone II flexor tendon rupture. AB - This study was undertaken to determine the usefulness of magnetic resonance imaging (MRI) in the diagnosis of flexor tendon rupture in patients who had prior surgery. Magnetic resonance imaging scans were performed on 11 digits (16 tendons) with the clinical diagnosis of flexor tendon rupture. Clinical suspicion correlated with MRI and surgical findings. Clinical examination yielded a 60% accuracy in diagnosis. MRI differentiated rupture from adhesions with a 100% accuracy rate. The MRI scan is a valuable tool in diagnosing tendon ruptures and may help reduce the incidence of unnecessary tendon explorations. PMID- 8724480 TI - Effect of motion and tension on injured flexor tendons in chickens. AB - This study was undertaken to evaluate the individual effects of motion and tension on the healing response of injured flexor profundus tendons in chickens. Partial midsection transverse lacerations of the profundus tendons were produced in 53 chickens. Postoperatively, they were assigned randomly to four management groups: both motion and tension; only motion, no tension; no motion, only tension; no motion and no tension. Biomechanical results at 4 weeks showed that breaking strength significantly increased with both motion and tension, significantly decreased with neither, and was intermediate with only motion or only tension. Histologic evaluation generally showed the greatest cellular activity with both motion and tension, the least with neither, and an intermediate response with one or the other alone. Collagen fiber staining was increased primarily in the tension groups. The finding that both motion and tension enhance the tendon's response to injury encourages the development of active mobilization protocols following tendon repair. PMID- 8724481 TI - Dislocation of the extensor tendons over the metacarpophalangeal joints. AB - Twenty-seven patients who had received treatment for dislocation of the extensor tendons over the metacarpophalangeal joint were reviewed. Sixteen patients had traumatic dislocations, seven had spontaneous dislocations, and four had congenital dislocations. The long finger was most frequently affected. The other fingers were affected almost equally. Displacement of the extensor tendon always occurred in the ulnar direction in the long and ring fingers. The index and little fingers exhibited different patterns of dislocation: two patients had ulnar dislocation of both the common and proprius tendons, and the remaining five patients had divergent dislocation of the two tendons. Nonsurgical treatment was undertaken in six cases. Surgery was performed in 21 cases. No recurrent dislocations were reported in any of the patients. Based on our experience, patients seen within 2 weeks of injury initially should be treated with splinting of the involved metacarpophalangeal joint. Chronic dislocations should be treated with a primary repair of the defect in the sagittal band. When the sagittal band is absent or deficient, the tendon must be stabilized using a loop procedure with a tendon slip. PMID- 8724482 TI - Use of forearm flaps for the severely contracted first web space in children with congenital malformations. AB - Eighteen forearm flaps were used in 12 patients with congenital anomalies following release of severe first web-space contractures. Radial forearm axial pattern flaps were preferred in 10 children, and the dorsal interosseous system was used in 2 patients with incomplete palmar arches and those who had previous surgery. Ten of the flaps were transferred in children younger than 2 years of age. The results were excellent both esthetically and functionally. Superficial skin loss occurred in one patient. These local vascular island flaps are an excellent alternative for coverage in situations where a large tissue deficiency exists after an adequate release. PMID- 8724483 TI - Long-term results of surgical treatment of thumb polydactyly. AB - One hundred thirteen hands exhibiting thumb polydactyly were treated and followed up for an average of 49 months. Of these, 109 hands were treated by resection of a supernumerary hypoplastic thumb. Radial thumbs were resected in 107 hands and ulnar thumbs in 2 hands. Four hands were treated using a modified Bilhaut procedure. According to a modified Tada's evaluation, the results were evaluated as good in 97 hands, fair in 12 hands, and poor in 4 hands. Patients and/or their parents were satisfied with the results in 100 hands and dissatisfied with the results in 13 hands. The factors that influenced the surgical results were analyzed. The incidence of unsatisfactory results was relatively high in Wassel types 3, 5, and 6 and triphalangeal-type thumb polydactyly. It was higher when the ulnar digit was removed than when the radial digit was removed. The results for those patients treated between 1983 and 1991 were better than for those treated between 1976 and 1982. The type of deformity, type of procedure, and skillfulness of the surgeon were factors in the results after surgery. PMID- 8724484 TI - Varicella gangrenosa in the newborn upper extremity: a case report. PMID- 8724485 TI - Collagenase in the treatment of Dupuytren's disease: an in vitro study. AB - The effects of clostridial collagenase on the tensile strength of Dupuytren's cords was studied in vitro to assess its potential efficacy as an agent for clinical enzymatic fasciotomy. Collagenase was injected into Dupuytren's cords from patients undergoing fascioctomy. Following a pilot experiment, in which a 3,600-unit dose of collagenase induced a 93% decrease in tensile modulus as compared with control cords, groups of five cords each were injected with 150, 300, and 600 units. These cords and a control group of five cords were tested by loading to failure in tension. The ultimate stress and strain to failure were recorded by a video capture technique. All specimens were stained for histologic examination with hematoxylin and eosin for collagen typing with sirrius red. Comparison of the ultimate stress values obtained with published values of extensor forces obtainable by the individual fingers of 40 normal hands indicated that a 300-unit dose of collagenase was sufficient for cord rupture within the average maximum force limits of the extensors of the index, long, ring, and small fingers (p < .02). All samples were in the residual disease stage histologically and contained type I collagen by sirrius red staining. These results indicate that collagenase may be effective in enzymatic fasciotomy of residual-stage Dupuytren's disease. PMID- 8724486 TI - In vivo inflammatory response to silicone elastomer particulate debris. AB - Silicone elastomer particles (Silastic silicone elastomer, Dow Corning, Midland, MI), polymethylmethacrylate particles, and monosodium urate particles smaller than 10 microns were injected into a rat subcutaneous air pouch lined with synovial membrane-like cells. Inflammatory exudate from the air pouch was retrieved at 6 hours, 24 hours, 48 hours, and 72 hours after injection. White blood cell count, tumor necrosis factor, and prostaglandin E2 were measured in the exudate. White blood cell and tumor necrosis factor levels in the exudate were the highest for the silicone group at 6 and 24 hours. Prostaglandin E2 was also significantly higher in the silicone group at 24 hours. We conclude that the acute inflammation is particle-type specific and that Silicone elastomer particles are acutely inflammatory. PMID- 8724487 TI - Florid reactive periostitis of the digit with cortical erosion: a case report and review of the literature. PMID- 8724488 TI - Subungual melanoma of the hand. AB - Subungual melanoma is a rare but well-recognized tumor of the hand. Its management is ill defined and the factors influencing prognosis have not been well described. The clinicopathologic features of a series of 38 patients with subungual melanoma of the hand are reported. The median thickness was 3 mm, and only seven patients presented with pathologic stage I disease (American Joint Committee on Cancer [AJCC] system). Ulceration and lack of pigmentation were the only significant univariate prognostic indicators. There was no significant difference in local recurrence rates among patients whose amputation was carried out proximal or distal to the interphalangeal joint of the thumb or the middle of the middle phalanx in the other fingers. Management of the regional lymph node field based on the use of selective lymphadenectomy is described. PMID- 8724489 TI - Subungual lipoma, squamous carcinoma of the nail bed, and secondary chronic infection. PMID- 8724490 TI - Actinomycetoma of the phalanx in an immunocompromised patient: a case report. PMID- 8724491 TI - Electrical studies as a prognostic factor in the surgical treatment of carpal tunnel syndrome. PMID- 8724492 TI - Electric studies as a prognostic factor in the surgical treatment of carpal tunnel syndrome surgery. PMID- 8724493 TI - Electric studies as a prognostic factor in the surgical treatment of carpal tunnel surgery. PMID- 8724494 TI - Electrical studies as a prognostic factor in the surgical treatment of carpal tunnel syndrome. PMID- 8724495 TI - Electrical studies as a prognostic factor in the surgical treatment of carpal tunnel syndrome. PMID- 8724496 TI - Electrical studies as a prognostic factor in the surgical treatment of carpal tunnel syndrome. PMID- 8724497 TI - Electrical studies as a prognostic factor in the surgical treatment of carpal tunnel syndrome. PMID- 8724498 TI - Outcome following conservative management of thoracic outlet syndrome. PMID- 8724499 TI - The second annular pulley: a histologic examination. PMID- 8724500 TI - Steroid versus placebo injection for trigger finger. PMID- 8724501 TI - Pinpointing the moment of allograft rejection. PMID- 8724502 TI - Transplantation of second and third toes to the hand. PMID- 8724503 TI - Acyclovir should not be approved for marketing without prescription. PMID- 8724504 TI - Over-the-counter acyclovir. An idea whose time has come. PMID- 8724505 TI - Detection of Chlamydia trachomatis by a ligase chain reaction amplification method. AB - BACKGROUND AND OBJECTIVES: The ligase chain reaction is an in vitro DNA amplification technique that exponentially amplifies selected DNA sequences. GOAL: To evaluate a ligase chain reaction assay for the detection of Chlamydia trachomatis cryptic plasmid DNA (LCx Chlamydia) in patients routinely attending a sexually transmitted disease center in Italy. STUDY DESIGN: Urethral or cervical swabs were obtained from 501 consecutive patients (334 men and 167 women). The samples were assayed in parallel with LCx Chlamydia and conventional tissue culture; discordant results were further assayed by direct immunofluorescence and a ligase chain reaction with alternate primers. RESULTS: After resolution of discordant results, the LCx method showed a sensitivity, specificity, positive predictive value, and negative predictive value of 100%, 99.3%, 96.7%, and 100% in men; 100%, 100%, 100%, and 100% in women; and 100%, 99.5%, 97.1%, and 100% overall, respectively. By comparison, the sensitivity of tissue culture was 81.4% in men, 50% in women, and 77.6% overall. CONCLUSIONS: The automated LCx method is sensitive, fast, and accurate and represents a useful diagnostic tool for C. trachomatis infection, even in low and medium prevalence populations. PMID- 8724506 TI - Gonococcal endocarditis. PMID- 8724507 TI - Resolution of serum and cerebrospinal fluid abnormalities after treatment of neurosyphilis. Influence of concomitant human immunodeficiency virus infection. AB - BACKGROUND AND OBJECTIVES: Little is known about resolution of serum and cerebrospinal fluid abnormalities after neurosyphilis treatment, especially in patients infected with human immunodeficiency virus (HIV). GOAL: To examine the time course of resolution of these abnormalities. STUDY DESIGN: Case series of 22 patients with neurosyphilis (13 infected with HIV) with reactive cerebrospinal fluid Venereal Disease Research Laboratory test who underwent at least one lumbar puncture after treatment. RESULTS: Resolution of all serum and cerebrospinal fluid measures was slower in patients infected with HIV. Serum and cerebrospinal fluid abnormalities resolved in most patients not infected with HIV by 30 weeks, and all met Centers for Disease Control and Prevention criteria for cure. One patient infected with HIV failed therapy by Centers for Disease Control and Prevention criteria, and three others had persistent pleocytosis. CONCLUSIONS: HIV-infected patients with neurosyphilis have slower resolution of serum and cerebrospinal fluid abnormalities after therapy. This observation may suggest impaired clearance of Treponema pallidum from the central nervous system. PMID- 8724508 TI - Usefulness of human immunodeficiency virus post-test counseling by telephone for low-risk clients of an urban sexually transmitted diseases clinic. AB - BACKGROUND: Failure of clients to return for post-test counseling for human immunodeficiency virus (HIV) has been noted consistently as a problem with HIV counseling and testing in sexually transmitted disease clinics. GOALS: To assess trends in the rates of HIV post-test counseling in an urban sexually transmitted disease clinic. To determine the usefulness of providing post-test counseling by telephone to clients at low risk for HIV as a means of increasing post-test counseling rates and efficiency of clinic operations. STUDY DESIGN AND METHODS: Human immunodeficiency virus post-test counseling rates were evaluated among clients of a sexually transmitted disease clinic from January 1990 through May 1994. Low-risk clients (n = 1,304) from July 1994 through September 1994 were given the option of obtaining negative HIV test results by telephone. Rates of post-test counseling within 45 days were compared with historical controls from March 1994 through May 1994, during which time a return visit was required. RESULTS: The rate of post-test counseling increased significantly from 1990 to 1992 but remained stable thereafter. After the introduction of post-test counseling by telephone, 704 of 1,304 low-risk clients (54.0%) obtained post-test counseling compared with 476 of 1,187 (40.1%) clients during the control period (odds ratio = 1.75; 95% confidence interval = 1.50-2.06). Post-test counseling rates increased especially among clients younger than 20 years of age (30.6% versus 52.7%) and among those with multiple new sex partners in the last month (31.6% versus 56.1%). CONCLUSIONS: Telephone post-test counseling is an effective method of increasing access for low-risk clients attending sexually transmitted diseases clinics and may be especially useful for those who are otherwise unlikely to obtain post-test counseling. PMID- 8724509 TI - Epidemiologic risk factors for incident sexually transmitted diseases in young Thai men. AB - BACKGROUND AND OBJECTIVES: Prospective studies of incidental sexually transmitted diseases in developing countries rarely have been reported. As the human immunodeficiency virus epidemic widens, knowledge of the epidemiology of sexually transmitted diseases as co-factors for human immunodeficiency virus infection is viewed as vitally important. GOAL: To determine epidemiologic risk factors for incidental sexually transmitted diseases in young men in northern Thailand. STUDY DESIGN: A cohort of 2,417 men selected by lottery for military conscription was followed an average of 22 months, with epidemiologic risk factors and sexually transmitted disease histories assessed semiannually. Sexually transmitted diseases were determined by physician or symptom histories obtained by interview. RESULTS: Sexually transmitted disease incidence was 17.04 per 100 person-years. Gonorrhea and chancroid were commonly reported, whereas incident syphilis was rare. Sexually transmitted disease incidence declined significantly in the 2-year period. Most sexually transmitted diseases were acquired through commercial sex patronage. However, among men who did not report visiting a brothel, there was a significant increase in sexually transmitted diseases among men reporting sex with a girlfriend. Other risk factors included inconsistent condom use, drug use, and low educational level. Frequent alcohol use was associated with incident sexually transmitted diseases in bivariate analysis, but drinking at the time of last brothel visit was not associated with increased sexually transmitted diseases. CONCLUSIONS: Sexually transmitted disease control in Thailand is based on an integrated sexually transmitted disease clinic system. Many commercial sex workers are examined and treated routinely, but too infrequently to reduce sexually transmitted disease transmission between them and their male clients. Condom-use levels in commercial sex settings still may be too low to reduce sexually transmitted disease transmission. More consistent and effective sexually transmitted disease treatment of commercial sex workers and their clients and education of men on the effective and consistent use of condoms is required to reduce the risks of sexually transmitted disease transmission in this population. PMID- 8724511 TI - Gonococcal susceptibility to antimicrobials in Baltimore, 1988-1994. What was the impact of ciprofloxacin as first-line therapy for gonorrhea? AB - BACKGROUND: Single-dose, oral quinolones have been a recommended treatment option for gonorrhea since 1989 and have since been used as first-line therapy for gonorrhea in Baltimore. METHODS: The impact of this strategy on antimicobial susceptibility patterns in Neisseria gonorrhoeae was assessed utilizing data collected as part of the National Gonococcal Isolate Surveillance system. This system evaluates a systematically collected sample. Minimum inhibitory concentrations to penicillin, tetracycline, ceftriaxone, and ciprofloxacin were determined by agar dilution. RESULTS: Between January 1988 and September 1994, 1,846 gonococcal isolates were evaluated. The proportion of isolates with plasmid mediated resistance (penicillinase-producing Neisseria gonorrhoeae or tetracycline-resistant Neisseria gonorrhoeae) increased from 22% in 1988 to 46% in 1992 and then dropped to 20% in 1994. In contrast, the prevalence of chromosomally mediated resistant isolates ranged between 3% and 10%. Between 1988 and 1994, the geometric mean penicillin and tetracycline minimum inhibitory concentrations decreased slightly (penicillin: from 0.509 microgram/ml to 0.369 microgram/ml; tetracycline: from 1.01 micrograms/ml to 0.767 microgram/ml). The mean ceftriaxone MIC increased from 0.005 microgram/ml in 1988 to 0.021 microgram/ml in 1992, and then abruptly decreased. Ciprofloxacin minimum inhibitory concentrations did not change substantially during the study period. Concurrent studies performed in this community suggest that quinolones were infrequently used for infections other than sexually transmitted ones during this time period. CONCLUSIONS: Single-dose quinolone therapy does not appear to foster development of resistant gonococcal isolates. However, resistance may develop as a result of complex ecological interactions with the community, underscoring the need for continued surveillance. PMID- 8724510 TI - Use of spermicide and impact of prophylactic condom use among sex workers in Santa Fe de Bogota, Colombia. AB - BACKGROUND AND OBJECTIVES: Condom use is one of the most important preventive measures sex workers can take to reduce the risk of becoming infected with a sexually transmitted disease. However, a client may refuse to use a condom when requested. Some sexually transmitted disease prevention programs are recommending that sex workers use spermicide as an alternative prophylaxis when a condom is refused, yet little is known about the effect of this recommendation on prophylactic condom use. GOAL: To determine if using spermicide, either in conjunction with condoms or as a backup, influenced overall condom use among a group of sex workers at high risk of sexually transmitted diseases in Santa Fe de Bogota, Colombia. STUDY DESIGN: Participants were assigned randomly to one of three condom use groups: use of condoms only (Condoms Only), use of condoms and spermicides concurrently (Condom and Spermicide), or use of spermicide when condoms were refused (Spermicide as a Backup). A total of 199 sex workers entered the study and were asked to return for follow-up every 2 weeks for a period of 12 weeks. RESULTS: Women assigned to the Spermicide as a Backup group used a condom for an average of 78.1% of their reported acts of intercourse, compared with an average of 94.5% in the Condom Only and 92.3% in the Condom and Spermicide groups. However, women in the Spermicide as a Backup group used a condom or spermicide for an average of 96.9% of their acts of intercourse. Condoms were used for every intercourse act by less than 5% of the women in the Spermicide as a Backup groups, compared with 50.7% in the Condom Only group and 41.2% in the Condom and Spermicide group (P 0.001). When condoms were not used, client refusal was the primary reason reported. The incidence of sexually transmitted diseases and other urogenital inflammations in all groups was lower than expected. CONCLUSIONS: Among Colombian sex workers, condom use declined substantially when women were instructed to use spermicides if they were unable to persuade their partner to use a condom. However, these same women usually used the study spermicide as an alternate prophylaxis. PMID- 8724512 TI - Acquired immune deficiency syndrome. Knowledge, experiences, and attitudes of hospital-based registered nurses. AB - BACKGROUND AND OBJECTIVES: Because of the rapidly increasing number of people affected by acquired immune deficiency syndrome, an increasing number of health care providers are involved. The knowledge, experiences, and attitudes of hospital-based registered nurses can affect the quality and quantity of care provided people with acquired immune deficiency syndrome. GOALS: To determine the knowledge, experiences, and attitudes about acquired immune deficiency syndrome of registered nurses employed in hospitals in Oklahoma and delineate changes in that knowledge and attitudes over time. This should provide a measure of the preparedness of hospital-based registered nurses in Oklahoma to facilitate initial and continued care of people with acquired immune deficiency syndrome. STUDY DESIGN: This study surveyed hospital-based registered nurses in Oklahoma in 1986 and again in 1994. In 1986, 75 (51%) of the hospitals and 446 (68% response rate) of the registered nurses participated in the study. In 1994, 85 (53%) of the hospitals and 564 (74% response rate) of the registered nurses participated in the study. The study sample demographics closely matched the state and national populations of hospital-based registered nurses. RESULTS: The average scores on general and clinical knowledge tests increased significantly from 1986 to 1994. In general, infection control registered nurses had statistically significantly greater knowledge than did registered nurses assigned to other areas of work. However, deficiencies were evident in specific areas of knowledge. The attitudes of registered nurses toward people with acquired immune deficiency syndrome have become more positive since 1986. Some confusion and unjustifiable fear may exist among some registered nurses regarding certain aspects of acquired immune deficiency syndrome. CONCLUSIONS: The hospital-based registered nurses in Oklahoma have increased their preparedness to facilitate initial and continued care of people with acquired immune deficiency syndrome. Because of the representative sample of this study, that conclusion probably is applicable to the nation. PMID- 8724513 TI - Declining prevalence of chlamydial infection among adolescent girls. AB - BACKGROUND AND OBJECTIVES: Chlamydial control programs that identify and treat infected persons have been used as a means of reducing prevalence and morbidity. The overall objective of the current study was to examine the prevalence of chlamydial infection during the course of such a program. GOALS: To determine whether a reduction in isolation rates was evident among adolescent girls during the 8.75 years that the control program has been in place. STUDY DESIGN: The study population consisted of all 4,329 sexually active girls between the ages of 13 and 19 attending the four adolescent health clinics in Indianapolis, Indiana, during the period beginning October 1, 1985, and ending June 30, 1994. All girls were cultured for Chlamydia trachomatis, and behavioral data were collected for those attending the clinics before 1989. The trend in quarterly isolation rates was examined using linear regression analysis. RESULTS: Results showed that there was a significant decrease (P = 0.0001), from 25.9% to 9.7%, in the first-visit chlamydial isolation rate over the study period. Behavioral data showed decreases in the frequency of sexual intercourse and in lifetime years of sexual activity, as well as an increase in condom use. CONCLUSIONS: The quarterly isolation rates showed that there has been a 63% decline in chlamydial infection among adolescent girls attending the clinics for the first time. PMID- 8724514 TI - Trends in the acquisition of sexually transmitted diseases among HIV-positive patients at STD clinics, Miami 1988-1992. AB - BACKGROUND AND OBJECTIVES: To assess trends in the acquisition of new sexually transmitted diseases (STDs) among patients who test positive for human immunodeficiency virus (HIV) at STD clinics. STUDY DESIGN: Cohorts of HIV positive and HIV-negative persons were compared using computerized records from Miami STD clinics for 1988-1992. Persons were assigned to cohorts according to their first positive or first negative HIV test results. New STDs were defined if persons had new diagnoses of gonorrhea, primary or secondary syphilis, chancroid, or lymphogranuloma venereum; were undergoing treatment as contacts for syphilis or gonorrhea; or were undergoing epidemiologic treatment for syphilis or gonorrhea. RESULTS: Of the 103,549 persons who visited the clinics, 53,467 were tested for HIV, and 5,615 had results that were positive. The percentages returning with new STDs were similar for the HIV-positive and HIV-negative cohorts, and both decreased over time. For the 1988 cohorts, 26% of those testing positive and 30% of those testing negative for HIV returned with at least one STD within 5 years. Returns with STD within 1 year decreased from 16% in 1988 to 3% in 1992. CONCLUSIONS: Returns decreased dramatically among HIV-positive cohorts; however, there were similar decreases of new STDs among HIV-negative cohorts, so the decrease may have been caused by the decreasing prevalence of bacterial STD in the community rather than by behavioral changes among HIV-positive persons. PMID- 8724515 TI - Comparison of development of serum antibodies to HPV16 and HPV33 and acquisition of cervical HPV DNA among sexually experienced and virginal young girls. A longitudinal cohort study. AB - OBJECTIVES: To study the importance of sexual activity and early coitus debut on the risk for acquiring infection with human papillomavirus (HPV) type 16 or 33. STUDY DESIGN: Ninety-eight healthy adolescent girls were followed up with consecutive interviews and donations of serum and cervical brush samples during 2 years. RESULTS: Fourteen percent of sexually experienced girls had serum immunoglobulin G to HPV16 and/or HPV33 capsids, and 14% also had cervical HPV16 or HPV33 DNA. Seropositivity for HPV correlated with detection of cervical HPV DNA. None of the 36 girls without coital experience was seropositive or harbored cervical HPV DNA. Seropositivity for HPV was correlated strongly with the number of sexual partners: Odds ratio for > 1 sexual partner was 16.3 (P < 0.001), and for early coitus debut (younger than 17 years of age), it was 14.3 (P < 0.002). CONCLUSIONS: Both HPV serology and HPV DNA testing indicated that the number of sexual partners and earliness of coitus debut determined the risk for acquiring HPV infection and that nonsexually transmitted infections are rare or nonexistent among adolescent girls. PMID- 8724516 TI - Risk factors for pelvic inflammatory disease. A case control study. AB - BACKGROUND AND OBJECTIVES: Pelvic inflammatory disease (PID) is associated with major medical and economic consequences for women of reproductive age. Identification of the risk factors associated with PID is crucial to efforts for prevention of these consequences. GOAL: To identify risk factors and markers for symptomatic PID. STUDY DESIGN: A case-control study of 234 women with PID treated at San Francisco General Hospital between October 1986 and August 1989 and 122 controls attending the Women's Clinic at the same institution. The two groups were comparable in sociodemographic, reproductive, sexual, and medical history. RESULTS: Risk factors and markers identified by univariate analysis were < 12 years education, gravidity > 0, parity > 0, spontaneous abortion > 0, lack of a birth control method, > 1 male sexual partner in the previous 30 days, younger than 18 years at age of first sex, history of gonorrhea, sex during the previous menses, douching, exposure to nongonococcal urethritis in the previous 30 days, and history of crack cocaine use. With multivariate analysis to control for confounders the risks still identified were parity > 0, (odds ratio [OR] 4.44; 95% confidence interval [CI] 2.34 to 8.42), > 1 sexual partner in the previous 30 days (OR 11.08; 95% CI 4.31 to 28.5), sex during the previous menses (OR 5.22; 95% CI 1.88 to 14.48), and a lack of contraception (OR 7.6; 95% CI 4.10 to 14.09). CONCLUSIONS: Findings indicated that certain reproductive behaviors could be targeted for public health attention and risk reduction interventions to reduce the incidence of PID. These include limiting numbers of sexual partners and encouraging the use of barrier methods of contraception for sexually transmitted disease prevention. Another finding was that it is probably best to avoid sexual intercourse during the menses. The question of douching as a risk factor for PID could not be answered by this study. PMID- 8724517 TI - Sexual mixing patterns of patients attending sexually transmitted diseases clinics. AB - BACKGROUND: Theorectical studies have highlighted the importance of patterns of choice of sex partner in the transmission and persistence of sexually transmitted diseases (STDs). GOAL: To describe reported patterns of sexual mixing according to numbers of sex partners in STD clinics. STUDY DESIGN: Patients attending public health clinics in Seattle, Washington were interviewed about their own and their partners' behaviors. RESULTS: Throughout, patterns of sexual mixing were weakly assortative. Across activity groups, many respondents believed their partners had no other sexual contacts. Those with three or more partners frequently perceived their partners to have three or more partners as well. CONCLUSIONS: Assortatively mixing persons of high sexual activity makes the persistence of STDs within a population likely (i.e., they act as a "core group"). Additionally, because mixing is not highly assortative (like with like), a steady trickle of infection from members of the core group will pass to other segments of the population. PMID- 8724519 TI - A non-invasive, three-dimensional, diagnostic laser imaging system for accurate wound analysis. AB - A PC-based laser imaging system has been developed that can accurately image and analyse three-dimensional (3D) surfaces in healing wound tissue. The device consists of a laser diode and sensor mounted to a motorized, bi-directional, X-Y table. The laser measures the distance to the wound surface as the X-Y table moves over the wound. The laser beam diameter is 1 x 2 mm and can measure distances as small as 8 microns. The distance information from the laser is used to produce the 3D image and to calculate the volume of the wound. The accuracy of this device is demonstrated by measuring rubber model wounds of 0.5, 1 and 2 ml. The per cent error for the 0.5 and 1 ml model wounds was less than 1 per cent and approximately 4 per cent for the 2 ml model. Studies were done in which full thickness wounds were made on rats and monitored over a two week period. The rats were scanned five times during this period and the wound area and volumes calculated. The laser measurements are compared to measurements using saline and wound thickness multiplied by wound area. PMID- 8724518 TI - Surgical wound healing monitored repeatedly in vivo using electrical resistance of the epidermis. AB - We describe a new transcutaneous electrical resistance method for measuring surgical wound healing which permits repeated measurements in vivo throughout the healing period. A saline-filled silver electrode was used to monitor the increasing ohmic resistance which accompanied the healing. Two validation studies were carried out. In the first study, histology on 16 rats showed that an intact stratum corneum was responsible for the high electrical resistance of skin, since lesions of the corneum markedly lowered the resistance. In the second study, electrical resistance readings of incisional wounds were taken repeatedly over 20 days in 13 rats: regression analysis showed that the healing slope for each incisional wound had a correlation coefficient r-value greater than 0.85. Moreover, all slopes were positive (p < 0.001) demonstrating that resistance increases as wound healing progresses and hence is a valid measure of healing. Test-retest resistance readings showed high reliability when measurements were repeated on the same wound (Pearson r = 0.94, n = 110). Hence we have shown that this new method is both valid and reliable. The device only monitors epidermal repair and gives no indication of wound strength. A modified device recently developed for humans has been used successfully on post-surgical wounds in numerous patients. PMID- 8724520 TI - The origin of skin-stretch-caused motion artifacts under electrodes. AB - The outside of the skin of the forearm is typically 15 mV more negative than the inside. Stretching the skin causes a reduction in the magnitude of this skin potential V, which we observe as a motion artifact delta V. We seek to determine the origin of this motion artifact by successively stripping 12 layers of the skin using Scotch Tape. Between each stripping we measure artifact delta V, 13 Hz impedance Z, and change in impedance delta Z. On the interior surface of the forearm, Z decreases with number of strippings. delta Z can be first either positive or negative, then is always negative and decreases linearly with Z. delta V first remains constant and then decreases with Z and delta Z. delta V and delta Z increase with stretch force following a logarithmic relationship. delta Z has a rectangular shape waveform, whereas the rising edge of delta V shows a fast followed by a slow component and its falling edge decays exponentially with a large time constant. We have expanded the model of Thakor and Webster to best fit the waveform of delta V and delta Z caused by stretch. PMID- 8724521 TI - Automated determination of the left ventricular long axis in cardiac positron tomography. AB - Interpretation of emission tomographic images of the heart is typically performed using short-axis sections which are oriented perpendicular to the long axis of the left ventricle. A completely automated method is presented to find the orientation and length of the long axis in positron emission tomographic studies of the heart. The correlation coefficient is maximized between the measured transaxial images and an ellipsoid model of the left ventricular myocardium. The major axis of this fitted ellipsoid corresponds to the long axis of the left ventricle. The orientation and position of the long axis (with respect to the measured transaxial images) define two rotation angles and a 3D coordinate origin which are used to re-orient the transaxial images into a series of standard short axis sections. The accuracy and precision of this technique are validated on phantom data as well as on patients with documented myocardial infarction. The transaxial and vertical long-axis rotation angles are determined with standard deviations of 3.3 degrees and 1.5 degrees, respectively. The 3D coordinate origin (centre) of the ellipsoid is accurate to within 1.5 mm on average. The estimated length of the left ventricle is accurate to within 3 mm. All parameters are insensitive to statistical noise found in typical 18F-fluorodeoxyglucose (FDG) patient studies. The technique produces accurate estimates even in the presence of moderate uptake defects in the lateral wall and septum. Apical defects in the uptake of FDG do not increase the variance of the length estimate. These results demonstrate that an ellipsoid model can be fitted accurately to the myocardium of the left ventricle. Standard short-axis sections are produced with no inter operator or intra-operator variability because the technique is fully automatic. PMID- 8724522 TI - Simultaneous electrical impedance tomography and videofluoroscopy in the assessment of swallowing. AB - Simultaneous videofluoroscopy and electrical impedance tomography (EIT) were performed in 13 patients. Subjects swallowed a low-conductivity and a higher conductivity bolus of between 5 and 20 ml. EIT images from the two swallows were then subtracted to cancel out movement related changes. The peak in the conductivity change was most closely associated in time with a point at which the pharynx is filled with bolus: this suggests that the subtraction process does produce data and images which are related to the bolus rather than to movement of involved structures. The full width at 50% and 75% of the maximum of the time conductivity plot was correlated with the time taken for the bolus to be cleared from the mouth (r = 0.63 and 0.68 respectively, p < 0.05 for both). Despite the sampling frequency of only 9 frames s-1, compared to 50 frames s-1 for videofluoroscopy, EIT recordings from the neck are able to detect the presence of a conductive bolus in the pharynx and for the first time the relationship between parameters of an EIT time-conductivity plot and a measure of bolus transit using an accepted technique has been established. PMID- 8724523 TI - New perspectives on the approach to chronic lymphocytic leukemia. AB - Investigation of the biological actions of loxoribine in chronic lymphocytic leukemia (CLL) was undertaken because of the pervasive immunostimulatory effects of the nucleoside on normal B cells. In vitro studies with cells from a spectrum of CLL patients demonstrate that loxoribine induces B-CLL cells to enter and traverse the cell cycle. This is reflected by marked increases in DNA synthesis, by standard morphological criteria, and by flow cytometric evaluation of cell cycle status and of cell surface activation markers. Cells from about 75% of patients studied evince this response. Analysis of a variety of biological parameters indicate that only the ratio of T cells (CD4+ or CD8+) to B-CLL cells correlates with induction and degree of proliferative response. Co-stimulation with loxoribine and IL-2 results in modest proliferative synergy, presumably due to upregulation of IL-2R alpha expression on B-CLL cells by loxoribine. Prolonged exposure of B-CLL cells to stimulatory concentrations of loxoribine frequently culminates in progression of the responsive cells to apoptosis. The capacity of loxoribine to transiently approximate the reversible transformation of a low grade B cell malignancy to one of a higher grade presents the opportunity for evaluation of cycle-active drugs under these conditions. Recent studies indicate that pre-treatment of B-CLL cells with loxoribine results in synergistic killing of leukemic cells with cycle-active drugs. The ability to induce B-CLL cells into cell cycle entry and/or into either activation-induced apoptosis or into phases of the cell cycle sensitive to cytotoxic therapy opens up new perspectives for the development of potentially curative strategies for this chronic leukemia. PMID- 8724524 TI - p16ink4a gene and hematological malignancies. AB - Progression of eukaryotic cells through major cell cycle transitions is mediated by sequential assembly and activation of regulators, the cyclin-dependent CDKkinases (CDKs). Recent studies have identified different CDK inhibitory genes (CDKis), and two of them, p16ink4a/MTS1/CDKN2 and p15ink4b/MTS2 are both mapped to chromosome 9p21 and inhibit cyclin D-CDK4 and -CDK6 complexes. A feedback regulatory loop involving pRb, p16ink4a, and CDKs seems to regulate G1/S phases transition. p16ink4a and p15ink4b are deleted in high frequency in human cell lines and in some fresh solid tumors. Point mutations of p16ink4a have also been sequenced, especially in familial melanomas and digestive cancers but preferential mechanism of p16ink4a/p15ink4b inactivation seems to be biallelic deletion. In hematological malignancies, homozygous deletions of p16ink4a and p15ink4b occur frequently in acute lymphoblastic leukemia (ALL) (14-40%), lymphoid type blast crisis of chronic myeloid leukemia (CML), and adult T cell leukemia (ATL), but p16ink4a deletions are more frequent than p15ink4b deletions, and hemizygous deletions of either p16ink4a and p15ink4b are rare. In ALL an association of homozygous deletions of p16ink4a and p15ink4b, and T-lineage, 9p abnormalities, and prognostic factors was found in some but not all reports. This review presents recent data on p16ink4a and p15ink4b functions and analyses their implications in hematological malignancies. PMID- 8724525 TI - Hematopoietic stem cell transplants for multiple myeloma. AB - Standard chemotherapy with melphalan-prednisone or a combination of alkylating agents has not extended the overall survival of patients with multiple myeloma during the last 30 years and strictly defined complete remissions (CR) are exceedingly rare. The early mortality with conventional therapy varies between 2 and 10 percent. A substantial increase in the dose of melphalan (100-140 mg/m2) has resulted in a 30-45% CR rate in newly diagnosed patients and an overall survival advantage of approximately 1 year. However, treatment related morbidity and mortality, due to prolonged cytopenia was unacceptably high. Based on these findings the dose intensity was further increased by either escalating melphalan to 200 mg/m2 or by adding total body irradiation, while at the same time providing stem cell support to shorten the duration of cytopenia. Autologous transplants, especially with peripheral blood stem cells and hematopoietic growth factors, can now be performed safely up to the age of 70 with a low transplant related mortality (2-10%). A CR is attained in approximately 50% of previously untreated patients and 10-20% of refractory cases. Overall survival of newly diagnosed and refractory patients treated with autotransplants appears superior to that of patients receiving conventional chemotherapy. Therefore, autotransplantation should be considered as a treatment option in all patients with multiple myeloma at least up to the age of 65. Despite these encouraging findings, most myeloma patients ultimately relapse and the survival curves do not suggest that autotransplantation as currently performed is a curative approach in a substantial proportion of patients. Further improvement with autotransplants should be achieved by providing tumor-free grafts and by introducing post transplantation manipulations, aimed at eradicating minimal residual disease. PMID- 8724526 TI - CD34+Thy-1+Lin- stem cells from mobilized peripheral blood. AB - Over the last ten years there has been increasing use of mobilized peripheral blood (MPB) progenitor cells as grafts for autologous transplantation. Among the cells comprising these MPB autografts is a subpopulation of CD34+Thy-1+Lineage (Lin)- cells, which is enriched for hematopoietic stem cell (HSC) activity. The percentage of CD34+ cells which express Thy-1 is higher in some samples of MPB than in bone marrow (BM). Using myeloid and erythroid cell depletion prior to high speed cell sorting, it is possible to purify sufficient numbers of CD34+Thy 1+Lin-HSCs from a MPB leukapheresis sample for use as an autograft. CD34+Thy 1+Lin-cells will potentially provide a tumor-depleted autograft for cancer patients. This HSC population is also being studied as a potential target for gene transfer for the treatment of patients with HIV, cancer and a variety of genetic disorders. PMID- 8724527 TI - Proliferation and differentiation of myelodysplastic CD34+ cells. AB - Myelodysplastic syndromes (MDS) are a heterogeneous group of disorders of hematopoiesis involving hyperproliferative and ineffective hematopoiesis associated with morphologic evidence of marrow cell dysplasia resulting in refractory cytopenia(s), and an increased risk of transformation into acute myeloblastic leukemia (AML). The administration of colony-stimulating factor(s) (CSFs) to patients with MDS increased blood neutrophil concentrations, in most patients, and was also expected to be beneficial and to prevent infections. However, the progression to AML during the treatment with CSFs was suspected in some patients. Therefore, extensive in vitro studies were expected to lead to the establishment of criteria for selection of patients who are likely to benefit from CSF's as well as to establish the overall value of the different types of CSFs therapy. For this purpose, in vitro colony assays provide an excellent tool for investigating the biologic characteristics of MDS progenitor cells. However, conditions of the culture must be such that each progenitor can express its full potential for proliferation and differentiation. Because of the above, MDS progenitor cells cannot be used because they carry an impairment in proliferation and differentiation. To address this problem, one needs to know how many cells are being handled and the maximum numbers of colonies and clusters expected. CD34, a stem cell phenotype, is at present one of the best markers of progenitor cells, and can be used for purposes of purification. Using a defined number of CD34+ cells, it was feasible to make direct investigations on MDS progenitor cells. In this review the properties of MDS progenitor cells are described, in association with proliferation and differentiation, with special emphasis on the phenotypic subpopulations of MDS CD34+ cells. PMID- 8724528 TI - Hepatitis C virus and non-Hodgkin's lymphomas. AB - The etiology of non-Hodgkin's lymphomas (NHL) remains a controversial matter, but, in the last few years, considerable evidence suggests that aberrations of the immune system and viruses may act as etiologic agents, in at least some cases of NHL. In fact, patients with primary immuno-deficiencies, or those suffering from diseases characterized by autoimmune dysfunction, show an increased risk for the development of NHL. Several viruses have been identified as possible etiologic agents for NHL; one of the best studied is the Epstein-Barr virus, which was detected in cultures of tumor cells from patients with Burkitt's lymphoma. The pathogenetic potential of this virus is illustrated by its association with an increasing number of malignant diseases. In addition, the human T-cell lymphotropic virus family (HTLV), was also recognized as possible etiologic agents for several lymphomas, such as cutaneous T-cell lymphoma and T cell leukemia-lymphoma syndrome (HTLV-I), and T-cell hairy cell leukemia (HTLV II). Recently, the presence of hepatitis C virus infection has also been recognized in several hematological malignancies such as mixed cryoglobulinemia, low-grade malignant lymphomas and Waldenstrom's disease. The possible etiopathogenetic role of this virus in non-Hodgkin's lymphomas is discussed on the basis of molecular, clinical, and epidemiological considerations. PMID- 8724529 TI - Toxicity profile of the investigational new biotherapeutic agent, B43 (anti-CD19) pokeweed antiviral protein immunotoxin. AB - The investigational biotherapeutic agent, B43(anti-CD19)-pokeweed antiviral protein (PAP) immunotoxin, has shown substantial anti-leukemic activity in SCID mouse models of human B-lineage leukemia and lymphoma. In this report, we describe the results of a comprehensive preclinical toxicity study which determined the toxicity profile of B43-PAP in BALB/c mice. Administration of unconjugated B43 monoclonal antibody was not associated with any toxicity, whereas B43-PAP caused dose-limiting and cardiac and renal toxicities which were fatal. In addition, B43-PAP also caused multifocal skeletal myofiber necrosis, which was associated with abnormal gait and lethargy. Notably, parenteral administrations of methylprednisolone, pentoxyphylline, or dopamine were able to markedly reduce B43-PAP related toxicity. This study provides a basis for further evaluation of the toxicity of B43-PAP in monkeys and humans. PMID- 8724530 TI - Amsacrine and continuous-infusion high-dose cytosine arabinoside as induction therapy for patients with newly-diagnosed acute myelogenous leukemia. AB - The overall cure rate of adults with newly-diagnosed acute myelogenous leukemia (AML) treated with continuous infusion high-dose cytarabine (CIHDAC) is comparable to that with standard-dose ara-C plus anthracycline or amsacrine (AMSA). We tested whether the addition of AMSA to CIH-DAC improves the outcome of adults with untreated AML. 75 patients with untreated AML were treated with AMSA (75 mg/m2/day x 4) plus CIHDAC (1.5 g/m2/day x 4) for induction and, if in complete remission (CR), early and late intensification. Results were compared to those in 129 patients treated on a previous study with CIHDAC alone. The principal comparison in both groups was between those 117 patients (AMSA/CIHDAC n = 52, CIHDAC n = 65) who met the initial eligibility criteria for the AMSA/CIHDAC study (risk of early mortality < or = 1) and who were treated at a time when relatively few eligible patients were excluded (19% in the AMSA/CIHDAC group, 34% in the CIHDAC group). There was no difference between regimens in CR rate, remission duration, or survival in this cohort. When attention was turned to all 204 patients, outcome was superior with AMSA/CIHDAC very largely as a result of outcome in patients with APL. Aside from these patients, addition of amsacrine to CIHDAC did not appear to be productive. PMID- 8724531 TI - Apoptosis in acute myeloblastic leukemia: follow-up study on trephine biopsies of the bone marrow. AB - A clinicopathological study on 87 adult patients presenting with "de novo" acute myeloblastic leukemia (AML) was performed to assess the rate of apoptosis before and during chemotherapy and its predictive impact on clinical course. Evaluation included trephine biopsies of the bone marrow and the situ end-labeling technic (ISEL) for the identification of programmed cell death in large and intact hemopoietic tissue areas. In comparison with a control group of 21 patients without any hematological disorder, morphometric analysis revealed no significantly different numbers of apoptotic cells in AML at the onset of disease and following sequential examinations at intervals ranging between 10 to 19 months. Moreover, the incidence of programmed cell death was not associated with the subgroups of the FAB classification and statistics failed to show a relationship with survival or remission status. In conclusion, these findings are in keeping with the assumption that apoptosis occurs with the same frequency in recovering normal hemopoiesis in complete or partial remission, in manifest AML and relapse. In the latter conditions, enhancement of proliferation is not associated with an increase in the apoptotic index. PMID- 8724532 TI - Interleukin-6 inhibits apoptosis and tumour necrosis factor induced proliferation of B-chronic lymphocytic leukaemia. AB - There is now good evidence that tumour necrosis factor [TNF] stimulates DNA synthesis of B-chronic lymphocytic leukaemia (B-CLL) cells. The malignant clone produces TNF, and addition of exogenous TNF up-regulates the TNF mRNA in B-CLL cells. Interleukin-6 (rIL-6) may also be important in this growth loop. We studied the interaction of TNF and IL-6 in the regulation of DNA synthesis (3H TdR uptake), cytokine release and cell survival in CLL cells in vitro. Addition of TNF (100 U/ml over 5 days) enhanced DNA synthesis from 718 +/- 284 (mean cpm +/- SE) to 2730 +/- 545 compared to cells cultured in medium alone (n = 16, p < 0.01). TNF-alpha induced DNA synthesis was inhibited in all cases studied by the addition of anti-TNF monoclonal antibody (5 micrograms/ml) to cell cultures. Spontaneous IL-6 protein release was enhanced in the presence of TNF (100 U/ml and 250 U/ml) by CLL cells at 48 hours of culture 143.6% and 172% (p < 0.05, n = 6). At 120 hours of culture, the increase was 323% and 412.5% (4 of 7 cases) of the control respectively. IL-6 (100 U/ml or greater) increased spontaneous DNA synthesis (3H-TdR uptake) but, in the presence of high concentrations of TNF alpha, inhibited TNF induced DNA synthesis in a dose dependent manner. Cell survival was reduced in the presence of anti-IL-6 mAb, while IL-6 was able to protect CLL cells from spontaneous apoptosis. These results suggest that IL-6 in an autocrine manner may inhibit DNA synthesis but prolongs survival in CLL cells. Increased serum IL-6 levels were detected in 27 of 50 cases of CLL, the mean level being significantly higher in Rai Stage III and IV cases compared to Rai Stage O-II cases. PMID- 8724533 TI - Morphologic evidence of apoptosis in childhood acute myeloblastic leukemia treated with high-dose methylprednisolone. AB - We have previously demonstrated that various subtypes of AML children respond to high-dose methylprednisolone (HDMP; 20-30 mg/kg/day) which could induce in vivo differentiation of myeloid leukemic cells to mature granulocytes. In this study we have evaluated whether apoptosis occurs in AML cells of patients treated by HDMP using morphological criteria. For light and electron microscopic examination bone marrow aspirates were obtained four days and two weeks after methylprednisolone (30 mg/kg/day) treatment from two children with newly diagnosed AML (AML-M3 and AML-M4). In both patients maturation of leukemic cells has previously been reported four days (in patient with AML-M3) and two weeks (in patient with AML-M4) after HDMP treatment. Electron microscopy revealed the characteristic ultrastructural changes of various stages of apoptosis four days after HDMP treatment in a case with AML-M3. Morphologic evidence of apoptosis induced by HDMP were also detected on Wright-stained and toluidine blue stained semithin sections of BM preparations in a patient with AML-M4 and AML-M3 respectively. These findings suggest that HDMP which could induce in vivo terminal differentiation in myeloid leukemic cells is also able to induce apoptosis in patients with AML. The possibility of HDMP-induced apoptosis should be evaluated in a larger series of patients with AML and other types of malignant tumors. PMID- 8724534 TI - Differentiating agents do not induce a true hairy cell phenotype in B-CLL cells in vitro. AB - B-Chronic lymphocytic leukemia (B-CLL) and hairy cell leukemia (HCL) are both differentiated B-cell lymphoproliferative disorders. Prior studies have suggested that phorbol esters and the macrocyclic lactone Bryostatin-1, which are both protein kinase-C activators, can induce the differentiation of B-CLL cells into HCL cells in vitro, as evidenced by morphology, phenotype and TRAP activity. The differentiating effect of all-trans retinoic acid on B-CLL cells has been less extensively studied. We studied the effects of incubating adherence purified B CLL cells with phorbol myristic acetate (PMA), all-trans retinoic acid (ATRA), and Bryostatin-1. None of these agents induced a true HCL phenotype (CD5-, CD11c/CD25 coexpression) under the conditions studied. PMID- 8724535 TI - Second malignancy in hairy cell leukaemia: no evidence of increased incidence after treatment with interferon alpha. AB - The last decade has seen a dramatic improvement in prognosis of hairy cell leukaemia (HCL) but concern has emerged with regard to the incidence of second malignancy. A recent report found an 18.8% incidence of second cancers and suggested a possible role for alpha-interferon (IFN) in their pathogenesis. We reviewed our larger series of 200 patients with HCL. We found second malignancies in 8 cases (4.0%), all but one of whom had received IFN. However, when compared to age- and sex- matched population data this represents no increase in relative risk of second cancer in patients with HCL and provides no evidence of a role for IFN in the pathogenesis of these second malignancies. PMID- 8724536 TI - 2-chlorodeoxyadenosine (2-CdA) in 2-hour versus 24-hour intravenous infusion in the treatment of patients with hairy cell leukemia. AB - Forty one patients with hairy cell leukemia (HCL) were treated with 2-chloro deoxyadenosine (2-CdA) administered in various schedules. Complete remission (CR) was achieved in 31 (76%) patients and partial remission (PR) in 9 (22%). The mean duration of remission (CR + PR) was 25.2 months (range 9-45 months). One patient did not respond to therapy. Twelve out of 16 patients (75%) achieved CR after 5 day intravenous infusions of 2-CdA and 19 out of 25 patients (76%) after 7-day courses. In 19 out of 23 patients (82.6%) CR was achieved after intermittent 2 hour infusions and in 12 out of 18 (66.7%) after continuous 24-hour infusion. The differences were not statistically significant. Side effects of 2-CdA were similar in both groups except for infections, which were less frequently observed in the group treated for 5 days. The results of our study suggest that 2-CdA can be effectively administered to patients with HCL using 5-day courses and a 2-hour daily infusion. PMID- 8724537 TI - Allele imbalance at tumour suppressor loci during the indolent phase of follicle centre cell lymphoma. AB - We have examined 41 cases of follicle centre cell lymphoma with fluorescent PCR of microsatellite repeats closely linked to or within six tumour suppressor gene loci (APC, DCC, P53, RB1, WT1 and NM23). These probes are highly informative with heterozygousity rates in the range of 57%-90%. In addition we have used four loci from chromosome 6 (D6S260, TNFa, D6S281 and D6S262) as control loci which are unlikely to be involved in the pathogenesis of lymphoma. Of 369 informative PCR reactions allele imbalance was identified in 38 (10%) and this was seen in 23 of the 41 cases. Looking at individual loci allele imbalance was seen in APC(1) 11%, APC(2) 12%, P53(1) 5%, P53 (2) 7%, WT1 5%, RB1 13%, DCC 18% and NM23 0%. This frequency of change was no different from that seen at the control loci D6S260 16%, TNFa 20%, D6S281 4% and D6S262 9%. In the indolent phase of germinal centre cell lymphoma there is therefore quite a high rate of allele imbalance at all loci but this is no higher in those loci linked to tumour suppressor genes. PMID- 8724538 TI - Abortive mitoses and nuclear DNA fragmentation in CD30+ large cells of Hodgkin's disease. AB - This study was undertaken to better comprehend the reasons for the scarcity of Hodgkin and Reed-Sternberg (H-RS) cells in Hodgkin's disease (HD) despite their expression of "proliferation-associated antigens". To this end, we assessed the relative frequency of mitotic phases and nuclear damage (detected by in situ end labeling of DNA strand breaks) in CD30+ large cells of nodular sclerosis and mixed cellularity HD. Our results show that a) most CD30+ cells in HD exhibit abortive mitoses, with a highly significant arrest at the metaphase-ana/telophase transition, and b) many of these elements, i.e. mainly H-RS cells, show fragmentation of nuclear DNA, suggesting imminent or actual death. Percentages of CD30+ cells that entered mitosis and those with DNA strand breaks were of a similar order of magnitude and correlated significantly in a linear fashion. These findings are consistent with the concept that cell deletion is the major cause of the paucity of H-RS cells in HD. PMID- 8724539 TI - Clonality switch in acute myeloid leukemia. AB - We have previously described a case of clonality switch in a female patient with acute myeloid leukemia (AML) by X-chromosome inactivation analysis. She presented with refractory anemia with excess blasts in transformation but soon progressed to overt AML. Following induction chemotherapy, she went into complete remission but later relapsed into a second myelodysplastic phase. Analysis of her X-linked DNA polymorphism patterns at presentation and relapse showed that hematopoiesis was clonal, but the genotypes of the two clones was different. She remains clinically well and has a virtually normal blood count more than 5 years from presentation. We now report an update of this unique case and discuss the implications of this finding within the context of a multicellular origin of leukemia. PMID- 8724540 TI - Monitoring bulky mediastinal disease with gallium-67, CT-scan and magnetic resonance imaging in Hodgkin's disease and high-grade non-Hodgkin's lymphoma. AB - Treatment of both Hodgkin's disease (HD) and high-grade non-Hodgkin's lymphoma (HG-NHL) with bulky presentation at diagnosis frequently results in residual masses detected radiologically. Conventional diagnostic radiology and computed tomography (CT) are generally unable to detect the differences between tumor tissue and fibrosis. Gallium-67-citrate (67Ga) SPECT and magnetic resonance imaging (MRI) can potentially differentiate residual active tumor tissue and fibrosis. Thirty-three patients with HD or HG-NHL presenting with bulky mediastinal disease were studied with CT, 67Ga SPECT, and MRI (only for 16 patients) at diagnosis, after two-thirds of their chemotherapy, at the end of chemotherapy, and after radiotherapy in order to evaluate the mediastinal region on the basis of persistence of residual masses and activity of pathological tissue. After treatment, all patients with 67Ga-negative (30/33) disease are still in continuous complete response. Among the three 67Ga-positive patients, 2 relapsed within one year and another one is still alive without evidence of disease. Regarding MRI, two patients were found to be positive, one of them concomitant with 67Ga-positivity; both patients survive in complete response. In lymphoma patients with bulky mediastinal presentation, the 67Ga SPECT remains the preferable imaging technique for monitoring and differentiating the eventual active residual tumor. In combination, CT and 67Ga SPECT represent a suitable complete imaging approach to the radiological diagnosis which may be useful in these particular patients. MRI could probably be considered as a second-line method and from our data would be used only in selected cases because of the high cost, accessibility, and lower specificity as opposed to 67Ga SPECT in evaluating potentially active residual disease. PMID- 8724541 TI - CD5-expressing B-cell lymphomas/leukemias: relatively high frequency of CD5+ B cell lymphomas with an overall poor prognosis in Nagasaki Japan. AB - To characterize CD5+ B-cell neoplasms in Japan, where chronic lymphocytic leukemia (CLL) is rare and of different subtypes in comparison with Western countries, we collected 58 cases of CD5+ B-cell lymphomas/leukemias and analyzed their clinicopathologic features. According to the French-American-British (FAB) and standard histologic classification, the cases corresponded to small lymphocytic lymphoma (SLL, group I; n = 22, consisting of CLL, n = 10, CLL/PL, n = 3, and CLLmixed, n = 7); intermediate differentiated lymphoma/mantle cell lymphoma (IDL/MCL, group II, n = 18); and others with CD5-positive lymphomas (group III, n = 18). The CD5+ B-cell lymphomas showed morphologic and prognostic variability among the three groups. The clinical and immunophenotypic features were remarkably consistent in leukemic disease being seen in 73% of all cases, splenomegaly in 63%, and intense CD19, CD20, surface membrane immunogobulin M (SmIgM) or SmIgM and SmIgD, light-chain expression, and no CD10 expression. The median survival time of groups I, II, and III was 7.8, 3.3, and 0.8 years, respectively. These findings suggest that CD5 antigens may serve as valid markers for the prognosis and clinical features of B-cell lymphomas and that CD5+ B-cell lymphomas with an overall poor prognosis occurs at a relatively high frequency in Japan. This also suggests that a combination of immunophenotypic and morphologic features is of value for characterizing CD5+ B-cell neoplasms. PMID- 8724542 TI - Esterase isoenzyme profiles of 255 leukemia-lymphoma cell lines from all hematopoietic cell lineages. AB - Carboxylic esterase isoenzymes isolated from a panel of well-characterized continuous human leukemia-lymphoma cell lines were separated by isoelectric focusing. Typical isoenzyme patterns designated Mono 1/Mono 2 (for monocyte associated), My 1/My 2 (for myeloid or myeloma), Lym 1/Lym 2 (for lymphoid) and Und (for undifferentiated) could be reproducibly discerned. The Mono patterns contained one unique isoenzyme encoded by the monocyte-specific esterase gene. This comparative analysis of 255 leukemia-lymphoma cell lines covered the major cell lineage that are affected by hematological neoplasias. The results showed that (except for myelomas) lymphoid-derived malignancies, both leukemias and lymphomas, expressed primarily the Und and Lym esterase isoenzyme profiles. In contrast, myeloid leukemia cells and the related erythroid and megakaryocytic cell lines displayed mainly the My patterns. The Mono patterns were detected predominantly in monocyte-derived leukemias. As the B-lymphocytic hierarchy progresses from pre B-cells via B-cells to plasma cells, number and intensity of the isoenzymes increased as well from the Und pattern to the My isoenzyme profile. Hodgkin's disease and anaplastic large cell lymphoma lines displayed heterogenous isoenzyme profiles consistent with their heterogenous cellular origin. The present study using continuous leukemia-lymphoma cell lines as model systems provides a biochemical characterization of different hematopoietic cell lineages and stages of differentiation. PMID- 8724543 TI - Establishment and characterization of a novel human bone marrow stromal cell line, FS-1. AB - We have established a human stromal cell line derived from the bone marrow of a patient with chronic myelogenous leukemia in blast crisis. This cell line, designated FS-1, exhibits a fibroblastoid morphology and does not express any hematopoietic cell marker tested. FS-1 is negative for alpha-naphthyl acetate esterase, acetylated LDL, von Willebrand factor, and shows no phagocytosis. This cell line is positive for acid phosphatase, alkaline phosphatase, collagen types I, III, IV, and fibronectin. cDNA from FS-1 cells was subjected to amplification by the polymerase chain reaction to assess the constitutive expression of several cytokine genes. Transcripts for interleukin (IL)-6, IL-7, macrophage colony stimulating factor (M-CSF), and stem cell factor (SCF) were detected in FS-1 cells. IL-6 and SCF also were detected in the culture supernatants of FS-1 at a concentration of 95 pg/ml and 21.2 pg/ml, respectively. These data show that FS 1, established from a human bone marrow, is a stromal cell line which was not generated using transfection with SV40 T antigen. FS-1 cells may be useful in supporting human hematopoietic cells for experimental manipulation. PMID- 8724544 TI - Granulocyte-colony stimulating factor, granulocyte-macrophage colony stimulating factor and interleukin 4 induce differentiation in the U-937 human monocytic leukemia cell line. AB - We studied the effect of TPA, G-CSF, GM-CSF, conditioned medium from 5637 cells (CM5637) and IL-4 on U-937 cell line proliferation and differentiation. Flow cytometry analysis showed that the expression of the CD14 cell surface antigen, initially detected in 90% of the cells, decreased when the cells were cultured with either G-CSF, GM-CSF, CM5637, or IL-4. The CD11c expression only decreased by exposure to GM-CSF and IL-4. The cells also showed a decrease in alpha naphthylesterase (alpha-NAE) activity and an increase in peroxidase (Px) activity in the GM-CSF supplemented cultures. Remarkable changes in cell morphology were also observed. IL-4 induced morphologic features resembling histiocytic-like cells positive for the expression of alpha-NAE and negative for Px. GM-CSF induced cells with pseudopods, negative for alpha-NAE expression and positive for Px. TPA effect on U-937 cells was similar to that observed with GM-CSF. No proliferative response was detected with any of the factors assayed. These results suggest that GM-CSF and IL-4 can promote distinct changes in the differentiative pathway of U-937 cells, as evidenced by the marked morphological, immunological and cytochemical changes observed in the cell cultures. PMID- 8724545 TI - FLANG (fludarabine + cytosine arabinoside + novantrone + G-CSF) induces partial remission in lymphoid blast transformation of Ph+chronic myelogenous leukaemia. AB - The adenine nucleoside analogue, fludarabine phosphate, in combination with cytosine-arabinoside (Ara-C) and granulocyte-colony stimulating factor (G-CSF) (the so called FLAG regimen) has recently been shown to be effective in the treatment of poor-prognosis acute non-lymphoid leukaemia. We used this combination plus novantrone (FLANG regimen) in a case of Ph1+ chronic myeloid leukaemia (CML) unresponsive to interferon alpha that had progressed to an acute phase, after 3 months of treatment with 6-mercaptopurine and hydroxyurea. The patient was treated with two courses of fludarabine 30 mg/m2 (days 1-5) + Ara-C 2 g/m2 (days 1-5) + novantrone 5 mg/m2 (days 1-3) and G-CSF from day 0 to neutrophil recovery. After the first cycle of chemotherapy, bone marrow blasts decreased from 100% to less than 5% (clinical complete remission), with a progressive clearance of Ph1+ metaphases (from 100% to 12%). At the end of the second course, a progressive increase of blasts was observed again and karyotypic detection of Ph+ cells was also documented (from 12% to 42.9%). During this partial remission, the patient underwent an allogeneic bone marrow transplantation from an HLA matched identical brother. At the time of this report, he is still alive and well and in complete karyotypic remission. This partial therapeutic success was compared with the result obtained in another previously reported CML case: differences in the therapeutic efficacy of protocols employing fludarabine nucleosides and the type of blastic cells involved are discussed. PMID- 8724546 TI - Hemophagocytic syndrome associated with Bacteroides fragilis infection in a patient with acute monoblastic leukemia. AB - The HS has been associated with malignant hemopathies. We report here a case of HS related to Bacteroides fragilis during the course of acute monoblastic leukemia. Evolution was fatal despite remission of the leukemia process and response of the infection to appropriate antibiotic therapy. PMID- 8724547 TI - Marked bone marrow eosinophilia at the time of relapse of acute myeloblastic leukaemia in association with the appearance of translocation t(12;20)(q24;q11). AB - We report a case of acute myeloblastic leukaemia (AML), FAB type M2, with karyotype t(8;21)(q22;q22), who at the time of relapse showed marked eosinophilia of the bone marrow. Karyotype analysis showed the appearance of an additional clone t(8;21)(q22;q22),t(12;20)(q24;q11). To the best of our knowledge, marked eosinophilia has not been reported in association with this particular chromosomal translocation. PMID- 8724548 TI - Granulocytic sarcoma in acute promyelocytic leukemia. PMID- 8724550 TI - Epidemiologic aspects of circadian patterns of cardiovascular disease and triggers of acute cardiac events. AB - An appreciation of the existence of circadian variation in association with the onset of acute coronary disease and of the public health and clinical implications of this temporal nonrandom disease occurrence with possible precipitating factors has been gained only over the past decade. Data providing support for the existence of circadian variation in the onset of acute coronary heart disease and the role of acute triggers are reviewed. PMID- 8724551 TI - Circadian patterns and triggers of sudden cardiac death. AB - Sudden cardiac death and other acute cardiovascular events have been demonstrated to occur in certain temporal patterns. The study of these patterns may yield important clues to the pathophysiology of the disease process. Most studies of the timing of onset of sudden cardiac death have revealed a prominent midmorning peak, thought to be related to a surge in catecholamines associated with arising and assuming the upright posture, that is blunted or eliminated by beta blockers. In addition, some studies have also shown a secondary peak in late afternoon or early evening of uncertain cause. The development of third-generation implantable cardioverter defibrillators with memory capabilities offers a unique opportunity to accurately define event chronology. PMID- 8724552 TI - Triggers for sudden cardiac death in the athlete. AB - Sudden death on the athletic field is usually due to underlying cardiovascular disease. Coronary artery disease is most common in older athletes, and a variety of congenital cardiovascular malformations predominate in young competitive athletes. Of these lesions, the most common in North America is hypertrophic cardiomyopathy. A variety of coronary artery anomalies are next in frequency, with the most important being anomalous origin of left main coronary artery from the anterior sinus of Valsalva. PMID- 8724553 TI - Pathophysiology and inflammatory aspects of plaque rupture. AB - Atherosclerotic plaque disruption with superimposed thrombosis is the main cause of acute coronary events. At present it is not known specifically how various risk factors influence the development and configuration of coronary plaques. The risk of plaque disruption depends more on plaque composition and vulnerability (plaque type) than on the degree of stenosis (plaque size). Lipid accumulation and chronic inflammation, particularly macrophage activities, seem to influence the development and the stability of a plaque and thus its propensity to rupture. Macrophages have the ability to secrete proteolytic enzymes and often infiltrate the region of imminent plaque rupture, and disruption may be associated with a generalized inflammatory reaction. Further, plaque disruption tends to occur at points where the plaque surface is weakest, corresponding to locations where mechanical and hemodynamic forces have their highest impact. The risk of plaque disruption is a function of both plaque vulnerability (intrinsic disease) and rupture triggers (extrinsic forces). The former predisposes the plaque to rupture; the latter may precipitate rupture. The danger of vulnerable plaques, however, is not related to disruption as such but to the resulting thrombosis. The magnitude of this thrombotic response and the ensuing flow obstruction depends on the contents and quantity of exposed thrombogenic plaque material, the degree of pre-existing stenosis, and the systemic thrombotic tendency. The thrombotic response is a dynamic process that is decisive for the clinical presentation and the outcome. PMID- 8724554 TI - Role of plaque size and degree of stenosis in acute myocardial infarction. AB - Angiographically apparent coronary artery stenoses limit coronary flow, produce symptomatic ischemia, and can be targeted for revascularization. Severe stenoses are more likely to occlude than segments without significant stenoses. Coronary angiography underestimates the extent of coronary atherosclerosis. Arterial segments without severe stenoses are much more common, and their risk of occlusion is not zero. Thus, the majority of myocardial infarctions are due to occlusion of arteries that do not contain obstructive coronary stenoses. Consequently, coronary angiography is not able to accurately predict the site of a coronary artery occlusion that subsequently will produce myocardial infarction. PMID- 8724555 TI - Hemodynamic changes as triggers of cardiovascular events. AB - Although a large number of clinical studies during the past decade have documented that myocardial ischemia, acute myocardial infarction, and sudden cardiac death have definite circadian patterns, recently published work suggests that it is not necessarily a specific time of the day but the sleep/wake cycle and postural changes as well as the balance between triggers and protective factors that determine the onset of cardiovascular events. A clear understanding of the pathophysiologic processes responsible for ischemic events and the specific roles played by various triggers would be helpful in better defining the therapeutic strategies designed for the prevention and treatment of acute cardiac events. Because changes in hemodynamic parameters are of paramount importance in initiating an ischemic event, it is critical to review their role and relationship in the pathogenesis of acute coronary events. PMID- 8724556 TI - Hemostatic factors as triggers of cardiovascular events. AB - A growing body of evidence supports a role for hemostatic factors in triggering cardiovascular events. Fibrinogen has been identified as an independent cardiovascular risk factor that is as powerful a predictor as cholesterol. Factor VII, fibrinolytic potential, von Willebrand's factor, and platelet reactivity also have been linked to increased incidence of cardiovascular disease. Further characterization of these factors may lead to improved risk assessment and the development of new therapies for prevention. PMID- 8724557 TI - Practical implications of circadian variations in thrombolytic and antithrombotic activities. AB - It recently has been demonstrated that thrombolytic therapy has circadian pattern of efficacy, as assessed by the ability to rapidly provide coronary patency. A study of 692 patients receiving intravenous tPA and undergoing acute coronary arteriography demonstrated a substantial diurnal pattern in patency with a peak at 8:00 pm. The heightened tendency for a coronary artery to be opened in the evening correlates well with the substantial tendency demonstrated in the same study and in multiple other studies for coronary arteries to thrombose and cause myocardial infarction in the morning hours. Circadian variations have been defined for a number of hemostatic and physiologic factors that would predispose toward clotting in the late morning, and converse circadian patterns have been described for a number of factors associated with thrombolysis that would predispose towards enhanced fibrinolysis in the evening hours. Methods by which efficacy of lytic therapy potentially could be enhanced include development of tPA variants or adjunctive agents that eliminate the circadian nadirs of efficacy, modification of dosage or choice of lytic agent as a function of time of treatment, and selection between pharmacologic lysis and direct angioplasty as a function of time of day. PMID- 8724558 TI - Physical exertion as a trigger of myocardial infarction and sudden cardiac death. AB - The data reviewed in this article indicate that physical exertion can trigger the onset of nonfatal myocardial infarction and sudden cardiac death. In addition, it is clear that although the relative risk associated with heavy exertion may be high, the absolute risk is actually quite small. It also is clear that regular exercise reduces the risk of triggering of myocardial infarction and sudden cardiac death by isolated bouts of exertion. Thus, these data provide further support for encouragement of regular exercise, as recommended by the American Heart Association. Such a program is likely to lower the overall risk of myocardial infarction and sudden cardiac death because it may lower the baseline risk and also decrease the relative risk that an episode of exertion will trigger a myocardial infarction or sudden cardiac death. Specific recommendations for patients with a history of myocardial infarction or angina are complex. Patients with coronary artery disease have the same relative risk of myocardial infarction and sudden cardiac death as those with no such history. Because of their elevated and variable baseline risk, however, specific recommendations regarding the risks and benefits of heavy physical exertion must be provided by their individual physicians, acting on recommended guidelines for exercise in such patients. PMID- 8724559 TI - Mental stress as a trigger of myocardial ischemia and infarction. AB - Recent research on the effects of behavioral activities on myocardial ischemia in coronary artery disease patients has provided a pathophysiologic model for understanding the mechanisms by which mental stress can trigger clinical cardiovascular events. This article reviews epidemiologic research implicating psychosocial stress as an acute trigger of myocardial infarction in patients with pre-existing coronary artery disease, and evidence for the pathophysiologic effects of acute mental stress in individuals with pre-existing coronary artery disease. Via its actions on the central and autonomic nervous systems, stress can produce a cascade of physiologic responses in vulnerable individuals that may lead to myocardial ischemia, ventricular fibrillation, plaque rupture, or coronary thrombosis. Also reviewed are field and laboratory studies that suggest important causal links between mental stress and myocardial ischemia, and evidence suggesting clinical significance for vulnerability to mental stress induced ischemia. PMID- 8724560 TI - Life-threatening cardiovascular consequences of anger in patients with coronary heart disease. AB - Anger is the affective state most commonly associated with myocardial ischemia and life-threatening arrhythmias. The scope of the problem is sizeable-at least 36,000 (2.4% of 1.5 million) heart attacks are precipitated annually in the United States by anger. The lethal cardiovascular consequences in ischemic heart disease are attributable to the unique physiology of this state, which activates high-gain central neurocircuitry and the sympathetic nervous system, leading to acute sinus tachycardia, hypertension, impaired myocardial perfusion, and a high degree of cardiac electrical instability. Exciting new tools have emerged from the fields of epidemiology, behavioral medicine, and cardiovascular physiology that offer considerable promise in accelerating our understanding of the pathophysiology of anger and in developing means to sever the link between anger and its life-threatening consequences. PMID- 8724561 TI - Prevention and practical aspects of triggering of cardiovascular events. AB - The recognition of the peak morning incidence of cardiovascular events and the role of triggering mechanisms can lead to some practical suggestions. For instance, adequate pharmacologic coverage over the full 24-hour period, in particular during the morning period of increased risk, is desirable among individuals taking antihypertensive and anti-ischemic medication. Although the absolute risk of an event occurring following a single exposure to a potential trigger is in general too low to recommend avoidance of the stressor, further research in this area may result in the design of pharmacologic and nonpharmacologic means to prevent the trigger from precipitating disease onset. PMID- 8724562 TI - How to perform cardiac MR imaging. AB - The complex motion of the beating heart provides a challenge to the clinical MR imager. Use of ECG gating, however, "stops" the heart, allowing for acquisition of diagnostic quality images of the cardiac chambers, valves, pericardium, and great arteries and veins and their branches. Attention to placement of chest wall ECG electrodes and the course of ECG-leads from the patient maximizes the quality of the gating signal and results in better quality imagery; however, ECG gating increases image acquisition time of the cardiac examination. Therefore, care must be taken to perform the most efficient possible examination. Prior to commencing the examination, the imager must have a clear understanding of the clinical question of the referring physician so that a clinical protocol can be applied to acquire relevant morphologic and physiologic data. The intracardiac anatomy may not always be best demonstrated using standard axial, coronal, and sagittal image sections. Therefore, with an understanding of basic intracardiac anatomy, standard imaging planes parallel or orthogonal to the intrinsic cardiac axes may be constructed and observation of abnormalities better appreciated. If care is taken in the planning and execution of the cardiac MR examination, the radiologist will be able to exploit this exciting new technology to its full potential as a noninvasive cardiac imaging modality. PMID- 8724563 TI - MR imaging of the thoracic aorta. AB - The evaluation and diagnosis of thoracic aortic disease using magnetic resonance imaging, including magnetic resonance angiography, is assuming an increasing role in clinical practice. Comparison with other modalities, including computed tomography and echocardiography, is discussed. Newer magnetic resonance imaging techniques, including 3-D breath-hold time-of-flight sequences are highlighted, as well as imaging strategies using standard magnetic resonance imaging techniques. Thoracic aortic disease entities covered include aneurysm, dissection, penetrating ulcers and intramural hematoma, aortitis and infection, tumors, and congenital aortic disease. PMID- 8724564 TI - MR evaluation of the pericardium and cardiac malignancies. AB - The pericardium and cardiac malignancies are well visualized on MR imaging because of the superb contrast resolution and multiplanar capability of the technique. This article highlights some of the clinical uses of MR imaging in evaluating the pericardium and cardiac lesions and provides comparison with echocardiography and CT scan. Specific diseases that are described include pericardial effusion, constrictive pericarditis, neoplastic pericardial disease, congenital abnormalities of the pericardium, benign, malignant and metastatic cardiac tumors, and paracardiac masses. This article discusses the impact of newer MR techniques such as cine MR imaging on the assessment of pericardial disease and cardiac lesions. PMID- 8724565 TI - MR imaging of acquired heart disease. AB - Spin echo and gradient reversal MR imaging techniques provide a noninvasive means of analyzing the state of ventricular myocardium in patients with atherosclerotic and valvular heart disease. Local ischemic changes are manifested on spin echo MR examination as loci of increased signal intensity, most likely representing accumulation of local myocardial edema. Administration of intravenous paramagnetic contrast material further enhances local changes, increasing the sensitivity of MR methods for detecting such change. Application of spin echo and gradient reversal techniques also provides direct demonstration of morphologic changes in chamber volume and myocardial wall thickness and thickening, the sequelae of ischemic and acquired valvular heart disease, as well as direct demonstration of abnormal blood flow across diseased valves. Newer methods of analysis of these flow abnormalities provide information concerning pressure gradients across stenotic valves and regurgitant fractions across insufficient valves. Thus, application of MR imaging techniques provides clinically relevant information needed by referring clinicians for the management of patients with acquired heart disease. PMID- 8724566 TI - MR imaging of cardiomyopathy. AB - Spin echo and cine MR imaging provides reliable and reproducible quantitative information about the hearts of patients with cardiomyopathy. In particular, these techniques are useful in demonstrating the severity and distribution of left and right ventricular myocardial thickness, changes in myocardial thickness during ventricular contraction, ventricular chamber volume, and change in volume through the cardiac cycle. Furthermore, direct examination of the ventricular myocardium by spin echo MR imaging is beginning to demonstrate its utility in characterizing abnormal myocardium and in providing inferential information concerning the nature of the myocardial abnormality itself. MR examination is also useful in excluding other causes of myocardial dysfunction that mimic cardiomyopathy as well as in helping define the type of cardiomyopathy, both of which allow better clinical management of these patients. PMID- 8724567 TI - Quantitative MR imaging of the heart. AB - MR imaging cardiac quantitation is accurate, highly reproducible, and feasible using equipment available in most hospitals. For determination of myocardial mass, evaluation of RV function, and quantitative measurements of flow in the great vessels and peripheral vasculature, MR imaging is the reference method. Availability of methods for rapid scanning and analysis will lead to increased use of cardiac MR imaging in quantifying cardiac function. PMID- 8724568 TI - MR imaging of pulmonary hypertension and right ventricular dysfunction. AB - Right ventricular cardiac function is altered by abnormalities affecting primarily the left-sided cardiac structures, the lungs, or the right-sided cardiac structures themselves. The most common cardiac causes for right ventricular dysfunction are chronic left ventricular ischemia and rheumatic mitral valvular disease. Pulmonary diseases that result in right ventricular dysfunction include pulmonary air-space disease, including emphysema, and pulmonary interstitial and parenchymal diseases, including idiopathic pulmonary fibrosis and cystic fibrosis. Chronic pulmonary vascular disease, including chronic thromboembolism and PPH have a significant effect on right ventricular performance. Common to all of these diseases is elevation of pulmonary vascular resistance with a commensurate increase in right ventricular pressure, resulting in right ventricular hypertrophy. The limited ability of right ventricular myocardium to function in the face of increased pulmonary resistance results in right ventricular dilatation, tricuspid regurgitation, and ultimately right ventricular failure. MR imaging provides direct, noninvasive visualization of the right ventricular chamber as well as the myocardium itself, allowing reliable demonstration of morphologic changes in the size and shape of the ventricle, thickness of the myocardium, and presence of abnormal infiltration by fat or edema. Furthermore, because MR imaging techniques do not depend upon geometric assumptions about the complex shape of the right ventricle, they may be used for accurate and reproducible quantitation of right ventricular volume and myocardial mass. PMID- 8724569 TI - MR imaging of congenital heart disease. AB - Patients with congenital heart disease often require mild sedation for the performance of their examination. If adequate monitoring of systemic oxygen saturation and heart rate is performed, then these patients may be adequately studied by MR imaging. Cardiac examination using spin echo pulse sequences provides morphologic information that allows evaluation of situs, ventricular morphology, and atrioventricular and ventriculoarterial connection. Gradient reversal acquisitions display changes in the size and shape of the atria and ventricles as well as demonstration of intracardiac and extracardiac shunts and abnormal flow across the cardiac valves. This information allows evaluation of the underlying congenital abnormality as well as the pathologic sequelae of the cardiac dysmorphology. Cardiac MR examination is a useful means of minimally invasive diagnosis of congenital heart disease and evaluation of its surgical palliation. PMID- 8724570 TI - Coronary MR angiography. AB - The principles underlying coronary MR angiographic pulse sequences are reviewed, including basic coronary anatomy, motion, and flow physiology. Coronary MR angiographic techniques can be subdivided into breath-hold (single or repeated) and nonbreath-hold techniques. The recent introduction of navigator pulses for real-time respiratory gating or triggering allows nonbreath-hold or repeated breath-hold 3-D coronary MR angiography, and will allow a more widespread use of this technique. PMID- 8724571 TI - Very fast cardiac imaging. AB - With very fast MR imaging techniques, myocardial perfusion can be assessed by monitoring the changes of myocardial signal intensity over time following bolus injection of a contrast medium. Very fast MR imaging techniques also allow for freezing of cardiac motion that is important for coronary MR angiography and their high temporal resolution allow for coronary flow reserve measurements. In the current overview, basic principles and strategies of very fast cardiac MR imaging are presented. In the second part, applications of these fast sequences in the fields of myocardial perfusion, coronary MR angiography, and low measurements are presented. PMID- 8724572 TI - Anatomy, function, and pathophysiology of peripheral nerves and nerve compression. AB - The basic pathophysiology of nerve compression injuries is complex and it is important to consider the microanatomy of the neuron and the peripheral nerve. The clinical stages of nerve compression lesions can be related to changes in intraneural microcirculation and nerve fiber structure; alterations in vascular permeability, with subsequent formation of edema; and deterioration of nerve function observed in experimental studies. The double-crush and reversed double crush syndromes are related to disturbances in axonal transport induced by compression, followed by morphologic and functional changes in the nerve cell bodies. An underlying neuropathy in subjects can make the peripheral nerves more susceptible to compression injuries. PMID- 8724573 TI - Biomechanical properties of peripheral nerves. AB - The biomechanical properties of peripheral nerves still are not completely understood or quantified. This is partly because of the nonhomogeneous nature of nerves and the variation in the proportions of the components of different nerves. The recognition and quantification of the in-situ stress and the viscoelastic behavior of nerves have been important stepping stones in understanding their mechanical behavior. The effects of stretch and compression on the vascular supply and conduction properties of normal animal nerves have been well studied. Similar studies in injured nerves will provide invaluable information for understanding the mechanics of nerve function and the prevention of stretch- and compression-related trauma. PMID- 8724574 TI - General principles and use of electrodiagnostic studies in carpal and cubital tunnel syndromes. With special, attention to pitfalls and interpretation. AB - Electrodiagnostic studies can help confirm the clinical impression of CTS and cubital tunnel syndrome with a high degree of sensitivity and specificity. These tests can provide valuable information regarding the exact point of nerve compression as well as the severity of damage. Differentiating these nerve entrapment syndromes from other conditions with similar clinical presentations can lead to effective therapy and can help predict response to treatment. The electrodiagnostic laboratory should employ the most current and sensitive techniques for obtaining optimal results, and the ordering physician should have a working knowledge of the principle involved in the tests to make best use of the resulting information. PMID- 8724575 TI - The role of splinting and rehabilitation in the treatment of carpal and cubital tunnel syndromes. AB - Successful treatment of patients suffering from carpal or cubital tunnel syndrome requires a skilled collaborative effort by the physician, therapist, and patient. Unfortunately, many factors influence an individual's recovery and outcome. As medical professionals, we must educate the patient, examine the physical and environmental factors involved in each case, and design unique treatment plans in each instance. PMID- 8724576 TI - The pathophysiology of carpal tunnel syndrome. AB - Carpal tunnel syndrome (CTS) is the most frequently encountered peripheral compressive neuropathy. Although the clinical diagnosis and treatment of CTS have been well defined, the pathophysiologic basis still is not understood completely. This article summarizes current thought and research pertinent to the pathophysiology of CTS. PMID- 8724578 TI - Anatomy of the median nerve at the wrist. Open carpal tunnel release--classic. AB - An understanding of the normal anatomy of the carpal tunnel and a variety of associated anomalies is important for the physician treating carpal tunnel syndrome. There are many strong arguments for open surgical decompression of the median nerve where full visualization of the transverse carpal ligament and contents of the carpal tunnel can be expected. The authors describe their preferred operative technique and post-operative management for carpal tunnel release. PMID- 8724577 TI - Vitamin B6 (pyridoxine) therapy for carpal tunnel syndrome. AB - The literature at this time does not give convincing evidence for use of pyridoxine as the sole treatment when confronted with a patient with idiopathic CTS. It may be of value as an adjunct in conservative therapy through altered perception of pain and increased pain threshold. For patients not responsive to conservative therapy, surgical decompression of the carpal canal is the treatment of choice. PMID- 8724579 TI - Carpal tunnel release with a small palmar incision. AB - The use of carpal tunnel tome with a small palmar incision to release the transverse carpal ligament is discussed. The technique is fully illustrated, and the authors early clinical experience is reviewed. PMID- 8724580 TI - Endoscopic carpal tunnel release. Chow technique. AB - The first cases using the set of instruments designed by James C. Y. Chow, MD, of Mount Vernon, Illinois, were performed in September of 1987 by the developer. His was the first written introduction of the technique of endoscopically assisted carpal tunnel release published by a US author. This technique has been practiced widely for 5 years. Serious complications can occur but may be avoidable. PMID- 8724581 TI - Endoscopic carpal tunnel release. The Agee technique. AB - The authors describe their technique for endoscopic carpal tunnel release using the Agree device. Technical points, illustrations, and postoperative regimen are provided. PMID- 8724582 TI - Carpal tunnel syndrome in the work place. AB - The management of patients with the CTS that appears to be related to occupational tasks is a complex issue. At this time, there is no definite evidence to show that any job is the sole cause of an individual worker's symptom complex. Management of patients in this situation requires surgical restraint coupled with an understanding of the worker's overall lifestyle. The most effective outcomes for all concerned appear to occur when there is cooperation among the patient, the physician, and the employer or his or her representatives. The recent prospective study by MacDougal that attempted to correlate job classifications with surgical outcome may be a very positive step in this direction. PMID- 8724583 TI - Reoperation for carpal tunnel syndrome. AB - Although primary carpal tunnel release is usually successful, reoperation is needed in up to 3% of patients. Common indications of reoperation are previous incomplete surgery and postoperative fibrosis. Although most patients improve after reoperation, persistent systems are likely and failure is more frequent than after primary carpal tunnel surgery. Risk factors for failure following reoperation include the presence of an active Worker's Compensation claim, pain in the ulnar nerve distribution, and the absence of abnormality on preoperative EMG. PMID- 8724584 TI - Complications of nonoperative and operative treatment of carpal tunnel syndrome. AB - The popularity of treatment of carpal tunnel syndrome has resulted in a significant number of complications. A rational plan of therapy is available for each category of complications of carpal tunnel syndrome. This article presents 14 of the most frequently encountered complications of treatment. Prevention and management of these complications are discussed. PMID- 8724585 TI - Recalcitrant carpal tunnel. The hypothenar fat pad flap. AB - Open decompression of the median nerve generally is so effective that little is mentioned of the surgical treatment options for recalcitrant or unrelieved carpal tunnel syndrome. The hypothenar fat pad flap has been shown to be a reliable local source of well-vascularized adipose tissue that can be used for coverage of the median nerve during re-exploration of recurrent or persistent idiopathic carpal tunnel syndrome. The hypothenar fat pad flap is a technically simple procedure that allows the fat pad to be mobilized easily and placed across the palm as a barrier between the nerve and the radial leaf of the transverse carpal ligament, effectively preventing median nerve readherence. This flap hopefully will improve the tissue environment for the median nerve, permitting it to have normal excursion during wrist motion. Our results to date have been better than previously described for other techniques. We believe the hypothenar fat pad flap should be considered in the hand surgeon's armamentarium for recalcitrant idiopathic carpal tunnel syndrome. PMID- 8724586 TI - The abductor digiti minimi flap: its use in revision carpal tunnel surgery. AB - One of the few muscles or myocutaneous flaps in the hand available to cover soft tissue defects without compromising significantly the hand's function and appearance is the abductor digiti minimi muscle. A technique that harvests the abductor digiti minimi flap with a small skin island from the hypothenar eminence allows direct closure of the skin following ADM transposition and makes it possible to directly monitor the blood supply to the muscle. This skin island is much smaller than others previously reported. PMID- 8724587 TI - The use of dermal-fat grafts. AB - Recurrent carpal tunnel syndrome is a difficult problem facing hand surgeons today, and its incidence is rising. In selected cases, re-exploration, median nerve neurolysis, and dermal-fat grafting are beneficial. The surgeon should anticipate 30% to 50% absorption of the graft. PMID- 8724588 TI - Reconstruction of the transverse carpal ligament to restore median nerve gliding. The rationale of a new technique for revision of recurrent median nerve neuropathy. AB - Two clinical and four surgical steps can be taken to return gliding and nutritional balance to the median nerve after revision surgery for a recurrent median nerve neuropathy: Step One (Clinical): An accurate diagnosis of the median nerve traction neuropathy. Consider brachial plexopathy and terminal neuropathies. Step Two (Surgical): Comprehensive exploration and meso epineurolysis of the median nerve from the radial remnants of the divided TCL and fibrosed radial and ulnar bursae by resection of the bursae and flexor tenosynovectomy. Step Three (Surgical): Fasciotomy of the volar carpal ligament to lengthen the ulnar leaf of the TCL; permits visualization of the ulnar artery and neurolysis of the ulnar motor nerve. Step Four (Surgical): Mobilize the previously released radical leaf TCL and motor median nerve from the origin of the thenar muscles. Step Five (Surgical): Restore the inner gliding surface and strength of the flexor retinaculum of the hand and wrist by reconstruction of the TCL and the forearm antebrachial fascia. Step Six (Clinical): An immediate postoperative hand therapy program to restore gliding to the median nerve and flexor tendons. Protocols for upper extremity nerve gliding should be carried out under supervision at selected daily intervals. A prevailing brachial plexopathy requires positive supervised therapy for nerve gliding and posture control. PMID- 8724589 TI - The synovial flap as treatment of the recurrent carpal tunnel syndrome. AB - After an exact definition of the recurrent carpal tunnel syndrome, its treatment, as described in the literature, is presented. To cover the median nerve isolated from the extended scar tissue with a thin and well-circulated tissue, the author proposes and demonstrates the synovial flap. The follow-up of 27 cases operated since 1979 with a follow-up time between 1 month to 14 years shows 6 excellent, 16 good, 3 satisfactory and 2 bad results. PMID- 8724590 TI - The use of the palmaris brevis flap in recurrent carpal tunnel syndrome. AB - In failed cases of primary carpal tunnel release, the addition of the highly vascular palmaris brevis turnover flap as an adjunct to internal neurolysis discourages scarring and provides a nutrient bed for axonal regeneration. In fact, the palmaris muscle flap actually may suppress the growth of nerve fibers into the overlying scar. Experimentally, classic neuromas do not form when a severed nerve is placed in an innervated muscle, particularly those with minimal excursion, as is the case with the turned over palmaris cushion. Major advantages of the palmaris turnover muscle are (1) proximity of the muscle to the operative field, obviating the need for a distant graft source; (2) negligible functional motor loss as a result of forfeiting this muscle; (3) no separate scar is created in raising this flap; and (4) adequate padding of the subcutaneous fat is still left at the donor site to protect the ulnar neurovascular bundle. Although postoperative healing and rehabilitation time are relatively lengthened by the more extensive dissection, subjective assessments of pain relief and clinical improvements in grip strength, pinch strength, and sensory parameters justify the use of the palmaris brevis turnover flap in recalcitrant carpal tunnel cases. PMID- 8724591 TI - General principles of patient evaluation and nonoperative management of cubital syndrome. AB - Cubital tunnel syndrome is reported to be the second most common entrapment neuropathy in the upper extremity. We can now easily appreciate how ulnar nerve pathology can occur at the elbow in the absence of trauma. We can also appreciate the role of conservative management in the treatment of a condition that previously was thought to be a surgical disease. PMID- 8724592 TI - Modified in situ decompression of the ulnar nerve at the elbow. AB - Controversy continues to surround the treatment of ulnar nerve compression at the elbow in the cubital tunnel. By performing extensive segmental electrodiagnostic studies of the ulnar nerve through the cubital tunnel, accuracy of diagnosis is improved. In those patients with documented compression where the ulnar nerve enters the flexor carpi radialis, decompression of this area alone yields excellent results with rapid recovery. PMID- 8724593 TI - Cubital tunnel syndrome. Treatment by medial epicondylectomy. AB - In summary, medial epicondylectomy for cubital tunnel syndrome provides relief or improvement of symptoms in most patients, with minimal morbidity. If symptoms persist, an anterior transposition can still be performed. PMID- 8724594 TI - Subcutaneous transposition of the ulnar nerve for treatment of cubital tunnel syndrome. AB - Subcutaneous transposition of the ulnar nerve has been widely reported as a successful surgical treatment for ulnar neuropathy at the elbow attributable to a variety of causes. Accepted indications for anterior transposition include any anatomic lesion that interferes with or impinges on the nerve along its native course. This may include a tumor, ganglion, osteophyte, valgus deformity or instability, or subluxation of the nerve, as listed previously. The surgical technique of ASCT also was described thoroughly earlier in this article. Points that warrant emphasis include thorough decompression along the entire course of the nerve, an attempt to preserve the venous plexus that accompanies the nerve, identification and preservation of branches of the medial antebrachial cutaneous nerve, and resection of a 3 to 6 cm segment of the medial intramuscular septum. Poor prognostic indicators include age over 50 years; relatively advanced neuropathy, as noted by electrical evidence of demyelination; or aggravating medical conditions, such as diabetes or alcoholism. Complications include neuroma of the medial antebrachial cutaneous nerve and resubluxation posterior to the medial epicondyle. In cases of reoperation for recurrent or persistent symptoms, inadequate release, most commonly at the medial intramuscular septum, was sited as the cause of failure in over 90% of cases. In a few cases, compression was found at the site of a fasciodermal sling. The majority of complications therefore were technical in nature and probably could have been avoided by strict attention to basic principles. Controversy surrounds the appropriate treatment for approximately half of patients in whom no clearly definable cause can be found. These cases are either attributed to "repetitive strain" or lumped into the "idiopathic" category. The pathophysiology leading to neuropathy in these groups is poorly understood, so the rationale for choosing one surgical procedure over another remains somewhat obscure. In the absence of an anatomic lesion, proponents of in situ decompression believe transposition involves unnecessary dissection, with attendant risks of devasularization or injury to the nerve or surrounding structures. Advocates of ASCT point out that the nerve may be compressed at any of several points along its course, as outlined in Fig. 1. Unlike in situ decompression, therefore, a properly performed anterior transposition assures adequate decompression at all points along its course. Indications for subcutaneous versus submuscular transposition are even less clear. Some believe submuscular transposition should be performed for more severe neuropathy, when muscular atrophy is present. Other authors point out that thin patients will be susceptible to repeated minor trauma if the nerve is left in a subcutaneous position. Neither of these contentions is supported consistently by available published data. In most cases of failed subcutaneous transposition, submuscular transposition has been used as a salvage procedure simply to place the nerve in an unscarred bed. Answers to the unresolved issues await well designed studies. Nevertheless, there is ample cause for optimism given that adherence to basic principles has resulted in satisfactory results for 85% to 95% of patients regardless of the procedure chosen. PMID- 8724595 TI - Anterior intramuscular transposition of the ulnar nerve. AB - The surgical management of cubital tunnel syndrome is well documented in the literature. Anterior intramuscular transposition of the ulnar nerve is indicated for chronic cubital tunnel syndrome with symptoms refractory to conservative therapy. Prompt diagnosis is essential to yield excellent results. Extreme care must be exercised in the performance of anterior intramuscular transposition. The surgeon must know the details of medial epicondylar anatomy and pathophysiology, as well as all possible sites of potential nerve compression. The placement of the transposed nerve in an intramuscular bed requires that all fibrous septae are resected from the shallow trough created for the nerve to avoid scar formation. Postoperatively, the arm is immobilized for 3 weeks, after which range-of-motion exercises are begun. By the eighth postoperative week, most patients are able to resume their regular activities, including manual labor. Recurrence or persistence of symptoms postoperatively typically is traced to an inadequate decompression of the nerve. Common sites of persistent ulnar nerve compression include (1) the medial intermuscular septum, (2) the arcade of Struthers, (3) fibrous bands immediately proximal or distal to the cubital tunnel, (4) persistence or kinking at the arcuate ligament of Osborne, (5) Spinner's ligament or other fascial slings, and (6) incomplete anterior transposition. Anterior intramuscular transposition of the ulnar nerve is attractive for its relative ease of dissection, simplicity, reliability, and low morbidity. Transposition of the nerve into a shallow muscular trough deep only to the flexor-pronator fascia is a logical, effective, and consistently reliable method of treating cubital tunnel syndrome refractory to conservative management. PMID- 8724596 TI - Submuscular transposition of the ulnar nerve. AB - Anterior submuscular transposition by the Learmonth technique is more demanding technically than other procedures described to treat cubital tunnel syndrome. I generally reserve submuscular transposition for patients who have failed previous anterior subcutaneous transposition and those who are very thin, in which case the nerve may be prominent immediately beneath the skin, resulting in an area of uncomfortable sensitivity. In patients who are candidates for reoperation following failed anterior submuscular transposition, it is common to find an area of compression that was not released during the initial operation. Most importantly, failure to release the arcade of Struthers, the arcuate ligament, and the flexor carpi ulnaris muscle fascia; excise the medial intermuscular septum; or provide ample room for the ulnar nerve beneath the flexor-pronator muscles will result in failure of surgical treatment. Range-of-motion exercises and hand strengthening facilitate early return of function. PMID- 8724597 TI - Cubital tunnel surgery. Complications and treatment of failures. AB - Cubital tunnel surgery should be considered a failure if patients have no relief of their symptoms of if the symptoms recur shortly after the surgery. Choice of treatment should be based on careful examination and evaluation of patient expectations, general medical condition, level of activity, and duration and severity of symptoms. Failure of the initial procedure may be the result of inadequate release, instability, subluxation, inadvertent creation of a new site of compression, and intraoperative nerve injury. Certain clinical manifestations specify the cause for failure. A positive Tinel's sign, for example, may indicate the exact location of persistent nerve compression. A palpable mobile mass on the medial aspect of the elbow is consistent with recurrent subluxation. Localized point tenderness along the course of the incision may indicate a neuroma secondary to injury to the medial antebrachial cutaneous nerve. In all cases, it is imperative that the surgeon be familiar with all the possible anatomic sources of compression as well as the variations in the ulnar nerve and the medial antebrachial cutaneous nerve. Once operative failure has been determined, efforts should be directed at completely releasing the nerve through external neurolysis, eliminating any mechanical stretch, and releasing any sites of compression or kinking. Some improvement should be expected if the surgeon thoroughly understands the anatomy, chooses the appropriate revision technique based on patient history, and adheres to the technical details of the chosen revision technique. PMID- 8724598 TI - [Biological role of cellular metabolites--structural analogs of platelet activating factor]. AB - The review is concerned with cell metabolites - the platelet activating factor (PAF) and its choline-containing structural analogs. The biological activity of PAF and its analogs are considered. The influence of PAF and its analogs of membrane properties, cell receptors, transmembrane signalling (protein kinase C activation, Ca2+ release) as well as its participation in cell phospholipases A2, C and D activation are considered. The intracellular function of PAF and its analogs is discussed in terms of their influence on prostaglandin and leukotriene biosynthesis as well as with reference to the biological activity of 1-acyl-2 acetyl-sn-glycerols and lysophosphatidylcholine. The function of cell-released PAF and its analogs is considered in connection with their influence on plasma lipoproteins, inter-cell PAF synthesis and different mechanisms of PAF and PAF analog-induced cell activation including cell priming, cell desensitization and receptor-dependent effects. It is concluded that PAF is an intracellular and intercellular mediator, whereas 1-acyl-PAF is an intracellular lipid bioregulator. PMID- 8724599 TI - [Conformational changes of the platelet membrane glycoprotein IIb-IIIa complex stimulated by a monoclonal antibody to the N-terminal segment of glycoprotein IIIa]. AB - During platelet activation, the glycoprotein (GP) complex IIb-IIIa (alpha 11b beta 3-integrin) changes its conformation, resulting in binding of adhesive proteins of RGD containing an amino acid sequence as well as in expression of new ligand-induced binding sites (LIBS) on the GPIIb-IIIa molecule. Like its F(ab) fragments, the monoclonal antibody CRC54, whose epitope is located in the N terminal part of the GPIIIa molecule, binds to no more than 10% of GPIIb-IIIa on the resting platelet surface. However, the binding of CRC54 increases considerably during activation of platelets by thrombin, platelet adhesion on plastic, GPIIb-IIIa interaction with RGDS-peptide as well as during dissociation of the complex in the presence of EDTA. These finding suggest that CRC54 is specifically directed against the LIBS epitope on the GPIIIa molecule. This epitope differs from those of other known conformation-dependent antibodies against GPIIb-IIIa (LIBS1, LIBS6, PMI-1, pl55 and p180), since those antibodies did not block the CRC54 binding to GPIIb-IIIa on the surface of adhering platelets. Unlike whole platelets, the binding of GPIIb-IIIa from lysates of platelets treated with Triton X-100 with immobilized CRC54 did not depend on the presence of the RGDS peptide. Under these conditions another anti-LIBS-antibody, p180 specifically directed against GPIIb, preserved its ability to discriminate the RGDS-occupied and resting conformations of GPIIb-IIIA. CRC54 and its F(ab) fragments induced platelet aggregation in both platelet-enriched plasma and in suspensions of washed platelets. CRC54 also stimulated the binding to platelets of GPIIb-IIIa ligand fibrinogen, labelled with 125I as well as adhesion of 51Cr labelled platelets to immobilized ligands-fibrinogen and fibronectin. The CRC54 dependent aggregation was fully blocked by RGDS-peptide and antibody CRC64 inhibiting the GPIIb-IIIa binding to the ligands. However, the platelet activation inhibitor, prostaglandin EI, and the mixture of metabolic inhibitors, deoxyglucose-sodium azide, only party inhibited the CRC54-dependent aggregation. Incubation of platelets with CRC54 induced the binding to platelets of the anti GPIIb LIBS antibody p180 and of the anti-GPIIb-IIIa activation-dependent antibody p155. The binding of GPIIb-IIIa from lysates of CRC54-treated platelets with immobilized p180 and p155 was also several times as high as that of GPIIb-IIIa from control platelet lysates. The data obtained indicate that the GPIIb-IIIa transition to the active state and its interaction with ligands induces conformational changes in the N-terminal part of GPIIIa and that the CRC54 binding to the N-terminal part of GPIIIa stimulates conformational changes in GPIIb-IIIa, complex interaction with ligands and platelet aggregation. PMID- 8724600 TI - [Mediated participation of the opioid system in regulation of pain sensitivity by peptide fragments MP1 and MP2]. AB - The naloxone-dependent modulating effect of synthetic analogs of myelopeptides, MP1 and MP2, on pain sensitivity in mice was determined at doses 10(-13) and 10( 8) g/animal. Binding of the peptides to opioid receptors of mouse brain membranes was studied by the radioligand assay. For MP1, the displacement of [3H]DAGO (selective mu-agonist) by IC50 = 7.3 x 10(-5) M and [3H]DSLET (selective delta agonist) with IC50 = 7.0 x 10(-5) M. The data obtained suggest that the hypoalgesic effect of the peptides cannot be due to their direct interaction with opioid receptors. However, the naloxone dependence provides a possibility of mediated involvement of the opioidergic system in the realization of this effect. PMID- 8724601 TI - [Dose-dependent character of the effect of hydrocortisone on energy metabolism of rat thymocytes]. AB - The influence of the hydrocortisone (GC) different doses on rat's thymocytes energetic metabolism was investigated. It has been demonstrated that as the prolonged action of GC in vivo (20 micrograms/kg of mass during 6 d. daily) as the preincubation of thymocytes with 10 microM GC in vitro are cytotoxic, leading to absolute uncoupling of oxidative phosphorylation (OP) and decreasing the rate of DNP-stimulated respiration speed. The injection of GC (20 micrograms/kg mass) at once before 72 hours to killing resulted the OP uncoupling almost on 2,5 time. Short-time action of GC at the same dose before 3 hours to drilling and so the thymocytes preincubation with 1 microM GC stimulated the transport processes on the level of substrate oxidation without the OP uncoupling. Short-time action of GC in vivo caused the inhibition of respiration, if we used glucose as the oxidative substrate, and stimulated the respiration in a presence of pyruvate Na in incubation media. PMID- 8724602 TI - [Negative cooperativity of 6-phosphogluconate dehydrogenase in rat liver]. AB - The effect of various NADPH concentrations on the activity of rat liver 6 phosphogluconate dehydrogenase (EC 1.1.1.44) has been studied. The influence of NADPH concentrations on the enzyme cooperativity was observed. NADPH used at concentration up to 200 microM appeared to be a competitive inhibitor with respect to NADP+ without any cooperative effect towards the coenzyme (NADP+) binding. At high concentrations of NADPH (above 300 microM) the negative cooperativity displayed by the enzyme was confirmed by a significant decrease of the Hill coefficient for NADP(+)-from 1.1 +/- 0.2 down to 0.6 +/- 0.1 (p < 0.05). PMID- 8724603 TI - [Effect of sn-2-acetylated analogs of phosphatidalic acid on release and aggregation of human platelets]. AB - The influence of sn-2-acetylated analogs of phosphatidalic acid on the release reaction and aggregation of human platelets has been studied. It was found that phosphate-, phosphorylethanolamine- or phosphorylcholine-containing sn-2 acetylated analogs of phosphatidalic acid do not stimulate human platelet aggregation but inhibit the platelet-activating factor (PAF) induced platelet aggregation. The phosphatidalic acid analog, I-O-alk-1'-enyl-2-acetyl-sn-glycero 3-phosphate, stimulates human platelet release reaction measured by acridine orange release from human platelets in the same extent as does PAF. Other phosphatidalic acid analogs (1-O-alk-1'-enyl-2-acetyl-sn-glycero-3 phosphoethanolamine, 1-O-alk-1'-enyl-2-acetyl-sn-glycero-3-phosphocholine) inhibit acridine orange exocytosis from PAF-stimulated human platelets. It is concluded that the activity of sn-2-acetylated phosphatidalic acid analogs towards human platelets differs from those of corresponding ester and ether phosphatidalic acid analogs and, consequently, they may have another functional significance in the processes of platelet activation. PMID- 8724604 TI - [Characteristics of distribution of nonpolar amino acid residues in amino acid sequences of different groups of globular proteins]. AB - A method for quantitative characterization of nonpolar residue distribution in amino acid sequences of different groups of globular proteins related to two amino acid subgroups: leucine, phenylalanine, tyrosine, tryptophan; and alanine, valine and isoleucine, has been developed. The value of proposed index for analyzed molecules of myoglobins are within the range of 0.65-1.35, those for cytochromes c are 1.00-1.37, for RNAases are 0.51-1.32, while the mean values for each protein group approximate unity. PMID- 8724605 TI - [Reactions of decarboxylated and side transamination during interaction of glutamate decarboxylase from Escherichia coli with substrate analogs, modified through C3 and C4 atoms]. AB - The interaction of glutamate decarboxylase with the aspartate and glutamate analogues modified at C3 and C4 was studied. 3-Arsonoalanine, 3-phosphonoalanine, 2-amino-4-arsonobutyric acid, 2-amino-4-phosphonobutyric acid, a mixture of diastereoisomers of 4-(methylthio) glutamic acid and erythro-4-(methylthio) glutamic acid were shown to be poor substrates for the enzyme. Their decarboxylation was accompanied by transamination of the coenzyme (PLP) to pyridoxamine phosphate (PMP) which reversibly inactivated the enzyme. With arsonoalanine only part of PLP was converted into PMP and another part irreversibly formed a complex. 4-(Methylsulfonyl)-L-glutamic and 4 [(phenyl)(hydroxy)phosphoryl]-L-glutamic acids did not react with the glutamate decarboxylase. PMID- 8724606 TI - [Features of inhibiting butyrylcholinesterase and carboxylesterase hydrolysis with fluoranhydride esters of beta,beta-diphenylethylphosphonic acid]. AB - A new type of organophosphorus compounds-beta, beta-diphenylethylphosphonic acid fluoroanhydride esters-with various alkyl radicals (CH3, C2H5, C3H7, i-C3H7, C4H9, i-C4H9, C5H11, C6H13) and a phenyl radical (C6H5) have been tested as inhibitors of horse serum butyryl cholinesterase (EC 3.1.1.8) and two forms of reindeer liver carboxyl esterase (EC 3.1.1.1). All the tested compounds are strong irreversible inhibitors of butyryl cholinesterase and strong combined type inhibitors of carboxylesterase. The values of inhibitory constants have been found to depend on the structure of the alkyl radical in the inhibitor molecule. PMID- 8724607 TI - [Characteristics of Saccharomyces cerevisiae nuclear polyphosphatase activity]. AB - Saccharomyces cerevisiae nuclei possess a polyphosphatase activity which is insensitive to a number of inhibitors of ATPase and pyrophosphatase (PPase) activities of the same organelle. Heparin, an effective inhibitor of the nuclear polyphosphatase activity, does not alter either the ATPase and PPase activity. The nuclear polyphosphatase activity is optimal at pH 7.5. Bivalent metal cations stimulate this activity in the following order: Co2+ > Mg2+ > Zn2+ > Mn2+. However, the magnitude of the stimulating effect is much lower than that for the polyphosphatase activities from other organelles of the same yeast. The polyphosphatase activity is nearly the same for polyphosphates ranging from [symbol: see text] = 9 to [symbol: see text] = 208, but is 1.5 times higher for tripolyphosphate. The K(m) values for the hydrolysis of polyphosphates with chain lengths [symbol: see text] = 3, 15 and 208 are 100, 5 and 4.1 microM, respectively. The polyphosphatase activity differs in some properties from that of the cell envelope, cytosol and vacuoles of the same S. cerevisiae strain. PMID- 8724608 TI - [Effect of specific activation of phosphatidylcholine metabolism in hamster fibroblasts transformed by Rous sarcoma virus]. AB - Transformation of embryonic hamster fibroblasts by the Rous sarcoma virus results in sharp increase of the turnover rate of one of cellular phospholipids phosphatidylcholine. The decrease in the rate of virus-transformed cells (HETSR strain) during the monolayer formation is attended by additional activation of phosphatidylcholine turnover. A similar effect is observed after prolonged culturing of cells with dexamethasone. Addition of the tyrosine kinase inhibitor, genistein, to cells leads to selective inhibition of phosphatidylcholine synthesis without any effect of phosphoinositide synthesis. Immunoblotting analysis of p60-src, the product of the viral oncogen v-src related to the tyrosine kinase family failed to produce any significant changes in protein synthesis and activity during dexamethasone-induced inhibition of HETSR cell growth. The data obtained testify to selective activation of phosphatidylcholine metabolism in src-transformed cells which enhances with a decrease in the rate of cell growth. The presence in HETSR cells of p60-src whose synthesis is not controlled by dexamethasone may be responsible for increased phosphatidylcholine metabolism and sustaining cell growth under conditions of limited activity of growth-promoting compounds. PMID- 8724609 TI - [Expression of HIV-1 epitopes included in particles formed by human hepatitis B virus nucleocapsid protein]. AB - Hybrids of the core protein of hepatitis B virus (HBcAg) have been designed which carry N-terminal insertions of B- and T-cell epitopes of HIV-1 an immunodominant B-epitope from gp41, a T-cell epitope from p34 pol, and a cluster of B- and T cell epitopes from p17 gag. The hybrids have been synthesized using two expression systems-one based on the thermoinducible PR promoter of bacteriophage lambda and the other one based on phi 10 promoter of bacteriophage T7 with 3-5% and 7-14% yields, respectively. The hybrids have dual HBV and HIV-1 immunospecificity and are assembled into particles similar to those formed by the protein carrier HBcAg. Sandwich ELISA and immune electron microscopy revealed that HIV-1 epitopes are exposed on the surface of the particles. PMID- 8724610 TI - [Stimulating effect of sodium ions on Escherichia coli growth in the presence of protonophore uncoupler]. AB - The increase of Na+ concentration from the submillimolar level to 50 mM stimulates growth of the Escherichia coli cells in the presence of the protonophorous uncoupler CCCP. The half-maximal effects is observed at 1 mM Na+. The decrease of inorganic phosphate concentration from tens of millimoles to 0.5 mM inhibits cell growth under same conditions. PMID- 8724611 TI - [Major p50 protein of the somatic cell cytoplasmic mRNP: expression in Escherichia coli, isolation, and some properties of the recombinant protein]. AB - The major mRNP protein of rabbit reticulocytes, p50, belonging to the family of Y box transcription factors has been expressed in E. coli. The isolation procedure of the recombinant protein has been described. The recombinant protein is similar to the protein isolated from rabbit mRNPs in SDS-PAGE mobility and interaction with specific antibodies. Similar to the natural protein, the recombinant protein forms homooligomeric complexes with a molecular mass of about 800 kDa, binds to the alpha-globin RNA and double-stranded DNA containing the Y-box. Both proteins can be phosphorylated in vitro. PMID- 8724612 TI - [Unique conserved segment of the amino acid sequence of insulin-like proteins]. AB - Insulins and insulin-like proteins have been found to contain a specific conservative site of the amino acid sequence. This site allows a significant determination of relatedness of the unknown protein to the given protein family. PMID- 8724613 TI - Evaluation of the prethrombotic state in pregnancy and in women using oral contraceptives. AB - The purpose of this study was to determine the effects of pregnancy and oral contraceptive use on in vivo coagulation by measurement of plasma levels of prothrombin fragment 1 + 2 and thrombin antithrombin III complex. Fifty-four women with uncomplicated singleton gestations, 34 women using oral contraceptives and 45 non-pregnant controls were the subjects of this study. Levels of prothrombin fragment 1 + 2 and thrombin-antithrombin III complex were measured using enzyme linked immunoassays. In uncomplicated pregnancies, levels of prothrombin fragment 1 + 2 and thrombin antithrombin III complex increased significantly with advancing gestational age. Women using oral contraceptives had significantly higher levels of prothrombin fragment 1 + 2 and thrombin antithrombin III complex compared to non-pregnant controls. The significant increase in levels of prothrombin fragment 1 + 2 and thrombin-antithrombin III complex throughout gestation and in women using oral contraceptives provided evidence of thrombin activation during these clinical conditions. PMID- 8724614 TI - A comparison of contraceptive habits and pregnancy outcome at 19 years of age in two cohorts of Swedish women born 1962 and 1972. AB - The prevalence of contraception and pregnancy outcome in two representative samples of 19-year-old women resident in the city of Goteborg, born in 1962 (n = 596) and 1972 (n = 641), respectively, was assessed and compared using a postal questionnaire technique. The prevalence of different contraceptive techniques was as follows (62/72): Oral contraception (OC) alone 44%/35%, (p < 0.001); OC + condom 1%/12%, p < 0.001; intrauterine device 4%/1%, p < 0.001; condom only 11%/14%; depot gestagen 0%/0.3%; no contraception 40%/38%. The median duration of OC use was 15 and 14 months, respectively. Fear of OCs was the commonest reason given for cessation of OC in the 62 cohort and that contraception was no longer required in the 72 cohort. The proportion of women who gave the latter explanation for cessation of OC had increased (p < 0.001) from 10% in 1981 to 57% in 1991, indicating that young women in Sweden now tend not to continue with OCs when a relationship ends. Although the prevalence of cigarette smoking was reduced (p < 0.05 in the 72 cohort compared to the 62 cohort), there was still an over-representation of smokers in both cohorts among contraceptive users (p < 0.001) and in women who had been pregnant (p < 0.001) or undergone an abortion (p < 0.001). Pregnancies were reported by 11% of the women from the 62 cohort and by 13% from the 72 cohort. A greater proportion (p < 0.001) of pregnancies terminated in legal abortion in the 72 cohort (61%) compared to the 62 cohort (50%). The medical complication rate following legal abortion was lower (p < 0.05) in the 72 cohort compared to the 62 cohort. PMID- 8724615 TI - A follow-up study of first trimester induced abortions at hospitals and family planning clinics in Sichuan province, China. AB - Four-thousand women aged 18-40 underwent an early induced abortion at hospitals and family planning clinics in six counties in Sichuan province, China, between 1 July 1990 and 30 June 1991. The subjects were followed-up three times, on days 15, 90 and 180 after the operation. Information was obtained about their age, parity, contraceptive use, social behaviors, and gynecological and psychological characteristics before and after the abortion. The results indicate that induced abortion is safe when provided by medically trained personnel in health facilities such as hospitals or clinics. Depressive symptomatology was assessed by the CES-D and SCL-90 scales. The depressive symptoms declined over time. A substantial proportion of the abortions were to unmarried women or resulted from non-use of contraception or contraceptive failure, implying that the incidence of unintended pregnancies and induced abortions could be reduced by more effective and accessible contraception. PMID- 8724616 TI - Morphological observations of vas deferens occlusion by the percutaneous injection of medical polyurethane. AB - Histological observations of 20 occluded vas segments obstructed by the standard procedure of medical polyurethane-vas occlusion in 10 volunteers requesting vasectomy and of 20 vas segments with plugs removed from 10 vas-occluded men requesting vas reversal were carried out in order to elucidate the mechanism of vas occlusion by medical polyurethane (MPU). Twenty vas deferens in vasectomy group were ruptured, only a small amount of MPU elastomer remained within the vas lumen, most having leaked through to encircle the ruptured vas. Histomorphology of the removed vas segments from 10 vas-occluded men, who had undergone MPU-vas occlusion 4 years earlier, showed diffuse proliferation of connective tissue, fibrosis, or hyalinization of fibroplastic tissue and local infiltration of lymphocytes and macrophages. The vas lumen both proximal and distal to the plugs was completely blocked. Sperm granuloma, foreign body granuloma, proliferation of nerve fibers and local infiltration of lymphocytes were found in four vas occluded men with painful nodules. These results suggest that the contraceptive mechanism of MPU-vas occlusion could be the result of secondary obstruction due to tissue proliferation by MPU irritation after the rupture of the vas deferens. PMID- 8724617 TI - Misoprostol is as effective as gemeprost in termination of second trimester pregnancy when combined with mifepristone: a randomised comparative trial. AB - A prospective randomized study was conducted to compare the efficacy of misoprostol with gemeprost when combined with mifepristone for termination of second trimester pregnancy. Patients requesting termination of second trimester pregnancy were randomized into two groups. In both groups of patients, 200 mg of mifepristone was given 36 to 48 hours before the administration of prostaglandins. In Group 1, the women were given 400 micrograms of oral misoprostol every 3 hours up to 5 doses. In Group 2, patients were given 1 mg of vaginal gemeprost every 6 hours up to 4 doses. The main outcomes measured were the induction-abortion intervals (the interval between the first dose of prostaglandin and abortion) and the incidence of side effects. Altogether, 50 subjects were recruited with 25 women in each group. The mean age and parity of the women and the mean gestational age of the two groups of women were comparable. There was no significant difference in the median induction-abortion intervals (8.7 hours in Group 1 and 10.8 hours in Group 2) or the incidence of side effects between the two groups of patients. We conclude that misoprostol is as effective as gemeprost in termination of second trimester pregnancy when combined with mifepristone. PMID- 8724618 TI - Beliefs about Norplant implants among low income urban women. AB - This study documents the counseling experiences and beliefs about Norplant implants that are held by an ethnically diverse sample of low income clinic patients in the United States. Virtually all implant acceptors, but only about half of those choosing other contraceptives, receive information about the Norplant system during clinic-based counseling. Exposure to group counseling and to a film regarding Norplant implants are associated with high levels of knowledge among acceptors. Several important patterns emerge in comparing the beliefs of patients choosing the Norplant system with those of patients choosing other methods of birth control. Substantial proportions of the latter group adhere to several inaccurate beliefs about the method, associating its use with: long-term health problems, problems for future babies, future fertility problems, infection, and high cost. These outcomes in particular, as well as several others described, should be systematically addressed and reviewed in contraceptive counseling. PMID- 8724619 TI - An evaluation of the copper-T 380A IUD's safety and efficacy at three African centers. AB - The use of intrauterine devices in Africa is low compared with other contraceptive methods such as oral contraceptives. This study, coordinated by Family Health International, evaluated the clinical performance (safety and efficacy) of the TCu 380A IUD in three African centers, respectively, in Cameroon, Egypt, and Nigeria from 1986-1989. The 12-month unintended pregnancy rates were low for all three centers, ranging from none to 1.6 per 100 women. The 12-month discontinuation rates for all reasons ranged from 8.8 to 26.9 per 100 women. Statistically significant differences were observed among clinics for discontinuation rates for bleeding and/or pain and for planned pregnancy. The overall performance of the TCu 380A IUDs was considered satisfactory. Thus, the limited use of IUD in Africa could be due to the lack of accessibility, lack of information about, and confidence in the method rather than to documented clinical performance. PMID- 8724620 TI - Antifertility effects of an LHRH agonist in male mice. AB - This study was designed to investigate the effects of repeated high doses of an LHRH agonist on Swiss porton mouse seminiferous epithelium. Seminiferous epithelium showed more pronounced degenerative effects at the higher dose (1000 micrograms/kg wt/day) of the drug, with arrest of spermatogenesis at all stages of differentiation; effects on spermatogenesis become more evident after pachytene spermatocyte stage. The decrease in the testis-specific enzyme LDH-X is accounted for by the decrease in the number of cells of gametogenic origin. Although complete azoospermia was not observed, acute inhibition of motility of spermatozoa may account for severely decreased fertility rates after mating. However, reduced fertility rates due to decreased libido as a consequence of diminished testosterone levels cannot be discounted. PMID- 8724621 TI - Pharmacokinetics of ethinyloestradiol in rabbits after intravenous administration. AB - The pharmacokinetics of ethinyloestradiol (EE2) after intravenous administration of 30, 50 and 100 micrograms.kg-1 was investigated in rabbits. A high-performance liquid chromatographic (HPLC) method with electrochemical (EC) detection was used to measure EE2 in plasma samples in order to avoid the interferences of natural oestrogens. After compartmental analysis, the disposition of EE2 was well described by a two-compartmental open model with mean values of: alpha = 0.3448 +/- 0.2922, 0.1965 +/- 0.1755, 0.3058 +/- 0.1225 min-1, and beta = 0.0137 +/- 0.0018, 0.0140 +/- 0.0065, 0.0198 +/- 0.0066 min-1, for the three doses studied, respectively. There were no dose-related differences (ANOVA, P < 0.05) in a, b or Vss, but significant differences were detected in clearance (90.9 +/- 18.7; 80.6 +/- 17.6; 116.3 +/- 21.5 ml.min-1.kg-1) between the 100 micrograms.kg-1 group and lower dose groups. The AUC increased significantly with the doses (341.7 +/- 67.1; 645.8 +/- 143.9; 892.2 +/- 211.9 ng.min.ml-1). After non-compartmental analysis there were no significant differences in lambda, MRT or Vss as a function of dose, but these differences were significant when Cl or AUC were compared. There were no significant differences in AUC or Cl values obtained by compartmental and non-compartmental analysis. PMID- 8724622 TI - Assessment of the anti-microbial agent C31G as a spermicide: comparison with nonoxynol-9. AB - The broad-spectrum anti-microbial agent, C31G, containing an equimolar mixture of n-dodecyl-dimethylamine-N-oxide (C12-N-O) and N-(n-dodecyl), N-dimethyl-glycine (C12-betaine), was tested for spermicidal activity in comparison with the currently used spermicide, nonoxynol-9 (N-9). The rate of sperm cell permeabilization by the spermicides, as assayed with the fluorescent probe, TO PRO-1, increased as the cube of the C31G concentration, while the rate increase was linear with N-9 concentration. At 0.04%, the rate of sperm cell permeabilization with both spermicides is at the limit of rapid measurement. C31G diffuses through cervical mucus at a more rapid rate than does N-9. C31G has long been known to aid wound healing and reduce inflammation, whereas N-9 has been reported to induce vaginal irritation. C31G would, thus, seem to have the spermicidal efficacy, the broad range of anti-microbial activity, and the lack of inflammatory activity that is sought in the ideal vaginal spermicide. PMID- 8724623 TI - Molecular genetics of inversion 16 leukemia: implications for leukemogenesis. PMID- 8724624 TI - Deletion of chromosome 5 (5q-) in leukemia: current knowledge. PMID- 8724625 TI - Bcl-2 family proteins and the regulation of programmed cell death in leukemia and lymphoma. PMID- 8724626 TI - MLL fusion genes in the 11q23 acute leukemias. PMID- 8724627 TI - The c-myb proto-oncogene: a novel target for human gene therapy. PMID- 8724628 TI - The role of p53 in malignancy. PMID- 8724629 TI - Primitive hematopoietic stem and progenitor cells in human umbilical cord blood: an alternative source of transplantable cells. PMID- 8724630 TI - Detection of minimal residual disease in all: biology, methods, and applications. AB - The PCR technique appears to be the most sensitive method for detecting residual disease in ALL and can be applied to a high percentage of cases by amplifying sequences of the antigen-receptor genes. The PCR studies to date suggest that this sensitive technique can detect residual disease in virtually all patients during the first year of treatment. The residual disease becomes undetectable in the majority of patients by the end of treatment; however, a subset of patients remain PCR positive at a time when therapy is electively discontinued. The development of a highly accurate quantitative PCR technique may allow the possibility of distinguishing the patterns of residual disease for patients who will be cured by treatment from those who relapse. If such a pattern can be discerned, then an immediate benefit for PCR monitoring will be that clinicians will have the opportunity to test whether treating patients at the time of 'molecular relapse' will help to improve the cure rate for this disease. The PCR studies of remission marrows at the end of treatment raise a number of questions about the biology of disease persistence in patients who remain in extended 'remission.' A commitment to obtaining and analyzing bone marrow specimens in patients who have completed therapy is necessary to discern whether novel strategies, such as immunomodulatory manipulations, are needed to control or eradicated residual disease in patients who have completed planned chemotherapy. Thus, the long-term benefit of residual disease monitoring by PCR may be a better understanding of the biology and definition of 'cure' in ALL. PMID- 8724631 TI - Interleukin-6: biologic properties and role in lymphoproliferative disorders. PMID- 8724632 TI - Autologous bone marrow transplantation for leukemia and lymphoma. PMID- 8724633 TI - Retinoblastoma gene in malignancy. PMID- 8724634 TI - Treatment of relapse after allogeneic bone marrow transplantation. PMID- 8724635 TI - Acute lymphocytic leukemia: a comprehensive review with emphasis on biology and therapy. PMID- 8724636 TI - Plethysmography: safety, effectiveness, and clinical utility in diagnosing vascular disease. AB - Plethysmography, a semiquantitative method of measuring segmental blood flow and velocity in the carotid and peripheral vascular systems, is safe, easy to perform, and inexpensive. Impedance, strain guage, air, and photoelectric plethysmographic methods are assessed in this publication for their relative safety, efficacy, and clinical utility in diagnosing vascular disease. Clinical evidence has demonstrated, for example, that oculoplethysmography is not a reliable screening test for carotid artery disease, particularly in bilateral disease and nonocclusive unilateral disease. Oculoplethysmography is thus no longer recommended for the diagnosis of carotid artery disease. Impedance, strain guage, and photoplethysmography methods can be used for the initial evaluation of acute and chronic venous insufficiency, although there is great variability in the reliability of these tests to predict venous disease in the presence of nonobstructive thrombi and comorbid conditions. Strain guage and photoelectric plethysmography can be safely used for the evaluation of peripheral arterial disease. But surgical candidates for arterial reconstruction and some venous disorders may require additional imaging such as duplex ultrasound, venography, or arteriography for anatomic information that is not elucidated by plethysmography. The plethysmographic techniques reviewed in this assessment, evaluating the ability to predict the presence of flow reduction in the carotid and peripheral circulations, are based on the technical performance of the tests as compared with a reference method. Establishing evidence-based conclusions proved difficult in light of biases such as nonrandomization, unstated patient selection criteria, poor followup of patients, and lack of blindness in some studies. PMID- 8724637 TI - Infantile cataracts. AB - Cataracts are one of the most treatable causes of visual impairment during infancy. Recent epidemiological studies have shown that they have a prevalence of 1.2 to 6.0 cases per 10,000 infants. The morphology of infantile cataracts can be helpful in establishing their etiology and prognosis. Early surgery and optical correction have resulted in an improved outcome for infants with either unilateral or bilateral cataracts. While contact lenses continue to be the standard means of optically correcting an infant's eyes after cataract surgery, intraocular lenses are gaining in popularity as an alternative means of optically correcting these eyes. Post-operative complications occur more commonly after infantile than adult cataract surgery and many of these complications do not develop until years later. As a result, it is critical that children be followed closely on a long term basis after infantile cataract surgery. PMID- 8724638 TI - Risk management issues in glaucoma: diagnosis and treatment. AB - This article is based on the analysis of 194 claims in cases with a diagnosis of glaucoma. Half of these resulted in indemnity payments, and these payments were nearly 20% higher than the average paid for ophthalmology claims. Risk management guidelines are given. PMID- 8724639 TI - Treatment of dry eye by blocking the lacrimal canaliculi. AB - Occlusion of the lacrimal canaliculi improves the objective signs and subjective symptoms of dry eye. In this review, methods of occlusion are classified as surgical, thermal and tamponade. Surgical methods include dacryocystectomy, canalicular ligature, canalicular offset, canalicular excision, transfer of the punctum to dry dock, punctal tarsorrhaphy and punctal patch. Thermal methods include cautery, diathermy and laser burn. Tamponade methods use absorbable inserts of hydroxypropyl cellulose, gelatin, collagen and catgut, and nonabsorbable inserts of silicone (punctum plugs, canalicular plugs), polyethylene, cyanoacrylate, and others. The characteristics of all these methods are analyzed. PMID- 8724640 TI - Normal and pathological mechanisms in retinal vascular development. AB - Angiogenesis is a complex biologic process that occurs normally in development and in turnover and remodeling of mature vascular networks. Pathological angiogenesis and neovascularization occur in association with retinal and ocular ischemic diseases, in retinopathy of prematurity and other developmental disorders, and in tumor growth and metastasis. We describe current understanding of cellular and molecular mechanisms of retinal vascular development, highlighting aspects that relate to eye diseases, that provide sites of therapeutic intervention in ophthalmology and that are potential avenues for research. PMID- 8724641 TI - Success of monovision in presbyopes: review of the literature and potential applications to refractive surgery. AB - The monovision concept of correcting one eye for distance and the other for near may be utilized in presbyopes undergoing refractive surgery. We have performed a systematic review of published literature to evaluate the factors influencing monovision success, and to determine the visual outcome in patients with monovision. Articles in MEDLINE and published bibliographies reporting monovision prescription for correction of presbyopia were systematically identified and reviewed. Pertinent data were abstracted and, when feasible, statistically analyzed. The mean success rate was 73%. The success in monovision correlated with distance correction on dominant eye, alternating dominance, less than 50 seconds of are stereoacuity reduction, and less than 0.6 prism diopter of distance esophoric shifts. Monovision resulted in significant reduction of binocular contrast sensitivity function at spatial frequencies higher than 4 cycles per degree, and 2-6% reduction in task performance, but resulted in minimal reduction of binocular visual acuity, peripheral vision, visual field width and binocular depth of focus. The published literature indicates that monovision is an effective and reasonable therapeutic modality for correcting presbyopia. Proper patient selection and clinical screening are essential for monovision success. PMID- 8724642 TI - Ocular ischemic syndrome in a child with moyamoya disease and neurofibromatosis. AB - Ocular ischemic syndrome is extremely rare in childhood. Patients with moyamoya disease may be particularly susceptible to the development of ocular ischemia due to the associated carotid occlusion. A 19-month-old boy presented with neurofibromatosis and signs of ocular ischemia. At 29 months of age, he developed dense right vitreous hemorrhage and eventually lost vision in that eye due to phthisis. At almost six years of age, he developed an acute hemiplegia and was then diagnosed with moyamoya disease. This rare instance of childhood ocular ischemia in conjunction with moyamoya disease and neurofibromatosis demonstrates the serious ocular and systemic sequelae of occlusive vascular disease. PMID- 8724643 TI - Lung cancer with visual loss. AB - A 58-year-old man with primary large cell carcinoma of the lung presented with rapidly progressive, bilateral visual loss. The patient was alert and oriented, had no complaints of headache, and was found to have full ocular motility. The optic disks and fundi appeared normal. No visual pathway lesions or other CNS abnormalities were detected on neuro-imaging. The authors discuss their differential diagnosis, clinical diagnostic approach, and subsequent management of this unusual patient. PMID- 8724644 TI - Neurotoxic effects of DSP-4 on the noradrenergic system of the goldfish brain. AB - The substance N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine (DSP-4) is a neurotoxin with selective and long-lasting effects on the noradrenergic (NA) neurons of mammalian brains. The present study examines the effects of this toxin on the noradrenergic system of the goldfish brain. Single doses (50 mg/kg body weight) of DSP-4 reduce the immunoreactivity of the NA synthesizing enzyme dopamine-beta-hydroxylase (DBH), as revealed by immunohistochemistry 7 and 12 days after toxin administration. The depletion involves the DBH-positive fibres and spares the DBH-positive cell bodies. Dopamine-beta-hydroxylase immunoreactivity, 40 days after toxin administration, showed a complete recovery. Ultrastructural investigations confirmed that DSP-4 toxicity affects only nervous fibres and terminals, sparing cell bodies. Administration of DSP-4 also produced a marked decrease of noradrenaline (NA) levels in the goldfish brain, seven days later, while dopamine (DA) and serotonin (5-HT) levels were unaffected by toxin injection. The reduction of NA levels induced by DSP-4 was prevented by the concomitant administration of the NA uptake inhibitor desipramine. Noradrenaline levels measured 40 days after toxin administration show that DSP-4 toxicity was completely reversed. The results suggest a pronounced plasticity of the noradrenergic system in the goldfish brain. PMID- 8724645 TI - Does bigger mean better? Evolutionary determinants of brain size and structure. AB - Current perspectives on brain evolution relate brain size variability to two main parameters: a scaling factor that corresponds to overall body size and an ecological factor associated with behavioral capacity. I suggest in this paper that in evolution body weight and ecological conditions have different effects on brain structure, resulting in distinct differences in neural architecture, even if both factors may produce brain size increases. There are two postulated modalities of brain growth, one passive that lags behind increases in body size, and one active that relates to selection of specific behavioral abilities and hence increased processing capacity. These two modes of growth differ in three main aspects: (i) cellular and connectional rearrangements are modest in passive brain growth while they are conspicuous in active growth, corresponding to increases in processing capacity; (ii) passive brain growth follows a rather conservative allometric rule between brain components, while active growth usually affects only a few brain parts, thereby producing much steeper allometric relations between these parts and sometimes also in brain/body relations; and (iii) passive growth may either affect early periods of ontogenic brain development or produce a generalized increase in cell proliferation in later periods. On the other hand, active growth is restricted to relatively late developmental periods. Finally, an evolutionary scenario for the active mode is proposed where phylogenetic selection of an increased number of cells in particular brain regions occurs in order to facilitate neural reorganization and to increase the specificity of connections. This view emphasizes the role of connectional modifications in increasing brain capacity and contrasts with current ideas of a unitary process of phylogenetic brain growth, where a larger brain size per se produces better processing capacity, regardless of the causal factor behind it. PMID- 8724646 TI - Ontogeny of cholecystokinin binding sites in the hindbrain of the laboratory rat. AB - Studies in our laboratory have revealed a robust, transient expression of cholecystokinin binding sites in the facial motor nucleus during development in the Brazilian opossum, Monodelphis domestica. To investigate the ubiquity of this phenomenon, we have performed receptor autoradiography on the hindbrains of embryonic and neonatal rat pups. Cholecystokinin binding sites are present at very low levels in the embryonic day-16 rat hindbrain, but binding sites are abundant prior to birth. The greatest increase in labelled nuclei occurs prior to 5 days of postnatal age. Binding levels are heavy in the nucleus of the solitary tract, medial vestibular nucleus, posterior dorsal tegmental nucleus, area postrema, and caudal spinal trigeminal nucleus by 30 days postnatal. Both A-type and B-type receptors are present in the neonatal brainstem, although most labelled areas appear to be B-type. A-type binding sites are present in the ventral cochlear nucleus, the nucleus of the solitary tract, the dorsal motor nucleus of the vagus, the area postrema, the spinal nucleus of the trigeminal, and the cuneate and gracile nuclei by 5 days postnatal. As reported for the Brazilian opossum, cholecystokinin binding sites are expressed in the facial motor nucleus of neonatal rats and are transient. In this study of the brainstem in laboratory rats, a transient expression is also observed in the rubrospinal tract, parvocellular reticular nucleus, raphe obscurus, cuneate and gracile nuclei, and the ventral median fissure of the spinal cord. As vasopressin binding sites and estrogen receptors have also been shown to be expressed transiently in the laboratory rat facial motor nucleus, the physiological and developmental significance of transient binding site expression remains to be elucidated. PMID- 8724647 TI - Mitochondrial distribution in visual and auditory cerebral cortex of the harbour porpoise. AB - The distributions of mitochondria and synapses in two areas of harbour porpoise cerebral cortex were examined by quantitative electron microscopy of sections stained for cytochrome oxidase. The distribution of cytochrome oxidase-positive and total mitochondria in the visual cortex of the lateral gyrus and in the auditory cortex of the temporal operculum was related closely to that of total cytochrome oxidase staining seen by light microscopy in the relevant areas. There were two peaks of mitochondrial numerical density in visual cortex: in layer III and the upper part of layer I. Mitochondrial distribution was more uniform in temporal cortex, where the numbers of mitochondria in layers VI, V and lower I were similar to those in visual cortex, but fewer were present in layers III, II and upper I. The laminar distribution of axodendritic synapses in both cortices was relatively uniform, and there was not such a large difference between the two areas. As large numbers of mitochondria have been described in the layers of cat visual cortex showing dark staining for cytochrome oxidase and receiving thalamic afferent input, we regard our data as suggestive of the existence of two main thalamorecipient zones in cetacean cortex: one in layer III and the other in upper layer I. PMID- 8724648 TI - Organization of GnRH and FMRF-amide systems in two primitive bony fishes (order polypteriformes). AB - To investigate the evolution of the neural organization of gonadotropin-releasing hormone (GnRH), we have examined GnRH-immunoreactivity in two brachiopterygian fishes (Polypterus palmas and Calamoichthys calabaricus). Distal regions of the terminal nerve (TN) within the medial olfactory nerve contained clusters of GnRH immunoreactive (ir) perikarya (< 10 microns). More proximal, isolated GnRH-ir neurons were present among TN fascicles as they penetrated the ventral forebrain, and a few ir neurons were observed accompanying GnRH-ir fibers in the rostromedial telencephalon. GnRH-ir neurons were not observed in the preoptic area or ventral hypothalamus. In contrast, a small group of GnRH-ir neurons was localized in the periventricular nucleus of the posterior tuberculum. GnRH-ir fibers were present in widespread areas of the brain, including the olfactory bulb, telencephalon, optic nerve, hypothalamus, thalamus, habenula, optic tectum, tegmentum, pituitary and spinal cord. To further characterize projections of TN neurons, we utilized antiserum to FMRF-amide, a small peptide produced by TN cells in other vertebrates. Perikarya that were FMRF-amide-ir within the TN were similar in distribution to GnRH-TN neurons, and the distribution of FMRF-amide-ir fibers overlapped those of GnRH-ir fibers, thus providing a useful marker for identifying TN projections. An additional population of FMRF-amide-ir neurons was present in the periventricular hypothalamus. Our results suggest that in the polypteriformes, GnRH and FMRF-amide neurons of the TN are similar to those observed in other vertebrates; however, the paucity of GnRH cells in the basal forebrain may be unique to primitive actinopterygians and elasmobranchs, and may result from the lack of migration of GnRH neurons into the forebrain, a phenomenon that likely occurs in all other vertebrate classes. Finally, the identification of GnRH-ir neurons in the posterior tuberculum is consistent with similar, and perhaps homologous, GnRH neurons present in nearly all other vertebrate classes. PMID- 8724649 TI - Expression of complement regulatory proteins on fetal blood cells in utero. AB - Since the fetus is semiallogenic to the mother, mechanisms have evolved to protect fetal tissue from the maternal immune response. Among these mechanisms is the expression of cell-surface complement regulatory proteins at the maternal fetal interface. However, beginning in the third trimester, fetal blood cells are exposed to actively-transported IgG antibody. Thus, we speculated that fetal blood cells would require expression of one or more complement regulators by the early third trimester. Using flow cytometry and Western blots, we have demonstrated the presence of three important complement regulatory proteins in the circulating blood cells of human fetuses. These findings are consistent with the putative biological role of the cell-surface complement regulatory proteins. PMID- 8724650 TI - Effect of subadequate maternal vitamin-A status on placental transfer of retinol and beta-carotene to the human fetus. AB - The placental transfer of retinol and beta-carotene was assessed based on maternal serum, cord serum and placental levels at term parturition in women with adequate (n = 15; serum retinol > 20 micrograms/dl) and subadequate (n = 16; serum retinol < or = 20 micrograms/dl) vitamin-A status. There was no difference in retinol and beta-carotene levels in placenta and cord serum between these groups. However, differences in the relation of maternal, placental and cord blood components were observed between women with adequate and subadequate vitamin-A status. In women with subadequate status, circulating fetal retinol levels correlated with placental retinol levels, and maternal serum beta-carotene correlated with placental retinol. Within this group, maternal serum beta carotene and cord serum retinol correlated significantly in women with serum retinol levels lower than 15 micrograms/dl. These results suggest that beta carotene may be a precursor of retinol in placenta and that this conversion may depend on the nutritional status of the mother, being particularly effective in a more depleted state. PMID- 8724651 TI - Extremely low birth weight infants have lower Fc gamma RIII (CD 16) plasma levels and their PMN produce less Fc gamma RIII compared to adults. AB - The purpose of this study was to determine whether decreased Fc gamma RIII expression on the PMN of extremely low birth weight infants (ELBW) is due to decreased receptor synthesis or increased receptor shedding from the PMN surface. 42 ELBW, 12 larger infants and 14 adults were enrolled. Plasma and total cellular Fc gamma RIII were measured by ELISA, and PMN Fc gamma RIII expression was measured by flow cytometry. ELBW PMN plasma membrane expression of Fc gamma RIII as measured by log mean channel fluorescence (5.00 +/- 1.98 vs. 10.68 +/- 1.61, p < 0.050) and plasma Fc gamma RIII levels were both lower (7.5 +/- 6.1 vs. 82.4 +/ 64.8 nM, p < 0.05) than in adult controls. In follow-up studies, 14 ELBW (age = 29 +/- 14 days, range = 14-56 days) increased PMN expression of Fc gamma RIII (p < 0.001) but not plasma Fc gamma RIII. ELBW had lower total PMN-associated Fc gamma RIII than adults (2.3 +/- 0.9 vs. 6.8 +/- 2.2 ng/10(6) PMN, p = 0.006). ELBW's PMN produce less Fc gamma RIII than adults' PMN, and expression of this receptor is developmentally regulated. PMID- 8724652 TI - Doppler assessment of physiological stenosis at the bifurcation of the main pulmonary artery: a cause of functional murmur in neonates. AB - A systolic heart murmur is not infrequently recognized in healthy newborn infants, especially those with a low birth weight. This study aimed at assessing the hemodynamics at the bifurcation of the main pulmonary artery using Doppler echocardiography and to correlate the results with this murmur. The peak velocities of main pulmonary artery (MPAV) and right pulmonary artery (RPAV) were studied in 25 low-birth-weight infants who had recovered from acute stage and presented with a systolic murmur, and the ratio of RPAV/MPAV was calculated. Another compatible 25 healthy low-birth-weight infants without a murmur were enrolled as the control group. The initial MPAV values were 79.8 +/- 20.7 (range 51-152) cm/s and 80.7 +/- 14.2 (range 60-111) cm/s in the heart murmur group and in the control group, respectively (p > 0.05). The initial RPAV values were 193.4 +/- 60.2 (range 118-388) cm/s and 99.8 +/- 15.5 (range 76-132) cm/s in the heart murmur group and in the control group, respectively (p < 0.0001). The initial RPAV/MPAV ratios were 2.46 +/- 0.61 (1.59-3.92) and 1.25 +/- 0.14 (0.94-1.47) in the heart murmur group and in the control group respectively (p < 0.0001). When the murmur disappeared after a period of 2-5 months, no significant differences in the last RPAV and RPAV/MPAV ratios between both groups could be found. The RPAV in the heart murmur group faded significantly to 118.7 +/- 16.9 cm/s, and so did the RPAV/MPAV ratio to 1.24 +/- 0.12 (p < 0.0001). A transient functional murmur recognized in healthy premature infants at about 1 month of age is caused by the pressure gradient that implies a physiological stenosis at the bifurcation of the main pulmonary artery. The RPAV is higher than the MPAV, and the RPAV/MPAV ratio is usually over 1.5. The RPAV became slower, and the RPAV/MPAV ratio fell as the murmur faded with age. PMID- 8724653 TI - Staphylococcus epidermidis sepsis in the intensive care nursery: a characterization of risk associations in infants < 1,000 g. AB - We undertook to determine Staphylococcus epidermidis colonization patterns and risks of sepsis in a cohort of 82 consecutive intensive care nursery admissions (birth weight 1,285 +/- 57 g), with 24 infants weighing < 1,000 g at birth. Colonization was determined by skin and stool cultures collected at three time points. Multiple neonatal variables were classified into three intervals preceding the time of sample collection including the occurrence of S. epidermidis sepsis. 16 infants (20%) developed S. epidermidis sepsis. 81% of these episodes occurred in infants < 1,000 g. Skin colonization was nearly universal at all sampling points. Rectal colonization was 63.6% initially (10 +/- 0.4 days), then declined to 32% by the third sample (37 +/- 0.4 days). Neither prevalence of skin nor rectal colonization influenced the incidence of sepsis significantly. Statistically significant risk associations for sepsis for the entire intensive care nursery population included: low birth weight, gestational age, presence of a central line, and delayed feeding. For infants < 1,000 g the occurrence of sepsis during the second study time period (54% of the episodes) was associated with preceding steroid exposure. During the third study time period, birth weight and delayed attainment of full enteral feeds showed a statistically significant association with sepsis. We conclude that infants < 1,000 g are at an increased risk of S. epidermidis sepsis. Extreme immaturity, steroid therapy, and prolonged hyperalimentation are all significant risk associations. PMID- 8724654 TI - Hypoxic-ischemic injury acutely disrupts microtubule-associated protein 2 immunostaining in neonatal rat brain. AB - We evaluated the influence of an acute hypoxic-ischemic insult on the neuronal dendritic cytoskeletal protein microtubule-associated protein 2 (MAP2) in the immature rat brain. Studies were performed using a well-characterized perinatal rodent stroke model, in which unilateral ischemic forebrain injury was induced in 7-day-old rats by right carotid artery ligation, followed by 3 h exposure to 8% oxygen. Changes in the neuroanatomic distribution of MAP2 in the first 48 h after injury were evaluated by immunocytochemistry with a monoclonal mouse anti-MAP2 antibody. The distribution of MAP2 immunoreactivity was markedly disrupted in the lesioned hippocampus; prominent reductions in MAP2 immunostaining evolved concurrently in lesioned cortex, caudate and thalamus. These data demonstrate that at this developmental stage, MAP2 immunocytochemistry provides a sensitive indicator of the distribution and severity of hypoxic-ischemic neuronal injury. PMID- 8724655 TI - Hemolytic anemia does not increase entry into, nor alter rate of clearance of bilirubin from rat brain. AB - The risk of bilirubin encephalopathy is believed to be increased in hemolytic relative to nonhemolytic jaundice. Young SPRD rats were injected with either acetylphenylhydrazine (APHZ, n = 22) 75 mg/kg or an equivalent volume of the solvent (control, n = 22) intraperitoneally for 3 successive days. One the 4th day, hyperbilirubinemia was induced by a 5-min intravenous infusion of 50 mg/kg bilirubin. After sacrifice (at 10 or 60 min) the brain vessels were flushed in situ with ice-cold saline. Serum bilirubin was 600 +/- 108 mumol/l (mean +/- SD) at 10 min, and 295 +/- 98 mumol/l at 60 min. Hematocrit was significantly reduced in the APHZ rats versus controls (26 +/- 3 vs. 41 +/- 3, p < 0.0001). The concentration of bilirubin in brain was determined by acid chloroform extraction and diazotization. The brain bilirubin values were 5.5 +/- 1.5 versus 5.4 +/- 2.1 mmol/g at 10 min (APHZ vs. control), and 1.3 +/- 0.6 versus 0.8 +/- 0.7 nmol/g at 60 min. The half-life of bilirubin in brain was calculated with an exponential fitting program. The half-life of bilirubin in brain for the two groups was 24.3 +/- 21.9 versus 18.5 +/- 28.3 min. There were no significant differences between the groups in either of these measures. We conclude that in young rats, hemolytic anemia does not increase the acute entry of bilirubin into brain, nor does it affect the clearance of bilirubin from brain. PMID- 8724656 TI - Comparison of natural surfactant and brief liquid ventilation rescue treatment in very immature lambs. Clinical and physiological correlates. AB - We studied the effect of rescue therapy with modified porcine surfactant (Curosurf) or brief perfluorocarbon liquid ventilation on pulmonary gas exchange, mechanics and structure in very immature lambs. Both rescue strategies produced an improvement in arterial oxygenation and were able to support CO2 elimination. Histology showed unevenly inflated lungs in all but tidal-liquid-ventilated lambs. We speculate that due to immature lung architecture and relatively high permeability, previous exposure to gas ventilation impeded full effectiveness of both rescue modalities, perhaps prophylactic surfactant or liquid ventilation could be a better alternative. PMID- 8724657 TI - Inhalation of nitric oxide: effect on cerebral hemodynamics and activity, and antioxidant status in the newborn lamb. AB - Ventilation with nitric oxide (NO) is increasingly being used to treat pulmonary hypertension in the newborn. In the brain, NO has vasoactive properties and is involved in neurotransmission. However, the effect of inhaled NO on the cerebral blood flow (CBF) and on the cerebral activity is not known. Furthermore, there is little information on the influence of this free radical gas on the redox status in pulmonary vessels. We therefore investigated the effect of inhaled NO (2-60 ppm) on CBF, cerebral activity and redox status in blood effluent from the pulmonary circulation in 6 ventilated newborn lambs before and during group B streptococci (GBS)-induced pulmonary hypertension. Blood pressure in the pulmonary artery (P(ap)) and aorta (Pao), carotid artery blood flow (Qcar) to assess changes in CBF, and electrocortical activity were measured. Blood gases, indices of free radical status and methemoglobin were determined in blood samples obtained from the left ventricle. Inhalation of NO, before and during GBS-induced pulmonary hypertension, decreased P(ap) and PCO2 and increased PO2. Multiple linear regression revealed that Qcar was positively related to PCO2, but not to inhaled NO or PO2 before or during GBS conditions. Electrocortical activity and indices of antioxidative capacity and lipid peroxidation did not change significantly. Methemoglobin was not detected. In conclusion, inhalation of NO (up to 60 ppm) lowered P(ap) without directly affecting CBF, electrocortical activity, and redox status in the pulmonary vessels. CBF, however, can indirectly be influenced by NO-mediated changes in PCO2. PMID- 8724658 TI - Host regulation of glial markers in intrastriatal grafts of conditionally immortalized neural stem cell lines. AB - After transplantation into the adult CNS the immortalized neural stem cell lines ST14A and HiB5 differentiate preferentially into glia-like cells. After lesions of the host brain, which activate resting glial populations, the grafted cells responded with up-regulation of glial markers and a change in morphology towards a reactive state. The protein expression followed the same pattern as in the host glial population. The results show that the astrocytes formed by the grafted neural stem cells become functionally integrated in the host brain and that they may take an active part in the reactive gliosis caused by brain damage. PMID- 8724659 TI - Nitric oxide synthase expression in human neuroblastoma cell line induced by cytokines. AB - The human neuroblastoma cell line NB-39-nu expressed mRNA coding for inducible nitric oxide synthase (iNOS) following treatment with a combination of interferon gamma (IFN-gamma) and tumour necrosis factor-alpha (TNF-alpha). The level of iNOS mRNA peaked 24 h after stimulation and had declined by about 25% after 48 h. Trace levels of iNOS mRNA were detected after treatment with IFN-gamma alone, and its mRNA level was synergistically enhanced by simultaneous treatment with TNF alpha. Neither bacterial lipopolysaccharide nor interleukin-1 beta (IL-1 beta) showed synergistic effects as great as that of TNF-alpha on iNOS gene expression. Dexamethasone inhibited the induction of iNOS mRNA by IFN-gamma and TNF-alpha. Induction of iNOS was confirmed by NADPH-diaphorase staining and by immunostaining with human iNOS-specific antibody. PMID- 8724660 TI - GDNF mRNA in Schwann cells and DRG satellite cells after chronic sciatic nerve injury. AB - Glial cell line-derived neurotrophic factor (GDNF) exhibits neurotrophic properties on different types of neurones, including fetal motoneurones and embryonic neurones of sensory ganglia. We demonstrate that chronic injury to the adult rat sciatic nerve induces a rapid up-regulation of GDNF mRNA expression in Schwann cells proximal as well as distal to the injury site, and that expression of this mRNA remains at high levels for at least 5 months after injury. In addition, GDNF mRNA increases and remains high in satellite cells and Schwann cells of the affected L4/L5 DRGs. These findings suggest that GDNF is an important factor in the events that follow upon adult chronic primary sensory neurone injury, and possibly also after adult motoneurone axotomy. PMID- 8724661 TI - Myelin from MAG-deficient mice is a strong inhibitor of neurite outgrowth. AB - Myelin-associated glycoprotein (MAG) has potent neurite outgrowth inhibitory activity in vitro. To assess the importance of MAG in the neurite outgrowth inhibitory activity in CNS myelin, we used an in vitro bioassay to characterize neurite growth on CNS myelin derived from mice carrying a null mutation of the MAG gene. Myelin proteins from MAG-deficient mice inhibited neurite outgrowth to a similar degree to the wild-type CNS myelin. These results suggest that CNS myelin molecules other than MAG exert strong inhibitory effects on the growth of neurites. PMID- 8724662 TI - Na(+)-K(+)-ATPase activity of different regions of rat brain in response to immunization and bone marrow cytokine treatment. AB - The bone marrow cytokine (Bio-Immuno Modulator, BIM or BM-Fr1) has been suggested to correct immunoincompetence by modulating brain Na(+)-K(+)-ATPase. The brain region affected and mechanism of action of BM-Fr1 are unknown, however. Here we report that immunization of immunocompetent rats indirectly inhibited Na(+)-K(+) ATPase activity (59%) in the left cerebral lobe (LC) and irrespective of BM-Fr1 treatment, stimulation of the enzyme was observed in the LC at the peak of immune response. BM-Fr1 treatment, which corrected immunoincompetence in malnourished rats, also modulated a different LC Na(+)-K(+)-ATPase profile to that seen in immunocompetent animals. Immunogen and BM-Fr1 seem to exert their influence in brain via a cytosolic inhibitor protein of Na(+)-K(+)-ATPase. Thus we suggest that (1) BM-Fr1 plays an important role in immune homeostatasis by modulating Na(+)-K(+)-ATPase activity of LC and (2) Na(+)-K(+)-ATPase is not the receptor for either immunogen or BM-Fr1. PMID- 8724663 TI - Long-lasting changes of neuronal activity in the motor cortex of cats. AB - The effect of tetanic intracortical microstimulation (ICMS) of superficial layers of the motor cortex on unit discharges responding to stimulation of the peripheral receptive fields was examined in the motor cortex (area 4y). Single or multiple unit discharges were isolated from 15 recording sites near the stimulating electrodes. Following tetanic ICMS, the response to the same peripheral stimulation showed a long lasting increase at six recording sites, a long lasting decrease at two sites and transient increase or decrease at four sites. In the rest of the sites, tetanic ICMS had no effect. The results demonstrate the existence of use-dependent modification of information processing in the motor cortex and support the hypothesis that the motor cortex participates in learning motor skills. PMID- 8724664 TI - Distribution of histaminergic axonal fibres in the vestibular nuclei of the cat. AB - The histaminergic projection in the cat brain originates from neurones located in the tuberomammillary nucleus. The distribution of histaminergic axonal fibres was investigated immunohistochemically in the vestibular nuclei of the cat using an antibody to histamine. Labelled fibres were sparsely distributed in the whole vestibular nuclei complex and were typically beaded with small, darkly stained swellings. Staining density was low to moderate compared with that of other structures in the cat brain. Histaminergic fibre distribution showed spatial variations, with significantly heavier labelling in the superior and medial vestibular nuclei than in the lateral and inferior nuclei. Histamine could play a neuromodulatory role in the processing of sensory afferent information in the vestibular nuclei and in the control of vestibular functions. PMID- 8724665 TI - Cerebral activation covaries with movement rate. AB - An important aspect in brain activation studies is the relationship between neuronal activity and measurable indices of function. We applied functional magnetic resonance imaging (fMRI) to investigate blood flow-related MR signal changes in response to different rates of repetitive movements of the index finger. The contralateral precentral gyrus and the posterior frontomesial cortex revealed a significant increase in MR signal over baseline for 1, 2 and 3 Hz finger movements, with a linear effect of rate in the precentral gyrus. Increased firing of neuronal aggregates or recruitment of additional neuronal units within the primary motor cortex necessary for increased output to target neurons and maintaining posture of nearby distal and proximal joints may contribute to the activation pattern. PMID- 8724666 TI - Neuronal responses in the motion pathway of the macaque monkey to natural optic flow stimuli. AB - Neurones in higher visual motion areas in the superior temporal sulcus (STS) of the macaque monkey respond to abstract random dot optic flow stimuli. Higher motion areas may not only represent, but in a next computational stage also analyse the flow field to determine, for instance, the direction of heading for navigation purposes. Real world visual scenes differ in several aspects from these abstract optic flow stimuli. We tested the neuronal response to naturalistic optic flow stimuli which simulated egomotion in different virtual environments and contained different numbers of visual cues. Neuronal activity depended mainly on the position of the focus of expansion rather than on other visual cues. This finding supports the hypothesis that higher motion areas within the STS analyse optic flow in natural scenes and can thus signal the direction of heading. PMID- 8724667 TI - NT-3 and/or BDNF therapy prevents loss of auditory neurons following loss of hair cells. AB - Destruction of auditory hair cells results in a subsequent loss of auditory neurons. In situ hybridization and neuronal cell culture studies as well as analyses of the inner ears of neurotrophin and neurotrophin receptor gene knockout mice have shown that NT-3 and BDNF mediate both the development and survival of auditory neurons. In this study guinea pigs were exposed to the ototoxic combination of an aminoglycoside antibiotic and a loop diuretic and then received 8 weeks of intracochlear infusion of either NT-3, BDNF or NT-3 + BDNF to determine whether site-specific application of these neurotrophins could prevent the loss of auditory neurons that follows a loss of auditory hair cells. Infusion of either NT-3 or NT-3 + BDNF into the scala tympani resulted in a > 90% survival of auditory neurons while BDNF infusion yielded a 78% survival rate, compared with a 14-24% neuronal survival rate in untreated ototoxin-exposed cochleae. These results show that loss of auditory neurons that occurs subsequent to a loss of auditory hair cells can be prevented by in vivo neurotrophin therapy with either NT-3 or BDNF. PMID- 8724668 TI - Peripheral administration of NMDA, AMPA or KA results in pain behaviors in rats. AB - The present study investigated the role of N-methyl-D-aspartate (NMDA) and non NMDA glutamate receptor subtypes in peripheral pain transmission. Activation of NMDA, alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) and kainate acid (KA) receptors in glabrous skin of the rat hindpaw resulted in mechanical allodynia and mechanical hyperalgesia. These agonist-induced pain behaviors were attenuated following peripheral injection of appropriate antagonists (MK-801 and CNQX). Thus, activation of NMDA, AMPA or KA receptors at the level of the peripheral nerve terminal can produce nociceptive behavior. These data suggest that topical application of glutamate receptor antagonists may be useful in treating pain disorders. Since all three receptor subtypes are involved in peripheral pain transmission, however, it will be necessary to antagonize multiple glutamate receptor subtypes to achieve effective pain relief. PMID- 8724669 TI - Comparative autoradiographic distribution of neuropeptide Y Y1 receptors visualized with the Y1 receptor agonist [125I][Leu31,Pro34]PYY and the non peptide antagonist [3H]BIBP3226. AB - BIBP3226, a non-peptide neuropeptide Y Y1 receptor antagonist, was used in its tritiated form to investigate the autoradiographic distribution of its binding sites in the rat brain. [125I][Leu31,Pro34]pPYY a recently developed Y1 agonist radioligand was used for comparison. High levels of specific binding for both radioligands were seen in various areas of the rat brain, including superficial cortical laminae, anterior olfactory nucleus, claustrum, geniculate nuclei and medial mamilliary nucleus. However, a few regions, such as the external plexiform layer of the olfactory bulb, lateral septum nucleus tractus, solitarius and area postrema were only enriched with specific [125I][Leu31,Pro34]pPYY binding sites. These data suggest the possible heterogeneity of [125I][Leu31,Pro34]pPYY binding sites in the rat brain, a sub-population of which apparently fails to be recognized by the newly developed Y1 receptor antagonist, BIBP3226. PMID- 8724670 TI - Brain potentials as objective indexes of auditory pitch extraction from harmonics. AB - Complex tones such as vowels expressed by the human voice vary both in fundamental pitch (harmonic periodicity) and spectrum shape (vowel type). Three different pitches and spectra were combined into nine auditory stimuli. Latencies of cerebral evoked potential components decreased significantly as pitch increased, whereas they remained constant with spectrum shape. Because the fundamental (direct clue about pitch value) was purposely omitted from stimuli, these results reflect feature extraction processes. They also demonstrate that brain potentials can be used as objective indexes of perceived pitch. PMID- 8724671 TI - Dopamine deficits in the brain: the neurochemical basis of parkinsonian symptoms in AIDS. AB - We sought to determine whether the motor dysfunctions and neuroleptic sensitivity that can occur in patients with AIDS relates to a deficit of striatal dopamine innervation similar to that of Parkinson's disease. For this purpose we measured concentrations of dopamine and its major metabolite homovanillic acid (HVA) in caudate nucleus tissue taken post-mortem from patients with AIDS and from appropriate age-matched control subjects. Dopamine and HVA concentrations were both significantly reduced in the AIDS group, with 20 of 34 patients exhibiting dopamine concentrations below the control range. This finding is consistent with a loss of nigrostriatal dopaminergic neurones, and may underlie the motor dysfunction and neuroleptic sensitivity that can occur in AIDS patients, indicating the potential value of antiparkinsonian therapy in such patients. PMID- 8724672 TI - Effects of neurotrophins on early auditory neurones in cell culture. AB - During the first week of postnatal development, the innervation of the organ of Corti changes from an immature to an adult pattern. Dissociated cell cultures of early postnatal spiral ganglia were used to investigate the effects of nerve growth factor (NGF), brain derived neurotrophic factor (BDNF) and neurotrophin-3 (NT-3) on maturing auditory neurones. BDNF was the most potent stimulator of neuritogenesis, NT-3 provided the strongest support for neuronal survival, while NGF supported limited neuritogenesis, and only at pharmacological levels. These findings suggest that both BDNF and NT-3 participate in the postnatal maturation of cochlear innervation and that NGF is most probably not involved in this process. PMID- 8724673 TI - Pupil dilatation assay by tropicamide is modulated by apolipoprotein E epsilon 4 allele dosage in Alzheimer's disease. AB - The mydriatic response to dilute tropicamide was studied in 25 Japanese patients with Alzheimer's disease (AD), 13 patients with non-AD neurological diseases (non AD) and 11 normal elderly subjects (control). Although the changes in resting pupil diameter and area over baseline were significantly greater (p < 0.05) in AD patients than in non-AD patients and controls, there was considerable overlap between the three groups. The change in resting pupil area was significantly greater (p < 0.05) in AD patients homozygous for ApoE epsilon 4 than in AD patients heterozygous for or without this allele. Despite a limited sample size in the present study, our results indicate that the pupil dilation assay by tropicamide is not an effective diagnostic tool for AD, and it may be modulated by different gene dosage of ApoE epsilon 4. PMID- 8724674 TI - Glutathione depletion potentiates MPTP and MPP+ toxicity in nigral dopaminergic neurones. AB - Glutathione levels are decreased in the substantia nigra of patients with Parkinson's disease. We studied whether glutathione depletion contributes to dopaminergic cell death using a specific inhibitor of glutathione biosynthesis, L buthionine sulfoximine (BSO). We found no significant reduction of tyrosine hydroxylase-positive cells in the substantia nigra pars compacta (SNpc) when BSO was administered systemically to preweanling mice or locally to the SNpc of adult rats. However, the combination of BSO with MPTP (1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine) in preweanling mice and the combination of nigral injections of BSO with intrastriatal injections of MPP+ (1-methyl-4-phenylpyridinium), the active metabolite of MPTP in adult rats, potentiated the toxic effects of MPTP and MPP+ on nigral neurones. Our data show that glutathione depletion can result in cell death if the nigrostriatal system is metabolically compromised. PMID- 8724675 TI - Accumulation of NO synthase (type-I) at the neuromuscular junctions in adult mice. AB - Recent data have shown that NOS-I is localized almost exclusively to the sarcolemma of fast-twitch fibers, where it probably interacts with the dystrophin glycoprotein complex. The concentration of dystrophin-related protein at the neuromuscular junctions and the possible involvement of NO in synaptic suppression led us to investigate the presence of NOS-I at the adult motor endplates. Our data clearly show that NOS-I protein, detected by immunohistochemistry accumulates at the adult endplates. Furthermore, the absence of NOS-I protein at the denervated neuromuscular junctions suggest a neural origin of this enzyme. The putative roles of NO at the endplate are discussed. PMID- 8724676 TI - Adrenocortical activation by herpes virus: involvement of IL-1 beta and central noradrenergic system. AB - We investigated the involvement of brain cytokines and central monoamines in mediating the effect of the neurotropic herpes simplex virus-1 (HSV-1) on adrenocortical activity in rats. Corneal inoculation with a neurovirulent HSV-1 strain, but not with an avirulent strain, induced interleukin-1 beta (IL-1 beta) gene expression mainly in the pons and hypothalamus, and caused an elevation in serum corticosterone levels. Infectious virus was isolated in low titres only from the trigeminal ganglia and pons. Viral DNA was detected by PCR in these tissues and in other brain regions. Virus-induced adrenocortical activation was abolished in rats in which hypothalamic norepinephrine (NE) was depleted by 6 hydroxydopamine. Depletion of hypothalamic serotonin by 5,7-dihydroxytryptamine did not prevent adrenocortical activation. These results suggest that central IL 1 and NE are involved in HSV-1 induced adrenocortical activation. PMID- 8724677 TI - Auditory hallucinations inhibit exogenous activation of auditory association cortex. AB - Percepts unaccompanied by a veridical stimulus, such as hallucinations, provide an opportunity for mapping the neural correlates of conscious perception. Functional magnetic resonance imaging (fMRI) can reveal localized changes in blood oxygenation in response to actual as well as imagined sensory stimulation. The safe repeatability of fMRI enabled us to study a patient with schizophrenia while he was experiencing auditory hallucinations and when hallucination-free (with supporting data from a second case). Cortical activation was measured in response to periodic exogenous auditory and visual stimulations using time series regression analysis. Functional brain images were obtained in each hallucination condition both while the patient was on and off antipsychotic drugs. The response of the temporal cortex to exogenous auditory stimulation (speech) was markedly reduced when the patient was experiencing hallucinating voices addressing him, regardless of medication. Visual cortical activation (to flashing lights) remained normal over four scans. From the results of this study and previous work on visual hallucinations we conclude that hallucinations coincide with maximal activation of the sensory and association cortex, specific to the modality of the experience. PMID- 8724678 TI - Stochastic particle formulation of the vesicle hypothesis. Relevance to short term phenomena. AB - Based on the vesicle hypothesis, the modes of elementary quantum-vesicle interactions have been formulated in terms of probabilities of induced and spontaneous interstate quanta transitions and generalized within the framework of the previously developed theory of the double barrier synapse. Among the three allowed states for a quantum, the transition state is a novel formulation for the fraction of immediately available quanta governed by both vesicle and presynaptic membranes. The parameters of the model were determined by fitting solutions to the experimental curves representing effects of single pulse and short train activation on transmitter release at the frog neuromuscular junction. Qualitative differences in particle physics of transmitter release found under low quantal outputs on one hand and normal transmission on the other allowed the formulation of the uncertainty hypothesis and the quantum condition of synaptic homeostasis. PMID- 8724679 TI - The agonistic action of pentobarbital on GABAA beta-subunit homomeric receptors. AB - Murine gamma-aminobutyric acid type A (GABAA) receptor beta 1, beta 2, and beta 3 subunits were expressed in Xenopus oocytes and studied using the two electrode voltage clamp technique. Although all three beta-subunits were unresponsive to GABA when expressed as homomers, the intravenous general anaesthetics pentobarbital, etomidate and propofol induced currents in beta 2 and beta 3 homomers. The pentobarbital-induced currents in beta 3 homomers showed a dose dependence with an ED50 of 89 +/- 8.9 microM and a Hill coefficient of 0.94 +/- 0.08. Zinc (50 microM) blocked (61.1 +/- 5.6% of control) and 200 microM lanthanum potentiated (139 +/- 8.6% of control) the pentobarbital-induced current. This current was also blocked by picrotoxin but was insensitive to the GABAA receptor antagonist bicuculline. These observations indicate that the full expression of the agonistic action of GABA requires the presence of an alpha subunit, in contrast to the agonistic action of intravenous general anesthetics, where the presence of a beta2 or beta 3-subunit is sufficient. The difference in the agonistic action of intravenous anaesthetics among these highly homologous beta-subunits suggests that the beta-subunit homomeric receptors may be useful to further define the molecular sites of action of intravenous general anaesthetics and other functional domains on GABAA receptors. PMID- 8724680 TI - Persistence of CA1 hippocampal LTP after selective cholinergic denervation. AB - The possible role of endogenous cholinergic innervation in hippocampal plasticity is controversial. We studied the role of acetylcholine (ACh) in short- and long term potentiation (STP and LTP), using the cholinergic neurotoxin 192 IgG saporin. It was still possible to induce STP the LTP in the CA1 field following complete and selective cholinergic denervation of the hippocampus. This study therefore demonstrates that integrity of the endogenous cholinergic system is not necessary for the induction or maintenance of LTP in the CA1 field of the hippocampus. The consequences in terms of relationship between hippocampal cholinergic system, LTP and memory are discussed. PMID- 8724681 TI - Distribution of a rat galanin receptor mRNA in rat brain. AB - In situ hybridization histochemistry has been employed to determine the distribution of the mRNA encoding a recently cloned rat galanin receptor (rGalR1). The galanin receptor mRNA has been found to be discretely localized in rat brain. The most intense hybridization signals were found over neurons in the nucleus of the lateral olfactory tract, in the ventral posterior hippocampus, and in the lateral external subdivision of the parabrachial nucleus. A number of other brain regions also contain significant hybridization signals, including the hypothalamus, brain stem and spinal cord. The localization of rGalR1 mRNA indicates that this receptor may play a role in the varied functions ascribed to GAL, among them feeding, cognition and modulation of sensory information. PMID- 8724682 TI - NADPH-diaphorase-containing enteric neurones survive for a year in the adult rat striatum. AB - In previous studies, we have demonstrated that enteric ganglia can survive when transplanted into the striatum. However, if such grafts are to be effective in clinically significant situations, it is necessary for them to survive in the brain for long periods. In this study, we have examined the corpus striatum of host rats one year after transplantation of pieces of myenteric plexus taken from young Fischer rats. NADPH-diaphorase-containing enteric neurones had survived within the CNS environment one year after grafting, and had extended axons into the surrounding striatum. PMID- 8724683 TI - Localizing the lexicon for reading aloud:replication of a PET study using fMRI. AB - Functional magnetic resonance imaging (fMRI) was used to investigate the neural basis of written word recognition in two normal subjects. With a 1.5T scanner and temporal surface coil, T2 gradient echo images were obtained while subjects read words aloud. As a control condition, subjects visualized false font strings and said the word "range' each time such a string appeared. These two conditions were presented in an oscillatory pattern, alternating 30 s of each condition for a total of 4 min. Comparison of the two conditions using cross-correlation demonstrated strong activation in both subjects in the left posterior superior temporal gyrus, near the site predicted for the visual input lexicon by Dejerine and recently demonstrated by positron emission tomography. PMID- 8724684 TI - Structural interactions between NOS-positive neurons and blood vessels in the hippocampus. AB - The anatomical relationship between nitric oxide synthase (NOS)-positive neurons and blood vessels was examined in the hippocampus of the rat. NADPH-diaphorase histochemistry was used to identify NOS-positive neurons by light-microscopy. A close association of somatic, dendritic and axonal processes of NOS-positive neurons with cerebral blood vessels was observed. These findings suggest the possibility of neurovascular signaling by local NOS-containing neurons, through direct vascular innervation by terminals generating nitric oxide, and paracrine signaling from closely apposed somatic and dendritic neuronal elements. PMID- 8724685 TI - The evolution of customer feedback in health care. PMID- 8724686 TI - Learning from patients: a discharge planning improvement project. AB - BACKGROUND: In 1991 Beth Israel Hospital (Boston) joined nine other hospitals in using the Picker/Commonwealth survey instrument to tap patients perceptions of their hospitalization experience. Beth Israel focused on one of the nine dimensions of the instrument-continuity and transition (discharge planning). FOUR WORK TEAMS: In 1992 four multidisciplinary work teams were formed-for cardiac surgical patients, stroke patients, patients on a medical unit, and patients on a medical and surgical unit. Each team conducted a patient/family discussion group, during which recently discharged patients and their families were asked about their preparation for discharge and asked for input on how to improve the process. INTERVENTIONS: Each work team developed interventions on the basis of information specific to their patients. The cardiac work team, for example, developed interdisciplinary practice guidelines for patient care management for the entire postoperative period; the guidelines include a patient education component on what patients and families can expect during hospitalization. OUTCOMES: Clinicians practice differently, inviting more patient feedback and other involvement in care, as a results of their involvement in the project. On the first annual patient survey, administered in 1994, only 6% of 1,179 randomly selected patients (versus 20% of the 100 patients first surveyed in 1993) indicated that they did not receive the information they needed to help themselves recover. CURRENT PROGRESS AND FUTURE DIRECTIONS: A standardized teaching packet containing material developed during the discharge planning improvement project is now distributed. In May 1995 the nursing department launched a patient and family learning center to better meet the health education needs of patients. PMID- 8724687 TI - Using qualitative and quantitative patient satisfaction data to improve the quality of cardiac care. AB - BACKGROUND: In early 1993 leaders within the Hitchcock Clinic and cardiac services section at Dartmouth-Hitchcock Medical Center (Lebanon, NH) formed the Cardiac Services Improvement Group (CSIG) as a pilot program for patient-centered quality improvement (QI) at the sectional level. CSIG PROGRAM: For open heart surgery (OHS) and percutaneous transluminal coronary angioplasty (PTCA), a flowchart was constructed of sequential patient experiences. Content analysis of focus group discussions resulted in six key patient-defined quality characteristics: comfort, caring, certainty, convenience, communication, and cost. Linking of patient comments to points on the patient experience flowchart made it possible to determine where particular quality characteristics were most relevant. A patient satisfaction survey with questions that were specific to a patient experience and to a quality characteristic was mailed to 100 consecutively discharged OHS and PTCA patients; 35 of the 50 patients in each group responded. EVALUATING SUCCESS: Analysis of the survey results led to the formation of two QI teams. One team began work on development of a critical pathway for discharge preparation and identified marker questions to track and monitor pathway success in subsequent surveys. Another team began the development of protocols for more effective pain management during and after the PTCA procedure. A repeat patient satisfaction survey that took place from November 1994 to March 1995 suggested a global improvement in patient satisfaction for all patient experiences and all key quality characteristics. CONCLUSION: The CSIG pilot program of patients-based quality measurement and management at the sectional level has been successful in fostering QI team formation and has been associated with a positive deflection in patient-based quality measures. Additional sections of the medical center have initiated similar projects, beginning with process definition and focus groups. PMID- 8724688 TI - Payer-hospital collaboration to improve patient satisfaction with hospital discharge. AB - BACKGROUND: Brigham and Women's Hospital, in Boston, and its major health maintenance organization (HMO), Harvard Community Health Plan, collected data in spring 1994 which revealed that patients were less satisfied with hospital discharge planning than with other elements of care. PROBLEM IDENTIFICATION PROCESS: An interdisciplinary team, formed in November 1994 and composed of eight members from the hospital and HMO, used data from the hospital's Patient Satisfaction Survey, flowcharting, and phone interviews with patients to identify discharge planning-related problems. For example, follow-up contact with patients after discharge was erratic and no clear signal of the successful "hand off" of care from the hospital team to the community team existed. IMPROVEMENT CYCLE 1: Eighty-three percent of the payer's patients that received the improvement strategy developed by the interdisciplinary team-a concierge service-rated discharge planning as excellent or very good, compared to 63% of control patients. IMPROVEMENT CYCLE 2: PLAN SOLUTIONS/STRATEGIES FOR IMPROVING HOSPITAL DISCHARGE PLANNING: The results of the team's Cycle 1 improvement provided information for the team to use in designing a second cycle of incremental improvement activity. For example, to address the lack of clarity about who was responsible for making decisions about discharge and follow-up care, the attending physician was designated the transition-of-care coordinator. Once all the improvements were implemented, Cycle 2 patients who received the intervention rated satisfaction with discharge higher (83% versus 73%) than the control group. CONCLUSIONS: Implications of hospitalwide implementation of discharge planning related services attempted on one unit are being considered. PMID- 8724689 TI - Linking customer judgments with process measures to improve access to ambulatory care. AB - BACKGROUND: The true value of patients satisfaction emerges when findings are linked with process data to direct and measure the results of continuous improvement efforts. In late 1991 Henry Ford Medical Group, with its ambulatory care facilities, began surveying internal and external customers regarding access to care. IDENTIFYING THE ACCESS ISSUES: Creating a system diagram and defining the core steps in the primary care visit process proved instrumental in identifying access issues. Focus group research and interviews with internal and external customers led to a list of "current reality" statements, which proved useful in defining process barriers and identifying potential improvement strategies. In June 1993, at the first of a series of retreats on improving access to primary care, four physician-led subgroups were formed to address key access strategies, one of which focused on the availability of telephone triage and nursing-advice services. PLAN OF ACTION: Key strategies for the triage/advice subgroup focused on upgrading phone systems, identifying dedicated space for triage/advice activity, and developing consistent advice guidelines. USING DATA TO PLAN PROCESS IMPROVEMENTS: Once upgrading had occurred in several sites, local quality improvement teams were formed to address specific phone access issues. Data suggested that clinics could use call volume to construct appointment schedules for adequate same-day access. Tracking and trending the call volume data could permit facilities to adjust provider schedules to meet daily and seasonal fluctuations in demand. CONCLUSIONS: The combination of key process and patient survey data have allowed for identification of system deficiencies and monitoring of improvement efforts. PMID- 8724690 TI - Benchmarking patient satisfaction at academic health centers. AB - BACKGROUND: In 1991 the University HealthSystem Consortium (UHC), an alliance of 70 academic health centers, began its patient satisfaction benchmarking project. The survey, adapted from the Picker Institute survey, was pilot tested in 1992 and has been in use since 1993. Each year the project's steering committee refines the survey on the basis of member needs and survey item performance. KEY FINDINGS: Findings have shown that the survey can document the effects of specific quality improvement efforts, that patients from different medical services report different levels of satisfaction with their care, and that physician and nursing care have had the greatest impact on overall satisfaction. USING THE RESULTS: Each participating organization receives concise narrative reports of the survey results, with priorities for improvement efforts clearly highlighted. A five-to six-page Executive Summary provides the organization's executive team with a quick overview of the results, as well as a summary of the areas where quality improvements are most needed. A longer Managers' Report provides a more detailed analysis of the findings for quality managers and department heads. Sections for each major area of care can be copied and distributed as "stand alone" reports to the appropriate decision makers. For example, the section on nursing care can be distributed to the chief nursing officer and nurse managers. For each key aspect of the patient's experience, best practices for maintaining patient satisfaction are identified from across the hospitals and compiled into a catalogue. LESSONS LEARNED: The UHC patient satisfaction benchmarking program has created ongoing communication among the participating hospitals, whose staff members have been willing to share problems encountered and possible solutions. PMID- 8724691 TI - Designing surveys for maximal satisfaction:an interview with Allyson Ross Davies. Interview by Maggie Kennedy. AB - The usefulness of patient feedback data depends in large part on the relevance and effectiveness of the feedback mechanism used-whether a satisfaction survey, focus group, or one-on-one interview. In the following interview conducted in March 1996, Allyson Ross Davies, PhD, MPH, discusses an approach for designing a patient satisfaction survey that will help organizations gather useful information for improvement activities and other organizational activities. PMID- 8724692 TI - Vav: function and regulation in hematopoietic cell signaling. AB - Vav, a 95 kDa proto-oncogene product expressed specifically in hematopoietic cells, was originally isolated as a transforming human oncogene. Vav contains an array of functional domains that are involved in interactions with other proteins and, possibly, with lipids. These include, among others, a putative guanine nucleotide exchange domain, a cysteine-rich region similar to the phorbol ester/diacylglycerol-binding domain of protein kinase C, a pleckstrin-homology domain, and Src-homology 2 and 3 (SH2 and SH3, respectively) domains. The presence of these domains, the transforming activity of the vav oncogene, and the rapid increase in tyrosine phosphorylation of Vav induced by triggering of diverse receptors indicate that it plays an important role in hematopoietic cell signaling pathways. Such a role is supported by recent studies using "knockout" mice and transiently transfected T cells, in which Vav deletion or overexpression, respectively, had marked effects on lymphocyte development or activation. The presence of a putative guanine nucleotide exchange domain, the prototype of which is found in the dbl oncogene product, implies that Vav functions as a guanine nucleotide exchange factor (GEF) for one (or more) members of the Ras-like family of small GTP-binding proteins. In support of such a role, Vav preparations were found in some (but not other) studies to mediate in vitro specific GEF activity for Ras. Additional studies are required to identify the physiological regulators and targets of Vav, and its exact role in hematopoietic cell development and signaling. PMID- 8724693 TI - Hematopoietic stem cells in the mouse embryonic yolk sac. AB - The yolk sac is the first site of hematopoiesis during mammalian development. The yolk sac is also the first site of blood vessel development. Development of the blood islands in the yolk sac is an integrated process in which these two developmental events, hematopoiesis and vasculogenesis, proceed in concert. This review focuses on mouse yolk sac hematopoietic stem cells (YS-HSC), describing their differentiation in vitro and in vivo. YS-HSC go through a progressive series of changes prior to the initiation of lineage-specific differentiation. Experiments tracing their origins from postulated hemangioblasts, and the subsequent interaction between these stem cells and yolk sac endothelial cells are described. Differences between the extraembryonic YS-HSC and HSC found later within the embryo, perinatally or in adults, are described. YS-HSC have greater reproductive capability than HSC obtained from fetal liver, umbilical cord blood or adult bone marrow; they do not yet express major histocompatibility complex associated antigens and they are able to reconstitute adult immunocompromised animals even when introduced in small numbers (< 100 cells/mouse). With recent results demonstrating the feasibility of expanding YS-HSC in vitro as well as of introducing new genes into these cells by transfection, the YS-HSC shows promise both as a means of achieving long-term restitution of hematopoiesis across histocompatibility barriers and as a self-renewing vehicle for gene transfer. PMID- 8724694 TI - The role of HOX homeobox genes in normal and leukemic hematopoiesis. AB - A sizable amount of new data points to a role for the HOX family of homeobox genes in hematopoiesis. Recent studies have demonstrated that HOXA and HOXB genes are expressed in human CD34+ cells, and are downregulated as cells leave the CD34+ compartment. In addition, expression of certain genes, including HOXB3 and HOXB4, is largely restricted to the long-term culture-initiating cell enriched pool, containing the putative stem cell population. Studies have also shown that HOX genes appear to be important for normal T lymphocyte and activated natural killer cell function. Overexpression of Hox-b4 in transplanted murine marrow cell results in a dramatic expansion of stem cells, while maintaining normal peripheral blood counts. In contrast, overexpression of Hox-a10 resulted in expansion of progenitor pools, accompanied by unique changes in the differentiation patterns of committed progenitors. Overexpression of Hox-a10 or Hox-b8 led to the development of myeloid leukemias, while animals transfected with marrow cells overexpressing Hox-b4 do not appear to develop malignancies. Blockade of HOX gene function using antisense oligonucleotides has revealed that several HOX genes appear to influence either myeloid or erythroid colony formation. Mice homozygous for a targeted disruption of the HOX-a9 gene show reduced numbers of granulocytes and lymphocytes, smaller spleens and thymuses, and reduced numbers of committed progenitors. These studies demonstrate that HOX homeobox genes play a role in both the early stem cell function as well as in later stages of hematopoietic differentiation, and that perturbations of HOX gene expression can be leukemogenic. PMID- 8724695 TI - Granulocyte-macrophage colony-stimulating factor overrides the immunosuppressive function of corticosteroids on rat pulmonary dendritic cells. AB - Pulmonary dendritic cells (DC) are present in extremely small numbers, but they are the most potent antigen-presenting cells in the lungs. Pure populations of DC can be isolated from the lung following collagen digestion, Percoll gradient centrifugation, removal of phagocytic cells and flow cytometric sorting for cells which exhibit high levels of surface major histocompatibility complex (MHC) class II molecules. Exogenous GM-CSF enhances this immunostimulatory capacity of the pulmonary DC. Soluble factors produced by type II airway epithelial cells and interstitial macrophages also enhance the immunostimulating capacity of pulmonary DC while alveolar macrophages suppress it. Thus, the function of DC may be regulated by locally produced cytokines. Corticosteroids are widely used as immunosuppressive agents in pharmacotherapy. While these agents are known to inhibit T cell proliferation and macrophage activation, their effects on DC are not known. We found that dexamethasone (Dex) pretreatment resulted in about a 50% reduction in the immunostimulatory capacity of rat pulmonary DC. This was associated with downregulation of MHC class II (Ia) expression. Dex-induced suppression of DC function could be restored with GM-CSF. We conclude that corticosteroids downregulate antigen-presenting capacity by direct suppression of pulmonary DC. This immunosuppressive effect of corticosteroids on DC may, however, be abrogated by exogenous GM-CSF. Corticosteroids and GM-CSF are therapeutic agents with potent direct immunomodulating effects on DC. PMID- 8724696 TI - Minimal residual disease post-bone marrow transplantation for hemato-oncological diseases. AB - The detection of minimal residual disease (MRD), which is important in cancer treatment, gained special significance in bone marrow transplantation (BMT-) due to the possibility not just to detect but recently also to prevent, treat and reinduce remission in patients that relapsed post-BMT by immunotherapy. The various modern techniques of MRD detection are described including cytogenetics, analysis of restriction fragment length polymorphism, variable number of tandem repeats by Southern Blot or polymerase chain reaction (PCR), microsatellite sequences, PCR amplification products of the Y chromosome or the Amelogenin gene, quantitative PCR and fluorescence in situ hybridization. The role of MRD detection in refinement of indications for BMT, autografting, prediction of relapse, adoptive immunotherapy, mixed chimerism in nonmalignant diseases and in solid organ transplantation is discussed. PMID- 8724697 TI - A review of the costs, cost-effectiveness and third-party charges of bone marrow transplantation. AB - In an era of health care reform, costs, cost-effectiveness and charges for health care are taking on a larger role in the decision to adopt a new therapy. Hospitals, health maintenance organizations, third-party payers, doctors and patients all have an interest in the economic factors of health care; however, not all new therapies or medications have been analyzed. Bone marrow transplantation is one of the most expensive cancer treatments, costing an average of +193,000 per patient; therefore, many economic studies have focused on the costs of the therapy. However, it is important to note that these findings are not necessarily applicable to other diseases. Cost data are not static and even small changes in protocol can make a large difference in costs. Also, cost data from outside the United States is not always applicable to the United States system due to differences such as socialized medicine, workers' wages and pharmaceutical costs. Many economic analyses focus on the hospitals' charges and extrapolate cost-effectiveness from these figures, yet the amount hospitals charge for services and products is not always relative to the costs. Therefore, third-party payer costs are also important in analyzing the cost-effectiveness of a procedure. This article will review five signature papers which illustrate important issues to consider when trying to determine the costs and cost effectiveness of bone marrow transplants. PMID- 8724698 TI - Flow cytometric analysis of megakaryocyte-associated antigens on CD34 cells and their progeny in liquid culture. AB - Three-color flow cytometry was used to analyze the coexpression of surface antigens on megakaryocytes (MKs) developing in liquid cultures of enriched CD34+ cells purified from cord blood. Cells were cultured in serum-replete medium supplemented with interleukin 3 (IL-3), stem cell factor and IL-6. During two weeks of culture, total cells increased 76 +/- 36-fold. CD34+ cells maximally expanded between days 2 and 4, and then gradually decreased to their original input numbers by day 14. As CD34+ cells declined, MKs, defined as glycoprotein (GP) IIbIIIa+ cells, steadily increased in culture 20.9 +/- 18.3-fold. Megakaryopoiesis was further defined by monitoring the expression of GPs IIb, IIIa, Ib, IbIX, and IIIb and c-kit antigen. Increased expression of GPs IIbIIIa and IIb occurred earliest in culture, followed by IIIa and Ib, and then IbIX. Expression of IIIb, also found on monocytes, did not parallel that of the other antigens except when coexpressed on IIbIIIa+ cells. c-kit expression paralleled that of CD34 until the second week of culture when expression was high on nonMKs. Each of these antigens was coexpressed on CD34+ cells and identified a subset of late MK progenitors that increased steadily in culture. Triplelabeled cells expressing CD34, IIbIIIa and a third MK-related antigen were seen at all times. Polyploid MKs of up to 32N were observed during the second week of culture. Multiparametric flow cytometry proved to be a rapid, sensitive and specific method for quantitating the changes in antigen expression of differentiating MKs. PMID- 8724699 TI - Thrombopoietin suppresses apoptosis and behaves as a survival factor for the human growth factor-dependent cell line, M07e. AB - Thrombopoietin (TPO) has been demonstrated to have proliferative effects on hematopoietic progenitor cells and maturational effects on more committed populations which express a megakaryocyte lineage-specific phenotype. M07e is a GM-CSF or interleukin 3 (IL-3)-dependent human leukemic cell line having surface markers characteristic of both myeloid progenitors and megakaryocytes. The effects of TPO on the proliferation and survival of M07e cells were investigated. Following an 18-h factor starvation period to remove residual growth factor signals and phase the cells in G0/G1, TPO provides a weak proliferative signal to M07e compared to IL-3 or GM-CSF treatment under the same conditions. However, TPO synergizes with both GM-CSF and IL-3, and to a greater extent with steel factor, a competence factor for M07e, in the induction of cellular proliferation. TPO sustains cellular integrity of M07e during prolonged (18 days) growth factor withdrawal and also protects M07e cells in serum-free conditions. In addition, preincubation of M07e for 72 h in TPO maintains its survival for subsequent cytokine-induced proliferation, while control media do not. TPO suppresses growth factor withdrawal-induced apoptosis as evaluated by flow cytometric detection of both in situ terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling and cellular DNA content via propidium iodide staining. These results suggest a role for TPO as a survival factor for M07e cells. PMID- 8724700 TI - Comparative effects of insulin-like growth factor II (IGF-II) and IGF-II mutants specific for IGF-II/CIM6-P or IGF-I receptors on in vitro hematopoiesis. AB - This report presents the results of studies investigating the effect of insulin like growth factor II (IGF-II) on the proliferation and differentiation of CD34+ bone marrow cells in serum-substituted liquid cultures. Bone marrow cells were enriched for CD34+ cells and then placed in liquid cultures supplemented with either interleukin 3 (IL-3) or IL-3 and c-kit ligand with and without the addition of IGF-II. When CD34+ cells were incubated with IL-3, cellularity increased throughout four weeks of culture. Cellularity was twofold greater when cultures also contained IGF-II. IGF-II also promoted an increase in cellularity in cultures with IL-3 and c-kit ligand. In combination with IL-3 or IL-3 and c kit ligand, IGF-II promoted an earlier differentiation of granulocytes, as well as an increase in the number of megakaryocyte lineage cells. There were approximately two-fold more colony-forming units for granulocytes and macrophages (CFU-GM) and burst-forming units for erythroid cells (BFU-E) in cultures containing both IL-3 and IGF-II than in cultures with IL-3 alone. These results demonstrate that in cytokine-supplemented media, physiological concentrations of IGF-II augmented both the proliferation and differentiation of CD34+ bone marrow cells while maintaining a greater number of progenitor cells. To identify the receptors through which IGF-II enhances in vitro hematopoiesis, IGF-II was substituted with one of the mutant forms of IGF-II that selectively interacts with either IGF-II/CIM6-P receptors or with IGF-I and insulin receptors. The results with the mutant forms of IGF-II demonstrate that IGF-II augments in vitro hematopoiesis primarily through its interaction with IGF-I and possibly insulin receptors, rather than IGF-II/CIM6-P receptors. PMID- 8724701 TI - 2',2'-Difluorodeoxycytidine (gemcitabine) induces apoptosis in myeloma cell lines resistant to steroids and 2-chlorodeoxyadenosine (2-CdA). AB - The paucity of effective cytotoxic agents for the treatment of steroid resistant multiple myeloma explains the ongoing search for alternative substances for chemotherapy of this disease. In the present study, the purine antagonist 2 chlorodeoxyadenosine (2-CdA, cladribine) and the pyrimidine antagonist 2',2' difluorodeoxycytidine (gemcitabine) were tested on four myeloma cell lines (i.e., U 266, OPM 2, RPMI 8226, IM 9), one plasma cell leukemia cell line (HS Sultan) and a myeloid control cell line (HL 60), all of which are resistant to 10-6 M dexamethasone. Gemcitabine has been found to be promising in the chemotherapy of other tumors with low proliferative activity, but its effectiveness against myeloma cells has not been analyzed so far. In our tests, gemcitabine induced a significant degree of apoptosis in all cell lines investigated. After incubation for 48 h with 10 microM gemcitabine, the median numbers of apoptotic cells were in the range of 45% in the OPM 2 and 79% in the U 266 cell line. All of the investigated cell lines were responsive to concentrations of 10 microM gemcitabine even after an exposure of only 30 min, three of them (U 266, HS Sultan, IM 9) also responded to a concentration of 10 nM. Higher concentrations and longer exposure times were necessary to suppress the growth of normal hematopoietic bone marrow progenitor cells. In contrast to gemcitabine, standard concentrations of 2-CdA (i.e., 30 and 300 nM) failed to induce a significant degree of apoptosis in the cell lines investigated but inhibited the growth of myeloid progenitor cells. The results suggest that gemcitabine induces apoptosis in myeloma and plasma cell leukemia lines resistant to steroids and 2-CdA. The fact that tumor cell apoptosis was achieved at concentrations clinically achievable and tolerable, which at the same time do not inhibit the growth of normal CFU-GM progenitor cells, favors the initiation of phase I trials with this drug for the treatment of multiple myeloma. PMID- 8724702 TI - Macrophage colony-stimulating factor levels in the plasma of bone marrow aspirate in several hematological malignancies. PMID- 8724703 TI - Pathogenesis of drug-induced gingival overgrowth. A review of studies in the rat model. AB - Drug-induced gingival overgrowth is a side effect associated principally with 3 types of drugs: anticonvulsant (phenytoin), immunosuppressant (cyclosporine A), and various calcium channel blockers (nifedipine, verapamil, diltiazem). In this review, we describe the features of phenytoin-, cyclosporine A- and nifedipine induced gingival overgrowth in rats and discuss factors influencing the onset and severity of these disorders. There are several features common to the gingival overgrowth induced by these drugs: 1) gingival overgrowth is more conspicuous in the buccal than in the lingual gingiva and less severe in the maxilla than in the mandible; 2) once the blood concentration of the drug reaches a certain level as a result of increasing the dose, the incidence of gingival overgrowth is 100% and its severity is dependent on the blood level, the most severe overgrowth being induced by cyclosporine A; 3) the duration of drug administration for maximal gingival overgrowth to develop is about 40 days; 4) the gingival overgrowth regresses spontaneously after discontinuing the drug; 5) accumulation of dental plaque is not essential for the onset of overgrowth, but plays a role in its severity; and 6) more severe overgrowth is induced in young than in old rats. Furthermore, male rats are more susceptible than females to nifedipine-induced gingival overgrowth. These results suggest that drug-induced gingival overgrowth in rats is dependent on the oral drug dose, blood drug level, age, and sex and that preexisting gingival inflammation is a factor relevant to its severity. Since these factors have also been suggested to be important determinants for human drug-induced gingival overgrowth, the rat model may prove valuable in the future for elucidating the molecular pathogenesis of the disorder. PMID- 8724704 TI - pH changes observed in the inflamed gingival crevice modulate human polymorphonuclear leukocyte activation in vitro. AB - Previous studies have noted a positive correlation between gingival inflammation and crevicular pH, which reportedly varies from 6.5 to 8.5. In the present study, we characterized the manner in which deviation from the "physiological" pH of blood (7.2) influences activation of chemotaxis, phagocytosis, superoxide generation, and degranulation by human polymorphonuclear leukocytes (PMNs). Purified PMNs were suspended in HEPES-buffered balanced salts solutions adjusted to pH 6.7, 7.2, 7.7, or 8.2. In a modified Boyden chamber, the chemotactic response to fMet-Leu-Phe was maximal at pH 7.2. In comparison, chemotaxis was significantly depressed at pH 7.7 and pH 8.2 (P < 0.05), but was not significantly different at pH 6.7. Activation of the respiratory burst by fMet Leu-Phe was optimal at pH 7.2, but was significantly depressed at pH 6.7 and 8.2 (P < 0.05). pH had little effect on N-acetyl-beta-glucosaminidase release from primary granules. However, lactoferrin release from the secondary granules of fMet-Leu-Phe-activated PMNs was significantly lower at pH 7.2 than at pH 6.7 or 8.2 (P < 0.05). Moreover, phagocytosis of opsonized bacteria was significantly lower at pH 7.2 than at pH 7.7. In addition to these effects on functional activation, extracellular pH influenced the magnitude of intracellular Ca2+ mobilization. Peak fMet-Leu-Phe-induced Ca2+ levels were significantly higher at pH 8.2 than at pH 7.2 (P < 0.01). These findings suggest that the pH of the periodontal environment can selectively influence PMN activation, thereby altering the balance between bacteria and the host response. PMID- 8724705 TI - The effect of periodontal treatment on periodontal bacteria on the oral mucous membranes. AB - The aim of this study was to investigate the effect of conventional periodontal treatment on the prevalence of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Prevotella intermedia on oral mucous membranes in patients with periodontitis. Fifteen patients were selected with moderate to severe periodontitis (mean age: 39.8 years, range: 26 to 58). From each patient the 4 deepest sites with bleeding on probing were selected for clinical and microbiological evaluation. At baseline, 6 weeks after oral hygiene instruction and extensive scaling and root planing, and 3 months after periodontal surgery, clinical parameters and microbiological samples including saliva, oral mucous membranes, and supra- and subgingival plaque were evaluated for the presence of the 3 test bacteria using indirect immunofluorescence. All clinical variables, except redness, showed a significant improvement after surgery. After treatment, the subgingival prevalence of the 3 putative periodontal pathogens had decreased significantly. However, almost no concomitant reduction in the prevalence of the bacteria was seen on the oral mucous membranes. This suggests that the oral mucous membranes may serve as a source for reinfection of the periodontium after treatment. PMID- 8724706 TI - A comparison of chlorhexidine, cetylpyridinium chloride, triclosan, and C31G mouthrinse products for plaque inhibition. AB - There are a large number of mouthrinse products available to the general public for use as adjuncts to oral hygiene. Many have not been evaluated and relatively few comparisons of products have been made. This study compared 4 mouthrinse products containing cetylpyridinium chloride (CPC), chlorhexidine, C31G, or triclosan with saline rinse included as a placebo control. Twenty dentate volunteers took part in this 4-day plaque regrowth study which had a single blind, randomized cross-over design balanced for residual effects. On day 1 of each study period, volunteers were rendered plaque free by a professional prophylaxis, suspended normal oral hygiene measures, and rinsed twice daily for 1 minute with 15 mL of the allocated rinse. On day 5, subjects were scored for disclosed plaque by plaque index and plaque area. By both measures the order of decreasing product efficacy was chlorhexidine, CPC and triclosan, C31G, and saline. All the differences in favor of the chlorhexidine product were highly significant as were those in favor of the other rinses compared to saline. It is concluded that the findings of this study reflect the actual chemical benefits of the products divorced from the indeterminate variable of toothbrushing. PMID- 8724707 TI - In vitro permeability evaluation and colonization of membranes for periodontal regeneration by Porphyromonas gingivalis. AB - The use of membranes for periodontal regeneration is well established. In clinical use, the exposure of membranes to the oral microflora may result in a pathway for periodontal infections. An important role in this process is played by Porphyromonas gingivalis. The purpose of the present study was to examine the colonization of 6 different bioresorbable and nonresorbable membranes for periodontal regeneration by the strain DSM 20709 of P. gingivalis and to determine the time needed by this microorganism to pass through the membranes. A device consisting of a tube sealed with the membranes and filled with a medium suitable for the growth of P. gingivalis was incubated in a bigger tube containing the same medium to study the process of colonization and the crossing of membranes. The outer tube was inoculated with 10(4) cells of P. gingivalis DSM 20709. The passage of bacteria through the membranes was monitored at 6, 24, and 48 hours by counting the number of cells in the inner tube. The colonized membranes were observed using a scanning electron microscope. Differences in the behavior of the 6 membranes analyzed were demonstrated. PMID- 8724708 TI - Analytical performance of an immunologic-based periodontal bacterial test for simultaneous detection and differentiation of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Prevotella intermedia. AB - The analytical performance of a membrane-based immunoassay for the simultaneous detection and differentiation of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Prevotella intermedia (including Prevotella nigrescens) was investigated. Positive reactions were observed for 71 of 71 reference strains and recent oral isolates of A. actinomycetemcomitans, P. gingivalis, and P. intermedia. No cross-reactivity was observed with 39 other common oral and environmental species. The specificity of the test was unaffected by the presence of potential oral interferents including whole blood, white blood cells, mucin, saliva, toothpastes, and oral rinses. A proficiency test by dental professionals using a standardized set of unknown simulated samples yielded a sensitivity of 97% (116/120) and a 100% specificity (240/ 240). An additional group including dental professionals and high school students was shown to be 99% proficient (1385/1397) in distinguishing proper from improper test function when processing control samples with normal test devices and devices with simulated error conditions. Comparisons to a culture standard for 104 subgingival plaque samples collected from 26 adult periodontitis patients yielded > 98% specificity for each of the test bacteria. In addition, the detection threshold for the test was determined to be equivalent to 10(4) cultivable test bacteria when compared to the culture standard. The data indicate that this membrane immunoassay is a valid and easy-to-use method for the detection of A. actinomycetemcomitans, P. gingivalis, and P. intermedia in subgingival plaque, at levels above the detection threshold of the test. PMID- 8724709 TI - The "cyclic" regimen of low-dose doxycycline for adult periodontitis: a preliminary study. AB - Specially-formulated low-dose doxycycline (LDD) regimens have been found to reduce collagenase activity in the gingival tissues and crevicular fluid (GCF) of adult periodontitis subjects in short-term studies. In the current, double-blind, placebo-controlled study, adult periodontitis patients were administered for 6 months a "cyclical" regimen of either LDD or placebo capsules; and various clinical parameters of periodontal disease severity, and both collagenase activity and degradation of the serum protein, alpha 1-PI, in the GCF were measured at different time periods. No significant differences between the LDD- and placebo-treated groups were observed for plaque index and gingival index. However, attachment levels, probing depth, and GCF collagenase activity and alpha 1-PI degradation were all beneficially and significantly (P < 0.05) affected by the drug regimen. We propose: 1) that LDD inhibits tissue destruction in the absence of either antimicrobial or significant anti-inflammatory efficacy; and 2) that long-term LDD could be a useful adjunct to instrumentation therapy in the management of the adult periodontitis patient. PMID- 8724710 TI - Detection of local and systemic cytokines in adult periodontitis. AB - Periodontitis is a chronic inflammatory disease of the soft and hard supporting tissues of the teeth and is a major cause of tooth loss in adults. The local host response to periodontopathic bacteria results in the release of inflammatory mediators and cytokines. Since cytokines are indicative of effector functions, we compared the pattern of cytokine production in periodontal patients and healthy controls. Specifically, we investigated the simultaneous presence of cytokines produced by T helper (Th)1, Th 2, and inflammatory cells which could be involved in periodontitis. We also compared the expression of these cytokine mRNAs in healthy and diseased tissues. No significant differences were detected at the protein or mRNA levels of the cytokines in the systemic circulation of patients and controls. The surface markers CD16 and CD56 were expressed on significantly fewer peripheral mononuclear cells of patients when compared to controls. gamma delta + T cells were found in half of the diseased tissues, but in none of the healthy tissues of either patients or controls. Finally, significant differences were observed between healthy and inflamed gingival tissues in the cytokine mRNA profile. Expression of IL-6 and IFN-alpha mRNA was significantly higher in diseased tissues compared to healthy tissues in patients. PMID- 8724711 TI - Long-term stability of Class II furcation defects treated with barrier membranes. AB - The present longitudinal study was designed to explore the long-term efficacy of guided tissue regeneration (GTR) in Class II furcation defects and establish the factors that might be responsible for modifying this response. Subjects with two or more mandibular molars, one of which had Class II furcation defects, received the hygienic phase of therapy followed by baseline clinical measurements and subgingival plaque sampling. GTR procedure was performed in furcation defect sites using expanded polytetrafluoroethylene (ePTFE) membranes, while the other non-furcated molars received only scaling and root planning. Twenty-eight subjects (13 females, 15 males) aged 27 to 66 were included in this longitudinal analysis. Post-surgical treatment included routine home care supplemented with daily chlohexidine rinse and systemic tetracycline. Membranes were retrieved 4 to 6 weeks after surgery. During the first year, patients were initially seen bi weekly and subsequently monthly for professional prophylaxis. At the end of this year, clinical measurements and samples were obtained. For the next 2 years, patients were seen bi-annually for maintenance visits. Clinical measurements and microbiological samples were then repeated. Next, a tighter maintenance protocol was established and patients were seen quarterly for scaling and oral hygiene reinforcement. Final measurements and samples were taken again 1 year later (4 years postoperative). Significant probing reduction (3.00 mm) and gain in horizontal attachment (2.59 mm) were obtained 1 year postsurgery for the GTR sites. These changes were maintained over 4 years with a slight decline at the end of year 3. Changes in probing depth (PD) from year 1 to 4 served to dichotomize the sites into stable (delta PD < or = 0.9 mm), and unstable (PD increase > or = 1 mm). Of the 54 sites available for this analysis only 5 (9.3%) were unstable while 49 (90.7%) were stable or even further improved. Sites which exhibited minimal or no plaque (plaque index [PI] < or = 1) over the tight maintenance period had a further decrease in mean probing depth (0.43 mm) compared with a slight increase (-0.06 mm) in mean probing depth in sites with PI > or = 2 mm (P = 0.0235). The same phenomenon was observed for changes in relative attachment level (RAL): mean gain in RAL was 0.61 mm compared to 0.25 mm for the 2 groups, respectively (P = 0.07). Actinobacillus actinomycetemcomitans was only isolated from 2 sites at year 3, and none at year 4, compared to 21.45% of the sites at baseline. Porphyromonas gingivalis positive sites showed a continual decline over the years: 14.28% at baseline, 10.71% at year 1, and 5.1% at year 4. On the contrary, Prevotella intermedia (Pi) and Bacteroides forsythus (Bf) infected sites remained at approximately the same rate throughout the 4 years of the study (40% to 50% and 30% to 40% for Pi and Bf, respectively). Of these, Pi-infected sites exhibited less favorable clinical results compared to sites which were not infected with this microorganism. In summary, furcation defects treated with membrane barriers can be maintained in health for at least 4 years; however, good oral hygiene and frequent recall visits as part of a complete anti-infective therapy are essential. Finally, once treated, these teeth are comparable to similar molar teeth with no previous history of furcation pathosis. PMID- 8724712 TI - Human papillomavirus in a patient with severe gingival overgrowth associated with cyclosporine therapy. A case report. AB - Cyclosporin a is an endecapeptide that has been used clinically since 1978 as an immunosuppressant agent. Although cyclosporine appears to be uniformly beneficial in the treatment of a wide variety of disorders, its use may result in a number of side effects. One of the most important adverse effects is gingival overgrowth. This report relates a case of severe gingival overgrowth in a 31-year old female who had received cyclosporine therapy in conjunction with a kidney transplant. Because of the severe gingival overgrowth, we analyzed a biopsy, which was positive for human papillomavirus. The case was treated and followed for 18 months. PMID- 8724713 TI - Free gingival graft following biopsy: a case report of tissue management. AB - Excisional biopsies require wide margins to facilitate removal of diseased tissue. Such margins often result in soft-tissue defects. These defects can cause reduced esthetics, ineffective oral hygiene performance, and postsurgical root sensitivity. This case documents the combination of two dental procedures: biopsy and the free gingival graft. A pyogenic granuloma, as diagnosed by histologic report, was removed by excisional biopsy and the resultant defect repaired by using a free gingival graft. Palatal tissue was grafted immediately to the site of the biopsy and sutured in place to cover the loss of attached tissue and papilla. At 6 months, the graft appeared to restore gingival health and maintain both esthetics and function in the surgical biopsy site. The patient was well served by correcting the resultant biopsy defect in a one-step procedure which encouraged healing and an excellent esthetic result. PMID- 8724714 TI - Dental considerations for a Glanzmann's thrombasthenia patient: case report. AB - Glanzmann's thrombasthenia is a qualitative platelet disorder characterized by a deficiency in the platelet membrane glycoproteins (GP) IIb-IIIa. It belongs to a group of hereditary platelet disorders typified by normal platelet numbers and a prolonged bleeding time. The bleeding seen in Glanzmann's thrombasthenia usually includes bruising, epistaxis, gingival hemorrhage, and menorrhagia. Spontaneous, unprovoked bleeding is unusual. The severity of bleeding is unpredictable in thrombasthenia and does not correlate with the severity of the platelet GP IIb IIIa abnormality. The present case report describes the dental treatment of a patient with Glanzmann's thrombasthenia. A 39-year-old female with a history of Glanzmann's thrombasthenia presented for periodontal therapy for spontaneous gingival hemorrhage. The patient had been sporadically seen in the past and had a record of only returning for appointments on an "emergency" basis. The periodontal findings revealed a diagnosis of moderate to advanced adult periodontitis in all quadrants. After all dental options had been discussed, the treatment of choice was determined to be extraction of the remaining dentition and fabrication of immediate dentures. The patient received a loading dose of 5 grams of aminocaproic acid (EACA) intravenously 3 hours prior to the surgery. At the beginning of the extractions 1 gram of EACA per hour continuous infusion and a 6 pack of platelets was administered. The patient tolerated the extractions well. All sites healed normally. The patient has had no difficulty in adjusting to the dentures. The case report discusses a possible treatment option in a noncompliant patient having Glanzmann's thrombasthenia and briefly discusses other hereditary bleeding disorders with similar presentations. PMID- 8724715 TI - Report of a lateral periodontal cyst and gingival cyst occurring in the same patient. AB - The lateral periodontal cyst and gingival cyst of the adult are both developmental odontogenic cysts located predominantly in the mandibular premolar canine region. In this paper, a case report is described of a lateral periodontal cyst and gingival cyst occurring in the same patient along with a discussion of clinical, radiographic, and histological features of these lesions. PMID- 8724716 TI - The potential role of growth and differentiation factors in periodontal regeneration. AB - Polypeptide growth and differentiation factors (GDFs) are a class of biological mediators which have been shown to play a critical role in the stimulation and regulation of the wound healing process. Growth factors involved in repair and regeneration regulate several key cellular processes such as mitogenesis, chemotaxis, differentiation, and metabolism. The sequence of events necessary for periodontal regeneration relies on the above processes for osteogenesis, cementogenesis, and connective tissue formation. Numerous preclinical in vitro and in vivo studies have demonstrated that certain growth factors modulate putative components of periodontal wound healing resulting in substantial regeneration of the periodontium in animals. However, these therapies are currently considered to be in the experimental stage of development and no growth factor therapy to treat periodontitis in humans has received approval by the Food and Drug Administration (FDA). This informational paper reviews the various aspects of GDFs on cells and tissues involved in periodontal wound healing. This paper was prepared by the Committee on Research, Science and Therapy of The American Academy of Periodontology and is intended for members of the dental profession. PMID- 8724717 TI - Effects of maternal smoking on amniotic fluid volume and fetal urine output. AB - The purpose of this study was to assess the effects of maternal smoking on amniotic fluid volume and fetal urine output. Seventeen women (10 smokers and 7 non-smokers) were serially assessed through pregnancy. Smoking status was determined by self-report and carbon monoxide manometry. Amniotic fluid volume was assessed by sonographically measuring the maximum vertical pocket, the amniotic fluid index, and the two dimensions of the largest pockets. The coronal area formula was used to calculate bladder volume at 5-minute intervals for 20 minutes. Urine output was calculated from serial measurements of bladder volume. The mean gestational age of the first session was 26.8 (2.7) weeks, of the second session 32.4 (1.6) weeks, and of the last session 36.9 (2.1) weeks. In the univariate analysis there were no differences between smokers and non-smokers at any of the three visits for gestational age, fetal weight, maximum vertical pocket, amniotic fluid index, two-dimensional maximum pocket, and urine output. In the regression model, while controlling for gestational age, smoking status could not predict significant differences in any parameter. Maternal smoking does not appear to affect either amniotic fluid volume or fetal urine output. PMID- 8724718 TI - Idiopathic rapidly progressive (crescentic) glomerulonephritis in pregnancy. AB - Idiopathic rapidly progressive glomerulonephritis is an uncommon kidney disorder characterized by the rapid deterioration of renal function in association with glomerular crescent formation on renal biopsy. End-stage kidney failure is often the final outcome. There is only one complete case report of this condition during pregnancy. We describe a patient who initially presented with classic signs and symptoms of severe preeclampsia. She delivered a premature neonate that ultimately did well, but the mother subsequently demonstrated incremental deterioration in renal function. A kidney biopsy documented crescentic glomerulonephritis, and her clinical course was consistent with rapidly progressive glomerulonephritis. PMID- 8724720 TI - Prenatal diagnosis of a choledochal cyst: a case report and review of the literature. AB - There are 16 reported cases of prenatally diagnosed choledochal cyst in the literature. We present a new case diagnosed at 29 weeks' gestation by routine ultrasound scanning. At 40 weeks' gestation, a male infant was born by spontaneous delivery. At 16 weeks of age, the patient underwent a laparotomy, which confirmed the diagnosis of a choledochal cyst. The cyst and the gallbladder were removed en bloc and reconstruction of the biliary tree was performed by formation of a retrocolic Roux-en-Y-hapatojejunostomy with an antireflux valve. Histological examination of the cyst showed a thickened fibrous tissue wall with necrotising areas and without epithelial lining. The patient's postoperative course was uneventful. Review of the literature shows that it is possible to make a presumptive prenatal diagnosis of this anomaly as early as 15 weeks' gestation. The differential diagnosis of a sonolucent lesion in the fetus should also include the rare choledochal cyst. Treatment of choice is the radical excision of the cyst with construction of a Roux-en-Y hepaticoenterostomy. Long-term results with this method are very good. Surgery may be instituted at the earliest possible opportunity before severe complications such as cholangitis, liver abscesses, cirrhosis, and malignant degeneration can occur. PMID- 8724719 TI - Prenatal detection of the separation of the great toe, toe syndactyly, and large bilateral choroid plexus cysts in a fetus with trisomy 18. AB - Prenatal sonographic presentation of toe deformities is associated with fetal aneuploidies. This report presents a second-trimester fetus with large bilateral choroid plexus cysts, clenched hands, separation of the great toe, toe syndactyly, abnormal double maternal serum biochemical screening results, and trisomy 18. We suggest a careful ultrasound screening of the fetal limbs and other organs once a choroid plexus cyst has been identified. If abnormal sonographic findings are present, or if the results of the maternal serum biochemical screening are abnormal, karyotyping should be recommended. PMID- 8724721 TI - Alloimmune thrombocytopenia of the newborn: neurodevelopmental sequelae. AB - Alloimmune thrombocytopenia of the newborn (AITN) is due to the transplacental passage of maternal antibodies directed against fetal platelet antigens, most commonly PLA, (HPA-1a). Sensitization and clinical manifestations of disease can occur in the first pregnancy. It carries a mortality rate of 15%, usually due to intracranial hemorrhage (ICH). Twenty-four cases of AITN were reviewed, five of whom had radiological evidence of ICH in utero. Petechiae and/or bruising were present in 14 babies. Platelet counts at birth ranged from 5 to 206 x 10(9)/L. One infant died. The duration of follow-up for the four survivors of ICH ranged from 15 to 71 months. All had serious neurodevelopmental sequelae, including severe mental retardation, cortical blindness, seizures, and cerebral palsy. The emotional and financial cost created by the care of these children is immeasurable. Steps to identify and prevent AITN should be part of routine prenatal care if we are to reduce the morbidity and mortality caused by this disorder. PMID- 8724722 TI - Use of tissue type plasminogen activator in neonates: case reports and review of the literature. AB - Recombinant tissue type plasminogen activator (rt-PA) has been used successfully in neonates for resolution of thrombotic processes, both arterial and venous. We report on 2 neonates with vena cava inferior thrombosis and with biochemical evidence of liver dysfunction in whom rt-PA treatment had to be interrupted because of significant bleeding tendency, without complete resolution of the thrombus. Therapy failure may be due to the age of the thrombus and the systemic way of administration, bypassing the site of the thrombus by collateral circulation. Impaired liver function may result in decreased degradation of rt-PA with prolonged effect and higher risk for bleeding complications. We suggest that lower doses should be administered in these patients. PMID- 8724723 TI - Congenital hernia of the cord and associated patent omphalomesenteric duct: a frequent neonatal problem? AB - Six neonates with hernia of the umbilical cord (HUC) and associated patent omphalomesenteric duct (POMD) were treated in our hospital in a 10-year period and are reported in order to emphasize the potential hazard of clamping the bowel at the time of the handling of the umbilicus. Any unusual thickening of the base of the cord along with even the most minute fistula opening to its side should alert the physician to the existence of these combined anomalies. The cord in such patients must be clamped a safe distance away and early pediatric surgical consultation must be contemplated. Obstetricians, pediatricians, and nurses who customarily clamp, manipulate, or shorten the cord in the delivery room or upon arrival in the nursery should be aware of this not-so-rare combination of anomalies of the umbilical cord and the intestines. PMID- 8724724 TI - Umbilical plasma erythropoietin correlations with blood gases and gestational age in appropriately grown infants. AB - Erythropoietin (EPO) levels were measured using an ELISA method in umbilical cord plasma from 68 appropriately grown neonates at 27 to 43 weeks of gestation (EGA). A consistent but small (5%) difference was found between the EPO levels in the umbilical artery (Ua), 21.0 +/- 2.5 mlU/mL (mean +/- SEM), and umbilical vein (Uv), 22.0 +/- 2.7 mlU/mL (p < 0.02, n = 30). Significant inverse correlations were found between Ua EPO levels and pO2 (r = -0.44, p < 0.002), pH (r = -0.68, p < 0.0001), as well as base deficit (r = -0.56, p < 0.0001). A significant correlation was found between EGA and Ua EPO from 27 to 43 weeks of gestation (r = 0.45, p < 0.001, n = 68). One and 5 minute Apgar scores did not correlate with EPO. These findings indicate that EPO correlates strongly with cord gas parameters and thus may serve as a clinically useful marker for both subacute fetal distress and chronic uteroplacental insufficiency. PMID- 8724725 TI - Amniotic fluid index and perinatal morbidity. AB - Our purpose was: (1) to determine whether assessment of amniotic fluid index in high risk patients with a reactive nonstress test (NST) allowed improved recognition of the fetus at risk for perinatal morbidity than a reactive NST alone; and (2) to determine the optimal low amniotic fluid index (AFI) which should prompt clinical concern. The last NST performed within a week of delivery and amniotic fluid index were retro-spectively compared with various indices of perinatal morbidity. In fetuses with a reactive NST, decreasing AFI was directly correlated with a risk of 5-minute Apgar score of less than 7 and delivery for fetal distress. When various subgroupings of AFI were compared, 7 cm or greater appeared to have a better inverse correlation with the indices of morbidity than lower cut-off values. The addition of AFI assessment to the standard NST allows better prediction of perinatal morbidity than the NSTs alone. Seven centimeters appears to be a reasonable cut-off for clinical concern. PMID- 8724726 TI - Evaluation of suprapubic bladder aspiration for detection of group B streptococcal antigen by latex agglutination in neonatal urine. AB - Our objective was to determine whether urine collection by suprapubic bladder aspiration (SBA) improves the specificity of the group B streptococcal (GBS) latex agglutination (LA) test by avoiding contamination of urine with GBS from perineal and rectal colonization that can result in a positive LA test in an uninfected infant when the urine is collected by bag. Part 1 consists of a retrospective review of the medical records of 113 infants who had urine collected by SBA for GBS LA testing as part of evaluation for possible sepsis. The sensitivity and specificity of the urine LA test was assessed by comparing it with blood culture results. In part 2, a prospective analysis was performed of 19 newborns who had rectal and vaginal/penile cultures as well as urine by SBA and bag for GBS cultures and LA. Results of LA testing on urine collected by both of these methods were compared with results of urine, perineal, and rectal cultures. In the retrospective review of GBS LA testing performed on 113 consecutive urine specimens collected by SBA from neonates being evaluated for suspected sepsis, the sensitivity and specificity were 67% and 89%, respectively, when compared with blood culture results. Twelve infants who had a positive LA test result but a sterile blood culture (BC-,LA+) were compared with 95 infants with both blood cultures and urine LA tests negative for GBS (BC-, LA-). BC-, LA+ infants were more likely than those with BC-, LA- to have an immature to total neutrophil (I/T) ratio > or = 0.16 at 12 and 24 hours (p = 0.04 and 0.02, respectively). In the prospective study, we found that a positive GBS LA test can be due to perineal contamination and possibly to gastrointestinal absorption of GBS antigen. No false positive LA test results occurred on urine obtained by SBA; however, use of this method failed to detect the one infant with GBS bacteremia. Because of suboptimal sensitivity and specificity, use of the GBS LA test on urine obtained either by SBA or bag cannot be recommended for diagnosis of early onset GBS disease. PMID- 8724727 TI - Right diaphragmatic eventration simulating a congenital diaphragmatic hernia. AB - We describe an infant with severe eventration of the right diaphragm and pulmonary hypoplasia who presented like a newborn with congenital diaphragmatic hernia complicated by persistent pulmonary hypertension. Surgical correction while on extracorporeal life support was unsuccessful due to attachments of the liver which prevented reduction into the abdominal cavity and our inability to distinguish the true defect from complete agencies of the right hemidiaphragm. At autopsy the pulmonary remnant and the fibrous membrane separating it from the liver were identified. PMID- 8724728 TI - Fibrin glue treatment for recurrent pneumothorax in a premature infant. AB - A case of a premature infant in whom recurrent pneumothorax was successfully treated by injection of fibrin glue into the pleural cavity through the chest tube is reported. Pleurodesis with fibrin glue was found to be both safe and effective. PMID- 8724729 TI - Lessons learned from the non-obese diabetic mouse II: Amelioration of pancreatic autoimmune isograft rejection during pregnancy. AB - To determine whether pregnancy provides an improved milieu for fetal/neonatal pancreas/islet transplantation, we studied neonatal pancreatic implants into non obese diabetic (NOD) female mice during early gestation. We monitored maternal glycemic status, birthweight of the offspring, and graft histology to assess the efficacy of transplantation. One hundred and thirteen twelve-week-old NOD female mice were randomized into four groups as follows: (1) non-pregnant NOD mice received a sham operation; (2) non-pregnant NOD mice received neonatal pancreatic transplants; (3) pregnant NOD mice received a sham operation; and (4) pregnant NOD mice received neonatal pancreatic transplants. Pancreas segments from 3 neonatal NOD mice were placed via an incision 1 to 2 mm distal to the ear-skull junction of each of the recipients. Maternal blood glucose and glycated hemoglobin were determined between days 18 and 20 after the surgery. Pups were weighed within 5 to 6 hours after delivery. Pregnant NOD that received transplants (n = 29) had lower glucose and glycated hemoglobin (GHb) than sham operated pregnant controls (n = 26) (4.9 +/- 0.05 versus 9.0 +/- 5.0 mmol/L, p < 0.001 for glucose and 2.0 > or = 0.2 versus 3.0 > or = 1.2%, p < 0.008 for GHb) at 18 to 20 days of gestation. Controlling for litter size showed a decrease in birthweight for offspring of transplant recipients versus offspring of pregnant controls (1.59 +/- 0.08 versus 1.65 +/- 0.08 g, p < 0.002). Histological scoring of transplanted tissue at day 21 indicated that the lymphocytic infiltration in the pregnant group was significantly less than the control group (2.9 +/- 1.2 versus 4.9 +/- 0.2, p < 0.0001). We conclude that the pregnant NOD mouse provides a useful transplant model, that pregnancy provides an opportunity to increase beta-cell mass with transplanted tissue, and that pancreatic transplantation decrease birthweight and macrosomia in the offspring of NOD mice. PMID- 8724730 TI - Relationship between erythrocyte deformability and hematocrit in maternal and cord blood. PMID- 8724731 TI - Professor Robley D. Evans 1907-1995. PMID- 8724732 TI - Three-dimensional reconstruction of vascular trees: experimental evaluation. AB - This paper is the second of two that together present a novel approach to the problem of reconstructing vascular trees from a small number of projections. Previously, we described the reconstruction algorithm and how it effectively circumvents the matching or "correspondence problem" found in most photogrammetric or computer-vision-based approaches. The algorithm is fully automatic and assumes that the imaging geometry is known, the vascular tree is a connected structure, and that its center-lines have been identified in three or more images. It employs consistency and connectivity constraints and comprises three steps: The first generates a connected structure representing the multiplicity of solutions that are consistent with the first two views; the second assigns a measure of agreement to each branch in this structure based on one or more additional projections; and the third step employs this measure to distinguish between those branches comprising the vasculature and the accompanying artifacts. This paper addresses the issue of validation via simulations and experiments. In addition to a clinical case, we examine the performance of the algorithm when applied to simulated projections of two 3-D vascular models, both representative of the complexity faced in coronary and cerebral angiography. The results in each instance are impressive and demonstrate that adequate reconstructions may be obtained with as few as three distinct views. PMID- 8724733 TI - Tissue-phantom ratios from percentage depth doses. AB - When converting fractional (percentage) depth doses to tissue-phantom ratios, one must use a factor that accounts for the different source-to-point distances. Two minor correction factors are also involved. One is the ratio of total to primary dose at the two different distances from the source, for the same depth and field size. This factor is usually ignored. It was determined experimentally that this can introduce up to 1.5% error at 6 MV. The second correction factor reflects differences related to scattered photons and electrons at the depth of normalization in the two geometries. This correction is accounted for in published conversion procedures. It was found to be less than 1% provided the normalization depth is sufficient for electron equilibrium, which occurs first well beyond the depth of maximum dose. One may avoid electron-equilibrium problems by using an interim normalization depth that provides electron equilibrium with some margin, renormalizing to a shallower depth if desired. With this precaution, the accuracy when measured fractional depth doses were converted to tissue-phantom ratios was comparable to that of directly measured tissue phantom ratios even when the correction factors were ignored. PMID- 8724734 TI - Testing of dynamic multileaf collimation. AB - It has been shown that intensity-modulated fields have the potential to deliver optimum dose distributions, i.e., high dose uniformity in the target and lower doses in the surrounding critical organs. One way to deliver such fields is by using dynamic multileaf collimation (DMLC). This capability is already available in research mode on some treatment machines. While much effort has been devoted to developing algorithms for DMLC, the mechanical reliability of this new treatment delivery mode has not been fully studied. In this work, we report a series of tests designed to investigate the mechanical aspects of DMLC and their implications on dosimetry. Specifically, these tests were designed to examine (1) the stability of leaf speed, (2) the effect of lateral disequilibrium on dose profiles between adjacent leaves, (3) the significance of acceleration and deceleration of leaf motion, (4) the effect of positional accuracy and rounded end of the leaves, and (5) create a simple test pattern that may serve as a basis for routine quality assurance checks. Results of these tests are presented. The implications on dosimetry and consideration for the design of leaf motion are discussed. PMID- 8724735 TI - Water calorimetry and ionization chamber dosimetry in an 85-MeV clinical proton beam. AB - In recent years, the increased use of proton beams for clinical purposes has enhanced the demand for accurate absolute dosimetry for protons. As calorimetry is the most direct way to establish the absorbed dose and because water has recently been accepted as standard material for this type of beam, the importance of water calorimetry is obvious. In this work we report water calorimeter operation in an 85-MeV proton beam and a comparison of the absorbed dose to water measured by ionometry with the dose resulting from water calorimetric measurements. To ensure a proper understanding of the heat defect for defined impurities in water for this type of radiation, a relative response study was first done in comparison with theoretical calculations of the heat defect. The results showed that pure hypoxic water and hydrogen-saturated water yielded the same response with practically zero heat defect, in agreement with the model calculations. The absorbed dose inferred from these measurements was then compared with the dose derived from ionometry by applying the European Charged Heavy Particle Dosimetry (ECHED) protocol. Restricting the comparison to chambers recommended in the protocol, the calorimeter dose was found to be 2.6% +/- 0.9% lower than the average ionometry dose. In order to estimate the significance of chamber-dependent effects in this deviation, measurements were performed using a set of ten ionization chambers of five different types. The maximum internal deviation in the ionometry results amounted to 1.1%. We detected no systematic chamber volume dependence, but observed a small but systematic effect of the chamber wall thickness. The observed deviation between calorimetry and ionometry can be attributed to a combination of the value of (Wair/e)p for protons, adopted in the ECHED protocol, the mass stopping power ratios of water to air for protons, and possibly small ionization chamber wall effects. PMID- 8724736 TI - Evaluation of bremsstrahlung spectra generated by a 4-MeV linear accelerator. AB - A modified method for the evaluation of bremsstrahlung spectra on the basis of transmission measurements is presented. The method is tested for two 4 MeV therapy accelerators. The calculated spectra are compared with other experimental results and Monte Carlo simulation. PMID- 8724737 TI - A new radiotherapy surface dose detector:the MOSFET. AB - Radiotherapy x-ray and electron beam surface doses are accurately measurable by use of a MOS-FET detector system. The MOSFET (Metal Oxide Semiconductor Field Effect Transistor) is approximately 200-microns in diameter and consists of a 0.5 microns Al electrode on top of a 1-microns SiO2 and 300-microns Si substrate. Results for % surface dose were within +/- 2% compared to the Attix chamber and within +/- 3% of TLD extrapolation results for normally incident beams. Detectors were compared using different energies, field size, and beam modifying devices such as block trays and wedges. Percentage surface dose for 10 x 10-cm and 40 x 40-cm field size for 6-MV x rays at 100-cm SSD using the MOSFET were 16% and 42% of maximum, respectively. Factors such as its small size, immediate retrieval of results, high accuracy attainable from low applied doses, and as the MOSFET records its dose history make it a suitable in vivo dosimeter where surface and skin doses need to be determined. This can be achieved within part of the first fraction of dose (i.e., only 10 cGy is required.) PMID- 8724738 TI - Optimization of conformal electron beam therapy using energy- and fluence modulated beams. AB - Fluence modulation of multiple electron beams of various energies has been used to optimize the delivered dose distribution during electron beam radiation therapy. By maximizing the probability of achieving tumor control without causing severe complications electron beam fluence profiles have been optimized for superficial target volumes. It is possible to use several equiportal fluence modulated electron beams to modify the energy deposition with depth in a controlled manner making it possible to use the technique as an alternative to bolus. The technique was tested in two representative phantom geometries and in three clinical patient geometries using a set of five and two different energies. The local maxima in dose for the plans with five energies were typically lower than with the conventional or advanced bolus techniques. The principles for how the technique could be carried out in the future with a fourth generation radiotherapy accelerator are also indicated. PMID- 8724739 TI - Thermoluminescent dosimetry in electron beams: energy dependence. AB - The response of thermoluminescent dosimeters to electron irradiations depends on the radiation dose, mean electron energy at the position of the dosimeter in phantom, and the size of the dosimeter. In this paper the semi-empirical expression proposed by Holt et al. [Phys. Med. Biol. 20, 559-570 (1975)] is combined with the calculated electron dose fraction to determine the thermoluminescent dosimetry (TLD) response as a function of the mean electron energy and the dosimeter size. The electron and photon dose fractions, defined as the relative contributions of electrons and bremsstrahlung photons to the total dose for a clinical electron beam, are calculated with Monte Carlo techniques using EGS4. Agreement between the calculated and measured TLD response is very good. We show that the considerable reduction in TLD response per unit dose at low electron energies, i.e., at large depths in phantom, is offset by an ever increasing relative contribution of bremsstrahlung photons to the total dose of clinical electron beams. This renders the TLD sufficiently reliable for dose measurements over the entire electron depth dose distribution despite the dependence of the TLD response on electron beam energy. PMID- 8724740 TI - Measured electron energy and angular distributions from clinical accelerators. AB - Electron energy spectra and angular distributions, including angular spreads, were measured using magnetic spectrometer techniques, at isocenter, for two clinical linear accelerators: one scanning beam machine, which achieves field flatness by scanning a pencil beam over the desired field at the patient plane, and one scattering foil machine, which disperses the electrons through a graded thickness scattering foil. All measurements were made at isocenter (in the patient plane), in air, 1 m from the nominal accelerator source. The energy measurements were confined to electrons traveling along the central axis; any widely scattered electrons were effectively neglected. The energy spectra of the scanning beam machine are all of nearly Gaussian shape and energy full-width-at half-maximum intensity (FWHM) of about 5% of the peak mean energy (denoted (E0)*). The energy spectra of the scattering foil machine have a variety of forms as a function of energy, including even spectra with double peaks, and spectra which changed with time. The FWHM values ranged from 9%-22% of (E0)*. The angular spread measurements, at isocenter, yielded sigma theta (x) x (E0)* approximately 295 mrad-MeV for the scanning beam machine, and 346 mrad-MeV for the scattering foil machine, where sigma theta x denotes the standard deviation of the plane projected angular distribution. These angular spreads are 30%-40% smaller than angular spreads reported by others on a very similar machine using the penumbra method. Possible causes of this discrepancy are discussed. PMID- 8724741 TI - Systematic verification of a three-dimensional electron beam dose calculation algorithm. AB - A three-dimensional electron beam dose calculation algorithm implemented on a commercial radiotherapy treatment planning system is described. The calculation is based on the M. D. Anderson Hospital (M.D.A.H.) pencil beam model, which uses the Fermi-Eyges theory of thick-target multiple Coulomb scattering. To establish the calculation algorithm's accuracy as well as its limitations, it was systematically and extensively tested and evaluated against a set of benchmark measurements. Various levels of dose and spatial tolerances were used to validate the calculation quantitatively. Results are presented in terms of the percentage of data points meeting a specific tolerance level. The algorithm's ability to accurately simulate commonly used clinical setup geometries, including standard or extended SSDs, blocked fields, irregular surfaces, and heterogeneities, is demonstrated. Regions of disagreement between calculations and measurements are also shown. The clinical implication of such disagreements is addressed, and the algorithmic assumptions involved are discussed. PMID- 8724742 TI - A method for predicting the variation of the depth of maximum dose in shaped electron fields. PMID- 8724743 TI - Radiation therapy dosimetry using magnetic resonance imaging of polymer gels. AB - Further progress in the development of polymer gel dosimetry using MRI is reported, together with examples of its application to verify treatment plans for stereotactic radiosurgery and high dose rate brachytherapy. The dose distribution image produced in the tissue-equivalent gel by radiation-induced polymerization, and encoded in the spatial distribution of the NMR transverse relaxation rates (R2) of the water protons in the gel, is permanent. Maps of R2 are constructed from magnetic resonance imaging data and serve as a template for dose maps, which can be used to verify complex dose distributions from external sources or brachytherapy applicators. The integrating, three-dimensional, tissue-equivalent characteristics of polymer gels make it possible to obtain dose distributions not readily measured by conventional methods. An improved gel formulation (BANG-2) has a linear dose response that is independent of energy and dose rate for the situations studied to date. There is excellent agreement between the dose distributions predicted using treatment planning calculations and those measured using the gel method, and the clinical practical utility of MRI-based polymer gel dosimetry is thereby demonstrated. PMID- 8724744 TI - Monte Carlo simulation of 252CF dose distribution brachytherapy. AB - The calculation of absorbed dose rate of photons and neutrons for the linear source of 252Cf with the active length of 2 cm is presented. Monte Carlo code and neutron cross-section data processed from ENDF/B-6 library were used. The achieved values of the absorbed dose rates are in good agreement with those published by other authors. The results of the calculation are applied to the gynecological configuration used in brachytherapy. PMID- 8724745 TI - The application of transit dosimetry to precision radiotherapy. AB - A method of using electronic portal imaging (EPI) for transit dosimetry is described. In this method, a portal image of the treatment field is first aligned with a digitally reconstructed radiograph (DRR) to geometrically relate the computed tomography (CT) scan, used to generate the DRR, with the EPI. Then the EPI is corrected for scatter within the patient to yield a map of primary fluence striking the detector. This is backprojected through the planning CT data set to yield a distribution of primary fluence within the patient. This distribution is then convolved with dose deposition kemels to yield a map of dose delivery within the patient. Such a distribution may be compared with the dose distribution resulting from the original treatment plan in order to evaluate the adequacy of the treatment. This method has been evaluated using a humanoid phantom. We find the transit dosimetry relative dose distribution when compared with film and thermoluminescent dosimeter (TLD) measurements and compared with our planning system to agree within 2% in the pelvic region of a humanoid phantom. PMID- 8724746 TI - Angular-dependent coherent scatter measured with a diagnostic x-ray image intensifier-based imaging system. AB - Low-angle scatter of x rays at diagnostic energies is primarily coherent. This coherence gives rise to interference effects resulting in x-ray diffraction patterns that are characteristic of the scattering material. A method is described of imaging these low-angle (0 degree-10 degrees) x-ray diffraction properties of tissue specimens using a diagnostic x-ray beam and image intensifier-based system. The coherent-scatter cross sections of several materials measured this way are presented. It is shown theoretically that the measurements made with this system can be expressed as the mono-energetic cross section "blurred" by the x-ray spectrum using a linear superposition integral. Experimental results using aluminum powder confirm this. Using a 70 kVp x-ray beam filtered with gadolinium to reduce the spectral width, materials such as water, Lucite, and hydroxyapatite all have significantly different diffraction patterns. The cross sections determined from this analysis from the basis of a unique method of characterizing and identifying tissue samples according to their atomic structure rather than x-ray attenuation properties. PMID- 8724747 TI - Energy imparted and effective doses in computed tomography. AB - The radiation risk to patients undergoing computed tomography (CT) examinations may be characterized by using the dose descriptors of effective dose equivalent (HE) and effective dose (E). Values of HE and E, however, are much more difficult to obtain than the total energy imparted (epsilon) to the patient. In this study, representative values of the ratios HE/epsilon and E/epsilon were obtained using published Monte Carlo organ dose computations for typical CT x-ray spectra. Values of patient dose per unit energy imparted can be combined with independent estimates of energy imparted to quantify the dose to a patient undergoing any type of CT examination. PMID- 8724748 TI - Radiation damage of amorphous silicon, thin-film, field-effect transistors. AB - The effect of 60Co radiation on the noise and drain-source current characteristics of hydrogenated amorphous silicon (alpha-Si:H) field-effect transistors (FETs) was examined as a function of dose to cumulative doses as high as approximately 2 x 10(4) Gy. Following these measurements, room-temperature and elevated-temperature annealing of induced radiation damage was examined. The FETs examined are representative of those incorporated in alpha-Si:H arrays under development for various x-ray medical imaging applications. No significant effect upon the noise characteristics of the FETs was observed as a result of the radiation. The predominant drain-source current effect with increasing dose was a shift of the transfer characteristic toward negative gate voltage and/or a decrease of the transfer characteristic subthreshold slope. This resulted in large increases in leakage current for gate voltages where the FETs were initially highly nonconducting. This leakage current increase was less pronounced for more negative gate voltages and was further diminished by maintaining the FETs at a more negative gate voltage during the irradiation. Following the radiation measurements, room-temperature annealing resulted in a 10% to 50% reduction in the leakage current in the first day followed by a logarithmic decrease thereafter. Elevated-temperature annealing for 2 h at 200 degrees C restored FET leakage current and threshold voltage properties to their preirradiation values. The irradiation effects are small for cumulative doses less than approximately 100 Gy, which is larger than the clinical lifetime dose for an imaging detector for chest radiography or for fluoroscopy (with infrequent exposure to the direct beam). For significantly higher dose applications such as mammography, fluoroscopy (with frequent direct beam exposure), and radiotherapy imaging, the results suggest that periodic elevated-temperature annealing or operation of the arrays at more negative gate voltages may be necessary. PMID- 8724749 TI - Design of a laser scanner for a digital mammography system. AB - We have developed a digital readout system for radiographic images using a scanning laser beam. In this system, electrostatic charge images on amorphous selenium (alpha-Se) plates are read out using photo-induced discharge (PID). We discuss the design requirements of a laser scanner for the PID system and describe its construction from commercially available components. The principles demonstrated can be adapted to a variety of digital imaging systems. PMID- 8724750 TI - Radionuclide photon dose kernels for internal emitter dosimetry. AB - Photon point dose kernels and absorbed fractions were generated in water for the full photon emission spectrum of each radionuclide of interest in nuclear medicine, by simulating the transport of particles using Monte Carlo. The kernels were then fitted to a mathematical expression. Absorbed fractions for point sources were obtained by integrating the kernels over spheres. Photon dose kernels and absorbed fractions were generated for the following radionuclides: I 123, I-124, I-125, I-131, In-111, Cu-64, Cu-67, Ga-67, Ga-68, Re-186, Re-188, Sm 153, Sn-117m, Tc-99m. The Monte Carlo simulation was verified by comparing the dose kernels to published monoenergetic photon kernels. Further validation was obtained by generating an I-125 brachytherapy seed kernel and comparing it with published data. Since Monte Carlo simulation was initialized by sampling from the complete photon spectra of these radionuclides, interpolation between monoenergetic kernels and absorbed fractions was not required. The absorbed fraction due to uniform spherical distributions can be directly applied for use in internal dosimetry. In addition, the kernels can be used as input for three dimensional internal dosimetry calculations. PMID- 8724751 TI - Comment on "Improved conjugate view quantitation of I-131 by subtraction of scatter and septal penetration events with a triple energy window method" [Med. Phys. 22, 1637-1643 (1995)]. PMID- 8724752 TI - Design and optimization of an aperiodic ultrasound phased array for intracavitary prostate thermal therapies. AB - A 57 element aperiodic linear phased array was designed and constructed to investigate the feasibility of using transrectal ultrasound for the thermal therapeutic treatment of prostate cancer and benign prostatic hyperplasia. A method of reducing grating lobe levels by using optimized random distributions of unequally sized elements is introduced. Using this technique, array periodicity is avoided, making it feasible to use larger elements and hence fewer elements and amplifier channels, while still achieving acceptable power field patterns. Acoustic power field simulations determined that the grating lobe levels associated with selected aperiodic element distributions were approximately 30% 45% less than those associated with periodic element spacing and the same average element width. Or by using aperiodic rather than periodic element distributions, the average element width could be increased by approximately 20%-35% (approximately lambda/4.4), while maintaining a constant grating lobe level. Prior to construction of the 57 element array, the power capabilities of this type of array were demonstrated with a 16 element aperiodic phased array, which delivered over 28 W of acoustical power per cm of array length while focused. The power field patterns produced by the 57 element array closely matched the field patterns predicted by the theoretical model used in the simulations. The array produced acceptable power field patterns for foci at depths up to 5 cm and up to 2 cm off the center axis, in addition to producing multiple foci simultaneously. Based on the power capabilities and field patterns, this aperiodic array design has the potential to be incorporated into a clinical heating device as a means of delivering thermal therapies to the prostate and other target volumes close to body cavities. PMID- 8724754 TI - [Isolation and characterization of bacterial strains resistant to arsenical compounds]. PMID- 8724755 TI - [Scanning tunneling and electron microscopy of parasporal crystals in Bacillus thuringiensis]. AB - Comparison of the forms of parasporal inclusions in Bacillus thuringiensis subsp. kurstaki and B. thuringiensis subsp. tenebrionis under a scanning electron microscope showed that, in addition to bipyramidal and cuboid crystals, cells of the subspecies kurstaki are able to form crystals taking the shape of flat parallelopipeds typical of the subspecies tenebrionis. The inclusions in the subspecies tenebrionis vary in size and electron density. Examination of the structure of crystals in B. thuringiensis subsp. tenebrionis under a scanning tunneling microscope showed that delta-endotoxins in the crystals are assembled in globules 5.0 nm in diameter and 1.0 nm in height. The globules are connected by its bases and form chains. The subunits of tetragonal crystals in the subspecies tenebrionis are vertically oriented with respect to the chain axis, as was revealed by other methods for subunits of bipyramidal crystals of the subspecies kurstaki. PMID- 8724756 TI - [Protein engineering of artificial proteins]. PMID- 8724757 TI - [Distribution of parallel complementary sequences in the regulatory region of human mitochondrial DNA]. PMID- 8724758 TI - [Interaction of yeast TATA-binding protein with short promotor segments]. PMID- 8724759 TI - [Sequences, homologous to binding sites for eukaryotic topoisomerase I, in mycoplasma genomes]. PMID- 8724760 TI - [Site specific alkylation of dual-stranded DNA of the murine c-fos gene promotor region and modulation of transcription by derivatives of pyrimidine oligonucleotides, containing residues of 2-chlorethylamine]. PMID- 8724761 TI - [Analysis of polymorphism of two hypervariable regions of the human genome in the Russian population of Moscow using the polymerase chain reaction]. PMID- 8724762 TI - [Alternative splicing of 5'-terminal exons of the human tryptophanyl-tRNA synthetase gene]. PMID- 8724763 TI - [Heterogeneity of the 5'-untranslated area of prolactin receptor mRNA in rat liver]. PMID- 8724764 TI - [Kinetic characteristics of chimeric metalloproteinases from bacilli]. PMID- 8724765 TI - [Connection between charged clusters of gamma-crystallin with the internal protein structure]. PMID- 8724766 TI - [Interaction of RNAase H from E. coli with modified duplexes. II. Duplexes, containing oligo(1-beta-D-2'-deoxy-threo-pentofuranosyl)thymidylate]. PMID- 8724767 TI - [Orthorhombic crystals of the oligonucleotide d(GpGpCpC). Comparison with hexagonal and trigonal crystalline modifications]. PMID- 8724768 TI - [Comparative analysis of the structure and degree of stability of hydrophobic nuclei from certain globular proteins]. PMID- 8724769 TI - [Transition of "genomic size" DNA from an isotropic to a liquid crystalline state]. PMID- 8724770 TI - [Increase in binding specificity of a peptide-nucleic acid with DNA]. PMID- 8724771 TI - [Structure-activity analysis of amino acid sequences using the Internet]. PMID- 8724772 TI - [Noncanonical forms of DNA in the DARC146 sequence]. PMID- 8724773 TI - [Protein environment of matrices in the decoding region of 80S ribosomes from human placenta from data of affinity modification of mRNA analogs- oligoribonucleotide derivative]. PMID- 8724774 TI - [Resistance of hybridoma cells, producing antibodies against the binding subunit of plant toxin MLI, to the cytotoxic effect of this toxin]. PMID- 8724775 TI - [Transfection of a point pressure: a rapid and effective method of introducing macromolecules into mammalian cells]. PMID- 8724776 TI - [Computer system "AutoGene" for automatic analysis of nucleotide sequences]. PMID- 8724777 TI - [Effect of the structure of the prepromotor region on transcription using phage T7 RNA polymerase]. PMID- 8724778 TI - [VirD2-independent, but MobA-dependent transfer of plasmid DNA from a broad circle of hosts from agrobacterium into the plant cell nucleus]. PMID- 8724779 TI - [Preparation of recombinant envelope proteins from spherical phytoviruses and study of prospects for using them for immunoenzyme diagnosis]. PMID- 8724781 TI - [Errors in reproducing experimental studies of the muscle branched enzyme, amylose isomerase]. PMID- 8724780 TI - [Use of peptide-nucleic acids in the "Achilles heel" method]. PMID- 8724782 TI - [The problem of existing errors in reproducing the Petrov method of isolating amylose isomerase--a branching enzyme]. PMID- 8724783 TI - [Regenerating hematopoietic tissue as a source for a low molecular weight factor that inhibits leukemic cell proliferation]. AB - Using a successive set of chromatographic methods, a low molecular weight factor inhibiting leukemic cells proliferation has been isolated from an aqueous extract of regenerating hematopoietic tissue. This factor inhibits DNA synthesis in leukemic cells about 1000-fold more effectively than the original extract. The factor activity does not change after treatment by proteinase K; however, successive action of endo-alpha-N-acetylgalactose aminidase and proteinase K significantly decreases the efficiency of the factor action. This factor whose molecular mass is less than 3 kDa seems to be of glycopeptide origin. PMID- 8724784 TI - [Bacteriolytic enzyme preparation lysoamidase. Purification and some properties of bacteriolytic peptidase L1]. AB - The bacteriolytic peptidase L1 has been isolated from the enzyme preparation of lysoamidase capable to lyze cell walls of gram-positive bacteria using ion exchange chromatography and gel filtration. Some physico-chemical properties of the enzyme have been established. The molecular mass of L1 is 21 kDa, the pH optimum for Staphylococcus aureus cell lysis is 7-11. The optimal concentration of the buffer is 50 mM; the temperature optimum is 70 degrees C; the half inactivation temperature is 55 degrees C. PMID- 8724785 TI - [Operational stability of catalase and its conjugates with aldehyde dextrans and superoxide dismutase]. AB - Catalase, superoxide dismutase (SOD) and catalase-superoxide dismutase conjugates with aldehyde dextrans have been prepared in aqueous media and surfactant microemulsions. The catalytic activities of catalase and its conjugates were characterized by first order rate constants in successive cycles of the biocatalysts. The rate constants for catalase and its conjugates inactivation by hydrogen peroxide, kin, and the rate constants for catalase complex I interaction with H2O2, k2, were determined simultaneously from the full kinetic curves for H2O2 decomposition in 1/In[(H2O2)0/[H2O2]t)-1/t coordinates. The kin and k2 values were calculated under variable conditions of the catalase reaction and at varying concentrations of the biocatalysts and hydrogen peroxide as well as in successive cycles of the biocatalysts used for H2O2 decomposition. The utility of the kinetic parameters, kin and k2, for characterizing catalase and its conjugates inactivation and their reactivity in catalase reactions has been demonstrated. The reciprocal action of catalase and SOD on their operational stabilities in enzymatic reactions of H2O2 decomposition is discussed. Catalase conjugation to aldehyde dextrans and SOD in microemulsions enhances the stabilities of the both enzymes. PMID- 8724786 TI - [Increase in oxygen-transporting efficiency of hemoglobin when chemically modified]. AB - Polycondensation of human hemoglobin with glutaraldehyde premodified with reagents bearing acid groups (glutamic acid or sodium bisulfite), has been carried out. The resulting modified hemoglobin displays better oxygen transport characteristics in comparison with native hemoglobin. PMID- 8724787 TI - [Biochemical characteristics of aminostigmine--a new anticholinesterase agent]. AB - The properties of aminostigmine in comparison with those of other carbamate inhibitors of cholinesterases have been studied in vitro using potentiometric titration and Ellman methods. The bimolecular constants of the inhibition rate of acetyl-, butyryl- and propionylcholinesterase were found to be equal to (8.0 14.0).10(5) (3.8-7.7).10(5) and 11.0.10(5) M-1.min-1, respectively. In terms of inhibitory activity, aminostrigmine is comparable to neostigmine methylsulphate, being inferior to physostigmine and superior to pyridistigmine. The rate of decarbamylation of acetylcholinesterase inhibited by aminostigmine measured by the dilution method, by creating excessive acetylcholine and by dialysis is characterized by k2c constants equal to (1.1-1.6).10(-2), (2.5-2.8).10(-2) and 0.025.10(-2) min-1, respectively. On the whole, aminostigmine belongs to slowly reversible inhibitors. Being carbamylated by aminostigmine, the enzyme is resistant to reactivation by TMB-4 and HI-6. At (4-6).10(-7) M aminostigmine prevents by 50% the irreversible binding of cholinesterase by certain organophosphate inhibitors of cholinesterase when the latter are used at concentrations needed to inhibit the enzymatic activity by 85-90%. PMID- 8724788 TI - [Isolation of growth modulating factors secreted by human malignant melanoma cells]. AB - Novel low molecular mass growth-modulating factors-F0.5 (0.5-1.0 kDa), F1 (approximately 1 kDa) and fraction F8 (8-12 kDa)-have been isolated from a serum free culture of human malignant pigmented melanoma mS cells using ultrafiltration, affinity and low and medium pressure gel chromatography. Factor F0.5 (10(-9)-10(-7) M) had a growth-stimulating effect on melanoma mS cells exceeding by a factor of 2 and 3 that of 5% embryonic calf serum. Factor F1 and fraction F8 inhibited melanoma cell growth when used at concentrations of 10(-9) 10(-5) M and higher than 10(-6) mg/ml, respectively. The effect of fraction F8 was concentration-dependent; that of factor F1 was more complex. The inhibiting action of fraction F8 was more pronounced when amelanotic cells of human malignant melanoma BRO were used at concentrations above 10(-8) mg/ml. Neither factors F0.5 and F1 nor fraction F8 influenced the growth of human lung fibroblast Lech 240 cells. The growth-modulating factors can participate in the autocrine regulation of malignant melanoma cell growth. PMID- 8724789 TI - [Participation of DnaK chaperone and ATP in in vivo folding of firefly luciferase, synthesized by E. coli cells]. AB - The time-courses of protein accumulation and luciferase activity of Luciola mingrelica firefly luciferase synthesized on plasmid pJG lambda in E. coli strains omega 238 and B178groE7 (DnaK- and GroEL-deficient, correspondingly) and omega 237 and W3110 that are control to them, have been studied. It was shown that the amounts of the luciferase protein synthesized by all strains was approximately equal to 3.0-3.9% of the total cell protein. Luciferase was synthesized in a catalytically inactive form and transformed into an active enzyme during incubation at 21 degrees C. In the DnaK-deficient E. coli strain omega 238, the enzyme transformation into a catalytically active form did not occur which provides evidence for participation of the DnaK chaperone in transformation of newly synthesized luciferase into catalytically active form. Luciferase folding was accelerated with an increase in ATP concentration inside the cell. It is possible to increase the ATP concentration inside the cell by treatment with polymyxin and addition of ATP to the culture medium, which significantly accelerates the luciferase folding. PMID- 8724790 TI - [Monoclonal antibodies to 8-oxo-2'-deoxyguanosine (8-hydroxyguanosine). Characteristics and use for determining DNA damage by active forms of oxygen]. AB - It has presently been established that the guanine base is one of the most sensitive and biologically significant target for the damaging action of active oxygen species on DNA, 7.8-dihydroxy-8-oxoguanine (8-oxoguanine, 8 hydroxyguanine) being the major degradation product and the most essential biomarker for DNA damage by active oxygen species. Murine monoclonal antibodies (MAbs) specifically recognizing 8-oxoguanine have been raised. Using competitive solid phase immunoenzymatic assay. (IEA) with peroxidase-antiperoxidase (PAP method), a quantitative assay of this degradation product and characterization of affinity and specificity of MAbs have been carried out. The affinity constant (Kaff) Mabs for 8-oxo-2'-deoxyguanosine is equal to 1.3-10(6), that for 8-oxo guanosine-to 1.10(6), exceeding by more than three orders of magnitude the Ka values for natural guanyl nucleosides and other possible cross-structure analogs. IEA was used to determine the DNA degradation product in gamma-irradiated DNA. The radiation-chemical yield of 8-oxoguanine (G = 0.57 molecules per 100 eV) is consistent with those obtained by other methods. PMID- 8724791 TI - [Analysis of the effect of replacing Lys(-20) in the alkaline phosphatase signal peptide on secretion of this enzyme]. AB - The effect of substitutions for the positively charged Lys(-20) in the N-terminal domain of the E. coli alkaline phosphatase signal peptide on enzyme secretion has been studied. Mutant alkaline phosphatases were obtained by the amber-suppressor method. An amber mutation was introduced in the appropriate position of the alkaline phosphatase gene using oligonucleotide-directed mutagenesis. This was followed by mutant protein synthesis in E. coli strains producing amber suppressor tRNAs specific for Tyr, Gly, Ala, Glu, Phe, His, Cys, and Pro. All the mutant proteins can by translocated through the cytoplasmic membrane and form in the periplasm a molecule possessing an enzymatic activity. However, some amino acid substitutions decrease the rate of protein maturation their effect depends not only on the charge of the amino acid residue but also on its nature. Thus, introduction of positively charged. His and the polar uncharged Tyr is without effect, while negatively charged Glu and hydrophobic Ala, Phe and Pro residues as well as Gly and Cys have an inhibiting action. The results obtained testify to the importance of the signal peptide terminal domain primary structure in secretion. PMID- 8724792 TI - What are the risks of third-generation oral contraceptives? Are third-generation oral contraceptives safe? PMID- 8724793 TI - Epidemiological disasters. PMID- 8724794 TI - On the reporting of results on assisted human conception. Plea for realism and honesty in results of infertility treatment. PMID- 8724795 TI - Potential health hazards of assisted human reproduction. Possible immunological complications. PMID- 8724796 TI - Frozen embryos: too cold to touch? Frozen pre-embryos in Denmark. PMID- 8724797 TI - Decreased amounts of antibodies to 22 and 18 kDa antigens in the peritoneal fluid of patients with endometriosis. LE Group d'Investigation en Gynecologie. AB - Accumulated evidence implicates immunological alterations in endometriosis. The purpose of this study was to look for variations in antibodies to distinct antigens in peritoneal fluid of women with and without endometriosis. Peritoneal fluid was aspirated from 17 women undergoing laparoscopy for tubal ligation and 37 patients complaining of symptoms of pain and /or infertility. Peritoneal fluid antibodies to a standard preparation of peritoneal fluid antigens were detected by Western blot analysis using peroxidase-labelled anti-human immunoglobulin G antibodies specific to the Fc region. Antibodies to distinct antigens were quantified by estimating the ratio of the relative optical density between samples and a standard amount of antibodies. Marked changes were found in the antibody detection to two antigens having apparent molecular weights of 22 and 18 kDa. The intensity of the antibody signal was significantly weaker in the peritoneal fluid from endometriosis patients (0.36 +/- 0.06 and 0.46 +/- 0.06) compared with that in women without endometriosis (0.62 +/- 0.08 and 0.75 +/- 0.06). It was also weaker in patients without endometriosis presenting with infertility (0.36 +/- 0.07 and 0.47 +/- 0.08), but only the 18 kDa antigen result was significant. After adjusting for infertility, the P values for the 18 and 22 kDa bands were 0.03 and 0.28 (not significant) respectively in the group of endometriosis patients. These changes were not related to the phase of the menstrual cycle. These data suggest an alteration in the immune response to two distinct antigens in the peritoneal fluid from women with endometriosis and infertility. Further evaluation of these two antigens and their antibodies would be of interest to help understand endometriosis and its associated infertility. PMID- 8724798 TI - Calcitonin gene-related peptide reverses the hypertension and significantly decreases the fetal mortality in pre-eclampsia rats induced by N(G)-nitro-L arginine methyl ester. AB - We recently established that the chronic inhibition of nitric oxide production with N(G)-nitro-L-arginine methyl ester (L-NAME) increases blood pressure and fetal mortality in pregnant rats. Using this animal model, we have investigated whether calcitonin gene-related peptide (CGRP) can reverse the pre-eclampsia-like conditions produced by L-NAME. CGRP and L-NAME were chronically infused s.c. into pregnant rats separately or together starting on day 17 of gestation; a control group was given saline infusions. Systolic blood pressure was measured on gestational days 17, 18, 19 and 22 and post-partum days 1 and 2. The weight and mortality of the pups were recorded immediately after spontaneous delivery. Animals treated with L-NAME exhibited significant elevations of blood pressure on days 18, 19 and 22 of gestation and during post-partum, increased pup mortality (18.4 versus 0.0%) and decreased pup weights (5.14 +/- 0.07 versus 6.20 +/- 0.06 g). The co-administration of L-NAME and CGRP prevented the gestational (not the post-partum) L-NAME hypertension and decreased pup mortality to 6.4% but did not reverse the decreased fetal weight (5.31 +/- 0.06 g). Our data indicate the CGRP (i) participates in regulation of the vascular adaptations that occur during normal pregnancy, (ii) has beneficial effects on the hypertension and increased mortality of experimental preeclampsia, and (iii) may exert differential effects on the systemic (i.e. maternal) and fetal components of utero-placental circulation. These findings may have important clinical implications. PMID- 8724799 TI - An HPLC radiotracer method for assessing the ability of L-cysteine prodrugs to maintain glutathione levels in the cultured rat lens. AB - PURPOSE: To apply a high performance liquid chromatographic radiotracer method to test a variety of L-cysteine prodrugs and one dipeptide prodrug for their ability to synthesize glutathione in cultured rat lenses. METHODS: Rat lenses were incubated for 48 h in a medium containing [14C(U)]-glycine and prodrugs. Following homogenization and derivatization, lens extracts were analyzed to determine the extent of biosynthetic incorporation of this labeled amino acid into [14C]-glutathione using high performance liquid chromatography with radioisotope and ultraviolet absorption detection. All of the thiazolidine prodrugs contained masked sulfhydryl groups to stabilize them against air oxidation. L-buthionine-(S,R)-sulfoximine-an inhibitor of the first step in glutathione biosynthesis-was present in media containing the dipeptide prodrug. RESULTS: In all cases, a large [14C]-labeled peak eluted just prior to [14C] glutathione. This peak had some characteristics of the mixed disulfide of glutathione and L-cysteine, viz., L-cysteine/glutathione disulfide, but requires further investigation in order to be positively identified. Of the eleven L cysteine prodrugs investigated, the most effective was 2(R,S)-methylthiazolidine 4(R)-carboxylic acid, which increased the rate of [14C]-glutathione biosynthesis 35% over that of the controls. A number of other L-cysteine prodrugs were somewhat effective, increasing glutathione synthesis 5-30% over the controls, while several L-cysteine prodrugs were totally ineffective. The only dipeptide prodrug investigated, viz., gamma-L-glutamyl-L-cysteine ethyl ester, increased the biosynthesis of [14C]-glutathione 18% over control. Biosynthetic rates based on ultraviolet absorption of the derivatized glutathione demonstrated a similar pattern, the compounds most effective in synthesizing [14C]-glutathione generally yielding the highest ultraviolet glutathione concentrations and the ineffective compounds showing the lowest concentrations. CONCLUSIONS: 2(R,S) methylthiazolidine-4(R)-carboxylic acid, 2(R,S)-n-propylthiazolidine-4(R) carboxylic acid and N-acetyl-L-cysteine were the only compounds that were statistically significant in yielding higher levels of both ultraviolet and radioactive glutathione as compared to their respective controls. Thus, these prodrugs have very promising anti-cataract potential. PMID- 8724800 TI - Preventive strategies for trachoma control. PMID- 8724801 TI - [In situ hybridization of cells infected by Chlamydia trachomatis]. AB - Hybridization in situ of chlamydia trachomatis allows identification of the bacteria. Moreover exact chlamydia trachomatis localization at tissue level can be demonstrated. It seems to us very important to know for instance in conjunctiva were chlamydia trachomatis are located. We used DNA DIGOXIGENIN labelled plasmidic probe of 503 b.p. Detection of chlamydia trachomatis on paraffin embedded cells becomes possible. Specificity is 100% as plasmidic sequence is not found in other bacteria. Sensitivity seems to be good quality on experimental model. At present time, it is under clinical evaluation. This method could complete PCR for chlamydia detection in case of controversial diagnosis. PMID- 8724803 TI - Developments for a global approach to trachoma control. AB - Because of the life-style/environment-related determinants of the disease, trachoma control activities should be multisectorial and multidisciplinary in order to be effective and long lasting. Medical and epidemiological components to be included in the global approach have largely benefited by the existence of the simplified trachoma grading system. In other respects, long-term outcomes of trichiasis surgery are better evaluated and various surgical procedures are available. On the other hand, despite the knowledge that community participation is one of the essentials of trachoma control activities, this element all too often remains neglected. Therefore, realistic and practical community approaches need to be specified which are suited to various socio-cultural settings. Thanks to the support of the Edna McConnell Clark Foundation (USA), the World Health Organization's Programme for the Prevention of Blindness has prepared a set of documents, addressing the different aspects of trachoma control activities. PMID- 8724804 TI - The ophthalmologist's role in Chlamydia trachomatis infection. AB - Chlamydia Trachomatis (C.T.) most commonly causes cervicitis in women, often without overt symptoms; when untreated, it produces a clinically inapparent but persistent mild infection or an evident salpingitis. Either processes may lead to scarring of the falloppian tubes and fertility problems. A group of 30 women 20 35 years aged has undergone an ophthalmological routine examination; 7 of them presented a chronical follicular conjunctivitis and history of yellow-white cervical discharge, while the remaining 23 had a mild chronical follicular conjunctivitis but not genital symptoms. Each patient has undergone an immunofluorescent test with monoclonal antibody specific for C.T. on the samples collected by genital and conjunctival scraping and cytological evaluation of the above samples with Giemsa staining. 6 out of 7 patients with genital discharge were positive for C.T., one of which even for eye samples too; all eyes samples showed lymphomonocytes and neutrofilic cells in their eye samples. An additional 3 cases negative for C.T. showed this kind of cells in the eye scraping. The preliminary results of this study on mild follicular chronical conjunctivitis suggest that the Ophthalmologist could play an important role in order to make an early diagnosis in genital and eye infection by Chlamydial Trachomatis. PMID- 8724805 TI - [Effect of water supply on the incidence of trachoma: preliminary results of a prospective study in Mali]. AB - This prospective study (January 1993-January 1994) was carried out in two groups of villages of the district of Ouelessebougou (Mali). The objective of the study was to assess the impact of installing water supply boreholes on the incidence of trachoma. A prospective group of 4 villages, where boreholes were sunk at the beginning of the study, was compared to a control group of 4 villages where there are no boreholes. The examination concerned children below the age of 10 years (N = 700) and was carried out using binocular loupes, according to the simplified procedure for trachoma grading proposed by the WHO. Our results show that in the prospective group, the annual rate of incidence of inflammatory trachoma was 8% (cl: 5.8-10.7) while in the control group it was 14% (cl: 9.6-10.9). RR: 0.58 (0.37-0.90). Likewise the rate of spontaneous healing was 12% (Cl: 9.1-14.9) in the prospective group and 5% (cl: 2.4-9) in the control group. Relative Risk was 2.60 (1.36-4.9). These statistically significant results show that the risk of contracting trachoma is reduced by half in the villages where a borehole has been sunk. PMID- 8724806 TI - [Epidemiologic aspects of retinoblastoma in a tropical region]. AB - The authors are publishing the epidemiologic aspects of retinoblastoma in a cameroonian area. The study is prospective and is carried out in the eye clinic of CHU over a period of 7(1/2) years. We include all patients diagnosed and confirmed histologically after enucleations to have retinoblastoma. The frequency of retinoblastoma is 0.83% per 10000 consultations. The sex ratio is 1/6 (two boys and 12 girls) ans the mean age of patients on first consultation (visit) is 38 months (between 3 months and 6 years 7 months). 85.55% of the retinoblastomas are sporadic and the disease is lethal in 78.57% of cases. PMID- 8724807 TI - [Ocular cysticercosis (O.C.) in Madagascar (apropos of 6 cases)]. AB - Cysticercosis is due to cysticercus cellulosae and was know since a long time. Human is an intermediate occasional host by ingesting accidentally eggs of tenia. In Madagascar, neurocysticercosis are predominating (55%), occular localisations are not frequent (5%) and concerned particularly vitreous humour and retina. Diagnosis relies on serology: Elisa test, then Western Blot completed with research of circulating antibody and antigen in the acqueous humor by immocapture. Anatomopathologic test permits to confirm the diagnosis. Contribution of tomodensitometry is not conclusive therapeutic is summed up to a specific and symptomatic treatment with surgical extirpation when out means allow it. PMID- 8724808 TI - [Acute epidemic hemorrhagic conjunctivitis (AEHC) in Madagascar 1991]. AB - An epidemic of A.E.H.C. was broken out everywhere in Madagascar during hot season from September 1990 to May 1991 with an important acuteness in February and March. Clinical symptoms are those of A.E.H.C. to enterovirus with some particularities. Virological study shows cytopathogenic effect of enterovirus and microbiology shows the existence of several bacterial germs particularly staphylococcus epidermidis. The discovery of two cases of streptococcus pneumonia and the absence of chlamydia trachomatis make the originality of our cases. The efficacy of the association antibiotic and steroids is spectacular. PMID- 8724809 TI - [[Trabeculectomy in Madagascar. Retrospective study over 3 years]. AB - This study reports the results of the evolution of ocular pressure, glaucoma excavation and complications after trabeculectomy particularly cataracts observed in 47 cases of chronic glaucoma. Our chronic glaucomas are frequent in male with an average of age of 60,5 years. Normalisation of ocular tension has been obtained in 87.23% of cases. However in 10.53% of cases, an adjuvant medical treatment has been necessary. Ocular pressure figure raised up progressively after trabeculectomy to reach some limit from which it remains stable. The best ocular pressure control has been observed first in glaucoma with opened angle, then in post traumatic glaucoma. Visual acuteness was improved in 20.27% of cases. Trabeculectomy has involved a relief, a disparition of pain in all the cases. The main late complication observed is opacification of eye lens in patients aged more than 70 years old. PMID- 8724810 TI - Emerging resistance to antimicrobial agents in gram-positive bacteria. Pneumococci. AB - The prevalence of penicillin-resistant pneumococci is increasing worldwide. Methods for susceptibility testing, as well as in vitro susceptibility of penicillin-susceptible and -resistant strains to new and existing agents (including oral and parenteral streptogramins), are described. For all specimens except CSF, oxacillin screening followed by determination of penicillin minimum inhibitory concentrations (MICs) is satisfactory. For CSF, simultaneous testing of penicillin and cefotaxime or ceftriaxone by E-test is necessary. Of all available oral beta-lactams, amoxicillin yields the lowest MICs against penicillin-susceptible and -resistant pneumococci, and is the drug of choice for the treatment of otitis media. Cefotaxime and ceftriaxone yield MICs that are low enough to permit therapy of meningitic and nonmeningitic infections (the former in combination with vancomycin). The higher the strain's benzylpenicillin (penicillin G) MIC, the more likely it is that simultaneous resistance to nonrelated compounds such as tetracyclines, macrolides, and cotrimoxazole (trimethoprim/ sulfamethoxazole) will occur. None of the available quinolones should be used for therapy of pneumococcal infections. Of new and experimental drugs, some of the new quinolones, trovafloxacin, and oral and parenteral streptogramins are promising agents. Imipenem is epileptogenic, but meropenem has potential in the therapy of meningitis. Problematical infections caused by penicillin-resistant pneumococci include meningitis and otitis media. The optimal therapy of the latter two diseases has not yet been clearly delineated. PMID- 8724811 TI - Emerging resistance to antimicrobial agents in gram-positive bacteria. Enterococci, staphylococci and nonpneumococcal streptococci. AB - Staphylococci (Staphylococcus aureus and coagulase-negative Staphylococcus species) and enterococci are the aetiological organisms in 47 to 52% of nosocomial blood stream infections and approximately 30% of all nosocomial infections in the US. In European intensive care units, almost half of all infections are attributed to staphylococci. The streptococci have also become increasingly important because of the modified virulence of Streptococcus pyogenes strains, and the emerging role of the viridans group streptococci as a cause of potentially fatal bacteraemia in the neutropenic host. Resistance to available antimicrobial agents is increasing and includes, in particular, resistance to the glycopeptides (vancomycin and teicoplanin) amongst enterococci, resistance to penicillinase-resistant penicillins (oxacillin and methicillin) and fluoroquinolones (ciprofloxacin and ofloxacin) amongst staphylococci, and resistance to penicillin and some other beta-lactams amongst viridans group streptococci. New compounds for effective therapy of infection with antimicrobial resistant Gram-positive species are needed urgently. To this end, the streptogramin combinations [quinupristin/dalfopristin (RP 59500; Synercid)], everninomycin derivatives (SCH 27899), oxazolidinones (U-100572, U-100766) and several newer fluoroquinolones (clinafloxacin, DU 6859a, grepafloxacin, levofloxacin, sparfloxacin, trovafloxacin) are under rapid development and clinical investigation. PMID- 8724812 TI - Streptogramins. A unique class of antibiotics. AB - Streptogramin antibiotics represent a unique class of antibacterials in that each member of the class consists of at least 2 structurally unrelated molecules: group A streptogramins (macrolactones) and group B streptogramins (cyclic hexadepsipeptides). Both group A and group B streptogramins inhibit protein synthesis at the ribosomal level, and they act synergistically against many isolates, their combination generating bactericidal activities and reducing the possibility of emergence of resistant strains. The mechanisms of acquired resistance to group B streptogramins are similar to those induced by erythromycin, but group A streptogramins remain unaffected by target modifications and active efflux. The pharmacokinetic parameters of group A and group B streptogramins in blood are quite similar. In addition, both the A and B groups penetrate and accumulate in macrophages and in the bacterial vegetations of experimental endocarditis. There are important structural and biological differences between the streptogramins and the macrolides. The main differentiating features are the rapid anti-bacterial killing of streptogramins and the rarity of cross-resistance between the 2 groups of antibiotics. PMID- 8724813 TI - Mechanism of action of streptogramins and macrolides. AB - Protein synthesis is catalysed by ribosomes and cytoplasmic factors. Bacterial ribosomes (70S) are made up of 2 subunits (50S and 30S) containing ribosomal RNA (rRNA) and ribosomal proteins: the 30S binds messenger RNA and begins the ribosomal cycle (initiation), whereas 50S binds transfer RNA (tRNA) derivatives and controls elongation. The key reaction, peptide bond formation, is promoted by the catalytic centre of 50S (the peptidyl transferase centre), and the growing peptide chain (peptidyl-tRNA) attached at the donor P site undergoes peptide linkage with an aminoacyl-tRNA at the acceptor A site. This reaction is inhibited by several antibiotics, the best known being chloramphenicol, and the macrolide lincosamide-streptogramin (MLS) group. These inhibitors have a reversible action, except for streptogramins that are composed of A and B components, which are bacteriostatic alone, but bactericidal when combined. The peptidyl transferase centre has been identified at the 50S surface, and the binding sites of inhibitors have been mapped within this domain: some of these sites overlap (e.g. those of macrolides, and type B streptogramins, which compete for binding to ribosomes). Chloramphenicol blocks the catalytic portion, and A streptogramins the substrate sites of the peptidyl transferase centre. Macrolides and type B streptogramins interfere with the formation of long polypeptides and cause a premature detachment of incomplete peptide chains. The synergism between types A and B streptogramins is due to induction by type A streptogramins of an increased ribosome affinity for type B streptogramins. Microbial resistance to antibiotics mainly involves inactivation of inhibitors and modification of targets (mutations of ribosomal proteins or rRNA genes). Alterations of rRNA bases can induce resistance to a single inhibitor or to a group of antibiotics (e.g. MLSB). The impact of resistance in chemotherapy is less important for streptogramins than for other inhibitors, because the synergistic effect of A and B streptogramins also applies to strains resistant to the MLSB group. It is proposed that mutations and modifications of rRNA bases induce conformational ribosomal changes that prevent antibiotics binding to the target. Conformational changes are also triggered by type A streptogramins: they are responsible for their synergism with type B streptogramins. PMID- 8724814 TI - Antibacterial activity of quinupristin/dalfopristin. Rationale for clinical use. AB - Most Gram-positive organisms are highly susceptible to the streptogramin, quinupristin/dalfopristin (RP 59500; Synercid). Minimum inhibitory concentrations for 90% of isolates (MIC90) were < or = 1 mg/L for Staphylococcus aureus, S. epidermidis, S. haemolyticus, Streptococcus pneumoniae, S. pyogenes and Listeria monocytogenes. Importantly, quinupristin/dalfopristin shows similar activity against methicillin-susceptible and -resistant strains of S. aureus, and streptococci with benzylpenicillin (penicillin G)- or erythromycin-acquired resistance. Enterococci have varying susceptibility to quinupristin /dalfopristin, although most isolates tested are susceptible to the drug, including vancomycin-resistant and multiresistant Enterococcus faecium. E. faecalis are generally the least susceptible. Among the Gram-negative respiratory pathogens Moraxella catarrhalis is susceptible and Haemophilus influenzae is moderately susceptible to quinupristin/ dalfopristin; however, Enterobacteriaceae, Pseudomonas aeruginosa and Acinetobacter spp. are resistant. The drug is active against anaerobic organisms tested, including Clostridium perfringens, Lactobacillus spp., Bacteroides fragilis and Peptostreptococcus. Synergy has been demonstrated in vancomycin-resistant and multiresistant E. faecium, and methicillin-sensitive and -resistant S. aureus with the combination of vancomycin and quinupristin/ dalfopristin. Quinupristin/dalfopristin shows antibacterial activity in vivo in animal models of infection, including methicillin-sensitive and -resistant S. aureus infection in rabbits, S. aureus and S. pneumoniae in mice, and erythromycin-sensitive and -resistant viridans group streptococci infections in rats. The drug is rapidly bactericidal against Gram-positive organisms (with the exception of enterococci) at concentrations similar to or within 4-fold of the MIC, and it has a long postantibiotic effect both in vitro and in vivo. PMID- 8724815 TI - Future prospects and therapeutic potential of streptogramins. AB - Dramatic changes in the epidemiology and susceptibility patterns of Gram-positive cocci during the last decade have mandated new approaches to the management of many bacterial infections. For example, there has been a sharp increase in the incidence of infections caused by Staphylococcus aureus, particularly those resistant to methicillin (MRSA), and methicillin-resistant coagulase-negative staphylococci, particularly those associated with foreign bodies and indwelling medical devices. Additionally, the worldwide spread of Streptococcus pneumoniae strains resistant to penicillin and macrolides, and the emergence of enterococci (particularly Enterococcus faecium) resistant to vancomycin, teicoplanin and other antibiotics, present further therapeutic problems. New antibacterial agents are urgently required to meet the challenges posed by these epidemiological trends. The semisynthetic streptogramins, a unique class of antibacterials currently under development, offer promise in the treatment of such multiresistant infections. Possible future applications include treatment of infections caused by the following organisms: MRSA, enterococci resistant to vancomycin, macrolides or lincosamides; and beta-lactam-resistant streptococci. They may also prove useful as therapy for children with staphylococcal infection and patients with multiresistant infections who are unable to tolerate vancomycin, including patients with skin and soft tissue infections caused by Gram-positive pathogens, patients with osteomyelitis, foreign body associated infections, endocarditis and sepsis due to Gram-positive bacteria. Clinical trials are required to evaluate the efficacy and tolerability of streptogramins in these settings. PMID- 8724816 TI - Atlas of cancer mortality in central Europe. PMID- 8724817 TI - Carl Arthur Goresky. August 25, 1932 to March 21, 1996. PMID- 8724818 TI - Insulin-degrading enzyme. AB - The authors review recent research on an enzyme hypothesized to play a major role in the degradation of insulin. After binding to its receptor on the cell surface, insulin is internalized by receptor-mediated endocytosis and degraded within components of the endosomal apparatus. Degradation of insulin is important in the termination of signaling and clearance of the circulating hormone. It has been proposed that insulin-degrading enzyme (IDE), an evolutionarily conserved, neutral thiol-metalloendopeptidase, plays a crucial role in the degradation of internalized insulin in many types of cells. Despite the substantial evidence supporting the importance of IDE in cellular insulin degradation, there is controversy over its mode and site of action, mainly because of its cytosolic location. Its physiological location in cells has recently been elucidated through subcellular fractionation of liver parenchyma and through immunofluorescence microscopy of stably transfected Chinese hamster ovary cells that overexpress IDE. These experiments have excluded the presence of the enzyme in endosomes and have defined a peroxisomal location, consistent with the presence of a peroxisomal targeting sequence at the carboxyl terminus of the protein. Recently, researchers have demonstrated the functional significance of peroxisome-associated IDE (type I peroxisomal enzyme) in degrading cleaved leader peptides of peroxisomal proteins targeted by the type II motif. IDE is the first cloned and characterized proteinase to be localized to peroxisomes. Moreover, IDE appears to be a member of a newly identified superfamily of metalloendopeptidases that has an HXXEH active-site motif. Although fundamental questions concerning the biological role of IDE remain, its high degree of evolutionary conservation suggests that it must have important functions and multifaceted biological significance. PMID- 8724819 TI - The role of lipids in cardiovascular disease: lessons from rare mutations and special populations. AB - OBJECTIVE: To review approaches to determining the genetic basis of atherogenesis, the interaction of genes with the environment and with other genes, and the contribution to knowledge of data from subjects with rare mutations and from genetically isolated populations. DATA SOURCES AND SELECTION: Current literature, including studies of Canadian families with genetic mutations by the author, on the role of genetic factors in dyslipidemia and atherosclerosis. DATA EXTRACTION: Studies that have shown effects of gene products on plasma levels of cholesterol and lipids and subsequent atherogenesis. DATA SYNTHESIS: Studies in families with rare mutations indicate that a single genetic change has a profound effect on phenotype, but the possible clinical application of this information is restricted mainly to members of the extended family. In contrast, studies carried out in large populations show that many genetic factors each have a small impact on phenotype and that these effects are further obscured by environmental factors. The clinical relevance of such findings is unclear. CONCLUSIONS: Ideally, the information gained from new molecular approaches to identifying the determinants of the intermediate phenotypes of atherosclerosis should be integrated into evidence-based practice. However, the complicating factors are formidable. The use of genetic information to identify subjects at high risk of the clinical end points of atherosclerosis and to enhance health care delivery to such subjects is questionable. PMID- 8724820 TI - Molecular genetic aspects of hyperhomocysteinemia and its relation to folic acid. AB - In a review of research by the author and her colleagues, the genetic basis of hyperhomocysteinemia and the relation between this condition and plasma folate levels are elucidated. There has recently been renewed interest in homocysteine metabolism because hyperhomocysteinemia has been associated with occlusive arterial disease and neural tube defects. The article focuses on a critical enzyme of folate metabolism, 5,10-methylenetetrahydrofolate reductase. A deficiency of this enzyme results in hyperhomocysteinemia and a wide variety of neurologic and vascular symptoms. Molecular genetic analysis of the enzyme has led to the identification of nine rare mutations associated with a severe deficiency phenotype as well as one common mutation (found in 35% to 40% of alleles in the general population) that is proposed as a risk factor in some forms of cardiovascular disease and in neural tube defects. PMID- 8724821 TI - What difference does it make to be treated in a clinical trial? A pilot study. AB - OBJECTIVE: Pilot study to characterize treatment differences between patients treated in clinical trials and those treated in a clinical setting. Previous studies have shown higher survival rates for participants in trials of cancer therapy. This difference is observed even after rates are adjusted for important covariates such as age and stage of disease. DESIGN: Retrospective chart review. SETTING: Oncology outpatient department in a tertiary care hospital. PATIENTS: Ninety women 18 to 70 years of age with early-stage breast cancer who were diagnosed in 1990. Fifty-one of the women were treated through clinical trials and 39 were treated outside of clinical trials. OUTCOME MEASURES: Number of blood tests, telephone calls, clinic visits and imaging procedures as well as intensity of chemotherapy and use of radiation therapy. The age of the patient and the stage of disease were important covariates. RESULTS: After the analysis was controlled for patient age and stage of disease, patients treated through a clinical trial were more likely to receive standard-dose chemotherapy (p = 0.020, 95% confidence interval 1.20 to 200.73) and more frequent blood tests (p < 0.001, 95% confidence interval 1.02 to 1.13) than other patients treated in the clinic. CONCLUSIONS: Our results provide a plausible mechanism for the observed survival advantage for participants in clinical trials in oncology. Further study is called for. If these results are confirmed, they have important implications for informed consent to participate in clinical trials and for clinical practice. PMID- 8724823 TI - Folic acid and prevention of neural tube defects: a study of Canadian mothers of infants with spina bifida. AB - OBJECTIVES: To determine the diffusion of information about preventing neural tube defects (NTDs) through folic acid consumption by examining whether mothers of Canadian children born with spina bifida, who had become pregnant at least a year after evidence of the preventive effect of folic acid had been published, had taken sufficient amounts of folic acid in the periconceptional period and were aware of this important new information. DESIGN: Validated food-frequency questionnaire to assess folate intake. SETTING: The Hospital for Sick Children in Toronto between Jan. 4 and Aug. 16, 1994. PARTICIPANTS: Thirty mothers whose infants were being treated for spina bifida. MAIN OUTCOME MEASURES: The mothers' mean folate intake and knowledge about the protective effect of folic acid; demographic and health information. RESULTS: The mothers' mean folate intake was 0.182 mg/d (standard deviation 0.076 mg/d, range 0.02 to 0.53 mg/d), less than half the protective dose. Only 4 (13%) of the mothers had been aware of the relation between nutritional folate and NTDs when they conceived, but even they did not supplement their diets with sufficient folic acid. The medical data showed that, in addition to the failure of primary prevention of NTDs, secondary prevention through diagnostic tests during pregnancy were also inadequate. CONCLUSIONS: Our study, one of the first to be conducted after the role of folate in preventing NTDs was confirmed, reveals that, in one of the most advanced countries in the world, this new information has had no effect on patients' folate intake. Unless food is fortified with folate, the estimated 400 to 800 annual cases of NTDs in Canada will not be prevented. PMID- 8724822 TI - The effect of varying sodium intake on blood volume, forearm blood flow and vascular responsiveness to sympathetic stimulation in pre-ascitic cirrhosis. AB - OBJECTIVES: To assess, in patients with well-compensated pre-ascitic cirrhosis, (1) the extent of vasodilatation, if any, in the forearm circulation and (2) the effect of sodium status on its response to reflex sympathetic stimulation. DESIGN: Case-control study. SETTING: Clinical investigation unit of the Toronto Hospital, a tertiary referral hospital. PATIENTS: Eight male, alcoholic patients with pre-ascitic cirrhosis and 10 age- and sex-matched controls. INTERVENTIONS: Patients and controls were given a diet containing 20 mmol of sodium per day for 7 days, then a diet containing 200 mmol of sodium per day for the subsequent 7 days. On the seventh day of each diet, systemic hemodynamics, forearm circulation and effective arterial blood volume were assessed. A cold pressor test was performed after both diets to assess the response of the forearm circulation to reflex sympathetic stimulation. OUTCOME MEASURES: Heart rate, mean arterial pressure, forearm blood flow, forearm vascular resistance, central venous pressure, atrial natriuretic factor concentrations and neurohumoral pressor levels (plasma renin activity, aldosterone and plasma norepinephrine levels). RESULTS: No forearm vasodilatation was evident in the patients with pre-ascitic cirrhosis; their forearm blood flow and forearm vascular resistance were similar to those of the controls. Sodium loading did not influence baseline forearm blood flow. Cold pressor stimulus resulted in a significant decrease in forearm blood flow and a significant increase in forearm vascular resistance, mean arterial pressure, heart rate and plasma norepinephrine levels in both groups after the low-sodium diet. High sodium intake resulted in significantly greater reduction in forearm blood flow (-19%, standard error of the mean [SEM] 3% v. -8%, SEM 3%; p < 0.05) and significantly greater increase in forearm vascular resistance (+46%, SEM 7% v. +25%, SEM 8%; p < 0.05) in the patients with cirrhosis than in the controls. CONCLUSIONS: Well-compensated pre-ascitic cirrhotic patients do not have forearm vasodilatation. Sodium loading does not increase baseline forearm blood flow in these patients, but it does lead to a heightened response to reflex sympathetic stimulation. Sodium loading, with the associated sympathetic hyper responsiveness, may therefore contribute to further sodium retention in these patients. PMID- 8724824 TI - Folic acid to prevent neural tube defects: time for food fortification. PMID- 8724825 TI - Research in medical schools: rationale, priorities, roles and balance. AB - Although medical schools traditionally rest on the "three-legged stool" of research, education and service, it may often seem to the outsider that research is ascendant. In the past 50 years, medical schools' research success has been abundant; they are often most secure in contemplating their indispensable role in this domain. Recently, however, growing criticism of educational programs and increased competition for service responsibility (and the attendant revenue) from the nonacademic and private sectors have fuelled asymmetry. Research may well be the best bulwark against diminished importance or mediocrity, but it should be fortified by a new balance in which the medical schools' mission in education and service is reinforced. Unipolar concentration on the understanding of disease mechanisms must be eschewed in favour of a blended program of basic, clinical and population health sciences. Medical schools must pay greater attention to their responsibilities for training graduate students in a variety of health-related disciplines; in the future, nonphysician health care professionals will increasingly share the scientific preparation and views of physicians as they work in multiprofessional teams. Research will continue to thrive in the medical school of the future, but success will come from a careful assessment of current realities and a strategic resetting of priorities. PMID- 8724826 TI - T cell subsets in infectious and autoimmune diseases. Introduction. PMID- 8724827 TI - Differentiation and tolerance of CD4+ T lymphocytes. AB - The development of effector and memory populations of T lymphocytes is determined by antigen-induced growth and differentiation of naive T cells, and it is regulated by antigen-induced functional tolerance and cell death. CD4+ helper T lymphocytes that vary in their profiles of cytokine production and in effector functions also show distinct responses to antigens and co-stimulatory signals, and they differ in their sensitivity to tolerance induction. Thus, stimuli that trigger T cell growth and differentiation, as well as mechanisms that inhibit T cell expansion, determine both the magnitude and the nature of T cell-dependent immune responses to protein antigens. PMID- 8724828 TI - Reversal of polarized T helper 1 and T helper 2 cell populations in murine leishmaniasis. AB - T helper 1 (Th1) and Th2 cells are the major subsets of fully differentiated CD4+ T cells in the mouse. The spectrum of cytokines characteristic of each subset determines the distinctive regulatory and effector functions mediated by each subset. We have used the murine model of Leishmania major infection to study the question of whether highly polarized populations of normal T cells are as stable in their cytokine phenotype as Th clones or whether the phenotype can be altered with regulatory cytokines. Interleukin 4 (IL-4) appears to be a key cytokine for Th2 responses as it is necessary for both the initial differentiation of Th responses to L. major and the stability of ongoing responses. Furthermore, IL-4 is capable of converting highly polarized Th1 responses to Th2 responses either in vitro or when adoptively transferred to severe combined immunodeficiency mice. PMID- 8724829 TI - Differentiation of subsets of CD4+ and CD8+ T cells. AB - Our knowledge of the cytokine secretion patterns of T cells and other cells is clearly becoming more complex. The T helper 1 (Th1) and Th2 patterns may represent the extremes of a spectrum of cytokine regulatory patterns controlled by several cell types. CD8+ T cells can also secrete either Th1-like or Th2-like cytokine patterns, and they can contribute to bystander B cell activation. Interactions occur between immune cytokine regulatory networks and other systems, and pregnancy and responses against infection can profoundly influence each other. PMID- 8724830 TI - Role for CD30 antigen in human T helper 2-type responses. AB - Human T helper 1 (Th1) cells develop preferentially during infections by intracellular parasites and trigger phagocyte-mediated host defence. In contrast, human Th2 cells are responsible for phagocyte-independent host response, and they predominate during helminthic infestations and in atopic humans in response to common environmental antigens. Polarized human Th1 and Th2 cell responses play different roles in protection, and they can promote different immunopathological reactions. Strong and persistent Th1 responses seem to be involved in organ specific autoimmunity, contact dermatitis and some chronic non-allergic inflammatory disorders. Polarized Th2 responses favour reduced protection against the majority of infections, including HIV, and they are responsible for triggering allergic disorders in genetically predisposed hosts. Th1 and Th2 cells probably exhibit distinct surface markers; for example, Th2 cells express preferentially membrane CD30 and release the soluble form of CD30, which is a member of the tumour necrosis factor receptor superfamily. CD30-mediated signalling promotes the in vitro development of Th2-like cells. The expression of CD30 in HIV-infected T cells results in enhanced HIV replication, suggesting the existence of complex links among CD30 expression, production of Th2-type cytokines and immunopathogenesis of HIV infection. PMID- 8724831 TI - Regulation of T lymphocyte subsets. AB - Patterns of cytokine secretion and functional differences distinguish T lymphocyte subsets. T lymphocyte subsets are also regulated differentially. Most established CD8+ lymphocyte clones secrete gamma-interferon (IFN-gamma) but not interleukin 2 (IL-2) or IL-4. Using murine T cells which express a transgenic, antigen-specific alpha/beta T cell receptor (TCR) specific for L(d) class I major histocompatibility complex antigen, we have found that CD8+ lymphocytes can be divided into functional subsets. Freshly isolated CD8+ T cells are not cytolytic, do not proliferate and do not proliferate and do not secrete cytokines. Stimulation of TCR alone does not induce cytokine secretion, but cells become responsive to exogenous IL-2 or IL-4. Stimulation of CD28 together with TCR induces secretion of IL-2 and IFN-gamma, and cells proliferate without exogenous cytokines. Proliferation is necessary for the development of cytolytic activity. If IL-4 is present during initial stimulation, IL-4 is secreted following restimulation. Upon stimulation, some IL-4-producing murine CD8+ T cell clones express CD40 ligand (CD40L), and they potentiate proliferation and immunoglobulin secretion by small resting B cells. Thus, the CD8+ T cell subsets T cytotoxic 1 (Tc1) and Tc2 are analogous to CD4+ T helper 1 (Th1) and Th2. IL-2 production by naive CD8+ cells requires co-stimulation. IL-4 production by CD8+ T cells requires the presence of IL-4 during initial stimulation. Some IL-4-producing CD8+ T cells express CD40L following TCR stimulation and provide help for B cells. PMID- 8724832 TI - Induction and regulation of host cell-mediated immunity by Toxoplasma gondii. AB - Toxoplasma gondii is a highly infectious intracellular parasite which, if left unchecked by the immune system, rapidly overwhelms its intermediate hosts, as illustrated by the pathogenesis of toxoplasmic encephalitis in patients with AIDS. In order to insure both its host's and consequently its own survival simultaneously, T. gondii induces a potent gamma-interferon (IFN-gamma)-dependent cell-mediated immunity early in infection that controls the replication of the protozoan and facilitates transformation into the dormant cyst stage. The protective IFN-gamma is derived from three sources: natural killer cells; and CD4+ and CD8+ T lymphocytes, which can partially compensate for each other in knockout mice lacking the appropriate major histocompatibility complex restricting elements. At least two properties of the parasite appear to be responsible for the early induction of these effector cells. The first is a hydrophobic molecule (or group of related molecules) that triggers interleukin 12 (IL-12), tumour necrosis factor alpha and IL-1beta synthesis in macrophages. This response can also promote HIV replication in the same cells. The second is a superantigen activity that drives IFN-gamma-producing Vbeta5+ CD8+ T cells. These potentially lethal responses are later regulated through the triggering of IL-10 and by the induction of anergy in the superantigen-stimulated Vbeta5+ T cell population. PMID- 8724833 TI - The development of effector T cell subsets in murine Leishmania major infection. AB - Leishmania major infection has proven an exceptional model for CD4+ subset development in inbred mice. Most strains contain infection coincident with the appearance of T helper 1 (Th1) cells that produce gamma-interferon (IFN-gamma) required for macrophage activation. In contrast, mice on the BALB background are unable to control infection due to the development of Th2 cells that produce counter-regulatory cytokines, particularly interleukin 4 (IL-4), capable of abrogating the effects of IFN-gamma. Selective gene disruption studies in mice have illustrated critical components of the host response to L. major. Mice deficient in beta 2 microglobulin, which have no major histocompatibility complex (MHC) class I or CD8+ T cells, control infection as well as wild-type mice, whereas mice deficient in MHC class II (and CD4+ T cells) suffer fatal infection. Mice with disruption of the gene coding IFN-gamma are also incapable of containing infection, reflecting absolute requirements for this cytokine. A number of interventions have been demonstrated to abrogate Th2 cell development in BALB mice, enabling these mice to control infection. Each of these--IL-12, anti-IL-4, anti-IL-2, anti-CD4 and CTLA4-Ig--has in common the capacity to make IL-4 rate limiting at the time of CD4+ cell priming. PMID- 8724834 TI - T cells and cytokines in intracellular bacterial infections: experiences with Mycobacterium bovis BCG. AB - Intracellular bacteria reside in mononuclear phagocytes, and protective immunity is dominated by T lymphocytes. Mycobacterium bovis bacillus Calmette-Guein (BCG) infection of mice represents an excellent model for studying immune mechanisms involved in defence against persistent intracellular bacteria that cause chronic disease. Gene disruption mutant mice include: A beta-/-, which lack conventional CD4+ T cell receptor alpha/beta (TCR alpha/beta) T lymphocytes; beta 2 microglobulin -/-, which lack conventional CD8+ TCR alpha/beta lymphocytes; TCR beta-/-, which lack all TCR alpha/beta lymphocytes; TCR delta-/-, which lack all TCR gamma/delta lymphocytes; and RAG-1-/- mutants, which lack mature T and B lymphocytes. Studies of these mutants suggest that CD4+ TCR alpha/beta, CD8+ TCR alpha/beta and TCR gamma/delta T lymphocytes all contribute to immunity against M. bovis BCG. Activation of antibacterial effector functions in macrophages by T helper 1 (Th1) cell-derived gamma-interferon (IFN-gamma) is central to protection. In contrast, Th2 cells are only marginally involved. Activation of Th1 and Th2 cells is regulated by interleukin 10 (IL-10) and IL-12, which are induced early in infection with M. bovis BCG. Although IL-12 is stimulated by M. bovis BCG in immunocompetent mice, studies with IFN-gamma receptor-deficient and tumour necrosis factor alpha (TNF-alpha) receptor-deficient mutant mice suggest that M. bovis BCG-induced IL-12 secretion depends on IFN-gamma and TNF-alpha. Hence, IL-12 cannot be the first cytokine produced during M. bovis BCG infection. PMID- 8724835 TI - Cytokines in immune regulation/pathogenesis in HIV infection. AB - Two hallmarks of immunopathogenesis in the progression of HIV-infected individuals to AIDS are the loss of T helper (Th) cell function in response to antigens and the critical reduction in CD4+ T cell numbers. It is probable that these two phenomena are related. We observed that: (1) the failure to detect antigen-stimulated Th cell responses in vitro correlates with increased pokeweed mitogen/staphylococcal enterotoxin B (P/S)-stimulated and antigen-stimulated T cell death; and (2) both of these events are similarly modulated by immunoregulatory cytokines. Interleukin 2 (IL-2) and IL-12 (Th1-type cytokines), as well as antibodies to IL-4 and IL-10 (which are Th2-type cytokines) restore in vitro Th cell responses to recall antigens such as influenza virus and HIV envelope synthetic peptides (env). P/S-induced T cell death affects both CD4+ and CD8+ T cell subsets, whereas death induced by stimulation with env affects only CD4+ T cells. In both examples, Th1-type cytokines and antibodies to Th2-type cytokines protect against T cell death. In contrast, IL-4 and IL-10 do not protect against death, and anti-IL-12 antibody can enhance T cell death. Our findings indicate that the loss of Th cell function and increased T cell death seen in vitro are correlated, and that in vivo HIV infection gives rise to inappropriate cytokines resulting in immune dysfunction and immunopathogenesis. PMID- 8724836 TI - Regulation of CD4+ T cell differentiation. AB - Naive T cells can be induced to differentiate from an uncommitted precursor to T helper 1 (Th1) and Th2 cells. During this differentiation, genes for transcription factors are activated, and transcription factors such as AP-1 accumulate. To study this activation, we have developed reporter transgenic mice for a number of factors, including AP-1. Naive T cells require two signals to activate AP-1. However, upon becoming effector cells, activation through diacylglycerol analogues is sufficient. Rested effector cells lose accumulated AP 1, and the induction of AP-1 synthesis requires both Ca2+ diacylglycerol signals, but not co-stimulation. PMID- 8724837 TI - The role of subsets of CD4+ T cells in autoimmunity. AB - It is generally considered that T cells which are reactive with self-antigens are effectively eliminated by two processes: clonal deletion and the induction of T cell anergy. More recently, it has been shown that some potentially autoreactive T cells remain unactivated because the self-antigens for which they are specific are not presented on competent antigen-presenting cells. All these mechanisms of self-tolerance may be regarded as passive in the sense that the autoreactive cells are either deleted or are intrinsically non-responsive. If this view of self-tolerance is adopted, then one would predict that rendering animals relatively lymphopoenic should not give rise to autoimmune disease. This prediction is not verified by experiment. Rats rendered relatively lymphopoenic by adult thymectomy followed by repeated low dose gamma-irradiation develop a high incidence of autoimmune diabetes. Furthermore, it has been shown that the reconstitution of these rats with a specific subset of CD4+ T cells from syngeneic donors prevents the development of this disease. The protective cells have the CD45RClow phenotype, they are resistant to adult thymectomy and the majority of them appear to be non-activated in the donor rats. In contrast, the CD45RChigh CD4+ subset does not provide protection from diabetes. Instead, on injection into athymic rats, it gives rise to pathological changes in a variety of organs: stomach, pancreas, liver, thyroid and lung. In addition, the CD45RClow CD4+ subset prevents these manifestations of autoimmunity in these circumstances. Recently, we have shown that CD4+ CD8- thymocytes are a highly potent source of cells that have the ability to control autoimmune diabetes in rats. It appears that the thymus has three distinct functions: positive selection; negative selection; and the generation of a population of cells that seem specialized for the control of autoimmunity. PMID- 8724838 TI - Signalling events in the anergy induction of T helper 1 cells. AB - T cells can interact productively with altered peptide ligands (APLs) resulting in different phenotypic outcomes. Stimulation of T helper 1 cells with an APL on live antigen-presenting cells results in the induction of anergy. We investigated the intracellular signalling events involved in generating this anergy by comparing protein tyrosine phosphorylation patterns after stimulation with the anergy-inducing APL or the immunogenic peptide. Stimulation by an APL resulted in a unique pattern of T cell receptor (TCR) phospho-zeta species, which was not observed with any dose of immunogenic peptide. This altered phospho-zeta pattern had a profound functional significance, in that the tyrosine kinase ZAP-70 was not activated. Thus, anergy can be induced by changing the constellation of intracellular signalling events in a T cell. These findings demonstrate that the TCR-CD3 complex can engage selective intracellular biochemical signalling pathways as a direct consequence of the nature of the ligand recognized and the initial phosphotyrosine pattern of the TCR-CD3 proteins. This then leads to different phenotypes. PMID- 8724839 TI - The two faces of interleukin 12: a pro-inflammatory cytokine and a key immunoregulatory molecule produced by antigen-presenting cells. AB - Interleukin 12 (IL-12) is produced by phagocytic cells, antigen-presenting cells and B lymphocytes in response to bacteria or intracellular parasites. IL-12 acts on T and natural killer (NK) cells inducing: production of cytokines, particularly gamma-interferon (IFN-gamma); proliferation; and enhancement of cell mediated cytotoxicity. Early in infection, IL-12 acts as a proinflammatory cytokine and induces IFN-gamma production by NK and T cells. IFN-gamma activates the phagocytes and increases their ability to produce IL-12. Unlike IFN-gamma, IL 10, IL-4, IL-13 and transforming growth factor beta are negative regulators of the production and activity of IL-12. IL-12 sets the stage for the ensuing adaptive immune response by stimulating the generation of T helper 1 (Th1) cells. It is likely that the balance between IL-12 (favouring a Th1 response) and IL-4 (favouring a Th2 response) determines the eventual outcome of the Th1/Th2 dichotomy during an immune response. HIV-infected patients have a deficient production of IL-12, even at early stages of the disease. However, exogenous IL 12 can improve the deficient immune responsiveness of these patients' T and NK cells in vitro, suggesting a possible role of the IL-12 deficiency in HIV disease pathogenesis and a potential therapeutic role of IL-12 both against opportunistic pathogens and HIV infection itself. PMID- 8724840 TI - Interleukin 15 and its receptor. AB - Interleukin 15 (IL-15) is a member of the four-helix bundle cytokine family that shares many in vitro biological activities with IL-2. Previous work demonstrated that IL-15 utilizes the beta and gamma chains of the IL-2 receptor (IL-2R), and that these are essential for IL-15-mediated signal transduction. However, several lines of evidence indicated the existence of an additional, IL-15-specific receptor component. An IL-15 binding chain was identified on a murine T cell clone, and direct expression cloning was used to isolate the corresponding cDNA. The predicted structure of this protein shows sequence similarity to the IL-2R alpha chain. Transfection of this cDNA into a murine, IL-3-dependent myeloid cell line, 32D-01, conferred IL-15 binding and, together with transfection of the IL 2R beta chain, rendered the cells responsive to IL-15 stimulation. This experiment confirmed that the IL-15 binding chain is part of the IL-15 receptor, and it is designated as the IL-15R alpha subunit. The expression pattern of the IL-15R alpha mRNA is distinct from that of IL-2R alpha mRNA. Recombinant expression of a soluble form of IL-15R alpha demonstrated that it is a potent inhibitor of IL-15 biological activity. PMID- 8724841 TI - Nitric oxide in infectious and autoimmune diseases. AB - Nitric oxide (NO) is a critical mediator of a variety of biological functions. A range of micro-organisms, including viruses, bacteria, protozoa and helminths, is sensitive to NO produced by macrophages activated with gamma-interferon (IFN gamma) and lipopolysaccharide. In contrast, NO is involved in a number of important immunopathologies, including diabetes, graft-vs-host reaction, rheumatoid arthritis, systemic lupus erythematosus, experimental autoimmune encephalomyelitis and multiple sclerosis. Thus, it is crucial that the synthesis of NO is under tight regulation. This is achieved, in part, through the opposing cytokines produced by T helper 1 (Th1) and Th2 cells. Th1 cells produce IFN gamma, which is the most powerful inducer of inducible NO synthase (iNOS). In contrast, interleukin 4 is produced by Th2 cells and inhibits the induction of iNOS at the level of transcription. Furthermore, NO is also produced by Th1 cells, whose proliferation can be inhibited by high concentrations of NO. Thus, apart from being a mediator of Th1/Th2 interaction, NO may also be an important self-regulatory molecule that prevents the over-expansion of Th1 cells which are implicated in a range of severe immunopathologies. PMID- 8724842 TI - Ambulatory elderly patients of primary care physicians: functional, psychosocial and environmental predictors of need for social work care management. AB - With increasing numbers of elderly people, and the escalating costs of health care, screening becomes increasingly important for identifying those older people with social health care needs who appear in their primary care physicians' offices. Many people are not aware of available social services. Families with serious social problems are not finding the help they need. The aim of this study was to develop and refine a questionnaire as a screening tool to identify elderly outpatients in primary care settings who are at high risk for psychological, social or environmental needs. This study identified those ten factors at each site which were most indicative that further intervention was needed. There were consistencies among the coordinators across sites in terms of what factors triggered intervention. Activities of Daily Living (ADLs) and Instrumental Activities of Daily Living (IADLs) were more likely to be predictive of the coordinator's intervention than were other factors. PMID- 8724844 TI - Clients' views of social work services in the hospital setting in Israel. AB - Client appraisals of social work services are likely affected by the setting in which care is offered. This paper examines clients' assessments of social work services received in the acute care hospital setting. It is based on interviews with 120 discharged hospital social work clients, and investigates their definition of their psychosocial problems, their expectations from the social work department and their appraisal of the outcomes of the social work intervention. Main findings include the dominance of instrumental needs among the psychosocial problems cited by clients, while emotional relief was the most prevalent outcome. A general lack of clarity regarding expectations was observed. The findings are discussed in light of organizational and methodological issues which might influence the examination of clients' assessments. PMID- 8724843 TI - Research changes a health care delivery system: a biopsychosocial approach to predicting resource utilization in hospital care of the frail elderly. AB - The social work department of a large New York City teaching hospital has conducted practice research studies over the past five years to better improve the early identification of high risk elderly patients. A recent prospective study is presented of the relationship between functional capacity and discharge status/length of stay, the results of which directly led to a change in the inpatient delivery system. A sample of 250 randomly selected patients, 65 years or older, selected upon admission, were tested using the Katz ADL, the IADL and the Short Portable Mental Status Questionnaire (PMSQ) to distinguish those patients likely to remain hospitalized beyond medical necessity. Results of a logistic regression indicate that low functional capacity patients (IADL: Wald = 5.6; P < .02) were likely to remain beyond medical necessity. The model predicted group membership correctly 90.6% of the time. These findings have led the medical center to develop an acute care geriatric unit in cooperation with an affiliated nursing home. PMID- 8724845 TI - Interdisciplinary teams in health care: integration of family caregivers. AB - Interdisciplinary health care teams function within a range of contexts, have varying structures, and engage in different processes, which result in a range of outcomes for patients as well as for the team as a whole. Each of the components highlighted is influenced by the extent to which patients and their family caregivers are involved in the functioning of teams providing care. A model detailing each highlighted component of team functioning is presented. The integral roles of patients and their caregivers within teams are emphasized in terms of the impact on team functioning. PMID- 8724846 TI - The advocacy role in hospital social work. AB - Hospital social workers were asked to describe their roles and functions, with particular emphasis on the amount of time spent doing traditional advocacy on behalf of patients. The results indicated that workers spent the least amount of time on advocacy as compared with other traditional social work roles. A case is made for emphasizing the advocacy role as a strategy to counteract the potentially negative effects on social work of organizational changes in Canadian hospitals. The implications of the findings for schools of social work, professional associations and employers of social workers are discussed. PMID- 8724847 TI - An expression profile of active genes in cultured human keratinocytes. AB - An expression profile of genes active in cultured human keratinocytes was obtained by collecting 770 partial sequences from a 3'-directed cDNA library that faithfully represents the mRNA population in the source cells. Ninety-four species composed of 258 clones occurred recurrently, and 512 clones appeared only once. The gene which showed the most abundant expression codes for type I keratin 14, the major keratin that is known to be strongly expressed in the basal layer of the epidermis. Altogether 242 gene species were identified in GenBank, 9.5% of which encode cytoskeletal proteins and 14.5% the components of protein synthesis. Keratin synthesis, the unique property of the keratinocyte, has been measured by elevating the level of transcription of several keratin genes. However, there was no expression of the differentiation markers, type II keratin 1 (K1) or type I keratin 10 (K10), indicating that the cells used for our cDNA library construction were in the pre-differentiation stage. By comparing this expression profile with seven expression profiles from other tissues/cells, four clones, including a novel clone were tentatively identified as specific to keratinocytes. PMID- 8724848 TI - Identification of a novel human gene containing the tetratricopeptide repeat domain from the Down syndrome region of chromosome 21. AB - The Down syndrome (DS) region on chromosome 21, which is responsible for the DS main features, has been defined by analysis of DS patients with partial trisomy 21. Within the DS region, we constructed a 1.6-Mb P1 contig map previously. To isolate gene fragments from the 1.6-Mb region, we performed direct cDNA library screening and exon trapping using the P1 clones and a human fetal brain cDNA library, and obtained 67 cDNA fragments and 52 possible exons. Among them, 23 cDNA fragments and 4 exons were interpreted to be derived from a single gene by localization on P1 clones and by Northern analysis. To obtain the full-length cDNA sequence, longer cDNA clones were further screened from another human cDNA library which was enriched with longer cDNA species. These clones were sequenced and assembled to a sequence of 9045 bp. This transcribed sequence encodes a novel 2025 amino-acid protein containing tetratricopeptide repeat (TPR) motifs and therefore the gene was designated as TPRD (a gene containing the TPR motifs on the Down syndrome region). The TPR domain has been found in a certain protein phosphatase and in other proteins involved in the regulation of RNA synthesis or mitosis. The TPRD gene, the novel gene which was proved to be in the 1.6-Mb region and to have the interesting features described above, is a candidate for genes responsible for the DS phenotypes. PMID- 8724849 TI - Prediction of the coding sequences of unidentified human genes. V. The coding sequences of 40 new genes (KIAA0161-KIAA0200) deduced by analysis of cDNA clones from human cell line KG-1. AB - As part of our continuing efforts to accumulate information on the coding region of unidentified human genes, we newly determined the sequences of 40 cDNA clones of human cell line KG-1 which correspond to relatively long and nearly full length transcripts, and predicted the coding sequences of the corresponding genes, named KIAA0161 to 0200. The average size of the cDNA clones analyzed was approximately 5.0 kb. A computer search of the sequences in public databases indicated that the sequences of 20 genes were unrelated to any reported genes, while the remaining 20 genes carried sequences which show some similarities to known genes. Among the genes in the latter category, KIAA0167 contained a Zn finger motif with significant structural similarity to that of the yeast transcription factor GCS1, and KIAA0189 was classified into the RhoGAP gene family. Stretches of typical CAG (Gln) repeats, which were often correlated with genetic disorders, were found in KIAA0181 and KIAA0192. Another novel repeat composed of alternating Arg and Glu was identified in KIAA0182. Northern hybridization analysis demonstrated that 10 genes are expressed in a cell- or tissue-specific manner. PMID- 8724850 TI - Conservation and periodicity of DNA bend sites in eukaryotic genomes. AB - DNA bend sites appear every 680 bp on average in the human epsilon- and beta globin gene regions. Although most of their molecular nature has not been unraveled, a potential bend core sequence A2N8A2N8A2 (A/A/A) and its complementary T2N8T2N8T2 (T/T/T) appeared preferentially either in or very close to most of the bend sites, whereas other combinations of A2 and T2 dinucleotides, A/T/T + A/A/T, T/T/A + T/A/A and A/T/A + T/A/T, did not. The distances between any two of the core sequences in the entire beta-globin locus showed a strong bias to a length of 701-800 bp and multiples thereof, suggesting that there is periodicity throughout the locus. This bias was not found for other combinations of A2 and T2. Again, this periodicity was identified in many eukaryotic genes, whereas the tendency was absent in mRNAs and prokaryotic as well as viral genomes. PMID- 8724851 TI - The nucleotide sequence of human acylamino acid-releasing enzyme. AB - The nucleotide sequence of a cDNA coding for the human acylamino acid-releasing enzyme (AARE, also known as acylpeptide hydrolase) [EC 3.4.19.1] subunit has been determined. The amino acid sequence of human AARE subunit deduced from its cDNA nucleotide sequence showed a high degree of identity (91.5%) with both the corresponding proteins from the pig and the rat. The AARE cDNA shows 99.2% identity with a 3.3 kb cDNA transcribed from a locus (DNF15S2) on the short arm of human chromosome 3, whose deletion is associated with small cell lung cancer, taking into consideration that the sequence of the 3.3-kb cDNA previously reported was caused by misreading. PMID- 8724852 TI - An improved cosmid vector for the nested deletion method using the bacteriophage T3 DNA packaging system. AB - We constructed a new cosmid vector suitable for the previously developed nested deletion method which used the in vitro DNA packaging system of bacteriophage T3. The first step of this method is linearization of a cosmid clone to be packaged, and we previously introduced cleavage at the cos site using lambda-Terminase, but optimization of the reaction conditions was required for complete digestion because of its instability. In the newly constructed vector, pAT5, the sites of 4 different restriction enzymes, Sse8387I, Asc I, Fse I and Pme I, each of which recognizes an 8-bp sequence (8-base cutter) were introduced in the vicinity of the cos site. In addition, the species of restriction sites for cloning were increased to broaden its application. The cosmid clone constructed by this new vector could be linearized at one of the 8-base cutter sites which are assumed to rarely occur in the genome, and followed by in vitro packaging, nested deletion clones were successfully prepared. PMID- 8724853 TI - Prediction of the coding sequences of unidentified human genes. V. The coding sequences of 40 new genes (KIAA0161-KIAA0200) deduced by analysis of cDNA clones from human cell line KG-1 (supplement). PMID- 8724854 TI - Effect of interferon-alpha on the steady-state levels of thymine, thymidine, guanine, and guanosine metabolites in herpes simplex virus-type 1-infected cells. AB - Interferon-alpha (IFN-alpha) combined with acyclic guanine analogs synergistically inhibits replication of herpes simplex virus type 1. IFN-alpha treatment influenced the metabolism of exogenously supplied nucleobases and nucleosides in a manner expected to contribute to synergistic activity. IFN-alpha treatment of infected human cornea stromal cells or Vero cells significantly reduced steady-state levels of acid soluble metabolites of thymine, as well as thymidine, that accumulate early during virus replication but did not affect metabolism of thymine and thymidine in uninfected cells. IFN-alpha treatment significantly reduced the ability of uninfected cells to accumulate acid-soluble metabolites from guanine, but not guanosine. The effects of IFN-alpha on nucleobase/nucleoside metabolism could contribute to synergistic antiviral activity by reducing the accumulation of thymidine/thymine metabolites and decreasing the guanine taken into cells. PMID- 8724855 TI - Characteristics of single- and double-stranded RNA synthesis by a rotavirus SA-11 mutant thermosensitive in the RNA polymerase gene. AB - The phenotype of a rotavirus SA-11 mutant, ts C, which carries a mutation in the gene coding for the viral RNA polymerase was studied in vitro. ts C viral transcription proved to be sensitive to temperature in a different way to that previously described. Like the wild type, the ts C mutant has an optimum for in vitro transcription at 45 degrees, but mRNA synthesis was inhibited at temperatures over 50 degrees. This mutant also showed a higher resistance to transcriptional inhibition by nucleotide analogues than the wild-type strain. The in vitro minus-strand RNA synthesis catalysed by ts C particles indicates that the mutant does not exhibit the expected increased sensitivity to temperatures over 31 degrees shown by the in vivo phenotype. As with plus-strand synthesis, the optimal temperature for the minus-strand synthesis assay was 45 degrees, but for temperatures over 55 degrees, the number of double-stranded RNA products was altered. Our results suggest that when in vitro plus- and minus-strand RNA synthesis in ts C and wild type are compared, the mutated VP1 motif affects both transcription and minus-strand synthesis, but in different ways. In infected cell cultures, the results also show that the phenotype associated with ts C seems to mainly affect the function of plus-strand RNA synthesis. PMID- 8724856 TI - Primary keratinocytes can be infected by natural isolates of genital human papillomavirus. AB - Viral particles of human papillomavirus (HPV) types 11 and 16 were isolated from flat condylomas and biopsies of dysplastic lesions and used to infect human keratinocytes. The presence of HPV DNA in infected cells was determined by molecular hybridization and the polymerase chain reaction. Late transcripts were detected shortly after infection. The persistence of HPV was limited to early passages after infection and the loss of viral DNA preceded cell culture senescence. Our experimental evidence supports the idea that pooling of several lesions can lead to the isolation of infectious HPV 16 virions. PMID- 8724857 TI - Antibody responses to Epstein-Barr virus, human herpesvirus 6 and human herpesvirus 7 in patients with chronic fatigue syndrome. AB - To test for an association between chronic fatigue syndrome (CFS) and infections with Epstein-Barr virus (EBV), human herpesvirus 6 (HHV-6) and human herpesvirus 7 (HHV-7), antibodies to these viruses were tested in the serum from three groups of individuals: (1) 10 CFS patients with chronic fatigue beginning with a clinical pattern of acute infectious mononucleosis [IM; true chronic IM (CIM)]; (2) 10 CFS patients whose illness did not start with acute IM (non-CIM), and (3) healthy controls. High EBV antibody titers were demonstrated in most patients. Antibodies to ZEBRA, a product of the immediate early EBV gene BZLF1, were detected in the serum of CFS patients at a higher frequency than in healthy controls. Antibody titers to HHV-6 and HHV-7 were also higher in the patients with CFS than in the controls. These results are consistent with the view that CFS patients may have reactivations of EBV, HHV-6 and HHV-7. PMID- 8724858 TI - The relationship of Piry virus to other vesiculoviruses: a re-evaluation based on the glycoprotein gene sequence. AB - By sequencing the 3 half of the Piry virus genome, we show that Piry virus, like the other vesiculoviruses, contains the genes for nucleoprotein N, phosphoprotein P, matrix protein M, glycoprotein G and polymerase protein L, in that order. Our analysis of the Piry G protein sequence suggests that Piry and Chandipura are related to each other as closely as the Indiana and New Jersey vesicular stomatitis virus serotypes are to each other. A re-examination of amino acid sequences in the nucleocapsid protein shows that this relationship is also true of the more conserved central region of this protein and that the greatest divergence between Piry and Chandipura has occurred in two other regions of the nucleocapsid protein. PMID- 8724859 TI - Amino acid substitutions in glycoprotein D mediate the ability of fusion from without-positive herpes simplex virus type 1 strains to penetrate at 4 degrees and in the presence of soluble glycoprotein D. AB - In former studies, we described herpes simplex virus type 1 (HSV-1) strains ANG, ANG path and HSZP as strains with special fusion activities, caused by mutations in the syn 3 locus. In this report, we describe the special penetration properties of these strains: their ability to penetrate at 4 degrees and to overcome the infection block caused by soluble glycoprotein D-1 (gD-1) in the medium. Using intertypic recombinants of HSV-1 strains ANG path and KOS, we showed that these penetration properties are the result of two mutations in amino acids 25 (Leu-Pro) and 27 (Gln-Arg) in the N-terminal part of the mature gD-1 glycoprotein. PMID- 8724860 TI - Mutations in the core promoter/enhancer II regions of naturally occurring hepatitis B virus variants and analysis of the effects on transcription activities. AB - The regulatory regions for transcription and replication of several hepatitis B virus (HBV) genomes from 19 patients having various forms of HBV infection were sequenced. Predominant mutations were found to occur naturally in nucleotide positions 1762 (A to T) and 1764 (G to A) in chronic hepatitis patients and in asymptomatic carriers after seroconversion, but were not observed in HBeAg positive healthy carriers. Since these positions were located in the basic core promoter and the overlapping enhancer II regions situated within the core upstream region, transcriptional activity was examined by chloramphenicol acetyltransferase assay to determine if there was a possible difference between the mutant and wild-type HBV. However, no significant difference was detected upon comparison of the promoter and enhancer activities between mutant and wild type HBV. PMID- 8724861 TI - Ribonucleoprotein complex formation by the human hepatitis B virus polymerase. AB - Human hepatitis B virus (HBV) polymerase (pol or RT), when expressed in Xenopus oocytes upon injection of synthetic minimal pol RNA (RT RNA), assembles into a higher molecular weight complex with the characteristics of a ribonuclear protein (RNP) complex. In vitro RNA competition binding data suggest that RT RNA is preferentially packaged into this complex even though it lacks the authentic viral encapsidation signal, epsilon, and viral capsid protein sequences. Consistent with this finding, the in vitro polymerase reaction performed in pol expressing oocyte extracts generates primarily HBV-specific DNAs even when the pol template is challenged with a coinjected non-HBV competitor RNA. These results suggest that interaction between pol and its cognate RNA can be mediated by sequences other than the known packaging elements. We speculate that HBV RNP complexes containing at least polymerase and viral RNA may play a role in viral nucleocapsid assembly and may help to segregate HBV reverse transcription from the cellular milieu in vivo. PMID- 8724862 TI - Chimeric BK virus DNA episomes in a papillary urothelial bladder carcinoma. AB - BK virus (BKV) DNA sequences were identified in a papillary urothelial bladder carcinoma by Southern blot hybridization. The carcinoma contained both integrated and extrachromosomal DNA. Integrated sequences had a clonal restriction pattern, suggesting that BKV was integrated at some early stage of neoplastic initiation or progression. Viral episomes consisted of a population of covalent polymers based on a high-molecular-weight DNA unit, about 11-12 kb in size. DNA sequences non-homologous to the BKV genome were encompassed within DNA episomes, suggestive of acquisition of cellular sequences by viral DNA replication at the integration site. Extrachromosomal, chimeric DNA molecules were present at an average level of about 50 copies per cell, but their size, apparently incompatible with viral assembly, showed that BKV productive infection was impaired. The data suggest that infected cells underwent reversible changes affecting autonomous BKV DNA replication. PMID- 8724873 TI - [Effect of removal of the adrenal medulla on the hormonal response induced by acute hepatic vagotomy]. AB - The purpose of the present study was to determine the effects of an acute hepatic vagotomy on plasma insulin and catecholamines levels in fasted and fed rats with and without adrenodemedullation. Rats were first divided into two groups: adrenodemedullated (ADM) and non-adrenodemedullated rats, each group being subdivided into a normally fed and a fasted (14 hrs) group. Anesthetized rats were first ADM or sham operated, and a jugular catheter was inserted. The first blood sample was taken 30 min later (time 0). Rats were then hepatic vagotomized (HV) or sham operated, and the second blood sample was taken 30 min after the HV. Results (0 vs 30 min) indicate an increase in plasma glucose, epinephrine, and norepinephrine concentrations in rats non-adrenodemedullated. This effect was independent of the nutritional state and the integrity of the vagus nerve. A weakly significant (P < 0.07) effect of the HV on insulin response was found in ADM rats, independently of the fed or fast state (different levels of hepatic glycogen content). These results indicate the necessity of using ADM animals to study the effects of an acute HV. They also suggest that the hepato-pancreatic axis is not influenced by the level of hepatic glycogen. PMID- 8724874 TI - Methodological factors affecting esophageal clearance. AB - To define some methodological factors that could affect the measurement of the esophageal clearance, we used three different protocols, on 41 healthy control subjects. In 20 subjects, we studied the influence of a naso-esophageal probe on the frequency of swallowing. We observed a significant (44 +/- 5%) decrease in the frequency of spontaneous swallows after 15 minutes, with stabilization after this 15 minute period of adaptation. In 11 subjects, we studied the influence of the pH of the esophageal content on esophageal clearance. We found an exponential relationship between the esophageal clearance (C) expressed in minutes and the initial pH of the lumen contents: C = 43.3 exp (-0.54 pH) In 10 other subjects, we studied the influence of the volume of the esophageal contents on esophageal clearance. We found no influence for volumes less than 30 ml and a significant increase of clearance for volumes greater than 30 ml. In summary, this study of the effects of methodological factors on esophageal clearance: Gives new information about the "accommodation phase" of the esophagus after distension by a naso-gastric probe; Allows us to propose a new quantitative method for evaluation of esophageal clearance Shows the small importance of the ingested volume. PMID- 8724875 TI - Effects of acute cardioselective and non-selective beta-adrenergic blockade on plasma ammonia levels in exercising dogs. AB - The aim of this study was to assess plasma ammonia levels during acute treadmill exercise in dogs after intravenous administration of a single dose of different cardioselective (atenolol) and non-selective (sotalol and propranolol) beta adrenergic blocking drugs. The experiments were performed on 6 male mongrel dogs (20-25 kg) trained to run on a motor driven treadmill. After administration of saline or atenolol, there was no significant increase in plasma ammonia during exercise; propranolol and sotalol however, both non-selective beta-blockers, produced a significant increase in plasma ammonia. Plasma levels of alanine and glutamine were not altered during exercise compared with resting values, and were not modified by the administration of beta-blockers. It is not clear whether the different effects on exercise induced hyperammonemia are due to different effects on muscle ammonia formation or on the ammonia clearance by the liver. PMID- 8724876 TI - Long-term effects on lipid metabolism in miniature swine (Sus scrofa) of diets enriched in saturated, monounsaturated and polyunsaturated (N-6 and N-3) fatty acids. AB - In this paper we study the effect of long-term adaptation (twelve months) of lipidic parameters of miniature swine to diets enriched in saturated (lard; L group), monounsaturated (olive-oil; O-group) and polyunsaturated (sunflower and fish-oil; S- and F-groups respectively). The experimental group with the highest level of total cholesterol, free cholesterol and phospholipids was the S-group. This group had as well levels of HDL-C and LDL-C significantly higher when compared with the remaining groups. The L-group had the lowest value of HDL-C. In spite of that, the index of artherogenicity (HDL/LDL+VLDL) was significantly higher in the L-group, followed by the O-, F- and S-group respectively. On the other hand, after 12 months adaptation we observed that the fatty acid composition of serum lipids clearly reflects the quality of the dietary fats. The O-group had significantly higher serum oleic acid levels than all the other groups, and its content in saturated fatty acids was the lowest. The same happens with red blood cell (Rbcs) membranes fatty acids but the effect is less marked. Membranes of the L-group were found to have the highest saturation index (SI) in Rbcs membranes, while the F-group had the highest unsaturation index (UI), followed by the O-group. Taken together, our findings show that the diet enriched in olive oil produces a lipid pattern intermediate between that obtained with the fish-oil-diet and the sunflower-oil-diet but with the advantage of Rbcs membranes with a lower amount of PUFAS. As it is known, membranes with high percentages of PUFAS are more accessible for peroxidation, and the degree of peroxidation of lipids is directly related with changes in the membranes functionality. PMID- 8724877 TI - Vascular smooth muscle reactivity and endothelium derived relaxing factor in experimental obstructive jaundice. AB - Obstructive jaundice is associated with a predisposition to systemic hypotension and acute renal failure. Altered vascular reactivity may contribute to the development of hypotension. In this experimental study on dogs, alterations in vascular contractile responses to noradrenaline, serotonin and KCl were investigated. Contractile responses to noradrenaline, serotonin,,, KCL and relaxation responses to papaverin and acetylcholine were provoked in isolated femoral arteries of both control dogs and animals with obstructive jaundice. In this situation concentration-response curves of noradrenaline and serotonin were blunted when compared with controls. This blunting disappeared when endothelium was removed. In rings precontracted with phenylephrine, EDRF relaxation responses to acetylcholine were increased significantly as compared to controls: at lower concentrations maximal relaxation response occurred. Contractile responses to KCl and relaxation responses to papaverin did not differ between the groups, endothelium present or removed. These results indicate that obstructive jaundice induces a decrease in vascular contractile responses and an increased EDRF relaxation response. We suggest that an excess in the amount of released EDRF may be one of the causes inducing systemic hypotension in obstructive jaundice. PMID- 8724878 TI - Lipid and fatty acid contents in red tides from tropical fish ponds of the coastal water of South China Sea. AB - Microplanktonic red tide blooms (dominated by dinoflagellates) were observed in brackish water fish ponds of Terengganu between March 1992 to January 1993. The first short-lived bloom (2-3 days) occurred in October 1992 while the second long lived bloom (6-7 days) occurred in January 1993. The dominant dinoflagellate species comprised of Peridinium quinquecorne (> 90% total cell count) with considerable proportion of Protoperidinium excentricum. Ciliophora consisting of Tintinopsis sp. and Favella sp. were also present during the bloom period. The total ash, chlorophyll, phaeopigment, lipid and fatty acid content of the microplankton were studied. Considerable amounts (6-11% of the total fatty acid) of the polyunsaturated fatty acid 18:3w3 (linolenic acid) were present in the microplankton. However, high amounts of 20:5w3 (eicosapentanoic acid) and 22:6w3 (docosahexaenoic acid) were present with variable but usually high amounts of 22:4w6 and 22:5w6 acids. The latter microplankton bloom contained higher amounts of 20:5w3 and 22:6w3 acids than the earlier bloom. Lipid content were three to five times higher than chlorophyll a. There was an increase with successive day after bloom outbreak in the relative proportion of total C18, C20, and C22 fatty acid components. The algae microplankton contained the w3-polyunsaturated fatty acids (PUFAs) probably needed for the growth and survival rate of grazing pond animals. PMID- 8724880 TI - Electrical activity recorded from abdominal surface before and after gastric surgery in man. AB - To explain the role of the different portions of the stomach in the genesis of electrical signal obtained from abdominal surface (Electrogastrography, EGG), the EGG was recorded in 6 patients with gastric cancer and 7 with duodenal ulcer before and after total gastrectomy or Billroth II gastric resection, respectively. Eight patients undergoing cholecystectomy entered the study as controls. The spectral frequency components were subdivided into ranges, and the dominant frequency and power were calculated for each range before and after surgery. The power profile, expressed as difference in power percentages before and after surgery for each frequency range, was obtained. The power profile from total gastrectomy or gastric resection was clearly different from that resulting from cholecystectomy in the 2.6-3.5 cycles per minute range (cholecystectomy vs gastric resection, P = 0.009; cholecystectomy vs gastrectomy, P = 0.012). No difference in power profile between total gastrectomy and gastric resection was demonstrated. Since total and partial gastrectomized patients showed a similar power profile, EGG signal corresponded to the electrical activity of the distal two third of the stomach. PMID- 8724879 TI - [Effect of the nature of the heteroatom of a monosaccharide derivative on its inhibitory action in relation to P-type calcium channels expressed in the Xenopus oocyte]. AB - Our results demonstrate that saccharidic derivatives obtained by adding a C8 alkyl group through various heteroatomes (O, N or S) to a monoacetonide residue possess an inhibitory effect towards putative P-type calcium channels expressed in Xenopus oocytes. These derivatives partially and reversibly inhibit the activity these channels without changing their electrophysiological properties. Nevertheless, the derivative containing the heteroatome N also affects the fast and tetrodotoxin-sensitive sodium channel activity. Thus, only ether and thioether compounds (heteroatome O or S) can be selected for their inhibitory effect on P-type apparented calcium channels. PMID- 8724881 TI - Ca2+ entry through acetylcholine receptor channel in dysgenic myotubes. AB - Skeletal muscles of mutant mice with "muscular dysgenesis" are characterized by excitation-contraction uncoupling resulting from the absence of dihydropyridine receptors. However contraction of the dysgenic myotubes can be evoked by afferent nerve stimulation or by ionophoretic application of acetylcholine (ACh) on the muscle. These contractions are elicited by Ca2+ entry through the ionic channel of the ACh receptor at multiple synaptic contacts. In the present paper, the calcium entry through ACh receptors was compared in cultured normal and dysgenic myotubes. At elevated external calcium concentration (110 mM), the elementary slope conductance of the ACh-activated ionic channel of dysgenic myotubes did not differ from that found in normal myotubes. We conclude that dysgenic muscle contraction induced by nerve stimulation does not result from an abnormal Ca2+ entry across ACh receptors. We discuss the possible involvement of sustained high threshold calcium current (Idys) and of the calcium induced calcium release mechanism in the contractile response related to synaptic activity of dysgenic myotubes. PMID- 8724882 TI - [Monitoring of the permeability of the inferior esophageal sphincter using sound signals]. AB - An original method is described for graphic display of sounds arising from the cardia during the swallowing of saliva or water. Two dynamic microphones were placed on the skin of epigastric xyphoid area and simultaneously on the latero cervical position. The following conditions must be filled: position of the subject (upright or lying), correct application of microphone with pressure (2 kPa), specially for the xyphoid situation. For graphic restitutions from magnetic signal, a time-recording paper was used, on line or off line, with an analogical device including Butterworth band active filter (optimal frequency range 0.8-1.2 kHz), amplifier and integrator (tau = 2 s). Occurrence of typical cardia sounds were 86-97 % for saliva and 95-100 % for water in two normal subjects for the evaluation of reproducibility (one man and one woman 35 and 47 years old respectively, n = 234 sequences, interval 20 s). Using the distance between the upper and lower microphones, the esophageal time transit can be measured. The signal was more unconstantly recorded for saliva 40 +/- 44 % in adults (n = 10) and 76 +/- 17 % in children (n = 7), than for 80 et 87 % for water respectively. The optimal interval between two swallows appeared to be 15-25 s. In patients with presumption of achalasia (n = 8), the two phonic signals were recorded during esophageal pressure measurement. The presence of esophageal sounds seems to be the sign of absence of achalasia. PMID- 8724883 TI - Relationship between heart function and energy production. A study on isolated rat heart. AB - The purpose of this study is to determine the relationship between cardiac performance and energy production in isolated rat heart when heart function is modified either by calcium concentration or by oxygen partial pressure (PO2), and to evaluate the relative contribution of glycolytic ATP. Hearts are perfused at a constant 10 ml/min flow and submitted to increasing calcium concentration (0.36 to 1.78 mM free calcium) with maximal PO2 or to graded hypoxia (660 to 52 mmHg) with maximal calcium concentration. Cardiac performance, oxygen consumption (VO2), lactate+pyruvate production are measured. To inhibit glycolysis, perfusions are also carried out with deoxyglucose (2-DG). The plotting of mitochondrial ATP production, as calculated from VO2 vs contractility parameters shows a different relationship when we modify the PO2 or the calcium concentration, whereas the relationship is similar for heart rate. When cardiac performance is related to total ATP production, glycolytic ATP being calculated from lactate+pyruvate production, the difference, although decreased, remains. 2 DG impairs heart function, but with 2-DG the relationship between ATP production and heart function becomes unique. In conclusion, there is an evident difference in the dependence of heart contractility on ATP production according to the factor that limits heart function. The contribution of glycolysis to energy production does not explain all of this difference. Furthermore, such a difference does not exist for heart rate. This raises the question of energy compartmentation in myocardial cells. PMID- 8724884 TI - Adrenocortical response to open-field test in rats with anterodorsal thalami nuclei lesion. AB - The influence of limbic anterodorsal thalami nuclei (ADTN) on adrenocortical activity and on emotional reactivity were investigated in male and female rats. The emotional reactivity was evaluated by means of the open-field test and the corticoadrenal function by means of plasma and adrenal corticosterone concentration. The results demonstrate that ADTN lesion does not affect the behavioural patterns in the open-field test on the 29th and 30th day after lesion nor adrenal response when animals are exposed to a novel situation. PMID- 8724885 TI - Hemodialysis vascular access morbidity. AB - Complications associated with hemodialysis vascular access represent one of the most important sources of morbidity among ESRD patients in the United States today. In this study, new data on the magnitude and growth of vascular access related hospitalization in the United States is presented, demonstrating that the costs of this morbidity will soon exceed $1 billion per yr. This study also reviews published literature on the morbidity associated specifically with native arteriovenous fistulae, polytetrafluoroethylene bridge grafts, and permanent central venous catheters. Next, new information on the changing patterns of vascular access type in the United States is presented, demonstrating the continuing evolution of medical practice away from the use of arteriovenous fistulae in favor of more reliance on synthetic bridge grafts. Based on these data, a discussion is provided of the tradeoffs among the most commonly available modalities of vascular access today. Although radial arteriovenous fistulae continue to represent the optimal access modality, the appropriate roles for brachial arteriovenous fistulae, synthetic bridge grafts, and central venous catheters are less certain because of inadequate data on the long-term function of the first and the high rates of complications associated with the latter two. To reduce vascular access-related morbidity, strategies must be developed not only to prevent and detect appropriately early synthetic vascular access dysfunction, but to better identify the patients in a whom radial arteriovenous fistula is a viable clinical option. PMID- 8724886 TI - Antibiotic-resistant endocarditis in a hemodialysis patient. AB - A chronic dialysis patient developed persistent bacteremia as a result of infection with Enterococcus faecium. During the patient's illness, resistance to ampicillin, gentamicin, vancomycin, and teicoplanin developed. Despite arteriovenous (AV) graft removal and an extensive but inconclusive search for the source of the infection, bacteremia persisted. On autopsy, the patient was found to have had aortic-valve endocarditis. Endocarditis is a well-known complication in dialysis patients. Multidrug-resistant organisms are becoming more prevalent in hospitalized patients as well. Risk factors for the development of endocarditis in dialysis patients include catheters, AV grafts, and calcific valvular disease, all in conjunction with frequent access to the circulation. Avoidance of temporary catheter use by prompt placement of AV fistulas or grafts and consideration of their early use, the meticulous care of catheters once in place, and treatment of the nasal carriage of Staphylococcus aureus may lower the incidence of bacteremia and therefore endocarditis in dialysis patients. The removal of infected catheters and/or AV grafts if prompt clearing of the blood with antibiotics does not occur is the next step, followed by valve replacement in selected cases. The routine use of vancomycin in the dialysis population should be reevaluated in light of the development of high-level antibiotic resistant organisms. PMID- 8724887 TI - Long-term cyclosporine therapy for pediatric nephrotic syndrome: a clinical and histologic analysis. AB - Cyclosporine (CsA) is effective in treating steroid-dependent (SDNS) and steroid resistant (SRNS) nephrotic syndrome (NS) in children, but because of the potential for chronic nephrotoxicity, its long-term use is controversial. This study reports the results of long-term CsA treatment in 22 children with idiopathic NS. Indications for treatment included SDNS (N = 7) and SRNS (N = 15) children. Pre-CsA histology showed minimal change disease in three patients, immunoglobulin M nephropathy (IgM) in 14 patients, and focal segmental glomerulosclerosis (FSGS) in five patients. All patients had normal initial serum creatinine values. CsA was added to prednisone at 6.3 +/- 0.4 mg/kg per day (mean +/- SE) and adjusted to maintain whole blood trough HPLC levels of 70 to 120 ng/mL for a period of 6 to 53 months (mean, 22 months). Analysis by clinical course revealed that 13 of 15 patients with SRNS (87%) entered remission after a mean duration of CsA treatment of 58 days, whereas seven of seven patients with SDNS were able to be weaned off of daily prednisone therapy. Histologic analysis showed that all five patients with FSGS and 13 of 14 patients with IgM nephropathy either entered remission or were weaned off of daily steroids. Ten of the 22 patients (45%) with complete remission required CsA plus low-dose alternate-day prednisone to maintain remission. Hypertension was seen in eight of 22 patients (36%). No patient had a significant increase in serum creatinine concentration. Renal biopsies performed in 12 patients after 12 to 41 months (mean, 21 months) of CsA therapy showed no nephrotoxicity or disease progression in ten patients. Progression of the previous interstitial fibrosis and tubular atrophy was noted in two patients, suggesting a 17% incidence of CsA nephrotoxicity. This analysis of the long-term risks and benefits of CsA for childhood NS has identified two important findings: (1) combined CsA and alternate-day steroids can be highly effective in inducing complete remission in patients with SRNS and biopsy-proven IgM nephropathy, and (2) long-term use of CsA in moderate doses with closely monitored levels can result in a relatively low incidence of nephrotoxicity. PMID- 8724888 TI - Higher maintenance cyclosporine dose decreases the risk of graft failure in North American children: a report of the North American Pediatric Renal Transplant Cooperative Study. AB - Because of changing cyclosporine dosing patterns over the years, maintenance doses in 1469 living donor and 1486 cadaver donor index renal transplants in patients who were 0 to 20 yr of age of the time of transplantation were reviewed. All grafts had at least 30 days' function. Eighty-nine percent of living donor patients and 95% of cadaver donor patients were maintained on cyclosporine for 12 months after transplantation. Among patients receiving cyclosporine, the maintenance 12-month dose in living donor graft recipients increased from 6.4 mg/kg per day for patients who had transplants done in 1987 to 7.9 mg/kg per day for patients who had transplants done in 1992 (P = 0.02). Among cadaver donor graft recipients, the mean 12-month maintenance dose increased from 6.4 mg/kg per day in 1987 to 7.8 mg/kg per day in 1992 (P = 0.01). At 6 months after transplantation, the maintenance cyclosporine dose in 1103 living donor graft recipients who retained graft function during the subsequent 6-month period was 7.1 mg/kg per day, compared with a 6-month dose of 3.9 mg/kg per day in 28 patients who lost their graft in the subsequent 6-month period. For 1041 recipients of cadaver donor grafts who retained graft function through 12 months after transplantation, the 6-month maintenance cyclosporine dose was 7.4 mg/kg per day, compared with 5.4 mg/kg per day in 59 patients whose graft failed in the subsequent 6 months. In patients who did not have an acute rejection episode during the first 12 months after transplantation, the rate of subsequent "late" rejections was 22% for patients with a 12-month maintenance cyclosporine dose < or = 4.0 mg/kg per day, and 16% for patients whose 12-month maintenance cyclosporine dose exceeded 8.6 mg/kg per day. A proportional hazards regression analysis, using cyclosporine dose as a time-dependent covariate, showed that the hazard of graft failure was reduced 5 to 6% for each incremental increase of 1 mg/kg maintenance dose of cyclosporine (within the dose range studied) for both living and cadaver donor source transplants. PMID- 8724889 TI - Effects of diet and antihypertensive therapy on creatinine clearance and serum creatinine concentration in the Modification of Diet in Renal Disease Study. AB - Factors other than the glomerular filtration rate (GFR) can affect creatinine clearance (Ccr) and serum creatinine concentration (Pcr). The effect of dietary protein and antihypertensive therapy on Ccr the reciprocal of the Pcr (1/Pcr). and their determinants (GFR, creatinine clearance from tubular secretion (CTScr), and creatinine excretion (UcrV) values) was assessed in patients participating in the Modification of Diet in Renal Disease (MDRD) Study. This study compared the effects of assignment to a low versus usual-protein diet and to a low versus usual-blood pressure goal on the decline in these measurements over 3 yr in 585 patients with baseline GFR of 25 to 55 mL/min per 1.73 m2 (Study A). This study also assessed correlations and associations of these measurements with each other and with protein intake, blood pressure, class of antihypertensive agents, and renal diagnosis in 840 patients with baseline GFR of 13 to 55 mL/min per 1.73 m2 (Studies A and B). In Study A, the estimated mean decline in GFR at 3 yr did not differ significantly between the low and usual-protein diet groups (-10.9 versus 12.1 mL/min). In contrast, CTScr declined more in the low-protein diet group ( 7.8 versus -3.6 mL/min, P < .05). Consequently, the low-protein diet group had a greater decline in Ccr (-17.6 versus -14.4 mL/min, P < .05). The low-protein diet group also had a greater decline in UcrV (-306 versus -92 mg/day, P < .05). The decline in UcrV was proportionately greater than the decline in CTScr hence the decline in 1/Pcr was less in the low-protein diet group (-0.091 versus -0.122 dl/mg, P < .05). Similarly, in Study A, there was no significant difference in the decline in GFR at 3 yr between the low and usual-blood pressure groups (-10.7 versus -12.3 mL/ min). However, there was a lesser decline in CTScr in the low blood pressure group (-4.7 versus -6.7 mL/ min, P < .05). Consequently, the decline in Ccr was less in the low blood pressure group (-14.2 versus -17.8 mL/min, P < .05). There was no significant difference in UcrV between the blood pressure groups (-192 versus -205 mg/day). Hence, the decline in 1/Pcr paralleled the decline in Ccr; it was less in the low blood pressure group (-0.091 versus 0.122 dL/mg, P < .05). In Studies A and B, correlations of rates of decline in Ccr and GFR were 0.64 and 0.79, respectively (P < 0.001). Correlations of rates of decline in 1/Pcr and GFR were 0.79 and 0.85, respectively (P < 0.001). In Studies A and B combined, baseline GFR, CTScr and UcrV correlated significantly with protein intake (r = 0.45, 0.47, and 0.36, respectively; P < 0.001), but not with blood pressure. Baseline CTScr was significantly lower in patients with polycystic kidney disease and tubulointerstitial diseases or urinary tract diseases, compared with glomerular and other diseases (P < 0.05). It was also lower in patients who were taking calcium channel blockers, compared with patients not taking these agents, and in patients not taking diuretics, compared with patients taking diuretics (P < 0.05). These results show that creatinine secretion and excretion are affected by protein intake. Creatinine secretion is also affected by antihypertensive therapy and renal diagnosis. In the MDRD Study, the low-protein diet reduced creatinine secretion and excretion, and the low blood pressure goal slowed the decline in creatinine secretion. These effects caused differences between the diet groups and between the blood pressure groups in Ccr and 1/Pcr that were not the result of differences in GFR. Studies assessing the effects of these interventions on the progression of renal disease should measure GFR in addition to Ccr and Pcr. PMID- 8724890 TI - Precision of glomerular filtration rate determinations for long-term slope calculations is improved by simultaneous infusion of 125I-iothalamate and 131I hippuran. AB - In studies on the progression of chronic renal failure the measurement of GFR must be very reliable. Sequential determination of GFR using the renal clearances of exogenous tracers such as inulin or iothalamate is the most accepted method. However, because of inaccuracies in urine collection, intratest variation, and thus intertest variation, of these clearances is considerable. This has a negative impact on the precision of long-term slope estimations. A previously described method of GFR determination on the basis of simultaneous infusion of 131I-hippuran and 125I-iothalamate corrects for inaccurate urine collection. To study whether this correction method improves the precision of the GFR slope measurement, this study analyzed longitudinal GFR data obtained in 71 patients with renal disease during a follow-up of 84 to 180 wk (477 renal function studies). All GFR were calculated by using both the standard renal clearance method and the correction method. The intratest and intertest coefficient of variation was significantly smaller for the correction method compared with the standard method (1.93 +/- 0.20 versus 8.48 +/- 1.66% P < 0.0005; and 2.88 +/- 0.32 versus 5.12 +/- 0.66%, P < 0.005, respectively). As a result, the precision of the GFR slope estimation was significantly better with the correction method compared with the standard method (error of the slope, 1.63 +/- 1.09 versus 2.35 +/- 2.36 mL/min per yr, P < 0.01). This improvement in precision of the slope by using the correction method reduces the necessary sample size needed to detect a GFR slope difference between interventions to about 30% of that needed when using the standard method. It is concluded that the precision of GFR measurements is improved by using correction for inaccurate urine collection with concomitant 131I-hippuran clearance. PMID- 8724891 TI - Both IgG- and C1q-receptors play a role in the enhanced binding of IgG complexes to human mesangial cells. AB - The presence of immunoglobulin G (IgG) in the mesangial area in kidneys of patients with different forms of glomerulonephritis suggests a role for IgG in the inflammatory process. This study investigates whether IgG is able to bind to cultured human mesangial cells (MC) in vitro. Incubation of MC with 125I aggregated IgG(125I-AIgG), as a model for immune complexes (IC), at 4 degrees C resulted in a time- and dose-dependent binding of 125I-AIgG to MC. The binding of 125I-AIgG to MC was inhibited by excess AIgG or Fc-fragments and not by F(ab')2 fragments or human serum albumin (HSA). Scatchard analysis revealed the presence of 2.8.10(6) receptors/cell with an affinity of 9.7.10(7) M-1. Incubation of MC with 125I-C1q resulted in a time- and dose-dependent binding of 125I-C1q to MC. The binding of 125I-C1q was inhibited by excess C1q or C1q talls and not by HSA. Scatchard analysis revealed the presence of 3.2.10(7) binding sites/cell with an affinity of 1.4.10(7) M-1. Immunoprecipitation of 125I-labeled MC membrane proteins with C1q or monoclonal antibodies directed against human C1q-R revealed a single 66 to 68 kd band under reducing conditions. Fluorescence-activated cell sorter analysis revealed an average of 60.1% +/- 5.4% of the cells positive with a mean channel of fluorescence of 592. A cooperative effect between C1q-R and Fc gamma-R in the binding of 125I-AIgG to MC, was assessed by incubation of 125I AIgG in the presence of increasing concentrations of C1q, C1q talls, or delta C1q. Only intact C1q showed a 6- to 11-fold enhancement in binding of 125I-AIgG to MC. These studies demonstrate the occurrence of C1q-R and Fc gamma-R on MC and indicate that binding of IC is enhanced after interaction of IC with C1q. PMID- 8724892 TI - Phosphodiesterase inhibitors correct resistance to natriuretic peptides in rats with Heymann Nephritis. AB - Experimental nephrotic syndrome is characterized by abnormal sodium metabolism, reflected in a blunted natriuretic response both to volume expansion and to infused atrial natriuretic peptide (ANP). The studies presented here examined the relationships among plasma ANP concentration and urinary sodium (VNaV) and cyclic GMP excretion (UcGMPV) in vivo, and the responsiveness of isolated glomeruil and inner medullary collecting duct (IMCD) cells to ANP and urodilatin (renal natriuretic peptide; RNP) in vitro in rats with Heymann nephritis, an immunologically mediated model of nephrotic syndrome. Nine to 14 days after Ip injection of anti-Fx1A antiserum, rats were proteinuric and had a blunted natriuretic response to intravenous infusion of isotonic saline (2% body weight, given over 5 min). Thirty min after the onset of the infusion, plasma ANP concentration was increased to the same extent in both normal and nephritic rats, compared with their respective hydropenic controls. Despite this increase, UcGMPV was significantly less in nephritic rats after the saline infusion. Accumulation of cGMP by isolated glomeruil and IMCD cells from nephritic rats after incubation with ANP and RNP was also significantly reduced, compared with normal rats. This difference was not related to differences in either density or affinity of renal ANP receptors, but was abolished when accumulation of cGMP was measured in the presence of 10(-3) M isobutylmethylxanthine or Zaprinast, two different inhibitors of cyclic nucleotide phosphodiesterases (PDE). Infusion of Zaprinast into one renal artery in nephritic rats normalized both the natriuretic response to volume expansion and the increase in UcGMPV from the infused, but not the contralateral, kidney. Furthermore, cGMP-PDE activity was increased in IMCD cell homogenates from nephritic compared with normal rats (388 +/- 32 versus 198 +/- 93 pmol/min per mg protein, P < 0.03). These results indicate that blunted volume expansion natriuresis accompanied by cellular resistance to ANP in vitro occurs in an immunologic model of renal injury. The resistance is not related to an alteration in ANP release or binding to its renal receptors, but is suppressed by PDE inhibitors and is associated with increased renal cGMP. PDE activity, thus suggesting that enhanced cGMP-PDE activity may account for resistance to the natriuretic actions of ANP observed in vivo. This defect may represent the intrinsic sodium transport abnormality linked to sodium retention in nephrotic syndrome. PMID- 8724893 TI - Colchicine interferes with L-selectin and leukocyte function-associated antigen-1 expression on human T lymphocytes and inhibits T cell activation. AB - Colchicine, which inhibits cell microtubule assembly by preventing polymerization of tubulin monomers, inhibits cell-mediated immune responses and promotes long term survival of major histocompatibility complex-incompatible renal allografts in rats. Here we evaluated the effect of blocking cell microtubule assembly by colchicine on T cell and endothelial cell adhesion receptors involved in transducing signals for T cell activation. By using immunofluorescence flow cytometry analysis, evidence is presented that colchicine, in a dose-dependent fashion, downregulated L-selectin and leukocyte function-associated antigen-1, but not CD2 and CD44 on the surface of naive human peripheral blood lymphocytes. This effect was confirmed in two subsets of T lymphocytes, namely, CD45RA- and CD45RO-positive cells. However, colchicine did not influence the rapid shedding of L-selectin from T lymphocytes exposed to activating stimuli. Colchicine inhibited expression of interleukin-2 receptor on activated T lymphocytes. This effect was observed when T lymphocytes were stimulated with both anti-CD3 and anti L-selectin monoclonal antibodies. Colchicine also inhibited lymphocyte function in vitro as documented by inhibition of the human mixed lymphocyte response in a dose-dependent fashion. Moreover, colchicine downregulated surface expression of intercellular adhesion molecule-1 and E-selectin on activated human umbilical vein endothelial cells. These results indicate that blocking cell microfubule assembly inhibits surface expression of adhesion molecules on T cells and endothelial cells, and provides insights into the complex mechanisms of the action of colchicine in vivo. PMID- 8724895 TI - The abnormal red-cell oxalate transport is a risk factor for idiopathic calcium nephrolithiasis: a prospective study. AB - An abnormal erythrocyte transmembrane oxalate flux was described in recurrent idiopathic calcium nephrolithiasis. To verify whether it might represent a risk marker of renal stone disease, two prospective studies were carried out. One hundred ninety patients with idiopathic calcium nephrolithiasis who were enrolled at their first episode of lithiasis during the period 1984 to 1986, form the basis of the first prospective study. The impact of erythrocyte oxalate transport anomaly, gender, familial occurrence of nephrolithiasis, hypercalciuria, hyperoxaluria, and hyperuricosuria on stone recurrence by both bivariate and multivariate analysis of frequencies was assessed. The predictive value of the erythrocyte anomaly for a patient's becoming a stone former was also assessed in five nephrolithiasis families. Recurrence occurred in 57.9% of patients; this was significantly associated with the erythrocyte anomaly, hyperoxaluria, and male gender. However, when using multivariate analysis, only gender and the erythrocyte anomaly were statistically significant and were independent predictors of recurrency. The probability of stone recurrency predicted by the logistic model ranged from 30.1% for women with normal erythrocyte oxalate transport, to 73.4% for men with the erythrocyte anomaly. The family follow-up showed that only subjects with the erythrocyte abnormality become renal stone formers in the 8-yr survey. By showing the predictive value of the erythrocyte oxalate anomaly for recurrent calcium nephrolithiasis, our findings support the notion that this anomaly is a risk factor in renal stone disease. PMID- 8724894 TI - Normal calcium oxalate crystal growth inhibition in severe calcium oxalate nephrolithiasis. AB - Urine from mammalian kidneys is regularly supersaturated with respect to calcium oxalate monohydrate, the most common solid phase in human nephrolithiasis, and also inhibits the nucleation, growth, and aggregation of calcium oxalate crystals. Nephrolithiasis is often associated with increased supersaturation, and it is assumed that this increase overbalances the inhibition effects, causing stones. However, some patients form stones in the absence of increased supersaturation, and in those patients, one might assume that reduced inhibition is the cause of their stones. This hypothesis was tested in 25 patients who formed at least ten stones each, yet lacked the usual metabolic abnormalities that increase supersaturation. Compared with 25 age- and sex-matched control subjects, urine supersaturation among the patients was not increased; this is an expected result of this study's selection criteria. Compared with the same age- and sex-matched control subjects, urine from the patients showed no evidence for reduced inhibition of calcium oxalate crystal growth, so low inhibition of growth did not contribute to pathogenesis of stones in our highly selected study population, despite their otherwise unexplained and active stone formation. These results do not support the hypothesis that growth inhibition defects are a cause of stone disease. PMID- 8724896 TI - Anomalous phospholipid n-6 polyunsaturated fatty acid composition in idiopathic calcium nephrolithiasis. AB - Anomalies in the erythrocyte transport of anions and cations have been described in idiopathic calcium oxalate nephrolithiasis and seem to play a pathogenetic role in this disease. In consideration of the hypothesis that the complex array of ion flux cell abnormalities is an epiphenomenon of an anomaly in the composition of cell membranes, this study investigated cell-membrane lipid composition. In idiopathic calcium oxalate renal stone formers, in which ion transport abnormalities were present, and in healthy control subjects, plasma and erythrocyte membrane lipid composition, the erythrocyte oxalate exchange, and Na/K/2Cl cotransport activity were evaluated. Furthermore, in stone formers, the effect of a 30-day fish-oil diet supplementation on plasma lipids, erythrocyte oxalate exchange, oxaluria, and calciuria was investigated. The effect of archidonic acid released by phospholipase A2 on anion-carrier phosphorylation and activity in erythrocytes was evaluated as well. Patients had a lower content of linoleic and higher concentration of archidonic acids in both plasma and erythrocyte membrane phospholipids, and an increased archidonic/linoleic acid ratio. The archidonic acid level correlated with the erythrocyte oxalate exchange and sodium cotransport activity. Fish-oil supplementation lowered calcium and oxalate urine excretion, and normalized the erythrocyte oxalate exchange. Phospholipase A2 increased the erythrocyte anion-carrier protein phosphorylation and the oxalate exchange. This study shows that idiopathic calcium nephrolithiasis in the patient group reported here is characterized by a systemic defect in phospholipid archidonic acid levels that might provide an answer to the link between genetic background, dietary habits, and renal lithiasis. PMID- 8724897 TI - Beneficial influence of ketanserin on autoregulation of blood flow in post ischemic kidneys. AB - The influence of ketanserin, a S2-serotonergic receptor blocker, on the impaired renal hemodynamics in a clamp model of renal ischemia in rats was investigated in this study. Serotonin-induced vasoconstriction of the renal vascular bed was augmented after ischemia. This constriction is blocked by ketanserin (0.05 mg/kg i.v. bolus, followed by 0.1 mg/kg per h infusion). The influence of the same ketanserin treatment on the response of RBF versus a stepwise lowering of the renal perfusion pressure was studied in post-ischemic kidneys with an established loss of autoregulation of RBF. An almost perfect restoration of the autoregulatory response was apparent after the S2-serotonergic antagonism. Despite this beneficial effect on renal hemodynamics, renal function, judged by measurement of GFR and urinary sodium excretion rate, was not influenced by an acute infusion of ketanserin in post-ischemic kidneys. It is suggested that serotonin plays a pivotal role in the suppression of autoregulation of RBF by a S2-serotonergic receptor-mediated vasoconstrictor effect in the post-ischemic kidney. It most likely masks the potential myogenic dilatory response of the smooth muscle cells in renal preglomerular microvasculature. Restoration of the renal autoregulatory capacity by S2-serotonergic receptor antagonism could be of clinical relevance in human post-ischemic acute renal failure. PMID- 8724898 TI - Abnormalities of heme biosynthesis in experimental acute renal failure. AB - Several abnormalities of porphyrin metabolism have been described in patients with end-stage renal failure. Because the heme biosynthetic pathway in acute renal failure has not been studied hitherto, an experimental model was therefore induced in 30 dogs by ligation and transection of both ureters. Forty-eight h after this procedure, anemia and uremia developed, erythrocyte aminolevulinate dehydratase activity decreased, and plasma porphyrins increased in these 30 dogs, whereas seven sham-operated animals did not exhibit any alteration of these parameters. Uremic plasma showed a capacity to inhibit aminolevulinate dehydratase activity (mean, 11.1 +/- 5.8%) when incubated in vitro with erythrocytes from healthy dogs. Such findings are similar to those reported in uremic patients on hemodialysis or on continuous ambulatory peritoneal dialysis. Twenty-three of the 30 animals underwent a hemodialysis session (180 min) 48 h after ureteral ligation, using a polyacrylonitrile membrane dialyzer. In addition to reducing serum creatinine and urea levels, this procedure significantly reduced plasma porphyrin values. However, the activity of erythrocyte aminolevulinate dehydratase and the plasma capacity to inhibit this enzyme were not modified after the hemodialysis session. This results described here show that some of the abnormalities of heme biosynthesis described in chronic renal failure are detected early in an experimental model of acute renal failure. This study also confirms that, although most plasma porphyrins circulate bound to proteins, hemodialysis may reduce levels of plasma porphyrins when a high permeability membrane is used. PMID- 8724899 TI - Thyroid hormone modulation of glucocorticoid-induced polycystic kidney disease. PMID- 8724900 TI - Recommendations for the initiation of dialysis. PMID- 8724901 TI - Low-calcium dialysate worsens secondary hyperparathyroidism. PMID- 8724902 TI - Pop-out in visual search requires practice. AB - It is not always easy to locate routing signs in the visually noisy environment of public buildings. Guidelines to ensure the conspicuity of an object in a complex field are not readily available. In two previous experiments, aiming to provide such guidelines for use in practice, the stimuli were simple simulations of scenes in public buildings, each containing a uniquely colored target (a routing sign) and a varying number of distractors (advertising signs). The results of these two experiments were opposite: in one experiment pop-out of the targets appeared as predicted by current theories of visual search, but in the other experiment performance significantly decreased when the number of distractors increased. Two new experiments are reported here to clarify this discrepancy. From the results it is concluded that pop-out of easy visual targets does not occur on the first few trials of a task. Subjects must have had sufficient practice for pop-out to develop. This finding implies that, contrary to what is common in experiments on visual search, short and heterogeneous series of stimuli should be used to ensure the practical relevance of the results for applications such as sign posting. PMID- 8724903 TI - Exercise dependence: the relationship between two measures. AB - A volunteer sample of 158 aerobics instructors, 46 competitors, and 102 general participants completed 2 assessments of exercise dependence, the Negative Addiction Scale and the Commitment to Running Scale. Pearson rs showed scores on the measures related significantly for the total sample (.33), the participants (.35), and the instructors (.36), but the correlation of .23 for scores of competitors was not significant. Although scores on the two scales were significantly correlated, each test accounted for less than 11% of the variance for the total sample. While there is some commonality, these instruments largely measure different qualities and care should be taken when comparing "exercise dependence" across samples if assessed by different tests. PMID- 8724904 TI - Internal consistency of boredom proneness scale for a sample of Pakistani students. AB - This is a short report of a new scale for the measurement of boredom for college students in Pakistan. An innovative technique was used. Internal consistency was .82 and convergent validity .30. Other estimates of reliability and validity are planned. PMID- 8724905 TI - Discriminant effectiveness of psychological state measures in predicting selection during field hockey trials. AB - Field hockey players (N = 128) completed the Competitive State Anxiety Inventory 2 and the Profile of Mood States about 45 min. before a British Universities trial. Single-factor multivariate analysis of variance indicated no significant differences between selected and nonselected players for any preperformance mood or anxiety measure. Discriminant function analysis showed that 74 participants (57.81%) could be correctly classified as selected or nonselected players on the basis of preperformance mood scores. This figure rose to 83 participants (64.84%) when scores on the anxiety subscales were also included in the discriminant function analysis. Anxiety scores alone discriminated 71 participants (55.47%). These results concur with earlier proposals of Terry that psychological state measures decline in predictive effectiveness in long duration, open skill team sports. PMID- 8724906 TI - Human memory for odour following monorhinal presentation with and without verbal elaboration. AB - With 14 target odours, 7 via each nostril, 20 subjects' correct recognition was lower for the right nostril than for the left with verbal elaboration, but nonsignificant without verbal elaboration. PMID- 8724907 TI - Visual recognition training of older adults with speech spectrograms. AB - 10 adults' performances on visual training through recognition of speech spectrograms were examined. All subjects completed the training within eight 1 hr. sessions. Success and retention of training were also evident in the subjects' performances on two posttests. PMID- 8724908 TI - Pattern of efficacy of a musical mnemonic on recall of familiar words over several presentations. AB - It has been demonstrated that musical accompaniment may aid recall of words. This study examined the mnemonic effect of music on recall of verbal material which contained no unfamiliar words over three presentations. 20 participants (average age 21.9 yr.) were randomly allocated to one of two experimental conditions and heard the to-be-remembered lyrics either being sung or read aloud without musical accompaniment. Analysis of both total number of words correctly recalled and the extent of chunking of recalled material showed better over-all recall in the song condition with evidence of greater chunking of material. PMID- 8724909 TI - Inhibition of the soleus H-reflex during dorsiflexion is dependent on individual differences in maximal soleus H-reflex as a test reflex. AB - The quantitative differences among individuals in the natural reciprocal inhibition of the soleus H-reflex during dorsiflexion were examined, in conjunction with the maximal H-reflex as the test reflex size in each individual. Maximal H-reflex was expressed relative to the maximal M-response (H(max)) when compared among individuals. Analysis showed that with increases in H(max) at rest in each individual, the inhibitory effect was first enhanced, then reached a peak, and was finally alleviated. This pattern was similar to the intraindividual pattern of the inhibitory effect induced by specific conditioning stimulus as a function of the test reflex size. PMID- 8724910 TI - Perceived exertion during physical exercise among individuals high and low in fitness. AB - This study was conducted to investigate the influence of fitness level and gradually increased amounts of exercise on individuals' ratings of perceived exertion (RPE). 20 men served as paid subjects. They were divided into groups of high ( > 56 ml/kg/min.) and low fitness ( < 46 ml/kg/min.) according to their maximal oxygen uptake (VO2 (max)). Participants were required to pedal on a cycle ergometer at a progressively increased workload (every 10 min.) corresponding to 40, 50, 60, 70, and 80% of individual VO2 (max) values. Heart rates, RPE, and core temperatures were recorded every 5th minute after the initiation of exercising at a specific workload. Analysis indicated that, when controlling for VO2 (max) values, elevations of heart rate and core temperature were not affected by fitness. However, highly fit individuals perceived themselves under less exertion than did the group low in fitness. Correlations showed that, taking into consideration fitness, there is a stronger relationship between RPE and heart rate and RPE and core temperature for the highly fit individuals than for the less fit. PMID- 8724911 TI - Behavior modification with brain-injured children: a brief note on directions for research. AB - Research on behavior modification needed with brain-injured children is briefly noted. Previous studies indicate that the procedures of behavior modification work well with selected brain-injured children. The designs of most studies, however, have used small Ns or a single subject to evaluate interventions with brain-injured children. Some suggestions for research needed with this population are provided. PMID- 8724912 TI - EEG correlates of enhanced spatial performance following exposure to music. AB - Previous research has shown that exposure to classical music can influence performance on a spatial task. The present study investigated EEG correlates of this enhanced performance effect, 4 female and 4 male undergraduates completed two equivalent spatial tests, one following a control procedure and one following the presentation of Mozart's Sonata for Two Pianos in D Major. EEG was recorded during a baseline and two task-performance periods. Test performance and EEG recordings were analyzed, and correlations were generated between task performance and EEG variables (average spectral power and peak frequency within 5 frequency ranges). Performance improved significantly following the presentation of the music. EEG analysis indicated 6 reliable correlations out of 40 calculated between differential EEG variables and changes in performance. Ten reliable correlations out of 120 were also found between changes in performance and nondifferential EEG variables across baseline, control, and experimental conditions. PMID- 8724913 TI - Walking patterns and finger rhythm of older adults. AB - Walking patterns and rhythmic movement of the fingers were examined in a total of 1,134 male and female community residents 65 years of age and over. Walking patterns were characterized according to the ratio of step length divided by step rate (cadence), called the Walk Ratio, during level walking at preferred and maximum speeds. The walking pattern tended to change according to age; older subjects walked with shorter steps (smaller Walk Ratio). Rhythmic movement was examined using the finger-tapping test in time to the sound of a 4-Hz metronome. Hastened tapping or finger festination, in which the subject tapped faster than requested (constant error of 3 msec. and over in the intertap interval), was characteristic of aging; 16.8% of the subjects exhibited finger festination and the occurrence increased with age, especially among those in their eighties (29.3%). Finger festination was accompanied by walking patterns with an increased step rate, or a smaller Walk Ratio. These characteristics of aging were discussed as similar to extrapyramidal symptoms of walking and rhythm production in patients with Parkinson's disease. PMID- 8724914 TI - Relationship between motoneuron pool excitability and parameters of timing and force in an isometric ankle-extension movement. AB - The changes in peak facilitation and slope of facilitation of the H-reflex were compared to the parameters of an isometric plantarflexion task. The amplitude of the reflex was significantly greater for reflexes elicited within 50 msec. of the onset of soleus muscle activity. Neither peak amplitude of the reflex nor slope correlated with rate of rise of isometric force; however, they did show a modest relationship between premotor time and reaction time. PMID- 8724915 TI - Analysis of position of trial sequence and type of diluent on the detection threshold for phenyl ethyl alcohol using a single staircase method. AB - Although detection thresholds for odors are commonly measured in academic and medical settings, the influences of procedural factors on threshold values are poorly understood. The present study evaluated the influences of (i) trial sequence position and (ii) diluent type on the threshold value for the rose-like odorant phenyl ethyl alcohol. In Exp. 1, detection thresholds were measured in 24 subjects on two occasions in which different diluents were used in the concentration series, propylene glycol and light mineral oil. The thresholds were estimated using a 7-reversal initially ascending single-staircase procedure. Threshold values were significantly influenced by the type of diluent (lower for mineral oil) and trial sequence (lower for later threshold reversals). In Exp. 2, 24 subjects were administered a staircase threshold test which continued through 15 staircase reversals. Continued testing resulted in a significant lowering of the threshold measure. These findings demonstrate the importance of both diluent and test length on detection threshold values measured by a single staircase procedure and emphasize the need for standardization of procedures for threshold testing. PMID- 8724916 TI - Effects of stimulus duration, temporal delay, and complexity on the judgments of dot location. AB - The present study investigated how information on location was processed at an early stage of information processing by the task of recognizing dots presented briefly and followed by a masking stimulus. 8 subjects were asked on 1528 trials to recognize the positions of dots presented on a circumference. Effects of number of dots (1, 2, and 3), duration of presentation (36, 56, 200, and 500 msec.), and time delay (interval between the offset of display and the onset of recognition stimulus, 100 and 500 msec.) were examined. Analyses showed that the percentage of correct recognition increased with the duration of presentation and that the effect of duration decreased with the number of dots. Meanwhile, no significant effect was found for time delay. Consequently, with increased duration, the information on location is transferred to short-term visual memory. However, the ratio of transfer from iconic storage to short-term visual memory with duration varied with the complexity of visual stimulus. PMID- 8724917 TI - Visuospatial/vocabulary differences in boys and girls and a potential age dependent drift in vocabulary proficiency. AB - A total of 188 monolingual (English) boys and girls whose ages ranged from 6 years through 14 years were administered the Vocabulary and Block Design subtests of the Wechsler Intelligence Scale for Children. Although the boys (n = 85) displayed significantly greater relative strengths for the visuospatial task than the girls (n = 103), the effect was due primarily to the boys' less proficient Vocabulary scores. The detection of a significant linear decrease (r = -.32) in standardized scores for Vocabulary but not for Block Design over this age span was not expected. The drift over 9 grades was equivalent to more than one standard deviation. Possible sources of interference such as exposure to the same second language that occupied 6% to 12% of the instructional time are suggested. PMID- 8724919 TI - Association of prelesion sports participation and involvement in wheelchair sports following spinal cord injury. AB - 116 individuals with spinal cord injury were surveyed to investigate the relationship between prelesion sports participation and involvement in wheelchair sports following injury. While 89 reported they had been involved in sport prior to their injury, only 38 (43%) of these were involved since injury. 27 respondents said that they had not participated in sport preinjury; however, 13 (48%) of these reported that they had become involved since injury. Analysis by chi 2 showed that, for this sample at least, the tendency for an individual with spinal cord injury to be involved in wheelchair sports was not related to preinjury history of sports involvement. PMID- 8724918 TI - Use of the long vs short form of the speech sounds perception test in a school age population. AB - A sample of 645 school-age children, ages 9 to 14 years, were administered independent forms of the Speech Sounds Perception Test for older children. 328 subjects took the complete test (SSPT-60) and 317 were administered only the first 30 items (SSPT-30). Comparison of mean errors indicated that significantly more errors were made by subjects given the SSPT-30. Decisions regarding the use of each form for school-age children are discussed. PMID- 8724920 TI - Percept-genetic codings of defense in chronic schizophrenia. AB - Hitherto available studies on the percept-genetic defensive organization of Schizophrenia have not distinguished between acute and chronic stages of the disorder. The present research with the Defense Mechanism Test included 30 chronic inpatients with several years of hospitalization and with acceptable perceptual thresholds. Compared with 30 sex- and age-matched nonschizophrenic psychiatric control patients, schizophrenics resorted significantly more often to (a) regression, (b) disappearance of the peripheral figure, (c) introjection (wrong sex attribution to the hero), and (d) significantly less often to the most mature variants of repression. In a further comparison of a subgroup of 16 women schizophrenic patients and a matched group of melancholic inpatients, the findings on regression, introjection, and repression were replicated. PMID- 8724921 TI - Effect of instruction on perceived physical ability and exercise adherence. PMID- 8724922 TI - Effect of carbohydrate substrate availability on ratings of perceived exertion during prolonged exercise of moderate intensity. AB - This investigation examined the effect of carbohydrate substrate availability on ratings of perceived exertion (RPE) during cycling at moderate intensity to exhaustion and the relation between submaximal endurance performance and RPE obtained following 2 hr. of cycling at moderate intensity. Seven male cyclists performed two exercise trials at power output corresponding to 70% of their peak oxygen uptake until exhaustion. Subjects ingested either a 6% glucose/sucrose solution at the rate of 0.6 g.kg-1 (Body Weight).hr.-1 or an equal volume of artificially flavored placebo every 20 min. throughout the exercise trials. RPE for the legs, chest, and over-all body, and oxygen consumption, expired ventilation, carbohydrate oxidation rate, and blood concentrations of glucose, glycerol, and lactate were measured every 20 min. throughout exercise and at exercise termination. Statistical analysis of these dependent variables indicates that (1) an exercise-induced decrease in blood-borne carbohydrate substrate intensifies leg and over-all perceptions of exertion during the later stages of prolonged cycling at 70% VO2peak. (2) Factors other than availability of blood borne carbohydrate substrate may influence perceptual intensity at exhaustion. (3) Ratings of perceived exertion for the legs and over-all body obtained after 2 hr. of cycling at moderate intensity may be useful in predicting submaximal endurance performance. PMID- 8724923 TI - Augmented feedback for enhanced skill acquisition in individuals with traumatic brain injury. AB - Coincident timing by individuals who exhibit traumatic brain injury was measured under conditions of no knowledge of results (no KR; n = 12), KR on every trial (n = 14), summary KR (n = 13), and average KR (n = 12). Following acquisition trials, groups performed immediate and longer retention trials without KR. Absolute constant error and variable error, analyzed in separate repeated measures analyses of variance, indicated that during acquisition trials subjects receiving KR on every trial were the most accurate and the most consistent in their responses; however, subjects in groups receiving summary and average KR were the most accurate during immediate retention, with the group receiving summary KR being the most accurate during longer retention. PMID- 8724924 TI - Influences of aging, gender, and handedness on motor performance of upper and lower extremities. AB - The purpose of this study was to collect reference data on different aspects of performance, including reaction time, speed of movement, tapping speed, and coordination of hands and feet using Human Performance Measurement/Basic Elements of Performance equipment and reports of reference data findings. Also, the reliability of the measurements is presented. 200 healthy, randomly selected subjects (100 men, 100 women; aged 21-70 years) were categorized by gender and by age decade into ten groups. The test battery consisted of six tests for both hands and feet. In general, the performance decreased clearly after 50 years in both genders. There were statistically significant differences between hands and feet, dominant and non-dominant sides, age groups, and number of choices, and especially between men and women. PMID- 8724925 TI - Reactivity in extroverts and introverts. AB - The response times of 17 introverts and 24 extroverts to red and blue stimulus lights were compared. While no differences were found between the groups, introverts responded faster to the presentation of the red stimulus light than the blue one. PMID- 8724926 TI - Effects of bandwidth feedback on delayed retention of a movement timing task. AB - The effectiveness of broad and narrow bandwidth feedback for learning a 2-segment timing task was tested over short-term (10-min.) and longer-term (2-day) retention intervals. The main finding was that bandwidth feedback groups deteriorated in accuracy across the retention tests, raising questions about the effectiveness of bandwidth feedback for longer-term learning. PMID- 8724927 TI - An initial evaluation of work fatigue and circadian changes as assessed by multiplate posturography. AB - The sensitivity of postural responses to the effects of work fatigue and circadian changes was explored in a pilot study using a specific method of multiplate posturography based on the differential assessment of vertical pressure on four separate platforms for each heel and set of toes of each foot, respectively 8 normal subjects, M.D.s, were given six posturographic examinations immediately before and after three 8-hr. work shifts in the emergency room of a major hospital in Tel-Aviv. 3 posturographic measures (stability, spectral power of postural sway at low frequency of 0.1-0.25 Hz, and unilateral weakening of heel-toe synchronisation) were significantly affected by work fatigue whilst also showing interaction with circadian rhythm. 2 additional measures (power of sway at high frequency of 1.00-3.00 Hz and dysharmonic distribution of weight over the four platforms) were not related to workload but showed significant circadian changes. These effects appeared only on positions involving restricted visual and somatosensory feedback causing vestibular stress. Results justify the application of multiplate posturography as an ancillary tool in measuring objectively the effects of fatigue and circadian changes as well as the interaction between endogenous chronobiological processes and their external conditioning factors (Zeitgebers). Pragmatic implications of the findings in the context of industrial medicine and interdisciplinary efforts to prevent road and air accidents are discussed. PMID- 8724928 TI - An examination of error reductions in consonant recognition when vision and hearing are combined. AB - Errors in judging by 9 men and 9 women the three acoustic cues for consonant recognition, those of voice, place of articulation, and manner of articulation, were markedly reduced by 70% or more when vision was added to hearing in a word in-noise discrimination task. The extent of error reduction is surprising especially for the least-visible voice cue. PMID- 8724929 TI - Null effects of facial asymmetry on intrafacial speechreading performance in men and women. AB - Intrafacial differences in speechreading performance were studied measuring 24 subjects' visual recognition of key words within sentences when presented either a left-left-(LL) or right-right-(RR)-sided face of a male speaker with moderate facial asyrnmetry. No significant differences were found as a function of facial side or gender (ns = 12). PMID- 8724930 TI - Effects of hypnosis on the fundamental frequency of voice in different emotional imitations. AB - The modifications of human vocal fundamental frequency (fa) were studied in six emotional imitations of a sequence during normal and hypnotic conditions. In hypnosis the fundamental frequency was significantly lower than in the normal condition; in particular, in this altered state of consciousness two emotional imitations (angry and crying conditions) were significantly different with respect to the normal condition. PMID- 8724931 TI - Language production in younger and older adults. AB - Only six of 74 linguistic variables studied were affected by age when younger and older subjects were compared. Three of these variables related to pragmatics, two to semantics, and one to syntax. No differences were noted for the parameter of phonology. PMID- 8724932 TI - Temporal consistency and stability of three scales for measuring stress in adults. AB - We investigated the 30-min. temporal consistency and temporal stability of three focal scales for measuring stress: the Cognitive Anxiety Scale-State, the Focusing Inventory Scale-State, and the Somatic Symptom Scale-State. These scales are applicable for individually measured reduction in stress in research on outcome with older adolescents and adults. All possess excellent internal consistency. All manifested significant temporal consistency and good to excellent temporal stability over 30 min. in a nonclinical sample of 60 undergraduates. Small significant differences in means on the Cognitive Anxiety Scale-State and the Somatic Symptom Scale-State were noted. The phenomenological states associated with stress were stable and consistent over 30 min., the optimal time interval used in research on stress. PMID- 8724933 TI - Improvements in horizontality performance as a function of type of training. AB - 90 undergraduates with low spatial perceptual ability were assigned to one of three groups of control, learner-guided instructions, and tutor-guided instructions. The groups differed in amount and type of spatial training administered in a laboratory setting. Participants completed horizontality and vertically spatial perceptual tasks immediately after pretesting and again one month later. Both groups given training showed more accurate performance at posttest than at pretest. In addition, the learner-guided instruction group outperformed the tutor-guided instruction group and control group at posttest and on novel but related tasks. Gender differences in posttest performance favoring males (n = 35) were obtained in the tutor-guided performance group and control group but not in the learner-guided instruction group. The number of hints the latter needed during training was predictive of posttest performance. Implications for assessment and remediation of undergraduates' spatial perceptual abilities are discussed. PMID- 8724934 TI - A looming-recession threshold. AB - When the origin of magnification-minification of an outline rectangle had a horizontal locus which exceeded one-fourth of the rectangle's horizontal dimension, 16 observers of 21 reported apparent depth characteristic of looming and recession. PMID- 8724935 TI - Benussi effect as an integration of disparity by motion. AB - It was hypothesized that the apparent depth of the Benussi effect could be elicited using stimuli corresponding to retinally disparate images. When such stimuli were presented rapidly and successively in a manner approximating apparent motion, 21 of 22 observers reported apparent depth. PMID- 8724936 TI - A new method for examining the judgment of bigness. AB - 80 children (40 three-yr.-olds, 23 boys and 17 girls, and 40 five-yr.-olds, 22 boys and 18 girls) were presented pairs of quadrangles (square vs rectangle) and triangles (two isosceles triangles) and asked to select the bigger one. The pairs were varied on the area ratio of paired stimuli and magnitude in differences between height and width. Although 3-yr.-olds tended to rely on area in making their judgments and also occasionally relied on height, almost all 5-yr.-olds relied on height. These results were discussed in relation to the tasks used in the previous studies, the basis in judgment of bigness, and the Japanese word for "big." PMID- 8724937 TI - Attempts to simulate the association between geomagnetic activity and spontaneous seizures in rats using experimentally generated magnetic fields. AB - In three separate experiments, chronic epileptic male rats (ns = 24, 24, and 4) were exposed to experimental magnetic fields whose intensities were either constant or varied. The ripple frequency was always 7 Hz. Only exposure to a field with a constant intensity (700 nT) appeared to inhibit occurrence of the seizures. Although exposure to a field configuration whose strength increased and decreased by fixed increments during the hour before feeding did not affect the incidence of seizures, exposure to these fields for 5 min, only once per hour (increments of 15 to 20 nT for 30 sec. to a maximum of 70 nT) during the previous night between midnight and 0800 hr. increased the proportion of seizures following the presentation of food. An explanation is given for the persistent antithetical effects of experimental magnetic fields and geomagnetic activity upon seizure phenomena. PMID- 8724938 TI - Effects of background stimulation upon eye-movement information. AB - To investigate the effects of background stimulation upon eye-movement information (EMI), the perceived deceleration of the target motion during pursuit eye movement (Aubert-Fleishl paradox) was analyzed. In the experiment, a striped pattern was used as a background stimulus with various brightness contrasts and spatial frequencies for serially manipulating the attributions of the background stimulus. Analysis showed that the retinal-image motion of the background stimulus (optic flow) affected eye-movement information and that the effects of optic flow became stronger when high contrast and low spatial frequency stripes were presented as the background stimulus. In conclusion, optic flow is one source of eye-movement information in determining real object motion, and the effectiveness of optic flow depends on the attributes of the background stimulus. PMID- 8724939 TI - On-line control of a speeded motor response. AB - Evidence for a point of no return in a motor act has been reported to occur very late in its preparation and even to the point at which the response is executed. We report a qualitative example from electromyographic (EMG) data of an elbow extension movement from 1 of 12 adults which suggests that the response can be stopped at any time up to its production, from which we conclude that the response is subject to on-line control at all times. PMID- 8724940 TI - Repeated verbal interruptions during exposure to complex transcerebral magnetic fields elicit irritability: implications for opiate effects. AB - Men (n = 17) and women (n = 18) listened to a brief narrative and then were exposed for 30 min. to a control condition or to a weak (1 microTesla) complex magnetic field applied over the left hemisphere, the right hemisphere, or bilaterally. The subjects were interrupted intermittently to report their thoughts during this period. Because the wave structure of the field had been shown to elevate nociceptive thresholds and to simulate the effects of morphine in rats, we predicted that the group who received the bilateral stimulation should report greater irritability because of their disrupted pleasant experiences. The results supported this hypothesis. PMID- 8724941 TI - Performance differences as a function of stimulus-response compatibility with rapid serial visual presentation and spatially distributed presentation styles. AB - A means of presenting information temporally has been developed from the basic research on reading. Temporal displays allowed for faster processing speeds by reducing the number of saccades normally required to process spatially distributed information; however, a potential disadvantage to using temporal displays was an increased rate of error. Payne and Lang reported that a tradeoff between speed and accuracy is sometimes the result of using temporally distributed displays. As they used an alphabetic coding scheme which may not have facilitated the most optimal stimulus-response mapping, the current experiment tested the effects of a directional coding scheme on a temporally distributed display with 24 subjects. Analysis indicated the coding scheme did improve processing speed on the temporal display, but the error rate for the temporal display was higher than with the alphabetic coding scheme. Therefore, the limitation of temporal displays in a tradeoff between speed and accuracy persisted using the directional coding scheme. PMID- 8724942 TI - Children's perceptions of their current and ideal body sizes and body mass index. AB - 244 Australian schoolchildren aged between 8 and 12 years indicated their current and ideal body sizes by means of Collins' pictorial figures. Children's height and weight were also measured. Body-mass indices were calculated from these data, and percentile ranks estimated according to international reference data. Consistent with Collins' findings, 39% of girls and 26% of boys wanted to be thinner than they perceived themselves to be; however, this desire was strongly related to actual body size so the percentages were very different for weight categories established on the basis of Body Mass Index. In the overweight quartile, 76% of girls and 56% of boys wanted to be thinner, whereas in the underweight quartile only 10% of girls and no boys wanted to be thinner. The response of overweight children may be sensible, but the desire of some underweight girls to be even thinner is of concern. Some recent literature suggests that underweight individuals tend to overestimate their body sizes. About a half of our underweight children slightly overestimated their body sizes but only one child overestimated grossly. PMID- 8724943 TI - Scores on sensation seeking as a predictor of sexual activities among homosexuals. AB - Sensation seeking scores were positively correlated with the variety of homosexual activities and the number of homosexual partners within a gay population of 100. However, sensation seeking scores were not correlated with risky sexual practices. PMID- 8724944 TI - Development of cursive handwriting. AB - The efficiency of two different instructional methods for improving the growth of handwriting quality in the upper grades of primary school was investigated in field experiments using a control-group design. No improvement in mean quality was found for a group of 38 children using a method based on copying exercises. A substantial improvement in mean quality, corresponding to about 1.4 SD, was found for 23 children using a method based on reintroduction of the letter forms explaining each form visually and verbally. It was concluded that the bottleneck in the development of handwriting quality in primary school is a mismatch between the instructional methods used and the perceptual ability of the children. PMID- 8724945 TI - Spatial and temporal locations in child Jordanian Arabic. AB - The purpose of this research was to evaluate the influence of cognitive development on the acquisition of the spatial and temporal systems in Jordanian Arabic. 60 Jordanian children 2 to 6 years old received a comprehension test based on a 1991 sentence-picture matching task of Weist, wherein each problem contained a minimal morphological contrast. These contrasts were either spatial, e.g., 'in/on', or temporal, e.g., past/future tense. Further, the contrasts required either a single referent object or event, e.g., 'in/on' and past/future tense, or they required two or more referent objects or events, e.g., 'between' and 'before/after'. Firstly, significant change across age groups was noted. Secondly, problems which required two referent objects or events were more difficult than those requiring one referent object or event. Finally, spatial contrasts were easier than temporal ones. The findings were related to the general issue of the interaction of language and thought during the acquisition of language. PMID- 8724946 TI - Geophysical variables and behavior: LXXX. Periodicities and energetic characteristics of a strobe-light luminosity during a geomagnetic storm. AB - A videotape of a "strobe-light" luminosity that occurred in northern Ontario for several minutes during the evening of 6 October, 1994, at the time of a geomagnetic storm, was analyzed frame by frame. Brightness of the flashes decreased over the time of the observation. The interflicker intervals displayed phase-shifted periodicities that would be compatible with a rotating electromagnetic dipole and with the observations of the witnesses. The time of the occurrence and the dynamics of the luminosity were consistent with the predictions of the tectonic strain hypothesis. PMID- 8724947 TI - Assessing functional status of elderly adults via microcomputer. AB - Geriatric adults (at least 65 years of age) completed the Functional Status Questionnaire twice, first using the original paper-and-pencil format and again using one of six computer-based interactive methods. Scoring via original and computer-based methods was identical. Satisfaction with computers was high. Of the interactive methods, touchscreen required least training and questionnaire completion time and was rated most useful. PMID- 8724948 TI - Motivation to participate, exercise affect, and outcome behaviors toward physical activity. AB - Recently researchers have focused attention on understanding the relationship between motivation to participate and affective states associated with adherence to exercise and psychological outcomes of participation. It was hypothesized that intrinsic motivation toward an activity will heighten the associated positive affect, thereby leading to increased participation and higher perceived competence and satisfaction. The presently devised models tested the above hypothesis with 130 college students enrolled in psychology classes. They were asked to complete the Motivation for Physical Activity Measure, the Sport Enjoyment Questionnaire, and general demographic questions assessing adherence to exercise and perceived competence and satisfaction. Partial support for these path models was shown as intrinsic motivation was a predictor of affect and perceived competence and satisfaction, while extrinsic motivation predicted adherence for men only. These results provide a model to enhance the understanding of exercise-related behavior in the general population and the study of the relationship of motivation with affect toward exercise. PMID- 8724949 TI - Errors on cues for consonant identification on auditory and visual word recognition tasks. AB - For 9 women and 9 men undergraduates the proportion of voice, place of articulation, and manner of articulation errors were significantly different on an auditory word-in-noise task but not on a visual word-discrimination task. Results are most surprising for the visual condition, since voice cues are considered the least visible of the three cues for identification of consonants. PMID- 8724950 TI - Myelinated nerve fibers and retinal vascular abnormalities. AB - BACKGROUND/PURPOSE: Abnormal retinal vessels may develop in a region of myelinated nerve fibers, and these vessels may cause vitreous hemorrhages. METHODS: The clinical histories of seven patients with retinovascular abnormalities in a patch of myelinated nerve fibers are presented. None of the reported patients had other evidence of systemic disease. The cases were traced by a multicentric retrospective study. RESULTS: Retinal vascular abnormalities ranged from mild telangiectasis to frank neovascularization, with or without obstruction of the capillary network and signs of branch artery and vein occlusion. Age at diagnosis ranged from 15 to 68 years. Vitreous hemorrhages occurred in the four youngest patients and occurred at 15, 27, 27, and 43 years of age. Laser photocoagulation was applied in three patients and vitrectomy was performed in one. CONCLUSION: The authors' findings suggest that the abnormal structure of the myelinated nerve fibers and the thickened nerve fiber layer of the affected portions of retina may play a role in the onset of retinal vascular abnormalities and eventually cause telangiectasis, branch artery and vein occlusion, neovascularization, and vitreous hemorrhages. This suggestion is based on the absence of other causes of neovascularization or vitreous hemorrhage in all seven patients, and on the relatively young age of four of the patients with this association. PMID- 8724951 TI - Indocyanine green angiographic aspects of multiple evanescent white dot syndrome. AB - PURPOSE: The authors investigated the indocyanine green angiography findings of multiple evanescent white dot syndrome (MEWDS). METHODS: Four patients with MEWDS underwent examination by indocyanine green angiography, conventional ophthalmoscopy, and fluorescein angiography. RESULTS: Fundus examination showed multiple white dots in the retinal pigment epithelium of the unilateral eye of each patient. Fluorescein angiography demonstrated early hyperfluorescence corresponding to the white dots. In the early phase, indocyanine green angiography showed no abnormal signs in the large choroidal vessels, but in the late phase, hypofluorescent lesions appeared, corresponding to the white dots. The hypofluorescent dots were clustered in the posterior pole and sporadic in the peripheral region, appearing to radiate away from the optic disc or fovea. The hypofluorescent dots disappeared at the recovery stage. CONCLUSIONS: Previous fluorescein angiographic and electrophysiologic studies have demonstrated the involvement of the retinal pigment epithelium (RPE) and photoreceptors in MEWDS: The current findings on indocyanine green angiography suggest that MEWDS affects the choriocapillaris or precapillary arterioles as well as the RPE and photoreceptors, and that the lesions spread to the midperipheral region, centering on the optic disc or fovea. PMID- 8724952 TI - Idiopathic polypoidal choroidal vasculopathy associated with retinal arterial macroaneurysm and hypertensive retinopathy. AB - BACKGROUND: Idiopathic polypoidal choroidal vasculopathy (IPCV), the posterior uveal bleeding syndrome, and multiple recurrent serosanguineous pigment epithelial detachments in black women are a heterogeneous group of related entities seen in elderly hypertensive females. Similar associations are known for retinal arterial macroaneurysm (RAM). METHODS: This report documents the clinical, fluorescein, and indocyanine green videoangiographic findings of two elderly black female hypertensive patients who presented with IPCV, RAM, and hypertensive retinopathy. RESULTS: Clinical, fluorescein, and indocyanine green videoangiographic analysis of these two patients revealed hypertensive retinopathy, RAM, and anomalous polypoidal vascular networks associated with focal "bead-string" constrictions, terminal vascular loops, and polypoidal and pinpoint choroidal nodules. CONCLUSION: Certain subgroups of IPCV and related entities and RAM may be pathophysiologically related as analogous hypertensive insults to the choroidal and retinal vascular beds. PMID- 8724953 TI - Decision-making in the treatment of subfoveal neovascularization in age-related macular degeneration. An analysis from the patient's perspective. AB - BACKGROUND: The clinical management of subfoveal neovascularization in age related macular degeneration remains controversial. Large multicenter clinical trials recommend laser photocoagulation for certain subfoveal membranes, but many ophthalmologists are reluctant to treat patients with such lesions because a substantial initial visual loss may be induced by the treatment itself. Despite these controversies in management, the attitude of patients toward treatment of these lesions has not been assessed systematically. METHODS: In this article, a newly designed survey containing five hypothetical clinical scenarios was given to 68 patients with age-related macular degeneration to measure their relative preferences for the potential long-term benefits of laser photocoagulation compared with the immediate visual loss induced by laser therapy. RESULTS: This pilot study reveals a strong preference by these patients for laser intervention, despite the risk of immediate visual loss. On all five questions, the laser treatment outcome was preferred over the natural course of the disease. CONCLUSION: Ophthalmologists, when making clinical recommendations to patients with subfoveal exudative macular degeneration, may wish to consider the results of this survey, which suggest a substantial patient preference for laser treatment versus no intervention. PMID- 8724954 TI - Cytomegalovirus retinitis and optic neuritis in a child with severe combined immunodeficiency syndrome. AB - BACKGROUND: Although cytomegalovirus (CMV) infection of the retina and brain is common in patients with acquired immunodeficiency syndrome (AIDS), it is exceedingly rare in patients with immunodeficiencies due to other causes. This is the first report on ocular and cerebral histopathology of disseminated CMV in a child with severe combined immunodeficiency syndrome (SCID). METHODS: The authors examined by routine histopathologic methods the eyes of a 2-year-old white boy with SCID and bilateral CMV retinitis who died after failure of a third attempt at allogeneic bone marrow transplantation (BMT). RESULTS: Cytomegalovirus inclusions were found in the necrotic retinal remnants, in the hyperplastic and scarred retinal pigment epithelium, and bilaterally in the optic nerves. There were infiltrates of macrophages in response to the infection or the infused silicon, but no lymphoid infiltrates. Cytomegalovirus inclusions also were found in brain tissue. CONCLUSION: The histologic features resembled those of CMV retinitis and optic neuritis in AIDS. PMID- 8724955 TI - Mycobacterium fortuitum endophthalmitis. AB - PURPOSE: The authors report the first known case of mycobacterium fortuitum endophthalmitis. METHOD: The authors use the documentation of clinical course. RESULTS: A postoperative cataract patient received a diagnosis of mycobacterium fortuitum endophthalmitis 1 month after uncomplicated cataract surgery. The endophthalmitis responded to intraocular amikacin. CONCLUSIONS: When mycobacteria are recovered from patients with endophthalmitis, amikacin should be included in the therapeutic regimen until speciation and in vitro susceptibility testing documents equally effective alternatives. PMID- 8724956 TI - The microbiologic effect of gas filtration. AB - BACKGROUND: Air and expansile gases commonly are injected into the eye in both vitreoretinal surgery and anterior segment surgery. Although the halogenated gases are filtered routinely before injection, air often is not prepared in this manner. This study was designed to investigate the effect of filtration on the microbiology of gas mixtures. METHODS: Two separate studies were conducted, one examining medical grade perfluoropropane gas (C3F8), and the other evaluating air collected from an ophthalmic operating room or emergency room. Aliquots of air or gas were collected through a 0.22-micron filter before and after filtration. The filter discs were cultured for bacterial and fungal organisms. RESULTS: Of 514 samples collected, none grew either bacterial or fungal organisms. There was no difference with regard to recovered microbes between filtered and unfiltered air or C3F8, nor was there a difference between operating room air and emergency room air. Positive controls using aerosolized microorganisms yielded heavy growth. CONCLUSION: Based on results obtained in this study and available information on the concentration of airborne microbes, the microbiologic effect of gas filtration is of little significance for volumes routinely injected in ophthalmic surgery. PMID- 8724957 TI - Intraoperative echographic localization of iodine 125 episcleral radioactive plaques for posterior uveal melanoma. AB - PURPOSE: Plaque radiotherapy has been reported to have a higher relapse rate than charged-particle radiotherapy for posteriorly located uveal melanomas, which also are more technically difficult to localize accurately. The authors used intraoperative echography in patients with posterior uveal melanoma to determine the rate of inaccurate localization of iodine 125(125I) episcleral plaques using standard localization techniques. METHODS: The authors reviewed the records of 29 consecutive patients with medium-sized posterior uveal melanomas who underwent 125I episcleral plaque radiotherapy with intraoperative echographic verification of plaque placement. RESULTS: After careful plaque placement using standard localization techniques, 4 of 29 plaques (14%) did not cover at least one tumor margin. All four of these plaques were associated with posterior tumors with at least one margin posterior to the temporal arcades. Two (7%) additional juxtapapillary plaques were displaced away from the sclera by the optic nerve. In all six cases, it was possible to immediately reposition the plaque to achieve coverage of all tumor margins. CONCLUSIONS: Placement of 125I episcleral radioactive plaques for posteriorly located uveal melanomas using standard localization techniques occasionally results in suboptimal plaque positioning. Intraoperative echography can identify plaques that are localized poorly and allows immediate adjustment to achieve optimal plaque positioning. PMID- 8724958 TI - Retinoblastoma recurrence after iodine 125 plaque application. AB - PURPOSE: The authors report the clinicopathologic features of a recurrent retinoblastoma after radioactive iodine 125 (125l) plaque therapy. METHODS: The clinical and histopathologic findings of a 1-year-old boy with a recurrent retinoblastoma after 125l radioactive plaque application were reviewed. The patient's enucleated right eye was processed for routine histologic evaluation. Immunostains for glial fibrillary acid protein and S-100 protein were performed. RESULTS: Examination of the enucleated eye revealed a retinal tumor composed of two populations of cells. One population consisted of poorly differentiated cells, and the second population at the base of the tumor contained well differentiated cells with numerous fleurettes. Immunohistochemical stains were positive for glial fibrillary acid protein and S-100 protein in the well differentiated portion of the tumor. CONCLUSION: This case illustrates a recurrent retinoblastoma that may have arisen from radioresistant well differentiated cells after 125l plaque therapy. PMID- 8724960 TI - Clinicopathologic correlation of argon laser photocoagulation of retinal angiomas in a patient with von Hippel-Lindau disease followed for more than 20 years. AB - PURPOSE: The authors review the histopathologic findings in the eyes of a patient with multiple retinal angiomas and von Hippel-Lindau disease, who underwent treatment with argon laser photocoagulation with follow-up of more than 20 years. METHODS: The patient was studied ophthalmoscopically and by fluorescein angiography before and after argon laser photocoagulation of retinal angiomas. The eyes were obtained postmortem, and the central portion of the right eye, including the macula and optic nerve head, was sectioned serially for light microscopy. The pupil-optic nerve segment of the left eye was step-sectioned serially for light microscopy. RESULTS: Histopathologic study of the right eye disclosed mild cystoid macular edema and focal areas of exudation in the midperiphery possibly secondary to irradiation of the head. A 1.5 x 0.3-mm area of residual angioma was present in the nasal peripapillary retina. Superotemporally, four chorioretinal scars were present in one photocoagulated area. These scars were composed of dense fibrous tissue with vascularization and variable retinal pigment epithelium hyperplasia. Large, nonangiomatous vessels within each of the scars were continuous with other retinal vessels. Inferotemporally, two chorioretinal scars were present in one photocoagulated area. Histopathologically, these scars were similar to the superotemporal scars, except that no patent retinal vessels traversed the inferotemporal scars. Neovascularization of the retina was associated with one superotemporal and one inferotemporal scar. No residual angiomatous tissue was present in the supero- or inferotemporal areas. Histopathologic examination of the left eye disclosed extensive vitreous organization and periretinal fibrovascular proliferation, extensive gliosis of the retina, and a 4.5 x 2-mm schisis cavity filled with fibrinous exudate. Three angiomas with variable fibrosis were present in the left eye. CONCLUSION: Despite a poor clinical course in one eye treated with xenon arc photocoagulation, trans-scleral diathermy, and argon laser photocoagulation, a patient with von Hippel-Lindau disease and multiple retinal angiomas retained good vision in the other eye after successful treatment with argon laser photocoagulation with follow-up of more than 20 years. The amount of regression of angiomatous tissue after photocoagulation varied from lesion to lesion (complete in some; minimal in others). The authors conclude that argon laser photocoagulation of early lesions is effective in ablating smaller ( < or = 3 disc diameter) retinal angiomas. PMID- 8724959 TI - An experimental model of retinal pigment epithelial and neurosensory serous detachment. AB - PURPOSE: The authors document an animal model of retinal pigment epithelial (RPE) detachments. METHODS: N-ethylmaleimide (NEM) and sodium iodate (sulfhydryl alkylating agents) were injected into the vitreous of Dutch pigmented rabbits. Subretinal fluid samples were withdrawn 0.5 and 3 hours after NEM injections and analyzed for albumin content by gel electrophoresis. Selected eyes were examined histologically. RESULTS: Injections of 9.2 micrograms NEM caused wrinkling of the retina within 15 minutes, at which time fluorescein angiography revealed multifocal RPE detachments. At this stage, histologic sections showed moderate choroidal edema and RPE detachment. Elevated retinal detachments developed over the next 15 to 25 minutes. Histologic sections showed massive choroidal edema and RPE fragmentation. The albumin concentration of subretinal fluid rose from 60% of serum level at 30 minutes after NEM to 80% after 3 hours. Sodium iodate did not cause choroidal edema or RPE detachment. CONCLUSION: In rabbits, intravitreal NEM causes the rapid appearance of choroidal edema and RPE detachment, followed by serous retinal detachment. The initial effect probably is caused by an alteration of choroidal vascular permeability. The relationship of these effects to sulfhydryl alkylation is unclear because sodium iodate failed to produce RPE detachments. N-ethylmaleimide effects may model aspects of clinical RPE and serous retinal detachments. PMID- 8724961 TI - Diagnostic and therapeutic challenges. PMID- 8724962 TI - Detection of Epstein-Barr virus DNA by polymerase chain reaction in the vitreous from a patient with Vogt-Koyanagi-Harada syndrome. PMID- 8724963 TI - Simultaneous bilateral uveal effusions after cataract extractions. PMID- 8724964 TI - Capnocytophaga canimorsus endophthalmitis. PMID- 8724965 TI - Combined transscleral diode laser cyclophotocoagulation and transscleral retinal photocoagulation for refractory neovascular glaucoma. PMID- 8724966 TI - Five cases in which a depot steroid (hydrocortisone acetate and methylprednisolone acetate) was injected into the eye. PMID- 8724967 TI - Reticular pseudodrusen. PMID- 8724968 TI - Epiretinal membrane formation can occur in adult Coats' disease. PMID- 8724969 TI - Medial septal injection of naloxone elevates acetylcholine release in the hippocampus and induces behavioral seizures in rats. AB - The effects of injections of naloxone, a universal opioid receptor antagonist, into the medial septal nucleus on hippocampal acetylcholine (ACh) release and behavior were investigated in freely moving rats by means of the microdialysis method. The injection of naloxone (2, 10 and 20 micrograms) produced a marked increase in hippocampal ACh release in a dose-dependent manner. These effects of naloxone were reversed by the post-injection of [D-Ala2, N-Me-Phe4, Gly-ol] enkephalin (DAGO; 10 micrograms), an opioid mu receptor agonist. Furthermore, basal release of hippocampal ACh was significantly reduced by the injection of DAGO alone. It was also found that rats given an injection of naloxone showed an increase in motor activity and occasionally exhibited behavioral seizures. These effects of naloxone were also reversed by the post-injection of DAGO. The present results suggest that endogenous opioids ionically inhibit the activity of septo hippocampal cholinergic neurons via mediation of mu opioid receptors in the medial septal nucleus. They also suggest that endogenous opioids modulate the incidence of seizures, at least in part, through opioid mu receptors in the medial septal nucleus. PMID- 8724970 TI - 2D-PAGE analysis: adrenergically regulated pineal protein AIP 37/6 is a phosphorylated isoform of cytosolic malate dehydrogenase. AB - The adrenergic transmitter norepinephrine (NE) dramatically increases the prominence of only two out of the hundreds of [35S]methionine-labeled pineal proteins resolved by two-dimensional polyacrylamide gel electrophoresis (2D PAGE). One of these regulated proteins is AIP 37/6 (37 kDa, pI approximately 6). The labeling of this protein is increased approximately 100-fold by NE. In the study presented here the identity of AIP 37/6 was investigated. The results of microsequencing, immunochemical analysis of 2D-PAGE blots and size exclusion chromatography indicate that AIP 37/6 is an isoform of cytosolic malate dehydrogenase (cMDH; approximately 36.3 kDa; pI approximately 6.5). Associated studies indicate that this isoform is phosphorylated whereas the bulk of cMDH is not. Cotranslational phosphorylation of cMDH is discussed. PMID- 8724971 TI - Effect of lesions of forebrain circumventricular organs on c-fos expression in the central nervous system to plasma hypernatremia. AB - Experiments were carried out on conscious adult male Wistar rats to investigate the effect of selective ablation of the subfornical organ (SFO), and/or the anteroventral third ventricular (AV3V) region on the induction of Fos in central structures in response to plasma hypernatremia. Fos induction, detected immunohistochemically, was used as a marker for neuronal activation. Intravenous infusions of hypertonic saline resulted in dense Fos-like immunoreactivity in several forebrain (paraventricular nucleus of the hypothalamus (PVH), supraoptic nucleus (SON), median preoptic nucleus (MnPO), medial preoptic nucleus, organum vasculosum of the laminae terminalis and (SFO) and brainstem (nucleus of the solitary tract, ventrolateral medulla, and parabrachial nucleus) structures. Intravenous infusions of the hypertonic saline solution into animals with lesions of either the SFO, the AV3V or both resulted in a decreased number of Fos-like immunoreactive neurons in the MnPO, PVH and SON. In addition, the number of Fos labeled neurons in the SON after lesions of both the SFO and the AV3V was significantly greater than that observed in isotonic saline infused controls. Finally, lesions of the forebrain circumventricular structures did not alter the Fos labeling in brainstem structures as a result of the infusion of the hypertonic solution. These data suggest that changes in plasma osmolality and/or concentration of sodium alter the activity of SON and brainstem neurons in the absence of afferent inputs from the SFO and AV3V. PMID- 8724972 TI - Antagonists have a greater selectivity for muscarinic receptor subtypes in intact cerebellar granule cells than in membranes. AB - A comparison of muscarinic acetylcholine receptor (mAChR) antagonist binding properties was made between intact cerebellar granule cell cultures and membranes prepared from these cells. [3H]quinuclidinyl benzylate (QNB) binding displacement by four mAChR antagonists was measured and the selectivities for m2- or m3-mAChRs estimated by curve fitting. For each antagonist, the preparation of membranes caused a subtype selective decrease in receptor affinity, as compared to intact cell binding. The m2-selective antagonists had lower affinities in membranes for m2- but not for m3-mAChR, while the m3-selective antagonists had lower affinities for m3- but not for m2-mAChR. As a result, the m2-mAChR selectivity of AF-DX 116 and methoctramine in membranes was 66- and 1.7-fold less than in intact cells, and the m3-mAChR selectivity of 4-DAMP and pFHHSiD was 2.4- and 3.9-fold less in membranes than in intact cells. The m3-mAChR selectivity of 4-DAMP in intact cells was unaffected by cytoskeletal depolymerization with cytochalasins and colchicine. We suggest that the changes in selectivity seen with cell disruption may be due to a loss of cellular factors which regulate receptor properties. Antagonists binding to receptors on intact cells may cause subtype-specific changes in the interaction of the mAChR with these factors. These data suggest that mAChR antagonist binding selectivity needs to be re-examined in intact cell systems. PMID- 8724973 TI - Effects of unilateral or bilateral lesions within the anteroventral third ventricular region on c-fos expression induced by dehydration or angiotensin II in the supraoptic and paraventricular nuclei of the hypothalamus. AB - This paper reports the effects of AV3V lesions on the pattern of c-fos induced by 24 h dehydration. As expected, bilateral electrolytic lesions within the AV3V region (the ventral median preoptic nucleus) suppressed water intake following 24 h water deprivation. C-fos expression was also suppressed in the supraoptic (SON) and (less completely) in the paraventricular (PVN) nuclei, but not in the subfornical organ (SFO). Unilateral lesions of the AV3V region suppressed c-fos expression in the ipsilateral SON, but this selective ipsilateral effect was less in the PVN. The SFO was again unaffected. Unilateral lesions also suppressed c fos expression in the ipsilateral SON and PVN (to a lesser degree) following intraventricular infusions of angiotensin II (250 pmol). These results suggest that the cellular response of supraoptic neurons to osmotic stimuli require inputs from the AV3V region, but that this is less absolute for the PVN; that the projection from the ventral AV3V area to the SON is ipsilateral, but that to the PVN may be less lateralised. Activation of the SFO by dehydration is not dependent upon the integrity of the ventral AV3V region. These results are closely comparable to the effects of similar lesions on c-fos expression following intraventricular infusions of angiotensin II, and suggest that the effect of dehydration on forebrain c-fos expression may be related to the central actions of angiotensin II. PMID- 8724974 TI - Effects of haloperidol on the activity and membrane physiology of substantia nigra dopamine neurons recorded in vitro. AB - A variety of experimental evidence suggests that one function of dendritically released dopamine is the feedback modulation of dopamine neuron firing rate via stimulation of the somatodendritic autoreceptors located on these cells. Under these conditions, blockade of these receptors should result in an alteration in the firing rate of dopamine neurons. In order to test this prediction, we have examined whether haloperidol alters the electrophysiological activity of dopamine neurons recorded from nigral slices maintained in vitro. This preparation permits examination of the effects of haloperidol when the substantia nigra is isolated from long-loop afferent cell populations, and also facilitates the performance of intracellular recordings to allow the assessment of alterations in membrane properties that underlie any changes in firing rate. Addition of haloperidol to the media bathing nigral slices caused increases in the spontaneous firing rate of some dopamine neurons. It also caused depolarization of the membrane and increases in input resistance in a subset of dopamine neurons. However, this drug had no consistent effects on the delayed repolarization or the anomalous rectification that are characteristic of activity in these cells. Morphological assessment of dopamine neurons stained in the coronal sections used in these studies confirmed that the dendrites of dopamine neurons were largely intact in these slices. These results demonstrate that blockade of the somatodendritic autoreceptors located on dopamine neurons does cause alterations in the electrophysiological activity of these cells, substantiating the role of nigral dopamine release in the modulation of dopamine neuron activity. PMID- 8724975 TI - SC1, a SPARC-related glycoprotein, exhibits features of an ECM component in the developing and adult brain. AB - Although extracellular matrix (ECM) components have been shown to play important roles in the development of the CNS, expression generally decreases in the adult brain. This study examines the expression of the SPARC-related glycoprotein SC1 in the rat brain during postnatal development and in the adult. In situ hybridization analysis indicates that expression of SC1 mRNA increases in a caudal to rostral manner as postnatal neural development proceeds and is found at near maximal levels in the adult brain. SC1 mRNA is expressed in glial-enriched areas of the brain at postnatal day 1 (P1) and P5. Between P10 and P20, SC1 mRNA increases in neuron-enriched regions of the hippocampus, dentate gyrus, and cerebral cortex. Immunohistochemistry in the adult shows that SC1 protein is localized to neurons in these regions and to scattered glial cells. Subcellular fractionation demonstrates that the SC1 116/120 kDa doublet is associated with synaptosomes. SC1 is present in the aqueous phase following extraction of membranes with TX-114, suggesting that it is not a transmembrane protein, a property consistent with other adult brain ECM components. Furthermore in cerebellar granule cells grown in culture, high levels of the 120 kDa component are secreted into the media. These results are consistent with the hypothesis that SC1 is an ECM glycoprotein expressed in both the developing and adult brain. PMID- 8724976 TI - Expression of inducible cyclooxygenase mRNA in the mouse brain after systemic administration of bacterial lipopolysaccharide. AB - Cyclooxygenase (COX) is an enzyme involved in the biosynthesis of prostaglandins and is one of the principle targets of non-steroidal anti-inflammatory drugs. Two isoforms of this enzyme are known to exist in the brain; one of these (type 1 COX or COX1) is constitutively expressed, whereas the other form of the enzyme, which is inducible, has been called type 2 COX (COX2). We have used systemic administration of bacterial lipopolysaccharide (LPS) as a model of the acute phase response to study the expression of COX2 in the murine CNS. We observed COX2 expression in neurons of several regions of the normal murine telencephalon. Robust expression of COX2 mRNA was induced in perivascular cells between 45 min and 6 h after LPS injection. The role of prostaglandins produced by these perivascular cells in the cerebral components of the acute phase response remains to be elucidated. PMID- 8724977 TI - Modulatory action of dopamine on acetylcholine-responsive striatal and accumbal neurons in awake, unrestrained rats. AB - In ambulant rats, iontophoresis of low concentrations of dopamine (DA) enhances the response of neurons in striatum and nucleus accumbens to iontophoretic glutamate. In an extension of this line of investigation, we tested the effects of acetylcholine (ACh), a presumed modulator of neuronal function in these same brain regions, and assessed possible DA-ACh interactions. Data were obtained from spontaneously active neurons known to respond to ACh (5-30 nA) when the animals rested quietly with no overt movement. ACh iontophoresis either excited or inhibited striatal and accumbal activity but excitatory effects predominated in both areas. With multiple applications of ACh, especially at the lowest currents tested, either response often was interspersed with instances of no change in firing rate. Responsiveness to ACh also diminished during periods of spontaneous movement when basal firing showed phasic increases in activity. In fact, neurons with the highest rates of basal activity showed the smallest magnitude response to ACh. Prolonged applications (120-180 s) of DA attenuated basal firing as well as the iontophoretic effects of ACh both during the DA application itself and for up to 1 min after DA ejection offset. The result of these inhibitory effects was no net change in the relative magnitude of the ACh response. Thus, although ACh can modulate striatal and accumbal neuronal activity, DA does not regulate this effect in the same way that it regulates the neuronal responsiveness to glutamate. PMID- 8724978 TI - Neuroanatomical characterization of Fos induction in rat behavioral models of anxiety. AB - Immunohistochemical staining for Fos-like immunoreactivity (Fos-LI) was used to map functional activation in discrete brain regions of rats processed in three empirical models of anxiety: foot shock avoidance responding in a shuttle box, the elevated plus maze, and an air puff-induced ultrasonic vocalization test. The avoidance test and elevated plus maze induced prominent Fos-LI in select brain regions, including the medial prefrontal, cingulate, and ventrolateral orbital cortices, taenia tecta, nucleus accumbens, paraventricular nucleus of the hypothalamus, medial nucleus of the amygdala and lateral septum. Air puff stimuli that produced ultrasonic vocalizations induced Fos-LI to a more limited extent compared to the plus maze and avoidance test, with only the medial prefrontal cortex, medial nucleus of the amygdala, and lateral septum being significantly affected by air-puff. Even though the sensory stimuli and environmental conditions associated with the three anxiety models were markedly different, specific common forebrain regions were affected, i.e. the medial prefrontal cortex, medial amygdala, and lateral septum. It is hypothesized that these regions are components of a circuit in the rat brain related to anxiety or distress. To determine the potential relationship between generalized arousal and the observed induction of Fos-LI in the anxiety models, rats were tested in a non aversive situation involving marked behavioral activation. Accordingly, after vigorous bar pressing behavior for reinforcement with sweetened condensed milk, induction of Fos-LI was minimal and comparable to that in unhandled control rats. These latter data indicate that the distinctive neuroanatomical patterns of Fos LI observed in the paradigms related to anxiety were not simply due to generalized behavioral activation. In summary, select common brain regions were identified that express Fos-LI in empirical models of anxiety. These data provide a functional framework to explore neuroanatomical sites of action of psychotherapeutic drugs that influence behavioral responses in these tasks. PMID- 8724979 TI - Effect of 6R-L-erythro-5,6,7,8-tetrahydrobiopterin and infusion of L-tyrosine on the in vivo L-[beta-11C] DOPA disposition in the monkey brain. AB - The effect of 6R-L-erythro-5,6,7,8-tetrahydrobiopterin (6R-BH4) and L-tyrosine infusion on [11C]dopamine synthesis was analyzed in the striatum of Rhesus using positron emission tomography (PET). The rate for decarboxylation from L-[beta 11C]DOPA to [11C]dopamine was calculated using a graphical method with cerebellum as a reference region. Although the peripheral administration of 6R-BH4 at low dose (2 mg/kg) did not provide a significant increase in the rate of dopamine biosynthesis, a high dose of 6R-BH4 (20 mg/kg) induced an elevation of the rate. This 6R-BH4-induced elevation of the dopamine synthesis rate was further dose dependently enhanced by the continuous infusion of L-tyrosine (0.2 and 1.0 mumol/min/kg). L-Tyrosine infusion with a rate of 1.0 mumol/min/kg caused an enhancement of the rate even during low dose administration of 6R-BH4 (2 mg/kg). L-Tyrosine infusion alone did not induce any elevation of the dopamine biosynthesis rate. The analysis of plasma indicated that the metabolic ratios of L-[beta-11C]DOPA to each metabolite were not affected by 6R-BH4 and/or L-tyrosine infusion. The results suggest that the low dose loading of tyrosine facilitates the activity of 6R-BH4 on the presynaptic dopamine biosynthesis, and also that the combined effects can be monitored by PET using L-[beta-11C]DOPA as a biochemical probe. PMID- 8724980 TI - Differential and persistent expression patterns of CNS gene transfer by an adeno associated virus (AAV) vector. AB - Safe, long-term gene expression is a primary criteria for effective gene therapy in the brain, so studies were initiated to evaluate adeno-associated virus (AAV) vector transfer of a reporter gene into specific sites of the rat brain. In the 4 day old rat, site infusions of AAV-CMV-lacZ (1 microliter; 5 x 10(4) particles) produced neuronal beta-galactosidase gene expression 3 weeks later in the hippocampus and inferior colliculus, but not in the cerebral cortex. Seven days after infusion of AAV-CMV-lacZ viral vectors (1 microliter) in the adult rat, beta-galactosidase gene expression was found in the olfactory tubercle, caudate, hippocampus, piriform cortex and inferior colliculus. primarily in multipolar neurons close to the infusion site. Three months after vector microinfusion, similar levels of gene expression remained in the olfactory tubercle and the inferior colliculus, with some reduction found in the caudate, but substantial reductions in beta-galactosidase gene expression occurred in the hippocampus and piriform cortex. In no case were obvious signs of toxicity noted. Therefore, AAV vectors can transfer foreign genes into the adult and neonatal CNS, but the pattern and longevity of gene expression depends upon the area of brain being studied. PMID- 8724981 TI - Effects of fluoxetine and olfactory bulbectomy on mouse circadian activity rhythms. AB - Olfactory bulbectomy (OBX) in SWR outbred male mice lengthened the free-running period and delayed the phase of a circadian rhythm for wheel-running activity. OBX also increased mean levels of activity. Two weeks of daily intraperitoneal injections of Fluoxetine (8 mg/kg), a serotonin re-uptake inhibitor, reversed the effects of bulbectomy on the mean level of activity and significantly shortened the free-running period of the activity rhythm. The phase of the activity rhythm was not significantly affected by the Fluoxetine treatment. These results are consistent with a hyposerotonergic mediation of the effects of OBX on circadian period and activity level. PMID- 8724982 TI - Effect of nucleus accumbens dopamine depletion on motivational aspects involved in initiation of cocaine and heroin self-administration in rats. AB - The involvement of mesolimbic dopamine (DA) systems in motivational aspects of drug-taking behavior during initiation of drug self-administration was investigated using a recently developed behavioral paradigm. In separate experiments animals were allowed to self-administer cocaine or heroin (0.16 and 0.32 mg . kg-1 per inf) during 5 consecutive daily 3-h sessions. During a 15-min period preceding the last four self-administration sessions lever-press behavior was measured in absence of the drug as an index of the motivational aspects involved in drug-taking behavior. The effect of 6-hydroxydopamine (6-OHDA) lesion of the nucleus accumbens (NAC) on lever-press behavior before and during self administration was measured. Destruction of DA terminals in the NAC did not affect initiation of heroin self-administration nor the lever-press behavior during the period preceding the self-administration sessions. In cocaine animals 6-OHDA lesion of the NAC decreased the total intake of cocaine during the self administration sessions and impaired discriminative lever-responding for the drug, both during cocaine self-administration, and during preceding periods when no cocaine was available. It is concluded that DAergic systems in the NAC might be involved in the reinforcement and/or motivational processes underlying cocaine self-administration. The present findings, however, do not support the notion of a critical role of NAC DA in the motivational aspects of drug-taking behavior in general. PMID- 8724983 TI - Regulation of tyrosine hydroxylase and tetrahydrobiopterin biosynthetic enzymes in PC12 cells by NGF, EGF and IFN-gamma. AB - The regulation of catecholamine and tetrahydrobiopterin synthesis was investigated in cultured rat pheochromocytoma PC12 cells following treatments with nerve growth factor (NGF), epidermal growth factor (EGF) and interferon gamma (IFN-gamma). NGF and EGF, but not IFN-gamma, caused an increase after 24 h in the levels of BH4 and catecholamines, and the activities of tyrosine hydroxylase and GTP cyclohydrolase, the rate-limiting enzymes in catecholamine and BH4 synthesis, respectively. Actinomycin D, a transcriptional inhibitor, blocked treatment-induced elevations in tyrosine hydroxylase and GTP cyclohydrolase activities. NGF, EGF or IFN-gamma did not affect the activity of sepiapterin reductase, the final enzyme in BH4 biosynthesis. Rp-cAMP, an inhibitor of cAMP-mediated responses, blocked the induction of tyrosine hydroxylase by NGF or EGF; inhibition of protein kinase C partially blocked the EGF effect, but not the NGF effect, NGF also induced GTP cyclohydrolase in a cAMP dependent manner, while the EGF effect was not blocked by Rp-cAMP or protein kinase C inhibitors. Sphingosine induced GTP cyclohydrolase in a protein kinase C independent manner without affecting tyrosine hydroxylase activity. Our results suggest that both tyrosine hydroxylase and GTP cyclohydrolase are induced in a coordinate and transcription-dependent manner by NGF and EGF, while conditions exist where the induction of tyrosine hydroxylase and GTP cyclohydrolase is not coordinately regulated. PMID- 8724984 TI - Electrophysiological actions of hemoglobin on rat hippocampal CA1 pyramidal neurons. AB - Hemoglobin, the oxygen-carrying component of red blood cells, can be released from erythrocytes in hemorrhagic stroke and intracranial bleeding associated with head injuries. Therefore, neurons may be exposed to this agent. In addition, hemoglobin can chelate nitric oxide (NO) and has been used in studying the role of NO in synaptic plasticity and excitotoxicity. However, the electrophysiological actions of hemoglobin on central neurons are not well characterized. In the present investigation, the electrophysiological actions of hemoglobin on CA1 pyramidal neurons in rat hippocampal slices were studied with conventional intracellular pointed microelectrode- as well as perforated patch recordings. Superfusion of rat hippocampal slices with hemoglobin (0.05 or 0.1 mM for 10-15 min) induced a depolarization of CA1 neurons and suppressed the stratum radiatum stimulation-induced excitatory postsynaptic potentials (EPSPs) and inhibitory postsynaptic potentials (IPSPs). The hemoglobin-induced depolarization as well as the suppression of the synaptic transients were present in slices pretreated with 0.1 or 0.5 mM of N omega-nitro-L-arginine, a nitric oxide synthase inhibitor, suggesting that hemoglobin has electrophysiological actions on hippocampal CA1 neurons that are independent of its NO scavenging property. PMID- 8724985 TI - Interleukin-6 released in human cerebrospinal fluid following traumatic brain injury may trigger nerve growth factor production in astrocytes. AB - Cytokines are involved in nerve regeneration by modulating the synthesis of neurotrophic factors. The role played by interleukin-6 (IL-6) in promoting nerve growth factor (NGF) after brain injury was investigated by monitoring the release of IL-6 and NGF in ventricular cerebrospinal fluid (CSF) of 22 patients with severe traumatic brain injuries. IL-6 was found in the CSF of all individuals and remained elevated for the whole study period. NGF appeared in the CSF if IL-6 levels reached high concentrations and was often detected simultaneously with or following an IL-6 peak. The amounts of NGF correlated with the severity of the injury, as indicated by the clinical outcome of the patients. The functional relationship of IL-6 and NGF was investigated utilizing cultured mouse astrocytes. The CSF of 8 patients containing IL-6 induced NGF production in astrocytes, whereas control CSF without IL-6 had no effect. The induction of NGF was inhibited up to 100% by adding anti-IL-6 antibodies. These results were corroborated when astrocytes were exposed to recombinant IL-6 at different concentrations resulting in NGF production. Thus, the production of IL-6 within the injured brain may likely contribute to the release of neurotrophic factors by astrocytes. PMID- 8724986 TI - The role of limbic vasopressin and oxytocin in social recognition. AB - Young male rats were exposed two times for 5 min, to older male rats with an interval of 30 min in the anti-vasopressin serum experiments and with an interval of 120 min in the anti-oxytocin serum experiments. The time spent by the older rats with social investigation of the younger animal was scored during the two encounters. In placebo-treated animals the time spent on social investigation of the younger animal during the second encounter at 30 min is significantly shorter than during the first one (social recognition). However, intracerebroventricular or local application of anti-vasopressin serum in the dorsal or ventral hippocampus or in the dorsal septal area, but not in the n. olfactorius, results in similar periods of time spent for social investigation during the two encounters. Thus, endogenous vasopressin in the dorsal and ventral hippocampus and in the dorsal septal region plays a physiological role in social recognition/memory. In placebo-treated rats the time spent on social investigation of the younger animal during the second encounter at 120 min is similar to that during the first encounter. However, local administration of anti oxytocin serum in the ventral hippocampus, but not in the dorsal hippocampus, nor in the n. olfactorius or the septal area, results in shorter investigation times during the second encounter. Thus, taken together the presence or local release of vasopressin and oxytocin in the ventral hippocampus and that of vasopressin (but not oxytocin) in the dorsal hippocampus and dorsal septal area are of physiological importance for social recognition. PMID- 8724987 TI - Tachykinins cause inward current through NK1 receptors in bullfrog sensory neurons. AB - The effects of tachykinins on primary afferent neurons of bullfrog dorsal root ganglia (DRG) were examined by using whole-cell patch-clamp methods. Neurokinin A (NKA) caused inward current (INKA) in a concentration-dependent manner. Concentration-response curve showed that the EC50 for NKA was 6 nM. The INKA showed strong tachyphylaxis, when NKA was continuously applied for more than 1 min. Substance P (SP) also produced inward current with potency similar to that of NKA. Neurokinin B (NKB) was less effective in producing the inward current. The order of agonist potency was NKA = SP >> NKB. Spantide ([D-Arg1, D-Trp7.9, Leu11]SP), a non-selective peptide antagonist at tachykinin receptors, reduced the tachykinin-induced current. CP-99,994, a selective non-peptide antagonist for neurokinin-1 (NK1) receptor, inhibited the inward currents produced by NKA and SP. The INKA was associated with decrease in K+ conductance. NKA suppressed both a voltage-dependent K+ current, the M-current (IM), and a voltage-independent background K+ current, IK(B). Intracellular dialysis with GTP gamma S (100 nM) or GDP beta S (100 microM) depressed the INKA. Pre-treatment of DRG neurons with pertussis toxin (PTX) did not prevent the INKA. Depletion of intracellular ATP depressed the INKA. These results suggest that the tachykinin-induced inward current is mediated through the NK1 receptor which mainly couples to PTX insensitive G-protein in bullfrog primary afferent neurons. PMID- 8724988 TI - Distribution of NADPH-diaphorase in the central nervous system of an infrared sensitive snake, Trimeresurus flavoviridis. AB - The distribution of NADPH-diaphorase (NADPH-d) activity was studied in the central nervous system of an infrared sensitive snake. An inhibitor of nitric oxide synthase, dichloroindophenol (DPIP), was used to distinguish the characteristics of NADPH-d activity. Intensely and weakly NADPH-d-stained neurons and fibers were found in discrete regions throughout the snake brain and cervical spinal cord, such as the olfactory bulb, subcommissural organ, stratum griseum periventriculare, locus coeruleus, dorsal root, dorsal horn, and area X. It was particularly noticed that the trigeminal descending nuclei and reticular formation of the medulla oblongata contained many positive neurons and fibers, but the lateral descending nucleus and nucleus reticularis caloris (infrared sensory nuclei) certainly did not. The positive neurons and fibers were also observed in supraspinal sensory ganglia. DPIP inhibited NADPH-d activity in all regions except for the olfactory/vomeronasal nerve and glomeruli. The results prove for the first time the presence of NADPH-d activity in the ophidian brain and suggest that nitric oxide may be involved in many neural functions, but not in infrared sensory processing. PMID- 8724989 TI - NGF, BDNF and NT-5, but not NT-3 protect against MPP+ toxicity and oxidative stress in neonatal animals. AB - A growing body of evidence suggests that neurotrophic factors can protect neurons against neuronal death. In the present study we examined whether systemic administration of members of the neurotrophin family, nerve growth factor (NGF), brain derived neurotrophic factor (BDNF), neurotrophin 3 (NT-3) and neurotrophin 5 (NT-5) and basic fibroblast growth factor (bFGF) could protect against 1-methyl 4-phenylpyridinium (MPP+) induced striatal damage in neonatal rats. Systemic administration of NGF, BDNF and NT-5 produced significant neuroprotective effects, whereas NT-3 was ineffective. Systemic administration of bFGF had significant neuroprotective effects as assessed by T2-weighted magnetic resonance imaging and measurements of n-acetylaspartate and lactate using chemical shift magnetic resonance imaging. Systemic administration of NGF, BDNF and bFGF, but not NT-3 attenuated MPP+ induced increases in hydroxyl radical generation as assessed by the conversion of salicylate to 2,3- or 2,5-dihydroxybenzoic acid (DHBA). These results show that systemic administration of several neurotrophins and bFGF can attenuate neuronal damage induced by chemical hypoxia in vivo by a mechanism which may involve attenuation of oxidative stress. PMID- 8724990 TI - Acceleration deficit in patients with cerebellar lesions. A study of kinematic and EMG-parameters in fast wrist movements. AB - Slowness of goal-directed movements is a frequent symptom following cerebellar lesions. So far it has not been demonstrated whether this slowness represents compensation for impaired braking which is a feature of cerebellar dysfunction with the consequence of hypermetria, or whether it is an independent part of cerebellar movement disorder. To resolve this question we tested 18 cerebellar patients in a paradigm where they not only had to perform fast goal-directed wrist flexion movements (amplitudes 5 degrees and 30 degrees) but also wrist flexion movements as fast and large as possible without particular target. In normals antagonist activity is minimal in large movements without target. Although subjects were clinically only mildly affected they regularly showed a 'slowness of movement' resulting from a reduction of peak acceleration. This in turn was due to the reduced generation of agonist activity. Peak velocity was not significantly decreased because the acceleration phase was adequately prolonged. Since these changes were most pronounced in the 'fast' movements without target the compensation hypothesis should be discarded. The reduction of acceleration must at least partially be due to a genuine cerebellar deficiency in the fast generation of agonist activity. PMID- 8724991 TI - Selective induction of interleukin-6 in mouse microglia by granulocyte-macrophage colony-stimulating factor. AB - Astrocytes produce granulocyte/macrophage colony-stimulating factor (GM-CSF) and support the survival and proliferation of microglia. To study the functions of GM CSF in the central nervous system (CNS), we examined the effects of GM-CSF on cytokine production by glial cells. GM-CSF induced interleukin-6 (IL-6) production by microglia, but not by astrocytes, in a dose-dependent manner as assessed by bioassay and the detection of IL-6 mRNA by reverse transcriptase polymerase chain reaction (RT-PCR) analysis. GM-CSF did not induce tumor necrosis factor (TNF) alpha or IL-1 in microglia and astrocytes, whereas lipopolysaccharide induced all these cytokines. The induction of IL-6 by GM-CSF in microglia was completely inhibited by antibodies to GM-CSF. Neither IL-3 nor macrophage-CSF (M-CSF) induced IL-6 production in microglia. Given that IL-1 and TNF alpha, monokines derived from microglia, induce IL-6 production in astrocytes, but not in microglia, results indicate that astrocytes and microglia may mutually regulate IL-6 production by different cytokines. PMID- 8724992 TI - Temporal profile of nerve growth factor-like immunoreactivity after transient focal cerebral ischemia in rats. AB - We studied the temporal profile of nerve growth factor-like immunoreactivity (NGF LI) in the rat brains following 30 min of middle cerebral artery occlusion. The rats were decapitated at 4 h, 1, 3, 7, and 14 days of recirculation. Brain sections at the level of striatum were immunostained against NGF as well as a stress protein, HSP70. Also, double immunostaining of NGF and glial fibrillary acidic protein was performed. In the sham-control rats, NGF-LI was normally present in the cortical and striatal neurons. However, at 4 h of recirculation, there was a significant decrease of NGF-LI in the ischemic cortex and striatum. From 1 day, NGF-LI was absent completely in the ischemic striatum. However, in the ischemic cortex, NGF-LI decreased to the lowest level at 1 day, but it recovered gradually from 3 days and increased significantly to above sham-control level at 7 days. At 14 days of recirculation, NGF-LI returned to a near sham control level. In the non-ischemic cortex, NGF-LI increased gradually from 4 h with a peak at 7 days, and returned to the sham-control level at 14 days of recirculation. A HSP70 was induced in the ischemic cortex at 1 and 3 days, when there was a significant reduction of NGF-LI. The number of reactive astrocytes increased gradually and NGF-LI in the reactive astrocytes became gradually intense after ischemia. The present finding showing that NGF-LI can be recovered in the stressed cortical neurons suggests a possible involvement of NGF in the process of neuronal survival after focal cerebral ischemia. The expression of NGF in reactive astrocytes indicates that astrocyte may also play a role in supporting neuronal survival after ischemia. PMID- 8724993 TI - Heme-oxygenase-1 induction in glia throughout rat brain following experimental subarachnoid hemorrhage. AB - The heme released following subarachnoid hemorrhage is metabolized by heme oxygenase (HO) to biliverdin and carbon monoxide (CO) with the release of iron. The HO reaction is important since heme may contribute to vasospasm and increase oxidative stress in cells. HO is comprised of at least two isozymes, HO-2 and HO 1. HO-1, also known as heat shock protein HSP32, is inducible by many factors including heme and heat shock. HO-2 does not respond to these stresses. To begin to examine HO activity following subarachnoid hemorrhage (SAH), the expression of HO-1 and HO-2 was investigated after experimental SAH in adult rats. Immunocytochemistry for HO-1, HO-2 and HSP70 proteins was performed at 1, 2, 3 and 4 days after injections of lysed blood, whole blood, oxyhemoglobin and saline into the cisterna magna. A large increase in HO-1 immunoreactivity was seen in cells throughout brain following injections of lysed blood, whole blood, and oxyhemoglobin but not saline. Lysed blood, whole blood and oxyhemoglobin induced HO-1 in all of the cortex, hippocampus, striatum, thalamus, forebrain white matter and in cerebellar cortex. HO-1 immunoreactivity was greatest in those regions adjacent to the basal subarachnoid cisterns where blood and oxyhemoglobin concentrations were likely highest. Double immunofluorescence studies showed the HO-1 positive cells to be predominately microglia, though HO-1 was induced in some astrocytes. HO-1 expression resolved by 48 h. HO-2 immunoreactivity was abundant but did not change following injections of blood. A generalized induction of HSP70 heat shock protein was not observed following injections of lysed blood, whole blood, oxyhemoglobin, or saline. These results suggest that HO 1 is induced in microglia throughout rat brain as a general, parenchymal response to the presence of oxyhemoglobin in the subarachnoid space and not as a stress response. This microglial HO-1 response could be protective against the lipid peroxidation and vasospasm induced by hemoglobin, by increasing heme clearance and iron sequestration, and enhancing the production of the antioxidant bilirubin. PMID- 8724994 TI - Central projections and entries of capsaicin-sensitive muscle afferents. AB - The entry pathway and central distribution of A delta and C muscle afferents within the central nervous system (CNS) were investigated by combining electron microscopy and electrophysiological analysis after intramuscular injection of capsaicin. The drug was injected into the rat lateral gastrocnemius (LG) and extraocular (EO) muscles. The compound action potentials of LG nerve and the evoked field potentials recorded in semilunar ganglion showed an immediate and permanent reduction in A delta and C components. The morphological data revealed degenerating unmyelinated axons and terminals in the inner sublamina II and in the border of laminae I-II of the dorsal horn at L4-L5 and C1-C2 (subnucleus caudalis trigemini) spinal cord segments. Most degenerating terminals were the central bouton (C) of type I and II synaptic glomeruli. Furthermore, degenerating peripheral axonal endings (V2) presynaptic to normal C were found. Since V2 were previously found degenerated after cutting the oculomotor nerve (ON) or L4 ventral root, we conclude that some A delta and C afferents from LG and EO muscles entering the CNS by ON or ventral roots make axoaxonic synapses on other primary afferents to promote an afferent control of sensory input. PMID- 8724995 TI - Arginine vasopressin reduces the blood-brain transfer of L-tyrosine and L-valine: further evidence of the effect of the peptide on the L-system transporter at the blood-brain barrier. AB - Arginine vasopressin (AVP) coinjected into the carotid artery in physiological concentrations (0.1 nmol/l), with either L-[3H]tyrosine or L-[3H]valine, induced changes in the kinetic parameters of the blood-to-brain transfer of both large neutral amino acids (LNAA) without alterations in brain haemodynamics. The half saturation constant (Km), the maximum velocity of transport (V(max)) and Kd, the nonsaturable transport constant, were estimated in 9 brain regions of male Wistar rats anaesthetized with ether. Apart from Kd, significant changes in Km and V(max) were observed in all brain regions investigated. On average Km decreased from 0.17 to 0.048 mmol/l for tyrosine, and from 0.61 to 0.059 mmol/l for valine, whereas V(max) declined from 22 to 9.9 nmol/min/g for tyrosine, and from 29 to 3.2 nmol/min/g for valine, respectively. The results provide further evidence that vasopressin-receptor interactions at the blood-brain barrier (BBB) induce changes in the properties of the common transporter, the L-system, which eventually result in a suppression of the blood-to-brain transfer of LNAA. Data analysis of the 5 LNAA tested so far reveals a significant negative correlation (R = 0.98, P < 0.05) between the respective substrate affinity for the transporter and the corresponding magnitude of transport reduction induced by circulating AVP. Calculations of the unidirectional influx (J) of the LNAA indicate that AVP (1) reduces J by approximately one-third for every LNAA, but (2) does not change the relative contribution for each single LNAA to the total influx across the BBB. PMID- 8724996 TI - Lipoprotein that selectively inhibits taste nerve responses to bitter substances. AB - The development of a specific inhibitor for bitter taste has been widely required in the fields of taste physiology and pharmaceutical sciences, but no inhibitor has been available. We found that lipoproteins, PA-LG composed of phosphatidic acid (PA) and beta-lactoglobulin (LG) and PA-LA composed of PA and alpha lactalbumin (LA) reversibly suppressed the responses of the frog glossopharyngeal nerve to the bitter substances. The frog tongue was treated with PA-LG solution for 10 min and then stimulated by a stimulus dissolved in water. The responses to the bitter substances such as quinine hydrochloride, papaverine hydrochloride, caffeine and L-leucine were completely suppressed by PA-LG, while those to the salt type bitter substances such as CsCl, MgCl2 and tetraethylammonium chloride were not suppressed. The responses to NaCl, galactose, acetic acid and L-alanine were unchanged or only slightly increased. The results suggested that binding of PA-LG to the hydrophobic region of the receptor membranes leads to suppression of the responses to the bitter substances. It was pointed out that PA-LG is useful not only for elucidating the receptor mechanisms of bitter substances, but also can be safely used to mask the bitter taste of foods and drugs, since PA, LG and LA are prepared from foods (soybean and milk). PMID- 8724997 TI - FR discrimination training reverses 6-hydroxydopamine-induced striatal dopamine depletion in a rat model of Lesch-Nyhan syndrome. AB - Five-day-old rats received 6-hydroxydopamine (6-HD; 100 micrograms base) or vehicle intracisternally. Striatal and cortical dopamine (DA) and metabolite levels were then determined when animals were three or 8.5 months of age and the latter rats had been weight-reduced for 5.5 months. In the latter animals these determinations were made 1 month following 4.5 months of home-cage confinement (untrained animals) or of food-maintained fixed-ratio (FR) discrimination training involving either a single discrimination (performance animals) or incrementally more difficult discriminations. Striatal DA levels in 3-month-old and 8.5-month-old (untrained) 6-HD-treated rats were, respectively, only 3% and 11% of those in untrained vehicle-treated animals (controls). Despite such large depletions, striatal DA levels in 6-HD-treated performance rats were 3-fold higher than those in untrained age-matched 6-HD-treated rats (i.e., were 32% of values in controls) while those in incrementally trained 6-HD-treated animals were even higher (i.e., were 60% of control values). Related changes occurred in levels of most metabolites. However, in incrementally trained rats, striatal 3 methoxytyramine concentrations were 154% of control values. Cortical DA and metabolite levels were little affected by 6-HD treatment. The present results confirm and extend our earlier observations suggesting that reversal of 'irreversible' neonatal 6-HD-induced striatal dopamine and metabolic depletion can be accomplished by environmental (training) manipulations in adult rats. PMID- 8724998 TI - Brainstem stimulation during sleep evokes abnormal rhythmic activity in thalamic neurons in feline penicillin epilepsy. AB - Some periods in the sleep-waking cycle are more seizure prone than others. In absence epilepsy, transition periods between nonrapid-eye-movement (nonREM) sleep and waking or REM sleep can be more seizure prone that stable states. One feature of transition periods that is hypothesized to promote seizure activity is the presence of coincident activity in ascending brainstem reticular formation (RF) arousal systems with synchronized thalamo-cortical activity. To evaluate this hypothesis we examined the state-dependent effects of low intensity RF stimulation on thalamic single unit activity in control conditions and following systemic penicillin-G administration to adult cats. In control conditions, RF stimulation during waking and REM sleep typically evoked a short-latency action potential in thalamic neurons. The same stimulation during nonREM sleep commonly evoked a high frequency burst of action potentials followed by a period of suppressed discharge. In 16/26 neurons, a second rebound burst of action potentials followed the period of discharge suppression. The average interval between the initial and rebound bursts was 75.1 +/- 6.0 ms, which was similar to the interburst interval recorded in these same cells during spontaneous EEG spindles. Following administration of penicillin-G, RF stimulation during nonREM sleep evoked high frequency burst firing, followed by 1-2 rebound bursts in 21/22 thalamic neurons. The average evoked interburst interval was 152.5 +/- 7.3 ms, a value comparable to the interburst interval displayed by these same cells during spontaneous spike-wave seizure activity (157.8 +/- 8.7 ms). RF-evoked rhythmic discharges were dependent upon the presence of thalamocortical synchronization, as responses evoked during waking and REM sleep in penicillin treated cats were similar to those observed in control conditions. PMID- 8724999 TI - Norepinephrine depletion impairs neuroendocrine responses to fear but not novel environmental stimuli in the rat. AB - Emotional stimuli suppress vasopressin secretion and potentiate oxytocin and prolactin secretion by the pituitary in the rat. We studied effects of central norepinephrine depletion on these hormonal responses to novel environmental or fear stimuli. Male Wistar rats were injected intracerebroventricularly with 5 amino-2,4-dihydroxy-alpha- methylphenylethylamine, a selective neurotoxin to noradrenergic fibers. The neurotoxin treatment reduced the hypothalamic content of norepinephrine by 71% but did not significantly affect the dopamine content. Novel environmental stimuli suppressed vasopressin secretion and augmented secretion of oxytocin and prolactin in the vehicle-injected rats. The neurotoxin did not block the neuroendocrine responses. Intermittently applied electric footshocks also induced the similar neuroendocrine responses in the vehicle injected rats. The neurotoxin significantly reduced the neuroendocrine responses. The drug, however, did not significantly alter vasopressin release after continuously applied footshocks. Environmental stimuli previously paired with footshocks (conditioned fear stimuli) suppressed vasopressin secretion and augmented secretion of oxytocin and prolactin in the vehicle-injected animals. Motor activity was suppressed during the conditioned fear stimuli. The neurotoxin impaired the neuroendocrine and behavioral responses whether the drug was injected before or after the conditioning. These data demonstrate the distinction between the neural mechanisms underlying the neuroendocrine responses to fear and to novel stimuli, suggesting that noradrenergic neurons are selectively involved in the hypothalamo-hypophysial responses to fear stimuli. PMID- 8725000 TI - Contribution of plasma membrane and endoplasmic reticulum Ca(2+)-ATPases to the synaptosomal [Ca2+]i increase during oxidative stress. AB - In the present study we analyzed the effect of ascorbate (0.8 mM)/Fe2+ (2.5 microM)-induced membrane lipid peroxidation on the levels of intracellular free calcium,[Ca2+]i and on the possible mechanisms involved in the perturbation of intracellular calcium homeostasis during oxidative stress. For this purpose, the influence of the ascorbate/iron oxidant system on the plasma membrane and endoplasmic reticulum Ca(2+)-dependent ATPases of brain cortical synaptosomes was studied. In addition, the influence of the peroxidative process on the uptake of calcium (45Ca2+) and on the Na+/Ca2+ exchange activity at the plasma membrane was evaluated. After ascorbate/Fe(2+)-induced membrane lipid peroxidation of the order of 18.05 +/- 4.20 nmol TBARS/mg protein, an increase in [Ca2+]i occurred, under basal or depolarizing conditions (30 mM KCl), which was dependent on the extracellular calcium concentration. Thus, for 1 and 3 mM extracellular calcium concentration, an increase of the resting [Ca2+]i values of 19.8% and 33.7% was observed, while after the K(+)-depolarization the enhancement of the [Ca2+]i was 18.4% and 29.5%, respectively. The Na+/Ca2+ exchange activity and the time dependent influx of 45Ca2+ observed in basal conditions and after the 30 mM K(+) depolarization, were not affected under the peroxidative conditions. The Ca(2+) dependent ATPase activity of the synaptosomal plasma membrane was significantly depressed following peroxidation of membrane lipids, decreasing the V(max) by 48.1%, without significant changes in the affinity of the enzyme for calcium (K(m) for Ca2+ was 0.54 +/- 0.04 microM in control conditions and 0.56 +/- 0.034 microM in peroxidized conditions). The Ca(2+)-ATPase activity of the endoplasmic reticulum was also affected during ascorbate/iron-induced oxidative stress; thus, an inhibition of 45.2% was observed 5 min after adding ATP. These data suggest that the increase in synaptosomal [Ca2+]i due to oxidative stress may result from the inhibition of the plasma membrane and the endoplasmic reticulum membrane Ca(2+)-ATPase activities, probably as a result of the alteration of the lipid environment required for the maximal activity of these membrane enzymes. The consequent increase in [Ca2+]i may be responsible for the injury of the nervous tissue observed during several pathological conditions in which free radical generation seems to be involved. PMID- 8725001 TI - Immunocytochemical localization of polyamines in the tiger salamander retina. AB - The polyamines spermine and spermidine are present in neural tissue, but their functions there are not well understood. Recent work suggests that the NMDA subtype of glutamate receptors, other glutamate receptor subtypes, and certain K(+)-channels, are neural targets for polyamines. To better understand the neuron specific roles of polyamines, we have developed antibodies that interact with spermine and spermidine in aldehyde-fixed tissue and used these antibodies in immunocytochemical studies to determine the cellular localization of these polyamines in the tiger salamander retina. The affinity-purified, polyclonal antibodies were highly specific for spermine and spermidine, exhibiting < 1% cross reactivity with putrescine, and virtually no cross-reactivity with GABA, arginine, lysine, or glutaraldehyde. Polyamine labeling was most abundant in cells in the inner half of the inner nuclear layer and in the ganglion cell layer. Some cells in the outer half of the inner nuclear layer are labeled, and there was some labeling in both synaptic layers. Double-labeling experiments indicated (1) all GABAergic amacrine cells were polyamine-positive; and (2) all ganglion cells (identified by back-filling after microinjections of rhodamine in the optic nerve) were polyamine-positive. These results are consistent with a role for polyamines as modulators of NMDA receptor function and channel function in the inner retina. PMID- 8725002 TI - Amygdala lesions block the effect of cocaine on memory in mice. AB - Post-training cocaine intraperitoneal administration enhanced memory consolidation in unoperated and sham-lesioned mice tested in a one-trial inhibitory avoidance task. Bilateral electrolytical amygdala lesions blocked this effect. The results are interpreted in terms of involvement of neurotransmitter, and in particular dopaminergic, systems in the effects of cocaine on memory. PMID- 8725003 TI - Gene expression of ND4, a subunit of complex I of oxidative phosphorylation in mitochondria, is decreased in temporal cortex of brains of Alzheimer's disease patients. AB - Gene expression of mitochondrial DNA-encoded ND4 in brains of Alzheimer's disease (AD) patients and age-matched controls was measured using Northern blot. The level of ND4 message in temporal cortex of control subjects was higher than in motor cortex, whereas the level of ND4 gene expression in temporal cortex of AD brains was decreased compared with that in temporal cortex of controls. A control probe showed no difference in expression between the two areas of AD and control brains. These and previous data suggest that neurons vulnerable to AD express higher levels of enzymes of oxidative phosphorylation than do spared neurons, and that this difference may promote selective neuronal vulnerability of AD. PMID- 8725004 TI - Ibogaine-like effects of noribogaine in rats. AB - Ibogaine is a naturally occurring alkaloid that has been claimed to be effective in treating addiction to opioids and stimulants; a single dose is claimed to be effective for 6 months. Analogously, studies in rats have demonstrated prolonged (one or more days) effects of ibogaine on morphine and cocaine self administration even though ibogaine is mostly eliminated from the body in several hours. These observations have suggested that a metabolite may mediate some of the effects of ibogaine. Recently, noribogaine was identified as a metabolite of ibogaine. Accordingly, the present study sought to determine, in rats, whether noribogaine had pharmacological effects mimicking those of ibogaine. Noribogaine (40 mg/kg) was found to decrease morphine and cocaine self-administration, reduce the locomotor stimulant effect of morphine, and decrease extracellular levels of dopamine in the nucleus accumbens and striatum. All of these effects were similar to effects previously observed with ibogaine (40 mg/kg); however, noribogaine did not induce any ibogaine-like tremors. The results suggest that noribogaine may be a mediator of ibogaine's putative anti-addictive effects. PMID- 8725005 TI - Molecular cloning and characterization of the pyrB gene of Lactobacillus leichmannii encoding aspartate transcarbamylase. AB - The Lactobacillus leichmannii pyrB gene, encoding pyrimidine biosynthetic enzyme aspartate transcarbamylase (ATCase), was cloned from a partial genomic library lying on a 1468 bp Sa/I/BstXI fragment. The predicted polypeptide sequence extending over 351 amino acid residues (M(r) 39 855 Da) was compared to those of various other organisms revealing clear identities towards them and important conservative stretches, implying that these proteins are closely related. Transcriptional initiation was mapped by primer extension and occurred 54 bp upstream of the pyrB open reading frame (ORF). Northern blot analysis indicates that the pyrB gene is transcribed as a single mRNA and not together with the following overlapping pyrC gene as a bicistronic mRNA. At high copy number the pyrB gene of L leichmannii seems to be lethal for its E coli host; inserted in a low copy vector it complements the uracil auxotrophy of an E coli pyrB mutant which shows distinct ATCase activity in the cell extract. With an excess of uracil in the growth medium the gene is apparently repressed and no ATCase activity can be measured. PMID- 8725006 TI - Production, analysis and bioactivity of recombinant vasoactive intestinal peptide analogs. AB - Recombinant vasoactive intestinal polypeptide (VIP) analogs were expressed in Escherichia coli as a fusion protein containing tandemly repeated multiple copies of a synthetic VIP gene joined to glutathione S-transferase. The encoded protein contains VIP units separated by a linker peptide, potentially excisable by a double cleavage with endoprotease factor Xa and hydroxylamine. Expression of different polyVIP genes, from 1 to 32 units, was detected and the production of a 16 VIP polymer was performed. MonoVIP analogs appended by 5 or 10 amino acids at their C terminus were released by factor Xa from this polymerized product. They were then submitted to hydroxylamine cleavage to remove the linker sequence to finally obtain a recombinant VIP analog devoid of any amino acid extension. The biological activity of the recombinant polyVIP and VIP analogs was tested. Although less efficient than the natural neuropeptide, some of these components bound to VIP receptor, activated adenylate cyclase in human colonic adenocarcinoma cells and displayed a relaxation activity on guinea pig tracheal rings. PMID- 8725007 TI - Role of adenine nucleotides in the activation of microsomal cholesterol ester hydrolase by fructose or adenosine in rat hepatocytes. AB - In the present study we have analysed the potential relationship between the cellular level of adenine nucleotides and the activity of microsomal cholesterol ester hydrolase by treating rat hepatocyte suspensions with fructose or adenosine. Fructose raised the microsomal hydrolysis of cholesteryl esters as a function of the dose. This ketose led to marked decreases in the cell level of ADP, ATP and total adenine nucleotide whereas that of AMP increased slightly, thus giving a rise in the cellular AMP/ATP ratio. The effects remained virtually constant over a period of 60 min. Incubation of hepatocytes in a Ca(2+)-free medium with or without ethylene glycol-bis(beta-aminoethyl ether) N,N,N',N' tetraacetic acid blocked by 40% the fructose-induced activation of cholesterol esterase whereas the rise in AMP/ATP was unaffected. Adenosine caused dose dependent activations of cholesterol ester hydrolase and raised AMP, ADP and ATP concentrations as well as the AMP/ATP ratio. 2-Chloro-adenosine and N6-[L-2 phenyl-isopropyl] adenosine, non-metabolizable analogues of adenosine, did not mimic the effects of the nucleoside. A positive linear correlation exists between the percentage rises in the activity of microsomal cholesterol ester hydrolase and those in the intracellular AMP/ATP ratio in fructose- or adenosine-treated cells. These results indicate that, in microsomes from intact hepatocytes, the breakdown of cholesteryl esters to yield cholesterol and fatty acids is stimulated by fructose and adenosine and this can be explained in part by the increase in the cellular AMP/ATP ratio. In the case of fructose, also a Ca(2+) dependent mechanism is involved. PMID- 8725008 TI - Calcium-dependent oligonucleotide cellular uptake. AB - Oligodeoxynucleotides (ONs) are currently being tested in clinical trials as anti viral and thrombolytic agents. Although ONs are biologically active in almost every cell line examined, these molecules are inefficiently internalized. In addition, the mechanism of ON transport to the active site in either the cytoplasm or nucleus remains ill-defined. Thus, elucidation of the mechanism of ON cellular uptake is necessary for the rational design of optimization of ON cellular uptake, delivery, and targeting. Previous studies in this laboratory have examined the mechanism of ON cellular uptake using a phosphodiester ON which is both biologically active and internalized by the Rauscher Red 5-1.5 mouse erythroleukemia cell line. The uptake is calcium-dependent since uptake is significantly reduced in calcium-free media and enhanced up to 20-fold when incubated in elevated calcium concentrations by a mechanism which may differ from uptake in physiologic calcium. Therefore, the goal of this paper is to further examine the mechanism of calcium-dependent phosphodiester ON uptake in Rauscher cells. First, to determine whether changes in intracellular free calcium [Ca]i may influence ON uptake, [Ca]i was either increased or decreased prior to addition of the ON. Second, to determine whether protein kinase C or calmodulin are involved in uptake, the activity of each of these proteins was either increased or decreased. Neither altering [Ca]i or the activity of protein kinase C or calmodulin altered the extent of ON uptake in these cells. Finally, the effect of other cations were tested for their ability to increase ON uptake. Lanthanum and cadmium increased uptake whereas sodium and magnesium had no effect. These findings suggest that cation-mediated ON cellular uptake may not result from alteration of a calcium-dependent biological process. PMID- 8725009 TI - Histone H4 stimulates glucose uptake through the insulin receptor. AB - Histone H4 stimulates the uptake of glucose in rat adipocytes and muscle cells. However, the mechanism of this unusual activity is not known. Therefore, we have begun to investigate the mechanism by which histone H4 stimulates the glucose uptake in rat adipocytes. We report that histone H4 requires 15-20 min to achieve its maximum effect and its time course is virtually indistinguishable from the time course of insulin itself. Reduction of the concentration of insulin receptors on the surface of adipocytes, either by trypsin digestion of the receptor, or by insulin-induced down regulation of the receptor, reduced the histone H4 effect as well as the insulin effects. Also, quercetin, a bioflavenoid that inhibits the insulin receptor tyrosine kinase activity, inhibits the actions of both histone H4 and insulin. However, histone H4 activity is somewhat more resistant to these interventions than insulin activity. In contrast to the activity of insulin, histone H4 does not appear to be able to down regulate the insulin receptor, since the pretreatment of adipocytes with histone H4 did not affect the subsequent actions of either insulin or histone H4. Finally, Scatchard analysis of the binding of 125I-insulin in the presence and absence of histone H4 increases the specific binding of insulin in a concentration dependent fashion. Histone H2b, a histone that does not have insulin-like activity, does not affect insulin binding. Taken together, these data suggest that the greatest portion of the insulin-like activity of histone H4 is initiated at the insulin receptor. However, the interaction of histone H4 and the insulin receptor is more complex than a simple binding of H4 to the insulin binding site. These studies may provide additional insight into alternate mechanisms for activation of the insulin receptor. PMID- 8725010 TI - Purification and characterization of acetylcholinesterase from desert cobra (Walterinnesia aegyptia) venom. AB - Acetylcholinesterase (AChE) has been identified and purified from the venom of desert cobra (W aegyptia) to apparent homogeneity using a TSK G 3000 SW gel filtration column and a Mono Q anion-exchange column. AChE was purified to homogeneity as established by sodium dodecylsulfate/polyacrylamide gel electrophoresis. The specific activity of AChE was 357 IU/mg with acetylthiocholine iodide as substrate. The denatured W aegyptia venom AChE displayed a molecular mass of 67000 +/- 3000 Da suggesting it was a single polypeptide. Isoelectric focusing of AChE revealed that the enzyme exists in different isoforms, with isoelectric points ranging between pH 7.4-7.9. The kinetic parameters (Km and Vmax) and IC50 of AChE inhibition by procaine, tetracaine and physostigmine were investigated in the present study. PMID- 8725011 TI - The ribosome as affinity matrix': efficient purification scheme for translation factors. AB - A convenient method to purify each of the non-ribosomal proteins required to translate a native mRNA in vitro is described. In this scheme, the ribosome is used as an 'affinity' matrix to selectively elute the non-ribosomal proteins required for translation that are bound to these particles. Different sets of these proteins can be eluted with solutions of Mg2+ and NH4+ of various concentrations from either 70S, or 30S and 50S particles. A scheme for the purification of each initiation, elongation and release factor and 20 aminoacyl tRNA synthetases is described. Specific examples of the purification of the initiation (IF-1, IF-2, IF-3) and elongation (EF-Tu and EF-G) factors and for a protein called 'rescue', which affects the association of native ribosomal subunits, are given. A scheme for the purification of EF-P, which stimulates peptide-bond synthesis and one of the W proteins, which permit reconstitution of translation is also described. The procedure markedly simplifies the isolation, in homogeneous form, of all the non-ribosomal proteins required to reconstruct translation. PMID- 8725012 TI - Modification of horseradish peroxidase induced by hydroxyl radicals. The influence of oxygen. AB - Reactions of hydroxyl radicals with horseradish peroxidase (HRP) have been studied by means of pulse radiolysis technique in the absence and presence of oxygen. Hydroxyl radicals, produced in excess towards enzyme, react exclusively with the protein part of HRP with the rate constant k = 1.1 x 10(11) M-1 s-1. Activity loss induced by OH. is connected with such an enzyme modification that causes both the interference with substrate binding and partial blocking of the channel used by peroxide. It is shown that in the presence of oxygen the loss of activity is ca 10% higher, mainly due to restrictions in the formation of compound I, ie ferryl [Fe(IV) = O] pi-radical cation. PMID- 8725013 TI - Do thinner rigid gas permeable contact lenses provide superior initial comfort? AB - To test the hypothesis that thinner rigid gas permeable (RGP) contact lenses provide superior initial comfort, RGP lenses of 3 center thicknesses, 0.08, 0.12, and 0.16 mm, in otherwise matched parameters, were worn for 30 min by 17 unadapted subjects in a controlled, double masked, randomized study. Comfort ratings (0 to 100) after 30 min of wear were 42 +/- 30, 55 +/- 27, and 57 +/- 28 for the 0.08-, 0.12-, and 0.16-mm thick lenses, respectively (p = 0.04, multiple analysis of variance (MANOVA)), and 93 +/- 13 for the hydrogel control lens. The thinnest RGP lens was significantly less comfortable than its thicker counterparts (p = 0.03, Univariate F-test). There were no differences among the test lenses in edge shape, front surface wettability, or static lens fittings, and small differences in movement, vertical decentration, and front surface geometry were not significantly related to comfort. We hypothesize that the greater flexibility of the thinnest lens resulted in greater deformation of the lens during a blink cycle, causing transient peripheral lens lifting and interaction with the upper eyelid, thereby reducing comfort. In conclusion we found that thinner RGP lenses do not provide an initial comfort advantage and that very thin and, as a corollary, very flexible, RGP lenses can actually be less comfortable initially than stiffer, but otherwise matched, designs. PMID- 8725014 TI - Effects of lens parameter variation on rigid gas-permeable lens adherence. AB - The influence of lens parameter variation on the frequency and persistence of rigid gas-permeable (RGP) lens adherence after overnight lens wear was investigated. In 3 separate studies, 11 subjects wore lenses of different designs in each eye over 5 nights. No significant differences in the frequency of lens adherence were found with loose vs. tight peripheral fittings (49% vs. 42%), or with 9.6-mm vs. 8.7-mm diameter lenses (42% vs. 52%). However, a significantly higher frequency of adherence was found for flat-fitting lenses compared to steep fitting lenses (84% vs. 49%; p < 0.05, Binomial test). Adherence was also significantly more persistent after eye opening with flat-fitting lenses, whereas steep-fitting lenses regained mobility more rapidly than the other lens designs tested. These findings suggest that the "suction cup" theory of RGP lens adherence does not adequately explain this phenomenon, and that the lens parameter variations investigated in this study do not reliably reduce or eliminate lens adherence during RGP extended wear. PMID- 8725015 TI - Risk of infection from sleeping with contact lenses on: causes of risk. AB - Although the risks associated with extended wear contact lenses are well-known, there has not been an adequate explanation of why sleeping with the lenses on increases the risk of infection. It is contended that the reduction in the available oxygen caused by contact lenses does not in itself explain the high rate of corneal infection caused by wearing the lenses while sleeping. This paper examines the contribution that the mechanism of sleep itself makes to the risk of infection, based on patient studies in two optometric practices. The results show that the risk of ocular infection, particularly corneal ulcers, is substantially increased with overnight wear of lenses. A review of the role of oxygen deprivation, atmospheric pollution, bacterial survival strategies, infection resistant defenses during wakefulness, and patterns of sleep indicates that oxygen deprivation of the cornea is not the only factor in the increased risk associated with wearing the lenses while sleeping. It is suggested that the combination of the introduction of airborne pollutants to the contact lens during the day, the reduced oxygen transmission through contact lenses and closed lids, and the lack of eye and lid movement during sleep together create the conditions in which bacterial infection is most likely to occur. PMID- 8725016 TI - Contact lens transmissibility: effects on delivery of oxygen to the cornea. AB - The purpose of this study was to determine the effects of rigid contact lens materials of various permeabilities and identical design on the oxygen shortfall of the human cornea and to explore differences in oxygen delivery to the cornea associated with contact lens materials of different modulus of elasticity values. Measurement of corneal oxygen uptake was performed for six human subjects under three conditions: (1) normal open eye; (2) after the static wear of six materials; and (3) after the dynamic wear of the same six materials. The 6 materials had Dk values that ranged from 0.02 to 127 (cm2/s)(ml O2/ml x mm Hg) and modulus of elasticity values from 1300 to 2200 MPa. Major results include: (1) that the oxygen shortfall of the cornea decreased with increasing lens transmissibility under both static and dynamic conditions, with significant differences (p < 0.001) being demonstrated among materials; and (2) significantly larger differences (p < 0.001) between the static and dynamic condition data were seen for materials of low Dk but were not predictable by lens modulus (p = 0.43, r2 = 0.018). The corneal demand differences between high and low Dk materials are possibly related to changes in corneal oxygen dynamics rather than to differences in tear exchange. PMID- 8725017 TI - Oxygen transmissibility at various locations in hydrogel toric prism-ballasted contact lenses. AB - PURPOSE: Corneal metabolism is known to be adversely affected by anterior surface hypoxia. It has been demonstrated that contact lens (CL) oxygen transmissibility (Dk/L in cmmL O2/s mL mm Hg) is a direct function of the oxygen permeability (Dk in cm2 ml O2/mm Hg mL mm Hg) of the plastic divided by the thickness of the lens (L in cm). It has also been shown recently that the cornea should be sensitive to the Dk/L of that portion of a CL immediately overlying the tissue, in the absence of tear exchange and mixing; such is the case for all hydrogel CLs. Because hydrogel toric CLs are known to have anisomorphically variable thickness profiles, we have measured lens thickness along the vertical meridian and calculated local oxygen transmissibilities (Dk/L) for a series of hydrogel CLs. METHODS: Thickness was measured at 5 locations along the vertical meridian of 16 different prescription toric hydrogel CLs of each of 6 brands (5 prism-ballasted design and 1 double-slab off design) and 12 different prescription spherical CLs of 2 brands (controls). Dk (at 38 degrees C) was calculated for each material from the nominal water content by a known relation. RESULTS: We found that the average Dk/L for all lenses was greatest from the center (mean +/- SD of 8 +/- 4 x 10(-9)) to the superior portion (12 +/- 2 x 10(-9) at 6 mm from the CL center and 13 +/- 4 x 10(-9) at 3 mm from the CL center) of prism-ballast design toric CLs, and least in the lower portions (4 +/- 2 x 10(-9) at 6 mm from the lens center and 6 +/- 1 x 10(-9) at 3 mm from the CL center) of these CLs. Analysis of covariance showed that the effect of location for the prism-ballasted CLs is statistically significant (F = 203.11, p < 0.00005). One nonprism ballast toric CL design and the spherical designs showed minimal differences across the vertical meridian. We therefore conclude that clinicians should carefully monitor the physiological hypoxic response of the cornea, in particular the inferior area, during wear of prism-ballasted hydrogel lenses. PMID- 8725018 TI - Effects of hydrogel lens transmissibility profiles on local corneal swelling during eye closure. AB - PURPOSE: To develop a unified model for describing the relations among local and regional contact lens transmissibility, topographical corneal location, and lens induced corneal swelling under the closed eye. METHODS: Thirteen adult male subjects, all adapted to daily wear of hydrogel lenses, wore a series of hydrogel lenses under patched eyes for 4 h. Corneal swelling was determined using an optical pachometer for five sites along the vertical meridian of test corneas. Four of the test conditions involved approximately uniform (no lens or parallel lenses) oxygen barriers over the central 8 mm of the cornea. Four additional sessions involved test lenses of -6, -3, +3, and +6 D, all having a center thickness of 0.2 mm. RESULTS: For the uniform barrier conditions, corneal swelling followed a similar pattern at all sites. Swelling was greatest centrally and least at the most peripheral, 7 mm, chord. The swelling vs. lens thickness response could be closely described by a third-order polynomial. Swelling with nonuniform thickness lenses at all sites showed evidence of small thickness averaging effects. Average central corneal swelling under the nonuniform lenses could be estimated from the polynomial function by averaging lens thickness over 5.5 to 6.75 mm for minus lenses and approximately 7.75 mm for plus lenses. CONCLUSIONS: The corneal swelling response to closed eye wear of hydrogel lenses follows a well ordered course over the central 7 mm. Over the same topographical area, the degree of swelling is influenced by both local and regional lens thickness profiles. PMID- 8725019 TI - Effects of hypoxia and hypercapnia on contact lens-induced corneal acidosis. AB - It has been assumed that contact lens wear (CLW) may induce stromal acidosis, which is a result of corneal hypoxia and the accumulation of CO2 (hypercapnia) at the tear-lens interface. However, it has not been directly shown whether hypoxia and hypercapnia are the only causes of CL-Induced corneal acidification. In this study, we provide preliminary data about the relative contributions of hypercapnia and hypoxia to CL-induced stromal acidification by monitoring pH while the cornea was exposed to a hyperbaric oxygen atmosphere. This paradigm minimized if not eliminated the pH effects of lens-induced hypoxia on all but one subject without altering the pH effect of hypercapnia. Seven subjects were fitted with hydrogel lenses; 5 with low O2 transmissibility (Dk/LO2 = 14.0 x 10(-9) (cm/s) (ml O2/[ml x mm Hg)]), and 2 with medium O2 transmissibility (Dk/LO2 = 17.2 x 10(-9) (cm/s) (ml O2/[ml x mm Hg])) lenses. After lens insertion, modified goggles were fitted to control the corneal environment by exposing 1 eye to 20%O2 and 80%N2 (air), and the contralateral eye to 80%O2 and 20%N2 (hyperbaric O2). Corneal thickness (CT) was measured before CL insertion and over 120 min of wear. We assumed that corneal hypoxia was present if CT increased during the test period. Stromal pH was measured using a slitlamp fluorophotometer before lens insertion and at 20-min intervals for a total of 80 min. After 80 min of wearing the low Dk/L lens under hyperbaric exposure, 4 of 5 subjects showed reduced pH (mean delta pH = 0.23 +/- 0.05) and no increase in CT, suggesting that only hypercapnia was contributing to acidosis. For the same lens, but with exposure to air, 4 of 5 subjects showed a larger drop in pH (mean delta pH = 0.62 +/- 0.48) compared to hyperbaric exposure and an increase in CT, indicating that both hypoxia and hypercapnia reduced pH. Subjects wearing the medium Dk/L lens showed a small but equal drop in pH under both air and hyperbaric conditions without changes in CT, suggesting that only hypercapnia was contributing to acidosis. These preliminary results suggest that both mechanisms contribute to the pH shift accompanying CLW and that the contribution of hypercapnia is approximately 30%. Finally, the effects of hypercapnia and hypoxia are dependent on individual metabolic requirements and the transmissibility of the lens to O2 and CO2. PMID- 8725020 TI - Recovery from induced corneal edema and endothelial morphology after long-term PMMA contact lens wear. AB - PURPOSE: To study the effect of long-term polymethyl methacrylate (PMMA) contact lens wear (CLW) on corneal endothelial morphology and corneal hydration control (CHC). METHODS: Sixteen long-term PMMA CL wearers (mean duration of wear 20.8 +/- 8.39 SD years; mean age 40.37 years +/- 10.47 SD) served as subjects and 14 age matched, non-lens wearing subjects (mean age 41.29 years +/- 10.15 SD) served as controls. Corneal function was assessed by measuring the corneal thickness recovery rate (PRPH) after induced edema from a hypoxic stress load. Endothelial morphology was assessed by taking specular microscopy photographs, digitizing the images, and calculating the cell density, size, and coefficient of variation of cell size (CV). RESULTS: Morphologic analysis showed a significant decrease in mean endothelial cell density (2147 vs. 2865, p < 0.01), an increase in mean cell size (582 vs. 381, p = 0.001), and an increase in polymegethism (CV, 0.32 vs. 0.26, p < 0.01) for PMMA lens wearers compared to agematched controls. Corneal function as measured by PRPH was not significantly different for PMMA lens wearers compared to controls when taken as a whole. However, a subgroup of PMMA lens wearers (seven eyes, four subjects) did show a substantial reduction in PRPH. These subjects also showed a strong relation between reduced PRPH, morphology, and years of lens wear (r2 = 0.99). CONCLUSIONS: Longterm PMMA CLW leads to increased polymegethism in most, if not all, cases and results in a substantial decrease in corneal function in some cases. PMID- 8725021 TI - Tear film lipid layer formation: implications for contact lens wear. AB - An overview of the relation between lipid layer formation and contact lens wear is provided, with particular emphasis on factors that influence the delivery and spreading of lipid on the tear film. The disruption of the lipid layer by a contact lens is discussed, as well as factors that may enhance the stability of the prelens tear film lipid layer. PMID- 8725022 TI - Characteristics of corneal staining in hydrogel contact lens wearers. AB - Corneal fluorescein staining is widely used in clinical practice and research, but little information exists on the distribution of staining in a large group of asymptomatic contact lens wearers. This cross-sectional study took place at 3 centers, and investigated the pattern of corneal fluorescein staining in both eyes of 98 hydrogel contact lens wearers. We also investigated the strategies used by three experienced clinicians to grade corneal staining. Overall corneal staining was graded using a scale from 0 to 4 in one-half steps, and five corneal zones, superior, inferior, nasal, temporal, and central, were also graded. The average overall staining grade for both eyes of our subjects was 0.50, with an average of 0.57 for the right eye and 0.44 for the left. This difference between the eyes was statistically significant (p = 0.011). In addition, a comparison of the zones within each eye showed a significant difference (p = 0.0001) among the zones. Corneal staining between the two eyes was also positively and significantly correlated (r = 0.58; p = 0.0001). Grading strategies among clinical investigators were significantly different (p = 0.0001), indicating a potential source of bias in multi-centered studies. The difference in corneal staining between the eyes may represent a source of systematic bias, and could be due to grading the right eye before the left. The correlation in corneal staining between the eyes indicates that the two eyes of a subject cannot serve as independent data points. One-third of the subjects who participated in this study had notable corneal staining. This finding underlines the importance of regularly checking corneal staining in clinical practice. PMID- 8725023 TI - Anterior hydrogel lens deposits: polished vs. unpolished surfaces. AB - Traditionally, lathe cut hydrogel lenses receive both back and front surface polishing to eliminate any surface irregularities that might serve as potential attachment sites for lens deposits or even microorganisms. However, with the advent of newer more technologically advanced lathing equipment, the need for lens polishing may require renewed justification. This study sought to compare, by scanning electron microscopy (SEM), anterior lens surface deposit formation on polished and unpolished 55% water hydrogel lenses. A total of 10 subjects wore in 1 eye (control) a lens with both front and back lens surfaces polished and on the other eye (test) a lens with only the back lens surface polished for a period of 2 weeks on a daily wear basis. Each lens was then scanned (using SEM), with 3 separate regions photographed at 30x using a 10 kV electron source. Using a Wilcoxon Signed-Ranks test on the differences between average deposit ratings for the polished and the unpolished lenses, by subject pair, we failed to reject the null hypothesis that the difference was zero. PMID- 8725024 TI - Bacterial assay of contact lens wearers. AB - PURPOSE: The goal of the project was to determine the quantity of bacteria on the contact lens and adjacent areas of the eye. This paper is a quantitative study of the contact lens and ocular aerobic microbiota in a mixed group of daily and extended wear disposable contact lens users. METHODS: The contact lens, the lower fornix, tears collecting at the lower fornix, and edge of the lower lid at the Meibomian gland margin were assayed for the quantity of bacterial colony forming units (CFU). Eighteen patients wearing 49 disposable high water content hydrogel contact lenses were assayed and the mean lens age was 8.8 +/- 4.6 days. Three patients wore their lenses on a daily wear basis and 15 on an extended wear schedule. Tear samples were obtained with sterile microbial loops and the lens was macerated into small particles with a tissue grinder. The samples were poured onto the surface of chocolate agar plates and incubated at 35 degrees C for 48 h in 5% Co2. RESULTS: The lid margin revealed the greatest bacterial presence (mean = 9.7 CFU; median = 2 CFU; mode = 0 CFU). The lens showed the next greatest presence of CFU (mean = 4.5 CFU; median = 1 CFU; mode = 0). The fornix and tears revealed the least bacterial presence (fornix: mean = 2.6 CFU; median = 0 CFU; mode = 0 CFU). The bacteria were coagulase-negative staphylococci. CONCLUSIONS: The bacterial assay of disposable lens wearing contact lens subjects indicates that the lid margins are the greatest source of bacteria with the tears being the lowest. These studies support the concept that in the eye, the lens typically does not possess a large number of bacteria under normal conditions. PMID- 8725025 TI - Relation between contact lens wear and Meibomian gland dysfunction. AB - This study attempted to determine whether contact lens wear has any adverse effect on the Meibomian glands. The study also tried to elucidate the prevalence of Meibomian gland dysfunction in the general population. The results of the study showed that Meibomian gland dysfunction exists in 43% of the population (lens wearers and nonlens wearers), 49% of the contact lens wearing population (81 subjects), and 39% of nonlens wearers (150 subjects). No statistically significant difference was found in the prevalence of Meibomian gland dysfunction between healthy contact lens wearers and the control group of nonlens wearers. The study therefore could not prove that contact lens wear is a contributing factor to Meibomian gland dysfunction. PMID- 8725026 TI - Palpebral aperture sizes of rigid and soft contact lens wearers compared with nonwearers. AB - Previous studies have shown that contact lens wear may affect palpebral aperture size (PAS). In this study, 74 wearers of rigid lenses were matched for sex and age with soft lens wearers and nonwearers. Partial face photographs were taken of all subjects, in the case of contact lens wearers, after lens removal. PAS was determined by measuring the resulting 35-mm transparencies on a Mitutoya profile projector. The mean PAS of the rigid lens wearers (9.76 +/- 0.99 mm) was found to be significantly smaller than that of the soft lens wearers (10.24 +/- 0.94 mm) and the nonwearers (10.10 +/- 1.11 mm) (p = 0.0154, analysis of variance). There was no significant difference between the PAS of soft lens wearers and nonwearers. This study supports the hypothesis that rigid lens wear causes a decrease in PAS. PMID- 8725027 TI - Centration mechanism of soft contact lenses. AB - A theoretical model is developed in which the force that restores a soft hydrogel lens to its centered position is ascribed to flexure of the lens. It is shown that flexure makes the dominant contribution to the strain energy in the displaced position. The restoring force is calculated explicitly. The return speed is estimated explicitly, assuming that the dominant resisting force is the viscous drag offered by the postlens tear film. Satisfactory agreement with observations is found. PMID- 8725029 TI - Changing patient and stone features for shock wave lithotripsy (SWL) in Turkey. AB - Shock wave lithotripsy (SWL) has made a revolution in the treatment of urolithiasis. Recent reports suggesting that stone features for SWL have changed during the last years have forced us to compare our initial and last 250 patients treated at our ESWL unit in terms of stone and patient characteristics. We found that the number of ureteric stones and small calyceal stones have increased significantly with time whereas the number of larger stones undergoing SWL has decreased significantly. We believe that this change in stone features is caused by the changing trends in the treatment of stone disease by incorporating other therapeutic options and modifying the SWL indications according to patient characteristics We also believe that prophylactic lithotripsy for asymptomatic calyceal stones still remains to be a debatable issue. PMID- 8725028 TI - A new procedure to evaluate the inhibitory capacity of calcium oxalate crystallization in whole urine. AB - A procedure to evaluate the global inhibitory capacity of heterogeneous nucleation for a given urine without any modification of its composition was developed. The procedure is based on the use of flat solid substrates that are submerged in urine. The quantification of the amount of formed crystals on the substrate permits to evaluate the crystallization inhibitory capacity for a whole urine. The proposed method was applied to study the inhibitory capacity of urines belonging to stone-formers and healthy people, before and after the ingestion of a dietary complement containing phytate in order to prove its reliability. A clear increase of the inhibitory capacity of the urine was detected in most cases when the dietary complement was consumed. Moreover, lack of crystallization inhibitory capacity was more frequent in stone-formers than in healthy people. PMID- 8725030 TI - Enzymatic evaluation of renal damage caused by different therapeutic procedures for kidney stone disease. AB - In an attempt to evaluate renal injury relative to open surgery, percutaneous nephrolithotripsy (PCN) and extracorporeal shock wave lithotripsy (ESWL) were studied in 52 patients with renal calculus disease. Preoperative and postoperative urinary levels of N-acetyl-glycosaminidase (NAG), a sensitive marker of renal tubular damage, were studied. No significant changes were noted in posttreatment urinary NAG values among patients who underwent ESWL or PCN. Although statistically nonsignificant, a constant mild increase of urinary NAG was observed after PCN, that has to be evaluated with long-term follow-up studies. The shock wave number or power in cases treated with ESWL as well as the number of renal punctures in the PCN group did not change the effect on renal tubular function. Diabetics and patients with chronic renal disease treated by ESWL did not show any significant change in posttreatment urinary NAG levels. In contrast to that, all patients treated by open surgery had significant, intense and prolonged increase of the postoperative NAG values, especially those treated by ischaemic nephrolithotomy. Comparing the three different therapeutic modalities, open surgical procedures had the most significant effect on renal function and this difference was statistically significant. We therefore suggest that ESWL does not endanger renal function, while open surgery must be reserved for selected cases. PMID- 8725031 TI - Abnormal function of renal tubules in patients with simple renal cysts. AB - The present study aimed to assess the function of proximal and distal tubules in patients with simple renal cysts. Thirty-one patients with simple renal cysts and 10 healthy subjects were examined. Based on the cyst fluid/plasma sodium ratio, 25 cysts were found to be of proximal origin and 6 of undetermined origin. In all patients cyst fluid and plasma concentrations of beta-2-microglobulin (beta-2 MG), sodium, potassium, calcium, phosphorus and total protein were assessed. Urinary excretions of sodium, potassium, calcium, phosphorus, beta-2-MG and Tamm Horsfall protein (THP) were also estimated. Fractional excretion of beta-2-MG was calculated. The concentrations of beta-2-MG in fluid obtained from cysts of proximal origin were significantly higher than in fluid from cysts of undetermined origin (2.26 +/- 0 36 vs. 0.65 +/- 0.13 mg/l, p = 0.0004). Concentrations of THP (6.85 +/- 1.21 vs. 3.14 +/- 1.06 micrograms/ml, p < 0.05), and potassium (4.39 +/- 0.07 vs. 3.13 +/- 0.44 mmol/l, p < 0.05) were also higher in fluid from proximal cysts than in fluid from cysts of undetermined origin. Plasma beta-2-MG concentration was significantly higher in patients with proximal and undetermined cysts than in the control group (4.35 +/- 0.34 and 4.11 +/- 0.74 vs. 1.89 +/- 0.1 mg/l, p < 0.001). Urinary beta-2-MG excretion was also significantly higher in both groups of patients than in healthy subjects (474.8 +/- 165.9 and 346 +/- 94 vs. 100.2 +/- 19.6 micrograms/24 h, p < 0.05). Urinary THP excretion was significantly higher in patients with proximal cysts than in healthy subjects (31.0 +/- 5.1 vs. 16.3 +/- 2.5 mg/24 h, p < 0.05). From the results obtained in this study it seems that patients with simple renal cysts of proximal origin are characterized by abnormal tubular handling of beta-2-MG, and calcium and THP excretion. Thus, in patients with proximal cysts presence of a latent renal tubulopathy seems to be likely. PMID- 8725032 TI - A complex cystic dysplastic urogenital anomaly--an odd presentation. AB - We report a case of an ectopic ureter opening into a distorted cystic seminal vesicle, with ipsilateral renal aplasia and an unusual presentation. Literature relevant to its embryology and management is discussed. PMID- 8725033 TI - Primary retroperitoneal liposarcoma: a case report and review of the literature. AB - We report a case of a 43 years old woman with a retroperitoneal liposarcoma located in the left iliac fossa. The condition was discovered because of the association of left flank pain, abdominal distention and persistent urinary infection with symptoms of vesical irritation. The diagnosis was suggested by intravenous pyelography (IVP), abdominal and pelvic ultrasonography (USG) and computed tomography (CT). Treatment was exclusively surgical. During a six-month follow-up, no recurrence was observed. PMID- 8725034 TI - Agenesis of bladder associated with multiple organ anomalies. AB - Agenesis of the bladder is an extremely rare congenital anomaly, which involves the development of urogenital sinus. Renal agenesis and some other congenital malformations are often associated. We report a 12-year-old girl with absence of bladder and solitary pelvic ectopic kidney together with some other systemic abnormalities. PMID- 8725035 TI - Appendicovesical fistula. AB - In this study, a case of appendicovesical fistula is introduced, a really uncommon complication of acute appendicitis. This special case joins the other 102 cases which have already been described in the international literature. Due to the mild and ambiguous symptomatology, the diagnosis of this kind of fistula is difficult to be confirmed, although a serious suspicion of a vesicointestinal communication does exist. Exploratory laparotomy is usually diagnostic and leads to decisive therapy. A brief review of this clinical entity is also included. PMID- 8725037 TI - Transurethral incision of the bladder neck: an objective and subjective evaluation of its efficacy. AB - This study is designed to evaluate the efficacy of treatment of bladder neck obstruction using objective (urodynamic) and subjective (assessment of satisfaction) parameters and to investigate sexual function. The results of transurethral 4 and 8 o'clock incisions of the bladder neck in 62 men (mean age 48.5 years) were reviewed with a mean follow-up of 6-12 months. Preoperative urodynamic evaluation was compared to recent postoperative urodynamic evaluation. There is marked symptomatic improvement after transurethral incision of the bladder outlet. The mean peak urine flow rate increased from 7.2 ml/s to 16.8 ml/s. Fifty-three men (85.4%) reported long-term improvement after transurethral incision of the bladder neck with an overall satisfaction rate of 71% (range 0 to 100). Five men (8%) reported new retrograde ejaculation after transurethral incision of the bladder neck. These results demonstrate that in well selected, properly diagnosed cases 4 and 8 o'clock transurethral incision of the bladder neck is an effective procedure for long-term relief of bladder outlet obstruction. PMID- 8725036 TI - Successful treatment of vesicoureteric reflux by subureteric injection of silicone. AB - Although the technique of subureteric injection has been widely accepted as an alternative to reimplantation in the treatment of vesicoureteric reflux (VUR), the choice of the material to be used is controversial. We report our experience with the endoscopic treatment of VUR by a submucosal injection of silicone. A 24 year-old man who had VUR (Grade II) and renal insufficiency was treated by endoscopic injection of silicone. Cystography after the treatment showed absence of reflux. PMID- 8725038 TI - Evaluation of a low-dose intravesical bacillus Calmette-Guerin (Tokyo strain) therapy for superficial bladder cancer. AB - To examine whether intravesical instillation of low-dose bacillus Calmette-Guerin (BCG) therapy is effective with low toxicity, we reviewed data for 111 patients with superficial bladder cancer (stages Ta and T1). Among them, 74 received the BCG treatment for prophylaxis of intravesical recurrence after transurethral resection, and the remaining 37 therapeutically for Ta or T1 tumours. The patients were divided into two groups by instillation dose of BCG (Tokyo 172 strain): 40 mg (n = 55) and 80 mg (n = 56), and statistically compared for recurrence, antitumour effect and toxicity. The mean instillations were done 8.4 times in the 40 mg dose group and 8.6 times in the 80 mg dose group. Among the 74 patients with BCG therapy for prophylaxis those with a previous episode of bladder cancer treatment (n = 47) experienced a significantly (p = 0.006) shorter recurrence-free interval than those with no episode (n = 27). Among 47 patients with a previous treatment episode, those receiving the 80 mg dose demonstrated a significantly longer recurrence-free interval than those given the 40 mg dose (p = 0.03). Among the 27 patients without previous treatment no significant difference in recurrence-free intervals was found between the two dose groups. Univariate and multivariate analyses using Cox's proportional hazards model confirmed the above findings. The recurrence index was also significantly reduced after BCG therapy in the 80 mg and 40 mg groups and similar antitumour effects for Ta or T1 tumours were observed in the two dose groups. The degree of toxicity due to BCG therapy was significantly (p = 0.02) lower with the 40 mg dose. The present study suggests that (1) a 40 mg BCG low-dose (Tokyo 172 strain) regimen is useful for preventing recurrence, with sufficient therapeutic efficacy and low frequency of toxicity, among patients without a previous treatment, and (2) prophylactic effects with the 80 mg dose regimen are much superior for previous treatment cases. PMID- 8725039 TI - Radiofrequency capacitive hyperthermia combined with irradiation or chemotherapy for patients with invasive bladder cancers. AB - To assess the immediate therapeutic clinical efficacy and long-term outcome, hyperthermia in combination with irradiation or chemotherapeutic agent was used in 46 patients with invasive bladder cancer. Radiohyperthermia (RH) was employed in 19 cases and chemohyperthermia (CH) in 27 cases. Complete response (CR) was obtained in 5 and partial response (PR) in 15 of the 46 cases. One-year survival rates by the Kaplan-Meier method were 65.0% in the CR/PR group and 46.2% in the no change (NC)/progressive disease (PD) group. Five-year survival rates were 43.8% and 18.3%, showing no difference in survival rate between the CR/PR group and the NC/PD group The overall median survival period for the CR/PR group without metastasis was 61.6 months compared to 32.3 months for the NC/PD group without metastasis (P < 0.05). PMID- 8725040 TI - Collision metastasis of bladder and prostate carcinoma to a single pelvic lymph node. AB - Genitourinary organs are at greater risk for multiple primary malignant neoplasms because of the high incidence of these tumours as primaries among all other organs. We present a case of prostate and bladder carcinoma metastasized to a single pelvic lymph node, called a "collision" metastasis, in a patient with four primary tumours. PMID- 8725041 TI - Conservative management of advanced bladder cancer. AB - Between January 1991 and October 1993, 32 consecutive patients with documented primary bladder tumours invading muscle received 3 cycles of methotrexate, vinblastin, doxorubicin and cisplatin (MVAC). The disease was re-staged by bimanual examination with the patient under anaesthesia, CT scanning and transurethral biopsy or resection. Of the 32 patients 2 underwent total or partial cystectomy and 30 did not, because re-staging showed no residual tumour in 8 (25%), stage T1-2 in 12 (37.5%) and far-advanced tumour in 10 (31.2%). The median follow-up was 2.8 years. Twelve patients with stage T1-2 tumour have required TUR, and cystectomy has not been necessary. Two patients who underwent total/partial cystectomy were all downstaged pathologically. Of the 10 failures 5 patients died of disease and 5 are alive with metastatic disease. The overall survival rate was 84.3% (27 of 32) and was 96.8% for patients with a functioning bladder. The data suggested that this active regimen can clinically induce downstaging in a significant number of patients with primary muscle-infiltrating bladder tumours. Transurethral resection plus MVAC chemotherapy is important for increased curability in patients with advanced bladder cancer. PMID- 8725042 TI - Blood loss during transurethral prostatic resection with continuous bladder irrigation. AB - A total of 120 men with benign prostatic hypertrophy were assigned randomly into three groups. Transurethral prostatic resections were performed in all patients. In Group I the operation was performed with continuous flow bladder irrigation, in Group II with suprapubic flow irrigation (without suction pump) and in Group III without continuous flow irrigation. Blood loss was determined by a colorimetric method. There were no statistically significant changes in blood loss between the groups of patients. PMID- 8725043 TI - Prostate-specific antigen density: the role in benign prostate hyperplasia, prostate intraepithelial neoplasm, organ-confined prostate carcinoma and advanced prostate carcinoma. AB - To determine the relative role of prostate-specific antigen density (PSAD) in the early detection of prostate cancer and to assess the hypothesis that PSAD offers significant advantages over prostate-specific antigen (PSA) alone in the evaluation of patients with benign (BPH), pre-malignant (PIN) and malignant prostatic diseases, we studied retrospectively 149 patients who were evaluated with either prostatic biopsies or by surgical means. Mean PSAD was calculated to be 0.1 for BPH patients; 0.09 for PIN-1 patients; 0.1 for PIN-2 patients; 0.51 for organ-confined prostatic carcinoma (CaP) patients and 1.7 for advanced CaP patients. Although we could not be able to differentiate BPH from PIN-1 and PIN-2 by using PSAD alone (p > 0.05), there were statistically significant differences between BPH versus localized CaP, PIN-2 versus localized CaP and localized CaP versus advanced CaP (p < 0.05). In conclusion we suggest that the information provided by PSAD is superior to absolute PSA values in the differentiation between BPH and CaP but PSAD was not able to add more information on differentiating BPH from pre-malignant conditions. PMID- 8725044 TI - Relationship between prostate-specific antigen, prostate volume, retention volume and age in benign prostatic hypertrophy (BPH). AB - The relationship between plasma PSA, prostatic adenoma mass (AM), retention volume (RV) and age was studied in 112 men with benign prostatic hypertrophy (BPH). Their age ranged from 48 to 85 years. Mean PSA was 6.9 ng/ml (SD 6.6) and did not exceed 10 ng/ml in 81.2% of the patients. Mean AM and RV were 66.2 g (SD 36) and 51.7 ml (SD 1.3), respectively. Increase in PSA level was influenced mainly by RV (r = 0.53) but was found to correlate weakly with AM (r = 0.34) which in turn showed low correlation with RV (r = 0.32). Age did not correlate with PSA, AM and RV (r > or = 0.1). AM and RV influence PSA levels independently and large inter-patient differences are well characterized by SD values. PMID- 8725045 TI - Changes in serum levels of prostatic acid phosphatase and prostate specific antigen after luteinizing hormone-releasing hormone analogue administration in patients with metastatic prostatic cancer in relation to glandular differentiation. AB - Eighteen previously untreated patients with metastatic carcinoma of the prostate were treated with LHRH analogue. They were divided into 3 groups according to the degree of glandular differentiation. In all groups, a transient rise of PAP and PSA was observed after the LH and testosterone surge. However, relative values of LH, testosterone, PAP and PSA did not differ significantly among the 3 groups. These facts suggest that a transient rise of PAP and PSA is caused by testosterone surge independently from the degree of glandular differentiation after LHRH analogue administration in patients with advanced prostatic cancer. PMID- 8725046 TI - Skene's duct cyst in adult women: report of two cases. AB - Skene's duct cysts in the adult are uncommon lesions. Complete urologic examinations are necessary because these lesions simulate other clinically important lesions. Herein we report two cases of Skene's duct cyst in adult women. In these cases we could make differential diagnosis by physical, radiological and endoscopic examinations. PMID- 8725047 TI - Evaluation of impotent men with intracorporeal injection of papaverine and colour Doppler ultrasound. AB - Fifty impotent patients have been evaluated with intracorporeal injection of papaverine and colour Doppler ultrasound. The papaverine tests were negative in 34 of 50 patients. Nineteen of 34 patients with negative papaverine results showed normal colour Doppler findings. In 14 of 16 patients with positive papaverine results the colour Doppler analysis revealed normal structure and functions. Correlation between intracorporeal papaverine injection and colour Doppler ultrasound results was found in 59% (p < 0.01). We concluded that intracorporeal papaverine injection is an effective, simple diagnostic test for beginning evaluation of impotent men. However, this screening test should be supported by colour Doppler ultrasound. PMID- 8725048 TI - Enhanced glomerular phospholipase activity in the obstructed kidney. AB - Previous study demonstrated that an increment of glomerular eicosanoid production may contribute to the haemodynamic changes in the obstructed kidney. To elucidate the mechanisms responsible for enhanced glomerular eicosanoid production, the present study was designed to investigate activities of related enzymes by isolated glomeruli from rat kidney with unilateral ureteral obstruction (UUO) or bilateral ureteral obstruction (BUO) for 24 hours. The activity of phospholipase A2 (PLA2) was determined by monitoring 14C] arachidonate release using 14C] phosphatidylcholine (PC) or 14C] phosphatidylethanolamine (PE) as a substrate. Phospholipase C (PLC) activity was assayed by measuring the release of [3H] inositol triphosphate [3H] IP3 from [3H] phosphatidylinositol 4,5 biphosphate ([3H] IP2). The activity of PE-specific PLA2 was increased in glomeruli from the kidney with BUO and the contralateral kidney of unilateral ureteral obstruction (CLK). PLC activity was significantly greater in the cytosolic fraction of glomeruli from kidneys with UUO, BUO and CLK compared to sham-operated control. The activity of PC-specific PLA2 was not significantly increased in any group. These results indicate that the increased synthesis of eicosanoids by glomeruli from obstructed kidney may be mediated by enhanced activities of PE-specific PLA2 and PLC. The increased activities of these phospholipases by glomeruli from CLK may contribute to a compensatory response. PMID- 8725049 TI - Light scattering of human skin: a comparison between zinc (II)-phthalocyanine and photofrin II. AB - Zinc(II)-phthalocyanine is the active component of the liposomal formulation CGP 55847 which showed a highly activity in photodynamic therapy studies on a variety of animal tumours (K. Schieweck et al., SPIE Conf. Proc., 2078 (1994) 107-118). The photophysical properties of zinc(II)-phthalocyanine have been studied in detail and compared with those of Photofrin II(R), the only sensitizing agent approved so far for Phase III and IV clinical trials (M. Ochsner-Bruderer, Inaugural Dissertation, University of Basle, 1994). As will be shown in a series of papers, the main photophysical properties of zinc(II)-phthalocyanine are significantly better than those of Photofrin II(R) (M. Ochsner-Bruderer, Inaugural Dissertation, University of Basle, 1994). In this paper we especially consider the effect of the absorption wavelength on the penetration of light into the human skin. The results clearly show that the longer absorption wavelength of zinc(II)-phthalocyanine causes a deeper penetration of light into the human skin as compared with Photofrin II(R). In addition to this, the higher extinction coefficient (epsilon S) lowers the zinc(II)-phthalocyanine dose required to induce a tumour necrosis. PMID- 8725050 TI - Merocyanine 540 mediated photoirradiation of leukemic cells. In vitro inference on cell survival. AB - In order to evaluate the selective killing of merocyanine 540 (MC 540) mediated photoirradiation in neoplastic cells, bone narrow cells from children with leukaemia or neuroblastoma and normal children as well as peripheral blood cells and Reh-6 and HL-60 cell lines were studied. Cell suspensions were incubated with MC 540 and exposed to various argon laser 514 nm doses. Cell survival was estimated with trypan blue supravital stain following a 24 h incubation and has been followed in continuous cell cultures of 4 weeks duration. Our results showed that the inhibition of survival of neoplastic haemopoietic cells by laser in the presence of MC 540 is proportional to the MC 540 and photoirradiation doses. A 99.9999% inhibition of Reh-6 and HL-60 was noted at irradiation doses where the corresponding mean survival of normal bone narrow cells was (33.6 +/- 15.5)% and (50.6 +/- 10.7)% respectively. Peripheral blood mononuclear cells were not sensitive to MC 540 mediated photoirradiation. The inhibition of survival of bone marrow metastatic neuroblastoma cells was (69.9 +/- 4.1)%. In conclusion, it seems that MC 540 mediated photoirradiation in neoplastic cells exerts selective cytotoxicity and can be used in ex vivo purging of malignant cells in the bone marrow. PMID- 8725051 TI - Photochemical formation of singlet molecular oxygen (1O2) in illuminated aqueous solutions of p-aminobenzoic acid (PABA). AB - Evidence is presented for the photochemical formation of singlet molecular oxygen (1O2) in air-saturated buffered aqueous solutions of p-aminobenzoic acid (PABA) using sunlight-range illumination. This is significant because PABA is widely used as an active ingredient in sunscreen preparations that are applied to the surface of the skin and 1O2 is known to cause oxidative damage to cells via the formation and subsequent reactions of lipid peroxides. Furfuryl alcohol (FFA), a well known chemical trap for 1O2, was added to aqueous PABA solutions prior to illumination. The FFA was consumed when the solution was illuminated, but no loss of FFA occurred in the dark and loss by direct photolysis was negligibly slow. Further evidence for the formation of 1O2 in illuminated aqueous PABA solutions is provided by the results of experiments in which individual solutions containing PABA and FFA that were diluted with D2O exhibited an increased rate of FFA consumption due to the increased lifetime and concentration of 1O2 in this solvent. PMID- 8725052 TI - Induction and repair of DNA damage in UV-irradiated human lymphocytes. Spectral differences and repair kinetics. AB - The alkaline elution assay has been employed to study the induction and repair kinetics of DNA damage in human lymphocytes after irradiation with biologically relevant doses of UVB (297 and 302 nm) or UVA (365 nm) radiation. At 365 nm, when the predominant lesions are single-strand breaks, the rate of lesion induction was 1.5 x 10(-3) per 10(8) Da per kJ m-2. The number of breaks decayed with a half-life of about 50 min after a dose of 20 kJ m-2. In the UVB region, cyclobutyl pyrimidine dimers and 6-4 photoproducts are formed, both of which are repairable via the nucleotide excision repair pathway. By using repair inhibitors, the rate of induction of such lesions at 297 and 302 nm was found to be 0.07 per 10(8) Da per J m-2. Lesions were removed with a half-life of about 100 min. Mathematical modelling of the excision repair process revealed a time dependent polymerization-ligation rate: after an initial lag phase the polymerization-ligation rate increased, reaching 50% of its maximum rate at 80 100 min after the start of repair incubation. This course of development might be due to a damage-associated regulation of DNA precursors synthesis. PMID- 8725053 TI - Electron paramagnetic resonance evidence of the generation of superoxide (O2.-) and hydroxyl (.OH) radicals by irradiation of a new photodynamic therapy photosensitizer, Victoria Blue BO. AB - Electron paramagnetic resonance (EPR) experiments were performed on Victoria Blue BO, a cationic dye whose photocytotoxicity has been studied against the human leukaemic cell lines K-562 and TF-1. EPR experiments with 2,2,6,6-tetramethyl-4 piperidone and spin trapping with 5,5-dimethyl-1-pyrroline-N-oxide showed that, on illumination in aerated aqueous solution or DL-alpha dipalmitoylphosphatidylcholine liposomes, photoexcited Victoria Blue BO is unable to generate 1O2, whereas O2.- and .OH are trapped by 5,5-dimethyl-1-pyrroline-N oxide in the presence or absence of electron donors. The O2.- formed probably leads to the .OH radical, with an efficiency which is increased by electron donors such as FE2+. PMID- 8725054 TI - Ex-vivo treatment of gastric Helicobacter infection by photodynamic therapy. AB - Attempts to develop PDT for eradication of Helicobacter infection have only been successful in vitro. We have investigated the effect of topical sensitization (except ALA) of Helicobacter mustelae on explanted ferret gastric mucosa using one of five sensitizers (methylene blue (MB), toluidine blue O (TBO), phthalocyanine, haematoporphyrin derivative and 5-aminolavulinic acid), followed by irradiation with an appropriately tuned copper vapour pumped dye laser. A 90% reduction in counts of bacteria sensitized with 0.75 mg TBO kg-1 were seen after irradiation with 200 J cm-2. Concentrations of MB of 0.75 mg kg-1 and 7.5 mg kg-1 were not toxic to H. mustelae, but the further addition of 20 J cm-2 laser light reduced colony counts by more than 99%. MB at a concentration of 75 mg kg-1 exhibited significant dark toxicity towards H. mustelae, but further addition of 20 J cm-2 laser light resulted in near eradication of all colonies. The remaining three compounds were ineffective. Finally, we studied the microscopic fluorescence distribution of MB (7.5 mg kg-1) on ferret gastric mucosa after topical administration. Fluorescence was greatest in the superficial mucosal layer, upon which lies the bacteria. However, from experiments on rats, the energy required to kill the sensitized bacteria was insufficient to damage the underlying mucosa. We conclude that Helicobacter can be killed on host mucosal epithelium following topical administration of MB and subsequent exposure to laser light. PMID- 8725055 TI - Characterization of pyridoxal phosphate as an optical label for measuring electrostatic potentials in proteins. AB - This paper presents data which allow one to characterize pyridoxal 5'-phosphate (PLP) as an optical label for electrostatic potential measurements in proteins. Experimental studies were carried out with 6-N-(5'-phosphopyridoxyl)-2-N-acetyl lysyl methyl ester (PLP-ALME) as a model compound which simulates PLP covalently bound to a protein. Calculations of electrostatic potential maps were done using the model compound 2,4-dimethyl-3-hydroxy-5-hydroxymethyl phosphate-pyridine (DHHPP). Studies on relative changes in PLP-ALME fluorescence and absorbance vs. pH resulted in four pKa values of PLP-ALME in solution which can be used as intrinsic pKa values (pKint) of the ionizable groups of the label covalently bound to the protein. The pKa values obtained from fluorescence data are 4.1, 5.6, 8.7 and 11.1 and those from absorbance data are 3.1, 4.7, 8.7 and 11.0. The differences between corresponding pKa values are related to differences in PLP ALME behaviour between the excited and ground electronic states and to intramolecular charge-charge interactions. Quenching of PLP-ALME fluorescence by I-, acrylamide and Cs+ at pH 6, 7 and 8 shows that in the case of I- and acrylamide the Stern-Volmer constants of quenching (Ksv) decrease with increasing pH, while the opposite is true for Ksv of Cs+. These results as well as the analysis of electrostatic potential maps of DHHPP show that with its ampholytic character PLP can be used to measure local electrostatic potentials in the pH range 5-9. PMID- 8725056 TI - In vitro UVB irradiation of bovine crystalline lens causes cell damage and reduction in leucine aminopeptidase activity in lens epithelium. AB - Past studies in our laboratory have shown that low levels of UVB can cause changes in the optical properties of organ cultured ocular lenses, while other research has shown that in vitro UV radiation causes decreases in leucine aminopeptidase activity in homogenates of crystalline lens material. Therefore we have investigated whether there is a relationship between such decreases in enzyme activity and changes in lens optics and structure. Organ cultured bovine lenses were irradiated with low doses of UVB, and lens optics, histology and leucine aminopeptidase activity (leucine beta-naphthylamide hydrolysis at pH 7.5) were assessed daily. Lenses irradiated with 0.1 J cm-2 UVB showed a decrease of about 30% in leucine aminopeptidase activity 1 h after irradiation, while changes in lens optics were not observed until at least 24 h after irradiation. Histological examination of the lens anterior epithelium revealed changes in epithelial cells ranging from pyknotic nuclei to large areas of cell fragmentation. The results of this study suggest that a decrease in soluble aminopeptidase activity in lens epithelial cells may be a direct result of the epithelial cell damage rather than an effect of UVB on the enzyme per se. PMID- 8725057 TI - Adjacent guanines as preferred sites for strand breaks in plasmid DNA irradiated with 193 nm and 248 nm UV laser light. AB - 193 nm UV laser light induces single strand breaks as well as double strand breaks in plasmid DNA. The frequency of strand breaks is increased at sites where at least two guanine or, less frequently, a guanine and an adenine are adjacent to each other. 248 nm UV laser light induces predominantly single strand breaks with a less pronounced preference for guanine clusters. At both wavelengths, the presence of oxygen does not change the pattern of strand breaks, but in the presence of nitrous oxide, selectivity is lost; this is attributable to the production of the hydroxyl radical. These findings can be explained by a model in which the propagation of a radical or an electron hole along the DNA helix competes kinetically with the strand cleavage reaction. The difference in selectivity at the two different wavelengths is ascribed to the preferential light absorption by the purine bases at 193 nm. PMID- 8725058 TI - Effects of sonodynamic and photodynamic treatment on cellular thiol levels. AB - Exposure of murine leukemia L1210 cells to ultrasound in vitro resulted in cell fragmentation; cells which survived this treatment were 100% viable and showed a transient decrease in intracellular thiol pools (expressed as glutathione equivalents). Depletion of thiol pools became progressively greater with increasing exposure of cells to ultrasound; this was not associated with leakage of thiols into the medium. Biochemically elevating glutathione levels or sonication in the presence of the anti-oxidant trolox offered only a minor degree of protection against sonotoxicity. Photodynamic therapy also resulted in depletion of intracellular thiol levels, and loss of cell viability. In the presence of trolox, both effects were reversed, although elevation of intracellular thiol levels did not protect cell from phototoxicity. PMID- 8725059 TI - [The role of adjuvant therapy in the management of pN2 non-small-cell carcinoma of the lung]. AB - Over a period of eleven years (1983-1993), the role of adjuvant chemo and/or radiotherapy was evaluated on 222 resected patients (pts) with NSCLC at atage IIIA(N2). All the patients underwent an attentive mediastinal limphoadenectomy. Fifty-five patients had a clinical mediastinal node involvement. 174 pts had a single mediastinal node station involved while 48 had two or more stations involved. One hundred and seventy-one pts (77%) underwent adjuvant therapies, consisting of citotoxic chemotherapy in 40 pts, radioterhapy in 97 pts and chemoradioterhapy in 34 pts. Follow-up lasted until September 1994. Overall 5-yr survival was 17.5%, the median being 17 months. Forty-two pts were, at that moment, still living (median 43.5 months, min 11-max 120) with 37 disease free. We verified a significant difference concerning survival among the three histologic types (p = 0.03), with the squamous achieving the best result (21.3% at 5-yrs). Surgical N2 had a better survival (20/5% at 5-yrs) than the clinical one (9%), (p = 0.01). In particular, if only one nodal station was involved, survival was 21.3% compared to 4.5% when metastases were present at two or more nodal station (p = 0.0001). Considering the level of mediastinal node involvement, the worst prognosis was linked to the carina node metastases (p = 0.02). Survival benefits were obtained by means of adjuvant therapies (20/2% vs 8.1%), (p = 0.0002). Analyzing all the pts, the best survival was achieved in those treated by surgery plus chemo-radiotherapy (32.4%), (p = 0.0001). As regards the squamous cell tumors, pts who underwent surgery plus radiotherapy had the best prognosis (24.2%), (p = 0.0026). Further, in non-squamous cell tumor, chemo-radiotherapy increased survival (45%), (p = 0.0001). At multivariate analysis, only the level of nodal involvement and the adjuvant therapy maintained their statistical significance. Our results prompt us to conclude that: 1) Squamous cell tumors and single nodal station involvement are linked to the best prognosis; 2) Squamous cell carcinoma should be treated by adjuvant radiotherapy; 3) Non-squamous tumors should undergo to chemo-radiotherapy; 4) Clinical N2 (according to our recent experience) may benefit from neoadjuvant chemotherapy. PMID- 8725060 TI - [The role of surgery in the management of diffuse (N3) lung cancer]. AB - The aim of the paper is to give a contribution to the understanding of the role of surgery in patients with N3 lung cancer. The A. describe their results on 32 patients with N3 lung cancer operated on mediastinectomy; histological examination of the specimens showed 27 squamous cell carcinomas and 5 adenocarcinomas. The operation was performed through a median sternotomy to gain access to the left and right paratracheal nodes, pretracheal, precarinal and subcarinal nodes, subaortic and left and right hilar nodes that were removed together with the fat of the mediastinum. Left pneumonectomy was performed in 5 patients. There was no operative mortality; 12 patients were irradiated in the postoperative period with 5,000 Rads (50 Gy), while 18 or the 27 patients with squamous cell carcinoma are still alive. PMID- 8725061 TI - [Tracheal sleeve lobectomy in the radical surgery for lung cancer]. AB - Tracheal sleeve lobectomy is right upper lobectomy extended to include resection of the main bronchus, the carina and a segment of the trachea with end-to-end anastomosi of the trachea and end-to-side anstomosis of the intermediate bronchus on the main left bronchus, or the upper lobe on the trachea. This operation is possible with the aid of high frequency jet ventilation. The surgical indication of this operation is bronchoscopic and functional and guarantees a good oncological radicality in the presence of N0. PMID- 8725062 TI - [Blood preservation in thoracic surgery: feasibility and limitations]. AB - The transfusion of homologous blood carries risks associated with reactions to transfusion and the transmission of diseases. Minimizing surgery-related blood use is therefore of utmost importance. However, the application of blood-saving methods in surgery is restricted by a number of factors. Thoracic surgery poses particular problems in this regard due to the type of diseases treated, patients' general status and the use of neoadjuvant chemotherapy protocols. The aim of this study was to analyse the feasibility of a blood conservation program for thoracic surgery. A detailed analysis of blood-sparing techniques was carried out and the factors influencing blood consumption were evaluated in the preoperative, intraoperative and postoperative periods. Although pre-operative autologus blood donation is an extremely important factor, its effect is limited in thoracic surgery since only a few patients are suitable candidates for such a procedure. Our personal experience confirms the difficulty of applying a blood sparing program to thoracic surgery. Nevertheless the risks associated to transfusions, especially those related to the use of homologous blood, can be reduced by optimizing existing blood sparing techniques or developing new hemostatic agents. PMID- 8725063 TI - [Pleuro-pericardial cysts. Case reports and clinical-diagnostic as well as therapeutic considerations]. AB - Three patients with pleuropericardial cysts are reported. The authors accoding to what exists in literature confirm the congenital origin of this relatively rare pathology. It is benign and rapresents 6-19% of all mediastinal neoplasms. This lesion is more common in adults and prefer IV-VI decade of life. Usually this cyst constitute a casual radiological report because most of these tumours are asymptomatic. In our experience only one case was characterized by dyspnea. The authors, besides, consider CT of great help in diagnosis of pleuropericadial cyst in view of its high specificity. Thoracoscopy, result very important to diagnosis and exeresis of this neoplasm when doubts no persist as to the true nature of the lesion. Surgical treatment of pleuropericadial cyst is the only therapy. In our experience thoracotomy was preferred with excellent results and no postoperative complications or recurrences. PMID- 8725064 TI - [Extracapsular lobectomy in benign monolobar thyroid diseases]. AB - On the basis of their experience acquired in the field of thyroid surgery the authors examine the problems related to extracapsular lobectomy from a tactical and technical point of view, starting with its principal indications: benign monolobar thyroid disease in a single or multiple nodular form. The validity of extemporary histological tests is also assessed on the basis of their experience of rare false negatives and the relative successive totalization programme. Lastly, the paper underlines the fundamental identification of the recurrent nerve as the central point of the operation, before which the authors emphasize that nothing should be cut or ligated apart from the superior vascular peduncle and vena media. PMID- 8725065 TI - [Synchronous carcinoma of the colon and rectum]. AB - The incidence of synchronous carcinoma of colon-rectus is rising in relation to a greater oncogenic environmental charge and increased average life expectancy. There is also a risk of not recognising the disease (especially small carcinoma or multicentric neoplasm). After a Literature review, two clinical cases are described by the authors. They recommend an extensive use of preoperative colonoscopy and a careful intraoperative exploration of the viscera. It is also important that patients undergo periodical postoperative endoscopic controls. PMID- 8725066 TI - [Superior mesenteric artery syndrome after surgical correction of severe dorso lumbar deformities. Authors' experience]. AB - The authors report two cases of superior mesenteric artery syndrome observed in 22 operations for severe spinal deformities. Medical, dietary and postural treatment enabled the situation to be resolved in one case; whereas an intestinal derotation according to Strong-Valdoni was successfully performed in the other. The various surgical alternatives are discussed. PMID- 8725067 TI - [Pseudoaneurysm in pancreatic pseudocysts. Report of a clinical case]. AB - Pancreatitis in a percentage of 1.5% in a pseudocyst, 10% of which are represented as formations of the pseudoaneurysmatic type one of the visceral artery. The authors of the following study present the case of an 86 year old woman with positive anamnesis because of a chronic pancreatitis, who is affected by a symptomatology in the epigastric regione together with hematemesis and melena. An ecography shows at the head level a mass of cm 6.5 hypo ecogena, delimitated by a hyper ecogena band. A TAC, with and without contrast means and integrated by some RM and angio-RM sequences, shows that it is a vascular injury, with the characteristic area on enhancement after the infusion of the contrast means. The patients goes under surgical intervention, showing a trough supplied by a branch of the hepatic artery. The intervention is performed with the tying of the artery, with the drainage of the residual cavity plus the creation of a pancreatic, cystic and jejunum derivation. PMID- 8725068 TI - [Colon volvulus]. AB - The authors report a clinical case and review the international literature. After an analysis of the incidence and the predisposing factors causing this disorder, they focus attention on the question of therapy. In forms with vital loop non surgical derotation must be attempted. In the event of the failure of non invasive treatment and to prevent recidivation these forms are treated surgically, also using videolaparoscopy. In forms with non-vital loop, surgery consists of the section of the necrotic segment and preparation of anastomosis which may be immediate or deferred depending on general and local conditions. PMID- 8725069 TI - [Synchronous carcinomas of the colon and rectum]. AB - The authors base their observations on 3 cases of synchronous carcinoma of the large intestine and 1 case of association of cancer on polyps and synchronous colorectal carcinoma. After a short review of the etiopathogenetic and diagnostic aspects, they focus attention in particular on the various types on surgical approach which synchronous carcinoma of the large intestine offer to surgeons. The authors underline that numerous forms of surgery exist which are often complex and difficult, especially if multiple neoplasia involve separate colic segments and above all if they affect the distal rectal section. In conclusion, they affirm that the association of cancer on polyps and synchronous colorectal carcinoma is not rare and should be treated using combined endoscopic and surgical therapy. To the precise colic exeresis should be followed by endoscopic resection in the case of a scissil, villous polyps with high non-differentiated neoplastic tissue laying close on the endoscopic plane of section. PMID- 8725070 TI - [A case of appendiceal mucocele associated with cancer of the colon]. AB - Mucocele of the appendix is a rare lesion (only 0;2-0.3% of all appendicectomies). It is characterized by distension of the lumen within mucus. Mucoceles are often asymptomatic and are discovered as an incidental finding either at surgery or during barium edema. There have been reports of other tumors associated with adenomatous change in the appendix. The most common association was with adenocarcinoma of the colon, which occurred concurrently in approximately 20% of patients. Preoperative diagnosis is rare. The therapy is surgical; or appendicectomy in the benignant lesions of right colectomy in the malignant lesions. The authors report a case of appendiceal mucocele associated with adenocarcinoma of the colon. That was occasionally observed during emergency laparotomy in perforation of cave viscer. We discuss problem that appear from the association of these two pathologies. PMID- 8725071 TI - [Spigelian hernia: apropos of two cases]. AB - Two observed cases of spigelian hernias in 54 years and 62 years old men leads to an analysis of the diagnostic clinical and therapeutic aspects of this rare abdominal pathology. It represents 1-2% of all abdominal hernias and can occur anywhere along the semicircular line of Douglas, but in 90% of cases its most frequent localization is from the level of the umbilicus to the inguinal area. Diagnosis can be difficult because of non-specificity of symptoms. Many times Spigelian hernia is masked by abdominal fat and the only symptom is pain. TAC plays a primary role in the diagnosis of spigelian hernia, infact with it, detailed characteristiques and location of the hernia, hernial content, other intra-abdominal pathology, are recognized. The only treatment of spigelian hernia is surgical; many times it is diagnostic and therapeutic. A pararectal skin incision permits an accurate exploration of the abdominal cavity and an cautious choice of aponeurosis to the plastic repair. PMID- 8725072 TI - [Giant cavernous angioma of the liver]. AB - The authors report on their experience with a case of liver hemangioma; after exploring the trade-off between the risk and expected benefit from resection of giant liver hemangioma, because of this searched literature for studies of with rupture. Controversy persists regarding management of these lesions with the development of non invasive modalities treatment, such as use of embolizing agents; Elective surgery can be performed, such as treatment resective, according to the size of the hemangioma. PMID- 8725073 TI - [Video-laparoscopic cholecystectomy with suprapubic approach. Technical note]. AB - The authors describe a personal variant of the videolaparoscopic technique of cholecystectomy. This is characterised by the use of an open access to the abdominal cavity following the execution of pneumoperitoneum through an incision in a non-umbilical but suprapubic site, and the use of only three incisions and therefore only three operating trocars. Twenty-nine cholecystectomies have been performed using this personal technique of which only 2 had to be converted to open surgery. The reasons were not attributable to the technique and no complications were observed. The authors also performed a total of 99 laparoscopic cholecystectomies between February 1993 and September 1994 with a total of 4 reconversions and 1 complication (choleperitoneum). Concomitant lithiasis of the choledochus was present in 4 cases and treated with operative ERCP prior to laparoscopic cholecystectomy. Throughout this period only 4 cholecystectomies were performed using a laparotomic access roule owing to concomitant choledochal lithiasis and non-cannulatable papilla during ERCP. The reasons that led the authors to prefer the suprapubic access route were both aesthetic and technical owing to the reduced interference of the videocamera with other instruments, whereas the use of only 3 trocars was also based on the need to economise as well as on aesthetic grounds. PMID- 8725074 TI - [Esophago-jejunal anastomosis after total gastrectomy with circular suturing: using a personal method]. AB - The authors describe a personal technique of oesophagus-jejunum anastomosis post total gastrectomy for gastric cancer, inspired by the Oesophagel transection technique. This consists of introducing into the oesophagus the detachable head of the circular stapler by gastrotomy before sectioning the oesophagus and removing the stomach and subsequently carrying out anastomosis as usual. The principal advantage of this technique is in respect to the oesophageal margins to be anastomosed which remain immune to ischemic lesions, lacerations and retractions of the mucosa, with a minor incidence of anastomotic leakage. PMID- 8725075 TI - [Use of porous plyethylene (Medpor) in maxillofacial surgery]. AB - The use of alloplastic material in maxillofacial surgery is having a much larger diffusion in substituting missing bone parts and in incrementing those that already exist. Medpor, a substance composed of porous polyethylene, has been of relatively recent introduction in the surgical field. Due to its characteristics of biological compatibility, easy morphological adapatability and mantaining it's initial volume after beeing positioned, Medpor lends itself well as an alternative to autogenous grafts or other alloplastic materials. The authors consider twenty patients operated between May 1991 and September 1993 with Medpor grafts; seven of these were affected by microgenia, and were submitted to vertical increase of the chin after horizontal osteotomy by interposing Medpor. Two patients affected by hemifacial microsomia had been treated with grafts on the mandibular border in order to improve their facial symmetry. In three cases Medpor was used in patients affected by previous traumas: to reconstruct a fractured orbital floor with bone loss, the dorsum of the nose, and in order to regain protrusion of the cheekbone. In two patients affected by UCLP Medpor was used in sustaining the lateral region of the nose as well as in disguising hypoplasia of the upper jaw. In other two cases Medpor had been used to fill a depression created in the temporal region where the muscle had been rotated to create a flap; other uses of Medpor have been: as a filler of titanium mesh, used in the reconstruction of a nasal valve and in the camouflage of the upper jaw. The authors report complications only in three cases all involved the exposure or prominence of the Medpor grafts. In one case the prominence of the grafts which was positioned on the upper jaw appeared in the upper part of the fornix; in another case the exposure was post-traumatic and in the third case the exposure followed the dehiscence of the titanium prosthesis within which the Medpor had been positioned. Except for the last case, in which the patient died--for other reasons--healing of the graft exposure was obtained by a simple remodeling. PMID- 8725076 TI - [Salivary HIV antibodies. Review of current data and future prospectives]. AB - Medical researchers, especially those involved in AIDS studies, have shown increasing interest in the salivary diagnosis in the recent years. Saliva sample collection is simple, quick, non traumatic and it can be done at low cost. Moreover saliva specimens can be collected without significant infection risks for both patients and operators. The presence of HIV-antibodies in the saliva, along with the above mentioned reasons, makes saliva sample a promising means of testing for HIV antibodies. The attached report summarizes the results of studies carried out by various researchers to assess the reliability of several test methods for HIV-antibodies. PMID- 8725077 TI - [Iatrogenic lesions of the facial nerve during parotid surgery for pleomorphic adenoma]. AB - The authors, after a survey of the literature, describe the injuries of the facial nerve during parotid surgery for pleomorphic adenoma. The facial nerve damages are often transient with total resolution, except where one or more branches are deliberately sacrificed for malignant tumors or recurrence. PMID- 8725078 TI - [Calcifying odontogenic cyst. A clinical case]. AB - The calcifying odontogenic cyst (COC) is a rare lesion that involves the jaws which, at times, may evolve into a solid, benign, tumor-like mass. There is no significant sex predilection, but it is probably more common in later life. The diagnosis is base only on histologic examination that represents, in the best of cases, the only way to differentiate the COC from other pathologies. Such lesions include: odontogenic cyst, residual cyst, globulomaxillary cyst, lateral periodontal cyst, ameloblastoma (multicystic and unicystic), odontogenic mixoma, and still with calcifying odontogenic epithelial tumor (Pindborg tumor) and adenomatoid odontogenic tumor. The authors report a case of COC, starting point for a review of the clinical, radiographic and histomorphologic features of the lesion. PMID- 8725079 TI - [Consensus conferences: instructions for their use]. PMID- 8725080 TI - [The treatability of refractory or resistant hypertension by personalized antihypertensive chronotherapy based on ambulatory monitoring of the arterial pressure]. AB - The present paper is aimed at detecting whether the combination of ambulatory blood pressure monitoring (ABPM) and antihypertensive chronotherapy can be usefully exploited in the treatment of hypertension which appears to be rebel or resistant to pharmacological administration. The study was carried out in 28 hypertensive patients (15 men and 13 women, 45 +/- 10 years old), rebel (10 cases) or resistant (8 cases) to a conventional antihypertensive therapy. The ABPM was performed weekly in order to see when the eventual patient was cured by the therapy which was synchronized to the "relative hypertensive peak" of the antimeridian, postmeridian and nocturnal part of the day. The actuarial analysis documented that at the end of the sixth week no case remained rebel or resistant to the antihypertensive chronotherapy, suggesting that the combination of the ABPM with the chronotherapeutical treatment is highly recommended in presence of apparently refractory cases of hypertension. PMID- 8725081 TI - [Alteration of luteinizing hormone pulsatility in patients with arterial hypertension]. AB - The pattern of luteinizing hormone (LH) secretion in men with mild and moderate hypertension was studied. LH pulsatility was evaluated for eight hours in 14 male patients, who were subdivided into two groups: group A, consisting of 8 patients, whose systolic blood pressure ranged between 180-160 mmHg and the diastolic between 104-95 mmHg; group B, 6 patients whose systolic blood pressure ranged between 220 and 180 mmHg and the diastolic between 115-105 mmHg. Seven healthy adult males were evaluated as a control. The major changes of LH pulsatility in group A included an increased peak width (p < 0.05), increased peak amplitude (p < 0.001) and increased peak area (p < 0.001). In group B the changes followed the same pattern as in group A, but were more pronounced. The number of LH peaks was reduced (p < 0.01), the peak width was increased (p < 0.05), and both peak amplitude and peak area were increased as compared to the control group (p < 0.001). Our study demonstrates that the pattern of LH pulsatility is altered in essential hypertension and the main feature is represented by the prolonged duration of LH peaks and their greater amplitude. The altered pattern of LH secretion is likely to reflect a primary hypothalamic derangement with the gonadotropin releasing hormone (Gn-RH) secreting neurons remaining synchronized for longer times and secreting larger Gn-RH masses than in normal subjects. PMID- 8725082 TI - [Treatment of refractory heart failure with different dialysis technics]. AB - The efficacy of the dialysis in the heart failure non responsive to the traditional methods is well known. In our study we have evaluated the effects of three different dialytic methods (UFI: ultrafiltration isolate; HF: hemofiltration; CAVH: continuous arteriovenous hemofiltration). These methods are used to cause a good depletion in the patients with intractable heart failure following from ischaemic valvular or primitive cardiomiopathy. We have treated 18 patients (6 for each method) and the patients with organic or functional renal disease were treated with the methods (HF and CAVH) that have depletive and depurative effects. Each of these methods is capable, without important differences, of achieving the following aim: the hydro-sodium depletion, the correction of the haemodynamic alteration and the re-establishment of the response to the traditional medical treatment. Using the hemofiltration and continuous arteriovenous hemofiltration, both soft methods, we have obtained values of dehydratation, absolute and for each session, higher than isolated ultrafiltration. The follow-up has not pointed out differences, of both prognosis and survival among patients treated with the three methods studied, whose effects are always only temporary; only the heart transplantation or the valvular correction, by operating in a very important way on the basal cardiopathy, is the resolutive event. PMID- 8725083 TI - [Job syndrome (hyper-IgE) and hypo-IgA. A rare association of immunodeficiencies]. AB - Job' syndrome and IgA immunodeficiency are a rare dysfunction of the immune system. In this work, we reported a case of a young woman who had recurrent episodes of bacterial infections in the urinary tract and genital, generalized erythematous eczematous patches and stomatitis of oral mucosa and fever. During the hospitalization, laboratory data showed high immunoglobulin IgE and low IgA levels. The T-lymphocyte presented a reduction of CD8+ cells. Tests of granulocyte function have showed a global deficit in the in vitro and in vivo chemotaxis. The correlation between these two clinic conditions is not completely clarified but it is possible to hypothesize that CD8+ lymphocytes produce an inhibition factor of chemotaxis. Job' syndrome is characterized by a selective reduction of CD8+ cells subpopulation which have an immunoregulatory function on the production of IgE by plasmacells. In the ipoIgA, an intrinsic inability of B IgA cells to proliferate and to differentiate produce a defect in the IgA production. In these two clinic disorders there is an effective dysfunction of immune system. It is possible to hypothesize that an effective defect of CD8+ cells and an immaturity of B-cells may coexist in our patient. That justifies an abnormal production of Ig and a defect in granulocyte chemotaxis. PMID- 8725084 TI - [Diagnosis of neoplastic ascites: combined use of albumin gradient between serum and ascites and cholesterol in ascitic fluid]. PMID- 8725085 TI - [Health cooperation and human development]. PMID- 8725087 TI - Do general practitioners need guidelines? PMID- 8725086 TI - [Oxygen therapy and pulmonary hypertension in chronic obstructive bronchopneumopathies]. AB - Pulmonary hypertension has a negative prognostic value in the progression of chronic obstructive lung disease. The hypoxic vasoconstriction and subsequent morphological alterations that occur in the small arteries and in the pre capillary arterioles may contribute to the reduction of the pulmonary vascular bed. The evolution of pulmonary hypertension in patients with chronic obstructive lung disease is not rapid except during relapses of the disease. Oxygen therapy reduces pulmonary arterial pressure values not only via a reduction in the functional abnormalities of the vessels but also via a regression in the anatomic changes induced by hypoxemia. Thus, long-term oxygen therapy may slow the progression of pulmonary hypertension in the course of chronic obstructive lung disease and the longer is oxygen administration the better haemodynamic results are obtained. The Medical Research Council (MRC) Study and the Nocturnal Oxygen Therapy Trial (NOTT) have clearly demonstrated that mortality among hypoxiemic patients treated with low-flow oxygen therapy is reduced; the NOTT also reported a reduction of about 10% in the levels of pulmonary vascular resistances in patients treated with long-term oxygen therapy, while an increase of about 7% has been observed in patients treated with oxygen therapy only during the night. Nevertheless, the reduction of mortality observed during long-term oxygen therapy not always is accompanied by an haemodynamic improvement; in particular a reduction in the pulmonary arterial pressure seems not to reduce mortality. In conclusion long term oxygen therapy may slow the progression of pulmonary hypertension during chronic obstructive lung disease while the improvement in pulmonary haemodynamic due to oxygen therapy is not strictly correlated with a reduction of mortality. PMID- 8725088 TI - Health problems in family practice. An Icelandic multicentre study. AB - OBJECTIVE: To establish epidemiological data on the health problems within family practice in Iceland by multicentre analysis of well-defined geographic areas. DESIGN: Prospective practice audit. SUBJECTS AND SETTINGS: Thirteen Icelandic health centres (HC) with computerized contact data from 1 January - 31 December 1988. MAIN OUTCOME MEASURES: Health problems during one year in a population, as perceived by health care providers. RESULTS: A total of 176 384 health problems during one year in a population of 31 248, as perceived by the health care provider, were analysed. Musculoskeletal disorders accounted for 9.3% of all health problems (prevalence 210.6/1000 inhabitants), respiratory disorders 9.4% (189.9/1000), accidents 7.4% (203.2/1000), cardiovascular disorders 7.4% (112.0/1000) and mental disorders 6.1% (87.6/1000). The commonest single health problems were: hypertension, upper respiratory tract infections and non-articular rheumatism. The health problems accounting for the most frequent contacts were: mental disorders (4.0 contacts per individual per year), cardiovascular (3.7), and endocrine, nutrition and metabolic (3.2). CONCLUSION: Problem-oriented medical records from HCs, computerized in a uniform standardized way, can give extensive information about the content and burden of health problems in family practice and presumably public health. Our results are valuable because the population (the denominator) and the geographic study area are well defined. This information is an important part of clinical epidemiology and can be of great value for educators and health care planners. PMID- 8725089 TI - Does the doctors' professional experience reduce referral rates? Evidence from the Finnish referral study. AB - OBJECTIVE: To examine hospital referral rates in Finnish health centres according to doctors' and health centres characteristics. DESIGN: Survey of all general practice hospital referrals over one week. SETTING: Central and northern part of Finland, in a region comprising 72% of the area of the country and one-third of the population. PARTICIPANTS: 851 health centre doctors. OUTCOME MEASURES: Referral rates in terms of characteristics of doctors and health centres. RESULTS: During the study period, the 851 health centre doctors had 58 760 consultations (mean 69 patients/week), and 2 921 (5%) patients were referred to secondary care. The variation between the highest and lowest referral quintile of the doctors was almost 17-fold, and of the health centres 2.4-fold. Relatively more referrals were made by locums, young, not yet graduated and female doctors than by specialized, more experienced and male doctors. CONCLUSION: A low referral rate is closely connetected with the extent of the general practitioners' professional experience and specialist training. Investing in specialist training and continuing medical education seems to be the best way to reduce high referral rates. PMID- 8725090 TI - Smoking in pregnancy: a way to cope with excessive workload? AB - OBJECTIVE: To examine if abstention from smoking in pregnancy increases with opportunities to limit workload. DESIGN: Cross-sectional study based on self administered questionnaires to women post partum. SETTING: All maternity wards in Norway, 16 October-26 November 1989. SUBJECTS: 5 438 women, or 87.2% of all parturients in Norway during the study period. MAIN OUTCOME MEASURE: The abstention fraction (AF) defined as percentage of nonsmokers in pregnancy among prepregnancy daily smokers. RESULTS: The abstention fraction was lower for women with lack of influence on work pace, with hectic work pace, exhaustion after work or work schedules other than daytime, for women with children <16 years of age in the household, and for women in paid work with 20 hours or more of housework weekly, than for women without these conditions. Adjusted odds ratios (OR) for abstention were low for women with children <16 years both when in paid work (OR 0.4) and as housewives (OR 0.3). Women who received extra help with housework had a high abstention fraction (OR 1.6) both when in paid work and as housewives. The abstention fraction was high for women with daytime work (OR 1.5) and low with hectic work pace (OR 0.7), compared with women without these conditions. CONCLUSION: Abstention from smoking in pregnancy increases with opportunities to limit total workload. PMID- 8725091 TI - Diagnosis of maxillary sinusitis in Finnish primary care. Use of imaging techniques. AB - OBJECTIVE: To observe the effect of imaging techniques on the diagnostic pattern of sinusitis in primary care. DESIGN: A multicentre survey. SETTING: 14 health centres with varying facilities for clinical imaging. SUBJECTS: 446 adult patients with a suspicion of acute maxillary sinusitis. Ultrasound or radiography facilities existed in 337 cases while 109 could only be judged clinically. MAIN outcome measures--The use of ultrasound, radiography, laboratory tests, irrigation and control visits. The final number of patients with sinusitis in different facility groups. RESULTS: When available, ultrasound was used in 82-92% and radiography in 6-32% of cases. The ultrasound finding showed mucosal thickening or was difficult to interpret in every third case; one half of these were interpreted as sinusitis and the rest were considered healthy. In total, sinusitis was diagnosed in 84-88% when ultrasound or radiography only could be used, and in 77% when both techniques were available (p < 0.01). Simultaneously the general practitioners' confidence in the correctness of their diagnosis increased from 39% to 66%. CONCLUSION: When possible, ultrasound is widely used in diagnosing sinusitis in Finnish primary care. The use of ultrasound slightly diminished the numbers of sinusitis diagnoses but the techniques of using and interpreting ultrasound findings need to be improved. PMID- 8725093 TI - Young women with symptoms of urinary tract infection. Prevalence and diagnosis of chlamydial infection and evaluation of rapid screening of bacteriuria. AB - OBJECTIVE: To estimate the prevalence of chlamydial infection among young women with UTI symptoms. To evaluate chlamydia diagnostics with the aid of enzyme immuno assay (EIA) on first-void urine. To evaluate rapid screening of bacteriuria, including low concentrations of common pathogens. DESIGN: EIA for detection of Chlamydia trachomatis antigen and confirmation with immunofluorescence test (DFA) in urine, cervical and urethral chlamydia culture, nitrite and granulocyte esterase test, urine sediment, chamber count, dipslide and conventional urine culture were performed. SETTING: Primary health care (PHC). PATIENTS: 217 women aged 15-35 years attending PHC for dysuria or urgency frequency. MAIN OUTCOME MEASURES: Frequency of chlamydial infections. Sensitivity, specificity, predictive values of EIA and bacteriuria screening tests, respectively. RESULTS: The frequency of chlamydial infection was 3.7%. In spite of a high specificity of the EIA test (0.94 without DFA) the number of false positives exceeded the number of true positives. No single bacteriuria test showed sufficient diagnostic efficiency. CONCLUSIONS: Routine chlamydia testing in young women with UTI symptoms is recommended. EIA test on urine is of little use. Assessing diagnosis of UTI symptoms requires insight into the use of several rapid tests and a dialogue with the patient. PMID- 8725092 TI - Cancer incidence on a small island--research opportunities in general practice. AB - OBJECTIVE: To test the impression of an increased cancer incidence on the island of Pellworm (in the far North of Germany) and to illustrate the feasibility of a general practice-based approach in epidemiological research. DESIGN: Cancer incidence on Pellworm was prospectively registered in the only general practice on the island from 1986 to 1992. Age-standardized rates and expected rates were calculated on the basis of the Saarland cancer registry, the only registry in Germany. Standardized incidence ratios and 95% confidence intervals (CI) for Poisson-distributed events were also calculated. The cancer data were summed up over a seven-year period. SETTING: The only general practice on the island of Pellworm, a North Frisian marshland-island. SUBJECTS: The total practice population between 1986 and 1992 (N = 1172) RESULTS: The crude annual cancer incidence rate for Pellworm was, according to the impression, unexpectedly high: 634/100 000 for men and 502/100 000 for women. After age standardization, however, this increased rate of cancer incidence was even lower than in the Saarland (ratios: 0.86 for men and 0.95 for women). Only the incidence of neoplasms of the lymphatic and haematopoietic tissue in men exceeded the limits of statistical likelihood (ratio: 3.21; 95% CI: 1.17-7.10). CONCLUSION: The impression of an increased rate of cancer on Pellworm could not be validated. The overall incidence rate was even lower than expected. Only the rate of leukaemia/lymphoma (men) was significantly higher. Reasons for this result could not be detected by a descriptive approach. General practice is a suitable place for studies in cancer epidemiology, especially in such ideal circumstances as a clearly limited area and complete and reliable documentation. PMID- 8725094 TI - Side-effects of chiropractic spinal manipulation: types frequency, discomfort and course. AB - OBJECTIVE: To make a preliminary appraisal of the types of unpleasant reactions reported by patients after spinal manipulation and to estimate their frequency. STUDY DESIGN: Standardized interview data collected in a longitudinal survey. METHOD: Ten chiropractors collected data on all unpleasant reactions that were reported after a maximum of six visits by ten consecutive patients per chiropractor, with the use of a questionnaire that contained mainly closed-ended questions. RESULTS: Following 368 treatments in 95 patients, some type of side effect was reported after 1/3 of treatments. Local or radiating symptoms were most commonly reported (23%). No alarming events were reported. Ninety percent of all reactions were graded by the patients as moderate or slight. They commenced on the day of therapy in 87% of cases, and had disappeared within 24 hours in 83%. CONCLUSION: Results from this preliminary study indicate that reactions to spinal manipulation may be relatively common but benign in nature and of short duration. No estimate could be made of rare or serious side-effects. PMID- 8725095 TI - Who is using antihypertensive drugs? A prescription analysis from Finland. AB - OBJECTIVE: To analyse the prescribing of antihypertensives in Finland in 1993. DESIGN: A descriptive three-month follow-up study of reimbursed prescriptions for chronic hypertension dispensed in Finnish pharmacies. SETTING: The nationwide prescription data base of the Social Insurance Institution covering 80% of Finnish pharmacies. Material--The study material consisted of 479 744 antihypertensive prescriptions from ATC-groups hypotensives (C02), diuretics (C03), beta blocking agents (C07), and potassium (A12B) for 279 435 hypertensive patients. RESULTS: Of all the prescriptions (excluding potassium supplements), 30% were for beta blocking agents, 24% for diuretics, 22% for calcium channel blocking agents, 20% for ACE inhibitors or ACE inhibitor + diuretic combinations, and 4% for other hypotensives. Two thirds of the men received a drug from a hypotensive group, nearly half were prescribed a beta blocking agent, and 27% a diuretic. Among women the distribution of the different drug groups was more even: more than half the women (55%) were prescribed hypotensives while beta blocking agents and diuretics were prescribed for 43% and 44%, respectively. Due to the different treatment profile between men and women the expenses of treatment also differed. The cost of prescriptions for female patients was, on average, 17% less than that for male patients. CONCLUSION: The choice of antihypertensive drugs depends on the age and sex of a patient. Prescribing antihypertensive drugs does not fully meet national recommendations. New drugs are gaining ground in the treatment of hypertension. An increase in the cost of treatment will result from this development. PMID- 8725096 TI - Identifying patients for research in general practice. PMID- 8725097 TI - Snags in statistics and questionnaires. PMID- 8725098 TI - A continuous epitope from transmissible gastroenteritis virus S protein fused to E. coli heat-labile toxin B subunit expressed by attenuated Salmonella induces serum and secretory immunity. AB - Antigenic site D from the spike protein of transmissible gastroenteritis virus (TGEV), which is a continuous epitope critical in neutralization, has been expressed as a fusion protein with E. coli heat-labile toxin B subunit (LT-B) in attenuated S. typhimurium. Synthetic peptides containing the sequence of site D induced TGEV neutralizing antibodies when inoculated subcutaneously in both rabbits and swine. A synthetic oligonucleotide encoding residues 373-398 of TGEV S protein, including antigenic site D, was cloned in frame with the 3' end of LT B gene, into a plasmid used to transform S. typhimurium delta asd chi 3730. A collection of 6 recombinant plasmids designated pYALTB-D I-VI encoding LTB-site D fusions with a variable number of site D sequences were selected. Four of the 6 LTB-site D fusion products expressed in S. typhimurium chi 3730 formed oligomers (pentamers) that dissociated at > 70 degrees. S. typhimurium chi 3730 (pYALTB-D) V and VI expressed the oligomer forming products with higher antigenicity. Partially purified LTB-site D fusion product expressed from S. typhimurium chi 3730 (pYALTB-D) V induced anti-TGEV neutralizing antibodies in rabbits. Recombinant vaccine strain S. typhimurium delta cya delta crp delta asd chi 3987 transformed with plasmid pYALTB-D V expressed constitutively products that formed oligomers presumably containing 20 copies of site D, and showed a high stability in vitro. This recombinant strain was orally inoculated in rabbits and induced TGEV specific antibodies in both serum and intestinal secretion. PMID- 8725099 TI - The effect of transcription on genetic recombination in poxvirus-infected cells. AB - We have examined the effects of transcription on recombination frequencies in poxvirus-infected cells. A synthetic poxviral promoter was shown to function as a hybrid early/late transcription element when fused to a luciferase reporter gene, and then cloned into genetically-marked recombination substrates. These lambda DNA substrates were transfected into cells infected with Shope fibroma virus (SFV) and the recombinants detected by recovering the transfected DNA, packaging it in vitro into infectious particles, and then assaying the yield of recombinants on Escherichia coli. Controls showed that the poxviral promoter conferred no replicative advantage, or disadvantage, on molecules encoding the promoter. Furthermore, the promoter had no detectable effect on the recombination frequency when recombination was measured in the interval immediately adjacent to the promoter-insertion site. However, the promoter did appear to stimulate recombination at a distance, in a manner that appeared to be dependent on the level of transcription, and the effect was observed regardless of whether or not the promoter was present on one or both of the recombinational substrates. The peak of recombinational enhancement was centered about 500 bp away from the promoter element, where the frequency of recombination was 30-50% higher than that seen when the recombinational substrates lacked the promoter. Possible explanations for these observations are discussed. PMID- 8725100 TI - Mutations affecting the cytoplasmic domain of the Moloney murine leukemia virus envelope protein: rapid reversion during replication. AB - Five premature termination mutations and five missense mutations were introduced into the portion of cloned Moloney murine leukemia virus (M-MuLV) DNA encoding the Env cytoplasmic domain. All of the mutant DNAs gave rise to replication competent virus after transfection of NIH/3T3 cells, but several of the mutant DNAs scored as replication-defective when introduced into Rat2 cells. Cell lines stably expressing the mutant DNAs all released virion particles, and in all but one case infectious virus were generated. These viable mutants were all found to have reverted to the wild-type sequence. To generate fully mutant virus stocks, the mutant DNAs were introduced transiently into COS cells, which are resistant to infection with MuLV, thus prohibiting reversion by error-prone mechanisms involving reverse transcription. Virions harvested from the COS cells were confirmed as mutant by analyzing both virion proteins and the viral DNA they generated, and were then tested for infectivity in NIH/3T3 cells. The mutant viruses were infectious, but still rapidly gave rise to revertants. We conclude that the mutations within the cytoplasmic domain do not provide an absolute block to virus replication, but that the mutants replicate more slowly than the wild type and quickly give rise to revertants with selective advantage for replication. PMID- 8725101 TI - Identification of immunodominant, group-specific and subcomplex-specific, continuous epitopes in the core regions of Japanese encephalitis virus using synthetic peptides. AB - Two flaviviruses, Japanese encephalitis (JE) virus and Dengue (DEN) virus which have high pathogenicity for humans, continue to pose a serious public health problem in tropical and subtropical countries of the world. In order to identify the immunodominant B-cell epitopes for diagnostic application, we have prepared a series of 15-mer synthetic peptides from JE virus core protein based on computer analysis. Four linear, immunodominant epitopes corresponding to amino acids 91 105 (P78), 1-15 (P73), 8-22 (P74), and 34-48 (P75) of JE virus core proteins were identified by employing an enzyme-linked immunosorbent assay (ELISA), using high titered immune sera from JE-vaccinated children. P78 was found to be the most immunodominant. The sero-specificity of these peptides was tested by binding to seroconverted samples from JE and DEN-1 patients. P78 and P74 belonged to group specific epitopes which reacted with both JE and DEN-1 patient sera. P73 and P75 belonged to subcomplex-specific epitopes which reacted only with JE but not with DEN-1 patient sera. The study suggests that these peptides corresponding to the immunodominant epitopes of JE virus core protein might have the potential to be used as peptide-based diagnostic reagents for the detection and differentiation of JE and DEN antibody responses. PMID- 8725102 TI - Growth characteristics and protein profiles of prototype and wild-type rat coronavirus isolates grown in a cloned subline of mouse fibroblasts (L2p.176 cells). AB - Rat coronaviruses (RCVs) infect laboratory rats and confound biomedical research results. In vitro systems developed so far have limited the growth in knowledge about RCVs by not permitting generation of plaque-cloned virus stocks, reliable isolation of RCVs from rat tissues, or growth of high titered stocks of all isolates. Due to the fact that less than 20% of L2(Percy) cells were becoming infected, sublines were produced and selected for maximal growth of RCVs. Screening of 238 cell sublines yielded L2p.176 cells which were highly susceptible to all RCVs tested; however, susceptibility declined after 30 passages in vitro. Low-passaged L2p.176 cells were used to isolate virus from natural outbreaks and to propagate individual RCV plaques into high titered stocks. Proteins from six RCV isolates were immunoblotted using polyclonal rat and mouse antibodies to sialodacryoadenitis virus and polyclonal monospecific rabbit and goat antibodies against the peplomer (S) and nucleocapsid (N) proteins of mouse hepatitis virus (MHV). Proteins of two prototype, one Japanese and three wild type RCVs were examined and found to be similar to those of MHV, although the exact sizes and ratios of protein forms were unique for most RCV isolates. This study reports the development of a continuous cell line which reliably supports RCVs opening an opportunity for further in vivo studies of the biology of these agents. As a first step in the characterization of RCVs, we have shown that RCV proteins are very similar to those of MHV. PMID- 8725103 TI - Synthesis of leader RNA and editing of P mRNA during transcription by rinderpest virus. AB - Purified rinderpest virus was earlier shown to transcribe in vitro, all virus specific mRNAs with the promoter-proximal N mRNA being the most abundant. Presently, this transcription system has been shown to synthesize full length monocistronic mRNAs comparable to those made in infected cells. Small quantities of bi- and tricistronic mRNAs are also synthesized. Rinderpest virus synthesizes in vitro, a leader RNA of approximately 55 nucleotides in length. Purified rinderpest virus also exhibits RNA editing activity during the synthesis of P mRNA as shown by primer extension analysis of the mRNA products. PMID- 8725104 TI - Plasmids containing cos ends inhibit the replication of phage phi CTX in Pseudomonas aeruginosa. AB - In bacteriophage phi CTX, the cohesive end sequences cos, the integrase gene int, the attachment site attP (the target site for int) and the gene ctx encoding a pore-forming cytotoxin CTX, are clustered. Phi CTX can infect some Pseudomonas aeruginosa strains with a subsequent induction of CTX expression. The 41 and 477 bp fragments containing cos ends of phi CTX DNA were cloned into the high copy number plasmid pHA10. After pretransformation with the cos ends containing plasmids, plaque formation of phi CTX and cytotoxin production in phi CTX infected Pseudomonas aeruginosa cells decreased by 100- and 10-fold respectively. Twelve hours after phi CTX infection proteins binding with cooperativity to cos sequence containing cleavable ends and the 10 bp flanking sequences were detected by gel electrophoretic mobility retardation of [32P]cos DNA. The results suggest that the cos binding proteins of phi CTX are involved in phi CTX replication. PMID- 8725105 TI - Coxsackievirus B3 clinical isolates and murine myocarditis. AB - Fifteen clinical coxsackievirus B3 (CVB3) isolates were assessed for cardiopathologic capabilities in adolescent male CD-1 mice in comparison to two well characterized cardiovirulent CVB3 strains. One isolate was cardiovirulent, one minimally cardiovirulent and the remaining 13 isolates were noncardiovirulent. The two cardiovirulent isolates and one well characterized cardiovirulent strain, established higher viremic titers, in comparison to five noncardiovirulent isolates that were examined. The two cardiovirulent isolates and one well characterized cardiovirulent strain replicated to significantly higher titers than five noncardiovirulent isolates in primary cultures of murine neonatal or adolescent cardiac fibroblasts. Nucleotide sequence analysis of an area defined by nucleotides(N)300-N599 in the 5'-nontranslated region were performed on the two well characterized cardiovirulent CVB3 strains, the two cardiovirulent isolates and 12 noncardiovirulent isolates. The data detected a single discriminatory nucleotide position. An A was present at N565 in three of four cardiovirulent CVB3, whereas a U or C was present in this position in 12 of 12 noncardiovirulent CVB3. In toto, these data are compatible with the hypothesis that the type of the nucleotide at N565, a position within the internal ribosome entry site, is associated with capacity of a CVB3 for replication in vivo and in vitro and this capacity for vigorous replication is associated with cardiovirulence. PMID- 8725106 TI - Glycoprotein B (gB) of pseudorabies virus interacts specifically with the glycosaminoglycan heparin. AB - We have previously shown that the pseudorabies virus (PrV) glycoproteins gB and gC (former PrV-gII and PrV-gIII) exhibit heparin-binding properties. While PrV-gC functions as the major adsorption protein, the biological role of the heparin binding properties of PrV-gB are not understood. We used a gC-deleted PrV-mutant, PrV (dlg92/dltk), to analyse the heparin-binding properties of PrV-gB and the biological role of the PrV-gB-protein in adsorption. PrV-gB was the only glycoprotein of this vaccine strain binding to immobilised heparin in in vitro assays. Presence of the gC-protein was not necessary for the interaction of gB with heparin. Soluble heparin also interfered with adsorption of this mutant virus to a similar extent as it blocked adsorption of wild-type PrV (Ka), but it had only a minor inhibitory effect on infectivity of the mutant strain. These results show that PrV-gB interacts specifically with immobilized heparin and heparin-like structures on the cell surface, but this interaction is not required for a productive infection. PMID- 8725107 TI - Detection of contamination of vaccines with the reticuloendotheliosis virus by reverse transcriptase polymerase chain reaction (RT-PCR). AB - The reverse transcriptase polymerase chain reaction (RT-PCR) was applied to detect contamination of Marek's disease (MD) vaccine with reticuloendotheliosis virus (REV). The env primers were used for the 1st RT-PCR to amplify the DNA fragments of REV-A and -T. The rel and env primers were used for nested-PCR to confirm the sites deleted from REV-T and REV-A. Specific amplification products were detected in the 1st RT-PCR with these primers. By nested PCR with the env and the rel primer pairs, the products originating from REV-A and -T were identified. This system, using the env primer pairs, showed a specific amplification with several REV strains (REV-T, DE, CE, KI and 0202), but no amplified product was detected with MDV, NDV, IBV or ILTV. The 1st RT-PCR detected the virus in a concentration of 10(3) in 50% fluorescent antibody infectious dose per ml (FAID50/ml) and the nested PCR detected 10(1) FAID50/ml virus. The sensitivity of the RT-PCR system was found to be higher than that of the FA assay. This system provides a rapid, sensitive and specific method for detection of contamination of MD vaccines with REV-RNA, and it may be applied for quality control of live vaccines. PMID- 8725108 TI - Brefeldin A inhibits the antiviral action of interferon against encephalomyocarditis virus. AB - Brefeldin A (BFA), a unique fungal metabolite of a 13-membered lactone ring, exhibits various biological actions, including antitumor, antifungal and antiviral activities. In the present study, mouse LB cells were treated with various concentrations of interferon (IFN) and/or BFA overnight and infected with encephalomyocarditis virus (EMCV) after removal of IFN and BFA. Doses of BFA which neither inhibit the metabolism of the cell nor the infectivity of EMCV, decreased the IFN-induced antiviral activity against EMCV as demonstrated by virus titer from supernatants. Since 2-5A synthetase and double-stranded RNA (dsRNA)-dependent protein kinase (PKR) have been suggested to be involved in the antiviral action of IFN against EMCV, their activities were investigated in LB cells after BFA treatment. Northern blot analysis and in situ hybridization showed a decrease (2-3-fold) in the mRNA of 2'-5' oligoadenylate (2-5A) synthetase after BFA treatment. BFA also inhibited the activity of 2-5A synthetase, 2-5A dependent RNase and phosphorylation of PKR in cellular extracts, indicating that BFA may be exerting its inhibitory effect both at the transcriptional and post-transcriptional levels. This study reports a new biological action of BFA, demonstrating that BFA antagonized the antiviral action of IFN by inhibiting IFN-induced enzymatic pathways. These studies also suggest that both 2-5A and PKR are important in the antiviral activity of IFN against EMCV. PMID- 8725109 TI - Infectious in vitro transcripts from cloned cDNA of the potato A potyvirus. AB - A full-length cDNA copy of potato virus A RNA was constructed downstream from the bacteriophage T7 RNA polymerase promoter. A single extra guanosine not present in PVA RNA was added to the 5'-end of the cDNA. The capped in vitro transcripts from cloned cDNA were infectious in the mesophyll protoplasts and intact plants of Nicotiana tabacum. PVA coat protein accumulated in transcript-inoculated tobacco protoplasts and infected systematically intact plants producing high PVA titres according to the ELISA and immunoblot assay. The infected tobacco plants produced similar mild mosaic symptoms in the systematically infected leaves, irrespective of whether the RNA transcripts or parental virus were used as inoculum; the virus particles were also morphologically similar according to immunosorbent electron microscopy. PMID- 8725110 TI - Methylation of the polyomavirus major capsid protein VP1. AB - Polyomavirus VP1 has been shown to be modified by phosphorylation, sulfation, acetylation and hydroxylation. The major capsid protein VP1 is now shown to be modified by methylation. Addition of cycloheximide to infected cultures followed by addition of [3H-methyl]-L-methionine and subsequent immunoprecipitation, SDS PAGE and fluorography revealed methylation occurring on VP1. Amino acid analysis of [3H-methyl]-L-methionine-labelled polyomavirus VP1 by two-dimensional paper chromatography and HPLC of the acid-hydrolyzed protein revealed the presence of 3H-labelled trimethyllysine and monomethylarginine. PMID- 8725111 TI - The complete nucleotide sequence of RNA-2 of a fungally-transmitted UK isolate of barley mild mosaic bymovirus and identification of amino acid combinations possibly involved in fungus transmission. AB - The complete nucleotide sequence of RNA-2 of a fungally-transmitted UK isolate of barley mild mosaic bymovirus (BaMMV isolate UK-F) was determined and compared with other published sequences, particularly UK-M, an isolate derived from the same source but which has been mechanically passaged for several years, has a deletion of about 1 kb and cannot be fungally transmitted. From an alignment of the BaMMV RNA-2 encoded protein with that for barley yellow mosaic bymovirus (BaYMV), several regions of consistent homology were identified and extensive searches made for similarities with the proteins of other fungally-transmitted viruses, especially amongst the furovirus capsid readthrough proteins which seem especially prone to deletion and which have already been implicated in fungus transmission. The amino acid combinations ER (glutamic acid-arginine) or QR (glutamine-arginine) were found consistently in all of the viruses. They occurred in positions predicted to be on the outside of the protein, and therefore available for interaction with the fungus vector, and were also within the regions prone to spontaneous deletion. In view of the lack of other structural or sequence homologies, it is suggested that these motifs are strong candidates for involvement in fungus transmission. PMID- 8725112 TI - The structural protein p54 is essential for African swine fever virus viability. AB - Protein p54, one of the most antigenic structural African swine fever virus (ASFV) proteins, has been localized by immuno-electron microscopy in the replication factories of infected cells, mainly associated with membranes and immature virus particles. Attempts to inactivate the p54 gene from ASFV by targeted insertion of beta-galactosidase selection marker was uniformly unsuccessful, suggesting that this gene is essential for virus viability. To demonstrate that, we inserted in the TK (thymidine kinase) locus of the virus a construction containing a second copy of the p54 gene and beta-glucuronidase selection marker under the control of p54 and p73 promoters, respectively. Virus mutant clones expressing a second copy of p54 and beta-glucuronidase were used to achieve deletion mutants of the original copy of the gene. Virus mutants expressing only the second inserted copy of p54 and the two selection markers mentioned above were successfully obtained. Therefore, we have demonstrated that the p54 gene product plays an essential role in virus growth, characterizing for the first time in ASFV an essential virus gene. PMID- 8725113 TI - Analysis of the nucleotide sequence of 23.8 kbp from the left terminus of the genome of variola major virus strain India-1967. AB - Sequencing and computer analysis of the nucleotide sequence of variola major virus strain India-1967 (VAR-IND) DNA segment (23 786 bp) covering the left variable region of the viral genome has been carried out. Twenty-nine potential open reading frames were identified. Structure-function organization of the VAR IND DNA segment was compared with previously reported sequences from analogous genome regions of vaccinia virus strains Copenhagen (VAC-COP) and Western Reserve (VAC-WR). Multiple structural differences between the VAR-IND and genome regions were analysed and both VAC-COP and VAC-WR have been found. Possible molecular factors of virulence, virus host range genes as well as differences revealed in the structure of these genes of VAR and VAC will be discussed. PMID- 8725114 TI - Induction of polyploidy in adenovirus E1-transformed cells by the mitotic inhibitor colcemid. AB - Adenovirus-transformed cells were tested for their ability to synthesize DNA in the presence of cell cycle inhibitory drugs. We show that transformed cells are completely resistant to the mitotic inhibitor colcemid, partly resistant to lovastatin, mimosine, aphidicolin and genistein but not to hydroxyurea or thymidine. When treated with colcemid, AdE1-transformed cells continue to synthesize DNA but do not divide and, therefore, become highly polyploid. This effect is dependent on the presence of both E1A and E1B. PMID- 8725115 TI - The bovine herpesvirus type 1 major tegument protein VP8 expressed in recombinant vaccinia virus does not induce significant immunity in mice. AB - We previously reported the characterization of the gene encoding the bovine herpesvirus type 1 (BHV-1) major tegument protein VP8. With the aim of defining the immunological properties of this protein, we constructed a recombinant vaccinia virus (VV-VP8) in which expression of the VP8 gene was regulated by the P7.5 early/late promoter. Since the sequence of the VP8 gene contained a TTTTTNT motif known to serve as a transcription termination signal of vaccinia virus genes of the early class, a second recombinant (VV-VP8-Mut) in which this signal was modified by site-directed mutagenesis was created. Characterization of the recombinant viruses revealed that truncated VP8 mRNA and protein (69 kDa) were synthesized in VV-VP8 infected cells, whereas cells infected with VV-VP8-Mut produced a protein which was undistinguishable from that of the BHV-1 encoded protein (92-94 kDa). Immunization of BALB/c mice (H-2d) with VV-VP8-Mut induced a low VP8-specific antibody response whereas no specific response was induced in VV VP8 inoculated mice. The low humoral response elicited was similar in C57BL/6 (H 2b) and C3H (H-2k) mice. Furthermore, immunization of mice with VV-VP8-Mut did not induce a BHV-1-specific lymphoproliferation in the three mice strains examined. Our results contrast with a recent study showing that immunization of calves with purified VP8 stimulated both T cell proliferation and antibody production. PMID- 8725116 TI - Purification, characterization and immunogenicity of recombinant varicella-zoster virus glycoprotein gE secreted by Chinese hamster ovary cells. AB - The gene of the varicella-zoster virus (VZV) glycoprotein gE, engineered to code for a truncated molecule lacking the anchor and carboxy-terminal tail domains, was transfected into Chinese hamster ovary (CHO) cells via the pEE14 mammalian expression vector. One recombinant cell line, CHO-gE-2-9, secreted high levels of truncated gE into the culture medium. The product was purified to near homogeneity by a combination of anion exchange, hydrophobic and metal-chelate chromatographies. Purified recombinant gE showed the expected amino-terminal sequence and its glycosylation pattern proved similar to that of the natural product. When injected into mice, using either Freund's or alum as adjuvant, the native truncated gE induced complement-dependent neutralizing antibodies. In contrast, when the molecule was first denatured, it lost immunogenicity with alum. These data show that the recombinant gE, although truncated, could potentially be included in a subunit vaccine against VZV infection. PMID- 8725117 TI - Coding sequences of both genome segments of a European 'very virulent' infectious bursal disease virus. AB - The sequences of segment A (encoding the VP2-VP4-VP3 polyprotein and VP5) and segment B (encoding VP1) of a recent, 'very virulent' (VV) European isolate (UK661) of infectious bursal disease virus (IBDV), a birnavirus, have been determined. There are 26 to 36 amino acid substitutions compared to any other type I IBDV within the segment A polyprotein (of these, 15 are unique) and about 50 substitutions within VP1 (of which 16 are unique). There is more variation compared to classical and antigenic variant viruses, of both virulent and attenuated phenotype, in VP1, VP3 and VP4 than in VP2, even though the latter has previously been identified as the most variable protein between different strains of type I IBDV. In VP3 and VP4, UK661 is the most diverged type I IBDV. Thus the origin of the virus is unclear. It is possible that strong functional constraints have preferentially maintained the primary structure of VP2, though the possibility of recombination cannot be excluded. There are no clear candidate mutations to account for the enhanced virulence of the VV IBDV. Polymerase motifs are well conserved in VP1 but there is an amino acid substitution next to the predicted active-site serine of the viral protease (VP4). In addition, there is a conservative substitution close to the postulated VP2-VP4 cleavage site. It is also now apparent that sequences of IBDV segment B (the segment encoding the RNA polymerase) do not group according to serotype (specified by the capsid proteins encoded on segment A), indicating that segment reassortment has occurred. PMID- 8725118 TI - Restitution of the UL56 gene expression of HSV-1 HFEM led to restoration of virulent phenotype; deletion of the amino acids 217 to 234 of the UL56 protein abrogates the virulent phenotype. AB - Recently it was shown that the avirulent phenotype of HSV-1 strain HFEM is correlated to the lack of DNA sequences of the promoter region of the UL56 gene. In order to investigate the role of the UL56 gene of HSV-1 in the process of viral pathogenicity in more detail, a complete copy of the UL56 gene of the virulent HSV-1 strain 17 was inserted within the DNA sequences of the incomplete UL56 gene of the genome of HSV-1 strain HFEM. The UL56 gene of HSV-1 strain 17 comprises 1428 bp corresponding to the nucleotide positions (NP) 11,5967-117,395 of the genome of HSV-1 strain 17 (SacII-DNA fragment) containing the promoter region and the entire UL56 gene with identical transcription termination signals. This particular DNA fragment was inserted into the corresponding region of the genome of HSV-1 strain HFEM by co-transfection experiments in which the beta galactosidase gene served as reporter gene. Those recombinant viruses with the ability to express the UL56 gene were tested for their pathogenicity in vivo. The results of these experiments indicate that the restoration of the viral UL56 gene expression led to the restitution of the virulent phenotype of HSV-1 strain HFEM. The UL56 protein which has been shown to be a component of the virion possesses several characteristic signatures e.g. a hydrophobic domain at the carboxy terminus between amino acid residues 217 and 234 (VFGVVAIVVVIILVFLWR). In order to investigate the role of this particular signature of the UL56 protein in the process of viral pathogenicity, site-specific mutagenesis was performed for removing the carboxy-terminus of the UL56 protein. The deleted region of the DNA sequences of the UL56 gene between NP 1122-1175 corresponds to NP 116 220-116 373 of the viral genome. The DNA sequences of the UL56 gene of virulent HSV-1 strain 17 and F were replaced by DNA sequences of the truncated UL56 gene by co transfection experiments in which the beta-galactosidase gene served as a reporter gene. Those recombinant viruses with the ability to express the truncated UL56 gene were examined for their pathogenicity in vivo. The analysis revealed that the expression of the truncated UL56 protein (without hydrophobic domain 217-234 aa) was not sufficient for the maintenance of the virulent phenotype of HSV-1 strains. PMID- 8725119 TI - Molecular studies of the acute infection, latency and reactivation of equine herpesvirus-1 (EHV-1) in the mouse model. AB - The murine intranasal (i.n.) infection model was used to study the molecular distribution of equine herpesvirus-1 (EHV-1) during acute infection, latency and following a reactivation stimulus. After inoculation, infectious virus was detected in lungs, nasal turbinates, brains and olfactory bulbs during the acute phase. A nested PCR (nPCR) readily detected virus in these tissues and, in addition, virus was detected in spleens and (in the second round of nPCR) in peripheral blood mononuclear cells (PBMC). A digoxigenin-labelled in situ hybridization probe detected EHV-1 DNA in bronchiolar and vascular endothelium in the lungs and in and around germinal centres in the spleens. One month later, although infectious virus was absent from all tissues, the trigeminal ganglia, olfactory bulb and PBMC remained positive for virus DNA although this was detected only on the second round of nPCR. Furthermore, in situ hybridization, using either DNA or RNA probes, suggested that little or no transcription of virus occurred in neural tissues during the 'latent phase'. Following a reactivation stimulus, infectious virus was not isolated from any tissues, however, EHV-1 DNA was detected on the first round of nPCR in olfactory bulb, trigeminal ganglia and PBMC. This suggested a quantitative increase in EHV-1 DNA occurred following reactivation stimulus. The significance of these results is discussed in relation to the molecular state of EHV-1 in different tissues at various stages of infection and the validity of the murine model for studying latency and reactivation of EHV-1 in the horses. PMID- 8725120 TI - Comparative analysis of the attachment protein gene (H) of dolphin morbillivirus. AB - DMV, dolphin morbillivirus, a paramyxovirus of uncertain origin recently emerged in Mediterranean dolphins. This study presents the complete nucleotide sequence of the hemagglutinin (H) gene including the gene boundaries. The single open reading frame of the DMV H gene encodes a protein of 604 residues which exhibits overall sequence characteristics similar to the H genes of other morbilliviruses. When compared to its closest homologues, measles virus (MV) and rinderpest virus (RPV), DMV has, respectively, 44 and 46% of amino acid residues in identical positions. The primary sequence of the DMV H protein is markedly less conserved than that of the fusion protein. The comparative data at the genomic level correspond with cross-neutralization studies with different morbilliviruses. Retrospective serogical studies dating back to 1983 indicate DMV-like infections in whales of the eastern Atlantic. The presented data support and extend previous studies suggesting that this novel morbillivirus is one of the phylogenetically oldest morbilliviruses known to circulate today. The relationship of DMV and established morbilliviruses to the newly emerged candidate morbillivirus infecting horse and man is discussed. PMID- 8725121 TI - The nucleotide sequence of hydrangea mosaic virus RNA 3 exhibits similarity with the RNA 3 of tobacco streak virus. AB - The complete nucleotide sequence of the RNA 3 of hydrangea mosaic virus (HdMV) was determined. It consists of 2268 nucleotides and contains two open reading frames (ORF). ORF 1 encodes for a putative translation product of 293 amino acids which shared 64.9% identity with the 3a protein of tobacco streak virus (TSV). ORF 2 encodes for a putative translation product of 220 amino acids which shared 54.2% identity with the coat protein of TSV. The relationship between the proteins of HdMV and the corresponding proteins of ilarviruses other than TSV was more distant. No zinc-finger-like motif was found in the coat protein of HdMV but the N-terminus of the protein was rich in basic amino acids. Both terminal, non coding regions of HdMV RNA 3 contained repeated sequences with corresponding homologous fragments in the RNA 3 of TSV. On the basis of the similarities between HdMV and TSV that we detected, we propose that HdMV be included in subgroup 1 of the genus Ilarvirus together with TSV. PMID- 8725122 TI - The adenoviral transcription factor, E1A 13S, trans-activates the human tumor necrosis factor-alpha promoter. AB - The 1311 bp TNF-alpha promoter region fused to a luciferase reporter vector was used in a transient transfection system to study the regulation of TNF-alpha promoter activity by E1A 13S in the U937 macrophage cell line and the MLA 144 T cell line. Co-transfections of the TNF-alpha promoter with an E1A expression vector resulted in a strong trans-activation of the promoter in both cell lines. Sequential truncation of the promoter mapped the E1A responsive region to sequences contained between -120 bp and the transcription start site. Truncation to -95 bp caused a dramatic 87% reduction of E1A activation in MLA 144 cells and further truncation to -36 bp caused a complete loss of E1A activation. In U937 cells, each truncation lowered E1A responsiveness but activity was never completely abolished. Site-directed mutagenesis of putative cis-acting sequences in the TNF-alpha promoter identified the AP-1 site as important for E1A trans activation in the U937 cell line; the AP-2 and CRE sites also appeared to contribute to a lesser degree. In contrast, only the CRE mutation caused a reduction in E1A induced activity in the MLA 144 cell line. Co-transfection of the E1A expression vector with expression vectors for the cellular transcription factors AP-1, AP-2 and CREB indicated that none of these transcription factors showed any co-operativity with E1A. Thus, cis-acting sequences which contribute to E1A trans-activation of the TNF-alpha promoter have been delineated. PMID- 8725123 TI - Translation of the reovirus M1 gene initiates from the first AUG codon in both infected and transfected cells. AB - Reovirus mu 2 protein can be expressed via the mouse phosphoglycerate kinase promoter to low levels in stably transfected L cells. To increase mu 2 expression, the terminal regions of the M1 gene cDNA constructs were modified and the effect on mu 2 expression was analyzed. The M1 gene has a single large open reading frame beginning at nucleotide 14 with another, in frame, AUG codon at nucleotide 161 reported to be used for translation initiation. Unexpectedly, deletions of the M1 5' terminal sequence upstream of the reported translation initiation codon, AUG161, resulted in loss of detection of mu 2 expression. When expression was driven by the stronger T7 promoter in the presence of recombinant vaccinia virus expressing the T7 RNA polymerase, constructs with the M1 5' terminal deletion produced a smaller protein product of approximately 68 kDa, compared to approximately 73 kDa for the protein produced from the full-length M1 containing constructs consistent with the loss of 49 amino acids. The amount of shorter mu 2 product was increased by producing an improved 'Kozak' consensus sequence around the AUG codon at nucleotide 161 or by introducing an internal ribosome entry site at this location. Full-length M1 gene constructs produced a protein of the same size as the authentic mu 2 protein from virus-infected cells. It was further shown that the approximately 73 kDa product was expressed when the M1 gene was in different plasmid backgrounds and even when the M1 gene transcript was preceded by a 1 kb gene. This study demonstrated that translation of the reovirus M1 gene initiates from the first AUG codon in both infected and transfected cells. PMID- 8725125 TI - Communication and cytopathology: Part V: The term "suspicious". PMID- 8725124 TI - The expression of the proteins of equine herpesvirus 1 which share homology with herpes simplex virus 1 glycoproteins H and L. AB - Several expression systems were used in studies aimed at characterizing the equine herpesvirus 1 (EHV-1) glycoprotein H and L homologues of HSV-1 (EHV-1 gH and gL) and the products were compared to the authentic proteins synthesized in virus infected cells. Using an in vitro transcription/translation system two gH species were detected (an unprocessed 89 kDa and a processed 116 kDa product). Three low molecular weight proteins were found in the case of gL (21.8 kDa, 22.9 kDa and 26.9 kDa) and these showed a slight reduction in mobility on the addition of microsomal membranes to the reactions. A gL fusion protein was produced in pGEX-2T, expression being confirmed by Western blotting using a gL-specific antiserum raised against a peptide incorporating the 13 carboxyl terminal amino acids of the protein. A gH specific peptide antiserum precipitated both gH and two smaller proteins from EHV-1 infected cells thought to be two forms of gL. Insect cells infected with gH or gL baculovirus recombinants were used to vaccinate C3H (H-2k) mice. Some protection against EHV-1 infection was conferred to the gH inoculated mice. The results will enable further studies on the importance of the gH and gL interaction in the pathogenesis of EHV-1 to be evaluated and their potential in contributing to a subunit vaccine to be assessed. PMID- 8725127 TI - Cytology of small cell carcinoma of the urinary bladder. AB - To characterize the cytologic spectrum of small cell carcinoma of the urinary bladder, a review of 42 urinary cytology specimens from 13 patients with histologically proven tumors was conducted. Patients ages ranged from 45 to 81 yr (mean 68.9). In four tumors, small cell carcinoma was the sole malignant cellular component: all 11 urinary specimens in these patients harbored cells with features of an undifferentiated small cell carcinoma. In the remaining nine tumors, small cell carcinoma appeared with transitional, squamous cell or adenocarcinoma, but in four, small cell carcinoma was the sole invasive component. Almost one third of urinary specimens in this group lacked a small cell component. Neuroendocrine differentiation was confirmed by immunopathology in eleven cases (neuron specific enolase positive in 11 of 12, synaptophysin in 2/11, chromogranin in 2/13, Leu 7 in 2/7), and by ultrastructural analysis in two. Small cell carcinoma is a cytologically recognizable variant of bladder cancer, but admixture with other malignant components may mask its appearance in urinary specimens. PMID- 8725126 TI - Selective expression of CD44 cell-adhesion molecule in thyroid papillary carcinoma fine-needle aspirates. AB - Recent descriptions of numerous pitfalls in the cytologic diagnosis of thyroid papillary carcinoma on fine-needle aspiration biopsy specimens have prompted studies of new ancillary diagnostic methods. We evaluated the potential use of immunocytochemical staining of thyroid fine-needle aspiration biopsy specimens for CD44, a glycosylated cartilage link protein associated with extracellular matrix adhesion and lymphocyte homing. Fourteen of 16 (88%) classic, surgically confirmed, thyroid aspiration biopsies stained intensely positive for this marker, whereas 0 of 30 (0%) similarly-processed benign aspirates from colloid nodules showed immunoreactivity for CD44 antigen. From this study, we conclude that most papillary carcinomas of the thyroid express the celladhesion molecule CD44, which may be of clinical value in confirming the diagnosis on borderline fine-needle aspiration specimens. Further study of CD44 expression may prove of significant interest in explaining the unusual mode of spread and clinical course of this disease. PMID- 8725128 TI - Epithelial-myoepithelial cell carcinoma of the parotid gland: a case report and comparison of cytologic features with other stromal, epithelial, and myoepithelial cell containing lesions of the salivary glands. AB - The salivary glands are the site of many neoplasms characterized by a mixture of epithelial and myoepithelial cells and stomal fragments. Cytologic distinction of these neoplasms may be very difficult but careful attention to cellular details and the interaction between the cells and stroma can correctly classify these neoplasms as monomorphic or pleomorphic adenomas, adenoid cystic carcinomas, or epithelial-myoepithelial cell carcinomas. PMID- 8725129 TI - Comparison of Cyto-Shuttle and cytocentrifuge as processing methods for nongynecologic cytology specimens. AB - The Cyto-Shuttle (Cancer Diagnostics, Inc., Fairfax, VA) monolayer preparation method was compared to our routine cytocentrifuge method in 129 fluid cytology cases. A single sample from each case was split and prepared by each method. The Cyto-Shuttle preparation was superior to the cytocentrifuge preparation in 51% of cases, equal to it in 38%, and inferior to it in 11%: bronchial wash/lavage (45 cases), 38%, 49%, 13%; body cavity fluid (39 cases), 72%, 15%, 13%; urine (18 cases), 56%, 44%, 0%: peritoneal washing (16 cases), 44%, 44%, 12%; and miscellaneous (11 cases), 36%, 55%, 9%. Cyto-Shuttle preparations were superior due to decreased background and increased number of cells per slide; fixation and morphology were generally equivalent to cytocentrifuge preparations. PMID- 8725130 TI - E-cadherin cell-adhesion molecule expression as a diagnostic adjunct in urothelial cytology. AB - E-cadherin (E-CD) is a cell-adhesion molecule that has been associated with invasion and metastasis in a wide variety of human neoplasms. We have recently shown that, although decreased E-CD expression is associated with increased bladder-wall invasion and higher tumor grade of infiltrating transitional cell carcinomas (TCC), E-CD expression in the exophytic portion of pure papillary and papillary-infiltrating TCC is increased over that of normal transitional cells. To evaluate whether E-CD levels could serve as a diagnostic adjunct in urinary cytology specimens, we stained 40 alcohol-fixed bladder-washing cytospin preparations with an avidin-biotin-peroxidase method using a monoclonal antibody to E-CD (Sigma Chemical Co., St. Louis, MO). E-CD expression level was defined as a high-intensity or low-intensity staining increase over background squamous cell staining for the transitional cells in 21 biopsy-proven transitional cell carcinomas with papillary components, and in 19 benign or reactive control specimens. Twenty-one of 21 TCC (100%) showed an increased E-CD level over background, with 13 low-intensity and 8 highintensity cases. Ten of 19 benign cases (53%) showed increased E-CD staining over background, with 8 low-intensity and 2 highintensity cases. This difference between malignant and benign specimens was statistically significant (chi-square test. P approximately 0.001). We conclude that increased E-CD expression in the papillary components of TCC can be identified in urinary cytology specimens, may reflect the physical and chemical structural makeup of papillary architecture, and warrants further study as a diagnostic adjunct in the interpretation of urine cytology specimens. PMID- 8725131 TI - Automated screening for quality control using PAPNET: a study of 638 negative Pap smears. AB - To determine if the PAPNET screening system can be used for quality control to lower false-negative rates for Pap smears 638 manually screened, "negative" Pap smears were subjected retrospectively to the PAPNET screening system. Twenty-nine of the smears came from 18 patients who subsequently had biopsyproven high-grade squamous intraepithelial lesions (SIL). The remaining 609 negative smears were arbitrarily selected as controls. One hundred twenty-eight (128) of the retrospectively reviewed smears (20%) were selected by PAPNET for microscopic referral because of potential abnormalities. Abnormalities were confirmed on 14 of these smears upon microscopic evaluation. Five of these 14 smears were from smears obtained from four of the 18 women with high-grade SIL. The incidence of manually screened false-negatives detected by PAPNET rescreening was 14/638 or 2.2% for the entire patient population in this study and 5/29 or 17.24% for the targeted patients known to have subsequently developed highgrade lesions. The 2.2% decrease in the false-negative rate in this experiment may be partly artificial as this study group was seeded with false-negative cases at high risk for containing missed abnormalities. The implementation of the PAPNET system for quality control may lower false-negative rates for Pap smears. PMID- 8725132 TI - Fine-needle aspiration of low grade adenosquamous carcinoma of the breast. AB - Low-grade adenosquamous carcinoma is an unusual variant of mammary carcinoma. This malignancy generally presents as a palpable mass without mammographic microcalcifications, and fine-needle aspiration may be the initial technique selected for diagnosis. To our knowledge, the cytologic findings associated with this neoplasm have not been reported. We report a case of low-grade adenosquamous carcinoma of the breast in a 57-yr-old woman, initially studied by fine-needle aspiration cytology and confirmed by excisional biopsy. The aspiration biopsy smears were characterized by low cellularity and small disoriented clusters containing uniform cells of small to medium size. Bipolar cells were not seen in the background. The diagnostic features and differential diagnosis of this unusual neoplasm are reviewed. PMID- 8725133 TI - Fine-needle aspiration biopsy of metaplastic carcinoma of the breast: report of a case with abundant myxoid ground substance. AB - Metaplastic carcinoma (MC) is an uncommon neoplasm of the breast. There are several variants of MC depending on the dominant histologic pattern. The components include over infiltrating ductal carcinoma, extensive squamous differentiation and spindle cell proliferation with or without chondroid or asseous heterologous elements. In FNA smears, only 57% of cases show both ductal carcinoma and metaplastic component. Thus, in almost one half of the cases, the diagnosis is not possible by FNA. Often it is difficult to define the epithelial or sarcomatous character of malignant cells. We describe a case of metaplastic carcinoma of the breast studied by fine-needle aspiration cytology in which myxoid ground substance was the dominant feature in the cytology smears. The rest of the material was composed of scanty isolated atypical cells with large and irregular nuclei. It is important to bear in mind the diagnosis of MC and make a careful search for atypical cells when the cytological smears are mainly composed of myxoid ground substance. PMID- 8725134 TI - Fine-needle aspiration cytodiagnosis of adenoid cystic carcinoma of the breast. AB - Adenoid cystic carcinoma (ACC) of the breast is a rare breast cancer with a good prognosis. Its morphology is identical to its counterpart in the salivary glands. In this brief report, two cases of ACC of the breast are presented in which the diagnosis was established on a fine-needle aspiration cytology (FNAC) and correlated with subsequent examination of cell blocks of aspirate and tissue. The cytodiagnostic criteria for ACC of the breast are discussed. PMID- 8725135 TI - Psammoma bodies in endometriosis: clinical, cytological, and physiopathological implications. AB - This, to the best of our knowledge, is the first report of the cytological observation of psammoma bodies in an endometrioma. Psammoma bodies are laminated, calcified spherites, commonly associated with meningiomas, ovarian serous cystadenocarcinomas, and papillary carcinoma of the thyroid. Many other pathologic processes can produce dystrophic calcification in the form of psammoma bodies. The physiopathological theories of the origin of dystrophic calcification and psammoma bodies will be discussed. The most promising theory implicates matrix vesicles as the nidus for calcification. PMID- 8725136 TI - Biliary tract cytology in specimens obtained by direct cholangiographic procedures: a study of 74 cases. AB - A retrospective review of bile (BL) and biliary tract brushings (Br) obtained by endoscopic retrograde cholangiopancreatography (ERCP) and percutaneous transhepatic cholangiography (PTC) was undertaken to determine the sensitivity and specificity of cytology in the diagnosis of pancreaticobiliary malignancies. A total of 104 cytologic specimens (PTC-BL 15, PTC-Br 13, ERCP-BL 8, ERCP-Br 68) received between 1990 and mid-1994 from 77 patients who had undergone ERCP and/or PTC primarily for biliary stricture were reviewed. Specimens were unsatisfactory/ inadequate in 11 (10.6%), benign in 41 (39.4%), suspicious in 25 (24%), and positive for malignant cells in 27 (26%). Follow-up was available in 74/77 patients; 46 (59.7%) had tissue confirmation while 28 (32.5%) had adequate clinical follow-up based on chart review. Of those with histologic confirmation, there were 32 malignant and 14 benign cases. The overall sensitivity and specificity of PTC- and ERCP-obtained cytologic specimens were 88.9 and 95.7% respectively. There was only one false positive case (ERCP-Br). Overall positive predictive value was 96% negative predictive value 88%, and accuracy 96%. PTC had a significantly lower sensitivity rate (42.8%) and higher rate for unsatisfactory specimens (21%) compared with ERCP-obtained material (100 and 1.9%). Bile obtained by PTC or ERCP appeared less sensitive in detecting malignancies compared with endoscopic brushing using either technique (BL 50% vs. Br 100%). All three false negative cases were PTC-BL specimens. Of the 17 suspicious cases, eight were confirmed histologically as malignant, four were clinically consistent with malignancy, and five showed marked inflammatory atypia on biopsy. Positive predictive value and accuracy rate of a "suspicious cytology" diagnosis were 69 and 80.5%, respectively. Inadequate specimen, poor cellular preservation, and cells obscured by bile all interfere with proper cytologic evaluation. Experience is necessary to appreciate subtle malignant changes in well differentiated carcinomas. Communication between the cytopathologist and the clinician is critical in the accurate interpretation and proper management of the patients. PMID- 8725137 TI - "Arias-Stella reaction"-like changes in endocervical glandular epithelium in cervical smears during pregnancy and postpartum states--a potential diagnostic pitfall. AB - A wide spectrum of histologic changes has been described throughout the female genital tract during pregnancy and in the postpartum period. Of these, the endometrial glandular changes referred to as the Arias-Stella reaction have classically been a diagnostic pitfall in histologic sections. Pregnancy-related changes are also reflected in cytologic material obtained from the cervix and vagina. Both glandular and stromal alterations may be seen. The changes involving endocervical glandular epithelium are often alarming enough to cause diagnostic difficulties, especially when the history of pregnancy is not provided. We report 13 cases where marked glandular changes led to diagnostic misinterpretations. These were characterized by cyto- and karyomegaly, a high nuclear to cytoplasmic ratio, round to oval nuclei with smudgy chromatin imparting a ground glass appearance, frequent intranuclear inclusions, and vacuolated to dense variable cytoplasm. The cytologic diagnoses ranged from "glandular atypia" to "suspicious for adenocarcinoma." Follow-up was available in 11/13 cases. In 9/11 cases, subsequent cervical smears on multiple occasions were negative. Cervical biopsies and/or dilatation and curettage in 4/11 cases did not show significant glandular abnormalities. The glandular changes encountered in cytologic material were similar to those described histologically in the Arias-Stella reaction involving the cervix. This similarity and the fact that these changes disappeared upon termination of the pregnancy favors the presumption that they represent the Arias Stella reaction. Awareness of these changes during pregnancy and postpartum may prevent interpretive errors and unnecessary surgical procedures. PMID- 8725138 TI - Cytologic effects of photodynamic therapy in body fluids. AB - Photodynamic therapy (PDT) has been used in phase I clinical trials at the National Institutes of Health for the treatment of malignancies disseminated within the peritoneal and pleural cavities. Review of records revealed 18 patients who were treated with PDT between April 1988-June 1993. Sixty-five pleural and peritoneal fluids, 22 pre- and 43 post-PDT, were available for evaluation. Mesothelial cell changes seen post-PDT included: increased nuclear-to cytoplasmic ratios in 7/18 (39%), cytomegaly in 9/18 (50%), and multinucleation in 12/18 (67%), with Touton-like giant cells in 3/18 (17%). Additional changes noted post-PDT comprised histiocytic aggregates in 9/18 patients (50%), with granuloma-like clusters in 3/18 (17%), acute and chronic inflammation in 13/18 (72%), and eosinophilia in 8/18 (44%). Residual tumor was present in 7/18 (39%) patients post-PDT. In 2 patients with malignant mesothelioma, benign mesothelial cells with cytologic changes post-PDT were difficult to distinguish from malignant cells. Mesothelial cell changes following PDT, specifically increased nuclear-to-cytoplasmic ratios and cytomegaly, should be recognized to avert false positive diagnoses of tumor. In patients with malignant mesothelioma, and less commonly with adenocarcinoma, benign mesothelial cells with changes secondary to PDT may be difficult to distinguish from tumor cells. PMID- 8725139 TI - Cytology of the thyroid gland: pitfalls in aspiration of the fibrotic nodule. AB - Fibrotic changes can be found in a variety of pathologic processes that affect the thyroid gland, and yet fine-needle aspiration (FNA) correlation of these lesions is not well-known. Cytologic findings are described from three different lesions, all with extensive fibrosis, that occurred in thyroid gland FNA. In each case, fibrotic changes resulted in cytologic changes potentially representing pitfalls in aspiration diagnosis. FNA revealed, solely or predominantly, fibroblasts in 2 patients whose subsequent thyroidectomies disclosed diffusely sclerotic papillary carcinoma and multinodular goiter. In the third case, an initially suspicious FNA was followed by multiple unsuccessful attempts at a second FNA. At thyroidectomy, a follicular adenoma with post-FNA fibrosis and infarction was identified. The differential diagnoses in thyroid FNA of fibrotic nodules can be broad, and this is discussed. Awareness of potential pitfalls may improve diagnostic accuracy. PMID- 8725140 TI - Liquid fixation of endometrial brush cytology ensures a well-preserved, representative cell sample with frequent tissue correlation. AB - Endometrium was obtained by brushing 656 hysterectomy specimens with a MedScand cytobrush (Hollywood, FL) in order to evaluate linking an easy-to-use liquid fixative with brush-sampling of the endometrium, and to determine the fixative's operating characteristics when applied to brush-sampling the endometrium. Liquid fixed brush-samples yielded 4 to 6 slides per case, and any one slide accurately represented the case diagnosis. Tissue fragments were incidentally collected by the brush in 524 cases (79.9%), were least often obtained from senile and weakly proliferative endometria, and were always obtained from high-grade endometrial hyperplasias and carcinomas. With tissue fragments obtained by brush-sampling, the diagnosis matched that of the hysterectomy endometrium. Cytology alone separated benign endometrium from high-grade atypical hyperplasia and carcinoma, showing that clinical decision-making can be based on endometrial brush cytology collected into a liquid fixative. Furthermore, the addition of tissue-fragment histology increased the diagnostic accuracy of the endometrial brushing procedure to 92.5% overall and 100% for high-grade atypical hyperplasias and carcinomas. At this juncture, we advocate endometrial brushing as a sensitive and generally specific case-finding technique; and perhaps as experience teaches us to distinguish low-grade nonatypical hyperplasias from purely physiological changes of the endometrium, it may become accepted as a definitive diagnostic method. The advantages of examining a liquid-fixed suspension are that 1) because these preparations are homogeneous from slide to slide, with any one slide affording the same diagnosis as any other slide from the same case, the number of slides examined may be greatly reduced, and 2) tissue fragments can be used to quality control the cytology diagnosis in the majority of cases, especially in cases of endometrial hyperplasia and carcinoma. PMID- 8725141 TI - Atypical reparative change on cervical/vaginal smears may be associated with dysplasia. AB - The Bethesda System for classifying cervical/vaginal smears has divided reparative changes into two categories: typical and atypical. Although atypical repair may be grouped with atypical squamous cells of unknown significance (ASCUS), this study selected cases specifically diagnosed as atypical reparative changes (ARC), which showed streaming sheets of cells and enlarged nuclei with nucleoli and anisonucleosis. Our goal was to determine the validity of grouping atypical repair with ASCUS. All cases were received by the Medical College of Virginia-Virginia Commonwealth University (MCV-VCU) pathology department in a 6 month period in 1993. Of 196 cases, 55 (28%) were biopsied within a subsequent 3 6-month period. Of these, 27 also carried a diagnosis of ASCUS (9), low-grade squamous intraepithelial lesion (LGSIL) (15), or high-grade squamous intraepithelial lesion (HGSIL) (3). The remaining 28 were presumably biopsied on the basis of clinical concern regarding a diagnosis of ARC. This group, which represents only 14% of the total with the ARC diagnosis, showed a spectrum of histologic changes from squamous metaplasia and chronic cervicitis to HGSIL. Surprisingly, 25% of these patients (7 of 28) showed changes of LGSIL or higher, including 2 (7%) showing HGSIL. This level of higher grade histologic findings justifies separation of atypical from typical repair. Although there are some cases with a significant histologic abnormality found with a cytologic diagnosis of ARC, the percentage of cases is not nearly as high as the 60% range seen associated with the diagnosis of ASCUS. Due to the lack of consensus on the clinical management of ASCUS, we find no justification to separate the diagnosis of ARC from that of ASCUS. PMID- 8725142 TI - Atypical squamous cells of undetermined significance qualified: a follow-up study. AB - The Bethesda System recommends that the diagnosis of "atypical squamous cells of undetermined significance" (ASCUS) be qualified when possible to indicate whether a reactive process, or a squamous intraepithelial lesion (SIL), is favored. In order to evaluate the utility of this recommendation, 308 cervicovaginal smears diagnosed as ASCUS favoring a reactive (FR) or dysplastic (FD) process were identified. Concurrent or follow-up biopsies and cervicovaginal smear results were reviewed and compared to the follow-up of 103 cervicovaginal smears diagnosed as ASCUS without qualification. A follow-up diagnosis of SIL was found in 46.5%, 29.5% and 26.2% of the ASCUS FD, ASCUS FR, and ASCUS unqualified cases, respectively. A repeated ASCUS diagnosis was found in 20.4%, 21.8%, and 18.4% of the ASCUS FD, ASCUS FR, and ASCUS unqualified cases, respectively. Follow-up of each group was negative for dysplasia in 30.0%, 48.7%, and 54.4% of the ASCUS FD, ASCUS FR, and ASCUS unqualified cases, respectively. These results support the use of an ASCUS FD category for the increased detection of patients with dysplasia. The ASCUS FR category was not helpful in our laboratory. Follow-up studies of qualified ASCUS cases in individual practice settings may be useful in guiding patient management. PMID- 8725143 TI - Malpractice protection: communication of diagnostic uncertainty. AB - Malpractice claims against pathologists for misdiagnosis have been sharply rising, especially in the areas of breast fine-needle aspirations (BFNAs) and cervical (Pap) smears. The current state of medical malpractice law is reviewed as it relates to pathologists' anatomic reports. Communication is one of the best medical malpractice prevention tools. This article examines anatomic pathology reports in terms of the merits of communicating diagnostic error rates to the clinician/patient. In the areas of BFNAs and cervical smears, dissemination of diagnostic error rates in the cytology report is recommended. This would help safeguard against malpractice liability being imposed without showing a deviation by the cytopathologist from reasonable practice standards. PMID- 8725144 TI - Intraoperative touch-imprint cytology of germ cell neoplasms. PMID- 8725145 TI - Role of the peroxisome proliferator-activated receptor (PPAR) in mediating the effects of fibrates and fatty acids on gene expression. AB - The three types of peroxisome proliferator-activated receptors (PPAR), termed alpha, delta (or beta), and gamma, belong to the nuclear receptor superfamily. Although peroxisome proliferators, including fibrates and fatty acids, activate the transcriptional activity of these receptors, only prostaglandin J2 derivatives have been identified as natural ligands of the PPAR gamma subtype that also binds thiazolidinedione antidiabetic agents with high affinity. PPARs heterodimerize with retinoic X receptor (RXR) and alter the transcription of target genes after binding to response elements or PPREs, consisting of a direct repeat of the nuclear receptor hexameric DNA recognition motif (PuGGTCA) spaced by 1 nucleotide (DR-1). Upon activation by fatty acids (FAs) and drugs that affect lipid metabolism, PPARs control the expression of genes implicated in intra- and extracellular lipid metabolism, most notably those involved in peroxisomal beta-oxidation. PPARs partially mediate the inductive effects of fibrates and fatty acids on high density lipoprotein (HDL) cholesterol levels by regulating the transcription of the major HDL apolipoproteins, apoA-I and apoA II. The hypotriglyceridemic action of fibrates and certain fatty acids also involves PPAR and is constituted of: 1) increased hydrolysis of plasma triglycerides due to induction of LPL and reduction of apoC-III expression; 2) stimulation of cellular fatty acid uptake and conversion to acyl-CoA derivatives due to increased expression of genes for fatty acid transport protein and acyl CoA synthetase; 3) increased peroxisomal and mitochondrial beta-oxidation; and 4) decreased synthesis of fatty acids and triglycerides and decreased production of very low density lipoprotein (VLDL). Hence, both enhanced catabolism of triglyceride-rich particles and reduced secretion of VLDL particles contribute to the hypolipidemic effect of fibrates and fatty acids. Finally, PPARs appear to be involved in differentiation processes because activation of PPAR gamma 2 triggers adipocyte differentiation and stimulates expression of several genes critical to adipogenesis. It is suggested that PPARs are key messengers responsible for the translation of nutritional and pharmacological stimuli into changes in gene expression and differentiation pathways. PMID- 8725147 TI - Tissue and cellular specific expression of murine lysosomal acid lipase mRNA and protein. AB - Lysosomal acid lipase (LAL) is essential to the intracellular control of cholesterol and triglyceride catabolism via the low density lipoprotein (LDL) delivery of these neutral lipids to the lysosome. Deficiency of LAL in humans leads to Wolman disease and cholesteryl ester storage disease that result, respectively, in the intralysosomal storage of both neutral lipids or only cholesteryl esters. The mouse and human LAL cDNAs were cloned. The deduced amino acid sequences from the mouse and human LAL had high similarity (95%) and identity (75%) including conservation of the active center motifs (G-X-S-X-G) and five potential N-glycosylation consensus sequences. Tissue specific expression of LAL mRNA and protein in mouse tissues was evaluated by in situ hybridization and immunofluorescence staining, respectively. The LAL mRNA was expressed at low levels in most tissues. High level expression was found in hepatocytes and splenic and thymic cells. Very high level expression was observed in cells of the small intestinal villi, the zona fasciculata and reticularis of the adrenal cortex, pancreatic acini, and renal tubular epithelium. Significant levels of expression were detected in epithelial cells of choroid plexus in developing mouse embryo by day 12, in liver and lung by day 14, and in small intestine and kidney by day 16. Similar distribution of LAL protein was observed by immunofluorescence stain. Our results show that the expression of LAL is regulated in a tissue- and cell-specific manner that corresponds to the pathologic involvement in Wolman disease.-Du, H., D. P. Witte, and G. A. Grabowski. Tissue and cellular specific expression of murine lysosomal acid lipase mRNA and protein. PMID- 8725146 TI - Hepatic lipase mediates the uptake of chylomicrons and beta-VLDL into cells via the LDL receptor-related protein (LRP). AB - The uptake of triglyceride-rich lipoproteins has been described as being mediated by apolipoprotein E and lipoprotein lipase (LpL). Proteoglycans, the LDL receptor, and the LDL receptor-related protein (LRP) are the cellular acceptors. In addition to LpL, hepatic lipase (HL) has been shown to bind to LRP. In this study, the role of HL in lipoprotein uptake was investigated. Human chylomicrons and rabbit beta-VLDL were used as ligands for human hepatoma cells, primary human hepalocytes, normal and proteoglycan-deficient Chinese hamster ovary (CHO) cells, and normal and LDL receptor-deficient human fibroblasts. We show that HL induces stimulation of the uptake of chylomicrons and beta-VLDL into the different cell lines. HL is known to bind to heparan sulfate, and experiments on normal and proteoglycan-deficient CHO cells showed that cell surface proteoglycans are essential for HL-mediated uptake of lipoproteins. To exclude LDL receptor mediated uptake. we performed experiments on LDL receptor-deficient fibroblasts that demonstrated that the LDL receptor was not important for the HL-mediated uptake of lipoproteins. Crosslinking experiments confirmed the binding of HL to LRP on the cell surface. To identify the region of HL involved in the interaction with LRP, we used a C-terminal fragment of LpL, known to inhibit LpL-mediated uptake. HL-mediated lipoprotein uptake was suppressed by this fragment. Our experiments indicate that HL, like LpL, can mediate the uptake of lipoproteins into cells, most probably via a C-terminal binding site. The uptake, initiated by proteoglycan binding, is mediated by LRP. PMID- 8725148 TI - Effect of fatty acids and the aqueous diffusion barrier on the uptake and transport of polychlorinated biphenyls in Caco-2 cells. AB - Polychlorinated biphenyls (PCBs) dissolved in dietary fat are absorbed in the gastrointestinal tract by the enterocytes in combination with the fatty acids proceeding from the lipid hydrolysis in the gut lumen. The effect of fatty acid absorption on the uptake and transport of 14 PCBs in enterocytes was studied using monolayers of the human intestinal Caco-2 cell line as a model system. The diffusive resistance of the unstirred water layer and the facilitating role of mixed bile salt micelles on the PCB uptake were examined by varying the thickness of the unstirred water layer adjacent to the apical membrane. In additional experiments, the polarity of the PCB uptake and transport in Caco-2 cells was determined. The solubility of PCBs in the mixed bile salt-fatty acid micelles was 2.7 to 4.8-fold higher than the solubility in plain bile salt micelles. Both the uptake and transport of PCBs in Caco-2 cells were significantly higher (up to 10 fold) when the PCBs were presented mixed with fatty acids. Reducing the thickness of the unstirred water layer resulted in an increased uptake of PCBs. The PCB uptake in Caco-2 cells exceeded the uptake as expected from monomer diffusion only, indicating that bile salt micelles facilitate the PCB transport over the unstirred water layer. Concentrations of dichlorobiphenyls accumulating in the basolateral medium stayed unexpectedly low, suggesting that Caco-2 cells might possess the capability of metabolizing lower chlorinated biphenyls. Uptake of PCBs into the Caco-2 cells was not significantly different whether the PCBs were presented at the apical side or at the basolateral side. However, transport of PCBs over the cell monolayer was significantly higher when the PCBs were presented at the apical side as compared to the basolateral side, suggesting that the unidirectional transport of lipids and lipoproteins affected the PCB transport as well. Our studies indicate that monolayers of the Caco-2 cell line offer a useful model system for studying the intestinal uptake and transport processes of hydrophobic xenobiotics such as polychlorinated biphenyls. PMID- 8725149 TI - Effects of acute inflammation on plasma retinol, retinol-binding protein, and its mRNA in the liver and kidneys of vitamin A-sufficient rats. AB - The acute inflammatory response to tissue injury and infection is associated with low concentrations of plasma retinol and its specific transport proteins, retinol binding protein (RBP) and transthyretin (TTR). To examine the kinetics and mechanism of hyporetinemia, we have induced acute inflammation with lipopolysaccharide (LPS, from Pseudomonas aeruginosa) in rats with adequate stores of vitamin A. Twenty-four h after treatment with LPS (50 micrograms i.p. per 100 g body weight) or saline and food withdrawal, plasma retinol equalled 0.72 +/- 0.06 mumol/L (mean +/- SEM) in five LPS-treated rats versus 1.35 +/- 0.1 mumol/L in five saline-treated rats (P < 0.01). Plasma, liver, and kidney RBP and TTR concentrations were also significantly reduced, but liver and kidney retinol concentrations did not differ between treatment groups. The relative abundance of RBP mRNA in liver (LPS treatment compared to saline treatment) was reduced as early as 12 h (0.44 +/- 0.15, n = 4 pairs, P < 0.02), and continued to be reduced at 24 h (0.57 +/- 0.12, n = 5 pairs, P < 0.02). In the kidney this ratio did not change significantly due to LPS treatment. The relative abundance of cellular retinol-binding protein (CRBP) mRNA in liver and kidney also was not affected by LPS treatment. We infer from these data that inflammation-induced hyporetinemia results from a reduction in the hepatic synthesis of RBP and secretion of the retinol-RBP complex. Moreover, the results imply that plasma retinol concentration is a poor indicator of vitamin A status during inflammation. PMID- 8725150 TI - ApoE enhances lipid uptake by macrophages in lipoprotein lipase deficiency during pregnancy. AB - A woman with primary lipoprotein lipase (LPL) deficiency developed marked hypertriglyceridemia, pancreatitis, eruptive xanthomas, and unusual palmar xanthomas during pregnancy. Hypotheses to account for the palmar xanthomas were that oxidative modification of triglyceride (TG)-rich lipoproteins occurred due to increased plasma residence time, or that their apolipoprotein E (apoE) content was abnormally elevated. Indices of oxidation of her TG-rich lipoproteins did not support the hypothesis that oxidative changes were a causative factor for her xanthomata. However, degradation of her TG-rich lipoproteins by macrophages was markedly increased (1844 ng/mg protein) during pregnancy as compared to hypertriglyceridemic (with normal LPL) and normotriglyceridemic controls (352 and 126 ng/mg protein, respectively). Post pregnancy the degradation of the subject's TG-rich lipoproteins fell to 289 ng/mg protein. Compositional analysis showed significant enrichment of the particles with apoE (0.97 mass ratio of apoE:apoB during pregnancy, in contrast to 0.38 for normolipidemic controls), and was correlated with the rate of degradation of the TG-rich lipoproteins. Thus, the increased uptake of the TG-rich lipoproteins by macrophages appears to be the result of an unusual enrichment of these lipoproteins with apoE. PMID- 8725151 TI - Activation of LDL receptor gene expression in HepG2 cells by hepatocyte growth factor. AB - The effect of recombinant human hepatocyte growth factor (HGF) on low density lipoprotein (LDL) receptor gene expression was studied in the human hepatoma cell line HepG2. HepG2 cells were incubated with serum-free media in the presence and absence of HGF for various times and 125I-labeled LDL specific binding at 4 degrees C, uptake at 37 degrees C, and the levels of LDL receptor mRNA were measured. Incubation with HGF produced time- and concentration-dependent increases in 125I-labeled LDL binding (2-fold), uptake (2.5-fold), and LDL receptor mRNA (6-fold). HGF increased the rate of LDL receptor gene transcription 4- to 5-fold relative to that of several "house-keeping" genes as measured by nuclear run-on transcription. The half-life of LDL receptor mRNA, measured with actinomycin D, was not increased in HGF-treated cells. The stimulation of LDL receptor expression occurred independently of changes in cellular cholesterol or DNA biosynthesis or total cell protein. HepG2 cells were transiently transfected with plasmids bearing either three copies of repeats 2 and 3 (pLDLR(23)3LUC) or one copy of the LDL receptor promoter from -556 to +53 (pLDLR600LUC) linked to firefly luciferase. Incubation of pLDLR(23)3LUC, or pLDLR600LUC-transfected cells with HGF for 4 or 24 h at 37 degrees C produced a concentration-dependent increase in luciferase activity. A maximal stimulation of 3 to 6-fold was achieved for each construct at an HGF concentration of 100 ng/ml. In contrast, HGF had little or no effect on reporter activity in HepG2 cells transfected with a luciferase reporter plasmid bearing the HMG-CoA reductase promoter extending from -325 to +22. Thus, when compared to the native LDL receptor promoter, multiple copies of repeats 2 and 3 of the LDL receptor promoter can fully support activation of the luciferase reporter gene by HGF, demonstrating that the effect of HGF is mediated through the SRE-1. The lack of HGF effects mediated through the HMG-CoA reductase sterol regulatory element suggests, however, that sterol depletion may not be responsible for the induction of the LDL receptor promoter by growth factors. The signalling pathways or effectors responsible for activation of the LDL receptor and HMG-CoA reductase genes thus differ in their response to HGF. These data suggest that the level of SREBP's reaching the nucleus may be determined by as yet unidentified second messengers as well as by sterols. PMID- 8725152 TI - Phase behavior of isolated skin lipids. AB - Ceramides were isolated from the pig stratum corneum (SC) and mixed in varying molar ratios with either cholesterol or with cholesterol and free fatty acids. The phase behavior of the mixtures was studied by small-(SAXD) and wide-angle (WAXD) X-ray diffraction. Ceramides alone did not exhibit a long range ordering. Upon addition of cholesterol to ceramides, lamellar phases were formed and a hexagonal lateral packing was detected similar to that seen in intact SC. At a cholesterol/ceramide molar ratio of 0.1, only one reflection at 5.9 nm was observed. At a cholesterol/ceramide molar ratio of 0.2, three reflections corresponding to 12.3, 5.56, and 4.26 nm appeared. The reflections were based on two phases. Increasing the cholesterol/ceramide ratio to 0.4, the peak positions were slightly shifted. The diffraction pattern revealed the presence of two lamellar phases with periodicities of 12.2 and 5.2 nm, respectively. The positions of the peaks remained unchanged when the cholesterol/ceramide ratio was increased up to 1.0. At a cholesterol/ceramide molar ratio of 2.0, the intensity of various peaks based on the 12.2 nm phase decreased in intensity. The phase behavior of the cholesterol/ceramide mixtures in a ratio between 0.4 and 1.0 was very similar to that found in intact pig SC in which two lamellar phases with periodicities of 6.0 and 13.2 nm are present. Our data further indicate that the formation of the 5.2 nm lamellar phase requires a higher cholesterol content than the formation of the 12.2 nm lamellar phase. Furthermore, when the relative amount of cholesterol is very high, the 5.2 nm phase is the most pronounced one. Addition of free fatty acids increased the solubility of cholesterol, indicating the role free fatty acids may play for the skin barrier function. The phase behavior of cholesterol/ceramide/fatty acid mixtures was found to be dependent on the chain length of fatty acids used. Namely, addition of short-chain free fatty acids (C14-C18) did not change the periodicity of the 12.2 and 5.2 nm phases, but induced the formation of an additional 4.2 nm phase. In the presence of long chain free fatty acids (C16-C26), the periodicity of the lamellar phases was slightly increased (to 13.0 and 5.3 nm, respectively) but no additional 4.2 nm phase was formed. These results indicate that the lipid phase behavior of the cholesterol/ceramide/free fatty acid mixtures closely mimics that of the intact stratum corneum only in the presence of long-chain free fatty acids. PMID- 8725153 TI - Minimally oxidized LDL is a potent inhibitor of lecithin:cholesterol acyltransferase activity. AB - The oxidation of low density lipoproteins (LDL) has been implicated in the development of atherosclerosis. As a variety of highly reactive lipid peroxidation products can transfer from oxidized LDL to HDL, we evaluated the potential deleterious effects of LDL oxidation on HDL-cholesterol metabolism. To address this issue, we exposed the HDL-containing d > 1.063 g/ml fraction of human plasma to copperoxidized LDL and assessed lecithin:cholesterol acyltransferase (LCAT) activity and apolipoproteinA-I (apoA-I) structure. To determine whether LCAT was directly affected by oxidized LDL, independent of crosslinking of apoA-I, we used an exogenous, [14C]cholesterol-labeled proteoliposome substrate to measure plasma LCAT activity. We observed an inhibition of LCAT activity where copper-oxidized LDL possessing only 2.3 +/- 0.1 and 7.3 +/- 1.4 TBARS produced 24 +/- 3% and 47 +/- 10% reductions in [14C]cholesterol esterification by 1 h, respectively. Copper-oxidized LDL that had been passed through a GF-5 desalting column, while retaining only one-third of its original TBARS, possessed nearly all of its LCAT inhibitory capacity suggesting that the LCAT inhibitory factor(s) was a lipophilic oxidation product. Analysis of polarlipids isolated from copper-oxidized LDL indicated that phospholipid and sterol fractions effectively inhibited LCAT. Copper-oxidized LDL, with as little as 6.3 TBARS, also produced intermolecular crosslinking of apoA-I molecules. Taken together, these data suggest that products of LDL oxidation may adversely affect HDL-cholesterol metabolism by two separate mechanisms: 1) a direct inhibitory effect on LCAT activity and 2) through crosslinking of apoA-I. If occurring in vivo, minimally oxidized LDL may impair cholesteryl ester formation on HDL thereby limiting the ability of HDL to function efficiently in the putative antiatherogenic reverse cholesterol transport pathway. PMID- 8725154 TI - Chylomicron metabolism in rats: lipolysis, recirculation of triglyceride-derived fatty acids in plasma FFA, and fate of core lipids as analyzed by compartmental modelling. AB - Chylomicrons labeled in vivo with [14C]oleic acid (primarily in triglycerides (TG), providing a tracer for lipolysis) and [3H]retinol (primarily in ester form, providing a tracer for the corelipids) were injected into rats. Disappearance of the two labels from plasma and appearance of label in plasma free fatty acids (FFA) were analyzed by compartmental modelling. Both core and TG label distributed into an apparent volume 10-15% larger than the blood volume. Part of this probably represents margination to endothelial-binding-lipolysis sites. An open two-compartmental model for plasma FFA was derived from experiments where unesterified oleic acid complexed to albumin was injected. Applying this model revealed that most of the oleic acid from chylomicron triglycerides mixes with the FFA. The disappearance of chylomicron core label required a model in which the label transfers into a second compartment before it leaves the blood. The rate constant for the transformation was high and predicted that, on average, chylomicron spent less than 2 min in the first compartment. The rate out from the second compartment predicted that about 60% of the core label left blood while, on average, chylomicron retained more than half of its triglyceride molecules, i.e., after rather limited lipolysis. The mechanism by which the core label leaves blood is not clear. Modelling showed that under the assumption that the process is shared by chylomicron triglycerides, about half of them go out by this pathway. Comparing fed and fasted rats, the main differences were in the turnover of FFA and in the extent to which chylomicron TG label reappeared in the FFA. This study indicates that a large fraction of the triglycerides in chylomicrons leave plasma together with the core lipids and that most of the fatty acids from chylomicron triglycerides mix into the same metabolic compartments as do plasma free fatty acids. PMID- 8725155 TI - Lipoprotein lipase in highly vascularized structures of the eye. AB - Ocular tissues are highly dependent on lipid turnover and metabolism, which requires an uptake mechanism for fatty acids from lipoproteins. We studied the activity and expression of lipoprotein lipase (LPL), which catalyzes the hydrolysis of plasma triglycerides, in different ocular regions. Human and bovine eyes were dissected and various specialized anatomical areas were assayed for LPL activity, mRNA, and immunoreactivity. Variable levels of LPL activity were detected in all structures in human and bovine eyes. LPL activity was much higher in vascularized structures, such as ciliary body, iris, and retina than in avascular eye structures, such as vitreous body, lens, and cornea. In both human and bovine eyes, ciliary body contained the highest LPL lipolytic activity. LPL mRNA was detected by reverse transcription followed by polymerase chain reaction (RT-PCR) in all regions of human eyes. By RT-PCR analysis it was shown that bovine eyes contained high levels of LPL mRNA in ciliary body and iris, lower levels were found in retina, optic nerve, and lens, whereas no LPL mRNA could be found in bovine cornea. RT-PCR data, obtained in bovine eyes, agree with the results obtained by Northern blot experiments, confirming the high levels of LPL mRNA in iris and ciliary body. Immunofluorescence experiments performed on human eye samples indicated that the LPL protein is mostly distributed on the choroides, the choriocapillaris, and on the vessels of ciliary body, iris, optic nerve, and retina. The present study demonstrates that active LPL protein is synthesized, secreted, and located among microvessels in several specialized regions of the eye, and suggests that LPL could be involved in the uptake of fatty acids by the ocular tissues. PMID- 8725156 TI - 3 alpha, 7 alpha, 12 alpha-trihydroxy-24-nor-5 beta-cholan-23-sulfonate: synthesis and suitability for the study of cholate transport. AB - In order to facilitate the study of transport processes of unconjugated C-24 bile salts, simple syntheses of 3 alpha, 7 alpha, 12 alpha-trihydroxy-24-nor-5 beta cholan-23-sulfonate (norcholansulfonate) and 3 alpha, 7 alpha, 12 alpha trihydroxy-24-nor-5 beta-[7 beta 5H] cholan-23-sulfonate were devised. The hydrophilic-hydrophobic properties of norcholansulfonate, as determined by its chromatographic behavior as well as by its partition between l-octanol and water, are more similar to those of cholyltaurine than to those of cholate. Self association of norcholansulfonate in phosphate buffer, pH 7.4, with an ionic strength of 150 mM begins at a concentration of about 1 mM, comparable to that of cholyltaurine and cholate, as determined by spectral changes in fluorescence emissions of {N-[7-(4-nitrobenzo-2-oxa-1, 3-diazol)]-7b-amino-3a, 12a-dihydroxy 5b-cholan-24 - oyl}-2'-aminoethanesulfonate (7 beta-NBD-NCT). The apparent CMC value obtained from solubilization of the dye Orange OT, 8.5 mM, is comparable to that of cholytaurine. 7.5 mM, and lower than that of cholate, 9.5 mM. Norcholansulfonate is readily taken up by rat liver and completely excreted unmetabolized into bile with about the same secretion maximum (Tm) as cholyltaurine. Biliary excretion of norcholansulfonate is inhibited by cholyltaurine, and, vice versa, norcholansulfonate inhibits cholyltaurine secretion. Concerning metabolism and excretion, norcholansulfonate with the sulfonate group in the position where cholate has the carboxylate group should behave as an appropriate cholate analogue in mediated transport processes. PMID- 8725157 TI - Role of phospholipase A2 enzymes in degradation of dipalmitoylphosphatidylcholine by granular pneumocytes. AB - The role of phospholipase A2 (PLA2) enzymes in the degradation of internalized dipalmitoylphospharidylcoline (DPPC) by rat granular pneumocytes was evaluated with cells in 24 h primary culture on microporous membranes. In cell sonicates and rat lung homogenates, the transition state analogue MJ33 inhibited acidic (pH 4), Ca(2+)-independent PLA2 (aiPLA2) while p-bromophenacylbromide (pBPB) inhibited alkaline (pH 8.5), Ca(2+)-dependent PLA2 and phospholipase C activities. With intact cells, degradation of [3H]methylcholine-labeled DPPC during 2 h incubation was inhibited 48% by MJ33, 20% by pBPB, and 69%by the combination. The inhibitors (20 microM pBPB, 3 mol% MJ33) had no effect on cellular dye exclusion, adherence to membranes, or uptake of DPPC. Arachidonyl trifuoromethylketone, a cytoplasmic PLA2 inhibitor, had no effect on cellular degradation of DPPC. Degradation was depressed approximately 20% by the addition of NH4Cl or methylamine to the medium, suggesting a role for an acidic intracellular compartment in DPPC metabolism. Subcellular fractions prepared by differential centrifugation of rat lung homogenates showed highest specific activity of aiPLA2 in the lamellar body and lysosomal fractions, lower activity in cytosol, and essentially no activity in mitochondria, microsomes, or plasma membranes. The results of this study indicate that aiPLA2 has the major role in the degradation of internalized DPPC by granular pneumocytes and they are compatible with participation of lysosomes/lamellar bodies in DPPC metabolism. PMID- 8725158 TI - Uncoupling of biliary phospholipid and cholesterol secretion in mice with reduced expression of mdr2 P-glycoprotein. AB - Mice in which the gene for mdr2 P-glycoprotein has been disrupted have a severe deficiency in biliary phospholipid and cholesterol secretion. We studied the relation between mdr2 gene expression and biliary lipid secretion with emphasis on the role of bile salt hydrophobicity. Control mice (+/+), and mice with a homozygous (-/-) or heterozygous (+/-) disruption of the mdr2 gene, were infused with taurodeoxycholate (TDC) or tauroursodeoxycholate (TUDC). In mdr2 (-/-) mice, virtually no phospholipids were secreted into bile, irrespective of the type of bile salt infused. In contrast, cholesterol secretion in (-/-) mice increased upon TDC infusion from less than 0.1 to more than 2 nmol/min . 100 g, which was similar to controls under the same conditions. After infusion of TUDC in (-/-) mice. cholesterol secretion also rose (to 1.8 nmol/min . 100 g) but remained much lower than in controls (8 nmol/min x 100 g). In (+/-) mice, cholesterol secretion was equal to (+/+) mice during secretion of endogenous bile salts and during TDC infusion, but was 50% of control levels during maximal TUDC infusion. We conclude that biliary phospholipid secretion completely depends on mdr2 gene expression but cholesterol can, at least partially, be secreted in an mdr2 Pgp-independent mechanism. The extent to which cholesterol is secreted via this mechanism may depend on the hydrophobicity (i.e., cholesterol-solubilizing capacity) of the secreted bile salt. PMID- 8725159 TI - Structural relationships between nascent apoA-I-containing particles that are extracellularly assembled in cell culture. AB - Apolipoprotein A-I (apoA-I) incubated with CHO cells assembles three major nascent lipid complexes with diameters of 7.3, 9, and 11 nm. Previous studies suggested that the smaller nascent particles were precursors for the larger nascent ones. To test this hypothesis, the 7.3, 9, and 11 nm apoA-I-lipid complexes formed by incubating CHO cells with lipid-free apoA-I were isolated and subsequently each subpopulation was re-incubated with cells in the absence of other subpopulations. The physical-chemical characteristics of each subpopulation were examined before and after re-incubation in an effort to understand relationships. if any, between the different nascent complexes. The 7.3, 9, and 11 nm complexes were unique in that each of the particles had pre-alpha mobility on agarose gels: this rapid migration was not altered by re-incubation with cells. Protein crosslinking studies indicated that the 7.3, 9, and 11 nm complexes possessed 2, 3, and 4 apoA-I molecules per complex, respectively; it is unlikely that the size of the particle and number of apoA-I molecules per particle played a role in the increased negative charge of the particles. The present study shows that smaller particles did not give rise to larger ones upon re-incubation with cells. Rather, the 11 and 9 nm particles both generated smaller discs (the 11 nm giving rise primarily to 9 nm discs and the 9 nm complex giving rise to 7.3 nm discs) suggesting that, during incubation with cells, the complexes are destabilized and remodeled into smaller, not larger, complexes. Surprisingly, the 7.3 nm complexes during re-incubation with cells were extremely stable and did not undergo size alteration. When the 7.3 nm particles were incubated with additional small quantities of lipid-free apoA-I (1-2 microgram/ml), larger discoidal complexes were generated suggesting that the formation of larger particles may be driven by the availability of lipid-free apoA-I. PMID- 8725160 TI - Abnormal lipoproteins in the ANIT-treated rat: a transient and reversible animal model of intrahepatic cholestasis. AB - The alpha-naphthylisothiocyanate (ANIT)-treated rat was evaluated as a model for lipoprotein metabolism in cholestatic liver disease. Alterations in lipoprotein composition over a period of 120 h after ANIT treatment (100 mg/kg) were studied. Eighteen hours after treatment, plasma bilirubin and bile acid levels began to rise, together with significant increases in free cholesterol. C-18/16, C-18/18, and C-18/20 phospholipid molecular species. By 48 h, plasma lipid levels were maximal, free cholesterol was 935%, cholesteryl ester 294%, phospholipid 611%, and triacylglycerols 176% of controls, and the cholesteryl ester to free cholesterol ratio began to recover with a modest shift from cholesteryl esters containing C-20 fatty acids to those containing C-18 fatty acids. Lecithin: cholesterol acyltransferase activity was near normal, lipoprotein lipase activity was increased, and hepatic triacylglycerol lipase activity was decreased. Density gradient ultracentrifugation of rat plasma demonstrated a marked shift in lipoprotein density towards the low density lipoprotein range, with the increased lipid being associated with apolipoproteins A-I and E. The presence of large 300 400 A particles and the high surface to core lipid ratio in this density range was consistent with the presence of lipoprotein-X-like vesicles. Apolipoprotein B 48 accumulation was observed in the high density fractions (d15 > 1.063 g/ml) suggesting that these rats have impaired lipoprotein remnant removal. All of these increased levels returned to near normal by 120 h. This study demonstrates that ANIT-treatment induces a transient, fully reversible, non-surgical intrahepatic cholestasis that results in many of the plasma lipoprotein abnormalities associated with human hepatic cholestasis and the bile duct-ligated rat. PMID- 8725161 TI - Probing structure and function of VLDL by synthetic amphipathic helical peptides. AB - Class A amphipathic helical peptides have been shown to mimic many properties of exchangeable apolipoproteins. The three analogs of the class A amphipathic peptides were used to probe the structure and function of human very low density lipoproteins (VLDL): 1) 18 residue peptide possessing a single helical domain (18A) with the sequence Asp-Trp-Leu-Lys-Ala-Phe-Tyr-Asp-Lys-Val-Ala-Glu-Lys-Leu Lys-Glu-Ala-Phe; 2) two domains of 18A separated by a Pro (37pA); and 3) and 18A analog with the end groups protected to increase helicity (Ac-18A-NH2). Upon incubation of the peptides with VLDL at a peptide to VLDL, (protein) ratio of 1:1, the 37pA and Ac-18A-NH2 were able to displace most of apolipoprotein (apo) Cs and E from VLDL without alteration in its lipid composition and morphology while 18A had minimal effect. The extent of displacement was a function of the peptide to VLDL ratio. The rank order of displaceability of apolipoproteins on VLDL was apoE > C-III > C-II. The displacement of apoE and/or Cs from VLDL by peptides variably affected the ability of VLDL to interact with purified bovine milk lipoprotein lipase (LpL) and cultured macrophages. Treatment of VLDL with Ac 18A-NH2 markedly lowered its reactivity to LpL and its ability to induce lipid accumulation in cultured macrophages: however, treatment of VLDL with 37pA or 18A only minimally lowered their abilities. Ac-18A-NH2 treatment of VLDL resulted in the increase of apparent K(m) and a decrease of Vmax for lipoprotein lipase (LpL) catalyzed hydrolysis of VLDL triglycerides. When an artificial triglyceride emulsion was used as a substrate of LpL, 37pA, but not Ac-18A-NH2, activated LpL. The above data indicate that 1) amphipathic helical peptides can alter the metabolic and functional properties of VLDL by dissociating the functionally important exchangeable apolipoproteins from VLDL as well as by acting as a functional element of VLDL after their incorporation; and 2) the class A amphipathic peptides having different lipid-associating properties exert significantly different effect on VLDL function. PMID- 8725162 TI - Effect of experimental nephrosis on hepatic lipoprotein secretion and urinary lipoprotein excretion in rats expressing the human apolipoprotein A-I gene. AB - When human apolipoprotein A-I was expressed in transgenic rats, induction of the nephrotic syndrome resulted in plasma A-I levels exceeding 10 mg/ml. Plasma lipids were no higher than in non-transgenic nephrotic rats. To explain this, the livers from four groups of rats were perfused: wild-type controls (WC), high expressor human apoA-I transgenic controls (TrGC), wild-type nephrotics (WN), and high expressor transgenic nephrotics (TrGN). Compared to the WC group, TrGC rats secreted the same amount of d < 1.063 g/ml lipoproteins but 50% more high density lipoprotein (HDL), with a 5-fold increase in total apoA-I output due to human apoA-I. Compared to the WC group, nephrosis in the WN rats caused a 2-fold increase in both d < 1.063 g/ml lipoproteins and HDL secretion with a 4.6-fold increase in rat apoA-I output. Compared to the TrGC group, nephrosis in the TrGN rats did not increase d < 1.063 g/ml lipoprotein secretion, but caused a 50% increase in HDL secretion and a 6-fold increase in human apoA-I output. The hepatic levels of mRNA for apoB and for HMG-CoA reductase, as well as the degree of apoB mRNA editing, were unchanged. Examination of the perfusate HDL by electron microscopy revealed spherical particles averaging 30 nm in diameter in the WC and WN rats and 17 and 20 nm in the TrGC and TrGN rats. Urinary HDL particles from the TrGN rats did not contain rat apoA-I and averaged 8.2 nm versus 11 nm in the WN rats. We conclude that the size of the nascent HDL, and subsequently of the mature HDL, is determined by the primary structure of apoA-I. In the TrGN rats, the heterogeneous mature HDL has a population of smaller human HDL which is more readily lost in the urine, accounting for the failure of plasma HDL levels to rise above those in TrGC rats. The fact that plasma triglyceride levels in TrGN rats were also not increased may relate to the failure of hepatic apoB secretion to increase, which in turn may have been due to saturation of the protein synthetic capacity by human apoA-I production. PMID- 8725164 TI - Phytanic acid oxidation: normal activation and transport yet defective alpha hydroxylation of phytanic acid in peroxisomes from Refsum disease and rhizomelic chondrodysplasia punctata. AB - In humans the oxidation of phytanic acid is a peroxisomal function. To understand the possible mechanisms for the pathognomic accumulation of phytanic acid in plasma and body fluids of Refsum disease (RD) and rhizomelic chondrodysplasia punctata (RCDP), we investigated activities of various steps (activation, transport, and oxidation) in the metabolism of phytanic acid in peroxisomes isolated from cultured skin fibroblasts from control, RD, and RCDP subjects. Activation of phytanic acid was normal in peroxisomes from both RD and RCDP. Transport of phytanic acid or phytanoyl-CoA in the absence or presence of fatty acid activating cofactors (ATP, MgCl2, and CoASH) into peroxisomes isolated from RD and RCDP skin fibroblasts was also similar to that of peroxisomes from control fibroblasts. Defective oxidation of [(2,3)-3H]- or [1-14C]phytanic acid, or [1 14C]phytanoyl-CoA (substrate for the first step of alpha-oxidation) but normal oxidation of [1-14C] alpha-hydroxyphytanic acid (substrate for the second step of the alpha-oxidation pathway) in peroxisomes from RD clearly demonstrates that excessive accumulation of phytanic acid in plasma and body fluids of RD is due to the deficiency of phytanic acid alpha-hydroxylase in peroxisomes. However, in RCDP peroxisomes, in addition to deficient oxidation of [1-14C]phytanic acid or phytanoyl-CoA or [(2,3)-3H]phytanic acid, the oxidation of [1-14C] alpha hydroxyphytanic acid was also deficient, indicating that in RCDP the activities both of alpha-hydroxylation of phytanic acid and decarboxylation of alpha hydroxyphytanic acid are deficient. These observations indicate that peroxisomal membrane functions (phytanic acid activation and transport) in phytanic acid metabolism are normal in both RD and RCDP. The defect in RD is in the alpha hydroxylation of phytanic acid; whereas in RCDP both alpha-hydroxylation of phytanic acid as well as decarboxylation of alpha-hydroxyphytanic acid are deficient. PMID- 8725163 TI - ATP-dependent phosphatidylcholine translocation in rat liver canalicular plasma membrane vesicles. AB - Phosphatidylcholine (PC) translocation was studied in rat liver canalicular plasma membrane vesicles using a fluorescent PC analogue that permitted the quantitation of asymmetric PC distribution in the outer and inner leaflet of the vesicles. PC translocation to the outer leaflet of the canalicular membrane was stimulated by ATP and an ATP-regenerating system in a time and temperature dependent manner resulting in 200 pmol PC translocated/mg protein per 30 min. A non-hydrolyzable ATP analogue did not support translocation. Translocating activity was observed with PC but not with phosphatidylethanolamine and was specific for inside-out oriented canalicular membrane vesicles. Addition of taurocholate (10 microM), a micelle-forming bile acid, enhanced ATP-dependent PC translocation 1.5 +/- 0.1-fold, whereas addition of taurodehydrocholate (10 microM), a non-micelle-forming bile acid, did not. These results indicate the presence of an ATP-dependent transporter that "flips" phosphatidylcholine from the inner to the outer leaflet of the rat bile canalicular plasma membrane from where it can become associated with bile acids in the canalicular lumen, thereby enhancing ATP-dependent flipping activity. Several lines of evidence suggest that the transporter is Mdr2 P-glycoprotein. PMID- 8725165 TI - Lipid transfer from insect fat body to lipophorin: comparison between a mosquito triacylglycerol-rich lipophorin and a sphinx moth diacylglycerol-rich lipophorin. AB - Two insect lipoproteins, triacylglycerol-rich Aedes aegypti lipophorin and diacylglycerol-rich Manduca sexta lipophorin, were compared in their ability to load neutral lipid from fat body. When fat body of M. sexta was incubated in vitro with [3H]oleic acid, all radiolabeled fatty acids were esterified, predominantly to triacylglycerol. In A. aegypti fat body, however, half of the label remained as free fatty acids. When A. aegypti fat body was radiolabeled with [3H]glycerol, most of the radiolabel was incorporated in triacylglycerol. When either A. aegypti or M. sexta lipophorin was incubated with A. aegypti fat body, labeled with [3H]oleic acid, both lipophorins incorporated mainly radiolabeled free fatty acids, while almost no radiolabeled glycerides were transferred. When the same experiment was performed with A. aegypti fat body, radiolabeled with [3H]glycerol, very little transfer of radiolabeled glycerides was detected. In contrast, when either M. sexta or A. aegypti lipophorin was incubated with M. sexta fat body, both lipophorins incorporated neutral lipids, predominantly diacyglycerol. A. aegypti lipophorin incorporated half the amount of radiolabeled lipid, compared to M. sexta lipophorin. Lipophorins from both species were treated with triacylglycerol lipase of the yeast Candida cylindracea. Although this lipase readily delipidated M. sexta HDLp, it was not able to remove triacylglycerol from A. aegypti HDLp. The data presented suggest that, under the conditions used, lipid transfer from fat body to lipophorin in A. aegypti is not as efficient as in M. sexta. PMID- 8725166 TI - Identification and quantification of diacylglycerols in HDL and accessibility to lipase. AB - We have investigated the presence of diacylglycerols in lipoproteins and especially in HDL. Lipoprotein diacylglycerols are very difficult to isolate and to quantify using classical enzymatic techniques, as they are measured in the presence of triacylglycerols and monoacylglycerols. Using a rapid and very sensitive method of gas-liquid chromatography, developed for neutral lipid analysis on an Ultra 1 Hewlett-Packard fused silica capillary column, diacylglycerols (DG) were identified in HDL and classified into five groups: DG 14-16, DG 16-16, DG 16-18, DG 18-18, and DG 18-20. However, their quantitation was difficult due to only partial resolution of molecular species. HDL lipids were submitted to preparative gas-liquid chromatography and diacylglycerols were then silylated using trimethylsilyl reagents. The trimethylsilyl ethers were analyzed by gas-liquid chromatography on a Restek 50 capillary column and were resolved on the basis of carbon number, degree of unsaturation, and double bond positions. The amount of HDL diacylglycerols was twice that of triacylglycerols. The major molecular species of diacylglycerols consisted of 16:0-18:2n-6, 18:0 18:2n-6, and 16:0-18:1n-9 as the major molecular species (33.4, 22.2, and 16.1 mol % of total diacylglycerols, respectively). Using guinea pig cationic pancreatic lipase in order to test the accessibility of diacylglycerols at the surface of HDL, we measured 59% of diacylglycerol hydrolysis, whereas no triacylglycerol hydrolysis was obtained. In addition, most of diacylglycerols having long chain fatty acids, such as 18-20, were completely hydrolyzed, whereas 18-18 and 16-18 were only partially hydrolyzed (64 and 46% respectively). This reflects a different partition of diacylglycerol molecular species between the particle's surface and the lipid core in HDL. This is the first analysis of diacylglycerol molecular species and their distribution in native lipoprotein particles. PMID- 8725167 TI - Human liposarcoma cell line, SW872, secretes cholesteryl ester transfer protein in response to cholesterol. AB - Cholesteryl ester transfer protein (CETP) mediates the exchange of phospholipids and neutral lipids between the plasma lipoproteins, and plays an important role in high density lipoprotein (HDL) metabolism. While there are reports of low level CETP secretion from cultured cells, the lack of a good model cell line has hampered the detailed study of CETP regulation and secretion. In this study, we have found that the human liposarcoma cell line, SW872, secretes cholesteryl ester transfer protein at levels substantially higher than observed from other cell lines. The secretion of CETP from this adipose-derived cell was up-regulated by 25-OH cholesterol and by low density lipoprotein (LDL) cholesterol in a concentration-dependent manner. Analysis of both full length and exon 9-deleted CETP mRNA demonstrated increases in response to LDL and 25-OH cholesterol, providing evidence for regulation at the message level. Our results suggest that the CETP-producing SW872 cell line may provide a model in which to study the regulation of this important modulator of lipoprotein metabolism. PMID- 8725168 TI - Noninvasive assessment of bone mineral and structure: state of the art. PMID- 8725169 TI - Dietary salt, urinary calcium, and bone loss. PMID- 8725170 TI - A new role for the chondrocyte in fracture repair: endochondral ossification includes direct bone formation by former chondrocytes. AB - We studied the endochondral ossification that occurs during the transition of soft to hard callus during fracture healing in the rabbit. During this process, parts of the cartilaginous soft callus are invaded by capillaries, and new bone is laid down onto the central unresorbed cartilage struts. We found that the chondrocytes within these cartilage struts changed phenotype and became bone forming cells which directly replaced the central cartilage core with bone matrix. We have termed this bone "lacunar" bone to distinguish it from the "vascular" bone laid down by osteoblasts. With time the lacunar bone spread beyond the confines of the lacunae and gradually replaced all the cartilage matrix that was originally present in the early endochondral spicules. The lacunar bone could still be distinguished from the vascular bone as follows: (1) it was woven bone, whereas vascular bone was lamellar bone; (2) it contained acid phosphatase activity, whereas vascular bone did not; and (3) it had strong antigenicity for bone sialoprotein, whereas this noncollagenous protein was undetectable in vascular bone. Eventually the hard callus was resorbed and remodeled, but at an interim period of endochondral ossification the direct replacement of cartilaginous callus by the formation of lacunar bone is a rapid mechanism by which the mechanical strength of fracture callus is increased. PMID- 8725171 TI - Translational regulation of parathyroid hormone gene expression and RNA: protein interactions. AB - The aim of this study was to investigate the mechanism by which translation of parathyroid hormone (PTH) mRNA is regulated with regard to the subcellular distribution of PTH mRNA and RNA:protein interactions. Sucrose density ultracentrifugation of RNA from bovine parathyroid cells indicated that there was no evidence for a pool of nonribosomal PTH mRNA, and the extracellular calcium concentration had no effect on polysome size. UV cross-linking studies revealed two proteins in parathyroid cell cytosol which bound specifically to the 5' untranslated region (UTR) of PTH mRNA with molecular masses of 66 and 68 kD while proteins with apparent molecular masses of 48 and 70 kD bound to the 3'-UTR. In vitro translation assays indicated that parathyroid cell cytosol contains factors that inhibit translation of PTH mRNA. Fractionation of cytosol revealed that this effect was associated with proteins within the molecular mass range 30-90 kD. To determine which sequences in PTH mRNA mediate translational regulation, RNA was synthesized from luciferase gene constructs containing the 5'- and/or 3'-UTR of PTH mRNA, and translated in vitro. Addition of parathyroid cell cytosol reduced the translation of RNA containing the 5'- and 3'-UTR of PTH mRNA by 44 +/- 7% but had no effect on the translation of RNA containing only the luciferase coding region. Translation of RNA containing only the 5'-UTR of PTH mRNA was unchanged; however, cytosol reduced the translation of RNA containing the 3'-UTR by 31 +/- 9%. These data demonstrate a role for RNA:protein interactions in the regulation of PTH synthesis and that translational control is mediated primarily through interactions with the 3'-UTR of PTH mRNA. PMID- 8725172 TI - In vivo demonstration that parathyroid hormone and parathyroid hormone-related protein stimulate expression by osteoblasts of interleukin-6 and leukemia inhibitory factor. AB - We have previously reported that parathyroid hormone (PTH) and PTH related protein (PTHrP) stimulate expression of interleukin-6 (IL-6) and leukemia inhibitory factor (LIF) in osteoblasts in vitro. In the current study, we have developed a model of hormone injection into the subcutaneous space overlying mouse parietal bones to demonstrate that similar processes occur in osteoblasts in vivo. Specifically, PTH and PTHrP rapidly and transiently induce expression of the mRNAs encoding IL-6 and LIF. The effects are dose-dependent, with a maximal stimulation of approximately 50-fold for each cytokine. Although PTH and PTHrP activate both adenyl cyclase and phospholipase C-dependent signal transduction pathways, stimulation of IL-6 and LIF depends on adenyl cyclase since it is not reproduced by PTH(3-34), a partial agonist that only activates phospholipase C. These results confirm our previous in vitro studies and support the hypothesis that IL-6 and/or LIF are physiologically important mediators of at least some of the actions of PTH and PTHrP. PMID- 8725173 TI - Regulation of NFIL-6 and IL-6 expression by basic fibroblast growth factor in osteoblasts. AB - We determined whether basic fibroblast growth factor (bFGF) regulated the expression of IL-6 and NFIL-6 in osteoblasts. In mouse osteoblastic MC3T3-E1 cells, bFGF (10(-8) M) increased NFIL-6 mRNA 2-fold at 30 minutes and 3-fold at 2 h. IL-6 mRNA was increased by bFGF 10(-8) M after 1 h. IL-6 protein was detectable in control cultures but was significantly increased by bFGF (10(-8) M) at 4 h. Immunofluorescence analysis of MC3T3-E1 cells showed primarily cytoplasmic and perinuclear NFIL-6 staining in control cultures while bFGF treated cells showed increased NFIL-6 staining at 2 and 4 h. Western blotting revealed that bFGF increased NFIL-6 protein at 2 h. In calvarial mouse osteoblasts, bFGF 10(-8) M induced IL-6 mRNA as early as 1 h and significantly increased IL-6 protein levels as early as 2 h. In conclusion, bFGF stimulates IL 6 and NFIL-6 mRNA in osteoblasts. The increase in NFIL-6 mRNA was associated with increased NFIL-6 protein. The increase in IL-6 mRNA was also associated with increased IL-6 protein. We propose that activations of NFIL-6 and IL-6 may be important mediators of the effects of bFGF on bone cells. PMID- 8725174 TI - Calbindin-D9k and calbindin-D28k expression in rat mineralized tissues in vivo. AB - Following their terminal differentiation, highly specialized cells, ameloblasts, odontoblasts, and osteoblasts sequentially elaborate mineralized tissues. While the developmental expression pattern of matrix proteins has been studied extensively, less attention has been paid to the molecules involved in calcium handling, such as calcium-binding proteins. This shortcoming, as well as previous conflicting data, led us to conduct studies on calbindin-D9k and calbindin-D28k in rat mandibular bone and incisor based on several methods established on rat ameloblasts in vivo. Radioimmunoassays showed that calbindin-D28k accounts for approximately 0.1% of cytosolic proteins in the ectomesenchymal fraction and 1% in the epithelial fraction of the rat incisor and is 100-fold more concentrated than calbindin-D9k in both tissue types. Western blot analysis confirmed that the anticalbindin-D28k reactive species corresponded to the well characterized renal calbindin-D28k in the ectomesenchyme. In this tissue, calbindin-D28k was ultrastructurally immunolocalized in the odontoblasts. Quantitative immunocytochemistry showed that labeling was distributed throughout their nucleus and cytoplasm. The similar cytoplasmic distribution of both calbindin-D proteins and mRNAs suggests that their expression is regulated at the subcellular level. In particular, immunoreactive calbindin-D28k appeared to be associated with rough endoplasmic reticulum. Calbindin-D9k antisense probe showed negligible labeling in odontoblasts, in parallel with the protein quantities measured (approximately 10 ng/mg of total protein). Finally, in situ hybridization showed transcripts for both calbindins-D in ameloblasts and also in osteoblasts. In summary, the present results support the concept that an elevated expression of these vitamin D dependent calcium-binding proteins may characterize the phenotype of cells directly involved in the elaboration of mineralized tissues, enamel, dentine, and bone. PMID- 8725175 TI - Changes in biological activity of bone cells in ovariectomized rats revealed by in situ hybridization. AB - Twelve-week-old female rats were ovariectomized (OVX) and compared with sham operated control rats at 3, 5, 7, 10, 14, 30, and 60 days postoperation with respect to the expression of type I collagen and osteopontin mRNAs, as well as bone structure and the number of osteoclasts. The trabecular number and separation were significantly decreased and increased, respectively, in the metaphyseal trabecular bones of OVX rat femurs. The number of osteoclasts was significantly increased in the same region of OVX rats at 3 and 5 days postoperation. Type I collagen mRNA was expressed in osteoblasts, and osteopontin mRNA was expressed in some osteoclasts, in mononuclear cells on the bone resorption surface, and in osteocytes near the resorption surface. In the metaphyseal trabecular bone, type I collagen and osteopontin mRNA expression levels in individual cells was initially increased in OVX rats from 7 to 10 days postoperation, and this was sustained for 60 days. The number of osteopontin mRNA expressing osteocytes was also significantly increased at 10 days postoperation, which lasted until 60 days. In the epiphysis, an increase in type I collagen mRNA expression was initially observed in OVX rats at 14 days postoperation, which lasted until 60 days. The number of osteopontin mRNA-expressing osteocytes was virtually identical until 30 days postoperation in the epiphysis. These findings indicated that the biological activities of osteoblasts and osteocytes are stimulated in bones of the OVX rat and that the response for OVX differs between the metaphysis and epiphysis. Furthermore, the number of osteopontin mRNA expressing osteocytes was increased only in bones that tended to be resorbed after OVX. This indicates that some osteocytes were stimulated to express osteopontin mRNA by estrogen deficiency and suggests that these osteopontin mRNA expressing osteocytes may be involved in regulation of bone metabolism. PMID- 8725176 TI - Differential regulation of receptor-stimulated cyclic adenosine monophosphate production by polyvalent cations in MC3T3-E1 osteoblasts. AB - Extracellular cations have paradoxical trophic and toxic effects on osteoblast function. In an effort to explain these divergent actions, we investigated in MC3T3-E1 osteoblasts if polyvalent cations differentially modulate the agonist stimulated cyclic adenosine monophosphate (cAMP) pathway, an important regulator of osteoblastic function. We found that a panel of cations, including gadolinium, aluminum, calcium, and neomycin, inhibited prostaglandin E1 (PGE)-stimulated cAMP accumulation but paradoxically potentiated parathyroid hormone (PTH)-stimulated cAMP production. In contrast, these cations had no effect on forskolin- or cholera toxin-induced increases in cAMP, suggesting actions proximal to adenylate cyclase and possible modulation of receptor interactions with G proteins. Phorbol 12-myristate 13-acetated (PMA) mimicked the effects of cations on PGE1- and PTH stimulated cAMP accumulation in MC3T3-E1 cells, respectively, diminishing and augmenting the responses. Moreover, down-regulation of protein kinase C (PKC) by overnight treatment with PMA prevented gadolinium (Gd3+) from attenuating PGE1- and enhancing PTH-stimulated cAMP production, indicating involvement of PKC dependent pathways. Cations, however, activated signal transduction pathways not coupled to phosphatidylinositol-specific phospholipase C (PI-PLC), since there was no corresponding increase in inositol phosphate formation or intracellular calcium concentrations. In addition, pertussis toxin treatment failed to prevent Gd(3+)-mediated suppression of PGE1-stimulated cAMP, suggesting actions independent of Gm. Thus, polyvalent cations may either stimulate or inhibit hormone-mediated cAMP accumulation in osteoblasts. These differential actions provide a potential explanation for the paradoxical trophic and toxic effects of cations on osteoblast function that occur in vivo under different hormonal conditions. PMID- 8725177 TI - Long-term extracellular matrix metabolism by cultured human osteogenesis imperfecta osteoblasts. AB - Osteopenia due to deficient extracellular matrix synthesis is a hallmark of osteogenesis imperfecta (OI), Previous studies carried out within 72 h of osteoblast subculture, at an early stage of matrix synthesis, indicated that for osteoblasts derived from human OI patients the total amounts of collagen, osteonectin, and three proteoglycans were significantly reduced, while total amounts of thrombospondin, fibronectin, and matrix hyaluronan were elevated compared with age-matched controls. The current study was undertaken to follow OI osteoblast matrix metabolism as that matrix is synthesized, deposited, and matured. Steady-state metabolic radiolabeling was used to follow the metabolism of collagen, hyaluronan, and total proteoglycan by OI and normal osteoblasts for up to 5 weeks. Trabecular osteoblasts from non-OI controls showed an increase in total and matrix-associated collagen synthesis during the first and second week, reaching steady-state levels by week 4. In contrast, cultured OI osteoblasts did not increase either the total (medium + matrix-associated) or matrix-associated collagen during the entire 5-week period. Proteoglycan synthesis exhibited a pattern similar to that for collagen. OI-derived proteoglycans differed from controls in that levels in OI cultures did not reflect the normal time-dependent increase in total proteoglycan and proteoglycan matrix deposition. OI osteoblast hyaluronan synthesis was increased when compared with age-matched controls during 4 weeks of culture. In contrast, the ratios of calcium to phosphorus solublized from control and the OI extracellular matrix were not statistically different. Thus, with respect to the synthesis of collagen, proteoglycans, and hyaluronan, OI osteoblasts fail to parallel controls in depositing and elaborating extracellular matrix during 35 days in culture. PMID- 8725179 TI - Beta-adrenergic receptor kinase-like activity and beta-arrestin are expressed in osteoblastic cells. AB - Biologic responses to peptide calciotropic hormones, such as parathyroid hormone (PTH) and calcitonin, exhibit desensitization. As with most hormones, however, the mechanisms of desensitization are not completely understood. For the beta 2 adrenergic receptor (beta 2AR) system, which is coupled to adenylyl cyclase via the stimulatory guanine nucleotide-binding regulatory (G5) protein, homologous desensitization is mediated in part by a receptor-specific kinase (beta ARK) and a soluble cofactor (beta-arrestin). Recently, this system has been reported to be involved in rapid homologous desensitization of the PTH/parathyroid hormone receptor protein (PTHrP) receptor. We have identified the presence of this system in bone using reverse-transcriptase PCR. Nucleotide sequence of PCR fragments from ROS 17/2.8 cells revealed 100% identity with rat brain beta ARK1 and beta arrestin 1 sequences. Northern analyses with RNA from ROS 17/2.8, UMR 106-H5 cells, and primary cultures of nontransformed neonatal rat calvariae demonstrated two mRNA species of 4 and 2.6 kilobases (kb) for beta ARK and 7.5 kb for beta arrestin, comparable to those found in bovine brain. beta ARK-like activity was demonstrated in cytosolic extracts of the UMR 106-H5 cells by assessing phosphorylation of the retinal photoreceptor, rhodopsin, by the extracts. Phosphorylation was enhanced with light-activated rhodopsin and by bovine brain G beta gamma subunits; heparin inhibited phosphorylation. These findings are characteristic of beta ARK. Expression of beta-arrestin in the UMR 106-H5 cells was confirmed by immunoblot. Thus, osteoblastic cells express proteins, beta ARK, and beta-arrestin, which may regulate desensitization of calciotropic hormone receptors. PMID- 8725178 TI - Development and characterization of a conditionally transformed adult human osteoblastic cell line. AB - Many osteoblastic cell lines are currently in use, but these have limitations either in terms of their relevance to adult human biology and disease or in terms of their suitability for biochemical and molecular analyses. Consequently, we undertook the development of conditionally transformed adult human osteoblastic cell lines. Osteoblasts were obtained from a normal explant cancellous bone chip culture. These cells were infected with adenovirus-ori-SV40 tsA 209, which encodes a temperature-sensitive large T-antigen mutant. Cells immortalized with this virus express a transformed phenotype at the permissive temperature of 34 degrees C but revert to a normal phenotype at the nonpermissive temperature of 40 degrees C. Using this approach, we have isolated several cell clones and describe the characterization of one that was designated HOB-02-C1. Immunocytochemistry revealed that > 95% of the cells express the large T-antigen at both temperatures. These cells exponentially proliferate at 34 degrees C with a doubling time of approximately 2 days but irreversibly stop dividing at 40 degrees C. However, cell volume increases > 2-fold when the cells are maintained for 6 days at the higher temperature. This clone expresses alpha 1 type (I) procollagen mRNA and secretes type I procollagen C-peptide at both temperatures, although the levels were slightly elevated at 40 degrees C. The cell line expresses alkaline phosphatase activity at 34 degrees C, and the basal level of this enzyme increases 2- to 6-fold at 40 degrees C. Alkaline phosphatase activity is induced 4- to 8-fold by 1 alpha,25-dihydroxyvitamin D3 (vitamin D3) at both temperatures, but transforming growth factor-beta 1 (TGF-beta 1) suppresses enzyme expression > 90% at 40 degrees C. Vitamin D3 also induces a 10-fold increase in osteocalcin secretion when the clone is maintained at 34 degrees C, and this induction is enhanced > 8-fold at 40 degrees C. Parathyroid hormone and forskolin stimulate a 4- to 6-fold increase in the production of intracellular cyclic AMP (cAMP) by the cells at 34 degrees C, and this stimulation is enhanced 2- to 4-fold at 40 degrees C. In contrast, prostaglandin E2 stimulates a 7- to 8 fold increase in cAMP only when the cells are maintained at 34 degrees C. This cell line secretes TGF-beta 1 and interleukin-6 (IL-6) at 34 degrees C, but only the basal secretion of IL-6 increases 70% at 40 degrees C. Finally, alizarin red S histochemical staining demonstrates that these cells produce mineralized nodules at both temperatures. In summary, the results of this study indicate that the HOB-02-C1 cells have a mature osteoblastic phenotype. Consequently, this new cell line and others obtained in a similar fashion should be valuable in vitro tools for cellular, biochemical, and molecular studies of adult human osteoblast biology. PMID- 8725180 TI - Vitamin D receptor gene polymorphisms are not related to bone turnover, rate of bone loss, and bone mass in postmenopausal women: the OFELY Study. AB - Vitamin D receptor (VDR) gene polymorphisms have been reported to account for most of the well established genetic influence on bone mineral density (BMD). However, discordant studies have been published and it is still not clear whether VDR genotypes influence bone mass accretion and/or postmenopausal bone loss. In this study, we analyzed VDR gene polymorphisms, i.e., that of BsmI, ApaI, and TaqI restriction enzymes in 268 untreated postmenopausal women 1-26 years postmenopausal. There were 37 BBAA homozygote (absence of BsmI and ApaI restriction sites on both alleles), 55 bbaa homozygote (presence of restriction sites on both alleles), and 176 heterozygotes. At baseline, women between the three genotypes did not differ significantly in age, years since menopause, body mass index (BMI), nor dietary calcium intake. We found no relationship between VDR genotypes and bone turnover assessed by three serum markers of bone formation and three urinary bone resorption markers, nor with BMD measured at the spine, hip, forearm, and whole body by dual-energy X-ray absorptiometry (DXA). Rates of bone loss assessed by repeated DXA measurements over 2 years were highly significant (p = 0.02-0.0001) at all skeletal sites except for the lumbar spine but did not differ between genotypes at any sites either before or after adjustment for potential confounding factors such as years since menopause, BMI, calcium intake, serum 25 hydroxyvitamin D levels, and baseline BMD. When we restricted the analysis to early postmenopausal women, within 10 years of menopause (n = 128), lumbar spine bone loss became significant, but no significant difference between VDR genotypes in the rate of bone loss measured at any site was found. We conclude that VDR genotypes are not predictive of bone turnover, rate of postmenopausal bone loss, and bone mass in either early or late postmenopausal women. In a subgroup of women with a low calcium intake (below 600 mg/day), we also found no significant differences between genotypes in BMD and the rate of bone loss measured at any site, although the sample size (n = 64) may be too small to detect small differences. In conclusion, these data, along with the absence of relationships between VDR gene polymorphisms and peak bone mass that we recently reported, suggest that the determination of VDR genotypes is probably not a useful clinical test for the risk assessment of osteoporosis. PMID- 8725181 TI - A controlled trial of raloxifene (LY139481) HCl: impact on bone turnover and serum lipid profile in healthy postmenopausal women. AB - This randomized, double-blind, placebo-controlled, multicenter, 8-week study evaluated short-term effects of raloxifene on bone turnover, serum lipids, and endometrium in healthy, postmenopausal women. A total of 251 women received either placebo, raloxifene HCl 200 or 600 mg/day, or conjugated estrogens (Premarin, 0.625 mg/day). Bone turnover (serum alkaline phosphatase, serum osteocalcin, urinary pyridinoline cross-links, urinary calcium excretion, urinary hydroxyproline) and serum lipids (total serum cholesterol, high- and low-density lipoprotein cholesterol [HDL-C and LDL-C]) were evaluated at weeks 0, 2, 4, and 8. Endometrial biopsies were performed at weeks 0 and 8. Treatment groups were compared for each parameter for baseline-to-endpoint changes. The estrogen and raloxifene groups experienced similar decreases in serum alkaline phosphatase (range 10-11%), serum osteocalcin (range 21-26%), urinary pyridinoline cross links (range 20-26%), and urinary calcium excretion (range 45-72%). These decreases differed significantly compared with placebo-treated subjects for all markers except serum osteocalcin, the raloxifene HCl 200 mg group. LDL-C decreased significantly in the estrogen and both raloxifene groups (range 5-9%) compared with placebo-treated subjects. HDL-C increased significantly in the estrogen group (16%) but was unchanged in the raloxifene groups. HDL-C:LDL-C ratios increased significantly in the estrogen and raloxifene groups (range 9 29%). Serum cholesterol decreased significantly in both raloxifene groups (range 4-8%) but was unchanged in the estrogen group. Uterine biopsies of raloxifene treated subjects showed no change in the endometrium during this short-term treatment. Biopsies of the estrogen group showed significant endometrial stimulation. The only adverse event possibly related to raloxifene was vasodilatation (hot flashes) which was most common in the raloxifene HCl 600 mg group. Study results indicate that raloxifene may provide beneficial effects to bone and serum lipids in humans without uterine stimulatory effects. PMID- 8725182 TI - Total body calcium and bone mineral content: comparison of dual-energy X-ray absorptiometry with neutron activation analysis. AB - The aim of this study was to compare the dual-energy X-ray absorptiometry (DXA) measurement of bone mass with an independent measure of body calcium obtained by neutron activation analysis (NAA). Total body bone mineral content (BMC) was measured using DXA in 46 subjects in the age range 5-47 years (17 children, 28 young women, and 1 adult male). Total body calcium (TBCa) was measured in the same subjects by in vivo NAA. The correlation between the two measures of bone mass was highly significant (BMC[g] = 3.22 x TBCa[g] - 51.4, r > 0.98, p < 0.0001, SEEBMC = 122.7 g). When BMC was the independent parameter, the SEETBCa was 37.5 g. Bland-Altman analysis indicated a mean difference of 2.8 g with a standard error +/- 4.7 g for TBCaNAA versus TBCaDXA when the BMC values were converted to TBCa. The relative change in bone mass (delta TBCa/delta BMC) for DXA was higher than that reported for dual-photon absorptiometry versus NAA. The findings presented in this study provide translational equations among the DXA and NAA measurements and for the conversion of total body BMC to TBCa in children and young adults. PMID- 8725183 TI - Regional bone mineral density interrelationships in normal and osteoporotic postmenopausal women. AB - We describe a prospective study in which bone mineral density (BMD) was measured in total body and regions, proximal femur, lumbar spine, and forearm in 84 apparently normal postmenopausal women with normal spinal radiographs and in 47 women with 1-10 wedged or compressed vertebrae. There was a history of peripheral fracture in 35 of the 84 controls and 30 of the 47 osteoporotics (p < 0.02) but there was no association between vertebral fracture and wrist fracture. At all sites and regions, the differences in BMD between the "normal"and "osteoporotic" women was highly significant and all but "ribs" and "arms" remained significant after correction for menopausal age. In the whole set, and in both subgroups, the coefficients of correlation between sites and regions were all highly significant (p < 0.001). Nonetheless, some sites discriminated better between the two groups than others. Standardized odds ratios (OR) for vertebral fracture versus no fracture were calculated by logistic regression and expressed as the rise in OR for each standard deviation (SD) fall in bone density. This ratio was greatest (3.4) in "pelvis" and weakest (1.7) in "ribs" but all were statistically significant. Geometric mean regression equations were calculated for all the 78 possible pairs of sites and regions in the 84 normal subjects and the deviations of the osteoporotic women from these normal slopes calculated. In most pairs of sites and regions, the vertebral fracture cases were scattered around the normal group's slope but fell lower down on both axes. The bone deficits in the osteoporotics compared with young normal women ranged from -14% in "head" to -40% in Ward's triangle and the T-scores ranged from -1.9 in "ribs" to -3.9 in the forearm. Sensitivity ranged from 17% in "ribs" to 36.2% in Ward's triangle. Specificity varied between 88 and 94% and the percent correctly classified ranged from 62.6% in "ribs" to 72.5% in Ward's triangle. We conclude that primary postmenopausal osteoporosis affects the entire skeleton but that some sites discriminate better between vertebral fracture and nonfracture cases regardless of whether they represent cortical or trabecular bone. PMID- 8725184 TI - Relationship between body composition and bone mass in women. AB - Increasing body weight is associated both with higher bone mass and with lower rates of bone loss. Whether the effects of body weight are mediated by lean body mass (LBM) or fat body mass (FBM) is, however, uncertain because different studies have used different measures of bone mass and arrived at contradictory conclusions. The parameter actually measured is bone mineral content (BMC). Bone mineral density (BMD), bone mineral apparent density (BMAD), and the BMD/height attempt to "correct" BMC for differences in bone or body size, but these corrections may bias the analysis of the effects of body composition on the skeleton. To resolve this issue, we measured BMC at the total body, lumbar spine, proximal femur, and forearm using dual energy X-ray absorptiometry (DXA) in a population-based sample including 138 premenopausal women (age range 21-54 years, mean 35 years) and 213 postmenopausal women (age range 34-94 years, mean 68 years). BMD, BMAD, and BMD/ height were also calculated for each site. LBM and FBM were determined from the DXA whole body scan. In a multivariate analysis that included age and height, both LBM and FBM predicted total body BMC in pre- and postmenopausal women (p < 0.002 for LBM and FBM in both groups). LBM had a dominant effect on spine and forearm BMC in both groups (p < 0.004) and hip BMC in premenopausal women (p < 0.001), whereas both LBM and FBM predicted hip BMC in postmenopausal women (p < 0.001). However, as BMC was adjusted for bone or body size using BMD, BMAD, or BMD/height, FBM tended to become more important than LBM in the analysis. This was, in part, due to the fact that each of the correction factors in the BMD and BMAD calculations, as well as height, were highly correlated with LBM (r = 0.57 and 0.52 for height versus LBM in pre- and postmenopausal women, respectively [p < 0.001]), and weakly or not at all with FBM (r = 0.08 and 0.11, respectively). Therefore, dividing BMC by these correction factors tended to bias the analysis against potential effects of LBM on bone mass. Thus, the relationship between body composition and bone mass is critically dependent on which bone mass parameter is used in the analysis. Both LBM and FBM have important effects on bone mass, depending on the bone mass parameter used, the skeletal site measured, and menopausal status. PMID- 8725186 TI - Osteopontin deposition in remodeling bone: an osteoblast mediated event. PMID- 8725185 TI - Dimensions and estimated mechanical characteristics of the humerus after long term tennis loading. AB - This study evaluated the effects of long-term unilateral physical activity (tennis) on the playing arm humerus. Total lengths of both humeri, site-specific widths, and the bone mineral contents (BMC) at the proximal, middle, and distal parts of the bones were measured using dual-energy X-ray absorptiometry (DXA). Bone mineral apparent density (BMAD), cortical wall thickness (CWT), cross sectional moment of inertia (CSMI), and section modulus (Z) were approximated from the DXA data for describing the bone's mechanical characteristics more concretely. The study population consisted of 67 healthy, competitive tennis players (17 young men with a mean age [+/- SD] of 25 +/- 5 years, 30 young women with a mean age of 19 +/- 3 years, and 20 older women with a mean age of 43 +/- 5 years) and 57 sedentary controls (16 young men with mean age of 25 +/- 5 years, 25 years, 25 young women with a mean age of 21 +/- 3 years, and 16 older women with mean age of 39 +/- 6 years). All the players had competitive playing histories greater than 4 years. The young male and female players had started their playing careers in childhood (men at the age of 10 +/- 3 years, women 9 +/- 2 years), while the older female players started the training at adulthood (29 +/ 6 years). The playing-to-nonplaying or dominant-to-nondominant arm differences in humeral length ranged from +0.2 to +1.4%, the difference being significant in young male players (+1.4%), young female controls (+1.1%), and older female players (+0.7%). When comparing players' relative side-to-side length differences with those of the controls, no significant differences were found. Significant side-to-side differences in humeral width were observed in all groups except male controls. Compared with the controls, the relative side-to-side width differences were significantly larger at the proximal humerus of the young male players (controls +1.2%, players +3.7%) and the distal humerus of young female players (controls -0.2%, players +1.6%). Compared with the controls, the players' relative side-to-side differences in BMC (range, +7.6 to +25.2%), BMD (+5.8 to +22.5%), BMAD (+5.5 to +20.4%), CWT (+6.9 to +45.2%), CSMI (+7.8 to +26.4%), and Z (+3.0 to +21.7%) were significantly larger in all measured humeral sites except BMAD in the distal humeri of the older female players. These relative side-to side differences were clearly and significantly larger in the young players (+11.7 to +45.2%) than in the older players (+3.0 to +12.4%). In conclusion, long term intensive tennis playing, especially if started in childhood or adolescence, clearly increases the humeral BMC, BMD, and CWT but seems to have only a minor effect on the width of this particular bone. In this respect, there seems to be no sex difference. However, along with the increases in mineral mass and density, the changes in bone width are important in increasing the bending stiffness and strength of the humerus. In older players, the relative side-to-side differences are at the same level or only slightly larger than those in their age-matched controls. This suggests that even intense physical loading of a mature bone is only marginally better in increasing the bone mass, bone density, and CWT of the target bone than the normal daily use of the dominant extremity. PMID- 8725187 TI - A simplified method for the determination of left atrial size and function using cine magnetic resonance imaging. AB - This work aimed at developing a rapid and clinically applicable method for the assessment of left atrial size and function using magnetic resonance imaging (MRI). We studied 17 healthy subjects and 26 cardiac patients. Left atrial cine MRI with 50 ms phases was made in 6-12 contiguous long-axis sections encompassing the entire atrial cavity. A volume-time curve was reconstructed to measure the minimum and maximum volumes as well as the fractional volume change, reservoir function, ejection fraction, and mean filling and emptying rates of the left atrium. The image section with the largest left atrial area was then selected and a comparable area-time curve was reconstructed. The atrial phasic areas and functional indices were determined analogously to the volume-based assessment. The contours of atrial area-time and volume-time curves agreed closely in individual subjects. All area-based left atrial measurements distinguished cardiac patients as a group from healthy persons. The combined specificity of the area-based analyses was 92% and the sensitivity, 65%, in identifying abnormal results in individual patients. The accuracy of the area-based data was best for the atrial minimum size, fractional change, reservoir function, and mean filling rate. The estimated time savings with the simplified method were 5 to 6 h per patient. Left atrial size and function can be studied by reconstructing a phasic atrial area-time curve with cine MRI. Atrial enlargement and abnormalities of filling and reservoir function can be reliably identified, but if data on conduit or stroke function are crucial the three-dimensional MRI technique is still recommended. PMID- 8725189 TI - Approximation of arterial input curve data in MRI estimation of cerebral blood tumor-barrier leakage: comparison between Gd-DTPA and 99mTc-DTPA input curves. AB - Accurate and reproducible MRI measurements of the blood-brain-barrier transport rate constant Ki require measurements of the arterial Gd-DTPA concentration to characterise the arterial input function to the brain. Methods of measuring [Gd] in human plasma are not common practice in clinical departments. In this study we present a fast and easy method of substituting Gd-DTPA with 99mTc-DTPA as the 'tracer' to be measured for the characterisation of the arterial input curve in MRI blood-brain-barrier (BBB) measurements. Gd-DTPA and 99mTc-DTPA was injected intravenously as a bolus into six brain tumor patients during dynamic, fast gradient-echo MRI. Arterial blood samples were collected with an interval of 10(5) during imaging. 99mTc-DTPA and Gd-DTPA were measured in the same blood samples by scintillation counting and Inductively Coupled Plasma Atomic Emission Spectroscopy respectively. The two methods yield the concentrations of both the tracers in mmol/l Gd-DTPA and hence the results are directly comparable. The data results showed no statistically significant difference between the two methods. However, the 99mTc-DTPA method had fewer 'bad points' than the direct Gd-DTPA measurement method. The input curves were in agreement. Conclusively, we found that Gd-DTPA and 99mTc-DTPA were equivalent with regard to distribution and elimination-kinetics during the first 10 min after administration and, therefore, characterisation of arterial input curves may be done using simultaneous injection of 99mTc-DTPA and Gd-DTPA in facilities where means of measuring Gd DTPA is not available. PMID- 8725188 TI - Cerebral arteriovenous malformations: improved nidus demarcation by means of dynamic tagging MR-angiography. AB - Our purpose was to further improve the target volume definition for radiosurgical treatment of cerebral arteriovenous malformations (AVMs) by means of dynamic MRA (dMRA) using a blood bolus tagging sequence. We therefore compare this technique with 3D-TOF-MRA and transfemoral high resolution angiography in plain film technique. Twenty patients with angiographically proven cerebral AVMs were investigated by dMRA, TOF-MRA, and conventional angiography during the MR assisted radiosurgical planning protocol. The patient's head was fixed in an MR compatible stereotactic device. The different angiography techniques were evaluated by consensus of two radiologists. AVMs were characterized by the number and origin of feeding arteries, the maximum diameter of the AVM nidus, and the venous drainage pattern. Dynamic MRA was able to demonstrate the complete AVM characteristics and hemodynamics in 12 out of 20 patients. In three patients with an AVM nidus smaller than 1 cm in diameter the technique could not reliably depict the malformation. Technical problems due to steel screws and pins in the initially used stereotactic frame occurred in five patients. Due to reduced vessel overlap and the lack of disturbances caused by formations with short T1 time, dMRA was superior to TOF-MRA in the detection and the exact localization of the AVM nidus in four patients. We conclude that dMRA is able to demonstrate reliably AVM characteristics and hemodynamics in AVMs with a nidus larger than 1 cm in diameter. Because of the improved demarcation of the AVM nidus, this technique may be a valuable adjunct to radiosurgery planning of cerebral AVMs. PMID- 8725190 TI - The relaxivity of Gd-EOB-DTPA and Gd-DTPA in liver and kidney of the Wistar rat. AB - The NMR relaxivities of Gd-EOB-DTPA and Gd-DTPA were determined in the kidney and liver of intact male Wistar rats immediately following sacrifice and in vitro in solutions and gels, at 1.5 T using a clinical MR scanner, T1 and T2 values of tissue samples were derived from spin-echo image sequences. Tissue gadolinium concentrations were determined by radioassay of Gd153, Gd-EOB-DTPA T1 and T2 relaxivities, R1 and R2 (s-1 mmole-1 kg), were found to be 10.7 +/- 0.5 and 22.5 +/- 3.2 respectively, for liver, 2.4 +/- 0.2 and 12.1 +/- 1.7 for kidney cortex, 2.7 +/- 0.2 and 14.5 +/- 1.9 for kidney outer medulla, 2.0 +/- 0.2 and 11.4 +/- 2.1 for kidney inner medulla. Gd-DTPA R1 and R2 were found to be 4.8 +/- 0.4 and 14.5 +/- 3.7 for liver, 1.2 +/- 0.1 and 7.9 +/- 0.8 for kidney cortex, 1.6 +/- 0.1 and 10.2 +/- 1.4 for kidney outer medulla, 1.3 +/- 0.1 and 10.2 +/- 1.2 for kidney inner medulla. Gd-EOB-DTPA and Gd-DTPA R1 was increased in liver compared to agarose gets at 38 degrees C (4.49 +/- 0.03 and 3.47 +/- 0.06), but reduced in kidney tissues. All R2 were elevated compared to agarose gels at 38 degrees C (5.72 +/- 0.12 and 4.12 +/- 0.03). Elevated R2 and R1 (expressed in terms of the concentration of gadolinium per kg of tissue) can be accounted for in part by the lower water content of tissues compared with gels or solutions increased microviscosity and binding to macromolecules. In addition, susceptibility effects may give rise to further increases in R2. By contrast, the reduced R1 observed in kidney may be the result of compartmentalization of the magnetopharmaceuticals. Statistically improved fits were obtained for T1 recovery curves for liver in the presence of Gd-EOB-DTPA when a dual exponential model was used. Assuming in vitro values for the relaxivities of these artificial contrast agents will lead to inaccuracies when relating observed signal enhancement factors to tissue gadolinium concentration. PMID- 8725191 TI - Binding of manganese and iron tetraphenylporphine sulfonates to albumin is relevant to their contrast properties. AB - The interaction of Fe(III) and Mn(III) complexes of TPPS4 with bovine serum albumin (BSA) was studied by T1 relaxation measurements of water protons and high resolution 1H NMR of the porphyrin moieties. At excess of BSA, both metalloporphyrins bind to BSA as the high spin monomers. The relaxivity of bound MnTPPS4 is significantly higher as compared to the free form in solution. When metalloporphyrins are in excess, they aggregate at the BSA surface, up to two MnTPPS4, and up to 10-15 FeTPPS4 units per BSA globule. Bound aggregates are unable to enhance magnetic relaxation of water protons due to the antiferromagnetic coupling between metal ions in the aggregates. Therefore, the dose-effect dependences for metalloporphyrins in the range of metalloporphyrin/BSA ratio of 0 to 25 at the constant BSA concentration at pH 7.4 are characterized by a local maximum at about 2 for MnTPPS4, and a global maximum at about 3 for FeTPPS4, MnTPPS4 complex is more effective than FeTPPS4 in the whole concentration range. It is suggested that the difference in binding and aggregation properties of metalloporphyrins may be relevant to their relaxation efficiency in vivo, blood transport, and biodistribution. PMID- 8725192 TI - Effects of in utero exposure to 4.7 T MR imaging conditions on fetal growth and testicular development in the mouse. AB - The effects of exposure in utero to high field (4.7 T) magnetic resonance imaging (MRI) conditions on fetal growth, adult growth, and testicular development in the mouse were investigated. Exposures of 8-h duration were made on day 9 and/or day 12 of gestation. Also investigated were effects of a combined treatment regime using continuous wave, unfocused 1 MHz ultrasound on day 9 and MRI on day 12. Treatment groups consisted of 8 to 10 pregnant dams. Fetal data were taken on day 18 of gestation and data from adult animals that were exposed in utero were taken on day 50 postpartum. Significant differences in sensitive biological end points following in utero exposure to complex MRI field conditions were found. Fetal weights of the exposed groups ranged from 11% (day 9 MRI) to 17% (day 9 ultrasound/day 12 MRI) lower (p < or = .05) than those of the cage control and sham-exposed fetuses and crown-rump length of exposed animals was reduced by 8% (p < or = .05). The postpartum death rate was as high as 38% (significant at p < or = .05 level) following MRI exposure on day 9 and day 12 of gestation. Daily sperm production was reduced by as much as 44% (p < or = .05) in adults exposed to MRI on day 12 of gestation. A reduction in testis weight and volume in the adult males was also seen, but the difference was not statistically significant at the p < or = .05 level. No effect was seen on embryonic deaths, sex ration, body weight at day 50, spleen weight, or seminal vesicle weight. The mechanism responsible for these results could involve one or more of the applied fields interacting with a sensitive stage of fetal development. Further studies are planned to isolate the individual field components and to elucidate biochemical and/or hormonal changes that may follow in utero exposure. PMID- 8725193 TI - In situ 19F MRS measurement of RIF-1 tumor blood volume: corroboration by radioisotope-labeled [125I]-albumin and correlation to tumor size. AB - Tumor blood volume (TBV) is an important factor in the metabolism of a tumor and in its response to therapy. Until recently, the only methods to determine TBV were highly invasive and many involved radioisotopes. In this study, a perfluorocarbon (PFC) emulsion, Oxypherol, was monitored by 19F magnetic resonance spectroscopy (MRS). TBVs as determined by 19F MRS of in situ and excised radiation-induced fibrosarcoma (RIF-1) tumors (n = 9), were strongly correlated with the TBV measured by a radioisotope labeled albumin method (slopes of 1.1 and 0.8 with R = 0.86 and 0.91, respectively, by linear regression). In general, the TBV as calculated from the in situ MRS measurements (n = 24) decreased from 28 to 5 ml/100 g tumor mass for tumors ranging in mass from 0.15 to 2 g. However, there was an indication of an initial increase of TBV in tumors smaller than 0.5 g. PMID- 8725195 TI - Quantitation of phosphorus metabolites in newborn human brain using internal water as reference standard. AB - A new method for noninvasive, in vivo quantitation of cerebral phosphorus (31P) metabolites is described. The technique employs point-resolved spectroscopy (PRESS) to obtain both 31P-metabolite and proton (1H) water spectra: brain water is used as an internal concentration reference. Spin-spin relaxation times (T2s) of cerebral 31P metabolites are much longer than the minimum echo time (TE) usable on a spectrometer equipped with actively shielded gradient coils. With short-TE (approximately 10 ms) 31P PRESS, T2 relaxation is minimal and phase modulation of the nucleotide triphosphate (NTP) multiplets can be accounted for 1H water spectra were acquired using several TEs so that extra- and intracellular water signals could be separated from that due to cerebrospinal fluid. Prior calibration of the 31P and 1H spectrometer channels and an assumed brain-water concentration enabled estimations of metabolite concentrations. Using this method, mean 31P metabolite concentrations in the brains of eight normal infants of gestational plus postnatal age 34 to 39 wk were: phosphomonoester (PME) 5.6 (SD 0.9); inorganic phosphate 1.4 (0.4); mobile phosphodiester 2.3 (0.6); phosphocreatine 2.9 (0.3); nucleotide triphosphate 2.8 (0.6); and total mobile phosphate 21.4 (2.8) mmol/kg wet. PMID- 8725194 TI - Reproducibility of metabolite peak areas in 1H MRS of brain. AB - We studied the reproducibility of metabolite signals (from N-acetyl aspartate [NAA], choline, and creatine) measured with a standard single-voxel proton magnetic resonance spectroscopy technique (PRESS, TE = 135 ms, 8 ml VOI) in vitro and in two groups of normal volunteers. Spectral peak areas were quantified both by integration and by curve-fitting. In the in vitro study, the "between-days" variability (coefficient of variation [CV]) of measurements ranged from 0.9% to 2.3%. In the first group of volunteers (n = 12), single voxel spectroscopic measurements (8 ml VOI, 256 acquisitions [ACQs]) were made from mirror-image parts of the right and left hemispheres on 2 separate days. The "between-days" CV of measurements ranged from 9% to 18% for metabolite areas, and from 10% to 26% for metabolite area ratios. There were no significant differences between quantification method or hemisphere. After checking and optimising the MR scanner performance (in fact, it was virtually optimal), the second group (n = 4) each had six sequential single voxel spectroscopic measurements (each of 64 ACQs) from the right hemisphere (without moving the voxel) on each of 4 separate days. Even when the metabolites were measured from the same place in the same hemisphere sequentially six times in a 20-min period, the "within-run" CVs ranged from 4.4% to 17.2% for metabolite areas and from 9.7% to 17.0% for metabolite area ratios. The between-days CVs for the subjects ranged from 7.7% to 25.8% (metabolite areas) and from 10.1% to 22.6% (metabolite area ratios). The variability is due to a combination of random noise, subject motion, baseline artefacts in the spectra, and uncertainties in repositioning the VOIs. It is likely to represent the best reproducibility possible with 8-ml VOIs in cooperative, healthy volunteers carefully positioned on each occasion in a standard clinical scanner. Changes in metabolite levels in individuals must therefore be of the order of 20 40% before we can be reasonably confident of measuring them. Reproducibility in patients, who may be less cooperative, will probably be no better, and this must be taken into account in the interpretation of MRS studies in patients with brain pathology; for example, stroke, head injury, and tumours. PMID- 8725196 TI - Dynamic NMR Q-space studies of microstructure with the multigrade CPMG sequence. AB - A new approach to q-space studies of microstructure is proposed, which exploits the combined information contained in the water proton transverse relaxation time distribution and the frequency dependence of the apparent water diffusivity in heterogeneous systems. Using an automated two-dimensional multigrade CPMG sequence, both the pulse spacing and the amplitude of the applied field gradient are varied systematically and used to measure the frequency and wave vector dependence of the multiple exponential echo decay constants and amplitudes. Undesirable crossterms in the applied and background field gradients are eliminated by a simple procedure involving a sign reversal in the applied gradient. Nonlinear, local susceptibility-induced field gradients are shown to lead to enhanced, frequency-dependent apparent water diffusivities that are sensitive to the local microstructure. PMID- 8725197 TI - NMR imaging of thermal convection patterns. AB - Two special magnetic resonance imaging techniques were applied to the Rayleigh/Benard problem of thermal convection for the first time. The methods were tested using a water cell with horizontal bottom and top covers kept at different temperatures with a downward gradient. Using Fourier encoding velocity imaging (FEVI) a five-dimensional image data set was recorded referring to two space dimensions of slice-selective images and all three components of the local velocity vector. On this basis, the fields of the velocity components or of the velocity magnitude were evaluated quantitatively and rendered as gray shade images. Furthermore the convection rolls were visualized with the aid of two- or three-dimensional multistripe/multiplane tagging imaging pulse sequences based on two or three DANTE combs for the space directions to be probed. Movies illustrating the fluid motions by convection in all three space dimensions were produced. It is demonstrated that the full spatial information of the convection rolls is accessible with microscopic resolution of typically 100 x 100 x 100 microns3. This resolution is effectively limited by flow displacements in the echo time, which should be well within the voxel dimension. The main perspective of this work is that the combined application of FEVI and multistripe/multiplane tagging imaging permits quantitative examinations of thermal convection for arbitrary boundary conditions and with imposed through-flow apart from the direct visualization of convective flow in the form of movies. PMID- 8725198 TI - A comparison of magnetization prepared 3D gradient-echo (MP-RAGE) sequences for imaging of intracranial lesions. AB - In a pilot study including 64 patients with different types of brain tumors we investigated four types of MP-RAGE sequences. The sequences differ in the length of the recovery period and the data acquisition mode (sequential vs. centric phase-encoding). The sequence with sequential encoding and a short recovery period provided images that reached the quality and reliability of spin-echo images. The other MP-RAGE sequences failed in providing equivalent information. In particular, a considerable number of small lesions identified in spin-echo images were not detected in MP-RAGE images. The impact of the evolving magnetization on the point spread function was analyzed by performing simulation calculations. It was found that lesions with short T1 times are rendered with low spatial resolution when sequence parameters are not set appropriately. The low overall quality of images obtained by sequences applying centric encoding may be explained by eddy current effects as reported in other recently published studies. PMID- 8725199 TI - Antiobesity drug, dexfenfluramine, cleared for marketing. PMID- 8725200 TI - H. pylori treatment recognized by FDA. PMID- 8725201 TI - Immunization. PMID- 8725202 TI - Research on the effects of pharmacist-patient communication in institutions and ambulatory care sites, 1969-1994. AB - Research on the effects of pharmacist-patient communication that appeared in the pharmacy literature between 1969 and 1994 is reviewed. The terms patients education and patient counseling were used in identifying relevant research. Many authors used these terms interchangeably; also, the terms counseling and consultation often were not clearly defined. Studies of pharmacists' history taking in institutional settings and of other communication with patients in ambulatory care settings were examined by decade. The research questions, theories, methods, results, and limitations were analyzed. More than 30 studies examined the effect of pharmacists' communication on patient outcomes, primarily knowledge and medication compliance; generally, the results suggested that pharmacists' communication led to increased knowledge and compliance. A few researchers raised concerns about patients' knowledge as an indicator of effective communication by pharmacists, and in the 1980s a few suggested that better medication compliance could be associated with the time and attention given to patients rather than the informational content of the interaction. Little was reported about the communication theories or models on which the studies were based, and there was little indication in most studies that patients' ideas about their therapy were considered. Often, the numbers of patients and pharmacists were small, and the pharmacists may have had training or motivation exceeding that of the average practitioner. In studies of pharmacists' versus physicians' history-taking, the physicians were not well described; their involvement and their approach may not have been comparable to those of the pharmacists. Before 1990, a few researchers had examined outcomes such as pulmonary function and control of diabetes. In the 1990s, more researchers looked at outcomes such as medication-related problems and use of health care resources. The research indicated that pharmacists can increase patients' knowledge of and compliance with their pharmacotherapy; however, additional research on other outcomes may be warranted. PMID- 8725203 TI - Monitoring theophylline therapy to prevent toxicity. PMID- 8725204 TI - Therapeutic-interchange algorithm for multiple drug classes. PMID- 8725205 TI - Stability of ondansetron hydrochloride and five antineoplastic medications. PMID- 8725206 TI - Stability of metoclopramide hydrochloride in plastic syringes. PMID- 8725207 TI - Ceftazidime degradation rates for predicting stability in a portable infusion pump reservoir. PMID- 8725208 TI - Lessons from medicine and nursing for pharmacist-patient communication. PMID- 8725209 TI - Effect of physician education on omeprazole use at a small public hospital. PMID- 8725210 TI - Savings on nutritional supplements administered to patients with chronic alcoholism. PMID- 8725211 TI - Homeopathy? Absurd. PMID- 8725212 TI - More information on vancomycin clearance by new hemodialysis membranes. PMID- 8725213 TI - A metabolic basis for dominance and recessivity. PMID- 8725215 TI - Genetic divergence and fitness convergence under uniform selection in experimental populations of bacteria. AB - Replicate populations of bacteria were propagated for 1000 generations in the laboratory. The growth substrate was periodically renewed, so that during most generations (cell doublings) it was not limiting. The final clones demonstrated about a 40% fitness increase when competed against their common ancestor. This increase was uniform both among and within populations despite extensive differentiation in correlated traits: cell size, resistance to starvation and dry mass of culture. It is suggested that genetic diversity developed because selection promoted any changes directing cell activity toward a higher maximum growth rate. Evolution of this trait halted at a similar level when some basic constraints on bacterial metabolism were met. The selective values of emerging mutations must have depended on the genetic background. They would be beneficial early in evolution but ineffective near the limit of adaptation. This hypothesis was tested for one mutation that affected both fitness and colony morphology. In some clones it was the first adaptive mutation and provided a third of the total fitness increase, but it was not assimilated by the clones that reached the adaptive ceiling in some other way. Near the limit of adaptation, epistasis levels off the fitnesses of genetically variable clones. PMID- 8725214 TI - A mutation in a new gene, bglJ, activates the bgl operon in Escherichia coli K 12. AB - A new mutation, bglJ4, has been characterized that results in the expression of the silent bgl operon. The bgl operon encodes proteins necessary for the transport and utilization of the aromatic beta-glucosides arbutin and salicin. A variety of mutations activate the operon and result in a Bgl+ phenotype. Activating mutations are located upstream of the bgl promoter and in genes located elsewhere on the chromosome. Mutations outside of the bgl operon occur in the genes encoding DNA gyrase and in the gene encoding the nucleoid associated protein H-NS. The mutation described here, bglJ4, has been mapped to a new locus at min 99 on the Escherichia coli K-12 genetic map. The putative protein encoded by the bglJ gene has homolgy to a family of transcriptional activators. Evidence is presented that increased expression of the bglJ product is needed for activation of the bgl operon. PMID- 8725216 TI - A search for a general phenomenon of adaptive mutability. AB - The most prominent systems for the study of adaptive mutability depend on the specialized activities of genetic elements like bacteriophage Mu and the F plasmid. Searching for general adaptive mutability, we have investigated the behavior of Salmonella typhimurium strains with chromosomal lacZ mutations. We have studied 30 revertible nonsense, missense, frameshift, and insertion alleles. One-third of the mutants produced > or = 10 late revertant colonies (appearing three to seven days after plating on selective medium). For the prolific mutants, the number of late revertants showed rank correlation with the residual beta galactosidase activity; for the same mutants, revertant number showed no correlation with the nonselective reversion rate (from fluctuation tests). Leaky mutants, which grew slowly on selective medium, produced late revertants whereas tight nongrowing mutants generally did not produce late revertants. However, the number of late revertants was not proportional to residual growth. Using total residual growth and the nonselective reversion rate, the expected number of late revertants was calculated. For several leaky mutants, the observed revertant number exceeded the expected number. We suggest that excess late revertants from these mutants arise from general adaptive mutability available to any chromosomal gene. PMID- 8725217 TI - Suppression analysis reveals a functional difference between the serines in positions two and five in the consensus sequence of the C-terminal domain of yeast RNA polymerase II. AB - The largest subunit of RNA polymerase II contains a repetitive C-terminal domain (CTD) consisting of tandem repeats of the consenus sequence Tyr1Ser2Pro3Thr4Ser5Pro6Ser7. Substitution of nonphosphorylatable amino acids at positions two or five of the Saccharomyces cerevisiae CTD is lethal. We developed a selection system for isolating suppressors of this lethal phenotype and cloned a gene, SCA1 (suppressor of CTD alanine), which complements recessive suppressors of lethal multiple-substitution mutations. A partial deletion of SCA1 (sca1 delta ::hisG) suppresses alanine or glutamate substitutions at position two of the consensus CTD sequence, and a lethal CTD truncation mutation, but SCA1 deletion does not suppress alanine or glutamate substitutions at position five. SCA1 is identical to SRB9, a suppressor of a cold-sensitive CTD truncation mutation. Strains carrying dominant SRB mutations have the same suppression properties as a sca1 delta ::hisG strain. These results reveal a functional difference between positions two and five of the consensus CTD heptapeptide repeat. The ability of SCA1 and SRB mutant alleles to suppress CTD truncation mutations suggest that substitutions at position two, but not at position five, cause a defect in RNA polymerase II function similar to that introduced by CTD truncation. PMID- 8725218 TI - A novel Ty1-mediated fragmentation method for native and artificial yeast chromosomes reveals that the mouse steel gene is a hotspot for Ty1 integration. AB - We have developed a powerful new tool for the physical analysis of genomes called Ty1-mediated chromosomal fragmentation and have used the method to map 24 retrotransposon insertions into two different mouse-derived yeast artificial chromosomes (YACs). Expression of a plasmid-encoded GAL1:Ty1 fusion element marked with the retrotransposition indicator gene, ade2AI, resulted in a high fraction of cells that sustained a single Ty1 insertion marked with ADE2. Strains in which Ty1ADE2 inserted into a YAC were identified by cosegregation of the ADE2 gene with the URA3-marked YAC. Ty1ADE2 elements also carried a site for the endonuclease I-DmoI, which we demonstrate is not present anywhere in the yeast genome. Consequently, I-DmoI cleaved a single chromosome or YAC at the unique site of Ty1ADE2 insertion, allowing rapid mapping of integration events. Our analyses showed that the frequency of Ty1ADE2 integration into YACs is equivalent to or higher than that expected based on random insertion. Remarkably, the 50-kb transcription unit of the mouse Steel locus was shown to be a highly significant hotspot for Ty1 integration. The accessibility of mammalian transcription units to Ty1 insertion stands in contrast to that of yeast transcription units. PMID- 8725219 TI - The yeast BSD2-1 mutation influences both the requirement for phosphatidylinositol transfer protein function and derepression of phospholipid biosynthetic gene expression in yeast. AB - The BSD2-1 allele renders Saccharomyces cerevisiae independent of its normally essential requirement for phosphatidylinositol transfer protein (Sec14p) in the stimulation of Golgi secretory function and cell viability. We now report that BSD2-1 yeast mutants also exhibit yet another phenotype, an inositol auxotrophy. We demonstrate that the basis for this Ino- phenotype is the inability of BSD2-1 strains to derepress transcription of INO1, the structural gene for the enzyme that catalyzes the committed step in de novo inositol biosynthesis in yeast. This constitutive repression of INO1 expression is mediated through specific inactivation of Ino2p, a factor required for trans-activation of INO1 transcription, and we show that these transcriptional regulatory defects can be uncoupled from the "bypass Sec14p" phenotype of BSD2-1 strains. Finally, we present evidence that newly synthesized phosphatidylinositol is subject to accelerated turnover in BSD2-1 mutants and that prevention of this accelerated phosphatidyl-inositol turnover in turn negates suppression of Sec14p defects by BSD2-1. We propose that, in BSD2-1 strains, a product(s) generated by phosphatidylinositol turnover coordinately modulates the activities of both the Sec14p/Golgi pathway and the pathway through which transcription of phospholipid biosynthetic genes is derepressed. PMID- 8725220 TI - Los1p, involved in yeast pre-tRNA splicing, positively regulates members of the SOL gene family. AB - To understand the role of Los1p in pre-tRNA splicing, we sought los1 multicopy suppressors. We found SOL1 that suppresses both point and null LOS1 mutations. Since, when fused to the Ga14p DNA-binding domain, Los1p activates transcription, we tested whether Los1p regulates SOL1. We found that las1 mutants have depleted levels of SOL1 mRNA and Sol1p. Thus, LOS1 appears to positively regulate SOL1. SOL1 belongs to a multigene family with at least two additional members, SOL2 and SOL3. Sol proteins have extensive similarity to an unusual group of glucose-6 phosphate dehydrogenases. As the similarities are restricted to areas separate from the catalytic domain, these G6PDs may have more than one function. The SOL family appears to be unessential since cells with a triple disruption of all three SOL genes are viable. SOL gene disruptions negatively affect tRNA-mediated nonsense suppression and the severity increases with the number of mutant SOL genes. However, tRNA levels do not vary with either multicopy SOL genes or with SOL disruptions. Therefore, the Sol proteins affect tRNA expression/ function at steps other than transcription or splicing. We propose that LOS1 regulates gene products involved in tRNA expression/function as well as pre-tRNA splicing. PMID- 8725221 TI - Destabilization of simple repetitive DNA sequences by transcription in yeast. AB - Simple repetitive DNA sequences in the eukaryotic genome frequently alter in length. In wild-type strains, we find that transcription through a repetitive poly GT tract destabilizes the tract four- to ninefold. In mismatch repair deficient yeast strains, simple repeats are very unstable. High levels of transcription in such strains destabilize repetitive tracts an additional two- to threefold. PMID- 8725222 TI - Mutagenesis of the yeast gene PRP8 reveals domains governing the specificity and fidelity of 3' splice site selection. AB - PRP8 encodes a highly conserved U5 snRNP protein required for spliceosome assembly and later steps of pre-mRNA splicing. We recently identified a novel allele, prp8-101, that specifically impairs recognition of the uridine tract that precedes most yeast 3' slice sites. We carried out extensive mutagenesis of the gene and selected for new alleles that confer a phenotype similar to that of prp8 101. The strongest alleles cause changes in one of two amino acids in the C terminal portion of the protein. We also identified a second class of PRP8 mutant that affects the fidelity of 3' splice site utilization. These alleles suppress point mutations in the PyAG motif at the 3' splice site and do not alter uridine tract recognition. The strongest of these alleles map to a region directly upstream of the prp8-101-like mutations. These new PRP8 alleles define two separable functions of Prp8p, required for specificity of 3' splice site selection and fidelity of 3' splice site utilization, respectively. Taken together with other recent biochemical and genetic data, our results suggest that Prp8p plays a functional role at the active site of the spliceosome during the second catalytic step of splicing. PMID- 8725223 TI - Meiotic recombination initiated by a double-strand break in rad50 delta yeast cells otherwise unable to initiate meiotic recombination. AB - Meiotic recombination in Saccharomyces cerevisiae is initiated by double- strand breaks (DSBs). We have developed a system to compare the properties of meiotic DSBs with those created by the site-specific HO endonuclease. HO endonuclease was expressed under the control of the meiotic-specific SPO13 promoter, creating a DSB at a single site on one of yeast's 16 chromosomes. In Rad+ strains the times of appearance of the HO-induced DSBs and of subsequent recombinants are coincident with those induced by normal meiotic DSBs. Physical monitoring of DNA showed that SPO13: : HO induced gene conversions both in Rad+ and in rad50 delta cells that cannot initiate normal meiotic DSBs. We find that the RAD50 gene is important, but not essential, for recombination even after a DSB has been created in a meiotic cell. In rad50 delta cells, some DSBs are not repaired until a broken chromosome has been packaged into a spore and is subsequently germinated. This suggests that a broken chromosome does not signal an arrest of progression through meiosis. The recombination defect in rad50 delta diploids is not, however, meiotic specific, as mitotic rad50 diploids, experiencing an HO-induced DSB, exhibit similar departures from wild-type recombination. PMID- 8725224 TI - Homologous and homeologous intermolecular gene conversion are not differentially affected by mutations in the DNA damage or the mismatch repair genes RAD1, RAD50, RAD51, RAD52, RAD54, PMS1 and MSH2. AB - Mismatch repair (MMR) genes or genes involved in both DNA damage repair and homologous recombination might affect homeologous vs. homologous recombination differentially. Spontaneous mitotic gene conversion between a chromosome and a homologous or homeologous donor sequence (14% diverged) on a single copy plasmid was examined in wild-type Saccharomyces cerevisiae strains and in MMR or DNA damage repair mutants. Homologous recombination in rad51, rad52 and rad54 mutants was considerably reduced, while there was little effect of rad1, rad50, pms1 and msh2 null mutations. DNA divergence resulted in no differential effect on recombination rates in the wild type or the mutants; there was only a five to 10 fold reduction in homeologous relative to homologous recombination regardless of background. Since DNA divergence is known to affect recombination in some systems, we propose that differences in the role of MMR depends on the mode of recombination and/or the level of divergence. Based on analysis of the recombination breakpoints, there is a minimum of three homologous bases required at a recombination junction. A comparison of Rad+ vs. rad52 strains revealed that while all conversion tracts are continuous, elimination of RAD52 leads to the appearance of a novel class of very short conversion tracts. PMID- 8725225 TI - DLH1 is a functional Candida albicans homologue of the meiosis-specific gene DMC1. AB - DMC1/LIM15 homologue 1 (DLH1), a gene related to meiosis-specific genes, has been isolated from Candida albicans, a fungus thought not to undergo meiosis. The deduced protein sequence of DLH1 contains 74% amino acid identity with Dmc1p from Saccharomyces cerevisiae and 63% with Lim15p from the plant Lilium longiflorum, meiosis-specific homologues of Escherichia coli RecA. Candida DLH1 complements a dmc1/dmc1 null mutant in S. cerevisiae. High copy expression of DLH1 restores both sporulation and meiotic recombination to a Saccharomyces dmc1 delta/dmc1 delta strain. Unlike the DMC1 gene, which is transcribed only in meiotic cells, the heterologous Candida DLH1 gene is transcribed in both vegetative and meiotic cells of S. cerevisiae. Transcription of DLH1 is not detected or induced in C. albicans under conditions that induce DMC1 and meiosis in S. cerevisiae. The presence of an intact homologue of a meiosis-specific gene in C. albicans raises the possibility that this organism has a cryptic meiotic pathway. PMID- 8725226 TI - Mitochondrial intronic open reading frames in Podospora: mobility and consecutive exonic sequence variations. AB - The mitochondrial genome of 23 wild-type strains belonging to three different species of the filamentous fungus Podospora was examined. Among the 15 optional sequences identified are two intronic reading frames, nad1-i4-orf1 and cox1-i7 orf2. We show that the presence of these sequences was strictly correlated with tightly clustered nucleotide substitutions in the adjacent exon. This correlation applies to the presence or absence of closely related open reading frames (ORFs), found at the same genetic locations, in all the Pyrenomycete genera examined. The recent gain of these optional ORFs in the evolution of the genus Podospora probably account for such sequence differences. In the homoplasmic progeny from heteroplasmons constructed between Podospora strains differing by the presence of these optional ORFs, nad1-i4-orf1 and cox1-i7-orf2 appeared highly invasive. Sequence comparisons in the nad1-i4 intron of various strains of the Pyrenomycete family led us to propose a scenario of its evolution that includes several events of loss and gain of intronic ORFs. These results strongly reinforce the idea that group 1 intronic ORFs are mobile elements and that their transfer, and concomitant modification of the adjacent exon, could participate in the modular evolution of mitochondrial genomes. PMID- 8725227 TI - Pseudohomothallism and evolution of the mating-type chromosome in Neurospora tetrasperma. AB - Ascospores of Neurospora tetrasperma normally contain nuclei of both mating-type idiomorphs (a and A), resulting in self-fertile heterokaryons (a type of sexual reproduction termed pseudohomothallism). Occasional homokaryotic self-sterile strains (either a or A) behave as heterothallics and, in principle, provide N. tetrasperma with a means for facultative outcrossing. This study was conceived as an investigation of the population biology of N. tetrasperma to assess levels of intrastrain heterokaryosis (heterozygosity). The unexpected result was that the mating-type chromosome and autosomes exhibited very different patterns of evolution, apparently because of suppressed recombination between mating-type chromosomes. Analysis of sequences on the mating-type chromosomes of wild collected self-fertile strains revealed high levels of genetic variability between sibling A and a nuclei. In contrast, sequences on autosomes of sibling A and a nuclei exhibited nearly complete homogeneity. Conservation of distinct haplotype combinations on A and a mating-type chromosomes in strains from diverse locations further suggested an absence of recombination over substantial periods of evolutionary time. The suppression of recombination on the N. tetrasperma mating-type chromosome, expected to ensure a high frequency of self fertility, presents an interesting parallel with, and possible model for studying aspects of, the evolution of mammalian sex chromosomes. PMID- 8725228 TI - Cold-sensitive mutations of Dictyostelium myosin heavy chain highlight functional domains of the myosin motor. AB - Dictyostelium provides a powerful environment for characterization of myosin II function. It provides well-established biochemical methods for in vitro analysis of myosin's properties as well as an array of molecular genetic tools. The absence of myosin function results in an array of phenotypes that can be used to genetically manipulate myosin function. We have previously reported methods for the isolation and identification of rapid-effect cold-sensitive myosin II mutations in Dictyostelium. Here, we report the development and utilization of a rapid method for localizing these point mutations. We have also sequenced 19 mutants. The mutations show distinct clustering with respect to three-dimensional location and biochemically characterized functional domains of the protein. We conclude that these mutants represent powerful tools for understanding the mechanisms driving this protein motor. PMID- 8725229 TI - Identification, mapping and linkage analysis of randomly amplified DNA polymorphisms in Tetrahymena thermophila. AB - Using the random amplified polymorphic DNA (RAPD) technique and exploiting the unique genetics of Tetrahymena thermophila, we have identified and characterized 40 DNA polymorphisms occurring between two inbred strains (B and C3) of this ciliated protozoan. These RAPD markers permit the PCR amplification of a DNA species using template DNA from SB1969 (B strain) but fail to do so using DNA from C3-368-5 (C3 strain). Polymorphisms were mapped to chromosomes using a panel of monosomic strains constructed by crossing B strain-derived nullisomic strains to inbred strain C3. They map to all five chromosomes and appear to be evenly distributed throughout the genome. Chromosomal groups were then analyzed for linkage using meiotic segregants; four linkage groups were identified in chromosomes 1R 2L, 3 and 5. The RAPD method appears useful for the construction of a genetic map of the Tetrahymena genome based on DNA polymorphisms. PMID- 8725230 TI - Transmission genetics of allorecognition in Hydractinia symbiolongicarpus (Cnidaria:Hydrozoa). AB - Allorecognition is ubiquitous, or nearly so, amongst colonial invertebrates. Despite the prominent role that such phenomena have played both in evolutionary theory and in on the origin of the vertebrate immune system, unambiguous data on the transmission genetics of fusibility (i.e., the ability of two individuals to fuse upon tissue contact) is lacking for any metazoan outside of the phylum Chordata. We have developed lines of the hydroid Hydractinia symbiolongicarpus (Phylum Cnidaria) inbred for fusibility and here report results of breeding experiments establishing that fusibility segregates as expected for a single locus with codominantly expressed alleles, with one shared allele producing a fusible phenotype. Surveys of fusibility in field populations and additional breeding experiments indicate the presence of an extensive allele series. PMID- 8725231 TI - Spontaneous mutational variances and covariances for fitness-related traits in Drosophila melanogaster. AB - Starting from a completely homozygous population of Drosophila melanogaster, 176 lines were derived and independently maintained by a single brother-sister mating per generation. Three fitness-related traits were considered (fecundity, egg-to pupa and pupa-to-adult viabilities). Mutational heritabilities of these traits and genetic correlations between all possible pairs were calculated from the between line divergence (codivergence), after 104-106 generations of mutation accumulation. Mutational heritabilities ranged from 0.60 x 10(-3) to 0.82 x 10( 3) and correlations from -0.11 to 0.25. These values are likely to be underestimates due to selection against deleterious mutations. The distribution of the means of the lines was asymmetric, positive for fecundity and negative for both viability components. The coefficients of asymmetry are also likely to be biased, again due to selection. Extreme lines from the two tails of the distribution were examined in detail. Homozygous line effects were all negative for viability traits but predominantly positive for fecundity, indicating the fixation of mutations with positive effects on the latter. Corresponding heterozygous line effects showed a variable degree of dominance. PMID- 8725232 TI - Age-specific patterns of genetic variance in Drosophila melanogaster. I. Mortality. AB - PETER MEDAWAR proposed that senescence arises from an age-related decline in the force of selection, which allows late-acting deleterious mutations to accumulate. Subsequent workers have suggested that mutation accumulation could produce an age related increase in additive genetic variance (VA) for fitness traits, as recently found in Drosophila melanogaster. Here we report results from a genetic analysis of mortality in 65,134 D. melanogaster. Additive genetic variance for female mortality rates increases from 0.007 in the first week of life to 0.325 by the third week, and then declines to 0.002 by the seventh week. Males show a similar pattern, though total variance is lower than in females. In contrast to a predicted divergence in mortality curves, mortality curves of different genotypes are roughly parallel. Using a three-parameter model, we find significant VA for the slope and constant term of the curve describing age-specific mortality rates, and also for the rate at which mortality decelerates late in life. These results fail to support a prediction derived from MEDAWAR's "mutation accumulation" theory for the evolution of senescence. However, our results could be consistent with alternative interpretations of evolutionary models of aging. PMID- 8725233 TI - Age-specific patterns of genetic variance in Drosophila melanogaster. II. Fecundity and its genetic covariance with age-specific mortality. AB - Under the mutation accumulation model of senescence, it was predicted that the additive genetic variance (VA) for fitness traits will increase with age. We measured age-specific mortality and fecundity from 65,134 Drosophila melanogaster and estimated genetic variance components, based on reciprocal crosses of extracted second chromosome lines. Elsewhere we report the results for mortality. Here, for fecundity, we report a bimodal pattern for VA with peaks at 3 days and at 17-31 days. Under the antagonistic pleiotropy model of senescence, it was predicted that negative correlations will exist between early and late life history traits. For fecundity itself we find positive genetic correlations among age classes > 3 days but negative nonsignificant correlations between fecundity at 3 days and at older age classes. For fecundity vs. age-specific mortality, we find positive fitness correlations (negative genetic correlations) among the traits at all ages > 3 days but a negative fitness correlation between fecundity at 3 days and mortality at the oldest ages (positive genetic correlations). For age-specific mortality itself we find overwhelmingly positive genetic correlations among all age classes. The data suggest that mutation accumulation may be a major source of standing genetic variance for senescence. PMID- 8725235 TI - A novel Drosophila Minute locus encodes ribosomal protein S13. AB - Minutes comprise > 50 phenotypically similar Drosophila mutations believed to affect ribosomal protein genes. Common traits of the Minute phenotype are short and thin bristles, slow development, and recessive lethality. To further investigate the proposed Minute to ribosomal protein correspondence, loss-of function Minute mutations were induced by P-element mutagenesis. Here, we report a previously undescribed Minute locus that maps to 32A on chromosome 2L; this Minute allele is named P{lacW}M(2)32A1 and the gene M(2)32A. Flies heterozygous for P{lacW}M(2)32A1 have a medium Minute phenotype. The gene interrupted by the P element insertion was cloned. Sequence analyses revealed that it encodes the Drosophila homologue of eukaryotic ribosomal protein S13. It is a single-copy gene and the level of RPS13 transcript is reduced to approximately 50% in P(lacW) M(2)32A1 heterozygotes. Both transcript level and phenotype are restored to wild type by remobilizing the P-element, demonstrating that the mutation is caused by insertion of the P-element construct. These results further strengthen the notion that Minutes encode ribosomal proteins and demonstrate that P-element mutagenesis is a fruitful approach to use in these studies. PMID- 8725234 TI - The nuclear protein encoded by the Drosophila neurogenic gene mastermind is widely expressed and associates with specific chromosomal regions. AB - The Drosophila neurogenic loci encode a diverse group of proteins that comprise an inhibitory signal transduction pathway. The pathway is used throughout development in numerous contexts. We have examined the distribution of the neurogenic locus mastermind protein (Mam). Mam is expressed through all germlayers during early embryogenesis, including ectodermal precursors to both neuroblasts and epidermoblasts. Mam is subsequently down-regulated within the nervous system and then reexpressed. It persists in the nervous system through late embryogenesis and postembryonically. Mam is ubiquitously expressed in wing and leg imaginal discs and is not down-regulated in sensory organ precursor cells of the wing margin or notum. In the eye disc, Mam shows most prominent expression posterior to the morphogenetic furrow. Expression of the protein during oogenesis appears limited to follicle cells. Immunohistochemical detection of Mam on polytene chromosomes revealed binding at > 100 sites. Chromosome colocalization studies with RNA polymerase and the groucho corepressor protein implicate Mam in transcriptional regulation. PMID- 8725236 TI - Mutations in the Drosophila melanogaster gene encoding S-adenosylmethionine synthetase [corrected] suppress position-effect variegation. AB - In Drosophila melanogaster, the study of trans-acting modifier mutations of position-effect variegation and Polycomb group (Pc-G) genes have been useful tools to investigate genes involved in chromatin structure. We have cloned a modifier gene, Suppressor of zeste 5 (Su(z)5), which encodes S-adenosylmethionine synthetase, and we present here molecular results and data concerning its expression in mutants and genetic interactions. The mutant alleles Su(z)5, l(2)R23 and l(2)M6 show suppression of wm4 and also of two white mutants induced by roo element insertions in the regulatory region i.e., wis (in combination with z1) and wsp1. Two of the Su(z)5 alleles, as well as a deletion of the gene, also act as enhancers of Polycomb by increasing the size of sex combs on midleg. The results suggest that Su(z)5 is connected with regulation of chromatin structure. The enzyme S-adenosylmethionine synthetase is involved in the synthesis of S adenosylmethionine, a methyl group donor and also, after decarboxylation, a propylamino group donor in the bio-synthesis of polyamines. Our results from HPLC analysis show that in ovaries from heterozygous Su(z)5 mutants the content of spermine is significantly reduced. Results presented here suggest that polyamines are an important molecule class in the regulation of chromatin structure. PMID- 8725237 TI - The alcohol dehydrogenase gene is nested in the outspread locus of Drosophila melanogaster. AB - This report describes the structure and expression of the outspread (osp) gene of Drosophila melanogaster. Previous work showed that chromosomal breakpoints associated with mutations of the osp locus map to both sides of the alcohol dehydrogenase gene (Adh), suggesting that Adh and the adjacent gene Adhr are nested in osp. We extended a chromosomal walk and mapped additional osp mutations to define the maximum molecular limit of osp as 119 kb. We identified a 6-kb transcript that hybridizes to osp region DNA and is altered or absent in osp mutants. Accumulation of this RNA peaks during embryonic and pupal periods. The osp cDNAs comprise two distinct classes based on alternative splicing patterns. The 5' end of the longest cDNA was extended by PCR amplification. When hybridized to the osp walk, the 5' extension verifies that Adh and Adhr are nested in osp and shows that osp has a transcription unit of > or = 74 kb. In situ hybridization shows that osp is expressed both maternally and zygotically. In the ovary, osp is transcribed in nurse cells and localized in the oocyte. In embryos, expression is most abundant in the developing visceral and somatic musculature. PMID- 8725238 TI - The Drosophila ash1 gene product, which is localized at specific sites on polytene chromosomes, contains a SET domain and a PHD finger. AB - The determined state of Drosophila imaginal discs depends on stable patterns of homeotic gene expression. The stability of these patterns requires the function of the ash1 gene, a member of the trithorax group. The primary translation product of the 7.5-kb ash1 transcript is predicted to be a basic protein of 2144 amino acids. The ASH1 protein contains a SET domain and a PHD finger. Both of these motifs are found in the products of some trithorax group and Polycomb group genes. We have determined the nucleotide sequence alterations in 10 ash1 mutant alleles and have examined their mutant phenotype. The best candidate for a null allele is ash1. The truncated protein product of this mutant allele is predicted to contain only 47 amino acids. The ASH1 protein is localized on polytene chromosomes of larval salivary glands at > 100 sites. The chromosomal localization of ASH1 implies that it functions at the transcriptional level to maintain the expression pattern of homeotic selector genes. PMID- 8725239 TI - Identification of immune system and response genes, and novel mutations causing melanotic tumor formation in Drosophila melanogaster. AB - We are using Drosophila as a model system for analysis of immunity and tumor formation and have conducted two types of screens using enhancer detector strains to find genes related to these processes; genes expressed in the immune system (type A; hemocytes, lymph glands and fat body) and genes increased in expression by bacterial infection (type B). For type A, tissue-specific reporter gene activity was determined. For type B, a variation of enhancer detection was devised in which beta-galactosidase is assayed spectrophotometrically with and without bacterial infection. Because of immune system involvement in melanotic tumor formation, a third type was hypothesized to be found among types A and B; genes that, when mutated, have a melanotic tumor phenotype. Enhancer detector strains (2800) were screened for type A, 900 for B, and 11 retained for further analysis. Complementation tests, cytological mapping, P-element mobilization, and determination of lethal phase and mutant phenotype have identified six novel genes, Dorothy, wizard, toto, viking, Thor and dappled, and one previously identified gene, Collagen IV. All are associated with reporter gene expression in at least one immune system tissue. Thor has increased expression upon infection. Mutations of wizard and dappled have a melanotic tumor phenotype. PMID- 8725240 TI - An integrated genetic map of the African human malaria vector mosquito, Anopheles gambiae. AB - We present a genetic map based on microsatellite polymorphisms for the African human malaria vector, Anopheles gambiae. Polymorphisms in laboratory strains were detected for 89% of the tested microsatellite markers. Genotyping was performed for individual mosquitos from 13 backcross families that included 679 progeny. Three linkage groups were identified, corresponding to the three chromosomes. We added 22 new markers to the existing X chromosome map, for a total of 46 microsatellite markers spanning a distance of 48.9 cM. The second chromosome has 57 and the third 28 microsatellite markers spanning a distance of 72.4 and 93.7 cM, respectively. The overall average distance between markers is 1.6 cM (or 1.1, 1.2, and 3.2 cM for the X, second, and third chromosomes, respectively). In addition to the 131 microsatellite markers, the current map also includes a biochemical selectable markers, Dieldrin resistance (Dl), on the second chromosome and five visible markers, pink-eye (p) and white (w) on the X, collarless (c) and lunate (lu) on the second, and red-eye (r) on the third. The cytogenetic locations on the nurse cell polytene chromosomes have been determined for 47 markers, making this map an integrated tool for cytogenetic, genetic, and molecular analysis. PMID- 8725241 TI - Integrated genetic map of Anopheles gambiae: use of RAPD polymorphisms for genetic, cytogenetic and STS landmarks. AB - Randomly amplified polymorphic DNA (RAPD) markers have been integrated in the genetic and cytogenetic maps of the malaria vector mosquito, Anopheles gambiae. Fifteen of these markers were mapped by recombination, relative to microsatellite markers that had been mapped previously. Thirty-four gel-purified RAPD bands were cloned and sequenced, generating sequence tagged sites (STSs) that can be used as entry points to the A. gambiae genome. Thirty one of these STSs were localized on nurse cell polytene chromosomes through their unique hybridization signal in in situ hybridization experiments. Five STSs map close to the breakpoints of polymorphic inversions, which are notable features of the Anopheles genome. The usefulness and limitations of this integrated mosquito map are discussed. PMID- 8725242 TI - Single and double infections with Wolbachia in the parasitic wasp Nasonia vitripennis: effects on compatibility. AB - Wolbachia are cytoplasmically inherited bacteria responsible for reproductive incompatibility in a wide range of insects. There has been little exploration, however, of within species Wolbachia polymorphisms and their effects on compatibility. Here we show that some strains of the parasitic wasp Nasonia vitripennis are infected with two distinct bacterial strains (A and B) whereas others are singly infected (A or B). Double and single infections are confirmed by both PCR amplification and Southern analysis of genomic DNA. Furthermore, it is shown that prolonged larval diapause (the overwintering stage of the wasp) of a double-infected strain can lead to stochastic loss of one or both bacterial strains. After diapause of a double-infected line, sublines were produced with AB, A only, B only or no Wolbachia. A and B sublines are bidirectionally incompatible, whereas males from AB lines are unidirectionally incompatible with females of A and B sublines. Results therefore show rapid development of bidirectional incompatibility within a species due to segregation of associated symbiotic bacteria. PMID- 8725243 TI - Isolation of Arabidopsis mutants with enhanced disease susceptibility by direct screening. AB - To discover which components of plant defense responses make significant contributions to limiting pathogen attack, we screened a mutagenized population of Arabidopsis thaliana for individuals that exhibit increased susceptibility to the moderately virulent bacterial pathogen Pseudomonas syringae pv. maculicola ES4326 (Psm ES4326). The 12 enhanced disease susceptibility (eds) mutants isolated included alleles of two genes involved in phytoalexin biosynthesis (pad2, which had been identified previously, and pad4, which had not been identified previously), two alleles of the previously identified npr1 gene, which affects expression of other defense genes, and alleles of seven previously unidentified genes of unknown function. The npr1 mutations caused greatly reduced expression of the PR1 gene in response to PsmES4326 infection, but had little effect on expression of two other defense genes, BGL2 and PR5, suggesting that PR1 expression may be important for limiting growth of PsmES4326. While direct screens for mutants with quantitative pathogen-susceptibility phenotypes have not been reported previously, our finding that mutants isolated in this way include those affected in known defense responses supports the notion that this type of screening strategy allows genetic dissection of the roles of various plant defense responses in disease resistance. PMID- 8725244 TI - Genetic map of diploid wheat, Triticum monococcum L., and its comparison with maps of Hordeum vulgare L. AB - A genetic map of diploid wheat, Triticum monococcum L., involving 335 markers, including RFLP DNA markers, isozymes, seed storage proteins, rRNA, and morphological loci, is reported. T. monococcum and barley linkage groups are remarkably conserved. They differ by a reciprocal translocation involving the long arms of chromosomes 4 and 5, and paracentric inversions in the long arm of chromosomes 1 and 4; the latter is in a segment of chromosome arm 4L translocated to 5L in T. monococcum. The order of the markers in the inverted segments in the T. monococcum genome is the same as in the B and D genomes of T. aestivum L. The T. monococcum map differs from the barley maps in the distribution of recombination within chromosomes. The major 5S rRNA loci were mapped on the short arms of T. monococcum chromosomes 1 and 5 and the long arms of barley chromosomes 2 and 3. Since these chromosome arms are colinear, the major 5S rRNA loci must be subjected to positional changes in the evolving Triticeae genome that do not perturb chromosome colinearity. The positional changes of the major 5S rRNA loci in Triticeae genomes are analogous to those of the 18S-5.8S-26S rRNA loci. PMID- 8725245 TI - Identification and high-density mapping of gene-rich regions in chromosome group 5 of wheat. AB - The distribution of genes and recombination in the wheat genome was studied by comparing physical maps with the genetic linkage maps. The physical maps were generated by mapping 80 DNA and two phenotypic markers on an array of 65 deletion lines for homoeologous group 5 chromosomes. The genetic maps were constructed for chromosome 5B in wheat and 5D in Triticum tauschii. No marker mapped in the proximal 20% chromosome region surrounding the centromere. More than 60% of the long arm markers were present in three major clusters that physically encompassed < 18% of the arm. Because 48% of the markers were cDNA clones and the distributions of the cDNA and genomic clones were similar, the marker distribution may represent the distribution of genes. The gene clusters were identified and allocated to very small chromosome regions because of a higher number of deletions in their surrounding regions. The recombination was suppressed in the centromeric regions and mainly occurred in the gene-rich regions. The bp/cM estimates varied from 118 kb for gene-rich regions to 22 Mb for gene-poor regions. The wheat genes present in these clusters are, therefore, amenable to molecular manipulations parallel to the plants with smaller genomes like rice. PMID- 8725246 TI - Confidence intervals in QTL mapping by bootstrapping. AB - The determination of empirical confidence intervals for the location of quantitative trait loci (QTLs) was investigated using simulation. Empirical confidence intervals were calculated using a bootstrap resampling method for a backcross population derived from inbred lines. Sample sizes were either 200 or 500 individuals, and the QTL explained 1, 5, or 10% of the phenotypic variance. The method worked well in that the proportion of empirical confidence intervals that contained the simulated QTL was close to expectation. In general, the confidence intervals were slightly conservatively biased. Correlations between the test statistic and the width of the confidence interval were strongly negative, so that the stronger the evidence for a QTL segregating, the smaller the empirical confidence interval for its location. The size of the average confidence interval depended heavily on the population size and the effect of the QTL. Marker spacing had only a small effect on the average empirical confidence interval. The LOD drop-off method to calculate empirical support intervals gave confidence intervals that generally were too small, in particular if confidence intervals were calculated only for samples above a certain significance threshold. The bootstrap method is easy to implement and is useful in the analysis of experimental data. PMID- 8725247 TI - Constraints on allele size at microsatellite loci: implications for genetic differentiation. AB - Microsatellites are promising genetic markers for studying the demographic structure and phylogenetic history of populations. We present theoretical arguments indicating that the usefulness of microsatellite data for these purposes may be limited to a short time perspective and to relatively small populations. The evolution of selectively neutral markers is governed by the interaction of mutation and random genetic drift. Mutation pressure has the inherent tendency to shift different populations to the same distribution of alleles. Hence, mutation pressure is a homogenizing force, and population divergence is caused by random genetic drift. In case of allozymes or sequence data, the diversifying effect of drift is typically orders of magnitude larger than the homogenizing effect of mutation pressure. By a simple model, we demonstrate that the situation may be different for microsatellites where mutation rates are high and the range of alleles is limited. With the help of computer simulations, we investigate to what extent genetic distance measures applied to microsatellite data can nevertheless yield useful estimators for phylogenetic relationships or demographic parameters. We show that predictions based on microsatellite data are quite reliable in small populations, but that already in moderately sized populations the danger of misinterpretation is substantial. PMID- 8725248 TI - A population genetic study of the evolution of SINEs. II. Sequence evolution under the master copy model. AB - A transient population genetic model of SINE (short interspersed repetitive element) evolution assuming the master copy model is theoretically investigated. Means and variances of consensus frequency of nucleotides, nucleotide homozygosity, and the number of shared differences that are considered to have caused by mutations occurring in the master copy lineages are computed. All quantities investigated are shown to be monotone functions of the duration of the expansion period. Thus, they can be used to estimate the expansion period although their sampling variances are generally large. Using the theoretical results, the Sb subfamily of human Alu sequences is analyzed. First, the expansion period is estimated from the observed mean and variance of homozygosity. The expansion period is shown to be short compared to the time since the end of the expansion of the subfamily. However, the observed number of the shared differences is more than twice that expected under the master copy model with the estimated expansion period. Alternative models including that with multiple master copy loci to explain this observation are discussed. PMID- 8725249 TI - The probability distribution of the amount of an individual's genome surviving to the following generation. AB - The probability that at least p% of an individual's genome is passed on collectively to his children is calculated. With data availability the consideration of the chromosome as a whole rather than discrete loci becomes of increasing practical importance. Assuming the genomic continuum model, which allows for recombination, the crossover process in a chromosome pedigree is viewed as a continuous-time Markov random walk on the vertices of a hypercube with time parameter map distance along the chromosome. The desired probability corresponds to the probability of sojourn times of the process in a small set of vertices, which are well approximated via the Poisson clumping heuristic. Results are given for the human genome. It is very likely that an individual with at least four children passes on at least 90% of his genome. There exists no "equivalent" number of independently segregating loci for this distribution. PMID- 8725250 TI - Changes in structure, mechanics, and insulin-like growth factor-related gene expression in the lungs of newborn rats exposed to air or 60% oxygen. AB - Exposure of neonatal rats to > or = 95% O2 for 2 wk, a widely used model of oxidant/antioxidant interactions in neonatal lung injury, results in arrested lung growth without the dysplastic lesions observed in chronic human neonatal lung injury. To determine whether dysplastic lung cell growth would be seen at lesser O2 concentrations, we exposed newborn rats to either 95% O2 for 1 wk followed by 60% O2 for 1 wk, or to 60% O2 for 2 wk. Exposure to 95% O2 for 1 wk profoundly inhibited lung DNA synthesis. Recovery of synthesis did not occur during the 2nd wk in 60% O2, nor were areas of dysplastic growth evident in lung tissue. In contrast, a continuous 2-wk exposure to 60% O2 resulted in a slight increase in lung weight with a significant reduction in lung volume over a range of inflation pressures. Also seen was an overall, but inhomogeneous, reduction in lung cell DNA synthesis. A preliminary analysis of affected cell types suggested that inhibition of DNA synthesis affected endothelial cells more than interstitial cells, whereas DNA synthesis increased in type II pneumocytes. Areas of reduced DNA synthesis were interspersed with patchy areas of parenchymal thickening and active DNA synthesis. These areas of parenchymal thickening, but not other areas, had increased immunoreactive IGF-I and the type I IGF receptor. These data are consistent with a direct effect of O2 on growth factor and growth factor receptor expression in causing dysplastic lung cell growth in chronic neonatal lung injury. PMID- 8725251 TI - Localization and developmental expression of surfactant proteins D and A in the respiratory tract of the mouse. AB - Surfactant protein D (SP-D) is synthesized and secreted by pulmonary epithelial cells. Like surfactant protein A (SP-A), SP-D is a collagen-like glycoprotein belonging to the "collectin" class of C-type lectins that may play an important role in pulmonary host defense. To begin studies on SP-D gene regulation and function using the mouse as an animal model, we identified the cellular sites of SP-D gene expression in adult mouse lung and trachea and characterized the developmental expression of SP-D mRNA in murine fetal and newborn lungs. We compared these findings with similar studies for murine SP-A, which has an established role in surfactant function and metabolism and a probable role in pulmonary host defense. SP-D mRNA and protein were readily detected by in situ hybridization and immunocytochemistry in alveolar type II and nonciliated bronchiolar epithelial cells of the lung, as well as in cells of the tracheal epithelium and tracheal submucosal glands of the adult mouse. Although SP-A mRNA and protein were also localized to alveolar and nonciliated bronchiolar epithelial cells of the murine lung, there was no detectable labeling for either SP-A mRNA or protein in the murine trachea. Expression of murine SP-D mRNA was first detected by Northern blot analysis on d 16 of gestation in timed-pregnant mice, with an average gestational period of 17 d, and this increased dramatically before birth and during the immediate postnatal period. The developmental expression of murine SP-A mRNA paralleled that of SP-D except that there was a small decrease in mRNA content on postnatal d 5. These studies provide the first description of the cellular distribution and developmental expression of SP-D in mouse lung, which will be important for interpreting future studies of SP-D gene expression in transgenic animal models. In addition, these studies provide the first documentation that, unlike SP-A, SP-D is synthesized not only in the lung but also in submucosal glands of the trachea. PMID- 8725252 TI - A synthetic segment of surfactant protein A: structure, in vitro surface activity, and in vivo efficacy. AB - Surfactant protein A (SP-A) is a 248-residue, water-soluble, lipid-associating protein found in lung surfactant. Analysis of the amino acid sequence using the Eisenberg hydrophobic moment algorithm predicts that the SP-A segment spanning residues 114-144 has high hydrophobic moments, typical of lipid-associating amphipathic domains. The secondary structure, in vitro surface activity and in vivo lung activity of this SP-A sequence were studied with a 31-residue synthetic peptide analog (A114-144). Analysis of the secondary structure using circular dichroism and Fourier transform infrared spectroscopy indicated association with lipid dispersions and a dominant helical content. Surface activity measurements of A114-144 with surfactant lipid dispersions and the hydrophobic surfactant proteins B and C (SP-B/C) showed that A114-144 enhances surface activity under conditions of dynamic compression and respreading on a Langmuir/Wilhelmy surface balance. Synthetic surfactant dispersions containing A114-144 improved lung compliance in spontaneously breathing, 28-d premature rabbits to a greater degree than surfactant dispersions with synthetic SP-B/C and synthetic surfactant lipids alone. These observations indicate that inclusion of A114-144 may improve synthetic preparations currently used for surfactant replacement therapy. PMID- 8725253 TI - Comparative effects of some serum components and proteolytic products of fibrinogen on surface tension-lowering abilities of beractant and a synthetic peptide containing surfactant KL4. AB - The serum components of C-reactive protein, lysophosphati-dylcholine, fibrinogen, and fibrinogen proteolytic products have been shown to reduce surface tension lowering abilities of lung surfactant. The inhibitory effects of these serum components were compared among four different surfactants: natural lung surfactant, a phospholipid mixture that had no surfactant proteins, KL4 surfactant which has a synthetic surfactant protein B (SP-B)-like peptide, and beractant (BER) which has both SP-B and SP-C. The pulsating bubble surfactometer was used to measure the surface tension of these surfactants after the addition of inhibitors. Inhibition of BER and KL4 surfactant was observed with some serum components within 1 min of pulsation, but was reversed after 3 min of pulsation for KL4 surfactant and to a lesser extent with BER. The surface tension of phospholipid mixture alone was significantly increased and did not improve with further pulsations. Natural lung surfactant was least inhibited and was affected only at very high fibrinogen concentrations (5 mg/mL). At identical concentrations of these inhibitors, KL4 surfactant was inhibited less compared with BER. We conclude that the response of a lung surfactant to inhibitory agents may depend on the presence or absence of surfactant-related protein(s) in the surfactant and the concentration of exogenous surfactant used. KL4 surfactant, which has a synthetic peptide in lieu of SP-B, resists inhibition to these serum components more than BER at similar phospholipid concentrations. PMID- 8725254 TI - Surfactant aerosol treatment of respiratory distress syndrome in the spontaneously breathing premature rabbit. AB - Surfactant deficiency in premature neonates is a major factor in the development of respiratory distress syndrome (RDS), which is still a significant cause of mortality and morbidity. The aim of this study was to test a noninvasive method of administering surfactant as treatment for RDS. The animal model used was the premature neonatal rabbit of 27-d gestation (full-term being 31 d) primed with an initial oropharyngeal dose of surfactant. The animals were divided into three groups that received either no supplemental surfactant (n = 20), undried nebulized surfactant (n = 21), or dried nebulized surfactant (n = 24). Drying of the surfactant solution was undertaken to create a hygroscopic aerosol that would facilitate surfactant deposition in the lower respiratory tract. The group treated with dried surfactant aerosol showed superior survival (66.7%) and less evidence of RDS. The control and undried aerosol groups each had similar low survival rates (23.8 and 45.0%, respectively). The results indicate that a dried, hygroscopic aerosol is an effective means of administration of surfactant to spontaneously breathing premature rabbit neonates. PMID- 8725255 TI - Leakage of protein into lungs of preterm ventilated rabbits is correlated with activation of clotting, complement, and polymorphonuclear leukocytes in plasma. AB - We investigated whether leakage of protein in lungs of preterm ventilated rabbits of 28- and 29-d gestational age is correlated with activation of clotting, complement, and polymorphonuclear leukocytes (PMN) in plasma. We found signs of systemic activation of clotting, complement and PMN in ventilated 28-d gestational age rabbits, as indicated, respectively, by increased median plasma fibrin monomer concentrations (83 versus 40% of normal adult rabbit plasma in nonventilated 28-d gestational age rabbits, p < 0.01), decreased median plasma CH50 activity (112 versus 122 U/L in nonventilated 28-d gestational age rabbits, p < 0.05), and increased median plasma beta-glucuronidase concentrations (159 versus 97% of maximal activated adult rabbit plasma in nonventilated 28-d gestational age rabbits p < 0.05). We did not find signs of systemic activation in the ventilated 29-d gestational age group. Higher median total protein concentrations in alveolar wash of the ventilated 28-d gestational age rabbits (2.7 versus 1.3 mg/mL in the nonventilated rabbits. p < 0.01) indicated protein leakage into the lungs, and this protein leakage was more pronounced in the lungs of ventilated 28-d gestational age rabbits than in those of ventilated 29-d gestational age rabbits (2.1 mg/mL, p < 0.01). The total protein concentration in the alveolar wash of all 28-d gestational age rabbits was correlated with the concentration of fibrin monomers (p = 0.51, p = 0.035) and beta-glucuronidase (p = 0.61, p = 0.011), and the CH50 activity (p = -0.73, p = 0.002) in plasma. We conclude that leakage of protein in lungs of preterm ventilated rabbits of 28-d gestational age is correlated with activation of clotting, complement, and PMN in plasma. This activation process may contribute to lung injury by intravascular and intraalveolar deposition of fibrin and formation of proteinaceous edema. PMID- 8725256 TI - Undetectable interleukin (IL)-10 and persistent IL-8 expression early in hyaline membrane disease: a possible developmental basis for the predisposition to chronic lung inflammation in preterm newborns. AB - We are interested in determining whether premature birth alters expression of counterregulatory cytokines which modulate lung inflammation. Production of proinflammatory cytokines tumor necrosis factor alpha. IL-1 beta, and IL-8 is regulated in part by the antiinflammatory cytokine IL-10. For preterm newborns with hyaline membrane disease, deficiencies in the ability of lung macrophages to express antiinflammatory cytokines may predispose to chronic lung inflammation. We compared the expression of pro- and antiinflammatory cytokines at the mRNA and protein level in the lungs of preterm and term newborns with acute respiratory failure from hyaline membrane disease or meconium aspiration syndrome. Four sequential bronchoalveolar lavage (BAL) samples were obtained during the first 96 h of life from all patients. All patients rapidly developed an influx of neutrophils and macrophages. Over time, cell populations in both groups became relatively enriched with macrophages. The expression of proinflammatory cytokine mRNA and/or protein was present in all samples from both patient groups. In contrast, IL-10 mRNA was undetectable in most of the cell samples from preterm infants and present in the majority of cell samples from term infants. IL-10 concentrations were undetectable in lavage fluid from preterm infants with higher levels in a few of the BAL samples from term infants. These studies demonstrate that 1) IL-10 mRNA and protein expression by lung inflammatory cells is related to gestational age and 2) during the first 96 h of life neutrophil cell counts and IL-8 expression decrease in BAL from term infants, but remain unchanged in BAL samples from preterm infants. PMID- 8725257 TI - Interleukin-6 expression in cord blood of patients with clinical chorioamnionitis. AB - The objective of this study was to define whether IL-6 is an early marker of infection in the newborn. To correlate the occurrence of clinical chorioamnionitis with the levels of IL-6 expression in neonates, IL-6 was measured in cord plasma by ELISA and in mononuclear cells by reverse transcriptase-PCR before and after mitogenic stimulation. Eight neonates were included in each of the following four groups: elective cesarean section, uncomplicated normal spontaneous vaginal delivery, delivery after prolonged rupture of amniotic membranes with no evidence of chorioamnionitis, and delivery with evidence of chorioamnionitis. All 32 neonates were clinically well after delivery, and all 16 babies with prolonged rupture of membranes or clinical chorioamnionitis had negative blood cultures. Elevated IL-6 levels were found only in neonates born to mothers with chorioamnionitis (119.7 +/- 33.5 pg/mL versus 2.71 +/- 0.59 pg/mL, p < 0.005). Mononuclear cells from five of these neonates expressed no IL-6 mRNA in vivo despite elevated levels of IL-6 in their cord plasma. Cord blood mononuclear cells from healthy term babies were capable of synthesizing IL-6 in vitro in response to stimulation with bacterial lipopolysaccharide. These results suggest that IL-6 levels in cord plasma increased with clinical chorioamnionitis, despite the lack of evidence of infection in the neonates. Therefore, we conclude that, although a high level of IL-6 may be a good marker of chorioamnionitis, it may not be a specific marker of infection in the newborn. PMID- 8725258 TI - Distribution of the predominant hepatitis B virus precore variants in hepatitis B e antigen-positive children and their effect on treatment response. AB - Little is known about the distribution of precore hepatitis B virus mutants and their influence on the outcome of interferon therapy in children with chronic hepatitis B. In this study, serum samples were analyzed from 60 children with chronic hepatitis B e antigen+ (HBeAg+) hepatitis. Fifty-two of these children underwent different interferon trials, and a second serum sample was taken from 25 of them at the end of therapy. Fifty-six of the original 60 children were simultaneously infected by wild-type and precore mutant hepatitis B virus variants. The remaining four children were infected by the wild type alone. In 50/56 of children with a mixed viral population, the wild-type variant comprised more than 50% of the total viremia. With respect to the influence of precore variants on the outcome of interferon treatment, the prevalence of mixed viral population was similar in responders and nonresponders (96 versus 88%, respectively). However, precore mutant variants were prevalent (> 50% of the viral population) in 21% of the nonresponders, but in none of the responder children (p < 0.05). Viremia levels were significantly higher in nonresponder than in responder children (p < 0.05). No substantial changes in the prevalence of mutants were observed throughout the interferon therapy. In conclusion, mixed viral infection is found in a high percentage of children with chronic B HBeAg+ hepatitis. Response to interferon therapy does not seem to be related to the presence of hepatitis B virus precore mutants, but rather to the levels of viremia. PMID- 8725259 TI - Intestinal permeability in exocrine pancreatic insufficiency due to cystic fibrosis or chronic pancreatitis. AB - Disturbances of the intestinal integrity, reflected by an increased intestinal permeability, are reported in cystic fibrosis (CF). Controversy exists whether the increased intestinal permeability is due to CF itself or a consequence of the concomitant exocrine pancreatic insufficiency (PI). We measured intestinal permeability by the sugar absorption test in 32 PI patients: 20 CF-PI, 12 nonCF PI with chronic pancreatitis, and 50 controls. In the sugar absorption test, the lactulose/mannitol ratio is measured in 5-h urine samples after oral ingestion of a solution of lactulose and mannitol, hyperosmolar by the addition of sucrose. The lactulose/mannitol ratio was increased in both CF-PI and nonCF-PI versus controls (p < 0.0001). In CF, the L/M ratio and permeability for lactulose and mannitol did not change by increasing pancreatic enzyme supplementation by 30-50% for 2 wk (p = 0.74, p = 0.97, p = 0.74, respectively) nor by decreasing the osmolarity of the test solution by 75% (p = 0.24, p = 0.10, p = 0.39, respectively). We conclude that an increased intestinal permeability in CF is probably a consequence of PI and is not related to the dose of pancreatic enzyme supplementation nor the osmolarity of the test solution. The increase is due to an increased permeability for lactulose which might point toward a defect in the tight junctions of the villi and/or crypts. The cause of the increased intestinal permeability in the presence of PI is still unclear. An increased intestinal permeability points toward an impaired functional integrity of the small bowel, which may contribute to gastrointestinal dysfunction in CF. PMID- 8725260 TI - Insulin-like growth factor I, its binding proteins 1 and 3, and growth hormone binding protein in children and adolescents with insulin-dependent diabetes mellitus: clinical implications. AB - Values of IGF-I after extraction, its binding proteins, and the high affinity GH binding protein (BP) are not well established in pediatric patients with insulin dependent diabetes mellitus (IDDM). We report data for IGF-I, IGFBP-1, and -3, and GHBP in 92 Spanish children with IDDM, separated according to pubertal stage: prepubertal (n = 49); pubertal onset (n = 17); mid-puberty (n = 17), and complete puberty (n = 9), as well as to metabolic control (HbA1 < 9% or > or = 9%). IGF-I levels in IDDM patients increased throughout development (p < 0.001), but were diminished at every developmental stage when compared with marched control subjects. IGF-I concentrations showed a negative correlation with the degree of metabolic control, in particular during the prepubertal stage of development. A negative correlation (r = -0.22; p < 0.005) between IGF-I concentrations and HbA1 was found. Serum IGFBP-I levels diminish during maturation in diabetic patients (p < 0.001). However, IDDM patients have significantly higher levels of IGFBP-1 than control subjects at every stage of development, and IDDM patients with inadequate metabolic control exhibit even greater differences when compared with matched control subjects. A positive correlation (r = 0.22; p < 0.005) between IGFBP-1 concentrations and HbA1 was found. IGFBP-3 serum levels were similar to those observed in normal subjects, and no correlation was observed in relation to the metabolic control. In IDDM patients, GHBP levels change significantly during maturation, as they do in normal control subjects; however, significantly lower GHBP levels were found in prepubertal and pubertal IDDM patients. GHBP levels were independent of metabolic control, although a tendency toward lower levels of GHBP was seen when HbA1 levels increased. We suggest that a partial GH resistance syndrome exists in IDDM patients, and this may be related to the metabolic control. Hence, the biochemical markers measured here may be of value in evaluating the smaller pubertal growth spurt in diabetic patients. PMID- 8725261 TI - Effects of early parenting on growth and development in a small primate. AB - We report here a study of the impact of caregiver-infant relationships on physical growth and behavioral development in a small primate, the common marmoset. Somatic growth was assessed from measurements of body weight, knee-heel length, head-tail length, head circumference, and pudendal pad width in females or testis volume in males obtained from unanesthetized monkeys. Behavioral information was gathered by focal animal samples for discrete rearing behaviors. Our data suggest that the frequency of positive parental behaviors during infancy is correlated with stature when the monkeys reach 10 and 20 wk of age. Furthermore, we found that juveniles that were mistreated by their parents during infancy were smaller in body weight, knee-heel length, and head-tail length, and they demonstrated abnormal social behavior. Finally, to address whether the apparent decreased growth observed in the young animals that had experienced negative parenting was also associated with alterations in hypothalamic-pituitary adrenal (HPA) axis function, we examined the plasma ACTH and cortisol responses to synthetic ovine corticotropin-releasing hormone (oCRH) in these animals. We found that the incremental cortisol response to exogenous oCRH was significantly lower in the young adults that had experienced negative parenting during infancy compared with those who had nonabusive parents, indicating altered hypothalamic pituitary-adrenal axis function in these animals. Our findings suggest that the quality of parental care influences later growth and behavior in the young marmoset. PMID- 8725262 TI - Pulsatile release and circadian rhythms of thyrotropin and prolactin in children with growth hormone deficiency. AB - We have measured mean concentrations and have appraised the pulsatile nature of thyrotropin (TSH) and prolactin (PRL) release in children with classical GH deficiency (GHD; n = 4) and neurosecretory GH dysfunction (NSD; n = 4) and have compared the results with those obtained in children with constitutional delay (control; n = 4). Blood samples were obtained at 20-min intervals for 24 h. Pulse analysis of TSH and PRL was undertaken using the Cluster pulse detection algorithm. Circadian rhythmicity of TSH and PRL was assessed using cosinor analysis. The mean 24-h concentration of GH in the control subjects was significantly higher than that obtained in the GHD and NSD groups. With regard to TSH, the mean serum concentration in the GHD and NSD group were higher than that of the control subjects. This augmentation reflects TSH pulses of large amplitude and area, and a higher interpulse valley mean rather than a difference in peak number or peak duration. No differences in mean PRL concentration or characteristics of PRL pulses were found between the control and GHD and NSD subjects. When the 24 h data sets were divided into day (0800-2000 h) and night (2000-0800 h), the mean nighttime TSH concentration was higher than the daytime concentration in the control, GHD, and NSD groups. Although there were no day versus night differences in TSH pulse frequency in either group, peak amplitude, area, and interpulse valley means were increased during the night in the control group, and peak area, duration, and amplitude mean in the NSD group. The nighttime mean PRL concentrations in the control, GHD, and NSD subjects were higher than those found during the day. This increase was accounted for by increases in PRL peak amplitude, area in the control group, and peak area, amplitude, and interpulse valley mean in the GHD and NSD groups. Cosinor analysis of the 24-h TSH and PRL data revealed clear circadian rhythmicity in all groups of subjects. These data suggest that GHD and NSD are associated with an increase in pulsatile TSH secretion due to an increase in pulse amplitude and interpulse valley mean. PMID- 8725264 TI - Prevention of trauma-induced neurodegeneration in infant rat brain. AB - Recent evidence implicates the endogenous excitatory neurotransmitters, glutamate (Glu) and aspartate, in the pathophysiology of traumatic injury in the adult CNS, but it is not known whether similar excitotoxic mechanisms mediate traumatic injury in the immature CNS. Therefore, we developed a model of brain contusion injury in infant rats and used this model to study the nature and evolution of the acute cytopathologic changes and to evaluate the ability of Glu receptor antagonists to protect the immature brain against such changes. Seven-day-old rat pups were subjected to contusion injury and were killed 0, 0.5, 1, 2, 4, and 6 h later for histologic evaluation of the brain. Physical tearing of the dura and minor disruption of underlying brain tissue was noted at 0 h. At 30 min a discrete zone of neuronal necrosis began to appear at the border of the trauma site; this zone progressively expanded over a period of 4 h. The cytopathologic changes closely resembled the type of changes Glu is known to cause; these changes consisted of swollen dendrites, degenerating neurons with pyknotic nuclei and markedly swollen cytoplasm, and dark cells with vacuolated cytoplasm. The noncompetitive N-methyl-D-aspartate (NMDA) antagonist, dizocilpine maleate, when administered 30 min before or 1 h after trauma, significantly attenuated the lesion. The competitive NMDA antagonist, 3-((-2)-carboxypiperazine-4-yl)-propyl-1 phosphonate, was also neuroprotective. The alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionate/kainate receptor antagonist 2,3-dihydro-6-nitro-7-sulfamoyl benzo(f)quinoxaline did not significantly suppress the lesion when given as three treatments (30 mg/kg each) 30 min before plus 15 and 75 min after the insult. These findings suggest that traumatic injury in the infant rat brain is mediated by endogenous excitotoxins (Glu and aspartate) acting at NMDA receptors and can be substantially mitigated by timely treatment with NMDA receptor antagonists. PMID- 8725263 TI - Modified hormonal effects on fat metabolism after severe head injury in children. AB - Previous studies within our research group have indicated that the hormonal influences on whole body energy expenditure may be modified in severely head injured children. The aim of this study was to examine plasma concentrations of nonesterified fatty acids (NEFA) and the hormonal and metabolic mediators which influence these to determine whether there is a similarly modified effect on fat metabolism. A total of 64 serial measurements were made in 21 fasting severely head-injured children aged 2-15 y (Glasgow Coma Score < or = 8) who were receiving neurointensive care. Circulating NEFA, ketone bodies, and lactate concentrations were analyzed using microenzymatic or electrochemical techniques. Plasma concentrations of adrenaline and insulin were measured using radioenzymatic and RIA techniques, respectively. Net fat oxidation rates were determined using indirect calorimetry. Plasma NEFA concentrations showed a significant positive relationship with both net fat oxidation rates (p = 0.02) and log ketone body concentrations (p = 0.008), indicating that NEFA concentrations were significantly related with utilization. When compared with reference values for normal resting adults, 59 (92%) adrenaline measurements were elevated, whereas only 8 (12%) NEFA values lay above the reference range. Surprisingly, between children, there was a significant negative relationship between NEFA and adrenaline concentrations, even after allowing for the effects of insulin and lactate (p = 0.015). Both plasma NEFA and adrenaline concentrations were significantly related with Glasgo Coma Score (p = 0.04, p = 0.007, respectively), the most severely injured children having the lowest NEFA and highest adrenaline concentrations. The mechanisms underlying these metabolic changes may be related to the severity of head injury and may involve changes in triglyceride/NEFA cycling and/or peripheral effects on adrenergic receptors. If children are to be treated effectively after trauma, it is important to discover the mechanism of these changes which must reflect a fundamental alteration in metabolism. PMID- 8725265 TI - Differential effects of 3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors on the development of myopathy in young rats. AB - HMG-CoA reductase inhibitors (statins), cholesterol-lowering drugs that have not been approved for use in children and adolescents, may cause myopathy as a side effect. We compared the effects of three statins (simva-, prava- and lovastatin) in young rats to determine whether skeletal muscle of young animals is more susceptible than that of adults. We also evaluated whether the type of statin (lipophilic versus hydrophilic) determines the degree of muscle damage. Administration via chow of simvastatin (15 mg/kg of body weight/d) and lovastatin (43-55 mg/kg of body weight/d), both lipophilic, caused stunted growth, high creatine kinase (CK) activity in plasma, and severe myopathy. Statin doses that caused damage were much lower for young rats than for adults. Pravastatin (8-55 mg/kg of body weight/d), a hydrophilic drug, caused none of these symptoms. Histologic analysis of hind paw muscles of simvastatin-and lovastatin-treated rats showed abundant signs of damage (hypercontraction, fiber necrosis) in the extensor digitorum longus, correlating with the symptoms noted above. No cellular infiltrates were seen at the onset, pointing to a noninflammatory myopathy. Pravastatin-treated rats never showed signs of myopathy. Impaired DNA synthesis may explain why muscle toxicity is seen at lower doses in young, rapidly developing rats than in adult animals. The differences in muscle damage between the statins may be attributed to differences in lipophilicity and thus in tissue selectivity. Our results can be important when considering drug therapy in young patients with inherited lipoprotein disorders. PMID- 8725266 TI - Tyrosine uptake and regional brain monoamine metabolites in a rat model resembling congenital hyperammonemia. AB - Hyperammonemia found in congenital disorders has a toxic effect on the central nervous system. Disturbances of brain neurotransmitter metabolism have been proposed, such as an increased transport of tryptophan into the brain and an increased flux through the serotonin pathway. Results concerning the catecholamine pathway are, however, contradictory. We therefore studied whether hyperammonermia increases brain uptake of the neurotransmitter precursor amino acid tyrosine and whether these changes affect the concentration of neurotransmitters and their metabolites in different brain areas (frontal cortex, caudatus-putamen, thalamus, hypothalamus, hippocampus/substantia nigra, brainstem) of rats made hyperammonemic with urease. The brain uptake of tyrosine was measured in the forebrain, brainstem, and cerebellum. The brain areas were analyzed for dopamine, 3,4-hydroxyphenylacetic acid; homovanillic acid, norepinephrine, and vanillylmandelic acid. The brain uptake index of tyrosine was increased in the forebrain and brainstem of the hyperammonemic rats with concomitantly elevated concentrations in the forebrain of tyrosine, phenylalanine, and tryptophan. The homovanillic acid content was significantly increased in the hypothalamus, hippocampus/substantia nigra and brainstem. The concentrations of norepinephrine, dopamine, and 3, 4-hydroxyphenylacetic acid were not significantly changed. Vanillylmandellic acid was decreased in the caudatus-putamen, thalamus, and hypothalamus. The data indicate an undisturbed neurotransmitter synthesis and, taken with the augmented tyrosine uptake at the blood-brain barrier, an increased flux through the dopamine pathway. These changes observed in the hyperammonemic animal model could contribute to the understanding of the pathogenic mechanisms and offer an explanation for the neuropsychiatric disturbances observed in children with congenital hyperammonemia. PMID- 8725267 TI - Duration of action and tissue distribution of zinc protoporphyrin in neonatal rats. AB - Zinc protoporphyrin IX (ZnPP) has been shown to inhibit heme oxygenase (HO) activity effectively in vivo and has potential in the treatment of neonatal jaundice. Because this is a transitional or temporary condition lasting only several days, an effective chemopreventive agent with a relatively short duration of action would be desirable for the treatment of severe neonatal jaundice. To determine the effective duration of action of ZnPP, we administered either 40 nmol/g of body weight ZnPP or 5 microL/g body weight diluent intraperitoneally to neonatal rats 24-36 h after birth. Between 0 and 21 d after ZnPP dosing, the duration of action was investigated through measurements of serum bilirubin and hepatic and splenic HO inhibition, which were correlated to measurements of ZnPP distribution. Significant (p < 0.05) hepatic HO inhibition, ranging from 27 to 51%, was observed in the liver between 1 and 4 d after dosing, concurrent with a 23-28% reduction in serum bilirubin levels, and was associated with ZnPP tissue concentrations of 27-38 nmol/g. Splenic HO was not inhibited measurably by the much lower concentrations of ZnPP found in the spleen (2.8-20.1 nmol/g) between 0 and 21 d after dosing. Furthermore, HO isoform 1 (HO-1) induction was apparently not a confounding factor in the duration of action of ZnPP, because the modest increases in HO-1 protein levels were not sustained longer than 24 h after ZnPP administration. Our findings demonstrated that the duration of action of ZnPP in neonatal rats is less than 1 wk. The reduction in serum bilirubin levels, the short duration of action and minimal confounding effects suggest that ZnPP may be an effective chemopreventive agent for the treatment of severe neonatal jaundice. PMID- 8725268 TI - Enzyme replacement with recombinant beta-glucuronidase in murine mucopolysaccharidosis type VII: impact of therapy during the first six weeks of life on subsequent lysosomal storage, growth, and survival. AB - Treatment of mucopolysaccharidosis type VII (MPS VII) mice with recombinant mouse beta-glucuronidase injections has been shown to deliver enzyme to most tissues and to reduce lysosomal storage during the first 6 wk of life. Here we determine the effect of enzyme therapy limited to the first 6 wk of life on survival and growth and follow the subsequent accumulation of lysosomal storage after beta glucuronidase treatment is discontinued. MPS VII mice received 28,000 U of beta glucuronidase i.v. at weekly intervals from birth to 6 wk of life and were killed at intervals up to 1 y after the last injection. By 29 d after the last enzyme injection, lysosomal storage in bone was no different in amount than that seen in untreated MPS VII mice. By 85 d, the fixed tissue macrophage system, meninges, and brain glia had also accumulated storage comparable to that seen in untreated controls. One year after treatment, lysosomal storage was similar to that of untreated MPS VII mice in all sites except cortical neurons, where there was still a slight reduction. All treated mice that were not killed earlier, lived longer, were larger, and had milder facial and skeletal deformities than untreated MPS VII mice. These data show that enzyme replacement therapy in MPS VII mice during the first 6 wk of life improve survival and growth. After treatment is discontinued, storage accumulates slowly in the brain and more rapidly in the fixed tissue macrophage system. Whether therapy continued later in life can further improve survival and growth remains to be established. PMID- 8725269 TI - Immunotitration analysis of cytosolic acetoacetyl-coenzyme A thiolase activity in human fibroblasts. AB - There are five known thiolases in human fibroblasts, and all but mitochondrial trifunctional protein (TFP) have thiolase activity toward acetoacetyl-CoA (AACoA). We investigated the contribution of mitochondrial acetoacetyl-CoA thiolase (AACoAT) (T2), cytosolic AACoAT (CT), and mitochondrial 3-ketoacyl-CoA thiolase (T1) to the total AACoAT activity in control human fibroblasts. Immunotitration of AACoAT activity with antibodies against T2, CT, or T1 was carried out in control fibroblasts, with the following results. In the case of AACoAT activity in the absence of potassium ion, 26-38%, 40-47%, and 11-20% of the total activity derived from CT, T1, and T2, respectively. The residual 6-9% total activity was not immunotitrated when three antibodies were used in combination. Hence, the contribution of peroxisomal 3-ketoacyl-CoA thiolase to the total AACoAT activity in the absence of potassium ion was at least less than 6-9%. Because the normal range of total AACoAT activity is relatively wide, it is difficult to evaluate CT defects based on a decrease of total AACoAT activity. Immunotitration with anti-CT antibody in six control fibroblasts revealed that CT activity ranges between 1.3 and 2.4 nmol/min/mg of protein. Immunotitration proved to be an accurate method to evaluate CT activity. The two cell lines from patients with CT deficiency have become extinct. PMID- 8725270 TI - Ethylmalonic aciduria is associated with an amino acid variant of short chain acyl-coenzyme A dehydrogenase. AB - Ethylmalonic aciduria is a common biochemical finding in patients with inborn errors of short chain fatty acid beta-oxidation. The urinary excretion of ethylmalonic acid (EMA) may stem from decreased oxidation by short chain acyl-CoA dehydrogenase (SCAD) of butyryl-CoA, which is alternatively metabolized by propionyl-CoA carboxylase to EMA. We have recently detected a guanine to adenine polymorphism in the SCAD gene at position 625 in the SCAD cDNA, which changes glycine 209 to serine (G209S). The variant allele (A625) is present in homozygous and in heterozygous form in 7 and 34.8% of the general population, respectively. One hundred and thirty-five patients from Germany, Denmark, the Czech Republic, Spain, and the United States were selected for this study on the basis of abnormal EMA excretion ranging from 18 to 1185 mmol/mol of creatinine (controls < 18 mmol/mol of creatinine). Among them, we found a significant overrepresentation of the variant allele. Eighty-one patients (60%) were homozygous for the A625 allele, 40 (30%) were heterozygous, and only 14 (10%) harbored the wild-type allele (G625) in homozygous form. By overexpressing the wild-type and variant protein (G209S) in Escherichia coli and COS cells, we showed that the folding of the variant protein was slightly compromised in comparison to the wild-type and that the temperature stability of the tetrameric variant enzyme was lower than that of the wild type. Taken together, the over-representation and the biochemical studies indicate that the A625 allele confers susceptibility to the development of ethylmalonic aciduria. PMID- 8725271 TI - Cathepsin A deficiency in galactosialidosis: studies of patients and carriers in 16 families. AB - Deficiency of lysosomal protective protein/cathepsin A in humans is the primary cause of galactosialidosis, a lysosomal storage disease characterized by combined deficiency of beta-galactosidase and neuraminidase. We have investigated 20 galactosialidosis patients and nine of their obligate heterozygous parents. A group of 12 patients with the early infantile type of the disease exhibited practically complete absence of cathepsin A activity, whereas eight patients with either the late infantile or the juvenile/adult type had 2-5% residual activity. Highest levels (5%) were present in two patients with milder clinical manifestations and later onset of the disease. In most fibroblast strains, beta galactosidase activity was 10-15% of normal levels, whereas neuraminidase was reduced to less than 4%. Interestingly, a substantial residual activity (10%) of the latter enzyme was detected in the patient with the mildest phenotype and the highest cathepsin A activity. Heterozygous values for cathepsin A were reduced on average to half of normal levels. However, in two cell strains, the activity was far below control range, and in these cases, neuraminidase activity was severely depressed. Finally, we showed that cathepsin A had considerable activity in chorionic villi and amniocytes, but was deficient in amniocytes from a pregnancy with an affected fetus, indicating the relevance of cathepsin A assay for prenatal diagnosis of galactosialidosis. PMID- 8725272 TI - Bilirubin has widespread inhibitory effects on protein phosphorylation. AB - The neurotoxic effects of bilirubin may involve modulation of neuronal protein phosphorylation systems. Using in vitro phosphorylation assays and a variety of protein substrates and purified protein kinases, we have studied the mechanism of bilirubin-induced inhibition of protein phosphorylation. Bilirubin was found to inhibit cAMP-dependent, cGMP-dependent, Ca(2+)- calmodulin-dependent, and Ca(2+) phospholipid-dependent protein kinases, irrespective of substrate properties. Fifty percent inhibition occurred at bilirubin concentrations varying from 20 to 125 microM. Kinetic analysis, using the isolated catalytic subunit of cAMP dependent kinase and a synthetic peptide substrate derived from the protein phospholemman, indicated that bilirubin (50 microM) decreased the apparent Vmax of the reaction, irrespective of whether ATP or peptide levels were varied, without significantly altering the apparent K(m) value. Thus our results indicate that bilirubin can inhibit catalytic domain(s) of protein kinases by apparent noncompetitive mechanism(s), presumably by interacting with noncatalytic domains on the enzyme. Given the key role of protein phosphorylation in cellular regulation, the widespread inhibitory effect of bilirubin on protein kinases may contribute to bilirubin neurotoxicity. PMID- 8725273 TI - Effect of acute feeding of diets of varying fatty acid composition on intestinal apolipoprotein expression in the newborn swine. AB - The purpose of this study was to determine the effects of dietary fatty acids of varying chain lengths and degrees of saturation on intestinal apolipoprotein (apo) B and A-I expression in the newborn piglet. Two-day-old female piglets received one of three isocaloric formulas containing 48% of total calories (120 kcal/kg/24 h) as medium-chain triglycerides (MCT) from MCT oil, intermediate chain saturated triglycerides (ICST) from coconut oil, or long-chain polyunsaturated triglycerides (LCPUT) from safflower oil by continuous duodenal infusion for 24 h. After in situ radiolabeling, jejunal and ileal mucosal apo B 48 and A-I were immunoprecipitated, and synthesis was expressed as percentage of total protein synthesis. Mucosal apo B and A-I mass was measured by ELISA as nanograms of apoprotein/microgram of total protein. Fifty percent less apo B jejunal synthesis was present in the ICST group versus the MCT and LCPUT groups (0.67 +/- 0.07, 1.19 +/- 0.20, and 1.25 +/- 0.15, respectively, mean +/- SEM, p < 0.05). Jejunal apo B mass was lower in the MCT group versus the ICST and LCPUT groups (0.10 +/- 0.02, 0.21 +/- 0.03, and 0.16 +/- 0.03, respectively, p < 0.05). Ileal apo B synthesis was lowest in the ICST group. No differences were found in ileal apo B mass. Two-fold higher jejunal apo A-I synthesis was found in the LCPUT group versus the MCT and ICST groups (14.18 +/- 1.69, 7.56 +/- 2.63, and 6.36 +/- 0.58, respectively, p < 0.01). No differences were found for jejunal apo A-I mass. In the ileum, the only difference was a higher apo A-I mass in the LCPUT group (p < 0.05). We conclude that in the newborn piglet intestinal apo B and A-I expression is acutely and differentially regulated by dietary lipid varying in fatty acid chain length and saturation. The patterns of regulation are complex and vary among specific apolipoproteins and regions of the small intestine and include co- and posttranslational mechanisms. PMID- 8725274 TI - Serine and glycine metabolism in hepatocytes from mid gestation fetal lambs. AB - Using stable isotopes of serine, glycine, and glutamine, the metabolism of serine and glycine was investigated in primary hepatocytes from six mid-gestation fetal lambs (mean gestational age = 81 +/- 6 d, normal gestation = 145 d). Serine production was 6.84 +/- 1.22 mumol/24 h/mg of DNA and exceeded serine utilization (3.76 +/- 1.44 mumol/24 h/mg of DNA) with a resultant net increase in medium serine of 2.58 +/- 1.70 mumol/24 h/mg of DNA. In contrast, glycine production (6.84 +/- 1.16 mumol/24 h/mg of DNA) was less than glycine utilization (12.10 +/- 1.78 mumol/24 h/mg of DNA) with a net decline in medium glycine of -5.44 +/- 2.03 mumol/24 h/mg of DNA. Of the serine produced, 50.4 +/- 4.3% was derived from glycine via the action of serine hydroxymethyltransferase (SHMT) and the glycine cleavage enzyme complex (GCS). Increasing the medium serine concentration resulted in an increase in serine utilization and sparing of the utilization of other amino acids. Biosynthesis of glycine from serine accounts for only 18.1 +/- 5.6% of glycine production, and this percentage is not affected by changes in medium serine concentration. Using 2.5-[15N2]glutamine as the tracer, an estimated 18 +/- 7% of serine production was derived from transamination reactions. The specific activity of both cytosolic and mitochondrial SHMT was constant for the duration of the cultures. We conclude that, in mid-gestation fetal ovine hepatocytes, there is net production of serine (with glycine as the primary metabolic source of this serine biosynthesis) and net glycine utilization. These data suggest that flux through SHMT and GCS accounts for 50% of serine biosynthesis in mid-gestation fetal ovine hepatocytes. The sparing of the utilization of other amino acids by serine suggests that serine a conditionally essential amino acid for the mid-gestation fetal liver. PMID- 8725275 TI - Postnatal changes in water content and proton magnetic resonance relaxation times in newborn rabbit tissues. AB - In the present study, using proton nuclear magnetic resonance relaxation (H1 NMR) measurements, an attempt was made to quantitate water fractions with different mobility in the skin, skeletal muscle, and liver tissues obtained from New Zealand white rabbit pups. Serial studies were carried out at the postnatal age of 0-1, 24, 48, and 72 h in pups nursed with their mothers and suckling ad libitum (group I) and in those pups separated from their mothers and completely withheld from suckling (group II). Tissue water content (desiccation method) and T1 and T2 relaxation times (H1 NMR method) were measured. Free, loosely bound, and tightly bound water fractions were calculated by applying multicomponent fits of the T2 relaxation curves. It was demonstrated that skin water content and T1 and T2 relaxation times decreased with age (p < 0.01), the decrease in T2 proved to be more pronounced in group II than in group I (p < 0.05). Muscle and liver water, and T1 and T2 relaxation times did not change with age in the suckling pups. In response to with-holding fluid intake muscle water remained constant, liver water increased paradoxically (p < 0.05). T1 relaxation time showed no consistent change in either tissues, whereas T2 relaxation time decreased significantly (muscle, p < 0.01) or tended to decrease (liver, p < 0.06). Using biexponential analysis fast and slow components of T2 relaxation curve could be distinguished that corresponded to the bound and free water fractions. Bound water accounted for 42-47%, 50-57%, and 34-40% of total tissue water in the skin, skeletal muscle, and liver, respectively, regardless of age and fluid intake. Triexponential fits of the T2 relaxation curve made possible the further partition of tissue water into tightly bound (fast component), loosely bound (middle component), and free (slow component) water fractions. In all tissues studied, loosely bound fraction predominated (skin, 48-64%; muscle, 54-65%; liver, 45-63%), followed by the free (skin, 26-45%; muscle, 23-32%; liver, 20 25%) and the tightly bound water fraction (skin, 6-14%; muscle, 10-16%; liver, 14 33%). Postnatal age and fluid intake had no apparent influence on this pattern of distribution. It is concluded that the majority of neonatal tissue water is motion-constrained. The free, the loosely bound, and the tightly bound water fractions appear to be interrelated and dependent on age, fluid intake, the tissues studied, and their hydration. PMID- 8725277 TI - The measurement of peripheral venous oxyhemoglobin saturation in newborn infants by near infrared spectroscopy with venous occlusion. AB - A novel, noninvasive method for measuring peripheral venous oxygen saturation (Svo2) in newborn infants using near infrared spectroscopy (NIRS) with venous occlusion is described, along with a comparison with co-oximetry of peripheral venous blood. Peripheral Svo2 was measured in 16 newborn infants by NIRS with venous occlusion, and the results were compared with those obtained by co oximetry of blood from a forearm vein. There was a significant correlation between the two measurements (r = 0.85, p < 0.0001) and close agreement between the two techniques (mean difference (co-oximeter-NIRS) 6%, limits of agreement 5.1% to 17.1%). Three different methods for analysis of the NIRS data were examined to determine which period during the venous occlusion provided the most accurate measurement. The agreement was the same for all methods of analysis. PMID- 8725276 TI - Comparison of heliox and oxygen as washing gases for the nitrogen washout technique in preterm infants. AB - The nitrogen washout technique usually involves exposure of the patient to 100% oxygen for several minutes. This may be dangerous in preterm infants who are at risk of retinopathy of prematurity (ROP). We wanted to know whether heliox (79% He, 21% O2) can be used instead of oxygen when determining functional residual capacity (FRC). FRC measurements were made in 14 preterm infants [median (range) gestational age at birth 34 wk (27-37 wk), and at time of study 36 wk (33-40 wk)] who were breathing room air. FRC was measured using a computerized infant pulmonary function system, beginning in random order with either 100% O2 followed by heliox or vice versa. There was no systematic difference between the two methods with regard to lung volume measurements: mean (SD) FRC values, corrected for body weight, were 22.9 (7.1) mL/kg for O2 and 23.4 (7.0) mL/kg for heliox. We did not observe a systematic influence of the type of washing gas used (heliox or oxygen) on FRC in these infants. Our results suggest that the use of heliox instead of pure oxygen may be a suitable and safer alternative for FRC measurements with the nitrogen washout technique in preterm infants who are breathing low concentrations of inspired oxygen and are still at risk of ROP. PMID- 8725278 TI - In vitro effects of magnesium sulfate in isolated intrapulmonary and mesenteric arteries of piglets. AB - Magnesium sulfate (MgSO4) has been proposed to be an efficient treatment in persistent pulmonary hypertension of the newborn. We compared the ability of MgSO4 to inhibit the responses to several vasoconstrictors in isolated intrapulmonary and mesenteric arteries from 10-17-d-old piglets. MgSO4 (3-100 mM) produced a slight vasodilator effect in pulmonary arteries precontracted with the thromboxane A2 mimetic U46619 (10(-6) M), noradrenaline (10(-5) M), and KCl (80 mM) (15.1 +/- 3.7%; 20 +/- 3.33%; 10.4 +/- 0.9% at 100 mM MgSO4 respectively). In contrast, in mesenteric arteries MgSO4, produced a marked vasodilation (80.4 +/- 4.0%, 93.1 +/- 3.46%, and 87.5 +/- 1.93% at 100 mM MgSO4, respectively, p < 0.01 versus pulmonary arteries). The vasodilator effect of MgSO4 was endothelium independent and reversed by increasing the extracellular Ca2+ concentration. After incubation for 1 h of pulmonary arteries with three different MgSO4 concentrations (0, 1.2, and 4.8 mM) there were no differences in the contractile responses to U46619 nor in the vasodilator effects of acetylcholine or sodium nitroprusside. Rapid removal of Mg2+ from bath medium produced a transient vasodilation which was more marked in pulmonary than in mesenteric arteries and was greatly reduced by the removal of endothelium or by the nitric oxide synthase inhibitor L-NAME (10(-4) M). We conclude that MgSO4 is a poor vasodilator of pulmonary arteries in vitro and at physiologic concentrations appears to inhibit nitric oxide release from the pulmonary endothelium. Thus, the possible beneficial clinical effects of MgSO4 in persistent pulmonary hypertension of the newborn do not seem to be related to a direct effect on pulmonary vascular smooth muscle. PMID- 8725279 TI - Ventral neural cadherin, a novel cadherin expressed in a subset of neural tissues in the zebrafish embryo. AB - Cadherins are calcium-binding transmembrane glycoproteins that are important mediators of cell-cell association. Here we describe a novel member of this gene family, zebrafish ventral neural cadherin (VN-cad). Multiple VN-cad transcripts are first detectable by Northern blots at 60% epiboly. In the developing neural tube, VN-cad RNA is first found in the neuroectoderm, directly above the notochord, and later was localized to the neural keel. At the 20-somite stage, VN cad transcripts are confined to the ventral neural tube, otic vesicle, midbrain, and diencephelon. Transcription of VN-cad RNA continues in adult fish. The embryonic pattern of expression is not significantly disrupted in cyclops or no tail mutants, which lack the floor plate and notochord, respectively. Therefore, neither of these structures is absolutely required for VN-cad expression. The localized pattern of VN-cad expression suggests a possible role for this adhesion molecule in the initial formation and subsequent differentiation of the central nervous system. PMID- 8725280 TI - Wilms' tumor suppressor gene is involved in the development of disparate kidney forms: evidence from expression in the Xenopus pronephros. AB - The Wilms' tumor suppressor gene (WT1) is required for the formation of the mammalian metanephros, or adult kidney, and for the normal development of the mesonephros, the major mammalian embryonic kidney. In this report the isolation of a Xenopus gene closely related to the mammalian WT1 gene in both sequence and splicing pattern is described. Expression of this gene, xWT1, is restricted to the developing nephric system until late tadpole stages, which expression also begins to be observed in the heart. Within the nephric system, expression is observed in the dorsal portion of the splanchnic lateral plate in tailbud embryos, and in the glomus of early tadpoles. No expression is observed in the pronephric tubules or pronephric duct. The WT1 gene is therefore expressed in a similar temporal and spatial pattern in the vascularized portion of the amphibian pronephroi and in the mammalian metanephroi, arguing that it probably plays a similar crucial role in the morphogenesis of these very different kidney forms. The absence of expression in the developing pronephric tubules indicates that xWT1 is not required for the epithelialization of the tubular portion of the pronephros. PMID- 8725281 TI - Expression of alternatively spliced and canonical basic fibroblast growth factor mRNAs in the early embryo and developing heart. AB - Previous studies in this laboratory have revealed the presence of substantial deposits of basic fibroblast growth factor (bFGF; FGF-2) in the myocardium from the earliest stages of heart development (Parlow et al. [1991] Dev. Biol. 146:139 147) and that an autocrine supply of bFGF is required for myocardial cell proliferation (Sugi et al. [1993] Dev, Biol, 157:28-37). Recently, an alternatively spliced isoform of bFGF, termed alt-bFGF, was described during later stages of embryogenesis, after heart morphogenesis is complete (Borja et al. [1993] Dev. Biol. 157:110-118). Because the antibody and nucleic acid probes used in our previous studies would have recognized canonical as well as alt-bFGF proteins and mRNAs, we have examined the expression of alt-and canonical bFGF mRNAs at early stages of embryogenesis, during which the initial differentiative and morphogenetic phases of heart development occur (Hamburger-Hamilton stages 3 24). Reverse transcription/polymerase chain reaction (RT/PCR) analysis detected the presence of both alt-bFGF and bFGF mRNAs in whole embryos as early as stage 3 and in the developing heart from the time of its initial appearance at stage 9. The presence of alt-bFGF mRNA was corroborated by RNase protection analysis which, in assessing RNA from whole embryos, revealed increasing levels of alt bFGF mRNA between stages 5-18, suggesting that expression of alt-bFGF is developmentally regulated. Utilization of a probe that simultaneously protects segments of both alt- and canonical bFGF mRNAs indicated that alt-bFGF was the more abundant FGF isoform in the developing embryo until stage 24, when equivalent expression of each isoform was detected. Similar analysis revealed that alt-bFGF was the more abundant isoform in the embryonic heart, but that its relative expression was not decreased at stage 24. PMID- 8725282 TI - Alternative transcript of the chick alpha 2(I) collagen gene is transiently expressed during endochondral bone formation and during development of the central nervous system. AB - Endochondral bone formation is characterized by several transitions in the pattern of collagen gene expression, the best characterized of which occurs during chondrogenesis. Prechondrogenic mesenchymal cells synthesize predominantly type I collagen; during chondrogenesis, type I collagen synthesis ceases and production of cartilage-characteristic collagens is initiated. We previously identified the molecular mechanism that mediates cessation of alpha 2(I) collagen synthesis in chondrocytes (Bennett and Adams [1990] J. Biol. Chem. 265:2223 2230). This mechanism involves a change in the transcription initiation site, resulting in an alternative transcript that cannot encode alpha 2(I) collagen. In this report we demonstrate that the alternative transcript appears only transiently in cartilage. Its initial appearance is coincident with the onset of high levels of type II collagen synthesis in differentiated chondrocytes. However, it disappears in hypertrophic cartilage, and production of the authentic alpha 2(I) collagen mRNA is reinitiated, contributing to synthesis of a high level of type I collagen in hypertrophic chondrocytes at the chondro-osseous junction. We also show that the alternative transcript is not restricted to cartilage during embryonic development, since it initially appears in presomite embryos, well before the appearance of cartilage. At early stages of embryo genesis the alternative transcript is restricted to tissues derived from neuroectoderm; its appearance in those tissues is also transient. These data suggest that production of the alternative transcript of the alpha 2(I) collagen gene may be required for cessation of alpha 2(I) collagen synthesis during chondrogenesis, but the alternative transcript may be involved in other important developmental programs as well. PMID- 8725283 TI - Understanding mammary gland development through the imbalanced expression of growth regulators. AB - Functional differentiation of mammary tissue progresses in distinct phases spanning puberty and pregnancy. Here we have analyzed and compared the effects of transforming growth factor beta 1 (TGF beta 1), TGF alpha, and whey acidic protein (WAP), the Notch-related cell fate protein Int3, and p53 and pRb on mammary development. We chose transgene expression from the WAP gene promoter which is only active in mammary alveolar cells. The imbalanced expression of these molecules specifically altered development and differentiation of the gland. While TGF alpha did not disturb alveolar outgrowth, little or no alveolar structures developed in the presence of Int3. TGF beta 1, WAP, and the expression of SV40 T-antigen-which inactivates p53 and PRb-reduced overall alveolar development. The expression of individual milk protein genes was affected differentially by the transgenes. A WAP-lacZ transgene served as an additional indicator of terminal differentiation of alveolar cells, Homogeneous expression of lacZ was seen in mice transgenic for lacZ, or for TGF alpha and lacZ. In contrast, only a few differentiated cells were observed in the presence of TGF beta 1 and Tag. Thus, the expression of growth regulators in the same defined subset of mammary cells results in distinct developmental changes and a specific pattern of alveolar differentiation. PMID- 8725284 TI - Differential expression of integrin alpha subunits supports distinct roles during lung branching morphogenesis. AB - Epithelial branching morphogenesis is a process by which a continuous epithelium, embedded in mesenchyme, forms tubules that extend and branch into the surrounding mesenchyme. The morphogenetic process is responsible for the architecture of many organs including the lung. Proper expression and function of extracellular matrix (ECM) molecules, such as collagens and laminins, are necessary for branching to occur normally. However, little is known about the role of epithelial cell surface molecules that mediate epithelial-matrix interactions during this process. We have studied the expression patterns of cell surface collagen and laminin integrin receptor alpha subunits, alpha 1, alpha 2, alpha 3, and alpha 6, in relation to that of collagen and laminin during lung branching morphogenesis. The alpha 1 integrin subunit was present on endothelia and smooth muscles around airways and large blood vessels. The mesenchyme expressed high levels of alpha 2 and alpha 6 but not alpha 3, whereas the epithelium expressed all three integrin subunits. In contrast to the widespread epithelial expression of alpha 3 and alpha 6, the epithelial expression of alpha 2 was restricted to branch tips. By performing in situ hybridization and immunofluorescence on serial sections, we found that alpha 2 protein expression on the epithelium correlated spatially and temporally with high level expression of collagen IV and laminin-1 mRNAs, suggesting that the alpha 2-expressing epithelial cells were in the process of producing and assembling their collagen and laminin matrices. While the expression of alpha 3 and alpha 6 on all lung epithelia suggests that these integrins may be important to lung epithelial development, the unique expression pattern of the alpha 2 subunit suggests that the alpha 2 beta 1 integrin may be important at branch tips either in the process of collagen/laminin synthesis and assembly or extension of the epithelial tubules into the mesenchyme. PMID- 8725285 TI - Developmental Modulation of a beta myosin heavy chain promoter-driven transgene. AB - The molecular mechanisms underlying heart and skeletal muscle-specific gene expression during development and in response to physioloic stimuli are largely unknown. Using a novel immunohistochemical procedure to detect chloramphenicol acetyltransferase (CAT), we have investigated, in vivo at high resolution, the ability of cis-acting DNA sequences within the 5' flanking region of the mouse beta myosin heavy chain (MyHC) gene (beta-MyHC) to direct appropriate gene expression throughout development. A 5.6-kb fragment 5' to the beta-MyHC's transcriptional start site was linked to the reporter gene encoding CAT (cat) and used to generate transgenic mice. The anti-CAT in situ assay described in this report allowed us to define the ability of the promoter fragment to direct appropriate temporal, tissue- and muscle fiber type-specific gene expression throughout early development. In skeletal muscles, the transgene expression profile mimics the endogenous beta-myHC's at all developmental stages and is appropriately restricted to slow (type I) skeletal fibers in the adult. Surprisingly, transgene expression was detected in both the atria and ventricles during embryonic and fetal development, indicating that ventricular specification involves elements outside the 5.6-kb fragment. In contrast, in the adult, hypothyroid conditions led to transgene induction specifically in the ventricles, suggesting that distinct regulatory mechanisms control fetal versus adult beta MyHC expression in the cardiac compartment. PMID- 8725286 TI - Fibroblast growth factor-2 stimulates embryonic cardiac mesenchymal cell proliferation. AB - The proliferation response of stage 36 chick atrioventricular valve mesenchymal cells to fibroblast growth factor-2 (FGF-2) was studied in the tissue-like environment of three-dimensional cell aggregates maintained in organ culture. The mitogenic effects of FGF-2 on mesenchymal tissue depended on the FGF-2-stimulated formation of a fibronectin-containing extracellular matrix. The matrix was absent in unstimulated aggregates, and co-localized with regions of actively proliferating cells in stimulated aggregates. Inhibition of fibronectin matrix formation by the inclusion of Arg-Gly-Asp-containing peptides, which compete with fibronectin for binding to the cell surface alpha 5 beta 1 integrin receptors, abolished the proliferation effects of FGF-2. Inhibition of sulfation of cell surface glycosaminoglycans by treatment with sodium chlorate significantly reduced both the formation of the fibronectin matrix and cell proliferation in response to FGF-2, suggesting an involvement of the low-affinity sulfated glycosaminoglycan FGF receptor system. Thus, the FGF-stimulated growth of embryonic atrioventricular valve mesenchyme in vitro involves the production of a fibronectin matrix. We suggest that the stimulation of the fibronectin matrix represents an essential element in growth factor signaling of mesenchymal tissue, with the matrix serving as an anchorage substratum for the proliferating cells. PMID- 8725287 TI - Regulated expression of cadherin-11 in human epithelial cells: a role for cadherin-11 in trophoblast-endometrium interactions? AB - Cadherin-11 is a novel member of the cadherin supergene family. Cadherin-11 expression is localized to mesenchymal tissue and specific regions of the neural tube during mouse embryogenesis. Here we report that cadherin-11 is spatiotemporally expressed in the epithelial cells of the human placenta. Cadherin-11 mRNA levels were low in freshly isolated cytotrophoblast cells but increased as the cytotrophoblast cells aggregated and fused to form syncytiotrophoblast cells in vitro. The increase in cadherin-11 mRNA levels was concomitant with a decrease in E-cadherin expression. Cadherin-11 was localized to the syncytial trophoblast and extravillous cytotrophoblasts, but not the villous cytotrophoblasts of the human placenta by immunohistochemistry. As both of the former cell types have intimate interactions with the endometrium, we examined cadherin-11 expression in the human endometrium. Cadherin-11 was detected in the glandular and surface epithelium of the endometrium at all stages of the menstrual cycle. However, cadherin-11 was abundant only in the stroma in the late secretory stage of the menstrual cycle. The accumulation of cadherin-11 in the stroma correlated with decidualization. Taken together, our observations demonstrate that cadherin-11 is expressed in certain epithelial cell lineages and suggest the possibility that cadherin-11 plays an important role in mediating trophoblast-endometrium interactions. PMID- 8725288 TI - Apoptosis removes chick embryo tail gut and remnant of the primitive streak. AB - Removal of transient features in morphogenesis of chick embryo tail is by programmed cell death. We used ApopTagTM (Oncor, Gaithersburg, MD) with the peroxidase/diaminobenzidine (DAB) procedure to correlate apoptosis with earlier reports of patterns of cell death in stage HH17-25 embryos, and our results suggest that the cell death inferred with supravital staining and appearance of cells in morphogenesis of the tail bud is programmed cell death called apoptosis. Apoptosis markers in tail bud are most abundant in the median cell cord of occluded degenerating tail gut. Tail bud mesenchyme marks for apoptosis most frequently in the ventrum of older stages, where cell death has been reported. Cells of the remnant of the primitive streak (Hensen's node) mark for apoptosis, suggesting that programmed cell death is a stop signal for axial organization at the caudal terminus. Apoptosis markers in postmembrane cloacal endoderm anticipate the transient cloacal fenestra. Lack of apoptosis markers in neural tube, notochord, and somites supports the suggestion of Schoenwolf ([1981] Anat, Embryol. (Beri.) 162:183-197) that cells of those areas in the tail bud are assimilated into the growing rump of the chick embryo. Lack of markers in neural tube of tail bud formed by secondary neurulation suggests that apoptosis is not involved in cavitation of medullary cord, but further investigation is necessary. A limited investigation of pharyngeal membranes and midgut, where cell death has not been reported to be as important in morphogenesis, did not show apoptosis markers in those tissues (Miller and Briglin [1994] "Cell Death in Development and Cancer," Houston: University of Texas MD Anderson Cancer Center, pp, 82-83). Absence of apoptosis markers in roof of gut tube suggests that the lower frequency of thymidine labeling reported for those cells (Miller [1986] Anat. Rec. 214: 87A) is not a result of apoptosis. Clearly marked cells correlated with expected locations of migrating neural crest and primordial germ cells in these stages, but distribution of apoptosis markers was not abundant or general for either cell type. PMID- 8725289 TI - Fibronectin mRNA alternative splicing is temporally and spatially regulated during chondrogenesis in vivo and in vitro. AB - Fibronectin, a component of the extracellular matrix in a variety of tissues, participates in many critical cellular processes, including differentiation, adhesion, and migration. A positive correlation exists between the presence of fibronectin and the onset of chondrogenesis, the differentiation of mesenchyme into cartilage. Heterogeneity in the structure of fibronectin is largely due to the alternative splicing of at least three exons (IIIB, IIIA, and V) during processing of a single primary transcript. We have previously shown that the fibronectin mRNA splicing patterns change during chondrogenesis (Bennett et al. [1991] J. Biol. Chem, 266:5918-5924). All of the fibronectin mRNAs from prechondrogenic chick limb mesenchyme contain exons IIIB, IIIA, and V (B + A + V +), whereas all of the fibronectin mRNAs from chick cartilage contain exons IIIB and V but do not contain exon IIIA (B + A - V +). In this study, we show that fibronectin mRNAs containing exon IIIA (FN-A) and/or the mRNAs containing exon IIIB (FN-B) are expressed in a specific and different spatiotemporal manner in the developing chick limb in vivo, as well as in limb mesenchymal cells undergoing chondrogenesis in vitro. Specifically, in situ hybridization reveals that FN-B mRNAs are present throughout the various stages (HH 20-30) of limb cartilage development in vivo, whereas FN-A mRNAs disappear following the condensation phase of chondrogenesis and absent from the resulting cartilage, Chick limb cartilage fibronectin mRNAs are therefore B + A-, as in other embryonic cartilage tissues. Furthermore, limb mesenchymal cells undergoing chondrogenesis in vitro lose FN-A mRNAs immediately following condensation, recapitulating the events that occur during chondrogenesis in vivo. These results suggest an important role for fibronectin mRNA alternative splicing during chondrogenic differentiation. PMID- 8725290 TI - Development of olfactory glomeruli: temporal and spatial interactions between olfactory receptor axons and mitral cells in opossums and rats. AB - Mitral cells are the primary output neurons of the vertebrate olfactory bulb and are major recipients of sensory input from the periphery. The morphogenesis of mitral cell dendrites was followed to elucidate their early spatial and temporal interactions with olfactory receptor neurons and glia during the construction of olfactory glomeruli. Monodelphis domestica, a marsupial born at an extremely immature stage, and rats were examined. Mitral cells were retrogradely labeled by application of the lipophilic dye 1,1' dihexadecyl-3,3,3'3'-tetramethylin docarbocyanine perchlorate (DiI) to the lateral olfactory tract. In double labeling experiments, olfactory receptor neurons were stained with 3,3' dihexadecyloxacarbocyanine perchlorate (DiO), or olfactory nerve Schwann cells were visualized using S-100 protein immunohistochemistry. Tissue was examined with a confocal laser scanning microscope. Some preparations were subsequently investigated with an electron microscope. In Monodelphis, differentiation of mitral cells starts with an outgrowth of numerous, uniform, and widespread dendrites. As soon as terminals of olfactory receptor axons coalesce into glomerular knots within the presumptive glomerular layer, dendrites of individual mitral cells innervate several adjacent glomeruli where they receive sensory synaptic input. With maturation, supernumerary mitral cell dendrites retract, leaving one primary dendrite bearing a terminal glomerular tuft. Simultaneously, secondary dendrites begin to arise. The formation of glomeruli begins earlier and progresses faster in the rat compared to Monodelphis. Nevertheless, mitral cell differentiation in both species follows a common sequence: overproduction of dendrites, selection of usually one primary apical dendrite, and elimination of supernumerary processes. Since olfactory receptor neurons form synaptic contacts with the widespread mitral cell dendrites, considerable synaptic rearrangement must occur within the olfactory glomeruli during maturation. PMID- 8725291 TI - Colocalization of ionotropic glutamate receptor subunits with NADPH-diaphorase containing neurons in the rat mesopontine tegmentum. AB - Tegmental cholinergic neurons vary their discharge patterns across the sleep-wake cycle, and glutamate is suggested to play an important role in determining these firing patterns. Cholinergic and noncholinergic neurons in the mesopontine tegmentum have different susceptibilities to various excitotoxins, presumably because of heterogeneity in the expression of glutamate receptor subtypes in this area. By using a double-labeling procedure that combines nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-diaphorase) histochemistry and avidin biotin-peroxidase immunocytochemistry with diaminobenzidine as the chromogen, we compared the colocalization of AMPA receptor subunits GluR1, GluR2/3, and GluR4, kainate receptor subunits GluR5/6/7, and an NMDA receptor subunit NMDAR1 on NADPH diaphorase-positive (cholinergic) neurons in the mesopontine tegmentum. Throughout the brainstem, neurons immunoreactive for GluR2/3 and NMDAR1 were most numerous, whereas neurons labeled for GluR1, GluR4, and GluR5/6/7 were less common. Specifically within the mesopontine tegmentum, the proportion of double labeled neurons in the diaphorase-containing cell population was highest with GluR1 (43%) and lowest with GluR5/6/7 (12%). Regardless of the receptor subunit type, the greatest numbers of double-labeled neurons were observed in the pedunculopontine tegmental nucleus pars compacta and the fewest in the dorsal aspect of the laterodorsal tegmental nucleus. In addition, there were regional differences in the relative expression of receptor subunits and diaphorase positive neurons across the subdivisions of the tegmental cholinergic column. Because each ionotropic subunit confers distinctive properties to a receptor channel, the present results suggest that mesopontine cholinergic neurons have nonuniform responses to glutamate and are also discriminable from basal forebrain cholinergic neurons in terms of glutamate receptor configuration. PMID- 8725292 TI - Target-specific differences in the dendritic morphology and neuropeptide content of neurons in the rat SCG during development and aging. AB - Our purpose in this work was to investigate the role of target tissues in the regulation of dendritic morphology from sympathetic neurons during development and aging. Neurons were retrogradely labeled from three targets, the iris, the submandibular gland (SMG), and the middle cerebral artery (MCA). They were then fixed and intracellularly injected to demonstrate their dendritic arborizations. Dendritic geometry varied quantitatively in sympathetic neurons innervating different target tissues at all stages of development. Neurons innervating the iris had the largest cell bodies and most extensive dendritic arborizations, whereas the vasomotor neurons were the smallest. The number of primary dendrites, however, did not vary significantly between the different neuronal populations. The growth of dendritic arborizations during postnatal development and their atrophy in old age were not concordant in the different neuron populations we studied. Neurons innervating the MCA and the iris ceased dendritic growth early in postnatal development, whereas the dendritic complexity of neurons supplying the SMG increased well into adulthood. By contrast, dendritic atrophy was seen in aged MCA- and SMG-projecting neurons but not in those innervating the iris, suggesting, with other evidence, correlated and distinct patterns of growth and atrophy in axons and dendrites of mature sympathetic neurons projecting to different targets. Swollen dendrites and protuberances on cell soma were a prominent feature of aged neurons. In addition to the target-specific variation in neuronal morphology, we observed diversity in neurotransmitter phenotype. For example, neuropeptide Y was expressed in iridial but not SMG-projecting neurons. These results show a range of age- and target-specific differences in the dendritic morphology and neuropeptide content of sympathetic neurons that may be a result of differing trophic interactions with their target tissues. PMID- 8725293 TI - Subnuclear localization of FOS-like immunoreactivity in the rat parabrachial nucleus after nociceptive stimulation. AB - The effect of noxious stimulation on the expression of FOS-like immunoreactivity (FOS-LI) in neurons of the parabrachial nucleus (PB) was studied in awake, freely moving rats. In one series of experiments, the rats were subjected to noxious mechanical stimulation (pinch) of either the nape of the neck or the base of the tail for 20 seconds every 5 minutes for 90 minutes, and then they were killed by transcardial perfusion after 45-210 minutes. Control animals received innocuous mechanical stimulation (brush) of the tail. Noxious stimuli resulted in FOS-LI in neurons in the dorsal part of the lateral PB, with heavy labeling in the superior lateral (PBsl) and the dorsal lateral (PBdl) subnuclei. FOS-LI was also elicited in the central lateral subnucleus (PBcl) and, although much more sparsely, in the external lateral subnucleus and the Kolliker-Fuse nucleus. Tail and neck stimulation resulted in similar labeling patterns, but more neurons, particularly in PBsl, expressed FOS-LI after pinch of the tail than of the neck. In another series of experiments, rats received injection of 5% formalin into one hindpaw. After 75-90 minutes, FOS-LI was seen in the same parts of PB as after noxious mechanical stimulation. The heaviest labeling was seen on the side contralateral to the injection side, with statistically significant (P < 0.05) side differences present in PBsl and PBdl. In a third series of experiments, rats were hemisected at low cervical-upper thoracic segments, allowed 2 weeks to recover, and then given formalin injections in both hindpaws. Significantly more neurons were FOS labeled in PBdl, PBsl, and PBcl on the side contralateral to the hemisection than on the ipsilateral side. These observations are discussed in relation to the organization of the spinal afferent input and the efferent connections of PB. It is concluded that the FOS-LI expression in PBdl and PBsl and probably also in PBcl, to a large extent, is evoked by the ascending spinal nociceptive input to PB. Because these subnuclei project to several hypothalamic regions, it is suggested that neurons in PB that express FOS after noxious mechanical and chemical stimulation primarily are involved in autonomic and homeostatic responses to behavioral situations that involve tissue-damaging stimuli. PMID- 8725294 TI - Two types of corticopulvinar terminations: round (type 2) and elongate (type 1). AB - Corticopulvinar axons were anterogradely labeled by Phaseolus vulgaris leucoagglutinin injections in the occipitotemporal cortex of the macaque to determine quantitative parameters of divergence and convergence, arbor size and shape, and distribution of terminal specializations. Forty individual axons were analyzed by serial section reconstruction and divided into two major groups. The majority of axons have numerous (typically 500-1,000) small, spinous endings (boutons terminaux). These axons have terminal fields that are beam-like or elongated (E, corresponding to classical type 1) and highly divergent (1.0-3.0 mm). These frequently innervate several of the traditionally designated pulvinar subdivisions; namely inferior pulvinar (PI) and the ventral part of interal pulvinar (PL); medial pulvinar (PM) and dorsal PL, and (one axon) PM, dorsal PL, and PI. Some axons, however (R or round, corresponding to classical type 2), have a small number (typically 70-160) of primarily large, beaded endings (boutons en passant), which concentrate in sharply delimited, round arbors (diameters 100-125 microns). R axons appear to be larger caliber than E axons (1.0-1.5 microns vs. 0.5-1.0 micron, respectively). These differences in phenotype are probably associated with distinct types of projection neurons. In visual areas, corticopulvinar terminations are reported to originate from pyramidal cell subpopulations in layer 5. Indirect evidence, presented here, suggests that the more numerous medium-sized neurons give rise to E axons, and the sparser giant pyramids give rise to R corticopulvinar axons. If this is correct, corticopulvinar connectivity may be involved in multiple transformations. Spatially, axons of giant neurons (with basal dendrites that collect intracortically from a disc-like area, about 1.0 mm in diameter) converge onto a small number of pulvinar neurons. Axons of medium neurons (with basal dendrites that occupy a small intracortical disc, about 0.3 mm in diameter) diverge over 1.0-3.0 mm in the pulvinar and may form many contacts. Giant neurons, although numerically few in relation to medium pyramids (1 or 2: 50?), are likely to have distinctive membrane properties (functionally equivalent to bursting neurons?). Their larger boutons and axon caliber may be associated with a faster transmission that compensates for their small numbers. In primates, the E and R duality does not characterize cortical projections to the caudate, lateral geniculate nucleus, pons, or superior colliculus and thus may be essentially linked to pulvinar-specific processes. PMID- 8725295 TI - Fos expression in forebrain afferents to the hypothalamic paraventricular nucleus following swim stress. AB - The paraventricular nucleus of the hypothalamus (PVN) serves as the origin of the final common pathway in the secretion of glucocorticoid hormones in response to stress. Various stress-related inputs converge upon the cells of the medial parvocellular division of the PVN. These neurons, which synthesize and release corticotropin-releasing hormone, arginine vasopressin, and other secretagogues, are responsible for a cascade of events which culminates in the adrenocorticotropin-induced release of corticosteroids from the adrenal cortex. Previous data have suggested complex afferent regulation of PVN neurons, although the neuronal pathways by which the effects of stress are mediated remain to be fully disclosed. The present experiment sought to identify forebrain areas potentially involved in afferent regulation of the PVN in response to an acute stressor. Discrete injections of the retrograde tracer Fluoro-gold were delivered to the PVN, and rats were subsequently subjected to an acute swim stress. Brains were processed immunocytochemically for the simultaneous detection of the tracer and Fos, the protein product of the immediate early gene c-fos, utilized as a marker for neuronal activation. The majority of Fluoro-gold/Fos labeled neurons were detected in the parastrial nucleus, the medial preoptic area, the anterior hypothalamic area, the dorsomedial hypothalamic nucleus and adjacent posterior hypothalamic area, and, to a lesser extent, the supramammillary nucleus. These findings are discussed in relation to neural pathways mediating activation and inhibition of the hypothalamic-pituitary-adrenocortical axis. PMID- 8725296 TI - Organization of ventrolateral periolivary cells of the cat superior olive as revealed by PEP-19 immunocytochemistry and Nissl stain. AB - Ventrolateral periolivary cell groups, through their descending projections to the cochlear nucleus (CN) and local projections to principal nuclei of the superior olive, may participate in brainstem mechanisms mediating such tasks as signal detection in noisy environments and sound localization. Understanding the function of these cell groups can be improved by increased knowledge of the organization of their synaptic inputs in relation to their cellular characteristics. Immunocytochemistry for PEP-19 (a putative calcium binding protein) reveals four patterns of immunolabeling within the ventrolateral periolivary region. Three of the patterns, which have distinct fiber and punctate labeling characteristics, help to define three subdivisions of the lateral nucleus of the trapezoid body (LNTB). The fourth pattern defines two other nuclei, the anterolateral periolivary nucleus (rostral) and the posterior periolivary nucleus (caudal), which display many immunoreactive cell bodies but little fiber and punctate labeling. One of the subdivisions of the LNTB contains large PEP-19 immunolabeled puncta arranged in pericellular nests. Analysis of Nissl-stained sections reveals a neuronal population that resembles globular cells of the ventral cochlear nucleus (VCN) and which colocalizes with pericellular nests of large immunolabeled puncta. Cell counts reveal that roughly 10,000 neurons constitute the cat ventrolateral periolivary region, 9,000 of which are found in the LNTB. Three-dimensional reconstructions of auditory brainstem nuclei clarify the complex spatial relationships among these structures. PMID- 8725297 TI - Somatotopic organization of the facial lobe of the sea catfish Arius felis studied by transganglionic transport of horseradish peroxidase. AB - To reveal the somatotopical organization of the facial lobe (FL), a primary medullary gustatory nucleus in the sea catfish Arius felis, the central projections of the peripheral rami of the facial nerve innervating taste buds located across the entire body surface and rostral oral regions were traced by means of horseradish peroxidase neurohistochemistry. The maxillary barbel, lateral mandibular barbel, medial mandibular barbel, and trunk-tail branches project to four different longitudinal columns (i.e., lobules) extending rostrocaudally in the FL. The trunk-tail lobule, which is located dorsolateral to the barbel lobules, lies in the anterior two-thirds of the FL. The tail is represented in a more rostral portion of the trunk-tail lobule than the trunk, indicating that the rostrocaudal trunk axis is represented in the trunk-tail lobule in a posteroanterior axis. The pectoral fin branch ends in an intermediate region of the FL, whereas the hyomandibular, ophthalmic, lower lip, upper lip, and palatine branches terminate in discrete regions of the caudal one-third of the FL. These results reveal a sharply defined somatotopical organization of the FL of Arius and support the hypothesis that the number and lengths of the barbel lobules within the FL of catfishes are directly related to the number and relative lengths of the barbels. An additional subcolumn, the intermediate nucleus of the FL (NIF), which develops in the medioventral region of the caudal two-thirds of the FL, receives projections in a diffuse somatotopical fashion from the barbels, lower lip, and palatine branches. Trigeminal fibers of the barbel and lower lip branches project in a somatotopic fashion to the FL. The present findings suggest that the FL of Arius is highly organized somatotopically to detect, by tropotaxis, precise spatial information concerning taste and tactile stimuli in the environment. PMID- 8725298 TI - Central projections and somatotopic organisation of trigeminal primary afferents in pigeon (Columba livia). AB - Injections of cholera toxin B-chain conjugated to horseradish peroxidase into individual peripheral branches of the trigeminal nerve or into the trigeminal ganglion showed that an ascending trigeminal tract (TTA) terminated in distinct ventral and dorsal divisions of the principal sensory nucleus (PrVv and PrVd, respectively), and a descending tract (TTD) terminated within pars oralis, pars interpolaris, and pars caudalis divisions of the nucleus of TTD (nTTD) and within the dorsal horn of the first six cervical spinal segments. In PrVd, mandibular, ophthalmic, and maxillary projections were predominantly located dorsally, ventrally, and medially, respectively. In nTTD, mandibular projections lay dorsomedially, ophthalmic projections lay ventrolaterally, and maxillary projections lay in between. At caudal medullary and spinal levels, mandibular projections were situated medially, ophthalmic projections were situated laterally, and maxillary projections were situated centrally. The terminations within the dorsal horn were most dense in laminae III and IV and were least dense in lamina II, with laminae III-IV also receiving topographically organised contralateral projections. Extratrigeminal projections were mainly to the external cuneate nucleus by way of a lateral descending trigeminal tract (lTTD; Dubbeldam and Karten [1978] J. Comp. Neurol. 180:661-678) and to the region of the tract of Lissauer and lamina I of the dorsal horn. Other projections were to a region medial to the apex of pars interpolaris, to the nuclei ventrolateralis anterior (Vla) and presulcalis anterior (Pas) of the solitary complex, and sparsely to the lateral reticular formation (plexus of Horsley) ventral to TTD. No projections were seen to the trigeminal motor nuclei or to the cerebellum. PMID- 8725299 TI - Proctolinlike immunoreactivity and identified neurosecretory cells as putative substrates for modulation of courtship display behavior in the blue crab, Callinectes sapidus. AB - Pheromonally stimulated courtship display (CD) behavior in male blue crabs (Callinectes sapidus) is characterized by rhythmic waving of the fifth legs. The waving of the fifth legs is modulated by proctolin in freely moving crabs and in reduced preparations. To begin to identify an anatomical substrate for CD behavior we have localized putative proctolinergic cells and described the morphology of neurosecretory neurons known to oscillate during pheromonal stimulation of reduced preparations. Proctolin-induced CD occurs with developmental and seasonal dependence. Male crabs altered hormonally by eyestalk ligation spontaneously produce CD behavior. We have localized proctolinlike immunoreactivity (PIR) in the central nervous system (CNS) and compared this immunoreactivity across sexes, developmental stage, eyestalk ligation, and seasonal conditions to determine whether or not expression of PIR is correlated with CD behavior. PIR was found in most areas of the CNS. Clusters of PIR positive cells were found in the sinus gland and eyestalk ganglia, olfactory neuropil with associated cell bodies, and in a large cell cluster in the subesophageal region of the ventral nerve cord. Three pairs of cell bodies in different cell body groups in the brains of adult crabs stained positively for PIR but did not stain in the youngest juvenile animals. Comparison of PIR distribution with toluidine blue studies of the ventral nerve cord indicated a high likelihood that cells in the PIR-positive cluster of the subesophageal ganglia were also members of the cluster of neurons identified as oscillatory neurosecretory neurons. PMID- 8725300 TI - Postnatal changes in NMDAR1 subunit expression in the rat trigeminal pathway to barrel field cortex. AB - N-methyl-D-aspartate (NMDA) type glutamate receptors are constituted of one obligatory subunit (NR1), expressed as eight splice variants, combined with one or more of four NMDAR2 subunits. Polyclonal antibodies were produced to an N terminal domain of the NR1 subunit that recognize all eight splice variants. The antibody was used to localize NR1 in the trigeminal pathway to barrel field cortex in rats. The distribution and density of NR1 changes between birth (postnatal day 0 = P-0) and P-360. The trigeminal nuclei already contain a high level of NR1 immunoreactivity on the day of birth. The ventral posterior lateral, ventral posterior medial, and posterior nucleus, medial division, thalamic nuclei show fluctuations in NR1 immunoreactivity levels, starting at birth with moderate densities in neuropil which decrease at P-7, and peak again in neuronal cell bodies as well as the neuropil at P-21. In the cortex, the density of NR1 in layer VI fluctuates with low points at P-7 and P-40. Superficial cortical layers I, II, and III reach adult levels at P-14 and remain high. NR1 levels decrease sharply in layer IV just prior to P-40 and then slowly recover over the next 3 months to stabilize at moderate levels in the adult. In addition to neuronal expression there is a transient high level of labeling in glial cells with a peak density of staining at P-21. The results emphasize that NR1 subunit expression is finely regulated in rat somatic sensory pathways for periods as long as 7-8 weeks after birth in the barrel field cortex. PMID- 8725301 TI - Distribution of galanin-like immunoreactive elements in the brain of the adult lamprey Lampetra fluviatilis. AB - Galanin is a brain-gut peptide present in the central nervous system of vertebrates and invertebrates. The distribution of galanin-like immunoreactive perikarya and fibers in the brain of the river lamprey Lampetra fluviatilis (Agnatha) has been studied immunocytochemically by using antisera against rat and porcine galanin. Galanin-like immunoreactive perikarya were seen in the telencephalon and mediobasal diencephalon. In the telencephalon, they were present in the nucleus olfactorius anterior, nucleus basalis, and especially, in the nucleus commissurae anterioris. The diencephalon contained most of the immunoreactive neurons. They were located in the nucleus commissurae praeinfundibularis, nucleus ventralis hypothalami, nucleus commissurae postinfundibularis, nucleus ventralis thalami, and nucleus dorsalis thalami pars medius. Most of the galanin-like immunoreactive infundibular neurons showed apical processes contacting the cerebrospinal fluid. Immunoreactive fibers and terminals were widely distributed throughout the neuraxis. In the telencephalon, the richest galaninergic innervation was found in the nucleus olfactorius anterior, lobus subhippocampalis, corpus striatum, and around the nucleus septi and the nucleus praeopticus. In the diencephalon, the highest density of galanin like immunoreactive fibers was seen in the nucleus commissurae postopticae, nucleus commissurae praeinfundibularis, nucleus ventralis hypothalami, nucleus dorsalis hypothalami, and neurohypophysis. In the mesencephalon and rhombencephalon, the distribution of immunoreactive fibers was heterogeneous, being most pronounced in a region between the nucleus nervi oculomotorii and the nucleus interpeduncularis mesencephali, in the nucleus isthmi, and in the raphe region. A subependymal plexus of immunoreactive fibers was found throughout the ventricular system. The distribution of immunoreactive neurons and fibers was similar to that of teleosts but different to those of other vertebrate groups. The possible hypophysiotropic and neuroregulatory roles of galanin are discussed. PMID- 8725302 TI - Efferent connectivity of the hippocampal formation of the zebra finch (Taenopygia guttata): an anterograde pathway tracing study using Phaseolus vulgaris leucoagglutinin. AB - The avian hippocampal formation (HP) is considered to be homologous to the mammalian hippocampus, being involved in memory formation and spatial memory in particular. The subdivisions and boundaries of the pigeon hippocampus have been defined previously by various morphological methods to detect further similarities with the mammalian homologue. We studied the efferent projections of the zebra finch hippocampus by applying Phaseolus vulgaris leucoagglutinin, and three main subdivisions were distinguished on the basis of the connectivity patterns. Dorsolateral injections gave rise to projections innervating the rostralmost extension of the HP, a laminar complex including the dorsal and ventral hyperstriata and the lamina frontalis superior, the rostral lobus parolfactorius, the medial and ventral paleostriatal regions, the lateral septal nucleus, the nucleus of the diagonal band, the dorsolateral corticoid area, the archistriatum posterius, and the nucleus taeniae in the telencephalon. In the diencephalon, labelled axons were seen in the periventricular and lateral hypothalamus, including the lateral mammillary nuclei, and in the dorsolateral and the dorsomedial posterior thalamic nuclei, whereas, in the midbrain, only the area ventralis of Tsai contained hippocampal fibres. With the exception of the bilateral archistriatal efferents, all projections were ipsilateral. Dorsomedial injections gave rise to a local fibre system that was almost completely restricted to the ipsilateral hippocampal formation. In addition, lectin containing fibres continued in the dorsal septal region and a thin band in the hyperstriatum accessorium, adjacent to the lateral ventricle. Ventral injections gave rise to axons innervating ipsilaterally the dorsolateral subdivision, and bilaterally the medial septal nuclei and the contralateral ventral hippocampus. PMID- 8725303 TI - Comparison of cerebellothalamic and pallidothalamic projections in the monkey (Macaca fuscata): a double anterograde labeling study. AB - To address the question of segregated projections from the internal segment of the globus pallidus (GPi) and the cerebellar nuclei (Cb) to the thalamus in the monkey, we employed a double anterograde labeling strategy combining the anterograde transport of horseradish peroxidase conjugated to wheat germ agglutinin (WGA-HRP) with biotinylated dextran amine (BDA) transport. The tissue was processed sequentially for WGA-HRP, and then BDA immunohistochemistry using two different chromogens. Since the two labels were easily distinguishable on the same histological section, the interrelationship between the cerebellar and pallidal projection systems could be directly evaluated. We found that both the cerebellothalamic and pallidothalamic label consisted of dense plexuses of labeled fibers and swellings in a patch-like configuration. The patches or foci of labeling were distributed either as dense single label or as interdigitating patches of double label. We found dense single label in the central portion of the ventral anterior nucleus pars principalis (VApc) and the ventral lateral nucleus pars oralis (VLo) following the GPi injections or in the central portion of the ventral posterior lateral nucleus pars oralis (VPLo) and nucleus X (X) following the cerebellar nuclei injections. Complementary interdigitating patches of WGA-HRP and BDA labeling were found primarily in transitional border regions between thalamic nuclei. On occasion, we found overlap of both labels. We observed a gradient pattern in the density of the pallidothalamic and cerebellothalamic projections. The pallidothalamic territory included VApc, VLo, and the ventral lateral nucleus pars caudalis (VLc), with the density of these projections decreasing along an anterior to posterior gradient in the thalamus. Occasional patches of pallidal label were found in VPLo and nucleus X. Conversely, the density of cerebellothalamic projections increased along the same gradient, with the cerebellothalamic territory extending anteriorly beyond the cell-sparse zones of VPLo, X, and VLc to include VLo and VApc also. These data suggest that although the cerebellar and pallidal projections primarily occupy separate thalamic territories, individual thalamic nuclei receive differentially weighted inputs from these sources. PMID- 8725304 TI - Immunohistochemical localization of ORL-1 in the central nervous system of the rat. AB - A novel member of the opioid receptor family (ORL-1) has been cloned from a variety of vertebrates. ORL-1 does not bind any of the classical opioids, although a high affinity endogenous agonist with close homology to dynorphin has recently been identified. We have generated a monoclonal antibody to the N terminus of ORL-1 to map areas of receptor expression in rat central nervous system (CNS). Intense and specific immunolabeling was observed in multiple areas in the diencephalon, mesencephalon, pons/medulla, and spinal cord. In the telencephalon, intense labeling was observed in the neuropil throughout layers II V in the neocortex, the anterior olfactory nuclear complex, the pyriform cortex, the CA1-CA4 fields and dentate gyrus of the hippocampus, and in many of the septal and basal forebrain areas. In contrast to other members of the opioid receptor family, light labeling for ORL-1 was observed in telencephalic areas such as caudate-putamen. In the cerebellum, ORL-1 immunoreactivity was only observed in the deep nuclei. Throughout the CNS the majority of labelling was localized to fiber processes and fine puncta, although labeled scattered perikarya were observed in a few brain areas such as the hilus dentate in the hippocampus and some nuclei in the brainstem and spinal cord. The present mapping study is consistent with the reported distribution of ORL-1 mRNA and provides the first immunohistochemical report on anatomical and cellular distribution of ORL-1 receptor in the rat CNS. PMID- 8725305 TI - Anatomical basis for audio-vocal integration in echolocating horseshoe bats. AB - Neurophysiological recordings suggest that audio-vocal neurons located in the paralemniscal tegmentum of the midbrain in horseshoe bats provide an interface between the pathways for auditory sensory processing and those for the motor control of vocalization. To verify these physiological results anatomically, the projection pattern of the audio-vocally active area in the paralemniscal tegmentum was investigated by using extracellular tracer injections of wheat germ agglutinin conjugated to horseradish peroxidase. Several nuclei of the lemniscal auditory pathway (dorsal nucleus of the lateral lemniscus, central nucleus of the inferior colliculus, lateral superior olive) as well as the nucleus of the central acoustic tract appear to project to the paralemniscal tegmentum. Other possible sources of afferent projections are a small but distinctly labeled structure within the lateral hypothalamic area, the substantia nigra pars compacta, the deep mesencephalic nucleus, the rostral portion of the inferior colliculus, the deep and intermediate layers of the superior colliculus, and several small areas in the rhombencephalic reticular formation. No direct efferent projection from the audio-vocally active area of the paralemniscal tegmentum to primarily auditory structures was found. Instead, the main targets were structures that are involved in the control of different motor patterns. These targets include the deep and intermediate layers of the superior colliculus and the dorsomedial portion of the facial nucleus, both of which most probably control pinna movements in cats, and the reticular formation medial and caudal to the facial nucleus and rostral to the nucleus ambiguus, which represents an area involved in the control of vocalization. Hence, the anatomical projection pattern suggests that the paralemniscal tegmentum in horseshoe bats serves as a link between the processing of auditory information and the control of vocalization and related motor patterns. PMID- 8725306 TI - Characterization and ontogeny of synapse-associated proteins in the developing facial and hypoglossal motor nuclei of the Brazilian opossum. AB - The characterization and ontogeny of synapse-associated proteins in the developing facial and hypoglossal motor nuclei were examined in the Brazilian opossum (Monodelphis domestica). Immunohistochemical markers utilized in this study were the synaptic vesicle-associated proteins synaptophysin and synaptotagmin; a synaptic membrane protein, plasma membrane-associated protein of 25 kDa (SNAP-25); a growth cone protein, growth-associated phosphoprotein-43 (GAP 43); and the microtubule-associated proteins axonal marker tau and dendritic marker microtubule-associated protein-2 (MAP-2). In this study, we have found that, during the first 10 postnatal days (1-10 PN), the facial motor nucleus lacked immunoreactivity for synaptophysin, synaptotagmin, GAP-43, tau, and SNAP 25. After 10 PN, immunoreactivity increased in the facial motor nucleus for synaptophysin, synaptotagmin, GAP-43, and tau, whereas immunoreactivity for SNAP 25 was not evident until between 15 and 25 PN. Conversely, immunoreactivity for MAP-2, was present in the facial motor nucleus from the day of birth. In contrast, the hypoglossal motor nucleus displayed immunoreactivity from 1 PN for synaptophysin, synaptotagmin, SNAP-25, GAP-43, tau, and MAP-2. These results suggest that the facial motor nucleus of the opossum may not receive afferent innervation as defined by classical synaptic markers until 15 PN and, further, that characteristic mature synapses are not present until between 15 and 25 PN. These results indicate that there may be a delay in synaptogenesis in the facial motor nucleus compared to synaptogenetic events in the hypoglossal motor nucleus. Because the facial motor nucleus is active prior to completion of synaptogenesis, we suggest that the facial motoneurons are regulated in a novel or distinct manner during this time period. PMID- 8725307 TI - Immunohistochemical investigation of gamma-aminobutyric acid ontogeny and transient expression in the central nervous system of Xenopus laevis tadpoles. AB - The ontogeny of the gamma-aminobutyric acid (GABA)-positive neurons in the brain of Xenopus laevis tadpoles was investigated by means of immunohistochemistry, using specific antibodies both against GABA and its biosynthetic enzyme, glutamate decarboxylase (GAD). The results obtained with the two antisera were comparable. The GABA system differentiates very early during development. At stages 35/36, numerous GABA-positive neurons were seen throughout the prosencephalon and formed two main bilateral clusters within the lateral walls of the forebrain that ran caudally toward the hindbrain. Other GABA-immunolabeled cell bodies, together with a conspicuous network of GABAergic fibers, were seen in the posterior hypothalamus. In the spinal cord, the lateral marginal zone was GABA-positive, as were Rohon-Beard neurons, interneurons, and Kolmer-Agdhur cells. A very rich GABA innervation was observed in the pars intermedia of the pituitary. At stage 50, plentiful immunopositive neurons and fibers were found in the telencephalic hemispheres, the diencephalon, and the mesencephalon (optic tectum and tegmentum). By stage 54, the number of GABA-immunoreactive neurons in the posterior hypothalamus had decreased, so that, at stage 58, there were very few GABA-labeled cell bodies in the dorsolateral walls of the infundibulum, despite a strong GABAergic innervation within the median eminence and the pars intermedia. From stage 58 to stage 66, the distribution pattern was very similar to that described in the adult X. laevis and in other amphibian species. These results point to transient GABA expression within the hypothalamus, possibly related to either 1) a naturally occurring cell death or 2) a phenotypic switch. PMID- 8725308 TI - Topographic organization in the retinocollicular pathway of the fetal cat demonstrated by retrograde labeling of ganglion cells. AB - The topographic organization of the developing retinocollicular pathway was assessed by making focal deposits of a retrograde tracer (usually rhodamine latex beads) into the superficial layers of the superior colliculus of fetal cats at known gestational ages. Subsequently, the distributions of labeled cells in the contralateral and ipsilateral retinas were examined. At all stages of development, a high density of labeled cells was found in a delimited area (core region) of both retinas. The locations of the retinal regions containing the high density of labeled cells varied with the locus of the tracer deposit in the superior colliculus in a manner consistent with the topographic organization of the mature cat's retinocollicular pathway. Additionally, some labeled ganglion cells, considered to be ectopic, were found to be scattered throughout the contralateral and ipsilateral fetal retinas. Such ectopic cells were few in number throughout prenatal development. For every 100 cells projecting to the appropriate region of the colliculus, we estimate that less than one ganglion cell makes a gross projection error. The incidence of ectopic cells did not differ between the contralateral and ipsilateral retina, even though the overall density of crossed labeled cells was always greater than that of uncrossed labeled cells. In the youngest fetal animals, tracer deposits into the caudal portion of the superior colliculus resulted in a core region of labeled cells in the contralateral nasal retina as well as in the nasal ipsilateral retina. Such uncrossed nasal cells, not seen in more mature animals, appear to innervate the appropriate topographic location of the superior colliculus, but on the wrong side of the brain. Most likely, these uncrossed nasal ganglion cells contribute to the widespread distribution of the ipsilateral retinocollicular pathway observed in fetal cats after intraocular injections of anterograde tracers (Williams and Chalupa, 1982). Collectively, our findings demonstrate that the developing retinocollicular pathway of the fetal cat is characterized by a remarkable degree of topographic precision. PMID- 8725309 TI - Analysis of gene action in the meander tail mutant mouse: examination of cerebellar phenotype and mitotic activity of granule cell neuroblasts. AB - The meander tail (mea) gene results in a stereotypic pattern of cerebellar abnormalities, most notably the virtual depletion of granule cells in the anterior lobe of the cerebellum. The causal basis of this mutation is unknown. In this paper we have taken a three-part approach to the analysis of mea gene action. First, we quantitatively determined the effect of the mea gene on granule cell and Purkinje cell number. We found, in addition to the marked depletion of anterior lobe granule cells ( > 90%), there were also significantly fewer granule cells in the posterior lobe (20-30%) without a concomitant loss of Purkinje cells. Second, we explored the relationship between granule cell depletion caused by the mea gene and by the mitotic poison, 5-fluoro-2'-deoxyuridine (FdU). Prenatal and postnatal ICR mice were treated with FdU to ascertain the regimen that best produces a meander tail-like cerebellar phenotype. The similarity of the effects of the mea gene and injections of FdU at E17 and PO suggests the hypothesis that the mea gene acts to disrupt the cell cycle of cerebellar granule cell precursors. Thus, the third part of this study was to test this hypothesis by using injections of either BrdU (5-bromo-2'-deoxyuridine) or 3H-thymidine into homozygous and heterozygous meander tail littermates at E17 or PO. After processing the tissue for BrdU immunocytochemistry or 3H-thymidine autoradiography, counts were made of the number of labeled and unlabeled external granule layer (EGL) cells to determine the percentage that had incorporated the mitotic label (labeling index). No difference in the labeling index was found between homozygous meander tail mice and normal, heterozygous littermate controls. Therefore, the mitotic activity of the EGL neuroblasts is not disrupted by the mea gene. Furthermore, while a mitotic poison can produce a phenotype similar to the action of the mea gene, mea is phenomenologically different from FdU treatment. PMID- 8725310 TI - Syncope in dilated cardiomyopathy is a predictor of sudden cardiac death. AB - Fifty percent of patients with dilated cardiomyopathy die within 5 years of diagnosis. Syncope is known to be a predictor of poor outcome in patients with advanced heart failure. To assess the risk of patients with dilated cardiomyopathy with a history of syncope during standard medical treatment we compared this group to similar patients without syncope. Twenty-three patients with angiographically proven dilated cardiomyopathy and syncope were followed prospectively and compared to 201 patients without history of syncope. All patients showed a left-ventricular ejection fraction of less than 45%. Both groups did not differ in left-ventricular ejection fraction at baseline (30 +/- 7% in the syncope group, 30 +/- 8% in the no syncope group). Mean follow-up was 2.6 years in the syncope group and 2.4 years in the no syncope group. At baseline, syncope patients used more often amiodarone (p < 0.04), while there was no statistically significant difference between the two groups regarding the intake of digitalis, diuretics and angiotensin-converting enzyme inhibitors. Twenty-six percent of patients in the syncope group and 20% in the no syncope group died during follow-up (non significant). The striking difference, however, was the type of death: 5 out of 6 patients in the syncope group died suddenly compared to 13 of 41 patients in the no syncope group (p < 0.025). Patients with dilated cardiomyopathy and a history of syncope are at high risk of sudden death. PMID- 8725311 TI - Factors predicting maintenance of sinus rhythm after direct current cardioversion of atrial fibrillation and flutter: a reanalysis with recently acquired data. AB - A prospective study was conducted to evaluate how many patients maintain normal sinus rhythm after direct current (DC) cardioversion of atrial arrhythmias and to assess factors predictive of long-term success. The study group consisted of 61 patients (45 men) aged 18-88 years (mean age 66 +/- 11 years) who underwent cardioversion at our department from October 1990 to June 1992. Prior to cardioversion, the patients' medical history, medications, heart size on chest X ray, and echocardiographic findings were reviewed. Overall, 41 (67.2%) patients were in atrial fibrillation, while 20 (32.8%) had atrial flutter. Only 15% of the patients had valvular heart disease. Sinus rhythm was restored by DC cardioversion in 47 (77%) patients, none of whom experienced an embolic event prior to discharge. Patients with atrial flutter had a higher conversion rate (95%) than those in atrial fibrillation (68.3%; p = 0.024), and also patients with an arrhythmia for less than 1 week (94.4%) compared to those with a longer or unknown duration (69.8%; p = 0.047). The primary success rate was not influenced by heart size on chest X ray or echocardiographic variables. The study protocol aimed at following up the patients for 1 year after cardioversion. Of the 47 patients who converted to sinus rhythm data are available on 44 for a mean follow-up of 11 +/- 3 months (range 1-14 months), at which time 25 (57%) still remained in sinus rhythm. Heart size on the chest X ray was significantly increased in the group that did not maintain sinus rhythm (p = 0.03) and their left atrial size on echocardiography was slightly increased (p = 0.10). Patients who originally had atrial flutter were more likely to remain in sinus rhythm than those who had been in atrial fibrillation (p = 0.12), as did patients with an arrhythmia for less than 1 week prior to cardioversion in comparison to those with a longer or unknown duration (p = 0.11). Thus, in contrast to previous reports, according to these recent data on a patient population with a low prevalence of valvular heart disease, DC cardioversion can be attempted in most patients with atrial tachyarrhythmias. Clinical factors, heart size on chest X ray and echocardiographic findings should, however, be considered before deciding to perform DC cardioversion. PMID- 8725312 TI - Increased serum levels of circulating intercellular adhesion molecule-1 in patients with myocarditis. AB - Levels of circulating intercellular adhesion molecule-1 (cICAM-1) were measured by ELISA in the sera of 21 patients with unexplained cardiac dysfunction. By evaluating the biopsy or autopsy specimens, they were classified into two groups: a myocarditis group (n = 8) and a nonmyocarditis group (n = 13). Levels of cICAM 1 were significantly higher in the myocarditis group (p < 0.01). Increased levels of cICAM-1 (> 2 SD above the control mean) were detected in 6 of 8 patients in the myocarditis group. Therefore, the detection of increased levels of cICAM-1 could be a useful marker for myocarditis. PMID- 8725313 TI - Changes in plasma cholesterol levels after hospitalization for acute coronary events. AB - To retrospectively assess the changes in total cholesterol levels after a hospital admission for an acute coronary event, 287 patients were identified who had one isolated event: 130 patients with acute myocardial infarction, 122 patients after coronary artery bypass graft surgery, 35 patients after percutaneous transluminal coronary angioplasty. To be included, patients had to have a total cholesterol measurement within the 3 months prior to hospitalization and periodically after the acute coronary event. Total cholesterol measurements were recorded during four time periods: 0-3 months before hospital admission (baseline); and 0-3, 3-6, and 6-9 months after the hospitalization. Mean total cholesterol value was used if a patient had multiple measurements during a time period. RESULTS: There was no significant difference in the baseline total cholesterol levels among the three groups (acute myocardial infarction, coronary artery bypass graft surgery and percutaneous transluminal coronary angioplasty). In all three groups, there was a decrease in total cholesterol level after hospital admission compared to baseline (p < 0.0001). This decrease was significantly greater in coronary artery bypass graft surgery patients compared to the other two groups. Total cholesterol levels returned to baseline levels by 3 months after the hospitalization in all three groups. Total cholesterol decreased significantly from baseline in patients after an acute myocardial infarction; this decrease was significantly greater if they received thrombolytic therapy (p < 0.05). Total cholesterol returned to baseline 3 months after hospitalization in both groups. CONCLUSIONS: During the 3 months following hospital admission for an acute coronary event, total cholesterol levels are not representative of the patient's baseline values. Management of hypercholesterolemia in this setting requires the use of baseline (preadmission) total cholesterol values. PMID- 8725314 TI - Relationship between autonomic nervous system test results, ejection fraction and inducibility of sustained ventricular arrhythmias by means of electrophysiological studies. AB - Autonomic nervous system dysfunction and low left ventricular ejection fraction have been associated with an increased incidence of cardiac arrhythmias and sudden death. This research was conducted to determine whether Valsalva ratio (longest to shortest R-R interval during Valsalva manoeuvre), R-R variability (standard deviation of R-R intervals in a 60-second electrocardiographic recording strip), and corrected QT interval permit the identification of those patients with an inducible sustained ventricular arrhythmia when submitted to electrophysiological studies. We also evaluated the ejection fraction as a predictive index of inducibility. We studied 42 patients (21 males), 38 +/- 19 years old, who were submitted to an electrophysiological study in order to evaluate supraventricular (control group: n = 16) or ventricular arrhythmias (n = 26). The patients studied for the evaluation of ventricular arrhythmias presented structural and functional cardiac damage. Their ejection fraction (0.41 +/- 0.17) was significantly reduced (p = 0.01) when compared to that of the control group (0.61 +/- 0.08). The patients with cardiac damage had abnormal autonomic nervous system test results. None of the measurements was significantly related to the inducibility of sustained ventricular tachyarrhythmias in the group of patients who had structural and functional involvement. The finding of abnormal cardiac autonomic nervous system indices and low ejection fraction can be related to structural heart disease, but neither the abnormal cardiac autonomic nervous system indices nor the low ejection fraction allows us to identify the patients who will develop inducible sustained ventricular tachyarrhythmias when submitted to electrophysiological study. PMID- 8725315 TI - Restoration of atrial function after atrial fibrillation of different etiological origins. AB - In order to evaluate atrial function after the restoration of normal sinus rhythm in patients with atrial fibrillation, an echocardiographic study was performed in 60 patients hospitalized for atrial fibrillation that was subsequently converted to sinus rhythm. The study population consisted of 60 patients, 41 males and 19 females, with a mean age of 67 +/- 8 years. In etiological terms, 20 patients had atrial fibrillation with no associated pathology, 20 had an underlying ischemic cardiopathy and 20 had hypertension. The duration of atrial fibrillation varied from 24 h to 6 months. The restoration of sinus rhythm was spontaneous in 20 patients and drug induced in 40. Patients were divided into groups on the basis of the duration of the atrial fibrillation, the etiology of the atrial fibrillation and the size of the atrium. Transmitral Doppler inflow patterns at rest were assessed within 24 h after conversion to sinus rhythm, and 1 week, 1 month and 3 months later. The statistical significance of serial changes in peak A velocity and A wave integral was determined by analysis of variance. Using the peak A wave for atrial filling and the velocity-time integral as the index of atrial mechanical function, the following results were obtained: restoration of atrial mechanical function occurred earlier in hypertensive patients and in patients with atrial fibrillation only than in patients with associated ischemic cardiopathy. Restoration also occurred earlier in the group of patients that had atrial fibrillation for < 24 h and in patients whose left atrium was of normal size. Since the mechanical activity of the left atrium, evaluated as the peak A wave, does not necessarily return immediately after the restoration of sinus rhythm, a follow-up assessment of the contraction of the left atrium is appropriate. PMID- 8725316 TI - Hemodynamic comparison of Medtronic intact bioprostheses and bileaflet mechanical prostheses in aortic position. AB - Hemodynamic properties of both bioprostheses and mechanical valves have improved over the years, however, direct comparison between second-generation porcine and bileaflet valves is rare. Therefore, by the mean of Doppler echocardiography, we determined hemodynamic performances in 45 patients with Medtronic Intact bioprostheses in the aortic position and in 67 patients with bileaflet mechanical valves. Differences in pressure gradients between the two groups were essentially attributable to differences in annulus size. Indeed when the transvalvular gradient was stratified according to annulus size, mean values were no longer significantly different between Medtronic Intact porcine valves and bileaflet mechanical valves for each annulus size. In conclusion, Medtronic Intact bioprostheses in the aortic position exhibit similar hemodynamic characteristics compared to bileaflet mechanical valves. PMID- 8725317 TI - A mechanism of ischemic preconditioning during percutaneous transluminal coronary angioplasty. AB - Manifestation of ischemic preconditioning and its mechanisms during percutaneous transluminal coronary angioplasty (PTCA) was evaluated. Twenty-two patients with angina pectoris, who had one-vessel coronary artery disease of the proximal left anterior descending artery but without visual collateral circulation, underwent elective PTCA performed by balloon inflations of 90 s, repeated three times or more. Changes in standard 12-lead electrocardiogram, hemodynamics and oxygen saturation of the great cardiac vein by a fiber-optic catheter were analyzed. Anginal chest pain occurred in 21 patients (95%) during the first balloon inflation, and in only 9 patients (41%) during the third inflation. In comparison with the first inflation, the third produced less shifts in ST junction (p < 0.01) and peak T (p < 0.01), which were measured and averaged by 4 chest leads from V2 to V5. The heart rate-blood pressure product during the third inflation was equivalent to that during the first. The great cardiac vein oxygen saturation decreased equally during the first and third inflations. However, the ratio of the saturation at reactive hyperemia after balloon deflation to baseline was higher (p < 0.01) in the third than in the first inflation. The adenosine content of the great cardiac vein measured in 11 patients just prior to balloon deflation was also higher (p < 0.05) in the third inflation than the value in the first. Repeated coronary artery occlusion during PTCA could cause ischemic preconditioning, which may be derived from mechanisms common to accelerated reactive hyperemia, for example an increase in intrinsic adenosine levels. PMID- 8725318 TI - Relationship of pulmonary venous flow pattern to mean left atrial pressure and phasic pressure change. AB - To assess the correlation of pulmonary venous flow pattern with the magnitude of the dynamic change in left atrial pressure (i.e. x and y descents) and to elucidate the way how pulmonary venous flow pattern reflects left atrial pressure, we studied 35 patients with normal sinus rhythm undergoing transesophageal echocardiography and cardiac catheterization. The ratio of peak systolic/diastolic flow correlated well with the ratio of x/y descent (r = 0.88) and the ratio of systolic/diastolic velocity-time integral (r = 0.89). The correlation between the ratio of systolic/diastolic flow or velocity-time integral and mean atrial pressure was less strong (r = -0.49 and -0.70, respectively). We conclude that the ratio of peak systolic/diastolic flow velocity, or the ratio of systolic/diastolic velocity-time integral of pulmonary venous flow reflects the phasic change in left atrial pressure more appropriately than the mean left atrial pressure. Due to the good correlation between the phasic change in left atrial pressure and pulmonary venous flow pattern, the flow pattern of pulmonary veins could be used to identify diseases with specific patterns of left atrial pressure. PMID- 8725319 TI - Treatment of patients with acute myocardial infarction in relation to gender. AB - The question whether women and men with acute myocardial infarction (AMI) are treated differently is currently debated. In this analysis we compared pharmacological treatments and revascularization procedures during hospitalization and during 1 year of follow-up in 300 women and 621 men who suffered an AMI in 1986 or 1987 at our hospital. During hospitalization, the mean dose of morphine (+/- SD) during the first 3 days was higher in men compared to women (14.5 +/- 15.7 vs. 9.8 +/- 10.3 mg, p < 0.001), more men than women were given morphine after the first 24 h (65.4 vs. 49.0%, p < 0.01), and more men were prescribed anticoagulants at discharge (18 vs. 12%, p < 0.05). After 1 year more women than men were on diuretics (61.3 vs. 42.8%, p < 0.001) and a similar observation was made at discharge. This study was performed before thrombolytic therapy was routinely used. The frequency of revascularization procedures did not differ between men and women during hospitalization or during the year of follow up. In conclusion, no major treatment differences, which could affect the prognosis, were found between women and men hospitalized due to AMI in this study in the prethrombolytic era. PMID- 8725320 TI - Persistent thallium-201 defect: can clinical, electrocardiographic and exercise hemodynamic variables predict defect normalization with reinjection? AB - This study was designed to assess the contribution of clinical, electrocardiographic and exercise hemodynamic variables to the prediction of normalization on resting reinjection scintigraphy of persistent thallium-201 (201Tl) myocardial perfusion defects seen with exercise and 2- to 4-hour delayed (redistribution) imaging. To evaluate this contribution, we studied 159 consecutive patients with persistent 201Tl myocardial perfusion defects on routine exercise and 2- to 4-hour-delayed scintigrams at the University of Rochester Medical Center who were classified as having moderate or greater ischemic normalization (group 1, n = 76) or minimal to no ischemic normalization (group 2, n = 83) by reinjection scintigraphy. Multiple logistic regression analysis with backward elimination was used to model the effects of clinical, electrocardiographic and exercise hemodynamic data on the odds ratio of a normalized defect. No difference was observed in the two groups with regard to gender, angina on exertion, rate-pressure product, exercise duration, resting or exertional ischemic ST changes on electrocardiogram, presence of Q waves or left ventricular hypertrophy on baseline electrocardiogram, or total number of stress thallium defects (2.8 +/- 1.5 segments). No single variable or combination of variables discriminated between groups 1 and 2 by logistic regression analysis. We conclude that defect normalization seen on resting 201Tl myocardial perfusion scintigraphy is prevalent in patients with persistent defects on routine exercise and delayed myocardial perfusion scintigraphy, and was not predictable from available clinical, electrocardiographic and exercise hemodynamic variables. PMID- 8725321 TI - Predictive value of sequential testing in screening for silent myocardial ischemia in asymptomatic middle-aged men (the ECCIS Project). AB - The accuracy of sequential testing in the noninvasive diagnosis of coronary artery disease has been established in the symptomatic clinical populations, while little is known about its value when applied to low prevalence groups, such as totally asymptomatic men. To evaluate the accuracy of noninvasive sequential testing in the diagnosis of silent myocardial ischemia, data were collected from exercise electrocardiogram, 201Tl perfusion scintigraphy and radionuclide angiography for 62 totally asymptomatic middle-aged men who underwent coronary arteriography because they were positive for two or more markers of myocardial ischemia as determined by a diagnostic screening of a nonbiased population consisting of 4,842 presumably healthy men aged 40-59 years (the ECCIS Project). The predictive value of serial testing procedures for significant coronary artery obstruction was 35%. Predictive values of an abnormal electrocardiogram associated with either an abnormal 201Tl scintigram, an abnormal isotopic ventriculography, or both were 33, 38 and 31%, respectively. In asymptomatic middle-aged men, there is at least a 50% likelihood that an abnormal radionuclide test is a false-positive result, and the positive predictive value is not enhanced by the concordance of an abnormal 201Tl scintigraphy with an abnormal isotopic ventriculography. Thus, the application of noninvasive sequential testing in screening for asymptomatic coronary artery disease is limited by its low predictive value in accordance with the Bayesian probability theory. PMID- 8725322 TI - Dobutamine stress echocardiography for detecting coronary artery disease. AB - To assess the value of dobutamine stress echocardiography (DSE) to detect coronary artery disease (CAD) and to compare the diagnostic accuracy between DSE and treadmill exercise test (TXT), 104 patients (mean age 58 +/- 12 years) presenting for coronary angiography were prospectively studied. TXT was performed according to the Bruce protocol. Dobutamine (5-40 micrograms/kg/min) was infused in 3-min stages. Digital echocardiograms were recorded on-line at baseline, during low- and peak-dose dobutamine infusion, and at recovery. An echocardiogram positive for CAD was defined as the one showing a new wall motion abnormality (WMA) induced by dobutamine. There were no major complications during the study. Significant CAD (> or = 50% diameter stenosis) was present in 17 of 30 patients who had normal echocardiograms at baseline. The sensitivity for detecting CAD was 76% by TXT and 94% by DSE, and the specificity was only 38% by TXT and 92% by DSE, respectively. Seventy-four patients had localized rest WMAs. Twenty-four had no significant CAD or lesions only confined to regions with abnormal rest wall motion, and 50 had disease remote from these regions. The sensitivity for detection of remote disease was 60% by TXT and 76% by DSE, and the specificity was 75% by TXT and 96% by DSE, respectively. In conclusion, DSE is a safe and accurate diagnostic tool for identifying CAD and for predicting the extent of disease in those who have localized rest WMAs. PMID- 8725323 TI - Prognostic value of echocardiography in 190 patients with chronic congestive heart failure. A comparison with New York Heart Association functional classes and radionuclide ventriculography. AB - Survival in 190 consecutive patients with congestive heart failure, discharged from a general hospital, was studied. Sixteen patients were in New York Heart Association (NYHA) class I, 87 in II, 83 in III and 4 in IV. Median left ventricular ejection fraction (LVEF) from radionuclide ventriculography was 0.30 (range 0.06-0.74). Two-year survival was 68%. Wall motion index was the only echocardiographic variable with significant, independent, prognostic information on survival. The 2-year survival in NYHA classes I and II was 90.7% for wall motion index > or = 1.3, and 78.6% when < 1.3. In classes III and IV survival was 68.9% for wall motion index > or = 1.3 and 39.9% when < 1.3. Addition of LVEF gave further information about survival. This study demonstrates that echocardiography is of great value in determining prognosis in congestive heart failure patients, and that wall motion index contains the majority of the information. Wall motion index is closely correlated to LVEF, however prognostication is improved when information about LVEF is added. PMID- 8725324 TI - Managing acute myocardial infarction in a renal transplant recipient. AB - We describe the management of a patient, with a 13-year-old cadaveric renal transplant, who presented with acute myocardial infarction. Successful primary angioplasty was performed to the left anterior descending artery. It was complicated by transient renal failure and pseudoaneurysm of the femoral artery which was managed conservatively. PMID- 8725325 TI - Treatment of anomalous origin of the left coronary artery from the pulmonary artery in adulthood. AB - A 47-year-old female patient underwent surgical correction of a recently diagnosed anomalous left coronary artery. The artery originated from the pulmonary artery, and the patient had suffered from pulmonary hypertension and congestive heart failure. The follow-up after 3 and 6 months showed only slight improvement in the patient's condition. PMID- 8725326 TI - Congenital coronary artery fistulae arising from bilateral coronary arteries and emptying into both pulmonary artery and left ventricle: a rare presentation. AB - Congenital coronary artery fistulae rarely empty into both the pulmonary artery and the left ventricle. To our knowledge, there have been only two such cases reported in the literature. This paper presents a third case. The pattern of drainage into the left ventricle in the present case was, however, quite different from that observed in the previous reports. PMID- 8725327 TI - False-positive cardiac troponin T levels in chronic hemodialysis patients. PMID- 8725328 TI - Nomenclature problems in modern cardiology. PMID- 8725329 TI - [Three-dimensional images of the cystic duct obtained with helical DIC CT]. AB - Before laparoscopic cholecystectomy, it is important to clarify the anatomy of the cystic duct. This study assessed three-dimensional CT images (3D images) of the cystic duct obtained non-invasively using helical DIC CT and these images were compared with those obtained with ERCP and DIC. The three-dimensional technique using Helical DIC-CT was applied in 168 patients for laparoscopic cholecystectomy. The cystic duct detected by 3D imaging was evaluated for patency, length and bifurcation. Three-dimensional images showed the cystic duct in 157 of 168 cases (93.5%) and in 81 of 89 cases (91%) in which the cystic duct was not clearly visualized on DIC. Among the 23 cases in which were both 3D images and ERCP undergone, 3D images were equal to those of ERCP in detection of the cystic duct in 20 cases, superior to ERCP in two cases, and inferior to ERCP in one. The technique of 3D images proved useful in demonstrating the patency, length and variations in bifurcation of the cystic duct for surgeons performing laparoscopic cholecystectomy and might be substitute ERCP in preoperative assessment. PMID- 8725330 TI - [CT diagnosis of acute occlusion of the superior mesenteric artery]. AB - The purpose of this study was to clarify the usefulness of CT in the early diagnosis of acute SMA occlusion. The CT scans of seven patients with surgically proved acute SMA occlusion were retrospectively reviewed. The SMV/SMA ratio, which was defined as the external diameter of the SMV divided by the external diameter of the SMA, was measured on CT scans in seven patients and in 300 control subjects. The sites of SMA occlusion were seen as high density structures on preconstrast CT scans in all seven patients and as filling defects on postcontrast CT scans in five of them. According to pathologic and angiographic studies, the high density structures represented intravascular fresh blood clots. The SMV/SMA ratios were < 1 in five of seven patients, whereas they were > 1 in 288 of 300 control subjects. Other CT findings, such as dilated fluid-filled bowel, decreased enhancement of small bowel wall and ascites, seemed to be dependent on the extent of bowel necrosis. In conclusion, focal high density in the SMA on preconstrast CT scan is a specific sign of acute SMA occlusion, and the SMV/SMA ratio might reflect the ischemic grade of mesenteric circulation. PMID- 8725331 TI - [Analytical evaluation of compression and displacement of the esophagus and trachea due to right aortic arch with MR imaging]. AB - We studied 12 cases of right aortic arch (RAA) with MRI, and the anatomical relationship between right aortic arch and the esophagus and trachea were analysed. Three of 12 cases showed RAA with mirror-image branching. Nine cases were RAA with aberrant left subclavian artery. The proximal portion of the aberrant left subclavian artery in the retrotracheoesophageal space was expanded just like a pouch. The pouch was the 8th segment dorsal aortic root and the so called aortic diverticulum. This was one of the causes of symptoms such as wheezing and dysphagia. We divided the configuration of aortic diverticulum into two types, bulging type and saccular type, on the basis of coronal images. Two cases were bulging type, seven saccular type. The axial images in two cases showed compressed esophagus, and in three cases showed displacement of the trachea, narrowing of the space like a triangle surrounded by the aortic arch, aortic diverticulum and aberrant left subclavian artery. We drew lines and measured the angle of the aortic arch with the aberrant left subclavian artery on the axial images of these cases. We found that cases with an angle of less than 60 degrees showed a high correlation with the causes of compression of the esophagus or displacement of the trachea. It was necessary to follow such patients with MRI. PMID- 8725332 TI - [Multidirectional pseudodynamic studies of the bilateral temporomandibular joint diseases with 0.2-Tesla MR imaging unit]. PMID- 8725333 TI - [Classification of US findings of ovarian masses in children]. AB - The ultrasonographic findings of 25 lesions in 23 patients with surgically proven ovarian masses were reviewed. There were 10 cystic teratomas, two simple cysts, two follicular cysts, two mucinous cystadenomas, two NHL, one corpus luteum cyst, one hydrosalpinx, one serous cystadenoma, one yolk sac carcinoma, one dysgerminoma, one embryonal carcinoma, and one mixed form (yolk sac carcinoma, choriocarcinoma). All patients were less than 15 years old. We classified all cases into four patterns: cystic, cystic with mural nodule, mixed, and solid. Eight lesions of the cystic pattern included two simple cysts, two follicular, cysts and one corpus luteum cyst. The other lesions were benign, too. Nine lesions with the cystic with mural nodule pattern consisted of eight cystic teratomas and one mucinous cystadenoma. All lesions were benign. The mixed pattern was seen in four lesions, half of which were malignant, i. e., one embryonal carcinoma and one yolk sac carcinoma. Four lesions with the solid pattern were all malignant masses: one dysgerminoma, two NHL and one mixed form. In this classification, the cystic and cystic with mural nodule patterns are benign, while mixed and solid patterns are highly suggestive of malignancy. PMID- 8725334 TI - [CT findings of the small polypoid lesions of the gallbladder (2cm or less PLG): differentiation between benign and malignant disease on unenhanced CT]. AB - Recently, the detection of polypoid lesions of the gallbladder (PLG), by using ultrasonography has been increased. Nevertheless, it is difficult to distinguish between benign PLG and malignant PLG. The purpose of this paper is to evaluate the efficacy of CT examination for differential diagnosis between benign and malignant PLG. Twenty-nine patients with PLG diagnosed by preoperative CT were retrospectively reviewed. Out of these patients, 20 patients had benign PLG and nine patients had malignant PLG. As a result, all PLG were well enhanced by contrast media. On unenhanced CT, malignant PLG appeared to be high density except one case. On the other hand, benign PLG were isodense except one case. Relatively larger amount of epithelial component of the malignant PLG contributes to high density on unenhanced CT. In contrast, benign PLG had larger amount of connective tissue, showing about the same density as bile on unenhanced CT. In conclusion, unenhanced CT examination is effective to differentiate benign PLG from malignant PLG. PMID- 8725335 TI - [Usefulness of multi-plane dynamic subtraction CT (MPDS-CT) for intracranial high density lesions]. AB - We present a new CT technique using the high speed CT scanner in detection and evaluation of temporal and spatial contrast enhancement of intracranial high density lesions. A Multi-Plane Dynamic Subtraction CT (MPDS-CT) was performed in 21 patients with intracranial high density lesions. These lesions consisted of 10 brain tumors, 7 intracerebral hemorrhages and 4 vascular malformations (2 untreated, 2 post embolization). Baseline study was first performed, 5 sequential planes of covering total high density lesions were selected. After obtaining the 5 sequential CT images as mask images, three series of multi-plane dynamic CT were performed for the same 5 planes with a intravenous bolus injection of contrast medium. MPDS-CT images were reconstructed by subtracting dynamic CT images from the mask ones. MPDS-CT were compared with conventional contrast enhanced CT. MPDS-CT images showed the definite contrast enhancement of high density brain tumors and vascular malformations which were not clearly identified on conventional contrast-enhanced CT images because of calcified or hemorrhagic lesions and embolic materials, enabling us to except enhanced abnormalities with non-enhanced areas such as unusual intracerebral hemorrhages. MPDS-CT will provide us further accurate and objective information and will be greatly helpful for interpreting pathophysiologic condition. PMID- 8725336 TI - [Results of radiotherapy combined with CDDP (BAI) plus VP-16 in the treatment of inoperable non-small cell lung cancer]. AB - Twenty-six patients with unresectable non-small cell lung cancer were treated by chemotherapy alone or chemotherapy followed by radiotherapy from June 1988 to May 1990. CDDP (80 mg/m2) was given by bronchial arterial infusion (BAI) on day 1 in each course, VP-16 (80 mg/body) was given by drip intravenous infusion on days 1 and 2, and VP-16 (150 mg/body) was given orally on days 3 and 5. The patients were irradiated after this regimen with a single dose of 1.8-2.0 Gy, five times each week, to a volume which encompassed only the primary tumor. A total dose of 60 Gy or more was delivered. There were 10 squamous cell carcinomas, nine adenocarcinomas, five large cell carcinomas, one adenosquamous cell carcinoma and one poorly differentiated carcinoma. Median survival time (MST) was 354 days in stage III B patients and 280 days in stage IV patients. MST was 155 days in adenocarcinoma and 310 days in squamous cell carcinoma. MST was 372 days in PS 0 to 1 patients and 140 days in PS 2 patients. With chemo-radiotherapy in 17 cases resulted in no complete remissions and 10 partial remissions. The overall response rate was 58.8%. It is concluded that CDDP via BAI combined with VP-16 administration and sequential irradiation is useful to acquire a high response rate in non-small cell lung cancer. However, further studies on this modality are required. PMID- 8725337 TI - [Post-therapeutic changes in body composition of patients with Graves' disease]. AB - Changes in patients' body composition following therapy for Graves' disease were assessed by dual energy X-ray absorptiometry (DXA). The subjects were 21 patients (6 males, 15 females) who were treated with 131I. Their ages ranged from 24 to 75 years (median 51.2 years). Changes in body composition were analyzed by evaluating the fat mass (FM) and lean mass (LM). Both FM and LM were significantly increased after therapy. The increase in LM was greater in the males, whereas in females, no significant difference was noted between the changes in FM and LM. The distributions of FM and LM were examined in the arms, legs and trunk after therapy, and FM was significantly increased in the trunk, while LM was increased in the legs. DXA is useful for the assessment of body composition as well as bone mineral content in follow-up study of Graves' disease. PMID- 8725338 TI - [Fast two-compartment model analysis with 99mTc-GSA liver scintigraphy]. AB - The use of numerical integration method (N. INT) was evaluated in determining parameters by two-compartment model analysis from liver scintigraphy. Among the 15 subjects, 14 had liver cirrhosis or chronic hepatitis, and one was normal. When using N.INT, the sum of two exponential functions on heart regression curve following intravenous injection of 99mTc-galactosyl human serum albumin (99mTc GSA) was promptly calculated. The parameters obtained from N.INT, including transfer rate of 99mTc-GSA from extrahepatic blood to liver (k1), dissociation rate from liver to extrahepatic blood (k2), and excretion rate from liver to gallblader (k3), were significantly correlated with those obtained by the nonlinear least square method (NLS). k1/k2 was related with the maximum removal rate of 99mTc-GSA obtained from nonlinear five compartment model analysis (r2 = 0.705, n = 15) and also with the severity score of liver disease as classified by the First Department of Surgery, Mie University Medical School (r2 = 0.686, n = 13). In terms of the time required to obtain these parameters, including the blood retention rate of 99mTc-GSA at 15 min after injection (%ID15), N.INT was faster than the traditional method (NLS). PMID- 8725339 TI - [Measurement of gastric emptying by magnetic resonance imaging]. AB - The purpose of the study was to establish a new method of measuring gastric emptying using MR Imaging in human. Gastric emptying was measured in 6 healthy male volunteers aged from 28 to 43 years, using MR imaging and RI. The measurements were performed after the oral administration of liquid meal containing glucose, protein and fat. The MR imaging was performed with 0.5T superconducting magnet machine, and consecutive 12 transaxial T1 weighted spin echo images (TR/TE = 300/17) of the upper abdomen were recorded every 10 minutes for more than 1 hour. Gastric emptying curves and their T1/2 values obtained by MR imaging and RI method were correlated well in 5 of 6 cases. We concluded that a non invasive and radiation free method using MR imaging was proved to be a useful tool for measuring gastric emptying. PMID- 8725340 TI - [Virtual CT endoscopy "Cruising Eye View": development and clinical applications]. AB - We have developed a virtual CT endoscopy, designated as "Cruising Eye View (CEV)", which have generated from spiral volumetric CT data and reconstracted using a new three-dimensional image reconstruction technique. Using this new method, we have reconstructed the inner image of patients with aortic dissection (2 cases) and arteriosclerosis obliterance (3 cases). The view point and view direction could be set arbitrarily in the vessel, so that the site, configuration, and/or degree of the intravascular abnormalities were recognized at first glance. Virtual CT endoscopy with CEV method is a promising, non invasive strategy for archiving the inner image of diverse aorto-arterial pathologies. PMID- 8725341 TI - [Autoradiographic study of myocardial fatty acid metabolism in diabetic mouse using 125I-BMIPP]. AB - The existence of diabetic cardiomyopathy has been substantiated in patients with diabetes mellitus. In this study we evaluated two mice with diabetes mellites (DM) and two control mice to determine whether the fatty acid metabolism of the myocardium is abnormal when 125I-BMIPP is used. 125I-BMIPP of 0.74 MBq (20 microCi) was injected, and 30 min later, the mice were slaughtered. Heart, lungs, liver, spleen and kidneys were removed. After freezing the heart, myocardium was sliced into 20 micron sections using a cryostat. Each slice was placed in contact with an imaging plate for autoradiography. A bioimaging analyzer was used to analyze the data. The percent injected dose/g of heart was lower in the DM group than in the control group (11.6 +/- 0.49 vs 18.3 +/- 1.27). Each slice of the DM group also had fewer counts than slices in the control group (p < 0.05 or p < 0.01). These results indicated that fatty acid metabolism in the myocardium was less in the DM group than in the control group. PMID- 8725342 TI - Expression of a phosphorylated isoform of MAP1B is maintained in adult central nervous system areas that retain capacity for structural plasticity. AB - Microtubule-associated protein IB (MAP1B) is the first MAP to be detected in the developing nervous system, and it becomes markedly down-regulated postnatally. Its expression, particularly that of its phosphorylated isoform, is associated with axonal growth. To determine whether adult central nervous system (CNS) areas that retain immunoreactivity for MAP1B are associated with morphological plasticity, we compared the distribution of a phosphorylated MAP1B isoform (MAP1B P) to the distribution of total MAP1B protein and MAP1B-mRNA. Although they were present only at very low levels, both protein and message were found ubiquitously in almost all adult CNS neurons. The intensity of staining, however, varied markedly among different regions, with only a few nuclei retaining relatively high levels. MAP1B-P was restricted to axons, whereas total MAP1B was present in cell bodies and processes. Relatively to total MAP1B protein and its mRNA, MAP1B P levels decreased more dramatically with maturation, and they were detectable in only a few specific areas that underwent structural modifications. These included primary afferents and motor neurons, olfactory tubercles, habenular and raphe projections to interpeduncular nuclei, septum, and the hypothalamus. The distribution pattern of MAP1B-P was compared to that of the embryonic N-CAM rich in polysialic acid (PSA-NCAM). We found that the PSA-NCAM immunostaining was largely overlapped with that of MAP1B-P in the adult CNS. These results suggest that, like PSA-NCAM, MAP1B may be one of the molecules expressed during brain development that also plays a role in structural remodeling in the adult. PMID- 8725343 TI - Regional and laminar differences in synaptic localization of NMDA receptor subunit NR1 splice variants in rat visual cortex and hippocampus. AB - Changes in N-methyl-D-aspartate (NMDA) receptor expression may represent a molecular substrate for differences in synaptic plasticity between early postnatal and adult brains (Fox and Zahs [1994] Curr. Opinion Neurobiol. 4:112 119). We have, therefore, examined the regional and laminar distribution of NR1, the essential subunit of the NMDA receptor, in two regions in which synaptic plasticity has been most thoroughly studied: primary visual cortex and hippocampus. To study NR1 expression at the light and electron microscopic levels we have used a new antiserum (NR1-C1; Sheng et al. [1994] Nature 368:144-147) directed against a differentially spliced C-terminal exon ("C1"). The most striking result was that the pattern of NR1-C1 labeling in the adult was more restricted than that of previously published NR1-specific antibodies. Specifically, NR1-C1 did not label cells in the CA3, dentate gyrus or subicular regions of the hippocampus or in layer 4 of the visual cortex. Quantitative ultrastructural analysis revealed that these differences were paralleled by differential expression of NR1-C1 at synapses. In sharp contrast to the pattern in the adult, NR1-C1 immunoreactivity was distributed more widely in the developing brain. At postnatal day 11, NR1-C1 splice variants were expressed in all layers of the visual cortex and in all regions of the hippocampus. The transient expression of NR1-C1 splice variants in layer 4 of visual cortex suggests that NR1-C1 may play a role in determining the critical period for binocular plasticity. Continued expression of NR1-C1 in upper and lower layers of the adult cortex and in CA1 of the hippocampus may provide a substrate for plasticity in corticocortical connections and Schaffer collateral synapses beyond the critical period. In addition to abundant postsynaptic staining, NR1-C1 immunoreactivity was found in a large number of axon terminals in the dorsal subiculum, but in very few terminals in visual cortex. This strongly suggests that presynaptic NMDA receptors play a major role in neuronal processing of hippocampal output through the subiculum, but play a relatively minor role in visual processing. PMID- 8725344 TI - Differential expression of annexins I-VI in the rat dorsal root ganglia and spinal cord. AB - The annexins are a family of Ca(2+)-dependent phospholipid-binding proteins. In the present study, the spatial expression patterns of annexins I-VI were evaluated in the rat dorsal root ganglia (DRG) and spinal cord (SC) by using indirect immunofluorescence. Annexin I is expressed in small sensory neurons of the DRG, by most neurons of the SC, and by ependymal cells lining the central canal. Annexin II is expressed by most sensory neurons of the DRG but is primarily expressed in the SC by glial cells. Annexin III is expressed by most sensory neurons, regardless of size, by endothelial cells lining the blood vessels, and by the perineurium. In the SC, annexin III is primarily expressed by astrocytes. In the DRG and the SC, annexin IV is primarily expressed by glial cells and at lower levels by neurons. In the DRG, annexin V is expressed in relatively high concentrations in small sensory neurons in contrast to the SC, where it is expressed mainly by ependymal cells and by small-diameter axons located in the superficial laminae of the dorsal horn areas. Annexin VI is differentially expressed by sensory neurons of the DRG, being more concentrated in small neurons. In the SC, annexin VI has the most striking distribution. It is concentrated subjacent to the plasma membrane of motor neurons and their processes. The differential localization pattern of annexins in cells of the SC and DRG could reflect their individual biological roles in Ca(2+)-signal transduction within the central nervous system. PMID- 8725345 TI - Unilateral LTP triggers bilateral increases in hippocampal neurotrophin and trk receptor mRNA expression in behaving rats: evidence for interhemispheric communication. AB - Induction of long-term potentiation (LTP) in the dentate gyrus of awake rats triggered a rapid (2 hour) elevation in tyrosine kinase receptor (trkB and trkC) gene expression and a delayed (6-24 hour) increase in brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NT-3) gene expression. Depending on the mRNA species, LTP induction led to highly selective unilateral or bilateral increases in gene expression. Specifically, trkB and NT-3 mRNA elevations were restricted to granule cells in the ipsilateral dentate gyrus, whereas bilateral increases in trkC, BDNF, and nerve growth factor (NGF) mRNA levels occurred in granule cells and hippocampal pyramidal cells. Both unilateral and bilateral changes in gene expression were N-methyl-D-aspartate (NMDA) receptor-dependent and LTP-specific. Bilateral electrophysiological recordings demonstrated that LTP was unilaterally induced; this was corroborated by a dramatic unilateral increase in the expression of the immediate early gene zif/268, a marker for LTP, restricted to the ipsilateral granule cells. The results indicate that LTP triggers an interhemispheric communication manifested as selective, bilateral increases in gene expression at multiple sites in the hippocampal network. Furthermore, our findings suggest that physiological plastic changes in the adult brain may involve coordinated, time-dependent regulation of multiple neurotrophin and trk receptor genes. PMID- 8725346 TI - Microcircuitry of forward and feedback connections within rat visual cortex. AB - Visual cortex in mammals is composed of many distinct areas that are linked by reciprocal connections to form a multilevel hierarchy. Ascending information is sent via forward connections from lower to higher areas and is thought to contribute to the emergence of increasingly complex receptive field properties at higher levels. Descending signals are transmitted via feedback connections from higher to lower areas and are believed to provide information about the context in which a stimulus appears, to contribute to modulation of visual responses by attention, and to play a role in memory processes. To determine whether forward and feedback pathways in rat visual cortex constitute distinct intracortical circuits, we have studied the distribution of reciprocal corticocortical inputs to pyramidal cells and gamma-aminobutyric acid (GABA)ergic interneurons. For this purpose, we chose forward and feedback connections between primary visual cortex and the secondary extrastriate lateromedial (LM) area as a model system. Pathways were traced with the axonal marker phaseolus vulgaris-leucoagglutinin. Labeled terminals were identified in the electron microscope, and GABA immunocytochemistry was used to identify the postsynaptic dendritic shafts of GABAergic interneurons. In both pathways, inputs to pyramidal cells were directed preferentially to dendritic spines and not to shafts. In the forward pathway, 90% of labeled inputs were distributed to pyramidal cells and 10% to interneurons. This proportion was similar to that of nearby unlabeled connections in the neuropil, indicating that forward connections are not selective for pyramidal cells or interneurons. In sharp contrast, feedback connections were significantly different from the unlabeled connections and supplied almost exclusively pyramidal cells (98%). Feedback inputs to GABAergic neurons were five times weaker (2%) relative to the forward direction. These structural differences suggest that disynaptic GABAergic inhibition is much stronger in forward than in feedback pathways. Recent physiological experiments have confirmed this prediction (Shao et al. [1995] Soc. Neurosci. Abstr., 21:1274) and we, therefore, conclude that relatively small anatomical differences in the microcircuitry can have important functional consequences. It remains an open question whether generally reciprocal interareal circuits at all levels of the cortical hierarchy are organized in similar fashion. PMID- 8725347 TI - GluR1-immunopositive interneurons in rat neocortex. AB - Recent in vitro studies suggest that inhibitory interneurons in cortex may express the GluR1 glutamate receptor subunit in the absence of GluR2, leading to calcium-permeable alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA) channels. We performed a study of rat somatic sensory cortex to confirm and extend these observations, using quantitative immunocytochemistry for multiple antigens. A morphologically distinct subpopulation of nonpyramidal neurons in neocortex was intensely immunoreactive for GluR1. Electron microscopic analysis of these cells revealed somatic staining for GluR1, mainly in the rough endoplasmic reticulum. Dendritic staining was concentrated at the synaptic active zone and in the adjacent subsynaptic cytoplasm. Double immunostaining revealed that the large majority of intensely GluR1-positive cells contained gamma aminobutyric acid or its synthetic enzyme, glutamic acid decarboxylase, but little or no GluR2. Thus, AMPA receptors on a subpopulation of inhibitory interneurons in cortex are likely to be calcium permeable. This calcium permeability is likely to influence functional properties of these neurons; it may underlie the high levels of calcium-binding proteins they contain; and may render them liable to excitotoxic injury PMID- 8725348 TI - Synapse restructuring associated with the maintenance phase of hippocampal long term potentiation. AB - Synapses in the middle molecular layer of the rat dentate gyrus were analyzed by electron microscopy during the maintenance phase of long-term potentiation (LTP). LTP was induced by high-frequency stimulation of the medial perforant path carried out on each of 4 consecutive days. The dentate gyrus was examined electron microscopically 13 days following the fourth stimulation. At this time point, synaptic responses were still significantly enhanced relative to baseline, although the extent of their potentiation was lower than 1 hour after the last high-frequency stimulation. Stimulated, but not potentiated, rats served as controls. Using the stereological double disector method, estimates of the number of different morphological types of synapses per postsynaptic neuron were obtained. The number of asymmetrical axodendritic synapses increased (by 28%) during LTP maintenance, whereas the number of other synaptic types was not significantly altered. Our previous work demonstrated that the induction of LTP is followed by a selective increase in the number of axospinous perforated synapses with multiple, completely partitioned, transmission zones. Thus, the induction and maintenance phases of LTP are characterized by different structural synaptic alterations. These alterations may be related to each other as indicated by another finding of the present study regarding the existence of perforated synapses that appear to be transitional between axospinous and axodendritic junctions. This suggests a model of structural synaptic plasticity associated with LTP in which some axospinous perforated synapses increase in numbers shortly after the induction of LTP and are then converted into axodendritic ones during LTP maintenance. PMID- 8725349 TI - Projection of the nucleus pretectalis to a retinorecipient tectal layer in the pigeon (Columba livia). AB - The avian optic tectum is composed of at least 15 separate laminae that are distinguishable on the basis of their morphological features and patterns of afferent and efferent connectivity. Layer 5b, a major retinorecipient layer, exhibits dense, dust-like, neuropeptide Y-positive (NPY+) immunoreactive labeling, whereas sparse, larger caliber NPY+ fibers are found in laminae 4 and 7. Anterograde and retrograde labeling techniques, immunohistochemistry, and retinal lesion studies were used to determine the source of this tectal NPY+ labeling. NPY+ was not detectable in cells of the optic tectum or in retinal ganglion cells, and retinal ablation did not diminish the abundance of tectal NPY+ fibers. Neurons of two nuclei previously shown to be sources of tectal input, the nucleus pretectalis (PT) and the intergeniculate leaflet (IGL; Brecha, 1978), were found to be NPY+. Unilateral injection of retrograde tracers into the tectum resulted in bilateral labeling of neurons within PT, and injections of anterograde tracer into PT confirmed that this nucleus projected bilaterally to layer 5b of the optic tectum. Unilateral lesions of PT nearly eliminated NPY+ fibers in the ipsilateral layer 5b and significantly reduced them in the contralateral layer 5b. Bilateral lesions of PT eliminated NPY+ fibers bilaterally in layer 5b. However, these PT lesions had little effect on the NPY+ fibers in layers 4 and 7. Combined retrograde and immunohistochemical studies showed that NPY+ neurons of the IGL project to the optic tectum, and anterograde studies demonstrated that IGL projects to layers 4 and 7. The NPY+ projection to laminae 5b from PT is one of many inputs, which include cholinergic afferents from the nucleus isthmi parvicellularis, terminals from retinal ganglion cells, and dendrites of layer 13 neurons (Karten et al., 1993). The NPY+ input to layer 5b may modulate visual information flow from retinal input to various tectal neurons, including those in layer 13. PMID- 8725350 TI - Cortical lesions induce an increase in cell number and PSA-NCAM expression in the subventricular zone of adult rats. AB - The subventricular zone (SVZ) bordering the lateral ventricle is one of the few regions of adult brain that contains dividing cells. These cells can differentiate into neurons in vivo after migration into the olfactory bulb and in vitro in the presence of appropriate growth factors. Little is known, however, about the fate of these cells in vivo after brain injury in adults. We examined cell number and expression of differentiation markers in the SVZ of adult rats after cortical lesions. Aspiration lesions of the sensorimotor cortex in adult rats induced a transient doubling of the number of cells in the SVZ at the level of the striatum without consistent increases in bromodeoxyuridine-labeled cells. Immunoreactivity to the polysialylated neural cell adhesion molecule, expressed by the majority of cells of the SVZ during development, increased dramatically after lesion. In contrast, immunolabeling for molecules found in mature neurons and glia did not increase in the SVZ after lesion, and immunoreactivity for growth factors that induce differentiation of SVZ cells in vitro decreased or remained undetectable, suggesting that lack of appropriate growth factor expression may contribute to the lack of differentiation of the newly accumulated cells in vivo. The data reveal that cells of the SVZ are capable of plasticity in the adult rat after brain injury in vivo and that the newly accumulated cells retain characteristics seen during development. PMID- 8725351 TI - Concentration and storage of biotin in the amphibian brain. AB - Prominent displays of endogenous biotin reactivity can be observed at specific locations in histochemical preparations of the forebrain and midbrain in the northern leopard frog (Rana pipiens) and common American toad (Bufo americanus). At the light microscopic level, the biotin reactivity appears in clusters of darkly stained puncta of either spherical or rodlike shape in the olfactory cortex, nucleus isthmi, and hypothalamus. With the electron microscope, the biotin reactive spheres are identified as neuronal varicosities and synaptic boutons and the rods as short segments of axons. Appropriate controls demonstrate that the punctate biotin-reactive structures are sites of concentration of biotin or a biotin analog in the processes of certain neurons. These data represent the first observation on the selective concentration of a vitamin in vertebrate neurons and suggest that biotin may have specialized functions in anatomically delimited areas of the central nervous system. Localization of the densest concentration of the biotin-reactive puncta in the dorsolateral prominence of the olfactory cortex may have relevance to the functional organization of the olfactory system. The distributions of biotin-reactive puncta were observed in laboratory-housed frogs and in wild toads captured in the summer months but were sparse or absent in batches of commercially obtained frogs examined immediately upon arrival in the laboratory. Systemic administration of biotin or biocytin hydrochloride did not alter the appearance or numbers of the biotin-reactive structures either in newly received or laboratory-housed frogs. These findings suggest that the capacity of the biotin-storage mechanism in the amphibian brain may be set by environmental factors and may be readily saturable from natural dietary or enteric sources. PMID- 8725352 TI - [Current status of adhesive resin restorations]. PMID- 8725353 TI - [Suppression of granulopoiesis by vesnarinone]. AB - The effects of vesnarinone (3,4-dihydro-6-[4(3,4-dimethoxybenzoyl)-1-piperanizyl] 2 (1H)-quinolinone) on the hematopoietic precursors in 5 healthy volunteers and leukemic blast progenitors in 11 acute myeloid leukemia (AML) patients, 1 chronic myelocytic leukemia patient (CML) in blast crisis, and 3 leukemic cell lines were studied in methylcellulose and suspension cultures. Normal erythroid precursors (colony-forming unit erythroid: CFU-E and burst-forming unit erythroid: BFU-E) and granulopoietic precursors (colony-forming unit granulocyte/macrophage: CFU GM) were suppressed in methylcellulose culture by vesnarinone in a dose-dependent manner. Leukemic blast progenitors may replicate themselves and/or undergo terminal divisions with limited differentiation. The plating efficiency of primary blast colony formation (PE1) in methylcellulose, which is considered to reflect the terminal divisions of leukemic blast progenitors, was suppressed by vesnarinone in a dose-dependent manner in all cases tested. The plating efficiency of secondary blast colony-formation (PE2) in methylcellulose culture and the recovery of clonogenic cells in the suspension culture, which are considered to reflect the self-replication function of leukemic blast progenitors, were also suppressed by vesnarinone in a dose-dependent manner in all cases tested. The results suggest that vesnarinone inhibits the growth of normal and leukemic hematopoietic progenitors. To determine the mechanism by which vesnarinone inhibits hematopoiesis, the effect of the agent on apoptosis (programmed cell death) of leukemic cells was studied. DNA ladder formation was recognized in OCI/AML 1 a cells after exposure to 100 micrograms/ml vesnarinone for 18 hours; this means that vesnarinone induced apoptosis in this cell line. Therefore, vesnarinone is considered to be the cause of apoptosis of granulopoietic precursors. PMID- 8725354 TI - [An electromyographic study on functional adaptations--associated with herbst appliance]. AB - The purpose of this study was to examine functional muscular adaptation to changes in sagittal jaw relation by electromyography. The structural changes were induced by the Herbst appliance. The subjects consisted of four patients with Angle Class II Division 1 malocclusion. The structural changes were evaluated by lateral cephalograms. Functional muscular adaptation was evaluated by electromyogram (EMG) of masticatory muscles, including the inferior head of the lateral pterygoid (ILpt) muscle. The EMG of the ILpt muscle was recorded by using an intraoral surface electrode. The same pattern of cephalometric changes was observed in all patients. The ILpt muscle activity increased immediately after wearing the appliance just like when the construction bite position was taken. However, it reduced remarkably after 4 to 6 months in all patients. On the other hand, the superimposed cephalometric tracings indicated that the locations of the condyles related to the glenoid fossae were still more anteroinferior than their original positions 4 to 6 months after wearing the appliances in three patients. These findings seem to indicate multifactorial effects on the adaptation of the muscle function. It was concluded that functional adaptation to the newly-created morphological changes were not dependent only on the intensity and the duration of the functional stimulation. PMID- 8725355 TI - [Continuous evaluation of muscle activity with occlusal tooth contacts part II. Nocturnal tooth contacts and muscle activity]. AB - In order to evaluate the relationship between the mandibular position and the muscle activity during bruxism, an originally-designed device for detecting tooth contacts in eccentric jaw positions in addition to the intercuspal position was applied to 7 subjects and electromyography was carried out stimultaneously. Nocturnal bruxing patterns such as lateral grinding, clenching in the intercuspal position or eccentric jaw position were clearly distinguished by using this system. In one of 3 subjects who were not aware of bruxism, both the duration and the frequency of the muscle activity were significantly higher than those of other subjects. This suggests that some persons with bruxism who are unaware of it actually grind or clench their teeth during sleep. Tooth contacts during sleep were classified into 6 patterns according to jaw position. In the IP pattern, both regular and irregular EMG patterns were observed without any lateral movement of the mandible. Furthermore, each subject had the mandibular position on the preference side during tooth contacts and the EMG activity of the anterior temporal muscle on the preference side during the IP pattern was higher than that on the opposite side. This system enabled detailed evaluation of bruxism. PMID- 8725356 TI - [Functional analysis of transforming growth factor-beta type I receptor]. AB - Proteins in the transforming growth factor-beta (TGF-beta) superfamily exert their effects by forming heteromeric complexes of their type I and type II serine/threonine kinase receptors. Binding of TGF-beta to its constitutively active type II receptor (T beta R-II) recruits the type I receptor (T beta R-I) into the complex; T beta R-I is thereafter phosphorylated in GS domain located just N-terminal to the kinase domain and activated, which is necessary for TGF beta signaling. Here we investigated the role of cytoplasmic juxtamembrane region located between the transmembrane domain and the GS domain of T beta R-I by mutational analyses using mutant lung epithelial cells which lack endogenous T beta R-I. Upon transfection, wild-type T beta R-I restored the TGF-beta signals for growth inhibition and production of plasminogen activator inhibitor 1 (PAI)-1 and fibronectin. A deletion-mutant, T beta R-I/JD 1 (delta 150-181), which lacks the juxtamembrane region preceding the GS domain bound TGF-beta in concert with T beta R-II and transduced a signal leading to production of PAI-1 but not growth inhibition. Recombinant receptors with mutations that change serine172 to alanine (S172A) or threonine176 to valine (T176V) were similar to wild-type T beta R-I in their abilities to bind TGF-beta, formed complexes with T beta R-II, and transduced a signal for PAI-1 and fibronectin. Similar to T beta R-I/JD1 (delta 150-181), however, these missense mutant receptors were impaired to mediate a growth inhibitory signal. These observations indicate that serine172 and threonine176 of T beta R-I are dispensable for extracellular matrix protein production but essential to the growth inhibition by TGF-beta. PMID- 8725357 TI - [Analysis of a genetic factor of metal allergy--polymorphism of HLA-DR, -DQ gene]. AB - To reveal immunogenetic factors involved in the pathogenesis of metal allergy, 30 unrelated Japanese patients with histories of metal allergy were typed for HLA. They were confirmed by an unequivocal positive patch-test reaction to mercury, nickel or palladium. The HLA-DR and -DQ DNA were typed by using the polymerase chain reaction-sequence specific oligonucleotide probe (PCR-SSO) method. The frequency of each allele in the patient groups was compared to that in normal Japanese subjects. In mercury sensitive patients, there was no significant HLA association. In nickel sensitive patients, there was a significant increase of DRB4 (p < 0.05) but no significant association of DRB1 nor DQ locus, although there was an increase of DRB1*0405 (R.R = 2.36). In palladium sensitive patients, there were significant increases of DRB4 (R.R. = 15.48, p < 0.05), DR4(R.R. = 13.27, p < 0.005), DRB1*0405(R.R. = 4.43, p < 0.05), and DQB1*0401 (R.R. = 4.59, p < 0.05), and significant decreases of DRB5 (R.R. = 0.09, p < 0.05), and DQA1*0103 (R.R. = 0, p < 0.05). PMID- 8725358 TI - [Effects of lateral mandibular deviation on masseter muscle activity]. AB - The investigation of the physiological response of masticatory muscle which is associated with the perception of the mandibular lateral deviation is clinically important. Therefore, the effects of various lateral mandibular deviation on the masseter muscle function were investigated on five healthy subjects. The mandibular position was deviated 0.5, 1.0, 1.5, 2.0 and 3.0 mm to the right and left from a reference position corresponding to the rest position. The mandible was passively retained in each test position by means of a bite block. Electromyographic activity (background activity) was recorded with bipolar surface electrodes applied to the right and left masseter muscles. Jaw-jerk reflex was evoked by a reflex hammer, and motoneuron activity was indirectly examined. The background activity as well as the amplitude of the jaw-jerk reflex on the mediotrusive side mainly increased in proportion to the mandibular deviation. However, most subjects showed no significant change about 3 mm from the reference position in the background activity and about 2 mm in the range of muscle activity evoked by the jaw-jerk reflex. These results suggested that the masseter jaw-jerk reflex may assist clinical examination of small mandibular deviations. PMID- 8725359 TI - [Influence of various impression procedures for mandibular distal extension removable partial denture on displacement of residual ridge]. AB - In order to investigate the influence of various impression procedures on the morphology of the partially edentulous mucosal tissue precisely, a new 3-D measurement system based on a binocular-temporal gradient method was devised. The greatest advantage of the system is that the amounts of substance and minute deformation of the object can be easily measured simultaneously. The accuracy of the system is 0.1mm and 10mm in the substance measurement and in the deformation measurement respectively. Three parameters selected for the impression procedure were the space between the inner surface of the tray and the mucosal tissue, the flow of impression material and the size of tray. It was found that the amount of displacement in the mucosal tissue was increased as the thickness of space on the tray was reduced or the viscosity of the impression material was increased in partially edentulous patients. Moreover, the large tray caused greater displacement around the margin than the small one. PMID- 8725360 TI - [Neurophysiological mechanism of jaw-tongue reflex in man]. AB - The present study was carried out to examine if the jaw-tongue reflex (JTR) could be elicited in man and if afferent fibers from the jaw-closing muscles were involved in this reflex. Electromyographic (EMG) activities were simultaneously recorded from the genioglossus (GG), the tongue retruding (TR) muscles, and the anterior temporal (TA), the posterior temporal (TP), the masseteric (Mass) and the digastric (Dig) muscles under various conditions. The results were as follows: 1. EMG activities of the GG and the TR muscles were increased during clenching, voluntary and passive jaw-opening, and chin-tapping. 2. Electrical stimulation of the deep temporal and masseteric nerves evoked H-reflex in the TA and the Mass muscles. EMG activities in the GG and the TR muscles were detected as the intensity of the electrical stimulation were increased. 3. The tonic vibration reflex (TVR) was induced in the jaw-closing muscles by applying vibratory stimulation to the bellies of the TA and the Mass muscles. EMG activities of the GG and the TR muscles were increased as the TA muscle activity was increased. It is concluded that the JTR is elicited in man and suggested that low threshold afferent fibers from the jaw-closing muscles, mainly the temporal muscle, were involved in eliciting the JTR in man. PMID- 8725361 TI - [Immunohistochemical localization of Type I, II and IX collagens in pleomorphic adenoma of human salivary glands]. AB - The expression patterns of type I, II and IX collagens in chondromyxoid tissue of salivary pleomorphic adenomas were examined by immunohistochemistry. In the early stage of cartilage development, type IX collagen was detected intracytoplasmically, mainly in the proliferating myoepithelial cells and not in the extracellular matrix. Proliferating myoepithelial cells did not show chondrocytic characteristics at this stage. Type I and II collagens were co distributed in the extracellular matrix of myxoid tissue. In the chondroid tissue, variable immunostaining patterns of type I and II collagens were also observed. Some proliferating myoepithelial cells in myxoid and chondroid tissue were immunostained with anti-alpha-smooth muscle actin antibody indicating their myoepithelial origin. These results suggested that proliferating myoepithelial cells may be the origin of the chondroid tissue and the expression of type IX collagen in cartilaginous tissue precedes the expression of type I and II collagens. PMID- 8725362 TI - [Influences of orthognathic surgery to facial skin sensation--an analysis with a new testing method]. AB - This study introduces an original testing method on facial skin sensation and describes the results obtained by its application to patients after orthognathic surgery. The alterations of skin sensation in lower lip and chin area were examined in 135 patients who had undergone orthognathic surgery, by application of electrical stimulation through an original pencil shaped probe. Detection threshold of the same area before surgery were used as control, and the location and size of the area with thresholds higher than control levels were measured. The control thresholds ranged from 0.13 mA to 0.21 mA. The higher threshold area was found in 86.3% of all the patients in 1 month after surgery, in 27.7% after 1 year. The threshold levels declined with the time, and in none of the patients the threshold level was higher than 1.0 mA more than 2 years after surgery. The patients whose threshold level was more than 1.0 mA in 1 month postoperatively showed a tendency to have the area with a threshold higher than the control level even 1 year later (p < 0.0001). The threshold of the younger patients tended to improve smoothly than the older. In addition, the patients with a shorter set back or advance distance had a slighter sensory disturbance and recovered more smoothly than those with a longer distance. PMID- 8725363 TI - [Depression of reciprocal Ia inhibition of crural motoneurons during rhythmical jaw movement in rabbit]. AB - The purpose of this study was to investigate how the reciprocal inhibition of the ankle-flexor motoneuron by the ankle-extensor spindle afferents is altered during cortically-induced rhythmical jaw movement (RJM) in urethane-anesthetized rabbit. The monosynaptic reflex (MSR) was induced by test shock of the common peroneal nerve (CPN) and recorded from a distal point of the nerve. The conditioning shock applied to the tibial nerve (TN) at the intensity below 1.2 times the threshold caused a significant inhibition of the MSR in the CPN. The inhibition tended to slightly decrease with an increase in amplitude of the test MSR with either application of stronger test shock or during RJM. The decrease of the reciprocal inhibition significantly exceeded the degree that can be ascribed to the increase of the amplitude of the MSR during RJM. The reciprocal inhibition was significantly reduced during fictive as well as actual RJM. It was concluded that (1) the reciprocal Ia inhibition of the crural motoneurons in the rabbit is reduced with RJM and (2) the intraoral sensory input is not essential for this reduction. This study indicated that the oral motor activity generally exerts a strong influence on the bodily motor function. PMID- 8725364 TI - [Effects of craniofacial pain on jaw jerk in man]. AB - Jaw jerk was evoked in 12 normal subjects (N group) and 12 patients with unilateral craniofacial pain who had neither mandibular deviation nor organic changes in the temporomandibular joint (P group), about every ca. 2 sec by tapping the center of the chin with a reflex hammer, in which a microswitch was embedded to indicate the time of its contact with the chin. The surface electromyogram of the masseter muscle was recorded bilaterally. It was found that (1) in the N group, there was no difference in the onset latency between the right (6.30 +/- 1.06 ms; n = 192) and left (6.32 +/- 1.04 ms; n = 192) sides (P > 0.05); (2) in the P group, the latency was shorter on the painful side than on the pain-free side; and (3) after pain was relieved by treatment, the bilateral difference in the latency of jaw jerk disappeared, and the latencies fell to that in normal subjects. It was concluded that the bilateral difference of the latency of jaw jerk can be used as an investigative criterion for the existence of pain. PMID- 8725365 TI - [Purification of serotype d-- and e--specific antigens from Actinobacillus actinomycetemcomitans and seroclassification of clinical isolates from periodontal patients]. AB - Oral Actinobacillus actinomycetemcomitans strains have been classified into five serotypes. The aim of this study was to determine the compositions of A. actinomycetemcomitans serotype d- and e- specific antigens. The serodistribution of clinical isolates from the patients with periodontitis were also investigated. Serotype-specific polysaccharide antigens of A. actinomycetemcomitans IDH 781 (serotype d) and OMZ 534 (serotype e) were extracted from whole cells by autoclaving. The extracts were purified by chromatography on DEAE-Sepharose CL-6B and Sephacryl S-300HR columns. The serotype d antigen was composed of rhamnose (17.1%), mannose (45.5%), galactose (2.0%) and glucose (35.5%). On the other hand, the serotype e antigen was composed of rhamnose (23.9%), mannose (29.1%), galactose (11.0%), glucose (13.5%) and unidentified sugar (22.5%). Immunodiffusion tests revealed that the purified polysaccharide antigen form a single precipitin line with the corresponding rabbit antiserum. A total of 157 A. actinomycetemcomitans clinical isolates from diseased sites of 39 patients with periodontitis were serotyped by using serotype-specific rabbit antisera against A. actinomycetemcomitans serotype a, b, c, d and e strains. In the immunodiffusion assay, the autoclaved extracts of 42, 12, 34, 8 and 41 A. actinomycetemcomitans clinical isolates reacted with serotype a, b, c, d and e antisera, respectively. These findings indicate that the extraction of serotype antigens by autoclaving is useful and definite for the serotyping of A. actinomycetemcomitans clinical isolates. PMID- 8725366 TI - [A study on colorimetry of oral mucosal lesions]. AB - Color observation is essential to diagnose oral mucosal lesions. In order to measure and record the color of the oral mucosa more objectively, colorimeters were evaluated. The noncontact-type colorimeter showed considered values comparable with those of visual color matching and was reliable. The color distribution of the normal oral mucosa was from 5.0R to 4.1YR in hue, 3.5 to 6.0 in value, and 3.7 to 6.7 in chroma. The color distribution of leukoplakia was from 2.6RP to 3.4YR in hue, 3.7 to 7.0 in value, and 0.3 to 8.3 in chroma and the ranges thereof were considerably broader than those of normal mucosa and other mucosal lesions. The higher chroma of leukoplakia was, the severer epithelial dysplasia was. The color distribution of oral cancer was from 3.0R to 8.2YR in hue, 2.7 to 6.4 in value, and 3.0 to 7.8 in chroma. The ranges of color distribution was broader than those of normal mucosa and narrower than those of leukoplakia. The color distribution of lichen planus was from 1.7R to 8.9R in hue, 3.5 to 6.3 in value, and 3.6 to 8.5 in chroma. The chroma of erythema is the highest of all mucosal lesions. Some oral mucosal lesions are suggested to have characteristic colors. PMID- 8725367 TI - [Histological study on experimental tooth movement in alveolar bone defects implanted with decalcified bone powder]. AB - Recently, secondary bone grafting for the alveolar cleft of cleft lip and palate patients has carried out generally with the object of connecting the alveolar cleft, of leading the tooth eruption, of control the tooth axis. In this study, the experimental tooth movements to the area were tested and the histological changes were investigated with adult cats. The experimental defects of 2mm in diameter were made at the alveolar bone mesial to the both maxillary first molar. At the experimental side, DBP were stuffed into the defect. Only defects were made at the control side. The tooth were moved for 6 weeks. The restoration of the defect hadn't occurred at the control side after 14 weeks. While appositional bone growth and new bone formation surrounding the DBP were observed, and complete restoration had be already occurred at experimental side after only 8 weeks. In the group which tooth movement were started after 1 week, the existing alveolar bone were bent to mesial at both side. In the group were started after 8 weeks, the existing alveolar bone were bent to mesial at the control side, either. The DBP and new bone were resorbed at the same time at the experimental side. PMID- 8725368 TI - [Tooth movement by biomechanical technique in orthodontic treatment]. AB - The purpose of this study was to analyze tooth movement by measuring the electric impedance of the wire and by using the nonlinear finite element analysis. In this study, the HP4338A Milliohmmeter was used and the model consisted of a metal tooth, silicone bath, bracket, and orthodontic wire. Then, the two-dimensional finite elements were used and the model consisted of tooth, periodontal ligament, alveolar bone, bracket, and orthodontic wire. There were two analyses. The first was the contact nonlinear finite element analysis of elastic deformation in the displaced tooth at the slide points between the bracket and wire where two gap element areas existed, and the second was the material nonlinear finite element analysis of the stress relaxation in bone at the fixing points between the bracket and wire where two rigid element areas, differing from the gap element areas, existed. The results were as follows: 1. The linear proportion between the electric impedance and the orthodontic wire was found. Therefore very minute tooth movements became measurable with changes of the wire electric impedance. 2. During tooth movement, the stick between the bracket and the wire altered with the slip ones repeatedly. 3. The movement of the displaced tooth at the center of rotation was counterclockwise. 4. The movement of the displaced tooth at the bracket position was clockwise. PMID- 8725369 TI - [Effects of smeared layer on luting strength of dental cements]. AB - Effects of smeared layer on the bond strength between dentin and dental cements were evaluated. Four surface conditions (No treatment, K-etchant, Dentin conditioner and EDTA) were prepared on the ground dentin of bovine frontal teeth. A copper tube was fixed and filled with a dental cement (zinc-phosphate cement or glass-polyalkenoate cement). A hook was embedded in the tube, with or without 1 MPa load during cementation. The bond strength was measured by a tensile test using a universal testing machine. No treatment had the largest bond strength of zinc-phosphate cement with or without load. No treatment, Dentin conditioner and EDTA had large bond strengths of glass-polyalkenoate cement with or without load; the bond strength of K-etchant was small without load and comparable to those of other surface conditions when load was applied. These results suggested that the bond strength of three surface conditions was not stronger than that of no treatment. Therefore, the elimination of smeared layer was concluded to have no advantage for increasing the bond strength between dentin and dental cements. PMID- 8725370 TI - Hepatocellular carcinoma: comparative study among precontrast, artery-dominant, and delayed-contrast phase images at ultrafast CT. AB - To evaluate the detectability of hepatocellular carcinoma (HCC) at artery dominant phase and to compare it with that at precontrast and delayed-contrast phases, 34 patients with 109 HCCs were examined by ultrafast computed tomography (CT) using an incremental dynamic technique under single breath-holding. Images of three phases (precontrast, artery-dominant, and delayed) were compared with iodized-oil CT images. None of the lesions were detected only at precontrast phase. HCC detectability at the artery-dominant phase (87/109, 80%) was significantly superior to that at precontrast phase (32/109, 29%) (p < .01) and delayed-contrast phase (57/109, 52%) (p < .01). All 31 lesions missed at delayed contrast phase were depicted at artery-dominant phase, while one lesion observed at delayed-contrast phase was missed at artery-dominant phase. Incremental dynamic CT at artery-dominant phase was suitable for the diagnosis of HCCs. Precontrast phase images were not necessary, although delayed-phase images were useful for the evaluation of tumor extension in some cases. PMID- 8725371 TI - Assessment of treatment response in irradiated lung cancer by sequential thallium 201 SPECT: comparison with tumor volume change and survival time. AB - We carried out a study to evaluate treatment response and residual mass in irradiated lung cancer with single photon emission computed tomography (SPECT) using 201TI, and investigated the relation among 201TI uptake ratio, tumor volume, and patient survival time. 201TI uptake ratios and tumor volume measured before and after radiotherapy were compared in 14 patients. We divided them into a complete response (CR) group (n = 5) and partial response + no change (PR + NC) group (n = 9) according to the tumor volume response determined by CT, and a longer survival (LS) group (n = 7) and shorter survival (SS) group (n = 7) according to the survival time. The CR group showed smaller 201TI uptake ratios after irradiation (5.3 +/- 6.0%) than the PR + NC group (52.0 +/- 27.4%) (p < 0.01), but there were no significant differences in 201TI uptake ratio before irradiation or survival time in the other two groups. The LS group showed a smaller 201TI uptake ratio before (107.8 +/- 48.3%) and after (17.0 +/- 14.4%) irradiation than the SS group (126.6 +/- 34.4% p < 0.01, 52.6 +/- 27.5% p < 0.05), but there were no significant differences in tumor volume before or after irradiation in the other two groups. We concluded that 201TI SPECT is very useful to evaluate treatment response in irradiated lung cancer and can play a supplementary role in evaluating prognosis. PMID- 8725372 TI - Doppler spectral analysis of blood flow velocities in common femoral artery: age related changes in healthy subjects and characteristics of abnormal hemodynamics in obstructive arterial disease. AB - Flow velocity patterns in the common femoral artery were analyzed in 66 lower extremities of 33 healthy subjects and 18 lower extremities of 12 patients with arteriosclerosis obliterans (ASO) by Doppler ultra-sonography. In a phantom study, Doppler flow velocity showed a close correlation with actual flow velocity: y = 1.02x - 1.22 (r = 0.995). In clinical trials, 33 healthy subjects were classified into three groups: group I (20-40 years), group II (41-60 years) and group III (61-80 years). Starting time of acceleration (STA) was shortened as age increased. Peak reverse velocity (PRV), maximum deceleration rate (MDR), and pulsatility index (PI) were significantly decreased in group III as compared with groups I and II. In patients with ASO, STA was prolonged, and PFV, PRV, MAR, MDR, PI, VSA, and FV were significantly decreased (p < 0.001) as compared with healthy control group III. Consequently, marked characteristic changes such as flattening of curves and disappearance of reverse flow in Doppler flow wave-form were observed. These data obtained by Doppler ultrasonography are thought to be useful for the early diagnosis and follow-up of patients with ASO. PMID- 8725373 TI - Decreased signal intensity of cerebral cortex on T2-weighted MR images. AB - To define the frequency of decreased signal intensity (DSI) in cerebral cortex on T2-weighted images relative to aging and to the incidence of identifying white matter pathology, T2-weighted MR brain images of 906 patients consecutively examined between July 1989 and June 1991 were reviewed. MR images of cerebral cortex were divided into five areas: frontal lobe (F), pre- and postcentral gyri (C), parietal lobe (P), occipital lobe (O) and temporal lobe (T). Each area was separately and independently evaluated for the presence or absence of DSI. The frequency of DSI in each area was plotted against patients' ages. The severity of leukoaraiosis was evaluated in relation with DSI in each cortical area. Patients with DSI in area F numbered 61 (5.7%), in area C 236 (26.0%), in area P 174 (15.9%), in area O 428 (47.2%), and in area T 10 (1.1%). The number of patients with DSI in each area increased with age. The greater the severity of leukoaraiosis, the greater the chance of finding DSI of the cerebral cortex. The incidence of DSI in the cerebral cortex increased with the patient's age as well as with the severity of co-existent white matter pathology. PMID- 8725374 TI - MR imaging of lingual carcinoma: comparison with surgical staging. AB - The purpose of this study was to determine the usefulness of magnetic resonance (MR) imaging for the preoperative staging of 18 patients with lingual carcinomas. Tumor stage as determined by MR imaging was compared with pathological stage. Conspicuity of tumors was compared among dynamic MR, T2-weighted, and postcontrast T1-weighted images. The tumor stage was evaluated correctly with MR imaging in 15/18 patients (83.3%). One patient was overstaged and two understaged due to incorrect diagnosis of size. In all T4 cases, tumor extension was diagnosed correctly. Dynamic MR and T2-weighted images were superior to postcontrast T1-weighted images in delineating and showing the extension of carcinomas. However, dynamic MR imaging showed no significant superiority to T2 weighted imaging. We conclude that MR imaging is of great value in the staging of lingual carcinoma. Dynamic study should be performed only when the lesion is undetectable or equivocal on T2-weighted imaging. PMID- 8725375 TI - Gallium-67 scintigraphy in evaluation of malignant lymphoma of the thyroid gland. AB - During the period from 1982 to 1994, a total of 45 67Ga scintigrams were performed in 12 patients with malignant lymphoma of the thyroid gland. Eight scintigrams were performed before treatment, 12 during the course of radiation therapy, and 25 after treatment. Scintigrams before treatment showed intense uptake in the thyroid masses irrespective of their size. Two of four scintigrams revealed no accumulation in the clinically remaining masses that were given a radiation dose of less than 20 Gy. No accumulation or thyroid masses were observed in cervical regions that received an irradiation dose of 20 Gy or more. Distant involvement in the upper half of the body could be detected by 67Ga scintigraphy in all cases, while there were two false negative lesions in the abdominal or pelvic cavity. Abdominopelvic lesions involving the gastrointestinal tract were observed in four cases, including two autopsied cases. 67Ga scintigraphy is useful for the evaluation of malignant lymphoma of the thyroid gland, but other examinations are recommended when 67Ga uptake is observed in the abdominal or pelvic cavity. PMID- 8725376 TI - Gastric carcinoma metastasis to calf muscles: MR findings. AB - We report the MR findings of a rare case of gastric carcinoma metastasis to muscles in the calf. T2-weighted images showed the diffuse enlargement of skeletal muscles in the calf with remarkable hyperintensity. PMID- 8725377 TI - Myelolipoma of the adrenal gland found incidentally during abdominal ultrasound and computed tomography examinations. AB - We report a case of adrenal myelolipoma found incidentally during abdominal ultrasound (US) and computed tomography (CT) examinations for hypertension. The patient was a 34-year-old woman with a history of hypertension. In June 1992, she complained of dizziness and easy fatigability and went to a hospital, where she was diagnosed as very hypertensive. She was hospitalized for further examinations. Abdominal US and CT incidentally revealed a right adrenal tumor. The tumor was visualized as a hyperechoic mass on US and an inhomogeneous low density mass with inner fat density and focal calcification on CT. Magnetic resonance (MR) imaging visualized the tumor as a high signal intensity mass on both T1- and T2-weighted images. Since the tumor was relatively large and the possibility of malignancy could not be completely excluded, resection of the tumor was performed. The resected tumor, which measured 5.2 x 3.8 x 2.5 cm, was located in the right adrenal medulla and was composed of mature fat cells and bone marrow elements, with all the developmental stages of hematopoietic cells. Small areas of fresh hemorrhage and calcification were found. The fat cell component showed focal myxoid change, but there were no immature or atypical cells. These features were consistent with myelolipoma. The clinical and pathologic features of adrenal myelolipoma are discussed. PMID- 8725378 TI - Familial glioma in two siblings. AB - We report two siblings who had intracranial gliomas, one with glioblastoma multiforme and the other with mixed oligo-astrocytoma. The genetic influences in the development of glioma are discussed. PMID- 8725379 TI - Rotational digital angiography of ulcer-like projection of pelvis. AB - Ulcer-like projection is an angiographic sign of thrombosed aortic dissection. However, no pelvic manifestation of ulcer-like projection of aortic dissection has been reported. We report three patients in whom ulcer-like projections of the pelvic artery were revealed by rotational digital angiography using the newly developed computerized dynamic stereo-radiography apparatus. PMID- 8725380 TI - A personal view of pharmacology. AB - This essay is an account of the author's experience in trying to interpret the action of drugs as seen in in vitro bioassays. The central theme is how the development of simple mathematical models has assisted in the interpretation of drug actions. Starting from encounters with partial agonists, the essay describes the development of an operational model of agonist activity, the significance of receptor distribution, the analysis of indirect competitive antagonism and various two-receptor systems, followed by examples of pharmacological resultant activity. Analyses of tissue and species variations are described. The essay ends with an assessment of the future of bioassay in pharmacology. PMID- 8725381 TI - A career in toxicology. PMID- 8725383 TI - Oxidative stress in neurodegenerative diseases. AB - Oxidative stress refers to the cytopathologic consequences of a mismatch between the production of free radicals and the ability of the cell to defend against them. Growing data from experimental models and human brain studies suggest oxidative stress may play an important role in neuronal degeneration in diseases such as Parkinson's disease, Alzheimer's disease, and amyotrophic lateral sclerosis. Mitochondrial oxidative metabolism, nitric oxide, phospholipid metabolism, and proteolytic pathways are potential sources of intracellular free radicals. Alterations in free radical defense systems may also contribute to oxidative stress. A net increase in reactive oxygen species can produce damage to lipids, proteins, and DNA and induce necrosis or apoptosis. Elucidating the pathways important in the production of and defense from free radicals may be important in devising new pharmacologic strategies to slow or halt neuronal degeneration. PMID- 8725382 TI - Antiprogestins: mechanism of action and contraceptive potential. AB - Antiprogestins are characterized by substitutions at the 11 beta and 17 alpha positions of the steroid ring system and bind strongly to both progesterone and glucocorticoid receptors. Although they function predominantly as antiprogestins and antiglucocorticoids, on occasion they display progestin agonistic and even antiestrogenic properties. The most common clinical use of the antiprogestin mifepristone is to induce a medical abortion in the early stages of pregnancy. Progesterone maintains the endometrium, transforming it from a proliferative to a secretory state. It also facilitates the luteinizing hormone surge, which initiates ovulation. As a consequence, antiprogestins may also have contraceptive potential. Although antiprogestins do delay ovulation, this effect is inconsistent unless high doses are given, and under these circumstances, the antiprogestin effect is associated with unopposed estrogen action on the endometrium. Very low doses of antiprogestins do not affect hormonal secretion or ovulation or alter bleeding patterns, but they do have contraceptive potential by inducing profound alterations in endometrial morphology. Mifepristone is also a very effective and safe postcoital agent. This new class of pharmacological agents has numerous other gynecological and obstetrical indications, such as endometriosis, uterine myoma, and expulsion of the fetus in the case of fetal death in utero. Antiprogestins may also be used in the treatment of steroid dependent tumors. There are also therapeutic implications consequent to their antiglucocorticoid properties. PMID- 8725384 TI - Progress in antisense oligonucleotide therapeutics. AB - The past several years have seen substantial progress in the development of antisense oligonucleotides as pharmacological tools and as therapeutic agents. With properly designed and executed experiments, it has been possible to demonstrate selective inhibition of gene expression, owing to an antisense mechanisms of action both in cell culture-based experiments and in vivo. As the field has matured, it has also realized that oligonucleotides can produce a variety of effects in cell culture and in vivo that cannot be ascribed to an antisense mechanism of action. Nevertheless, with proper controls it has been possible to demonstrate that the pharmacological activity of an oligonucleotide is consistent with an antisense mechanism of action. The pharmacokinetic properties of first-generation phosphorothioate oligodeoxynucleotides and their toxicological limitations have been characterized in rodents, primates, and humans. Finally, it has been demonstrated that medicinal chemistry can improve the properties of oligonucleotides, as several modifications have been identified that have increased potency, altered pharmacokinetic properties and potentially decreased toxicological liabilities. PMID- 8725385 TI - Molecular mechanisms of toxicant-induced immunosuppression: role of second messengers. AB - Changes in immunocompetence following chemical exposure have been established for a wide variety of unrelated agents. For the vast majority of immunotoxic compounds thus far identified, disruption of normal immune function is clearly mediated through direct interactions between the agent, or its metabolite, and immunocompetent cells. Regardless of whether this interaction occurs at the level of the cell membrane or at intracellular sites, basic regulatory processes mediated by second messengers are often altered. These alterations can ultimately result in immunologic dysfunction, which is most often manifested as immunosuppression. The specific disruptions in intracellular signaling produced by a number of immunotoxic compounds have now been identified, leading to a basic understanding of their molecular mechanism of action. Equally important, through the application of these agents as biological probes, new insights have been gained pertaining to which intracellular processes control which cellular functions within various populations of immunocompetent cells. PMID- 8725386 TI - P-glycoproteins and multidrug resistance. AB - Multidrug resistance represents a major obstacle in the successful therapy of neoplastic diseases. Studies have demonstrated that this form of drug resistance occurs both in cultured tumor cell lines as well as in human cancers. P glycoprotein appears to play an important role in such cells by acting as an energy-dependent efflux pump to remove various natural product drugs from the cell before they have a chance to exert their cytotoxic effects. Expression of the MDR1 gene product has been associated with a poor prognosis in clinical studies. It has been demonstrated in the laboratory that resistance mediated by the P-glycoprotein may be modulated by a wide variety of compounds. These compounds, which include verapamil and cyclosporin, generally have little or no effect by themselves on the tumor cells, but when used in conjunction with antineoplastic agents, they decrease, and in some instances eliminate, drug resistance. Clinical trials to modulate P-glycoprotein activity are underway at the present time to determine if such strategies will be feasible. Although the P glycoprotein is expressed in many cell lines and occurs in patient tumors, its expression is not a universal feature of multidrug resistance, suggesting that other mechanisms are operating. PMID- 8725387 TI - Lipid vs protein theories of alcohol action in the nervous system. AB - There has been a long-standing debate concerning whether alcohols produce their effects in the central nervous system (CNS) by acting on lipids or proteins. Lipid theories postulate that alcohols act via some perturbation of the membrane lipids of CNS neurons, whereas protein theories propose that alcohols act by interacting with a neuronal protein site. Although the primary site of action differs in the two theories, both theories postulate that the CNS effects of alcohols ultimately result from alterations in protein function. This review discusses lipid and protein theories of alcohol action and the evidence supporting these theories. In addition, the effects of alcohols on the function of neurotransmitter-gated ion channels are discussed, as several types of these receptor-ion channels have been found to be sensitive to the actions of alcohols, and recent studies on those actions have yielded new insights into the question of whether the primary action of alcohols involves lipids or proteins. PMID- 8725388 TI - Molecular mechanisms of estrogen carcinogenesis. AB - In western society, the causes of several cancers--including breast, endometrium, ovary, liver, and prostate--have been linked to inappropriate and/or prolonged exposure to synthetic or endogenous steroidal hormones. In this review, we discuss the mechanisms of estrogen carcinogenesis with a focus on estrogen metabolism to 16 alpha-hydroxy estrone and 2- and 4-hydroxy catechol estrogens and the potential effects of these metabolites in vitro and in vivo on hamster liver and kidney and rat liver carcinogenesis models. The examples demonstrate that the parent compounds and their metabolites cause both nongenotoxic cell proliferative effects as well as direct and indirect genotoxic effects, which illustrates the complex nature of estrogen carcinogenesis. These effects, in combination with the metabolic state of the tissue and the timing of its exposure, may determine the cell type (organ) of tumor development and the severity of disease. PMID- 8725389 TI - Cardiac actions of antihistamines. AB - Two of the nonsedating antihistaminic drugs, terfenadine and astemizole, have recently been recognized in rare cases to induce the syndrome of torsades de pointes, i.e. QT interval prolongation and life-threatening ventricular tachycardia. Each was found to prolong cardiac repolarization when its metabolic elimination was impaired, such as by liver disease or drugs that inhibit the 3A family of cytochrome P450. In vitro studies indicate that this action is due to blockade of one or more of the cardiac potassium channels that determine the duration of the action potential. Prescription guidelines are now available to reduce the risk of developing arrhythmias with these two drugs. Two agents recently marketed in the United States, Ioratidine and cetirizine, appear to lack the ability to prolong repolarization and induce torsades de pointes. Evaluation of the potential cardiac actions of investigational antihistamines is essential and may be of value for some of the older conventional agents. PMID- 8725390 TI - New pharmacological strategies for pain relief. AB - The recent advances made in elucidating the processes of nociception have altered the way that chronic pain therapy and analgesic drug development are approached. Recent studies have highlighted new targets for drug discovery, including inhibition of inflammatory mediators (kinins, growth factors), newly expressed proteins (B1 receptors, COX-2), and blockers of afferent fiber activity (capsaicin analogues, ion channel blockers). In the CNS, a further multiplicity of strategies can be pursued, including the development of antagonists of specific neuropeptide and glutamate receptors or agonists for purine and amine receptors. Such drugs will inevitably supplement or replace conventional NSAID and opioid analgesics. Further characterization of gene regulation will allow the development of drugs that genetically modify cellular activity altered by chronic pain conditions. PMID- 8725391 TI - Angiotensin receptors and their therapeutic implications. AB - Angiotensin II is a multifunctional hormone that exerts its effects by interacting will cell surface receptors. Two major subtypes of receptors (AT1 and AT2) have been distinguished by pharmacological and molecular biological techniques. AT1 receptors have been further subdivided into AT1A and AT1B receptors. Several other isoforms have been found, notably in nonmammalian systems, but further information is necessary before definitive classification can be made. AT1 receptors mediate most known functions of angiotensin II, while AT2 receptors may be important developmentally. The molecular, structural, and biochemical characteristics of these receptors have been described, as well as the factors that regulate their expression. This receptor system has been implicated in several cardiovascular diseases, including hypertension, restenosis after angioplasty, cardiac hypertrophy, heart failure, myocardial infarction, and ventricular remodeling. Structural analysis of AT receptors may provide the basis for the development of new therapeutic agents with enhanced specificity for the treatment of these diseases. PMID- 8725392 TI - Assessment of follicle destruction in chemical-induced ovarian toxicity. AB - The ovary plays a pivotal role in the regulation of reproductive function in females. Development, maturation, and ovulation of oocytes occur within ovarian follicles. Females are born with a finite number of undeveloped, primordial follicles. Chemicals that destroy oocytes contained in these follicles can produce premature ovarian failure (menopause) because once a primordial follicle is destroyed, it cannot be replaced. This article focuses on chemicals that have been shown to destroy these small follicles in laboratory animals. An ovotoxic compound of particular interest is 4-vinylcyclohexene (VCH) and its epoxide metabolites. The specific information discussed here relates to various aspects of VCH-inuced ovarian toxicity. This includes species specificity, disposition and metabolism, direct follicular effects, bioactivity of metabolites, and long term responses to exposure. Using these studies as a model approach, a method for evaluation of the ovotoxic potential of other chemicals can be designed. PMID- 8725393 TI - CNS effects and abuse liability of anabolic-androgenic steroids. AB - Anabolic-androgenic steroids (AAS) have become more widely used and abused not only by elite athletes, but by nonathletes as well. It appears that these drugs are being used for cosmetic purposes as well as for promoting aggressive behavior. However, AASs may also be used in combination with other drugs such as ethyl alcohol and cocaine--a practice that may contribute to the observed increase in aggressive behavior. As there are no reliable animal models of AAS self-administration, it has been difficult to characterize the abuse liability and physical dependence potential of this class of drugs. A better understanding of neuroactive steroids, improved methods for assessing AAS abuse, and a revised interpretation of drug-seeking behavior is needed to develop improved treatment strategies for this emerging health-related problem. PMID- 8725394 TI - Pathophysiological basis of vulnerability to drug abuse: role of an interaction between stress, glucocorticoids, and dopaminergic neurons. AB - Research on drug abuse has recently focused on understanding the vulnerability to develop addiction that is present in certain individuals. These investigations suggest that addiction results from an interaction between drugs and specific individual substrates. Differences in the propensity to develop drug intake can be demonstrated in animals with equal access to drugs under stable laboratory conditions and can be predicted by drug-independent behaviors. Stress, corticosterone, and mesencephalic dopaminergic neurons seem to be organized in a pathophysiological chain determining such a vulnerability. An increased corticosterone secretion, or a higher sensitivity to the effects of this hormone, either naturally present in certain individuals or induced by stress in others, increases the vulnerability to develop drug intake, via an enhancement of the activity of mesencephalic dopaminergic neurons. These findings suggest that addiction therapies should counteract the biological peculiarity that leads some individuals to respond in a pathophysiological way to drugs. PMID- 8725395 TI - Opioid receptor types and subtypes: the delta receptor as a model. AB - Since the discovery of opioid receptors over two decades ago, an increasing body of work has emerged supporting the concept of multiple opioid receptors. Molecular cloning has identified three opioid receptor types--mu, delta, and kappa--confirming pharmacological studies that previously postulated the existence of these three receptors. The cloned opioid receptors are highly homologous and belong to the family of seven-transmembrane, G protein-coupled receptors. With the development of novel opioid ligands, subtypes of the mu, delta, and kappa receptors have been proposed, although the molecular basis of these subtypes has not been elucidated. In this review, we present the pharmacological data supporting the concept of multiple delta opioid receptor subtypes and offer hypothetical mechanisms which might generate these "subtypes." PMID- 8725396 TI - Cyclic nucleotide phosphodiesterases and vascular smooth muscle. AB - At least 30 different phosphodiesterase (PDE) enzymes have now been identified in mammalian tissues and cells, many of which are products of separate genes. These different isoenzyme forms can be subdivided into seven families based on their genetic and functional characteristics. Relatively specific inhibitors are available for at least five of these PDE families. A functional classification based on substrate specificity, regulatory properties, and sensitivity to inhibition by isozyme- and tissue-selective inhibitors can be used in describing the PDEs of vascular smooth muscle. Inhibition of these PDEs, especially with inhibitors of the PDE3 isoform, promotes vascular relaxation, particularly if the preparation of smooth muscle has been preconracted. For the most part, the drugs appear to act directly on smooth muscle; their effects are usually observed in endothelium-denuded preparations. In addition to their cardiotonic properties, many PDE3 inhibitors possess antiplatelet and thrombolytic activities, thereby suggesting the potential benefit of these drugs in treating diseases of the cardiovascular system. Isozyme- and cell-specific drugs have been shown to alter the synthetic state (i.e. proliferative phenotype) of smooth muscle cultures toward the appearance of the contractile phenotype. This suggests the possible use of selective PDE inhibitors to minimize the problem of restenosis seen after angioplasty. The development of novel methods to deliver more potent and selective PDE inhibitors to individual cell types and subcellular locales will lead to new therapeutic uses for this class of drugs in diseases of the cardiovascular system. PMID- 8725397 TI - Diversity and selectivity of receptor-G protein interaction. AB - Cells in a living organism communicate with each other through extracellular molecules such as hormones, neurotransmitters, and growth factors. The majority of these molecules transmit their signal by interacting with a three-protein transmembrane signal transduction system whose single components interact sequentially and reversibly. Agonist binding to a heptahelical receptor results in activation of heterotrimeric guanine nucleotide-binding proteins (G proteins) that modulate the activity of one or more effector systems. Considering that hundreds of G protein-coupled receptors transduce signals by interacting with a limited repertoire of G proteins, the question of specificity governing the coupling of receptors to G proteins arises. The conceptualization of signal transduction pathways in a linear fashion (one receptor coupling to one G protein that activates one effector) is inadequate to explain experimental results. In the present review, G protein-mediated signal transduction is depicted as a complex signaling network with divergent and convergent pathways at each transduction level, i.e. receptor, G protein, and effector. The recent realization that "classical" signaling pathways appear to be activated in parallel with signaling cascades primarily described for growth factors and cytokines adds an additional level of intriguing complexity. PMID- 8725398 TI - Complexity and diversity of mammalian adenylyl cyclases. AB - Molecular cloning has permitted identification of several novel isoforms of mammalian adenylyl cyclase; these proteins now comprise a family of at least 10. All of the membrane-bound enzymes are activated by the alpha subunit of G alpha, a receptor-regulated, heterotrimeric guanine nucleotide-binding protein, and by the diterpene forskolin. Certain cyclases are also activated by Ca(2+) calmodulin, while some are inhibited by the alpha subunits of the three Gi proteins. The discovery of new isoforms has also revealed unanticipated mechanisms of regulation, including activation or inhibition by the G-protein beta gamma subunit complex, inhibition by G(o) alpha, inhibition by Ca2+, and phosphorylation by protein kinases C and A. The effects of activators are often highly synergistic or conditional, suggesting function of these enyzmes as coincidence detectors. The plethora of receptors, G proteins, and adenylyl cyclases permits assembly of very complex signaling systems with a wide variety of integrative characteristics. PMID- 8725399 TI - Regulation of phosphoinositide phospholipases by hormones, neurotransmitters, and other agonists linked to G proteins. AB - The actions of many hormones, neurotransmitters, and growth factors are mediated by the hydrolysis of phosphatidylinositol 4,5-bisphosphate catalyzed by specific isozymes of phospholipase C. This hydrolysis releases inositol 1,4,5 trisphosphate, which mobilizes Ca2+ ions from components of the endoplasmic reticulum, and 1,2-diacylglycerol, which activates isozymes of protein kinase C. The hormones and neurotransmitters activate beta-isozymes of phospholipase C through receptors that have seven transmembrane segments and couple to G proteins of the Gq and Gi/o families. Activation of phospholipase C by the Gq family involves their alpha-subunits, whereas activation by the Gi/o family involves their beta gamma-subunits. The growth factors activate gamma-isozymes of phospholipase C through receptors that become autophosphorylated due to their stimulated tyrosine kinase activity and provide binding sites for the Src homology domains of the isozymes. The molecular mechanisms by which agonists activate phopholipase isozymes are described in detail. PMID- 8725400 TI - Imidazoline receptors and their endogenous ligands. AB - Imidazoline (I) receptors constitute a family of nonadrenergic high-affinity binding sites for clonidine, idazoxan, and allied drugs. One major subclass, the I1 receptors, whose subcellular distribution and signal transduction mechanisms are uncertain, partly mediates the central hypotensive actions of clonidine-like drugs. The I2 receptors, another subclass, are mitochondrial, not G protein coupled, and have diversified functions. Several endogenous ligands for I receptors, collectively termed clonidine-displacing substances (CDSs), have been detected in tissues and serum, but the structure of only one, agmatine (decarboxylated arginine), is known. Agmatine, widely distributed and bioactive, binds, like clonidine, to alpha 2-adrenergic and I receptors of all subclasses. The presence of agmatine and its biosynthetic enzyme in synaptosomes and specific neuronal pathways as well as serum suggests that it may be a novel neurotransmitter/hormone. Another CDS that binds to I receptors and to antibodies to imidazoline drugs has been detected, but its structure is unknown. PMID- 8725401 TI - Structural mechanisms of HIV drug resistance. AB - Antiviral therapy for AIDS has focused on the discovery and design of inhibitors for two main enzyme targets of the human immunodeficiency virus type 1 (HIV)- reverse transcriptase (RT) and protease (PR). Despite several classes of promising new anti-HIV agents, the clinical emergence of drug-resistant variants of HIV has severely limited the long-term effectiveness of these drugs. Genetic analysis of resistant virus has identified a number of critical mutations in the RT and PR genes. Structural analysis of inhibitor-enzyme complexes and mutational modeling studies are leading to a better understanding of how these drug resistance mutations exert their effects at a structural level. These insights have implications of the design of new drugs and therapeutic strategies to combat drug resistance to AIDS. PMID- 8725402 TI - Breast cancer and environmental risk factors: epidemiological and experimental findings. AB - Breast cancer has long been associated with reproductive hormone exposures. Recently, greater attention has been focused on environmental exposures that may be responsible for some proportion of breast cancer incidence. Several etiologic aspects are discussed. A number of chemicals induce breast cancer in rodents- including solvents, pesticides, and polycyclic aromatic hydrocarbons--and these might serve as leads for studies in humans. In women, strong links have been established between breast cancer risk and ionizing radiation. Evidence for nonionizing radiation (electromagnetic field) exposures and breast cancer is suggestive, albeit limited. Occupational exposures have not been identified as breast cancer risks, but several associations need further study, including solvents and pesticides. Time of life when exposures take place is important, and this claim is strongly supported by data on cigarette smoking and radiation. Also, basic research has demonstrated that mammary tissue is more susceptible to carcinogenesis at certain periods of breast development. Likewise, prenatal, neonatal, and adolescent exposures deserve continuing attention. Research on etiology of breast cancer should measure environmental exposures and take into account the time of life at which these occur. Complex interactions between exogenous and endogenous carcinogenic agents need further focus, as modulated by varying genetically determined individual susceptibilities. PMID- 8725403 TI - Pharmacology of nicotine: addiction and therapeutics. AB - Nicotine maintains tobacco addiction and has therapeutic utility to aid smoking cessation and possibly to treat other medical diseases. Nicotine acts on nicotinic cholinergic receptors, which demonstrate diversity in subunit structure, function, and distribution within the nervous system, presumably mediating the complex actions of nicotine described in tobacco users. The effects of nicotine in people are influenced by the rate and route of dosing and by the development of tolerance. The metabolism of nicotine is now well characterized in humans. A few individuals with deficient C-oxidation of nicotine, unusually slow metabolism of nicotine, and little generation of cotinine have been described. Nicotine affects most organ systems in the body, although its contribution to smoking-related disease is still unclear. Nicotine as a medication is currently available as a gum, a transdermal delivery device, and a nasal spray, all of which are used for smoking cessation. Nicotine is also being investigated for therapy of ulcerative colitis, Alzheimer's disease, Parkinson's disease, Tourette's syndrome, sleep apnea, and attention deficit disorder. PMID- 8725404 TI - Insulin signal transduction and the IRS proteins. AB - Insulin controls organismal and cellular physiology by initiating numerous intracellular signals. Insulin first binds the extracellular domain of the insulin receptor, which activates the receptor's intracellular tyrosine kinase. Receptor-mediated phosphorylation of the IRS proteins is required for the propagation of signals for mitogenesis, glucose transport, and numerous other biological and biochemical events during insulin signaling. IRS proteins also mediate signaling by a subset of other growth factor and cytokine receptors; recognition and phosphorylation by specific receptors appears to be mediated by the PH and PTB domains of the IRS proteins. The best understood mechanism of IRS protein-mediated signaling is the binding of SH2 domain-containing signaling molecules (such as PI 3'-kinase) by tyrosine phosphorylation sites on IRS proteins. Other paradigms of IRS-protein signaling are beginning to emerge, however, and these exciting molecules promise to teach us much in the next few years. PMID- 8725405 TI - Exocytosis: proteins and perturbations. AB - Exocytosis is the primary means of cellular secretion. Because exocytosis involves fusion between the plasma membrane and the membrane of secretory vesicles, it is likely that proteins on these two membranes, as well as additional proteins in cellular cytoplasm, mediate exocytosis. Although we know much about the proteins of secretory cells, we still have much to learn about how these proteins participate in exocytosis; in no case has an unambiguous exocytotic function been assigned to any of these proteins. To identify the roles of proteins in exocytosis it is necessary to perturb protein function in living secretory cells. We review a number of perturbation strategies and summarize what this approach has taught us about the functional roles of proteins in exocytosis, concluding with a molecular model of protein dynamics during exocytosis. PMID- 8725406 TI - The Bin1 gene localizes to human chromosome 2q14 by PCR analysis of somatic cell hybrids and fluorescence in situ hybridization. PMID- 8725407 TI - Training, certification, and credentialing in transesophageal echocardiography. PMID- 8725408 TI - Validation of a test of competence in transesophageal echocardiography. AB - OBJECTIVE: A test was developed that is based on intraoperative findings to evaluate knowledge of intraoperative echocardiography. This study examines the performance of attending anesthesiologists and residents at various levels of training in order to validate the test's ability to measure physician competence in intraoperative echocardiographic diagnosis. DESIGN: This study was a prospective evaluation of a test of competence in transesophageal echocardiography. SETTING: Two university medical centers. PARTICIPANTS: Volunteer anesthesia faculty and residents. INTERVENTIONS: The participants took an echocardiographic examination that consisted of 34 "K"-type multiple-choice questions and a 45-second video-loop sequence for each question. The video sequences were chosen for their high quality and unambiguous representation of both normal and pathologic images obtained by transverse transesophageal echocardiographic imaging. The questions were written by experienced echocardiographers. The test was administered to 25 individuals at two academic institutions: 11 residents with minimal transesophageal echocardiography exposure and 14 faculty who were relatively experienced with transesophageal echocardiography. All of the residents repeated the examination at the end of their third clinical anesthesia year, which included transesophageal echocardiography training. The differences between the groups' scores were analyzed using the Kruskal-Wallis test and Wilcoxon's rank-sum test. To correct for the multiple comparisons, p < 0.025 was deemed significant. MEASUREMENTS AND MAIN RESULTS. Before their transesophageal echocardiography training, the residents scored significantly lower than the faculty (p < 0.002). After 1 year of training, their scores significantly increased (p = 0.021), and their scores were not significantly different from the faculty level (p = 0.052). CONCLUSIONS: Test performance differed according to level of experience. This suggests that the test is a valid measure of intraoperative transesophageal echocardiography competence. PMID- 8725409 TI - Detection of aortic emboli by transesophageal echocardiography during coronary artery bypass surgery. AB - OBJECTIVE: The purpose of this study was to determine whether emboli can be detected within the aortic lumen in patients undergoing coronary artery bypass surgery (CABG) and to relate the appearance of emboli to specific operative events. DESIGN: Twenty patients were prospectively studied intra-operatively. SETTING: Subjects were inpatients in an academic medical center. PARTICIPANTS: All participants were scheduled for elective, isolated CABG. INTERVENTIONS: Patients were continuously monitored using transesophageal echocardiography (TEE) from aortic cannulation to bypass discontinuation. After completion of the aortic examination, the probe was focused at the level of the aortic arch, just before the takeoff of the left subclavian artery. Emboli were defined as echogenic intraluminal signals not present in the same position on consecutive cross sectional frames. RESULTS: Intraluminal emboli were detected in all subjects, with a mean number of 535 and range of 8 to 1,885. Embolization was unevenly distributed through the procedure. A mean of 224 (42%) of 535 were detected within 4 minutes of aortic cross-clamp release and another 140 (24%) appeared after partial occlusion clamp release. Together, clamp placement and release represented 84% of all emboli. Emboli detected after clamp release were large, echodense particles easily distinguishable from the small, indistinct, poorly echogenic signals observed at bypass initiation. CONCLUSIONS: Emboli can be visualized within the aortic lumen during CABG. Confirming previous reports, the majority of emboli detected are related to manipulation of aortic clamps. The composition and clinical significance of embolic material are unclear. The value of intraoperative TEE monitoring in predicting neurologic outcome remains to be determined. PMID- 8725410 TI - Hepatic blood flow and right ventricular function during cardiac surgery assessed by transesophageal echocardiography. AB - OBJECTIVE: To determine the effect of cardiopulmonary bypass (CPB) on hepatic blood flow (HBF) and the hepatic venous flow pattern. DESIGN: Single-arm prospective study. SETTING: University hospital operating room and intensive care unit. PARTICIPANTS: Eight patients ranging in age from 57 to 73 years undergoing cardiac surgery. INTERVENTIONS: Transesophageal echocardiography (TEE) was used to assess HBF before, during, and after CPB by pulsed-wave Doppler ultrasound recordings of hepatic venous flow velocity and two-dimensional recordings of the hepatic vein diameter. Hepatic vein oxygenation was monitored by hepatic vein catheterization, and gastric intramucosal pH (pHi) was followed by tonometry. MEASUREMENTS AND MAIN RESULTS: The HBF was unchanged after the start of CPB but was reduced from the baseline value 415 (standard error of the mean 40) mL/min to 225 (25) mL/min during hypothermic CPB (p < 0.05). Cardiac index, right ventricular ejection fraction, and arterial and tonometric pH were essentially unchanged during the study period. Hepatic vein and mixed venous saturation were unchanged compared to control during CPB and were reduced at 2 and 3 hours after CPB (p < 0.01). Six of the patients had a normal predominant systolic flow pattern before surgery. In the postoperative period, seven patients showed an abnormal predominant diastolic filling pattern. CONCLUSIONS: TEE represents a useful tool in assessing changes in the hepatic blood flow. The HBF was reduced during hypothermic CPB, but this was not accompanied by a reduced pHi. The changes in the venous flow pattern with a reduction in systolic flow could be explained by impaired atrial relaxation. PMID- 8725411 TI - The agreement between ventricular volumes and ejection fraction by transesophageal echocardiography or a combined radionuclear and thermodilution technique in patients after coronary artery surgery. AB - OBJECTIVES: To study the reproducibility of and agreement between perioperative transesophageal echocardiographic (TEE) and radionuclide (RN) assessment of ventricular volumes and ejection fraction (EF). DESIGN: A prospective, blinded comparison of two methods of measurement. SETTING: A surgical intensive care unit in a university hospital. PARTICIPANTS: Patients after coronary artery bypass surgery. INTERVENTIONS: Left ventricular volumes and ejection fraction were simultaneously measured by radionuclide ventriculography and transesophageal echocardiography. Two sets of measurements were made in rapid succession with a third set after an interval. RESULTS: Reproducibilities of EF and ventricular volumes by TEE and RN were similar. At each set of measurements, the bias for radionuclide EF and TEE Simpson's rule EF, 0.03 +/- 0.05, 0 +/- 0.06, -0.01 +/- 0.07, respectively, for radionuclide EF and TEE area length EF 0.01 +/- 0.05, 0.01 +/- 0.05, -0.03 +/- 0.08, respectively, were significantly less than for radionuclide EF and TEE FAC 0.07 +/- 0.05, 0.05 +/- 0.05, 0.03 +/- 0.09. Poor agreement was observed between RNTD-EDV, and both of the TEE EDV measurements. CONCLUSION: EF measured by TEE area length and Simpson's rule method are as reproducible as TEE FAC and are more accurate estimates of RN EF. Poor agreement between methods of measuring end-diastolic volume was observed. PMID- 8725412 TI - Cardiac complications in noncardiac surgery: value of dobutamine stress echocardiography versus dipyridamole thallium imaging. AB - OBJECTIVES: The objective of this study was to determine the relative value of dobutamine stress echocardiography (DSE) and dipyridamole thallium imaging (DT) in the preoperative assessment of cardiovascular risk before noncardiac surgery. DESIGN: Prospectively DSE was performed in patients who had undergone DT as a part of their preoperative evaluation. SETTING: A large urban veterans' affairs medical center. PARTICIPANTS: Thirty-seven patients undergoing major noncardiac surgery were assessed for complications during a 1-month follow-up period. INTERVENTIONS: Both DSE and DT were performed before surgery. The medium interval between the two tests were 15.5 days. MEASUREMENTS: Left ventricular wall motion was assessed at baseline and peak dobutamine dose in a standard fashion. Wall motion was scored and indexed using a 16-segment model. A positive DSE was defined as failure of augmentation, new or worsening of baseline wall motion abnormalities in two or more contiguous segments. Myocardial perfusion studies after DT were performed according to conventional method. A positive DT was defined as a reversible perfusion defect, increased lung uptake, and/or transient left ventricular dilatation. Complications were defined as myocardial infarction or cardiac death occurring as a result of the operation, or need for revascularization before surgery. RESULTS: DSE was positive in 19 patients, whereas DT was positive in 25 patients. Fourteen patients had both an abnormal DSE and DT. Five patients had major postoperative cardiac complications: fetal myocardial infarction (1); fatal cardiac arrest (1); and severe coronary artery disease necessitating coronary artery bypass surgery (2) or percutaneous transluminal coronary angioplasty (1). DSE was positive in all 5 (100%), whereas DT was positive in 4 of 5 (80%) patients with complications. The sensitivity for each test was comparable: for DSE it was 100% (95% C.I. 56% to 100%) and for DT 80% (37% to 96%). Specificity for DSE (60%, 43%-74%) was somewhat higher than DT (38%, 24% to 54%), although this did not reach statistical significance (p = 0.06). CONCLUSIONS: The ability of DSE to predict major cardiac complications related to noncardiac surgery appears to be similar to DT and may be used as an alternative to DT imaging in the preoperative risk assessment of patients undergoing noncardiac surgery. PMID- 8725413 TI - Afterdrop after hypothermic cardiopulmonary bypass: the value of tympanic membrane temperature monitoring. AB - OBJECTIVES: After weaning from cardiopulmonary bypass (CPB), a decrease in nasopharyngeal temperature (NPT) occurs (afterdrop). The pathophysiology of the afterdrop remains unclear: It might be caused by either inadequate total body rewarming on CPB or to heterogenous distribution of heat during CPB, with subsequent redistribution of heat from the warmer core to the cooler shell tissues. The study objectives were (1) to determine whether post-CPB afterdrop is the result of a negative CPB thermal balance, and (2) to investigate which sites (if any) could best predict the afterdrop. DESIGN: Prospective evaluation using within-patient comparisons during CPB cooling, CPB rewarming, and 45 minutes post CPB. SETTING: Adult patients gave informed consent before a cardiac surgical procedure in a university hospital. PARTICIPANTS: Eight patients undergoing CABG or valvular replacement with hypothermic CPB (NPT near 29 degrees C) and standardized general anesthesia. INTERVENTIONS: Each patient was studied with temperature monitors (Mon-a-therm 7000; Mallinckrodt-Medexel, Gemenos, France) attached to disposable thermocouple probes placed as follows: urinary bladder, rectum, deltoid, esophagus, nasopharynx, tympanic membrane, and four skin sites. In addition, the temperatures from the thermistors of the pulmonary artery catheter, and the arterial and venous lines of the CPB circuit were considered. Thirteen sites for monitoring temperature were studied. MEASUREMENTS AND MAIN RESULTS: Temperatures were recorded every 5 minutes, from the beginning of CPB to the 45th minute after CPB, and thermal exchanges were calculated: change in body heat (QBH), thermal exchanges between the patient and the pump (QCPB), metabolic heat production (Qm) (equal to calculated VO2 at the pump level), and heat loss to the environment (QS) (equal to QBH-QCPB-Qm). Thermal exchanges were obtained in six patients during the plateaus of cooling and rewarming, during the whole CPB phase, and after CPB. It was found that despite a change in QBH during rewarming (1,017 +/- 88 kJ) that was slightly greater than during cooling (-1,008 +/- 104 kJ) (mean +/- SEM), a significant decrease in post-CPB "core" temperature occurred (afterdrop: -1.4 degrees C). Magnitude of the afterdrop was directly related to the magnitude of tympanic membrane cooling and was negatively correlated to the temperature difference between the warmest site (tympanic membrane) and the coolest site (cutaneous thigh temperature) observed at the end of rewarming (r = -0.667; p < 0.05). CONCLUSIONS: It is suggested that besides post-CPB heat loss, redistribution of heat may be involved in the mechanism of the afterdrop and that measurements of tympanic membrane and cutaneous thigh temperatures are the best monitors of adequacy of rewarming during CPB. PMID- 8725414 TI - Hypothermic versus normothermic cardiopulmonary bypass: influence on circulating adhesion molecules. AB - OBJECTIVE: Cardiopulmonary bypass (CPB) may result in a whole-body inflammatory response with the risk of subsequent development of organ failure. Leukocyte endothelial binding followed by neutrophil migration appear to play a central role. This process is markedly influenced by adhesion molecules. Whether plasma levels of circulating adhesion molecules are beneficially influenced by hypothermic CPB was studied in patients undergoing either hypothermic or normothermic CPB. DESIGN: Prospective, randomized study. SETTING: Single Institutional, clinical investigation in a cardiac anesthesia department of a university hospital. PARTICIPANTS: 30 patients scheduled for elective aortocoronary artery bypass grafting. INTERVENTIONS: The patients were prospectively and randomly divided into two groups: group 1 underwent hypothermic CPB (rectal temperature 27 to 28 degrees C; n = 15) and group 2 normothermic CPB (rectal temperature > 36 degrees C; n = 15). MEASUREMENTS AND MAIN RESULTS: Plasma levels of circulating (soluble) adhesion molecules (endothelial leukocyte adhesion molecules [sELAM-1], vascular cell adhesion molecule-1 [sVCAM-1], intercellular adhesion molecule-1 [sICAM-1], and granule membrane protein 140 [sGMP-140]) were measured from arterial blood samples using enzyme-linked immunosorbent assays (ELISA) after induction of anesthesia (= baseline values), after weaning from bypass, at the end of surgery, 5 hours after the end of CPB, and on the morning of the first postoperative day. Mean rectal temperature of group 1 was 27.2 +/- 0.4 degrees C and 36.7 +/- 0.4 degrees C in group 2. In both groups, plasma levels of sELAM-1 were significantly higher than baseline only 5 hours after CPB. sICAM-1 increased until the first postoperative day (group 1: +35%; group 2: +37%) without, however, exceeding the normal range. sVCAM-1 plasma levels increased after CPB (group 1: +56%; group 2: +40%). At the end of surgery and 5 hours after CPB, sGMP-140 plasma levels were significantly higher in the hypothermic (increase from 301 +/- 34 to 582 +/- 57 ng/mL) than in the normothermic patients (increase from 310 +/- 45 to 480 +/- 32 ng/mL). On the first postoperative day, both groups showed similar, significantly elevated plasma levels of sGMP-140. CONCLUSIONS: Plasma levels of circulating adhesion molecules sELAM-1, sICAM-1, and sVCAM-1 did not differ between hypothermic and normothermic CPB, indicating no differences in endothelial activation between the two groups. Only sGMP-140 plasma levels were increased more after hypothermic CPB. Additional influences of hypothermia on the coagulation system might have contributed to the higher sGMP-140 plasma levels of these patients. The definite role of circulating adhesion molecules in cardiac surgery patients remains to be elucidated. PMID- 8725415 TI - Comparison of hetastarch with albumin for postoperative volume expansion in children after cardiopulmonary bypass. AB - OBJECTIVE: Hetastarch has been studied as a volume expander in adults after cardiopulmonary bypass (CPB) and in recommended dosages has not altered coagulation studies or increased clinical bleeding. Hetastarch was compared with albumin in children after CPB to determine whether hetastarch use was associated with increased clinical bleeding or alteration of coagulation studies. DESIGN: Randomized double-blinded study. SETTING: University-affiliated children's hospital. PARTICIPANTS: Forty-seven children age 1 year or greater (mean 72.8 months; range 12 months to 15.5 years) scheduled for repair of congenital heart disease with moderate hypothermia were randomized to receive hetastarch or albumin as a postoperative volume expander during the first 24 hours after surgery. INTERVENTIONS: Thirty-eight children required colloid replacement therapy. Blood pressure, central venous pressure, urine output, and chest tube drainage were used to determine colloid requirement. MEASUREMENTS AND MAIN RESULTS: Clinical bleeding and laboratory studies of coagulation were evaluated as were requirements for colloid, crystalloid, and blood products. Twenty children received 6% hetastarch, and 18 received 5% albumin. No differences were found in the amount of replacement fluids required, or in coagulation parameters in children receiving 20 mL/kg or less of either colloid replacement therapy. An increase in prothrombin time was demonstrated in children who received greater than 20 mL/kg of 6% hetastarch (p = 0.006); however, no difference in clinical bleeding or blood product requirement was demonstrated between the hetastarch or albumin groups receiving more than 20 mL/kg. CONCLUSION: This study demonstrated that 6% hetastarch is safe and an effective plasma volume expander in the postoperative management of children, using volumes up to 20 mL/kg. Close laboratory monitoring and careful evaluation of clinical bleeding are suggested when larger doses of hetastarch are administered because of prolongation of the prothrombin time with more than 20 mL/kg of hetastarch. PMID- 8725416 TI - In vitro uptake and elimination of isoflurane by different membrane oxygenators. AB - OBJECTIVE: This study was designed to investigate the effect of membrane oxygenator design and composition on the uptake and elimination of isoflurane. DESIGN: Prospective, in vitro laboratory study. SETTING: Bioengineering laboratory. PARTICIPANTS: Three types of membrane oxygenator were tested: the SM 35 (polydimethylsiloxane in sheet form), the CML (polypropylene in sheet form), and the SAFE II (polypropylene in hollow-fiber form). The oxygenators were incorporated into a standard cardiopulmonary bypass circuit. INTERVENTIONS: Isoflurane was added to the oxygenator input gas and measured in exhaust gas and in (bovine) blood leaving the oxygenator at 1, 2, 3, 5, 7, 10, 15, and 20 minutes. The isoflurane vaporizer was then turned off, and samples were obtained at the same time intervals. The experiment was performed at 28 degrees C and 37 degrees C. MEASUREMENTS AND MAIN RESULTS: Uptake and elimination of isoflurane were slower via the SM-35 compared with the CML and the SAFE II (p < 0.01). CONCLUSIONS: If isoflurane is administered during cardiopulmonary bypass, knowledge of the influence of oxygenator membrane composition on its pharmacokinetics is essential if patient awareness and unexpected cardiovascular depression are to be avoided. PMID- 8725417 TI - The effect of midazolam at two plasma concentrations of hemodynamics and sufentanil requirement in coronary artery surgery. AB - OBJECTIVES: In this study, the hemodynamics and sufentanil requirement were compared at two midazolam target plasma concentrations in patients undergoing coronary artery bypass grafting (CABG). DESIGN: Prospective, randomized study. SETTING: University hospital, single institution. PARTICIPANTS: Patients undergoing CABG. INTERVENTIONS: Patients were randomly assigned to receive midazolam at a target plasma concentration of 150 ng/mL (group 1; n = 10) or 300 ng/mL (group 2; n = 10). Sufentanil infusion was titrated to maintain hemodynamic stability, defined as mean arterial pressure within 15% of baseline values. All patients received preoperative beta-blocking agents. Arterial blood samples of midazolam and sufentanil were analyzed by high-performance liquid chromatography and radioimmunoassay, respectively. MEASUREMENTS AND MAIN RESULTS: The mean dose of sufentanil (7.5 +/- 1.7 microgram/kg in group 1 v 7.2 +/- 2.5 micrograms/kg in group 2) did not differ. There were no significant differences in hemodynamics between the groups in the period before or after cardiopulmonary bypass (CPB). Before CPB, in two patients in each group, hypertension was controlled with sufentanil only. One patient in group 1 required a vasodilator in addition to sufentanil. No ischemic events occurred before CPB. After CPB, one patient in group 2 required a vasodilator to control hypertension. Two patients in group 2 required treatment with nitroglycerin for myocardial ischemia. Stable plasma concentrations of sufentanil and midazolam were obtained during and after CPB. The midazolam infusion was continued in both groups at a rate of 1.25 micrograms/kg/min during the first 4 postoperative hours. The time to awakening did not differ between the groups (100 +/- 58 minutes in group 1 v 173 +/- 147 minutes in group 2) nor did the plasma concentrations of midazolam (96 +/- 28 ng/mL v 108 +/- 42 ng/mL) at the time of awakening. Intraoperative awareness was not reported. CONCLUSION: In patients undergoing CABG, good hemodynamic control with a similar incidence of hemodynamic interventions was observed at midazolam target plasma concentrations of 150 and 300 ng/mL when coadministered with sufentanil. The sufentanil requirement was identical in both groups. This study suggests that a midazolam plasma concentration of 150 ng/mL is sufficient to provide satisfactory hemodynamic control and to avoid intraoperative awareness. PMID- 8725418 TI - Dual vasoactive effects of tolazoline on rabbit pulmonary arteries. AB - OBJECTIVES: To determine the vasoactive effects of tolazoline on isolated rabbit pulmonary arteries. DESIGN: Prospective, in vitro, randomized, controlled trial. SETTING: Experimental laboratory in a university-affiliated hospital. PARTICIPANTS: New Zealand White Rabbits. INTERVENTIONS: The pulmonary artery rings were obtained via thoracotomy. Their vasoactive responses were assessed in the presence and absence of intact endothelium and with or without precontraction by norepinephrine (NE, 3 x 10(-6) M) or potassium chloride (KCl, 3 x 10(-2) M). Using a tissue bath preparation, cumulative concentration response curves of tolazoline were obtained at different concentrations (10(-9) to 10(-4) M) after a period of stabilization. MEASUREMENTS AND MAIN RESULTS: Tolazoline caused vasoconstriction of isolated pulmonary arteries without any pretreatment. The magnitude of the constriction was dose related and reached 300 g/g wet tissue at a concentration of 10(-4) M. On KCl-precontracted pulmonary arteries, tolazoline caused significant dose-related vasoconstriction. On the NE-precontracted vessel rings, it elicited significant dose-dependent vasodilation up to 60% relaxation at 10(-5) M. All the above effects were endothelium independent. CONCLUSIONS: Tolazoline has dual endothelium-independent vasoactive effects, causing vasoconstriction on isolated rabbit pulmonary arteries, either untreated or precontracted with KCl, and vasodilation on those precontracted with NE. Tolazoline may act as a competitive alpha-adrenoceptor blocking agent. PMID- 8725419 TI - An alternative to radioactive microspheres for measuring regional myocardial blood flow, Part 1: Colored microspheres. AB - OBJECTIVE: To compare measurements of regional myocardial blood flow between color and radioactive microspheres. DESIGN: Prospective, randomized, controlled. SETTING: University research laboratory. PARTICIPANTS: Pigs. INTERVENTIONS: Pigs underwent constriction of the left anterior descending artery, either incremental and then 0 constriction with epinephrine, 0.5 to 3 mu/kg/min (n = 5; "variable") or only 0% and 100% constriction without epinephrine (n = 4, "single"). Radioactive and color microspheres were injected simultaneously. For variable constriction, 5 colors (3 x 10(6)/mL) were tested in random order and, for single, red and yellow (6 x 10(6)/2 mL). MEASUREMENTS AND MAIN RESULTS: Measurements of regional endocardial, epicardial, and transmural myocardial blood flow were compared by regression analysis (linear and nonlinear). With radioactive measurements as the point of reference, when regional flow was 50 to 150 mL/min/100 g, correlation was high (r = 0.85), although regression slope was low. With endocardial and epicardial flow between 30 and 100 mL/min/100 g, correlation was close (r = 0.84). Overall nonlinear correlation was higher with single than variable constriction (r = 0.72). When regional flow was less than 100 mL/min/100 g, linear correlation was r = 0.72. When transmural flow measured by color microspheres was less than 25 mL/min/100 g, correlation was high (r = 0.86) but, with endocardial or epicardial flow, low (r = 0.67). When transmural flow was greater than 100 mL/min/100 g, correlation was extremely low (r = 0.1; n = 26 data points). The overall correlations for regional endocardial and epicardial flows were also low, except in the ischemic zone. CONCLUSIONS: Color and radioactive measurements correlate well during moderate and ischemic regional myocardial blood flow, ischemic blood flow requiring a higher concentration of color microspheres. A major limitation of using color microspheres is imprecision when flow is greater than 150 mL/min 100 g. PMID- 8725420 TI - An alternative to radioactive microsphere for measuring regional myocardial blood flow, Part 2: Laser-Doppler perfusion monitor. AB - OBJECTIVE: To compare measurements of regional myocardial blood flow volume between microsphere measurement of regional flow (0.5- to 2- g tissue sampling) and a potential alternative measure, local flow (1 mm3) in the microcirculation measured by laser-Doppler perfusion monitor. DESIGN: Prospective, randomized, controlled. SETTING: University research laboratory. PARTICIPANTS: Pigs. INTERVENTIONS: After anesthetization, in 5 pigs (25 to 30 kg), the left anterior descending coronary artery was isolated and its resting flow measured by a perivascular-Doppler flowmeter. Left ventricular pressure and first time derivative of left ventricular pressure were measured. The laser-Doppler probe needle (type N) (Model ALF-21, Transonic Systems, Inc, Ithaca, NY) was inserted 2 to 3 mm into the wall of the left ventricle, parallel to the coronary artery. All 5 pigs were subjected to 0 (control), 50% , 75%, and 100% constriction of the left anterior coronary artery. MEASUREMENTS AND MAIN RESULTS: Measurements by radio-active microspheres correlated poorly with those by laser-Doppler and extremely poorly with those by perivascular Doppler flowmeter. For percent change from baseline in the constricted arterial zone, radioactive measurements correlated well with those by laser-Doppler but not those by Doppler flowmeter. Also, radioactive measurements of percent change in flow in the circumflex (nonconstricted) zone and laser-Doppler measurements in the constricted arterial zone did not correlate well. CONCLUSIONS: Laser-Doppler can be recommended for experimental research to monitor local flow. These measurements may relate to change in regional flow during normal perfusion and hypoperfusion. Before the laser-Doppler perfusion monitor can be used clinically, tissue trauma from the 0.55-mm needle needs to be evaluated. PMID- 8725421 TI - Detection of cerebral malperfusion in chronic type "A" aortic dissection: the importance of bilateral arterial monitoring. PMID- 8725422 TI - Sudden death caused by retrograde aortic dissection during repair of a descending thoracic aneurysm. PMID- 8725423 TI - Cardiopulmonary bypass in a post-lung-transplant patient. PMID- 8725424 TI - Problems in the management of the airway during anesthesia for bilateral sequential lung transplantation performed without cardiopulmonary bypass. PMID- 8725425 TI - Anesthesia for living-related (lobar) lung transplantation. PMID- 8725426 TI - Intraoperative transesophageal echocardiographic evaluation of mitral regurgitation. PMID- 8725427 TI - Near-infrared spectroscopy: theory and applications. AB - In conclusion, NIRS appears to offer both a new monitoring modality and new information about cerebral oxygenation. Technical problems in the application of this technology persist, most notably determination of pathlength and the volume of tissue interrogated. Those familiar with the history of pulse oximetry will recall that although Millikan developed an ear oximeter in 1947, it was not until Aoyagi combined recognition of the pulse signal with spectroscopy in the 1970s that oximetry was transformed into a clinically applicable monitor. In much the same way, NIRS may find the same tremendous usefulness as a noninvasive monitor of cerebral oxygen utilization, pending resolution of the remaining technical problems. PMID- 8725428 TI - Case 2--1996. Anesthetic management of a patient with hemoglobin SS disease and mitral insufficiency for mitral valve repair. PMID- 8725429 TI - Pro: intrapleural anesthesia is useful for thoracic analgesia. PMID- 8725430 TI - Con: unreliable benefit after thoracotomy--epidural is a better choice. PMID- 8725431 TI - Dilated coronary sinus on prebypass transesophageal echocardiography. PMID- 8725432 TI - Atrioventricular sequential pacing using transesophageal atrial pacing in combination with temporary external DDD pacemaker. PMID- 8725433 TI - Continuous cardiac output measurement by thermodeprivation is unsatisfactory in cardiac surgical patients. PMID- 8725434 TI - Disappearance of signs of coarctation during dissection in a neonate--an unusual phenomenon. PMID- 8725435 TI - [The quantitative morphology of the cells in the basal layer of the multilayer epithelium in the mouse cornea and in a human cancerous lung tumor]. AB - The following categories of cells are present in the epithelium basal layer: "elongated", "transitional", "narrow", "circular", and "oval", corresponding to peaks 1, 2, A, B, and C. Peak 1 cells with the ellipticity 0.5774 have the highest specific weight. Upon maturation, these cells become "transitional" cells and mitotic, or "elongated", cells. During proliferation, some "elongated" cells mature to become more differentiated "narrow" cells. The "elongated" cells are characterized by synchronization. When 30% of the cells in the population become "elongated", 5% of them enter mitosis. Transition of 5% of the "elongated" cells to mitosis occurs only when the number of peak B cells becomes equal to that of peak A cells. When the "elongated" cells amount to 30% of the population, transition of peak B cells into peak A cells ceases. In the tumor tissue of the human lung, there are the same series and peaks of cell distribution as in the normal epithelium. There is the same functional relationship between the "elongated" cells and peak B and peak A cells as in the normal epithelium, when transition of 4-5% of "elongated" cells to mitosis is possible only upon leveling of peaks B and A. When the relative number of "elongated" cells reaches 30%, transition of peak B cells into peak A cells also ceases. PMID- 8725436 TI - [The development of the thymus in rat postnatal ontogeny]. AB - The development of thymus was studied histologically and morphometrically with an account of the seasonal factor and sexual differences during early postnatal (1-5 weeks) ontogenesis in rats. The sexual differences were more pronounced in the adrenalectomized animals. Morphogenesis of the Hassall's corpuscles during accidental thymus involution was described in more detail. Anticipating growth of the thymus medullar substance was found, as compared with the cortex, and its leading role in organogenesis, especially in 1- to 3-week-old animals, was shown. During this period, the mitotic indices of the medullar cells exceeded those of the cambial thymocytes thrice and those of the cortical thymocytes nine and five times in 1-week and 2-week old animals, respectively. In the adrenalectomized animals, the increase in the thymus cortex mass was greater in males, despite sharply decreased, as compared with females, mitotic activity of the thymocytes. The low mitotic indices of the thymocytes in the adrenalectomized animals and positive correlation of the thymus-adrenal gland mass in growing rats suggest an ambivalent effect of glucocorticoids (stimulating and inhibitory with reference to their blood content) on lymphopoiesis in the thymus cortex. Lymphopoiesis is regulated not only by the thymic or extrathymic endocrine factors, but also by contact inhibition of cell division upon achievement of a critical level of the specific density of thymocytes in the organ. Unequal distribution of mitoses in the medullar substance and in the cortex along the lobe parameter and segmentary location of mastocytes in the interlobar connective tissue suggest that a part of the lobe is an elementary structural unit of the thymus, rather than the entire lobe. PMID- 8725437 TI - [The effect of benzine and formaldehyde on the prenatal development of rats with induced iron trace-element disorder]. AB - Iron-deficiency anemia induced in the maternal organism markedly enhances embryotoxic and teratogenic effects of the studied pollutants: gasoline and formaldehyde. The prenatal effects of these drugs against the background of iron deficiency state in pregnant females leads to development of the main feature of the tissue hypoxia, decompensated metabolic acidosis, in both the maternal organism and 20-day embryos. PMID- 8725438 TI - [Serotonin inhibits phorbol ester-induced oocyte maturation in the green toad (Bufo viridis)]. AB - The activator of protein kinase C phorbol-12-myristat-13-acetate (PMA) induces maturation of the definitive intact (coated with the follicular envelopes) and defolliculated oocytes of the green toad. This effect is more pronounced in case of defolliculated oocytes. The amount of matured oocytes depends on the PMA concentration in solution. Serotonin (5-HT) inhibits or blocks maturation of the oocytes induced either by progesterone or by PMA. A possible mechanism of this effect is discussed with special reference to its role in regulation of oocyte maturation. PMID- 8725439 TI - [The enzymes of juvenile hormone metabolism in Drosophila virilis ontogeny]. AB - Time-related changes in the activity of enzymes that degrade the juvenile hormone, epoxide hydrolase, and juvenile hormone esterase, was studied in two strains (101 and 147) of Drosophila virilis. The changes of the juvenile hormone titer during the pupal-imaginal development of D.virilis are determined by the juvenile hormone esterase activity, rather than epoxide hydrolase. Among the two forms of juvenile hormone esterase present in D. virilis, one is diisopropylphosphofluoridate-sensitive and active during the studied period, while the other, diisopropylphosphofluoridate-sensitive, is present only during metamorphosis and before flight of imago and is practically absent in sexually mature individuals. Interstrain differences were shown in the activity of the diisopropylphosphofluoridate-insensitive form in the D. virilis imago. It was shown that achievement of a certain level of its activity in females of strains 101 and 147 agrees quite well with the timing of oviposition. A possible role of this form of the juvenile hormone esterase in reproduction of D. virilis is discussed. PMID- 8725440 TI - [The dynamic postnatal morphogenesis of the hemato-C-cellular interrelations in the rat thyroid gland]. AB - The system of hemato-C-cellular interactions in the thyroid gland of rats at the age of 2, 4, 12, 26, and 52 weeks is characterized by similar submicroscopical organization: the typology of hemato-C-cellular complex is preserved during ontogenesis and predominant coupling of the vascular pole of C-cell to the highly effective peripheral zone of the endotheliocytes with reference to the transport is provided. Ultrastructural changes in the zones of the perifollicular hemocapillaries facing the vascular fields of C-cells reflect the dynamics of functioning of the hemato-C-cellular system at different stages of postnatal morphogenesis. PMID- 8725441 TI - [Fluctuating asymmetry as a marker of phenotypic adaptiveness (modelled on the neurological manifestations of lumbar osteochondrosis]. AB - Analysis of palm ridge count a-b was carried out in 217 males having neurological expressions of lumbar osteochondrosis and 300 control persons. Higher indices of fluctuating asymmetry have been recorded in the group of patients thus suggesting a low stability of development. It was proposed that fluctuating asymmetry can mark phenotypes weakly adapted to postnatal environmental effects. PMID- 8725442 TI - [An electron microscopic analysis of the structure of the indirect flight musculature in heterozygotes for a new Mhc gene mutation of Drosophila melanogaster]. AB - The ultrastructure of indirect flight muscle was studied in flies (imago) heterozygous for a new allele of the gene of heavy myosin chain. Analysis of the observed defects of myofibrils suggests that in the mutants completion of differentiation of the flight muscle is disturbed. PMID- 8725443 TI - [The general biological ideas of I. I. Mechnikov]. PMID- 8725444 TI - [Some patterns in formation of antibody-antigen-antibody complexes on a solid phase: experimental study and mathematical modeling]. AB - Equilibrium and kinetic constants were determined for interactions between alpha amylase from Bacillus subtilis and polyclonal antibodies (with immobilization of either reagent). The effects of desorption of immobilized molecules and intermolecular complexes on the immunochemical reaction were studied. Models of sandwich enzyme immunoassay, whereby complexes of immobilized antibody-determined antigen-labeled antibody were formed, were developed from these results. The results obtained from the model and experiments were compared. The desorption was shown to cause the hook-effect, that is, a decrease in the label binding at high concentrations of the antigen. PMID- 8725445 TI - [Hydrolyzing ability of yeast proteases in relation to protein substrates]. AB - The possibility of use of brewer's yeast as multienzymatic preparations for hydrolyzing of animal blood proteins was demonstrated. The kinetic characteristics of hydrolysis of blood proteins by activated brewer's yeast mass and neutral proteinase from Bacillus subtilis 102 and the activation energies were estimated. Brewer's yeast biomass may be used as a source of enzymes of broad substrate specificity and noncontiguous amino acids, in particular, isoleucine, to increase the biological efficiency of blood proteins by 15-30%. PMID- 8725446 TI - [Initial kinetics of milk protein hydrolysis by chymotrypsin]. AB - The kinetics of the initial stages of hydrolysis of alpha- and beta-caseins, alpha-lactalbumin, beta-lactoglobulin, and bovine serum albumin was studied quantitatively by electrophoresis. The hydrolysis rates of caseins (demasked polypeptide chains) exceeded the rates of the degradation of milk whey proteins limited by the protein globule demasking by more than two orders of magnitude. Even relatively accessible for hydrolysis globular proteins (beta-lactoglobulin and BSA) were hydrolyzed by the one-by-one mechanism. The maximum value of the ratio between the rates of the peptide chain demasking and the protein globule hydrolysis was 0.0035. PMID- 8725447 TI - [Inhibition of phagolysis in a Bacillus thuringiensis culture by chitosan]. AB - The ability of chitosan (poly-D-glucosamine) and two chitosan salts to prevent the phagolysis of Bacillus thuringiensis subsp. galleriae strain 1-97 was studied. Chitosan and its salts inhibited the productive infection caused by two nonrelated bacteriophages 1-97A and 1-97B and suppressed the culture lysis upon spontaneous prophage induction. The efficiency of inhibition depended on the chitosan concentration, medium composition, and bacteriophage type. PMID- 8725448 TI - [Effect of Bacillus intermedius RNAase on growth of a Saccharomyces cerevisiae culture]. AB - The effects of RNase from Bacillus intermedius on proliferation of Saccharomyces cerevisiae were studied. The enzyme (0.01 microgram/ml) stimulated the yeast cell budding. This effect was dose-dependent and required an appropriate physiological stage of the growing culture cells. RNase produced maximal effects when added to exponentially growing cultures. Analysis of the age structure of the population showed that exogenous RNase stimulated the cell cycle at a stage preceding the initiation of DNA synthesis and budding of single yeast cells and cells occurring at the budding stage III. RNase did not decrease the buoyancy and osmotic sensitivity of baker's yeast. PMID- 8725449 TI - Liver resection by ultrasonic dissection and intraoperative ultrasonography. AB - Ultrasonic dissection (USD) and intraoperative ultrasonography (IOUS) have shown encouraging results in a retrospective analysis of 109 patients with benign or malignant liver disease. Of 109 patients assessed between 1980 and 1993, 84 were resected: 27 by finger fracture technique (FFT) and 57 by USD. Hospital mortality was 4.8% (4/84) and 30-day mortality was 6.0% (5/84). Overall morbidity was 48.8% (41/84) and liver related morbidity (hepatic bleeding, sepsis, and bile leak) was 34.5% (29/84); of the 29 patients, 5 required re-operation. Liver complications occurred in 12/27 (44.4%) in the FFT group as opposed to 17/57 (29.8%) in the USD group. The incidence of postoperative hepatic bleeding was significantly less by USD than by FFT (p = 0.03). As well, intraoperative blood loss (p = 0.01) number of intraoperative blood units used (p = 0.002), and postoperative length of stay (p = 0.009) have been significantly reduced by USD. IOUS was used on 64 patients. Not only has it improved the sensitivity (99%) and specificity (98%) for detection of hepatic neoplasms, it has also helped increase the precision and accuracy of anatomical tumour localization. As a result, 11/64 patients (17.2%) had their preoperative plans changed: 8 were abandoned and 3 were revised. In summary, USD has significantly reduced intraoperative blood loss and hence reduced the number of intraoperative transfusions, incidence of postoperative complications and postoperative length of stay. IOUS should be routinely employed in patients undergoing liver resection since it provides critical information that could obviate oncologically useless resections. PMID- 8725450 TI - Acute cholecystitis in aged patients. AB - The aim of this study is the analysis of the results in 62 patients over 70 years of age with acute cholecystitis treated in our Department from 1970 to 1990. The clinical picture in 47 patients was mild and in 15 severe. In 14 cases (10 calculous, 4 acalculous) the acute cholecystitis subsided with antibiotics (Group A). In 48 more cases (45 calculous, 3 acalculous) following 1-3 days conservative treatment, operation was undertaken. Besides acute cholystitis there was gangrene of gallbladder in 10, choledocholithiasis in 7 and choloperitoneum without perforation in 7 cases. Cholecystostomy in 25, cholecystectomy in 15 and cholecystectomy with exploration of the bill duct in 8 cases was performed (Group B). There was one death in group A and 3 deaths in group B. The hospital stay was 20 days. In conclusion the clinical findings in acute cholecystitis in the aged are usually mild. In the case of failure of medical treatment, after 2-3 days emergency surgery should be performed. PMID- 8725451 TI - Subtotal cholecystectomy. AB - Subtotal cholecystectomy has been carried out in 34 patients from 1972 to 1992. In the same period 1620 total cholecystectomies were performed. The indications were severe inflammation and/or severe fibrosis in 31 patients, and Mirizzi syndrome type 1 in 3 patients. The morbidity was insignificant, but one patient died, due to severe sepsis. In follow up studies ranging from 6 months to 9 years, there was one patient with retained stones in the common bile duct. No other post cholecystectomy sequelae were noticed in the remaining 32 patients. Subtotal cholecystectomy is a safe, feasible and definitive operation in patients for whom the standard operation could be dangerous. This operation is less burdensome to the patient, and is accompanied by fewer complications than ordinary cholecystostomy. PMID- 8725452 TI - Hanging noncalculous gallbladder. AB - The removal of acalculous and not acutely inflamed gall-bladder in patients with typical biliary pain remains a questionable procedure. This study was conducted to present our experience. In the period 1982-90, 1089 cases of calculous and acalculous gallbladder disease were treated in our clinic. In this period, 27 patients were subjected to cholecystectomy because of an acalculous, non inflamed gallbladder which was elongated lying in an abnormal position with a long cystic duct. The mean duration of symptoms supportive of cholelithiasis, was 5 years. Oral cholecystogram and ultrasonography led to the diagnosis and other causes of chronic abdominal pain were excluded. There were 13 lumbar, 9 pelvic and 5 iliac gallbladders, with poor function in 20 of them. During cholecystectomy, the organ was invested by peritoneum and suspended in 7 cases from a mesentery. On pathological examination mild chronic inflammation was reported in 19 cases and minimal changes in 8. The minimum follow up was one year and the maximum 9 years. Complete relief of symptoms was achieved in all the cases. In conclusion, cholecystectomy should be offered in these symptomatic "hanging" gallbladders. PMID- 8725453 TI - An easier technique for end to end pancreaticojejunostomy. AB - Breakdown of the pancreaticoenterostomy is responsible for a number of complications and for the high mortality associated with pancreaticoduodenectomy. Although in recent years the postoperative mortality has dropped to less than 10% and in some to less than 5%, pancreatic fistula remains the most common and troublesome complication. Various procedures, such as duct ligation or occlusion, resection of the pancreatic stump or pancreaticogastrostomy, have been proposed to treat the pancreatic stump when it is considered unsuitable for jejunal anastomosis. A little trick permitted us to perform 41 consecutive end to end pancreaticojejunostomies, irrespective of the conditions of the pancreatic stump, with only 3 pancreatic fistulas (7%) and without fistula related deaths. PMID- 8725454 TI - Prospective randomized trial comparing endoscopic sphincterotomy followed by surgery with surgery alone in good risk patients with choledocholithiasis. AB - BACKGROUND: Role of endoscopic sphincterotomy (ES) in high risk patients with choledocholithiasis is established but its role in good risk patients is unclear. DESIGN: A prospective randomized trial of endoscopic sphincterotomy followed by surgery (ES + S) versus surgery alone (SA) in good risk patients with choledocholithiasis. SETTING: A tertiary level referral hospital in north India; July 1991 to October 1993. PATIENTS AND METHODS: Thirty three out of 60 patients with choledocholithiasis were found suitable for randomization--16 were randomised to ES + S group and 17 to SA group. RESULTS: Common bile duct clearance was achieved in 11/13 (85%) patients in ES + S group and in 13/15 (87%) in SA group. Major complications occurred in 4/13 (31%) patients in ES + S group and 3/16 (19%) patients in SA group. These differences were not statistically significant, but patients in ES + S group were exposed to morbidity twice, procedure related morbidity of ES being 23%. No significant differences were observed in hospital stay and cost of treatment. CONCLUSIONS: Results of this trial do not support use of precholecystectomy ES in good risk patients with choledocholithiasis, since it did not offer any advantage over surgery alone. PMID- 8725455 TI - 'Latent' portal hypertension in benign biliary obstruction. AB - A prospective study was undertaken to evaluate the changes in portal venous pressure in patients with benign biliary obstruction (BBO) but without overt clinical, endoscopic or radiological evidence of portal hypertension. Portal venous pressure was measured at laparotomy in 20 patients (10 each with either benign biliary stricture or choledocholithiasis) before and after biliary decompression. Pressure was found to be on the high side in seven patients (> 25 cm of saline in three patients and > 30 cm of saline in four). The mean fall of pressure was 3.4 cm of saline after biliary decompression. No correlation could, however, be found between portal venous pressure and duration of biliary obstruction, serum bilirubin or bile duct pressure. Liver histology showed mild to moderate cholestatic changes but maintained portal architecture in all. Benign biliary obstruction may therefore, lead to elevation of portal pressure, even though the patient may not necessarily have any clinical, endoscopic or radiological manifestations of portal hypertension. The pathogenesis of this 'latent' portal hypertension is probably multifactorial. If biliary obstruction is left untreated the development of overt portal hypertension may become a possibility in the future. PMID- 8725456 TI - Effects of endothelin-1 on hepatic blood flow. AB - Endothelin-1 belongs to a family of potent vasoconstrictors, recently isolated from endothelial cells. Endothelin-1 has a variety of hepatic effects and hepatic clearance from the circulation is important. Elevated plasma concentrations of Endothelin-1 are found after orthotopic liver transplantation and in cirrhosis with ascites. This study in piglets on hepatic bloodflow was designed to compare differences in effects between central venous and intraportal injection of endothelin-1, and to evaluate effects of repeated injections. Central venous injection of endothelin-1 caused a larger reduction in portal vein flow, while intraportal injection caused a larger increase in portal vein pressure. Repeated injections resulted in a reduction in portal vein flow and an increase in portal vein vascular resistance. PMID- 8725457 TI - The concentration of trace elements in human lithogenic bile. PMID- 8725458 TI - Unusual tumors causing extrahepatic portal venous obstruction. AB - Extrahepatic portal vein obstruction has been reported to be associated with tumors of liver, bile ducts and pancreas. We report two cases, one with gastric leiomyosarcoma and another with Non Hodgkin's lymphoma, complicated by portal vein block and presenting with gastric variceal bleeding. Portal vein block in both cases was due to direct vascular infiltration. Development of portal hypertension posed difficulties in management. PMID- 8725459 TI - Hepatic segmentectomy on primary liver cancer with situs inversus totalis. AB - We present the first case treated by hepatic segmentectomy in a 69-year-old woman with primary liver cancer and situs inversus totalis. The situs inversus did not cause any technical problems during the operation, which was conducted under guidance of intraoperative ultrasonography. PMID- 8725460 TI - Gallbladder bilharziasis. AB - A 30 years old man presented with symptoms of Bilharziasis. Ultrasound showed gallstones in the gallbladder which was removed laproscopically, heavy bilharzial infection was detected in the gallbladder tissue. The first case in Saudi Arabia is reported. A review of the world literatures since 1966 about the subject is presented and different aspects of the gallbladder schistosomiasis are discussed. PMID- 8725461 TI - Treatment of large liver cyst evaluated with CA 19-9 in the cystic fluid. AB - A middle-aged woman was admitted with a diagnosis of liver cysts. The patient was symptomatic and was treated by injecting absolute ethanol into the largest cyst every week, but the secretion from the cyst persisted. The patient was then treated by absolute ethanol injection every day with good results. CA 19-9 was measured in the cystic fluid. The secretion was related to CA 19-9 activity. Thus, for patients with symptoms from a liver cyst, the injection of ethanol every day can be effective. CA 19-9 level in the contents of the liver cyst was an important factor in assessing the effect of ethanol injection on the liver cyst. PMID- 8725462 TI - Successful control of external biliary fistula by using SMS 201-995 in a child. AB - Somatostatin is a potent inhibitory hormone, it's synthetic analogue is more potent and has a prolonged action. It has a wide range of actions in the gastrointestinal system; among which is an anticholeritic action on bile secretion. The use of Sandostatin in the management of a case of complicated biliary fistula is reported; it controlled the fistula reducing its daily output from 200 ml to less than 5 ml per day. PMID- 8725463 TI - Prophylactic sclerotherapy: yes or no! PMID- 8725464 TI - Techniques of inflow occlusion for liver resection. PMID- 8725465 TI - Alcohol elimination and simulator performance of male and female aviators: a preliminary report. AB - OBJECTIVES: In this preliminary study of alcohol effects on aviators' flight simulator performance, we addressed some methodological issues regarding possible gender-related differences in response to alcohol. METHODS: Subjects were 11 male and 12 female general aviation pilots, ages 21-40. Subjects received 8 h of training before they were tested with alcohol. On the alcohol test day they were tested before drinking, while intoxicated (target BAC of 0.08%), and 8 h after drinking. RESULTS: The average, observed peak BAC readings for men and women were within 0.003% of each other. We observed faster disappearance rates for women such that women reached the FAA cutoff of 0.04% approximately 1 h before men, on average. Compared to predrink performance, there was a significant decrement in simulator performance during acute intoxication, but not 8 h after drinking. There were no significant gender differences in performance before or after drinking alcohol. Slower rates of alcohol elimination were associated with larger performance changes 8 h after drinking. This is the first report to our knowledge suggesting a possible relation between alcohol elimination rate and change in performance after drinking alcohol. CONCLUSIONS: A 12.5% dose reduction for women appears to be adequate for achieving comparable peak BAC's for male and female groups. Future studies using measures of circadian rhythmicity in conjunction with pharmacokinetic and performance measures could potentially shed light on differences in subjects' acute and delayed responses to alcohol. PMID- 8725466 TI - High performance supine flight assessment using the NAWC dynamic flight simulator. AB - INTRODUCTION: To determine the feasibility of high-performance supine flight, the USAF Canopy Escape Module (65 degree seat-back-angle) was modified and deployed into the Naval Air Warfare Center Dynamic Flight Simulator. METHODS: Seven male subjects performed a flight syllabus consisting of instrument flight maneuvers (high-G level turns, ILS task, vertical S-2, Half Cuban Eight). Subjects were trained and performed under both 1 g (static) and dynamic conditions in supine and upright postures. Data were assessed to determine the effects of motion (G) and seat position. A weighted objective performance grading scheme was devised based on the ability to achieve specified flight parameters. RESULTS: This paper presents the analysis of the first two tasks. During the turns, motion effects did not effect the ability to maintain specified G loads when supine, whereas there were significant differences when upright. Also, there were lower error rates under static, as compared to dynamic, conditions. Even though subjects demonstrated lower error rates during dynamic turns when upright, there were few significant differences referable to body position. Overall, subjects with the most flight experience performed better upright than supinated, while a naive subject flew significantly better supine. Notably, one subject did experience a G induced loss of consciousness (G-LOC) and an almost LOC event during a supinated high-G turn. Body position or motion effects did not significantly alter ILS task performance. CONCLUSIONS: High performance supine flight is feasible and additional study is warranted. Important and unanticipated performance-related decrements were discovered only through the use of dynamic flight simulation. PMID- 8725467 TI - External ocular hyperemia: a quantifiable indicator of spacecraft air quality. AB - BACKGROUND: Eye irritation consistently ranks as a top astronaut complaint but is difficult to measure. Exposure to internal air pollution hypothetically disrupts the eye's tear film, thereby exposing the crewmembers' conjunctivae to the irritating effects of the recirculated, contaminant-laden atmosphere of the space vehicle. Causes elude engineers and toxicologists, who report that measured irritants remain below established Spacecraft Maximum Allowable Concentrations. Lack of objective ocular endpoints stymies efforts to identify etiologies. HYPOTHESIS: Computers offer a practical means of analyzing ocular hyperemia in space. METHODS: We use computer analysis to quantify redness and blood vessels of digitized images of bulbar conjunctivae in near real time. Custom software masks artifacts, lids and lashes for each photographic or telemedicine ocular image, Algorithms then generate semi-independent measurements of hyperemia. Computed difference scores between 34 pairs of images were compared with subjective difference scores as voted on by a panel of ophthalmology residents. RESULTS: Objective data were reliably extracted from ocular images and significantly correlated (r = 0.583, p < 0.05) with subjective scores. CONCLUSIONS: This ground based methodology generates accurate and reliable ocular endpoint data without mass, volume, or power penalty. To assist in identifying and eliminating onboard ocular irritants, these objective data can be regressed against independent variables such as mission elapsed time, subjective astronaut complaints, levels of chemical and electromagnetic contaminants, nephthelometric and barothermal data. As missions lengthen, sensitive tools such as hyperemia quantification will become increasingly important for assessing and optimizing spacecraft environments. PMID- 8725468 TI - A comparison between phosphors for aviator's night vision imaging system. AB - BACKGROUND: The visual display on night vision goggles (NVG's) is green and isochromatic (P22 phosphor). Future systems are expected to use a P43 phosphor which has a narrower visible spectrum and is yellowish green, while the P22 is deeper green. In transitioning to the P43, some NVG's may have P22 and P43 phosphors paired in the same NVG. The purpose of this study was to evaluate the suitability of the P43 phosphor and the effect of mixing phosphors in the same NVG. METHODS: We tested three systems: one with P22 phosphors in both tubes (P22), one with P43 in both tubes (P43), and one with P22 in the right and P43 in the left tube (mixed). Visual acuity (VA), contrast sensitivity (CS), flicker sensitivity, and dynamic CS were measured in six subjects with measures repeated across the three systems (P22, P43 and mixed). RESULTS: There was no difference between systems in VA or CS across a range of simulated night sky conditions. There also was no difference between systems in sensitivity to flicker. Performance on dynamic CS was slightly better with the P43 display, which may relate to the faster decay time of this phosphor. CONCLUSIONS: These results provide no contraindication for using the P43 phosphor in NVG's (paired or unpaired), but it would be prudent to minimize mixing of phosphors in the same NVG. Additional factors that may affect performance with different color displays are discussed. PMID- 8725469 TI - The epidemiology of aircraft fire in commuter and air taxi crashes. AB - BACKGROUND: Aircraft fire is recognized as an important risk factor for occupant fatality in aviation crashes, but its epidemiology has not been adequately studied. OBJECTIVE: This study examines factors related to the occurrence of aircraft fire, ignition sources, and factors influencing rescue and firefighting in commuter and air taxi crashes. METHODS: Commuter and air taxi crashes in 1983 88 that involved aircraft fire were analyzed using National Transportation Safety Board (NTSB) data, including NTSB supplemental forms on fire and on rescue and firefighting. RESULTS: Aircraft fire was recorded in 17% of the 888 commuter and air taxi crashes. Factors associated with increased likelihood of fire in these crashes were non-airport location (adjusted relative risk 2.7, 95% confidence interval 1.8-4.1), twin-engine aircraft (RR 2.5, 95% Cl 1.6-3.9), and nighttime (RR 1.7, 95% Cl 1.1-2.5). Fire was more common in Beech 18's than other twin engine airplanes. The majority (59%) of fires occurred during impact sequence, and an additional 17% occurred after the aircraft came to rest. The engine was identified as the ignition source in 59% of the crashes, followed by hot surface (18%), short circuit (9%), and sparks (9%). Fire sensing and extinguishing systems were installed in only 19% of the aircraft. Rescue and firefighting efforts were hampered most commonly by terrain (48%), weather (47%), and darkness (38%). CONCLUSIONS: Typically induced by impact and started at the engine, aircraft fire is most likely when a crash occurs at night, in a non-airport location, and in instrument meteorological conditions. These high-risk circumstances also often thwart rescue and firefighting efforts. PMID- 8725470 TI - Hypercapnia lowers the shivering threshold and increases core cooling rate in humans. AB - BACKGROUND: Hypercapnia, which may be encountered during diving operations or by patients under general anesthesia with spontaneous respiration, alters the basic thermoregulatory responses of animals and humans. In cold-exposed animals, 3-10% inspired CO2 impairs thermal homeostasis by attenuating shivering and promoting heat loss through peripheral vasodilation. Experimental results with humans are equivocal. PURPOSE: To determine the effects of hypercapnia on warm and cold thermoregulatory responses and core cooling rates during mild cold stress, we examined the core temperature (TCO) thresholds for sweating, vasoconstriction and shivering, as well as core cooling rates of eight subjects immersed in 28 degrees C water under hypercapnic conditions. METHODS: On two separate days, subjects exercised on an underwater cycle ergometer to elevate TCO above the sweating threshold. They then rested and cooled until they shivered vigorously. Subjects inspired humidified room air during the control trial and 4% CO2/20.9% O2/balance N2 during the hypercapnia trial. RESULTS: Hypercapnia lowered the threshold for shivering by 0.13 degrees C and increased the core cooling rate by 0.35 degrees C.h-1 (25%). Minute ventilation was approximately 12.0 L.min-1 throughout cooling during control and increased from 25.3 to 28.7 L.min-1 during hypercapnia. CONCLUSIONS: Therefore, hypercapnia enhances the core cooling rate during mild cold stress. This may be attributed in part to a delay in shivering onset, as well as increased respiratory heat loss during hypercapnic hyperventilation. PMID- 8725471 TI - EGb 761 in control of acute mountain sickness and vascular reactivity to cold exposure. AB - METHOD: We recruited 44 subjects to participate in a study of the preventive effect of Ginko biloba extract (EGb 761) on acute mountain sickness (AMS) and vasomotor changes of the extremities during a Himalayan expedition. After giving their written informed consent, the subjects were randomized to two groups. One group received 160 mg of EGb 761 per day in two divided doses and the other group received placebo. Assessment was based on the course of the Environmental Symptom Questionnaire (ESQ) score and the cold gradient measured by photoplethysmography. RESULTS: The prophylactic efficacy of treatment with EGb 761 was clearly demonstrated in this study. In terms of factor 1 (AMS-Cerebral), no subject in the EGb 761 group developed acute mountain sickness versus 40.9% of subjects in the placebo group; this difference was very significant (p < or = 1.4 x 10(-3)). In terms of factor 2 (AMS-Respiratory), 3 subjects (13.6%) in the EGb 761 group developed acute mountain sickness versus 18 (81.8%) in the placebo group; this difference was very significant (p = 1.2 x 10(-5)). CONCLUSION: Due to its multiple pharmacological actions, EGb 761 provides an interesting response to the prevention of mountain sickness for moderate altitude (5400 m) with gradual exposure. It also decreased vasomotor disorders of the extremities, as demonstrated by plethysmography (p < 10(-8)) and a specific questionnaire (p < 10(-9)). PMID- 8725472 TI - Influence of granisetron on thermoregulation during exercise in the heat. AB - BACKGROUND: A NATO project group has an interest in selecting an antiemetic agent that not only is effective in the prevention of emesis induced by chemical agents or radiation exposure but also has minimal, if any, side effects. Granisetron is the second candidate drug of a class of selective serotonin antagonists that has been shown to be an effective antiemetic agent for patients receiving radiation or chemotherapy treatment. The present study was designed to evaluate whether a single 2-mg oral dose of granisetron influenced temperature regulation during exercise in a hot and relatively dry environment. HYPOTHESIS: Based on our previous findings with the other candidate drug, ondanseton, we hypothesized that granisetron would not influence temperature regulation. METHODS: Nine unacclimatized males performed a drug and placebo trial in a double-blind manner. The sessions involved walking on a treadmill at 4.8 km.h-1 with a 2% elevation for a maximum of 3 h at 40 degrees C and 30% relative humidity while wearing combat clothing. RESULTS: Granisetron was associated with a small (0.2 degrees C) but significant elevation in mean skin temperature at the beginning and after 2 h of exercise. However, there was no difference between trials for the 1.6 degrees C increase in rectal temperature. Also, body heat gain (406 +/- 97 and 407 +/- 103 k) for the placebo and drug trial, respectively) and whole body sweat rates (0.72 +/- 0.10 and 0.73 +/- 0.10 kg.h-1 for the placebo and granisetron trial, respectively) were not different. Tolerance times also were not different for the placebo (157.4 +/- 16.7 min) and drug (159.4 +/- 20.4 min) sessions. CONCLUSIONS: For the environmental conditions used in this investigation, we would accept the null hypothesis that a single 2 mg oral dose of granisetron does not influence temperature regulation during exercise. PMID- 8725473 TI - Central and peripheral noradrenergic responses to 14 days of spaceflight (SLS-2) or hindlimb suspension in rats. AB - INTRODUCTION: The purposes of this work were to assess the influence of microgravity on the central and peripheral noradrenergic activity to reevaluate SLS-1 mission findings and to compare it with that of simulated microgravity in rats. METHODS: The norepinephrine (NE) contents of the brainstem cell groups (A1, A2, A5, and A6) and organs (heart and kidneys) involved in blood pressure regulation were determined in rats after a 14-d spaceflight (SLS-2 with animals sacrificed 6 h after landing) and after a 14-d hindlimb suspension followed with 6 h of recovery. RESULTS: After SLS-2 spaceflight, NE contents were not significantly different between flight and ground-based rats either in A1 (5.2 +/ 0.5 vs. 5.7 +/- 0.4 pmol/structure), rostral A2 (12.1 +/- 0.5 vs. 11.1 +/- 0.9 pmol/structure), caudal A2 (3.2 +/- 0.6 vs. 4.3 +/- 0.5 pmol/structure) and A5 (4.4 +/- 0.4 vs. 4.3 +/- 0.5 pmol/structure) nuclei or in cardiac atria (98.6 +/- 7.5 vs. 83.4 +/- 8.9 pmol.mg-1 protein), ventricles (38.3 +/- 2.2 vs. 44.1 +/- 2.8 pmol.mg-1 protein) and kidneys (13.4 +/- 0.8 vs. 17.7 +/- 1.5 pmol.mg-1 protein). NE content was unchanged in A6 nucleus after SLS-2 comparing with control rats (respectively 4.1 +/- 0.3 vs. 4.5 +/- 0.5 pmol/structure), while it was depleted after SLS-1 mission (2.9 +/- 0.3 vs. 8.8 +/- 0.7 pmol/structure, p < 0.001) probably in relation with the stressful conditions on return to Earth. Similarly, no alterations between suspended and control rats were noted in central and peripheral NE contents after 14 d of suspension and after 6 h of recovery, whereas NE turnover studies evidenced large changes in the activities on structures on suspension and on recovery. CONCLUSION: These results suggest that only NE turnover determination will provide information about the role of the sympathetic system in the cardiovascular deconditioning. This raises the problem of the necessity to experiment inflight (injections, sacrifice) in order to avoid the recovery effects of the few hours following the landing. PMID- 8725474 TI - Swimming behavior of fish during short periods of weightlessness. AB - BACKGROUND: Birds and fish show tumbling and spinning movements when subjected to short periods of weightlessness during parabolic flight. The reason for this behavior is not clear. HYPOTHESIS: The semicircular canal system is a rotation detecting device; however, it seems that linear accelerations have an influence, too. Microgravity induces rotatory sensations which leads to a compensatory behavior (e.g., rotatory movements). METHODS: The swimming behavior of goldfish was studied with the fish in four different conditions: normal fish (group I); fish with one eye recently removed (group II); fish with both eyes recently removed (group III); and fish with both eyes removed 10 months previously (group IV). Further, a group of naturally blind (e.g., not surgically treated) cavefish (group V) were involved in the study also. All procedures conformed to the guiding principles as required in the Dutch Law on Care and Use of Animals. RESULTS: Three main different patterns of abnormal swimming behavior could be observed: tumbling (pitch), corkscrew movements (pitch and roll), and spinning movements (roll). NF did not shown any special swimming pattern. One-eyed fish (group II): mostly corkscrew movements (62%). Blind fish (group III): a mixture of the three movements (17, 22 and 25%). Blind fish (group IV): mostly spinning movements (20%). Cavefish (group V): tumbling (21%), corkscrew movements (12%) and spinning movements (58%). CONCLUSION: Vision is the dominant cue, explaining the behavior of normal goldfish. When vision is absent, the fish relies on vestibular information with respect to orientation. The swimming behavior is presumably caused by an attempt to compensate rotatory illusions. As all movements were shown in the planes of the vertical canals, we conclude that these canals play a dominant role when fish are deprived from proper otolith information. PMID- 8725475 TI - Perception of the medical risk of spaceflight. AB - We conducted an opinion survey to improve the characterization of medical risk during spaceflight, using a questionnaire designed to elicit space medicine experts' perceptions of the probability, health effect, and mission impact of selected medical events occurring during spaceflight missions of 30-90 d. This questionnaire was directed toward those events about which little data currently exist, therefore medical events that have occurred during spaceflights with some frequency, such as space motion sickness, were excluded from the questionnaire. The questionnaire was mailed to 99 clinical and research professionals involved with NASA medical programs; 65 responses were returned, of which 60 could be analyzed. The experts rated skin disorders as the most likely to occur, but which would have little effect on mission completion or astronaut health. Circulatory diseases were rated as having the lowest probability of occurrence, but the highest effect on the mission or on a crewmember's health. The results of this survey will be combined with data from analogous populations and existing astronaut health data to establish a data set to support decisions about allocation of health care resources. PMID- 8725476 TI - Back pain in aircrew--an initial survey. AB - BACKGROUND: In the early 1990's staff at a Royal Naval Air Station formed the view that pilots and observers of the Navy Lynx helicopter fleet appeared to have a higher incidence of back pain, to be seeking medical assistance more frequently and to be spending more time medically "unfit to fly" when compared to their counterparts in other helicopters. HYPOTHESIS: To quantify the incidence of low back pain in helicopter aircrew by use of a questionnaire. METHODS: A questionnaire was used to establish the prevalence and nature of back pain in Naval aircrew (161) compared to a control group of randomly selected non-flying military personnel (310). RESULTS: Results show that aircrew (82%) experience significantly more back pain than controls (52%) (p < 0.01) though the nature of pain was similar in both groups. Significantly more aircrew (73%) than controls (49%) (p < 0.01) reported a previous history of back pain and report pain in shoulders (19%), midback (42%), lower back (72%) and across the buttocks (12%). Ergonomic factors were thought to be causative. Of aircrew surveyed, 11 (8%) had been issued with personal lumbar supports which resulted in major subjective improvements in flight-related symptoms. CONCLUSIONS: Helicopter aircrew have a higher incidence of back pain. Crew station design was identified as a major contributor to the prevalence of back pain while personal supports reduced the levels of discomfort. While these findings are not new, no significant steps have been taken to redesign the crew station. It is recommended that personal supports are provided to all aircrew and that a more detailed study of crew station ergonomics is indicated following a more comprehensive study of prevalence of back pain in Royal Navy aircrew. PMID- 8725477 TI - Effect of low altitude on glycolytic key enzymes in red blood cells. AB - METHOD: The ATP level and the activities of hexokinase (HK), phosphofructokinase 1 (PFK-1) and pyruvate kinase (PK) in red blood cells (RBC) were measured in 50 school students 6-12 yr of age in the Jordan Valley (JV) which is the lowest region below sea level in the world (low altitude: 390 m below sea level, hyperoxic and with an average daily value of 796 mmHg barometric pressure) and in 50 school students of the same age group in Irbid region (normal altitude: 600 m above sea level and with an average daily value of 600 mmHg barometric pressure). The same parameters were measured also in 40 school students at sea level. RESULTS: A significant decrease in HK and PFK-1 activities and an increase in ATP level in the low altitude region, while no significant change in PK activity in JV-group when compared to Irbid and to the sea level control groups. Possible explanations will be discussed to interpret these observations. PMID- 8725478 TI - U.S. Naval helicopter mishaps: cockpit egress problems. AB - INTRODUCTION: A number of potential problems are associated with egress from the helicopter cockpit in the post-crash phase of a mishap. The purpose of this study was to identify the egress problems experienced by pilots in U.S. Navy/Marine Corp helicopter cockpits, and to propose a priority of preventive interventions. METHOD: Using Naval Safety Center data, we reviewed 1980-94 Class A helicopter mishap narratives. We identified and categorized cockpit egress problems reported by pilots. The data are described in terms of problem category, helicopter type, crash terrain, and time of day. RESULTS: Of the 210 survivable mishaps, 289 egress problems were reported in 128 mishaps, 61% involved aircrew factors, 16% environmental factors, 12% were related to helicopter factors, and 11% to cockpit factors. Of the 128 mishaps, 67.5% occurred during daytime, 32.5% at night, 64% in overwater crashes, 26% over land, and 10% over flight decks. The most significant, but uncommon, injuries involved the "stroking seat." CONCLUSIONS: Egress hazards can be minimized by the implementation of more intensive underwater egress training, crashworthy fuel systems, better design of cockpit exits and hatches, better restraint systems, better crashworthy seats, underwater visual aids, and more streamlined personal equipment. Engineering validation is needed before committing to suggested interventions. PMID- 8725479 TI - A review of Canadian Forces Search and Rescue Technician medical training and operations, 1990-93. AB - This study examined whether the 8 weeks of initial medical training and 9 d re certification every 3 yr given to Canadian Forces (CF) Search and Rescue Technicians (SAR Techs) was satisfactory. The course content was compared with 272 held medical case documents for the period 1990-93, inclusive. This practical medical care data showed a predominance of trauma rescue cases: 35% were life threatening conditions and 65% were non-life threatening conditions. They ranged from trauma, chest pain, abdominal pain, hypothermia, diabetic insulin overdose to stroke and gynecological bleeding. Of the life-threatening cases, 32% needed advanced treatment skills and 15% of the non-life threatening cases needed advanced treatment skills. It was concluded that the content of the initial and re-certification medical training was satisfactory as long as immediate transport to a specialist medical center was possible. PMID- 8725480 TI - Aeromedical certification of a pilot with bi-lateral prosthetic hands--a case report. AB - This case report involved a 47-yr-old male pilot, with 20,000 h of total flight time, who was involved in an auto accident and suffered a traumatic amputation of both of his hands, as well as head injuries with temporary cognitive deficits. He underwent rehabilitation and was fitted with Otto Bock myoelectrically-controlled upper extremity (right and left) prostheses. In the past, the FAA had medically certified a total of 17 pilots (3 Second-Class and 14 Third-Class) with unilateral amputated hands (8-left and 9-right). This was the first case involving a bilateral hand amputee. This report describes the aeromedical certification procedure used to determine this pilot's fitness for the issuance of a Third Class Medical Certificate. PMID- 8725481 TI - Treatment of motion sickness. PMID- 8725482 TI - Can cysteine direct tyrosine in signal transduction for environment-oriented gene control? AB - Signals are transduced from the cell surface to the nucleus through phosphorylation and dephosphorylation chain reactions of cellular proteins at tyrosine and serine/threonine. Recent evidence suggests that the signal generated through the protein modification at cysteine by oxidation/reduction crosstalks to the protein phosphorylation/dephosphorylation-linked one. I propose that the cysteine-oriented signal potentially directs the tyrosine-oriented one and this mechanism underlies the environment-oriented control of internal signaling for gene expression. PMID- 8725483 TI - A comparison of transurethral resection of the prostate and medical treatment for the patient with moderate symptoms of benign prostatic hyperplasia. AB - Transurethral resection of the prostate is the most common surgical treatment for benign prostatic hyperplasia. We conducted a prospective randomized clinical trial to compare this surgery with medical treatment in men with moderate symptoms of benign prostatic hyperplasia. Of 98 men over the age of 54 years who were screened between June 1993 and June 1995, 53 were studied (25 in the surgery group and 28 in the medication group). Patients' symptoms and the degree to which they were bothered by urinary difficulties were measured with standardized questionnaires and medical evaluations. The men randomly assigned to the surgery group underwent surgery within 2 weeks after the assignment. Surgery was not associated with an impotence or urinary incontinence. The follow-up period was 1 year. Surgery was significantly associated with improvement in residual urinary volume and peak flow rate; and also in the scores for urinary difficulties, sexual performance and interference with activities of daily living (P < 0.001 for all comparisons). We concluded that for patients with moderate symptoms of benign prostatic hyperplasia, surgery is more effective than medication in improving genitourinary symptoms and enhancing the quality of life. Thus, medication as treatment should be reserved for patients who are less bothered by urinary difficulty or who wish to delay surgery. PMID- 8725484 TI - 18 years of conformation radiotherapy at Nagoya University Hospital. AB - Conformation radiotherapy is one of the best techniques for minimizing the radiation dose absorbed by the surrounding normal tissue while delivering a high dose to a cancerous target area. The cases of all patients who underwent external irradiation at Nagoya University Hospital from 1975 to 1992 were reviewed. A total of 5740 patients with 6179 lesions were irradiated during this time, and 3795 treatment plans involved radical intended irradiation. Of the 5740 patients, 1017 had head and neck cancer, 982 had cervical cancer, 506 had lung cancer, 439 had primary brain tumors, 308 had esophageal cancer, 1213 had metastatic tumors, and 1275 had other types of tumors. The total number of treatment plans per year decreased from 442 in 1975 to 292 in 1992. Likewise, the percentage of conformation radiotherapy performed in all patients decreased from 29.4% (130/442) in 1975 to 8.6% (25/292) in 1992. It occupied 14.5% (982/6179) of all intended plans, and 20% (775/3795) of radical treatment plans. The conformation technique was used in cases of cervical cancer (72%), esophageal cancer (65%) and primary brain tumors (25%). Boost Conformation radiotherapy represented 2% of all treatment planning and 29% of the conformation radiotherapy. Boost Conformation radiotherapy has recently become more popular and now represents more than 50% of conformation radiotherapy. With respect to cases of cervical cancer, the rates of local recurrence and late complications in cases treated by conformation radiotherapy were lower than in cases treated by two parallel opposed radiotherapy. PMID- 8725485 TI - The incidence of antisperm antibodies in patients with seminal tract obstructions. AB - Antisperm antibodies are one of the important factors in male infertility caused by vasal obstructions. To investigate the incidence of serum antisperm antibodies in patients with various kinds of seminal tract obstructions, we retrospectively analyzed the sera of 60 patients using the indirect immunobead test (IBT) to find serum antisperm antibodies. Immunoglobulin (Ig)G and IgA class antisperm antibodies were positive in 55% and 18% of those patients with a vasal obstruction caused by inguinal herniorrhaphy and in 60% and 20% of vasectomized patients respectively; whereas these antibodies were positive in 13% and 0% of those patients with an epididymal obstruction of unknown etiology and in 8% and 3% of those patients with congenital bilateral absence of the vas deferens (CBAVD). The incidence of antisperm antibodies was significantly higher in patients with a vasal obstruction than in those with an epididymal obstruction (p < 0.001). PMID- 8725486 TI - Does epididymal length in men with congenital bilateral absence of the vas deferens have a correlation with the fertilization rate of epididymal sperm retrieved by micropuncture technique? AB - To investigate whether the variable length of the epididymis in men with congenital bilateral absence of the vas deferens (CBAVD) might have a correlation with fertilization and pregnancy rates in in vitro fertilization (IVF) treatment, we conducted a retrospective study involving a total of 60 CBAVD patients. All patients in this study had epididymal micropuncture in conjunction with perivascular nerve stimulation as part of the IVF program at the Nagoya University Hospital Reproduction Center. The patients were classified into 3 groups: group I consisted of patients having only a proximal portion of the caput epididymidis with a length of between 0.5 and 1.9 cm, group II consisted of patients with the caput and a portion of the corpus epididymidis with a length of between 2.0 and 4.0 cm, and group III consisted of patients with the caput, corpus and cauda of the epididymis measuring more than 4.0 cm. There were no differences in the sperm count, progression and normal morphology among the 3 groups; however, the motility was progressively higher in patients with a longer epididymis (12% in group 1, 18% in group II and 31% in group III). It was evident that group III showed the highest fertilization and pregnancy rate per patient (23% and 28%, respectively) among the 3 groups. This study shows that epididymal sperm from CBAVD patients with a longer epididymis have a higher IVF rate than CBAVD patients with a shorter epididymis. PMID- 8725487 TI - A study of the bone morphogenetic protein derived from bovine demineralized dentin matrix. AB - We tried to extract and purify bone morphogenetic protein (BMP) from bovine demineralized dentin matrix (DDM). Crude dentin BMP (d-BMP) was extracted from DDM in 3 sequential steps. Purification of crude d-BMP was carried out by liquid chromatography. The molecular weight and the isoelectric point of the purified d BMP were determined by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) and electrofocusing. All fractioned samples were bioassayed in the thigh muscle pouches of AKR strain mice to test their ability to induce new bone formation. A sequence of 3 extraction steps worked effectively to obtain crude d BMP. The purified d-BMP was shown to be homogeneous on high performance liquid chromatography (HPLC) and SDS-PAGE. The molecular weight and the pI were 25 kDa and 6.5, respectively. The amino acid composition was different from that of known bone-derived BMP. The purified d-BMP induced new bone formation in the thigh muscle pouches. The molecular weight, pI and amino acid composition were different from those of bone-derived BMP. PMID- 8725488 TI - Thermochemotherapy for cancer of the tongue using magnetic induction hyperthermia (implant heating system: IHS). AB - A 55-year-old patient with cancer of the tongue (T2N0M0) was treated by thermochemotherapy using interstitial magnetic induction hyperthermia (Implant Heating System: IHS). The patient received 2 courses of hyperthermia, each of which was 45 minutes long. At the same time, the patient received 2 courses of chemotherapy, which included intra-arterial infusion of 100 mg of cisplatin (CDDP) and 25 mg of peplomycin (PEP). The patient showed complete response (CR) to this therapy. To date, 1.5 years after completion of treatment, the patient has shown no recurrence. This therapy, which makes surgery and radiotherapy unnecessary, is promising, because it is expected to improve the quality of life (QOL) of cancer patients. PMID- 8725489 TI - Oxygen intake efficiency slope: a new index of cardiorespiratory functional reserve derived from the relationship between oxygen consumption and minute ventilation during incremental exercise. AB - We investigated the usefulness of the oxygen intake efficiency slope (OIES) as a submaximal measure of cardiorespiratory functional reserve. OIES was derived from the relationship between oxygen consumption (VO2; ml/min) and minute ventilation (VE; l/min) during incremental exercise, which was determined by the following equation: VO2 = a logVE + b, where "a" represents OIES, which shows the effectiveness of ventilation. Maximal oxygen consumption (VO2max) is effort dependent. There is no standard submaximal measurement of cardiorespiratory reserve that provides generally acceptable results. Exercise tests were performed by 17 normal volunteers on an ergometer using a symptom-limited Ramp protocol. Expired gas was continuously analyzed. OIES was calculated using the first 75%, 90%, and 100% of exercise data. We also determined the following submaximal parameters: the ventilatory anaerobic threshold (VAT), the slope of the minute ventilation-carbon dioxide production relationship (VE-VCO2 slope), and the extrapolated maximal oxygen consumption (EMOC). We analyzed the relationship between OIES, other submaximal parameters and VO2max, and examined the effects of submaximal exercise on OIES. The correlation coefficient of the logarithmic curve fitting model was 0.991 +/- 0.006. OIES and VO2max were significantly correlated (r = 0.966, p < 0.0001). The correlation between OIES and VO2max was stronger than the correlation between VO2max and VAT, the VE-VCO2 slope and EMOC. OIES values for 100% and 90% of exercise were identical; OIES for 75% of exercise was slightly lower (3%). Our results suggested that OIES may provide an objective, effort-independent estimation of cardiorespiratory functional reserve. PMID- 8725490 TI - Disorientation of animals in microgravity. AB - Disorientation responses of animals exposed to microgravity produced by parabolic aircraft flights and also in-space experiments were reviewed. Disoriented postures in floating are largely species-dependent. Reflexive lowered tone of gravity-bearing extensor muscles by labyrinthectomy is not seen in mammals (hamster and monkey) and frog, though dorsiflexion of the neck and the trunk is not so remarkable in hamster and monkey as in frog. In weightlessness, fundamental vestibular reflexes may be affected (righting reflex in cat), but coordinated performance can be easily compensated by visual function (mouse, monkey and turtle). In normal birds and fish who can move three-dimensionally in their environments, exposure to parabolic flight microgravity induces irregular tumbling with the eyes open and regular looping with the eyes closed, although the loop direction is the opposite in these two animals; backward (inside) in pigeon and forward (outside) in fish. Most recently, however, it was found that normal fish (goldfish) tumbled backward when observed in prolonged microgravity in space, suggesting that microgravity effects on fish in aircraft-flight parabolas would differ from those in space. Sensory conflicts in normal fish diminish remarkably during 3-4 days of space microgravity, and the cerebellum may be involved in the recovery, lending support to the sensory conflict theory for the genesis of space motion sickness. PMID- 8725491 TI - Circadian rhythm sleep disorders: a brief review with special reference to long term follow-up. AB - This article reviews the hygiene of new sleep disorders such as delayed sleep phase syndrome (DSPS) and non-24-hour sleep-wake syndrome which are classified as the circadian rhythm sleep disorders in the International Classification of Sleep Disorders (ICSD) published in 1990. It was clear in this study that these sleep schedule disturbances developed by preference in adolescence and youth, and the prevalence of DSPS in this generation was presumed to be 0.25-0.4%. Patients with certain disorders, such as entrainment to external time cues, were treated with non-pharmacological interventions and/or pharmacological agents. These therapies were effective in some cases when each was administered alone, but their effectiveness in other cases was not evident until two or more of therapies were used in combination. The prognosis of these two syndromes was significantly positive, particularly in the adolescent cases, suggesting the benefits of early discovery and treatment. PMID- 8725492 TI - X-linked recessive bulbospinal neuronopathy (SBMA). AB - X-linked recessive bulbospinal neuronopathy (SBMA) is an adult onset motor neuronopathy with androgen receptor (AR) gene mutation of expanded CAG repeat size in the first exon. The size of CAG repeats in the AR gene is one of the determinant factors of the severity and progression rate of SBMA phenotypes, but the meiotic and somatic instability of CAG repeats is far more stable as compared with other diseases caused by trinucleotide repeat expansions such as HD, DRPLA, MJD and SCA1. Several evidences suggest that aberrant transcriptional activity of androgen through mutant AR is related to the pathogenic mechanism of this disease. PMID- 8725493 TI - A case of unilateral giant hydronephrosis with renal insufficiency. AB - Giant hydronephrosis, which is a symptomless enlargement of the abdomen, is an extremely rare condition in adults, with most cases being discovered in young-to middle-aged people. We report a case of adult giant hydronephrosis accompanied by renal insufficiency, in which surgical repair was not performed because of the patient's poor cardiac function. Nephrostomy was performed and was followed with replacement of the nephrostomy tube every 2 weeks. For diagnostic purposes, ultrasonography and computerized tomography were preferable. A supplementary antegrade and retrograde pyelography was valuable in locating the stenosis. PMID- 8725494 TI - A case of sex reversal syndrome with sex-determining region (XX male). AB - We examined a 32-year-old man with a 4-year history of infertility. The man's sex life, male hair pattern, and penis were normal, and he had no history of erection problems. Left and right testicular volumes were 2 ml and 3 ml, respectively. Semen analysis showed no sperm. The endocrine panel revealed increased serum luteinizing hormone and follicle-stimulating hormone levels, and a normal serum testosterone level. A testicular biopsy demonstrated that both Leydig cell and Sertoli cell hyperplasia were present, and that no germ cells were found in the tubules. A chromosome analysis done on the peripheral blood lymphocytes revealed a karyotype of 46, XX. We identified the sex-determining region, Y, by polymerase chain reaction using Y-specific probes in this patient. The diagnosis was XX male. PMID- 8725495 TI - Distribution of neuronal cytoplasmic inclusions in multiple system atrophy. AB - Neuronal cytoplasmic inclusions (NCIs) were demonstrated by means of silver staining (Gallyas staining) in the central nervous systems of 18 deceased patients with multiple system atrophy (MSA) - 6 with olivopontocerebellar atrophy (OPCA), 6 with striatonigral degeneration (SND) and 6 with Shy-Drager syndrome (SDS). We observed NCIs in the cerebral cortex, putamen, pons, medulla oblongata and spinal cord, and especially in the putamen and pons of all cases with MSA. No NCIs were observed in the cerebellum and midbrain. The findings were common to all 3 subtypes of MSA. NCIs were not present in patients with other neurodegenerative disorders and non-neurological disorders. Our findings indicate that NCIs represent a special neuronal alteration characteristic of MSA and support the theory that OPCA, SND and SDS represent manifestations of a single condition i.e. MSA. PMID- 8725496 TI - Effects of varicocelectomy on testis volume and semen parameters in adolescents: a randomized prospective study. AB - Because the treatment of a varicocele in adolescents remains controversial, we undertook a prospective study in 51 male adolescents, aged 15-21 years, who were referred for a varicocele. Twenty-nine patients were treated with high retroperitoneal ligation of the left spermatic vein, and 22 were untreated. Eighteen healthy adolescent volunteers without a varicocele were also assessed. Testicular volume was measured using an orchiometer, and semen analyses were carried out at referral and after 1 year of follow-up. In addition, serum luteinizing hormone (LH), follicle-stimulating hormone (FSH) and testosterone levels were estimated by radioimmunoassay. Both patient groups had significantly smaller testis volumes than the controls at the outset. In the follow-up, the treated patients had testis volumes similar to those of the controls. Both testes increased significantly in volume after treatment. Although semen parameters were comparable in all groups initially, sperm concentration increased significantly after treatment of the varicocele. We concluded that varicocele treatment in adolescents leads to increased testis volume and a higher sperm concentration; however, whether early treatment will improve testicular function remains to be elucidated. PMID- 8725497 TI - Cell cycle-dependent localization of tissue inhibitor of metalloproteinases-1 immunoreactivity in cultured human gingival fibroblasts. AB - Localization of the tissue inhibitor of metalloproteinases-1 (TIMP-1) was investigated in cultured human gingival fibroblasts by immunohistochemistry. TIMP 1 immunostaining was observed in the cytoplasm of a majority of cells, and in the nucleus of some cells. Depletion of fetal calf serum (FCS) from the culture medium reduced the density of immunoreactive TIMP-1 in the nucleus more remarkably than in the cytoplasm, and the following FCS-induced cell growth was accompanied by a recovery of immuno-reactive TIMP-1 in both the nucleus and the cytoplasm. The frequency of the nuclear TIMP-1-positive cells changed in line with that of PCNA-positive cells and was always much higher than that of S-phase cells, which were estimated by a pulse labelled 5-Bromo-2' deoxyuridine (BrdU). These results suggest a localization of TIMP-1 or a related substance in the nucleus of proliferating human fibroblasts, and its depletion from the nucleus due to an arrest of cell growth. PMID- 8725498 TI - A case of primary glucocorticoid resistance. AB - A 79-year-old woman developed hypokalemia and metabolic alkalosis after breast cancer surgery. She was suspected of having primary glucocorticoid resistance on the basis of high plasma ACTH and serum cortisol levels without the features of Cushing's syndrome. To clarify the end-organ resistance to cortisol, we characterized the glucocorticoid receptors (GR) in cultured skin fibroblasts from the patient. The GRs in whole cell assays decreased binding affinity (Kd = 11.1 +/- 0.6 nM) and the number of binding sites for [3H]dexamethasone (binding capacity was 15,600 +/- 1,255 sites per cell). These results strongly suggest that our patient had primary glucocorticoid resistance caused both by a decreased number of GRs and a reduction in the affinity of GRs to cortisol. PMID- 8725499 TI - Assessment of the effects of aging and medication on salivary gland function in patients with xerostomia using 99mTC-scintigraphy. AB - To examine the effect of aging and medication on xerostomia, the salivary gland function was evaluated in 20 patients with xerostomia using 99mTc-scintigraphy and the measurement of unstimulated whole saliva (USWS). All of the patients showed USWS volume of less than 2ml/10min. The patients were divided into 2 subgroups based on age (under 65 and 65 and older) and medication status (patients who were on medication which reduced salivary secretion and patients who were not on such medication). The scintigraphic results, such as the maximum radioisotope (RI) count, RI secretion velocity and the volume of USWS, were compared between the subgroups. The maximum RI count and the RI secretory velocity in the submandibular gland and the volume of USWS revealed significantly different functional disturbances between relatively younger patients (under 65) and older patients (65 and older). There was no difference when the scintigraphic results and the volume of USWS measurements in medicated patients were compared with the results of similar tests performed on non-medicated patients. When the medicated and non-medicated groups were separated by age, an increase in age still diminished the volume of USWS in medicated patients. This result might be related to an organic change in the submandibular gland in older patients which was suggested by the scintigraphic results. PMID- 8725500 TI - An evaluation of the biological effects of three different modes of magnetic fields on cultured mammalian cells. AB - The biological effects of static magnetic fields, and their combined effects with ionizing radiation, were studied using a cultured mammalian cell line (FM3A). The three different modes of magnetic fields evaluated in this report were the 0.3 Tesla (T) field with a gradient of 0.3T/m, the 0.7T field with a gradient of 0.7T/m and the 6.34T field with no gradient. Exposure to the 0.3T and 0.7T fields had no effect on cell survival. Exposure to the 6.34T field decreased cell survival. Survival curves showing the combined effect of the 0.3T and 0.7T fields with radiation had a smaller mean lethal dose (D37) value. The survival curve of the 6.34T field was influenced by the interval between magnetic exposure and ionizing irradiation. When the interval was 6 or 12 h, the survival curve showing the combined effect of the 6.34T field had smaller D37 and quasithreshold dose (Dq) values, indicating the potentiation of the radiation effect. Flow cytometric analysis indicated that exposure to the 0.3T and 0.7T fields showed no change and that exposure to the 6.34T field showed an increase in the percentage of G1 phase cells. Our conclusions were as follows: 1) magnetic fields decreased the colony forming abilities of cultured mammalian cells; 2) magnetic fields can affect the cell cycle; 3) a stronger magnetic field strength does not always have stronger biological effects and 4) the gradient of a magnetic field may be an important factor when combined with ionizing radiation. Despite the foregoing analysis, the biological effects of magnetic fields on mammalian cells remains a complex phenomena. PMID- 8725501 TI - Leiomyosarcoma of the maxillary gingiva: a case report. AB - An unusual case of leiomyosarcoma (LMS) of the maxillary gingiva is discussed here; this case presents a unique pattern of tumor growth and a long period between initial discovery and correct pathological diagnosis. The tumor was incompletely resected twice by a private dentist over a period of 3 years, with a clinical diagnosis of epulis, no pathological examination was conducted during this period. When it was finally removed, the tumor was very large (50 x 35 x 12 mm in size and 18 g in weight), consisting of an easily hemorrhagic mass originating in the gingival mucosa with the growth pattern of a polyp. Following an extensive surgical excision and a unilateral radical neck dissection, the patient has been free of LMS for 8 years. In light of this case, we strongly emphasize the importance of conducting a pathological examination, even though clinical examination seems to indicate a diagnosis of epulis or granulation. In this way, the presence of LMS can be ascertained in a timely manner with better prognosis for treatment and recovery. PMID- 8725502 TI - The clinical and histopathological effects of combined chemotherapy using cisplatin and peplomycin to treat cancer of the tongue. AB - Combined chemotherapy (PP therapy) using cisplatin (CDDP) and peplomycin (PEP) was performed as induction chemotherapy for 31 patients with cancer of the tongue, and the clinical and histopathological effects were investigated. As the primary clinical effect, complete response (CR) was observed in three cases, partial response (PR) in 20 cases, minor response (MR) in six cases and no change (NC) in two cases, with a clinical response rate of 74.2%. The histopathological effects of the chemotherapy in the following cases showed these histological Grades: 0 or I in four cases, IIa in 12 cases, IIb in eight cases, III in six cases and IV in one case. Fifteen of the 31 patients who received PP therapy showed a histological Grade of IIb or better, representing 48.4% of the histopathological response rate. With regard to the mode of invasion of the tumor, the histopathological response rate was 90.0% in patients with invasive Grades 1 and 2, 41.7% in those with invasive Grade 3 and 16.7% in those with invasive Grade 4C. There were no patients with invasive Grade 4D in whom the therapy was histopathologically effective. In other words, the histopathological effect significantly decreased as the invasive Grade increased. With regard to the relationship between clinical effects and histopathological effects, there was one CR patient who showed a histological Grade of IIa. Thus, it is noteworthy that clinical effects were not necessarily consistent with post-chemotherapeutic histopathological effects. PMID- 8725503 TI - Immune functions and immunopathology of the mucosa of the upper respiratory pathways. AB - The specific defence of airway mucosae depends primarily on secretory immunity. The B cells involved are initially stimulated in organized mucosa-associated lymphoid tissue, apparently including the tonsils and adenoid. From these inductive sites, memory cells migrate to secretory effector sites where they differentiate terminally to immunoglobulin (Ig)-producing plasma cells. Locally produced Ig consists mainly of J chain-containing dimers and larger polymers of IgA (pIgA) that are selectively transported through glandular cells by an epithelial receptor called secretory component or the pIg receptor. IgG can participate in immune exclusion because it reaches the secretions by passive diffusion. However, its proinflammatory properties render IgG antibodies of local immunopathological importance when elimination of penetrating antigens is unsuccessful. T helper (Th) cells activated in this process may by a Th2 cytokine profile promote persistent inflammation with extravasation and priming of eosinophils. This development appears to be part of the late-phase allergic reaction, perhaps initially driven by interleukin-4 (IL-4) released from mast cells that are subjected to IgE-mediated activation, and subsequently also by Th2 cell activation. Eosinophils are potentially tissue-damaging, particularly after priming with IL-5. Various cytokines up-regulate adhesion molecules on endothelial and epithelial cells, thereby enhancing migration of eosinophils into the mucosa, and perhaps in addition causing aberrant immune regulation within the epithelium. Soluble antigens bombarding the epithelial surfaces normally seem to induce several immunosuppressive mechanisms, but mucosal homeostasis appears less patent in the airways than oral tolerance to dietary antigens operating in the gut. PMID- 8725504 TI - Eosinophils in the pathophysiology of nasal polyposis. AB - Nasal polyposis can be defined as a chronic inflammatory disease of the paranasal sinus mucosa, leading to a protrusion of benign edematous polyps from the meatus into the nasal cavities. Nasal polyps are histologically characterized by massive edema and accumulation of eosinophils. IgE-mediated allergy seems to play only a minor role in eosinophil accumulation, leaving the place for a new concept of non allergic rhinitis with eosinophilia. The central question still remains, however, why eosinophils accumulate into nasal polyposis tissue. Some initial data show that tissue structural cells, i.e. epithelial cells or fibroblasts, could produce cytokines (GM-CSF) and play a role in eosinophil accumulation (micro environmental theory). However, further studies showed, that GM-CSF was mainly produced by eosinophils themselves (autocrine theory), leading to the hypothesis of an intrinsic eosinophilic inflammatory process. Eosinophils may contribute to nasal polyp formation and growth not only through inflammation but also by exerting their effects on extracellular matrix including stimulation of collagen synthesis. Another feature associated with nasal polyposis is aspirin sensitivity. Some preliminary data indicate that eosinophils could also be involved in aspirin-sensitivity mechanisms. PMID- 8725505 TI - Effects of corticosteroid therapy in non-allergic rhinosinusitis. AB - Corticosteroids have a multifactorial effect initiated by their binding to a specific cytoplasmic glucocorticoid receptor. At the cellular level there is a reduction in the number of antigen-presenting cells, in the number and activation and T cells, in the number of epithelial mast cells, and in the number and activation of eosinophils. Steroids have a proven effect on symptoms and signs in non-allergic rhinosinusitis with eosinophilia and in nasal polyposis. Topically applied drugs, studied in many controlled trials, reduce rhinitis symptoms, improved nasal breathing, reduce the size of polyps and their recurrence, but have a poor effect on the sense of smell and no direct effect on sinus pathology. Systemic steroids, less well studied, appear to have an effect on all types of symptoms and pathology, the sense of smell included. A short course of systemic steroids is as effective as polypectomy with a snare. Individualized management of nasal polyposis and non-allergic rhinosinusitis with eosinophilia may consist of long-term topical steroids, short-term systemic steroids, or surgery, in various combinations. PMID- 8725506 TI - Three-dimensional analysis of nystagmus induced by neck vibration. AB - The role of dorsal neck proprioceptive inputs to vestibular compensation was investigated in 11 patients with unilateral vestibular dysfunction. Subjects had neither spontaneous nystagmus nor disequilibrium, indicating that they were well compensated. Vibratory stimulation to the dorsal neck produced marked nystagmus. All subjects showed horizontal component directed towards the contralateral side of the lesion. Vertical and torsional components of the nystagmus were exhibited by 10 and 7 subjects, respectively. These results indicate that the neck vibration caused a discompensation in vestibularly well compensated subjects after unilateral dysfunction. In addition, the influence of neck afferent on the vestibulo-ocular reflex pathways, not only of the horizontal, but also the vertical systems, is discussed. PMID- 8725507 TI - Diplopia from skew deviation in unilateral peripheral vestibular lesions. AB - Vertical diplopia from skew deviation is well described in brainstem lesions. The phenomenon can also result from peripheral vestibular lesions. During the past years, we have observed these ocular changes in the acute stage following unilateral vestibular neurectomy and labyrinthectomy (n = 13), as well as in series of patients suffering from idiopathic sudden unilateral peripheral vestibular or cochleo-vestibular deficit (n = 5). Diplopia from skew deviation was noted immediately following ablative vestibular procedures; in patients with idiopathic vestibular deficit, it was observed as an associated sign. In all patients, clinical evaluation revealed an acute unilateral peripheral vestibular loss, with spontaneous nystagmus toward the unaffected ear and absence of nystagmic response to caloric testing on the affected ear. Skew deviation was measured using the Hess-Weiss test, which is based on the haploscopic principle. Static visual vertical was evaluated with the original methods of vertical frame and Maddox rod techniques. Photographs were made of the ocular fundi, to measure the degree of cyclotorsion of both eyes. In our patients, we found skew deviation with hypotropia of the eye that was ipsilateral to the affected ear and conjugated cyclotorsion and tilt of the static visual vertical on the side of the affected ear. Skew deviation was the first sign to disappear within a few days; conjugated cyclotorsion and tilt of the static visual vertical persisted for weeks to months. The eye-head postural reaction, consisting of head tilt, conjugated eye cyclotorsion, skew deviation, and alteration of vertical perception directed toward the side of the lesion, is known as the Ocular Tilt Reaction (OTR). The mechanism is presumably related to a lesion of the otolithic organs and/or to changes in the afferent graviceptive pathways. In man, the OTR is often mild and unrecognized, masked by spontaneous nystagmus and marked neuro vegetative symptoms. Our observations indicate that skew deviation, as a part of the OTR, occurs in patients with sudden peripheral vestibular lesions, whether surgical or non-surgical in origin. PMID- 8725508 TI - Study of the accuracy of stance regulation. AB - In order to find a rule underlying the control of stance we recorded the center of pressure intermittently every 29 ms, first with subjects standing still for 30 s and then during shifting the center of gravity along the margin of the supporting area for 30 s to detect any movement. Subjects were 5 adult volunteers. They were examined with eyes open and closed as well as standing on one leg, closed legs, and legs 15 cm apart. The distribution of cumulated densities of the center of pressure, in a 3-D drawing, formed a very simple graph, i.e., a tower on a circle. As the difficulty of stance increased, the circle was reduced and the height of tower lowered, independently of visual or supporting condition. Since easy and difficult stances could be simulated by small and large values of SD in a regular distribution, stance may yield action oriented computation based on probability. PMID- 8725509 TI - The subjective horizontal in eccentric rotation influenced by peripheral vestibular lesion. AB - Peripheral vestibular equilibrium disorders may originate in various parts of the labyrinth or the vestibular nerve. Traditionally, the function of the lateral semicircular canals has been assessed with caloric irrigation, and sometimes falsely been interpreted as a vestibular nerve lesion. The vertical semicircular canals are not easily assessed. Caloric testing with the head in different positions is not very helpful, but the canals may pairwise be tested using specific rotational techniques. Often the otolithic organs, capable of detecting linear acceleration forces, are forgotten as a source of vertigo and dizziness. We have implemented horizontal rotatory testing with the subject seated eccentrically facing the direction of rotation as a means of assessing otolithic function. The subject experiences a lateral tilt and is instructed in darkness to put a short light bar in the position he thinks a water surface would have, which is identical to his perceived tilt. In 39 normal subjects, a theoretical tilt of 24 degrees was estimated as approximately 19.5 degrees, and the standard deviation was 6 degrees. A side difference index was proposed to be normal if below 25%. The preliminary findings in a few patients with known labyrinthine lesions are presented. We propose that otolithic as well as lateral semicircular canal functions are useful to monitor in patients with suspected peripheral equilibrium disorders. PMID- 8725510 TI - Hearing loss and vestibular dysfunction in Meniere's disease. AB - In a retrospective study of 475 patients suffering from Meniere's disease, the hearing loss, both in the low and high frequencies, and the loss of vestibular function were studied over time. There were large variations of hearing loss from case to case but, overall, the low were more affected than the high frequencies, regardless of time elapsed since onset of disease. Hearing deteriorated over time and mean values of hearing loss correlated well with the elapsed time since onset of disease. There were also large variations of post-caloric labyrinthine preponderance (lp) from case to case but, overall, lp deteriorated with time. Mean values of lp, however, did not correlate that well with elapsed time since onset of disease as the hearing loss did. Nor was there any correlation over time between level of hearing loss and lp. It is concluded that in Meniere's disease the hearing, deteriorating over time, is mainly but not necessarily worse in the low frequencies; lp, indicating loss of vestibular function, does not correlate with level of hearing loss in most cases. PMID- 8725511 TI - Receptorpharmacological models for the therapy of labyrinthine vertigo. AB - In the mammalian labyrinth, GABAA receptor subtypes are involved in the excitatory neurotransmission between the vestibular type II hair cells and the afferent neurons. Additional afferent ionophoric receptor channels, sensitive to further transmitter candidates, are discussed for both types I and II hair cells. GABA accelerates excitotoxic cell death in cortical neurons. This GABA-ergic neurotoxic action forms an appropriate pathophysiological model explaining peripheral labyrinthine disorders characterized by the spontaneous labyrinthine nystagmus and vertigo in man. A calculated GABAA receptor antagonism was envisaged in order to attenuate the pathological vestibular imbalance following one-sided GABA-accelerated vestibular neurotoxicity. Moderate allosteric blockers of the GABAA receptor channel and weak inverse agonists of the benzodiazepine binding site meet some requirements for potentially successful clinical application. The suppressing action of the suitable drugs picrotoxin and flumazenil on labyrinthine nystagmus and vertigo, tested in clinical trials, supports the hypothesis that GABAA receptors are involved in vestibular neurotransmission, even in humans. The test results promise the development of a successful vestibular receptorpharmacology in the near future. PMID- 8725512 TI - Altered behaviour of hamsters by prolonged hypergravity: adaptation to 2.5 G and re-adaptation to 1 G. AB - We studied the functional adaptation process in 40 hamsters subjected to either prolonged hypergravity to normal gravity. Subadult golden hamsters (n = 20) exposed to a hypergravity condition of 2.5 G for 6 months were tested to investigate the effect of hyper gravity on the perceptive motor skills and compared with control hamsters (n = 20). The motor coordination of the hypergravity hamsters hardly changed; locomotion was normal and swimming was possible. Equilibrium maintenance was disturbed during the first 3 months as was shown by the higher crossing time (p < 0.001) and higher fall frequency (p < 0.001) for the hypergravity group. Significant differences were also found in orientation during swimming (p = 0.007) and turning behaviour in the rotation task (p < 0.001) and in the no-rotation task (p = 0.029). After 6 months, 10 hamsters of both groups were tested for another 4 months, also the hypergravity hamsters were living at 1 G. Differences in orientation in the two groups did not change during swimming and turning behaviour during the rotation task (p = 0.026). Based on our findings, we conclude that the hamsters functionally adapted to hypergravity, which led to an altered performance of several tasks. The condition continued after 4 months of normal gravity. PMID- 8725513 TI - Influence of the extraocular muscle proprioceptors on the orientation of the vestibulo-ocular reflex. AB - In the intact brain lamb, unilateral electrolytic lesion of the medial dorso lateral portion of the semilunar ganglion containing the first order neurons of the eye muscle proprioception induced modifications of the horizontal and vertical vestibulo-ocular reflex (HVOR and VVOR) which consisted in marked alterations of the trajectories of the quick phases, while the slow phases were scarcely affected. Similar results were observed after section of the branches described by Winckler in the retrobulbar region along the extraocular muscle proprioceptive information travels. These findings extend those of previous investigations carried out in decorticate animals. PMID- 8725514 TI - An experimental technique for determining middle ear impedance. AB - A two-microphone technique was used to determine the middle ear impedance of a live subject. The procedure involved the application of standing wave tube theory and the assumption that the ear canal behaves like an homogeneous cylinder with plane acoustic wave propagation up to a certain frequency--2 kHz for the current analysis. During experimentation the subject lay on a bench with his head braced against a wooden fixture. Acoustic pressures were recorded from the ear canal by the use of a spectrum analyser and probe microphones with flexible tips. Resultant impedance curves show middle ear natural frequencies at 831 Hz and 1,970 Hz with high levels of damping. The reactive impedance curves show the influence of stiffness and ossicular mass on middle ear sound transmission. An advantage of the approach is that using features of the recorded data it is possible to calculate the effective probe tip to eardrum distance required for the calculation of the middle ear impedance. The two-microphone technique appears to be a promising tool for assessing healthy and diseased middle ear function. PMID- 8725515 TI - Is perilymphatic pressure altered in tinnitus? AB - Tinnitus is characterized by the continuous or intermittent auditory perception of various sounds (buzzing, whistling, etc.) in the absence of any external stimulus. Perilymphatic hyperpressure is one of the numerous mechanisms which could hypothetically be involved in tinnitus generation. In the present experiment, perilymphatic pressure was measured indirectly using the tympanic membrane displacement technique. Twenty-five tinnitus patients were investigated at 10, 15 and 20 dB above the acoustic reflex threshold with ipsilateral stimulation. The variables Vi (inward tympanic displacement), Vm (mean tympanic displacement) and their variations according to stimulus level were compared between tinnitus sufferers and age-matched or hearing-matched controls. Tympanic displacement was measured in sitting and supine positions so as to evaluate cochlear aqueduct patency. No systemic changes in response occurred in tinnitus patients, except at a high stimulation level, perhaps due to hearing impairment. PMID- 8725516 TI - The effect of methylprednisolone on acoustic trauma. AB - The effect of methylprednisolone (mPSL) upon acoustic trauma was studied using albino guinea pigs which were exposed to 2 kHz pure tone of 110, 115 or 120 dB SPL for 10 min. After the exposure to intense sound, mPSL of 6, 12 or 40 mg/kg was intraperitoneally given daily for 7 days and the threshold of the compound action potential (CAP) was examined on the 8th day. Compared with CAP of the control animals given physiological saline solution, no significant difference was observed in the CAP threshold shift between the mPSL group and the controls following exposures to the sound of 115 and 120 dB SPL, respectively. However, when the animals were exposed to the sound of 110 dB SPL, the CAP threshold shift was significantly smaller in the mPSL animals than in the controls. The present results indicate that mPSL possesses a therapeutic effect in cases of mild acoustic overstimulation. PMID- 8725517 TI - Ipsilateral suppression of transient evoked otoacoustic emission: role of the medial olivocochlear system. AB - Contralateral sound stimulation produces suppression of transient evoked otoacoustic emission (TEOAE), which is attributed to a reflex activation of the medial olivocochlear system. More pronounced suppression of TEOAE produced by ipsilateral masking could involve efferent-mediated effects along with effects of cochlear origin. However, this has not been investigated so far. Therefore, changes of click-evoked OAE under ipsi- and contralateral forward masking by clicks and noise-bursts were investigated in an extremely long-lasting experiment in one normal-hearing volunteer. The reduction of TEOAE under ipsilateral click to-click forward masking was maximal during the first milliseconds after masker delivery implying that predominant role of the cochlear processes in TEOAE ipsilateral suppression. Ipsilateral forward masking by noise burst revealed additional TEOAE suppression with longer latency. Its time course was similar to that of the contralateral masking effect. The latter data suggest the involvement of the medial olivocochlear system in TEOAE ipsilateral suppression. PMID- 8725518 TI - Auditory consequences of early mild hearing loss associated with otitis media. AB - Two groups of children received periodic documentation of their middle ear status by pneumatic otoscopy in the first year of life and had hearing sensitivity estimated by multiple auditory brainstem response (ABR) assessments during the same period. One group was considered otitis media (OM)-positive, with repeated bilateral episodes of OM and mild conductive hearing loss. The second group was considered OM-free with normal middle ear function bilaterally and normal hearing sensitivity in the first year of life. Children's higher-order auditory abilities were examined at 4, 6 and 9 years of age. Various measures such as speech recognition-in-competition, visual-auditory learning, sound blending, auditory memory, and masking level difference estimates were utilized to probe the children's auditory abilities. Results show that children with a first-year history of otitis media demonstrate deficits in the long term in some aspects of higher-order auditory processing. Performance appears to be associated with the mild hearing loss experienced during an important period of early development. PMID- 8725519 TI - Auditory fractal random signals: experimental data and clinical application. AB - In the mammalian primary cochlear afferents, fractals in the postsynaptic bursting behaviour triggered by a constant perisynaptic release of glutamatergic transmitter agonists have been demonstrated. In order to test the validity of fractally coded auditory signal transmission in man, frequency, intensity and temporal resolution tests were performed in cochlear implanted patients. All patients clearly recognized the fractally coded signals transmitted to the cochlear implants. These first results demonstrate evidence for fractally coded auditory signal transmission in man. PMID- 8725520 TI - Auditory plasticity in cochlear implant patients. AB - Dynamic range (DR) of cochlear implant electrodes and speech recognition scores (SRS) were chronologically studied in 23 post-lingually deaf subjects over a period of one year. DR significantly increased over one year (ANOVA, p = 0.03). When DR was compared among three groups of electrodes which were located in the inferior, ascending and superior segments of the cochlear basal turn, that in the superior segment showed the largest increase (p = 0.001), followed by those in the ascending and inferior segments. Vowel and consonant recognition scores with auditory cue were best 6 months after operation and were maintained at the level for one year. Although the time course of improvement differed between DR and SRS, a significant positive correlation was observed at 12 months after operation. Improvement of SRS could be a result of a DR increase in the superior segment of the cochlea, which should convey pitch information over the mid frequency region. PMID- 8725521 TI - Ipsilateral ABR with cochlear implant. AB - Ipsilateral ABR recording technique was developed with the MXM DIGISONIC DX10 cochlear implant, involving, firstly, setting of recording variables with regard to implant and ABR constraints, and, secondly, enhancement of recording quality by signal processing. The resulting recording quality then enabled us to characterize ABR latency, amplitude and wave reproductibility according to stimulus intensity and stimulation site (electrode stimulated). The findings agree with those of the literature on contralateral human and ipsilateral animal studies. Waves III and V amplitude increased with stimulus intensity. Waves III and V latency was insensitive to stimulus intensity or site. ABR quality diminished basally. PMID- 8725522 TI - Development of capacities of communication and socialization in young deaf children: utility of a common assessment protocol for implanted or hearing aid equipped children. AB - Pediatric cochlear implantation has attracted a growing interest among health professionals, but it has also encountered opposition on the deaf community. It is therefore incumbent upon the medical teams that carry out this procedure to furnish scientific arguments demonstrating the benefits of this technique for the age group 2 to 4 years and its innocuousness for the child's later psychological development. With this in mind, we have elaborated an assessment protocol adapted to very young deaf children, regardless of whether they have implants or hearing aids. The present report describes two tests of the protocol, chosen because they meet needs which until now have been insufficiently, or not, met. The first of these tests aims at assessing, as early as the age of two, oral and gestual production thanks to sequential images. The second test is the most original element of our psychological protocol: analysis of the child's interactional attitudes through filmed game situations. The initial results from a group of children recently implanted and from another group equipped with hearing aids are presented. These children continue to be seen at regular intervals. It is reasonable to hope that this kind of study will afford objective arguments to the debate in progress. PMID- 8725523 TI - Speech perception and speech production skills of children with multichannel cochlear implants. AB - The acquisition of speech perception and speech production skills emerges over a protracted time course in congenitally deaf children with multichannel cochlear implants (CI). Only through comprehensive, longitudinal studies can the full impact of cochlear implantation be assessed. In this study, the performance of CI users was examined longitudinally on a battery of speech perception measures and compared with subjects with profound hearing loss who used conventional hearing aids (HA). The average performance of the multichannel cochlear implant users gradually increased over time and continued to improve even after 5 years of CI use. Speech intelligibility was assessed from recordings of the subjects' elicited speech and played to panels of listeners. Intelligibility was scored in terms of percentage of words correctly understood. The average scores for subjects who had used their CI for 4 years or more exceeded 40%. PMID- 8725524 TI - Hormonal modulation of inner ear fluids. AB - In the cochlea, hormones such as antidiuretic hormone and adrenocorticosteroid hormones are supposed to modulate the endolymph osmolality acting on the labyrinthine water permeability, on the one hand, and on the Na+, K(+)-ATPase, on the other hand. To test the hypothesis that these hormones are involved in the inner ear fluids homeostasis, the electrochemical composition of cochlear fluids was studied in control Long Evans rats, Brattleboro rats that are genetically deprived of antidiuretic hormone, and in adrenalectomized Long Evans rats. The results demonstrated that: i) in Brattleboro rats, the endocochlear K gradient was absent whereas the endocochlear potential and the Cl concentration gradients were maintained; the K gradient was restored by the dDAVP administration; ii) in adrenalectomized rats, no modification of the electrochemical composition of endolymph occurred; the injection of bumetanide (10 mg/kg) induced a larger decrease of the endocochlear potential in adrenalectomized rats than in control animals. These results suggest that the cellular transport systems involved in the endolymph secretion may be altered by different hormones such as antidiuretic hormone and/or adrenocorticosteroid hormones. Nevertheless, the hormonal modulation of the inner ear fluid homeostasis remains to be further documented. PMID- 8725526 TI - Electrocochleography during experimental cochlear ischemia of the guinea pig. AB - It has been postulated that impairment of cochlear blood flow (CoBF) is one of the most important causes of hearing loss occurring during acoustic neuroma (AN) surgery. However, it remains unclear how the degree of cochlear ischemia influences the evoked responses in electrocochleography. (ECochG) which has been used for monitoring cochlear functions. In the present study, we investigated alterations in ECochG during cochlear ischemia of varying degree in the guinea pig. In order to induce cochlear ischemia, the anterior inferior cerebellar artery (AICA) was mechanically compressed via the transclival approach. The compression of AICA resulted in the reduction of CoBF in 55 out of 70 guinea pigs. A constant reduction of CoBF was maintained during the compression of AICA in 44 (63%) guinea pigs. CoBF abruptly decreased upon compressing AICA, and promptly recovered after releasing the compression. N1 and N2 in ECochG were also altered by compression. During 3-min ischemia, N1 and N2 disappeared in 36% and 41% of the cases, respectively. The residual CoBF in cases whose N1 and N2 disappeared was significantly lower than that in other cases whose N1 and N2 were sustained during 3-min ischemia. In addition, there was a tendency that the lower the residual CoBF was, the shorter the survival time of N1 and N2. In cases whose N1 and N2 did not disappear, the prolongation of N1 and N2 latencies after 3-min ischemia was positively correlated to the residual CoBF. On the basis of these results, we discuss the mechanisms underlying the changes in CoBF and ECochG during cochlear ischemia, and conclude that the degree of cochlear ischemia during AN surgery can be estimated with ECochG. PMID- 8725525 TI - Effect of neurotrophic factors on the inner ear: clinical implications. AB - Loss of auditory neurons is commonly associated with sensorineural deafness, and may result from either direct neuronal injury or be a consequence of sensory hair cell loss (i.e. loss of source of trophic factors). Developmental studies and in vitro studies of adult neurons have begun to identify growth factors important for the development, maintenance, and rescue/repair of auditory neurons. Specific neurotrophic factors have been shown to enhance the auditory neurons' ability to withstand traumatic loss of target tissue connections and toxic injury. Promising initial in vivo studies confirm that specific neurotrophins are able to support neuronal survival and promote neuronal repair in an intact animal following injury to the cochlea. Further study into unique methods and routes of growth factor delivery will provide insights into the possibility of neurotrophic growth factors to act as drugs for the treatment of injured or stressed auditory neurons. PMID- 8725527 TI - Post-traumatic survival and recovery of the auditory sensory cells in culture. AB - Following mechanical injury in organotypic cultures, auditory hair cells show the ability to survive and to initially reform their apical specializations, cuticular plates and stereocilia, but none show incorporation of tritiated thymidine, the mitotic marker. Disruption of the reticular lamina and local injury to hair cell cuticular plates induces proliferation of supporting cells. The regenerating apices of inner hair cells are wrapped by the cells of the inner spiral sulcus and the inner phalangeal cells, while those of outer hair cells are wrapped by the phalangeal processes of Deiters' cells and outer spiral sulcus cells. Some of these hair cells subsequently resurface with newly formed tops. Hair cells that lose contact with the surface of the organ remain buried--but alive--deep within the epithelium. Our study provides evidence that the mammalian organ of Corti responds to injury not by the formation of new sensory cells but by the recovery of the pre-existing postmitotic hair cells. PMID- 8725528 TI - Pantothenic acid and coenzyme A in experimental cisplatin-induced ototoxia. AB - We have observed that pantothenic acid (PA) prevents deafness induced by cisplatin (CP) in the guinea pig if both drugs are administered jointly. When deafness was previously produced, recovery was sometimes obtained after the administration of PA; so, we studied the effects of PA on cisplatian-induced ototoxia in guinea pigs, both as a prophylactic agent in healthy animals, and as a therapeutic agent in animals previously made deaf by the drug. To elucidate why PA protects the ear from the toxic effects of CP, we used coenzyme A (CoA) instead of PA-since PA is a component of CoA-to test the hypothesis that the action of PA is due to CoA. The results were practically the same in both experiences, the compound action potential of the auditory nerve (CAP) was tested and cochleas were examined by scanning electron microscopy (SEM). Our results suggest that the protective effect of PA takes place through CoA. Both substances had the same effect on CP ototoxicity, but CoA appears to be much more active, since the dose tested here was much lower than that of PA. PMID- 8725529 TI - Experimental model of vertigo induced by detached otoconia. AB - Isolated posterior semicircular canals (psc) of bull frogs were used for a model of positional vertigo. Induced ampullary nerve action potentials were recorded. When the cupula was removed and the saccular otoconia were dropped onto the cilia, excitatory, and inhibitory potentials were evoked by changing the psc positions into canal-down and canal-up, respectively. When the otoconia were allowed to stay on the cupular surface, canal-down and canal-up positions also evoked excitatory and inhibitory responses, respectively. When the otoconia were inserted in the canal and were allowed to move, changing the psc positions likewise evoked responses. However, when one end of the canal was closed, these responses disappeared, indicating the efficacy of canal plugging. PMID- 8725530 TI - Immunohistochemical and electrophysiological demonstration of substance P in human vestibular ganglion cells. AB - This study was performed in order to determine the immunohistochemical expression and distribution pattern as well as the electrophysiological actions of substance P in human vestibular ganglion cells. Substance P-like immunostaining was mainly found in the cytoplasm of small vestibular ganglion cells. In contrast, nerve fibers were not labeled. Using intracellular recording techniques, substance P produced a slow depolarization of membrane potentials accompanied by an increase in membrane resistance. Furthermore, an enhanced firing response to depolarization occurred. PMID- 8725531 TI - Effects of clofilium, a K channel blocker, on electrogenic K secretion and afferent discharge at the frog semicircular canal. A preliminary report. AB - Application of clofilium to the endolymphatic side of the isolated frog semicircular canal abolished the transepithelial potential and produced increased K and mannitol outfluxes from the lumen to the dark cells or the basolateral perilymph, with no apparent effect on active K secretion. These results suggest an increased permeability of the paracellular pathway. When applied to the perilymphatic side in the intact labyrinth, clofilium reduced the rates of quantal transmitter release (miniature EPSP frequency), an effect that might arise from a decrease in the transduction current intensity secondary to the reduced transepithelial electrochemical potential for K+. Moreover, afferent spike rates were almost completely abolished at rest as well as during mechanical stimulation. This effect together with a decreased mEPSP amplitude points to a further direct action of clofilium on the afferent postsynaptic terminal. These results suggest a multi-factorial effect of clofilium that would reduce the sensitivity of the vestibular function. PMID- 8725532 TI - Marginal shell of the anteroventral cochlear nucleus: acoustically weakly-driven and not-driven units in the unanesthetized decerebrate cat. AB - The ventral cochlear nucleus (VCN) of the cat is encapsulated by a marginal shell which is anatomically distinct from the central core of the VCN. Physiologically, little is known about the marginal shell. We report here a new finding that the marginal shell of the anteroventral cochlear nucleus (AVCN) in the unanesthetized decerebrate cat contains neural units which were weakly driven or not driven acoustically. The locations of the single units were histologically determined. Our observations also included other neural units of the marginal shell which were strongly driven acoustically; the latter results will be reported separately. The present physiological findings, together with previous anatomical findings of vestibular and somatosensory inputs to the AVCN marginal shell, suggest that some neurons of the AVCN marginal shell receive non-auditory sensory inputs besides weak auditory inputs and that these may play a role in multi sensory processing. PMID- 8725533 TI - The pars flaccida middle ear pressure and mastoid pneumatization index. AB - The degree of pars flaccida retraction and the levels of mastoid pneumatization were assessed and correlated in 388 adults with intact pars tensa. Poorly pneumatized mastoids were found to be associated with retraction of pars flaccida; the poorer the pneumatization, the deeper the retraction. Well pneumatized mastoids were usually associated with normal position of the pars flaccida. The pars flaccida was previously also shown to retract in face of ME negative pressure--and its degree can be seen to be an index of ME negative pressure. Thus, the correlation of deeper degrees of pars flaccida retractions with mastoid hypopneumatization (and vice versa) lends strength to the studies which show the mastoid pneumatic system to have a function of a passive ME pressure buffer. This observation lends further evidence as to why ears with poorly pneumatized mastoids are a priori at risk to develop complications such as SOM in adults, tympanic membrane retractions and perforations, incus necrosis or retraction pocket cholesteatoma. Ears with a large pneumatic system are hardly at such risk. PMID- 8725534 TI - PCR assay for detecting Streptococcus pneumoniae in the middle ear of children with otitis media with effusion. AB - We compared a newly developed pneumococcal polymerase chain reaction (PCR) for Streptococcus pneumoniae (Pnc) to bacterial culture in 123 middle ear effusion (MEE) samples of 123 children with otitis media with effusion (OME). For the pneumococcal PCR assay, DNA of MEE samples was purified by a QIAamp blood kit. The outer primers used amplified a 348 basepair region of the pneumolysin gene, and the inner a 208. Pnc was cultured in 14 (11%) and pneumolysin PCR was positive in 57 (46%) of the 123 MEE samples. All the culture positive samples were also PCR-positive. Both the samples with culturable Pnc and with positive pneumolysin PCR increased with shorter duration of OME and a greater number of acute otitis media during the preceding 6 months. In conclusion, pneumolysin PCR suggests pneumococcal involvement in MEE even in OMEs with no evidence of Pnc in culture, and thus offers a good diagnostic tool when a more accurate and sensitive pneumococcal diagnosis is needed. PMID- 8725535 TI - Squamous metaplasia of the middle ear epithelium. AB - This study deals with the expression of cytokeratins (Cks) in squamous cell metaplastic lesions in rat and human middle ear. In rats, squamous metaplastic lesions could be induced during chronic otitis media. The histological features of these lesions were similar to those observed in the human middle ear. Immunohistochemistry revealed that squamous cell metaplasia in both rat and human middle ear is characterised by a loss of simple epithelial cell related Cks and the appearance of Cks characteristic of stratified and cornifying epithelia. This indicates a true change in the differentiation of the middle ear epithelium. It is concluded that the Ck profile of the cholesteatoma matrix cannot be used as a variable to decide whether the origin of cholesteatomas is epidermal or metaplastic. This rat model is suitable for studying squamous cell metaplasia in relation to cholesteatoma genesis. PMID- 8725536 TI - Dynamic multivariate modelling: day care and consultation rate for acute otitis media. AB - A random sample of 2,512 children was monitored to the age of 2 years to study the effects of various risk variables on the consultation rate of acute otitis media. A total of 3,283 episodes of acute otitis media of 2,411 children with a follow-up time of at least 1 month were included in the analysis. During a 3 week period after onset of the episode, the number of unplanned visits varied between 0 (2,544 episodes; 77.5%) and 6 (2 episodes; 0.1%). The children with day care outside their home showed a diminished risk for unplanned visits during the first week. The odds ratio (OR) and 95% confidence interval (CI) for children in family day care were 0.58 and 0.39-0.8, and 0.72 and 0.47-1.11 for children in nursery day care, respectively. During the second and third weeks the results were reversed: the above odds ratios should be multiplied by interaction terms 2.19 (CI 1.41-3.42; new OR 1.27) for children in family day care and 1.57 (CI 0.97 2.54; new OR 1.13) for children in nursery day care. PMID- 8725537 TI - Interleukin-1 alpha, interleukin-1 beta and interleukin-8 gene expression in human aural cholesteatomas. AB - Bone destruction is a common characteristic feature of chronic otitis media, especially aural cholesteatoma. A number of immunohistochemical studies have suggested that interleukin-1 (IL-1) may be responsible for cholesteatomatous bone destruction. We designed this study to present the mRNA expression patterns of IL 1 alpha, IL-1 beta, and IL-8, which can induce and activate the leukocyte, the major reservoir of potent proteolytic enzymes. Total RNAs were extracted from aural cholesteatomas, external auditory canal skin (EACS), postauricular skin (PAS), and granulation tissues and transcribed into cDNAs. cDNAs were amplified by using PCR technique with primers for IL-1 alpha, IL-1 beta, IL-8, and beta actin. Amplified products were hybridized with each internal probe and the relative density was measured. In granulation tissues, the relative density of IL 1 alpha was greater than that of other tissues. The ratio of IL-1 beta and IL-8 of aural cholesteatoma was significantly higher than that of EACS and PAS. We suggest that both of IL-1 alpha and IL-1 beta may play a role in the pathological changes, and that IL-8, which is mainly produced from cholesteatomatous epithelium, may have an important role in the pathological changes of cholesteatomas. PMID- 8725538 TI - Developmental changes in folding of the human eustachian tube. AB - Histologic sections of human temporal bone with Eustachian tube attached were examined to investigate the changes that occur in folding of the Eustachian tube (ET) as it develops postnatally. Study of 45 specimens from individuals aged 3 months to 88 years of death, including 5 cases with chronic otitis media (OM) with perforation and 5 with OM with effusion, showed that the folds in the tube decreased significantly as age increased up to 20 years, after which no significant change with age was found. These results suggest that folding in the ET is important to its function in infants and children. Folding provides a relatively greater surface area for the tube, allowing for a greater abundance of ciliated cells, which would promote clearance of the middle ear through the ET. Greater folding might also support the tube's protective function by keeping the lumen narrow. PMID- 8725539 TI - Functional morphology of nasal blood vessels in humans. AB - "Secretion" and "obstruction" as predominant clinical symptoms in rhinology affect a great number of patients with disorders of the nose. According to clinical experience as well as morphological investigations, the endonasal vascular system is most likely to be involved in these functional mechanisms. In the present study, we report morphological findings on the angioarchitecture of human nasal mucosa. Meticulous investigation of the structure and ultrastructure of nasal mucosal capillaries revealed differences in the appearance of the endothelial lining. Especially the morphological feature of attenuated fenestrated endothelia in these vessels might be correlated with the functional behaviour under various physiological and pathological conditions. Inspection of the vascular wall of nasal swell bodies revealed differences in the orientation as well as the shape of muscle cells in different parts of this vascular system. The functional role of special morphological features known as muscular bolsters and intervascular muscle fibres for the swelling mechanism of the nasal mucosa is emphasized. Our results enabled us to define the muscular structures that are most probably responsible for constriction and dilatation of nasal swell bodies. PMID- 8725540 TI - Effects of viral and bacterial infection on nasal and sinus mucosa. AB - There is still controversy over the role of viruses and bacteria in rhinologic infections, especially in sinusitis. Until recently it was not fully known whether the sinuses do in fact take part in the infectious process of a common cold (viral rhinitis). CT scans show that in the vast majority of otherwise healthy volunteers with a common cold, and without a previous history of recurrent or chronic sinusitis, the sinuses are involved; there was, however, in these individuals no typical symptomatology of acute sinusitis. A viral rhinitis alone does not seem to be able to elicit a "clinical" acute otitis. Bacteria determine the clinical picture and outcome of sinusitis. There is not much controversy about the role of bacteria in acute sinusitis, S. pneumoniae, H. influenzae, and M. catarrhalis being the most frequently involved. Much more conflicting reports are published about the normal flora of the sinuses, the role of anaerobes and the microbiology of chronic sinusitis. In this paper the mechanisms of viral and bacterial infection of the nasal and sinusal mucosa are described and the results of microbiological studies in sinusitis reported by other authors and our own group are discussed. It is postulated that, although bacteria are very important in acute sinusitis, their role in chronic sinusitis is minimal, the bacteria being opportunistic colonisers. PMID- 8725541 TI - Some morphological, histological, cytological and histochemical aspects of Reinke's oedema. AB - Using morphological criteria we divided Reinke's oedema into a pale (transparent) and a livid type. The latter is more difficult to treat because of its prolonged postoperative recovery. Ten samples of pale Reinke's oedema and 10 of livid oedema were selected for cytological, histological and histochemical analysis in order to determine some of their fundamental characteristics and differences. Tissue samples were stained by hemalaun-eosin, Mallory-azan and Paf-Halmi method (specific for visualization of elastic versus collagen fibres). Cytology (May Grunwald Giemsa) of cellular elements was undertaken by imprint method of the excised oedematous tissue. Also, the silver-staining nuclear organizer regions (Ag NOR) were identified and quantified. We identified intracytoplasmic granula and silver-staining nuclear organizing regions (Ag NOR) in stromal cells of pale type Reinke's oedema. Thus, it may be supposed that smoke irritation provokes higher activity of stromal cells producing a special substance in Reinke's space resulting finally in Reinke's oedema. PMID- 8725542 TI - Cytokeratin and vimentin expression in normal epithelium and benign lesions of the vocal cords. AB - The expression of cytokeratins and vimentin was studied immunohistochemically in normal epithelium and 12 benign lesions of the vocal cord with the use of a broad panel of monoclonal antibodies against cytokeratins and vimentin. Histology showed that the various lesions contained hyperkeratotic, hyperplastic and atrophic epithelium, irrespective of their clinical appearance. Especially the Ck profile of the (hyper)keratotic lesions was changed in comparison with the native epithelium. Increased expression of the keratinization marker Ck 10 was associated with decreased expression of the stratification markers Cks 4 and 13. Expression of the basal cell marker Ck 14 and hyperproliferation-associated Cks 16 and 17 was increased in all the benign lesions, except in atrophic epithelium. These expression patterns differ from those observed in malignant epithelial lesions. The latter show a marked expression of simple cell Cks and vimentin and more pronounced expression of hyperproliferation-associated markers than the benign lesions. PMID- 8725544 TI - Histomorphological characteristics of non-metastatic lymph nodes in patients with head and neck cancer according to their site in the neck. AB - The prognostic relevance of histomorphologically defined reaction patterns of lymph nodes was demonstrated in head and neck cancer. Till today the lymph node site in the neck has not been taken into account. Therefore histomorphologically defined reaction patterns of non-metastatic lymph nodes in the neck were examined according to their site in the neck. A total of 1188 dissected neck lymph nodes of 32 previously untreated patients with carcinoma of the head and neck region were examined histologically by serial sections and classified into 6 different lymph node levels and 6 histomorphological types. The objectivity of the morphological examinations could be evaluated morphometrically by means of an interactive image analysis system (IBAS) and a more precise definition of the lymph node reaction types could be established. The results of the morphological and morphometrical classifications show a statistically highly significant relationship (p < 0.0001) between the reaction types of lymph nodes and their site in the neck. These findings allow a division of the neck in an upper anterior half with immunologically active lymph nodes and a lower caudal-dorsal half, which contains essentially less immunologically active lymph nodes. PMID- 8725543 TI - The role of FDG-PET in the preoperative assessment of N-staging in head and neck cancer. AB - This prospective study based on 48 patients showed that FDG-PET has a significantly higher sensitivity for the detection of lymph node metastases compared with palpation and it appears that FDG-PET has a similar sensitivity to CT-scanning. According to our data, FDG-PET is a highly specific method in the evaluation of neck nodes. This new imaging technique allows a tridimensional study and is easy to interpret. Therefore, FDG-PET seems to be a valuable imaging technique for the detection of cervical lymph node metastasis. PMID- 8725545 TI - Increased radiosensitivity is associated with p53 mutations in cell lines derived from oral cavity carcinoma. AB - The curability of oral cavity carcinomas, as well as of other head and neck cancers, varies remarkably especially in more advanced disease. Radiotherapy and surgery, including large operations, are currently combined, but as new radiotherapy regimens are being introduced, the need for predictive assays has increased in order to plan a suitable individual treatment for the patient. Variations in intrinsic radiation sensitivity of cancer cells cannot alone explain differences in therapy outcome, and thus additional predictive variables have to be searched. Mutations in the p53 tumor suppressor gene are found in most head and neck tumors, which has led us to study the possible association between these mutations and radiation sensitivity. We analyzed 16 cell lines from oral cavity carcinomas and found a remarkable variation in radiosensitivity (AUC 1.7 2.3 Gy and SF2 0.31-0.51). The p53 gene was mutated in 11/16 cell lines, and these cells were also significantly more sensitive than those with wildtype p53 (AUC 1.9 +/- 0.2 Gy and 2.3 +/- 0.2 Gy, respectively, p = 0.023). Structural alterations in the p53 gene were also observed in three of the relatively resistant cell lines, which indicates that not all mutations are critical in this respect. PMID- 8725546 TI - Somatostatin receptor scintigraphy and therapy of neuroendocrine (APUD) tumors of the head and neck. AB - Through the development of somatostatin scintigraphy with the labeled somatostatin analog Indium111-Octreotide, it has recently become possible to accurately diagnose primary tumors of the APUD system as well as their metastases, since these tumors usually have somatostatin receptors. Experience with this method is already available for endocrine and exocrine tumors of the gastrointestinal tract, neuroendocrine and breast tumors, small cell bronchial carcinomas and certain lymphomas. In the present study, this new diagnostic technique was used for the first time in various head and neck tumors (carcinoid of the larynx, Merkel cell tumor, glomus tumor of the carotid and glomus jugulare tumor). Concurrently, some of these tumors shown by this diagnostic method to be somatostatin receptor positive were treated using the somatostatin analog Octreotide, a therapeutic approach new for the ENT-specialty. Our initial results prove that the detection of the ENT tumors which we studied by means of receptor scintigraphy is reliable. The preliminary results of this Octreotide therapy show a growth inhibitory effect, especially for those tumors of the head and neck which are inoperable or are difficult to approach surgically. PMID- 8725547 TI - Significance of adhesion molecules as biological prognostic factor in locally advanced laryngeal squamous cell carcinomas. AB - Thirty laryngeal carcinomas were studied immunohistochemically in order to evaluate whether the expression and different distribution of adhesion molecules influence the clinical features and progression of the tumors. On the basis of clinical and pathological variables, two different groups were established: one with good and the other with poor prognosis. The patients were included in one of the two groups on the basis of prognostic factors previously studied by multivariate analysis (the validity of this choice was confirmed by the NED survival curves of the two groups). Different integrins, type I and V laminin and type IV collagen were evaluated by means of monoclonal antibodies in the tumoral specimens and in normal mucosa. Univariate statistical analysis was performed to evaluate differences between the two groups. The degree of expression and pattern of distribution were different in tumor compared with normal mucosa and significant differences were found between the good- and worst-prognosis tumors. PMID- 8725548 TI - Experimental in vitro and in vivo studies of epithelium formation on biomaterials seeded with isolated respiratory cells. AB - Extensive tracheal defects after intensive care medicine, trauma, or large resections in tumor surgery remain a major challenge in plastic and reconstructive surgery. Defects that cannot be satisfactorily treated by complicated and costly reconstructive techniques reveal a need for an alloplastic tracheal replacement. Recent experimental and clinical studies in the development of alloplastic tracheal prostheses proved that the lack of an epithelial lining on the luminal surfaces and inadequate biophysical properties and shapes of the prostheses were the main causes for failure of these prostheses. In this study a cell-seeding technique has been used. Adhesion, spreading, and differentiation of seeded mucosa cells on biomaterials in vitro were observed by scanning electron microscopy (SEM). Chemical properties and surface structure of the material influenced the differentiation process. Epithelium formation of incorporated tracheal prostheses was tested in animal experiments. Isolated respiratory cells were seeded into implanted tubular prostheses of porous polyurethane or expanded polytetrafluorethylene. Light microscopy and SEM showed the tendency of epithelium formation on the surface of the lumen. Vigorous cell layers, predominantly as multiple cell layers of squamous epithelium, were observed. Ciliated or mucus cells were not detected. It can be stated that the epithelium formation on incorporated porous implants is possible. Further studies of the stability and the differentiation process of the epithelium on such implants is needed before an introduction of tracheal replacements into the clinical practice can be considered. PMID- 8725549 TI - Long-term clinical experience with continuous ambulatory peritoneal dialysis: access-related problems. AB - Patients with end-stage renal failure can be treated with peritoneal dialysis, which is based on the capacity of the peritoneum to exchange fluid and metabolic products. To achieve this, dialysis fluid has to be instilled in the abdominal cavity through a permanent percutaneous access device. Apart from the advantages of peritoneal dialysis, severe problems are related to the access device. In this study, catheter-related morbidity and mortality are described, as found in the patient population from the University Hospital, Nijmegen, The Netherlands. The overall rates of exit-site infections and peritonitis are respectively 0.80 and 1.36 infection episodes per patient-year. Furthermore, it appeared that exit-site infections and peritonitis are the main reasons for discontinuation of dialysis and removal of the catheter. A correlation between the occurrence of peritonitis and exit-site infections was found. Also, the efficacy of the antibiotic treatment necessary to control these infectious complications is described. It is concluded that the design and the materials used to manufacture the currently used access device are the main reason for the existing morbidity in peritoneal dialysis. Therefore, more efforts should be undertaken to improve the access device, in which the design and the material used are critically considered. PMID- 8725550 TI - Characterization of a murine model of acute lung injury (ALI): a prominent role for interleukin-1. AB - This report describes a model developed to study local and systemic events that occur as a result of acute lung injury (ALI). C57BL/6J mice were injected with a single intravenous dose (2, 4, and 6 micrograms) of 12-O-tetradecanoylphorbol-13 acetate (TPA). At 1, 2, 4, 12, 24, and 48 h, after injection, plasma was collected by sinus orbital puncture, bronchoalveolar lavage (BAL) was performed and cells and fluid were collected, lungs were perfused, and pulmonary tissue was isolated and processed for histological, immunochemical, and gene expression studies. The results indicate a dose-dependent increase in animal distress and a decrease in survival. TPA induced an early systemic response, reflected as an initial decrease in numbers of peripheral blood neutrophils at 1 h, followed at 2 h by a sustained increase. There was dose- and time-dependent increase in IL-1 beta mRNA synthesis, detected using RT-PCR, and in immunoreactive IL-1 alpha produced by both tissue-fixed pulmonary cells and cells within alveolar spaces. Infiltration of neutrophils into pulmonary tissue and increased protein content in BAL fluid was detected 2 h after injection of TPA. Disruptions in pulmonary architecture accompanied by the presence of highly vacuolated macrophages within the alveolar spaces and interstitial tissue were evident after IV injection of TPA. The study shows that injection of TPA induces reproducible dose- and time dependent alterations in cell types, numbers, state of activation, and production of soluble mediators in the peripheral circulation within BAL and pulmonary tissue. Thus, this model offers a means to examine the cellular basis for the local and systemic alterations observed during ALI. PMID- 8725551 TI - Gastrointestinal sutureless anastomosis using fibrin glue: reinforcement of the sliding absorbable intraluminal nontoxic stent and development of a stent placement device. AB - Sutureless anastomosis of the gastrointestinal tract using fibrin glue and sliding absorbable intraluminal nontoxic stents (SAINTs) has two shortcomings, stent shaft breakage and the lack of a transanal insertion device (TID) for low anterior resection. Reinforcement of the sucrose base SAINT (R-SAINT) is described. Sutureless anastomosis is attempted using a stapleless mechanical stapler (SS) and used as preprototype to screen histologically and mechanically for TID anastomoses in the small intestine. Finally, a prototype absorbable head SAINT placement device (SAINT-PD) intended for TID, similar to the SS, is utilized on the small intestine. Fifty-seven Landrace pigs weighing 25-35 kg were used to perform 58 anastomoses, including the small intestine (15 manual, 19 SAINT, 11 SS, 5 R-SAINT, 6 SAINT-PD) and large intestine (2 R-SAINT). All anastomoses performed with the R-SAINT succeeded on the first attempt even if the shaft cracked. The SS technique proved impractical, but the histological screen results from 7 to 60 days did approximate those of corresponding SAINT anastomoses. The SAINT-PD demonstrated operational improvement over the SS, but the histological results were similar to both the SS and SAINT. The advantages of the R-SAINT and SAINT-PD are that they leave no foreign bodies or pressure clamping devices at the anastomostic site. Larger studies may show the R-SAINT and the SAINT-PD to be practical, new surgical tools in sutureless fibrin glue anastomosis. PMID- 8725552 TI - Development of an in situ isolated porcine liver perfusion model for tightly controlled physiologic and pharmacologic studies. AB - Several types of isolated perfused porcine liver models have been proposed for the study of hepatic assist, preservation injury, and specific physiologic or pharmacologic mechanisms. The development of a more general in situ isolated perfused model applicable to a broad range of studies is presented. This model eliminates or minimizes the shortcomings of previous models including ischemic injury prior to perfusion, limited range of vascular pressures and flows, nonphysiologic sources of portal and hepatic artery perfusion, and coupling of the liver to uncontrolled whole-body homeostatic mechanisms. Essentially the model as presented can be described as an autologous transplanted liver without preservation or ischemic injury, functioning within an adrenalectomized, cardiac output and temperature-controlled animal. Independent control of the dual hepatic vascular supply is maintained with pulsatile perfusion of the hepatic artery from the left atrium and nonpulsatile perfusion of the portal vein via the portal system. Oxygenators are not required. Hepatic vein pressure can be controlled independently of hepatic blood flow and systemic hemodynamics. Pharmacologic studies are not restricted to drugs whose termination of action is limited to hepatic metabolism because normal routes of drug redistribution, metabolism, and excretion are present. The model exhibits normal oxygen metabolism and classic control of hepatic artery resistance by portal vein blood flow. There are rather obvious significant advantages inherent in this model for tightly controlled hepatic physiologic and pharmacologic studies. PMID- 8725553 TI - Role of poloxamer 188 during recovery from ischemic spinal cord injury: a preliminary study. AB - Paraplegia following aortic surgery is not a common event. When it does occur it significantly alters the patient's outcome. Poloxamer 188 (P188) has been shown in the experimental animal to increase regional blood flow to ischemic areas. In order to investigate its protective effect during aortic cross-clamping, 23 animals were randomized to two groups (placebo n = 11, P188n = 12) and received an intravenous injection of placebo or P188 (200 mg/kg), and underwent occlusion of the thoracic aorta and both subclavian arteries for a period of 13 minutes. They were then connected to an intravenous pump delivering either placebo or P188 (250 mg/kg/hr at a rate of 0.942 ml/hour) for 48 hours. Hindlimb function was appraised, daily for 30 days, by a lesion score (0-15). Spinal cord injury was assessed by a histologic score (0-3) based on the degree of gray and white matter gliosis, number of motor neurons, and white matter myelination. Analysis of variance for repeated measures did not reveal significant difference between P188 and placebo groups (P = 0.66). Similarly, the mean histologic scores (placebo = 1.54 +/- 0.41 SE, P188 = 1.08 +/- 0.33 SE) did not differ (Wilcoxon, P = 0.43). We conclude that intravenous administration of P188 before, during, and for 48 hours after aortic cross-clamping does not prevent paraplegia or improve the long term neurologic outcome. PMID- 8725555 TI - David Elliott Rogers 1926-1994. PMID- 8725556 TI - Woman, M.D.: issues, perceptions and choices. AB - I would end with some summarizing points and questions. 1. A majority of women identify gender-related issues influencing their career choices and pathways. 2. Women are diverse in their choices and priorities, with varying balances of personal and professional life. 3. Eighty-three percent of the women responding to the survey have married, and 82% of those married have children. They thus are likely to have two sets of seriously competing responsibilities. 4. It is possible to identify especially promising women during residency or fellowship. Exploring their priorities and values with good mentors will be important to avoid short-changing academic medicine of competent women. 5. Men and women need to be SUPERCHOOSERS in making the choices right for that individual, rather than superwomen or supermen. 6. We can question whether the development and potential of women (and some men) in academic medicine could be enhanced by: a. Consideration of guidance or support for child care resources? b. Flexibility in the system (tenure clock, etc.)? c. Identification of limited full-time clinical or research opportunities for faculty with young children or unusual family demands? d. Facilitation of bridging after a period of part-time activity or re entry after limited participation? e. Consideration of part-time work at home and flexible on-site work hours for periods of special family demands? Finally, a supportive work environment is important for both men and women in medicine and is an essential priority in academic medicine. Medicine is a wonderful field. Academic medicine on my terms, to include family, has been very rewarding. Sometimes there also have been disappointments, in that I have fallen short of some of my aspirations. However, I must accept the responsibility for my choices. I owe a great debt to many of the men and women of the American Clinical and Climatological Association and of Johns Hopkins who are both friends and colleagues and especially to my husband, Dick Johns. After 42 years of marriage, I am still learning about the balance of personal and professional life. May we all be Superchoosers! PMID- 8725557 TI - The thalassemia syndromes: lessons from molecular medicines index case. AB - The thalassemia syndromes were the first of human diseases to become thoroughly examined for the underlying molecular lesions by the application of molecular genetic strategies and recombinant DNA methods. Students of thalassemia have now enjoyed over two decades of experience with this research paradigm. These experiences reveal both the awesome power and the limitations of the "reductionist, deterministic" approach of gene cloning and analysis. Incredibly precise and abundant information about the exact molecular lesions responsible for various forms of thalassemia were rapidly obtained by the use of molecular genetic approaches. The mechanisms by which these mutations deranged globin gene expression could be documented with extraordinary precision and efficiency. Precise, powerful methods for detecting disease early in fetal life were rapidly developed, made practical for field use, and disseminated widely. This resulted in a dramatic reduction in the incidence of new births of patients with homozygous beta thalassemia. These experiences demonstrate the extraordinary impact that recombinant DNA technology has upon our ability to understand disease processes, to detect disease long before its phenotypic expression is apparent, and to influence the prevalence of the abnormal alleles in the population. Experience with the antenatal diagnosis of the thalassemias also demonstrates, and should alert us to, the relative ease with which genetic information can be applied to societal and governmental initiatives to alter the reproductive behavior of individuals. While the benefits of reducing the incidence of beta thalassemia are clearcut, application of the strategies that were applied in this narrow situation to broader aspects of disease or genetic manipulation does raise concerns. The thalassemia syndromes demonstrate that genetic information does have more than a theoretical potential to have a major impact upon society. The struggles of many investigators to develop effective pharmacologic agents for the treatment of hemoglobinopathies have also revealed some of the limitations of an isolated molecular approach to the understanding of disease. The tortuous course by which a class of reagents has been identified for stimulation of HbF synthesis illustrates an important point. The application of recombinant DNA methods revealed an entirely new array of pathophysiologic facts that stimulated new hypotheses about the regulation of gene expression and opportunities to manipulate that regulation therapeutically. However, practical application and proper understanding of the molecular information were achieved only when those data were placed in the context of cell biology, tissue and organ-based clinical pathophysiology, and clinical pharmacology. Progress was possible only because of the productive interaction of talented individuals with expertise in these different fields. Our two decades of experience with the thalassemias illustrate very clearly the fact that biology and disease are extraordinarily complex, non deterministic processes. They will be understood and treated properly only if thriving centers exist within which individuals with diverse interests, expertise, and perspectives about basic science and clinical medicine can exist, interact, and have sufficient time to employ their imaginations to the fullest benefit. PMID- 8725558 TI - High prevalence of cobalamin deficiency in the elderly. AB - In summary, we have shown that there is a high prevalence of Cbl deficiency in the elderly. This observation is based on an increase in the number of low and low normal serum Cbl levels, an increase in elevated serum total homocysteine levels that correct with Cbl therapy, and an increase in elevated serum methylmalonic acid levels that also correct with Cbl therapy. These observations may be of great clinical importance since Cbl deficiency may be a common and treatable cause of vascular disease. PMID- 8725559 TI - A new voyage of discovery: transport through the hepatocyte. AB - In summary, hepatocellular membranes likely play an essential role in the binding and directed trafficking of unconjugated bilirubin, and potentially of a variety of other small hydrophobic molecules. Targeting of these substrates to the endoplasmic reticulum is determined by membrane cholesterol content, surface area and integral protein binding and enzyme activity. The rate of intracellular transport potentially may be modulated by the concentration of cytosolic binding proteins, but, at least for ligandin, this protein does not appear to function primarily as an intracellular bilirubin transporter. PMID- 8725560 TI - Angiotensin converting enzyme inhibitors--do they have an anti-ischemic action? PMID- 8725561 TI - The impact of performing a clinical trial on patient outcomes: lessons from the Emory Angioplasty vs. Surgery Trial. PMID- 8725562 TI - The role of coumadin and aspirin in atrial fibrillation. PMID- 8725563 TI - The Gordon Wilson lecture: unique interactions between viruses, neurons and the immune system. AB - The increased susceptibility of young individuals to alphavirus encephalitis is likely to be linked to the ease with which immature neurons are induced to undergo apoptosis after infection. In the more mature individuals, virus infection of neurons may not lead to apoptosis and in the absence of an effective immune response persistent infection is established. The major mechanism by which alphavirus infection of neurons is controlled is by production and local secretion of antibody to the surface glycoproteins. Antibody acts synergistically with interferon to decrease intracellular virus replication but does not eliminate the infected cell or the viral RNA within the cell. Therefore, the immune response controls, but does not cure the infection, leading to a requirement for long-term local synthesis of antiviral antibody in the CNS. For the host persistent intracellular RNA and need for longterm control is probably a reasonable price to pay for a noncytolytic mechanism for control of neuronal viral infection. PMID- 8725564 TI - The role of prostaglandins in gastric mucosal protection. PMID- 8725565 TI - Winner of the Theodore E. Woodward Award: c-Myb and the coordinate regulation of thymic genes. PMID- 8725567 TI - At risk nephrons and the decline in renal function in response to treatment of hypertension. AB - An examination of change in renal function following blood pressure lowering in more than 4,400 individuals in several clinical trials revealed that renal function declined following initiation of antihypertensive treatment in both essential hypertension and hypertensive diabetics for a period of two years before stabilizing at or near zero change. This initial decline can be related to the severity of preexisting hypertension but does not appear related to the type of antihypertensive regimen used. This phenomenon appears most readily explained by progressive obsolescence of previously damaged nephrons and not by the type of antihypertensive therapy employed. These finding raise questions about validity of interpretation of clinical trials designed to test efficacy of specific drug regimens in preserving renal function when outcome results are predominantly influenced by events during the first two years of intervention. PMID- 8725566 TI - Lipoprotein(a): searching for a function. PMID- 8725568 TI - New challenges to the therapy of systemic fungal infections. AB - In summary, the recent increase in frequency of systemic fungal infections has stimulated the development of new antifungal agents which are easier to use and which have decreased toxicity. This has resulted in increase in use, and along with this, the appearance of fungi resistant to antifungal agents. The medical community will have to come to terms with this newly emerging problem. PMID- 8725569 TI - Contrasting experiences and perceptions of women and men physician graduates of Stanford University School of Medicine. PMID- 8725570 TI - The effects of race and body fat distribution on insulin sensitivity. PMID- 8725571 TI - The Jeremiah Metzger Lecture on the climate of cities. PMID- 8725572 TI - Diabetes in minorities: reflections on the medical dilemma and the healthcare crisis. AB - In summary, diabetes in minorities is clearly on the increase. The complication rates are likewise increasing, contributing to the large burden of health care costs. This is a significant medical dilemma inasmuch as these high-risk populations are among the fastest-growing segments of our society. Fortunately, delay and prevention of complications are now possible, although greater programmatic interventions will be required. The absence of such interventions is a reflection of the dimensions of our health care crisis. What is more, we have such poor insight into the reasons for the higher risks and rates of disease and its complications. This means that a critical step toward resolution of this dilemma is more research. PMID- 8725573 TI - Cultural competence--an essential hybrid for delivering high quality care in the 1990's and beyond. AB - Establishing guidelines for culturally competent medical care will help all physicians to fulfill their mandate to meet the health care needs of the individual patient as well as of patient populations, and to lower costs by encouraging a healing partnership with the patient, thus increasing patient responsibility for his or her own health. Being culturally sensitive is not enough. Nor is simply classifying patients according to race adequate. Many researchers in social science and health services increasingly agree that race in our heterogeneous U.S. population has limited biological meaning and more often than not is just a poor proxy for culture or socioeconomic status. Guidelines and quality indicators that seek to measure and improve cultural competence must take into account, in an integrated fashion, these three necessary components in the delivery of high-quality services to populations: 1) the health-related cultural factors; 2) the incidence and prevalence of diseases in the population; and 3) treatment outcomes peculiar to that population. To be culturally competent is to incorporate and integrate these critical factors into caring for diverse populations. If health care systems, individually and collectively, are to provide care of high quality that is cost-effective to all populations, researchers and funders of research must invest in an aggressive agenda that pursues two directions: the validation of existing quality indicators in minority populations, and the development of new quality indicators that assess the organization's ability to develop culturally competent care. Only in this way will those of us in the medical profession be able to fulfill our calling to relieve suffering without discriminating against some populations. PMID- 8725574 TI - Funding for the colleges of medicine: integrated delivery systems to the rescue. PMID- 8725575 TI - Osler meets the marketplace--speculations on the future of internal medicine in the 21st century. PMID- 8725576 TI - Ranking hospitals by the quality of care for medical conditions: the role of complications. PMID- 8725577 TI - Morning report and work rounds: opportunities for teaching and learning. PMID- 8725578 TI - Clinical challenges in the climate of prison. PMID- 8725579 TI - Precancerous and cancerous lesions of the oral cavity. AB - The purpose of this article is to provide the clinician with the clinicopathologic characteristics of oral cancer and its precursors. As can be seen by the previous discussion, numerous variables influence the formation of these lesions, and as yet we are unable to predict the progression of any single, premalignant lesion with certainty. Because of this, only astute clinicians, armed with a healthy level of suspicion, will be able to affect the prognosis of oral malignancies by recognizing these conditions at the earliest possible stage. PMID- 8725580 TI - Viral infections in the immunocompetent patient. AB - Human herpesviruses and human papillomaviruses are ubiquitous and may cause an array of oral disease. Herpesviruses are unique because recurrent localized disease occurs after the primary infection. Human papillomaviruses are known for their ability to produce benign epithelial proliferations and increasingly are implicated in the development of oral cancer. This article reviews the pathogenesis of common oral lesions induced by these viruses. PMID- 8725581 TI - Recurrent aphthous stomatitis. AB - Recurrent aphthous stomatitis (RAS) is the most common oral mucosal disease in North America. In some instances, RAS represents the central feature of the multisystem disease complex Behcet's syndrome. This article reviews the clinical features, contributing etiologic factors, and etiopathogenesis of RAS and Behcet's syndrome and describes therapeutic considerations and strategies essential to management of patients suffering from recurrent mouth ulcers. PMID- 8725582 TI - Conditions peculiar to the tongue. AB - The tongue is the organ of taste, and it functions in speech, mastication, and the deglutition of food. This muscular organ is affected by many conditions and diseases, including a variety of developmental, traumatic, inflammatory, infectious, and neoplastic lesions. PMID- 8725583 TI - Clinical management of oral and perioral candidosis. AB - Oral candidosis is a diagnosable and treatable mucosal disease that may be complicated by various other local or systemic disease conditions. Dependable diagnosis always should include a combination of clinical signs and symptoms suggestive of candidal disease as well as positive cytologic or direct culture results. The management of candidal infections should be individualized for each patient, with concern for interactions with current medications, immune status, other concurrent mucosal diseases, and exogenous infectious sources. In selecting the appropriate therapeutic agent(s) the clinician should consider patient health factors, location and severity of infection, and the probability of chronicity. PMID- 8725584 TI - Diagnosis and management of oral lichen planus. AB - OLP has many clinical presentations. Some lesions of OLP require no treatment. Some must be managed for 20 years or more. The differential diagnosis of OLP is difficult and varied. Dermatologists should consider consultation with a dentist or specialist in oral medicine for diagnosis and management of lesions of OLP. PMID- 8725585 TI - Vesiculobullous disease of the oral cavity. AB - This article reviews the clinical features of vesiculobullous diseases with oral manifestations. Oral vesiculobullous diseases may be viral, immunologically mediated, drug induced, hereditary, or traumatic in origin. A general approach to the diagnosis is outlined, including immunofluorescent testing of suspected vesiculobullous disease. Recommendations for treatment of oral vesiculobullous diseases are included. PMID- 8725586 TI - Oral manifestations of selected sexually related conditions. AB - The incidence of certain microbial infections occurring in the oropharynx is well documented, but infection by other pathogens, although highly probable, has not been established with certainty. Considerable recent interest in the incubation period of various infections and improved community surveillance programs combine to ascribe infection to specific incidents. It is prudent for health care personnel to acknowledge the presence of many sexually transmitted diseases in the oropharynx and consider them in the differential diagnosis of many well established conditions. PMID- 8725587 TI - HIV-associated lesions. AB - Oral lesions are common in HIV infection and may be the first sign of AIDS. This article reviews the oral fungal and viral infections commonly detected in HIV infected patients, particularly candidiasis, deep fungal infections, herpes simplex virus infections, cytomegalovirus infections, and oral hairy leukoplakia. The neoplasms associated with AIDS such as oral Kaposi's sarcoma and lymphoma are related periodontal diseases. Each disorder is discussed by clinical appearance, diagnosis, and management. Recent advances in therapy are stressed. PMID- 8725588 TI - Lesions associated with drug reactions. AB - Several of the more common oral lesions secondary to medication use have been presented in this article. Although some of these conditions are specific to the oral tissues, some are not. Numerous cutaneous reactions to medications have mucosal counterparts. It is important for the clinician to recognize that many oral lesions are simply a manifestation of a systemic condition and should be managed with that in mind. PMID- 8725589 TI - Burning mouth syndrome. AB - Burning mouth syndrome is a common condition particularly affecting elderly women. Numerous precipitating factors are recognized that lead to a burning sensation in clinically normal mucosa. By taking each precipitating factor into account, a favorable treatment outcome usually can be achieved. This article highlights the significance of precipitating factors in burning mouth syndrome and suggests a treatment protocol based on current scientific evidence. PMID- 8725590 TI - Swellings of the oral cavity. AB - Many lesions that appear in the oral cavity manifest as a nodule or swelling. A particular lesion may have the clinical characteristics of several entities, and a differential diagnosis should be generated, with a final diagnosis reached by microscopic examination. This article discusses and shows the clinical features of some of the more common oral lesions or conditions that present as localized or generalized swellings, and it includes diagnoses and suggestions for management. PMID- 8725591 TI - Melkersson-Rosenthal syndrome and orofacial granulomatosis. AB - The Melkersson-Rosenthal syndrome is a rare disorder of unknown etiology characterized by a triad of recurrent orofacial swelling, relapsing facial paralysis, and fissured tongue. Exacerbations and recurrences are common. The orofacial swelling is characterized by fissured, reddish-brown, swollen, nonpruritic lips or firm edema of the face. The facial palsy is indistinguishable from Bell's palsy. The fissured tongue is seen in one third to one half of patients and, although the least common manifestation, its presence assists in diagnosis. The classic triad is not seen frequently in its complete form; therefore, diagnosis is difficult. This is particularly true because monosymptomatic and oligosymptomatic variants are seen more commonly. Cheilitis granulomatosa of Miescher is an example of a monosymptomatic variant of the Melkersson-Rosenthal syndrome. The histologic findings of noncaseating, sarcoidal granulomas support the diagnosis. These granulomas are not invariably present, and their absence does not exclude the diagnosis of the Melkersson-Rosenthal syndrome. Thus, the Melkersson-Rosenthal syndrome is a disease with elements of orofacial granulomatosis. Orofacial granulomatosis is a clinicopathologic entity describing oral lesions with noncaseating granulomas. The spectrum of this entity includes patients with oral Crohn's disease, patients with oral lesions who will develop typical bowel symptoms of Crohn's disease in the ensuing months to years, patients with tooth-associated infections, patients with sarcoidosis, and patients with food or contact allergies. The value of the clinicopathologic construct of orofacial granulomatosis is to provoke the careful search for provocative causes for the reactive symptom complex of the Melkersson-Rosenthal syndrome. PMID- 8725592 TI - Management of patients with epidermolysis bullosa. AB - Epidermolysis bullosa is a group of systemic disorders whose management requires familiarity with its many extracutaneous complications. These include gastrointestinal, ophthalmologic, laryngeal, dental, and hematologic problems. This article reviews wound care and management of systemic complications seen in patients with epidermolysis bullosa. PMID- 8725593 TI - [Effects of propofol on contractility and electrophysiological properties of canine single cardiomyocytes]. AB - We assessed the effects of propofol on contraction, action potentials and Ca2+ currents (Ica) in single ventricular cells in dog. Low concentrations of propofol 10(-5)M had no effects on contractile responses, but high doses above 10(-4)M of propofol decreased contractions. Propofol 10(-3)M reduced the duration of action potentials (time to 90% repolarization). Ica was decreased by propofol 10(-4)M without accompanying use-dependent manner. The present study indicates that clinical concentrations of propofol do not exert direct negative inotropic effects, but at supratherapeutic concentrations it inhibits cardiac contractility. PMID- 8725594 TI - [Isoflurane and sevoflurane impair left ventricular relaxation in dogs with fixed heart rate]. AB - The effects of isoflurane (Iso) and sevoflurane (Sev) on left ventricular relaxation were evaluated in 22 open-chest dogs with fixed heart rate (130 beats.min-1) using atrial pacing. Fentanyl was injected intravenously to maintain anesthesia during the preliminary preparation. In both Iso and Sev groups (n = 11), left ventricular systolic pressure, mean aortic pressure and dp/dt max were significantly decreased at 0.5 MAC, but there was no significant change in left ventricular end-diastolic pressure. Left ventricular systolic function was depressed to the similar extent in both groups. In Sev group, -dp/dt max and time constant of isovolumic left ventricular pressure fall (T) increased significantly at 0.5 MAC but it increased at 1.5 MAC in Iso group. T at 0.5 MAC Sev was also significantly longer compared with T at equipotent Iso. These findings suggest that Sev may impair isovolumic left ventricular relaxation more strongly than Iso, and this may result from the difference of the effect of each agent on intracellular Ca2+ homeostasis in the myocardium. PMID- 8725595 TI - [Subarachnoid administration of lidocaine reduces delayed neuronal damage due to forebrain ischemia in rats]. AB - We examined the effect of subarachnoid administration of lidocaine on delayed neuronal damage due to forebrain ischemia in rats. Sixteen rats were divided at random into two groups: lidocaine group rats (n = 8) and control group rats (n = 8) were subarachnoidally administrated lidocaine 5mg.kg-1 and normal saline, respectively, prior to ischemia. Forebrain ischemia was induced by bilateral carotid artery occlusion combined with systemic hypotension for 10 min. Cerebral perfusion-fixation was performed 7 days after ischemia, subsequent to which the brains were sectioned coronally and stained with celestine blue/acid fuchsin. In the neocortex and the hippocampal CA4 region, the lidocaine group showed less ischemic neuronal injuries than the control group. However, in the hippocampal CA1/CA3 regions and the caudoputamen, neuronal injuries in the lidocaine group were not significantly different from those in the control group. We conclude that the pre-ischemic subarachnoid administration of lidocaine reduces the delayed neuronal damage in neocortex and hippocampal CA4 region in the rat with forebrain ischemia. PMID- 8725596 TI - [Epidural buprenorphine does not improve diaphragmatic function after upper abdominal surgery]. AB - We investigated the effect of epidural buprenorphine on diaphragmatic function using respiratory inductive plethysmography (RIP) in seven healthy patients after upper abdominal surgery. After surgery, changes of rib cage contribution to tidal volume (%RC) increased significantly from 25.3 +/- 7.3 (mean +/- SD) to 50.7 +/- 14.8% (P < 0.05). After the injection of epidural buprenorphine, visual analogue scale (VAS) score decreased significantly from 8.3 to 3.3 (P < 0.05). But, %RC was unchanged compared to the value before the injection. These results indicate that pain relief by epidural buprenorphine does not improve diaphragmatic function after upper abdominal surgery. PMID- 8725597 TI - [Linear polarized light irradiation around the stellate ganglion area increases skin temperature and blood flow]. AB - We evaluated the effect of linear polarized light irradiation with a SUPER LIZER around the stellate ganglion area on skin temperature and blood flow in healthy adult volunteers. We carried out two experiments (study I and study II). In study I, we investigated one-sided irradiation around the stellate ganglion area or posterior neck on the skin temperature of the bilateral nasus externi and earlobes. In study II, we investigated one-sided irradiation around the stellate ganglion area or posterior neck on the skin temperatures of both hands and skin blood flow on the irradiated side. In study I, irradiation around the stellate ganglion area increased skin temperature on the irradiated sides of the nasus externi (wings of the nose), and in study II, skin temperature and blood flow were increased on the irradiated side of the hands. These results suggest that linear polarized light irradiation with a SUPER LIZER around the stellate ganglion area would be useful and beneficial in clinical therapy. PMID- 8725598 TI - [Concurrent characterization of spinal amino acid release and touch-evoked allodynia produced by spinal glycine or GABAA receptor antagonists]. AB - BACKGROUND: Previous work has shown that spinal strychinine (STR; glycine antagonist) or bicuculline (BIC; GABAA antagonist) yields a touch-evoked allodynia (TEA) that was blocked by NMDA receptor antagonists and that spinal NMDA receptor activation evokes glutamate release. In the present study, we examined the effects of spinal STR and BIC on TEA and the spinal release of amino acids, and spinal NMDA receptor activation with intrathecal NMDA. METHODS: In halothane-anesthetized (1.0%) Sprague-Dawley rats, TEA was assessed after intrathecal injection of STR (3 micrograms) or BIC (10 micrograms). TEA was scored as 0: no appearance to 3: strong agitation and was evaluated at 10-min intervals after injection of the drugs. Simultaneously, spinal release was monitored by using a microdialysis probe implanted into the lumbar subarachnoid space four days prior to the experiment. Glutamate (Glu) and taurine (Tau) in dialysates were measured by HPLC-UV. To assess the contribution of presynaptic glutaminergic neuron, the effects of intrathecal NMDA (3 micrograms) combined with either STR or BIC on spinal release were also examined. RESULTS: Time course of average TEA scores after intrathecal STR or BIC was similar at 10, 20 and 30 min (STR: 1.6, 0.3, 0; and, BIC: 1.8, 0.9, 0.1). Intrathecal BIC alone evoked a transient spinal release of Glu and Tau with increases by 54% and 68% respectively in the 0-10min samples while STR exerted no effect. Intrathecal NMDA faciliated the effects of BIC and STR on TEA score and spinal release of Glu and Tau. CONCLUSION: It is suggested that the development of allodynia may result from either a loss of GABAA inhibition on the presynaptic terminals and/or by a loss of postsynaptic glycine inhibition, the former facilitating spinal glutamate release, and the latter not. PMID- 8725599 TI - [The effects of preoperative drinking and H2 blocker on gastric acid secretion]. AB - We studied the effects of preoperative drinking and H2 blocker on gastric acid secretion in 63 patients (ASA I-II, > 18yrs) scheduled for afternoon surgery. Group A (n = 20), as a control, was not permitted to eat and drink from 9 pm, the day before surgery, and was then given 500 ml of maintainance fluid before anesthesia. Group B (n = 20) fasted from 9 pm the day before surgery, and was allowed to drink clear fluids until 2hs before anesthesia. Group C (n - 23) followed the same guidelines as group B, and was given famotidine (20mg) orally at 9 pm the day before and 2hs before anesthesia. After induction, a Salem sump tube was inserted into the stomach and a gastric fluid aspiration was performed. The fluid volume and pH were measured after collection. Gastric pH was significantly higher (P < 0.001) in group C (6.4 +/- 0.9) than in groups A (3.1 +/- 1.8) and B (2.7 +/- 1.8). Fluid volume was similar in each group (A; 11 +/- 9/B; 12 +/- 9/C; 12 +/- 13ml). The dilution of gastric acid by the ingested fluid was not observed. We conclude that preoperative drinking does not affect gastric contents in elective operative patients. To reduce the risk of developing aspiration pneumonia, we recommend that every patient should receive an oral H2 blocker. PMID- 8725600 TI - [Intraoperative acute myocardial infarction during total gastrectomy under general anesthesia combined with thoracic epidural anesthesia]. AB - A 61-year-old, 54-kg man with gastric cancer was scheduled for total gastrectomy under general anesthesia combined with thoracic epidural anesthesia. Approximately 20 minutes after the start of the operation, the patient developed sudden hypotension and ventricular fibrillation. The cardiac rhythm returned to normal after 38 minutes of cardiopulmonary resuscitation. The operation was discontinued and the patient was transferred to an intensive care unit. A 12-lead electrocardiogram (ECG) revealed complete right bundle branch block and elevation of the ST-segment from leads II, III, aVF, V1, and V2 which indicated an inferior myocardial infarction. Laboratory data showed elevated levels of enzymes such as glutamic oxaloacetic transaminase (495 IU.l-1), lactic dehydrogenase (1781 IU.l 1), and creatine kinase-MB (112 IU.l-1). The mildly elevated levels of the enzymes decreased around 10 hours after the termination of the operation, but they increased markedly again without any change in ECG on the second postoperative day. Elevation in serum myoglobin and glutamic pyruvic transaminase and decline in arterial ketone body ratio were also detected. Furthermore, renal failure developed with increase in blood urea nitrogen and creatinine. Because of hepatic failure and renal failure which might have been caused by rhabdomyolysis, the patient needed inotropic support with dopamine, dobutamine, and epinephrine to maintain the circulation. The patient died of reinfarction of the 20th postoperative day despite intensive care such as intraaortic balloon pumping, hemodiafiltration, and continuous intravenous infusion of prostaglandin E1. PMID- 8725601 TI - [Permanent paraplegia following epidural and general anesthesia: two case reports]. AB - Case-1: A 72-year-old woman with no past neurological history was scheduled for a rectum resection under general combined with epidural anesthesia. An epidural catheter was introduced at T11-12 interspace without any difficulties. During the operation, she had hypotensive episode needing dopamine, but waked up from anesthesia without any event. When she became alert, she complained muscle weakness and loss of sensation in both lower extremities. On the day after surgery, she became quadriplegic and completely insensitive under Th4 level, but her MRI of the spine showed no abnormal findings. A month after the operation, her MRI showed diffuse spinal degeneration below C4 level and she had flaccid paralysis below Th1 with complete sensory loss below Th7 level. Case-2: A 62-year old man with no past neurological history was scheduled for gastrectomy under general combined with epidural anesthesia. An epidural catheter was placed via T12-L1 without any difficulty. Operative course was uneventful and awakening from anesthesia was normal. He showed muscle weakness and hypesthesia of lower extremities two hours after the operation, and we stopped continuous injection of epidural anesthesia. His paralysis became worse but MRI of his spine showed no abnormality on the day after the operation. He became complete flaccid paralytic and had complete sensory loss below T7 level. The MRI examination two weeks after the operation showed degeneration below middle thoracic spinal cord. His neurologic symptoms have not improved for two years. The etiology of neurologic deficits of these two case is not obvious although the relation between epidural anesthesia and neurologic symptoms was most likely. PMID- 8725602 TI - [Perioperative managements of a mentally retarded child with obstructive sleep apnea syndrome for adeno-tonsillectomy]. AB - We describe a mentally retarded child with obstructive sleep apnea syndrome (OSAS) in whom it was difficult to maintain upper airway in the perioperative period. The child underwent awake intubation, because the preanesthetic evaluation of the airway with a direct fiberoptic visualization revealed a very narrow airway. Also we considered that if we used an anesthetic agent, a perioperative airway management would be very troublesome. Postoperatively we continuously monitored for apnea and arrhythmias. When the child was sleeping, we found frequently that her thoratic movements were getting weak and percutaneous oxygen saturation went down to about 70 percent. The preoperative direct fiberoptic visualization of the upper airway is effective for the evaluation of the degree of airway obstruction in this child. We also recommend the continuous intensive postoperative monitorings including pulse oximetry, ECG, and apnea monitor which are very important to avoid life-threatening complications such as upper airway obstruction and serious arrhythmias in patients with obstructive sleep apnea syndrome. PMID- 8725603 TI - [General anesthesia for expandable endotracheobronchial metal stent insertion]. AB - We experienced eight cases of general anesthesia for endotracheobronchial expandable metal stent insertion. Awake intubation was performed following NLA induction. Anesthesia was maintained by inhalational anesthetics and fentanyl was added under spontaneous breathing assisted manually. Two patients showed a marked hemodynamic change and hypoxia due to endotracheobronchial bleeding and suction. Six patients received EMS insertion with no trouble. We conclude that endotracheobronchial bleeding is an important factor to be watched for during anesthetic management for EMS insertion. PMID- 8725604 TI - [Hemilateral hydrothorax and atelectasis during laparoscopic Nissen fundoplication]. AB - A 57-year-old woman, weighing 75 kg, with gastroesophageal sliding hernia, received laparoscopic Nissen fundoplication under general anesthesia. Although artificial ventilation was carried out uneventfully when the surgery started, sudden decreases in SpO2 and thoraco-pulmonary compliance were noted after insufflation of CO2. Breath sound was not audible on the left lung. We suspected that inadvertent pneumothorax was produced, but chest X-ray and transesophageal echocardiography at the end of surgery revealed the presence of left hemilateral hydrothorax with pulmonary atelectasis. She was ventilated artificially and given diuretics and albumin solution for 3-days before extubation. We speculated on two reasons for the event: a possibility of perforation of the diaphragm manipulated by surgeons, and that of transition of fluid used for irrigation just below the diaphragm. Pulmonary atelectasis could be induced with hemilateral lung ventilation because cephalad shifting of the diaphragm might follow the intraperitoneal insufflation. We conclude that laparoscopic surgery with insufflation of gas and irrigation with fluid requires careful attention if the laparoscopic surgery is sustained with insufflation and irrigation. PMID- 8725605 TI - [Postoperative nausea, vomiting and pain in laparoscopic cholecystectomy: a comparison with minilaparotomy-cholecystectomy]. AB - Postoperative nausea, vomiting and pain were compared between laparoscopic cholecystectomy group and minilaparotomy-cholecystectomy group. All patients were women, and ranged in age from 20 to 60 years. The body mass index of the patients was less than 30, and duration of operation was within 120 minutes in both groups. All patients received general anesthesia combined with epidural analgesia. Morphine hydrochloride 4 mg was administered into epidural space before incision. No significant differences were found in the incident of nausea and vomiting among the two groups. Postoperative analgesic requirement of laparoscopic cholecystectomy group was significantly less than that of minilaparotomy-cholecystectomy group. No significant differences were found in the incidence of nausea and vomiting between the group which required postoperative analgesic drugs and the group which required no postoperative analgesic drugs. These results suggest that laparoscopic operation and postoperative pain do not influence the incidence of nausea and vomiting. Postoperative pain after laparoscopic cholecystectomy is less than that after minilaparotomy-cholecystectomy. PMID- 8725606 TI - [Accuracy of a pulse oximeter during hypoxia]. AB - The accuracy of the pulse oximeter was examined in hypoxic patients. We studied 11 cyanotic congenital heart disease patients during surgery, and compared the arterial oxygen saturation determined by both the simultaneous blood gas analysis (CIBA-CORNING 288 BLOOD GAS SYSTEM, SaO2) and by the pulse oximeter (DATEX SATELITE, with finger probe, SpO2). Ninty sets of data on SpO2 and SaO2 were obtained. The bias (SpO2-SaO2) was 1.7 +/- 6.9 (mean +/- SD) %. In cyanotic congenital heart disease patients, SpO2 values were significantly higher than SaO2. Although the reason is unknown, in constantly hypoxic patients, SpO2 values are possibly over-estimated. In particular, pulse oximetry at low levels of saturation (SaO2 below 80%) was not as accurate as at a higher saturation level (SaO2 over 80%). There was a positive correlation between SpO2 and SaO2 (linear regression analysis yields the equation y = 0.68x + 26.0, r = 0.93). In conclusion, the pulse oximeter is useful to monitor oxygen saturation in constantly hypoxic patients, but the values thus obtained should be compared with the values measured directly when hypoxemia is severe. PMID- 8725607 TI - [Transient complete left bundle branch block during epidural anesthesia with mepivacaine]. AB - A 65-year-old woman with ovarian tumor was scheduled for abdominal total hysterectomy and bilateral adnexectomy under epidural anesthesia. Preoperative examinations revealed no abnormalities including ECG. Twenty minites after the epidural injection of mepivacaine, widened QRS complexes, which were diagnosed as complete left bundle branch block (CLBBB) later, appeared on ECG. At that time, heart rate (HR) was 150.min-1. CLBBB disappeared when HR decreased to 110.min-1 following the administration of pindolol and verapamil. The operation was postponed. Exercise-treated ECG showed neither CLBBB nor ischemic change even when HR increased to 160.min-1. Two weeks later the operation was attempted again. CLBBB appeared again twenty minites after the epidural injection of mepivacaine. The CLBBB disappeared 5 minites after the induction of general anesthesia when HR decreased to 100.min-1. The operation was performed uneventfully. The effective refractory period (ERP) is shortened with the increase in HR, but the shortning of ERP varies in different part of the cardiac conduction system. In tachycardia, ERP of left bundle branch is longer than that of right one. Because the cardiac conduction system is depressed by local anesthetics, it is speculated that ERP of left bundle branch is elongated further by mepivacaine and CLBBB appeared in this case. PMID- 8725608 TI - [Usefullness of transesophageal echocardiography in early detection of coronary spasm]. AB - Intraoperative transesophageal echocardiography (TEE) was performed on a 62-year old man who underwent abdominal aortic replacement for abdominal aortic aneurysm under general anesthesia combined with epidural anesthesia. Coronary artery spasm occurred after unexpected massive hemorrhage, and TEE showed hypokinesis in the posterior-inferior left ventricular wall. The changes in TEE preceded the ST elevation in the ECG. Bolus infusion of isosorbide dinitrate and continuous infusion of nitroglycerin alleviated these changes. TEE enabled us to detect and evaluate coronary spasm before the appearance of ST changes in ECG. PMID- 8725609 TI - [Preanesthetic evaluation of cardiovascular reserve in a patient with dilated cardiomyopathy]. AB - We described an anesthetic management of a patient with abdominal aortic aneurysm associated with dilated cardiomyopathy (DCM) focusing on preanesthetic evaluation of cardiovascular reserve and on intraoperative continuous circulatory monitoring with transesophageal echocardiography (TEE) and continuous cardiac output measurement (CCO). Based on echocardiographic and hemodynamic measurements after a 50 m-walk load, we predicted the allowable range of alteration of preload (LV diastolic dimension; Dd), myocardial performance (arterial blood pressure and ejection fraction) and of heart rate. During anesthesia and operation, we continuously monitored Dd, arterial blood pressure, heart rate and cardiac output, and maintained these variables within the allowable range. The changes in preload after clamping or unclamping of the aorta was promptly reflected by Dd as compared to pulmonary capillary wadge pressure. The CCO was also usuful in detecting abrupt changes in myocardial performance. In conclusion, we suggest preanesthetic stress test to be performed to evaluate cardiovascular reserve and to predict the allowable range of alteration of hemodynamic variables. Continuous monitoring of preload (Dd) by TEE and of myocardial performance by CCO is useful to detect early changes in these variables. PMID- 8725610 TI - [Acute alcoholism after ethanol fixation for ovarian chocolate cyst]. AB - We experienced a case of acute alcoholism after ethanol fixation for ovarian chocolate cyst. A 46 year old female was scheduled for ethanol fixation for ovarian chocolate cyst. Ethanol fixation was performed with 110 ml of 99% ethanol under general anesthesia. Ethanol 70 ml was not removed. After the operation, patient did not emerge from anesthesia within 30 min. Blood alcohol concentration was 232 mg.dl-1. It is important to remove total ethanol used for ethanol fixation in anesthetic management for ethanol fixation for ovarian chocolate cyst. PMID- 8725611 TI - [Dextran-induced anaphylactoid reactions in two patients with gastrointestinal cancer]. AB - We report dextran-induced anaphylactoid reactions (DIAR) subsequent to rapid infusion of Rheomacrodex (dextran 40) in two patients, a 67 year old man with gastric cancer undergoing distal gastrectomy and a 47 year old man with transverse colon cancer undergoing colectomy. Both showed sudden tachycardia, hypotension and skin flush, which were treated with epinephrine or etilephrine administration. Most cases of severe DIAR are immune complex anaphylaxis mediated by dextran-reactive antibodies (DRA) of the IgG class, which are considered to arise mainly in response to immunization with dextran-cross-reactive bacterial polysaccharides in the gastrointestinal tract. High titers of DRA have previously been reported in gastric ulcer patients with pyloric stenosis, suggesting bacterial polysaccharides permeation through the luminal wall which may easily occur in the presence of local inflammation or ulcer. Although serum DRA titers in our patients have not been examined, inflammation or ulcer around the tumor might have played a role in producing high titers of DRA. In patients suspected of gastrointestinal ulcer or inflammation, including cancer, dextran administration is not preferable or should be avoided, unless hapten-dextran preparation is used for the prophylaxis of severe DIAR. PMID- 8725612 TI - [Has the lifesaving technician system improved the outcome of out-of-hospital cardiopulmonary arrest patients?]. AB - The role of the lifesaving technician (LST) system on the outcome of out-of hospital cardiopulmonary arrest patients was examined retrospectively. The percentage of the patients resuming spontaneous circulation before arrival at the hospital was 2.2-4.3 times higher in the group with the prehospital care by LSTs ('93: 3.9% and '94: 2.0%) than by ordinary ambulance personnel (OAP) (0.9%), but there was no difference in the percentage of return of spontaneous circulation (LST '93: 33.0%, '94: 27.3%, OAP 37.8%), survival rate (LST '93: 3.1%, '94: 1.4%, OAP 1.8%) and discharge rate (LST '93: 1.0%, '94: 0.7%, OAP 1.8%) of the patients with cardiopulmonary arrest on arrival. Time interval from fire department receipt of call to ambulance departure and arrival at the hospital was significantly longer in the groups with the prehospital care by LSTs than by OAP. There was no difference in the outcome of cardiopulmonary arrest patients with the prehospital care by LSTs between '93 and '94. Our results suggest that making much more effort with more advanced technics to start the spontaneous circulation on the spot may be required to improve the outcome of out-of-hospital cardiopulmonary arrest patients. PMID- 8725613 TI - The twenty years of Hans Gurland. PMID- 8725614 TI - Membranes in artificial organs. AB - Membrane processes by a pivotal and enabling role in modern replacement therapy for acute and chronic organ failure and in the management of immunologic diseases. In fact, virtually all contemporary extra-corporeal blood purification methods employ membrane devices, and the next generation of artificial organs and tissue engineering therapies are almost certain to be similarly grounded in membrane technology. In this short essay, we comment on the similarities and differences among synthetic membranes and their natural counterparts and also provide a critical overview of the demographics and technology of hemodialysis, hemofiltration, apheresis, oxygenation, and emerging membrane technologies and applications. PMID- 8725615 TI - Characterization of molecular transport in artificial kidneys. AB - Transport characterizations of artificial kidneys require the use of multiple marker molecules of various sizes, including small solutes, middle molecules, and albumin. Previous approaches for evaluating hemodialyzer transport performance are reviewed. New data obtained from in vitro experiments comparing 5 low molecular weight proteins of approximately the same molecular size as markers for middle molecule transport also are described. Sieving coefficients for marker low molecular weight proteins may vary substantially for a given artificial kidney membrane. Furthermore, sieving coefficients for marker proteins that do not absorb significantly to the membrane are comparable to those for polydisperse dextrans. These observations suggest that other protein properties besides molecular size are important determinants of protein sieving coefficients across artificial kidney membranes. We conclude that low molecular weight proteins can behave differently from one another and generalizations about artificial kidney membrane transport from data obtained on a single protein may be problematic, and that both low molecular weight proteins and polydisperse dextrans are useful markers of middle molecular transport across artificial kidney membranes. PMID- 8725616 TI - Soluble TNF alpha receptors are increased in chronic renal insufficiency and hemodialysis and inhibit neutrophil priming by TNF alpha. AB - The oxidative burst of neutrophils from azotemic patients is refractory to priming by tumor necrosis factor-alpha (TNF alpha). Soluble TNF alpha binding protiens (sTNFR) accumulate in the plasma of azotemic patients. To test the hypothesis that these increased sTNFR concentrations inhibit TNF alpha priming of oxidative burst activity, we measured plasma sTNFR concentrations in nondialyzed azotemic patients, hemodialysis patients, and normal subjects, and determined TNF alpha priming of fMet-Leu-Phe-stimulated superoxide production in neutrophils incubated in plasma with differing levels of sT-NFR. These sTNFR concentrations increased significantly as creatinine clearance decreased and were significantly greater in hemodialysis patients than could be accounted for by loss of renal function alone. TNF alpha primed superoxide production by normal neutrophils in normal plasma, but this effect was significantly reduced in plasma with increased concentrations of sTNFR. Neutrophils from azotemic and hemodialysis patients were refractory to priming by TNF alpha in autologous plasma, and incubation in normal plasma only partially corrected this defect. We conclude that sTNFR accumulate as a result of the loss of renal function and hemodialysis and inhibit TNF alpha priming of neutrophils in azotemic and hemodialysis patients, but that these cells also have an intrinsic functional defect. PMID- 8725617 TI - Ex vivo manipulation of cell subsets for cell therapies. AB - Large-scale cell separation and ex vivo expansion technologies will form the basis for development of new cellular products for the treatment of cancer and fatal viral diseases. The cell subsets that are likely to play a significant role in cellular therapy include hematopoietic stem cells, platelet and granulocyte precursors, cytotoxic lymphocytes, and genetically modified hematopoietic or lymphoid precursors. Cell enrichment techniques are required to eliminate tumor cells from autologous stem cell grafts and to reduce the size of culture systems required for expansion or gene transfection. The consumption of expensive culture components such as cytokines and serum may be reduced by the use of perfusion bioreactor devices. Methods that have been developed for the production of cell subsets for cellular therapy are reviewed. PMID- 8725618 TI - Renal transplantation: progress and prospects. AB - Despite encouraging and improving results, organ transplantation is still hampered by a shortage of organs, chronic transplant loss, and a changed patient population. Liberal inclusion criteria for dialysis and/or renal transplantation and the increasing unwillingness to donate organs in some countries had led to a growing imbalance between the numbers of transplantations performed and patients on waiting lists. Until now, poorly understood chronic transplant dysfunction is responsible for a still unchanged graft loss of approximately 5% per year. The patient population has changed to include more multimorbidity and an increasing number of risk factors (age, diabetes mellitus, former [failed] transplantations, or preexisting cardiovascular diseases). The recommendation for a against dialysis or transplantation has become increasingly difficult for the responsible physician. Newly developed immunosuppressant drugs, an increasing consideration regarding living organ donation, or xenotransplantation in the future may solve this dilemma. New reflections and considerations about the ethical background of transplantation medicine are necessary. PMID- 8725620 TI - Lipid apheresis: from a heroic treatment to routine clinical practice. AB - Lipid apheresis has developed from a heroic treatment into a routine clinical therapy and currently is the major indication for performing extracorporeal plasma therapy. Whereas it was once reserved for patients with homozygous familial hypercholesterolemia, today it has a place in the secondary prevention of severe coronary heart disease when low-density lipoprotein (LDL)-cholesterol level exceeds 150 mg/dl, despite conservative treatment, in any type of primary hypercholesterolemia. Unselective plasma exchange has been replaced by a variety of selective procedures. The efficacy of the treatment can be maximized by combining LDL apheresis with the use of cholesterol synthesis enzyme inhibitors. Clinical studies have shown that drastic cholesterol reduction can result in regression of coronary atherosclerosis as well as in reduced cardiac morbidity and mortality. Technical progress comprises improved selectivity, online regeneration of adsorbers, and LDL adsorption from whole blood. Recently, a new LDL hemoperfusion procedure was successfully tested in a clinical pilot study; blood is passed directly over a lipid sorbent without prior plasma separation. If this system is demonstrated to be safe and effective in clinical Phase III trials, a further qualitative step in the rapid development of LDL apheresis will have made. PMID- 8725619 TI - Plasma therapy at Klinikum Grosshadern: a 15-year retrospective. AB - Immediately after the availability of highly permeable membranes in 1979, membrane plasma separation was introduced as a mode of extracorporeal blood purification by the nephrology group at Klinikum Grosshadern of the Ludwig Maximilians University of Munich (F.R.G.). The new therapy was applied primarily in the management of immunologically mediated renal and extrarenal disorders as well as in paraproteinemias. We also have witnessed a widespread application of this extracorporeal treatment as a last resort in otherwise refractory clinical conditions. Over the years, the group at Grosshadern has contributed to the development, as well as to the laboratory and clinical testing, of new plasma separation membranes, simplified plasmapheresis formats (e.g., spontaneous membrane plasma separation), and several plasma fractionation procedures (e.g., cascade filtration, adsorption). Whenever indicated and possible, plasma fractionation procedures, rather than unselective plasma exchange, are performed in an appropriate clinical situation. PMID- 8725621 TI - Microspheres based detoxification system: a new method in convective blood purification. AB - A variety of protein-bound or hydrophobic substances, accumulating as a result of pathologic conditions such as exogenous or endogenous intoxications, are removed poorly by conventional detoxification methods because of low accessibility (hemodialysis), insufficient adsorption capabilities (hemosorption), low efficiency (peritoneal dialysis), or economic limitations (high-volume plasmapheresis). Combining advantages of existing methods with microspheric technology, a module-based system was designed. Major operating parameters of the latter can be modified to allow for adjustment to individual clinical situations. An extracorporeal blood circuit including a plasmafilter is combined with a secondary high-velocity plasma circuit driven by a centrifugal pump. Different microspheric adsorbers can be combined in one circuit or applied in sequence. Thus, a prolonged treatment can be tailored using specially designed selective adsorber materials. Comparing this system with existing methods (high-flux hemodialysis, molecular adsorbent recycling system), results from our in vitro studies and animal experiments demonstrate the superior efficiency of substance removal. PMID- 8725622 TI - The significance of adhesion molecules in nephrology. AB - Our knowledge of adhesion molecules has exploded over the last 5 years and has swamped most fields of medicine including nephrology. This is not surprising because adhesion molecules play a pivotal role in all aspects of cell to cell contact. Thus, they are involved in important issues, such as fetal development, in any kind of inflammatory or immune response including allograft rejection, as well as thrombus formation, and in tumor growth and metastasis (1-3). This short overview briefly reports some aspects of the biology of relevant adhesion molecules and their significance in inflammatory kidney diseases and in hemodialysis and renal allograft rejection. Finally, new therapeutic opportunities that arise by blocking adhesion molecule function are discussed. PMID- 8725623 TI - Dilemma of membrane biocompatibility and reuse. AB - Numerous articles have been published on the multiple use of dialyzers and on the effect of different reprocessing chemicals and techniques on the dialyzer biocompatibility and performance. The results often appear contradictory, especially those comparing standard biocompatibility parameters. Despite this confusion, a discerning review of the published works allows certain limited conclusions to be drawn. Reprocessing of used hemodialyzers changes the biocompatibility profile of a dialyzer as defined by the parameters complement activation, leukopenia, and cytokine release. The effect of reprocessing depends on the chemicals and reprocessing technique applied and also on the type of membrane polymer being subjected to the reprocessing procedure. Reports of pyrogenic reactions indicate that the flux of the membrane also influences how suitable it is for safe reuse. An increased risk of allergic and pyrogenic reactions appears to be associated with dialyzer reuse. Furthermore, there has been a lack of investigations into the immunologic effect of the layer of adsorbed and chemically altered proteins that remains on the inner surface of reprocessed dialyzers. We conclude that the clinical benefit of dialyzer reuse cannot be generally accepted from a biocompatibility point of view. PMID- 8725624 TI - Molecular biology of cytokines. AB - The development of the technological armamentarium of molecular biology has revolutionized biomedical research in general and nephrologic investigation in particular. In addition to the recent identification of several genes involved in normal kidney function and pathologic conditions, our knowledge regarding the role of cytokines in primary renal diseases, transplant rejection, and dialysis effects has expanded greatly. In particular, molecular biologic methodology has provided insight into the mechanisms controlling cytokine gene regulation, which occurs primarily at the transcriptional level and is mediated by DNA-binding proteins interacting with specific recognition motifs in genetic promoter and enhancer elements. Interleukin-6 (IL-6) is discussed as an example because it is a secretory product of mesangial cells and participates in the cytokine network that determines glomerular and interstitial inflammation. In our analysis of IL-6 gene regulation employing reporter gene and electrophoretic mobility shift assays, we have found that bacterial lipopolysaccharide and cyclic adenosine monophosphate synergistically induce IL-6 expression in macrophages through at least four transcription factors, including AP-1, cAMP-responsive element-binding protein (CREB), NF-IL6, and NF-kappa B. One of the most exciting areas of future research will focus on transcription factor activation in experimental and clinical disease states. Novel therapeutic approaches targeting transcriptional regulation are currently being explored. PMID- 8725625 TI - Experimental glomerulosclerosis: Defektheilung of the kidney. AB - Research in the role of cytokines in experimental glomerulonephritis has increased our understanding of the mechanisms that may be involved in the development of progressive renal disease. Glomerulosclerosis, the final common pathway in a variety of underlying kidney diseases, is characterized by increased extracellular matrix formation and cell proliferation. Transforming growth factor beta (TGF-beta) and monocyte chemoattractant protein-1 (MCP-1) have been identified in animal models as mediators in the processes that follow renal injury. There is evidence of similar events occurring in other fibrotic disorders, suggesting that there is a common generic pathway of fibrosis. This review summarizes our knowledge of TGF-beta and MCP-1 in experimental kidney disease and compares these results with mechanisms described in other organs. We propose that glomerulosclerosis represents Defektheilung (healing by secondary intention) of the kidney after various injuries. The growing knowledge of the mechanisms involved will help advance future therapeutic interventions by directing the healing process toward primary healing. PMID- 8725626 TI - Hanta is coming. AB - The Hanta virus is coming again. Yes, it is coming again and again to your town to your rodents and to your homes! We can resolve to prevent it, contain it, and treat it in a scientific and humane way, or we can salivate, scratch, bite, and run like rats and mice and voles and shrews. PMID- 8725627 TI - Inconsequence of membrane choice in acute renal failure? AB - The choice of hemodialysis membrane in acute renal failure has caused a heated debate, principally because of the dogmatism with which the results of preliminary clinical studies have been translated into prescription dictum. The issue, however, is not merely the limitations of these two studies, but rather the shift in emphasis they may have engendered in the approach to dialytic therapy in acute renal failure. Dogmatism based on limited or flawed data does not serve the interests of our patients, and the issue of hemodialysis in acute renal failure is far more complex than the exaggerated importance of membrane choice. PMID- 8725628 TI - Developmental neurotoxicity of PCBs in humans: what do we know and where do we go from here? AB - The potential neurotoxicity of PCBs was first recognized in 1968 when a number of Japanese people became ill after ingesting rice oil that was contaminated with PCBs during the manufacturing process (Yusho). Later a similar exposure occurred in Taiwan (YuCheng). Children born to Taiwanese mothers who consumed PCB contaminated rice oil were followed and a number of developmental abnormalities, including lower body weight and height, higher activity levels, greater incidence of behavior problems, and lower IQ scores, were observed. However, interpretation of these findings is complicated by the fact that there did not appear to be any relationship between available indices of exposure and severity of effects, and by the fact that the PCBs to which the Taiwanese were exposed contained unusually high concentrations of dibenzofurans, which are many times more toxic than PCBs, and may have been responsible for some or all of the observed effects. Since the Yusho and YuCheng episodes, several studies have been initiated to study the neurobehavioral effects of exposure to the lower levels of PCBs present in the environment. The two studies published to date have yielded conflicting results. Jacobson, Jacobson, and colleagues reported that in utero PCB exposure was associated with decreased birth weight and head circumference, shorter gestation, and several adverse outcomes on the Brazelton Neonatal Assessment Battery. Later they reported that the body weight deficits associated with prenatal PCB exposure were still present at 5 months and 4 years of age. Deficits in memory function were observed at 7 months and 4 years. Rogan, Gladen, and colleagues did not find any evidence of decreased birth weight or head circumference. Nor did they find any evidence of deficits in memory function. However, they did observe some similar effects on the Brazelton Neonatal Assessment Battery. They also observed a small delay in psychomotor development in the most highly PCB-exposed children, but the effect did not persist beyond 2 years of age. A number of methodological concerns have been raised about the Jacobson study, including issues related to exposure assessment, sample selection, and control of potential confounding variables. However, it is not clear that these shortcomings can explain the discrepancies between their findings and those of Rogan and Gladen. Other possible explanations include differences in exposure levels or PCB congener patterns between the two cohorts, differences in sociodemographic variables between the two cohorts, or other problems inherent in trying to detect subtle neuropsychological deficits at exposure levels that are near the threshold for effects. Hopefully, several new studies that are currently underway will help to resolve the uncertainties regarding the risks of perinatal PCB exposure that have been created by the conflicting results of these early studies. PMID- 8725629 TI - PCBs and behavioral impairment: are there lessons we can learn from lead? PMID- 8725630 TI - Adopting a public health approach to developmental neurotoxicity. PMID- 8725631 TI - Taking a lead from wildlife. PMID- 8725632 TI - Epidemiologic perspective of the developmental neurotoxicity of PCBs in humans. PMID- 8725633 TI - Comments on "developmental neurotoxicity of PCBs in humans: what do we know and where do we go from here"? PMID- 8725634 TI - The need for cellular, biochemical, and mechanistic studies. AB - The results of diverse in vitro neurotoxicity studies demonstrate that there are variations in cellular responsiveness between different types of neural cells. In contrast to experimental systems that have reported cellular responses to relatively high concentrations of various PCB congeners, our studies with rat hippocampal neural cells indicate that neurons and astroglia are responsive to relatively low levels of TCDD. However, these responses are probably not mediated through the classical Ah receptor pathway, which involves nuclear Ah receptor mediated modulation of gene expression. It has recently been reported that TCDD induced phosphorylation and other responses can be observed in some cell lines within minutes after treatment (20), and that cell membrane or cytosolic receptors may also play a role in mediating these effects. Future studies are required to determine both Ah receptor-dependent and -independent pathways associated with the neurotoxicity of PCBs, TCDD, and related compounds. The report that low-level dietary or background exposure to HAHs (32) results in neurobehavioral deficits is still a perplexing problem also requiring additional research and consideration of other dietary factors that may contribute to these effects. For example, we have recently been comparing the toxic effects and relative potencies of TCDD (exodioxins) and other "natural occurring" compounds (endodioxins) such as indole-3-carbinol (vegetables) and polynuclear aromatic hydrocarbons (PAHs, cooked foods), which also bind to the Ah receptor. Dr. Clynn Wilker has shown that in utero exposure of rats to indole-3-carbinol and chrysene (a PAH) cause demasculinization of the adult offspring as previously reported for TCDD (19). Thus, the neurotoxicity of low-level dietary exposure to HAHs should at least consider other possible confounding factors, including dietary endodioxins. PMID- 8725635 TI - Can epidemiological studies discern subtle neurological effects due to perinatal exposure to PCBs? AB - What conclusions can be drawn concerning the potential neurological effects of perinatal exposure to either PCBs, or PCBs and other fish-borne contaminants? First, by their very nature epidemiological studies are limited in their ability to detect subtle associations--including possible links between exposure to low levels of environmental contaminants and disease. As stated by Dr. Schantz, both Rogan and the Jacobsons report small changes in motor and cognitive behavior- typically less than one-half of a standard deviation--and only in the most highly exposed children. Given these small changes in CNS function, the substantive criticisms of Paneth (including the Jacobsons' choice to employ a random, rather than matched, control sample and the related fact that fish-eating mothers differed from non-fish-eating mothers on several important characteristics) and similar "generic" concerns raised by Taubes, a critical reader must question both the validity of the findings from the Michigan study and the reasons for discrepancies in results between the Jacobson and Rogan studies. Are the differences in neurobehavioral effects reported by the Jacobsons and colleagues, and Rogan and colleagues, due to the presence of confounders, exposure to different neurotoxicants, or subtle differences in methodologies? At present it is not possible to answer these questions. Nevertheless, certain commonalities exist between the Rogan and Jacobson studies, and most recently, the study conducted by Daly and colleagues in New York. All of these studies report alterations in the Brazelton Neonatal Behavioral Assessment Scale, suggesting that exposure to environmental contaminants (including PCBs) may induce subtle, transient alterations in maturation of the human CNS. Secondly, because contaminated fish contain a large number of putative developmental neurotoxicants (e.g., methyl-mercury, p,p'-DDE, PCBs, and pesticides), I am pessimistic that additional studies of human populations exposed to contaminated aquatic and marine fish and mammals will allow investigators to determine which contaminant(s) may be responsible for the observed association between fish consumption and neurobehavioral deficits. For example, although PCB body burdens have been measured in the majority of the epidemiological studies, PCBs may simply reflect exposure to other fish-borne contaminants. In light of the above statements, future epidemiological studies should focus on highly exposed susceptible populations such as occupationally exposed workers or the aged. Results from these studies would provide important information on the risk of perinatal or adult exposure to PCBs in susceptible populations, although generalization of results obtained in these populations to the general population may be fraught with difficulties. Finally, because of the limitations of epidemiological studies, particularly those studying fish-eating populations, future risk assessments should depend more heavily on laboratory derived data, including studies in nonhuman primates exposed to environmentally relevant mixtures and relevant doses of PCB congeners and other known or suspected neurotoxicants. PMID- 8725636 TI - Different congeners of PCBs/PCDFs may have contributed to different health outcomes in the Yucheng cohort. PMID- 8725637 TI - Sources and implications of interstudy and interindividual variability in the developmental neurotoxicity of PCBs. PMID- 8725638 TI - Critical issues for research on the neurobehavioral effects of PCBs in humans. PMID- 8725639 TI - Characterization of pregnancy outcome following thiram-induced ovulatory delay in the female rat. AB - A single injection of the dithiocarbamate fungicide, thiram, suppresses the proestrous surge of LH and delays ovulation for 24 h. In this study, we examined fertility after a thiram-induced delayed ovulation. Females were injected with thiram (50 mg/kg, IP) on proestrus (1300 h) and mated on the following evening. Control and thiram-treated, but nondelayed, females were injected and mated on the same day. The number of females in the thiram-delayed group that became pregnant was reduced and litter size on GD 20 was reduced: however, no obvious morphological anomalies were seen. The number of pregnant females and litter size was not altered in the thiram-nondelayed rats, indicating that it is the thiram induced delay in ovulation and not the exposure to thiram per se that was responsible for altered pregnancy outcome. On GD 7 and 11, the number of live fetuses per litter was reduced in the delayed females, but the number of implantation sites was not different from controls. On GD 11 the mean developmental score, head length, crown-rump length, and somite number in the delayed group were also reduced, indicating retarded development of live embryos. These results demonstrate that delayed ovulation induced by a single thiram exposure does not alter the number of oocytes released or the number that implant. However, the concept from these females are compromised during midgestation. PMID- 8725640 TI - Prenatal cocaine exposure and postnatal hypoxia independently decrease carotid body dopamine in neonatal rats. AB - The effects of prenatal cocaine exposure on the levels of carotid body dopamine (DA) and its metabolites 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) were investigated in 5-day-old rat pups exposed to normoxic and hypoxic conditions. Timed-pregnant Sprague-Dawley rats were injected b.i.d. with either cocaine HCl (30 mg/kg) or isotonic saline (1 ml/kg) from gestational days 7-21. On the fifth postnatal day, pups were subjected to either 20 min of 0.21 or 0.08 fractional inspired oxygen (FlO2). Under a strictly timed protocol, both carotid bodies were removed from each pup, placed in an antioxidant solution to prevent DA breakdown, and subsequently analyzed via HPLC with electrochemical detection to determine carotid body DA and DOPAC content. Two-way ANOVA revealed decreases in DA in cocaine-exposed pups. No HVA was detectable in any of the samples. The 0.08 FlO2 condition decreased DA compared to 0.21 FlO2. The additive consequences of DA depletion resulting from the combination of prenatal cocaine and postnatal hypoxia decreased carotid body DA to 14% of control levels, with several animals exhibiting DA content below detection limits. Considering the role of the carotid body in the ventilatory response to hypoxia, these data suggest that prenatal cocaine exposure may adversely affect the normal chemoreceptive function of the carotid body. PMID- 8725641 TI - Modification of acoustic startle reactivity by cocaine administration during the postnatal period: comparison with a specific serotonin reuptake inhibitor. AB - This study investigated whether exposure to cocaine during postnatal period affects the acoustic startle response (ASR) following administration of the serotonin (5-HT) agonists, 8-OH-DPAT and mCPP, in adulthood. To test the hypothesis that alterations in reactivity may be due to cocaine's effects at the 5-HT carrier, another group of rats was given fluoxetine, a specific 5-HT uptake inhibitor, during the same postnatal period and tested along with the cocaine treated rats. Male and female rats received 25 mg/kg/day cocaine HCl, fluoxetine HCl, or vehicle SC during postnatal days 11-20. At 75 days of age, subjects were ASR tested for 30 min on 2 consecutive days. On the first test day, there was a significant effect of treatment and gender with post hoc analysis indicating that, overall, the males were more reactive than the females and that the fluoxetine-treated males showed a pattern of reactivity resembling sensitization. On the second test day, subjects received a dose of the 5-HT1A agonist 8-OH-DPAT, the 5-HT1B/2C agonist, mCPP, or saline prior to being placed in the startle chamber. Cocaine-exposed males showed an enhanced response to 8-OH-DPAT and a reduction in the depression produced by mCPP administration compared to their response to saline. Fluoxetine exposed males showed a significant increase in startle response following saline administration compared to the rats receiving vehicle during the postnatal period and 8-OH-DPAT produced an insignificant enhancement of that startle response. mCPP reduced startle in fluoxetine-treated males as it did in the postnatal vehicle-treated controls. In females, the postnatal cocaine and fluoxetine treatments did not alter the response to 8-OH DPAT or mCPP compared to females receiving vehicle during the postnatal period. Together these data indicate that, in males, whereas postnatal cocaine alters the development of the 5-HT system as evidenced by an altered startle response to 5 HT agonists, cocaine does not produce the same alteration as that produced by the administration of a specific 5-HT uptake inhibitor during the same period of development. PMID- 8725642 TI - Characterization of the effects of N-hydroxy-IDPN on the auditory, vestibular, and olfactory systems in rats. AB - The mechanism of neurotoxicity of 3,3'-iminodipropionitrile (IDPN) has been widely debated, with either the parent compound or putative metabolites implicated in various studies. The N-hydroxylated form of IDPN (HO-IDPN) has been reported to cause the excitation with choreiform and circling (ECC) syndrome in rats at doses approximately one-eighth of that required to cause comparable signs in rats treated with IDPN. Because of the similarity of symptoms induced by HO IDPN and IDPN, we investigated the effect of HO-IDPN on other aspects of the nervous system affected by IDPN, specifically the auditory, vestibular, and olfactory systems. In addition, ECC symptoms were quantified to replicate the previous findings. HO-IDPN was administered ip in saline for 3 consecutive days to two different cohorts of young adult male Sprague-Dawley rats. The first cohort (60, 80, 100, and 120 mg/kg; n = 2/group, except for the 120 mg/kg group, where n = 1) was used in a dose range-finding study. After making the neurobehavioral assessments, animals were sacrificed for olfactory mucosal histopathology. Based on the outcome of the first study, the second cohort (n = 10/group) received saline or HO-IDPN at 100 mg/kg/day for 3 consecutive days. Two animals from each of these groups were sacrificed for olfactory mucosal histopathology; the remaining animals were tested for neurobehavioral effects 3 weeks after the last dose. Animals in the second cohort lost approximately 8% of their pretreatment body weight. All rats receiving the 100 mg/kg/day dose of HO IDPN (and the rat receiving 120 mg/kg/day) developed the ECC syndrome and signs of vestibular dysfunction within 4 days after the last dose. HO-IDPN caused a large decrease in the acoustic startle response and markedly elevated auditory thresholds at all frequencies tested. The threshold for the ECC syndrome and olfactory mucosal damage was 100 mg/kg. These studies extend previous findings on the neurotoxicity of HO-IDPN and point to the need for determining whether HO IDPN is an in vivo metabolite of IDPN. PMID- 8725643 TI - Effects of gestational and lactational exposure to TCDD or coplanar PCBs on spatial learning. AB - Recently we reported that in utero and lactational exposure to specific ortho substituted polychlorinated biphenyl (PCB) congeners resulted in a learning deficit on a delayed spatial alternation (DSA) task in female rats. In this study, spatial learning and memory was assessed following in utero and lactational exposure to coplanar PCBs or 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Time-mated Sprague-Dawley rats were dosed with PCB 77 (3,3',4,4' tetrachlorobiphenyl), 2 or 8 mg/kg/day; PCB 126 (3,3',4,4',5 pentachlorobiphenyl), 0.25 or 1.0 micrograms/kg/day; TCDD, 0.025 or 0.1 micrograms/kg/day; or corn oil vehicle via gavage on gestation days 10-16. Litters were culled to eight on day 2 and weaned on day 21. Beginning on day 80, one male and one female from each litter were tested on an eight-arm radial maze working memory task. The TCDD-exposed rats displayed pronounced decreases in errors relative to controls. PCB 77- and PCB 126-exposed rats showed similar, but less pronounced, decreases in errors. The same animals were later tested on a T maze DSA task, but no differences among groups were observed. In conclusion, perinatal exposure to low doses of TCDD or structurally related coplanar PCBs appeared to facilitate acquisition of a working memory task on the radial arm maze. This effect was very different from that previously observed in rats exposed to ortho-substituted PCB congeners. The rats exposed to ortho-substituted PCBs did not differ from controls on the radial arm maze and were impaired on the T-maze DSA task. Together these findings suggest that coplanar and ortho substituted PCBs may have different mechanisms of action on the CNS. PMID- 8725644 TI - Spatial acuity of touch: ubiquitous decline with aging revealed by repeated threshold testing. AB - Spatial acuity of touch, like that of vision, tends to decline eventually in nearly everybody's lifetime. This has been revealed by more thorough than customary testing of individual young and elderly subjects. Three kinds of acuity threshold were assessed repeatedly in the index finger. These measured ability to discriminate tactile (1) gaps (by a refined version of two-point threshold), (2) orientation of lines (across vs. along the finger), and (3) length of lines. These acuities relate to prominent discriminatory features of braille, and have been shown earlier to average about 1% larger per annum over the adult life span from about 20 to 80 years. Although there were reliable differences among the elderly subjects in the present experiment, all of them tested consistently worse than the least acute young adult controls. The customary single brief threshold tests heretofore applied are inadequate to capture this ubiquitous but differential individual deficit in advanced age; however, the average of six 15- to 20-min tests spread over 3 days proved more than adequate. The method of repeated threshold testing--applied earlier to olfactory and gustatory sensitivity, and now to tactile acuity--serves to dispel the notion that incidence of sensory loss with aging is highly idiosyncratic. PMID- 8725645 TI - Response of monkey glabrous skin mechanoreceptors to random noise sequences: II. Dynamic stimulus state analysis. AB - The responses of monkey glabrous skin slowly adapting (SAI, Merkel cell), rapidly adapting (RA, Meissner) and Pacinian corpuscle (PC) mechanoreceptors were analyzed as a function of the instantaneous position, velocity, and acceleration of a dynamic stimulus. For these experiments, a vibrotactile punctate stimulator was driven by a non-repeating-noise sequence. The resulting data (sampled stimulus waveform, windowed impulse) were processed in several ways. Initially, input-output correlation analysis was implemented to generate spike-aligned averages of the stimulus waveform preceding and following impulse initiation. From this analysis, it was determined that dual-responding RA and PC afferents- that is, those afferents that responded to both indenting and extracting stimulus movements--universally responded in a nearly perfectly symmetrical manner to the stimulus. Subsequently, two-dimensional (position, velocity) state histograms were generated and used to assess mechanoreceptor dynamic stimulus sensitivities. From these state histograms, it was determined that the threshold for impulse initiation by SAI afferents was preferentially sensitive to the indentation position of the stimulus, with only a minor sensitivity to stimulus velocity. RA afferent thresholds were sensitive to a continuum of dynamic stimulus velocities and positions. At the extremes, RA afferent impulses could be initiated by either a highly indented, low-velocity stimulus or high-velocity stimulus with a limited indentation position. PC afferents appeared to be preferentially sensitive to a combination of stimulus velocity and acceleration, but the data-sampling interval was too coarse to adequately resolve the full range of dynamic stimulus sensitivities. PMID- 8725646 TI - Sensory detection and pain thresholds in spinal cord injury patients with and without dysesthetic pain, and in chronic low back pain patients. AB - In an effort to understand the mechanisms involved in dysesthetic pain syndrome (DPS) in spinal cord injury (SCI) patients, four groups of 13 subjects each--SCI subjects with DPS, SCI subjects without pain, chronic low back pain subjects, and control subjects--were examined for sensory detection and pain thresholds at forearm, neck, and rostral trunk areas. Results indicated that the SCI pain group had significantly lower pain thresholds at all skin sites, compared to the SCI no pain and chronic low back pain groups, and at the rostral trunk skin site, compared to the control group. The SCI pain group also showed a lower sensory detection threshold at the rostral trunk skin site than did the SCI no-pain group. Equally important, the SCI no-pain group had detection and pain thresholds significantly higher than those of the control group. The results suggest fundamental differences in somatosensory processing when DPS is or is not a consequence of SCI. PMID- 8725647 TI - Rhythmical oral-motor activity recorded in an in vitro brainstem preparation. AB - The present study employed the neonatal rat isolated brainstem preparation to determine whether oral-motor rhythmical activity, a substrate for the complex behaviors of suckling and chewing, could be elicited in vitro by path application of excitatory amino acids (EAAs). Bath application of EAA agonists (kainate [KA], [+/-]-a-amino-3-hydroxy-5-methylisoxazole-4-propionic acid [AMPA], N-methyl-D, L aspartate [NMA]), in conjunction with the gamma-aminobutyric acid antagonist bicuculline, either failed to induce rhythmic activity (n = 17 preparations) or induced a low-amplitude, low-frequency burst discharge (< 1 Hz, n = 10 preparations) from the motor branches of the trigeminal nerves when the brainstem was contiguous from the spinomedullary junction to the superior colliculus. Burst activity was in most cases bilaterally synchronous. However, when a discrete coronal transection was made at the level of the facial colliculus, between the trigeminal and facial motor nuclei, the rhythmic bursts produced by the resultant 3- to 5-mm block of tissue following bath application of EAA agonists increased in amplitude and frequency (4-8 Hz, n = 35). Application of 6-cyano-7 nitroquinoxaline-2, 3-dione (CNQX), a non-N-methyl-D-aspartate (non-NMDA) receptor antagonist, blocked the rhythm induced by non-NMDA receptor agonist (n = 4) but was less effective in suppressing NMA-induced rhythmicity. In contrast, D, L-2-amino-5-phosphonovaleric acid (APV) blocked by both NMA-induced (n = 5) and, in most cases, KA-induced (n = 5) rhythmicity, suggesting an essential role for NMA receptors in production of EAA-induced rhythmical oral-motor activity in the neonatal rat. The present data demonstrate that a narrow, bilaterally distributed region of brainstem surrounding the trigeminal motor nucleus contains sufficient neuronal circuitry for the production of oral-motor rhythmogenesis. PMID- 8725648 TI - Mitochondrial distribution within the terminal neurite of the pacinian corpuscle. AB - Electron-microscopic analyses of the mitochondrial organization within the neurite innervating the Pacinian corpuscle (PC) were performed to test the hypothesis that the sites of mechanotransduction are the filopodia projecting from the neurite's surface. Since high concentrations of mitochondria imply the need for metabolic energy, and since transduction mechanisms are heavily dependent on such energy, it was reasoned that the greatest concentration of mitochondria should occur near the filopodia if they are involved in mechanotransduction. The analysis that the mitochondria lie close to the terminal neurite's membrane, on average within 0.4 microns, and thus are ideally located for supplying energy for membrane mechanisms. Although they can be found in a ring-like array, as seen in cross-sections of the terminal neurite, their greatest concentration occurs where the filopodia project from the terminal neurite. A linear algebraic analysis of the data set confirmed a high probability of the joint occurrence of a filopodial base and an increased frequency (number) of mitochondria. Thus the results provide further circumstantial support for the hypothesis that transduction within the PC mechanoreceptors takes place at or near the filopodia. PMID- 8725649 TI - A stereological analysis of the numerical distribution of neurons in dorsal root ganglia C4-T2 in adult macaque monkeys. AB - The numbers of neurons in dorsal root ganglia C4-T2 of adult monkeys (M. nemestrina) were estimated in celloidin-embedded material by means of the optical fractionator, a stereological procedures that combines the optical disector with a fractionator sampling scheme. On each side, counts of A-type, B-type, and total neurons were performed for the whole set of ganglia, as well as separately for ganglia C7, C8, and T1. Sampling and counting in this study were carried out with the help of an interactive computer system, the test grids being provided by the GRID general stereological software package (Olympus Denmark). The precision of the estimates for each animal was evaluated by computing the coefficient of error, which was kept at or below 0.10. The mean number of neurons on each side of the C4 - T2 set was 236,500 with a coefficient of variation among animals (CV) of 0.13. Of these neurons, 42% were A-type and 56% were B-type. Mean left-right differences among animals were below 1%, with low variability (CV = 0.07). The mean numbers of neurons in ganglia C7, C8, and T1 were, respectively, 46,000 (CV = 0.20), 51,000 (CV = 0.18), and 41,000 (CV = 0.22). Mean side differences for individual ganglia were 17%, 16%, and 12%, respectively, with high variability among animals. Intraanimal side differences were low for the whole set of ganglia (4%), as well as for the C7-T1 group (5%), but increased substantially when ganglia were considered separately (up to 17% on average in C7) or even in pairs of adjacent ganglia. These findings provide a quantitative frame for developmental or lesion studies in the peripheral somatosensory system of macaques, and warn against using single ganglia in studies requiring quantitative side comparisons. PMID- 8725650 TI - Somatotopic localization of thermal stimuli: I. A comparison of within- versus across-dermatomal separation of innocuous thermal stimuli. AB - Fourteen healthy subjects (4 males, 10 females) were asked to localize a thermal stimulus applied to the left distal forearm. Two temperature-controlled probes (1,2-cm2 area each) were strapped to the forearm, separated by 8 cm. The probes were positioned in one of three ways: (1) longitudinally within the C6 dermatome, (2) longitudinally within the C8 dermatomes, and (3) transversely with one probe within the C6 and one probe within the C8 dermatome. For any given stimulus configuration, cooling stimuli were localized significantly better than warming stimuli. For both warming and cooling, the transdermatomal configuration (transversely separated probes) provided significantly better localization than the intradermatomal configuration (longitudinally separated probes). Thus, the anisotropy that has been described for tactile spatial acuity is also present for thermal localization. These results suggest that cutaneous somatotopic information is integrated similarly for both and thermal stimuli, but differently within versus across dermatomes. PMID- 8725651 TI - The effects of aging on information-processing channels in the sense of touch: III. Differential sensitivity to changes in stimulus intensity. AB - Detection thresholds and difference limens were measured for 16 subjects ranging from 19 to 91 years of age. The stimuli were 250-Hz bursts of vibration applied through a 3.0-cm2 contactor to the thenar eminence of the right hand. Detection thresholds were higher in older than in younger subjects, as were the absolute values of difference limens. When the difference limen was expressed in relative terms as the proportion by which two stimuli had to differ in amplitude to be discriminated (delta alpha/alpha), discriminative capacities were unaffected by aging except for stimuli slightly above the detection threshold, in which case the limens of older subjects were significantly higher than those of younger subjects. The results are consistent with the hypothesis that elevations in the detection thresholds of older subjects are the results of reduced afferent input to central brain centers that, with regard to their capacity to detect the presence of threshold-level stimuli and to discriminate differences among suprathreshold stimuli, are relatively unaffected by aging. PMID- 8725652 TI - Advance directives for research: ethical problems and responses. PMID- 8725653 TI - The code of professional ethics of the Russian Society of Psychiatrists. Text and commentary. PMID- 8725654 TI - Parricide and mental illness. A study of 12 cases. PMID- 8725655 TI - Sexuality in the mental health system. Patients and practitioners. PMID- 8725656 TI - The Swedish forensic concept of severe mental disorder as related to personality disorders. An analysis of forensic psychiatric investigations of 1498 male offenders. PMID- 8725657 TI - Coercion and the outcome of psychiatric hospitalization. PMID- 8725658 TI - Simultaneous measurement of Ca2+ and pH by laser cytometry using fluo-3 and SNARF 1. AB - We present a new convenient method for simultaneous measurement of intracellular calcium concentration ([Ca2+]i) and intracellular pH (pHi) using laser cytometry with a mixture of fluo-3 (for [Ca2+]i) and SNARF-1 (for pHi), with iso excitation (488 nm)-dual emission (530 nm for fluo-3 and > 630 nm for SNARF-1). By using this technique, we measured the changes in [Ca2+]i and pHi in A-431 human epidermoid carcinoma cells and HSG human salivary gland cells stimulated by ATP. We found that alkalization in A-431 cell occurred with the elevation of [Ca2+]i; in contrast, alkalization in HSG cells did not occur at all, even though the elevation of [Ca2+]i was observed. Using BAPTA (a chelating agent of Ca2+) and amiloride (an inhibitor of the Na+/H+ exchanger), we found that the elevation of pHi requires the elevation of [Ca2+]i but that the elevation of [Ca2+]i does not always require a rise in pHi. From our results we conclude that elevation of [Ca2+]i takes precedence over the elevation of pHi in ATP-stimulated signal transduction. PMID- 8725659 TI - Flow cytometric analysis of the early phases of apoptosis by cellular and nuclear techniques. AB - The early events occurring during apoptosis at the plasma membrane, chromatin, and mitochondrial levels were investigated in freshly isolated irradiated human lymphocytes, growth factor-deprived cultured human lymphocytes, and dexamethasone (DEX)-treated murine thymocytes. In intact, unfixed cells, evaluation of the light scatter properties and of DNA stainability with ethidium bromide (EB) allowed a cell subset suggestive for initial apoptosis to be identified. The apoptotic nature of these cells was confirmed by cell sorting in irradiated human lymphocyte model. EB could not be substituted for by propidium iodide, indicating that the nature of DNA probe used is of major importance for detecting initial apoptotic changes. Because mitochondria are thought to represent a primary target during apoptosis, we measured the uptake of mitochondria transmembrane potential sensitive (Rhodamine 123) and nonsensitive (10-nonyl-acridine-orange) probes concomitantly with EB uptake. Cells starting apoptosis had an enhanced incorporation of both mitochondria dyes, which in combination with EB identified several cell subsets. This suggests that complex alterations in mitochondrial structure and functioning occur in the early stages of apoptosis. To investigate phenomena occurring at the chromatin level in similar phases of apoptosis, irradiated human lymphocytes and DEX-treated murine thymocytes were disrupted and DNA stainability assessed in nuclear suspensions. A transient increase in DNA stainability, i.e., the appearance of distinct hyperdiploid peaks in the human model and a generalised upward shift of the G0/1 peak in the murine model, was observed in the early phases of apoptosis concomitantly with specific alterations in light scattering properties. These findings suggest that chromatin texture is altered in early apoptosis and affects DNA stainability. PMID- 8725660 TI - Flow cytometric analysis of apoptosis and bcl-2 in primary breast carcinomas: clinical and biological implications. AB - Analysis of programmed cell death (apoptosis), of bcl-2, a critical regulator of this process, and of the proliferative fraction may provide detailed information on the biologic characteristics of tumor cell populations. To investigate the potential role of these parameters in assessing mammary carcinoma, we adapted flow cytometric procedures for concurrent measurement of apoptosis, bcl-2 expression, and cell proliferation in 54 primary breast carcinomas and correlated the findings with traditional clinicopathologic information. Overall, a significant inverse relationship between apoptosis levels and bcl-2 expression was observed (P = 0.005). Apoptosis levels correlated significantly with DNA aneuploidy (P = 0.03) and S + G2M fractions (P = 0.005) of these tumors. A significant correlation between bcl-2 expression and estrogen receptor positivity (P = 0.05) and DNA diploidy (P = 0.02) was noted. Bcl-2 expression, however, was inversely correlated with S + G2M fractions (P = 0.001). We conclude that analysis of apoptosis and bcl-2 by flow cytometry allows further characterization of tumor cell populations that may be useful for prognostic and therapeutic management of breast carcinoma. PMID- 8725661 TI - Quantification of apoptotic cells with fluorescein isothiocyanate-labeled annexin V in chinese hamster ovary cell cultures treated with cisplatin. AB - Plasma membrane binding of annexin V was used to detect and quantitate apoptotic cells induced by cytotoxic drug treatment in epithelial cell cultures. Chinese hamster ovary (CHO) cells were incubated for 2 h with the ID90 concentration of Cisplatin (20 microM), and 24, 48, 72, and 96 h later the unfixed cells were stained with fluorescein isothiocyanate (FITC)-conjugated annexin V. The fluorescence signal was quantitated by flow cytometry (FCM). During the early phase of the apoptotic response, the annexin V-binding frequency histograms showed two separate cell populations, a dimly and a brightly fluorescent one. At t = 96 h after drug incubation, when the process of apoptosis was completed, only the brightly fluorescent population was present. A dose-effect relationship could be established between the Cisplatin concentration used in the 2 h incubation and the binding of annexin V on the cell membrane, as estimated by FITC fluorescence. The dimly and brightly fluorescent populations were sorted on the basis of annexin V binding, and assayed for 1) DNA breaks by in situ nick translation assay and DNA content by DNA-propidium iodine fluorescence in a bivariate analysis, 2) membrane integrity by dye exclusion, and 3) morphological characteristics of apoptosis. The dimly fluorescent cell population appeared to represent apoptotic cells in the early phase of the death process, as demonstrated by intact cell membranes, normal DNA content, few DNA breaks, and chromatin condensation. The brightly fluorescent cells predominantly had sub-G1 DNA content, nuclear fragmentation, leaky cell membranes, and probably represent late apoptotic cells. These results demonstrate that cytotoxic drug-induced apoptosis can be quantitated by annexin V binding and that by using this assay early and late apoptotic cells can be identified. PMID- 8725662 TI - A novel assay to measure loss of plasma membrane asymmetry during apoptosis of adherent cells in culture. AB - Early during the process of apoptosis, cells lose their phospholipid membrane asymmetry and expose phosphatidylserine (PS) at the cell surface while maintaining their plasma membrane integrity intact. This process can be monitored for suspended cell types by using annexin V-FITC, which is a Ca(2+)-dependent, phospholipid-binding protein with high affinity for PS, and flow cytometry. If adherent cell types are to be studied for this apoptosis-associated phenomenon, then a problem is encountered, in that specific membrane damage occurs during harvesting. In this paper, a flow cytometric-based method is described that allows the measurement of loss of phospholipid asymmetry during apoptosis of adherent cells in culture. The method relies on the phospholipid binding property of biotinylated annexin V. Furthermore, the use of this conjugate allows tricolor flow cytometric analysis of apoptosis. Employing the method to MR65 cells, which were initiated by olomoucine to enter apoptosis, it is shown that PS exposure occurs early after the onset of apoptosis and, at the prevalent time-resolution, that PS exposure is accompanied by loss of both cytokeratin and DNA. The annexin V+ cells appear as a characteristic sub-G1 peak in the DNA histogram. PMID- 8725663 TI - New method of nuclear grading of tissue sections by means of digital image analysis with prognostic significance for node-negative breast cancer patients. AB - To optimize treatment of the individual patient with node-negative breast cancer, objective, reproducible, and standardized prognostic criteria are required. A number of factors have been studied in recent years, but until now it has been possible to obtain information about the risk of recurrence only for some patients belonging to subgroups with special characteristics. We report the establishment of an image analysis method for nuclear grading as an attempt to solve this problem. In a retrospective analysis, we used routine hematoxylin and eosinstained paraffin sections from 54 node-negative patients with surgery between 1980 and 1985. Cell scenes of primary tumors were scanned in a light microscope in successive focus positions to obtain three-dimensional information. After automatic image segmentation, nuclear features were calculated as input for a first binary classification tree to differentiate between tumor and nontumor cells. Tumor nuclei from patients with or without relapse were defined as high risk or low-risk nuclei, respectively, and were separated with a second tree. Feature values of the measured tumor nuclei from each patient were examined with this second tree to analyze whether the majority of nuclei for each patient were classified as high-risk or low-risk nuclei. Correct classification rates in the two binary cell classification trees were 88.0% and 83.8%, respectively. In the learning sample of our study, all patients with a relapse had the majority of nuclei in the high-risk group, most with more than 80%. Therefore, it seems to be possible to develop an image analytical risk profile system for nuclear grading to provide information on individual prognosis. PMID- 8725664 TI - Sample preparation and in situ hybridization techniques for automated molecular cytogenetic analysis of white blood cells. AB - With the advent of in situ hybridization techniques for the analysis of chromosome copy number or structure in interphase cells, the diagnostic and prognostic potential of cytogenetics has been augmented considerably. In theory, the strategies for detection of cytogenetically aberrant cells by in situ hybridization are simple and straightforward. In practice, however, they are fallible, because false classification of hybridization spot number or patterns occurs. When a decision has to be made on molecular cytogenetic normalcy or abnormalcy of a cell sample, the problem of false classification becomes particularly prominent if the fraction of aberrant cells is relatively small. In such mosaic situations, often > 200 cells have to be evaluated to reach a statistical sound figure. The manual enumeration of in situ hybridization spots in many cells in many patient samples is tedious. Assistance in the evaluation process by automation of microscope functions and image analysis techniques is, therefore, strongly indicated. Next to research and development of microscope hardware, camera technology, and image analysis, the optimization of the specimen for the (semi)automated microscopic analysis is essential, since factors such as cell density, thickness, and overlap have dramatic influences on the speed and complexity of the analysis process. Here we describe experiments that have led to a protocol for blood cell specimen that results in microscope preparations that are well suited for automated molecular cytogenetic analysis. PMID- 8725665 TI - Automation of spot counting in interphase cytogenetics using brightfield microscopy. AB - In situ hybridization techniques allow the enumeration of chromosomal abnormalities and form a great potential for many clinical applications. Although the use of fluorescent labels is preferable regarding sensitivity and colormultiplicity, chromogenic labels can provide an excellent alternative in relatively simple situations, e.g., where it is sufficient to use a centromere specific probe to detect abnormalities of one specific chromosome. When the frequency of chromosomal aberrations is low, several hundreds or even thousands of cells have to be evaluated to achieve sufficient statistical confidence. Since manual counting is tedious, fatiguing, and time consuming, automation can assist to process the slides more efficiently. Therefore, a system has been developed for automated spot counting using brightfield microscopy. This paper addresses both the hardware system aspects and the software image analysis algorithms for nuclei and spot detection. As a result of the automated slide analysis the system provides the frequency spot distribution of the selected cells. The automatic classification can, however, be overruled by human interaction, since each individual cell is stored in a gallery and can be relocated for visual inspection. With this system a thousand cells can be automatically analyzed in approximately 10 min, while an extra 5-10 min is necessary for visual evaluation. The performance of the system was analyzed using a model system for trisomy consisting of a mixture of male and female lymphocytes hybridized with probes for chromosomes 7 and Y. The sensitivity for trisomy detection in the seeding experiment was such that a frequency of 3% trisomic cells could be picked up automatically as being abnormal according to the multiple proportion test, while trisomy as low as 1.5% could be detected after interaction. PMID- 8725666 TI - Bull sperm head morphometry related to abnormal chromatin structure and fertility. AB - This study investigated the relationship between morphologically abnormal sperm, sperm chromatin structure, and fertility. Semen samples were obtained from 13 bulls. The sperm chromatin structure assay (SCSA), a sensitive measure of denaturability of sperm nuclear DNA in situ following acid treatment, was performed on each sample to quantitate abnormal chromatin structure. Feulgenstained sperm head morphology was measured by computerized image analysis (ONCOR-Image): Sixteen parameters were measured for each of 200 nuclei per sample. Fertility estimates were available for nine of the bulls. The SCSA variable SD alpha t was correlated with fertility ranking (r = 0.617, P < 0.01). No correlations were seen between means of the imaging variables and SCSA variables % COMP alpha t or SD alpha t. Significant correlations (P < 0.05) were seen between SD alpha t and the variation of imaging variables eccentricity, width, and light blobs. Significant correlations (P < 0.05) were seen between % COMP alpha t and the variation of imaging variables area, perimeter, p2a, bending energy, nmac, sphericity, eccentricity, length, and width. A regression model for fertility rankings incorporating the standard deviation of the imaging variables area, bending energy, nmac, eccentricity, condensity, light blobs, and dark blobs was highly significant (r2 = 0.999, P < 0.05). These results indicate that variation of morphometry measurements is likely a sensitive biomarker related to fertility potential and abnormal chromatin structure. PMID- 8725667 TI - Flow cytometric assessment of trophosphamide toxicity on mouse spermatogenesis. AB - The effects of trophosphamide on mouse reproductive cells have been investigated by flow cytometric analysis of testicular cell populations and alterations of sperm chromatin structure. Mice were treated with single intraperitoneal injections of TP, the doses ranging between 50 and 150 mg/kg, and were killed after 7, 14, 21, 28, 35, or 49 days. Dose-dependent reductions of tetraploid cells, round spermatids, and elongated spermatids were detected at 7, 21, and 28 days, respectively, reflecting cytotoxic damage to the differentiating spermatogonia compartment. The dose necessary to reduce the number of differentiating spermatogonia to half the control value was approximately 70 mg/kg. Stem cells were not affected by this treatment, and the normal spermatogenic process was restored after 7 weeks. In addition, cauda epididymal sperm were analyzed by the sperm chromatin structure assay, a flow cytometric measurement of the susceptibility of the sperm nuclear DNA to in situ acid denaturation; a statistically significant increase of sperm with altered chromatin structure was detected after a TP treatment of 150 mg/kg. Together with previous findings published in the literature, where the same doses induced heritable genetic damage, this study demonstrates a marked adverse cytotoxic effect of TP on the male reproductive integrity. All this information should be taken into consideration when TP is used in chemotherapeutic regimens. PMID- 8725668 TI - Fluorescence analysis of carrier rat and human erythrocytes loaded with FITC dextran. AB - Rat and human erythrocytes are inherently different with respect to slow dialysis encapsulation used in preparing carrier erythrocytes. The incorporation process, commonly measured with radioactive tracers, is always larger in human erythrocytes, mainly because the rat carrier cells are more fragile. When FITC Dextran (Dx) is used in the encapsulation process, and loaded rat and human RBCs are studied by fluorescence intensity, some additional events are evident. Not all cells of each population appear with a fluorescence signal, and not all show similar fluorescence intensity. Human RBCs show a higher percentage of marked cells and a higher fluorescence intensity than rat RBCs. Two populations, of high and low fluorescence, appear in FITC-Dx loaded rat erythrocytes. The human loaded RBCs show a similar peak distribution together with another peak in the middle scale of fluorescence. Therefore, a heterogeneity in the cell population as a result of the encapsulation process is manifested for both species. The fractionation of RBCs, loaded with either FITC-Dx or 125I-CA, by centrifugation on Ficoll-Paque reveals that the low density cells have much more substance incorporation than the counterpart cell subpopulation in the pellet. Therefore, the cell modifications produced by the encapsulation process are independent of the substance being incorporated. On the other hand, FITC-Dx, but not 125I-CA, shows a certain degree of association to RBCs membranes, especially in humans. PMID- 8725669 TI - Analysis of surface density of expression of molecules by flow cytometry. PMID- 8725670 TI - Physical activity assessment: a review of methods. AB - In order to determine the actual threshold level of habitual physical activity necessary to obtain protective effects against degenerative diseases, valid, reliable, nonreactive, and precise instruments are needed. Because regular, low intensity exercise provides prophylactic effects, the ability to distinguish activity patterns should be a primary consideration when choosing an instrument to assess activity in the field. Although self-report methods in the form of questionnaires are the instruments of choice when large populations are to be assessed, they are somewhat limited in their objectivity. The doubly labeled water (DLW) technique is a highly accurate and objective technique for assessing total daily energy expenditure that should be utilized more as a criterion measure for validating questionnaires that assess energy expenditure. However, the DLW technique remains too costly to be considered practical for large-scale studies. Supplementing questionnaires with a personal interview does elicit more detailed data, but activity monitors provide a more objective measure of activity that can be used as an adjunct to questionnaires. Several models of activity monitors are capable of collecting and storing data for many days, weeks, or even months. More importantly, the internal real-time clocks of these monitors allow the discrimination of activity patterns. Future studies should combine the use of these monitors with a suitable questionnaire. PMID- 8725671 TI - Bitterness in cheese: a review. AB - Bitterness, the necessary consequence of proteolysis, has been under investigation from different perspectives. This review attempts to give more up to-date information on the definition of some principal aspects, the relationship between the proteolytic activity and bitter peptide accumulation in cheese, and methods of isolation and detection of bitter peptides. Further knowledge on the physicochemical properties of bitter peptides in cheese as well as in synthetic peptides and the possible control methods for bitterness are discussed. Particular interest in using some strains of lactobacilli or their enzymes as an adjunct in accelerated ripened cheese (ARC) and enzyme-modified cheese (EMC) without bitterness is also described in detail. PMID- 8725672 TI - Cowpea flour: a potential ingredient in food products. AB - Cowpeas (Vigna unguiculata L. Walp) are an important grain legume in East and West African countries as well as in other developing countries. The U.S. is the only developed country producing large amounts of cowpeas. High protein (18 to 35%) and carbohydrate (50 to 65%) contents, together with an amino acid pattern complementary to that of cereal grains, make cowpeas potentially important to the human diet from a nutritional standpoint. Despite their potential as an inexpensive source of protein and energy, cowpeas are underutilized in the U.S. and other industrialized countries. A simple technology for preparing cowpea flour would stimulate increased consumption of this legume. This paper reviews the status of cowpea flour production technologies and uses of cowpea flour in various food products. PMID- 8725673 TI - Application of enzymes in food processing. AB - Enzymes offer potential for many exciting applications for the improvement of foods. There is still, however, a long way to go in realizing this potential. Economic factors such as achievement of optimum yields and efficient recovery of desired protein are the main deterrents in the use of enzymes. Changing values in society with respect to recombinant DNA and protein engineering technologies and the growing need to explore all alternative food sources may in time make enzyme applications more attractive to the food industry. Research is continuing on the commercially viable enzymes in use today to improve various properties such as thermostabilities, specificities, and catalytic efficiencies. New and unique enzymes continue to be developed for use in enzymatic reactions to produce food ingredients by hydrolysis, synthesis, or biocatalysis. An aggressive approach is needed to open new opportunities for enzyme applications that can benefit the food industry. PMID- 8725674 TI - Water activity, water glass dynamics, and the control of microbiological growth in foods. AB - Water is probably the single most important factor governing microbial spoilage in foods, and the concept of water activity (a(w)) has been very valuable because measured values generally correlate well with the potential for growth and metabolic activity. Despite some drawbacks (e.g., solute effect), the concept of a(w) has assisted food scientists in their effort to predict the onset of food spoilage as well as to control food-borne disease hazards in food products. In the last decade the concept of a(w) has been challenged. It has been suggested that reduced-moisture food products (e.g., low and intermediate) may be nonequilibrium systems and that most of them are in the amorphous metastable state, which is very sensitive to changes in moisture content and temperature. It has been proposed that the glass transition temperature Tg (temperature at which the glass-rubber transition occurs), is a parameter that can determine many product properties, the safety of foods among them. The concept of water dynamics, originating in a food polymer science approach, has been suggested instead of a(w) to better predict the microbial stability of intermediate moisture foods. The usage of a(w) to predict microbial safety of foods has been discouraged on the basis that (1) in intermediate-moisture foods the measured water vapor pressure is not an equilibrium one, and because a(w) is a thermodynamic concept, it refers only to equilibrium; and (2) the microbial response may differ at a particular a(w) when the latter is obtained with different solutes. This review analyzes these suggestions on the basis of abundant experimental evidence found in the literature. It is concluded that nonequilibrium effects (e.g., inability of water to diffuse in a semimoist food) appear to be in many cases slow within the time frame (food's shelf life) of the experiments and/or so small that they do not affect seriously the application of the a(w) concept as a predictor of microbial stability in foods. The claims that a food science polymer approach to understanding the behavior of aqueous sugar glasses and concentrated solutions may be used to predict the microbial stability of food systems is not substantiated by experimental evidence. This approach does not offer, at the present time, a better alternative to the concept of a(w) as a predictor of microbial growth in foods. It is also recognized that a(w) has several limitations and should be always used carefully, and this must include precautions regarding the possible influences of nonequilibrium situations. This aspect may be summarized by simply saying that anyone who is going to employ the term water activity must be aware of the implications of its definition. PMID- 8725675 TI - Dialyzers and dialysis membranes. Research directions. PMID- 8725676 TI - Has peritoneal dialysis peaked? The impact of the CANUSA study. The Canadian-USA Multicenter study. AB - We previously summarized evidence that an increase of dialysis dose is associated with better protein intake and better nutritional status. The CANUSA study suggests that higher dialysis doses are associated with better patient survival rates. It is imperative that we design prescriptions and techniques to provide greater dialysis doses compatible with a reasonable quality of existence. The reasons why U.S. dialysis patients may have higher mortality rates than in other countries for both hemodialysis and CPD may relate to factors other than dialysis adequacy and commonly appreciated morbidity factors; this needs further attention. PMID- 8725677 TI - Coronary revascularization in patients on dialysis. What treatment option should we choose? PMID- 8725678 TI - Use of two-dimensional and contrast echocardiography for venous cannula placement in venovenous extracorporeal life support. AB - The objective of this study was to determine the efficacy of two-dimensional and contrast echocardiograms to identify venous cannula position. Sequential sampling of 20 infants was evaluated by contrast echocardiography after meeting institutional criteria for extracorporeal life support. Each infant was placed on venovenous extracorporeal life support using a double-lumen cannula. After surgical placement was thought to be satisfactory, optimal two-dimensional images of the cannula were obtained via a subxiphoid or apical view and 2 ml agitated normal saline were injected rapidly into the nearest infusion port. Patient demographics and mixed venous saturations were noted. Distance of the venovenous cannula to tricuspid valve and distance of the venovenous cannula from the intra atrial septum was recorded. Echocardiograms were available for review on 18 of the 20 patients. Position of the venovenous cannula in relationship to the tricuspid valve was as follows: < 5 mm (8); 5-10 mm (5); > 10 mm (5). Mixed venous saturations decreased, which indicated less recirculation when the orientation of the tip of the cannula was toward the lateral wall in those who required repositioning. The authors conclude that two-dimensional and contrast echocardiography aid in the positioning of the venovenous cannula. Satisfactory position is approximately 5 mm from the tricuspid valve, with orientation toward the lateral wall of the right atrium. PMID- 8725679 TI - Anemia severity and missed dialysis treatments in erythropoietin-treated hemodialysis patients. AB - Neither the sociodemographic correlates nor the biochemical/clinical consequences of missed dialysis treatments have been well defined. During a 10 week period, the authors enumerated missed dialysis treatments among 430 patients randomly selected from a pool of 1,395 hemodialysis patients. A forward logistic regression model was used to determine whether a relationship existed between missed dialysis treatments and the following independent variables: age, gender, race, renal diagnosis, length of time on maintenance hemodialysis, co-morbidity index, modified Karnofsky score, employment status, household residents, and laboratory indices. Forty-three (10%) of 430 patients missed a total of 96 treatments. Despite equivalent treatment with erythropoietin, patients who missed dialysis treatment(s) had a lower mean hematocrit (27 +/- 4.3%) at the end of the study than those patients who underwent all treatments (29 +/- 4.5%) (p = 0.0287). Mean serum albumin and creatinine levels were equivalent in compliant and noncompliant patients. Recent starts (p = 0.0048), and younger patients (p = 0.0424) were most likely to miss dialysis treatment(s). One of the major consequences of missed dialysis treatment(s) is exacerbation of anemia, and younger patients and freshly started patients are more likely to miss scheduled dialysis treatments than their respective counterparts. PMID- 8725680 TI - Gastrointestinal consequences of left ventricular assist device placement. AB - Left ventricular assist devices effectively improve hemodynamic function and reverse renal and hepatic dysfunction; however, their effects upon the gastrointestinal (Gl) system have not been addressed. We evaluated Gl function in 27 left ventricular assist device recipients using interviews, Gl contrast studies, endoscopy, and 99mTc sulfur colloid studies of esophageal transit and gastric emptying. While on left ventricular assist device support (mean duration of 84 days), 19 patients reported early satiety and/or nausea, and 1 was unable to tolerate oral intake. Esophageal transit time (normal, < 10 sec) was borderline slow at 14 +/- 4 (mean +/- standard error of the mean) and gastric emptying (normal < 90 min) was prolonged (range of 106-506 min, mean = 283 +/- 69 min). In a 1-38 month follow-up, gastric function subjectively improved in all. Six patients had intraperitoneal device placement. One died of aspiration pneumonia secondary to small bowel obstruction, and one had prolonged inability to tolerate oral intake, which required feeding jejunostomy tube placement. The 21 patients with pre peritoneal placement of the device did not require Gl operative interventions and had no catastrophic Gl events; they had mild to no Gl complaints. Pre peritoneal placement may mitigate early satiety and obviate serious Gl complications. PMID- 8725681 TI - Identification of peak stresses in cardiac prostheses. A comparison of two dimensional versus three-dimensional principal stress analyses. AB - This study assessed the accuracy of using a two-dimensional principal stress analysis compared to a three-dimensional analysis in estimating peak turbulent stresses in complex three-dimensional flows associated with cardiac prostheses. Three-component, coincident laser Doppler anemometer measurements were obtained in steady flow downstream of three prosthetic valves: a St. Jude bileaflet, Bjork Shiley monostrut tilting disc, and Starr-Edwards ball and cage. Two-dimensional and three-dimensional principal stress analyses were performed to identify local peak stresses. Valves with locally two-dimensional flows exhibited a 10-15% underestimation of the largest measured normal stresses compared to the three dimensional principal stresses. In nearly all flows, measured shear stresses underestimated peak principal shear stresses by 10-100%. Differences between the two-dimensional and three-dimensional principal stress analysis were less than 10% in locally two-dimensional flows. In three-dimensional flows, the two dimensional principal stresses typically underestimated three-dimensional values by nearly 20%. However, the agreement of the two-dimensional principal stress with the three-dimensional principal stresses was dependent upon the two velocity components used in the two-dimensional analysis, and was observed to vary across the valve flow field because of flow structure variation. The use of a two dimensional principal stress analysis with two-component velocity data obtained from measurements misaligned with the plane of maximum mean flow shear can underpredict maximum shear stresses by as much as 100%. PMID- 8725683 TI - Performance assessment of prosthetic heart valves using the energy index method. AB - Traditional methods of characterizing valvular performance use some estimation of the effective opening area and the percent regurgitant volume. These methods are cumbersome because two parameters are used and their importance relative to one another is not revealed. The authors propose the use of a single parameter that is physically meaningful and accounts for characteristics of the valve throughout the cardiac cycle. The energy index, derived with use of a phase-by-phase analysis of the cardiac cycle, describes the energetic efficiency of the valve. The method's final form is: [Formula: see text] where Eps is the hydraulic energy available after systole, E+ is the energy dissipated in the valve while flow is positive, [Symbol: see text]_ and [Symbol: see text] are the regurgitant and forward volumes, respectively. Use of the El requires on-line measurement of valvular flow rate and pressure drop. The El was applied to a Medtronic-Hall (Minneapolis, MN), 25 mm prosthetic valve mounted in the aortic position of a cardiovascular simulator. Mild and severe degrees of valvular stenosis and regurgitance were simulated. Results indicate that the El is sensitive to either valvular condition and remains nearly constant, at 87%, for the normal valve tested over cardiac rates ranging from 50 to 100 beats per minute. PMID- 8725682 TI - The impact of co-morbid risk factors at the start of dialysis upon the survival of ESRD patients. AB - By using a computerized database, we have catalogued the presence of 29 co-morbid risk factors in 683 patients with end-stage renal disease who started dialysis from 1970 through 1989, with follow-up through 1992. The authors hypothesized that current end-stage renal disease patients have more serious co-morbid risk factors impacting upon their mortality rate. Quantitation of dialysis patient co morbidity, as a measure of patient illness, is lacking in the general nephrology literature. Seven co-morbid risk factors have been reserved for new dialysis patients: hypertension, low albumin, cerebral vascular disease, peripheral vascular disease, pre-existing cardiac disease, abnormal EKG/old myocardial infarction, and congestive heart failure. Except for low serum albumin, the proportion of patients with the six other co-morbid risk factors has increased significantly over this 20-year period (p < 0.0001, chi-square test for hypertension, peripheral vascular disease, pre-existing cardiac disease, abnormal EKG/old myocardial infarction, and congestive heart failure, and p < 0.006 for cerebral vascular disease). In addition, the co-morbid risk factors of hypertension, low serum albumin, and pre-existing cardiac disease at the start of dialysis were strongly prognostic of survival. The Cox proportional hazards regression model identified these three risks, among other factors, that were significantly associated with a decreased survival, with risk ratios ranging from 1.40-1.66. These results support the authors' hypothesis that incoming end-stage renal disease patients, who recently start dialysis, are sicker than in the earlier years of the authors' program. If the authors' patients reflect the national end-stage renal disease population, the presence of co-morbid risk factors may, in part, explain the continuing high mortality of dialysis patients. PMID- 8725684 TI - Clinical experience with an anatomic snuff box arteriovenous fistula in hemodialysis patients. AB - The present study was performed to evaluate the clinical usefulness of an anatomical snuff box arteriovenous fistula with special reference to its short term and long-term patency in 139 hemodialysis patients. A snuff box fistula was established in 139 patients, including 39 with diabetic nephropathy (DN group) and 100 with non diabetic nephropathy (non DN Group). Fistula blood flow was measured by Doppler ultrasound in 18 patients. Early and late obstruction was observed in 24 (17%) of the 139 patients. The long-term patency rate of the snuff box fistula in the non DN group was 87% at 57 months, whereas the patency rate at 57 months in the DN group (72%) was significantly (p < 0.05) lower than that in the non DN group. The increase in arterialized vein blood flow in DN patients was less marked than that in the non DN patients. These results suggest that the high prevalence of late obstruction in DN patients may partly be reflected by an insufficient increase of arterialized vein blood flow at the snuff box site. PMID- 8725685 TI - Modeling of oxygen uptake in perfluorocarbon emulsions. Some comparisons with uptake by blood. AB - The use of perfluorocarbons emulsified in water as blood substitutes (artificial blood), is well known. Although considerable research has been devoted to the study of stability, toxicity, and gas solubility properties of these emulsions, there is no quantitative guide to the oxygen transport behavior in such emulsions, especially with reference to this transport process in actual blood. This paper describes a mathematical model from which the oxygen flux into a straight, cylindrical tube carrying a perfluorocarbon emulsion may be computed. The solutions to the proposed model can be adapted to that for a capillary or for a single tube in a blood oxygenator. The rates of oxygen transfer, so obtained, have been compared with analogous transfer rates that can be achieved in natural blood flowing under identical conditions. Therefore, the minimal solubilization capacity for oxygen required of a perfluorocarbon emulsion can be estimated on a quantitative basis. The modeling approach used in this study is based on the well tested theory of mass transfer in microheterogeneous media reported in the chemical engineering literature. PMID- 8725686 TI - Experimental study of extraaortic balloon counterpulsation as a bridge to other mechanical assists. AB - A special extraaortic balloon was developed that can be placed around the ascending aorta. This balloon can easily support the heart temporarily in a median sternotomy field, especially in cases in which it is difficult to use intraaortic balloon pumping because of peripheral arterial disease. The goal of this study was to judge the applicability of this extraaortic balloon counterpulsation. An extraaortic balloon was placed around the ascending aorta of eight adult canines. Two heart failure models were used in this study: group A moderate heart failure; group B-severe heart failure. In group A, the aortic systolic pressure was significantly reduced (9.3%, p < 0.01), but in group B, there was no significant change. In group A, there was a significant increase in cardiac output (12.0%, p < 0.01), but in group B, there was no significant change. The endocardial viability ratio in both groups significantly increased (group A: 11.3%, p < 0.01; group B: 11.9%, p < 0.05). An extraaortic balloon around the ascending aorta is easily applicable through a median sternotomy, and can be used as a bridge to more powerful mechanical assists when intraaortic balloon pumping cannot be used. PMID- 8725688 TI - Advances in the management of respiratory failure. Aqueous preparations of oxygen. PMID- 8725689 TI - Advances in the management of respiratory failure. Nitric oxide. PMID- 8725690 TI - Advances in the management of respiratory failure. Accelerating lung growth. PMID- 8725691 TI - Advances in the management of respiratory failure. Advanced strategies for mechanical ventilation in severe acute respiratory failure. AB - A number of strategies exist to improve gas exchange during mechanical ventilation. Any strategy used, however, should have as a basic tenet of its use the principles of lung rest, the reduction of the risk of ventilator associated lung injury, and avoidance of hemodynamic compromise. The techniques presented above are intended to meet these goals and represent measures that can be applied without significant additional resources. PMID- 8725692 TI - Advances in the management of respiratory failure. Intracorporeal membrane gas exchange: will it still prevail? PMID- 8725693 TI - Advances in the management of respiratory failure. Liquid ventilation in the setting of respiratory failure. PMID- 8725694 TI - Results of mechanical ventricular assistance for the treatment of post cardiotomy cardiogenic shock. AB - The voluntary submission of data on patients who receive mechanical circulatory assistance for post cardiotomy cardiogenic shock and acute myocardial infarction has resulted in an established data base for the ongoing evaluation of these devices. Since its inception in 1985, there have been 1,279 cases reported of implanted ventricular assist devices for post cardiotomy cardiogenic shock. The average duration of support for these devices is approximately 4 days. Those patients who underwent implantation of a centrifugal pump had significantly shorter periods of support when compared with the pneumatically supported group. Patients had similar periods of support when comparing clinical outcome. Approximately 70% of the reported cases underwent centrifugal assistance. The majority of reported cases were supported with isolated left ventricular assistance. There was no significant difference in overall outcome statistics when comparing type or mode of support. Approximately 45% of reported patients were weaned from circulatory assistance, and 25% of all patients survived to discharge. These numbers remain consistent with previously reported statistics. The results with acute myocardial infarction remain limited. Of the 96 reported patients, 26% were weaned from support and only 11.5% survived to discharge. This report continues to support the use and research for advancement of these devices toward the treatment of post cardiotomy cardiogenic shock. PMID- 8725696 TI - The use of a second course of extracorporeal membrane oxygenation in neonatal patients. AB - Extracorporeal membrane oxygenation (ECMO) has been successful in rescuing near term or term infants in cardio-respiratory failure that results from a reversible disease process. In most cases, only one course of ECMO is needed to save these infants. However, a second course of ECMO may be beneficial in a select group of infants when recurrent persistent pulmonary hypertension develops. Other than abstract form, this is the first report of the use of a second course of ECMO in the literature. The authors report on three infants, two with recurrent persistent pulmonary hypertension secondary to congenital diaphragmatic hernia and one with necrotizing tracheobronchitis after Group B streptococcal sepsis who were treated at their institution with a second course of ECMO. Technical considerations in using a second course of ECMO depend upon the initial vessel cannulation site, time elapsed between cannulations, and the condition of the original artery and vein. By adopting a stenting procedure in those infants whose initial trial off was equivocal, a second cannulation may be prevented in neonatal patients with recurrent persistent pulmonary hypertension. PMID- 8725695 TI - National surveillance of dialysis associated diseases in the United States, 1993. AB - To determine trends in a number of hemodialysis associated diseases and practices, the Centers for Disease Control and Prevention, in collaboration with the Health Care Financing Administration, performed a mail survey of 2,304 chronic hemodialysis centers in the United States in 1993. By the end of 1993, at least three doses of hepatitis B vaccine were administered to 29% of patients and 76% of staff at responding centers. Hepatitis B surface antigen was present at low frequency in patients (incidence = 0.1%, prevalence = 1.2%) and staff members (incidence = 0.2%, prevalence = 0.3%). The 1993 incidence of hepatitis B virus infection among patients was higher at centers that accepted hepatitis B surface antigen positive patients but did not use a separate room and dialysis machine for treatment of these patients, government and profit (versus nonprofit) centers, and centers in four End Stage Renal Disease Networks. The prevalence of antibody to hepatitis C virus was 9.7% among patients and 1.6% among staff members. Pyrogenic reactions in the absence of septicemia were reported by 21% of centers and associated with use of high flux dialysis. Human immunodeficiency virus infection was known to be present in 1.5% of patients; 34% of centers reported providing hemodialysis to one or more human immunodeficiency virus infected patients. PMID- 8725697 TI - Prolonged artificial liver support in a child with fulminant hepatic failure. AB - In Japan, liver transplantation from brain dead donors has not yet started. The authors present the first report of a clinical experience with a child with fulminant hepatic failure in whom the combined treatment of plasma exchange and continuous hemodiafiltration using a high-performance polymethylmethacrylate membrane was used successfully to sustain life for a period of as long as 54 days before liver transplantation from a living donor could be performed. The combination of plasma exchange and continuous hemodiafiltration appeared to maintain blood coagulation and level of consciousness effectively. Although the combined use of plasma exchange and continuous hemodiafiltration is still unsatisfactory as an artificial liver support, the authors suggest that this technique may be useful to support the life of a child who awaits liver transplantation. PMID- 8725698 TI - Successful use of extracorporeal membrane oxygenation in the treatment of acute chest syndrome in a child with severe sickle cell anemia. AB - Extracorporeal membrane oxygenation (ECMO) is widely used in the treatment of respiratory and cardiovascular failure in neonatal patients. The authors present a case of a child with hemoglobin SS disease who was treated with ECMO after acute chest syndrome and acute respiratory distress syndrome developed. They also present data from the Extracorporeal Life Support Organization on this use of ECMO from other centers. To date, there have been 15 pediatric patients with acute chest syndrome treated with ECMO. Survival rate has been 26%. In selected patients with severe disease, ECMO can provide support at a lower mean airway pressure, allow for aggressive pulmonary lavage, and maintain adequate tissue oxygen delivery until the patient is more stable. Patients who might benefit include those with poor ventilation secondary to mucous plugging and barotrauma. The best success with these patients might be anticipated from venoarterial ECMO. Patients with severe cardiac or neurologic deterioration may constitute a group less likely to survive. PMID- 8725699 TI - Thromboembolic complications in a peripartum cardiomyopathy patient supported with the Abiomed BVS-5000 ventricular assist device. AB - A case of systemic embolization arising from an akinetic left ventricle in a peripartum cardiomyopathy patient supported with the Abiomed BVS-5000 ventricular assist device is reported. In an akinetic ventricle, prevention of blood stasis by positioning the atrial cannula tip across the mitral valve to decompress the ventricle may prevent this complication. PMID- 8725700 TI - Pharmacokinetics of depot medroxyprogesterone acetate contraception. AB - Depot medroxyprogesterone acetate (DMPA) is an aqueous suspension of 17-acetoxy 6 methyl progestin administered by intramuscular injection for long-term contraception. This highly effective injectable formulation of medroxyprogesterone acetate (MPA) has a prolonged duration of action since the progestin is released slowly from the muscle. MPA is detected in the serum within 30 minutes after an injection of 150 mg. Serum concentrations vary between individual women but generally plateau at about 1.0 ng/mL for about three months, after which there is a gradual decline. In some women, MPA can be detected in the serum for as long as nine months after a single injection of 150 mg. The circulating MPA initially inhibits the midcycle leutinizing hormone (LH) peak, but LH and follicle stimulating hormone (FSH) levels remain in the range of those for the luteal phase of a pretreatment control cycle. Since ovulation is inhibited, serum progesterone levels remain low (< 0.4 ng/mL) for several months following an injection of DMPA. When MPA levels fall below 0.1 ng/mL, ovulation resumes. Thus, return to fertility is delayed for several months if a woman wishes to conceive after receiving one or more injections of DMPA. Following an injection of DMPA, serum estradiol levels initially are in the early to midfollicular phase range (mean approximately 50 pg/nL). Serum estradiol levels begin to rise about four months after a single injection when MPA levels fall below 0.5 ng/mL. For women who have used DMPA for several years, serum estradiol levels range between 10 and 92 pg/mL, with mean levels of about 40 pg/mL. Despite these low levels of estradiol, hot flushes are a rare event, and the vaginal epithelium remains moist and well rugated. Women using DMPA for several years do not observe a change in breast size. DMPA causes the endometrium to become atrophic, with small, straight endometrial glands and decidualized stroma. The cervical mucus remains thick and viscid. DMPA is a very effective form of contraception because of its multiple mechanisms of action and slow release into the circulation. PMID- 8725701 TI - Counseling issues and management of side effects for women using depot medroxyprogesterone acetate contraception. AB - Patients satisfaction is crucial to maximizing long-term utilization and efficacy of any contraceptive method. Satisfaction is enhanced when appropriate preutilization counseling is offered and when side effects are successfully managed. This article provides a conceptual model for patient counseling, highlights the significant points that should be included in counseling patients about depot medroxyprogesterone acetate (DMPA) and offers clinical suggestions to help evaluate and treat the more common side effects associated with DMPA use. PMID- 8725702 TI - Depot medroxyprogesterone acetate contraception. Metabolic parameters and mood changes. AB - This review summarizes the existing literature regarding metabolic parameters and mood changes in women using depot medroxyprogesterone acetate (DMPA). MEDLINE and additional bibliographic sources were searched to identify English-language articles describing clinical trials and cross-sectional studies of DMPA published during the last 20 years. DMPA has little or no effect on glucose tolerance, but insulin levels may increase. An increase in low-density lipoprotein cholesterol and a decrease in high-density lipoprotein cholesterol have been observed in cross-sectional and longitudinal studies; however, there are no data to indicate whether the observed lipid alterations lead to adverse clinical events. There is some evidence suggesting decreased bone density in long-term DMPA users. Longitudinal data regarding bone density changes are needed to assess changes both during and after use. Most cross-sectional and longitudinal studies show increased mean weight or weight gain in DMPA users. Controlled studies of weight change that evaluate subgroups and effects of confounding variables are also needed to provide constructive advice to patients. Data regarding depression or mood changes in DMPA users are scanty and do not support a causal relationship between use of this contraceptive and affective disorders. For all of these parameters, case reports indicating adverse clinical outcomes are nearly absent from the literature despite widespread long-term international use of DMPA. PMID- 8725703 TI - Use of depot medroxyprogesterone acetate contraception in adolescents. AB - More than 1 million teen pregnancies occur each year in the United States. DMPA offers several distinct advantages over other contraceptive methods in adolescents. The duration of effectiveness and convenience of DMPA are appealing to teens. Most teens who are current users view amenorrhea as a positive feature of DMPA. Although most young women are able to tolerate the irregular bleeding, some find the cycle disruption concerning. Satisfaction rates among adolescent DMPA users is high. PMID- 8725704 TI - Depot medroxyprogesterone acetate contraception in women with medical problems. AB - Since few clinical trials of contraceptive agents are conducted on women with medical problems, use of these agents in these populations is often based more upon theoretical considerations than clinical data. Clinicians must distinguish the risk of estrogen in the combined oral contraceptive from the risk of the progestin-only contraceptive methods. This review compares the risks of pregnancy with the risks of contraceptive methods for patients with chronic hypertension, cardiac disease, thrombotic disorders, diabetes, epilepsy, lupus erythematosus and other medical disorders. For women with certain medical problems, estrogen, but not progestogen, may be contraindicated. For these women, a long-acting progestogen, such as depot medroxyprogesterone acetate (DMPA), may offer distinct advantages. PMID- 8725705 TI - Depot medroxyprogesterone acetate contraception and the risk of breast and gynecologic cancer. AB - As the use of depot medroxyprogesterone acetate (DMPA) contraception increases in the United States and worldwide, assessing any impact on the risk of breast and gynecologic cancers becomes increasingly important. Overall, use of DMPA does not affect breast cancer risk. An elevated risk of this disease, however, has been noted in recent or current users. Hence, the association between DMPA use and breast cancer risk appears similar to that observed with oral contraceptives. Use of DMPA is associated with an 80% risk reduction of endometrial adenocarcinoma, a level of protection even greater than that noted with oral contraceptives. The risk of epithelial ovarian cancer and cervical neoplasia does not appear to be affected by DMPA use. Except for a substantial reduction in the risk of endometrial adenocarcinoma, use of DMPA does not appear to influence reproductive tract cancer risks. PMID- 8725706 TI - Noncontraceptive benefits and therapeutic uses of depot medroxyprogesterone acetate. AB - This paper reviews the noncontraceptive benefits and therapeutic uses of depot medroxyprogesterone acetate (DMPA). Relevant articles were reviewed using a computerized Medline search of the literature from 1966 to 1995. Good evidence shows that DMPA use is associated with reduced iron-deficiency anemia, protection against pelvic inflammatory disease, protection from endometrial cancer and improved hematologic parameters among users with sickle cell disease. More studies are needed to fully assess DMPA's impact on other disorders. PMID- 8725707 TI - Factor V (Arg 506-->Gln) mutation in young survivors of myocardial infarction. AB - Many young patients with venous thromboembolic disease are partially resistant to the anticoagulant action of activated protein C as a result of factor V (Arg 506 -> Gln) mutation. The frequency of this mutation in young patients with arterial thrombotic diseases, such as myocardial infarction, is less well established. We studied 100 young patients with myocardial infarction and 100 age- and sex matched controls. One patient (1%; 95% CL 0.05-6.2) and two controls (2%; 95% CL 0.3-7.7) were heterozygotes for the mutation; there was no homozygote in either group. Hence, premature myocardial infarction is not associated with heterozygosity for factor V (Arg 506 --> Gln) mutation. PMID- 8725708 TI - Heightened thrombin generation in individuals with resistance to activated protein C. AB - We chose to evaluate whether or not a state of biochemical hypercoagulability was present in 74 individuals (69 heterozygotes and 5 homozygotes) resistant to activated protein C (APC) due to the Arg506 --> Gln mutation in the factor V gene. To this end, plasma levels of two markers of thrombin formation, prothrombin fragment 1 + 2 (F1+2) and thrombin-antithrombin complexes (TAT) were measured. High levels of F1+2 and TAT were found in 32% and 23% of APC-resistant individuals vs 4% in controls. The levels of these markers tended to be particularly elevated in three homozygous subjects. A significant positive correlation between F1+2 and TAT was present in APC-resistant individuals. No relationship between marker values and the previous occurrence of thrombotic episodes was found. Therefore, by measuring F1+2 and TAT a state of biochemical hypercoagulability has been identified in about one-third of APC-resistant individuals. This frequency is similar to that previously observed in comparable individuals with inherited deficiencies of protein C and protein S, which are usually associated with a stronger thrombotic tendency than APC-resistant individuals. PMID- 8725709 TI - Comparison of two warfarin regimens in the prevention of venous thrombosis following total knee replacement. AB - A prospective, randomized trial was conducted to compare the effectiveness and safety of warfarin given in two regimens in prevention of venous thrombosis after total knee replacement. Adult patients scheduled for primary or revision total knee replacement were randomly assigned to receive either a "two-step" warfarin regimen beginning 10-14 days pre-operatively or, alternatively, to begin warfarin the night before surgery. Post-operatively, the dose was adjusted in both groups to achieve a target International Normalized Ratio (INR) of 2.2 and prophylaxis was continued until venography on post-operative days five through nine. Bleeding was assessed by surgical blood loss, transfusion requirements, changes in hematocrit, and clinically identified bleeding complications. The occurrence of deep vein thrombosis was nearly the same in the two treatment groups, 39% in patients randomized to the two-step regimen as compared to 38% in those beginning the night before surgery. The occurrence of proximal vein thrombosis was also similar, 5% versus 7% (p = NS). Patients in the two-step group received 1.33 +/- 1.26 transfusions compared to 0.95 +/- 1.22 in the night before group (p < 0.05) and also had a lower nadir post-operative hematocrit of 26.7 +/- 3.1 as compared to 28.5 +/- 3.2 (p < 0.0001). Major bleeding complications were associated with excessively prolonged INRs and occurred in five patients in the two-step group and two in the night before group. Patients in both groups who developed thrombosis had a significantly lower INR on post-operative days two and three compared to those without thrombosis. We conclude that a prophylactic warfarin regimen for prevention of deep vein thrombosis after total knee replacement beginning the night before surgery is more convenient and may be associated with less bleeding than a regimen beginning warfarin 10-14 days pre-operatively. Careful control of anticoagulant intensity is needed to achieve maximum effectiveness and avoidance of bleeding complications. PMID- 8725710 TI - Hypercoagulable state in patients with Takayasu's arteritis. AB - Thrombus formation is recognized pathologically in the affected arteries and is supposed to play a major role in the pathogenesis of Takayasu's arteritis; however, hemostatic conditions in this disorder have not been elucidated fully. We determined plasma levels of molecular markers for platelet activity (platelet factor 4; PF4, beta-thromboglobulin; beta TG), thrombotic status (thrombin antithrombin III complex; TAT, fibrinopeptide A; FPA), fibrinolytic status (plasmin-alpha 2-plasmin inhibitor complex; PIC, D-dimer), and endothelial injury (von Willebrand factor antigen; vWF:Ag, thrombomodulin; TM) in 30 patients with Takayasu's arteritis and 20 age-matched control subjects. Plasma levels of PF4, beta TG, TAT, FPA and D-dimer, but not PIC, in patients with Takayasu's arteritis were substantially higher than those in normal control subjects. The levels of these markers were not different between the active and inactive stages of the disease. Plasma levels of vWF:Ag in patients with Takayasu's arteritis did not differ significantly from those in normal subjects, and plasma levels of TM were significantly lower than those in normal subjects. In patients with Takayasu's arteritis, platelet and coagulation activities are significantly increased, leading to hypercoagulable state and thrombus formation, although there is little, if any, endothelial damage. PMID- 8725711 TI - Reactivity to beta 2 glycoprotein I clearly differentiates anticardiolipin antibodies from antiphospholipid syndrome and syphilis. AB - Anticardiolipin antibodies (aCL) detected by standard ELISA are found in association with autoimmune and infectious diseases. It is now recognized that beta 2 glycoprotein I (beta 2GPI) is a cofactor for aCL binding to cardiolipin (CL). To examine differences in cofactor requirements, aCL positive sera from patients with the antiphospholipid syndrome (APS) and syphilis were studied. Using an ELISA with human purified beta 2GPI adsorbed onto irradiated plates, we detected high binding activity in 29 out of 35 samples from APS patients and low in only 1 out of 37 aCL positive syphilis sera. Moreover, a good correlation (r = 0.79) was also observed in the former group between aCL and anti beta 2GPI. Whole IgG and affinity purified IgG aCL from APS patients did not bind to CL in the absence of beta 2GPI, but recognized beta 2GPI on irradiated plates in the absence of phospholipids. In contrast, IgG purified from syphilis patients only bound to CL alone. Taken together, these data indicate that performing both ELISA (aCL and anti beta 2GPI) it could be possible to distinguish aCL from autoimmune or infectious diseases. PMID- 8725712 TI - Autoantibodies to phospholipid-binding plasma proteins in patients with thrombosis and phospholipid-reactive antibodies. AB - Anti-phospholipid (aPL) antibodies are defined as antibodies detected in systems employing phospholipids (PL). This general definition is misleading as it comprises a large group of autoimmune phospholipid-reactive antibodies that are directed against specific phospholipid-binding plasma proteins, such as beta 2 glycoprotein I (beta 2GPI) and prothrombin. Definition of phospholipid-reacting antibodies according to the plasma protein against which they are directed appears more appropriate and could be useful in understanding clinical events and pathogenic mechanisms. Using ELISA systems we have studied the presence of antibodies directed against specific phospholipid-binding proteins in a series of 22 patients with thrombosis and phospholipid-reactive antibodies of the IgG isotype. High levels of anti-beta 2 GPI IgG were detected in all 22 patients. Normal values were calculated on the basis of OD values at 405 nm (OD405) obtained for 22 age- and sex-matched healthy subjects (cut off value = 0.401). Levels of anti-beta 2 GPI antibodies were linearly correlated with those of cardiolipin-reactive (aCL) antibodies. Eleven out of 22 patients (50%) had values of anti-prothrombin antibodies exceeding the cut-off value of 0.250. No relationship was found between the levels of anti-beta 2GPI and anti-prothrombin antibodies. Tests for antibodies against two natural inhibitors of blood coagulation, protein C and protein S, revealed elevated levels of anti-protein C IgG and anti-protein S IgG in 4 and 12 patients, respectively. A highly significant correlation between anti-protein C IgG and anti-protein S IgG values as well as between antibody titers against the two studied natural coagulation inhibitors and anti-prothrombin IgG was found. When comparing patients positive for aCL and presence or absence of a previous thrombotic episode (aCL+/T+ vs aCL+/T-), the positivity of anti-beta 2GPI IgG was found to be statistically associated with thrombosis. Conversely, among patients with previous thromboembolism with or without aCL (aCL+/T+ vs aCL-/T+ vs aCL-/T+) the positivity of anti-beta 2GPI IgG was strictly associated with the positivity of aCL, thus identifying the aPL antibody syndrome. These data demonstrate that anti beta 2GPI antibodies are a marker of "autoimmune" thrombosis. Anti-prothrombin antibodies are not a marker of thrombosis and are closely associated with antibodies to protein C and protein S. PMID- 8725713 TI - Species specificity of anti-beta 2 glycoprotein I autoantibodies and its relevance to anticardiolipin antibody quantitation. AB - Some patients suspected of having antiphospholipid antibody syndrome (APS) were found to be positive for anti-beta 2 glycoprotein I (beta 2GPI) antibodies despite negative results for antibodies to cardiolipin (ACA). Since the major source of beta 2GPI in the ACA assay is animal (usually bovine) serum, we studied the influence on ACA quantitation of the species specificity of anti-beta 2GPI antibodies from patients with various autoimmune disorders, mostly systemic lupus erythematosus and primary APS. Ninety-seven sera were selected based on IgG (n = 76) or IgM (n = 64) positivity by ELISA using gamma-irradiated plates coated with human or bovine purified beta 2GPI. A higher proportion of IgM (43.7%) than IgG (7.9%) reacted to human, but not bovine, beta 2GPI. Furthermore, from the samples reactive to both proteins, the ratio of antibody level against bovine to that against human beta 2GPI was 1.08 +/- 0.58 for IgG and 0.58 +/- 0.3 for IgM (p < 10(-5)). IgG and IgM ACA were detected in 78 and 40 sera, respectively; concordance between the two ELISAs for ACA and anti-beta 2GPI antibodies was 94% for IgG and 75% for IgM. Out of 28 IgM showing recognition restricted to human beta 2GPI, 21 were missed by the ACA assay, possibly because of lower concentrations of beta 2GPI in those patients' sera. The antibody reactivity pattern towards human and bovine beta 2GPI of individual sera showed no variation with time and was related to the relative antibody avidity for each protein. A murine anti-human beta 2GPI monoclonal antibody, 9G1, that cross-reacts with bovine beta 2GPI, competed to a large extent with the patients' anti-beta 2GPI antibody binding sites whatever isotype involved or protein recognized. Therefore, anti-beta 2GPI antibodies of IgM isotype display a marked preference for human compared to bovine beta 2GPI responsible for frequent inconsistencies in the ACA assay. PMID- 8725714 TI - Predicting daily maintenance dose of fluindione, an oral anticoagulant drug. AB - Due to large inter-individual variations, the dose of vitamin K antagonist required to target the desired hypocoagulability is hardly predictible for a given patient, and the time needed to reach therapeutic equilibrium may be excessively long. This work reports on a simple method for predicting the daily maintenance dose of fluindione after the third intake. In a first step, 37 patients were delivered 20 mg of fluindione once a day, at 6 p.m. for 3 consecutive days. On the morning of the 4th day an INR was performed. During the following days the dose was adjusted to target an INR between 2 and 3. There was a good correlation (r = 0.83, p < 0.001) between the INR performed on the morning of day 4 and the daily maintenance dose determined later by successive approximations. This allowed us to write a decisional algorithm to predict the effective maintenance dose of fluindione from the INR performed on day 4. The usefulness and the safety of this approach was tested in a second prospective study on 46 patients receiving fluindione according to the same initial scheme. The predicted dose was compared to the effective dose soon after having reached the equilibrium, then 30 and 90 days after. To within 5 mg (one quarter of a tablet), the predicted dose was the effective dose in 98%, 86% and 81% of the patients at the 3 times respectively. The mean time needed to reach the therapeutic equilibrium was reduced from 13 days in the first study to 6 days in the second study. No hemorrhagic complication occurred. Thus the strategy formerly developed to predict the daily maintenance dose of warfarin from the prothrombin time ratio or the thrombotest performed 3 days after starting the treatment may also be applied to fluindione and the INR measurement. PMID- 8725715 TI - The therapeutic range for heparin therapy: relationship between six activated partial thromboplastin time reagents and two heparin assays. AB - The activated partial thromboplastin time (APTT) is the most commonly used test for laboratory monitoring of unfractionated heparin therapy. Since there are differences between APTT reagents in respect of responsiveness to heparin the widely used therapeutic range of 1.5-2.5 (APTT ratios) may not be appropriate for all reagents. The aim of this study was to assess the relationship between 6 different APTT reagents using a manual technique, 2 of these reagents used in combination with a coagulometer, a heparin assay by protamine titration and a chromogenic anti-Xa assay. Samples from 42 patients treated with unfractionated heparin for thromboembolic disease were studied, 12 of whom were receiving warfarin therapy with International Normalised Ratios (INR) of > 1.3. For normal subjects, APTT results were highly dependent on the method used and statistically significant differences were noted. The ratio of patient to mean normal APTT was calculated for each APTT method. When 30 samples from heparinised patients (with INRs of < 1.3) were analysed manually, the APTT ranges equivalent to 0.2-0.4 u/ml heparin by protamine titration (by regression analysis) were 1.6-1.9 for Boehringer reagent (the least responsive) up to 2.2-2.9 for Instrumentation Laboratory reagent (the most responsive). The concentration of heparin associated on average with APTT ratios of 1.5-2.5 varied approximately twofold to threefold between reagents. PMID- 8725716 TI - Long-term persistence of biological activity following administration of Enoxaparin sodium (clexane) is due to sequestration of antithrombin-binding low molecular weight fragments--comparison with unfractionated heparin. AB - We have previously reported (Brieger D, Dawes J. Thromb Haemost 1994; 72: 275-80) that the prolonged anti-Xa amidolytic activity following intravenous administration of the low molecular weight heparin Enoxaparin sodium is mediated by small molecules derived from the injected drug, and an antithrombin binding penta/hexasaccharide can be detected in the circulation as late as 1 week after administration. To investigate the mechanism underlying this persistence we administered 125I-labelled fractions of Enoxaparin sodium and unfractionated 125I heparin to rabbits. Both 125I-heparin and the radiolabeled high molecular weight (> 6000 Da) Enoxaparin sodium were more effectively cleared from the circulation than the smaller components of LMW heparin. However, our data suggest that the circulating biologically active penta/hexasaccharide was not an unmodified component of the injected drug but was derived from a subpopulation of molecules of intermediate molecular weight (1800-6000 Da) which was retained in the tissues. Significant quantities of both Enoxaparin sodium and unfractionated heparin were retained in the internal organs. We propose that the sequestered subpopulations of Enoxaparin sodium and unfractionated heparin follow different catabolic routes. After administration of both unfractionated and LMW heparin additional antithrombin binding material was released into the circulation by a bolus dose of heparin. This material was not contained on circulating blood cells and was probably sequestered on the endothelium. PMID- 8725718 TI - Instrument effect on the activated protein C resistance plasma assay performed by a commercial kit. AB - In order to evaluate the influence of the coagulation instrument on the activated protein C (APC)-resistance plasma assay performed by a commercial kit, we tested 70 plasma samples on 4 different instruments during a simultaneous session run using a same lot of Coatest APC-resistance (Chromogenix). The results were analyzed employing three different modes of expression (aPTT prolongation in the presence of APC, APC-sensitivity ratio, normalized APC-sensitivity ratio) and three different diagnostic threshold values (below the control mean--2 standard deviations or the lowest control value or the 5th percentile of the control values). The inter-instruments variability in the mean values of the control individuals can be limited expressing the results as normalized-APC-sensitivity ratio (range 0.99-1.05). The overall diagnostic yield in thrombotic patients and their relatives depended mainly on the instrument employed and only in some cases on the mode of expression of the results and on the diagnostic threshold value. The sensitivity of the commercial assay on heterozygotes for factor V Leiden diagnosed by gene analysis was overall satisfactory (75-100%) but in some cases a lower diagnostic yield was noticed, depending on the type of instrument employed and/or the type of expression of the results and/or the diagnostic threshold values. Thus the instrument system adopted should be carefully considered in the interpretation of the results using the commercial kit. PMID- 8725717 TI - The effects of dermatan sulfate at submicrogram/ml concentrations on in vitro thrombin generation. AB - Dermatan sulfate is an antithrombotic glycosaminoglycan which has been shown to be effective in preventing deep venous thrombosis in general surgery patients when present at concentrations less than 1 microgram/ml. It has also been found to circulate physiologically in similar concentrations in pregnant women at term and in cord blood. We investigated the ability of dermatan sulfate added to plasma at 0.2, 0.5 and 1.0 microgram/ml to inhibit thrombin generation initiated by low concentrations of recombinant human tissue factor in defibrinated plasma. A dose dependent decrease in thrombin potential was demonstrated at therapeutically relevant concentrations of dermatan sulfate (0.5 and 1.0 microgram/ml) but there was no induction of a lag phase in thrombin generation. We were unable to demonstrate a significant effect on thrombin potential of dermatan sulfate at a concentration similar to that found in pregnancy plasma (0.2 microgram/ml). This indicates that either the dermatan sulfate concentration found in pregnancy plasma is not physiologically relevant or that our experimental system (which lacks platelets and fibrin) does not accurately reflect physiologic conditions. The effect on the thrombin potential was somewhat greater at the lowest concentration of tissue factor and amounted to a maximum inhibition of approximately 50% at 1 microgram/ml dermatan sulfate. A dose dependent increase in formation of thrombin-heparin cofactor II complexes and a decrease in thrombin-antithrombin complex formation with increasing dermatan sulfate concentration were observed at all dermatan sulfate concentrations. Prothrombin consumption was not changed by any dose of dermatan sulfate. We conclude that dermatan sulfate, at the concentrations tested, catalyses inhibition of free thrombin by heparin cofactor II but not efficiently enough to inhibit prothrombinase formation. PMID- 8725719 TI - Simple and rapid detection of factor V Leiden by allele-specific PCR amplification. AB - Resistance to activated protein C is the most common hereditary cause for thrombosis and significantly linked to factor V Leiden. In this study, primers were designed to identify the factor V mutation by allele-specific PCR amplification. 126 patients with thromboembolic events were analysed using this technique, PCR-RFLP and direct sequencing. The concordance between these techniques was 100%. In 27 patients a heterozygous factor VGln506 mutation was detected, whereas one patient with recurrent thromboembolism was homozygous for the point mutation. Due to its time- and cost-saving features allele-specific amplification should be considered for screening of factor VGln506. PMID- 8725720 TI - Role of the A+ helix in heparin binding to protein C inhibitor. AB - Interactions between proteins and heparin(-like) structures involve electrostatic forces and structural features. Based on charge distributions in the linear sequence of protein C inhibitor (PCI), two positively charged regions of PCI were proposed as possible candidates for this interaction. The first region, the A+ helix, is located at the N-terminus (residues 1-11), whereas the second region, the H helix, is positioned between residues 264 and 280 of PCI. Competition experiments with synthetic peptides based on the sequence of these regions demonstrated that the H helix has the highest affinity for heparin. In contrast to previous observations we found that the A+ helix peptide competed for the interaction of PCI with heparin, but its affinity was much lower than that of the H helix peptide. Recombinant PCI was also used to investigate the role of the A+ helix in heparin binding. Full-length (wild-type) rPCI as well as an A+ helix deletion mutant of PCI (rPCI-delta 2-11) were expressed in baby hamster kidney cells and both had normal inhibition activity with activated protein C and thrombin. The interaction of the recombinant PCIs with heparin was investigated and compared to plasma PCI. The A+ helix deletion mutant showed a decreased affinity for heparin in inhibition reactions with activated protein C and thrombin, but had similar association constants compared to wild-type rPCI. The synthetic A+ helix peptide competed with rPCI-delta w-11 for binding to heparin. This indicated that the interaction between PCI and heparin is fairly non specific and that the interaction is primarily based on electrostatic interactions. In summary, our data suggest that the H helix of PCI is the main heparin binding region of PCI, but the A+ helix increases the overall affinity for the PCI-heparin interaction by contributing a second positively charged region to the surface of PCI. PMID- 8725721 TI - Increased activation of the haemostatic system in men at high risk of fatal coronary heart disease. AB - The haemostatic system was examined in 2951 men aged 50 to 61 years, clinically free of cardiovascular disease, who were ranked according to a risk score for fatal coronary heart disease (CHD). Risk was judged from their serum cholesterol concentration, systolic blood pressure, body mass index and smoking habit. The status of the factor VII-tissue factor pathway was estimated from the plasma levels of factor VII coagulant activity, factor VII antigen and activated factor VII. Activation of factor IX was assessed from the plasma concentration of factor IX activation peptide. Activity within the common pathway was measured as the plasma concentrations of prothrombin fragment 1 + 2 and fibrinopeptide A. All 6 markers of haemostatic status were positively and statistically significantly associated with risk, providing further evidence for a hypercoagulable state in men at high risk for fatal CHD. Plasma fibrinogen and serum triglyceride concentrations were also graded positively with risk. PMID- 8725722 TI - Detection of circulating tissue factor and factor VII in a normal population. AB - The plasma tissue factor (TF) concentration was correlated to factor VII concentration (FVIIag) and factor VII activity (FVIIc) in 498 healthy volunteers ranging in age from 17 to 64 years. Immunoassays using monoclonal antibodies (mAbs) were developed for the determination of TF and FVIIag in plasma. The mAbs and the test systems were characterized. The mean value of the TF concentration was 172 +/- 135 pg/ml. TF showed no age- and gender-related differences. For the total population, FVIIc, determined by a clotting test, was 110 +/- 15% and the factor VIIag was 0.77 +/- 0.19 microgram/ml. FVII activity was significantly increased with age, whereas the concentration demonstrated no correlation to age in this population. FVII concentration is highly correlated with the activity as measured by clotting assay using rabbit thromboplastin. The ratio between FVIIc and FVIIag was not age-dependent, but demonstrated a significant difference between men and women. Between TF and FVII we could not detect a correlation. PMID- 8725723 TI - Fibrinogen and factor VIIag in healthy adolescents: the Floren-teen (Florence teenager) Study. AB - At least five studies based on more than twenty thousand healthy subjects indicated that fibrinogen is an independent risk factor for cardiovascular events; less clear-cut is the relation between factor VII and risk for arterial thrombotic disorders, which was demonstrated in two of the three studies investigating this association. However, no study has investigated the behaviour of fibrinogen and factor VII in an adolescent population. In a study of Preventive Medicine and Education Program, fibrinogen (clotting method) and factor VIIag (ELISA), in addition to other metabolic parameters, life-style and familial history, were investigated in 451 students (313 females and 138 males, age 15-17 years) from two high schools of Florence. Fibrinogen levels were significantly higher in women than in men, whereas factor VIIag levels did not significantly differ. Both fibrinogen and factor VIIag significantly correlated with total cholesterol (p < 0.05) while only fibrinogen correlated with body mass index (p < 0.01). Factor VIIag was significantly correlated with systolic blood pressure (p < 0.001). This study provides information on coagulation risk factors in a population of adolescents which may be of importance in planning coronary heart disease prevention programs. PMID- 8725724 TI - Fibrinogen promotes adhesion of monocytic to human mesothelioma cells. AB - Adhesion between monocytic and mesothelioma or pleural mesothelial cells influences stromal remodeling in pleural neoplasia. We found that cultured monocytic cells (U937) adhere to either human pleural mesothelioma (MS-1) or mesothelial (MeT5A) cells in vitro. 125I-fibrinogen bound specifically and saturably to either cell line, and specific fibrinogen binding increased upon stimulation of these cells with proinflammatory agents such as phorbol myristate (PMA), lipopolysaccharide (LPS) or tumor necrosis factor (TNF-alpha). We purified the fibrinogen receptor protein from a membrane fraction of MS-1 cells and identified it by immunoprecipitation as intercellular adhesion molecule (ICAM-1). Anti-ICAM-1 antibody or antisense oligonucleotides inhibited fibrinogen-mediated cell adhesion and binding of 125I-fibrinogen to mesothelioma or mesothelial cells. Cultured monocytic cells adhere to either mesothelioma or mesothelial cells, and the interaction is promoted by fibrinogen binding ICAM-1 at the cell surface. ICAM-1 is expressed by mesothelioma cells and CD 11b by macrophages in the fibrinous mesothelioma tumor stroma. The data suggest a common mechanism by which monocytic cells could adhere to either malignant mesothelioma cells or the mesothelial surface in pleural neoplasia. PMID- 8725725 TI - Macrophages and neutrophils infiltrating into the liver are responsible for tissue factor expression in a rabbit model of acute obstructive cholangitis. AB - Acute obstructive cholangitis (AOC) is one of the most fatal outcomes in sepsis, and frequently complicates disseminated intravascular coagulation (DIC). Recently we found that the plasma tissue factor (TF) level increased and changed in parallel with plasma markers of DIC in patients with AOC. To elucidate the role of TF in the pathogenesis of coagulopathy in AOC, we investigated the plasma levels of TF and its localization by immunohistochemical staining in rabbit models of AOC. Plasma TF activity significantly increased 3 h after the insult (0.63 +/- 0.1?9 U/ml; p < 0.01) compared with that beforehand (0.05 +/- 0.02 U/ml), then reached a maximum level at 6 h (0.94 +/- 0.16 U/ml). The fluctuations in plasma TF activity correlated with those of the coagulation parameters including platelet count, fibrinogen, prothrombin time, and antithrombin III activity. Immunohistochemically, enhanced expression of TF was mainly detected in macrophages and neutrophils that had infiltrated into the liver sinusoids and around the bile duct, but not in the sinusoidal endothelial cells. A double immunofluorescence study revealed the concomitant presence of TF and fibrin at sites where macrophages and neutrophils had conglomerated. However, we could not detect an apparent change in TF expression in the lung or kidney. These data suggest that macrophages and neutrophils infiltrating into the liver sinusoids and around the bile duct play a pivotal role in TF expression, leading to coagulopathy in the acute phase of obstructive cholangitis in rabbits. PMID- 8725726 TI - Factor Xa enhances the binding of tissue factor pathway inhibitor to acidic phospholipids. AB - In the present study, the interaction between tissue factor pathway inhibitor (TFPI) and phospholipids has been characterized using a microtitre plate assay. TFPI was shown to bind calcium-independently to an acidic phospholipid surface composed of phosphatidylserine, but not a surface composed of the neutral phosphatidylcholine. The interaction was demonstrated to be dependent on the presence of the TFPI C-terminus. The presence of heparin (1 U/ml, unfractionated) was able to significantly reduce the binding of TFPI to phospholipid. The interaction of TFPI with phosphatidylserine was significantly decreased in the presence of calcium, but this was counteracted, and even enhanced, following complex formation of TFPI with factor Xa prior to incubation with the phospholipid surface. Moreover, a TFPI variant, not containing the third Kunitz domain and the C-terminus, was unable to bind to phospholipid. However, following the formation of a TFPI/factor Xa-complex this TFPI variant was capable of interacting with the phospholipid surface. This indicates that the role of factor Xa as a TFPI cofactor, at least in part, is to mediate the binding of TFPI to the phospholipid surface. PMID- 8725727 TI - Production of tissue-type plasminogen activator (t-PA) and type-1 plasminogen activator inhibitor (PAI-1) in mildly cirrhotic rat liver. AB - We have studied the production of tissue-type plasminogen activator (t-PA) and type-1 plasminogen activator inhibitor (PAI-1) in liver of normal rats and in rats with mild cirrhosis induced by carbon tetrachloride inhalation, to demonstrate the production of these fibrinolytic components and their pathophysiologic role in the liver in vivo. Immunohistochemical study of paraffin embedded liver sections and fibrin autography of frozen sections showed that the normal rat liver produces very little t-PA or PAI-1. On the contrary, striking t PA activity and both t-PA and PAI-1 antigens were observed in the cirrhotic liver. Both t-PA and PAI-1 in plasma were also markedly increased in the cirrhotic rats. Because the hepatocyte can internalize t-PA or PA/PAI-1 complexes from circulation, Northern blot analysis of the total liver RNA was performed to demonstrate the endogenous synthesis of t-PA and PAI in the liver. Although the normal liver hardly expresses either t-PA or PAI-1 mRNA, striking t-PA and PAI-1 mRNA expression was observed in the liver of rats with mild cirrhosis. These data demonstrate that t-PA and PAI-1 production is strongly upregulated in the liver in rats with mild cirrhosis. These fibrinolytic components, whose production is closely associated with liver failure, may play important roles in the regulation of hepatocyte proliferation and liver regeneration in vivo. PMID- 8725729 TI - Comparative effects of staphylokinase and alteplase in rabbit bleeding time models. AB - BACKGROUND: The pathogenesis of bleeding associated with thrombolytic therapy remains largely unknown, although spontaneous bleeding appears to correlate with bleeding time prolongation. Here, the comparative effects on cuticle bleeding times (CBT) and ear puncture bleeding times (EBT) of recombinant staphylokinase (Sak) and alteplase (recombinant tissue-type plasminogen activator, rt-PA) at equivalent doses, alone and in combination with aspirin and heparin, were studied in rabbits. METHODS AND RESULTS: Groups of 4 to 9 rabbits were allocated to one of the 8 following intravenous infusions: saline; aspirin 15 mg/kg and heparin - 100 IU/kg bolus and 10 IU/kg infusion over one hour; 1.5 mg/kg rt-PA; 1.5 mg/kg rt-PA plus aspirin and heparin; 4.5 mg/kg rt-PA; 0.5 mg/kg Sak; 0.5 mg/kg Sak plus aspirin and heparin and 1.5 mg/kg Sak. Bleeding times were determined 30 and 15 min before and 5, 15, 30 and 60 min after the administration over one min of saline, rt-PA or Sak, by simultaneously severing a nail cuticle (CBT) and by puncturing the ear (EBT). Bleeding times were unaffected by saline and by both doses of Sak in monotherapy. Heparin-aspirin and low dose rt-PA significantly lengthened EBT but not CBT. Both CBT and EBT were significantly prolonged (to a mean of > 4 times pretreatment at 5 min) after high-dose rt-PA and after the combined administration of heparin and aspirin with either Sak or tr-PA. rt-PA provoked significantly longer bleeding than Sak in the CBT (p = 0.001; mean estimated difference = 23 min), but not in the EBT. rt-PA but not Sak degraded plasma fibrinogen dose-dependently. CBT correlated inversely with fibrinogen (r= 0.66, p=0.001) but EBT did not. CONCLUSION: At equivalent doses Sak displays a significantly higher fibrin specificity and prolongs bleeding time less than rt PA, particularly in the nail cuticle bleeding time model in which larger vessels are injured that require fibrinogen for hemostasis. PMID- 8725728 TI - Evaluation of a low molecular weight modulator of human plasminogen activator inhibitor-1 activity. AB - A critical component in the regulation of thrombus formation and clearance is the balance between tissue plasminogen activator (tPA) and plasminogen activator inhibitor type-1 (PAI-1). An increase in the plasma concentration of PAI-1 has been proposed as a risk factor in thrombotic disease. Inhibition of PAI-1 activity may have utility in the treatment of thromboembolic disease. We report here the evaluation of three diketopiperazine-based low molecular weight inhibitors of PAI-1 activity (XR334, XR1853 and XR5082). In vitro these compounds reversed the inhibitory effects of PAI-1 against both tPA and urokinase (UK) (IC50: 5 to 80 muM). In contrast, other serpin-serine protease interactions, including alpha 1-antitrypsin-trypsin, alpha 2-antiplasmin- plasmin and antithrombin-thrombin, were not affected, neither did these inhibitors affect global tests of haemostasis. In the light of this promising in vitro profile these compounds were evaluated in a standard radioisotopic assay of clot lysis in whole rat blood following intravenous administration. In this assay these compounds dose-dependently enhanced fibrinolysis ex vivo. After intravenous bolus administration XR334, XR1853 and XR5082 at 5 mg/kg increased clot lysis by 32.0 +/- 5.1% SEM (n = 25, p < 0.01), 36.7 +/- 3.5% SEM (n = 36, p < 0.01) and 60.0 +/ 2.8% SEM (n = 17, p < 0.01) respectively compared to vehicle. Intravenous infusion of these compounds (1 mg/kg/min for 20 min) significantly prolonged (approximately twofold) the time to blood vessel occlusion in the rat electrically-stimulated carotid artery thrombosis model. Thus, these low molecular weight inhibitors of PAI-1 activity enhanced fibrinolysis ex vivo and protected against thrombus formation in the rat. PMID- 8725730 TI - An experimental model of intracranial hemorrhage during thrombolytic therapy with t-PA. AB - Multiple clinical trials have proven that thrombolytic therapy is an effective treatment for acute myocardial infarction. Spontaneous intracranial hemorrhage (ICH) occurs in a small percentage of patients as a result of the treatment. The etiology of the ICH is unknown and there is currently no established experimental model for this side effect. A model of ICH during thrombolytic therapy has been developed using spontaneously hypertensive rats (SHR). The SHR were made susceptible to ICH during thrombolytic therapy by bilateral ligation of the external jugular veins. This procedure produced asymptomatic hemorrhagic lesions in the brains of the animals in the hours preceding the administration of t PA/heparin. The incidence of ICH following the administration of test substances was assessed by histological examination and by measuring the red blood cell count in a sample of cerebrospinal fluid taken from the atlanto-occipital space. t-PA administration produced a low frequency of ICH in this model. The incidence and severity of ICH were dramatically increased, and significant mortality at 24h was observed, by combining heparin were administered sequentially rather than simultaneously. Furthermore, ICHs were observed whether the t-PA dose was administered over 4 h, 1 h, or as a double bolus 30 min apart. The potentiation of ICH by heparin was dose dependent and proportional to the prolongation of the aPTT. Although the precise mechanism of ICH during thrombolytic therapy is unknown, many similarities exist between the observations made in this model and in the human clinical experience. PMID- 8725731 TI - Reduced effect of aspirin on thrombus formation at high shear and disturbed laminar blood flow. AB - Aspirin is the most commonly used antithrombotic drug in primary and secondary prophylaxis against cardio- and cerebrovascular disease. In previous studies from our laboratory it was demonstrated that the effect of aspirin on collagen-induced thrombus formation in a parallel- plate perfusion device with laminar blood flow is shear rate dependent. Although aspirin did not affect collagen-induced thrombus formation at 650 s-1 (medium sized arteries), a significant inhibition of thrombus formation by approximately 38% at 2,600 s-1 (moderately stenoses in medium sized arteries) was observed. At present we have extended these studies to thrombus formation at the apex of eccentric stenoses in a parallel-plate perfusion chamber device. The stenoses reduced the cross-sectional area of the blood flow channel of the perfusion chambers by 60 or 80%, introducing disturbed laminar flow and apex wall shear rates of 2,600 and 10,500 s-1, respectively. The corresponding wall shear stresses were 80 and 315 dynes/cm2, respectively. Aspirin reduced the platelet thrombus volume at the 60% stenosis by 45% (p < 0.03), and the fibrin deposition by 70% (p < 0.004). However, none of these parameters were affected by aspirin at the 80% stenosis. These observations may at least partly explain why aspirin has a limited clinical effect in preventing arterial thrombus formation in atherosclerotic vessels at high shear and disturbed blood flow. In contrast, thrombus formation in blood from one patient with Glanzmann's thrombasthenia and two patients with von Willebrand disease subtype 2M was almost abolished at this blood flow condition. Thus, blocking the function of either von Willebrand factor or glycoprotein IIb/IIIa may represent better antithrombotic approaches for such critical events than blocking the prostaglandin metabolism by aspirin. The lack of effect of aspirin on thrombus formation at the 80% stenosis may reflect shear-induced platelet activation at the stenosis inlet region, since shear-induced platelet aggregation in rotational viscometers is not affected by aspirin at shear stresses exceeding 100 dynes/cm2. PMID- 8725732 TI - Enhanced platelet aggregability under high shear stress after treadmill exercise in patients with effort angina. AB - Platelet activation induced by shear forces occurring in a stenosed coronary artery is one of the mechanisms of coronary thrombosis. We evaluated the shear induced platelet aggregation (SIPA) dynamics in patients with effort angina during treadmill exercise. SIPA was measured by a rotational cone-plate aggregometer. SIPA was markedly increased by exercise from 71.2 +/- 8.9% to 81.9 +/- 7.6% (p < 0.01) in the patient group. Although epinephrine concentrations were elevated, its rate of increase was not correlated with that of SIPA. Yohimbine partially inhibited the exercise-induced increase in SIPA. In contrast, a significant correlation between the changing rate of plasma von Willebrand factor (vWF) larger multimers and that of SIPA (r = 0.74, p < 0.05) was observed. Exercise-augmented SIPA is probably dependent on an increase in vWF larger multimers rather than platelet alpha2-receptor activation. Prevention of the interaction between vWF and its platelet receptors may play some role in decreasing the risk of coronary thrombosis during exercise. PMID- 8725733 TI - Development of large platelet aggregates from small aggregates as determined by laser-light scattering: effects of aggregant concentration and antiplatelet medication. AB - Particle-counting methods that employ light scattering (LS) quantify changes in the number of platelet aggregates of different sizes after the application of an aggregating stimulus. Using the LS method, we studied the effects of aggregant concentration, aspirin administration, and ticlopidine administration on aggregate formation and compared the results with those obtained using the conventional optical density (OD) method. Subjects were 47 controls, 31 patients treated with aspirin (330 mg/day), and 37 patients treated with ticlopidine (200 mg/day). Platelet aggregation after stimulation by 0.5, 1.0 or 5.0 muM ADP, or 0.5, 1.0 or 2.0 micrograms/ml collagen was determined using both methods. Using the LS method, small (9-25 micrograms), medium (25-50 micrograms), and large (50 70 micrograms) aggregates were counted. In patients untreated with antiplatelet medication, greater concentrations of ADP or collagen generated larger aggregates. Generation of small and medium-sized aggregates showed a significant positive correlation with OD levels after stimulation with 0.5 or 1.0 muM ADP, or 0.5 or 1.0 micrograms/ml collagen. In patients treated with aspirin, the development of small aggregates into large aggregates was inhibited. Thus, the number of small aggregates increased. Inhibition induced by aspirin was more effective against aggregation after stimulation with collagen than with ADP. In patients treated with ticlopidine, small and medium-sized aggregate formation was inhibited after stimulation with low concentrations of ADP or collagen, but was promoted after stimulation with high aggregant concentrations. The capability of the LS method to quantify different sizes of aggregates after stimulation with low concentration agonists may facilitate investigation of the aggregation process, and of how this process is affected by antiplatelet agents. PMID- 8725734 TI - Aprotinin has no effect on platelet activation and adhesion during cardiopulmonary bypass. AB - Aprotinin reduces blood loss following cardiopulmonary bypass operations (CPB) by the prevention of hyperfibrinolysis. Its influence on circulating platelets is uncertain. In this prospective trial we investigated activation, adhesion, and aggregation receptors on the platelet surface in 20 patients who underwent elective coronary artery bypass grafting. These patients were randomly assigned to receive either a high dose of aprotinin or placebo. Flow cytometry was performed to determine platelet activation [P-selectin, glycoprotein (GP) 53], adhesive (GP Ib), and aggregatory (GP IIb-IIIa) receptors on circulating platelets, before, during, and after CPB. Aprotinin had neither a significant effect on platelet activation nor on adhesive and aggregatory receptors. Plasma levels of D-dimers were measured before and after CPB to assess fibrinolytic activity. D-dimers following CPB and chest tube drainage were significantly less in the aprotinin group. We conclude that aprotinin reduces blood loss by its effect on fibrinolysis but has no direct influence on platelet function. PMID- 8725735 TI - Ablation characteristics of arterial vessel walls irradiated with the pulsed dye laser--an analysis of variance. AB - BACKGROUND AND OBJECTIVE: Since most clinical laser angioplasties require the use of over-the-wire delivery systems, we studied the effects of pulsed dye laser energy (504 nm, 1.4 microseconds on arterial vessel walls in combination with a multifiber catheter system. MATERIAL AND METHODS: Postmortem arterial segments (n = 368) were exposed under blood or saline. Laser pulses (n = 100-800) were transmitted via 9F-multifiber-catheters, at energy densities of 3-16 J/cm2. Ablation characteristics revealed by histologic examination and morphometry were analyzed by multiple analysis of variance. RESULTS: Ablation occurred more frequently in saline compared to blood. Below an energy density of 10 J/cm2 ablation occurred in saline only. Specimens irradiated under blood showed only thermal changes at 10 J/cm2. In saline, 92% of normal, 88% of fibro-fatty and 60% of calcified tissue showed ablation at 13 J/cm2. The average ablation threshold in saline was about 3-4 J/cm2 per pulse for normal tissue, 5 J/cm2 for fatty plaques, and 8-9 J/cm2 for calcified plaques. In blood, the average ablation thresholds did not differ significantly between the different stages of arteriosclerosis (12 J/cm2 for normal tissue, 11 J/cm2 for fatty plaque, and 10 J/cm2 for calcified tissue). Carbonization and vacuolization were seen regularly at energy levels > or = 13.4 J/cm2. CONCLUSIONS: Selective ablation of arteriosclerotic tissue with the pulsed dye laser could not be found. Further investigation is needed before an effective ablation of arteriosclerotic arterial tissue can be expected. PMID- 8725736 TI - Evidence for cultured human vascular smooth muscle cell heterogeneity: isolation of clonal cells and study of their growth characteristics. AB - The monoclonal theory of atherosclerosis postulates that the initial vascular smooth muscle cell (VSMC) proliferative event involves the expansion of a single cell or a sub-population of cells thus implying differences in the replicative potential of VSMC. Using the technique of limited dilution, VSMC clones derived from animal tissues have been previously isolated and shown to be morphologically heterogeneous. However, the same technique applied to human VSMC (HVSMC) has been unsuccessful, possibly because HVSMC do not grow when plated at very low densities. In this report, the anchorage-independent growth factor-BB (PDGF-BB) and to lesser extent PDGF-AB and basic fibroblast growth factor (bFGF) induced colony formation. This assay provided a tool for the isolation of HVSMC clones. In terms of their growth characteristics and responsiveness to several growth factors, isolated HVSMC clones and the original parental cell population exhibited marked heterogeneity. PMID- 8725737 TI - Antiphospholipid antibodies, recurrent thromboembolism, and intensity of warfarin anticoagulation. PMID- 8725738 TI - Factor V Leiden and sickle cell disease. PMID- 8725739 TI - Very low incidence of Arg506-->Gin mutation in the factor V gene among the Amazonian Indians and the Brazilian black population. PMID- 8725740 TI - Absence of length polymorphism within the 31 tandem repeats of the B domain of factor V. PMID- 8725741 TI - Aspirin, vitamin E and heart attack prevention. PMID- 8725742 TI - Clinical similarities in siblings with schizophrenia. AB - Three symptom groups, identified by factor analysis of schizophrenic symptoms, together with other clinical variables, were compared among 80 sibships (169 individuals) containing two or more members affected with schizophrenia. The three factors, which were labelled negative symptom disorganization, and reality distortion, all showed a moderate but significant degree of correlation between siblings. Age at onset was also significantly correlated. Such a familial pattern of clinical heterogeneity suggests underlying common familial aetiologies that influence the clinical form of the disorder. Whether these are genetic or environmental requires further investigation. This finding confers some external validation on the three factor model. It may be feasible to develop familial symptom patterns as the basis for an a priori approach to linkage heterogeneity. PMID- 8725743 TI - Use of linkage disequilibrium approaches to map genes for bipolar disorder in the Costa Rican population. AB - Linkage disequilibrium (LD) analysis provides a powerful means for screening the genome to map the location of disease genes, such as those for bipolar disorder (BP). As described in this paper, the population of the Central Valley of Costa Rica, which is descended from a small number of founders, should be suitable for LD mapping; this assertion is supported by reconstruction of extended haplotypes shared by distantly related individuals in this population suffering low frequency hearing loss (LFHL1), which has previously been mapped by linkage analysis. A sampling strategy is described for applying LD methods to map genes for BP, and clinical and demographic characteristics of an initially collected sample are discussed. This sample will provide a complement to a previously collected set of Costa Rican BP families which is under investigation using standard linkage analysis. PMID- 8725744 TI - An approach to investigating linkage for bipolar disorder using large Costa Rican pedigrees. AB - Despite the evidence that major gene effects exist for bipolar disorder (BP), efforts to map BP loci have so far been unsuccessful. A strategy for mapping BP loci is described, focused on investigation of large pedigrees from a genetically homogenous population, that of Costa Rica. This approach is based on the use of a conservative definition of the BP phenotype in preparation for whole genome screening with polymorphic markers. Linkage simulation analyses are utilized to indicate the probability of detecting evidence suggestive of linkage, using these pedigrees. These analyses are performed under a series of single locus models, ranging from recessive to nearly dominant, utilizing both lod score and affected pedigree member analyses. Additional calculations demonstrate that with any of the models employed, most of the information for linkage derives from affected rather than unaffected individuals. PMID- 8725746 TI - Genetic heterogeneity in catatonic schizophrenia: a family study. AB - In family study concentrating on 139 probands with chronic DSM-III-R schizophrenia, catatonic type, 83 probands (41 women, 42 men) met the criteria for periodic catatonia and 56 probands (14 women, 42 men) for systematic catatonia according to the Leonhard classification. The reliability and stability of this subclassification were tested by 2 experienced psychiatrists working independently of each other. Both diagnosticians were kept blind as to the probands' family history. The 139 probands had a total of 543 first-degree relatives. Only those hospitalized for schizophrenia were allocated to the group of afflicted family members. Diagnostic reliability was kappa statistic 0.93 and diagnostic stability during catamnesis reached 97% and kappa of 0.93. Life-table analyses revealed that the age-corrected risks were significantly different in periodic and systematic catatonia. In systematic catatonia mothers had a risk of 6.8%, fathers 2%, and randomly selected sibs 3%. IN periodic catatonia an excess of homologous psychoses was apparent: There was a risk of 33.7% for mothers, 15.4% for fathers, and 24.4% for sibs. The quota of afflicted parents (33 of 161) was higher than that of sibs (26 of 162). In periodic catatonia, 59% of the families were multiple afflicted with pronounced unilineal vertical transmission. In 10% of the families 3 successive generations suffered from the disease and were treated in hospital. The results of the study led to the following hypotheses: Periodic and systematic catatonia are valid subgroups of DSM-III-R schizophrenia. In systematic catatonia heritability is very low. Periodic catatonia is a familial disorder. Homogeneity of familial psychoses and unilineal vertical transmission with anticipation are consistent with a major gene effect. Periodic catatonia seems to be a promising candidate for molecular genetic evaluation. PMID- 8725745 TI - Polygenic inheritance of Tourette syndrome, stuttering, attention deficit hyperactivity, conduct, and oppositional defiant disorder: the additive and subtractive effect of the three dopaminergic genes--DRD2, D beta H, and DAT1. AB - Polymorphisms of three different dopaminergic genes, dopamine D2 receptor (DRD2), dopamine beta-hydroxylase (D beta H), and dopamine transporter (DAT1), were examined in Tourette syndrome (TS) probands, their relatives, and controls. Each gene individually showed a significant correlation with various behavioral variables in these subjects. The additive and substractive effects of the three genes were examined by genotyping all three genes in the same set of subjects. For 9 of 20 TS associated comorbid behaviors there was a significant linear association between the degree of loading for markers of three genes and the mean behavior scores. The behavior variables showing the significant associations were, in order attention deficit hyperactivity disorder (ADHD), stuttering oppositional defiant, tics, conduct, obsessive-compulsive, mania, alcohol abuse and general anxiety-behaviors that constitute the most overt clinical aspects of TS. For 16 of the 20 behavior scores there was a linear progressive decrease in the mean score with progressively lesser loading for the three gene markers. These results suggest that TS, ADHD, stuttering oppositional defiant and conduct disorder, and other behaviors associated with TS, are polygenic, due in part to these three dopaminergic genes, and that the genetics of other polygenic psychiatric disorders may be deciphered using this technique. PMID- 8725747 TI - No evidence for a major gene effect of the dopamine D4 receptor gene in the susceptibility to Gilles de la Tourette syndrome in five Canadian families. AB - Gilles de la Tourette Syndrome (TS) is neuropsychiatric disorder characterized by both motor and vocal tics affecting approximately 1/10,000 females and 1/2000 males. Because of the success of neuroleptics and other agents interacting with the dopaminergic system in the suppression of tics, a defect in the dopamine system has been hypothesized in the etiology of TS. In this paper we test the hypothesis that the dopamine D4 receptor (DRD4) is linked to the genetic susceptibility to TS in five families. We tested three polymorphisms in the DRD4 gene and a polymorphism in the closely linked locus, tyrosine hydroxylase (TH). We found no evidence for linkage of DRD4 or TH to TS using an autosomal dominant model with reduced penetrance or using non-parametric methods. The presence of a mutation that results in a truncated non-functional D4 receptor protein was also tested for, but was not observed in these families. PMID- 8725748 TI - Allelic association but only weak evidence for linkage to the apolipoprotein E locus in late-onset Swedish Alzheimer families. AB - An association between the epsilon 4 allele of the apolipoprotein E gene (APOE) and late-onset Alzheimer's disease (AD) was recently demonstrated. In order to confirm the association and to gauge the ability of standard genetic linkage methods to identify susceptibility genes, we investigated 15 Swedish late-onset Ad families. We found an association of familial AD to the APOE epsilon 4 allele (P = 0.01) but no indication of linkage to the APOE region using 2-point linkage analysis, and only weak evidence using the affected pedigree-member (APM) method. Our results confirm an APOE epsilon 4 association with late-onset familial AD and indicate that susceptibility genes can easily be missed when using standard lod score and APM genetic linkage analysis. PMID- 8725749 TI - The Meckel syndrome: report of two Japanese sibs and a review of literature. AB - Two Japanese sibs with the Meckel syndrome are reported. Both babies showed the classical triad of this conditions: occipital encephalocele, cystic kidneys, and postaxial polydactyly of all four limbs. The diagnostic criteria and differential diagnosis were reviewed. PMID- 8725750 TI - Autism and genetics: high incidence of specific genetic syndromes in 21 autistic adolescents and adults living in two residential homes in Belgium. PMID- 8725751 TI - Sacrospinous colpopexy in the management of uterovaginal prolapse. AB - OBJECTIVE: To investigate the morbidity and results of vaginal hysterectomy with concomitant sacrospinous colpopexy for uterovaginal prolapse. STUDY DESIGN: An observational study was carried out from June 17, 1986, to September 22, 1992. Patients were selected if it was thought that the cardinal-uterosacral ligaments could not be relied upon for vaginal vault support. RESULT: During the study period, 265 vaginal hysterectomies were performed. Forty-five (17%) were with concomitant sacrospinous colpopexy. The mean patient age was 54 years. There was one incidental cystotomy during hysterectomy, and two patients required transfusion. Postoperatively, eight patients were treated for soft tissue infection, one developed new-onset urinary incontinence, and no apparent nerve injuries were diagnosed. The mean day of discharge was 4.4. Six patients were lost to follow-up after the early postoperative period. The mean follow-up for the remaining patients was 29 months (12-66). One patient required subsequent vaginal repair for recurrent cystocele and enterocele. Four patients had persistent stress urinary incontinence. CONCLUSION: Sacrospinous colpopexy at the time of vaginal hysterectomy is reasonably safe and effective for reestablishing upper vaginal support. PMID- 8725752 TI - Laparoscopic surgical management of ovarian cysts assisted by simultaneous transvaginal ultrasonography. AB - OBJECTIVE: To assess the use of transvaginal ultrasound guidance during operative laparoscopy. STUDY DESIGN: Five of 98 consecutive cases of operative laparoscopy for benign-appearing ovarian cysts were complicated by extensive intrapelvic adhesions and a distorted anatomy. Following prolonged dissection, simultaneous transvaginal ultrasonography was used in these cases. RESULTS: In all cases we were able to ultrasonographically image and locate the ovarian cyst during the laparoscopic procedure. In four cases this was a significant aid in expediting the procedure and avoiding damage to adjacent pelvic structures. In one case, identification of the ovarian cyst facilitated the decision to resort to laparotomy. CONCLUSION: In selected cases, laparoscopic surgeons should be aware of the feasibility and possible benefits of the use of simultaneous intraoperative transvaginal ultrasonography. PMID- 8725753 TI - Use of optical catheters for diagnostic office laparoscopy. AB - OBJECTIVE: To evaluate several small optical catheters (1.8-mm outer diameter) developed to allow access to the pelvic cavity for laparoscopic diagnosis in the operating room and in an office setting. STUDY DESIGN: Thirty-six procedures are reported, 20 performed in the operating room so that proper placement could be monitored and ensured, and the findings were compared with those obtained with a standard, 10-mm laparoscope. The remaining 16 procedures were performed in an office setting. The patients were women who were candidates for second-look laparoscopy, had pelvic pain and no physical findings, or were evaluated for possible tubal anastomosis. CONCLUSION: The small, rigid catheter, with its excellent optics, provided a clear view of the pelvis without the need for general anesthesia and associated costs. The procedure was performed easily and successfully with a local anesthetic and with minimal discomfort for the patient. PMID- 8725754 TI - Single vs. multiple semen specimens in screening for male infertility factors. A comparison. AB - OBJECTIVE: To reevaluate the number of semen analyses necessary to establish whether further male infertility testing is necessary. STUDY DESIGN: The results of three consecutive semen analyses for infertility evaluations were retrospectively reviewed. A male factor was defined by an abnormal semen analysis if either the first specimen of three (single-sample screening) or two of the three specimens (multiple-sample screening) met World Health Organization criteria. Males considered abnormal by multiple-sample screening underwent sophisticated andrologic evaluation. RESULTS: A single-sample conventional semen analysis obtained from 209 males demonstrated a diagnostic accuracy of 10.4% false negatives and a sensitivity of 89.6% when compared to that of multiple sample analysis. Andrologic evaluation of abnormal males by multiple-sample screening confirmed that 9 of the 11 men with normal first specimens were abnormal; all others were confirmed as abnormal. CONCLUSION: Analysis of multiple semen specimens provides a reliable screen in the evaluation of male factor infertility when the goal is to minimize the false negative rate of screening tests. PMID- 8725755 TI - Uterine myomas and smoking. Results from an Italian study. AB - OBJECTIVE: To evaluate the relationship between smoking and uterine myomas requiring surgery. STUDY DESIGN: We conducted a case-control study in Milan between 1986 and 1992. Cases were 476 patients under 55 years of age with histologically confirmed myomas. Controls were 1,283 women admitted to the hospital for a spectrum of acute, other-than-gynecologic, hormonal or neoplastic conditions (30% trauma, 25% nontraumatic orthopedic conditions, 25% surgical, 20% other miscellaneous). RESULTS: Cases were less frequently current smokers (22%) than controls (32%). In comparison with never smokers, the multivariate relative risk (RR) for myomas was 0.5 (95% confidence interval [CI], 0.4-0.7) in smokers. Ex-smokers were 10% of cases versus 7% of controls (RR 1.2, 95% CI 0.9-1.8). No clear trend in risk was observed with the number of cigarettes smoked per day or duration of smoking and risk of fibroids. The estimated RRs were largely consistent when separate analyses were performed in strata of age and other selected covariates, including indices of body weight. CONCLUSION: Current smoking seems to reduce the risk of myomas. PMID- 8725756 TI - Suppression and flare regimens of gonadotropin-releasing hormone agonist. Use in women with different basal gonadotropin values in an in vitro fertilization program. AB - OBJECTIVE: To evaluate the suppression and flare regimens of gonadotropin releasing hormone agonist (GnRH-a) in ovarian hyperstimulation in women with variable basal gonadotropin values in an in vitro fertilization (IVF) program. STUDY DESIGN: A retrospective study comparing the initiation of GnRH-a in the midluteal phase of the preceding cycle (suppression protocol) and follicular phase of the stimulated cycle (flare protocol) in women with basal follicle stimulating hormone (FSH) values < 15 mIU/mL and > or = 15 mIU/mL. RESULTS: The pregnancy rate per initiated cycle and implantation rate for women with basal FSH levels > or = 15 mIU/mL were 20.4% and 9.8% in flare GnRH-a cycles and 11.7% and 3.5%, respectively, in suppression GnRH-a cycles. Comparing the percent differences in clinical pregnancy and implantation rates between both protocols for women with different basal FSH values, pregnancy outcome was significantly greater in the flare protocols in women with values > or = 15 mIU/mL (P < .001). Individualization of the stimulation protocol by retrospective sorting of women undergoing IVF with respect to their basal gonadotropin levels significantly improved clinical pregnancy (P < .05) and implantation rates (P < .05) and reduced the cancellation rate (P < .05). CONCLUSION: The flare regimen with GnRH a is a useful alternative for controlled ovarian hyperstimulation in women with elevated basal FSH values (> or = 15 mIU/mL) undergoing IVF. PMID- 8725757 TI - Predicting pregnancy outcome from the degree of maternal serum alpha-fetoprotein elevation. AB - OBJECTIVE: To determine if a correlation exists between the level of maternal serum alpha-fetoprotein (MSAFP) elevation and the rate of adverse pregnancy outcome, to examine the timing of pregnancies ending in fetal or neonatal death, and to develop a protocol for antepartum surveillance in an effort to prevent these adverse outcomes. STUDY DESIGN: Singleton pregnancies with a single second trimester elevated MSAFP > or = 2.0 multiples of the median (MoM) were eligible if a targeted ultrasound evaluation (< 24 weeks) was in agreement with the dates and no fetoplacental anomaly was detected. Three groups were established based on the second-trimester MSAFP elevation: 2.0-2.49, 2.5-2.99 and > or = 3.0 MoM. RESULTS: Among the 383 patients enrolled, delivery data were available on 333 infants. Stratified by MSAFP elevations of 2.0-2.49, 2.5-2.99 and > or = 3.0 MoM, the rates of adverse pregnancy outcome were: (1) preterm birth: 14.3%, 15.6%, 20.3%; (2) small for gestational age at birth: 7.4%, 11.1%, 22.2%; and (3) perinatal deaths (neonatal and fetal): 2.6%, 3.3%, 5.6%. Seven pregnancy losses (three neonatal and four fetal deaths) occurred prior to 28 weeks. Of these seven, six fetuses exhibited intrauterine growth retardation by 23-26 weeks' gestation, and five of six were associated with MSAFP levels > or = 2.5 MoM. Four losses (two neonatal and two fetal deaths) occurred after 28 weeks. Of these, three involved structurally normal infants with normal growth who died after 34 weeks. All three of these pregnancies exhibited MSAFP elevations < 2.5 MoM. CONCLUSION: In pregnancies with an unexplained elevated second-trimester MSAFP, the rate of adverse pregnancy outcomes is increased with higher elevations. Any proposed program to improve pregnancy outcome in patients with unexplained MSAFP elevations must include efforts aimed at preventing preterm delivery, repeat ultrasound at 24-26 weeks to rule out early-onset intrauterine growth retardation in pregnancies with elevations > or = 2.5 MoM and fetal biophysical monitoring, even in normally grown fetuses, instituted at 32 weeks to detect fetuses at risk for intrauterine death. PMID- 8725758 TI - Endometrial resection vs. abdominal hysterectomy for menorrhagia. Correlated sample analysis. AB - OBJECTIVE: To compare hysteroscopic endometrial resection with transabdominal hysterectomy in women with menorrhagia who failed to respond to conservative treatment. STUDY DESIGN: In order to attain correlated samples, only patients with a normal-sized or moderately enlarged uterus were included in the study. RESULTS: The operating time, hospital stay and number of women requiring postoperative blood transfusion was significantly lower in the hysteroscopy group. Following hysteroscopy, uterine perforation occurred in three patients, fluid overload in two and dilutional hyponatremia in one. Laparotomy was performed for suspected bowel injury in two cases. The main complication in women undergoing hysterectomy was postoperative fever (36.5%). One case of vesicovaginal fistula was diagnosed seven days postoperatively. CONCLUSION: The results indicate that endometrial resection, with its low cost and prompt recovery, offers a promising alternative treatment for women with abnormal uterine bleeding. PMID- 8725759 TI - Order of rotation in third-year clerkships. Influence on academic performance. AB - OBJECTIVE: To determine if student performance during the third-year clerkship is influenced by the order of departmental rotation. STUDY DESIGN: In this retrospective, descriptive study, the National Board of Medical Examiners (NBME) scores, departmental examination grade (DE) and final block grade (FG) were stratified into six segments based on the order of block time in obstetrics gynecology, pediatrics, psychiatry, internal medicine, surgery and family medicine. Consistency was validated by comparing these performance criteria from 1988 to 1992. RESULTS: When all six clerkships were assessed by year, there was no uniform trend of improved performance over time. In the obstetrics-gynecology block, however, NBME scores and FGs were highest for students assigned to the fifth and sixth rotation as compared to the first two groups (P < .02). No such trend was noted with the departmental written examination. In surgery, DE and FG were always lower in the first rotation (P = .0001). Psychiatry DE scores were significantly higher on the second or third blocks as compared to the fifth and sixth rotation positions (P < .001). Students had a higher FG in pediatrics when the sixth position was compared to the second block (P = .02), but the DE grades and NBME scores were not altered by rotational schema. In family medicine and internal medicine, no effect of rotation was noted. CONCLUSION: Third-year clerks tended to have higher NBME scores and FGs in obstetrics-gynecology if they selected this rotation later in the academic year. Surgery DEs and FGs were lowest if selected first in the academic year. There was no similar trend in other nonsurgical specialties. These data suggest that prior clinical rotations positively influence student performance in obstetrics-gynecology. PMID- 8725760 TI - Pelvic inflammatory disease. Correlation of severity with CA-125 levels. AB - OBJECTIVE: To determine whether elevated levels of cancer antigen 125 (CA-125) correlate with the severity and clinical outcome of pelvic inflammatory disease. STUDY DESIGN: CA-125 levels were measured prospectively in 36 women with pelvic inflammatory disease using a commercial immunoassay. Initial inclusion criteria were abdominal tenderness, cervical motion tenderness and adnexal tenderness. Patients then were categorized into three groups based on the severity of the illness. Group 1 included patients with (1) temperature > or = 38.0 but < 38.5 degrees C or (2) WBC count > or = 10 but < 15 x 10(3) cells/microL. Group 2 included patients with (1) temperature > or = 38.5 degrees C or (2) WBC count > or = 15 x 10(3) cells/microL. Group 3 included patients who fulfilled the same criteria as group 2 but who also had an adnexal mass. Differences in CA-125 levels were compared statistically using the unpaired Student t test. RESULTS: All patients with the diagnosis of pelvic inflammatory disease had CA-125 levels > 30 U/mL, with a range of 48-656. Moreover, the levels were significantly different among the three groups (group 1 vs. 2, P = .002; 1 vs. 3, P = .001; 2 vs. 3, P = .001), and a correlation between severity of disease and CA-125 levels was found. All patients with a presumed pelvic abscess had CA-125 levels in the range for neoplastic processes. CONCLUSION: Elevated CA-125 levels correlate with the severity of pelvic inflammatory disease and outcome. Pelvic inflammatory disease must be considered in the differential diagnosis of elevated CA-125 levels and pelvic masses. PMID- 8725762 TI - Subinvolution of the placental bed after molar pregnancy. AB - OBJECTIVE: To describe the occurrence of subinvolution of the placental bed following molar pregnancy. STUDY DESIGN: A review of archival histology and clinical charts. RESULTS: Four cases of subinvolution of the placental bed after a molar pregnancy were found in an institutional review; three were preceded by a complete mole and one by a partial mole. In all four cases the arteries were widely distended and mainly devoid of thrombus formation. Plentiful persistent interstitial trophoblast was seen within the decidua and myometrium, and endovascular trophoblast was seen embedded within the arterial walls in all four and intraluminally in two of the cases. CONCLUSION: The condition may be underdiagnosed after molar pregnancy and is a likely cause of continued bleeding following evacuation of the hydatidiform mole. PMID- 8725761 TI - Comparison of magnetic resonance imaging and transvaginal ultrasonography with hysterosalpingography in the evaluation of women exposed to diethylstilbestrol. AB - OBJECTIVE: To compare the ability of magnetic resonance imaging (MRI) and transvaginal ultrasound (TV-US) with that of hysterosalpingography (HSG) in detecting uterine abnormalities caused by in utero diethylstilbestrol (DES) exposure. STUDY DESIGN: The study was a prospective MRI and TV-US blind comparison of DES-exposed and nonexposed subjects who had had previous HSG for infertility evaluation. RESULTS: MRI identified uterine constrictions in 60% of patients and T-shaped uteri in 25% of DES-exposed patients with HSG-confirmed abnormalities. TV-US did not identify uterine constrictions or T-shaped uteri in DES-exposed patients. CONCLUSION: HSG must still be considered the preferred method in evaluating DES-related uterine abnormalities. HSG-defined uterine abnormalities associated with in utero DES exposure were variably identified by MRI and not at all by TV-US. PMID- 8725763 TI - Sexual function after sacrospinous ligament fixation for vaginal vault prolapse. AB - OBJECTIVE: To determine if sacrospinous ligament fixation predisposes to dyspareunia or if vaginal narrowing secondary to repair of associated defects is causative. STUDY DESIGN: We interviewed and examined 36 heterosexual patients preoperatively and at follow-up 15-79 months postoperatively. We inquired specifically as to sexual activity or inactivity and whether sexual function had improved or declined since surgery. RESULTS: Postoperatively, sexually active patients reported either an improvement or no change in sexual function unless vaginal narrowing occurred. Vaginal narrowing caused apareunia in three postoperative patients. Death of the partner was the most common reason for sexual inactivity preoperatively or postoperatively. CONCLUSION: Sacrospinous ligament fixation did not predispose to dyspareunia unless vaginal narrowing due to repair of associated defects was present. PMID- 8725764 TI - Massive ovarian edema in a menopausal woman. A case report. AB - BACKGROUND: Massive ovarian edema is a tumorlike condition in which there is marked enlargement of one or both ovaries due to accumulation of edema fluid in the stroma. It is speculated to occur as a result of partial ovarian torsion. CASE: Massive ovarian edema occurred in a postmenopausal woman. CONCLUSION: Massive ovarian edema is a rare cause of an adnexal mass. Knowledge of its existence is important to prevent incorrect treatment. We present the first case occurring in a menopausal woman. PMID- 8725765 TI - Leuprolide acetate in advanced ovarian serous tumor of low malignant potential. A case report. AB - BACKGROUND: Patients with advanced ovarian tumors of low malignant potential have no defined systemic therapy. Leuprolide acetate and other gonadotropin-releasing hormone analogues have definitive activity in ovarian epithelial carcinomas. On the basis of these considerations, we used leuprolide acetate to treat a young woman with an advanced ovarian tumor of low malignant potential. CASE: In a case of a long-term survivor of high-risk advanced-stage ovarian serous tumor of low malignant potential, suboptimal primary cytoreductive surgery was performed; at that time it was found that the tumor had directly invaded the outer uterine wall and rectosigmoid colon serosa and had metastasized to the pelvic lymph nodes. Postoperative adjunctive hormonal treatment with leuprolide acetate was administered and resulted in a complete clinical and serologic response. The patient remained disease free 68 months after the initial diagnosis. Her only symptoms were minimal hot flashes and vaginal dryness. CONCLUSION: Leuprolide acetate may be active in the treatment of ovarian tumors of low malignant potential. It should be considered frontline systemic therapy for this disease. PMID- 8725766 TI - Identification of a subtle chromosomal translocation in a family with recurrent miscarriages and a child with multiple congenital anomalies. A case report. AB - BACKGROUND: Couples who have had multiple miscarriages are at risk for carrying a balanced translocation since these carriers may produce unbalanced gametes. Small imbalances may lead to offspring with multiple congenital anomalies. This report emphasizes the importance of obtaining cytogenetics studies in couples with recurrent spontaneous abortions. CASE: A couple was referred for cytogenetic prenatal testing because of a history of recurrent miscarriages and an infant who died at 6 weeks of age with multiple congenital anomalies. Although the parental chromosomes were previously reported to be normal in another laboratory, the pedigree was consistent with a chromosomal etiology, and parental blood samples were reevaluated. The father was found to carry a subtle reciprocal translocation t(7;11)(q35;q23.3). Slides were obtained from the previous miscarriages and the infant who died. On reexamination, one miscarriage and the infant were found to be chromosomally unbalanced, carrying the derivative 7, resulting in partial monosomy for 7q and partial trisomy for 11q. The other miscarriage had a chromosomally normal female karyotype. Maternal cell contamination could not be excluded in that case. The current pregnancy was found to carry the balanced translocation. Since the rearrangement was quite small and subtle, fluorescence in situ hybridization (FISH) using "painting" probes for chromosomes 7 and 11 was used to confirm the balanced state in the fetus. CONCLUSION: This family illustrates the importance of performing high-quality chromosome studies on people who have spontaneous abortions and children with multiple congenital anomalies. The use of FISH probes was helpful in confirming this subtle rearrangement. PMID- 8725767 TI - Unanticipated pregnancy with intrauterine growth retardation after radiation induced ovarian failure. A case report. AB - BACKGROUND: Ovarian failure is common after pelvic irradiation and is dependent upon radiation dose and patient age. This case report demonstrates the resumption of ovulation and pregnancy subsequent to this diagnosis. CASE: An enlarging abdominal mass was noted in a 28-year-old female 20 months after resection of a pelvic hemangiopericytoma. She had received postoperative adjuvant hemipelvic radiotherapy and subsequently developed amenorrhea and symptoms of hypoestrogenism. Serum follicle-stimulating hormone (FSH) was elevated. In light of the diagnosis of ovarian failure, the finding of an intrauterine pregnancy on an abdominal computed tomography scout film, performed to rule out a recurrence of the primary tumor, was unexpected. CONCLUSION: While amenorrhea and elevation in serum gonadotropin levels are common after pelvic irradiation, the clinician must be cognizant of the potential for resumption of ovulation after radiotherapy. The diagnosis of ovarian failure should be based on more than a single serum FSH level. Further, radiation changes in the endometrium and myometrium as well as in uterine blood flow may have an adverse effect on pregnancy outcome. We suspect these effects had an etiologic influence on the fetal growth retardation in this pregnancy. PMID- 8725768 TI - Pyomyoma after uterine instrumentation. A case report. AB - BACKGROUND: Since 1945, only 10 cases of pyomyoma have been described in the literature. Four were related to pregnancy but none to elective abortion or uterine instrumentation. CASE: A 32-year-old woman was admitted to the hospital with a history of lower abdominal pain, nausea, vomiting and low grade fever with a past medical history significant for uterine leiomyomata and a second-trimester elective abortion 10 weeks prior to admission. Pelvic sonography demonstrated an enlarged uterus with a mass consisting of cystic and solid components. Uterine curettage revealed blood clots and scanty endometrial tissue. After no improvement following antibiotic therapy and nondiagnostic uterine curettage, an exploratory laparotomy was performed. It revealed peritonitis with multiple pyomyomas draining purulent material. CONCLUSION: Although rarely reported, pyomyoma should be considered in the setting of a recent history of uterine instrumentation, signs of infection and a uterine myoma. PMID- 8725769 TI - Information retrieval and genomics--an introduction. PMID- 8725770 TI - A graph conceptual model for developing Human Genome Center databases. AB - We have developed a representation of genome data which has proven itself useful for describing data at a Human Genome Center. Genomic data have a graph-like structure and representing the concepts and relationships of genetics as a graph simplifies the development of databases for genome laboratories. Graphs are a comfortable communication medium for biologists and computer scientists and graph diagrams assist in the development of databases by facilitating the exchange of expertise. We have tailored a graph language for modeling genomic data and describe our process of using graphs to develop genome databases. PMID- 8725771 TI - PROXIMAL: a database system for the efficient retrieval of genetic information. AB - We report on a query language, called PROXIMAL, for the efficient specification of complex queries over biological sequences. We present a data model for asking complex and structured questions of biological sequences, and show how the query language can be used. In addition, we describe a visual query metaphor intended for a biologist-friendly front end to the PROXIMAL query language. PMID- 8725772 TI - An analysis of statistical term strength and its use in the indexing and retrieval of molecular biology texts. AB - The biological literature presents a difficult challenge to information processing in its complexity, diversity, and in its sheer volume. Much of the diversity resides in its technical terminology, which has also become voluminous. In an effort to deal more effectively with this large vocabulary and improve information processing, a method of focus has been developed which allows one to classify terms based on a measure of their importance in describing the content of the documents in which they occur. The measurement is called the strength of a term and is a measure of how strongly the term's occurrences correlate with the subjects of documents in the database. If term occurrences are random then there will be no correlation and the strength will be zero, but if for any subject, the term is either always present or never present its strength will be one. We give here a new, information theoretical interpretation of term strength, review some of its uses in focusing the processing of documents for information retrieval and describe new results obtained in document categorization. PMID- 8725773 TI - Ethics, genomics, and information retrieval. AB - The union of genomics and computational information retrieval raises a number of ethical issues, including data sharing, database accuracy, group and subgroup stigma, and privacy and confidentiality. These issues are introduced and assigned a preliminary analysis which, it is hoped, may be of use in more sustained efforts to identify issues, solutions and potential guidelines, to stimulate education, and to strike the most appropriate balance between the rights of individuals and the needs of researchers and society. PMID- 8725775 TI - The effects of intra-aortic balloon pumping combined with coronary sinus interventions on failing hearts: a model study. AB - Cardiac energy balance is an important factor in evaluating cardiac function. In this work, the effects of different assist modalities and their combinations on cardiac energy balance and global cardiac function have been investigated. The simulation results indicate that coronary sinus interventions (CSIs) can provide myocardium with more energy than IABP when coronary arterial stenosis is serious. However, IABP can decrease myocardial energy consumption and enhance cardiac output, which cannot be done by CSIs, under the condition of impaired myocardial contractility. Therefore, we conclude that the combination of IABP and CSIs is the best way of assisting failing circulation. PMID- 8725774 TI - Engineering a normally responsive bioartificial pancreas based on glucose hypersensitive cells. AB - An implantable bioartificial pancreas is promising for diabetes treatment; transformed cells may allow the fabrication of these tissue constructs at a medically relevant scale. However, transformed cells are generally glucose hypersensitive, i.e. insulin secretion is near maximal at much lower than physiological glucose levels. The development of a normally responsive construct based on hypersensitive cells was examined. Two systems of immobilized cells and glucose-consuming enzyme were considered: cells surrounded by the enzyme, and cells coimmobilized with the enzyme. Model simulations showed that, with properly chosen parameter values, the first system can mimic the response of normal islets. PMID- 8725776 TI - Left ventricular ejection: model solution by collocation, an approximate analytical method. AB - A differential equation solution method that does not utilize numerical integration is demonstrated on a physiologic model. A previously proposed model of left ventricular ejection incorporating time-varying elastance, internal resistance, aortic inductance, and a three-component windkessel is solved by collocation. Assuming internal resistance to be constant allows simplification to a third order linear differential equation in left ventricular volume. A trigonometric series is used to approximate the solution and the coefficients of the series as well as the duration of ejection and pre-ejection periods are chosen so that the governing differential equation is exactly satisfied at certain times during ejection (the collocation points), as well as the boundary conditions and steady state condition. PMID- 8725777 TI - Quickbasic program for exact and mid-P confidence intervals for a binomial proportion. AB - Computation of the exact confidence interval for a binomial proportion is tedious and requires iterative computer techniques. A Quickbasic program for the IBM PC computer is described that accomplishes the confidence interval calculation accurately, quickly, and with no limits placed on the confidence level, or the size of the numerator or denominator. If desired, the mid-P confidence interval can be computed. The compiled program takes one second on the IBM PC 386/33 without a math co-processor. PMID- 8725778 TI - Image analysis of tissue sections. AB - The use of computers for the automated image analysis of tissue sections is becoming increasingly important. The paper presents an overview of current methodologies and summarizes developments in this field. A brief introduction followed by a survey is provided in each of these areas: image transformation, image segmentation and classification. PMID- 8725779 TI - Genetic drift. Prying open the box. PMID- 8725780 TI - Diffuse cystic renal dysplasia: nonsyndromal familial case. AB - We report on a family in which three individuals, a male and two females were affected with nonsyndromal diffuse cystic dysplasia of the kidneys. The parents had no renal abnormality. The occurrence of diffuse cystic dysplasia in three sibs born to normal parents suggests autosomal recessive inheritance. PMID- 8725781 TI - Leiomyoma of uterus in a patient with ring chromosome 12: case presentation and literature review. AB - We report on a 30-year-old women with de novo ring chromosome 12 mosaicism, 46,XX, r(12)(p13.3q24.3)/46,XX. In addition to the clinical manifestations generally observed in "ring syndrome" cases such as growth retardation, short stature, microcephaly, and mental deficiency, she had a broad nasal bridge, micrognathia with overbite, underdeveloped breasts, mild dorsal scoliosis, clinodactyly of the fifth fingers with a single interdigital crease, symphalangism of thumbs, tapering fingers, mild cutaneous syndactyly between the second and third toes, multiple cafe-au-lait spots, sebaceous acne on the face and back, and mild dystrophic toenails. She developed a large, pedunculated uterine leiomyoma at age 28 years. To our knowledge, uterine leiomyoma in association with r(12) has not been reported previously. However, a gain of chromosome 12 and translocations involving 12q14-15 have been described. PMID- 8725782 TI - Comparison of the canine and human acid beta-galactosidase gene. AB - Several canine cDNA libraries were screened with human beta-galactosidase cDNA as probe. Seven positive clones were isolated and sequenced yielding a partial (2060 bp) canine beta-galactosidase cDNA with 86% identity to the human beta galactosidase cDNA. Preliminary analysis of a canine genomic library indicated conservation of exon number and size. Analysis by Northern blotting disclosed a single mRNA of 2.4 kb in fibroblasts and liver from normal dogs and dogs affected with GM1 gangliosidosis. Although incomplete, these results indicate canine GM1 gangliosidosis is a suitable animal model of the human disease and should further efforts to devise a gene therapy strategy for its treatment. PMID- 8725783 TI - Severe facial clefting, limbic dermoid, hypoplasia of the corpus callosum, and multiple skin appendages: severe frontofacionasal "dysplasia" or newly recognised syndrome? AB - We report on a child with severe midline facial cleft, bilateral cleft lip and palate, telecanthus, S-shaped palpebral fissures, limbic dermoid, midface hypoplasia, hypoplastic corpus callosum, and multiple skin appendages. This case may be an example of severe frontofacionasal "dysplasia" or a newly recognised syndrome. PMID- 8725784 TI - Molecular analysis in true hermaphrodites with different karyotypes and similar phenotypes. AB - True hermaphroditism is characterized by the development of ovarian and testicular tissue in the same individual. Mullerian and Wolffian structures are usually present, and external genitalia are often ambiguous. The most frequent karyotype in these patients is 46,XX or various forms of mosaicism, whereas 46,XX is very rarely found. The phenotype in all these subjects is similar. We studied 10 true hermaphrodites. Six of them had a 46,XX chromosomal complement: 3 had been reared as males and 3 as females. The other 4 patients were mosaics: 3 were 46,XX/46,XY and one had a 46,XX/47,XXY karyotype. One of the 46,XX/46,XY mosaics was reared as a female, whereas the other 3 mosaics were reared as males. The sex of assignment in the 10 patients depended only on labio-scrotal differentiation. Molecular studies in 46,XX subjects documented the absence of Y centromeric sequences in all cases, arguing against hidden mosaicism. One patient presented Yp sequences (ZFY+, SRY+), which contrast with South African black 46,XX true hermaphrodites in whom no Y sequences were found. Molecular analysis in the subjects with mosaicism demonstrated the presence of Y centromeric and Yp sequences confirming the presence of a Y chromosome. Gonadal development, endocrine function, and phenotype in the 10 patients did not correlate with the presence of a Y chromosome or Y-derived sequences in the genome, confirming that true hermaphroditism is a heterogeneous condition. PMID- 8725785 TI - Urinary bile acids and peroxisomal bifunctional enzyme deficiency. AB - The biosynthesis of normal bile acids involves beta-oxidation of the 8-carbon side-chain of cholesterol, in addition to numerous modifications of the sterol nucleus. Because beta-oxidation of the sterol side-chain has been localized to the peroxisome, bile acid analysis has been suggested to be useful in the diagnostic evaluation of individuals suspected of having peroxisomal disorders. Although data from subjects with generalized peroxisomal disorders support this, few data exist regarding the bile acids in individuals having single peroxisomal beta-oxidation enzyme disorders. In this study, we analyzed the urinary bile acids from 12 patients with peroxisomal bifunctional protein deficiency using continuous flow fast atom bombardment mass spectrometry. All 12 patients had abnormal spectra, although their ion profiles and rank order of intensity of ions varied considerably. Ten of 12 individuals had abnormal spectra with presence of taurine-conjugated tetrahydroxycholestenoates, allowing a definite diagnosis of a peroxisomal beta-oxidation defect and a presumptive diagnosis of bifunctional protein deficiency; the other two cases were nondiagnostically abnormal. The strengths and limitations of urinary bile acid analysis for the diagnosis of peroxisomal beta-oxidation disorders are discussed. PMID- 8725786 TI - Follow-up of a familial translocation t(10;16) with an unusual segregation pattern. AB - Bofinger et al. [Am J Med Genet 38:1-8, 1991] reported on a four-generation family with an unusual segregation pattern involving a translocation t(10;16)(q26.3;p13.1). All relatives either had a balanced or unbalanced translocation. We report on five additional relatives, none of whom have a normal karyotype. This unusual segregation pattern may be due to chance or be the result of meiotic drive. PMID- 8725787 TI - Molecular and cytogenetic characterization of a recurrent unbalanced translocation (4;21)(p16.3;q22.1): relevance to the Wolf-Hirschhorn and Down syndrome critical regions. AB - We report on an aneuploidy syndrome due to the unbalanced segregation of a familial translocation (4;21)(p16.3;q22.1) causing a partial 4p monosomy and a partial 21q trisomy. The three affected children presented with severe failure to thrive, short stature, microcephaly, profound hypotonia, and mental retardation. The face, very similar in the three children, is characterized by frontal bossing, upslanting of the palpebral fissures, short nose, and deep set ears, giving the overall appearance of the Down syndrome. The molecular study has defined the aneuploid segment on both 4p and 21q. Most of the Down syndrome critical region was found to the trisomic, while only part of the candidate Wolf Hirschhorn syndrome critical region was deleted, suggesting that this region is not critical for the major malformations characteristic for WHS. PMID- 8725788 TI - Two patients with duplication of 17p11.2: the reciprocal of the Smith-Magenis syndrome deletion? AB - J.M. and H.G. are two unrelated male patients with developmental delay. Cytogenetic analysis detected a duplication of 17p11.2 in both patients. The extent of the duplicated region was determined using single copy DNA probes: cen D17S58-D17S29-D17S258-D17S71-D17S445-+ ++D17S122-tel. Four of the six markers, D17S29, D17S258, D17S71, and D17S445, were duplicated by dosage analysis. Fluorescent in situ hybridization (FISH) analysis of H.G., using cosmids for locus D17S29, confirmed the duplication in 17p11.2. Because the deletion that causes the Smith-Magnesis syndrome involves the same region of 17p11.2 as the duplication in these patients, the mechanism may be similar to that proposed for the reciprocal deletion/duplication event observed in Hereditary Neuropathy with Liability to Pressure Palsies (HNPP) and Charcot-Marie-Tooth Type 1A disease (CMT1A). PMID- 8725789 TI - Novel findings in a patient with Weaver or a Weaver-like syndrome. AB - We report on a young male patient with an overgrowth syndrome, who had normal birth weight. He had a number of manifestations typical of the Weaver syndrome (WS), such as advanced bone age, peculiar craniofacial appearance, and camptodactyly. He also showed severe mental and speech retardation and demineralisation of the bones of the hands and feet. The latter can be considered as unreported manifestations of WS, or the patient could represent an example of a new WS-like syndrome. PMID- 8725790 TI - Cardio-facio-cutaneous (CFC) syndrome: report of an adult without mental retardation. AB - We report on a 25-year-old woman with typical manifestations of the cardio-facio cutaneous (CFC) syndrome, but without mental retardation. She had valvular and infundibular pulmonic stenosis, brittle and woolly hair with patchy alopecia, scant body hair, dry and hypohydrotic skin, and characteristic facial traits. To our knowledge, this is the first case of CFC syndrome without mental retardation but typical cutaneous findings. PMID- 8725791 TI - Estimating parental relationship in linkage analysis of recessive traits. AB - In linkage analysis of recessive traits, parental relationship is important. For the case that it is unknown, the question is investigated as to whether estimating parental relationship and using the estimated relationship in linkage analysis is beneficial. Results show that estimating parental relationship can reliably be carried out on the basis of 50-100 genetic marker loci (analysis based on theory by Thompson [1975: Am J Hum Genet 39:173-188]). Misspecification of parental relationship leads to a loss of linkage informativeness, but not to false-positive evidence for linkage. An asymptotic bias in the recombination fraction estimate occurs when parents are unrelated and falsely taken to be related, but no such bias is seen when related parents are taken to be unrelated. Results from this investigation suggest that an estimated parental relationship may be used in linkage analysis as if it were the correct relationship, when evidence for the estimated relationship is supported by a likelihood ratio of at least 10:1 against the parents being unrelated. PMID- 8725792 TI - A complex five breakpoint intrachromosomal rearrangement ascertained through two recombinant offspring. AB - Intrachromosomal rearrangements usually result from three of fewer breaks. We report a complex intrachromosomal rearrangement resulting from five breaks in one chromosome 10 of a phenotypically normal father of two developmentally delayed children. GTG-banding analysis of the father's rearranged chromosome 10 suggested in initial pericentric inversion followed by an insertion from the short arm into the terminal band of the long arm [der(10) (pter-->p13::q21.2-->p12.2::q22.1::- >q26.3::q22.1-->q 21.2::p12.2-->p13::q26.3-->qter)]. To our knowledge, this rearrangement is the most complex ever reported in a single chromosome. Both children inherited a recombinant chromosome 10 with loss of the insertion and the segment distal to it [rec(10)der(pter-->p13: :q21.2-->p12.2::q22.1-->q26.3:)]. Mechanisms for both rearrangements are proposed. PMID- 8725793 TI - Fragile X syndrome in two siblings with major congenital malformations. AB - We report on 2 brothers with both fragile X and VACTERL-H syndrome. The first sibling, age 5, had bilateral cleft lip and palate, ventricular septal defect, and a hypoplastic thumb. The second sibling, age 2 1/2, had a trachesophageal fistula, esophageal atresia, and vertebral abnormality. High-resolution chromosome analysis showed a 46, XY chromosome constitution in both siblings. By PCR and Southern blot analysis, the siblings were found to have large triplet repeat expansions in the fragile X gene (FMR 1) and both had methylation mosaicism. Enzyme kinetic studies of iduronate sulfatase demonstrated a two-fold increase in activity in the first sib as compared to the second. Possible mechanisms through which the fragile X mutation can cause down-regulation of adjacent loci are discussed. PMID- 8725794 TI - In vitro studies on clonal growth of chondrocytes in thanatophoric dysplasia. AB - Thanatophoric dysplasia (TD) is characterized by a disorganized growth plate with markedly reduced proliferative and hypertrophic cartilage zones. Therefore, we studied in vitro the proliferation rates of articular chondrocytes from five TD patients and age-matched controls in response to bFGF, IGF-I, IGF-II, and TGF beta 1. In human fetal controls bFGF was the most potent growth factor. Clonal growth the articular chondrocytes in response to bFGF was reduced in two of five TD patients and slightly below the range of controls in a third case. Stimulation of chondrocyte proliferation by IGF I and II was reduced in the patient whose response to bFGF was most markedly impaired. The effect of TGF-beta 1 ranged from normal to slightly elevated values in TD fetuses. These results indicate heterogeneity of the underlying defects in TD. Low proliferative responses of chondrocytes to bFGF and IGF-I/II are likely to play a key role in the pathogenesis of some cases. In two of five patients studied, the mechanisms of bFGF and IGF-signal transduction are candidates for the primary molecular defect. PMID- 8725795 TI - Natural history study of pseudoachondroplasia. AB - Pseudoachondroplasia (PSACH) is a well-characterized autosomal dominant dwarfing condition. A great deal of information is available about orthopedic complications, but little is known about extraskeletal complications in adulthood. This study was undertaken to delineate the natural history of PSACH at all ages. Seventy-nine individuals responded to an extensive questionnaire that included information about deformities, operations, general health, chronic diseases, and reproduction. PSACH individuals were ascertained through the University of Texas Medical Genetics patient population, a genetic linkage study, and the social organization, Little People of America. The results show that PSACH individuals with a family history do not have a distinct or more severe phenotype than new mutation cases. There were not differences in the number of orthopedic complications, operations, or number of offspring between these two groups. Less than half of affected adults reported having total hip replacement surgery, which was less common than previously reported. Extraskeletal complications were generally uncommon. There were four cases of cancers in 41 individuals queried. Premature osteoarthritis was the major health problem for PSACH individuals. PSACH individuals are generally healthy but have problems associated with debilitating osteoarthritis. PMID- 8725796 TI - Diaphragmatic and multiple midline defects. PMID- 8725797 TI - Familial Dandy-Walker variant in CDG syndrome. PMID- 8725798 TI - Detection of imprinting mutations in Angelman syndrome using a probe for exon alpha of SNRPN. PMID- 8725799 TI - Comparison of polymerase chain reaction and microbiological culture for detection of salmonellae in equine feces and environmental samples. AB - OBJECTIVE: To compare the sensitivity of polymerase chain reaction (PCR) with microbiological culture for detecting salmonellae in equine fecal samples and equine environmental swab specimens. DESIGN: Samples and specimens were tested by PCR and microbiological culture. SAMPLE POPULATION: A fecal sample from each of 152 horses admitted consecutively to the clinic for evaluation by the outpatient service, 282 fecal samples from 110 hospitalized horses that had been submitted to the clinical microbiology laboratory, and 313 environmental swab specimens were examined. PROCEDURE: Each sample and specimen in the study was tested, using PCR and microbiological culture. Results of PCR and culture were compared. RESULTS: Significantly (P < 0.001) more fecal samples were positive by PCR than by microbiological culture. 26 of 152 (17.1%) fecal samples collected from horses admitted by the outpatient service were positive by PCR and none was positive by culture. 71 of 110 hospitalized horses were identified as positive by PCR, compared with 11 horses identified as positive by culture. All culture-positive horses were positive by PCR. Of the 11 culture-positive horses, 10 (90.9%) were identified as PCR positive after testing of the first sample submitted, compared with 7 (63.6%) by culture. All PCR-positive horses were detected after a total of 3 samples/horse were submitted, whereas as many as 5 samples/horse was required to identify all culture-positive horses. 8 of 313 environmental specimens were positive by PCR, and none was positive by culture. CONCLUSION: The PCR method reported here was more sensitive, more rapid, and required submission of fewer samples or specimens than did microbiological culture for detecting salmonellae. PMID- 8725800 TI - Effects of various methods of preservation on the stability of uric acid in frozen canine urine. AB - OBJECTIVE: To evaluate the effects of dilution and alkalinization, separately and together, on the stability of uric acid in canine urine stored at -20 C. DESIGN: Prospective-controlled study. ANIMALS: 5 dogs with confirmed ammonium urate uroliths, 6 Beagles, and 6 mixed-breed dogs. PROCEDURE: Dogs were fed a 31.4% protein (dry weight), meat-based diet for 21 days, and urine samples were collected on day 22. Urine samples were preserved, using combinations of dilution and alkalinization, and divided into 1-ml aliquots for storage at -20 C for 1 to 12 weeks. Urine uric acid concentrations were measured, using high-performance liquid chromatography, on day of collection (baseline), and after 1, 2, 4, 8, and 12 weeks. RESULTS: Alkalinization did not have a significant effect on reproducibility of measurements of uric acid concentrations in urine; however, dilution did have a significant effect. Compared with baseline, uric acid concentrations in urine samples collected from dogs with ammonium urate uroliths and Beagles and diluted 1:10 or 1:20 with deionized water were not different after storage for 1 to 12 weeks. Uric acid concentrations in urine samples collected from mixed-breed dogs did not differ from baseline values during the 12 week storage period whether samples were undiluted or were diluted 1:10 or 1:20 with deionized water. CONCLUSIONS: Measurements of uric acid concentration are most reproducible in canine urine samples stored at -20 C for 1 to 12 weeks when samples are diluted 1:20 with deionized water. CLINICAL RELEVANCE: To ensure reproducibility of measurements of uric acid concentration in urine samples collected from dogs affected with urate uroliths, urine should be diluted 1:20 with deionized water. Alkalinization is not necessary, and is not recommended because of the additional step in processing and its potential to interfere with measurement of other urinary analytes. PMID- 8725801 TI - Mitral valve protrusion assessed by use of B-mode echocardiography in dogs with mitral regurgitation. AB - OBJECTIVE: To examine our hypothesis that changes of the mitral valve in dogs with mitral regurgitation lead to various degrees of protrusion of the cusp in individual dogs in systole. DESIGN: Assessing the mitral protrusion, using B-mode echocardiography in dogs with mitral regurgitation. ANIMALS: 33 dogs with chronic mitral regurgitation and 40 clinically normal dogs. PROCEDURE: The mitral valve was imaged in the right parasternal left ventricular long-axis view, and we measured the height (H) from the coaptation point or tip of the protruded cusp to the mitral annular plane. When the tip of the protruded cusp was located beyond the mitral annular plane extending into the left atrium, H was expressed as negative values. RESULTS: The H in clinically normal dogs was 3.7 +/- 0.8 mm. In dogs with mitral regurgitation, the mitral cusp protruded toward the left atrium to various degrees, and H was -0.7 +/- 1.9 mm (range, 2.9 to -5.0 mm). The 2 dogs in the latter group with the lowest H (-5.0 and -4.5 mm, respectively) presented typical mitral flail on B-mode images. The indices of left atrial and ventricular dimension (left atrial dimension/aortic dimension and left ventricular end diastolic dimension/body weight, respectively) were significantly higher than those in the clinically normal dogs and were negatively correlated with H. CONCLUSION: The mitral valve in individual dogs with mitral regurgitation has different degrees of protrusion. The degree of protrusion might be related to the hemo-dynamic condition in mitral regurgitation. PMID- 8725802 TI - Characterization of anatomic communications between the femoropatellar joint and lateral and medial femorotibial joints in cattle, using intra-articular latex, positive contrast arthrography, and fluoroscopy. AB - OBJECTIVE: To evaluate the frequency of communication between the lateral and medial femorotibial joints and the femoropatellar joint in cattle. DESIGN: 1 of 3 injection sites was randomly assigned to each ox. ANIMALS: 102 limbs were obtained from 55 fresh bovine cadavers presented for necropsy with problems unrelated to the stifle. PROCEDURE: The joints were injected with a mixture of latex and barium sulfate. Communication between 2 or more joints was determined by the presence of latex and contrast material in a joint adjacent to the injected joint by examining frozen sections and positive-contrast arthrography. RESULTS: Communication between the 3 joints was present in 58 (56.9%) limbs. The femoropatellar and the medial femorotibial joints always communicated. Thirteen of 38 (34.2%) specimens injected in the lateral femorotibial joint did not communicate with the 2 other joints. The femoropatellar joint communicated with the lateral and medial femorotibial joints on the distal abaxial aspect of the trochlear ridge. CONCLUSION: Individual anatomic variation of the stifle in cattle should be considered when diagnostic or treatment protocols are established. CLINICAL RELEVANCE: The lateral femorotibial joint should be treated separately because it does not consistently communicate with the femoropatellar or medial femorotibial joint. PMID- 8725803 TI - Calculation of maximal removal rate of indocyanine green to measure hepatic functional mass in dogs by use of the nine-hour method. AB - OBJECTIVE: To establish a versatile and reliable procedure for the determination of indocyanine green maximal removal rate (ICG Rmax) to measure hepatic functional mass in dogs within 9 hours (9-hour method). DESIGN: Relation between 9-hour and standard 3-day methods was examined. ANIMALS: 101 healthy dogs. PROCEDURE: On investigation of the optimal technical conditions allowing completion of all procedures in a day, the appropriate IV administered doses of ICG were 0.125, 0.5, and 2.0 mg/kg of body weight, and the best blood sample collection times for obtaining plasma half-life at these 3 doses were before and 3, 6, and 9 minutes after ICG administration. RESULTS: Comparison of the 9-hour with 3-day method yielded a correlation coefficient (r) of 0.84, indicating close (P < 0.01) correlation. In the 9-hour method, mean +/- SD of ICG Rmax in healthy dogs was 0.24 +/- 0.09 mg/kg/min in male (n = 62) and was 0.23 +/- 0.06 mg/kg/min in female (n = 21) Beagles, and was 0.21 +/- 0.10 mg/kg/min in male (n = 11) and 0.20 +/- 0.07 mg/kg/min in female (n = 7) mixed-breed dogs. In Beagles treated orally with carbon tetrachloride (0.1 ml/kg in gelatine capsules) thrice weekly during a 10-week period, plasma alanine transaminase activity plateaued at a high value (> 2,000 IU/L) on day 5, and remained at this value until the end of the study. The ICG Rmax changed accordingly: day 5, 0.17; day 10, 0.11; day 40, 0.05; and day 60, 0.06 mg/kg/min. CONCLUSION: The 9-hour method for determination of ICG Rmax correlates favorably with the 3-day method. CLINICAL RELEVANCE: This procedure may be practically applied in veterinary clinics in terms of prediction of hepatic functional mass, and for diagnosis of hepatotoxicosis induced by certain compounds. PMID- 8725805 TI - Viral status and antibody response in cattle inoculated with recombinant bovine leukemia virus-vaccinia virus vaccines after challenge exposure with bovine leukemia virus-infected lymphocytes. AB - OBJECTIVE: To determine the bovine leukemia virus (BLV) antibody response and infectivity status in BLV-vaccinated cattle after challenge exposure with BLV infected lymphocytes. DESIGN: Steers were inoculated with vaccinia virus constructs expressing the gag, pol, and env genes of the BLV or the env gene only of the BLV genome, then challenge exposed with BLV-infected lymphocytes. The steers' BLV antibody and infectivity status was monitored. ANIMALS: Fifteen 8- to 9-month-old Holstein steers previously determined to be BLV antibody and BLV negative. PROCEDURE: 1 month after second after inoculation, steers were challenge exposed with 10(6) BLV-infected lymphocytes from a highly infective BLV positive cow. Serum and blood lymphocytes were obtained regularly for 6 months. The agar gel immunodiffusion assay, ELISA, and serum neutralization assay were used to detect BLV antibody in serum of steers. The sheep infectivity and syncytium-forming assays were used to determine the viral status of the steers. RESULTS: Differences were seen in antibody responses between the BLV-vaccinated and non-BLV-vaccinated control groups. All cattle were susceptible to infection when challenge exposed with BLV-infected lymphocytes. CONCLUSION: Despite the enhanced immune response in the BLV-vaccinated cattle after challenge exposure, none of the BLV-vaccinated cattle was protected from BLV infection. CLINICAL RELEVANCE: Vaccination is not an effective way to protect cattle from BLV infection. PMID- 8725806 TI - Specific immune responses of dairy cattle after primary inoculation with recombinant bovine interferon-gamma as an adjuvant when vaccinating against mastitis. AB - OBJECTIVE: To examine whether specific immunity in the mammary gland and blood of dairy cattle was enhanced after primary inoculation with a test antigen keyhole limpet hemocyanin (KLH) and recombinant bovine interferon-gamma (rBoIFN-gamma) as an adjuvant. DESIGN: Holstein dairy cows received IM injections of KLH in conjunction with saline solution (n = 4), Freund's incomplete adjuvant (FIA; n = 4), or rBoIFN-gamma (n = 3). Milk and blood samples were collected during a 1 month period and analyzed for KLH antibody content. Isolated blood mononuclear cells were examined for their ability to proliferate and produce interleukin 2 (IL-2) after mitogen and/or KLH stimulation in vitro. The phenotype of isolated blood mononuclear cells also was characterized through flow cytometric analysis. RESULTS: The adjuvant rBoIFN-gamma induced serum KLH antibody titers similar to those induced by administration of FIA. However, FIA induced significantly more KLH antibodies in milk. Administration of rBoIFN-gamma enhanced T-lymphocyte activity, as indicated by the greater expression of high-affinity IL-2 receptors and the increased response to the mitogens concanavalin A, phytohemagglutinin, and IL-2, compared with FIA or saline treatment. Lymphocyte and monocyte movement from the blood also was altered after rBoIFN-gamma treatment, which can have a profound influence on secondary immune responses, such as antibody production. CONCLUSIONS: rBoIFN-gamma may safely enhance specific immunity in the bovine mammary gland and may be an effective adjuvant in mastitis immunization protocols. PMID- 8725804 TI - Prevalence of Serpulina species in relation to diarrhea and feed medication in pig-rearing herds in Sweden. AB - OBJECTIVE: To determine prevalence of various pheno- and genotypes of Serpulina sp in young pigs in relation to diarrhea and feed medication in Swedish pig rearing herds. DESIGN: Isolation of spirochetes. Phenotypical and genotypical classification. SAMPLE POPULATION: Young pigs (n = 358) in 19 pigrearing herds. PROCEDURE: Serpulina isolates were classified according to a biochemical scheme based on hemolysis, indole production, hippurate hydrolysis, and alpha galactosidase, alpha-glucosidase, and beta-glucosidase activities. The 16S rRNA sequences for 10 of the field strains and 2 type strains of Serpulina spp were aligned and compared. Minimum inhibitory concentrations of olaquindox for 9 of the strains were determined. RESULTS: Weakly beta-hemolytic intestinal spirochetes (WBHIS) were isolated from 17 of the herds and 65% of the samples. More than 1 phenotype of WBHIS was found in 12 of the 19 herds. S hyodysenteriae was not isolated in any of the herds. Hippurate-positive WBHIS were isolated in 6 of 7 herds affected by diarrhea, but in only 1 of 8 herds without diarrhea. Hippurate-positive strains were closely related to the pathogenic strain P43 if judged from sequence comparisons. Strains with the same biochemical profile isolated within a herd had identical sequences, but when isolated from different herds, sequence differences were observed. The prevalence of WBHIS was reduced in herds medicated with olaquindox. Investigated field strains had minimum inhibitory concentration values < or = 1 microgram/ml for olaquindox. CONCLUSION: The presence of WBHIS, with the ability to hydrolyze hippurate, was related to diarrhea in pig herds. CLINICAL RELEVANCE: Potentially pathogenic WBHIS can be distinguished from nonpathogenic strains by the hippurate hydrolysis test. PMID- 8725807 TI - Influence of vitamin D3 infusion and dietary calcium on secretion of interleukin 1, interleukin 6, and tumor necrosis factor in mice infected with Mycobacterium paratuberculosis. AB - OBJECTIVE: To study the effects of 1,25(OH)2D3 and calcium (Ca) on splenocyte cytokine secretion during Mycobacterium paratuberculosis infection. DESIGN: Mice were assigned to the following treatments: 1-noninfected, 2-infected, 3 noninfected/1,25(OH)2D3, 4-infected/1,25(OH)2D3, and 5-infected/low-Ca diet (0.15%). ANIMALS--Male beige mice averaging 6 weeks of age and 20 g in body weight. PROCEDURE: After acclimation to their diets, mice in treatments 2, 4, and 5 were inoculated IV with 10(8) colony-forming units of M paratuberculosis. At 1, 6, and 12 months after infection, mice in treatment groups 3 and 4 had miniosmotic pumps implanted subcutaneously that delivered 13 ng of 1,25(OH)2D3/day for 14 days. Treatment 5 was included as a control for comparison with treatment 4 because low dietary Ca should increase endogenous 1,25(OH)2D3 values. Splenocytes were isolated from mice at 1, 6, and 12 months and stimulated in vitro with medium alone (nonstimulated), lipopolysaccharide (LPS), concanavalin A, and M paratuberculosis whole-cell sonicate (MpS). RESULTS: Production of interleukin 6 after stimulation with LPS, concanavalin A, or MpS was higher (P < 0.05) for splenocytes isolated from mice fed the low-Ca diet, compared with control infected mice 1 and 6 months after infection. Interleukin 1 and tumor necrosis factor activities were increased (P < 0.05) in splenocytes cultured with LPS and MpS after isolation from mice of the low-Ca group. Mice infused with 1,25(OH)2D3 had higher (P < 0.05) interleukin 1 secretion after stimulation of splenocytes with LPS and higher (P < 0.05) tumor necrosis factor production after incubation with MpS. CONCLUSION: 1,25(OH)2D3 and low dietary Ca increase cytokine secretion in mice infected with M paratuberculosis. PMID- 8725808 TI - Morphologic changes and immunohistochemical detection of viral antigen in ovaries from cattle persistently infected with bovine viral diarrhea virus. AB - OBJECTIVE: To identify morphologic differences in ovaries from cows persistently infected with bovine viral diarrhea virus (BVDV) and determine ovarian cell types infected in these cows. DESIGN: A comparative study of ovaries in cattle persistently infected with BVDV and cattle not persistently infected with BVDV, using morphologic and immunohistochemical analysis. ANIMALS: 6 postpubertal cows persistently infected with BVDV and 6 postpubertal cows not persistently infected with BVDV. PROCEDURE: Ovaries were compared morphologically by counting the number of normal structures present on 3 histologic sections taken from each ovary. Immunohistochemistry was accomplished, using an indirect, monoclonal antibody-linked, avidin-biotin-peroxidase complex procedure. RESULTS: Significant (P < 0.01) decrease in the number of tertiary follicles, graafian follicles, atretic follicles, and corpus hemorrhagicum/luteum/albicans was observed in cows persistently infected with BVDV. No difference in numbers of primordial or secondary follicles was observed. Immunostaining of BVDV antigen in luteal cells and macrophage-like cells was evident in ovaries from cows persistently infected with BVDV. CONCLUSIONS: Cows persistently infected with BVDV appear to have significant morphologic changes in their ovaries that suggest reduction in normal ovarian activities. Furthermore, BVDV antigen can be identified in specific ovarian cell types in cattle persistently infected with BVDV. CLINICAL RELEVANCE: The changes observed may reduce reproductive performance in cows persistently infected with BVDV, and may be of importance when trying to salvage valuable genetic material from persistently infected cows through embryo transfer. It is yet to be determined whether similar findings are true in cows acutely infected with BVDV. PMID- 8725809 TI - Comparison among strains of porcine reproductive and respiratory syndrome virus for their ability to cause reproductive failure. AB - OBJECTIVE: To compare the virulence of selected strains of porcine reproductive and respiratory syndrome virus (PRRSV) relative to reproductive performance of pregnant gilts. DESIGN: 16 pregnant gilts (principals) were exposed oronasally to 4 strains (vaccine strain RespPRRS, field strains VR-2385, VR-2431, and NADC-8, 4 gilts/strain) of PRRSV on or about day 90 of gestation, 4 pregnant gilts (controls) were kept under similar conditions, except for exposure to PRRSV. Samples and specimens obtained from gilts, pigs (before ingestion of colostrum), and fetuses were tested for PRRSV and homologous antibody. ANIMALS: 20 pregnant gilts. PROCEDURE: The virulence of each strain of PRRSV was evaluated mainly on the clinical status of the corresponding litters at farrowing. RESULTS: Most gilts remained clinically normal throughout the study and farrowed normally at or near the expected farrowing time. All virus strains crossed the placenta of principal gilts to infect fetuses in utero. The number of late-term dead fetuses (which appeared to be the best measure of relative virulence) ranged from 0 for litters of control gilts and gilts exposed to strain RespPRRS, to 38 for gilts exposed to strain NADC-8. All principal gilts became viremic and developed antibody against PRRSV. All strains persisted in alveolar macrophages of at least some principal gilts for at least 7 weeks after exposure. CONCLUSION: Strains of PRRSV vary in virulence. CLINICAL RELEVANCE: The effects of PRRSV on reproductive performance are strain dependent and this should be considered in making a tentative diagnosis on the basis of clinical observations. PMID- 8725810 TI - Liquid-phase blocking sandwich enzyme-linked immunosorbent assay for detection of antibodies against foot-and-mouth disease virus in water buffalo sera. AB - OBJECTIVE: To develop and apply the liquid-phase blocking sandwich ELISA (BLOCKING-ELISA) for the quantification of antibodies against foot-and-mouth disease virus (FMDV) strains O1 Campos, A24 Cruzeiro, and C3 Indaial. DESIGN- Antibody quantification. SAMPLE POPULATION: 158 water buffalo from various premises of Sao Paulo State-Brazil. The sera were collected either from systemically vaccinated or nonvaccinated animals. PROCEDURE: The basic reagents of BLOCKING-ELISA (capture and detector antibodies, virus antigens, and conjugate) were prepared and the reaction was optimized and standardized to quantify water buffalo antibodies against FMDV. An alternative procedure based on mathematical interpolation was adopted to estimate more precisely the antibody 50% competition titers in the BLOCKING-ELISA. These titers were compared with the virus-neutralization test (VNT) titers to determine the correlation between these techniques. The percentages of agreement, cutoff points, and reproducibility also were determined. RESULTS: The antibody titers obtained in the BLOCKING-ELISA had high positive correlation coefficients with VNT, reaching values of 0.90 for O1 Campos and C3 Indaial, and 0.82 for the A24 Cruzeiro (P < 0.0005). The cutoff points obtained by use of the copositivity and conegativity curves allowed determination of high levels of agreement between BLOCKING-ELISA and VNT antibody titers against the 3 FMDV strains analyzed. CONCLUSIONS: The results characterized by high correlation coefficients, levels of agreement, and reproducibility indicate that the BLOCKING-ELISA may replace the conventional VNT for detection and quantification of antibodies from water buffalo sera to FMDV. PMID- 8725811 TI - Effects of rate of gain on scrotal circumference and histopathologic features of the testes of half-sibling yearling beef bulls. AB - OBJECTIVE: To determine the effects of high and low gain on production variables and reproductive characteristics of bulls, including histopathologic lesions in the testes, as determined by quantitative measurements. ANIMALS: 23 half-sibling, 11.6-month-old beef bulls, averaging 326 +/- 8.3 kg in weight and having a mean scrotal circumference of 28.7 +/- 0.4 cm. DESIGN: Bulls were randomly assigned within pairs to high- or low-gain rations and fed ad libitum for 112 days. PROCEDURES: Scrotal circumference and body weight were measured on days 0, 28, 56, 84, and 112. Bulls were castrated on day 112. Morphologic characteristics of spermatozoa were determined from samples taken on days 50 and 111. Testis sections from each bull were examined, using light microscopy, for changes in the seminiferous epithelium and interstitium. A grading scheme was used to quantify testicular lesions by histologic techniques. Pathologic changes in 600 cross sections from each bull were classified into 9 grades. The degree of germinal epithelial loss per 100 tubules was calculated by assigning a value to each grade according to the severity of loss of germinal epithelium. RESULTS: Scrotal circumference (mean +/- SEM cm), 34.0 +/- 0.8 vs 31.7 +/- 0.5 (P < 0.001), body weight (kg), 512.3 +/- 19.6 vs 421.9 +/- 9.4 (P < 0.001), and paired testicular and epididymal weight (g), 532 +/- 31 vs 425 +/- 24 (P < 0.0001) and 73.8 +/- 1.9 vs 56.3 +/- 3.7 (P < 0.001) were greater in the high-gain than in the low-gain group. Morphologic characteristics of spermatozoa were similar for both groups. Means for graded seminiferous epithelial effects were similar in each group. Hypoplastic-type tubules were found in bulls with small scrotal circumference in both the high- and low-gain groups. CONCLUSION: High gain resulted in greater testicular growth but testicular structure and function were similar for both groups. PMID- 8725812 TI - Effect of Pasteurella multocida toxin on physeal growth in young pigs. AB - OBJECTIVE: To determine whether Pasteurella multocida toxin (PMT) affects growth of the proximal portion of the humerus of young pigs. ANIMALS: 20 colostrum deprived, cesarean-derived pigs. DESIGN AND PROCEDURE: 5 groups (n = 4/group) of pigs were formed. Group-1 pigs received 0.1 ml of phosphate-buffered saline solution for 4 weeks; group-2 pigs received 0.05 microgram of PMT/kg of body weight at 14 and 21 days; group-3 pigs received 0.05 microgram of PMT/kg at 28 and 35 days; group-4 pigs received 0.1 microgram of PMT/kg at 14 and 21 days; and group-5 pigs received hyperimmune serum (from a sow given purified toxin) on days 13, 20, 27, and 34, and 0.1 microgram of PMT/kg on days 14, 21, 28, and 35. RESULTS: All pigs given 0.1 microgram of PMT/kg without serum died or were euthanatized, as were 4 pigs given 0.05 microgram of PMT/kg. These pigs had increased serum interleukin 1 and 6 bioactivities. Pigs surviving 0.05 microgram of PMT had decreased weight gain, rough coat, marked atrophy of the ventral concha (as determined by turbinate perimeter ratios), and small stature. The surviving pigs also had reduced area and decreased proliferation indices in physeal chondrocytes on the basis of bromodeoxyuridine immunoreactivity. Control and serum-treated pigs gained weight, had no clinical effects, had similar physeal areas, and had higher cell proliferation indices. CONCLUSIONS: PMT inhibits endochondral bone formation by reducing physeal area and chondrocyte proliferation in vivo. Hyperimmune serum neutralizes the effects of toxin on weight gain, clinical appearance, physeal area, and chondrocyte proliferation. CLINICAL RELEVANCE: PMT may affect growth of the skeletal system. Antiserum to PMT is protective. PMID- 8725813 TI - Evaluation of antibodies reactive with porcine lymphocytes and lymphoma cells in formalin-fixed, paraffin-embedded, antigen-retrieved tissue sections. AB - OBJECTIVES: To determine whether antibodies raised against human or porcine lymphocytes are reactive with porcine lymphocytes and lymphoma cells in formalin fixed, paraffin-embedded, antigen-retrieved tissue sections, and to determine the conditions suitable for antigen retrieval. DESIGN: We evaluated reactivities of 27 anti-lymphocyte antibodies with porcine tissue sections, including those of lymphomas, treated with 11 antigen-retrieval methods. ANIMALS: Swine. PROCEDURE: We used 19 anti-human and 8 anti-porcine lymphocyte antibodies. For antigen retrieval, we tested 11 methods: heating with 6 soaking solutions in a microwave oven, heating with 2 commercially available soaking solutions in a water bath, and enzyme digestion with 3 proteases. After these treatments, sections were immunostained along with untreated sections. RESULTS: Of 27 anti-lymphocyte antibodies tested, CDw75 (clone LN-1), CD79 alpha (mb-1, clone HM57),CD79 beta (B29, clone B29/123), HLA-DR (clone TAL 1B5), and polyclonal CD3 were strongly reactive with porcine lymphoid tissues, including lymphomas, when sections were deparaffinized and heated in 0.1M Tris-HCl buffer (pH 8.0) in a microwave oven or with antigen-retrieval solution in a water bath. All patterns of positive staining were essentially the same as those previously described in human beings. CONCLUSION: These 5 antibodies, with the antigen retrieval sequences, are helpful in identifying, distinguishing, and characterizing lymphocyte subsets in archival sections of normal and pathologic porcine lymphoid tissues including lymphomas. In addition, our findings indicated that antigen retrieval by microwave heating of the sections can appreciably expand the range of antibodies useful in paraffin immunohistochemistry, including those that otherwise work only on sections from frozen tissues or from an antigen-derived species. PMID- 8725814 TI - Use of adverse conditions to stimulate a cellular stress response by equine articular chondrocytes. AB - OBJECTIVE: To determine the response of equine articular cartilage cells to heat and calcium stresses. DESIGN: Analysis of newly synthesized, [35S]methionine labeled proteins after treatment of isolated primary equine chondrocytes. PROCEDURE: Primary cultures of equine articular chondrocytes were incubated at temperatures ranging from 37 to 42 C for heat stress experiments or incubated in the presence or absence of the intracellular calcium pump inhibitor, thapsigargin, for calcium stress experiments. Patterns of new protein synthesis were determined by incubating with [35S]methionine followed by separation of proteins by use of one- or two-dimensional gel electrophoresis and visualization of labeled proteins by use of fluorography. RESULTS: Equine chondrocytes cultured at temperature of 42 C had increased synthesis of specific proteins, compared with the profile of protein synthesis in control chondrocytes cultured at 37 C. These changes were characteristic of the heat shock stress response described in a number of other mammalian cell-types. Equine chondrocytes cultured in the presence of thapsigargin also had increased synthesis of specific proteins. Two dimensional gel electrophoresis of these newly synthesized proteins revealed the changes to be consistent with the induction of the glucose-regulated protein family of stress proteins. CONCLUSIONS: Changes in the pattern of new protein synthesis can be induced in differentiated equine articular chondrocytes by heat shock or calcium stress. These responses are characteristic of a widely described mammalian stress response that has been postulated to be involved in cellular protective mechanisms. The ability of equine chondrocytes to mount a robust stress response may be important in the processes of tissue damage and recovery in articular joints of horses. PMID- 8725815 TI - Effects of pretreatment with ketoprofen and phenylbutazone on experimentally induced synovitis in horses. AB - OBJECTIVE: To compare the analgesic and anti-inflammatory effects of the nonsteroidal anti-inflammatory drugs (NSAID), ketoprofen (2.20 and 3.63 mg/kg of body weight) and phenylbutazone (4.40 mg/kg), in an acute equine synovitis model. DESIGN: 4 groups of 6 horses received NSAID or saline solution in a randomized design. ANIMALS: 24 clinically normal mares and geldings. PROCEDURE: Left intercarpal joints were injected with sterile carrageenan to induce synovitis at the same time as IV administration of NSAID or saline solution. Clinical assessments were made and synovial fluid was withdrawn at 0, 1, 3, 6, 9, 12, 24, and 48 hours. RESULTS: The eicosanoids, prostaglandin E2 (PGE2) and leukotriene B4, increased in synovial fluid after synovitis induction in all horses then returned to near baseline by 48 hours. All NSAID-treated horses had decreased PGE2, compared with saline-treated horses. This effect lasted longer in phenylbutazone-treated horses than in ketoprofen-treated horses. There were no treatment effects on leukotriene B4. In saline-treated animals, lameness, joint temperature, and synovial fluid volume, protein concentration, and nucleated cells increased 3 to 12 hours after induction, with marked reduction by 48 hours. Only phenylbutazone treatment reduced lameness, joint temperature, and synovial fluid volume. CONCLUSION: Phenylbutazone was more effective than ketoprofen in reducing lameness, joint temperature, synovial fluid volume, and synovial fluid PGE2. Results do not support lipoxygenase inhibition by either NSAID. CLINICAL RELEVANCE: This reversible model induced synovial fluid alterations similar to those observed in horses with septic arthritis. Results indicate that phenylbutazone may be more useful than ketoprofen in treating acute joint inflammation. PMID- 8725816 TI - Comparative efficacy of topically applied flurbiprofen, diclofenac, tolmetin, and suprofen for the treatment of experimentally induced blood-aqueous barrier disruption in dogs. AB - OBJECTIVE: To compare the relative efficacies of 4 topical nonsteroidal anti inflammatory drugs at preventing blood-aqueous barrier (BAB) disruption in dogs. DESIGN: 1 eye of each dog was treated with 1% suspensions of diclofenac, flurbiprofen, suprofen, or tolmetin, or with control solution. After 4 applications of eyedrops at 10-minute intervals, BAB disruption was induced in the treated eye by anterior chamber paracentesis. The severity of BAB disruption was measured by anterior chamber fluorophotometry. ANIMALS: 40 ocular-normal dogs. PROCEDURE: After pretreatment with eyedrops, rapid 100-microliters nonleaking anterior chamber paracentesis was performed in 1 eye of each dog to induce BAB disruption. 1 day after paracentesis, 1 ml of 10% fluorescein sodium was injected i.v.. The amount of fluorescein entering the anterior chamber of each eye was measured 30 to 60 minutes later by use of a computerized scanning fluorophotometer. The degree of BAB disruption was determined by comparing the amount of fluorescein entering the aqueous humor of the paracentesed eye with that of the nonparacentesed eye. RESULTS: At postparacentesis day 1, the order of statistically significant BAB-stabilizing efficacy among groups was: diclofenac > flurbiprofen > suprofen > tolmetin = control solution. CONCLUSIONS: Topically applied 1% suspensions of diclofenac, flurbiprofen, and suprofen are effective at preventing BAB disruption after paracentesis in dogs, indicating their potential usefulness for treatment of prostaglandin-mediated ocular disease. 1% tolmetin is no more effective than control solution. PMID- 8725817 TI - Apparent viscosity of the synovial fluid from mid-carpal, tibiotarsal, and distal interphalangeal joints of horses. AB - OBJECTIVE: To compare the apparent viscosity of normal synovial fluid of the mid carpal, tibiotarsal, and interphalangeal joints of horses. DESIGN: Viscosity evaluation over a range of shear rates was used to characterize the apparent viscosity of synovial fluids from the 3 joints. ANIMALS: 60 clinically normal adult horses. PROCEDURE: Viscosity data for synovial fluid samples were obtained over a shear rate range of 10 to 250/s and apparent viscosity was calculated at 50, 100, 150, 200, and 250/s. Effect of shear rate on apparent viscosity was determined, using a two-way ANOVA, with significant differences tested, using a Tukey's test at a significance level of P < 0.05. RESULTS: Synovial fluid from all these joints indicated shear thinning behavior: decreased apparent viscosity with increased shear rate. Apparent viscosity of synovial fluid from the 3 joints was not significantly different over the shear rate range of 50 to 250/s. CONCLUSION: Results of this study indicate that the apparent viscosity of the distal interphalangeal joint is not less than that of other joints, as has been reported. CLINICAL RELEVANCE: The observation of decreased synovial fluid viscosity of distal interphalangeal joint fluid should be considered as suggestive of joint disease. PMID- 8725819 TI - Effects of furosemide on hemorheologic alterations induced by incremental treadmill exercise in thoroughbreds. AB - OBJECTIVE: To determine whether furosemide treatment altered the blood flow properties and serum and RBC electrolyte concentrations of Thoroughbreds during submaximal treadmill exercise. DESIGN: Thoroughbreds were subjected to submaximal treadmill exercise with and without treatment with furosemide (1 mg/kg of body weight, IV). ANIMALS: 5 healthy Throughbreds that had raced within the past year and had no history of exercise-induced pulmonary hemorrhage. PROCEDURE: Venous blood samples were obtained before exercise, at treadmill speeds of 9 and 13 m/s, and 10 minutes after exercise, and hemorheologic and electrolyte test results were determined. RESULTS: Hemorheologic changes 60 minutes after furosemide administration included increased PCV, plasma total protein concentration, whole blood viscosity, mean RBC volume, and RBC potassium concentration, and decreased serum potassium concentration, serum chloride concentration, and RBC chloride concentration. Furosemide treatment attenuated the exercise-associated changes in RBC size, serum sodium concentration, serum potassium concentration, RBC potassium and chloride concentrations, and RBC density; exacerbated exercise associated increases in whole blood viscosity; and had no effect on RBC filterability. CONCLUSIONS: The hemorheologic effects of furosemide probably occurred secondary to total body and transmembrane fluid and electrolyte fluxes and would not improve blood flow properties. CLINICAL RELEVANCE: The beneficial effects of furosemide treatment in reducing the severity of bleeding in horses with exercise-induced pulmonary hemorrhage cannot be explained by improved blood flow properties. PMID- 8725818 TI - Influence of adrenergic and cholinergic mediators on the equine jejunum in vitro. AB - OBJECTIVE: To characterize the response of equine jejunal smooth muscle to adrenergic and cholinergic mediators. DESIGN: Evaluation of myogenic responses, using an in vitro model. SAMPLE POPULATION: Intestinal tissues were obtained from horses without gastrointestinal tract disorders or systemic disease. PROCEDURE: Baseline myogenic tone and amplitude and frequency of contraction were determined for suspended jejunal muscle strips. The level of adrenergic and cholinergic regulation was assessed, using atropine and adrenoceptor antagonists. The response of the muscles to norepinephrine was characterized, using adrenergic blockade and alpha- and beta-agonists. RESULTS: Adrenergic and cholinergic blockade had minimal effect on baseline myogenic activity. However, alpha 1- and beta 2-agonists induced significant (P < 0.05) decreases in the amplitude and frequency of contraction. Surprisingly, alpha 2-agonists caused an increase in the contraction amplitude of longitudinal muscle fibers (neurogenic in origin). Change in circular muscle activity was not induced by alpha 2-agonists. Norepinephrine induced a similar selective response and was inhibited by yohimbine. CONCLUSIONS: Baseline jejunal activity appears to be myogenic in origin and can function independently of sympathetic and parasympathetic input. However, intestinal smooth muscle can be affected by adrenergic agonists and potentially by increased concentrations of circulating catecholamines. Norepinephrine may act by altering the activity of other neurotransmitters. Differing responses between circular and longitudinal muscle fibers indicates a need to evaluate both components. CLINICAL RELEVANCE: Selective alpha 2-agonists may be potentially useful for motility modification of the equine jejunum. Therapeutic use of adrenergic blockade will be effective only in cases of increased adrenergic stimulation. PMID- 8725820 TI - Parasympathetic component of 5-hydroxytryptamine-induced pulmonary dysfunctions in healthy calves. AB - OBJECTIVE: To determine whether a cholinergic mechanism interferes with the pulmonary response to 5-hydroxytryptamine (5-HT) in the bovine species. DESIGN: The protocols differed with regard to the type of pretreatment calves were given 10 minutes before administration of 5-HT (0.05 mg/kg of body weight/min, over 2 minutes). Pretreatment consisted of saline, atropine, or hexamethonium solution given IV. ANIMALS: 6 healthy unsedated Friesian calves. PROCEDURE: Pulmonary function values were obtained before during, and after 5-HT infusions. RESULTS: After saline pretreatment, response to 5-HT consisted of immediate and brief apnea, bradycardia, and hypotension, followed by sustained tachypnea, tachycardia, pulmonary hypertension, and hypocapnic hypoxemia. Lung dynamic compliance (CLdyn) decreased to 19% of its baseline value, and total pulmonary resistance (RL) increased to 235%. Hexamethonium pretreatment resulted in a similar pattern of response except for the immediate and brief 5-HT-induced triad of apnea, bradycardia, and hypotension, which was suppressed. After atropine pretreatment, immediate and brief 5-HT-induced apnea-bradycardia-hypotension triad and sustained hypoxemia were abolished. In contrast, sustained tachypnea, tachycardia, pulmonary hypertension, and hypocapnia were maintained. Changes of CLdyn (59%) and RL (138%) were significantly attenuated. CONCLUSIONS: The initial and short-lasting response to 5-HT (ie. the apnea-bradycardia-hypotension triad) is mediated through a reflex central cholinergic pathway. The 5-HT-induced changes in CLdyn and RL are consistent with development of diffuse bronchoconstriction. Attenuation of these changes by atropine suggests that this bronchoconstrictor response to 5-HT is partly mediated through a cholinergic postganglionic pathway. PMID- 8725821 TI - Effect of milking frequency on milk somatic cell count characteristics and mammary secretory cell damage in cows. AB - OBJECTIVES: To examine whether once daily milking (ODM) increases somatic cell count (SCC) and causes changes in the proportion of macrophages, lymphocytes, or neutrophils in milk, and to examine whether ODM causes mammary secretory cell damage. DESIGN: During 3 successive periods, cows were milked twice daily (days 1 through 4), once daily (days 5 through 10), and twice daily (days 11 through 14); thus, each cow served as its own control. Milk samples were collected during each milking for SCC, bovine serum albumin (BSA; except days 1 and 14), and N-acetyl beta-D-glucosaminidase (NAGase) activity analyses. Differential cell counts were determined on days 2 (period 1), 5 and 8 (period 2), and 11 (period 3) in morning milk samples. ANIMALS: 8 multiparous Jersey cows. PROCEDURES: Cell counter (SCC), staining and counting (differential SCC), ELISA (BSA), and enzyme activity (NAGase). RESULTS: Once daily milking increased SCC (total and concentration), percentage of neutrophils, and BSA (content and total mass); during subsequent twice daily milking, SCC decreased, but BSA content and percentage of neutrophils remained increased. Activity of NAGase was not affected by ODM. CONCLUSIONS: ODM increases SCC, but this is not associated with damage to mammary secretory cells. The ODM-related sustained increase in BSA content coincided with an increase in neutrophil numbers, suggesting a causal relation. PMID- 8725822 TI - Vagal esophagomotor nerve function and esophageal motor performance in dogs with congenital idiopathic megaesophagus. AB - OBJECTIVES: To examine the integrity of the vagal efferent innervation to the esophagus and to assess esophageal motor performance in dogs with congenital idiopathic megaesophagus. DESIGN: An acute experimental protocol performed in control dogs and dogs with naturally acquired congenital idiopathic megaesophagus under pentobarbitone anesthesia. ANIMALS: 4 dogs with congenital idiopathic megaesophagus and 16 control dogs. PROCEDURE: Esophageal motor nerve conduction studies were performed by recording evoked compound motor action potentials from the tunica muscularis of the distal thoracic portion of the esophagus in response to supramaximal stimulation of the cervical portion of the vagus nerve at cranial and caudal sites. Subsequently, esophageal motor performance was measured over a wide range of esophageal muscle lengths by recording intraesophageal pressure responses to supramaximal twitch and tetanic stimulation of the cervical portion of the vagus at varying, stepwise amounts of esophageal distention. RESULTS: In dogs with congenital idiopathic megaesophagus, no electrophysiologic evidence was found for segmental demyelination or axonal degeneration in cervical vagal motor fibers innervating striated muscle of the thoracic esophagus portion. Nor was spontaneous EMG activity, indicative of esophageal muscle denervation or a primary myopathy, observed. In contrast, esophageal motor performance, which was dependent on esophageal dimensions, was reduced in dogs with congenital idiopathic megaesophagus. CONCLUSIONS: In dogs with congenital idiopathic megaesophagus, the vagal efferent innervation to the esophagus is likely to be normal, a primary esophageal myopathy is unlikely to be present, and the observed reduction in esophageal motor performance may arise as a secondary consequence of altered esophageal biomechanical properties rather than from a primary neuromuscular abnormality. PMID- 8725823 TI - Influence of methylprednisolone acetate on osteochondral healing in exercised tarsocrural joints of horses. AB - OBJECTIVE: To evaluate joint function and healing of surgically created full thickness articular cartilage defects in exercised horses after intra-articular administration of methylprednisolone acetate (MPA; 120 mg) and sterile saline solution in the contralateral limb. DESIGN: Experimental investigation. SAMPLE POPULATION: 12 healthy, sound, radiographically normal horses with induced full thickness osteochondral lesions on the medial and lateral trochlear ridges of the tali. PROCEDURE: Two 8.4-mm-diameter full-thickness articular cartilage lesions were created in each tarsocrural joint (12 horses [24 tarsocrural joints]); 1 was in a weight-bearing (WB) position and the other in a less weight-bearing (LWB) position. Each horse was maintained on a standardized exercise protocol (stall rest, days 0-6; walking, days 7-12; and treadmill, days 13-42) and evaluated throughout the study for changes in joint circumferences, synovial fluid, radiographs, lameness, and scintigraphy. 6 horses were euthanatized on day 42, and 6 on day 180. Gross morphometric assessment was performed, using an image analysis system on a projected color slide of the defect. The type of repair tissue, based on gross appearance, was expressed as a percentage of the total defect for each osteochondral defect. Histochemical assessment was performed, using safranin-O staining for proteoglycans and an image analysis system to express the area of stain uptake. Histomorphometric assessment was performed on H&E-stained sections, using an image analysis system. The repair tissue filling the defect was categorized as to tissue type and expressed as a percentage of the total defect area. Synovial membrane specimens were assessed semiquantitatively on H&E-stained sections for changes in character. Significance was established at P < 0.05. RESULTS: Joint circumference was significantly increased in the saline, compared with the MPA-treated, limbs on days 7, 12, and 42. Synovial fluid WBC counts were significantly increased in the MPA-treated limbs on day 42. Gross osteochondral defects had a greater percentage of mature repair tissue in saline treated joints (30.8% LWB, 23% WB), compared with MPA-treated joints (0% LWB, 0% WB) at 42 days Histomorphometric assessment of the repair tissue indicated significant differences with regard to the quality of repair in the saline treated (34% fibrous tissue LWB, 19.4% fibrous tissue WB) versus MPA-treated (2.5% fibrous tissue in LWB and WB) joints at 42 days. Microscopically, the percentage of fibrocartilage in the LWB (MPA, 23.7%; saline, 24.8%) was significantly greater than that in the WB (MPA, 14.6%; saline, 15.4%) site at day 180. The MPA-treated limbs had greater villous hyperplasia, edema, and extent of inflammation within the synovial membrane than did saline-treated limbs (days 42 and 180). CONCLUSION: MPA inhibits the development and maturation of repair tissue at 42 days and incites potential long-term (180 days) detrimental synovial membrane inflammation. Furthermore, a single dose of MPA does not cause long-term detrimental effects (180 days) in quality of repair-tissue. PMID- 8725824 TI - Laparoscopic anatomy of the abdomen in dorsally recumbent horses. AB - OBJECTIVES: To provide an accurate and detailed description of the laparoscopic anatomy of the abdomen of horses positioned in dorsal recumbency and to compare those observations with laparoscopic anatomy of standing horses. The effects of laparoscopy and positional changes on arterial blood pressure and blood gas values also were investigated. DESIGN: Descriptive anatomic study. SAMPLE POPULATION: Laparoscopy was performed on 6 horses (2 mares, 2 geldings, and 2 stallions) to record the normal laparoscopic anatomy of the abdomen in dorsal recumbency. PROCEDURE: Feed was withheld from all horses for 36 hours. Horses, under general anesthesia, were examined in horizontal and inclined positions (head-up and head-down). Intermittent positive-pressure ventilation was used, arterial blood pressure was continuously monitored, and samples for arterial blood gas measurements were taken at intervals. RESULTS: The main structures of diagnostic relevance observed in the caudal region of the abdomen were the urinary bladder, mesorchium and ductus deferens (left and right), left and right vaginal rings, insertion of the pre-pubic tendon, random segments of jejunum and descending colon, pelvic flexure of the ascending colon, body of the cecum, and cecocolic fold. The main structures observed in the cranial region of the abdomen were ventral surface of the diaphragm, falciform ligament and round ligaments of the liver, ventral portion of the left lateral, left medial, quadrate, and right lateral lobes of the liver, spleen, right and left ventral colons, sternal flexure of the ascending colon, apex of the cecum, and stomach. CONCLUSIONS: Alterations in cardiovascular and respiratory function in response to pneumoperitoneum and various positional changes indicated the need for continuous and thorough anesthetic monitoring and support. Comparison of anatomic observations made in dorsally recumbent, inclined horses with those reported for standing horses should enable practitioners to make patient positioning decisions that best suit access to specific visceral structures. Development of special instrumentation for manipulation of the viscera in horses, particularly the intestinal tract, would increase the diagnostic and therapeutic capabilities of laparoscopy during dorsal recumbency. PMID- 8725825 TI - Hyaluronate and large molecular weight proteoglycans in synovial fluid from horses with various arthritides. AB - OBJECTIVE: To investigate the presence of large molecular weight (MW) proteoglycans (PG) and hyaluronate (HA) in synovial fluid (SF) from horses with various arthritides and from control joints. DESIGN: Horses with acute (< 2 weeks) or chronic (> 4 weeks) lameness were examined by clinical examination, intrasynovial anesthesia, radiography, arthroscopy, and SF analysis. Joints were grouped on the basis of diagnosis: acute traumatic arthritis, chronic traumatic arthritis (with a subgroup of degenerative joint disease), intra-articular fracture, and infectious arthritis. ANIMALS: 31 horses with arthritis and 9 control horses; altogether 43 SF samples were analyzed. PROCEDURE: High performance liquid chromatography was used to assess HA and large MW PG in SF samples. RESULTS: A PG peak was identified in 8 of 23 SF samples of joints with chronic traumatic arthritis, 4 of which had no or minimal abnormal radiographic findings but mild articular cartilage fibrillation detected by arthroscopy, and in 3 joints with intra-articular fracture and 1 with resolving infectious arthritis, but not in joints with acute traumatic arthritis or in control joints. There was significant difference (P < 0.01) in mean (+/- SEM) HA concentration between control joints and joints with chronic traumatic arthritis (0.32 +/- 0.04 g/L; n = 9 vs 0.18 +/- 0.01 g/L; n = 23). CONCLUSION: Large MW PG fragments are released into equine SF in the course of articular disease and can be detected simultaneously with HA by high-performance liquid chromatography. CLINICAL RELEVANCE: The SF HA concentration can be used as diagnostic marker for chronic traumatic arthritis. However, SF PG or other marker cannot be used for diagnosing or monitoring degenerative joint disease. PMID- 8725826 TI - In vitro biomechanical comparison of the strength of the linea alba of the llama, using two suture patterns. AB - OBJECTIVE: To compare the strength of the sutured linea alba, in vitro, using 2 suture patterns. ANIMALS: 12 clinically normal llamas. PROCEDURE: 2 incisions in the linea alba of 12 llamas were closed with a simple continuous or inverted cruciate pattern, and tissue was harvested after 10 days. In 6 llamas the simple continuous line was intact; the inverted cruciate specimens contained 6 sutures. In 6 llamas, 1 knot was excised in the simple continuous pattern to simulate a failed line; the cruciate pattern contained 5 knots. Tissue sections were taken from cranial, between, and caudal to the linea alba incisions to compare fascial thickness. The sutured specimens were mounted in a mechanical testing system and tested to failure. A mixed-model ANOVA was used to evaluate the effects of suture pattern and incisional position on mechanical properties. RESULTS: Significant differences were not found between suture patterns or between location for yield force, failure force, or yield strain, whereas failure strain was lower for the intact simple continuous pattern than the inverted cruciate pattern (P = 0.003). From histomorphometric analysis, the caudal tissue specimens were significantly thinner than the middle tissue specimen cranial to the umbilicus (P = 0.006). CONCLUSION: There was no significant difference in monotonic breaking strength of the linea alba sutured with the simple continuous or inverted cruciate pattern. CLINICAL RELEVANCE: These results justify the use of the simple continuous pattern over the cruciate pattern for ventral midline closure in llamas because of the ease of placement and speed. PMID- 8725827 TI - Comparison of the degree of abdominal adhesion formation associated with chromic catgut and polypropylene suture materials. AB - OBJECTIVE: To evaluate the histologic pattern and biomechanical properties of adhesions caused by chromic catgut and polypropylene sutures, using an enteropexy model. DESIGN: Enteropexies were created in dogs, using chromic catgut and polypropylene suture. The adhesions associated with the enteropexies were examined histologically and mechanically. ANIMALS: 6 mixed-breed dogs weighing 16 to 20 kg. PROCEDURE: 72 enteropexies were created between the jejunum and abdominal wall. 36 sites were sutured with chromic catgut and 36 were sutured with polypropylene. 3 dogs were euthanatized after 1 week. The remaining dogs were euthanatized after 1 month. Samples of the enteropexy sites were obtained for histologic examination. The remaining sites were mechanically distracted until failure of the enteropexy site or adjacent tissue occurred. RESULTS: Histologic examination of the enteropexy sites did not reveal substantial differences in the degree of inflammation between the 2 suture types at 1 week or 1 month. The degree of inflammation decreased and the maturity of fibrous tissue formed at the enteropexy sites increased for all specimens over time. No statistically significant difference in breaking strength was observed between suture types at 1 week or 1 month. CONCLUSION: In dogs, the formation and strength of intentionally created abdominal adhesions are not increased by use of chromic catgut. CLINICAL RELEVANCE: Selection of chromic catgut suture for use in surgical procedures where adhesions are desired is unwarranted. PMID- 8725829 TI - Double impression procedure for removable partial denture retained with semiprecision attachments: a clinical report. PMID- 8725828 TI - Effects of dietary n-3 fatty acid supplementation versus thromboxane synthetase inhibition on gentamicin-induced nephrotoxicosis in healthy male dogs. AB - OBJECTIVE: To evaluate the protective effects of dietary n-3 fatty acid supplementation versus treatment with a thromboxane synthetase inhibitor (TXSI) in dogs given high-dose gentamicin. DESIGN: Clinicopathologic and renal histopathologic changes induced by gentamicin (10 mg/kg of body weight, IM, q 8 h, for 8 days) were compared in dogs fed an n-3 fatty acid-supplemented diet containing a fatty acid ratio of 5.7:1 (n-6:n-3), dogs treated with CGS 12970 (a specific TXSI given at 30 mg/kg, PO, q 8 h, beginning 2 days prior to gentamicin administration), and control dogs. The TXSI-treated and control dogs were fed a diet with a fatty acid ratio of 51.5:1 (n-6:n-3). Both diets were fed beginning 42 days prior to and during the 8-day course of gentamicin administration. ANIMALS: Eighteen 6-month-old male Beagles, 6 in each group. RESULTS: After 8 days of gentamicin administration, differences existed among groups. Compared with n-3-supplemented and control dogs. TXSI-treated dogs had higher creatinine clearance. Both TXSI-treated and n-3-supplemented dogs had higher urinary prostaglandin E2 and E3 (PGE2/3) and 6-keto prostaglandin F1a (PGF1a) excretion, compared with control dogs. Urinary thromboxane B2 (TXB2) excretion was higher in n-3-supplemented and control dogs, compared with TXSI-treated dogs. Urine PGE2/3 to-TXB2 and PGF(in)-to-TXB2, ratios were increased in TXSI-treated dogs, compared with n-3-supplemented and control dogs, and these ratios were increased in n-3 supplemented dogs, compared with control dogs. In addition, TXSI-treated and n-3 supplemented dogs had lower urinary protein excretion, compared with control dogs. Proximal tubular necrosis was less severe in TXSI-treated dogs, compared with control dogs. CONCLUSION: Treatment with CGS 12970 prior to and during gentamicin administration prevented increases in urinary TXB2 excretion and reduced nephrotoxicosis. CLINICAL RELEVANCE: Increased renal production/excretion of thromboxane is important in the pathogenesis of gentamicin-induced nephrotoxicosis. PMID- 8725830 TI - The contraindication of restoration to centric relation: a clinical report. PMID- 8725831 TI - Replacement of a fractured unilateral removable partial denture with a nonrigid fixed prosthesis: a clinical report. AB - Replacement of a fractured unilateral RPD with a five-unit nonrigid FPD was described in this clinical report, with the following advantages. (1) Swallowing or aspiration of a unilateral removable prosthesis will be avoided. (2) Fracture of solid metal connectors is diminished. (3) Metal-ceramic retainers and pontics will improve the patient's esthetic and functional demands. PMID- 8725832 TI - Repair of an eight-unit fixed partial denture with a resin-bonded overcasting: a clinical report. PMID- 8725833 TI - Application of implants in restorations in periodontal-prosthetic patients: a clinical report. PMID- 8725834 TI - Designs of bond strength tests for metal-ceramic complexes: review of the literature. AB - Despite the popularity of metal-ceramic restorations, the bonding mechanism remains the subject of theoretical discussion. Different experimental designs to evaluate bond strengths at the metal-ceramic interface are described. The effects of various stresses with each design are also discussed. A predictive experimental design should minimize variables and result in the least residual interfacial stresses. Dentists and dental technicians should benefit from comparisons of existing tests, and the discussion encourages introduction of more comprehensive approaches. PMID- 8725835 TI - Tensile bond strength of enamel-resin-metal joints. AB - This study observed the effects of the alloy type, resin type, and simulated occlusal loading on enamel-resin-metal tensile bond strength. Three different dental alloy samples were sandblasted with 50 microns of desiccant alumina and then bonded to the buccal and lingual halves of extracted human molars with four different dental resins. Specimens were divided into two groups, which included different halves of tooth samples. For one group tensile strength was tested after 19.8 N per 60,000 loading cycles. For the other group tensile testing was performed without cyclic loading. The results revealed that enamel-resin-metal bond strength was not affected by alloy type. However, a close relationship between the resin type and the bond strength was found. It was demonstrated that Panavia Ex and Concise resins produced greater bond strengths than did ABC and MBA cements. Also, a significant decrease in tensile bond strength values occurred after cyclic loading. PMID- 8725836 TI - Dimensional accuracy and stability of polymethyl methacrylate reinforced with metal wire or with continuous glass fiber. AB - The aim of this study was to determine the dimensional accuracy and stability of denture base polymethyl methacrylate (PMMA), which was reinforced in various ways. Autopolymerizing PMMA and heat-cured PMMA were reinforced either with semicircular steel wire or with a prefabricated experimental reinforcement made of continuous E-glass fiber. Control specimens had no reinforcement. The width of each U-shaped test specimen was measured with a digital micrometer under a light microscope immediately after the test specimen was cured and when stored in water for 1, 2, 7, and 14 days. The results revealed that both the type of PMMA and the type of reinforcement affected the dimensional accuracy of the test specimens, especially after 7 and 14 days in water storage (p < 0.001). The greatest dimensional accuracy was found with the unreinforced test specimen made from autopolymerizing PMMA and with test specimens reinforced with the metal wire made from heat-cured PMMA. In terms of the width of the test specimens, the lowest dimensional accuracy was found in unreinforced and glass fiber reinforced specimens made from heat-cured PMMA. Storage in water did not affect the stability of the width (p > 0.1). The results suggest that the polymerization shrinkage of PMMA causes lower dimensional accuracy of the test specimens reinforced with glass fiber. This should be considered when glass-fiber reinforcement is used clinically. PMID- 8725837 TI - Hidden clasps versus C clasps and I bars: a comparison of retention. AB - The Hidden clasp system overcomes the problem of poor esthetics of partial denture clasps, but it is not known how well it works as a retainer. This study tested the retention of Hidden clasps compared with that of C clasps and I bars when they function as part of a tooth-supported partial denture framework. Nine standardized frameworks that incorporated one of the three clasp designs on a premolar abutment were tested on a laboratory model. Their retention was recorded on a tensile testing machine. Specimens carrying the Hidden clasp had the greatest variability in retention among the three groups. Statistically significant differences were found between the C clasp and I bar groups. PMID- 8725838 TI - In vitro backscattering from implant materials during radiotherapy. AB - It is a certainty that many of the patients now receiving dental implants will one day require radiotherapy if they have head and neck cancer. The purpose of this study was to examine the dose enhancement form backscattered radiation at implant/bone interfaces irradiated with high-energy 6 MV and 10 MV x-rays. Three commonly used implant materials, pure titanium, Ti-6A1-4V alloy, and a high gold content implant material of various thicknesses, were used to measure the dose enhancement caused by backscattered radiation. The relative ionization charges at implant/bone interfaces were measured at distances of 0, 1, 2, and 3 mm away from the implant material by insertion of 0, 1, 2, and 3 mm thick bone substitute disks between the implant material and an ionization chamber. The results indicated that the highest dose enhancement occurred at a distance of 0 mm from the implant/bone interface for all the materials studied. The Au-Cu-Ag implant material had more average relative dose than pure titanium or Ti-6A1-4V alloy. The backscattered electrons decreased as the thickness of the bone substitute (distance between the implant material and the ionization chamber) increased. PMID- 8725839 TI - A conceptual framework for understanding outcomes of oral implant therapy. AB - The use of implants has expanded so rapidly that its effectiveness for many clinical situations has not been firmly established. Dentists are responsible for making appropriate therapeutic recommendations for the management of partial and complete edentulism. These decisions require an understanding of the consequences or outcome of treatment approaches. Outcome measures to evaluate the benefits of alternative therapies include longevity as well as physiologic, psychologic, and economic impacts. Future research should include outcome measures beyond implant prosthesis survival to more fully assess the practical impact of dental implants on the patient's oral health and to determine the most cost-effective approaches for managing completely and partially edentulous patients. PMID- 8725840 TI - Strength of porcelain fused to titanium beams. AB - The purposes of this study were to measure strengths of layered porcelain fused to titanium beams, determine failure modes, and investigate the porcelain titanium interface. A three-point flexural test and formulas derived especially for this purpose were used. The strength of layered porcelain-ceramic beams was limited by the cohesive tensile or compressive strengths of the porcelain, not by the porcelain-titanium interfacial bond, namely, the porcelain failure occurred at lower loads than did failure of the porcelain-titanium interface. The porcelain-titanium bond strength was at least 26 MPa. Scanning electron microscopy and energy-dispersive x-ray spectroscopy demonstrated that the bond was limited by delamination of a thin titanium-titanium oxide interface. Theoretic curves that describe effects of relative layer and total thickness on the force-bearing capacity of model beams were plotted. These curves indicated that porcelain-titanium prostheses should be made as thick as is practical, but the relative thickness of the porcelain and titanium layers would be less important. PMID- 8725841 TI - Lubrication of saliva substitutes at enamel-to-enamel contacts in an artificial mouth. AB - Mechanisms of salivary lubrication can be quantitatively measured by a reduction in the coefficient of friction. It is important that lubrication be assessed under the conditions of the oral cavity to properly assess lubrication regimes. The relative lubricity of three artificial salivas and two controls were assessed at a bovine enamel interface in an artificial mouth with a range of conditions that approximate oral function. Statistical analysis indicated that the enamel lubricity of sodium dodecylsulfate (SDS) and Oracare-D saliva substitutes were different from the other saliva substitutes and water. The low friction with Oracare-D and SDS saliva substitutes was because of resident amphipaths adsorbed at the enamel interface. Amphipaths adsorbed on enamel may provide a reduction in interocculsal friction and its resulting complications for patients with xerostomia. PMID- 8725842 TI - Dental education at the crossroads: a perspective from the chair. PMID- 8725843 TI - Dental education and practice in the 21st century: opportunities for excellence. AB - This study describes the environment that can be expected for dental education and dental practice as the 21st century approaches. The study uses data from many sources, including the recent report from the Institute of Medicine, which provides a comprehensive analysis of the variables that will affect dental education and practice in the next decade. Dental education and practice will be able to thrive if the new forces are carefully examined and the challenges are met promptly. PMID- 8725844 TI - The Dentist Scientist Award program and prosthodontics. AB - The Dentist Scientist Award (DSA) program was started by the National Institute of Dental Research (NIDR) in 1984 to prepare dentists for careers as clinical research scientists who would be highly skilled investigators and potential leaders in the full scope of oral health research. The DSA program provides support for dentists to undertake 5 years of intensive preparation in a course of study that includes basic and clinical science components integrated with a supervised research experience. From 1984 to 1994, the NIDR has invested +66.1 million dollars in the individual and institutional Dentist Scientist Award programs. Almost 250 people have been or are involved now in the DSA career development activity. Men comprise 72% of the recipients and women 28%. PMID- 8725845 TI - A graduate student's, graduate program director's, and chairman's perspective of the Dentist Scientist Award program. AB - The Dentist Scientist Award (DSA) program was introduced as an innovative mechanism of support and structure to meet the critical demand for the training of expert clinician scientists. This article presents an overview of the University of North Carolina's DSA program as it relates to the Department of Prosthodontics and the graduate program in prosthodontics. Recruitment of DSA candidates for the program and for faculty positions is described. PMID- 8725846 TI - Clinical evaluation in prosthodontics: practical methods to improve validity and reliability at the undergraduate level. AB - The potential causes of reliability and validity problems with traditional methods of evaluating a student's clinical prosthodontic skills are reviewed. Practical methods of evaluating critical skills in a reliable and valid way are presented. These methods include clinical tests using written criteria and anonymous examiners, skills assessment by faculty members, patient-based written examinations, and bell ringer examinations. PMID- 8725847 TI - A procedure for design of a posterior resin-bonded prosthesis. AB - A procedure is described that uses red and brown crayon-type pencils and graphite pencil lead with a surveyor to draw a color-coded design on a study cast before fabrication of a posterior resin-bonded prosthesis. During diagnosis and treatment planning, abutments on the study cast are analyzed for undercuts and the height of contour on each abutment tooth is marked with a graphite pencil lead. Metal retainers for the prosthesis are indicated with brown lines drawn on the abutments. Enamel surfaces requiring alteration to allow insertion of the prosthesis and provide resistance to displacement are marked in red on the study cast. The surveyed color-coded study cast can be used as a reference guide by both the clinician and the laboratory technician during fabrication of the posterior resin-bonded prosthesis. PMID- 8725848 TI - Veneer facing repair with a 4-META adhesive agent. PMID- 8725849 TI - Clinical study of location and reproducibility of three mandibular positions in relation to body posture and muscle function. PMID- 8725850 TI - Complete dentures for a child with hypohidrotic epidermal dysplasia: a clinical report. PMID- 8725851 TI - Does suppression of postprandial blood glucose excursions by the alpha glucosidase inhibitor miglitol improve insulin sensitivity in diet-treated type II diabetic patients? AB - OBJECTIVE: Insulin sensitivity is impaired in patients with type II diabetes and is exacerbated by high mean blood glucose (BG). Potentially, large postprandial swings in BG could result in further decrements of insulin sensitivity. Because alpha-glucosidase inhibitors cause a marked reduction in the amplitude of BG changes, the aim of this study was to determine if such a BG-smoothing effect improves insulin sensitivity in well-controlled type II diabetic subjects treated with diet alone. RESEARCH DESIGN AND METHODS: Patients received either miglitol (BAY m 1099) (50 mg three times daily) or placebo for 8 weeks in a randomized double-blind parallel study. The miglitol (9 men, 2 women) and placebo (7 men, 3 women) groups were well matched (mean +/- SD) for age, weight, and blood glucose control (fasting BG, 6.4 +/- 1.0 vs. 6.9 +/- 1.6 mmol/l; HbA1, 7.7 +/- 1.0 vs. 7.9 +/- 0.4%; fructosamine, 0.99 +/- 0.08 vs. 1.07 +/- 0.17 mmol/l). The glucose metabolic clearance rate was calculated during the last 30 min of a 150 min glucose/insulin sensitivity test (glucose, 6 mg . kg-1 . min-1; insulin, 0.5 U . kg-1 . min-1). RESULTS: There was no significant improvement in metabolic clearance rate (0.21 +/- 0.27 vs. 0.16 +/- 0.35 l . kg-1 . min-1) for the miglitol- and placebo-treated groups, respectively. There were no statistically significant differences between miglitol and placebo for changes from baseline in BG (0.1 +/- 0.1 vs. -0.1 +/- 0.2 mmol/l), HbA1 (0.1 +/- 0.1 vs. 0.3 +/- 0.1%), and fructosamine (-0.06 +/- 0.02 vs. -0.03 +/- 0.02 mmol/l). CONCLUSIONS: Alpha glucosidase-induced improvement in postprandial hyperglycemia does not result in increased insulin sensitivity. PMID- 8725852 TI - Erythrocyte Na(+)-K(+)-ATPase activity, metabolic control, and neuropathy in IDDM patients. AB - OBJECTIVE: To study the relationship among red blood cell Na(+)-K(+)-ATPase activity, metabolic control, and diabetic neuropathy. RESEARCH DESIGN AND METHODS: Na(+)-K(+)-ATPase activity has been measured in the red cell membrane of 43 long-standing IDDM patients (duration of diabetes 17.5 +/- 2 years, mean +/- SE), with 20 of the patients presenting with peripheral neuropathy. There were 23 healthy subjects serving as the control group. RESULTS: Na(+)-K(+)-ATPase activity was significantly lower in diabetic patients than in healthy subjects (236.5 +/- 7.5 vs. 294 +/- 10 nmol P1 . mg protein-1 . h-1, P < 0.05). Among diabetic patients, Na+/K(+)-ATPase activity was not dependent on the degree of diabetic control, nor was it correlated with either fasting blood glucose (r = 0.16, NS) or HbA1 (r = 0.01, NS). Na(+)-K(+)-ATPase activity was lower in patients with neuropathy than in those without it (212 +/- 8.5 vs. 261 +/- 6.6, P < 0.05). Furthermore, in a subgroup of 20 patients, a positive correlation was observed between erythrocyte Na(+)-K(+)-ATPase activity and nerve conduction velocity in the peroneal (r = 0.558, P < 0.02) and tibial nerve (r = 0.528, P < 0.05). CONCLUSIONS: These results suggest that diabetes-induced Na(+)-K(+)-ATPase activity dysfunction could be implicated in the pathogenesis of human diabetic neuropathy and the electrophysiological abnormalities observed in these patients. PMID- 8725853 TI - Increased resting and exercise-induced oxidative stress in young IDDM men. AB - OBJECTIVE: To assess the effect of acute physical exercise on oxidative stress and glutathione redox status and the relation to physical fitness in otherwise healthy young men with IDDM. RESEARCH DESIGN AND METHODS: Nine men with IDDM (HbA1 7.3 +/- 1.7%), ages 21-30 years, and 13 matched control subjects exercised on a bicycle ergometer for 40 min at 60% of their maximal oxygen consumption (VO2max). Oxidative stress was assessed with plasma thiobarbituric acid reactive substance (TBARS) levels (an index of lipid peroxidation) and, in response to exercise, also glutathione redox status. For glutathione redox status, blood total glutathione (TGSH) and oxidized glutathione (GSSG) were determined. Blood samples were drawn immediately before and after exercise. RESULTS: Resting plasma TBARS levels were markedly elevated in diabetic patients (2.2 +/- 0.7 vs. 0.9 +/- 0.4 mumol/l; P = 0.0002). Mean blood TGSH was higher in diabetic subjects (1,203 +/- 221 vs. 936 +/- 156 mmol/l; P = 0.002), with no significant difference in GSSG or GSSG/TGSH values. Exercise increased plasma TBARS and blood GSSG by approximately 50% in both groups. Resting plasma TBARS had a strong inverse correlation (r = -0.82; P = 0.006), and the exercise-induced percentage increase in TBARS had a strong positive correlation (r = 0.81, P = 0.008) with VO2max in diabetic subjects only. CONCLUSIONS: Glutathione redox status appears to be adequate in healthy young moderately active diabetic men. On the other hand, they demonstrated increased resting and postexercise oxidative stress as indicated by plasma TBARS. Although exercise acutely induces oxidative stress, in patients with diabetes, physical fitness may have a protective effect against oxidative stress. PMID- 8725854 TI - Glycemic responses to exercise in IDDM after simple and complex carbohydrate supplementation. AB - OBJECTIVE: Subjects with IDDM should take carbohydrate before exercise to avoid hypoglycemia. However, there is little information on the glycemic effect of recommended supplementation. This study is aimed to determine the glycemic effects of oral glucose or bread (30 g carbohydrate) before 45 min of moderate exercise. RESEARCH DESIGN AND METHODS: Nine subjects with uncomplicated IDDM did 45 min of bicycle ergometer exercise at 60% VO2max in the morning before insulin injection on three occasions: 1) with no carbohydrate supplement, 2) with 30 g glucose in water at -5 min, and 3) with 30 g carbohydrate as white bread with water at -20 min. The glycemic responses were determined. The glycemic responses to glucose and bread were also determined without exercise in six subjects. RESULTS: Without carbohydrate, exercise caused a small fall (-1.2 +/- 0.6 mmol/l, mean +/- SE) in plasma glucose (PG). With either glucose or bread, PG rose (the change in plasma glucose relative to basal [delta PG] = 5.1 +/- 0.8 and 2.6 +/- 0.8, respectively). The rise was greater (P < 0.01) without exercise (delta PG = 6.9 +/- 0.7 and 4.5 +/- 0.7, respectively). During exercise, glucose increased PG levels more than bread increased glucose levels P < 0.05). CONCLUSIONS: Before morning insulin injection, the fall in PG during moderate exercise in IDDM subjects is generally small or absent. The glycemic effects of complex carbohydrate are slightly less than glucose before exercise. Under these circumstances, the usually recommended amount of carbohydrate tends to cause an unwanted elevation of PG; thus, IDDM subjects should anticipate reducing or even omitting carbohydrate supplementation after monitoring their individual glycemic response. PMID- 8725855 TI - Glucagonostatic actions and reduction of fasting hyperglycemia by exogenous glucagon-like peptide I(7-36) amide in type I diabetic patients. AB - OBJECTIVE: Glucagon-like peptide I(7-36) amide (GLP-1) is a physiological incretin hormone that, in slightly supraphysiological doses, stimulates insulin secretion, lowers glucagon concentrations, and thereby normalizes elevated fasting plasma glucose concentrations in type II diabetic patients. It is not known whether GLP-1 has effects also in fasting type I diabetic patients. RESEARCH DESIGN AND METHODS: In 11 type I diabetic patients (HbA1c 9.1 +/- 2.1%; normal, 4.2-6.3%), fasting hyperglycemia was provoked by halving their usual evening NPH insulin dose. In random order on two occasions, 1.2 pmol . kg-1 . min 1 GLP-1 or placebo was infused intravenously in the morning (plasma glucose 13.7 +/- 0.9 mmol/l; plasma insulin 26 +/- 4 pmol/l). Glucose (glucose oxidase method), insulin, C-peptide, glucagon, GLP-1, cortisol, growth hormone (immunoassays), triglycerides, cholesterol, and nonesterified fatty acids (enzymatic tests) were measured. RESULTS: Glucagon was reduced from approximately 8 to 4 pmol/l, and plasma glucose was lowered from 13.4 +/- 1.0 to 10.0 +/- 1.2 mmol/l with GLP-1 administration (plasma concentrations approximately 100 pmol, P < 0.0001), but not with placebo (14.2 +/- 0.7 to 13.2 +/- 1.0). Transiently, C peptide was stimulated from basal 0.09 +/- 0.02 to 0.19 +/- 0.06 nmol/l by GLP-1 (P < 0.0001), but not by placebo (0.07 +/- 0.02 to 0.07 +/- 0.02). There was no significant effect on nonesterified fatty acids (P = 0.34), triglycerides (P = 0.57), cholesterol (P = 0.64), cortisol (P = 0.40), or growth hormone (P = 0.53). CONCLUSIONS: Therefore, exogenous GLP-1 is able to lower fasting glycemia also in type I diabetic patients, mainly by reducing glucagon concentrations. However, this alone is not sufficient to normalize fasting plasma glucose concentrations, as was previously observed in type II diabetic patients, in whom insulin secretion (C-peptide response) was stimulated 20-fold. PMID- 8725856 TI - Impaired glucose tolerance, diabetes, and cardiovascular disease risk factor profiles in the elderly. The Honolulu Heart Program. AB - OBJECTIVE: The relationship between glucose tolerance status and other cardiovascular disease (CVD) risk factors was evaluated in a cohort of Japanese American men (n = 3,741) ages 71-93 years who participated in the fourth examination of the Honolulu Heart Program in 1991-1993. RESEARCH DESIGN AND METHODS: In this cross-sectional study, subjects were classified by reported diabetes and glucose tolerance status using questionnaires and the World Health Organization (WHO) criteria, respectively. RESULTS: The prevalence of reported diabetes was 17%. Among the men who completed an oral glucose tolerance test and had no history of diabetes (n = 1,900), 23% were diagnosed as diabetic and 39% had impaired glucose tolerance (IGT) by WHO criteria. The CVD risk factor profiles of men with IGT and diabetes were significantly more adverse compared with men with normal glucose tolerance after adjustment for age. The rates of hypertension, mean levels of BMI, waist-to-hip ratio, triglycerides, and fasting insulin were higher in men with IGT and diabetes compared with normal subjects. Opposite trends were observed for HDL cholesterol. Two-hour insulin was significantly higher among men with IGT and previously undiagnosed diabetes. Men with known diabetes had a lower physical activity index and higher fibrinogen levels than normal subjects. No significant differences were observed for current smoking and alcohol intake. Differences in risk factor levels by glucose tolerance status remained after adjustment for age, physical activity, BMI, and waist-to-hip ratio. CONCLUSIONS: These findings show that among elderly men of Japanese ancestry, impaired glucose tolerance and undiagnosed and known diabetes are highly prevalent, and these conditions are associated with adverse CVD factor profiles. PMID- 8725857 TI - Glucose and amino acid turnover in untreated gestational diabetes. AB - OBJECTIVE: Although gestational diabetes affects as many as 3% of all pregnant women, specific aspects of glucose and protein metabolism in this population have not been clearly delineated. We tested the hypothesis that gestational diabetes mellitus (GDM) results in increased glucose production and proteolysis during fasting. RESEARCH DESIGN AND METHODS: Using tracer isotope infusions, the rate of appearance (Ra) of glucose, leucine, phenylalanine and tyrosine, phenylalanine hydroxylation, leucine oxidation, and urea nitrogen excretion were determined after an overnight fast in 10 GDM subjects, within 2 weeks of diagnosis and before initiation of treatment, and in a matched control group of nine healthy nondiabetic pregnant women. RESULTS: Fasting glucose Ra was similar in GDM patients and control subjects (GDM, 12.8 +/- 1.1 vs. control subjects, 12.8 +/- 0.9 mumol . kg-1 . min-1). Leucine and phenylalanine Ra (reflecting proteolysis) also were not different between GDM patients and control subjects (GDM leucine Ra, 128 +/- 14 vs. control subjects, 124 +/- 5; phenylalanine Ra GDM, 35 +/- 4 vs. control subjects, 40 +/- 2 mumol . kg-1 . h-1). Furthermore, leucine oxidation and phenylalanine hydroxylation were not increased in GDM subjects, urea nitrogen excretion was actually lower in GDM patients. However, fasting insulin concentrations were significantly elevated in GDM subjects (GDM, 165 +/- 35 vs. control subjects, 30 +/- 5 pmol/l; P < 0.01). CONCLUSIONS: Hepatic glucose release and whole-body proteolysis in GDM patients were remarkably similar to matched pregnant control subjects. This was achieved with insulin concentrations three- to fivefold higher than normal, suggesting significant insulin resistance for both glucose and protein metabolism in GDM. PMID- 8725858 TI - Low risk of post-cesarean section infection in insulin-requiring diabetic women. AB - OBJECTIVE: The purpose of this study was to determine if insulin-requiring diabetic women undergoing nonelective cesarean section are at higher risk for postoperative infection than nondiabetic women. RESEARCH DESIGN AND METHODS: Medical records of a cohort of insulin-requiring diabetic women who underwent cesarean section after labor or rupture of membranes and nondiabetic control subjects matched for age and insurance status were retrospectively reviewed. Data abstracted included maternal characteristics, antepartum, intrapartum, and postpartum events. RESULTS: Post-cesarean section infection including endometritis, wound infection, and septic pelvic thrombophlebitis occurred in 10.2% of 205 diabetic women and 12.1% of control subjects, in whom antibiotic prophylaxis was used in 79% of diabetic women and 84% of control subjects. Duration of rupture of membranes was a significant risk factor for post-cesarean section infection in both groups. CONCLUSIONS: Insulin-requiring diabetic women undergoing nonelective cesarean section with antimicrobial prophylaxis have a rate of postoperative infection similar to that for nondiabetic women. PMID- 8725859 TI - Quality of outpatient care provided to diabetic patients. A health maintenance organization experience. AB - OBJECTIVE: To document the quality of diabetes care provided to patients in a large health maintenance organization (HMO) from 1 January 1993 to 1 January 1994 and compare it to the standards of the American Diabetes Association (ADA). RESEARCH DESIGN AND METHODS: To meet a Health Plan and Employer Data Information Set (HEDIS) requirement, a major HMO in California identified 14,539 members with diabetes and randomly selected 384 individuals for review. Charts were available on 353 of these patients, and after obtaining the information for the HEDIS review, additional information was extracted from the charts by an outside chart reviewer. This data set was used for an analysis of the quality of diabetic care provided by the participating medical groups to these HMO members during 1 year. Documentation of follow-up and measures of glycemic and lipid control was examined both for absolute values and for the frequency of measurement over the year. These results were compared to the ADA standards of care. RESULTS: Although patients averaged 4.5 visits to their primary care physicians (PCPs) over the year, 21% had one or fewer visits per year. Glycated hemoglobin levels were not documented in 56% of patients (ADA recommends two to four measurements per year), and of those with a glycated hemoglobin level measured. 39% had at least one value > or = 10%. Fasting plasma glucose concentrations were not documented in 65% of patients (four to six per year recommended). Foot exams (which should be performed at each regular visit) were not documented for 94% of patients. Urine protein measurements were not performed in 52% of patients. Additionally, many patients had elevated and untreated lipid abnormalities. CONCLUSIONS: In spite of the frequency of PCP visits during the year for many of these patients, diabetes management was inadequate. This lack of adequate preventive care will lead to an increased risk of the development of the acute and chronic complications of diabetes, creating an even greater future burden on the health care system and negative consequences for patients. PMID- 8725860 TI - Risk factors predicting lower extremity amputations in patients with NIDDM. AB - OBJECTIVE: To examine the predictors of lower extremity amputation in patients with NIDDM. RESEARCH DESIGN AND METHODS: At baseline, risk factors for amputation were determined in 1,044 NIDDM patients (571 men, 473 women) aged 45 to 64 years. These patients were followed up to 7 years with respect to amputation. RESULTS: The incidence of amputation was 5.6% in men and 5.3% in women. High fasting plasma glucose at baseline examination and the duration of diabetes were associated with a twofold risk for amputation. Similarly, glycemic control measured at baseline by HbA1 was an important predictor of amputation. There was a dose-response relationship between plasma glucose or HbA1 and the risk for amputation. The effect of hyperglycemia on the risk of amputation was seen clearly even after the adjustment for other cardiovascular risk factors. Signs of peripheral neuropathy and bilateral absence of Achilles tendon reflexes and vibration sense were important predictors for amputation. Furthermore, absent peripheral artery pulses and femoral artery bruit on auscultation predicted amputation. CONCLUSIONS: Our 7-year follow-up study gives strong evidence that poor glycemic control is an important predictor of amputation in patients with NIDDM in addition to clinically detectable peripheral arterial disease and peripheral neuropathy. PMID- 8725861 TI - Promoting weight loss in type II diabetes. AB - OBJECTIVE: To examine strategies-behavioral therapies, exercise, diet, anorectic drugs, surgery, or a combination of strategies-used for promoting weight loss in people with type II diabetes. RESEARCH DESIGN AND METHODS: Meta-analysis was used to synthesize research of promoting weight loss in the population. Literature search strategies involved reviewing bibliographies, conducting computer searches and surveys of relevant master's degree programs, and contacting representatives of the Centers for Disease Control. The final sample consisted of 89 studies involving 1,800 subjects. Data were extracted on 80 variables characterizing the sample of studies/subjects and on 23 outcome variables, including weight, metabolic control, lipids, and other physiological parameters. RESULTS: Diet alone had the largest statistically significant impact on weight loss (-20 lb) and metabolic control (-2.7% in glycosylated hemoglobin). All diets significantly improved fasting blood sugar. Behavioral programs alone had a statistically significant impact on weight loss (-6.4 lb) and metabolic control (-1.5%) but effects were less than for diet alone. Data from the few exercise studies indicated that weighted average effects for exercise on weight loss (-3.4 lb) and metabolic control (-0.8%) were less than diet alone. Behavioral therapy plus diet plus exercise was associated with statistically significant effect size estimates for weight loss (-8.5 lb) and metabolic control (-1.6%). Diet alone achieved better results. Effects of weight promotion strategies, in general, were smaller in experimental studies and for individuals over age 55. CONCLUSIONS: Dietary strategies are most effective for promoting short-term weight loss in type II diabetes. A number of gaps exist in the extant literature- descriptions of subjects, interventions, or longitudinal outcomes beyond 12 months after intervention. PMID- 8725862 TI - Influence of smoking on the survival rate of diabetic patients requiring hemodialysis. AB - OBJECTIVE: To study the influence of cigarette smoking on the survival rate of diabetic patients on hemodialysis. RESEARCH DESIGN AND METHODS: We evaluated 1- and 5-year survival rates and the mean values of HbA1c, serum lipids, fibrinogen, and blood pressure. We compared the prevalence of vascular damage at the beginning of dialysis therapy and the endpoint of the study, as well as the causes of death in 22 diabetic patients who smoked (> 10 cigarettes/day) and 30 nonsmoking diabetic patients. RESULTS: There were no differences with respect to HbA1c, cholesterol, and triglycerides. In contrast, diabetic patients with tobacco consumption had significantly (P < 0.05) higher levels of fibrinogen (428 +/- 98 vs. 378 +/- 76 mg/dl) and higher systolic blood pressures (154 +/- 12 vs. 146 +/- 13 mm Hg) than the nonsmoking group. The 1- and 5-year survival rates of the smoking patients were 68 and 9%, respectively, and in the nonsmoking subjects, 80 and 37%, respectively (P < 0.05). The prevalence of vascular damage was similar in both groups, but at the endpoint of the study, the incidence of myocardial infarctions was significantly higher (P < 0.005) in the smoking patients (77 vs. 13%). Cardiovascular events were the most frequent cause of death in both patient groups but more frequently in the smoking subjects (80 vs. 63%). CONCLUSIONS: Hemodialyzed diabetic cigarette smokers show higher fibrinogen and systolic blood pressure values, a higher incidence of myocardial infarctions, and their 5-year survival rate is significantly decreased when compared with nonsmoking patients on hemodialysis. PMID- 8725863 TI - The dense LDL phenotype. Association with plasma lipoprotein levels, visceral obesity, and hyperinsulinemia in men. AB - OBJECTIVE: To investigate the potential relationship between the cluster of metabolic abnormalities found in visceral obesity and the small dense LDL phenotype. RESEARCH DESIGN AND METHODS: We have estimated LDL peak particle size by nondenaturing 2-16% gradient gel electrophoresis in a sample of 79 men. Glucose tolerance and fasting plasma insulin and lipoprotein levels were also measured. RESULTS: The LDL particle score, calculated from migration, distances and relative band intensities and reflecting the proportion of small dense LDL particles, was positively correlated with plasma triglyceride (TG) (r = 0.60, P < 0.0001) and negatively correlated with HDL cholesterol (r = -0.56, P < 0.0001) levels. Although the LDL particle score was not associated with variations in plasma LDL cholesterol or LDL apolipoprotein (apo) B concentrations, it was significantly correlated with the LDL apo B-to-LDL cholesterol ratio (r = 0.60, P < 0.0001). Fasting plasma insulin and visceral adipose tissue (AT) areas measured by computed tomography were weakly but significantly correlated with the LDL particle score (r = 0.23 and 0.29, respectively, P < 0.05). LDL peak particle size showed similar but inverse correlations with anthropometric and metabolic variables. Subjects classified as having small dense LDL particles (by comparing subjects in the highest tertile versus those in the lowest tertile of the LDL particle score distribution) were characterized by increased plasma TG, reduced HDL cholesterol, higher fasting insulin levels, and elevated visceral AT accumulation. However, multiple regression analyses revealed that visceral AT accumulation was not an independent predictor of the dense LDL phenotype after inclusion of TG and HDL cholesterol levels and lipoprotein ratios in the model. CONCLUSIONS: It thus appears that the high TG-low HDL cholesterol dyslipidemia frequently found in visceral obesity and in a hyperinsulinemic state is a strong correlate of the small dense LDL phenotype. Although associated with the dense LDL phenotype, visceral obesity and hyperinsulinemia were not independent predictors of an increased proportion of small dense LDL particles after controlling for TG and HDL cholesterol levels. PMID- 8725864 TI - Diabetic foot infections. Bacteriology and activity of 10 oral antimicrobial agents against bacteria isolated from consecutive cases. AB - OBJECTIVE: To study the relative frequency of bacterial isolates cultured from community-acquired foot infections and assess their comparative in vitro susceptibility to sparfloxacin, levofloxacin, and eight other commonly used oral antimicrobial agents. RESEARCH DESIGN AND METHODS: This is a prospective study in which the infected wounds of 25 consecutive diabetic patients seen by one of the authors were cultured as they entered the hospital. Isolates were stored and tested for susceptibility to 10 oral antimicrobial agents using the agar dilution method. RESULTS: Staphylococcus aurcus was the most common isolate (76% of patients), including methicillin-resistant S. aurcus (MRSA) in 5 of 25 (20%) patient wounds. Streptococci, enterococci, Enterobacteriaceae, and anaerobes were also present in > or = 40% of patient wounds. Sparfloxacin and levofloxacin were the most active agents tested with activity against > or = 88% of isolates. Isolates resistant to sparfloxacin and levofloxacin included MRSA, enterococci, and some anaerobes. When analyzed by prior exposure to antibiotics, patients who had previously received oral antibiotics were more likely to have MRSA, enterococci, and Pseudomonas aeruginosa isolated and less likely to have Enterobacteriaceae and anaerobes isolated from their wounds. CONCLUSIONS: MRSA and enterococci are now a common cause of diabetic foot infections, and the increased prevalence may be due to antimicrobial use. These wounds may require use of combined antimicrobial therapy for initial outpatient management. The new fluoroquinolones, sparfloxacin and levofloxacin, were the most active oral agents tested. PMID- 8725865 TI - An importance of carbohydrate ingestion for the expression of the effect of alpha glucosidase inhibitor in NIDDM. AB - OBJECTIVE: To examine the usefulness of alpha-glucosidase inhibitors in glycemic control of patients with NIDDM. The involvement of carbohydrate ingestion in manifestation of the effects of alpha-glucosidase inhibitors was also investigated. RESEARCH DESIGN AND METHODS: A total of 41 patients hospitalized with NIDDM (22 patients receiving sulfonylurea and 19 receiving insulin therapy) were given alpha-glucosidase inhibitors during the period when their blood glucose levels were well controlled. They were followed for 3 weeks as inpatients and for an additional 6 months as outpatients. They were retrospectively divided into two groups according to the percentage of carbohydrates in all sources of calories during outpatient management: the < 50% group and the > 50% group. Between these two groups, we compared circadian variation in blood glucose levels, HbA1c, and urine C-peptide. RESULTS: Treatment with alpha-glucosidase inhibitors during the hospital stay markedly improved circadian variation in blood glucose levels and HbA1c and decreased urine C-peptide in both groups. While HbA1c returned to its pretreatment level at 6 months after the treatment in the < 50% group, HbA1c had further improved in the > 50% group at 6 months. CONCLUSIONS: alpha-Glucosidase inhibitors are useful for glycemic control in patients with NIDDM and the percentage of carbohydrate in all calorie sources is an important factor for the expression of their effects. PMID- 8725866 TI - Changing behavior. Practical lessons from the diabetes control and complications trial. PMID- 8725867 TI - Effect of follow-up of women with gestational diabetes on the ratio of IDDM to NIDDM in pregnancy. AB - OBJECTIVE: We wished to test the hypothesis that the diagnosis of diabetes in women with previous gestational diabetes in our follow-up program had altered the ratio of IDDM to NIDDM in our pregnant population. RESEARCH DESIGN AND METHODS: We identified all pregnancies managed at the Mercy Hospital for Women in Melbourne, Australia, from 1971 to 1994 that were complicated by prepregnancy diabetes. In these 374 pregnancies, we identified those women who had previously been diagnosed with gestational diabetes mellitus (GDM). The changing prevalences over time of prepregnancy IDDM and NIDDM, as well as the contribution to both of these conditions made by women who had previously had GDM, were calculated. RESULTS: Over the period of the study, there was an increase in the prevalence of IDDM from 0.15 to 0.44% (chi 2 for trend, P < 0.00001) and NIDDM from 0.03 to 0.11% (chi 2 for trend, P = 0.0001). The proportion of all women with diabetes with NIDDM did not change significantly (16.7-20%). There was a progressive increase in the proportion of women with NIDDM who had had GDM (from 8.3 to 39.1%), but the trend was not statistically significant (P = 0.059). Women with NIDDM were more likely (20 of 64, 31.3%) to have had gestational diabetes in the past than women with IDDM (12 of 310, 3.9%, odds ratio 11.3, 95% CI 5.16-24.7, P < 0.0001). CONCLUSIONS: Despite finding relatively young women to have NIDDM, our GDM follow-up clinic has not yet altered significantly the ratio of IDDM to NIDDM in pregnancy. PMID- 8725868 TI - Effect of long-term glycemic control on cognitive function. AB - OBJECTIVE: To investigate the relationship between recurrent hypoglycemia and cognitive impairment in insulin-dependent diabetic patients. RESEARCH DESIGN AND METHODS: Seventy patients who were diagnosed as diabetic at age 18 years or older, were under 55 years old, and had no condition likely to affect cognitive abilities were recruited from a diabetic register. Patients were interviewed to obtain information on the frequency of major and minor hypoglycemia. Their cognitive abilities were assessed on tests of premorbid intelligence, current intelligence, reaction time, concentration, memory, and information processing. RESULTS: There was a significant correlation between the apparent decline in intelligence, expressed as the discrepancy between the estimated premorbid and the actual performance intelligence quotient, and the frequency of major hypoglycemic attacks (rs = -0.30; P < 0.01). Comparison of patients with and without recurrent hypoglycemia showed few significant differences in cognitive ability. CONCLUSIONS: Results support previous work that suggests that major hypoglycemic attacks have a significant effect on some aspects of cognitive function, but the clinical importance of this finding remains to be determined. PMID- 8725869 TI - Proinsulin secretion during the first 3 years after diagnosis in diabetic patients with and without islet cell antibodies. AB - OBJECTIVE: To evaluate proinsulin secretion in different types of NIDDM. RESEARCH DESIGN AND METHODS: Proinsulin and insulin were evaluated at diagnosis of diabetes and 3 years later (fasting and after stimulation with intravenous glucose and glucagon) in 10 NIDDM patients without islet cell antibodies (ICAs) at diagnosis (age 52 +/- 4 years), 11 NIDDM patients with ICAs at diagnosis (age 50 +/- 5 years), and 21 healthy control subjects (age 53 +/- 4 years). RESULTS: At diagnosis, fasting proinsulin was higher in NIDDM patients without ICAs than in control subjects (39.6 +/- 10.0 vs. 12.8 +/- 1.6 pmol/l, P < 0.01). Proinsulin response to intravenous glucose decreased in NIDDM patients with ICAs (from 35.6 +/- 6.2 to 13.5 +/- 5.4 pmol/l, P < 0.05), but remained unchanged in those without ICAs. At 3 years after diagnosis, fasting proinsulin (10.0 +/- 3.7 vs. 59.1 +/- 17.0 pmol/l) and proinsulin responses to intravenous glucose (13.5 +/- 5.4 vs. 103.9 +/- 35.1 pmol/l) and to intravenous glucagon (7.4 +/- 3.9 vs. 36.0 +/- 7.7 pmol/l) were much lower (P < 0.01) in NIDDM patients with ICAs than in those without ICAs. CONCLUSIONS: After diagnosis of diabetes, proinsulin secretion decreases significantly in NIDDM patients with ICAs and remains constant in those without. PMID- 8725870 TI - The prevalence of diabetes in Rio de Janeiro, Brazil. The Cooperative Group for the Study of Diabetes Prevalence in Rio De Janeiro. AB - OBJECTIVE: To assess the prevalence of diabetes and impaired glucose tolerance (IGT) in the adult population of Rio de Janeiro, a two-stage cross-sectional survey was carried out in a random sample of 2,051 individuals aged 30-69 years from Rio de Janeiro city in Brazil. RESEARCH DESIGN AND METHODS: Subjects were first screened by fasting capillary glycemia (FCG). All individuals who screened positive (FCG > 5.6 mmol/l) and every sixth consecutive person who screened negative (FCG < 5.6 mmol/l) were subjected to a 75-g glucose load. Diagnoses of diabetes and IGT were based on World Health Organization criteria. RESULTS: Results from every sixth individual who screened negative were extrapolated to all individuals who screened negative after adjustment for some potential bias in the subsample. Age-adjusted prevalence rates for diabetes and IGT were 7.1 and 9.0%, respectively. The rates were higher (P < 0.01) among women than among men (8.7 vs. 5.2% for diabetes and 11.7 vs. 5.8% for IGT), among obese individuals than among nonobese individuals (7.9 vs. 6.2% for diabetes and 11.4 vs. 7.3% for IGT), and among those with family history of diabetes than among those without family history of diabetes (12.4 vs. 4.8% for diabetes and 13.8 vs. 6.7% for IGT). The rates for diabetes and IGT increased with age, being 1.7 and 4.5%, respectively, for the age-group of 30-39 years, 3.9 and 8.5% for the age-group of 40-49 years, 13.6 and 13% for the age-group of 50-59 years, and 17.3 and 15.3% for the age-group of 60-69 years (P < 0.01). The prevalence of diabetes was higher among individuals with low educational levels than among those with high educational levels (7.3 vs 4.2%). For IGT, the rates increased from the group with intermediate level of education (8.3%) to the low- (11.3%) and high education group (12.6%). Differences in the rates for whites and non-whites (6.9 vs. 7.1% for diabetes and 8.8 vs. 9.6% for IGT) were not statistically significant. Among those with confirmed diabetes in the survey, 27.6% did not know of their diabetic condition. Among previously diagnosed diabetes (self reported diabetes), 19.5% were not being treated, 31.8% were on diet only, 40.7% were on oral hypoglycemic drugs, and 8.0% were on insulin. Self-reported prevalence of diabetes was 0.1% for the population < 30 years of age, 4.3% for the 30-69 year old age-group, and 16.6% for those > 70 years of age. CONCLUSIONS: The numbers found for Rio de Janeiro are similar to those for more developed countries and lead us to conclude that the impact of diabetes on public health is the same as in those countries where this disease is considered an important health problem. PMID- 8725871 TI - Bromocriptine (Ergoset) reduces body weight and improves glucose tolerance in obese subjects. AB - OBJECTIVE: A double-blind placebo controlled study investigated long-term effects of Ergoset, a new quick release formulation of bromocriptine, on body weight, body fat, and glucose tolerance in a group (n = 17) of obese subjects who were instructed to follow a moderate hypocaloric diet. RESEARCH DESIGN AND METHODS: Obese individuals (> 25% body fat for men and > 30% body fat for women) were instructed to follow a calorie-restricted diet (70% of weight maintaining based on study entry weight) and were randomized to daily treatment with Ergoset (1.6 2.4 mg/day) or placebo at 0800 over an 18-week treatment period. Oral glucose tolerance tests were performed on subjects before initiation and again at termination of treatment. Body weight and body fat (determined by skinfold measurements) were quantified every 2 weeks during the course of treatment. RESULTS: Ergoset treatment for 18 weeks significantly reduced body weight and body fat versus placebo (6.3 +/- 1.5 and 5.4 +/- 1.1 kg vs. 0.9 +/- 1.0 and 1.5 +/- 0.6 kg. respectively, P < 0.01). Ergoset, but not placebo, also improved glucose tolerance (P < 0.02); the stimulated area under the oral glucose tolerance curve was reduced by 46% (from 121 +/- 23 to 64 +/- 32 mg.h-1.dl-1), while the stimulated area under the insulin curve was reduced by 30%. CONCLUSIONS: When combined with instruction to follow a moderate hypocaloric diet, Ergoset, but not placebo, improves glucose tolerance and promotes significant weight and body fat loss in obese subjects over an 18- week treatment period. PMID- 8725872 TI - Diabetic ketoacidosis in young obese Japanese men. PMID- 8725873 TI - Metformin useful in combination with exogenous insulin. PMID- 8725874 TI - Increased prevalence of mitral valve prolapse in IDDM. PMID- 8725875 TI - Spurious ketonuria due to captopril and other free sulfhydryl drugs. PMID- 8725876 TI - GAD antibodies in IDDM in Thailand. PMID- 8725877 TI - Transduction of human renal carcinoma cells with human gamma-interferon gene via retroviral vector. AB - We used a retroviral vector containing a human gamma-interferon (gamma-IFN) gene to transduce 13 renal carcinoma cell lines. The transduction efficiencies ranged from 0% to 60%, as determined by using an analogous vector containing the LacZ marker gene. In addition, gene-transferred resistance to the antibiotic neomycin was used to select for transduced cells. Nine of 13 lines were successfully transduced. Transduction was associated with the morphologic change of elongation, and there was a marked decrease in cell growth rate. Transduced cells secreted varying amounts (20-1076 pg/10(6) cells/d) of gamma-IFN as measured by enzyme-linked immunosorbent assay for at least 2 to 3 weeks after transduction (including 1 day of transduction, 6-7 days of selection, and an additional 8-12 days before the first passage of the transduced cells). Human leukocyte antigen (HLA) class II expression was markedly increased in six of seven cell lines; HLA class I expression was significantly increased in two of eight lines. Transduced cells that were subjected to cryopreservation after irradiation still produced gamma-IFN and expressed HLA class I and II antigens, although generally at lower levels than before these manipulations. This study confirms that retroviral vector transduction of the human gamma-IFN gene into renal carcinoma cells is feasible and associated with persistent production of gamma-IFN and increased expression of HLA class I and II molecules, and these effects are retained after irradiation and cryopreservation. This suggests that an autologous tumor cell vaccine trial with irradiated gamma-IFN gene-transduced renal carcinoma cell is rationale and feasible. PMID- 8725878 TI - Potential salmon sperm origin of the E3 region insert of the adenovirus 5 dl309 mutant. AB - The plasmid pJM17 is a commonly used adenoviral backbone derived from the dl309 mutant virus. It contains unknown sequences inserted in the E3 region during construction of the dl309 mutant. Complete description of the backbone sequence is required for interpretation of potential vector effects and for regulatory approval of a vector to be used in clinical trials. The anonymous E3 insert was sequenced and analyzed. The insert fragment is 646 base pairs (bp) long and is 100 bp shorter than the vector sequences it replaces. It interrupts the expression of the E3B 10.4K, 14.6K, and 14.7K genes, but not the E3A glycoprotein (gp) 19K gene. Sequence analysis and Southern blotting suggest that the insert might originate from salmon sperm DNA used as carrier during the construction of dl309. Transcription from the insert was not detected by Northern blot analysis of vector-transduced cells but was detected by reverse transcriptase polymerase chain reaction. PMID- 8725879 TI - Escherichia coli gpt gene sensitizes rat glioma cells to killing by 6 thioxanthine or 6-thioguanine. AB - Genes that encode enzymes that convert inactive "prodrugs" into anticancer metabolites may be therapeutically useful against brain tumors. Unlike other genes tested to date in brain tumor models, the Escherichia coli gpt gene is unique in that it not only sensitizes cells to the prodrug 6-thioxanthine (6TX) but also encodes resistance to a different regimen (mycophenolic acid, xanthine, and hypoxanthine), thus providing a means to select for gpt-positive cells. In the present study, rat C6 glioma cells were infected with a retrovirus vector that transduces this gene. A clonal line (C6GPT-7) was derived that exhibited significant 6TX susceptibility in vitro with an ID50 of 2.5 mumol/L, whereas 50% growth inhibition of parental C6 cells was not achieved at concentrations tested (up to 50 mumol/L). This line also exhibited significant sensitivity to 6 thioguanine (6TG), with an ID50 of 0.05 mumol/L, whereas 50% growth inhibition of parental C6 cells was achieved at 0.5 mumol/L. In a "bystander" assay, C6GPT-7 tumor cells efficiently transferred 6TX sensitivity to C6 cells at ratios as low as 1:9 (C6GPT-7:C6). This in vitro bystander effect was abrogated when C6GPT-7 and C6 cells were separated by a microporous membrane, suggesting that it was not mediated by highly diffusible metabolites. In vivo both 6TX and 6TG significantly inhibited the growth of subcutaneously transplanted C6GPT-7 cells but not that of C6 cells in athymic mice. In an intracerebral model, both 6TX and 6TG exhibited significant antiproliferative effects against tumors formed by C6GPT-7 cells. These findings provide a basis for exploring further gene therapy strategies based on in vivo transfer of the E coli gpt gene to provide chemosensitivity against 6TX and 6TG. PMID- 8725880 TI - Recombinant vaccinia expressing interleukin-2 for cancer gene therapy. AB - Use of a recombinant vaccinia virus expressing human interleukin-2 (IL-2) was evaluated for preparation of tumor vaccines. A/J mice were immunized against neuroblastoma (C1300) cells using a preparation of C1300 cells infected/transfected with the recombinant virus, vCF13, expressing IL-2. A second recombinant vaccinia, vSC8, expressing Escherichia coli beta-galactosidase, was used as a control. After three weekly immunizations with virus-transfected cells, the mice were challenged with 1 x 10(6) unmodified C1300 cells and tumor development was monitored. Tumor development in the mice was inhibited by immunization with vCF13-transfected cells, compared to those vaccinated with vSC8 transfected cells (P < .008). A group of mice (7/15) immunized with vCF13 transfected cells followed by tumor challenge survived more than 60 days, at which time all mice immunized with the control vaccine were dead (p < .006). Five of the mice treated with the vCF13 vaccine were alive for more than 75 days (P < .05), after which they were rechallenged with another dose of 1 x 10(6) unmodified tumor cells. Tumor development was not apparent in these mice for more than 45 days following the second challenge, suggesting that these mice were completely protected by this immunization. These results demonstrate that recombinant vaccinia virus expressing IL-2 may be useful for cancer gene therapy. PMID- 8725881 TI - Mutant SV40 large T antigen as a therapeutic agent for HER-2/neu-overexpressing ovarian cancer. AB - The HER-2/neu gene is frequently amplified and/or its protein product, p185, is overexpressed in a number of human cancers. Overexpression of p185 correlates with poor prognosis and low survival rates in ovarian cancer patients. We previously found that the K1 mutant of SV40 large T antigen inhibits rat neu promoter and suppresses mutation-activated rat neu transformation in mouse fibroblasts. We show here that K1 also inhibits human HER-2/neu promoter in human ovarian cancer cells. To investigate whether K1 can suppress HER-2/neu transformation and thus is a potential therapeutic agent, we used an orthotopic ovarian cancer model in which mice were injected intraperitoneally with HER-2/neu overexpressing human ovarian cancer cells to induce tumor development. The tumor bearing mice were then treated with K1-liposome complex weekly. We found that liposome-mediated K1 gene transfer decreased the p185 protein level by K1 expression in these cancer cells and significantly prolonged mice survival; about 40% of these treated mice were alive for more than 1 year without any tumor development. On the other hand, the animals from control groups that did not receive this gene therapy all developed tumors and died within 7 months. The results indicate that liposome-mediated K1 gene transfer is able to suppress tumor development from HER-2/neu-overexpressing ovarian cancer cells in mice. PMID- 8725882 TI - Plasmid DNA gene therapy: studies with the human interleukin-2 gene in tumor cells in vitro and in the murine B16 melanoma model in vivo. AB - The plasmid DNA vector pVCL-1102 containing the coding sequence for the human IL 2 gene was evaluated for expression in tumor cells in vitro and in vivo. In vitro transfection of murine B16 tumor cells with pVCL-1102 resulted in the expression of 36,000 IU (5.7 micrograms) of biologically active IL-2/10(6) cells/48 h. In vitro transfection of human tumor lines and primary cultures from human biopsies with pVCL-1102 resulted in the expression of 1,289 to 9345 IU of IL-2/10(6) cells/48 h and 30 to 794 IU of IL-2/10(6) cells/48 h, respectively. In vivo, direct intratumor injection of pVCL-1102 resulted in retention of intact plasmid DNA in the tumor tissue and IL-2 secretion by cell cultures derived from the injected tumors. Formulation of pVCL-1102 with the cationic lipid DMRIE/DOPE inhibited DNA degradation and enhanced in vivo transfection efficiency over plasmid DNA alone. Antitumor activity of the pVCL-1102/DMRIE/DOPE complex was evaluated in a B16 melanoma model in mice. An IL-2-specific effect could not be demonstrated in a subcutaneous model because the intratumor injection of plasmid DNA lacking the IL-2 coding sequence also resulted in a significant reduction in tumor volume. However, an IL-2-specific effect was observed when B16 cells were transfected in vitro prior to implantation into the mouse. Transient transfection of B16 cells with pVCL-1102 rendered the cells less tumorigenic in vivo and produced a significant reduction in tumor volume. These data demonstrate that a plasmid DNA expression vector can be used to deliver the IL-2 gene to tumor cells in vitro and in vivo, resulting in the expression of significant levels of IL-2 protein. These data also illustrate the need for the use of appropriate controls when evaluating the in vivo biological activity of plasmid DNA in murine tumor models. PMID- 8725884 TI - Dual expression of human leukocyte antigen molecules and the B7-1 costimulatory molecule (CD80) on human melanoma cells after particle-mediated gene transfer. AB - The aim of this study was to determine if human melanoma cells could be molecularly modified by particle-mediated gene transfer with a "gene gun", using genes for interferon-gamma (IFN-gamma), the B7-1 costimulatory molecule (CD80), and human leukocyte antigen (HLA)-A2, to augment expression of both HLA molecules and B7-1. Established and early passage melanoma cells transfected with human IFN gamma complementary DNA (cDNA) produced IFN-gamma (50-5,000 pg/mL). The biological effect of this IFN-gamma transgene included an upregulation, or de novo appearance, of HLA expression. These melanoma cells had no detectable baseline surface expression of the B7-1 costimulatory molecule, but 8% to 31% of these cells became B7-1 positive with no selection procedure after gene transfer with human B7-1 cDNA. After combination gene transfer with cDNAs for both IFN gamma and B7-1, 9% to 33% of these cells expressed both HLA-DR and B7-1. In combination gene transfer experiments with cDNAs for both HLA-A2 and B7-1, dual expression of HLA-A2 and B7-1 was achieved in 10% to 17% of the melanoma cells. Thus, the molecular modification of human melanoma cells to increase expression of both HLA and B7-1 can be achieved by particle-mediated gene delivery and presents a promising strategy to stimulate antimelanoma T-cell immunity. Key words: Melanoma; T cells; B7-1 costimulatory molecule (CD80); major histocompatibility complex. PMID- 8725883 TI - In vivo particle-mediated cytokine gene transfer into canine oral mucosa and epidermis. AB - Cytokines can stimulate immune effector cells present within the oral mucosa and epidermis to respond to vaccination or to combat cancer. However, intravenous cytokine delivery is often inefficient and frequently accompanied by systemic toxicity. The goal of this study was to evaluate dogs as a large animal model for gene therapy of cancer because they develop spontaneous oral and epidermal tumors. In this report, we demonstrate that particle-mediated gene transfer of beta-galactosidase, luciferase, interleukin-2, interleukin-6, and granulocyte macrophage colony stimulating factor (GM-CSF) complementary DNA (cDNA) into the oral mucosa and epidermis of healthy dogs resulted in effective, localized, transgenic protein expression. Additionally, the epidermal sites transfected with GM-CSF developed a profound inflammatory reaction characterized by neutrophilic infiltration. Clinical pathology analyses were unremarkable. These results demonstrate that in vivo particle-mediated gene transfer of canine oral mucosa and epidermis with cytokine cDNA can result in production of biologically active transgenic cytokines with minimal toxicity. These findings have applications to cancer immunotherapy using a gene gun approach. PMID- 8725885 TI - Novel costimulators in the immune gene therapy of cancer. AB - One of the major goals of cancer immunotherapy is the induction of tumour specific T-lymphocyte responses that will be effective in the rejection of established tumours. The prospects for such therapy rely on the identification of tumour antigens, and although there is persuasive evidence for the presence of such antigens,1,2 the occurrence of the disease does illustrate that the immune system is at least, on some occasions, unable to recognise and destroy these targets. Tumour antigens may be novel proteins (from genetic lesions or viral infections), modified existing antigens (eg, abnormally glycosylated cell surface proteins), or inappropriately expressed normal gene products (eg, CA125, carcinoembryonic antigen, and alpha-fetoprotein).1 Involvement of the immune system in the normal surveillance and suppression of cancer is further suggested by the increased incidence of tumours in immunocompromised patients.3 However, recent evidence has shown that, at least in model systems, cancer cells can be modulated in such a way that they stimulate cells of the immune system to recognise and destroy these malignant cells. This review summarizes the costimulatory molecules involved in the activation of such cells, the principles and mechanisms underlying their activation, and how such knowledge can be used to persuade the immune system to challenge cancer. PMID- 8725886 TI - Screening for prostate cancer: opportunities for prevention. AB - Although early detection and treatment of prostate cancer is widely advocated, this so-called secondary prevention approach has a number of drawbacks. First, it is not yet certain that active treatment of localized prostate cancer offers any advantage over surveillance. Second, screening may detect indolent tumors while missing some virulent ones. Third, treatment is not uniformly successful, even in patients with early disease. Fourth, radical prostatectomy and radiotherapy are associated with considerable side effects. And finally, the economic and psychological costs of large-scale screening cannot be overlooked. Although attention has been focused on the possibility of primary prevention, neither large-scale dietary manipulation nor long-term prophylactic use of retinoids is considered feasible. With the recent approval of the 5-alpha-reductase inhibitor finasteride for the treatment of benign prostatic hyperplasia, the opportunity for primary chemoprevention has moved closer to reality. The Prostate Cancer Prevention Trial (PCPT), a randomized, placebo-controlled study expected to enroll 18,000 healthy men over the age of 55, is currently addressing the question of whether finasteride prophylaxis can reduce the incidence of prostate cancer over a 7-year period. This is a US government work. There are no restrictions on its use. PMID- 8725887 TI - Risk factors for progression in patients with prostate cancer treated with radical prostatectomy. AB - Adjuvant therapy after radical prostatectomy should ideally be limited to those patients at greatest risk for cancer recurrence, but identification of these patients remains a challenge. Although tumor volume has traditionally been regarded as the most important prognostic factor in patients with localized prostate cancer, a recent multivariate analysis has shown that tumor is not an independent predictor. Moreover, accurate measurement of tumor volume is extremely difficult. Preoperative serum prostate-specific antigen (PSA) levels have been identified as a significant independent predictor of progression. Disease-free survival is significantly better in patients with DNA-diploid prostate cancers than in those with nondiploid tumors. Histological grade is also a powerful predictor of disease progression. As a basis for selecting candidates for adjuvant therapy, clinical staging is too inaccurate and pathological staging too subjective. A recent Mayo Clinic study assessed the value of widely available clinical and laboratory parameters in predicting treatment failure after radical prostatectomy in 904 patients with pathologically organ-confined prostate cancer. Multivariate analysis identified Gleason score, preoperative serum PSA concentration, and DNA ploidy as independent predictors for progression. These risk factors were used to develop a scoring system that allows patients to be classified according to their risk of progression. Patients in the highest risk categories might be targeted for adjuvant therapy and closer surveillance, whereas those at lower risk might be followed less frequently. PMID- 8725888 TI - Histopathological effects of androgen deprivation in prostatic cancer. AB - Thanks to earlier detection of clinically significant prostatic adenocarcinoma by measurement of serum prostate-specific antigen (PSA) levels, increasing numbers of patients are undergoing radical prostatectomy. However, the curative potential of this procedure is seriously limited by clinical understaging, which results in positive surgical margins and a marked increase in disease progression. In a multicenter study, histopathologic evaluation of radical prostatectomy specimens showed that presurgical androgen deprivation with leuprolide plus flutamide reduced the incidence of surgical margin involvement by 62%. In patients who received androgen deprivation therapy, characteristic and recognizable histopathologic changes in nontumor glands included atrophy, basal cell prominence, vacuolated luminal cell layers, and squamous and transitional cell metaplasia. Androgen deprivation markedly reduced the incidence of prostatic intraepithelial neoplasia (PIN) to 35%. The effects of androgen deprivation on prostatic carcinoma included smaller tumor glands, pyknosis and empty glandular spaces, and vacuolization and degeneration of tumor cells with an inflammatory response. Similar but less pronounced changes with no decrease in PIN were observed in finasteride-treated patients. It is important for pathologists to be aware of these histological changes and process tissue appropriately, because the changes affect the recognition and histological grading of tumors in radical prostatectomy specimens. PMID- 8725889 TI - Neoadjuvant total androgen suppression and radiotherapy in the management of locally advanced prostate cancer. AB - The combination of radiation therapy and total androgen suppression shows potential for improving the outcome in patients with locally advanced prostate cancer. Both radiation and androgen ablation induce apoptosis through different mechanisms, and a synergistic interaction has been reported between the two modalities in vitro. Studies of neoadjuvant hormonal therapy before radical prostatectomy have shown considerable tumor shrinkage, a change that would greatly facilitate subsequent radiotherapy. A recent randomized trial involving patients with bulky disease has shown that the use of androgen ablation therapy before and during radiation therapy substantially improved local control and decreased the time to biochemical failure. Still another randomized trial performed in patients with locally advanced disease has shown that the positive biopsy rate was strikingly lower in patients who received neoadjuvant hormonal therapy before radiation than in those who underwent radiotherapy alone, and was even lower in patients who were treated with both neoadjuvant and adjuvant total androgen suppression. The combination of hormonal therapy and radiation has not yet been proven to prolong survival. Nevertheless, given the limitations of conventional radiotherapy, it is recommended that patients at high risk for failure be entered into one of the ongoing Radiation Therapy Oncology Group (RTOG) trials. These trials are attempting to clarify such questions as the role of adjuvant therapy following neoadjuvant therapy and radiation, the optimal timing of hormonal therapy, and the role of whole pelvic irradiation. PMID- 8725890 TI - Optimal duration of neoadjuvant androgen withdrawal therapy before radical prostatectomy in clinically confined prostate cancer. AB - Experimental studies have shown that neoadjuvant androgen therapy dramatically reduces the rate of local recurrence after tumor excision. In the clinical setting, a 3-month course of neoadjuvant therapy before radical prostatectomy has been shown to significantly reduce positive margin rates, but follow-up is too short to assess the impact of such therapy on biochemical and clinical recurrence rates. A phase II study using an ultrasensitive assay showed that 8 months of neoadjuvant therapy were required before prostate-specific antigen (PSA) levels to reach their nadir in 84% of study participants. The positive margin rate in this study was substantially lower than those reported in the literature. Importantly, restaging of specimens after prostatic acid phosphatase (PAP) immunostaining did not upstage or increase positive margin rates. In addition, prolonged neoadjuvant therapy did not appear to result in progression of androgen independent clones. A randomized phase III trial has been initiated to determine whether an 8-month course of neoadjuvant hormonal therapy is superior to a 3 month course in reducing positive margin rates and biochemical recurrences in patients with clinically confined prostate cancer. PMID- 8725891 TI - Neoadjuvant hormonal treatment before radical prostatectomy. AB - Randomized trials evaluating the use of androgen deprivation therapy (ADT) before radical prostatectomy have consistently shown significant decreases in prostate volume and in serum prostate-specific antigen (PSA) levels. All but one study have reported that hormonal pretreatment significantly reduces the incidence of positive surgical margins. However, androgen deprivation does not significantly influence seminal vesicle extension or lymph node involvement nor does it affect tumor grade. The fibrotic reactions that sometimes result from hormonal pretreatment can increase the difficulty of surgery but this has not been associated with a higher incidence of perioperative or postoperative complications. In closely monitored clinical trials, bothersome side effects of neoadjuvant hormonal therapy have been limited. Issues that remain to be addressed are the optimal duration of neoadjuvant treatment and whether the benefits of such therapy will translate into improved disease-free survival. At this time, patients with clinical stage T2b disease, PSA elevations greater than 10 to 20 ng/mL, and a high Gleason grade may be considered candidates for neoadjuvant hormonal treatment before surgery. PMID- 8725892 TI - Premature aging in Werner's syndrome spares the central nervous system. AB - Werner's Syndrome is a rare genetic disease, characterized by premature aging of many tissues and organs. We studied the brain morphology and function in two patients with Werner's syndrome to assess the possible involvement of the central nervous system in this premature aging process. The two patients (brother and sister, respectively) were studied by magnetic resonance imaging (MRI) and angiography (MRA), single photon emission computed tomography (SPECT) with (99mTc)-d,l-hexamethyl propilene amine oxime (HMPAO), positron emission tomography (PET) with 2(18F)-Fluoro-2-deoxyglucose (FDG), electroencephalography (EEG), and electromyography (EMG). Some of these investigations were also repeated after 1 year. The results of all these studies were normal. The premature aging process in patients with Werner's syndrome, while affecting most tissues, seems to spare the central nervous system. PMID- 8725893 TI - Neurofibrillary tangle-associated alteration of stathmin in Alzheimer's disease. AB - Stathmin (p19), a 19-kDa cytosolic phosphorotein, plays a key role in converting extracellular signals into intracellular biochemical changes. Antibodies and cDNA specific for stathmin were used to study its levels and localization in normal and Alzheimer's disease (AD) brain tissue. The stathmin protein concentration was reduced in AD neocortex as assessed by Western blotting, whereas the concentration of its mRNA detected by both in situ hybridization and slot blot were increased in AD. The alteration of the stathmin protein concentration was negatively correlated with neurofibrillary tangle numbers but not with plaque numbers. Immunoreactivity was evenly localized to the cytoplasm of neurons in control cortical sections, whereas in AD it was preferentially localized to some of the neurofibrillary tangle-bearing neurons. Numbers of stathmin-positive neurons were inversely correlated with tangle numbers but not with plaque numbers in the frontal cortex of AD patients. PMID- 8725894 TI - Glycogen synthase kinase 3 alpha and 3 beta do not colocalize with neurofibrillary tangles. AB - Glycogen synthase kinase (GSK) 3 alpha and 3 beta are two proline-directed serine/ threonine kinases that have been shown in vitro to hyperphosphorylate tau, and therefore, may contribute to neurofibillary tangle (NFT) formation in Alzheimer's disease (AD). We report here that, in the human hippocampal formation of both control and AD individuals, GSK 3 alpha and 3 beta are immunohistochemically localized to neurons within the presubiculum > CA1, CA3, and CA4 subfields of the hippocampus, layers III > II > IV, V, VI of entorhinal cortex, and occasional neurons in layers III, V, and VI of temporal neocortex. By contrast, NFTs occur primarily in CA1. subiculum, layers II and IV of entorhinal cortex, and layers II, III, and V of temporal neocortex. The presubiculum and other subfields are frequently spared. Thus, localization of GSK 3 alpha and GSK 3 beta does not correspond to the expected pattern of neuronal vulnerability to NFT formation in AD. Interpreted within the limitations of immunohistochemical detection, these results argue against a major role of GSK 3 alpha or GSK 3 beta in NFT formation in AD. PMID- 8725895 TI - Quantitative 1H and 31P MRS of PCA extracts of postmortem Alzheimer's disease brain. AB - Several previous studies have shown metabolic abnormalities in perchloric acid extracts of postmortem Alzheimer's disease (AD) brain by both proton (1H) and phosphorus-31 (31P) magnetic resonance spectroscopy (MRS). In all of these studies the results were expressed in relative terms, in units of mol percent. The results of this study, expressed in the absolute units of mumol/g wet weight, verify the previous 1H and 31P MRS studies. Absolute increases were found for myo inositol, aspartate, L-glutamate, alanine, phosphocholine, and the phosphodiesters,. Absolute decreases were found for phosphoethanolamine and N acetyl-l-aspartate. Many of these changes also were observed in non-AD dementia brain extracts, but changes in myo-inositol, inositol-l-phosphate, aspartate, and L-glutamate appeared to be more specific for AD in extracts of many brain areas. These results suggest that compounds related to membrane degradation and excitatory neuro-transmission increase in Alzheimer's disease while compounds related to neuronal integrity and inhibitory neurotransmission are decreased. PMID- 8725897 TI - Neuropathology in controls and demented subjects from the Baltimore Longitudinal Study of Aging. AB - To establish correlations among cognitive states and neuropathology, we have examined 22 subjects (69-97 years of age) from the Baltimore Longitudinal Study of Aging (BLSA), of whom 15 had normal and stable cognitive performances and seven had dementia of variable severity. In the majority of normal subjects, few or no beta-amyloid (A beta) deposits or senile plaques (SP) were present in the neocortex, but neurofibrillary tangles (NFT) were consistently found in CA1 of hippocampus and layer II of entorhinal cortex. In two (15%) normal individuals, the densities of SP were consistent with the diagnosis of possible Alzheimer's disease (AD). We speculate that these cases with normal cognitive states and abundant neocortical SP may represent preclinical AD. We conclude that the neocortex of a majority of cognitively intact individuals can remain free of A beta deposits or SP, even into the tenth decade of life. PMID- 8725896 TI - Human brain S100 beta and S100 beta mRNA expression increases with age: pathogenic implications for Alzheimer's disease. AB - S100 beta is a neurite extension factor that has been implicated in the development of neuritic plaques in Alzheimer's disease. We analyzed the expression of S100 beta and its encoding mRNA, using immunohistochemistry, enzyme linked immunosorbent assay, and Northern blot analysis, in postmortem brain tissue from 26 neurologically normal patients, aged 1-80 years. Tissue levels of S100 beta and S100 beta mRNA, as well as the number of S100 beta-immunoreactive (S100 beta +) astrocytes, increased with advancing age (r = 0.60, p = 0.008; r = 0.65, p = 0.007: and r = 0.73, p = 0.001, respectively). In patients more than 60 years old, the number of S100 beta + astrocytes and the tissue levels of S100 beta and S100 beta mRNA were significantly higher than those in patients less than 60 years of age (p = 0.001, p = 0.035, and p = 0.047, respectively). All of these values, however, were significantly less than those found in Alzheimer patients (p < 0.05 or better). Our findings, together with the known functions of S100 beta, suggest that age-related increases in S100 beta expression are important in the pathogenesis of Alzheimer's disease and may explain in part the increased incidence of this disease with advancing age. PMID- 8725898 TI - APOE alleles in Alzheimer's disease and vascular dementia in a population aged 85+. AB - Apolipoprotein E genotyping was carried out in a stratified random sample of 52 patients with Alzheimer's disease, 48 patients with vascular or mixed dementia, and 49 nondemented controls in a population-based study of people aged 85 and older (the Vantaa 85+ Study). Our results indicate that the apolipoprotein E epsilon 4 allele is associated with approximately a twofold increase in clinically diagnosed Alzheimer's disease in this very old general population aged 85+. When combined with previous studies, our data also suggest that the association is decreasing with age. In contrast, there appears to be no relation between apolipoprotein E alleles and clinically diagnosed vascular dementia. PMID- 8725899 TI - Retinal pathology in Alzheimer's disease. I. Ganglion cell loss in foveal/parafoveal retina. AB - Morphometric analysis of the numbers of neurons in the ganglion cell layer (GCL) of the central retina (fovea/foveola/parafoveal retina) in eyes from 9 Alzheimer's disease (AD) and 11 age-matched control cases revealed an overall decrease of 25% in total numbers of neurons in AD as compared with control eyes. Detailed analyses of GCL neurons at various eccentricities from the foveola showed that the greatest decrease in neuronal density (43% decrease) occurred in the central 0-0.5 mm (foveal region), while at 0.5-1 mm and at 1-1.5 mm eccentricities, neuronal loss amounted to 24 and 26%, respectively. The temporal region of the central retina appeared most severely affected, with up to 52% decrease in neuronal density near the foveola (central 0-0.5 mm eccentricity). There was close agreement between fellow eyes analyzed separately for three AD and three control cases. Analysis of neuronal sizes showed that all sizes of neurons were similarly affected in AD. In the GCL of control retinas, neurons decreased with age (coefficient of correlation = -0.67), while in AD retinas no such relationship was evident. Since in the central 0-2 mm region of the retina 97% of neurons in the GCL are ganglion cells (while the remaining 3% consist of displaced amacrine cells), these results demonstrate extensive ganglion cell loss in the central retina in AD. PMID- 8725900 TI - Retinal pathology in Alzheimer's disease. II. Regional neuron loss and glial changes in GCL. AB - Detailed analyses of neuronal and astrocyte cell numbers in the ganglion cell layer (GCL) of whole-mounted peripheral retinas from 16 Alzheimer's disease (AD) and 11 control eyes (11 and 9 cases, respectively) demonstrate extensive neuronal loss throughout the entire retina in AD as compared to control eyes. The observed neuronal loss is most pronounced in the superior and inferior quadrants, ranging between 40 and 49% throughout the midperipheral regions, and reaching 50-59% in the far peripheral inferior retina, while the overall neuronal loss throughout the entire retina amounts to 36.4% (p < 0.004). Although the 16% increase in astrocyte numbers is not significant, the ratio of astrocytes to neurons is significantly higher (82%; p < 0.0008) in AD as compared to normal retina (0.238 +/- 0.070 vs. 0.131 +/- 0.042). These results are strengthened by the close agreement (within +/- 15% of respective means) found between fellow eyes. Analysis of glial fibrillary acidic protein immunoreactivity (GFAP-ir) in sections of retinas from an additional 12 AD and 19 control cases show increased GFAP-ir with more extensive labeling of astrocytes in the GCL as well as increased labeling of Muller cell end-feet and radial processes in AD as compared to control retinas. The extensive loss of neurons documented in these retinas, accompanied by an increased astrocyte/neuron ratio, provides further support for the substantial involvement of the retina in AD. PMID- 8725901 TI - Alzheimer's disease and eyeblink conditioning: 750 ms trace vs. 400 ms delay paradigm. AB - Patients with probable Alzheimer's disease (AD) and hippocampal disruption are severely impaired in eyeblink classical conditioning (EBCC) in the 400 ms delay paradigm. Hippocampectomized rabbits are not impaired in the delay paradigm but perform poorly in the trace paradigm. It was anticipated that probable AD patients would be severely impaired in the 750 ms trace paradigm. In Study 1, probable AD patients were significantly impaired in the trace EBCC paradigm, but the sensitivity of the test was poorer than for the delay paradigm. In Study 2, probable AD patients tested in trace were tested in the delay paradigm 4 months later. Sensitivity for AD was also better for the delay paradigm. Rabbits and humans show behavioral parallels in the 400 ms delay paradigm but not in the 750 ms trace paradigm. The 400 ms delay EBCC paradigm was superior to the 750 ms trace paradigm for the detection of AD. PMID- 8725902 TI - Antibodies to amyloid beta protein (A beta) crossreact with glyceraldehyde-3 phosphate dehydrogenase (GAPDH). AB - In the present study, we characterized the epitope of a monoclonal antibody against purified amyloid plaque cores (Am-3). By immunocytochemical experiments, Am-3 stained cerebrovascular and senile plaque amyloid in brain sections of patients with Alzheimer's disease (AD) in a similar manner to that of antibodies against amyloid beta-protein (A beta). By Western blotting experiments, Am-3 recognized only a 35 kDa protein, which was revealed to be glyceraldehyde-3 phosphate dehydrogenase (GAPDH), and not A beta or beta amyloid precursor protein (beta PP). However, Am-3 recognized both GAPDH and purified native A beta in a dot-binding assay. Therefore, we concluded that Am-3 recognized both GAPDH and native A beta. Other monoclonal antibodies (6C6 and AmT-1) against the synthetic peptide corresponding to residues 1-28 of A beta also recognized these proteins. Because the amino acid sequences of these two proteins are not homologous, we propose that the crossreactivity between A beta and GAPDH is a consequence of their similar conformational epitopes. The possibility of crossreactions would complicate immunochemical and immunocytochemical studies of brain aging, AD and Down's syndrome. The implications of crossreactivity in developing immunological assays and in investigating the amyloid deposits of AD are discussed. PMID- 8725903 TI - Beta AP deposition and head trauma. AB - Head trauma is considered to be a risk factor for Alzheimer's disease, because a high prevalence of beta AP deposits has repeatedly been reported in patients who died within a few days following head injury. To evaluate this statement, we undertook two studies using immunohistochemistry for beta AP and found a surprisingly low prevalence of beta AP diffuse deposits. We first selected 23 patients aged 17-63 years, who died 0-76 days after head trauma. Using beta AP antibody at the usual dilution (1:100), we did not find any deposits. With a high concentration of antibody (dilution 1:2) we found beta AP diffuse deposits in one 46-year-old case. In a second study, 17 patients aged 60-79 years old, who died 1 35 days after head injury, were compared to a control group. We did not find any significant difference in the density of beta AP diffuse deposits between cases and controls using usual dilutions of beta AP antibody. The density of beta AP diffuse deposits was linked only to aging and the presence of senile plaques. PMID- 8725904 TI - Characterization of glial cultures from rapid autopsies of Alzheimer's and control patients. AB - We have developed isolated and mixed cultures of microglia, astrocytes, and oligodendrocytes from rapid (mean of 2 h 55 min) autopsies of nondemented elderly patients and patients with Alzheimer's disease, Parkinson's disease, and multiple sclerosis. Cultures were derived from both the corpus callosum (CC) and superior frontal gyrus (SFG). Cultured microglia phagocytosed latex beads, were reactive for Dil-acetylated low density lipoprotein, were immunoreactive for CD68 and major histocompatibility complex II markers, and were not immunoreactive for fibroblast, astrocyte, or oligodendrocyte markers. Cultured astrocytes included fibrous and protoplasmic types, were immunoreactive for GFAP, and were not immunoreactive for fibroblast, microglia, or oligodendrocyte markers. Cultured oligodendrocytes were poorly adherent, were slow to develop, were immunoreactive for galactocerebroside, and were not immunoreactive for fibroblast, microglia, or astrocyte markers. Because they are readily manipulated under controlled experimental conditions, and because they permit immediate access to individual cells and sets of cells from patients who have actually suffered the disease, these cultures may provide an important new tool for unravelling the etiology and pathogenesis of human CNS disorders. PMID- 8725905 TI - Effects of aging on the densities, numbers, and sizes of retinal ganglion cells in rhesus monkey. AB - We used sterological procedures that yield unbiased estimates to quantify the densities, numbers, and soma sizes of retinal ganglion cells in seven young adult and six old rhesus monkeys. The retinae were flat mounted so that we could determine whether there are different aging-related losses in different retinal regions. The mean (+/-standard deviation) total number of ganglion cells was 1,529,039 +/- 115,260 in young-adult retinae and 1,556,698 +/- 165,056 in old retinae, a difference that was not statistically significant. There also were no significant differences between young and old retinae in the densities or total numbers of ganglion cells in the four retinal quadrants, in four concentric retinal zones from fovea to peripheral retina, or in smaller hemiretinal regions of the concentric zones. Ganglion-cell soma sizes also did not differ significantly between young and old animals. Moreover, counts of the largest ganglion cells, which probably correspond to P alpha ganglion cells, revealed no selective loss of these cells with aging. These results are consistent with our previous anatomical and physiological studies of the LGN. Together they suggest that the retino-geniculate pathways are relatively unaffected by aging in the rhesus monkey. PMID- 8725906 TI - Age-related decrease in cholinergic synaptic transmission in three hippocampal subfields. AB - The present study was designed to examine the effect of age on cholinergic synaptic transmission in the three principal hippocampal subregions, and to assess whether these effects covary with age-related behavioral deficits. Young (3 week), adult (9 month), and old (24-27 month) rats were first tested on the Morris water task, and most of the old rats were severely impaired on the spatial version. The cholinergic slow epsp was induced by tetanic stimulation of stratum oriens or stratum granulosum, and recorded intracellularly in vitro from CA1 and CA3 pyramidal cells and granule cells in the fascia dentata (FD). The amplitude of the slow epsp was significantly reduced in old rats in all areas (CA1 59%; CA3 55%; and FD 56%). This age-related decrease was also present following the blockade of glutamatergic and GABAergic transmission, ruling out possible artifactual contributions from these systems to the change in the slow epsp. Our data suggest that functional cholinergic transmission is compromised in all areas of the hippocampus during normal aging. Few statistically significant correlations, however, were found between the age-related deficit in spatial learning and the decrease in cholinergic synaptic function. PMID- 8725907 TI - Effects of aging and chronic nimodipine on hippocampal binding of [3H]CGS 19755. AB - Previous studies have suggested that aging is associated with impaired behavioral performance and with decrements of N-methyl-D-aspartate (NMDA) receptors in the rat hippo-campus. Other studies have indicated that chronic treatment with nimodipine, a Ca2+ channel antagonist, prevents the age-related decline in performance by rats in behavioral tasks. Therefore, we tested whether nimodipine altered binding of [3H]CGS 19755 to hippocampal NMDA receptors in rats whose performance on a 14-unit T maze had been tested previously (14). No significant age difference was observed in [3H]CGS 19755 binding in hippocampi from old Fischer-344 rats (27 months) as compared with mature but not senescent rats (9 months); however, old rats that received chronic treatment with a low dose of nimodipine (20 mg pellets implanted subcutaneously twice during 70 days of treatment) showed higher levels of binding. A high dose of nimodipine (40 mg pellets implanted by the same route and at the same times as the low dose) was without effect on [3H]CGS 19755 binding, although aged rats given this treatment performed better in the maze than rats that received no nimodipine or the low dose. In a second experiment comparing hippocampi of young (4 months) and old (24 months) rats, saturation studies confirmed the lack of an age difference in [3H]CGS 19755 binding. The findings suggest that neither the age-related decline in maze performance nor the enhancement of behavior by nimodipine depend upon changes in hippocampal NMDA receptors. PMID- 8725908 TI - Age-related loss of calcium binding proteins in rabbit hippocampus. AB - Using immunocytochemistry hippocampal levels of the calcium binding proteins calbindin 28K (CB) and parvalbumin (PV) was studied in young (1 month) to very old (60 month) Albino rabbits. Young (3 month) and senescent (30 month) Wistar rats were also examined to compare the distribution and age dependency of PV and CB in both species. The distribution of PV-ir is similar in the rabbit and rat hippocampus. Aging in both species yielded a small loss of PV-ir in axon terminals. The presence of CB-ir interneurons throughout the hippocampus, and the heavy investment of the dentate gyrus (DG) granular cells with CB-ir was also similar in both species. In rabbits, the number of CB-ir interneurons in the CA1, as well as the density of CB-ir in the DG decreased in the first year of life, and did not change between 12-48 months of age. A secondary reduction in the density of CB-ir in the DG was observed at ages beyond 48 months. A similar loss of CB-ir in the DG occurred in the rat. In the CA1, however, the density of CB-ir was similar in young and aged rats. Another remarkable finding was the total absence of CB-ir in CA1 pyramidal neurons of rabbits at any age. Thus, the distribution and age dependency of PV-ir in the hippocampus is similar in both species. The decline of CB-ir in the DG with advancing age is very prominent and may be related to an altered calcium homeostasis in these cells. However, the absence of CB-ir in the CA1 of rabbits makes a causal role for CB in the functional decline of CA1 pyramidal cells during aging unlikely. PMID- 8725909 TI - Astrocytes, brain aging, and neurodegeneration. PMID- 8725910 TI - Role of estrogen and iron in neurodegeneration: open peer commentary to "Astrocytes, brain aging, and neurodegeneration" by H. Schipper. PMID- 8725911 TI - Astrocytes, brain aging, and neurodegeneration. PMID- 8725912 TI - Elusive roles for astrocytes in neurodegenerative disorders. PMID- 8725913 TI - Genotoxic elements in glial degeneration. PMID- 8725914 TI - Neurobiological substrates of behavioral decline: models and data analytic strategies for individual differences in aging. PMID- 8725915 TI - Individual differences in aging: implications for stereological studies of neuron loss. PMID- 8725916 TI - Brain-behavior linkages in aged rodent models: strategies for examining individual differences. PMID- 8725917 TI - Morphologic features of the normal human cadaveric spinal cord. AB - STUDY DESIGN: The cross-sectional area and diameter of the normal cadaveric spinal cord at each segmental level were measured, and the morphologic features were presented. OBJECTIVES: To provide accurate anatomic descriptions and morphometric data of the human spinal cord. SUMMARY OF BACKGROUND DATA: There is a large individual variation in human spinal cord size, and no authorized standard of measurement has been established. There have been few detailed descriptions of the normal morphologic features of the spinal cord. METHODS: The authors measured the cross-sectional area and diameter of the spinal cord at each segment from C2 and S3 in 12 cadaveric specimens, and the morphologic features of each segment were described. RESULTS: The relative ratio of the cross-sectional area of each segment to that of the C3 segment was similar in all the specimens examined despite a large individual variation in absolute cord size. Each segment had distinct qualitative and quantitative morphologic features. CONCLUSIONS: The normal cross-sectional area of the spinal cord at any segment in an individual is calculable from measurements of a given single normal segment. This value appears to be an appropriate and practical standard of measurement of the normal morphologic features of the spinal cord. PMID- 8725918 TI - The location of the vertebral artery foramen and its relation to posterior lateral mass screw fixation. AB - STUDY DESIGN: This study evaluated the anatomic relationship between the vertebral artery foramen and the posterior midpoint of the cervical lateral mass using cervical spine specimens. OBJECTIVES: To determine quantitatively the location of the vertebral artery foramens from C3 to C6 and their relationship to the posterior midpoints of the lateral masses. SUMMARY OF BACKGROUND DATA: Anatomic studies of the cervical nerve root and facet relative to lateral mass screw placement have been addressed. It is necessary to know the correct location of the vertebral artery foramen during lateral mass screw placement to minimize the risk of injury to the vertebral artery. METHODS: Forty-three cervical spines from C3 to C6 were directly evaluated for this study. Anatomic evaluation included the dimension of the vertebral artery foramen and its projection on the posterior aspect of the lateral mass. The vertical distance from the posterior midpoint of the lateral mass to the posterior border of the vertebral artery foramen, and the angle between the parasagittal plane and the line connecting the posterior midpoint of the lateral mass with the lateral limit of the vertebral artery foramen, were also measured. RESULTS: The vertical distances from the posterior midpoint of the lateral mass to the vertebral artery foramens at C3-C6 averaged from 9.3 to 12.2 mm for male and female specimens. The average angles medial to the sagittal plane, between the parasagittal plane and the line connecting the posterior midpoint of the lateral mass with the lateral limit of the vertebral artery foramen, from C3 to C5, were found to range from 6.0 degrees to 6.3 degrees for male specimens and from 5.3 degrees to 5.5 degrees for female specimens. At C6, the average angles lateral to the sagittal plane, between the parasagittal plane and the line connecting the posterior midpoint of the lateral mass with the lateral limit of the vertebral artery foramen, were 6.4 degrees for male specimens and 5.4 degrees for female specimens. CONCLUSIONS: The present study indicated that there is no risk of damaging the vertebral artery if a screw is directed perpendicular to the posterior aspect of the lateral mass at C3-C5 and 10 degrees lateral to the sagittal plane at C6 starting at the midpoint of the lateral mass. PMID- 8725919 TI - Projection of the lumbar pedicle and its morphometric analysis. AB - STUDY DESIGN: This study defined the projection point of the lumbar pedicle on its posterior aspect and its relation to a reliable landmark and reported pedicle dimensions based on 50 lumbar spines. OBJECTIVES: To establish the best starting point for a pedicle screw for passing the screw down the center (axis) of the pedicle; to describe quantitatively the relations of the pedicle projection point to a reliable landmark; and to evaluate the linear and angular dimensions of the lumbar pedicle. SUMMARY OF BACKGROUND DATA: Posterior transpedicular screw fixation has been most widely used for management of the unstable lumbar spine. Several studies of pedicular anatomy exist, but little quantitative data regarding the location of the lumbar pedicle axis for each level have been reported. METHODS: Fifty dry lumbar specimens (250 lumbar vertebrae) were obtained for study of the lumbar pedicle. Anatomic evaluation focused on determination of the projection point of the lumbar pedicle axis on the junction of the superior facet and the transverse process and measured the distance from the projection point to the midline of the transverse process for each level of the lumbar vertebrae. Pedicle dimensions, including linear and angular, also were measured. RESULTS: Differences in dimensions between men and women were not found to be statistically significant. The average distance from the projection point to the midline of the transverse process consistently changed from L1 to L5. Above L4, the projection point for men and women averaged 3.9 mm for L1, 2.8 mm for L2, and 1.4 mm for L3 superior to the midline of the transverse process, respectively. At L4, the projection point was close to the midline of the transverse process (0.5 mm inferior). At L5, the projection point was an average of 1.5 mm inferior to the midline of the transverse process. CONCLUSIONS: The average distance from the projection point of the lumbar pedicle axis to the midline of the transverse process consistently varied at different levels. This information may prove helpful in the placement of screws into the lumbar pedicle. PMID- 8725921 TI - The effect of fibrin sealant on spinal fusions using allograft in dogs. AB - STUDY DESIGN: This study investigated the use of Tisseel (immuno [Canada], Toronto, Ontario) as an adjunct to allograft spinal fusion. Thirteen mongrel, dogs were fused bilaterally with morcellized graft from a separate dog. OBJECTIVES: To evaluate whether fibrin sealant had an effect on bone volume of fusion mass in allograft fusions of the spine. SUMMARY OF BACKGROUND DATA: Fibrin sealant has been promoted for use in many orthopedic applications. There is controversy about its effectiveness in augmenting bone graft healing. However, some surgeons make routine use of the sealant in augmentation of bone grafting procedures. METHODS: To test the usefulness of this material in augmenting allograft fusions, the authors carried out bilateral posterolateral fusions in 13 mongrel dogs. At surgery, 15 cm3 of allograft was placed into a posterolateral position at the L5-L6 region on both sides of the spine. Fibrin sealant (Tisseel) was allocated randomly to one side only. Fusion mass was tested 6 months after the initial operation by computed tomographic scan imaging and mechanical testing. RESULTS: A significantly smaller bone volume mass, as illustrated by computed tomographic measurement, was seen on the Tisseel side (P = 0.03). Biomechanical testing indicated that there was a trend for the Tisseel side to be stiffer than the untreated side, particularly at lower weights, but statistical significance was not achieved. Computed tomographic volumetric analysis showed that Tisseel-treated allograft led to a significantly smaller fusion volume. CONCLUSIONS: This study refutes the belief that Tisseel is a good material for accomplishing or augmenting intervertebral arthrodesis. Fibrin sealant significantly retards allograft fusion mass formation in dogs. PMID- 8725920 TI - Immunohistochemical demonstration of sensory and autonomic nerve terminals in herniated lumbar disc tissue. AB - STUDY DESIGN: Thirty-five lumbar disc herniations removed at surgery were studied by indirect immunocytochemistry. OBJECTIVES: To localize immunohistochemically both sensory and autonomic nerve terminals in disc herniations. SUMMARY OF BACKGROUND DATA: Using various more or less specific histologic and histochemical methods, investigators have reported the presence of free nerve terminals in disc tissue. However, very few studies have, to date, convincingly demonstrated nerve terminals in disc tissue that morphologically resemble the tiny nerve terminals of sensory and autonomic nerve fibers. METHODS: Amplification of the peroxidase reaction product in avidin-biotin-peroxidase complex immunostaining by the glucose oxidase-diaminobenzidine-nickel sulfate method was used to visualize small punctate nerve terminals at high magnification. Thin frozen sections from disc herniation tissue prefixed in Zamboni fixative were incubated with antibodies to synaptophysin to visualize nerve terminals in general, and with antibodies to substance P and C-flanking peptide of neuropeptide Y to further characterize nerve terminals as either sensory or sympathetic. RESULTS: Nerve terminals could be demonstrated in 29 (83%) of the 35 disc herniations. They were observed with the synaptophysin antibody in 17 of 35 (49%) disc herniations, with substance P in 16 of 35 (46%) disc herniations, and with C-flanking peptide of neuropeptide Y in 13 of 35 (37%) disc herniations. Morphologically, the nerve terminals were seen as tiny immunoreactive dots. Some of the nerve terminals were observed close to disc cells, possibly suggesting direct interaction. CONCLUSIONS: Small nerve terminals in disc herniations, both sensory substance P endings and sympathetic C-flanking peptide of neuropeptide Y endings, could be involved in mechanisms of discogenic pain, disc tissue neurogenic inflammation, tissue repair processes after injury, and control of local blood circulation in the newly formed blood vessels. Disc cells may be directly affected by the neuropeptides released from nearby nerve terminals. PMID- 8725922 TI - Changes in cervical canal spinal volume during in vitro flexion-extension. AB - STUDY DESIGN: Quasistatic flexion and extension loads were applied in vitro to lower cervical spines. The flexion-extension motion produced was checked for physiologic relevance. OBJECTIVES: To examine the changes in the volume of the cervical spinal canal in flexion-extension motion. SUMMARY OF BACKGROUND DATA: Many papers have been published concerning the cervical canal volume as inferred from standard lateral radiographs. This study compares the inferred (radiographic) volumes and their changes to the physical changes within the spinal canal. METHODS: The lower cervical spines (C2-C7) from 10 cadavers were subject to stepwise flexion and extension in a purpose-built rig. Before this testing, the spinal cord was removed from the canal space of each specimen and replaced by a thin latex tube stoppered and secured at the opening of the canal (at C2) so that the volume of liquid displaced from the tube could be measured. This was done at each loading stage by means of a graduated glass column, and a radiograph of the spine was also taken to allow angular and displacement readings to be taken from C2 to C7. RESULTS: The average recorded change in volume of the spinal canal with flexion-extension motion was 1.9 ml, and showed a significant linear correlation with the dynamic canal width (r = 0.868, P < 0.05) and also with the total angle of flexion or extension (r = 0.979, P < 0.005). The volume of liquid displaced from the canal in lateral bending was much lower than that in flexion-extension motion, and only amounted to about 0.2 ml. The angular ranges of motion produced at each level were compared to previous results obtained in vivo, and no significant differences between the angular displacements found in vivo and in vitro under this experimental arrangement were seen. CONCLUSIONS: The loading regime described in this study causes angular displacements similar to those in vivo, and on this basis is a physiologically relevant loading pattern. The change in the volume of the spinal canal between C2 and C7 shows linear relationships with the angle of flexion and the dynamic canal width. PMID- 8725923 TI - Accuracy of pedicle screw placement in lumbar vertebrae. AB - STUDY DESIGN: The location of pedicle screws (n = 42) in four human specimens of the lumbar spine and in 30 patients (n = 131 screws) after lumbar spinal fusion was assessed using computed tomography. OBJECTIVES: To determine the accuracy of pedicle screw placement in lumbar vertebrae and the reproducibility and repeatability of the computed tomography examination. SUMMARY OF BACKGROUND DATA: Failures in the placement of transpedicular screws for lumbar fusion are reported. The evaluation of such screws using computed tomography examination has not been investigated. METHODS: After surgery, the specimens were dissected in transversal slices to observe macroscopically the location of the pedicle screw and to correlate these observations with the computed tomography images. All patients were examined by one observer. To determine the reproducibility and repeatability of the computed tomography examination, two observers studied computed tomography images of 12 patients (n = 58 screws) twice within 3 months. RESULTS: In the specimens, 10 screws were observed to penetrate the medial wall of the pedicle. This correlated fully with the images. In the patients' group, 40% of all screws penetrated the cortex of the vertebra. Of all screws, 29% penetrated the medial wall of the pedicle. From the computed tomography images, it appeared that a deviation of more than 6 mm medially was a high risk for nerve root damage. Three months after his first examination, Observer 1 documented a different position in three of 58 screws (kappa = 0.90). Observer 2 found a different position in eight screws (kappa = 0.65). The comparison between the reviews of the two observers showed a different opinion for the first evaluation, four disagreements (2-4 mm) and 17 disagreements (0-2 mm; kappa = 0.34), and for the second evaluation, four disagreements (2-4 mm) and 12 disagreements (0-2 mm; kappa = 0.43). CONCLUSIONS: Correct placement of transpedicular screws for spinal fusion seems to be more difficult than it looks. The computed tomography scanning is useful for differential diagnosis of postoperative radicular syndromes after lumbar transpedicular fixation. PMID- 8725924 TI - Diagnostic accuracy of technologies used in low back pain assessment. Thermography, triaxial dynamometry, spinoscopy, and clinical examination. AB - STUDY DESIGN: A prospective blind study compared three new technologies to assess back pain. OBJECTIVE: To assess the diagnostic accuracy and comparability of thermography, triaxial dynamometry, and spinoscopy in the assessment of recent onset work-related low back pain. SUMMARY OF BACKGROUND DATA: The role of these technologies in assessing patients with low back pain is unproved. METHODS: Forty one patients with low back pain and 46 control subjects were assessed by each technology and by two clinical examiners blind to clinical status. Twenty patients were trained to simulate a healthy back without low back pain, and 50% of the control subjects were trained to simulate the presence of a low back pain disorder. Each technology was interpreted on two occasions by each of two readers. RESULTS: Thermography performed significantly worse than did triaxial dynamometry, spinoscopy, and clinical examination. The diagnostic accuracy of the last three was similar, and inter-rater comparability did not differ significantly. Among simulators, the diagnostic accuracy of triaxial dynamometry and spinoscopy was significantly higher than that of clinical examination, although considerable inaccuracy remained in assessing individual subjects. CONCLUSIONS: The diagnostic accuracy of thermography in recent onset low back pain does not support its use. Among those simulating normality or low back pain, triaxial dynamometry and spinoscopy have greater diagnostic accuracy than does a single clinical evaluation. However, for an individual, the inaccuracy that remains limits the use of triaxial dynamometry or spinoscopy for diagnosis in recent onset low back pain. PMID- 8725925 TI - Lumbar range of motion: reliability and validity of the inclinometer technique in the clinical measurement of trunk flexibility. AB - STUDY DESIGN: This study examines the reliability and validity of measuring lumbar range of motion with an inclinometer. OBJECTIVES: To find out whether a manual determination of the reference points for measuring lumbar range of motion is as reliable as radiologic determination for positioning the inclinometers, lumbar range of motion was determined in degrees by evaluating radiographs and by using the inclinometer technique of Loebl. SUMMARY OF BACKGROUND DATA: Reliability and validity of the inclinometer technique as a clinical measurement of trunk flexibility were investigated. Fifty-four patients participated in the study. METHODS: Lumbar range of motion measurements were taken with and without radiologic control of the T12 and S1 vertebrae as reference points for positioning of the inclinometers. An interrater correlation was done of the inclinometer techniques of a physician and a physiotherapist. Functional radiographs were investigated in a standing position. Lumbar range of motion measurements based on radiographs and those taken using the inclinometer alone were correlated to validate the inclinometer technique. RESULTS: Lumbar range of motion measurements taken with and without radiologic determination showed a very close correlation (r = 0.93; P < 0.001). Flexion alone also demonstrated a close correlation (r = 0.95; P < 0.001), whereas extension showed a somewhat smaller correlation (r = 0.82; P < 0.001). Total lumbar range of motion (r = 0.94; P < 0.001) and flexion (r = 0.88; P < 0.001) were closely related, as indicated by the interrater correlation, whereas extension (r = 0.42; P < 0.05) showed a lesser correlation. Correlation of the measurements taken radiographically and by inclinometer demonstrated an almost linear correlation for measurements of the total lumbar range of motion (r = 0.97; P < 0.001) and flexion (r = 0.98; P < 0.001), whereas extension (r = 0.75; P < 0.001) did not correlate as well. CONCLUSIONS: The noninvasive inclinometer technique proved to be highly reliable and valid, but the measurement technique for extension needs further refinement. PMID- 8725926 TI - Lumbar range of motion: influence of time of day and individual factors on measurements. AB - STUDY DESIGN: Influence of time of day and individual factors on the measurements of the lumbar range of motion was investigated. OBJECTIVES: To investigate factors that influence lumbar range of motion. SUMMARY OF BACKGROUND DATA: Twenty nine patients with chronic low back pain participated in the study. METHODS: The lumbar range of motion was measured by inclinometer technique, Schober sign, modified-Schober sign, and fingertip-to-floor method in the morning, at noon, and in the afternoon. The lumbar range of motion was correlated with patients' gender, age, and body weight. RESULTS: Total lumbar range of motion measured by the inclinometer technique and the modified-Schober sign increased significantly throughout the day from morning to afternoon. Extension lumbar range of motion was shown to be independent of the time of measurement. There were no significant correlations between lumbar range of motion and gender, age, and body weight of the patients. CONCLUSION: For the reliability of a measurement, it is important to investigate lumbar range of motion at the same time of day. PMID- 8725927 TI - Prospective outcome evaluation of spinal cord stimulation in patients with intractable leg pain. AB - STUDY DESIGN: The results of spinal cord stimulation were prospectively evaluated using both subjective patient self-report measures and objective physical functional testing. OBJECTIVES: The purpose of this study was to evaluate prospectively the effects of spinal cord stimulation implantation, performed with the patient awake and providing feedback, in patients with primary reports of intractable leg pain. SUMMARY OF BACKGROUND DATA: Spinal cord stimulation has been used for treating chronic pain of many types. However, even among those with intractable lower extremity pain, the outcome results have shown great variability. METHODS: The surgical procedure was performed with the patient awake and providing feedback to ensure optimal pain relief from the lead placement. The study group comprised 40 patients, ranging in age from 28 to 86 years. The average symptom duration was 65.4 months, and the average number of prior lumbar spine surgeries was 2.3 (range, 1 to 8). The primary data collection periods were preoperative, 6 weeks after, and 12 and 24 months after surgery. RESULTS: Statistically significant improvement in isometric lower extremity function was demonstrated 6 weeks after the spinal cord stimulation implantation. In the more painful leg, the performance increased from 457.5 ft-lb-sec to 629.8 ft-lb-sec (P < 0.01). The performance remained significantly improved at the 12- and 24-month follow-ups. Significant improvement was demonstrated on the physical scale of the Sickness Impact Profile at 6 weeks. At 24 months, all three scales (physical, psychological, and other) as well as the total score were significantly improved. Statistically significant decreases in pain, assessed by changes in visual analog scale scores, were noted in the legs, when walking, and in overall lifestyle. The use of narcotic medication decreased at all follow-up periods. At least 66% of the patients who were taking narcotics before spinal cord stimulation were taking reduced amounts or no narcotics 2 years later. At the time of the 24-month follow up, at least 70% of patients reported that the procedure helped them, and would recommend it to someone with similar symptoms. CONCLUSIONS: Spinal cord stimulation implantation can result in improved physical function and decreased pain in patients who are carefully screened and in whom the implantation is performed with the patient awake to help ensure optimal pain-relieving lead placement. PMID- 8725928 TI - Long-term results of partial undercutting facetectomy for lumbar lateral recess stenosis. AB - STUDY DESIGN: A retrospective review of 57 consecutive patients who had a partial undercutting facetectomy for degenerative lumbar lateral recess stenosis between 1983 and 1988. OBJECTIVES: To evaluate the long-term results of this procedure. SUMMARY OF BACKGROUND DATA: Few studies have been published on the long-term outcome of decompression for lumbar stenosis. Most studies have included central as well as lateral recess stenosis and have not differentiated between the two. Furthermore, no study has looked at the long-term results of partial undercutting facetectomy. METHODS: All patients were assessed by standard questionnaire, clinical examination, and radiography by an independent observer. The minimum follow-up period was 5 years (mean, 8.4 years). RESULTS: Overall, 72% had no leg pain, 16% had some leg pain needing occasional analgesia, and 12% had severe leg pain needing continual analgesia. CONCLUSIONS: The long-term results of partial undercutting facetectomy are very satisfying. PMID- 8725929 TI - Pelvic fractures after long lumbosacral spine fusions. AB - STUDY DESIGN: A retrospective review of late pelvic ring fractures after long spine fusions to the lumbosacral spine. OBJECTIVES: To review the clinical course and predisposing features of late fractures of the pelvic ring, usually atraumatic, in patients with long fusions to the lumbosacral spine. SUMMARY OF BACKGROUND DATA: Fractures of the pelvic ring after long fusions to the lumbosacral spine is a heretofore rarely reported complication. METHODS: Records from the authors' institution from 1985 to 1994 were reviewed retrospectively. Two hundred sixty-eight patients with long fusions to the lumbosacrum were identified. RESULTS: Between 1985-1994, five patients suffered late atraumatic fractures of the pelvic ring after long instrumented fusions to L5 or the sacrum. All fractures were on the left side of the pelvic ring, primarily the public rami. All patients were women, and at the time of fracture, all were aged 50 years or older. Fractures occurred from 4 months to 7 years after the last surgery (mean, 28 months). All were treated with protected weightbearing until comfortable. At average 27 months' follow-up evaluation, four patients reported no pain; the fifth had become pain-free at 31 months, but 2 months later, the patient spontaneously fractured the contralateral public rami. CONCLUSION: Stress type fractures of the pelvic ring are a potential source of late pain after long fusions to the lumbosacral spine. Orthopedic surgeons should be aware of this possibility, especially in older, potentially osteoporotic women. Early treatment with simple protected weightbearing appears satisfactory and can prevent significant morbidity. PMID- 8725930 TI - Understanding peripartum pelvic pain. Implications of a patient survey. AB - STUDY DESIGN: An analysis was made of the self-reported medical histories of patients with peripartum pelvic pain. OBJECTIVES: To compile an inventory of the disabilities of patients with peripartum pelvic pain, analyze factors associated with the risk for development of the disease, and to formulate a hypothesis on pathogenesis and specific preventive and therapeutic measures. SUMMARY OF BACKGROUND DATA: Pregnancy is an important risk factor for development of chronic low back pain. Understanding the pathogenesis of pelvic and low back pain during pregnancy and delivery could be useful in understanding and managing nonspecific low back pain. METHODS: By means of a questionnaire, background data were collected among patients of the Dutch Association for Patients With Pelvic Complaints in Relation to Symphysiolysis. Results were compared with the general population. Subgroups were compared with each other. RESULTS: Peripartum pelvic pain seriously interferes with many activities of daily living such us standing, walking, sitting, and all other activities in which the pelvis is involved. Most patients experience a relapse around menstruation and during a subsequent pregnancy. Occurrence of peripartum pelvic pain was associated with twin pregnancy, first pregnancy, higher age at first pregnancy, larger weight of the baby, forceps or vacuum extraction, fundus expression, and a flexed position of the woman during childbirth; a negative association was observed with cesarean section. CONCLUSIONS: It is hypothesized that peripartum pelvic pain is caused by strain of ligaments in the pelvis and lower spine resulting from a combination of damage to ligaments (recently or in the past), hormonal effects, muscle weakness, and the weight of the fetus. PMID- 8725931 TI - Superior gluteal artery injury secondary to posterior iliac crest bone graft harvesting. A surgical technique to control hemorrhage. AB - STUDY DESIGN: This case series and cadaveric dissection illustrates a method of obtaining hemostasis of iatrogenic superior gluteal vessel injury sustained during posterior iliac crest bone graft harvesting. OBJECTIVES: To show a simple and effective method of obtaining hemostasis of the iatrogenic superior gluteal vessel injury associated with posterior iliac crest bone graft harvesting. SUMMARY OF BACKGROUND DATA: Management of superior gluteal vessel injury has included direct pressure, enlargement of the sciatic notch to allow for exposure of bleeding vessels, retroperitoneal or transperitoneal approaches, and angiographic embolization to obtain hemostasis. The authors present several cases and a cadaveric study to show a simple and effective technique used to control hemorrhage secondary to iatrogenic superior gluteal vessel injury sustained at the time of posterior iliac crest bone graft harvesting. METHODS: The management of iatrogenic superior gluteal vessel injury secondary to posterior iliac crest bone graft harvesting involved the extension of the surgical incision, detachment of the origin of the gluteus maximus, lateral retraction of the gluteus maximus along with the tethered superior gluteal vessels, and visualization and ligation of the injured vessels. RESULTS: Hemostasis was achieved quickly with minimal loss of blood. Additional surgery or angiographic embolization was not required. CONCLUSIONS: In the cases presented, extension of the posterior iliac bone graft incision, detachment of the origin, and reflection of the gluteus maximus provided excellent exposure and hemostasis of the iatrogenic laceration of the superior gluteal artery. This technique is simple and effective and may prevent the need for transperitoneal and retroperitoneal approaches or angiographic embolization. PMID- 8725932 TI - Bilateral vertebral artery lesion after dislocating cervical spine trauma. A case report. AB - STUDY DESIGN: This case report illustrates the problems associated with diagnosis and management of vertebral artery injuries resulting from dislocating cervical spine trauma. OBJECTIVES: Treatment involved the principles of anterior stabilization of dislocating cervical spine fracture as well as the diagnostic procedures and therapeutic modalities appropriate for vertebral artery lesions. SUMMARY OF BACKGROUND DATA: Because vertebral artery injuries with cervical spine trauma are rarely symptomatic, they can easily be overlooked. Bilateral or dominant vertebral artery occlusion, however, may cause fatal ischemic damage to the brain stem and cerebellum. METHODS: Cervical spine dislocation was stabilized immediately after admission using internal fixation by ventral plate and corticocancellous bone graft. Immediate angiography was performed when brain stem neurologic dysfunction manifested 36 hours after surgery. The patient was treated with anticoagulation, osmotherapy, and controlled hypertension. RESULTS: A fatal outcome resulted in this case of dominant left vertebral artery occlusion. Necropsy even revealed bilateral vertebral artery damage at the level of the osseous lesion. CONCLUSIONS: The possibility of the complication of a vertebral artery lesion should be kept in mind when examining patients with cervical spine trauma, especially in patients with fracture-dislocation. Immediate identification by vertebral angiography, magnetic resonance imaging, or thin slice computed tomography scan is necessary for optimal management of this injury. PMID- 8725933 TI - Hematocolpos. An unusual cause of sciatica in an adolescent girl. AB - STUDY DESIGN: A case report. OBJECTIVE: To document the occurrence of hematocolpos causing sciatica and leg weakness in an adolescent girl. SUMMARY OF BACKGROUND DATA: Low back pain resulting from hematocolpos has been reported previously but not associated with symptoms of sciatica or neurologic weakness. METHODS: The authors were involved in the care of this patient, and all medical records, radiologic investigations, and related literature were reviewed. RESULTS: After being checked for longstanding symptoms of sciatica and leg weakness, the patient underwent a computed tomographic examination of lumbar spine, which showed a large collection in the pelvis consistent with hematocolpos. Surgical drainage of the collection was associated with complete resolution of her symptoms. CONCLUSIONS: Awareness of hematocolpos as a possible cause for sciatica and neurologic symptoms in the lower limbs may lead to earlier diagnosis and, therefore, reduced morbidity in the investigation of the adolescent girl. PMID- 8725934 TI - Results of surgery compared with conservative management for lumbar disc herniations. AB - Conservative management gives satisfactory results in a high proportion of patients with disc herniation in the course of a few months of treatment onset. This is likely to occur particularly in patients with mild or moderate nerve root compression. Surgical treatment is significantly faster in yielding a satisfactory resolution of symptoms. The chances of a successful outcome are higher in patients with a marked nerve root compression, no or mild back pain, and a short duration of symptoms. The results of surgery often deteriorate in the long and very long term because of recurrence of radicular, and especially low back, pain. In most patients, deterioration is not related strictly to surgery because chances of pain recurrence or worsening with time are similar in the patients treated conservatively and in those who undergo surgery. Microdiscectomy appears to give slightly better results than standard operation in the first few weeks or months after surgery, but not successively. Surgery usually should be undertaken if the patient does not respond after at least 2 months' conservative treatment. PMID- 8725935 TI - Andreas Vesalius 1514-1564. AB - Andreas Vesalius was born in Brussels on December 31, 1514. After having spent some disappointing years at the Universities of Louvain and Paris, he graduated as Doctor of Medicine in Padua on December 5, 1537. The next day he was appointed as a teacher of both human anatomy and surgery. During the 6 years he held this chair, Vesalius engaged in impressive academic activities and published three masterly anatomic books: Tabulae Anatomicae Sex, De Humani Corporis Fabrica Libri Septem, and Epitome. The last two works contain anatomic woodcuts of incomparable artistic quality by Titian's pupils (by Stefan v. Calcar in particular). In 1544, at the age of 28, Vesalius gave up his chair and took up service as a court physician, first with Emperor Charles V and later with his son, Philip II of Spain. He died in 1564 on the small Greek island of Zante on return from a pilgrimage to the Holy Land. The gist of Vesalius' teaching was his conviction that valid anatomic knowledge could be gained only through dissection of the human corpse and not through the study of the traditional texts. Vesalius rid the study of human anatomy of mythic speculations, which had encrusted it for two millennia. Through Vesalius' work, human anatomy became an empirical science. Like Copernicus, Kepler, Bruno, and Galileo, Vesalius was one of the initiators of the new science. The tables of osteology and of the spine in Fabrica and Epitome are most impressive. Much of the nomenclature used for the spine today can be credited to him. PMID- 8725936 TI - Outcome of unoperated discogram positive low back pain. PMID- 8725937 TI - Lymphocyte development. PMID- 8725938 TI - Gene targeting reveals a hierarchy of transcription factors regulating specification of lymphoid cell fates. AB - Specification of B- and T-lymphoid cell fates appears to involve the expression of a shared set of genes encoding recombination proteins and of genes encoding lineage-specific components of antigen receptors. Recent studies using gene targeting have identified transcription factors that are required for the proper specification of lymphoid cell fates. On the basis of these data, a regulatory gene hierarchy which orchestrates the development of lymphoid progenitors from hematopoietic stem cells can be proposed. PMID- 8725939 TI - Regulation of gene expression at early stages of B-cell and T-cell differentiation. AB - The expression of distinct sets of genes at different stages of B-lymphocyte and T-lymphocyte differentiation is controlled at the level of transcription. A number of recent studies have described interactions between transcription factors in lymphocytes that provide new insights into mechanisms regulating gene expression. These mechanisms include the assembly of higher order nucleoprotein complexes and other protein-protein interactions that enhance the functional specificity of transcriptional regulators in lymphocytes. PMID- 8725940 TI - Mechanism of V(D)J recombination. AB - V(D)J recombination can be separated into two basic operations: DNA cleavage and joining of broken ends. Our understanding of both reactions has increased substantially in the past year. Major advances include the development of a cell free system capable of cleavage and the identification of several proteins involved in both V(D)J recombination and double-strand break repair. PMID- 8725941 TI - Composition and function of T-cell receptor and B-cell receptor complexes on precursor lymphocytes. AB - The precursor T-cell receptors (TCRs) and B-cell receptors (BCRs) direct lymphocyte development to the mature T-cell and B-cell stage, respectively. Recent genetic and biochemical experiments reveal the striking parallel in structure and function of these receptors. They consist of TCR beta and BCR mu chains paired with surrogate TCR alpha and BCR light chains. Both receptors employ a two-component signal transduction unit: CD3 gamma epsilon for the pre TCR, and CD79ab for the pre-BCR. Plasma membrane levels of pre-TCR/BCR complexes are kept extremely low, most probably by a mechanism involving specific retention in the endoplasmic reticulum. This mechanism may control the signalling activity of pre-TCR/BCR and therewith the lymphocyte differentiation process. PMID- 8725942 TI - Signalling in lymphocyte development. AB - A number of important signal-transduction molecules that regulate lymphocyte maturation and proliferation have been identified. These advances provide a platform for studies on how different signalling events are integrated to generate the required number of lymphocytes with an appropriate antigen receptor repertoire. PMID- 8725943 TI - Immunoglobulin class switching. AB - Antibody class switching is induced by B-cell activators in the presence of cytokines. The identity of the heavy-chain class to which a B cell is switched is regulated by cytokines and B-cell activators at the level of transcription of unrearranged heavy chain constant genes. Gene-targeting experiments in mice have proved the essential role of these transcripts in switch recombination. Their possible functions are discussed in the context of a model for the mechanisms of class switching. PMID- 8725944 TI - The molecular basis of somatic hypermutation of immunoglobulin genes. AB - Somatic hypermutation amplifies the variable region repertoire of immunoglobulin genes. Recent experimental evidence has thrown light on various molecular models of somatic hypermutation. A link between somatic hypermutation and transcription coupled DNA repair is shaping up. PMID- 8725945 TI - Regulation of thymocyte development from immature progenitors. AB - T lymphocytes differentiate from hematopoietic stem cells that settle in the microenvironment of the thymus. The earliest stages of mouse alpha/beta T-cell differentiation occurring before surface expression of the TCR include three important events: proliferation, commitment to the T lineage, and rearrangement and expression of the TCR loci. Recent evidence suggests that the survival as well as differentiation of early thymocytes depends critically on molecular signals such as those generated by the recently described pre-TCR complex. PMID- 8725946 TI - Positive selection of CD4+ and CD8+ T cells. AB - Significant progress has been made in characterizing intermediates and defining individual steps of positive selection, providing important insights into mechanisms of CD4/CD8 lineage commitment. New evidence suggests that specific recognition of peptides may be important for positive selection of CD4+ T cells. Several studies have defined signal-transduction pathways important for positive selection and have provided evidence that distinct signaling pathways may regulate positive versus negative selection. PMID- 8725947 TI - Mechanisms underlying T-cell tolerance. AB - To understand how the immune system manages to disarm potentially harmful T cells with reactivity against self-antigens, many immunologists have turned to model systems in which transgenic expression of TCRs, foreign antigens, or both, allows visualization of otherwise undetectable mechanisms. Recent data have offered several new insights into the variables that dictate the mechanism for tolerance employed by peripheral T cells. Some of these mechanisms can also dampen ongoing immune responses against foreign antigens and provide a strikingly powerful supplement to the primary mechanism of tolerance, thymic clonal deletion. PMID- 8725948 TI - Apoptosis and the maintenance of homoeostasis in the immune system. AB - The regulation of cell proliferation and the selection against autoreactive cells in the lymphoid system both occur through the induction of apoptosis. Many of the signals that induce apoptosis in lymphocytes are now well defined. Interactions between Fas and its ligand have emerged as a major mechanism for the deletion of activated peripheral T cells and autoreactive B cells. Although the signal transduction pathway leading from engagement of Fas to apoptosis is not entirely clear, significant advances have been made recently. There has also been progress in the elucidation of the mechanisms that regulate apoptosis in the immune system. PMID- 8725949 TI - Analysis of cell-adhesion molecule interactions using surface plasmon resonance. AB - The molecular interactions that mediate cell adhesion are often very weak, making them difficult to study. However, real-time optical biosensors based on surface plasmon resonance (SPR) are greatly facilitating the biochemical analysis of these interactions. Analysis of the T cell surface molecule CD2 has shown that adhesion molecules can interact with very low affinities (Kd approximately 100 microM) and dissociate with half lives of approximately 0.2 seconds or less. SPR has been combined with site-directed mutagenesis to delineate the interacting surfaces of CD2 and its ligand, CD48, quantify the contribution of individual residues to the binding energy, and determine the binding orientation of these surfaces in the CD2-CD48 complex. Furthermore, SPR has been combined with in situ modification of carbohydrates on purified glycoproteins to analyze the binding specificity of lectins such as CD22. Researchers have discovered the potential pitfalls of SPR, which can lead to inaccurate affinity and kinetic measurements. PMID- 8725950 TI - Studying interactions involving the T-cell antigen receptor by surface plasmon resonance. AB - T-lymphocyte activation is initiated by the interaction of the alpha beta TCR with a complex consisting of a class I or class II MHC-encoded molecule and an antigenic peptide, displayed on the surface of an antigen-presenting cell. Real time binding measurements using surface plasmon resonance have revealed kinetic and equilibrium parameters for the interactions between purified MHC molecules and peptides, between TCR and MHC-peptide complexes, and between TRC and superantigens. The MHC-peptide interaction is characterized by its high affinity and long half-life, the TCR-MHC/peptide interaction by its low affinity and short half-life, and the TCR-superantigen interaction by its low-to-moderate affinity, which is dependent on the particular superantigen involved. The consistent finding is that both MHC-peptide complexes and superantigens interact with TCR with a low affinity attributable to rapid dissociation. That an MHC-peptide complex that encounters a single TCR only briefly can still deliver the necessary activation signals offers a mechanistic conundrum for which several solutions have been proposed. PMID- 8725951 TI - Needles in haystacks: identifying specific peptide antigens for T cells. AB - The antigen receptors of T cells are engaged by unique peptides displayed by MHC molecules on the antigen-presenting cell surface. These rare peptides can now be distinguished among the thousands normally bound to MHC molecules. Knowledge of the antigenic peptides and their donor proteins is suggesting potential targets for immunotherapy and providing insights into the cellular mechanisms that generate the peptide-MHC complexes. PMID- 8725952 TI - Tracking antigen-specific helper T cell responses. AB - T-cell receptor transgenic animals provide an excellent source of T cells for the analysis of antigen-specific helper T-cell development. Alternatively, studies in normal animals continue to focus on specific immune responses dominated by T cells using restricted sets of antigen receptors. These complementary strategies provide direct access to the dynamics of helper T-cell differentiation in vivo. PMID- 8725953 TI - The use of dominant-negative mutations to elucidate signal transduction pathways in lymphocytes. AB - Recent publications document an exponential increase in the use of dominant negative mutations as tools for the experimental dissection of lymphocyte signaling pathways. This approach may be the only one available for in vitro analysis of cell lines. Moreover, when implemented in transgenic animals, dominant-negative mutations boast certain advantages over gene-targeting strategies. PMID- 8725954 TI - Lymphocyte development. Transcriptional regulation of B- and T-cell development. PMID- 8725955 TI - Immunological techniques. PMID- 8725956 TI - Towards the neuronal correlate of visual awareness. AB - Several encouraging developments towards identifying the neuronal correlate of visual awareness have emerged recently. Increasingly sophisticated behavioral paradigms permit the study of visual awareness in humans as well as in non-human primates. In patients with anatomically restricted lesions in striate and extrastriate cortex, highly informative deficits of visual awareness are observed. Similar deficits can be obtained in normal observers with a novel class of psychophysical displays. Taken together, these results suggest that the contents of visual awareness reflect neuronal activity in certain extrastriate, but not in striate, visual cortical areas. PMID- 8725957 TI - Brain representation of object-centered space. AB - Object-centered spatial awareness underlies many important cognitive functions, including reading, which requires registering the locations of letters relative to a word, and pattern recognition, which requires registering the locations of features relative to a whole pattern. Recent studies have elucidated the nature of the brain mechanisms underlying this form of spatial awareness by showing the attention tends to focus on objects rather than on regions of space: by demonstrating that each hemisphere contributes selectively to awareness of the opposite half of object space, and by revealing that neurons in some cortical areas are selective for particular locations in object space. These results are concordant with the general idea that imagining or attending to an object is accompanied by projecting its image onto a neural map of object-centered space. An important aim for future studies will be to test and extend this 'object map' hypothesis. PMID- 8725958 TI - The binding problem. AB - Perceptual representations depend on distributed neural codes for relaying the parts and properties of objects. Some mechanism is needed to 'bind' the information relating to each object and to distinguish it from others. Possible candidates include cells tuned to conjunctions of features, spatial attention, and synchronized firing across separate but interconnected areas of the brain. Deficits in neurological patients suggest a role for the parietal cortex in the binding process. Several current models combine these ideas. PMID- 8725959 TI - The anatomy, physiology and functions of the perirhinal cortex. AB - The perirhinal cortex is a polymodal association area that contributes importantly to normal recognition memory. A convergence of recent findings from lesion and electrophysiological studies has provided new evidence that this area participates in an even broader range of memory functions than previously thought, including associative memory and emotional memory, as well as consolidation functions. These results are consistent with neuroanatomical research showing that this area has strong and reciprocal connections with widespread cortical sensory areas and with other memory-related structures, including the hippocampal formation and amygdala. PMID- 8725960 TI - Is the rodent hippocampus just for 'place'? AB - The prominent view that the rodent hippocampus is dedicated to spatial memory has been challenged recently by observations that both limit the nature of hippocampal spatial representation and extend its scope beyond literal space. These findings reveal that the rodent hippocampus mediates memory representations on the basis of non-spatial, as well as spatial, relations among items in memory, and supports access to these memories in a variety of situations. Therefore, the defining features of hippocampal representation in rodents, as in humans, lie not in the modality of the information processed, but in the organization of the information that supports a capacity for flexible memory expression. PMID- 8725961 TI - Head direction cells: properties and functional significance. AB - The strong signal carried by head direction cells in the postsubiculum complements the positional signal carried by hippocampal place cells; together, the directional and positional signals provide the information necessary to permit rats to generate and carry out intelligent, efficient solutions to spatial problems. Our opinion is that the hippocampal positional system acts as a cognitive map and that the role of the directional system is to put the map into register with the environment. In this way, paths found using the map can be properly executed. Head direction cells have recently been discovered in parts of the thalamus reciprocally connected with the postsubiculum; such cells provide important clues to the organization of the directional system. PMID- 8725962 TI - Can cognitive neuroscience illuminate the nature of traumatic childhood memories? AB - Recent findings from cognitive neuroscience and cognitive psychology may help explain why recovered memories of trauma are sometimes illusory. In particular, the notion of defective source monitoring has been used to explain a wide range of recently established memory distortions and illusions. Conversely, the results of a number of studies may potentially be relevant to forgetting and recovery of accurate memories, including studies demonstrating reduced hippocampal volume in survivors of sexual abuse, and recovery from functional and organic retrograde amnesia. Other recent findings of interest include the possibility that state dependent memory could be induced by stress-related hormones, new pharmacological models of dissociative states, and evidence for 'repression' in patients with right parietal brain damage. PMID- 8725963 TI - Blindsight revisited. AB - Some human patients with lesions to their primary visual (striate) cortex (V1) demonstrate residual visual capacity, but without acknowledged perceptual awareness. This phenomenon has been termed blindsight. Recent results from work on blindsight patients suggest that it is unlikely to be attributable to intact residual areas (tags) of V1. Previous research has reported that blindsight patients can retain the ability to detect monochromatic light and grating stimuli, and to discriminate orientation and direction of movement in their 'blind' fields. These findings have been joined by reports that these patients also are sensitive to, and are able to discriminate, wavelength in the absence of any experience of 'colour'. This reveals that retinal pathways other than those to the striate cortex are crucially involved in vision. Conditions can be controlled for obtaining either acknowledged awareness or unawareness of discrimination of the direction of a small moving target in blindsight patients. This potentially offers the possibility to determine whether there are structures uniquely involved in visual awareness. Monkeys lacking V1 also clearly demonstrate residual visual capacity, and some evidence exists that they also experience 'blindsight'. PMID- 8725964 TI - The amygdala and emotion. AB - The amygdala complex has long been known as part of the neural circuitry critical for emotion. Beyond its role in emotional reactivity, studies of animal models and patients with amygdala damage demonstrate its importance in emotional learning, whereby cues acquire significance through association with rewarding or aversive events. Although its function in associative learning has become well established, other recent research has advanced the concept that the amygdala regulates additional cognitive processes, such as memory or attention. For example, a correspondence in the function of the amygdala has recently been shown in the modulation of memory in humans and laboratory animals. The use of animal models has progressively defined the circuitry for these functions within the amygdala and its interconnections with other brain systems, including pathways through which the amygdala modulates memory and regulates attention. These various lines of research are progressively advancing our understanding of the amygdala's role in providing linkages between affect and cognition. PMID- 8725965 TI - Neurobehavioural mechanisms of reward and motivation. AB - The analysis of the behavioural and neural mechanisms of reinforcement and motivation has benefited from the recent application of learning theory and better anatomical knowledge of the connectivity of certain key neural structures, such as the nucleus accumbens. This progress has enabled the dissection of motivational processes into components that can begin to be related to the functioning of specific limbic cortical structures that project to different compartments of the ventral striatum. PMID- 8725966 TI - Modulation of memory storage. AB - For several decades, the concept of modulation of memory storage has significantly influenced research investigating neurobiological memory mechanisms. New evidence provides additional support for the view that stress hormones released during emotionally arousing situations modulate memory processes. Recent experiments have investigated the role of sympathetic adrenomedullary hormones in emotional memory in humans, as well as the role of adrenocortical hormones, primarily in animal studies. Further, it is becoming increasingly clear that the sympathetic adrenomedullary and the pituitary adrenocortical systems interact to modulate memory storage. Other new evidence emphasizes the role of peripheral influences to the brain on emotional memory, as well as the critical contribution of the amygdaloid complex in modulation of memory by emotional arousal. PMID- 8725967 TI - Neurobiology of addiction. AB - Addictive drugs have habit-forming actions that can be localized to a variety of brain regions. Recent advances in our understanding of the chemical 'trigger zones' in which individual drugs of abuse initiate their habit-forming actions have revealed that such disparate drugs as heroin, cocaine, nicotine, alcohol, phencyclidine, and cannabis activate common reward circuitry in the brain. Although these drugs have many actions that are distinct, their habit-forming actions (and perhaps the relevant elements of their disparate withdrawal symptoms) appear to have a common denominator, namely, similar effects in the brain mechanisms of reward. PMID- 8725968 TI - Cognitive influences on human autonomic nervous system function. AB - Recent studies of human autonomic nervous system function have challenged the traditional view that the parasympathetic and sympathetic branches have opposite effects. In addition, studies of autonomic activity that accompanies attention, orienting and learning have demonstrated that the autonomic nervous system is not simply a 'non-cognitive' and automatic part of brain function. The autonomic and central nervous systems are intimately related, and new research is beginning to link brain function with autonomic function in a dynamic way. PMID- 8725969 TI - Sex, sexual orientation and sex hormones influence human cognitive function. AB - Superior performance by women on a task requiring object location memory has challenged the traditional view that men excel on all spatial tasks. Sexual orientation is also associated with variation in cognitive ability pattern, but such association appears to be more consistent for a real-world targetting task than for paper-and-pencil spatial tests. Finally, there is increasing evidence that early exposure to sex hormones has lasting effects on problem-solving behaviour; moreover, current fluctuations in sex hormones in both men and women are associated with changes in cognitive pattern. PMID- 8725970 TI - CREB and the formation of long-term memory. AB - Cyclic AMP response element binding protein (CREB)-responsive transcription plays a central role in the formation of long-term memory in Drosophila, Aplysia and mice. Agents that disrupt the activity of CREB specifically block the formation of long-term memory, whereas agents that increase the amount or activity of the transcription factor accelerate the process. These results have led to the recent hypothesis that CREB is pivotal in the switch from short-term (protein synthesis independent) to long-term (protein synthesis dependent) memory. PMID- 8725971 TI - Plasticity in visual perception and physiology. AB - Many factors influence our perception of local features. What we see is not strictly a reflection of the physical characteristics of a scene, but instead is highly dependent on the processes by which our brain attempts to interpret the scene. As a result, our percepts are shaped by the context within which local features are presented, by our previous visual experiences (operating over a wide range of time scales), and by our expectations of what is likely to be before us. The substrate for these influences is found in the lateral interactions operating within individual areas of the cerebral cortex and in the feedback from higher to lower order cortical areas. PMID- 8725972 TI - Cognitive neuroscience. PMID- 8725973 TI - The association between age of hospitalized patients and the delivery of advanced cardiac life support. AB - OBJECTIVE: To determine the extent of variability in the administration of advanced cardiac life support (ACLS) and to determine if age is associated with variability. DESIGN: Retrospective cohort. SETTING: Urban teaching hospital. PATIENTS: One hundred twenty-two adult inpatients without a "do-not resuscitate" order who suffered cardiopulmonary arrest during 1993. MEASUREMENTS AND MAIN RESULTS: Of the total, 35 (29%) survived the arrest and 87 (71%) died. Among the nonsurvivors, two patients received no ACLS and six were not intubated, despite the inclusion of intubation in all ACLS protocols. Of the 87 nonsurvivors, 31 had a single electrocardiographic rhythm during their arrest and should have had similar ACLS trials. However, the 9 nonsurvivors with ventricular fibrillation received a range of 0 to 17 interventions, the 11 with electromechanical dissociation received 1 to 22, and the 11 with asystole received 0 to 14. Based on a protocol-derived definition of a minimal trial of ACLS (a "short ACLS trial") for all 87 nonsurvivors, age greater than 75 was associated with receiving a short trial. Dependent functional status and being on a medical service were also associated with a short ACLS trial. In a logistic regression model including these variables as covariates, age remained significantly associated with a short ACLS trial; odds ratio, 9.71 (95% confidence interval 1.68, 56.1). CONCLUSIONS: Wide variability exists in the administration of ACLS at the studied site. The finding that some patients receive no ACLS suggests that physicians at this site may be making bedside determinations of the likelihood of its benefit based on individual patient characteristics. The association between older age and short ACLS trials among all nonsurvivors suggests that age is most important of these characteristics. PMID- 8725974 TI - Comorbidity as a correlate of length of stay for hospitalized patients with acute chest pain. AB - OBJECTIVE: To determine whether comorbid medical conditions as measured with the Charlson Comorbidity Index are independent correlates of length of stay after adjusting for other clinical and socioeconomic data. DESIGN: Prospective cohort study. SETTING: Urban teaching hospital. PATIENTS: All 1,261 patient aged 30 years or more who were admitted to this hospital after coming to the emergency department with acute chest pain between October 1990 and May 1992. MEASUREMENTS AND OUTCOMES: Clinical data including comorbid medical conditions used in the Charlson index were prospectively recorded by the evaluating physician at the time of admission or by a research nurse who was blinded to the subsequent events. History of myocardial infarction was excluded from the calculation of the Charlson index score. Charlson index scores were 0 to 1 for 921 patients (73%), 2 to 3 for 263 (21%), and greater than 3 for 77 (6%). Unadjusted means (+/- SD) lengths of stay in these groups were 4.4 +/- 5.2, 5.2 +/- 5.9, and 7.5 +/- 9.3 days, respectively. In multiple linear regression analysis, compared with Charlson index scores of 0 to 1, scores of 2 to 3 and greater than 3 were significant (p < .01) independent correlates of the log transformation of length of stay after adjusting for clinical data from the initial presentation and subsequent course (model R2 = .510). In an analysis restricted to the 795 patients without clinical complications, a Charlson index score greater than 3 was an independent correlate of length of stay compared with scores of 0 to 1 (p < .01). Individual comorbid conditions were not significant correlates of length of stay after controlling for Charlson index score. CONCLUSIONS: In this population of patients with acute chest pain, comorbidity as measured with the Charlson index was independently associated with length of stay after adjustment for other clinical data. After adjusting for the Charlson index, no separate comorbid condition was significantly correlated with length of stay. These findings suggest that the Charlson index can be used to adjust for comorbidities in analyses of length of stay for patients with this condition. PMID- 8725975 TI - Primary care and receipt of preventive services. AB - OBJECTIVE: To examine whether health insurance, a regular place of care, and optimal primary care are independently associated with receiving preventive care services. DESIGN: A cross-sectional telephone survey. SETTING: Population based. PARTICIPANTS: Probability sample of 3,846 English-speaking and Spanish-speaking women between the ages of 18 and 64 in urban California. INTERVENTIONS: Women were asked about their demographic characteristics, financial status, health insurance status, need for ongoing care, regular place of care, and receipt of blood pressure screening, clinical breast examinations, mammograms, and Pap smears. Women who reported a regular place of care were asked about four components of primary care: availability, continuity, comprehensiveness, and communication. MEASUREMENTS AND MAIN RESULTS: In multivariate analyses that controlled for differences in demographics, financial status, and need for ongoing care, having a regular place of care was the most important factor associated with receiving preventive care services (p < .0001). Having health insurance (p < .001) and receiving optimal primary care from the regular place of care (p < .01) further significantly increased the likelihood of receiving preventive care services. CONCLUSION: A regular source of care is the single most important factor associated with the receipt of preventive services, but optimal primary care from a regular place increases the likelihood that women will receive preventive care. PMID- 8725976 TI - Physician extenders for cost-effective management of hypercholesterolemia. AB - OBJECTIVE: Treatment of elevated cholesterol levels reduces morbidity and mortality from coronary heart disease in high-risk patients, but can be costly. The purpose of this study was to determine whether physician extenders emphasizing diet modification and, when necessary, effective and inexpensive drug algorithms can provide more cost-effective therapy than conventional care. DESIGN: Randomized controlled trial. SETTING: A Department of Veterans Affairs Medical Center. PATIENTS: Two hundred forty-seven veterans with type IIa hypercholesterolemia. INTERVENTIONS: Patients assigned to either a cholesterol treatment program (CTP) or usual health care provided by general internists (UHC). CTP included intensive dietary therapy administered by a registered dietitian utilizing individual and group counseling and drug therapy initiated by physician extenders for those failing to achieve goal low-density lipoprotein (LDL) levels with diet alone. A drug selection algorithm for CTP subjects utilized niacin as initial therapy followed by bile acid sequestrants and lovastatin. Subjects were followed prospectively for 2 years. MEASUREMENTS: Primary outcome measurements were effectiveness of therapy defined as reductions in LDL cholesterol (LDL-C), and whether goal LDL-C levels were achieved; costs of therapy; and cost-effectiveness defined as the cost per unit reduction in the LDL C. MAIN RESULTS: Total program costs were higher for CTP patients than for UHC patients ($659 +/- $43 vs $477 +/- $42 per patient, p < .001). However, at 24 months the patients in CTP were more likely to achieve LDL goal levels (65% vs 44%, p < .005), and also achieved greater reductions in LDL-C 27% +/- 2% vs 14% +/- 2% at 24 months, p < .001). Program costs per unit (mmol/L) reduction in the LDL-C, a measure of cost-effectiveness, was significantly lower for CTP ($758 +/- $58 vs $1,058 +/- $70, p = .002). CONCLUSIONS: Although more expensive than usual care, the greater effectiveness of physician extenders implementing cholesterol treatment algorithms resulted in more cost-effective therapy. PMID- 8725977 TI - Long-term outcomes of patients receiving percutaneous endoscopic gastrostomy tubes. AB - OBJECTIVE: Percutaneous endoscopic gastrostomy (PEG) tube placement is the preferred method for long-term enteral feeding of patients who are unable to take food by mouth. Despite the widespread acceptance of the procedure, no large-scale study of the long-term outcomes of patients receiving PEG tubes has been reported. The objective of this study was to determine the survival of patients in whom PEG tubes are placed. DESIGN: Retrospective cohort study using data obtained from two computerized databases. SETTING: Department of Veterans Affairs hospitals. PATIENTS: Seven thousand three hundred sixty-nine patients who received a PEG tube in fiscal years 1990 through 1992. RESULTS: For the 7,369 patients, the mean age was 68.1 years and 98.6% were men. PEG tubes were most commonly places in patients with cerebrovascular disease (18.9%), other organic neurologic disease (28.6%), or head and neck cancer (15.7%). Although the complication rate of the procedure itself was low (4%), because of the severity of their underlying disease, 1,732 patients (23.5%) died during the hospitalization in which the PEG tube was placed. The median survival of the full cohort was 7.5 months. CONCLUSIONS: This study documents the widespread placement of PEG tubes in severely ill patients, half of whom are in the terminal phase of their illness. Further study is needed to determine whether these patients benefit from PEG tube placement in terms of their quality of life and survival. PMID- 8725978 TI - The review process used by US health care plans to evaluate new medical technology for coverage. AB - OBJECTIVE: To examine the process and information used by medical directors (MDs) of private health plans to make medical coverage determinations for new medical technologies, and to assess the influence of plan characteristics on the process. DESIGN: Cross-sectional national survey. PARTICIPANTS: Two hundred thirty-one MDs at private health plans representing 66% and 72% of the US population covered by HMOs and indemnity plans, respectively. MEASUREMENTS: Actual and optimal review process, final decision authority, sources, and evidence used for technology coverage decisions. RESULTS: In 96% of plans, MDs take part in the medical policy review process for new technology. However, MDs have final authority over coverage decisions in only 27% of plans. Indemnity plans are more likely to assert that MDs should be responsible for final decisions, odds ration (OR) = 3.3 (95% confidence interval [95% CI] 1.4, 10). Optimal sources of information of new technology were journals, medical society statements or practice guidelines, and opinions of national experts. Actual sources of information used differed from optimal ones; local experts were used more often than is considered optimal (p < .001). For-profit plans were more likely than nonprofit plans to use national experts, OR 2.5 (95% CI 1.3, 5.0), and practice guidelines, OR 5.0 (95% CI 2.5, 10). Randomized trials (94% of MDs) meta-analyses (61%), and reviews (42%) were considered the best evidence for making coverage decisions. Barriers to making optimal decisions were lack of timely evidence on effectiveness and cost effectiveness, not legal or regulatory issues; HMO, small, and nonprofit plans were two or three times more likely to list lack of cost-effectiveness data than their counterparts (p < .05). CONCLUSIONS: Although MDs are nearly always involved in the technology evaluation process, a minority of MDs retain final authority over coverage decisions. Evidence from strong scientific research designs is the most frequently cited basis for decisions, but there is need for more timely, rigorous scientific evidence on medical interventions. How a health plan evaluates a new medical technology for coverage varies with identifiable plan characteristics. PMID- 8725979 TI - How severity measures rate hospitalized patients. PMID- 8725980 TI - Hospitalized patients' satisfaction with caregivers' conduct and physical surroundings. AB - The study examined the relative contribution of caregivers' conduct and physical conditions of the hospital setting to patients' global satisfaction with health care, while controlling for self-assessed health status. The study was performed in the Soroka Medical Center, which is the only inpatient facility for a population of 360,000 in southern Israel. Patients were interviewed using a structured questionnaire. Inclusion criteria were patients over 17 years of age, fluency in Hebrew, and hospitalization in the internal medicine wards for 24 hours or more. The dependent variable was "global satisfaction." The independent variables were "satisfaction with caregivers' conduct," "satisfaction with physical surroundings and conditions," and "self-assessment of health." The three independent variables accounted for 36.8% of the variation in patients' satisfaction with care, of which 87% relate to "satisfaction with caregivers' conduct." In conclusion, hospital patients value the caregivers' conduct more than the physical environment. PMID- 8725981 TI - "Credible clinical" findings as predictors of death. PMID- 8725982 TI - Alcohol withdrawal treatment practices. PMID- 8725983 TI - Pan-European survey of patients' attitudes to antibiotics and antibiotic use. AB - This study was carried out to determine patient perceptions of respiratory tract infections and attitudes to taking antibiotics, thus helping doctors to have a better understanding of their patients and their requirements. Telephone interviews were conducted in the UK, Belgium, France, Italy, Spain and Turkey using standardized questionnaires directed at patients who had taken an antibiotic or given one to their child for a respiratory tract infection within the previous 12 months. Approximately 200 working adults (< or = 55 years), 200 elderly adults (> 55 years) and 200 mothers of children (< 12 years) from each country were contacted; in total, 3610 subjects. Pressure on GPs to prescribe antibiotics was highlighted by over 50% of interviewees' believing that they should be prescribed for most respiratory tract infections. Although interviewees were positive about antibiotics, with over 75% judging them to be effective and to speed recovery, some ambivalence was shown. Most patients waited 2-3 days before consulting their doctor and over 80% of respondents expected symptoms to improve after 3 days' treatment. This provided a natural watershed for compliance, with most defaulters stopping after 3 days because they felt better. Second only to fewer side-effects, patients ranked shorter and more convenient dosage alongside efficacy as the improvements most sought in antibiotic therapy. In conclusion, patients regarded antibiotics as important in the treatment of respiratory tract infections, with interest shown in short-course, once-daily therapy. Doctors, however, need to reassure patients that short courses will eradicate infections and have minimal adverse effects on the immune system. PMID- 8725985 TI - Natural alpha interferon in multiple sclerosis: results of three preliminary series. AB - In three preliminary, sequential series conducted between 1987-1991, natural human leukocyte interferon-alpha [HuIFN(Le)] was evaluated in 49 interferon-naive multiple sclerosis (MS) patients. In this study, safety data are reported for all 49 patients, and efficacy data for 45 of 49, each patient having received 3-12 months of therapy during the initial study year with observation only in the second year. No major side-effects for HuIFN(Le) were observed at doses of 5-30 million international units (MIU) per week for 3-12 months, although severe fatigue caused some patient drop-outs prior to completion of 3 months of therapy. The relapse rate in patients having received HuIFN(Le), prospectively evaluated in two series (34 patients), was reduced by 80% at 2 years. An unexpected trend towards improved disability provoked a retrospective analysis. In the first year, 36 of 45 (80%) patients improved or stabilized (P = 0.0001); of these 26 of 45 (58%) improved and 10 of 45 (22%) stabilized (P = 0.0023, chi 2 = 9.3). In year 2, 34 of 45 (76%) were improved/stabilized (P = 0.001); of these, 24 of 45 (53%) remained improved and 10 of 45 (22%) remained stable (P = 0.01, chi 2 = 6.6). A trend to greater efficacy at the higher doses for longer periods of therapy, up to 12 months, was observed. PMID- 8725984 TI - Serum interleukin 2 and soluble interleukin 2 receptor in chronic active hepatitis C: effect of interferon therapy. AB - T lymphocytes produce interleukin 2 (IL-2) associated with the expression of a soluble receptor for IL-2 (sIL-2R) on the surface of the cells, and these cytokines may contribute to hepatic injury in chronic active hepatitis C (CAH-C). Serum IL-2 and sIL-2R levels were analysed by enzyme linked immunosorbent assay in 22 patients (eight female, 14 male, mean age 42.6 years) with CAH-C. Eight patients had been given interferon alpha-2a (IFN alpha-2a), 3 million international units, three times weekly, for a mean of 12 weeks while the others were not on treatment. Serum IL-2 levels were 60.8 +/- 9.5 pg/ml, 66.6 +/- 5.7 pg/ml and 59.1 +/- 4.0 pg/ml in the treated patients, untreated patients and controls, respectively. Serum sIL-2R levels were 1631 +/- 194 pg/ml, 4016 +/- 1076 pg/ml and 1169 +/- 115 pg/ml in treated patients, untreated patients and controls, respectively. There were no significant differences in serum IL-2 levels between the groups (P > 0.05) while a significant difference was found in serum sIL-2R levels between untreated patients and controls (P = 0.0032). Serum sIL-2R levels were lower in patients treated with IFN alpha-2a than in untreated patients but this difference was not statistically significant. This preliminary study indicates that there are no significant changes in serum IL-2 levels in CAH C patients, but that sIL-2R concentrations are raised in untreated patients though not in treated patients. High serum sIL-2R concentrations may have a role in the pathogenesis of CAH-C. PMID- 8725986 TI - Serum levels of soluble intercellular adhesion molecule-1 and soluble vascular cell adhesion molecule-1 in liver disease, and their changes by treatment with interferon. AB - Serum levels of soluble intercellular adhesion molecule-1 (sICAM-1) and soluble vascular cell adhesion molecule-1 (sVCAM-1) were measured by an enzyme-linked immunosorbent assay in patients with chronic hepatitis (n = 57), liver cirrhosis (n = 19) and hepatocellular carcinoma (n = 33). Serum levels of sICAM-1 and sVCAM 1 were significantly higher in liver disease than those in controls (P < 0.0001 and P < 0.0005, respectively). A total of 22 patients with chronic hepatitis C were treated with interferon. Pretreatment levels of sICAM-1 and sVCAM-1 were not significantly different between complete responders and non-responders. In complete responders, serum sICAM-1 and sVCAM-1 levels 1 year after interferon treatment significantly decreased compared to the pretreatment levels (P < 0.005 and P < 0.05, respectively). Post-treatment levels of sICAM-1 and sVCAM-1 in complete responders were also significantly lower than those in non-responders (P < 0.005 and P < 0.05, respectively). This suggests that monitoring soluble adhesion molecules might be useful in the follow-up of patients with liver disease. PMID- 8725987 TI - Anti-emetic efficacy of tropisetron and metoclopramide. AB - Post-operative vomiting, especially in ambulatory surgical patients, remains a troublesome problem. This placebo-controlled, randomized, prospective double blind trial was designed to evaluate the efficacy of two prophylactic anti-emetic regimens on post-operative vomiting in 1-day thyroid surgery. Altogether 60 elective surgical patients were followed for 4 h post-operatively. All patients were American Society of Anesthesiologists physical status of I or II and aged between 22 and 60 years: group 1 was saline control; in groups 2 and 3, metoclopramide (0.2 mg/kg) or tropisetron (5 mg) was administered, respectively, as an intravenous single dose during induction. Patients were pre-medicated. A standardized anaesthetic technique consisting of thiopentone-succinylcholine for induction and fentanyl-nitrous oxide-halothane-pancuronium for maintenance of anaesthesia was used. A 'rescue' anti-emetic was provided in case of continued vomiting or at the patient's request. Anti-emetic inefficacy was defined as request for rescue anti-emetic and/or vomiting episode during the first 4 h post operation. The number of patients vomiting was 12/20 (60%), 10/20 (50%) and 1/20 (5%) within the first 2 h post-operation in groups 1, 2 and 3, respectively (P > 0.05 for groups 1 and 2; P < 0.01 for groups 2 and 3; P < 0.001 for groups 1 and 3). In group 2, three patients required rescue medication during the first 2 h post-operation, but no significant difference was observed between groups 2 and 3 (P > 0.05). None of the cases in any of the groups needed any rescue medication during post-operative 2-4 h. It is concluded that tropisetron is a highly effective anti-emetic drug in the prophylaxis of post-operative vomiting. PMID- 8725988 TI - Effects of bromocriptine on dopamine turnover with or without levodopa. AB - Bromocriptine, a dopamine agonist, alleviates symptoms of Parkinson's disease, even when administered alone, and is used for its treatment. Better therapeutic effects are, however, achieved when bromocriptine is used in combination with levodopa. In this study, we examined the biochemical changes caused by bromocriptine administration with and without levodopa, and evaluated the effects of the treatments on dopamine turnover in the mouse striatum. Results show that dopamine turnover is suppressed by the administration of bromocriptine alone with a slight decrease in the amount of dopamine, and dopamine turnover is very strongly promoted by the administration of levodopa. When the two drugs are administered together, bromocriptine enhances the levodopa-induced increase in dopamine turnover in the striatum. These findings indicate that bromocriptine therapy in combination with levodopa enhances the dopaminergic function and suggest that the combination therapy of bromocriptine and levodopa shows good efficacy. The results of this study may, thus, provide a theoretical basis for the combination therapy of bromocriptine and levodopa. PMID- 8725989 TI - Relationship between adrenal cortex pathology in Cushing's syndrome and its response to the dexamethasone suppression test. AB - Cushing's syndrome is a severely disabling condition which can cause death if left untreated. Endogenous Cushing's syndrome can be ACTH-dependent or ACTH independent. The ACTH-dependent type is more common and is usually caused by diffuse hyperplasia on the adrenal cortex. This study investigated the response to low- and high-dose dexamethasone suppression testing of 30 adrenalectomized patients with Cushing's syndrome, average age 37.3 +/- 9.7 years. Twenty-four (79.3%) patients were female, and six (20.7%) were male. Bilateral adrenalectomy was performed in 14 (48.2%) patients and unilateral adrenalectomy (nine and seven right adrenalectomy) in 16 (51.8%). Two of the bilateral adrenalectomies were applied via endoscopic surgical approach. In the histopathological evaluation, diffuse hyperplasia was diagnosed in 13 (44.8%) patients and nodular hyperplasia in eight (26.6%), three macronodular and five micronodular hyperplasia. Adrenal cell adenoma was diagnosed in nine (28.6%) patients. Classic dexamethasone suppression testing was performed on all patients. Plasma levels of cortisol were not significantly decreased after low-dose testing, but plasma levels of cortisol were significantly decreased after high-dose testing in the diffuse hyperplasia group. In summary, due to the pathological changes of the adrenal cortex, dexamethasone suppression testing can differentiate between the two types of Cushing's syndrome. PMID- 8725990 TI - Partial effective time (teffp7): a new spirometric parameter for lung function assessment. AB - A new spirometric parameter for lung function assessment, the effective time at the 60-70% part of the forced vital capacity of the lung (teffp7), was considered. A total of 79 subjects were studied in two groups, A and B. In group A (5 healthy non-smokers, 22 patients with obstructive lung disease and 22 with pulmonary fibrosis), forced exhalation and the Helium closed-circuit technique were performed. From these tests, a number of lung function parameters were calculated, including teffp7. In group B (30 patients with bronchial asthma), the same tests were carried out before and 15 min after the administration of two puffs of a bronchodiluent, a beta 2-stimulant. From these tests, various lung function measures were again calculated, as well as the change in these parameters after bronchodilatation. The study shows that teffp7 is a more sensitive index than all the other parameters considered for determining airways condition, either in healthy subjects or in those with airways obstruction. In patients with pulmonary fibrosis, the other parameters considered are also less sensitive than teffp7. From the study of the change after bronchodilatation in patients with bronchial asthma, change in teffp7 was greater than in any other parameter. As teffp7 was the more affected parameter, it was also the more sensitive index for diagnosis of airways obstruction. In conclusion, teffp7 is a sensitive index for the diagnosis of airways obstruction or lung restriction, and it is markedly changed after bronchodilatation. PMID- 8725991 TI - Efficacy and safety of risperidone in psychotic patients: an open study. AB - This open prospective study was undertaken to determine the efficacy and safety of a fixed dose (6 mg) of risperidone in psychotic patients. Hospital in-patients who fulfilled DSM-111-R criteria for schizophrenia, schizoaffective and bipolar disorders were eligible for entry into the study (n = 15). Patients who were on other antipsychotics had a washout period of 1 week before they were started on the drug. A fixed dose of risperidone was administered (6 mg). The Brief Psychiatric Rating Scale (BPRS), Negative Symptom Rating Scale (NSRS) and Abnormal Involuntary Movement Scale were used to measure psychopathology and extrapyramidal side-effects. Five patients dropped out of the study. Two patients became very agitated and potentially aggressive, one patient became very restless and did not respond to benzodiazepines, and one dropped out because of restlessness that did not respond to clonazepam. Of the 10 patients who completed the study, 50 per cent reduction on BPRS and NSRS was achieved by five and six patients respectively. There was a marginally significant trend towards a greater reduction in the magnitude of negative symptoms. Four patients required treatment with anticholinergic drugs. Risperidone was effective in resistent psychotic patients, but agitated and impulsive psychotic patients with positive symptoms may not be best candidates for treatment with risperidone. On average, negative symptoms respond better than positive symptoms. PMID- 8725992 TI - Drug sensitivity pattern of E. coli causing urinary tract infection in diabetic and non-diabetic patients. AB - Escherichia coli was isolated from 65 urine samples out of 300 specimens collected from diabetic and non-diabetic patients with urinary tract infections. All isolates were screened for susceptibility to commonly used antibiotics. No remarkable difference in sensitivity pattern was observed between the E. coli isolates from diabetic and non-diabetic out-patients. However, the isolates from hospitalized diabetic patients showed a clear difference in sensitivity; E. coli from this group was more resistant to commonly used antibiotics compared with those from non-hospitalized diabetic and non-diabetic patients. PMID- 8725993 TI - Effects of a short course of treatment with acebrophylline on the mucus rheological characteristics and respiratory function parameters in patients suffering from chronic obstructive pulmonary disease. AB - A total of 30 patients (27 males and 3 females) with a mean age of 62.6 +/- 3.9 years, suffering from chronic obstructive pulmonary disease were recruited into this open study to evaluate the clinical efficacy and the safety of a short course of treatment with 100 mg acebrophylline, twice daily for 14 days. To assess the effectiveness of the drug, symptoms and signs, such as cough intensity and frequency, auscultatory pattern, dyspnoea, cyanosis, difficulty of expectoration, sputum quantity, appearance and density, were evaluated at baseline and after 1, 3, 5, 7, 10 and 14 days of therapy. In addition the rheological properties of the bronchial mucus (viscosity and spinnability) were measured and respiratory function tests were performed before and after treatment. There was a progressive improvement of all evaluated symptoms and signs, resulting in improvement of the auscultatory pattern, as well as dyspnoea. This was accompanied by improvement or normalization of the respiratory function indices, which showed statistically significant differences (P < 0.01) between the baseline and the endpoint values, apart from total lung capacity. Blood-gas analysis demonstrated a significant increase of PaO2 and a significant decrease of PaCO2 values at the end of the treatment period (both P < 0.01). In general, acebrophylline was well tolerated. No clinically relevant or significant changes in any of the routine laboratory parameters were found on comparing the values obtained before and after treatment. Only three patients complained of epigastric pain, but this was not so severe or long-lasting as to require the discontinuation of the treatment. PMID- 8725994 TI - Safety and efficacy of picotamide, a dual anti-thromboxane agent, in patients with thrombocytosis and a previous thromboembolic event: a 1-year observational study. AB - Patients with chronic myeloproliferative disease are at increased risk of both thromboembolic and haemorrhagic complications. Cerebral thrombosis is a common cause of death in myeloproliferative disease patients. Picotamide is a new anti platelet drug sharing a dual anti-thromboxane activity: inhibition of thromboxane A2 synthase and thromboxane A2 receptor antagonism. Picotamide inhibits in vitro and ex vivo platelet aggregation induced by different agonists. Interestingly, in vitro studies show that picotamide is able to increase prostacycline biosynthesis. In the clinical setting, picotamide treatment induces only a slight prolongation of bleeding time. The safety and efficacy of picotamide long-term treatment in 15 patients with essential thrombocytosis and a positive history of previous thromboembolic events was evaluated. After 12-month treatment with picotamide no patients suffered from thrombotic events and only one minor and transient bleeding episode was observed. This observational long-term trial shows that picotamide treatment in patients with thrombocytosis at high risk of thrombotic events is safe and well tolerated. Picotamide did not increase the risk of bleeding in these patients, while at the same time, no thrombotic events were observed during the 1-year treatment. PMID- 8725995 TI - Didanosine measurement by radioimmunoassay. AB - Didanosine is commonly prescribed as monotherapy or as part of a combination regimen for patients with human immunodeficiency virus infection. The use of lower doses, either as part of a combination regimen or as a result of dose reduction secondary to clinical intolerance, requires that a sensitive assay method be available for either traditional or population-based pharmacokinetic evaluations. We evaluated a radioimmunoassay technique with a standard curve range of 0 to 100 ng/ml in human plasma, urine, and cerebrospinal fluid and assessed its accuracy and precision for use in pharmacokinetic studies. PMID- 8725996 TI - Rifapentine is active in vitro and in vivo against Toxoplasma gondii. AB - Rifapentine, a derivative of rifamycin, was examined for its in vitro and in vivo activities against the protozoan parasite Toxoplasma gondii. The drug inhibited the intracellular replication of parasites and was not cytotoxic for the host cells at inhibitory concentrations. Mice infected either intraperitoneally with tachyzoites of the RH strain or orally with tissue cysts of the C56 strain were protected against death by treatment with rifapentine. The degree of protection was similar to that induced by atovaquone and apparently higher than that induced by rifabutin. Rifapentine may be a useful drug for the treatment of toxoplasmosis in immunocompromised individuals. PMID- 8725997 TI - Electronic compliance assessment of antifungal prophylaxis for human immunodeficiency virus-infected women. AB - Several prophylactic medications for opportunistic or recurrent infections are used in human immunodeficiency virus-infected individuals. Essential to the efficacy evaluation of these agents is the accurate reporting of medication compliance. We hypothesized that poor patient compliance with thrice-weekly fluconazole prophylaxis would correlate with the occurrence of clinical events. Fluconazole compliance was monitored electronically by using the Medication Event Monitoring Systems with 19 women receiving fluconazole at 50 mg thrice weekly for prophylaxis of recurrent mucocutaneous candidiasis. During 202 patient-months of follow-up, eight breakthrough episodes of mucocutaneous candidiasis developed in four women; compliance data were available for seven of these episodes. At 6 months of therapy, more women with greater than or equal to 80% compliance were disease free compared with women with less than 80% compliance (P < 0.05; the Fisher exact test). These data suggest that documentation of medication compliance is essential in studies of chronic prophylaxis in human immunodeficiency virus-infected patients to properly evaluate drug efficacy and to avoid erroneous conclusions concerning drug failure. PMID- 8725998 TI - Comparative resistance of Candida albicans clinical isolates to fluconazole and itraconazole in vitro and in vivo in a murine model. AB - Relationships between azole susceptibility and in vivo response to antifungal therapy in a murine model of candidiasis were investigated for Candida albicans isolates sampled from human immunodeficiency virus type 1-positive patients with oropharyngeal candidiasis. The susceptibilities of seven clinical isolates and two reference strains to fluconazole (FCZ) and itraconazole (ITZ) were determined in vitro by the broth microdilution method. Four isolates were resistant to FCZ and ITZ, two were susceptible to both azoles, and three were resistant to FCZ and susceptible to ITZ (dissociated resistance). CD1 mice were inoculated with each isolate and treated with either FCZ or ITZ (drug regimen, 5 mg/kg of body weight twice daily for 5 days). Quantitative cultures of kidneys were performed at the end of the treatment. On the other hand, the survival rates of the mice were followed daily. These two parameters were clearly correlated with in vitro susceptibility. Thus, the phenomenon of a dissociation of resistance to FCZ and ITZ may be found in vivo as well as in vitro. PMID- 8725999 TI - Evaluation of reverse transcriptase and protease inhibitors in two-drug combinations against human immunodeficiency virus replication. AB - Current treatments for human immunodeficiency virus (HIV) include both reverse transcriptase and protease inhibitors. Results from in vitro and clinical studies suggest that combination therapy can be more effective than single drugs in reducing viral burden. To evaluate compounds for combination therapy, stavudine (d4T), didanosine (ddI), or BMS-186,318, an HIV protease inhibitor, were combined with other clinically relevant compounds and tested in a T-cell line (CEM-SS) that was infected with HIV-RF or in peripheral blood mononuclear cells infected with a clinical HIV isolate. The combined drug effects were analyzed by the methods described by Chou and Talalay (Adv. Enzyme Regul. 22:27-55, 1984) as well as by Prichard et al. (Antimicrob. Agents Chemother. 37:540-545, 1993). The results showed that combining two nucleoside analogs (d4T-ddI, d4T-zidovudine [AZT], and d4T-zalcitabine [ddC]), two HIV protease inhibitors (BMS-186,318 saquinavir, BMS-186,318-SC-52151, and BMS-186,318-MK-639) or a reverse transcriptase and a protease inhibitor (BMS-186,318-d4T, BMS-186,318-ddI, BMS 186,318-AZT, d4T-saquinavir, d4T-MK-639, and ddI-MK-639) yielded additive to synergistic antiviral effects. In general, analysis of data by either method gave consistent results. In addition, combined antiviral treatments involving nucleoside analogs gave slightly different outcomes in the two cell types, presumably because of a difference in phosphorylation patterns. Importantly, no strong antagonism was observed with the drug combinations studied. These data should provide useful information for the design of clinical trials of combined chemotherapy. PMID- 8726000 TI - Paradoxical effect of clindamycin in experimental, acute toxoplasmosis in cats. AB - Cats were experimentally inoculated parenterally with the ME49 strain of Toxoplasma gondii to characterize the efficacies of two different dosages of orally administered clindamycin hydrochloride in the treatment of ocular toxoplasmosis. Concentrations of clindamycin hydrochloride at levels previously suggested to be inhibitory to T. gondii replication in vitro were achieved in the serum and aqueous humor but not in the cerebrospinal fluid. Antibiotic therapy, initiated 7 days after inoculation, resulted in no significant difference in the morphometric severity of ocular posterior segment lesions compared with that in the control groups. Treatment appeared to blunt T. gondii-specific immunoglobulin M production but had no significant effect on immunoglobulin G titers. Paradoxically, clindamycin administration was associated with increased morbidity and mortality from hepatitis and interstitial pneumonia, which are characteristic of generalized toxoplasmosis. Serum tumor necrosis factor alpha activity was detected at moderate levels in all groups of cats and correlated with the severity of clinical disease. The results of the study suggest that clindamycin, when administered at this specific time interval following inoculation, does not ameliorate ocular lesions and has a detrimental effect on the clinical course of acute, experimental toxoplasmosis in cats. The factors responsible for and the relevance of this detrimental effect to naturally occurring toxoplasmosis in humans and pet cats were not clear from the study but may relate to an antibiotic associated decrease in the antitoxoplasmic activity of phagocytic cells responsible for the control of T. gondii. PMID- 8726001 TI - Pyrimethamine pharmacokinetics in human immunodeficiency virus-positive patients seropositive for Toxoplasma gondii. AB - Pyrimethamine pharmacokinetics were studied in 11 human immunodeficiency virus (HIV)-positive patients who were seropositive for exposure to Toxoplasma gondii and were taking zidovudine (AIDS Clinical Trials Group Protocol 102). Pyrimethamine was administered at 50 mg daily for 3 weeks to achieve steady state, and pharmacokinetic profiles were determined after administration of the last dose. Noncompartmental and compartmental analyses were performed. Population pharmacokinetic analysis assuming a one-compartment model yielded the following estimates: area under the 24-h concentration-time curve, 42.7 +/- 12.3 micrograms.h/ml; halflife, 139 +/- 34 h; clearance, 1.28 +/- 0.41 liters/h; volume of distribution, 246 +/- 641; and absorption rate constant, 1.5 +/- 1.3 liters/h. These values are similar to those seen in subjects without HIV infection. Pyrimethamine pharmacokinetics did not differ significantly in those subjects who were intravenous drug users. Adverse effects were noted in 73% of those initially enrolled in this study, leading to discontinuation for 38%. No association was noted between pyrimethamine levels and the incidence of adverse events. No significant differences were seen in zidovudine pharmacokinetic parameters obtained from studies performed before and during treatment with pyrimethamine. In summary, pyrimethamine exhibited pharmacokinetics in HIV infected patients that were similar to those in non-HIV-infected subjects and it did not alter the pharmacokinetics of zidovudine in these patients. PMID- 8726002 TI - Modulatory effect of antibiotics on cytokine production by human monocytes in vitro. AB - Some antimicrobial agents have been reported to modify the host immune and inflammatory responses both in vivo and in vitro. Fosfomycin (FOM) and clarithromycin (CAM) have immunomodulatory activity on human lymphocyte function. In the present study, we examined the effects of FOM and CAM on cytokine synthesis by lipopolysaccharide (LPS)-stimulated human monocytes in comparison with that of dexamethasone in vitro. The three drugs demonstrated positive or negative effects on the synthesis of various cytokines by LPS-primed monocytes. They suppressed the synthesis of tumor necrosis factor alpha, interleukin 1 alpha (IL-1 alpha), IL-1 beta, the IL-1 receptor antagonist, and granulocyte-macrophage colony-stimulating factor in a concentration-dependent manner at concentrations between 1.6 and 40 micrograms/ml. On the contrary, the drugs showed different actions on the synthesis of IL-6 and IL-10. Namely, FOM enhanced both IL-6 and IL 10 synthesis, CAM enhanced only IL-10 synthesis, but dexamethasone deeply suppressed the synthesis of both cytokines. These data indicate that antibacterial agents may modify acute-phase inflammatory responses through their effects on cytokine synthesis by monocytes. PMID- 8726003 TI - 6,7-disubstituted 2,4-diaminopteridines: novel inhibitors of Pneumocystis carinii and Toxoplasma gondii dihydrofolate reductase. AB - Four novel, disubstituted diaminopteridines have been identified which antagonize the uptake of a folate precursor (para-aminobenzoic acid) by rat-derived Pneumocystis carinii maintained in short-term axenic culture at concentrations ranging from 4.5 to 26 microM. The compounds were at least 10 to 100 times more active than trimethoprim in this assay. None of these entities exhibited toxicity to mammalian cell lines at < 100 microM. The same structures also caused significant inhibition of Toxoplasma gondii tachyzoite replication within Madin Darby bovine kidney cells at concentrations ranging from 0.1 to 10 microM. Three of the structures (GR92754, AH10639, and AH2504) were at least an order of magnitude more potent than the standard anti-T. gondii agent, pyrimethamine. All three entities were also significantly more potent and selective than pyrimethamine as inhibitors of T. gondii dihydrofolate reductase (DHFR), with 50% inhibitory concentrations within the range of 0.018 to 0.033 microM. One of these compounds, 6,7-dibutyl-2,4-diaminopteridine (GR92754), was also a potent and selective inhibitor of P. carinii DHFR (50% inhibitory concentration, 0.082 microM). GR92754 is the first DHFR inhibitor described that exhibits greater potency, selectivity, and intracellular activity against both organisms than any of the DHFR agents used clinically, namely, trimethoprim, pyrimethamine, and trimetrexate. This information could provide the starting point for examination of the pharmacokinetic and therapeutic potential of GR92754 and related chemical entities with animal models. PMID- 8726004 TI - Antibacterial activities of epiroprim, a new dihydrofolate reductase inhibitor, alone and in combination with dapsone. AB - Epiroprim (EPM; Ro 11-8958) is a new selective inhibitor of microbial dihydrofolate reductase. EPM displayed excellent activity against staphylococci, enterococci, pneumococci, and streptococci which was considerably better than that of trimethoprim (TMP). EPM was also active against TMP-resistant strains, although the MICs were still relatively high. Its combination with dapsone (DDS) was synergistic and showed as in vitro activity superior to that of the TMP combination with sulfamethoxazole (SMZ). The EPM-DDS (ratio, 1:19) combination inhibited more than 90% of all important gram-positive pathogens at a concentration of 2 + 38 micrograms/ml. Only a few highly TMP-resistant staphylococci and enterococci were not inhibited. EPM was also more active than TMP against Moraxella catarrhalis, Neisseria meningitidis, and Bacteroides spp., but it was less active than TMP against all other gram-negative bacteria tested. Atypical mycobacteria were poorly susceptible to EPM, but the combination with DDS was synergistic and active at concentrations most probably achievable in biological fluids (MICs from 0.25 +/- 4.75 to 4 + 76 micrograms/ml). EPM and the EPM-DDS combination were also highly active against experimental staphylococcal infections in a mouse septicemia model. The combination EPM-DDS has previously been shown to exhibit activity in Pneumocystis carinii and Toxoplasma models and, as shown in the present study, also shows good activity against a broad range of bacteria including many strains resistant to TMP and TMP-SMZ. PMID- 8726005 TI - Comparison of D0870, a new triazole antifungal agent, to fluconazole for inhibition of Candida albicans cytochrome P-450 by using in vitro assays. AB - D0870 was 12 to 15 times more active than fluconazole in experiments to determine the MIC for growth arrest for two isolates of Candida albicans. A biochemical comparison of in vitro sterol biosynthesis in cell extracts showed only a twofold superiority of D0870 over fluconazole. A large differentiation (10-fold) in 50% saturating concentrations obtained by examining the binding of the azoles to microsomal P-450 was observed in a type II binding spectrophotometric assay, possibly reflecting the differential affinity for more than one P-450 enzyme. Additional mechanisms besides affinity for the target enzyme sterol 14 alpha demethylase, such as differential intracellular accumulation of drug, may contribute to the differences in antifungal activity. PMID- 8726006 TI - In vitro characterization of aminoglycoside adaptive resistance in Pseudomonas aeruginosa. AB - Aminoglycoside adaptive resistance was characterized in one reference strain and four clinical isolates of Pseudomonas aeruginosa. Adaptive resistance was initiated with a 2-h gentamicin or tobramycin exposure at the MIC. Each P. aeruginosa strain demonstrated an adaptive-resistance period of between 8 and 12 h when tested with both aminoglycosides. Aminoglycoside adaptive resistance was shown to correlate with a decrease in [3H] gentamicin accumulation and a small (5%) but significant (P < 0.05) reduction in proton motive force. The mean generation time of P. aeruginosa during peak levels of adaptive resistance (i.e., maximum reductions in aminoglycoside killing) was not significantly different from that of control organisms (P < 0.05). No changes in outer membrane protein or lipopolysaccharide sodium dodecyl sulfate-polyacrylamide gel electrophoresis profiles were noted when control, adaptively resistant, and postadaptively resistant cells were compared. Cytoplasmic membrane profiles of adaptively resistant cells, however, demonstrated several band changes when compared with control and postadaptively resistant cells. We conclude that the decrease in aminoglycoside accumulation associated with adaptive resistance in P. aeruginosa may be, in part, a function of reductions in proton motive force and/or cytoplasmic membrane protein changes. However, the importance of these changes requires further investigation. PMID- 8726007 TI - Penetration of ceftibuten into middle ear fluid. AB - The penetration of ceftibuten, an extended-spectrum oral cephalosporin, into middle ear fluid (MEF) was evaluated in pediatric patients during a course of daily oral doses of 9 mg/kg of body weight for 10 days. Plasma and MEF collected at 2, 4, 6, or 12 h after at least 3 days of dosing were analyzed for ceftibuten by a high-pressure liquid chromatography method, and the data were used to calculate pharmacokinetic parameters. Plasma and MEF had almost identical maximum concentrations (Cmax) of ceftibuten (14 micrograms/ml). These Cmax values in MEF during acute otitis media were well in excess of the MIC for 90% of the isolates of each of four major pathogens in this disease. The time to Cmax was longer in MEF (4 h) than in plasma (2 h). Excellent penetration (71%) of ceftibuten into MEF was observed on the basis of the area under the curve ratio (MEF/plasma). These data clearly indicate that ceftibuten penetrated well into the MEF to yield clinically effective concentrations. PMID- 8726008 TI - Lack of effect of concomitant zidovudine on rifabutin kinetics in patients with AIDS-related complex. AB - The effect of concomitant dosing with the antiretroviral agent zidovudine (ZDV) on the pharmacokinetics of rifabutin (RBT) was investigated under steady-state conditions. Sixteen human immunodeficiency virus-positive patients with AIDS related complex who had been maintained on stable ZDV therapy for > or = 6 weeks were administered RBT concomitantly for 12 days. Eight patients received daily doses of 300 or 450 mg of RBT. Administration of ZDV was discontinued on day 13, and RBT was given alone for 3 additional days. Four patients receiving 450 mg of RBT discontinued treatment. Under steady-state ZDV and RBT dosing, safety and kinetics assessments were performed on day 13 (ZDV plus RBT) and day 16 (RBT alone). Kinetics on days 13 and 16 demonstrated that RBT (300 or 450 mg) was readily absorbed, with the time at which the plasma concentration was maximal (Tmax) ranging between 2.6 and 2.9 h. At these two doses, the mean steady-state maximal plasma concentrations (Cmax) were 250 and 430 ng/ml on day 13 and 245 and 458 ng/ml on day 16, respectively. RBT kinetics at the two doses were proportional and similar on the basis of estimates of the ratios of the areas under the concentration-time curves over the dosing interval from 0 to 24 h (AUC0 24) (450 mg/300 mg), which were 1.5 and 1.4 for days 13 and 16, respectively. No significant differences were apparent in the mean oral clearance (CLs/F) estimates (range, 1.60 to 1.77 liters/h/kg), which were dose independent and similar for the 2 assessment days, as was the urinary recovery of RBT and its 25 deacetyl metabolite. Low urinary recovery of 25-deacetyl RBT and an AUC metabolite/parent ratio of 0.1 suggest that there is minimal metabolism of RBT via the deacetylation pathway. For RBT, pooled mean (95% confidence interval) ratio (day 13/day 16) estimates for Cmax, Tmax, AUC0-24, and CLs/F were 1.07 (range, 0.77 to 1.38), 1.08 (0.89 to 1.27), 0.97 (0.82 to 1.13), and 1.09 (0.92 to 1.26), respectively. In addition, no significant changes in any of the major safety parameters were detected throughout the study. Therefore, it is concluded that coadministration of ZDV and RBT does not affect the pharmacokinetics and/or safety of RBT in human immunodeficiency virus-positive patients. PMID- 8726009 TI - Variation in postantibiotic effect of clindamycin against clinical isolates of Staphylococcus aureus and implications for dosing of patients with osteomyelitis. AB - Initial measurements of postantibiotic effect (PAE) were made by a standard laboratory method (exposure to 1 mg of clindamycin per liter for 1 h). The range of PAE for 21 strains of Staphylococcus aureus isolated from osteomyelitis patients was 0.4 to 3.9 h, which markedly exceeded the coefficient of variation for the method (6 to 19%). Exposure of S. aureus to three doses of clindamycin at 8-h intervals had no consistent effect on either PAE or MIC. The PAE was dependent on both concentration and duration of exposure to clindamycin: for example, the PAEs for one strain were 1.7 h after exposure to 1 mg/liter for 1 h, 2.4 h after exposure to 4 mg/liter for 1 h, and 5.9 h after exposure to 4 mg/liter for 3 h. Pharmacokinetic simulations showed that the dose required to maintain free serum clindamycin concentrations above the MIC was 300 mg 6 hourly after oral administration (95% confidence interval, 243 to 301 mg) and 1.2 g 6 hourly (95% confidence interval, 305 to 1,145 mg) after intravenous (i.v.) administration. The duration of PAE would have to be at least 2.4 h to allow an increase in the oral dose interval to 8 h or to allow i.v. administration of 300 mg 6 hourly. Additional PAE experiments were performed with the three strains for which PAEs are the shortest after exposure to 1 mg/liter for 1 h (0.4 to 1.2 h). The PAE for these three strains increased markedly to 4.4 to 6.7 h following exposure to 2 mg/liter for 6 h (to mimic the area under the concentration-time curve from 0 to 6 h after a 300-mg dose). These data suggest that oral clindamycin could be administered at 300 mg 8 hourly in the treatment of S. aureus infection, whereas the i.v. dose interval should be 6 h. These suggestions should be confirmed by performing clinical trials. PMID- 8726010 TI - Potentiation of the antimalarial agent rufigallol. AB - We have discovered a remarkable synergistic antimalarial interaction between rufigallol and the structurally similar compound exifone. The synergistic effects were produced in chloroquine-susceptible and chloroquine-resistant clones of Plasmodium falciparum. The degree of potentiation as estimated by standard isobolar analysis was approximately 60-fold for experiments initiated with asynchronous parasites. The most pronounced synergism was observed in experiments with synchronized trophozoite-infected erythrocytes, in which the degree of synergy was at least 300-fold. While the mechanism underlying this drug potentiation remains unresolved, it is hypothesized that rufigallol acts in pro oxidant fashion to produce oxygen radicals inside parasitized erythrocytes. These radicals would attack exifone, thereby initiating its transformation into a more potent compound, a xanthone. PMID- 8726011 TI - In vitro activities of quinolones, beta-lactams, tobramycin, and trimethoprim sulfamethoxazole against nonfermentative gram-negative bacilli. AB - From 1991 to 1995, 8,975 nonfermentative gram-negative bacilli were isolated from patients at The Ohio State University Medical Center: 71% Pseudomonas aeruginosa, 14% Stenotrophomonas maltophilia, 7.6% Acinetobacter baumannii, and < 2% each of 25 other species. The MICs of trovafloxacin (CP-99,219), ciprofloxacin, ofloxacin, ampicillin-sulbactam, piperacillin, piperacillin-tazobactam, ceftazidime, cefoperazone, ceftriaxone, imipenem, tobramycin, and trimethoprim sulfamethoxazole (TMP-SMZ) were determined for 308 isolates, representing 13 species, by a standardized broth microdilution method. The activities of all drugs were species dependent. The fluoroquinolones had inconsistent activity against most species, although several relatively uncommon nonfermenters were consistently susceptible or resistant. Trovafloxacin was considerably more active than ciprofloxacin and ofloxacin against S. maltophilia, A. baumannii, and several less common species. Among the beta-lactams, relative activities varied considerably; overall, imipenem had the broadest spectrum of activity but was inactive against S. maltophilia and Burkholderia cepacia isolates. Tobramycin and TMP-SMZ had stereotypic spectra of activity. Tobramycin was active against most species except S. maltophilia, Alcaligenes xylosoxidans subsp. xylosoxidans, Burkholderia spp., and Weeksella virosa. TMP-SMZ was active against most species except P. aeruginosa and Pseudomonas fluorescens-putida. A review of laboratory records indicated few changes in susceptibility patterns from 1991 to 1995; the only clear trend was toward increasing P. aeruginosa resistance to all classes of drugs. PMID- 8726012 TI - In vitro synergy testing of macrolide-quinolone combinations against 41 clinical isolates of Legionella. AB - Combination antimicrobial therapy against Legionella species has not been well studied. Several quinolones have activity against Legionella strains, which prompted this in vitro search for a synergistic combination with the macrolides. By a checkerboard assay, erythromycin, clarithromycin, and azithromycin, each in combination with ciprofloxacin and levofloxacin, were tested for synergy against 46 isolates of Legionella. The agar dilution method was employed using buffered charcoal-yeast extract media. A final inoculum of 10(4) CFU per spot was prepared from 24-h growth of each isolate. Plates were incubated at 35 degrees C for 48 h. Synergy, partial synergy, additive effect, or indifference was observed for all combinations of antibiotics tested. There was no antagonism observed. Synergy occurred to a significantly greater extent for the clarithromycin-levofloxacin (P = 0.0001) and azithromycin-levofloxacin (P = 0.003) combinations versus erythromycin-levofloxacin. The azithromycin-ciprofloxacin combination demonstrated significantly greater synergy than did either erythromycin ciprofloxacin (P = 0.003) or clarithromycin-ciprofloxacin (P = 0.001). The newer macrolides clarithromycin and azithromycin may be more active in combination with a fluoroquinolone than is erythromycin. PMID- 8726013 TI - Antimicrobial effect of acidified nitrite on gut pathogens: importance of dietary nitrate in host defense. AB - Dietary intake of nitrate generates salivary nitrite, which is acidified in the stomach, leading to a number of reactive intermediates of nitrogen, among which are the potentially carcinogenic N-nitrosamines. Acidified nitrite, however, also has antimicrobial activity which coincides with the formation of nitric oxide. The present study examines the antimicrobial effect in vitro of acidified nitrite on Salmonella enteritidis, Salmonella typhimurium, Yersinia enterocolitica, Shigella sonnei, and Escherichia coli O157. First-order regression plots showed a linear inverse relationship of log-transformed proton and nitrite concentrations with MICs and MBCs after 30 min, 2 h, and 24 h of exposure (P < 0.001 for all antibacterial activities). Susceptibility to the acidified nitrate solutions ranked as follows: Y. enterocolitica > S. enteritidis > S. typhimurium = Shigella sonnei > E. coli O157 (P < 0.05). Addition of SCN-, but not that of CI-, increased the antibacterial activity (paired t testing, P < 0.001). Generation of salivary nitrite from dietary nitrate may provide significant protection against gut pathogens in humans. PMID- 8726014 TI - Identification, cloning, and expression of the Escherichia coli pyrazinamidase and nicotinamidase gene, pncA. AB - Pyrazinamide (PZA) is one of the three most important drugs for treatment of Mycobacterium tuberculosis infections. The antibacterial activity of PZA requires a bacterial enzyme, pyrazinamidase (PZAase), which hydrolyzes PZA to form pyrazinoic acid and ammonia. Most PZA-resistant clinical M. tuberculosis isolates lack PZAase activity. With the goal of eventually identifying and characterizing the M.tuberculosis PZAase gene, we began with the more tractable organism, Escherichia coli, which also has PZAase activity. We screened a transposon generated E. coli insertion mutant library, using a qualitative PZAase assay. Two PZAase-negative mutants out of 4,000 colonies screened were identified. In each mutant, the transposon interrupted the same 639-bp open reading frame (ORF), ORF1. The expression of ORF1 on a multicopy plasmid complemented a PZAase negative mutant, leading to PZAase activity levels approximately 10-fold greater than those of the wild type. PZA has a structure similar to that of nicotinamide, a pyridine nucleotide cycle intermediate, so we tested our strains for nicotinamidase activity (EC 3.5.1.19) (genetic locus pncA). The construct with multiple plasmid copies of ORF1 had an approximately 10-fold increase in levels of nicotinamidase activity. This overexpressing strain could utilize nicotinamide as a sole nitrogen source, through wild-type E. coli cannot. We conclude that a single E. coli enzyme accounts for both PZAase and nicotinamidase activities and that ORF1 is the E.coli PZAase and nicotinamidase gene, pncA. PMID- 8726015 TI - Antibiotic-impregnated heart valve sewing rings for treatment and prophylaxis of bacterial endocarditis. AB - Prosthetic heart valve sewing rings were impregnated with gentamicin crobefat (EMD 46217), a poorly soluble gentamicin salt, gentamicin sulfate, and clindamycin palmitate to prevent early prosthetic endocarditis. MICs and MBCs of gentamicin and/or clindamycin were tested against several pathogens of early prosthetic endocarditis. The combination of gentamicin and clindamycin was found to be effective against most relevant bacterial pathogens. With an in vitro pharmacokinetic model, the antibacterial activity of gentamicin and clindamycin was tested against Staphylococcus aureus and Escherichia coli. High gentamicin levels over the first 24 h were required for a strong reduction of bacterial counts of both strains. Equal amounts of gentamicin and clindamycin sustained the antibacterial effect and prevented regrowth. The most effective release curves of gentamicin and clindamycin found with an in vitro model were used for monitoring release profiles of these antibiotics from impregnated sewing rings by investigating combinations of gentamicin sulfate, gentamicin crobefat, and clindamycin palmitate. Sewing rings impregnated with 4 mg of gentamicin sulfate, 14 mg of gentamicin crobefat, and 20 mg of clindamycin palmitate gave an initial gentamicin burst and afterwards yielded a lower sustained release of gentamicin and clindamycin palmitate. These in vitro release kinetics were confirmed in vivo by pharmacokinetic analysis after intramuscular implantation of impregnated sewing ring segments. Gentamicin and active clindamycin palmitate metabolites were obtained at the implantation site for at least 2 weeks in concentrations of 3 and 5 micrograms per g of muscle, respectively. The investigated method of impregnation holds promise for revision implants after prosthetic valve endocarditis. It may also serve as a prophylactic tool for routine use against this disease. PMID- 8726016 TI - Indole and (E)-2-hexenal, phytochemical potentiators of polymyxins against Pseudomonas aeruginosa and Escherichia coli. AB - Combinations of polymyxins and phytochemicals were tested for antimicrobial activity against two gram-negative bacteria. Various degrees of potentiation were found against Pseudomonas aeruginosa and Escherichia coli with (E)-2-hexenal and indole. Three-compound combinations were found to further increase the activity of polymyxin B sulfate and colistin methanesulfonate against both bacteria. Combinations with colistin against P. aeruginosa resulted in the highest degree of potentiation, with a 512-fold increase in colistin antimicrobial activity. These results indicate the potential efficacy of phytochemical combinations with antibiotics to enhance total biological activity. PMID- 8726017 TI - In vitro trypanocidal activities of new S-adenosylmethionine decarboxylase inhibitors. AB - A series of novel aromatic derivatives based on the structure of methylglyoxal bis(guanylhydrazone) (MGBG) was examined for in vitro antitrypanosomal activities and cytotoxicities for human cells. One-third of the compounds tested showed trypanocidal activity at concentrations below 0.5 microM after an incubation period of 72 h. Structure-activity analysis revealed that bicyclic compounds with homocyclic rings and unmodified termini were the most active compounds. Results obtained in three laboratories employing different methods and trypanosome populations consistently ranked compound CGP 40215A highest. This compound had a 50% inhibitory concentration of 0.0045 microM for Trypanosoma brucei rhodesiense, was also active against other trypanosome species, including a multidrug resistant Trypanosoma brucei brucei, and was significantly less toxic than other compounds tested for a human adenocarcinoma cell line, with a 50% inhibitory concentration of 1.14 mM. The effect of CGP 40215A was time and dose dependent, and low concentrations of the compound required exposure times of > 2 days to exert trypanocidal activity. Compounds were inactive against Leishmania donovani and Trypanosoma cruzi amastigotes in murine macrophages in vitro. PMID- 8726019 TI - Identification of novel thiocarboxanilide derivatives that suppress a variety of drug-resistant mutant human immunodeficiency virus type 1 strains at a potency similar to that for wild-type virus. AB - A large variety of carboxanilide and thiocarboxanilide derivatives in which the original oxathiin or aliphatic moieties present in the prototype compounds UC84 and UC38 were replaced by an (un) substituted furanyl, thienyl, phenyl, or pyrrole entity have been evaluated for activity against wild-type human immunodeficiency virus type 1 strain IIIB [HIV-1 (IIIB)] and a series of mutant virus strains derived thereof. The mutant viruses contained either the Leu-100- >Ile, Lys-103-->Asn, Val-106-->Ala, Glu-138-->Lys, Tyr-181-->Cys, or Tyr-188- >Leu mutation in their reverse transcriptase. Several 3-(2-methylfuranyl)- and 3 (2-methylthienyl)-thiocarboxanilide ester, (thio)ether, and oxime ether derivatives showed exquisitely potent antiviral activity against wild-type HIV-1 (50% effective concentration, 0.009 to 0.021 microM). The pentenylethers of the 2 methylfuranyl and 2-methylthienyl derivatives (i.e., 313, N-[4-chloro-3-(3-methyl 2-butenyloxy)phenyl]- 2-methyl-3-furancarbothioamide or UC-781, and 314, N-[4 chloro-3-(3-methyl-2-butenyloxy)phenyl] -2-methyl-3-thiophenecarbothioamide or UC 82) proved virtually equally inhibitory for wild-type and the Ile-100, Ala-106, and Lys-138 mutant virus strains (50% effective concentration, 0.015 to 0.021 microM). Their inhibitory effect against the Asn-103 and Cys-181 reverse transcriptase mutant virus strains was decreased only four- to sevenfold compared with wildtype virus. UC-781 and UC-82 should be considered potential candidate drugs for the treatment of HIV-1-infected individuals. PMID- 8726018 TI - In vivo trypanocidal activities of new S-adenosylmethionine decarboxylase inhibitors. AB - A series of novel aromatic derivatives based on the structure of methylglyoxal bis(guanylhydrazone) (MGBG) was examined for trypanocidal activities in human and veterinary trypanosomes of African origin. One agent, CGP 40215A, a bicyclic analog of MGBG which also resembles the diamidines diminazene (Berenil) and pentamidine, was curative of infections by 19 isolates of Trypanosoma brucei subspecies as well as a Trypanosoma congolense isolate. Several of these isolates were resistant to standard trypanocides. Curative doses were < or = 25 mg/kg of body weight/day for 3 days in these acute laboratory model infections. In addition, CGP 40215A also cured a model central nervous system infection in combination with the ornithine decarboxylase inhibitor DL-alpha difluoromethylornithine (DFMO; Ornidyl, eflornithine). Curative combinations were 14 days of oral 2% DFMO (approximately 5 g/kg/day) plus 5, 10, or 25 mg/kg/day for 3 or 7 days given by intraperitoneal injection or with a miniosmotic pump. Combinations were most effective if CGP 40215A was given in the second half or at the end of the DFMO regimen. MGBG has modest activity as an inhibitor of trypanosome S-adenosylmethionine decarboxylase (50% inhibitory concentration [IC50]. 130 microM), while CGP 40215A was a more active inhibitor (IC50, 20 microM). Preincubation of trypanosomes with CGP 40215A for 1 h caused a reduction in spermidine content (36%) and an increase in putrescine content (20%), indicating that one possible mechanism of its action may be inhibition of polyamine biosynthesis. PMID- 8726020 TI - Efficiency of nanoparticles as a carrier system for antiviral agents in human immunodeficiency virus-infected human monocytes/macrophages in vitro. AB - Polyhexylcyanoacrylate nanoparticles loaded with either the human immunodeficiency virus (HIV) protease inhibitor saquinavir (Ro 31-8959) or the nucleoside analog zalcitabine (2',3'-dideoxycytidine) were prepared by emulsion polymerization and tested for antiviral activity in primary human monocytes/macrophages in vitro. Both nanoparticulate formulations led to a dose dependent reduction of HIV type 1 antigen production. While nanoparticle-bound zalcitabine showed no superiority to an aqueous solution of the drug, a significantly higher efficacy was observed with saquinavir-loaded nanoparticles. In acutely infected cells, an aqueous solution of saquinavir showed little antiviral activity at concentrations below 10 nM, whereas the nanoparticulate formulation exhibited a good antiviral effect at a concentration of 1 nM and a still-significant antigen reduction at 0.1 nM (50% inhibitory concentrations = 4.23 nM for the free drug and 0.39 nM for the nanoparticle-bound drug). At a concentration of 100 nM, saquinavir was completely inactive in chronically HIV infected macrophages, but when bound to nanoparticles it caused a 35% decrease in antigen production. Using nanoparticles as a drug carrier system could improve the delivery of antiviral agents to the mononuclear phagocyte system in vivo, overcoming pharmacokinetic problems and enhancing the activities of drugs for the treatment of HIV infection and AIDS. PMID- 8726021 TI - Radioimmunoassay for quantitation of 2',3'-didehydro-3'-deoxythymidine (D4T) in human plasma. AB - 2',3'-Didehydro-3'-deoxythymidine (D4T, or stavudine) has been recently approved for the treatment of AIDS. In the present study, a specific and sensitive radioimmunoassay (RIA) was developed for the quantitation of D4T in human plasma. The RIA is a double-antibody competitive binding assay which uses anti-D4T antiserum raised in rabbits as the primary antibody, a tritiated derivative of D4T as the radioactive tracer, and goat anti-rabbit immunoglobulin G as the secondary antibody. No cross-reaction between D4T and various nucleoside analogs was detected. The limit of quantitation of the method approximated 20 ng/ml. Replicate analyses of quality control samples (40 to 3,500 ng/ml) had satisfactory intra- and interassay precision (coefficient of variation, 1.7 to 16.7%) and accuracy (deviation, -9.5 to +21.0). This newly developed RIA was successfully used in the monitoring of plasma drug levels in healthy volunteers receiving an oral dose of D4T. PMID- 8726022 TI - Bisbenzylisoquinolines as modulators of chloroquine resistance in Plasmodium falciparum and multidrug resistance in tumor cells. AB - Ten naturally occurring bisbenzylisoquinolines (BBIQ) and two dihydro derivatives belonging to five BBIQ subgroups were evaluated in vitro for their ability to inhibit Plasmodium falciparum growth and, in drug combination, to reverse the resistance to chloroquine of strain FcB1. The same alkaloids were also assessed in vitro for their potentiating activity against vinblastine with the multidrug resistant clone CCRF-CEM/VLB, established from lymphoblastic acute leukemia. Three of the BBIQ tested had 50% inhibitory concentrations of less than 1 microM. The most potent antimalarial agent was cocsoline (50% inhibitory concentration, 0.22 microM). Regarding the chloroquine-potentiating effect, fangchinoline exhibited the highest biological activity whereas the remaining compounds displayed either antagonistic or slight synergistic effects. Against the multidrug-resistant cancer cell line, fangchinoline was also by far the most active compound. Although there were clear differences between the activities of tested alkaloids, no relevant structure-activity relationship could be established. Nevertheless, fangchinoline appears to be a new biochemical tool able to help in the comprehension of the mechanism of both chloroquine resistance in P. falciparum and multidrug resistance in tumor cells. PMID- 8726023 TI - Bacteriostatic and bactericidal activities of benzoxazinorifamycin KRM-1648 against Mycobacterium tuberculosis and Mycobacterium avium in human macrophages. AB - Inhibitory and bactericidal activities of KRM-1648 were determined against Mycobacterium tuberculosis and M. avium residing in human monocyte-derived macrophages and extracellular M. tuberculosis and M. avium. MICs and MBCs of KRM 1648 against intracellular and extracellular bacteria were substantially lower than those of rifampin. The MICs and MBCs of either drug against the intracellular bacteria were only twofold lower than or equal to the values found for extracellular bacteria. The prolonged effect of KRM-1648 found in this study is probably associated with high ratios of intracellular accumulation, which were 50- to 100-fold higher than that found for rifampin. Further studies on intracellular distribution of KRM-1648 and on the sites of actual interaction between the drug and bacteria residing in macrophages are necessary, as well as evaluation of combined effects of KRM-1648 with other drugs in long-term macrophage culture experiments. PMID- 8726024 TI - Use of PCR and culture to detect Helicobacter pylori in naturally infected cats following triple antimicrobial therapy. AB - Helicobacter pylori causes gastritis and peptic ulcers and is linked to gastric cancer. Domestic cats from a commercial source were found to be naturally infected with H. pylori, and studies were undertaken to eradicate H. pylori from infected cats by using triple antimicrobial therapy. Eight cats infected with H. pylori were used in the study. Six cats received a 21-day course of oral amoxicillin, metronidazole, and omeprazole, and two cats served as controls. Two weeks and 4 weeks posttreatment (p.t.), all six treated cats were negative at several sites (saliva, gastric juice, and gastric mucosa) for H. pylori by culture. However, as determined by PCR with primers specific for the 26-kDa product, the majority of cats at 2 and 4 weeks p.t. had gastric fluid samples which were positive for H. pylori and three of three cats at 2 weeks p.t. had dental plaque which was positive for H. pylori. At 6 weeks p.t., all six cats had H. pylori-negative cultures for samples from several gastric sites taken at necropsy, and only one cat had H. pylori cultured from gastric juice. PCR analysis revealed that five of six cats had H. pylori DNA amplification products from plaque, saliva, and/or gastric fluid samples. Negative bacterial cultures for cats for which there was demonstrable PCR amplification of H. pylori DNA may reflect the inability of in vitro culture techniques to isolate small numbers of H. pylori organisms, focal colonization at sites not cultured, or a failure of the antibiotics to successfully eradicate H. pylori from extragastric sites which allowed subsequent recolonization of the stomach after cessation of therapy. Alternatively, the treatment strategy may have induced in vivo viable but nonculturable coccoid forms of H. pylori. The H. pylori cat model should allow further studies to test these hypotheses as well as the efficacies of other combined therapeutic regimens. Also, because 100% of these cats were naturally infected with H.pylori, this model should prove useful in exploring mechanisms whereby human populations in underdeveloped countries, which have H. pylori infection rates approaching 100%, have a high rate of recurrence of H. pylori infection after use of prescribed antibiotic therapies that successfully eradicate H. pylori in individuals in developed countries. PMID- 8726026 TI - Effect of exogenous glycine on peptidoglycan composition and resistance in a methicillin-resistant Staphylococcus aureus strain. AB - A highly homogeneously methicillin-resistant Staphylococcus aureus strain was grown in the presence of various concentrations of exogenous glycine. Increasing concentrations of glycine in the medium resulted in a decrease in methicillin resistance and the appearance of a heterogeneous resistance phenotype. Parallel to the gradual changes in resistance was an alteration in the muropeptide composition of peptidoglycan. Increasing concentrations of glycine in the medium resulted in peptidoglycan in which muropeptides with a D-alanyl-D-alanine terminus were replaced with D-alanyl-glycine-terminating muropeptides. The disappearance of D-alanyl-D-alanine-terminating muropeptides in peptidoglycan and the concomitant decrease in resistance indicate a central role for D-alanyl-D alanine-terminating precursors in methicillin resistance. PMID- 8726025 TI - Human serum alpha 1 acid glycoprotein reduces uptake, intracellular concentration, and antiviral activity of A-80987, an inhibitor of the human immunodeficiency virus type 1 protease. AB - The therapeutic utility of a human immunodeficiency virus type 1 (HIV-1) protease inhibitor may depend on its intracellular concentration, which is a property of its uptake, metabolism, and/or efflux. Previous studies in our laboratory indicated that the addition of alpha 1 acid glycoprotein (alpha 1 AGP) to the medium markedly increased the amount of the drug required to limit infection in vitro. In this study, physiologically relevant concentrations of alpha 1 AGP and a radiolabeled inhibitor, A-80987, were used to determine both the uptake and activity of the agent in HIV-1-infected human peripheral blood mononuclear cells and cell lines. Both the uptake and efflux of 14C-labeled A-80987 were rapid (t1/2, < 5 min). Uptake of the drug was linearly dependent on the concentration but insensitive to the metabolic inhibitors KF, sodium cyanide, or CCCP (carbonyl cyanide m-chlorophenyl hydrazone). The amount of A-80987 which entered the cells was inversely proportional to the concentration of alpha 1 AGP (r2, 0.99) and directly proportional to the amount of extracellular non-protein-bound drug (r2, 0.99). Most importantly, the antiviral activity of the drug in HIV-1-infected peripheral blood mononuclear cells and MT-2 cells was directly related to the amount of intracellular A-80987. This study demonstrates that A-80987 binds to alpha 1 AGP, resulting in a free fraction below 10%. Cellular uptake of A-80987 is proportionally decreased in the presence of alpha 1 AGP, which results in less than-expected antiviral activity. Importantly, we demonstrate for the first time that the inhibition of HIV protease is highly correlated with the amount of intracellular inhibitor. PMID- 8726027 TI - Resistance to fluoroquinolones in Escherichia coli isolated from poultry. AB - Quinolone-resistant Escherichia coli strains were isolated from poultry clinical samples in Saudi Arabia. The poultry flocks had been treated with oxolinic acid or flumequine prophylaxis. The measure of the uptake of fluoroquinolones showed that none of the strains had a reduced accumulation of quinolones. The result of complementation with the wild-type E. coli gyrA gene, which restored fluoroquinolone susceptibility, and the isolation of DNA gyrase from six isolates indicated that the resistant strains had an altered DNA gyrase. The minimum effective dose of ciprofloxacin for inhibition of supercoiling catalyzed by the isolated gyrases varied from 0.085 microgram/ml for a susceptible isolate (MIC < 4 micrograms/ml) up to 96 micrograms/ml for the more resistant one (strain 215, MIC > 64 micrograms/ml). For the same two isolates, the minimum effective doses of sparfloxacin varied from 0.17 up to 380 micrograms/ml. The in vitro selection of spontaneous single-step fluoroquinolone-resistant mutants using ciprofloxacin suggested that the more resistant mutants are likely the result of several mutations. These results also show that, as in human medicine, cross-resistance between older quinolones and fluoroquinolones can exist in veterinary isolates and reiterate the need for the prudent use of these drugs. PMID- 8726028 TI - In vivo activities of U-100592 and U-100766, novel oxazolidinone antimicrobial agents, against experimental bacterial infections. AB - The Upjohn oxazolidinones, U-100592 and U-100766, are orally bioavailable synthetic antimicrobial agents with spectra of activity against antibiotic susceptible and -resistant gram-positive pathogens. In several mouse models of methicillin-resistant Staphylococcus aureus infection, U-100592 and U-100766 yielded oral 50% effective doses (ED50) ranging from 1.9 to 8.0 mg/kg of body weight, which compared favorably with vancomycin subcutaneous ED50 values of 1.1 to 4.4 mg/kg. Similarly, both compounds were active versus a Staphylococcus epidermidis experimental systemic infection. U-100592 and U-100766 effectively cured an Enterococcus faecalis systemic infection, with ED50 values of 1.3 and 10.0 mg/kg, and versus a vancomycin-resistant Enterococcus faecium infection in immunocompromised mice, both drugs effected cures at 12.5 and 24.0 mg/kg. Both compounds were exceptionally active in vivo against penicillin- and cephalosporin resistant Streptococcus pneumoniae, with ED50 values ranging from 1.2 to 11.7 mg/kg in systemic infection models. In soft tissue infection models with S. aureus and E. faecalis, both compounds exhibited acceptable curative activities in the range of 11.0 to 39.0 mg/kg. U-100766 was also very active versus the Bacteroides fragilis soft tissue infection model (ED50 = 46.3 mg/kg). In combination-therapy studies, both U-100592 and U-100766 were indifferent or additive in vivo against a monomicrobic S. aureus infection in combination with other antibiotics active against gram-positive bacteria and combined as readily as vancomycin with gentamicin in the treatment of a polymicrobic S. aureus Escherichia coli infection. U-100592 and U-100766 are potent oxazolidinones active against antibiotic-susceptible and -resistant gram-positive pathogens in experimental systemic and soft tissue infections. PMID- 8726029 TI - Pharmacokinetics of lamivudine in human immunodeficiency virus-infected patients with renal dysfunction. AB - The purpose of this study was to determine the safety and pharmacokinetics of lamivudine (3TC), a nucleoside analog that has shown potent in vitro and recent in vivo activity against human immunodeficiency virus. Sixteen human immunodeficiency virus-infected patients, six with normal renal function (creatinine clearance [CLCR], > or = 60 ml/min), four with moderate renal impairment (CLCR, 10 to 40 ml/min), and six with severe renal impairment (CLCR, < 10 ml/min), were enrolled in the study. After an overnight fast, patients were administered 300 mg of 3TC orally. Blood was obtained before 3TC administration and 0.25, 0.5, 0.75, 1, 1.5, 2, 3, 4, 6, 8, 10, 12, 16, 24, 32, 40, and 48 h afterward. Timed urine collections were performed for patients able to produce urine. Serum and urine were assayed for 3TC by reverse-phase high-performance liquid chromatography with UV detection. Pharmacokinetic parameters were calculated by using standard noncompartmental techniques. The peak concentration of 3TC increased with decreasing renal function; geometric means were 2,524, 3,538, and 5,684 ng/ml for patients with normal renal function, moderate renal impairment, and severe renal impairment, respectively. The terminal half-life also increased with decreasing renal function; geometric means were 11.5, 14.1, and 20.7 h for patients with normal renal function, moderate renal impairment, and severe renal impairment, respectively. Both oral and renal clearances were linearly correlated with CLCR. A 300-mg dose of 3TC was well tolerated by all three patient groups. The pharmacokinetics of 3TC is profoundly affected by impaired renal function. Dosage adjustment, by either dose reduction or lengthening of the dosing interval, is warranted. PMID- 8726030 TI - Noncompromised penicillin-resistant pneumococcal pneumonia CBA/J mouse model and comparative efficacies of antibiotics in this model. AB - The present study confirms that CBA/J mice are susceptible to several clinical isolates of Streptococcus pneumoniae, including four of five penicillin susceptible and all five penicillin-resistant strains tested, thus providing the first noncompromised animal model for penicillin-resistant S. pneumoniae pneumonia. In this model, doses of penicillin G of 0.6 mg/kg of body weight given six times at 1-h intervals produced effective pulmonary clearance of a penicillin susceptible strain (penicillin G MIC, 0.015 microgram/ml), while doses of 40 mg/kg given six times at 1-h intervals were required to clear a penicillin resistant strain (penicillin G MIC, 1 microgram/ml). Imipenem (MIC, 0.25 microgram/ml) was the most active antibiotic tested against the penicillin resistant strain, with a calculated dose of 0.42 mg/kg given six times at 1-h intervals, resulting in a 2-log decrease in the number of pulmonary bacteria. Comparable effects were seen with vancomycin (MIC, 0.5 microgram/ml), cefotaxime (MIC, 0.5 microgram/ml), and penicillin G at doses of 3.3, 5.5, and 31.0 mg/kg given six times at 1-h intervals, respectively. The pharmacokinetic profile of vancomycin in infected lungs was superior to those of the other antibiotics, especially in regard to the elimination half-life (215.4 min for vancomycin versus 15.0, 14.5, and 14.5 min for penicillin G, cefotaxime, and imipenem, respectively). Both imipenem and vancomycin allowed 90% survival when 40-mg/kg doses were administered twice a day beginning 5 days after infection. Survival rates with penicillin G (160-mg/kg doses) and cefotaxime (40-mg/kg doses) were 40 and 30%, respectively, while no saline-treated mice survived. The present study shows that the CBA/J mouse pneumonia model may be useful for evaluating antibiotic efficacies against penicillin-resistant pneumococcal pneumonia in immunocompetent individuals. Our data suggest that imipenem and vancomycin may be the most active agents against penicillin-resistant S. pneumoniae pneumonia. PMID- 8726031 TI - Gastrointestinal tract colonization with vancomycin-resistant Enterococcus faecium in an animal model. AB - Vancomycin-resistant enterococci have become important nosocomial pathogens in many institutions. The gastrointestinal tract of susceptible hosts serves as the likely reservoir from which the organism is disseminated. To study factors promoting colonization and the efficacy of decontamination therapy with antimicrobial agents, a model of gastrointestinal colonization with vancomycin resistant Enterococcus faecium was developed in CF1 mice. At baseline, all animals were colonized with non-vancomycin-resistant enterococci (5.0 log10 CFU/g), but vancomycin-resistant organisms were not detectable. Following gastric inoculation with 5 x 10(8) CFU of a clinical isolate of vancomycin-resistant E. faecium, the strain transiently colonized the gastrointestinal tract of 100% of mice but was undetectable by Day 14 (< or = 2.7 log10 mean CFU/g). In animals who received 5 mg of streptomycin per ml or 250 micrograms of vancomycin per ml in drinking water, colonization with the organism occurred at significantly higher bacterial counts than in controls at 7 days following inoculation (9.4 for vancomycin, 9.2 for streptomycin, and 5.1 log10 mean CFU/g for controls; P < 0.05). Fecal concentrations of vancomycin-resistant E. faecium persisted at high counts through Day 22 in mice receiving these antibiotics, but low counts were also still detected in 3 of 10 control animals. In mice with previously established vancomycin-resistant E. faecium colonization, oral administration of ramoplanin, a lipoglycodepsipeptide to which the strain was susceptible, suppressed growth of all enterococci in feces, including the vancomycin-resistant strain after 7 days of therapy (< or = 3.1 and < or = 3.3 log10 mean CFU/g for vancomycin and streptomycin groups, respectively). All mice had a recurrence of colonization with vancomycin-resistant E. faecium after the ramoplanin was discontinued. In summary, this animal model demonstrates the importance of antibiotics in predisposing to gastrointestinal colonization with vancomycin resistant Enterococcus spp. Although treatment with ramoplanin temporarily suppressed the organism, recurrence of colonization due to relapse or reinfection occurred. PMID- 8726032 TI - Oxygen concentration influences proton pump inhibitor activity against Helicobacter pylori in vitro. AB - Omeprazole and lansoprazole are proton pump inhibitors that have shown activity against Helicobacter pylori and other Helicobacter species when tested by agar dilution. Lansoprazole was more active against H. pylori than was omeprazole, and the activity was independent of urease production. Disk susceptibility tests and agar dilution MIC determinations were performed to investigate the effects of incubation under different sets of atmospheric conditions on H. pylori inhibition. Oxygen concentration was found to influence proton pump inhibitor activity in vitro, with higher concentrations leading to greater susceptibility. The method of testing is important in determining the anti-Helicobacter activity of proton pump inhibitors. PMID- 8726033 TI - Vancomycin-gentamicin synergism revisited: effect of gentamicin susceptibility of methicillin-resistant Staphylococcus aureus. AB - Vancomycin monotherapy of deep-seated staphylococcal infection may be associated with poor bacteriological response. We evaluated 24 unique patient isolates of methicillin-resistant Staphylococcus aureus (MRSA) for vancomycin-gentamicin synergism by determining time-kill curves for vancomycin at 10 micrograms/ml and gentamicin at 1 microgram/ml. Nine MRSA strains showed high-level gentamicin resistance (HLGR) (MIC, > 500 micrograms/ml), and 15 did not. Vancomycin gentamicin demonstrated synergism against none of the HLGR strains. For the non HLGR strains, gentamicin agar dilution MICs ranged from 0.5 to > 128 micrograms/ml. Vancomycin-gentamicin demonstrated synergism against six of these strains and indifference against nine of them. There was no relationship between the agar dilution MIC of gentamicin and the occurrence of synergism against non HLGR strains. We conclude that a gentamicin MIC of > 500 micrograms/ml predicts a lack of vancomycin-gentamicin synergism for strains of MRSA. For non-HLGR strains, synergism is not predictable from the gentamicin MIC. PMID- 8726034 TI - Bioluminescence screening in vitro (Bio-Siv) assays for high-volume antimycobacterial drug discovery. AB - Bioluminescence-based assays to indicate antimicrobial susceptibility have been developed and validated for recombinant strains of Mycobacterium tuberculosis, Mycobacterium bovis BCG, Mycobacterium avium, and Mycobacterium intracellulare expressing an integrated eukaryotic luciferase gene. MICs determined with these bioluminescence assays for several antimycobacterial agents, including isoniazid, ethambutol, rifampin, amikacin, streptomycin, ciprofloxacin, and clarithromycin, compared favorably with traditional BACTEC methods and visual estimations of the inhibitory end point. Assay methodology has been optimized for the analysis of large numbers of novel compounds and is simple, inexpensive, and labor efficient. The availability of these four recombinant mycobacteria has permitted a strategy for drug discovery employing the nonpathogenic BCG strain for mass screening purposes with subsequent confirmation of activity against the pathogenic mycobacteria. Furthermore, evidence suggests that the BCG-based screen may allow the direct identification of bactericidal agents. PMID- 8726035 TI - Reporter gene technology to assess activity of antimycobacterial agents in macrophages. AB - Reporter strains of Mycobacterium tuberculosis and Mycobacterium bovis BCG endogenously expressing firefly luciferase were used in bioluminescence assays to evaluate the activities of isoniazid and rifampin against mycobacteria sequestered in human macrophages. This methodology allowed the efficacy of antibiotics against intracellular mycobacteria to be assessed without the labor intensive procedures and protracted incubation requirements associated with conventional CFU determinations. PMID- 8726036 TI - Activity of once-daily cefpodoxime regimens against Haemophilus influenzae and Streptococcus pneumoniae with an in vitro pharmacodynamic chamber model. AB - To characterize the in vitro effectiveness of once-daily dosing with cefpodoxime against Haemophilus influenzae and Streptococcus pneumoniae infections, an in vitro pharmacodynamic chamber model was used to compare the bacterial killing activities of three cefpodoxime regimens: 100 mg twice daily (BID), 200 mg once daily (QD), and 400 mg QD. At the end of 24 h, the regrowth of H. influenzae isolates in the QD regimens was of concern, and the total logarithmic reduction was greatest in the BID regimen (3.1 log). Against S. pneumoniae isolates, the largest reductions in bacterial counts were observed in the 100-mg BID (5.5 log) and 400-mg QD (4.0 log) regimens. These data suggest that 400 mg of cefpodoxime given QD may have a role in the therapy of infections involving S. pneumoniae isolates. PMID- 8726037 TI - Inhibitory effect of erythromycin on interleukin 8 production by 1 alpha,25 dihydroxyvitamin D3-stimulated THP-1 cells. AB - We have recently reported that long-term administration of erythromycin at a low dose reduced the number of neutrophils and concentrations of interleukin 8 (IL-8) in bronchoalveolar lavage fluid in patients with chronic lower respiratory tract disease. To investigate the mechanism of action of erythromycin, we evaluated its effect on IL-8 production in the 1 alpha,25-dihydroxyvitamin D3-stimulated human monocytic cell line THP-1. Erythromycin at a concentration of 10 micrograms/ml significantly reduced IL-8 production by THP-1 cells stimulated with lipopolysaccharide (10 ng/ml) and 1% normal human serum compared with the amount produced by untreated cells (untreated cells, 2,448 pg/ml; erythromycin-treated cells, 872 pg/ml). Our results suggest that erythromycin may impair IL-8 production by alveolar macrophages, ultimately reducing neutrophil accumulation in the airspace. PMID- 8726038 TI - Eucaryotic cells protect Borrelia burgdorferi from the action of penicillin and ceftriaxone but not from the action of doxycycline and erythromycin. AB - Despite appropriate antibiotic treatment, Lyme disease patients may have relapses or may develop chronic manifestations. The intracellular location of Borrelia burgdorferi suggests that antibiotics that penetrate cells will have greater efficiency. Doxycycline or erythromycin was more effective than penicillin or ceftriaxone in killing B. burgdorferi when the organism was grown in the presence of eucaryotic cells. PMID- 8726039 TI - Phosphorylation of the anti-hepatitis B nucleoside analog 1-(2'-deoxy-2'-fluoro-1 beta-D-arabinofuranosyl)-5-iodouracil (FIAU) by human cytosolic and mitochondrial thymidine kinase and implications for cytotoxicity. AB - The capacity of recombinant human cytosolic thymidine kinase (TK1) and bovine mitochondrial thymidine kinase (TK2) to phosphorylate the antiviral analogs 1-(2' deoxy-2'-fluoro-1-beta-D-arabinofuranosyl)-5-iodouracil (FIAU) and 1-(2'-deoxy-2' fluoro-1-beta-D-arabinofuranosyl)-5-methyluracil (FMAU) has been analyzed. The Vmax/Km ratios for FIAU and FMAU with TK2 are about 30% of that for deoxythymidine, while the corresponding values for TK1 are 2 and 5%, respectively. Thus, these two analogs are more efficient substrates for TK2 than for TK1, which may be part of the explanation for the mitochondrial toxicity associated with FIAU during treatment of hepatitis B infection. PMID- 8726040 TI - Distribution of tet(H) among Pasteurella isolates from the United States and Canada. AB - Tetracycline-resistant isolates of Pasteurella multocida and Pasteurella haemolytica obtained from various locations in the United States and Canada were studied to determine the distribution of the tet(H) gene. Of the 31 isolates examined, 25 were found to contain the tet(H) gene. Chromosomal or plasmid DNA obtained from those that did not contain the tet(H) gene did not hybridize with probes specific for classes A through G, though chromosomal DNA from one isolate lacking tet(H) hybridized with a probe specific for class M. The tet(H) gene was found on plasmid as well as on chromosomal DNA, suggesting that it is carried on a transposable element. PMID- 8726041 TI - Sensitization of bacteria to danofloxacin by temperate prophages. AB - Danofloxacin (CP-76,136) is in a class of agents that inhibit DNA gyrase and trigger induction of the SOS response and temperate bacteriophages. Killing studies against the bovine pathogen Pasteurella haemolytica demonstrated that danofloxacin exhibits particularly rapid killing kinetics. Here, lysogenic Escherichia coli bearing lambda is found to be more sensitive to danofloxacin than nonlysogenic E. coli. Danofloxacin exposure also induced a prophage in P. haemolytica. The potency of danofloxacin against lysogens in likely enhanced by this prophage induction. PMID- 8726042 TI - In vitro and in vivo evaluations of LB20304, a new fluoronaphthyridone. AB - In vitro activity of LB20304 against 1,231 clinical isolates was evaluated and compared with those of ciprofloxacin, sparfloxacin, lomefloxacin, and ofloxacin. LB20304 demonstrated the most potent activity against gram-positive bacteria. It was 32- to 64-fold more active than ciprofloxacin against methicillin-susceptible Staphylococcus aureus, methicillin-resistant S. aureus, methicillin-resistant Staphylococcus epidermidis, and Streptococcus pneumoniae (penicillin G resistant). LB20304 was also highly active against most members of the family Enterobacteriaceae. Its activity was more potent than those of sparfloxacin, ofloxacin, and lomefloxacin and comparable to that of ciprofloxacin. The protective activities of LB20304 against systemic infections caused by gram positive bacteria in mice were superior to those of ciprofloxacin and sparfloxacin. Against infections by gram-negative bacteria, LB20304 was slightly less active than ciprofloxacin. PMID- 8726043 TI - Zidovudine does not affect transplacental transfer or systemic clearance of stavudine (2',3'-didehydro-3'-deoxythymidine) in the pigtailed macaque (Macaca nemestrina). AB - Stavudine (22 micrograms/min/kg of body weight) was infused alone (via the femoral vein) or simultaneously with zidovudine (66 micrograms/min/kg) to three near-term pregnant macaques. No significant differences were found between the mean steady-state plasma stavudine concentrations in the dam (Cssd) and fetus (Cssf), the stavudine concentration in the amniotic fluid (Cssa), and the ratios Cssf/Cssd and Cssa/Cssf when stavudine was infused alone or in combination with zidovudine. The data obtained indicate that zidovudine administration does not affect the transfer of stavudine across the placenta in Macaca nemestrina. PMID- 8726044 TI - katG mutations in isoniazid-resistant strains of Mycobacterium tuberculosis are not infrequent. PMID- 8726045 TI - Lack of correlation between in vitro and in vivo studies of combinations of rifampin plus vancomycin or beta-lactam antibiotics against Streptococcus pneumoniae. PMID- 8726046 TI - Marine natural products. Diversity in molecular structure and bioactivity. PMID- 8726047 TI - Plant toxins. The essences of diversity and a challenge to research. PMID- 8726048 TI - Overview of snake venom chemistry. PMID- 8726049 TI - Critical aspects of bacterial protein toxins. PMID- 8726050 TI - Structure and function of cobra neurotoxin. PMID- 8726052 TI - A case study of cardiotoxin III from the Taiwan cobra (Naja naja atra). Solution structure and other physical properties. PMID- 8726051 TI - Structure and function of cobra venom factor, the complement-activating protein in cobra venom. PMID- 8726053 TI - The staphylococcal and streptococcal pyrogenic toxin family. PMID- 8726054 TI - Primary structural motifs of Conus peptides. PMID- 8726055 TI - Hymenoptera venom proteins. PMID- 8726056 TI - Structure and functions of coagulation factor IX/factor X-binding protein isolated from the venom of Trimeresurus flavoviridis. PMID- 8726057 TI - Structure and function relationship of crotoxin, a heterodimeric neurotoxic phospholipase A2 from the venom of a South-American rattlesnake. PMID- 8726058 TI - Atroxase--a fibrinolytic enzyme isolated from the venom of western diamondback rattlesnake. Isolation, characterization and cloning. PMID- 8726059 TI - Isolation of a novel lectin from the globiferous pedicellariae of the sea urchin Toxopneustes pileolus. AB - Sea urchin lectin-I (SUL-I), a 32 kDa lectin was purified from the large globiferous pedicellariae of the sea urchin, Toxopneustes pileolus by using gel permeation chromatography, ion-exchange chromatography and reverse-phase HPLC. SDS-PAGE showed that SUL-I is a monomeric protein with a molecular mass of 32 kDa. Amino acid analysis indicates SUL-I to contain 294 residues. SUL-I was shown to have chemotactic properties for guinea-pig neutrophils at concentrations of 0.625 microgram/ml. These data suggest that a 32 kDa lectin from T. pileolus may be related to defensive role. PMID- 8726060 TI - Indian catfish (Plotosus canius, Hamilton) venom. Occurrence of lethal protein toxin (toxin-PC). PMID- 8726061 TI - Neurotoxin from black widow spider venom. Structure and function. PMID- 8726062 TI - Structural and functional studies of latrodectin from the venom of black widow spider (Latrodectus tredecimguttatus). PMID- 8726063 TI - Effects of toxic shock syndrome toxin-1 and a site-directed mutant, H135A, in mice. PMID- 8726064 TI - The relationship between histidine residues and various biological activities of Clostridium perfringens alpha toxin. PMID- 8726065 TI - Mechanism of action of Clostridium perfringens enterotoxin. PMID- 8726066 TI - Binding proteins on synaptic membranes for certain phospholipases A2 with presynaptic toxicity. PMID- 8726067 TI - Pyrularia thionin. Physical properties, binding to phospholipid bilayers and cellular responses. PMID- 8726068 TI - The chemistry and biological activities of the natural products AAL-toxin and the fumonisins. PMID- 8726069 TI - New aspects of amanitin and phalloidin poisoning. PMID- 8726070 TI - Actions of banana tree extract on smooth and cardiac muscles and in the anesthetized rat. PMID- 8726071 TI - The early expression of myotoxicity and localization of the binding sites of notexin in the soleus muscle of the rat. Notexin and muscle. PMID- 8726072 TI - Fumonisin B1-immunological effects. The influence of FB1 on the early stage of immune response. PMID- 8726073 TI - Biochemical studies on the effect of Plotosus lineatus crude venom (in vivo) and its effect on EAC-cells (in vitro). PMID- 8726074 TI - Interaction of lipopolysaccharide with the antimicrobial peptide "cecropin A". PMID- 8726075 TI - Study on the action mechanism of hemorrhagin I from Agkistrodon acutus venom. PMID- 8726076 TI - K252a and staurosporine microbial alkaloid toxins as prototype of neurotropic drugs. AB - K252 family of alkaloid toxins-kinase inhibitors are the most widely used compounds in biological research on the role of protein kinases in cellular transduction systems, biological functions and pathophysiology of neurological disorders. The wide research interest in these toxins is due to their potency in inhibiting cellular protein kinases. However, lack of kinase specificity is one of their major drawbacks. Synthesis of new K252 derivatives can be expected to open up a new generation of kinase inhibitors. Staurosporine might be considered as a prototype neurotropic drug in view of its ability to induce neurite outgrowth and to increase tau protein levels. Because it mimics some of the neuroprotective effects of NGF and might blocks certain signals required to enhance cellular levels and/or beta amyloid processing, staurosporine might play a beneficial role in the treatment of Alzheimer's disease. The ability of staurosporine to promote neuronal regeneration and brain cholinergic neurons survival has been also demonstrated in animal studies (Nabeshima et al., 1991). The beneficial effects of K252a on the experimental autoimmune encephalomyelitis (EAE) disease mice model and it's ability to supress macrophage activation suggest an important role of protein kinases inhibitors as immunosupressive agents. These results may also point to the potential clinical relevance of K252 microbial toxins as prototypes for the development of new drugs for the management of neuronal diseases. PMID- 8726077 TI - Structure and experimental uses of arthropod venom proteins. AB - In summary, the initial studies conducted thus far into the components of venoms of parasitic wasps and other arthropods have already yielded a number of interesting properties of the proteins therein. These properties have already offered the possibilities of additional principles operating in the evolution of venoms. That so many unexpected rewards have already surfaced with the relatively little experimental digging conducted thus far generates great anticipation that indeed there remains a pharmacological gold mine awaiting to be discovered in components of the other insect venoms as yet unmined by science. PMID- 8726078 TI - Metamorphoses of a conotoxin. PMID- 8726079 TI - Purification and characterization of nerve growth factors (NGFs) from the snake venoms. PMID- 8726080 TI - Snake venoms as probes to study the kinetics of formation and architecture of fibrin network structure. PMID- 8726081 TI - Fibrolase, an active thrombolytic enzyme in arterial and venous thrombosis model systems. PMID- 8726082 TI - Mass spectrometric investigations on proteinaceous toxins and antibodies. PMID- 8726083 TI - Detection of the staphylococcal toxins. PMID- 8726084 TI - Detection and identification of Clostridium botulinum neurotoxins. PMID- 8726085 TI - Detection of botulinum neurotoxins using optical fiber-based biosensor. PMID- 8726086 TI - Comparative studies of antisera against different toxins. PMID- 8726087 TI - New approaches in antivenom therapy. PMID- 8726088 TI - Distribution of domoic acid in seaweeds occurring in Kagoshima, southern Japan. PMID- 8726090 TI - Transcranial Doppler during neurocardiogenic syncope. AB - The purpose of the present study was to investigate changes in cerebral circulation during neurocardiogenic syncope (NCS). Twenty patients with a history of unexplained syncopes were studied over a 45 min period in a tilted position. Heart rate and arterial blood pressure were recorded non-invasively using Finapres. Cerebral blood flow velocity of both middle cerebral arteries was measured with transcranial Doppler (TCD). Ten patients (50%) developed a NCS during the tilt test, with a strong reduction in blood pressure (mean, 48/34 mmHg) and heart rate (mean, 54 beats/min). Simultaneously, diastolic blood flow velocities dropped to values close to zero. However, systolic blood flow velocities did not decrease. In consequence, the pulsatility index (PI) increased considerably from 0.93 to 2.01. The increase in PI suggests that there is a constriction of cerebral resistance vessels during NCS. Despite the drop in blood pressure and the putative increase in cerebrovascular resistance, systolic blood flow velocities remained unchanged in the TCD records. This fact can be explained by a lumen narrowing of the middle cerebral artery at the site of insonation. In conclusion, the typical changes in cerebral blood flow velocity during NCS are probably due to a strong constriction of both the proximal and the peripheral segments of cerebral arteries. It is clear that, in addition to vasodepression and cardiac inhibition, cerebral vasoconstriction is a further mechanism in the pathogenesis of a NCS. PMID- 8726089 TI - QT interval and QT dispersion in multiple system atrophy (Shy-Drager syndrome). AB - To evaluate the influence of autonomic function on the QT interval and QT dispersion, 18 patients (10 males and 8 females; mean age 61 +/- 9 years) with multiple system atrophy (MSA, Shy-Drager syndrome) were studied. Cardiovascular tests were performed to assess the degree of autonomic dysfunction. The QT interval, corrected QT (QTc), QT dispersion (QTd), corrected and adjusted QTd were calculated from a standard 12-lead electrocardiogram. Fifteen healthy subjects matched for sex and age were studied as controls. Nine MSA patients showed severe autonomic dysfunction with orthostatic hypotension. In the remaining patients definite autonomic impairment was found. No statistically significant difference was found in QTd and only a trend towards higher values of maximal QTc was found in patients compared with controls. QTc prolongation, defined as greater than the mean +/- 2 SD of the controls, was detected only in three out of the 18 MSA patients (17%). No correlation was found between the severity of autonomic impairment and repolarization parameters. Our data suggest that chronic autonomic impairment in patients with MSA does not significantly affect ventricular repolarization and ventricular dispersion. PMID- 8726091 TI - Spectral analysis of blood pressure and heart rate, catecholamine and neuropeptide Y plasma levels in a new model of neurogenic orthostatic hypotension in dog. AB - The aim of the study was to compare changes in blood pressure (BP) and heart rate (HR) variability, catecholamine and neuropeptide Y (NPY) plasma levels induced by passive head-up tilt in normal and sino-aortic denervated (SAD) chloralose anaesthetized dogs. In controls, 80 degrees head-up tilt test failed to change BP and increased HR. Plasma noradrenaline and NPY levels (but not adrenaline) significantly rose. In SAD dogs, head-up tilt test induced a marked and reproducible decrease in BP without any change in HR or noradrenaline and NPY plasma levels. In SAD dogs, spectral analysis in supine position was characterized by reduced variability in the high frequency (HF) band of the HR spectrum without changes in low frequency (LF) bands of both HR and systolic blood pressure (SBP). Head-up tilt test increased the LF component of SBP variability and decreased the HF component of HR variability in controls but failed to modify HR and BP variabilities in SAD dogs. In conclusion, sino-aortic denervation in dogs elicits a reproducible postural fall in BP with impaired adaptation of sympathetic nervous system activity. This model may be of value in evaluating the pharmacological effects of drugs for the management of orthostatic hypotension. PMID- 8726092 TI - Cardiovascular adjustments and pain during repeated cold pressor test. AB - The cold pressor test is used in the clinical testing of the autonomic nervous system. However, little is known about changes in the autonomic control of the cardiovascular system during repeated challenge with cold. Heart rate (HR), respiratory sinus arrhythmia (RSA), T-wave amplitude of ECG (TWA), blood pressure, body temperature and perceived pain were recorded in 18 male subjects during three CPTs which consisted of four minutes immersion of the left hand into cold water at 1 degree C. Breathing during CPT was either spontaneous or paced at 0.23 Hz or 0.1 Hz. Pain intensity and HR decreased and TWA increased during the cold immersion and in the resting period preceding cold in the second and third trials. Systolic and pulse blood pressure increased in resting periods in the third trial. RSA increased in the second and third cold challenge during paced breathing at 0.1 Hz only. A decrease in body temperature (0.48 degree C) at the end of the experiment correlated marginally with HR changes. Our study shows that sustained cardiovascular changes are induced by the first challenge with cold, and persist or increase with repeated cold pressor tests. PMID- 8726093 TI - Skin blood flow and autonomic reactivity in human poikilothermia. AB - Autonomic reactivity is pivotal in maintaining a constant body core temperature. Skin vasomotor reflexes and cardiovascular reactivity were investigated in four women (aged 28-37 years) with acquired poikilothermia, during steady-state spontaneous hypothermia (rectal temperature (Tr) = 33.7 +/- 1.0 degrees C [mean +/- SD]) and steady-state normothermia (Tr = 36.7 +/- 0.3 degrees C), as well as in 12 normothermic control subjects. Baseline finger temperature (Tf) during hypothermia was significantly lower than during normothermia (Tf = 32.4 +/- 1.2 compared with 36.2 +/- 0.3 degrees C, respectively), and than in the controls (Tf = 34.8 +/- 0.8 degrees C). No significant differences in baseline skin blood flow and forearm blood flow were found between subjects during hypothermia or normothermia and controls, suggesting a failure of sympathetic drive to counter regulate hypothermia in the subjects. Skin vasoconstrictor responses to the contralateral cooling test and neck cooling test were markedly attenuated in three subjects, and to the finger cooling test in two subjects, during normothermia compared with hypothermia. Blood pressure responses to the Valsalva manoeuvre and head-up tilting were normal in all subjects, whereas the heart rate response to head-up tilting was blunted in three subjects during hypothermia. The responses of blood pressure and forearm blood flow to the cold pressor test in the subjects during both thermal conditions were comparable with the controls. We conclude that in our subjects, without generalized autonomic failure, poikilothermia has to be attributed predominantly to disorders of the central thermoregulatory pathways. Our findings during hypothermia and normothermia indicate that variations in core and skin temperature significantly affect skin vasomotor reactivity. PMID- 8726094 TI - Effects of age on aortic blood velocity at rest and during exercise in patients with coronary artery disease. AB - The effect of age on aortic blood velocity has been studied in 100 patients with angiographically-documented coronary artery disease, 500 of whom were receiving beta-adrenergic blocking agents. Using continuous-wave Doppler ultrasound, the aortic blood velocity signals, both at rest and at maximal-tolerated supine exercise, were obtained. From the Doppler signals the peak velocity (Vp), stroke distance (Sd; the velocity-time integral) and minute distance (Md = Sd x heart rate) were calculated. The measurements were repeated 6 weeks after coronary artery bypass grafting (CABG), performed in 30 patients. No relationship with age (p < 0.01) was found for any of the indices, either at rest or during exercise, except for the resting Md in patients not on beta-blockers, (p < 0.02). No difference in the slope of the relationship with age was found between patients on or not on beta-blockers, except for the resting Md (p < 0.02). Following CABG, a significant age relationship with Vp, Sd and Md was restored, during both resting and exercise, suggesting improvement of systolic left ventricular function following myocardial revascularization. In conclusion, the normal age relationships of the derivatives of aortic blood velocity Doppler ultrasound signals were not seen in patients with coronary artery disease, irrespective of whether they were on or off beta-blockers. The relationship changed following myocardial revascularization, suggesting their dependence on systolic left ventricular function. PMID- 8726095 TI - Possible mechanism of anhidrosis in a symptomatic female carrier of Fabry's disease: an assessment by skin sympathetic nerve activity and sympathetic skin response. AB - Fabry's disease, X-linked alpha-galactosidase deficiency, features a variety of autonomic abnormalities including anhidrosis. In this study, we measured the skin sympathetic nerve activity (SSNA), skin potential and sweat rate in a symptomatic female carrier to investigate the underlying pathophysiology of anhidrosis. The basal activity and responsiveness of SSNA were both fairly well preserved, although slightly reduced compared with the control levels. However, sweating was completely absent, despite the normal skin potential change in response to SSNA bursts. These results suggest that anhidrosis in Fabry's disease is a result of sweat gland dysfunction as well as abnormal SSNA. PMID- 8726096 TI - Effect of meal size on post-prandial blood pressure and on postural hypotension in primary autonomic failure. AB - In chronic autonomic failure, food ingestion causes a profound and rapid fall in supine blood pressure and aggravates postural hypotension. Food volume and caloric load are important determinants of gastric emptying and postprandial splanchnic hyperaemia, which appears to be a major contributor to hypotension. We therefore compared the cardiovascular effects of three large meals with six small meals providing an identical daily caloric intake, in seven subjects with primary autonomic failure. Daytime ambulatory blood pressure (BP) was measured by Spacelabs 90207 every 30 min with additional recordings while lying, sitting and standing, 30 min after each meal. Systolic and diastolic BP were lower in all three positions after large meals; systolic 131 versus 151 mmHg (large versus small), p = 0.005, 109 versus 124 mmHg, 89 versus 103 mmHg and diastolic 76 versus 90 mmHg, p = 0.02, 66 versus 78 mmHg, p = 0.07 and 50 versus 66 mmHg, p = 0.06 for lying, sitting and standing, respectively. Between meals, BP fell to lower levels with large meals, 88 (20) mmHg versus 104 (19) mmHg, p = 0.002 and 48 (13) mmHg versus 63 (13), p = 0.0001 mmHg for systolic and diastolic pressure respectively. Five subjects had more symptoms of postural dizziness after large meals. In primary autonomic failure, smaller and more frequent meals reduce postprandial hypotension and diminish postural symptoms post-meal. This is likely to be a useful non-pharmacological method in the management of postprandial hypotension. PMID- 8726097 TI - Hyperadrenergic state following acute withdrawal from clonidine used at supratherapeutic doses. AB - Abrupt cessation of clonidine treatment precipitates a physiological withdrawal syndrome, thought to be due to a hyperactive state of central autonomic and cognitive adrenergic neuronal systems dependent on presynaptic alpha 2 adrenoceptors and/or imidazoline receptors. We hereby describe a 36-year-old male with history of end-stage renal disease, hypertension and medication non compliance, who presented with severe hypertension and remarkable agitation. His daily clonidine intake was estimated to be 10 mg. The patient had abruptly discontinued his clonidine five days prior to admission. The following indices of adrenergic activity were measured in plasma (normal control values in parentheses): noradrenaline (NA) 8.59 nmol/l (1.32-4.56 nmol/l), adrenaline (Adr) 1.86 nmol/l (0.83-4.20) nmol/l), total 3-methoxy-4-hydroxyphenylglycol (MHPG) 152.8 nmol/l (45.1-111.5 nmol/l), and free MHPG 33.0 nmol/l (12.2-31.4 nmol/l). Plasma clonidine level was 3.53 ng/ml (15.9 nmol/l) with the usual therapeutic level being < 2.0 ng/ml (8.9 nmol/l). Initially, the patient received sedatives and was started on clonidine for the first 24 hours only, after which time period prazosin was started, with good response of his blood pressure and reversal of his mental status changes. At that point, the plasma values of indices of adrenergic activity had decreased compared with their corresponding initial values by the following percentages: NA 60.6%, Adr 22.6%, total MHPG 42.2% and free MHPG 11.5%. Plasma clonidine level had decreased now by 43.6% to an absolute value of 1.99 ng/ml (8.85 nmol/l). We emphasize that physicians should be aware of clonidine's abuse potential and caution should be taken, as well as the appropriate route chosen, when prescribing clonidine in patients who show features of poor compliance to medications and especially in patients with psychoses, suicide potential or personality disorders. PMID- 8726098 TI - Autonomic cardiac abnormalities in alcohol-dependent patients admitted to a psychiatric department. AB - Standardized heart rate analyses were performed in 60 drug-free patients with alcohol dependence, who were admitted consecutively to a psychiatric department, and in 60 healthy normal matched subjects. The procedure included time and frequency derived measurements of heart rate variability (HRV). The investigations were carried out 3 weeks after admission and treatment on a closed ward to avoid autonomic hyperexcitability during withdrawal or relapse. The patients showed a significantly increased heart rate (p < 0.05), a significant reduction in the coefficient of variation while resting (p < 0.01) and a significant decrease of the high frequency power of spectral analysis (p < 0.01) compared with the normal subjects. Cardiovascular autonomic dysfunction was found in 12 of the patients, but in only one of the normal subjects (p < 0.01). The results indicated moderate parasympathetic (vagal) alteration in alcohol dependent patients treated in a psychiatric department. This may have implications for psychotherapeutic or psychopharmacological treatment strategies and prognosis in such patients with alcohol dependence. PMID- 8726099 TI - Subcutaneous botulinum toxin type A inhibits regional sweating: an individual observation. AB - Botulinum toxin inhibits neuromuscular transmission by blocking the exocytosis of acetylcholine. It was tested for a similar effect on cholinergic postganglionic sympathetic neurones at the sudomotor junction. Subcutaneous injections of 0.1 and 1.0 units of type A botulinum toxin into the forearm of a healthy subject abolished local thermoregulatory sweating in cutaneous regions spanning 1.0 and 1.5 cm for nearly 1 year without producing weakness. Botulinum toxin, therefore appears to have potent anhidrotic activity. PMID- 8726100 TI - Consensus statement on the definition of orthostatic hypotension, pure autonomic failure and multiple system atrophy. PMID- 8726101 TI - Control of circadian rhythms by a two-component clock. PMID- 8726102 TI - Localization of cellular storage compartments and sites of constitutive and activity-dependent release of nerve growth factor (NGF) in primary cultures of hippocampal neurons. AB - The characteristics of the constitutive and activity-mediated secretion of NGF from native hippocampal slices are the same as those from hippocampal cultures transfected with an NGF-overexpressing plasmid (Blochl and Thoenen, 1995). In these cultures, the distribution of intracellular NGF immunoreactivity-including the co-localization with endoplasmic reticulum (ER) and Golgi markers-in soma, dendrites, and axons, as visualized by confocal microscopy, is compatible with a localization of NGF in an ER-like compartment. Since the positively charged NGF molecule is bound, at the sites of its release, to the negatively charged neuronal surface, at low salt buffer concentrations, it was possible to attribute the different release mechanisms to specific neuronal surface sites. Constitutive secretion of NGF is confined to the neuronal soma and the very proximal parts of dendrites. In contrast, the activity-dependent secretion, initiated by high potassium or glutamate also occurs all along the neuronal processes, in particular dendrites. This release is independent of extracellular calcium, but depends on calcium released from intracellular calcium stores and is mediated by sodium influx via voltagegated sodium channels and non-NMDA glutamate receptors. The confocal intensity analysis of the NGF surface staining permitted quantitative assessment of the different release mechanisms to different neuronal domains. PMID- 8726103 TI - An in vitro model of the rat dorsal root entry zone reveals developmental changes in the extent of sensory axon growth into the spinal cord. AB - The dorsal root entry zone (DREZ) forms the junction between the dorsal roots of the peripheral nervous system and the spinal cord. In rats older than 1 week, lesioned primary sensory axons regenerate within the dorsal roots but stop at the DREZ, and are thus unable to reconnect with the spinal cord. To analyze the causes of this failure, we have developed a culture model of the interaction of sensory axon growth cones with the intact DREZ, whereby dissociated dorsal root ganglion neurons from rats of various ages are grown on longitudinal cryosections of rat spinal cord, incorporating the DREZ and attached dorsal roots, from neonatal, 1-week-old (P6), or adult animals. Neurites of all ages grew along the roots to the DREZ, where their ability to cross into the spinal cord depended on both their age and that of the spinal cord substrate. Neurites from neonatal neurons failed to cross either the P6 or adult DREZ, but a substantial proportion crossed the immature neonatal DREZ. Early embryonic neurites exhibited substantial crossing on both immature and adult DREZ. These findings strongly suggest that soon after birth, the normal mammalian DREZ acquires growth inhibitory activity that is recognized by the axons of postnatal but not early embryonic sensory neurons. PMID- 8726104 TI - CNTF induces raphe neuronal precursors to switch from a serotonergic to a cholinergic phenotype in vitro. AB - Ciliary neurotrophic factor (CNTF) is a multifunctional cytokine that mediates survival and differentiation of neurons as well as many other cell types. In this study, CNTF and leukemia inhibitory factor (LIF) reduced the apparent number of primary serotonergic neurons in E14 raphe culture by 90% as determined by immunocytochemistry for serotonin (5HT). The reduction in 5HT cell number was not due to neuronal loss as removal of CNTF after 4 days in culture resulted in a partial restitution of the serotonergic phenotype. In the RN46A serotonergic cell line which is induced to become serotonergic by brain-derived neurotrophic factor (BDNF), the addition of CNTF suppressed tryptophan hydroxylase and 5HT synthesis and increased choline acetyl transferase (ChAT) expression by 6-fold and ChAT activity by 20- to 30-fold over 12 days. As with the primary neurons, removal and replacement of CNTF with BDNF after 4 days resulted in a partial restitution of 5HT expression. Moreover, other members of the CNTF-cytokine family that use gp130 and/or LIF receptor beta as their signal transducing receptors-LIF, oncostatin M, interleukin 6, and interleukin 11-had similar effects on increasing ChAT activity and reducing 5HT expression in RN46A cells. Analysis of 5HT levels showed no significant difference in the amount of serotonin between wild-type and CNTFR alpha knockout mice at birth, suggesting that the potential to switch phenotype mediated through CNTFR alpha is a latent property of neuroepithelial precursors in the raphe nucleus. PMID- 8726105 TI - Regulated release and polarized localization of brain-derived neurotrophic factor in hippocampal neurons. AB - The site and regulation of neurotrophic factor release from neurons is poorly understood. We used a combination of model cell lines and primary culture systems to study the polarity of BDNF sorting and the regulation of its release from hippocampal neurons. Transfection and expression of a human BDNF cDNA in a mouse pituitary cell line, AtT20, resulted in the colocalization of BDNF with the secretory granule marker, chromogranin A. Furthermore, stimulation of these cells with 56 mM KCl or with 5 mM 8-bromo-cAMP increased the release of BDNF approximately 10-to 15-fold within 30 min. To study BDNF release from primary cultures of hippocampal neurons, cells were infected with a defective Herpes Simplex Viral (HSV) vector expressing human BDNF. Depolarizing conditions increased the release of BDNF 5-fold from these cells, further verifying that secretion is regulated. Immunocytochemical analysis using highly specific antibodies determined that endogenous BDNF was predominantly localized to the somatodentritic domain of hippocampal neurons. These findings support the view that BDNF functions as a target-derived signal for afferents to hippocampal pyramidal cells and that it may serve as a regulator of hippocampal plasticity. PMID- 8726106 TI - Polyethylenimine-mediated DNA transfection of peripheral and central neurons in primary culture: probing Ca2+ channel structure and function with antisense oligonucleotides. AB - To study neuronal ion channel function with antisense oligonucleotides, a reliable method is needed which allows different neuronal cell types to be transfected without artifactual disruptive effects on their electrical properties. Here we report that use of the recently introduced transfecting agent, polyethylenimine, fulfills this requirement. Four days after transfection, in both central and peripheral neurons, an antisense designed to block the synthesis of the Ca2+ channel beta subunits induced a maximal decrease of the Ca2 current amplitude and modification of their kinetics and voltage-dependence. Controls with scrambled oligonucleotides, as well as Na+ current recordings of antisense transfected neurons, confirm both that the transfecting agent does not modify the electrophysiological properties of the neurons and that the effect of the antisense is sequence specific. PMID- 8726107 TI - Current topics in tardive dyskinesia in Japan. AB - This article reviews current topics in tardive dyskinesia (TD), a movement disorder associated with the prolonged use of neuroleptic agents, especially therapeutic and preventive strategies which have been or are now being studied in Japan. Tardive dyskinesia has become a major problem in the clinical psychiatric field since the early 1970s in Japan, lagging behind Western countries by more than 10 years. The average prevalence rate of TD has been estimated as 7.7% in Japan, while it has been reported in the English literature at around 15 to 20%. Clinical trials of treatments for TD have been or are now being performed in Japan with a number of novel compounds, such as ceruletide, meclofenoxate, and rolipram; however, no effective treatment has yet been established and measures to prevent TD have therefore been emphasized. These include (i) the development of new antipsychotic drugs which are free from TD, (ii) the identification of risk factors from prospective longitudinal studies, and (iii) the investigation of genetic variations that could act as a marker to identify especially vulnerable patients within the whole population of patients who need neuroleptic therapy. PMID- 8726108 TI - Morita therapy: 1919 to 1995. AB - Morita therapy was founded in 1919 by Shoma Morita (1874-1938) and is a systematic psychotherapy based on Eastern psychology. Since its founding 75 years ago, the treatment theory, treatment environment, treatment population and its cultural environment have been examined and modified in some areas. In this review, we first examine Morita theory and discuss the changes that have occurred in subsequent theories and methodologies. Our discussion presents the founding of Morita therapy and the characteristics of Morita theory; the practice of Morita therapy; Morita therapy from the perspectives of transcultural psychiatry and comparative psychotherapy; subsequent developments and modifications of Morita therapy; and the biological studies of Morita therapy. We attempt to clarify the universality and uniqueness of Morita therapy and provide a new framework for understanding Morita therapy. PMID- 8726109 TI - Civilization, culture and neurotic depression: in view of the Freudian theory. AB - Our discussion is based on the psychoanalytic theory of Freud. We, however, discuss a case of neurotic depression which is not considered a subtype in Freud's concept of neurosis. We discuss the difference between culture and civilization, referring to what Freud discussed in Totem and Taboo and Moses and Monotheism. We then discuss two matters specific to Japanese culture; the assimilation of foreign cultures, and the sexual differences found in Japanese culture. On the basis of these discussions, we attempt to link these with the problem of structure of the neurotic depression. We conclude with Confucianism which we can consider as relative to the East-Asian world. PMID- 8726110 TI - Application of the relaxation technique in general hospital psychiatry. AB - The present article demonstrates the clinical application and the efficacy of relaxation techniques in general hospital psychiatry in Japan. During April to December 1993, 20 patients were treated with progressive muscle relaxation and "modified' autogenic training therapies. The targeted symptoms were anxiety attacks (or panic attacks), neurotic insomnia, hyperventilation syndrome, agoraphobia, chronic pain, and distress. Most patients (18/20) improved rapidly with the use of these techniques and their excellence encourages increased use in general hospital psychiatry for the following reasons. First, physically ill patients are likely to show unexpected reactions to psychotropic medications. Second, some patients prefer these treatment methods rather than medications because they feel that they are participating voluntarily in their own treatment and also because they are fearful of being addicted to tranquilizers. Third, these treatments have produced rapid improvement especially for patients with panic attacks and/or hyperventilation. This study strongly suggests that progressive muscle relaxation and "modified' autogenic training are simple and useful methods which can be easily employed in the clinical practice of general hospital psychiatry. PMID- 8726111 TI - A clinical case of neuro-Behcet syndrome without mucocutaneo-ocular symptoms: relationship with brain stem encephalitis. AB - The case of a 50 year old man with personality changes, dementia, and brain stem symptoms is presented. Magnetic resonance imaging (MRI) disclosed high signal areas mainly in the brain stem. Both a positive skin prick test and an HLA-B51 were demonstrated. These clinical findings were suggestive of neuro-Behcet syndrome, although there were no mucocutaneo-ocular symptoms characteristically associated with this disease. The relationship between neuro-Behcet syndrome and brain stem encephalitis, including a discrimination from multiple sclerosis, is discussed. PMID- 8726112 TI - Somatoform pain disorder in a case of Klinefelter's syndrome with multiply operated lower back. AB - We report a case of Klinefelter's syndrome with multiply operated low back (MOB). Psychological and/or psychosocial problems related to MOB have been of recent interest in the field of orthopedic surgery. Based on psychiatric interviews, this case was diagnosed as a somatoform pain disorder of the DSM-III-R somatoform disorders. In addition to psychological problems, the pain was partly explicable by severe osteoporosis, which was prematurely caused by endocrinological disturbances associated with Klinefelter's syndrome. Patients with this syndrome are more likely to develop severe osteoporosis. In the presenile period of Klinefelter's syndrome with severe osteoporosis, liaison psychiatrists may pay attention to somatoform disorders (e.g. somatoform pain disorder and conversion disorder) linked with the MO. PMID- 8726113 TI - Magnetic resonance imaging study of the brain in aged volunteers: T2 high intensity lesions and higher order cortical function. AB - The aims of the present study were to clarify the findings of magnetic resonance imaging (MRI) in the aging brain, and to relate the MRI findings to higher order cortical function. A total of 118 healthy aged volunteers (41 men, 77 women) underwent cranial MRI electroencephalography (EEG), Benton visual retention test (BVRT) and interview. The subjects had no past history or clinical evidence of cerebrovascular disorder, head trauma or dementia and were living at home without any difficulty. The majority of the subjects have participated in this series of studies since 1982. Using a 1.5 T superconductive MR instrument, T1-weighted, proton density and T2-weighted images were obtained. The MRI data were rated visually by regarding 12 items according to fixed criteria. T2 high signal intensity (T2HSI) lesions were found in 69.5% of subjects, the prevalence of which increased with age. T2HSI lesions were most frequently found in the basal ganglia (61.9%), followed by the thalamus (39.0%), parietal lobe (37.0%), temporal lobe (12.7%) and pons (8.5%). Among these lesions, lacunar infarction showed low signal intensity in T1-weighted images and was found in 24.6% of subjects, the prevalence also increasing with age. These findings, including brain atrophy determined according to similar criteria, were correlated closely with the subjects' age. The results of BVRT showed a close relation with T2HSI, suggesting that T2HSI may influence cognitive function. When the subjects were classified according to the presence of T2HSI, lacunar infarction and EEG abnormalities, brain atrophy was significantly milder in a group of subjects with T2HSI(-), lacunar infarction(-) and normal EEG than in the other groups. This suggests that changes seemingly representing physiological aging may be promoted by another pathological which also exerts influences on higher order cerebral function. PMID- 8726114 TI - Reliability of the task-related component (P3b) of P3 event-related potentials. AB - Within session and between session reliabilities of the task-related component (P3b) of the P3 measures (amplitude, area and latency) and their habituation across eight sessions separated by 7-10 days, except for an interval of 1 month between the 6th and 7th sessions, were studied based on the difference waves, which were obtained by subtracting the ignored infrequent event-related potentials (ERP) from the target ERP elicited by a standard auditory oddball paradigm with eyes-open or eyes-closed conditions in 10 normal subjects. The within session reliabilities represented as Pearson correlation coefficients (r) were 0.57-0.66 for the three measures except for those for the latency and amplitude under the eyes-closed condition. The between session reliabilities expressed as intraclass correlation coefficients (R) ranged from 0.54 to 0.60 except for that for latency under eyes-closed conditions. Long-term habituation occurred within the first six sessions for the P3b amplitude and area, and dishabituation took place in the 7th session after an interval of 1 month, whereas no such phenomenon was observed for the P3b latency. Implications of the present results are discussed in terms of the clinical application of the P3 measures. PMID- 8726115 TI - Changes in the ratio of urinary alpha 1-microglobulin to ulinastatin levels in patients with Alzheimer-type dementia and vascular dementia. AB - Relationships between urinary levels of alpha 1-microglobulin (alpha 1M) and ulinastatin (UT) in patients with dementia were investigated. There were no significant differences in alpha 1M and UT levels and alpha 1M: UT ratios among three groups: age-matched control subjects, patients with either Alzheimer-type senile dementia (ATD) or vascular dementia (VD). Although a positive correlation was established between alpha 1M and UT levels in these groups, the regression of the demented patients differed significantly from that of controls (P < 0.05). A tendency towards a negative correlation between alpha 1M: UT ratios and the levels of severity or duration of the disease was displayed in the ATD group, whereas a tendency toward a positive correlation between alpha 1M: UT ratios and the levels of severity was observed in the VD group. These results suggest that changes in the relationships between urinary levels of alpha 1M and UT may provide a useful biochemical index for diagnoses of ATD and VD. PMID- 8726117 TI - Induction of Fos expression following anodal polarization in rat brain. AB - Expression of c-fos immunoreactivity was investigated in rat brain after unilateral application of a weak anodal direct current (anodal polarization) to the sensorimotor cortex of rats. Increases in Fosimmunopositive neurons were observed transiently in the neocortex, cingulate cortex, piriform cortex, and hippocampal formation, which were ipsilateral to the polarization, as a function of the duration and intensity of the current applied. It is likely that anodal polarization enhances the neuronal activities in the cortex dependent on polarization paradigms. PMID- 8726116 TI - Levodopa induces AP-1 and CREB DNA-binding activities in the rat striatum. AB - In order to elucidate the effect of levodopa and bromocriptine on the DNA-binding activities of transcription factors, AP-1 and CREB DNA-binding activities were investigated using gel-shift assay. Intraperitoneal administration of 100 mg/kg levodopa with 50 mg/kg benserazide in rats increased both AP-1 and CREB DNA binding activities in the dorsolateral aspect of the striatum. The major proteins composing the increased AP-1 were JunB and JunD. Bromocriptine at doses of 2.5 and 5.0 mg/kg, however, did not increase these binding activities. Present results suggest that levodopa but not bromocriptine induces these transcription regulating proteins in the striatum with normal dopaminergic functioning. PMID- 8726118 TI - Effects of hCRH on sleep and body temperature rhythms. AB - We investigated the effects of human corticotropin-releasing hormone (hCRH) on the sleep structure and body temperature rhythms of seven healthy young men. Polysomnographic recordings were obtained and body temperatures were monitored continuously for 48 h in each subject following the intravenous administration of saline or of 100 micrograms hCRH. The administration of hCRH was associated with a significant phase-advancement in body temperature rhythm vs the saline control. The administration of hCRH affected non-rapid eye movement (NREM) sleep by reducing significantly slow wave sleep (SWS) and sleep efficiency, whereas the percentage of sleep stages 1 and 2 were increased significantly. These changes in body temperature rhythms and sleep structure induced by hCRH resembled those observed in patients with endogenous depression, except for the findings during rapid eye movement (REM) sleep. We hypothesize that a hypersecretion of hCRH in patients with endogenous depression may help to explain the changes in body temperature rhythms and sleep structure often reported in such patients. PMID- 8726119 TI - Treatment of recurrent hypersomnia with methylcobalamin (vitamin B12): a case report. AB - A 32 year old male patient suffered from recurrent hypersomnia for 12 years was successfully treated with vitamin B12. Episodes of hypersomnia lasting a few days occurred repeatedly a few times each year. Furthermore, the frequency of episodes had increased during the last 2 years. During the administration of vitamin B12, the patient had no episodes of hypersomnia for 6 months. In addition, he did not have any episodes during a follow-up observation period of 17 months after cessation of the treatment. Thus, this case suggests that vitamin B12 may be effective for preventing recurrent hypersomnia. PMID- 8726120 TI - Pharmacokinetics of slow-release preparations of sodium valproate. PMID- 8726121 TI - The present and the future of sleep research in Asia. AB - In recent years, recognition of the importance of sleep, not only as a health problem but also as a social problem, has brought about worldwide growing interest in sleep and sleep research. In Asian countries, the number of sleep researchers is increasing rapidly and as a result the Asian Sleep Research Society (ASRS) was founded in 1994. This paper reviews the main topics of basic and clinical sleep research which have been investigated recently in Japan and in Asian countries. The subjects of sleep research in Asia in the future will be, in principle, similar to those in other countries of the world. However, the author suggests several subjects which are more or less specific to Asia and which should be studied in association, such as epidemiological longitudinal study of sleep disorders in developing countries, characteristics of sleep and dreaming in oriental culture and philosophy, etc. The roles of ASRS in dissemination of information, promotion of training and education, and enlightenment of public and government were also emphasized. PMID- 8726122 TI - Japanese dreams: culture and cosmology. AB - Attitudes to dream evaluation vary depending on culture. Dreams are considered important, real, and public in some cultures, but absurd, irrational and personal in others. Japan has its own history of dreaming, which can be well reconstructed due to rich sources of archeological and documentary material. In this paper dream evolution in Japan is described. Phase 1 is the prehistoric Jomon period, where people believed dreams were part of reality. From Phase 2, the sophisticated philosophies of Confucianism, Buddhism and Taoism were introduced and changed the social and mental system of Japan in phase 3. At phase 4, the amalgamation of local and alien cultures occurred and supernatural beliefs prevailed. In this society dreams played a very important role. Phase 5 is the period when the Samurai class ruled Japan. The pragmatic thinking of the Samurai succeeded in fostering good preconditions for the receipt of scientific Western culture in phase 6. The importance of dreams in Japan evolved in such a way. However, the elements of each phase continued and accumulated similar layers. Thus, a majority of the phases seemed to retain animism from the Jomon period. PMID- 8726123 TI - Sleep disorders in Chinese culture: experiences from a study of insomnia in Taiwan. AB - Traditional Chinese culture-constituted health beliefs continue to influence the Taiwanese people after more than one hundred years of contact with Western medicine. Medicine for sleep disorders, as well as psychiatric medicine, meets some specific difficulties in the professional development. A study of insomnia in Taiwan showed that patients might seek help from a traditional physician and visit a modern hospital at the same stage of medication. General internists and neurologists help to differentiate organic conditions underlying sleep problems but may generalize insomnia to a psychogenic illness. The culture-conditioned attribution of insomnia could also exert certain effects upon pharmacotherapeutic response. PMID- 8726124 TI - The fourth state of consciousness: the Thuriya Avastha. AB - Present day neurophysiology stops with attributing thinking processes as the highest level of function of the brain. It has been common knowledge to oriental thinkers for many centuries, that there are many further states of the human mind, culminating in the state of thoughtless awareness; the fourth state of consciousness. This state must have a physiological basis. The complicated structure of the brain, the extravagant abundance of neural and glial elements in the brain, the infinite possibilities of synaptic junctions and synaptic transmission, and the multitude of neurotransmitters and neuromodulators; all these point to the definite possibility of a much greater level of performance and achievement for the human brain than has been apparent so far. Not only the theories but also the experience of Eastern seers have shown that the brain can transcend the boundaries of logic and reason, and experience states of awareness, commonly unrecognized. In the past few decades, knowledge about the functioning of the human brain has been growing exponentially and scientists of diverse disciplines are concentrating on unraveling its mysteries. It is necessary for scientists to investigate this state with all available tools and find the neurophysiological basis of this state. PMID- 8726125 TI - Is sudden unexplained nocturnal death a breathing disorder? AB - Sudden unexplained nocturnal death syndrome (SUNDS), or 'Lai Tai' in northeastern Thai dialects usually occurs during sleep in healthy young adults, mostly male. The characteristics of Lai Tai are similar to SUNDS which has been reported in several southeast Asian countries and in Japan by the name of 'Bangungut' in Tagalog or 'Pokkuri' in Japanese. The condition has been recognized by the people in the northeastern part of Thailand for many years. Elderly people in the region have described 'Lai Tai' victims as making loud groans and showing signs of difficulty in breathing or labored respiration, who become rigid and die. Data, from relatives or those who had witnessed the episodes of deaths, revealed that the deaths usually occurred in the same manner as described by the elderly. The data also reported that the patients were unresponsive and difficult to arouse. From these reports it is likely that the instability of the physiological systems, especially respiration, in particular during the REM phase, may play some roles in precipitating the sudden death. PMID- 8726126 TI - Sleep in Korea: a developmental perspective. AB - Sleep is an ubiquitous phenomenon. It is a developmental product, being subjected to the vicissitudes of human behavior and culture. The author will attempt to elaborate on sleep, sleep disorders, and sleep medicine in Korea from various developmental perspectives (i.e. personal, national and scientific). Korea is a rapidly developing industrialized nation and is now experiencing immense cultural changes which force individuals to change their behavior and value systems. For example, shift working is becoming increasingly popular and early working hours are being adopted by more companies as a measure to win competitions in the international market. In the clinical setting it is observed very easily that patients develop and maintain disturbed sleep-wake rhythm and its consequences. More obstructive sleep apnea, presumably due to obesity, are observed and studied in sleep clinics. The nuclear family system, an inevitable outcome of industrialization, produces some profound difference in sleeping arrangement such as children's earlier separation from parental or grandparental sleeping environment. The question is how these and other industrialization-related changes may affect the incidence and the manifestation of sleep and sleep disorders. In the background there is emerging interest in healthy sleep and sleep disorders, exemplified by increasing coverage of the topics in the mass media and the publication business. Development of sleep medicine/research per se also involves a developmental perspective. Interests in sleep and sleep disorders began sporadically decades ago in Korea and are now actively being organized mainly by the Korean Association of Sleep Medicine and Psychophysiology as a unified developing force. Understanding of sleep and of sleep disorders is not complete without in-depth understanding of culture, philosophy, and tradition from developmental perspectives. Traditional ideas and wisdom from the past are the unavoidable resources for further understanding sleep and developing sleep researches/medicine in Korea as well as in Asia. PMID- 8726127 TI - Acupuncture treatment for insomnia and acupuncture analgesia. AB - Acupuncture is a simple and useful treatment for insomnia, with a success rate of around 90%. The acupuncture points applied vary depending on the doctor and on the case, but the usual points are Shenmen (HT7) and Anmien (extrapoint). The mechanism of this treatment has not been well elucidated. Acupuncture analgesia may be used as a great indicator, because in both practices needles are inserted in deep tissues as the common stimulation. The possible neuronal integration and transmitter effect in the acupuncture treatment for insomnia are discussed. PMID- 8726128 TI - Reliability and validity of the Japanese version of the Yale-Brown Obsessive Compulsive Scale. AB - The reliability and validity of the Japanese version of the Yale-Brown Obsessive Compulsive Scale (JY-BOCS) were determined by 20 raters for 12 Japanese patients with obsessive compulsive disorder at four institutions. Interrater reliability for the total JY-BOCS score was excellent, and the intraclass correlation coefficient was high (ICC = 0.960). Internal consistency was also excellent (Cronbach's alpha = 0.889). Concurrent and discriminant validity of the JY-BOCS was examined by comparing the scores on the JY-BOCS with those on the Maudsley Obsessional Compulsive Inventory (MOCI) and scales for depression and anxiety. A slight correlation was found between scores on the JY-BOCS and MOCI, but no significant correlations were found between scores on the JY-BOCS and those on scales for depression or anxiety. PMID- 8726129 TI - Is mythical delusion a factor blocking schizophrenic symptoms? AB - It has been stated that mythical delusion can block schizophrenic symptoms. However, there are some schizophrenics in whom mythical delusion does not react against schizophrenic symptoms. Three cases are reported of mythical delusion and what conditions are necessary for the mythical delusion to "block" schizophrenic symptoms are considered. Such "blocking' can be found in the mythical delusion which mediates between schizophrenic symptoms and reality. Conversely, when there is no relationship between mythical delusion and schizophrenic symptoms "blocking' cannot be expected. PMID- 8726130 TI - Positron emission tomography and plasma biochemistry findings in schizophrenic patients before and after electroconvulsive therapy. AB - The clinical effects of electroconvulsive therapy (ECT) on the morbidity of paranoid schizophrenic patients were assessed by positron emission tomography (PET) and plasma biochemistry studies before and after ECT. The present study included five patients whose average age was 41.4 years. The average duration of illness was 23.0 years. To avoid any effect of changes in drugs on PET, no changes were made in the medication of any of the five patients during the study period. ECT improved the clinical symptoms in every patient. Regional cerebral blood flow (rCBF) on PET in both temporal lobes and the left cerebellum was higher in paranoid schizophrenia before ECT than in normal subjects, and rCBF after ECT in both frontal lobes, the right temporal lobe and the right putamen was lower than before ECT as mental symptoms improved. These findings suggest high cerebral blood flow volume in paranoid schizophrenia. Plasma biochemistry studies revealed a lower level of 3-methoxy-4-hydroxyphenylglycol (MHPG) after ECT than before ECT, but a higher level of prolactin existed. PMID- 8726131 TI - Event-related changes of background electroencephalogram during contingent negative variation paradigm. AB - The autoregressive model was applied to the background electroencephalogram during the tone(S1)-tone(S2)-respond contingent negative variation paradigm under eyes-open condition. The EEG data were obtained at the scalp sites of Fz, Cz, and Pz in seven normal males. During the first half of the interstimulus interval (ISI) of 2.5 s, Fz-dominant excitatory process was suggested in concordance with an orienting response to stimulus 1. In the second half of the ISI, Pz-dominant inhibitory process was considered. The stimulus 2 presentation and the required motor response promoted the Pz-dominant process. PMID- 8726132 TI - Prolonged silent periods produced by magnetic cortical stimulation in patients with cerebellar ataxia. AB - Magnetic cortical stimulation can produce silent periods (SP) following excitatory motor responses. The SP in eight patients with cerebellar ataxia was examined. The onset latency of the SP in hand muscles after magnetic cortical stimulation was not different from that in control subjects. The duration of the SP was longer than that in control subjects, but the difference was not significant statistically. The end latency of the SP in patients with cerebellar ataxia was more prolonged than that in control subjects. Therefore, the inhibitory function may be enhanced in patients with cerebellar ataxia. PMID- 8726133 TI - A case of primary hyperparathyroidism that had been treated under a diagnosis of depression for 10 years. AB - A 66 year old man who had been treated under a diagnosis of depression for 10 years was referred to the Kyoto Prefectural University of Medicine, Kyoto because of general fatigue and appetite loss. The patient was diagnosed as having primary hyperparathyroidism (PHPT) based on the increased parathyroid hormone (PTH) and serum calcium levels. Computed tomography revealed solitary adenoma of parathyroid gland. The resection of this solitary adenoma improved the PTH and serum calcium concentrations to normal ranges, which resulted in an improvement in his depressive state. This case suggests that ionic calcium levels contribute to the mental symptoms associated with PHPT. As PHPT is curable, the possibility of PHPT should be taken into account when patients have depressive symptoms. PMID- 8726134 TI - Age-related changes in visual function and visual organs of rats. PMID- 8726135 TI - Morphological changes in the hypothalamic suprachiasmatic nucleus and circadian rhythm of locomotor activity in hereditary microphthalmic rats. AB - Analysis of circadian locomotor activity, Golgi-Cox impregnation, and immunohistochemistry were studied on the hereditary microphthalmic rat which congenitally lacked the optic nerve. These blind rats showed free-running circadian rhythms in their locomotor activities. Both the normal and microphthalmic rats had similar ultradian rhythms in addition to circadian rhythms. The neuronal cell population and volume of the hypothalamic suprachiasmatic nucleus (SCN) of the microphthalmic rats were 66% and 71% of those in normal rats, respectively. The number of SCN neurons containing vasoactive intestinal peptide-like immunoreactive substance was dramatically decreased to 35% of that in normal rats. Golgi-Cox impregnation revealed that three types of neurons in the SCN of the microphthalmic rats were consistently distinguished as observed in normal rats. Although there were no changes in the numbers of primary dendrites of the SCN neurons in the microphthalmic and normal rats, the number of secondary and tertiary dendrites in the SCN of the microphthalmic rats was smaller than that of normal rats. These observations suggest that the retinal input may be important for normal morphological formation of the SCN during development, but not for the generation of circadian rhythms and ultradian rhythms. PMID- 8726136 TI - Distribution of immunoreactive atrial and brain natriuretic peptides in the heart of the chicken, quail, snake and frog. AB - The distribution of atrial natriuretic peptide (ANP)- and brain natriuretic peptide (BNP)-granules was examined immunohistochemically and ultrastructurally in the hearts of the chicken, Japanese quail, Japanese rat snake and bull-frog. Moreover, natriuretic peptide (NP)-granules in the cardiocytes were analyzed by ultrastructural morphometry. Immunohistochemically, ANP-immunoreactivity (IR) was not detected in any cardiocytes, but BNP-IR was detectable in most atrial and ventricular cardiocytes of both chicken and quail. In the snake, ANP-IR was seen in most atrial and ventricular cardiocytes, which showed traces and negative in BNP-IR, respectively. Both ANP- and BNP-IR were detected in the atrial and ventricular cardiocytes in the frog. Ultrastructurally, most of NP-granules were found in the perinuclear region in the chicken, quail and snake atrium, but the frog atrial cardiocytes had granules generally dispersing widely in the cell. By ultrastructural morphometry, the number of granules in the atrial cardiocyte was greatest in the frog, followed by the snake, and chicken or quail, in this order. The diameter of granules in the atrial cardiocyte was largest in the snake and reduced via the frog to the chicken or quail. In the ventricular cardiocytes of all species, the number and size of granules were significantly less than that in the atrial ones. These results indicated that the hearts of the chicken and quail contain only BNP, and that there are two different natriuretic peptides, ANP and BNP, in the snake and frog hearts. PMID- 8726137 TI - Atrial natriuretic peptide (ANP)-granules of auricular cardiocytes in dehydrated and rehydrated mice. AB - Atrial natriuretic peptide (ANP)-granules of auricular cardiocytes were examined by immunohistochemistry, transmission electron microscopy and ultrastructural morphometry in dehydrated and rehydrated mice. In addition, plasma ANP and tissue ANP mRNA levels were determined by radioimmunoassay and polymerase chain reaction, respectively. ANP immunoreactivity and the number of granules in the cardiocytes were increased with time in the dehydration group, while plasma ANP and tissue ANP mRNA levels were decreased on day 3 of dehydration. On day 3 of dehydration, the number of ANP-granules (153.2 +/- 8.3, mean +/- standard error) was significantly (p < 0.05) greater than in the control (125.8 +/- 6.7). IN the rehydration group, the immunoreactivity and number of ANP-granules were less than those in the group on day 3 of dehydration. The plasma ANP level during rehydration for 12 hr was slightly elevated in comparison with the group on day 3 of dehydration. The tissue ANP mRNA level after 12 hr of rehydration was lower than that on day 3 of dehydration. The diameter of ANP-granules was significantly (p < 0.01) smaller in all experimental groups than in the control. These findings suggest that synthesis and secretion of ANP are inhibited and ANP-granules are stored in auricular cardiocytes during dehydration. PMID- 8726138 TI - Comparison of mammary gland growth and some physiological parameters in a high mammary tumor strain of SHN mice and ICR, most popular and 'normal' mice. AB - As a possible step in examining the difference between pathological models and most popular and 'normal' models, the mammary gland and some parameters were compared in SHN mice, a high mammary tumor strain, and ICR mice. SHN was lower than ICR in almost all urinary component levels as indicators of general metabolic activity in both the female and the male, and glucose tolerance in the female. Body weight and several organ weights including liver and kidneys were lower in SHN than in ICR. Normal and preneoplastic mammary gland growth and TGF alpha mRNA expression in the glands of the female were much higher in SHN than in ICR. These findings stress the importance of having a right knowledge of the parameters of 'normal' animals, especially for researchers using pathological models. PMID- 8726139 TI - Branching patterns in coronary artery and ischemic areas induced by coronary arterial occlusion in the CLAWN miniature pig. AB - This study of 28 CLAWN miniature pigs (male 17, female 11, mean weight 29 kg) was undertaken to investigate the coronary arterial branching patterns and the ischemic area induced by surgical occlusion of the left anterior descending artery (LAD) and change in the ischemic area over time. These results were compared with those in dogs, which have frequently been used in myocardial ischemic research. Regarding the coronary arterial branching pattern, there were fewer ventricular branches from the right and left coronary arteries than in dogs. The septal branches arose from only the LAD and the posterior descending artery (PD). The largest septal artery branched from the LAD. There were two types of septal artery branching patterns. In approximately 80% of the CLAWN miniature pigs, the PD arose from the right coronary artery (Right dominance). The peculiarity of the coronary arterial branching pattern in the CLAWN miniature pigs was more similar to human beings than to dogs. The ischemic area induced by occlusion at three-fifths distal section of the LAD was 12.1% to 22.6% (mean 17.1%) of the left ventricle. The ischemic area in all animals that died of global left ventricular malfunction and hemodynamic instability after LAD occlusion was more than 25% of the left ventricle. PMID- 8726140 TI - Histochemical, lectin-histochemical and morphometrical characteristics of intestinal goblet cells of germfree and conventional mice. AB - Histochemical, lectin-histochemical and morphometrical studies were carried out on intestinal goblet cells of 8-week-old germfree (GF) and conventional (CV) mice of the BALB/c strain. Except for the reactivity of cecal goblet cells to Dolichos biflorus agglutinin (DBA) and Ulex europeus-I agglutinin (UEA-I), there was no difference between GF and CV mice in histochemical and lectin-histochemical properties. In the cecal mucosa, DBA stained the goblet cells strongly in CV mice but not in GF mice and UEA-I stained the goblet cells strongly in the lower part of crypts in CV mice but only faintly in GF mice. These findings suggest that terminal residues of cecal goblet cell mucin were different in GF and CV mice. Morphometrically, cecal goblet cells were fewer in number and smaller in size in GF mice than in CV mice. In addition, high iron diamine-alcian blue staining made a very clear border between the cecum and colon, because cecal goblet cells were exclusively positive for sulfomucin and colonic goblet cells were predominantly positive for sialomucin. PMID- 8726141 TI - Effects of phenobarbital on aniline metabolism in primary liver cell culture of rats with ethionine-induced liver disorder. AB - In experiment 1, the amount of aniline (AN) metabolites in the primary cell culture medium of the liver cells obtained from ethionine (ET)-treated rats was compared with that of the control (normal) rats. Although the metabolites detected in both groups were p-aminophenol (p-AP), N-acetyl-p-AP (AAP), acetoanilide (AAN), AAP-glucuronide (AAPG), phenylhydroxylamine sulfate (PHAS) and p-AP-glucuronide (p-APG), the amount of AAP was lower and that of p-APG was markedly higher in the ET-treated rats than in the control rats. In experiment 2, phenobarbital (PB) was orally administered to the ET-treated and control rats at a dose of 100 mg/kg. The time course changes in AN metabolites in the primary cell culture medium of liver cells obtained at 2 or 48 hr after PB treatment were compared with those without PB treatment. In the ET-treated rats, the amount of PHAS was slightly higher at 2 hr after PB treatment, and that of AAP was lower and that of p-APG was higher at 48 hr after PB treatment as compared with those without PB treatment. In the control rats, the amounts of AAP, AAN, p-AP and p APG at 2 hr after PB treatment remained lower than those without PB treatment, and that of AAP was markedly lower and that of p-APG was higher at 48 hr after PB treatment as compared with those without PB treatment. These findings indicated greater detoxication in the primary liver cell culture in the ET-treated rats than in the control rats. Furthermore, detoxication was greater in the primary cell culture of liver cell obtained from the ET-treated rats after PB treatment than from those without PB treatment, because the production of acetylates (AAP) decreased and p-APG increased (induction of conjugated enzyme) in the PB treatment group. PMID- 8726142 TI - Magnetic resonance imaging of the internal structure of the mouse fetus. AB - Magnetic resonance imaging (MRI) has potential as an imaging technique in fetal anatomy. In this article are presented images obtained from mouse fetuses at 16 days of pregnancy by means of the ultraconductive MRI system (JEOL AIM270). When fetuses removed from the uterus were fixed with 10% neutral formalin, various organs including heart, liver, lungs and bones were clearly seen, and a clear outline was obtained of the fetal skin and subcutaneous tissue by varying the setting. The same specimens were able to be used for histological evaluation. This MRI system will allow wider application in fetal anatomy and provide additional information about the developing mouse fetus. PMID- 8726143 TI - The brain histamine (HA) and pituitary luteinizing hormone (LH) levels in female rats anesthetized with ether on proestrus. AB - Levels of brain histamine (HA) and pituitary luteinizing hormone (LH) were determined in female rats anesthetized with ether in the afternoon of proestrus. The rats after 6-hr ether anesthesia had an average HA concentration higher than that of non-anesthetized rats in the hypothalamus, but not in the cortex or diencephalon. Ether-anesthetized rats also showed a higher LH level in the pituitary than that of non-anesthetized rats. These findings agree well with our previous observation of an inhibited ovulation associated with a decrease in serum LH in female rats anesthetized with ether in the afternoon of proestrus. Furthermore, it is suggested that the release of pituitary LH into the circulating blood is regulated by the level of HA in the hypothalamus. PMID- 8726144 TI - Effect of insulin on in vitro development of tetraploid mouse embryos. AB - The effect of insulin on in vitro development of early stage tetraploid mouse embryos was examined. Tetraploid embryos were produced at the 2-cell stage by treatment with cytochalasin B for 12 hr, then they were cultured with M16 medium containing 1 microM insulin. Although insulin had no effect on the embryos until 72 hr after hCG injection, at 96 hr the number of cells of insulin (+) tetraploid blastocysts increased 20.2% more than insulin (-) tetraploid embryos. The increase in the number of cells mainly depended on the inner cell mass (ICM), in which insulin (+) cells increase 76.3% more than insulin (-) cells. These effects of insulin on tetraploid embryos were comparable with those on diploid embryos. The present results suggested that the effect of insulin on the cell proliferation of early stage embryos was not affected by tetraploidy, but the increased number of cells in tetraploid blastocysts was significantly smaller than that in diploid embryos. PMID- 8726145 TI - Hepatitogenicity of three plaque purified variants of hepatotropic mouse hepatitis virus, MHV-2 in athymic nude mice. AB - Hepatitogenicity of 3 plaque purified variants of hepatotropic mouse hepatitis virus, MHV-2 were examined in athymic BALB/c-nu/nu mice up to 9 weeks post infection (9WPI). All of the MHV-2S- and MHV-2M-infected mice died with severe acute hepatitis in 3WPI. On the other hand, MHV-2L-infected mice did not die until 9WPI and showed signs of slow-developing chronic hepatitis with persistent infection under low serum virus neutralizing antibody titers. This suggests that MHV-2L-infected athymic nude mice may be useful as a new model of chronic viral hepatitis. PMID- 8726146 TI - A rapid and easy electrophoretic method for detecting biochemical loci of rat (Rattus norvegices). AB - Methods for electrophoresis for the analysis of biochemical marker genes, which are used widely for genetic monitoring of inbred strains of rats, have been complicated by the variation of the gel and electrode buffer and electrophoretic conditions with the enzyme or the protein to be examined. To simplify the methods, we performed electrophoresis under fixed conditions of 200 V and 30 min using cellulose acetate membrane as the gel and veronal solution as the gel and electrode buffer. Good results were obtained concerning 12 loci, namely, Amy1, Cs1, Es1, Es2, Es3, Es4, Fh1, Gc, Hbb, Ldr1, Mup1, and Svp1. This method was applied to 8 inbred strains of rats and confirmed to be practical. PMID- 8726147 TI - Relationship between arterial and end-tidal carbon dioxide pressure during controlled ventilation in porcine neonates. AB - To investigate the validity of ETCO2 in porcine neonates, which have been frequently used as an experimental model for human neonates, the relationship between arterial (PaCO2) and end-tidal PCO2 (ETCO2) in porcine neonates was examined under different respiratory conditions by regulating both inspiratory flow and positive end expiratory pressure (PEEP). The difference between PaCO2 and ETCO2 widened significantly, according to the significant decrease in tidal volume/body weight ratio (TV/BW) caused by the increase of PEEP. A lower correlation between PaCO2 and ETCO2 was observed in < 6 ml/kg than in > or = 6 ml/kg TV/BW. It therefore seems reasonable to conclude that, in porcine neonates, the valid ETCO2 measurements corresponding to PaCO2 would be obtained at > or = 6 ml/kg TV/BW. PMID- 8726148 TI - Immunohistochemical study on the deposition of apolipoprotein E in cerebral and islet amyloidoses in cynomolgus monkeys (Macaca fascicularis). AB - To investigate the role of apolipoprotein E (apo E) in amyloidoses of cynomolgus monkeys, the localization of apo E in cerebral amyloid, including senile plaques and cerebrovascular amyloid, and in islet amyloid was examined immunohistochemically. Mature types of senile plaques with amyloid deposits and cerebrovascular amyloid showed intense immunoreactivity to both antisera to apo E and amyloid beta protein (A beta). In contrast, diffuse plaques without obvious Congophilic amyloid showed weak immunoreactivity to antiserum to apo E, but intense reactivity to antiserum to A beta. In addition, the number of these apo E positive diffuse plaques was small compared with that of A beta-positive plaques. On the other hand, diabetic islet amyloid that was negative with A beta, reacted intensely with antiserum to apo E. These findings suggest that apo E plays an important role in amyloid fibril formation in several types of amyloidoses. PMID- 8726149 TI - Introduction of retired breeder F344/DuCrj rats for aging research. AB - Male retired breeder F344/DuCrj rats of 17 months of age were purchased in three lots and maintained for aging studies until 25 months of age. These rats were compared with male virgin rats of the same strain for survival percentage, body weight and food consumption. In the retired breeders, decrease in body weight and low food consumption were noted after delivery, and one or two months were required for these parameters to return to the delivery level. After recovery, the body weight and food consumption as well as survival percentage in the retired breeders were similar to those in virgins. From our results, we consider that it takes one to two months to acclimatize aged rats. PMID- 8726150 TI - What is the role of upper airway bacterial cultures in patients with cystic fibrosis? PMID- 8726151 TI - Bronchoalveolar lavage or oropharyngeal cultures to identify lower respiratory pathogens in infants with cystic fibrosis. AB - As collections of lower respiratory tract specimens from young children with cystic fibrosis (CF) are difficult, we determined whether oropharyngeal cultures predicted lower airway pathogens. During 1992-1994, 75 of 90 (83%) infants with CF diagnosed by neonatal screening had 150 simultaneous bronchoalveolar lavage (BAL) and oropharyngeal specimens collected for quantitative bacterial culture at a mean age of 17 months (range, 1-52). Ten children undergoing bronchoscopy for stridor served as controls. Total and differential cell counts and interleukin-8 concentrations were measured in BAL fluid. A subset of bacterial pathogens were typed by pulsed field gel electrophoresis. A non-linear relationship with inflammatory markers supported a diagnosis of lower airway infection when > or = 10(5) colony-forming units/ml were detected. This criterion was met in 47 (31%) BAL cultures from 37 (49%) children. Staphylococcus aureus (19%), Pseudomonas aeruginosa (11%), and Hemophilus influenzae (8%) were the major lower airway pathogens. In oropharyngeal cultures, S. aureus (47%), Escherichia coli (23%), H. influenzae (15%), and P. aeruginosa (13%) predominated. The sensitivity, specificity, and positive and negative predictive values of oropharyngeal cultures for pathogens causing lower respiratory infections were 82%, 83%, 41%, and 97%, respectively. When there was agreement between paired oropharyngeal and BAL cultures, genetic fingerprinting showed some strains of the same organism were unrelated. We conclude that oropharyngeal cultures do not reliably predict the presence of bacterial pathogens in the lower airways of young CF children. PMID- 8726152 TI - Aerosol deposition in infants with cystic fibrosis. AB - Twenty asymptomatic infants with cystic fibrosis (CF) were studied to determine the amount of radiolabeled aerosol [99m technetium diethylenetriamine penta acetic acid (Tc99m DTPA)] deposited in the respiratory system and its distribution. Aerosols were generated by jet nebulization systems that were used in the wards and the laboratory. Subjects were studied in three groups: group A (n = 10) was sedated with chloral hydrate; children inhaled an aerosol of 7.7 microns mass median diameter (MMD); group B (n = 5) was not sedated, using the same nebulization system (same aerosol particle size as group A); and group C (n = 5) was not sedated; these children inhaled an aerosol with an MMD of 3.6 microns. Normal saline plus 4 mCi of Tc99m bound to DTPA was added to each nebulizer. A closed system was used to collect the expired aerosol. Radioactivity in each infant and in the equipment was measured with a gamma camera on completion of nebulization. In groups A and B, the percentages of the total dose deposited in the lung were 0.97 +/- 0.35% and 0.76 +/- 0.36%, respectively. In group C, 2.0 +/- 0.71% was deposited in the lung (P < 0.01). Deposition in the nose, mouth, and pharynx was least in group C (P < 0.01). In groups A and B, the intrathoracic deposition occurred predominantly in the trachea and main bronchi, whereas in group C, significantly more aerosol was deposited in the lung region. There was marked inter-subject variability in the percentage of aerosol deposition within the three groups. There was no correlation between percentage of aerosol deposited in the respiratory system and age, height, or weight. Sedation did not have a significant effect on deposition of aerosol in infants. This study indicates that only a small proportion of nebulized solution is deposited in the lungs of infants and that this proportion is influenced by the particle size of the aerosol. The smaller particle size (3.6 microns MMD) was deposited in the lung better than large particles. PMID- 8726153 TI - Airway responsiveness to hyperosmolar saline challenge in cystic fibrosis: a pilot study. AB - Hyperosmolar aerosols are used to assess airway responsiveness in subjects with asthma. Using a 10% NaCl aerosol, we investigated airway responsiveness in 23 cystic fibrosis (CF) subjects (12 females, 11 males; 19.1 +/- 3.3 years) who had asthma-like symptoms. The pre-challenge predicted forced expiratory volume in 1 second (FEV1) was 74.7 +/- 21.5. The aerosol was generated by a MistO2gen 143A ultrasonic nebulizer and inhaled for 0.5, 1, 2, 4, 8, 8, and 8 minutes or part thereof. Spirometry was performed before and 1 minute after each inhalation period. The challenge was stopped when a > or = 20% fall from the baseline FEV1 was recorded, after the last inhalation period, or when requested by the subject. We recorded different responses to 10% NaCl among subjects. In 7, the FEV1 fell progressively throughout the challenge in a manner similar to asthmatics. By contrast, in 15 subjects the FEV1 was higher at the completion of challenge compared to during challenge, i.e., the fall in FEV1 was transient. In 7 of these subjects, the final FEV1 at the end of the challenge was higher than the pre challenge FEV1. We conclude that inhaled 10% hyperosmolar saline causes either progressive and sustained or transient airway narrowing during challenge in the majority of CF subjects. The cause of the transient airway narrowing requires further investigation. PMID- 8726154 TI - Measurement of lung function in awake 2-4-year-old asthmatic children during methacholine challenge and acute asthma: a comparison of the impulse oscillation technique, the interrupter technique, and transcutaneous measurement of oxygen versus whole-body plethysmography. AB - This study evaluated three techniques for testing of lung function in young awake children. We compared measurements by the forced or impulse oscillation technique (IOS), the interrupter technique (IT), and transcutaneous measurements of oxygen (tcPo2) with concomitant measurements of specific airway resistance (sRaw) during methacholine challenge in 20 stable asthmatic children, 2-4 years old. Measurements were performed with all techniques after each dose of methacholine and after inhalation of a bronchodilator. Measurements were carried out during tidal breathing using a face-mask with a built-in mouthpiece. The ranking of sensitivity was as follows: sRaw > IOS, respiratory reactance at 5 Hz (Xrs5) > tcPo2 > interrupter resistance (Rint) > IOS, respiratory resistance at 5 Hz (Rrs5). The sensitivity of sRaw and Xrs5 was not significantly different, but both were significantly more sensitive than Rint and Rrs5; the sensitivity of tcPo2, Rint, and Rrs5 was not significantly different. Measurements in eight of the subjects performed during an episode of acute asthma yielded comparable results in regard to the sensitivity of the techniques. Measurements improved significantly after bronchodilator administration; however, the response to bronchodilator tended to be less during acute asthma and was best demonstrated by a deterioration of tcPo2. All the evaluated techniques reliably reflect short term changes in respiratory function and can provide clinically useful estimates of airway function. The techniques are non-invasive, are not dependent on the active co-operation or sedation of the subjects, and therefore are well suited for routine use in young children. PMID- 8726155 TI - Efficiency of aerosol medication delivery from a metered dose inhaler versus jet nebulizer in infants with bronchopulmonary dysplasia. AB - The best means for optimal delivery of drugs into lungs of infants with bronchopulmonary dysplasia (BPD) is uncertain. We aimed to measure radio-aerosol deposition of salbutamol by jet nebulizer and metered dose inhalers (MDI) in ventilated and non-ventilated BPD infants. In a randomized, crossover sequence, salbutamol lung deposition was measured using an MDI (2 puffs or 200 micrograms) or sidestream jet nebulizer (5 minutes of nebulization with 100 micrograms/kg) in 10 ventilated (mean birthweight, 1,101 g) and 13 non-ventilated (mean birthweight, 1,093 g) prematurely born infants. Non-ventilated infants inhaled aerosol through a face mask, connected to a nebulizer or an MDI and spacer (Aerochamber). Ventilated infants received aerosol from an MDI + MV15 Aerochamber or a nebulizer inserted in the ventilator circuit. Lung deposition by both methods was low: mean (SEM) from the MDI was 0.67 (0.17)% of the actuated dose, and from the nebulizer it was 1.74 (0.21)% and 0.28 (0.04)% of the nebulized and initial reservoir doses, respectively. Corresponding figures for the ventilated infants were 0.98 (0.19)% from the MDI and 0.95 (0.23)% and 0.22 (0.08)% from the nebulizer. In both groups, and for both methods of delivery, there was marked inter-subject variability in lung deposition and a tendency for the aerosol to be distributed to the central lung regions. PMID- 8726156 TI - Nebulized flunisolide in infants and young children with asthma: a pilot study. AB - The role of nebulized flunisolide solution in controlling recurrent respiratory symptoms was assessed in a double-blind placebo-controlled parallel study on 23 infants and small children (mean age, 14.2 months) with bronchial asthma. Five of the 12 children in the placebo group and 1 of the 11 patients on active treatment had to be withdrawn from the study. Flunisolide significantly improved symptom scores of wheezing and cough. The rescue treatments with salbutamol did not differ between the two groups during the study. Parents considered the active treatment effective in all the patients, while the placebo was considered useful in 4 of 7 children. No side effects were detected with either treatments. This study indicates that nebulized flunisolide may be an effective treatment for infants with recurrent wheezing and cough. PMID- 8726157 TI - Liquid assisted ventilation: an alternative ventilatory strategy for acute meconium aspiration injury. AB - Evidence of surfactant inactivation by meconium has led to the use of exogenous surfactant therapy in the management of meconium aspiration syndrome (MAS). Liquid assisted ventilation has been shown to improve the cardiopulmonary function in lungs with high surface tension. We compared exogenous surfactant therapy with liquid assisted ventilation in the management of experimental acute meconium aspiration injury. Thirty-two newborn lambs were ventilated at peak inspiratory pressures of 13-16 cm H2O, positive end expiratory pressure of 3-4 cm H2O, fractional inspired oxygen concentration (FiO2) of 1.0, and a respiratory frequency range between 30 and 35 breaths/min. Baseline arterial blood gases, pulmonary function, and arterial blood pressure measurements were taken. All lambs were given 2-3 ml/kg of an unfiltered 25% meconium solution. Lambs were then randomized into either gas-ventilated meconium control, or one of three treatment groups: 1) surfactant; 2) partial liquid ventilation (PLV); or 3) total liquid ventilation (TLV) for 4 hours after meconium injury. All treated groups demonstrated a significant increase in arterial oxygenation (P < 0.05); surfactant and PLV-treated lambs demonstrated significantly decreased arterial PCO2 (P < 0.05). Compliance in all groups increased compared with injury values; compliance of the TLV group increased more than in all other treatment groups (P < 0.05). In addition, lung histology of the TLV group demonstrated clear, intact alveolar epithelium and homogeneously expanded alveoli, while no such improvement was evident in the other groups. These data suggest roles for both exogenous surfactant therapy and liquid assisted ventilation techniques in the management of MAS. PMID- 8726158 TI - Inspiratory work of breathing in ventilated preterm infants. AB - In ventilated newborns, part of the inspiratory work of breathing (WOB) may be due to the inspiratory efforts preceding inspiratory ventilator flow. This study was designed to quantify the contribution of these efforts to WOB. WOB was evaluated in six intubated preterm infants ventilated by the Drager Babylog 8000. The ventilatory modes studied were intermittent mandatory ventilation (IMV), continuous positive airway pressure (CPAP), and assist-control ventilation at 10 (ACV10) and 15 (ACV15) cmH2O peak pressure. Mouth flow (V) and esophageal pressure (Pe) were recorded, and WOB was estimate from the area delineated by the esophageal pressure-volume curve, where volume is the time integral of V. Calculation of WOB started either at the onset of the infant's inspiratory flow (WOBi), or at the beginning of the infant's inspiratory muscle efforts, detected on Pe and confirmed on the V tracing (WOBm). WOBm was found to be significantly higher than WOBi under all ventilatory conditions studied. The difference in work of breathing (delta W) between WOBm and WOBi did not depend on the type of ventilatory mode. When delta W was related to WOBm, it amounted to about 30% of WOBm in IMV and CPAP, and 60% in ACV (P < 0.05, ACV15 vs. IMV). These results suggest that, in preterm infants connected to a ventilator, inspiratory efforts preceding flow inspiration might account for a large fraction of the inspiratory work of breathing. PMID- 8726159 TI - Surfactant replacement therapy for adult respiratory distress syndrome in children. AB - Surfactant replacement therapy may have a role in the treatment of ARDS in children. The current studies suggest that rapid instillation of exogenous surfactant is more effective than slow tracheal instillation or aerosolized delivery. Studies suggest that exogenous surfactant given early in the development of ARDS is more effective than therapy provided late in the course of the disease. Natural surfactants appear to be more effective than artificial surfactants due to the presence of SP-B and SP-C, which prevent inhibition of the exogenous surfactant by the protein leakage into the alveolus that is characteristic of ARDS. Exogenous surfactant replacement therapy appears to be safe and well tolerated. A surfactant that can be delivered by aerosol would be useful since this is more easily tolerated by the patients, requires less surfactant, and would be more cost effective when compared with tracheal instillation. Aerosolized surfactant could be given to patients who have not yet required mechanical ventilation, thus potentially preventing the progression of the acute lung injury to respiratory failure. The recent failure of a large multi center trial of aerosolized Exosurf for the treatment of sepsis-related ARDS72 may have been due to the failure of the delivery system as opposed to the surfactant used in the trial; therefore, further research into aerosol delivery systems is needed. There may be different responses to exogenous surfactant therapy by patients with ARDS of different etiologies, such as aspiration pneumonia, sepsis, or trauma. Well-planned placebo-controlled trials will be required to determine these differences. The data supporting the role of surfactant replacement for the treatment of ARDS in children is growing. However, before widespread use of surfactant is considered, a multi-center, placebo controlled trial will be required to establish the safety and efficacy of surfactant replacement in such patients. PMID- 8726160 TI - Lung abscess in a patient with cystic fibrosis: case report and review of the literature. PMID- 8726161 TI - An unusual cause of tachypnea. PMID- 8726162 TI - Early ribavirin treatment did not prevent severe disease in high-risk bronchopulmonary dysplasia patients with respiratory syncytial virus infection. PMID- 8726163 TI - Toxicological aspects of topical silver pharmaceuticals. AB - Silver is generally considered to present a relatively low toxic threat to humans because unintentional exposure to large doses of the noble metal is quite rare. However, as the intentional utilization of silver in pharmaceutical preparations and devices increases, subtle toxic effects of silver may be predictable and expected. The present review examines the scientific literature, primarily covering the past 10 years, dealing with reports describing various types of silver toxicity. These reports consist of both in vitro and in vivo data dealing with immunological, mesenchymal, neural, and parenchymal cell types. Particular emphasis is given to (1) the use of silver in topical antimicrobial preparations as toxicity relates to absorption through dermal wounds into the systemic circulation and possible effects on delayed wound healing, (2) possible local silver toxicity via iontophoretic devices, (3) current theories relating to the toxicological mechanism of action of silver. PMID- 8726164 TI - Occupational hazards for the male reproductive system. AB - The etiology of male infertilities is largely undetermined, and our knowledge of exogenous factors affecting the male reproductive system is still limited. In particular, the role of specific environmental and occupational factors is incompletely elucidated. Various occupational (physical and chemical) agents have been shown to affect male reproductive functions in animals, but large differences in reproductive function and/or xenobiotic handling between species limit extrapolation to humans. When available, human data are often conflicting and, except in a few instances, usually refer to broad and heterogenous occupational categories or to groups of agents (e.g., solvents). It is often difficult to elucidate the role of a single agent because occupational exposure conditions are often complex and various confounding factors related to lifestyle (smoking, alcohol, and diet) or socioeconomic state may also affect sperm quality, fertility, or pregnancy outcomes. The objective of this work is to summarize the main epidemiological and, where relevant, experimental findings pertaining to agents (physical and chemical) encountered in the occupational environment that might affect the male reproductive system (sperm count, motility and morphology, libido, and fertility) and/or related pregnancy outcomes (spontaneous abortion, stillbirth, low birth weight, and birth defects and childhood malignancy in offspring). Some methodological issues related to research on the reproductive effects of toxicants are also discussed briefly. PMID- 8726165 TI - Biotransformation and membrane transport in nephrotoxicity. AB - The kidney is a frequent target organ for toxic effects of xenobiotics. In recent years, the molecular mechanisms responsible for the selective renal toxicity of many nephrotoxic xenobiotics have been elucidated. Accumulation by renal transport mechanisms, and thus aspects of renal physiology, plays an important role in the renal toxicity of some antibiotics, metals, and agents binding to low molecular weight proteins such as alpha(2u)-globulin. The accumulation by active transport of metabolites formed in other organs is involved in the kidney specific toxicity of certain polyhaloalkanes, polyhaloalkenes, hydroquinones, and aminophenols. Other xenobiotics are selectively metabolized to reactive electrophiles by enzymes expressed in the kidney. This review summarizes the present knowledge on the mechanistic basis of target organ selectivity of these compounds. PMID- 8726166 TI - Environmental estrogenic effects of alkylphenol ethoxylates. AB - Alkylphenol ethoxylates (APEs) and related compounds recently have been reported to be estrogenic because it has been demonstrated in laboratory studies that they mimic the effects of estradiol both in vitro and in vivo. Chemicals referred to as "environmental estrogens" are suspected of causing health effects in both humans and wildlife through disruption of the endocrine system. In this review, the occurrence, environmental fate, and biological effects of APEs are presented. To provide understanding of the potential for endocrine disruption due to environmental estrogens, the physiology of estrogens in mammals and fish is also reviewed. The estrogenic potency of other environmental estrogens is compared to the potency of APE degradation products. The reproductive effects of estrogenic compounds are considered when evaluating the potential health effects of APEs. Given the reported environmental concentrations and bioconcentration factors of APE products, the potential for these compounds to produce estrogenic effects in the environment appears low. Although questions concerning the physiological effects of APEs and other environmental estrogens remain unanswered, there are indications that research is in progress that will lead to better understanding of the risks to humans and wildlife. PMID- 8726167 TI - Daily peaks in the incidence of sudden cardiac death and fatal stroke in Niigata Prefecture. AB - To clarify the circadian variation in sudden death (SD) in Japan, where the causes of sudden death differ from those in the USA, we examined all of the death certificates from 1984 to 1986 in Niigata Prefecture, Japan. We defined SD as death which occurred within 1 h from the onset of the underlying cause. A significant circadian variation, with a high incidence between 6 and 8 am and a secondary peak between 6 and 8 pm, was found in the occurrence of sudden cardiac death (SCD, n = 2953). Although the proportion of SCD due to acute myocardial infarction (AMI) was as low as 28% of SCD cases, the circadian variation of SCD was similar to that previously reported in the USA. In SCD due to AMI in males (n = 487), a significant circadian variation with 3 peaks, including a primary peak between 4 and 6 am, was evident. There was also a marked increase in the incidence of fatal stroke between 6 and 8 pm (n = 529). We concluded that 1) a circadian variation with two peak incidences, one between 6 and 8 am, and one between 6 and 8 pm, was characteristic of SCD in general, 2) there was a primary peak between 4 and 6 am for SCD due to AMI in males, and 3) there was a peak between 6 and 8 pm in the incidence of fatal stroke for both men and women. PMID- 8726168 TI - Diagnostic significance of thrombin-antithrombin III complex (TAT) and D-dimer in patients with deep venous thrombosis. AB - Thrombin-antithrombin III (TAT) and D-dimer were measured in 50 patients suspected of deep venous thrombosis (DVT) to assess the usefulness of these indicators in the diagnosis of DVT. DVT was diagnosed by ultrasonography (compression method and Doppler imaging). In patients who were negative for DVT (Group A), TAT was 3.8 +/- 2.36 micrograms/L (mean +/- SD) and D-dimer was 0.7 +/ 0.69 micrograms/ml, whereas in patients diagnosed with DVT (Group B), TAT was 20.4 +/- 19.10 micrograms/L (p < 0.001) and D-dimer was 9.0 +/- 9.21 micrograms/ml (p < 0.001). Thus, Group B had significantly higher levels of both markers. Moreover, 19 of the 23 cases in Group B had acute DVT, with symptoms appearing within 2 weeks of onset. When the cutoff for a positive diagnosis of DVT was set at TAT of 7.0 micrograms/L or more and D-dimer of 3.0 micrograms/ml or more, sensitivity was 84%, specificity was 96%, and accuracy was 90%. Based on these results, we concluded that TAT and D-dimer are extremely useful in screening for acute DVT. PMID- 8726169 TI - Effect of cilostazol, a novel anti-platelet drug, on restenosis after percutaneous transluminal coronary angioplasty. AB - The possible preventive effect of cilostazol, a novel anti-platelet drug, on restenosis after successful percutaneous transluminal coronary angioplasty (PTCA) was examined. One hundred and two consecutive patients, who underwent successful PTCA, were followed for 3 to 6 months. To prevent restenosis, 46 patients (60 PTCA sites) were treated with cilostazol alone (200 mg/day) (cilostazol group) and the remaining 56 (61 PTCA sites) were treated with other anti-platelet drugs and/or warfarin potassium (control group). Restenosis was defined as a more than 50% loss of the initial gain of the coronary diameter achieved by PTCA. Cilostazol did not significantly reduce the patient or lesion restenosis rate; the patient restenosis rate was 32% in the control group and 22% in the cilostazol group (P = 0.24), and the lesion restenosis rate was 30% in the control group and 23% in the cilostazol group (P = 0.44). However, the lesion non progression rate, which was defined as the incidence of lesions with either no change or regression of coronary stenosis at the PTCA site, was significantly greater with cilostazol (37%) than in the control group (16%) (p < 0.05). Although cilostazol failed to show a significant reduction in restenosis after PTCA, the present results suggest that a further trial with a larger number of patients is needed to confirm its usefulness. PMID- 8726170 TI - Stroke volume generation of the left ventricle and its relation to chamber shape in normal subjects and patients with mitral or aortic regurgitation. AB - The total stroke volume of the left ventricle (LV) is equal to the sum of the regional cavity shrinkage. Since nonuniformity of regional wall motion in LV has been well documented even in normal subjects, the extent of the contribution of each region to total stroke volume cannot be easily determined. To assess the left ventricular regional contributions to total stroke volume under normal conditions and in compensated chronic mitral or aortic regurgitation, LV cineangiograms were analyzed in 14 normal subjects (N), 8 patients with mitral regurgitation (MR) and 10 patients with aortic regurgitation (AR). We assumed that the LV cavity could be viewed as a stack of 30 half-cylindrical discs, 15 in the anterior and 15 in the inferior wall regions. LV chamber shape was more spherical in MR than in N, but was more conical in AR. Percent regional hemichordal shortening was significantly decreased in the anterobasal and anteroapical walls in AR, but was similar between N and MR. The regional contribution to total stroke volume showed a significant quadratic correlation with the end-diastolic regional shape index (N, r = 0.87; MR, r = 0.79; AR, r = 0.90), which was defined as the regional hemiaxial length divided by the LV long axis length, but was not correlated with percent regional hemichordal shortening. Therefore, stroke volume is generated mainly in the mid-ventricular portion in N and MR, but in the basal portion in AR due to the characteristic change in cavity shape. PMID- 8726171 TI - Criteria for evaluating abnormal signal-averaged electrocardiogram in children classified by age, body surface area and height. AB - We evaluated criteria for the diagnosis of ventricular abnormal signal-averaged electrocardiogram (SAE) in 205 healthy children classified by age, body surface area and height. The children were classified by age into 4 groups: under 3 months old, 3 months old or more but less than 1 year old, 1 year old or more but less than 12 year old and 12 year old or more. They were also divided by body surface area into 4 groups: under 0.3 m2, 0.3 m2 or more but less than 0.5 m2, 0.5 m2 or more but less than 1.2 m2, and 1.2 m2 or more. In terms of height, they were classified into 4 groups: under 60 cm, 60 cm or more but less than 80 cm, 80 cm or more but less than 140 cm, and 140 cm or more. The boundary points in these 3 classifications were statistically consistent. Criteria for abnormal SAE in children classified according to age were filtered QRS duration (f-QRSd) > 95 msec, root mean square (RMS) < 30 mu v and duration of low amplitude signal (LAS) > 25 msec in those under 3 months old; f-QRSd > 110 msec, RMS < 25 mu v, LAS > 30 msec in those 3 months or more but less than 1 year old; f-QRSd > 115 msec, RMS < 20 mu v, LAS > 30 msec in those 1 year old or more but less than 12 year old; and f-QRSd > 125 msec, RMS < 20 mu v, LAS > 30 msec in those 12 year old or more. The criteria for evaluating abnormal SAE were similar regardless of whether children were classified by age, body surface area or height. Using these values as criteria, the 205 children were evaluated for the presence of abnormal SAE. All of the children were negative for abnormal SAE using these criteria according to age, body surface area or height. In evaluating abnormal SAE in children, it is useful to classify children by age, body surface area and height and to identify the criteria in each group. In this study, the criteria based on height and age were particularly useful, since these are more convenient than body surface area. PMID- 8726172 TI - The role of extracellular cations in the development of myogenic contraction in isolated rat small arteries. AB - The purpose of this study was to determine the roles of extracellular cations (Na+, Ca2+ and K+), membrane K+ channels and Na+/K+ ATPase in the development of myogenic contraction (transmural pressure-induced contraction) in isolated rat skeletal muscle and mesenteric small arteries. The vessels were pressurized under no-flow conditions in a tissue bath. Lumen diameter was measured with a videomicroscopic system. Myogenic contraction was evoked by increasing the lumen pressure from 40 to 100 mmHg. The vessels demonstrated myogenic contraction in low-Na+ (Na+ 1.18 mmol/L) physiological salt solution (PSS), and this was abolished by removing Ca2+ or by applying nifedipine or nisoldipine (10 mumol/L). Neither tetraethylammonium (TEA, 1 mmol/L), Ba2+ (10 mumol/L) nor glibenclamide (1 mumol/L) affected the magnitude of the myogenic contraction. K(+)-free PSS and ouabain (0.1 mmol/L) partially depressed myogenic contraction. In conclusion, myogenic contraction was triggered by a cellular process that requires extracellular Ca2+, but not Na+ or K+. This triggering process is not affected by TEA, Ba2+ or glibenclamide. PMID- 8726173 TI - Reduced vasodilator response of the right coronary artery to myocardial ischemia in the hypertrophied right ventricle. AB - The reduced reactive hyperemic response of the right coronary artery (RCA) to brief coronary occlusion was assessed in dogs with pressure-induced right ventricular hypertrophy (RVH). Right coronary reactive hyperemia was observed in normal dogs and in dogs with pressure-induced RVH. RVH was induced by chronic pulmonary artery banding in eight 3- to 6-month-old dogs, and reactive hyperemia responses to coronary occlusion lasting for 5, 10, 15, 20, 30, 45, and 60 sec were compared to those in normal dogs. In dogs with RVH, the peak reactive flow rate and excess blood flow debt repayment after the release of 5- to 60 sec RCA occlusion were markedly attenuated. The calculated minimum coronary resistance was higher in RVH dogs than in normal dogs (p < 0.02). The occlusion time that produced one-half of the maximum %PRHc, T1/2, was significantly (p < 0.01) shorter in RVH dogs than in normal dogs, where %PRHc = (peak reactive flow rate baseline flow rate)/(baseline flow rate). T1/2 in RVH dogs varied inversely with right ventricular systolic pressure. Therefore, blood flow in the RCA in RVH is characterized by an attenuated flow response to acute myocardial ischemia, suggesting inadequate development of the coronary vasculature supplying the hypertrophied ventricle. PMID- 8726174 TI - Abnormal Q wave, ST-segment elevation, T-wave inversion, and widespread focal myocytolysis associated with subarachnoid hemorrhage. AB - A 74-year-old Japanese woman with subarachnoid hemorrhage was admitted to our hospital. During her hospitalization, serial electrocardiograms showed the combination of abnormal Q waves, ST-segment elevation, and T-wave inversion, which strongly suggested acute myocardial infarction. However, postmortem examination revealed widespread focal myocytolysis of the myocardium which was unrelated to vascular distribution. PMID- 8726175 TI - Reconstruction of the mitral annulus with porcine pericardium--report of a case with mitral annular disruption due to staphylococcal endocarditis. AB - A 60-year-old man was admitted to our hospital for investigation of dyspnea and disorientation with right hemiplegia. Echocardiography showed thickened mitral valve leaflets with vegetations and severe mitral regurgitation. Blood cultures grew Staphylococcus aureus. During the operation, perforation and destruction of the mitral valve leaflets and vegetations were confirmed. Debridement of the infected tissues resulted in segmental disruption of the posterior mitral fibrous annulus. Reconstruction of the mitral annulus with porcine pericardium treated with glutaraldehyde and mitral valve replacement were successful. The patient's postoperative course was complicated with metastatic cerebral and splenic abscesses. After splenectomy on the 8th postoperative day, he gradually recovered without major neurologic sequelae. We believe that reconstruction of the mitral valve annulus with pericardium, especially autologous pericardium, is reliable and useful for the treatment of patients with disruption of the mitral valve annulus. PMID- 8726176 TI - Role of axillary dissection in breast cancer management. PMID- 8726177 TI - Clinical behavior of untreated axillary nodes after local treatment for primary breast cancer. AB - BACKGROUND: The purpose of this study was to examine the rate of axillary failure in patients with primary breast cancer treated without axillary dissection or radiation and to determine what factors may be associated with axillary failure. METHODS: We studied 112 patients with invasive breast cancer treated for primary disease with breast-conserving surgery without axillary dissection or radiation to the breast or axilla, accrued between 1977 and 1986. Data for these patients were prospectively gathered for a research database and reviewed retrospectively to determine axillary failure. The effects of age, tumor size, estrogen receptor (ER) status, progesterone receptor (PgR) status, histologic grade, nuclear grade, and tumor emboli on time to axillary failure were examined. RESULTS: The median follow-up was 9.6 years. There were 26 axillary recurrences, resulting in a 10 year actuarial nodal control rate of 72%. Patients with nodal failure proceeded to axillary dissection with minimal morbidity. In both univariate and multivariate analyses, only tumor size was significantly associated with axillary failure (p = 0.04 and p = 0.06, respectively). CONCLUSIONS: This study demonstrates a significant effect of tumor size on axillary failure and a reasonable rate of local control in small tumors. Further research should examine the utility of axillary dissection in women with small breast cancers. PMID- 8726178 TI - Molecular diagnosis of exocrine pancreatic cancer using a percutaneous technique. AB - BACKGROUND: The K-ras oncogene is activated by point mutations at codon 12 in most patients with exocrine pancreatic cancer. Mutant-enriched polymerase chain reaction (PCR) amplification can enhance the detection of mutated K-ras. This technique was applied to patients undergoing percutaneous fine-needle aspiration (FNA) biopsy of suspect pancreatic lesions. METHODS: Twenty-five patients underwent percutaneous FNA of the pancreas for cytologic and molecular analysis. After preparing cytologic smears, the 22-gauge needle and syringe used for FNA were rinsed in RPMI-1640. The specimen was centrifuged, and DNA was extracted from the supernatant and subjected to mutant-enriched PCR using appropriate mismatched primers that introduce a BstNI restriction endonuclease clevage site at codon 12 of wild-type, but not mutant, K-ras. After digestion with BstNI, the DNA was reamplified. To increase assay sensitivity, the final five PCR cycles were completed incorporating 5 microCi of (alpha-32P)dCTP. The DNA was then redigested and subjected to gel electrophoresis and autoradiography. RESULTS: The median amount of DNA retrieved per specimen was 3.33 micrograms. Mutant K-ras was detected as a band of 143 bps; residual wild-type DNA was seen as a 114-bp fragment. Twenty-one of 25 specimens demonstrated mutated K-ras DNA. Two patients with nondiagnostic cytology results had mutated K-ras DNA; adenocarcinoma of pancreatic origin was confirmed in both cases after pancreatectomy. CONCLUSION: The molecular diagnosis of pancreatic cancer through identifications of mutations in K-ras can be readily performed on specimens obtained by percutaneous FNA. As aggressive multimodality management of this disease becomes more common, pretreatment analysis of molecular determinants may have greater clinical significance. PMID- 8726179 TI - Immunological memory induced by genetically transduced tumor cells. AB - BACKGROUND: Recent studies have demonstrated the usefulness of gene-modified tumor cells for immunotherapy. Using the tumorigenic murine fibrosarcoma, MCA 106, we investigated the effects of localized interferon-gamma (IFNg) secretion on tumorigenicity and on long-term memory. METHODS: The murine IFNg (MuIFNg) gene was introduced into tumor cells. High and low IFNg-secreting clones were isolated. C57BL/6 mice were injected subcutaneously (s.c.) with either parental (P), high or low IFNg-secreting (H- or L-IFNg) cells, and tumor growth was assessed weekly. Spleens were harvested on different days postinjection (p.i.) to assess in vitro cytolytic activity. In parallel, tissues from injection sites were stained with macrophage-, CD4-, and CD8-detecting antibodies. Mice were injected s.c. with H-IFNg MCA106 tumor. After 150 days the animals were rechallenged s.c. with MCA106P in one leg and with irrelevant syngeneic tumor in the other. RESULTS: Both P- and L-IFNg cells had similar growth, whereas the H IFNg cells never grew. Only splenocytes from the H-IFNg animals showed in vitro CTL activity persisting until day 30 p.i. Histological data revealed a macrophage and CD4+ infiltrate much earlier in the H-IFNg group compared with the P group. Only the irrelevant, syngeneic tumor grew in animals previously injected with H IFNg cells, whereas both P and irrelevant syngeneic tumors grew in controls. CONCLUSIONS: Transduction of MCA106 cells with the MuIFNg gene diminished in vivo tumorigenicity in proportion to the amount of IFNg secreted. Immunization with H IFNg cells elicited a host response characterized by macrophages and CD4+ cells. Long-term tumor-specific memory was seen after immunization with H-IFNg cells. PMID- 8726180 TI - Local recurrence and survival in soft-tissue sarcomas. AB - BACKGROUND: There is a continuous interest in the literature concerning the management and survival after treatment of local recurrence in sarcomas because it is one of the most common types of recurrence. DESIGN: We retrospectively reviewed 93 patients treated for local recurrence from soft-tissue sarcoma. METHODS: We evaluated prognostic parameters (grade, tumor size, location) and the effect of treatment on survival. RESULTS: Resection of all the gross tumor at first visit to our Institute for local recurrence was accomplished in 88 patients (95%). Of the 59 patients with extremity tumors, six (10%) required an amputation. At a mean follow-up of 66 months, further local recurrence was noted in 27%. The estimated 5-year survival rate was 100% for patients with grade I tumors (n = 16), 77% for grade II (n = 31), and 45% for grade III tumors (n = 46) (p = 0.0002). This value was 78% for tumors < or = 5 cm and 57% for those > 5 cm (p = 0.03). CONCLUSIONS: Local recurrence is resectable and limb preservation is possible in the majority of patients. The overall 5-year survival rate was 65%. Survival after treatment of local recurrence is determined mainly by the grade and secondarily by the size of the tumor as for primary sarcomas. PMID- 8726181 TI - Chest wall invasive non-small cell lung cancer: patterns of failure and implications for a revised staging system. AB - BACKGROUND: To assess outcomes and patterns of failure for chest wall invasive non-small cell lung cancer (T3 or IIIA NSCLC), data were acquired prospectively on 47 consecutive patients at a single institution over 6 years. METHODS: Preresectional stagings included bone scan, head and chest/abdominal computed tomography, and mediastinoscopy. There were 25 superior sulcus tumors (radiation and/or chemotherapy followed by resection) and 22 other chest wall invasive NSCLCs (resection alone). RESULTS: There were no perioperative deaths. Seventeen patients (36%) had an operative complication (median length of stay increased from 7 to 12 days; p < 0.05). A complete pathologic resection was achieved for 44 of 47 patients (94%). The median survival was 38 months (actuarial 2- and 5-year survival rates of 62% and 50%, respectively). Median lengths of survival for superior sulcus and other chest wall tumors were 36 and > 60 months, respectively. Significant univariate predictors of decreased overall and cancer free survival were poor performance status, positive margins, and positive lymph nodes. Recurrence was observed in 22 of 47 patients (46%) at a median of 8 months (range 2-24); patterns of failure were in the ipsilateral chest (n = 2; 4%) and at a distant site (n = 15; 32%) or both (n = 5; 11%). CONCLUSIONS: The operative risk for chest wall invasive NSCLC is acceptable, even after neoadjuvant therapy, allowing for a 94% complete resection rate. The survival of this subset of stage IIIA patients may warrant a reappraisal of the international staging system. PMID- 8726182 TI - Suppository delivery of 5-fluorouracil in rectal cancer. AB - BACKGROUND: Preoperative radiotherapy with concomitant intravenous 5-fluorouracil (5-FU-i.v.) is effective in shrinking locally advanced rectal cancers and facilitating subsequent surgery. Topical 5-FU application may enhance its radiosensitizing and cytotoxic effects. Suppository and intravenous 5-FU administration were compared with respect to myelo-suppression and tissue concentrations. METHODS: Rats received 120 mg/kg 5-FU-i.v. or via suppository (5 FU-S). White blood cell count, serum albumin, alkaline phosphatase, creatinine, and aspartate aminotransferase (AST) were determined before and serially after 5 FU administration. In a separate experiment, rats received 5-FU-S or 5-FU-i.v. as already described. Portal and systemic blood, rectal, iliac lymph node, liver, and lung tissue were harvested for high-performance liquid chromatography determination of 5-FU concentrations 30 min, 1, 3, 6, and 12 h after drug administration. RESULTS: No toxicity was observed in 5-FU-S animals, whereas 63% of 5-FU-IV animals had diarrhea. Weight loss and myelosuppression occurred only in 5-FU-i.v. animals. Rectal drug concentrations were significantly higher in the 5-FU-S animals compared with 5-FU-i.v. animals, 0.5-6 h after drug administration. Blood, liver, and lung 5-FU concentrations with 5-FU-S were comparable to those with 5-FU-i.v. CONCLUSIONS: 5-FU suppositories are associated with fewer systemic side effects and higher rectal 5-FU concentrations than with 5-FU-i.v. administration. PMID- 8726184 TI - Patient satisfaction with decision-making for breast cancer therapy. AB - BACKGROUND: Many sources have suggested that mastectomy is inappropriately performed too frequently for breast cancer, leading to excessive patient dissatisfaction and unnecessary mutilation. Hurried decision-making based on inadequate information has been proposed as an explanation. METHODS: After confirming the diagnosis of breast cancer, patients were informed of the diagnosis, prognosis, and treatment options according to a standard protocol. The protocol was similar to that used by many surgeons in similar circumstances. Six months after completion of either mastectomy or breast conservation therapy, patients were surveyed about their satisfaction with the decision-making process and choice of treatment. RESULTS: The majority of patients, whether they had undergone mastectomy or breast conservation, thought they had been adequately informed of treatment options and that they had made the appropriate choice of therapy. A significant percentage of mastectomy patients found that procedure more disfiguring than anticipated, but still thought they had made the appropriate choice of therapy. Despite having been informed to the contrary, most patients said their chosen treatment provided the best chance for cure. CONCLUSIONS: When informed of the diagnosis and treatment options in an unhurried, supportive setting, and when encouraged to seek further consultations as desired, breast cancer patients make appropriate therapeutic choices about mastectomy or breast conservation therapy. PMID- 8726183 TI - Primary non-Hodgkin's lymphoma of the stomach: three radical modalities of treatment in 75 patients. AB - BACKGROUND: Non-Hodgkin's lymphoma (NHL) remains a rare form of gastric malignancy, with a rising incidence. Approaches to treatment vary from surgery alone to conservative management. METHODS: To determine the optimal scheme of treatment, a randomized clinical trial was undertaken. Seventy-five patients were randomized into three groups: A-surgery alone (25), B-surgery followed by chemotherapy (29), and C-radiation therapy followed by surgery and chemotherapy (21). Forty-nine patients had stage IE and 26 had stage IIE disease. Chemotherapy (COP and COPP) consisted of 6 courses during a 1-year period, with the courses being 6 weeks apart. RESULTS: Subtotal gastrectomy was performed in 26 patients. Forty-nine patients underwent total gastrectomy. Postoperative complications occurred in 6 (8%) patients: 3 (12%) in group A, 2 (6.9%) in group B, and 1 (4.7%) in group C. Postoperative mortality occurred in 2 (8%) patients in group A (2.7% of all patients). An increase in hospital admissions number per year and decrease of mean age of patients with NHL of the stomach after the Chernobyl accident on April 26, 1986 was noted. CONCLUSIONS: Improved survival in gastric NHL was achieved by a combination of preoperative radiation with surgery and postoperative chemotherapy, presumptively through the management of local and systemic disease. PMID- 8726185 TI - Complications of mastectomy and their relationship to biopsy technique. AB - BACKGROUND: Wound complication rates after mastectomy are associated with several factors, but little information is available correlating biopsy technique with the development of postmastectomy wound complications. Fine-needle aspiration (FNA) biopsy is an accurate method to establish a diagnosis, but it is unknown whether this approach has an impact on complications after mastectomy. METHODS: Charts of 283 patients undergoing 289 mastectomies were reviewed to investigate any association between biopsy technique and postmastectomy complications. RESULTS: The diagnosis of breast cancer was made by FNA biopsy in 50%, open biopsy in 49.7%, and core needle biopsy in 0.3%. The overall wound infection rate was 5.3% (14 of 266), but only 1.6% when FNA biopsy was used compared with 6.9% with open biopsy (p = 0.06). Among 43 patients undergoing breast reconstruction concomitantly with mastectomy, the infection rate was 7.1% (0% after FNA, 12% after open biopsy). Neither the development of a postoperative seroma (9.8%) nor skin flap necrosis (5.6%) was influenced by the biopsy technique used. CONCLUSIONS: These data suggest that wound infections after mastectomy may be reduced when the diagnosis of breast cancer is established by FNA biopsy. PMID- 8726186 TI - Preoperative therapy for advanced pelvic malignancy by isolated pelvic perfusion with the balloon-occlusion technique. AB - BACKGROUND: Although the technique of isolated pelvic perfusion dates back to the time of Creech (1959) and has been used by a variety of authors to treat unresectable neoplasms, the inherent complexity of the open procedure limited its widespread use. We simplified the technique through use of the balloon-occlusion technique for aortic and caval control. Our initial efforts used this technique for unresectable pelvic cancer, but recently we used this as preoperative therapy for advanced pelvic malignancy. METHODS: Isolated pelvic perfusion was accomplished by placement of balloon-occlusion catheters (Fogerty 8) in the aorta and inferior vena cava (IVC) at L3 vertebral body level via the common femoral artery and vein and establishing inflow and outflow catheter connections to a hemodialysis pump that generated a flow rate of 150-300 ml/min. Chemotherapy drugs were infused at times 0, 10, and 20 min. 5-Fluorouracil (5-FU; 1,500 mg/M2), cis-platinum (50-100 mg/M2), and mitomycin (15-20 mg/M2) were given by normothermic perfusion over a 45-min period. Forty isolated perfusions were carried out in 25 patients. Patients were evaluated by clinical examination, biochemical tests, computed tomography (CT) and magnetic resonance imaging (MRI) scans, and surgical exploration. RESULTS: Pelvic perfusion generally achieved pelvic systemic exposure ratios (area under the curve) between 5 and 10:1 for all three drugs: mean ratios were 11.4 (5-FU), 6.0 (cisplatin), and 9.0 (mitomycin). The amount of leaking to the systemic circuit ranged from 28 to 38%. Of 15 patients treated for palliation, there was one objective partial response (PR). Ten patients had symptomatic improvement of pain, two had complete pain relief (CR), and eight had partial pain relief, ranging from 3 weeks to 3 months (median, 5 weeks). Six of 10 patients with adequate carcinoembryonic antigen (CEA) follow-up data had a reduction in CEA levels (mean change, 35 units). Of 10 preoperative patients, there was one CR among five rectal cancer patients; and four of five PRs among patients with other pelvic malignancies: two PRs in patients with epidermoid cancer and one PR each in patients with endometrial cancer and metastatic anorectal melanoma. CONCLUSION: Pelvic perfusion by a simplified balloon-occlusion technique provides palliation for most patients with advanced pelvic malignancy and may increase resectability and improve tumor control in patients amenable to resection. PMID- 8726187 TI - Complications and outcome of external hemipelvectomy in the management of pelvic tumors. AB - BACKGROUND: Although the technique of external hemipelvectomy has been adequately described, little is known about its complications and late results. DESIGN: Retrospective review of 68 external hemipelvectomies performed at our Institute between 1973 and 1994. MATERIALS AND METHODS: Eleven patients had bone tumor; 39 patients, soft-tissue sarcoma; seven patients, melanoma; 10 patients, squamous cell carcinoma; and one patient, giant neurofibroma. In 48 (71%) patients, the intent was curative. In 17 cases, the hemipelvectomy was extended. RESULTS: Postoperative complications occurred in 36 (53%) patients, including flap necrosis in 11 (16%), wound infection in 24 (35%), and other complications in 12 (18%). Four (6%) patients died postoperatively. The average hospital stay after curative versus palliative resection was 39 versus 24 days. Only three (5%) patients were able to use a prosthesis, whereas 55 (81%) used crutches, six (9%) remained wheelchair bound, and four patients (6%) spent most of the time in bed. Local recurrence occurred in 35% of the patients. The estimated 5-year survival for curatively resected patients was 21%. CONCLUSIONS: External hemipelvectomy is a procedure with considerable morbidity and is indicated for only a minority of far-advanced tumors. It offers a chance of palliation and possibly cure when lesser surgical options have been exhausted. PMID- 8726188 TI - Radioimmunoguided surgery for colorectal cancer. AB - BACKGROUND: Operations for patients with colorectal cancer are based on traditions established by historical experience. Radioimmunoguided surgery (RIGS) provides new information that challenges these traditions. METHODS: Thirty-two patients with primary colorectal cancer underwent RIGS after being injected with anti-TAG-72 murine monoclonal antibody CC49 labeled with iodine-125. Sixteen of the patients had all gross tumor and RIGS-positive tissue removed (RIGS-negative group), and 16 had only traditional extirpation of the tumor because RIGS positive tissue was too diffuse (RIGS-positive group). RESULTS: In the 16 patients having all RIGS-positive tissue removed, five had traditional regional en bloc resections and 11 had additional extraregional tissues resected. Identification of extraregional disease added two liver resections and 25 lymphadenectomies: 10 of the gastrohepatic ligament, five celia axis, six retroperitoneal, and four iliac. With a median follow-up of 37 months, survival in the RIGS-negative group is 100%. In 14 of 16 patients (87.5%) there is no evidence of disease. In the RIGS-positive group, follow-up shows 14 of 16 patients are dead and two are alive with disease (p < 0.0001). CONCLUSION: These results suggest that RIGS identifies patterns of disease dissemination different from those identified by traditional staging techniques. Removal of additional RIGS-positive tissues in nontraditional areas may improve survival. PMID- 8726189 TI - B7-1 gene transfer into human cancer cells by infection with an adenovirus-B7 (Ad B7) expression vector. AB - BACKGROUND: Transfection of the costimulatory molecule B7-1 into some murine tumors can increase antitumor immunity and eradicate tumor growth. The purpose of this work was to construct an adenovirus-B7 (Ad-B7) expression vector and study B7-1 gene transfer into human cancer cells. METHODS: The human B7-1 cDNA was ligated into an expression cassette containing the human cytomegalovirus immediate early gene promoter and then inserted into the E1 region of the Ad5 genome by homologous recombination. The resulting Ad-B7 vector was used to infect established cancer cell lines and freshly resected cancers. Resected tumors were disaggregated into single cell suspensions by mechanical mincing and enzymatic digestion. Surface expression of B7-1 after infection was verified by flow cytometry. RESULTS: Expression kinetics in three cell lines showed that infected cells began to express B7-1 within 24 h. The proportion of B7-1+ cells continued to increase during the next 48 h, after which expression remained relatively constant during the next 5 days (up to 98% B7-1+ cells). Fresh tumor cells from various cancers displayed similar kinetics, but with greater variability in the proportion of cells expressing B7-1 (13% to 95% B7-1+ cells). Cancers which were successfully infected included 3 colorectal adenocarcinomas, 2 leiomyosarcomas, 2 lung squamous cell carcinomas, and 1 renal cell carcinoma. CONCLUSIONS: The Ad-B7 vector is a rapid and efficient means of gene transfer which does not require host cell proliferation. The ultimate objective is to engineer autologous tumors to express B7-1 and vaccinate cancer patients in an adjuvant or palliative setting. PMID- 8726190 TI - The needle biopsy should replace open excisional biopsy. PMID- 8726191 TI - The Hippocratic enigma. PMID- 8726192 TI - Practice variations and the challenge to leadership. PMID- 8726193 TI - Origin of nerves supplying the posterior portion of lumbar intervertebral discs in rats. AB - STUDY DESIGN: The authors studied the origin of nerves supplying the posterior portion of lumbar intervertebral discs in rats by resection of the sympathetic trunks. OBJECTIVE: To understand discogenic low back pain from the innervation of the lumbar intervertebral discs. SUMMARY OF BACKGROUND DATA: The afferent pathways of discogenic low back pain have not been studied thoroughly. It has been reported that stimulation of an inflamed lower spinal nerve root elicits leg pain but not low back pain and that stimulation of the posterior portion of lumbar intervertebral discs evokes only low back pain. These facts suggest that pain sensation from the posterior portion of lumbar discs is not transmitted via the lower spinal nerve roots. METHODS: Forty-five Wistar rats were used. Seven days after the resection of sympathetic trunks with ganglia at different levels, the whole lumbar spine was stained by an acetylcholinesterase histochemical method. The posterior portions of lumbar intervertebral discs were observed. RESULTS: The dense nerve network on the posterior portion of lumbar intervertebral discs had disappeared almost completely after total resection of bilateral sympathetic trunks at L2-L6. However, there was a slight decrease in the network after bilateral single-level resection or unilateral multisegmental resection. CONCLUSIONS: The results showed that the posterior portion of lumbar intervertebral discs was innervated by the sympathetic nerves multisegmentally and bilaterally. PMID- 8726194 TI - An anatomic study of neuropeptide immunoreactivities in the lumbar dura mater after lumbar sympathectomy. AB - STUDY DESIGN: The distribution and density of nerve innervation in the lumbar dura mater after lumbar sympathectomy were assessed in wistar rats. OBJECTIVE: To provide basic information on the interaction between sympathetic and sensory nerves in patients with low back pain. SUMMARY OF BACKGROUND DATA: Many studies have indicated that the sinuvertebral nerve has an important role in innervating the tissues around the vertebrae. However, the origin, innervating pattern, and connections between the nerves are still controversial. It is well known that pain is often accompanied with sympathetic symptoms and exaggerated by sympathetic stimuli. Occasionally, anesthetic block at the L2 or L3 sympathetic ganglion relieves low back pain or symptoms associated with low back pain. The authors assessed the changes of the density and distribution of nerve innervation of the lumbar dura mater after lumbar sympathectomy. METHODS: Normal adult rats were sympathectomized at L2-L3. The threshold for thermal noxious pain by hot plate analgesia test and changes in neuropeptides in the lumbar dura mater and dorsal root ganglia using light microscopic immunohistochemistry were assessed and compared with control rats. RESULTS: In the hot-plate analgesia test, sympathectomized rats increased their hot-plate latency time compared with that of sham-operated rats. Density of calcitonin gene-related peptide immunoreactive fibers in sympathectomy side of the lumbar dura mater decreased to 45.5% compared with the contralateral side. The number and size of calcitonin gene-related peptide immunoreactive cells in dorsal root ganglia showed no difference between sympathectomized and contralateral side. CONCLUSION: Sympathectomy increased the pain threshold and made the sympathectomized rats hypesthetic. A large numbers of sensory fibers innervated the lumbar dura mater via L2-L3 sympathetic nerve in rats. Sympathectomy reduced the number of these nerve fibers in the lumbar dura mater. Sympathetic nerves may play an important role for low back pain involving the lumbar dura mater. PMID- 8726195 TI - Neurophysiologic changes in lumbar nerve root inflammation in the rat after treatment with cytokine inhibitors. Evidence for a role of interleukin-1. AB - STUDY DESIGN: The present study was designed to evaluate the effect of cytokine inhibitors in experimental allergic radiculitis. OBJECTIVE: Evaluation of the effect of cytokine inhibitors in experimental allergic radiculitis. SUMMARY OF BACKGROUND DATA: A number of cytokines are known to be involved in hyperalgesia and may play a role in radiculitis. Corticosteroids and other cytokine inhibitors antagonize their effects. METHODS: Experimental allergic radiculitis was induced in rats by injection of bovine myelin from the peripheral nervous system. The sham group subsequently received saline injections; the treatment groups received either prednisolone or interleukin-1 receptor antagonist. Treatment effect was assessed on the basis of motor performance and neurophysiologic parameters. RESULTS: Treatment ameliorated the symptoms of experimental allergic radiculitis. Prednisolone appeared to be somewhat more effective than interleukin-1 receptor antagonist. CONCLUSIONS: Because interleukin-1 receptor antagonist specifically blocks the effects of interleukin-1 at its receptors, the present results imply that interleukin-1 is a causal factor in the model of experimental radiculitis used. Its specificity and apparent lack of side effects make interleukin-1 receptor antagonist an attractive candidate treatment for the human disease. PMID- 8726196 TI - Remodeling of vertebral bone after outer anular injury in sheep. AB - STUDY DESIGN: This study uses semiautomated histoquantitative techniques to analyze subchondral bone architecture in adult merino sheep up to 2 years after outer anular cuts were created in lumbar discs. OBJECTIVES: To describe morphometric changes to the adjacent vertebral bone structure in the lumbar spine, and to discuss these in relation to the pathology of other spinal components SUMMARY OF BACKGROUND DATA: Disc degeneration has been observed in the sheep lumbar spine 4-6 months after surgery to the outer anulus, whereas biomechanical testing has shown earlier changes. METHODS: Lumber discs with adjacent subchondral trabecular bone were sectioned using standard histologic methods. An image-analyzing computer was used to measure morphometric indices of bone structure. RESULTS: Trabecular bone volume was elevated significantly by 2 months because of consolidation of existing trabeculae and formation of new elements. The changes were evident 2 years after surgery. CONCLUSION: Subchondral trabecular bone appears to be sensitive to changes in the disc after outer anular injury. There is evidence that these structural changes are influenced by changes in the biomechanical behavior of the incised disc. PMID- 8726197 TI - Back muscle injury after posterior lumbar spine surgery. A histologic and enzymatic analysis. AB - STUDY DESIGN: Back muscle injury after posterior lumbar surgery was studied by muscle histology and serum creatine phosphokinase MM isoenzyme activity. OBJECTIVES: To investigate intraoperative factors influencing the magnitude of back muscle injury after posterior lumbar surgery. SUMMARY OF BACKGROUND DATA: The authors previously have reported iatrogenic back muscle injury in an animal model and in humans. Serious injury of the back muscle has been shown by short term and long-term follow-up evaluation. METHODS: The retraction pressure was monitored, and the retraction pressure-time products were calculated in 24 patients. Early histologic changes of multifidus muscle, which were taken at completion of surgery, and serum creatine phosphokinase MM isoenzyme activity changes were examined. RESULTS: The magnitude of back muscle injury was significant as the pressure-time product increased. Creatine phosphokinase MM isoenzyme activity increased after surgery and reached a plateau 1 day after surgery, followed by recovery to the normal value 1 week after surgery. Creatine phosphokinase MM isoenzyme activity tended to be high in cases with multilevel exposure and with high pressure-time product. CONCLUSIONS: Back muscle injury occurs in all patients who underwent posterior lumbar surgery, and these injuries are related to the retraction pressure, time, and extent of exposure. PMID- 8726198 TI - Early prediction of chronic disability after occupational low back injury. AB - STUDY DESIGN: An inception cohort design was used to study a consecutive sample of back-injured workers. OBJECTIVE: To refine and to test the Vermont Disability Prediction Questionnaire's ability to indicate an individual's relative risk for chronic disability after occupational low back injury. SUMMARY OF BACKGROUND DATA: Although most back-injured workers return to work quickly, the minority who do not account for the majority of associated costs and health care. Early identification of workers at high risk for disability would facilitate intervention strategies. METHODS: During the study recruitment period, people aged 18-60 years reporting occupational low back injury to the Vermont Department of Labor and Industry within 11 days of onset were eligible. A Vermont Disability Prediction Questionnaire was mailed to the 442 subjects who could be contacted and who gave informed consent. One hundred sixty-three of the 166 subjects who completed and returned the questionnaire within 15 days of initial injury were telephoned 3 months later to determine work status. Those who returned the questionnaire were compared with those who would not consent or did not return the questionnaire in time according to age, sex, residence, wages, work hours per week, and length of employment. Employment status (inability to work because of low back pain) was evaluated by telephone interview 3 months after initial injury. RESULTS: The follow-up interviewer was blinded to the Vermont Disability Prediction Questionnaire scores. of the 163 subjects, 16 (10%) were not working because of low back pain. Using a simple dichotomous scoring system for 11 questionnaire items, a cut-off score of 0.48 identified 3-month postinjury work status, with 0.94 sensitivity and 0.84 specificity. CONCLUSIONS: The Vermont Disability Prediction Questionnaire is a brief, easily administered and scored tool for identifying back-injured workers at relative risk for chronic disability. Such early identification should increase the efficiency of disability prevention strategies by directing them toward people who need them most. The accuracy of the questionnaire needs to be tested in a variety of different clinical and socioeconomic settings. PMID- 8726199 TI - Recurrent or new injury outcomes after return to work in chronic disabling spinal disorders. Tertiary prevention efficacy of functional restoration treatment. AB - STUDY DESIGN: A large prospective longitudinal cohort study (n = 1204) to identify prevalence of new or recurrent injury and risk factors in a rehabilitated chronic disabling spinal disorder patient group with matched control subjects. OBJECTIVES: To evaluate prevalence and risk factors for new or recurrent injury on a chronic disabling spinal disorder population. SUMMARY OF BACKGROUND DATA: The rate of symptom recurrence after acute low back pain, like the rate of initial back pain episodes, is extremely high (40-70%). However, although the incidence of recurrent back pain after chronic disabling spinal disorder represents a small subcomponent of these cases, there is a large socioeconomic impact. An individual attempting to return to work after a chronic disabling spinal disorder episode usually bears a stigma of "high risk" for recurrent injury and related work disability that may result in barriers to reemployment or work retention. Before the present research, no large scale studies had been conducted to evaluate whether recurrent spine injuries or new injuries to other musculoskeletal areas could be prevented by medical treatment. In addition, no studies had emerged to evaluate physical and psychologic risk factors of injury recurrence in this context to facilitate design of prevention programs. METHODS: The present study assessed the incidence of claimed recurrent spinal and new musculoskeletal injuries in a population of 1204 workers. A subgroup of 5.3% (n = 64) of treated patients with a new injury claim in the ensuing 12 months was matched for gender, age, race, length of disability, workers' compensation venue, previous surgery, and litigation status to an identically sized control group who did not report new or recurrent injuries. Demographic, physical, and psychologic measures were obtained prospectively on all patients, before and after treatment, to be analyzed as risk factors. RESULTS: During the year after treatment, 1.3% (n = 16) of patients reported another injury to the same spinal area, with only a 0.9% (n = 11) recurrent disability rate. A new injury to a different musculoskeletal area was reported by 4.0% (n = 48) of patients. Only 3.4% of the whole cohort, or 64.1% (n = 41) of the sample reporting reinjuries after returning to work, experienced lost work time (i.e., disability) after the reinjury. Only a modest predictive association was found between risk for new or recurrent injury and two self-report indices. CONCLUSIONS: The present study suggests that even a sample of the most severe chronic disabling spinal disorder workers' compensation patients who complete a tertiary functional restoration program are at relatively low risk for either a recurrent spinal disorder or new musculoskeletal injury claim (with or without disability). No major physical or psychologic risk factors for recurrent injury could be identified in this large cohort. These findings argue powerfully against employer bias in not rehiring employees with previous chronic disabling spinal disorder or discriminating in pre- or reemployment on the basis of putative reinjury risk factors after an appropriate rehabilitation program. Literature review documents a surprising paucity of quality studies examining variables predictive of this important socioeconomic outcome variable. PMID- 8726200 TI - Health outcome assessment before and after lumbar laminectomy for radiculopathy. AB - STUDY DESIGN: Forty-one patients undergoing lumbar laminectomy for radiculopathy resulting from herniated discs assessed their health status using a generic health outcome instrument (Medical Outcomes Study Short Form 36) before surgery and at an average of 2 years after surgery. OBJECTIVES: To assess whether lumbar laminectomy for herniated nucleus pulposus is a useful intervention when patients evaluate their own perception of health. SUMMARY OF BACKGROUND DATA: The medical Outcomes Study Short Form 36 has been used in multiple studies assessing various medical conditions. It is brief, generic, and reliable. Although surgical treatment for radiculopathy by lumbar laminectomy has been shown to be successful using specific criteria for patient selection and an algorithmic approach, the authors are not aware of any study using a patient-based health outcome assessment to evaluate the results of this type of surgery. METHODS: Forty-one patients (82% completed follow-up evaluation; average follow-up period, 2.08 years) completed Medical Outcomes Study Short Form 36 before and after surgery. Scores from before and after surgery were compared. RESULTS: Statistically significant improvements (P < 0.01) were seen in eight of the nine health scores comparing scores from before and after surgery at follow-up evaluation. These included physical function, social function, role function resulting from physical limitations, role function resulting from emotional limitations, mental health, vitality, pain, and perceived health change. No significant change was seen in the patients' health perception after surgery. CONCLUSIONS: This study shows that the patients' self-reported health outcomes after lumbar laminectomy correlate with the excellent results previously seen using physician-driven outcome measures in an appropriately selected population with radiculopathy. The excellent results shown here did not deteriorate with age (> 40 years compared with < 40 years) or with complications after surgery. PMID- 8726201 TI - Lumbar lordosis in spinal fusion. A comparison of intraoperative results of patient positioning on two different operative table frame types. AB - STUDY DESIGN: One hundred one patients undergoing spine surgery for degenerative conditions were entered into a prospective radiographic evaluation of changes in lumbar lordosis as affected by positioning on two different operative tables. OBJECTIVES: The hypothesis of the present study is twofold: 1) the positioning of patients on specific types of operative tables may affect significantly the overall degree of lumbar lordosis obtainable, and 2) certain operative positioning may more accurately reproduce physiologic standing lateral lumbar lordosis. SUMMARY OF BACKGROUND DATA: In the management of degenerative and post traumatic spinal deformities, lumbar fusion using posterior instrumentation permits more accurate and physiologic lordotic positioning of the involved fusion segments of the lumbar spine. However, various types of operating frames are available for use in this type of surgery, and despite the overall importance of correct lordotic positioning, there is some question as to what effect on positioning, as measured in degrees of lumbar lordosis, a particular frame might have. METHODS: Total, multisegmental, and unisegmental Cobb angle measurements of preoperative standing lateral radiographs and intraoperative lateral radiographs after positioning on respective operative tables were determined. Fifty-one patients were positioned on an Andrews-type table, and 50 patients were positioned on the four-poster-type frame. Statistical comparison using analysis of variance testing of changes in lordosis before and after surgery between study groups was evaluated. RESULTS: Lumbar lordosis measured from L1 to S1 with standing lateral radiographs showed a combined mean preoperative measurement of 45.18 degrees, with no statistical significance between groups. In comparison, there was a statistically significant difference between intraoperative measurements from L1 to S1 on the Andrews table versus the four-poster frame, revealing an average of 32.81 degrees versus 47.71 degrees, respectively (P < 0.005). Multisegmental lordosis measurement from L2 to S1 displayed statistical significance between groups, with a combined preoperative standing lateral radiograph average of 43.32 degrees, and intraoperative values of 31.28 degrees on the Andrews table versus 45.34 degrees on the four-poster frame (P < 0.005). Multisegmental lordosis measurements from L4 to S1 displayed statistical significance between groups, with a combined preoperative standing lateral radiograph average of 31.40 degrees and intraoperative values of 23.14 degrees on the Andrews table versus 32.94 degrees on the four-poster frame (P < 0.005). Segmental lordosis at L5-S1 was less dependent on frame type, with a combined preoperative standing lateral radiograph average of 20.53 degrees and intraoperative measurements of 20.06 degrees on the Andrews table versus 21.02 degrees on the four-poster frame (P < 0.43). CONCLUSION: Results from the present study display a statistically significant difference between multisegmental and total lumbar lordosis, depending on the type of operative table used in patient positioning. Segmental lordosis at L5-S1 depended less on frame type. This table dependent positional change in lumbar lordosis could be incorporated easily into a lumbar fusion procedure, especially when supplemented with instrumentation, affecting the permanent overall degree of lordosis. These results suggest that a more physiologic degree of lumbar lordosis is obtained accurately with use of an operative table similar to the four-poster frame. PMID- 8726202 TI - Lumbar motion segment pathology adjacent to thoracolumbar, lumbar, and lumbosacral fusions. AB - STUDY DESIGN: Fifty-eight patients came to two spinal surgeons with abnormalities adjacent to a previously fused thoracic or lumbosacral segment after they had been asymptomatic for more than 2 years. Radiographs, outcome analysis, analog pain evaluation, patient demographics, and medical indices were analyzed to evaluate risk factors for adjacent segment abnormality. OBJECTIVES: Risk factors for adjacent segment breakdown and pathophysiology of adjacent segment breakdown were analyzed in this longitudinal study. SUMMARY OF BACKGROUND DATA: Fifty-eight patients underwent a thoracolumbar, lumbar, or lumbosacral fusion with an average symptom-free period of 13.1 years before presentation with severe symptomatology necessitating further surgery at the adjacent segment. METHODS: Fifty-eight patients with adjacent segment abnormality were analyzed by outcome assessment questionnaire, pain analog evaluation, radiographic studies, demographic factors, and sequential follow-up evaluation. Thirty-seven of these patients have been followed for more than 2 years after their adjacent segment surgery. RESULTS: Fifty-eight patients developed spinal stenosis, disc herniation, or instability at a segment adjacent to a previously asymptomatic fusion that was done an average of 13.1 years earlier. Segments adjacent to the adjacent segment itself were as likely to breakdown (58%). Thirty-seven patients were followed for more than 2 years, having outcomes defined as good or excellent in 70.3%. Seven of the 37 patients required an additional surgical procedure. Sagittal and coronal imbalances appeared to play a role in breakdown, although statistical significance was not evident. CONCLUSIONS: This represents the largest series of adjacent segment breakdowns reported in the literature. The segment adjacent to the adjacent segment was almost as likely to breakdown. Sagittal and coronal alignment appeared to play a role in adjacent abnormality. Good outcomes are evident in 70% of cases. PMID- 8726203 TI - The safety and efficacy of the Isola Spinal Implant System for the surgical treatment of degenerative disc disease. A prospective study. AB - STUDY DESIGN: This is a prospective study designed in consultation with and approved by the Food and Drug Administration with the purpose of determining the safety and efficacy of the Isola Spinal Implant System for the surgical treatment of patients with degenerative disc disease. OBJECTIVES: To report the results of the degenerative disc disease group from the Isola Investigational Device Exemption study, which was done to determine whether the Isola Spinal Implant System is a safe and effective treatment. SUMMARY OF BACKGROUND DATA: The safety and efficacy of transpedicular instrumentation as an adjunct in achieving lumbar spine fusion are still debated. METHODS: Hospital Investigational Review Board approval of the study protocol was obtained at the 10 participating centers. One hundred twenty patients (49 men and 71 women; average age, 54 years [range, 25-83 years]) were enrolled. Clinical and radiographic follow-up evaluation was done using protocols established prospectively. RESULTS: Of the 120 patients, 12 (10%) had device-related problems. There were two operative and four device related complications after surgery. Six other patients had their implants removed either for local pain (two) or for looseness or breakage (four). Of 118 patients eligible for follow-up evaluation at 24 months, 107 (91%) were available for study. Fusion was achieved in 97 (91%) patients. Average combined function and pain scores improved by 2.6 points (P < 0.0001). Clinical success, as measured by combining function and pain scores, was achieved in 65% of the patients by the most stringent criteria and 73% by less stringent criteria. Clinical success was significantly higher in patients who had not had previous surgery than in those who had, 77% versus 57% (P = 0.04). CONCLUSIONS: The present study suggests that the Isola Spinal Implant System can be safely used, is an effective adjunct in the achievement of fusion, and yields an acceptable number of successful clinical outcomes, especially when considering the preponderance of previously operated patients included in this group. PMID- 8726204 TI - Results of disc prosthesis after a minimum follow-up period of 2 years. AB - STUDY DESIGN: This study analyzed retrospectively 46 patients undergoing artificial disc replacement. OBJECTIVE: To assess the clinical and radiographic outcomes of patients who had Charite SB III disc prosthesis. SUMMARY OF BACKGROUND DATA: Results of disc prosthesis have been reported only at very short term follow-up periods; preoperative diagnosis and criteria used to evaluate the clinical outcomes were not reported. METHODS: Forty-six patients who had had a disc prosthesis were evaluated clinically and radiographically at least 2 years after surgery. Preoperative diagnosis included disc degeneration in 22 patients and failed disc excision in 24 patients. Disc prosthesis was implanted at a single vertebral level in 36 patients and at two levels in 10 patients. Follow-up evaluation was performed after an average of 3.2 years (range, 2-5 years). RESULTS: Sixty-three percent of patients reported satisfactory results. The success rate was 69% in patients who underwent isolated disc replacement and 77% in those with no previous back surgeries. Seven patients who had unsatisfactory results underwent posterolateral fusion without removing the artificial disc. Two patients underwent removal of the prosthesis. No failure of the implants or loosenings or wear of the polyethylene core were found. Vertebral motion averaged 9 degrees at the operated levels and 16 degrees at the adjacent levels. CONCLUSION: A wrong surgical indication, rather than failure of the prosthesis, appears to be the main cause of unsatisfactory results of disc replacement at medium-term evaluations. Prospective and longer term studies are needed to establish whether disc prosthesis may offer advantages compared with spinal fusion. PMID- 8726205 TI - Intraoperative chemonucleolysis as an adjunct to lumbar discectomy. PMID- 8726206 TI - Keeping care in health care reform. PMID- 8726208 TI - Practice patterns of rural family physicians based on the American Diabetes Association standards of care. AB - The purpose of this study was to examine practice patterns of rural family physicians in the care of non-insulin-dependent diabetes mellitus based on the standards of care of the American Diabetes Association (ADA). One hundred patient charts were randomly chosen, twenty for each physician, from the practices of five family physicians in rural Ohio. A standardized collection protocol was used, based upon the ADA recommendations. The charts were reviewed for compliance with the ADA parameters. The patients' records demonstrated 66% compliance with dietary counseling and 33% with counseling about exercise. Moreover, there was low compliance with physical examination guidelines. Specifically, 66% of the patients had fundoscopic examination and 64% had a complete foot examination done. With respect to the laboratory guidelines, 70% of the charts reviewed had a urinalysis ordered and 45% annual lipids measured. However, glycosylated hemoglobin was performed in only 15% of the patients. The results suggest that rural family physicians do not consistently follow the ADA standards of care. PMID- 8726207 TI - Continuity of care and delivery of ambulatory services to children in community health clinics. AB - This study assesses how continuity of care influences receipt of preventive care and overall levels of ambulatory care among children and adolescents in community health clinics (CHCs). It is a secondary data analysis of the 1988 Child Health Supplement to the National Health Interview Survey. Of 17,110 children in the sample population, the 1465 who identified CHCs as their routine source of care formed the study population. Continuity of site was defined as identification of a CHC as a source of both routine and sick care, and continuity with a clinician was defined as identification of a specific clinician for sick visits. In bivariate analyses both continuity with the CHC and with a specific clinician were associated with increased levels of preventive care and overall ambulatory care. In logistic regression models, continuity of care was associated with nearly a two-fold increase in the odds of receiving age-appropriate preventive care. Alternatively, insurance status was a better predictor of receipt of overall levels of ambulatory care. We conclude that expanding financial access alone is unlikely to sufficiently improve low-income children's access to Community Health Clinics. Additional emphasis on localizing the delivery of both routine and sick care services in a single site or with a specific clinician may be needed to achieve higher levels of both preventive care and overall ambulatory care. PMID- 8726209 TI - Self-perceived unmet health care needs of persons enrolled in HIV care. AB - We examined the prevalence of, and factors associated with unmet health service needs among persons with HIV disease. Data were examined from 1,851 participants in the U.S. AIDS Cost and Service Utilization Study, drawn from 26 medical care providers in 10 cities. Geographic areas with large numbers of AIDS cases, and health care providers within them were chosen as study sites. After completing a screener questionnaire, potential participants at each site were stratifed by illness stage, HIV exposure route, and insurance status; a systematic random sample within those strata were selected for the study. Participants completed a comprehensive survey of HIV-related service use and costs, which also asked them to identify unmet health service needs. Analyses identified the relationship between unmet needs and: stage of illness, type of insurance, source of care, living arrangement, and AIDS prevalence of respondents' geographic region. At least one unmet need was reported by 20% of the sample. Needs for non institutional services, e.g, dental care, mental health, and medications were more likely to be unmet than need for emergency room and hospital care. While most factors significantly affected the odds of having an unmet need, the greatest effects were found for private insurance and HIV asymptomatic status, both of which decreased the odds of unmet needs by approximately 50%. These findings suggest that insurance coverage for services required during the chronic phase of HIV illness is inadequate and should be augmented. PMID- 8726210 TI - The impact of the Medicare Influenza Demonstration Project on influenza vaccination in a county in Massachusetts, 1988-1992. AB - Influenza and related pneumonia continue to cause significant amounts of morbidity and mortality despite the availability of effective vaccines. Two comparable counties in Massachusetts served as project areas of a national trial to see if reimbursement for immunizing Medicare-B eligible recipients against influenza would increase the use of the vaccine and reduce the costs attributed to related morbidity. Providers of health and social services to elders were recruited to participate in one county. A variety of professional and public education campaigns and media were used to promote influenza immunizations. Laboratory-based surveillance was instituted in both counties to assess the extent of circulating virus in each. Vaccine was made available to medical providers in both counties. While the amount of vaccine used in the comparison county increased by 6% from pre-project time (16,000 to 17,000 doses administered), vaccine use increased 219% in the intervention county (21,250 to 46,494 doses administered). In a post-project survey of participating physicians, 88% of 238 respondents reported administering less than 100 doses of influenza vaccine per year prior to the project. By the end of the project, only 32% administered less than 100 in the previous year. This project demonstrated the need for educating both the provider and the public in order to successfully promote immunizations. It was not clear, however, if reimbursement was a more important factor for promoting influenza immunizations than was universal distribution of free vaccine. PMID- 8726211 TI - Mary Edwards Walker, M.D.: a feminist physician a century ahead of her time. AB - In her teens, Mary Edwards Walker already wore the "bloomer" outfit began to campaign for reforming the "unhygienic" clothing of women. Assertively, she attended medical school and earned her M.D. degree. Due to prejudice, her practice did not flourish and she moved to Washington to offer her medical services to the Union as the Civil War began. Rebuffed by the male medical bureaucrats, she volunteered her services anyway. Eventually, she was awarded the Congressional Medal of Honor, the only women to ever gain such distinction. After the war, Walker became a journalist, an author of two sensational books, a political lobbyist, a suffrage campaigner, a professional and public lecturer, an ardent dress reformer, a peace activist, a Utopianist and a women's right advocate. Light-years ahead of her times, Dr. Walker was an intelligent, independent, irrepressible and indefatigable proponent for a host of worthy causes. PMID- 8726212 TI - Autoantibodies to specific enzymes: a review. AB - There are two categories of autoantibodies to specific enzymes: immunoglobulin complexed enzymes and circulating autoantibodies directed to enzymes in tissue or tissues. Immunoglobulin-complexed enzymes may result in elevated serum enzyme activity. They are found more frequently in elderly patients and have limited clinical significance. Immunoglobulin association with the enzyme must be demonstrated to distinguish this macroenzyme from other high molecular weight enzyme complexes. Autoantibodies to specific enzymes or regulators of enzyme activity do possess specific disease associations. The titers or presence of these autoantibodies may predict morbidity or response to therapy. These autoantibodies may be detected by Western blotting, enzyme-linked immunosorbent assays, tissue immunofluorescence, radioimmunoassay, immunoprecipitation flow cytometry or inhibition of enzyme activity. For example, anti-pyruvate dehydrogenase inhibits the activity of purified enzyme, but not relatively intact mitochondrial preparations. Most evidence suggests that the production of autoantibodies to specific enzymes represents an epiphenomenon secondary to tissue damage rather than a primary event in the pathogenetic pathway. PMID- 8726213 TI - Breast cancer cytogenetics: a review and proposal for clinical application. AB - Cytogenetic analyses of hematological neoplasms have proved to be relevant in the diagnosis, treatment, and prognosis of afflicted patients. Breast cancer (BC) cytogenetics is expected to contribute in the same fashion. Literature has shown that clone-specific chromosomal changes do occur in BC, yet, their clinical significance is unknown. Most of the studies have been conducted on patients with advanced stage tumor. Karyotypic analyses of the few reported cases from stage I BC tumors revealed a higher frequency of single clonal abnormalities. This work describes an ongoing BC cytogenic study on samples from stage I tumors to enhance and clarify this observation. Included as control are chromosomal analyses of peripheral blood and peritumoral normal tissue samples of these patients, which might provide information regarding predisposing cytogenetic aberrations. Non random chromosomal abnormalities in BC include involvement of chromosomes 1, 3, 6, 11, 16, and 17. Three groups of non-random chromosomal alterations, ranging from specific abnormalities, partial monosomies, and secondary changes (eg, numerical loss of chromosomes) are described. Survival appears to be more favorable in patients without complex karyotypes. A better understanding of the clinical and etiologic implications of BC is expected to emerge from continued assessment of breast tumor cytogenetics. PMID- 8726214 TI - Early breast cancer, diet, and plasma copper fractions. AB - In a study of diet and early breast cancer, blood plasma copper has been analyzed by Proton Induced X-ray Emission analysis as both total copper (P-Cu) and that ultrafiltrable from plasma (P-edu-Cu) through membranes with a cut-off at molecular mass 10,000 after equilibration with disodium ethylene diamine tetraacetic acid (EDTA) at 4 degrees C. Ceruloplasmin (P-cer) was also measured using nephelometry of anticeruloplasmin monoclonal antibody-ceruloplasmin complexes. Dietary copper intake per day (D-Cu) was assessed over a five-day dietary record period and calculated from dietary components using a computer program. P-edu-Cu correlated significantly with both D-Cu and ceruloplasmin while P-Cu correlated only with ceruloplasmin. Further, ceruloplasmin did not significantly correlate to D-Cu. Hence, P-edu-Cu better reflects copper status than do P-cer or P-Cu as it relates to both the major copper enzyme in plasma and to daily copper intake. This may be important in drawing conclusions about the significance of copper in disease states where copper fractions other than ceruloplasmin may be most important owing, for example, to oxidative properties. Categorization as cancer or normal, by copper parameters (D-Cu, P-edu-Cu, P-Cu, P cer), was studied in multiple correlation. In particular, the ratio P-cer/P-Cu and the ratio P-edu-Cu/D-Cu were significantly related to disease. Irrespective of age (pre- and post-menopausal), highly significant differences between normals and early stage breast cancer patients were seen with p < 0.0001 to p < 0.01. The precise role played by plasma and dietary copper fractions deserves continued attention in view of the present and earlier results in cancer studies. PMID- 8726215 TI - The role of silicic acid in the renal excretion of aluminium. AB - The chemical affinity of silicic acid for aluminium (AI) has been shown to reduce the bioavailability of AI in studies of human gastrointestinal (GI) absorption. Investigations were carried out to ascertain whether or not similar interactions may also enhance the renal excretion of AI by assessing the urinary output of both elements. Healthy individuals given monosilicic acid as naturally found in beer, excreted the majority of the silicic acid content (mean 56 percent) within 8 hours, concomitant with a significant increase in AI excretion (P < 0.05). Ingestion of increasing doses of silicic acid resulted in dose related increases in excretion of Si. Excretion of AI reached a maximum and then declined, consistent with depletion of AI body stores. This was confirmed using the 26AI isotope. The low serum but high urine concentration of Si suggests that if AI and Si interact to form an excretable species they do so in the kidney lumen such that Si limits the reabsorption of AI. Silicic acid's effect on the depletion of aluminium stores and reduced GI absorption suggest its addition to municipal water supplies may be a low risk public health measure to reduce the AI burden in the general population. PMID- 8726216 TI - Sphingomyelinase inhibits in vitro Leydig cell function. AB - Activation of the immune system has profound effects on endocrine function which are mediated by cytokines including tumor necrosis factor-alpha (TNF alpha). In vitro, TNF alpha has been shown to directly inhibit Leydig cell testosterone (T) production, but the mechanism of this effect is still unclear. Recent studies using cultured human fibroblasts have shown that TNF alpha stimulates the activity of neutral sphingomyelinase (SMase) which hydrolyses sphingomyelin (SM) generating ceramide and changing membrane components including cholesterol. The cellular affects of increased SMase activity have been reproduced in vitro by the addition of exogenous SMase. In cultured fibroblasts, exogenous SMase decreases cholesterol synthesis. These findings led us to hypothesize that SMase might be important in the regulation of steroid hormone synthesis. To our knowledge, no previous studies have investigated this possibility. To test this hypothesis, rat Leydig cell enriched cultures were incubated in media containing SMase (0.1 to 100 mU/ml) or in control media. SMase significantly decreased basal and human chorionic gonadotropin (hCG) stimulated T production. SMase also decreased hCG binding and hCG stimulated adenosine 3':5'-cyclic monophosphate (cAMP). N-acetyl sphingosine (0.1 to 10 microM), a water soluble ceramide, was used to determine whether or not the effects of SMase could be reproduced by ceramide addition. N acetyl-sphingosine had only slight effects on basal T and cAMP, and no effect on hCG binding or hCG stimulated T or cAMP. These data suggest the metabolism of membrane sphingomyelin may be an important regulatory pathway in the control of Leydig cell function. PMID- 8726217 TI - Immunoperoxidase staining of tumors by an antibody to Xenopus pNiXa. AB - pNiXa, a serpin from oocytes and embryos of Xenopus laevis, was tested as a tumor marker in human and rodent tissues. A peptide corresponding to the histidine-rich domain of pNiXa was conjugated and administered to rabbits to produce a polyclonal antibody, which was purified by antigen-affinity and used for immunoperoxidase staining of formalin-fixed, paraffin-embedded tissue sections. Staining with pNiXa-antibody was positive in 23/187 human tumors (12 percent) and negative in 119 specimens of normal human tissues. Positive reactions were more frequent in liver (38 percent) and colon (34 percent) tumors than breast (18 percent), prostate (9 percent), mesothelioma (20 percent) or lung (0 percent) tumors. Staining was negative in human tumors from other sites. Rodent tumors and preneoplastic foci induced by chemical carcinogens were surveyed for staining with pNiXa-antibody. Staining was positive in 10/10 hepatic lesions (hepatocellular foci, adenomas, carcinomas) induced in hybrid D2B6F1 mice by diethylnitrosamine and phenobarbital, whereas murine mammary tumors and thyroid, pituitary, renal, and colon tumors of F-344/CNr rats were negative. Thus, immunostaining with pNiXa-antibody identifies a subset of human and murine tumors; further studies are needed to determine if reactivity of pNiXa-antibody has diagnostic or prognostic significance. PMID- 8726218 TI - Involvement of analytical chemistry in chemical speciation of metals in clinical samples. AB - The different chemical species of the trace elements in a living system are determinants for their physiological behaviour. Their study is necessary to improve the understanding of trace element kinetics and metabolism. In a complex matrix, such as biological fluids and tissues, some trace elements will occur as free or mononuclear ions; other as low molecular weight complexes, as reversible or irreversible macromolecular complexes. Speciation investigations entail the separation of the compounds, followed by the measurement of the trace element in the different fractions. Frame-work-procedures are outlined and attention is drawn on the many difficulties that can be encountered. These include the complexity of the matrix, insufficient specificity of the separation of biocompounds, fortuitous contaminations with trace elements, and cutting the original metal-protein binding. State of the art description is given for the speciation studies of AI, As, Cd, Cr, Co, Cu, Hg, Ni, Pb, Pt, Se, Sn, and Zn. PMID- 8726219 TI - Metal-induced dimerization of Cd7-metallothionein. Role of anions. AB - Inorganic phosphate participates in the nonoxidative Cd-dependent dimerization of Cd7-metallothionein (Cd7-MT) and is bound to the MT dimers. In order to obtain insight into the specificity of phosphate-MT interaction, an investigation has been made on the effect of a series of oxyanions and organic phosphates on Cd induced dimerization of Cd7-MT. It has been demonstrated that from the oxyanions studied, only arsenate and tungstate can promote Cd-induced dimerization of Cd7 MT. Effect of arsenate is quantitatively similar to that of phosphate and appears at submillimolar concentrations, whereas tungstate promotes dimerization of MT at higher, millimolar concentrations. A number of other oxyanions, ie, molybdate, vanadate, and selenate as well as organic phosphates (phenyl phosphate, serine O phosphate, D-ribose 5-phosphate, L-glycerol 3-phosphate, D-fructose 6-phosphate, D-glucose 6-phosphate, adenosine monophosphate (AMP), cyclic adenosine monophosphate (cAMP), nicotinamideadenine-dinucleotide (NADP), pyridoxal 5 phosphate) had no influence on Cd-induced dimerization of MT. Results obtained strongly indicate a highly specific role of inorganic phosphate in Cd-induced dimerization of Cd7-MT. PMID- 8726220 TI - Douglas-fir: a third wood-drying condensate found to exhibit in vitro cytotoxicity and genotoxicity. AB - The present study applies the approach described in Mark et al. for the testing of toxic chemicals produced during the drying of Douglas-fir. The genotoxic potential of Douglas-fir condensate has been previously unexplored and is thus an area of appropriate concern to the forest products industry, regulatory, agencies, and the general public. Previous research conducted in this laboratory has identified two wood-drying condensates that yield positive cytotoxic and genotoxic effects. The results of testing Southern yellow pine and Eastern white pine condensates have been reported elsewhere. Douglas-fir condensate, a third wood-drying condensate, was added in vitro in concentrations ranging from 0.1 to 100 microliters/ml to cultures of Chinese Hamster Ovary (CHO-WBL) cells. A dose response curve was observed with this condensate for both cytotoxicity and genotoxicity. The number of viable cells as well as the mitotic index (MI) and proliferative rate index (PRI) varied inversely with dosage. The result of chromosome aberration (Abs) analysis and sister chromatid exchange (SCE) analysis, both cytogenetic measures of genotoxicity, also gave statistically significant results. PMID- 8726221 TI - Amino acid concentration in dementia of the Alzheimer type and multi-infarct dementia. AB - Amino acids were measured in nine cases of dementia of the Alzheimer type, 10 cases of multi-infarct dementia, and 10 healthy controls. The severity of dementia was examined using mini-mental state test (MMST). Amino acid analysis (41 kinds) in the cerebrospinal fluid (CSF) and serum was performed in the Special Reference Laboratories. In the dementia of the Alzheimer type group, methionine and alanine concentrations in the CSF were significantly increased, and the CSF/serum ratios for both the alanine and glycine concentrations were significantly increased, in comparison with the healthy control group. In the multi-infarct dementia group, glycine, methionine, threonine, phenylalanine, and citrulline concentrations in the CSF were all higher than in the healthy control group. Significant negative correlations were found between the MMST score and the alanine, urea, arginine, and alpha-aminobutyric acid concentrations in the CSF. The number of amino acids which exhibited abnormality in dementia of the Alzheimer type and multi-infarct dementia was greater in the present study than in previous reports. PMID- 8726222 TI - Acquired free protein S deficiency in children with steroid resistant nephrosis. AB - Plasma and urine concentrations of protein S were measured in five children with steroid-resistant nephrotic syndrome. It was found that plasma free protein S was reduced in three out of the five patients studied. Thus, acquired free protein S deficiency does occur in children with nephrotic syndrome and is one of many factors which may place them at risk for a thromboembolic event. PMID- 8726223 TI - Severe combined immunodeficiency due to a defect in the tyrosine kinase ZAP-70. PMID- 8726224 TI - The glucocorticoid-glucocorticoid receptor signal transduction pathway, transforming growth factor-beta, and embryonic mouse lung development in vivo. AB - Lung morphogenesis has been shown to be regulated by glucocorticoids (CORT). Because CORT has been primarily thought to affect fetal lung development, previous studies have focused on the role of CORT receptor (GR)-mediated regulation of fetal lung development. Although endogenous CORT increases during embryonic and fetal stages and exogenous CORT treatment in vivo and in vitro clearly accelerates embryonic lung development, little is known about the morphoregulatory role of the embryonic CORT-GR signal transduction pathway during lung development. In this study, we characterize the embryonic mouse CORT-GR pathway and demonstrate: stage-specific in situ patterns of GR immunolocalization; similarity in GR relative mobility with progressive (E13 --> E17) development; that embryonic GR can be activated to bind a GR response element (GRE); significantly increasing levels of functional GR with increasing lung maturation; and the presence of heat shock protein (hsp) 70 and hsp90 from early (E13) to late (E17) developmental stages. These results support the purported importance of the embryonic CORT-GR signal transduction pathway in progressive lung differentiation. To demonstrate that the embryonic CORT-GR directed pathway plays a role in lung development, early embryonic (E12) lungs were exposed to CORT in utero and surfactant-associated protein A (SP-A) expression was analyzed; CORT treatment up-regulates SP-A mRNA expression and spatiotemporal protein distribution. Finally, to determine whether CORT-GR directed pulmonary morphogenesis in vivo involves the modulation of growth factors, we studied the effect of CORT on TGF-beta gene expression. Northern analysis of TGF-beta 1, TGF-beta 2, and TGF-beta 3 transcript levels in vivo indicates that CORT regulates the rate of lung morpho- and histodifferentiation by down-regulating TGF-beta 3 gene expression. PMID- 8726226 TI - Glucose is arrhythmogenic in the anoxic-reoxygenated embryonic chick heart. AB - Unlike in adult heart, embryonic myocardium works at low PO2 and depends preferentially on glucose. Therefore, activity of the embryonic heart during anoxia and reoxygenation should be particularly affected by changes in glucose availability. Hearts excised from 4-d-old chick embryos were submitted in vitro to strictly controlled anoxia-reoxygenation transitions at glucose concentrations varying from 0 to 20 mmol/L. Spontaneous and regular heart contractions were detected optically as movements of the ventricle wall and instantaneous heart rate, amplitude of contraction, and velocities of contraction and relaxation were determined. Anoxia induced transient tachycardia and rapidly depressed contractile activity, whereas reoxygenation provoked a temporary and complete cardioplegia (oxygen paradox). In the presence of glucose, atrial rhythm became irregular during anoxia and chaotic-periodic during reoxygenation. The incidence of these arrhythmias depended on duration of anoxia, and no ventricular ectopic beats were observed. Removal of glucose or blockade of glycolysis suppressed arrhythmias. These results show similarities but also differences with respect to the adult heart. Indeed, glucose 1) delayed and anoxic contractile failure, shortened the reoxygenation-induced cardiac arrest, and improved the recovery of contractile activity; 2) attenuated stunning at 20 mmol/L but worsened it at 8 mmol/L; and 3) paradoxically, was arrhythmogenic during anoxia and reoxygenation, especially when present at the physiologic concentration of 8 mmol/L. The last named phenomenon seems to be characteristic of the young embryonic heart, and our findings underscore that fluctuations of glycolytic activity may play a role in the reactivity of the embryonic myocardium to anoxiareoxygenation transitions. PMID- 8726225 TI - Deficiency of the voltage-dependent anion channel: a novel cause of mitochondriopathy. AB - A patient with a deficient voltage-dependent anion channel (VDAC) is reported, presenting clinically with psychomotor retardation and minor dysmorphic features. Biochemical studies on muscle mitochondria showed impaired rates of pyruvate oxidation and ATP production; however, no specific deficient activity of one of the mitochondrial enzymes was involved. Western blotting experiments indicated an almost complete VDAC deficiency in skeletal muscle. The only moderately decreased VDAC content in the patient's fibroblasts might indicate that VDAC is expressed in a tissue-specific manner. The deficiency is likely caused by a mutation in the HVDAC1 gene or by a distributed posttranslational modification. This is the first described deficiency of a component of the outer mitochondrial membrane associated with the pyruvate oxidation pathway. Defects in this membrane should be considered as a possible cause of otherwise unexplained mitochondrial disorders. PMID- 8726227 TI - Sympathetic activity in children undergoing balloon valvuloplasty of pulmonary stenosis. AB - We studied the influence of balloon valvuloplasty on alpha- and beta-adrenoceptor densities, plasma catecholamine, and cAMP levels in children and infants with pulmonary stenosis before and 10 min after balloon dilatation, employing as controls children undergoing transcatheter occlusion of patent ductus arteriosus (PDA) with Qp/Qs ratio < 1.5. In the PDA group, the alpha-adrenoceptor density (Bmax) was 3.75 +/- 0.72 fmol/10(7) cells (n = 15) before occlusion and remained unchanged at 3.35 +/- 0.47 fmol 10 min thereafter. In the pulmonary stenosis patients (n = 31), the receptor density was 59% higher (p < 0.05) before, and decreased to PDA levels 10 min after, the procedure. The control beta adrenoceptor density was 64.8 +/- 11.0 fmol/10(6) cells before, and 71.2 +/- 13.2 fmol 10 min after, occlusion. In the study group, the density was 23% lower (p < 0.07) and increased to the PDA levels 10 min after the dilatation. Compared with the PDA, pre- and postdilatation plasma norepinephrine levels were not significantly changed; epinephrine was slightly elevated before, but increased by 73% after, dilatation; dopamine was 80% (p < 0.05); and cAMP was 37% higher before, and remained elevated at 70 and 23% above the PDA values after, the procedure. Accordingly, alpha-adrenoceptor density is significantly elevated in children with pulmonary stenosis and decreases significantly immediately after balloon valvuloplasty. On the other hand, beta-adrenoceptor density is attenuated and increases toward normal levels after the procedure. The immediate reversal of the receptor levels after balloon valvuloplasty suggests that this procedure exerts acute effects on the sympathetic functional level in this disease. PMID- 8726228 TI - The ontogeny of cerebral and cerebellar nitric oxide synthase in the guinea pig and rat. AB - The appearance of nitric oxide synthase (NOS, EC 1.14.13.39) activity in the brain of fetal and neonatal guinea pigs and rats was studied. In the guinea pig, NOS increased from an almost undetectable level at 0.49 of gestation (31 d), reaching adult levels before birth and peaking at 140% of the adult activity (forebrain) or 250% of the adult activity (cerebellum) in the week after birth. The rise in fetal NOS activity followed the reported rise in the estrogen receptor concentration in the brain and could be reduced by treatment of the guinea pig at full term with tamoxifen, implicating estrogens in the expression of fetal NOS activity. In the rat, brain NOS activity did not rise significantly until after birth, reaching adult levels approximately 2 wk after birth, and rising to 150 or 130% of the adult activity in the forebrain and cerebellum, respectively, at 4 wk after birth. The appearance of NOS activity in the rat also followed the reported appearance of estrogen receptors in the brain. In both species the appearance of high NOS activity in the brain immediately precedes the period in which maximal synaptogenesis occurs: immediately before birth in the guinea pig and 2-3 wk after birth in the rat. Thus the appearance of a functional estrogen-estrogen receptor system in the brain may be responsible, at least in part, for the expression of a high activity of NOS, which in turn may play important roles in promoting cerebral blood flow and synaptogenesis in the developing brain. PMID- 8726229 TI - Inducible nitric oxide synthase expression in cerebrovascular smooth muscle and neutrophils after traumatic brain injury in immature rats. AB - The inflammatory response after traumatic brain injury (TBI) includes cytokine production, leukocyte infiltration, and microglial activation. Production of nitric oxide by inducible nitric oxide synthase (iNOS) occurs during acute inflammation outside of the CNS and in models of cerebral ischemia, and therefore may contribute to the inflammatory response after TBI. The purpose of this study was to localize and define the time course of iNOS expression after TBI in the immature rat. Immature Wistar rats (age 3.5-4.5 wk) were anesthetized and subjected to percussive trauma to the right parietal cortex. Nontraumatized rats were used as controls (n = 7). At 2, 24, 48, or 168 h (n = 3/group) posttrauma rats were killed by perfusion fixation. Brains were removed, frozen, sectioned, immunostained with antibodies against iNOS and glial fibrillary acidic protein (GFAP, a marker specific for astrocytes), and imaged using fluorescent detection systems. There was no detectable expression of iNOS in control brains. At 2h, minimal cerebrovascular iNOS expression was seen in the peritrauma area. At 24 and 48 h, there was marked peritrauma cerebrovascular iNOS expression that appeared to be restricted to vascular smooth muscle cells and infiltrated leukocytes. Further dual-immunolabeling showed that the leukocytes expressing iNOS were predominantly neutrophils. At 168 h, iNOS expression was no longer detectable. iNOS was not detectable in GFAP-positive cells. The prominent expression of iNOS protein after TBI in cerebrovascular smooth muscle cells and infiltrated neutrophils suggests that iNOS may play a role in cerebrovascular disturbances and secondary brain injury after trauma. PMID- 8726230 TI - Accumulation of cytotoxins during the development of seizures and edema after hypoxic-ischemic injury in late gestation fetal sheep. AB - Several hours after an hypoxic-ischemic injury to the developing brain, hyperemia, then seizures, edema, and infarction can develop. The roles of nitric oxide (NO) synthesis and excitotoxin accumulation during these later phases of injury are not known. The time course of extracellular levels of amino acids within the parasagittal parietal cortex were measured with microdialysis during and for 3 d after 30 min of cerebral ischemia in nine chronically instrumented near-term fetal sheep (119-133 d). Cortical electroencephalographic (EEG) activity and extracellular space (ECS) were quantified simultaneously with real time spectral analysis and cortical impedance measurements, respectively. Amino acid concentrations were measured using HPLC. During ischemia, citrulline (by product of NO synthesis), glutamate, glycine, and gamma-aminobutyric acid (GABA) concentrations rose to 147 +/- 18%, 180 +/- 20%, 290 +/- 50% and 4800 +/- 1300% of baseline respectively (p < 0.05). The excitotoxic index ([glutamate] x [glycine]/[GABA]) decreased to 15 +/- 8%. Upon reperfusion, the cytotoxic edema and amino acid accumulation largely resolved within 1 h, and the EEG was depressed. Citrulline began to rise again by 4 h (p < 0.05), reaching a maximum (273 +/- 21%) at 32 +/- 2 h. Seizure activity developed at 7 +/- 2 h, and impedance plus the excitotoxic index then rose progressively and peaked at 32 +/- 2 h (480 +/- 170%). At 72 h, there was severe neuronal loss and laminar necrosis within the parasagittal cortex. These data suggest that, several hours after a severe hypoxicischemic injury, NO synthesis increased, then seizures arose, and edema developed concomitantly with the accumulation of excitotoxins. PMID- 8726231 TI - Thrombin inhibitors reduce intrapulmonary accumulation of fibrinogen and procoagulant activity of bronchoalveolar lavage fluid during acute lung injury induced by pulmonary overdistention in newborn piglets. AB - We determined whether antithrombin (AT III) or hirudin (a specific thrombin inhibitor) reduce both the accumulation of fibrinogen in lung parenchyma and the procoagulant activity of bronchoalveolar lavage (BAL) fluid during acute lung injury induced by pulmonary overdistention. Newborn piglets were randomized to six-hourly infusions of AT III concentrate, a continuous infusion of recombinant hirudin, or no anticoagulant therapy. All animals were subjected to 24 h of identical mechanical ventilation at high peak pressures (3.9 kPa or 40 cm H2O). Tidal volumes were raised to a mean of 69 mL/kg in all three groups. Mean AT III levels in supplemented piglets (n = 22) were increased to 1.46 (SD 0.24) U/mL at 24 h, compared with 0.67 (SD 0.16) U/mL in controls (n = 23). The median activated partial thromboplastin time in animals receiving hirudin (n = 18) was prolonged to 53 s versus 34 s in untreated animals. The intrapulmonary accumulation of i.v. administered 125I-fibrinogen was reduced by AT III concentrate or hirudin, compared with untreated littermates (p = 0.003). The procoagulant activity of BAL fluid was also decreased by both thrombin inhibitors (p = 0.001). Intrapulmonary accumulation of fibrinogen and the procoagulant activity of BAL fluid were reduced by AT III or hirudin during lung injury caused by pulmonary overdistention. Future investigations should determine whether tangible clinical benefits result from this reduced potential for fibrin deposition in the injured lung. PMID- 8726232 TI - Content of dipalmitoyl phosphatidylcholine in lung surfactant: ramifications for surface activity. AB - The content of dipalmitoyl phophatidylcholine (DPPC) in the phosphatidylcholine (PC) fraction of calf lung surfactant extract (CLSE) is measured by gas chromatography (GC) and estimated from the widely used osmium tetroxide assay for disaturated phosphatidylcholine (DSPC). The surface-active properties of model phospholipid/apoprotein surfactants with varying DPPC content are also defined and compared relative to CLSE. GC analysis of fatty acids in PC isolated from CLSE indicated a possible range of 30 to 65% for DPPC content depending on C16:0 fatty chain mismatching, and further studies using phospholipase A2 treatment indicated an actual DPPC content < or = 41%. The osmium tetroxide assay gave a very high value of 70% for the DSPC content of surfactant PC, and experiments with synthetic phospholipids demonstrated that this assay responded inappropriately in the presence of monounsaturated PC, leading to falsely elevated DSPC values. The influence of DPPC content on adsorption and film behavior was investigated in model surfactants containing 40, 60, and 80% DPPC (DPPC/egg PC/egg PG, 40:50:10, 60:30:10, and 80:10:10 by mol) combined with 1.3% hydrophobic surfactant protein (SP)-B and -C. The biophysical properties of the model surfactant with 40% DPPC were found to be closer to CLSE than those of mixtures with 60 or 80% DPPC. The adsorption of dispersions containing 40% DPPC with 1.3% SP-B, C was almost identical to CLSE and was improved in rate and magnitude compared with the mixtures with higher DPPC content (60 or 80%). In Wilhelmy balance studies of cycled films, respreading was increased and maximum surface pressure was decreased for the 40% versus higher DPPC content mixtures, again approaching CLSE in behavior. All synthetic phospholipid (SPL):SP mixtures lowered surface tension to < 1 mN/m in oscillating bubble studies at physiologic cycling rate (20 cpm), but the 40% DPPC mixture had a time dependent most closely matching that of CLSE. Our measured DPPC content near 40% for lung surfactant PC, and the similarly high activity of a related synthetic phospholipid/apoprotein model mixture, suggest that exogenous surfactants with relatively low DPPC contents might be important for future study and development. PMID- 8726233 TI - Correction by gene expression of biochemical abnormalities in fibroblasts from Zellweger patients. AB - Zellweger syndrome is a prototype of peroxisomal biogenesis disorders and a fatal autosomal recessive disease with no effective therapy. We identified nine genetic complementation groups of these disorders, and mutations in peroxisome assembly factor-1 (PAF-1) and the 70-kD peroxisomal membrane protein (PMP70) genes have been detected by our group F and Roscher's group 1, respectively. We now describe permanent recovery from generalized peroxisomal abnormalities in fibroblasts of a Zellweger patient from group F, such as biochemical defects of peroxisomal beta oxidation, plasmalogen biosynthesis, and morphologic absence of peroxisomes, by stable transfection of human cDNA encoding PAF-1. In the light of these observations, we designed a gene expression system using fibroblasts from patients with peroxisomal biogenesis disorders. In Zellweger fibroblasts obtained from Roscher's group 1 and transfected with human cDNA encoding PMP70, peroxisomes were not morphologically identifiable, and peroxisomal function did not normalize. PMID- 8726234 TI - First trimester prenatal diagnosis of Smith-Lemli-Opitz syndrome (7 dehydrocholesterol reductase deficiency). AB - In Smith-Lemli-Opitz syndrome (SLOs), 7-dehydrocholesterol (7-DHC) accumulated because there is a block in the pathway for synthesis of cholesterol via 7-DHC. Prenatal diagnosis of SLOs has been achieved by analysis of 7-DHC in amniotic fluid obtained at 16-18 wk from pregnancies at risk. The purpose of this study was to investigate 7-DHC and cholesterol concentrations in chorionic villus (CV) samples with a view to performing first trimester prenatal diagnosis. Using a sensitive gas chromatography-mass spectrometry assay it was possible to detect 7 DHC in CV samples obtained as early as 7 wk of gestation. The ration of 7-DHC to cholesterol in placental tissue was shown to be relatively constant over the gestational period of 7-18 wk. We therefore proceeded to analyze the 7 DHC/cholesterol ration in CV samples taken at 10-12 wk of gestation from three pregnancies at risk for SLOs. The results were as follows: patient A, 1.10 x 10( 3); patient B, 1.80 x 10(-3); patient C, 0.091; control range for CVS (8-12 wk), 3.10 x 10(-4) to 1.62 x 10(-3) (mean +/- 2SD; n = 5). The fetus of patient C was diagnosed as affected by SLOs, and the parents requested termination. Analysis of cultured skin fibroblasts confirmed the diagnosis. Pregnancies A and B were diagnosed unaffected, and this was confirmed first by amniocentesis and then by the birth of normal infants at term. We conclude that synthesis of cholesterol via 7-DHC is occurring in the placenta and/or fetus at 10 wk of gestation and that prenatal diagnosis by CV biopsy is possible. PMID- 8726235 TI - The highest concentration of primitive hematopoietic progenitor cells in cord blood is found in extremely premature infants. AB - We used two independent in vitro assays to measure the frequency and proliferative potential of primitive hematopoietic progenitors from the cord blood of 23-41 wk of gestation newborns and adult bone marrow. The frequency of primitive progenitors in the circulating blood cells of infants at 23-31 wk of gestation was significantly greater than the frequency in adult bone marrow or cord blood of more mature newborns. In addition, on a cell to cell basis, the proliferative potential of the primitive progenitors form immature infants (23-31 wk) was greater than in adult bone marrow or cord blood of term newborns. Circulating cord blood cells from immature infants were used as targets for transduction with recombinant retrovirus vectors, and a high efficiency of gene transfer was observed in both primitive and committed progenitors. These data demonstrate that there are major ontogenic shifts in primitive progenitor/stem cell populations in the circulation throughout development as well as programmatic changes in hematopoietic progenitor cell proliferation. In addition, fetal cord blood cells may prove useful targets for genetic manipulation and autologous transplantation. PMID- 8726236 TI - Decreased thrombin activity of fibrin clots prepared in cord plasma compared with adult plasma. AB - We hypothesized that the immaturity of the newborn coagulation system may influence the procoagulant activity of clotbound thrombin. 125I-Labeled fibrin clots were prepared from adult and cord plasma, incubated in their respective plasmas, and fibrinopeptide A (FPA) production was measured. Cord plasma clots generated significantly less FPA compared with adult plasma clots (p < 0.001). Cord plasma clots incubated in adult plasma generated similar amounts of FPA as cord plasma clots in cord plasma. Adult plasma clots incubated in cord plasma clots generated more FPA than adult plasma clots in adult plasma. Adult and cord plasma clots were then incubated with purified human adult fibrinogen, and the discrepancy between adult and newborn plasma clots remained (p < 0.01). To compare the amount of clot bound thrombin, adult and cord plasma clots were sonicated and incubated with fibrinogen. Again, significantly less thrombin was seen in cord clots compared with adult clots (p < 0.01). Because cord plasma has lower prothrombin concentrations (0.5 U/mL) we increased cord plasma prothrombin concentration by the addition of purified prothrombin. Prothrombin supplemented cord plasma clots generated more thrombin than unsupplemented clots (p < 0.01) and in amounts similar to the adult system. In conclusion, decreased amounts of thrombin present in cord plasma clots compared with adult plasma clots results in less FPA production. The low plasma concentration of prothrombin in cord plasma is responsible for this phenomenon. PMID- 8726238 TI - Granulocyte functions in children with cancer are differentially sensitive to the toxic effect of chemotherapy. AB - To analyze the toxicity associated to chemotherapy upon granulocytes, different functional assays were performed, within days of drug exposure and at time of bone marrow recovery, on polymorphonuclear neutrophils (PMN) from children with cancer. There were no significant postchemotherapy changes in the expression of the different receptors studied nor in the phagocytosis of Staphylococcus aureus 42D. By contrast, a significant decrease was observed in H2O2 production in PMN recently exposed to chemotherapy with both cytofluorometric and chemiluminescence assays. There was also a decrease in the production of O2- and in chemotaxis; finally, the intracellular killing of S. aureus 42D and Escherichia coli was reduced. In patients having recovered from drug-induced bone marrow aplasia, PMN functions were found to be normal except for bactericidal activity which was still defective. The observations indicate that, in patients exposed to chemotherapy, some PMN functions are transiently altered, whereas microorganism cell killing is continuously impaired. PMID- 8726237 TI - Management of late anemia in Rhesus hemolytic disease: use of recombinant human erythropoietin (a pilot study). AB - The management of (Rhesus) hemolytic disease of the fetus and newborn includes intrauterine transfusions to prevent the development of hydrops, treatment of the possible hyperbilirubinemia in the immediate postnatal period, and treatment of late anemia. Low levels of serum erythropoietin due to suppression of the bone marrow by multiple intrauterine transfusions is a suggested mechanism for this anemia. The aim of our study was to test whether recombinant human erythropoietin reduced the need for erythrocyte transfusions in these infants. Twenty infants with Rhesus isoimmunization were blindly randomized to treatment and control groups at the 2nd wk of life. The number of intrauterine and exchange transfusions and demographic data were similar in both groups. The infants in the treatment group received recombinant human erythropoietin, s.c. 200 U/kg of body weight three times a week for a period of 6 wk, whereas the infants in the control group received a placebo for the same period. In the treatment group, the mean number of erythrocyte transfusions was significantly lower than that of the control group (1.8 versus 4.2). The reticulocyte counts and Hb levels rose earlier in the treatment group. The platelet and neutrophil counts were similar in both groups throughout the study. This study demonstrates that recombinant human erythropoietin treatment decreases the need for erythrocyte transfusions in the late anemia of infants with Rh isoimmunization. Considering the risks of blood transfusions, this decrease in the donor exposure is worthwhile. PMID- 8726239 TI - Neutrophil respiratory burst in term and preterm neonates without signs of infection and in those with increased levels of C-reactive protein. AB - Developmental immaturities in neonatal host defense predispose the neonates to an increased mortality rate during bacterial infections. Early diagnosis is of great clinical importance, but, especially in neonates, is sometimes very difficult. The ability to generate reactive oxygen species, the so-called respiratory burst, is essential for neutrophils to kill infectious microorganisms. Therefore, changes of respiratory burst may reflect increased susceptibility of neonates to infections and may be useful for the early detection of infections. Superoxide anion production was determined by a flow cytometric method using dihydrorhodamine 123 (DHR) as an oxidative probe after priming of neutrophils with PBS buffer (spontaneous burst), with N-formyl-methionyl-leucyl-phenylalanine (fMLP), or with Escherichia coli. During the study period, the spontaneous percentage of activated cells in whole blood as well as the percentage of activated cells in stimulation with fMLP was lower in adults (n = 100; PBS, 1.0 +/- 0.1%; fMLP, 8.3 +/- 0.9%) compared with neonates without signs of infection (n = 143). Among the latter, the percentage of activated cells (PBS and fMLP assay) varied with respect to gestational age and hours of life: lowest values were measured in preterm newborns with gestational age less than 32 wk and between 25 and 120 h of life. The same correlation to gestational age was true for total neutrophil cell counts. In neonates with increased levels of C-reactive protein during the first 5 d of life (n = 43), the percentages of activated cells after PBS and fMLP incubation were higher than those of neonates without signs of infection. The relationship of neutrophil respiratory burst and neutrophil cell counts to gestational age might reflect at least in part a reason for the increased susceptibility of neonates to infections. Furthermore, determination of respiratory burst may prove to be a new laboratory parameter of neonatal infection. PMID- 8726240 TI - Insulin-like growth factor binding protein-3 generation: an index of growth hormone insensitivity. AB - GH insensitivity may be an inherited condition or may arise as a consequence of disease of malnutrition. Laron syndrome is the most severe form of GH insensitivity, arising from an absent or defective GH receptor. Less severe forms of GH insensitivity, however, may exist, resulting in short stature but in few other features of Laron syndrome. We have identified a heterogeneous group of children with short stature and either high basal (> 10 mU/L) or high peak GH levels (> 40 mU/L) on GH provocation testing, to examine biochemical markers of GH sensitivity. These children received 4 d of GH (0.1 U/kg) and the increment in IGF-I, IGF binding protein (BP)-3, and GHBP was determined. Eight GHD children, commencing GH therapy, were recruited as positive controls. The two groups could not be differentiated by age, height SDS (SD score), height velocity SDS, or body mass index. IGF-I and IGFBP-3 generation were correlated in all children (delta SDS IGF-I versus delta SDS IGFBP-3, r = 0.49, p = 0.03). Neither basal GHBP levels or the increment in GHBP were predictive of the IGF-I or IGFBP-3 response to GH. The GHI group had a significantly reduced IGFBP-3 response to stimulation with 4 d of GH (median percent increment in IGFBP-3, 26%, versus 72% in the GHD group, P = 0.03); their IGF-I response to GH was also reduced (median % increment in IGF-I 75% versus 144% in the GH deficient group), but this did not achieve significance, p = 0.06. In all children, the percentage rise or delta SDS in both IGF-I and IGFBP-3 inversely correlated with the GH peak obtained on provocation testing, the latter being the most significant determinant of GH peak. We propose that the "IGF generation test", in particular IGFBP-3 generation, can be used in the investigation of partial GH insensitivity. Further work, however, is required to establish diagnostic criteria for partial GH insensitivity. PMID- 8726241 TI - Is there a no-effect dose for corticosteroid-induced cleft palate? The contribution of endogenous corticosterone to the incidence of cleft palate in mice. AB - Teratology and genetic counselors are frequently asked whether very low exposures of drugs and chemicals can cause a child's congenital malformations. One critical factor on which the counseling is based is the dose. Because teratogenic effects follow a toxicologic dose-response curve with a no-effect dose, frequently counselors can refute a causal relationship because the dose was far below the no observable-effect dose. Recently, some investigators have suggested that some teratogens which are present in physiologic levels such as cortisone, glucose, insulin, or sex steroids may contribute to the background incidence of congenital malformations and, therefore, there is no safe dose. Using corticosteroid-induced cleft palate in mice as the model, we conducted experiments to test this hypothesis. Adrenalectomy of A/J or CD-1 dams resulted in a reduction of endogenous corticosterone, but did not reduce the spontaneous incidence of cleft palate in the offspring. In A/J mice, the incidence of isolated cleft palate increased with adrenalectomy indicating that the spontaneous incidence of this defect is not due to endogenous corticosterone. Adrenalectomy did not affect the susceptibility of CD-1 mice to cortisone induced cleft palate demonstrating that endogenous corticosterone did not contribute significantly to the incidence of cleft palate induced by the exogenous corticosteroid. Finally, results in CD-1 mice clearly indicate that cortisone, like other teratogens, has a no-effect level for teratogenesis. These studies support the concept of a threshold in the dose-response relationship for corticosteroid-induced cleft palate in mice. PMID- 8726242 TI - Food antigen activates intraepithelial and lamina propria lymphocytes in food sensitive enteropathy in mice. AB - Morphologic and immunologic changes in the gut mucosa of food-hypersensitive mice, from a study model generated by feeding ovalbumin (OVA) to female BALB/c mice after intraperitoneal injection of cyclophosphamide (CY), were investigated in an effort to clarify the mechanisms of food-sensitive enteropathy. Villous atrophy, crypt hyperplasia, and increased numbers of intraepithelial lymphocytes (IEL) were confirmed in the antigen-challenged OVA-sensitive mice as seen in food sensitive enteropathy in humans, whereas no significant morphologic changes were observed in the nontreated control group or groups treated with OVA or CY alone. IEL and lamina propria lymphocytes (LPL) were isolated from the intestinal mucosa before and after the antigen challenge, and surface markers were analyzed by FACScan. After the antigen challenge, the numbers of CD8+ cells increased among the IEL, and the occurrence of both CD4+ and CD8+ cells increased among the LPL. The numbers of Thy-1+ cells and TCR- alpha/beta + cells increased among both the IEL and LPL, and LFA-1 expression was enhanced in both of these lymphocyte populations. The proliferative response of IEL and LPL to OVA increased in a dose dependent manner after the antigen challenge in the OVA-sensitive mouse model. These results indicate that IEL and LPL, possibly those that have migrated from peripheral blood, are activated by orally administered antigens and cause mucosal damage in the food-sensitive enteropathy. PMID- 8726243 TI - Short-term efficacy and safety of pravastatin in 72 children with familial hypercholesterolemia. AB - The safety, tolerability, and efficacy of a 12-wk treatment with pravastatin, 5, 10, and 20 mg/d, was evaluated in 72 children with heterozygous familial hypercholesterolemia (FH) in a double-blind, randomized and placebo-controlled study. The results show that pravastatin was well tolerated and that adverse events were mild and equally distributed among the three treatment groups. Plasma total and LDL cholesterol levels were significantly reduced in all pravastatin treatment groups, in comparison with the control group; -24.6% (-28.1 to 21.0) and -32.9% (-37.0 to -28.6), for mean change and 95% confidence interval, respectively. In four children plasma LDL cholesterol levels were reduced within normal limits for sex and age. HDL cholesterol increased in the pravastatin 20-mg group, +10.8% (+3.4 to +18.8), whereas plasma apo B100 and very LDL (VLDL) cholesterol levels were reduced within all pravastatin-treated groups -26.8% ( 31.2 [corrected] to -21.7) and -24.5% (-35.0 to -12.3). These data show that short-term pravastatin treatment of children with FH is safe and effective, although long-term dose titration studies with 3-hydroxy-3-methylglutaryl-CoA reductase inhibitors need to be performed, to reduce plasma LDL cholesterol levels below a predefined level. The results of these studies have to be awaited before new treatment strategies are to be considered in these children. PMID- 8726244 TI - Effect of resistant and digestible starch on intestinal absorption of calcium, iron, and zinc in infant pigs. AB - The first nonmilk foods that are given to infants contain high levels of starch, a fraction of which is resistant to enzyme hydrolysis. Incomplete digestion of starch may interfere with the absorption of certain minerals. A fraction of dietary starch which is resistant to in vitro enzymatic hydrolysis has been termed resistant starch. The aim of this study was to compare the intestinal apparent absorption of calcium, phosphorus, iron, and zinc in the presence of either resistant or digestible starch. Twelve 7-10-d-old piglets were fitted with a T-tube inserted into the intestine approximately 3 m distal to the duodenum. Animals received in random order 200 mL of a test meal of cooked, cooled, high amylose corn starch (16.4% resistant starch), or cooked rice starch (digestible starch) administered by an orogastric tube. Both meals contained the same amount of calcium, phosphorus, iron, and zinc. The test meal also contained tracer amounts of 59Fe and 65Zn, as well as polyethylene glycol 3350, as a nonabsorbable marker. Intestinal apparent absorption of starch was greater the meal with digestible starch (71.0 +/- 17.0%) than after the meal with resistant starch (49.2 +/- 10.3) (p < 0.001). After feeding the meals with resistant and digestible starch, mineral apparent absorption was, respectively: calcium, 40.2 +/- 11.8% versus 28.1 +/- 16.4% (p < 0.05); phosphorus, 73.2 +/- 14.0% versus 67.8 +/- 18% (NS); iron, 24.1 +/- 12.2% versus 12.6 +/- 10.6% (p < 0.01), and zinc, 35.0 +/- 13.0% versus 30.6 +/- 8.22% (NS). In conclusion, a meal containing 16.4% resistant starch resulted in a greater apparent absorption of calcium and iron compared with a completely digestible starch meal. If this finding holds true for the whole bowel, administration of resistant starches could have a positive effect on intestinal calcium and iron absorption. PMID- 8726245 TI - Heterogeneity of intestinal lactase activity in children: relationship to lactase phlorizin hydrolase messenger RNA abundance. AB - Despite extensive study in both humans and nonhuman mammals the mechanisms which regulate intestinal lactase activity, particularly during development, are incompletely understood. Our previous studies of human adults are consistent with an important role of lactase-phlorizin hydrolase (LPH) mRNA abundance in determining the lactase persistence/nonpersistence phenotypes. Our intent in the present study was to determine the role of LPH mRNA in the regulation of lactase in children. We therefore studied duodenal mucosal biopsies from 39 children undergoing diagnostic upper endoscopy in whom significant small intestinal and nutritional disease was excluded. We found no relationship between the level of LPH mRNA and lactase enzymatic activity. Our observations suggest the importance of posttranscriptional mechanisms in lactase regulation in human children. PMID- 8726246 TI - Visual acuity and fatty acid status of term infants fed human milk and formulas with and without docosahexaenoate and arachidonate from egg yolk lecithin. AB - Preterm infants fed formulas with docosahexaenoic acid (DHA, 22:6n-3) during the interval equivalent to the last intrauterine trimester and beyond have higher circulating DHA and transiently higher visual acuity compared with infants fed formulas containing linolenic acid. In term infants several nonrandomized studies of infants receiving DHA from human milk suggest a relationship between DHA status and acuity, but the evidence for a cause-and-effect relationship is mixed. In the present study, term infants were randomly assigned to a standard term formula (n = 20) or the same formula with egg yolk lecithin to provide DHA (0.1%) and arachidonic acid (AA, 20:4n-6, 0.43%) (n = 19) at levels reported in milk of American women. A third group of infants was breast fed for > or = 3 mo (n = 19). Grating visual acuity (Teller Acuity Card procedure) and plasma and red blood cell (RBC) phosphatidylcholine (PC) and phosphatidylethanolamine (PE) DHA and AA were determined at corrected ages of 2, 4, 6, 9 (acuity only), and 12 mo past term = 40 wk postmenstrual age (PMA). At 2 mo breast-fed infants and infants fed the supplemented formula had higher grating acuity than term infants fed standard formula. As in preterm infants, the increase was transient. Plasma PC DHA and AA and RBC PE AA increased by 2 mo in supplemented infants, but RBC PE DHA in supplemented infants was not higher than in controls until 4 mo and beyond. Despite normal intrauterine accumulation of DHA and AA, infants fed formula with 2% linolenic acid and 0.1% DHA had better 2-mo visual acuity than infants fed formula with 2% linolenic acid. PMID- 8726247 TI - Measurement of cranial optical path length as a function of age using phase resolved near infrared spectroscopy. AB - Near infrared spectroscopy (NIRS) has been used to measure concentration changes of cerebral hemoglobin and cytochrome in neonates, children, and adults, to study cerebral oxygenation and hemodynamics. To derive quantitative concentration changes from measurements of light attenuation, the optical path length must be known. This is obtained by multiplying the source/ detector separation by a laboratory measured differential path length factor (DPF) which accounts for the increased distance traveled by light due to scattering. DPF has been measured by time of flight techniques on small populations of adults and postmortem infants. The values for adults are greater than those for newborns, and it is not clear how to interpolate the present data for studies on children. Recent developments in instrumentation using phase resolved spectroscopy techniques have produced a bedside unit which can measure optical path length on any subject. We have developed an intensity modulated optical spectrometer which measures path length at four wavelengths. Two hundred and eighty three subjects from 1 d of age to 50 y were studied. Measurements were made at a fixed frequency of 200 MHz and a source detector separation of 4.5 cm. Results suggest a slowly varying age dependence of DPF, following the relation DPF690 = 5.38 + 0.049A0.877, DPF744 = 5.11 + 0.106A0.723, DPF807 = 4.99 + 0.067A0.814, and DPF832 = 4.67 + 0.062A0.819, where DPF690 is the DPF measured at 690 nm and A is age is expressed in years from full term. There was a wide scatter of values, however, implying that ideally DPF should be measured at the time of each study. PMID- 8726248 TI - Structural and neurobehavioral delay in postnatal brain development of preterm infants. AB - Postnatal brain development of healthy prematurely born infants was assessed to study possible influence of premature birth and early extrauterine environment on structural, biochemical, and functional brain development. Myelination and differentiation of gray and white matter were studied by in vivo magnetic resonance (MR) imaging (MRI), changes in cerebral metabolism by 1HMR spectroscopy (MRS), and changes in early human neurobehavior by the assessment of preterm infant's behavior (APIB). The stage of intrauterine and extrauterine brain development in prematurely born infants at term was compared with the stage of mainly intrauterine brain development in a group of full-term infants. Eighteen preterm infants unremarkable with respect to neurologic and medical status were studied at approximately 2 wk of postnatal age [gestational age (GA) 1: 32.5 +/- 1.2 wk] and again at term (GA 2: 40.0 +/- 1.1 wk). For comparison a group of 13 full-term born infants (GA T: 40.6 +/- 2.1 wk) were studied by MR and six by APIB. When GA 2 to GA 1 was compared, significant maturational changes were found with MRI in gray and white matter and myelination, with 1H MRS in the concentration of N-acetylaspartate and with all scores of APIB. In preterm infants at term (GA 2) compared with full-term infants (GA T) significantly less gray and white matter differentiation and myelination was observed as well as significantly poorer performance in four neurobehavioral parameters (autonomic reactivity, motoric reactivity, state organization, attentional availability). We conclude that MRI and 1H MRS can be used to study postnatal brain development in preterm infants. Structural and biochemical maturation is accompanied by functional maturation as shown with the neurobehavior assessment. Preterm infants at term compared with full-term infants show a structural as well as a functional delay in brain development assessed at 40 wk of postconceptional age. PMID- 8726249 TI - 1995 Public Policy Plenary Symposium: "the crisis in clinical research". PMID- 8726250 TI - Clinical heterogeneity associated with the mitochondrial DNA T8993C point mutation. AB - The mitochondrial DNA (mtDNA) point mutation T8993G has been associated with maternally inherited Leigh syndrome (MILS) when very abundant (> 95%). MILS patients are usually severely affected and die in early infancy. In 1993, a novel T8993C point mutation was described in a juvenile form of Leigh syndrome (LS) characterized by a less aggressive clinical course. We describe four unrelated T8993C patients who had diverse, relatively mild, clinical manifestations. Polymerase chain reaction-restriction fragment length polymphorphism analysis showed that the heteroplasmic T8993C point mutation was very abundant in several tissues from all four patients (94.2 +/- 1.5%) but was less copious in blood from 20 maternal relatives. ATP production in mitochondria isolated from skin fibroblasts in three patients was normal, whereas in one patient it was decreased to 20-35% of controls. These findings suggest that the T8993C mutation is less severe than the more common T8993G mutation. PMID- 8726251 TI - Validation and verification. PMID- 8726252 TI - Quantitative microbiology in the management of burn patients. I. Correlation between quantitative and qualitative burn wound biopsy culture and surface alginate swab culture. AB - The diagnosis and treatment of burn wound infection is commonly determined by clinical impression and the qualitative results of surface swabs. It has been suggested that quantitative bacteriology from burn wound biopsies confirms burn wound infection and improves patient management. Methods for quantitating surface flora have been described, but comparisons with biopsy specimens have been contradictory. The quantitative and qualitative results of 141 pairs of biopsies and surface swabs, from 74 burn patients, were compared. Staph. aureus was the commonest organism isolated (29 per cent of biopsies and 35 per cent of swabs). Recovery of the same set of species from biopsy and swab occurred in 54 per cent of pairs. There was a significant correlation between the bacterial count obtained by biopsy and by surface swab (P < 0.001), but using various threshold values, the predictive value of the counts obtained by one method to predict the counts obtained by the other was poor. Parallel cultures taken on 18 occasions, showed a significant correlation between bacterial counts obtained from two biopsies or two swabs taken simultaneously (P < 0.002), but there was wide variation in bacterial densities from the same burn wound at the same time. Recovery of the same set of species from both biopsies occurred in 56 per cent of pairs, and from both swabs in 50 per cent of pairs. The use of quantitative microbiology in burns is limited by the unreliability of a single surface swab or biopsy to represent the whole burn wound. PMID- 8726253 TI - Quantitative microbiology in the management of burn patients. II. Relationship between bacterial counts obtained by burn wound biopsy culture and surface alginate swab culture, with clinical outcome following burn surgery and change of dressings. AB - The use of quantitative bacteriology in the burns unit has been thought to be efficient in predicting sepsis or graft loss. To examine the relationship between clinical outcome and bacterial densities on and in the burn wound, 69 biopsy/surface swab pairs were collected from 47 patients on 64 occasions, either immediately prior to excision and grafting, or at routine change of dressings. The mean per cent TBSA burn was 16 (range 1-65). There was a significant correlation between log total bacterial count by biopsy with total white cell count and age (P = 0.028), and a significant negative correlation between total bacterial count by swab with per cent TBSA (P = 0.006). There was no significant difference in bacterial counts between patients judged to be a clinical success or clinical failure (72 h follow-up), either after undergoing excision and grafting, or change of dressings, and no difference in counts between patients with perioperative bacteraemia and those without. With burns > 15 per cent TBSA, a relationship between bacterial counts and subsequent sepsis or graft loss still was not demonstrated. It is suggested that quantitative bacteriology by burn wound biopsy or surface swab does not aid the prediction of sepsis or graft loss. PMID- 8726254 TI - Dispase/detergent treated dermal matrix as a dermal substitute. AB - A method for preparing acellular allogeneic dermal matrix (ADM) and its effectiveness as a dermal substitute are described. Treatment of rat skin with Dispase followed by Triton X-100 completely removed cellular components from the dermis. Subcutaneously implanted ADM evoked no immunological reaction and 20 weeks after implantation, the size of the implanted ADM was reduced to about 60 per cent of its original area. ADM became completely vascularized within 2 weeks after implantation into full thickness skin defects in the rat and inhibited extensive wound contracture. A second layer of ADM placed onto the implanted ADM served as an excellent dressing, providing mechanical protection and permitting vascularization of the underlying implant. Onlay skin autografts placed onto vascularized allogeneic ADM showed good survival when the skin was grafted more than 1 week after ADM implantation. Dispase/detergent treated ADM derived from animal or human skin may be useful in full thickness skin defects providing a vascularized bed for subsequent epidermal coverage. PMID- 8726255 TI - Cultured epidermal sheet grafting with Hemaseel HMN fibrin sealant on nude mice. AB - Grafting of cultured epidermal sheets is a promising technique for skin restoration in extensive burns, but the technique has some limitations, resulting in variable graft takes. These experiments were designed to evaluate the innocuity of Hemaseel HMN fibrin sealant in the grafting process and in vivo evolution of cultured epidermis. A total of 30 mice were grafted, 15 were controls, 15 received tissue sealant application before the deposition of the cultured human epidermal sheets. Seven days after transplantation, compared to controls, the percentage of graft take over the total surface area grafted was greater in animals that had received the tissue sealant application. No difference was found 14 and 21 days postgrafting. In contrast, the percentage of graft take over the bony area (spinal) was significantly increased in animals grafted with previous application of sealant compared to controls at 7, 14 and 21 days postgrafting. Immunohistological and ultrastructural analysis showed that the evolution of the cultured human epidermis after transplantation was similar in both groups. The basement membrane was well structured 21 days after transplantation. The sealant was present at 4 days but not at 21 days postgrafting. Therefore, we conclude that the application of fibrin sealant before cultured epidermal sheet deposition on nude mouse graft bed is innocuous and enhances their mechanical stability. Since in this nude mouse system Hemaseel HMN fibrin sealant increased the percentage of graft take over areas difficult to engraft, we think that it may be advantageous in cultured epidermal sheet grafting on burn patients. PMID- 8726256 TI - Systemic factors influencing the growth of cultured epithelial autograft. AB - This is a retrospective study of 23 consecutive patients who had biopsies taken for cultured epidermal autograft between February 1993 and September 1994. Keratinocyte cultures were initiated on all of these biopsies. Of these 23 biopsies, it was noted that the cells obtained from three patients grew particularly slowly or failed to grow at all. Taking into account that the biopsy and culture conditions were standard for all patients, we investigated whether the patient's previous medical history may have had any effect on the ability of the cells to be cultured in vitro. Our results indicated that the keratinocyte cultures of patients with a significant past medical history, and particularly those with underlying pathology affecting their general physical condition, have a decreased growth rate. This raises the question that general patient condition can possibly influence the clinical use of the technique. PMID- 8726257 TI - Human skin histology as demonstrated by Herovici's stain: a guide for the improvement of dermal substitutes for use with cultured keratinocytes? AB - The potential for the widespread use of cultured keratinocytes for burns treatment is handicapped by practical problems such as fragility, poor take and, often, unsatisfactory cosmesis. Although dermal equivalents reduce these problems there remains a lack of consensus on what is the best structure of such equivalents. At present the commonest support is type I collagen. This histological study, however, using Herovici's stain, clearly shows that in human skin from a variety of anatomical sites the epidermis is not in direct contact with type I collagen but rather with a distinct layer of type III collagen. We suggest that dermal equivalents may have to be constructed so as to include a layer of type III collagen at the interface between the keratinocytes and a type I collagen neo-dermis, so mimicking normal skin structure more closely. PMID- 8726258 TI - The pulsed tuneable dye laser as an aid in the management of postburn scarring. AB - Permanent erythematous scarring following burns is a common problem which may be disfiguring and distressing for the patient. In this case the pulsed tuneable dye laser (585 nm) was used to treat the areas of scarring and also used prophylactically on an active scar. PMID- 8726259 TI - Histological characteristics of the healing process of frozen skin allograft used in the treatment of burns. AB - Combined transplantation of skin autograft and allograft was used for the treatment of severe burns. The allografts were obtained from cadavers and were pretreated with 15 per cent glycerol for 2 h at 4 degrees C then frozen at -80 degrees C until used. Patches of autografts were placed over the burns and were covered by a stretched mesh of allografts. Biopsy samples of transplanted skin were obtained from 5 days to 4 weeks after grafting. Sections were examined by histological and immunohistochemical strainings. At 4 days, the epidermal-dermal junction of allografted skin was separated due to migration of epithelial cells derived from autograft epidermis or from skin appendages of recipient dermis. At 2 weeks, dermal fibroblasts and capillaries proliferated in autografts. At 3 weeks, the dermal components of the allograft were covered by epithelial cells from recipient tissue and were invaded by fibroblasts and capillaries. At 4 weeks, allografted skin was replaced by granulation tissue, which mediated the adhesion of the grafts to the underlying tissue. Skin allografts with a freeze thawing pretreatment provide an appropriate matrix for the epithelial relining and for the growth of granulation tissue in burned skin. PMID- 8726260 TI - Markedly increased lymphatic removal of hyaluronan from skin after major thermal injury. AB - Hyaluronam (HYA) (formerly hyaluronic acid) is an important constituent of the interstitial matrix in skin. Following major burn injury plasma HYA is known to increase markedly. The present study investigated to what extent the lymphatic removal of HYA from skin is affected following major burn injuries. Under ketamine anaesthesia a 30 per cent full thickness scald injury was inflicted on sheep previously provided with a chronic prefemoral lymph fistula. Animals were resuscitated for 24 h using lactated Ringer's infusion as needed to restore and maintain cardiac output within +/- 10 per cent of baseline. Following burn injury, lymph flow from thermally injured tissue gradually increased to 10-25 times above baseline. The baseline HYA concentration in lymph was 4.3 +/- 1.1 micrograms/ml; the HYA concentration in lymph was not significantly altered postinjury. The lymphatic HYA flux (flow x concentration) increased postinjury to 5-30 times control. The HYA concentration in plasma was 162 +/- 12 ng/ml at baseline; postinjury plasma HYA peaked at two to three times baseline at 4-8 h after the injury. At 24 h postinjury plasma HYA was reduced compared to baseline. In conclusion, after major burn injury the lymphatic transport of HYA from the injured area into the systemic circulation increased markedly, leading to elevated plasma concentrations of this high molecular weight connective tissue component. PMID- 8726261 TI - Domestic central heating radiators: a cause for concern in all age groups. AB - A retrospective analysis of all burns admitted to the Welsh Regional Burns and Plastic Surgery Unit, Chepstow, in the period 1 January 1990 to 1 October 1993, highlighted a group of 50 patients who had sustained contact burns from the radiators of domestic central heating systems. There was a male prevalence, with an average age of 43.4 years (range 6 months to 100 years). The mean TBSA burned was 1.58 per cent (range 0.13-6.0 per cent) and half of the injuries were full thickness depth. The forearm and hand were predominantly injured. Although the literature has indicated that the incidence of contact burns peaks at the extremes of the age spectrum, this study has shown that contact radiator burns can be sustained by all age groups. The aim of the audit was to investigate the mechanism of injury and link precipitating factors. The contribution of the high surface temperature of the radiator to the burn injury is alluded to. The various methods available to reduce this risk are discussed and the use of the low surface temperature radiator, already routinely used in health care premises, is advocated. PMID- 8726262 TI - A 1-year prospective study of burns in an Irish paediatric burns unit. AB - Burns are the fourth leading cause of injury death in children in the USA, accounting for 1300 paediatric deaths annually. The majority of paediatric burns mortality and morbidity result from simple domestic accidents that are preventable. A prospective study of paediatric burns from 1 January 1992 to 1 January 1993 was undertaken at our burns unit to outline the profile of the Irish paediatric burns problem. A total of 336 burns were referred to our unit over the 12 months (80 per cent self-referrals, 15 per cent tertiary referrals from district hospitals and 5 per cent GP referrals). Sixteen per cent (57) of the patients required admission and 33 per cent (112) required prolonged dressings as outpatients. Mortality and morbidity rates were comparable to other centres at 1.8 per cent and 39 per cent respectively. The demographic analysis of the patient population was similar to that seen in other studies from developed countries but there were some notable differences. First, there was an alarmingly high incidence of serious sunburn injuries, especially among young infants. Most parents were unaware of the association between childhood sunburn and the development of skin cancer in later life. Second, 90 per cent of the accidents occurred in the home and almost all were preventable. A parent or guardian was present in 87 per cent of cases but parental knowledge of the appropriate first aid measures was poor. It is suggested that a public health education campaign on this issue would help in reducing the incidence and severity of paediatric burn injuries in Ireland. PMID- 8726263 TI - Epidemiology of adults hospitalized with burns in Karachi, Pakistan. AB - Burns are a leading cause of adult death in Karachi slums, therefore we reviewed 1 year's logged experience (November 1992 to October 1993) at Karachi's two adult burn units for patient age, sex, burn severity and outcome. Also 47 inpatients were interviewed regarding their circumstances of injury. We grouped these using Haddon's Matrix. The log identified 832 patients. Females (57 per cent) outnumbered males and were younger on average (25.1 vs 27.6 years, P = 0.002). Females had more severe burns than males (57 per cent vs 50 per cent total body surface area (TBSA) burn, P = 0.002). At the unit with outcome data (n = 556), the case fatality was 56 per cent. The estimated adult mortality due to burns in Karachi was 10.2/100 000, 6.8/100 000 and 14.1/100 000 for men and women, respectively. Burns of interviewed patients were most often associated with flames (33/47), but stove bursts caused the most severe injury (52 per cent TBSA). These patients were predominantly young uneducated female houseworkers, clothed in loose attire who were injured during daylight at home around a floor level stove, unaware of fire safety, and who received no first aid. It was concluded that the high burn severity and case fatality rates demand: (1) preventive measures, such as kitchen sand buckets, safer stove design and placement and education on fire safety and first aid, and (2) risk factor analysis to refine interventions. PMID- 8726265 TI - Transcutaneous metal absorption following chemical burn injury. AB - A 33-year-old metal refinery worker sustained a 40 per cent TBSA burn injury from a mixture containing acids and salts of nickel and cobalt. High levels of both nickel and cobalt were detected in both serum and urine, necessitating treatment with EDTA, a chelating agent. No detrimental effects of either nickel or cobalt have so far been detected, though the patient is being followed in the long term. PMID- 8726264 TI - How big is a hand? AB - Use of the patient's hand to estimate percentage body surface area (BSA) of injury is well established in the management of burns. Exactly what constitutes "the palm of the hand' and how large an area this is, depends on whether you follow Advanced Trauma Life Support teaching. United Kingdom teaching, or use a "Lund and Browder chart'. A study was designed to measure the areas in question to find which was most accurate. The conclusions challenge standard teaching and show a sex difference. The area of the palm alone is 0.5 per cent BSA in males and 0.4 per cent BSA in females, whereas the area of the palm plus the palmar surface of the five digits is 0.8 per cent BSA in males and 0.7 per cent BSA in females. Therefore if a hand alone is used to assess the size of a burn the per cent BSA could be overestimated. PMID- 8726266 TI - Early burn wound excision in "major' burns with "pregnancy': a preliminary report. AB - Burn injury in pregnant patients is not uncommon in developing countries. The results of the management of six pregnant burns patients, admitted during an 18 month period, were analysed. Successful management of burn injuries ranging from 25 to 65 per cent TBSA occurred in patients during the second and third trimester of pregnancy, using early burn wound excision and skin grafting in four patients and by late skin grafting of a granulating wound in one patient. All five patients delivered live babies with no congenital anomalies. One patient with 60 per cent TBSA burns who was unsuitable for early excision, died of septicaemia. This report suggests the need for early burn wound excision and skin grafting in burns patients with pregnancy, in order to improve maternal and fetal survival. However, in developing countries early surgery is not advisable in patients with extensive burns because of the non-availability of biological skin substitutes. PMID- 8726267 TI - Use of patient-controlled analgesia with alfentanil for burns dressing procedures: a preliminary report of five patients. AB - The use of patient-controlled analgesia with alfentanil (PCA-alfentanil) as a form of pain relief for dressing procedures in patients during the acute phase of their burn injuries was investigated. Five ASA 1 and 2 patients with 10-30 per cent total body surface area (TBSA) thermal burns, had PCA-alfentanil for their dressing procedures after standard fluid resuscitation. One patient who did not receive a loading dose and a background infusion of alfentanil had unsatisfactory pain relief. Four patients had good pain relief after a loading dose of IV alfentanil 1 mg followed by a continuous background infusion of 200-800 micrograms/h. Demand dose ranged from 200 to 400 micrograms and lockout time ranged from 1 to 3 min. The total dose of alfentanil delivered ranged from 0.8 to 4.48 mg and duration of the dressings ranged from 30 to 60 min. All patients were mildly sedated, calm, communicative and cooperative during dressing procedures. None of them experienced hypotension or respiratory depression. One patient experienced nausea but no vomiting, no other adverse effects of alfentanil were noted. From the pilot study, PCA-alfentanil may be an effective form of pain relief for dressing procedures in patients during their acute phase of burn injuries. The optimal PCA-alfentanil setting has yet to be determined. PMID- 8726268 TI - Radial free flaps in plantar burns. AB - Problems in the reconstruction of full thickness burns of the sole of the foot are presented. After analysing the existing surgical possibilities for treating this injury, the good results obtained in the forefoot with radial forearm flaps are shown. The main virtues of this flap for treating plantar burns are its suppleness for adaptation to the different contours of the foot. This allows a functional recovery without losing the normal shape of the foot, making the use of prosthetic devices unnecessary. PMID- 8726269 TI - Early, fatal disseminated intravascular coagulation in a patient with 60 per cent burns. AB - A 23-year-old, previously fit and healthy young man was admitted to the West Midlands Regional Burns Unit at the Birmingham Accident Hospital, with 60 per cent body surface area burns and smoke inhalation. On arrival, he was briskly tachycardiac, hypotensive and disorientated. He was admitted to the intensive care unit, intubated and started on intermittent positive pressure ventilation. Despite prompt commencement of resuscitation and the infusion of enormous volumes of colloids, the patient remained oliguric. He soon developed severe haemorrhage from his gastrointestinal, urinary and respiratory tracts. He required several units of blood, but was persistently hypotensive. His condition deteriorated rapidly despite intensive supportive measures. He developed metabolic acidosis, refractory hypotension and died anuric, 20 h later. The post-mortem examination showed the presence of disseminated intravascular coagulation and adult respiratory distress syndrome. PMID- 8726270 TI - Alopecia of the unburned scalp. AB - Alopecia of the unburned scalp, the temporary or permanent loss of hair, is a preventable complication. Patients at highest risk of developing alopecia are those with burn encephalopathy and in a prolonged shock state. The occipital scalp is the most common site and localized pressure-induced ischaemia is the likely cause. Frequent head repositioning and use of devices to decrease the pressure effect may decrease the incidence of this complication. PMID- 8726271 TI - Use of cyanoacrylate glue to repair punctured tissue expanders. PMID- 8726272 TI - Bleeding oesophageal varices. AB - BACKGROUND: The Danish contribution to evaluation and treatment of bleeding oesophageal varices. METHODS: Danish papers dealing with portal hypertension and oesophageal varices have been reviewed and set in relation to international publications. RESULTS: The Danish papers have mainly contributed with controlled clinical trials concerning both primary and secondary prophylaxis. Furthermore, they have dealt with pathophysiologic, clinical and experimental studies concerning portal haemodynamics and the evolution and treatment of variceal bleeding. CONCLUSION: The Danish studies have been well designed and are frequently cited. Further prospective randomized studies in the new treatment modalities are encouraged. PMID- 8726273 TI - Peptic ulcer pathophysiology: acid, bicarbonate, and mucosal function. AB - The previously accepted role of gastric acid hypersecretion in peptic ulcer disease has been modified by studies showing no correlation between acid output and clinical outcome of ulcer disease, or between ulcer recurrence rate after vagotomy and preoperative acid secretion. At the same time, studies have been unable to demonstrate increased acidity in the duodenal bulb in patients with duodenal ulcer, and consequently more emphasis has been given to the mucosal protecting mechanisms. The existence of an active gastric and duodenal mucosal bicarbonate secretion creates a pH gradient from the luminal acid to near neutrality at the surface of the epithelial cells, thereby acting as an important mucosal defence mechanism. The regulation of bicarbonate secretion is a complex process related to motility and neural activity. Stimulation is by acid, PGE2, NO, VIP, cAMP, and mucosal protective agents. Bicarbonate secretion is inhibited by atropine, muscarinic antagonists, alpha-adrenoceptor agonists, indomethacin, bile acids, tobacco smoking, and probably also by infection by Helicobacter pylori. Apart from mucus and bicarbonate, the mucosal defence is supported by a hydrophobic epithelial lining, rapid cell removal and repair regulated by epidermal growth factor. Sufficient mucosal blood flow, including a normal acid/base balance, is important for subepithelial protection. In today's model of ulcer pathogenesis, gastric acid and H. pylori work in concert as aggressive factors, with the open question being: why does only a fraction of the infected population develop an ulcer? PMID- 8726274 TI - Vagotomy. AB - Peptic ulcer disease was for years a common indication for surgery in Danish hospitals and considerable experience in partial gastrectomy was gained. In spite of an unquestionable mortality rate and a number of patients having postgastrectomy complaints, results were generally recognized as acceptable. Danish surgeons were for long reluctant to take up vagotomy and drainage as a primary ulcer operation, but when they did start a large number of procedures were performed. In fact, the use of this treatment culminated during two to three decades. However, on a basis of experiences from these years, Danish research contributed actively to the international evolution of the surgical vagotomy technique, the evaluation of clinical results and the studies of postoperative alterations in gastric physiology. References are selected from an extensive literature and are in no way complete. PMID- 8726275 TI - Bleeding ulcer. AB - A review is presented on theory and practice in the handling of gastroduodenal bleeding in the presurgical era with emphasis on the Meulengracht treatment. A description is thereafter given of the development of criteria for emergency surgery in the preendoscopic period with special mention of the circulatory evaluation using measurements of central venous pressure. A survey is given on the development in endoscopic diagnosis and treatment with a critical evaluation of the predictive value of clinical factors and endoscopic stigmata of haemorrhage. Finally the sparse knowledge on the haemostatic mechanisms of current endoscopic therapies is presented. PMID- 8726276 TI - Gastrointestinal endocrinology. AB - During the last quarter of this century gastrointestinal endocrinology has grown explosively. In 1970, three hormones (secretin, gastrin, and cholecystokinin) were identified and by authorities in the field considered sufficient to explain the entire hormonal regulation of digestion. That was some underestimation. Today the gut is known to express more than 20 different hormonal/regulatory peptide systems. Their widespread cellular occurrence, gene expression cascades, secretory mechanisms, receptors and receptor binding, as well as normal and pathophysiological effects are now also fairly well known owing to the marked progress in basic sciences and biochemical technologies (immuno and peptides chemistry, molecular and cell biology). Thus, the gut is now recognized as the largest endocrine organ of the body; and a substantial part of the gastroenterologic research over the latest decades has been devoted to gut hormones. The following review describes the recent development, with emphasis on gastrointestinal peptide systems that have been studied and even discovered in Denmark. Hence, as reflected by the number of doctoral theses and PhD studies (> 50 since 1974), gastrointestinal endocrinology has been a major research area in this country in the past 25 years. PMID- 8726277 TI - Gastroenteropancreatic tumours and prohormones. AB - The structures and post-translational maturation of pancreatic and gastrointestinal prohormones are reviewed with emphasis on Danish contributions to today's knowledge. The review describes general, cell-specific, and tumour specific prohormone-processing patterns. Since prohormone-processing in endocrine tumours is often attenuated, conventional assays that measure only the phenotypic endpoint of hormone gene expression (i.e. the bioactive hormone) do not quantitate tumour activity accurately. In contrast, measurements that include also prohormones and processing intermediates provide more accurate data on hormone synthesis in gastroenteropancreatic endocrine tumours. In order to comply with such demands we have developed a new analytical principle (processing independent analysis (PIA)) which quantitates the entire translation product irrespective of the degree of processing. The significance of PIA in routine diagnostics awaits prospective evaluation. We hope that the present review illustrates how the tumour biology of endocrine cells in the pancreas and the gut has been an essential research area in Danish gastroenterology and endocrinology- one purpose being improvement of early diagnosis of endocrine tumours in the gut and the pancreas. PMID- 8726278 TI - Sphincter of Oddi motility. AB - BACKGROUND: Technical improvement in endoscopic and manometric technique has allowed direct manometry of the human sphincter of Oddi (SO). The aim of the present review is to describe the present status of physiologic and clinical knowledge of the SO, with emphasis on contributions from Danish Gastroenterology. RESULTS: The SO is a zone with an elevated basal pressure with superimposed phasic contractions. It acts mainly as a resistor in the regulation of bile flow. Neurohormonal regulation influences the motility pattern. The contractions are under the control of slow waves. Clinical subgroups show abnormalcy in SO manometric pattern especially in patients with biliary or pancreatic pain without demonstrable organic substrate. Evidence suggests that endoscopic sphincterotomy may be of benefit in these patients. PMID- 8726279 TI - Chronic pancreatitis. AB - Chronic pancreatitis is a serious disease with many yet unsolved problems, e.g. pathogenesis, cause of pain and treatment. Danish gastroenterologists have for many years participated actively in the investigation of the disease and have produced many internationally recognized results, especially regarding secretion physiology and pathophysiology, epidemiology, cause of pain and characterization of the secondary diabetes mellitus. In the past 25 years more than 60 Danish papers about chronic pancreatitis have been published in international, reviewed journals. Furthermore six theses on subjects related to chronic pancreatitis have been produced. In this article the Danish contribution to the literature on chronic pancreatitis during the past 25 years is reviewed. PMID- 8726280 TI - Direct cholangiography and biliary drainage. AB - Direct cholangiography by percutaneous transhepatic cholangiography or endoscopic retrograde cholangiography has greatly improved diagnostic work-up of patients with known or suspected biliary obstruction. These diagnostic procedures were introduced in Denmark in the early 1970s, and technical refinements and clinical research of the methods were initiated. The Danish contribution led to definition of indications for direct cholangiography and general acceptance of the methods in daily clinical practice; nationally as well as internationally. The transhepatic cholangiography with selective catheterization of the biliary ducts permitted external drainage of obstructed ducts. The disadvantages of this technique inspired the innovation of internal biliary drainage and the invention of the biliary endoprosthesis. The endoscopic approach to the biliary tract and the technical improvements of accessory instruments led to the early introduction of therapeutic procedures, i.e. papillotomy, stone removal, biliary drainage and treatment of strictures and post-traumatic lesions. Experimental and clinical research with endoprostheses improved their function and prevented dislodgment. Clinical research documented that biliary drainage by endoprosthesis is a valuable alternative to surgical bypass in patients with inoperable biliary obstructions. Endoscopic therapeutic procedures for common bile duct stones have almost replaced conventional surgical treatment. Endoluminal imaging techniques are under evaluation and may contribute to future improvements. PMID- 8726281 TI - Bile acids in health and disease. AB - Over the last quarter of a century Danish research on bile acids has comprised studies of their physical and chemical properties, their physiology, pathophysiology, metabolism, and kinetics, and their clinical applicability. In the beginning of the period a major contribution was made to the understanding of the factors involved in the solubility of cholesterol in bile. The growing international understanding of the potential importance of the bile acids in health and disease gave raise to a substantial Danish contribution in the 1970s and 1980s in parallel with international achievements. Emphasis was on the possible clinical implications of bile acids. Studies on physiology and pathophysiology were in focus. Patients who have had an intestinal bypass operation for obesity served as a model for obtaining new knowledge on various aspects of the properties of the bile acids. Also the analytical methods were improved. Important physiological research on the mechanisms of hepatic bile flow was conducted. An intestinal perfusion model served as a tool providing information on absorption kinetics and on transmucosal water and electrolyte movements. The gallstone disease, liver diseases, inflammatory bowel disease, fat malabsorption, and other intestinal disorders were studied. The 'idiopathic ileopathy' as a cause for bile acid malabsorption causing diarrhoea was established as a new disorder. Thus, in the time period concerned, substantial Danish contributions emerged on major and minor topics of the bile acid field. PMID- 8726282 TI - Pacemaker cells in the gastrointestinal tract: interstitial cells of Cajal. AB - Interstitial cells of Cajal (ICC) were described a century ago as primitive neurons in the intestines. Through the years, ICC have been mistaken for neurons, glial cells, fibroblasts, smooth muscle cells, and macrophages. We identified ICC in the musculature of mouse small intestine by their characteristic morphology and topography, and we analysed the relation between ICC, autonomic nerves, and smooth muscle. Subsequent morphological and electrophysiological evidence has strongly supported our hypotheses that some ICC populations are gut pacemakers and may hold other fundamental regulatory functions (coordinative, mechanoreceptive, mediating nervous input). Recognition of common principles of ICC organization (confinement to specific locations in relation to smooth muscle layers; formation of extensive cellular networks through tight coupling of overlapping thin processes; innervation patterns; characteristic patterns of contact with smooth muscle cells) and ultrastructure (myoid features: basal lamina, caveolae, rich in sER and mitochondria, often prominent filament bundles and dense bands/bodies) has allowed the identification of ICC in the GI musculature of all species investigated. However, variation in organization and ultrastructure is significant, between both species and regions of the GI tract. Our studies of ICC in human intestine permit an extension of the above hypotheses to man and provide a basis for further studies of ICC pathology and pathophysiology. The latter may become a fruitful area of research in the coming decades. PMID- 8726283 TI - Experimental studies of intestinal ion and water transport. AB - A major advance in transport physiology was H. H. Ussing's development of the voltage-clamp method, and later the Koefoed-Johnsen-Ussing model for Na+ transport. In the same decade, J. C. Skou identified the Na(+)-K(+)-ATPase, which maintains the Na+ and K+ gradients that drive most epithelial transport processes. With this foundation, Danish scientists have pursued the mechanism of ion transport and the resulting solute-linked water flow. Recent contributions have been on isosmotic transport, suggesting solute recycling, and KCl-water cotransport in the basolateral epithelial cell membrane. Efficient small intestinal nutrient absorption is dependent on coupling to the Na+ gradient. Cotransport of Na+ and glucose is quantitatively the most important absorptive mechanism in the small intestine, as illustrated by the success of oral rehydration solutions in diarrhoea. The majority of amino acids are likewise transported by Na+ dependent carriers, but recent experiments have identified a concomitant Cl- dependency for some. Regulation of intestinal secretion, both under normal digestive processes, and in response to enterotoxins, has turned out to be very complex. It involves local and central neuronal regulation through an array of neurotransmitters and local actions of gastrointestinal hormones. Major effectors are the submucosal neurons and the main transmitters serotonin, vasoactive intestinal peptide, acetylcholine, substance P, and neurotensin. Development of antisecretagogues is impeded by the existence of several receptor subtypes and significant species differences. The Na+ and water-conserving properties of the large intestine have been shown to be regulated by adrenocortical hormones, with aldosterone as a potent stimulator of colonic Na+ absorption. A major colonic function is the symbiosis with the anaerobic bacterial population. The fermentation of carbohydrate to short-chain fatty acids, which can be absorbed, supplements small intestinal digestive function. PMID- 8726284 TI - Disaccharide digestion and maldigestion. AB - All food carbohydrates are hydrolysed to monosaccharides before transport across the microvillus membrane. The digestion of disaccharides and some oligosaccharides is undertaken by a number of small intestinal brush border enzymes: sucrase-isomaltase, lactase phlorizinhydrolase, maltase-glycoamylase and trehalase. The distribution of the enzymes in the small intestine has been investigated. Different disaccharide maldigestion syndromes have been described. Lactase deficiency in adults is a condition found in the majority of inhabitants of the world. However, the prevalence varies widely between different populations. Sucrase-isomaltase deficiency is a very rare congenital condition except in Greenland. Trehalose maldigestion is likewise rare outside Greenland. Different hypotheses regarding the molecular background of the maldigestion syndromes are discussed. PMID- 8726285 TI - Nutrition in short-bowel syndrome. AB - Short-bowel syndrome is a state of severe malabsorption secondary to extensive bowel resection. The most common reasons for extensive bowel resection are Crohn's disease and mesenteric infarction. The pathophysiological consequences depend on extent and site of resection, integrity and adaptation of the remaining bowel, and secondary effects on other organs. Most extensively bowel resected patients can be adequately nourished by mouth, especially since they develop compensatory hyperphagia. For patients with colon in function a high-carbohydrate low-fat diet is beneficial compared to a diet with a normal fat content, because it results in decreased diarrhoea, decreased faecal mineral losses, and increased energy assimilation. The relative amount of dietary fat does not influence stool mass or energy assimilation in jejunostomy patients. Patients with jejunostomy have a high faecal output of water, sodium, and divalent cations, and they often need permanent parenteral supply of saline as well as calcium and magnesium if their small intestinal remnant is < 200 cm and parenteral nutritional support if they retain < 100 cm small bowel. In contrast, 50 cm of the jejunum often suffices for adequate oral nutrition if most of the colon is preserved. The majority of patients needing long-term intravenous supply are trained to administer parenteral nutrition at home (HPN). Most patients on HPN obtain a good or fair quality of life with hospital readmissions corresponding to an average of 10% of the HPN duration and an overall HPN related mortality of about 4%. PMID- 8726286 TI - Short-chain fatty acids in the human colon: relation to gastrointestinal health and disease. AB - Fermentation, the process whereby anaerobic bacteria break down carbohydrates to short-chain (C2-C6) fatty acids (SCFAs), is an important function of the large bowel. SCFAs constitute approximately two-thirds of the colonic anion concentration (70-130 mmol/l), mainly as acetate, propionate, and butyrate. Gastroenterologists have, in spite of these facts, addressed this scientific field surprisingly late, in contrast to veterinarians, for whom the fermentative production of SCFAs has been acknowledged as a principal mechanism of intestinal digestion in plant-eating animals for decades. Interest in the effects of SCFA production on the human organism has been growing rapidly in the last 10 years, because gastrointestinal functions and beneficial effects are associated with these acids. SCFAs are of major importance in the understanding of the physiological function of dietary fibre and their possible role for colonic neoplasia. SCFA production and absorption are closely related to the nourishment of the colonic mucosa and sodium and water absorption, and mechanisms of diarrhoea. Patients with severe malabsorption compensate by the fermentation of otherwise osmotic active saccharides to SCFAs, which are readily absorbed and used as energy fuels in the organism. SCFA production from dietary carbohydrates is a mechanism whereby considerable amounts of calories can be salvaged in short bowel patients with remaining colonic function if dietary treatment is adjusted. SCFA enemas are a new and promising treatment modality for patients with ulcerative colitis. The effect has been attributed to the oxidation of SCFAs in the colonocytes. An impressive number of papers have described the effects of butyrate on various cell functions, the significance of which is still unknown. Up until now, attention has been related especially to cancer prophylaxis and treatment. Diminished production of SCFAs appears to be involved in antibiotic associated diarrhoea, diversion colitis, and possibly in pouchitis. The interaction between bacterial fermentation, ammonia metabolism, and bacterial growth and protein synthesis appears to be the main mechanism of action of lactulose treatment in hepatic coma. Pathological and extremely high rates of saccharide fermentation explain the severe deterioration in patients with D lactate acidosis. Hence, this scientific field has come late to clinical working gastroenterologists, but as work is progressing the production of SCFAs in the large bowel becomes involved in several well-known intestinal disorders. PMID- 8726287 TI - Mediators of inflammation in chronic inflammatory bowel disease. AB - A distinguishing feature of inflammatory bowel disease (IBD) is its apparently spontaneous, chronic relapsing course. Despite extensive research over several decades the etiology of IBD remains unknown, but evidence has accumulated to suggest that the mucosal inflammatory response may be caused by (i) a defective mucosal barrier function resulting in an abnormally increased exposure to luminal antigens and toxins, (ii) an appropriate immunologic response to an unusual infection, antigen or toxin, or (iii) an inappropriate immunological response to ubiquitous antigens or stimuli. In recent years, the identification of established and potential mediators of inflammation has expanded to include eicosanoids, platelet activating factor, biogenic amines, kinins, complement derived peptides, chemotactic peptides, cytokines, neuropeptides, and reactive metabolites of oxygen and nitrogen. Thus, the study of the inflammatory process has become ever more complex. Until the predisposing and trigger factors have been identified the achievement of a more rational and effective approach to therapy in IBD relies on interruption of the mechanisms responsible for excess mediator formation. As summarized in this review on the role of soluble mediators of inflammation, several Danish gastroenterologists have been profoundly engaged in basic and clinical research in the past 25 years to place some pieces of the confusing puzzle of IBD. PMID- 8726288 TI - Prophylaxis against colorectal cancer. AB - Colorectal cancer is diagnosed in more than 3000 people every year in Denmark, with a population of 5 million, and 2000 die from this disease every year. The aetiology of the disease is complex, but an increasing number of cancers have been related to genetics and Denmark is contributing with a well-established register of familial adenomatous polyposis and a recently founded register for hereditary nonpolyposis colorectal cancer, both with major international relationships. The Danish tradition of epidemiology and clinical trials has also been demonstrated in population screening trials for colorectal cancer in average risk persons as well as high-risk groups with precursors of the disease. The present review places Danish contributions within the prophylaxis of colorectal cancer during the last decade in an international context. PMID- 8726289 TI - Physiology and pathophysiology of anal function. AB - A review of the Danish contributions to the increased understanding of anorectal physiology and pathophysiology during the last 25 years is presented. In this period there has been a vast international increase in interest in anorectal physiology and pathophysiology, with much improvement in the understanding and treatment of anorectal functional disorders. The application of new sophisticated techniques to anorectal physiology research continues to improve our knowledge of anorectal function. PMID- 8726290 TI - Clinical pharmacology in gastroenterology: development of new forms of treatment of inflammatory bowel disease. AB - Kinetic-dynamic aspects of the development of slow-release mesalazine, Pentasa (now an established treatment of inflammatory bowel disease (IBD)), and cyclosporin, a T cell selective immunosuppressant (still in the investigative phase), are reviewed as examples of Danish contributions at an early stage to international, clinical drug research. Apart from increasing the therapeutic options for patients with IBD, current and future studies with these (and other) drugs may add important clues to a more precise understanding of the basic pathogenetic mechanisms (e.g. cytokines, adhesion molecules) involved in these diseases. The future development and clinical implementation of novel drug designs in IBD and other gastrointestinal diseases may be expected to benefit from a continued or even closer collaboration between clinical gastroenterologists and basic research institutions, including the pharmaceutical industry at an early stage. PMID- 8726291 TI - The randomized controlled clinical trial in gastroenterology: the Danish contributions from 1970 to 1994. AB - More than 200 Danish randomized controlled clinical trials in gastroenterology published from 1970 to 1994 were retrieved by electronic media, by hand-searching relevant journals, and by direct requests to Danish gastroenterologists. With the historical perspective through a quarter of a century, these papers are outlined to provide a survey of the pieces of information that Danish gastroenterologists have contributed to the present knowledge of therapeutics. The presented randomized controlled clinical trials constitute an impressive sum of knowledge within a diversity of topics. A cautious analysis of the time pattern for the publications in addition to the contents of the reports discloses that the discipline of planning and executing relevant controlled clinical trials is now in blossom in Danish gastroenterology. PMID- 8726292 TI - Epidemiology in gastroenterology. AB - A review of the literature on gastrointestinal epidemiology during the last 25 years shows a comprehensive contribution from Danish studies. This is partly explained by very favourable conditions in Denmark for epidemiological research, due to various valid registers and a long tradition for longitudinal population and patient cohorts. Data are presented on prevalence, incidence, demography, clinical risk factors, clinical course, and prognosis of various gastrointestinal disorders. Main emphasis is put on Danish studies, which are compared with international results. Present possibilities for prophylaxis against gastrointestinal diseases and delimitation of treatment of benign diseases are discussed. It is expected that research within gastrointestinal epidemiology in Denmark will increase in the coming years. PMID- 8726293 TI - Gastroenterology in the last decennia and initiatives in medical ethics: coincidence or linkage? AB - A quarter of a century ago ethics was an esoteric term, known to theologians and philosophers, but unknown as a discipline to the majority of doctors. Since then, however, ethics has become a substantial part of clinical medicine and health research. Ethics as an area of interest for Danish gastroenterologists appeared from several foci in the early days. One angle was an almost revival-like interest in research methodology and its ethical dimensions. Other angles were derived from Danish gastroenterologists' experiences transferred from other disciplines before the birth of Danish gastroenterology. From the time of these early incentives Danish gastroenterologists have constituted a platform for the implementation of the basic principles, lying behind medical ethics, now in collaboration with other parts of the medical profession. The topics are reflected in a number of publications and in the various practical diversions. An interest in information of patients appeared at an early stage. Publication ethics as a subdiscipline involved Danish gastroenterologists and has led to contributions within the framework of the International Group of Medical Journal Editors. Research ethics, a central topic throughout all years, has led to such important initiatives as the Second Helsinki Declaration and the establishment of a national control system for medical research in man. A further ramification of ethics is scientific dishonesty and good clinical practice. Here a recent initiative has led to the establishment of a national Committee on Scientific Dishonesty. Under the auspices of the OMGE (Organisation Mondiale de Gastroenterologie) Danish gastroenterologists have investigated transnational and transcultural differences in gastroenterologists' attitudes to information of patients and relatives and have unmasked considerable and important differences throughout the world. Medical ethics has, together with scientific methodology, to some extent reunited the sub-specialized fragments of the mother disciplines medicine and surgery and in this way has acted as partes pro toto. PMID- 8726294 TI - Prophylaxis against postoperative complications in gastroenterology. AB - Gastrointestinal surgery results in pain, profound endocrine metabolic changes and organ dysfunction, immunosuppression and decreased resistance to infection, fatigue and convalescence. The main pathogenetic mechanism is the surgical stress response, which may be reduced by minimal invasive (laparoscopic) surgical techniques and afferent neural and perhaps humoral mediator blockade. Subsequently, these techniques have been documented as reducing a variety of postoperative morbidity parameters. A unifying concept for control of the postoperative period is presented as a combined effort to enhance preoperative information, stress reduction and sufficient functional pain relief allowing early mobilization and oral nutrition. Preliminary data, in combination with laparoscopic surgery, suggest that this approach improves outcome significantly. PMID- 8726295 TI - Computer-assisted diagnosis in gastroenterology. AB - During the last 20 years, Danish gastroenterologists and biostatisticians have been involved in practical and theoretical research aimed at providing computer assisted decision support in clinical practice. On the practical side, we summarize two clinical applications, one in the acute abdomen, the other in jaundice. The former project is part of a worldwide effort which appears able to reduce considerably the perforated appendix rate and the negative laparotomy rate. In the latter project, also entwined in international activities, three methods of early statistical discrimination of jaundice causes have been tested, and it has been shown how ultrasonographic data can be incorporated into the diagnostic assessment. On the theoretical side, a Dutch-Danish group has been looking into how one best designs a trial and analyses its data in the context of measuring the quality and impact of machine-made diagnostic advice. Having pointed out the international ramifications of these activities, we outline what may lie ahead: Gastroenterologists must be prepared to join large-scale field trials of clinical advice-giving software. In the future, however, such software will often become embedded in general hospital recordkeeping systems. PMID- 8726296 TI - Carbonic anhydrase in the alimentary tract. Roles of the different isozymes and salivary factors in the maintenance of optimal conditions in the gastrointestinal canal. PMID- 8726297 TI - A new pharmacologic treatment of functional gastrointestinal disorder. A double blind placebo-controlled study with mianserin. AB - BACKGROUND: The efficacy of unselected monoamine reuptake inhibitors (tricyclic antidepressants) in the treatment of patients with functional gastrointestinal disorders (FGD) has not been convincingly demonstrated. We investigated the efficacy of an antidepressant (mianserin) with a different receptor profile (combined 5-hydroxytryptamine-2 + 3 and alpha-2 antagonist) in FGD. METHODS: After excluding patients with psychopathology and initial placebo responders from the study, eligible patients (n = 49) were randomized to 7 weeks of double-blind treatment with either mianserin, 120 mg/day, or placebo. Efficacy was assessed by using observer-completed ratings, the Global Improvement Scale, and patient self ratings, Visual Analog Scale, and Disability Scales. RESULTS: Patients taking mianserin reported less abdominal pain, symptoms of abdominal distress, and functional disability than those given placebo (p < 0.001). The efficacy was significant across different lengths of illness periods and types of functional disorder. There was no major change 4 weeks after tapering. CONCLUSION: Mianserin may be an effective and well-tolerated pharmacologic short-term treatment for functional gastrointestinal disorders in patients with no clinical evidence of psychopathology. PMID- 8726298 TI - Protective effect of teprenone on blood flow and incidence of histologic lesions in rat gastric mucosa after hemorrhage and retransfusion. AB - BACKGROUND: The effects of teprenone (6,10,14,18-tetramethyl-5,9,13, 17 nonadecatetraen-2-one) on changes in gastric mucosal blood flow, adenosine triphosphate (ATP) content, and incidence of histologic lesions were evaluated in rat gastric mucosa after hemorrhage and retransfusion. METHODS: Teprenone (100 mg/kg) was administered orally once a day for 3 consecutive days. On the 3rd day hemorrhage was induced, withdrawn blood (retransfusion) was returned, and the above variables were determined. RESULTS: Teprenone significantly inhibited the decreases in blood flow and index of mucosal oxygen saturation (ISO2) during hemorrhage in the corpus and antral mucosa. However, no effect of teprenone was observed on systemic blood pressure and ATP levels after hemorrhage and retransfusion. Teprenone significantly (p < 0.05) decreased both the incidence of ischemic lesions and the increase in the severity of lesions after retransfusion in both mucosal regions. CONCLUSION: From these results, it is concluded that the protective effect of teprenone on blood flow was partly responsible for its inhibitory effect on the incidence of lesions in the rat stomach in this hypovolemic shock model, although the former effect might be not a direct effect on systemic vascular tone. PMID- 8726299 TI - The Helicobacter felis mouse model in assessing anti-Helicobacter therapies and gastric mucosal prostaglandin E2 levels. AB - BACKGROUND: The aims of the present study were to assess the usefulness of the Helicobacter felis mouse model in the evaluation of antimicrobial therapies and the effect of Helicobacter infection on gastric mucosal prostaglandin E2 release. METHODS: Barrier-maintained BALB/c mice were infected with H. felis and treated with different antibacterial therapies. H. felis status was determined by bacterial culture, urease test, and bacterial and histologic stainings. Release of prostaglandin E2 from the gastric mucosa was measured by radioimmunoassay. RESULTS: All triple-treated mice were cleared of bacteria both 24 h and 1 month after treatment. However, tinidazole alone also resulted in 100% eradication. Monotherapies with erythromycin acistrate, tetracycline, colloidal bismuth subcitrate, and nitecapone failed to eradicate the bacteria. The release of gastric prostaglandin E2 was doubled in the infected mice (554 +/- 39, mean +/- SE) compared with the noninfected mice (270 +/- 35) (p < 0.01). CONCLUSIONS: The H. felis mouse model proved satisfactory for assessing both anti-Helicobacter therapies and the prostaglandin E2 release. The reliability of this method was improved when several methods to assess the H. felis status were used in parallel. PMID- 8726300 TI - Faecal calprotectin shedding after short-term treatment with non-steroidal anti inflammatory drugs. AB - BACKGROUND: Increased faecal calprotectin shedding indicates gastrointestinal mucosal inflammation. METHODS: We studied the effect of short-term treatment with non-steroidal anti-inflammatory drugs (NSAIDs) on faecal calprotectin shedding in two randomized crossover studies, with treatment regimens of indomethacin or naproxen for 14 days in the first study (n = 16) and lornoxicam or naproxen for 7 days in the second study (n = 18). RESULTS: The method's reproducibility and stability were satisfactory. Indomethacin and naproxen increased the faecal calprotectin significantly from a base line of 4.7 mg/l to 9.0 mg/l and 8.0 mg/l, respectively. Lornoxicam failed to increase the faecal calprotectin. Shedding after 7 days of naproxen treatment was positively correlated to gastroduodenal mucosal inflammation assessed by endoscopy. CONCLUSIONS: Although seemingly influenced by concurrent upper airway infections, the study indicates that the calprotectin test may be useful for monitoring the inflammatory response to NSAID treatment, even in short-term setting. PMID- 8726301 TI - Reduced glutamine content in colonic polyps. AB - BACKGROUND: Glutamine is the most abundant amino acid in the body. It has a key role in nitrogen metabolism and is a major source of energy for the enterocyte and many other cells. Glutamine is also essential for tumor growth, and marked changes in organ glutamine metabolism are characteristic in cancer patients. METHODS: We have investigated the catabolism of glutamine in a classic premalignant condition: the colonic adenomatous polyp. The content of glutamine and activity of two catabolic enzymes, glutamine transferase and phosphate dependent glutaminase, were studied in normal colon and in polyp mucosa. RESULTS: Free glutamine content and activity of glutaminase were significantly lower in polyps than in their adjacent mucosa. Glutamine transferase activity was significantly lower in polyp mucosa than in normal colon controls. CONCLUSIONS: Adenomatous polyps might behave as a glutamine trap, channeling glutamine to protein and nucleic acid synthesis. These changes in glutamine catabolism could play a role in colonic neoplasia pathogenesis. PMID- 8726302 TI - Growth hormone did not enhance mucosal hyperplasia after small-bowel resection. AB - BACKGROUND: Insulin-like growth factor (IGF)-I enhances the mucosal hyperplasia that normally occurs after massive small-bowel resection. The present studies examined whether growth hormone (GH) infusion increases serum IGF-I levels and, thereby, enhances mucosal hyperplasia after small-bowel resection. METHODS: Male Sprague-Dawley rats weighing approximately 140 g were subjected to 80% jejunoileal resection or sham operation (ileal transection). A mini-osmotic pump was then inserted under the skin immediately after surgery to deliver either vehicle or 3mg/kg/day GH. All animals were killed 7 days postoperatively, and the remaining intestine was removed and divided at the anastomotic site. RESULTS: Food intake did not differ between control and GH-infused groups. Resected rats infused with GH grew faster than resected rats infused with vehicle. However, GH did not stimulate the weight gain of sham-operated rats. In both duodenojejunum and ileum, GH infusion did not stimulate hyperplasia beyond that which normally occurs after small-bowel resection, nor did it increase mucosal mass in sham operated rats. GH infusion did not alter serum IGF-I or IGF-binding protein (IGFBP) levels in either resected or sham-operated rats. GH infusion resulted in increased sucrase and maltase activities in the ileal mucosa of resected rats. CONCLUSIONS: The present results suggest that a pharmacologic dose of GH directly stimulates growth of resected animals and ileal sucrase and maltase activities without increasing serum IGF-I levels or increasing absorptive surface area in the small intestine. PMID- 8726303 TI - Incidence of Crohn's disease in four counties in southeastern Norway, 1990-93. A prospective population-based study. The Inflammatory Bowel South-Eastern Norway (IBSEN) Study Group of Gastroenterologists. AB - BACKGROUND: Standardized criteria for Crohn's disease (CD) have only recently been developed, and prospective community-based incidence studies have been performed only during the past 3 decades. Geographic variations in incidence may therefore be due to differences in study design. METHODS: From 1 January 1990 to 31 December 1993 all new cases of CD in four counties in southeastern Norway were prospectively registered. RESULTS: A total of 225 new cases yielded an annual incidence of 5.8/10(5), with the highest incidence in mixed rural-urban areas. A peak of 11.2/10(5) in the annual incidence was found for the age group 15 to 24 years, with no significant differences in the overall annual incidence by gender. An average duration of 6 months of disease before diagnosis was unchanged during the 4 years. About half of the patients had isolated colonic disease, and one quarter had isolated small-bowel disease. CONCLUSIONS: This study confirms the high incidence figures for Scandinavia, with a particularly high incidence in mixed rural-urban areas. Ileocolonoscopy improves the accuracy of the diagnosis and of the determination of disease extent, which may have therapeutic implications for the treatment and follow-up of patients. PMID- 8726304 TI - Incidence of ulcerative colitis and indeterminate colitis in four counties of southeastern Norway, 1990-93. A prospective population-based study. The Inflammatory Bowel South-Eastern Norway (IBSEN) Study Group of Gastroenterologists. AB - BACKGROUND: The incidence of ulcerative colitis (UC) has been difficult to interpret because prospective studies have only been performed during the past 3 decades. Geographic variations may therefore be due to differences in study design. METHOD: From 1 January 1990 to 31 December 1993 all new cases of UC in four counties in southeastern Norway were prospectively registered. Cases diagnosed as indeterminate colitis (IND) when endoscopy and histopathology were inconclusive or diverged with regard to diagnosis of UC or Crohn's disease (CD) were also included in the study. RESULTS: A total of 525 cases of UC and 93 cases of IND yielded an mean annual incidence of 13.6/10(5) and 2.4/10(5), respectively. There were differences in incidence between counties, and a peak of 21.5/10(5) in the annual incidence was found for the age group 25 to 34 years in UC. The distribution was about equal for each of the groups proctitis and left sided and extensive colitis. The time interval from onset of symptoms to diagnosis was 4 months. CONCLUSION: In this study one of the highest incidences of UC in the world has been found. The classification 'indeterminate colitis' seems reasonable to use in some of the cases to prevent misclassification at the initial stage of diagnosis. PMID- 8726306 TI - Mortality in patients with Crohn's disease. AB - BACKGROUND: No data on mortality for Crohn's disease are available from southern Europe. METHODS: Five hundred and thirty-one patients with Crohn's disease were observed in our unit between 1973 and 1993. In 325 patients the first diagnosis was made in our hospital. In this consecutive incidence series, in which the follow-up was 99% complete, the standardized mortality rate (SMR) was calculated. RESULTS: Nine deaths were observed, against 9.25 expected. The SMR was 0.97 (95% confidence interval (CI), 0.4-1.8). The relative risk of dying was significantly higher in the female group in the first 5 years after diagnosis (SMR, 10.3; 95% CI, 2.30-30.2). There was an excess of deaths from tumors of the digestive organs (1 observed, 0.37 expected). CONCLUSIONS: These results show that in our geographic area the mortality from Crohn's disease was not increased as shown in other community studies. PMID- 8726307 TI - Adenoma-carcinoma sequence in the papilla of Vater. AB - BACKGROUND: Our aims were to investigate the question as to whether the adenoma carcinoma sequence also applies to the papilla of Vater and to determine the frequency with which a tumour of the papilla of Vater, diagnosed histologically as containing portions of an adenoma, already contains adenocarcinoma elsewhere. METHODS: A retrospective analysis was carried out of the forceps, snare biopsy, and excised specimens obtained from the papilla of Vater in 559 patients, including 210 patients with the primary diagnosis of carcinoma and 110 patients with the diagnosis of 'portions of an adenoma'. Comparisons were made of the age distribution of patients with adenoma and those with carcinoma, and the frequency of residual adenoma in a carcinoma as a function of degree of malignancy, T stage, and UICC stage. A follow-up examination of the patients with the primary diagnosis 'parts of an adenoma of the papilla of Vater' was performed. RESULTS: The average age of the patients with the final diagnosis adenoma was 65.2 years, and that of the carcinoma patients 73.0 years. Residual adenoma was found in 54.9% of the 51 surgical specimens and in 31.7% of the 159 biopsy specimens obtained from carcinomas. The frequency of residual adenoma in carcinomas decreased statistically significantly with increasing T stage and degree of malignancy. The follow-up examination of the 110 patients with the primary biopsy diagnosis parts of an adenoma of the papilla of Vater showed unequivocal results in 80 cases. Only in 39.7% of these cases was the primary diagnosis adenoma confirmed, whereas in 60.25%, further histologic examination showed a carcinoma. CONCLUSIONS: An analysis of our results, together with those reported in the literature, indicates that the adenoma-carcinoma sequence also applies to the papilla of Vater and that, in a high percentage of patients with the histologic diagnosis 'parts of an adenoma, other regions of the tumour already contain a carcinoma. PMID- 8726305 TI - Smoking is a risk factor for osteoporosis in women with inflammatory bowel disease. AB - BACKGROUND: Some patients with inflammatory bowel disease have reduced bone mineral density, but the risk factors for osteoporosis in these patients are unclear. METHODS: To evaluate the effect of smoking and other lifestyle factors on bone mineral density in patients with inflammatory bowel disease, we studied 67 patients with ulcerative colitis, 78 with Crohn's disease, 7 with indeterminate colitis, and 73 healthy control subjects. Bone mineral density of the lumbar spine and the proximal femur was measured, using dual-energy X-ray absorptiometry. Measures of smoking and other lifestyle factors were assessed in an interview. RESULTS: The female ex- or current smokers with inflammatory bowel disease (n = 38) had lower age- and sex-adjusted Z-scores of bone mineral density than the female patients who had never smoked (n = 34) (Z-scores in the lumbar spine, -0.277 (1.283) (mean (standard deviation)) and 0.487 (1.056), respectively; p = 0.008; and in the femoral neck, -0.626 (1.055) and -0.013 (1.019); p = 0.015). These differences were not explained by the type or treatment of the disease, the menstrual history, or the use of estrogen preparations. In male patients no differences in bone mineral density were found between ex- or current smokers and non-smokers. Coffee drinking and alcohol consumption were not associated with bone mineral density in these patients. CONCLUSIONS: Smoking is associated with low bone mineral density in women with inflammatory bowel disease. This association is not related to the body mass index, the medical treatment, or the type of disease. PMID- 8726308 TI - Cholecystokinin-A and cholecystokinin-B/gastrin receptor mRNA expression in the gastrointestinal tract and pancreas of the rat and man. A polymerase chain reaction study. AB - BACKGROUND: Gastrin and cholecystokinin (CCK) are thought to exert trophic effects on the gastrointestinal tract and pancreas. Two types of receptors have been cloned, CCK-A and CCK-B/ gastrin. We have examined the occurrence of CCK-A and CCK-B receptor mRNA in the brain, digestive tract, pancreas, and kidney of the rat and man by Northern blot and reverse transcribed polymerase chain reaction (RT-PCR). METHODS: Total RNA was isolated from rat tissues and reverse transcribed into cDNA. cDNA from brain, kidney, and pancreas of the rat and man and from human whole stomach were commercially available. Northern blot and a PCR technique based on Taq polymerase-antibody interaction and using CCK-A and CCK-B receptor-specific primers, followed by Southern blot analysis, were the methods used. RESULTS: By means of Northern blots, CCK-A receptor mRNA was detected in rat fundus mucosa and pancreas but not in the remaining GI tract or brain. By means of RT-PCR, CCK-A receptor mRNA was demonstrated in the brain and the mucosa of the fundus, antrum, duodenum, and colon, kidney, pancreas and pancreatic islets. CCK-B receptor mRNA was detected by Northern blot analysis in the brain and the fundus mucosa but not in the rest of the digestive tract and not in the pancreas, pancreatic islets, or kidney. By RT-PCR, expression of CCK-B receptor mRNA could also be detected in antrum mucosa. In man, CCK-A receptor mRNA was detected in the brain, stomach, pancreas, and kidney, whereas CCK-B receptor mRNA was found in the brain, stomach, and pancreas but not in the kidney. Cloning and DNA-sequence analysis of the PCR-amplified rat and human CCK-A and CCK-B receptor DNA fragments, which cover the protein-encoding regions of the intracellular loop C3, showed complete sequence homology as compared with published rat and human sequences. CONCLUSIONS: It appears unlikely that CCK will have effects in the ileum, at least not effects mediated by CCK-A receptors. It also appears unlikely that physiologic concentrations of gastrin in the circulation will promote growth (or exert other effects) in the pancreas, duodenum, ileum, and colon, since CCK-B receptor mRNA is not expressed or is poorly expressed in these tissues. PMID- 8726309 TI - Effects of wortmannin, a novel myosin light-chain kinase inhibitor, on bile canalicular contraction in vitro and in vivo. AB - BACKGROUND: The cytoskeletal system is believed to play an important role in normal bile formation. The effects of wortmannin, a new myosin light-chain kinase inhibitor, on bile canalicular contraction and bile flow have been observed. METHODS: The bile canalicular contraction of cultured hepatocyte doublets was investigated, using an image analyzer with a phase contrast microscope, and the intracellular Ca2+ concentration was measured, using microscopic fluorometry. We also investigated bile flow by in vivo intraportal infusion of the drug in rats. RESULTS: Treatment with wortmannin inhibited norepinephrine-induced canalicular contraction and caused a decrease in bile flow without changing systematic and portal blood pressure. Morphologic examination of the electron microscopic study showed that most bile canaliculi were dilated, with loss of microvilli, but no other apparent damage was seen in parenchymal hepatocytes. CONCLUSIONS: These data suggest that the integrity of the phosphorylation system of myosin is essential for normal bile flow. PMID- 8726310 TI - Bacterial adherence and biofilm formation on latex and silicone T-tubes in relation to bacterial contamination of bile. AB - BACKGROUND: T-tube-related bacteriobilia causes infectious complications and obstruction. To prevent these, the choice of T-tube material may be of importance. METHODS: Transected common bile ducts (CBDs) of 17 piglets were sutured over latex or silicone T-tubes, or without a T-tube. RESULTS: After 6 weeks bacteriobilia was found in all of 12 CBDs with and in 1 of 5 CBDs without a T-tube (p < 0.05). By scanning electron microscopy (SEM) four of five latex and none of five silicone T-tubes had bacterial biofilms (p < 0.05). All tubes remained patent. Segments of T-tubes were incubated with five different bacterial strains. Sonication and SEM showed that 0.1-1.1% of 10(7) colony-forming units of inoculum adhered to T-tubes. Two to six times more bacteria adhered to latex than to silicone (p < 0.05). CONCLUSIONS: Silicone offers better long-term patency than latex. Less infectious complications occur if T-tubes are omitted. PMID- 8726311 TI - Gastrin does not stimulate growth of the rat pancreas. AB - BACKGROUND: Gastrin is thought to stimulate growth of the pancreas via gastrin/cholecystokinin (CCK)-B-type receptors. The aim of the present study was to examine the trophic response of the pancreas to exogenous gastrin or to hypergastrinemia of endogenous origin and to hypogastrinemia with or without concomitant hyperCCKemia. METHODS: Hypergastrinemia was induced in male Sprague Dawley rats by continuous infusion of human Leu15-gastrin-17 (5 nmol/kg/h, subcutaneously), by removal of the acid-producing part of the stomach (fundectomy), or by treatment with omeprazole (400 mumol/ kg/day, orally). Hypogastrinemia was induced by antrectomy or by gastrectomy. HyperCCKemia was induced by pancreaticobiliary diversion (PBD). The rats were killed 10 days or 8 weeks after the operations or treatments. The concentrations of circulating gastrin and CCK were measured by radioimmunoassay. The pancreatic weight and DNA content were determined. RESULTS: Gastrin infusion, omeprazole treatment, and fundectomy greatly increased the serum gastrin concentration. The resulting levels were very similar in the three groups and probably represent the maximum attainable physiologic serum gastrin concentration. Whereas gastrin infusion or omeprazole treatment (hypergastrinemia) and antrectomy (hypogastrinemia) were without effect on the weight and DNA content of the pancreas, gastrectomy (hypogastrinemia) and fundectomy (hypergastrinemia) increased the weight and DNA content. PBD (hyperCCKemia) greatly increased the weight and DNA content of the pancreas. PBD plus fundectomy, PBD plus gastrectomy, PBD plus antrectomy, and PBD plus omeprazole increased the weight and DNA content of the pancreas, as did PBD alone. CONCLUSION: CCK is a physiologically important trophic stimulus for the rat pancreas, but gastrin is not. The increase in pancreatic weight and DNA content after fundectomy and gastrectomy cannot be explained by means of either gastrin or CCK. PMID- 8726312 TI - Acute pancreatitis in Buskerud County, Norway. Incidence and etiology. AB - BACKGROUND: Data on the incidence of acute pancreatitis in Norway are sparse. Danish studies have shown incidence rates increasing to about 35/100,000 (26-65% alcohol-induced), higher than found in American and British studies. METHODS: We have prospectively registered all cases of acute pancreatitis during 1 year in the county of Buskerud, with a population of 224,000. RESULTS: The incidence of acute pancreatitis was 41.5 per 100,000. The median age of the patients was 63 years (range, 21-96 years). The main etiologic factors were gallstone disease (51%) and alcohol abuse (15%), whereas 10% of the cases were classified as idiopathic. Pseudocysts or abscesses developed in 8.8%. The mortality was 6.5%. CONCLUSION: The incidence of acute pancreatitis in this population is comparable to what has been found elsewhere in Scandinavia, but the relative frequency of alcoholic pancreatitis is substantially less. PMID- 8726313 TI - Esophageal malignancy after liver transplantation in a patient with Barrett's esophagus. AB - Patients undergoing liver transplantation are predisposed to develop extrahepatic malignancies. It is also known that patients with predisposed conditions, such as Barrett's esophagus, have higher rates of esophageal carcinoma. We present here a patient who underwent liver transplantation, had Barrett's esophagus, and developed esophageal malignancy a short time after transplantation. Liver transplantation may be associated with acceleration of the precancerous condition and the development of malignancies. PMID- 8726314 TI - Sports-specific injuries. AB - Injuries to the upper extremities can happen in any sport. Injury patterns are common to specific sports. Understanding which injuries occur with these sports allows the examiner to diagnose and treat the athlete easily. This article reviews some of the injuries common in sports such as bicycling, golf, gymnastics, martial arts, racquet sports, and weightlifting. PMID- 8726315 TI - The role of arthroscopy in athletic injuries of the wrist. AB - Arthroscopy has advanced our understanding of wrist dysfunction due to injuries sustained during participation in sports. Although the initial role of arthroscopy was primarily to facilitate diagnosis of specific injuries, technologic advances have allowed many of these entities to be managed arthroscopically. This minimally invasive surgery, which is used to treat cartilage lesions, ligament instability, synovitis, and other injuries, allows early and more specific diagnosis, satisfactory management, and early return to play or work. Arthroscopy should be considered a primary means of evaluation and management of athletic injuries of the wrist. PMID- 8726316 TI - Soft-tissue injuries of the wrist. AB - The wide spectrum of athletic activities places demands of different magnitudes, orientations, and degrees of repetition on the wrists of athletes. These demands can result in injuries to the soft tissues of the wrist, which may make optimal athletic performance difficult if not impossible. With the advent of increased awareness of injuries particular to a sport and advances in diagnostic acumen, both technologic and clinical, these once enigmatic pathologic entities can be approached with a treatment plan that often can return the athlete to competition quickly. A number of these injuries and their pathomechanics, diagnosis, and treatment options have been described in this article. Although adequate treatment of the subject of athletic soft-tissue injuries to the wrist requires a more lengthy discussion than is appropriate here, the material presented on dorsal pain disorders, carpal instability, and the triangular fibrocartilage complex should serve as a starting point for increasing cognizance and understanding of the injured wrist in the athlete. PMID- 8726317 TI - Arthroscopy of the elbow. AB - As our understanding of the anatomy and function of the elbow joint continues to grow and technology continues to advance, our ability to correct disorders of the elbow with arthroscopic techniques will expand. Today, we are at the brink of major advances in the arthroscopic evaluation and treatment of elbow ailments. Many open surgical procedures currently being performed will undoubtedly be adapted for an arthroscopic approach, as we are already seeing in the treatment of radiocapitellar arthrosis, tennis elbow, arthrofibrosis, and ulnohumeral arthroplasty. Elbow procedures, such as ligamentous tightening, fracture treatment with bioabsorbable devices, and biologic joint replacement will be commonly performed in the future with the aid of the arthroscope. Although elbow arthroscopy is technically demanding, it is a highly effective surgical technique in treating many intra-articular disorders with minimal morbidity. Most of the complications associated with elbow arthroscopy can be avoided by adhering to strict and proper surgical technique. Successful elbow arthroscopy requires a thorough understanding of local gross and arthroscopic anatomy. To maintain proper orientation at all times, the skin should be properly marked before starting the procedure. The joint should be kept distended during initiation of portals to move the neurovascular structures away from the arthroscopic instruments. Nonvented cannulas with blunt trocars should be used to allow for safe passage of instruments and to avoid multiple capsular punctures. Finally, the elbow should remain flexed to 90 deg during most of the procedure, thus keeping the neurovascular structures in the antecubital fossa relaxed. If these techniques are followed, the surgical morbidity should remain low, and surgeon and patient will find elbow arthroscopy tremendously effective. PMID- 8726318 TI - Medial and lateral epicondylitis in the athlete. AB - An appropriate diagnosis must be made after carefully excluding all other options in treating the patient with lateral epicondylitis. The majority of these patients will do well with nonsurgical treatment; however, if unresponsive to this regimen, a carefully selected patient will have a successful result with surgical reconstruction (85%-95% of patients). Attention to detail both pre- and postoperatively will help to create a successful result in a disease entity that plagues a large proportion of our athletic population. PMID- 8726319 TI - Elbow injuries to the throwing athlete. AB - Elbow injuries in the throwing athlete are common. Because of the tremendous medial tensile forces and lateral compressive forces borne by the elbow, there is a wide array of injuries that occur. A detailed history and physical examination are necessary for an accurate diagnosis. Many of the injuries discussed can be treated successfully with conservative measures followed by aggressive physical therapy. Prompt recognition and early treatment is mandatory. In those conditions in which conservative treatment tends to fail, the detailed surgical options have been discussed. It is important to select highly motivated patients for those procedures that require extensive rehabilitation postoperatively. The keys to success in elbow surgery are a compliant patient and a well-regimented rehabilitation program. PMID- 8726320 TI - Compressive neuropathies and tendinopathies in the athletic elbow and wrist. AB - Overuse syndromes of the upper extremity in the athletic population are a common and often difficult problem for physician and patient alike. Optimal function of the upper extremity is tied intimately to success in many sporting activities. Correct diagnosis and proper care require a thorough knowledge of the pertinent anatomy, pathophysiology, and pathomechanics involved in each disorder. Conservative care with rest, activity modification, and medication is adequate for most athletic injuries. Surgical intervention may be indicated for continuing pain, decreased performance, or to prevent chronic changes. Surgery must be followed by thoughtfully prepared training and rehabilitation programs to optimize the chances of a successful outcome. PMID- 8726321 TI - Bony injuries of the wrist, forearm, and elbow. AB - One of the greatest challenges when examining an injured athlete is ensuring that the examination encompasses the whole patient and not just the obvious deformity. Bony injuries of the wrist, forearm, and elbow are uncommon in athletes and when present should always lead to suspicion of possible concomitant soft tissue injury. Mechanisms causing osseous disruption are of fairly high energy and can be quite disabling to the long-term career of the athlete. Appropriate expectations with regard to both treatment and the possible return to sports are critical in providing successful treatment for these often complex injuries. There is no substitute for a careful clinical history and physical examination in not only establishing the diagnosis but ensuring that a complete diagnosis with its subsequent prognostic outcome can be rendered. PMID- 8726322 TI - The immature athlete. Common injuries and overuse syndromes of the elbow and wrist. AB - Specific elbow and wrist injuries are predictable in the skeletally immature athlete based on the biomechanics of the sport and the age of the patient. The physician must be aware of the potential for overuse injuries. Modification in training regimens is essential for recovery. A greater emphasis must be placed on the prevention of these injuries. As a general rule, the young athlete should not progress more than 10% per week in the amount and frequency of training. Correction of muscle-tendon imbalances is accomplished by maintaining strength and flexibility of susceptible tissues. In throwers, a triceps-strengthening program of progressive resisted extension exercises and a forearm flexor/extensor strengthening program using the French curl technique are helpful. Careful attention to throwing technique and proper coaching are essential. The goal for the young athlete is early recognition of the injury and thereby prevention of a long-term disability. PMID- 8726323 TI - Unusual problems of the athlete's elbow, forearm, and wrist. AB - Early in their medical training physicians often are told, "When you hear hoofbeats, don't think of zebras." Such sage advice is helpful when facing the formidable challenges of information acquisition early on. Later, however, we must be familiar and consider these uncommon conditions when entrusted with the responsibility of properly caring for our patient athletes. This is particularly true in the athlete presenting with symptoms or findings disproportionate to his or her injury, or when symptoms persist despite seemingly appropriate treatment. A high index of suspicion and attention to radiographs and further appropriate work-ups result in detection of most occult unexpected lesions. PMID- 8726324 TI - Cytochalasin W, a new 24-oxa[14]cytochalasan from Phoma exigua var. heteromorpha. AB - A further cytochalasin was isolated from liquid culture filtrates of Phoma exigua var. heteromorpha together with cytochalasins U and V and other well-known cytochalasins. The metabolite, named cytochalasin W, was characterized by spectroscopic and chemical methods as a new 24-oxa[14]cytochalasan, bearing a formyl group on the macrocyclic ring. Cytochalasin W showed toxic activity in the brine shrimp assay. PMID- 8726325 TI - Brazilian Baccharis toxins: livestock poisoning and the isolation of macrocyclic trichothecene glucosides. AB - Samples of the toxic Brazilian plant, Baccharis coridifolia, which is responsible for numerous cases of livestock poisoning in southern Brazil and Argentina, were collected during the growing season, and the toxicities in calves of the plant materials were correlated with the levels of macrocyclic trichothecenes present. Female plants in flower were considerably more toxic than male plants or plants not in flower. Plants not in flower were of intermediate toxicity. The female plants in flower typically contained 5-10 times the levels of toxins as were found in the male plants. In addition, six new glucosides of the macrocyclic trichothecenes were isolated and characterized. The most prominent glucosides, those of roridins A and E, were found in high levels in the female plants. PMID- 8726326 TI - Simultaneous occurrence of diarrhetic and paralytic shellfish poisoning toxins in Spanish mussels in 1993. AB - Mussel aquaculture is an important industry for the Galician Rias, located in northwestern Atlantic coast of Spain. Since 1976 this region has been seriously affected by incidents of paralytic and diarrhetic shellfish poisoning (PSP and DSP). A particularly bad episode occurred in 1993, when the toxic event lasted for an unusually long period. Many people were stricken ill with unusual symptoms. In this paper we report on the chemical analysis of toxic 1993 mussel samples, using the techniques of liquid chromatography and capillary electrophoresis coupled with mass spectrometry. These analyses revealed a very complex toxin profile, with both PSP and DSP toxins present. Two DSP toxins, okadaic acid and DTX2, were observed, while the primary PSP toxins were B1 and the decarbamoylated derivatives of saxitoxin, GTX2 and GTX3. Small amounts of saxitoxin and other as yet unidentified PSP toxins were observed. PMID- 8726327 TI - Effect of an unknown toxin isolated from Dinophysis sp.-contaminated French mussels, on the electrical and mechanical activity of frog heart. AB - The effects of an unknown toxin, isolated along with okadaic acid from the hepatopancreas of French mussels contaminated by Dinophysis sp., producing ataxia, neurologic symptoms, bradycardia, arrhythmias, electrocardiographic changes, and cardiac arrest, have been studied in terms of the electrical and mechanical activity of frog atrial heart muscle. The toxin, in a dose-dependent manner, decreased the amplitude of the stimulated peak tension of isolated fibers. The toxin (1-36 micrograms/ml) did not modify the membrane resting potential but decreased the amplitude of the plateau and shortened the duration of the action potential. The toxin inhibited the Cd-sensitive L-type Ca current and increased a 4-aminopyridine-sensitive outward current in voltage-clamped cardiac myocytes. The data show that the cardiac effect of the toxin is markedly different from that of okadaic acid. PMID- 8726328 TI - Enzymatic oxidation products of allelochemicals as a basis for resistance against insects: effects on the corn leafhopper Dalbulus maidis. AB - Oxidation products of com allelochemicals generated by peroxidases or tyrosinases were tested in 10% sucrose solutions for effects on the corn leafhopper Dalbulus maidis. Some reduction in feeding was noted with hydrogen peroxide (a cofactor for peroxidase). Significant reduction in feeding was noted with chlorogenic acid, ferulic acid, p-coumaric acid, and 6-methoxybenzoxazolinone (MBOA), but not rutin at 400 ppm in solution. Oxidation products of these compounds all caused significantly less feeding by the leafhoppers compared to the original compound. Oxidation products generated by peroxidase from ferulic acid and 6 methoxybenzoxazolinone caused significant mortality to the leafhoppers within 5 days. Thus, provided conditions are such that oxidizing enzymes and allelochemicals can interact due to damage by insects, resistance may be significantly enhanced by the oxidized products as opposed to the effects of the allelochemicals alone. PMID- 8726329 TI - Toosendanin facilitates [3H]noradrenaline release from rat hippocampal slices. AB - Slices of hippocampus of the rat, preincubated with [3H]noradrenaline ([3H]NA), were used to investigate the effects of toosendanin on the release of [3H]NA. Toosendanin potently enhanced spontaneous 3H outflow. Seventy-four percent of the enhancement was inhibited by reserpine pretreatment. The toosendanin-induced 3H overflow was in a concentration-dependent manner (5-60 microM) both in the presence and absence of extracellular calcium. Under Ca(2+)-free conditions, the effect of toosendanin on 3H outflow was unchanged by TTX, but inhibited by Ca(2+) chelator BAPTA-AM; dantrolene sodium failed to affect the toosendanin-induced 3H outflow, while 3,4-diaminopyridine showed an additive effect on the outflow with this substance. The findings suggest that in the absence of extracellular Ca2+, toosendanin enhances [3H]NA release through the liberation of intracellular Ca2+ stores. PMID- 8726330 TI - Lichen acids as uncouplers of oxidative phosphorylation of mouse-liver mitochondria. AB - Three lichen acids-namely, (+)usnic acid, vulpinic acid, and atranorin-were isolated from three lichen species (Usnea articulata, Letharia vulpina, and Parmelia tinctorum, respectively). The effects of these lichen products on mice liver mitochondrial oxidative functions in various respiratory states and on oxidative phosphorylation were studied polarographically in vitro. The lichen acids exhibited characteristics of the 2,4-dinitrophenol (DNP), a classical uncoupler of oxidative phosphorylation. Thus, they released respiratory control and oligomycin inhibited respiration, hindered ATP synthesis, and enhanced Mg(+2) ATPase activity. (+)Usnic acid at a concentration of 0.75 microM inhibited ADP/O ratio by 50%, caused maximal stimulation of both state-4 respiration (100%) and ATPase activity (300%). Atranorin was the only lichen acid with no significant effect on ATPase. The uncoupling effect was dose-dependent in all cases. The minimal concentrations required to cause complete uncoupling of oxidative phosphorylation were as follows: (+)usnic acid (1 microM), vulpinic acid, atranorin (5 microM) and DNP (50 microM). It was postulated that the three lichen acids induce uncoupling by acting on the inner mitochondrial membrane through their lipophilic properties and protonophoric activities. PMID- 8726331 TI - Free vascularized deep peroneal nerve grafts. AB - An ideal donor site for vascularized nerve grafts should have a constant anatomy, minimal functional loss after the nerve has been sacrificed, and a dependable blood supply parallel to the nerve over a relatively long distance. Creating a pedicle for a free vascularized deep peroneal nerve graft with the anterior tibial vessels seems to be a most suitable method for repairing long nerve gaps of over 20 cm and digital nerve defects with severe finger damage. Applications of this nerve graft to digital nerve losses with severely scarred beds created by avulsion injury, and two-stage reconstruction in some partial brachial plexus palsies (free vascularized nerve graft in the first stage and free vascularized muscle graft in the second stage) are well indicated. Advantages of this technique are: (1) A long nerve graft (up to 25 cm) can be obtained, and anomalies are rare (the nerve is absent in only 4 percent of cases). (2) The caliber of the vascular pedicle is large (approximately equal to 3 mm). (3) The nerve has a sufficient blood supply from the collateral blood vessels. (4) The graft can be easily obtained in the supine position. (5) A monitoring skin flap, based on the inferior lateral peroneal artery, can be attached to the nerve graft. (6) Sensory loss resulting from the sacrifice of the nerve covers a minimal area. (7) A donor scar on the anterior aspect of the lower leg is more acceptable than one on the posterior aspect because of less movement in walking. Disadvantages of this technique are: (1) Sacrifice of the large vessels in the lower leg may result in circulatory complications in the donor foot; to avoid this problem, preoperative angiography is recommended. (2) The donor scar is in an exposed area in female patients. (3) There may be temporary postoperative edema and disability in the donor leg. PMID- 8726332 TI - Functional evaluation at the medial gastrocnemius donor site in rats. AB - The transfer of a skeletal muscle from a donor to a recipient site creates an initial deficit in the structure and function of the muscle group from which it originates. Removal of the donor muscle induces hypertrophy of the remaining synergistic muscles, which compensate for part of the deficit at the donor site. The medial gastrocnemius (MGN) muscle is a frequently utilized donor muscle. Compared with the mass and force production of the control four-muscle plantar flexor group in rats, removal of the MGN muscle creates an initial deficit of approximately 36 percent. At 60, 90, and 120 days after removal of the MGN muscle, the degree of compensation of the remaining three-muscle plantar flexor group (lateral gastrocnemius, soleus, and plantaris muscles) was evaluated. The mass of the three-muscle group increased 13 percent over the time course studied, but was still 28 percent less than the mass of the control four-muscle group. Similarly, the maximum force of the three-muscle group increased 27 percent, but was 21 percent lower than the control four-muscle group. The authors propose a model that illustrates the function restored at a donor site in terms of the percentage of the total muscle group comprised by the donor muscle and the ability of the remaining muscle group to compensate for its removal. PMID- 8726333 TI - The use of cultured Schwann cells in nerve repair in a rabbit hind-limb model. AB - A 3-cm peripheral nerve gap in a rabbit hind-limb model was repaired by using a polyglycolic acid (PGA) conduit filled with a gelatin/Schwann-cell suspension. Postoperative nerve function after 16 weeks, as measured by isometric twitch and tetanic muscle strengths, and nerve conduction velocities, failed to demonstrate a statistically significant difference, compared to a control group in which the nerve gap was reconstructed by using a PGA conduit filled with gelatin only. The 3-cm gap in the described model may not have been long enough to show a significant difference between the two groups. Alternatively, the transferred cultured Schwann cells may have been functionally inactive. PMID- 8726334 TI - Functional assessment of peripheral nerve injury and repair. AB - Based on 191 follow-ups of patients with peripheral nerve injury and repair, the authors suggest that a reasonable assessment should consist of motor, sensory, and autonomic nerve indices. It is also suggested that a method of functional assessment about the autonomic nerve should be included. The method of evaluating motor function should be selected according to the site of the nerve injury. Lovett's method was adopted for nerve injury at the wrist and lower third of the forearm, as well as the British Medical Research Council (BMRC) method for nerve injury at the upper forearm or other nerves. Standards and grades of the BMRC should be raised somewhat, in order to compare more consistently with the results obtained using the method of Lovett; then both can be correlated satisfactorily. PMID- 8726335 TI - Functional evaluation of gap vs. abutment repair of peripheral nerves in the rat. AB - Various expressions of nerve regeneration specificity (tissue, target, and end organ) have been demonstrated histologically, but the influence of nerve regeneration specificity on functional outcome remains to be investigated. Specificity expression appears to be dependent, to some extent, on the distance between the proximal and distal nerve stumps. Regeneration specificity is inhibited by end-to-end alignment. This study examined the effect of gap distance repair on the return of rat hind-limb function, as measured by the sciatic function index (SFI). Sixty-four Sprague-Dawley rats were randomized into three groups: 0-mm gap (abutment), 2-mm gap, and 5-mm gap repair. The sciatic nerve was divided and secured in a Silastic conduit to maintain the gap length. The SFI was measured preoperatively and then at postoperative weeks 2, 4, 8, 12, 16, and 20. Results showed no significant difference in functional return between the abutment and the two gap groups at any time period (SFI at 20 weeks = -73.9, 0 mm; -73.5 2 mm; -75.2, 5mm; p = 0.90). This lack of difference in functional outcomes suggested an insignificant functional influence of regeneration specificity within single chamber conduits. The study also demonstrated that small gaps between nerve ends within a conduit yielded functional results equal to end-to-end repair of rat sciatic nerves. PMID- 8726336 TI - Exploring the use of the medicinal leech: a clinical risk-benefit analysis. AB - The medicinal leech, Hirudo medicinalis, has been used with increasing frequency for salvage of compromised pedicled flaps and microvascular free-tissue transfers. The data pertaining to 18 cases in which flap salvage with leeches was attempted were reviewed and contrasted with the data from 108 published cases from other centers. A further analysis isolated and examined 19 cases in which infection, secondary to leech utilization, supervened. Results suggested that successful salvage of tissue with leeches occurs in 70 to 80 percent of cases. In contrast, the infection rate of most series is between 7 and 20 percent and, when a clinically significant infection occurs, the flap salvage rate drops to 30 percent or less. In addition to infection, the risks of leech therapy include blood loss, which may require replacement transfusion, loss of leeches into body orifices and spaces, allergic reactions, and adverse psychological responses. Clinical indications and contraindications, and strategies to reduce the risks of leech usage, are discussed. Conclusions are that leeches are best used early, and in accordance with an accepted institutional protocol. For each case, the potential risks and benefits must be rigorously assessed and honestly applied. PMID- 8726337 TI - Ankle stance angle: a functional index for the evaluation of sciatic nerve recovery after complete transection. AB - This study attempted to develop a motor functional index, ankle stance angle (ASA), to assess rat sciatic nerve regeneration subsequent to autografting. ASA, 50 degrees in normal rats, is the ankle joint angle at the mid-stance phase of the gait cycle. In a nerve graft group, a 1-cm segment of the right sciatic nerve was transected and then repaired with nerve autograft. In an ungrafted group, the nerve gap was left unrepaired. ASA measured 4 months after surgery was statistically significantly larger in the nerve graft group (36 degrees) than in the ungrafted group (22 degrees). The results suggest that ASA is more sensitive than sciatic function index in detecting functional recovery after a complete sciatic nerve lesion. ASA also showed a significant correlation with the passive range of ankle joint motion and gastrocnemius muscle weight. The study concluded that ASA is a reliable index for assessment of regeneration of rat sciatic nerve after a complete lesion. The intra-rater reliability (r = 0.97 and 0.90) and inter-rater reliability (r = 0.85) tests performed support the conclusions. PMID- 8726338 TI - Venous flaps and perivenous areolar tissue: an experimental study in rats. AB - Since plastic surgeons are continually looking for new possibilities for the clinical transfer of tissue, they are very interested in investigating capillary blood flow. Recently, several authors have investigated the role of veins in experimental island venous flaps. In the present study, the authors attempt to clarify the importance of plasmatic imbibition and perivenous tissue as single nourishment sources in experimental island skin flaps deprived of arteries, and consisting of both axial and random zones. This study demonstrates the importance of perivenous areolar tissue in flap survival; the "initial plasmatic imbibition" was not able to keep the flaps viable. Due to the previously demonstrated presence of small arteries in the pedicle of "venous flaps," these flaps should no longer be considered as purely venous. In the reported rat model, it was not possible to harvest a flap without arterial inflow. PMID- 8726339 TI - Limiting impairment of muscle function following ischemia and reperfusion in rabbits. AB - Reperfusion injury is a phenomenon complicating microvascular reconstruction. Extensive investigations of free-radical scavengers exist in the literature. The potential beneficial effects of verapamil, deferoxamine, and dimethylsulfoxide were studied in the rabbit forelimb subjected to 3 hr of tourniquet ischemia, followed by a period of reperfusion. Five minutes prior to tourniquet release, rabbits were given single intravenous infusions of normal saline, verapamil (0.2 mg/kg), deferoxamine (50 mg/kg), or dimethylsulfoxide (100 mg/kg). Following reperfusion, neither light microscopy nor laser Doppler flowmetry revealed significant differences between the sham and treated limbs. Muscle function was evaluated by measuring maximal twitch tensions in the flexor digitorum superficialis. Deterioration of muscle contractile function at 0.5, 3, and 24 hr after reperfusion was significantly improved by treatment with verapamil or deferoxamine. Conversely, dimethylsulfoxide worsened muscle function post ischemia. Dimethylsulfoxide does not appear to be beneficial, while verapamil and deferoxamine may be of benefit in mitigating reperfusion injury in microvascular transplantation. PMID- 8726340 TI - Ring avulsion injuries: microsurgical management. AB - Microsurgical treatment of ring avulsion injuries is often difficult, due to widespread intimal damage of the digital vascular bundles. Thirty-one patients with such injuries in varying degrees were treated during the period 1986 to 1992: of these, 15 underwent microsurgical revascularization. In addition to the traditional procedures applied in seven patients (arteriolysis, direct vessel suture, vein grafts), a technique consisting of vessel transfer from the middle finger was also employed. In five patients, the ulnar digital artery of the middle finger was transferred to the ring finger. In four patients, at least one vein was transferred from the dorsal aspect of the middle finger. Twelve of 15 microreconstructions were successful: with an average follow-up of 48.3 months, these patients showed very good functional (mean total active range of motion: 234 degrees and mean s2PD: 9.8mm) and cosmetic recovery. These results lead to the conclusion that, except for cases characterized by proximal amputation at the flexor superficialis tendon insertion, microsurgical treatment should always be carried out. PMID- 8726341 TI - Thrombolysis at a controlled pressure prolongs the survival of skin flaps treated with superoxide dismutase. AB - The role of thrombolysis in reestablishing patency in the microcirculation following ischemia, and thereby improving the efficacy of agents attenuating reperfusion injury, such as the oxygen free radical scavenger, superoxide dismutase (SOD), was investigated in a rat model. Abdominal skin flaps were subjected to normothermic ischemia induced by complete occlusion of the pedicle for periods of 12, 13, 14, 16, 18, 20, 22, and 24 hr. In Group 1 (n = 64), all animals received flap washout using 100,000U urokinase (manual injection) followed by 7,500 IU SOD given intra-arterially immediately prior to reperfusion. Animals in Group 2 received flap washout consisting of 100,000U urokinase given via a pressurized delivery system, followed by 7,500 IU SOD. Results demonstrated a statistically significant improvement in flap survival in Group 2. The authors concluded that thrombolytic therapy may be useful in improving the delivery of agents, such as SOD, which attenuate reperfusion injury in skin flaps. PMID- 8726342 TI - Cross-facial nerve grafting for facial reanimation: effect on normal hemiface motion. J Reconstr Microsurg 12:99, 1996. PMID- 8726343 TI - Anabolic steroid use and associated health risk behaviours. AB - Androgenic-anabolic steroid use is a significant problem among young people. The abuse of steroids may cause significant medical problems. Adolescent steroid users have also been found to be more likely than non-users to be engaging in other high risk behaviours including multiple drug use, cigarette and smokeless tobacco use, suicidal behaviours, high risk sexual behaviours, carrying a weapon, and fighting behaviours. It is critical that clinicians and athletic coaches and trainers are aware of these associations in order to more carefully screen and advise adolescents for health risk behaviours. PMID- 8726344 TI - Factors associated with respiratory problems in swimmers. PMID- 8726345 TI - Applied physiology of water-skiing. AB - Water-skiing is a physically demanding sport involving highly coordinated movements, extreme upper body torques, sustained isometric contractions and near maximal stresses on the anaerobic system. Due to the physical demands of the sport, elite skiers have developed physiological characteristics similar to other elite athletes. However, until recently little research has focused on the kinesiology, physiology and physical conditioning of water-skiing. PMID- 8726346 TI - The effects of acute and chronic exercise on sleep. A meta-analytic review. AB - Studies attempting to ascertain the effects of acute and chronic exercise on measures of sleep have yielded conflicting results and interpretations. Methodological differences among studies may explain this lack of consensus; however, small sample sizes and subsequently low statistical power may also have contributed. In an attempt to resolve these issues, this review used meta analytical techniques to: (a) re-examine the effects of exercise on sleep; and (b) examine possible moderators of these effects. Studies meeting the selection criteria were included in the analysis. Analyses of moderating factors were performed for stage 4 sleep and rapid eye movement (REM) sleep. The results indicated that acute and chronic exercise increased slow wave sleep (SWS) and total sleep time but decreased sleep onset latency and REM sleep. Moderating variables influencing the magnitude and direction of these effects were related to characteristics of the individual (e.g. sex, age, fitness level) and the exercise (e.g. time of day exercise was completed, type of exercise, exercise duration). Mechanisms which have been suggested to explain the relationship between exercise and sleep are discussed and directions for further research are provided. PMID- 8726347 TI - Circadian variation in sports performance. AB - Chronobiology is the science concerned with investigations of time-dependent changes in physiological variables. Circadian rhythms refer to variations that recur every 24 hours. Many physiological circadian rhythms at rest are endogenously controlled, and persist when an individual is isolated from environmental fluctuations. Unlike physiological variables, human performance cannot be monitored continuously in order to describe circadian rhythmicity. Experimental studies of the effect of circadian rhythms on performance need to be carefully designed in order to control for serial fatigue effects and to minimise disturbances in sleep. The detection of rhythmicity in performance variables is also highly influenced by the degree of test-retest repeatability of the measuring equipment. The majority of components of sports performance, e.g. flexibility, muscle strength, short term high power output, vary with time of day in a sinusoidal manner and peak in the early evening close to the daily maximum in body temperature. Psychological tests of short term memory, heart rate-based tests of physical fitness, and prolonged submaximal exercise performance carried out in hot conditions show peak times in the morning. Heart rate-based tests of work capacity appear to peak in the morning because the heart rate responses to exercise are minimal at this time of day. Post-lunch declines are evident with performance variables such as muscle strength, especially if measured frequently enough and sequentially within a 24-hour period to cause fatigue in individuals. More research work is needed to ascertain whether performance in tasks demanding fine motor control varies with time of day. Metabolic and respiratory rhythms are flattened when exercise becomes strenuous whilst the body temperature rhythm persists during maximal exercise. Higher work-rates are selected spontaneously in the early evening. At present, it is not known whether time of day influences the responses of a set training regimen (one in which the training stimulus does not vary with time of day) for endurance, strength, or the learning of motor skills. The normal circadian rhythms can be desynchronised following a flight across several time zones or a transfer to nocturnal work shifts. Although athletes show all the symptoms of 'jet lag' (increased fatigue, disturbed sleep and circadian rhythms), more research work is needed to identify the effects of transmeridian travel on the actual performances of elite sports competitors. Such investigations would need to be chronobiological, i.e. monitor performance at several times on several post-flight days, and take into account direction of travel, time of day of competition and the various performance components involved in a particular sport. Shiftwork interferes with participation in competitive sport, although there may be greater opportunities for shiftworkers to train in the hours of daylight for individual sports such as cycling and swimming. Studies should be conducted to ascertain whether shiftwork-mediated rhythm disturbances affect sports performance. Individual differences in performance rhythms are small but significant. Circadian rhythms are larger in amplitude in physically fit individuals than sedentary individuals. Athletes over 50 years of age tend to be higher in 'morningness', habitually scheduling relatively more training in the morning and selecting relatively higher work rates during exercise compared with young athletes. These differences should be recognised by practitioners concerned with organising the habitual regimens of athletes. PMID- 8726348 TI - Management of back pain in athletes. AB - Back pain affects millions of people. It affects 80% of the population and up to 52% at any given time. Back pain is not limited to sedentary individuals; it has significant effects on athletes as well. Depending upon the sport, incidence rates of back pain occur in athletes from 1.1% to as high as 30%. Athletes differ from the non-athletic population in that their incentives to return to activity are considerably different than non-athletes. The reasons may vary from the will to win through to significant financial considerations. Although reasons for recovery are different, the physiology and mechanics of repair of injured soft tissue in the athlete is the same as for the non-athlete. Proper management of the athlete requires ruling out emergent causes of back pain such as tumour, infection, acute fracture, progressive neurological deficit, visceral sources (e.g. pancreatitis, abdominal aortic aneurysm), and rheumatoid variants. Once a good history and physical is performed, a simple classification system can be utilised to manage the athlete presenting with back pain. This system can be expressed as: (a) regional back pain; (b) radicular leg pain; (c) radicular leg pain with progressive neurological deficit; and (d) cauda equina syndrome. Each of these categories needs to be managed in a specific manner and can provide the healthcare professional with simple, straightforward guidelines for handling the athlete with lower back pain. The key is to return the athlete to the field of play in a safe and timely manner. PMID- 8726349 TI - Homologous recombination in variants of the B16 murine melanoma with reference to their metastatic potential. AB - Genomic instability has been accepted as providing a phenotypic variety of malignant cells within a developing tumour. Defects in genetic recombination can often lead to phenotypic differences; therefore, it is possible that metastatic variant cell lines exhibit their particular phenotype as a result of an altered ability to catalyse homologous recombination. We have investigated recombination efficiency in B16 melanoma metastatic variants, using a plasmid, pDR, as a recombination substrate. The plasmid contains two truncated, nontandem but overlapping segments of the neomycin resistance gene (neo 1 and neo 2), separated by the functional gpt gene unit. Only a successful recombination of the two neo segments will generate a functionally intact neomycin gene. Extrachromosomal recombination here was a transient measure of the cells to recombine the neo fragments in an intra- or intermolecular manner. Extrachromosomal recombination frequencies were higher in the high metastasis variants (BL6, ML8) compared with the low metastatic F1 cells. On the other hand, the frequency of chromosomal recombination (after plasmid integration) was higher for the low metastasis (F1) cell line compared with the highly metastatic variants, BL6 and ML8. Since the recombination assay measures only successful recombination events, we have interpreted the observed higher incidence of chromosomal recombination in the low metastatic variant line as indicative of a more stable genome. Similarly, a higher inherent instability in the genome of the high metastasis variants would render these less efficient at producing and maintaining successful recombination events, and this was found to be true by Southern analysis. The results presented show that frequency of recombination may be adduced as evidence for implicating genomic instability in the generation of variant cell populations during metastatic spread. Such an interpretation is also compatible with the Nowell hypothesis for tumour progression. PMID- 8726350 TI - Tamoxifen induces TGF-beta 1 activity and apoptosis of human MCF-7 breast cancer cells in vitro. AB - We report here that the antiestrogen tamoxifen (TAM) induces cell death in human breast cancer cell line MCF-7. We assessed the type of cell death induced by TAM in this breast cancer cell line on the basis of morphological and biochemical characteristics. Dying cells showed morphological characteristics of apoptosis, such as chromatin condensation and nuclear disintegration. DNA isolated from these cells revealed a pattern of distinctive DNA bands on agarose gel. The DNA fragmentation in MCF-7 cells induced by TAM could also be detected by terminal deoxynucleotidyl transferase-mediated dUTP-biotin end labeling. Northern blot hybridization revealed a substantial increase in the amounts of TRPM-2 and TGF beta 1 mRNAs in MCF-7 cells after treatment with TAM. In contrast, the mRNA level of the estrogen-induced pS2 gene was strongly suppressed. The biological activity of TGF-beta was increased at least fourfold in the media from MCF-7 cells treated with TAM. The results presented in this study suggest that TAM induces apoptosis of MCF-7 cells and it may be mediated by the secretion of active TGF-beta. PMID- 8726351 TI - Prostate-derived soluble factors block osteoblast differentiation in culture. AB - Bone metastasis is a common event and a major cause of morbidity in prostate cancer patients. After colonization of bone, prostate cells induce an osteoblastic reaction which is not associated with marrow fibrosis (i.e., osteoblast but not fibroblast proliferation). In the present study we test the hypothesis that the tumoral prostatic cell line (PC-3) secretes factors that block the osteoblast differentiation process, resulting in an increase of the relative size of the proliferative cell pool. Our results, using fetal rat calvaria cells in culture, show that conditioned medium from PC-3 cells (PC-3 CM) stimulates osteoblast proliferation and inhibits both alkaline phosphatase (AP) activity (an early differentiation marker) and the mineralization process, measured as calcium accumulation (late differentiation marker). The inhibition of the expression of AP and mineralization depends on the presence of PC-3 CM during the proliferative phase of culture and suggests that both processes occur in a nonsimultaneous fashion. The inhibitory effect of PC-3 CM was not reverted by dexamethasone, which would indicate that prostatic-derived factors and the glucocorticoid do not share a common site of action. Measurement of the proliferative capacity of subcultures from control and treated cells demonstrates that PC-3 CM treatment induces the maintenance of the proliferative potential that characterizes undifferentiated precursor cells. PMID- 8726352 TI - Identification of a ubiquitin-like protein in the mammalian vitreous humor. AB - An 8 kDa ubiquitin-like peptide (ULP) was isolated by high performance liquid chromatography from the rabbit vitreous humor, and the N-terminal amino acid sequence of this peptide showed complete homology with ubiquitin. Western blot revealed the presence of free ULP in both the iris-ciliary (IC) complex and the aqueous humor extracts. In the IC complex, fluorescence and immunoelectron microscopy detected high concentrations of ULP in the posterior epithelial cells, suggesting this tissue as a possible source of ULP in the ocular fluids. Significantly, this is the first time that the presence of free ULP has been reported in mammalian extracellular fluids. Furthermore, we recently demonstrated that the 8 kDa fraction of vitreous humor containing ULP is a potent inhibitor of protein synthesis [Banerjee et al. (1992): J Cell Biochem 49:66-73]. These findings taken together suggest a novel biological role for ULP in the control of lens cell growth. PMID- 8726353 TI - Cell density-dependent changes in the insulin action pathway: evidence for involvement of protein-tyrosine phosphatases. AB - In order to examine alterations in the phosphorylation state of proteins involved in insulin action that might accompany the reduced growth state of density arrested cells, we measured the insulin-stimulated phosphorylation of the receptor and high M(r) cellular substrates of the receptor kinase in rat hepatoma cells at different cell densities. As cell density increased from 2 x 10(5) to 3.2 x 10(6) per 35-mm well, the rate of DNA synthesis fell to 22% of control, while insulin-stimulated tyrosine phosphorylation of high M(r) receptor substrates ("pp185") was enhanced to 198% of control, without a change in the abundance of insulin receptor substrate (IRS)-1 protein. In anti-IRS-1 immunoprecipitates, tyrosine phosphorylation was increased by only 30%, suggesting that increased tyrosine phosphorylation of additional high M(r) proteins (e.g., IRS-2) accounted for much of the observed increase in tyrosine phosphorylation of the receptor substrates. In spite of increased tyrosine phosphorylation of IRS-1 and total pp185-related proteins, however, cells studied at high growth density exhibited a 25% decrease in IRS-1-associated phosphatidylinositol 3'-kinase activity and only a 39% increase in phosphatidylinositol 3'-kinase activity in antiphosphotyrosine immunoprecipitates. To explore the potential role of hepatic protein-tyrosine phosphatases (PTPases) in the hyperphosphorylation of pp185 proteins, we found by immunoblotting that at high cell density the intracellular PTPase PTP1B and the transmembrane PTPase LAR were reduced in abundance by 49% and 55%, respectively, while the abundance of the SH2-domain containing PTPase SH-PTP2 was increased by 48%. These data demonstrate that the attenuation of post-receptor signaling by insulin in hepatoma cells at increasing growth density involves changes in endogenous substrate phosphorylation which may result from alterations in specific PTPases implicated in the regulation of the insulin action pathway. PMID- 8726354 TI - Ligation of the alpha 2-macroglobulin signaling receptor on macrophages induces synthesis of platelet activating factor. AB - The binding of receptor-recognized forms of alpha 2-macroglobulin (alpha 2M) to macrophage alpha 2M signaling receptors increases inositol-1,4,5-triphosphate synthesis and induces Ca2+ mobilization. In this report, we demonstrate that ligation of the macrophage alpha 2M signaling receptor is also associated with synthesis of platelet activating factor (PAF) by both the de novo and remodeling pathways. Both alpha 2M-methylamine and a cloned and expressed 20-kDa receptor binding fragment (RBF) from rat alpha 1M+, stimulated macrophage synthesis of PAF from [3H]acetate, [3H]methylcholine, and 1-O-[3H]alkyl lyso-PAF by two- to threefold. PAF levels reached a peak in 20 min after the cells were exposed to alpha 2M-methylamine or RBF; they remained elevated for about 1 h after ligand addition to the cells. When [3H]methylcholine was the substrate, pertussis toxin did not block PAF synthesis, but the protein kinase C inhibitor staurosporin reduced synthesis by 65-70%. Cycloheximide completely abolished the increase in synthesis of PAF by macrophages exposed to alpha 2M-methylamine. By contrast, when [3H]acetate was employed as a precursor, staurosporin or cycloheximide did not abolish the increase in PAF synthesis. These studies suggest that protein kinase C is necessary for the induction of the de novo pathway by alpha 2M methylamine. Both alpha 2M-methylamine and RBF stimulated the activity of lyso PAF acetyltransferase by about fourfold. Both ligands also stimulated the activity of PAF acetylhydrolase by about six- to sevenfold, indicating that ligation of the alpha 2M signaling receptor also regulates the degradation of PAF. The ability of receptor-recognized forms of alpha 2M to regulate levels of PAF suggests that alpha 2M-proteinase complexes not only regulate macrophage function by activating intracellular signaling but also may indirectly regulate the function of other cells that cannot bind alpha 2M-proteinase complexes. PMID- 8726355 TI - Product of the oncogene-activating gene Tpr is a phosphorylated protein of the nuclear pore complex. AB - We have identified a component of the human nuclear pore complex and have shown that it is the product of a gene involved in oncogenic activation. A monoclonal antibody raised against purified nuclear matrix proteins recognizes a single protein with an electrophoretic mobility of approximately 300 kDa and stains the nuclear envelope in a punctate pattern typical of nuclear pores. The antibody was used to screen lambda gt11 human cDNA libraries, and the resulting clones were sequenced and compared to sequences in the Genbank database. An exact match was found with the human tpr (for translocated promoter region) gene, a gene shown previously to be involved in the oncogenic activation of several protein kinases. Double-label immunofluorescent microscopy with the anti-Tpr antibody and an antibody to the previously characterized nuclear pore complex protein nup153 confirms that Tpr is localized to the nuclear pore complex. Tpr is located on the cytoplasmic face of the nucleus, as demonstrated by immunofluorescent staining of cells permeabilized with digitonin. Tpr is a 2,349-amino acid protein with extensive coiled-coil domains and an acidic globular C-terminus. The protein contains 10 leucine zipper motifs and numerous sites for phosphorylation by a variety of protein kinases. Immunoprecipitation of Tpr from 32P-orthophosphate labeled cells shows that it is a phosphoprotein. Potential functions for Tpr and possible mechanisms for the transforming activity of Tpr fusion proteins are discussed. PMID- 8726356 TI - Binding of rat alpha 1-inhibitor-3-methylamine to the alpha 2-macroglobulin signaling receptor induces second messengers. AB - Binding of receptor-recognized forms of tetrameric human alpha 2-macroglobulin (alpha 2M*) to a macrophage signaling receptor induces cAMP synthesis, increases in inositol 1,4,5-triphosphate (IP3) synthesis, and a concomitant rise in cytosolic free calcium ([Ca2+]i). The alpha 2M* signaling receptor is coupled to a pertussis-toxin insensitive G protein. Binding of alpha 2M* also occurs to the low density lipoprotein receptor-related protein/alpha 2M receptor (LRP/alpha 2MR), but this binding does not induce signal transduction. Rat alpha 1-inhibitor 3 (alpha 1I3) is a monomeric member of the alpha-macroglobulin/complement superfamily. Like alpha 2M, it can react with proteinases or methylamine which induces a conformational change causing activated alpha 1I3 to bind to LRP/alpha 2MR. We now report that alpha 1I3-methylamine binds to the macrophage alpha 2M* signaling receptor inducing a rapid rise in the synthesis of IP3 with a subsequent 1.5- to 3-fold rise in [Ca2+]i. alpha 1I3-methylamine binding to macrophages also caused a statistically significant elevation in cAMP. Native alpha 1I3, like alpha 2M, was unable to induce signal transduction. alpha 1I3 forms a complex with alpha 1-microglobulin, which has a distinct conformation from alpha 1I3 and is recognized by LRP/alpha 2MR. This complex also induces an increase in [Ca2+]i comparable to the effect of alpha 1I3-methylamine on macrophages. It is concluded that activation of alpha 1I3 by methylamine or binding of alpha 1-microglobulin causes similar conformational changes in the inhibitor, exposing the receptor recognition site for the alpha 2M* signaling receptor, as well as for LRP/alpha 2MR. PMID- 8726357 TI - Effect of hypoxia on hepatic DNA methylation and tRNA methyltransferase in rat: similarities to effects of methyl-deficient diets. AB - Young rats were maintained in a 10% oxygen atmosphere for 2, 6, and 10 days and administered normal rat chow and water ad libitum. Thereafter, their hepatic S adenosyl-L-methionine (AdoMet) and activity and mRNA levels of AdoMet synthetase were assayed. AdoMet levels decreased by 45% after 10 days; hepatic AdoMet synthetase also declined by approximately 40%. In rats with low hepatic AdoMet, the mRNA level of AdoMet synthetase also declined by up to 80%. No significant change in AdoMet or AdoMet synthetase was noted in pair-fed normoxic rats. DNA hypomethylation was determined in terms of incorporation of [3H]methyl of AdoMet incorporated at unmethylated sites in DNA in reactions mediated by methylases HpaII and SssI. As compared to the normal hepatic DNA, [3H]methyl group incorporation in the 10-day hypoxic DNA was almost double in the HpaII-mediated reaction and approximately 10-fold in the SssI-mediated reaction. Hepatic tRNA methyltransferase activity doubled after 10 days of hypoxia. However, hypoxic rats showed no detectable mRNA transcripts for c-myc and c-fos oncogenes on Northern blot analysis. These observations show that because of subnormal activity of AdoMet synthetase, hypoxic liver is depleted of AdoMet, even when the animals are administered a complete diet. However, unlike rats on chronic lipotrope-deficient diets, hypoxic rats on a complete diet show no aberrant expression of oncogenes. PMID- 8726358 TI - Intracellular localization of the mRNAs of argininosuccinate synthetase and argininosuccinate lyase around liver mitochondria, visualized by high-resolution in situ reverse transcription-polymerase chain reaction. AB - Argininosuccinate synthetase and argininosuccinate lyase, two cytoplasmic enzymes of the urea cycle, are released into the soluble phase in the absence of detergent when cells are disrupted. Yet previous biochemical studies, as well as immunocytochemistry at the electron microscope level, have shown that these enzymes are localized around mitochondria in situ. Such intracellular localization of soluble enzymes requires mechanisms to deliver the proteins to the appropriate sites, where they may then be anchored by specific protein protein interactions. A method was developed to examine the intracellular distribution of the mRNA of argininosuccinate synthetase and argininosuccinate lyase in intact rat liver at the ultrastructural level by in situ reverse transcription and the polymerase chain reaction, using primers targeting regions of the coding sequences of the rat enzymes, digoxigenin-dUTP as the label, and anti-digoxigenin/1 nm [corrected] gold plus silver enhancement as the detection method. The tissue was fixed in 4% paraformaldehyde/0.1% glutaraldehyde and embedded in Lowicryl. Examination of the numbers and the location of the silver grains, coupled with morphometric analysis of the electron micrographs, permitted the calculation of the silver "enrichment ratio" for each type of cell structure. These ratios showed that the mRNAs for argininosuccinate synthetase and argininosuccinate lyase were located next to the cytoplasmic side of the mitochondrial membrane and in the nearby endoplasmic reticulum. Most of the silver grains that were observed in the endoplasmic reticulum were within 200 nm of the mitochondria; it was not possible, however, to determine if those grains were actually associated with the reticular membranes. These studies demonstrate that the mRNAs of these two soluble cytoplasmic proteins are localized to the same limited regions where the proteins are situated. Translation of the proteins, therefore, must occur at these specific sites. The targeting of argininosuccinate synthetase and argininosuccinate lyase mRNAs to the immediate vicinity of the mitochondria may be the first step of the mechanisms by which the spatial organization of these soluble proteins in situ is accomplished. The targeting of mRNAs for soluble cytoplasmic proteins of organized metabolic pathways has not been demonstrated previously. These studies also show that in situ reverse transcription and the polymerase chain reaction at the ultrastructural level, which has not been previously reported, can be used to detect specific mRNAs; it should be extremely valuable for the intracellular detection of low-abundance mRNAs. PMID- 8726359 TI - Inhibition of ADP-induced platelet activation by 7-chloro-4-nitrobenz-2-oxa-1,3 diazole: covalent modification of aggregin, a putative ADP receptor. AB - ADP-induced platelet responses play an important role in the maintenance of hemostasis. There has been disagreement concerning the identity of an ADP receptor on the platelet surface. The chemical structure of 7-chloro-4-nitrobenz 2-oxa-1,3-diazole (NBD-CI) shows considerable resemblance to that of the adenine moiety of adenine-based nucleotides. The reagent has been previously used by other investigators as an affinity label for adenine nucleotide-requiring enzymes, such as mitochondrial ATPase and the catalytic subunit of cAMP-dependent protein kinase. Since ADP-induced platelet responses depend on the binding of ADP to its receptor, we investigated the effect on ADP-induced platelet responses and the nature of ADP-binding protein modified by NBD-CI. NBD-CI inhibited ADP induced shape change and aggregation of platelets in platelet-rich plasma in a concentration- and time-dependent manner. NBD-CI also inhibited ADP-induced shape change, aggregation, exposure of fibrinogen binding sites, secretion, and calcium mobilization in washed platelets. NBD-CI did not act as an agonist for platelet shape change and aggregation. Covalent modification of platelets by NBD-CI blocked the ability of ADP to antagonize the increase in intracellular levels of cAMP mediated by iloprost (a stable analogue of prostaglandin I2). NBD-CI was quite specific in inhibiting platelet aggregation by those agonists, e.g., ADP, collagen, and U44619 (a thromboxane mimetic), that completely or partially depend on the binding of ADP to its receptor. Autoradiogram of the gel obtained by SDS PAGE of solubilized platelets modified by [14C]-NBD-CI showed the presence of a predominant radiolabeled protein band at 100 kDa corresponding to aggregin, a putative ADP receptor. The intensity of this band was considerably decreased when platelets were either preincubated with ADP and ATP or covalently modified by a sulfhydryl group modifying reagent before modification by [14C]-NBD-CI. These results (1) indicate that covalent modification of aggregin by NBD-CI contributed to loss of the ADP-induced platelet responses, and (2) suggest that there is a sulfhydryl group in the ADP-binding domain of aggregin. PMID- 8726360 TI - Decreased heterogeneity of CS histone variants after hydrolysis of the ADP-ribose moiety. AB - Sea urchin CS histone variants are electrophoretically heterogeneous when analyzed in two dimensional polyacrylamide gels (2D-PAGE). Previous results suggested that this heterogeneity is due to the poly (ADP-ribosylation) of these proteins. Consequently, native CS histone variants were subjected to different treatments to remove the ADP-ribose moiety. The incubation in 1 M hydroxylamine was not effective in eliminating the polymers of ADP-ribose from CS variants, and the treatment with sodium hydroxide was deleterious to the proteins. In contrast, the ADP-ribose moiety was successfully removed from the CS variants by incubation with phosphodiesterase (PDE). To eliminate contamination of CS histone variants with PDE extract, the enzyme was covalently bound to Sepharose 4B prior to its utilization. Treatment of native CS histone variants with this immobilized phosphodiesterase removed around 85% of the total ADP-ribose moiety from these proteins. After S-PDE treatment the complex electrophoretic pattern of CS histone variants in 2-D PAGE decreases to five major fractions. From these results we conclude that the electrophoretic heterogeneity of native CS histone variants is mainly due to the extent to which five main CS histone variants are poly(ADP) ribosylated). PMID- 8726361 TI - Influence of subunit transcript and protein levels on formation of a mitochondrial multienzyme complex. AB - Constitutive expression of nuclear genes encoding mitochondrial proteins raises the question of whether these proteins are present in similar amounts in mitochondria of different tissues. We report that amounts of a single multienzyme complex can vary on a per mitochondrion basis depending on the number of mitochondria per cell. Human branched-chain alpha-keto acid dehydrogenase (BCKD) expression is used as a paradigm in these studies. Expression is compared and contrasted in HepG2 and DG75 cells in which mitochondrial content is twofold higher in the hepatocarcinoma line than in the lymphoblastoid line. Per cell, BCKD activity is equal in the two cells types, but BCKD protein concentration per mitochondrion is twofold higher in DG75 cells. Steady-state mRNA levels do not appear to be directly related to amounts of protein in the two cell lines. To test whether one subunit is limiting in formation of complex, overexpression of each BCKD subunit was elicited by plasmid transfection of the DG75 cells. Only overexpression of the beta-subunit of the decarboxylase component induced more BCKD activity without apparent increase in mRNA for the other endogenously expressed subunits. This implies that free BCKD subunits exist in a cell and can be recruited into an active complex when the limiting subunit becomes available. PMID- 8726362 TI - Induction of mouse beta integrin expression following transfection with human alpha 4 chain. AB - We report here an analysis of the expression and function of the alpha chain of human VLA-4 in stable mouse L cell transfectants and the requirement for the beta chain in these processes. L cells were transfected with human alpha 4 cDNA or alpha 4 and human beta 1 cDNA. Unexpectedly, human alpha 4 cDNA, when transfected alone, could induce de novo surface expression of host beta 7 and increased expression of host beta 1. Induction of mouse beta 7 and beta 1 surface expression was not due to de novo gene activation, but instead represented alpha 4/beta intracellular subunit association and transport to the cell surface. Transfection with human beta 1 prevented surface expression of mouse beta integrins. Whereas human alpha 4 and human beta 1 subunits associated very tightly in anti-alpha 4 immunoprecipitates, human alpha 4 and mouse beta subunits were only partially associated. Furthermore, binding of human/mouse chimeric receptors to recombinant VCAM, a major ligand for alpha 4 beta 7 and alpha 4 beta 1, was very poor, whereas human alpha 4/human beta 1 receptors bound strongly to VCAM. One alpha 4 transfectant, which exhibited a tight human alpha 4/mouse beta 1 association, could be induced, but only after PMA activation, to bind strongly to VCAM. These results indicate that alpha 4 subunits have specific affinity for beta 7 and beta 1 integrins and require beta subunits for surface expression as well as high affinity ligand binding activity. Our results indicate that a tight association between the alpha 4 and beta subunit appears to be critical for ligand binding, consistent with a direct as well as regulatory role for the beta subunit in ligand binding. Furthermore, these studies demonstrate that expression of foreign recombinant proteins can alter host cell protein expression resulting in de novo surface protein expression. PMID- 8726363 TI - Reduction-oxidation (redox) state regulation of extracellular matrix metalloproteinases and tissue inhibitors in cardiac normal and transformed fibroblast cells. AB - Latent matrix metalloproteinases (MMPs) in normal myocardium are activated in end stage heart failure. In vitro oxidized glutathione (GSSG) activates myocardial MMPs which contains a cysteine residue. In vivo GSSG induce the collagen lysis and cardiac dilatation. To assess whether thiol and non-thiol reducing agents have direct effect on the interstitial human heart fibroblast (HHF) proliferation and MMP expression, HHF and polyoma virus transformed fibroblast cells were cultured with or without the thiol-containing reduced (GSH) or oxidized (GSSG) glutathiones, pyrrolidine dithiocarbamate (PDTC) and N-acetylcysteine (NAC), and non-thiol ascorbic acid. After 100 micrograms/ml (approximately 0.3 mM) GSH or PDTC treatment the proliferative (synthetic) phenotype of transformed fibroblast cells was changed to quiescent (contractile) phenotype. Also, after GSH, PDTC, and ascorbic acid treatment the medium was then analyzed for MMP activity by zymography. The results indicate reduction in MMP expression in transformed fibroblast cells after GSH and PDTC treatments and no effect after ascorbic acid treatment. Based on reverse zymography, we observed the level of tissue inhibitor of metalloproteinase (TIMP) at a decreased level in transformed cells. The effect of the reducing agent at the gene transcription was measured by estimating mRNA (Northern blot analysis) of MMP and of TIMP in the cells that were cultured in medium in the presence and absence of GSH. These results indicate that GSH induces MMP-2 and MMP-1 expression in normal HHF and that GSH reduces MMP-2 and MMP-1 in transformed fibroblast cells. After the treatment, the TIMP-2 level was repressed in normal HHF and TIMP-2 level increased in transformed fibroblast cells. These events are dependent on the nuclear transcription factor activity on the collagenase promoter in normal HHF cells. On the other hand, in polyoma transform fibroblast cells these events are not dependent on this collagenase promoter. These results suggest that oxidative environment induces normal HHF cell proliferation, and the reducing agent decreases normal HHF cell proliferation by inducing MMP and repressing TIMP gene transcription. In transformed cells reducing agents inhibit MMP expression and increase TIMP levels, which suggests a role of antioxidants in preventing tumorigenesis. PMID- 8726365 TI - Speed of information processing as a mediator between age and free-recall performance. AB - A combined experimental and individual differences approach was used to investigate the mediating role of task-specific and task-independent speed of information processing measures in the relationship between age and free-recall performance. Thirty-six younger adults (mean age = 21 years) and 36 older adults (mean age = 73 years) participated. Participants were required to encode 3 lists of words for immediate recall, by rehearsing the words aloud, twice, and 3 times. Participants' speed of information processing was assessed by 3 measures: rehearsal time, articulation speed, and scores on the Digit Symbol Substitution Test (DSST; Wechsler, 1981). Working memory was also assessed by a backward word span measure. As predicted, younger adults recalled more words after rehearsing words 3 times rather than once, whereas older adults' recall did not increase with increasing numbers of rehearsals. Younger adults were faster on all speed-of processing measures and had higher backward word span than did older adults. Task independent speed of processing, measured by DSST scores and articulation speed, mediated the relationship between age and free recall. Scores on the DSST appear to reflect a fundamental difference between younger and older adults that influences recall performance. PMID- 8726364 TI - Acidic fibroblast growth factor inhibits osteoblast differentiation in vitro: altered expression of collagenase, cell growth-related, and mineralization associated genes. AB - Fibroblast growth factors (FGF) are osteoblast mitogens, but their effects on bone formation are not clearly understood. Most in vitro studies examining the effects of FGFs on osteoblasts have been performed only during the initial proliferative stage of osteoblast culture. In these studies, we examined the consequential effect of acidic FGF in cultures of rat fetal diploid osteoblasts that undergo a developmental differentiation program producing a mineralized bone like matrix. During the initial growth period (days 1-10), addition of acidic FGF (100 micrograms/ml) to actively proliferating cells increased (P < 0.05) 3H thymidine uptake (2,515 +/- 137, mean +/- SEM vs. 5,884 +/- 818 cpm/10(4) cells). During the second stage of maturation (days 10-15), osteoblasts form multilayered nodules of cells and accumulate matrix, followed by mineralization (stage 3, days 16-29). Addition of acidic FGF to the osteoblast cultures from days 7 to 15 completely blocked nodule formation. Furthermore, addition of acidic FGF after nodule formation (days 14-29) inhibited matrix mineralization, which was associated with a marked increase in collagenase gene expression, and resulted in a progressive change in the morphology of the nodules, with only a few remnants of nonmineralized nodules present by day 29. Histochemical and biochemical analyses revealed a decrease in alkaline phosphatase and mineral content, confirming the acidic FGF-induced inhibition of nodule and matrix formation. To identify mechanisms contributing to these changes, we examined expression of cell growth and bone phenotypic markers. Addition of acidic FGF during the proliferative phase (days 7-8) enhanced histone H4, osteopontin, type I collagen, and TGF-beta mRNA levels, which are coupled to proliferating osteoblasts, and blocked the normal developmental increase in alkaline phosphatase and osteocalcin gene expression and calcium accumulation. Addition of acidic FGF to the cultures during matrix maturation (days 14-15) reactivated H4, osteopontin, type I collagen, and TGF-beta gene expression, and decreased alkaline phosphatase and osteocalcin gene expression. In an in vivo experiment, rats were treated with up to 60 micrograms/kg/day acidic FGF intravenously for 30 days. Proliferation of osteoblasts and deposition of bone occurred in the marrow space of the diaphysis of the femur in a dose-related fashion. The metaphyseal areas were unaffected by treatment. In conclusion, our data suggest that acidic FGF is a potent mitogen for early stage osteoblasts which leads to modifications in the formation of the extracellular matrix; increases in TGF-beta and collagenase are functionally implicated in abrogating competency for nodule formation. Persistence of proliferation prevented expression of alkaline phosphatase and osteocalcin, also contributing to the block in the progression of the osteoblast developmental sequence. PMID- 8726366 TI - Search for semantic inhibition failure during sentence comprehension by younger and older adults. AB - The present research examined recent evidence that inhibition failures do not contribute to age-related declines in on-line text comprehension. Evaluating naming performance over a time course revealed that processes serving to eliminate thematically irrelevant information from memory in young adults were apparently deficient for older adults. Experiment 2 generalized these findings to contexts that rely on later information to disambiguate ambiguous words. Under these conditions, older adults were as able as younger adults to use context to disambiguate words. Despite the apparent disagreement between the experimental outcomes, it is argued that the present studies together are consistent and support the view that inhibition failures unlikely contribute to age differences in on-line sentence comprehension (at least) when the contexts are semantically constraining (i.e., self-contained). It is proposed that the detection of age related inhibition failures may depend on the extent to which adults must rely on internally generated processing constraints. PMID- 8726367 TI - Antecedents and contexts of generativity motivation at midlife. AB - A semiprojective measure of generativity motivation--the desire to contribute to the welfare of society--was developed and applied to a longitudinal sample of educated women. In support of previous theory that generativity represents a fusion of agency and communion, generativity at midlife was related to a combination of adolescent scores on achievement, affiliation, and power motivation. The measure was also related to independent indexes of generative wishes and Q-sort generativity. The importance of contextualizing generativity motivation is demonstrated: Generative women with careers found gratification through work, whereas generative women not working in careers experienced gratification through parenting. Generativity was also related to political consciousness. Finally, as young adults, generative women expressed gratitude toward mentors. The authors discuss theoretical implications for E. H. Erikson's (1950) concept of psychosocial generativity. PMID- 8726368 TI - Social support as a mediator in the relation between functional status and quality of life in older adults. AB - The relations among physical functioning, social support, depressive symptoms, and life satisfaction were examined in a national sample of 4,734 adults age 65 and older. Regression analyses were used to examine the relative importance of objective and subjective support measures in understanding the relation between physical impairment and quality of life. Impairment was associated with fewer friendship contacts, fewer family contacts, less perceived belonging support, and less perceived tangible aid, but only measures of perceived support predicted depressive symptomatology. A structural equation modeling approach was then used to explore the mediational role of perceived social support in the relation between impairment and quality of life variables. Results are consistent with the hypothesis that lower reported social support is an important reason for decreases in life satisfaction and increases in depressive symptoms found among older adult populations. Implications for understanding the role of social support in attenuating the effects of physical disability in older adults are discussed. PMID- 8726369 TI - Long-term cued recall of tasks in senile dementia. AB - Participants with senile dementia of the Alzheimer's type, vascular dementia, or both, associated a task with a cue. On reinstatement of the cue 1 day later, a substantial portion of the sample recalled the task. The teaching method, both with and without participant performance of the task (PPT), was spaced retrieval with supplementary or fading cues provided as required. Findings were that (a) PPT encoding and retrieval encoding, separately, assisted later recall: (b) retrieval combined with PPT encoding increased the probability of task performance at final recall; (c) repetition in the absence of retrieval or PPT was less effective; and (d) there was no forgetting between 1 hr and 1 day. Theoretical and clinical aspects of these findings are discussed. PMID- 8726370 TI - Patterns of personality change across the life span. AB - Six aspects of personality were examined for a sample of 211 individuals across either a 30- or 40-year time span. Latent curve analyses found an underlying pattern of lifetime change for self-confidence, cognitive commitment, outgoingness, and dependability. An underlying pattern of lifetime stability was found for assertiveness. No shared systematic underlying pattern could be fit to individuals' lifetime trajectories for warmth. Although individuals shared a single underlying pattern of change or stability for 5 of the 6 dimensions of personality considered here, they also showed differences in the degree and direction of personality change. PMID- 8726371 TI - Moral and social reasoning and perspective taking in later life: a longitudinal study. AB - In this study 27 older adults (ages 64-80) and 23 middle-aged adults (ages 35-54) were tested for moral stage, integrative complexity of social reasoning, and perspective-taking levels twice over a 4-year period. Moral reasoning stage levels did not change over time for either age group. Older adults, but not the middle-aged, showed a significant decline over time in level of moral perspective taking. Complexity of reasoning about several interpersonal social issues declined modestly in both age groups. More social-cognitive support, a higher education level, and better self-reported health were all found to be protective factors in forestalling declines in mature adults' sociocognitive reasoning, consistent with other research on cognitive measures in later life. PMID- 8726372 TI - Age-related impairment in an event-based prospective-memory task. AB - Slides of famous people were presented to participants with the instructions to name each face and circle the trial number if the person was wearing glasses (prospective-memory target event). Participants in their 50s and 60s (n = 56) were more successful than participants in their 70s and 80s (n = 59) at both the naming an prospective-memory tasks. An age-related increase in the probability of forgetting replicated an earlier prospective-memory study (E. A. Maylor, 1993); in the present case, there was also an age-related decrease in the probability of recovery. These effects of age remained significant after other measures of current ability were taken into account, including intelligence, speed, and naming performance. For participants who were in both the earlier study (E. A. Maylor, 1993) and this study (n = 65), the correlation between prospective-memory performance on the 2 occasions was significant but only for younger participants. Performance in the prospective-memory task was entirely unrelated to performance in the naming task. PMID- 8726373 TI - Temporal discounting in choice between delayed rewards: the role of age and income. AB - This study examined the effects of age and income temporal discounting (i.e. the decrease in the subjective value of a reward as the delay to its receipt increases). The value of delayed hypothetical monetary rewards was discounted at similar rates by adults of different ages but similar income levels, but at different rates by adults of similar age but different income levels. Specifically, lower income older adults showed a greater degree of temporal discounting than did either upper income older adults or upper income younger adults, but there were no age differences in discounting between the upper income groups. Comparison of these findings with those of a previous study (Green, Fry, & Myerson, 1994) suggests that impulsivity in decision making declines rapidly in young adulthood, reaching stable levels in the 30s. Further, age and income appear to interact in determining the impulsivity of decision making by adults. PMID- 8726374 TI - Things that go bump in your life: explaining the reminiscence bump in autobiographical memory. AB - Two experiments explore the reminiscence bump (RB)--the disproportionately higher recall of early-life memories--by older adults. In Experiment 1, participants in the age ranges of 36-40, 46-50, and 56-60 recalled events freely or under instructions to avoid recent memories. Constraint did not affect older participants but resulted in the appearance of an RB in younger participants. In Experiment 2, recall was constrained to particular life periods. Memories from these periods were compared for ease of retrieval and along subjective dimensions (e.g. vividness). Memories from early life were more easily retrieved, but this was not due to differences in subjective qualities. A higher proportion of memories for first-time events were identified from early life, and these memories were more easily retrievable. The results are discussed in relation to an existing model of autobiographical memory, and a revised model is put forward. PMID- 8726375 TI - Does job-related training performance decline with age? AB - The meta-analytic review presents the findings of a project investigating the relationship between age and job-related training outcomes. The analysis is based on 83 effect sizes derived from 6,610 individuals. Results showed poorer training performance for older adults. In general, older adults, relative to younger adults, showed less mastery of training material (r = -.26), completed the final training task more slowly (r = .28), and took longer to complete the training program (r = .42). Field samples generally showed smaller age effects than laboratory samples. Estimated training performance for average individuals at various ages is provided. PMID- 8726376 TI - Effects of age on state of awareness following implicit and explicit word association tasks. AB - Younger and older participants did word-association tasks after implicit and explicit instructions and a read-generate study manipulation. No age differences were shown in the implicit version of the test. A generation effect for both age groups suggested that word-association priming can be classified as a conceptually driven task and a new task at which older adults show a relatively preserved memory function. However, the younger group did better on the explicit test in the generate condition. Participants were asked to examine their implicitly produced responses to make them accessible to conscious retrieval. Remember (R) and Know (K) measures of conscious awareness were applied to both postimplicit and postexplicit word-association responses. Age and awareness showed opposite effects in postimplicit retrieval. Younger participants tended to make more R responses than did the older adults, and K responses did not vary with age, but the older group was unaware of more primed items as study list members. Age differences were also shown in R but not K responses after word association cued recall. PMID- 8726377 TI - Depressive symptom experiences among older women: a comparison of alternative measurement approaches. AB - Age differences in depressive symptom experiences were investigated in a community sample of women (N = 287, ages 55-95 years). The purpose of the analysis was to compare the age-depression relation using 3 different measurement approaches: (a) a standard screening scale, (b) a clinical diagnostic, and (c) a latent variable measurement approach. The analysis, which uses structural equation modeling procedures, extends earlier confirmatory factor analyses of the Symptom Checklist-90--Revised Depression and Additional Symptoms scales conducted by Newmann, Engel, & Jensen (1990, 1991a, 1991b). Findings show that each measurement approach leads to different conclusions about the age-depression relation, with only the latent variable measurement showing a significant increase in depressive symptoms with advancing age. Implications of these findings for future research on aging depression are discussed. PMID- 8726378 TI - Age and sex differences in strategies of coping and defense across the life span. AB - Age and sex differences in the use of coping and defense strategies were examined in life-span sample of 381 individuals. Participants responded to 2 self-report measures assessing mechanisms of coping and defense and measures assessing their level of cognitive complexity. Older adults used a combination of coping and defense strategies indicative of greater impulse control and the tendency to positively appraise conflict situations. Adolescents and younger adults used strategies that were outwardly aggressive and psychologically undifferentiated, indicating lower levels of impulse control and self-awareness. Women used more internalizing defenses than men and used coping strategies that flexibly integrated intra-and interpersonal aspects of conflict situations. Taken together, findings provide evidence for the age- and sex-specific use of strategies of coping and defense, suggesting that men and women may face different developmental tasks in the process toward maturity in adulthood. PMID- 8726379 TI - Age differences in retrieval: further support for the resource-reduction hypothesis. AB - Age differences in processing resources seem salient to age-related declines in secondary (or "recent") memory. Community-dwelling adults (N = 90, ages 30-80) completed 4 memory tests: Wechsler Memory Scale-Revised (WMS-R) Logical Memory (LM), Cowboy Story (CS), WMS-R Visual Reproductions (VR), and Extended Complex Figure Test (ECFT; Fastenau, in press). Two space-capacity measures (WMS-R Digit Span and Visual Memory Span) and 4 processing speed measures (cancellation and mental-tracking tasks) assessed processing resources. A statistical control procedure was used to isolate retrieval efficiency and measures contributions of age and processing resources to retrieval. A negative relationship between age and retrieval efficiency emerged on all measures (p < .05). The age effect was reduced 60% on LM and CS when processing resources were controlled, eliminated for VR, and unchanged on ECFT. It is possible that visual-spatial retrieval requires fewer processing resources than does verbal retrieval. PMID- 8726380 TI - Perceptions of baby talk, frequency of receiving baby talk, and self-esteem among community and nursing home residents. AB - Community-living seniors (n = 113) and nursing home residents (n = 43) provided their impressions of baby-talk and neutral-talk scenarios and completed measures of functional health, need for succorance, and self-esteem. Two orthogonal dimensions were found in perceptions of baby talk: Warmth and Superiority. The personality trait of need for succorance was consistently associated with perceptions of warmth in baby talk, whereas functional health, age, and institutionalization were associated with perceptions of superiority. Significant interactions were found between perceptions of baby talk and frequency of receiving baby talk in the prediction of self-esteem, providing suggestive evidence for previously expressed concerns about potentially harmful effects of receiving baby talk on self-esteem among seniors who have negative perceptions of baby talk. However, older persons with positive perceptions of baby talk reported higher self-esteem when they frequently received baby talk, in accordance with person-environment theory. The self-esteem interaction for men occurred on the Superiority dimension, whereas the interaction for women occurred on the Warmth dimension. PMID- 8726381 TI - Weight changes in caregivers of Alzheimer's care recipients: psychobehavioral predictors. AB - Relationships of changes in body mass index (BMI) were examined with changes in psychobehavioral variables in spouse caregivers of individuals with Alzheimer's disease (n = 81) and matched spouses of controls (n = 86). Men caregivers had significantly greater BMI and obesity than men controls at both times. Over 15-18 months, women caregivers gained significantly more weight than did women controls. A trend for greater obesity occurred in women caregivers than in women controls at follow-up. Although weight gain was not related to psychobehavioral variables in controls, in men caregivers decreased perceived control and increased fat intake explained significant variance in weight gain. In women caregivers, increased anger control and increased calories explained weight gain. Such caregivers may be at risk for health problems. PMID- 8726382 TI - On the mechanisms of plasticity in young and older adults after instruction in the method of loci: evidence for an amplification model. AB - A meta-analytic review of the literature points out that young adults benefit more from instruction in mnemonic techniques than do older adults. In a study on memory plasticity after instruction in the method of loci, it was found that the cognitive mechanisms of plasticity in young and older adults are largely identical, with the age-related variables of speed of mental operations, associative memory, and number of list rehearsals as the core influences on plasticity. The data fit an amplification model of plasticity, in which variables positively associated with pretest performance and negatively associated with age are positively related to plasticity. Also, older adults were found to comply less with instructions, and when complying, to apply the method of loci correctly less often. Noncompliance, incorrect strategy use, and amplification might explain the adult age differences in treatment gain after instruction in a mnemonic technique. PMID- 8726383 TI - Acquisition of prototype-based information about social groups in adulthood. AB - Two experiments examined adult age differences in the ability to acquire prototype-based information about a fictitious social group. Young and older adults were presented with 60 descriptions of people who varied in their similarity to a prototypical group member along 12 (Experiment 1) or 10 (Experiment 2) feature dimensions. The prototype represented either an arbitrary combination of features or a coherent set of features based on evaluative content. Younger adults generally performed better than the older adults in learning the arbitrary prototype, whereas age differences were absent or in favor of the older adults when the prototype consisted of evaluatively consistent features. The authors argue that the results can be explained by the age-related variations in the reliance on automatic processing mechanisms and the congruence of these processes with the demands of the task across prototype conditions. PMID- 8726384 TI - Demonstration of vitamin D receptor transcripts in actively resorbing osteoclasts in bone sections. AB - The effects of the active metabolite of vitamin D, 1,25 dihydroxyvitamin D3 (1,25D), are mediated via the vitamin D receptor (VDR). 1,25D is known to have profound effects on bone resorption, but proof that the human osteoclast expresses VDR in vivo is absent. Receptors have been demonstrated in osteoblasts, and it has been generally accepted that the effects of 1,25D on formed osteoclasts are mediated via osteoblasts. Using conventional riboprobe in situ hybridization, VDR transcripts were readily detectable in osteoblasts within sections taken from normal bone and several actively remodelling bone tissues, namely, Paget's disease, renal hyperparathyroidism, and healing fracture callus. However, VDR transcripts also appeared to be present at low levels within osteoclasts from two pagetic samples and two hyperparathyroid samples. To examine this latter finding further, we have used the novel technique of in situ-reverse transcriptase-polymerase chain reaction (IS-RT-PCR) for specific amplification and detection of VDR mRNA within sections taken from the same conditions described above, and also from osteoclastoma samples. As expected, VDR transcripts were amplified and detected in osteoblasts and marrow cells, but were also prominently found in osteoclasts at approximately 50% of the level detected in osteoblasts in normal bone and at 60% in the active bone tissues. This suggests that in addition to effects on osteoclast precursors and those mediated via osteoblasts, 1,25D could exert direct effects on the active bone resorbing cells in vivo. PMID- 8726385 TI - Nitric oxide donor alleviates ovariectomy-induced bone loss. AB - Nitric oxide (NO) has been reported to inhibit osteoclastic bone resorption. We examined the bone-sparing effect of NO after 6 weeks of administration into estrogen-deficient rats. 30 female Sprague-Dawley rats, 12 weeks of age, underwent ovariectomy (OVX), and 5 rats were sham-operated. OVX rats were assigned to six groups (n = 5/group) treated respectively with: vehicle; 17-beta estradiol (E2); nitroglycerine (NG, NO donor); NG-nitro-L-arginine methyl ester (L-NAME, NO synthase inhibitor); combination of E2 + NG; and a combination of E2 + L-NAME. Prior to treatment and at the end of the treatment period, bone mineral density (BMD) of rats was measured by dual-energy X-ray absorptiometry (DXA) scanning. OVX animals had significantly lower BMD and femur weights in comparison to sham operated rats (p < 0.01), and this was completely prevented by the administration of E2 (p < 0.01). Administration of NG alone prevented OVX-induced bone loss (p < 0.05). The combination of E2 + NG did not further enhance the bone mass or femur weight, and the OVX-induced bone loss was not further aggravated by L-NAME. However, in the presence of L-NAME, E2 was totally ineffective in reversing the bone loss, suggesting that the protective effect of estrogens against bone loss may be mediated through NO. In summary, the results suggest that NO counteracts the bone loss associated with OVX. PMID- 8726386 TI - Adhesive properties of isolated chick osteocytes in vitro. AB - Different functions have been proposed for osteocytes over time, but it is now generally accepted that their most important task lies in the sensing of strain caused by mechanical loading on bone. The fact that mechanical strain can be sensed as deformation of the extracellular matrix or as fluid shear stress along the cell, in the space between cell membrane and extracellular matrix, requires that osteocytes have close (specialized) contact with the bone matrix. We studied to which extracellular matrix proteins isolated chicken osteocytes adhere and whether this adhesion is mediated by specific cell adhesion receptors called integrins. The adhesive properties of the osteocytes were compared with that of osteoblasts. Osteocytes (and osteoblasts) adhere to the same substrates (i.e., collagen types I and II, collagen fibers, osteopontin, osteonectin, fibronectin, fibrinogen, thrombospondin, and laminin). Cell spreading varied between substrates, from all cells rounded on thrombospondin to all cells fully spread out on osteopontin, osteonectin, vitronectin, fibronectin, fibrinogen, and laminin. The percentage of osteocytes adhered was equivalent to that of osteoblasts adhered on all substrates except osteopontin and vitronectin, where osteocytes adhered less. The adhesion of osteocytes and osteoblasts to osteopontin, osteonectin, vitronectin, and fibrinogen was strongly inhibited, and to fibronectin and laminin moderately, by an RGD peptide. No RGD inhibition was found on collagen. An antibody against chicken integrin alpha v beta 3, the monoclonal antibody (MAb) 23C6, did not interfere with the adhesion of osteocytes and osteoblasts to matrix proteins, whereas an MAb against chicken integrin subunit beta 1 (CSAT) strongly inhibited adhesion to all substrates. Labeling with osteocyte-specific MAbs (OB7.3, OB37.4, and OB37.11) also did not hinder the adhesion of osteocytes to collagen type I, vitronectin, and osteopontin. Adhesion sites on osteocytes were small compared with the large adhesion plaques of osteoblasts, as demonstrated by interference reflection microscopy and immunocytochemically by staining for vinculin. Osteocyte adhesion is analogous to osteoblast adhesion with regard to the range of extracellular matrix proteins to which they adhere. The adhesion is mediated by the integrin subunit beta 1, but other integrins or nonintegrin adhesion receptors are also involved. Osteocytes make contact with the extracellular matrix via small attachment points which colocalize with vinculin. This connection between the bone matrix and the cytoskeleton may be important for osteocytic sensing of mechanical strain, as it supplies a transduction route of extracellular (mechanical) signals into intracellular messages. PMID- 8726387 TI - Glucocorticoids decrease the production of parathyroid hormone-related protein in vitro but not in vivo in the Walker carcinosarcoma 256 rat model. AB - In 50-90% of cases, humoral hypercalcemia of malignancy (HHM) is due to tumor secretion of parathyroid hormone-related protein (PTHrP). Glucocorticoids are sometimes used as calcium lowering agents and there are in vitro results showing that glucocorticoids diminish PTHrP production. In this study we tested whether the serum-calcium-lowering effect of glucocorticoids is due to decreased PTHrP production by the tumor. As an animal and cell culture model we used the Walker carcinosarcoma (WCS) 256, a rat mammary carcinoma cell line producing PTHrP. In vitro, dexamethasone caused a dose-dependent inhibition of PTHrP production, whereby already 1-5 nmol/L revealed a significant decrease by WCS 256 cells. In contrast to these in vitro results, in WCS 256 tumor-bearing rats, dexamethasone (4 mg/kg body weight on day 4, and 1 mg/kg body weight from day 5 until day 7 after WCS transplantation; circulating dexamethasone levels > 20 nmol/L) did not decrease PTHrP production, PTHrP secretion, serum calcium, or tumor weight in vivo. We conclude that, in this PTHrP-mediated model of humoral hypercalcemia of malignancy, glucocorticoids do not decrease PTHrP production and secretion in vivo and do not show a calcium-lowering effect. PMID- 8726389 TI - Femoral neck geometry and radiographic signs of osteoporosis as predictors of hip fracture. AB - A total of 125 consecutive hip fracture patients were investigated regarding hip geometry. There were 33 men of mean age 76 +/- 10 years, and 92 women of mean age 78 +/- 9 years. Patients with previous hip surgery were excluded. Hip geometry (hip-axis length, width of collum femoris, and femoral shaft and neck-shaft angle) were registered on both plain radiographs and DEXA scans performed within 2 weeks after fracture. On the radiographs, the calcar femorale, the Singh index, and the femoral neck index (FNI) were also calculated and compared with earlier published values of bone mineral density hip in the hip fracture patients. The fracture cases were compared with controls, 192 DEXA scans and 163 radiographs, in patients without hip surgery or known hip disease. As measured on the DEXA scans we found a wider collum femoris and a wider femoral shaft in both the male and female fracture cases, compared to controls. Also, the fracture cases showed signs of osteoporosis as measured by the calcar femorale, the Singh index, and the femoral neck index. These measurements showed good correlation with bone mineral density of the hip as measured by the DEXA scans. PMID- 8726388 TI - Human osteoblasts survive and deposit new bone when human bone is implanted in SCID mouse. AB - We describe the first successful engraftment of viable human bone that maintains human osteoblast function and deposits new bone after implantation into pretreated SCID (Hu-bone-SCID) mice. The pretreatment consists of radiation before bone implantation and subsequent administration of antiasialo GM1 antisera every 7 days to deplete mouse macrophage and natural killer (NK) cell activity. The implanted bone samples were analyzed at various times up to 8 weeks. Comparison of pre- and post-implanted human specimens in untreated and pretreated SCID mice showed that while specimens underwent resorption and necrosis in untreated SCID mice, the structure of the implanted bone was preserved in the pretreated Hu-bone-SCID mice. In addition, histochemistry and histomorphometric analysis combined with tetracycline labeling showed that new bone formed in in the implants in the pretreated SCID mice. That the new bone resulted from the activity of the implanted human bone cells rather than from induction of differentiation of host mesenchymal cells into mouse osteoblasts was shown by immunohistochemical analysis with two different monoclonal antibodies (mAb), one mAb recognizing human HLA class II antigens and one recognizing human osteoblasts and osteocytes. Donor variability was observed in the implanted bone; however, bone from the same donor maintained its characteristics in all Hu-bone-SCID mice, and striking quantitative differences were seen in bone implanted from young (4-9 years) vs. adult (67-82 years) individuals. This model will be useful to investigate and compare endogenous osteoblast activity in different bone samples without the variables of isolating cells and examining them in vitro. PMID- 8726390 TI - The neuropeptide calcitonin gene-related peptide stimulates insulin-like growth factor I production by primary fetal rat osteoblasts. AB - Calcitonin gene-related peptide (CGRP)-immunoreactive sensory nerve terminals infiltrate all tissues including bone, in which CGRP may play a local regulatory role. To initiate studies on the role of this neuropeptide in bone, osteoblasts were isolated from fetal rat calvariae, treated with CGRP, and analyzed for cAMP and insulin-like growth factor I (IGF-I) production. CGRP alpha and -beta induced a cAMP accumulation in osteoblastic cells, suggesting that they express functional receptors for CGRP. CGRP induced an increase in both IGF-I transcripts and immunoreactive polypeptide. In contrast to prostaglandin E2 (PGE2) treatment, this increase was not accompanied by an augmentation in IGF binding proteins. Although PGE2 induced a more significant increase in IGF-I transcripts than did CGRP, the concentration of IGF-I polypeptide produced by osteoblasts was similar in response to both treatments. It is concluded from this study that CGRP has potent anabolic effects on osteoblasts, an observation which opens possibilities to study the potential therapeutic role of CGRP in osteoporosis. PMID- 8726391 TI - Bone density in young patients with congenital adrenal hyperplasia. AB - One of the major complications of glucocorticoid treatment is bone loss. 21 Hydroxylase deficiency is the most frequent inborn error of steroidogenesis, leading to congenital adrenal hyperplasia (CAH): synthesis of cortisol is impaired and replacement therapy is therefore mandatory. We studied the bone mineral density in a group of patients with congenital adrenal hyperplasia (CAH) on long-term glucocorticoid replacement therapy. We selected 30 Caucasian patients with CAH due to 21-hydroxylase deficiency (mean +/- SD age = 17.45 +/- 2.49 years). 22 patients had the classical CAH form and the remaining 8 had the nonclassical (late-onset) form. The mean duration of therapy was 15.20 +/- 4.04 years. Bone mineral density (BMD) was evaluated with a dual-energy X-ray absorptiometer. BMD was also measured in 73 healthy white volunteers of comparable age (17.35 +/- 2.99 years). BMD values of the spine (sBMD), total body (TBBMD), legs, and arms of CAH patients, adjusted for confounding variables (age, gender, body mass index), did not differ from those of control subjects (p = 0.86; p = 0.17; p = 0.06 and p = 0.26, respectively). sBMD and TBBMD values did not show relationships with the duration of treatment and the dose of corticosteroids. Patients with the classical form of CAH had bone density values comparable with those of patients with the nonclassical form (sBMD: p = 0.33; TBBMD: p = 0.97). Our data show that, despite long-term treatment with glucocorticoids, CAH patients have bone density values comparable with controls. PMID- 8726392 TI - Effects of expansive force on the differentiation of midpalatal suture cartilage in rats. AB - In an attempt to clarify the effects of biomechanical tensional force on chondrogenic and osteogenic differentiation of secondary cartilage, the midpalatal sutures of 4-week-old Wistar male rats were expanded by orthodontic wires which applied 20 g force for 4, 7, 10, and 14 days. The differentiation pathways in the midpalatal suture cartilage were examined by immunohistochemistry for osteocalcin, type I and type II collagen, and von Kossa histochemistry. Although the midpalatal sutures of the control animals consisted mainly of two separate secondary cartilages with mesenchyme-like cells at their midlines, type I collagen-rich fibrous tissue began to appear at day 4 and increased at the midline of the cartilage with days of experiment. At the end of the experiment, type I collagen-rich and calcified bone matrix appeared at the boundary between the precartilaginous and the cartilaginous cell layers. Most of the cartilaginous tissues were separated from each other and the midpalatal suture was replaced by osteocalcin-positive intramembranous bone and fibrous sutural tissue. These results strongly suggest that tensional force changed the phenotypic expression of collagenous components in secondary cartilage, which may reflect the differentiation pathway of osteochondro progenitor cells. PMID- 8726393 TI - Quantitation of incident spinal fractures: comparison of visual detection with quantitative morphometry. AB - The value of quantitative morphometry in detection of new spinal fractures was assessed in serial radiographs from 83 patients with osteoporosis. From vertebral landmarks on lateral spine radiographs, a computer program allowed calculation of vertebral heights and area. By comparing vertebral dimensions in the two sets of films, incident spinal fractures could be quantitated based either on the minimum criteria of 15% reduction in vertebral height (CM2) or a fall in height and area of 20% and 10% (CM1). The results of quantitative morphometry were compared with those of the consensus and individual readings of visual detection by three experienced investigators in the same paired sets of spinal films. For incident new fractures, the visual consensus method (V-C) showed a very good agreement with individual visual detection (kappa of 0.794 to 0.916) as well as with CM1 (kappa of 0.821). However, there was a poor agreement between the results of consensus reading and of detection by CM2 (kappa of 0.341), due to excessive number of fractures identified by CM2, but not by the visual method. For incident recurrent fractures, there was a poor agreement between V-C and individual visual detection, and between V-C and quantitative morphometry (kappa of 0.306 to 0.496). It was due to severe compression fractures at baseline, which caused further changes in vertebral dimensions difficult to measure accurately by either visual or quantitative morphometry. Thus, if the visual detection of fractures by a consensus of experienced investigators is considered as the "gold standard," quantitative morphometry, based on minimum reduction in vertebral height of 20% accompanied by a minimum decline in area of 10%, provides an objective detection of incident new spinal fractures but not of recurrent fractures. PMID- 8726394 TI - Functions of the M-CSF receptor on osteoclasts. AB - Macrophage colony-stimulating factor (M-CSF) receptor has been previously reported to be present in osteoclasts both at mRNA and protein levels. However, the biochemical interactions between M-CSF and its receptor on osteoclasts are less well characterized than in mononuclear phagocytes. In this study, we show that (1) 125I-labeled M-CSF ligand specifically binds to the M-CSF receptor on osteoclasts by autoradiography; (2) binding of M-CSF to the receptor stimulates protein tyrosine phosphorylation in osteoclasts by immunostaining; (3) oxygen derived free radicals produced by calvarial osteoclasts are increased by M-CSF stimulation (1.37 +/- 0.08, n = 10, P < 0.01); and (4) bone resorption in calvarial explants is enhanced by M-CSF (1.153 +/- 0.09, n = 10, p < 0.001). Thus, our data provide multiple lines of evidences that mouse calvarial osteoclasts are activated by M-CSF. These data suggest that under the conditions present in the calvarial model, M-CSF activates osteoclastic bone resorption. PMID- 8726395 TI - Immunohistochemical localization of the estrogen receptor in human osteoblastic SaOS-2 cells: association of receptor levels with alkaline phosphatase activity. AB - We have previously shown that the combination of estrogen (E2) and 1,25 dihydroxyvitamin D3 [1,25(OH)2D3] enhanced alkaline phosphatase (ALP) activity in human osteosarcoma SaOS-2 cells which had been grown in the presence of 10 nmol/L dexamethasone (SaOS + DEX cells). To determine whether this increase in ALP activity was associated with changes in receptor protein levels for E2 (ER) in individual SaOS + DEX cells, a monoclonal antibody to ER and a histochemical stain for ALP were used localize the expression of these proteins in fixed cells. Western and Northern blot analyses were used to determine whether E2 and 1,25(OH)2D3 affected immunoreactive ER protein and mRNA levels, respectively. Our results showed that immunohistochemical staining for ER was primarily nuclear, whereas histochemical staining for ALP was cytosolic. Treatment of cells with 1,25(OH)2D3, E2, or E2 + 1,25(OH)2D3 increased the levels of both ER and ALP activity, as visualized by enhanced cellular staining. Western analyses showed that 1,25(OH)2D3 and E2, separately and in combination, significantly increased ER protein levels. 1,25(OH)2D3 enhanced ER levels in a dose-dependent manner [analysis of variance (ANOVA), F = 3.91, p < 0.05]; this effect was augmented by E2 (ANOVA, F = 5.98, p < 0.005). In comparison, 17 alpha-E2 + 1,25(OH)2D3 and tamoxifen + 17 beta-E2 + 1,25(OH)2D3 did not increase ER levels compared with those obtained with 17 beta-E2 + 1,25(OH)2D3. ER mRNA levels were not significantly increased by E2, 1,25(OH)2D3, or E2 + 1,25(OH)2D3 together. In contrast, in a population of SaOS cells which had been in culture longer (approximately 40 passages more) than the previous cells, E2 + 1,25(OH)2D3 did not enhance ALP activity or ER levels above those obtained with 1,25(OH)2D3 alone. These results showed that in responsive SaOS cells, E2 enhanced both the stimulatory effects of 1,25(OH)2D3 on ALP activity and the activation of ER. Thus changes in ALP activity are associated with changes in ER levels in SaOS + DEX cells. PMID- 8726396 TI - The effect of socioeconomic status on bone density in a male Caucasian population. AB - We have examined the effect of socioeconomic status (SES) on bone density (BMD) in 201 males, aged 20-60 years. Males of lower SES (groups 4-6 vs. 1-3) from the total sample had significantly higher BMD (p < 0.05) at L2-4 and femoral neck. The difference was small but was not explained by differences in age, weight, calcium intake, family history, activity, or smoking. 45% of SES 4-6 males were involved in manual labor compared with 11% of those in SES 1-3, however, this also did not appear to account for the difference. PMID- 8726397 TI - A comparison of the skeletal effects of intermittent and continuous administration of calcitonin in ovariectomized rats. AB - The study was designed to compare the skeletal effects of intermittent and continuous administration of calcitonin (CT) in ovariectomized (OVX) rats. Female rats were sham operated or OVX at 3 months of age and treated for 6 weeks with vehicle or salmon CT. Sham-operated control rats were injected subcutaneously with vehicle on alternate days. One group of OVX rats was treated with vehicle intermittently by subcutaneous injection or continuously via Alzet osmotic minipumps. The remaining OVX rats were treated with CT by either subcutaneous injections (16 U/kg) on alternate days or by continuous infusion via minipumps at a daily dose of 8 U/kg. OVX rats treated with CT continuously were mildly hypocalcemic compared with all other groups. The proximal tibial metaphyses of vehicle-treated OVX rats were osteopenic with a cancellous bone volume at only 28% of the vehicle-treated control level. This bone loss was associated with increased indices of bone turnover such as osteoclast surface, osteoblast surface, and bone formation rate. Cancellous bone volume in OVX rats treated with CT either intermittently or continuously was significantly higher than that of vehicle-treated OVX rats, but lower than that of vehicle-treated control rats. Treatment of OVX rats with intermittent or continuous CT significantly decreased all indices of bone turnover compared with vehicle-treated OVX rats. However, osteoclast and osteoblast surfaces of OVX rats treated with CT continuously were still significantly higher than those of vehicle-treated control rats. These results indicate that intermittent and continuous administration of CT had similar skeletal effects in OVX rats. Both treatment regimens depressed bone turnover and partially prevented cancellous bone loss in the estrogen-deplete skeleton. PMID- 8726398 TI - Inositol 1-,4-,5-trisphosphate-dependent Ca2+ signaling by the recombinant human PTH/PTHrP receptor stably expressed in a human kidney cell line. AB - We previously reported the preparation and partial characterization of a series of human embryonic kidney cell lines (HEK-293) stably expressing various numbers of the recombinant human (h) parathyroid hormone (PTH)/PTH-related protein (PTHrP) receptor (Rc). Using this expression system we examined ligand (PTH or PTHrP) binding characteristics and cyclic AMP responsiveness. We have now extended these studies to investigate the calcium signal transduction pathways activated by the hPTH/PTHrP Rc. In parental HEK-293 cells, which lack endogenous PTH/PTHrP Rc, incubation with hPTH(1-34) had no effect on cytosolic free Ca2+ concentration [Ca2+]i. In HEK-293 clone C-21, stably expressing approximately 400,000 Rc/cell, PTH stimulated an increase in [Ca2+]i by Ca2+ release from intracellular stores; PTH released Ca2+ exclusively from the IP3 sensitive Ca2+ pool. Unlike previous studies, the ability of PTH to elicit both cAMP responses and [Ca2+]i transients occurred over a wide range of Rc numbers (between 400,000 and 3000 Rc/cell); both responses were always observed at PTH concentrations in the same dose range although the magnitude of the responses decrease with Rc number. Pretreatment of C-21 cells with pertussis toxin for 24 h, which significantly enhanced PTH-stimulated cAMP accumulation, did not modulate PTH stimulated [Ca2+]i transients. At each PTH concentration tested which resulted in increased cAMP levels, there was also an increase in [Ca2+]i transients. Treatment of C-21 cells with a battery of midregion and C-terminal PTH or PTHrP peptides showed no effect on either [Ca2+]i transients or cAMP accumulation, indicating a lack of functional interactions between these peptides and the form of the hPTH/PTHrP Rc stably expressed in these cells. Immunological analysis of G protein expression demonstrated the presence of Gs, Gi, and Gq in all parental and transfected cells lines examined. Taken together, these data demonstrate that the hPTH/PTHrP Rc, stably expressed in HEK-293 cells, elicits responses in both the cAMP and IP3-dependent [Ca2+]i pathways and is responsive only to N-terminal PTH/PTHrP peptides. PMID- 8726399 TI - Molecular diagnosis and follow up in myeloproliferative syndromes and acute leukemias: correlation between expression of fusion transcripts and disease progression in chronic myeloid leukemia. PMID- 8726400 TI - All-trans retinoic acid (ATRA) and the regulation of adhesion molecules in acute myeloid leukemia. AB - A review of recent information on the expression and the ATRA-driven modulation of cell surface adhesion molecules of acute myelogenous leukemia blast cells is presented. Cytofluorometric studies on fresh blast cells have demonstrated that CD11a, CD11b CD11c, CD15, CD45RO and CD54 expression is significantly lower in acute promyelocytic leukemia (APL) than is acute myeloid leukemia of other subtypes (AML). In vitro treatment with ATRA dramatically modifies the adhesion phenotype of APL blast cells, promoting a consistently striking up-regulation of CD11b, CD11c, CD15, CD65, CD54, and CD38. Which is in general, poorly demonstrable in AML. The behaviour of CD15s is variable and fully independent from CD15 and CD65 in induction experiments, suggesting a differential enzyme regulation within the selectin ligand system. ATRA is capable, in both APL and AML, of producing a switch from the high- (RA) to the low- (RO) molecular weight isoform of CD54, Moreover, treatment with this retinoid exerts a negative regulation of the membrane expression of CD49e, CD58 and CD11a in APL as well as in AML. Of particular interest is the fact that the negative effect on CD1 1a expression generates an asynchronous phenotype in APL (CD11a-, CD11b+, CD15+), undetectable on normal maturing myeloid cells. In the last part of this review the possible implications of adhesion molecule modulation in the pathogenesis of ATRA syndrome are discussed. PMID- 8726401 TI - Integrin receptors and hairy cell leukaemia. AB - The ability of a cell to recognize and specifically localise within an appropriate tissue environment is essential to the proliferation and survival of that cell. The integrin family of cell-surface adhesion-receptors are essential to such tissue localisation, allowing a migrating cell to specifically recognise, localise within-, and respond to- the cellular or extracellular matrix ligands that characterise a given tissue. We have investigated how the expression and activity of integrin receptors underlies the consistent and unusual tissue distribution of the malignant B lymphocytes of hairy cell leukaemia (HCL). In this report we review our published work in this area. Our findings are then discussed within the context of current knowledge of integrin receptors and their ligands, and in relation to the clinical features of HCL. PMID- 8726402 TI - Cytomegalovirus as a cause of pancytopenia. AB - Human cytomegalovirus, HCMV, infects most of the population by adulthood; The primary infection is often accompanied by transient neutropenia and thrombocytopenia, and is followed by a period asymtomatic viral latency. In the setting of bone marrow transplantation, however, the immunosuppressed state of the recipient enables HCMV to re-activate or to infect the individual and cause serious sequelae. These range from hepatitis and gastrointestinal disease to interstitial pneumonia and hematologic abnormalities, which are more common in the allograft. Little is currently known about the mechanisms by which HCMV causes these hematologic abnormalities. In this review, we discuss experimental models which are helping investigators understand the immunology and pathology of CMV infection. We also summarize the vivo studies of the effects of HCMV on human hematopoiesis. Several possible mechanisms that could explain the deleterious effect of HCMV on human hematopoietic function include: 1) alteration of accessory cell function by inducing the production of inhibitory cytokines; 2) perturbation of stromal cell function resulting in a decreased production of hematopoietic factors or by altering cell surface adhesion molecule expression; 3) by direct infection of the hematopoietic stem or progenitor cells. It is likely that the pathogenesis of this syndrome is multifactorial therefore requiring a broad therapeutic approach. This would include the use of the antiviral agents, hematopoietic growth factors and donor derived HCMV specific cytolytic cells. PMID- 8726403 TI - On the phosphorylation of 2-chlorodeoxyadenosine (CdA) and its correlation with clinical response in leukemia treatment. AB - The nucleoside analog 2-chlorodeoxyadenosine (CdA, Cladribine) is a chemotherapeutic agent for treatment of leukemias and lymphomas, most successfully used in hairy cell leukemia and B-cell chronic lymphocytic leukemia. CdA is phosphorylated intracellularly to its monophosphate derivative by the enzymes deoxycytidine kinase and deoxyguanosine kinase. Cell lines deficient in deoxycytidine kinase were shown to be resistant to CdA and a high deoxycytidine kinase level in combination with low 5'-nucleotidase has been proposed to partly explain the selectivity in CdA toxicity for lymphoid cells. In this report biochemical properties in CdA phosphorylation mediated by deoxycytidine kinase and deoxyguanosine kinase are reviewed and discussed in relation to the further metabolism of CdA 5'-monophosphate, the different possible mechanisms of action and the correlation with clinical response. It is concluded that much is known about the metabolism and mechanisms of action of CdA, but that the remarkable therapeutic effect in hairy cell leukemia has yet to be explicitly explained. PMID- 8726404 TI - Effects of alpha-interferon on serum beta-2-microglobulin. AB - Beta-2-microglobulin (B2M) forms the small invariable light chain subunit of class I HLA antigens on the cell membrane of all nucleated cells. During the continuous turnover of the HLA molecules, B2M is shed from the cell membrane into blood. Lymphocytes are the main source of serum free B2M. Serum B2M concentration is increased in renal diseases, various malignant diseases and some inflammatory and autoimmune disorders. In lymphatic malignancies serum B2M has significant prognostic value. Interferons (IFNs) have the ability to enhance the expression of class I and II histocompatibility antigens. Accordingly, IFNs cause a rise in formation and release of B2M. Currently, treatment with IFN alpha is used in diseases, like multiple myeloma, where serum B2M measurements are used to assess tumor burden. We have measured serum B2M levels during IFN alpha treatment in patients with both multiple myeloma and chronic myeloproliferative diseases, and IFN alpha caused a significant increase in serum B2M. It can be concluded that use of IFN alpha abolishes the value of serum B2M as an indicator of disease activity. PMID- 8726405 TI - Biological characteristics of CD7(+) acute leukemia. AB - Eighty six of 430 acute myeloblastic leukemia (AML) patients (20.0%) and forty of 173 acute lymphoblastic leukemia (ALL) patients (23.1%) had CD7 on their leukemia cells. CD7(+) AML occurred at a younger age than CD7(-) AML, and is more frequent in males. Hepatomegaly and central nervous system involvement were also more frequent in CD7(+) AML than in CD7(-) AML. The age of onset of CD7(+) ALL is also younger than that of CD7(-) ALL. Phenotypically, CD(+) AML expressed CD34, HLA DR, and TdT more frequently than CD7(-) AML while CD7(+) ALL expressed CD13/33 more often than CD7(-) ALL cells responded most significantly to interleukin 3 (IL-3), whereas most CD7(-) AML cells responded more significantly to granulocyte macrophage-colony stimulating factor (GM-CSF) and/or granulocyte (G)-CSF than to IL-3. CD7(+)sCD3(-)CD4(-)CD8(-) ALL expressed G-CSF receptor and c-kit mRNA more frequently, which is not usual in other types of ALL. P-glycoprotein (P-gp)/multi drug resistance gene (MDR1), thought to be expressed in hematopoietic stem cells, is expressed in CD7(+) AML and CD7(+)sCD3(-) CD4(-)CD8(-) ALL significantly more often than in CD7(-) acute leukemias and the CR rate and overall survival of CD7(+)AML was worse than CD7(-) AML. These data, collectively, suggest the close association of CD7(+) AML and CD7(+)sCD3(-)CD4(-)CD8(-) ALL, not only the common expression of CD7 itself but also because their phenotypical immaturity, cytokine receptor expression, P-gp/MDR1 expression and clinical manifestations including the frequent occurrence in males and the poor prognosis. We propose that CD7(+) acute leukemia is an hematopoietic stem cell leukemia which may be separate entity. PMID- 8726406 TI - Intensive therapy and autologous stem cell transplantation for Hodgkin's disease in first relapse after combination chemotherapy. AB - Data from a number of transplant centers has shown that several intensive therapy regimens, supported by autologous stem cell transplantation, have the capability to produce durable responses in a proportion of patients with Hodgkin's disease progressive after combination chemotherapy. Although many questions regarding the optimal use of autotransplantation remain unanswered, the issue of the preferred timing at which to apply transplantation is of critical importance in planning therapeutic strategies for patients with this disease. This paper will focus on the timing options for autotransplantation in Hodgkin's disease. In the absence of a formal Phase III study comparing conventional salvage therapy versus autotransplantation in first relapse patients, the encouraging results from our center and others support the use of transplantation at the time of first relapse after combination chemotherapy. Non-relapse mortality is low in this setting, and the primary problem has been recurrent disease despite transplantation. Risk factors for both disease recurrence, as well as for the probability of progression-free survival, can be defined based on biologic features present at the time of first relapse after chemotherapy, and may provide a basis for improving the current transplant results for first relapse patients. Prolonged follow-up will be important to define the incidence and risk of late toxicities in autografted patients. PMID- 8726407 TI - Patterns of ganglioside expression in B cell neoplasms. AB - Twenty seven B cell neoplasms were examined by high performance thin layer chromatography (HPTLC) and immune thin layer chromatography (ITLC) to determine ganglioside expression. Patterns of expression in the cells were compared with conventional morphology, genotype, and glycoprotein immunophenotype. Patterns of ganglioside expression were found for each of the tumor types analyzed (5 acute lymphoblastic lymphomas (ALL), 5 Burkitt's Lymphomas (BL), 4 chronic lymphocytic leukemias (CLL), and 3 diffuse poorly differentiated lymphomas (DPDL), 7 diffuse histiocytic lymphomas (DHL), and 3 multiple myelomas (MM). GM3 was the predominant ganglioside found in all B cell neoplasms except multiple myeloma where GM2 was equivalent to GM3. GM1 was detected by ITLC in all B cell tumors, but significant amounts were found by HPTLC only in ALL, CLL, and DHL. Small amounts of GD3 and GD2 were found in several B cell neoplasms. Significant amounts of other gangliosides were not found. The expression of GM2 on the MM cell lines, a cell type derived from outside of the nervous system, is unusual. This high level of expression was also seen in metabolic labeling studies. GM2 was readily detectable in the SKMM1 human multiple myeloma cell line by flow cytometry and served as a target for human complement-mediated cytotoxicity. Although the functions of gangliosides are largely known, the patterns of gangliosides found for this system of human B cell malignancies may serve to provide targets for specific immunotherapy and clues to their functions. PMID- 8726408 TI - Heterogeneity of B lineage acute lymphoblastic leukemias (B-ALL) with regard to their in vitro spontaneous proliferation, growth factor response and BCL-2 expression. AB - The spontaneous proliferation and the effects of 8 various growth factors (GF) were evaluated on leukemic cells from 27 patients with B-lineage ALL. Two groups of ALLs were distinguished. ALLs from group I (21 patients) exhibited a low spontaneous proliferative rate and were stimulated by IL-3 + IL-7 +/- SCF and/or LIF, while ALLs from group II (6 patients) had a high spontaneous proliferative rate and did no longer require this combination of GFs for proliferation. No effect of bFGF, IGF-I, IL-10 and IL-11 alone or in combination, was observed. Such differences in the behaviour of B-ALLs indicated that the GF requirement of ALL blasts was not related to the presence of serum in the culture nor to the pattern of reactivity of ALL blasts for B lymphoid markers or CD34 antigen. Furthermore, we showed in 1/9 cases that high proliferation might be due to an overexpression of the bcl-2 proto-oncogene and to the acquisition of an autocrine secretion. PMID- 8726409 TI - Expression and production of cytokines by heterohybrids and their parental B cells in CLL. AB - Three hybrids derived from CD5+ B cell chronic lymphocytic leukemia (B-CLL) and their parental B cells were studied for phenotypic evolution, immunoglobulin (Ig), tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) secretion. When phenotypic evolution was examined, hybrids showed the loss of classical B cell markers, indicating that they follow the same pattern of phenotypic differentiation as normal B cells. Hybrids displayed spontaneous high Ig secretion, which did not appear to be modified through stimulation by phorbol 12 myristate 13-acetate (PMA), recombinant interferon-gamma (rIFN-gamma) and Staphylococcus aureus Cowan I (SAC). Parental cells secreted minimal amounts of Ig spontaneously or through IFN-gamma and SAC stimulation, whereas PMA succeeded in increasing this secretion. An opposite pattern was observed when TNF-alpha and IL-6 secretion an expression at the mRNA level were assessed in hybrids and parental cells. TNF-alpha and IL-6 were spontaneously secreted by parental cells and this secretion was increased after PMA and SAC stimulation, both cytokine secretion and expression at the mRNA level were negative in hybrid cells. The absence of expression of these cytokines could be explained either by chromosomal loss or by down regulation. These results indicate that when parental CLL cells are induced to differentiate in the heterohybrid model, they acquire high spontaneous secretion of Ig, lose the classical B cell phenotypic markers and down regulate the expression of the cytokines studied. PMID- 8726410 TI - Mechanisms of inhibition of IL-6-mediated immunoglobulin secretion by dexamethasone and suramin in human lymphoid and myeloma cell lines. AB - The cytokine IL-6 has been proposed as an autocrine growth factor in multiple myeloma, and is also required for stimulation of immunoglobulin production and secretion in normal plasma cells and myeloma cells. In this study, we showed that secreted IL-6 is detectable by Western blot analysis in a panel of lymphoid and myeloma cell lines. Previous studies in our laboratory have shown that dexamethasone and suramin inhibit cell proliferation and IL-6-mediated immunoglobulin secretion in various lymphoblastoid and myeloma cell lines. In the present study, we present study, we present data to examine mechanisms by which dexamethasone and suramin inhibit IL-6-mediated immunoglobulin secretion in the lymphoid cell line SKW 6.4. Cells treated with rIL-6 or the IC10 concentration of dexamethasone respectively undergo a doubling of intracellular IgM. Moreover, rIL 6 and dexamethasone additively stimulate cells to accumulate intracellular IgM. In contrast, cells treated with the IC10 concentration of suramin undergo no significant alteration of total cellular IgM, and do not respond to IL-6 with an increase in intracellular IgM. Northern blot analysis demonstrates that cells treated with exogenous rIL-6 and/or dexamethasone respectively undergo a coordinate one to three fold increase of kappa and mu chain mRNA expression, while there is a 30-40% decrease of kappa and mu chain mRNA when cells are treated with suramin and suramin plus rIL-6. Western blot analysis shows that levels of intracellular IL-6 modestly increase when cells are treated with exogenous rIL-6, whereas treatment with dexamethasone plus rIL-6 causes a 70% decrease of immunoreactive IL-6 protein in comparison with untreated cells. An rtPCR analysis of IL-6 mRNA expression shows an abolished signal in response to dexamethasone or rIL-6 and/or dexamethasone. Using a flow cytometric assay, it is demonstrated that suramin inhibits IL-6 binding to its receptor. Taken together, these results indicate that SKW 6.4 cells treated with rIL-6 and/or dexamethasone undergo increased expression of IgM mRNA leading to increased intracellular IgM levels. Treatment with suramin or suramin plus rIL-6 does not alter the IL-6 protein level or the mRNA levels for IL-6 and IL-6 receptor. Suramin treatment causes a moderate decrease in IgM mRNA, and this is associated with a decreased intracellular level of IgM in SKW 6.4 cells. Overall these findings support the concept that IL-6 is an autocrine factor for immunoglobulin production and secretion in myeloma cells. Suramin interferes with IL-6 binding to its receptor and/or decreases IL-6 receptor expression. Dexamethasone has neither of these effects on IL-6 receptor expression or IL-6 binding to its receptor, and we postulate that it acts through a block in secretion or in degradation of intracellular immunoglobulin by decreasing IL-6 mRNA expression and IL-6 protein content. These studies suggest that the combination of suramin and dexamethasone not only synergistically growth inhibit myeloma cells but also act in concert to inhibit immunoglobulin secretion and represent a therapeutic approach worthy of further investigation. PMID- 8726411 TI - Macrophage expression of interleukin-1 in lymphoepithelioid cell ("Lennert's") lymphoma. AB - Macrophage activation was studied in three cases of a genuine form of T cell non Hodgkin's lymphoma particularly rich in epithelioid histiocytes, the so-called "lymphoepithelioid cell lymphoma" or "Lennert's lymphoma". Host tumor infiltrating macrophages actively produced Interleukin-1 as demonstrated by immunohistochemistry. The activated histiocytes also contained intracytoplasmic tumor cells which were either intact or at various stages of apoptosis. We postulate that in Lennert's lymphoma, tumor cells are capable of activating host macrophages. Initial macrophage activation is followed by IL-1 production with recruitment of additional macrophages accounting for the characteristic histological appearance of this tumor. The activated macrophages are also engaged in a phagocytic antitumoral response. Future studies should investigate if this host response can be potentiated. PMID- 8726412 TI - Primary Mediastinal B-cell lymphoma with sclerosis: clinical and therapeutic evaluation of 22 patients. AB - In the last decade, there have been several reports on what is now recognized as a new clinical and pathological entity termed primarily mediastinal B-cell lymphoma (PMBCL) with sclerosis. This lymphoma presents unique clinical characteristics with an aggressive outcome and, at present, the best approach seems to be a combination of chemotherapy and radiotherapy. Between June 1989 and September 1994, twenty-two previously untreated patients with PMBCL with sclerosis were treated with a combination of third-generation chemotherapy regimen (MACOP-B or F-MACHOP) and mediastinal irradiation. All the patients presented with bulky mediastinal involvement; the radiologic clinical stage with evaluation of tumor size included computed tomography and Gallium-67-citrate SPECT. Twenty-one patients (95%) achieved a complete response and only one was resistant to treatment. Regarding 67Ga SPECT, 6 patients, including the nonresponder, showed persistent abnormal 67Ga uptake after chemotherapy; however after the mediastinal radiotherapy, all the patients except for the nonresponder were 67Ga-negative. The overall survival was 87%, with a median follow-up of 24 months from the time of diagnosis. Two of the patients who achieved complete response relapsed 7 and 10 months after completion of treatment, respectively. The relapse-free survival rate was 89% at 62 months (median 20 months). In patients presenting with bulky mediastinal PMBCL with sclerosis combined modality treatment using third-generation chemotherapy regimens and radiotherapy induces a good remission rate with greater than 80% chance of surviving disease-free, at 2 years. A longer follow-up before definitive conclusions are drawn is still warranted. PMID- 8726413 TI - Salvage therapy with fludarabine in patients with progressive B-chronic lymphocytic leukemia. AB - Thirty patients with B-chronic lymphocytic leukemia, aged 45-82 years, were treated with fludarabine. CLL was diagnosed 8-120 months earlier. The patients had been exposed to a median of 3 different regimens before treatment with fludarabine, and all had progressive disease when they entered the study. Among the 30 patients, 1 had a metastatic carcinoma and 7 patients with WHO performance status 3 died before the second cycle of fludarabine treatment could be given. The remaining 22 patients were considered eligible for response evaluation. The response rate was 32% with 1 complete response and 6 partial responses. However, seven patients achieved stable disease and 8 progressed. The median survival for responders was 24 months and for non-responders 9 months. Response to treatment was correlated with low tumor burden and performance status. In a total of 94 treatment courses, 17 febrile episodes were registered in 10 patients. We conclude that treatment with fludarabine can be useful in patients with progressive and refractory disease. PMID- 8726414 TI - Fludarabine monophosphate in refractory B-chronic lymphocytic leukemia: maintenance may be significant to sustain response. AB - In the present study we report our results on the efficacy of Fludarabine monophosphate in 20 B-chronic lymphocytic leukemia (CLL) patients, refractory to conventional chemotherapy. Of the 20 patients 14 were males and 6 females with a median age of 58 years (44-70). Eight had Binet stage B and 12 stage C. They were previously treated with chlorambucil, prednisone, mini-CHOP or irradiation. Their disease duration prior to fludarabine administration was 49 months (7-180). Fludarabine was given at a dose of 25 mg/m2 daily, for five consecutive days, monthly for six months and if responding for six additional months. Treatment was administered on an outpatient basis. Complete response (CR) was observed in 7 patients (33%) and partial remission (PR) in 5 (25%). Of the complete responders 5 were males and 2 females with a median age of 60 years (range 55-68); three of them had stage B and 4 stage C disease; the median number of fludarabine courses for achieving CR was 3 (range 2-5). In all CR patients a residual monoclonal CD5/CD19 positive lymphocyte population was found in the peripheral blood. All CRs relapsed shortly after discontinuation of therapy within 12 months. The main toxicity observed was upper respiratory tract and/or pulmonary infections in 8 patients, requiring hospitalization. Among the CRs one patient died during the administration of the third course of therapy, due to a severe hypersensitivity reaction with Stevens-Johnson syndrome. The importance of maintenance therapy is also stressed as PR was sustained in some patients using 3 day cycles every 2-4 months. One patient was maintained in this fashion for 60 + months. This study showed that fludarabine is effective in CLL patients refractory to conventional chemotherapy thus it may be given as the treatment of choice if such patients still require treatment. PMID- 8726415 TI - Clinical and immunological studies in advanced cancer patients sequentially treated with anti CD3 monoclonal antibody (OKT3) and interleukin-2. AB - CD3 engagement has been used as a surrogate for antigen-specific stimulation to trigger T cell effector functions. Exogenous IL-2 has been used to prolong and amplify CD3-induced T cell activation. Previous studies have been shown that CD3 reactivity is increased in cancer patients with preactivated (> 10% HLA-DR+) T cells in the peripheral blood. In this study, we report 9 courses of a single infusion of anti-CD3 mAb (OKT3) followed by continuous infusion of intermediate dose IL-2 in 4 cancer patients [2 multiple myeloma (MM), 1 B-cell lymphoma (NHL), 1 metastatic melanoma (ME)] with advanced disease and > 10% HLA-DR+ T cells in the peripheral blood. An increase of lymphocytes, equally distributed between CD4+ and CD8+ subsets, was observed during treatment. Activation was phenotypically documented by the emergence of CD25+ cells in the peripheral blood. Unexpectedly, functional studies [including proliferation to mitogens (PHA, OKT3) and cytotoxicity assays (NK and LAK activities)] did not parallel phenotypic data and a slight decrease of all functions was observed after OKT3 and IL-2 treatment. OKT3 and IL-2 infusions were well tolerated and no limiting toxicity was observed. The treatment did not revert tumor progression in the 2 patients with progressive disease (NHL, ME) and had only minimal effects in the 2 MM patients with stable disease. These data indicate that the sequential administration of OKT3 and IL-2 had no anti-tumor activity in this small series of patients with advanced cancer who were selected for treatment because of an increased number of HLA-DR+ T cells in the peripheral blood. A discrepancy was observed between the emergence of CD25+ T cells and the clinical outcome. PMID- 8726417 TI - Simultaneous occurrence of large B-cell non-Hodgkin lymphoma and myelodysplastic syndrome rapidly evolving into acute myeloblastic leukemia. AB - We describe a case of simultaneous occurrence of large B-cell non-Hodgkin lymphoma and myelodysplastic syndrome in the absence of previous chemotherapy or radiotherapy. After initiation of steroid treatment, the myeloid clone showed a rapid increase in both the bone marrow and peripheral blood with transformation into acute myeloid leukemia. The diagnosis were confirmed by immunophenotypic studies performed in the histologic sections of the lymph node, as well as in bone marrow and peripheral blast cells. This case may be indicative of potential down-regulation of a malignant myeloid clone induced by the malignant lymphoid clone. PMID- 8726416 TI - Antitumor effect in cyclosporine A- or FK506-treated mice after syngeneic bone marrow transplantation. AB - We investigated the possibility for induction of graft-versus-tumor (GVT) effects in cyclosporine A (CsA)- or FK506 (FK)-treated DBA/2 mice after syngeneic bone marrow transplantation (BMT). For in vitro assays of spleen cells, the CsA treated mice had more enhanced cytotoxic activity against YAC1 and P388, while the FK-treated animals had more against P815, YAC1, and P388. IL-4 mRNA expression was detected in spleen cells of the FK-treated mice and IL-6 mRNA expression was clearly detected in both the treated groups. Concerning GVT effects, FK had more pronounced immunostimulatory potential than CsA in this experimental setting using DBA/2 mice. In tumor-loading in vivo experiments, we could not show any antitumor effect on survival. However, this immunostimulation could be expected to eradicate the minimal residual disease after autologous BMT and autologous peripheral blood stem cell transplantation. PMID- 8726419 TI - Cutaneous manifestation of leishmaniasis 40 years after exposure in a patient with chronic lymphocytic leukaemia. AB - Most individuals infected with the protozoan parasite Leishmania do not demonstrate overt disease, because of effective immune protection produced by T cell mediated immunity. Acquired defects in T-cell responses may lead to emergence of leishmaniasis many years after exposure in endemic areas. We describe a case of a 75 year old man, who presented with a cutaneous manifestation of leishmaniasis 40 years after exposure, co-incident with the diagnosis of chronic lymphocytic leukaemia. PMID- 8726418 TI - Fulminant 2-chlorodeoxyadenosine-related peripheral neuropathy in a patient with paraneoplastic neurological syndrome associated with lymphoma. AB - 2-chlorodeoxyadenosine (2-CdA) has been demonstrated to be a neurotoxic agent when used at significantly greater doses than currently recommended for clinical use. In this report we describe a case of a 37-years-old man lymphoplasmacytoid malignant lymphoma and pre-existing paraneoplastic neurological syndrome who died of an apparent rapidly progressive sensorimotor peripheral neuropathy after completing treatment with two courses of low-doses of 2-CdA. PMID- 8726420 TI - Teaching cases from the Royal Marsden and St Mary's Hospital case 11 dysplastic neutrophils in an African women. PMID- 8726421 TI - Surgical anatomy of the nose. AB - Knowing the details of nasal anatomy is absolutely necessary when undertaking the responsibility of rhinoplasty surgery. It is the essential first step. Careful study of these details makes for a more confident, prepared practitioner. PMID- 8726422 TI - Dynamic interplays during rhinoplasty. AB - One of the complicating factors in rhinoplasty procedures is its intricate dynamics. Not only is there a complex interaction between the soft tissue and nasal bone frame on each nasal zone, there is an intriguing interaction between the different zones of the nose and the surrounding facial structures. Understanding these interplays is mandatory to proper design and execution of the aesthetic surgical plans. This article has covered the dynamic effects that occur along the different nasal zones, elucidated the control that the nasal frame has on the soft tissue, and reviewed the different ways in which the intended aesthetic objectives can be obtained. PMID- 8726423 TI - An aesthetic dorsum. The CATS graft. Cartilaginous autogenous thin septal. AB - Reconstruction of the dorsum following a Joseph hump removal can be accomplished with a properly thinned septal graft. This will camouflage the residual twists in the dorsal septum, the irregularities at the osseocartilaginous function, and an open roof. It is particularly useful in those patients with a prominent hump but can be used in those patients with narrow noses in whom one does not wish to do an infracture following the hump resection. PMID- 8726424 TI - Rethinking the logic and techniques of primary tip rhinoplasty. A perspective of the evolution of surgery of the nasal tip. AB - Historically, destructive tip-shaping and positioning techniques, although effective to some degree, have created large numbers of secondary deformities. Tip grafts, necessary in secondary rhinoplasty, have been applied widely in primary rhinoplasty, introducing additional variables and a significant reoperation rate. Nondestructive tip-shaping and positioning techniques offer the surgeon a wider range of alternatives, more control and predictability, and a lower reoperation rate. We are making major changes in the way we think about primary tip rhinoplasty. Most importantly, we are learning (or relearning) that preservation of normal anatomy and structural integrity is a surgical principle epitomized in primary rhinoplasty, and that when we can achieve the desired result using that normal anatomy and preserving its structural integrity, the long-term result is more predictable. PMID- 8726425 TI - Use of spreader grafts in the external approach to rhinoplasty. AB - The challenge for the modern rhinoplasty surgeon is to create a nose that appears natural and balances with the individual's face and ethnicity. The use of spreader grafts in endonasal rhinoplasty as described by Sheen has been expanded by the external approach to (1) straighten and buttress weakened L-struts in a high dorsally deviated septum, (2) recreate dorsal aesthetic lines, (3) reconstruct a narrow middle vault, and (4) restore the internal nasal valve. The advantages of the external approach in the use of septal spreader grafts includes ease of harvest, accurate contouring, and precise placement and suture stabilization to minimize postoperative graft displacement. The versatility of this technique allows the surgeon to attain consistent functional and aesthetic results in the management of dorsal mid-vault problems in primary and secondary rhinoplasty. PMID- 8726426 TI - Alar base abnormalities. Classification and correction. AB - Abnormalities of the alar base may be a consequence of base deformities, a secondary result of reduction of nasal base projection, or a consequence of maxillary abnormalities. A thorough clinical observation coupled with information procured from the analyzed lifesized photographs provides a consistent method for the evaluation of the alar base and the structures that affect its position. A critical step in choosing the surgical technique for correction is the classification of abnormalities. A systemic approach to such a classification is presented in this article and is accompanied by a description of the dynamic changes and optimal surgical procedures that would best benefit patients within each category. PMID- 8726427 TI - Surgery of the nasal septum. AB - When nasoseptal dislocation or deviation contributes to nasal obstruction or external deformity, successful nasal surgery should include septal restructuring, maintenance or restoration of adequate nasal support, and stabilization of the reconstructed elements. PMID- 8726429 TI - The short nose. AB - Lengthening the short nose is arguably the most difficult operation in aesthetic rhinoplasty. Open and closed operative techniques can achieve absolute lengthening. In the open technique, maximum lengthening is achieved by releasing the mucoperichondrium, which has to be elevated bilaterally from the septum; releasing the soft tissue between the upper and lower lateral cartilages; and attaching a batten graft to the septum to project the tip cartilages caudally. The closed technique uses a hemitransfixion incision and elevation of the mucoperichondrium bilaterally, followed by attachment of a batten graft. The apparently short nose also can be treated by derotating the tip, correcting the columellar labial angle, and altering the location of the nasion as well as the nasofacial angle. PMID- 8726428 TI - Rhinoplasty with advancing age. Characteristics and management. AB - The aging nose presents several unique challenges to diagnosis and management. Although classically it presents as a drooping nasal tip secondary to loss of tip support with a relative dorsal hump, there are associated changes in the nasal skin, bony architecture, and airway that mandate consideration. Methods of correction emphasize restoration of tip support with nondestructive suture techniques. Use of osteotomies is minimized. Airway patency is improved or maintained by restoring the internal nasal valve, often with dorsal spreader grafts. Finally, special consideration is given to the patients' unique expectations and motivation. Rhinoplasty in the patient with advancing age is a rewarding operative procedure if one follows the guidelines delineated here to optimize the functional and aesthetic results. PMID- 8726430 TI - Alloplastic nasal and perialar augmentation. AB - Our favorable experience with use of porous hydroxyapatite granules to augment the craniofacial skeleton (more than 200 patients during an 8-year period) has led us to use this method to augment the nasal skeleton in selected cases. Extensive experience has been achieved in augmenting the perialar, maxilla, and glabellar area with very favorable results. A much more limited experience has been gained in augmenting the nasal dorsum, and this method must be considered investigatory at present. PMID- 8726432 TI - Surgery of the non-Indo-European face. AB - The severely deviated nose represents a complex cosmetic and functional problem. This article analyzes the types of deformities present in a severely deviated nose, reviews the existing methods of correction, and proposes a new method of internal cartilage splinting to correct the more severe deformities. PMID- 8726431 TI - The severely twisted nose. Treatment by separation of its components and internal cartilage splinting. AB - Adequate treatment of the deviated nose requires a detailed evaluation of the deformity present. A new method of component separation with internal cartilage splinting has been used with success to correct the severely deviated nose. The separation of the components allows detection and correction of all areas of cartilage deviation. The internal cartilage splinting allows long-term support and avoidance of mid-vault collapse. PMID- 8726433 TI - Clinical frontiers of interactive image-guided neurosurgery. Introduction. PMID- 8726434 TI - Image-guided spine surgery. AB - Conventional imaging techniques are of limited usefulness in spinal navigation, due to their lack of a "three-dimensional perspective." An interactive image guided approach has been developed that provides a three-dimensional image-space representation of surgical space, using a specially designed referencing system and computer workstation. The system is described and results from laboratory tests and clinical cases are reported. It is concluded that interactive image guided stereotaxis can be a valuable tool in spinal localization because of its degree of precision and the increased margin of safety it can offer. PMID- 8726435 TI - Image-guided surgical techniques for infections and trauma of the central nervous system. AB - The value of image-guided stereotactic systems is directly dependent on the ease and speed of their use. In the past, most stereotactic techniques were complicated and timely to set up; thus, they were used exclusively for either resecting neoplasms or for neurologic function. However, current systems equipped with advanced registration techniques are much simpler and faster to employ, and indications for their use are rapidly increasing. We describe an advanced image guided navigation system and provide examples of its successful use in neurosurgical treatment of central nervous system infection and trauma. PMID- 8726436 TI - Vascular malformations of the brain. Surgical management using interactive image guidance. AB - This article describes the use of interactive image guidance in the surgical management of vascular malformations (VMs). Indications include VMs located in or around eloquent brain regions such as speech and motor sensory areas. The image guidance provides a three-dimensional view of the actual vascular malformations through computer simulation and an accurate means of determining its position in relation to adjacent brain structures (anatomic localization). PMID- 8726437 TI - The evaluation and image-guided surgical treatment of the patient with a medically intractable seizure disorder. AB - Over the past decade, there has been a remarkable increase in awareness of the role surgery may play in the management of the patient with medically intractable epilepsy. This article explores the role of emerging interactive image-guided techniques in noninvasive and invasive preoperative seizure evaluation and the planning and execution of surgical treatments. PMID- 8726438 TI - Image-guided surgery of epilepsy. AB - Interactive image-guided techniques used in conjunction with three-dimensional images allow accurate planning and performance of a variety of neurosurgical procedures. The authors have used the frameless stereotactic Allegro Viewing Wand System to provide real-time correlation of the operating field and computerized images in over 200 neurosurgical operations carried out for intractable epilepsy. The authors experience shows that the viewing wand system is most helpful as an adjunctive navigational device in the microsurgical treatment of epilepsy. PMID- 8726439 TI - A technique for interactive image-guided neurosurgical intervention in primary brain tumors. AB - Interactive image-guided neurosurgical techniques allow safer and more complete cytoreduction of gliomas. This is most significant for low-grade tumors, whose configurations and margins are perhaps better appreciated by reference to registered MR images rather than by reliance on direct visualization using microscopic illumination. Spatially registered electro-physiologic recordings of intraoperative cortical stimulation to map language and motor function can increase the margin of safety for performing radical resections. By individualizing approaches and optimizing results, these technologies promise a new degree of standardization of outcome after resective surgery for all glial tumors. PMID- 8726440 TI - Intraoperative navigation during resection of brain metastases. AB - This article reviews the authors' experience with image-guided surgery for brain metastases and discusses specifically the impact of the frameless viewing wand system on standard craniotomy techniques for this disorder. Topics discussed include patient selection, interactive image-guided neurosurgical resection of brain metastases, and other image-guided neurosurgical systems. PMID- 8726441 TI - Surgical management of convexity and falcine meningiomas using interactive image guided surgery systems. AB - Interactive image-guided surgery systems can provide useful navigation information to assist craniotomy for meningiomas of the convexity and falx. When using MR imaging for navigation, interactive image-guided surgery can provide visualization of the relationship of tumor to venous anatomy, which may reduce the risk of debilitating infarction. Cortical surface anatomy can also be assessed, helping to estimate risk to functionally expressive areas. PMID- 8726442 TI - Image-guided surgery for skull base neoplasms using the ISG viewing wand. Anatomic and technical considerations. AB - Image-guided surgery of the cranial base requires a knowledge of both surgical anatomy and the technical aspects of the particular system being used. The authors report on their experience using the ISG Viewing Wand over the last two years, concentrating on the technical considerations for the use of the system and lessons learned. Image-guided surgical systems may well replace the current forms of intraoperative imaging used for anatomic orientation and localization and are considered a valuable addition to the armamentarium of the skull base surgeon. PMID- 8726443 TI - Skull base tumors. A critical appraisal and clinical series employing image guidance. AB - Two of the most significant developments in neurosurgery over the past ten years have been the application of intraoperative navigational units and the widespread use of "skull base" surgical techniques. These developments have independently revolutionized the way in which neurosurgeons approach intracranial pathology. The combination of these two developments has had a profound effect in our clinical practice to improve outcome and extend our capabilities. This ability to see around the corner, "x-ray" vision as some have called it, provides an added measure of safety for the patient while allowing the surgeon to treat the lesion in a more aggressive fashion. PMID- 8726444 TI - Functional magnetic resonance imaging for intracranial navigation. AB - Functional MR imaging can provide accurate anatomic and physiologic localization of human cortical function. This new method of noninvasive cortical mapping appears to be valuable preoperatively for risk assessment, therapeutic decision making and surgical planning. The integrated volume rendering of brain surface topography, cortical veins, structural lesion, and sites of functional activation is also useful intraoperatively for defining cortical resection boundaries in patients with lesions in critical areas. PMID- 8726445 TI - Magnetic resonance imaging therapy. Intraoperative MR imaging. AB - The field of image-guided neurosurgery has developed under the premise that integrating the enhanced discrimination powers of CT scan and MR imaging into the operating room improves the accuracy, safety and precision of neurosurgery, and expand the realm of surgically treatable lesions. Through cooperation between General Electric Corporation and the Brigham and Women's Hospital, and open configuration MR suite was designed and implemented to attain these goals through real-time, intraoperative MR imaging in a true surgical suite. This system allows intraoperative acquisition of MR images without moving the patient, online image guided stereotaxy without preoperative imaging, and "real-time" tracking of instruments in the operative field registered to the MR images. The design and implementation of neurosurgery in the open-configuration MR imaging suite are summarized in this article. PMID- 8726446 TI - Superovulation. AB - Development of a superovulation technique that is successful, safe, and commercially available would revolutionize the equine breeding industry. However, the reality is that ovulation rates for mares following existing superovulatory treatment are much lower than for cattle. This dichotomy has been attributed to the relatively limited area available in the ovulation fossa for ovulation to occur, combined with the large size of the equine preovulatory follicle. In addition, the number of ovulations in the mare may be limited physiologically by the size of the follicular cohort that may be rescued by administration of gonadotropins. Clearly, additional research effort is necessary to optimize superovulation treatment regimens in the mare. PMID- 8726447 TI - Transvaginal aspiration. AB - This article describes in detail the procedures for collection of equine oocytes using a transvaginal ultrasound probe. Success in obtaining oocytes from humans, bovines, and horses are presented. The effect of repeated follicular aspiration of both cattle and horses is reviewed. PMID- 8726448 TI - Maturation and fertilization of equine oocytes. AB - Equine oocytes obtained either by transvaginal ultrasound-guided follicular aspiration or from slaughterhouse ovaries can be matured in vitro. This generally requires culture in TCM-199 containing serum and hormones for 30 to 36 hours. With this protocol, approximately 50% to 60% of the oocytes are at metaphase-II at the end of the culture period. At least some of these oocytes appear viable based on production of fertilized eggs either through in vitro fertilization or fertilization in vivo of a recipient mare. The success of producing equine embryos in vitro is still extremely low. More than likely the conditions for in vitro oocyte maturation are not optimized, and the techniques for capacitating equine spermatozoa are not adequate. The stallion sperm would appear more difficult to capacitate and many of the approaches used in other species have not worked in horses. To date, the only fertilization that has occurred with in vitro matured oocytes has been with sperm treatments containing the calcium ionophore A23187. Increased success with in vitro production of equine embryos may be gained through the use of assisted reproductive techniques such as partial zona dissection or intracytoplasmic sperm injection. PMID- 8726449 TI - Gamete intrafallopian transfer. AB - GIFT involves placement of a donor's oocyte into a surrogate's oviduct. Fertilization and embryo development occur within the recipient's reproductive tract. GIFT provides a viable method to obtain pregnancies from subfertile mares for which embryo transfer has been nonproductive. Currently, pregnancy rates after GIFT have been variable, although high success rates have been reported recently. Further refinement of techniques should allow GIFT to be used in research and commercial programs. PMID- 8726450 TI - Early embryonic development and evaluation of equine embryo viability. AB - Tremendous progress has been made in the development of assisted reproductive techniques that may enhance the reproductive efficiency of the horse. However, techniques that involve the manipulation of oocytes and/or embryos may themselves be detrimental to embryo viability and subsequent development. Therefore, an objective method of assessing viability of embryos before and/or after oocyte/embryo manipulation is desirable. At this time, morphologic evaluation is the most widely used method of determining the viability of equine embryos. Although morphologic assessment of embryo quality will not always accurately predict the survival of individual embryos, it is very useful for predicting the survival of groups of embryos. Other tests that have been used to evaluate equine embryo viability include (1) development during culture in vitro; (2) quantitating metabolism of the fluorescent substrate fluorescein diacetate; (3) quantitating uptake of the fluorescent stain DAPI; and (4) quantitating embryo metabolism. Although these tests offer potential advantages over morphologic assessment alone, their current limitations have prevented their routine use for embryo evaluation. It is likely that as improvements are made in these evaluation methods, they may offer advantages for use alone or in combination to more accurately assess the viability of equine embryos. PMID- 8726451 TI - Cryopreservation of equine embryos. AB - Principles and procedures for cryopreservation of equine embryos are described. Embryos less than 250 microM in diameter can be cryopreserved successfully if glycerol is used as the cryoprotectant. Cooling is takes place in such a way that most of the water leaves the cells before intracellular ice forms, and glycerol is removed after thawing without undue osmotic swelling of cells. Vitrification procedures also show promise for small embryos. Satisfactory procedures for cryopreserving embryos of more than 250 microM in diameter are not yet available. PMID- 8726452 TI - Use of semen as biopsy material for assessment of health status of the stallion reproductive tract. AB - Conventional light microscopic evaluation does not fully utilize potential indicators in seminal ejaculates for diagnosis of disorders of the reproductive tract. The technique of evaluation of all cellular components of semen, as described in this article, utilizing both light and transmission electron microscopy is a valuable diagnostic tool. Compare with other common biopsy procedures, use of semen as biopsy material is noninvasive, more representative than excisional biopsy, less expensive, and helps in the longitudinal evaluation after a therapeutic regimen. PMID- 8726453 TI - Methods for induction of capacitation and the acrosome reaction of stallion spermatozoa. AB - Methodologies to capacitate bovine spermatozoa, induce the acrosome reaction, and fertilize bovine oocytes in vitro have been established. The capability to do the same with stallion spermatozoa, however, is not available. Several different methods have been used to capacitate stallion spermatozoa with variable results. More basic research needs to be done to establish in vitro conditions necessary to capacitate and induce an acrosome reaction in stallion spermatozoa. Although much progress can be expected in this area, it is unlikely that the general practitioner will use these technologies in the near future. PMID- 8726454 TI - Analysis of stallion semen and its relation to fertility. AB - This article describes some of the basic methodology for conducting stallion semen evaluations. Not all of these assays will likely be conducted on every semen sample collected. Routine evaluations should include determination of semen volume, sperm concentration, and an estimation of the percentage of progressively motile sperm, at a minimum. Other assays can be used if a seminal problem is discerned at the beginning of the breeding season or after a stallion has recovered from an illness or injury. Additionally, laboratory assays are particularly important to conduct on cryopreserved spermatozoa, because these cells have been exposed to substantial temperature and osmotic insults (see Cryopreservation of Stallion Spermatozoa, this issue). The percentage of fully functional sperm is reduced during cryopreservation and the longevity of the cells surviving freezing is shortened. Therefore, to maintain high fertilization rates using cryopreserved sperm, only sperm samples containing sufficient numbers of high-quality cells should be used. Unfortunately, no single laboratory assay can estimate the fertilizing potential of a semen sample. What these analyses can do is discern semen samples that are likely to have poor fertilizing capacity. PMID- 8726455 TI - Cryopreservation of stallion spermatozoa. AB - The main advantage to using frozen semen in any breeding program is faster genetic gain for the inherited trait desired. Milk production of dairy cows doubled (from 26,000 to 52,000 kg of milk/cow per year) between 1950 and 1980, because the dairy industry was using semen only from bulls with the greatest genetic potential for milk production. This genetic gain could have been achieved without the use of frozen semen; however, the time required to achieve that same genetic progress would have been lengthened exceedingly. Fertility rates using frozen stallion spermatozoa are not equal to that of fresh semen, but with careful management of both the stallion and the mare fertility rates of 40% to 60% can be achieved for a single mare reproductive cycle and fertility rates of 60% to 75% during the course of a breeding season can be expected for a number of stallions. Because pregnancy rates have increased and because many breed registries now condone the use of frozen stallion semen, more people in the equine industry are using frozen semen. It has been the attempt of this article to make the practitioner aware of some of the problems that can occur when processing, freezing, storing, and using frozen semen, and how to control those problems. The practitioner also should be aware that there is large variation in semen quality from stallions, and that spermatozoa from some of the problem stallions can be improved by altering a "standard protocol," but that semen from some stallions will not cryopreserve effectively, regardless of the efforts put forth. PMID- 8726456 TI - GnRH therapy for subfertile stallions. AB - Poor fertility has been associated with hormonal imbalances in the stallion. The primary cause of this imbalance is unknown, but it may be related to abnormal gonadotropin secretion. Secretion of GnRH from the hypothalamus causes the release of LH and FSH from the anterior pituitary and these gonadotropins help maintain adequate steroid levels so that normal spermatogenesis can occur. Hypogonadotropic hypogonadism has not been documented in the stallion; however, GnRH therapy has improved the fertility of a limited number of horses. Downregulation of the HPT axis readily occurs in most other species treated with GnRH; however, the horse seems to be unusually resistant to the development of refractoriness. Although GnRH therapy is unlikely to be beneficial to most stallions suffering from fertility problems, it is not likely to be harmful. Complete endocrine and semen evaluations should be performed on all stallions that are being considered candidates for GnRH therapy. Sequential evaluations also are necessary to monitor the stallions response to therapy. Establishment of a database containing these parameters would improve greatly our ability to diagnose and select stallions that would have the best chance of responding to GnRH therapy. Development and use of universal standards for equine gonadotropins would be the single most important advancement in equine reproductive endocrinology because as this would allow accurate and meaningful comparisons to be made between the results obtained at numerous laboratories. PMID- 8726457 TI - Use of transvaginal ultrasound-guided puncture for elimination of equine pregnancies. AB - Twinning is a major cause of abortion in mares. Although early management of twins is quite successful, twins that remain after 40 days are difficult to manage. This article reviews the current knowledge of the use of transvaginal ultrasound probe for elimination of twins. Several techniques are presented, as well as discussion on the best time during gestation for this technique to be used. PMID- 8726458 TI - The Walter and Eliza Hall Institute of Medical Research. PMID- 8726459 TI - IBC sixth annual meeting on antibody engineering Dec 4-6 1995, La Jolla, CA. PMID- 8726460 TI - Alzheimer amyloid-beta peptide forms denaturant-resistant complex with type epsilon 3 but not type epsilon 4 isoform of native apolipoprotein E. AB - BACKGROUND: The apolipoprotein E (apoE) type epsilon 4 isoform specifies increased cerebral and cerebrovascular accumulation of amyloid-beta protein (A beta) and contributes to the genetic susceptibility underlying a large proportion (approximately 60%) of typical, sporadic Alzheimer disease. Unfortunately, in vitro biochemical studies of direct apoE isoform-specific interactions with A beta have been inconsistent, perhaps due to the use by different research groups of apoE isoform preparations in different conformational states (purified denatured versus native). MATERIALS AND METHODS: In the current study, we have investigated the possibility that synthetic A beta(1-40) preferentially associates with native apoE of either the type epsilon 3 or the type epsilon 4 isoform. RESULTS: Here, we demonstrate the preferential association of synthetic A beta(1-40) with native apoE epsilon 3. The complex between apoE epsilon 3 and A beta(1-40) could not be disrupted by sodium dodecyl sulfate. In a parallel assay, no denaturant-resistant association of A beta(1-40) with apoE epsilon 4 was detectable. CONCLUSIONS: These results support the notion that the apoE epsilon 4 isoform may foster beta-amyloidogenesis because apoE epsilon 4 is inefficient in forming complexes with A beta. PMID- 8726461 TI - Quantitative analysis of peptides from myelin basic protein binding to the MHC class II protein, I-Au, which confers susceptibility to experimental allergic encephalomyelitis. AB - BACKGROUND: An important issue in autoimmune diseases mediated by T cells, such as experimental allergic encephalomyelitis (EAE), is the affinity of the disease inducing determinants for MHC class II proteins. Tolerance, either due to clonal deletion or anergy induction, is thought to require high-affinity interactions between peptides and MHC molecules. Low-affinity binding is compatible with the hypothesis that breaking tolerance to self proteins does not have to occur for onset of disease. In contrast, a high-affinity interaction implies that an event leading to a breakdown of tolerance is central to the autoimmune process. MATERIALS AND METHODS: Detergent-solubilized and affinity-purified I-Au was incubated with varying concentrations of a set of peptides from myelin basic protein and a biotinylated peptide agonist. The specific complexes were separated from excess peptide by capture on antibody-coated plates, and the affinity of the peptides was measured by adding europium-labeled streptavidin and measuring the resultant fluorescence. RESULTS: The immunodominant and encephalitogenic determinant, Ac 1-11, was shown to bind to I-Au relatively poorly (IC50 = 100 microM), demonstrating that in this protein, immunodominance did not correlate with high-affinity binding. In contrast with the natural sequence, the ability of shorter analogs to induce EAE did correlate with their apparent affinity. CONCLUSIONS: The dominance of the natural determinant does not arise from a high affinity interaction with the MHC class II molecule. This suggests that other mechanisms are operative and that the specific T cell for this peptide/MHC ligand is of high affinity. PMID- 8726462 TI - H-RYK, an unusual receptor kinase: isolation and analysis of expression in ovarian cancer. AB - BACKGROUND: Protein tyrosine kinases play an important role in cellular metabolism as key components of signal transduction pathways. They are involved in cellular growth, differentiation, and development. Receptor tyrosine kinases (EGF receptor and c-erbB2) have been shown to be important in the pathogenesis of cancer. In ovarian cancer, overexpression of c-erbB2, a type I receptor, has been correlated with an adverse effect on survival of patients. MATERIAL AND METHODS: An unusual receptor tyrosine kinase, H-RYK, has been isolated from a complimentary DNA library of SKOV-3, an epithelial ovarian cancer cell line, using a polymerase chain reaction-mediated approach. RESULTS: The primary structure of the predicted amino acid sequence of the protein shows a novel NH2 terminal region. The catalytic region shows homology to other tyrosine kinases, the closest homology being with v-sea (39%). A significant alteration in the catalytic domain is that the highly conserved "DFG" triplet in subdomain VII is altered to "DNA." The gene was mapped to chromosome 3q22. A single transcript of 3.0 kb is expressed in heart, brain, lung, placenta, liver, muscle, kidney, and pancreas by Northern analysis with maximal expression in skeletal muscle. In situ hybridization analysis on human tissues demonstrated localization of message in the epithelial and stromal compartment of tissues such as brain, lung, colon, kidney, and breast. There was minimal to absent expression of H-RYK on surface epithelium of ovaries. In benign (3) and borderline tumors of the ovary (5), there was expression in the stromal compartment. However, in malignant tumors (24) there was increased expression predominantly confined to the epithelium. Polyclonal antisera raised against synthetic peptides recognize a 100-kD protein in ovarian cancer cells and other cell lines. In contrast to other receptor tyrosine kinases, the receptor did not phosphorylate in an in vitro kinase assay. CONCLUSIONS: The expression of this unusual receptor tyrosine kinase in epithelial ovarian cancer suggests that it may be involved in tumor progression, which needs further investigation. PMID- 8726463 TI - Parathyroid hormone-related protein is induced during lethal endotoxemia and contributes to endotoxin-induced mortality in rodents. AB - BACKGROUND: Parathyroid hormone-related protein (PTHrP) is a ubiquitous and highly conserved vasoactive peptide whose role and regulation in normal physiology remain an enigma. Recently, we demonstrated that low-dose endotoxin (LPS) induces intrasplenic, but not systemic, levels of PTHrP; and that tumor necrosis factor, a pro-inflammatory cytokine, is the major mediator of this effect. We have therefore hypothesized that, with higher, lethal doses of endotoxin, PTHrP could be induced in multiple tissues to such a degree that it could contribute to the lethality of septic shock. MATERIALS AND METHODS: Northern blot analysis was used to measure PTHrP mRNA levels in vital organs of rats after administration of a near lethal dose (5 mg/250 g) of LPS (or vehicle alone). Plasma levels of PTHrP were also measured by immunoradiometric assay. The ability of the immunoglobulin fraction of two different PTHrP(1-34) antisera to protect from LPS-induced lethality was also studied in mice using survival analysis. RESULTS: In response to a near-lethal dose of endotoxin, PTHrP mRNA levels increased acutely in every vital organ examined (spleen, lung, heart, kidney, and liver). Circulating levels of PTHrP also increased, peaking 2 hr after administration of high-dose endotoxin. Passive immunization of mice with anti-PTHrP(1-34) antibody 6 hr prior to administration of a lethal dose of LPS protected mice from endotoxin-induced death (p < 0.00005). CONCLUSIONS: These results suggest that PTHrP belongs to the cascade of pro-inflammatory cytokines induced during lethal endotoxemia that is responsible for the toxic effects of LPS. PMID- 8726464 TI - Vascular gene transfer of the human inducible nitric oxide synthase: characterization of activity and effects on myointimal hyperplasia. AB - BACKGROUND: Nitric oxide (NO) has been shown to decrease myointimal hyperplasia in injured blood vessels. We hypothesize inducible No synthase (iNOS) gene transfer even at low efficiency will provide adequate local no production to achieve this goal. MATERIALS AND METHODS: A retroviral vector containing the human iNOS cDNA (DFGiNOS) was used to transfer the iNOS gene into vascular cells and isolated blood vessels to answer the following questions: can vascular endothelial and smooth muscle cells support iNOS activity and will low efficiency iNOS gene transfer suppress myointimal hyperplasia in injured porcine arteries? RESULTS: DFGiNOS-infected sheep pulmonary artery endothelial cells (SPAEC) expressed significant iNOS mRNA and protein, releasing nitrite levels of 155.0 +/ 10.7 nmol/mg protein/24 h vs. 5.5 +/- 1.1 by control cells. Transduced rat smooth muscle cells (RSMC) also expressed abundant iNOS mRNA and protein, but, in contrast to SPAEC, NO synthesis was dependent on exogenous tetrahydrobiopterin (BH4) (291.8 +/- 10.4 nmol nitrite/mg protein/24 hr with BH4, 37.7 +/- 2.6 without BH4). Only porcine arteries infected with DFGiNOS following balloon injury exhibited a 3-fold increase in total NO synthesis and a 15-fold increase in cGMP levels over control vessels in a BH4 dependent fashion, despite only a 1% gene transfer efficiency. Transfer of iNOS completely prevented the 53% increase in myointimal thickness induced by balloon catheter injury; the administration of a NOS inhibitor reversed this effect. CONCLUSIONS: These in vitro findings suggest that vascular iNOS gene transfer may be feasible. Furthermore, a low gene transfer efficiency may be sufficient to inhibit myointimal hyperplasia following arterial balloon injury, although a source of BH4 may be required. PMID- 8726465 TI - Characterization and functional analysis of the human inducible nitric oxide synthase gene promoter. AB - BACKGROUND: Nitric oxide has a wide variety of homeostatic and pathological effects. Control of the production of nitric oxide by the inducible form of the enzyme resides in the 5' promoter region of the gene. Although control of the murine isoform has been investigated, little is known about the functional aspects of the human analog. MATERIALS AND METHODS: A 3.9-kb 5' nontranslated region of the human gene was cloned, sequenced, and several reporter constructs prepared. The promoter-reporter constructs were transfected into human or murine monocytoid cells and reporter expression quantified following cytokine activation of the cells. The production of nitric oxide was also monitored. RESULTS: Although a murine promoter-reporter functioned efficiently in both human and mouse cells, the human constructs functioned only in human cells. The activity of the mouse construct increased progressively with the addition of activating cytokines, but the human promoter-reporter did not. Although interleukin 1 beta drove expression of the human inducible nitric oxide synthase reporter, actual expression of nitric oxide required both interleukin 1 beta and interferon-gamma. CONCLUSIONS: The data indicate that despite the significant homology between the human and mouse inducible nitric oxide synthase promoter sequence, control of the two genes is quite different. In addition to being more efficient in promoter activity, the murine promoter responds increasingly to cytokines that are not effective for the human analog. It is also apparent that human inducible nitric oxide synthase is controlled at both the level of transcription and post translationally. PMID- 8726466 TI - Inhibition of hemozoin formation in Plasmodium falciparum trophozoite extracts by heme analogs: possible implication in the resistance to malaria conferred by the beta-thalassemia trait. AB - BACKGROUND: Human falciparum malaria, caused by the intracellular protozoa Plasmodium falciparum, results in 1-2 million deaths per year. P. falciparum digests host erythrocyte hemoglobin within its food vacuole, resulting in the release of potentially toxic free heme. A parasite-specific heme polymerization activity detoxifies the free heme by cross-linking the heme monomers to form hemozoin or malaria pigment. This biochemical process is the target of the widely successful antimalarial drug chloroquine, which is rapidly losing its effectiveness due to the spread of chloroquine resistance. We have shown that chloroquine resistance is not due to changes in the overall catalytic activity of heme polymerization or its chloroquine sensitivity. Therefore, the heme polymerization activity remains a potential target for novel antimalarials. In this study, we investigated the ability of heme analogs to inhibit heme polymerization and parasite growth in erythrocytes. MATERIALS AND METHODS: Incorporation of radioactive hemin substrate into an insoluble hemozoin pellet was used to determine heme polymerization. Incorporation of radioactive hypoxanthine into the nucleic acid of dividing parasites was used to determine the effects of heme analogs on parasite growth. Microscopic and biochemical measurements were made to determine the extent of heme analog entry into infected erythrocytes. RESULTS: The heme analogs tin protoporphyrin IX (SnPP), zinc protoporphyrin IX (ZnPP), and zinc deuteroporphyrin IX, 2,4 bisglycol (ZnBG) inhibited polymerization at micromolar concentrations (ZnPP << SnPP < ZnBG). However, they did not inhibit parasite growth since they failed to gain access to the site of polymerization, the parasite's food vacuole. Finally, we observed high ZnPP levels in erythrocytes from two patients with beta-thalassemia trait, which may inhibit heme polymerization. CONCLUSIONS: The heme analogs tested were able to inhibit hemozoin formation in Plasmodium falciparum trophozite extracts. The increased ZnPP levels found in thalassemic erythrocytes suggest that these may contribute, at least in part, to the observed antimalarial protection conferred by the beta-thalassemia trait. This finding may lead to the development of new forms of antimalarial therapy. PMID- 8726467 TI - Correction or transfer of immunodeficiency due to TNF-LT alpha deletion by bone marrow transplantation. AB - BACKGROUND: Mice with inactivated tumor necrosis factor (TNF) and lymphotoxin alpha (LT alpha) genes have profound abnormalities of the immune system including lymphocytosis, lack of lymph nodes, undifferentiated spleen, hypoimmunoglobulinaemia, and defective Ig class switch. Here, we asked whether this phenotype is due to incompetent lymphohemopoietic progenitors or to a defective environment. MATERIALS AND METHODS: Lethally irradiated TNF-LT alpha deficient and wild-type mice received bone marrow cells from either TNF-LT alpha deficient or wild-type mice. The reconstitution and transfer of the phenotype was followed by morphological and functional analyses. RESULTS: Bone marrow cells from wild-type mice restored the synthesis of TNF and LT alpha, corrected the splenic microarchitecture, normalized the lymphocyte counts in the circulation, and repopulated the lamina propria with IgA-producing plasma cells of TNF-LT alpha-deficient mice. Furthermore, the formation of germinal centers in the spleen and the defective Ig class switch in response to a T-cell dependent antigen was corrected, while no lymph nodes were formed. Conversely, the TNF-LT alpha phenotype could be transferred to wild-type mice by bone marrow transplantation after lethal irradiation. CONCLUSIONS: These data demonstrate that most TNF- and LT alpha-producing cells are bone marrow derived and radiosensitive, and that the immunodeficiency due to TNF-LT alpha deletion can be corrected to a large extent by normal bone marrow cell transplantation. The genotype of the donor bone marrow cells determines the functional and structural phenotype of the TNF-LT alpha-deficient adult murine host, with the exception of lymph node formation. These findings may have therapeutic implications for the restoration of genetically defined immunodeficiencies in humans. PMID- 8726468 TI - Effects of sulphydryl reagents on receptor-mediated hormonal responses at the cellular level: insulin-mimicking characteristics of thiol-blocking compounds in rat hepatocyte primary cultures. AB - The interference of various SH-blocking chemicals with the insulin-controlled regulation of the hepatic carbohydrate metabolism was studied in rat hepatocyte primary cultures. The organic mercurials PCMB, PCMBS, mersalyl the disulphide agents DTP, CPDS, disulfiram and the SH-alkylating reagent NEM were used as experimental SH-blocking model compounds. All studied compounds, except for NEM, induced an increased glycogen deposition comparable with the physiological insulin-induced glycogen-deposition. PCMBS appeared to be the most effective insulin-mimicking anabolic trigger. The action of the insulin molecule itself was potentiated by PCMBS as well as demonstrated by increased glycogen deposition, induced pyruvate kinase (PK) and decreased phosphoenol-pyruvate carboxykinase (PEP-CK) activity. However, cell-exposure to insulin and PCMBS in relatively high doses was destructive, as demonstrated by decreased glycogen levels, most probably as a result of insulin-receptor overstimulation and metabolic stress. Thus, SH-blocking compounds are able to trigger insulin-dependent metabolic processes. The relatively non-permeant organic mercurial PCMBS proved to be the most effective insulin-mimicking SH-blocking compound. PMID- 8726469 TI - Analysis of multiple conductance states observed in Ca2+ release channel of sarcoplasmic reticulum. AB - Both slowly and rapidly fluctuating currents were observed after incorporating a heavy sarcoplasmic reticulum (HSR) vesicle or a purified ryanodine receptor (RyR) of rabbit skeletal muscle into a lipid bilayer. The responses to the cis side Ca2+, ruthenium red and ryanodine confirmed that both were the currents through one Ca2+ release channel. From the analysis of conductance states, RyR is supposed to have two different configurations. The current through one configuration has a high reversal potential (HRP) (-18 mV) and that through another one has a low reversal potential (LRP) (-11 mV). The current with HRP fluctuates rapidly but the current with LRP always fluctuates slowly. From the analysis of both rapidly and slowly fluctuating currents, it was shown that there are four pores in RyR. Among the conductance states of the slowly fluctuating current with LRP, the conductance of the 4OP (four opened pores) state was smaller than four times that of 1OP (one opened pore). To explain this fact, we proposed a model of the channel, a large central pore in series with four small parallel pores. The calculation according to the model has a good fit with the bilayer measurement. PMID- 8726470 TI - Further investigation of some inhibitors on myogenic differentiation: mechanism of inhibition with HMBA on quail myoblasts transformed with Rous sarcoma virus. AB - To investigate the mechanism of myogenic differentiation, we are using quail myoblast cells (QM cells) transformed with a temperature-sensitive mutant of Rous sarcoma virus (ts-RSV), termed QM-RSV cells. At 35.5 degrees C, a permissive temperature for RSV, QM-RSV cells repeatedly proliferate without differentiation, but, at 41 degrees C, a nonpermissive temperature, myogenic differentiation proceeds. This temperature dependency of the differentiation is derived from protein kinase activity of pp60v-src, as tyrosine dephosphorylation is necessary for myogenic differentiation of QM-RSV cells. In this study, it was demonstrated that among four fusion inhibitors, aspirin, doxorubicin, HMBA and TPA, three of the inhibitors, except for TPA, inhibited myogenin gene expression. Moreover, HMBA inhibited myoblast fusion accompanying inhibition of tyrosine dephosphorylation of certain proteins, and recovered the tyrosine kinase activity of pp60v-src to a certain extent. To study the effect of HMBA on the intracellular localization of pp60v-src, detergent-soluble and detergent resistant fractions were prepared with Triton X-100. As a result, it was shown that pp60v-src mainly exists in detergent-resistant fraction at 35.5 degrees C. While almost all of the pp60v-src at 41 degrees C exists in detergent-soluble fraction. HMBA treatment retained pp60v-src in detergent-resistant fraction even at 41 degrees C. These results suggest that HMBA inhibits myogenic differentiation of QM-RSV cells by affecting the regulation of pp60v-src. PMID- 8726471 TI - Mutually exclusive distribution of the focal adhesion associated proteins and the erythrocyte membrane skeleton proteins in the human fibroblast plasma membrane undercoat. AB - The ends of stress fibers in the basal portion of cells (basal stress fibers) are anchored to focal adhesions, and stress fibers in the apical part of cells (apical stress fibers) are attached to the apical membrane, forming a structure (the apical plaque; KATOH, K. et al. (1995). Cell Motil. Cytoskel., 31: 177-195) resembling the focal adhesion. In addition to these two sites, stress fibers also make lateral contact with the plasma membrane but little is known about the molecular composition of this type of stress fiber-membrane interaction sites. Several actin-membrane association types are known, each employing a different set of proteins, and the focal adhesion and the erythrocyte membrane skeleton are the best characterized systems. We investigated by immunofluorescence microscopy if there is any morphological basis for the involvement of the erythrocyte membrane proteins in the stress fiber-plasma membrane association sites in cultured human fibroblasts. Our results indicated that fodrin (nonerythrocyte type spectrin) and ankyrin were generally associated with the plasma membrane, but that they were clearly excluded from the focal adhesion, the apical plaque and the stress fiber. Thus, it appears that the spectrin and the integrin based actin-membrane association systems are mutually excluded in the fibroblast membrane undercoat. Protein localization at the lateral stress fiber-membrane association site was also studied. Our data indicated that, while talin, vinculin, paxillin, fibronectin receptor and integrin beta 1 were present at the three stress fiber-membrane association sites, vitronectin receptor and integrin alpha v were absent from the apical plaque and the lateral association site. While the plasma membrane at the focal adhesion adheres tightly to a solid substrate, the cell surface of the apical plaque is free. Although the lateral association site faces the substrate, the molecular composition of this site is similar to that of the apical plaque. PMID- 8726472 TI - Protease-sensitive component(s) on the cell surface prevents self-fusion in a bisexual strain of Dictyostelium discoideum. AB - The sexual cycle of the cellular slime mold Dictyostelium discoideum offers a suitable system to analyze the mechanism of cell recognition during mating. Sexual cell fusion in D. discoideum typically occurs between complementary heterothallic strains. In addition, several bisexual strains are known which undergo sexual cell fusion with heterothallic strains of either mating type, but cannot do so by themselves. In the present study, trypsin digestion of cell surface molecules was found to induce self-fusion in a bisexual strain WS2162, suggesting the presence on the cell surface of a self-recognition molecule whose homophilic interaction interferes with the cell fusion mechanism. PMID- 8726473 TI - Pathway of C6-NBD-Ceramide on the host cell infected with Toxoplasma gondii. AB - Fluorescence microscopy, with dyes analog of ceramide, and transmission electron microscopy, were used to analyze lipid traffic during interaction of Toxoplasma gondii with host cells. It is C6-NBD-Ceramide (C6-NBD-Cer), a fluorescent analog of ceramide, stained the Golgi complex where was metabolized into fluorescent sphingolipid and glucosylceramide, and translocated via the Golgi complex to the plasma membrane of living cells. In uninfected cells, C6-NBD-Cer initially concentrated at the perinuclear region, and after its fluorescent products were present in the cytoplasm and the plasma membrane. In infected cells, the probe initially is stained the Golgi complex. After 4 hours incubation with C6-NBD-Cer, the parasites within the parasitophorous vacuole began to be stained, and at 5 hours incubation, the parasites are completely fluorescent. The Golgi complex, as revealed by fluorescent probe and electron microscopy, maintained its perinuclear position throughout the evolution of intracellular parasitism. These results suggested that the intracellular parasite used the lipid pathway of the host cell. PMID- 8726474 TI - Apoptosis of human embryonal carcinoma cells with in vitro differentiation. AB - An in vitro model of apoptosis and differentiation in human embryonal carcinoma (EC) cells was developed to study the mode of cell death and mechanisms of cell death in early development. Death of these cells was induced by treatment with retinoic acid (RA) under the same conditions as those for induction of differentiation. The manner of this cell death was apoptosis, not necrosis, with the morphological criterion for apoptosis. serum deprivation likewise caused apoptosis in both undifferentiated and differentiated EC cells. In differentiated EC cells, DNA fragmentation was observed in a smear pattern lacking the ladder pattern typically associated with apoptosis. However, in differentiated EC cells, DNA fragmentation occurred in various sizes. The expression of a carbohydrate antigen, LeY, a reported marker of apoptotic cancer cells, was increased by the treatment with RA. However, two-color analysis by flow cytometry with nick end labelling method revealed that LeY expression was closely correlated with cellular differentiation but not apoptosis after RA treatment in the human EC cell system. Collection of LeY positive cells by the magnetic bead method demonstrated that this expression was not due to apoptosis but rather to differentiation. On the other hand, LeY expression associated with apoptosis was induced by serum deprivation in both undifferentiated and differentiated EC cells. These results show that a subpopulation of undifferentiated EC cells takes the apoptotic pathway by induction of differentiation. The results also suggest that the population of cells taking an apoptotic pathway differs from a population of cells taking a differentiation pathway. This in vitro system for apoptosis in human EC cells will be useful for studies concerning apoptosis or programmed cell death in human developmental biology. PMID- 8726475 TI - Identification and characterization of a 230-kDa Golgi-associated protein recognized by autoantibodies from a patient with HBV hepatitis. AB - A serum from a patient with HBV hepatitis was found to contain autoantibodies reacting with various mammalian cells. Immunofluorescence staining of cultured cells with the autoantibodies revealed that the antigen was localized at perinuclear regions, where the Golgi markers alpha-mannosidase II and beta-COP were colocalized. The autoantigen disappeared from the perinuclear regions upon incubation with the fungal metabolite brefeldin A, and the immunostainable structures were fragmented into vesicles by treatment with nocodazole. These results strongly indicate that the antigen is localized at the Golgi complex. Immunoblots of cell lysates showed that the autoantibodies recognized a single protein with a molecular mass of 230 kDa in a variety of cell lines, indicating that the 230-kDa antigen is a conserved protein among mammalian species. We designated this protein GCP230 (Golgi complex-associated protein with a molecular mass of 230 kDa). when a postnuclear fraction was prepared and centrifuged, GCP230 was recovered in both cytosol and membrane fractions. Peripheral interaction of GCP230 with membranes was confirmed by phase separation in Triton X-114 solution and by extraction with sodium carbonate. Taken together, these results indicate that GCP230 is a peripheral membrane protein of the Golgi derived from the cytosol, although its function is not known at present. PMID- 8726476 TI - Involvement of active cellular mechanisms on the disorganization of oral apparatus in amicronucleate cells in Tetrahymena thermophila. AB - Ciliated protozoa, a group of unicellular eukaryotes, have two kinds of nuclei, a macronucleus (somatic nucleus) and a micronucleus (germinal nucleus) in a single cell. We previously reported that amicronucleate cells of Tetrahymena thermophila induced by nocodazole gradually lost their oral apparatus (OA) and ciliary rows but that amacronucleate cells did not. Since the macronucleus is responsible for the gene expression in the vegetative phase, the effects of actinomycin D and cycloheximide on the disorganization of the OA in amicronucleate cells induced by nocodazole were investigated. These inhibitors prevented the disorganization of the OA in amicronucleate cells. Amicronucleate cells did not grow even in the medium supplemented with high concentration of Fe, Cu and folinic acid which allow cells to grow without formation of food vacuoles. The results suggest that the macronucleus in the amicronucleate cells plays an active role in the induction of disorganization of the OA and malfunctions of nutrient uptake from the cell surface and/or in the fundamental cell division mechanisms, resulting in the death of amicronucleate cells. PMID- 8726477 TI - Influence of air exposure treatment on alveolar type II epithelial cells cultured on extracellular matrix. AB - We cultured isolated alveolar type II epithelial cells on a collagen gel matrix. At confluence, cultured type II cells were exposed to air. Under these conditions, the cellular density of the type II cells increased and they nodularly aggregated. The cultured cells consisted mainly of flattened epithelia intermingled with cuboidal cells. In the cytoplasm and at the apical surface of cuboidal cells, a surfactant protein was detected by immunohistochemistry and electron microscopy. Electron microscopic examination revealed that the surfactants in the cytoplasm increased when the cells were exposed to air. On the other hand, the flattened cells morphologically resembled type I cells in vivo. Air exposure treatment on the collagen gel matrix promoted the maintenance of the characteristic differentiation of alveolar epithelial cells. This culture system seemed to provide an appropriate physiological environment in which to study differentiation and disorders of pulmonary alveoli. PMID- 8726478 TI - Comparison between the distribution of 7H6 tight junction-associated antigen and occludin during the development of chick intestine. AB - 7H6 antigen is a novel tight junction-associated protein first identified in our laboratory with a monoclonal antibody generated by immunizing mice with a bile canaliculus-rich membrane fraction obtained from adult rat liver. Although the antigen is found preferentially at the tight junction of epithelial cells in various organs in adult mammals, the relationship between 7H6 antigen and other tight junction proteins remains obscure. In the present study we examined the relationship between the immunohistochemical distribution of 7H6 antigen and occludin in chick intestinal epithelial cells during development in vivo, since occludin is the only known membrane protein composing tight junctions which is found thus far exclusively in avian tissues. 7H6 antigen was found as coarse dots along the basolateral cell membrane of intestinal epithelium in the chick embryo, whereas it was found as a fine dashed line along the cell border in the chicken. In contrast, occludin was found linearly together with ZO-1 at the cell border of intestinal epithelia both in the chick embryo and chicken. The possible significance of such different localization is briefly discussed. PMID- 8726479 TI - Discrimination by any other name. PMID- 8726480 TI - Managed care: III. Whose business is patient care? PMID- 8726481 TI - Usefulness of physical examinations in the psychiatric emergency service. PMID- 8726482 TI - Characteristics of psychiatric patients. PMID- 8726483 TI - "Echoes from a dungeon cell": a doctor's view of his incarceration. PMID- 8726484 TI - Impact of external versus internal case managers on hospital utilization. PMID- 8726485 TI - The Internet: applications for mental health clinicians in clinical settings, training, and research. PMID- 8726486 TI - Depression among aging persons with schizophrenia who live in the community. AB - OBJECTIVES: The study used the social antecedent model of depression in later life proposed by Linda K. George to develop a multifactorial model of predictors of depression among aging persons with schizophrenia living in the community. METHODS: The SHORT-CARE and the Network Analysis Profile, along with supplemental items, were administered to 117 outpatients age 55 and older who received DSM-III R diagnoses of schizophrenia before age 45. Bivariate and multivariate analyses examined associations between 27 variables derived from George's model and the presence of depression. A cutoff score for the depression scale of the SHORT-CARE was used to detect clinically significant depression. RESULTS: Forty-four percent of the sample had probable pervasive depression. Bivariate analysis indicated that 16 variables were significantly associated with depression, and logistic regression showed that six variables were strong predictors. Depression was associated with the presence of positive symptoms, physical limitations interfering with activities, younger age, diminished linkage of members of the social network, lower income, and a smaller proportion of social network members who provided sustenance such as food and money. An 11-variable model correctly classified 81 percent of cases. CONCLUSIONS: The level of depression among older outpatients with schizophrenia is approximately one and a half to three times the level in the general older population. Depression in this patient group is associated with nonclinical variables that are independent of psychoses and that resemble patterns found among general older populations. Several of the variables are potentially alterable and therefore afford opportunities to enhance the well being of persons with schizophrenia who live in the community. PMID- 8726488 TI - Psychiatric evaluation and psychotherapy in the patient's second language. AB - Use of a patient's second language in psychiatric evaluation and treatment has a variety of effects. Patients frequently undergo psychiatric evaluation in their second language, yet competence in a second language varies depending on the phase of illness. Evaluation of bilingual patients should ideally be done in both their languages, preferably by a bilingual clinician or by a monolingual clinician with the help of an interpreter trained in mental health issues. Cultural nuances may be encoded in language in ways that are not readily conveyed in translation, even when the patient uses equivalent words in the second language. The monolingual clinician may clarify these nuances through consultation with a clinician who shares the patient's first language and culture or with an interpreter. In psychotherapy, patients may use a second language as a form of resistance, to avoid intense affect. Therapists may use language switching to overcome this resistance and to decrease emotional intensity, if necessary. Psychotherapy can also be affected by the attitudes toward speaking that are part of the patient's culture. Discussions with bilingual and bicultural consultants can elucidate these effects for the therapist who is unfamiliar with the patient's culture. PMID- 8726487 TI - Variation among VA hospitals in length of stay for treatment of depression. AB - OBJECTIVE: The purpose of the study was to determine the degree of interhospital variation in length of stay for patients treated for depressive disorders in Department of Veterans Affairs (VA) medical centers and to identify the number of hospitals with significantly longer or shorter than average lengths of stay (outlier hospitals). METHODS: The sample included 6,374 veterans discharged from acute psychiatric inpatient units at 107 medical centers in 1992 with a primary diagnosis of major depressive disorder or depressive disorder not otherwise specified. To identify statistical outliers, the average length of stay at each medical center was compared with the overall sample mean while controlling for differences in case mix between hospitals. RESULTS: Patients' demographic characteristics, treatment history, and severity of illness all significantly predicted length of stay at the patient level. After case mix factors were controlled for statistically, approximately 29 percent of the medical centers were found to have mean lengths of stay significantly different (p < .01) from the sample mean. Specifically, 15 percent of the medical centers were short-stay outliers, and 14 percent were long-stay outliers. CONCLUSIONS: Despite the fact that the treatment regimen for depression is relatively standardized and all VA medical centers operate under the same administrative model and reimbursement system, a high degree of interhospital variation was found in treatment duration for depression. Results led to the tentative postulation that variations in treatment duration reflect differences in physicians' practice styles. Substantial opportunities may exist for reducing expenditures for the treatment of depression within the VA health care system. PMID- 8726489 TI - Social and living skills of new long-stay hospital patients and new long-term community patients. AB - OBJECTIVE: The study assessed differences in social behavior skills and everyday living skills in two groups of psychiatric patients: hospital inpatients with stays of between six months and five years (new long-stay patients) and discharged patients who had resided in the community for a similar length of time (new long-term patients). METHODS: In a cross-sectional study based on a point prevalence survey in an outer London psychiatric hospital, 23 new long-stay and 23 new long-term patients were rated using the Social Behaviour Scale (SBS) and Basic Everyday Living Skills (BELS). RESULTS: Mean scores on both scales indicated significantly poorer functioning in the new long-stay group. Ratings on the BELS self-care and domestic skills subscales also indicated poorer functioning in that group. No significant differences between groups were found for the subscales measuring community and social relations skills. Significant gender differences emerged in ratings of domestic skills; men's scores indicated more impairment in the new long-stay group than in the new long-term group, while women's scores were comparable between the groups. CONCLUSIONS: Developing, enhancing, and maintaining skills among new long-stay patients, especially self care and domestic skills, may facilitate their discharge into supported community settings. PMID- 8726490 TI - Quality of care and use of less restrictive alternatives in the psychiatric emergency service. AB - OBJECTIVE: The study examined factors affecting clinicians' decisions in the psychiatric emergency service about referring patients to less restrictive alternatives to inpatient care. Indicators of quality of care and the severity of the patient's condition were a particular focus. METHODS: Trained mental health professionals observed the evaluations of 425 patients in seven California county general hospitals. Multivariate modeling was used to examine variables thought to predict disposition to alternative care. RESULTS: Less restrictive alternatives were available for 61 percent of the 425 patients and were used for 39 percent; they were overlooked by clinicians in 14 percent of cases and considered but not used in 8 percent. Patients' need for a controlled hospital setting, as indicated by the severity of their condition, was most important in determining use of hospital alternatives. Quality of care, especially the clinician's ability to engage patients in treatment at a level appropriate to their functioning, was also a significant predictor of whether alternative care was considered or used. CONCLUSIONS: Under managed care, clinicians are under extreme economic pressure to use less restrictive alternatives, thereby reducing costly inpatient care. To ensure quality of care in general hospital emergency services, the development of supervised hospital alternatives is crucial. Clinicians should be encouraged to engage patients in treatment if appropriate use of alternative care is a goal. PMID- 8726491 TI - Housing accommodation preferences of people with psychiatric disabilities. AB - OBJECTIVE: The study sought to identify the types of housing preferred by people with psychiatric disabilities seeking accommodations through a community mental health service in Sydney, Australia. METHODS: Housing options available in the mental health service's catchment area were identified, and the extent of demands on residents' behavior that was associated with each option was assessed using an established measure. Clients examined descriptions of each housing option and rank-ordered the options based on their preferences. Clients' current levels of functioning and symptoms were also assessed. RESULTS: Clients most preferred environments that ensured living alone in settings of low behavioral demand. Living in one's own home was the most preferred option, followed by living in government-subsidized housing, For-profit boarding houses were preferred over psychiatric group homes, and homelessness, long-term hospitalization, and crisis accommodations were least preferred. Shelter, privacy, food, and safety were highly valued housing attributes. Demographic characteristics, level of functioning, and severity of psychiatric symptoms did not predict accommodation preferences. CONCLUSION: Consumers' resistance to psychiatric group housing with high levels of behavioral demand is unrelated to consumer characteristics and should be considered in planning suitable accommodations for persons with psychiatric disabilities. PMID- 8726492 TI - Grief therapy for relatives of persons with serious mental illness. AB - Grief is a key reaction experienced by many relatives of persons with serious mental illness. Parents, spouses, and children may endure great longing for the quality of their former relationship with the ill person, as well as grief over dashed hopes and aspirations. The author describes a format for grief therapy for relatives of persons with serious mental illness that is based on four tasks of mourning: making the loss real, expressing overt and latent affect, accommodating to the loss through relationships other than that with the mentally ill person. Grief therapy is a potentially appropriate complement to existing approaches for family members, such as psychoeducation. PMID- 8726493 TI - Factors associated with multiple hospitalizations of patients with borderline personality disorder. AB - OBJECTIVE: The study sought to identify aspects of borderline personality disorder and comorbid axis I conditions associated with multiple hospitalizations in a sample of patients with borderline personality disorder. METHODS: Data were collected as part of a larger study of treatment course of inpatients with this disorder. Predictors of multiple hospitalization from three domains were considered: demographic characteristics, criteria of borderline personality disorder, and comorbid axis I pathology. A sequential model-building strategy was used, with each domain considered separately and result combined into an overall regression model of factors related to multiple hospitalization. RESULTS: Three factors were related to frequency of hospitalization: anorexia, psychotic symptoms, and suicidality. Other variables that are often assumed to be related to hospitalization, such as depression, did not emerge as significant predictors. CONCLUSIONS: Clinicians should intervene early to address the emergence of certain axis I conditions among patients with borderline personality disorder. Increased expenditure of outpatient resources may prevent hospitalization when these conditions are present. PMID- 8726494 TI - A national survey of hospital -and community-based approaches to pretrial mental health evaluations. AB - To document the scope of recent trends in performing pretrial mental health evaluations, telephone interviews were conducted with 157 authorities in 50 states and the District of Columbia. These authorities included forensic mental health program directors and forensic clinical professionals. In all states, evaluations were made on an outpatient basis to some extent, and 45 states used inpatient evaluations. However, only ten states relied primarily on inpatient pretrial evaluations, generally in facilities with statewide catchment areas. A total of 32 states relied primarily on outpatient evaluations, generally conducted by providers with regional or local catchment areas. PMID- 8726495 TI - Impact of a psychiatric outreach service for homeless persons with schizophrenia. AB - Since 1988 a 24-hour psychiatric out reach service has been in operation in the inner city of Sydney to provide services to residents of refuges for the homeless. A total of 506 homeless persons with schizophrenia were referred to the outreach service between April 1988 and mid-1992, of whom 91 failed to attend. Hospitalization data were collected for the four years before and the four years after each individual's referral to the service. After the introduction of the service, the rate and duration of psychiatric hospital admissions for residents with schizophrenia who were treated by the outreach service decreased significantly, whereas those who failed to attend showed no such decrease. PMID- 8726496 TI - Utility of the chemical use, abuse, and dependence scale in screening patients with severe mental illness. AB - The Chemical Use, Abuse, and Dependence (CUAD) Scale was administered with other scales to 100 consecutive admissions to a psychiatric hospital. Moderately high correlations with other measures of drug and alcohol use were obtained. Although more research with the CUAD is necessary, it may be potentially useful in screening for substance use among severely mentally ill patients. PMID- 8726497 TI - Suicidal inpatients' experience of constant observation. AB - Suicidal inpatients' perspectives on the experience of constant observation by staff or lay workers were sought through interviews using open-ended questions with 14 patients for whom constant observation ranged from 16 hours to 3.5 days. Interview transcripts were analyzed using phenomenological methods to determine common themes. The three major themes that emerged from participants' responses were physical protection, restoration of hope, and distressing incidents. Participants reported that constant observation was beneficial because it contributed to their physical safety and restored hope. They also reported that the absence of supportive interactions, frequent staff changes, and lack of privacy adversely affected their experience. PMID- 8726498 TI - Rural and urban differences in continuity of care after state hospital discharge. AB - In a study to determine differences in continuity of care for state hospital patients discharged to rural and urban areas, all community mental health centers in Virginia were asked to complete a brief questionnaire about each patient discharged to the centers in fiscal year 1992. Discharges to rural centers had significantly higher levels of continuity of care on four of five dimensions of continuity. The authors conjecture that rural centers' lower staff ratios and decreased role boundaries may make them better able than urban centers to carry out the diverse tasks needed to help discharged patients resume community living. PMID- 8726499 TI - A brief method of assessing adult inpatients' level of need for community support systems. AB - The study examined the usefulness of the Level of Community Support Systems (LOCSS) scale in rating psychiatric inpatients' need for community supports after discharge. The scale was compared with two other instruments, the Community Placement Questionnaire (CPQ) and the Community Supports and Services Checklist (CSSC) in assessing 105 inpatients. LOCSS scale ratings were related to service utilization indicators, to items on the CPQ measuring community functioning ability, and to the CSSC score. The LOCSS scale correctly classified the level of difficulty of community placement for 69 percent of patients, indicating its usefulness as a screening tool in planning community-based psychosocial rehabilitation services. PMID- 8726500 TI - Host factors in the pathogenesis of HIV disease. PMID- 8726501 TI - Questions about CD8+ anti-HIV lymphocytes in the control of HIV infection. PMID- 8726502 TI - Mucosal receptors and T- and B-cell immunity. PMID- 8726503 TI - HIV-1 genetic variability: implications for the development of HIV vaccines. PMID- 8726504 TI - V3 region genotyping of HIV isolates in northern Uganda: heteroduplex mobility assay, nucleotide sequence and phylogenetic analysis. Italian-Ugandan Cooperation AIDS Program. PMID- 8726505 TI - HIV-1 biological phenotype varies with severity of infection across HIV-1 genetic subtypes. PMID- 8726506 TI - Immunotyping HIV-1 in the 1990s. WHO Collaborating Network. PMID- 8726507 TI - Overview of HIV vaccine development. PMID- 8726508 TI - A recombinant HIV-1 virus-like particle vaccine: from concepts to a field study. PMID- 8726509 TI - Role of adjuvants in HIV vaccine design. PMID- 8726510 TI - Relative importance and functional role of genetic elements targeted for a live attenuated AIDS vaccine. PMID- 8726511 TI - Vaccination of mice and nonhuman primates using HIV-gene-containing DNA. PMID- 8726512 TI - Immune responses to the HIV rev regulatory gene. PMID- 8726513 TI - HIV-2 vaccine trials in cynomolgus monkeys. PMID- 8726514 TI - Vaccination of chimpanzees against HIV-1. PMID- 8726515 TI - Evaluation of protective immunity to HIV-1 in human PBL-SCID mice. PMID- 8726516 TI - Vaccination of cynomolgus monkeys with whole inactivated or live-attenuated simian immunodeficiency virus. PMID- 8726517 TI - Establishment of WHO-sponsored field sites for HIV vaccine evaluation in developing countries. PMID- 8726518 TI - Combination strategies for vaccination against HIV. PMID- 8726520 TI - Preparation for HIV vaccine efficacy trials. PMID- 8726521 TI - Strategies for the achievement of prophylactic vaccination against HIV. PMID- 8726519 TI - Immunological aspects of patients with HIV-1 disease following immunization with recombinant gp160 (VaxSyn). PMID- 8726522 TI - Functional studies of bispecific antibodies directed against HIV-1 and the Fc gamma I receptor type I. PMID- 8726523 TI - Conformation of the HIV-1 gp 120 envelope glycoprotein. PMID- 8726524 TI - Some novel approaches to the control of HIV and select aspects of HIV pathogenesis. PMID- 8726525 TI - TAR decoys and trans-dominant gag mutant for HIV-1 gene therapy. PMID- 8726526 TI - Genetic pharmacomodulation of nucleoside analogues and gene therapy for AIDS and cancer. PMID- 8726527 TI - The HIV-1 regulatory genes tat and rev as targets for gene therapy. PMID- 8726528 TI - A replication-deficient human immunodeficiency virus-1 genome as an interference inducing provirus. PMID- 8726529 TI - Development of ribozyme gene therapy for HIV infection. PMID- 8726530 TI - Immune reconstitution for HIV disease. PMID- 8726531 TI - The neurotopography of vowels as mirrored by evoked magnetic field measurements. AB - The auditory evoked neuromagnetic field elicited by synthetic specimens of the vowels [a], [ae], [u], and [i] was recorded over the left and the right hemisphere of 11 subjects. The N100m and the SF deflection of the recorded signal was submitted to equivalent current source analysis using the model of a single dipole in a spherical volume conductor. Vowel processing was hypothesized to occur in a multistage process rendering a sequence of representations of the acoustic input. Vowel representations were considered to differ among each other in the features they make salient, thus, in the kind of dissimilarity relationship they establish, and, by implication, in terms of the vowel space defined by the respective set of dissimilarities. It was investigated whether a mapping exists between at least one of a number of hypothetical vowel spaces and the cortical response space spanned by the spatial distribution of vowel evoked equivalent current dipoles. Although the spatial configuration of vowel evoked sources proved to be highly variable across subjects, the ordering of distances between N100m and SF equivalent current dipole locations turned out to correspond to the ordering of distances between the corners of a vowel trapezium. There were some, albeit weak, indications of hemispheric differences in vowel processing. The results suggest that the spatial distribution of the equivalent current dipole sources of both the N100m and the SF deflection of the neuromagnetic field elicited by vowels reflect a processing stage transitional between auditory and phonetic representation. PMID- 8726532 TI - Microstates in language-related brain potential maps show noun-verb differences. AB - Brain processing of grammatical word class was studied analyzing event-related potential (ERP) brain fields. Normal subjects observed a randomized sequence of single German nouns and verbs on a computer screen, while 20-channel ERP field map series were recorded separately for both word classes. Spatial microstate analysis was applied, based on the observation that series of ERP maps consist of epochs of quasi-stable map landscapes and based on the rationale that different map landscapes must have been generated by different neural generators and thus suggest different brain functions. Space-oriented segmentation of the mean map series identified nine successive, different functional microstates, i.e., steps of brain information processing characterized by quasi-stable map landscapes. In the microstate from 116 to 172 msec, noun-related maps differed significantly from verb-related maps along the left-right axis. The results indicate that different neural populations represent different grammatical word classes in language processing, in agreement with clinical observations. This word class differentiation as revealed by the spatial-temporal organization of neural activity occurred at a time after word input compatible with speed of reading. PMID- 8726533 TI - Optic aphasia: a case with spared action naming and associated disorders. AB - AG, a pure case of optic anomia (object naming impaired; action naming good) is described. We consider the fit of experimental data from AG to different theoretical accounts of optic aphasia. Overall, we find no evidence for impairments intrinsic to semantic representations, but we note a number of problems that we interpret as indicating a slight, and specific, weakness in semantic access from vision. We also note a mild problem in generating names to a cue (verbal fluency). The main aim of the report was to provide a full description of tests of visual, semantic, and speech output skills in this patient in relation to the processing of visually presented objects and scenes, within a sequential information processing account which enables some theoretical implications to be drawn, albeit not conclusively. PMID- 8726534 TI - Agraphia in Alzheimer's disease: an independent lexical impairment. AB - This study was conducted to delineate the pattern of the writing impairments in 12 patients with Alzheimer type dementia. The patients performed writing tasks involving regular and irregular words and nonwords given by dictation as well as a decision test composed of printed words and pictures requiring phonologic, lexical, and semantic processing. Writing from dictation demonstrated a predominant, but nonisolated, lexical deficit. In order to better evaluate this lexical disorder, the correlation between the dictation writing scores and the decision task scores was analyzed. No significant correlation was found among scores for irregular words, phonologically plausible errors, and scores of the lexical or semantic decision tasks, but there was a significant correlation among scores for the nonwords, nonphonological spelling errors, and scores of the phonologic decision task using printed words. These results would suggest that the "lexical" deficit in agraphia, i.e., difficulty in retrieval or loss of the spelling representations of words, is independent of the lexical or semantic capacities involved in other modalities. PMID- 8726536 TI - Educational and gender normative data for the Boston Naming Test in a group of older adults. AB - The 60-item version of the Boston Naming Test (BNT) was administered to a group of 176 normal older adults in Middle Tennessee ranging in age from 60 to 93 years. Results are reported in five age groupings by gender and education. Although a few studies have previously examined the performance of the elderly on the BNT, the current study attempted to recruit individuals in proportion to the demographics of the regional population including gender, race, and, as much as possible, education and occupation. The results indicate that while age significantly affected confrontational naming ability, the interaction of age and education appeared to be a better predictor of performance. As with previous studies, more variability was found in higher age groups and in those with less education. Naming ability remained stable in the higher education group (> or = 12th grade) until 80 years, whereas those patients with less than a high school education demonstrated a decrement in naming ability at 70 years. Current results also revealed a gender bias for this population, with males scoring higher on 17 of 19 items where a significant difference existed. PMID- 8726537 TI - Production and perception of word tones (pitch accents) in patients with left and right hemisphere damage. AB - The present paper addresses the question of the functional lateralization of tones in tone languages. Tonal perception and production of right-hemisphere damaged (RHD) and left-hemisphere-damaged (LHD) speakers of East Norwegian were investigated. East Norwegian is a tone language with an opposition between two tones (pitch accents). The ability to distinguish auditorily between the two accents was normal in the RHD group but reduced in the LHD group. Tonal production was near normal in the RHD group, whereas the LHD group tended to have a production deficit. PMID- 8726535 TI - Cerebral mechanisms for suppression of inappropriate information during sentence comprehension. AB - In two experiments we investigated the extent to which interference from contextually inappropriate information was attenuated or suppressed over time in the two cerebral hemispheres during sentence comprehension. Subjects viewed centrally presented sentences ending in either a homophone or a homograph and made speeded judgments as to whether a laterally presented test word was related to the overall meaning of the sentence. Suppression of contextually inappropriate forms of homophones was found when test words were presented to either hemifield, but suppression of inappropriate senses of homographs was found only when test words were presented to the right visual hemifield. The results from the homograph experiment are consistent with the hypothesis that right and left hemisphere semantic selection systems operate in qualitatively different ways. The results from the homophone experiment suggest that while the left hemisphere may be more efficient at suppression, both hemispheres possess the ability to suppress inappropriate information to some degree. PMID- 8726538 TI - Intranasal apomorphine rescue therapy for parkinsonian "off" periods. AB - Eleven patients with levodopa-related motor fluctuations were scored before and after intranasal apomorphine monotherapy, and the motor responses were compared with those with levodopa/carbidopa in this openlabel study. Oral trimethobenzamide was used to prevent apomorphine-induced nausea. Three measures of motor performance were employed: (a) the Unified Parkinson's Disease Rating Scale (UPDRS) motor battery; (b) a timed hand-tapping test; and (c) the Webster's step-seconds test. The magnitude of the motor-score improvement after apomorphine administration was very similar to that after the usual doses of levodopa/carbidopa in the 10 patients completing the study; this was true for all three outcome measures. A major advantage of apomorphine was the rapid onset of clinical response, which typically occurred in < 10 min, as well as the ease of administration. Major side effects, beyond those experienced with levodopa/carbidopa, were limited to nausea and vomiting (three patients) and orthostatic hypotension (one patient); however, only a single patient dropped out of the study as a consequence. These results indicate that intranasal apomorphine is effective in rapidly relieving parkinsonian "off" states and that, for most patients, trimethobenzamide is an effective and well-tolerated antiemetic for use with apomorphine. PMID- 8726539 TI - Adjunctive cabergoline therapy of Parkinson's disease: comparison with placebo and assessment of dose responses and duration of effect. AB - Adjunctive cabergoline or placebo, in doses up to 5 mg daily, were administered to Parkinson's disease patients with short-duration levodopa responses in a 6 month double-blind trial. The 13 patients randomized to cabergoline and completing the study had significantly improved Unified Parkinson's Disease Rating Scale (UPDRS) motor scores and timed hand-tapping test scores. Serial measurements on test days documented improved scores: (a) before the first levodopa (and cabergoline) dose of the day, (b) at the time of the peak levodopa effect, and (c) at the end of the levodopa response cycle, 5 h after test doses. Continued testing verified that these therapeutic responses were sustained for at least 48 h after the last cabergoline dose. Patients randomized to placebo failed to improve on any of these measures. In a subsequent open-label dose-escalation phase, further improvement was documented as the dosage was gradually raised to 10 mg daily. As in the double-blind phase, levodopa reduction allowed the improvement to occur in the absence of significantly increased dyskinesias. Other side effects were more substantial with higher doses, however, including two of 11 patients with hallucinations and confusion. In summary, adjunctive single daily-dose cabergoline therapy resulted in long-lasting, dose-related improvement in parkinsonism not seen in patients receiving placebo. PMID- 8726540 TI - Lack of relation between genetic polymorphism of cytochrome P-450IID6 and sporadic idiopathic Parkinson's disease. AB - We investigated genetic polymorphism of the cytochrome P-450 CYP2D6 gene from white patients with idiopathic Parkinson's disease (IPD). The mutations of the CYP2D6 gene associated with the poor metabolizer (PM) phenotype of the debrisoquine/sparteine polymorphism were analyzed in DNA of 130 IPD patients by a polymerase chain reaction (PCR)-based DNA amplification combined with Xba I restriction fragment length polymorphism (RFLP) analysis in 105 patients. Another mutation located in exon 6 was analyzed by Hha I RFLP in 94 IPD patients. The frequencies of the different CYP2D6 gene mutations were compared to the frequencies in sex- and age-matched white control population with chronic bronchitis. The rate of genotypically defined PM and the frequencies of the different mutations were not significantly different in IPD patients and controls. These results fail to confirm the previously reported results concerning CYP2D6 gene mutations in IPD. These equivocal results might be related to methodologic problems. However, other hypotheses have been suggested: impairment of neuronal CYP 2D6 expression, transient modification of CYP 2D6 phenotype, or linkage of CYP2D6 gene to the candidate gene locus directly involved in IPD. PMID- 8726541 TI - Effect of one month's treatment with peripherally acting catechol-O methyltransferase inhibitor, entacapone, on pharmacokinetics and motor response to levodopa in advanced parkinsonian patients. AB - Twelve parkinsonian patients with levodopa-related end-of-dose fluctuations in disability were studied in an open-label trial to examine the effects of peripheral catechol-O-methyltransferase (COMT) inhibition with entacapone on pharmacokinetics and metabolism of levodopa and on clinical response to levodopa after a single dose and after 4 weeks' medication with entacapone. The clinical response was assessed with continuous monitoring using the motor part of Unified Parkinson's Disease Rating Scale. Entacapone increased statistically significantly the mean area under the plasma concentration-time curve (AUC) of levodopa by 29% after a single dose and by 21% after 4 weeks' administration, without affecting other pharmacokinetic parameters of levodopa. The AUC of 3-O methyldopa decreased by 45% and AUC of homovanillic acid by 21% after 4 weeks' dosing with entacapone. The duration of motor response to levodopa increased significantly from 2.3 h to 3.2 h (i.e., by 39%) after a single dose and to 3.4 h (i.e., by 48%) after 4 weeks' medication with entacapone. The magnitude of clinical response remained unchanged, but peak latency of motor response was prolonged after 4 weeks' medication. The duration and magnitude of dyskinesias also increased. Peripheral COMT inhibition with entacapone increased significantly the bioavailability of levodopa and prolonged its antiparkinsonian effect after a single dose and after repeated dosing for 4 weeks. Thus entacapone seems to be a valuable adjuvant to levodopa treatment in parkinsonian patients with end-of-dose failure. PMID- 8726542 TI - Ropinirole in the treatment of levodopa-induced motor fluctuations in patients with Parkinson's disease. AB - Forty-six patients with Parkinson's disease experiencing motor fluctuations and not optimally controlled on levodopa received as adjunct therapy a new nonergoline dopamine agonist, ropinirole, in a 3-month randomized placebo controlled trial. Ropinirole significantly reduced the duration of off periods as assessed by self-scoring diary cards. There were more nonserious dopaminergic adverse events in the ropinirole group. More patients withdrew because of adverse events or insufficient therapeutic effect in the placebo group. Ropinirole has beneficial adjuvant effects in parkinsonian patients with moderate motor disability and motor fluctuations. PMID- 8726543 TI - Role of dopamine mesolimbic system in opioid action of psychotropic analgesic nitrous oxide in alcohol and drug withdrawal. AB - Psychotropic analgesic nitrous oxide (PAN) has been used successfully in the treatment of alcohol and drug withdrawal in > 15,000 cases. It is an opioid and thus the first gaseous member of the opioid family. We propose the existence of two mutually antagonistic opioid systems as underlying addictive withdrawal states; mu and kappa. PAN as a multipotent opioid activates these systems. Dopamine (DA) activity in the nucleus accumbens appears to be controlled by kappa and mu-receptors, with mu enhancing and kappa inhibiting release. In morphine and alcohol withdrawal, there is severe inhibition of dopamine release from nucleus accumbens. We thus infer that a probable major therapeutic effect of PAN is in modulating this dopamine system, thereby correcting the severe deficit in dopamine release found in withdrawal states. This has been achieved without any transfer of addiction to PAN in any of the treated patients because of modulation of DA in the nucleus accumbens by PAN. This effect may also explain its anticraving action. PMID- 8726544 TI - Involvement of the endogenous opioid system in the drinking behavior of schizophrenic patients displaying self-induced water intoxication: a double-blind controlled study with naloxone. AB - Previously we found significant suppression of polydipsia in a schizophrenic patient with PIP syndrome (psychosis, intermittent hyponatremia, and polydipsia). Suppression was obtained with a small dose of naloxone injected once every 2 weeks in long-term repeated studies. We attempted to confirm the effect of naloxone on PIP syndrome by using a double-blind controlled study. The body weights of eight schizophrenic inpatients with PIP syndrome were checked five times daily, and the maximum weight gain during 1 day was chosen as an index of their polydipsia. Naloxone (0.6 mg in three divided doses) or placebo (saline) injection was given once every 2 weeks three times. Assignment to either the naloxone or placebo series was done randomly in a double-blind, crossover design. Naloxone decreased the maximum weight gain per day significantly in five cases. However, naloxone also increased weight gain significantly in three cases. There was no correlation of the weight-increasing effect of naloxone with the duration and intensity of excessive drinking. Our findings showed that the endogenous opioid system might be related to compulsive drinking behavior in the PIP syndrome and that opioid antagonists such as naloxone or naltrexone could be useful in the therapy of PIP syndrome. PMID- 8726545 TI - Permanent blindness after cyclosporine neurotoxicity in a kidney-pancreas transplant recipient. AB - Blindness is an extremely rare complication of cyclosporine neurotoxicity. In all 10 cases in the literature, this form of blindness is completely reversible with the reduction or withdrawal of cyclosporine. We describe the first case of sudden, complete, and permanent blindness within 36 h after administration of intravenous cyclosporine in a kidney-pancreas transplant recipient. PMID- 8726546 TI - Significantly higher plasma haloperidol level during cotreatment with carbamazepine may herald cardiac change. AB - We investigated the effect of carbamazepine (CBZ) cotreatment with haloperidol (HP) on cardiac change in Japanese psychiatric patients without heart disease. Of 21 patients cotreated with CBZ, 11 showed lengthening of QT interval corrected for heart rate (QTc) (> 440, mean percentage change, 29), of whom two patients developed congestive heart failure because of left ventricular dysfunction. Especially in these two patients, plasma HP levels were significantly higher than speculated from the correlations. It is possible to speculate that, especially on CBZ cotreatment, significantly higher HP plasma level with QTc prolongation may herald heart failure. PMID- 8726547 TI - Abnormalities of antioxidant metabolism in a case of Friedreich's disease. AB - We report a patient with Friedreich's disease (FD) who exhibited abnormalities of antioxidant metabolism, including decreased levels of glutathione peroxidase, glutathione reductase, and selenium, and an increased lipid peroxide index. These abnormalities became normal after treatment with N-acetylcysteine, selenium, and low-dose vitamin E therapy. Treatment was associated with a decreased rate of clinical decline. FD is a neurodegenerative disorder that may be related to disturbed antioxidant metabolism; the disorder may be treatable with antioxidant compounds. PMID- 8726548 TI - Smooth pursuit eye movements in the evaluation of famotidine adjunctive therapy of schizophrenia: a preliminary report. AB - Smooth pursuit eye movements (SPEM) are often abnormal in schizophrenic patients and have been proposed as a trait marker of the disorder. We explored the use of SPEM as an outcome measure in an open-label clinical trial of famotidine, an H-2 antagonist, in patients with schizophrenia; famotidine has been proposed as an adjunctive medication, particularly for negative symptoms. Prior studies using SPEM as an outcome measure have not found a significant effect with "typical" neuroleptic medication, and one study found greater SPEM dysfunction with clozapine treatment. In this study, 19 schizophrenic subjects were stabilized for at least 1 week on conventional neuroleptic medications and then administered oral famotidine, 100 mg daily, for an additional 3 weeks. SPEM and clinical measures were assessed. Whereas Brief Psychiatric Rating Scale (BPRS) and Schedule for Assessment of Negative Symptoms (SANS) scores decreased significantly with famotidine administration, there was no significant change in SPEM performance over the course of the study. Two subjects (11%) doubled their signal/noise ratio and maintained this increase after famotidine discontinuation, whereas three subjects (17%) approximately halved this ratio and returned to baseline after famotidine discontinuation. SPEM changes were not found to correlate significantly with changes in BPRS or SANS scores. These findings suggest that SPEM dysfunction reflects a "trait" rather than clinical "state" in schizophrenia, and changes in SPEM performance might not be expected always to parallel changes in clinical ratings. PMID- 8726549 TI - Partner-specific sexual practices among heterosexual men and women with multiple partners: results from the French national survey, ACSF. Analyse des Comportements Sexuel en France. AB - Heterosexual men and women with several partners are at risk of acquiring and transmitting sexually transmitted diseases and HIV. Risk depends on parameters such as the sexual practices themselves which may vary according to the type of partner (regular vs. casual). It is therefore important to describe the sexual practices and identify the correlations between the type of partner and these practices among heterosexuals with multiple partners. A subsample of all subjects having had at least two sexual partners during the previous year (n = 1644) was obtained from the ACSF survey (n = 20,055), the French national telephone survey on sexual behavior conducted between September 1991 and February 1992. Questions concerned in particular sexual practices of the last encounter as well as type of partner. Petting and vaginal penetration were almost systematic, mutual manual stimulation and orogenital sex were common, while self-masturbation and anal sex were infrequent. On average, a condom was seldom used. However, it was used more often when the partner was occasional. Nonpenetrative and oral practices were also more frequent with occasional partners. Women tended to report lower frequencies of practices and of condom use than men. A subset of heterosexuals with multiple partners engaged in safe sex. Practices tend to be partner specific, with safer sex practices more likely to occur with occasional partners, although the magnitude of the difference is moderate. PMID- 8726550 TI - Assessment of sexual beliefs and information in aging couples with sexual dysfunction. AB - The incidence and prevalence of sexual dysfunction indicate that a large portion of the U.S. population will experience a sexual dysfunction, with older individuals being particularly at risk. Cognitive factors (e.g., attitudes, misinformation, beliefs) may contribute to the development of sexual dysfunction or influence response to treatments. The purpose of the current study was to evaluate the psychometric properties of a brief instrument designed to assess information and beliefs regarding sexual functioning in an aging population. Results indicate that the Sexual Beliefs and Information Questionnaire (SBIQ) had adequate test-retest reliability and is an internally consistent measure of sexual knowledge and beliefs. MOst individuals had adequate knowledge regarding sexual functioning as indicated by correct responses to most items; however, there were several items missed more often than expected. Exploratory factor analysis suggests an underlying five-factor structure of the SBIQ. Scores were not correlated with years of education, mood, or marital satisfaction. Scores on the SBIQ improved after a brief standardized educational intervention. The SBIQ is a simple and psychometrically sound measure of sexual knowledge and myths that can assist clinicians to identify and possibly rectify knowledge deficits. PMID- 8726551 TI - Premarital sexual standards among U.S. college students: comparison with Russian and Japanese students. AB - The study of the sexual permissiveness of young adults has been popular topic in sociology and social psychology, especially since the empirical and theoretical work of Reiss. We extended previous research on premarital sexual standards by examining the degree of sexual permissiveness and the endorsement of the traditional double standard in a large sample of young adults in the United States (N = 1043). In addition, comparative data were collected from young adults in two other countries: Russia (N = 401) and Japan (N = 223). American subjects expressed more acceptance of premarital sex than did the Russian and Japanese subjects. Men were more sexually permissive than women in the U.S. and in Russia but not in Japan. The degree to which the double standard was endorsed also depended on culture and gender. Russian subjects were more likely to endorse the double standard than Japanese and American subjects. However, American men were most likely to endorse the traditional double standard concerning sex early in the dating relationship. PMID- 8726552 TI - Sexual socialization and motives for intercourse among Norwegian adolescents. AB - The impact of gender differences in sexual socialization on early sexual experiences among Norwegian adolescents is discussed. The material comprises a stratified sample of 920 adolescents ages 16-20 years in a Norwegian county. Data were collected by means of questionnaires. Of the respondents, 55.5% were girls and 44.5% were boys. 52.3% of girls and 41.4% of the boys had coital experience. The most common reasons for having had the first sexual intercourse were being in love, curiosity or excitement, and sexual arousal. Findings from a discriminant analysis showed that emotional reasons were more important to girls, whereas boys seemed more practical in sexual matters. More boys than girls reported that the reason for having had their most recent intercourse was that the partner wanted it. This indicates that if girls do not want sex, boys seldom use pressure. Girls set the premises for sexual interaction but are not as sexually skilled as boys. PMID- 8726553 TI - Transsexualism--general outcome and prognostic factors: a five-year follow-up study of nineteen transsexuals in the process of changing sex. AB - Nineteen transsexuals, approved for sex reassignement, were followed-up after 5 years. Outcome was evaluated as changes in seven areas of social, psychological, and psychiatric functioning. At baseline the patients were evaluated according to axis I, II, V (DSM-III-R), SCID screen, SASB (Structural Analysis of Social Behavior), and DMT (Defense Mechanism Test). At follow-up all but 1 were treated with contrary sex hormones, 12 had completed sex reassignment surgery, and 3 females were waiting for phalloplasty. One male transsexual regretted the decision to change sex and had quit the process. Two transsexuals had still not had any surgery due to older age or ambivalence. Overall, 68% (n = 13) had improved in at least two areas of functioning. In 3 cases (16%) outcome were judged as unsatisfactory and one of those regarded sex change as a failure. Another 3 patients were mainly unchanged after 5 years. Female transsexuals had a slightly better outcome, especially concerning establishing and maintaining partnerships and improvement in socio-economic status compared to male transsexuals. Baseline factors associated with negative outcome (unchanged or worsened) were presence of a personality disorder and high number of fulfilled axis II criteria. SCID screen assessments had high prognostic power. Negative self-image, according to SASB, predicted a negative outcome, whereas DMT variables were not correlated to outcome. PMID- 8726554 TI - Continued lack of evidence for transmission of human immunodeficiency virus through vaginal intercourse: a reply to Carey and Kalichman. PMID- 8726555 TI - [The gestalt problem in neurobiology]. AB - The paper concerns the concept of gestalt in the framework of neuronal mechanisms. Two hypotheses are discussed: 1) hierarchy of neurons (gnostic units), 2) synchronization of neurons. It is concluded that gestalt is based on hierarchical vectorial organization of neurons. The high frequency oscillations in gamma range (40-200 Hz) are of intrinsic origin. They are operating as amplifiers of synaptic inputs of neurons involved in gestalt representation. PMID- 8726556 TI - [The cerebral basis of subjective experiences: the hypothesis of information synthesis]. AB - The hypothesis is advanced that the events of the subjective experience emerge as a result of the synthesis of three kinds of information in the cortical areas which are crucial for this mental function. The three kinds of information are presented by the sensory inputs, reproduction from memory, and that from motivational centers. The hypothesis is based on the studies of brain mechanisms of perception and thinking. It has been shown that sensation emerges as a result of the synthesis of information about the physical parameters and significance of the stimulus performed in the projection cortex neurons. This synthesis is provided by the circular run of nerve impulses from the projection cortex to associative cortex, then to hippocampus and the hypothalamic motivation centers with subsequent return to the projection cortex. It has been shown also that in thinking operations the cortical connections converge to definite centers named the interaction foci. Their topography is specific for particular thinking operations. Thus, in imaginative thinking the foci are located in the temporo parietal areas and in abstract verbal thinking in the frontal cortex. It is suggested that comparison and synthesis of information in the interaction foci result in decision making. PMID- 8726557 TI - [The dynamics of the dominant alpha-rhythm frequency in the perception and reproduction of time intervals]. AB - Dependence of the dominant alpha frequency on the processes of perception and subsequent reproduction of time intervals by rhythmic hand pressing was analysed. Two groups of subjects were separated, which demonstrated either an increase (I) or a decrease (II) in the dominant alpha frequency after the transition from perception to reproduction. In the I group 83% of the subjects reproduced time intervals with a delay. In the II group reproduction in advance was observed in 75% of the subjects. The dynamics of the dominant alpha frequency probably reflects formation of the optimal functional state underlying the individual level of activation and properties of the higher nervous activity in the subjects from both groups. PMID- 8726558 TI - [Sex and age differences in the subjective assessment of time by preschoolers]. AB - Time perception was studied in children under school age from 2.5 to 6 years. Using cross-modal matching, the children assessed durations of 1-, 3-, 5-, 7-, and 10-sec sound signals. It was found out that children under school age, irrespective of sex, more accurately estimated the extreme durations in the internal range (1 and 10 sec). In contrast to girls, boys reveal a clear-cut tendency to minimize the variability of assessments with age. Nevertheless, this tendency is not related with the attention level. PMID- 8726559 TI - [The reactivity of the sensorimotor rhythm in patients with depressive disorders]. AB - Reactions of mu-rhythm to sound (unspecific reactivity), actual and imaged movement (specific reactivity) were studied in 30 right-handed patients with psychogenic (Pg) and endogenous (Eg, cyclophrenia and low-progredient schizophrenia) subdepressive disorders and 23 healthy subjects. Predominant derangement of specific reactivity was revealed in patients which consisted in attenuation of reactions and reorganization of interhemispheric relations. During the actual movement there was a decrease in mu-rhythm reactivity in the left hemisphere and its increase in the right one. The imaged movement was associated with some decrease in the right-hemisphere mu-rhythm reactivity. The degree of reorganization of interhemispheric relations was higher in Eg patients, especially in schizophrenic ones. Abnormal ipsilateral reactions were induced by the actual (in Eg patients) or imaged movement (in Pg patients) with the right hand. These abnormal reactions are regarded as a return to an immature (childish) form. Only in schizophrenic patients the level of unspecific reactivity is lower than in the healthy subjects. There were no hemispheric differences in the sound induced mu-rhythm reaction both in patients and healthy subjects. PMID- 8726560 TI - [The disordered organization of the cortical processes in depression]. AB - The study of depressive patients with the method of the EEG mapping revealed some stable differences from the norm. Firstly, in depression there are the disturbances of the definite cortical zones' activation: the right anterior and the left posterior cortical parts are hyperactivated while the left anterior and the right posterior-- relatively inhibited, evidently causing the informational processing disturbances. The important link of these disturbances--the disconnection of the anterior and the posterior cortical regions that could be called "transversal functional blockade". The second feature of the electrical activity of the brain in depression is the absence of the high frequency component of alpha-rhythm, that evidently in normal conditions is connected with the positive emotions mechanisms. The third feature is the difference in arousal reaction in the alpha- and beta-bands frequencies. PMID- 8726561 TI - [The characteristics of the latent periods of movements in cats during the differentiation of tonal and amplitude-modulated signals with changes in the modulation from high frequencies to low]. AB - The latent periods of the forepaw movements of cats were studied in the course of differentiation task performance. Pure tones and amplitude-modulated stimuli had to be differentiated. It was found out that the mean latency of motor responses to pure tones was significantly higher than that of responses to modulated signals. The latencies to the applied stimuli did not differ in the simple task (without differentiation). The latencies were influenced not only by the absolute values of modulation frequency but also by the frequency shift in successive experiments. Lowering modulation frequency decreased the latency. This effect was especially expressed around the values of 205-207 and 33-34 Hz. PMID- 8726562 TI - [The distribution in real time of the spike train activity in the cortical neurons of cats during learning]. AB - Analysis of the dynamics of distribution of coincident impulses in pairs of neurons revealed modifications in the structure of neuronal interaction in the process of learning. In electrodefensive sound conditioning coincident paired impulses of the neurons in cat's motor cortex were timed to the moment of the conditioned stimulus presentation. In food-procuring time conditioning coincident impulses were concentrated around the moment of the reinforcement administration. The described temporal pattern of coincident impulse distribution is considered as a manifestation of a conditioned reaction at the level of neuronal interaction. This conclusion is confirmed by the finding that coincident impulses may be grouped within the period of expected conditioned reaction even in the absence of the defensive or food-procuring movement in well-trained cats. PMID- 8726563 TI - [Excess dietary arginine inhibits the development of the food-acquisition habit in white rats]. AB - Behavioural effects of NO precursor L-arginine (Arg) were studied. Arg was shown to impair place learning with food reinforcement in the maze task. Arg was introduced per os immediately after the training session, i.e., 24 hours prior to the next session in doses of 50, 100 and 250 mg/kg. Intake of Arg in the dose of 100 mg/kg induced the most effective learning inhibition. Special tests demonstrated that Arg in the applied doses did not influence food motivation. Analysis of Arg effects suggests that the observed behavioural changes are probably due to the excess NO synthesis from Arg in brain tissues which, in turn, may result in cell damage. PMID- 8726564 TI - [The role of the cholinergic system of the brain in the mechanisms of dissociated learning]. AB - The mechanisms of dissociated learning in rats were studied with application on conditioning technique and pharmacological analysis of memory. Anticholinesterase drugs were shown to be interchangeable in inducing dissociated states in learning. Anticholinergic substances completely neutralized the ability of cholinergic drugs that create dissociated states. It was shown that muscarinic cholinergic receptors largely contribute in inducing dissociated states. The findings well agree with the earlier author's hypothesis that the cholinergic brain system plays a decisive role in the mechanisms of dissociated learning. PMID- 8726565 TI - [Acoustically guided behavior in the early ontogeny of the long-eared owl: the development of hearing sensitivity]. AB - The development of hearing sensitivity was studied in 15 nestlings and 7 adult long eared owls by cochlear recording potentials. The dynamic of frequency threshold characteristics of cochlear microphonic, action potential of cochlear nerve, and electromyogram of reflectory contraction of middle ear muscle were followed from the 1st to 40 day posthatching. The upper limit of hearing determined by cochlear microphonic, was about 3 kHz on the 1st day after hatching, 7 kHz on the 6th day, and 10 kHz on the 11th. Within 11 days after hatching the low-frequency range of higher sensitivity was revealed (with the maximum at 0.5 kHz), which was the same as that of monotonic signals effective for stimulation of the feeding reaction. The middle frequency range of higher sensitivity corresponded to the spectrum of own vocalization of the nestlings. During the period of formation of the patterned vision there was a delay in development of low- and middle-frequency sensitivity of cochlear microphonic while the high-frequency thresholds continued to decrease. After this period the guiding stimulation for eliciting feeding reaction changes from the auditory for the visual. The action potential of the cochlear nerve could be recorded from the 6th day simultaneously with the appearance of locating head movements and the reflectory contraction of the middle ear muscle. PMID- 8726566 TI - [The direct activating influence of the lateral hypothalamic preoptic area on the thalamic synchronizing system]. AB - In order to reveal the mechanisms of participation of the preoptic hypothalamic area in sleep-wake cycle regulation the effects of its electrical stimulation on activity of the thalamic median center were studied in chronic experiments in cats. Ipsi- and contralateral projections from the lateral preoptic area to the thalamic median center were electrophysiologically demonstrated. Activatory influence of the preoptic area was shown on the thalamic mechanisms of spindle activity generation. The obtained data suggest the interaction of the preoptic area with the structures of non-specific thalamus via the direct pathways to be one of the mechanisms of the preoptic area participation in sleep initiation and formation of the slow-wave sleep. PMID- 8726567 TI - [The effect on rat behavior of phenamine contained in a cannula chronically implanted in the neostriatum]. AB - Avoidance conditioning in a shuttle-box was studied in rats under conditions of daily three-week amphetamine microinjections (45 mcg) into the rostral striatum and in rats with chronically implanted intrastriatal amphetamine-containing cannulae (45 mgg/0.75 mcl). Amphetamine stimulatory behavioural effects were recorded in both groups of animals (improvement of avoidance conditioning, stereotyped hyperactivity etc.). The dopaminomimetic effects in rats with implanted cannulae were less expressed than in those with daily injections. High chemical stability of cannulated amphetamine was demonstrated by means of HPLC technique. Neurodegenerative changes were described in the loci of amphetamine administration similar to those in its long-lasting systemic injections. The methodological problems are discussed which are essential for neuropharmacological studies of behaviour. PMID- 8726568 TI - [A comparison of the frequency of the Straub tail elevation reaction as a measure of the activity of the endogenous opiate system in rats of different strains]. AB - Stress-induced and exploratory activity-related Straub tail elevation was studied as a probable measure of endogenous opiate system activity in rats from catalepsy susceptible GC strain as well as PM+ and PM- strains bred from Wistar stock for the presence and absence of predisposition to a stereotyped hyperkinesis with pendulum-like movements. All the three strains were characterized by lower incidence of Straub tail elevation than in the control Wistar stock. Moreover, the PM+ strain demonstrated a lower incidence of the studied reaction than PM- the difference between the strains being manifested during exploratory activity but not in immobilization stress. PMID- 8726569 TI - [The characteristics of the brain serotonin system and anxiety in the C57BL and CBA mouse strains]. AB - Serotonin metabolism and 5-HT2A and 5-HT1A specific binding were studied in high aggressive C57BL and low-aggressive CBA mice. In three main tests for anxiety (elevated plus-maze "dark-light test", and social contacts) C57BL mice revealed higher anxiety than CBA, Activity of tryptophan hydroxylase, the key enzyme in serotonin biosynthesis was significantly lower in the midbrain and neostriatum of C57BL than CBA mice. The specific binding of [3H]-ketanserin in C57BL was higher in the frontal cortex and lower in the neostriatum than in the same structures of CBA mice being indicative of the differences in 5-HT2A receptor density. There were no significant differences in 5-HT1A receptor density (as indicated by specific [3H]-8-OH-DPAT binding) between the strains. It was suggested that decreased serotonin metabolism and characteristic distribution of 5-HT2A receptors can underlie the expression of genetic predisposition to anxiety and aggression. PMID- 8726570 TI - [The effect of changes in the dynamic equilibrium in the microtubular and microfilamentous systems on neuronal plastic reactions]. AB - Effects of disruption or stabilization of microfilaments or microtubules on formation of neuronal plastic reactions (NPR) were studied in isolated Lymnaea stagnalis neurons. Disruption of these cytoskeletal elements blocked the development of the NPR. After stabilization of the microtubules the dynamics of development and retention of the NPR became dependent on the stimulation series. Stabilization of the microfilaments blocked the development but improved the retention of the NPR. The role of the dynamic cytoskeletal reorganization in the process of NPR formation is discussed on the basis of the obtained data. PMID- 8726571 TI - [The relationship of the dynamics of the recovery of a manipulatory habit after unilateral neocortical damage in white rats with a limb preference and free choice]. AB - The choice of a forelimb by a rat within two months after the damage of the left motor cortex (the right motor cortex remaining intact) depended on the degree of recovery of the injured forelimb. There was no such correlation when the right hemisphere was damaged. Distinctions in the course of recovery after the left- and right-hemisphere injury were most pronounced in the ambidextrous rats. Under free choice conditions within two months after the injury the "handed" rats predominantly chose the initially preferred ("damaged") forelimb, while ambidextrous rats preferred to use both forelimbs. PMID- 8726572 TI - [The participation of serotonin S1A and S2 receptors in the formation of different levels of anxiety in male mice under the influence of the experience of social victories and defeats]. AB - Influence of repeated experience of victories or defeats in daily social confrontations on formation of different levels of anxiety was studied in male mice (winners and losers). Repeated experience of defeats in losers produced the pronounced anxiety evaluated in the plus-maze and in the partition test (by the low-expressed behavioural reaction to another conspecific). Flezinoxan (0.5 mg/kg, i. p. 30 min), 5-HT1A receptor agonist, produced changes in the partition test behaviour in winners and controls and was not effective in losers. Ritanserin (2.0 mg/kg, i. p. 30 min), 5-HT2 receptor antagonist, decreased reactivity to a conspecific in losers and winners and was not effective in control. It has been supposed that development of pronounced anxiety in losers is accompanied by a decrease in 5-HT1A receptor sensitivity. PMID- 8726573 TI - [The cerebral organization of verbal processes in adult subjects]. PMID- 8726574 TI - [The coordinated activity of functionally similar hypothalamic neurons under different motivational-emotional conditions]. PMID- 8726575 TI - [The effect of the blockade of the serotonin-modulating protein SMP-69 by antibodies on rat exploratory behavior]. PMID- 8726576 TI - [The numerousness judgement by the hooded crow of a set consisting of 12-25 elements]. PMID- 8726577 TI - [The effect of imipramine on the dynamics of forced swimming in rats following enucleation and removal of the epiphysis]. PMID- 8726578 TI - [A software-hardware package for studying the coordination of human eye, head and hand movements]. PMID- 8726580 TI - [A short psychophysiological dictionary]. PMID- 8726579 TI - [A method for the quantitative evaluation of search activity and refusal to search in an experiment with rats]. PMID- 8726582 TI - Analytical applications of liquid phase chemiluminescence reactions--a review. AB - This paper reviews the literature on analytical applications of quantitative liquid phase chemiluminescence (CL) from 1991 to mid-1995. Other relevant reviews in this general area are also cited to provide an historical perspective. The focus is on the two major analytical techniques used in conjunction with flow through CL detection, namely flow injection (FI) and liquid chromatography (LC). Entries have been tabulated under these two headings and are categorized in terms of the analyte, CL reaction, sample matrix and limits of detection. PMID- 8726585 TI - Bioluminescence and chemiluminescence literature. The 1995 literature: Part 3. PMID- 8726583 TI - Two processes responsible for chemiluminescence development in the course of iron mediated lipid peroxidation. AB - The kinetics of chemiluminescence (CL) accompanying Fe(2+)-induced lipid peroxidation (LPO) in liposome suspension has been investigated. A sequence of stages was observed, namely: (1) fast CL flash (FF); (2) latent period (LP); (3) slow CL flash (SF) and (4) stationary chemiluminescence (SL). The first three stages are known to reflect the Fe(2+)-mediated LPO process. In spite of the fact that at the stage of SL Fe2+ has completely oxidized and MDA has not accumulated, CL intensity was found to increase and after 0.5-1 h reached a value that was several times higher than SF amplitude. The maximal SL level was linearly dependent on the initial Fe2+ concentration and was not dependent on liposome concentration in the suspension. The nature of the processes responsible for CL emission at the stage of SL has been investigated using free radical reaction inhibitors and measurement of CL spectra. The SL spectrum was observed in the red region (lamda > 590 nm) in contrast to the SF CL spectrum (maximum at 540nm). SL amplitude was strongly inhibited by sodium azide (40%), superoxide dismutase (SOD) (30%), desferrioxamine and EDTA (30%), whereas mannitol, ethanol, alpha tocopherol and butylated hydroxytoluene were ineffective. The data obtained indicate that CL at the stage of SL is not directly related to LPO process, i.e. lipid free radical recombination. The mechanism of stationary CL generation is discussed. PMID- 8726584 TI - Myeloperoxidase-based chemiluminescence of polymorphonuclear leukocytes and monocytes. AB - Luminol and lucigenin chemiluminescence (CL) responses produced by separated human blood polymorphonuclear leukocytes (pmn) and monocytes (mono) have been studied following stimulation with the surface-receptor agonist fMLP (a synthetic chemotactic peptide) and the protein kinase C activator phorbol myristate acetate (PMA). Pmn produced two- to threefold the luminol CL and superoxide anion (O2-) levels of mono; lucigenin CL was similar for both cell-types. The myeloperoxidase (MPO) inhibitor salicylhydroxamic acid (SHA) abrogated luminol but not lucigenin CL in both cell types, but did not further inhibit the already grossly subnormal luminol CL responses seen with MPO-deficient cells which produced normal lucigenin CL. SHA also profoundly inhibited the luminol CL response in a cell free MPO-H2O2 system. Mono lucigenin CL does not appear to specifically measure O2- production. These data show that luminol CL provides a useful measure of pmn and also mono MPO activity. However, analysis of the effects of various reactive oxygen species (ROS) scavengers, assessed on phagocyte and cell-free CL systems (both MPO-H2O2 and superoxide generating) suggest that the luminol CL signal is not entirely dependent on MPO activity. PMID- 8726587 TI - New approaches to adjuvant chemotherapy for breast cancer. AB - Adjuvant chemotherapy improves the disease-free and overall survival of patients with resectable, early-stage breast cancer. However, the effect is modest, and we need means of increasing its impact. Retrospective and prospective studies on the effects of increased drug dosage have demonstrated that within some dose ranges, increasing the dose intensity (total drug dose divided by total treatment time) improves the outcome of treatment. Dose escalation to higher levels, sufficient to require growth factor and autologous stem cell support, is the subject of ongoing randomized study. Because there are both theoretical and practical limits on the potential effectiveness of single-cycle, high-dose chemotherapy, researchers are developing alternative means of increasing the effect of chemotherapy. One theoretically advantageous approach, predicted to be superior by kinetic models, is "dose-dense" chemotherapy administration. This approach consists of multiple cycles of escalated-dose chemotherapy administered at very short intervals. When administering presumed non-cross-resistant regimens or agents with overlapping toxicity, clinicians can increase dose density by using the sequential treatment plan. Furthermore, this plan can also facilitate the addition of new active drugs. PMID- 8726586 TI - Management of patients with breast cancer: from chemotherapy to stem cell support. Introduction. PMID- 8726588 TI - The emergence of peripheral blood progenitor cells to support intensive chemotherapy for patients with breast cancer. AB - Increasing evidence supports the hypothesis that "dose" is critical to the clinical outcomes of cytotoxic chemotherapy for patients with breast cancer. Clinical trials continue to investigate whether higher doses of chemotherapy lead to proportionate improvements in the outcomes of patients. Delivery of dose intensive chemotherapy has been facilitated by technological advancements in supportive care. Improved antiemetics have led to increased patient tolerance of the most acute symptoms of aggressive chemotherapeutic dosing. Chemotherapy induced myelosuppression may be minimized in a lineage-specific manner by appropriate use of hematopoietic cytokines such as filgrastim (granulocyte colony stimulating factor), sargramostim (granulocyte-macrophage colony-stimulating factor), and/or epoetin alfa (erythropoietin). However, cumulative myelotoxicity occurs with dose-intensive chemotherapy over multiple cycles despite adjunctive cytokine support. Additionally, no cytokine has yet been demonstrated to support platelet production to any clinically significant degree although several regulators of platelet production (such as thrombopoietin, IL-6, and IL-11) are in clinical trials. Many cytokines can induce the mobilization of hematopoietic progenitor and stem cells from the bone marrow into the circulating blood pool, where these cells may be harvested. Clinical use of these cytokine-mobilized peripheral blood progenitor cells (also known as PBPCs or, commonly, as blood stem cells) has documented the effectiveness of these cells to reconstitute multilineage blood production following very high-dose chemotherapy. The ease with which PBPCs can be collected and their reproducible clinical effectiveness to support patients through intensive treatment protocols have led to a virtual elimination of bone marrow as the source of cellular support for myeloablative chemotherapy in many transplant centers. Novel investigative approaches are also possible with PBPCs. In this review, the historical background of PBPCs is summarized, and the potential benefits (including economic advantages) of PBPCs to support dose-intensive chemotherapy for treating breast cancer are discussed. While dose intensification of breast cancer chemotherapy to the degree requiring PBPC support remains controversial and, in most centers, investigational, there is no doubt that PBPCs are an effective adjunct to the hematopoietic support of patients undergoing transplant-level cytotoxic treatments. Further study will undoubtedly lead to increased use of PBPCs in novel treatments for patients with breast cancer and other solid tumors. PMID- 8726589 TI - Peripheral blood progenitor cell transplantation for breast cancer: pharmacoeconomic considerations. AB - The recent observation that mobilized peripheral blood progenitor cells (PBPCs) can be used as a source of hematopoietic support has had a major effect on the cost and morbidity associated with bone marrow transplantation in patients with breast cancer. The pharmacoeconomic impact of this new technology is especially evident when secondary costs are analyzed. We see a reduction in hospital stay, decreased use of resources such as platelet transfusions and antibiotics, and long-term quality life-years gained for those patients benefiting from this therapy. Experienced transplant centers have found that the direct cost of high dose chemotherapy using filgrastim and PBPC support is reduced as much as 50% to the patient or their insurance company. Pharmacists will play a key role in optimizing the benefits of PBPC transplantation, particularly because this therapy is moving to the outpatient arena. This article will review the pharmacoeconomic impact of PBPC transplantation primarily in terms of secondary cost measures and quality of life and discuss the limited direct cost data available and the impact of this therapy on pharmacy practice. PMID- 8726590 TI - The future of cell therapy. AB - Peripheral blood has replaced bone marrow as a source of hematopoietic stem cells for autologous rescue after high-dose chemotherapy. Patients who receive peripheral blood stem cell (PBSC) transplants experience rapid and sustained hematopoietic reconstitution. As a result, transplant-related mortality is now less than 5% at many centers, and the cost of high-dose chemotherapy has decreased considerably. However, the relapse rate continues to be unacceptably high, and the collection of hematopoietic stem cells from peripheral blood is inconvenient, time consuming, and expensive. This article discusses the current status of novel technologies such as positive selection of hematopoietic stem cells, ex vivo expansion of hematopoietic progenitor cells, allogeneic PBSC transplants, and umbilical cord blood transplants. Several companies are actively developing devices that positively select hematopoietic stem cells. Because positive selection reduces the volume of infused cells, patients experience fewer adverse effects related to dimethylsulfoxide (DMSO) or lysed cells. These devices may also serve as an ex vivo method to remove ("purge") residual tumor cells. Positively selected hematopoietic stem cells may be expanded ex vivo to produce a large number of a specific population of hematopoietic cells. By adding cytokines that stimulate and activate lymphocytes, natural killer cells, and other immune effector cells, investigators could expand the number of immune effector cells with antitumor activity and then infuse them into patients as a form of adoptive immunotherapy. Finally, peripheral blood and umbilical cord blood are promising new sources of hematopoietic stem cells for allogeneic transplants. PMID- 8726591 TI - The risk of myocardial infarction associated with antihypertensive drug treatment in persons with uncomplicated essential hypertension. AB - We conducted a case-control study based on computer-recorded information accrued in the United Kingdom General Practice Research Database to assess and compare the relation between different antihypertensive drug therapies and myocardial infarction in patients with no known clinical or laboratory risk factors for myocardial infarction other than hypertension. Cases were treated hypertensive patients with no other known risk factors who developed a first acute myocardial infarction between January 1, 1993, and October 31, 1994. They were ascertained from a review of the clinical record together with a questionnaire filled out by the attending general practitioner. Controls were matched to each case for age, sex, general practice, and index date. Antihypertensive therapy was derived from the computerized patient record. The study consisted of 210 cases and 793 controls. Compared with users of beta-blockers alone, the adjusted relative risk (RR) estimates for all other treatment regimens were close to 1.0. A comparison of users of calcium channel blockers alone with users of beta-blockers alone yielded a RR estimate of 0.9 (95% CI 0.5, 1.7). We conclude that the risk of acute myocardial infarction in otherwise healthy, treated hypertensive patients is not materially associated with the particular drug they receive. PMID- 8726592 TI - Metformin in noninsulin-dependent diabetes mellitus. AB - Metformin is an oral antihyperglycemic agent that is approved by the Food and Drug Administration for the treatment of noninsulin-dependent diabetes mellitus. It differs from the sulfonylureas in that it is does not enhance insulin secretion and normally does not produce hypoglycemia. Metformin acts to decrease preprandial and postprandial blood glucose concentrations by increasing skeletal muscle uptake of glucose, decreasing gluconeogenesis, and decreasing absorption of glucose. The addition of metformin to maximum dosages of a sulfonylurea may synergistically improve glucose control. The drug may offer other potential benefits, such as weight loss or minimal weight gain, improved blood flow in patients with peripheral vascular disease, reduction of tissue plasminogen activator inhibitor, and improved lipid profiles. It is relatively safe if taken appropriately. Its most common side effects are gastrointestinal (nausea, diarrhea, anorexia), metallic taste, and vitamin B12 malabsorption. Lactic acidosis may also occur, but it is rare if metformin is avoided in patients with contraindications to its use. With careful monitoring, the agent may be considered for the initial treatment of obese patients who fail dietary measures, and those whose disease is refractory to maximum dosages of sulfonylureas or who do not tolerate them. PMID- 8726593 TI - Therapeutic options for treating major depression, and the role of venlafaxine. AB - Major depression is a debilitating disorder that is often undertreated. Psychotherapy, electroconvulsive therapy, and pharmacotherapy are options for management. Tricyclic antidepressants and selective serotonin reuptake inhibitors are the cornerstones of drug therapy. Venlafaxine, a phenylethylamine antidepressant that primarily inhibits reuptake of norepinephrine and serotonin, is an alternative to those agents. It has been studied in short-term and continuation studies and appears to have efficacy similar to that of imipramine, trazodone, and fluoxetine. Moreover, venlafaxine is effective in approximately one-third of patients with treatment-resistant depression. Venlafaxine is metabolized by the P-450 enzyme system to an active metabolite O-desmethyl venlafaxine, which is excreted renally. Nausea, somnolence, and dizziness are dose-related adverse effects that often occur with initiation of therapy. Increases in blood pressure, particularly with high dosages, also may occur. Drug drug interactions appear to be minimal. PMID- 8726594 TI - Hormone replacement therapy: estrogen after menopause. PMID- 8726595 TI - The mutated androgen receptor and its implications for the treatment of metastatic carcinoma of the prostate. AB - Androgen deprivation is the most effective therapy for patients with advanced prostatic carcinoma. The lack of androgen stimulation on these cells causes them to become apoptotic. Although therapeutic efficacy of initial androgen deprivation in prostate cancer is high, the emergence of androgen-independent cancer is inevitable. Withdrawal of the antiandrogen flutamide elicits surprising activity in these cancers. In numerous studies the response rates cell line harbors a mutation in codon 877 of the androgen receptor. The mutant receptor loses androgen specificity and is activated by various steroids as well as flutamide. Identical and similar mutations have now been isolated from human prostate cancer tissue. The discovery of the mutated androgen receptor sheds light on the emergence of androgen-independent cancer and should facilitate the development of more efficacious therapies. PMID- 8726596 TI - New pharmacologic approaches to acute spinal cord injury. AB - The incidence of spinal cord injury (SCI) in the United States is approximately 10,000 new cases per year. Strategies to prevent injury or salvage a few dermatomal levels may have significant effects on outcome. Several pharmacologic agents have been evaluated for their efficacy in patients with acute SCI. Methylprednisolone, when administered early, was the first drug to show significant improvement in outcome and is a standard of comparison for future agents. Several new drugs show promising results in animal models of SCI, with more extensive human trials currently under way. Results of more well-controlled clinical trials are necessary to determine which agents have significant neurologic benefits. PMID- 8726597 TI - The role of clarithromycin in the prophylaxis of disseminated Mycobacterium avium intracellulare infection in patients with AIDS. AB - Clarithromycin is a first-line agent in the treatment of disseminated disease due to Mycobacterium avium-intracellulare (dMAC) in patients with the acquired immunodeficiency syndrome, and its role in dMAC prophylaxis is nearing definition. Fifteen abstracts described clinical outcomes when clarithromycin was given in various dosages as monoprophylaxis or in combination with other agents. Patients totaled 1063, the majority of whom had CD4+ counts of 100 cells/mm3 or below. In one study, MAC disease occurred in 4.5% of 343 clarithromycin-treated patients compared with 12.6% of 341 receiving placebo (p < 0.001). The remaining studies combined to report only two positive blood cultures, one positive sputum culture, and one positive culture of unidentified origin during their respective study periods. Gastrointestinal intolerance caused discontinuation of therapy in 21 (2.9%) of 722 patients receiving clarithromycin. Clinical evaluations show that the drug decreases the risk of dMAC, prolongs survival, and is well tolerated. Controlled clinical trials continue. PMID- 8726598 TI - Drug-cytokine interactions: focus on cyclosporine. AB - The immune system is a complex network that regulates and maintains the host's defense system. Changes and alterations in the immune system precipitate a series of reactions to prevent further damage as well as initiate repair. The system's cellular component relies on cytokines (interleukins, tumor necrosis factor, interferons, etc.) to facilitate communication in response to a foreign antigen. Cytokine concentrations are therefore elevated during times of inflammation, such as rejection of a transplanted organ. Recent research suggests that interleukin-6 may have an inhibitory effect on cytochrome P-450 3A and thus affect drug metabolism. Cyclosporine, which is administered to prevent rejection of transplanted organs, is metabolized primarily by the P-450 3A system. Thus, the inhibitory effect of interleukin-6 may alter cyclosporine concentrations, which in turn may increase its adverse effects, such as nephrotoxicity. PMID- 8726599 TI - Resistance to nondepolarizing neuromuscular blocking agents. AB - Several case reports of resistance to short-term administration of nondepolarizing neuromuscular blocking agents (NNMBAs) have been reported in research and surgical settings. Recently, several reports documented resistance to NNMBAs during therapy for prolonged paralysis in critically ill patients. Adverse outcomes associated with NNMBA resistance may include inadequate ventilatory management or suppression of patient movement, and an increased risk of dose-dependent cardiovascular adverse effects. Pharmacoeconomic issues must be considered in that the cost of NNMBA therapy in a resistant patient may be significant. Although the specific etiologies of resistance are not clear, several pharmacodynamic and pharmacokinetic alterations may occur as a consequence of disease state or concomitant drug therapy. Pharmacodynamic changes include altered acetylcholine receptor physiology or sensitivity, inhibition of serum cholinesterase activity, and interaction with plasma constituents. Alterations in distribution volume, protein binding, and clearance may also contribute to resistance in several disease states. PMID- 8726600 TI - Combination therapy with low-dose lovastatin and niacin is as effective as higher dose lovastatin. AB - STUDY OBJECTIVES: To determine if low-dose lovastatin in combination with niacin causes a greater percentage reduction in low-density lipoprotein (LDL) cholesterol than lovastatin alone, and to determine if the combination increases the risk of serious adverse effects. design. Prospective, randomized, open-label, clinical trial. setting. Family medicine clinic of a university-affiliated hospital. Patients. Patients with fasting LDL cholesterol concentrations of at least 150 mg/dl after 4 weeks of dietary stabilization and washout of any cholesterol-lowering drugs. INTERVENTIONS: Twenty-eight patients received lovastatin 20 mg/day for 4 weeks after dietary stabilization and washout. If LDL cholesterol remained above 130 mg/dl (100 mg/dl in patients with coronary artery disease), they were randomized to receive either lovastatin 40 mg/day or a combination of lovastatin 20 mg/day and niacin 500 mg 3 times/day. MEASUREMENTS AND MAIN RESULTS: There was no difference in actual or percentage reductions of LDL cholesterol, total cholesterol, and triglycerides between the groups. A greater increase in high-density lipoprotein (HDL) cholesterol occurred with combination therapy (p = 0.024). There was no difference in liver function tests, glucose, or uric acid between the therapies. Based on drug-acquisition cost, combination therapy is approximately 40% less expensive than monotherapy. CONCLUSION: Low-dose niacin plus low-dose lovastatin was as effective as higher dose lovastatin in lowering total cholesterol, LDL cholesterol, and triglyceride levels. The combination may offer benefit in raising HDL cholesterol levels. PMID- 8726601 TI - Effect of food on the pharmacokinetics of a new hydroxypropyl-beta-cyclodextrin formulation of itraconazole. AB - STUDY OBJECTIVE: To compare the pharmacokinetics of a single 100-mg oral dose of itraconazole administered as 10 ml of a 10-mg/ml itraconazole solution in hydroxypropyl-beta-cyclodextrin under fasting versus postprandial conditions. DESIGN: Open-label, two-way, randomized, crossover study. SETTING: Janssen Research Foundation, Belgium. PATIENTS: Twelve healthy volunteers. INTERVENTIONS: Blood samples were obtained for pharmacokinetic analyses immediately before dosing and at regular intervals up to 96 hours after each dose. Blood and urine samples were obtained for hematologic, biochemical, and urinary safety analyses at baseline and at the end of the study. MEASUREMENTS AND MAIN RESULTS: The mean peak plasma concentrations of both itraconazole and its active metabolite hydroxy itraconazole were significantly higher under fasting conditions than under postprandial conditions. The mean times to peak concentration for both the parent compound and its metabolite were significantly shorter under fasting than under nonfasting conditions. The mean areas under the curve (AUC0-infinity and AUC0-24 hrs) were also significantly higher under fasting than under postprandial conditions. CONCLUSIONS: Our findings suggest that the higher bioavailability of this new formulation of itraconazole may be of benefit in seriously ill patients who are not able to ingest adequate quantities of food. The fact that the solution was also well tolerated and was not associated with clinically significant changes in any laboratory value further underscores the potential utility of this dosing form. PMID- 8726602 TI - The effect of cocaine on Ventricular fibrillation threshold in the normal canine heart. AB - We determined the effect of cocaine on ventricular vulnerability to fibrillation, as measured by ventricular fibrillation threshold (VFT), and cardiac electrophysiology in 20 anesthetized dogs with normal hearts. Animals were randomized in blinded fashion to receive a continuous 3-hour infusion of cocaine 0.11 mg/kg/minute (total dose 20 mg/kg) or placebo (lactose dissolved in normal saline). The VFT, systolic and diastolic blood pressures, ventricular effective refractory period (ERP), and electrocardiographic intervals were measured at baseline and every 30 minutes during infusion. Baseline mean +/- SE VFT in cocaine and placebo groups was 57.0 +/- 7.8 and 51.8 +/- 7.6 mA, respectively (p = 0.64). Cocaine did not significantly decrease VFT, but actually increased it (i.e., reduced ventricular vulnerability to fibrillation) compared with placebo (84.6 +/- 10.4 vs 55.8 +/- 7.2 mA, respectively, at 150 minutes, p = 0.04). Cocaine prolonged ERP and PR, QRS, QT, QTc, JT, and JTc intervals. Cocaine does not increase ventricular vulnerability to fibrillation in anesthetized dogs with normal intact hearts. Its electrophysiologic effects are similar to those of class I antiarrhythmic agents in this model. PMID- 8726603 TI - Electrophysiologic and electrocardiographic pharmacodynamics of cocaine. AB - To determine and describe relationships between plasma cocaine concentrations and electrophysiologic and electrocardiographic effects, 10 anesthetized dogs with normal intact hearts received a continuous 3-hour infusion of cocaine 0.11 mg/kg/minute (total dose 20 mg/kg). Data were collected as part of a randomized, blinded, placebo-controlled study investigating the effects of cocaine on ventricular fibrillation threshold. Every 30 minutes during infusion of cocaine or placebo and for 3 hours after discontinuation of the infusion, heart rate and mean arterial pressure were determined, effective refractory period (ERP) was measured, and QRS duration and PR, QTc, and JTc intervals were recorded. At the time of each 30-minute measurement, arterial blood was obtained to determine plasma cocaine concentrations. Hysteresis curves were observed for cocaine induced increases in ERP and PR interval. The effects of cocaine on QRS duration and QTc and JTc intervals were not well described by tested models. Pharmacodynamic modeling techniques may be used to describe relationships between plasma cocaine concentrations and specific cardiovascular effects of cocaine. Further study is required to determine applicability of this model for prediction of cocaine's cardiovascular effects in humans. PMID- 8726604 TI - The effects of sevoflurane and isoflurane on recovery from outpatient surgery. AB - This randomized, open-label study compared the investigational inhalational anesthetic sevoflurane with isoflurane in 47 healthy women undergoing elective ambulatory surgery. The women were randomized to receive either sevoflurane or isoflurane in 60% nitrous oxide-oxygen. Induction with thiopental 3-6 mg/kg was followed by vecuronium 0.1 mg/kg and fentanyl 0-200 micrograms. Duration of anesthesia, time to emergence, orientation, length of stay in the surgical unit, and hospital discharge were recorded. The emergence, length of stay, and discharge times after discontinuation of sevoflurane were 9.7 +/- 0.7, 120.6 +/- 8.0, and 244 +/- 15 minutes, respectively, and for isoflurane were 11.9 +/- 1.4, 106.8 +/- 7.1, and 282 +/- 24 minutes, respectively (NS). The isoflurane group had a higher frequency of postoperative cough. At the end of surgery, the sevoflurane group received a deeper level of anesthesia (minimum alveolar concentration 1.5 vs 1.3), however, these patients were oriented earlier (13.6 +/ 1.1 min vs 17.0 +/- 1.5 min isoflurane; p = 0.02) after discontinuation of anesthesia, although this difference is of little clinical significance. PMID- 8726605 TI - Oral administration of micronized progesterone: a review and more experience. AB - Historically, oral progesterone has been regarded clinically ineffective because of its poor absorption and rapid clearance. Recent evidence suggests that an oral micronized form of natural progesterone is readily absorbed, produces luteal phase serum concentrations, provokes an end-organ response, and has no detrimental effect on the lipoprotein profile. Thus it is considered by many to be an attractive alternative to synthetic progestin. We evaluated the effects of a single oral dose of micronized progesterone 300 mg in eight healthy postmenopausal women. The maximum serum concentration ranged from 15.72-625.98 ng/ml. The extent of absorption increased with increasing age. The reviewed literature and our data indicate considerable intersubject variability in the extent of progesterone absorbed after administration of oral micronized progesterone. PMID- 8726606 TI - Clearance of ticarcillin-clavulanic acid by continuous venovenous hemofiltration in three critically ill children, two with and one without concomitant extracorporeal membrane oxygenation. AB - Three children were receiving ticarcillin-clavulanic acid by continuous venovenous hemofiltration (CVVH). Two of them were also receiving concomitant extracorporeal membrane oxygenation (ECMO). We collected ultrafiltrate hourly to determine the clearance of ticarcillin-clavulanic acid by CVVH. Serum concentrations were also determined at the midpoint of each ultrafiltrate collection. All samples were collected over one dosing interval. The volume of distribution of ticarcillin and clavulanic acid was 0.26 +/- 0.01 and 0.69 +/- 0.23 L/kg, respectively. Total body clearance of ticarcillin, determined from the elimination rate constant and volume of distribution, was 0.038 +/- 0.003 L/kg/hour and for clavulanic acid was 0.18 +/- 0.03 L/kg/hour. The sieving coefficients for ticarcillin and clavulanic acid were 0.83 +/- 0.11 and 1.69 +/- 0.19, respectively. We attempted to estimate the clearances by ECMO, but the result was uninterpretable. PMID- 8726607 TI - Effective treatment of narcolepsy with codeine in a patient receiving hemodialysis. AB - A 64-year-old man with narcolepsy could not take stimulant drugs due to coronary heart disease. In the past he noted improvement in alertness when taking codeine for pain, but this was eventually discontinued. After he developed end-stage renal disease, and because the use of stimulants in this setting may be difficult, treatment with codeine was again initiated. This resulted in dramatic improvement in alertness and substantial reduction of cataplexy. Because it is simple to use and familiar to most physicians, codeine may be the drug of choice for narcoleptic patients who are undergoing hemodialysis. PMID- 8726608 TI - Does acyclovir increase serum lithium levels? AB - A 42-year-old woman was admitted to the hospital to receive intravenous acyclovir for a herpes zoster infection. At the time she was taking lithium carbonate 450 mg twice/day. Six days after starting acyclovir she exhibited signs of lithium toxicity. When measured, the serum lithium level had increased 4-fold during acyclovir therapy. Both agents are excreted by the kidneys, raising the possibility that acyclovir at high serum concentrations may interfere with the renal excretion of lithium. A MEDLINE search did not identify any citation describing the possibility of an interaction between the drugs. This case suggests that acyclovir when given intravenously in doses of 10 mg/kg may result in increased serum lithium concentrations. Until additional data are available, if intravenous acyclovir is administered concurrently with lithium, we recommend closely monitoring patients for signs of lithium toxicity and measuring serum lithium levels every second or third day. PMID- 8726609 TI - Thrombosis resulting from heparin therapy. AB - Heparin is widely used in current practice for a variety of indications. It is well known that it can cause thrombocytopenia, but not that thrombosis may also develop in thrombocytopenic patients and cause significant morbidity and mortality. A 56-year-old woman developed heparin-induced thrombocytopenia with thrombosis that resulted in the amputation of her leg. It is proposed that the reaction has an immune-mediated mechanism. Several ways of diagnosing the condition are available, specifically the serotonin-release assay and an enzyme linked immunosorbent assay. The investigational agent danaproid may be effective in the treatment of heparin-induced thrombocytopenia with thrombosis. PMID- 8726610 TI - Appropriateness of iron prescribing: a retrospective study. AB - We retrospectively reviewed the charts of 120 patients who received an initial prescription of iron from a resident physician in internal medicine to determine how accurately house officers diagnose iron-deficiency anemia before initiating iron therapy. Each patient's laboratory records were reviewed for the 3-month period before the prescription. Of the 120 patients, 77 (64%) did not have any iron tests performed to aid in the diagnosis of iron-deficiency anemia. Forty three percent of those who had iron tests did not have the disorder by our criteria. Ferritin levels and iron profiles are often not measured in patients prescribed iron, and when they are, they are frequently misinterpreted by medical house officers. This can lead to inappropriate gastrointestinal procedures as well as inappropriate prescribing of iron. PMID- 8726611 TI - Application of publication rates: another perspective. PMID- 8726612 TI - The development of clinical practice guidelines for treatment of peptic acid diseases in a VA ambulatory care clinic--a comment. PMID- 8726613 TI - A cost comparison of beta 2-agonist bronchodilators is not a cost-effectiveness comparison. PMID- 8726614 TI - Percutaneous hepatic venous isolation and extracorporeal charcoal hemoperfusion for high-dose intraarterial chemotherapy in patients with colorectal hepatic metastases. AB - The results of treating 12 consecutive patients with unresectable colorectal hepatic metastases with a hepatic arterial infusion of high-dose Adriamycin, 100 120 mg/m2, using hepatic venous isolation (HVI) and charcoal hemoperfusion (CHP) are reported herein. Adriamycin was administered over 5-15 min under extracorporeal drug elimination by HVI-CHP. HVI was percutaneously accomplished by either the double-balloon technique using a Fogarty occlusion catheter (8/22F) or a balloon-tipped catheter (16F). During the infusion, isolated hepatic venous blood was filtered by CHP and pumped into the left axillary vein. There were no lethal complications, and good hemodynamic tolerance to HVI-CHP was confirmed. Tumor liquefaction accompanied by a sharp decrease in serum carcinoembryonic antigen levels by more than 50% of pretreatment levels was observed in 6 of the 12 patients 1 month after treatment. Apart from chemical hepatitis, which developed in 11 (92%) of the patients, the Adriamycin toxicities were well controlled following the development of nausea and vomiting in 2 patients (17%), leukopenia < 2,000/mm3 in 3 (25%), and gastric ulcer in 1 (8%). These results indicate that this method is a safe and useful procedure for otherwise hazardous high-dose intra-arterial chemotherapy in patients with unresectable hepatic tumors. PMID- 8726615 TI - Preoperative administration of antibiotics in patients with suspected acute appendicitis. AB - The effectiveness of administering antibiotics preoperatively to patients with suspected appendicitis was evaluated over a 2-year period in 105 consecutive patients who presented with right lower quadrant (RLQ) tenderness and a white blood cell (WBC) count of over 10,000/mm3 or a temperature of over 37 degrees C. All the patients were preoperatively administered cefazolin (CEZ), cefotiam (CTM), or fosfomycin (FOM) except those with apparent peritonitis, which resulted in the recovery of 41 patients (39%). Of the remaining 64 patients, 14 (13%) had catarrhal appendicitis, 34 (32%), phlegmonous appendicitis, and 16 (15%), gangrenous appendicitis. Preoperatively, there were no significant differences among these groups in the WBC count or temperature. Following the administration of antibiotics, both these parameters decreased significantly in the patients with catarrhal appendicitis, although a slight RLQ tenderness persisted; in those with phlegmonous appendicitis, the WBC count decreased significantly, but the temperature remained elevated, with slight rebound tenderness in the RLQ; and in those with gangrenous appendicitis, there were no significant differences between the pre- and postoperative data in WBC count or temperature, and the abdominal symptoms did not change. These results show the value of administering antibiotics within 24h of the onset of symptoms indicative of acute appendicitis to allow time to evaluate the patients' condition, decide the operative indications, and prevent unnecessary laparotomy. PMID- 8726616 TI - The possible risk of lower-limb sclerotherapy causing an extended hypercoagulable state. AB - The risk of thrombosis after lower-extremity sclerotherapy is still an unresolved issue. This study was conducted to investigate the influence of sclerotherapy on coagulation and fibrinolysis by examining 20 patients who underwent surgical procedures, 10 of whom were treated by surgery alone (control group), while the other 10 were given sclerotherapy using 1% hydroxypolyaetoxydodecan as polidocanol (sclerotherapy group). Sex, age, and severity of disease was comparable between the two groups. No significant difference was found in the transient elevation of acute phase proteins, C-reactive protein (CRP), or fibrinogen. Thrombin antithrombin III complex (TAT), a marker of coagulation, transiently increased following treatment. In the control group, TAT peaked 3 days after treatment, whereas in the sclerotherapy group the elevation was prolonged, peaking 7 days after treatment. Elevation of the markers of fibrinolysis, plasmin plasmin inhibitor complex (PIC) and fibrin degradation products (FDP), was slower than that of TAT, peaking 7 days after treatment in both groups, the plasma PIC being significantly enhanced 7 days after treatment in the sclerotherapy group. A significant decrease in the platelet count was observed 3 days after treatment in the sclerotherapy group. These results suggest that sclerotherapy may enhance coagulation or fibrinolysis after surgical procedures. PMID- 8726618 TI - Pseudointimal hyperplasia of ridged outer wall polytetrafluoroethylene vascular prostheses. AB - In addition to the polytetrafluoroethylene (PTFE) vascular graft (G) with its conventionally smooth surface, a unique PTFE graft with a ridged outer wall (T) is now also currently available for clinical use. Although an excellent antikinking property is provided by this unique outer structure, the possible influence of the structure on the formation of pseudointima has not yet been investigated in detail. Four kinds of T grafts (3 mm inner diameter, 3 cm long) with various fibril lengths (FL, T-15, T-30, T-60, T-90) and a G graft with 30 microns FL (G-30) were implanted into the inferior vena cava of rabbits. The patency of the grafts at 4 weeks were as follows: 6/8(T-15), 6/8(T-30), 5/8(T 60), 0/8(T-90) and 4/6 (G-30). Pseudointimal hyperplasia (PH) of the T grafts advanced as the FL increased, judging by the thickness of the pseudointima, cellular density, and maturity of fibroblasts. In addition, the maturity of endothelial-like cells on the luminal surface increased as the FL increased. The degree of pseudointimal hyperplasia in G-30 was comparable to that of T-15, although the maturity of the endothelial-like cells was similar to that of T-60. Microscopically, there was a micro-heterogeneity of cellular density in T grafts probably due to the uneven outer structure. In conclusion, not only FL but also the outer structure of PTFE may thus influence the formation of the pseudointima. PMID- 8726617 TI - Continuous monitoring of short-latency somatosensory evoked potentials during cardiac and aortic surgery. AB - The effectiveness of monitoring somatosensory evoked potentials (SEPs) intraoperatively to detect brain damage early remains controversial. To assess the diagnostic accuracy of this modality, a study was conducted between 1991 and 1994, recording SEPs in 287 consecutive patients undergoing cardiac and aortic surgery using cardiopulmonary bypass (CPB) with moderate hypothermia or deep hypothermic circulatory arrest. From P1 to N2 of the SEPs occurring within 50 ms latency in response to electrical stimulation of the median nerve were recorded over the contralateral postcentral cortex at 5-min intervals using a Neuropack-2 (Nihon Koden, Tokyo, Japan). Normal SEPs were recovered in 247 patients postoperatively; however, 2 of these patients had suffered a cerebral infarction and 1, a transient stroke intraoperatively, demonstrating a false-negative incidence of 1.2%. On the other hand, three different types of abnormal SEPs were recorded postoperatively. P1 and N1 absence, probably caused by a subcortical lesion, was observed in 4 patients; P2 and N2 absence, probably caused by a cortical lesion, was observed in 8 patients; and a flat SEP, representing diffuse damage, was observed in 2 patients. Among these 14 patients with abnormal SEPs, 7 showed no neurologic disturbance at all, demonstrating a false-positive incidence of 50%. Thus, we concluded that when normal SEPs are recovered during weaning from CPB, the incidence of brain damage could be predicted at below 5%. Conversely, when abnormal SEPs are demonstrated, the incidence of brain dysfunction impeding a return to active life is estimated to be about 70%. PMID- 8726619 TI - Appendical fistulae formation as a complication of primary Crohn's disease prior to surgical management: report of a case. AB - We report herein the unusual case of a 55-year-old man who presented with primary Crohn's disease of the appendix with fistulae. In this patient there was no history of previous appendectomy. Although fistula formation around the orifice of the appendix after appendectomy for primary Crohn's disease of the appendix has been observed in a number of long-term follow-up studies, preoperative fistula formation is extremely rare. PMID- 8726620 TI - Psoas abscess complicating Crohn's disease: report of two cases. AB - We report herein the cases of two patients with Crohn's disease complicated by a psoas abscess. The first patient was a 29-year-old man who underwent definitive surgery after acute inflammation had been controlled by nutritional therapy. In the second patient, a 37-year-old man, the abscess required drainage under local anesthesia prior to surgery. Both patients have been free from recurrence of any abdominal symptoms for about 2 years since undergoing surgery. Although psoas abscess is still regarded as a rare complication, with the increasing prevalence of Crohn's disease in Japan it has become one of the most important complications requiring surgical intervention. PMID- 8726621 TI - Video-assisted thoracic surgery for thorascopic resection of giant bulla. AB - This report outlines our experience of 6 patients who underwent video-assisted thoracic surgery (VATS) using a linear endoscopic stapler to remove a giant bulla from the lung. Successful treatment with VATS was carried out in 4 patients, but the procedure needed to be changed to a thoracotomy in 2 patients - in one because of difficulty in single-lung ventilation, and in the other, due to a persistent air leak. Thus, we conclude that giant bulla without any associated severe respiratory failure can be an indication for VATS. PMID- 8726622 TI - Traumatic cervical tracheal disruption: report of two cases. AB - We report herein the cases of two patients who suffered tracheal disruption, both of whom underwent successful surgical treatment. The first patient was a 48-year old truck driver who suffered severe dyspnea after jamming his neck in a truck door. An endotracheal tube was unable to be inserted due to bleeding and thus, an emergency tracheostomy was performed. On admission massive subcutaneous emphysema was noted in the neck and anterior chest, and tracheal disruption was confirmed by a lateral neck X-ray, computed tomography (CT), and fiberscopy. An emergency end-to-end anastomosis of the trachea with insertion of a T-type silicon tube into the lower trachea was performed. The second patient was a 36-year-old man who suffered severe dyspnea after having his neck caught in a chain while driving a motorcycle. On admission, marked subcutaneous emphysema in the neck and paradoxical movement of the trachea were noted. Tracheal disruption was confirmed by a lateral neck X-ray and CT, and a similar operation to that of the first patient was performed. This type of injury is rare; however, lateral neck X-ray, CT, and fiberscopy proved extremely useful for making an accurate diagnosis following which successful emergency surgery was able to be performed, achieving good long-term results. PMID- 8726623 TI - Resected acinar cell carcinoma of the pancreas with tumor thrombus extending into the main portal vein: report of a case. AB - The incidence of acinar cell carcinoma has been reported to be about 1% of all pancreatic neoplasms, and pancreatic cancer combined with tumor growth extending into the portal vein is a rare condition. We herein report a case of acinar cell carcinoma of the pancreas with a tumor thrombus extending into the main portal trunk. Preoperative imaging of the portal vein, consisting of computed tomography (CT), magnetic resonance imaging (MRI), and angiography, revealed an oval shadow defect in the main portal trunk along with an irregular mass in the pancreatic head. At operation, we confirmed a tumor thrombus extending from a tumor in the pancreatic head into the main portal trunk via the pancreatoduodenal veins. A pancreatoduodenectomy combined with partial resection of the portal vein was thus performed under a temporary portal vein shunt from the ileocecal vein to the umbilical vein. Immunohistochemical examination for alpha 1-antichimotrypsin and electron microscopic examination confirmed the diagnosis of acinar cell carcinoma of the pancreas with a tumor thrombus in the portal vein. Surgical excision combined with portal vein resection may therefore improve the prognosis of selected patients with portal tumor thrombus. PMID- 8726624 TI - An analysis of breast cancer in Hungary: experience of the National Institute of Oncology, Budapest. AB - Breast cancer is the most common type of cancer in women throughout the world (18%), with a yearly morbidity that is already over half a million. Its incidence in Hungary is increasing in each age group, and the mortality is strikingly high. This study was conducted to assess the effectiveness of breast-conserving surgery in comparison to more radical surgery on a large group of patients. During the 15 years between 1980 and 1994, a total of 6,358 patients with primary breast cancer underwent surgery at the National Institute of Oncology in Budapest, Hungary, as breast-conserving surgery in 2,026 patients (31.8%). Breast-conserving surgery is being performed in an ever-increasing number of patients, followed by adjuvant radiochemotherapy in premenopausal patients, or radiohormone therapy in menopausal patients. During follow-up, recurrence was found in only 5.9% of the patients who underwent breast-conserving treatment. Thus, according to our experience, breast conservation therapy for the management of breast cancer offers favourable results, provided that the preconditions to this treatment are adequately met. PMID- 8726625 TI - Surgery for esophageal and cardia cancer in Hungary: a nationwide retrospective five-year survey. AB - A nationwide retrospective 5-year survey was conducted, examining the surgical treatment of cancer of the esophagus and the esophagogastric junction in Hungary. The population of Hungary is at low risk of developing esophageal cancer, with an associated mortality rate of 5.84 per 100,000 in 1992. During the 5 years between 1988 and 1992, a total of 1197 resections were performed for cancer of the esophagus and cardia, with 817 for esophageal cancer (in the cervical area in 40, the upper and midthoracic areas in 436, and the lower third in 341), and 380 for cancer of the cardia. Most of the procedures were total or subtotal esophagectomy, performed in 629 patients, with the stomach being used for replacement of the esophagus in 555 (88.2%) patients, the right colon in 46 (7.3%) and the left colon in 28 (4.5%). Transhiatal blunt esophagectomy was performed in 264 patients, representing 22.0% of all resections. The overall leakage rate was 21%, occurring in 251 patients, and the overall mortality rate was 13.5%, or 162 patients. A very strong correlation existed between the experience of the departments where surgery was performed and the mortality rate. PMID- 8726626 TI - Effectiveness of multivisceral resection in surgery for gastrointestinal cancers. AB - A retrospective analysis was conducted on 196 patients who underwent surgery involving multivisceral resection for adenocarcinoma of the abdominal digestive tract, 101 of whom were over 70 years old. Resection or removal of a neighboring organ was justified by either tumorous involvement of the organ, oncological principles, or the surgical technique required. Thus, in addition to the primary tumorous organ, two other organs were resected or removed in 134 patients, and three or more other organs were resected or removed in 62 patients. The mortality rate was 5% in the former group, and 16% in the latter group, showing an overall mortality rate of 7.5% for the 196 multivisceral operations. Moreover, the 5-year survival rate of patients with microscopic evidence of tumorous involvement of the resected neighbouring organs was significantly lower than that of those without any evidence of involvement. PMID- 8726627 TI - Comparison of the functional results of ileorectostomy and ileal pouch-anal anastomosis following total colectomy. AB - To evaluate the functional results of continence reconstruction techniques following total colectomy, an analysis of long-term follow-up, quality of life parameters, and recurrence of basal disease was conducted on 17 patients who underwent ileal pouch-anal anastomosis, and 16 who underwent ileorectostomy. Satisfactory results were achieved in 29 of the 33 patients, partial results were achieved in 3, and only 1 remained totally incontinent. While this outcome was independent of the reconstruction technique, a lower mean daily frequency of defecation, better day-night distribution, faster definitive recovery from surgery, a higher degree of rehabilitation, and a lower frequency of unplanned outpatient visits, hospitalizations, and minor complications were achieved after ileorectostomy than after pouch-anal anastomosis. Furthermore, better results were observed in patients who had undergone surgery for familial polyposis or complicated diffuse colonic diverticulosis, compared to those who had undergone surgery for inflammatory bowel diseases (IBD). During the postcolectomy stage, tumor recurrence was found in 5 of 7 patients with familial polyposis and histologically confirmed malignancy, compared to only 1 of 6 patients with negative histological results. Extracolonic manifestations developed in two patients with ulcerative colitis, and a small bowel obstruction occurred in one with Crohn's disease. These findings emphasize the importance of carefully selecting patients for each type of continence reconstruction method after total colectomy according to their individual requirements. PMID- 8726628 TI - An overview of vascular surgery in Hungary. AB - The practice of vascular surgery in Hungary was initiated in 1952 and its progress can be seen as beginning with a "trial" stage from 1952 to 1960. During the period from 1960 to 1980, the first of the operations known today were performed, and since 1991, vascular surgery has become routinely practiced, with the number of vascular reconstructions each year exceeding 6,000. From a population of 10 million inhabitants in Hungary, a total of 6,892 arterial reconstructions were carried out during 1990 by 150 vascular surgeons, with a total of 400 allocated beds. PMID- 8726629 TI - The antiproliferative effect of vitamin D3 analogues. PMID- 8726630 TI - The lanolin paradox. AB - Several puzzling aspects of the use of lanolin are discussed as "lanolin paradoxes', in analogy with the 'paraben paradoxes'. Lanolin in topical therapeutic agents sensitizes a high proportion of patients, whereas the same lanolin is 'safe' in cosmetics so widely used by millions of individuals. Patients with an allergic contact dermatitis to lanolin in a medication applied to a stasis ulcer can nevertheless use lanolin-containing cosmetics and not experience a reaction. Lanolin-sensitive individuals often show false-negative patch test reactions to unaltered lanolin. Patch testing with 30% wool wax alcohols used in the standard patch test tray cannot be considered a reliable method for detecting and confirming lanolin allergies. There are too many false positive and false-negative results using the standard patch test tray. PMID- 8726631 TI - Heterogeneous reactivity with LH7.2 and the first prenatal diagnosis of generalized recessive dystrophic epidermolysis bullosa among Japanese patients. AB - BACKGROUND: The heterogeneity of abnormal patterns of expression of type VII collagen in the skin of Japanese patients with generalized recessive dystrophic epidermolysis bullosa (g-RDEB) remains unclear, and the prenatal diagnosis of this condition has not yet been performed in Asia. OBJECTIVE: The present study was performed to clarify patterns of abnormal expression of type VII collagen among Japanese patients with g-RDEB, and to evaluate the first application of prenatal diagnosis for this condition in an Asian country. METHODS AND RESULTS: Only 2 of 8 Japanese patients with g-RDEB evaluated demonstrated a complete absence of type VII collagen at the skin basement membrane zone when tested with an LH7.2 monoclonal antibody. The other 6 patients revealed present, although diminished, LH7.2 reactivity. The mother of 1 patient who lacked reactivity to the LH7.2 monoclonal antibody sought prenatal diagnosis. Electron microscopy of fetal skin specimens obtained at 19 weeks' gestation showed mature anchoring fibrils with no separation of the dermis and epidermis. Indirect immunofluorescence revealed normal expression of type VII collagen. The fetus was diagnosed as being unaffected, and a normal female infant was delivered at 38 weeks' gestation. CONCLUSION: Our findings indicate a lower incidence of the negative expression of LH7.2 epitope in the type VII collagen among Japanese rather than non-Japanese patients with g-RDEB. However, LH7.2 still serves as a reliable diagnostic probe under certain conditions such as the diagnosis and prenatal diagnosis of g-RDEB as in the present case. PMID- 8726632 TI - Epidermal fatty-acid-binding protein in psoriasis, basal and squamous cell carcinomas: an immunohistological study. AB - BACKGROUND: In human keratinocytes, we have recently characterized a low molecular-weight cytosolic protein of 15 kD that specifically binds fatty acids (FAs) with high affinity, the epidermal FA-binding protein (E-FABP). The distribution of E-FABP in skin diseases is not known. OBJECTIVE: To localize by immunohistochemistry the expression of E-FABP in psoriasis, basal and squamous cell carcinomas in order to obtain indirect information, at the cellular level, on the transport of the FAs. RESULTS: E-FABP was localized in the upper stratum spinosum and stratum granulosum in normal and non-lesional psoriatic skin. In contrast, lesional psoriatic epidermis strongly expressed E-FABP in all suprabasal layers, like nonkeratinized oral mucosa. The basal layer did not express E-FABP reactivity in any of these samples. Accordingly, basal cell carcinomas were E-FABP negative whereas only well-differentiated cells of squamous cell carcinomas expressed E-FABP. CONCLUSION: It is unlikely that E-FABP plays a significant role in FA uptake by basal cells. Our data rather indicate that E-FABP expression is related to the commitment of keratinocyte differentiation and that the putative role of E-FABP should not be restricted to the formation of the skin lipid barrier. Since the pattern of E-FABP expression mimics cellular FA transport, our results suggest that lesional psoriatic skin and oral mucosa have a higher metabolism/transport for FAs than normal and non lesional psoriatic epidermis. PMID- 8726633 TI - Effects of systemic treatment with statins on skin barrier function and stratum corneum water-holding capacity. AB - BACKGROUND: Topical application of inhibitors of HMGCoA reductase, the rate limiting enzyme of cholesterol synthesis, has been shown to induce impairment of barrier function. OBJECTIVE: Assessing whether oral administration of statins used for reducing blood levels of cholesterol induces functional changes in stratum corneum barrier. MATERIALS AND METHODS: 69 subjects of both sexes under going treatment for hypercholesterolemia (mean age 48 +/- 11 years) entered the study; 43 had been treated with simvastatin and 11 with pravastatin for 6 months; 15 only on dietary regimen served as controls. Efficiency of stratum corneum water barrier was evaluated by transepidermal water loss (TEWL) measurement using an evaporimeter; water-holding capacity of the stratum corneum was assessed by the sorption-desorption test measured by capacitance. Statistical analysis was performed using ANOVA. RESULTS: No differences were found between the groups (simvastatin, pravastatin, diet) concerning both basal TEWL and the dynamic of water binding in the stratum corneum. CONCLUSIONS: Prolonged treatment with cholesterol-lowering drugs based on inhibition of HMGCoA reductase does not alter the permeability barrier of the skin. PMID- 8726634 TI - HPV-11- and HPV-16-associated oral verrucous carcinoma. AB - BACKGROUND: Verrucous carcinoma is a slowly growing, well-differentiated neoplasm with a predilection for the oral cavity. OBJECTIVE: Data on the association of human papillomavirus (HPV) with oral verrucous carcinoma are very scarce. We searched for the presence of HPV in different stages of the tumour in a patient repeatedly treated by means of surgery, cryotherapy, radiotherapy and chemotherapy over a decade. METHODS: HPV DNA was detected in archival tumour tissue by nested PCR. HPV types were identified based on restriction fragment analysis. RESULTS: HPV type 11 was found in 1 early-stage and HPV 16 in 2 late stage biopsy specimens of the tumour. CONCLUSION: Our findings provide further evidence for a role of HPV in oral verrucous carcinoma. The case history emphasizes the importance of a joint clinicopathological approach to this tumour. PMID- 8726635 TI - Clinical and pathophysiological aspects of hydroxyethyl starch-induced pruritus: evaluation of 96 cases. AB - BACKGROUND: It has been recently recognized that long-term infusions of hydroxyethyl starch (HES) as a substitute of human plasma may lead to deposits within the human skin and clinically induces severe pruritus in approximately one third of the patients treated. OBJECTIVE: To analyze the clinical features and pathophysiology of HES-induced itching. METHODS: Clinical data of 86 patients were analyzed retrospectively. Furthermore, 10 patients were followed prospectively, in whom we also performed routine laboratory tests, skin biopsies, allergological investigations (skin prick, basophil degranulation test), and substance P measurements in stimulated macrophages. RESULTS: Special features of HES-induced pruritus included long latency of onset and persistence. Noteworthy, itching is not always generalized. 30% of patients present with localized pruritus. Symptoms were not relieved by antihistaminics. Accordingly, the basophil degranulation test after HES exposure was negative. Also, substance P release from macrophages was not increased following HES stimulation. Deposits were found mainly in macrophages and endothelial cells. CONCLUSIONS: Our findings indicate that pruritus in HES-infused patients is most likely not triggered by pruritogenic mediators. We support data from the recent literature, suggesting a direct stimulation of cutaneous nerves by HES deposits. In patients suffering from long-standing itching it seems important to consider previous HES exposure as a possible cause not only in generalized pruritus but also in localized symptoms. PMID- 8726636 TI - Atopic eczema and histamine-induced sensations. AB - BACKGROUND: In previous studies we observed diminished pruritogenic effects and reduced flare reactions after iontophoretic application of histamine in patients with acute atopic eczema (AE). OBJECTIVE: To determine a possible mechanism acting on peripheral and central nervous components of the histamine-mediated itch reaction. METHODS: Forty-eight individuals with different predispositions to AE were included. According to an 'atopic score', 16 individuals had no history of atopic symptoms, another group of 16 individuals showed a disposition to AE and a further group of 16 individuals had a clear history of AE. However, all individuals were devoid of acute atopic symptoms and their IgE levels were below 150 U/ml. Histamine was iontophoretically applied to the skin of the forearm, and stimulus-induced itching and burning sensations were rated on visual analogue scales. RESULTS: Individuals with previous episodes of AE reported weaker itch sensations after histamine. In addition, the sensations were often described as burning by this group. CONCLUSION: The results suggest an altered central nervous responsiveness already in symptom-free individuals, since peripheral vascular reactions to histamine were not significantly reduced. PMID- 8726637 TI - The effect of 1,24(R)(OH)2D3 cream and ointment on epidermal proliferation and differentiation in mice. AB - BACKGROUND: The 1,24(R)(OH)2D3 (tacalcitol) ointment (2 micrograms/g) is available commercially as an antipsoriatic drug in Japan, but the cream preparation of tacalcitol is still under development. OBJECTIVE: This study was conducted to compare the ability of tacalcitol cream and ointment to inhibit epidermal proliferation and induce epidermal differentiation. METHODS: We measured the ornithine decarboxylase activity and type I transglutaminase activity as indices of proliferation and differentiation, respectively, in hairless mice. RESULTS: These effects were statistically equal to those of the ointment preparation at the same dose, 2 micrograms/g, without inducing hypercalcemia. CONCLUSIONS: These findings suggest that topical application of 1,24(OH)2D3 cream (2 micrograms/g) might have the same potency as the ointment formulation in the treatment of psoriasis. PMID- 8726638 TI - Interferon alfa-2a in the treatment of Behcet's disease. AB - BACKGROUND: There is no definite treatment for Behcet's disease. New drugs are being evaluated in cases unresponsive to conventional treatment modalities. OBJECTIVE: In this study we investigated the efficacy of interferon alfa-2a on skin lesions in 18 Behcet's disease patients who had predominantly mucocutaneous involvement. METHODS: Eighteen patients with Behcet's disease were treated with interferon alfa-2a at 3 million IU/day in the first week (three times a week), 6 million IU/day in the second week (three times a week), 9 million IU/day in the third week and thereafter (three times a week). Interferon alfa-2a was administered subcutaneously for a total of 12 weeks. RESULTS: At the end of the treatment the efficacy of the drug was found to be good in 9 patients and very good in 7 patients. Interferon alfa-2a is particularly effective in skin manifestations such as genital ulceration and erythema-nodosum-like lesions. It was also found to be effective in systemic manifestations such as fever, diarrhea and eye involvement. CONCLUSION: Our results indicate that interferon alfa-2a is a promising drug in the treatment of Behcet's disease. PMID- 8726639 TI - Emollients and photo(chemo)therapy: a call for caution. AB - BACKGROUND: Emollients used for skin care, treatment of a disease or lubrication, before irradiation during the course of phototherapy or photochemotherapy, may significantly interfere with the transmission of UV light and may influence the outcome of the therapy. OBJECTIVE: These experiments should characterize and quantify the possible interactions of usual emollients with UV light transmission. METHOD: The method was an in vitro technique similar to the one already used for the determination of sun protection factors of topical sunscreen preparations. RESULTS: Among the products tested, two showed a significant photoprotection capable of interfering with the course of therapy. Oils enhanced UV light transmission significantly. CONCLUSIONS: Emollients may show significant interference with UV light transmission during phototherapy or photochemotherapy. The outcome and course of the therapy could thus be significantly modified. PMID- 8726640 TI - Photodynamic therapy with topical application of 5-aminolevulinic acid in the treatment of actinic keratoses: an initial clinical study. AB - BACKGROUND: The incidence of actinic keratoses (AK) is rising and there is still a need for therapeutic alternatives. OBJECTIVE: To demonstrate the efficacy and tolerability of topical photodynamic therapy (PDT) using 5-aminolevulinic acid (ALA) in the treatment of AK. METHODS: Ten patients with 36 lesions (19 at hands and arms, 17 on the head) received ALA-PDT once (occlusive application of a 10% ALA emulsion for 6 h, irradiation with red light, 580-740 nm, 150 J/cm2) and were then monitored for 3 months. Therapeutic efficacy was judged using a score evaluating infiltration and keratosis of AK. RESULTS: After 28 days, significantly lower score sums were observed (head: mean = 15%; hand: mean = 67%) compared to the initial score (100%). Complete remission was achieved in 71% of AK localized on the head. No notable side effects were observed. CONCLUSION: This study demonstrated the potential of good efficacy and tolerability in the treatment of AK using topical ALA-PDT. How efficacy for lesions on the hand can be improved and whether PDT is able to concur with established treatment modalities remains to be shown in further studies. PMID- 8726641 TI - Topical treatment with 1% sodium cromoglycate in pyoderma gangrenosum. AB - BACKGROUND: Pyoderma gangrenosum is an uncommon skin disease usually treated with systemic drugs, most frequently corticosteroids. Isolated topical treatment is usually considered unsatisfactory. OBJECTIVE: To evaluate the effect of topical 1% sodium cromoglycate solution on pyoderma gangrenosum in 5 patients 25-30 years of age. METHODS: 5 patients, including 2 under systemic steroid treatment, hospitalized in our department from 1992 to 1993 because of pyoderma gangrenosum, were treated with topical 1% sodium cromoglycate solution. Systemic corticosteroids were further added in 2 patients whose initial improvement was inadequate. RESULTS: Initial improvement was noted in all 5 patients after 3-7 days of sodium cromoglycate treatment. Complete healing of the ulcers occurred within 5-8 weeks. CONCLUSIONS: Topical treatment with sodium cromoglycate can be effective as adjunctive or sole treatment in pyoderma gangrenosum. PMID- 8726642 TI - The hyperkeratotic variant of porokeratosis Mibelli is a distinct entity: clinical and ultrastructural evidence. AB - An isolated case with the hyperkeratotic variant of porokeratosis Mibelli (PM) is reported. The 62-year-old woman developed multiple brown macules with well demarcated large keratotic nodules on both legs. Histopathological studies revealed the presence of a cornoid lamella characteristic of porokeratosis. Ultrastructural investigations revealed dyskeratotic as well as edematous, swollen keratinocytes; autophagocytotic vacuoles were visible. The clinical and ultrastructural peculiarities of the hyperkeratotic variant of PM suggest the possibility of a separate entity. PMID- 8726643 TI - Cutaneous fibrous dysplasia: an incomplete form of the McCune-Albright syndrome. AB - A number of genetic disorders have been described in limited form. We report a patient with precocious puberty, large irregularly shaped cafe-au-lait spots since birth and a diffuse scalp alopecia showing the cutaneous changes of fibrous dysplasia. Histologically, the hair follicles were replaced by convolutions of fibrous tissue. This is to our knowledge the first patient reported with an apparently localized cutaneous form of this syndrome, a pattern predicted by the recently described somatic mutation of the Gs alpha gene. This is the second patient reported to our knowledge with the cutaneous McCune-Albright syndrome and scalp alopecia, and the first with diffuse scalp alopecia, the latter being the presenting sign. We believe that the differential diagnosis of both localized and diffuse alopecia should include the McCune-Albright syndrome. PMID- 8726644 TI - Coexistence of multiple perifollicular fibromas and colonic polyp and cancer. AB - Perifollicular fibroma (PFF) is a rare cutaneous hamartoma that shows differentiation to the connective tissue sheath of the hair follicle. It may be solitary or multiple; the latter has been suggested to exhibit autosomal dominant inheritance and association with colonic polyposis. We report a 72-year-old Japanese female with multiple PFF on the face, possibly familial, who developed colon cancer in addition to colonic polyps. PMID- 8726645 TI - A case of acrogeria associated with late-onset focal dermal elastosis. AB - We report a case of acrogeria. Biochemical analysis of patient fibroblasts demonstrated unaltered type III collagen synthesis. An RNA hybridization assay demonstrated that the levels of type I and elastin mRNAs were normal. The patient had multiple yellow papules on the neck, axilla and inguinal area which clinically and histologically resembled late-onset focal dermal elastosis. Both diseases may have some similar abnormal metabolism of connective tissue. PMID- 8726646 TI - Self-healing cutaneous mucinosis. AB - Self-healing juvenile cutaneous mucinosis is a new entity distinct from previously described forms of cutaneous mucinosis. We present a new case in a 26 year-old female patient who began abruptly with a papular eruption, involving the scalp, face, neck and trunk, accompanied by periarticular papules on the hands and arthralgias. No associated disease could be detected and the lesions resolved spontaneously within a few months. This is the first case described in an adult patient. PMID- 8726647 TI - Systemic scleroderma and perforating granuloma annulare: differential diagnosis from calcinosis. AB - BACKGROUND: Systemic scleroderma is a disorder of unknown etiology with skin sclerosis. Its major histological features are swollen and homogenized collagen bundles. OBJECTIVE AND METHODS: We describe 2 patients with systemic sclerosis who have multiple umbilicated nodules indistinguishable from calcinosis cutis. RESULTS: Histological examinations including Von Kossa staining revealed features of perforating granuloma annulare, but not of calcinosis cutis. CONCLUSION: The association may not be fortuitous but both diseases may be etiologically related. PMID- 8726648 TI - Nodular balloon cell component in a cutaneous melanoma of the superficial spreading type. AB - The rare histopathologic feature of a nodular balloon cell component in a malignant melanoma of the superficial spreading type in a 46-year-old male patient is presented. Immunohistochemical analyses proved the tumor cells to express the melanocytic markers S100 protein, NK1/C3 and HMB 45 antigens in an intense and distinct pattern. The controversial theories of the pathogenesis of balloon cell changes in melanocytic neoplasms are discussed. PMID- 8726649 TI - Febrile ulceronecrotic Mucha-Habermann disease: a case report and review of the literature. AB - A 32-year-old male with febrile ulceronecrotic Mucha-Habermann disease (FUMHD) responsive to methotrexate is reported. This is a severe variant of pityriasis lichenoides et varioliformis acuta characterized by the acute onset of a widespread ulceronecrotic cutaneous eruption together with high fever and systemic involvement. To our knowledge, only 13 patients with FUMHD have been reported to date. PMID- 8726650 TI - Dermatofibrosarcoma protuberans of the sole. AB - A 42-year-old Chinese man presented with a dome-shaped, red, tender, bony-hard tumor on his right sole. Histological examination revealed tumor nests which consisted mainly of fibroblast-like cells with large, spindle-shaped nuclei and histiocyte-like cells extending from the upper dermis to the subcutaneous tissue. The tumor cells showed both a storiform and a cartwheel pattern. The location of a dermatofibrosarcoma protuberans on the plantar aspect of the foot is exceptional. PMID- 8726651 TI - Vitiligo and Graves' disease following treatment of malignant melanoma with recombinant human interleukin 4. AB - This report describes a patient with metastatic malignant melanoma treated with recombinant human interleukin 4 who developed vitiligo and severe Graves' Disease after therapy. The patient has experienced complete remission of melanoma for 49+ months. The association of antitumor response, vitiligo and thyroid disorders is reviewed. PMID- 8726652 TI - Incontinentia pigmenti and Behcet's disease: a case of impaired neutrophil chemotaxis. AB - We describe a patient with incontinentia pigmenti who developed the clinical picture of intestinal Behcet's disease. Diagnosis of Behcet's disease was base on a recurrent genital and oral ulcer and a positive HLA-B51. Impaired neutrophil chemotactic activity and an elevated plasma IL-6 level were found. Chromosomal study revealed no abnormality in the patient or her family. We discuss the possibility that there are common immunological abnormalities in the two syndromes. PMID- 8726653 TI - Cutaneous T-cell lymphoma associated with granulomatous slack skin. AB - Granulomatous slack skin disease (GSS) is a rare disorder characterized by bulky cutaneous lesions and epithelioid and giant cell granulomas with destruction of the dermal elastic tissue. We detail the observation of a 29-year-old man with clinical and histological features of GSS. Pendulous skin tumors were associated with typical clinical and immunohistochemical aspects of mycosis fungoides and with clonal rearrangement of the V gamma T-cell receptor gene in lesional skin. This case report supports cutaneous T-cell lymphoma as a cause of GSS. PMID- 8726654 TI - Septal panniculitis associated with infectious mononucleosis. PMID- 8726655 TI - Cilium circinatum (circle lash, ringelwimper, cil enroule) PMID- 8726656 TI - Trichodynia. PMID- 8726657 TI - Familial background in aberrant mammary tissue is a 'protective' factor against the development of nephrourinary anomalies. PMID- 8726658 TI - A Chinese cream (fu suo) for psoriasis. PMID- 8726659 TI - Pustular drug eruption induced by ferrous fumarate. PMID- 8726660 TI - CCAAT/enhancer binding protein is involved in the expression of the tumour necrosis factor gene in human monocytes. AB - Within the human TNF promoter we have identified two sites at positions -189 and 101 that show C/EBP specific binding of nuclear proteins from cells of the human monocytic line Mono Mac 6. Supershift analysis with anti C/EBP antibodies revealed that the complexes formed consist of both C/EBP alpha and C/EBP beta. When studying reporter constructs with a 5'-deletion series of the TNF promoter in cotransfection experiments with a C/EBP beta expression plasmid, a construct with the -1064 TNF fragment gave 26-fold transactivation, the -630 fragment showed 23-fold transactivation and the -107 fragment (containing the -101 C/EBP binding motif) still gave 16-fold transactivation. Mutagenesis of the -101 site in the -630 construct resulted in a reduction of C/EBP driven transactivation from 26-fold to 7-fold. Finally, when Mono Mac 6 cells were transfected with these constructs, stimulation by LPS induced a 19-fold transactivation in the 630 wild type construct, while the -630 construct carrying the -101 mutation was transactivated only 4-fold. Hence, the data indicate that the -101 C/EBP motif is crucial for TNF gene expression in human monocytes. PMID- 8726661 TI - 1 alpha, 25-dihydroxyvitamin D3 (calcitriol) induces apoptosis in stimulated T cells through an IL-2 dependent mechanism. AB - The steroid hormone 1 alpha,25-dihydroxyvitamin D3 (calcitriol) has been shown to inhibit PHA-induced T-cell proliferation. In the present work we show that calcitriol caused a concentration-dependent decrease in the viability of PHA stimulated but not unstimulated T lymphocytes. Under microscopy, nonviable cells exhibit classic features of programmed cell death or apoptosis. In addition, the same concentration of calcitriol induces a blockage in the G1 phase of T cell cycle and decreased secretion of IL-2, which was correlated with the extent of apoptosis. Thus, addition of exogenous IL-2 reverted both the cell cycle block and the apoptotic cell death induced by calcitriol. This data indicates that calcitriol induced apoptosis in PHA-stimulated T cell, and suggests that this action is mediated through suppression of IL-2 secretion. PMID- 8726662 TI - Signal transduction in Th clones: target of differential modulation by PGE2 may reside downstream of the PKC-dependent pathway. AB - Lymphokines produced by non-transformed Th clones, Th1 and Th2, were classified into three groups based on their patterns of expression by different stimuli: Group I, GM-CSF and IL-2, characterized by a strict requirement of activation of both the PKC- and calcium-dependent pathways; Group II, IFN-gamma, IL-3, and IL 4, partially induced by calcium ionophore alone; and Group III, IL-5, IL-6, and IL-10, partially induced by either PMA or calcium ionophore alone. Transfection of constitutively active PKC or p21ras replaced the requirement for PMA in expression of these lymphokines, with the exception of GM-CSF. Production of Group II lymphokines was partially induced by constitutively active calcineurin. Production of Group I and II lymphokines was highly sensitive to cyclosporin A, while Group III lymphokines were relatively resistant. Addition of prostaglandin E2 (PGE2) and overexpression of catalytic subunit of protein kinase A inhibited lymphokine production in Th1 cells, but not in Th2 cells, with the exception of GM-CSF. Production of Group III lymphokines induced by PMA alone was upregulated by PGE2, but that of Group II and III lymphokines induced by calcium ionophore alone was not affected. These results suggest that one of the targets of PGE2 is downstream of the PKC-dependent pathway. PMID- 8726663 TI - Differential effect of IFN-alpha and IFN-gamma on phosphorylation of p65 and p50 (rel) in the K562 cell line: implications for altered interaction with RXR beta. AB - Phosphorylation of nuclear transcription factors plays an important role in their ability to regulate genes in a differential manner. Recent studies indicate that the rel homology domain mediates interaction of rel proteins with other transcription factors. The rel domain is multiply phosphorylated. Thus, altered phosphorylation in this domain would affect synergy of rel proteins with members of other transcription factor families. This could affect differential expression of genes that are regulated by a combination of the two transcription factors. We have observed that in the K562 cell line IFN-alpha treatment leads to deficient phosphorylation of nuclear p65/p50 (rel). We propose that this prevents interaction between p65/p50 and RXR beta in IFN-alpha treated K562 cells. This could be a major reason for the lack of globin gene transcription by IFN-alpha. PMID- 8726664 TI - Expression of an efficient small molecular weight tumour necrosis factor/lymphotoxin antagonist. AB - TNF-alpha and lymphotoxin are proinflammatory cytokines that are non-homologous in sequence, have similar homotrimeric structure and they exert their biological activity by aggregating two types of shared cell surface receptors. The natural inhibitors of TNF and lymphotoxin are the shed extracellular domains of the p55 and p75 TNF receptors. However recombinant inhibitors composed of the extracellular domains of p75 or p55 receptors dimerized on IgG backbone have been shown to be much more effective. We have produced a dimeric form of the human p75 TNF receptor extracellular domain based on the structure of the native soluble shed receptor. The dimer was engineered by genetically linking the monomeric forms with a polyglycine-serine linker. Biochemical characterization showed that this dimeric TNF receptor elutes from a TNF affinity column at a lower pH than the monomeric form. Biological assay revealed this novel antagonist to be as efficient as a dimer based on an immunoglobulin backbone. However this new dimer is smaller, stable, and could have greater penetration into tissues. PMID- 8726665 TI - Differential effects of endotoxin and cytokines on prostaglandin E2 formation in cerebral microvessels and brain parenchyma: implications for the pathogenesis of fever. AB - Prostaglandin(PG) E2 is regarded as an essential mediator in the central action of pyrogens and fever. However, it is not clear how the appearance of cytokines in the circulation leads to the rise of PGE2 in brain (fever to an external noxa), nor is it clear whether bacterial toxins originating within the brain activate PGE2 directly or via the cytokines (fever to a central noxa). We have previously reported that human interleukin 1 (hIL-1) has no effect on PGE2 synthesis in isolated, feline cerebral microvessels. Since cytokine action may be species-specific and interleukin 6 (IL-6) is considered as important as IL-1 for fever, we have now examined the response of isolated, murine cerebral microvessels to homologous and heterologous IL-1 beta (rIL-1 beta and hIL-1 beta), heterologous IL-6 (hIL-6), and endotoxin. The same pyrogens were tested on rat cerebrocortical minces. We have found that PGE2 formation in the microvessels is not changed by either IL-1 beta (both forms) or hIL-6. Conversely, in brain minces rIL-1 beta (but not hIL-1 beta or hIL-6) is a PGE2 activator. Endotoxin stimulated PGE2 synthesis in both preparations and its action in brain was fully reversed by the hIL-1 receptor antagonist (hIL-1ra). Our data indicate that the cerebral microvasculature does not lend itself to a transducing function in the fever to an external noxa. In addition, they point to a mediator role of IL-1 in the fever to a central noxa. PMID- 8726666 TI - Interleukin-1 beta converting enzyme inhibition blocks progression of type II collagen-induced arthritis in mice. AB - To IL-1 beta is a principal mediator in the pathogenesis of inflammatory disease. The IL-1 beta-converting enzyme (ICE), a novel cysteine protease, is required for processing of the 31 kDa IL-1 beta precursor to generate the 17 kDa proinflammatory mature form. We investigated the effect of two irreversible peptidyl ICE inhibitors, VE-13,045 and VE-16,084, on IL-1 production in vitro and in vivo in acute and chronic inflammatory disease models. In vitro, VE-13,045 and VE-16084 inhibited IL-1 beta secretion by LPS-stimulated human adherent mononuclear cells (IC50's of 0.4 microM and 2.0 microM, respectively) and murine splenic monocytes (IC50's of 10 microM and 1.3 microM, respectively). Both VE 13,045 and VE-16,084 also inhibited LPS stimulated IL-1 alpha secretion, although with reduced potency. In vivo, a single intraperitoneal dose of VE-13,045 (50 mg/kg) administered to mice 60 to 75 minutes after a 40 mg/kg LPS challenge significantly reduced IL-1 beta serum levels by 50 to 70%. In the DBA/1J mouse model of Type II collagen-induced arthritis, prophylactic treatment with VE 13,045 (50 and 100 mg/kg/day) significantly delayed the onset of inflammation, with a 60% overall reduction in disease severity. VE-13,045 was more effective than either indomethacin (2 mg/kg/day) or methyl prednisolone (10 mg/kg/day). VE 13,045 was also effective in reducing inflammation and progression of arthritis when administered to mice with established disease. Histological analysis of wrist joints showed a reduction in synovial membrane damage, inflammatory cell infiltration and fibrosis, and cartilage erosion in VE-13,045-treated animals. This is the first demonstration of efficacy for an ICE inhibitor in a chronic disease model and suggests that ICE is an important target for design of anti inflammatory or disease modifying drugs. PMID- 8726667 TI - Preventive effect of hepatocyte growth factor on acute side effects of cyclosporin A in mice. AB - Cyclosporin A (CsA) is a potent, widely-prescribed immunosuppressant which has serious side effects. When recombinant human hepatocyte growth factor (rh-HGF) was co-administrated with CsA to mice, the severe digestive and/or neurological symptoms and degenerative changes in renal tubular cells and hepatocytes seen with cases of CsA administration were remarkably attenuated and mortality linked to CsA-administration was prevented by rh-HGF. HGF-administration stimulated the DNA synthesis of hepatocytes and renal tubular cells and facilitated reconstruction of hepato-renal tissue structure in vivo. Induction of HGF mRNA expression in the liver and kidney in CsA-administered mice was suppressed in the early stage of organ injury, while HGF mRNA levels increased in the lung three days after CsA-treatment. These observations suggest that the biological action of endogenous HGF is partly impaired after CsA-induced organ injury. Importantly, HGF had no apparent effect on the CsA-induced suppression of interleukin-2 mRNA expression in vitro, thereby indicating that the immunosuppressive potential of CsA was not affected by HGF. Whether or not HGF will prove to have such positive effects in patients requiring CsA-treatment is the subject of ongoing study. PMID- 8726668 TI - Circulating serum levels of IL-1ra in patients with relapsing remitting multiple sclerosis are normal during remission phases but significantly increased either during exacerbations or in response to IFN-beta treatment. AB - Interleukin (IL)-1 receptor antagonist (IL-1ra) is a naturally occurring inhibitor of IL-1 which binds to IL-1 receptors without generating immunologic responses. Evidence has recently been provided that the balance between the production of IL-1 and IL-1ra might influence the course of immunoinflammatory diseases such as inflammatory bowel diseases, rheumatoid arthritis (RA) and Lyme arthritis. To assess whether endogenous IL-1ra may also have a role on the course of multiple sclerosis (MS) we presently studied the fluctuation of the serum levels of IL-1ra in patients with relapsing remitting (RR) MS either during remission or exacerbation. Moreover, to evaluate whether the beneficial effect of IFN-beta on the course of MS might also be mediated by an increased production of IL-1ra, we measured the levels of circulating IL-1ra in MS patients prior to and after 6 months of continuous treatment with natural human IFN-beta (6,000,000 IU three times a week for 6 months). Our results demonstrated that, relative to control subjects, IL-1ra serum levels are "normal' during remitting phases of RR MS but significantly elevated either during exacerbations or in response to IFN beta treatment. PMID- 8726669 TI - A soluble interleukin 6 receptor isolated from conditioned medium of human breast cancer cells is encoded by a differentially spliced mRNA. AB - The human interleukin 6 receptor (IL-6R) is expressed on cells as a transmembrane protein of 80 kDa (gp80, ligand binding unit), or as a smaller soluble counterpart (sIL-6R, approximately 55 kDa). Recombinant or natural sIL-6Rs bind IL-6 and stimulate biological activity by association with the signal transducing subunit gp130 at the cell surface. The origin of sIL-6Rs is not clear. Haematopoietic cells express, in addition to the gp80 mRNA, an IL-6R mRNA where the transmembrane domain is spliced out, predicting a shorter protein with a modified basic sequence at the C-terminus. We show that the spliced mRNA is expressed in human T47D breast carcinoma cells and soluble IL-6R protein is indeed secreted by these cells. An antibody against the C-terminus of the spliced protein detects a 55-65 kDa glycosylated species in sIL-6R purified from T47D supernatant by classical and immunoaffinity chromatography. The spliced T47D IL 6R, glycosylated or after removal of O- and N-linked polysaccharides, has the same size as a recombinant spliced IL-6R from CHO cells. The recombinant spliced IL-6R acts on cells as an IL-6 agonist to stimulate transcription from IL-6 inducible enhancers. PMID- 8726670 TI - Leukaemia inhibitory factor (LIF) production in pleural effusions: comparison with production of IL-4, IL-8, IL-10 and macrophage-colony stimulating factor (M CSF). AB - Leukaemia inhibitory factor (LIF), a pleiotropic cytokine detected in various inflammatory body fluids, plays a poorly defined role in the pathogenesis of human disease. This study was conducted to correlate the LIF concentrations in pleural effusions with the type of pathology and to compare its levels with those of IL-4, IL-8, IL-10 and M-CSF for a given pathology. Pleural fluids from 97 patients were assayed for cytokines by specific ELISAs. The concentrations of all cytokines tested were higher in infectious pleural effusions than in other pathologies (malignant or transudative). The lowest levels were observed for transudates. Significant differences were noted between pathology groups for each cytokine. A good correlation was observed between LIF and IL-8 for malignant effusions [regression correlation coefficient (RC) = 0.480, P < 0.01], between LIF and IL-4 for infectious disorders (RC = 0.543, P < 0.05) between LIF and IL 10 for transudates (RC = 0.798, P < 0.001) and between M-CSF and IL-8 in all pathologies tested except for primitive neoplasia (P < 0.05). The LIF concentration in pleural space seems to be strongly associated with the intensity of inflammatory reaction. The LIF production appears to have different regulatory patterns between aetiologic groups. PMID- 8726671 TI - Release of interleukin 6 and activation of complement during and after paediatric cardiopulmonary bypass. Effect of autotransfusion of shed mediastinal blood and ultrafiltration. AB - Interleukin-6 (IL-6) and the complement activation products C3bc and terminal complement complex (TCC) were measured in three groups of children undergoing open heart surgery. One group was treated with intraoperative extracorporeal ultrafiltration and postoperative autotransfusion of shed mediastinal blood, one group was subjected to autotransfusion only and in one group none of these procedures were performed. No differences between the groups were observed concerning the degree of complement activation. Peak and total accumulated level of IL-6 was significantly higher in the group subjected to ultrafiltration and autotransfusion compared to the group treated conventionally with no interventions. IL-6 may be a sensitive marker of maneuvres increasing the inflammatory load during and after open heart surgery in children. PMID- 8726672 TI - Effects of superoxide dismutase and catalase on growth of retinal pigment epithelial cells in vitro following addition of linoleic acid or linoleic acid hydroperoxide. AB - The rod outer segments of the retina that are phagocytized by retinal pigment epithelial (RPE) cells are susceptible to lipid peroxidation because of their high content of polyunsaturated fatty acids. Linoleic hydroperoxides (LHP), synthesized by peroxidation of linoleic acids (LA), produce greater damage to retinal function than does LA. We compared the effects of LHP and LA on the growth of cultured chick embryonic RPE cells and analyzed a model of data sets using multiple linear regression for the number of cells as a function of time. The spectrum of LA had a sharp peak at 205 nm and a broad spectrum at 235 nm, while LHP had only a broad spectrum at 235 nm. Exposure to LA and LHP caused dose dependent damage of chick embryonic RPE cells: they were significantly more affected by the addition of LHP than LA. The antioxidative enzymes catalase and superoxide dismutase minimized damage to the RPE cells caused by LHP in proportion to the enzyme concentration. However, RPE cells incubated with LA were more affected by the enzymes than when no enzymes were added. PMID- 8726673 TI - Elevation of cGMP with normal expression and activity of rod cGMP-PDE in photoreceptor degenerate labrador retrievers. AB - Cyclic guanosine 3',5'-monophosphate (cGMP) levels were determined in retinas from a strain of Labrador Retrievers with inherited retinal dystrophy manifesting at early stages of retinal differentiation. The cGMP contents of dystrophic retinas of dogs from 1 to 4 months of age (n = 7) were significantly higher (p = 0.001) than in age-matched controls of the same breed (n = 11). Ultrastructure along the vertical retinal meridian was studied in developing retinas and findings were related to those of age-matched wild-type controls of the same breed. Slow central to peripheral progression of degeneration was observed in affected dogs. No differences were found in total cGMP-phosphodiesterase (PDE) activity, in PDE subunit composition as determined by Western blotting of 2-month old homozygote affected retinas, or in the amino acid sequence deduced from the nucleotide sequence of the PDE beta-subunit as compared to controls. This model of photoreceptor degeneration thus is the first case of an apparent abnormality of cGMP metabolism that is not associated with a defect in the PDE catalytic subunits, and it is also the first reported model not associated with severe developmental abnormalities and rapid degeneration. PMID- 8726674 TI - Ultrastructural study of the cytoskeleton of optic nerve axons in guinea pigs as revealed by a quick-freezing, deep-etching method. AB - The ultrastructure of the cytoskeleton of optic nerve axons in guinea pigs was examined by the quick-freezing and deep-etching (QF-DE) method. The optic nerve tissues were treated with 0.5% saponin before QF. In the replicas, the axoplasm, as observed with conventional ultrathin sections, was seen to be composed of longitudinally oriented microtubules (MT) and neurofilaments (NF). Thin and elaborate cross-linking structures were observed in the interstices between MTs, NFs, and membranous organelles. They consisted of two different types: NF associated cross-linking structures (about 20-50 nm in length) and MT-associated structures (about 10-20 nm in length), and may play a role in the slow transport of NF and MT and fast transport of organelles, respectively. PMID- 8726675 TI - Sympathetic innervation of the rat choroid: an autoradiographic tracing and immunohistochemical study. AB - The sympathetic innervation of the choroid was investigated by means of the anterograde tracer 3H-leucine, injected into the rostral part of the superior cervical ganglion. The tracer was autoradiographically visualized at the light- and electron-microscopic levels. Labelled unmyelinated fibres were found in the choroid and labelled terminals were observed in close proximity to the smooth muscle cells of arterioles. The labelled terminals contain granular vesicles of different size and clear vesicles; the mitochondria were of the electron-dense type and no cell-to-cell synaptic contacts were observed. In addition, tyrosine hydroxylase immunocytochemistry was performed. Immunostained terminals found in the choroid show ultrastructural features similar to those found in the tracing experiments: granular vesicles of different size, clear vesicles and electron dense mitochondria. On the basis of previous studies and the present observations, it is concluded that the choroid has a dual innervation, a parasympathetic innervation from the pterygopalatine ganglion and a sympathetic innervation from the superior cervical ganglion. The dual innervation is most likely involved in maintaining the homeostasis of vasodilatation and vasoconstriction in the choroid. PMID- 8726676 TI - Phospholipids in meibomian gland secretion. AB - The bulk of the lipid layer overlying the aqueous portion of the precorneal tear film is composed of polar and nonpolar components. The nonpolar lipids have been the subject of numerous studies; however, the polar lipids have remained relatively uncharacterized. The polar lipids are thought to contain surfactant phospholipids that are critical to the spreading of a lipid film over the aqueous layer, by providing an interface between this layer and the nonpolar lipids. The purpose of the present study is to identify and quantitate the phospholipid complement of meibomian gland secretion which provides the tear film with phospholipids. Meibomian gland secretion was collected from rabbits and phospholipids identified and quantitated by 31P nuclear magnetic resonance spectroscopy. Ten phospholipids were detected from meibomian gland secretion: diphosphatidylglycerol, dihydrosphingomyelin, ethanolamine plasmalogen, phosphatidylethanolamine (PE), phosphatidylserine, sphingomyelin, lysophosphatidylcholine, phosphatidylinositol, alkylacylphosphatidylcholine, and phosphatidylcholine (PC). The two major phospholipids were PC and PE, together comprising nearly 60% of the total phospholipid profile. The nature and relative concentrations of the meibomian gland secretion phospholipids are congruous with a surfactant role at the aqueous-lipid interface and, considering the physical chemistry of the tear film, suggest that the phospholipids should be organized in a very flat or planar configuration. PMID- 8726677 TI - Ultrastructural, immunohistological and biochemical characterization of cultured mouse corneal epithelial cells. AB - We developed a method to culture mouse corneal epithelium. Cultured cells tested by 1-D-SDS-PAGE exhibited protein mobility patterns similar to freshly isolated epithelia. Western blots with antibodies broadly recognizing cytokeratins showed a similar pattern for both fresh and cultured cells, but only the fresh sample stained with J7, specific for a 55-kD 'corneal' cytokeratin. Cultured cells examined at confluency by transmission electron microscopy exhibited desmosomal contacts typical of epithelia. The ability to culture mouse corneal epithelial tissue will be useful for studies requiring large amounts of material by reducing animal numbers. PMID- 8726678 TI - The further metabolism of sorbitol-3-phosphate and fructose-3-phosphate in the mature rat lens. AB - Glucose and phosphorus metabolism in mature (8-month-old) rat lenses were examined with NMR spectroscopy. Nondiabetic mature lenses contained sorbitol-3 phosphate (S3P) and fructose-3-phosphate (F3P) which were absent from young (1- to 2-month-old) normal rat lenses. The concentrations of these two phosphates can be changed through (1) diabetes induction with streptozotocin - this results in a dramatic increase in both compounds; and (2) oral dosing with a drug known to prevent sorbitol production - both metabolites disappeared. When normal mature lenses were incubated in 35.5 mM 13C1-glucose, both 13C1-lactate and 13C3-lactate were produced. Preservation of the 13C label at C1 is likely through the formation of 13C1-S3P and -F3P, which were then split through an aldolase-like mechanism into two 3-carbon compounds, one an unlabeled glycerol and the other 13C1-alpha-glycerophosphate (from S3P) and 13C1-dihydroxyacetone phosphate (from F3P). These reactions can contribute to the increase in alpha-glycerophosphate observed in both the streptozotocin-induced diabetic lenses and lenses incubated in high glucose. PMID- 8726679 TI - Prevention of secondary cataract by antimitotic drugs: experimental study. AB - To prevent the development of secondary cataract, mitomycin C (MC, 0.2 mg/ml) and 5-fluorouracil (5-FU, 50 mg/ml) were introduced in the irrigating solution (BSS Plus) during an extracapsular lens extraction in albino rabbits. Cellular multiplication in the whole lens capsule was quantified by means of flow cytometry. The total number of residual cells in the capsule after lens extraction together with the mitotic phases of the cell cycle were studied. A statistically significant difference (p < 0.005, Student t test) was obtained between the two groups treated with antimitotics when compared with the control group regarding the inhibition of mitosis in the capsular cell population together with the total cell number. However, 5-FU has shown a more potent inhibitory effect than MC (p < .0.005, Student t test). We did not record any ocular complication during a postoperative period of 2 months. MC and 5-FU could be a promising alternative in the prevention of posterior capsule opacification. PMID- 8726680 TI - Characterization of a cis-regulatory element and trans-acting protein that regulates transcription of the angiotensin II type 1a receptor gene. PMID- 8726681 TI - Human AT1 receptor gene regulation. PMID- 8726682 TI - Regulation of gene transcription of angiotensin II receptor subtypes in the heart. PMID- 8726683 TI - Sodium induced regulation of angiotensin receptor 1A and 1B in rat kidney. PMID- 8726684 TI - Characterization and regulation of angiotensin II receptors in rat adipose tissue. Angiotensin receptors in adipose tissue. AB - Characterization and regulation of angiotensin II (AII) receptor binding sites was performed in rat membrane preparations from nonadipose (liver, lung) and adipose (interscapular (ISBAT) and periaortic (PA) brown adipose tissue; epididymal (EF) and retroperitoneal (RPF) white adipose tissue). In membrane preparations from brown and white adipose sources, [125I]AII saturation binding revealed a single, high affinity (Kd range of 0.3 -0.6 nM) binding site with a modest AII receptor density (Bmax range of 17-120 fmol/mg protein) comparable to rat lung (130 fmol/mg protein). White adipose tissue contained a greater number of AII receptor sites than brown adipose tissue. Competition displacement studies demonstrated the AT1 receptor is the only angiotensin receptor subtype localized in adipose tissue, with the rank order for competition of [125I]AII binding in all adipose tissues examined AIII > AII > losartan > angiotensin I (AI) > PD123319. The AT2 specific receptor antagonist, PD123319, was ineffective at displacing [125I]AII binding in all adipose tissues examined. Since components of the renin-angiotensin system are regulated in adipose tissue, we determined if the AII receptor is also regulated in the obese state. AII receptor binding characteristics were determined in liver, lung, ISBAT and EF membrane preparations from adult Zucker obese (fa/fa) and lean (Fa/?) rats. AII receptor density was decreased in liver from obese rats. In contrast, the affinity for [125I]AII binding was not altered in tissues from obese rats. In a separate group of obese and lean rats, regulation of the AII receptor by phenobarbital (PB) was examined. Administration of PB restored AII receptor density in liver from obese rats to levels obtained in lean rats. In summary, these results demonstrate the presence of AT1 receptor sites in brown and white adipose tissue. Moreover, AII receptor density is decreased in tissues from obese rats, with restoration of receptor density by administration of PB. Future studies will determine if PB regulates the AT1 receptor at the level of gene expression. PMID- 8726685 TI - Changes in angiotensin AT1 receptor density during hypertension in fructose-fed rats. AB - Feeding carbohydrate-enriched diets to normal rats has been shown to induce insulin resistance and hyperinsulinemia associated with an elevation of blood pressure. Previously we reported that the renin-angiotensin system (RAS) is likely to be involved in the elevation of blood pressure. The purpose of this study was to determine the changes in plasma angiotensin II (AII) and AII receptor density associated with the elevation of blood pressure in fructose treated rats. Male Sprague-Dawley rats were divided into two groups and were fed either normal rat chow or a 60% fructose-enriched diet for four weeks. Plasma insulin of fructose-treated rats was significantly elevated (p < 0.05) by the end of first week of fructose treatment and remained elevated throughout the study. Plasma AII levels of fructose-fed rats was 3.5 fold greater than the controls at the end of second week and returned to basal levels at the end of the fourth week of dietary treatment. Blood pressure was significantly elevated in the fructose fed rats within two weeks of fructose treatment. Elevation of blood pressure was associated with left ventricular hypertrophy. Angiotensin II type I receptor (AT1) density was determined in the left ventricle, aorta, adrenal gland and hypothalamus. There was a significant increase in AT1 receptor density in the ventricle at the end of third and fourth weeks of treatment, whereas there was a significant decrease in the receptor density in the aorta at the end of the fourth week of treatment. Receptor density in the adrenal gland and hypothalamus of fructose-fed rats was similar to their respective controls. The results of this study suggest that the RAS plays a role in the elevation of blood pressure of fructose-fed rats and also contributes to the ventricular hypertrophy observed in these rats. PMID- 8726686 TI - Cardiac effects of AII. AT1A receptor signaling, desensitization, and internalization. AB - Angiotensin II receptors present in cardiomyocytes, nonmyocytes (predominantly fibroblasts), nerve terminals, and the heart vasculature mediate the multiple actions of angiotensin II (AII) in the heart, including modulation of normal and pathophysiological cardiac growth. Although the cellular processes that couple AII receptors (principally the AT1 subtype) to effector responses are not completely understood, recent studies have identified an array of signal transduction pathways activated by AII in cardiac cells. These include: the stimulation of phospholipase C which results in the activation of protein kinase C and the release of calcium from intracellular stores; an enhancement of phosphaditic acid formation; the coupling to soluble tyrosine kinase phosphorylation events; the initiation of the mitogen activated protein kinase (MAPK) cascade; and the induction of the STAT (Signal Transducers and Activators of Transcription) signaling pathway. It is tempting to speculate that these latter responses, which have been previously associated with growth factor signaling pathways, are involved in AII-induced cardiac growth. Interestingly, some of these novel pathways are apparently not under the same strict control imposed upon the more classical signaling pathways. Thus, while AII-induced calcium transients are rapidly (within minutes) desensitized following exposure to AII, the MAP kinase pathway is not, and activation of the STAT pathway requires hours of agonist exposure for maximal induction. These observations support an emerging picture in which the downstream signal transduction pathways of AII receptors are initiated and terminated with a distinct temporal arrangement. This organization allows appropriate rapid responses (e.g. vascular contraction) to transient AII exposure, some of which are rapidly terminated, perhaps for protective reasons, and others not. In contrast, additional responses (e.g. growth) probably require prolonged exposure to agonist. PMID- 8726687 TI - AT1-receptors and cellular actions of angiotensin II in neuronal cultures of stroke prone-spontaneously hypertensive rat brain. AB - AT1-receptors, its mRNA and cellular actions of angiotensin II (Ang II) have been compared between neuronal cultures of Wistar Kyoto (WKY) and stroke-prone spontaneously hypertensive (SP-SH) rat brains. Bmax for AT1-receptor binding is 2 fold higher and is associated with a parallel increase in the levels of AT1 receptor mRNA in SP-SH rat brain neurons compared with WKY rat brain neurons. Ang II causes stimulation of both c-fos and norepinephrine transporter (NET) mRNAs in both strains of neurons and this stimulation is also 2-3-fold higher in SP-SH rat brain neurons compared with WKY rat brain neurons. In contrast, Ang II stimulation of PAI-1 mRNA in SP-SH neurons is only 50% that of in WKY rat brain neurons suggesting that SP-SH neurons express a decrease in AT1-receptor coupling with PAI-1 response. These observations demonstrate that SP-SH neurons express AT1-receptor-functions similar to those described for SHR neurons. PMID- 8726688 TI - Antisense oligonucleotides for in vivo studies of angiotensin receptors. PMID- 8726689 TI - Interactions of angiotensin II with central dopamine. AB - There is a large body of evidence to support the concept of a relationship between brain Ang II and catecholamine systems. This interaction may participate in some central actions of Ang II such as cardiovascular control, dipsogenesis, and complex behaviours. It also extends to the nigrostriatal dopaminergic system which bear AT1 receptors, both on their cell bodies in the substantia nigra presynaptically, and on their terminals in the striatum, where Ang II can markedly potentiate DA release. This observation suggests that drugs which modulate central Ang II may be useful in regulating central dopaminergic activity. PMID- 8726690 TI - Regulation of the hypothalmic-pituitary-adrenal axis and vasopressin secretion. Role of angiotensin II. PMID- 8726691 TI - Relationship between the drinking response to angiotensin II and induction of Fos in the brain. PMID- 8726692 TI - Identification of AT1 receptors on cultured astrocytes. PMID- 8726693 TI - Structure-activity relationship of the agonist-antagonist transition on the type 1 angiotensin II receptor; the search for inverse agonists. PMID- 8726694 TI - Molecular cloning and expression of angiotensin II type 2 receptor gene. PMID- 8726695 TI - Molecular cloning of the human AT2 receptor. PMID- 8726696 TI - Molecular and functional characterization of angiotensin II AT2 receptor in neuroblastoma N1E-115 cells. PMID- 8726697 TI - Characterization of the AT2 receptor on rat ovarian granulosa cells. PMID- 8726698 TI - AT2 receptor expression in ovaries: a review. PMID- 8726699 TI - Heterogeneity of rat angiotensin II AT2 receptor. PMID- 8726700 TI - Heterogeneity of angiotensin type 2 (AT2) receptors. AB - Evidence continues to accumulate that strengthens the proposal of heterogeneity within both the AT1 and the AT2 receptor subtypes. Pharmacologic, biochemical and immunological studies of AT2 receptors expressed in N1E-115 cells strengthen the hypothesis of AT2 receptor heterogeneity. However, it is important to reassess these studies, especially in terms of how these results correlate with other reports of AT2 receptor heterogeneity. For example, AT2 receptor immunoreactivity was absent in some neuronal regions which have previously been proposed to express the AT2 receptor subtype. In particular, AT2 receptor staining was not seen in the inferior olive, a region which is known to express a high density of AT2 receptors. Upon first examination, these results were somewhat troubling. However, when compared with earlier reports, these results should not have been unexpected. For instance, Tsutsumi and Saaverdra previously have shown that AT2 receptors in the locus coeruleus are sensitive to the actions of guanine nucleotides, while AT2 receptors in the inferior olive are insensitive (21). These antisera were raised against a population of AT2 receptors which are sensitive to GTP gamma S and therefore, the lack of AT2 receptor staining in the inferior olive, as well as the presence of AT2 receptor immunoreactivity in the locus coeruleus, confirms and extends these earlier reports. In addition the AT2 receptors expressed in the locus coeruleus have been shown to be functionally distinct from AT2 receptors in the inferior olive. In this regard, Ang II has been shown to depress glutamate-induced EPSPs in the locus coeruleus, an effect which is mediated through the AT2 receptor (19). Conversely, AT2 receptors have been shown to increase the firing rate of neurons in the inferior olive (20). Collectively, these results would predict that staining should be absent in the inferior olive using these AT2-directed antisera. Indeed, in view of these earlier physiological and pharmacological studies, the presence of AT2 receptor immunoreactivity in the inferior olive would have been surprising. The most convincing example of AT2 receptor heterogeneity is the characterization of AT2 receptors present in N1E-115 cells. Separation of solubilized N1E-115 membranes by heparin-Sepharose chromatography generates two populations of AT2 receptors which are pharmacologically and biochemically distinct. In particular, CGP42112A was approximately 2 orders of magnitude more selective for Peak III AT2 receptors than was PD123319. Binding activity of Peak I and Peak III AT2 receptor populations also differed in their responses to GTP gamma S and DTT treatment. Lastly, the AT2-directed antisera, raised against the Peak I population of AT2 receptors, were not able to immunodetect the Peak III population of AT2 receptors in immunoblot analysis, or immunoprecipiatate AT2 binding activity from Peak III material. Pharmacological, biochemical and immunological analysis of the AT2 receptor clone isolated from N1E-115 cells revealed that it has the identical characteristics or properties of the Peak III receptor. The AT2 receptor isolated from N1E-115 cells exhibited a similar pharmacology as the Peak III AT2 receptor, in that CGP42112A was more effective at displacing 125I-Ang II binding activity than was PD123319. The AT2 receptor clone was also shown to be insensitive to the actions of GTP gamma S, as well as demonstrated increased binding activity in the presence of DTT, identical to the Peak III AT2 receptor. Lastly, immunoblot analysis of membranes prepared from COS-1 cells transfected with the AT2 receptor cDNA from N1E-115 cells did not demonstrate any immune-specific bands with the AT2-directed antisera. Characterization of an AT2 receptor cDNA isolated from N1E 115 cells reveals that this clone is identical to the Peak III type of AT2 receptor. PMID- 8726701 TI - Angiotensin II stimulates protein phosphatase 2A activity in cultured neuronal cells via type 2 receptors in a pertussis toxin sensitive fashion. AB - Recent studies have suggested a role for an inhibitory G protein (Gi) and protein phosphatase 2A (PP2A) in the angiotensin II (Ang II) type 2 (AT2) receptor mediated stimulation of neuronal K+ currents. In the present study we have directly analyzed the effects of Ang II on PP2A activity in neurons cultured from newborn rat hypothalamus and brainstem. Ang II elicited time (30 min-24 h)- and concentration (10 nM -1 microM)-dependent increases in PP2A activity in these cells. This effect of Ang II involved AT2 receptors, since it was inhibited by the AT2 receptor selective ligand PD123319 (1 microM), but not by the Ang II type 1 receptor antagonist losartan (1 microM). Furthermore, the stimulatory effects of Ang II on PP2A activity were inhibited by pretreatment of cultures with pertussis toxin (PTX) (200 ng/ml; 24 h) indicating the involvement of an inhibitory G-protein; and by cycloheximide (CHX) (1 microgram/ml; 30 min) indicating a requirement for protein synthesis. These effects of Ang II appear to be via activation of PP2A, since Western Blot analyses revealed no effects of this peptide on the protein levels of the catalytic subunit of PP2A in cultured neurons. In summary, these data suggest that PP2A is a key component of the intracellular pathways coupled to neuronal AT2 receptors. PMID- 8726702 TI - Functional aspects of angiotensin type 2 receptor. PMID- 8726703 TI - Angiotensin receptor heterogeneity in the dorsal medulla oblongata as defined by angiotensin-(1-7). PMID- 8726704 TI - Atypical (non-AT1, non-AT2) angiotensin receptors. PMID- 8726705 TI - Brain angiotensin II and related receptors: new developments. PMID- 8726706 TI - Receptors for (3-8) angiotensin in brain cells. AngIV binding in brain cells. PMID- 8726707 TI - Esoteric contact dermatitis. Part IV: Devastating contact dermatitis in India produced by American parthenium weed (the scourge of India). PMID- 8726708 TI - A dermatologic diary. Portrait of a practice: a family deals with death. PMID- 8726709 TI - Becker's melanosis. PMID- 8726710 TI - Pyoderma gangrenosum associated with cystic acne and hidradenitis suppurativa controlled by adding minocycline and sulfasalazine to the treatment regimen. AB - Pyoderma gangrenosum often presents a difficult therapeutic challenge. The case is described of a 42-year-old black man with the association of cystic acne, hidradenitis suppurativa, and seronegative arthritis with pyoderma gangrenosum. The pyoderma gangrenosum ulcers remained refractory to treatment until therapies aimed in part at the associated diseases were begun. Minocycline was given for treatment of cystic acne and hidradenitis suppurativa as well as pyoderma gangrenosum. Sulfasalazine was prescribed for seronegative arthritis as well as pyoderma gangrenosum. The combination therapy permitted healing of the pyoderma gangrenosum ulcers. PMID- 8726711 TI - Necrobiotic xanthogranuloma with paraproteinemia. AB - Necrobiotic xanthogranuloma (NXG) is a rare skin disease mimicking necrobiosis lipoidica diabeticorum clinically and histologically. However, its association with paraproteinemia, and the evolution of hematologic malignancies in some cases, established NXG as a separate clinicopathologic entity. A typical case with indolent course is described. PMID- 8726712 TI - Coping with psoriasis: a guide for counseling patients. AB - Psoriasis is a common, chronic, intractable hyperproliferative skin disease that affects 1 to 3 percent of the American population. This disease accounts for 4.8 percent of office visits for dermatologic reasons, third after warts and acne. Two British dermatologists, however, note that dermatologists tend to pay "more attention to the rash and its transient, occasional removal than to the problems of the patient in the community in relation to that rash." PMID- 8726713 TI - Treatment of pyoderma gangrenosum with methotrexate. AB - Pyoderma gangrenosum is an ulcerative skin condition of unknown cause. It is often refractory to treatment, requiring high dosages of immunosuppressive medications. A patient with idiopathic pyoderma gangrenosum was successfully treated with methotrexate and weaned off steroids for the first time in four years. The patient had received multiple skin grafts and failed topical treatment, rifampin, dapsone, azathioprine, and intralesional steroids. She required prednisone 60 mg/day for lesion healing. Methotrexate was given orally, with rapid response, allowing discontinuation of prednisone seven months after beginning therapy. There is evidence of neutrophil dysregulation in pyoderma gangrenosum. Methotrexate has been shown to decrease neutrophil migration and chemotaxis, suggesting a role for its use in pyoderma gangrenosum. PMID- 8726714 TI - Cutaneous reaction to vasopressin. AB - Vasopressin (Pitressin, 8-arginine vasopressin) is a potent vasoconstrictor of splanchnic arterioles. When administered by continuous intravenous infusion, it reduces portal blood flow and pressure and is used in the management of bleeding esophageal varices. We describe a purpuric and necrotic cutaneous reaction to vasopressin that occurred at locations distant from intravenous catheter sites, and we review previous reports of similar reactions. PMID- 8726715 TI - Perforating elastosis in necrobiosis lipoidica diabeticorum. AB - A 58-year-old diabetic woman with necrobiosis lipoidica diabeticorum demonstrated lesions with raised, prominent borders. These areas were found on biopsy to have transepidermal elimination of elastic fibers. We present this case as a unique finding and review the characteristics and possible mechanisms of transepidermal elimination. PMID- 8726716 TI - Squamous cell carcinoma of the nail bed. AB - Squamous cell carcinoma of the nail bed is a rare disorder. Metastasis is extremely unusual. A case of primary squamous cell carcinoma of the nail bed with metastasis to the regional lymph nodes is reported. The tumor was successfully treated by amputation of the involved finger, block dissection of the regional lymph nodes, postoperative irradiation of the involved axilla, and systemic chemotherapy. Early diagnosis by biopsy, especially in patients with recurrent and persistent disease of the nail bed, is recommended, so that treatment can be instituted early while the tumor is confined to its primary site. PMID- 8726717 TI - Rubinstein-Taybi syndrome with multiple flamboyant keloids. AB - A single case of Rubinstein-Taybi syndrome associated with numerous giant keloids is reported. The patient manifested all the major features of Rubinstein-Taybi syndrome. The cause of this multisystem developmental disorder is unknown. PMID- 8726718 TI - Traumatic eosinophilic ulcer of the oral mucosa: a misdiagnosed entity. AB - We report a case of and review the literature on eosinophilic ulcer of the tongue. This is a benign entity frequently unrecognized in clinical practice, with only occasional mention in the literature, the clinical signs of which are easily confused with those of squamous cell carcinoma. Although the histopathologic appearance of this entity is distinctive, its clinical course and rapid healing without therapy themselves constitute strong diagnostic clues, making diagnostic or therapeutic bloody methods generally unnecessary. PMID- 8726719 TI - A man with nevoid hyperkeratosis of the areola. AB - Nevoid hyperkeratosis of the nipple and areola is a unique clinical entity and a diagnosis of exclusion. The patient presenting with nipple/areolar hyperkeratosis must be examined carefully for other underlying cutaneous diseases such as epidermal nevi, ichthyosis, acanthosis nigricans, Darier's disease, cutaneous T cell lymphoma, and other chronic skin dermatitides that may be responsible for the changes. If no other clinical findings are evident, the diagnosis can be made. Although the disorder is more common in women of childbearing age, men may show nevoid changes after estrogen therapy or idiopathically. The use of topical 12 percent lactic acid lotion (Lac-Hydrin) resolves the skin changes over a six month period. PMID- 8726720 TI - Trichorrhexis nodosa: a manifestation of hypothyroidism. AB - A 23-year-old woman presented with diffuse white hair nodules, hair fragility, and the inability to grow long hair. Examination of specimens under the light microscope showed the morphologic characteristics of trichorrhexis nodosa. Results of tests revealed hypothyroidism. Treatment with a daily dose of 0.1 mg L thyroxine sodium for six months restored euthyroidism and concurrently normalization of the hair defect was observed. The corrected hair defect observed after replacement therapy may point toward a causal relationship between hypothyroidism and trichorrhexis nodosa. To the best of our knowledge, this is the first report of trichorrhexis nodosa associated with hypothyroidism. PMID- 8726721 TI - A retrospective evaluation of patch testing in patients diagnosed with allergic contact dermatitis. AB - One reason given by clinicians for not using patch testing in patients with dermatitis is that it may not improve the treatment outcome. The main objectives of this retrospective study were to determine the frequency of use of patch testing as an aid to the diagnosis of allergic contact dermatitis (ACD), to determine the ACD-related costs before and after diagnosis of the disease, and to determine the benefit of the diagnosis of ACD, with or without patch testing, to the patient's treatment outcome. This study will also provide definite direction for a prospective outcome study. Medical records of 270 patients with ACD were utilized retrospectively to study the benefit of patch testing. Patch testing was performed on 56 percent of these patients. Patch testing was found to bring about a larger decrease in the disease severity index and percentage disease activity from prediagnosis to postdiagnosis than diagnosis without patch testing. In the group of patients with a prediagnosis disease duration of two months to one year, the postdiagnosis duration was considerably less for those patch tested and treated than those that were treated without patch testing. Patch testing was found to be the most cost-effective in patients with a disease duration of two months to one year. PMID- 8726722 TI - Influence of slaughter weight on growth and carcass characteristics, commercial cutting and curing yields, and meat quality of barrows and gilts from two genotypes. AB - Pigs representing two crossbred genotypes, a breeding company hybrid (BCH) and a three breed cross (Hampshire x [Yorkshire x Duroc]) (HYD) were evaluated at five slaughter weights (SLW): 100, 115, 130, 145, and 160 kg. A total of 160 pigs were grown in like-sex (barrows [B] or gilts [G]), like-genotype groups, with four pigs/group, from 60 kg live weight. A corn-soybean meal-based diet was available on an ad libitum basis (15.8% crude protein, 3,300 kcal/kg ME). One-half of the pigs from each group (80 pigs) were slaughtered for carcass and meat quality evaluation. Genotype BCH grew faster, had lower backfat depths in the loin and lumbar regions, and a smaller loin eye area than HYD, but both groups had a similar gain:feed ratio. Few consistent genotype differences in cutting and curing yields and meat quality were observed. Differences between sexes for growth and carcass traits were generally in agreement with previous research; however, the magnitude of the differences was small. There were few nonlinear regressions involving SLW and limited differences between genotypes or sexes in the slopes of the linear regressions. Increases in SLW were associated with increases in feed intake, backfat depth and loin' eye area, and minimal changes in growth rate or gain:feed. Percentage of loin increased and ham, shoulder, and spare rib percentages decreased with slaughter weight. The weight of trimmed, boneless cuts increased with slaughter weight, but percentage trimmed, boneless cuts was reduced. Curing yields for belly increased with slaughter weight. Changes in meat quality with increasing slaughter weight were relatively small. Longissimus lumborum fat content increased and moisture content decreased with slaughter weight. These results suggest that modern genotypes can be slaughtered at live weights up to 160 kg with limited impact on growth performance, commercial meat yields, or meat quality characteristics. PMID- 8726723 TI - The growth performance, carcass characteristics, and meat quality of halothane carrier and negative pigs. AB - Growth performance, carcass characteristics, and meat quality of halothane carrier (Nn) and negative (NN) pigs grown from 40 kg live weight and slaughtered at three weights (110, 125, and 140 kg live weight) were studied. Daily gains were similar for the two genotypes (974 g for Nn and 964 g for NN), but Nn pigs had a higher gain:feed ratio than NN pigs (P < .01). Dressing percentage was higher in Nn pigs than in NN pigs (P < .001), but there were no genotype differences for carcass length, backfat thickness, or loin eye area. Percentage yield of trimmed, boneless wholesale cuts was higher for Nn pigs than for NN pigs (P < .05). This resulted from higher trimmed, boneless ham, boston, and picnic weights (P < .05) in Nn than in NN pigs (6.9, 3.2, 3.5 kg vs 6.6, 3.0, 3.4 kg, respectively). The weight of fat-free lean was higher in Nn pigs (P < .05). The longissimus thoracis muscle from carrier pigs had lower 45 min (P < .001) and 24 h (P < .01) pH. Longissimus lumborum samples from carriers had lower (P < .001) subjective' meat quality scores and a higher drip loss (P < .001); however, cooking loss, eating quality, and shear force values were similar for the two genotypes. There were no important slaughter weight x genotype interactions for the traits reported. Overall, the data from this study suggest Nn pigs had an advantage over NN pigs in terms of feed efficiency, carcass yield fat-free lean content, and commercial lean cut yields but had a higher incidence of PSE. PMID- 8726724 TI - Relationship of weaning-to-estrus interval to timing of ovulation and fertilization in sows. AB - Data from two experiments were used to study the relationship between weaning-to estrus interval (WEI, varying from 3 to 6 d) and the duration of estrus, the interval from onset of estrus to ovulation and fertilization results at 120 h after ovulation, in 201 multiparous sows. An increase in WEI resulted in a decrease in the duration of estrus (Duration of estrus (h) = [81 (exp1) or 88 (exp2)]-.33 x WEI, R2 = .25, P < .001) and a decrease in the interval from onset of estrus to ovulation (onset estrus to ovulation (h) = [54 (exp1) or 59 (exp2)] .20 x WEI, R2 = .23, P < .001). Sows that were inseminated between 0 and 24 h before ovulation showed consistent high fertilization results (91% normal embryos), irrespective of WEI. Sows that were inseminated outside this range showed an increased proportion of unfertilized eggs (P < .001). This increase occurred similarly in all weaning to estrus intervals. These results suggest that negative effects of a longer WEI on subsequent litter sizes and farrowing rates as described in literature might be a result of suboptimal timing of insemination relative to ovulation and not of poor fertility in these sows. PMID- 8726725 TI - Testes function and feedlot performance of bulls actively immunized against gonadotropin-releasing hormone: effect of age at immunization. AB - Testes function, feedlot performance, and carcass traits were evaluated in bulls actively immunized against gonadotropin-releasing hormone (GnRH) at different ages. Bull calves were randomly assigned to one of seven treatment groups (n = 15 calves/group). Calves were unimmunized (Group 1), immunized at 1.5, 4, 7, or 12 mo of age with a GnRH-keyhole limpet hemocyanin (KLH) conjugate (Groups 2 to 5, respectively), or castrated at 4 mo of age (Groups 6 and 7). Immunized bulls did not receive a secondary, or booster, immunization. Calves in group 6 received Synovex-C at castration and Synovex-S at weaning and feedlot entry. Anti-GnRH titer was evident at slaughter in all immunized bulls. However, the final immune response of bulls immunized at 1.5 mo was significantly lower than the response of bulls immunized at later stages of development. Final scrotal circumference and testis weight in bulls immunized at 4, 7, or 12 mo of age were significantly reduced relative to unimmunized bulls. The final live weight, feedlot gain, and carcass weight of immunized and unimmunized bulls did not differ (P > .05) from the same parameters in steers implanted with Synovex. Longissimus muscle area, marbling score, and backfat thickness did not differ between immunized and unimmunized bulls. The sex class score of the carcasses of immunized bulls did not differ from the score of steer carcasses. In contrast, a significantly higher proportion of carcasses from unimmunized bulls graded as bullock carcasses. Taken together, these data indicate that a single immunization against GnRH at 4 to 12 mo of age results in significant attenuation of testicular growth in bulls. These data also demonstrate that immunization against GnRH reduces the masculinity of carcasses from bulls, but does not affect feedlot performance, longissimus muscle area, marbling score, or backfat thickness. These results suggest that single immunization with the GnRH-KLH conjugate may have practical utility as a noninvasive alternative to surgical castration in management of beef cattle. PMID- 8726726 TI - Evaluation of F1 calves sired by Brahman, Boran, and Tuli bulls for birth, growth, size, and carcass characteristics. AB - Birth (n = 308), weaning (n = 291), feedlot and carcass (n = 142), and yearling heifer traits (n = 139) were evaluated in F1 calves sired by Brahman (BR), Boran (BO), and Tuli (TU) bulls and born to multiparous Hereford and Angus cows. Calves sired by BR were heaviest (P < .05) at birth and largest (P < .05) for cannon bone length. Tuli crosses were smaller (P < .05) for birth weight and cannon bone length than BR and BO crosses. No significant differences were observed for gestation length among sire breeds. Brahman crosses had larger (P < .05) weaning weight, hip height, and preweaning ADG than calves sired by BO and TU. Similar trends were observed for feedlot traits. Carcasses of BR crosses were heavier (P < .05) and had less (P < .05) internal fat than those of BO and TU crosses. Tuli crosses averaged greater (P < .05) skeletal maturity than BO crosses. Tuli crosses averaged greater marbling (P < .05) than BR crosses and less (P < .05) Warner-Bratzler shear force than BO crosses. No differences were observed in longissimus muscle area, fat thickness, or yield grade among sire breeds. Heifers sired by BR were heaviest (P < .05) and tallest (P < .05) at yearling measurement. Brahman F1 heifers had larger (P < .05) pelvic height and pelvic area, due to larger skeletal frame size, than BO and TU F1 heifers. These results indicate large differences in growth and skeletal size exist among calves sired by these three breeds. Several important differences also exist for carcass quality traits, but not for carcass yield traits, among these three breeds. PMID- 8726727 TI - Persistence of the effects of early experience on consumption of low-quality roughage by sheep. AB - The aim of this study was to determine the effects of experience early in life with cured weeping lovegrass (Eragrostis curvula) (low-quality roughage) or fresh oats (Avena sativa) (high-quality roughage) on voluntary intake and nitrogen retention when sheep ate a low-quality roughage 9 mo after initial exposure. From 1 to 5 mo of age, experienced wethers (EW) grazed cured weeping lovegrass, whereas inexperienced wethers (IW) grazed fresh oats (initial exposure). Then both EW and IW were fed sorghum (Sorghum bicolor) hay for 45 d, fresh weeping lovegrass for 105 d, and finally alfalfa hay for 120 d. Nine months after completion of the initial exposure EW and IW were offered sorghum hay (low quality roughage), and, in addition, both groups were offered sorghum with ad libitum access simultaneously to alfalfa hay at six levels of availability. Experienced wethers ingested 15% more (P < .02) sorghum than IW. When given sorghum simultaneously with alfalfa at different levels of availability, EW ingested more (P < .10) sorghum than IW when only sorghum was freely available. Nitrogen retention was negative in both groups, but it was less negative in EW than in IW per unit of metabolic weight (P < .09) and per wether (P < .04). The results show that early dietary experience can have profound and persistent effects on consumption of foods low in nutritional quality, apparently through changes in critical physiological mechanisms. PMID- 8726728 TI - Dairy calves' discrimination of people based on previous handling. AB - To determine whether calves can distinguish between different people, we examined their contact with familiar and unfamiliar people and with people who handled them positively or aversively. When a familiar or unfamiliar person entered the calves' pens, latency to contact and duration of contact were the same, but bouts of contact were shorter and more frequent if the person was unfamiliar. In Exp. 2, calves were treated repeatedly in their home pens, by three handlers: one treated them positively, one aversively, and a third (neutral) did not interact with them. After seven treatments, calves tended to avoid all three handlers. After 12 treatments, calves contacted the positive handler significantly more than the aversive handler in their home pens. However, when retested outside their home pens, the calves did not discriminate. In Exp. 3, calves were treated positively and aversively in a novel treatment pen rather than in their home pens. The calves made more contact with the positive handler than the aversive handler after seven treatments. When retested in their home pens, most of the calves continued to avoid the aversive handler, but some made contact with the aversive handler. Calves can readily discriminate between different people based on their previous experience. They can develop a general fear of people as a result of aversive handling, and positive handling is required to overcome this. However, some calves do not generalize their fear of an aversive handler to places other than those in which they were handled. PMID- 8726729 TI - Depression of lipogenesis in swine adipose tissue by specific dietary fatty acids. AB - The objective of this study was to document the influence of specific dietary fatty acids on rates of lipid synthesis and sensitivity to insulin in porcine adipose tissue. Weanling pigs were assigned to one of six groups, and each group was fed diets containing 10 g/100 g of added cornstarch or 10 g/100 g of added fatty acid. The fatty acid-enriched diets contained either a combination of 14:1 plus 16:1 (14:1/16:1 diet), 16:0, 18:0, 18:1, or 18:2 (n-6). With the exception of the cornstarch diet, all diets contained approximately 35% 14:0. Subcutaneous adipose tissue samples were collected at slaughter from the area overlying the first cranial vertebra. Fresh samples were incubated for 2 h in 20 mM glucose and 0, 10, 100 or 1,000 microU/mL of porcine insulin. The smallest adipocytes were observed in adipose tissue from pigs fed the 16:0 or 18:2 diets. Glucose incorporation into lipids was greater (P < .05) in adipose tissue from cornstarch fed pigs than in adipose tissue from the other treatment groups. Lipogenesis was 67, 53, 35, 32, and 20% lower (P < .05) in adipose tissue from 16:0-, 14:1/16:1-, 18:0-, 18:2-, and 18:1-fed pigs, respectively, than in adipose tissue from the cornstarch-fed pigs. Insulin increased lipogenesis by 19% (P < .05) in adipose tissue from the cornstarch-fed pigs and by 15 to 40% (P < .05) in adipose tissue from the 14:1/16:1-fed pigs. Insulin did not stimulate lipogenesis (P > .4) in adipose tissue from pigs fed the 16:0, 18:0, or 18:1 diets. The data suggest that fatty acid chain length and unsaturation are determinants in the effects of dietary fat and insulin on de novo lipogenesis. PMID- 8726730 TI - Evidence of classic beta 3-adrenergic receptors in porcine adipocytes. AB - Adipocytes from several mammalian species have predominant beta 3-adrenergic receptors (beta 3-AR). Attempts to classify porcine adipocyte beta-adrenergic receptors (beta-AR) into subtypes have not been successful. Selectivity of agonists and antagonists for stimulation of lipolysis and for ligand binding is considerably more restrictive than for the classic rat and guinea pig beta-AR subtypes. The unique pattern for activity of agonists and antagonists in porcine beta-AR precludes analogy to classic receptors and consequently there is no conclusive evidence regarding porcine beta-AR subtypes. Porcine adipocyte membranes were used in ligand binding experiments designed specifically to demonstrate beta 3-AR. Equilibrium saturation curves with dihydroalprenolol, CGP 12,177, or iodocyanopindolol indicated saturation at low concentrations with a single binding site. Equilibrium competitive ligand binding with iodocyanopindolol as radioligand and isoproterenol or propranolol as competitive ligands indicated both ligands totally inhibited radioligand binding; propranolol was more potent than isoproterenol. Nonradioactive CGP 12,177 also competed with iodocyanopindolol. Ligand binding experiments provided no evidence of a low affinity beta-AR (binding at high concentrations of ligand), the beta 3-AR. Positive evidences of a beta 3-AR were that CGP 12,177, a beta 1- and beta 2 adrenergic receptor antagonist but a beta 3-AR agonist, partially stimulated porcine adipocyte lipolysis. Furthermore, transcripts for a beta 3-AR, as well as a beta 1- and beta 2-AR, have been demonstrated in RNA from porcine adipocytes in other studies. The beta-AR subtypes expressed and functional in porcine adipocytes remain unknown. The multiple ligand binding sites cannot be attributed to classic beta-AR subtypes. The porcine beta-AR may be a single unique receptor to impart atypical binding properties, or more likely, multiple subtypes, each different enough from classic subtypes to impart the unique properties observed. PMID- 8726731 TI - Proteolysis of specific muscle structural proteins by mu-calpain at low pH and temperature is similar to degradation in postmortem bovine muscle. AB - Postmortem (PM) and mu-calpain-induced degradation of specific skeletal muscle proteins was monitored by SDS-PAGE and Western blotting. Samples were removed from bovine longissimus thoracis (LT) at approximately 45 min PM for the preparation of at-death (0-d) myofibrils (MF). The LT was excised at 1 d PM, vacuum-packaged, and stored at 2 degrees C. Samples were removed for Warner Bratzler shear force analysis and biochemical analysis at 1, 3, 7, 14, 28, and 56 d PM. The protease mu-calpain was purified from bovine skeletal muscle and used to digest at-death MF at pH 5.6, 4 degrees C, 100 microM CaCl2. Degradation of the proteins titin, nebulin, filamin, desmin, and troponin-T was monitored in the PM and mu-calpain-digested samples by using SDS-PAGE and Western blotting. The PM samples that had significantly lower shear force (LSF) values (P < .05) at 1 d PM exhibited faster degradation of these five proteins than the higher shear force (HSF) samples. In LSF samples, the intact titin band (T1) was absent by 7 d PM and nebulin was absent by 3 d PM. In LSF samples, some filamin was degraded by 3 d PM, but in HSF samples degradation was not apparent until 14 d PM. In LSF samples, desmin was degraded more rapidly PM than in HSF samples. Troponin-T was broken down PM to yield two major polypeptides of approximately 28 and 30 kDa; these polypeptides appeared earlier PM in LSF samples. Degradation products, similar to those observed PM, for all five proteins also were detected in Western blots of mu-calpain-digested MF, suggesting the calpain system plays a key role in PM protein degradation. PMID- 8726732 TI - Use of expected progeny differences for marbling in beef: I. Production traits. AB - Six Angus bulls with HIGH ( > .4) and six bulls with LOW ( < -.16) expected progeny differences (EPD) for marbling were used to evaluate the impact of marbling on progeny production and carcass traits. Bulls were randomly bred to MARC II (1/4 Hereford, 1/4 Simmental, 1/4 Angus, 1/4 Gelbvieh) composite cows in each of 2 yr to calve in the spring. At weaning, steers and heifers were separated and managed in different production systems. Steers (n = 131) were fed a growing diet (1.1 Mcal of NEg/kg) for 48 d followed by adaptation to a 93% concentrate finishing diet. Heifers (n = 125) were fed a growing diet (.79 Mcal of NEg/kg) for 191 d followed by adaptation to the same 93% concentrate diet. Steers and heifers from each treatment were slaughtered at two times spaced about 60 d apart within both years. Marbling EPD class had no effect on fat thickness, USDA yield grade, carcass weight, finishing daily gain, finishing DMI, or finishing efficiency (P > .18). More (P < .05) carcasses of calves from sires with HIGH EPD for marbling graded USDA Choice than from LOW EPD sires, 74% vs 47%, respectively. Angus sires can be selected to produce progeny that have increased ability to grade Choice without increasing yield grade or decreasing animal growth or feed efficiency. PMID- 8726734 TI - Characterization of biological types of cattle (Cycle IV): carcass traits and longissimus palatability. AB - Carcass and longissimus thoracis palatability traits from 888 steers obtained from mating Hereford and Angus cows to Hereford or Angus (HA), Charolais (Ch), Gelbvieh (Gb), Pinzgauer (Pz), Shorthorn (Sh), Galloway (Gw), Longhorn (Lh), Nellore (Ne), Piedmontese (Pm), and Salers (Sa) sires were compared. Data were adjusted to constant age (426 d), carcass weight (324 kg), fat thickness (1.2 cm), fat trim percentage (23%), and marbling (small00) end points. At a constant age of 426 d, carcasses from Ch steers were heaviest (P < .05) and Gw and Lh carcasses were lightest (P < .05). Adjusted fat thickness was greatest (P < .05) on carcasses from HA and least (P < .05) on carcasses from Ch, Gb, Lh, and Pm steers. USDA numerical yield grades were lowest (P < .05) for carcasses from Pm and highest (P < .05) for carcasses from HA, Ne, and Sh steers. Marbling scores were highest (P < .05) for carcasses from HA, Pz, and Sh and lowest (P < .05) for carcasses from Ch, Ne, and Pm steers. Longissimus thoracis from Pz had a lower (P < .05) shear force than that from all other breeds except HA, Gb, and Pm. Longissimus thoracis of carcasses from Ne steers was least (P < .05) tender. Adjustment of traits to various end points resulted in some changes in sire breed differences depending on the end point and the trait being considered but had little effect on palatability traits. Carcasses from Pm-sired steers provided the most desirable combination of carcass and longissimus palatability traits. PMID- 8726733 TI - Use of expected progeny differences for marbling in beef: II. Carcass and palatability traits. AB - A 2-yr study was conducted to determine the effect of EPD for marbling on marbling score, palatability traits, and carcass fatness in beef. Steer (n = 122) and heifer (n = 123) carcasses were obtained by mating Angus bulls having a high ( > .4) or low ( < -.16) EPD for marbling to MARC II cows (1/4 Angus, 1/4 Hereford, 1/4 Simmental, and 1/4 Gelbvieh). Carcass traits, composition of primals, quarters, and sides, palatability, and shear force data were obtained and adjusted to the mean number of days on feed, equal marbling score (Small50), fat thickness (1.0 cm), and carcass weight (318 kg) end points. Steer carcasses from the high marbling EPD group, adjusted to the mean number of days on feed, had significantly more marbling (P < .01) and less subcutaneous fat in the side and the hindquarter (P < .10) than their low marbling EPD counterparts. Adjusting steers to Small50 marbling produced smaller longissimus muscle area (by 5 cm2), less fat thickness (1.15 vs 1.28 cm), and lighter side weights (306 vs 333 kg) for high marbling vs low marbling EPD groups, indicating a faster rate of marbling deposition. Similar relationships of a greater magnitude were found for heifers, perhaps because the heifers were older than the steers at slaughter. No differences in taste panel ratings or shear force values were noted among steer carcasses. Heifer carcasses from the high marbling EPD group had better (P < .05) ratings for juiciness, muscle fiber tenderness, and overall tenderness than the low marbling EPD group heifers. These results indicate that it is possible, using existing genetic resources, to maintain marbling score and decrease fat in other depots of the carcass without compromising palatability. PMID- 8726735 TI - Influence of imposed feed intake patterns during lactation on reproductive performance and on circulating levels of glucose, insulin, and luteinizing hormone in primiparous sows. AB - Thirty-six primiparous sows were assigned to one of five treatments designed to mimic the lactation feed intake patterns observed in a previous study of commercial farms: high energy intake (HHH) or low energy intake during each week of a 3-wk lactation (LLL), or reduced intake during wk 1 (LHH), wk 2 (HLH) or wk 3 of lactation (HHL). The metabolizable energy intake of sows was either 16.5 (H) or 6.5 (L) Mcal/d. Diets were equal in lysine, providing 45 g/d. The HHH sows had a shorter (P < .05) weaning-to-estrus interval (9 +/- 3.2 d) than the sows in the LLL (23 +/- 3.5 d), HLH (22 +/- 3.5 d), and HHL (18 +/- 3.2 d) groups. The LH pulse frequencies of HHH (d 14: 1.5 pulses/8 h; d 21: 2.1 pulses/8 h) were greater (P < .05) than those of LLL (.2 pulses/8 h) and HLH (.5 pulses/8 h) on d 14 and those of LLL (0 pulses/8 h) and HHL (.9 pulses/8 h) on d 21 of lactation. No differences (P > .1) in mean serum concentrations of LH were observed among treatment groups either on d 21 of lactation or on d 1 postweaning. When fed a low-energy diet, sows had lower (P < .05) concentrations of serum insulin and plasma glucose than did sows fed a high-energy diet. Our results indicate that energy intake during lactation influences circulating insulin and glucose levels and LH pulse frequency and amplitude during midlactation and during the postweaning period. PMID- 8726736 TI - The influence of dietary salt level on water consumption, farrowing, and reproductive performance of lactating sows. AB - During two consecutive lactations using a switch-back design, 16 sows received a corn, cassava, barley, and soybean meal diet supplemented with either 8.5 or 1 g/kg of NaCl (.4% Na or .1% Na, respectively). The average daily water consumption during the 4-wk lactation period was greater (P < .01) for the sows fed the high-salt diet than for those fed the low-salt diet (13.9 vs 12.4 L, respectively). Urine production accounted for 27% of the water intake during lactation. This resulted in a difference during the entire lactation period of 42.2 L in water consumption (P = .07) and 11.4 L in urine production (P = .07). The Na and Cl concentrations of the urine for sows fed the high-salt diet were higher than for sows fed the low-salt diet (P < .05), but the K, Mg, P, and Ca concentrations were not influenced. The milk composition, measured at d 1, 13, and 27 of lactation, was relatively independent of the dietary salt level, except for the Cl concentration at d 1 after farrowing, which was higher (P < .05) for sows fed the high-salt diet. The moisture and ash content of the milk increased with the length of the lactation, irrespective of the diet. The increased ash content on d 13 and 27 of lactation was accounted for by the increased concentrations of P and Ca. There was no effect of dietary salt level on weight change of the sow or on the growth performance of the pigs. The interval from weaning to estrus (n = 13, t = 2.48, P = .029) was shorter for sows fed the high salt diet than for sows fed the low-salt diet (6.2 +/- 1.0 and 12.6 +/- 3.1 d, respectively). More sows were unsuccessfully mated after receiving the low-salt diet during lactation. These results indicate that a low-salt diet for lactating sows results in lower water consumption, lower urine production, and lower Na and Cl concentrations in the urine. Milk composition, weight change of the sow, and growth performance of the pigs were not altered, but reproductive failures afterward may point to a salt deficiency during lactation. PMID- 8726737 TI - Lysine requirement of the lactating sow determined by using plasma urea nitrogen as a rapid response criterion. AB - The dietary lysine requirement of adult sows nursing 10 pigs was estimated by using plasma urea nitrogen (PUN) concentration as a rapid response criterion. A preliminary experiment indicated that a 3-d feeding time is required to re equilibrate PUN concentrations after a change in the dietary concentration of lysine. In the main experiment, 12 sows (BW = 219 +/- 5 kg; parity = 4.5 +/- .3; ultrasound backfat = 21.3 +/- .9 mm) were used in two 6 x 6 Latin squares. Treatments consisted of dietary lysine intakes (30.2, 36.9, 43.6, 50.3, 57.0, and 63.7 g/d) that were assigned to six 4-d periods and to six sows in each square. Diets provided adequate levels of all nutrients other than lysine. The treatment periods started on d 5 of lactation and ended on d 29. Plasma urea N concentrations were determined on the 3rd and 4th d of each of the six 4-d treatment periods. Plasma urea N concentrations decreased quadratically (P < .02) with, increasing dietary lysine intakes. A two-slope, broken-line regression model was used to estimate the dietary lysine intake that minimizes PUN concentrations. The results indicated that adult sows nursing 10-pig litters with an average growth of 2.22 kg/d required 55.3 g/d of dietary total lysine to minimize PUN concentrations and, therefore, presumably to minimize body protein mobilization. PMID- 8726738 TI - Efficacy of a lysine-tryptophan blend for growth of chicks. AB - Two chick experiments were conducted to compare the growth-promoting efficacy as well as the toxicity of a new source of L-tryptophan and L-lysine, Tryptosine (16.1% tryptophan, 56.3% lysine). A corn-feather meal-soybean meal basal diet was made singly deficient in either lysine or tryptophan, and graded doses of lysine or tryptophan from either Tryptosine or feed-grade sources of lysine and tryptophan were supplemented. Linear (P < .01) weight gain responses occurred, and responses to lysine or tryptophan in Tryptosine were similar to those obtained with equal doses of lysine or tryptophan provided by feed-grade sources of L-lysine.HCI or L-tryptophan. The toxicity trial involved additions of 1, 2, or 4% lysine with .29, .58, or 1.16% tryptophan to a lysine- and tryptophan adequate corn-soybean meal diet. Both amino acids were provided as either Tryptosine or as feed-grade sources of lysine and tryptophan. Weight gain and feed intake were reduced in a linear fashion (P < .01) as levels of the two excess amino acids increased. The decreases caused by Tryptosine were similar to those caused by equivalent levels of excess feed-grade lysine and tryptophan. PMID- 8726739 TI - The role of excitatory amino acids in pulsatile secretion of luteinizing hormone in gilts and barrows. AB - The relationship of excitatory amino acid (EAA) activity to LH secretion was investigated in ovariectomized crossbred prepuberal gilts (93 +/- 1 kg BW) and Yorkshire barrows (94 +/- 2 kg BW) in two experiments. In Exp. 1, eight gilts received, i.m., saline (S) or 20 mg of Ketamine (K)/kg BW, a noncompetitive EAA receptor antagonist. Within these groups, four then received 10 mg of N-methyl-DL aspartate (NMA)/kg BW, an EAA agonist, or S i.v. Mean serum LH concentrations were similar among groups before treatment, did not change after S+S, but decreased (P < .05) by 1 h after S+NMA, 3 h after K+S, and 2 h after K+NMA. Serum cortisol concentrations did not change after S+S, but were increased (P < .05) from 30 to 90 min after S+NMA, at 120 min after K+S, and from 30 to 120 min after K+NMA. In Exp. 2, barrows received 2.5 mg of NMA/kg BW i.v. immediately after i.m. injection of S (n = 7) or 19.9 mg of K/kg BW (n = 8). Mean serum LH concentrations for the 2 h before treatment were similar among barrows, but decreased (P < .05) by 2 h after K+NMA and was not altered after S+NMA. Serum cortisol concentrations were increased at 30 min after S+NMA and from 60 to 90 min after K+NMA. We suggest that EAA both inhibit and stimulate LH secretion, with the inhibitory effects lying within the basal hypothalamus and the stimulatory effects lying within higher brain centers. PMID- 8726740 TI - Differential response of the luteal phase and fertility in cattle following ovulation of the first-wave follicle with human chorionic gonadotropin or an agonist of gonadotropin-releasing hormone. AB - A series of experiments with Holstein heifers was conducted to develop the capability of inducing accessory corpus luteum (CL) with a GnRH agonist (Buserelin, 8 micrograms; GnRHa) or hCG (3,000 IU) to increase plasma progesterone concentrations (Exp. 1, 2, and 3) and to test whether induction of accessory CL with hCG will increase conception rates in heifers (Exp. 4) and lactating cows (Exp. 5). In Exp. 1, heifers were treated on d 5 after estrus with GnRHa (n = 8) or saline (n = 7); heifers in Exp. 2 received hCG (n = 5) or saline (n = 4) on d 5. Experiment 3 allowed a contemporary evaluation of heifers treated on d 5 with GnRHa (n = 6), hCG (n = 6), saline (n = 6), or GnRHa at d 5 and hCG at the time of the induced ovulation (n = 5). The GnRHa and hCG were equally effective in inducing an accessory CL (93% induction rate), but the subsequent increase in progesterone concentrations was greater in hCG-treated heifers. A greater half life of hCG may provide longer LH-like stimulation of the first-wave follicle and subsequent developing accessory CL or a greater luteotropic effect on the original CL. Induction of an accessory CL with hCG on d 5 or 6 after insemination did not increase pregnancy rates in fertile heifers (Exp. 4: hCG = 64.8% vs control = 62.9%; n = 243) or lactating dairy cows during summer heat stress (Exp. 5: hCG = 24.2% vs control = 23.5%; n = 201). PMID- 8726741 TI - Use of a gonadotropin-releasing hormone agonist or human chorionic gonadotropin for timed insemination in cattle. AB - Three experiments were conducted to evaluate a synchronization protocol with AI at a predetermined time. In Exp. 1, 169 dairy heifers were assigned randomly to two groups: 1) timed AI (TAI), consisting of GnRH agonist injection (d 0, 1700), PGF2 alpha injection (d 7, 1700), GnRH agonist injection (d 8, 1700), and AI (d 9, 0800); and 2) AI at estrus (AIE), consisting of GnRH agonist injection (d 0, 1700), PGF2 alpha injection (d 7, 1700), and AI at detected estrus. Pregnancy rate was 25.8% for TAI (n = 89) compared with 48.7% for AIE (n = 80; P < .001). Experiment 2 was comparable to Exp. 1, but the second GnRH agonist injection in TAI was given 48 h after injection of PGF2 alpha. Heifers in TAI (n = 187) were inseminated at detected estrus if estrus occurred within 39 h after administration of PGF2 alpha (n = 47). Pregnancy rates were 45.5% for TAI and 48.0% for AIE (n = 177). Conception rate was reduced for TAI (45.5 [85/187] < 61.2% [85/139]; P < .005). In Exp. 3, the second injection of GnRH agonist, given at 48 h after injection of PGF2 alpha, was replaced with hCG (3,000 IU, i.m.). No differences in pregnancy rate were detected for TAI (52.9% [54/102]) vs AIE (56.1% [55/98]). Conception rate was reduced for TAI (52.9 [54/102] < 72.3% [55/76]; P < .005). Delaying the second GnRH agonist injection by 24 h improved pregnancy rate, but replacing the second injection of GnRH agonist with an injection of hCG did not prevent a reduction in conception rate. PMID- 8726742 TI - Ovarian response after gonadotropin treatment of heifers immunized against gonadotropin-releasing hormone. AB - Angus x Hereford heifers were used to determine whether the ovaries of heifers immunized against GnRH would respond to exogenous gonadotropins. Heifers (n = 23; 24 mo; 491 +/- 23 kg) were assigned to a 2 x 2 factorial experiment: immunization against GnRH conjugated to human serum albumin (GnRH-IMM) or human serum albumin alone (HSA-IMM), and treatment with gonadotropins (GTH) or saline. The GTH treatment was 2,000 IU PMSG + 1,000 iu hCG, and control heifers were given saline (s.c.). Immunization against GnRH (GnRH-IMM) caused cessation of ovulation (progesterone < 1 ng/mL plasma for 3 wk) and GTH treatment began within 3 to 5 wk after heifers became anovulatory. Heifers immunized against HSA (HSA-IMM) were treated between d 9 and 11 of the estrous cycle. All heifers were given PGF (i.m.) on d 2 (d 0 = start of GTH treatment) and 2,000 IU of hCG (i.m.) on d 4. Ovaries of heifers were evaluated by ultrasound. Blood was sampled daily from d 0 to d 8 and every second day through d 24 to quantify progesterone and estradiol. Concentrations of estradiol increased with time after GTH but the response was reduced by GnRH-IMM (IMM x GTH x day, P < .01). Number of follicles > or = 6 mm (d 0 to 14) was increased after GTH (GTH x day, P < .001) but was decreased (P < .03) by GnRH-IMM. Immunization against GnRH reduced total follicles > or = 6 mm on d 4 in heifers on both saline and GTH treatments. Treatment with gonadotropins increased the number of follicles > or = 16 mm on d 14 compared with control heifers. The number of CL was increased by GTH (GTH x day, P < .001) but the response was reduced (P < .03) in GnRH-IMM heifers compared with HSA-IMM heifers. We conclude that treatment with gonadotropins will induce follicular growth and ovulation in heifers that are anovulatory due to immunization against GnRH. PMID- 8726743 TI - Acute and chronic changes in luteinizing hormone secretion and postpartum interval to estrus in first-calf suckled beef cows exposed continuously or intermittently to mature bulls. AB - The objective of this study was to determine whether patterns of LH secretion are acutely or chronically affected by the presence of mature bulls in postpartum first-calf suckled beef cows exposed to bulls either continuously or intermittently beginning on d 30 after birth. Crossbred cows were assigned randomly to be either continuously exposed to (BE; n = 20) or isolated from bulls (NE; n = 32) at calving, or exposed continuously (NEBE; n = 10) or intermittently (BEI; n = 21) to bulls beginning on d 30. The BEI cows were exposed to bulls for 2 h every 3rd d for 18 d. Ten cows from the NEBE, BEI, and NE treatments only were fitted with indwelling jugular catheters. Intensive blood sampling of NEBE and BEI cows began within 45 min after they were placed with bulls for 2 h on d 30, and for BEI cows on each d after 2 h exposure. Samples were collected at 15 min intervals for 6 h beginning on d 30 for NEBE, BEI, and NE cows; sampling continued at 3-d intervals until d 48. Samples were assayed for LH by RIA. Cows were observed twice daily (am:pm) for estrus. More (P = .07) BE and NEBE (75%) cows showed estrus by the end of the study than BEI and NE cows (48%). Interval to estrus was longer (P < .05) in BEI and NE cows (95.6 +/- 6.1 d) than in BE and NEBE cows (75.9 +/- 6.1 d). Baseline LH and amplitude and interpeak interval of LH peaks during the first 6 h after 2 h of bull exposure did not differ (P > .10) among treatments on d 30 after birth. However, mean LH and LH pulse frequency were higher (P = .06) for NEBE and BEI cows than in NE cows. Baseline LH and amplitude and duration of LH peaks did not differ (P > .10) over the seven sampling periods among NEBE, BEI, or NE cows. Mean LH and LH pulse frequency were higher (P < .05) in NEBE and BEI cows than in NE cows on each sampling period. Exposing first-calf suckled beef cows to bulls on d 30 after birth increased mean LH concentrations by increasing pulse frequency within a short period after a 2-h exposure. Thereafter, mean LH concentrations were higher in cows that were either continuously or intermittently exposed to bulls. Although mean LH and LH pulse frequency in NEBE and BEI cows were similar, intervals to estrus after bull exposure differed between treatments. The mechanism whereby bulls alter postpartum interval to estrus seems to involve other factors that may be related to but not directly linked with LH secretion. PMID- 8726744 TI - Effect of maturity on digestion kinetics of water-soluble and water-insoluble fractions of alfalfa and brome hay. AB - Alfalfa and bromegrass, each harvested at five different stages of maturity, were separated into water-insoluble and -soluble fractions. The NDF concentrations ranged from 19 to 43% for alfalfa and from 42 to 58% for brome. The rates of digestion, by mixed ruminal microflora, of the unfractionated forage and of the water-insoluble and -soluble fractions were measured in vitro using pressure sensors to monitor gas production. Both forages showed the expected decline in fiber digestibility with increasing maturity. A dual-pool logistic model gave pool sizes, specific rates, and a single lag time for both the faster- and slower digesting fractions. The main difference between alfalfa and brome was in the soluble pool. This pool produced approximately 40% of the total gas in alfalfa, 25% in brome. The specific digestion rates of the brome soluble pool were approximately 50% higher than those for alfalfa. Net VFA production showed a somewhat higher acetate: propionate ratio for brome (3.2) compared with alfalfa (2.2), but there was little change with increasing maturity within a given forage. Gas production curves for the unfractionated forage showed a 0 to 10% positive deviation from curves created by adding data from separate digestion of the insoluble and soluble forage fractions. Gas measurements offer a promising approach to the study of the water-soluble extracts of forages and the interaction of the soluble- and insoluble-fractions during fermentation. PMID- 8726745 TI - Supplemental cracked corn or wheat bran for steers grazing endophyte-free fescue pasture: effects on live weight gain, nutrient quality, forage intake, particulate and fluid kinetics, ruminal fermentation, and digestion. AB - Two experiments were conducted with beef steers (Exp. 1, average BW of 580 kg; Exp. 2, average BW of 247 kg) to evaluate the use of no supplements (CON) or daily supplementation with (OM basis) .34% of BW of cracked corn (CORN), .34% of BW of wheat bran (WBBW), or .48% of BW of wheat bran (WBISO; calculated to be isocaloric to CORN) on digestive responses (Exp. 1) and live weight gain (Exp. 2). In Exp. 1, type of supplement did not affect (P > .10) the dietary fiber or N constituents, but in vitro OM disappearance of the forage differed (P < .10) with supplementation and type of supplement fed. Supplemented steers consumed less (P < .05) forage and total OM. Particulate passage, fluid passage, and ruminal pH were not affected (P > .10) by supplementation. Ruminal NH3 N concentration showed (P < .05) a treatment x sampling time interaction and, in general, WBBW and WBISO steers had greater ruminal NH3 N than CORN and CON steers. Total VFA concentrations and molar proportions of propionate were lower (P < .10) in CON steers than in supplemented steers; no differences were noted (P > .10) among supplemented steers. Molar proportions of acetate were lower (P = .01) in supplemented steers than in CON steers and were greater (P = .03) in WBBW steers than in WBISO steers. Butyrate molar proportions were lower (P < .05) in CON steers than in supplemented steers and differed (P < .10) with type and quantity of supplement supplied. In situ forage NDF disappearance at 6, 9, and 24 h after feeding and rate of disappearance were greater (P < .05) in CON steers than in supplemented steers. In Exp. 2, CON steers weighed less (P = .01) than supplemented steers, CORN steers weighed more (P = .08) than wheat bran supplemented steers, and WBISO steers weighed more (P = .02) than WBBW steers; ADG for 90 d followed a similar response. Results suggest that supplementation of wheat bran rather than corn did not seem to stop the reduction in forage intake or OM digestion associated with corn supplementation. PMID- 8726746 TI - Application of nitrogen from swine lagoon effluent to bermudagrass pastures: seasonal changes in forage nitrogenous constituents and effects of energy and escape protein supplementation on beef cattle performance. AB - A 2-yr study was conducted to evaluate 1) the effects of nitrogen level from swine lagoon effluent on forage composition and animal performance and 2) the effects of supplemental escape protein (EP) on performance by steers grazing pastures fertilized with swine lagoon effluent. Bermudagrass (Cynodon dactylon) pastures were fertilized via sprinkler irrigation with either 448 or 896 kg/ha of N from swine lagoon effluent (two pasture replications/treatment). Within each pasture, four supplement treatments were evaluated using electronic Calan gates (8 steers.pasture-1.yr-1). The treatments were 1) negative control (pasture only); 2) energy control, which supplied 79.9 g of EP.hd-1.d-1; 3) 159 g of EP.hd 1.d-1, and 4) 239 g of EP.hd-1.d-1. Gains were increased (P < .05) an average of .15 kg/d by supplementation, with no differences in gain among supplements. Forage samples representative of that grazed by steers (CONSUM) and representative of all available forage (AVAIL) were obtained at 14-d intervals. Total CP content of CONSUM and AVAIL samples were slightly higher (P < .20 and P < .15, respectively) from pastures fertilized with 896 compared with 448 kg/ha of N (20.8 vs 20.0% for CONSUM and 15.2 vs 14.2% for AVAIL). Concentrations of nonprotein N and soluble true protein (% of CP) in both AVAIL and CONSUM samples were higher (P < .06) from pastures fertilized with the higher N level. Total N uptake by the plant, based on CP content and animal grazing days/ha, was essentially the same for both N treatments. PMID- 8726747 TI - Preservation of ruminal microorganisms for in vitro determination of ruminal protein degradation. AB - Ruminal microorganisms, preserved either lyophilized or frozen, were compared with freshly strained ruminal fluid for proteolytic activity and as inoculum source for determination of ruminal protein degradation rates by the inhibitor in vitro method. Dialysis and glycerol addition had no effect on the proteolytic activity of preserved microorganisms. Net release of NH3 and total amino acids from protein using the fluid plus particle-associated microorganisms was higher than that found using the fluid-associated microorganisms alone. Method of inoculum preservation altered total proteolytic activity, but harvesting bacteria using centrifugal force greater than 5,000 x g did not increase proteolytic activity of the pellet. The proposed method for harvesting and preserving microorganisms consisted of centrifuging strained ruminal fluid at 5,000 x g (30 min at 4 degrees C), stirring the pellet in a 50:50 (vol/vol) solution of glycerol-McDougall's buffer for 15 min, and then storing at -20 degrees C. Protein degradation rates in incubations with preserved microorganisms were four to eight times slower than when using fresh ruminal fluid; however, feed proteins were ranked similarly for degradation rate. Preincubating the preserved microorganisms reduced blank concentrations of NH3 and total amino acid and increased protein degradative activity of the preserved inoculum. Degradation rates with preincubated, preserved inocula were similar to those obtained using fresh ruminal fluid. These results indicated that mixed ruminal microorganisms can be preserved by freezing and, after a preincubation period of 6 h, used as the inoculum source for in vitro estimation of ruminal protein degradation. PMID- 8726748 TI - Effects of alfalfa root saponins on digestive function in sheep. AB - The effects of alfalfa saponins on ruminal pH and VFA concentrations, protozoal numbers, forestomach motility, and digesta flow from the rumen were examined in this experiment. In a preliminary study, either 800 or 1600 mg/kg of saponins per kg BW were administered intraruminally in a single dose to one of two 60-kg wethers. Ruminal contractions were suppressed within 15 min. The study was terminated when one wether didn't recover. Saponins were then administered intraruminally in two equal doses daily into four ruminally and duodenally cannulated wethers (60 +/- 1 kg) in amounts equivalent to 0, 200, 400, and 800 mg/kg BW daily in a 4 x 4 Latin Square arrangement. Saponins increased VFA concentrations (P = .001) and lowered ruminal pH (P = .001) after 2 d of administration. On d 2 and d 14, saponins decreased (P < .01) protozoal populations. On d 11, pressure changes during ruminal contractions increased by 79% during resting (P = .06), 202% during feeding (P = .05), and 53% during an 8 h period (P = .08) as the level of saponins increased from 0 to 800 mg/kg BW. Administered saponins increased duodenal flows of OM and total N by 19 and 26%, respectively (P = .04 and .01). Apparent total tract N digestibility was reduced (P = .02) by 5% at the highest saponin dosage. We conclude that alfalfa may contain enough saponins to adversely impact ruminoreticular motility. Saponins also caused decreased ruminal protozoal populations, increased flow of total duodenal N, and reduced ruminal and total tract apparent digestibilities. PMID- 8726749 TI - Influence of sampling time and diet on amino acid composition of protozoal and bacterial fractions from bovine ruminal contents. AB - Four ruminally cannulated cows were used to compare amino acid (AA) composition of protozoal and bacterial fractions as affected by sampling time and diet. Cows were given once a day restricted feed (80% of ad libitum intake) of 7 kg DM with two successive diets. Diet HB was 65% Cocksfoot hay and 35% pelleted ground barley, and Diet H was 100% Cocksfoot hay. Samples of whole ruminal contents were taken 2, 5, 8, 11, and 23 h after feeding for Diet HB and 2 h after feeding for Diet H to isolate the liquid-associated protozoa and bacteria (LAP, LAB) and particle-associated bacteria (PAB). At each sampling time, the AA compositions of the different microbial populations were determined. The AA profiles of the LAP were different from those of the bacteria for 13 AA out of 17 studied. Differences between AA compositions of LAB and PAB were also observed for 10 AA out of 17 studied. Irrespective of the microbial population, AA composition did not vary with sampling time after feeding diet HB (P > .05; except for arginine, glutamate, and glycine). The AA contents of none of the three microbial populations were affected (P > .05) by the diet except for leucine and glutamate (P < .01). The differences in AA profiles between LAP and bacteria and between LAB and PAB confirm the importance of the representativeness of the microbial reference sample for correctly estimating microbial AA flow into the small intestine. PMID- 8726750 TI - Prospects for expanded mohair and cashmere production and processing in the United States of America. AB - Mohair from Angora goats has been produced in the United States since the introduction of these animals from Turkey in 1849. Cashmere on Texas meat goats was reported in 1973, but domestic interest in commercial production did not occur until the mid-1980s. Since 1982, the average prices of U.S.-produced mohair and cashmere (de-haired) have ranged from $1.81 to $9.48/kg and approximately $55 to $200/kg, respectively. However, return to producers from mohair has been relatively constant, averaging $10.21/kg, due to the federal incentive program. Because this program is scheduled to terminate with final payment in 1996, the future of mohair profitability is questionable. Prospects for expanded mohair and cashmere production and processing in the United States are influenced by numerous interacting factors and potential constraints. These include the prospect that the goat and textile industries may no longer be profitable in the absence of clear government policies. Although selection may have slightly increased fiber production by Angoras (long term) and domestic meat goats (short term), availability of genetic resources may prove to be a constraint to increased fiber production by cashmere goats and improved meat production by both types of goat. Land resources are plentiful unless new government policies prohibit goats from vast tracts of rangeland and forest because of environmental concerns. Future demand is an unknown, but with increasing world population and affluence, prospects for long-term improved demand for luxury fibers seem good. Competition from foreign cashmere growers is expected, whereas, in the short term, mohair production overseas is declining. However, increased processing of cashmere in its country of origin is expected to result in shortages of raw materials for European and U.S. processors. The amount of scouring, worsted, and woolen equipment in the United States is adequate to accommodate major increases in domestic processing of goat fibers. However, the absence of specific processing knowledge and skills may be constraints. Similarly, the absence of acceptable small-scale dehairing equipment for cashmere will limit cashmere processing on a cottage industry scale. Purely practical considerations such as the effects of predation and cost of fencing could become major constraints to expanding the goat fiber industry. Likewise the success (or lack thereof) of industry promotion of fiber and goat meat could be an overriding factor. To emerge from the uncertainty of erratic raw material prices and to better control profitability, domestic goat-fiber producers are encouraged to consider innovative, cooperative, retained ownership business ventures that will permit them to profit-share up to the retail level. PMID- 8726751 TI - Status and prospects of the dairy goat industry in the United States. AB - Among the major classes of U.S. livestock, dairy goats have yet to achieve USDA statistical reporting of their numbers, amounts of milk produced and processed, and cheese and other products marketed. However, the USDA has published buck proofs of approximately 16,000 does annually from Dairy Herd Improvement Association (DHIA) records of the Alpine, LaMancha, Nubian, Oberhasli, Saanen and Toggenburg breeds, thereby encouraging genetic progress. This represents a 1% participation in DHIA of the estimated 1.5 million U.S. dairy goats. Annual breed registrations are led by Nubians (11,000), and the leading states in descending order are California, Texas, Ohio, New York, and Pennsylvania. Breed average milk yields range from 960 kg of milk for Saanen to 726 kg of milk for Oberhasli. Average milk contents range from 4.5% fat and 3.69% protein for Nubian to 3.3% fat and 2.98% protein for Toggenburg. Leading lactation records are 3,023 kg of milk (Toggenburg) and 174 kg of fat (Nubian). Total annual registrations are 45,000+ animals by 16,000+ member breeders. Estimated total U.S. goat milk commercial production is 24,000+ t, with half going into commercial farm goat cheese production of 640+ t. Recent years have seen significantly increased numbers of dairy goat research projects and publications from Oklahoma, Texas, California, Georgia, Alabama, Florida, Louisiana, New York, Connecticut, Delaware, and Massachusetts. Furthermore, annual national and international symposia, annual national goat cheese judging competitions and workshops, an active national goat research foundation, representation on the National Interstate Milk Shippers Committee and Mastitis Council, and formation of a national association and council for the development and promotion of dairy goat products indicate an evolution from former emphasis on purebred breed development to a focus on market development. The conclusion is that dairy goats are emerging as a necessary and recognized U.S. industry. PMID- 8726752 TI - Preclinical evaluation of pharmacotherapies for treatment of cocaine and opioid abuse using drug self-administration procedures. AB - Drug abuse is a major public health problem, and the relationship between intravenous drug abuse and AIDS underscores the need for more effective treatment medications. Animal models of drug self-administration are useful to systematically evaluate new treatment medications and predict clinical efficacy. This review summarizes the status of preclinical evaluations of medications for treatment of cocaine and opiate abuse. The basic drug self-administration methodology and the rationale for experimental designs and outcome criteria are described. Studies of the effects of dopamine or opioid receptor agonists and antagonists as well as medications used clinically for other indications on drug self-administration are critically examined. Where possible, the degree of concordance between clinical and preclinical studies of drug abuse treatment medications is discussed. We conclude that drug self-administration models are valuable for preclinical assessment of medication efficacy, and we recommend some strategies to further improve evaluation procedures. The discovery of more effective medications for substance abuse treatment should be facilitated by recent advances in behavioral science, pharmacology, neurobiology and medicinal chemistry. PMID- 8726753 TI - Chronic administration of serotonergic antidepressants attenuates the subjective effects of LSD in humans. AB - This study investigates the possible interactions of antidepressant agents and hallucinogens in humans through structured interviews using a standardized questionnaire. Volunteer subjects recruited through announcements placed on the Internet or other sources were asked to describe the somatic, hallucinatory, and psychological effects of self-administered LSD prior to and during chronic administration of an antidepressant. Twenty-eight out of 32 subjects (88%) who had taken an antidepressant with inhibitory effects on serotonin (5-HT) reuptake (fluoxetine, paroxetine, sertraline, trazodone) for over 3 weeks had a subjective decrease or virtual elimination of their responses to LSD. An additional subject who had taken fluoxetine for only 1 week had an increased response to LSD. These data are in contrast to our previous study that reported increased responses to LSD during chronic administration of tricyclic antidepressants or lithium. Possible mechanisms of action for the effects from serotonergic antidepressants involve 5-HT2 and 5-HT1A receptors, changes in extracellular brain serotonin concentrations, and changes in brain catecholamine systems. PMID- 8726754 TI - Fluoxetine-induced sleep disturbance in depressed patients. AB - Abnormal polysomnographic (PSG) features, most notably increased electromyographic (EMG) tone and eye movements during non-REM sleep have been observed during sleep in fluoxetine-treated depressed patients. However, the relationship between these PSG features and sleep disruption is unclear. Nine depressed patients treated with 10 to 80 mg of fluoxetine and six unmedicated, depressed patients were studied polysomnographically on two consecutive nights during which sleep parameters, transient arousals, and eye movements were measured. The fluoxetine group experienced a lower-average sleep efficiency index (SEI) and significantly more eye movements and arousals during non-REM sleep than the control group. Eye movement and arousal counts were significantly correlated. In addition, clinically significant periodic limb movement disorder (PLMD) was observed in 44% of the fluoxetine-treated group versus none of the control group. We conclude that a higher incidence of PLMD and frequent transient arousals associated with eye movements may be responsible in part for the complaint of insomnia made by patients treated with fluoxetine. PMID- 8726755 TI - Biochemical adaptations in the mesolimbic dopamine system in response to repeated stress. AB - We have demonstrated previously that chronic administration of morphine, cocaine, or ethanol produces some common biochemical adaptations in the ventral tegmental area (VTA) and nucleus accumbens (NAc), components of the mesolimbic dopamine system implicated in the reinforcing and locomotor activating properties of these drugs of abuse. Because this neural pathway is also regulated by stress, and because stress has been shown to influence an animal's behavioral responses to drugs of abuse, it was of interest to determine whether repeated exposure to stress results in similar biochemical adaptations. By use of immunoblot analysis, we show here that a course of chronic "unpredictable" stress, like chronic drug exposure, increased levels of immunoreactivity of tyrosine hydroxylase and glial fibrillary acidic protein and decreased levels of immunoreactivity of neurofilament proteins in the VTA. Chronic unpredictable stress also increased levels of cyclic AMP-dependent protein kinase activity and decreased levels of immunoreactivity of the G protein subunit, Gi alpha, in the NAc. These effects required long-term exposure to stress and were in most cases not seen in the substantia nigra and caudate-putamen, components of the nigrostriatal dopamine system studied for comparison. The biochemical effects of chronic stress in the VTA and NAc differed among three strains of rat studied. Fischer 344 rats were the most responsive in that they exhibited all of the aforementioned adaptations, whereas Lewis rats were the least responsive in that they exhibited none of these adaptations; Sprague-Dawley rats exhibited an intermediate number of responses. Taken together, the results of the present study demonstrate that chronic exposure to stress results in biochemical adaptations in the mesolimbic dopamine system that resemble the chronic actions of several drugs of abuse. These adaptations could contribute to the convergent behavioral effects induced by treatments that are mediated via the VTA-NAc pathway. PMID- 8726756 TI - Mycobacteria and glutaraldehyde: is high-level disinfection of endoscopes possible? AB - BACKGROUND: High-level disinfection of endoscopes has traditionally been undertaken by manual or automatic scope cleaning plus a 10 to 20 minute soak in 2% alkaline glutaraldehyde. Mycobacteria species are less sensitive to glutaraldehyde, and a 45-minute instrument soak has recently been recommended by the manufacturer. Because of concerns over endoscope damage, need for more endoscopes, and perception that the current cleaning method is adequate, we prospectively studied mycobacteria-contaminated endoscopes at various stages of the cleaning process. METHODS: All work was done under a laminar flow hood in a microbiology laboratory. Five gastrointestinal scopes were contaminated with 10(8) colony forming units per milliliter (CFU/mL) of Mycobacterium chelonei, an atypical mycobacterium similar in chemical resistance to Mycobacterium tuberculosis but with less infectious potential. Cultures of the sheath, biopsy channel, and elevator channel were taken at baseline, after manual cleaning, and after 10, 20, and 45 minutes to glutaraldehyde soak both before and after alcohol rinse. RESULTS: Manual cleaning resulted in a mean of 4.7 log10 reduction in viable mycobacterial colonies. Qualitative studies of the external endoscope surface as well as the air-water valve showed no detectable organisms after a 10 minute exposure to alkaline glutaraldehyde. Conventional quantitative culture techniques of the channels demonstrated one endoscope out of five with consistent growth after a 10-minute exposure to glutaraldehyde. Following alcohol treatment, there was no significant colony growth. In contrast, a quantitative membrane filter system showed the presence of at least one mycobacterial colony in four out of five scopes after a 45-minute glutaraldehyde exposure. CONCLUSIONS: Additional studies utilizing a standardized mycobacterial species, inoculum size, and suspension characteristics are recommended to delineate adequate duration of disinfectant exposure time. PMID- 8726757 TI - Quality improvement in gastrointestinal endoscopy: microbiologic surveillance of disinfection. AB - BACKGROUND: Cleaning and disinfection procedures play an essential role in the prevention of infection transmission in gastrointestinal endoscopy. In spite of published detailed guidelines, several variants and weak points still exist. METHODS: Cleaning-disinfection procedures were carried out according to "Working Party, Sydney 1990." A microbiologic surveillance protocol tested the contamination of endoscopes and of automatic washing machines. To assess and improve the efficacy of disinfection, we adopted a quality assurance program. RESULTS: During a 2-year follow-up, the outside surfaces of gastroscopes were contaminated in 60.5% and channels in 41.3%; the outside areas of colonoscopes were contaminated in 62.3% and channels in 40.3%. Isolated bacteria were gram negative organisms, particularly Pseudomonas species, and gram-positive organisms, mostly Staphylococcus species. The water reservoirs of automatic washing machines were frequently contaminated by Pseudomonas aeruginosa. The disinfection of washing machines and alcohol rinsing of endoscopes after standard procedures significantly reduced the bacterial contamination. CONCLUSIONS: The microbiologic surveillance pointed out the main weak points that could be improved by the adoption of corrective interventions. Quality assurance is a feasible method to assess the efficacy of cleaning-disinfection, and its wide application would improve quality of care. PMID- 8726758 TI - Colonic mucosal abnormalities associated with oral sodium phosphate solution. AB - BACKGROUND: Oral sodium phosphate (NaP) is increasingly used as a colonic cleansing agent for colonoscopy. It has been shown to be efficacious, well tolerated, and safe. Mucosal abnormalities associated with NaP have recently been described. We carried out this controlled study to assess whether bowel cleansing preparations commonly used in colonoscopy are associated with colonic mucosal changes that may mimic inflammatory bowel disease (IBD). METHOD: All patients undergoing colonoscopy from January 1994 to June 1994 were considered for the study. Patients with history or symptoms suggestive of IBD were excluded. Patients were randomized to receive polyethylene glycol-based lavage (PEG-ELS) or NaP solution as their bowel cleansing preparation. Two gastroenterologists performing the colonoscopies were blinded to the type of preparation. Any mucosal abnormalities were noted and photographic documentation and biopsy specimens were taken. RESULTS: Ninety-seven patients were studied, 44 receiving PEG-ELS and 53 receiving NaP. Both groups were similar with regard to sex, age, and indication for colonoscopy. Fourteen patients were found to have nonspecific aphthoid-like erosions similar in appearance to Crohn's disease. These lesions, however, were not friable and biopsy results were not compatible with IBD. This mucosal abnormality was found in 13 patients who received NaP (24.5%) and only 1 patient who received PEG-ELS (2.3%). CONCLUSION: Nonspecific aphthoid-like mucosal lesions occur frequently in patients who received NaP for colonoscopy preparation. These lesions are endoscopically similar to those seen in Crohn's disease. Because of the potential for misinterpretation of these lesions, we do not recommend the use of NaP as a colonic cleansing preparation for patients with chronic diarrhea or in whom the diagnosis of IBD is suspected. PMID- 8726759 TI - Effect of oral sodium phosphate colon preparation on serum electrolytes in patients with normal serum creatinine. AB - BACKGROUND: Oral sodium phosphate is commonly used for colon preparation prior to colonoscopy. Several studies have demonstrated significant changes in serum electrolytes after sodium phosphate preparation, but failed to stratify patients with regard to renal function. The purpose of this study was to determine if electrolyte alterations are observed after sodium phosphate in patients with normal levels of serum creatinine. METHODS: Thirty-two patients scheduled for elective colonoscopy who had a serum creatinine level of less than 1.5 mg/dL were enrolled. All patients had a panel of laboratory studies prior to and immediately after receiving oral sodium phosphate colon preparation. RESULTS: Significant increases in serum phosphate and sodium and decreases in serum calcium and potassium were observed. Twenty-eight percent of patients had serum phosphate levels greater than 8.0 mg/dL, and 6% of patients had serum calcium levels less than 8.0 mg/dL after sodium phosphate colon preparation. There were no clinically apparent adverse events. CONCLUSIONS: Significant changes in serum electrolyte levels occur after sodium phosphate colon preparation in patients with normal serum creatinine levels. Sodium phosphate could exacerbate pre-existing hypocalcemia or hypokalemia in patients with apparently normal renal function. Therefore, it may be prudent to evaluate serum electrolytes prior to administration of sodium phosphate. PMID- 8726761 TI - Benign esophageal strictures in children and adolescents: etiology, clinical profile, and results of endoscopic dilation. AB - BACKGROUND: The problem of dysphagia in children and adolescents differs from that in adults, and therefore requires special consideration. METHODS: Forty-one consecutive children and adolescents 16 years of age or younger (mean, 7.2 years), with benign esophageal strictures were evaluated in a prospective manner over a 7-year period. The most frequent causes of esophageal strictures were caustic ingestion and complications of endoscopic sclerotherapy of esophageal varices. Dilation was done on a weekly basis using bougies and was considered adequate if the esophageal lumen could be dilated to 15 mm diameter (11 mm in children less than 5 years old) with complete relief of dysphagia. RESULTS: Of the 30 patients who could be adequately followed after initial dilation, 16 had corrosive strictures and 14 had strictures due to other causes. Patients with corrosive strictures required a significantly higher number of sessions for adequate initial dilation (7.8 +/- 2.5 sessions vs 1.86 +/- 0.48 sessions; p < 0.01). Patients with corrosive strictures had a higher number of mean symptomatic recurrences per patient month as compared to the noncorrosive stricture group (0.15 +/- 0.01 vs 0.087 +/- 0.03, p < 0.01). Six esophageal perforations occurred during a total of 327 dilation sessions (1.8%); there was one fatality. CONCLUSIONS: From our experience, we conclude that benign esophageal strictures in young patients can be treated effectively and with acceptable safety by means of endoscopic dilation. PMID- 8726760 TI - Natural history of bleeding peptic ulcers with a tightly adherent blood clot: a prospective observation. AB - BACKGROUND: The natural history of a bleeding peptic ulcer with a tightly adherent blood clot remains uncertain. Controversy exists concerning removal of such blood clots at the bleeding ulcer base. This article presents the natural history of a bleeding peptic ulcer with a tightly adherent clot and defines the characteristics of those requiring aggressive management. METHODS: Clinical parameters were analyzed to determine the independent predictors of rebleeding in these patients. One hundred one patients with bleeding peptic ulcers and tightly adherent blood clots were enrolled during a period of 12 months. RESULTS: Twenty five patients (25%) rebled within 1 month. With a multivariate analysis, we found comorbid illness (odds ratio, 3.41), shock (odds ratio, 3.65), and initial hemoglobin at or below 10 gm/dL (odds ratio, 2.99) to be independent predictors of rebleeding. CONCLUSIONS: Most patients with a tightly adherent clot in an ulcer have an uneventful course. However, endoscopic therapy may prove to be beneficial in the subset of patients with independent predictors of rebleeding. PMID- 8726763 TI - Treatment of esophageal obstruction from mediastinal compressive tumor with covered, self-expanding metallic Z-stents. AB - BACKGROUND: Mediastinal malignancies may involve the esophagus, leading to esophageal stenosis and dysphagia. Rigid and self-expanding esophageal stents have been used for effective palliation, but their use in extrinsic, compressive lesions is controversial. METHODS: A retrospective review of self-expanding Gianturco-Rosch Z-stents that were successfully placed in 13 patients with malignant esophageal obstruction due to extrinsic lesions. RESULTS: All patients had an improvement in dysphagia of at least two dysphagia grades. The mean dysphagia grade fell from 3.15 to 0.62. Mean survival was 2.2 months. Early (within 48 hours) procedure-related complications occurred in 4 of 13 patients and consisted of minor, transient chest pain that resolved within 6 hours (3 patients) and endoscopic stent dislodgment into the stomach (1 patient). Late complications (> 48 hours) occurred in 2 patients and consisted of a partial proximal stent migration and the development of a benign stricture proximal to the stent. There was no procedural or stent related mortality. CONCLUSIONS: Esophageal obstruction and malignant dysphagia from extrinsic, compressive mediastinal malignancies can be effectively and safely palliated with self expanding Gianturco- Rosch Z-stents. PMID- 8726762 TI - Plastic prosthesis versus expandable metal stents for palliation of inoperable esophageal thoracic carcinoma: a controlled prospective study. AB - BACKGROUND: Rapid palliation of malignant dysphagia is usually possible with endoscopic implantation of a plastic prosthesis, but this device has a high rate of complications. Recently, expandable metal stents, a new class of endoprosthesis, have become available and may reduce complication rates. METHODS: Thirty nine patients affected by esophageal thoracic cancer were randomly assigned to treatment with either a plastic stent (20 patients) or expandable metal stent (19 patients). The degree of palliation (expressed as dysphagia score) and incidence of complications (short- and long-term) were compared in both treatment groups. RESULTS: Technical success, as a percentage of successful intubation, was similar in both treatment groups (90% vs 94.7%, p = NS) and dysphagia scores improved significantly and similarly in both treatment groups. Nevertheless, complications and mortality related to implantation were significantly less frequent with metal stents than with plastic prostheses (complications: 0% vs 21%, p < 0.001; mortality: 0% vs. 15.8%, p < 0.001). Late complications included obstruction by food in both treatment groups (four cases with plastic stents vs four cases with metal stents), tube migration only with plastic prostheses (two cases) and tumor ingrowth only with metal stents (two cases). CONCLUSIONS: Expandable metal stents can be considered an effective and safer alternative to conventional plastic prostheses in the treatment of esophageal obstruction caused by inoperable cancer. PMID- 8726764 TI - Patient-controlled analgesia for conscious sedation during endoscopic retrograde cholangiopancreatography: a randomized controlled trial. AB - BACKGROUND: Adequate comfort is essential to patients undergoing invasive procedures. This study was designed to evaluate whether patient-controlled analgesia could improve sedation for ERCP. METHODS: Patients were randomized to receive standard sedation (n = 31) or patient-controlled analgesia (n = 31). The patients were blinded to the randomization. After the procedure the patient, physician, and nurse each rated their satisfaction with sedation using a verbal rating scale. RESULTS: There was no significant difference between the patient's mean satisfaction score for the conventional and patient-controlled analgesia groups (9.3 and 9.6, respectively, p = 0.5). The physicians rated sedation higher in the conventional group compared with the patient-controlled analgesia group (8.6 and 8.2, respectively, p = 0.02). Physicians and nurses' scores correlated (r = 0.53, p = 0.0001), but there was no correlation between scores reported by either physicians or nurses and the patients' scores (r = 0.2 and r = 0.05, respectively). Oxygen saturation less than 90% occurred for more than 1 minute in three patients who received standard sedation but in none who used patient controlled analgesia. CONCLUSION: This trial demonstrates that patient-controlled analgesia during ERCP is as effective as standard sedation with respect to patient satisfaction. Physicians and nurses, however, are not good proxies for assessing patient satisfaction. PMID- 8726765 TI - A novel technique for dilating difficult malignant biliary strictures during therapeutic ERCP. PMID- 8726766 TI - Complete tissue sampling of biliary strictures at ERCP using a new device. PMID- 8726767 TI - Endoscopic treatment of gastric angiodysplasia with elastic band ligation. PMID- 8726768 TI - Obstructive jaundice due to Coccidioides immitis. PMID- 8726769 TI - Gallstone ileus treated by extracorporeal shockwave lithotripsy. PMID- 8726770 TI - Crohn's esophagitis: a unique cause of esophageal polyposis. PMID- 8726771 TI - Hepatic portal venous gas after transgastric EUS-guided fine-needle aspiration of an accessory spleen. PMID- 8726772 TI - Small intestinal histoplasmosis: successful treatment with itraconazole in an immunocompetent host. PMID- 8726773 TI - Flexible gastrointestinal endoscopic reprocessing. PMID- 8726774 TI - Colonoscopy preps: what's best? PMID- 8726775 TI - Aphthoid lesions of the rectum. PMID- 8726777 TI - Preoperative preparation prior to colorectal surgery. PMID- 8726776 TI - Red ring sign versus aphthous ulcers of colonic mucosa? PMID- 8726778 TI - Dilation of esophageal strictures and subgroup analyses. PMID- 8726779 TI - Prospective screening of dyspeptic patients by Helicobacter pylori serology. PMID- 8726780 TI - The role of endoscopic ultrasonography in biliary tract disease, obstructive jaundice. PMID- 8726781 TI - Endoscopic manometry of the sphincter of Oddi: its usefulness for the diagnosis and treatment of benign papillary stenosis. PMID- 8726782 TI - Recovered memories in psychotherapy. PMID- 8726783 TI - A critical review of recovered memories in psychotherapy: Part I--Trauma and memory. AB - OBJECTIVE: The theoretical basis of the use of recovered memories in psychotherapy will be critically examined. METHOD: Literature will be reviewed on the nature of normal memory, and on the relationship of trauma to memory. RESULTS: Normal memories are surprisingly inaccurate. There is little evidence that normal memories can be repressed. There is no evidence that trauma makes repression more likely. CONCLUSIONS: "Recovery" of repressed memories is not consistent with the findings of empirical research. PMID- 8726784 TI - A critical review of recovered memories in psychotherapy: Part II--Trauma and therapy. AB - OBJECTIVE: The clinical implications of the use of recovered memories in psychotherapy will be examined. METHOD: The paper will review the relationship of trauma to psychopathology and discuss how traumatic histories might be dealt with in therapy. RESULTS: Trauma is a risk factor for psychopathology, but is only one of many etiological factors in mental disorders. The search for recovered memories in psychotherapy could present dangers for patients. CONCLUSIONS: The most reliable memories of trauma are those that have been present throughout the patient's life. PMID- 8726785 TI - The characteristics and intervention histories of incarcerated and conduct disordered youth. AB - OBJECTIVES: 1) To determine if incarcerated youth and conduct-disordered youth in treatment display similar risk factors associated with chronicity of antisocial behaviour; 2) to determine if incarcerated youth had experienced high numbers of interventions by many different agencies as has been reported for conduct disordered youth in treatment. METHOD: The files of 25 conduct-disordered youth in treatment and 25 incarcerated youth were examined to extract all relevant data. RESULTS: Incarcerated youth were similar to conduct-disordered youth with regard to the number of symptoms of disruptive behaviour disorders, age of onset of behavioural problems, and family backgrounds. Both groups had high numbers of interventions and agencies in their histories. Interventions began earlier for the conduct-disordered youth. CONCLUSION: The distinction between conduct disordered youth who are treated and youth who are incarcerated may be arbitrary and accidental. Given the similarity in the clinical characteristics, incarcerated youth should be offered help that is similar to that offered to conduct-disordered youth. Such help may lower the rate of recidivism. PMID- 8726786 TI - Suicide in old age: a tragedy of neglect. AB - OBJECTIVE: To investigate the incidence and treatment of depression in geriatric suicide. METHOD: All coroners' records, autopsy and police reports for suicide victims aged 65+ in Ontario (n = 543) over 3 years were examined. RESULTS: Over 80% of the elderly who committed suicide received no psychiatric referral. Of the sample, 87% were untreated while only 13% received antidepressants. Tricyclics, which are lethal in overdose, were the drugs of choice. None of the sample was treated with the safer specific serotonin reuptake inhibitors (SSRIs). Females were 3 times as likely to be treated as were males, and those seeing psychiatrists were 4 times more likely to be treated with antidepressants than those seeing general practitioners (GPs). The physically ill were rarely treated. CONCLUSIONS: These findings suggest that early geropsychiatric assessment and vigorous treatment could prevent many suicides in old age. PMID- 8726787 TI - Occurrence and effects of personality disorders in depression: are they the same in the old and young? AB - OBJECTIVES: To determine the frequency and effects of personality disorders on episodes of depression in elderly and young inpatients. Personality disorders are common and may affect the prognosis of Axis I disorders. METHODS: Clinical records of 89 elderly inpatients and a matched comparison group of 119 young inpatients were reviewed to confirm the diagnosis of a major depressive episode according to the DSM-III-R criteria. The frequency of personality disorder diagnoses in the 2 groups was determined. Within each group, severity, functioning, and treatment were compared between those with and without personality disorders. RESULTS: Personality disorders were diagnosed more frequently in the young (40.3%) than in the elderly (27%). Both rates were similar to previous reports. Cluster C disorders were the most common personality disorders found in the elderly, compared to cluster B disorders in the young. Personality disorder in the young was associated with longer episodes of depression (P = 0.035) and poorer family relations (P < 0.001); whereas in the elderly, personality disorder was associated with more severe episodes (P = 0.014). CONCLUSIONS: These findings suggest that the frequency and effects of personality disorders on the depressed patient may differ according to age. PMID- 8726788 TI - From evidence to conclusions in psychiatric research. AB - OBJECTIVE: To identify problems in the derivation of conclusions from evidence in psychiatry research. METHOD: The scientific model of falsificationism is described as determining the logical requirements for proving conclusions from research evidence. Common types of problematic conclusions are identified, and examples from the current research literature are given. RESULTS: Poorly formed conclusions are based on inadequate hypotheses, ambiguously phrased, blind to negative findings, fallacious in logic, or neglectful of alternative explanations. CONCLUSIONS: The risks of accepting poorly formed conclusions are premature closure of scientific investigations, poor basis for decisions in psychiatric practice, and provision of misinformation to patients. It is recommended that practitioners be attentive to this aspect of critically appraising research. PMID- 8726789 TI - A review of the psychobiology and pharmacotherapy of posttraumatic stress disorder. AB - OBJECTIVE: To review the literature on certain psychobiologic elements of posttraumatic stress disorder (PTSD) as they pertain to possible pharmacotherapeutic interventions. METHOD: The literature pertaining to the neuroanatomical, neurochemical, and cellular elements was reviewed. As well, both controlled and uncontrolled studies of pharmacotherapy in PTSD were analyzed. RESULTS: The literature suggests that the stress response triggers certain neuromodulators with subsequent psychoneurological restructuring; that various antidepressants have been demonstrated to be effective for treatment of criterion B symptoms; that, to date, a single antidepressant has been demonstrated to be effective in a controlled trial for criterion C symptoms; and that, to date, in controlled trials, antidepressants and a benzodiazepine have proved effective for criterion D symptoms. CONCLUSION: Currently, a comprehensive approach requires multimodel understanding and multimodal treatment. PMID- 8726791 TI - [Capgras syndrome: open perspectives for cognitive neuropsychology]. AB - OBJECTIVE: To examine the way in which certain concepts regarding the physiopathology of Capgras' syndrome (1) have been tested neuropsychologically. METHODS: Data consist of approximately 30 studies selected for their relevance to the cerebral stages of face processing in patients with schizophrenia, patients with Capgras' syndrome and normal subjects. RESULTS: Study of this work shows: a) that with respect to patients, authors have focused on the stage of treatment corresponding to the facial recognition phase per se; b) but that it is also possible to study the phase corresponding to knowledge and beliefs relative to individuals and to evaluate the existence of the cleavage proposed by numerous psychodynamicians. CONCLUSIONS: Views from the field of neuropsychology, like those from the field of psychodynamics, can therefore be tested. By offering a means of developing testable predictions in experimental protocols, cognitive neuropsychology methods will, in short, make it possible to reject erroneous concepts and demonstrate accurate ones. Limited here to the example of Capgras' syndrome, we advocate that the same methods be applied to Capgras' syndrome as to each symptom of schizophrenia. PMID- 8726790 TI - A double-blind, multicentre study of paroxetine and maprotiline in major depression. AB - OBJECTIVE: This study was performed to compare the clinical efficacy, side effects, and safety of paroxetine and maprotiline, the latter being the most frequently prescribed antidepressant in Switzerland. METHOD: Seventy-one patients (in and outpatients) with major depression were randomly allocated to treatment with paroxetine (20 to 40 mg daily) or with maprotiline (50 to 150 mg daily). Efficacy was measured by means of the Hamilton Psychiatric Rating Scale for Depression, the Montgomery-Asberg Depression Rating Scale, the Clinical Global Impression, and the Hopkins Symptom Checklist. RESULTS: The 2 components showed a similar efficacy. The adverse effect profile was comparable in the 2 treatment groups, although the findings showed a nonsignificant trend pointing in the direction of lower side effects with paroxetine. CONCLUSION: In the moderate dose regimens tested, the 2 components seemed to be of similar efficacy, with comparable profiles of side effects and safety. PMID- 8726792 TI - Re: The limits of small area analyses for health planning. PMID- 8726793 TI - Clozapine-induced neuroleptic malignant syndrome: clozapine monotherapy rechallenge in a case of previous NMS. PMID- 8726794 TI - The use of etiology in DSM-IV. PMID- 8726795 TI - Charles Bonnet syndrome: incidence and demographic and clinical features. PMID- 8726796 TI - Syndrome of "reverse" intermetamorphosis. PMID- 8726797 TI - Etorphine: a new opiate of abuse? PMID- 8726798 TI - Circadian cell kinetics in humans. Aspects related to cancer chemotherapy. AB - A prerequisite for the investigation of cell kinetics, and especially circadian cell kinetics, has been the development from the 1950s and onwards of several methods for studying kinetic parameters in different mammalian tissues. A large number of such studies have subsequently taken place in the rodent, mostly as non circadian experiments, but also a large number of studies have now documented on circadian proliferative rhythms in many different murine tissues. Results from cytokinetic studies in the human have also accumulated through the years. Of special interest has been the demonstration of temporal variations in rapidly proliferating tissues studies as the bone marrow and gut mucosa relative to cytotoxic anticancer therapy. Analyses of proliferation in human tumours have also indicated rhythms in malignant solid tumours. Thus, these studies have demonstrated that there exist rhythms in bone marrow and gut cytokinetics which increases the likelihood that certain times of day will be less toxic for the administration of cytotoxic drugs. Furthermore, optimizing anticancer therapy according to a circadian schedule may also increase the tumour cell death rat, due both to an indirect dose-escalating effect and a direct increased tumour effect. PMID- 8726799 TI - Importance of cell kinetics rhythmicity for the control of cell proliferation and carcinogenesis in rat liver (review). AB - The circadian control of cell Proliferation and Differentiation has been studied principally in rat liver. The comparison between the differentiation by hepatic enzymes and the division by the cell cycle under various experimental conditions (postnatal maturation, regeneration after partial hepatectomy, adrenalectomy, corticosterone treatments etc.) leads to the following conclusions: Under physiological conditions, proliferation and differentiation activities present a mutually exclusive relationship with a specific circadian rhythm. For both functions, the circadian variation of corticosterone plays the role of synchronizer, each evening (peak) it induces the synthesis of tissue specific enzymes in G0 cells and simultaneously inhibits the DNA synthesis in cycling cells. The same parameters have been studied during the different stages of hepatocarcinogenesis induced by Diethylnitrosamine (DEN). After initiation alone, (DEN for 2 weeks) circadian control is unchanged and precancerous cells are not able to reach malignancy. Promotion (DEN for 6 weeks) consists of disturbing the circadian synchronization to liberate the selective growth of initiated precancerous cells. This proliferation advantage favours the accumulation of chromosomal aberrations including those implicated in malignant transformation: i.e. activation of oncogenes or inhibition of antioncogenes. PMID- 8726800 TI - Chronotherapy for human solid tumors other than colorectal. AB - Available data regarding the application of chronobiological concepts to the medical treatment of various human solid tumors are reviewed. In the majority of reported trials, an improvement of therapeutic index has been observed with an increase in overall drug dose intensities, despite reduced overall toxicity and an excellent level of tumor response and survival. This assessment was obvious for ovarian, lung (non small cell), renal cell, pancreas, breast and upper aerodigestive tract carcinomas. PMID- 8726801 TI - Hepatic resections after chronotherapy: surgical aspects. AB - Hepatic resection has always been recommended for the treatment of metastatic disease in selected patients. Surgeons have had modified their approach and timing of surgery since the introduction of chemotherapy. The authors report their experiences since the introduction of chronochemotherapy. From 1990 up to 1994, 9 hepatic resections were performed for metastatic colorectal carcinoma. During the same period, 11 arterial accesses were implanted for diffused hepatic disease. Of the 9 resected patients, 5 are still alive with a mean follow-up of 17.7 months with no signs of recurrent disease. PMID- 8726802 TI - Clinical chronotolerance to anticancer drugs: relevance for dose-intensity. AB - There is increasing experimental and clinical data to suggest that the timing of chemotherapy may be important for both toxicity and response. The availability of programmable infusion pumps has made the clinical testing and application of circadian scheduling possible. This paper provides an overview regarding the current clinical data with some references to the experimental studies. Further progress in this area is dependent on a better understanding of the mechanisms involved and continued high quality clinical trials. PMID- 8726803 TI - Ambulatory chrono-chemotherapy by portable pumps: feasibility and compliance. Nursing aspects. AB - A study was undertaken among thirty seven advanced cancer patients, receiving chronochemotherapy by ambulatory programmable-in-time pumps. Drugs were infused through simple or double chamber venous, and/or arterial totally implantable side ports. The aim was to evaluate the treatment feasibility in an ambulatory mode, while appreciating the patient's physical and psychological tolerance and measuring the treatment's impact on the patient's daily life and family unit. The results of the study showed that, out of a total of 1613 days of treatment, only 27 returns to the hospital were required, which were due to minor incidents (mainly pipe leaks). No treatment was abandoned or interrupted by non-compliance and all patients maintained the ambulatory mode of treatment. Moreover patients cooperated fully with this mode of treatment with firm support from their relatives. The study emphasized the necessity of proper training for patients and good information about the delivery system, as a means of preventing the poor functioning of equipment and the ability to take promp action in order to maintain life functions and to confront potential side effects. PMID- 8726804 TI - Relevance of circadian cell kinetics in the timing of chemotherapy in animal tumors. AB - Circadian rhythmicity in cell division has been proved in the actively proliferating healthy tissues of rodents (cornea, G.I.-tract, bone marrow, gonads, epidermis) as well as in most spontaneous or transplanted tumors growing in solid or ascites fluid phases. This circadian division accounts in part for the circadian varying sensitivity of healthy target tissues to oncolytic agents (chronotoxicity/chronotolerance). Similarly, antitumor activity as gauged from analysis of impact on cell kinetics and/or tumor shrinkage or cure was also shown to depend upon the dose and doing time of anticancer drug administration. Variations in the timing of internal cell kinetics after initial drug administration were also shown to depend on the type of drug (alkylating agent, antimitotic, or antimetabolite agent). Interestingly, internal desynchronization may occur in both healthy and tumor tissues, it may allow the selection of given circadian stages for the administration of a second drug at the point of highest tumor sensitivity and healthy tissue resistance (time of least sensitivity). These observations have been utilized to test strategies in sequential drug scheduling aimed at improving the overall therapeutic index. Observations indicate that in some instances these approaches could be translated to human beings. PMID- 8726805 TI - Comparative psychological aspects of two different types of chemotherapeutic administration (chronotherapy vs. traditional chemotherapy) on quality of life of cancer patients at advanced stage. AB - In addition to qualitative information, specific quantitative psychiatrics tests (regarding anxiety and depression) and objective psychological tests specific to cancer populations were used to compare psychological variables in two groups of metastatic cancer patients treated with chemotherapy. The study also recorded 24 possible symptoms (somatic and psychic) allowing the self-evaluation of individual quality of life. The evaluation was performed at 7 different times (from before the beginning of treatment up to the 6th course). In comparison to traditional treatment, the results showed a better psychosocial adaptation for patients receiving chronoprogrammed administration of anticancer medication, with better social relations, less feelings of loss of independence, less anxiety, depression, and somatic discomfort. PMID- 8726806 TI - Stromal cell involvement in leukemogenesis and carcinogenesis. AB - There is increasing evidence that the supportive cells (stromal cells) in nearly all organs containing cellular self-renewal systems are involved in carcinogenesis. One body of evidence specific to irradiation leukemogenesis documents the role of irradiated murine stromal cells in the cell biologic changes associated with evolution of leukemia in cocultivated, nonirradiated stem cells. Stem cell phenotypic changes that have been documented include upregulation of cell surface c-fms, downregulation of growth requirement for obligatory growth factors, and the appearance of novel transcripts detected by differential display. A second body of evidence documents the potential role of stromal cells functioning as biologic tumor promoters through their release of reactive oxygen species (ROS), and production of altered adhesion molecules or growth factors during the chronic response to chemical or physical carcinogens. These molecular biologic mechanisms, potentially operative in stromal cells, can block apoptosis and induce DNA strand breaks in closely associated self-renewing stem cells. In an in vivo model of irradiation effects on lung stromal cells, we have irradiated the lungs of control C57BL/6J mice or other mice with orthotopic Lewis lung tumors and shown that TGF-beta release is increased following irradiation. The TGF-beta increase by irradiation may specifically be inhibited by administering an inhalation plasmid liposome mixture containing a transgene for human manganese superoxide dismutase prior to irradiation. An appreciation of the role of stromal cells in leukemogenesis and carcinogenesis may also be very relevant to the design of new therapeutic strategies for treatment of cancer, particularly since current strategies focus on eradication of stem cell transformants and do not rigorously address the persistence of surviving stromal cells. PMID- 8726807 TI - Induction of differentiation in embryonic stem cells by 26-kD membrane-bound tumor necrosis factor (TNF) and 17-kD free TNF. AB - The biological activity of 26-kD membrane-bound tumor necrosis factor (TNF)-a in embryonal development was examined in an in vitro system using embryonic stem (ES) cells. ES cells were seeded on NIH3T3 feeder cells transformed with mouse precursor TNF-a gene to express membrane-bound TNF-a on their cell surface. The proliferation of the ES cells was reduced and differentiation was accelerated. The same effects were also observed when 17-kD free TNF-a was added to the culture medium of the ES cells. Since free TNF-a is not present during embryogenesis, these results suggest that membrane-bound TNF-a may play an important role in embryonal development through cell-cell contact. PMID- 8726808 TI - Estradiol influences p53 expression in a human endometrial adenocarcinoma heterotransplanted into nude mice. AB - The influence of different estradiol concentrations on the expression of the p53 suppressor gene and on cell kinetics was examined by semiquantitative analysis of protein and bromodeoxyuridine labelling in a human endometrial adenocarcinoma grown in nude mice. We found that increasing the circulating estradiol increases (p = 0.001), and decreasing the hormone value decreases (p = 0.001) the expression of p53 in this tumor. The number of cells in the G1/G0 phase of the cell cycle was significantly higher (p = 0.03), and the number of cells in the G2/M phase was significantly lower (p = 0.01) in tumors grown in estradiol treated mice than in tumors obtained from the nontreated group. Changes in p53 expression may possibly be explained by either altered transcription activity of the gene or increased half-life of the protein. Our results suggest an important role of estradiol in the progression of estrogen receptor (ER) positive human endometrial adenocarcinomas. PMID- 8726809 TI - HIV-1 infectivity of human T cells in a human/murine chimeric fetal thymic organ culture system. AB - Human cord blood (HuCB) can colonize a murine fetal thymus organ culture (FTOC) and generate phenotypically immature (CD4+ CD8+) and mature (CD4+ CD8-; CD4- CD8+) T cells. We have used this model system to demonstrate that the human T cells that develop in this culture system can be infected with HIV-1. A cytopathic and non-cytopathic patient isolate of HIV-1 were used to infect FTOC established using C.B-17 or NOD/LtSz.scid/scid strain fetal thymic lobes colonized with HuCB. At 13-15 days after infection, FTOC were placed in co culture with human PHA-blasts. These co-cultures demonstrated the presence of replicating HIV-1. Few human CD45+ cells were detectable in the thymic lobes that were infected with HIV-1, while high numbers of human CD45+ T cells were present in the uninfected cultures. These results demonstrate the cytopathicity of HIV-1 on human T lymphocytes that have developed in a HuCB colonized FTOC system. PMID- 8726810 TI - Identification of neural crest derived cells within the cellular microenvironment of the human thymus employing a library of monoclonal antibodies raised against neuronal tissues. AB - The organization of optimal microenvironmental conditions within the developing thymus for lymphatic stem cell migration and their further maturation requires cellular and humoral participation of the neural crest. Recently, the immunophenotypical (IP) heterogeneity of lymphatic cells has become a scientific fact. Monoclonal antibodies (MoABs) produced against the various subpopulations of the reticulo-epithelial cells (RE) demonstrated their heterogeneity. We suggest that with a library of MoABs, raised against normal neuronal tissues, neural tumors, and a medulloblastoma cell line, including UJ13/A, UJ127.11, UJ167.11, UJ223.8, UJ308, J1153, A2B5, 215.D11, 275.G7, 282.1, antineurofilament (NF - med. m.w.), and anti-Thy-1 it is possible to recognize cells of neural crest origin within the postnatal thymic cellular microenvironment. Evidence has been collected concerning such connections between the nervous system and the thymus, such as the production of neuropeptides, oxytocin, and neurophysin by the thymus. Our immunohistochemical study was carried out on quick-frozen sections of human postnatal thymuses removed during open heart surgery, employing an indirect, alkaline phosphatase conjugated streptavidin-biotin technique. The employed MoABs reacted with the subcapsular (outer cortex) thymic nurse cells (TNCs) and with medullary RE cells, in close contact with already mature, immunocompetent T lymphocytes ready to leave the thymic microenvironment and enter the peripheral blood. The thymic medulla's strong immunoreactivity with A2B5, which binds to the GQ ganglioside, is typical for peptide secreting cells often migrated from the neural crest. A2B5+, Thy-1+ IP was demonstrated on the large TNCs. Cortical RE cells showed reactivity with UJ127.11, UJ223.8, and UJ308. Dense expression of neural crest antigens was detected in the Hassall's bodies (HBs) employing MoABs UJ223.8, UJ308, 215.D11, and 275.G7. These results suggest a neural crest origin for TNCs and for 20% to 30% of the cells of thymic microenvironment. The outer (peripheral) part of the HBs contained functionally very active RE cells. These RE cells also expressed antigens characteristic of the neural crest, detectable with MoABs UJ127.11, UJ223.8, UJ308, J1153, 215.D11, 275.G7 and A2B5. PMID- 8726811 TI - THNLA-1: a DNA-targeted bioreductive agent as chemosensitizer in vitro and in vivo. AB - THNLA-1 is a recently synthesized 2-nitroimidazole based, DNA-affinic bioreductive agent. It features a tetrahydroacridinic chromophore, which allows loose binding to DNA and therefore greater mobility along its backbone. THNLA-1 was proved to be a very good radiosensitizer and cytotoxin of hypoxic cells in vitro with an improved therapeutic index compared to the fully aromatic analog NLA-1. In this report we investigated the interaction of THNLA-1 with cis-DDP or L-PAM in the sensitive V79 and resistant OVCAR cells, using various schedule protocols. Also, the THNLA-1/cis-DDP interaction in balb/c mice has been investigated, using the EMT-6 mouse tumors. Isobologramic as well as fractional product concept analysis, clearly showed that synergistic interaction occurs between THNLA-1 and each chemotherapeutic agent, under hypoxic pretreatment conditions of the cells with THNLA-1 in vitro. The dose modification factor (DMF) values obtained in the resistant OVCAR-3 cells are similar to those obtained for the approximately 4 times more sensitive V79 cells. Therefore, the DMF value for e.g. L-PAM at 0.1 survival fraction, is approximately 2.76 when 15 microM THNLA-1 was used in OVCAR-3 cells, and approximately 2.50, when 10 microM THNLA-1 was used in V79. The supra-additive effect is dependent on the hypoxia-pretreatment time with THNLA-1, on THNLA-1 concentration and on the concentration of the chemotherapeutic drug. The limited in vivo study showed that THNLA-1, at doses significantly lower than its MTD, strongly potentiates the killing effect of cis DDP and that the optimum effect during the combination treatment was observed when THNLA-1 was administered i.p., 2.5-3.0 h before cis-DDP. Toxicity studies in balb/c mice (without tumors) showed that THNLA-1 is well tolerated up to at least 70 mg/kg for more than 40 days while no toxicity was observed with the combined drugs used in our experimental protocol. These results are promising for the potential clinical use of THNLA-1 as an adjuvant in chemotherapy. PMID- 8726812 TI - A role of transforming growth factor-beta 1 in the control of corneal neovascularization. AB - Transforming growth factor beta 1 (TGF-beta 1) has been implicated in both the stimulation of angiogenesis in vivo and in the inhibition of endothelial cell proliferation in vitro systems. In this study we present evidence showing that under certain experimental conditions TGF-beta 1 may inhibit neovascularization in vivo. TGF-beta 1 was incorporated into ethylene vinyl acetate copolymer (Elvax 40) matrixes which provide a valuable vehicle for the controlled and sustained delivery of bioactive peptides. The biological effectiveness of TGF-beta 1 sequestered in polymer matrices was assessed by measuring the inhibition of [3H] thymidine incorporation into the DNA of cultured mink lung epithelial cells. Neovascularization was induced in both corneas of albino rabbits by one deep seated limbal silk suture. Elvax 40 matrixes loaded with TGF-beta 1 (release rate, 1.66 ng/24 h) were implanted in rabbit corneal stroma. "Empty" polymers in the contralateral eye served as controls. Aliquots of aqueous fluid were withdrawn, and the presence activity of phagocytic cells was assessed by the production of superoxide anion (O2) which was measured by the superoxide dismutase-inhibitable reduction of ferricytochrome C. Polymer-enclosed TGF-beta 1 implanted in rabbit corneas significantly suppressed angiogenesis (2.65 +/- 0.4 mm compared to 3.05 +/- 0.3 mm in contralateral controls p < 0.05). Superoxide production in 100 microliters aliquots of aqueous fluid was 0.95 +/- 0.18 and 0.6 +/- 0.18 nmol O2/10 min in control eyes and in the eyes bearing sequestered TGF beta 1, respectively (p < 0.02). These results indicate that under the experimental conditions selected in this study, TGF-beta 1 significantly suppressed in vivo angiogenesis. PMID- 8726814 TI - Intraperitoneal administration of ascorbic acid delays the turnover of 3H labelled cortisol in the plasma of an ODS rat, but not in the Wistar rat. Evidence in support of the cardinal role of vitamin C in the progression of glucocorticoid synthesis. AB - The purpose of this investigation was to reproduce in rats the enhancing effect of vitamin C on adrenal glucocorticoid production, a phenomenon that has been repeatedly observed in our human experiments. Here, we tried to assess the impact of vitamin C pretreatment on the turnover of exogenous 3H-labelled cortisol tracer in rats. 500 mg ascobic acid in 15 ml isotonic liquid per rat was administered intraperitoneally into rats of the experimental group, and 15 ml physiological saline per rat was similarly introduced into rats of the control group. At various times after the loading of either vitamin C or saline, 3H labelled cortisol, 20 muc/0.5 ml saline/rat, was injected subcutaneously into rats of both the experimental and control groups. Thirty minutes later, all rats were sacrificed by exsanguination, and the radioactivity content was measured in the lipophilic fractions as well as in the purified corticosteroid compounds of blood, adrenal, testis and liver samples. To test the specificity of vitamin C action, experimental results from scurvy-prone ODS rats were compared with those of scurvy-resistant Wistar rats. Results obtained are as follows: a) the practice of vitamin C loading markedly delayed the turnover of 3H-cortisol in both the plasma lipophilic fraction and the plasma cortisol fraction, a finding which indicates that the above vitamin C pretreatment enhanced the release of endogenous glucocorticoid in such as to delay the turnover of the tracer cortisol in plasma. b) The above "dilution" effect of endogenous glucocorticoid surge was very distinct in both the lipophilic fraction and the cortisol fraction of ODS rats, and was less distinct (lipophilic fraction) or insignificant (cortisol fraction) in Wistar rats, a finding to indicate that the observed effect of vitamin C was of physiological significance. c) The responses of the adrenal glands, testes and liver to vitamin C pretreatment were generally more distinct in ODS rats than in Wistar rats, but varied from one organ to another. The significance of the functional linkage between vitamin C and adrenal glucocorticoid, which has been confirmed both in both the humans and rats in our laboratory, was discussed in the light of the historical development of vitamino endocrinology. PMID- 8726813 TI - A review of the antineoplastic action of certain hydrazines and hydrazine containing natural products. AB - The antineoplastic activities of hydrazines and hydrazine-containing natural products are summarized and reviewed. Many of these, including the base compound hydrazine, some of its 79 analogues and 2 hydrazine-containing cultivated mushrooms, exhibited antineoplastic actions in animals and some of them in humans. In addition, a substantial number of hydrazine analogues were further derivatized (altogether 331 derivatives were prepared) and studied to a limited extent for cancer chemotherapeutic activity in animals. Subsequently, only one hydrazine, N-isopropyl-alpha-(2-methylhydrazino)-p-toluamide HCl (procarbazine, natulan) was used extensively in human medicine. Because this drug is a highly carcinogenic substance in animals and since a substantial amount of evidence points to the possibility that it is co-carcinogenic or carcinogenic itself in humans, its use has largely been abandoned. Because well over 80% of the studied hydrazine class of compounds are carcinogenic, therefore, it may not be rewarding to search among them for cancer cure. PMID- 8726815 TI - Effect of ovariectomy on histological change and protein expression in female mouse reproductive tracts. AB - Proliferation of the vagina and uterus in rodents is stimulated by ovarian estrogen, and ovariectomy induces regression of these tracts. In mouse, reproductive tracts, organ weights, vaginal epithelial thickness, the number of epithelial cell layers, and uterine epithelial cell height were significantly reduced 1 to 20 days after ovariectomy when compared with those of intact, mice taken at the estrous stage. No significant differences in those parameters were found between ovariectomized mice implanted with estradiol and intact estrous mice. Vaginal epithelium at estrus was composed of 10-15 layers of epithelial cells with superficial keratinized layers, however, the vaginal epithelium began to regress one day after ovariectomy. Infiltration of leukocytes was encountered in the vaginal epithelium 2-3 days after ovariectomy. Protein expression in the regressing vagina and uterus after ovariectomy was examined by two-dimensional polyacrylamide gel electrophoresis. After ovariectomy, new proteins appeared in the regressing vagina, but not in the regressing uterus. Expressions of 7 and 6 proteins in the vagina and uterus, respectively, reached peak levels 2-3 days after ovariectomy. The expression of ovariectomy-specific proteins and infiltration of leukocytes in vaginal epithelium was inhibited by the injection of cycloheximide, an inhibitor of protein synthesis. These results suggest that some proteins participate in the regression of mouse reproductive tracts after estrogen withdrawal induced by ovariectomy. PMID- 8726816 TI - Anticlastogenicity of vitamin C in vivo against the cytogenetic toxicity of muriate of potash in murine bone marrow cells. AB - The potential in vivo anticlastogenicity of vitamin C was evaluated in murine bone marrow cells against the cytogenetic toxicity of muriate of potash, a synthetic fertilizer. The animals were treated orally once daily for 7 days with muriate of potash (5g/kg) and/or vitamin C (10mg/kg). Metaphase chromosome analyses revealed that muriate of potash induced chromosomal abnormalities were significantly decreased, though still remained higher to control, when vitamin C was administered concurrently. The possible role of vitamin C in mitigating the muriate of potash induced cytogenetic toxicity is discussed. PMID- 8726817 TI - Effect of different vitamin A status on carcinogen-induced DNA damage and repair enzymes in rats. AB - The effect of different vitamin A status on events following DNA damage by hepatocarcinogens was investigated in rats. Formation of single-strand breaks in nuclear DNA induced by aflatoxin B1 and N-nitrosodimethylamine was observed to be more pronounced after vitamin A-deficiency. This enhanced damage was reversed upon vitamin A supplementation. Subsequent to DNA damage, the induction of repair enzymes poly(ADP-ribose) polymerase, DNA polymerase beta and DNA ligase was found to be significantly higher in vitamin A-deficient rats. Vitamin A supplementation brought down the induction to the levels found in rats maintained on normal diet. Vitamin A thus may control carcinogenesis by manipulating molecular events at the initiation stage. PMID- 8726818 TI - Differentiation and IFN gamma regulate WAF1/CIP1 transcription in p53-independent and p53-dependent pathways in epithelial cells. AB - In normal epithelia differentiation and proliferation are regulated by factors involving p53 and related pathways. WAF1/CIP1 protein is believed to inhibit cell growth as a downstream mediator of p53 function. Also, WAF1/CIP1 is identified as a candidate gene linking differentiation signals to G1 arrest. Different epithelia with different keratinizing profiles respond differently to differentiation/proliferation signals, such as calcium or interferon gamma (IFNgamma). Other factors, such as p53 mutation or presence of human papillomaviruses (HPVs) also affect responses to those signals. Since WAF1/CIP1 can be regulated via a p53-dependent or an independent manner, our study aimed to determine: a.) differentiation-related expression of WAF1/CIP1 in keratinocytes, b.) involvement of WAF1/CIP1 in IFNgamma action, and c.) p53-dependence of WAF1/CIP1 transcription under these conditions. The results demonstrated that differentiation increases WAF1/CIP1 transcription in keratinizing but not in nonkeratinizing keratinocytes in a p53-independent manner. IFNgamma upregulated WAF1/CIP1 in keratinizing keratinocytes in a differentiation and p53 dependent manner. Inactivation of wild type p53 by mutation or by presence of HPVs uncouples IFNgamma-mediated WAF1/CIP1 transcription from p53 and from differentiation. Our data suggest a mechanism which operates via WAF1/CIP1 in keratinocytes and regulates epithelial differentiation/proliferation under different physiological conditions. PMID- 8726819 TI - Esophageal Candida infection and adherence mechanisms in the nonimmunocompromised rabbit. AB - Candida infection of the esophagus has been reported not only in immunocompromised hosts but also in healthy individuals. However, its mechanisms of action in healthy individuals have not been clarified. Our previous study suggested that physical contact was an important factor for the adherence of Candida albicans. The aim of the present study was to test our hypothesis and clarify the adherence mechanisms. Suspensions of Candida albicans cells were given to rabbits in drinking water without the use of immunosuppressive drugs and/or antibiotics, and the esophagus was examined. Candidial lesions were observed in 14 of 15 rabbits given the suspensions held in water with and without 30% sucrose for 13 days. The number of Candida albicans cells adhering to the esophagus per square millimeter by subepithelial cell insertion was significantly larger than that adhering by attachment. These results indicate that adherence of Candida albicans to the esophagus occurs by sustained physical contact alone under a nonimmunosuppressive state, and that subepithelial cell insertion results in greater attachment on adherence. Our findings provide a clue that may help clarify the mechanism of Candida infection in healthy individuals. PMID- 8726820 TI - Role of endogenous substance P in ethanol-induced mucosal damage in the rat stomach. AB - To determine the role of endogenous substance P in ethanol-induced mucosal damage, two experiments were performed. In the first experiment, the stomachs of anesthetized rats were doubly cannulated and gastric damage was induced with 5ml of 30% ethanol in the gastric lumen. The damage was ameliorated by pretreatment with capsaicin (0.16 and 1.6 mM) and spantide (100 mg/kg, i.v.). In the second experiment, the gastric mucosa of these rats was perfused with physiological saline containing pepstatin (10 microliters/ml). Endogenous substance P (SP) in the perfusate was measured by enzyme immunoassay (EIA). The peak SP levels were increased by capsaicin (0.16-1.6 mM) in a concentration-dependent manner. Perfusion with 50% ethanol for 5 min increased the SP levels approximately threefold. Perfusion with 1.6 mM capsaicin, followed by 50% ethanol, reduced the injured area to about one-quarter of the original injured area. The peak SP levels during perfusion with 50% ethanol after pretreatment with 1.6 mM capsaicin did not differ from those observed after vehicle pretreatment (control). The area under the curve for SP release during 50% ethanol perfusion after vehicle perfusion was not reduced by previous perfusion with 1.6 mM capsaicin followed by 50% ethanol, indicating that the prevention of ethanol-induced injury by capsaicin may be due to excess amounts of different neuropeptides released simultaneously. PMID- 8726821 TI - Single oral dose of cisapride accelerates gastric antral emptying in healthy humans: an ultrasonographic study. AB - An ultrasonographic study of ten healthy volunteers was carried out to evaluate the effect of cisapride on gastric antral emptying. More than 1 week after the measurement of the baseline emptying rate, cisapride was given at a single oral dose of 5 mg 30 minutes before intake of a balanced liquid test meal (5 ml/kg body weight). To determine the time to half emptying (T1/2), an exponential curve was extrapolated for the elimination phase of the gastroantral sagittal cross sectional area plotted against time. The T1/2 was reduced by 18.5% after cisapride, from 62.6 +/- 4.3 to 51.0 +/- 4.4 min (P = 0.0284). We conclude that a single oral dose of 5 mg of cisapride significantly accelerates the gastric antral emptying rate in healthy humans. PMID- 8726822 TI - Detection of Helicobacter pylori by polymerase chain reaction assay using gastric biopsy specimens taken for CLOtest. AB - The concordance rate between a rapid urease test (CLOtest) and polymerase chain reaction (PCR) assay for the detection of Helicobacter pylori in gastric biopsy samples was investigated. To avoid the bias produced by patchy distribution of the organism in the stomach, the samples used for these two tests were not obtained from two different sites of the antrum. Instead, the PCR assay was performed with the the same biopsy sample that was taken for the CLOtest. Among 82 biopsy samples included for this study, 56 were positive and 26 were negative by CLOtest. Of the 56 CLOtest-positive samples, 52 (93%) were also positive by PCR assay, and of the 26 CLOtest-negative samples, 20 (78%) were negative by PCR assay. The total concordance rate of these two tests was 87.6%. Of the 4 cases with CLOtest-positive and PCR-negative results, 3 had been treated with long-term H2 blockers. Of the 6 patients with CLOtest-negative and PCR-positive results, 4 suffered from recurrent or poorly healing duodenal ulcers. Interestingly, a significantly lower density of the PCR products was observed during electrophoresis analysis for all the 6 cases, presumably due to a small number of H. pylori in these samples. These results indicated that PCR might be used as a complementary assay for CLOtest. False negative results by CLOtest might occur when only a small amount of H. pylori was present in the samples, which could be detected by subsequent PCR assays using the same biopsy specimens. The clinical significance of such CLOtest-negative and PCR-positive cases requires further study. PMID- 8726823 TI - Gastroduodenal manifestations in patients with skeletal fluorosis. AB - A prospective case-controlled study was performed to evaluate the gastrointestinal symptoms and mucosal abnormalities occurring in patients with osteofluorosis. Ten patients with documented osteofluorosis and ten age- and sex matched healthy volunteers were included in the study. Clinical evaluation, real time ultrasound, and upper gastrointestinal endoscopy and biopsy from the gastric antrum and duodenum were performed in all subjects. The biopsies were subjected to a rapid urease test and light and electron microscopic examinations. Ionic fluoride levels were estimated in the drinking water, serum, and urine using an ION 85 ion analyzer. All patients with osteofluorosis had gastrointestinal symptoms, the most common being abdominal pain. Endoscopic abnormalities were found in seven patients with osteofluorosis. In all 7 of these patients, chronic atrophic gastritis was seen on histology. Electron microscopic abnormalities were observed in all 10 patients with osteofluorosis. These included loss of microvilli, cracked-clay appearance, and the presence of surface abrasions on the mucosal cells. None of the control subjects had any clinical symptoms or mucosal abnormalities. It was concluded that gastrointestinal symptoms as well as mucosal abnormalities are common in patients with osteofluorosis. PMID- 8726824 TI - Altered polyamine biosynthesis with aging after massive proximal small bowel resection in rat. AB - We examined the effect of aging on polyamine biosynthesis in the small intestine. Two groups of male Wistar rats (young; 10-week-old, n = 40; old; 24-month-old, n = 40) underwent either a jejunal transection and reanastomosis or 90% proximal small bowel resection. The rats were sacrificed on the 1st, 2nd, 4th, and 7th postoperative day (POD). The mucosa was submitted for histological examination, weighed, and assayed for protein, DNA, RNA, and polyamine content. Ornithine decarboxylase (ODC) activity was measured and ODC mRNA in the mucosa was determined by Northern blot analysis. Compared with the values for wet weight and protein content in old rats, young rats showed significantly higher values for wet weight on the 1st and 2nd POD, and for protein content on the 1st POD, but there were no differences between young and old rats after the 4th POD. The values for ODC activity and ODC mRNA were significantly lower in old rats than in young rats on the 1st POD, but there were no differences between young and old rats after the 2nd POD. The value for putrescine in old rats was significantly lower on the 2nd POD, but was significantly higher on the 4th POD than that in young rats. The present study showed that, in old rats, the residual intestine after small bowel resection preserved sufficient adaptive capacity, but that the adaptive response was decreased. The findings in this study also suggest that a decrease in ODC mRNA expression is involved in the decreased adaptive response that occurs with aging. PMID- 8726825 TI - Pericholangitis in a rabbit colitis model induced by injection of muramyl dipeptide emulsified with a long-chain fatty acid. AB - Rabbit colitis has been induced by injection of muramyl dipeptide emulsified with a long-chain fatty acid. The muramyl dipeptide emulsion was injected submucosally at six portions of the rectum and colon, 10 cm proximal to the anus, using a flexible endoscope. Six rabbits were injected six times every 2 weeks and subsequently killed 2 weeks after the last injection. The histological changes of the colon that occurred in all 6 rabbits were mononuclear cell and histiocyte infiltration with sporadic eosinophils, transmural infiltration, and well maintained goblet cell populations. These changes were different in degree. In 4 of 6 rabbits histological examination of the liver showed pericholangitis and periductal fibrosis mimicking the pericholangitis frequently seen in patients with inflammatory bowel disease. Fibrosis bridging between the portal and portal veins occurred in 2 rabbits, and noncaseating granuloma was seen in 1 rabbit. These histological changes in our model have led to the suggestion that continuous stimulation with bacterial cell wall fragments may be involved in chronic intestinal inflammation and extraintestinal manifestations such as pericholangitis. PMID- 8726826 TI - Growth factor mRNA expression in normal colorectal mucosa and in uninvolved mucosa from ulcerative colitis patients. AB - This study was carried out to investigate the expression of various growth factors (GFs) involved in mucosal healing and thereby to clarify whether there are potential differences in the expression of GFs between normal colonic mucosa and the uninvolved mucosa of ulcerative colitis (UC). GF mRNA was investigated by reverse transcription polymerase chain reaction in colorectal biopsies from 20 normal controls and 15 UC patients. The positive rates (%) for mRNA expression for normal/UC were: epidermal growth factor (EGF) 65/53, transforming growth factor (TGF)-alpha 100/87, TGF-beta 1 60/33, insulin like growth factor-I 45/33, platelet-derived growth factor-A 55/67, basic fibroblast growth factor 0/0, hepatocyte growth factor (HGF) 50/53, EGF receptor 20/27, erb-B2 75/73, and HGF receptor (c-MET) 55/67. Semiquantitation of mRNA showed significantly lower expression of TGF-beta 1 (P < 0.05) in UC. Differences in expression and mRNA levels were not statistically significant for any other GFs. Our results indicate that mucosa in chronic persistent UC has a low basal expression of TGF-beta 1 mRNA, and, since TGF-beta 1 is a multifunctional GF that plays important roles in regulating repair and regeneration following tissue injury, this low expression may be partially responsible for the intractability of the disease. PMID- 8726827 TI - Determining depth of invasion of advanced colorectal cancer using MRI short inversion time inversion recovery sequences. AB - To examine the usefulness of magnetic resonance imaging (MRI) in the preoperative determination of cancerous invasion, we examined 39 patients with advanced colorectal cancer with 0.5T MRI. We employed short inversion time inversion recovery (STIR) sequences, in addition to ordinary spin echo sequences for T1- and T2-weighted images. Preoperatively, the estimated depth of tumor invasion was classified into three grades according to MRI findings, and confirmed on the basis of surgical and histopathologic results. The depth of tumor invasion estimated preoperatively using STIR sequences corresponded well with the surgical and histopathologic results in 85% of the cases. In contrast, assessments based on T1-weighted images corresponded well in only 62% of the cases and T2-weighted images corresponded well in only 64%. PMID- 8726828 TI - Effects of calcium antagonists on hepatic and systemic hemodynamics in awake portal hypertensive rats. AB - The effects of the calcium antagonists diltiazem and nicardipine on portal pressure and splanchnic blood flow were studied in awake, unrestrained portal hypertensive rats. Portal hypertension was induced in rats by partial portal vein ligation. Hemodynamic measurements were done using the radiolabeled microsphere technique. In portal vein-ligated and sham-operated rats, intraarterial diltiazem and nicardipine reduced mean arterial pressure. No significant changes, however, were observed in portal pressure and cardiac index. In portal vein-ligated rats, diltiazem and nicardipine increased portal tributary blood flow. Portal tributary vascular resistance was also significantly decreased. The decrease in the hepatocollateral vascular resistance prevented an increase in portal pressure. In sham-operated rats, these changes were not observed. It is possible that the vascular responses to calcium antagonists are altered in portal vein-ligated rats. These findings demonstrate that the hemodynamic effects of calcium antagonists occur at two levels. First, the increase in portal tributary blood flow appears to be a selective effect on portal tributary vascular resistance. Secondly, the portal pressure does not increase in parallel with the increase in portal tributary blood flow because of a similar reduction in portocollateral vascular resistance. PMID- 8726829 TI - Comparison of the proliferative capacity of adenomatous hyperplasia and well differentiated hepatocellular carcinoma. AB - The identification of nodular lesions, which used to be difficult to differentiate from well differentiated hepatocellular carcinomas, has become frequent since the advent of ultrasonic-guided liver tumor biopsy techniques. This is because adenomatous hyperplasias, which are regarded as putative precancerous lesions, which needed to be differentiated from malignant lesions, often an a nodular. The histological specimens of 52 tumors, obtained from patients with chronic liver injury, were subjected to immunohistological staining for proliferative cell nuclear antigen (PCNA). Findings obtained by this immunohistological technique were compared with those obtained by histological image analysis. Adenomatous hyperplasias with fatty changes or with a high nuclear cytoplasmic ratio on image analysis were frequently PCNA-positive (P < 0.01). These nodular lesions, whose morphological characteristics were similar to those of well differentiated hepatocellular carcinomas, showed a high potential for growth. It seems that PCNA may be a useful indicator for increasing our understanding of the progress from adenomatous hyperplasia to hepatocellular carcinoma. PMID- 8726830 TI - Pathophysiological effect of hepatic ischemia and reperfusion after hepatectomy in dogs with obstructive jaundice, focusing on the effect of coenzyme Q10 and styrene-co-maleic acid superoxide dismutase. AB - The purpose of the present study was to elucidate the effect of hepatic reflow following ischemia on the remnant liver after hepatectomy with occluded hepatic blood inflow in dogs with obstructive jaundice. When 40% hepatectomy was performed with 10-min occlusion of hepatic blood inflow in dogs with obstructive jaundice, the lipid peroxide content in the remnant liver increased significantly, together with a reduction in superoxide dismutase (SOD)-like activity. The levels of endotoxin and beta-N-acetyl hexosaminase (NAH) in peripheral blood also increased. The phagocytic index increased transiently after 30 min, followed by a marked decrease after 3h. Histologically, degeneration and necrosis of the hepatic parenchymal cells were demonstrated, and survival rate at 7 days was only 23.1%. With the administration of coenzyme Q10 (CoQ10) or styrene co-maleic acid SOD (SM-SOD), these phenomena were significantly inhibited, and the survival rate improved. After hepatectomy, Kupffer cells in the remnant liver were activated by increased endotoxin levels in the portal vein, inducing the production of free radicals, which, in turn, damaged the Kupffer cells by reducing endotoxin clearance. Finally, the impaired functional reserve in the remnant liver provoked liver failure. The administration of CoQ10 or SM-SOD prevented the occurrence of these phenomena triggered by the free radicals generated by Kupffer cells, stimulated by endotoxin in the portal vein. PMID- 8726831 TI - Evaluation of patient outcome following sclerotherapy for esophageal varices. AB - After excluding terminally all patients, we evaluated a total of 718 patients treated with endoscopic injection sclerotherapy. They involved 350 episodes of acute hemorrhage and 368 prophylactic procedures in patients with risky varices. The 1-year cumulative survival rate was significantly lower in the acute hemorrhage group than in the prophylactic group (P < 0.05). The difference in survival between the two groups was primarily due to the number of deaths in the first 2 months after sclerotherapy (20.1% vs 0.8%, P < 0.0005). Improvements in the sclerotherapy technique significantly reduced the number of deaths from bleeding (9.3% vs 3.4%, P < 0.05), but not those from liver failure following variceal hemorrhage. Prophylactic EIS is advantageous in the treatment of esophageal varices, i.e. it may prevent deaths from liver failure attributed to variceal hemorrhages. The present study shows that preliminary prevention of variceal hemorrhage provides favorable hemostatic efficacy in patients with risky varices. PMID- 8726832 TI - Therapeutic effect of secretin in patients with jaundice; double-blind placebo controlled multicentric trial. AB - Secretin, a gastrointestinal hormone, has been shown to have a potent choleretic effect. Having already obtained some beneficial effects with secretin in patients with intrahepatic cholestasis, we sought to confirm its effects in a double-blind placebo-controlled study in patients with mild jaundice after acute or during chronic hepatitis, where total bilirubin level was in excess of 4.0 mg/dl for 3 days or more. Patients with primary biliary cirrhosis (PBC), primary sclerosing cholangitis (PSC), and familiar hyperbilirubinemia were excluded from the study. Ninety-three patients were included in this analysis, but the final evaluation covered 69 of them. No statistically significant differences were found in the reduction of serum bilirubin levels between secretin and placebo groups. As a number of patients with liver cirrhosis had been included, the subjects were subdivided into one group with cholestasis in hepatitis and one with liver cirrhosis. In the subgroup of cirrhotic patients who received secretin, serum levels of AST were significantly increased compared with the placebo group. However, since the choleretic effect of secretin is unique, further studies seem to be warranted. PMID- 8726833 TI - Role of endogenous and exogenous cholecystokinin in experimental acute pancreatitis induced in rats by the duodenal loop technique. AB - The role of endogenous cholecystokinin (CCK) release and exogenous CCK-8 administration in the development and progression of acute pancreatitis and in the early recovery phase of acute pancreatitis were investigated in rats with closed duodenal loop (CDL)-induced pancreatitis. The subcutaneous injection of CCK-8 (2 micrograms/kg) stimulated a physiological level of pancreatic enzyme secretion in normal control rats, but did not lead to any biochemical or histological evidence of acute pancreatitis. A higher dose of CCK-8 (8 micrograms/kg), however, did produce both biochemical and histological evidence of acute pancreatitis in the normal control rats. When 2 micrograms/kg of CCK-8 was injected subcutaneously in rats 6 and 12 h after the creation of the CDL, neither the biochemical nor the histological findings of acute pancreatitis showed any progression compared with the changes in controls given no CCK-8. Serum CCK levels, measured by radio-immunoassay, increased significantly from mean levels of 5.39 pg/ml (+/- 0.95 SD) before creation of the CDL to 42.06 pg/ml (+/- 2.27 SD) 6 h after, and 41.95 pg/ml (+/- 1.88 SD) 12 h after its creation (P < 0.01). The difference between serum CCK levels at 6 and 12 h was not statistically significant. Following the release of the loop, serum CCK levels decreased gradually, especially in rats in which the loop was released 6 h after being created. Although no marked biochemical and histological changes of acute pancreatitis were observed following the administration of 2 micrograms/kg of CCK 8 to rats upon release of the loop 6 h and 12 h after its creation, a higher dose of CCK-8 (8 micrograms/kg) in these rats adversely affected both the biochemical and histological findings of acute pancreatitis. Based on these findings, it was concluded that neither endogenous CCK release, as a result of the CDL, nor physiological stimulation of the pancreas by exogenous CCK-8 administration, caused progression from edematous to hemorrhagic acute pancreatitis, and neither CCK treatment had any adverse effect on the early recovery phase of CDL-induced acute pancreatitis. A pharmacological dose of CCK, however, exacerbated the acute pancreatitis, even in the early recovery stage. PMID- 8726834 TI - Role of endogenous platelet-activating factor in the regulation of pancreatic blood flow during caerulein stimulation. AB - The role of endogenous platelet-activating factor (PAF) in the control of pancreatic blood flow during caerulein stimulation was investigated. Pancreatic blood flow in anesthetized rats was measured continuously by laser Doppler flowmetry for 2 h during the intravenous infusion of caerulein (0.25 micrograms/kg per h). Pancreatic blood flow showed a gradual, consistent, and significant increase, reaching 114.2 +/- 2.3% of the basal value after 120 min. Changes in pancreatic blood flow induced by caerulein were completely inhibited by a cholecystokinin (CCK) antagonist (loxiglumide, 5 mg/ kg per h, i.v.) and by a specific PAF antagonist (CV-6209, 1 mg/kg, i.v.-bolus). Systemic blood pressure remained stable in all groups. These results suggest an important role of endogenously yielded PAF in regulating pancreatic blood flow during caerulein stimulation to the pancreas. PMID- 8726835 TI - Gastric duplication cyst: evaluation by endoscopic ultrasonography and magnetic resonance imaging. AB - Endoscopic ultrasonography (EUS) and magnetic resonance imaging (MRI) are becoming popular methods for examining tumorous lesions along the upper gastrointestinal tract. Though duplication cysts are uncommon, EUS findings from gastric duplication cysts have accumulated and proven very useful for preoperative diagnosis. There have been few reports, however, concerning MRI findings from these cysts. We report herein the case of a 25-year-old man with a gastric duplication cyst. EUS demonstrated a cystic mass adjacent to the fourth layer of the stomach wall. MRI revealed a cyst containing low signal-intensity fluid and high signal-intensity fluid separated by levels. In addition to the characteristic findings from preoperative examinations, the unique histological findings from the cyst are also described. PMID- 8726836 TI - Laparoscopically assisted ileocecal resection for Crohn's disease associated with intestinal stenosis and ileovesical fistula. AB - We describe a 22-year-old man with Crohn's ileocolitis accompanied by intestinal stenosis and ileovesical fistula in whom laparoscopically-assisted surgery was successfully performed after thorough nutritional therapy. Laparoscopic procedures are characterized by minimal access and minimal invasion, features which can contribute to the early recovery of patients who undergo surgery. It is suggested that laparoscopic (or laparoscopically-assisted) surgery after strict nutritional therapy can be effective in the treatment of patients with Crohn's disease who have intestinal complications. PMID- 8726837 TI - Coexistence of choriocarcinoma and adenocarcinoma in the rectum: molecular aspects. AB - Choriocarcinoma, a malignant tumor of usually placental origin, in divided into two groups; the gestational and non-gestational types, the latter being rare. Non gestational choriocarcinoma occurs in the lung, mediastinum, kidney, stomach, and small intestine, but rarely appears in the large intestine. We treated a 29-year old woman with choriocarcinoma of the rectum with adenocarcinoma. Despite the rarity of the condition and the obscurity of the histogenesis, reports of similar cases and the occurrence of the tumors in the digestive tract suggest that the condition constitutes a clinical entity of a digestive tumor. PMID- 8726838 TI - Primary malignant melanoma of the rectum. AB - We present a case of primary malignant melanoma arising in the rectum of a 71 year-old woman who had presented with intermittent rectal bleeding following bowel movement 2 months previously. Digital and sigmoidoscopic examination of the rectum revealed a 3 x 2 x 1 cm exophytic tumor with a granular surface in the rectum 35 mm from the dentate line. No increased pigmentation was detected. Biopsy revealed degenerated mucosa accompanied by severe infiltration of inflammatory cells, but no malignant cells. The tumor and normal mucosa surrounding the tumor was excised 12 days after the biopsy. Light microscopy revealed the tumor to consist of malignant melanocytes and showed that the tumor was surrounded by normal mucosa. Immunohistochemical staining with antimelanoma antibody and HMB-45 substuntiated the diagnosis. Clinical and laboratory examination excluded the presence of melanoma at sites other than rectum. The patient refused an abdomino-perineal resection of the rectum and combination chemotherapy was performed. She died 18 months after the initial operation due to local recurrences and metastases to pelvic lymph nodes, liver, and lung. PMID- 8726839 TI - Liver cell adenoma in a young man with elevated serum PIVKA-II level. AB - A 21-year-old man was referred to our hospital because of a liver mass lesion detected by abdominal ultrasonography. He had received no hormonal treatment. Physical examinations revealed no abnormalities, and laboratory data, including hepatic function test results, were within normal ranges, with the exception of elevated levels of serum protein induced by vitamin K absence or antagonist (PIVKA)-II (2.2 AU/ml). Abdominal ultrasonography revealed a hyperechoic mass lesion measuring 10 x 10 cm, with hypoechoic areas located in the right posterior segment of the liver. A low-density area and a hypervascular area were detected in the right posterior segment of the liver by computed tomography and celiac angiography, respectively. As hepatocellular carcinoma could not be completely excluded, the tumor was resected. The tissue consisted of sheets of tumor cells with eosinophilic cytoplasm and round nuclei showing a thin trabecular pattern, and these histological findings indicated liver cell adenoma. After resection of the tumor, serum PIVKA-II returned to the normal level. PMID- 8726840 TI - Ciliated foregut cyst in cirrhotic liver. AB - We encountered a patient with a ciliated hepatic foregut cyst with accompanying liver cirrhosis, which was hard to distinguish from well-differentiated hepatocellular carcinoma. A lesion 2 cm in diameter was found in the subcapsular region of the medial segment of the liver. It was slightly hypoechoic on ultrasonography, of high attenuation on nonenhanced computed tomography (CT), of high intensity on T1-weighted spin echo images of magnetic resonance imaging (MRI), and of isointensity on T2-weighted spin echo images. It was not enhanced in the arterial phase images of MRI, and was shown as a complete perfusion defect on CT arterial portography. The cyst was enucleated and found to be filled with bloody mucinous fluid. PMID- 8726841 TI - Hepatocellular carcinoma associated with anabolic steroid therapy: report of a case and review of the Japanese literature. AB - We report herein the case of a 35-year-old woman with aplastic anemia who developed hepatocellular carcinoma after long-term therapy with oxymetholone. She was treated with 60 mg/day of oxymetholone for 3 years (total dose 64.8 g). Alpha fetoprotein, hepatitis B surface antigen, and hepatitis C antibody were all negative, but serum titers of carcinoembryonic antigen and carbohydrate antigen were elevated. Lateral segmentectomy of the liver was performed. The histopathological findings were compatible with those of multiple hepatocellular carcinoma without liver cirrhosis. Three years since the operation, the patient is doing well and no signs of tumor recurrence have been detected. According to our review of Japanese cases of hepatocellular carcinoma associated with anabolic steroid therapy, in all instances the tumors developed after long-term administration of anabolic steroids for hematologic diseases. In patients under long-term anabolic steroid therapy, routine screening of the liver by ultrasonography and computed tomography should be performed to detect liver tumors in the early stages. PMID- 8726843 TI - Ampullary somatostatinoma in a patient with von Recklinghausen's disease. AB - We report a case of somatostatinoma of the ampulla of Vater associated with von Recklinghausen's disease in a 44-year-old woman. On admission the patient was jaundiced, and percutaneous Cholangio-drainage was performed. Cholangiography revealed stenosis of the common bile duct at the lower end Duodenoscopy showed a yellowish tumor of the ampulla of Vater, and the biopsy specimens showed no malignant cells. Pylorus-preserving pancreaticoduo-denectomy was performed. Histologically, the tumor was composed of small round cells with a solid or trabecular pattern and with multiple psammoma bodies. Immunohistochemical examination showed that the tumor cells stained for somatostatin. Genomic examination showed neither K-ras nor p53 gene mutations of the resected specimen. PMID- 8726842 TI - Mucin-producing adenoma associated with pancreas divisum and hepatic hilar carcinoma: an autopsy case. AB - We present the autopsy case of an 82-year-old Japanese woman with a mucin producing adenoma accompanied by pancreas divisum and a hepatic hilar carcinoma. She had suffered from a cholangiocellular carcinoma at the hepatic hilus for 2 months, which was treated with radiation and chemotherapy. She did not complain of any abdominal pain. Obstructive jaundice deteriorated despite percutaneous transhepatic bile duct drainage, and she died of hepatic insufficiency. At autopsy, a hepatic tumor was confirmed to have caused severe obstructive jaundice. Histological examinations showed moderately to poorly differentiated cholangiocellular adenocarcinoma with squamous metaplasia, probably due to radiation. A yellowish mucinous tumor was found in the head of the pancreas near the minor papilla. It consisted of multiple rice-sized cystic lesions with thin septa. Although it had no capsule, its margin was clear. Neither a wide opening of the major or minor papilla nor mucous drainage was observed. Gross examinations revealed unfused pancreatic ducts. The slightly dilatated dorsal duct and a branch of the mildly dilatated ventral duct showed tumor involvement. Histological examinations showed mild atypia of the epithelia, and this pancreatic tumor was diagnosed as branch duct-type mucin-producing adenoma with postradiation dysplasia. The combination of a mucin-producing tumor and pancreas divisum is rare, and this is only the third reported case. PMID- 8726844 TI - Floating gallbladder associated with histologically distinct double cancers. AB - A case of unusually hypermobile floating gallbladder in a 79-year-old woman with histologically distinct double cancers of the gallbladder is described. The patient presented with an abdominal cystic mass, which was palpable with easy mobility from the right lower quadrant practically to the left upper quadrant. Exploratory laparotomy was performed and the cystic mass was found to be a floating gallbladder. The cystic duct was elongated and obstructed, and had a long mesentery. After the operation, latent double cancers of the gallbladder were discovered on histopathological examination. The obstruction of the cystic duct was due to chronic inflammation and had resulted in hydrops of the gallbladder. This was suspected to have played an important role in the carcinogenesis. We believe that this is the first report of a floating gallbladder associated with double gallbladder cancers. PMID- 8726845 TI - Gallbladder cancer associated with cholesterosis. AB - We report herein two cases of carcinoma in situ of the gallbladder associated with cholesterosis. The patient in case 1 was an 81-year-old man who underwent a cholecystectomy for cholelithiasis. The resected specimens revealed gallbladder cancer in the fundus which was diagnosed histologically as mucinous carcinoma. Other findings included 13-mm, 12-mm, and 5-mm polypoid lesions in the neck of the gallbladder which macroscopically appeared to be cholesterol polyps, but histologically demonstrated carcinoma in situ with cholesterosis. The patient in case 2 was a 76-year-old man in whom ultrasonography revealed a highly echogenic, elevated lesion in the gallbladder. Cholecystectomy was performed, and a 33 x 28 mm papillary, elevated lesion with cholesterosis was resected from the neck of the gallbladder. Histologically, this was demonstrated to be papillary adenocarcinoma in situ with cholesterosis surrounded by glandular dysplasia. The distribution of the carcinomas and cholesterosis in both of these patients suggests that the adenoma or carcinoma of the gallbladder had occurred first. Then, the tumor epithelium absorbed cholesterol from the bile, and foamy cells were produced. Thus, when treating cholesterol polyps, it should be remembered that it is often difficult to distinguish between cholesterol polyp and gallbladder cancer with cholesterosis. PMID- 8726847 TI - Expression of bone marrow stromal cell specific antigen during murine development: its expression in embryonic hematopoietic tissues as well as in other developing tissues. AB - Monoclonal antibody R4-A9 demonstrated specificity for a cell surface antigen of stromal cells in murine bone marrow and spleen. In order to identify patterns of expression that may elucidate the potential role of R4-A9 antigen, the developmental expression of this antigen in mouse embryos from 8 days post-coitum to 5 days post-partum was investigated by immunohistochemistry. At an early developmental stage, weak staining for R4-A9 antigen could be detected in the yolk sac. At later stages, strong staining of this antigen was detected predominantly in the embryonic liver, the main site of embryonic hematopoiesis. However, concomitant with the decreased staining in the liver, increased expression of this antigen was observed in bone marrow and spleen. Therefore, the changes in expression in those hematopoietic tissues suggest that its expression is coordinately regulated during the developmental stage of the sites of embryonic hematopoiesis. Compared with the distribution of R4-A9 antigen in adult tissues as previously reported, the expression of this antigen in fetal tissues was more widespread during the period of organogenesis, and was most abundant in other developing tissues, including the heart, skin, and lung. In contrast, fetal expression detected in hematopoietic and other developing tissues was lost after birth. These results taken together show a marked gradient of R4-A9 antigen expression, with the highest level at the peak of organ development, raising the possibility that this molecule may act as a growth/differentiation factor both in hematopoietic and other developing tissues in a fetus. PMID- 8726846 TI - Bile secretion--models, mechanisms, and malfunctions. A perspective on the development of modern cellular and molecular concepts of bile secretion and cholestasis. PMID- 8726848 TI - Increased cell proliferation and transforming growth factor-alpha (TGF alpha) in the gall-bladder epithelium of patients with pancreaticobiliary maljunction. AB - Pancreaticobiliary maljunction is associated frequently with gall-bladder carcinoma. Although increased turnover of the gall-bladder epithelium in patients with pancreaticobiliary maljunction is thought to predispose to carcinogenesis, there is little data to confirm this hypothesis. In addition, no previously published study has addressed the process underlying cell proliferation. In this study, cell kinetics were first evaluated using two methods, proliferating cell nuclear antigen (PCNA) immunohistochemical staining and argyrophilic nucleolar organizer region (AgNOR) staining. Second, immunohistochemistry was used to investigate the expression of transforming growth factor-alpha (TGF alpha), a potential regulator of cell proliferation in the gall-bladder. The gall-bladders of 11 patients with pancreaticobiliary maljunction were studied, and 11 gall bladders removed from patients during other surgery were used as controls. The number of PCNA-positive cells and the number of AgNOR per nucleus were significantly greater in the gall-bladders of patients with pancreaticobiliary maljunction than in the control gall-bladders. The expression of TGF alpha was also significantly greater in the gallbladders of patients with pancreaticobiliary maljunction than in the control gall-bladders. In conclusion, these results suggest that the increased TGF alpha expression induced by pancreaticobiliary maljunction promotes proliferation of the gall-bladder epithelium, which may lead to carcinogenesis. PMID- 8726849 TI - Cellular differentiation and development of pyloric mucosal metaplasia in the human gall-bladder. AB - The development of pyloric mucosal metaplasia (PMM) with regard to cellular differentiation in the human gallbladder was studied by mucin staining (paradoxical concanavalin A (Con A), galactose oxidase-Schiff (GOS) and alcian blue (pH 2.5) PAS (AB-PAS) and immunohistochemistry (pepsinogen II (PgII) and SH 9, and proliferating cell nuclear antigen (PCNA). PMM was divided into three stages of development by three-dimensional (3D) computer graphic reconstruction analysis. In the early stage, a transitional zone of PCNA positive cells was observed between areas of SH-9 and/or GOS reactive cells and class III and/or Pg II positive cells in flat monolayered epithelium. In the middle stage, shallow pits became apparent as areas enlarged, with these becoming deeper in the advanced stage, whereby SH-9 and/or GOS reactive cells and class III and/or Pg II positive cells were observed at the upper and lower portion of the pits, respectively, with PCNA-positive cells forming a narrow zone between the two cell populations. Consequently, the structure of PMM gradually resembles that of the normal gastric pyloric mucosa. PMID- 8726850 TI - Regenerative pattern of liver cells in primary biliary cirrhosis, alcoholic cirrhosis, posthepatitic cirrhosis (HBV-related) and hepatocellular carcinoma: comparative analysis by computerized morphometry. AB - Computerized morphometrical measurements were made of liver cells and their nuclei taken from biopsy specimens of primary biliary cirrhosis (PBC), alcoholic cirrhosis, posthepatitic cirrhosis (HBV-related), and hepatocellular carcinoma (HCC). The specimens were stained with hematoxylin-eosin (HE), Mallory's stain for collagen fibers, orcein method, periodic acid Schiff (PAS) reaction, and silver impregnation. Light microscopic views were then selected and original liver cells were magnified x 1000. The size of liver cell nuclei, distance between corresponding liver cell nuclei and distribution pattern of hepatocytes were calculated by computer. Variation in regenerative activity among the four disease groups was noted. Regenerative features of liver cells were mild in degree in PBC. In alcoholic cirrhosis, regenerative features of liver cells were less prominent than in posthepatitic cirrhosis. In posthepatitic cirrhosis, regenerative liver cells were well developed, showing remarkable pleomorphism of liver cell nuclei and expansive arrangement of liver cell cords. This tendency towards regenerative activity suggests that the possibility of HCC occurring is greater in posthepatitic cirrhosis than in PBC or alcoholic cirrhosis. It was concluded that morphologically, there is a greater possibility of occurrence of HCC in posthepatitic cirrhosis than in any other type of cirrhosis, because of its high regenerative hepatocytic activity. Also etiological factors of liver diseases are more important in the development of liver cell regeneration. Furthermore, regenerative activity can be measured by computerized morphometry as an established methodology. PMID- 8726851 TI - Different expression of Bcl-2 protein in gastric adenomas and carcinomas. AB - In order to cast light on the possible role of bcl-2 protein (Bcl-2) expression in gastric tumorigenesis, 33 cases of gastric adenomas and carcinomas originating from the same stomachs were immunohistochemically investigated for Bcl-2 protein (Bcl-2) expression, accumulation of p53 protein and cell proliferation as determined by the Ki-67 labeling index (LI). Bcl-2 expression was detected in 24/33 (72.7%) adenomas and in 6/33 (18.2%) carcinomas, the difference being statistically significant (P = 0.0001). Only 4 of 33 (12.1%) cases exhibited expression in both adenoma and carcinoma lesions in the same stomachs. Immunoreactivity was decreased in areas of cellular and structural atypia in adenoma lesions (P < 0.008), and appeared to be positively linked to the tumor progression and the degree of differentation in carcinomas, although it did not reach statistical significance. Accumulation of p53 protein was rare in the adenomas but was found in 15/33 (45.5%) of carcinoma lesions, with a significant dissociation from Bcl-2 immunoreactivity. No apparent relation between Ki-67 LI and either adenoma grading or carcinoma typing was noted, although average Ki-67 LI of the highest labeling areas in carcinomas was statistically higher than in adenomas (P = 0.0001). These results indicate that the regulation of Bcl-2 expression may differ between gastric adenomas and carcinomas, may be correlated with tumor differentiative features. In addition, p53 accumulation may play an important role in the onset of malignancy. PMID- 8726852 TI - Postmenopausal intra-abdominal desmoplastic small cell tumor. AB - Intra-abdominal desmoplastic small cell tumor (DSCT) usually occurs in infants and young male adults. A case of DSCT occurring in a 60 year old female is described. No other apparent primary origin was detected. A mesocolon tumor, measuring 23 x 12 x 10 cm, was composed predominantly of round to spindle cells which showed epithelioid- and focally sarcomatous arrangements. Immunohistochemically, the tumor cells showed perinuclear dot-like staining of CAM5.2, many cells expressed HHF35, and some cells contained vimentin, epithelial membrane antigen, desmin, alpha-smooth muscle actin, neuron-specific enolase, or Leu 7. Electron microscopic examination showed that the tumor cells had mesenchymal-fibroblastic features. The tumor had an aneuploid DNA content with high S-phase fraction. The patient, who was treated with adjuvant chemotherapy, was alive, having had three recurrences in 36 months. In the second and third recurrent lesions, increased cellular atypia and fascicular arrangements of spindle cells were observed. DSCT should be included in differential diagnoses of post-menopausal pelvic tumors which show light-microscopically and immunohistochemically divergent phenotypes. PMID- 8726853 TI - Oncocytic hepatocellular carcinoma with numerous globular hyaline bodies. AB - Two well circumscribed tumors, oncocytic and non-oncocytic, were removed from the non-cirrhotic liver of a 67 year old male. The large oncocytic tumor (OCT), occupying the entire left lobe, was multilobulated with focal coagulation necrosis and areas of hemorrhage. Light microscopy revealed that it consisted of exclusively large, granular oxyphilic cells with moderate nuclear atypia and occasional mitotic figures, which were trabecular and/or pseudoglandular in structure, but no lamellar fibrosis was seen. Characteristically, the OCT cells included numerous globular hyaline bodies (GHB) of various sizes which were stained red with acid fuchsin and deep blue or magenta with phosphotungstic acid hematoxylin (PTAH), but negative for periodic acid Schiff (PAS), orcein, rhodamine and Grimelius methods. Immunohistochemically, alpha-fetoprotein (AFP), alpha-1-antitrypsin, alpha-1-antichymotrypsin, fibrinogen and ferritin were all negative. On ultrastructural examination, tumor cells were mitochondria-rich, including electron dense, ovoid or polyhedral inclusions, with the delineated membrane identical with that of the GHB. In contrast, the small tumor in the right lobe (Segment 7) was a solid adenoma with no oncocytic transition. Based on these findings, it was postulated that OCT consists of heterogenous proliferation of mitochondria-rich hepatocytes which tend to induce lysosomal GHB closely associated with mitochondrial abnormalities. PMID- 8726854 TI - Renal primitive neuroectodermal tumor: an immunohistochemical and cytogenetic analysis. AB - The cytogenetic and morphologic characteristics of a case with a primitive neuroectodermal tumor (PNET) arising from the left kidney in a 22 year old man are presented. The patient was detected as having a left renal mass with a tumor embolus in the inferior vena cava and multiple pulmonary metastases. A radical nephrectomy with tumor embolectomy of the inferior vena cava, along with a resection of the pulmonary nodules were performed. Histologic examination revealed a dense proliferation of small round cells with many Homer-Wright type rosettes and perivascular pseudorosettes. Immunohistochemically, the tumor cells stained strongly positive for HBA71(p30/32MIC2), a surface glycoprotein specific to PNET and Ewing's sarcoma. In addition, the tumor cells expressed several neural markers (neuron specific enolase, neurofilament, synaptophysin, and Leu-7) and vimentin, while the epithelial, muscular, and lymphocytic markers were negative in the tumor cells. Cytogenetic analysis of cultured tumor cells showed a reciprocal translocation t(11;22)(q24;q12) that is considered to be specific to PNET and Ewing's sarcoma. In conclusion, this case suggested that a karyotyping analysis is a useful diagnostic tool for renal PNET, and it may therefore be utilized to help distinguish between difficult cases of small round cell tumors and Wilms' tumor of the kidney. PMID- 8726855 TI - Granular cell tumor expressing myogenic markers in the prostate. AB - A granular cell tumor in the prostate of a 72 year old male is reported. The tumor nest was composed of large polygonal tumor cells, possessing cytoplasmic eosinophilic granules which were positive with PAS stain and resistant to diastase digestion. Immunohistochemically, most tumor cells were strongly positive with anti-alpha-smooth muscle actin and -neuron specific enolase antibodies. Only a few scattered tumor cells were reacted with anti-S-100 protein antibody. These findings might suggest smooth muscle differentiation of this case, and provide an additional insight into the histogenesis of granular cell tumors, especially in their heterogeneity. PMID- 8726857 TI - Glomerulonephritis with predominant paramesangial IgG deposition. AB - The new clinicopathological entity, immunoglobin G (IgG)-associated mesangial proliferative glomerulonephritis (GN), has been reported recently, but serial renal biopsies were not performed in the cases reported. The findings of three serial renal biopsies in a pediatric case with IgG-associated GN and paramesangial deposits are reported. Microscopic hematuria was found incidentally at the age of 8 years and the hematuria often worsened transiently during periods of upper respiratory infections. The patient was treated mainly with dipyridamole. The third biopsy showed that both paramesangial hemispherical deposits and predominant mesangial IgG deposits had increased, while mesangial cell proliferation had markedly decreased. These serial biopsy findings suggest that IgG-associated GN with microscopic hematuria and slight proteinuria may be characterized by a relatively benign histological and clinical course, as described in recent reports. PMID- 8726856 TI - Granulomatous cholangitis in chronic hepatitis C: a new diagnostic problem in liver pathology. AB - A case of chronic hepatitis C at the pre-cirrhotic stage complicated with hepatocellular carcinoma is reported. The patient, a 64 year old female, showed elevated levels of serum alkaline phosphatase and immunoglobulin M. Antimitochondrial antibodies were negative by indirect immunofluorescence. Western blotting using beef heart mitochondria and recombinant polypeptides coding for mitochondrial antigens revealed that the patient's serum was positive only for the E2-subunit of the branched chain ketoacid dehydrogenase complex. In the non-neoplastic liver, chronic non-suppurative cholangitis surrounded by epithelioid granuloma, resembling the granulomatous destructive cholangitis of primary biliary cirrhosis, was found. The damaged bile ducts were immunohistochemically minimally positive or ambiguous for HLA-DR, and their expression of the E2-subunit of the pyruvate dehydrogenase complex E2 (PDC-E2) was diffuse or granular, and not typical of primary biliary cirrhosis. There was no bile duct loss, and orcein-positive copper binding granules reflecting chronic cholestasis were negative in periportal hepatocytes. The overall features in this case were consistent with primary biliary cirrhosis presenting an infrequent profile of antimitochondrial antibodies and atypical expression of HLA-DR and PDC E2 on biliary epithelial cells, with late superimposition on chronic hepatitis C. However, it is also possible that this is a case of chronic hepatitis C with hepatitis-associated bile duct damage accompanied by granulomatous reaction. Either way, this case raises new diagnostic issues in the differential diagnosis of chronic liver diseases presented with granulomatous cholangitis. PMID- 8726858 TI - Control and management of ovarian follicles in cattle to optimize fertility. AB - Experiments were designed to elucidate the control of ovarian follicle turnover and the impact of follicular dynamics on the subsequent fertility of dairy cattle. An experimental model was established to examine the interrelationships of gene expression for steroid enzymes, the insulin-like growth factor system and inhibin production as associated with follicle selection, dominance and atresia. Follicular dynamics during the postpartum period and the oestrous cycle are shown to be altered markedly by the metabolic demands of lactation. The feeding of ruminally-inert fat stimulated follicular development and improved reproductive performance. The development of persistent follicles during oestrus synchronization causes a reduction in fertility that can be corrected by recruitment and selection of a new ovulatory follicle after the injection of a gonadotrophin-releasing hormone agonist. Present systems of oestrus synchronization need to consider both synchronization of follicular development and corpus luteal regression in order to optimize fertility. With current systems manipulating follicle development, the potential to implement a timed insemination programme to improve reproductive management exists. Ovulation of the first-wave dominant follicle with human chorionic gonadotrophin provides a means to markedly enhance concentrations of plasma progesterone in the luteal phase. PMID- 8726859 TI - Optimization of a continuous real-time computerized semen analysis system for ram sperm motility assessment, and evaluation of four methods of semen preparation. AB - Sampling conditions that affect the biological validity of computer-assisted analysis of ram sperm motion were examined using a continuous real-time computerized semen analysis system (Hobson Sperm Tracker). Search radius (SR, 10 settings) and minimum track point (MTP, 10 settings) were varied factorially to evaluate their effects on the inclusion of sperm subpopulations within derived datasets. Low SR (< 12 microns) or high MTP values (> 26 frames) precluded measurements of rapidly moving cells, whereas high SR (> 17 microns) and low MTP settings (< 22 frames) led to erroneous tracking and poor data quality. Suitable settings for these set-up parameters were derived and tested for their biological consistency with four methods of preparing ram semen for computerized analysis. The preparation techniques tested were: centrifugation through sucrose-based Ficoll and Percoll media, a swim-up technique and simple dilution in Tyrode's media. The "selective' Percoll and swim-up methods generated sperm populations with significantly higher linear velocities and a lower tendency to deviate from linear trajectories than from either the Ficoll method or dilution technique. Deleterious effects of centrifugation were evident, particularly on sperm survival in vitro over several hours. It is concluded that computer-assisted semen analysis provides useful information about the behaviour of ram spermatozoa in vitro, but the measurement conditions must be defined carefully. PMID- 8726860 TI - Significant decrease in parathyroid hormone-related protein concentrations in amniotic fluid with labour at term but not preterm. AB - It has been determined whether amniotic fluid concentrations of parathyroid hormone-related protein (PTHrP) change with labour. An evaluation of which cells from intrauterine tissues might produce PTHrP has also been conducted. Amniotic fluid was obtained by amniocentesis from women: (1) at term, not in labour; (2) in normal term labour; (3) in preterm labour, undelivered within one week; (4) in preterm labour, delivered within one week; (5) in preterm labour associated with clinical chorioamnionitis; and (6) who were gestation-matched controls for chorioamnionitis patients-women in this group were similar to those in Group 4 but were different patients. Amnion, chorion, and decidual cells were grown by standard techniques and incubated with interleukin-1 beta (IL-1 beta). PTHrP was assayed in duplicate samples of amniotic fluid or tissue culture media using an immunoradiometric assay. There was a significant reduction in amniotic fluid concentrations of PTHrP during labour at term. Preterm labour was not associated with significant changes in amniotic fluid concentrations of PTHrP although a trend for reduced concentrations was observed. Amnion and chorion produced measurable quantities of PTHrP and rates of production were increased by treatment with IL-1 beta. Decidual cells did not produce detectable amounts of PTHrP. Hence, labour at term is associated with a decrease in amniotic fluid PTHrP concentrations that may reflect reduced amnion production, which in turn may play a permissive or active role in the mechanism(s) of parturition. These data support the view that the mechanisms that control term and preterm labour may be regulated differently. PMID- 8726861 TI - The relationship between chromosomal abnormality in the human preimplantation embryo and development in vitro. AB - The relationship between the survival of the human preimplantation embryo in vitro and chromosomal abnormality was investigated by cytogenetic analysis of a total of 250 embryos of varying morphology between the pronucleate stage and the 8-cell stage. The overall incidence of chromosomal abnormality among these embryos was 49%. At the pronucleate stage (n = 46) the incidence was 65.2%, at the 2-4-cell stage (n = 126) it was 54.6%, and at the 5-8-cell stage (n = 78) it was 27.4%. Cleavage-stage embryos with poor morphology (irregular shaped blastomeres with severe extracellular fragmentation) showed a higher incidence of chromosomal abnormality (62%; 54 of 87 analysed) than those with good morphology (22.2%; 26 of 117 analysed). This study demonstrates: (i) that there is progressive loss of chromosomally-abnormal embryos during preimplantation development; and (ii) that there is an association between chromosomal abnormality and embryo morphology. PMID- 8726862 TI - Intracellular calcium in cultured rabbit oviduct epithelial cells. AB - Oviduct fluid is the medium in which fertilization and early embryonic development occur but little is known about the ionic basis of fluid secretion or its control. Since calcium ions (Ca2+) are involved in the mechanism of secretion in other epithelia, the intracellular calcium concentration ([Ca2+]i) was measured in single, rabbit oviduct epithelial cells in primary culture using the fluorescent dye Fura-2. The resting [Ca2+]i was constant (115 nM) in cells cultured for 2-7 days. Ion substitution experiments demonstrated the presence of a Na+/Ca(2+)-exchange system in the plasma membrane, whereas influx through channels was found to have only a minor role maintaining the resting [Ca2+]i. The addition of dibutyryl cAMP (db cAMP) induced two types of response: the first was an increase in [Ca2+]i, dependent on the presence of extracellular Ca2+, and the second was a zero response. Extracellular ATP induced a transient increase in [Ca2+]i owing to the release of Ca2+ from intracellular stores and Ca2+ entering the cell across the plasma membrane. It is proposed that these effects may be due to the presence of two types of cell in culture-the ciliated and non-ciliated (secretory type) oviduct epithelial cells. PMID- 8726863 TI - Production of extracellular matrix, fibronectin and steroidogenic enzymes, and growth of bovine granulosa cells in anchorage-independent culture. AB - A proportion of the granulosa cells from bovine antral follicles will survive, like stem cells, in anchorage-independent culture. To study these cells, bovine granulosa cells were isolated from medium-sized follicles (3-5 mm), plated out (in aliquots of 2.5 x 10(4) viable cells) onto a 1 mL agar base, and overlaid with 1 mL of methycellulose solution in culture medium (control). The cells were cultured (14 days) and then processed for histology (n = 14) or Western immunoblotting (n = 5). Under control conditions or after treatment with basic fibroblast growth factor (bFGF; 50 ng mL-1), a proportion of the granulosa cells divided to produce colonies; individual cells remained small. bFGF increased the number of cells harvested (15.8 +/- 7.3-fold, as measured indirectly by the relative amount of the nuclear La antigen), increased the average diameter of the colonies from 88.9 +/- 13.5 microns to 136.5 +/- 4.9 microns and stimulated the production of fibronectin 5.7 +/- 1.5-fold (P < 0.05). An extracellular matrix, which has previously been shown to be a basal lamina, was observed in 19.1% of the colonies (total of 350 colonies examined; n = 8 experiments). Cells treated with dibutyryl cAMP (1 mM) hypertrophied and had 50 +/- 28.7-fold and 102.6 +/- 55.8-fold higher levels of cholesterol side-chain cleavage cytochrome P450 (P < 0.001) and 3 beta-hydroxysteroid dehydrogenase (P < 0.01) respectively (n = 5). Thus, granulosa cells with characteristics of stem cells can divide and produce extracellular matrix, or be induced to differentiate when in culture without anchorage. PMID- 8726864 TI - Bovine granulosa cells express extracellular matrix proteins and their regulators during luteinization in culture. AB - This study investigated the ability of bovine granulosa cells to express and secrete collagen, metalloproteinase (MMP) activity and a tissue inhibitor of metalloproteinase (TIMP-1) during luteinization in vitro. Cells from mature (1-2 mL fluid volume) bovine follicles were cultured over 4 days in serum-free medium. Their luteinization during culture was confirmed by a 10-fold increase in progesterone secretion. Samples of cell extracts, culture media and follicular fluid were subjected to Western blotting to identify secreted proteins and to gelatin zymography to detect enzyme activity. Poly A+ RNA, isolated from cells before and after culture, was probed to detect expression of collagen alpha 1(I), collagen alpha 3(IV) and TIMP-1. The results revealed that: (1) the collagen alpha 1(I) subunit gene was expressed in cells before culture but with greater intensity by Day 4 culture; collagen I protein, on the other hand, was not detectable in culture medium; (2) the collagen alpha 3(IV) subunit gene was expressed at a low level in uncultured cells and could be detected on Day 4 of culture; low amounts of the protein were detected in medium; (3) a 92-kDa band of gelatinase activity (presumed MMP-9) was present in all medium samples, together with bands of unidentified activity; and (5) the TIMP-1 gene was expressed in uncultured cells but its expression increased markedly up to Day 4 of culture. These results show that granulosa luteinization is associated with an increase in the expression of collagen, collagen-degrading enzymes and TIMP-1. Collagen protein, however, may be only poorly synthesized in this culture model. The results suggest that granulosa-derived cells are a likely source of components of the extracellular matrix during post-ovulatory remodelling of early luteal tissue. PMID- 8726865 TI - Cryopreservation of germ cells from bovine fetal ovaries. AB - Bovine fetuses at stages required for studies of female germ cells (primordial germ cells and oogonia) become available from the abattoir at unpredictable times. To alleviate this logistical problem, a procedure to cryopreserve these ovarian germ cells has been devised. Fetal ovarian cells were dispersed and suspended in 1.5 M dimethyl sulfoxide (DMSO) prepared in modified TCM 199 medium. The suspensions were aspirated into plastic semen straws, cooled, seeded to induce ice formation at -7 degrees C, and then cooled at 1 degree C min-1 to -70 degrees C before being plunged into liquid nitrogen at -196 degrees C for storage. The straws were thawed at a moderate rate of approximately 250 degrees C min-1, the DMSO was diluted 28-fold with culture medium, and then the cells were cultured for > 2 h before their viability was tested or they were used for nuclear transfer. No statistically significant difference in viability before and after cryopreservation was detected by vital staining with fluorescein diacetate (P > 0.05). When frozen-thawed germ cells were fused to cytoplasts, the cleavage rate of the resultant reconstructed embryos 44 h after fusion was 31%, although none developed into blastocysts. It is concluded that cryopreservation of bovine fetal ovarian germ cells is feasible and can play a major role in facilitating future experimentation. PMID- 8726866 TI - Studies in sheep examining plasma follistatin elevations due to frequent blood sampling or surgery. AB - This study examined the effects of the anticoagulant heparin on plasma follistatin (FS) concentrations used during blood sampling in Corriedale ewes. Plasma FS concentrations increased acutely (P < 0.01) following a bolus intravenous injection of heparin (3200 U). To determine whether frequent sampling per se can also increase plasma FS concentrations, ewes were sampled using a substitute anticoagulant, ethylenediamine tetraacetic acid. Frequent sampling had no effect on plasma FS concentrations. alpha-2 Macroglobulin concentrations were measured before and after bilateral ovariectomy and sham ovariectomy to determine whether this agent causes an increase in plasma FS concentrations 10-12 h after laparotomy. No differences in plasma alpha-2 macroglobulin concentrations were observed at the time of the observed increase in plasma FS concentrations. IN CONCLUSION: (1) heparin caused acute elevations in plasma FS concentrations probably by binding to FS, thereby reducing its plasma clearance; (2) rapid blood sampling per se did not elevate plasma FS concentrations; (3) alpha-2 macroglobulin concentrations were not elevated at the same time as plasma FS concentrations 10-12 h after surgery; and (4) the small increase in alpha-2 macroglobulin concentrations before surgery was attributable to a stress-induced response as seen in other species. PMID- 8726867 TI - Relationships between follicle-stimulating hormone, follicle growth and ovulation rate in sheep. AB - The changes in follicle-stimulating hormone (FSH) concentration required to affect follicle growth and ovulation rate within individual ewes were examined. Relationships between peripheral FSH concentrations during the late-luteal and follicular phase and subsequent ovulation rates were investigated in 22 ewes from 4 breeds over 3 successive cycles (Experiment 1). Ewes were grouped as follows: Group 1 (n = 6), ewes exhibiting the same ovulation rate at each oestrous cycle: Group 2 (n = 5), ewes with three different ovulation rates at each oestrous cycle; and Group 3 (n = 11), ewes with the same ovulation rate at two oestrous cycles and a different ovulation rate on one occasion. Data from ewes in Group 1 and 3 provided estimates on the variation in FSH concentrations between cycles which were not large enough to alter ovulation rate (range, 0-67% variation in FSH concentration). In Group-2 ewes, there was no consistent association between increases in ovulation rate and the proportional increases in FSH concentrations. Differences in FSH concentrations were often less than those that did not alter ovulation rate in Group-I ewes. Furthermore, only 3 of 11 Group-3 ewes demonstrated high FSH concentrations associated with high ovulation rate (or low FSH concentrations and low ovulation rate) when compared with the concentrations found at the two cycles in which ovulation rate was similar. Hence, there was little evidence that FSH concentrations during the late-luteal and follicular phase are associated with changes in ovulation rate within individual ewes. In Experiment 2, follicles of similar size obtained from the same ewe (FecBFec+ and Romanov) showed markedly different responses in vitro to graded doses of FSH as measured by aromatase activity. It is concluded that, within a ewe, the large variability between gonadotrophin-dependent follicles in their requirement for FSH prevented the expression of any thresholds of ovarian response to FSH. PMID- 8726868 TI - Metacarpal growth and systemic markers of bone metabolism in the ovine fetus. AB - Bone metabolism was assessed in 56 Ile de Francex(RomanovxLimousine) fetal lambs killed between Day 80 and Day 145 of gestation. In each fetus, the length, width and weight, as well as the calcium and phosphorus content of the left diaphyseal metacarpal bone were measured. Plasma alkaline phosphatase (ALP) and tartrate resistant acid phosphatase (TRAP) activities, bone ALP (B-ALP), osteocalcin (OC) and insulin-like growth factor-I (IGF-I) were assayed in these fetuses and in six newborn lambs from birth until one month after birth. Fetal growth is characterized by an increase in bodyweight, bone size and bone mineral content from Day 80 to Day 132 of gestation. These parameters did not significantly vary until birth. Plasma concentrations of IGF-1, OC and B-ALP increased from Day 80 to Day 132. Between Day 132 and birth, plasma IGF-I and B-ALP concentrations did not significantly vary, whereas plasma OC concentration decreased, confirming the usefulness of OC as a marker of osteoblastic activity and bone formation in the ovine species. The increase in plasma IGF-I, B-ALP and OC concentrations observed during the first two weeks of postnatal life indicate an intense skeletal growth in these animals, which was confirmed by the bodyweight growth curve. PMID- 8726869 TI - Increased secondary sex ratios in golden hamster litters sired by males without coagulating glands and seminal vesicles. AB - In golden hamsters, although bilateral ablation of paternal coagulating glands (CGX) and seminal vesicles (SVX) did not affect fertility, a higher number of male pups were born. The present study aimed at determining whether this male biased sex ratio was due to an imbalance of fertilization by X and Y chromosome bearing sperms or whether it was the consequence of a sex-related differential survival of embryos. The sex of embryos sired by sham-operated (SH) controls or males subjected to bilateral ablation of ampullary glands (AGX), CGX and SVX was determined from chromosomal spreads at 10 h post coitum and 10 days post coitum. The primary sex ratio of of the SH group did not deviate from the hypothetical sex ratio of 1:1. The sex ratios of zygotes from the three experimental groups did not differ from that of the controls. However, by mid gestation, the sex ratio was significantly higher in the SVX group (P < 0.05) and the CGX group (P < 0.005). The absence of secretions from the ampullary gland, coagulating gland and seminal vesicle had no effect on the primary sex ratio, thus these glands did not appear to affect fertilization by the X and Y chromosome-bearing sperm. The increased secondary sex ratios observed in the SVX and CGX groups were due to the preferential survival of males. PMID- 8726870 TI - Effects of nitric oxide synthase inhibitors on the outcome of in vitro fertilization in the mouse. AB - The effect of three nitric oxide (NO) synthase inhibitors, L-NG-nitro-arginine (NO2Arg), NG-Nitro-L-arginine methyl ester (L-NAME) and aminoguanidine, on in vitro fertilization in the mouse was examined. Mouse epididymal spermatozoa were capacitated in a medium with or without NO synthase inhibitors. Oocytes were inseminated and the percentage of oocytes with two pronuclei was scored after an 8-h incubation. NO2Arg and L-NAME, but not aminoguanidine, inhibited fertilization. L-NAME inhibited fertilization in a dose-dependent manner, and its effects were stereospecific. The inhibitory effect was neutralized by L-arginine but not by D-arginine. Moreover, D-NAME did not inhibit fertilization. The results suggest that NO synthase activity (presumably of the constitutive type is necessary for spermatozoa to display their full fertilizing ability. PMID- 8726871 TI - Cloning and sequencing of a cDNA encoding an ovine oestrus-associated oviducal protein. AB - Ovine oestrus-associated oviducal glycoprotein (oEGP) is synthesized and secreted specifically by the ampullary region of the ovine oviduct during the peri ovulatory stages of the oestrous cycle. A cDNA that encodes oEGP was isolated and sequenced. Isolation of oEGP was achieved using the polymerase chain reaction (PCR) with primers based on a bovine oestrus-associated oviducal glycoprotein cDNA (bOGP) sequence. A 1599-bp cDNA encodes, in part, a deduced 519-amino acid sequence of mature protein which carries two potential N-linked glycosylation sites. The deduced amino acid sequence is more than 95% identical to that of bOGP and more than 74% identical to the first 491 amino acids of human oestrogen dependent oviducal glycoprotein (hOGP). Northern blot hybridizations of RNA from several sheep tissues detected mRNA (2.4 kb) only in an ampulla oviduct sample. PMID- 8726872 TI - Ultrasound-related litigation in obstetrics and gynecology: the need for defensive scanning. PMID- 8726873 TI - Biochemical and ultrasound screening for Down's syndrome: rivals or partners? PMID- 8726874 TI - Fetal heart rate in trisomy 21 and other chromosomal abnormalities at 10-14 weeks of gestation. AB - Fetal heart rate was measured routinely as part of a prospective study examining the efficacy of screening for trisomy 21 by fetal nuchal translucency thickness and maternal age. In 6903 normal singleton pregnancies the fetal heart rate decreased from a mean of 171 bpm at 10 weeks of gestation to 156 bpm at 14 weeks (r = 0.413, p < 0.0001). In 85 trisomy 21 pregnancies, the mean heart rate was significantly higher than in the normal group (mean difference 0.67 SD, 95% confidence interval 0.42-0.92, t = 5.3, p < 0.001). The fetal heart rate in trisomy 18 and triploid fetuses was significantly lower and in trisomy 13 and Turner syndrome was higher than normal. There was no significant association between delta fetal heart rate and delta nuchal translucency thickness in either the normal (r = -0.018) or the trisomy 21 (r = -0.031) pregnancies. Consequently, the risk for chromosomal defects can be derived by combining data from maternal age, fetal nuchal translucency and fetal heart rate. The effectiveness of screening by this method was examined in a self-selected population with completed pregnancies that had undergone first-trimester scanning. This population contained 6903 normal and 29 trisomy 21 fetuses. For a false-positive rate of about 5%, the sensitivity for trisomy 21 was 48% by maternal age, 26% by fetal heart rate, 72% by nuchal translucency thickness, 59% by maternal age and fetal heart rate, 76% by maternal age and nuchal translucency thickness and 83% by a combination of maternal age, nuchal translucency thickness and fetal heart rate. PMID- 8726875 TI - Abnormalities of the heart and great arteries in chromosomally normal fetuses with increased nuchal translucency thickness at 11-13 weeks of gestation. AB - Pathological examination of trisomic fetuses with increased nuchal translucency thickness at 11-13 weeks of gestation demonstrated a high prevalence of cardiac defects and abnormalities of the great arteries. This study reports the pathological findings observed from the examination of the heart and great arteries of 21 chromosomally normal fetuses with increased nuchal translucency. In 19 of the 21 cases there were abnormalities; the commonest was narrowing of the aorta at the level of the isthmus and immediately above the aortic valve. This finding is different from that in case of trisomy 21, where narrowing of the isthmus is associated with an increased diameter of the aortic valve. These findings suggest that abnormalities of the heart and great arteries may be implicated in the pathogenesis of increased nuchal translucency not only in trisomic fetuses but also in chromosomally normal fetuses. It can be implied that increased nuchal translucency thickness at 10-14 weeks of gestation may prove to be a useful marker for the identification of fetal cardiac abnormalities. PMID- 8726876 TI - Prenatal ultrasonographic diagnosis of fetal hepatic hyperechogenicities: clinical significance and implications for management. AB - The clinical significance of sonographically detected fetal hepatic hyperechogenicities has not been fully established. The aim of this study was to assess various aspects of fetal hepatic hyperechogenicities detected in utero, including natural history, prenatal investigation and pregnancy outcome. In a retrospective survey of pregnant women presenting to the ultrasonographic unit, five fetuses with a mean gestational age of 20 weeks (range 14-24 weeks) were antenatally diagnosed as having hepatic hyperechogenicities. All cases underwent extensive investigation to establish the etiology and to determine the spectrum of ultrasonographic presentation. In three fetuses, only parenchymal lesions were found, while two had combined lesions: parenchymal and peritoneal. The earliest prenatal ultrasonographic diagnosis was made at 14 weeks' gestation in a fetus with combined hyperechogenic lesions, which, at 12 weeks' gestation, demonstrated unexplained fetal ascites. Associated abnormalities were found in only one fetus in which a solitary parenchymal lesion was associated with direct communication of the intrahepatic portion of the umbilical vein with the right atrium, and high output cardiac failure. In this case, termination of pregnancy confirmed ischemic hepatic necrosis. In the other four cases, no direct etiology could be determined, and all four pregnancies continued to term with normal neonatal follow up until 12 months of age. The present data suggest that in cases of fetal hepatic hyperechogenicities, where apparent intrauterine infection or neoplasia can be ruled out, a vascular disruption phenomenon is a putative etiology. If there are no associated morphological abnormalities or abnormal karyotypes, the prognosis may be favorable. PMID- 8726877 TI - Fetal echocardiography guidelines to predict survival of fetuses with ascites. AB - In a retrospective analysis of the standard protocols for fetal echocardiographic examination, 27 fetuses (mean gestational age 29 +/- 5.4 weeks) with ascites were evaluated. Fetal cardiomegaly (increased heart area/chest area ratio), the presence or absence of atrioventricular valve regurgitation, inferior vena cava and ductus venosus Doppler flow velocity and umbilical vein pulsation were evaluated quantitatively in a group of survivors and non-survivors. A statistically significant difference between the two groups was found for the presence of atrioventricular valve regurgitation (p = 0.003), and for cardiomegaly (p = 0.009). There was no statistical difference for the presence of abnormal venous flow and umbilical pulsation (p > 0.05). Abnormal venous Doppler flow velocities in the inferior vena cava and ductus venosus were observed more frequently than umbilical vein pulsation. The mean heart area/chest area ratios in the group of survivors and in the group of non-survivors were 0.34 +/- 0.09 and 0.48 +/- 0.08, respectively (p < 0.001). The mean gestational ages at the time of diagnosis of ascites in the groups of survivors and non-survivors were 33 +/- 3.3 weeks and 28 +/- 5.1 weeks, respectively (p < 0.05); the mean gestational ages at the time of delivery were 35.6 +/- 2.3 weeks and 33.3 +/- 4.9 weeks, respectively (p = 0.33). In terms of different echocardiographic features as well as the gestational age of the unborn patient with ascites, there is apparently no single indicator of a poor prognosis. Our results suggest that the prognosis is usually poor in a fetus with ascites, when cardiomegaly is detected, as well as the presence of tricuspid and mitral valve regurgitation. This is regardless of the type of venous flow and regardless of the etiology of the ascites. PMID- 8726878 TI - Venous Doppler in the fetus with absent end-diastolic flow in the umbilical artery. AB - High perinatal mortality has been reported in association with the finding of absent end-diastolic flow velocities in the umbilical artery. The fetus is known to centralize its circulation during hypoxemia and abnormal venous blood flow velocities have been reported in cases of heart failure and imminent asphyxia. The aim of this study was to evaluate blood flow velocities recorded with Doppler ultrasound in the umbilical vein, inferior vena cava and middle cerebral artery as predictors of survival in 17 fetuses with absent or reversed end-diastolic flow in the umbilical artery. There were five perinatal deaths, all having abnormal umbilical cord venous pulsations. An increased proportion of diastolic blood flow velocity in the middle cerebral artery as a sign of hypoxia was found in 12 fetuses without a relationship to perinatal mortality. An increased proportion of retrograde blood flow in the inferior vena cava was recorded in only one fetus, which died on the same day of congestive heart failure, suggesting fetal heart sparing in the remaining complicated pregnancies. The results suggest that abnormal end-diastolic umbilical venous pulsation in the cord is a late and ominous sign of a severely compromised fetus, while abnormal blood flow velocimetry in the middle cerebral artery might be an earlier sign of fetal hypoxia, with a better prognosis. PMID- 8726879 TI - Fetal urine production estimated with ultrasound. The lower limit of normality is illustrated in a case with severe hypoplasia of the kidneys. AB - Urine production and weight deviation in a growth-retarded fetus with oligohydramnios was estimated by ultrasound on four occasions between weeks 27 and 33 of gestational age. The fetus demonstrated facial features typical of Potter's syndrome. Although this syndrome is associated with bilateral renal agenesis, the fetus in the present case presented with hypoplastic kidneys. Urine production was reduced when measured on three occasions but was surprisingly normal at week 31. In this report, we describe a way to assess urine production in a fetus. PMID- 8726880 TI - Estimation of fetal shoulder width by measurement of the humerospinous distance by ultrasound. AB - The present study was undertaken to find a practical method for estimating the fetal shoulder width by ultrasound. Sixty pregnant women at term were included in the study. The distance between the lateral margin of the fetal cartilaginous caput humeri and the processus spinosus of the cervical vertebra C7 (humerospinous distance) was measured by ultrasound. The neonatal shoulder width was measured with a craniometer during the first postnatal day. Linear regression analysis was used to calculate the correlation between the humerospinous distance and the shoulder width, and a significant linear correlation (r = 0.612, p < 0.001) was shown. Although, in this small study, we could not show a relationship between humerospinous distance and dystocia, we feel a clinical study of humerospinous distance as a predictor of shoulder dystocia is indicated. PMID- 8726881 TI - Pathognomonic echo patterns of benign cystic teratomas of the ovary: classification, incidence and accuracy rate of sonographic diagnosis. AB - The great variability in the sonographic appearance of ovarian cystic teratomas poses difficulties for their diagnosis and classification. To overcome such difficulties, we have proposed a simplified classification of the pathognomonic echo patterns of ovarian cystic teratomas based on three basic types of echo pattern. In a prospective study of 118 echogenic adnexal masses, with postoperative histological confirmation, this classification enabled the correct diagnosis to be made in 115 cases (accuracy rate, 97.45%). In the three cases misdiagnosed preoperatively as ovarian cystic teratoma, the tumors proved to be benign and included a serous cyst adenoma, a serous cyst adenofibroma and a Brenner tumor. We suggest that, with our proposed classification of the echo patterns of ovarian cystic teratomas, sonography can become a quick and accurate tool in the recognition of this condition. PMID- 8726882 TI - Role of transvaginal ultrasound color flow imaging and Doppler waveform analysis in differentiating between benign and malignant ovarian tumors. AB - This study was undertaken to assess the usefulness of ultrasound color flow imaging and Doppler waveform analysis in differentiating between benign and malignant ovarian tumors. From 1991 to 1993, 146 patients with 156 ovarian tumors underwent transvaginal scanning with color flow imaging and Doppler waveform analysis prior to surgery. All the ovarian tumors were removed surgically and confirmed histologically. The sensitivity and specificity of color flow imaging in identifying malignant ovarian tumors was 76.5% and 68%, respectively. Positive and negative predictive values were 40% and 91.2%, respectively. The addition of Doppler waveform analysis failed to produce statistically better results. Seven of the eight cases of ovarian cancer were missed by color Doppler. There were no solid areas present and malignancy was not suspected. Both modalities also failed to identify any case of ovarian cancer presenting as a simple cyst. PMID- 8726883 TI - Sonographic follow-up of a placenta left in situ after delivery of the fetus in an abdominal pregnancy. AB - A case of abdominal pregnancy was followed until 34 weeks' gestation, when rupture of the amniotic sac was noted. A live and anatomically normal female weighing 1400 g was delivered by laparotomy and the placenta was left in place. Color Doppler imaging and measurement of serum beta-human chorionic gonadotropin (beta-hCG) were successfully used to follow the placental involution after delivery. A progressive increase in the resistance index in the utero-ovarian and subplacental vessels was observed while beta-hCG disappeared 45 days after laparotomy. The mother and baby left hospital 20 days after delivery and are both doing well. PMID- 8726884 TI - Antenatal diagnosis of a late abdominal pregnancy using ultrasound and magnetic resonance imaging: a case report of successful outcome. AB - A case of an advanced abdominal pregnancy diagnosed by ultrasound and magnetic resonance imaging (MRI) at 21 weeks' gestation is presented. The complementary roles of ultrasound and MRI in achieving accurate diagnosis and the role of ultrasound in ongoing pregnancy assessment are emphasized. PMID- 8726885 TI - The significance of echogenic areas in the fetal abdomen. PMID- 8726886 TI - Abnormal fetal pulmonary venous blood flow velocity waveforms in the presence of complete transposition of the great arteries. PMID- 8726887 TI - Searching for the Magic Johnson effect: AIDS, adolescents, and celebrity disclosure. AB - The objectives of this study were to measure changes in AIDS-related attitudes and behaviors in adolescents in the 13 months following Magic Johnson's disclosure that he was HIV positive, and to test whether gender, race, age, sexual experience, and pre-existing HIV-avoidant behaviors would emerge as significant dependent variables. Adolescent clinic attendees (N = 181) ages 12-19 in four cities completed a questionnaire assessing change in AIDS-related attitudes and behaviors since Johnson's announcement. Respondents were divided into low-risk and at-risk groups. Sixty percent of respondents reported that Magic Johnson's announcement had increased their awareness of AIDS, 65.4% reported increased self-efficacy in a sexual situation, 37.2% reported that they had changed their perceived AIDS risk, 37.8% described increased resistance to peer pressure for sexual intercourse. The low-risk group was more likely to report increased self-efficacy and resistance to peer pressure but no change in perceived risk or increased AIDS awareness. Significant relationships were found between gender and increased AIDS awareness, gender and increased resistance to peer pressure to engage in sexual intercourse, race and increased AIDS awareness, and more lifetime sex partners and increased self-efficacy. PMID- 8726888 TI - Lifestyle and risk of premature sexual activity in a high school population of Seventh-Day Adventists: Valuegenesis 1989. AB - In the past 20 years, sexually transmitted diseases (STDs), including AIDS, and the physical, psychological, and economic difficulties associated with unwanted pregnancy have increased steadily among American adolescents. The objective of this study was to evaluate Adventist lifestyle as a modification of popular American culture which reduces the risk of early sexual activity in adolescents and thus also reduces the risk for both STDs and teen pregnancy. The study was based on 8,321 respondents to a questionnaire concerning specific behaviors, beliefs, and attitudes among Seventh-Day Adventist youth attending 58 high schools in North American. Analysis of the data demonstrated that a wide variety of behaviors were associated with premature sexual activity, including previously reported high-risk behaviors such as drug or alcohol use. In addition, several behaviors that are discouraged within Adventist culture, such as going to a movie theater or participating in competitive sports, also were associated with early sexual activity. It is hypothesized that these latter behaviors may predict the emergence of other high-risk behaviors, such as early sexual activity, in both Adventist and popular cultures, and thus may be "transition-marking behaviors" as described by Jessor and Jessor (1975). PMID- 8726889 TI - The source, accuracy, and impact of initial sexuality information on lifetime wellness. AB - Seven hundred male and female respondents ranging in age from 9 to 73 years, were surveyed. Data were provided for examining the age, source, accuracy, and impact of initial sexuality information on lifetime wellness. Subjects were placed in five chronological age groups for data analysis. Responses from the youngest group, comprised exclusively of students enrolled in schools offering comprehensive school health education, were also utilized to examine program influence. While the age for obtaining sexual knowledge was similar, parents were never reported as the primary information source. Young participants, citing teachers and siblings as their primary information source, achieved the highest accuracy scores. This suggests the value of health education. Accurate sexuality information was reported as extremely important to lifetime wellness, and ignorance was found to have produced guilt and illness in the majority of respondents. PMID- 8726890 TI - An empirical investigation of the construct of personality authority in late adolescent women and their level of college adjustment. AB - A random sample of 102 women students from a small private college were assessed concerning their levels of individuation, intimacy, intimidation, parental triangulation, and personal authority. These concepts formed the construct of Personal Authority and were tested as to their relationship to self-esteem, physical health, and college adjustment. Results generally supported better adjustment for those women who had higher levels of Personal Authority. PMID- 8726891 TI - The influence of peer affiliation and student activities on adolescent drug involvement. AB - This study examined the importance of students' academic performance level and extracurricular activities as predictors of drug involvement relative to peer influence. Social development theory provided the theoretical rational for the study. Data were obtained from 2,229 randomly selected students in the eighth, tenth, and twelfth grades from seventeen school districts in northeastern Ohio. At all three grade levels, involvement in extracurricular activities and academic level were significantly correlated with students' gateway and hard drug use. Consistent with prior research, the strongest correlate of gateway and hard drug use across all grade levels was affiliation with drug-using friends. Having a job after school was marginally related to self-reported gateway drug use at grade level ten. Multiple regression analysis revealed that extracurricular involvement and academic performance level make small, but unique contributions to the prediction of adolescents' gateway drug use beyond affiliation with drug-using peers at all three grade levels. The findings of this study suggest that students' academic performance and extracurricular involvements are significantly related to adolescent gateway and hard drug use, but have less predictive significance relative to peer relationships. PMID- 8726892 TI - Effects of family structure on the adolescent separation-individuation process. AB - This study examined the effects of college students' family structures on the separation-individuation process. Family structure groups investigated were (1) intact; (2) divorced, mother-custody, no remarriage; and (3) divorced, mother custody, remarried. The components of the separation-individuation process examined were attachment to parents, conflictual independence, internal self awareness, ego identity, and self-esteem. Results indicated that adolescents from intact families perceived themselves as having more conflictual independence from and more positive emotional relationships with their fathers than did those from divorced or remarried families. The groups did not differ in ego identity or self awareness. Both attachment and conflictual independence from fathers were related to self-esteem. Conflictual independence from mothers was also related to self esteem, while attachment to mothers was not. Ego identity was related to self esteem, but self-awareness was not. The discussion addresses both clinical and future research implications. PMID- 8726893 TI - A critique of social bonding and control theory of delinquency using the principles of Psychology of Mind. AB - The present paper first describes the refined principles of Psychology of Mind (POM) and shows how their logical interaction can help explain the comparative amounts of both deviant and conforming behavior engaged in by youthful offenders. The logic of these principles is then used to examine the major assumptions of social bonding and control theory of delinquency focusing predominantly on the formulations of Walter Reckless (1956) and Travis Hirschi (1969). PMID- 8726894 TI - Predictors of adolescent A. A. affiliation. AB - Alcoholics who receive treatment in in-patient settings are routinely referred to Alcoholics Anonymous upon discharge, yet not all affiliate with A.A. The characteristics of A.A. affiliators have been explored in the past to further improve discharge planning, but to date no studies have described the characteristics of adolescents who affiliate with A.A. The sample used in this study was 70 adolescents who had completed in-patient treatment and were contacted as part of a follow-up survey. Half of the group had affiliated with A.A. A discriminant analysis was used to predict affiliation, and the study found that affiliators were more likely to have had prior treatment, had friends who did not use drugs, had less parental involvement while in treatment, and more feelings of hopelessness. Possible explanations for these findings are discussed as well as areas for further research. PMID- 8726895 TI - Adolescent homicide and family pathology: implications for research and treatment with adolescents. AB - The origins of serious violence in adolescents has been of concern throughout the century. The number of felony-murder convictions of adolescents on death row in the United States is arguably a warning beacon about the serious nature of juvenile homicide. This article reviews the research on adolescent homicide and highlights significant family variables. The contributions of a family studies perspective for understanding adolescent homicide is discussed and implications suggested. PMID- 8726896 TI - Patterns of parenting during adolescence: perceptions of adolescents and parents. AB - The purposes of this study were (1) to explore differences in maternal and paternal parenting style and parental involvement, (2) to examine the differences between parents' and adolescents' perceptions of parenting style and parental involvement, and (3) to explore the changes in parenting style and parental involvement between the adolescents' ninth and twelfth grade years. Subjects were 244 ninth graders recruited from several school districts in the Southeast and Midwest. Thirty-one subjects participated in a small longitudinal study three years later when they were in the twelfth grade. Measures of maternal and parental demandingness, responsiveness, values toward achievement, involvement in schoolwork, and involvement in school functions, designed for this program of research, were obtained from both adolescents and their parents. Results showed that both adolescents and parents perceived mothers to be more involved in parenting than were fathers during both ninth and twelfth grades. Mothers and fathers were not found to be different regarding their values toward achievement. Additionally, both mothers and fathers perceived themselves to be higher on all aspects of parenting than their adolescents perceived them to be during both ninth and twelfth grades. In the longitudinal study, both adolescents and parents perceived levels of parenting to drop between ninth and twelfth grades, except values toward achievement which did not change. Implications for interpretation of existing research and for considerations of future research are discussed. PMID- 8726897 TI - Cognitive-affective sources of sport enjoyment in adolescent sport participants. AB - Cognitive-affective sources of sport enjoyment were examined among male high school freshmen (N = 231), all of whom had participated in organized youth sport. Subjects completed several sport-specific measures. Also, recorded was an indicator of number of years of participation in organized sport. Stepwise multiple regression analysis indicated that significant sources of sport enjoyment included task orientation, perceived competence, learned helpless affect, and years of participation in organized sport. Results also indicated, however, that ego orientation failed to enter the stepwise equation as a significant predictor of sport enjoyment. The findings are discussed in regard to the importance of the development of competence to enjoyment. Suggestions are offered for the need to advance task orientation and self-referenced perceived ability in sport achievement settings in order to foster positive affect and ongoing participation in organized youth sport. PMID- 8726898 TI - Measurement of ethical behavior in leisure among high- and low-risk adolescents. AB - Based on Aristotelian ethics, an "Adolescent Ethical Behavior in Leisure Scale" (AEBLS) was developed. Sixty-two items were created and reviewed by a panel of experts for content-related evidence of validity. Two validity studies were then conducted. In Study 1, data from 419 high- and low-risk adolescents were used to examine correlations between the AEBLS scores and measures of substance use, school bonding, and dispositional leisure boredom. All hypotheses about relations between these variables and the AEBLS were supported. In Study 2, an experiment was conducted to assess construct-related evidence of validity. Seventy-one adolescents completed the AEBLS and evaluated images of sensory and cortical recreation activities to which they were exposed. Consistent with the hypotheses, significant positive correlations were found between the AEBLS and evaluation of cortical activities, and significant negative correlations were found between the AEBLS and evaluation of sensory activities. Results support the use of AEBLS scores to discriminate between ethical leisure behaviors of high- and low-risk adolescents. PMID- 8726899 TI - The long-term impact of AIDS-preventive interventions for delinquent and abused adolescents. AB - This study examines whether cognitive/behavioral interventions that produced immediate changes in AIDS-related knowledge, attitudes, and intentions for coping with AIDS-risk situations among delinquents and abused adolescents, are capable of producing long-term benefits assessed at 9-12 months follow-up. Adolescents (N = 218) from 15 residential centers received an intensive nine-session HIV prevention program. Centers were randomly assigned to skills training, discussion only, or control groups. Results showed that one intervention model, discussion groups, produced a long-term increase in knowledge about AIDS and higher reported intentions to cope with AIDS-risk situations. However, both skills-training and discussion groups did not produce a long-term reduction in the level of engagement in high-risk behaviors. Several reasons for these results are discussed, with emphasis on an explanation based on the theoretical perspective of life chances or life options. PMID- 8726900 TI - Street children in South Africa: findings from interviews on the background of street children in Pretoria, South Africa. AB - In the present study findings from interviews on the background of street children in Pretoria, South Africa are presented, discussed, and compared with research done in the past on South African street children. Findings from the investigation indicate that the average age of South African street children is approximately 13 years, predominantly of male African origin. Most have been on the streets for three years or longer, and they cited family violence, parental alcoholism, abuse, and poverty as the main motivating factors for leaving home. Most of these findings are common among street children all over the world. PMID- 8726901 TI - Adolescents' perceptions of family functioning in the United Arab Emirates. AB - A sample of 710 subjects between 14 and 19 years of age were examined to assess psychological health of United Arab Emirates (UAE) families with adolescent members. The Family Functioning Questionnaire (FFQ) based upon the adolescent's perception of his or her family life and climate contains seven dimensions that constitute the family functioning process: family structure, emotional fulfillment, internal relations, behavioral control, value transmission, basic requirements, and external relations. Results indicate that the adolescents perceived their families to be operating more toward the functional than dysfunctional end of the scale. Males scored their families more functional in family structure and in external relations more significantly than did females. Females perceived their families as more functioning in internal relations, behavioral control, and basic requirements. However, a trend was found in favor of males on emotional fulfillment. Family functioning scores were higher in upper social economic status (SES) families than in other SES levels. PMID- 8726902 TI - Biopsychopathologic risk profile of adolescents with eating disorder symptoms. AB - The aim of this study is to determine whether there is a psychological and somatic pattern for adolescents with eating symptomatology. The Eating Attitudes Test (Garner & Garfinkel, 1979) and other psychological instruments to evaluate self-esteem (SEI, Battle, 1981), personality (EPQ-J, Eysenck & Eysenck, 1984), school abilities (AAT, Thurstone & Thurstone, 1986), anxiety (STAIC, Spielberger, 1973) and symptoms of depression (CDI, Kovacs, 1983) were used in a sample of 515 adolescents between 13 and 14 years of age. Prevalence of risky eating attitudes was 12.4% for the girls and 8.3% for the boys. In general, a low level of self esteem and a high level of anxiety were the most important predictors of eating symtomatology, but there were differences between genders. Girls with eating symptomatology exhibited a profile with more psychopathological traits. Heavy and corpulent subjects, boys or girls, form a group with a high risk of eating disorder. This suggests that the conjunction of psychological and somatic factors at the beginning of adolescence can be a useful marker for early intervention. PMID- 8726903 TI - Determinants of suicide ideation: a comparison of Chinese and American college students. AB - A LISREL model that incorporates both social and psychological factors was used to explain Chinese and American college students' suicide ideation. Questionnaire data were obtained from one Chinese sample (N = 320) from four universities in Beijing and one American sample (N = 452) from one university in the Rocky Mountain area. As in the American sample, Chinese females score higher on the ideation scale than Chinese males, but the overall rate is lower for the Chinese than for the American college students. The findings in the American data support previous literature that family cohesion and religiosity are inversely related to suicide ideation, while the Chinese data suggest a positive correlation between religiosity and suicide ideation. This article offers a comparison of different cultural environments for Chinese and American adolescent development. PMID- 8726904 TI - Hospital-based behavior modification program for adolescents: evaluation and predictors of outcome. AB - This study is an evaluation of a residential short-term program (10-12 weeks) which utilizes a behavior modification program (BMP) in an inpatient unit for adolescents with mostly conduct problems. Evaluation included predictors of outcome such as age, gender, diagnosis, length of stay, father's presence, other treatments received, and presence of learning problems. The design used was a retrospective analysis of performance of patients on BMP measured by mean point earned in the first four weeks (pre-test) and the last 4 weeks of admission (post test). Subjects were 60 adolescents aged 12-18 enrolled for at least eight weeks in the BMP. Findings showed that the BMP was generally beneficial especially for females and nonconduct-disordered patients. Factors such as father's presence, and absence of learning problems were associated with better outcome. Age and length of stay were not correlated with significant improvement. It was found that the BMP was effective in reducing frequency of undesirable behaviors for the majority of adolescents with conduct problems. However, males with a diagnosis of conduct disorder might require a longer stay for the program to be effective. PMID- 8726905 TI - Acculturative stress among Amerasian refugees: gender and racial differences. AB - Adjustment and resettlement of refugees from one culture to another is often a challenging and sometimes difficult process. Since Congress passed the Amerasian Homecoming Act in 1987, there has been a substantial increase in the number of immigrants born of American servicemen and Vietnamese women during the Vietnam War. Several thousand Amerasians and their families left Vietnam and Cambodia for resettlement, and there are currently over thirty cluster sites of voluntary organizations used for placement and resettlement. One area is Springfield, Massachusetts, where our study is based. This research explored the mental health, adjustment, and issues of acculturation among refugees of this subgroup as they attempted to adapt to the American culture. A refugee acculturative stress inventory modified to fit this group was utilized, and Pearson correlation and t-test were employed for statistical analysis. Major findings indicate that most newly arrived Amerasians experience acculturative stress primarily in areas of spoken English, employment, and limited formal education. Gender and race had no impact on acculturative stress. Findings did not support the literature on refugees which indicates that males are more at risk than females regarding acculturative stress. There was a significant correlation between effective spoken English and employment on the level of stress. Implications for social work practice and future research are suggested. PMID- 8726906 TI - Exposure to violence and post-traumatic stress disorder in urban adolescents. AB - The prevalence and severity of stressors in the lives of urban adolescents may predispose them to symptoms of psychological stress and Post-Traumatic Stress Disorder (PTSD). The purpose of this study was to investigate variables associated with the incidence of symptomatology characteristic of Post-Traumatic Stress Disorder in adolescents in a major metropolitan area. One hundred and three high school juniors completed The Keane PTSD Scale, the Civilian Mississippi Scale for PTSD, and a demographic questionnaire was completed by 97 high school juniors of whom 29% indicated clinical levels of PTSD symptomatology. Comparisons of incidence were made with regard to gender, ethnicity, family constellation, self-reported exposure to violence, self-reported exposure to trauma, and incidence of violent crime in the vicinity of school attendance. Results revealed that minority males were exposed in their neighborhoods and schools to more violent crime than were any other group. A gender effect in response to violent crime was evident. In the school with the highest rates of murder, assault, and individual robbery, boys achieved low to mid-range PTSD scores, while girls obtained the highest PTSD scores of all participants in the study, indicating that girls may respond to violence with more symptoms of PTSD than boys. PMID- 8726907 TI - Temporal bone imaging. AB - Ameliorated computed tomography techniques and new magnetic resonance sequences have led to an important improvement in temporal bone imaging. Computed tomography is still the method of choice for imaging of temporal bone fractures, middle ear disease, and conductive hearing loss, although magnetic imaging can add important information. Patients with lesions of the cerebellopontine angle; internal auditory canal; inner ear; and, in general, all patients with sensorineural hearing loss, vertigo, and tinnitus are best examined with magnetic resonance imaging. In some cases, however, such as congenital malformations and petrous apex lesions, magnetic resonance imaging and computed tomography are complementary. The value of both modalities is discussed. PMID- 8726909 TI - Sinonasal imaging. AB - Recent changes in sinonasal imaging are a direct result of the development of functional endoscopic sinus surgery. Because of this technique, radiologists have noted an increased volume of sinus imaging, developed new imaging techniques, and are interpreting films in a different manner. This article covers the common variants seen on coronal computed tomography, discusses the complications of functional endoscopic sinus surgery, reviews the radiographic criteria for sinusitis, and addresses the role of computed tomography and magnetic resonance imaging in evaluating the sinonasal cavity. PMID- 8726908 TI - Ocular and orbital imaging. AB - The development of magnetic resonance imaging technology has proven to be a great breakthrough in diagnostic medical imaging, ophthalmologic imaging, and biomedical research. The anatomic detail demonstrated by an magnetic resonance image is a representation of at least three physical properties of static tissue: photon density and T1 and T2 relaxation times. Intrinsic differences in proton density and proton relaxation times of tissues allow excellent image contrast between various normal structures and high sensitivity for detecting pathologic states. This article discusses magnetic resonance imaging features of common ocular and orbital lesions, with emphasis on the potential of magnetic resonance imaging in ophthalmology. PMID- 8726910 TI - Temporomandibular joint and dental imaging. AB - CT scanning of the jaws with cross-sectional images perpendicular to the curvature of the alveolar processes can be used to facilitate surgical planning for placement of dental implants. After ruling out pathologic changes, the radiographic evaluation concentrates on the dimensions of the alveolar processes relative to adjacent structures. PMID- 8726911 TI - Base of skull, nasopharynx, and parapharyngeal space. AB - The base of skull, the nasopharynx, and the parapharyngeal space are among the most anatomically complex regions of the body. When pathologic changes arise in these areas, the close topographic relationship of soft-tissue structures and central nervous system structures, often separated only by an osseous or a thin soft tissue layer, generally necessitates sensitive diagnostic and complex therapeutic procedures. PMID- 8726912 TI - Oral cavity, oropharynx, and salivary glands. AB - In the oral cavity and oropharynx, the predominant disease is cancer, which arises, in most cases, from the oral mucosa. Therefore, radiologists have a minor role in the detection of the disease but help to define the exact extent of the lesion and have an important role in therapeutic planning. Computed tomography and magnetic resonance imaging are the imaging modalities of choice. In the salivary glands, nonmodalities are numerous and include computed tomography, magnetic resonance imaging, sialography, sonography, and scintigraphy. The role of radiologists is to select the best study to determine the nature of the lesion and its extent and consequences. The choice of first study is primarily based on physical examination. PMID- 8726913 TI - Imaging of the larynx. AB - CT and MR imaging are the main modalities for examination of laryngeal pathology. In general, MR imaging seems to be the optimal method of examination in cooperative patients, especially for evaluation of their larynx prior to an attempted partial laryngectomy. CT is recommended in patients who may have rapid breathing or coughing or if MR imaging is contraindicated. The choice between the two modalities is also determined by the experience of the radiologist with these modalities. Both techniques are comparable in delineating site and extent of pathology in fat and muscular tissue. MR imaging is more sensitive than CT in detecting pathologic involvement of the cartilages. CT and MR imaging are helpful in characterization of cartilaginous tumors and benign lesions, such as laryngoceles and cysts. CT is used to assess the integrity of the laryngeal skeleton in patients who suffered from trauma, that is, for identification of occult fractures, dislocations of cartilages, or confirmation of suspected laryngeal injuries. PMID- 8726914 TI - Imaging of cervical lymphadenopathy. AB - Imaging of the neck is important to diagnose occult lesions and stage the neck in cancer patients and, to a lesser extent, help differentiate swellings in the neck. Furthermore, extension of tumors can be depicted, although assessment of operability is not very reliable. Conventional imaging techniques, such as CT, MR imaging, and US, are rapidly evolving by improvement of spatial resolution and contrast, reduction of artifacts, and development of new contrast agents. Unreliable criteria for metastases, such as size and shape, will be replaced in the future by the depiction of the microstructure inside lymph nodes and the development of specific contrast agents. US-guided aspiration is currently one of the most accurate techniques to assess occult metastases as it overcomes the criteria used by CT, MR imaging, or US without aspiration. It should be noted, however, that approximately 25% of electively operated sides of the neck contain exclusively metastases smaller than 3 mm, limiting the sensitivity in these necks to 75% for all imaging modalities. New imaging modalities, such as immunoimaging with SPECT, thallium SPECT, PET, and fused images, are rapidly developing. Although these techniques will probably become very accurate for the staging of the neck, it is doubtful whether they will be used routinely to stage the neck because of the issues of cost and availability. Their role will probably be more in the detection of unknown primaries, distant metastases, and follow-up, after radiotherapy in case of clinical doubt. PMID- 8726915 TI - Parathyroid and thyroid imaging. AB - Whether to perform preoperative imaging for parathyroid adenomas is controversial in the "virgin neck" because surgery without imaging is successful in more than 90% of cases. To detect ectopic adenomas or unresected adenomas after failure of initial surgery often requires both a functional study, such as technetium-99m sestamibi nuclear scintigram, and a morphologic study, such as ultrasonography, computed tomography, or magnetic resonance imaging for anatomic landmarks. Most dominant thyroid masses require fine-needle aspiration for diagnosis. Nuclear scintigraphy is useful when the lesion is warm or hot, making a malignancy less likely, but cold lesions are nonspecific. Unless a mass is shown to infiltrate the surrounding soft tissue or spread to lymph nodes, the findings on most cross sectional imaging studies are also nonspecific. PMID- 8726916 TI - Interventional magnetic resonance imaging of the head and neck and new imaging techniques. AB - The use of magnetic resonance imaging to directly guide therapy is a new area of investigation. Clinical magnetic resonance imaging has the potential to evolve from a purely diagnostic tool into a guide for invasive procedures. The idea of using magnetic resonance to guide percutaneous biopsy developed one decade ago. Magnetic resonance-guided fine-needle aspiration for head and neck lesions has become a standard technique some institutions. Once the needle reaches a lesion in the head and neck, it is natural to explore the possibility of treatment in the same setting. The concept of interventional magnetic resonance imaging has generated interest among radiologists, surgeons, and manufacturers. This article provides an overview of interventional procedures of the head and neck and other new imaging techniques that have been developed. PMID- 8726917 TI - Interventional neuroradiology of the head and neck. AB - Interventional neuroradiologic techniques continue to assume increasing importance in the management of disorders of the head and neck. Their usefulness includes not only treatment for neoplastic and vascular conditions with embolization but also test occlusion for diagnostic information. Future developments also may extend the usefulness of interventional techniques to additional conditions, including prevention and treatment for ischemic stroke. PMID- 8726918 TI - The role of computed tomography and magnetic resonance imaging in planning for conservation laryngeal surgery. AB - Refined laryngeal conservation surgeries have been devised to maintain the voice while eradicating even extensive glottic and hypopharyngeal tumors. The role of diagnostic imaging is to adequately define the extent of disease in conjunction with endoscopy. Critical areas to be evaluated are the tongue base, vallecula, pre-epiglottic space, arytenoid cartilages, postcricoid region, hypopharynx, thyroid cartilage, and subglottis. PMID- 8726919 TI - Head and neck reconstruction. AB - Reconstructive procedures of the head and neck aim to repair soft tissue and bony defects while restoring optimal function and cosmesis. Defects necessitating reconstruction may be the result of congenital anomalies, destructive disease processes, trauma, or surgery. Because primary wound closure is often impossible in these settings and healing by secondary intention provides unacceptable appearance and function, tissues from elsewhere in the body are usually mobilized to achieve the reconstructive goals. Reconstructions using tissue grafts and flaps have unique characteristics on postoperative imaging that differ from the original tissues being replaced, as well as the appearance of the donor size prior to transposition. As these reconstructive techniques become more sophisticated, it is crucial that physicians have an appreciation for current reconstructive methods. PMID- 8726920 TI - Diagnostic imaging aids to head and neck radiation oncology. AB - Tumors of the head and neck are primary sites for radiation treatment. In combination with surgery, the role of radiation is further expanded; however, localization of the primary tumor and its metastases, actual or potential, and the relationship to critical structures are quintessential to the application of radiation. In this process, the information made available by appropriate imaging studies as interpreted by the diagnostic radiologist is a requirement for cancer management. PMID- 8726921 TI - Individualizing medical treatment for tobacco dependence. PMID- 8726922 TI - Adenosine deaminase (ADA) isoenzymes ADA1 and ADA2: diagnostic and biological role. PMID- 8726923 TI - Diagnostic criteria for the sleep apnoea syndrome: time for consensus. PMID- 8726924 TI - The costs of asthma. AB - At present, asthma represents a substantial burden on health care resources in all countries so far studied. The costs of asthma are largely due to uncontrolled disease, and are likely to rise as its prevalence and severity increase. Costs could be significantly reduced if disease control is improved. A large proportion of the total cost of illness is derived from treating the consequences of poor asthma control-direct costs, such as emergency room use and hospitalizations. Indirect costs, which include time off work or school and early retirement, are incurred when the disease is not fully controlled and becomes severe enough to have an effect on daily life. In addition, quality of life assessments show that asthma has a significant socioeconomic impact, not only on the patients themselves, but on the whole family. Underuse of prescribed therapy, which includes poor compliance, significantly contributes towards the poor control of asthma. The consequences of poor compliance in asthma include increased morbidity and sometimes mortality, and increased health care expenditure. To improve asthma management, international guidelines have been introduced which recommend an increase in the use of prophylactic therapy. The resulting improvements in the control of asthma will reduce the number of hospitalizations associated with asthma, and may ultimately produce a shift within direct costs, with subsequent reductions in indirect costs. In addition, costs may be reduced by improving therapeutic interventions and through effective patient education programmes. This paper reviews current literature on the costs of asthma to assess how effectively money is spent and, by estimating the proportion of the cost attributable to uncontrolled disease, will identify where financial savings might be made. PMID- 8726925 TI - Importance of baseline cotinine plasma values in smoking cessation: results from a double-blind study with nicotine patch. AB - Nicotine replacement by transdermal patches is more effective than placebo in smoking cessation, but has a low success rate after one year (9-18%). We tested whether this was attributed to insufficient nicotine replacement. We conducted a randomized trial to investigate the effect on outcome of different doses of transdermal nicotine replacement after stratification according to baseline plasma cotinine values. Two hundred and ninety seven adult smokers were enrolled. Those with baseline cotinine < or = 250 ng.ml-1 (low cotinine) were randomly assigned to placebo (LC-P) or to 15 mg 16 h nicotine patches (LC-15), and those with baseline cotinine > 250 ng.mL-1 (high cotinine) were randomly assigned to 15 mg (HC-15) or 25 mg (HC-25) 16 h nicotine patches. Plasma nicotine and cotinine values, expired carbon monoxide and withdrawal symptoms were measured at scheduled intervals during treatment. Smokers in the LC-15 group had a significantly higher success rate than placebo (28 vs 9%). Smokers with high baseline cotinine had lower success rates, and a high dose of nicotine did not increase success rate (HC-25 9% vs HC-15 11%). Subjects in the HC-15 group had the lowest percentage of nicotine replacement and a higher prevalence of withdrawal symptoms than the HC-25 group. Replacement was similar in groups LC-15 and HC-25, but the success rate was significantly lower in HC-25 group, despite similar levels of withdrawal symptoms. We conclude that a higher success rate was obtained after one year in smokers with low baseline plasma cotinine values. Determination of plasma cotinine values may be, thus, helpful in identifying smokers who could benefit from transdermal nicotine replacement. PMID- 8726926 TI - Increased content of hydrogen peroxide in the expired breath of cigarette smokers. AB - Cigarette smoking causes an influx of mononuclear phagocytes and polymorphonuclear leucocytes into the lower airways. These cells have altered oxygen metabolism and release more H2O2 than phagocytes from nonsmokers. In this study, we intended to determine whether asymptomatic cigarette smokers exhale more H2O2 than healthy nonsmokers. The content of H2O2 in the expired condensate of 27 nonsmokers and 33 cigarette smokers was measured spectrofluorimetrically (homovanillic acid method). The mean H2O2 level in the expired breath condensate of all cigarette smokers was about fivefold higher than that found in the whole nonsmoker group (0.24 +/- 0.32 versus 0.05 +/- 0.11 nM). However, only 16 smokers (49%) and 6 nonsmokers (22%) had detectable levels of H2O2 in expired breath that reached values 0.49 +/- 0.28 and 0.23 +/- 0.10 nM, respectively. Although the cigarette smoking status was similar for both male and female smokers, females expired 2.5 fold less H2O2 than males (0.15 +/- 0.24 (n = 21) versus 0.38 +/- 0.39 (n = 12) nM. No correlation was found between expired H2O2 levels and cigarette smoking status expressed as the daily cigarette consumption, cumulative cigarette consumption and urinary cotinine concentration. It is suggested that in some smokers, expressed H2O2 can be a noninvasive marker of oxidant overload in the lower airways related to cigarette smoking. PMID- 8726927 TI - Prospective study of the value of transbronchial lung biopsy after lung transplantation. AB - Transbronchial lung biopsy (TBB) has become the gold standard for the diagnosis of acute rejection and cytomegalovirus (CMV) pneumonia in lung transplant recipients. The aim of this study was to assess the value of regular surveillance TBB in stable asymptomatic patients and to establish the role of TBB as a follow up procedure 1 month after a previous pathological biopsy result. We prospectively evaluated 76 TBBs performed in 17 lung transplant recipients. A definite pathological results was found in 14 of 15 TBBs performed for clinical indications: CMV pneumonia (5), acute rejection grade > or = A2 according to the criteria of the International Society for Heart and Lung Transplantation (ISHLT) (4), bronchiolitis obliterans (3), and desquamative interstitial pneumonitis (2). Fifteen of 45 surveillance TBBs performed in asymptomatic patients revealed significant abnormalities. Ten episodes of acute rejection ISHLT grade > or = A2 and three episodes of CMV pneumonia detected by TBB had direct therapeutic consequences. Nine of 16 follow-up TBBs performed 1 month after a pathological biopsy result again showed relevant pathological findings. With the exception of one severe haemorrhage, no life-threatening complications occurred. Our results suggest that transbronchial lung biopsies performed on a regular basis after lung transplantation are important for the detection of asymptomatic and/or persistent acute rejection or injection. In the long-term, this strategy might be the most effective tool in reducing the incidence of bronchiolitis obliterans, which is still the main obstacle for further improvement of long-term survival after lung transplantation. PMID- 8726928 TI - Pulmonary diffusion impairment following heart transplantation: a prospective study. AB - The aim of this prospective study was to confirm whether and when a fall in gas transfer occurs following heart transplantation (HT); and to examine the potential relationship between gas transfer and haemodynamic change, immuno suppression and cytomegalovirus (CMV) infection. The lung physiology of 34 heart transplant recipients (HTR) and 14 control patients undergoing coronary artery bypass grafting (CABG) were studied. The absolute and standardized residual values of forced expiratory volume in one second (FEV1), forced vital capacity (FVC), residual volume (RV), forced residual volume (FRC), total lung capacity (TLC), transfer factor of the lungs for carbon monoxide (TL,CO) and carbon monoxide transfer coefficient (KCO) were measured before and at 30, 60, 90, 120 and 150 days after HT. These data were compared to haemodynamic status, graft rejection, cyclosporin levels and episodes of CMV infection. Lung function was studied in a group of patients before and 4 weeks after CABG. There was a significant fall in mean KCO after HT (pre-HT = 1.29 and post-HT = 1.06 mmol.min 1.kPa.L-1) but not after CABG (pre-CABG = 1.49, post-CABG = 1.5 mmol.min-1.kPa.L 1. No relationship was observed between gas transfer and CMV. At the latest stage following HT (150 days) there was a positive correlation between TL,CO (absolute value and standardized residual) and mean cyclosporin level (r = 0.48 and r = 0.44, respectively) and also between the absolute KCO and actual (r = 0.56) and mean (r = 0.55) cyclosporin levels. Following HT, there is an early fall in gas transfer, which is independent of the effects of surgery and bypass, implicating early immunosuppression (e.g. antithymocyte globulin/cyclosporin). PMID- 8726929 TI - Relationships between radiological pattern and cell-mediated immune response in Mycoplasma pneumoniae pneumonia. AB - The aim of this study was to determine the relationship between the radiological pattern of Mycoplasma pneumoniae and the level of cell-mediated immunity of the host. Computed tomographic (CT) scans of the chest and the results of the purified protein derivative (PPD) test were studied during the acute stage of infection in 54 patients with M. pneumoniae pneumonia. The CT findings were used to divide the patients into two groups: one group had a predominance of nodular opacities with a centrilobular distribution (Group N; n = 29); and the other showed a predominance of an airspace consolidation (Group C; n = 25). Forty out of 54 subjects had negative tuberculin skin tests ( < 10 mm induration). The positive rate of PPD reaction was higher in Group N (13 out of 29) compared to Group C (1 out of 25) (p = 0.0005); whilst pleural effusion appeared more frequently in Group C (10 out of 25) than in Group N (3 out of 29) (p = 0.023). There was no significant difference between Groups N and C in white blood cell and lymphocyte counts, level of antibodies to M. pneumoniae in sera, and severity of the disease. These findings suggest that the characteristics of the host cell mediated immunity might influence the pattern of pulmonary lesions in M. pneumoniae infection. PMID- 8726931 TI - Relevance of asbestos bodies in sputum. AB - The presence of asbestos bodies (ABs) in sputum specimens of individuals with occupational asbestos exposure has been well-documented. The aim of this study was to determine their clinical relevance in comparison to the concentration of AB in bronchoalveolar lavage (BAL) and lung tissue. Subjects were included following a well-documented exposure of asbestos history (n = 93) or BAL fluid analysis positive for ABs (n = 42). The subjects with a well documented history of AB exposure were divided into three groups: heavy (Group 1, n = 29); moderate (Group 2, n = 31); or occasional exposure (Group 3, n = 33). BAL fluid was available from all subjects, and lung tissue from 21 subjects. To assess the variability, 10 sputum positive subjects collected subsequent sputum on days 2, 7, 14, 30 and 90. ABs were determined by light microscopy after membrane filtration of specimen digests. The mean sputum AB content was highest in Group 1 (2.4 +/- 5.5 AB.mL-1), lower in Group 2 (0.2 +/- 0.3 AB.mL-1) and lowest in Group 3 (0.1 +/- 0.1 AB.mL-1) suggesting a correlation with cumulative exposure. However, many negative sputum samples were noted, when BAL specimens were positive. The AB content of sputum and BAL specimens did not correlate. ABs were found in sputum of all subjects with a tissue content of > 1,000 AB-cm-3, but in none with contents of < 1,000 AB-cm-3. Substantial variability of ABs was found in the five sequentially collected sputa of 10 initially positive patients (coefficient of variation 28-93%), but only two false negatives were found in these 50 samples. Thus, sputum analysis for asbestos bodies is an insensitive method for assessing the lung asbestos burden, much less sensitive than bronchoalveolar lavage fluid analysis. However, a sputum sample positive for asbestos bodies is suggestive of a high lung asbestos burden. PMID- 8726930 TI - Integrin alpha E beta 7 expression on BAL CD4+, CD8+, and gamma delta T-cells in bleomycin-induced lung fibrosis in mouse. AB - CD4, CD8, and gamma delta T-cells located in the epithelium express the integrin alpha E beta 7 that binds to E-cadherin on the epithelium. Gamma delta T-cells mediate specific cellular immune functions and can recognize damaged cells directly. It was, therefore, of interest to analyse the presence of gamma delta T cells and the expression of alpha E beta 7 on gamma delta T-cells in the bleomycin (BLM) model of pulmonary fibrosis. Lung fibrosis was induced by a single intratracheal instillation of BLM (0.125 U.mouse-1), and bronchoalveolar lavage (BAL) T-cell subpopulations were examined at various time-points for the expression of the integrin alpha E beta 7 by flow cytometry. CD4+ T-cells accounted for about 40% of the lymphocytes, compared to about 10% of CD8+ T-cells and 10-14% gamma delta T-cells. Within the CD4+ T-cell population the proportion of alpha E beta 7+ cells decreased between Days 2 and 22 from 36 to 11%. The percentage of alpha E beta 7+ CD8+ T-cells increased at the same time from 4 to 68%. However, more than 80% of the gamma delta T-cells in BAL fluid expressed alpha E beta 7 at all time-points. The surface-expression of this integrin on gamma delta T-cells was 2-3 times higher than on CD4+ or CD8+ T-cells. This predominant expression of alpha E beta 7 on gamma delta T-cells suggests a role for these cells in the pathogenesis of bleomycin-induced lung fibrosis. PMID- 8726932 TI - Variations in the prevalence of respiratory symptoms, self-reported asthma attacks, and use of asthma medication in the European Community Respiratory Health Survey (ECRHS). AB - Asthma is a common chronic disorder which may be increasing in prevalence. However, little is known of its distribution and determinants. The European Community Respiratory Health Survey (ECRHS) is a multicentre survey of the prevalence, determinants and management of asthma. This paper presents a descriptive account of the variation in self-reported attacks of asthma and asthma symptoms across Europe, and in part fulfils the first aim of the study. A screening questionnaire, including seven questions relating to the 12 month prevalence of symptoms of asthma, was distributed to representative samples of 20 44 year old men and women in 48 centers, predominantly in Western Europe. The median response rate to the questionnaire was 75% but, after removing from the denominator those who were the wrong age, were known to have moved out of the area, or had died, it was 78% (range 54-100). The prevalence of all symptoms varied widely. Although these were generally lower in northern, central and southern Europe and higher in the British Isles, New Zealand, Australia and the United States, there were wide variations even within some countries. Centres with a high prevalence of self-reported attacks of asthma also reported high prevalences of nasal allergies and of waking at night with breathlessness. The use of asthma medication was more common where wheeze and asthma attacks were more frequent. In most centres in The Netherlands, Sweden, New Zealand and the United Kingdom over 80% of those with a diagnosis of asthma were currently using asthma medication. In Italy, France and Spain the rate was generally less than 70%. These data are the best evidence to date that geographical differences in asthma prevalence exist, are substantial and are not an artefact of the use of noncomparable methods. PMID- 8726933 TI - Anti-inflammatory effects of inhaled beclomethasone dipropionate in nonatopic asthmatics. AB - The effects of inhaled beclomethasone dipropionate (BDP) on asthma symptoms and infiltration of the bronchial mucosa by inflammatory cells were investigated in an open study of 10 patients with mild-to-moderate nonatopic bronchial asthma. Asthma scores were recorded in an asthma diary. Peak expiratory flow (PEF), PEF diurnal variation (PEF%), forced expiratory volume in one second (FEV1%), methacholine airway hypersensitivity (minimum dose of methacholine) (Dmin) were measured. Biopsy of the bronchial mucosa was performed before and after 8 weeks of treatment with BDP (400 micrograms.day-1). The following inflammatory cells were immunostained: eosinophils with anti-EG2; mast cells with AA1; neutrophils with NP57; T-lymphocytes with anti-CD3, CD4, and CD8; and activated T-lymphocytes with anti-CD25. There was a significant improvement in the asthma symptom score, PEF%, FEV1%, and Dmin after BDP therapy and the number of EG2-, AA1-, CD3-, CD4-, and CD25-positive cells decreased significantly. We conclude that inhaled beclomethasone dipropionate inhibited inflammatory cell infiltration of airway tissue and that associated with this there was an improvement of symptoms in this open study of inhaled beclomethasone dipropionate in a group of nonatopic asthmatic subjects. PMID- 8726934 TI - Priming of circulating human eosinophils following late response to allergen challenge. AB - The eosinophil is recognized as an important effector cell in asthma. To investigate the role of eosinophils in the aetiology of asthma, we have examined whether peripheral blood eosinophils are primed following whole-lung allergen challenge. Nineteen mild asthmatics, who each demonstrated a late asthmatic response (LAR), were studied. The priming of eosinophils following allergen challenge was assessed by measuring: 1) platelet-activating factor (PAF) and complement factor 5a (C5a) stimulated H2O2 release upon fibrinogen-coated tissue culture plates (n = 8); and 2) ex-vivo cell survival (n = 11). Subjects were venepunctured before and 24 h after allergen challenge and eosinophils were prepared by immunomagnetic separation. Basal H2O2 release was increased from eosinophils following allergen challenge. The response to PAF stimulation was significantly increased in terms of cell sensitivity (negative log of concentration producing half the maximal effect (pD2)) and responsiveness (maximum effect Emax). With C5a, although there was no change in sensitivity there was a significant increase in the maximal response. Ex-vivo eosinophil survival (at 3 days and cell half-life) was significantly prolonged following allergen challenge. In conclusion, we have demonstrated the priming of circulating eosinophils following allergen challenge. PMID- 8726935 TI - Airway structure and inflammatory cells in fatal attacks of asthma. AB - Fatal attacks of asthma usually occur against a background of chronic persistent symptoms, presumably due to chronic airway inflammation and changes in airway wall structure. Death from asthma is usually attributed to excessive airway narrowing due to a combination of muscle spasm and mucous plugging. To test the hypothesis that airway wall structure and/or the inflammatory cell profile are related to the duration of a fatal attack of asthma, inflammatory cell profiles and airway structure were examined in cases of fatal asthma and related to the duration of the fatal attack. In transverse sections of large and small airways from subjects dying from asthma, the numbers of eosinophils, neutrophils and lymphomononuclear cells were counted. The amount of smooth muscle shortening, the areas of airway wall, smooth muscle, mucous gland and cartilage were measured. Cell counts, airway dimensions and muscle shortening were compared in cases dying within 2 h of the fatal attack (short duration) and those dying more than 5 h after the onset of the fatal attack (long duration). In cases with fatal attacks of short duration, the numbers of neutrophils and the mucous gland area were increased and the numbers of eosinophils were reduced compared to cases with fatal attacks of long duration. Lymphocyte numbers, airway wall thickness, the areas of smooth muscle and cartilage and the amount of smooth muscle shortening were similar in the two groups. These findings suggest fatal attacks of asthma may be triggered by an inflammatory stimulus and suggest that increased production of mucous may contribute to sudden death in such cases. PMID- 8726936 TI - Mucociliary clearance during and after isocapnic hyperventilation with dry air in the presence of frusemide. AB - We have previously shown that mucociliary clearance (MCC) decreased during and increased after isocapnic hyperventilation (ISH) with dry air, both in asthmatic and healthy subjects. Inhaled frusemide, an inhibitor of the Na+/K+/2Cl- and NaCl co-transporters on the basolateral membrane of the epithelial cell, prevents the airway narrowing provoked by ISH with dry air. The co-transport system controls epithelial cell volume and chloride secretion and, thus, frusemide has the potential to modify the rate of recovery of periciliary fluid volume during and after ISH with dry air, and hence affect MCC. Frusemide also blocks mediator release from mast cells, which may also modify the increase in MCC after ISH. Eleven asthmatic and 11 healthy subjects inhaled frusemide (35.7 +/- 0.44 mg) or its vehicle, from a Fisoneb ultrasonic nebulizer 30 min before ISH with dry air, on two separate occasions. MCC was measured using 99mTc-sulphur colloid and a gamma camera. Frusemide, compared to its vehicle, did not affect MCC during or 45 min after ISH. However, in the presence of frusemide, the onset of the increase of MCC after ISH was significantly delayed for approximately 10 min in the whole right lung (p < 0.002) and central region (p < 0.01) in the asthmatic but not in the healthy subjects. These findings could be explained by frusemide delaying the recovery of the periciliary fluid volume after ISH with dry air and/or interfering with the stimulus that causes the increase in MCC in the asthmatic subjects after ISH. PMID- 8726937 TI - Inhalation of hypertonic saline aerosol enhances mucociliary clearance in asthmatic and healthy subjects. AB - Hyperosmolarity of the airway surface liquid (ASL) has been proposed as the stimulus for hyperpnoea-induced asthma. We found previously that mucociliary clearance (MCC) was increased after isocapnic hyperventilation (ISH) with dry air, and we proposed that the increase related to transient hyperosmolarity of the ASL. We investigated the effect of increasing the osmolarity of the ASL on MCC, by administering an aerosol of concentrated salt solution. MCC was measured using 99mTc-sulphur colloid, gamma camera and computer analysis in 12 asthmatic and 10 healthy subjects on three separate days, involving administration of each of the following: 1) ultrasonically nebulized 14.4% saline; 2) ultrasonically nebulized 0.9% saline; and 3) no aerosol intervention (control). The (mean +/- SD) volume of nebulized 14.4% saline was 2.2 +/- 1.2 mL for asthmatics and 3.2 +/ 0.7 mL for healthy subjects. This volume was delivered over a period of 5.4 +/- 1.3 and 6.4 +/- 0.7 min for asthmatic and healthy subjects, respectively. The airway response to 14.4% saline was assessed on a separate visit and the fall in forced expiratory volume in one second (FEV1) was 22 +/- 4% in the asthmatic and 3 +/- 2% in the healthy subjects. Compared to the MCC with the 0.9% saline and control, the hypertonic aerosol increased MCC in both groups. In asthmatic subjects, MCC of the whole right lung in 1 h was 68 +/- 10% with 14.4% saline vs 44 +/- 14% with 0.9% saline and 39 +/- 13% with control. In healthy subjects, MCC of the whole right lung in 1 h was 53 +/- 12% with 14.4% saline vs 41 +/- 15% with 0.9% saline and 36 +/- 13% with control. We conclude that an increase in osmolarity of the airway surface liquid increases mucociliary clearance both in asthmatic and healthy subjects. These findings are in keeping with our previous suggestion that the increase in mucociliary clearance after isotonic hyperventilation with dry air is due to a transient hyperosmolarity of the airway surface liquid. PMID- 8726938 TI - Immediate and delayed effects of nitrogen dioxide exposure at an ambient level on bronchial responsiveness to histamine in subjects with asthma. AB - The time-kinetics of NO2 induced effects on bronchial responsiveness are poorly known as most observations have been made shortly after exposure. The aim of this study was to measure nonspecific bronchial responsiveness, lung function and inflammatory markers at different times after NO2 exposure in asthmatics. Nineteen subjects with mild asthma were exposed to either purified air or 488 micrograms.m-3 (0.26 ppm) NO2 for 30 min during intermittent exercise. Airway responsiveness to histamine, specific airway resistance (sRaw) and thoracic gas volume (TGV) were measured 30 min, 5 h, 27 h and 7 days after exposure. Peripheral blood inflammatory mediators and the expression of an adhesion molecule, (Mac1) on granulocytes, were analysed 30 min and 27 h after exposure. Bronchial responsiveness to histamine was significantly increased 5 h after NO2 exposure when compared to air (median provocative dose of histamine required to cause 100% increase of sRaw ((PDsRaw,100%) 110 micrograms after NO2 exposure vs 203 micrograms on air). There was a tendency for an increase after 30 min, which was nonsignificant (median PDsRaw,100% 100 vs 153 micrograms). NO2 exposure did not affect sRaw, but TGV was significantly reduced after exposure. We found an increased expression of Mac-1 on granulocytes 30 min after NO2 exposure when compared to pre-exposure values. No effect was seen on tryptase, eosinophil cationic protein (ECP), or myeloperoxidase (MPO). These results suggest that exposure to an ambient level of NO2 causes a delayed effect on bronchial responsiveness in asthmatics. The increased expression of an adhesion molecule in peripheral blood may indicate a NO2-induced priming of human granulocytes. PMID- 8726939 TI - Role of tachykinins in bronchoconstriction induced by intravenous administration of bradykinin in guinea-pigs. AB - To elucidate the role of tachykinins in bronchoconstriction induced by intravenous administration of bradykinin (Bk), we studied the effects of FK224, a neurokinin-1 (NK1) and neurokinin-2 (NK2) receptor antagonist, on the bronchoconstriction induced by intravenous (i.v.) administration of Bk (5-100 micrograms.kg-1) in guinea-pigs. Total pulmonary resistance -(RL) was measured using a pressure-volume sensitive body plethysmograph in anaesthetized artificially ventilated guinea-pigs pretreated with atropine (1 mg.kg-1) and propranolol (1 mg.kg-1). In the control group, i.v. administration of Bk produced a dose-dependent increase in RL. In animals pretreated with FK224, bronchoconstriction induced by higher doses of Bk (10, 50 and 100 micrograms.kg 1) was significantly reduced, whilst the bronchoconstriction caused by lower doses of Bk (5 and 7.5 micrograms.kg-1) was not. Pretreatment with a combination of FK224 and indomethacin markedly inhibited the bronchoconstriction induced by each dose of Bk compared with the groups pretreated with FK224 alone. Although pretreatment with indomethacin alone significantly reduced RL at a high dose of Bk (50 micrograms.kg-1), the reduction was significantly lower than that produced by a combination of FK224 and indomethacin. These results suggest that intravenous administration of a high dose of bradykinin causes bronchoconstriction both by cyclo-oxygenase products and by release of tachykinins. PMID- 8726940 TI - Adenosine deaminase (ADA) isoenzyme analysis in pleural effusions: diagnostic role, and relevance to the origin of increased ADA in tuberculous pleurisy. AB - The rise in adenosine deaminase (ADA) activity in the pleural fluid of tuberculous pleurisy patients, though used for diagnosis, is of unknown origin. In this work, we determined ADA activity and the activities of 2'-deoxyadenosine deaminase and ADA-2 in 350 patients. We also considered whether the results throw light on the origin of high pleural fluid ADA in tuberculous pleurisy and estimated the diagnostic efficiency of 2'-deoxyadenosine deaminase, ADA-2 and total ADA activities with and without the inclusion of the 2'-deoxyadenosine deaminase/ADA activity ratio in a combined criterion. The 350 pleural effusions were classified by previously established criteria as transudates (60 males/18 females) or as tuberculous (49 males/27 females), neoplastic (50 males/39 females), parapneumonic (36 males/19 females), empyematous (11 males/3 females), or miscellaneous (25 males/13 females) exudates. Total ADA, ADA-2 and 2' deoxyadenosine deaminase activities were, respectively, 127.5 +/- 2.9, 103 +/- 29.5 and 42.8 +/- 14 U.L-1 in tuberculous exudates. With diagnostic thresholds of 47, 40 and 22 U.L-1 respectively, the sensitivities of ADA, ADA-2 and 2' deoxyadenosine deaminase for tuberculosis were 100, 100 and 95%; their specificities 91, 96 and 92%; and their efficiencies 93, 97 and 93%, respectively. One hundred and one effusions (all 76 tuberculous, 12 neoplastic, 4 parapneumonic and 9 empyematous exudates) had total ADA levels > 47 U.L-1; of these, 8 neoplastic, 1 parapneumonic and all the tuberculous exudates had a 2' deoxyadenosine deaminase/ADA activity ratio < 0.49. The criterion of simultaneously having ADA > 47 U.L-1, ADA-2 > 40 U.L-1 and a 2'-deoxyadenosine deaminase/ADA activity ratio < 0.49 was satisfied by all the tuberculous effusions but only eight others (all neoplastic) (sensitivity 100%, specificity 97%, efficiency 98%). We conclude that: 1) high total ADA activity in tuberculous pleural effusions is due mainly to an increase in ADA-2, and, therefore, originated from the only known source monocytes and macrophages; 2) ADA-2 was a more efficient diagnostic marker of tuberculous pleurisy than total ADA activity, although the difference was not statistically significant; and 3) among effusions with high total ADA the 2'-deoxyadenosine deaminase/ADA activity ratio differentiates tuberculous effusions from empyemas and parapneumonic effusions, but fails to discriminate well between tuberculous and neoplastic effusions. PMID- 8726941 TI - In vitro modulation of induced neutrophil activation by different surfactant preparations. AB - Endotracheal surfactant administration has gained an important role in the treatment of respiratory failure. Polymorphonuclear neutrophil granulocyte (PMN) activation mediated by chemoattractants, such as interleukin-8 (IL-8), neutrophil activating peptide-2 (NAP-2) and formylated bacterial oligopeptides, has been found to be involved in the pathophysiology of acute respiratory failure. We investigated potential modulating effects of commercial surfactant preparations (Exosurf, Alveofact, Curosurf and Survanta) on spontaneous and chemoattractant induced PMN function. Isolated cytochalasin B (CytB)-treated PMNs from healthy adults were incubated with increasing concentrations of surfactant. The response of the cells was measured in terms of elastase release from the lysosomes within 30 min. The PMNs showed no direct activation by any of the surfactants tested. However, when cells were stimulated with suboptimal dosages of chemokines, such as IL-8 (2 nM) or NAP-2 (100 nM), or formyl-methionyl-leucyl-phenylalanine (fMLP) (50 nM), and co-incubated with increasing concentrations of surfactant (0.05-8 mg.mL-1) the release of elastase was markedly modulated depending on the surfactant preparation used. Whilst Exosurf and Alveofact showed only modest effects on the elastase release induced by all three mediators, Curosurf and Survanta markedly inhibited the cellular response in a dose-dependent manner. At concentrations above 1 mg.mL-1, Curosurf and Survanta decreased the IL-8-, NAP-2- and fMLP-induced elastase release by 83, 67 and 90%, and by 82, 75 and 80%, respectively. In conclusion, exogenous surfactant may modulate the inflammatory response of the airways by affecting the chemoattractant-induced polymorphonuclear neutrophil activation. Surfactant preparations with inhibiting properties on neutrophil activation may participate in the prevention of neutrophil-induced lung damage. PMID- 8726942 TI - Lazaroid U-74389F attenuates phorbol ester-induced lung injury in rabbits. AB - We examined the effect of the antioxidant lazaroid U-74389F on acute lung injury induced in rabbits by phorbol myristate acetate (PMA). Thirty minutes after receiving either U-74389F (15 mg.kg-1 i.v.) or U-74389F vehicle, rabbits (n = 60) were given PMA (60 micrograms.kg-1 i.v.). PMA vehicle injected rabbits (n = 20) served as controls. Over a 5 h period after PMA or PMA vehicle injection, we measured arterial pH, arterial oxygen tension (Pa,O2), arterial carbon dioxide tension (Pa,CO2), and the plasma concentration of the neutrophil chemoattractant interleukin-8 (IL-8). At postmortem, lungs were inspected for macroscopic injury and examined histologically. Malondialdehyde levels were assayed in lung tissue as an index of lipid peroxidation. In bronchoalveolar lavage (BAL), total and differential cell counts, protein and IL-8 concentrations were measured. Compared to normal controls, rabbits challenged with PMA alone developed arterial acidosis, hypercapnia and hypoxaemia, accompanied by significant rise in plasma IL-8 concentration. U-74389F pretreated animals did not develop significant arterial blood gas abnormalities and had significantly lower IL-8 concentration in plasma. U-74389F did not prevent PMA-induced lipid peroxidation. However, macroscopic signs of lung injury and the degree of alveolar haemorrhage and protein extravasation were significantly less severe in pretreated rabbits than in those given PMA alone. In addition, U-74389F significantly reduced IL-8 concentration and neutrophil number in BAL. By histological assessment, 80% of lung neutrophils were localized in alveolar spaces of animals receiving PMA alone. Conversely, in U-74389F pretreated animals, 75% of neutrophils were distributed within extra-alveolar blood vessels and alveolar septa. We conclude that lazaroid U-74389F attenuates lung injury in rabbits given PMA by preventing neutrophil migration into pulmonary alveoli. This effect may, in part, be related to downregulation of IL-8 production. PMID- 8726943 TI - Effects of rhDNase on purulent airway secretions in chronic bronchitis. AB - Recombinant human deoxyribonuclease (rhDNase) has been demonstrated to reduce in vitro the viscosity and to improve the transport capacity of purulent respiratory mucus in cystic fibrosis. During episodes of exacerbation of chronic bronchitis, the patients generally expectorate purulent mucus. Purulence of mucus is associated with an increased deoxyriboneucleic acid (DNA) concentration. We analyzed in vitro the potential effect of rhDNase on chronic bronchitis mucus transport by the ciliary activity (frog palate model) and by simulated cough (cough machine model), as well as the effect on mucus viscosity (controlled stress rheometer) and surface properties (contact angle). Purulent sputa collected from patients with chronic bronchitis (n = 15) during an episode of exacerbation were incubated for 30 min at 37 degrees C with either rhDNase at two different concentrations (final concentration 2 or 4 micrograms.mL-1) or placebo. The median mucociliary transport rate was significantly improved by rhDNase from 0.68 with placebo to 0.79 and 0.83 with 2 and 4 micrograms.mL-1 of rhDNase, respectively. A significant improvement in mucus cough transport was also induced by rhDNase from 25.5 mm with placebo to 27.0 mm with either 2 or 4 micrograms.mL 1 rhDNase. These improvements in mucus transport capacity were associated with alterations in the physical properties of the mucus. The mucus median control viscosity (511.4 Pa.s) and median contact angle (0.85 rd) significantly decreased to 112.5 Pa.s and 0.74 rd, respectively, in the presence of 4 micrograms.mL-1 of rhDNase. These findings demonstrate that recombinant deoxyribonuclease may exert a beneficial effect on mucus clearance in vitro by altering the viscosity and surface properties of the purulent chronic bronchitic sputum samples. PMID- 8726944 TI - Tracheal distensibility in cystic fibrosis. AB - Size and distensibility of large airways have important implications for flow limitation and the efficacy of coughing. From radiological and functional data, some authors have suggested an increased size and distensibility of the trachea in cystic fibrosis (CF). Using computed tomography (CT) we compared size and distensibility of the trachea in 5 cystic fibrosis patients and five age- and height-matched healthy volunteers. Tracheal cross-sectional area was measured 25 mm below the cricoid cartilage. CT recordings were made at functional residual capacity, at 0 and +20 cmH2O mouth pressure. Inductive plethysmography was used to check that during these manoeuvres lung volume did not change and that the glottis remained open. Tracheal cross-sectional area and derived indices of tracheal distensibility were similar in the two groups. This study does not support the concept of an increased size and distensibility of the trachea in cystic fibrosis. PMID- 8726945 TI - Palatopharyngeus has respiratory activity and responds to negative pressure in sleep apnoeics. AB - The intrinsic tongue muscle, genioglossus, and soft palatal muscles, tensor palatini, levator palatini and palatoglossus, are known to exhibit phasic respiratory activity and to respond reflexly to negative pressure in man, which may be important in the maintenance of upper airway patency. We hypothesized that the palatopharyngeus muscle should also have respiratory activity and increased activity in response to negative upper airway pressure. We have, therefore, examined the palatopharyngeus and the antagonist muscle, levator palatini, in eight awake sleep apnoea patients, using bipolar electromyography during nose- or mouth-breathing in different postures, and with or without application of negative pressure. Overall, palatopharyngeus and levator palatini demonstrated phasic respiratory activity. Palatopharyngeus demonstrated increased activity in the supine compared to erect posture. Analysis by route of respiration showed palatopharyngeus to be more active in the supine posture when nose-breathing compared to mouth-breathing. Graded negative pressure application caused significant increases in palatopharyngeus and levator palatini activity when applied via the mouth. Nasal negative pressure application caused a nonsignificant trend to increasing palatopharyngeus activity with increasing negative pressure compared to a significant increase for levator palatini. We conclude that palatopharyngeus and levator palatini demonstrate respiratory activity and reflex activation in response to negative pressure. Palatopharyngeus may, therefore, have a role as an upper airway dilator. The differential response of palatopharyngeus to oral and nasal negative pressure application may be important in the pathogenesis of sleep apnoea. PMID- 8726946 TI - Expiratory flow limitation during mechanical ventilation detected by the forced oscillation method. AB - We have previously observed large phasic variations of respiratory mechanical impedance in chronic obstructive pulmonary disease (COPD) patients mechanically ventilated for acute respiratory failure, and postulated that they were due to expiratory flow limitation (EFL). The aim of this study was to test that assumption experimentally and to assess the value of impedance for automatic and noninvasive detection of EFL during mechanical ventilation. The study was performed: 1) in a mechanical analogue, including a flow-limiting element; and 2) in eight anaesthetized and paralysed rabbits, before and during histamine infusion. In both instances, EFL was obtained by lowering the expiratory pressure, using a computer-controlled ventilator; the absence of flow increase when expiratory pressure was further lowered was taken as evidence of EFL. Impedance was measured by applying 15 Hz oscillations at the airway opening. Its real (Re) and imaginary (Im) parts were measured separately during the inspiratory and the expiratory phases, and their differences were related to the mean inspiratory modulus. With the analogue, EFL was accompanied by large decreases both of Re and Im during the expiratory phase. In the rabbits, phasic variations of Re were variable in sign and were not significantly different with and without EFL. In contrast, EFL systematically and specifically decreased Im during the expiratory phase. A threshold of -50% provided a sensitivity of 96% and a specificity of 100% for detecting EFL. The observed phasic variations may be explained by airway wall shunt properties. The study suggests that a large decrease of the imaginary part of impedance during the expiratory phase is a sensitive and specific index of expiratory flow limitation during artificial ventilation. PMID- 8726947 TI - Pulmonary hypertension in the obstructive sleep apnoea syndrome: prevalence, causes and therapeutic consequences. AB - "Cor pulmonale" is a classic feature of the "Pickwickian syndrome". Earlier studies have reported a high prevalence of pulmonary hypertension (PH) in obstructive sleep apnoea (OSA) patients, but this has not been confirmed by recent studies with a more adequate methodology, including larger groups of patients. The first part of this review is devoted to the prevalence of PH in OSA; most recent studies agree on prevalence of 15-20%. The second (and major) part of the study deals with the causes and mechanisms of PH in OSA. Pulmonary hypertension is rarely observed in the absence of day-time hypoxaemia, and the severity of nocturnal events (apnoea index (AI), apnoea+ hypopnoea index (AHI) does not appear to be the determining factor of PH. Diurnal arterial blood gas disturbances and PH are most often explained by the presence of severe obesity (obesity-hypoventilation syndrome) and, principally, by association of OSA with chronic obstructive pulmonary disease (the so called "overlap syndrome"). Bronchial obstruction is generally of mild-to-moderate degree and may be asymptomatic. The final part of the review analyses the therapeutic consequences of the presence of PH in OSA patients. Pulmonary hypertension, which is generally mild-to-moderate, does not need a specific treatment. When nasal continuous positive airway pressure (CPAP) fails to correct sleep-related hypoxaemia, supplementary oxygen must be administered. In patients with marked daytime hypoxaemia (arterial oxygen tension (Pa,O2), < or = 7.3 kPa (55 mmHg) conventional O2 therapy (nocturnal + diurnal) is required. PMID- 8726948 TI - Noninvasive mechanical ventilation in acute respiratory failure. AB - Mechanical ventilation (MV) has been indicated in the treatment of acute respiratory failure (ARF) if conservative treatment fails. Invasive MV is associated to a variety of complications. The recent innovations of noninvasive methods of MV (NMV) avoid the complications of invasive MV, whilst ensuring a similar degree of efficacy. A review of the literature from 1989 to 1995 shows that use of NMV in ARF has been reported in several studies involving more than 400 patients most of them COPD. NMV was successful from 51 to 91%, the severity of ARF being widely different among the different studies. Most of the studies compared effectiveness of NMV with historical groups of patients treated with "conventional" medical therapy whilst controlled studies of NMV versus ET intubation are lacking. Type of mask, mode of ventilation, compliance to treatment, type of patient and severity of disease may influence the success rate. Success with NMV was associated with less severely abnormal baseline clinical and functional parameters and to less severe levels of acidosis assessed during an initial trial of NMV. Therefore, NMV may be useful in selected patients with ARF. Patients should have clinical and physiological evidence of ARF and should be sufficiently cooperative. It is commonly said that NMV should be avoided, and endotracheal (ET) intubation performed in patients with haemodynamic instability, uncontrolled arrhythmias, gastrointestinal bleeding, high risk for aspiration. With these limitations NMV in selected patients with ARF is well tolerated and may be useful in avoiding ET intubation in most cases of COPD and with a wide range of success rates in other disease. This in turn has several advantages in terms of avoiding complications of invasive MV, reducing the length of stay in ICU and probably the number of ICU readmissions. Side effects of NMV seem less severe than those induced by invasive MV. In conclusion in selected patients a trial of noninvasive mechanical ventilation, as an adjunct to medical therapy, should be instituted at an early stage of ARF episodes before severe acidosis ensures, to avoid ET intubation. PMID- 8726949 TI - Airway smooth muscle cell culture: application to studies of airway wall remodelling and phenotype plasticity in asthma. AB - Chronic persistent asthma is characterized by poorly reversible airway obstruction. Histopathological studies of airways removed postmortem from patients with severe asthma reveal marked inflammatory and architectural changes associated with airway wall thickening. Increased airway smooth muscle content, occurring as a result of hyperplastic and/or hypertrophic growth, is believed to be one of the principal contributors to airway wall thickening. Intense interest is building to discover the mechanisms responsible for these long-term structural changes. In vitro cell culture offers a powerful and exacting approach to cellular and molecular studies of the long-term regulation of airway smooth muscle function. This review discusses the methodologies for establishing and maintaining cell cultures of airway smooth muscle. It also describes the characteristics of these cells in culture and addresses the potential importance of phenotype plasticity and its possible relationship to altered smooth muscle function in vivo. Drawing on parallels from vascular studies, this review focuses, in particular, on the synthetic nature of the airway smooth muscle cell, emphasizing its potential to alter the composition of the extracellular matrix environment and orchestrate key events in the process of chronic airway remodelling. PMID- 8726950 TI - Long-, intermediate- and short-term growth studies in asthmatic children treated with inhaled glucocorticosteroids. AB - During recent years, new auxological techniques have been introduced for assessment of the risk of growth suppression in asthmatic children treated with exogeneous glucocorticosteroids. Assessment of lower leg growth rates with the knemometer has made short-term studies of growth processes under strictly controlled conditions possible. However, short-term lower leg growth rates cannot be used for estimations of intermediate-term height growth rates or long-term evaluations of final height. Consequently, the distinctions between the various types of growth studies in asthmatic children treated with inhaled glucocorticosteroids have become important and need to be discussed. The present paper presents a review of the long-, intermediate- and short-term growth studies available. The bulk of evidence from intermediate- and short-term evaluations indicates that growth rate is not affected when standard paediatric doses of inhaled glucocorticosteroids are used. However, further focus needs to be placed on differences between specific glucocorticosteroids, doses and delivery systems. Intermediate- and short-term growth data should be evaluated in the long-term perspective. PMID- 8726951 TI - Peak expiratory flow and the resistance of the mini-wright peak flow meter. AB - The purpose of this study was to examine whether the resistance of the peak flow meter influences its recordings. One hundred and twelve subjects, (healthy nonsmokers and smokers and subjects with lung diseases) performed three or more peak expiratory flow (PEF) manoeuvres through a Fleisch pneumotachograph with and without a mini-Wright peak flow meter added in random order as a resistance in series. The results were as follows. In comparison with a pneumotachograph alone, peak flow measured with an added mini-Wright meter had a smaller within-test variation, defined as the difference between the highest and second highest values of PEF in a series of blows. The mean (SE) variation was 14 (1.3) L.min-1 and 19 (1.5) L.min-1 with and without meter added, respectively. In comparison with the pneumotachograph alone, the addition of the mini-Wright meter caused PEF to be underread, especially at high flows. The difference (PEF with meter minus PEF without meter) = -0.064 (average PEF) -8 L.min-1; R2 = 0.13. The mean difference was -7.8 (1.1) %, and increased numerically for a given PEF, when maximal expiratory flow when 75% forced vital capacity remains to be exhaled (MEF75%FVC) decreased. The reproducibility criteria for repeated measurements of peak flow are more appropriately set at 30 L.min-1 than the commonly used 20 L.min-1, because a within-test variation of less than 30 L.min-1 was achieved in 76% of the subjects without PEF meter inserted and in 88% with meter inserted, with no difference between healthy untrained subjects and patients. The resistance of the peak expiratory flow meter causes less variation in recordings but reduces peak expiratory flow, especially at high values and when the peak is large as compared with the rest of the maximal expiratory flow-volume curve. PMID- 8726952 TI - Supplemental oxygen and nasal intermittent positive pressure ventilation. AB - When supplementary oxygen is necessary with nasal intermittent positive pressure ventilation (NIPPV), the optimal route by which it should be added to the ventilator circuit is unknown. We investigated the oxygen concentration received when oxygen was supplied at flow rates between 0 and 6 L.min-1 into the proximal ventilator tubing or the nasal mask whilst patients were ventilated with air. Eleven patients with stable chronic hypercapnic respiratory failure were studied. A calibration curve was produced for each by supplying different known oxygen concentrations through a Monnal D or DCC ventilator and measuring the arterial oxygen saturations achieved. Oxygen was then supplied into the ventilator tubing or nasal mask and arterial saturation again measured. The oxygen concentration received was estimated using the calibration curve. Tracheal oxygen concentration throughout the respiratory cycle was studied in one patient when oxygen was supplied by both routes. Peak inspired oxygen concentration occurred at end inspiration when oxygen was supplied into the ventilator tubing, but at mid inspiration when supplied into the nasal mask. However, there was no significant difference between the two routes in the inspired oxygen concentration achieved at all flow rates: 1 L.min-1 supplied approximately 31% oxygen; 2 L.min-1 37%; 3 L.min-1 40%; and 4 L.min-1 44%. Flow rates above 4 L.min-1 had little additional effect. In conclusion, oxygen supplementation during nasal intermittent positive pressure ventilation can be provided into the ventilator tubing or the nasal mask with equal efficiency. PMID- 8726953 TI - Cryptococcosis: an unusual cause of endobronchial obstruction. AB - We report the case of a 43 year old male patient, with normal immune function, who presented with right middle and lower lobe collapse. At bronchoscopy, a white lobulated lesion was seen, completely obstructing the origin of bronchus intermedius. Bronchial washings and biopsy of the lesion demonstrated cryptococcal organisms. The patient responded clinically and radiologically to amphotericin B and flucytosine; however, repeat bronchoscopy revealed only partial resolution of the endobronchial lesion. PMID- 8726954 TI - Mesenchymal chondrosarcoma of the pleura. AB - A 52 year old man developed an extraskeletal mesenchymal chondrosarcoma (ESMC) arising from the pleura. Clinically, the tumour mimicked a mesothelioma. Fine needle biopsy was consistent with the diagnosis of sarcomatoid mesothelioma. Histological examination of multiple tumour samples, supported by immunohistochemical characterization, made it possible to correctly diagnose extraskeletal mesenchymal chondrosarcoma. PMID- 8726955 TI - Persisting unilateral lung hypoperfusion after resection of a subcarinal bronchogenic cyst. AB - As a rare malformation with a wide variety of clinical modes of presentation, the bronchogenic cyst remains a diagnostic challenge. We report a case with a subcarinal bronchogenic cyst and stenosis of the left main bronchus presenting as "unilateral hyperlucent lung". Hypoplasia of the ipsilateral pulmonary vascular system persisted after surgical restitution of ventilation and is most likely due to an associated vascular malformation. Alternatively, anatomical fixation of pulmonary reflex vasoconstriction can be discussed. PMID- 8726956 TI - A patient with haemoptysis and a smaller right lung. PMID- 8726957 TI - Predictors of pulmonary hypertension in obstructive sleep apnoea syndrome. PMID- 8726958 TI - Chest physical examination. PMID- 8726959 TI - Characterization of a voltage-dependent L-type calcium channel from rabbit and turtle brain. AB - The binding of [3H]nitrendipine to membrane preparation from turtle and rabbit brain was studied. A single population of [3H]nitrendipine binding sites was detected in both species. [3H]nitrendipine bound with high affinity to brain membrane from both rabbit and turtle, revealing a significant population of binding sites (K(D) values of 0.55 +/- 0.05 nM and 0.56 +/- 0.04 nM and Bmax values of 122 +/- 11 and 275 +/- 18 fmol/mg of protein, respectively). Displacement studies showed a similar order of potency of various unlabeled ligands against [3H]nitrendipine both in rabbit or in turtle: nitrendipine > nifedipine >or= nicardipine >> verapamil >or= diltiazem. Our results show that a two fold increment of [3H]nitrendipine binding sites exists in the turtle brain respect to the rabbit. PMID- 8726960 TI - Influences of hypothyroidism on lipid composition and inositol lipid-linked receptors responsiveness and protein kinase C (PKC) activity in the cerebral cortex of Lewis rats. AB - The influence of hypothyroidism (HO) induced by treatment with propylthiouracil on lipid composition, receptor responsiveness of M1-muscarinic receptors (M1AChRs) and metabotropic glutamate receptors (mGluRs) as well as on protein kinase C (PKC) activity was investigated in the cerebral cortex of Lewis rats. HO did not influence the lipid composition. There was a significant 2-fold increase of efficacy and 6-fold decrease of potency of carbachol-induced inositol phosphate (IP) accumulation in HO, with respect to control rats. The efficacy of trans-(1S,3R)-1-amino-1,3-cyclopentanedicarboxylic acid (ACPD)-induced IP accumulation was also higher in HO (by 50%), without differences in EC50 values. The activities of soluble calcium-dependent and calcium-independent PKC were higher in HO than in control rats (both roughly 30%); membrane-associated PKCs were not modified. The data indicate that HO induces an increased responsiveness of M1AChRs and mGluRs and a rise in the soluble PKC activity that could be available and ready for translocation. PMID- 8726961 TI - Evidence for an in vivo and in vitro modulation of endogenous cortical GABA release by alpha-glycerylphosphorylcholine. AB - The effects of alpha-glycerylphosphorylcholine (alpha-GPC) on endogenous cortical GABA release were studied both in vivo and in vitro. In freely moving rats, equipped with epidural cups, alpha-GPC (30-300 mg/kg i.p.) increased GABA release. This effect was potentiated by atropine, both systematically administered (5 mg/kg i.p.) and locally applied (1.4 microM), but not by mecamylamine (4 mg/kg i.p.). The alpha-GPC-induced increase in GABA release was abolished in rats pretreated with the alpha 1 receptor antagonist prazosin (14 micrograms/kg i.p.). In cortical slices alpha-GPC (0.4 mM) increased the spontaneous GABA efflux. This effect was abolished by tetrodotoxin (0.5 microM) and prazosin (1 microM), but not by atropine (0.15 microM) or mecamylamine (2.5 microM). These results indicate that the facilitatory response by alpha-GPC on GABA release does not depend on a direct activation of either muscarinic or nicotinic receptors, but suggest the involvement of the noradrenergic system. PMID- 8726962 TI - Signal transduction mechanism responsible for changes in axoplasmic transport caused by neurotransmitters. AB - Transduction mechanism for modulation of axoplasmic transport by neurotransmitters was studied using cultured mouse superior cervical ganglion cells. The transported particles were analyzed with a computer-assisted video enhanced differential interference contrast microscope system. Acetylcholine depressed and adrenaline increased axoplasmic transport. GTP-binding proteins linked with both receptors activate or inactivate adenylyl cyclase, thereby altering the intracellular concentration of cyclic AMP. The cyclic AMP activates protein kinase A, which phosphorylates certain enzymes and the enzymes in turn phosphorylate motor proteins. An inhibitor protein kinase A, KT5720, decreases the number of the transported particles. In a stable state the cyclic AMP level stays at a normal level. Treatment with neurotransmitters causes a change in this level, which changes the activity of protein kinase A and thus decreases or enhances the phosphorylation of motor proteins. These changes are involved in the modulation of axoplasmic transport. PMID- 8726963 TI - Enhancement of fluoxetine-dependent increase of extracellular serotonin (5-HT) levels by (-)-pindolol, an antagonist at 5-HT1A receptors. AB - The somatodendritic 5-HT1A autoreceptor is known to regulate activity of 5-HT neurons and consequently 5-HT release. Administration of a selective 5-HT uptake inhibitor, fluoxetine (10 mg/kg, i.p.) increased extracellular 5-HT levels in rat hypothalamus up to 260 percent of basal levels. (-)-Pindolol, and antagonist at the somatodendritic 5-HT1A autoreceptor, dose-dependently (1, 3 and 5 mg/kg, s.c.) potentiated the fluoxetine dependent increase up to 458 percent of basal 5 HT levels for approximately 1.5 hours. Continuous infusion of ( +/- )-pindolol at 30 mg/kg/h s.c. enhanced the fluoxetine dependent elevation of extracellular 5-HT concentrations in hypothalamus up to 464 percent of basal levels and lasted for 3 hours. Thus, the combination of 5-HT uptake inhibition with antagonism at the somatodendritic 5-HT1A autoreceptor can enhance 5-HT release to levels beyond those achieved with uptake inhibition alone. The present findings are consistent with the hypothesis that blockade of somatodendritic 5-HT1A autoreceptors removes the inhibitory effect exerted by the elevated 5-HT levels resulting from uptake inhibition. PMID- 8726964 TI - Evaluation of monoaminergic neurotransmitters in the acute focal ischemic human brain model by intracerebral in vivo microdialysis. AB - The release of neurotransmitters principally glutamate during cerebral ischemia has been extensively studied. It is well recognized that ischemia induced release of glutamate plays a key role in "excitotoxic" neuronal death. The role of monoaminergic neurotransmitters is however unclear. The purpose of this study was to evaluate the extracellular norepinephrine, 3,4-dihydroxyphenylacetic acid (DOPAC), 5-hydroxyindoleacetic acid (5-HIAA) and serotonin (5-HT) under varied degrees of ischemia in the acute focal ischemic model of the human brain by in vivo microdialysis. The ischemic response of these amines was correlated with the glutamate levels. Our study concludes that these amines and metabolites can be detected in the human "stroke" model. No marked fluctuations were noted in the levels of norepinephrine and DOPAC. However, significant changes to partial and total ischemia were noted in the extracellular levels of 5-HIAA and 5-HT. These compounds showed a dramatic increase with the onset of ischemia with higher detectable levels in the partial ischemic state in comparison to the total ischemic dialysate levels. The exact role played by the differential increase in the levels of 5-HT to the other catecholamines in the pathogenesis of ischemic neuronal damage remains unclear and warrants further study. PMID- 8726965 TI - Effects of ammonia on L-glutamate uptake in cultured astrocytes. AB - The effect of ammonia on L-glutamate (L-GLU) uptake was examined in cultured astrocytes. Acute ammonia treatment (5-10 mM) enhanced L-[3H]GLU uptake by 20-42% by increasing the Vmax; this persisted for 2 days and than started to decline. Ammonia, however, did not affect the uptake of D-[3H]aspartate (D-ASP), a non metabolizable analog of L-GLU, that uses the same transport carrier as L-GLU. Also, L-GLU uptake was not affected during the first 2 min of the assay. Thus, ammonia did not have an acute effect of L-GLU transport (translocation); rather, ammonia enhanced the accumulation or "trapping" of L-GLU or its by-products. Chronic ammonia treatment, on the other hand, inhibited L-GLU transport in astrocytes by approximately 30-45% and this was due to a decrease in Vmax, suggesting that the number of L-GLU transporters was decreased. This inhibitory effect was observed after 1 day of treatment and persisted for at least 7 days. The inhibition of L-GLU transport was partially reversible following removal of ammonia. The effects of ammonia on L-GLU transport and uptake may explain the abnormal L-GLU neurotransmission observed in hyperammonemia/hepatic encephalopathy, and the brain swelling associated with fulminant hepatic failure. PMID- 8726966 TI - Serotonergic activity of HP 184: does spontaneous release have a role? AB - Examination of HP 184, [N-n-propyl)-N-(3-fluoro-4-pyridinyl) -1H-3-methylindodel 1-amine hydrochloride], in a variety of tests for serotonergic activity revealed some unique properties of this compound. We report here that 100 microM HP 184 enhanced spontaneous release of [3H]serotonin (5-HT) from rat hippocampal slices. This release was independent of the uptake carrier. In vivo assays confirmed that HP 184 (20 mg/kg, i.p.) lacked significant interactions at the norepinephrine (NE) or 5-HT uptake carrier itself. Notably, HP 184 (15 mg/kg, i.p.) reduced drinking behavior in schedule-induced polydipsic (SIP) rats. We previously reported that some selective 5-HT reuptake inhibitors decrease SIP 30-40% after a 14-21 day treatment. In the current study, HP 184 decreased SIP beginning with the first treatment, and this reduction (30%) was maintained for 28 days. We further investigated HP 184 and serotonin metabolite levels. One hour after i.p. administration of 30 mg/kg HP 184, the ratio of whole brain 5-hydroxyindolacetic acid (5-HIAA) to 5-HT was increased, suggesting serotonergic activation. Under these conditions, the brain:plasma ratio of HP 184 was approximately 2:1, with brain concentrations of 1.6 micrograms/gram. We speculate that the spontaneous release effects of HP 184 may be responsible for the behavioral effects observed. PMID- 8726967 TI - Synaptic degeneration and remodelling after fast kindling of the olfactory bulb. AB - Kindling of the olfactory bulb using a novel fast protocol (within 24 h) was studied in rats. In target brain regions, the effects of kindling were measured on the concentration of glial fibrillary acidic protein (GFAP) by dot-blot and on the concentrations of neural cell adhesion molecule (NCAM) and the 25 kDa synaptosomal associated protein of the D3 immunoprecipitate (D3(SNAP-25)) by crossed immunoelectrophoresis. Bilateral increases in the levels of GFAP, indicating activation of astrocytes, were detected in primary olfactory cortical projection areas, including the piriform cortex, and also in the basolateral amygdala and dentate gyrus, suggesting that these regions may be functionally altered during the kindling process. In the piriform cortex and dentate gyrus increased NCAM/D3(SNAP-25) ratios found ipsilaterally at seven days after kindling probably reflect an elevated rate of synaptic remodelling. At this time, however, an overall pattern of ipsilateral decreases in the synaptic marker proteins NCAM and D3(SNAP-25) indicated that this remodelling occurred on a background of synaptic degeneration. These results confirm previous studies showing that kindling is associated with synaptic remodelling and neuronal degeneration in the hippocampal formation and extends the area of plasticity to include the piriform cortex which is believed to be central to the kindling process. PMID- 8726968 TI - Effects of brain ischemia on intermediate filaments of rat hippocampus. AB - Neurofilaments subunits (NF-H, NF-M, NF-L) and glial fibrillary acidic protein (GFAP) were investigated in the hippocampus of rats after distinct periods of reperfusion (1 to 15 days) following 20 min of transient global forebrain ischemia in the rat. In vitro [14Ca]leucine incorporation was not altered until 48 h after the ischemic insult, however concentration of intermediate filament subunits significantly decreased in this period. Three days after the insult, leucine incorporation significantly increased while the concentration NF-H, NF-M, and NF-L were still diminished after 15 days of reperfusion. In vitro incorporation of 32P into NF-M and NF-L suffered immediately after ischemia, but returned to control values after two days of reperfusion. GFAP levels decreased immediately after ischemia but quickly recovered and significantly peaked from 7 to 10 days after the insult. These results suggest that transient ischemia followed by reperfusion causes proteolysis of intermediate filaments in the hippocampus, and the proteolysis could be facilitated by diminished phosphorylation levels of NF-M and NF-L. PMID- 8726969 TI - Effectors of D-[3H]aspartate release from rat cerebellum. AB - The effect of aminooxyacetic acid (AOAA), NH4+, phenylsuccinate (Phs), ketone bodies (KB) and glutamine (Gln), that might interfere with the biosynthesis of neurotransmitter glutamate on the K(+)-evoked Ca(2+)-dependent release of D [3H]aspartate from rat cerebellar slices was studied. Therefore slices were preincubated in a Krebs-Ringer-bicarbonate-glucose (KR) buffer, loaded with D [3H]aspartate and superfused in the presence of Ca2+ or when Ca2+ was replaced by Mg2+ or in some cases by EGTA. AOAA, NH4+ and Phs increase the K(+)-evoked Ca(2+) dependent release of radioactivity by 30%, 68% and 188% compared to the control respectively indicating that these agents are inhibitors of the K(+)-evoked Ca(2+)-dependent release of glutamate. KB and Gln had no effect on the Ca(2+) dependent release of radioactivity. AOAA, NH4+, Phs and KB but not Gln increase the total release of radioactivity by 43%, 69%, 139%, and 37% respectively. AOAA, NH4+ and KB but not Phs or Gln increase the Ca(2+)-independent release (Mg2+ replacing Ca2+) of radioactivity by 71%, 71% and 108% respectively. The present results indicate that in the cerebellum: 1) Neurotransmitter glutamate is mostly synthesized through the phosphate activated glutaminase (PAG) reaction 2) It is further supported that glutamate released in Ca(2+)-dependent manner before entering its pool in the cytosol has to move into the mitochondrial matrix. PMID- 8726970 TI - Differential responsiveness of late passage C-6 glial cells and advanced passages of astrocytes derived from aged mouse cerebral hemispheres to cytokines and growth factors: glutamine synthetase activity. AB - In this study, we were interested to compare the responsiveness to growth factors, NGF, b-FGF and EGF and cytokines, IL1 beta, and TNF-alpha, in late passages (74-79) C6 glial cells committed astrocytes and astrocytes of advanced passages (26-28) in cultures derived from aged mouse cerebral hemispheres (MACH). Cultures were grown in either DMEM or chemically defined medium (CDM/TIPS) in order to test the effects of growth factors or cytokines. The activity of glutamine synthetase (GS), a marker for astrocytes, was used as a test parameter. We found that treatment with growth factors increased GS activity in both glial cell culture systems with the exception of EGF in C-6 glial cells. Treatment with cytokines markedly decreased GS activity in the late passage C6 glial cells whereas only TNF-alpha had a similar effect on MACH astrocytes. In view of the generally opposite effects of growth factors and cytokines on GS activity, we speculate that these molecules which are also endogenously present in glial cells may play a role in the maintenance of cellular homeostasis. PMID- 8726971 TI - Metabolism of N-acetyl-L-aspartate in rat brain. AB - The abundance and developmental regulation of N-acetylaspartate (NAA) in brain suggest that it plays an important role in brain metabolism. Previous studies demonstrated that NAA transports acetate from the mitochondrion to the cytoplasm where it is utilized for lipid synthesis, however, the metabolic fate of NAA derived aspartate is not established. To investigate NAA metabolism, rats were injected intracranially with N-([2H3]acetyl)-L-[15N]aspartate ([2H3,15N]NAA) and whole brain metabolites were analyzed using gas chromatography and mass spectrometry techniques (GC/MS). The rapid decline of [2H3,15N]NAA was associated with a rapid appearance of [15N]glutamate, indicating rapid transamination of the [15N]aspartate that was derived from the enzymatic hydrolysis of [2H3,15N]NAA. Inability to detect [15N]NAA in brain extracts in several experiments indicates that the 15N moiety is not reutilized for NAA synthesis and suggests one metabolic role of NAA may be the transport of amino nitrogen from the mitochondrion to the cytoplasm. PMID- 8726974 TI - Managed care: opportunities and challenges for occupational therapy in the emerging systems of the 21st century. AB - Overall, occupational therapy seems reasonably well situated to prosper in a cost conscious environment of the future. But the profession's standing in the next century will depend on its ability to build on its strengths and assist in cost containment by avoiding unnecessary or duplicative services and by fostering versatility through the education of generalists. Above all, the profession must recognize that patients will measure quality in terms of results and satisfaction related to the impact of their health on their everyday lives. PMID- 8726975 TI - Managed care fundamentals: implications for health care organizations and health care professionals. AB - Managed care is changing our health care delivery system as radically as the computer chip has changed telecommunications. Health care professionals and organizations that do not understand managed care's implications will not be prepared for the future. For example, one implication of managed care is payment capitation, which is the transfer of financial risk from the insurer to the provider. As a result, health care providers, including occupational therapy professionals, need to be better managers of scarce resources by recognizing the cost implications among various alternative procedures while still delivering quality care. Under managed care with capitation, occupational therapists will need to learn to provide services within the parameters of a fixed budget, requiring reengineering of the therapies and processes of care and a considerable reduction in the procedures and modalities for any given treatment or therapy. As a result, patients will be required to do more for themselves, and occupational therapists will have to become better patient educators and motivators. Additionally, managed care will require changes in professional curriculums, emphasis through continuing education, and assimilation of better cost information to practitioners to facilitate decision making. Implications of managed care other than payment capitation are assigning to enrollees a gatekeeper who is responsible for limiting access to costly specialty services, practicing utilization review to audit usage patterns and provide constructive recommendations to reduce costs and improve service quality, and forming networks and associations among medical providers for developing economies of scale and providing an integrated continuum of health care services to enrollees. PMID- 8726972 TI - Characterization and transplantation of two neuronal cell lines with dopaminergic properties. AB - Immortalized rat mesencephalic cells (1RB3AN27) produced dopamine (DA) at a level that was higher than produced by undifferentiated or differentiated murine neuroblastoma cells (NBP2) in culture. Treatment of 1RB3AN27 and NBP2 cells with a cAMP stimulating agent increased tyrosine hydroxylase (TH) activity and the intensity of immunostaining for the DA transporter protein (DAT). 1RB3AN27 cells were labelled with primary antibodies to neuron specific enolase (NSE) and nestin and exhibited very little or no labeling with anti-glial fibrillary acidic protein (GFAP). 1RB3AN27 cells exhibited beta- and alpha-adrenoreceptors, and prostaglandin E1 receptors, all of which were linked to adenylate cyclase (AC). Dopamine receptor (D1) and cholinergic muscarinic receptors linked to AC were not detectable. The levels of PKC alpha and PKC beta isoforms were higher than those of PKC gamma and PKC delta in 1RB3AN27 cells. The 1RB3AN27 cells were more effective in reducing the rate of methamphetamine-induced turning in rats with unilateral 6-OHDA lesion of the nigrostriatal system than differentiated NBP2 cells. The grafted 1RB3AN27 were viable as determined by DiI labelling, but they did not divide and did not produce T-antigen protein; however, when these grafted cells were cultured in vitro, they resumed production of T-antigen and proliferated after the primary glia cells and neurons of host brain died due to maturation and subsequent degeneration. Examination of H&E stained sections of the grafted sites revealed no evidence of infiltration of inflammatory cells in the grafted area suggesting that these cells were not immunogenic. They also did not form tumors. PMID- 8726973 TI - Inhibition of neuronal cyclin-dependent kinase-5 by staurosporine and purine analogs is independent of activation by Munc-18. AB - Neuronal cdk5 can phosphorylate certain lys-ser-pro (KSP) motifs of neurofilaments and tau protein in the nervous system. We have immunoprecipitated the cdk5 from rat brain using a polyclonal antibody raised against the C-terminus of cdk5. The immunoprecipitate has phosphorylated a KSPXK peptide analog of NF-H, as well as histone H1 and a bacterially expressed rat NF-H protein. The kinase activity was inhibited by staurosporine, isopentanyladenine and olomoucine in a dose dependent manner. Kinetic studies indicated Ki values of 39 nM, 38 microM and 8 microM, respectively for staurosporine, isopentanyladenine and olomoucine. The inhibition by staurosporine was non-competitive with respect to phosphoryl acceptor acceptor substrates. Western blot analysis of the immunoprecipitate showed both cdk5 and p67 (Munc-18), a putative regulator molecule of the kinase. Addition of p67 fusion protein enhanced the kinase activity of the immunoprecipitate by 60% above the basal activity. P67 elevated Ki values for both staurosporine and olomoucine. The degree of inhibition at high concentrations of these inhibitors was unaltered by exogenous p67 indicating a lack of competitive interactions with p67. The high affinity of staurosporine for cdk5 suggests that cdk5 may be one of the targets for the neurotropic effect of staurosporine. PMID- 8726976 TI - Development of critical paths for post-acute brain injury rehabilitation: lessons learned. AB - One important tool for case management is critical path analysis. This article explains four critical pathways developed by an interdisciplinary team for a post acute brain injury rehabilitation program. The heterogeneity of the brain injury population mandates the need for systematic coordination of direct care services. Yet, variations in the neurobehavioral consequences of brain injury necessitate differing goals and treatment tracks for individual clients. The critical pathways in this setting define and describe the procedures and services to be rendered from admission to discharge to achieve optimal goals for four treatment program tracks: Return to Work, Return to School, Functional Independence, and Neurorehabilitation. The tracks reflect a hierarchy of expectations for information processing and functional performance. Critical pathways provide a tool for enhancing communication among service providers and external case managers and for determining the extent to which a client's course of treatment compares with a clinical standard considered to be ideal. This article compares the four critical pathways, provides representative case samples, and discusses lessons learned in the development and implementation process. PMID- 8726977 TI - Managed mental health care: reflections in a time of turmoil. AB - This article reviews the history, growth, and evolution of managed care in mental health and substance abuse treatment. Specific issues described are stigma, the important social dimensions and chronicity of some types of mental illness and chemical dependency, and reliance on the public sector for care. Opportunities and challenges for occupational therapists in the rapidly changing mental health system are discussed, including the use of interdisciplinary teams, the importance of measuring functional outcomes of interventions, the need to develop clinical guidelines, the importance of the community setting and a continuum of services, ethical dilemmas, and the importance of assertive occupational therapy advocacy and involvement in health care reform. PMID- 8726978 TI - Outcomes research: the history, debate, and implications for the field of occupational therapy. AB - The field of occupational therapy, as with all facets of health care, has been profoundly affected by the changing climate of health care delivery. The combination of cost-effectiveness and quality of care has become the benchmark for and consequent drive behind the rise of managed health care delivery systems. The spawning of outcomes research is in direct response to the need for comparative databases to provide results of effectiveness in health care treatment protocols, evaluations of health-related quality of life, and cost containment measures. Outcomes management is the application of outcomes research data by all levels of health care providers. The challenges facing occupational therapists include proving our value in an economic trend of downsizing, competing within the medical profession, developing and affiliating with new payer sources, and reengineering our careers to meet the needs of the new, nontraditional health care marketplace. PMID- 8726979 TI - Managing the occupational environment of managed care. AB - The emergence of managed care has resulted in a practice environment buffeted by rapid and sweeping changes. Shifts in philosophy and approach to payment for health care affect the nature of the service and the method of its delivery. The occupational adaptation frame of reference is used to illustrate the challenges imposed by the physical, social, and cultural components of the changing occupational environment. Perceptions, observations, and concerns of middle level occupational therapy, physical therapy, social work, and nursing managers illustrate the impact of this new environment. Therapists are challenged to understand their own changing work setting and to apply the same effort to mastering its challenges as they would to examining the context of their patient's occupational performance and to designing and implementing an appropriate clinical intervention. PMID- 8726980 TI - Interest among occupational therapy managers in measuring workload for case costing. AB - OBJECTIVES: Interest in costing health care delivery on an individual case basis has increased in recent years as concern with overall health care costs has heightened. Costing exercises have been largely oriented around medical classification systems. Measures to incorporate the contributions of allied health activities such as occupational therapy are relatively recent. The objective of this study was to examine the attitudes and opinions of senior occupational therapy managers toward workload measurement on the basis of case mix. METHOD: A survey was sent to all 198 senior occupational therapy managers in accredited Canadian facilities, which was completed by 182 respondents for a response rate of 92%. The questionnaire asked about workload measurement system(s) currently used, satisfaction with the system(s), and needs and expectations of workload systems in general. RESULTS: The majority of respondents were using a time recording workload measurement system and expressed relatively low levels of satisfaction with it. Current systems were unable to provide costing data, which respondents ranked as very important for themselves as managers. The majority of respondents believed that it would be useful or very useful to be able to cost occupational therapy services by a diagnostic grouping system and to establish standard protocols per diagnosis, standard times per procedure per institution, and standard times per procedure for the profession. CONCLUSION: There is support for developing or expanding current methods of measuring workload. Senior occupational therapy managers would like to be able to predict their workload prospectively, a step that will accommodate the move toward case costing and program management. PMID- 8726981 TI - Roles and functions of a case manager. PMID- 8726982 TI - Now that we have managed care, shall we inspire it? PMID- 8726983 TI - AOTA and managed care. PMID- 8726984 TI - Profit-managed health care. PMID- 8726985 TI - The resurgence of TB and its implications for radiology. AB - Tuberculosis is re-emerging as a major health threat. Between 1985 and 1990, the number of TB cases reported in the United States increased by more than 15%. In addition, multidrug-resistant strains of Mycobacterium tuberculosis have begun to proliferate. Multidrug-resistant tuberculosis is extremely difficult to treat and is associated with high mortality rates. Tuberculosis poses tremendous risk to certain populations, including HIV-positive or otherwise immunocompromised patients, the homeless and individuals in correctional facilities or nursing homes. Also, tuberculosis is the sixth most common occupationally acquired infection among laboratory workers. This article traces the etiology of tuberculosis and outlines diagnostic techniques and treatment modalities. It also discusses the role of radiologic technologists in screening patients with suspected tuberculosis. PMID- 8726986 TI - Effect of quantum mottle on radiographic image quality. AB - Any critique of radiographic image quality must include an evaluation of quantum mottle, a fundamental limitation of the imaging process. Using an illustrative analogy, this article explains the concept of quantum mottle and defines the quantum sink in an imaging chain. The article also describes the interdependence between quantum mottle and other system parameters, including spatial resolution, contrast resolution and total system speed. PMID- 8726988 TI - Acuity scheduling systems: a case study in radiology. AB - To improve efficiency, the special procedures division of a radiology department at a Florida hospital chose to implement an acuity system designed specifically to ease patient scheduling problems. The matrix-based acuity system minimized overtime hours for radiologic technologists, nurses and physicians while maximizing patient throughput and staff utilization. The acuity system was tested using General Purpose Simulation System (GPSS). Test results were positive, and the hospital plans to implement the system for patient scheduling. PMID- 8726987 TI - Responsible dissemination of scholarly work in radiology. AB - Scholarly research is critical not only to the technical advancement of the radiologic sciences, but also to the recognition of radiologic technology as a profession. As radiologic science professionals become increasingly involved in the production and publication of research, they need to understand the ethical implications of such research. This article describes the peer-review process, discusses problems of authorship, examines questionable research practices and presents a set of guidelines for the responsible dissemination of research. PMID- 8726989 TI - Re-engineering via patient-centered care. PMID- 8726990 TI - Standards are adopted by JRCERT. PMID- 8726991 TI - Using critical thinking to teach empathy. PMID- 8726992 TI - Applications of proton beam therapy. PMID- 8726993 TI - Localization of natriuretic peptide-activated guanylate cyclase mRNAs in the rat brain. AB - Physiological actions of atrial natriuretic peptide (ANP) and C-type natriuretic peptide (CNP) are elaborated by membrane-bound natriuretic peptide receptors (NPRs). These receptors possess intracellular guanylate cyclase domains that mobilize cyclic guanosine monophosphate upon binding of peptide. Two distinct NPR subtypes have been described in brain: the NPR-A selectively binds ANP, whereas NPR-B exhibits high affinity for CNP. To define further the potential domains of ANP and CNP action in brain, the present study used in situ hybridization histochemistry to map NPR-A and NPR-B mRNA-expressing cell populations. Significant levels of neuronal NPR-A mRNA expression were observed only in the mitral cell layer of the olfactory bulb, medial habenula, subfornical organ, and area postrema. Expression of NPR-A mRNA was observed in forebrain white matter tracts, suggesting synthesis in glial cells. In contrast, NPR-B mRNA was widely expressed throughout the neuraxis. In the telencephalon, signal was abundant throughout limbic cortex and neocortex, olfactory bulb, hippocampus, and amygdala. Intense NPR-B mRNA hybridization was observed in preoptic-hypothalamic neuroendocrine circuits and in motor nuclei of cranial nerves. Intermediate expression of NPR-B mRNA was observed in brainstem nuclei controlling autonomic function. Labeling for NPR-B but not NPR-A mRNA was observed in pituicytes in the neural lobe of the pituitary and in scattered cells of the anterior pituitary. These results suggest that CNP is the primary biologically active natriuretic peptide in brain. In contrast with NPR-B, NPR-A appears to be expressed largely in restricted cell populations containing high levels of ANP and in circumventricular organs. These data implicate the NPR-A in autoregulation of ANP neurons and central registration of cardiac ANP release. PMID- 8726994 TI - Calretinin-immunoreactive cells and fibers in the human amygdaloid complex. AB - Calretinin is a calcium-binding protein that colocalizes with GABA in the cerebral cortex and hippocampus of the rat and the monkey. In the present study, we investigated the distribution of calretinin-immunoreactive cells and fibers in the human amygdaloid complex. A conspicuous feature was the high density of calretinin neurons in the human amygdala. The highest densities of the calretinin immunoreactive neurons were observed in the anterior cortical nucleus, accessory basal nucleus, amygdalohippocampal area, and in the nucleus of the lateral olfactory tract. The paralaminar nucleus, central nucleus, medial nucleus, and the periamygdaloid cortex contained the lowest densities of calretinin neurons. In most of the amygdaloid areas, the calretinin cells had the appearance of aspiny or sparsely spiny local circuit neurons. However, in the amygdalohippocampal area, we found also densely spined dendrites. The cortical areas and the central nucleus were characterized by intense neuropil labeling, while the deep nuclei contained a high density of calretinin-immunoreactive fibers and terminals. Calretinin immunoreactivity was also found in the intra amygdaloid fiber bundles, stria terminalis, and in the ventral amygdalofugal pathway. This suggests that in addition to the local circuit neurons, calretinin immunoreactivity is also located in neurons that connect the amygdaloid complex with the other brain areas. The distribution and morphological characteristics of calretinin-immunoreactive neurons differed from those of another calcium-binding protein, parvalbumin, in the human amygdala (Sorvari et al. [1995] J. Comp. Neurol. 360:185-212). This suggests that these two calcium-binding proteins are located in different populations of neurons. PMID- 8726995 TI - Reticular thalamic region in the rabbit: organisation of efferents to the superior colliculus. AB - Although it is well-established that the reticular thalamic nucleus provides a strong GABAergic input to the dorsal thalamus, the existence of reticular efferents to other subcortical centres is less certain. In this study, we investigate whether the reticular nucleus projects to a major brainstem centre, the superior colliculus. The neuronal tracer, biotinylated dextran, was injected into superficial and deep layers of the superior colliculus of rabbits and the resultant labelling in the reticular region was examined. After large injections, which encompassed both superficial and deep collicular layers, two discrete populations of retrogradely labelled cells are seen in the region of the reticular nucleus. One population of retrogradely labelled cells lies in the dorsocaudal regions of the reticular nucleus, the classically defined visual sector. This group of retrogradely labelled reticular cells is also seen after injections into the superficial layers of the superior colliculus, but not after injections limited to the deeper collicular layers. The other population lies close to the ventromedial edge of the main body of the reticular nucleus, within a region referred to as the inner small-celled region. This group of small cells has been commonly thought to be part of the reticular nucleus, but our immunohistochemical studies suggest that this is a clearly separate region, a region continuous ventrally with zona incerta. The retrogradely labelled cells in the inner small-celled region are seen also after injections limited to the deeper collicular layers, but not after injections limited to the superficial collicular layers. Our results suggest functional heterogeneity within the reticular nucleus: Specifically, it suggests that the nucleus is in a position to influence the processing of visual information at both the dorsal thalamic and midbrain levels. PMID- 8726996 TI - Distribution of choline acetyltransferase and acetylcholinesterase in vocal control nuclei of the budgerigar (Melopsittacus undulatus). AB - The present study used histochemical methods to map the distributions of choline acetyl transferase (ChAT) and acetylcholinesterase (AChE) in the vocal control nuclei of a psittacine, the budgerigar (Melopsittacus undulatus). The distributions of ChAT and AChE in budgerigars appeared similar to that in oscine songbirds despite evidence that these systems have evolved independently. The magnicellular nucleus of the lobus parolfactorius in budgerigars, like the area X in songbirds, contained many ChAT labeled somata, fibers, and varicosities and stained densely for AChE. In contrast, the robust nucleus of the archistriatum (RA) and the supralaminar area of the frontal neostriatum in budgerigars, like the RA and the magnicellular nucleus of the neostriatum (MAN) in songbirds, respectively, contained few or no ChAT labeled somata, fibers, and varicosities and stained lightly for AChE. The central nucleus of the lateral neostriatum in budgerigars, like the higher vocal center (HVC) in songbirds, contained no ChAT labeled somata, moderate densities of ChAT labeled fibers and varicosities, and moderate levels of AChE staining. Two nuclei, the oval nucleus of the hyperstriatum ventrale (HVo) and the oval nucleus of the anterior neostriatum (NAo), contained no ChAT labeled somata, dense ChAT labeled fibers and varicosities, and moderate to high levels of AChE staining. The HVo and the NAo have no counterparts in songbirds but may be important vocal control nuclei in the budgerigar. Cholinergic enzymes are also described in other regions which may be involved in budgerigar vocal behavior, including the basal forebrain, the torus semicircularis, and the hypoglossal nuclei (nXII). PMID- 8726997 TI - White matter of the cerebellum of the chicken (Gallus domesticus): a quantitative light and electron microscopic analysis of myelinated fibers and fiber compartments. AB - Low magnification light microscopic examination of the white matter in appropriately stained avian and mammalian cerebellum reveals a mediolateral succession in which areas of large, heavily myelinated fibers alternate with areas containing nearly exclusively small fibers. A large fiber accumulation (LFA) and its medially adjoining small fiber area (SFA) form a fiber compartment, which, with related parts of cortex and central nuclei, constitutes a so-called cerebellar module. The composition and the apparent mediolateral heterogeneity of cerebellar fiber compartments was quantified in the chicken by morphometrical analysis of myelinated fiber profiles in light (LM) and electron (EM) microscopic micrographs. In LM versus EM, approximately 37% of the myelinated fiber population is neglected. This deficit concerns profiles that are smaller than 1.2 micron2 (diameter < 1.2 microns). EM analysis is therefore considered a prerequisite and forms the main part of this study. The myelinated fiber population has a left-skewed log normal size distribution. Ninety-nine percent of the myelinated fibers fall within the range of 0.1 to 20 microns2 (diameter = 0.4 5.0 microns) and 90% are even smaller than 7 micron2 (diameter < 3.0 microns). Small fibers are abundant in both parts of the compartment. Statistical comparisons provide quantitative confirmation of the LM distinction of LFAs and SFAs. It appears, moreover, that, apart from typical LFAs and SFAs, transitional zones rather than sharp borders can be distinguished between the two. The medial border of the LFA appears to be more sharply defined than its lateral border. Distinct mediolateral fluctuations were found with respect to fiber density (166 243 fibers/1,000 microns2), mean profile area (2.4-4.0 microns2), and interspace (31-47%). These differences reflect the contrast between LFA (lower density, larger mean profile area) and SFA (higher density, smaller mean profile area). The interspace discriminates less well between LFA and SFA but is often smaller in the LFA and larger in the SFA. The presented quantitative characteristics of mediolateral heterogeneity in the cerebellar fiber layer can be used as reference for morphometric studies on the different fiber systems of the cerebellar white matter and the functional organization of the compartments. PMID- 8726998 TI - Neural circuitry controlling vasopressin-stimulated scent marking in Syrian hamsters (Mesocricetus auratus). AB - In Syrian hamsters, arginine vasopressin (AVP) plays a critical role in the control of a form of scent marking called flank marking. Microinjection of AVP into the medial preoptic-anterior hypothalamus (MPOA-AH), lateral septal nucleus (LS), bed nucleus of the stria terminalis (BNST), and the periaqueductal gray (PAG) stimulates high levels of flank marking. Microinjection of an antagonist of the V1a-AVP receptor into sites such as the MPOA-AH inhibits expression of flank marking. The purpose of the present study was to investigate the neural circuit controlling flank marking by localizing the induction of Fos protein in response to the microinjection of AVP, a V1a-AVP antagonist (AVPA) or saline into the MPOA AH. Immediately after microinjection, hamsters were placed in a clean cage and their behavior was videotaped for 10 minutes. Ninety minutes after the behavioral experiment hamsters were perfused and their brains removed for subsequent immunocytochemical localization of Fos protein. The number of Fos-positive neurons was significantly greater in the BNST, PAG, and central amygdala (Ce) following the microinjection of AVP than following the microinjection of either AVPA or saline. In AVP-injected animals, the number of Fos-labeled cells in the Ce, PVN, and PAG increased with increased frequency of either flank marking or flank gland grooming. These data support the hypothesis that neurons within the MPOA-AH, BNST, and PAG play an important role in the control of flank marking and suggest that the Ce may be a previously unrecognized part of this neural circuit. PMID- 8726999 TI - Rapid down-regulation of tyrosine hydroxylase expression in the olfactory bulb of naris-occluded adult rats. AB - In most sensory systems, afferent innervation regulates morphological and biochemical characteristics of target cells for a limited time during development. Sensory deprivation experiments in adult rats also have suggested a critical period for afferent influences on olfactory bulb structure and function. Previous odorant deprivation studies that employed unilateral naris closure in neonatal rats demonstrated down-regulation of the catecholamine biosynthetic enzyme tyrosine hydroxylase (TH) in dopamine neurons intrinsic to the olfactory bulb. Accompanying the altered biochemical parameters was a decrease in bulb size. To distinguish between deprivation-induced alterations in TH expression secondary to developmental sequelae and those occurring in mature neurons, the consequences of unilateral naris closure were assessed in young adult rats. In agreement with previous studies significant postnatal increases occurred in TH expression and total protein, an indication of bulb size. At 30 days post closure, total protein was unaltered in the ipsilateral olfactory bulb but showed a small (12.9%), significant decline at 60 days. In contrast to the limited morphological consequences of odor deprivation, profound reductions occurred in TH expression. TH activity ipsilateral to the closure decreased significantly by 14 days post-closure and remained depressed for up to 6 months. In parallel with enzyme activity, TH immunoreactivity did not decline in the first few days post closure. In situ hybridization revealed that TH mRNA levels decreased rapidly, i.e., by 2 days post-closure, reached a nadir at 1 month, and remained depressed for at least 6 months. The capacity of odor deprivation in the adult rat olfactory system to down-regulate TH expression suggests that this phenotypic alteration occurs independently of a presumed critical period. PMID- 8727000 TI - Neural connections in and around the cavernous sinus in rat, with special reference to cerebrovascular innervation. AB - There is a confluence in and around the cavernous sinus of neural pathways innervating the intracranial structures. To determine the patterns of innervation, particularly of the cerebral arteries, we stained whole-mount preparations of the cavernous sinus and adjacent regions of the rat for acetylcholinesterase. The cavernous nerve plexus, with several small ganglia, mainly occupied the lateral wall of the sinus and extended laterally above the ophthalmic and maxillary divisions of the trigeminal nerve, in relation to the oculomotor and trochlear nerves. The cavernous plexus was connected to the pterygopalatine ganglion, the trigeminal ganglion, and the abducens nerve. The elongated pterygopalatine ganglion consisted of an orbital part, from which parasympathetic fibers ran to the cerebral arteries, and a cavernous part. Nerves from the lateral extension of the cavernous plexus ran rostrally into the orbit along the oculomotor, trochlear, and ophthalmic nerves, and caudally to the pineal gland along the trochlear nerve. Several branches also ran over the dura mater. Caudal to the cavernous sinus, we found two large nerves and a number of small nerves that ran between the nerves surrounding the internal carotid artery and the abducens nerve. These nerves may represent additional parasympathetic and/or sensory pathways to the cerebral arteries. PMID- 8727001 TI - Anatomical basis of a congenital hearing impairment: basilar papilla dysplasia in the Belgian Waterslager canary. AB - Recent investigations into the anatomy of the inner ear of Belgian Waterslager canaries (BWC) have demonstrated myriad malformations associated with dysgenesis of the pars inferior of the otocyst. In those studies, the surface anatomy of BWC's basilar papilla and sacculus was examined utilizing scanning electron microscopy. In the present investigation, we utilized both light microscopy and transmission electron microscopy to describe the cross sectional anatomical pathology of the BWC's basilar papilla. Examination of the BWC's organ of Corti revealed numerous dysmorphologies: 1) hair cells from the tall hair cell region appeared broad and stunted, with deformed cuticular plates, abnormal stereocilia, and recognizable microvilli; 2) quantitative analysis of these hair cells revealed disproportionately large nuclei and abnormally short stereocilia; 3) hair cells from the short hair cell region of the papilla were absent, replaced by a population of large cells with electron-lucent cytoplasm; and 4) the tectorial membrane in the BWC papilla was narrow, covering only the area where the deformed tall hair cells were found. The malformations appeared to be more severe at the apex and midsection of the basilar papilla than at the base. These observations allow us to suggest a hypothesis to account for the distinct anatomofunctional hearing deficit observed in these birds. In addition, they further support our hypothesis that the inner ear of BWC is afflicted by a disorder similar to Scheibe's dysplasia, the most common inner ear defect associated with congenital hearing loss in humans. PMID- 8727002 TI - Efferent projections of the anterior perirhinal cortex in the rat. AB - Because convulsive seizures develop very rapidly from kindling sites in the anterior perirhinal cortex, we studied perirhinal efferents by using the anterograde tracer Phaseolus vulgaris leucoagglutinin (PhAL). PhAL injections into the anterior perirhinal cortex labelled a prominent network of fibers within the frontal cortex that was most dense within layers I and II and layer VI. As individual PhAL injection sites within the perirhinal cortex were restricted to one or two adjacent laminae, we were able to determine that layer V was the main source of the perirhinofrontal projection. This was confirmed by frontal cortex injections of the retrograde tracer Fluorogold (FG). Other cortical areas with densely labelled fibers following perirhinal PhAL injections included the agranular insular, infralimbic, orbital, parietal, and entorhinal cortices. Moderate to mild fiber labelling was also noted in the posterior piriform, temporal and occipital cortices, and the claustrum. Subcortical labelling was seen in the nucleus accumbens; fundus striati; basal and lateral amygdala nuclei; the "acoustic thalamus"; and the central grey. Several of these cortical and subcortical projections were bilateral. The different laminar origin of these perirhinal efferents is discussed. These results confirmed our prediction of extensive direct projections from the anterior perirhinal cortex to the frontal cortex in the rat. The significance of this projection is discussed with special reference to the anatomical basis of convulsive limbic seizures. PMID- 8727003 TI - Temporal and spatial pattern of MASH-1 expression in the developing rat retina demonstrates progenitor cell heterogeneity. AB - The temporal and spatial pattern of mammalian achaete-scute homolog 1 (MASH-1) expression in the developing rat retina was examined in an effort to correlate achaete-scute homolog expression with the generation of particular cell classes. The expression of MASH-1 was restricted to the latter portion of retinal neurogenesis and was most closely correlated with the appearance of bipolar cells and Muller glia, two cell classes that are generated late in retinogenesis. We also examined the proliferative nature of the MASH-1 -expressing cell type to confirm that MASH-1 is expressed by progenitor cells and to determine the proportion of the proliferating population that expresses MASH-1. MASH-1 was expressed by only 10-30% of the total proliferating population, depending on the age examined. Thus, MASH-1 expression provides a molecular marker of heterogeneity among retinal progenitor cells and may play a role in the commitment and/or differentiation of one or more of the late-appearing retinal phenotypes. PMID- 8727004 TI - Use of mammalian liver cells for artificial liver support. AB - Advances in orthotopic liver transplantation have improved the survival rate of both acute and chronic liver failure patients to nearly 70%. However, the success of this treatment modality has created an international organ shortage. Many patients die while awaiting transplantation in part due to the minimal capacity to store viable transplantable livers beyond 24 h. Additionally, for many areas of the world, routine use of whole liver transplantation to treat liver disease is impractical due to the demands on both financial and technical resources. Potentially, these issues may be alleviated, at least in part, by the use of liver cell transplantation or cellular-based liver assist devices. The well documented regenerative capacity of the liver may obviate the need for whole organ transplantation in some instances of acute failure, if the patient may be provided temporary metabolic support. Although other patients ultimately may require transplantation, a longer period of time to find a suitable organ for transplantation may be gained by that supportive therapy. The field of liver cell transplantation may offer solutions to patients with inherited metabolic deficiencies or chronic liver disease. The potential to treat an hepatic disorder by using only a fraction of the whole liver would increase the number of whole organs available for orthotopic liver transplantation. Research in the fields of hepatocyte based intra- and extra-corporeal liver support is providing evidence that these therapeutic modalities may ultimately become routine in the treatment of severe liver disease. A historic overview of that technology along with its current status is discussed. PMID- 8727005 TI - Significant improvement of survival by intrasplenic hepatocyte transplantation in totally hepatectomized rats. AB - The effect of intrasplenic hepatocyte transplantation (HTX) was studied in an experimental model of acute liver failure in rats with chronic liver atrophy. Rats underwent a portacaval shunt operation on Day -14 to induce liver atrophy, and underwent total hepatectomy on Day 0 as a start of acute liver failure. Intrasplenic hepatocyte or sham transplantation was performed on Day -7,-3, or -1 (n = 4 to 6 per group). During the period following hepatectomy, mean arterial blood pressure was maintained above 80 mm Hg and hypoglycaemia was prevented. Severity of hepatic encephalopathy was assessed by clinical grading and EEG spectral analysis, together with determination of blood ammonia and plasma amino acid concentrations, and "survival" time. Histological examination of the spleen and lungs was performed after sacrifice. Intrasplenic hepatocyte transplantation resulted in a significant improvement in clinical grading in all transplanted groups (p < 0.05), whereas a significant improvement in EEG left index was seen only in the group with transplantation on Day -1 (p < 0.05). In contrast to hepatocyte transplantation 1 day before total hepatectomy, rats with hepatocyte transplantation 3 and 7 days before total hepatectomy showed a significant 3- and 2-fold increase in "survival" time compared to sham transplanted controls: HTX at Day -1: 7.5 +/- 0.3 h vs. 5.9 +/- 0.6 h (p > 0.05), HTX at Day -3: 19.7 +/- 3.7 h vs. 6.5 +/- 0.3 h (p < 0.05), and HTX at Day -7: 13.8 +/- 3.2 h vs. 6.3 +/- 0.3 h (p < 0.05). Furthermore, rats with hepatocyte transplantation on Day -3 and -7 showed significantly lower blood ammonia concentrations after total hepatectomy (p < 0.0001). Histological examination of the spleens after sacrifice showed clusters of hepatocytes in the red pulp. Hepatocytes present in the spleen for 3 and 7 days showed bile accumulation and spots of beginning necrosis. The present data show that in a hard model of complete liver failure in portacaval shunted rats, intrasplenic hepatocyte transplantation is able to prolong "survival" time significantly 2- to 3-fold. The relevance of this observation for human application is discussed. PMID- 8727006 TI - Blood stem cell collection: factors influencing the recovery of granulocyte macrophage colony forming cells. AB - We evaluated data from all blood cell (BC) collections performed in our institution between 1989 and 1995 to determine factors influencing the outcome of collection. One hundred and thirty-three collections were performed on 106 patients. Malignant diagnoses were: non-Hodgkins lymphoma (NHL) in 35%, multiple myeloma in 31%, breast cancer in 26%, and Hodgkin's disease in 8%. Collections were obtained routinely in myeloma and breast cancer and due to bone marrow involvement with malignancy or inaspirable bone marrow in lymphoma patients. Collections were obtained on a Cobe Spectra or Baxter-Fenwall CS3000+. Engraftment potential was determined by methylcellulose colony assay (CFU-GM), with a target of > 10 x 10(4) CFU-GM/kg. Apheresis nucleated cell count correlated significantly, albeit weakly (r = 0.26), with CFU-GM with a cell count of > 5 x 10(8)/kg resulting in an adequate number of CFU-GM in 78% of patients. In univariant analysis outcome of collection was significantly influenced by the patients age (p = 0.01), malignant diagnosis (p < 0.001), reason for collection (p = 0.002), and the mobilization regimen (p = 0.01). The nature of the apheresis device used did not influence outcome. Only malignant diagnosis was significant (p < 0.001) in multivariate analysis. We conclude that the outcome of BC is most strongly influenced by patient factors such as malignant diagnosis. These factors must be considered when comparing the outcome of different mobilization regimens and when planning collection strategies. PMID- 8727007 TI - Correction of Fanconi anemia type C phenotypic abnormalities using a clinically suitable retroviral vector infection protocol. AB - Fanconi anemia (FA) is a complex autosomal recessive disease with hematologic manifestations characterized by a progressive hypoplastic anemia, hypersensitivity to clastogenic agents, and an increased incidence of acute myelogenous leukemia. The cDNA that corrects one of four FA complementation subtypes, named Fanconi anemia Type C (FAC) has recently been identified. We constructed a a simplified recombinant retrovirus (vMFGFAC) encoding only the FAC cDNA, and tested its ability to correct the FAC defect in a lymphocytic cell line and primary mobilized blood progenitor cells. In addition, the gene transfer efficiency using a clinically applicable gene transfer protocol into normal primitive hematopoietic progenitor cells, high proliferating potential colony forming cells (HPP-CFC), derived from CD34+ purified cord blood cells was examined. The gene transfer efficiency was significantly enhanced when cells were transduced with supernatant while adherent to a 30/35 KD fragment of fibronectin, FN30/35, and was similar to efficiency obtained by coculture with retrovirus packaging cells. Transduction of an FAC deficient lymphoid cell line with vMFGFAC supernatant resulted in an enhanced cell viability, and G-CSF mobilized peripheral blood cells from an FAC-deficient patient transduced with the vMFGFAC virus demonstrated enhanced progenitor cell colony formation. These data indicate that the vMFGFAC virus allows functional complementation of FAC in lymphoblasts and primary hematopoietic progenitors, and that primitive cord blood hematopoietic stem/progenitor cells can be transduced at an efficiency comparable to protocols using cocultivation if adherent to FN 30/35 fragment. PMID- 8727008 TI - Bulk cryopreservation of isolated islets of Langerhans. AB - Current methods to isolate human islets of Langerhans are limited and multiple donors are required for successful reversal of longstanding Type 1 diabetes mellitus. Cryopreservation of isolated islets is an effective method of storing and pooling islets. Current cryopreservation protocols are cumbersome due to current practices of placing small aliquots of islets per individual freezer tube. In the present study, we examined the application of a blood freezer bag for the cryopreservation of isolated islets by slow cooling and rapid thawing. Freezing and thawing profiles generated using thermocouples placed inside a 500 mL Cryocyte (Baxter) blood freezer bag showed that a longer equilibration period at -7.4 degrees C was necessary to consistently achieve nucleation and cooling profiles similar to those observed in glass tubes. When known numbers of rat islets were placed in the freezer bag and the cryoprotectant dimethyl sulfoxide (DMSO) was added in a stepwise fashion and removed using a sucrose dilution, the islet recovery compared with glass tubes was 92 +/- 4.8 vs. 90 +/- 2.3% (n = 4, p = ns, Mann-Whitney U-test). When purified canine islets were cryopreserved in a single freezer bag or in multiple glass tubes, the recovery was similar (78.8 +/- 12.5% recovery for freezer bag vs. 82.3 +/- 5.3% for glass tubes; n = 6, p = ns). In vitro function was equivalent for both groups. The stimulation index of insulin release during glucose perifusion (stimulated over basal insulin secretion) for canine islets cryopreserved in a freezer bag vs. glass tubes was 3.2 +/- 1.0 and 2.3 +/- 1.3, respectively (n = 6, p = ns). These values were significantly lower than the nonfrozen control islets (6.9 +/- 2.4, p < 0.05). When 2000 canine islets cryopreserved in either a freezer bag, or glass tubes were transplanted into diabetic nude mice, the animals became and remained normoglycemic posttransplant. We conclude that the survival of freshly isolated canine islets cryopreserved in a single freezer bag is equivalent to the glass tube method. Bulk cryopreservation of islets in a single freezer bag will facilitate effective low temperature tissue banking to support ongoing clinical trials of islet transplantation. PMID- 8727009 TI - A simple method for cardiac surgery in rats. AB - In our laboratory we have developed a relatively simple method for cardiac surgery in rats. The operation is carried out through a small incision in the chest wall using inexpensive equipment. This method allows for the delivery of tissue fragments and cells from a donor rat to an intact or damaged area of ventricular myocardium of a host rat, with easy subsequent localisation of the transplanted/grafted tissue. The rats recover well after the surgery and survive for long periods of time. The technique could also be used for the direct injection of chemicals or molecular probes into the heart. In our experiments we have found that embryonic rat cardiomyocytes that have been transplanted into adult host rat ventricular myocardium using this method survive and develop characteristics typical of heart muscle, thus indicating that using this technique the host heart offers a favourable environment for the transplanted embryonic heart cells. PMID- 8727010 TI - Long-term expression of a fluorescent reporter gene via direct injection of plasmid vector into mouse skeletal muscle: comparison of human creatine kinase and CMV promoter expression levels in vivo. AB - Expression of a fluorescent reporter gene has been studied using two alternate promoters to transcribe the green fluorescent protein (gfp) from Aequorea victoria. The human cytomegalovirus (CMV) enhancer/ promoter or the human muscle specific creatine kinase promoter (CKM) were inserted along with the gfp cDNA into a plasmid expression vector based on a modified adeno-associated virus genome. Naked plasmid DNA was injected into the hamstring muscle of mdx mice and gfp gene expression determined from frozen muscle sections taken at 4, 14, and 42 days postinjection. Fluorescence patterns obtained by photomicroscopy and quantitative fluorescence measurements indicated a near-linear increase in the accumulation of the gfp in skeletal muscle during the length of the study, with gfp expression at 42 days being roughly four times the values obtained at 4 days. The levels of expression of gfp from the CKM construct were consistantly higher than for the CMV construct. The CKM promoter/expression vector combination demonstrates significant potential for simple, direct delivery and long-term, high-level expression of genes in skeletal muscle. PMID- 8727011 TI - A potential alternative strategy for myoblast transfer therapy: the use of sliced muscle grafts. AB - Excellent long-term survival (up to 1 yr) of donor skeletal muscle cells was demonstrated using a mouse Y-chromosome specific probe, following the transplantation of grafts of whole muscles from male "normal" C57B1/10Sn mice into dystrophic muscles of female host mice. After the transplantation of equivalent sliced muscle grafts there was extensive movement of the male donor cells and fusion with host myofibres. This contrasts with the extremely poor survival of isolated myoblasts after injection into the same mouse model for Duchenne muscular dystrophy. The use of sliced muscle grafts may therefore represent a potential alternative approach to myoblast transfer therapy. PMID- 8727012 TI - Commentary on the present state of knowledge for myoblast transfer therapy. PMID- 8727013 TI - Selective venous clamping in rat hepatocyte transplantation. PMID- 8727014 TI - The effects of muscle fatigue on and the relationship of arm dominance to shoulder proprioception. AB - It is hypothesized that proprioceptive information plays an important role in joint stabilization and that muscle fatigue may alter proprioceptive ability. The purpose of this study was to determine what effect shoulder muscle fatigue has on glenohumeral proprioception and to examine the relationship between arm dominance and shoulder proprioception. Eighty subjects without a history of glenohumeral pathology participated. Each was seated on an isokinetic dynamometer with a randomly selected shoulder positioned in 90 degrees of abduction and elbow flexion. With vision blinded, the arm was passively positioned in 75 degrees of external rotation for 10 seconds, then passively returned to the neutral starting position. Three trials each of active and passive repositioning (2 degrees/sec) were recorded. Following a fatigue protocol, both active and passive repositioning were reassessed. Testing order was randomized. A significant difference was detected between pre- and post-fatigue scores. No significant difference was detected between dominant and nondominant extremities. No relationship between arm dominance and shoulder proprioception was established. It is concluded that shoulder proprioception is diminished in the presence of shoulder muscle fatigue, suggesting clinical rehabilitation protocols must emphasize increasing muscular endurance. PMID- 8727015 TI - Clinical measurement of head and shoulder posture variables. AB - A reliable method that is practical for clinical use for measuring head posture in standing has not been demonstrated in the literature. The purposes of this study were: 1) to produce a reliable method for measuring sagittal plane head and shoulder posture that is practical for clinical use and 2) to perform a pilot study to compare means between a nonpatient sample and a sample of people who have a history of recurring cervical pain. A method of measuring sagittal plane postural alignment of the head and shoulder in relationship to the lateral malleolus was developed using a carpenter's tri-square with a line level attached to the horizontal arm and a goniometer with a line level attached to the horizontal arm. Horizontal measures were taken with the tri-square from a vertical reference point to the tragus of the ear, to the shoulder axis, and to the lateral malleolus. Angular measures were taken with the goniometer of C7 tragus with the horizontal and tragus-corner of the eye with the horizontal. A reliability study was performed by two therapists with 15 non-patient subjects. Intraclass correlation coefficients measuring interrater reliability for horizontal and angular measures were as follows: 1) tragus to lateral malleolus: R = .87; 2) shoulder to lateral malleolus: R = .91;3) tragus to shoulder: R = .7;4) angle tragus-eye-horizon: R = .68; and 5) angle C7-tragus-horizon: R = .34. In the pilot study, 41 nonpatient subjects (11 males and 30 females, 20-45 years) and 10 patient subjects with cervical pain (one male and nine females, 23-43 years) were measured. Means calculated for horizontal distances (in cm) were: 1) tragus to lateral malleolus (nonpatients: 8.14; patients: 6.65);2) shoulder to lateral malleolus (nonpatients: 5.47; patients: 5.98); and 3) tragus to shoulder (nonpatients: 2.78; patients: .56). Means calculated for angular measures were: 1) tragus-eye-horizon (nonpatients: 18.78 degrees; patients: 21.57 degrees) and 2) C7-tragus-horizon (nonpatients: 49.34 degrees; patients: 49.43 degrees). No statistically significant differences were found between the two samples in this pilot study using an independent two-sample t test. This study demonstrates a reliable and practical method for taking postural measurements in the clinic and describes a pilot study for comparing a patient and a nonpatient sample. PMID- 8727016 TI - The effect of an ankle orthosis on ankle range of motion and performance. AB - Ankle joint orthoses are used for rehabilitation and/or prevention of ankle sprains. The purpose of this study was to determine the effect of the Malleoloc ankle joint orthosis on active and passive range of motion reduction and on a jumping and a figure-eight running test. Twelve subjects with a history of inversion ankle sprain and documented increased anterior translation in a drawer test participated in the study. Active and passive range of motion for inversion was determined with and without the orthosis and pre- and post-exercise. Additionally, performance tests for figure-eight running and jumping were administered. The results showed that the tested orthosis 1) restricted the active range of motion and passive inversion substantially, 2) reduced the other movement degrees of freedom only minimally, 3) provided the same movement restriction before and after exercise, and 4) did not affect performance. The Malleoloc ankle joint orthoses can, therefore, restrict ankle joint motion without affecting performance negatively. PMID- 8727017 TI - Relationship between three static angles of the rearfoot and the pattern of rearfoot motion during walking. AB - The purpose of this study was to determine the relationship of the static angle of the rearfoot during single leg standing, relaxed standing foot posture, and subtalar joint neutral position with the pattern of rearfoot motion during walking. The authors felt that this study was important to gain a better understanding of the relationship between dynamic rearfoot motion and three static rearfoot angles which are often included in foot examination procedures. The pattern of rearfoot motion was assessed using two-dimensional video recordings for each lower extremity of 31 healthy young adult subjects with a mean age of 25.2 years. The mean path of rearfoot motion during walking crossed relaxed standing foot posture but did not cross single leg standing or subtalar neutral position. These findings suggest that the mean path of rearfoot motion during the first 60% of the walking cycle occurs between the static angles of relaxed standing foot posture and single leg standing. In addition, the static angle of the rearfoot in single leg standing may serve as a clinical indicator of the degree of maximum rearfoot eversion occurring during the walking cycle. PMID- 8727018 TI - Effects of aquatic therapy on joint flexibility and functional ability in subjects with rheumatic disease. AB - Currently, there is a deficit of objective data showing the efficacy of aquatic exercise therapy in improving function in individuals with rheumatic disease. The purpose of this study was to quantify the effectiveness of aquatic therapy in changing joint motion, functional status, assistance, pain, and difficulty in 13 subjects with rheumatic disease. Active joint motion and Functional Status Index scores of assistance, pain, and difficulty were taken prior to and following 8 weeks of aquatic therapy. Multivariate analysis of variance, analysis of covariance procedure controlling for pain, and a Pearson correlation between active joint motion and Functional Status Index changes and between assistance, pain, and difficulty scores of the Functional Status Index were conducted. Results showed a significant difference between the pre and post means of active joint motion, Functional Status Index, pain, and difficulty (p < .05); a significant, negative relationship between pain and active joint motion of select joints; and positive correlations between assistance and pain, assistance and difficulty, and pain and difficulty. The decreased pain and difficulty experienced in performing daily tasks contributed significantly (94%) to the overall increased functional status and active joint motion for select joints. These findings support aquatic therapy as an effective means of increasing joint flexibility and functional ability while reducing pain and difficulty with daily tasks. PMID- 8727019 TI - Controlled active mobilization following surgical repair of the avulsed radial attachment of the biceps brachii muscle: a case report. AB - Traumatic disruption of the radial attachment of the biceps brachii muscle may require surgical reattachment to obtain and maintain optimum anatomical relationships and ensure return to full, unlimited activity, particularly in the young athletic patient with high muscle performance demands. Postoperative evaluation and care following surgical reattachment of the avulsed radial tendon of the biceps brachii muscle has not previously been reported in the rehabilitation literature. The purpose of this case study is to report successful treatment of a gymnast using static splinting, controlled active mobilization of the elbow, and strengthening exercises following surgical reattachment of the avulsed radial attachment of the biceps brachii muscle. PMID- 8727020 TI - Dermatological problems in the athlete. AB - Sports activities expose the skin to a variety of risk factors that can affect an athlete's performance. The sports physical therapist must be able to identify and properly manage dermatological problems encountered by athletes. Common dermatological conditions resulting from mechanical, infectious, environmental, and sexual factors are presented. Discussion of etiology, signs and symptoms, treatment options, and preventative measures are intended to guide the physical therapist in making appropriate decisions regarding skin disorders. PMID- 8727022 TI - Technical and ethical problems associated with patch testing. AB - When relevant allergens are identified by patch testing, and patients can avoid them in their environment, improvement of dermatitis is the rule. Some forms of chronic dermatitis may not clear completely, but patients are usually satisfied with modification of their previously more severe problem. In 1981 when asked what he felt were the five most important advances in clinical dermatology during the 20th century, Dr. Marion B. Sulzburger, an icon in American dermatology, said that, "The increased use and usefulness of the patch test and the international standardization of test concentrations and methods" was number one. Those of us who are enthusiastic patch testers and fascinated by the evaluation of patients with irritant and allergic contact dermatitis would agree. It is a thrilling clinical experience to be able to tell a machinist that he need not stop his lifelong occupation, but instead will do fine if he will simply avoid the waterless hand cleanser he has been using which is preserved with glutaraldehyde. The woman whose facial dermatitis has embarrassed her for years and clears when she stops using the Quaternium-15 preserved moisturizer that you have identified in your patch testing is grateful to you forever, and again happy in her own life. In 1991, my research assistants, Patricia Norris and Mary Lou Belozer, and I studied 30 university hospital workers who answered our advertisement asking for individuals who believed they were troubled by their rubber gloves (unreported study). By evaluating these people through history, physical examination, and patch testing, we were able to prove glove relatedness in 14 of them. Nine of the 14 had contact urticaria to latex, and only 5 had allergic contact dermatitis to rubber glove ingredients. Fifteen of our patients had irritant dermatitis. In this study, none of the patients with allergic contact dermatitis to glove ingredients had contact urticaria. However, since that time, we have observed a number of patients who had both forms of allergic reaction. Three of our patients who presented with nummular (patchy) hand dermatitis also had contact urticaria to latex, but no positive patch tests. With latex glove avoidance, their dermatitis resolved; an example of how scratching urticaria can eventuate in longer lasting dermatitis in some people ("the itch that rashes"). The patients presented to their dermatologist with dermatitis, but their true initiating event was urticaria which lasted only hours. The gratifying part of this study was that patch testing and contact urticaria testing allowed us to discover the 5 patients with allergic contact dermatitis and the 9 patients with contact urticaria who could benefit from glove alternatives. We were also able to assure patients in the remainder of the group that their hand eczema was not glove induced, but rather was related to their wet work. In most instances, therapeutic intervention helped, but in several cases job changes were required. Patch testing, when done properly, produces exciting results. When done improperly, it confuses and misleads patients and results in embarrassment to physicians who cannot properly interpret their results. Should a physician choose to include patch testing in his or her evaluation of patients with contact dermatitis it is essential, in my view, that he or she have highly developed skills in the diagnosis and treatment of skin diseases, and that these physicians be elaborately trained in the techniques of application and the methods of interpretation of patch tests. PMID- 8727023 TI - Contact allergy to corticosteroids. Diagnosis and management. PMID- 8727021 TI - Contact dermatitis. Clinical perspectives and basic mechanisms. PMID- 8727024 TI - Aeroallergen contact dermatitis. PMID- 8727025 TI - The spectrum of cutaneous patch-test reactions in patients with atopic dermatitis. AB - We have outlined rationale and problems associated with three types of patch tests that may provide diagnostic help in patients with difficult-to-manage AD. Each of these methods is imperfect and in need of greater study using a critical approach, better techniques, more attention to irritant controls, and better definition of atopic populations. Proof of validity for aeroallergen patch testing will necessarily require large quantities of purified antigens. In spite of all these problems, patch testing can be useful and sometimes essential for the diagnosis and management of difficult AD. PMID- 8727026 TI - Tissue-arterial PCO2 difference is a better marker of ischemia than intramural pH (pHi) or arterial pH-pHi difference. AB - Gastric intramucosal pH (pHi) is often calculated by the Henderson-Hasselbalch equation, using arterial plasma [HCO3-]ap and PCO2 measured in saline obtained from a silastic balloon tonometer after equilibration in the lumen of the stomach. A pHi value less than approximately 7.3 pH units is often taken as evidence of intestinal ischemia. An alternative measure is tissue PCO2 (PtCO2) PaCO2 difference [P(t-a)CO2]. The idea is that PtCO2 will increase slightly relative to PaCO2 as O2 supply decreases, and then increase strikingly when flow decreases to a critical value, because of liberation of CO2 from tissue Hco3- by anaerobically generated strong acid. A third method is arterial plasma pH (pHap) pHi difference [pH(ap-i)]. We used mathematical simulations to test the hypotheses that calculated pHi is independent of arterial acid-base status; and pH(ap-i) provides the same information as does P(t-a) CO2. Using the Van Slyke version of the arterial whole blood [standard base excess] ([SBE]aWB) equation, it was found that a change in [SBE]aWB at constant PaCO2 and constant PtCO2 produces a change in calculated pHi (P = 0), such that the relation between changing [SBE]aWB and changing pHi is predictable by a single polyomial equation (R2 = .999). pH(ap-i) avoids this confounding influence of [SBE]aWB. However, it was further shown that pH(ap-i) can be associated with a wide range of P(t-a)CO2, depending on the magnitude of pH(ap-i), and on the PaCO2 at which P(t-a)CO2 is measured. We conclude that P(t-a)CO2 is a more reliable index of gastric oxygenation than is pHi alone or pH(ap-i). PMID- 8727027 TI - Effects of continuous positive airway pressure on cardiac output and plasma norepinephrine in sedated pigs. AB - Continuous positive airway pressure (CPAP) increases cardiac output (CO) in congestive heart failure (CHF). In six sedated pigs that were normovolemic (NV) and hypervolemic (HV), and seven previously instrumented pigs with pacing-induced CHF, we tested the hypothesis that this is associated with decreased total body sympathetic nerve activity (SNA). Hemodynamic variables and plasma norepinephrine level measurements were measured at baseline, CPAP 5 and 10 cm H2O, and recovery. Arterial O2 saturation was maintained at greater than or equal to 90% and PCO2 did not change. For NV baseline plasma norepinephrine level (PNE) was 97 +/- 61 pg/mL, CO 2.4 +/- .5 L/min, and pulmonary wedge pressure (Pw) 10.1 +/- 2.4 mmHg and did not change with CPAP. HV and CHF were associated with increased baseline Pw (18-21 mmHg). Baseline CO was increased with HV and unchanged with CHF. Baseline PNE was increased 4 to 5-fold with both HV and CHF. CO increased at CPAP 5 compared with baseline with both HV and CHF. However, PNE decreased with CPAP in HV, and increased with CPAP in CHF. Increased CO was always associated with decreased systemic vascular resistance. We conclude the following: (1) increased CO with CPAP can be associated with either increasing or decreasing SNA; (2) CPAP can produce increases in CO when the heart is distended whether baseline LV function is relatively normal (HV) or depressed (CHF); and (3) there are probably a number of different mechanism increasing CO with CPAP and these may vary from condition to condition. PMID- 8727028 TI - Dynamic right and left ventricular interactions in the rabbit: simultaneous measurement of ventricular pressure-volume loops. AB - PURPOSE: This study was performed to characterize the dynamic factors determining ventricular interdependence in an open-pericardium intact animal model. MATERIALS AND METHODS: Simultaneous measures of right ventricular (RV) and left ventricular (LV) pressures and volumes in 6 urethane-anesthetized open-chested, open pericardium rabbits. RV and LV V were calculated every 2 milliseconds. Measurements were made at initial baseline blood volume, and again after two infusions of 20 mL/kg isoconductive colloid solution. At each blood volume level, partial aortic (AO), pulmonary artery (PAO), and inferior vena caval (IVC) occlusions were performed. Biventricular diastolic compliance and end-systolic elastance were calculated from these data. RESULTS: Baseline end-diastolic (ED) and end-systolic (ES) V were 3.29 +/- 0.55 and 2.43 +/- 0.33 mL (mean +/- SD) for the LV, and 3.38 +/- 1.56 and 2.84 +/- 1.36 mL for the RV, respectively. AO increased all LV pressure and volume (P < .05) but did not alter RV ED volume (2.85 +/- 1.20 mL) or ED pressure (3.3 +/- 2.0 to 3.6 +/- 2.1 mm Hg). PAO increased RV ES pressure (P < .05) but did not alter RV ED volume, ED pressure, or ES volume, although it decreased LV ED volume (2.82 +/- 0.59, P < .05). AO also immediately increased end-systolic RV elastance to a value greater than that defined by IVC (7.9 +/- 4.4 to 10.9 +/- 6.6 mm Hg/mL, P < .05). Intravascular volume expansion though increasing baseline pressure and volume, did not alter qualitatively biventricular responses to AO, PA, or IVC. CONCLUSION: Ventricular interdependence has both systolic and diastolic components that have differing directional effects. In the pericardectomized rabbit, increases in RV ED volume decrease LV ED volume by decreasing LV diastolic compliance, but do not alter LV systolic function. Whereas, increases in LV ED volume decrease RV ES volume resulting in an increase in RV maximal elastance, but minimally alter RV diastolic function. PMID- 8727029 TI - Applications of molecular biology and biotechnology: antibody therapy of sepsis. AB - The use of antibody therapy for the treatment of infections and inflammatory disease is well established. Unfortunately, clinical studies of antiendotoxin and anti-TNF monoclonal antibodies have failed to show clear physiological or survival benefit. Little information is available regarding the effect of antibodies to cytokines other than TNF in human sepsis. Limited pre-clinical data indicate that IL-6 antibodies may abrogate the effects of endotoxin infusion, but no human studies have been performed. Although both monoclonal and polyclonal antibodies have the potential to protect septic humans, at this time it is the polyclonal antibodies that have shown the greatest promise. Each type of antibody possesses specific advantages and limitations, the ultimate effectiveness of which will need to be proven in large randomized clinical trials. PMID- 8727030 TI - Organ-specific therapy in critical illness: interfacing molecular mechanisms with physiological interventions. AB - Sepsis and SIRS is the outward manifestation of a generalized uncontrolled inflammatory response, which, if sustained, induces widespread endothelial damage and MODS. Immunomodulating therapies, at present, have proven ineffective in reducing morbidity and mortality, presumably because of the heterogeneous nature of sepsis and septic shock and the reciprocating and redundant nature of this inflammatory cascade. Organ-specific therapies can support life but impair both organ-specific function and remote organ function. Novel therapies aimed at minimizing further organ dysfunction may improve outcome in a cost-effective fashion by preventing both further primary organ dysfunction or remote organ dysfunction secondary to the subsequent activation of the inflammatory response. PMID- 8727031 TI - External reality: the elusive dimension of psychoanalysis. AB - Current psychoanalytic thinking reflects important differences concerning external reality. It is absent from the theories of the hermeneuticists, and it is excessively present in self psychology. Those treating victims of "real" trauma frequently believe that psychoanalysis is not appropriate for their patients. This has led to the unacceptable separation of individuals: those who are traumatized and those who are neurotic. I believe that a review of the role of external reality in psychic functioning is overdue, and the present work is an attempt to reassess our knowledge of this important dimension and to find its proper place in psychoanalysis. PMID- 8727032 TI - Character portrayal in Anthony Trollope's Barset novels: a psychoanalytic appraisal. AB - Anthony Trollope's novels enjoyed great popularity in his own day, and the past several decades have seen a considerable revival of interest in them. His most popular series, the six Barset novels, exemplify his love of life, his psychological sensitivity, and his profound sense of the meaning and value of the smallest nuances of human behavior and interaction. In his comprehension of the psychological vicissitudes of ordinary daily life he has been compared to Chaucer. This paper studies a portrait of a severely depressed character in Trollope's own favorite work, The Last Chronicle of Barset. PMID- 8727033 TI - The role of the other in mourning. AB - Mourning is a highly social process. This paper's thesis is that surviving objects play a crucial role in the accomplishment of a number of the major psychological tasks of mourning. To illustrate the thesis, the psychoanalytic treatment of a man suffering the aftermath of parental loss in adolescence is presented. The failure of the social surround to assist with mourning is identified and explored, and the treatment implications of the role of the other in mourning are discussed. PMID- 8727034 TI - Analyst and patient at midlife. AB - Increasingly it seems that analysts and their patients are in the midlife period. If analyst and patient are both going through a midlife crisis involving similar issues, this may have repercussions in the analysis. Turning to the literature for guidance reveals a paucity of papers concerning the clinical application of midlife developmental theory. This article explores such an application by describing the effect of two midlife changes-time urgency and role reversal with respect to aging parents-on the transference-countertransference in the analyses of two middle-aged patients. PMID- 8727035 TI - A note on empathy and the analyst's transference. PMID- 8727036 TI - "Dim spot": a variant in between countertransferential "bright" and "blind" spots. PMID- 8727037 TI - Reproductive conflicts in incest victims: an unnoticed consequence of childhood sexual abuse. PMID- 8727038 TI - A presence announced by its absence. PMID- 8727039 TI - Rat islet cell lines produced by retroviral transduction of SV40 T antigen. PMID- 8727040 TI - Representativeness of microorgans from human colorectal tumors. PMID- 8727041 TI - Reconstituted basement membrane (Matrigel): a useful semisolid medium for growth of tumor cell colonies. PMID- 8727042 TI - Non-alpha-fetoprotein-producing anaplastic hepatoblastoma cell line. PMID- 8727043 TI - Mechanistics of formation and ultrastructural evaluation of hepatocyte spheroids. AB - Freshly harvested rat hepatocytes form spheroids on uncoated positively charged polystyrene surfaces. Time lapse microscopy revealed that cell movement and reorganization were involved in spheroid formation. Ultrastructural evaluation using scanning and transmission electron microscopy indicated polarized cellular morphology and extensive cell-cell communication within spheroids. Bile canalicular structures were observed to surround each individual hepatocyte, forming an intricate three-dimensional continuous network of channels that appeared to end as pores/holes on the surface of the spheroid. The maintenance of differentiated cellular morphology coincided with preservation of hepatocyte viability and enhanced levels of tissue specific functions in spheroids. PMID- 8727044 TI - Tumorigenesis of rat mammary epithelial cells by N-nitroso-N-methylurea in an in vitro system: characterization of the microtumors. AB - Chemical carcinogenesis is a lengthy process that involves the rather loosely defined stages of initiation, promotion, and progression. Several model systems of mammary carcinogenesis have been designed to elucidate the mechanisms of chemical carcinogenesis. Most of these systems have included animal models. While organ specific chemical carcinogenesis can be initiated in these systems, the subsequent stages of promotion and progression are difficult to study in detail. Investigations on in vitro carcinogenesis have shown transformation of mammalian cells in culture; the transformational event, however, is difficult to discern within the monolayer culture. We have recently reported the development of an in vitro carcinogenesis system that allows both the initiation as well as the progression of mammary cells in a collagen gel matrix culture system. The cells transformed by a chemical carcinogen develop into discernible microtumors within the three dimensions of a collagen gel culture. Isolation of these microtumors from the collagen gel and subsequent culture in monolayer has produced cells capable of colony formation in soft agar. The present study further characterizes these microtumors originated in vitro by analysis of cell growth kinetics versus parallel control cells. In addition, flow cytometric and cytogenetic studies have been performed to investigate the chromosomal stability of these cells. It was also observed that the microtumors, produced in vitro from mammary epithelial cells of an inbred strain of rats, show the ability to form tumors upon transplantation into the fat pad of syngeneic hosts. PMID- 8727045 TI - Characterization of a primary hepatocyte culture system for toxicological studies. AB - An hepatocyte culture system was developed for potential use in toxicological studies in vitro. Rat hepatocytes were isolated by two-step collagenase perfusion and cultured on Vitrogen-coated Permanox dishes in a modified Chee's medium containing 1 microM dexamethasone and 1% dimethylsulfoxide. The cells remained highly viable for at least 10 d as determined by lactate dehydrogenase release and total protein levels. Albumin secretion into the medium, as a measure of differentiated function, was maintained at elevated levels over the course of 10 d in culture. A number of CYP activities were determined by the analysis of testosterone metabolism in freeze-thawed cells, diazepam metabolism in live cells, and specific assays for CYP 1A1/2, 2B1/2, 2E1, and 3A. Results of these assays indicated that a wide range of CYP isozymes were maintained, some activities were enhanced under the conditions of culture and some activities were inducible. Activities of the phase II enzymes, glutathione S-transferase and UDP glucuronosyltransferase, and glutathione levels were also maintained in the cultured hepatocytes for at least 6 d. These results strongly support the use of this hepatocyte culture system for in vitro toxicological studies. PMID- 8727046 TI - Effect of anoxia and reoxygenation on antioxidant enzyme activities in immortalized brain endothelial cells. AB - The effects of anoxia and reoxygenation on major antioxidant enzyme activities were investigated in vitro in immortalized rat brain endothelial cells (RBE4 cells). A sublethal anoxic period of 12 h was assessed for RBE4 cells using the neutral red uptake test. Anoxia markedly influenced the specific activity of catalase and superoxide dismutase, with no major effect on glutathione peroxidase or glutathione reductase. After 24 h postanoxia, the superoxide dismutase activity modulated by the presence or absence of oxygen returned to control value. Damage and recovery of RBE4 immortalized rat brain endothelial cells in culture after exposure to free radicals and other oxygen-derived species provides a useful in vitro model to study anoxia-reoxygenation trauma at the cellular level. PMID- 8727048 TI - Drug-induced reversion of progression phenotype is accompanied by reversion of AP 1 phenotype in JB6 cells. AB - Transformed JB6 cells can be stably reverted to nontransformed phenotype by AP-1 inhibiting gluccorticoid fluocinolone (FA) and cAMP elevator forskolin (FN), yielding stable revertants of promotion resistant (P-) and promotion sensitive (P+) phenotypes. AP-1 activity of nontransformed P- and P+ revertant clones was decreased under a variety of experimental conditions compared with their transformed counterparts. Moreover, AP-1 activity in P+ cells under anchorage independent conditions was induced by 12-0-tetradecanoyl-phorbol-13-acetate (TPA) while AP-1 activity in the reverted P- cells was not induced, just as observed for the original P+ and P- variants. Taken together these data suggest that changes in AP-1 activity may be one key mediator not only of forward progression but also of reversion of tumor cells to nontransformed phenotype. In addition, the higher transfection efficiency of the new reverted P- and P+ cells renders them useful for studying the role of transcription factors in tumor promotion. PMID- 8727047 TI - Cytokine production by a megakaryocytic cell line. AB - The regulation of megakaryopoeisis by cytokines is not yet well understood. It is possible that autocrine loops are established during megakaryocyte growth and differentiation, aiding in the maturation of these cells. The CHRF-288-11 human megakaryoblastic cell line has been examined for cytokine production in growing cells and cells stimulated to differentiate by the addition of phorbol esters. It has been demonstrated that these cells produce RNA corresponding to the interleukins IL-1 alpha, 1 beta, 3, 7, 8, and 11, granulocyte-macrophage colony stimulating factor (GM-CSF), stem cell factor (SCF), transforming growth factor beta (TGF-beta), tumor necrosis factor-alpha (TNF-alpha), interferon-alpha (INF alpha), and basic fibroblast growth factor (bFGF). Additionally, RNA corresponding to the receptors for IL-6, GM-CSF, SCF, INF-alpha, beta, bFGF, and monocyte colony stimulating factor (M-CSF) were also expressed by the cells. The receptor for TNF-alpha was detected immunologically. Analysis at the protein level demonstrated that significant amounts of INF-alpha, TNF-alpha, GM-CSF, SCF, IL-1 alpha, and a soluble form of the IL-6 receptor were produced by the cells. Addition of phorbol esters to CHRF-288-11 cells enhances their megakaryocytic phenotype; such treatment also results in increased secretion of INF-alpha, TNF alpha, and GM-CSF. These results suggest that potential autocrine loops are established during the differentiation of CHRF-288-11 cells, which may alter the capability of the cell to differentiate. These findings are similar to those recently obtained for marrow-derived megakaryocytes (Jiang et al.) suggesting that CHRF-288-11 cells provide a useful model system for the study of cytokine release during megakaryocyte differentiation. PMID- 8727050 TI - Comparative susceptibility of a canine cell line and bluetongue virus susceptible cell lines to a bluetongue virus isolate pathogenic for dogs. AB - Recently, bluetongue virus (BLU) serotype 11 was detected in diseased dogs that had been inoculated with live attenuated vaccine contaminated with this serotype of bluetongue virus (Akita et al., 1994). For various laboratory tests, BLU can be propagated in different cell cultures. No information was found in the literature about the possibility of propagating this virus in canine cells. To determine whether the BLU isolate from the contaminated canine vaccine (BLU-vac) is unique in its ability to replicate in canine cells, this virus was studied in parallel with U.S. prototype strains of BLU (serotypes 2, 10, 11, 13, and 17), in hamster lung (HmLu-1) and canine kidney (MDCK) cell cultures. In HmLu-1 cell cultures, the BLU-vac produced cytopathic effect (CPE) of the same type as the U.S. prototype BLU strains by 4 to 6 d postinoculation. In MDCK cell cultures, all of the BLU strains tested were able to replicate but did not produce CPE. The BLU-inoculated MDCK cells became persistently infected, and these cultures continued to produce infectious BLU even after six serial passages over 2 1/2 mo. In none of these cultures was CPE observed. In mixed cultures containing both HmLu-1 and MDCK cells, CPE first affected the HmLu-1 islands; subsequently, CPE spread also to the areas with MDCK cells. The silent persistent infection of the MDCK cells with BLU indicates that more stringent screening of the cells used in the production of live vaccines for various contaminating viruses is necessary. PMID- 8727049 TI - Effects of gamma-IFN and NGF on subpopulations in a human neuroblastoma cell line: flow cytometric and morphological analysis. AB - Neuroblastomas are neural crest-derived tumors that contain neuronal, melanocyte, and Schwann cell precursors. We examined the effects of treatment with gamma interferon (gamma-IFN) and nerve growth factor (NGF), alone, and in combination, on these progenitor subpopulations in the human neuroblastoma cell line, SH-SY5Y. Using fluorescence-activated flow cytometry (FACS), changes in expression of three differentiation-specific or -associated marker proteins, the 200 kD neurofilament protein, the myelin basic protein, and the S-100 protein, were analyzed. Growth rates and morphological changes associated with each treatment over the 2-wk incubation period were noted. The greatest effects were observed with combined IFN + NGF treatment. These were significant increases in expression of all three proteins, distinctive morphological signs of differentiation, and extensive inhibition of proliferation compared to control cultures. Treatment with NGF alone resulted in increased neurofilament protein expression and in the length and number of neurite extensions, but there was no effect on the growth rate. IFN induced striking morphological changes, significant inhibition of growth, and changes in protein expression that correlated with neuronal to non neuronal subpopulation shifts due to the death of differentiated cells. When treatment was discontinued after 15 d, the morphological changes induced by NGF were reversed within 2-3 d, while those induced by IFN +/- NGF were present up to 4 wk post-treatment. Small, neuroblastic colonies were observed throughout the treatment period and within 4-6 wk after the cessation of treatment this cell type fully reconstituted the cultures suggesting the presence of a stem cell. Our results indicate that treatment with gamma-IFN +/- NGF can regulate growth and induce, either stem cells or progenitor neuronal, Schwann and melanocyte subpopulations in the SH-SY5Y cell line to irreversibly differentiate. PMID- 8727051 TI - Accidental fatal drug overdoses in New York City: 1990-1992. AB - This study of all accidental fatal drug overdoses (N = 1,986) in New York City from 1990 to 1992, using medical examiner data, found that cocaine, often with opiates and ethanol, caused almost three-fourths of deaths, while opiates without cocaine caused roughly one-fourth of fatal overdoses. Only 5% of accidental drug fatalities were caused by drugs other than cocaine or opiates. This is a marked departure from the results of studies in the early 1980s when opiates prevailed as a cause of accidental fatal overdoses. In this study the highest cocaine overdose rates were found among males, African-Americans, and Latinos. Rates of opiate overdose without cocaine did not differ in regard to race/ethnicity except for low rates among Asians and other ethnic groups. There was a marked increase in the rate of combined cocaine and opiate overdoses from 1990 to 1992 and a more gradual but steady increase of overdoses due to opiates without cocaine during that time period. Overdoses due to drugs other than cocaine or opiates showed no increase during that time period. PMID- 8727052 TI - Familial factors in illicit drug abuse: an interdisciplinary perspective. AB - Research on familial factors in illicit drug abuse has been approached from the perspectives of psychiatric epidemiology, which aims to assess familial clustering of disorders, and clinical psychiatry, which focuses on intrafamilial dynamics. To date, however, these two research traditions have existed in relative isolation from each other, and there has been little consideration given to enhancing the understanding of familial factors in drug abuse by combining these approaches. The intention of this article is to critically review the literature in these two areas and to offer suggestions as to how the strengths from each could be applied in a comprehensive research approach. PMID- 8727053 TI - The significance of a coexisting opioid use disorder in cocaine dependence: an empirical study. AB - This study was undertaken to compare cocaine-dependent patients with and without an opioid use disorder, in an effort to identify important clinical similarities and differences between the two groups. Ninety patients hospitalized for cocaine dependence were divided according to whether or not they had a coexisting opioid use disorder; 32 patients (35.6%) had an opioid diagnosis and 58 (64.4%) did not. The groups were compared on substance use histories, psychopathology, severity of drug-related problems, and 3-months cocaine use outcome. Patients with an opioid diagnosis had more current major depression, antisocial personality disorder, and other substance use disorders. They also had higher medical and drug scores on the Addiction Severity Index, and longer cocaine use histories. Nonetheless, 3 month cocaine use outcome was similar for both groups. Interestingly, patients with an opioid diagnosis engaged in more treatment during the follow-up period. Cocaine-dependent patients with a coexisting opioid use disorder may thus constitute a subgroup with worse prognostic characteristics, for whom differential treatments may improve outcome. PMID- 8727054 TI - Treatment of perinatal cocaine addiction: use of the modified therapeutic community. AB - Treatment outcome was evaluated for perinatal cocaine addicts admitted to a hospital-based day treatment clinic, organized as a modified therapeutic community (TC) modality. The perinatal program consisted of a specialized track for women embedded within this larger coed day treatment clinic. A total of 87 perinatal women (28 pregnant at intake, the remaining 59 postpartum) were compared with cohorts of nonperinatal women (N = 63) and men (N = 158) admitted during the period of evaluation (September 1989 through December 1993). In logistic regression analysis, successful discharge urine status (last three urines prior to discharge drug-free) was associated with current child custody involvement (odds ratio = 2.80, 95% C.I. = 1.16-6.72), entering treatment when not postpartum (odds ratio = 0.15, 95% C.I. = 0.05-0.42), and taking psychiatric medication (odds ratio = 2.04, 95% C.I. = 1.11-3.72). Both pregnant and postpartum perinatal women showed a similar pattern of shorter treatment as compared with nonperinatal women and male clients, averaging 2 months of treatment as compared with 4 months for nonperinatal clients. Factoring out pregnancy and postpartum status, the women in treatment fared as well as men with respect to both retention and discharge urine standings. This finding indicates that programmatic modifications need to address specific perinatal issues and not gender issues per se. Also, a differential pattern in discharge urine status of women who enter treatment while pregnant vs. those who enter when postpartum suggests that outreach and recruitment be targeted, but not limited, to pregnant women. PMID- 8727055 TI - The response of injection drug users to free treatment on demand: implications for HIV control. AB - Injection drug use is a major risk factor for human immunodeficiency virus (HIV) infection and drug treatment is widely recognized as a core component of the public health effort to limit the spread of HIV. The assumption is frequently made that lack of immediate access to treatment is a significant barrier to the success of this effort. However, little empirical data exist to support this belief. We conducted a trial of no-cost outpatient drug-free treatment made available on demand to a cohort of out-of-treatment injection drug users (IDUs) in Portland, Oregon, through a coupon program. Of 824 IDUs, 272 (33%) expressed an interest in treatment, 225 (27%) accepted a coupon, 66 (8%) redeemed a coupon, and 9 (1%) remained in treatment for 6 months. These numbers indicate that simply enhancing access is not adequate. Additional strategies to increase motivation to enter and remain in treatment are needed if drug treatment is to play an important role in reducing the spread of HIV among injection drug users, their sexual partners, and their infants. PMID- 8727056 TI - Alcohol and drug abuse in patients with physical disabilities. AB - Alcohol and other drugs of abuse (AODA) are of great medical and social concern. Patients with physical injury requiring rehabilitation may be at particular risk of AODA due to pain, physical handicaps, mood disturbances, vocational difficulties, and problems of self-image. Their access to AODA, however, is often temporarily or permanently limited. In this literature review, we have explored various aspects of AODA in physically disabled patients. Frequently, AODA are involved in the cause of physical injuries. The average use and abuse of alcohol prior to injury was high. Postinjury alcohol use and abuse frequently declined or remained unchanged. Some non-alcohol drug use and problems increased postinjury, particularly during initial periods. Postinjury abuse of AODA is particularly important with regard to the disruption of the rehabilitation process. Use of adequate control groups and analysis at multiple postinjury time points is recommended in future investigations. PMID- 8727057 TI - A comparison of the Johnson Intervention with four other methods of referral to outpatient treatment. AB - The Johnson Intervention is a therapeutic technique in which members of the person's social network confront him or her about the damage the drinking or drug use has caused and the action they will take if treatment is refused. It is highly effective in engaging and retaining clients in inpatient treatment, but, since initial evaluations, two trends have emerged in the field. First, there has been an increase in the use of outpatient treatment and, second, a number of variations of the Johnson Intervention have been developed. The purpose of this study was to compare the effectiveness of the Johnson Intervention with four other methods of referral to outpatient treatment. The other methods included two naturally occurring types of referral (coerced and non-coerced) and two less intense and less costly variations of the Intervention (Unrehearsed and Unsupervised). Effectiveness was determined by both entry into and completion of treatment. A retrospective study was conducted on a sample of 331 cases drawn from an alcohol and other drug treatment agency. Those who had undergone the Johnson Intervention were more likely to enter treatment than those in any of the four other groups. Of those that entered treatment, the Johnson Intervention and the coerced referral groups were equally likely to complete treatment, and both groups were more likely to complete treatment than those in the other three types of referral. Although the Johnson Intervention was the most effective, the variations did show some measure of success and can be viewed as part of an Intervention continuum. PMID- 8727058 TI - Relationship of hardiness to alcohol and drug use in adolescents. PMID- 8727059 TI - Pathways to self-help among women for sobriety. AB - Women for Sobriety represents a self-help option oriented toward positive thinking and behavior modification. Formed in 1975, there are now approximately 125 active WFS groups holding weekly meetings in the United States and Canada. Using data collected from a 1991 membership survey (n = 600, response rate = 73%), this paper chronicles WFS members' pathways to recovery. We describe turning points in seeking help, things women tried at first to contain their drinking, exposure to other treatment approaches, and referrals to WFS from formal treatment programs. Only 15% of the respondents sought treatment because they had been confronted about their drinking, while physical symptoms or emotional problems represented the turning point for over half the women. Another fifth said their life had gotten out of control, and these were the ones to achieve sobriety most quickly. Almost all WFS members had sought professional help for their drinking, and three-fourths have undergone individual therapy suggesting that WFS members are fairly open to a psychological approach such as WFS offers. WFS philosophy is based on the belief that behavior is predicated on thoughts, so the program teaches members that maintaining sobriety must involve the realization that negative emotions are destructive-and that members can control how much they allow problems to bother them. Goals of membership include abstinence, improved self-esteem, and spiritual and emotional growth. About a third of the respondents currently attend AA, and no differences in length of sobriety associated with current AA attendance were found within the WFS membership. However, women who sought professional help in response to the turning point experience but who also attended AA that year were most likely to have achieved sobriety within a year of the turning point. PMID- 8727060 TI - Characteristics of participants in secular organizations for sobriety (SOS). AB - Self-help groups have assisted many in their recovery from alcohol use disorders. Although Alcoholics Anonymous (AA) is the largest self-help organization for addressing alcohol problems, no single organization can be suited to the needs of all alcohol abusers. For example, some alcoholics have chosen not to affiliate with AA because of objections to religious or spiritual references in the 12 steps of AA. It was largely for this reason that a secular self-help organization Secular Organizations for Sobriety (SOS)-was established. This article provides an overview of SOS. Also described are the results of a survey of SOS participants. These data provide a preliminary picture of SOS members and will, we hope, stimulate further research on this organization and its effectiveness. PMID- 8727061 TI - Cat scratch disease and acquired immunodeficiency disease: diagnosis by transmission electron microscopy. AB - A 33-year-old, homosexual, cat-owning, African-American man with human immunodeficiency virus infection by positive serologic tests and acquired immunodeficiency syndrome by CD4 lymphocyte count alone (39 cells/mL) presented with a one-year history of intermittent fever, weight loss, and generalized lymphadenopathy. A malignant lymphoma was suspected clinically. Light microscopic study of a left inguinal lymph node biopsy specimen revealed effacement of the lymph node architecture by a diffuse infiltrate of large, atypical reticulum cells, loose, patchy granulomatous inflammation, diffuse hyaline fibrosis, diffusely proliferated blood vessels, and multifocal degeneration and necrosis. Lymph follicles were absent and lymphocytes were moderately depleted. Microorganisms were not seen in lymph node sections stained with special histochemical stains (including the Warthin-Starry stain). These light microscopic changes were considered suggestive of a malignant lymphoma, especially Hodgkin's disease. The diagnosis of cat scratch disease (CSD) became apparent only after transmission electron microscopic study of the lymph node revealed clusters of small, pleomorphic bacteria in degenerated collagenous tissue and in blood vessel walls. This case illustrates the value of transmission electron microscopy in making the diagnosis of CSD, especially when light microscopic changes are superimposed on those of late human immunodeficiency virus infection of the lymph node. PMID- 8727062 TI - Bacteria in biopsies of human hypochlorhydric stomach: a scanning electron microscopy study. AB - A long-lasting condition of hypochloridria leads to a bacterial growth both in the gastric lumen and biopsies of human stomach. Some of these bacteria are probably involved in gastric carcinogenesis, due to their capacity of nitrosation. This study was carried out on biopsies taken during endoscopy from both gastric antrum and the body of patients with or without hypochloridria. Scanning electron microscopy observation shows that bacteria, other than Helicobacter pylori, found in hypochloridria, can be located not only over but also into and under the mucus layer covering the gastric epithelium. In such areas, mechanical and biochemical damage may occur. PMID- 8727063 TI - The human blood-testis barrier in impaired spermatogenesis. AB - The aim of this study was to explore the competence of the blood-testis barrier (BTB) using electron opaque tracers in diverse human testicular pathologies associated with Sertoli cell only syndrome. Two groups of patients were studied: (1) those with complete depletion (absence) of germ cells, and (2) those with severe germ cell depletion but with some germ cells left in the seminiferous epithelium. The first situation was associated with cryptorchidism with absence of germinal cells, idiopathic cases of aplasia of germ cells, peritumoral areas surrounding small seminomas where the seminiferous tubules were observed to contain a predominant population of Sertoli cells, or long estrogen treatment. The second was found also in cryptorchidism with early germ cells, idiopathic azoospermia, and oligospermia associated with sterility. In the first situation, seminiferous tubules lacked lumen and Sertoli cells had immature morphological characteristics, i.e., oval nuclei with smooth profiles, even heterochromatin distribution and a single, small nucleolus. Inter-Sertoli tight junctions were tortuous, interrupted, and mostly perpendicular to the basal lamina. Lanthanum hydroxide or nickel nitrate permeated most of the inter-Sertoli spaces, indicating disruption of the BTB. In the second situation, seminiferous tubules had a lumen, and Sertoli cells exhibited a mature appearance with large tripartite nucleoli and irregular, highly infolded nucle-olemma. Only spermatogonia or primary spermatocytes showing diverse degrees of cell involution were found. Numerous inter-Sertoli tight junctions, uninterrupted and parallel to the basal lamina, stopped the electron opaque intercellular tracers close to it; this meant the assembly of a competent BTB. Therefore, a close correlation was found between morphological parameters of Sertoli cell maturity, including their tight junction organization, and BTB integrity. PMID- 8727064 TI - Globular smooth-tubule aggregates in thanatophoric dwarfs. AB - A presumably new structure that we have called "globular smooth-tubule aggregate" was found in the cytoplasm of chondrocytes of 4 thanatophoric dwarfs. To the best of our knowledge these sharply delineated, rounded, sieve-like bodies of distinctive morphology have not been reported to occur in any other site or situation. Hence these structures may be of some diagnostic value. Continuity between the rough endoplasmic reticulum and smooth tubules in the aggregates was detected. Therefore it is suggested that smooth tubules in these aggregates derive by growth and extension from the rough endoplasmic reticulum. PMID- 8727065 TI - Cryofixation, cryosubstitution, and immunoelectron microscopy: potential role in diagnostic pathology. AB - In immunoelectron microscopic investigations, retention of antigenic sites is crucial. Methods for preparing samples for conventional electron microscopy involve chemical fixatives followed by dehydration in organic solvents and embedding in plastic resins, all procedures potentially detrimental to antigenicity. Cryomethods provide a physical fixation alternative for the preparation of biological samples for ultrastructural, immunocytochemical, and microanalysis studies without the use of any chemicals. This can be particularly useful in diagnostic pathology, providing an alternative to conventional fixation methods which sometimes destroy the antigen in question. The recent development of a portable cryofixation device, the PS1000 Portable Metal Mirror Ultra-Rapid Cryofixation Unit (Delaware Diamond Knives, Inc., Wilmington, DE, USA), provides an opportunity to freeze tissue immediately after procurement for use in diagnostic immunocytochemistry studies. This feasibility study examined the quality of tissue preservation with this device, in terms of both preservation of cellular ultrastructure and immunolabeling. Human tonsil and thymus tissue was slam frozen and, after cryosubstitution in Lowicryl K11M, was examined by immunoelectron microscopy. Good ultrastructural preservation was obtained and reasonable immunolabeling with antibodies to AE1/AE3 keratin filaments was also observed. PMID- 8727066 TI - Adult-type granulosa cell tumor, presenting as extraovarian metastases: ultrastructural diagnosis. AB - All granulosa cell tumors have the potential to recur locally or metastasize, in spite of apparent complete surgical excision. Long-term follow-up is particularly important in these tumors, since recurrences may occur many years after the initial diagnosis. This report highlights the ultrastructural findings that led to the diagnosis of metastatic granulosa cell tumor in two patients presenting with extraovarian masses. In patient 1 the information about an ovarian granulosa cell tumor removed 9 years before the current admission was obtained only after the metastatic tumor was correctly diagnosed. In patient 2 the ovarian primary was found after abdominal wall involvement by granulosa cell tumor was diagnosed. Thus, in both cases the clinical circumstances were atypical and electron microscopic examination of the tumors was essential to make the diagnosis of granulosa cell tumor with confidence. Based on the features observed in these two extraovarian tumors as well as eight other primary ovarian granulosa cell tumors from our files, a consistent ultrastructural profile is identified that can be very useful for diagnosis. PMID- 8727067 TI - Observation of psammoma bodies in cultured meningiomas: analysis of three dimensional structure using scanning and transmission electron microscopy. AB - In contrast to the inner structure, three-dimensional structure of psammona bodies in meningiomas is not well defined. This study examined three cultured meningiomas, in which surface observation of psammoma bodies might be easier than in the tumor tissues since influence of interposing connective tissue is minimized in tissue culture. Early culture revealed that psammoma bodies with frank calcification were suspended in the tissue culture medium, and so were they collected, centrifuged, and then processed for electron microscopy. Ultrastructurally, psammoma bodies were mostly spherical in shape and composed of a core of dense calcification and surrounding collagen fiber bundles. Apart from psammoma bodies, round bodies with concentric lamination like a transversely cut onion were frequently noted by light microscopy. These bodies were composed mainly of tangles of collagen fibers emerged from surrounding tumor cell processes. The results suggest that psammoma bodies in meningiomas arise in part from meningothelial whorls due to collagen production by tumor cells followed by obliteration and disappearance of tumor cell processes, although some of the alternative pathways for psammoma body formation proposed by other investigators cannot be ruled out by this study. PMID- 8727068 TI - Morphologic changes in intercostal muscle tissue associated with a viper (Agkistrodon halys blomhoffi) bite. AB - This study examined a 73-year-old woman who had been bitten in the right thumb by a viper (Agkistrodon halys blomhoffi). She suffered acute renal failure and respiratory failure after the bite. On the 20th day, respiratory failure persisted, although she had recovered from renal failure. Intercostal muscles were obtained by biopsy on the 40th day after the bite. Specimens were observed with enzyme-histochemical and electron microscopic techniques. The size of muscle fibers varied. Most of the smaller muscle fibers were enzyme-histochemically type 2C fibers. This was considered to be the regenerative stage of acute rhabdomyolysis. Ultrastructurally, two opposite types of changes, degeneration and regeneration, were mixed in the same areas. Muscle fibers might be continuously degenerated, and the regenerative fibers might also be degenerated. This study suggests that some slow allergic reactions to snake venom might bear some relation to the degenerative changes. PMID- 8727069 TI - Ultrastructural heterogeneity of acquired intradermal melanocytic nevus cells. AB - The present ultrastructural evaluation of 12 acquired intradermal melanocytic nevi revealed that in contrast to the nested epithelioid melanocytic nevus cells of the upper dermis, the spindle nevus cells of the deep dermis showed perineurial differentiation, exhibiting a spindly configuration characterized by a melanosome-free cytoplasm that showed extremely slender bipolar contour and contained abundant intermediate filaments, a decreased number of cytoplasmic organelles, and, significantly, a fair number of plasmalemmal pinocytotic vesicles. The nevic corpuscles were found to consist of laminated slender cytoplasm showing subcellular conformation similar to that of the spindle nevus cells. By immunohistochemistry, many spindle nevus cells and nevic corpuscles were immunoreactive for nerve growth factor receptor. All the nevus cells were immunoreactive for vimentin and S-100 protein, and negative for protein gene product 9.5, epithelial membrane antigen, Leu-7, and myelin basic protein. Characteristically, protein gene product 9.5 immunohistochemistry revealed numerous immunoreactive axons intermingled with the spindle nevus cells in the deep portion. All the PGP9.5-immunoreactive axons were observed by immunoelectron microscope to be unmyelinated and always ensheathed by a thin cytoplasmic process of Schwann cells but not nevus cells. These findings indicate that differentiation plasticity exists in the various nevus cells, with the epithelioid nevus cells and the spindle nevus cells displaying more ultrastructural and immunophenotypical characteristics of melanocyte and perineurial cells, respectively, suggesting that a pluripotential cell of neural crest origin accounts for the histogenesis of this lesion. PMID- 8727070 TI - Ultrastructural investigation of lead-induced intranuclear inclusion bodies in mice. AB - By means of optical and electron microscopy and by atomic absorption spectrophotometry in a graphite chamber, this study evaluated the effect of temperature (22-35 degrees C) on lesions in the kidney, liver, and brain, and on concentrations of lead caused by the administration of 2 and 5 mg/kg/IP of lead acetate to Swiss mice. The most pronounced effects were observed in the kidney and in groups of animals receiving the highest doses (5 mg/kg at 22 and 35 degrees C). These effects consisted of significantly higher (p < .05) lead concentrations in the tissues, a significant decrease (p < .05) in kidney weights, and progressive kidney atrophy and fibrosis with, at the ultrastructural level, constant intranuclear inclusions, which were also observed in the cytoplasm of renal and endothelial cells. PMID- 8727071 TI - Subchronic toxicity of 2,4,4'-trichlorobiphenyl in the rat liver: an ultrastructural and biochemical study. AB - 2,4,4'-Trichlorobiphenyl or PCB congener 28 was given to Sprague-Dawley weanling rats and the experimental diets were prepared by mixing the congener in 4% corn oil. The congener was administered to animals placed in four groups, each comprising 10 males or females. The diets contained 0.05, 0.5, 5, or 50 ppm congener. The fifth or control group comprised animals that received diets mixed with corn oil. Thirteen weeks after commencement of dosing, animals were euthanized and liver specimens were harvested from the animals and prepared for electron microscopy and biochemical analyses. The hepatocyte architectural modifications included an augmentation of SER profiles and an elevation of peroxisome numbers in animals regardless of gender, and mitochondrial abnormalities in the females only. Mitochondrial aberrations consisted of abnormal shapes and cristae in atypical orientation. The alterations were revealed in animals of the 5-and 50-ppm groups and were more extensive in the females. Ethoxyresorufin-O-de-ethylase activity was significantly high in the animals of the 50-ppm group. The results suggest that the female rats were more sensitive than the males to congener 28, and the no observable adverse effect level (NOAEL) was believed to be 0.5 ppm for the congener. PMID- 8727072 TI - Case for the panel. Unusual ultrastructural association between erythrocytes and glomerular endothelial cells in patients with membranoproliferative glomerulonephritis. PMID- 8727073 TI - Interferons in precancer and cancer prevention: where are we? AB - Clinical research on interferons (IFN) still focuses on the treatment of advanced cancer. The research strategy eventually must be reevaluated. The cellular minimal deviations that are seen in early phases of carcinogenesis might be the most rational target for immune interventions. That biologic response modifiers have considerable capacity to prevent induction and development of malignant neoplasias has been demonstrated in several animal systems. Even the few clinical studies available at present on the treatment of preneoplastic lesions with IFN have definitely shown more success than those involving treatment of advanced tumors. In addition, there is experimental evidence that IFN might be suitable candidates for immunoprevention. The major problems hampering a wider application of IFN in immunoprevention is that they cause adverse effects. Unfortunately, we do not know much about the specific mechanisms involved in the immune control of human tumor development during the initial and the latency phases of carcinogenesis. More research is needed in this area. In this article the state of the art of using IFN for treating preneoplastic lesions is reviewed, and also we report some of our experimental results on IFN and anticancerogenesis. PMID- 8727074 TI - Effects of interferon-alpha, beta, and gamma on the function of differentiated leukemic HL-60 cells induced by 1,25-dihydroxyvitamin D3. AB - The differentiation of HL-60, a human leukemic cell line, into monocyte-like cells (D3-HL-60 cells) is induced by 1,25-dihydroxyvitamin D3 (D3). We examined the effects of interferon (IFN) treatment of D3-HL-60 cells on the expression of cell surface antigens, the phagocytic activity for fluorescent beads, production of oxygen radicals, and intracellular growth of Legionella pneumophila. Activation of D3-HL-60 cells with IFN-gamma, Beta, and alpha for 24 h significantly increased expression of CD16, CD36, CD71, and HLA-DR antigens. IFN gamma markedly enhanced the phagocytic activity of beads in D3-HL-60 cells. There was no significant difference in phagocytic activity between cells exposed to IFN alpha or beta and untreated D3-HL-60 cells. IFN-alpha, beta, and gamma enhanced production of oxygen radicals, including superoxide, by D3-HL-60 cells. Superoxide production was enhanced to the greatest degree by IFN-gamma, followed by IFN-beta and then IFN-gamma. Intracellular growth of L. pneumophila in D3-HL 60 cells was inhibited by interferons (IFN-gamma > beta > gamma). Similar results were obtained in human mononuclear cells. These data indicate that interferons can act as biologic response modifiers not only in human mononuclear cells but also in differentiated leukemic cells. Our results may have implications for the development of differentiation therapy for treatment of leukemia. PMID- 8727077 TI - Partial inhibition of vesicular stomatitis virus by the interferon-induced human 9-27 protein. AB - To determine whether the interferon-induced 9-27 protein of human cells contributes to the antiviral state, we expressed the 9-27 cDNA under the control of a constitutive promoter and assayed transfected cells for enhanced virus resistance. The intracellular distribution of 9-27 resembled that of cytoskeleton associated proteins. Analysis at the single-cell level by indirect immunofluorescence revealed that mouse cells expressing 9-27 were less permissive for vesicular stomatitis virus than control cells not expressing 9-27. No significant inhibition of influenza virus was observed. When tested in parallel, 9-27 was found to have less powerful antiviral activity toward vesicular stomatitis virus than the interferon-induced MxA protein. Thus, 9-27 joins the family of interferon-induced proteins with intrinsic antiviral activity. PMID- 8727075 TI - Effect of therapy with recombinant human interferon-gamma on the release of nitric oxide by neutrophils and mononuclear cells from patients with chronic granulomatous disease. AB - The aim of this study was to investigate the effect of recombinant human interferon-gamma (rHuIFN-gamma) therapy on the release of nitric oxide (NO) by neutrophils (NEU) and mononuclear cells (MON) from patients with chronic granulomatous disease (CGD). Five patients with this rare disease received rHuIFN gamma (50 micrograms/m2 of body surface, given by subcutaneous injection three times a week) for 6 months. Clinical and laboratory evaluations were performed before and after 1 and 6 months of rHuIFN-gamma therapy. Nitric oxide release by NEU and MON was assessed by the ability of these cells to inhibit thrombin induced washed platelet aggregation. The nitrite (NO2-) and nitrate (NO3-) levels in the supernatant of cultured NEU and MON, as well as in plasma and urine (24 h diuresis), were quantified by high-performance liquid chromatography (HPLC). Conventional immunologic tests for assessing phagocyte and lymphocyte functions and humoral immunity were also performed. Therapy with rHuIFN-gamma for 6 months did not enhance NO synthesis by NEU or MON from the patients with CGD. The urinary but not plasma levels of NO2- and NO3- were elevated after rHuIFN-gamma therapy. Phagocyte and lymphocyte functions as well as humoral immunity were not affected by rHuIFN-gamma therapy. Although few patients were available for the study, we conclude that therapy with rHuIFN-gamma for 6 months did not enhance the synthesis of NO by NEU and MON in CGD patients. Whether the increased excretion of NO2- and NO3- in the urine of CGD patients after rHUIFN-gamma therapy reflects an induction of NO-synthase in cells other than leukocytes remains to be investigated. PMID- 8727076 TI - Potential role of interleukin-1 in the development of bronchopulmonary dysplasia. AB - Increased activities of inflammatory mediators unopposed by their inhibitors contribute to chronic lung injury and impaired healing in BPD. The deleterious effects of IL-1 beta, a cytokine involved in inflammation and host defense, are blocked by IL-1 receptor antagonist (IL-1Ra). We proposed that an imbalance of IL 1 beta and its inhibitors may contribute to the development of BPD. To determine the relative antigen concentrations of IL-1 beta and IL-1Ra and functional IL-1 activity in lung lavage of infants at risk for BPD, lung lavage was serially obtained from 1 to 28 days from 17 infants with evolving BPD, 13 infants with self-limited RDS, and 6 controls ventilated for nonpulmonary reasons. Overall, there was a high correlation between IL-1 beta antigen concentration and IL-1 activity (r = 0.82, p = 0.0001). There were no significant differences among the groups for lung lavage variables on day 1. However, in infants who developed BPD, IL-1 beta antigen concentration and IL-1 activity increased 16- and 61-fold, respectively, during the first week. IL-1Ra remained relatively unchanged during the first month. IL-1 beta/IL-1Ra antigen ratio was significantly higher on days 5 (median 0.024) and 7 (median 0.025) compared with day 1 (median 0.004), p < 0.05. These results suggest that a relative imbalance of IL-1 beta and IL-1Ra may contribute to prolonged inflammation in BPD. PMID- 8727078 TI - Deficient tumor necrosis factor secretion by cord blood mononuclear cells upon in vitro stimulation with Listeria monocytogenes. AB - We examined the production of tumor necrosis factor (TNF) by mononuclear cells (MNC) after incubating adult or cord blood MNC with Listeria monocytogenes in vitro. With adult MNC cultures, we found that TNF activity reached a peak at 6 h (606 +/- 120 x 10(3) units/liter) and declined to the baseline by day 3. In contrast, using cord blood MNC, we found that TNF activity increased gradually reaching a peak at 24 h. In addition, the peak TNF activity using newborn MNC (189 +/- 26 x 10(3) U/liter) at 24 h was still lower than the peak using adult MNC at 6 h (p < 0.0002). In seeking an explanation for the decreased TNF secretion from newborn MNC, we examined the possibility that newborn cells produce TNF but failed to secrete it. However, lysates of newborn cells contained functionally and antigenically less TNF than adult cells. Based on these observations, we conclude that the overall TNF production by newborn cells incubated with L monocytogenes is decreased compared with similarly stimulated adult cells. PMID- 8727079 TI - Indirect stimulatory effects of murine interleukin-12 on in vitro production of nitric oxide by mouse peritoneal cells. AB - The effect of murine interleukin-12 (IL-12) on L-arginine-dependent biosynthesis of nitric oxide (NO) by mouse peritoneal cells was evaluated. Interleukin-12 was found to trigger considerably enhanced production of NO in a dose-dependent manner. Antibody neutralization studies indicated that the effect of IL-12 was mediated by IFN-gamma without apparent participation of TNF-alpha. Synergistic effects of IL-12 plus lipopolysaccharide (LPS) were also observed. Our data thus provide evidence that IL-12 is a powerful but indirect modulator of NO formation. These findings may contribute to the better understanding of various biologic effects of IL-12. PMID- 8727080 TI - Induction of tumor necrosis factor alpha expression in human T lymphocytes following ionizing gamma irradiation. AB - In this work, we present evidence that enriched human peripheral blood T lymphocytes, depleted of contaminating monocytes, rapidly express tumor necrosis factor alpha (TNF-alpha) mRNA when exposed to low doses of gamma-irradiation. In total PBL, TNF-alpha mRNA accumulation increased threefold as early as 30 minutes following exposure to 4 Gy and then declined to the baseline level by 3-5 h, as measured by the reverse transcriptase-polymerase chain reaction (RT-PCR). The increase in TNF-alpha mRNA was also observed in populations of enriched T cells and decreased when the dose of irradiation was increased to 10 Gy, strongly suggesting that T lymphocytes, the most radiosensitive cells of the body, contributed directly to the increase of TNF-alpha mRNA. A good correlation was found between mRNA expression and TNF-alpha protein secretion. Interestingly, a eightfold increase in glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA accumulation was also detected in both PBL and enriched T cells irradiated at 4 Gy for 3 h compared with unirradiated cells. This irradiation effect was almost completely abolished, however, following exposure to 10 Gy. Together these data suggest that T cells are responsible for the irradiation-induced expression of TNF-alpha and GAPDH. PMID- 8727081 TI - Tumor necrosis factor effects on ascites formation in an experimental tumor model. AB - In humans, treatment of malignant ascites with bolus TNF leads to resolution of the ascites. In an experimental model NMRI nude mice were inoculated intraperitoneally with human NIH-OVCAR3 adenocarcinoma cells, resulting in production of ascites and intraperitoneal tumor growth. Ascites formation and tumor growth after IP injection of recombinant human TNF was determined. Depending on the treatment schedule, a dual effect of TNF on the development of ascites was seen. Doses of TNF (1-10 micrograms/g) given once per week completely prevented ascites production, whereas the same doses of TNF given on a daily schedule induced enhanced ascites formation in an inverse TNF dose relationship. The area of tumor cell-covered peritoneal lining corresponded to these findings, indicating a correlation of tumor mass with ascites production. In an attempt to prevent renewal of ascites after drainage, neither inhibition nor enhancement in ascites production was seen when TNF was given five times per week. However, doses of 10 micrograms/g of TNF once per week led to almost complete inhibition of ascites reappearance. Histological examination of animals that received repeated TNF treatment demonstrated chronic peritonitis with strong stromal proliferation, angiogenesis, and increased adhesion of tumor cells to the peritoneum. PMID- 8727082 TI - Effect of space flight on cytokine production and other immunologic parameters of rhesus monkeys. AB - During a recent flight of a Russian satellite (Cosmos #2229), initial experiments examining the effects of space flight on immunologic responses of rhesus monkeys were performed to gain insight into the effect of space flight on resistance to infection. Experiments were performed on tissue samples taken from the monkeys before and immediately after flight. Additional samples were obtained approximately 1 month after flight for a postflight restraint study. Two types of experiments were carried out throughout this study. The first experiment determined the ability of leukocytes to produce interleukin-1 and to express interleukin-2 receptors. The second experiment examined the responsiveness of rhesus bone marrow cells to recombinant human granulocyte-macrophage colony stimulating factor (GM-CSF). Human reagents that cross-reacted with monkey tissue were utilized for the bulk of the studies. Results from both studies indicated that there were changes in immunologic function attributable to space flight. Interleukin-1 production and the expression of interleukin-2 receptors was decreased after space flight. Bone marrow cells from flight monkeys showed a significant decrease in their response to GM-CSF compared with the response of bone marrow cells from nonflight control monkeys. These results suggest that the rhesus monkey may be a useful surrogate for humans in future studies that examine the effect of space flight on immune response, particularly when conditions do not readily permit human study. PMID- 8727083 TI - Our altruistic mission. PMID- 8727084 TI - Pyogenic infectious spondylitis in children. The evolution to current thought. AB - The historic evolution of pyogenic infectious spondylitis from its initial descriptions to its current characterization illustrates the degree to which our means of perception and evaluation influence our understanding of an illness. As new concepts resulting from medical innovations challenge traditional beliefs regarding the etiology and optimum treatment of pyogenic infectious spondylitis in children (commonly termed discitis), controversies have arisen. This review attempts to clarify these issues by reconstructing their historic basis and delineating the limitations of our current knowledge. In our opinion, this exercise demonstrates the similarities between pyogenic infectious spondylitis in children and adults and provides support for the management of this illness at all ages with parenteral antibiotics, cast or brace immobilization when needed for symptomatic relief, and, rarely, operative debridement. PMID- 8727085 TI - Perioperative complications in patients with sickle cell disease. An orthopedic perspective. AB - Osteonecrosis of the femoral head has been reported to occur in 19% to 31% of patients with sickle cell disease, with the condition often being bilateral. Current surgical options for these patients include various forms of arthrodesis, resection arthroplasty, osteotomy, and uncemented hip arthroplasty. Poor surgical outcome, coupled with frequent perioperative medical complications, makes the treatment of these patients very challenging. A case report of a 33-year-old black woman with a 21-year history of sickle cell disease who underwent hip arthroplasty and developed sickle chest syndrome is presented. PMID- 8727086 TI - The prevention of complications with the Gamma Locking Nail. AB - The general acceptance of the Gamma Locking Nail (Howmedica, Rutherford, New Jersey) in the United States has been slow, because of reported intraoperative fractures of the shaft of the femur occurring with use of this nail. A series of 63 consecutive intertrochanteric and subtrochanteric fractures of the femur is reported. There were no intraoperative or postoperative fractures of the shaft of the femur in this series. Intraoperative fractures are avoided by appropriate over-reaming of the shaft of the femur, insertion of the device by hand only, and drilling no more than one hole for each distal locking screw. The use of a hammer during insertion of the Gamma Nail will be associated with femoral shaft fractures. Proper positioning of the lag screw in the head and neck of the femur will prevent migration of the lag screw in the head of the femur. The Gamma Nail has a definite place in the treatment of fractures of the proximal femur. PMID- 8727087 TI - Complete endoscopic carpal canal decompression. AB - We made a model of the endoscopic decompression of the carpal canal in clinical cases. The model entailed the release of the transverse carpal ligament, ie, the flexor retinaculum, first; then the transverse fibers: deep layer of the midpalmar fascia or distal portion of the flexor retinaculum; and, finally, release of the forearm fascia. Carpal canal pressure was measured using the continuous infusion technique, and the carpal canal was observed endoscopically at each step. Carpal canal pressure data were analyzed by using the Wilcoxon matched pairs signed-rank test. When the transverse carpal ligament and the transverse fibers were divided, carpal canal pressure was significantly statistically lower than that with release of the transverse carpal ligament alone. We conclude that release of both the transverse carpal ligament and the transverse fibers are essential for complete decompression of the carpal canal in endoscopic surgery. PMID- 8727088 TI - Bilateral spontaneous patellar tendon rupture. AB - This report describes a bilateral spontaneous patellar tendon rupture in a patient without pre-existing systemic disease. A possible association is recognized that may predispose a patient to bilateral spontaneous patellar tendon rupture. PMID- 8727089 TI - An unusual presentation of telangiectatic osteosarcoma. AB - Telangiectatic osteosarcoma (TOS) is a rare subtype of osteosarcoma that accounts for approximately 2% of all osteosarcomas. Presented is a case of a sacral mass that was originally thought to be a chordoma and was treated surgically but on pathologic examination was diagnosed as a TOS. A review of the literature on other sacral lesions is provided. The intention for presenting this case is twofold: to make physicians aware of the differential diagnoses of sacral lesions and to report an unusual presentation of this rare tumor. PMID- 8727090 TI - External rotation views in the diagnosis of posterior colliculus fracture of the medial malleolus. AB - A 25-year-old white man sustained a fracture dislocation of the right ankle with a displaced fracture of the lateral malleolus while playing basketball. He also sustained a fracture of the medial aspect of the distal tibia, in addition to an apparent deltoid ligament injury. The patient underwent open reduction and internal fixation of the lateral malleolus and insertion of a syndesmotic screw. Intraoperatively, external rotation radiographs of the ankle were obtained to determine the origin of the fracture of the medial side of the ankle joint. The radiographs clearly showed a minimally displaced fracture of the posterior colliculus. Posterior colliculus fracture can be occult. This fracture may not be clearly visible on plain antero-posterior or mortise-view radiographs and external rotation is necessary for its diagnosis. This fracture is usually minimally displaced and can be satisfactorily treated conservatively. The patient's leg was placed in a short-leg cast for 6 weeks. Follow-up radiographs revealed healing of the fracture. PMID- 8727091 TI - Changes occurring in health care and orthopedics. PMID- 8727092 TI - Staphylococcus infection complicating the application of a supracondylar nail to a fracture proximal to a total knee arthroplasty. PMID- 8727093 TI - Update on informed consent for stored tissue research. PMID- 8727094 TI - Hepatitis C virus reinfection in orthotopic liver transplant patients with or without concomitant hepatitis B infection. AB - Although recurrence of hepatitis C virus (HCV) in orthotopic liver transplant (OLT) patients is frequent, the relationship between HCV recurrence and graft pathology, particularly in patients who also have a history of hepatitis B virus (HBV), is unclear. The recurrence of HCV after OLT was determined by reverse transcriptase-nested polymerase chain reaction (RT-PCR) in the sera and livers of 41 patients with OLT, 32 of whom underwent transplants for HCV or HBV-related disease. Results were compared with liver function tests, liver histology (including HBV immunohistochemistry), and antibody status. HCV PCR was more frequently positive in OLT patients with a history of HCV only (59%) than in those with a history of both HCV and HBV (41%) or no history of viral infection (2%). Recurrent HCV (60% overall) was associated with mild elevation of liver function tests and mild to moderate hepatitis. In patients who underwent transplants for both HCV and HBV disease, hepatitis on biopsy was more frequently associated with recurrent HBV than with recurrent HCV. We conclude that graft reinfection with HCV, which is frequent in OLT patients with or without HBV recurrence, is usually associated with only mild to moderate hepatitic changes compatible with graft survival. PMID- 8727095 TI - Simultaneous quantitation of cytokine mRNAs by reverse transcription-polymerase chain reaction using multiple internal standard cRNAs. AB - Cytokines produced in abnormal amounts or patterns contribute to many immunologically mediated human diseases. We describe a competitive reverse transcription-polymerase chain reaction (RT-PCR) assay to measure interleukin (IL)1-2, IL-4, and interferon-gamma (IFN-gamma) mRNAs within the sample. Internal standard cRNAs and native cytokine mRNAs are reverse transcribed and then amplified by PCR in the same reaction tubes to control for tube-to-tube variability in these reactions. In contrast to systems that use a single multigene internal standard cRNA, this method uses separate internal standard cRNAs for IL-2, IL-4, and IFN-gamma, allowing independent dosing of the internal standards, which reduces the number of tubes processed and the amount of starting mRNA required. Internal standards are produced from cytokine cDNAs by the insertion of short segments of DNA. The same oligonucleotide primers are used to amplify internal standard and native cytokine cDNAs. Each internal standard cDNA and its matching native cytokine cDNA are amplified with equal efficiency. The RT PCR products of the internal standards and native cytokines are distinguished by size. This technique can detect a twofold difference in mRNA levels. Examples of using this technique to measure cytokine mRNAs in peripheral blood mononuclear cells and in bronchoalveolar lavage cells are given. PMID- 8727096 TI - Reverse transcriptase-polymerase chain reaction amplification of MDR1 gene expression in adult soft tissue sarcomas. AB - Expression of the multidrug resistance gene MDR1 is reported to be an important determinant of the response to chemotherapy and survival in some cancers. We compared three methods for determining the intrinsic MDR1 expression in soft tissue sarcomas. We studied MDR1 gene expression in 39 samples from 33 cases of soft tissue sarcomas comprising 11 liposarcomas, nine malignant fibrous histiocytomas, six leiomyosarcomas, four malignant schwannomas, three fibrosarcomas, three synovial sarcomas, and three epithelioid sarcomas, and seven cases of benign soft tissue tumors in adult patients. To detect MDR1 mRNA, reverse transcriptase-polymerase chain reaction (RT-PCR) was performed in all samples. Furthermore, RNA dot-blot analysis with digoxigenin-labeled RNA probe and immunohistochemistry with JSB-1 and C-219 antibodies for P-glycoprotein were employed in 34 and 37 samples in soft tissue sarcomas, respectively. We compared these three detection techniques. Of the 39 specimens, 18 (46%) showed MDR1 PCR products. Liposarcomas (six of 11), malignant fibrous histiocytomas (six of nine), leiomyosarcomas (four of six), fibrosarcomas (two of three) revealed high or intermediate MDR1 expression at high frequency. No MDR1 expression was detectable in malignant schwannomas, synovial sarcomas, or epithelioid sarcomas. Of seven benign soft tissue tumors, one ganglioneuroma and one lipomatosis showed low levels of MDR1 expression. By RNA dot-blot analysis, MDR1 transcripts were detectable in 12 of 34 specimens (35%). Four samples were negative by dot blot despite positivity with RT-PCR. Concordance between MDR1 expression by RNA level with RT-PCR and dot blot and at the protein level with immunohistochemistry using C-219 was found in 16 (47%) of the 34 comparable specimens. Eight samples showed positive immunoreactivity for C-219 despite negative results in RT-PCR and dot blot analysis. The intrinsic MDR1 expression in soft tissue sarcoma seemed to depend on certain tumor types, such as liposarcoma, malignant fibrous histiocytoma, leiomyosarcoma, and fibrosarcoma. For the evaluation of MDR1 expression, RT-PCR is useful because of its relative simplicity and sensitivity. However, the clinical significance of such low levels of MDR1 expression detected only by RT-PCR must be discussed within systematically treated patient groups. PMID- 8727097 TI - Detection of t(11;22)(q24;q12) translocation breakpoint in paraffin-embedded tissue of the Ewing's sarcoma family by nested reverse transcription-polymerase chain reaction. AB - Tumors of the Ewing's sarcoma family often present a major diagnostic challenge for the pathologist. In recent years, significant progress has been made in identifying characteristic chromosomal rearrangements associated with certain solid tumors. More than 85% of Ewing's sarcoma and related tumors present a specific t(11;22) (q24;q12) balanced translocation, which generates a fusion transcript of the EWS gene and the FLI-1 gene. The cloning of the t(11;22)(q24;q12) breakpoint has raised the possibility of using a reverse transcription-polymerase chain reaction (RT-PCR) based assay as a diagnostic tool. We report an improvement of the established method, which currently depends on fresh or snap-frozen tissue, so that it is possible to use formalin-fixed, paraffin-embedded tissue as a source of RNA. The described nested RT-PCR assay enables the pathologist to investigate retrospectively archival tumor samples or to confirm the diagnosis in cases where no fresh or frozen material is available. PMID- 8727098 TI - Semi-automated ELISA-based detection system for verifying the authenticity of amplified t(14;18)-containing products. AB - PCR product sizing on ethidium bromide-stained gels, coupled with Southern transfer and hybridization with nonisotopic probes, is an effective way of detecting t(14;18)(q32;q21). We evaluated an alternative ELISA-based test for detecting amplified t(14;18) products. Digoxigenin (DIG)-labeled dUTP is incorporated in a standard PCR method for amplification of bcl-2 major breakpoint region (mbr) rearrangements. The product is hybridized to a specific biotinylated DNA probe internal to the mbr primer, placed in streptavidin-coated wells of a microtiter plate, and detected with a alkaline phosphatase-conjugated anti-DIG antibody and enzyme substrate (pNpp). The colorimetric product is quantitated by an automated optical density (O.D.) reader. We evaluated 13 mbr-positive follicular lymphomas (FL), five mbr-negative B-cell neoplasms (BCN), 16 reactive lymphoid hyperplasias (RLH), 14 cases of Hodgkin's disease (HD), and normal peripheral blood samples from 20 healthy volunteers. All samples were evaluated in duplicate on separate plates. Positive [t(14;18)-containing cell line] and negative [cell line without t(14;18); master mix only] controls, and a standard curve were included with each run. Numerical O.D. readings from the specific hybridization assays revealed differences between FL and the other categories. All FL had an O.D. reading at > 2.0. The vast majority of RLH, HD, BCN, and normal peripheral blood samples showed O.D. readings well below 2.0. Specifically, 13/16 RLH and all HD, BCN, and normal peripheral blood samples had an O.D. of < or = 0.6 in all runs. The three outliers, which were all < 2.0, may represent the low level detection of t(14;18)-containing cells in RLH similar to previous reports. Moreover, all but four RLH had O.D. readings above the background negative controls, suggesting that rare t(14;18)-containing cells may have been present in these samples, as well. Dilution studies estimate that this assay is capable of detecting 1 t(14;18)-containing cell in approximately 10(5) cells, a greater level of sensitivity than can be obtained with gel visualization alone. We conclude that this semi-automated, potentially quantifiable ELISA-based system is a useful, objective and reproducible alternative hybridization procedure for verifying PCR product specificity in this setting. PMID- 8727099 TI - A rapid polymerase chain reaction-based technique for detecting clonal T-cell receptor gene rearrangements in cutaneous T-cell lymphomas of both the alpha beta and gamma delta varieties. AB - T-cell receptor-gamma gene rearrangements provide specific clonal markers for a variety of lymphoid malignancies. T-cell receptor gene rearrangements in patients with cutaneous T-cell lymphoma were examined using conventional Southern blot analysis and a newly developed polymerase chain reaction (PCR)-based technique. The oligoprimers amplified a rearranged V gamma and J gamma segment (including the N region) of the T-cell receptor-gamma gene, and products were resolved using high-resolution nondenaturing polyacrylamide gel electrophoresis. Our results demonstrated concordance between the two techniques in 10 patients with cutaneous T-cell lymphomas (including nine cases of C beta and one case of delta 2 TCR gene rearrangements) and 10 negative controls. In the present study, we have shown that this PCR-based method provides a highly sensitive, specific technique for the detection of T-cell clones of both the alpha beta and gamma delta varieties and could be used in both fresh and formalin-fixed, paraffin-embedded tissues. It is estimated that this PCR-based technique is 10 to 50 times more sensitive than conventional Southern blot analysis in the detection of small T-cell clones. PMID- 8727100 TI - Integrin alpha V beta 3 expression by bone-residing breast cancer metastases. AB - Breast cancer metastasis to bone is a multistep process requiring attachment of tumor cells to the bone and bone marrow environment. The precise adhesion molecules involved in skeletal homing of breast cancer to bone are unknown but likely include integrins. We investigated the expression of vitronectin receptor (alpha V beta 3) by breast cancer cells residing in bone because this heterodimer mediates osteoclast-bone recognition. We used immunohistochemistry and in situ hybridization in a systematic study of 22 bone biopsies containing breast cancer metastases and available samples of corresponding primary tumors and normal breast and compared alpha V beta 3, alpha 2 beta 1, and alpha B beta 5 integrin expression. The results showed that alpha V beta 3 was strongly expressed by normal breast epithelium and was decreased in some and strongly expressed in other primary invasive breast carcinomas. In contrast, this integrin heterodimer was abundant in all breast cancer cells metastatic to bone. In situ hybridization revealed high levels of steady-state mRNA corresponding to sites of protein expression; alpha 2 beta 1 was weakly expressed in both primary and metastatic tumors, and alpha V beta 5 was not detected. Our results showed an overexpression of alpha V beta 3 by bone-residing breast cancer cells and suggest either subclonal selection of alpha V beta 3-expressing tumor cell populations or upregulation of alpha V beta 3 in the bone microenvironment. PMID- 8727101 TI - Improved detection of human papillomavirus infection in genital intraepithelial neoplasia in human immunodeficiency virus positive (HIV +) women by polymerase chain reaction-in situ hybridization. AB - The prevalence of genital human papillomavirus (HPV) infection was evaluated in 30 consecutive human immunodeficiency virus (HIV) + women by polymerase chain reaction (PCR)-in situ hybridization (ISH) on paraffin-embedded tissue sections and compared with that found with standard ISH. Biopsies were removed from normal or neoplastic areas in the cervix, vagina, and vulva, and ISH was performed with biotinylated or fluorescein isothiocyanate genomic DNA probes. One probe was used for HPV screening and others for HPV typing (types 6, 11, 16, 18, 31, and 33). Sequences were amplified by the "hot-start" PCR method and followed by standard ISH. Among the 30 HIV + women, 90% scored HPV + in one or several locations by PCR-ISH, whereas only 67% were positive by ISH. Oncogenic HPV types were found in 63% by PCR-ISH and in only 43% by ISH. The same HPV types detected by standard ISH were also recognized by PCR-ISH, but with the latter the signal was amplified. Moreover, some HPV types were found with PCR-ISH but not by ISH. We conclude that PCR-ISH is a valuable and sensitive method for specific detection of HPV. PMID- 8727102 TI - A polymerase chain reaction-based assay for the rapid detection of gene amplification in human tumors. AB - A differential polymerase chain reaction (PCR) protocol was established for semiquantitative, nonradioactive detection of gene amplification using a DNA sequencer. Oncogene fragments and control DNA sequences were simultaneously PCR amplified using fluorescent-labelled primers. Analysis of the PCR products allowed quantitative assessment of gene copy numbers in this coamplification assay. Using this approach, we examined a series of 132 brain tumors for amplification of the epidermal growth factor receptor (EGFR) gene. The same set of tumors was also analyzed by Southern blotting and hybridization with a radiolabelled EGFR probe. Both methods yielded virtually identical results. This technique has a great potential for nonradioactive screening of large tumor panels of oncogene amplification. PMID- 8727103 TI - Aspects of adherence of oral spirochetes. AB - Oral spirochetes are present in the oral cavity in various numbers and forms and have been strongly implicated as playing a role in the etiology of periodontal disease. Because adherence to host tissues is a critical first step in establishing a bacterial infection, reports on the attachment of oral spirochetes to host tissues and matrix components were reviewed. The great bulk of the literature concerns Treponema denticola; however, where there is information regarding other oral spirochete species, it will be noted. PMID- 8727104 TI - Setting reactions in dental amalgam. Part 1. Phases and microstructures between one hour and one week. AB - The literature on the setting mechanisms of dental amalgams made from powders of silver-rich alloys of tin and/or copper has been critically reviewed. In Part 1 of the review, the microstructure and phase content of recently set amalgams are described. The composition, morphology, and location of product phases are emphasized, since these features are clues to the setting reaction. Thus, Part 1 provides the background needed to understand the kinetics of the setting reactions, which is the topic of Part 2 of the review. PMID- 8727105 TI - Setting reactions in dental amalgam. Part 2. The kinetics of amalgamation. AB - The literature on the setting mechanisms of dental amalgams made from powders of silver-rich alloys of tin and/or copper has been critically reviewed. Part 2 is a review of the kinetics of the reactions that convert the mixture of alloy powder and liquid mercury to hardened amalgam containing the phases and microstructures described in Part 1. It is emphasized that amalgamation is a non-equilbrium process in which hardened microstructures are determined as much by kinetics as by chemistry. The setting reaction begins with dissolution of silver and tin into liquid mercury; most of the product phases precipitate in the liquid mercury. The processes that produce supersaturation in the liquid mercury and the subsequent nucleation and growth of solid phases are considered. Mass balance relationships that provide insight into the factors that control the volume fraction of the undesirable gamma 2 Sn-Hg phase are described. The nucleation and growth of eta' Cu-Sn crystals are also discussed; it is found that these crystals nucleate on copper-rich phases and grow into the liquid mercury. Finally, aspects of the setting reaction that are controlled by intergranular and interphase diffusion in the solid are discussed. These aspects include: the supersaturation of silver and tin within the liquid mercury, nucleation and growth of the beta 1 Ag-Hg phase in the surfaces of alloy particles, and the decomposition of initially formed gamma 2 Sn-Hg. PMID- 8727106 TI - Oral mucosal Langerhans' cells. AB - Langerhans' cells (LC) are dendritic, antigen-presenting cells present within the epithelium of skin and mucosa, including that of the oral cavity. This article reviews the literature on the phenotypic and functional features of oral mucosal Langerhans' cells, and speculates on other aspects by extrapolating from data on their epidermal counterparts. PMID- 8727107 TI - T-lymphocyte interactions with endothelium and extracellular matrix. AB - T-lymphocyte movement out of the bloodstream and into tissue is critical to the success of these cells in their role in immunosurveillance. This process involves interactions of the T-cell with endothelium as well as with extracellular matrix. Central to these interactions are a number of T-cell adhesion molecules and their endothelial and extracellular matrix ligands. The identification and functional characterization of adhesion molecules have been the subject of intensive research in recent years. We highlight here the latest developments in this rapidly expanding field as they pertain to T-cell interactions with endothelial cells and extracellular matrix components, including: (1) identification of adhesion molecule families, including the selectins, mucins, integrins, immunoglobulin superfamily members, and cadherins; (2) elucidation of the multi step adhesion cascade that mediates the rolling, arrest, and eventual diapedesis of T-cells through the vascular endothelium into the surrounding tissue; (3) the changes in adhesion molecule expression that accompany T-cell maturation and activation, and the impact of those changes on T-cell migration; (4) the functional relevance of the extracellular matrix for T-cell function; and (5) the clinical relevance of adhesion molecules and the potential for targeting these molecules for the amelioration of immune-mediated diseases. PMID- 8727108 TI - A critical evaluation of methods for root coverage. AB - Gingival recession is the exposure of root surfaces due to apical migration of the gingival tissue margins. Although it seldom results in tooth loss, marginal tissue recession is associated with thermal and tactile sensitivity, esthetic complaints, and a tendency toward root caries. This article reviews the current surgical procedures for the coverage of exposed root surfaces, including their advantages and disadvantages. Today, the outcome of root coverage procedures is predictable, aesthetically acceptable, and, together with the use of guided tissue regeneration, at the forefront of regenerative procedures. PMID- 8727109 TI - Leprosy neuropathy: correlation of clinical and electrophysiological tests. AB - This report describes the neurological and electrophysiological examination of 35 subjects with leprosy (average duration of symptoms 3.4 years, average time since diagnosis 7.5 months). Clinical examination in the distribution of non-dominant median and ulnar nerves was performed with the following clinical methods: touch sensation with 0.05 gm. Monofilament nylon, thermal sensation with a thermal sensitivity testing device, voluntary muscle testing and nerve palpation. At least one abnormality was found in 22 ulnar and 13 median nerves (63% and 37%, respectively). Nerve palpation was the most frequent clinical abnormality, while the other methods had similar frequencies of abnormality. Electrophysiological studies were performed on the ipsilateral side of the leprosy subjects and on 32 age-matched normal subjects. Electrophysiological responses from the leprosy subjects were evaluated by criteria established from normal subject data. Abnormal or absent responses were found in 21/35 ulnar sensory, 12/35 ulnar motor, 9/35 median sensory and 6/35 median motor responses among the leprosy subjects. The most important electrodiagnostic findings were: (i) low sensory amplitudes and (ii) drops in amplitude and NCV over the across-elbow segment of the ulnar nerve. Both clinical and nerve conduction abnormalities were positively associated with duration of leprosy symptoms. The four clinical methods were compared for concordance with nerve conduction data by cross-tabulation. The two sensory measures, monofilaments and the thermal sensitivity device, had the highest concordances. Usefulness of clinical tests for nerve damage in leprosy may vary depending on whether the purpose is for diagnosis, patient education or clinical follow-up. PMID- 8727111 TI - Pregnancy and leprosy neuropathy. AB - Women with leprosy (even apparently cured) run a serious risk of deterioration in nerve function when they become pregnant. During pregnancy and lactation the woman with leprosy may suffer: relapse, reactivation and transient exacerbation maximally in late pregnancy; ENL in the first and third trimesters, continuing with nerve damage postpartum; RR maximally postpartum, even after MDT and RFT; neuritis affecting almost 50% of women in any pregnancy/lactation, in most cases as "silent" neuritis with new motor and sensory loss, even after MDT-RFT, and stocking-and-glove anaesthesia even in PB women and post MDT-RFT. Those incubating the infection develop overt disease frequently in reaction. This tragic cycle can only be stopped by a combination of: (i) leprologists and leprosy control personnel understanding the problems of leprosy in pregnant and lactating mothers; (ii) well-planned health education for leprosy patients, and both leprosy and maternal health care workers and (iii) the highest standard of clinical supervision during pregnancy, prolonged lactation and at regular intervals during the woman's reproductive life, even after she would normally be released from surveillance after completion of multiple drug treatment (MDT). PMID- 8727110 TI - Leprous neuropathy as an autoimmune phenomenon. PMID- 8727112 TI - Neuritic leprosy. PMID- 8727113 TI - Nerve in reversal reaction. AB - 1. Much of the nerve destruction in leprosy takes place during the reactive phase, both during ENL reaction and RR. 2. The high risk patients expected to develop RR are borderline patients with generalized lesions (more than 10 skin lesions) and those presenting with three or more thickened nerve trunks. 3. In RR there is a sudden enhancement of already existing DTH to M. leprae and its antigens resulting in the release of excessive quantities of TNF alpha, INF gamma, and IL-2. The triggering mechanisms of this phenomenon is poorly understood. 4. The already existing granulomas suddenly increase considerably in size due to oedema and rapid influx of lymphocytes, Langhan's and foreign body giant cells. Fragments of M. leprae are also present in the granuloma of some patients. 5. In RR, the acute granulomatous inflammation can produce destruction of nerves even to the extent of causing caseous necrosis of the nerve tissue and irreversible paralysis. The swelling of the nerves due to sudden increase in inflammatory cells and oedema within an unyielding perineurium produce ischaemia and transient paralysis. 6. With prompt administration of anti-inflammatory drugs, paralysis recovers quickly, if it is of ischaemic origin; but will not recover if the Schwann cells and other nerve tissues are destroyed as a result of the immune granuloma. 7. A course of corticosteroids for six months along with anti-leprosy therapy is suggested in high risk patients as a preventive measure. 8. Further the serious problem of continuing nerve damage after clinical cure should be urgently tackled. PMID- 8727114 TI - Autonomic nerve affection in leprosy. AB - Leprosy has been shown to affect almost all systems of human body and abnormalities in functions of autonomic nerves innervating various parts have been observed in several studies. In the skin and its appendages, the common changes are anhidrosis and varying degree of impaired sweat response. Signs of denervation of iris and reduced intraocular pressure are permanent features of autonomic involvement in the eye. In the cardiac autonomic functions, rhythm disturbances have been documented by several investigators. Respiratory function test studies have shown impaired breath holding time and decreased response to cough as well as other changes indicating blockade of vagus nerves and sympathetic plexus. Abnormal testicular pain sensation and diminished nocturnal penile tumiscence provide evidence of afflication of autonomic nerves of male genital system. Other important autonomic nervous system involvements include the nerves innervating the capillaries of legs. These changes have been observed to be more in extensive and long standing disease which indicate the need to study all these aspects in prospective studies specially in the light of early institution of multidrug treatment. PMID- 8727115 TI - Role of surgery of nerves in leprosy in the restoration of sensibility in hands and feet of leprosy patients. PMID- 8727116 TI - Nerve tissue culture as an approach towards the study of onset of reactions in the nerve in leprosy--observations and hypothesis. PMID- 8727117 TI - Posterior tibial neurovascular decompression for restoration of plantar sweating and sensibility. PMID- 8727118 TI - Peripheral nerve reconstruction using denatured muscle autografts for restoring protective sensation in hands and feet of leprosy patients. PMID- 8727119 TI - The leprosy granuloma in nerve: summary of recent work. PMID- 8727120 TI - Animal model for leprous neuropathy: our contribution. PMID- 8727121 TI - A semi quantitative analysis of bacterial load in different cell types in leprous nerves using transmission electron microscope. AB - Thirty lepromatous (BL-LL) and 25 tuberculoid (TT-BT) nerve lesions obtained from untreated cases of leprosy were scanned using transmission electron microscope for assessing the bacterial load in different cell types. Major bulk of infection was seen in the Schwann cells of nonmyelinated fibres, in both early lepromatous and tuberculoid nerve lesions, suggesting that M. leprae spread mainly via the Schwann cells within the nerve. PMID- 8727122 TI - Cytological needle aspiration for the diagnosis of pure neural leprosy. PMID- 8727123 TI - Host parasite response in nerve in leprosy. AB - Nerve granulomas occur at all points across the leprosy spectrum. Studies have been made using experimental models in which mycobacteria were injected directly in the sciatic or posterior tibial nerve of the guinea pig. Clinical and electrophysiological studies demonstrated axonal damage which was confirmed by morphometric studies showing disrupted myelin sheaths and in places complete demyelination. Further immunohistological studies showed a complete disappearance of staining for certain neuropeptides. The role of Schwann cells has also been investigated. Schwann cells in nerves affected by mycobacterial granulomas, both experimental and in leprosy patients were not demonstrated to be MHC class II positive suggesting that they did not play a role in antigen presentation. Macrophages in leprosy granulomas were shown to contain TNF alpha, suggesting that this cytokine played a role in axonal damage. The role of mycobacterial heat shock protein in nerve granulomas has not as yet been determined. The localized nature of granulomas in leprosy nerves and nerves with experimental mycobacterial granulomas has been studied by a process of excision and repair with muscle grafts. Marked recovery has been demonstrated by clinical, electrophysiological, morphometric and immuno-histochemical techniques, the latter demonstrating a return of neuropeptide production. PMID- 8727124 TI - Assessment of nerve function under field conditions and its usefulness in leprosy. PMID- 8727125 TI - Historical aspects of the rickettsioses. PMID- 8727126 TI - Rickettsial microbiology. PMID- 8727127 TI - Rocky Mountain spotted fever. PMID- 8727128 TI - Other spotted fevers. PMID- 8727129 TI - The typhus group. PMID- 8727130 TI - Rickettsialpox. PMID- 8727131 TI - Q fever and trench fever. PMID- 8727132 TI - Ehrlichial infections. PMID- 8727133 TI - Bacillary angiomatosis. PMID- 8727134 TI - Laboratory diagnosis of rickettsial diseases. PMID- 8727135 TI - Magnetic resonance image-guided stereotactic cingulotomy for intractable psychiatric disease. AB - We describe the modern operative technique of magnetic resonance (MR) image guided stereotactic cingulotomy and discuss the indications, results, and complications of this procedure. A retrospective analysis of psychiatric outcome was performed for 34 patients with intractable major affective disorder and/or obsessive-compulsive disorder who underwent MR image-guided stereotactic cingulotomy since 1991. Fourteen patients underwent multiple cingulotomies (50 total procedures). Overall, 38% of the patients were classified as responders, 23% as possible responders, and 38% as nonresponders. Of the patients who did not respond to initial cingulotomies and who underwent multiple cingulotomies, 36% became responders, 36% possible responders, and 28% nonresponders. There were no deaths or long-term side effects related to the procedure. The therapeutic results of MR image-guided stereotactic cingulotomy are similar to the results of earlier methods of cingulotomy, and the use of MR imaging offers substantial technical advantages. This procedure also compares favorably with other neurosurgical procedures performed for intractable psychiatric disease with a low rate of undesired side effects. Cingulotomy is safe and well tolerated, with over one-third of the patients demonstrating significant improvement; however, prospective long-term follow-up studies are needed to further define the role of surgery in treating intractable psychiatric disease. PMID- 8727136 TI - Surface and superficial surgical anatomy of the posterolateral cranial base: significance for surgical planning and approach. AB - We have performed an anatomic study, 15 using fixed cadaveric preparations, with the goal of identifying surface landmarks that will reliably locate the underlying transverse and sigmoid sinus complex. Simple morphometric relationships were first determined on both sides of each specimen to yield 30 sides measured. The following relationships were determined: 1) zygoma root asterion, 2) asterion-mastoid tip, 3) zygoma root-suprameatal spine (Henle's spine), 4) asterion-suprameatal spine, 5) mastoid tip-suprameatal spine. The relationship of the asterion to the transverse-sigmoid junction was determined by bone removal. Also, the distances from the asterion to the sigmoid sinus-superior petrosal sinus junction and the superior margin of the transverse sinus were studied. Surface and marks were found to have definitive relationships to underlying anatomic substrates in all specimens studied. The critical relationships that were concluded from this study can be described in terms of two easily identified lines between bony surface structures. A line drawn from the zygoma root to the inion, i.e., the superior nuchal line, reliably located the rostrocaudal level of the transverse sinus in all specimens. Although the asterion did not consistently fall on this line, the transverse-sigmoid junction could reliably be placed at the anteroposterior level of the asterion. Further, a line drawn from the squamosal-parietomastoid suture junction to the mastoid tip reliably defined the axis of the sigmoid sinus through the mastoid. We also found that the junction of the squamosal and parietomastoid sutures lay over the anterior border of the upper curve of the sigmoid sinus. The anterior portion of the supramastoid crest correlated with the level of the middle fossa. These surface relationships all have significance for posterolateral approaches to the cranial base. Since performing this study, these relationships have been found reliable for operative planning in our clinical cases. PMID- 8727137 TI - Cerebral hyperemia after arteriovenous malformation resection is related to "breakthrough" complications but not to feeding artery pressure. The Columbia University Arteriovenous Malformation Study Project. AB - To study the pathophysiology of idiopathic postoperative brain swelling or hemorrhage after arteriovenous malformation resection, termed normal perfusion pressure breakthrough (NPPB), we performed cerebral blood flow (CBF) studies during 152 operations in 143 patients, using the xenon-133 intravenous injection method. In the first part of the study, CBF was intraoperatively measured (isoflurane/N2O anesthesia) during relative hypocapnia in 95 patients before and after resection. The NPPB group had a greater increase (P < 0.0001) in mean +/- standard deviation global CBF (28 +/- 6 to 47 +/- 16 ml/100 g/min, n = 5) than did the non-NPPB group (25 +/- 7 to 29 +/- 10 ml/100 g/min, n = 90); both arteriovenous malformation groups showed greater increase (P < 0.05) than did controls undergoing craniotomy for tumor (23 +/- 6 to 23 +/- 6 ml/100 g/min, n = 22). Ipsilateral and contralateral CBF changes were similar. In a second cohort of patients with arteriovenous malformations, CBF was measured at relative normocapnia and it increased (P < 0.002) from pre- to postresection (40 +/- 13 to 49 +/- 15 ml/100 g/min, n = 57). There were no NPPB patients in this latter cohort. The feeding mean arterial pressure was measured intraoperatively before resection or at the last embolization before surgery (n = 64). The feeding mean arterial pressure (44 +/- 16 mm Hg) was 56% of the systemic arterial pressure (78 +/- 12 mm Hg, P < 0.0001) and was not related to changes in CBF from pre- to postresection. There was an association between increases in global CBF from pre- to postresection and NPPB-type complications, but there was no relationship of these CBF changes to preoperative regional arterial hypotension. These data do not support a uniquely hemodynamic mechanism that explains cerebral hyperemia as a consequence of repressurization in hypotensive vascular beds. PMID- 8727139 TI - Management and long-term outcome of adenoid cystic carcinoma with intracranial extension: a neurosurgical perspective. AB - Adenoid cystic carcinoma is a tumor of minor and major salivary glands that often invades the cranial base and intracranial cavity via local and perineural spread. In the past, the role of neurosurgeons in managing these tumors has been limited. The growth of interdisciplinary cranial base surgical approaches has now increased the involvement of neurosurgeons in the management of these tumors. We present a series of 16 patients with adenoid cystic carcinomas with cranial base and intracranial extension, the largest series reported in the neurosurgical literature. We have focused our approach on a neurosurgical perspective and made recommendations for the treatment of these tumors in relation to the following specific aspects of this disease. 1) The management of the carotid artery: In our experience, when the carotid artery is involved by tumor, a preliminary cerebral revascularization procedure with a cervical carotid to middle cerebral artery vein bypass graft should be performed before tumor resection. 2) The management of the cavernous sinus and orbit: Cavernous sinus tumor should be removed as fully as possible, but every effort should be made to preserve the IIIrd and IVth cranial nerves to achieve optimal functional and cosmetic results. The orbit should be exenterated when there is intraconal involvement; otherwise, intraorbital tumor can be removed with orbital preservation. 3) The use of palliative surgery: We have found that the use of palliative surgery can be considered even in patients whose extent of local disease precludes a surgical cure. The slow progression of the disease allows for long-term survival of many patients with advanced local disease and even of those with metastatic disease. 4) The long-term survival of these patients: In our series, six patients had no evidence of local disease and a mean survival of 72 months, one living patient had evidence of local disease and has survived 56 months, eight patients died of disease, with a mean survival of 137 months, and one patient died of complications after surviving for 63 months. PMID- 8727138 TI - Long-term treatment of malignant gliomas with intramuscularly administered polyinosinic-polycytidylic acid stabilized with polylysine and carboxymethylcellulose: an open pilot study. AB - Polyinosinic-polycytidylic acid stabilized with polylysine and carboxymethylcellulose (poly-ICLC) (10-50 mcg/kg, administered intramuscularly one to three times weekly) was given for < or = 56 months to 38 patients with malignant gliomas. There was minimal or no toxicity. Twenty of 30 patients (66%) receiving at least twice weekly poly-ICLC showed regression or stabilization of gadolinium-enhancing tumor, as revealed by magnetic resonance imaging (median = 65% volume decrease). All but one patient with anaplastic astrocytomas who received continuous poly-ICLC remain alive, with a median progression-free survival of 54 months from diagnosis. Median Kaplan-Meier survival is 19 months for patients with glioblastomas who receive at least twice weekly poly-ICLC treatments. Tumor response was associated with 2',5' -oligoadenylate synthetase activation (P = 0.03) but not with serum interferon. We hypothesize clinical activation by poly-ICLC of a basic host tumor suppressor system. Prolonged, quality survival with tumor stabilization or regression confirmed by magnetic resonance imaging for most patients with anaplastic astrocytomas and glioblastomas suggests that more extensive laboratory and controlled clinical studies are warranted. The concept of long-term, broad spectrum stimulation of host defenses with nontoxic, inexpensive double-stranded ribonucleic acids, such as low-dose poly-ICLC, may be applicable to the treatment of other malignancies. PMID- 8727140 TI - Prognostic significance of type 1 neurofibromatosis (von Recklinghausen Disease) in childhood optic glioma. AB - Although the association between optic glioma and neurofibromatosis is well recognized, few studies have systematically compared the outcomes of patients with optic gliomas and neurofibromatosis and patients with optic gliomas without neurofibromatosis. In the present study, patients with optic gliomas and Type 1 neurofibromatosis (NF-1) were compared with patients with optic gliomas without NF-1, with respect to survival, time to tumor progression, and tumor location. Forty-four patients with optic gliomas who were evaluated between 1949 and 1991 were studied retrospectively. Sixteen of 44 patients (36%) met the National Institutes of Health criteria for NF-1. The medical records of all patients were examined, and letters of inquiry were sent to every living patient to ascertain current health statuses. Death certificates were obtained to determine causes of death. Follow-up averaged 7.2 years (10.2 yr for patients with NF-1, 5.4 yr for patients without NF-1). The 5- and 10-year survival rates for patients with optic gliomas and NF-1 were 93 and 81%, respectively. For those patients with optic gliomas who did not have NF-1, 5- and 10-year survival rates were 83 and 76%, respectively. Seventeen patients experienced tumor progression (5 with NF-1, 12 without NF-1). A difference was observed in the mean time to tumor progression (first relapse) between the two groups (mean time with NF-1, 8.37 yr; without NF 1, 2.39 yr [P < 0.01]). However, no significant difference in overall survival, as evaluated by a log-rank test of the respective Kaplan-Meier survival curves, was observed between the two groups. A significant difference in distribution of tumor location between the group with NF-1 and the group without NF-1 was also noted (Fisher's exact test, P = 0.0338), although the number of patients evaluated in this series was too small to determine whether this difference in tumor location influenced relapse rate. We conclude that optic gliomas in patients with neurofibromatosis have a different distribution of location as opposed to those in patients without neurofibromatosis, and, for first relapse, the presence of neurofibromatosis is a significant favorable factor. PMID- 8727141 TI - The relevance of hemodynamic factors to perioperative ischemic complications in childhood moyamoya disease. AB - Of 124 children younger than 15 years who underwent surgery for moyamoya disease, 21 (16.9%) experienced perioperative ischemic complications that could not be unequivocally attributed to the surgery. Eleven of the 21 patients experienced infarctions, and 10 experienced reversible ischemic neurological deficits without new lesions, as revealed by computed tomographic scans. An examination of the patients' perioperative clinical and laboratory data revealed that the mean values of intra- and postoperative minimum arterial carbon dioxide pressure, maximum arterial carbon dioxide pressure, and mean arterial pressure were similar in patients with and without ischemic complications. However, in patients with perioperative complications, the incidence of preoperative transient ischemic attacks (TIAs) and intra- and postoperative hypercapnia (maximum arterial carbon dioxide pressure > 45 mm Hg) was significantly higher. In addition, 7 of the 11 perioperative infarctions occurred in patients with frequent preoperative TIAs and intra- and postoperative hypercapnia. Cerebral blood flow studies with preoperative acetazolamide loading showed that the new infarctions were located in areas in which the cerebral blood flow had been compromised. Our results suggest that the occurrence of frequent preoperative TIA is an important indicator of the instability of the cerebral hemodynamics and of the risk of perioperative ischemic complications. To prevent these complications, preoperative management aimed at stabilizing the hemodynamic status is very important. Children who have moyamoya disease and who experience frequent preoperative TIAs are at risk for ischemic brain damage caused by hypercapnia as well as hypocapnia and hypotension. The establishment and maintenance of normocapnia with normotension are highly desirable for the perioperative management of moyamoya disease in children. PMID- 8727142 TI - Radiofrequency lesion adjacent to the dorsal root ganglion for cervicobrachial pain: a prospective double blind randomized study. AB - Each of 20 consecutive patients with a history of at least 1 year of intractable chronic cervicobrachial pain was randomly assigned to one of two treatment groups. The pretreatment investigation included at least three diagnostic segmental nerve blocks in each patient. Each patient in Group 1 received a 67 degrees C radiofrequency lesion adjacent to the dorsal root ganglion. The patients in Group II were treated in an identical manner as those in Group I, except that no actual radiofrequency lesion was made. Neither the therapist nor the patients were aware of the treatment group assignment. All patients were questioned about their pain experience 1 week before and 8 weeks after the procedure. The following tests were used in evaluating patient response: Visual Analogue Scale (VAS); McGill Pain Questionnaire, Dutch Language Version (MPQ DLV); and Multidimensional Pain Inventory, Dutch Language Version (MPI-DLV). These tests showed that 8 weeks after the procedure, there was a significant number of "successful" patients in Group I compared to Group II (P = 0.0027); there was a significant reduction in VAS score (P < 0.01) and also in parameters measured with MPQ-DLV and MPI-DLV in Group I. This study indicates that a 67 degrees C radiofrequency lesion adjacent to the dorsal root ganglion can result in a significant alleviation of pain in chronic cervicobrachial pain. PMID- 8727143 TI - Surgical treatment of nonprogressive neurological deficits in children with sacral agenesis. AB - I report my experience with 10 children with varying degrees of sacrococcygeal agenesis. There were four children with total agenesis and four with partial agenesis. All of these children had neurological deficits that had been static since birth. Radiological evaluation of these children revealed the presence of tethered cord in two children and tethered cord with lipomeningocele in one. Surgical correction of these intraspinal anomalies led to the improvement of urinary incontinence in these children. This report highlights the fact that children with sacral agenesis and nonprogressive neurological deficits may have correctable intraspinal anomalies, and hence all of the children with sacral agenesis should be evaluated for the presence of treatable intraspinal anomalies. PMID- 8727144 TI - Quantitative three-dimensional anatomy of the subaxial cervical spine: implication for anterior spinal surgery. AB - Knowledge of the quantitative anatomy of the subaxial cervical vertebrae is essential to safely perform anterior cervical surgery and to ensure adequate decompression of neural structures. In spite of this, little has been published in the neurosurgical literature regarding the spatial relationship of the lower cervical vertebrae and the implications of this anatomy for anterior cervical surgery. We report the three-dimensional analysis of the mid- and lower cervical spine in 10 cadaver specimens for 50 disarticulated vertebrae and discuss the relevance of this analysis to surgery in this region. Measurements were made using real-time video analysis of images transferred from a Zeiss microscope equipped with an image splitter and a Sony charge-coupled device camera. Images were then transferred to an IBM personal computer-based image analysis system. Analysis of variance was used to test for significant differences among the C3-C7 vertebral measurements. Important relationships of the vertebral artery to the anteroposterior diameter of the vertebral bodies and its variations from C3 to C7 are discussed. The vertebral artery migrates posteriorly to anteriorly from C3 to C6 and posteriorly again at C7; the implications of these variations are discussed for decompression of the neural foramen. Another finding showing that the inter-Luschka distance increases from C3 to C7 is important for adequate lateral decompression in anterior cervical spinal surgery. The pedicle to the Luschka joint was measured at the different levels; the pedicle is lateral to the Luschka joint from C3 to C6 and medial to the joint at C7. This variation explains the lack of root decompression at some cervical levels, even when decompression extends to the Luschka joint bilaterally. We also report the measurements of the vertebral bodies and the spinal canal and compare the results with other published data. We think these measurements provide guidelines for operating on the anterior cervical spine, facilitate adequate decompression of the spinal cord and neural foramen, and increase the margin of safety of the surgeon. PMID- 8727145 TI - Bypass coaptation procedures for cervical nerve root avulsion. AB - In the past, patients with cervical spinal nerve root avulsions were resigned to accept a natural crippling from upper extremity neurological deficits. Recently, bypass coaptation procedures have resulted in functional return of denervated muscles after such avulsions, much to the appreciation of patients. Presented are 12 patients with avulsion of cervical spinal nerve roots that form either the brachial plexus upper trunk (n = 7), lower trunk (n = 1), or all three trunks (n = 4). The patients underwent the new bypass coaptation procedures with complete or partial return of motor and sensory function, which otherwise would be totally nonfunctional. The most dramatic results were noted in those patients who underwent operations within 6 weeks of injury. The results of these procedures offer patients a valid therapeutic modality for an enhanced quality of life after cervical nerve root avulsion. PMID- 8727146 TI - Atlantoaxial arthrodesis using Halifax interlaminar clamps reinforced by halo vest immobilization: a long-term follow-up experience. AB - Thirty-two patients who underwent atlantoaxial arthrodesis using Halifax interlaminar clamps and halo vests between January 1989 and December 1992 were reviewed. The atlantoaxial instabilities were related to trauma in 16 patients, including 14 patients with unstable odontoid fractures, 1 patient with a complex C2 fracture, and 1 patient with a disrupted transverse ligament. Of the other 16 patients, whose atlantoaxial instabilities were nontraumatic in origin, 9 had instabilities that were secondary to rheumatoid arthritis, 1 had instability that was secondary to tuberculous infection, and 6 had instabilities that were caused by os odontoideum. The patients were followed postoperatively with lateral cervical radiographs for an average of 37 months (range, 16-59 mo). Solid atlantoaxial arthrodeses were achieved in all (100%) of these 32 patients after 32 to 111 days (average, 84.5 d) of halo immobilization, indicating atlantoaxial arthrodeses can be reasonably anticipated when Halifax interlaminar clamps with autogenous iliac bone grafting are reinforced by halo vest immobilization for 3 months. PMID- 8727147 TI - Titanium aneurysm clips: Part I--Mechanical, radiological, and biocompatibility testing. AB - Most aneurysm clips are made of cobalt-based alloys. Although these clips are nonferromagnetic, they still produce artifact that degrades the quality of magnetic resonance (MR) images. A new aneurysm clip of pure titanium was developed to minimize artifact on postoperative MR images. We evaluated these clips in a series of mechanical tests in vitro, biocompatibility tests in rabbits, and radiological tests in greyhound dogs. The clip sizes and shapes matched those of conventional aneurysm clips. The average closing forces ranged between 151.6 and 181.8 g and were not diminished by repeated sterilization or stress. After > 20 million cycles of high-pressure and high-frequency pulsations, the clips did not open and the closing forces were not reduced. Titanium aneurysm clips implanted in the subarachnoid space of 12 rabbits for 1 or 6 months produced mild gliosis identical to that produced by implantation of cobalt alloy clips in 12 control rabbits. Based on pre- and postoperative weights and electron microscopic scans, the titanium implants did not corrode. The artifact on computed tomographic and MR imaging produced by a titanium clip placed on the internal carotid artery of a greyhound was less than that produced by an identical cobalt-chrome alloy clip by a factor of two to three. This study demonstrated that titanium aneurysm clips are mechanically equivalent to conventional clips, biocompatible, and corrosion resistant. Furthermore, titanium clips have superior imaging characteristics, creating less computed tomographic and MR imaging artifact and permitting better resolution of anatomic structures than cobalt alloy clips. PMID- 8727148 TI - Titanium aneurysm clips: Part II--Seizure and electroencephalographic studies in implanted rabbits. AB - Because titanium is widely used in neurosurgical procedures, we compared spontaneous and induced epileptiform activity in 12 rabbits with titanium clips implanted in the subarachnoid space with 12 rabbits with cobalt alloy clips and 6 rabbits that were not operated on that served as controls. Beginning 1 week after surgery, 30-minute electroencephalographic recordings were made at monthly intervals for 6 months. Recordings were scored by an electroencephalographer unaware of which treatment group was being recorded. In 48 recordings made during 6 months, no epileptiform activity was observed in any animal. Seizure threshold was evaluated by continuous intravenous injection of the convulsant drug, pentylenetetrazole (2 mg/kg/min), with continuous electroencephalographic recording. Time to spiking for the nonsurgical control group was 327 mean seconds +/- 181 standard deviation (SD), 216 mean seconds +/- 135 SD for the titanium group, and 389 mean seconds +/- 290 SD for the cobalt group. There were no significant differences among the groups (P = 0.17). Latency to behavioral tonicoclonic seizure was 1031 seconds +/- 537 SD for the group not operated on, 875 seconds +/- 334 SD for the titanium group, and 1267 seconds +/- 764 SD for the cobalt group. This study suggests that titanium clips are well tolerated within the brain and will not induce seizures. PMID- 8727149 TI - Titanium aneurysm clips: Part III--Clinical application in 16 patients with subarachnoid hemorrhage. AB - This report describes the first clinical use of newly developed titanium clips in the treatment of 16 patients with subarachnoid hemorrhage. There were no immediate or delayed complications related to the titanium clips. Thirteen patients had good outcomes, and one patient had moderate disabilities (mean follow-up, 5.4 mo). Two patients with Hunt and Hess Grade IV hemorrhages died postoperatively. The average cross-sectional areas of clip artifact on postoperative magnetic resonance imaging studies was 0.96, 1.36, and 1.05 cm2 on T1-, T2-, and intermediate-weighted images, respectively. In comparison, a matched control group with cobalt alloy clips had average cross-sectional areas of 3.13, 3.70, and 2.81 cm2 on T1-, T2-, and intermediate-weighted images, respectively. The average artifact volumes on gradient echo magnetic resonance images for titanium and cobalt alloy clips were 1.8 and 10.1 cm3, respectively. In addition, the gap on magnetic resonance imaging angiograms from clip artifacts was 0.9 cm with titanium and 2.6 cm with cobalt alloy clips. In conclusion, titanium aneurysm clips seem to be safe and effective and seem to reduce clip artifacts on magnetic resonance imaging threefold, compared with commercially available cobalt alloy clips. Because of this important advantage over conventional clips, titanium clips should be considered for routine use in aneurysm surgery. PMID- 8727150 TI - Normal and abnormal calcium homeostasis in neurons: a basis for the pathophysiology of traumatic and ischemic central nervous system injury. AB - Clinical recovery after central nervous system (CNS) trauma or ischemia may be limited by a neural injury process that is triggered and perpetuated at the cellular level, rather than by a lesion amenable to surgical repair. It is widely thought that one such process, a fundamental pathological mechanism initiated by CNS injury, is a disruption of cellular Ca2+ homeostasis. Because of the critical role of Ca2+ ions in regulating innumerable cellular functions, this major homeostatic disturbance is thought to trigger neuronal and axonal degeneration and produce clinical disability. We review those aspects of normal and pathological Ca2+ homeostasis in neurons that relate to neurodegeneration and to the application of neuroprotective strategies for the treatment of CNS injury. In particular, we examine the contribution of Ca(2+)-permeable ionic channels, Ca2+ pumps, intracellular Ca2+ stores, intracellular Ca2+ buffering systems, and the roles of secondary, Ca(2+)-dependent processes in neurodegeneration. A number of hypotheses linking Ca2+ ions and Ca2+ permeable channels to neurotoxicity are discussed with an emphasis on strategies for lessening Ca(2+)-related damage. A number of these strategies may have a future role in the treatment of traumatic and ischemic CNS injury. PMID- 8727151 TI - Clonal analysis of meningiomas. AB - Meningiomas are primary brain tumors arising from meningothelial cells. They usually grow slowly and are surgically easy to separate from the brain. A recent clonal analysis of meningiomas, using methylation-sensitive restriction fragment length polymorphisms, suggested a monoclonal origin. Using the same technique but with a highly informative X chromosome probe (M27 beta), we found that 17 (85%) of the 20 meningiomas analyzed were informative. Of the 17 informative tumors, 8 (47%) were monoclonal, 3 (18%) had loss of heterozygosity on the X chromosome, and, unexpectedly, 6 (35%) had a polyclonal pattern. Samples from two areas of one tumor showed a monoclonal pattern and loss of heterozygosity, respectively, on the X chromosome. A review of the histopathological and radiological features of the 17 informative tumors did not help to distinguish the clonal from the polyclonal tumors. We conclude that meningiomas are heterogeneous in clonal composition. PMID- 8727152 TI - Mild hypothermia, hypertension, and mannitol are protective against infarction during experimental intracranial temporary vessel occlusion. AB - A rabbit model of focal temporary ischemia was used to test the protection provided by mild hypothermia, hypertension, mannitol and the combination of the three methods. Twenty-four New Zealand White rabbits were divided into five groups as follows: a control group, a hypertension group (mean arterial blood pressure increased by 42 mm Hg), a hypothermic group (rectal temperature decreased by 6 degrees C), a mannitol group (1 g/kg of body weight, administered intravenously), and the triple-therapy group. The intracranial internal carotid artery, the middle cerebral artery, and the anterior cerebral artery were clipped for 2 hours and then underwent 4 hours of reperfusion. Blood pressure, rectal and brain temperature, blood glucose level, hematocrit, and arterial blood gases were monitored during the experiment. For measuring the infarction size, the brain was divided into 4-mm slices and stained with 2,3,5-triphenyltetrazolium chloride. The severity of the neuronal damage was also evaluated by conventional histological examination with hematoxylin and eosin staining. The infarct volume was 193.2 +/- 34.8 (standard error of the mean) mm3 for the control group, 32.3 +/- 22.6 mm3 for the hypertension group (P < 0.0005 versus control), 40.9 +/- 17.6 mm3 for the hypothermia group (P < 0.0005), 58.0 +/- 41.0 mm3 for the mannitol group (P < 0.005), and 0.9 +/- 0.9 mm3 for the triple-therapy group (P < 0.0001). The infarct volume of the triple-therapy group was smaller than that of the hypertension, hypothermia, and mannitol groups but the difference was not statistically significant. The combination of hypertension, mild hypothermia, and mannitol to protect against temporary focal ischemia provides a set of manipulations that is readily available for neurovascular procedures. PMID- 8727153 TI - Selective hypothermic perfusion of canine brain. AB - A method for selective brain cooling by profound hemodilution with cold Ringer's lactate solution was previously reported in 1992. We recently modified this technique by combining it with an ultrafiltration and rewarming circuit between the left jugular vein and the inferior vena cava. We used 12 beagle dogs to study the efficacy of selective cerebral hypothermia induced by this modified technique. The brain temperature decreased to 28 degrees C within 5.4 +/- 2.7 minutes and to 20 degrees C within 15.5 +/- 9.4 minutes. The lowest brain and rectal temperatures were 17.0 +/- 1.8 degrees C and 32.1 +/- 2.2 degrees C, respectively. All animals survived in good condition without evidence of neurological deficits until they were killed at 10 weeks. Histological examination of the brains with 2,3,5-triphenyltetrazolim chloride demonstrated no evidence of ischemic lesions, and even in the hippocampus, there was no evidence of ischemic neuronal damage. PMID- 8727154 TI - Mild hypothermia reduces penumbral glutamate levels in the rat permanent focal cerebral ischemia model. AB - Although the cerebroprotective effects of hypothermia in focal models of ischemia are undisputed, the underlying mechanisms of this protection are still subject to much controversy. To analyze whether mild hypothermia attenuates glutamate levels in the penumbra surrounding permanent focal infarcts, extracellular glutamate concentration was analyzed bilaterally by microdialysis 20 minutes before to 120 minutes after a middle cerebral artery occlusion (MCAO) in rats. Normothermic animals (n = 11) had a baseline glutamate concentration of 1.14 +/- 0.40 mumol/ml (standard error of the mean) before the MCAO. Extracellular glutamate levels increased gradually after vessel occlusion to peak at 10.1 +/- 1.45 mumol/ml 80 minutes after the MCAO. This level gradually decreased to 5.72 +/- 1.67 mumol/ml by 120 minutes. Hypothermic animals (n = 11) had a baseline glutamate concentration of 1.73 +/- 0.83 mumol/ml before the MCAO. Extracellular glutamate levels increased after vessel occlusion but stabilized at 3.47 +/- 1.37 mumol/ml 30 minutes after the MCAO and remained stable until completion of the experiment. There were no significant differences in cortical blood flow between the normothermic and hypothermic groups at any time during the experiment. Infarct volumes, expressed as a percentage of the volume of the right (ipsilateral) hemisphere, were 19.8 +/- 2.16% in the normothermic group and 13.0 +/- 1.42% in the hypothermic group (P < 0.02). Although the normothermic penumbral glutamate levels began to increase immediately after the MCAO, they did not peak until 80 minutes after occlusion. In contrast, the normothermic core glutamate levels peaked within 30 minutes after the MCAO. Glutamate diffusion from the core region to the penumbra might account for this delay. Hypothermic cerebroprotection might involve a reduction in the pool of potentially diffusable glutamate in the core region but have little direct effect on glutamate release in the penumbra. PMID- 8727155 TI - Harvey Cushing and Claude Beck: a surgical legacy. AB - Harvey cushing's contribution to neurosurgery and other surgical specialties has been profound. The accomplishments of his many pupils have further immortalized Cushing's role as a pioneering advocate of surgical science. Claude S. Beck, the first professor of cardiovascular surgery in the United States, was one such student. Beck's career and successes are illustrative of Cushing's guidance and enduring support for his surgical heirs. Beck's first encounter with Cushing evoked a spirit of fraternity and fashioned a respect and loyalty to Cushing that lasted for the duration of Beck's career. Cushing's personality and the methods by which he perpetuated a rich surgical tradition are illuminated by details of Beck's stay at the Peter Bent Brigham Hospital as a Cabot Fellow in Cushing's laboratory, by diary accounts of his later 6-week visit in 1927, and by the numerous telling correspondences between the two men. This article traces Beck's association with Cushing and provides unique insights into Cushing, his Cleveland connections, and his neurosurgical service at the Brigham. PMID- 8727156 TI - A history of the Neurological Institute of New York and its Department of Neurological Surgery. AB - The neurological institute of New York was founded in 1909 as the first hospital in North America devoted exclusively to the care of patients afflicted with neurological diseases. The Institute amalgamated with Columbia University's College of Physicians and Surgeons and The Presbyterian Hospital in New York City in 1928. The Department of Neurological Surgery developed under the successive leadership of Charles Elsberg, Byron Stookey, J. Lawrence Pool, Lester Mount, Edward Schlesinger, and Bennett Stein, each of whom brought unique qualities to the role of Department Chairman. This article traces the history of the Institute and its affiliates, present activities, and future plans. PMID- 8727157 TI - Progressive intracranial aneurysmal disease in a child with progressive hemifacial atrophy (Parry-Romberg disease): case report. AB - Intracranial aneurysms are uncommon in children, and their presence often leads to suspicion of a systemic connective tissue disorder. We describe the case of a young male patient with progressive hemifacial atrophy (Parry-Romberg disease) and multiple intracranial aneurysms, a previously undescribed association, and propose that a neural crest defect may be the underlying abnormality in this patient. At age 5 years, the patient was treated for a giant aneurysm of the left cavernous carotid artery with carotid ligation in the neck and a superficial temporal artery-middle cerebral artery bypass. At age 12 years, the patient was similarly treated for a giant aneurysm of the right cavernous carotid artery, which had progressed from a previously noted minute dilatation at age 5 years, with carotid ligation and a superficial temporal artery-middle cerebral artery bypass. At age 21 years, the patient was endovascularly treated for a de novo saccular aneurysm of the left posterior cerebral artery at the P1-P2 junction and a fusiform aneurysm of the distal left posterior cerebral artery. Various studies have suggested that the facial dermis, the subcutaneous tissues, and the skeleton, as well as the tunica media of the cervicocephalic arteries, all arise from neural crest cells, and a disorder of neural crest migration might explain the constellation of findings in this patient. PMID- 8727158 TI - Multicentric pleomorphic xanthoastrocytomas: case report. AB - We report the case of a 22-year-old woman who developed multicentric pleomorphic xanthoastrocytomas (PXAs) in the cerebral hemisphere. She underwent a first operation for a PXA in the right parietal lobe at the age of 7 years and a second operation at the age of 15 years in the right frontal lobe, remote from the previous tumor site. At age 22 years, she was found to have a tumor, which was a newly formed PXA, in the left occipital lobe. There was no recurrent tumor in the right frontal lobe. Clinical and pathological aspects of multicentric PXAs are reviewed and discussed. PMID- 8727159 TI - FasTRACKER-18. PMID- 8727160 TI - New jet irrigation bipolar system. AB - Irrigation bipolar systems are particularly useful for the washout of hematomas in subarachnoid hemorrhage during the acute stage of surgery and for preventing adherence of tissue to the bipolar tips during coagulation. These systems have become essential tools, and numerous improvements have been made to them. First, regarding the flow volume, both the interval of droplets and the size of individual water droplets can be adjusted. Second, this system is equipped with two major functions. The first is a preirrigation function, in which a small amount of irrigation water flows before operation of the bipolar coagulation. This wets the bipolar tips, eliminating any heat remaining from the previous use. Another function is postirrigation; after the coagulation has stopped, continuous irrigation is quickly provided to achieve a standby state by cooling down the tissue and the tips of the bipolar forceps. By using both of these functions, mild coagulation without adherence of burned tissue and clots to the bipolar forceps and minimization of residual heat during frequent use can be achieved. This article describes the new jet irrigation bipolar system and the use of an actual system that combines jet irrigation functions with the new cooling functions. PMID- 8727162 TI - Microsurgical carpal tunnel release. PMID- 8727161 TI - Ernst von Bergmann performs a brain operation by Franz Skarbina. PMID- 8727163 TI - Ventroposterior medial pallidotomy in patients with advanced Parkinson's disease. PMID- 8727164 TI - Genetic heterogeneity of inherited cerebral cavernous malformation. AB - OBJECTIVE: Cerebral cavernous malformation (CCM) is frequently an inherited disorder showing autosomal dominant transmission. Genetic analysis has localized a gene causing CCM to a segment of the long arm of human chromosome 7 (7q). This evidence derives from investigation of a small number of families, mostly of Hispanic American descent. In this study, we have tested whether inherited CCM is always due to mutation in this 7q gene, or whether mutation in other genes can cause CCM. METHODS: We have studied subjects from two non-Hispanic families with inherited CCM. The clinical features of CCM in these families are indistinguishable from those in kindreds in which CCM is due to mutation in the 7q gene. To test whether CCM in these kindreds is caused by a mutation on 7q, we compared the inheritance of CCM to the inheritance of genetic markers on 7q. RESULTS: Genetic analysis demonstrates independent inheritance of CCM and markers on 7q in both families studied. This evidence excludes mutation in the 7q gene as the cause of CCM in these families, with odds against CCM being due to mutation in 7q in each family of more than 100,000:1 and 100:1, respectively. CONCLUSION: These findings demonstrate that inherited CCM is not always caused by a mutant gene on 7q, indicating the presence of at least a second gene in which mutation can cause CCM. These results have implications for genetic testing and the pathogenesis of this disorder. PMID- 8727165 TI - Complementary therapeutic alternatives: what can we learn? PMID- 8727166 TI - Complementary medicine use in a dementia clinic population. AB - Complementary medicine is frequently utilized for a variety of chronic health problems. We evaluated its use among patients attending a Canadian dementia clinic. Using a telephone survey, we inquired about the use of alternative therapy, including nutritional supplements, herbal remedies, and chelation therapy, for problems with cognition. Only 9.6% of our patient population used complementary medicine as a treatment for cognitive problems. A further 29% used complementary medicines for general health promotion. While higher use might have been anticipated because of the limited conventional therapies available, we did not find a high prevalence of consumption of alternative medicine for cognitive problems. Knowledge of the use of these therapies is still important and should not be neglected. A nonjudgmental inquiry into all therapies being used (for whatever reason) should be part of the assessment of any patient with suspected dementia. PMID- 8727167 TI - Impaired awareness of deficits in Alzheimer disease. AB - This study examined the relation between awareness of memory and functional decline and cognitive function in patients with Alzheimer disease (AD). Twenty six patients with early AD and 16 nondemented elderly controls were studied. Awareness of deficits was determined by using (a) a discrepancy score between subject's and caregiver's ratings on a memory questionnaire, (b) a discrepancy score between subject's and caregiver's ratings on an activities-of-daily-living scale, and (c) a clinical rating of dementia awareness for patients. Whereas self ratings of memory and activities of daily living were not significantly different between AD patients and controls, these two measures differed significantly when AD patients' ratings were compared with those of their caregivers. Measures of awareness of deficits correlated with one another and were primarily associated with performance on tests of executive and visuospatial functions but not with depression. Early AD is characterized by a failure of self-monitoring. Deficits in self-monitoring have been proposed to occur after damage to the frontal lobes and other cerebral areas. Impaired awareness of memory and functional deficits in AD is related to cognitive impairments, which may involve frontal and right hemisphere connections. PMID- 8727168 TI - Reductions in membrane proteins and lipids in basal ganglia of classic Alzheimer disease patients. AB - Gangliosides and major membrane components were determined in caudate nucleus, putamen, and hippocampus of 12 cases with Alzheimer disease (AD) type I, also termed presenile or pure AD, and in age-matched controls. The concentration of gangliosides, a marker for axodendritic arborization, was reduced to 71% in caudate nucleus, 82% in putamen, and 66% in hippocampus of that in the controls. Significant diminution of total dry solids, protein, and phospholipids was also found in caudate nucleus and was most pronounced in hippocampus. The early signs of extrapyramidal features have been emphasized in AD type I. We now provide evidence that the neostriatum is affected in AD type I (the putamen, however, to a lesser extent than the caudate nucleus). The biochemical changes in these nuclei can be significantly related to scores of impairment of motor performance. PMID- 8727169 TI - Physical aggression and associated factors in probable Alzheimer disease. AB - Aggressive behavior is thought to be pervasive among and devastating to demented patients and their caregivers. This study investigated the prevalence of physical aggression in demented inpatients with Alzheimer disease (AD). Additionally, the characteristics and psychiatric symptoms that may be associated with aggression were tested. Forty-seven patients with probable AD were included in this study. Physical aggression was reported in 27 (57.4%) of the patients. These patients were older, had a later onset of dementia, and had a higher prevalence of misidentification, activity disturbances, and day/night disturbance. The relationship between these factors and physical aggression needs further clarification, which may help provide effective predictors and treatment of the aggressive behavior. PMID- 8727170 TI - Sexual behavioral changes in Alzheimer disease. AB - The frequency and correlates of sexual behavioral changes in Alzheimer disease (AD) were assessed in two studies. In the first study, we reviewed the medical records of 135 consecutive community-living patients who fit the criteria of the National Institute of Neurological and Communicative Disorders and Stroke/Alzheimer Disease and Related Disorders Association for probable or possible AD, and we asked spouses to complete a questionnaire that included two questions about sexual activity. Indifference to sexual activity was reported by 70% of the spouses, and sexual behavioral modifications were reported by 50%. No correlation was found between sexual changes and the general characteristics of the patients, the severity of the disease, or depressive symptomatology. Sexual changes were correlated to the severity of behavioral and mood disorders, mainly to a reduction of activity and emotional deficit. Seventy-seven of the patients had a second examination after an interval of 18.9 +/- 9.9 months. The links between sexual changes and the other variables were similar to those found by the first examination. In the second study, a questionnaire was sent by mail to the spouses of 100 patients. No relationship was found between sexual changes and the severity of cognitive deficits, previous sexual agreement, age, or gender. Sexual changes were considered a factor of maladjustment for the couple by 46% of the spouses. PMID- 8727171 TI - A treatment and withdrawal trial of besipirdine in Alzheimer disease. AB - Besipirdine hydrochloride (HP 749) is an indole-substituted analog of 4 aminopyridine. Besipirdine enhances both cholinergic and adrenergic neurotransmission in the central nervous system. The present study examined the efficacy and tolerability of two doses of besipirdine (5 and 20 mg b.i.d.) in 275 patients with Alzheimer disease during 3 months of treatment and for 3 months after withdrawal of treatment. Assessment after withdrawal of treatment was used in an effort to distinguish persistent efficacy attributable to a neuroprotective mechanism from reversible symptomatic efficacy. Besipirdine was generally well tolerated. The level of performance on the cognitive subscale of the Alzheimer Disease Assessment Scale (ADAS-Cog) was sustained during 3 months of treatment with besipirdine, whereas some deterioration in the performance of patients treated with placebo was observed over the same period. The small difference between active and placebo treatment groups approached, but did not reach statistical significance in the primary intent-to-treat analysis (p = 0.067); analysis of patients who completed all assessments was supportive (p = 0.031). Global ratings using the Clinician Interview-Based Impression of Change did not detect a besipirdine treatment benefit, possibly because of an adverse effect on mood and behavior in some patients. A high ratio of adrenergic to cholinergic potency may have resulted in the adverse effects of besipirdine and hence its failure to support the hypothesis that multiple neurotransmitter treatment may be more efficacious than monotherapy. The efficacy apparent on the ADAS-Cog after 3 months of treatment did not persist 3 months after withdrawal of treatment, suggesting that the benefit was symptomatic. This study provides a practical example of the use of treatment withdrawal assessment to distinguish neuroprotective from symptomatic efficacy. PMID- 8727172 TI - Extrapyramidal motor signs in clinically diagnosed Alzheimer disease. AB - We reviewed clinical case series published over a 10-year period addressing the cross-sectional frequency, incidence, and diagnostic and prognostic significance of extrapyramidal signs (EPS) in Alzheimer disease (AD). The review was prompted by recent reports of Lewy body (LB) pathology in the brains of many AD patients and the association of LB pathology with clinical parkinsonism in AD. In the clinical case series reviewed, we evaluated several possible determinants of prevalent EPS, including neuroleptic use, EPS assessment technique, and dementia severity. Neuroleptics were a well recognized cause of parkinsonism in these reports, though some failed to document the frequency of neuroleptic use. Assessment methods were also important: Studies using structured clinical research scales to rate EPS reported higher frequencies than studies employing routine neurological examination. The relationship between parkinsonism and dementia severity was complex. Some studies found bradykinesia, facial masking, and parkinsonian postural changes even in mildly demented, neuroleptic-naive AD patients. Rigidity, on the other hand, became increasingly common as dementia progressed. AD patients with EPS showed faster cognitive and functional decline and earlier death than those without EPS, even after consideration of differences in initial dementia severity. In the differential diagnosis of dementia with parkinsonism, LB disease in its various forms, including AD with LB, is the principal diagnostic consideration. Future studies of parkinsonism in AD should employ standardized clinical rating scales and should exclude patients on neuroleptics or analyze their results separately. Investigators should report frequencies for individual parkinsonian signs in addition to the overall prevalence of EPS to facilitate meaningful comparisons across studies. PMID- 8727173 TI - Relations of the superficial musculoaponeurotic system to the orbit and characterization of the orbitomalar ligament. AB - The orbital and eyelid relationships of the superficial musculoaponeurotic system (SMAS) were studied in human cadavers. Using gross and microscopic techniques, the SMAS was found to be intimately related to the eyelids and orbicularis oculi muscle, and to have distinct orbital bony attachments. Sub-SMAS fat in the malar region was found to be continuous with the submuscular fat in the eyebrow region. The malar sub-SMAS fat continued superiorly into the lower eyelid above the inferior orbital rim, as a postorbicularis layer. A bony attachment emanating from the inferior orbital rim, the orbitomalar ligament, traveled through the orbicularis oculi muscle in a lamellar fashion prior to inserting into the dermis. The cutaneous insertion of this attachment corresponds to the malar and nasojugal skin folds. With aging, relaxation of the orbitomalar ligament allows inferior migration of orbital fat, in addition to the anterior migration that occurs through an attenuated orbital septum. These findings have implications not only in cosmetic surgery but also in the understanding of facial soft tissue changes that occur with aging. PMID- 8727174 TI - Cyanoacrylate tissue adhesive in blepharoplasty. AB - This article reports the results of an animal study designed to evaluate the usefulness of butyl 2-cyanoacrylate (Histoacryl) in blepharoplasty skin closure. Bilateral upper and lower lid blepharoplasty skin incisions were made on seven pigs. We compared the wounds closed with 6.0 nylon to those closed with Histoacryl at 1, 2, 4, and 9 weeks postoperatively. The tensile strength of the "glued" incisions was significantly greater than that of the sutured incisions only at the 9-week postoperative interval (p < 0.05). There was no significant difference in bond strength, however, between the two methods at 1, 2 and 4 weeks postoperatively. Histopathologic analyses of sutured and glued wounds were compared at the same designated postoperative periods. No deleterious effects were found in the overall healing of the cutaneous wounds closed with Histoacryl. PMID- 8727175 TI - Involutional entropion repair by posterior lamella tightening and myectomy. AB - Involutional entropion is a common eyelid malposition of diverse etiology that may recur after surgical repair. Laxity of the tarsoligamentous complex combined with posteriorly directed rotational force exerted by the orbicularis, in our view, seems to be the most important in the production of entropion. A surgical technique has been developed that is carried out through a standard transcutaneous lower eyelid blepharoplasty incision. It includes tarsoligamentous tightening at the lateral canthus, bolstering of the lateral canthal tendon, and partial orbicularis myectomy. Forty-two procedures in 35 consecutive patients (29% for recurrent entropion) have been performed and evaluated (mean follow-up, 33 months). There have been no recurrences, and the esthetic outcome has been very good. PMID- 8727176 TI - Congenital cystic eye. AB - We present a rare case of congenital cystic eye associated with holoprosencephaly and tetralogy of Fallot. The orbital cyst was diagnosed preoperatively by computed tomography (CT) scan, and we present the first magnetic resonance imaging (MRI) finding in such a disorder. The orbital cyst expanded over 7 months of follow-up. The cyst was excised in toto and studied by immunohistochemistry, and its content was analyzed biochemically. The patient achieved an acceptable cosmetic result. PMID- 8727177 TI - Use of demineralized bone implants in orbital and craniofacial reconstruction and a review of the literature. AB - A retrospective study and review of the literature was performed on the use of demineralized bone implants for the correction of orbital and craniofacial defects. Demineralized bone implants heal by endochondral osteogenesis, inducing a transformation of local cells, as well as by osteoconduction, similar to autogenous grafts. They induce the chemotaxis and transformation of mesenchymal cells into chondroblasts, followed by ossification. They also act as a scaffold, with bone resorption taking place simultaneous with bone formation. This study reviewed 21 patients and 31 orbits, in which demineralized bone was used for orbitocranial reconstruction for congenital deformities (nine patients), after surgery for orbital fractures (seven patients: four floor, three roof), and orbital tumors (five patients). The surgical technique is described, and the results are discussed. The follow-up period averaged 15 months (6 months to 33 months). The overall resorption rate of the demineralized bone implants was estimated based on follow-up radiologic studies (facial films, computed tomography scans, and magnetic resonance imaging scans), as well as clinical examinations. All patients had a satisfactory to excellent result. The demineralized bone and Grafton (Osteotech, Shrewsbury, NJ, U.S.A.) was obtained from the Musculoskeletal Transplant Foundation of Holmdel, New Jersey, and it was all processed and reconstituted in a standard manner with Alloprep System. Surgical complications were cerebrospinal fluid leaks (one patient) with infection, transient chemosis, enophthalmos, and hypophthalmos. There were no complications related to demineralized bone alone. Properly prepared demineralized bone is a safe material for orbital reconstruction that eliminates the need for a second operative site to harvest a bone graft. PMID- 8727178 TI - Adverse effects of bone wax in surgery of the orbit. AB - The goal of this article is to establish the incidence and scope of adverse reactions to bone wax in a large orbital surgical series. We report two patients with bone wax granulomas of the orbit as a remote surgical complication. These are the first reported cases of adverse reactions to bone wax in the ophthalmic literature. A chart review was conducted on all patients from the University of British Columbia Orbit Clinic that had surgery with temporary or permanent removal of orbital bone. Two patients with bone wax granulomas were identified. In one case, intraoperative cultures grew Staphylococcus aureus, confirming that the wax may indeed act as a nidus for infection. No cases of pseudoarthrosis have occurred. This syndrome of chronic granulomatous giant cell foreign body inflammation has characteristic clinical, radiologic, and histopathologic features. The literature regarding adverse reactions to bone wax is reviewed, and specific implications for orbital surgery are discussed. PMID- 8727179 TI - Magnetic resonance imaging versus bone scan for assessment of vascularization of the hydroxyapatite orbital implant. AB - We prospectively studied 10 patients who were status postenucleation and primary placement of the hydroxyapatite orbital implant. Both the technetium-99m bone scan and gadolinium-enhanced magnetic resonance images (MRI) were obtained on the same day at variable time points in the postoperative period in order to assess the degree of vascularization. Up to 78% of the bone scans were interpreted as being completely vascularized, while only 10% of the corresponding MRI scans were consistent with complete vascularization. Cost analysis showed that MRI was cost effective imaging modality. We conclude that contrast-enhanced MRI provides a more accurate assessment of vascularization of the hydroxyapatite orbital implant when compared to bone scan. PMID- 8727180 TI - Draining cutaneous fistula associated with infection of hydroxyapatite orbital implant. AB - The implantation of an orbital hydroxyapatite implant was complicated by conjunctival dehiscence, cutaneous fistula formation, and infection with Staphylococcus aureus. Pathologic examination of the sphere 2 years after its implantation revealed reduction in the size of the implant, peripheral lamellar bone formation and central necrosis. This is the first report of this constellation of complications with hydroxyapatite spheres positioned in the orbit. The 2-year interval between implantation and removal of the sphere is the longest reported in a case with histopathologic analysis. PMID- 8727181 TI - Oculoplastic pearls. PMID- 8727182 TI - Binocular indirect ophthalmoscopy using a standard fiberoptic surgical headlight. AB - Binocular indirect ophthalmoscopy may be performed in the operating room with a fiberoptic surgical headlight and condensing lens, eliminating the need for the indirect ophthalmoscope headpiece. The resultant aerial inverted stereo image has enhanced stereopsis but is otherwise identical to that obtained with the headpiece. Safe power settings for individual headlights can be determined with a photometer by measuring illuminance from a standard indirect ophthalmoscope and comparing this value to the intensities obtained at each voltage setting of the headlight. Illuminance from four different headlights was measured, and settings that yielded light output equivalent to the average indirect ophthalmoscope were designated for each headpiece. The maximum indirect ophthalmoscope illuminance values were averaged and found to be 2.0 watts/m2 (W/m2) for the 20-diopter (D) lens and 1.8 W/m2 for the 30-D lens. All of the headlights could be used at transformer settings that produced light intensities at or below the average maximum indirect ophthalmoscope illuminance. A stereoscopic image was obtained with each of the headlights, with both the 20-D and 30-D lenses. We conclude that standard fiberoptic surgical headlights may be used for stereoscopic fundus examination. The average maximal light intensity of fiberoptic surgical headlights is greater than that of the average indirect ophthalmoscope. However, each fiberoptic headlight can be operated at brightnesses equal to or below that of the maximum intensity of the indirect ophthalmoscope, and therefore, retinal toxicity is unlikely for brief retinal evaluations. PMID- 8727183 TI - Granuloma annulare of the eyelid. AB - A superficial (dermal) granuloma annulare (GA) of the eyelid developed in a 69 year-old woman who initially had no evidence of precipitating causes, including trauma, tuberculosis, octopus bite, lupus vulgaris, actinic damage, sarcoidosis, diabetes mellitus, rheumatoid arthritis, systemic lupus erythematosus, or rheumatic fever. She later developed complete bilateral nasolacrimal duct obstruction that led to dacryocystorhinostomy on the right side. Systemic workup showed evidence of a lupus-like syndrome. Although deep, subcutaneous GAs have been reported in the periocular tissues, episclera, and orbit in children and young adults, a superficial dermal GA of the eyelid in an elderly patient is distinctly rare. The characteristic histopathologic feature of both superficial and deep GAs is a necrobiotic granuloma in which necrotic collagen is surrounded by a zone of histiocytes and fibroblasts. This case demonstrates that superficial GA of the eyelid may be associated with an underlying lupus-like syndrome. This case also raises the question of whether GA of the eyelid and lupus erythematosus may be associated with bilateral nasolacrimal duct obstruction. PMID- 8727184 TI - The stress gene response in brain. AB - Changes in gene expression in the brain in response to adverse conditions, such as ischemia or excitotoxin exposure, may be part of the injury process or represent an adaptive response which may be protective during subsequent stressful events. In this review we have considered the regulation, functions and potential relationships to the pathophysiology of ischemia of several major groups of stress-induced genes, including those of the M(r) 27,000, 32,000 (heme oxygenase), 70,000 and 90,000 heat shock protein families, the glucose-regulated proteins, glucose transporters and ubiquitin. Patterns of gene expression in several injury models, including focal and global ischemia, excitotoxin/ seizure related injury and hyperthermia are reviewed. In vitro expression studies and the phenomenon of ischemic tolerance are also discussed. It is concluded that stress gene expression provides a useful marker of cellular injury, and that disjunction of mRNA and protein expression may be indicative of imminent death in cells which survive the initial insult. Though other stress proteins may play a role, it seems unlikely that neuronal hsp70 expression is a major contributor to ischemic tolerance. PMID- 8727185 TI - Brain damage due to cerebral hypoxia/ischemia in the neonate: pathology and pharmacological modification. AB - Brain damage due to an episode of cerebral hypoxia/ischemia remains a major problem in the human infant, providing impetus for the testing of potential neuroprotective agents in animal models. Although these animal models do not mirror the human pathology exactly (e.g., with respect to regions vulnerable to damage), they usually have the histological characteristics of gray matter hypoxic/ischemic injury in the human. An important factor in comparing models directly is the stage of development of the brain at birth, which varies widely between species. Approaches to prevent or treat cerebral hypoxic/ischemic damage in neonates have paralleled those in adults. However, most of these results should be interpreted cautiously, since neonatal rat models with little concurrent physiological monitoring are often used. As in adults, moderate hypothermia during the insult or a preconditioning stress prior to the insult has prevented hypoxic/ ischemic brain damage. Different from adults is the demonstration that pretreatment with moderate doses of glucocorticoids or hyperglycemia during the hypoxic/ ischemic insult protects the brain against infarction. Partial protection, primarily in neonatal rats, has also been produced by pretreatment with voltage-sensitive calcium channel antagonists, free radical scavengers, growth factors, gangliosides, anticonvulsants, antiinflammatory agents, and nitric oxide synthase inhibitors. Posttreatment has been effective with a few agents. The most consistent has been the protective effect observed with glutamate receptor antagonists administered before but also up to 4 h after the insult. The effects of most of these therapies on blood glucose, body temperature, and/or the systemic circulation should be measured and the protective effects confirmed in larger species prior to considering clinical applications. PMID- 8727186 TI - An anatomic evaluation of L5 nerve stretch in spondylolisthesis reduction. AB - STUDY DESIGN: Lumbosacral spondylolisthesis was simulated using four embalmed human spines, and the path of the L5 nerve was studied. OBJECTIVES: To quantify the change in length of the L5 nerve root associated with reduction of spondylolisthesis, correction of slip angle, and changing disc height. SUMMARY OF BACKGROUND DATA: Stretch injury to the lumbar nerves remains a complication of spondylolisthesis reduction. To date, no anatomic studies have been performed to quantify this effect of reduction on the lumbar nerves. METHODS: The L5 vertebral body and the sacrum of four embalmed human spines were constrained in an adjustable jig, and the length of a simulated nerve was determined for various position variables--sagittal translation (0-100% slip), slip angle (-40 degrees to +20 degrees), and disc height (5 or 10 mm). Two standard points of reference were chosen to represent fixed points along the path of the L5 nerve. An inelastic cord was used to measure the path length between these points as L5 was reduced from 100% to 0% slip. Testing was performed using a 5-mm and a 10-mm disc height. The effect of varying slip angle alone was also studied. RESULTS: The effect of spondylolisthesis reduction and slip angle correction on nerve length varied depending on the location of L5 with respect to the sacrum. There was an increasing effect of partial reduction on nerve length as L5 approached full reduction. Initially, little strain was produced in the L5 nerve as L5 was reduced in higher grade slips. However, as L5 approached full reduction, the strain per increment of reduction increased rapidly. On average, the mean nerve strain was 4.0% for the first 50% of reduction and 10.0% for the second half of reduction. Increasing lordosis relaxed the nerve in high-grade slips and stretched the nerve in fully reduced slips. At 100% slip, the mean nerve excursion decreased 5.1 mm (nerve slackening) when L5 was rotated from +20 degrees to -40 degrees. At 0% slip, the mean nerve excursion increased 3.1 mm (nerve stretch). Increasing disc height directly stretched the L5 nerve. However, given a larger disc height, the strain on the nerve per increment of reduction was less than for the smaller height. CONCLUSION: The findings suggest that the risk of stretch injury to the L5 nerve with reduction of a high-grade spondylolisthesis is not linear; with 71% of the total L5 nerve strain occurring during the second half of reduction, partial reduction may be a significantly safer treatment approach for high-grade spondylolisthesis than complete reduction. Correction of lumbosacral kyphosis in high-grade spondylolisthesis may be protective of the L5 nerve. PMID- 8727187 TI - The role of melatonin in the pathogenesis of adolescent idiopathic scoliosis. AB - STUDY DESIGN: A matched, case-control study comparing melatonin production in female patients with and without adolescent idiopathic scoliosis. OBJECTIVES: To determine whether melatonin production is decreased in adolescent idiopathic scoliosis. SUMMARY OF BACKGROUND DATA: A central etiology for idiopathic scoliosis has never been established. Previous authors have produced experimental scoliosis in chickens after pinealectomy, preventable by administration of melatonin. They suggested that a defect in melatonin synthesis might be involved in the pathogenesis of human idiopathic scoliosis. METHODS: Nine female adolescents with no medical problems, normal neurologic examinations, radiographic idiopathic scoliosis of 15-40 degrees, and Risser Stage I-III were in the patient group. Eighteen healthy adolescent girls with no medical problems, a negative school screening, and no family history of scoliosis were control subjects. Patients and control subjects were matched for age, weight, Tanner stage, sleep duration, and light exposure by multiple linear regression. Nighttime and daytime urine samples were analyzed for melatonin by high performance liquid chromatography. RESULTS: Although nighttime melatonin levels were significantly higher than daytime levels in all volunteers (P < 0.00002), there were no significant differences in nighttime (P > 0.63) or daytime (P > 0.78) melatonin levels between patients and control subjects, even after matching by multiple linear regression analysis. A statistical analysis demonstrated that if a melatonin deficiency of 25% or more did exist in patients with scoliosis compared with control subjects, the likelihood that it would have been detected in this study was more than 98%. CONCLUSION: Although melatonin deficiency may cause scoliosis in the chicken, this study suggests that it is not a mechanism in the pathogenesis of adolescent idiopathic scoliosis in humans. PMID- 8727188 TI - Melatonin. A possible role in pathogenesis of adolescent idiopathic scoliosis. AB - STUDY DESIGN: The serum melatonin levels during 24-hour periods were compared between patients with idiopathic and age-matched normal control subjects. OBJECTIVE: To find if the melatonin deficiency may have some role for progression or etiology of idiopathic scoliosis in humans. SUMMARY OF BACKGROUND DATA: Experimentally induced scoliosis in chicken by pinealectomy can be attributed to the defect in melatonin metabolism. METHOD: Blood samples were correlated every 3 hours during 24-hour periods, and serum melatonin levels were measured and statistically analyzed. RESULTS: The level of melatonin, integrated concentration through 24 hours and night time (0:00 am-6:00 am), in the patients who had progressive curve (more than 10 degrees of progression in the previous 12 months) was significantly lower than the level in the patients who had a stable curve (less than 10 degrees of progression in the previous 12 months) or in the control subjects (P < 0.05). CONCLUSION: The study suggests that normal melatonin synthesis or metabolism may have crucial role in regulating normal spine growth. The level of melatonin appears to be a useful predictor for progression of spine curvature in idiopathic scoliosis. PMID- 8727190 TI - Mechanical modulation of vertebral body growth. Implications for scoliosis progression. AB - STUDY DESIGN: The authors developed a rat-tail model to investigate the hypothesis that vertebral wedging during growth in progressive spinal deformities results from asymmetric loading in a "vicious cycle." OBJECTIVES: To document growth curves with axial compression or distraction applied to tail vertebrae to determine whether compression load slows growth and distraction accelerates it. SUMMARY OF BACKGROUND DATA: Progression of skeletal deformity during growth is believed to be governed by the Hueter-Volkmann law, but there is conflicting evidence to support this idea. METHODS: Twenty-eight 6-week-old Sprague-Dawley rats were assigned to one of three groups: compression loading, distraction loading, or sham (apparatus applied without loading). Under general anesthesia, two 0.7-mm diameter stainless steel percutaneous pins were used to transfix each of two vertebrae. The pins were glued to 25-mm diameter external ring fixators. Springs (load rate, 35 g/mm) were installed on three stainless steel threaded rods that were passed through holes in each ring and compressed with nuts to apply compression or distraction forces between 25-75% of bodyweight. Vertebral growth rates in microns/day were measured by digitizing the length of the vertebrae images in radiographs taken 0, 1, 3, 5, 7, and 9 weeks later. RESULTS: The loaded vertebrae grew at 68% of control rate for compressed vertebrae and at 114% for distracted vertebrae. (Differences statistically significant, P < 0.01 by analysis of variance.) For the compressed vertebrae, the pinned vertebrae, which were loaded at one of their two growth cartilages, grew at a reduced rate (85%), although this effect was not apparent for the distraction animals. CONCLUSIONS: The findings confirm that vertebral growth is modulated by loading, according to the Hueter-Volkmann principle. The quantification of this relationship will permit more rational design of conservative treatment of spinal deformity during the adolescent growth spurt. PMID- 8727189 TI - The human lumbar endplate. Evidence of changes in biosynthesis and denaturation of the extracellular matrix with growth, maturation, aging, and degeneration. AB - STUDY DESIGN: The authors measured concentrations of specific molecules reflecting matrix synthesis and degradation in 121 human endplates and correlated them with aging and grade of degeneration. OBJECTIVES: Abnormal endplate development has been implicated in many spinal abnormalities, yet little is known about endplate matrix component turnover. SUMMARY OF BACKGROUND DATA: Techniques are available to perform an in situ investigation of matrix component turnover with aging and degeneration. METHODS: Newly synthesized aggrecan and Type I and Type II procollagens were measured with recently developed immunoassays. Percentage of denatured Type II collagen was assessed with a new enzyme-linked immunosorbent inhibition assay. RESULTS: Synthesis in endplates, measured by content of an aggrecan marker (846) and content of Types I and II procollagen markers (CPI and CPII), is highest in the neonatal and 2- to 5-year age groups and steadily diminishes with increasing age. However, in the oldest age group and in highly degenerated discs, the CPI epitope level increased significantly. Percentage of denatured Type II collagen, assessed by the presence of an epitope exposed with the cleavage of Type II collagen, increased from the neonatal to the 2- to 5-year age groups. The percentage progressively decreased with increasing age. However, it significantly increased in endplates from highly degenerated discs. CONCLUSIONS: The authors identified three matrix turnover phases, related to age and grade of degeneration. Phase I (growth) is characterized by active synthesis of matrix molecules and active denaturation of Type II collagen. Phase II (aging and maturation) is distinguished by a drop in synthetic activity and a reduction in denaturation of Type II collagen. Phase III (degenerative) is illustrated by an increase in Type II collagen denaturation and Type I procollagen synthesis, both related to grade of tissue degeneration. PMID- 8727191 TI - Overpowering the crankshaft mechanism. The effect of posterior spinal fusion with and without stiff transpedicular fixation on anterior spinal column growth in immature canines. AB - STUDY DESIGN: This experimental study was designed to test the hypothesis that posterior spinal fusion and internal fixation, using a stiff transpedicular construct, would withstand additional anterior column growth without the need for an anterior procedure and would prevent the development of deformity secondary to asymmetric growth of the anterior column in the immature canine model. OBJECTIVES: This model revealed that a mechanical epiphysiodesis of the anterior spinal column can be created through a posterior approach in an immature growing animal using a stiff transpedicular construct. These results were correlated to the crankshaft mechanism clinically to provide a possible rationale for transpedicular posterior spinal instrumentation and fusion, without a concomitant anterior procedure, in immature patients. SUMMARY OF BACKGROUND DATA: All operative immature canines underwent posterior fusion of L1-L5 vertebral bodies with autogenous iliac crest bone graft. Instrumented canines underwent the additional placement of transpedicular screws at L1, L3, and L5 as well as 3/16 inch rods and a transverse connector. Previous studies have revealed that continued anterior spinal column growth after posterior arthrodesis causes a resultant deformity in quadrupeds. No previous study has assessed whether a transpedicular construct can overpower the anterior spinal growth plate in an immature growing model. METHODS: Twenty-five skeletally immature canines were randomized into four groups: control, fusion only, screw and fusion, and instrumentation and fusion. Disc space growth, vertebral body growth, and sagittal plane angulation were the variables analyzed. Management response variables were visualized graphically and radiographically. RESULTS: Posterior tethering and anterior column growth occurred in the noninstrumented fusion groups, producing substantial lordosis. Anterior column growth was arrested in the instrumented canines as demonstrated by decreased vertebral body length, disc space narrowing, and, most importantly, prevention of lordosis. CONCLUSIONS: The present study confirmed that in immature canines anterior column growth continues after posterior fusion without instrumentation. The magnitude of this growth, combined with a posterior tether, is sufficient to cause significant lordosis. The results are the first to document that a stiff posterior spinal instrumentation system is sufficient to overpower the residual anterior growth centers, even in the presence of a posterior tether (fusion mass). This technique creates a mechanical epiphysiodesis evidenced by arresting vertebral body length, narrowing disc space, and preventing lordosis, thus thwarting the deformity producing mechanism without an additional anterior procedure. PMID- 8727192 TI - Is the nucleus pulposus a solid or a fluid? Mechanical behaviors of the nucleus pulposus of the human intervertebral disc. AB - STUDY DESIGN: A new technique to measure the viscoelastic behavior of the nucleus pulposus in shear was used to assess its solid and fluid characteristics. OBJECTIVES: To review existing knowledge on mechanical behaviors of the nucleus pulposus, and to develop a new technique to study the viscoelastic behaviors of isolated nucleus pulposus samples in torsional (pure) shear under transient and dynamic conditions. SUMMARY OF BACKGROUND DATA: Numerous studies have investigated the swelling behavior of the nucleus and found the swelling pressure to range approximately 0.05-3 MPa, depending on loading conditions. Very few studies, however, have investigated the load-deformational behaviors of the nucleus pulposus. METHODS: Thirteen nondegenerate samples of nucleus pulposus were harvested from lumbar discs and tested in torsional shear under transient and dynamic test conditions. A linear viscoelastic law with variable amplitude relaxation and dynamic frequency sweep experiments. The coefficients of the viscoelastic law were determined from the stress relaxation experiments, whereas the dynamic shear modulus and phase shift angle were determined from the frequency sweep. RESULTS: The nucleus exhibits significant viscoelastic effects in shear. Under transient conditions, the stress relaxed to values near zero, which is indicative of the "fluid-like" behaviors of the nucleus. Under dynamic conditions, however, the material parameters for the nucleus, magnitude of the complex modulus (7-21 kPa), and phase angle (23-31 degrees) were more characteristic of a viscoelastic solid. The authors' proposed stress-strain law exhibited excellent agreement with the viscoelastic data. CONCLUSIONS: In response to shear deformations, the nucleus pulposus exhibited significant viscoelastic effects, characteristic of a fluid and a solid. Whether the nucleus pulposus behaves more as a fluid or a solid in vivo depends on the rate of loading. PMID- 8727193 TI - Effect of sacral and iliac instrumentation on strains in the pelvis. A biomechanical study. AB - STUDY DESIGN: This study is a biomechanical study on human cadaver pelves using strain gauges to measure strains at different locations in response to different load states. OBJECTIVES: To examine the changes in strains, at different sites on the pelvis, and after instrumentation of the lumbosacral joint and the lumbosacroiliac joint. SUMMARY OF BACKGROUND DATA: Pelvic girdle pain after long instrumented fusions to the sacrum has been described and studied along with sacroiliac and pubic instability. Fractures of the pelvic rami or the iliac wing have been reported in patients after surgical fusions that incorporated the sacrum and sacroiliac joint. METHODS: Pelves with lumbar spine attached had three axis rosette strain gauges cemented to specific sites on the pelves (ilium [correction of ileum] and rami). Specimens were tested in different configurations involving lumbosacral and sacroiliac instrumentation. Iliac bone harvest was also tested. Strain values were computed and compared for each configuration. RESULTS: The "Galveston" and "sacral" instrumentation configurations caused significant stress shielding in pelves under all compression loads applied. Axial rotations on the instrumented specimens caused higher strains at all sites. Harvesting of bone from the ilium did not alter the strains seen at the different sites. CONCLUSIONS: This study suggests significant stress shielding provided by sacro and sacroiliac instrumentation systems, supporting the possibility of development of device-related osteopenia that may predispose patients to late, insufficiency-type fractures as their activity increases. The rigid instrumentation of the lumbosacral spine was found to increase the stress during torsion, which may lead to failure during activity. PMID- 8727194 TI - Geometric analysis of coronal decompensation in idiopathic scoliosis. AB - STUDY DESIGN: Frontal plane geometry of postoperative curves was analyzed using a geometric model to investigate the relationship between coronal decompensation and postoperative apical shifts from the center sacral line for various thoracic and lumbar Cobb angles. OBJECTIVE: To determine if a balanced spinal configuration is possible when the postoperative lumbar curve is larger than the thoracic curve, and to determine the limits on the postoperative magnitude of the lumbar curve relative to the thoracic curve beyond which a spinal configuration with acceptable balance cannot be achieved. SUMMARY OF BACKGROUND DATA: Previous studies have suggested that overcorrection of the primary thoracic curve may be the principal cause of coronal decompensation after selective thoracic correction and fusion in King Type II curves. Also, other causative factors, such as inappropriate selection of fusion levels and hook patterns, have been implicated as possible reasons for decompensation after Cotrel-Dubousset instrumentation for idiopathic scoliosis. METHODS: Postoperative thoracic curves of 20 degrees, 25 degrees, and 30 degrees were simulated on a model spine. For each thoracic Cobb angle, three left lumbar curves were simulated with the lumbar curve larger than thoracic by 5 degrees, 10 degrees, and 15 degrees. For each combination of thoracic and lumbar Cobb angles, spinal configurations corresponding to different lateral shifts of the thoracic and lumbar apical vertebrae from the center sacral line were obtained. RESULTS: For a given combination of postoperative thoracic and lumbar Cobb angles, there is an optimal range of postoperative lateral distance between the thoracic and lumbar apices (relative apical distance) that will maintain acceptable balance (decompensation < or = 10 mm). Smaller values of the relative apical distance will decompensate the spine. For a constant postoperative thoracic Cobb angle, the postoperative distance between the thoracic and lumbar apices needed to maintain a balanced spine increases with increasing postoperative lumbar Cobb angle. Similarly, for a constant difference between the postoperative thoracic and lumbar Cobb angles, the postoperative distance between the thoracic and lumbar apices needed to maintain a balance spine increases with increasing postoperative thoracic Cobb angle. For postoperative thoracic curves of 20 degrees-30 degrees, acceptable balance can be achieved when the magnitude of the postoperative lumbar curve is up to twice the thoracic curve as long as adequate postoperative relative apical distance can be maintained. CONCLUSIONS: Decompensation does not appear to be caused by the relative magnitudes of the postoperative thoracic and lumbar curves, but is a result of inadequate relative distance between the thoracic and lumbar apical vertebrae in the postoperative geometry. PMID- 8727195 TI - Distractive properties of a threaded interbody fusion device. An in vivo model. AB - STUDY DESIGN: Twenty sheep underwent anterior lumbar interbody fusions with either a threaded titanium interbody fusion device (cage, n = 8), autogenous iliac crest dowel graft (autograft, n = 6), or interbody decortication only (sham, n = 6). Two sheep had misplaced cages and were excluded. Sheep were killed after 6 months. OBJECTIVES: To determine whether this model is useful for examining the distractive and fixation properties of interbody fusion cages. SUMMARY OF BACKGROUND DATA: Interbody fusion cages are used in anterior lumbar interbody fusion procedures to provide immediate intersegmental fixation and to distract and preserve interbody height. The process of physiologic anchorage by bone ingrowth into such devices is under investigation. METHODS: Sheep were radiographed immediately after surgery and 2, 4, and 6 months after surgery. Interbody distraction and angulation were measured with a digital photo image analyzer at each time point. After the sheep were killed, stiffness to flexion, extension, and lateral bending moments were measured. Twelve untreated cadaver spines were also tested for comparison. RESULTS: After surgery, interbody distraction successfully occurred in cage and autograft-implanted sites. Loss of interbody height ensued, however, in all groups during the first 2 months. Percentage loss of height was lowest in cage sites. By 6 months, only cage sites remained distracted beyond normal. Fusions in all groups were stiffer than untreated spines. Autograft sites were stiffer than cage sites to lateral bending. Sham sites were stiffer than cage and autograft sites to flexion, but this likely resulted from complete intervertebral collapse. CONCLUSION: Despite early subsidence, interbody fusion cages successfully distracted and preserved interbody spaces. This model is useful for investigating methods of improving distraction and stabilization. PMID- 8727196 TI - Biomechanical analysis of lumbosacral fixation. AB - STUDY DESIGN: The authors sought to measure the stiffness at the lumbosacral junction when it has been immobilized by means of two different posterior fixation systems in conjunction with three different anterior interbody fixation techniques. The information obtained provides a foundation for determining how methods of lumbosacral spinal fixation can maximize rigidity and improve fusion rates at this clinically important anatomic site. OBJECTIVES: To determine which spinal instrumentation systems, alone or in combination, provide the most stability at the lumbosacral junction. SUMMARY OF BACKGROUND DATA: In the clinical setting, several different posterior instrumentation systems are currently used, but the stability of these systems, when used in conjunction with anterior interbody fixation, has not been adequately tested. The authors hypothesized that rigidity of fixation at the lumbosacral junction may be enhanced by use of appropriate anterior interbody fixation techniques. METHODS: Two posterior and three anterior fusion constructs were tested in eight fresh frozen human cadaveric specimens in axial compression and torsion, flexion, extension, and lateral bending. RESULTS: The isolated posterior instrumentation constructs produced greater intervertebral stiffness than the isolated anterior constructs tested. The Jackson instrumentation was stiffer than the Galveston in axial torsion and flexion. Anteriorly, the femoral ring was stiffer than either the threaded interbody fusion device in flexion or the corticocancellous bone graft in flexion and compression. The threaded interbody fusion device was the stiffest anterior construct in axial torsion. CONCLUSIONS: The authors' data demonstrate that the threaded interbody fusion device may be an effective system for immobilization of the L5-S1 disc space. Rigidity of fixation at the lumbosacral junction may be enhanced by use of appropriate anterior interbody fusion techniques. PMID- 8727197 TI - Complications and long-term outcome of upper cervical spine arthrodesis in patients with Down syndrome. AB - STUDY DESIGN: A retrospective review of 15 patients with Down syndrome who had undergone arthrodesis of the upper cervical spine for instability. OBJECTIVES: To determine the complication rate and long-term outcome after posterior cervical arthrodesis for upper cervical instability in patients with Down syndrome. SUMMARY OF BACKGROUND DATA: Atlantoaxial instability is common in patients with Down syndrome, and fusion of the upper cervical spine has been recommended for patients who have instability, with or without myelopathy. Unfortunately, the results of posterior cervical arthrodesis are not well reported, and the natural history of this condition is unknown. METHODS: Fifteen patients with an average follow-up period of 74.6 months (range, 24-142 months) were reviewed after posterior arthrodesis of the upper cervical spine. Twelve patients were reexamined by the investigators specifically for the purpose of this study, and three patients had long-term follow-up results available from chart review. RESULTS: Eleven of 15 patients (73%) sustained 23 major complications including nonunion, loss of reduction, neurologic deterioration, late subaxial instability, infection, and wound dehiscence. Six patients (40%) required seven reoperations to address a complication. Ultimately, 12 patients (80%) obtained osseous union, but a definite clinical improvement was identifiable in only three patients, whereas two others had worsened neurologically at latest follow-up evaluation. CONCLUSIONS: A high complication rate should be anticipated after posterior arthrodesis of the upper cervical spine in patients with Down syndrome. A cautious approach to asymptomatic instability in this condition is advocated. PMID- 8727198 TI - Paraplegia resulting from vessel ligation. AB - STUDY DESIGN: This is a retrospective review of clinical records for evidence of paraplegia specifically resulting from segmental vessel ligation during anterior spinal surgery. OBJECTIVES: To determine the precise risk rate, and to potentially identify risk factors. SUMMARY OF BACKGROUND DATA: Although many authors have alluded to this risk, the exact risk rate and risk factors have never been identified. METHODS: All patients having an anterior approach involving T1-L3 were reviewed. The two reviewers were not involved in any of the surgeries. The 1197 cases were consecutive from 1967 to 1991. RESULTS: There were no paralyses. CONCLUSIONS: There would appear to be virtually no risk to segmental vessel ligation provided: 1) vessel ligation is unilateral, 2) done on the convexity of a scoliosis, 3) ligated at midvertebral body level, and 4) hypotensive anesthesia is avoided. Soft clamping with somatosensory-evoked potential monitoring does not appear justified. PMID- 8727199 TI - The congenital dislocated spine. AB - STUDY DESIGN: The congenital dislocated spine has been defined as the potentially most serious form of congenital kyphosis with an abrupt single-level displacement of the spinal canal. A retrospective chart review was conducted on 19 patients with this deformity. OBJECTIVES: To delineate the clinical and radiologic characteristics of this entity, and to analyze the outcome after treatment. SUMMARY OF BACKGROUND DATA: An anterior failure of formation was the basic feature. Kyphosis was variable. Vertebral displacement in the frontal plane was present in seven patients, and sagittal displacement was constant. Mechanical instability was seen in 17 patients. Neurologic impairment was identified in 12 patients, and congenital paraplegia was seen in eight patients. An acute paraplegia occurred after minor trauma in one patient. METHODS: Seventeen patients were treated surgically. Thirteen patients underwent complete circumferential stabilization through anterior strut grafting and posterior fusion without instrumentation, usually before age 3 years. Neurosurgical decompression was done in four patients. RESULTS: The average follow-up period was 8 years, 6 months. Nonunion of the posterior fusion mass was detected and successfully treated in five patients. A solid fusion seemed to be obtained in all patients at last follow-up evaluation. The neurologic status after neurosurgical decompression remained unchanged in three patients and was improved temporarily in one patient. CONCLUSIONS: Avoidance of neurologic morbidity requires early diagnosis and stabilization. The authors recommend early anterior strut grafting and posterior fusion. Exploration of the posterior fusion mass should be done systematically. PMID- 8727200 TI - The use of pedicle screw fixation to improve correction in the lumbar spine of patients with idiopathic scoliosis. Is it warranted? AB - STUDY DESIGN: A retrospective assessment of coronal, sagittal, and axial correction using convex lumbar pedicle screw constructs compared with hook constructs in patients with idiopathic scoliosis. OBJECTIVE: To determine if pedicle screw constructs can improve coronal, sagittal, and axial correction without increased complications and therefore warrant their use in the lumbar spine. SUMMARY OF BACKGROUND DATA: Although hooks have been the traditional fixation choice for posterior scoliosis correction of the lumbar spine, pedicle screws may offer advantages for improved correction of lumbar spinal deformity. METHODS: Twenty-two patients constituted Group A (hooks), in which 17 were double major and five were King Type IV curves. These patients had a minimum follow-up period of 2 years and an average of one hook per lumbar fusion segment. Twenty two patients constituted Group B (screws), in which 20 were double major and two were King Type IV curves. These patients had a minimum follow-up period of 2 years, and screw configuration consisted of pedicle fixation on the convex side for correction and at times on the concave side for fixation. RESULTS: Pedicle screw fixation constructs had improved lumbar Cobb correction (P < 0.05), lowest instrumented vertebra tilt (P < 0.05), lowest instrumented vertebra translation (P < 0.01), and segmental sagittal alignment from T12 to lowest instrumented vertebra (P < 0.01). There was no significant change in axial rotation using either surgical method. CONCLUSIONS: The use of pedicle screw fixation on the convex portion of the lumbar spine in patients with double major idiopathic scoliosis allows for improved correction of the lumbar Cobb measurement, horizontalization and translation of the lowest instrumented vertebra, and improved segmental lordization over the instrumented levels without increased complications. PMID- 8727201 TI - New anterior instrumentation for the management of thoracolumbar and lumbar scoliosis. Application of the Kaneda two-rod system. AB - STUDY DESIGN: The Kaneda multisegmental instrumentation is a new anterior two-rod system for the correction of thoracolumbar and lumbar spine deformities. This system consists of a vertebral plate and two vertebral screws for individual vertebral bodies and two semirigid rods to interconnect the vertebral screws. Clinical results of 25 thoracolumbar and lumbar scoliosis patients treated with this new instrumentation were analyzed. OBJECTIVES: To evaluate the efficacy of the new anterior instrumentation in correction and stabilization of thoracolumbar and lumbar scoliosis. SUMMARY OF BACKGROUND DATA: Since Dwyer first introduced the concept of anterior spinal instrumentation and fusion for scoliosis, anterior surgery has gradually gained acceptance. In 1976, a useful modification for the anterior spinal instrumentation, which reportedly provided means of lordosation and vertebral body derotation, was described. However, some authors reported a high tendency of the implant breakage, loss of correction, progression of the kyphosis, and pseudoarthrosis as the major complications. To overcome the disadvantages of Zielke instrumentation, the authors have developed a new anterior spinal instrumentation (two-rod system) for the management of thoracolumbar and lumbar scoliosis. METHODS: Anterior correction and fusion using Kaneda multisegmental instrumentation was performed in 25 patients with thoracolumbar or lumbar scoliosis. The average follow-up period was 3 years, 1 month (range, 2 years to 4 years, 7 months). There were 20 patients with idiopathic scoliosis (13 adolescents and seven adults) and five patients with other types of scoliosis, including congenital and other etiologies. All patients had correction of scoliosis by fusion within the major curve, and for 16 of the 25 patients, the most distal end vertebra was not included in the fusion (short fusion). Radiographic evaluations were performed to analyze frontal and sagittal alignments of the spine. RESULTS: The average correction rate of scoliosis was 83%. Over the instrumented levels, the correction rate was 90%. Preoperative kyphosis of the instrumented levels of 7 degrees was corrected to 9 degrees of lordosis. Sagittal lordosis of the lumbosacral area beneath the fused segments averaged 51 degrees before surgery and was reduced to 34 degrees after surgery. The trunk shift was improved from 25 mm before surgery to 4 mm at final follow-up evaluation. The average improvement in the lower end vertebra tilt-angle was 97% in those patients whose lower end vertebra was included in the fusion and 83% in patients whose lower end vertebra was not included in the fusion. Apical vertebral rotation showed an average correction rate of 86%. At final follow-up evaluation, all patients demonstrated solid fusion without implant-related complications. There was 1.5 degrees of frontal plane and 1.5 degrees of sagittal plane correction loss within the instrumented area at final follow-up evaluation. CONCLUSIONS: New anterior two-rod system showed excellent correction of the frontal curvature and sagittal alignment with extremely high correction capability of rotational deformities. Furthermore, correction of thoracolumbar kyphosis to physiologic lordosis was achieved. This system provides flexibility of the implant for smooth application to the deformed spine and overall rigidity to correct the deformity and maintain the fixation without a significant loss of correction or implant failure compared with conventional one-rod instrumentation systems in anterior scoliosis correction. PMID- 8727202 TI - The North American Spine Society (NASS) on lumbar discography. PMID- 8727203 TI - Performing anterior cervical fusion with either autograft or allograft. PMID- 8727204 TI - CGRP-LI. PMID- 8727205 TI - Distinctive features of the alpha 1-domain alpha helix of HLA-C heavy chains free of beta 2-microglobulin. AB - Only a few monoclonal antibodies are available with a restricted specificity to HLA-C products. In the present report, we demonstrate that antibody L31, previously shown to react with beta 2m-less (free) class I MHC heavy chains, binds to an epitope (residues 66-68 of the alpha 1 domain alpha helix) present on all the HLA-C alleles corresponding to the accepted (CW1 through CW8) serologic specificities, and on a few HLA-B heavy chains sharing with HLA-C an aromatic residue at position 67. Extensive IEF blot testing of HLA homozygous, EBV transformed B-lymphoid cells indicates that HLA-C molecules are present at significantly lower levels than HLA-B polypeptides not only at cell surface, as previously demonstrated, but also in total cellular extracts. Testing of metabolically labeled HLA-CW1, -CW5, and -CW6 transfectants and HLA homozygous lymphoid cells, particularly HLA-CW1-expressing cells, demonstrates that the L31 epitope is present on a subpopulation of naturally occurring HLA-C molecules distinct from that identified by antibody W6/32 to beta 2m-associated heavy chains. Pulse-chase experiments demonstrate that this epitope is transiently made available to antibody binding at early biosynthetic stages, but becomes hidden upon assembly with beta 2m. Thus, free HLA-C and other Y/F67+ heavy chains are characterized by distinctive antibody binding features in a region (residues 66 68) included in a previously identified HLA-C restricted motif, which has been suggested to be the primary cause of distinctive features of the antigen-binding groove, low affinity for endogenous peptide antigens and beta 2m, and preferential uptake of exogenous peptides, possibly of viral origin. We also show that HLA-CW1 heavy chains, both free and beta 2m associated, acquire sialilation. Free HLA-CW1 heavy chains are expressed at the cell surface even when unsialilated, albeit at low levels. PMID- 8727206 TI - CD48 may serve as an accessory molecule for the activation of a subset of human gamma/delta T cells. AB - To further assess the role of CD48 in the interaction of human gamma/delta T cells with their specific target, we generated two series of alloreactive clones, L and K. These clones express a V1-D-J1-C delta chain associated to V3-J2-C2 (L) or V2-J2-C2 (K) gamma chain. Functionally they were CTLs able to lyse the sensitizing B-cell line E418. The cytotoxicity of the L and K clones toward E418 was inhibited by anti-CD48 mAb. That of the L clones was also inhibited by anti HLA class I mAbs. Variation in L and K lysis profile was observed against a panel of CD48+ targets, further strengthening the argument that they display distinct specificities and suggesting that they do not recognize CD48. Heterogeneity in TCR gene segment usage, MHC-dependent recognition of E418 by the L clones, and resistance of some CD48+ targets strongly suggest that CD48 itself does not interact with L and K TCR. Transfection of CHO cells with CD48 induced killing by the K clones. This killing was inhibited by anti-CD48 mAbs. Taking into account the recent reports on CD48 as an accessory molecule, our results suggest that by binding to CD2 (and/or an unknown ligand), CD48 may serve to strengthen E/T interaction and may contribute to the activation of a minor subset of gamma/delta T cells. PMID- 8727207 TI - Identification of an epitope for T-cells correlated with antibody response to hepatitis B surface antigen in vaccinated humans. AB - Two antigenic T-cell epitopes of HBsAg, designated HBs 16-31 and HBs 81-99, were identified using synthetic peptides and HBsAg-specific T-cell lines. HBs 16-31 was recognized by five HBsAg-specific T-cell lines from vaccinees with both high and low antibody titers, whereas HBs 81-99 was recognized by two T-cell lines derived from vaccinees with high antibody titers. The antibody titer against HBsAg was correlated significantly with the proliferation of vaccinee's PBLs in response to HBs 81-99 (r = 0.47) but not to HBs 16-31, suggesting that HBs 81-99 plays a critical role in anti-HBs antibody production in humans vaccinated with HBsAg. PMID- 8727208 TI - Association of responsiveness to the major pollen allergen of Parietaria officinalis with HLA-DRB1* alleles: a multicenter study. AB - Parietaria, a plant belonging to the family of Urticaceae, is a major source of allergenic pollen in Europe. In the context of a multinational study, we investigated whether in allergic subjects antibody response towards Par o 1, the major allergen from P. officinalis, was associated with defined HLA-DRB1* alleles. The study population consisted of 234 allergic patients: 65 from Bulgaria, 30 from Israel, 99 from Italy, and 40 from Spain. In the Italian study group, the prevalence of ST positivity to Parietaria was 77%. In Parietaria ST positive subjects, the prevalences of IgG and IgE serum Ab towards Par o 1 were 91% and 75%, respectively. HLA-DRB1*1101 and/or 1104 were significantly positively associated with the presence of IgG Ab and with high levels of IgE Ab towards this allergen (p = 0.0007 and p = 0.012, respectively). In the Spanish study group, the positive association of DR1100 with responsiveness to Par o 1 was confirmed (p = 0.02, RR = 4, and p = 0.002, RR = 7, for IgG and IgE Ab, respectively). None of the Bulgarian patients had IgE Ab to Par o 1, whereas IgG Ab response was observed in 7 out of 65 subjects and was positively associated with DRB1*1101 and/or 1104 (p = 0.025). In the Israeli study group, responsiveness to Par o 1 was not associated with specific HLA-DRB1* alleles. In conclusion, this study shows that in allergic patients from three European populations antibody response to the major allergen from the pollen of Parietaria is associated with HLA-DRB1*1101 and/or 1104. Our data suggest that this association is stronger in subjects monosensitized to Parietaria. PMID- 8727209 TI - HLA-B40, B18, B27, and B37 allele discrimination using group-specific amplification and SSCP method. AB - We developed a system for discriminating HLA-B40, B18, B27, and B37 alleles using a two-step PCR method followed by SSCP analysis. Fragments (0.8 kb) including exon 2, intron 2, and exon 3 were amplified in the first PCR. We used two sets of primers, one specific for HLA-B60-related alleles and the other specific for HLA B61-related, B18, B27, and B37 alleles. No amplifications of other class I genes or pseudogenes were observed. In the second PCR, exon 2 and exon 3 were amplified separately, using diluents of the first PCR products as templates. HLA-B61 related, B18, B27, B37, and B60-related alleles were clearly discriminated in the SSCP analysis of the second PCR products. In a population study in which B61 alleles were analyzed, B*4003 was detected in two Japanese individuals in addition to two B61 alleles previously reported to occur in Japanese, B*4002 and B*4006. The relative frequencies of B*4002, B*4006, and B*4003 in Japanese were 58, 35, and 6%, respectively. The individuals having B*4003 are the first non South Americans in whom this allele has been detected. The SSCP banding patterns of 18 HLA-B60-positive Japanese population samples were identical to those of a B*40012 sample for both exon 2 and exon 3. We also demonstrated that the B37 allele occurring in some Japanese is B*3701. PMID- 8727210 TI - Genetic susceptibility and anti-human platelet antigen 5b alloimmunization role of HLA class II and TAP genes. AB - Platelet alloimmunization may result in post-transfusion purpura, and during pregnancy may cause neonatal alloimmune thrombocytopenia (NAIT), with a frequency estimated at 1.3 per 1000 live births. The risk of morbidity is significant: 20% of affected infants have neurologic sequelae and the death rate is about 10%. A better understanding of the immune response to platelet alloantigens would allow for a better definition, and thus better management of pregnant women at high risk. Limited data are available on the immune response against HPA-5b, the second most frequent antigen, after HPA-1a, implicated in NAIT. We studied HLA class II and TAP gene polymorphism in 50 women immunized against HPA-5 system antigens. Our results suggest a strong association of alloimmunization with a cluster of HLA DR molecules sharing a particular polymorphic amino acid sequence at position 69-70 (Glu-Asp encoded by GAA-GAC nucleotide sequence) of the DR beta 1 chain (RR = 2.95, RR = 5.70 when patients were homozygous for this sequence), and a negative association with the DRB1*0301 allele (2.1% vs. 28%; RR = 0.08). Furthermore, increased frequency of a TAP2 dimorphism at position 379 was observed in immunized women against the HPA-5 antigens (RR = 4.7). PMID- 8727211 TI - Simplified PCR-SSO procedure for DQA and DQB typing. Simultaneous hybridization of two probes to one membrane. AB - To simplify DQA and DQB oligotyping, we applied our improved PCR-SSO procedure for DR typing. We used 12 oligonucleotide probes for DQA typing and 18 for DQB typing. Oligonucleotide probes that require the same hybridization and stringent washing conditions were selected as pairs for simultaneous hybridization to a dot blot membrane containing various DNA samples. One probe of each pair was labeled with digoxigenin and the other with biotin. After hybridization, the dot-blot membranes were incubated with a mixture of conjugates. Specific binding of the corresponding DNA probes was visualized on an X-ray film using a chemiluminescent substrate (CSPD) and by staining using a chromogenic substrate (TMB). This approach, previously employed for DR typing, is also suitable for DQA and DQB oligotyping and significantly reduces the labor inherent in PCR-SSO typing. PMID- 8727212 TI - Monensin concentrations measured in feeder cattle using enzyme immunoassay. AB - Thirty heifers were fed a ration containing 30 g monensin/ton. Fecal, urinary and seral samples were collected at varying intervals prior to and after initiating administration of the monensin-containing feed, and monensin concentrations were determined using a modified indirect enzyme immunoassay. Fecal samples contained measurable (micrograms/g; ppm) concentrations of monensin in most samples. The majority of sera and urine samples contained monensin at ng/ml (ppb) concentrations, which were above background levels prior to monensin feeding. Twelve head were fed monensin at 60 g/ton and 90 g/ton for 5 d with collection of similar samples. Higher concentrations of monensin were detected with increasing ration amounts in all 3 sample types. Enzyme immunoassay for monensin in these biological samples identified presence of the feed additive. PMID- 8727213 TI - Metoclopramide and (or) cimetidine administration to sheep fed tall fescue. AB - Fifteen crossbred sheep were assigned randomly to 1 of 5 treatments: endophyte free fescue diet (EF; 0% infected; 50% hay, 40% seed, 10% molasses); endophyte infected diet (El; 95% infected; 50% hay, 40% seed, 10% molasses); El + 15 mg metoclopramide/kg body weight, 3 times/w; El + 9.0 mg cimetidine/kg body weight, twice daily; or El + metoclopramide + cimetidine. Both diets contained 92% dry matter (DM), 94% organic matter (OM) and 12% crude protein. All sheep were housed in individual metabolism stalls at 30 C, 14-h light:10-h dark for 18 d. Serum prolactin, respiration rate, heart rate or ear tip, coronary band and rectal temperatures were not affected by any of the treatments after 18 d (P > .10). No differences were detected in nutrient digestibilities except for treatment with metoclopramide + cimetidine, which produced decreased DM, OM and energy digestibilities. Sheep fed the El diet did not exhibit typical fescue toxicosis; therefore, the use of metoclopramide and(or) cimetidine to prevent toxicosis could not be evaluated. PMID- 8727214 TI - Genotoxicity studies of quercetin and shikimate in vivo in the bone marrow of mice and gastric mucosal cells of rats. AB - Bracken fern (Pteridium aquilinum) causes acute and chronic toxicity in animals and is associated with a high incidence of stomach and esophageal tumors in humans in countries where it is consumed as food, yet the toxic constituent(s) has not been unequivocally identified. None of the toxic constituents so far isolated from bracken have individually reproduced all the disease syndromes typical of the plant. The carcinogenicity of the plant has at one time or another been attributed to quercetin, shikimate or ptaquiloside. Studies to determine whether or not the genotoxicity of quercetin, shikimate (and its metabolite cyclohexane-carboxylate) could be enhanced in combined application were conducted in vivo using the micronucleus test in mice and the unscheduled DNA synthesis in rat gastric mucosal cells. None of these compounds were genotoxic; simultaneous administration of the compounds did not influence their in vivo genotoxicity. Plasma concentrations of quercetin after p.o. and i.p. administration at 200 mg/kg were below 0.1 microgram/ml, suggesting that the negative in vivo results may have been due to limited bioavailability of the compound. PMID- 8727215 TI - The effects of verapamil and lidocaine on the duration of propofol and xylazine ketamine anesthesia in broiler chickens. AB - The effects of verapamil, a calcium channel blocker, and lidocaine, a sodium channel blocker, administration on the duration of 1 mg propofol/kg i.v. or 5 mg xylazine/kg + 15 mg ketamine/kg i.m. anesthesia were examined in 1 and 6-w-old broiler chickens. The use of 2.5 or 1.25 mg verapamil/kg i.m. or 5 mg lidocaine/kg im prolonged the sleep time induced by the anesthetic agents. Administration of 5 mg verapamil/kg i.m. caused unexpected acute toxicity and mortality. This study suggests that propofol and xylazine + ketamine anesthesia in broilers may be modulated by calcium and sodium channels. PMID- 8727216 TI - Copper concentrations in sheep liver and kidney in Greece. AB - Copper liver and kidney concentrations were evaluated from sheep grazing at 2 areas: A (sampling points A1, A2, A3) and B. One hundred forty specimens were measured by flame atomic absorption spectroscopy after wet digestion for the February through April 1995 sampling period. Mean liver concentrations were: Area A (A1 = 99.48, A2 = 62.28, A3 = 23.02 mg/kg ww) and Area B 74.07 mg/kg ww. Copper concentrations in kidneys from all areas were in the normal range. PMID- 8727217 TI - The effect of lead on the bone marrow stem cells of mice infected with Listeria monocytogenes. AB - We investigated the effects of lead exposure on the growth and differentiation of bone marrow hematopoietic cells, the so called colony-forming cells, in normal and Listeria monocytogenes infected mice (resistant and susceptible strains). We also studied the effects of lead on the serum colony-stimulating activity (CSA), as well as on the survival of the mice after the infection. The doses of lead acetate were 13, 130 and 1300 ppm for 10, 30 and 70 d. At the end of this dosing, mice were infected with Listeria monocytogenes and killed 24, 48 or 72 h after inoculation of the bacteria. A dose-response suppressive effect of lead was observed in both strains in the 3 periods studied. However, in the resistant strain of mice the suppressive effects were overcome 48 h after the administration of the bacteria, whereas in the susceptible mice the suppressive effect of the infection was evident in all 3 time periods. The administration of lead caused no changes in serum hematopoietic growth factors, thus suggesting this metal acts by direct action on the myelopoietic cells. A significant decrease in host resistance, as measured by the mortality rate, was found when both strains of mice were challenged with sub-lethal doses of Listeria monocytogenes. Lethality was determined for a period of 10 d after dosing with 1300 ppm lead for 30 d. PMID- 8727218 TI - Lymphocyte esterases and hydroxylases in neurotoxicology. AB - Certain human lymphocytic enzymes, such as neuropathy target esterase (NTE), have become useful markers in clinical toxicology. NTE has been proposed as a predictive marker in organophosphate poisoning for the subsequent development of organophosphate-induced delayed neuropathy. We studied lymphocyte aryl hydrocarbon hydroxylases, and using a differential method based on that of NTE employing a phenyl-alkanoic substrate, developed an enzyme assay for use in toxic neuropathies. The assayed enzyme, which we refer to as neuropathy target hydroxylase (NTH), performed similarly to NTE in the evaluation of coherent clinical data obtained in patients with toxic neuropathies. This study indicates good correlation between the severity of clinical illness and abnormally low levels of NTH in neuropathies of varied etiology. A simplified protocol for NTH assay is presented. PMID- 8727220 TI - Experimental intoxication of sheep and cattle with Wedelia glauca. AB - Wedelia glauca was administered experimentally to 11 sheep and 4 cattle. The minimum toxic dose for both species was of 4 to 5 g of fresh plant/kg bw. Clinical signs were depression, muscle fasciculations, increased respiratory and cardiac frequencies, opisthotonous, sternal or lateral recumbency and terminal paddling movements. Time of onset of signs ranged from 12 to 40 h after dosing. Serum AST, LDH and GGT were increased. Three cattle and 5 sheep died after clinical manifestation periods of 2 to 18 h, and 2 sheep survived after being affected for 14 and 46 h. Macroscopic and histologic lesions were similar in cattle and sheep; the liver was swollen and dark reddish, and the wall of the gall bladder was edematous. The cavities had yellowish fluid. Petechiae and echymoses were seen on serous membranes. Microscopically the liver had periacinar hemorrhagic necrosis. Two sheep dosed with 1 g/kg bw daily for 20 d and 1 dosed with 1 g/kg bw for 10 d were not affected. Six sheep were transferred from an area free of W glauca to an area where this plant was present. These animals ate small amounts of the plant and lost weight rapidly, but clinical intoxication did not occur. Two sheep were fed with lucerne hay containing 9% W glauca. They ingested 2.5 g/kg of the dry plant (corresponding to 10 g/kg bw of fresh plant) in 24 h, but did not show intoxication. Because the intoxication occurred in sheep and cattle administered 4 to 10 g/kg bw of the plant in a period of 1 to 2 h, the intoxication may only occur when animals ingest a single toxic dose in a short time. PMID- 8727219 TI - Effects of selected organophosphate insecticides on serum cholinesterase isoenzyme patterns in the rat. AB - The serum cholinesterase (ChE) isoenzyme in rats shows 6 bands after polyacrylamide gradient gel electrophoresis. The effects of organophosphates (fenthion, chlorpyrifos, diazinon, bromophos, propaphos, haloxon, and DFP) on serum ChE isoenzyme bands were studied in 32 male and 32 female 6-w-old Sprague Dawley rats. Each organophosphate was randomly administered to 4 male and 4 female rats. Blood samples were collected from the abdominal aorta under halothane anesthesia 6 h after dosing. The isoenzyme patterns were determined simultaneously with erythrocyte and serum ChE activities. Changes were observed in all 6 bands of the serum ChE isoenzymes after administration of fenthion, chlorpyrifos and propaphos. Diazinon had no influence on band 6, and DFP and bromophos had no influence on band 5. Haloxon did not effect any of the serum ChE isoenzyme bands. Serum ChE was most suppressed by fenthion, followed by DFP, bromophos, chlorpyrifos, propaphos and diazinon in that order of effect. Serum ChE activity was not suppressed by haloxon. Erythrocyte ChE activity was suppressed by every organophosphate. This experiment demonstrated a correlation between the organophosphate suppression of serum ChE activity and the concentration of serum ChE isoenzyme band 6. PMID- 8727221 TI - Heavy metal concentrations in injured racehorses. AB - Records were reviewed of horses that had fatal injuries at California racetracks over a 16-mo period. Horses were categorized based on injury type, sex, age and breed. Arsenic, cadmium, copper, iron, lead, molybdenum and zinc concentrations were measured in the liver and kidneys from each horse. Arsenic and lead were not detected in any tissues. Liver heavy metal concentrations were not related to the injury type. Kidney iron concentrations were significantly (P < 0.05) higher in horses with ligament ruptures and in horses with fractured sesamoid, carpus or metacarpus/metatarsus bones. Liver cadmium, kidney cadmium, and liver iron were higher (P < 0.05) in horses > 2 y old than in 2-y-olds. Liver cadmium concentrations were higher (P < 0.05) in Thoroughbreds than in Quarter Horses. PMID- 8727222 TI - Subchronic study of the metabolism of triphenyltin in hamsters. AB - In previous studies we investigated the metabolism of tin compounds in rats and hamsters following single oral treatments with triphenyltin. The objective of this study was to provide information on the metabolism of triphenyltin in hamsters subchronically treated with dietary triphenyltin for 180 d. Groups were fed diets containing 3 different triphenyltin concentrations: 1.28, 28.82 or 54.77 ppm. The 28.82 and 54.77 ppm groups gained less body weight than the control and 1.28 ppm groups. However, none of the animals showed characteristic symptoms. We detected triphenyltin and its metabolites in the tissues of dosed hamsters. Highest tin concentrations were in livers and kidneys, as diphenyltin and inorganic tin respectively; no significant amounts of triphenyltin were found. Triphenyltin was metabolized relatively rapidly in the hamsters. There was more tin relative to the administered triphenyltin in the livers of the low dietary group compared to the high dietary triphenyltin group. This suggests that low concentrations of triphenyltin are easily absorbed from the gastrointestinal tract. While we previously demonstrated that acutely dosed triphenyltin produced marked hyperglycemia in hamsters, this was not found in the hamsters repeatedly dosed with dietary triphenyltin in the present study. PMID- 8727223 TI - A report of suicide involving verapamil. AB - A case of fatal poisoning of an adult man with verapamil, the active component of ISOPTIN, is presented. Quantitative analyses were carried out using HPLC and GC, and results were compared. Verapamil was determined in stomach and liver specimens and in blood. A remarkably high concentration of this drug was found in the blood sample. PMID- 8727224 TI - Pesticide poisoning of animals of wild fauna. AB - Poisoning of rare birds of prey (7 Aegipius monachus and 1 Aquila chrysaetus) and 11 foxes by carbofuran is reported. The poisoning is an ecological disaster because of the death of A monachus, which is a rare species. Identification, confirmation and distribution of the toxic substance was performed by TLC and HPLC techniques. PMID- 8727225 TI - State of the nation's poison centers: 1994 American Association of Poison Control Centers survey of US poison centers. PMID- 8727226 TI - Pulmonary dysfunction in survivors of acute paraquat poisoning. AB - Five patients survived acute paraquat ingestion, despite developing restrictive pulmonary dysfunction. Of these, 2 patients with documented long-term follow-up had progressive functional improvement. A literature review revealed 29 other patients surviving restrictive pulmonary dysfunction following acute paraquat poisoning. Some patients who survive an acute paraquat poisoning may develop pulmonary fibrosis, yet progressively improve over time. PMID- 8727227 TI - Outpatient N-acetylcysteine treatment for acetaminophen poisoning: an ethical dilemma or a new financial mandate? AB - The mainstay of treatment for acetaminophen-induced hepatotoxicity, produced by the accumulation of the toxic metabolite N-acetylbenzoquinoneimine, is an enteral 18-dose course of N-acetylcysteine (NAC). However, absence of characteristic symptomatology is a frequent reason for premature cessation of NAC and early discharge of the toxic acetaminophen poisoned patient. We report a series of confirmed acetaminophen poisonings who were discharged early with NAC and instructions to self-administer. All cases of acute acetaminophen poisoning without concomitant drugs, reported to a certified Regional Poison Information Center for a 3-mo period of time, were reviewed. Inclusion criteria included patients who were discharged with orders to complete the course of NAC outside of a hospital, despite toxic serum acetaminophen concentrations. Data parameters evaluated included age, amount taken, symptoms, laboratory results, treatment, and medical outcome. 131 cases of confirmed toxic acetaminophen poisoning yielded 6 patients who received 4 to 6 doses of NAC during hospitalization, but were discharged to home with the remaining 11-13 doses. Patients' ages ranged from 16 28 y (mean 20.0 y). Serum acetaminophen concentrations measured at 4 h post ingestion ranged from 171-198 mcg/ml (mean 182 mcg/ml). Follow-up by the certified Regional Poison Information Center at 1-3 w post-discharge determined dosing compliance to be 83%. All 6 patients remained asymptomatic with normal liver function testing. Since health care reform encourages practitioners to reconsider established approaches to the delivery of health care, perhaps home delivery of NAC would not only be clinically preferred to premature cessation of the antidote, but also offer cost savings. Self-administration of NAC in the home setting may be representative of a new era in America's health care delivery system. PMID- 8727228 TI - Meadow saffron (Colchicum autumnale) intoxication in a nomadic Albanian sheep flock. PMID- 8727229 TI - Polydrug use in an inpatient treatment sample of problem drinkers. AB - Over the past 30 years in the United States, there have been marked secular increases in polydrug use. Alcohol and other substance use disorders are highly comorbid. Yet, little research has characterized patterns of polydrug use in persons with alcohol dependence. In particular, little is known about this population's use of alcohol and other drugs in combination or on the same day, which is termed simultaneous polydrug use (SPU). This research assessed patterns of SPU in 212 problem drinkers who participated in an alcohol treatment outcome study. Subjects were given a Time-Line Follow-Back interview that assessed the use of alcohol and nine other drug classes for each day of the 120 days before treatment entry. A majority of subjects (61%) reported SPU during this assessment interval. Subjects who reported SPU were disproportionately younger, male, and unmarried, compared with those who did not report SPU. The most common alcohol/drug combinations were alcohol with cocaine (60% of subjects who reported SPU), alcohol with marijuana (51% of SPU subjects), and alcohol with sedatives (31% of SPU subjects). The most common three-drug combination was alcohol, cocaine, and marijuana (23% of SPU subjects). Alcohol use and drug use were associated at the event level, significantly more than association predicted by the base rates of the individual behaviors. Time-Line Follow-Back data correlated highly with a questionnaire measure of SPU. Results indicate that polydrug use is an important focus for assessment and intervention in alcohol treatment programs. PMID- 8727230 TI - Failure to find exon 7 polymorphism of the ADH7 gene in Chinese, Japanese, African-Americans, and Caucasians. AB - Class IV alcohol dehydrogenase (sigma-ADH) activity has been found in high levels in the stomach and esophagus, but not in liver. Gastric ADH activity has been reported to influence blood alcohol levels after oral ethanol ingestion, suggesting that sigma-ADH activity plays a role in first-pass metabolism. It has also been reported that women have lower sigma-ADH activity than men and that Asians have lower sigma-ADH activity than Caucasians and African-Americans. A genetic basis for these gender and ethnic differences in sigma-ADH activity has been postulated. A recent study in a Japanese subject found a point mutation in the codon for amino acid 287 of the ADH7 gene (which encodes sigma-ADH), changing the amino acid from glycine to valine. A polymerase chain reaction-sequencing assay was established to determine the frequency of this polymorphism in the Asian, Caucasian, and African-American populations. The polymorphism was not present in the 21 Asians, 15 Caucasians, and 3 African-Americans we genotyped, suggesting that if this polymorphism exists, its frequency is low in these ethnic groups. It is therefore unlikely to be responsible for the absence of sigma-ADH activity in gastric specimens from Asians. PMID- 8727232 TI - High alcohol-related premature mortality in France: concordant estimates from a prospective cohort study and national mortality statistics. AB - This study examines the magnitude of alcohol-related premature death in the French population, which still has the highest average alcohol intake in the world and a relatively low coronary heart disease mortality rate. Two data sources were used: the national mortality data in 1990 and a prospective mortality experience in a cohort of 2,687 middle-aged working men examined in 1980-1985 and followed-up during an average of 9.3 years. In the general population study, alcohol-related premature mortality (35-64 years) was calculated using alcohol-attributable fractions (AAFs) derived from studies of alcohol involvement in deaths from various causes. In the cohort, it was estimated from AAFs and attributable risk using both alcohol exposure prevalence and relative risks of death according to alcohol intake categories. In 1990, estimates of 19.1% and 13.0% of all premature deaths in French men and women were attributed to alcohol. Digestive diseases followed by malignant neoplasms in men and by unintentional injuries in women were major contributors to the total number of alcohol-related premature deaths. In the cohort study, 90 deaths occurred during the follow-up period. The estimation of alcohol-related premature mortality using AAFs was 24.4%. The relative risk of total mortality (adjusted for age, smoking habits, and body mass index) for men who consumed > 60 ml/day of alcohol, compared with those who consumed 0-25 ml/day was 1.9 (95% confidence interval: 1.1-3.5). The consumption of 26-60 ml/day was not associated with reduced risk of mortality, and the adjusted relative risk for this group was 1.3 (95% confidence interval: 0.7-2.5). The estimated attributable risk of premature mortality caused by consumption of > 25 ml/day of alcohol was 29.9%. This study reports the persisting high alcohol-related premature mortality in the French general population, as well as in middle-aged working men. The results suggest that efforts should be paid to reduce further the consumption of alcohol in France. PMID- 8727231 TI - Relationships between depressive symptoms, anxiety, alcohol consumption, and blood pressure: results from the CARDIA Study. Coronary Artery Risk Development in Young Adults Study. AB - OBJECTIVES: We examined cross-sectional relationships among depressive symptoms, anxiety, alcohol intake, and blood pressure. Test hypotheses were that: (1) alcohol intake, depressive symptoms, and anxiety would be positively related to blood pressure; (2) depressive symptoms and anxiety would have a stronger association with alcohol intake in Blacks than in Whites; and (3) adjustment for differences in depressive symptoms, anxiety, and alcohol intake would reduce Black-White blood pressure differences. METHODS: Study hypotheses were tested in a sample of 4,352 Black and White adults, participating in the CARDIA study. Hypotheses were tested using multiple linear regression. RESULTS: Alcohol intake was positively related to systolic (p = 0.0001) and diastolic (p = 0.0004) blood pressure in men, but not in women. Depressive symptoms and anxiety were unrelated to blood pressure. The relationship between alcohol intake and depressive symptoms differed by race/ethnicity in men (p = 0.0719) and in women (p = 0.0002). Alcohol intake increased with increasing levels of depressive symptoms, but the increase was greater in Blacks than in Whites. After accounting for alcohol intake, body mass index, and other variables, Black-White blood pressure differences were reduced in men, but not in women; most of the reduction was caused by body mass index. CONCLUSIONS: Blacks may respond differently than Whites to psychological distress. PMID- 8727233 TI - Assessing alcohol problems in female DWI offenders. AB - The severity of alcohol problems in a sample of 812 women driving while intoxicated (DWI) offenders enrolled in a Drinking Driver Program in New York State was assessed through the use of multiple measures. These women voluntarily provided information through a self-administered questionnaire completed at the beginning of the program. Classification of alcohol problems based on DSM-III-R criteria was compared with classification based on the use of alcohol-specific and nonalcohol-specific measures of alcohol-related problems. Greater hostility and a larger number of drinks ever consumed accounted for nearly 12% of the variance between no diagnosis and alcohol abuse women, whereas these two variables plus age, race, depression, and having a relative with alcohol problems accounted for nearly 41% of the variance between alcohol abuse and dependent women. A subsample of women within each DSM-III-R diagnostic group showed high risk for later, more severe alcohol problems. Multiple measures of alcohol problems help to elucidate subtle differences in alcohol problems within each of the broad categories specified by the DSM, and should facilitate more appropriate intervention and treatment plans for women DWI offenders with alcohol-related problems. PMID- 8727234 TI - Has awareness of the alcohol warning label reached its upper limit? AB - Has awareness of the alcoholic beverage warning label reached its maximum? This study tracks changes in the level of awareness among a sample of 7334 inner-city African-American gravidas seeking prenatal care between May 1989 and June 1993. Previously, we found that a significant increase in awareness of the warning label occurred in March 1990. In the current analysis over a 50-month period, the level of awareness continued to increase through December 1992 and then leveled off, suggesting a negatively accelerated growth function. The logistic function fitted to the awareness curve predicts that the upper limit of awareness in this population has been reached (the predicted upper limit being 81.5%). In addition a logit regression analysis showed that women who did not know about the warning label were more likely to be over 29 years of age. Heavier drinkers were 1.25 times more likely to be aware of the label. Among those drinkers who were not aware of the label, 30% drank at both conception and antenally, thus putting their fetus at high risk for alcohol-related birth defects. PMID- 8727235 TI - Tridimensional personality traits in sons of alcoholic and nonalcoholic fathers. AB - Sons of alcoholics (SOAs; n = 27) and sons of nonalcoholics (SONAs; n = 23) were compared across Tridimensional Personality Questionnaire (TPQ) and Sensation Seeking scales and measures of plasma homovanillic acid and platelet monoamine oxidase activity. SOAs and SONAs did not differ significantly on any measure. The pattern of correlations between TPQ and Sensation Seeking scales provided some support for the construct validity of TPQ measures. Scores on the TPQ Novelty Seeking scale and platelet monoamine oxidase activity were significantly inversely associated (r = -0.52, p < 0.02) among SOAs, but not among SONAs (r = 0.06). PMID- 8727236 TI - Sensitivity and specificity of carbohydrate-deficient transferrin as a marker of alcohol abuse are significantly influenced by alterations in serum transferrin: comparison of two methods. AB - Despite a number of investigations suggesting the value of carbohydrate-deficient transferrin (CDT) as a marker of alcohol abuse, a variety of issues on the applicability of CDT measurements in clinical settings have remained unexplored. Earlier studies in this field have focused on the relationship of CDT and the amount of alcohol consumption or presence of liver disease, whereas the influence of alterations in serum transferrin concentrations on CDT has received less attention. In this study, we compared two different methods for measuring CDT (CDTect and %CDT) and total transferrin concentrations in a sample of 83 alcohol abusers (20 patients with alcoholic liver disease and 63 heavy drinkers who were devoid of liver disease, despite excessive alcohol consumption) and 89 controls, who were social drinkers or abstainers. The control population included 53 hospitalized patients with expected abnormalities in serum transferrin concentrations caused by conditions such as negative iron balance, pregnancy, or nonalcoholic liver disease. Both methods gave significantly higher values in alcohol abusers than in controls (p < 0.01), but the overall sensitivity for detecting alcohol abuse was clearly higher for CDTect (59%) than for %CDT (34%). The correlation between the results obtained by the two methods (r = 0.629) significantly improved, when the CDTect values were replaced by the ratio of CDTect/total transferrin (r = 0.770) (p < 0.05). There was a positive correlation between the CDTect and serum transferrin (r = 0.201, p < 0.01), which was significant both in the alcoholics (r = 0.240, p < 0.05), and especially in the controls (r = 0.727, p < 0.001). A significant inverse correlation emerged between %CDT and total transferrin (r = -0.302, p < 0.01). The sensitivities of CDTect and %CDT for correctly classifying alcohol abusers in the subgroup of alcoholic liver disease patients were 90% and 70% and in the subgroup of heavy drinkers without liver disease (49% and 22%), respectively. Specificities for CDTect and %CDT in this sample were 81% and 100%, respectively. However, in the subgroup of hospitalized control patients with abnormal serum transferrin, the specificity of CDTect was only 48%. According to present data, CDTect seems to be more sensitive than %CDT for detecting alcohol abuse. However, any alteration in serum total transferrin concentration markedly decreases the assay specificity. This should be considered when interpreting the assay results in patients with elevated serum transferrin, such as iron deficiency, pregnancy, or liver diseases. PMID- 8727237 TI - Prenatal exposure to alcohol and marijuana: effects on motor development of preschool children. AB - Gross motor development of preschool children prenatally exposed to alcohol and marijuana was assessed as part of a longitudinal study. Most mothers in the study were light to moderate users and discontinued or decreased use of alcohol and marijuana after the first trimester of pregnancy. The women were of lower socioeconomic status, half of the sample was African-American, and most were single. Gross motor development was evaluated with balance and ball-handling items at 3 years. Balance items included walking on a line, walking on a balance beam, standing on one foot, standing on tiptoes, and stair climbing and descent. Ball-handling items included catching, throwing, and kicking a ball. Refusal to perform items was also recorded. Prenatal alcohol and marijuana exposure did not negatively affect gross motor development. The composite score on the Stanford Binet Intelligence Scale, age at assessment, gender, and examiner were significant predictors of gross motor performance and of refusal to participate in the balance items. The ponderal index, number of siblings, current income, examiner, current maternal use of tranquilizers, and first trimester exposure to amphetamines were also significant predictors of balance skills. Gender and number of hospitalizations predicted refusal to participate in balance items, whereas hearing and vision problems predicted refusal on ball-handling items. The components of timing, speed, and fine motor control have not been addressed in this study, and therefore it is premature to conclude that there is no impact of prenatal substance use on motor development. PMID- 8727238 TI - Changes in human plasma nerve growth factor level after chronic alcohol consumption and withdrawal. AB - Numerous studies reported in recent years have shown that withdrawal from chronic consumption of drugs induces high levels of anxiety, both in humans and in animal models. In the present study, we demonstrated that withdrawal from chronic consumption of either ethanol or heroin causes a significant increase in plasma nerve growth factor, suggesting that the resulting anxiety condition triggers the release of this molecule. Although the functional significance of this phenomenon needs to be better defined, it is hypothesized that the increased levels of circulating nerve growth factor might be involved in homeostatic adaptive and/or reparative mechanisms. PMID- 8727239 TI - Effects of gender and comorbidity on problem drinking in a community sample. AB - We investigated the influence of gender, comorbidity, drinking history, and age on the clinical manifestations of DSM-III alcohol abuse and/or dependence in men and women. The sample was drawn from the National Institute of Mental Health Epidemiologic Catchment Area study, a large-scale, multicenter survey to investigate psychiatric disorders in the community. The results showed that gender and comorbidity had independent effects on problem drinking after drinking history and age had been taken into account. Gender contributed to the age of onset of problem drinking and the rate of its development. Comorbidity, drinking history, and age contributed independently to its severity. The effects of these variables in this community sample paralleled those reported in treatment samples. PMID- 8727240 TI - Pharmacological treatment for antisocial personality disorder alcoholics: a preliminary study. AB - A short time ago, we reported that a subgroup of hospitalized alcoholic men with comorbid antisocial personality disorder (ASP) seemed to benefit significantly from antidepressant medication at the end of a 6-month period in a double-blind, random assignment, placebo-controlled study. In a reanalysis of those data, we divided the ASP alcoholic group (n = 29) into those who did (n = 15) and who did not (n = 14) also satisfy DSM-III-R criteria for an additional current mood and/or anxiety disorder and then compared the 6-month outcomes of these two smaller subgroups. Despite the small ns, the results for most drinking outcome measures indicated: (1) that ASP alcoholics with a current mood/anxiety disorder improved significantly more with pharmacological treatment, relative to placebo; and (2) that ASP alcoholics with no current mood/anxiety disorder failed to respond differentially to pharmacological treatment over the 6-month period. These findings suggest a possibly useful and inexpensive approach to the long term management of a very difficult-to-treat subgroup of men substance abusers. PMID- 8727241 TI - A prospective, high-risk study of the relationship between tobacco dependence and alcohol use disorders. AB - This study examined the extent to which tobacco dependence (TD) and alcohol use disorders (AUDs) reciprocally influenced each other in a mixed-gender sample of 452 individuals (n = 232 biological family history of paternal alcoholism, n = 220 no first- or second-degree family history of alcoholism) who were assessed once early in their freshman year of college, approximately 3 years later when many were college seniors, and approximately 3 years later when many had entered or were entering the work force. AUDs were more prevalent in men than women, in individuals with a family history of alcoholism, and decreased overall with time. TD was more prevalent in those with a family history of alcoholism, showed increasing rates of use over time, and was less prevalent but more stable than AUDs. Transitional probabilities indicated that although a previous AUD or TD diagnosis increased the likelihood of being diagnosed with the other disorder at a later time, comorbid AUDs and TD did not significantly affect the likelihood of recovery from either disorder. Finally, path analysis revealed significant reciprocal relationships between AUDs and TD diagnoses (each predicting the other over time), and significant prediction of AUDs and TD by family history of alcoholism at the first and third times of assessment. Findings supported two general models of AUD/TD comorbidity: a shared vulnerability model and a reciprocal influence model. PMID- 8727242 TI - Alcohol and aldehyde dehydrogenase genotypes and drinking behavior in Japanese. AB - The effects of the genotype of alcohol dehydrogenase-2 (ADH2) and mitochondrial aldehyde dehydrogenase (ALDH2) on drinking behavior were investigated in a population of 451 Japanese. Although the ALDH2*2 allele had a significant inhibitory effect on alcohol consumption, hence on drinking problems, the apparent association was not confirmed between ADH2 genotype and overall drinking patterns for either males or females. However, the frequency of the ADH2*2 allele was significantly lower in male Japanese classified as alcoholic on the basis of the Kurihama Alcoholism Screening Test than in nonalcoholic males. These results corroborate a previous study that revealed a significantly lower ADH2*2 allele frequency in hospitalized Japanese alcoholics than in the general population. Together, these studies suggest that the ALDH2*2 allele has an inhibitory effect on drinking behavior, irrespective of the level of alcohol consumption, whereas the effect of the ADH2 polymorphism only becomes apparent in individuals with higher alcohol consumption, such as alcoholics. PMID- 8727244 TI - Voice of the victims. AB - Over the past 10 years, I have been privileged to conduct educational forums for audiences containing many recovering alcoholics or otherwise chemically dependent persons. In these forums about the addictive diseases and their treatment and research possibilities, significant interaction with the audience members occurs. During these interactions, certain anecdotal phenomena seem to predominate. The repetitive nature of these reports suggests the need for systematic investigation. As with editorial comments in major medical journals, observed phenomena and unanswered questions from the victims can be valuable in the generation of testable hypotheses. Perhaps the ideas presented herein will be useful in the development of future research on alcohol abuse and alcohol dependence. PMID- 8727243 TI - Tetrahydroisoquinolines and alcoholism: where are we today? AB - The present status of research on the tetrahydroisoquinoline (THIQ) family of compounds and other aldehyde metabolites in the field of alcoholism is described. A brief history of the background of experimental studies on the actions of the THIQ's and beta-carbolines on alcohol drinking is presented. A computer data base search of articles published in this field reveals that both the historical and current research trends have waxed and waned since the 1960s. The clinical utility of naltrexone in terms of the cerebral function of opioid compounds in drinking behavior and alcoholism is likewise considered. Finally, the residual controversy concerning the significance of multiple intermediary metabolites in alcohol dependence and craving should ultimately be resolved in the future by broad-based investigations which employ state-of-the-art experimental approaches. PMID- 8727245 TI - Postbinge effects of acute alcohol intoxication on hepatic free radical formation. AB - The present studies were performed to test the hypothesis that Kupffer and endothelial cells are activated after recovery from an acute alcohol binge, which is accompanied by formation of oxygen-derived radicals. These radicals have been implicated in the pathogenesis of alcohol-mediated tissue injury in a number of organs. Male Sprague-Dawley rats received an intravenous injection of 20% ethanol in saline (1.75 g/kg), followed by an intravenous infusion (250 to 300 mg/kg/hr) for 12 hr. At the end of 12-hr infusion, ethanol was replaced by saline, and the infusion was continued for a further 6 hr. This was referred to as the recovery period. The 6-hr recovery period was selected because superoxide anion generation by the perfused liver peaked at this time point. Superoxide anion formation by the perfused liver was measured by the superoxide dismutase-inhibitable reduction of ferricytochrome c. Kupffer and endothelial cells were isolated for the determination of in vivo glucose uptake and in vitro superoxide anion release. Results show that a significant (p < 0.05) amount of superoxide (1.54 nmol/min/g) was generated by the perfused liver at 6 hr recovery after 12 hr of ethanol infusion. Serum ALT activity was also elevated in this treatment group. Time matched control-saline infused animals or ethanol-treated animals without a recovery period released < 0.2 nmol/min/g of superoxide. The postrecovery superoxide production and an accompanying increase in the in vivo glucose uptake were also observed in isolated Kupffer and endothelial cells. Depletion of Kupffer cells by gadolinium chloride before ethanol treatment and recovery was associated with significant attenuation of free radical formation by the perfused liver and reduction of serum ALT. These studies demonstrate that recovery from an acute alcohol binge has a stimulating effect on hepatic sinusoidal superoxide production, and it may also affect liver function. PMID- 8727246 TI - Inhibitory mechanism of intestinal ethanol absorption induced by high acetaldehyde concentrations: effect of intestinal blood flow and substance specificity. AB - This study describes the effects of high blood acetaldehyde concentrations on the intestinal absorption of ethanol and 2-butanone using an in situ single-pass perfusion technique on the rat intestine and the colored microsphere method to measure intestinal blood flow. We found that high blood acetaldehyde concentrations inhibit intestinal ethanol absorption in an inverse proportion to peak acetaldehyde concentrations, decrease intestinal blood flow, and inhibit intestinal absorption of 2-butanone. The decrease of the intestinal blood flow, induced by high blood acetaldehyde concentrations, is a major mechanism inhibiting intestinal ethanol absorption, but other mechanisms are also thought to inhibit absorption. Therefore, inhibition by high acetaldehyde concentrations is not the only factor affecting ethanol absorption. PMID- 8727247 TI - Effects of maternal ethanol consumption on the subsequent development of immunity to Trichinella spiralis in rat neonates. AB - The immune response of rat pups to the intestinal parasite Trichinella spiralis was studied to determine if maternal pre- and/or postnatal ethanol consumption affected neonatal immune responses. Female rats were fed ethanol-containing (36% of calories) or pair-fed control liquid diets and include groups that were maintained on ethanol as follows: group 1, from day 1 of pregnancy through weaning and whose pups were then placed on ethanol to sacrifice; group 2, from day 1 of pregnancy through lactation; group 3, from day 1 of pregnancy through pup delivery; and group 4, from day 1 of lactation through weaning. A parallel group of animals was pair-fed isocaloric control diet until sacrifice. The pups of all litters were immunized orally with 500 L1 (T. spiralis) larva 5 days after weaning. To examine the effects of maternal ethanol on primary immune responses, one-fourth of the pups from each litter were sacrificed on days 10 and 20 after immunization. To examine the effects on neonatal secondary immune responses, the remaining pups were challenged with 1,000 larva 30 days after the initial immunization and sacrificed either 3 or 8 days after challenge. At the time of sacrifice, blood samples were collected, the intestine removed to determine T. spiralis worm burdens, and suspensions of mesenteric lymph node (MLN) cells prepared. Intestinal worm counts and serum levels of anti-T. spiralis IgM and IgG antibodies, interleukin-2 (IL-2), and tumor necrosis factor (TNF) were determined. In vitro proliferation responses of MLN cells to T. spiralis antigen and to the mitogen concanavalin A (Con A) were also examined. Pups from groups 1 to 3 demonstrated significantly higher intestinal worm counts (decreased immunity) than the pair-fed controls at the day 20 primary immune response sacrifice, and pups from group 1 had significantly higher worm counts at day 3 after a secondary immune challenge. Pups of dams from groups 1, 3, and 4 had significantly lower IgM antibody titers at the day 20 primary immune response sacrifice. All experimental ethanol groups (1 to 4) demonstrated significantly lower IgG antibody titers than that observed in pair-fed control pups at the 20 day sacrifice. IgM antibody titers showed significant reductions for ethanol treated groups at 3 and 8 days after T. spiralis secondary challenge. In addition, IgG antibody titers were also significantly reduced for all alcohol groups at 3 and 8 days during the secondary immune response. Serum IL-2 and TNF levels were significantly lower in all experimental ethanol groups (1 to 4) relative to pair-fed controls at day 20 during a primary immune response, and IL 2 levels at 3 days postchallenge were lower in groups 2 to 4 after a secondary immune challenge. MLN proliferation responses to antigen and Con A were significantly reduced in ethanol groups 1 to 3 and to Con A in group 4 at day 10 after a primary immune challenge. Ethanol group 3 pups also demonstrated a reduced response to antigen at day 20. For animals undergoing a secondary immune response, ethanol group 2 demonstrated a reduced response to antigen at day 3, whereas groups 2 and 4 showed increased reactivity to antigen at days 3 and 8 postchallenge. These results show that maternal ethanol consumption diminishes the capacity of neonates to respond to T. spiralis antigen and that the depressed immune response involves T- and B-cell-mediated reactions and also affects the production of certain cytokines. These results also suggest that the diminished immune responses are increased with longer periods of maternal and neonatal exposure to ethanol. PMID- 8727248 TI - Defective calcium increase and inositol phosphate production in anti-CD3 stimulated lymphocytes of alcoholics without progressive liver disease. AB - Intracellular free calcium concentration, phosphoinositide turnover, and inositol phosphate production were analyzed in peripheral blood lymphocytes from seven well-nourished alcoholic patients without severe acute or chronic liver disease, before and after stimulation with anti-CD3 antibody. Seven comparable nondrinkers were studied as controls. A lower increase in intracellular free calcium concentration was detected in alcoholics, after anti-CD3 stimulation of lymphocytes, than in control subjects. Lymphocyte activation generated inositol phosphates in both controls and alcoholics, but inositol phosphate production was significantly lower in alcoholics. The agreement between these findings indicates that the reduction in inositol phosphates is one of the most important events in the early phases of lymphocyte activation in alcoholics. PMID- 8727249 TI - No association of the structural dopamine D2 receptor (DRD2) variant 311Cys with alcoholism. AB - The human dopamine D2 receptor (DRD2) has been implied in the vulnerability for alcoholism and/or the modification of its severity. This is supported through animal experimental and pharmacological data. We analyzed the DRD2 311Ser/Cys polymorphism in 312 German alcoholics and 131 ethnically matched controls to investigate the association of genetic DRD2 variants with alcoholism or clinical characteristics of homogeneous subgroups of alcoholics. We observed no association between the 311Cys variant and alcoholism, and none of the clinical characteristics evaluated was significantly associated with 311Cys. The allele frequencies of the 311Cys variant were 0.026 and 0.031 in the alcoholics and controls, respectively. These are the highest reported 311Cys frequencies in Caucasians. The DRD2 TaqI A1/A2 restriction fragment length polymorphism was analyzed simultaneously in our samples. In most cases, the 311Cys allele is associated with the TaqI A2-allele. Data do not suggest a clinical relevance of the 311Cys variant in alcoholism. However, the relevance of this variant in other diseases or the existence of other DRD2 variants with altered receptor function or expression cannot be excluded. PMID- 8727250 TI - Human hepatic alcohol dehydrogenase and human erythrocyte catalase do not metabolize the cytochrome P-4502E1 substrate, chlorzoxazone. AB - Studies of cytochrome P-4502E1 (CYP2E1)-mediated oxidation of ethanol have been hampered by the lack of a suitable probe for in vivo human studies. Chlorzoxazone, a prescribed skeletal muscle relaxant, is metabolized to 6 hydroxychlorzoxazone by CYP2E1 and has been advocated as a specific probe of this enzyme on the basis of microsomal studies. The applications of this probe may include delineating the contribution of CYP2E1 to in vivo human ethanol metabolism. However, the activity of nonmicrosomal enzymes in metabolizing chlorzoxazone is unknown. Alcohol dehydrogenase (ADH), predominantly a hepatic cytosolic enzyme, may be more important than CYP2E1 in the oxidation of ethanol to acetaldehyde. The contribution of catalase in the in vivo oxidation of ethanol to acetaldehyde is controversial. To determine if either of these enzymes metabolizes chlorzoxazone and whether ethanol oxidation by either enzyme is inhibited by chlorzoxazone or its metabolite, multiple in vitro studies were performed. ADH enzyme kinetics were performed with human recombinant beta 1 beta 1 and beta 3 beta 3 ADH with ethanol and chlorzoxazone (0.5 to 2.5 mM). Neither ADH isoenzyme exhibited NAD(+) -dependent oxidation of chlorzoxazone, but displayed Michaelis-Menten kinetics for ethanol with K(m) values of 89 microM and 34 mM, for beta 1 beta 1 and beta 3 beta 3, respectively. Typical in vivo concentrations of chlorzoxazone and its metabolite, 6-hydroxychlorzoxazone, did not alter beta 1 beta 1 or beta 3 beta 3 ADH-mediated oxidation of ethanol to acetaldehyde. Studies of human hepatic nonmicrosomal enzyme activity were expanded to include all nonmicrosomal NAD(+) -dependent hepatic enzymes by starch gel electrophoresis assessment. Human hepatic enzymatic activity in the presence of chlorzoxazone was similar to that observed in the control sample (no added substrate), suggesting a lack of metabolism by NAD(+)-dependent enzymes. Similarly, human erythrocyte catalase, in the presence of a hydrogen peroxide generating system, did not metabolize chlorzoxazone. Furthermore, neither chlorzoxazone nor 6-hydroxychlorzoxazone altered the catalase-induced formation of acetaldehyde from ethanol. These data are consistent with chlorzoxazone as a specific probe of CYP2E1 that may be useful to alcohol researchers. PMID- 8727251 TI - Effect of chronic alcohol consumption on responses of cerebral arterioles. AB - The goal of this study was to determine whether chronic ingestion of alcohol alters dilatation of cerebral arterioles in response to agonists that produce activation of adenylate cyclase and activation of ATP-sensitive potassium channels. Rats were fed liquid diets with or without ethanol for 2 to 2.5 months. In vivo diameter of pial arterioles was measured in alcohol-fed and nonalcohol fed rats during superfusion with isoproterenol, forskolin, cromakalim, and nitroglycerin. Dilatation of pial arterioles in response to activation of adenylate cyclase via stimulation of beta-adrenergic receptors using isoproterenol was impaired in alcohol-fed rats. Isoproterenol (1.0 microM) dilated cerebral arterioles by 15 +/- 3% in nonalcohol-fed rats, but by only 7 +/ 2% in alcohol-fed rats. In contrast, dilatation of pial arterioles in response to forskolin was similar in nonalcohol-fed and alcohol-fed rats. Dilatation of pial arterioles in response to activation of ATP-sensitive potassium channels was impaired in alcohol-fed compared with nonalcohol-fed rats. Cromakalim (1.0 microM) dilated pial arterioles by 22 +/- 5% in nonalcohol-fed rats, but by only 5 +/- 2% in alcohol-fed rats (p < 0.05). In contrast, dilatation of pial arterioles in response to nitroglycerin was similar in alcohol and nonalcohol-fed rats. The findings of the present study suggest that chronic alcohol ingestion impairs dilatation of cerebral resistance arterioles in response to activation of adenylate cyclase via stimulation of beta-adrenergic receptors and in response to activation of ATP-sensitive potassium channels. We suggest that impaired vasodilator mechanisms during chronic alcohol consumption may have important implications for the pathogenesis of cerebrovascular abnormalities observed during chronic alcoholism. PMID- 8727252 TI - Quantitative trait locus analyses of sleep-times induced by sedative-hypnotics in LSXSS recombinant inbred strains of mice. AB - The long-sleep (LS) and short-sleep (SS) selected lines of mice show highly significant differences in sleep-time for many sedative-hypnotic drugs, and the quantitative genetic nature of these differences has been well-established. Using an interval-mapping approach, quantitative trait locus (QTL) analyses of LSXSS recombinant inbred (RI) strains have been applied to sleep-time responses for various classes of sedative-hypnotic drugs: alcohols (ethanol, n-propanol, and n butanol), the atypical anesthetic chloral hydrate, barbiturates (pentobarbital and secobarbital), and benzodiazepines (chlordiazepoxide and flurazepam). Several provisional QTLs were mapped to similar locations within and between drug classes, suggesting that some common loci are involved in sleep-times elicited by these drugs. Consistent with correlations of strain mean sleep-times between drugs tested in the LSXSS recombinant inbred strains, the number of provisional QTLs mapping to the locations of highest significance for ethanol decreases when the lipid solubility of a particular drug becomes less similar to that of ethanol. Provisional QTLs mapped for the benzodiazepines, however, revealed considerable overlap with those mapped for ethanol, although these drugs represented the most lipid-soluble category of sedative-hypnotics tested. Provisional QTLs for pentobarbital and secobarbital differed from most of those mapped for the alcohols, which supports the hypothesis that alcohols and barbiturates exert their effects mainly through different biological mechanisms in the LS and SS lines. Blood ethanol concentrations at regaining the righting reflex also mapped to several provisional QTLs corresponding to ethanol-induced sleep-times that support the contention that sleep-time is a reasonable index of the observed differences in central nervous system sensitivities to ethanol between LS and SS mice. PMID- 8727253 TI - Aldehyde dehydrogenases of the rat colon: comparison with other tissues of the alimentary tract and the liver. AB - Intracolonic bacteria have previously been shown to produce substantial amounts of acetaldehyde during ethanol oxidation, and it has been suggested that this acetaldehyde might be associated with alcohol-related colonic disorders, as well as other alcohol-induced organ injuries. The capacity of colonic mucosa to remove this bacterial acetaldehyde by aldehyde dehydrogenase (ALDH) is, however, poorly known. We therefore measured ALDH activities and determined ALDH isoenzyme profiles from different subcellular fractions of rat colonic mucosa. For comparison, hepatic, gastric, and small intestinal samples were studied similarly. Alcohol dehydrogenase (ADH) activities were also measured from all of these tissues. Rat colonic mucosa was found to possess detectable amounts of ALDH activity with both micromolar and millimolar acetaldehyde concentrations and in all subcellular fractions. The ALDH activities of colonic mucosa were, however, generally low when compared with the liver and stomach, and they also tended to be lower than in small intestine. Mitochondrial low K(m) ALDH2 and cytosolic ALDH with low K(m) for acetaldehyde were expressed in the colonic mucosa, whereas some cytosolic high K(m) isoenzymes found in the small intestine and stomach were not detectable in colonic samples. Cytosolic ADH activity corresponded well to ALDH activity in different tissues: in colonic mucosa, it was approximately 6 times lower than in the liver and about one-half of gastric ADH activity. ALDH activity of the colonic mucosa should, thus, be sufficient for the removal of acetaldehyde produced by colonic mucosal ADH during ethanol oxidation. It may, however, be insufficient for the removal of the acetaldehyde produced by intracolonic bacteria. This may lead to the accumulation of acetaldehyde in the colon and colonic mucosa after ingestion of ethanol that might, at least after chronic heavy alcohol consumption, contribute to the development of alcohol-related colonic morbidity, diarrhea, and cancer. PMID- 8727254 TI - Ethanol-induced alterations in the posttranslational processing, but not secretion of luteinizing hormone-releasing hormone in vitro. AB - The effects of ethanol (EtOH) on the male hypothalamic pituitary reproductive axis are multiple and varied. Although direct gonadal toxicity has been reported, hypothalamic-pituitary perturbations have also been noted. The difficulty of sampling the hypothalamus has made direct investigation of EtOH-induced alterations on luteinizing hormone-releasing hormone (LHRH) fraught with interpretation problems. To circumvent this, we have conducted a series of experiments exploring the effect of 200 mg% EtOH in vitro on GT1-7 cells, a newly developed LHRH secreting neural cell line. Cell lines were treated with EtOH containing or EtOH-free media for 2, 6, 24, or 48 hr. EtOH caused no significant change in LHRH secretion at any time point, although there was a trend to increased secretion after 2 hr EtOH exposure when compared with control. Significantly increased total (i.e., cellular plus secreted) pro-LHRH coupled with significantly reduced cellular LHRH after 6 hr only of EtOH exposure suggested that EtOH caused a transient decrease in processing from bioinactive pro-LHRH to bioactive LHRH. However, even at this time point, LHRH secretion from these EtOH-exposed cells was no different than from control cells. Steady-state LHRH mRNA levels were not changed by EtOH at any time point. These findings are concordant with previous in vitro data using hypothalamic tissue that has similarly demonstrated no effect of EtOH on LHRH secretion. Taken together with the in vivo demonstration that EtOH reduces hypothalamic-pituitary portal blood levels of LHRH, these data indicate that EtOH exerts its effect either at an extrahypothalamic locus and/or on non-LHRH-producing cells within the hypothalamus. PMID- 8727255 TI - Effect of bestatin, an aminopeptidase inhibitor, on alcohol intake in alcohol preferring P rats. AB - Bestatin is an aminopeptidase and enkephalinase inhibitor that elevates the levels of angiotensin II and angiotensin III. Although it has been used for years in the treatment of various forms of cancer, its application as an antialcohol drug has not been explored, despite its ability to stimulate angiotensin activity. The present study assesses the ability of bestatin to reduce chronic alcohol consumption in the genetically selected high alcohol-consuming P rats. Two groups of P rats were given 24-hr access to food, water, and 10% (v/v) alcohol. After a baseline period, half the rats received the saline vehicle and the other half ascending doses of bestatin (2.5, 10, 20, and 30 mg/kg, bid). Each dose was administered for a minimum of 7 days. Bestatin had little effect on water intake or weight gain, but did produce a dose-dependent reduction in alcohol intake that averaged 33% and was sustained over the course of the 1-month period of administration. Years of experience with bestatin have shown it to be safe and free of major side effects. The present findings suggest that bestatin (Ubenimex) might also be useful in the treatment of alcohol abuse in humans. PMID- 8727256 TI - Effects of alcohols on murine preimplantation development: relationship to relative membrane disordering potency. AB - During in vitro culture of murine preimplantation embryos, we have observed that exposure to 0.1% ethanol induces an immediate increase in intracellular calcium levels and subsequently accelerates embryogenesis. If the observed effects of ethanol on developing embryos is mediated by its membrane disordering potency, we hypothesized that the relative membrane disordering potencies of related alcohols would correspondingly effect embryonic intracellular calcium levels and developmental rates. Two-cell embryos were exposed to 0.1% ethanol or 0.05 to 1.0% (w/v) n-butanol, n-propanol, isopropanol, 1,2-propanediol, glycerol, or methanol for 24 hr at 37 degrees C, and development to the blastocyst stage was monitored after 5 days. n-Butanol, n-propanol, isopropanol, and methanol treatment caused a dose-dependent inhibition (p < 0.01) of development to the blastocyst stage, whereas 1,2-propanediol or glycerol neither accelerated nor inhibited development. In a second experiment, 8-cell morulae were treated with 1,2-propanediol or glycerol, and cavitation rates were examined. There was no significant difference from control embryos in the onset of cavitation or the blastocoel expansion rate of 1,2-propanediol- or glycerol-exposed embryos, whereas exposure to 0.1% ethanol accelerate cavitation (p > 0.05). In a third experiment, morulae were exposed to 0.1% or 1.0% of each alcohol and were monitored for changes in intracellular calcium levels using the fluorescent indicator, fluo-3-acetoxymethyl ester. There was an immediate increase in intracellular calcium levels when morulae were treated with 1.0% ethanol or n butanol, but only ethanol induced an increase (p < 0.05) in the level of intracellular calcium at 0.1%. These data suggest that ethanol is unique in its ability to accelerate embryogenesis and that the membrane disordering potency of ethanol does not directly underlie its effects on intracellular calcium release and the acceleration of preimplantation development. PMID- 8727257 TI - Ethanol inhibits human osteoblastic cell proliferation. AB - The habitual consumption of alcoholic beverages is clearly associated with low bone mass and an increased prevalence of skeletal fractures. Microscopic analysis of skeletal tissue from alcoholic patients reveals reduced osteoblast number and suppressed bone formation activity with a relative sparing of resorptive indices. The decreased number of osteoblasts observed in alcoholic subjects results from either impaired proliferation or accelerated senescence. Polyamines and ornithine decarboxylase (ODC), the rate-limiting enzyme for polyamine synthesis, are essential for cell proliferation in a variety of cell types. To determine if the adverse effect of ethanol on osteoblast number involves modulation of polyamine biosynthesis, we examined the effect of ethanol on parameters of cell growth and ODC activity in a human osteoblast-like osteosarcoma cell line (TE-85). Ethanol markedly impaired DNA synthesis and cell proliferation in a dose-dependent fashion, but alkaline phosphatase activity (a marker of differentiated osteoblast function) remained intact, and accelerated apoptosis was not evident. Thus, the reduced osteoblastic cell number was a result of a direct effect on proliferative processes rather than a nonspecific toxic effect of ethanol to accelerate cell death. Induction of ODC activity was impaired in ethanol-exposed cell cultures in a dose-dependent fashion that paralleled the antiproliferative effects. Finally, supplemental polyamine administration substantially improved DNA synthesis in ethanol-exposed UMR 106-01 cell cultures. These data confirm a direct inhibitory effect of ethanol on osteoblast proliferation without overt cellular toxicity that may, in part, explain the reduced bone mass observed in those who consume excessive amounts of alcohol. PMID- 8727258 TI - Effects of chronic ethanol administration on the endocytosis of cytokines by rat hepatocytes. AB - The effects of chronic ethanol administration on the endocytosis of three representative cytokines were investigated in isolated rat hepatocytes. When hepatocytes were isolated from rats that were fed an ethanol liquid diet for 12 to 13 weeks, these cells exhibited a decreased ability to internalize and degrade transforming growth factor-alpha, tumor necrosis factor-alpha and interleukin-6, compared with hepatocytes from the pair-fed controls. This impaired endocytosis of all three cytokines was accompanied by significant decreases in the amount of hepatocyte surface-bound cytokine. Changes in cytokine binding to surface receptors and reduced rates of receptor-cytokine complex internalization into the cells seem to be major contributors to defective endocytosis in hepatocytes from the ethanol-fed rats. Impaired hepatocyte endocytosis could lead to altered steady-state levels of cytokines in the liver and modified physiological responses to cytokines. These changes could affect homeostasis among the various cell types in the liver and could contribute to liver dysfunction and injury. PMID- 8727259 TI - Naltrexone persistently reduces rats' intake of a palatable alcoholic beverage. AB - Rats were given 30 days of opportunity to take a sweetened alcoholic beverage and water for 2 hr/day. At first, they took little alcohol, but subsequently took, on average, 2.3 g/kg of alcohol/daily session. They also took sufficient water, during the 2-hr period, to maintain their health and to steadily gain weight. At the end of the 30 days, they were divided into four groups so that their intakes of alcohol were similar. All groups continued on the daily regimen, but each group received different injections. One group received placebos, whereas the other two groups received either 5.0 or 10.0 mg/kg, respectively, of naltrexone daily, 30 min before the drinking session. The fourth group received 5.0 mg/kg of naltrexone 12.5 hr before the session and another 5.0 mg/kg 30 min before the session. This regimen of dosing and daily opportunities to drink continued for 30 days. With the end of injections, subjects continued on the regimen for another 5 days. Naltrexone, dose-relatedly, reduced rats' intake of alcoholic beverage. Furthermore, with respect to reducing intake of alcohol, no tolerance or refractoriness were observed across the 30 days of dosing. Within a couple of days after dosing, levels of intake returned to predosing levels. PMID- 8727260 TI - Zonal differences in ethanol-induced impairments in fluid-phase endocytosis in rat hepatocytes. AB - We have previously shown that ethanol administration impairs the processes of fluid-phase endocytosis (FPE) and receptor-mediated endocytosis (RME) in isolated rat hepatocytes after as early as 1 week of ethanol administration. The defects in RME were most prominent in the perivenule (PV) region of the liver lobule, the area wherein alcoholic liver disease has been shown to start and predominate. We undertook the present study to see if changes in FPE were likewise more apparent in the PV versus the periportal (PP) region of the liver. For these studies, we fed male Sprague-Dawley rats with an ethanol-supplemented liquid diet or an isocaloric control diet for 1 or 5 weeks. PV and PP hepatocytes were isolated using a digitonin-collagenase perfusion method. Internalization and efflux of the marker dye, Lucifer Yellow, was then examined in the cell populations. After as early as 1 week of feeding, cells from the PV region in ethanol-fed animals showed dramatic impairments in the net internalization of dye, compared with PV cells from the pair-fed controls, and these changes persisted throughout the 5 week feeding period. In contrast, internalization of Lucifer Yellow in cells from the PP region of the liver were not different between control and ethanol animals. Because net internalization represents the balance between uptake into the cells versus efflux from the cells, we examined these components individually. Early uptake of the dye into the cell was not altered by ethanol treatment. The decreased net internalization seemed to be caused by enhanced efflux of the dye, which was significantly increased in PV cells, compared with the same cell type in control animals. Cells from the PP region of the ethanol fed animals did not exhibit altered efflux after either 1 or 5 weeks of feeding. These results indicate that ethanol-induced impairments in FPE are more dramatic in the PV region of the liver, and these impairments seem to result from an ethanol-induced enhancement of efflux. PMID- 8727261 TI - S-methyl-N,N-diethylthiocarbamate sulfoxide and S-methyl-N,N-diethylthiocarbamate sulfone, two candidates for the active metabolite of disulfiram. AB - The mechanism of action of disulfiram involves inhibition of hepatic aldehyde dehydrogenase (ALDH). Although disulfiram inhibits ALDH in vitro, it is believed that the drug is too short-lived in vivo to inhibit the enzyme directly. The ultimate inhibitor is thought to be a metabolite of disulfiram. In this study, we examined the effects of S-methyl-N,N-diethylthiocarbamate (MeDTC) sulfoxide and S methyl-N,N-diethylthiocarbamate sulfone (confirmed and proposed metabolites of disulfiram, respectively) on rat liver mitochondrial low K(m) ALDH. MeDTC sulfoxide and MeDTC sulfone, in 10-min incubations with detergent-solubilized mitochondria, inhibited ALDH activity with an IC50 (mean +/- SD) of 0.93 +/- 0.04 and 0.53 +/- 0.11 microM, respectively, compared with 7.4 +/- 1.0 microM for the parent drug disulfiram. Inhibition by MeDTC sulfone and MeDTC sulfoxide, both at 0.6 microM, was time-dependent, following apparent pseudo-first-order kinetics with a t1/2 of inactivation of 3.5 and 8.8 min, respectively. Dilution of ALDH inhibited by either sulfoxide or sulfone did not restore activity, an indication of irreversible inhibition. Addition of glutathione (50 to 1000 microM) to ALDH before the inhibitors did not alter the inhibition by MeDTC sulfoxide. In contrast, the inhibition by MeDTC sulfone was decreased > 10-fold (IC50 = 6.3 microM) by 50 microM of glutathione and almost completely abolished by 500 microM of glutathione. The cofactor NAD, in a concentration-dependent manner, protected ALDH from inhibition by MeDTC sulfoxide and MeDTC sulfone. In incubations with intact mitochondria, the potency of the two compounds was reversed (IC50 of 9.2 +/- 3.6 and 0.95 +/- 0.30 microM for the MeDTC sulfone and sulfoxide, respectively). Our results suggest that MeDTC sulfone is highly reactive with normal cellular constituents (e.g., glutathione), which may protect ALDH from inhibition, unless this inhibitor is formed very near the target enzyme. In contrast, MeDTC sulfoxide is a better candidate for the ultimate active metabolite of disulfiram, because it is more likely to be sufficiently stable to diffuse from a distant site of formation, such as the endoplasmic reticulum, penetrate the mitochondria, and react with ALDH located in the mitochondrial matrix. PMID- 8727262 TI - Quantitative autoradiography using selective radioligands for central and peripheral-type benzodiazepine receptors in experimental Wernicke's encephalopathy: implications for positron emission tomography imaging. AB - Wernicke's encephalopathy (WE) is difficult to diagnose during life, with up to 80% of cases being missed by routine neurological evaluation in both alcoholics and AIDS patients. Therefore, there is a need for noninvasive diagnostic procedures. Using the pyrithiamine-treated rat, an animal model of WE, we have studied, using radioligands for central (neuronal) and "peripheral-type" (glial) benzodiazepine receptors, the regional distribution of changes in the densities of these receptors in relation to the degree of reactive gliosis accompanying neuronal loss. Histological studies revealed neuronal loss in selective regions, including the thalamus, inferior colliculus, inferior olivary nucleus, and mammillary body. Autoradiographic studies demonstrated increases in densities of [3H]PK11195 binding sites that closely paralleled the topographic distribution of neuronal cell loss and reactive gliosis. In contrast, [3H]Ro15-178 showed poor spatial correlation, with the neuronal loss seen in pyrithiamine-induced thiamine deficiency. The positron emission tomography ligand [11C]PK11195 may be useful for the assessment of thiamine deficiency-induced brain damage in human alcoholics. PMID- 8727263 TI - Alternative routes for allergen-specific immunotherapy. AB - The alternative routes for allergen-specific immunotherapy (oral, sublingual, intranasal) have the overall aim of minimizing or avoiding the possible side effects caused by the injectory route, and of making the treatment more convenient and acceptable for the patients. The clinical efficacy and the safety of the alternative routes have been clearly demonstrated in many controlled studies for the most common inhalant allergens. The oral routes appear particularly suitable for children and patients with unsatisfactory compliance with the injectory route. On the other hand, nasal immunotherapy, because of its peculiar administration technique, requires a careful choice of well-trained adult patients. In conclusion, the good tolerability, safety and socioeconomic benefits strongly support the use of alternative routes as valid therapeutic options for the treatment of respiratory allergy. PMID- 8727264 TI - Specific IgE determination in the diagnosis of beta-lactam allergy. AB - Allergic reactions to beta-lactams are the most frequent adverse reactions to drugs, but conventional methods of detection of beta-lactam specific IgE detect only a low number of positivities. In this study we evaluated the diagnostic reliability of one of these methods (CAP FEIA, Pharmacia) which has been proven very useful in inhalant and food specific IgE determinations. We studied specific IgE to penicillin G, penicillin V, amoxicillin and ampicillin in 1078 patients who had been diagnosed with an adverse reaction to beta-lactams, and out of this group, in a selected subgroup of 149 patients with a very suggestive clinical history of beta-lactam anaphylactic reaction. Also, positivity of specific IgE was studied in a group of 58 patients with a clinical history of immediate hypersensitivity to beta-lactams and with positive skin tests, and positivity of skin tests was studied in a subgroup of 13 patients with positive specific IgE. To increase the sensitivity of IgE determinations, we concentrated 5-fold the sera from six selected patients. In the first group we found that penicillin G specific IgE was positive in 28% of the cases when considering a cutoff > or = class 1, and 6% when considering a cutoff > or = class 2. For penicillin V, positivities were 9% (> or = class 1) and 4% (> or = class 2). In the second group, we found 31.81% sensitivity and 88.57% relative specificity of CAP FEIA with respect to the skin test. In the group of 58 patients with a clinical history of immediate hypersensitivity to beta-lactams, we found a disagreement between skin testing and specific IgE determination in 28.2%, and a positive agreement in 9.4% of the cases. After sera concentration, there was a significant increase (p < 0.001), from 0.58 to 1.61 kU/l, in beta-lactam specific IgE. Our results suggest that skin testing offers greater reliability and sensitivity than beta-lactam specific IgE determination by CAP FEIA. PMID- 8727265 TI - Storage mite allergy in perennial rhinitis patients not sensitized to house dust mites. AB - Antigens from house dust mites are the most important indoor allergen involved in the immunopathogenesis of perennial allergic rhinitis and asthma. Storage mites have been identified in the dust of homes in many tropical and temperate climates. The role of these mites in allergic respiratory conditions needs to be clarified. We studied sensitivity to the storage mites Biomia tropicalis and Tyrophagus putrescentiae in patients previously considered non-allergic, based on negative reactions to skin tests with common inhalant allergens in our region. It was possible to verify that some of these patients were in fact sensitized exclusively to the storage mite allergens. PMID- 8727266 TI - Sequential determinations of Dermatophagoides spp. allergens in a tropical city. AB - The purpose of this study was to establish seasonal levels of Der p 1, Der f 1 and D. pteronyssinus allergens in mattress and floor dust samples in the homes of 20 mite-allergic asthmatic patients in Cartagena, Colombia. Dust samples were collected using a portable vacuum cleaner once monthly for the same 12 months in all houses. The highest Der p 1 levels in mattress samples were detected in August (geometric mean = 109.49 ng/g) and represented a 2.2-times increase from the lowest level observed in November. Der f 1 was detected in only 3 mattresses, ranging from 90 ng to 1.6 micrograms per gram of dust. The highest levels were detected in February and March. The highest level of D. pteronyssinus allergens in mattress dust samples, quantitated by RAST-inhibition, was 8, 994 AU/g (80% inhibition); the lowest level detected was 150 AU/g (5% inhibition). Allergen levels were significantly higher in mattresses than in floor samples (p < 0.001). Overall, a Spearman rank correlation coefficient of 0.48, p < 0.001, was obtained between Der p 1 and D. pteronyssinus allergens. There was a good correlation between mite allergens and absolute humidity (r = 0.8, p = 0.007). Der p 1, Der f 1 and D. pteronyssinus allergen levels show minimal variations in this tropical environment. Allergens derived from various mite species must be considered when assessing mite allergen exposure in these environments. PMID- 8727267 TI - Evaluation of the efficacy of oral cromolyn sodium or an oligoantigenic diet in children with atopic dermatitis: a multicenter study of 1085 patients. AB - One thousand eighty-five children with atopic dermatitis were enrolled in a multicenter study to evaluate the efficacy of 4 weeks of oral sodium cromoglycate or 4 weeks of a restricted diet. One thousand-eleven children (93%) concluded the study. At the end of the trial there was a significant improvement in skin lesions in the two groups: 61% of the patients in the sodium cromoglycate group and 69% in the restricted diet showed a significant improvement in atopic dermatitis. We concluded that, at least in our experimental design, both sodium cromoglycate and a restricted diet are equally effective in atopic dermatitis. PMID- 8727268 TI - Non-specific nasal provocation in children with chronic allergic rhinitis. AB - Twenty-three children with chronic allergic rhinitis (Group A) and eleven normal non-atopic subjects (group C) were submitted to non-specific nasal provocation tests with histamine (0.03; 0.06; 0.125; 0.25; 0.5; 1.0; 2.0 and 4.0 mg/ml), and one week later with methacholine (0.025; 0.1; 0.25, 0.5; 1.0; 2.5; 5.0 and 10.0 mg/ml). Measurements of total nasal resistance were performed by active anterior rhinomanometry (Berger, S.A.) and symptoms were recorded. Challenges were carried out in the morning with all children in an acclimatized room (25 degrees C/77 degrees F). Concentrations of the tested drugs were increasingly instilled, and after 5 min were followed by total nasal resistance, FEV, and FVC measurements. Considering as positive those tests in which total nasal resistance had a 100% increase, we observed that both histamine and methacholine caused an increase in total nasal resistance as instilled drug concentration rose; histamine provocations were significantly more positive among group A than group C patients (91% sensitivity, 80.8% positive predictive value). This was not observed with methacholine (55% sensitivity, 75% positive predictive value). In neither provocation was there correlation between the concentration that induced a positive response and symptoms. There were no changes in spirometric values during the tests. Nasal provocations with histamine and methacholine are safe and well tolerated. Histamine seems to be more adequate for differentiating children with allergic rhinitis from normal controls. PMID- 8727269 TI - Sensitivity to Cupressus: allergenic significance in Cordoba (Spain). AB - The detection of high levels of Cupressaceae pollen concentration in the air from January to April for several years in our area prompted analysis of the incidence and allergenic significance of sensitivity to this pollen. Furthermore, this is the highest winter-blooming taxa in the city of Cordoba. Skin prick tests were carried out over a one-year period on 1532 patients suffering from respiratory disorders (asthma and/or rhinoconjunctivitis). A total of 42 variables were studied in Cupressus-positive and Cupressus-negative patients; the data obtained were analyzed using a statistical software package. Sensitivity to Cupressus was found in 13% of all outpatients attending the unit, 18% of patients with respiratory disorders and 35% of patients with pollinosis. No significant differences were found between Cupressus-positive (C+) and Cupressus-negative (C ) patients with regard to mean age, sex, patient environment (i.e., rural, semi rural, urban), personal or family history of atopy, clinical symptoms or evolution after immunotherapy (which did not include this antigen). More C+ patients were found in the higher age brackets (over 25 years old; p < 0.05); C+ patients showed greater duration (p < 0.05) and slower development (p < 0.05) of symptoms, and were also found to be more sensitive to other pollens (p < 0.001). All the Cupressus-sensitive patients also reacted positively to Olea and Fraxinus, compared to 77% and 51% in the two Cupressus-negative groups. PMID- 8727270 TI - The influence of misoprostol (synthetic analogue of prostaglandin E1) on aspirin induced bronchoconstriction in aspirin-sensitive asthma. AB - It is believed that aspirin (ASA) and other nonsteroidal anti-inflammatory drugs elicit dyspnea in ASA-sensitive asthmatics by blocking cyclooxygenase. It is unclear whether this bronchospasm is due to the shunting of arachidonic acid into the lipoxygenase pathway or to the removal of a cyclooxygenase product which prevents bronchospasm. Diminished tissue concentration of PGE may cause bronchoconstriction. PGE also modulates mast cells, decreasing the release of anaphylaxis mediators. The authors investigated the influence of a synthetic analogue of PGE1-misoprostol (Cytotec, Searle)-on post-aspirin bronchoconstriction in seven ASA-sensitive asthmatics. On the first day, the effect of a placebo was studied. On the second day, the bronchodilatory effect of misoprostol (Cytotec, Searle) alone was examined. After a few days, a predetermined threshold dose of ASA was administered. Seven days later, at least 400 micrograms of misoprostol +200 micrograms 2 h later, together with a predetermined ASA dose, were administered. In all but one patient, the protective influence of misoprostol on ASA-induced bronchoconstriction was observed. The maximum drop in FEV1 (forced expiratory volume in one second) in % after ASA in each of the patients was 40, 25, 24, 33, 47 and 54, and after ASA with misoprostol 10, 9, 4, (+8), 10, (+2) and 45, respectively. Misoprostol given together with ASA attenuated aspirin-induced bronchoconstriction, reaching statistical significance at 3 and 3.5 h. It also diminished extrapulmonary symptoms. PMID- 8727271 TI - Methotrexate in the treatment of corticodependent asthmatic children. AB - Methotrexate has been used as an anti-inflammatory agent in chronic asthma. We evaluated the action of methotrexate in eight corticodependent severely asthmatic children (more than 10 mg of prednisone per day for at least one year). The patients (3 males and 5 females; aged 8 to 14 years) received a single weekly dose of 0.6 mg/kg methotrexate (maximum 25 mg) and folic acid (15 mg/day for 5 days in the week). The children were examined and had their pulmonary function test evaluated weekly. As the study progressed, the dose of prednisone was reduced and maintained till the next evaluation if the patient's symptoms were under control. After the 3rd month of treatment, we observed a significant reduction in the dose of prednisone and maintenance of the spirometric parameters. At the end of the trial, in 4 patients it was possible to reduce the basal prednisone dose 56% or more. In the remaining 4, one did not show any benefit and in the other 3, it was possible to obtain an average reduction of 40% of the basal prednisone dose. The total mean reduction was 55.9%. This oral corticoid reduction was not associated with clinical or pulmonary function deterioration, except in one patient. The patients were submitted to white blood cell count, hepatic transaminases, urine tests and other determinations at least once a month. There were no changes in biochemical tests. The side-effects were nausea, vomiting and abdominal pain. In conclusion, methotrexate given to severely corticodependent asthmatic children permitted a reduction in the daily intake of prednisone, reducing the severe side-effects of chronic corticotherapy. PMID- 8727272 TI - Fixed exanthema due to paracetamol. AB - Fixed exanthema represents 10% of the adverse reactions to medications. Up to 80% of the cases are caused by drugs. The lesions are frequently located on the genitals, perianal region, limbs and oral mucosa, in descending order of frequency. We describe the case of a 36-year-old man who developed fixed exanthema following administration of paracetamol. PMID- 8727273 TI - Theory and practice in speech-language pathology: a review of systemic principles. AB - Linear, cause-effect reasoning has dominated the training and clinical practice of speech-language pathology for many years. Recently, however, the roles of many speech-language clinicians and the contexts in which they provide their clinical services have been changing, and traditional ways of viewing clinical work no longer seem to apply. This article applies principles of systems theory to such changes and advocates an integration of these principles with those of traditional, linear views to achieve a better understanding of this evolution in the clinical practice of speech-language pathology. PMID- 8727274 TI - Systemic concepts in speech-language treatment. AB - While systemic concepts are well known in many professions, a systemic paradigm is relatively new to most speech-language pathologists. Five systems concepts that apply to current practices in speech-language pathology are described and illustrated. The concepts relate to the influences of context or environment, relationships, family structure, isomorphism, and the attribution of meaning. PMID- 8727275 TI - Use of the family to facilitate communicative changes in adults with neurological impairments. AB - Systemic concepts may be used to expand upon the traditional notion that family members influence the outcome of treatment services to adults with neurological communicative disorders. Several systemic concepts are discussed and illustrated as they are used with different clients having (1) aphasia, (2) right cerebral hemisphere dysfunction, and (3) progressive dementia. Discussion focuses on determining the unit of treatment with which to work, promoting and measuring positive communicative changes, and selecting effective intervention techniques. PMID- 8727277 TI - On becoming a team: a view from the field. AB - Early intervention services have expanded the concept of team participation for speech-language pathologists. Unlike traditional teams that grew out of the individual, direct service model of treatment, early intervention teams include the child's family along with professionals. Family members are invited to participate in assessing and treating their child, and the child is seen from a slightly different perspective by each team participant. Different viewpoints must be integrated and used to the benefit of the child. In this way, treatment options are expanded. Further, professionals may release their roles to other team members as well as learn new skills from other team members. The process of team development is illustrated through the experiences of one family-centered early intervention team. PMID- 8727276 TI - Collaborative consultation: a systemic activity. AB - In a collaborative consultation model of speech-language service delivery, the treatment system is expanded beyond the traditional client-centered dyad. Further, services are provided in the context of the classroom rather than in the noncontextual environment of a therapy room, and treatment goals relate to aspects of the school, such as curriculum, classroom behavior, and social interactions with peers. The use of the model is illustrated with a child having a cognitive impairment. PMID- 8727278 TI - Moving toward a systemic training model in a university program. AB - While the medical model continues to provide a basis for clinical service and the preparation of future speech-language pathologists, the instructor who teaches and uses systemic principles enriches the world view of students and prepares them for new roles being assumed by speech-language pathologists. Experiences of the author are described as she became aware of systemic principles and began teaching them to undergraduate and graduate students. Two formal strategies are discussed. These are linked to a Triadic Interaction Model and a Family-Based Treatment model. Students commented favorably on their experiences as they attempted to learn systemic principles and put them into practice in their clinical practicum. PMID- 8727279 TI - Working within systems that don't ... or at least not the way I thought they would! AB - Each culture has its own systems, values, structures, and rules which differ from those of other cultures. To function effectively in any culture, one has to gain an appreciation of its values and rules, an understanding of how it is organized, and knowledge of the people who are in charge of its systems. This is true, also, of clinical work with people of different cultures. This article is a personal journal which describes one speech-pathologist's encounters with ethnic and governmental cultures, what she learned, and how she grew. PMID- 8727280 TI - Keraunopathology. An analysis of 45 fatalities. AB - An analysis of 45 victims of fatal lighting strike revealed the incident occurred most frequently in the early afternoon during midsummer in a field of one form or another. The scene of death, damage to clothing, and alterations of metallic objects on the victim are described. A terminal cardiac rhythm of ventricular fibrilation was recorded in half for whom data were available, and asystole was found in 40%. All but four had cutaneous injuries, and nearly one-third had pathognomonic patterns of erythematous arborization. When examined, tympanic membranes were found to be ruptured in > 80%. This study also revealed that craniocerebral injury and cardiac contusion can be serious direct consequences of lightning strike. A correlative approach to the investigation and autopsy of lightning victims is suggested. PMID- 8727281 TI - Lichtenberg figures. AB - The majority of cutaneous injuries seen in victims struck by lightning are superficial and heal without sequelae. Common cutaneous findings associated with lightning strike include punctate full-thickness burns, linear charring, and contact burns from overlying metal objects. Branching or ferning marks are an uncommon and unusual cutaneous manifestation. These ferning patterns are called Lichtenberg figures. Similar-appearing electrical phenomena were first noted by an 18th-century physicist of the same name. A case report with review of the literature is presented. The histopathologic features of the figures are described for the first time. PMID- 8727282 TI - The pathological approach to sudden infant death--consensus or confusion? Recommendations from the Second SIDS Global Strategy Meeting, Stavangar, Norway, August 1994, and the Third Australasian SIDS Global Strategy Meeting, Gold Coast, Australia, May 1995. PMID- 8727283 TI - Heat-related deaths in Philadelphia--1993. AB - A study of heat-related deaths associated with the 1993 heat wave in Philadelphia, Pennsylvania, was conducted. Most of these deaths were in the susceptible elderly with preexisting natural diseases who lived alone without air conditioning in upstairs bedrooms with windows shut, thus creating an even hotter environment. These excessive deaths under such conditions did not meet the standard clinical criteria for hyperthermia because of varying postmortem intervals. Therefore, the authors stress the utility of a postmortem definition of heat-related death to better define the magnitude of health risk posed by hot weather and warn public health and other agencies to take preventative measures. PMID- 8727284 TI - Gastric contamination of postmortem blood samples during blind-stick sample collection. AB - A blood sample drawn by a blind stick through the chest was collected for toxicologic examination in a suspected natural death. Drug screen results on the blood sample indicated a combined overdose of alcohol and amitriptyline. An autopsy was later performed and a peripheral blood sample was drawn. A drug screen of this sample showed the presence of therapeutic quantities of amitriptyline and a trace of alcohol. Cytologic examination of the blind-stick sample showed the presence of columnar cells and food debris, confirming that the sample was contaminated either by stomach contents or gastric aspirate during the collection procedure. We strongly encourage all agencies to refrain from the use of blind-stick sample collection for toxicologic specimens. PMID- 8727285 TI - Medicolegal death investigator preemployment test development. AB - The medicolegal death investigator (MDI) represents an essential component in the multidisciplinary investigation of sudden and unexpected deaths. Presently there exists no specific program for the job training evaluation procedure for new (MDI) employees. We discuss the development of a medicolegal death investigator preemployment training tool, which consists of a training and evaluation program containing a list of tasks and task steps. After the investigator demonstrates the ability to perform individual tasks independently, the trainee will be objectively evaluated by the completion of a written test specifically designed to cover the task items. The preemployment test has been developed by those professionals best able to determine the job requirements, the death investigators. The specific outcomes of the process also include further definition of the profession, preemployment testing, policy and procedure development, posttraining evaluation, identification of training needs, curriculum design and further professional development, and identification of the profession of death investigator. PMID- 8727286 TI - Carboxyhemoglobin levels in a series of automobile fires. Death due to crash or fire? AB - The determination of death by trauma versus fire can be of major consideration, especially in civil product liability litigation. Blood carboxyhemoglobin levels can be instrumental in that differentiation. Twenty-eight fatalities involving fire in automobiles were reviewed. All subjects displayed some degree of body burn, and in 25 severe charring and/or incineration was present at autopsy. In only one case was there a history of explosion or flash fire. Carboxyhemoglobin levels varied from 92% to values of < 10%. In seven cases no collision occurred. In six of these subjects COHb values were > or = 47%. In all 16 cases with carboxyhemoglobin levels of < or = 10% a collision occurred. In 12 of 16 of these subjects, blunt force injury sufficient to cause death was discovered. Data presented in this article indicate that a carboxyhemoglobin level of > 30% strongly suggests inhalation of combustion products as the cause of death. In contrast, a level of < 20% should prompt a search for other causes. PMID- 8727287 TI - Wounding effects of the Winchester Black Talon bullet. AB - Winchester Ammunition has developed a bullet that was specifically designed to perform better than other hollow-point missiles. The resulting cartridge, the Black Talon bullet offers a low-velocity missile that retains its weight, penetrates even when interrupted by intervening objects, and remains capable of expansion in soft tissue. The bullet causes extensive tissue damage and presents a hazard to those responsible for handling it after expansion. PMID- 8727288 TI - Metallic dissolution of a civil war bullet embedded in a sternum. AB - The contemporary trend of converting departments of anatomy into departments of cell biology has brought with it the task of examining archive collections and storage facilities to figure out how to best utilize the available space. During one such inspection at the University of Louisville School of Medicine, a human sternum containing a dull metal projectile was uncovered. The projectile was easy to characterize as a bullet that had been deeply embedded in the bone. Less clear, however, were the circumstances detailing how the bullet had become lodged in the sternum, or how long the sternum might have been in storage at the University of Louisville. Radiographs of the sternum revealed a halo of surrounding density that dissipated in intensity from the margins of the bullet. Our initial hypothesis was that lead had been leached from the bullet into the bone matrix. To better assess what in fact contributed to this density, the sternum and the bullet were analyzed by energy-dispersive spectroscopy (EDS) to determine their elemental composition. That the bullet was composed of lead and aluminum was not surprising, but the extent to which the presence and dissolution of the bullet had affected the composition of the bony sternum was not entirely expected. The contribution of metal ions from the bullet to the inorganic matrix of bone was most notable for aluminum but nearly negligible for lead. This finding confirmed that bone affinity for metals is dependent upon the metal and supports previous reports that have suggested that lead is released from bone as soluble blood product during bony remodeling, whereas aluminum results in a significant elevation of bone density. PMID- 8727289 TI - The past and present legal weight of bite marks as evidence. AB - Bite mark evidence, which is most often associated with violent crimes, is legally accepted and admissible in a court of law. However, current legal attacks are underway against the admission of such evidence. Indeed, over the last 20 years, bite mark forensics have sustained a multitude of legal challenges, some concerning the constitutional rights of the accused and others the scientific acceptance of such an approach. Bite marks are considered hard evidence and are highly persuasive to juries who weigh the evidence. PMID- 8727290 TI - Left and right ventricular dysplasia and Uhl's anomaly. A case report. AB - Myocardial dysplasia involves a quasi-complete or localized absence of myocardial fibers in the right or left ventricle, with corresponding partial or complete filling by fibroadipose tissue. This disorder, in its full form called "Uhl's anomaly", was first described in 1952 as an anomaly of the heart conferring a parchment-like appearance to its walls. Incomplete forms of the disorder fall into the category of right or left arrhythmogenic ventricular dysplasia. Although approximately 50 cases of right ventricular dysplasia have been documented, only two reports concern left ventricular dysplasia. Having encountered the latter anomaly, we first present the case and our observations, and then review the etiology, pathogenesis, clinical aspects, and evolution of this disorder. PMID- 8727291 TI - Massive pulmonary tumor emboli in a sarcoma. An unusual cause of sudden death. AB - Although metastatic spread of tumor to the lungs is common, massive pulmonary tumor emboli are very unusual. Most tumor emboli originate from epithelial derived tumors. Only a few cases of pulmonary embolism from sarcomata have been reported. We herein describe the case of a 36-year-old woman who died suddenly due to massive pulmonary tumor emboli from a retroperitoneal malignant mesenchymoma. We believe this is the first case report of a mesenchymoma causing sudden death due to massive tumor embolism. PMID- 8727292 TI - Portal vein embolization following shotgun-pellet injuries to the abdomen. AB - Multiple missile emboli are rare. The most likely setting for their occurrence is multiple penetrating injuries from birdshot. Such is this case of this 26-year old man with birdshot injuries to the torso penetrating the portal vein and embolizing to the liver, producing infarcts and death. PMID- 8727293 TI - Cardiac muscle lesions associated with chronic administration of methamphetamine in rats. AB - Cardiovascular complications associated with methamphetamine abuse have increasingly been reported. However, chronic cardiotoxicity of methamphetamine is not experimentally well documented. In this study, methamphetamine (1 mg/kg/day) was subcutaneously injected into 5-week-old male Wistar Kyoto rats (n = 30). Age- and sex-matched Wistar Kyoto rats served as controls (n = 30). After 14 and 56 days, hearts were examined by light and electron microscopy. Foci of myocytic degeneration and necrosis appeared in the sub-endocardial areas on day 14 of methamphetamine exposure. Myocytic degeneration and necrosis became more extensive on day 56. At this stage, myocytolysis, contraction bands, atrophied myocytes, and spotty fibrosis were patchily distributed throughout the myocardium in most of rats treated with methamphetamine. The accompanying ultrastructural features included marked degeneration of cardiac mitochondria with fractured and disrupted cristae, hypercontraction of myofibrils, and loss of myofilament. In contrast, cardiac myocyte lesions were not observed in control rats. These myocardial lesions in rats treated with methamphetamine for 56 days resemble the cardiomyopathy associated with methamphetamine abuse in humans. PMID- 8727294 TI - Fatal posture- and heroin-related intestinal infarction and leg muscle necrosis after snorting heroin. A case report. AB - We present a case of a male patient who spent several hours sitting with legs crossed and upper body bent forward after nasal insufflation of heroin (snorting). Death occurred 3 days later as a result of leg muscle necrosis and intestinal ileus with infarction. The autopsy findings, including microthrombi in small mesenteric vessels and necrosis of the leg muscles, were attributed to vascular compression due to the man's position and to metabolic processes resulting from drug ingestion. PMID- 8727295 TI - Complete transection of the trunk of passengers in car accidents. AB - Traumatic amputation of extremities and complete severance of the trunk have been reported in extra-urban collisions between passenger cars and pedestrians at collision speeds of > 80-100 km/h (50-62 mi/h). In car passengers, such extreme types of injuries are very rare. Two cases are presented in which the cars had a lateral collision with road trees near the right B column (column between front and back door), as a result of which the car was torn into two parts just in front of the rear axle near the back seats. Under these circumstances, the trunk of the back passenger on the side of the collision was completely severed. Both accidents happened in left-hand bends and the speed of collisions amounted to 120 and 180 km/h (74 and 111 mi/h), respectively. All passengers were flung out of the cars. PMID- 8727296 TI - Author's reply. PMID- 8727297 TI - The role of pleural effusion in drowning. PMID- 8727298 TI - Purging: elimination of malignant cells from autologous blood or marrow transplants. PMID- 8727299 TI - Bone marrow transplantation from unrelated donors. AB - Unrelated donor bone marrow transplants provide the alternative to allogeneic transplants for patients lacking an HLA-matched related family member donor. Because the donor and recipient have greater immunogenetic disparity than related donor transplants, these transplants are associated with a higher rate of immunologic complications. This disparity has also translated into a lower risk of relapse relative to sibling donor transplants, enabling this treatment to offer a cure to advanced-disease patients. PMID- 8727300 TI - The graft-versus-leukemia effect. PMID- 8727301 TI - Donor lymphocyte infusions. AB - Donor lymphocyte infusions for treatment of relapse after allogeneic bone marrow or stem cell transplantation is being used with increasing frequency. Study of this form of adoptive immunotherapy will shed light on different aspects of cell mediated cytotoxicity such as antigen presentation, processing, immune recognition, lymphocyte subsets involved, and mechanism of cell death. Donor lymphocyte infusions are extremely effective for cytogenetic relapses or chronic phase relapses of chronic myelogenous leukemia, but are less effective in acute leukemias or other disorders. Donor lymphocyte infusions are associated with a significant risk of morbidity and mortality due to graft-versus-host disease and pancytopenia. Lower cell doses, earlier infusions, and selective depletion of CD8+ lymphocytes have been proposed as methods of diminishing these toxicities. Current research is focusing on methods of making donor lymphocyte infusions more effective in the nonchronic myelogenous leukemia setting, and decreasing their toxicity without losing their clinical efficacy in the treatment of relapsed chronic myelogenous leukemia. PMID- 8727302 TI - Preoperative chemotherapy for stage IIIa non-small cell lung cancer. AB - About 20% of patients with non-small cell lung cancer are stage IIIa at diagnosis. The treatment of stage IIIa tumors has been discouraging. Long-term disease control and cure rates with a single modality approach with surgery or radiotherapy have been poor; this is particularly so for N2 tumors, which account for the majority of stage IIIa disease. In the past decade there has been interest in multimodality treatment of stage IIIa non-small cell lung cancer using preoperative induction chemotherapy with or without radiotherapy followed by surgery. Several phase II studies and two small phase III trials have shown that a strategy of induction chemotherapy with or without radiotherapy is feasible and probably does offer some survival advantage over surgery or radiotherapy alone. The next issue to be tackled is to determine whether the results achieved with induction chemoradiation followed by surgery are equivalent to those of chemoradiation without surgery. A phase III intergroup trial is underway to answer this question. PMID- 8727303 TI - Thymic neoplasms. AB - Thymic carcinomas and thymomas are neoplasms of the epithelial cells of the thymus. Extensive study of the histopathology of these tumors has provided insight into the clinical behavior of various pathological subtypes. Establishment of a uniform staging system has facilitated the prospective study of thymic tumors, and it has helped to provide the necessary framework for international cooperation in the study of these diseases. This review explains the pathological nomenclature used in the classification of these tumors, examines the prospective clinical trials that have been completed to date, and discusses the interesting paraneoplastic phenomena associated with thymomas. PMID- 8727304 TI - Treatment of metastatic non-small cell lung cancer. AB - Although the overall survival rates for patients with metastatic non-small cell lung cancer have not changed in the past two decades, meta-analytic studies have confirmed that a modest increase in mean survival time can be gained with platinum-based combination chemotherapy. With appropriate selection of patients, chemotherapy will have symptomatic benefits in more than 60% of patients, and concerns regarding the costs of chemotherapy will be lessened by the observation that in some instances chemotherapy is less costly than best supportive care. Until the end of the 1980s, apart from the few active agents and their analogues no new drugs became available, but in the past 5 years several new agents have shown promising results and are now being included in combination programs. Large scale comparative studies, looking for the combination with the best therapeutic index, are awaited with great interest. The number of patients with non-small cell lung cancer is so great that even modest improvements in therapy will have a great impact on survival rates. PMID- 8727305 TI - Economic considerations in the care of lung cancer patients. AB - Lung cancer has been characterized as an expensive, futile, and self-induced illness. One of the most common questions pertaining to treatment is, "Is it worth it?" In the era of health care reform, attention has been directed toward common, high-cost illnesses that may benefit from closer examination of the clinical decisions that drive costs. This review explores the economic considerations of lung cancer treatment from the perspective of the patient, society, and those at risk for the costs of care. The concept of value is proposed as a frame-work to guide how lung cancer treatments should and should not be routinely used. Cost-effectiveness studies are highlighted that do not paint as dim a view of lung cancer therapy as may have been thought. However, it is clear that the 10 billion dollars spent yearly on lung cancer might be better used by limiting expenditures to the aspects of care that produce the best outcomes. This review includes comparisons of the cost-effectiveness of lung cancer care and treatments for other common cancers. It concludes with some strategies to use resources allocated to lung cancer more effectively. PMID- 8727306 TI - The current situation with regard to human melanoma and genetic inferences. AB - A melanoma predisposition gene has been identified. This gene, CDKN2, maps to chromosome 9p21-p22 and encodes p16, an inhibitor of cyclin-dependent kinases 4 and 6. CDKN2 has been found to be inactivated by homozygous deletion or intragenic mutation at high frequency in a diverse range of tumors and tumor cell lines, including those derived from melanomas. Now a number of CDKN2 mutations have been found in the germline of affected members of melanoma kindreds, and biochemical analysis of the mutant proteins has confirmed that they are functionally compromised. Unexpectedly, no germline CDKN2 mutations have been found in about half of the melanoma families that appear to be linked to 9p. Regulatory mutations outside of the coding region are being sought in these families. A number of other kindreds do not appear linked to 9p, hence the search continues for a second melanoma susceptibility gene. PMID- 8727307 TI - Pathology of malignant melanoma, including new markers and techniques in diagnosis and prognosis. AB - Although good prognostic markers are already available for patients with cutaneous melanoma, there still is a need for additional markers that are helpful to assess prognosis in individual patients. These so-called progression markers are likely to be found in molecules that play a role in the process of metastasis. Here, emphasis is put on the potential clinical implications of studies on the adhesion molecules from the integrin and CD44 families, proteases of the plasminogen activation system, and components involved in angiogenesis. Based on their differential expression in melanocytic tumor progression, several molecules of these categories appear promising for prognostic and diagnostic purposes. In this respect, the parallelism between key processes in the pathogenesis of metastasis and of angiogenesis is noteworthy. As technical developments in molecular pathology are relevant for diagnostic and prognostic purposes, some recent applications are discussed. An integrated molecular approach in a proper clinicopathologic context is advocated, including proper quality control measures. PMID- 8727308 TI - More rational and conservative surgical strategies for malignant melanoma using lymphatic mapping and sentinel node biopsy techniques. AB - Stimulated by novel lymphatic mapping techniques, the surgical care of the melanoma patient is becoming more conservative. Preoperative lymphoscintigraphy can identify all regional nodal basins at risk for metastatic disease, areas of "intransit" nodal collections, and the location and number of the primary draining nodes in relation to the rest of the nodes in the basin. Intraoperative mapping techniques, using a combination of a vital blue dye and a radioactive tracer, can then be used to harvest this first draining node, termed the sentinel node, for examination. It is clear that the most powerful prognostic factor for any solid tumor, including melanoma, is the presence or absence of regional lymph node disease. However, routine histologic examination may underestimate the number of patients with nodal disease. Serial sectioning and immunohistochemical staining, techniques that have been available for years, can increase the yield of positive dissections compared with routine histology. The selective approach to the nodal basin would allow the pathologist to be more detailed in examining the harvested sentinel node, providing more accurate staging information for melanoma. PMID- 8727309 TI - Sunburn, sunscreen, and melanoma. AB - This paper reviews the current epidemiologic and experimental evidence regarding the effect of sunburns on cutaneous malignant melanoma and the possible effectiveness of sunscreens in preventing those effects. Although there is growing agreement that sunlight exposure, particularly the ultraviolet wavelengths in solar radiation, contributes to the etiology of cutaneous malignant melanoma, there are at present insufficient data on the effective ultraviolet waveband and whether the use of sunscreens may be useful in preventing melanoma. The main obstacle in answering this question is that the exact role of sunlight in the pathogenesis of melanoma still remains undefined. However, new experimental animal models are now available that will assist in determining and defining the mechanism of initiation and promotion of melanoma by sunlight and, particularly, ultraviolet radiation in sunlight. While we await more definitive data, it is reasonable to recommend an overall "safe sun strategy" in which the use of sunscreens certainly must be accompanied by other protective measures of overall reduction of sunlight exposure. PMID- 8727311 TI - Transplantation. PMID- 8727310 TI - Interferons in melanoma. AB - The interferons are complex proteins that have been widely tested as therapy for neoplastic diseases. Interferon (IFN)-alpha has been the most extensively studied in melanoma. It produces responses in about 16% of metastatic melanomas, about one third of which are complete. It has been combined with chemotherapy and biologic therapy in an attempt to improve on this response rate, but despite encouraging reports from single-institution trials, firm evidence for improved efficacy is still forthcoming. Randomized trials of sequenced biochemotherapies should shed further light on this issue. The outlook in the adjuvant setting is much brighter with the recently reported positive results of the European Cooperative Oncology Group trial E1684 that showed improved overall and disease free survival for high-risk patients treated with IFN-alpha. The lack of efficacy of lower dosages and different schedules of administration argues for the use of maximally tolerated doses. IFN-gamma has proved disappointing in cases of both metastatic and high risk for relapse melanoma, but has been effective in locoregional disease, particularly in combination with tumor necrosis factor alpha and melphalan. Further research will need to focus on sequencing, improved methods of administration, and efforts to reduce toxicity. PMID- 8727312 TI - Lung and mediastinum. PMID- 8727313 TI - Melanoma and other skin neoplasms. PMID- 8727314 TI - Why are protein crystallographic R-values so high? PMID- 8727315 TI - Prediction of the secondary structure of HIV-1 gp120. AB - The secondary structure of HIV-1 gp120 was predicted using multiple alignment and a combination of two independent methods based on neural network and nearest neighbor algorithms. The methods agreed on the secondary structure for 80% of the residues in BH10 gp120. Six helices were predicted in HIV strain BH10 gp120, as well as in 27 other HIV-1 strains examined. Two helical segments were predicted in regions displaying profound sequence variation, one in a region suggested to be critical for CD4 binding. The predicted content of helix, beta-strand, and coil was consistent with estimates from Fourier transform infrared spectroscopy. The predicted secondary structure of gp120 compared well with data from NMR analysis of synthetic peptides from the V3 loop and the C4 region. As a first step towards modeling the tertiary structure of gp120, the predicted secondary structure may guide the design of future HIV subunit vaccine candidates. PMID- 8727316 TI - Molecular dynamics study of phospholipase A2 on a membrane surface. AB - The desolvation of lipid molecules in a complex of the enzyme human synovial phospholipase A2 with a lipid membrane is investigated as a mechanism that enhances the overall activity of the enzyme. For this purpose the interaction of the enzyme phospholipase A2 with a dilauryl-phosphatityl-ethanolamin (DLPE) membrane monolayer surface has been studied by means of molecular dynamics simulations. Two enzyme-membrane complexes, a loose and a tight complex, are considered. For comparison, simulations are also carried out for the enzyme in aqueous solution. The conformation, dynamics, and energetics of the three systems are compared, and the interactions between the protein and lipid molecules are analyzed. Free energies of solvation are calculated for the lipid molecules in the enzyme-membrane interface. Along with the calculated dielectric susceptibility at this interface, the results show the desolvation of lipids in a tightly bound, but not in a loosely bound protein-membrane complex. The desolvated lipids are found to interact mainly with hydrophobic protein residues, including Leu-2, Val-3, Ala-18, Leu-19, Phe-24, Val-31, and Phe-70. The results also explain why the turnover rate of phospholipase A2 complexed to a membrane is enhanced after a critical amount of negatively charged reaction product is accumulated. PMID- 8727317 TI - Constructing amino acid residue substitution classes maximally indicative of local protein structure. AB - Using an information theoretic formalism, we optimize classes of amino acid substitution to be maximally indicative of local protein structure. Our statistically-derived classes are loosely identifiable with the heuristic constructions found in previously published work. However, while these other methods provide a more rigid idealization of physicochemically constrained residue substitution, our classes provide substantially more structural information with many fewer parameters. Moreover, these substitution classes are consistent with the paradigmatic view of the sequence-to-structure relationship in globular proteins which holds that the three-dimensional architecture is predominantly determined by the arrangement of hydrophobic and polar side chains with weak constraints on the actual amino acid identities. More specific constraints are imposed on the placement of prolines, glycines, and the charged residues. These substitution classes have been used in highly accurate predictions of residue solvent accessibility. They could also be used in the identification of homologous proteins, the construction and refinement of multiple sequence alignments, and as a means of condensing and codifying the information in multiple sequence alignments for secondary structure prediction and tertiary fold recognition. PMID- 8727318 TI - Predicting solvent accessibility: higher accuracy using Bayesian statistics and optimized residue substitution classes. AB - We introduce a novel Bayesian probabilistic method for predicting the solvent accessibilities of amino acid residues in globular proteins. Using single sequence data, this method achieves prediction accuracies higher than previously published methods. Substantially improved predictions-comparable to the highest accuracies reported in the literature to date-are obtained by representing alignments of the example proteins and their homologs as strings of residue substitution classes, depending on the side chain types observed at each alignment position. These results demonstrate the applicability of this relatively simple Bayesian approach to structure prediction and illustrate the utility of the classification methodology previously developed to extract information from aligned sets of structurally related proteins. PMID- 8727319 TI - The three-dimensional structure of Escherichia coli porphobilinogen deaminase at 1.76-A resolution. AB - Porphobilinogen deaminase (PBGD) catalyses the polymerization of four molecules of porphobilinogen to form the 1-hydroxymethylbilane, preuroporphyrinogen, a key intermediate in the biosynthesis of tetrapyrroles. The three-dimensional structure of wild-type PBGD from Escherichia coli has been determined by multiple isomorphous replacement and refined to a crystallographic R-factor of 0.188 at 1.76 A resolution. the polypeptide chain of PBGD is folded into three alpha/beta domains. Domains 1 and 2 have a similar overall topology, based on a five stranded, mixed beta-sheet. These two domains, which are linked by two hinge segments but otherwise make few direct interactions, form an extensive active site cleft at their interface. Domain 3, an open-faced, anti-parallel sheet of three strands, interacts approximately equally with the other two domains. The dipyrromethane cofactor is covalently attached to a cysteine side-chain borne on a flexible loop of domain 3. The cofactor serves as a primer for the assembly of the tetrapyrrole product and is held within the active site cleft by hydrogen bonds and salt-bridges that are formed between its acetate and propionate side groups and the polypeptide chain. The structure of a variant of PBGD, in which the methionines have been replaced with selenomethionines, has also been determined. The cofactor, in the native and functional form of the enzyme, adopts a conformation in which the second pyrrole ring (C2) occupies an internal position in the active site cleft. On oxidation, however, this C2 ring of the cofactor adopts a more external position that may correspond approximately to the site of substrate binding and polypyrrole chain elongation. The side-chain of Asp84 hydrogen-bonds the hydrogen atoms of both cofactor pyrrole nitrogens and also potentially the hydrogen atom of the pyrrole nitrogen of the porphobilinogen molecule bound to the proposed substrate binding site. This group has a key catalytic role, possibly in stabilizing the positive charges that develop on the pyrrole nitrogens during the ring-coupling reactions. Possible mechanisms for the processive elongation of the polypyrrole chain involve: accommodation of the elongating chain within the active site cleft, coupled with shifts in the relative positions of domains 1 and 2 to carry the terminal ring into the appropriate position at the catalytic site; or sequential translocation of the elongating polypyrrole chain, attached to the cofactor on domain 3, through the active site cleft by the progressive movement of domain 3 with respect to domains 1 and 2. Other mechanisms are considered although the amino acid sequence comparisons between PBGDs from all species suggest they share the same three dimensional structure and mechanism of activity. PMID- 8727320 TI - Study of global motions in proteins by weighted masses molecular dynamics: adenylate kinase as a test case. AB - The weighted masses molecular dynamics (WMMD) technique is applied to the protein adenylate kinase. A novel set of restraints has been developed to allow the use of this technique with proteins. The WMMD simulation is successful in predicting the flexibility of the two mobile domains of the protein. The end product of the simulation is similar to the known open and AMP bound forms of the enzyme. The biological relevance of the restraints used and potential methods of improving the technique are discussed. PMID- 8727321 TI - On the sensitivity of MD trajectories to changes in water-protein interaction parameters: the potato carboxypeptidase inhibitor in water as a test case for the GROMOS force field. AB - A critical evaluation is presented of the sensitivity of the results of molecular dynamics simulations of proteins to changes in the parameters describing water protein and protein-protein van der Waals interactions in the GROMOS force field. The origin of the van der Waals and electrostatic parameters of the GROMOS standard force field is reviewed, and possible weaknesses are discussed. Four alternate sets of van der Waals parameters for the oxygen types of the GROMOS force field that have been suggested by different authors are then tested against the original force field. Six 500 ps molecular dynamics simulations of the potato carboxypeptidase inhibitor (PCI) in solution using the different parameter sets are analyzed and the results compared with the available X-ray and NMR data. It is shown that the behavior of the molecular system is very sensitive to changes in the van der Waals parameters of the oxygens, especially when affecting the interactions between water and aliphatic or aromatic groups. It is also shown that correction of just the repulsive van der Waals parameter of the water oxygen for its interactions with nonpolar groups is sufficient to correct the main deficiency of the original GROMOS parameter set. Nevertheless, the present study suggest that further refinement of the current parameters is still needed for a proper representation of nonbonded interactions. PMID- 8727322 TI - The rate-limiting step in the folding of the cis-Pro167Thr mutant of TEM-1 beta lactamase is the trans to cis isomerization of a non-proline peptide bond. AB - The stability and kinetics of unfolding and refolding of the P167T mutant of the TEM-1 beta-lactamase have been investigated as a function of guanidine hydrochloride concentration. The activity of the mutant enzyme was not significantly modified, which strongly suggests that the Glu166-Thr167 peptide bond, like the Glu166-Pro167, is cis. The mutation, however, led to a significant decrease in the stability of the native state relative to both the thermodynamically stable intermediate and the fully unfolded state of the protein. In contrast to the two slower phases seen in the refolding of the wild type enzyme, only one phase was detected in the refolding of the mutant, indicating a determining role of proline 167 in the kinetics of folding of the wild-type enzyme. The former phases are replaced by rapid refolding when the enzyme is unfolded for short periods of time, but the latter is independent of the time of unfolding. The monophasic refolding reaction of the mutant is proposed to reflect mainly the trans-->cis isomerization of the Glu166-Thr167 peptide bond. PMID- 8727323 TI - Purification, stabilization, and crystallization of a modular protein: Grb2. AB - We report here the purification and the crystallization of the modular protein Grb2. The protein was expressed as a fusion with glutathione-S-transferase and purified by affinity chromatography on glutathione agarose. It was apparent from reverse phase chromatography that the purified protein was conformationally unstable. Instability was overcome by the addition of 100 mM arginine to the buffers. Because Grb2 appeared to be extremely sensitive to oxidation, crystallization experiments were performed with a dialysis button technique involving daily addition of fresh DTT to the reservoirs. The presence of 8 to 14% glycerol was necessary to obtain monocrystals. These results are discussed in relation with the modular nature of Grb2. PMID- 8727324 TI - Molecular docking using surface complementarity. AB - A method is described to dock a ligand into a binding site in a protein on the basis of the complementarity of the intermolecular atomic contacts. Docking is performed by maximization of a complementarity function that is dependent on atomic contact surface area and the chemical properties of the contacting atoms. The generality and simplicity of the complementarity function ensure that a wide range of chemical structures can be handled. The ligand and the protein are treated as rigid bodies, but displacement of a small number of residues lining the ligand binding site can be taken into account. The method can assist in the design of improved ligands by indicating what changes in complementarity may occur as a result of the substitution of an atom in the ligand. The capabilities of the method are demonstrated by application to 14 protein-ligand complexes of known crystal structure. PMID- 8727325 TI - Accessing the Kabat antibody sequence database by computer. AB - The Kabat antibody sequence database has for many years been the primary site for depositing sequence information on antibodies and other proteins of immunological interest. The chief drawback of this database has been that it has only been available in the form of a printed book (Kabat et al., Sequences of Proteins of Immunological Interest, 1991). These data have recently become available on the global computer Internet, but no method of searching the data has, as yet, been provided. Here, the development of a specialized database program for accessing the antibody data is described. This database software has been made accessible over the World Wide Web, together with a program which allows a novel antibody sequence to be tested against the Kabat sequence database, to identify unusual features of an antibody sequence which may represent cloning artifacts or sequencing errors. PMID- 8727326 TI - Crystallization of a family 8 cellulase from Clostridium thermocellum. AB - The catalytic domain of cellulase CelA, a family 8 glycohydrolase from C. thermocellum, has been crystallized in the orthorhombic space group P2(1)2(1)2(1) with unit cell dimensions a = 50.12 A, b = 63.52 A, c = 104.97 A. The diffraction pattern extends beyond 1.5 A resolution. PMID- 8727327 TI - Crystallization and preliminary crystallographic analysis of RepA1, a replication control protein of the RepFIC replicon of enterotoxin plasmid EntP307. AB - RepA1 protein is essential for replication of the RepFIC replicon of enterotoxin plasmid EntP307 and is thought to interact directly with the origin of replication. We have purified RepA1 from an over-producing expression system and have prepared single crystals using a macroseeding technique. The crystals belong to space group P2(1)2(1)2(1) or P2(1)2(1)2, with cell dimensions a = 61 A, b = 67 A, and c = 243 A. They diffract X-rays to 3.3 A resolution and probably contain two 40,000 molecular weight RepA1 molecules per asymmetric unit. PMID- 8727328 TI - Crystallization and preliminary X-ray analysis of Escherichia coli methionyl tRNA(fMet) formyltransferase. AB - Methionyl-tRNA(fMet) formyltransferase from Escherichia coli, a monomer of 34kDa, was overexpressed from its cloned gene fmt (Guillon, J.M., Mechulam, Y., Schmitter, J.M., Blanquet, S., and Fayat, G., J. Bacteriol. 174:4294-4301, 1992) and crystallized using ammonium sulphate as precipitant. The crystals are trigonal and have unit cell parameters a = b = 151.0 A, c = 81.8 A. They belong to space group P3(2)21 and diffract to 2.0 A resolution. The structure is being solved by multiple isomorphous replacement. PMID- 8727329 TI - Managed care dollars: from patient care to CEOs' paychecks. PMID- 8727330 TI - Tennis calf or tennis leg. PMID- 8727331 TI - Treatment of posterior tibial tendon dysfunction with flexor digitorum longus tendon transfer and calcaneal osteotomy. AB - We treated 32 patients with stage-II posterior tibial tendon dysfunction with calcaneal osteotomy and flexor digitorum longus tendon transfer. These 32 patients (29 women, 3 men) had an average age of 58 years (range, 46 to 73 years) and had been symptomatic for an average of 2.5 years (range, 1 to 8 years) before surgical correction. The indication for surgery was the presence of medial foot pain refractory to nonoperative treatments, including shoewear modifications, orthoses, and bracing. All patients were examined at a mean of 20 months (range, 14 to 48 months) after surgery. Functional and radiographic examinations were performed for each patient and the American Orthopaedic Foot and Ankle Society (AOFAS) foot rating scale was used. Of the 32 patients, 30 were satisfied with the outcome of surgery, had improved function, and exhibited radiographic correction of the foot deformity. The AOFAS score improved from a preoperative mean of 48 points (range, 23 to 76) to a postoperative mean of 84 points (range, 68 to 92). In one patient, treatment failed, necessitating a triple arthrodesis for worsening deformity. The short-term results of this procedure are encouraging. Most patients (94%) experienced pain relief, had improvement in the arch of the foot, and were able to wear regular shoes without orthotic support. In order to correct deformity and provide substantial relief of foot pain and dysfunction, a medial translational calcaneal osteotomy was performed in addition to a flexor digitorum longus tendon transfer for management of stage-II posterior tibial tendon dysfunction. PMID- 8727332 TI - Bone graft in hindfoot arthrodesis: allograft vs autograft. AB - From 1990 to 1992, all arthrodeses of the so-called triple joints, combined or isolated and performed by one surgeon, were reviewed to allow a minimum follow up of 18 months from the time of the index procedure. Type of bone graft was selected based on the patients' decision after the risks of each were explained by the surgeon. There were 37 patients undergoing 41 procedures (4 bilateral) including double and triple arthrodesis, as well as isolated subtalar fusions. Twenty-nine of these were performed for either degenerative or posttraumatic arthritis. Ten others were performed for joint incongruity resulting from posterior tibial tendon insufficiency, and the rest for hemiparesis or residual clubfoot. All patients were placed into a routine postoperative regimen of casting, bracing, weight bearing, and therapy. They were evaluated based on subjective complaints, physical examination, and postoperative radiographs. Any patient suspected of having a nonunion underwent a computed tomography scan for confirmation. Overall, 24 patients received allografts and 17 iliac crest grafts. There were four nonunions, three of which utilized banked bone graft, with the only nonunion in the autograft group occurring in a patient following open calcaneus fracture. Of the four nonunions, three opted for revision surgery and the only one to choose allograft again incurred the only recurrent nonunion. There were two infections, one in each group, resulting in prolonged intravenous antibiotic therapy. PMID- 8727333 TI - True epineural neurolysis in Morton's neuroma: a 5-year follow up. AB - Morton's neuroma is a frequent cause of metatarsalgia. Neuroma resection was the usual recommended surgical treatment. Failure rate of neurectomy can be as high as 14% to 21%, and treatment of recurrences is difficult. The authors have treated Morton's neuroma by neurolysis since 1985. They present their results in a group of 40 patients with 5 years follow up. Thirty-seven of 40 patients had an excellent result after neurolysis and 35 patients had normal toe sensitivity at the date of examination. Thirty-nine patients stated they would undergo the operation again if necessary. PMID- 8727334 TI - Is prophylactic bracing of the ankle cost effective? AB - The ankle joints of 14 healthy volunteers and 16 patients with unstable ankle joints were tested for their functional and proprioceptive capabilities. All of them were active athletes. Three tests were used: the single-leg stance test, the single-leg jumping course test, and the angle-reproduction test. The influence of three stabilization devices--the lace-on brace (Mikros), the stirrup brace (Aircast), and taping--on the proprioceptivity of stable and unstable ankle joints was evaluated. The scores of the single-leg jumping course test without any stabilizing device ("standard" category) ranged between 8.06 and 13.68 (10.65 + 1.29). In the Mikros (9.95 + 0.99) and Aircast (9.99 + 1.14) brace categories, as well as the tape bandage (10.27 + 0.81) category, better scores were achieved. The differences between "standard vs Mikros" and "standard vs Aircast" revealed a significant reduction of the scores with the orthoses (P < .01). The error rate in the single-leg stance test was within the range of 0 to 16 (5.12 + 2.85) for the standard category. It was lower for the Mikros (3.65 + 2.65) and Aircast (4.17 + 2.59) categories. The error rate was highest in the tape bandage group (5.79 + 3.53). There was a significant difference between "standard vs Mikros" and "standard vs Aircast" regarding injured and non-injured ankle joints (P < .01). The angle-reproduction test showed higher values for the standard (2.36 degrees + 0.97) category than the Mikros (1.46 degrees + 0.72), Aircast (1.62 degrees + 0.91), and taping (1.84 degrees + 0.41) categories. In the standard category, the reproduction error was lower when non-injured ankle joints (2.30 degrees + 1.04) were tested than when unstable ankle joints (2.44 degrees + 0.81) were tested, whereas in all other categories the reproduction error was higher in the group of non-injured joints. According to the literature, applying a prophylactic brace can prevent an estimated 30 ankle sprains per 1000 athletic exposures. PMID- 8727335 TI - Valgus tilting of the ankle joint after subtalar (hindfoot) fusion: complication or natural progression of valgus hindfoot deformity? AB - Six patients treated over a 2-year period were noted to have valgus tilting of the talus at the ankle joint on standing anteroposterior ankle films taken after subtalar (hindfoot) fusions. All patients had significant preoperative hindfoot valgus, but no changes on ankle films. All patients underwent "moldable insitu fusions." Severity and duration of preoperative deformity, as well as obesity, appear to be predisposing factors. Most patients were satisfied with their pain relief despite their persistent valgus deformity. Treatment alternatives may include medial displacement calcaneal osteotomy, lateral column lengthening, combination bone and soft tissue procedures, or even tibial talar calcaneal fusion. PMID- 8727336 TI - The miniarthrotomy technique of ankle arthrodesis: a cadaver study of operative vascular compromise and early clinical results. AB - We present a two-part study of the miniarthrotomy ankle fusion technique for minimally deformed joints: a laboratory investigation of ankle vascularity and a clinical review of the early results. In the laboratory portion, five pairs of cadaver legs were injected with radiographic dye, after which arthrodesis was performed via either the open technique (one leg of each pair) or the miniarthrotomy technique (the contralateral leg of each pair). Two legs in the open arthrodesis group had disruption of the peroneal arterial circulation, but no disruption was identified in the miniarthrotomy group. In the clinical portion of the study, we reviewed the results of 32 patients who had undergone ankle fusion via the miniarthrotomy technique. One patient had a nonunion and two patients had delayed union. The average time to union for the 31 patients was 8 weeks (range: 6 to 22). These results compare favorably to the arthroscopically assisted method of fusion and offers another choice for arthrodesis of minimally deformed ankles. We concluded that with minimal disruption, there is less likelihood of arterial injury and devascularization, which may indirectly have a positive effect on the rate of ankle arthrodesis. PMID- 8727337 TI - Chronic laxity of the subtalar joint. AB - Chronic laxity of the subtalar joint derives from the functional significance of the joint with the talo-calcaneal interosseous ligament as the articular "pivot." The laxity of the subtalar joint is usually associated with traumatic or degenerative pathology of the interosseus ligament. The characteristic pain is present along the sinus tarsi and at times also more proximally along the posterior subtalar joint. Radiographic examination is not significant; however, with arthrography, the normal synovial recess of the posterior subtalar joint corresponding to the sinus tarsi is not visualized. With magnetic resonance imaging it has been possible to document structural alterations of ligament. Reconstruction of the interosseus talo-calcaneal ligament has a reasonable expectation of success. The author presents the results of this treatment in 47 operations (44 patients, 3 bilateral) between January 1982 and June 1992. PMID- 8727338 TI - Long-term results of forefoot arthroplasty in patients with rheumatoid arthritis. AB - Disabling forefoot deformity of rheumatoid origin frequently requires surgical intervention. Twenty-three patients (36 feet) who underwent excision of the metatarsal heads in our unit between 1980 and 1987 were assessed clinically and radiologically (n = 12) and by questionnaire (n = 11) at an average 10.5 years (range: 4 to 15) following surgery. Although the procedure was initially successful at the time of review, the result was classified as unsatisfactory because of restriction of walking ability due to pain in the forefoot area in 56% of patients. Recurrence of the deformity--more frequently involving the great toe -had occurred in 72% of patients, and painful callosities were present in 61%. In the patients examined clinically and radiologically, unsatisfactory results were due mainly to mal-alignment of the great toe and extensor tendon tightness. Hindfoot deformity also significantly contributed to pain in the forefoot area. Diminished arthroplasty space, irregular resection cascade, and development of bony spikes were frequently associated with recurrence and callosities. The result of forefoot arthroplasty deteriorates with time. Failure to maintain a plantigrade great toe, intrinsic weakness, and hindfoot deformity were the main factors contributing to an unsatisfactory result. PMID- 8727339 TI - Long-term results of the modified Evans repair for chronic ankle instability. AB - We treated 19 patients for chronic ankle instability with a modified Evans procedure. All patients were evaluated after an average follow up of 128 months with detailed questionnaire, clinical examination, and stress radiographs. Although the subjective results were reported as 8 excellent, 7 good, and 4 fair, residual pain was reported by 11 patients. There was a significantly increased number of osteophytes in the treated ankle joint. Stress radiographs demonstrated significantly improved stability in the operated ankle joint. Range of motion was significantly reduced in hindfoot inversion. The results showed that the modified Evans procedure achieved sufficient joint stability at the expense of inversion range of motion. This reconstruction method apparently did not prevent the development of arthrosis. PMID- 8727340 TI - Helal metatarsal osteotomy for the treatment of metatarsalgia: a critical analysis of results. AB - We reviewed the results of treatment of 114 feet in 96 patients with pressure metatarsalgia treated with a metatarsal osteotomy performed according to the technique of Helal. Clinical examination was performed according to a standardized evaluation questionnaire using the American Orthopaedic Foot and Ankle Society Lesser Metatarsophalangeal-Interphalangeal Scale. The results were correlated with number of metatarsal osteotomies, the length of the follow up, the age of the patient, and type of additional procedures performed. Sixty-one percent of the patients rated the operation as excellent or good. Patient age and number of osteotomies did not influence the results; however, the length of time following surgery and associated Keller-Brandes resection arthroplasty influenced the outcome negatively. There was a high incidence of increased transfer of weight to adjacent metatarsals. We concluded that the Helal type metatarsal osteotomy is unlikely to predictably achieve symptom relief over a long period. PMID- 8727341 TI - Plantar forefoot ulceration with equinus deformity of the ankle in diabetic patients: the effect of tendo-Achilles lengthening and total contact casting. AB - Between 1993 and 1995, 93 neuropathic diabetes mellitus patients with foot ulcers underwent a total contact cast (TCC) protocol. A randomly chosen group of 21 patients (Group I) demonstrated ulcer healing in a mean time of 43.5 days. Despite 9 weeks of TCC, 15 patients (Group II) with forefoot ulcers failed to heal. Physical examination of Group I revealed plantarflexion/dorsiflexion range of motion of the ankle of 33.8 degrees / 1.9 degrees compared to 32.3 degrees / 10.5 degrees of Group II, demonstrating an ankle equinus deformity and limited joint motion. Group II patients underwent a correction of the equinus deformity with percutaneous tendo-Achilles lengthening (TAL), followed by a TCC. All but one ulcer (93.3%) healed within 39.4 days. Four (19.0%) ulcers recurred (at the same site) in Group I, compared to none in Group II at the latest follow up of 17.3 months. Surgical correction with percutaneous TAL and TCC results in healing of forefoot ulcer and helps prevent ulcer recurrence. PMID- 8727342 TI - Long-term follow up of talus fractures. AB - Eighty-six patients who had experienced a fracture of the talus more than 10 years previously were assessed. The fracture had occurred in the neck of the talus in 52 feet and in the body of the talus in 27 feet. The fracture was complicated by dislocation in 47 feet, and aseptic necrosis had developed in 33 feet. The outcome was rated as excellent in 20 feet, good in 43, fair in 18, and poor in 7. The cause of poor outcome in the late stage was posttraumatic osteoarthritis secondary to avascular necrosis and incongruity of the joint surface. Early active exercise without weight bearing is recommended to prevent contracture and bone atrophy due to disuse, which may promote osteoarthrosis. Arthrodesis is recommended in patients with pain because the outcome is greatly improved by arthrodesis, even when performed more than 10 years after the injury. PMID- 8727343 TI - [A study of the significance of serum beta 2-microglobulin levels in patients with multiple myeloma--analyzes as a marker of renal dysfunction and as a marker of tumor cell mass]. AB - Renal involvement known as a myeloma kidney is often observed in patients with multiple myeloma (MM). This complication has been recognized as one of the most important prognostic factors in this disease. Serum beta 2-microglobulin (S beta 2-m) has been also recognized one of the prognostic factor that reflects both a glomerular infiltration rate and a volume of neoplastic cells, because beta 2-m usually can be produced by the neoplastic lymphoid cells. To clarify the significance of the S beta 2-m in MM, we compared S beta 2-m levels and the clinical stage, with another clinical parameters for renal function such as 24 hr creatinine clearance (24 hr Ccr), N-acetyl-beta-glucosaminidase (NAG) levels in urine and serum alpha 1-microglobulin (S alpha 1-m) levels. The elevated S beta 2 m levels were commonly observed not only in patients with stage IIIB, but also in stage IA, IIA and IIIA (76%) who have normal renal function judged by the serum nitrogen or creatine levels. S beta 2-m levels correlated with 24 hr Ccr the most, then correlated with S alpha 1-m levels and less correlated with NAG levels in urine. Although a single elevation of S beta 2-m levels with other normal findings of renal parameters was found only in three out of 30 MM patients, the S beta 2-m levels in these patients did not change after the chemotherapy which had led to the diminution of serum M-protein. Together with these results, it was suggested that the S beta 2-m levels mainly reflect the renal dysfunction even if it stays in the subclinical stage, and reflect less the number of neoplastic cells in MM patients. PMID- 8727344 TI - [Proposals for the objective indices for evaluating the differentiation of leukemic cells by all-trans retinoic acid in the patients with acute promyelocytic leukemia]. AB - All-trans retinoic acid (ATRA) is widely used as a differentiation-inducing agent for acute promyelocytic leukemia (APL). However, one of the problems with this therapy is the difficulty in evaluating the extent of differentiation of leukemic cells. To determine objective indices for differentiation which can be used for judging complete remission, we treated 18 patients with acute promyelocytic leukemia, and performed sequential analysis of bone marrow, karyotype, RAR alpha gene rearrangement and PML-RAR alpha fusion gene transcript. The karyotype, RAR alpha rearrangement and PML-RAR alpha transcript returned to a normal pattern on days 31, 2, 34.8 and 47.6, respectively. We defined and analyzed maturation index (MI) by bone marrow findings as follows; (metamylocyte + band neutrophil + segmented neutrophil) (%)/(myeloblast + promyelocyte + myelocyte) (%). MI increased along with the differentiation of leukemic cells, and the days for MI exceeding 2 well correlated with the normalization of karyotype. These results suggested that MI can be used as an index of differentiation during ATRA therapy. PMID- 8727345 TI - [CD45 gating for flow cytometric analysis of acute leukemia]. AB - Gating of leukemic cells for immunophenotyping by flow cytometry is currently set on two dimensional display of forward scatter (FSC) and side scatter (SSC). However, this gating method can not descriminate leukemic cells from contaminated normal cells (especially normal lymphocytes). Thus, in this paper, we used CD45 monoclonal antibody to detect leukemic cells (CD45dim cells) in conjunction with SSC parameter. This CD45-SSC gating revealed that this method is very useful in AML and ALL, especially leukemia with low percentage blasts. We recommend that this new gating method should be employed for more accurate immunophenotyping of acute leukemia. PMID- 8727346 TI - [Unclassified chronic myeloproliferative disorder (Ph negative/bcr rearrangement negative CML) with late appearing Philadelphia like chromosome]. AB - A 48-year-old male was diagnosed to be unclassified chronic myeloproliferative disorder (UCMPD)/Ph negative bcr rearrangement negative (Ph-/bcr-) CML by hematological, cytogenetical and DNA analyses (Jpn. J. Clin. Hematol. 33(4): 525 531, 1992). Three years and a half after the diagnosis of UCMPD/Ph-bcr- CML, Ph chromosome was observed in 17 of 20 examined cells. Hematological findings showed a transformation into blast crisis. The late appearing of Ph in a case of UCMPD/Ph1-bcr- CML described here is rare. Southern blot analysis using 3' and 5' bcr probe showed no bcr rearrangement. Analyses of BCR/ABL chimeric RNA by RT-PCR method were negative in both of Major- and Minor BCR/ABL chimeric RNA. In the present case it is speculated that Ph is developed as the result of multistep progression and also speculated that the breakpoint at BCR gene differs from Major- and Minor-bcr in usual Ph+CML and de nove Ph+ ALL. Therefore, it may be reasonable that the present case is understood to be a case with late appearing Ph-like chromosome. PMID- 8727347 TI - [Primary myelofibrosis with severe bone pain]. AB - An 80-year-old man was admitted with bone pain, affecting both lower limbs and the dorsa of the feet, in September, 1991. Primary myelofibrosis was diagnosed. Bilateral x-ray films of the tibia, fibula, femur and vertebra revealed osteoporosis, but this was considered an unlikely source of the bone pain. The bone pain subsequently disappeared, but then became severe in both lower limbs and the dorsa of the feet and was accompanied by leukemic transformations in February, 1992. He died of aspiration pneumonia in July, 1992. Autopsy revealed microfractures with osteoporosis and infiltration of leukemic cells in the periosteum of the left tibia, which may have caused the bone pain. This case provides useful information to analyze the cause of bone pain in patients with myelofibrosis. PMID- 8727348 TI - [Hypoplastic mixed lineage leukemia successfully treated with low-dose cytosine arabinoside]. AB - A 54-year-old male was admitted because of dyspnea on exercise. His peripheral blood revealed pancytopenia with severely hypoplastic bone marrow. Bone marrow aspiration showed a marked hypocellular marrow with 62.4% of blast cells. Cytochemical studies showed that peroxidase activity, alpha-nephtyl buthylate esterase activity and PAS reaction were negative, and that only ASD-chroloacetate esterase activity was positive. Surface marker analysis of blast cells showed positive result for CD5, 7, 33 and 34 antigens. The T-cell receptor beta gene was rearranged, but the immunoglobulin H chain gene showed a germ line configuration. Terminal Deoxynucleotydyl Transferase (TdT) was positive, but cellular surface and cytoplasmic immunoglobulin were not recognized. A diagnosis of hypoplastic mixed lineage leukemia was made and treated with low dose cytosine arabinoside, he resulted in complete remission. The relation between hypoplastic leukemia, AML M0 and mixed lineage leukemia was also discussed. PMID- 8727349 TI - [Multiple myeloma associated with phenytoin (diphenylhydantoin) therapy]. AB - The occurrence of multiple myeloma is described in a patient receiving phenytoin (diphenylhydantoin) for 43 years. In this article, five cases of multiple myeloma and one of immunoblastic lymphadenopathy associated with diclonal gammopathy diagnosed after longterm phenytoin therapy are reviewed. These cases are characterized by lymphadenopathy, hepatosplenomegaly, and no or equivocal bone lesions, which are quite unusual in multiple myeloma. Because of the association of lymphomas with phenytoin, the role of the drug in the etiology of multiple myeloma or plasma cell dyscrasias is still in controversy, but highly suspected. It is suggested that a periodic examination of patients receiving phenytoin may be useful in an early detection of M-component. The possibility of reversing multiple myeloma by removal of phenytoin early in the course of the disease should be investigated. PMID- 8727350 TI - [IgD (lambda) multiple myeloma associated with myelofibrosis: an isolated case of nuclear physicist]. AB - A new case of IgD (lambda) multiple myeloma associated with myelofibrosis is described and four from the literature are reviewed. A 66-year-old nuclear physicist who had been diagnosed of having multiple myeloma in 1992 was admitted to the hospital in 1995 because of pancytopenia. A bone marrow biopsy specimen showed nests of myeloma cells with marked fibrotic background. The association of IgD (lambda) multiple myeloma with myelofibrosis was characterized by marked marrow fibrosis and myeloma cell proliferation, without typical features of extramedullary hematopoiesis. Some increase in the bone marrow fibrous tissue is not uncommon in hematologic disorders in which there is a rapid proliferation of marrow cells. What distinguishes these five patients is that their marrow fibrosis was an order of magnitude more extensive. These patients were generally severely anemic and commonly leukopenic and thrombocytopenic. Radiation has been reported as a causal factor in multiple myeloma. In the present case, radiation exposure during his study may have been related to the development of his disease. The correct diagnosis in the case of Dr. Torahiko Terada (1878-1935; physicist, essayist) who died of disseminated bone tumor seems to be multiple myeloma. PMID- 8727352 TI - [Primary myelofibrosis with positive coombs' test responding to prednisolone]. AB - Primary myelofibrosis was diagnosed in a 43-year-old female in 1991, who gradually became transfusion dependent. There was no evidence suggesting connective tissue diseases except for biological false positive STS. Because her direct Coombs' test was positive and serum haptoglobin level was extremely low (< 6mg/dl), intermediate dose (30mg/day) prednisolone therapy was started. Her hemoglobin level and platelet count increased dramatically to a normal level within 3 weeks after the initiation of prednisolone therapy. Bone marrow biopsy performed 6 months later revealed marked recovery of hematopoiesis. Though the effects of corticosteroid therapy in primary myelofibrosis still remain unclear, this therapy might be useful in the treatment of some groups of patients via the correction of immunological aberrations and/or the decrease of bone marrow fiber. PMID- 8727351 TI - [Thrombotic complication in the course of aplastic anemia-paroxysmal nocturnal hemoglobinuria syndrome; possible involvement of dysplasminogenemia (plasminogen Tochigi) in the pathogenesis of thrombosis]. AB - A 44-year-old Japanese man having aplastic anemia (AA)-paroxyamal nocturnal hemoglobinuria (PNH) syndrome was referred to our hospital because of purpuras due to thrombocytopenia in July 1994. He suffered from pneumonia after admission, complicated with cerebral, splenic, and left renal infarction. Pulmonary infaction was also confirmed by perfusion lung scan. He had a plasma plasminogen (PLG) functional activity of 54.4% with a normal level of PLG antigen. The gel isoelectrofocusing pattern of the plasminogen derived from the patient showed 10 normal bands and 10 additional doublet bands with slightly higher isoelectric points than the normal components. Abnormal PLG is converted by urokinase to an inactive two-chain plasmin molecule. These findings were similar to those of a case with dysplasminogenemia (PLG Tochigi) reported by Aoki et al. He was given warfarin for the prevention of thrombosis in December 1994. As of October 1995, these was no recurrence of thrombosis. The cause of thrombosis in the present case have been the association with PNH, predisposition to PLG Tochigi, or the complication of pneumonia. This is the first report of AA/PNH syndrome associated with dysplasminogenemia. PMID- 8727353 TI - [Multiple myeloma presenting as parasellar syndrome and cranial nerve palsies]. AB - Cranial and intracranial locations have been rarely reported in multiple myeloma. Their occurrence as a harbinger of multiple myeloma seems to have a particular significance. In this report, we discuss a case of multiple myeloma presenting as parasellar syndrome and cranial nerve palsies. A 75-year-old woman was admitted to the hospital in June, 1994, with a 3-month history of headache and a 3-week history of diplopia and photophobia. Physical examination revealed right third, fourth and sixth cranial nerve palsies. MRI scan demonstrated a homogeneous, voluminous mass, isointense in T1-weighted images with the cerebral parenchyma and hyperintense in T2-weighted images, occupying the sphenoid sinus and extending within the sella turcica and right cavernous sinus. Lying above the mass and apparently separated from it by a thin rim of hypointensity was a normal pituitary gland. X rays revealed destructive changes of the sella turcica. A minimal disturbance of endocrine function together with a radiologically abnormal pituitary fossa indicated that the primary lesion might lie outside the pituitary fossa. A diagnosis of IgG-kappa type multiple myeloma was made by pertinent laboratory studies. She received local radiation to the intracranial mass (50 Gy) and conventional chemotherapy. Sixteen months after the therapy she is in good health. PMID- 8727354 TI - [Five cases of hereditary high red cell membrane phosphatidylcholine hemolytic anemia in three families]. AB - Five cases of hereditary high red cell membrane phosphatidylcholine hemolytic anemia in three families were described. All cases were clinically manifested by jaundice and splenomegaly. Hemolysis was evident from indirect hyperbilrubinemia, reticulocytosis and decrement of serum haptoglobin. Red blood cells showed morphological abnormalities such as poikylocytosis, anisocytosis and target cells on blood smears. Both direct and indirect Coombs' tests were negative. Ham test, sugar water test and hemoglobin electrophoresis showed no abnormalities. Osmotic fragility test showed decreased membrane fragility. Lipid analysis of red cell membrane showed increment of phosphatidylcholine content and decrement of sphingomyelin content, although plasma lipids were essentially normal. Influx and efflux of sodium through the red cell membrane were both increased. Splenectomy was performed without effect on one patient and the mother of other patients. PMID- 8727355 TI - [Surgical treatment of biliary tract neoplasm]. PMID- 8727356 TI - [Changes in magnetic coil orientation affect the stimulation effects of human peripheral nerve]. AB - Transcranial magnetic stimulation is a non-invasive method for assessing the motor function in humans. It is well established that orientation of the magnetic coil plays an important role on the stimulation effects of the motor cortex. However, there has been a few study regarding the effect of magnetic coil orientation on the efficacy of peripheral nerve stimulation. Therefore, in the present study, I carried out two experiments to clarify the relationship between the magnetic coil orientation and the stimulation effects of human peripheral nerve. First, median nerve was stimulated at the elbow and compound muscle action potentials (CMAPs) of abductor pollicis brevis muscle were recorded in 8 subjects at 4 different directions of the induced current. Two eight-shaped coils, 10 cm and 3.5 cm in outer diameter, were used. Stimulus intensities of big and small coils were 850 V and 90%, respectively. Motor threshold was measured at 4 directions with the big coil by increasing stimulus intensity up to 850 V from 150 V at a step of 50 V. It was found that the amplitude of the CMAP was the greatest in outer direction (toward the radial side) using either the big coil or the small coil. There was no significant difference between the amplitude in outer direction and that of supramaximal electrical stimulation. Motor threshold was the lowest in outer direction. Then, I measured the induced current of the big and small coils in a tank filled with saline that mimicked the forearm. The induced currents by both big and small coils were the largest and the first spatial derivatives of the induced electric field were the greatest in outer direction. These results suggest that the orientation of the eight-shaped coil is important for peripheral nerve magnetic stimulation. The fact that the forearm is a restrictive volume conductor may result in the different effects of coil orientation on the excitement of the peripheral nerve. PMID- 8727357 TI - A clinical study of autogenic training-based behavioral treatment for panic disorder. AB - The present study investigated the effect of autogenic training-based behavioral treatment for panic disorder and identified the predictors of treatment outcome. Thirty-four patients meeting DSM-III-R criteria for panic disorder received autogenic training-based behavioral treatment from October 1981 to December 1994. They were treated individually by the author. The medical records of the patients were investigated for the purpose of this study. The results showed that this autogenic training-based behavioral treatment had successful results. Fifteen patients were cured, nine much improved, five improved, and five unchanged at the end of the treatment. Improvement trends were found as for the severity of panic attack and the severity of agoraphobic avoidance. No consistent findings about predictors emerged when such pretreatment variables as demographics and severity of symptoms were used to predict the outcome. Also, three treatment variables showed useful predictive power. First, practicing the second standard autogenic training exercise satisfactorily predicted better outcomes. Second, application of in vivo exposure was found to be positively associated with the treatment outcome in patients with agoraphobic avoidance. Third, longer treatment periods were associated with better outcomes. These findings suggested that the autogenic training-based behavioral treatment could provide relief to the majority of panic disorder patients. PMID- 8727358 TI - [Computer-assisted image analysis of calcium ion concentration]. PMID- 8727359 TI - Peritubular myoid cells in the testis: their structure and function. AB - Peritubular myoid cells, surrounding the seminiferous tubules in the testis, have been found in all mammalian species, but their organization in the peritubular interstitial tissue varies by species. In laboratory rodents, including rats, hamsters and mice, only one layer of myoid cells is seen in the testis. The cells in these animals are joined by junctional complexes as are epithelial cells. On the other hand, several cellular layers exist in the lamina propria of the seminiferous tubule in the human and some other animals. Myoid cells contain abundant actin filaments which are distributed in the cells in a species-specific manner. In the rat, the filaments within one myoid cell run both longitudinally and circularly to the long axis of the seminiferous tubule, exhibiting a lattice work pattern. The arrangement of the actin filaments in the cells changes during postnatal development, and the disruption of spermatogenesis, such as cryptorchidism, seems to affect further the arrangement of the filaments. Other cytoskeletal proteins, including myosin, desmin/vimentin and alpha-actinin, are also found in the cells. Myoid cells have been shown to be contractile, involved in the transport of spermatozoa and testicular fluid in the tubule. Several substances (prostaglandins, oxytocin, TGF beta, NO/cGMP) have been suggested to affect the contraction of the cell, though the mechanisms of the contraction are still unknown. Recent in vitro studies have demonstrated that the cells secrete a number of substances including extracellular matrix components (fibronectin, type I and IV collagens, proteoglycans) and growth factors (PModS, TGF beta, IGF-I, activin-A). Some of these substances are known to affect the Sertoli cell function. Furthermore, it has been reported that myoid cells contain androgen receptors and are involved in retinol processing. Considering all this, it is evident that peritubular myoid cells not only provide structural integrity to the tubule but also take part in the regulation of spermatogenesis and the testicular function. Their precise roles, however, remain to be solved. PMID- 8727360 TI - Regeneration of fin rays in teleosts: a histochemical, radioautographic, and ultrastructural study. AB - Regeneration followed partial amputation of the tail fin in three representative species of teleosts. A systematic study, using histochemical methods, radioautography and transmission electron microscopy, disclosed the essentials of the natural history of regeneration of the fin soft-rays. At about 24 hours, epidermal cells had completely covered the cut edge. By the second day an apical epidermal cap was established. Beneath this cap a blastema was formed by about three days after amputation. A collagenous lepidotrichial matrix had begun to develop by 4 days after excision. Actinotrichia were first detected in the regenerates at 5 days. During these processes, the basal lamina of the epidermis played an important morphogenetic role. The cells responsible for the regenerate underwent regressive changes after form and function had been restored. PMID- 8727361 TI - Nitrinergic nerves controlling pacemaker activities of the inner sublayer (P layer) in the canine proximal colon circular muscles. AB - The inner sublayer (P-layer) of the circular muscle coat in the canine proximal colon has been known to produce spontaneous mechanical contractions associated with characteristic electrical activities called slow waves. We recorded the mechanical activities of tissue preparations from this P-layer. Normal Krebs solution (K+; 6 mM) was used as the perfusate. Elevation of extracellular K+ concentrations in the range of 12 mM and 36 mM induced intensified phasic contractions. Administration of an NO-synthase inhibitor, N omega-nitro-arginine methyl ester (L-NAME, 50 microM), enhanced both the spontaneous mechanical rhythms and high extracellular K(+)-induced contractions. Administration of the substrate for NO synthases, L-arginine (400 microM) remarkably suppressed the effects of L-NAME on the amplitude of the spontaneous rhythms and on responses to extracellular high K+. Histological structures of nerves in the P-layer were investigated by an NADPH (nicotinamide adenine dinucleotide phosphate)-diaphorase technique and by the immunohistochemistry of NO-synthases, since NO-producing (nitrinergic) nerves usually, if not always, show a histochemical NADPH diaphorase positive reaction in formaldehyde-fixed specimens, and since features of ganglia and nerve strands in the outer subdivision of the submucosal plexus (plexus submucosus externus; or so-called Henle's plexus) together with the delicate network of nerve terminal varicosities within the P-layer were clearly visualized by this method. The topographical arrangement of nitrinergic nerves supported the view that they produce nitric oxide (NO), being one of the major chemical mediators of the neural control of the spontaneous rhythms in the P layer. PMID- 8727362 TI - The innervation of taste buds in the soft palate of the rat as revealed by methylene blue staining. AB - Light microscopic data on the innervation of taste buds in the soft palate of the rat obtained by methylene blue (MB) supravital staining are presented here for the first time. The taste pore including subpopulations of taste bud cells and nerve fibers were stained. Intragemmal nerve fibers were detected ascending up to the taste pore; this suggests the possibility of additional paracellular pathways in taste transduction directly via nerve fibers. Moreover, relatively large bulbous or laminar enlargements of the intragemmal nerve fibers were found. The branches of these nerve fibers, showing minute spiny protuberances, appeared to anastomose. Some perigemmal nerve fibers were also seen in close apposition to the taste pore. The basal plexus in the vicinity of the taste buds showed enlargements which resembled the Merkel's discs, but were significantly smaller. These structures did not occur in the subgemmal part of the basal plexus. Regarding the oxygen-dependency of the staining reaction, it was speculated that both MB and oxygen were attracted by iron-containing enzymes. Therefore, the strong affinities for MB shown by subpopulations of nerve fibers and taste bud cells were explained by their high requirements for oxygen. Furthermore due to its selectivity for intraepithelial nerve fibers and sensory cells, the method applied here is well-suited for routine use in light microscopy. PMID- 8727363 TI - Evidence of synaptic contacts of nociceptive primary afferent central terminals on GABAergic interneurons in the substantia gelatinosa. AB - Recently, we showed that capsaicin induced the degeneration of not only glomerular CI terminals but also of non-glomerular CI terminals making presynaptic contact with interneuronal soma. Studies of the nature of interneurons making contact by nonglomerular CI terminals should provide important information to facilitate our understanding of the processing of nociceptive impulses in the substantia gelatinosa. The most likely candidate molecule involved in this process in these interneurons is gamma-aminobutylic acid (GABA). Therefore, ultrastructural relationships between nonglomerular CI terminals land GABAergic interneuronal soma in the mouse substatia gelatinosa were examined by an immunocytochemical method using an antibody to GABA. Terminals with the same profiles as the CI terminals, i.e., slender, sinuous and scalloped terminals filled with clear synaptic vesicles, were found to make synaptic contacts with GABA-immunoreactive somata. Thus, nociceptive primary afferents are suggested to modulate pain transmission by themselves via GABAergic neurons in the substantia gelatinosa. PMID- 8727364 TI - Three-dimensional organization of lymphatics in the dog stomach: a scanning electron microscopic study of corrosion casts. AB - The three-dimensional organization of the lymphatics in the dog stomach was studied by scanning electron microscopy of corrosion casts, which were made by direct parenchymal injection of low-viscosity Mercox-resin into the mucosa and the muscular layers. Although the organization of lymphatics in the stomach has been studied by a variety of methods, the origin of the lymphatics and their connection in each layer has not been clearly defined. In this study, using dog stomach because of its structural similarity to the human stomach, we defined the lymphatic structure in all gastric layers, with lymphatics absent in the upper two-thirds of the lamina propria mucosae. They were first encountered at the deepest level of the lamina propria, immediately above the lamina muscularis mucosae. These lymphatics were composed of single-layered irregular meshes. Slender lymphatics arising from this network passed through the lamina muscular is mucosae and drained into the lymphatic plexus, which was composed of thicker lymphatics at the uppermost layer of the submucosa. Lymphatic valves were frequently seen in this plexus. From this plexus, slender connecting lymphatics with valves extended straight downward without lateral communications and drained into the lymphatic plexus at the deepest layer of the submucosa. This latter plexus, composed of large-caliber lymphatics, issued flattened lymphatics which formed a three-dimensional network in the muscular layer. The subserosal lymphatics were composed of thick lymphatics with tortuous courses and drained into the efferent lymphatics. PMID- 8727365 TI - Mesangial cells of the lamprey, Lampetra japonica, store vitamin A. AB - Structure and vitamin A-storage of the mesangial cells in the lamprey kidney are documented using the gold chloride method, fluorescence histochemistry and electron microscopy. Abundant lipid droplets in the cytoplasm of the mesangial cells release intense, but quickly fading vitamin A-fluorescence under a fluorescence microscope and react with gold chloride. Characteristic tubular invaginations of the plasma membrane which usually occur in the lamprey smooth muscle cells are observed in these cells. We discuss similarities of the lamprey mesangial cells with hepatic stellate cells in the lamprey and mammals, such as vitamin A-storage, blood flow-regulation and proliferative potential. To the best of our knowledge, this is the first report on and proof of vitamin A-storage in mesangial cells, providing evidence for a vitamin A-storing cell lineage in vertebrates. PMID- 8727366 TI - Dark neurons in the mouse brain: an investigation into the possible significance of their variable appearance within a day and their relation to negatively charged cell coats. AB - This study aims to investigate the occurrence and nature of dark neurons in the central nervous system under physiological conditions. Mouse brain tissues were perfusion-fixed with paraformaldehyde or glutaraldehyde at 4 h intervals during one day (3:00, 7:00, 11:00, 15:00, 19:00, 23:00). Paraffin sections were stained with the cationic colloidal iron method, and counterstained with nuclear fast red or carbolthionin. The dark neurons were readily distinguishable as their shrunken cell bodies stained densely with nuclear fast red or thionin. Some of the dark cells were coated with perineuronal sulfated proteoglycans; this coat, which formed a smoothly extended meshwork in light cells, presented spicule-like forms in the dark cells. The occurrence of dark cells in the retrosplenial cortex varied by the time of day: the incidence of the dark neurons was low (10-15%) at 11:00, 15:00 and 23:00, while it was significantly high (50-60%) at 3:00 and 19:00. Previous authors have ascribed the occurrence of dark neurons either to artifacts due to inappropriate fixation or to pathological damage. However, the present study strongly suggests that this type of neuron occurs under physiological conditions as reversible changes, and vary over a day, showing distinct peaks. These peaks occurred coincidentally while the mice were awake. Such morphological changes may be involved in the neuronal activation and exhaustion. Our view is consistent with the hypothesis (Tewari and Bourne, 1963) that the neurons take such dark profiles at certain stages of neurosecretion. PMID- 8727367 TI - Immune cell migration through the arterial wall in the murine lung during a pulmonary inflammatory response. AB - Perivascular accumulation of leukocytes in the lung was induced by intratracheal administration of sheep erythrocyte antigen to primed mice. The route of migration of intravascular leukocytes to the perivascular space in the lung, in particular from arteries, and the structure of lymphatic vessels among the aggregated leukocytes were examined by transmission electron microscopy. Leukocytes--lymphocytes, granulocytes, monocytes and macrophages--were demonstrated to adhere to the endothelial surface and to migrate between endothelial cells to reach the internal elastic lamina of arteries. Becoming conspicuously constricted, the leukocytes penetrate through this elastic lamina. They further migrate through the smooth muscle layer to the interstitium, passing through the external elastic laminar region. At 2 days after antigen administration, dilated lymphatic vessels containing large numbers of leukocytes in the lumen and bearing endothelial gaps open to the interstitium began to be seen. The lymphatic walls were more convoluted and richer in pinocytotic vesicles than those prior to antigen challenge. This study confirms the light microscopic findings by Curtis et al. (1990) that arteries, besides veins, venules and capillaries, may represent a major route of inflammatory cell entry into the lung parenchyme in an acute and vigorous immune response. In addition, lymphatic vessels were suggested to be newly formed for the transport of fluid and immune cells from the sites of inflammation in the lung. PMID- 8727368 TI - [DNA diagnosis in myotonic dystrophy]. AB - Myotonic dystrophy (DM) is an autosomal-dominant, multisystemic disorder characterized by myotonia, progressive muscle atrophy and weakness, cardiac conduction defect, mental retardation, and cataracts. The phenotypic expression of DM varies from asymptomatic adults to severely affected neonates with congenital DM (CDM). DM shows genetic anticipation, an increase in disease severity and earlier age of onset in successive generations. The molecular basis of DM mutation is an unstable trinucleotide (CTG) repeat located in the 3' end of a transcript that encodes a myotonin-protein kinase. Normal populations have 5 to about 30 CTG repeats, where DM patients have 50-2,000 such repeats. The CTG repeat number is expanded in DM patients when transmitted from parent to child severity. An approximate correlation has been demonstrated between the degree of CTG repeat expansion and clinical severity. The largest repeat sizes are disclosed in CDM. Furthermore, we presented haplotype analysis of CDM families and disclosed localization of myotonic dystrophy protein kinase in DM muscle. DM kinase mRNA was decreased in various tissues of CDM patient. PMID- 8727369 TI - [Genetic diagnosis of colorectal cancer]. AB - Accumulating evidences that carcinogenesis requires multiple gene alterations of oncogenes and tumor suppressor genes have recently emerged. In addition, genes related to invasion and metastasis are also important in understanding development of colorectal cancer. In this study, clinical significance and application of tumor suppressor genes and invasion related genes such as APC (adenomatous polyposis coli), DCC (deleted in colorectal carcinoma) tumor suppressor genes and invasion related gene, matrilysin were studied. In the mouse tumor induced by mutagen contained in cooked food, PhIP (2-amino-1-methyl-6- phenylimidazo [4,5-b] pyridine), nonsense mutations of APC gene that is similar to human colorectal cancer have been observed. These results suggested the quite interesting issue of mutagen contained in daily food having etiological role of colorectal cancer. DCC gene alteration, decreased expression of DCC mRNA was detected in 60% of advanced colorectal cancer. In all cases with liver metastasis, DCC expression was absent or markedly decreased, a finding that detection of DCC expression have an clinical importance that predicts metastatic potential of colorectal cancer. Matrilysin, the member of MMPs (matrix metalloproteinases) which degrade matrix components such as type IV collagen, laminin or fibronectin. In most of colorectal cancer, matrilysin was overexpressed in tumor cells. Matrilysin-transfected colorectal cancer cells showed more invasive ability in vitro and gained metastatic potential in SCID mice. Suppression of matrilysin expression by treated with all-trans retinoic acid (ATRA) or introduction of anti-sense matrilysin decreased the invasive ability in vitro. This result suggests that matrilysin plays an important role in invasion and metastasis and have a possibility of new anti-invasion therapy. PMID- 8727370 TI - [Gene therapy for cancer]. AB - The strategies of gene therapy for cancer can be classified as: 1. regulation of oncogenes and anti-oncogenes, 2. immunogenetherapy, 3. support of chemotherapy, 4. suicide gene therapy, and 5. gene marking. The first one is the strategy to inhibit the expression of oncogenes by their antisenses or rhybozymes, or to introduce anti-oncogenes into those tumor cells with the inactivate effector cells (lymphocytes) by transducing cytokine genes, etc., followed by retransfering the gene-modified effector cells to patients (adoptive immunotherapy). The other one is to augment antigenicity of tumor cells. The immunogenetherapy method has been widely used for 70% of gene therapy of human cancer, because cells can be transduced ex vivo. The anti-tumor effects of human gene therapy using a GM-CSF gene by Muligan et al. or an IL-2 gene by Tahara and Lotze are expected. The third is the strategy to protect bone marrows from large dose of anti-cancer drug by transducing a multidrug resistance gene into those bone marrow cells or periphen blood stem cells, overcoming dose limiting of the drug. The fourth strategy is to transduce the herpes simplex virus thymidine kinase (KSV-TK) gene for activating the cytocydal prodrug (ganciclovir: GCV) into tumor cells in order to kill the tumor cells themselves following administration of GCV. At present, vectors most widely used for gene transduction are retroviruses and adenoviruses. However, the transduction using these vectors are primarily conducted ex vivo. The direct in vivo gene delivery method to target tumor cells are required. PMID- 8727371 TI - [Safer trials of vaccine and gene therapy against HIV]. AB - HIV, the etiological agent of AIDS, induces depletion of CD4+T cells. Upon primary infection, there is an initial viremia that is followed rapidly by a cell mediated immune response and apparent viral clearance. Thereafter, the persistence of low levels of HIV in the blood for years before the onset of the disease is facilitated by the ability of the virus to establish persistent and latent infection. Notwithstanding the latent form in most population, the clinical stage of disease is significantly associated with all measures of virus load levels. This may be mainly due to the function of HIV to induce apoptosis in a patients' uninfected CD4+T cells, as a bystander effect. Thus, the mechanisms that latently infect in the cells after HIV infection and stimulate active replication of HIV from the latency are essential for an understanding of the pathogenicity of the disease. Therefore, prophylactic and therapeutic trials should be focused to their effect to reduce the HIV load level. PMID- 8727372 TI - [Gene therapy for adenosine deaminase deficiency]. AB - Severe combined immunodeficiency (SCID) due to adenosine deaminase (ADA) deficiency is a fatal recessive disorder caused by mutations in the gene encoding ADA. Based on the first clinical trial of two young girls with ADA-deficient SCID by recombinant retrovirus-mediated gene transfer at the National Institute of Health of USA, we prepared to treat a four-year-old boy with ADA-deficient SCID who had been treated with PEG-ADA for 3 years. Approval to perform the clinical trial of gene therapy by using his peripheral blood T lymphocytes as the target and recombinant retroviral vector (LASN) as the vector for ADA gene transfer was obtained from both of the Ministry of Health and Welfare and the Ministry of Education, Science, Sports and Culture on 13 February, 1995. The first clinical trial of gene therapy for the patient was initiated on 1 August 1995. He received 8 x 10(8) LASN-transduced lymphocytes in an injection administered intravenously on 8 August and 2.5 x 10(9) transduced lymphocytes on 4 September without any side reactions. The procedure, safety and efficacy of clinical trial of gene therapy were discussed. PMID- 8727373 TI - [Analysis of a novel gene expressed in apoptotic T-cell hybridoma treated with dexamethasone]. AB - Developmentally and physiologically controlled cell deaths are called programmed cell death (PCD), and distinguished from accidental cell death while apoptosis is defined morphologically as a certain type of cell death (i.e. chromatin condensation, DNA fragmentation, organella compaction, and blebing of cell and nuclear membranes). Apoptosis is often associated with PCD. Most of cells undergoing PCD have been shown to require de novo synthesis of mRNA and protein. A number of genes have been reported to be expressed in apoptotic cells. In the immune system programmed cell death plays an important role in negative selection of immature T cells. Situations that result in programmed cell death include the withdrawal of cytokine from cultures of cytokine dependent cells, lysis of target cells by cytotoxic T cells, and activation of T cells. In T cells, another well described means of inducing apoptosis is an exposure to glucocorticoids. We cloned a novel gene expressed in apoptotic 2B4. 11 mouse T-cell hybridoma treated with steroid. This gene was not detected in vivo, but was so in mouse thymocytes and splenocytes cultured with dexamethasone and IgM stimulated mouse preB cell line WEHI-231. Thirty-six amino acids, position 2-37, has a 37% homology to mouse intracisternal A-particle gag p27 protein. PMID- 8727374 TI - [Studies on nuclear DNA content in bladder cancers using flow cytometry]. AB - This study was designed to evaluate the clinical relevance between the DNA ploidy and histopathology, and the incidence of the DNA heterogeneity in patients with bladder cancers. Flow cytometry(FCM) was used to study the DNA ploidy in 63 patients who underwent total cystectomy. The DNA ploidy and DNA index were analyzed by FCM in total 478 paraffin embedded samples (7.6 samples per case on the average). The DNA ploidy of 52 bladder cancers, that had coexisted after total cystectomy, showed that 24 cases, 46% were DNA aneuploid and 18 cases, 35% had DNA heterogeneity. The other 11 cases which were found to be pT0 after cystectomy were all DNA diploid. There were significantly good correlation among DNA ploidy pattern and intravesical involvement (lymph-duct involvement and venous involvement), but were not among the DNA ploidy pattern and tumor grade and stage. With regard to the evaluation of two vertically devided samples of tumors, DNA aneuploid had been not always recognized in the deeper samples, therefore, the author did not determine that there was good correlation between the DNA ploidy and the tumor invasion. In lymph node metastases, the 39 diploid tumors had given rise to lymph node metastases in only 5 cases(13%), whereas 11 cases(46%) of the 24 aneuploid tumors had metastasized(p < 0.01). Eleven of 16 lymph node metastases were DNA diploid and the others were DNA aneuploid. These data suggest that although the incidence of DNA heterogeneity in bladder cancers (35%) is thought to be relatively small, the DNA ploidy will be able to be the important prognosticating factor in bladder cancers. PMID- 8727375 TI - [Influence of total parenteral nutrition in immature rats: effect of glutamine infusion on bowel integrity]. AB - Total parenteral nutrition (TPN) is accepted important therapeutic adjunct in spite of many complications for management of pediatric patients who aren't allowed to eat. Recently, bacterial translocation was added to the complications of TPN. The purpose of this study was to examine the influence of TPN on the gut in immature rats. 65g rats were randomized to one of four groups: Group C (control) received food and water ad libitum. Group P (TPN) received standard TPN solution. Group G (TPN+GLT) received glutamine (7g/400ml of TPN). Group E (TPN with enhanced protein) received TPN solution with enhanced protein (18.5g/400ml), without GLT. CFU of E. coli in mesenteric lymphnodes was significantly higher in group P than in other 3 groups at 5 days. Hepatic glutathione was significantly higher in group G than in group P and group E at 7 days. Weight of wet intestine was the highest in group C in all groups, and significantly higher in group G than in group P and group E at 3 and 7 days. Mucosal protein of group C was the highest of those of 4 groups. That of group G was significantly higher than those of group P and G in 5 and 7 days. Mucosal thickness and villous height were the highest in group C in 4 groups. Mucosal thickness was significantly higher in group G than in group P and group E at 5 and 7 days. Villous height was significantly higher in group G P and group E at all days. These results suggest that TPN promotes intestinal atrophy from early days after TPN in immature rats, that glutamine might play a role in maintenance of structural integrity of intestine, and that glutamine would prevent the bacterial translocation. PMID- 8727376 TI - [Anti-tumor effects of an angiogenesis inhibitor, TNP-470 (AGM-1470) alone and in combination with cisplatinum, and changes in serum copper levels in liver cancer bearing rats]. AB - Effects of a newly developed angiogenesis inhibitor, TNP-470 (AGM-1470) alone and in combination with cisplatinum on tumor-bearing rats were investigated. Wistar King A male rats were subcutaneously inoculated with 10(7) cultured syngeneic liver cancer cells that had been induced by oral intake of 3'-methyl-4 dimethylaminoazobenzen, and were used as tumor-bearing rats. Tumor sizes and changes in serum copper levels were measured after administration of TNP-470. Administration of TNP-470 (5, 10, 20 or 30mg/kg, s.c., daily for 7 days) inhibited tumor growth in a dose-dependent manner. The suppressive effects were the same in the rats which were administered TNP-470 (20mg/kg, s.c., daily for 7 days) and cisplatinum (0.5 mg/kg, s.c., daily for 7 days) simultaneously, and rats administered cisplatinum (0.5mg/kg, s.c., daily for 7 days) following TNP 470 (20mg/kg, s.c., daily for 7 days). These inhibitory effects were almost the same as that of cisplatinum (0.5mg/kg, s.c., daily for 7 days) alone. While, administration of TNP-470 (20mg/kg, s.c., daily for 7 days) following cisplatinum treatment (0.5 mg/kg, s.c., daily) showed markedly higher anti-tumor effects, compared with these groups. Administration of TNP-470 caused elevation of serum copper levels in normal rats as well as tumor-bearing rats with the same degree. Serum copper levels remained normal after discontinuation of TNP-470 in normal rats, while in tumor-bearing rats, it decreased during the first week and re elevated in 2 to 3 weeks after discontinuation of TNP-470. This re-elevation of serum copper levels was related to rapid tumor growth after discontinuation of TNP-470. Furthermore, there was also a positive correlation between serum copper levels and capillary density in the tumor. In conclusion, TNP-470 had anti-tumor effect in a dose-dependent manner against a rat liver cancer, with markedly higher effects when it was administered following cisplatinum. Serum copper levels after discontinuation of TNP-470 treatment might indicate re-proliferation of the capillaries in the tumor tissue. PMID- 8727377 TI - [Analysis of the iron-related proteins during proliferation and differentiational change of human hepatoblastoma cells (HepG2)]. AB - The modulation of cellular iron metabolism depends on the expression of ferritin and transferrin receptor (TfR). These proteins are regulated post transcriptionally by interaction of specific sequences, termed iron responsive elements (IRE) which are located in the 5' untranslated region (UTR) of ferritin mRNA and 3'UTR of TfR mRNA and a specific cytosolic protein, termed iron responsive element-binding protein (IRE-BP). The liver plays an important role in iron metabolism as a major organ of iron storage. In hepatocytes iron is taken up in the form of transferrin and intercellular iron levels are regulated by IRE/IRE BP interaction. In spite of the important role of IRE/IRE-BP interaction, the role of it for proliferation and differentiational change in hepatoma cells is not well known. In this study, the author has investigated IRE/IRE-BP interaction and its target protein expression during proliferation and differentiational change of human hepatoblastoma cells (HepG2). This study revealed that the binding activity of IRE-BP to IRE was increased when HepG2 cells were cultured with an iron-chelating agent, deferoxamine (4.5 microM), and it was decreased when cultured with transferrin (100 micrograms/ml) during both proliferation and differentiational change induced by sodium butyrate. Also, the change of TfR expression was correlated with that of IRE-BP. These findings suggest that IRE/IRE-BP interaction may play an important role in the expression of TfR and cellular iron metabolism during proliferation and differentiational change of HepG2 cells. PMID- 8727378 TI - [Influence of aging on the unilateral and the contralateral amygdaloid kindling in rats]. AB - To investigate the seizure susceptibility in the rat after the mature period, the unilateral (primary site, left) and the subsequent contralateral (secondary site, right) amygdaloid kindling were examined in 2-3-month-old (n = 12, group A), 12 month-old (n = 11, group B), and 18-month-old (n = 7, group C) male Sprague Dawley rats. At both of the primary and the secondary sites, the number of stimulations required to produce a generalized convulsion with falling (stage 5) increased as the age progressed, and there was a significant difference in the number of stimulations to reach stage 5 between groups A and B, and between groups A and C. These findings suggest that the age-related retardation of the development of amygdaloid kindling reflects aging changes of synaptic transmission. On the other hand, positive transfer effect (a facilitory effect of the primary site amygdaloid kindling on the development of the secondary site amygdaloid kindling) was observed in all of the three groups. This results suggests that both in and after the mature period, the secondary site amygdaloid kindling in the rats utilizes proconvulsant neuroplastic changes in the brainstem that has been already established by the primary site amygdaloid kindling. PMID- 8727379 TI - [Sequential observations of brain edema with proton magnetic resonance imaging and spectroscopy]. AB - The purpose of this study was to assess the relationship between morphological and metabolic changes in brain edema using proton magnetic resonance systems. The serial changes during the first 24 hours in the cold-injury trauma rat brain model were investigated by proton magnetic resonance imaging (1H MRI) and high resolution proton MR spectroscopy (1H MRS). We also analyzed the efficacy of AVS 1,2-bis (nicotinamide)-propane which can scavenge free radicals to the edema in this experiment. The edema was developing extensively via the corpus callosum in ipsi- and contralateral hemispheres as shown by gradually increased signal intensity on 1H MRI. 1H MRS initially showed accumulation of acetate and lactate, and transient increasing of glutamine. After 24 hours, the increased glutamine decreased below the control, alanine increased, and N-acetyl asparatate decreased with the edema development. AVS-treatment significantly suppressed edema development, increases of lactate and alanine and decreases of N-acetyl asparatate. We suggest that the cold-induced lesion contains anaerobic glycolysis deterioration and results in severe brain tissue breakdown. AVS is proved valuable for the treatment of this edema lesion. Clinical 1H MRS showed prolonged lactate elevation and significant decreases of other metabolites in human ischemic stroke edema. In peritumoral edema, decreased N-acetyl asparatate gradually improved, and slightly elevated lactate disappeared after tumor removal. 1H MRS feasibly characterizes the ischemic and peritumoral edema and makes a quantitative analysis in human brain metabolism. We believe the combined 1H MRI and MRS study is a practical method to monitor the brain conditions and will make it easy and possible to find new therapeutic agents to some brain disorders. PMID- 8727380 TI - Activity patterns of the B31/B32 pattern initiators innervating the I2 muscle of the buccal mass during normal feeding movements in Aplysia californica. AB - 1. B31 and B32 are pattern-initiator neurons in the buccal ganglia of Aplysia. Along with the B61/B62 neurons, B31/B32 are also motor neurons that innervate the 12 buccal muscle via the I2 nerve. This research was aimed at determining the physiological functions of the B31/B32 and B61/B62 neurons, and of the I2 muscle. 2. Stimulating the I2 muscle in the radula rest position produces radula protraction. In addition, in behaving animals lesioning either the muscle or the I2 nerve greatly reduces radula protraction. 3. During buccal motor programs in reduced preparations, B31/B32 and B61/62 fire preceding activity in neuron B4, whose firing indicates the onset of radula retraction. In addition, during both ingestion-like and rejection-like patterns the activity in the I2 nerve is correlated with protraction. 4. B31/B32 fire at frequencies of 15-25 Hz. Neither B31/B32 nor B61/B62 elicit facilitating end-junction potentials (EJPs) and electromyograms (EMGs) in the I2 muscle. EMGs from B31/B32 are smaller than those from B61/B62. B31/B32 and B61/B62 innervate all areas of the muscle approximately uniformly. 5. In behaving animals, EMGs consistent with B31/B32 activity are seen in the I2 muscle during the protraction phase of biting, swallowing, and rejection movements. In addition, the I2 muscle receives inputs that cannot be attributed to either the B31/B32 or B61/B62 neurons, either because the potentials are too large, firing frequencies are too low, or a prominent facilitation is seen. Such potentials are associated with lip movements, and also with radula retraction. 6. EMGs were recorded from the I2 muscle during feeding behavior after a lesion of the I2 nerve. Animals that had severe deficits in protraction showed no activity consistent with B31/B32 or B61/B62, but did show activity during retraction. 7. Our data indicate that the I2 muscle and the B31/B32 motor neurons are essential constituents contributing to protraction movements. Activity in these neurons is associated with radula protraction, which occurs as a component of a number of different feeding movements. The I2 muscle may also contribute to retraction, via activation by other motor neurons. PMID- 8727381 TI - B64, a newly identified central pattern generator element producing a phase switch from protraction to retraction in buccal motor programs of Aplysia californica. AB - 1. Buccal motor programs in Aplysia are characterized by two phases of activity, which represent protraction and retraction of the radula in intact animals. The shift from protraction to retraction is caused by synaptic activity inhibiting neurons that are active during protraction and exciting neurons that are active during retraction. 2. B64, a newly identified neuron present bilaterally in the buccal ganglia, is partially responsible for the phase shift. Stimulating a single B64 causes bilateral inhibition of neurons B31/B32 and other neurons active during protraction and cause bilateral excitation of neurons B4/B5 and other neurons active during retraction. B64 is active throughout retraction. The amplitude and waveforms of the synaptic potentials caused by firing B64 are similar, but not identical, to those seen during retraction. 3. Some of the effects of B64 on B31/B32 and on B4/B5 are monosynaptic, as shown by their maintained presence in high divalent cation seawater, which blocks polysynaptic activity. 4. A brief depolarization of B64 leads to a long-lasting depolarization and firing. The ability of B64 to respond in this way is at least partially caused by an endogenous plateau potential, as this property is still seen after synaptic transmission is blocked. 5. Hyperpolarization of B64 bilaterally and preventing the somata from firing unmasks a large excitatory postsynaptic potential in B64. This procedure does not block the shift from protraction to retraction, indicating that spiking in the B64 somata is not necessary for the phase shift. 6. The firing pattern and synaptic connections of B64 are consistent with the hypothesis that the neuron is part of a central pattern generator underlying buccal motor programs. B64 is monosynaptically inhibited by neurons that are active along with B31/B32, which are responsible for producing the protraction phase of a buccal motor program. During the later portion of the protraction phase B64 is excited. In addition, firing B64 can phase advance and phase delay buccal motor programs. 7. Regulating the firing of B64 can regulate the expression of buccal motor programs. Stimulation of B64 at frequencies of 0.5 1.0 Hz leads to complete inhibition of buccal motor programs, whereas steady state depolarization of B64 can lead to repetitive bursts of activity. PMID- 8727382 TI - Information theoretic analysis of dynamical encoding by four identified primary sensory interneurons in the cricket cercal system. AB - 1. The stimulus/response properties of four identified primary sensory interneurons in the cricket cercal sensory system were studied using electrophysiological techniques. These four cells are thought to represent a functionally discrete subunit of the cercal system: they are the only cells that encode information about stimulus direction to higher centers for low intensity stimuli. Previous studies characterized the quantity of information encoded by these cells about the direction of air currents in the horizontal plane. In the experiments reported here, we characterized the quantity and quality of information encoded in the cells' elicited responses about the dynamics of air current waveforms presented at their optimal stimulus directions. The total sample set included 22 cells. 2. This characterization was achieved by determining the cells' frequency sensitivities and encoding accuracy using the methods of stochastic systems analysis and information theory. The specific approach used for the analysis was the "stimulus reconstruction" technique in which a functional expansion was derived to transform the observed spike train responses into the optimal estimate (i.e., "reconstruction") of the actual stimulus. A novel derivation of the crucial equations is presented. The reverse approach is compared with the more traditional forward analysis, in which an expansion is derived that transforms the stimulus to a prediction of the spike train response. Important aspects of the application of these analytical approaches are considered. 3. All four interneurons were found to have identical frequency tuning, as assessed by the accuracy with which different frequency components of stimulus waveforms could be reconstructed with a linear expansion. The interneurons encoded significant information about stimulus frequencies between 5 and 80 Hz, which peak sensitivities at approximately 15 Hz. 4. All four interneurons were found to have identical stimulus/response latencies. The mean latency between a stimulus component and the corresponding elicited spike was 17 ms. All four interneurons also had identical integration times. The integration time, measured by the duration of stimulus, which could affect the probability of spiking, was approximately 50 ms. 5. The accuracy of the encoding can be expressed as a signal-to-noise ratio, where the noise is a scaled difference between the original signal and the best estimate of the signal. Peak signal-to noise ratios of approximately 1 were obtained for the cells across all stimulus power levels, using only the linear expansion term. Analysis of the data indicated that the consideration of second-order nonlinear transformations of the stimulus would not have increased the calculated encoding accuracy. 6. The encoding accuracy also can be expressed in the information theoretic units of bits/second, which characterizes the information transmission rate of the cell. Bits/second values varied between 10 and 80 for the 22 different cells in our experimental set. The information rate values were highly correlated with the mean spike rates of the interneurons, but were not correlated with the stimulus power levels. However, normalizing the absolute information rates by the mean spike rate in each case yielded a measure of bits/spike that was remarkably invariant across all experiments. The measured bits/spike rate was approximately 1 for all experiments. This result is discussed in the context of recent theoretical studies on optimal encoding. 7. Although the dynamic sensitivities of the four interneurons were identical, their directional sensitivities are known to be orthogonal. Thus the cells are complementary to one another from a functional standpoint: whereas a particular cell will be insensitive to air currents from some directions, one or more of the other three cells will be sensitive to stimuli from those directions... PMID- 8727383 TI - Information theoretic analysis of dynamical encoding by filiform mechanoreceptors in the cricket cercal system. AB - 1. The stimulus/response properties of 20 mechanosensory receptors in the cricket cercal sensory system were studied using electrophysiological techniques. These receptors innervated filiform hairs of various lengths and directional selectivities. Previous studies have characterized the sensitivity of such cells to the direction of air currents and to the amplitude of sinusoidal stimuli. In the experiments reported here, the quantity and quality of information encoded in the receptors' elicited responses about the dynamics of more complex air current waveforms were characterized. 2. Based on a white analysis of receptor response properties, the median frequency of each receptor's frequency tuning curve was found to be strongly correlated with the length of its associated mechanosensory hair. The receptors connected to hairs > 900 microns encoded frequencies below approximately 150 Hz very accurately and the receptors connected to shorter hairs encoded progressively higher bands of frequencies. These results were interpreted within the constraints imposed by the biomechanics of the air current-to-cercus boundary. 3. The encoding accuracy was expressed in the information theoretic units of bits/second, which characterizes the information transmission rate of a receptor. The information rates of the neuronal spike trains ranged from 75 to 220 bits/s. The information transmission rate was not correlated with the length of the mechanosensory hair. The average amount of information transmitted per action potential was negatively correlated with receptor hair length and ranged between 0.6 and 3.1 bits/spike. Decoding of the receptor responses was restricted to linear transformations of the spike trains. 4. The stimulus/response latencies of the different receptors ranged between 5 and 11 ms, and the integration time of the receptors ranged between 8 and 30 ms. The latency of a receptor was only weakly correlated with the length of its associated hair, and a receptor's integration time was correlated with hair length. 5. The stimulus/response phase difference for receptor cells that innervated hairs longer than approximately 800 microns increased with frequency > 50 Hz. The phase responses for receptor cells connected to hairs < 800 microns did not vary for frequencies > 50 Hz. PMID- 8727384 TI - Activation of GABAB receptors at individual release boutons of the crayfish opener neuromuscular junction produces presynaptic inhibition. AB - 1. Presynaptic inhibition in crustaceans involves the activation of gamma aminobutyric acid-A (GABAA) receptors that produce an increase in chloride conductance at excitatory axon terminals. Such inhibition produced by single inhibitory pulses is blocked by picrotoxin, a GABAA antagonist. 2. Presynaptic inhibition produced by bath application of GABA was not blocked by picrotoxin. Measurements of the membrane resistance of the excitatory axon terminals revealed that substantial presynaptic inhibition still persisted after 50 microM picrotoxin had completely blocked the increase in conductance produced by 10 microM GABA. 3. Baclofen, a GABAB agonist, reduced release from the excitatory nerve terminals, and 20H-Saclofen, a GABAB antagonist, blocked the effect of baclofen and the presynaptic inhibition produced by 10 microM GABA. 4. 20H Saclofen alone did not block presynaptic inhibition produced by 100 microM GABA, and the combined action of both 20H-Saclofen and picrotoxin was required to block such effects. 5. The excitatory nerve terminals seem to contain GABAA and GABAB receptors. The GABAB receptors are preferentially activated at lower GABA concentrations (in the microM range), whereas both the GABAA and GABAB receptors are activated at high GABA concentrations. PMID- 8727386 TI - Short-latency disparity vergence responses and their dependence on a prior saccadic eye movement. AB - 1. A dichoptic viewing arrangement was used to study the initial vergence eye movements elicited by brief horizontal disparity steps applied to large textured patterns in three rhesus monkeys. Disconjugate steps (range, 0.2-10.9 degrees) were applied to the patterns at selected times (range, 13-303 ms) after 10 degrees leftward saccades into the center of the pattern. The horizontal and vertical positions of both eyes were recorded with the electromagnetic search coil technique. 2. Without training or reinforcement, disparity steps of suitable amplitude consistently elicited vergence responses at short latencies. For example, with 1.8 degrees crossed-disparity steps applied 26 ms after the centering saccade, the mean latency of onset of convergence for each of the three monkeys was 52.2 +/- 3.8 (SD) ms, 52.3 +/- 5.2 ms, and 53.4 +/- 4.1 ms. 3. Experiments in which the disparity step was confined to only one eye indicated that each eye was not simply tracking the apparent motion that is saw. For example, when crossed-disparity steps were confined to the right eye (which saw leftward steps), the result was (binocular) convergence in which the left eye moved to the right even though that eye had seen only a stationary scene. This movement of the left eye cannot have resulted from independent monocular tracking and indicates that the vergences here derived from the binocular misalignment of the two retinal images. 4. The initial vergence responses to crossed-disparity steps had the following main features. 1) They were always in the correct (i.e., convergent) direction over the full range of stimuli tested, the initial vergence acceleration increasing progressively with increases in disparity until reaching a peak with steps of 1.4-2.4 degrees and declining thereafter to a nonzero asymptote as steps exceeded 5-7 degrees. 2) They showed transient postsaccadic enhancement whereby steps applied in the immediate wake of a saccadic eye movement resulted in much higher initial vergence accelerations than the same steps applied some time later. The response decline in the wake of a saccade was roughly exponential with time constants of 67 +/- 5 (SD) ms, 35 +/- 2 ms, and 54 +/- 4 ms for the three animals. 3) That the postsaccadic enhancement might have resulted in part from the visual stimulation associated with the prior saccade was suggested by the finding that enhancement could also be observed when the disparity steps were applied in the wake of (conjugate) saccadelike shifts of the textured pattern. However, this visual enhancement did not reach a peak unit 17 37 ms after the end of the "simulated" saccade, and the peak enhancement averaged only 45% of that after a "real" saccade. 4) Qualitatively similar transient enhancements in the wake of real and simulated saccades have also been reported for initial ocular following responses elicited by conjugate drifts of the visual scene. We replicated the enhancement effects on ocular following to allow a direct comparison with the enhancement effects on disparity vergence using the same animals and visual stimulus patterns and, despite some clear quantitative differences, we suggest that the enhancement effects share a similar etiology. 5. Initial vergence responses to uncrossed-disparity steps had the following main features. 1) They were in the correct (i.e., divergent) direction only for very small steps (< 1.5-2.5 degrees), and then only when postsaccadic delays were small; when the magnitude of the steps was increased beyond these levels, responses declined to zero and thereafter reversed direction, eventually reaching a nonzero (convergent) asymptote similar to that seen with large crossed disparity steps; convergent responses were also seen with larger vertical disparity steps, suggesting that they represent default responses to any disparity exceeding a few degrees. 2) As the postsaccadic delay was increased, responses to small steps (1.8 degrees) declined to zero and thereafter re PMID- 8727385 TI - Persistence of the olfactory receptor current in a wide variety of extracellular environments. AB - We measured the current activated by cytoplasmic adenosine 3':5'-cyclic monophosphate (cAMP) in olfactory cilia from the frog Rana pipiens. The odorant induced current in frog olfactory receptor neurons was also measured for comparison. In both cases, recordings were performed near the neuronal resting potential in a variety of extracellular bath solutions. 2. In Ca(2+)-free baths, cAMP activated an inward current in excised olfactory cilia that was carried entirely by cations. As extracellular Ca2+ was increased, the cationic current decreased while a second current, carried by C1-, increased. Total cAMP-activated current decreased with increasing extracellular CA2+. When external Na+ but not Ca2+ was eliminated, only the C1- component of the current persisted. When external Na+ and Ca2+ were both removed, there was no cAMP-activated current. 3. In receptor neurons, the total odorant-induced receptor current varied in a similar way with the extracellular ionic environment. Under conditions favoring the anionic receptor current, the response amplitude decreased and the latency increased. 4. It is known that olfactory receptor currents persist in a wide variety of extracellular environments. This persistence can be sufficiently explained by the balance between cationic and anionic currents demonstrated here. PMID- 8727387 TI - Response properties of units in the dorsal cochlear nucleus of unanesthetized decerebrate gerbil. AB - 1. The electrophysiological responses of single units in the dorsal cochlear nucleus of unanesthetized decerebrate Mongolian gerbil (Meriones unguiculatus) were recorded. Units were classified according to the response map scheme of Evans and Nelson as modified by Young and Brownell, Young and Voigt, and Shofner and Young. Type II units have a V-shaped excitatory response map similar to typical auditory nerve tuning curves but little or no spontaneous activity (SpAc < 2.5 spikes/s) and little or no response to noise. Type I/III units also have a V-shaped excitatory map and SpAc < 2.5 spikes/s, but have an excitatory response to noise. Type III units have a V-shaped excitatory map with inhibitory sidebands, SpAc > 2.5 spikes/s, and an excitatory response to noise. Type IV-T units typically also have a V-shaped excitatory map with inhibitory sidebands, but have a highly nonmonotonic rate versus level response to best frequency (BF) tones like type IV units, SpAc > 2.5 spikes/s, and an excitatory response to noise. Type IV units have a predominantly inhibitory response map above an island of excitation of BF, SpAc > 2.5 spikes/s, and an excitatory response to noise. We present results for 133 units recorded with glass micropipette electrodes. The purpose of this study was to establish a normative response map data base in this species for ongoing structure/function and correlation studies. 2. The major types of units (type II, type I/III, type III, type IV-T, and type IV) found in decerebrate cat are found in decerebrate gerbil. However, the percentage of type II (7.5%) and type IV (11.3%) units encountered are smaller and the percentage of type III (62.4%) units is larger in decerebrate gerbil than in decerebrate cat. In comparison, Shofner and Young found 18.5% type II units, 30.6% type IV units, and 23.1% type III units using metal electrodes. 3. Two new unit subtypes are described in gerbil: type III-i and type IV-i units. Type III-i units are similar to type III units except that type III-i units are inhibited by low levels of noise and excited by high levels of noise whereas type III units have strictly excitatory responses to noise. Type IV-i units are similar to type IV units except that type IV-i units are excited by low levels of noise and become inhibited by high levels of noise whereas type IV units have strictly excitatory responses to noise. Type III-i units are approximately 30% of the type III population and type IV-i units are approximately 50% of the type IV population. 4. On the basis of the paucity of classic type II units and the reciprocal responses to broadband noise of type III-i and type IV-i units, we postulate that some gerbil type III-i units are the same cell type and have similar synaptic connections as cat type II units. 5. Type II and type I/III units are distinguished from one another on the basis of both their relative noise response, rho, and the normalized slope of the BF tone rate versus level functions beyond the first maximum. Previously, type II units were defined to be those nonspontaneously active units with rho values < 0.3 where rho is defined as the ratio of the maximum noise response minus spontaneous rate to the maximum BF tone response minus spontaneous rate. In the gerbil, the average rho value for type II units is 0.25, although a few values are > 0.3, and the rate-level curves are consistently nonmonotonic with normalized slopes steeper than than -0.007/dB. The average rho value for type I/III units is 0.54, although a few values are < 0.3, and the rate-level curves tend to saturate with slopes shallower than 0.006/dB. In general, the response properties of type II units recorded in gerbil are similar to those recorded in decerebrate cat. 6. In comparison to decerebrate cat, the lower percentage of type IV units recorded in decerebrate gerbil may be due to a species difference (a reduced number of type II units in gerbil) or an electrode bias. PMID- 8727388 TI - Physiological properties of rat ventral pallidal neurons recorded intracellularly in vivo. AB - 1. The physiology of ventral pallidal (VP) cells was investigated using in vivo intracellular recording and staining techniques in adult rats. Based on electrophysiological criteria, three different types of cells were found: type A cells, which fired phasic spikes that did not exhibit a substantial afterhyperpolarization (AHP), type B cells, which exhibited a slow ramplike depolarization that preceded the short-duration action potential; the spike was followed by a prominent AHP, and type C cells, which were the only cells that fired spikes in couplets or bursts, with the spikes in a burst exhibiting a progressive increase in duration and a decrease in amplitude. These cells also exhibited a rebound low threshold spikelike event. Furthermore, 18% of the VP cells recorded exhibited a slow subthreshold oscillation of the membrane potential (< 1 Hz). 2. The response of VP cells to stimulation of fibers arising from the prefrontal cortex, nucleus accumbens, and mediodorsal thalamic nucleus (MD) was examined. In contrast to our initial predictions, all cells responded to nucleus accumbens stimulation with excitation. Type A and B cells responded to nucleus accumbens stimulation with excitation and to MD stimulation with antidromic-like responses, orthodromic excitation, or evoked inhibitory postsynaptic potentials. Only type A cells responded to prefrontal cortical stimulation. Type C cells only responded to stimulation of the nucleus accumbens, which resulted in evoked excitatory postsynaptic potentials. 3. The cells in the VP therefore can be segregated into three physiologically defined groups according to action potential discharge patterns and their response to afferent fiber stimulation. PMID- 8727389 TI - Cross-correlation analysis reveals laminar differences in thalamocortical interactions in the somatosensory system. AB - 1. Spontaneous and stimulus-induced activity were recorded from corresponding somatotopic representations in the ventroposterolateral nucleus (VPL) of the thalamus and primary somatosensory (SI) cortex of intact, halothane-anesthetized cats. Thalamic and cortical neurons with overlapping receptive fields on the hairy skin of the forelimb were excited by a series of interleaved air jets aimed at multiple skin sites. 2. The laminar locations of 68% (240 of 355) of the neurons recorded in SI cortex were histologically reconstructed and responses of these 240 SI neurons were analyzed with respect to responses recorded from 118 thalamic neurons. Maximum responsiveness during the initial onset (1st 100 ms) of air jet stimulation was similar for neurons distributed throughout all layers of SI cortex (2-4 spikes per stimulus) and did not differ significantly from VPL responses. During the subsequent plateau phase of the stimulus, VPL neurons discharged at a mean rate of 19.0 spikes/ s and neurons in cortical layers II, IIIa, IIIb, and IV discharged at similar rates. Mean responsiveness during the plateau phase of the stimulus was significantly reduced among neurons in cortical layers V and VI and only averaged 7.1 and 3.9 spikes/s, respectively. 3. Responses recorded simultaneously from pairs of thalamic and cortical neurons were analyzed with cross-correlation analysis to determine differences in the incidence and strength of neuronal interactions as a function of cortical layer. Among 421 thalamocortical neuron pairs displaying stimulus-induced responses, 68 neuron pairs exhibited significant interactions during air jet stimulation. A laminar analysis revealed that 28% (45 of 163) of the neurons in the middle cortical layers displayed significant interactions with thalamic neurons, whereas only 14% (13 of 92) of superficial layer neurons and 6% (10 of 166) of deep layer neurons were synchronized with thalamic activity during air jet stimulation. When thalamocortical efficacy for different layers of cortex was plotted as a cumulative frequency distribution, the strongest interactions in the middle cortical layers were twice as strong as interactions involving the superficial or deep cortical layers. 4. More than 70% of stimulus-induced interactions involved thalamic discharges followed by subsequent cortical discharges and the majority of these interactions involved interspike intervals of < or = 3 ms. Nearly 75% (27 of 37) of interactions in the thalamocortical direction that involved cortical neurons in layers IIIb and IV transpired within a 3-ms interspike interval. For interactions with superficial or deep cortical layers, the proportion of thalamocortical interactions transpiring within 3 ms was only 58% (7 of 12) and 33% (2 of 6), respectively. 5. Cross-correlation analysis of spontaneous activity indicated that 124 pairs of thalamic and cortical neurons displayed synchronous activity in the absence of sensory stimulation. A laminar analysis indicated that similar proportions of cortical neurons in each layer were synchronized with thalamic activity in the absence of cutaneous stimulation. Thus 27% (44 of 163) of middle layer neurons, 30% (28 of 92) of superficial layer neurons, and 31% (51 of 166) of deep layer neurons displayed spontaneous interactions with thalamic neurons. The temporal pattern of spontaneous activity was examined with autocorrelation analysis to determine whether neuronal oscillations were essential for coordinating thalamic and cortical activity in the absence of peripheral stimulation. Only 18.5% (23 of 124) of spontaneous interactions between thalamic and cortical neurons were associated with periodic activity, which suggests that thalamocortical synchronization occurs before the constituent neurons begin to oscillate. 6. The influence of sensory stimulation on spontaneous interactions was examined in 31 pairs of thalamic and cortical neurons that exhibited interactions during prestimulus and stimulus in PMID- 8727390 TI - Characterization of GABAA receptor function in human temporal cortical neurons. AB - 1. Surgically resected tissue from the tip of the human temporal lobe of seven patients undergoing temporal lobectomy was employed to study functional properties of GABAergic inhibition mediated through activation of GABAA receptors, using patch-clamp recording techniques in acutely isolated neurons and in slices of human temporal cortex. 2. Human temporal cortical pyramidal neurons from surgically resected tissue could be acutely isolated with the use of conventional methods. These neurons appeared normal in morphology, in their intrinsic membrane properties, and in their response to application of exogenous gamma-aminobutyric acid (GABA). 3. Application of GABA to acutely isolated human temporal cortical neurons elicited a large current with an average reversal potential of -65 mV, presumably mediated through a GABAA-activated chloride conductance. Application of varying concentrations of GABA generated a concentration/response relationship that could be well-fitted by a conventional sigmoidal curve, with an EC50 of 25.5 microM and a Hill coefficient of 1.0 4. Coapplication of the benzodiazepine clonazepam and 10 microM GABA augmented the amplitude of the GABA response. The concentration dependence of this benzodiazepine augmentation could be best-fitted by an equation assuming that the benzodiazepine interacted with two distinct binding sites, with differing potencies. The high-potency site had an EC50 of 0.06 nM and maximally contributed 38.5% augmentation to the total effect of clonazepam. The lower potency site had an EC50 of 16.4 nM, and contributed 66.1% maximal augmentation to the overall effect of clonazepam. These data derived from adult human temporal cortical neurons were very similar to our findings in adult rat sensory cortical neurons. 5. The effects of equimolar concentrations (100 nM) of clonazepam, a BZ1 and BZ2 agonist, and zolpidem, a selective BZ1 agonist, on acutely isolated human temporal cortical neurons were also investigated. Zolpidem and clonazepam were equally effective (71.5 vs. 65.0%, respectively) in potentiating GABA responses elicited by application of 10 microM GABA. This suggests that many of the functional benzodiazepine receptors in these neurons were of the BZ1 variety. 6. GABAergic synaptic inhibition was also studied with the use of patch-clamp recordings in slices of human temporal cortex. Extracellular stimulation at the white matter/gray matter border elicited compound synaptic events in layer II-V cortical neurons. These events usually consisted of an early excitatory postsynaptic potential (EPSP) and a late multiphasic inhibitory postsynaptic potential (IPSP). Application of either clonazepam or zolpidem (both at 100 nM) to the slice during extracellular stimulation reversibly augmented the late compound IPSP. 7. Spontaneous IPSPs were also recorded in approximately 50% of human temporal cortical neurons. These events did not have a preceding EPSP and were usually monopolar, with a single exponential rise and decay. This supported the idea that these events were triggered by spontaneous activity of GABAergic interneurons. Bath application of either clonazepam or zolpidem (both at 100nM) to the slice during ongoing spontaneous IPSP activity increased the amplitude and lengthened the time constant of decay of these events. 8. To our knowledge, this is one of the first detailed characterizations of the functional properties of GABAA-mediated inhibition in human cortical neurons using patch-clamp recordings in both isolated cells and slices of resected temporal cortex. Isolated pyramidal neurons exhibited GABAA-mediated currents that were comparable in many aspects with GABA currents recorded from adult rat cortical neurons, including similar GABA concentration/response curves, and similar two differing potency site effects for clonazepam augmentation of GABA currents. In addition, evoked and spontaneous IPSPs recorded in human cortical neurons appeared similar to IPSPs in rat cortical PMID- 8727391 TI - Spatiotemporal characteristics of 5-HT and dopamine-induced rhythmic hindlimb activity in the in vitro neonatal rat. AB - 1. Rhythmic activity was induced with either serotonin (5-HT; 10-100 microM) or dopamine (0.1-1.0 mM), in the in vitro spinal cord preparation of neonatal rats, with one intact hindlimb attached. Patterns of activity were investigated with multiple EMG recordings and the spatiotemporal characteristics of 5-HT- and dopamine-induced activity compared. 2. Dopamine-induced rhythmic activity was slow (cycle duration: 2.2-70.1 s) and irregular, whereas rhythmic activity induced by 5-HT was fast (cycle duration: 1.3-5.1 s) and regular. 3. During 5-HT- and dopamine-induced rhythmic activity, the timing of muscular activity was similar for hip flexors and hip adductors, for semimembranosus (hip extensor), and for muscles controlling the ankle and the foot. 4. In contrast, notable differences in the phase in the pattern induced by 5-HT compared with that induced by dopamine were found in the biceps femoris, semitendinosus, and quadriceps muscles. Biceps femoris and semitendinosus (functional hip extensors and knee flexors) were always extensor-like during 5-HT-induced activity, whereas in dopamine, these muscles displayed flexor-like bursts and double bursts as well as extensor-like bursts. Lack of EMG activity in biceps femoris and semitendinosus was encountered also in dopamine. In rectus femoris, vastus lateralis, and vastus medialis (main function: knee extension), the activity was dominated by flexor-like bursts in 5-HT, whereas in dopamine the activity was shifted to a predominantly extensor-like pattern. 5. The relationship between flexor and extensor burst duration and cycle duration was more variable than described for locomotor activity in adult animals. 6. The relative timing of muscle activity was stable from P0 to P4. The most important difference between rats aged 0-1 days and rats aged 2-4 days was a delayed flexor-extensor transition in older animals. 7. The complex timing of hindlimb muscle activity was relatively unchanged after transecting all dorsal roots. 8. Finally, the relationship between flexor and extensor activity and ventral root discharges was determined. It was found that the L2 ventral root burst was in phase with simple flexors while the L5 burst coincide with the extensor phase. 9. We conclude, that 5-HT and dopamine can activate spinal central pattern generators (CPGs) that already at birth are able to produce distinct patterns of motor activity. Modulatory inputs thus seems to be able to reconfigure the CPGs to produce specific motor outputs. PMID- 8727392 TI - Retinal slip neurons in the nucleus of the optic tract and dorsal terminal nucleus in cats with congenital strabismus. AB - 1. Visual response properties of neurons in the nucleus of the optic tract (NOT) and dorsal terminal nucleus of the accessory optic tract (DTN) were electrophysiologically investigated in five congenitally strabismic cats and compared with normal adult cats and 3- to 4-wk-old kittens. 2. As in normal cats, NOT-DTN cells of strabismic cats preferred horizontal ipsiversive stimulus movement. However, NOT-DTN neurons in strabismic cats altered their activity to a lesser amount per degree change of stimulus direction than do normal adult cats. In addition, NOT-DTN cells in strabismic cats exhibited a broader directional tuning, i.e., they increase their activity to a broader range of directions than control NOT-DTN cells. 3. Spontaneous activity and activity difference between preferred and nonpreferred direction were significantly lower in NOT-DTN neurons of strabismic cats than in normal adult cats and resembled that found in 3-wk-old kittens. Maximal stimulus-related activity was lower than in normal adult cats but higher than in kittens. 4. Visual latencies to onset of movement in the preferred direction were indistinguishable in strabismic and in normal adult cats. Visual latencies to onset of movement in the nonpreferred direction, however, were shorter in strabismic cats than in normal adult cats. 5. The average velocity tuning curve of NOT-DTN cells in strabismic cats was very flat without a well-defined optimal stimulus velocity. Thus it closely resembled data from 3-wk-old kittens. 6. Binocular convergence was significantly altered to a stronger dominance of the contralateral eye in NOT-DTN of strabismic cats. This reduction of binocular neurons was less pronounced than in cats with artificially induced strabismus or in 3-wk-old kittens. 7. In conclusion, the data presented here for retinal slip neurons in the NOT-DTN of strabismic cats closely resemble those from 3-wk-old kittens where no functional cortical input to the NOT-DTN is present. However, the elevated stimulus-driven activity and the still relatively high degree of binocularity give a clear indication of a functional, albeit weak and abnormal, cortical input to the NOT-DTN in these naturally strabismic cats. PMID- 8727393 TI - Optokinetic nystagmus in cats with congenital strabismus. AB - 1. Eye movements were recorded in seven innately esotropic cats during monocular and binocular horizontal optokinetic stimulation, using the search coil technique in five cats and electrooculography in two cats. 2. During closed loop measurements in these strabismic cats, slow phases of optokinetic nystagmus (OKN) were characterized by an overall reduced gain when compared with normal controls. In addition, response gain to monocular nasotemporal stimulation was even more reduced than that to temporonasal stimulation, resulting in an increased asymmetry of closed loop gain. 3. During open loop measurements, eye velocity in strabismic cats was very low at all velocities tested. 4. Differential analysis of the symmetry of OKN revealed that all our strabismic cats had a "good" or more symmetric and a "poor" or more asymmetric eye. In addition, when analyzed separately at individual velocities, the symmetry index of the good eye was fairly constant over the velocity range tested. By contrast, the symmetry index of the poor eye dropped dramatically at higher stimulus velocities. 5. To analyze the relationship of OKN symmetry and cortical physiology, we calculated the ratio between the percentage of neurons driven by one eye in the ipsilateral and the contralateral cortical hemisphere. We found a weak correlation between OKN symmetry and this cortical symmetry index (P < 0.05, analysis of variance). 6. In conclusion, slow eye movements in cats with congenital esotropia are characterized by extremely low gain, especially at higher stimulus velocities. In addition, OKN symmetry during monocular stimulation is decreased. Our data suggest that OKN symmetry is weakly correlated with the proportion of binocular neurons in the visual cortex ipsilateral to the stimulated eye. However, OKN characteristics seem to reflect to a higher degree the response properties of neurons in the pretectal nucleus of the optic tract and the dorsal terminal nucleus of the accessory optic system than properties of neurons in the visual cortex. PMID- 8727394 TI - Capsaicin-induced currents with distinct desensitization and Ca2+ dependence in rat trigeminal ganglion cells. AB - 1. Whole cell patch-clamp records from cultured rat trigeminal ganglion cells having soma diameters ranging from 20 to 50 microM revealed that capsaicin activated two inward currents and an outward current. At -60 mV, the inward currents could be distinguished by their different peak times, which were 4.2 +/- 3.1 and 41.4 +/- 16.4 (SD) s. 2. Cells with the smallest soma diameters had the largest current densities. 3. The more rapidly activating current had a linear current-voltage relation and a reversal potential near 0 mV. 4. The more slowly activating current is not a Ca(2+)-activated Cl- current. 5. The peak of the rapid current (Ip)-capsaicin concentration (C) relationship was characterized by Ip/Ipmax = [1 + (C/Kd)n]-1, where n = 1.2 and the dissociation constant (Kd) = 0.68 microM. 6. The rapidly activating current was heterogeneous in regards to both its rate of activation and extent of desensitization. In cells bathed in buffer containing calcium and held at -60 mV, most of the capsaicin-activated currents desensitized. Removal of extracellular Ca2+ could reduce, eliminate, or have no effect on desensitization. 7. At positive holding potentials the currents very slowly desensitized, even in the presence of Ca2+. 8. Repeated 30-s applications of 1 microM capsaicin separated by 0.5, 2.5, and 5.5 min all induced tachyphylaxis. Tachyphylaxis decreased exponentially until the current remained approximately constant. Decreasing the time between capsaicin applications increased the extent of tachyphylaxis, whereas elimination of extracellular Ca2+ markedly reduced tachyphylaxis. PMID- 8727395 TI - Model circuit of spiking neurons generating directional selectivity in simple cells. AB - 1. We here consider the property of directional selectivity (DS) in simple cells of layer 4 of cat area 17 as an instance of a receptive field (RF) transformation between two monosynaptically connected neuron populations: the afferent geniculate (lateral geniculate nucleus, LGN) cells and their target, layer 4 simple cells. We have studied this particular RF transformation because the large set of experimental data available allowed us to restrain the synaptic organization of our model layer 4 circuitry. 2. The one-compartment, spiking model neurons of the layer 4 circuitry are excitatory (adapting) or inhibitory (nonadapting). They all have simple-cell RFs composed of two spatially separated ON and OFF subregions. The sequence of the subregions across the neurons' RFs, which is determined by the geniculocortical inputs they receive, varies independently from their preferred direction of stimulus motion, which is determined by spatial asymmetries in their corticocortical inputs. 3. Synaptic transmission in the model layer 4 circuitry is mediated via non-N-methyl-D aspartate (non-NMDA) receptors (geniculocortical excitation), via NMDA receptors (corticocortical excitation), and via gamma-aminobutyric acid-A receptors (corticocortical inhibition). Excitatory and inhibitory cortical neurons receive the same afferents. However, excitatory neurons form efferent synapses exclusively with neurons having the same RF characteristics, and preferentially with those having the same RF position. Inhibitory neurons form synapses preferentially with neurons having different RF characteristics or adjacent RF positions. 4. By comparing the neurons' numerically computed responses to visual stimuli with those of actual simple cells, the topology of the corticocortical connections has been constrained. The experimental responses to stationary and moving, and to bar as well as grating, stimuli are consistently reproduced with a single constant parameter setting. 5. Subsequently, the model has been analyzed from a system-theoretic approach and has been manipulated in order to find the components critical for its proper functioning. Variations on the model have been simulated for evaluating the performance of alternative connection schemes. 6. Spatially opponent inhibition between model simple cells with antagonistic RF subregions is necessary for the restoration of linearity lost at the LGN output. It hyperpolarizes model simple cells when the contrast polarity of an efficient stimulus is reversed and prevents, particularly in directionally nonselective cells, a frequency doubling of the responses to sine wave gratings of low spatial frequencies. 7. Directionally opponent inhibition between model simple cells preferring opposite directions of motion is necessary for the generation of genuine DS (a ratio of firing rates > 2 for opposite directions of motion). 8. The corticocortical excitatory polysynaptic feedback loops in the model are able to provide the time delays needed to generate DS, and even to preserve DS at very low speeds. The strength spatial extension, and time course of this corticocortical feedback excitation, together with the dynamics of the geniculate afferents and the width of the RF, determine the tuning of model simple cells in the temporal and velocity domain. 9. The present model generates directionally selective responses to stimulus motion over distances smaller than the width of a single subregion and as small as the spacing between the afferent geniculate RFs. The direction-selective mechanism acts uniformly across the entire width of a subregion. Thus the position invariance of DS arises in the present model at the same level as DS itself. The same holds for the stimulus polarity (light vs. dark) invariance of DS. Consequently, there is no need for highly hierarchical models in which all these characteristics accumulate in simple cells by pooling from lower-order subunits or neurons. PMID- 8727396 TI - Central mesencephalic reticular formation (cMRF) neurons discharging before and during eye movements. AB - 1. One hundred twenty neurons were recorded in the central mesencephalic reticular formation (cMRF) of four rhesus monkeys, trained to make visually guided and targeted saccadic eye movements. Eye movements were recorded with the head fixed, using electrooculography (EOG) or subconjunctival scleral search coils. Seventy-six percent (92/120) of cells discharged before and during contraversive visually guided or targeted rapid eye movements, and 76% of these (70/92) responded during contraversive spontaneous saccades in the dark. cMRF neurons had large contraversive movement fields and either a high (> 10 spikes/s) or low background level of spontaneous activity in the dark. The optimal movement vectors (i.e., saccades with greatest response) were predominantly horizontal, although many had a vertical component. Cells with optimal movement vectors within +/- 25 degrees of pure vertical were more rostral in the MRF and were excluded from the analysis. 2. A subgroup of cMRF neurons (31 of 92) that discharged before and during visually guided saccades were examined for visual sensitivity. Slightly less than one-half of these cells (42%, 13/31) were visuomotor units, i.e., they responded to visual targets in the absence of eye movement. The other 58% (n = 18) did not discharge during the visual probe trial; they were movement-related cells. 3. Microstimulation (threshold 40-60 microA at 333 Hz) at the sites of many of these cMRF neurons produced contraversive saccadic eye movements at short latency (< 40 ms). The amplitude and direction of the elicited saccades were similar to the optimal movement vector determined from single-unit recording. This suggested that cMRF cells recorded at the same locus of electrical microstimulation participated in the network responsible for the production and control of rapid eye movements. 4. The 92 saccade-related neurons were divided into two groups on the basis of their background discharge rate. Firing rates for both low background (28%, n = 26) and high background (72%, n = 66) cells increased approximately 30 ms before contraversive saccades and reached a peak discharge just before saccade onset. The low background neurons had either no activity or generated a few spikes just before the end of ipsiversive saccades. The steady rate of discharge (> 10 spikes/s) of high background neurons was inhibited from approximately 20 ms before ipsiversive saccades until just before saccade end. 5. Cells were also subdivided on the basis of how their discharge rates fell at the end of saccades. Clipped cells (38%, n = 35) had activity that fell sharply with saccade offset. Partially clipped cells (62%, n = 57) had persistent firing in the 100 ms following the saccade that was > 20% higher than the firing during the 100 ms before the saccade. 6. Latencies between the 90% point on the rising edge of the peak discharge and the start of the saccade were < or = 5.3 ms for eye movement-related cells in two monkeys. Longer latencies (11-19 ms) were found when measured between the 10% point on the rising edge of the peak discharge and saccade onset. These latencies were equal to or shorter than those obtained for eye movement-related burst neurons in the intermediate and deep layers of the superior colliculus analyzed similarly. Delays between the peak discharge and peak eye velocity were 13.6-15.1 ms for the same group of cMRF eye movement-related cells. These were significantly shorter than the delays measured for eye movement neurons in the superior colliculus (SC) of one of the monkeys. These findings suggest that the buildup discharge of cMRF neurons occurs early enough before saccades to contribute to saccade triggering. The peak discharge, however, occurs with or after the burst in the SC, suggesting that this portion of the discharge serves a function other than saccade triggering. 7. The number of spikes in bursts associated with eye movement was correlated with saccade parameters. PMID- 8727397 TI - Spontaneous GABAA receptor-mediated inhibitory currents in adult rat somatosensory cortex. AB - 1. Spontaneous inhibitory synaptic currents (sIPSCs) were studied with whole cell voltage-clamp recordings from 131 pyramidal cells in adult rat somatosensory cortical slices. Neurons were intracellulary labeled with biocytin and classified as supragranular (SG, layers 2-3), layer IV (IV), or infragranular (IG, layer V) on the basis of the laminar localization of their somata. Somatic areas were similar for SG, IV, and IG neurons. All identified pyramidal cells generated high frequency gamma-aminobutyric acid (GABAA) receptor-mediated synaptic events. 2. Bath application of bicuculline blocked the sIPSCs and resulted in a decrease of approximately 0.5 nS in resting conductance and an inward shift in baseline current. 3. sIPSC frequency was significantly lower in SG versus IG or IV neurons, and this difference was accounted for by the occurrence of a higher percentage of bursts of sIPSCs in the IG and IV neurons. 4. Bath application of the alpha-amino-3-hydroxy-5-methyl-4-isoxazoleproprionic (AMPA) receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) decreased the frequency of sIPSCs by 13-21%. By contrast, application of the N-methyl-D-aspartate (NMDA) receptor antagonist D-2-amino-5-phosphonovaleric acid (D-AP5) generally had no effect on spontaneous IPSC frequency, suggesting that AMPA rather than NMDA receptor activation contributed to resting discharge of inhibitory interneurons. 5. Addition of tetrodotoxin (TTX) to the perfusion medium reduced the spontaneous IPSC frequency by approximately 30-55%. The miniature IPSCs (mIPSCs) seen in TTX containing solutions had a frequency of approximately 10 Hz and an average conductance of 0.42-0.48 nS. 6. The kinetic properties of mIPSCs generated in pyramidal cells of different layers were the same, with the rise times of approximately 0.9 ms and decay time constants of approximately 8 ms at a holding potential of 0 mV. The decay phase of mIPSCs was generally fitted by one exponential and displayed a voltage dependence with an e-fold increase in decay time constant for a every 198-mV depolarization. 7. These results show that there is ongoing spontaneous release of GABA in neocortical slices that gives rise to high-frequency impulse-related and non-impulse-related postsynaptic inhibitory currents. Activation of AMPA receptors on inhibitory interneurons accounts for only a small proportion of the GABAA receptor-mediated events. Judging from the distribution of mIPSC frequencies in neurons of different laminae, there is a relatively uniform distribution of inhibitory synapses throughout the cortex. Tonic activation of GABAA receptors on neocortical pyramidal neurons generates an increase in resting membrane conductance that may play an important role in vivo by preventing the development of hyperexcitability, modulating excitatory synaptic events, and controlling the rate and patterns of spike discharge. PMID- 8727398 TI - Electrophysiological mapping of GABAA receptor-mediated inhibition in adult rat somatosensory cortex. AB - 1. gamma-Aminobutyric acid-A (GABAA) receptor-mediated synaptic currents evoked by intracortical stimulation in rat somatosensory cortical slices maintained in vitro were studied using the whole cell patch-clamp technique. All anatomically identified pyramidal neurons of layer II-III (SG neurons), layer IV (IV neurons), and layer V (IG neurons) generated evoked inhibitory postsynaptic currents (eIPSCs) that were blocked by bicuculline. At threshold, eIPSCs had kinetic properties (rise time of 0.9 ms and decay time constant of 9 ms) similar to those of spontaneous IPSCs generated in the same cells. 2. The strength of inhibition was quantified by determining the stimulus threshold for evoking responses and the relationship between stimulus strength and eIPSC peak amplitudes (input/output curve). For eIPSCs recorded in control solution, the input/output curve was about four times steeper than for eIPSCs recorded in the presence of the ionotropic glutamate receptor antagonists 6-cyano-7-nitroquinoxaline-2,3 dione (CNQX) and D-2-amino-5-phosphonovalerate (D-AP5), suggesting the dependence of GABAA inhibition on synpatic excitation of interneurons. 3. In the presence of CNQX and D-AP5, monosynaptic IPSCs, evoked by stimulation close to the recording patch pipette, had similar input/output curves in SG and IG neurons. This suggests that the level of monosynaptic inhibition generated in these two populations of cells is similar. 4. When the stimulus was moved to a distant site > 350 microns from the recorded neuron, either in vertical or in horizontal direction, the stimulus intensity required for evoking IPSCs was higher, and the input/output curve was less steep. This suggests that the density of GABAergic somata and axons projecting to the recorded neuron is lower at these distances than at more proximal sites. 5. The maximum horizontal distance over which IPSCs could be evoked ("horizontal field") was larger in layer V than in other layers. The horizontal field (distance between stimulating and recording pipettes) was 600 microns in layer II-III, 580 microns in layer IV, and 720 microns in layer V. Anatomic identification of the somatosensory cortical barrels indicated that the extent of GABAergic projections was larger than the barrel hollow and might thus form a substrate for interbarrel inhibition in layer IV during cross-wisker stimulation. 6. The maximum vertical inhibitory field was larger than the maximum horizontal field. IPSCs could be evoked in layer V neurons by layer I stimuli, showing that a powerful interlaminar inhibition is present that may play a role in synchronizing the activity of neurons in a column. IPSCs evoked by layer I stimulation frequently had slower kinetics than those elicited by stimulation at sites close to the soma. 7. These findings suggest that functional GABAergic projections are characterized by a large degree of convergence. Quantification of GABAA-mediated IPSCs indicates that this zone of inhibitory synaptic convergence onto a given pyramidal neuron is subdivided into a powerful local inhibitory zone and a surrounding area of long-range, less effective, inhibitory projections. Potential roles for these concentric inhibitory areas in cortical processing of sensory information are discussed. PMID- 8727399 TI - Additivity of loud-sound--induced threshold losses in the cat under conditions of active or inactive cochlear efferent-mediated protection. AB - 1. An additivity model for the accretion of cochlear sensorineural hearing losses has been described from studies in the guinea pig cochlea. Among other aspects, the model allows determination of how residual hearing losses after an initial exposure (E1) affect hearing losses to be expected to a subsequent second exposure (E2). In the present study, the model was applied to temporary hearing losses produced in the cat cochlea by loud pure tones at a frequency from 3 to 15 kHz, affecting regions from 2 to 28 kHz. Successive identical exposures, generally with an interexposure interval of approximately equal to 35 min, were used to produce compound action potential (CAP) threshold losses. Total losses after E2 were compared with those predicted by the model. Testing was carried out under conditions where olivocochlear bundle (OCB)-mediated protection was or was not activated. (As shown elsewhere, OCB-mediated protection is activated by particular binaural exposures, but not monaural exposure, and reduces threshold losses in the binaural condition with intact OCB compared with losses in either the monaural condition, or the binaural condition where the OCB was cut before loud sound.) 2. The additivity model was a very good predictor of total losses under a variety of conditions; different exposure frequencies, monaural and binaural exposures, and with intact or cut OCB pathways. In these exposures, the model's application could be generalized so that as long as residual losses just pre-E2 were well specified in an animal, total losses could be as well predicted using normative data bases of a single exposure with the same parameters. 3. The model also allowed determination of whether OCB-mediated protection was exercised during E2 in dual identical exposures. Expression of protection for E2 depended on whether E1 elicited protection. When tested with monaural (at 7 or 15 kHz) or binaural exposures (at kHz) for which E1 did not elicit protection, neither did E2. However, when tested with a binaural E1 (at 7, 11, or 15 kHz), which activated protection, E2 also elicited protection. In the latter case, for 7- and 11-kHz exposures, the amount of E2 protection increased with total hearing loss, a relationship similar to that seen for single exposures in cat and guinea pig. For 15-kHz exposure, the amount of E2 protection was constant across test frequencies. 4. Finally, a critical observation with 11-kHz exposure was that a binaural E1 eliciting protection was able to "prime" the OCB so that protection could be elicited by a subsequent monaural E2, which, by itself as a singel exposure, does not evoke protection. This result has important implications in terms of the physiology of the protective OCB pathways and clinically in terms of the manner in which loud-sound-induced hearing loss accumulated. PMID- 8727400 TI - Influence of instruction, prediction, and afferent sensory information on the postural organization of step initiation. AB - 1. Our previous study showed that two distinct postural modifications occurred when subjects were instructed to step, rather than maintain stance, in response to a backward surface translation: 1) the automatic postural responses to the surfaces perturbation were reduced in magnitude and 2) the anticipatory postural adjustments promoting foot-off were shortened in duration. This study investigates the extent to which task instruction, prediction of perturbation velocity, and afferent sensory information related to perturbation velocity are responsible for these postural modification. 2. Eleven human subjects were instructed in advance, to either maintain stance or step forward in response to a backward surface translation. Four different velocities of translation were used to perturb equilibrium. To assess the influence of predicted versus actual velocity information, the surface translations were presented in both a blocked order of increasing perturbation velocity (predictable) and a random order (unpredictable). Lower-extremity electromyographs (EMGs), ground reaction forces, and movement kinematics were quantified for both the automatic postural responses to perturbation and the anticipatory postural adjustments for step initiation. 3. The instruction to step was not solely responsible for the suppression of the automatic postural response. Prediction of perturbation velocity was required for significant suppression of the early automatic postural response when subjects stepped in response to the perturbation. When compared with the stance condition, the magnitude of the initial 50 ms of the automatic response in bilateral soleus and the left limb gastrocnemius (initial stance limb) was significantly reduced only when the perturbation velocities were presented in a blocked order. The magnitude of the automatic response was not reduced in the gastrocnemius of the right limb, which was always the initial swing limb and recruited for heel-off in the step conditions. This asymmetrical reduction of the gastrocnemius suggests that modification of the response was specific to the instruction, rather than a general decrease in the extensor muscle excitability. 4. The suppression of the early automatic postural response involved a change in the bias of the response. Despite the reduced magnitude during the predictable velocity step condition, the slope (i.e., gain) of the response with increasing velocities was not different from that of the stance condition. Thus the excitability of the automatic response was reduced by a relatively constant amount for each velocity when the perturbation velocity was predictable. 5. In contrast to the importance of velocity prediction for modification of the automatic postural response, actual velocity information was used for modification of the anticipatory postural adjustments when step was initiated in response to the surface perturbation. Regardless of whether the perturbation velocities were presented in a blocked or random order, the anticipatory postural adjustments were rapidly initiated and the duration of the postural adjustments for step initiation was shortened as the velocity of perturbation increased. 6. We conclude that the CNS uses prediction of perturbation velocity to modify the excitability of early automatic postural responses when the postural goal changes. In contrast, actual afferent velocity information can be used to modify the duration of the anticipatory postural adjustments for a voluntary step in response to perturbation. Thus the CNS utilizes feed-forward prediction to modify peripherally triggered postural responses, and utilizes immediate afferent information to modify the centrally initiated postural adjustments associated with voluntary movement. PMID- 8727401 TI - Motor-unit behavior in humans during fatiguing arm movements. AB - 1. The activity of 40 triceps brachii motor units was recorded from the dominant arms of 9 healthy adult volunteers (age 27.8 +/- 4.4 yr, mean +/- SD) during a fatigue task that included both isometric and anisometric contractions. The fatigue task lasted 8.3 min and consisted of 50 extension and 50 flexion movements of the elbow. Each movement (40 degrees in 0.8s) was separated by an isometric contraction. A constant load resisting extension of 17.7 +/- 3.0% of maximal voluntary contractions (MVC) was applied throughout the task. This paradigm enabled the direct contrast of motor-unit discharge behavior during the different types of fatiguing contractions. 2. Motor-unit behavior was examined to determine the relative contribution of two mechanisms for optimizing force production under fatiguing conditions: recruitment of motor units and modulation of motor-unit discharge following recruitment. Threshold torques for motor-unit recruitment thresholds were determined by ramp-and-hold isometric contractions. Motor-unit discharge was evaluated during the fatigue task by contrasting the number of motor-unit potentials (spikes) per contraction for concentric eccentric, and isometric contractions. 3. The fatigue task resulted in a 30 +/- 12% decline in the mean MVC of elbow extension. Recruitment of nine new motor units (23%) was evident during the fatiguing extension movements, often within five to seven movements (i.e., within 25-35 s). Each newly recruited motor unit had the largest recruitment threshold torque in that experiment. 4. Analysis of the motor units that were active from the beginning of the fatigue task revealed that the mean number of motor-unit spikes per contraction increased, or remained constant as fatigue ensued, yet for the majority of motor units it increased or remained constant. None of the newly recruited motor units demonstrated decreased number of mean spikes per contraction after recruitment. Further, concurrently active motor units displayed different discharge behavior in two-thirds of the subjects. It is proposed that if the neural drive to the muscle is distributed uniformly upon the motoneuron pool, peripheral feedback from the exercising muscle may modulate specific motoneuron discharge levels during fatigue. PMID- 8727402 TI - Reducing human biceps brachii spinal stretch reflex magnitude. AB - 1. Nine subjects received 6 baseline and 24 training sessions, each consisting of 250 elbow joint stretch perturbations into extension. The training sessions were designed to reduce the subjects' biceps brachii spinal stretch reflexes (SSR). Changes in longer-latency responses and short-latency brachioradialis responses were also monitored. Background electromyogram activity was recorded from the lateral head of triceps brachii during the biceps sampling intervals. These data were compared with those form 12 control subjects who received equal numbers of stretches over an extended baseline (i.e., without application of a training paradigm). 2. Training subjects reduced their mean biceps SSR responses by 24%, whereas control subjects increased their responses by 12% When changes in activity were grouped by sets of six consecutive sessions following the baseline interval, group-by-time interaction was observed. Training subjects showed significant reductions from baseline after the first raining set. The increased magnitude of biceps SSR for control subjects was significantly larger than baseline in sets 2-5. After the first training interval, all subsequent differences between groups were statistically significant. 3. The brachioradialis showed greater response variability, but these responses paralleled those seen in biceps brachii. The training group reduced their brachioradialis responses by 18%, whereas the control group increased their responses by 12%. Background activity recorded over the lateral head of triceps brachii during the biceps brachii SSR window became smaller in both groups. 4. Comparison of data between control subjects and subjects who had undergone biceps SSR up-training in previous studies suggests that the small increase seen among the present control subjects was probably not the result of a direct training effect. 5. Baseline differences in longer-latency response onset time were seen between groups. Over extended baseline sessions, longer-latency responses showed only a 1% onset, representing a change of 0.3 ms, in the control group. A 1.3% later onset, equaling a change of 1.1 ms, was observed over conditioning sessions in the training group. When analyzed by sets following baseline, neither group showed significant within- or between-group changes over time. 6. The magnitude of the longer-latency biceps brachii response showed 7% and 37% reductions for the control and training groups, respectively. No difference in set averages was seen within groups, but a significant difference was measured over sets 2-4 between groups. Reductions in the magnitude of longer-latency responses in training subjects were more obvious in later training sessions. 7. These data suggest that 1) the biceps brachii SSR can be reduced in human subjects; 2) concurrent changes are often observed in the synergist brachioradialis; and 3) the reduced biceps SSR magnitude is not linked to a compensatory increase in antagonist muscle activity. Reductions in the magnitude of longer-latency biceps brachii activity seen in parallel with the decreased SSR may imply that some degree of supraspinal processing is required to achieve this task. PMID- 8727403 TI - Temporal features of directional tuning by spinocerebellar neurons: relation to limb geometry. AB - 1. We showed previously that neurons in the dorsal spinocerebellar tract (DSCT) may encode whole-limb parameters of movement and posture rather than localized proprioceptive information. Neurons were found to respond to hindlimb movements in the sagittal plane with maximum activity for foot placements in one direction and minimum activity for placements in the opposite direction. In contrast, movement direction is not specifically encoded by response activity when movement are restricted to a single joint. 2. We now describe the spatiotemporal characteristics of DSCT directional sensitivity for the responses of 267 neurons to small amplitude (0.5 cm) perturbations of the cat hindlimb. A small platform attached to the left hind foot was perturbed along four or eight directions in the sagittal plane, eliciting significant responses in 261 (98%) of the cells. The responses typically consisted of a sequence of peaks and troughs in poststimulus spike density lasting 150 ms or more following limb perturbation. 3. Peaks of activity in particular poststimulus intervals were broadly tuned for the direction of the perturbation, as determined by fitting the firing rates recorded in response to each perturbation direction to a cosine model. The parameters of the cosine model, namely the amplitude of modulation, the direction of maximum response, and the goodness of fit to the model, were computed for each 4 ms poststimulus interval. The parameters all showed the same tendency to wax and wane with respect to poststimulus time. For each period during which the cell activity was highly correlated with tuning model, the tuning indicated a different best direction. Thus each cell's directional tuning could be characterized by a set of tuning maxima associated with specific poststimulus times, when the amplitude of the tuning reached a local maximum and the fit to the cosine model was highly significant (R2 > 0.85). 4. Directions of the tuning maxima for the total population of cells were not uniformly distributed within particular poststimulus intervals. There was a statistically significant directional bias for upward directed perturbations in the poststimulus interval between 20 and 40 ms, followed by a period of downward bias from 45 to 55 ms. Between 60 and 85 ms, the distribution of tuning maxima was significantly skewed backward, whereas a very strong bias for the forward direction was present at about 100 ms. 5. Because the tuning was determined from responses to a very small perturbations of the limb in a given posture, it was not clear whether the responses were related to specific joint angles or muscle lengths, or whether they somehow represented the kinematics of the whole limb. To address this point, we examined the responses of 95 cells in two animals that were each tested in two different limb positions. One position was an approximation of the normal standing position. The other position consisted of a shortening of the limb axis (with major changes in all joint angles) in one animal, or a rotation of the limb axis backward (with little change in joint angles) in the other. 6. We compared each cell's responses to the same perturbations applied in the two limb positions and found they could be identical, scaled in time or magnitude, or completely different in the two positions. A greater percentage of cells with different responses was found in the experiment with the limb axis rotated. In the other experiment, in which there were major differences in joint angles in the two positions, the responses were mostly the same or scaled in time in the two positions. We also determined the population directional biases for the two positions in each experiment, and found that phase differences between the vectors representing population biases for the two positions were minimized when they were measured relative to the orientation of the limb axis (limb coordinates) rather than to the extrinsic vertical (lab coordinates). 7. PMID- 8727404 TI - Olfactory neuronal responses in the primate orbitofrontal cortex: analysis in an olfactory discrimination task. AB - 1. The primate orbitofrontal cortex receives inputs from the primary olfactory (pyriform) cortex and also from the primary taste cortex. To investigate how olfactory information is encoded in the orbitofrontal cortex, the responses of single neurons in the orbitofrontal cortex and surrounding areas were recorded during the performance of an olfactory discrimination task. In the task, the delivery of one of eight different odors indicated that the monkey could lick to obtain a taste of sucrose. If one of two other odors was delivered from the olfactometer, the monkey had to refrain from licking, otherwise he received a taste of saline. 2. Of the 1,580 neurons recorded in the orbitofrontal cortex, 3.1% (48) had olfactory responses and 34 (2.2%) responded differently to the different odors in the task. The neurons responded with a typical latency of 180 ms from the onset of odorant delivery. 3. Of the olfactory neurons with differential responses in the task, 35% responded solely on the basis of the taste reward association of the odorants. Such neurons responded either to all the rewarded stimuli, and none of the saline-associated stimuli, or vice versa. 4. The remaining 65% of these neurons showed differential selectivity for the stimuli based on the odor quality and not on the taste reward association of the odor. 5. The findings show that the olfactory representation within the orbitofrontal cortex reflects for some neurons (65%) which odor is present independently of its association with taste reward, and that for other neurons (35%), the olfactory response reflects (and encodes) the taste association of the odor. The additional finding that some of the odor-responsive neurons were also responsive to taste stimuli supports the hypothesis that odor-taste association learning at the level of single neurons in the orbitofrontal cortex enables such cells to show olfactory responses that reflect the taste association of the odor. PMID- 8727405 TI - Hunger and satiety modify the responses of olfactory and visual neurons in the primate orbitofrontal cortex. AB - 1. The primate orbitofrontal cortex is the site of convergence of information from primary taste and primary olfactory cortical regions. In addition, it receives projections from temporal lobe visual areas concerned with the representation of objects such as foods. Previous work has shown that the responses of gustatory neurons in the secondary taste area within the orbitofrontal cortex are modulated by hunger and satiety, in that they stop responding to the taste of a food on which an animal has been fed to behavioral satiation, yet may continue to respond to the taste of other foods. 2. This study demonstrates a similar modulation of the responses of olfactory and visual orbitofrontal cortex neurons after feeding to satiety. Seven of nine olfactory neurons that were responsive to the odors of foods, such as blackcurrant juice, were found to decrease their responses to the odor of the satiating food in a selective and statistically significant manner. 3. It also was found for eight of nine neurons that had selective responses to the sight of food, that they demonstrated a sensory-specific reduction in their visual responses to foods after satiation. 4. The responses of orbitofrontal cortex neurons selective for foods in more than one modality also were analyzed before and after feeding to satiation. Satiety often affected the responses of these multimodal neurons across all modalities, but a sensory-specific effect was not always demonstrable for both modalities. 5. These findings show that the olfactory and visual representations of food, as well as the taste representation of food, in the primate orbitofrontal cortex are modulated by hunger. Usually a component related to sensory-specific satiety can be demonstrated. The findings link at least part of the processing of olfactory and visual information in this brain region to the control of feeding-related behavior. PMID- 8727406 TI - Shared calcium signaling pathways in the induction of long-term potentiation and synaptic disinhibition in CA1 pyramidal cell dendrites. AB - 1. Calcium signaling pathways were examined in the induction of long-term synaptic disinhibition following tetanization. Effects of tetanization on gamma aminobutyric acid-A (GABAA receptor-mediated inhibitory responses were measured and compared with excitatory responses under experimental conditions previously used for examining induction mechanisms of N-methyl-D-aspartate (NMDA)-dependent long-term potentiation (LTP). Intracellular recordings were performed in current clamp and discontinuous single-electrode voltage-clamp (dSEVC) modes in CA1 pyramidal cell apical dendrites in hippocampal slices of adult guinea pigs with the use of sharp electrodes. Test pulses and tetanic stimuli were applied to the Schaffer collateral fibers in stratum radiatum. 2. Under standard control conditions [3 M K Ac in the recording pipette and artificial cerebrospinal fluid as extracellular solution], tetanization-induced sustained increases of excitatory responses were accompanied by marked decreases of parameters of GABAA mediated synaptic inhibition: at 40 min after tetanization [posttetanus 40 (PT 40)], orthodromically evoked excitatory postsynaptic potential (EPSP) peak amplitudes were on average 195 +/- 15% (mean +/- SE) and excitatory postsynaptic currents (IPSPs) were 166 +/- 10% of pretetanus controls. Peak amplitudes of orthodromically evoked inhibitory postsynaptic potentials (IPSPs) were 30 +/- 5% and inhibitory postsynaptic currents (IPSCs) were 21 +/- 4% at PT 40. Synaptic GABAA conductances (measured as chord conductances) were reduced to 22 +/- 4% at PT 40. Iontophoretic GABAA responses measured as conductance changes were 28 +/- 4% of pretetanus controls at PT 40. 3. A role of NMDA receptors in induction of long-term synaptic disinhibition was tested by preventing NMDA receptor activation 1) by pharmacological means and 2) by holding the membrane clamped at 80 mV (in dSEVC) during tetanization. In the presence of the NMDA-receptor antagonist D-2-amino5-phosphonopentanoic acid (D-AP5) 10-40 microM), orthodromically evoked EPSP amplitudes were 107 +/- 9%, EPSCs were 104 +/- 6%, GABAA-mediated IPSPs were 88 +/- 8%, IPSCs were 97 +/- 8%, synaptic GABAA conductances were 84 +/- 9%, and iontophoretic GABAA conductances were 102 +/- 13% at PT 40. In recordings in which the dendritic membrane potential was clamped at -80 mV during tetanization, orthodromically evoked peak amplitudes of EPSPs were 105 +/- 11%, EPSCs were 102 +/- 8, IPSPs were 103 +/- 4%, IPSCs were 102 +/- 5%, GABAA chord conductances were 101 +/- 9%, and iontophoretically evoked GABAA conductances were 105 +/- 5% at PT 40. 4. In recordings in which the intracellular pipette was preloaded with the Ca2+ chelator 1,2-bis(2 aminophenoxy) ethane-N,N,N'N"-tetraacetic acid (BAPTA) (5mM), long-term changes of synaptic transmission (increases of excitation, decreases of synaptic inhibition) were prevented. At PT 40, EPSP peak amplitudes were 93 +/- 7%, EPSCs were 115 +/- 6%, IPSPs were 115 +/- 9%, IPSCs were 117 +/- 8%, and synaptic GABAA conductances were 108 +/- 17%. Iontophoretic conductances at PT 40 were 109 +/- 9% over pretetanus controls when recorded with BAPTA-containing electrodes. 5. In recordings in which the intracellular pipette was preloaded with cypermethrin, a potent and selective inhibitor of phosphatase 2B, respective long-term changes of synaptic transmission (increases of excitation, decreases of synaptic inhibition) were prevented. At PT 40, EPSP peak amplitudes were 98 +/- 6%, EPSCs were 105 +/- 10%, IPSPs were 99 +/- 5%, IPSCs were 104 +/- 7%, synaptic GABAA conductances were 97 +/- 6% and iontophoretic GABAA conductances were 113 +/- 18% over pretetanus controls in cypermethrin-containing recordings. 6. In conclusion, the data presented demonstrate shared cellular pathways in the induction of both LTP and long-term synaptic disinhibition in apical dendrites of CA1 pyramidal cells after tetanization of the Schaffer collaterals. PMID- 8727407 TI - Impulse initiation in the mammalian muscle spindle during combined fusimotor stimulation and succinyl choline infusion. AB - 1. This is a report of observations on the responses of the primary and secondary endings of soleus muscle spindles of the anesthetized cat to the combined effects of the depolarizing neuromuscular blocker succinyl choline (SCh), given intravenously, and fusimotor stimulation. The findings were interpreted in terms of a dual pacemaker model for activity generated in the bag1 intrafusal fiber interacting with activity coming from bag2 and chain fibers. 2. In preliminary experiments it was found, using whole ventral root stimulation at fusimotor strength, that spindle responses to fusimotor stimulation were not blocked by SCh, whereas extrafusal junctions blocked rapidly. In the presence of SCh, fusimotor responses of spindle secondary endings were, on average, slightly larger than their control values before SCh was given, whereas fusimotor responses of primary endings were slightly smaller. 3. A study of the responses of spindle primary endings to stimulation of single dynamic (gamma D) and static (gamma S) axons in the presence of SCh revealed a fundamental difference in behavior. None of the responses to stimulation of gamma D axons (9 gamma D axons with 8 primary endings) showed significant summation with the responses to SCh. By contrast, the 20 gamma S axons studied showed varying degrees of summation with the responses to SCh. The responses of secondary endings to gamma S stimulation in the presence of SCh resembled those of primary endings and gamma S stimulation. 4. To explain these differences it is proposed that the primary ending has two separate sites of impulse initiation, one close to terminals on the bag1 intrafusal fiber (innervated by gamma D axons) and a second close to terminals on the bag2 and chain fibers (innervated by gamma S axons). It is proposed that the maintained increase in spindle firing observed during SCh infusion is the result of a bag2 contracture. The response to gamma S stimulation, contracting bag2 and chain fibers, adds to the SCh response. The degree of summation varies depending on whether the gamma S activates bag2 fibers, chain fibers, or both. The bag1 contracture, together with the effects of gamma D stimulation, acts through a separate pacemaker and therefore does not sum with the steady increase in spindle firing in the presence of SCh. There may be pacemaker switching between the bag1 generator and the bag2 and chain generator. 5. If the model is representative of most spindles containing the three kinds of intrafusal fibers, and the contractions of bag2 and chain fibers generate activity through a common impulse generator, then this bears on the question of the functional independence of the bag2 and chain fiber systems. PMID- 8727408 TI - Time course of experience-dependent synaptic potentiation and depression in barrel cortex of adolescent rats. AB - 1. Plasticity could be induced in (S1) barrel cortex of adolescent rats by reducing the complement of vibrissae on one side of the muzzle to a single whisker for a period of 7, 20, or 60 days. The effect of deprivation was assessed by quantitatively by measuring cortical responses to stimulation of the spared and regrown deprived vibrissae. Vibrissa responses were evoked using a standard stimulus generated by an electromechanical stimulator and measured using poststimulus time histogram analysis. 2. Cells located in layers II/III were found to be plastic beyond postnatal day 28 (P28), whereas cells located in layer IV were not. The vibrissa dominance distribution was shifted significantly toward the spared vibrissa after 7, 20, and 60 days of deprivation for cells located in layers II/III of barrel columns surrounding the D1 column (P < 0.0001, 2-factor analysis of variance). The vibrissa dominance distribution did not shift significantly for cells located in layer IV of surrounding barrels for any of the durations of deprivation tested (P > 0.1). After 7 days of deprivation, 37% of the cells located in layers II/III of the columns deprived vibrissae showed greater responses to the spared vibrissae than to their deprived principal vibrissa, compared with 11% in normally reared adolescent animals and 3% in adults. The percentage of cells dominated by the spared vibrissa was 65% after 20 days of deprivation and 43% after 60 days. 3. For cells located in layers II/III, short-term deprivation (7 days) caused a decrease in the absolute magnitude of response to stimulation of the deprived vibrissa (reduction to approximately 28% of control levels). However, no change could be detected in the spared (D1) vibrissa input to the same deprived columns. Therefore the increase in D1 dominance registered in the deprived columns was mainly due to a decrease in principal vibrissa response and no change in the spared D1 vibrissa response. 4. The first increase in spared vibrissa response was seen after 20 days of deprivation. The response magnitude cells located in layers II/III increased to 70% above control levels. Responses to deprived vibrissa stimulation were depressed at 20 days of deprivation (reduction to 35% of control), implying that the vibrissa dominance shift at 20 days was due to both an increase in spared vibrissa response and a decrease in deprived vibrissa response. 5. The spared vibrissa response was increased after 60 days of deprivation (110% above control) in both near and far halves of the barrel columns surrounding D1. On average, the deprived vibrissa response was depressed at 60 days (84% of control), although less than at 20 or 7 days, because of recovery of responsiveness in the far half of the deprived barrel column. Layer II/III cells located in the half of the barrel column farthest from the spared D1 barrel column showed normal levels of deprived vibrissa input, whereas cells located in the half of the barrel column closest to the spared D1 barrel column still exhibited depressed levels of deprived vibrissa input (54% of control). 6. Control experiments suggested that depression of the deprived vibrissa response could not be explained by nonspecific effects. Depression was not a function of animal's age, because normally reared P28 and adult animals showed similar principal vibrissa response levels. It was not a result of nonspecific depression of cortical responses. because the decreased response only occurred in cells of deprived barrel columns and not spared barrel columns (recorded in the same animals). Depression was not due to altered vibrissa mechanics, because the spared vibrissa response was similarly depressed in animals in which the vibrissae had been trimmed rather than removed. Finally, depression was input specific at 7 and 20 days, because only the deprived vibrissa responses were depressed, whereas spared vibrissa responses were either at control levels or at elevated levels for the same cells. 7. PMID- 8727409 TI - Neural systems for tactual memories. AB - 1. The aim of this study was to investigate the neural systems involved in the memory processing of experiences through touch. 2. Regional cerebral blood flow was measured with positron emission tomography by means of the water bolus H2(15)O methodology in human subjects as they performed tasks involving different levels of tactual memory. In one of the experimental tasks, the subjects had to palpate nonsense shapes to match each one to a previously learned set, thus requiring constant reference to long-term memory. The other experimental task involved judgements of the recent recurrence of shapes during the scanning period. A set of three control tasks was used to control for the type of exploratory movements and sensory processing inherent in the two experimental tasks. 3. Comparisons of the distribution of activity between the experimental and the control tasks were carried out by means of the subtraction method. In relation to the control conditions, the two experimental tasks requiring memory resulted in significant changes within the posteroventral insula and the central opercular region. In addition, the task requiring recall from long-term memory yielded changes in the perirhinal cortex. 4. The above findings demonstrated that a ventrally directed parietoinsular pathway, leading to the posteroventral insula and the perirhinal cortex, constitutes a system by which long-lasting representations of tactual experiences are formed. It is proposed that the posteroventral insula is involved in tactual feature analysis, by analogy with the similar role of the inferotemporal cortex in vision, whereas the perirhinal cortex is further involved in the integration of these features into long-lasting representations of somatosensory experiences. PMID- 8727410 TI - Diffusion, not uptake, limits glycine concentration in the synaptic cleft. AB - 1. The question of whether active uptake limits the duration of action of the inhibitory transmitter glycine has been addressed in vivo at inhibitory synapses on the goldfish Mauthner (M) cell. The kinetics of inhibitory postsynaptic potentials and inhibitory postsynaptic currents (IPSCs) evoked antidromically and by eighth-nerve stimulation were recorded in control and in conditions expected to block glycine uptake or slow its diffusion. Theoretical considerations, based on simulated quantal currents, predicted that if diffusion was slow, rapid uptake of glycine would be required and its block would prolong the synaptic responses. 2. Temperature coefficient values for IPSC decay time constants (tau S) are in the range of 2.0 for temperatures between 15 and 23 degrees C, suggesting that diffusion is not the rate-limiting step. 3. Li+, Ch+, or N-methyl-D-glucamine were substituted for 80% of the Na+ in the extracellular fluid to analyze the effects of blocking the Na(+)-dependent glycine uptake. These procedures enhanced the maximum inhibitory shunt produced by glycine iontophoresis, leading to the suggestion that uptake may buffer the concentration of the transmitter in the cleft. In contrast, the Na+ substitutes had no effect on the tau of the recurrent collateral IPSC, which involves synchronous activation of a pool of interneurons and has a monoexponential decay (tau approximately 10-11 ms). 4. The decay phase of the disynaptic IPSCs produced by stimulating the contralateral eighth nerve has fast and slow components, with a prolonged tail lasting up to 100 ms, particularly in the case of repetitive nerve stimulation. The tail is inhibitory, as revealed by its shunt of the antidromic action potential, and it is at least partially Cl- dependent. However, it can be accelerated by superfusion with the glutamate receptor antagonists 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) and DL 2-amino-5-phosphonopentanoic acid (APV). In the presence of these blockers, the IPSC decay remains biexponential (tau fast = 5.2 and 5.9 ms, tau slow = 94 and 130 ms for single and burst stimuli, respectively). Blocking uptake in this condition did not modify tau fast or tau slow. 5. We conclude that an active uptake mechanism does not shape glycinergic IPSCs, including the longer-lasting components that might include a contribution due to persistence of the transmitter. Rather, diffusion alone is sufficient to remove glycine at a rate faster than channel unbinding. 6. To test whether glycine might diffuse to adjacent excitatory synapses and enhance activation of N-methyl-D-aspartate receptors, CNQX and APV were applied locally, by pressure, to the M cell soma, but they had no effect on the prolonged decay of eighth-nerve-evoked responses. Thus the effects of the antagonists when added to the superfusate are exerted at the network level. PMID- 8727411 TI - Somatosensory cortical efferent neurons of the awake rabbit: latencies to activation via supra--and subthreshold receptive fields. AB - 1. Latencies to peripheral sensory stimulation were examined in four classes of antidromically identified efferent neurons in the primary somatosensory cortex (S1) of awake rabbits. Both suprathreshold responses (action potentials) and subthreshold responses were examined. Subthreshold responses were examined by monitoring the thresholds of efferent neurons to juxtasomal current pulses (JSCPs) delivered through the recording microelectrode (usually 1-3 microA). Through the use of this method, excitatory postsynaptic potentials (EPSPs) and inhibitory postsynaptic potentials (IPSPs) were manifested as decreases and increases in threshold, respectively. Efferent populations examined included callosal (CC) neurons, ipsilateral corticocortical (C-IC) neurons, and descending corticofugal neurons of layer 5 (CF-5) and layer 6 (CF-6). Very brief air puffs (rise and fall times 0.6 ms) were delivered to the receptor periphery via a high speed solenoid valve. 2. Whereas all CF-5 neurons had demonstrable suprathreshold excitatory and/or inhibitory responses to peripheral stimulation, most CC, C-IC, and CF-6 neurons did not. CC and CF-6 neurons that yielded no suprathreshold response to the stimulus had lower axonal conduction velocities than those that did respond (P < 0.0001 in both cases). However, subthreshold receptive fields could be demonstrated in many of the otherwise unresponsive CC (81%), C-IC (88%), and CF-6 (43%) neurons. The subthreshold responses usually consisted of an initial excitatory component (a decrease in the threshold to the JSCP) and a subsequent long-duration (> 80 ms) inhibitory component. A few neurons (1 CC, 1 C IC, and 5 CF-6) showed an initial short latency inhibitory response in the absence of any excitatory component. 3. Some CC and C-IC neurons yielded supra- and/or subthreshold responses to peripheral stimulation at latencies of 6.1-7 ms. All such neurons were found at intermediate cortical depths (thought to correspond to deep layer 2-3 through layer 5). It is argued that such latencies are indicative of monosynaptic activation via thalamic afferents. Very superficial CC and C-IC neurons, and all CF-6 neurons responded to latencies of > 7 ms. All CF-5 neurons responded to latencies of > 8 ms, although many were found at the same depth as the deeper CC and C-IC neurons that responded at monosynaptic latencies. These results indicate that cortical cell type as well as laminar position are important factors that determine the sequence of intracortical neuronal activation after peripheral sensory stimulation. PMID- 8727412 TI - Coordinating movement at two joints: a principle of linear covariance. AB - 1. Six subjects performed fast, "single-joint" flexions at either the elbow or shoulder over three angular distances in a sagittal plane. Movement endpoints were located to require flexion of only a single, "focal" joint, without any external, mechanical constraint on the other, "nonfocal" joint. Three subjects performed another series of movements between two targets while moving along different paths and in which both joints were flexed. 2. We compared the torque patterns that were produced at the two joints. For single-joint movements, they were both biphasic pulses that accelerated and then decelerated the limb. 3. The torque at the nonfocal joint of a single joint movement was very close to linearly proportional to that at the focal joint throughout the movement. Elbow and shoulder torques differed by a linear scaling constant and went through extrema and zero crossings almost simultaneously. 4. In contrast, during movements in which subjects were explicitly instructed to use a hand path they would not naturally, use the linear interjoint torque scaling rule did not apply. This demonstrated that when we wish to move along a path between two targets that is not produced by linear torque covariation, we are able to modify that rule at will. 5. We speculate that linear, dynamic covariation of the torque patterns across two joints may be an important principle for reducing the number of degrees of freedom that the nervous system must independently control in performing unconstrained limb movements over naturally chosen paths. PMID- 8727413 TI - Detection of diffuse breast cancers with a new sonographic method. AB - The presentation of diffuse cancers detected with a new sonographic method is described. It uses compression of the breast in positions which are used in mammography and a reflecting metallic plate to look for the presence of breast lesions. Changes in reflectivity and velocity are imaged on this reference structure and can be compared with appearance of the reference structure behind normal tissue. Detectability and sonographic appearance are reported in 13 diffuse malignancies and one diffuse benign lesion that had been found among 89 malignant and 83 benign lesions in 151 preoperative patients. Ten breast cancer patients who had been treated with breast-conserving surgery and radiation therapy were also examined sonographically as a model for diffuse changes of the breast parenchyma. PMID- 8727414 TI - Comparison of transrectal ultrasound prostatic volume estimation with magnetic resonance imaging volume estimation and surgical specimen weight in patients with benign prostatic hyperplasia. AB - There are relatively few reports in the literature comparing transrectal ultrasound (TRUS) to magnetic resonance imaging (MRI) in estimating the volume of the prostate in the management of benign prostatic hyperplasia (BPH). In this study, we compared volumes determined by TRUS with MRI and TRUS-estimated weights with surgical specimen weights. The main findings of this study were (a) TRUS and MRI measurement of prostate volumes are quite similar; and (b) TRUS underestimates (by 10%) the prostatic weight as determined from the surgical specimens. A better volume estimate can thus be obtained by multiplying the TRUS measurement by a factor of 1.10. We conclude that because TRUS is cheap, user friendly, noninvasive, and equally as accurate as MRI, it should be the preferred modality in the follow-up of BPH patients. PMID- 8727415 TI - Follow-up of small, postmenopausal ovarian cysts using vaginal ultrasound and CA 125 antigen. AB - To determine the natural history of small, simple ovarian cysts in postmenopausal women, 51 postmenopausal patients with small (< 5 cm), smooth, aseptate, hypoechogenic ovarian cysts, without ascites, were followed by vaginal ultrasound examinations every 3 months for an average period of 2.5 years. In 34, CA-125 antigen was measured and found to be within normal limits. None of the cysts showed changes in texture, nor did ascites appear. The CA-125 antigen serum levels remained low. The mean size of the cysts decreased with time. There was no statistically significant correlation between the initial size of the cyst, its tendency to grow or shrink, and the absolute CA-125 serum level. Our findings support the option of a conservative follow-up by repeated ultrasonic and CA-125 antigen examinations of small, simple cysts in postmenopausal women. PMID- 8727416 TI - Ultrasonographic "endometrial three-layer" pattern: a unique finding in ectopic pregnancy. AB - Although diagnostic ultrasonography is playing an increasing role in the investigation of the patient with suspected ectopic pregnancy (EP), it has significant limitations in the very early stages of pregnancy. By sonographically exploring the intrauterine echoes in 45 cases of documented EP, we demonstrated a unique pattern in 28 cases (62.2%). A well-defined spheric structure forming an endometrial three-layer (ETL) pattern was seen, probably formed by a midline echo between the two adjacent edematous proliferative layers of the endometrium, the latter resembling the late proliferative endometrium. In 17 patients with a proved EP (37.8%), the ETL pattern was not demonstrated. However, the ETL was not demonstrated in all 40 cases of early intrauterine pregnancy and all 50 cases of miscarriage. These findings suggest a 100% specificity and a sensitivity of 62.2% for the ETL pattern in the diagnosis of EP. PMID- 8727417 TI - Flow velocity waveforms of the ductus arteriosus in appropriate and small-for gestational-age fetuses. AB - OBJECTIVES: To assess ductus arteriosus velocity waveforms in the appropriate and small-for-gestational-age (SGA) fetuses. STUDY DESIGN: Ductus arteriosus flow velocity waveforms were obtained in 14 normal fetuses in a longitudinal study and in 139 normal fetuses in a cross-sectional study with pulsed and continuous Doppler ultrasonography. Ductus velocity waveforms were also determined in 25 SGA fetuses whose gestational age ranged between 20 and 34 weeks (mean +/- SD: 30 +/- 4.1 weeks). RESULTS: Mathematical modeling demonstrated that the peak systolic velocity, lowest diastolic velocity, and mean velocity of the ductus arteriosus increased with advancing gestation in both the longitudinal and cross-sectional study, but considerable individual variability was seen in the longitudinal study. The pulsatility index did not change significantly. The ductal parameters were in the normal range in all the SGA fetuses. CONCLUSIONS: The blood velocity of the ductus arteriosus increase with advancing gestation. Our results suggest that Doppler ultrasonographic study of the fetal ductus arteriosus, unlike Doppler study of the umbilical artery or middle cerebral artery, is not useful as an indicator of adverse fetal outcome. PMID- 8727418 TI - Testicular microlithiasis: a benign condition with a malignant association. AB - Testicular microlithiasis (TM), which usually affects both testes, is diagnosed primarily by ultrasound. TM has been found to be associated with benign conditions but has also been reported in association with testicular cancer. Echographically, TM is manifested by diffuse, punctate, non-shadowing, hyperechoic foci within the testicular parenchyma. To date, more than 80 cases of TM have been documented sonographically or pathologically; 25 of them have demonstrated an association with pathologically proven testicular cancer (p < 0.05). Thirty-two cases were associated with infertility/subfertility (p < 0.05). We believe the finding of TM on ultrasound should heighten the radiologist's suspicion of testicular malignancy. If no malignancy is identified on initial evaluation including CT of the chest and abdomen, close clinical follow-up with periodic (every 6 to 12 months) scrotal ultrasound examination is probably indicated. PMID- 8727419 TI - Giant hyperplastic polyp of the gallbladder: a case report. PMID- 8727420 TI - Synovial osteochondromatosis of the distal radioulnar joint: ultrasound diagnosis. PMID- 8727421 TI - Transvaginal ultrasonographic findings in vesico-uterine fistula. PMID- 8727422 TI - Ultrasonographic diagnosis of small intestinal phytobezoar. PMID- 8727423 TI - Usefulness of ultrasonography and magnetic resonance imaging for prenatal diagnosis of fetal teratoma of the neck. PMID- 8727424 TI - Endosonography and ovarian endometriosis. PMID- 8727425 TI - Prodrugs revisited: the "ad hoc" approach as a complement to ligand design. PMID- 8727426 TI - Design of drugs involving the concepts and theories of drug metabolism and pharmacokinetics. AB - Drug metabolism input to the discovery process had historically been on an empirical case-by-case basis, since, detailed descriptors of the effect on pharmacokinetics of a change in structure or physicochemical property were not available. Considerable advances have been made in recent years, such that basic rules can be applied to predict the behavior of a compound in man based on physicochemistry and structure. This is particularly true in the areas of absorption, distribution, and clearance. In particular, knowledge of the reactions catalyzed by the enzymes of drug metabolism, including the cytochrome P450 super family, can be used in the design of new chemical entities, together with the usual pharmacological-derived SAR. The combination of both pharmacokinetics and pharmacodynamics at the discovery stage leads to drugs with optimum performance characteristics. Such drugs are easier to develop, representing a huge saving in resources. Moreover, the marketed compound is much more likely to find high clinical utilization. This review uses dofetilide, fluconazole, and amlodipine to highlight the multifaceted consequences of changing chemical structure, in terms of drug disposition, and reinforces these principles with examples from the literature. PMID- 8727427 TI - Quantitative structure--activity relationship (QSAR) studies of mutagens and carcinogens. PMID- 8727428 TI - Topical iontophoretic drug delivery: pathways, principles, factors, and skin irritation. PMID- 8727429 TI - Permissive hypercapnia in ARDS: just do it? PMID- 8727430 TI - Effects of rapid permissive hypercapnia on hemodynamics, gas exchange, and oxygen transport and consumption during mechanical ventilation for the acute respiratory distress syndrome. AB - OBJECTIVE: To measure the effects of rapid permissive hypercapnia on hemodynamics and gas exchange in patients with acute respiratory distress syndrome (ARDS). DESIGN: Prospective study. SETTING: 18-bed, medical intensive care unit, university hospital. PATIENTS: 11 mechanically ventilated ARDS patients. INTERVENTION: Patients were sedated and ventilated in the controlled mode. Hypercapnia was induced over a 30-60 min period by decreasing tidal volume until pH decreased to 7.2 and/or P50 increased by 7.5 mmHg. Settings were then maintained for 2 h. RESULTS: Minute ventilation was reduced from 13.5 +/- 6.1 to 8.2 +/- 4.1 l/min (mean +/- SD), PaCO2 increased (40.3 +/- 6.6 to 59.3 +/- 7.2 mmHg), pH decreased (7.40 +/- 0.05 to 7.26 +/- 0.05), and P50 increased (26.3 +/- 2.02 to 31.1 +/- 2.2 mmHg) (p < 0.05). Systemic vascular resistance decreased (865 +/- 454 to 648 +/- 265 dyne.s.cm-5, and cardiac index (CI) increased (4 +/- 2.4 to 4.7 +/- 2.4 l/min/m2) (p < 0.05). Mean systemic arterial pressure was unchanged. Pulmonary vascular resistance was unmodified, and mean pulmonary artery pressure (MPAP) increased (29 +/- 5 to 32 +/- 6 mmHg, p < 0.05). PaO2 remained unchanged, while saturation decreased (93 +/- 3 to 90 +/- 3%, p < 0.05), requiring an increase in FIO2 from 0.56 to 0.64 in order to maintain an SaO2 > 90%. PvO2 increased (36.5 +/- 5.7 to 43.2 +/- 6.1 mmHg, p < 0.05), while saturation was unmodified. The arteriovenous O2 content difference was unaltered. Oxygen transport (DO2) increased (545 +/- 240 to 621 +/- 274 ml/min/m2, p < 0.05), while the O2 consumption and extraction ratio did not change significantly. Venous admixture (Qva/Qt) increased (26.3 +/- 12.3 to 32.8 +/- 13.2, p < 0.05). CONCLUSIONS: These data indicate that acute hypercapnia increases DO2 and O2 off-loading capacity in ARDS patients with normal plasma lactate, without increasing O2 extraction. Whether this would be beneficial in patients with elevated lactate levels, indicating tissue hypoxia, remains to be determined. Furthermore, even though hypercapnia was well tolerated, the increase in Qva/Qt, CI, and MPAP should prompt caution in patients with severe hypoxemia, as well as in those with depressed cardiac function and/or severe pulmonary hypertension. PMID- 8727431 TI - Physiological effects of reduced tidal volume at constant minute ventilation and inspiratory flow rate in acute respiratory distress syndrome. AB - OBJECTIVE: To assess the effect of changes in tidal volume (VT) with a constant inspiratory flow and minute ventilation (VE) on gas exchange and oxygen transport in acute respiratory distress syndrome (ARDS). DESIGN: A crossover study of three VT in two study groups, using patients as their own controls. SETTING: A medical surgical intensive care unit in a tertiary care center. PATIENTS: Eight patients with ARDS and seven postoperative cardiac surgery patients with uncomplicated recoveries were studied during volume-controlled mechanical ventilation. INTERVENTIONS: During controlled mechanical ventilation, patients were first ventilated with a VT of 9-11 ml/kg. VT was then increased to 12-14 ml/kg (+ 25%) for 30 min and subsequently decreased to 6-8 ml/kg (- 25%) for 30 min by adjusting the respiratory rate (RR) while the inspiratory flow rate, VE, and inspiratory duty cycle (TL/T(TOT)) were kept constant. At the end, patients were ventilated with the baseline settings for another 30 min. MEASUREMENTS AND RESULTS: VE, carbon dioxide production (VCO2) and oxygen consumption (VO2) were measured continuously with a gas exchange monitor, and cardiac output and arterial and mixed venous blood samples were taken at the end of each 30-min period to assess CO2 removal and oxygen transport. Alveolar minute ventilation (VA) and the deadspace to tidal volume ratio (VD/VT) were calculated from the Bohr equation. Despite large changes in VT, arterial oxygenation (PaO2) and oxygen transport (DO2) were unchanged throughout the study. When VT was increased, physiological VD increased from 448 +/- 34 ml to 559 +/- 46 ml (mean +/- SE) in ARDS (P < 0.001) and from 281 +/- 22 ml to 357 +/- 35 ml in CABG (P < 0.05). With the small VT, VD decreased to 357 +/- 22 ml in ARDS (P < 0.01), and to 234 +/- 24 ml in CABG (P < 0.05). In ARDS, VD/VT decreased from 0.57 +/- 0.03 to 0.55 +/- 0.03 (P < 0.05) with the large VT, and increased to 0.60 +/- 0.03 (P < 0.01), when VT was reduced. In CABG, VD/VT did not change significantly. ARDS patients had a higher PaCO2 than cardiac patients (P < 0.001), and only minor changes in PaCO2 were observed (for ARDS and CABG respectively, baseline 5.9 +/- 0.3 kPa and 4.1 +/- 0.1 kPa, large VT 5.7 +/- 0.3 kPa and 4.1 +/- 0.2 kPa, small VT 6.2 +/- 0.3 kPa and 4.2 +/- 0.2 kPa; P < 0.05). CONCLUSIONS: Tidal volumes can be reduced to 6-8 ml/kg in ARDS patients without compromising oxygen transport, while adequate CO2 elimination can be maintained. PMID- 8727432 TI - The automatic selection of ventilation parameters during the initial phase of mechanical ventilation. AB - OBJECTIVE: To test a method that allows automatic set-up of the ventilator controls at the onset of ventilation. DESIGN: Prospective randomized crossover study. SETTING: ICUs in one adult and one children's hospital in Switzerland. PATIENTS: Thirty intubated stable, critically ill patients (20 adults and 10 children). INTERVENTIONS: The patients were ventilated during two 20-min periods using a modified Hamilton AMADEUS ventilator. During the control period the ventilator settings were chosen immediately prior to the study. During the other period individual settings were automatically determined by the ventilatior (AutoInit). MEASUREMENTS AND RESULTS: Pressure, flow, and instantaneous CO2 concentration were measured at the airway opening. From these measurements, series dead space (V(DS)), expiratory time constant (RC), tidal volume (VT, total respiratory frequency (f(tot), minute ventilation (MV), and maximal and mean airway pressure (Paw, max and Paw, mean) were calculated. Arterial blood gases were analyzed at the end of each period. Paw, max was significantly less with the AutoInit ventilator settings while f(tot) was significantly greater (P < 0.05). The other values were not statistically significant. CONCLUSIONS: The AutoInit ventilator settings, which were automatically derived, were acceptable for all patients for a period of 20 min and were not found to be inferior to the control ventilator settings. This makes the AutoInit method potentially useful as an automatic start-up procedure for mechanical ventilation. PMID- 8727433 TI - Computed tomography vs clinical and multidisciplinary procedures for early evaluation of severe abdomen and chest trauma--a cost analysis approach. AB - OBJECTIVE: To compare contrast computed tomography (CT) for evaluating abdominal and vascular chest injuries after emergency room resuscitation with multidisciplinary management based on bedside procedure (BP), e.g., peritoneal lavage, abdomen ultrasonography urography and, if indicated, CT and/or aortography or transesophageal echocardiography. DESIGN: Randomized study. SETTING: Emergency, critical care and radiology departments in a trauma center. PATIENTS: The study was performed in 103 severe blunt trauma patients with a revised trauma index < 8, admitted over a 16 month period and divided into group (G1, n = 52, CT management) and group 2 (G2, n = 51, BP management). INTERVENTIONS: A relative direct cost scale used in our trauma center was applied, and cost units (U) were assigned to each diagnostic test for cost minimization analysis (abdomen ultrasonograph = 7.5 U, peritoneal lavage = 8 U, urography = 9 U, computed tomography = 9 U, transesophageal echocardiography = 13.5 U, and aortography = 15 U). One unit is approximately equivalent to $43.7. RESULTS: Injury severity score (ISS) was 31.7 +/- 15.4 in G1 and 33.8 +/- 18.3 in G2. Sensitivity for CT was 90.4% (G1) vs 72.5% for BP (G2) in abdomen (P < 0.01) and 60% in chest for evaluating mediastinal hematoma etiology (G1). As Table 2 shows, G1 needed 59 tests for evaluating injuries (1.1 +/- 0.3 tests patient) while G2 required 81 tests (1.68 +/- 0.8 tests/patient) (P < 0.01). The total relative cost was 538 U for G1, 7.04 +/- 2.2 U cost/injury and 10.3 +/- 3.3 U/evaluation of trauma vs 698 U for G2, 9.84 +/- 5.03 U cost/injury and 13.68 +/- 8.5 U/evaluation (P < 0.05). CONCLUSIONS: This cost-minimization study suggests that CT is a more cost-effective method for the post-emergency room resuscitation evaluation of severe abdominal blunt trauma than the multidisciplinary BP. Chest CT is a screening method for mediastinal hematoma but not for etiology. PMID- 8727434 TI - Renal effects of low-dose dopamine in patients with sepsis syndrome or septic shock treated with catecholamines. AB - OBJECTIVE: To evaluate the renal effects of low-dose dopamine in patients with sepsis syndrome or septic shock treated with catecholamines. DESIGN: Prospective, clinical study using sequential periods. SETTING: A 12-bed surgical intensive care unit in a university hospital. PATIENTS: 14 patients with sepsis syndrome and 15 patients with septic shock treated with exogenous catecholamines were studied. They had no diuretic treatment. INTERVENTION: Two periods of 2 h each with and without 2 micrograms.kg-1.min-1 of dopamine infusion. Hemodynamic and renal data were obtained at the end of each period. Measurements were repeated after 48 h of dopamine infusion in patients with sepsis syndrome. All data were evaluated by the Wilcoxon rank test. MEASUREMENTS AND RESULTS: In patients with sepsis syndrome, diuresis and creatinine clearance increased significantly by 100% and 60%, respectively, during low-dose dopamine infusion without any change in systemic hemodynamics. The renal response to dopamine decreased significantly after 48 h of dopamine infusion (P < 0.01). In patients with septic shock treated with catecholamines, no variation of either systemic hemodynamics or renal function was noted during low-dose dopamine infusion. CONCLUSION: The renal effects of low-dose dopamine in patients with sepsis syndrome decrease with time. No renal effect of low-dose dopamine was observed in patients with septic shock treated with catecholamines. These findings suggest a desensitization of renal dopaminergic receptors. PMID- 8727435 TI - Assessment of gastric acidity in intensive care patients: intermittent pH registration cannot replace continuous pH monitoring. AB - OBJECTIVE: To test the accuracy of colour-scaled indicator papers to measure pH values and to study the correlation between this method of measuring gastric juice pH once daily and 24-h continuous intragastric pH monitoring in intensive care patients. DESIGN: The accuracy of indicator papers was tested in the laboratory using colourless solutions and aspirated gastric juice and was then verified with a laboratory pH meter. Continuous intragastric pH monitoring was performed in mechanically ventilated ICU patients. Percentages of time with a pH value <3.0 and median pH values by 24-h continuous intragastric pH monitoring were compared to pH values measured once daily with indicator paper. SETTING: A mixed ICU. PATIENTS: A total of 150 measurements were taken by continuous pH monitoring in 91 mechanically ventilated ICU patients. MEASUREMENTS AND RESULTS: The correlation between the pH measured with the indicator paper and subsequently verified with a laboratory pH meter in colourless solutions was 0.96 [regression coefficient (RC) 0.98, 95% confidence interval (CI) 0.91-1.05]. Measured in gastric juice it was 0.95 (RC 0.95, 95% CI 0.88-1.01). The correlation between median pH values, determined with 24-h continuous intragastric pH monitoring, and values measured with indicator papers was 0.39 (RC 0.43, 95% CI 0.26-0.59). The mean difference in pH, as determined by the analysis of Bland and Altman], was 0.9 with a SD of 4.7. The correlation between the percentage of time with pH < 3.0, as obtained with continuous registration, and median gastric pH values (also obtained with continuous registration) was -0.94 (RC-0.06, 95% CI-0.06- -0.05); the correlation between the time and gastric pH values (measured with indicator paper) was-0.40 (RC-0.02, 95% CI-0.03- -0.02). CONCLUSION: The colour-scaled indicator paper is an accurate method of measuring pH values, but there is a poor correlation between gastric pH values measured once daily and a total measurement derived from 24-h continuous intragastric pH monitoring. Changes in intragastric pH values cannot be accurately studied when measuring acidity once daily. The influence of various treatment regimens on intragastric acidity in relation to the development of gastric colonization and nosocomial pneumonia should be investigated either with continuous intragastric monitoring or with frequent measurements in aspirated gastric juice. PMID- 8727436 TI - Elevated serum bleomycin-detectable iron concentrations in patients with sepsis syndrome. AB - OBJECTIVE: To determine serum bleomycin-detectable ?free' iron in patients with septic shock and to relate these findings to both outcome and a marker of free radical damage. DESIGN: A prospective observational study. SETTING: A nine-bed intensive care unit in a university teaching hospital. PATIENTS: Sixteen consecutive patients with septic shock, defined as: (1) Clinical evidence of acute infection; (2) hypo- or hyperthermia ( < 35.6 degrees or > 38.3 degrees C); (3) tachypnoea ( > 20 breaths/min or ventilated); (4) tachycardia ( > 90 beats min); (5) shock (systolic pressure < 90mmHg) or on inotropes. Fourteen patients also had secondary organ dysfunction. MEASUREMENTS AND RESULTS: Bleomycin detectable iron concentrations were elevated in all patients (37.2 +/- 11.0 mumols/l vs 5.1 +/- 3.3 mumols/l in healthy subjects, P < 0.0001), but there was no difference between patients who died and those who survived (39.2 +/- 9.3 and 36.2 +/- 12.3 mumols/l, respectively). Thiobarbituric acid reactive substances (an index of lipid peroxidation) were higher in those who died (3.33 +/- 2.29 mumols/l) than in the surviving patients (0.99 +/- 0.14 mumols/l, P < 0.01) or healthy subjects (0.92 +/- 0.39 mumols/l, P < 0.01). Free iron did not correlate with thiobarbituric acid-reactive substances. However, a significant correlation was found between lipid peroxidation and clinical severity (APACHE II) score (r = 0.54, P < 0.05). CONCLUSIONS: The present study provides evidence of lipid peroxidation in patients who die with septic shock. The data suggest that iron catalysed hydroxyl radical generation does not form an important contribution to this lipid peroxidation in patients with sepsis. PMID- 8727437 TI - Effects of atrial natriuretic peptide on acute renal impairment in patients with heart failure after cardiac surgery. AB - OBJECTIVE: To investigate the effects of IV infusion of atrial natriuretic peptide (human ANP 1-28) on renal function in patients with acute heart failure and renal impairment after cardiac surgery. DESIGN: Pharmocodynamic dose-effect investigation. SETTING: Cardiothoracic Intensive Care Unit of a university hospital. PATIENTS: Twelve patients (mean age 68 years, range 44-78 years) treated with inotropic drugs and an intra-aortic balloon pump (n = 8) were studied 1-3 days after cardiac surgery. Patients had acute renal impairment, defined as a rise in serum creatinine of more than 50% compared to preoperative values. Patients were receiving dopamine and furosemide infusion to increase urine flow. INTERVENTIONS: Baseline measurements of glomerular filtration rate (GFR) and renal blood flow (51Cr-EDTA and PAH clearance) were first performed during two 30-min periods. ANP was then administered for two consecutive 30-min periods (25 and 50 ng/kg per min), followed by two control periods. MEASUREMENTS AND MAIN RESULTS: Mean arterial pressure decreased by 6% at the highest ANP dose. Urine flow, GFR and RBF increased 62%, 43% and 38%, respectively, while renal vascular resistance decreased 30%. At this dose level, circulating ANP concentrations were on the average eight fold higher than preinfusion levels. CONCLUSIONS: ANP improved renal function and decreased elevated renal vascular resistance in patients with renal dysfunction after cardiac surgery. The improvement in renal blood flow and glomerular filtration rate may be of potential therapeutic value to prevent or treat exaggerated renal vasoconstriction in patients with acute renal impairment following cardiac surgery. PMID- 8727438 TI - The prognosis of oncologic patients in the pediatric intensive care unit. AB - OBJECTIVE: To evaluate the predicted mortality rate of oncologic patients in the PICU using the PRISM score and factors that might influence short-term outcomes. DESIGN: Retrospective study. SETTING: Pediatric ICU in a university hospital. PATIENTS AND METHODS: The medical charts of all oncologic patients admitted to the PICU during the period from January 1983 to December 1992 were reviewed. MAIN RESULTS: Over a period of 10 years, 51 oncologic patients were admitted on 57 occasions to the PICU. The mortality was 32%. This is significantly higher than the overall mortality in the PICU (8%). Comparison of observed and predicted mortality, derived from the PRISM score, using chi square goodness-of-fit tests showed a significantly higher observed mortality (x2(5) = 20.1, P < 0.01). Patients admitted for circulatory failure had the highest mortality (47%), followed by those with respiratory failure due to tachypnea/cyanosis (36%), central nervous system deterioration (27%), respiratory failure due to airway obstruction (25%), and metabolic disorders (20%). Of the 31 patients who needed mechanical ventilation, 17 died (55%), and when they needed inotropic support as well, the mortality increased to 69%. The mortality rose to 100% when the patient was admitted with a septic shock, necessitating mechanical ventilation and inotropic support. The median PRISM score was 5 in the survivor group and 18.5 in the non-survivor group; this difference was found to be significant using the Wilcoxon test (P = 0.01). However, some patients with high scores were found in the survivor group, as well as some with low scores in the non-survivor group. CONCLUSION: The decision to treat oncologic patients in a PICU remains difficult and has to be considered on an individual basis. However, oncologic patients do benefit from admission to the PICU. The PRISM score is not suitable for oncologic patients in the PICU, because it underestimates the observed mortality. Other factors like neutropenia, septic shock, the need for mechanical ventilation, and inotropic support should be taken into consideration. PMID- 8727439 TI - Accuracy of oscillometric blood pressure measurement in critically ill neonates with reference to the arterial pressure wave shape. AB - OBJECTIVE: To perform further evaluation of the oscillometric device for neonatal arterial blood pressure (ABP) measurement, using a catheter-manometer system (CMS) for accurate intraarterial measurement. We aimed to describe the influence of the radial artery wave shape on oscillometric ABP determination, as pressure wave-shape influences the relationships between systolic arterial pressure (SAP), diastolic arterial pressure (DAP) and mean arterial pressure (MAP) in the wave. These relationships are part of the algorithms contributing to the final ABP determination in the oscillometric device. DESIGN: Intra-patient comparison of two blood pressure measurement systems. SETTING: Neonatal intensive care unit. PATIENTS: In 51 critically ill newborn infants, ABP was determined oscillometrically in the brachial artery and, simultaneously, invasively in the radial artery using a high-fidelity CMS. Clinical data of the infants were: gestational age: 29 (25-41) weeks; birthweight: 1200 (500-3675) g, postnatal age: 6 (2-46) h. METHODS: Statistical analysis was performed with the paired Student's t-test. Multiple regression analysis was used to determine the influence of birthweight and height of the blood pressure on the results. MEASUREMENTS AND MAIN RESULTS: In 51 infants, 255 paired values of SAP, DAP and MAP were recorded. In all recordings, we determined the relationship between SAP, DAP and MAP, using the equation: MAP = alpha%(SAP - DAP) + DAP. For SAP, DAP, MAP and alpha, we computed mean differences (bias) and the limits of agreement (precision). Biases for SAP, DAP, MAP and alpha were significantly different from zero (P < 0.001) and the limits of agreement for SAP, DAP and MAP were wide: 18.8 mmHg, 17.2 mmHg and 15.2 mmHg respectively. The relationship between invasive and noninvasive values is only partly (7-19%) influenced by the height of the blood pressure; low values of SAP, DAP and MAP tend to give overestimated oscillometric values. In the relationship between SAP, DAP and MAP, alpha was found to be 47% invasively (as generally found in the radial artery in newborns) and 34% noninvasively (as generally found in the brachial/radial artery in adults). CONCLUSIONS: Inaccuracy of the oscillometric device may be partly explained by the incorporation of an inappropriately fixed algorithm for final ABP determination in newborns. Care should be taken when interpreting the oscillometrically derived values in critically ill newborn infants. PMID- 8727440 TI - Pain relief with low-dose intravenous clonidine in a child with severe burns. AB - The case of an 11-year-old boy who suffered second and third degree burns to 78% of his body is reported. The large doses of morphine used as analgesia resulted in severe side effects: ventilatory dependence, impairment of gastrointestinal function and psychological disturbance. Intravenous lignocaine was added without benefit. The addition of low-dose intravenous clonidine, however, precipitated a dramatic reduction in morphine consumption with an attendant improvement in ventilatory, gastrointestinal and psychological functions. PMID- 8727441 TI - A new side effect of inhaled nitric oxide in neonates and infants with pulmonary hypertension: functional impairment of the neutrophil respiratory burst. AB - INTRODUCTION: Inhaled nitric oxide (NO) may be beneficial in the treatment of pulmonary hypertension, both of the newborn and in the adult respiratory distress syndrome. Up to now, serious systemic side effects have not been reported. OBJECTIVE: The effect of inhaled NO on superoxide anion production by neutrophils. DESIGN: Prospective study of a consecutive series of 15 neonates and infants. SETTING: Neonatal and paediatric ICUs with a total of 17 beds (university hospital). MEASUREMENTS AND RESULTS: Superoxide anion production was determined by a flow cytometric method using dihydrorhodamine 123 (DHR) as an oxidative probe after the priming of neutrophils with N-formyl-methionyl- leucylphenylalanine (fMLP) or with Escherichia coli. The generated fluorescence was expressed as relative fluorescence intensity (RFI). Inhalation of NO for more than 24 h reduced the superoxide anion production by neutrophils stimulated with E. coli to below baseline values before NO inhalation (mRFI = 158 +/- 25 vs 222 +/- 24; P = 0.03). This decrease was more pronounced after more than 72 h (mRFI = 133 +/- 17). At this time, superoxide anion production by fMLP-stimulated neutrophils was also decreased (mRFI = 40 +/- 3, vs 57 +/- 5; P = 0.03). The reduced capacity of superoxide production persisted throughout therapy with NO and lasted up to more than 4 days after the end of NO inhalation. CONCLUSION: The results suggest that inhalation of NO in patients with pulmonary hypertension causes reduced superoxide anion production by neutrophils stimulated with E. coli or with fMLP. To determine the clinical importance of this systemic side effect with respect to bacterial infections, a randomized controlled study is necessary. PMID- 8727442 TI - The outcome of children admitted to intensive care with meningococcal septicaemia. AB - OBJECTIVE: To review our experience of children with meningococcal septicaemia, and to validate, in our group, severity scores used in different populations to predict outcome. DESIGN: Retrospective review of case notes and charts. PATIENTS: A total of 35 children were admitted to the paediatric intensive care unit (ICU) in the Royal Children's Hospital (RCH) in the 8 years between January 1985 and December 1992 with proven meningococcal septicaemia. RESULTS: Ages ranged from 4 months to 16 years, with a median age of 20 months. The median meningococcal score was 4 and the median PRISM score was 20, with scores above these being significantly associated with death (P < 0.0001). Thirty-two children (91%) received infusions of colloid for hypovolaemia and twenty-nine (83%) received one or more inotropic drugs. Twenty-one children (60%) required mechanical ventilation for a median of 16.5 h (range 7-574). Seven children (20%) underwent plasmapheresis. Six children (17%) underwent haemofiltration and two (6%), peritoneal dialysis. One patient received extracorporeal membrane oxygenation (ECMO) because of circulatory failure. Twenty-one children (60%) developed disseminated intravascular coagulation, renal failure and/or skin or limb necrosis. The overall survival was 66%, and all survivors are functionally normal. CONCLUSION: The mortality from the disease remains at 34% despite the technological advances in intensive care. The PRISM and meningococcal scores are useful in predicting outcome. Novel methods of treatment (e.g., plasmapheresis or ECMO) may be valuable. PMID- 8727444 TI - Intra-abdominal pressure can be measured by measuring the pressure within the stomach. PMID- 8727443 TI - Inspiratory pressure/maximal inspiratory pressure: does it predict successful extubation in critically ill infants and children? AB - OBJECTIVE: To evaluate the accuracy of the initial negative inspiratory pressure (PI) to maximal negative inspiratory pressure (PImax) ratio in predicting extubation outcome for intubated infants and children. DESIGN: A prospective study. SETTING: Pediatric intensive care unit. PATIENTS: A sample of 50 stable intubated pediatric patients who were judged clinically ready for extubation. METHODS: Using a one-way valve, PI and PImax were measured in all patients, after which the < or = ratio PI/PImax was calculated and its accuracy in predicting extubation outcome evaluated. MEASUREMENTS AND RESULTS: A total of 39 patients (78%) were successfully extubated and 11 patients (22%) were not. The mean PI/PImax ratio was not significantly different between extubation successes (0.36 +/- 0.14) and failures (0.45 +/- 0.1) (P > 0.05). The cut-off value of 0.3 for PI/PImax identified in adult patients did not discriminate between extubation success and failure in children. Furthermore, a discriminatory cut-off value other than 0.3 could not be identified for infants and children. CONCLUSION: The PI/PImax ratio cannot be used to predict extubation outcome in pediatric patients. Indices that predict extubation outcome in adults should not be extrapolated to infants and children before testing and validation. PMID- 8727445 TI - Respiratory failure and intensive care treatment in bone marrow-transplanted patients. PMID- 8727446 TI - Rapidly reversible acute cor pulmonale after intravenous injection of crushed dextromoramide (Palfium) pills. PMID- 8727447 TI - Gastrobronchial fistula: report of an unusual case. PMID- 8727448 TI - Microbial contamination of intravenous and arterial catheters. PMID- 8727449 TI - Variations in the structure of neutral sugar chains in the pectic polysaccharides of morphologically different carrot calli and correlations with the size of cell clusters. AB - Carrot (Daucus carota L.) embryogenic callus (EC) loses its embryogenic competence and becomes non-embryogenic callus (NC) during long-term culture. With the loss of embryogenic competence, the cell clusters become smaller and the extent of intercellular attachments is reduced. Pectic fractions prepared from EC and NC were separated into two subfractions by gel filtration. A difference in sugar composition between EC and NC was found only in the high-molecular-mass (ca. 1300 kDa) subfraction, and the ratio of the amount of arabinose to that of galactose (Ara/Gal) was strongly and positively correlated with the size of cell clusters in several different cultures. From the results of sugar-composition and methylation analyses, and the results of treatment with exo-arabinanase, models of the neutral sugar chains of pectins from Ec and NC are proposed. Both neutral sugar chains are composed of three regions. The basal region is composed of linearly linked arabinan 5-Araf) moieties in both types of callus. The middle galactan region is composed of 6-linked galactose, some of which branches at the 3 and 4 positions, and this region is larger and more frequently branched in NC than in EC. Finally, the terminal arabinan region is composed of 5-linked arabinose, branched at the 3 position, and the size of the terminal arabinan is larger in EC than in NC. The significance of the neutral sugar chains of pectins in the interaction of cell wall components and intercellular attachment is discussed. PMID- 8727450 TI - Cervical spine injuries from high-velocity forces: a pathoanatomic and radiologic study. AB - The detailed analysis of the radiologic and pathoanatomic data from 10 human cadaver head-neck complexes defined the type and extent of expected cervical spine injuries after high-velocity flexion-compression loads to the cranium. All specimens demonstrated multiple injuries with both contiguous and noncontiguous patterns. Although all preparations showed evidence of axial compression, a multiplicity of other force vectors, including noncontiguous occurrences of flexion, extension, and shear, were documented. These findings indicate that the injury pattern is not a sequential process but a reaction to changes in the segmental interrelations of the various vertebral column components, including varying vector applications of injurious forces at the segmental level. The presence of moderate or severe spondylotic alterations restricted the distal transmission of injury forces with the principal injury patterns occurring at or proximal to the initial level of severe spondylotic involvement. These data emphasize the need for increased awareness of the presence of multiple cervical spine injuries, both contiguous and noncontiguous, and that separate levels of compromise may not share similar mechanisms of injury. PMID- 8727451 TI - Evaluation of pedicle screw insertion monitored by intraoperative evoked electromyography. AB - The insertion of pedicle screws monitored by evoked electromyography (EMG) was prospectively evaluated in the 132 consecutive patients. The technique involved constant-voltage stimulation and was statistically evaluated at both the arbitrary 20- and 40-V settings. The patients were postoperatively evaluated clinically and radiographically. Computed tomography (CT) scanning was performed for new neurologic deficits. Results were divided into three groups: type 1, a negative EMG response; type 2, a positive EMG response, but no corrective action taken; and type 3, a positive EMG response and corrective action undertaken. Nonparametric statistics were used to evaluate the results at both the 20- and 40 V settings. In the type 3 group, in which corrective action was undertaken, there were no neurologic injuries or screw removals, a statistically significant result. Looking at the two intensity levels, at 20- and 40-V settings, there were no statistically significant differences in the three classifications at either intensity level. We concluded the evoked EMG for monitoring pedicle screw insertion is an efficacious adjunct. A positive response at < 20 V with the constant-voltage technique warrants corrective action. PMID- 8727452 TI - Comparison of female geriatric lumbar-extension strength: asymptotic versus chronic low back pain patients and their response to active rehabilitation. AB - We compared lumbar-extension strength between healthy asymptomatic geriatric females (HEAL) and symptomatic geriatric females (INJ) seeking medical attention for chronic low back pain. The INJ group used the MedX lumbar-extension machine to perform isotonic exercises two times per week and were eventually reduced to one time per week. Range of motion (ROM) and strength were significantly different between groups before beginning the program. After the program, ROM and strength improved significantly and were not different from those of the HEAL group. The average length of treatment was 97 days and 20 visits. Subjective pain ratings were significantly reduced (60%) and exercise weights significantly increased (71%). This reconfirms the notion that many back pain sufferers have weaker lumbar-extension strength and that some symptomatic geriatric women can increase strength with progressive resistance exercise, which leads to a decrease in low back pain. PMID- 8727453 TI - Inverse relation between vertebral body deformity and intervertebral disk narrowing in lumbar spine of elderly women. AB - To investigate the relation between vertebral body deformity and disk narrowing in the lumbar spine of elderly patients, a longitudinal radiographic study of 56 aged women was retrospectively performed. We measured disk areas and body heights on two lateral lumbar radiograph sets obtained with an average interval between examinations of 10 years. The absolute values of disk areas on first examination and the rates of change of area of disks adjacent to nondeformed bodies were smaller than the corresponding values for the deformed bodies. Conversely, deformed bodies tended to have normal or expanded disks regardless of the mode of deformity. Despite the small number of subjects studied, our findings suggested that an inverse relation may exist between vertebral body deformity and disk narrowing. PMID- 8727454 TI - Clinical appearance of contained and noncontained lumbar disc herniation. AB - In a prospective and consecutive study, we evaluated the incidence of common symptoms and neurologic disturbances in 200 patients operated on because of lumbar disc herniation by using a computer-coded protocol with pre- and perioperative registration. The preoperative occurrence of pain at rest, at night, and on coughing was registered. Use of analgesics and walking ability were registered as category data. At examination, a straight-leg-raising (SLR) test was graded in four categories, and results from neurologic findings were collected. At surgery, disc herniation was classified as extruded/sequestered herniation, prolapse, or focal protrusion. There were no significant differences concerning pain at rest or at night related to type of herniation. Pain on coughing was more common in extruded/sequestered herniations. Use of analgesics as well as severe reduction of walking capacity were significantly more common in patients with extrusion/sequestration. The highly restricted SLR test, as well as the crossed positive SLR test, were also significantly more common in patients with extruded/sequestered herniation, and this was also true for the incidence of relevant reflex/extensor hallucis longus (EHL) and sensory disturbance. In conclusion, the clinical appearance of lumbar disc herniation was most "aggressive" in extruded and sequestered disc herniation. The symptoms and signs in disc protrusion were less severe, whereas patients with prolapse had an "intermediate" appearance concerning symptoms and signs. The differences in incidence of common signs in noncontained versus contained herniation were statistically significant; these differences may be of clinical interest for patient selection and information as well as in pathophysiologic considerations. PMID- 8727455 TI - Important numbers in persistent low back pain. PMID- 8727456 TI - Persistent back pain and sciatica in the United States: patient characteristics. AB - Low back pain is an extremely common, seriously disabling, nonfatal public health problem worldwide. The National Low Back Pain Study was a multicenter study of a large, heterogeneous group of patients who have been referred to either a neurosurgeon or an orthopedic surgeon for the evaluation and treatment of a persistent complaint of low back pain. In this paper, we characterize persistent low back pain patients and their complaints, describe the impact of persistent low back pain on the patients' functional and psychological status, report on the patients' medical characteristics, and identify treatments that are currently prescribed for these patients. Persistent low back pain is most common among people in their mid-to-late thirties and early-to-mid forties. The patients are mostly white, well educated, and generally affluent. The majority are gainfully employed, but some quit working because of pain and those who do tend to be less educated, and more likely to be involved in litigation. The average patient has had low back pain intermittently for 10 years. The pain is usually well localized but its severity varies considerably. Besides pain, most persistent low back pain patients report a variety of motor and sensory deficits. Patients also report significant functional impairment at work, at play, and at home. The typical patient does not, however, display significant psychological distress. Most patients have consulted multiple health care providers, have received a variety of treatments, and have used a variety of medications to alleviate pain; a few have been subjected to more aggressive treatment measures including surgery, intradiscal therapy, and narcotic and psychoactive drugs. None of these treatments has been effective. Physical examinations of these patients do not provide significant clues for making a definitive diagnosis. Nonspecific abnormalities such as muscle spasm, tenderness, and trigger points are quite common, but motor weakness and sensory deficits in the lower extremities, and reflex changes in the knees and ankles, are much less common. The classic combination of reflex changes, motor weakness, and sensory deficits associated with specific protruded discs are extremely rare even though one of three patients had a diagnosis of disc herniation. Diagnostic imaging studies revealed that the majority of persistent low back pain patients have spondylotic abnormalities involving root compression or lumbar instability or both, with root compression as the primary cause of the complaint. Myofascial syndrome and lumbar instability were the next most common diagnoses. After a thorough evaluation by specialists in spinal disorders, three of five persistent low back pain patients were prescribed an additional course of conservative therapy, one of five was prescribed surgery, and the rest were prescribed no treatment. Persistent low back pain patients appear to be a distinct group of low back pain patients who are different from patients who have similar nonpersistent acute symptoms and those who have the chronic pain syndrome characterized by significant behavioral and psychological co-morbidities. PMID- 8727457 TI - Lateral intertransverse process single-level fusion for salvage of the unstable failed posterior lumbar interbody fusion. AB - Our study was an attempt to salvage severely unstable posterior lumbar interbody fusions (PLIFs) by using the lateral bilateral intertransverse single-level (floating) fusion. A retrospective review of six patients with failed L4-L5 PLIFs was carried out. These patients underwent reoperation between 2 weeks and 2 years after the initial surgery. All patients had persistent symptoms, including low back pain and sciatica, interfering with activities of daily living. Computed tomography (CT) scans revealed extrusion of the bone plug into the spinal canal in five patients. All six had a nonunion of the PLIF, and three patients had grade 3 or 4 iatrogenic spondylolisthesis caused by the PLIF. The operation involved decompression followed by lateral intertransverse process fusion with autogenous bone graft. All patients were placed in external immobilization after the procedure for 3 months. Follow-up averaged 38.5 months (range, 21-55), at which time five had substantial pain improvement. Five were radiographically fused. One had a second nonunion, which subsequently healed after reoperation with repeat lateral fusion and Luque plate hardware. Our fusion rate was 83% without the use of instrumentation. In conclusion, we recommend the simplistic procedure of lateral bilateral intertransverse process single-level fusion along with liberal external immobilization for the difficult problem of salvage of failed PLIFs. PMID- 8727458 TI - Cervical spine injury in a boxer: should mandatory screening be instituted? AB - Cervical spine fracture is a rare, yet potentially catastrophic complication associated with boxing. Neurologic deficits, ocular injuries, or other traumatic musculoskeletal injuries have been commonly reported. Symptoms of cervical spine injury may be minimal or absent. Cervical spine fracture may be undiagnosed and unreported in boxers with presumed soft-tissue injury to the head and neck. We describe a young athlete who sustained a transient spinal cord injury while boxing, which required a cervical spine fusion and postoperative immobilization in a halo vest. This athlete also had an os odontoideum, which placed him at significant risk for such an injury. Pre- and postparticipation screening of the cervical spine should be considered in all boxers. PMID- 8727459 TI - Diskitis associated with transpedicular screw fixation. PMID- 8727460 TI - Lumboperitoneal shunt for cauda equina syndrome in ankylosing spondylitis. AB - Cauda equina syndrome is a rare complication in the late stage of ankylosing spondylitis, for which approximately 60 cases have been reported in the literature. The cause of the syndrome is unclear, and there is no effective treatment. Recently lumboperitoneal shunt was reported to have been effective in two patients. In our study, we performed lumboperitoneal shunt in a patient and evaluated the condition after the operation compared with that preoperatively. Some alleviation of neurologic symptoms was observed for 6 months after operation. Histopathologic examination of the dural diverticulum revealed a residual change after old inflammation. Lumboperitoneal shunt was an effective surgical treatment for cauda equina syndrome in this patient with ankylosing spondylitis, but its effects were not extreme. Arachnoiditis is suggested to be involved in the pathogenesis of cauda equina syndrome. PMID- 8727461 TI - Post-traumatic progressive cervical myelopathy in patient with congenital spinal stenosis. PMID- 8727462 TI - Postgraduate medical training for deck officers. AB - 347 Finnish deck officers completed the questionnaire on medical training, knowledge and skills. The following conclusions could be drawn: a. Medical training must be based on generally accepted standards, both nationally and internationally. b. More practical exercises should be included in the training. c. Refresher medical training clearly increases knowledge and skills but it also gives the possibility to train, maintain and repeat practical routines. d. Evaluation of the skills should be a part of qualification. e. Good medical knowledge on board ship needs radio-medical services and vice versa. PMID- 8727463 TI - Evaluation of exposure to noise in seafarers on several types of vessels in Polish Merchant Navy. AB - Noise was measured on merchant ships of 3 types, during routine sea voyages. It was found that the equivalent noise levels calculated for engine crew members exceeded the accepted hygienic norm by 1-2 dB. Also audiometric examinations at the beginning and on completion of a voyage were performed, enrolling seamen from the engine room and from the deck crews. They showed statistically significant differences of temporary shift of the hearing threshold, more pronounced in the engine room crew. PMID- 8727464 TI - Knowledge of seafarers about AIDS problems and their vulnerability to HIV infection. AB - The results of a questionnaire survey conducted among 243 seafarers in Rijeka in 1989-1990 indicated that the knowledge of ratings among them on AIDS and HIV virus transmission was inadequate. The majority of them had not used condoms for protection against HIV virus infection. More education on AIDS should be given to seafarers, particularly those employed on ships flying flags of developing countries. PMID- 8727465 TI - Selected problems of health and safety of divers in the light of Polish regulations. AB - The work addresses legal aspects of employing amateurs (diving clubs members) at underwater works, and lack of standards and regulations regarding light diving equipment for diving teams. New legislation is needed to lay down pensionable age for divers, regulate the employment of women for underwater works in some professions, and medical aspects of fitness certification for divers. PMID- 8727466 TI - Evaluation of the risk of decompression sickness in saturated nitrox diving by examination of haemostasis. PMID- 8727467 TI - Long-term monitoring and prevention of occupational lead exposure in a production plant. AB - During 19 years of monitoring and prevention of occupational lead exposure in a production plant the authors studied lead emission sources and exposure intensity encountered in the work place, the indices of current and long-term lead exposure, and markers of toxic effects of lead in 132 persons examined together 935 times. Lead poisoning was confirmed in 20 persons, i.e. about 2 per cent of examinations, whereas in 12 per cent signs of enhanced lead absorption were found. Based on a complex simultaneous assessment of many parameters, appropriate prophylaxis and therapy were applied to lessen the toxic effects of lead exposure. It is pointed out that technical improvements are fundamental for limiting lead emission and absorption in the work place. As it has been demonstrated, the proper cooperation between the physician, the toxicological laboratory, occupational safety service and the plant administration effectively contribute to the limitation or even elimination of lead exposure, thus reducing the possible future health effects of long-term lead exposure. PMID- 8727468 TI - Thermal desorption-gas chromatography for the determination of emission of volatile organic compounds from furnishing materials. AB - In order to determine volatile organic compounds (VOC) released from the building and furnishing materials gas chromatography with air samples enrichment on Tenax TA and thermal desorption was used. The results obtained were compared with the results of air samples enrichment on active carbon with carbon disulfide extraction. To the thermal desorption the home made device, a similar to device developed at Research Triangle Institute (USA), was used. The both methods of the air samples enrichment with thermal desorption or carbon disulfide extraction showed that the emission of the volatile organic compounds from the PVC floor covering is very low within the range from 0.01 to 0.03 mg/m2.h. Thermal desorption technique allows to detect a larger number of compounds in trace levels. PMID- 8727469 TI - Effect of environmental conditions on re-emission of formaldehyde from textile materials. AB - Furnishing textile materials may be a source of formaldehyde (CH2O) re-emission if this compound is present in the environment. In the examinations performed the effect of temperature (23 degrees C, 35 degrees C) and air humidity (RH: 45%, 85%) upon CH2O re-emission from curtain materials was determined. The source of CH2O emission was particle board. CH2O emission was determined in small glass chambers (13.08 dm3) in controlled conditions, one air exchange per hour at loading factor of 1 m2/m3. The results of these examinations showed that the effect of temperature and humidity upon CH2O adsorption and desorption was different. Increase of temperature from 25 degrees C to 35 degrees C caused increase of the magnitude and rate of CH2O re-emission and the phenomenon intensified at high air humidity. PMID- 8727470 TI - Verapamil influences the effect of nonionic contrast agent on platelet membrane structure and aggregation. AB - Anticoagulant and antiplatelet function of NICM is weak, thereby presenting a hazard of coronary thrombus growth in patients subjected to coronary angiography. Similar antiplatelet effect was demonstrated for verapamil (V). The present study was designed to determine iopromide induced modifications in blood platelet membrane structure and aggregation in patients with primary hypertension and ischaemic heart disease receiving V compared to non-receivers of V. The blood for examinations was collected by Judkins' catheter placed in the vicinity of the coronary vessel ostium, and next centrifuged to obtain platelet-rich plasma. The ADP- and ristocetin (R)-induced blood platelet aggregation was determined by means of an aggregometer while their membrane structure was studied by electron paramagnetic resonance. The spin-label used, 4-maleimide-2, 2, 6, 6-tetramethyl piperidine-1-oxyl (MSL) binds covalently to the -SH and -NH2 groups of platelet membrane proteins. The modifications in various spectral components, examined by means of a spectrometer, reflect conformational changes in these proteins. In non receivers of V, iopromide diminished the aggregation, significantly in the case of ADP (p = 0.05), whereas in the receivers of V the iopromide-induced platelet aggregation examined with ADP did not change while with R was even enhanced (p = 0.05, Fig. 2). The platelet membrane protein conformation also changed due to iopromide: in non-receivers of V the accessibility of the above named groups for MSL was significantly reduced, p < 0.05, but in the receivers of V--inversely- significantly increased, p < 0.05. The results obtained confirm that in non receivers of V, iopromide modifies the conformation of platelet membrane proteins which, in turn, may result in the lessening of the aggregation rate, favourable as regards the conditions of coronary thrombus growth. The tendency in the receivers of V was reverse and, however its nature remains obscure, this effect should be an indication for discontinuing the use of V several days before subjection the patient to coronary angiography. PMID- 8727471 TI - Surface distribution of the coliform bacteria in the Gdansk Bay. AB - In 1993, during the research cruises 224 samples of sea water were taken from the stationary samples points. The most probable number of faecal coliform bacteria was determined for each sample. The results were shown on maps. The maps show ranges and directions of pollution expansion, indicate places of contamination input and confirm that the weather conditions and the shape of coastal line have an effect on pollution distribution in Gdansk Bay. The obtained results show that range of the bacteriological contamination carried into the Gdansk Bay from land (as municipal sewage, industrial waste and rivers) is limited to narrow coastal zone. This knowledge is of great importance when estimating the sanitary state of holiday sea resorts and also allows to plan the activities which tend towards reduction of the sewage inflow through the separation of local pollution sources. PMID- 8727472 TI - Peak oxygen consumption and ventilatory thresholds on six modes of exercise. AB - In order to compare responses on six modes of exercise for maximal oxygen consumption (VO2peak) and ventilatory thresholds (VT-1, VT-2), 10 male recreational exercisers (23 +/- 3 yrs) completed incremental maximal tests on treadmill, stationary skier, shuffle skier, stepper, stationary cycle, and rower. After extensive habituation, VO2peak, VT-1, and VT-2 were determined during each maximal bout. A MANOVA followed by ANOVAs, Tukey post hoc tests, and noncentral F tests indicated that the treadmill elicited a significantly higher peak oxygen consumption than did the other modes, and the skier and stepper values were higher than the rower. VO2 at VT-1 was higher on the treadmill than cycle. The treadmill also elicited a higher VO2 at VT-2 than the shuffle skier, cycle, and rower. However, no differences were observed among modes for VT-1 and VT-2 when expressed as a percentage of VO2peak. These results suggest that the treadmill elicits a higher aerobic capacity measure than other modes, but the ventilatory threshold responses (% VO2peak) are similar among modes. PMID- 8727473 TI - Heat strain in protective clothing following hot-wet or hot-dry heat acclimation. AB - The purpose of the present study was to compare the heat strain while wearing nuclear, biological, and chemical (NBC) protective clothing following a hot-wet (HW) or hot-dry (HD) heat acclimation protocol. Twenty-two males were assigned to groups HW (n = 7), HD (n = 8), or control (C, n = 7). Subjects were evaluated during continuous treadmill walking while wearing lightweight combat clothing and during intermittent exercise while wearing the NBC protective clothing. While wearing Combat clothing, greater decreases in rectal temperature (Tre), mean skin temperature (Tsk), and heart rate were observed for both acclimation groups. For the NBC clothing trials, lower Tre, Tsk, and heart rates were observed only for group HW. The time required for Tre to increase 1.0 degrees C and 1.5 degrees C was significantly delayed for groups HW and HD. Sweat evaporation increased for HW, whereas no change was found for HD. The most significant changes in Tre, Tsk, and heart rate while wearing the NBC protective clothing occur following heat acclimation that involves wearing the clothing during exercise. PMID- 8727474 TI - Reliability and validity of a portable metabolic measurement system. AB - The purpose of this study was to assess the reliability and validity of a portable metabolic system (TEEM 100) during submaximal and maximal (VO2max) exercise using a computer-based metabolic system as the reference system (REF). Between repeated trials of submaximal exercise at three constant loads, differences in ventilation (Ve) and oxygen consumption (VO2) were 0.2 +/- 4.9 L . min-1 and 0.03 +/- 0.10 L . min-1 for REF, and 1.9 +/- 0.7 L . min-1 and 0.00 +/- 0.17 L . min-1 for TEEM 100. Pooled intraclass reliability coefficients for Ve and VO2 calculated from the repeated submaximal trials were r = .89 and r = .94 for REF, and r = .86 and r = .94 for the TEEM 100. Respiratory exchange ratio (RER) measured by the TEEM 100 was significantly higher (p = .01) at only the lowest workload. At VO2max, the TEEM 100 recorded significantly higher values for FeO2 (p = .01) and RER (p < .001). These results suggest that the TEEM 100 provides reliable and valid measurements of VO2 during submaximal and maximal exercise. PMID- 8727475 TI - Metabolic response of forearm muscle to graded exercise in type II diabetes mellitus: effect of endurance training. AB - In this study, 31P nuclear magnetic resonance spectroscopy was used to monitor muscle metabolism in Type II diabetic subjects (n = 10) during an incremental exercise test. Also the exercise responses of diabetic subjects (n = 4) following submaximal endurance training were assessed and compared to healthy controls (n = 5). Responses to incremental exercise in the diabetic subjects were consistent over time despite minor fluctuations in metabolic control. In the diabetic and control groups, after 12 weeks of training the forearm flexor muscles, power output at the intracellular threshold of acidosis (IT) increased (p < .01) similarly: T0 versus T12: 0.90 +/- 0.09 versus 1.20 +/- 0.13 and 1.03 +/- 0.07 versus 1.22 +/- 0.10 W, respectively. Minimum intracellular pH reached at peak exercise was unchanged after training. The control group, however, became more acidic versus the diabetic group (p < .05) in response to progressive exercise. This difference was maintained over time. Endurance training elicited similar adaptations in forearm muscles of Type II diabetic and control subjects, although there were differences between the two groups in intracellular pH during exercise. PMID- 8727476 TI - Design and evaluation of a modified underwater cycle ergometer. AB - An underwater cycle ergometer was designed consisting of an aluminum cycle frame in water connected with a 1:1 gear ratio to a mechanically braked standard cycle ergometer supported above the water. Three progressive maximal exercise tests were performed (n = 10): (a) the underwater ergometer in water (UEW), (b) underwater ergometer in air (UEA), and (c) a standard cycle ergometer in air (SEA). At submaximal power outputs, oxygen consumption (VO2) and heart rate (HR) were generally lower in the SEA condition (p < .05), indicating that exercise in the upright position was more efficient. Exercise in water (UEW) resulted in lower total exercise duration, maximal HR, and maximal Tes than in air conditions. The upright position (SEA) resulted in greater total exercise duration and maximal power output than the semirecumbent positions. Because of positional differences between the standard and underwater ergometers, air-water comparisons should be made by using the underwater ergometer in water and on land. PMID- 8727477 TI - Seated versus standing cycling in competitive road cyclists: uphill climbing and maximal oxygen uptake. AB - Seven competitive road cyclists (M +/- SE = 23.7 +/- 1.5 yr, 70.5 +/- 1.7 kg) participated to determine the effects of cycling body position on physiological responses during uphill cycling and maximal oxygen uptake (VO2max). There was no significant difference in VO2max between seated and standing positions on a cycle ergometer (66.4 +/- 1.6 vs. 66.4 +/- 1.7 ml . kg-1 . min-1). When the subjects rode their own bicycle on a treadmill, oxygen uptake and heart rate were significantly (p < 0.05) higher during standing when subjects bicycled at 20.0 km . h-1 (4% grade), but no difference was observed when riding at 12.3 km . h-1 (10% grade). Leg RPE was significantly (p < 0.05) lower for standing position up a 10% grade. The results suggest that the standing position is less economical during moderate hill climbing, but during steep hill climbing, it results in a decreased sensation of effort in the legs. PMID- 8727479 TI - Mapping a conserved conformational epitope from the M protein of group A streptococci. AB - The carboxyl terminus of the M protein of group A streptococci (GAS) is highly conserved and contains epitopes that have been shown to induce opsonic antibodies and protection against GAS infection. This region of the protein can also stimulate T cells, which can react in vitro with heart antigens. Since different segments of the carboxyl terminus may be involved in immunity to GAS and in the pathogenesis of autoimmune disease (rheumatic heart disease), it is important to precisely define critical epitopes. However, the M protein is known to be a coiled coil, and a critical immunodominant antibody-binding epitope within this region (peptide 145, a 20-mer with the sequence LRRDLDASREAKK-QVEKALE) is shown here to be conformational. Thus, small synthetic overlapping peptides of 8-12 amino acids in length that span peptide 145 (p145) were unable to capture antibodies present in p145-immune mouse sera or in endemic human sera, even though antibodies raised to these small peptides coupled to diphtheria toxoid could bind the smaller peptides and, in some cases, p145. A series of mutated peptides in which every residue of p145 was sequentially altered also failed to identify critical residues for antibody binding. We thus devised a strategy to produce chimeric peptides in which small peptides copying the M protein sequence were displayed within a larger 28-mer peptide derived from the sequence of the GCN4 leucine zipper DNA binding protein of yeast. A 12-amino-acid window of the p145 sequence was inserted into the GCN4 peptide in such a way as to preserve any potential helical structure. The window was moved along one residue at a time to give a series of peptides representing p145. Circular dichroism demonstrated that these larger chimeric peptides and p145, but not a shorter 12-mer peptide, displayed alpha-helical potential in 50% trifluoroethanol. Certain chimeric peptides efficiently captured antibodies specific for p145 and thus enabled us to map the minimal antibody-binding sequence. RRDLDASREAKK, referred to as J(1)2. The chimeric peptide containing this sequence, referred to as J2, was able to inhibit opsonization of GAS by human antisera containing anti-peptide 145 antibodies. The T-cell response from p145-immunized responder B10.BR mice to J2 and J(I)2 was much lower than the response to p145 and mapped to a different peptide. PMID- 8727478 TI - Simultaneously synthesized peptides on continuous cellulose membranes as substrates for protein kinases. AB - Sets of peptides with defined sequences, each on a separate spot, were synthesized simultaneously on continuous cellulose membranes (SPOTs membranes), which were originally designed for epitope studies. The applicability of the membrane-bound peptides as substrates for protein kinases was tested using protein kinase A, protein kinase C and casein kinases I and II as model enzymes. We found that the peptide-membrane complexes can serve as kinase substrates. Our results suggest that membrane-bound peptides offer a new potential for the investigation of substrate specificity of protein kinases. An advantage to this method is that there is no need for substrate identification and separation, which is required with high-volume random peptide libraries. Membrane-bound peptides may even form a basis for kinase assays with peptides lacking multiple basic amino acids, required for separation of the substrates in conventional assays. Problems connected with protein kinase substrate specificity can be investigated in any laboratory using the rapid and inexpensive SPOTs technique, as neither costly apparatus nor special experience in peptide synthesis is necessary. PMID- 8727480 TI - Interaction with DNA of oligopeptides related to the Arc repressor. AB - Three different peptides, A13, A14x2 and A20, related to the Arc repressor from Salmonella phage P22, were synthesized. They all contained the 13 N-terminal residues of Arc known to form a beta-sheet interacting with the operator OArc. In the case of A20, the tripeptide Lys-Trp-Lys was added in the C-terminal position because of its propensity to increase affinity to DNA. The interaction of the three peptides with OArc and with other related (OMnt from the same phage) and unrelated oligonucleotides was followed using circular dichroism, filter binding assays and DNaseI protection experiments. While Kd = 10(-9) M for the protein, 8.7 x 10(-5) M and 7.7 x 10(-6) M Kd values were obtained for A14x2 and A20 interacting with OArc. PMID- 8727481 TI - Synthesis, secondary structure and folding of the bend region of lung surfactant protein B. AB - Previous theoretical analysis of the primary structure of lung surfactant protein SP-B indicates a disulfide-linked, hydrophobic midsequence that forms a hairpin like motif. Here, we experimentally investigate the secondary structure of the disulfide-stabilized bend region by synthesizing two 12-residue analogs of the SP B midsequence. The native peptide has the same sequence for residues 35 to 46 as native human SP-B, while, in the second mimic peptide, Leu40 and Val41 were replaced with D-Ser and L-His. Both peptides contain cysteine residues at the N- and C-terminus (Cys35 and Cys46, respectively). Oxidation/reduction experiments with fast atom bombardment mass spectrometry showed mass shifts of approximately 2 daltons, consistent with the oxidized peptides existing in solution as monomers, each with one internal disulfide bond (Cys35-Cys46). Since circular dichroism and Fourier-transform infrared measurements show that both peptides assume turn conformations in structure-promoting solvents such as trifluoroethanol (TFE), a structural model is proposed in which Cys35 and Cys46 are brought in close apposition through an internal bend in the peptide. Consistent with this model are electron spin resonance (ESR) results of the mimic peptide nitroxide spin-labeled at Cys35 and Cys46. For the double spin-labeled mimic peptide in TFE. ESR spectra indicated broadening characteristic of either radical interactions or decreased mobility, or both. Increases in radical interactions for the double spin-labeled mimic peptide would be expected for Cys35 and Cys46 approaching within 14 A in structure-promoting solvents, while decreases in spin-label mobility could be due to the formation of a loop. Based on these observations with peptide analogs, residues 35 to 46 probably form a similar bend in the full-length protein. PMID- 8727482 TI - Chemiluminescent nitrogen detection as a new technique for purity assessment of synthetic peptides separated by reversed-phase HPLC. AB - Reversed phase high-performance liquid chromatography with chemiluminescent nitrogen detection (HPLC-CLND) was used for the quantitative analysis of peptides manufactured by solid-phase peptide synthesis. CLND provided quantitative information regarding the nitrogen distribution of peptide samples following HPLC separation. This technique permits the universal quantitation of the "peptide content" of synthetic peptides in an on-line mode without pre- or post-column derivatization and free of interference from non-nitrogen-containing UV chromophores. This paper will show the utility of this novel technique in measuring the peptide content of crude synthetic proinsulin chain C peptide. A mixture of five reference peptides was analyzed to show the homogeneity of the CLND response and used to determine peptide content. Application for the purified product is also discussed. The detection profiles were acquired in parallel with a UV detector. PMID- 8727483 TI - MARS--multiple automated robotic synthesizer for continuous flow of peptides. AB - We have designed and constructed a multiple automated robotic synthesizer, the MARS. Its novel timing procedure for handling multiple synthetic tasks eliminates unnecessary respite time by keeping the robotic arm in continuous operation. Polypropylene syringes equipped at the bottom with polypropylene frits serve as physically independent reaction vessels. All operations are performed by the robotic arm, which is equipped with a specially designed gripper to hold a syringe and to aspirate and dispense liquid. Typically, the MARS synthesizes concurrently 5 to 15 peptides of different length, and once one peptide is finished it automatically starts the synthesis of the next peptide in the queue, assuring a continuous flow of peptides. PMID- 8727484 TI - Mysteries in the making of the cerebral cortex. PMID- 8727485 TI - Cell lineage and patterns of migration in the developing cortex. AB - Knowledge of cell lineage in the cortex is important for understanding normal development as well as brain malformations. We studied cell lineage in rats by injecting a library of up to 3400 retroviruses, distinguishable by PCR analysis and encoding alkaline phosphatase, at E14-19. Histological analysis at P15 revealed normal cell morphology and allowed identification of about 80% of all labelled cells. PCR amplification of DNA tags allowed clonal analysis. Cortical cells labelled at E15 formed clustered or widespread clones with equal frequency. Clustered clones contained one to four cells within about 1 mm that had similar morphology and laminar location. However, 48% of cortical clones contained multiple cell types with widely different locations (2.1-6.7 mm; mean, 3.8 mm). Widespread clones contained two to four 'subunits' (one to five neurons each), spaced at apparent intervals of 2-3 mm, with each subunit morphologically indistinguishable from a clustered clone. Distinct subunits in the same clone usually differed in laminar location suggesting sequential formation. Clones labelled at E17 contained fewer neurons and up to two subunits. Clustered clones seem to be produced by stationary progenitors, whereas progenitors of clusters may themselves be produced by migratory, multipotential cells. PMID- 8727487 TI - The generation of cellular diversity in the cerebral cortex. AB - We have used retroviral vectors to study cell lineage in the embryonic rat cerebral cortex both in vivo and in dissociated cell culture. We provide evidence that during the late phase of corticogenesis, most precursor cells of the ventricular zone are specified for the production of a single cell type, either neurons or one of the glial cell types. Although specified, the precursor cells that generate neurons can apparently generate both pyramidal and non-pyramidal cells. Earlier stages of development are dominated by a different type of precursor cell with a number of properties that lead us to believe that it is the founding, multipotential precursor cell of the cerebral cortex. We discuss a possible model of cell lineage which unifies these various observations. PMID- 8727486 TI - The cell lineage of neuronal subtypes in the mammalian cerebral cortex. AB - We have studied the lineage relationships of pyramidal and nonpyramidal neurons, the principal neuronal types in the cerebral cortex, using a recombinant retrovirus that carries the gene encoding Escherichia coli beta-galactosidase as a lineage marker. The phenotype of every cell of clones of beta-galactosidase labelled neurons generated by intraventricular injection of recombinant retrovirus in rat embryos at different stages of cortical neurogenesis was identified using light and electron microscopy as well as immunohistochemistry for known markers of neuronal subtypes. We found that clonally related neurons in adult rats showed the same morphological and neurotransmitter phenotypes, suggesting that lineages of pyramidal and nonpyramidal neurons are specified as early as E14, the time of onset of neurogenesis. However, when we followed the development of cortical cell lineages, we noted that a significant number of neuronal clones showed a mixed pyramidal/nonpyramidal cell composition during the first three weeks of life. We suggest that the change in the composition of neuronal clones between the third week of postnatal life and adulthood may either be due to changes in the phenotype of some developing neurons or, more likely, to selective cell death. PMID- 8727488 TI - Factors regulating the differentiation of neural precursors in the forebrain. AB - Precursors from the neuroepithelium of the developing cortex and the adult subventricular zone can be cloned in vitro after stimulation with fibroblast growth factor 2 (FGF-2), and they have the potential to give rise to both neurons and glia. The generation of neurons from these clones can be stimulated by either a factor derived from an astrocyte precursor line, Ast-1, or FGF-1. We have shown that neuronal differentiation stimulated by FGF-1 can be inhibited by diacylglycerol lipase inhibitor and mimicked by arachidonic acid, suggesting that the neuronal differentiation is signalled through the phospholipase C gamma pathway. The sequential expression of FGF-2, followed by FGF within the developing forebrain neuroepithelium, fits with the different functions that the two FGFs play in precursor regulation. We have shown that the precursor response to FGF-1 is regulated by a heparan sulphate proteoglycan expressed within the developing neuroepithelium. Precursors restricted to the astrocyte cell lineage can be stimulated by epidermal growth factor or FGF-2F however, the differentiation into glial fibrillary acidic protein-positive astrocytes appears to require a cytokine acting through the leukaemia inhibitory factor-beta receptor. PMID- 8727489 TI - Emx and Otx gene expression in the developing mouse brain. AB - The homeobox genes Emx1, Emx2, Otx1 and Otx2 are all expressed in the rostral brain of embryos at E10. Their expression domains are continuous regions of the developing brain contained within each other, such that the expression domain of Otx2 contains that of the other three genes, the expression domain of Otx1 contains that of Emx1 and Emx2, and the expression domain of Emx2 contains that of Emx1. The Emx1 expression domain includes the dorsal telencephalon and it has a posterior boundary slightly anterior to that between the presumptive diencephalon and telencephalon, whereas the Otx2 expression domain covers almost the entire forebrain and midbrain. Starting from E10.75, Otx2 expression disappears progressively from the presumptive cerebral cortex, whereas Emx1, Emx2 and Otx1 are expressed in this structure until late gestation. In particular, Emx2 appears to be expressed exclusively in the germinal ventricular zone of the developing cerebral cortex. PMID- 8727490 TI - Guidance of thalamocortical innervation. AB - We propose that a sequence of individually simple mechanisms influences the pattern of thalamocortical innervation, which itself contributes to the determination of regional differentiation of the neocortex. In co-culture, the cortex appears to exert a remote growth-promoting influence on thalamic axons from E15, becomes growth-permissive to axon invasion at about E20 and expresses a stop signal, causing termination in layer IV, from P2-3. This cascade of cortical signals may determine the timing of events in vivo. However, any part of the thalamus will innervate any region of the developing cortex in culture, without obvious preference, suggesting that the topographic distribution of thalamic fibres in vivo does not depend on regional chemospecificity. The initial extension of axons from the cortical preplate and the thalamus starts at about E14, and the topography of both may be influenced by their temporal sequences of outgrowth (chronotopy). The axon arrays meet in the basal telencephalon, after which the preplate scaffold may guide thalamic axons and ensure both their 'capture' within the subplate layer and the establishment of the waiting period. The unusual pattern of innervation in the Reeler mutant mouse supports the hypothesis that thalamic axons grow over preplate fibres to find the waiting compartment. PMID- 8727491 TI - Subplate neurons and the patterning of thalamocortial connections. AB - The patterning of the cerebral cortex into functionally distinct domains relies on the formation of appropriate connections between the thalamus and the cortex during development. To identify the mechanisms that underlie cortical target selection by thalamic axons, we have examined the role of cellular interactions in the formation of connections between the lateral geniculate nucleus (LGN) and the visual cortex during development of the cat visual system. The morphology of LGN axons as they grow towards the visual cortex suggests that interactions within the subplate zone may be important in the development of geniculocortical connections. The requirement for subplate neurons in this process was examined by ablating subplate neurons underlying the visual cortex at various developmental stages. When subplate cells are deleted between E38 and E42, prior to target innervation by LGN axons, these axons fail to select the visual cortex as their correct target and instead grow past it, staying restricted to the white matter. Deletion of subplate cells at later stages, between P2 and P7, does not affect target selection, but instead it prevents the segregation of LGN axons into ocular dominance columns within layer IV of the cortex. The effects of subplate neuron ablation suggest that interactions between thalamic axons and subplate cells are of critical importance in the specification of thalamocortical connections during development. PMID- 8727492 TI - The specificity of interactions between the cortex and the thalamus. AB - The functioning of the adult mammalian cerebral cortex depends critically upon precise interconnections between specific thalamic nuclei and distinct cortical regions. Therefore, one central issue in understanding cortical development is determining the cellular and molecular strategies underlying the specification of thalamocortical projections. We address the role of axon-axon interactions and membrane-bound guidance molecules in the establishment of the development of layer-specific patterns of afferent and efferent cortical connections does not depend upon neuronal activity. We present evidence that activity conveyed by thalamic afferents is required for the elaboration of the columnar specificity of cortical circuits. PMID- 8727493 TI - Molecular contributions to cerebral cortical specification. AB - Evidence is accumulating that decisions of cell fate and commitment to specific regional phenotypes in the cerebral cortex occur through cell interactions that likely begin early in development, perhaps in the proliferative zone. We have focused on the development of the limbic cortex in rats, which includes areas involved in both cognitive and autonomic functions and is marked by expression of the limbic system-associated membrane protein. Transplantation studies show that precursor cells are sensitive to environmental cues which can control expression of area-specific phenotypes, including limbic system-associated protein synthesis and connectivity patterns, but early postmitotic neurons faithfully express traits based on their origin in the donor. We have studied this sensitive period of decision making in vitro. Molecules from the epidermal growth factor family influence dramatically the fate of precursor cells, but only in the presence of matrix molecules. In vivo, both the epidermal growth factor receptor and collagen type IV are expressed in the progenitor cell pool indicating that they can directly affect the initial decisions in differentiation. We suggest that early patterns of gene expression, influenced by environmental cues, are likely to provide a specific framework for subsequent decisions that lead to establishing cortical areas. PMID- 8727494 TI - Plasticity in the development of neocortical areas. AB - Heterotopic transplantation analysis suggests that individual areas of the developing neocortex have the capacity to differentiate many of the architectural and connectional features normally characteristic of other neocortical areas. Many studies indicate a pivotal role for thalamocortical afferents in the differentiation of the area-specific features that distinguish neocortical areas. Both activity-dependent and activity-independent mechanisms contribute to the patterning of thalamocortical afferent terminations. The available evidence suggests that positional information is established in the cortical subplate and that this information controls the precise targeting of developing thalamocortical axons. In this way appropriate thalamocortical relationships can be established that allow these afferents to promote the differentiation of the functionally specialized and anatomically distinct areas of the adult neocortex. PMID- 8727495 TI - The roles of growth factors and neural activity in the development of the neocortex. AB - Previous research on primarily the peripheral nervous system has shown that soluble growth factors help control key developmental events by contributing to dynamic autocrine and paracrine signalling systems. Much less is known about the roles of these substances in neocortical development. Using cell and tissue culture paradigms, we have demonstrated that soluble growth factors are produced by the neocortex and its subcortical targets, and that these tissues can respond to them. There are several possible functions for these factors in neocortical development in vivo: they may initiate axonal growth from neocortical neurons and/or their afferents; accelerate or guide that growth; and/or play a role in the later refinement of connections. Although none of these possibilities can be excluded, the existing evidence strengthens the hypothesis that soluble growth factors are important for the early postnatal growth and refinement of neocortical connections, when their levels of release may be regulated by neocortical activity. At present we do not know which growth factors are involved in these processes, but the results of preliminary experiments indicate that neurotrophins and fibroblast growth factor are prime candidates. PMID- 8727496 TI - Factors that are critical for plasticity in the visual cortex. AB - Factors that may be critical for plasticity in the visual cortex are evaluated according to three criteria. (1) Do antagonists to the factor abolish plasticity? (2) Does the concentration or activity of the factor peak with the critical period for plasticity? (3) Does rearing in the dark, which postpones the critical period, affect the factor in a similar fashion? N-methyl-D-aspartate receptors fulfil all three criteria. Metabotropic glutamate receptors fulfil two of them. Most other putative factors do not fulfil more than one. PMID- 8727497 TI - Cortical development and neuropathology in schizophrenia. AB - Epidemiological studies suggest that perturbations occurring during pregnancy can increase the incidence of schizophrenia among offspring. Examination of the neuropathology of the brains of some schizophrenics suggests that a defect in the later phases of cerebral cortical development, notably the last phases of neuronal migration and the establishment and refinement of patterns of cortical connections, may be involved. Most of these studies are conjectural, and the relationship between primary lesions and potential secondary retrograde and anterograde effects in the circuitry linking the prefrontal cortex, basal forebrain, mediodorsal thalamus and medial temporal cortex is unknown. Our hypothesis, based on neuromorphological and gene expression studies, is that a disturbance of migration or in the pattern of preprogrammed cell death in the subplate zone of the developing cerebral cortex causes a failure to establish normal patterns of connections in the overlying cortex. This compromised circuitry subsequently decompensates, leading to schizophrenic symptoms and activity-dependent manifestations of altered gene expression for neurotransmitter and receptor-related molecules. PMID- 8727498 TI - Pathology of cortical development and neuropsychiatric disorders. AB - Epilepsy is a well-documented consequence of about 150 rare genetic syndromes and malformations of the central nervous system. These syndromes are generally associated with fairly gross defects within the central nervous system and they were thought to be responsible for a small minority of cases. However, improved methods of neuropathological investigations and extensive magnetic resonance imaging studies have revealed a range of disturbances in cortical cytoarchitecture in patients with epileptic seizures previously considered as idiopathic (up to 70% of epilepsy). Structural abnormalities have also been demonstrated in the brain in schizophrenia. These consist of disturbed cortical cytoarchitecture (best described in the temporal lobe) and a diffuse loss of grey matter. The absence of the pathological stigma characteristic of degenerative processes indicates that these structural changes are the result of an abnormal pattern of brain development. The relationship between the type and location of developmental abnormality and the subsequent clinical syndrome (e.g. generalized or localized epilepsy) and the effects of aberrant cortical development on the functional integrity of the adult brain require definition. PMID- 8727499 TI - Visual evoked potential enhancement by an artificial neural network filter. AB - The application of an artificial neural network filter (ANNF) to estimate the visual evoked potential (VEP) is presented. VEP is the gross electrical response of the brain to visual stimuli. Due to the low SNR, it is difficult to extract response from individual stimulus trials. The ANNF we used estimates the deterministic component of the signal and removes the noise uncorrelated with the stimulus, even when the noise is colored. The ANNF is trained through back-error propagation with a data set consisting of a training signal and a target signal. The training signal is the raw VEP from a single trial having a SNR of about -5 dB, while the target signal has a higher SNR which is achieved by ensemble averaging 100 stimulus trials. Simulated signals were generated to test the performance of the ANNF. Results show that the ANNF could greatly enhance the SNR of the VEP to single visual stimulus. Thus the total number of ensembles is reduced. In clinical applications, the traditional ensemble averaging method requires a hundred ensembles to determine the VEP. When ANNF is used, about 20 ensembles are sufficient for the same purpose. PMID- 8727500 TI - Dentin bonding system. Part I: Literature review. AB - Currently, clinicians face choices of restoration including amalgams (mercury based, gallium-based alloys, or mercury-free silver-based substitutes), composite resins, ceramics, and gold alloy. In order to choose an appropriate restorative material, many parameters are involved; they include preparation time requirements, finishing and polishing, marginal integrity, anatomy and contours, chipping and fracture, sensitivity, microleakage, wear resistance, and corrosion resistance. It is generally believed that amalgams are still evaluated as the best of all restorative materials as far as the aforementioned parameters are concerned. It is claimed that the amalgams exhibit in the range of 10 to 25 service years, while the composite resin exhibits ranging between 7 and 11 service years. When a composite resin requires a mass large enough for indirect fabrication, a bonding system is demanded with which this restoration should form an instantaneous, impervious, and stable bond to the tooth structure. Roughly a quarter century has passed since the research and development of a promising dentin bonding system was initiated. We are now in the fifth generation of the dentin bonding system, during the research and development of which various types of bonding agents as well as bonding models have been introduced. In this article, the history of development of the bonding agents and the understanding of the bonding mechanism will be reviewed. PMID- 8727501 TI - Parametric stress analysis of bonded "combination-materials" type of orthodontic brackets. AB - The finite element analysis method and a two-dimensional idealization were used to conduct a detailed parametric study of the stresses in and displacement of models of bonded edgewise "combination-materials" type of orthodontic brackets when subjected to loading and constraint conditions that are deemed to be the same as those that exist in vivo. A "combination-materials" type bracket is herein defined as one in which different materials are used in fabricating the main body and the archwire slot. The present study was conducted in three parts. In Part 1, 16 model cases were analyzed, involving 4 different combinations of materials (for the main body of the bracket and its archwire slot) and 4 different overall bracket configurations (comprising main body, base and slot). The focus of Part 2 was the model bracket configuration that was, on the basis of Part 1 results, deemed to show the "optimum performance". In this part, the parameter investigated was the value of the modulus of elasticity of the adhesive, Eadh. In Part 3, the concept of an efficiency index of the bonded bracket-archwire system, eta, was introduced and explained. The dependence of eta on stated characteristics of a rectangular archwire, for the "optimum performance" model, was explored. Part 1 results led to the conclusion that the "optimum performance" model has equally angulated buccal and lingual edges and the main body and archwire slot are fabricated from glass fiber-reinforced polycarbonate and stainless steel, respectively. Part 2 results showed that, for the aforementioned "optimum performance" model, the longitudinal displacement of the archwire slot is fairly insensitive to Eadh. Part 3 results showed that eta is intimately related to each of the wire characteristics studied (namely cross sectional dimensions, "interbracket distance" and modulus of rigidity of the material). PMID- 8727502 TI - Prevention of platelet adhesion on the polysulfone porous catheter by saline solution perfusion, I. In vitro investigation. AB - A new technique for the prevention of platelet adhesion on the polysulfone (PSf) porous catheter system by means of saline perfusion was investigated. Commercially available ultrafiltration PSf hollow fibers were used as porous catheter tubes. Sixty milliliter of citrated canine blood was circulated for 20 min through the annular catheter flow cell system in vitro. Platelet adhesion onto the PSf porous catheter tube decreased sharply with increased saline solution perfusion rate. From the semilogarithmic relationship between the number of adhered platelets and perfusion rate, the critical perfusion rate for preventing adhesion of platelets could be about 0.3 ml min-1 cm-2. After the surface of PSf porous tubes was modified by non-ionic surfactant coating and air plasma treatment for 20 seconds without saline perfusion, the number of adhered platelets was lower than that of the control tube. These modification methods may be more effective in the prevention of initial platelet adhesion if combined with saline perfusion. PMID- 8727503 TI - Puncture-mediated gene transfer to the skin. AB - Simple and efficient gene transfer to the skin would facilitate many local and systemic gene therapy applications. Here we report a novel approach that allows installation and expression of plasmid DNA without a particulate carrier into the skin for the purposes of gene therapy. A device with a constantly high-frequency oscillating bundle of fine metal needles allows puncturing of the skin and leads to DNA transfer to skin-associated cells and expression of reporter genes in mice. Furthermore, puncture-mediated instillation of an expression vector coding for a single T cell epitope induces specific cellular immune responses. If this approach could be optimized, puncture-mediated gene transfer might be useful for the treatment of large body areas with plasmid DNA for the purpose of somatic gene therapy. PMID- 8727504 TI - Systemic and portal vein delivery of human kallikrein gene reduces blood pressure in hypertensive rats. AB - There is an inverse correlation between systemic blood pressure and urinary kallikrein levels in humans and hypertensive animal models, suggesting that the tissue kallikrein-kinin system plays an important role in blood pressure regulation. In this study, we explored the potential of human kallikrein gene delivery on blood pressure reduction in spontaneously hypertensive rats (SHR). The human tissue kallikrein gene or cDNA was placed under the control of following promoters: the metallothionein gene metal response-element (MRE-pHK), albumin gene (ALB-pHK), Rous sarcoma virus 3' long terminal repeat (LTR) (RSV cHK), and cytomegalovirus (CMV-cHK). A single injection of these kallikrein DNAs results in a significant reduction of blood pressure in SHR, which lasts for 5-6 weeks. Systemic delivery of CMV-cHK, RSV-cHK, and MRE-pHK has a greater effect on blood pressure reduction than ALB-pHK, whereas intraportal vein gene delivery of ALB-pHK is more effective than the other kallikrein DNA constructs. The degree of blood pressure reduction depends on the amount of administered DNA and the age of the animals. Reduction of blood pressure was observed in adult, but not young, SHR. The expression of human tissue kallikrein in rats was identified by an ELISA that is specific for human tissue kallikrein. No antibodies to either human tissue kallikrein or its DNA were detected in rat sera after somatic gene delivery. These results show that somatic gene delivery of human tissue kallikrein causes a lowering effect of systolic blood pressure in genetically hypertensive rats and provide valuable information for kallikrein gene therapy in the treatment of hypertension. PMID- 8727505 TI - Comparison of efficiency of infection of human gene therapy target cells via four different retroviral receptors. AB - The relative efficiency of transduction of gene therapy target cells was measured for retroviruses bearing the envelopes of amphotropic murine leukemia virus (MLV A), xenotropic murine leukemia virus (MLV-X), gibbon ape leukemia virus (GALV), feline leukemia virus subgroup B (FeLV-B), and the feline endogenous virus RD114. These viruses use various cell-surface receptors. Activated peripheral blood lymphocytes (PBL) and primary melanoma cultures were infected relatively poorly by MLV-X pseudotypes. RD114 pseudotypes infected PBL relatively well, whereas bone marrow progenitor cells were efficiently infected by all viruses. Helper free virus bearing the envelopes of MLV-A, RD114, or GALV was similarly tested. All infected melanoma or bone marrow progenitor cells efficiently, whereas MLV-A was relatively inefficient for infection of PBL. The general utility of RD114 pseudotyped virus for gene delivery coupled with its resistance to inactivation by human serum makes this envelope the most suitable choice for in vivo gene therapy. PMID- 8727506 TI - Efficient adenovirus-mediated gene transfer to basal but not columnar cells of cartilaginous airway epithelia. AB - Adenoviral vectors (AdV) developed for treatment of the pulmonary manifestations of cystic fibrosis (CF) can deliver, with high efficiency, transgenes to respiratory epithelial cells grown in culture. This study investigated the efficiency of AdV-mediated gene transfer to murine and human respiratory epithelium in vivo and concluded that the epithelial cells facing the lumen of the respiratory cartilaginous airways (columnar cells) are poorly transduced with AdV. Mechanical injury to the epithelium, however, leads to efficient in vivo gene transfer by exposing a susceptible epithelial subtype (basal cells). Increased gene transfer efficiency in vivo after injury is not a nonspecific response because the proliferative status of the epithelium after injury was shown not to correlate temporally to the increased transduction susceptibility of the epithelium. Although basal cells were the cell type transduced at the time of vector delivery, with time, basal cell differentiation to columnar cells occurred with maintenance of transgene expression. Collectively, these results show that murine and human cartilaginous airways are poorly transduced by AdV. To correct the cartilaginous airway CF bioelectrical defect in vivo, efforts should be directed to increase the tropism of AdV to the columnar airway epithelial cells. PMID- 8727507 TI - Intraluminal water increases expression of plasmid DNA in rat lung. AB - Effective gene delivery to specific organs is a major goal for human gene therapy. The lung's structure allows instillation of agents into the airspaces, directly adjacent to the lung epithelium. We hypothesized that the airspace instillation of hypotonic solutions would increase the permeability of the lung epithelium and increase DNA uptake. This hypothesis was tested by instilling plasmid DNA (p4241) encoding the luciferase gene in isotonic and hypotonic solutions. The highest luciferase expression in the lung was achieved after the instillation of this plasmid DNA in distilled water. Aerosolization of water just before the instillation of the plasmid DNA also enhanced the expression level of luciferase in the lung. In addition, an intralobar instillation of the plasmid DNA in water significantly increased the luciferase expression, suggesting that the instillation of the plasmid over a smaller surface area increased expression. Levels of expression could be measured for 3 days. Water increases the permeability of lung epithelial cells transiently and/or enhances gene expression and can be used to achieve gene expression in the lung airspaces for short intervals without toxicity. PMID- 8727508 TI - Hepatic expression of the catalytic subunit of the apolipoprotein B mRNA editing enzyme (apobec-1) ameliorates hypercholesterolemia in LDL receptor-deficient rabbits. AB - Apolipoprotein (apo) B48, a protein contained in intestinally derived lipoprotein particles, is synthesized by post-transcriptional editing of apoB100 mRNA. This reaction is mediated by an enzyme complex that includes the catalytic subunit, apobec-1. The liver of most mammals, by contrast, contains only unedited apoB mRNA and secretes apoB100, the major protein component of plasma low-density lipoprotein (LDL). Because rabbits, like humans, fail to edit hepatic apoB100 mRNA, we introduced a recombinant adenovirus encoding apobec-1 into the livers of LDL receptor-defective rabbits to determine the impact on lipoprotein metabolism of hepatic apoB48 secretion. Transgene expression was mainly confined to the liver and was sustained for up to 3 weeks following virus administration, as evidenced by the presence of apobec-1 mRNA and the ability of hepatic S100 extracts to edit a synthetic apoB RNA template in vitro. The transient induction of hepatic apoB mRNA editing accompanied alterations in very-low-density lipoprotein (VLDL) size, the presence of apoB48 in fractions spanning the VLDL and LDL range, and modest reductions in total plasma cholesterol levels. PMID- 8727510 TI - Arterial gene therapy for restenosis. PMID- 8727509 TI - Arterial gene transfer for therapeutic angiogenesis in patients with peripheral artery disease. AB - The age-adjusted prevalence of peripheral arterial disease (PAD) in the U.S. population has been estimated to approach 12%. The clinical consequences of occlusive peripheral arterial disease (PAD) include pain on walking (claudication), pain at rest, and loss of tissue integrity in the distal limbs; the latter may ultimately lead to amputation of a portion of the lower extremity. Surgical bypass techniques and percutaneous catheter-based interventions may be used to successfully revascularize the limbs of certain patients with PAD. In many patients, however, the anatomic extent and distribution of arterial occlusion is too severe to permit relief of pain and/or healing of ischemic ulcers. No effective medical therapy is available for the treatment of such patients. The purpose of this clinical protocol is to document the safety of therapeutic angiogenesis achieved in this case by percutaneous catheter-based delivery of the gene encoding vascular endothelial growth factor (VEGF) in patients with PAD; and, as secondary objectives, investigate the bioactivity of this strategy to relieve rest pain and heal ischemic ulcers of the lower extremities. The rationale for this human protocol is based upon preclinical studies performed in a rabbit model of hindlimb ischemia. These studies are described in detail below and in the manuscripts enclosed in the Appendix to this proposal. In brief, a single intra-arterial bolus of VEGF recombinant human protein, delivered percutaneously to the ischemic limb via an intravascular catheter, resulted in angiographic, hemodynamic, physiologic, and histologic evidence of augmented collateral artery development. Subsequently, similar results were achieved using an angioplasty catheter with a hydrogel-coated balloon to deliver 400 micrograms of a plasmid containing the cDNA for VEGF to the internal iliac artery in the same animal model. Accordingly, we propose to administer arterial gene (VEGF) therapy to patients with rest pain and/or ischemic leg ulcers considered not to be candidates for conventional revascularization techniques. The dose of plasmid to be administered will be progressively escalated beginning with 500 micrograms for the first four patients, 1000 micrograms for the following six patients, 2000 micrograms for the third group of six patients, and 400 micrograms for the fourth group of six patients. PMID- 8727511 TI - Modification of tumor suppressor gene expression and induction of apoptosis in non-small cell lung cancer (NSCLC) with an adenovirus vector expressing wildtype p53 and cisplatin. PMID- 8727512 TI - Effect of aluminum on hematopoiesis after intraperitoneal exposure in rats. AB - The experiments were performed on female Wistar rats that received aluminum chloride every day intraperitoneally, 4 mg Al/kg for 3 weeks. In the blood selected morphological factors and delta-aminolevulinic acid dehydratase activity (ALA-D) were investigated. In the kidney and liver ALA-D, delta-aminolevulinic acid synthase (ALA-S), and heme oxygenase activity were also determined. After aluminum chloride administration the most sensitive indicator was an increase of heme oxygenase activity in the liver and a decrease in iron levels in the serum of rats. Aluminum also increased ALA-S activity in the kidney and liver of rats. No changes of ALA-D activity in the liver, the kidney, and the blood were observed. The decreasing of hemoglobin, hematocrit, mean corpuscular hemoglobin mass, and mean corpuscular hemoglobin concentration was noted after 3 weeks of exposure. PMID- 8727513 TI - Effect of dimethoate on body growth of representatives of the soil living mesofauna. AB - For the elucidation of effects of a pesticide on life-cycle parameters of arthropods, applicable in the development of ecotoxicological test procedures with soil living animals, attention was drawn to body growth, which has previously been described as a sensitive parameter in Collembola. Body growth of the following three soil-dwelling arthropods was studied under the influence of dimethoate: Folsomia fimetaria (L.), Hypogastrura assimilis Krausbauer, and Hypoaspis aculeifer Canestrini. The effect of this insecticide and acaricide proved to be sex and species specific. Two models were compared for modeling individual body length. The logistic growth model fitted data for all species, whereas the growth model of von Bertalanffy was less applicable for F. fimetaria and H. aculeifer. The growth rate coefficient of the logistic model proved to be a robust parameter useful for predicting effects of dimethoate on juvenile growth of F. fimetaria and H. aculeifer. Adverse effect concentrations on growth for these species were the following: EC10 = 0.11 mg dimethoate/kg dry soil for F. fimetaria and EC10 = 0.59 mg dimethoate/kg dry soil for H. aculeifer. For H. assimilis, no adverse effects on growth were observed at the tested concentrations. The results are compared to other life-cycle parameters (end points). PMID- 8727514 TI - Measurement and validation of kinetic parameter values for prediction of biodegradation rates in sewage treatment. AB - Analysis of about 140 oxygen uptake curves from standard biodegradation tests with the electrolytic respirometer yielded growth kinetic parameter values. The maximum observed growth rate, mumax, on the 52 different substances varied between 1 and 6/day. The measurement of the half-saturation constant Ks at the depletion phase of the oxygen uptake curves appeared problematic. For 24 substances the values were below 1 mg ThOD/liter. Apparently high Ks values of several tenth mg ThOD/liter for the other 28 substances are explained by retarded mineralization of intermediates due to the relatively high initial test concentrations of up to 300 mg ThOD/liter. At lower test concentrations (down to 15 mg ThOD/liter) the measured Ks values for these substances were also typically below or around 1 mg ThOD/liter. Based on the above-mentioned maximum growth rates and the low values for Ks, the predicted steady-state residual concentration of these substances in the effluent of completely mixed sewage treatment plants is between 10 and 100 micrograms/liter. This level is in accordance with the range of measured values taken from literature. Based on this research and additional data from literature, the separation between readily and inherently degradable can be made at the critical values of mumax approximately 1.5/day and Ks approximately 1 mg ThOD/liter. PMID- 8727515 TI - Effects of lindane on the planktonic community in freshwater microcosms. AB - Effects of lindane on natural planktonic communities were investigated in aquatic indoor microcosms. Lindane was dosed to eight 300-liter microcosms for 2 weeks, and effects and recovery processes were monitored over 12 weeks. Mean measured water concentrations of lindane during the exposure period were 3, 6, 13, 27, 55, 102, 230, and 455 micrograms/liter. Zooplankton was severely affected by lindane. The most sensitive organisms were nauplii of copepodes which were affected at all treatment levels > or = 6 micrograms/liter during exposure (Days 2-14). Recovery of nauplii abundance was concentration-dependent at concentrations < or = 55 micrograms/liter lindane, whereas at concentrations > or = 102 micrograms/liter recovery was negligible. Cladocera abundance decreased during exposure to > or = 102 micrograms/liter lindane and did not recover to original levels until the end of the study. For phytoplankton, decreased population densities of Chlorophyceae and microalgae ( < 5 microns) were observed at lindane concentrations < or = 55 micrograms/liter, where zooplankton was only temporarily affected. At higher lindane, concentrations ( > or = 102 micrograms/liter) effects on phytoplankton were not clear, as also increases in abundances were observed, probably as a consequence of minimized grazing pressure, due to high zooplankton mortality. Comparison of these results with data obtained in complex outdoor systems demonstrates the practicability and sensitivity of indoor microcosms and emphasizes the importance of long-term testing and assessment of recovery processes for prediction of environmental effects. PMID- 8727516 TI - PAH content and mutagenicity of marine sediments from the Venice lagoon. AB - Sediments from the Venice lagoon, a polluted coastal environment in northeastern Italy, were assayed for mutagenicity and content of several toxic microcontaminants, which included selected polycyclic aromatic hydrocarbons (PAHs); the latter are specifically dealt with in this paper. Samples were collected at three lagoon sites with reasonably distinct environmental features- urban, industrial, or agricultural--and at two others considered to be under mixed pollution influences; a sixth sample was obtained from an open sea area to act as background control. The organic matter (EOM) associated with the mineral substrata was extracted; after cleanup, analyte determination was carried out by HRGC-LRMS(SIM) using isotopically labeled compounds as internal standards. Cumulative levels of the selected PAHs were found to be in the range of 0.065 to 0.46 micrograms/g of dry matrix at five sites; a much higher concentration (48 micrograms/g) was detected in the sample from the urban environment. The remarkable PAH level increase at this site was mostly accounted for by the concurrent, apparent increase of EOM contamination as PAH concentration was seen to reach 32 micrograms/mg of EOM from < 1 microgram/mg at the five remaining sites. Mutagenicity assays with Salmonella typhimurium strains TA98 and TA100 of marine sediment organic extracts also highlighted a distinct activity in the sample from the urban site. Further fractionation and analysis of this extract pointed to PAHs as the main mutagenic component present in the sediment matrix, possibly accounting for up to approximately 70-80% of the entire mutagenic potential detected. PMID- 8727517 TI - Determination of cadmium-metallothioneins in zebra mussels exposed to subchronic concentrations of Cd2+. AB - Metallothioneins have been detected and investigated in zebra mussels (Dreissena polymorpha) using high-performance liquid chromatography (size exclusion) coupled with microatomization-AAS or inductively coupled plasma mass spectrometry. The mussels were exposed to 0.2, 2, and 20 micrograms/liter Cd2+ (as CdCl2) for 1 month under controlled temperature and dietary conditions. Elevated (relative to control) concentrations of tissue Cd2+ were detected in all specimens exposed to 2 micrograms Cd/liter and more than 50% of the specimens exposed to 0.2 micrograms Cd/liter, demonstrating that Dreissena cannot regulate Cd2+ at trace exposure concentrations. In most specimens, at least 85% of the measured Cd2+ was bound to metallothioneins. After reduction and exposure to excess Cd2+, the metallothionein fraction of all extracts adsorbed similar quantities of Cd2+, indicating that the physiological concentration of metallothionein in the exposed specimens remained similar to the basal concentration in the control specimens. Thus, a short-term exposure to environmentally relevant concentrations of Cd2+ did not produce a genetic induction of metallothionein biosynthesis as generally observed in specimens exposed to higher concentrations of d10 metals. PMID- 8727518 TI - Metabolites of chlorinated syringaldehydes in fish bile as biomarkers of exposure to bleached eucalypt pulp effluents. AB - Metabolites of chlorinated phenolic compounds in fish bile have been found to be sensitive biomarkers of bleached pulp mill effluent exposure. Chlorinated syringaldehydes are largely unstudied chlorophenolics found in bleached hardwood effluent. Sand flathead (Platycephalus bassensis), Australian marine fish, were exposed to 100% chlorine dioxide-bleached eucalypt pulp effluent at concentrations of 0.5, 2, and 8% (v/v) for 4 days. Metabolites of 2 chlorosyringaldehyde (2-CSA), the predominant chlorophenolic in this effluent, were measured in the bile. The major metabolite was the conjugate of 2-chloro-4 hydroxy-3,5-dimethoxy-benzylalcohol (2-CB-OH), the reduced product of 2-CSA. 2-CB OH was found in all fish exposed to diluted effluent and was concentrated in the bile over 1000 times above 2-CSA levels in the effluent. A separate experiment examined the metabolic fate of 2,6-dichlorosyringaldehyde (2,6-DCSA), which is one of the major chlorophenolics in chlorine-bleached eucalypt pulp effluent. Sand flathead were exposed to 2,6-DCSA by intraperitoneal injection at 15 mg/kg or through the water to 0.5, 2, or 8 micrograms/liter for 4 days. Analysis of the bile revealed the major metabolite of 2,6-DCSA to be the conjugate of 2,6 dichloro-4-hydroxy-3,5-dimethoxybenzylalcohol, which was found in all exposed fish and was concentrated in the bile over 20,000 times above 2,6-DCSA exposure levels. Results reveal that the analysis of metabolites of chlorinated syringaldehydes in fish bile can provide a biomarker of bleached hardwood effluent exposure that is sensitive to low levels of exposure, specific to certain bleaching sequences, and correlates well with exposure concentrations. PMID- 8727519 TI - Preadapted inocula for limiting the risk of errors in biodegradability tests. AB - Reducing the time for biodegradability tests to 28 days poses a problem when the inoculum contains few biodegraders, as a biodegradable xenobiotic must give a positive result within this time. The influence of initial concentration (X0, number of cells liter-1) on the lag time (hours) of para-nitrophenol biodegradability tests was examined using different concentrations of adapted Pseudomonas putida with para-nitrophenol as the sole carbon and energy source. Lag time decreased as bacterial density increased according to the expression y = 63.5 - 5.7(log10X0). The addition of river water to the P. putida concentrations shortened the lag time-bacterial density relationship and lag time filled the expression y = 52.4 - 5.1(log10X0). The addition of river water also increased the rate of para-nitrophenol biodegradation from 1.29 mgC (liters x hr)-1 to 2.11 mgC (liters x hr)-1. An examination of the effect of the initial adapted P. putida density, expressed as total cell, cultivable bacteria, or biodegraders, was also made on the para-nitrophenol biodegradability test outcome. The model related cell density and the probability of test response give very similar k constants (kT = 0.56 x 10(-3) liter total cells-1; kv = 0.11 x 10(-3) liter CFU 1, kMPN = 0.16 x 10(-3) liter MPN-1). Comparisons with nonadapted natural mixed culture (activated sludge, river water) indicate that the biodegradability test responses were the same as with adapted cells when the nonadapted cell concentrations were at least 5 x 10(10) total cells liter-1. As this high cell concentration led to carbon contamination, adapting mixed inocula before the test to increase the number of biodegraders appears to be the best solution. Before biodegradability tests, cell density can be adjusted using techniques which are not specific to biodegraders, and only 10(5) total adapted cells liter-1 are needed for a 99.9% chance of a positive response in para-nitrophenol biodegradability tests. PMID- 8727520 TI - Lack of dietary calcium effect on chlordecone increased white blood cell count, total iron, and iron-binding capacity in serum of rat. AB - Male Sprague-Dawley rats were treated with 0, 1, 10, 50, and 100 ppm of chlordecone (Cd) mixed in calcium-sufficient (Ca-S) or calcium-deficient (Ca-D) diet for 15 days. The control rats fed with Ca-D diet exhibited a significant increase in white blood cell (WBC) counts compared to the rats fed with Ca-S diet. Dietary calcium (Ca), however, did not elicit any significant effect on total iron content and iron-binding capacity (transferrin) of control rats, whereas Cd at higher concentrations significantly increased WBC counts, total iron, and iron-binding capacity in serum of both Ca-S and Ca-D rats. The data suggest that dietary Ca did not alter Cd-increased WBC count, total iron, and iron-binding capacity in serum of rat. PMID- 8727521 TI - Functional effects of the bacterial insecticide Bacillus thuringiensis var. kurstaki on aquatic microbial communities. AB - Epilithic microbial communities were colonized on leaf disks and exposed to commercial preparations of Bacillus thuringiensis var. kurstaki (Btk) in aquatic microcosms. Responses in terms of microbial respiration, bacterial cell density, protozoan density, and microbial decomposition activity were measured. Test concentrations for treatments with Dipel 64AF and Dipel 8AF in microcosms were the expected environmental concentration (EEC) of 20 IU/ml, 100x the EEC, and 1000x the EEC. Bacterial cell density in the biofilm of leaf disks was significantly increased at concentrations as low as the EEC. There were no concomitant alterations in protozoan density. Microbial respiration was significantly increased, and decomposition activity was significantly decreased, but only at the artificially high concentration of 1000x the EEC. This effect was attributed to the spore-crystal component rather than formulation ingredients. Microbial decomposition of leaf material was also determined in outdoor stream channels treated at concentrations ranging from the EEC to 100x the EEC. Although there tended to be reduced decomposition activity in treated channels, there were no significant differences in mass loss of leaf material between treated and control channels. Various regression, classification, and ordination procedures were applied to the experimental data, and none indicated significant treatment effects. These results from laboratory and controlled field experiments indicate that contamination of watercourses with Btk is unlikely to result in significant adverse effects on microbial community function in terms of detrital decomposition. PMID- 8727522 TI - Isolation of a metal-binding protein from ovaries of dab (Limanda limanda L.) distinct from metallothionein: effect of cadmium exposure. AB - Metal-binding proteins were isolated from ovaries of dab (Limanda limanda L.) caught in the North Sea. Separation of the heat-treated cytosolic fraction of dab ovaries by gel-filtration chromatography revealed the presence of a major zinc binding protein with a low-molecular-weight of 10,000-12,000 Da, similar to mammalian metallothionein (MT). The metal-binding protein was analyzed by anion exchange chromatography and by reversedphase high-performance liquid chromatography. The metal-binding protein from dab ovaries consists of one major isoform. Zinc could be displaced from the isoform by a saturation of the extract with cadmium, leading to altered retention of the protein at the RP column. In a laboratory experiment, dabs were treated intraperitoneally with cadmium (0.5 and 1.5 mg/kg body wt). In contrast to hepatic dab MT, the metal-binding protein in ovaries was not inducable by cadmium; however, dose-dependent amounts of cadmium were bound to the ovarian protein after the metal exposure. The amino acid composition was different from that of mammalian MT. The results suggest that the metal-binding protein in dab ovaries is possibly involved in ovary development and can be affected by cadmium exposure. PMID- 8727523 TI - Capture and confinement stress in white sucker exposed to bleached kraft pulp mill effluent. AB - This study evaluates the effects of handling and confinement stress and a 3-day recovery period on a number of biochemical parameters used to monitor exposure of fish to bleached kraft mill effluent (BKME). Plasma was collected at four times of the day from male and female white sucker subjected to four levels of handling stress during their spawning migration at a BKME and a reference site. Indicators of a general response to stress (plasma cortisol, glucose, lactate, and total protein) and of reproductive fitness (plasma testosterone, 11-ketotestosterone, and 17 beta-estradiol) were measured. With the exception of 17 beta-estradiol in females, all of the parameters measured varied with time of day and stress level. The general stress indicators ranged from being highly to marginally responsive to handling and confinement stress and were inconsistent in terms of indicating a site difference. The reproductive steroids were moderately responsive to stress and gave the most consistent site difference with testosterone and 11 ketotestosterone levels reduced in BKME-exposed fish under low levels of stress. There was limited evidence for recovery from capture, handling, and transport in the spring for plasma cortisol, lactate, and 17 beta-estradiol at the BKME site, plasma glucose at the reference site, and plasma testosterone at both sites. Fish were also more responsive to an additional acute stress on Day 1 and Day 3 of recovery compared to that on Day 0. This study emphasizes the need for standardized methods in field collections and sampling with the least amount of stress possible, and suggests that holding white sucker for 1 or 3 days does not allow them to recover from the stress of capture. PMID- 8727524 TI - Alveolar epithelial clearance of protein. AB - Substantial progress has been made in understanding the rate, the pathways, and the mechanisms regulating alveolar protein removal from the uninjured lung. Whole animal studies and cellular studies have demonstrated that the majority of alveolar epithelial protein clearance occurs by passive nondegradative diffusional pathways. Some evidence, however, has been recently presented that alveolar epithelial cells express an albumin-binding receptor as well as a polymeric immunoglobulin receptor, both of which might be important for alveolar epithelial clearance of protein. However, the contribution of these receptors requires further studies. Little is known about alveolar clearance of protein during pathological conditions; further studies are required to determine the roles of the different cell types in the lung for removal of protein from the alveolar spaces of the lung. Alveolar macrophages are likely to play an important role in the degradation and removal of insoluble protein from the distal air spaces after acute lung injury. In conclusion, the present data suggest that most proteins and peptides deposited on the epithelial surfaces in the distal air spaces are cleared as intact molecules, predominantly via paracellular routes. The contribution of pinocytic processes appear to be of minor importance for translocation of bulk quantities of proteins or peptides across the alveolar epithelium. PMID- 8727525 TI - "Fractal dynamics of human gait: stability of long-range correlations in stride interval fluctuations". PMID- 8727526 TI - Fractal dynamics of human gait: stability of long-range correlations in stride interval fluctuations. AB - Fractal dynamics were recently detected in the apparently "noisy" variations in the stride interval of human walking. Dynamical analysis of these step-to-step fluctuations revealed a self-similar pattern: fluctuations at one time scale are statistically similar to those at multiple other time scales, at least over hundreds of steps, while healthy subjects walk at their normal rate. To study the stability of this fractal property, we analyzed data obtained from healthy subjects who walked for 1 h at their usual, slow, and fast paces. The stride interval fluctuations exhibited long-range correlations with power-law decay for up to 1,000 strides at all 3 walking rates. In contrast, during metronomically paced walking, these long-range correlations disappeared; variations in the stride interval were random (uncorrelated) and nonfractal. The long-range correlations observed during spontaneous walking were not affected by removal of drifts in the time series. Thus the fractal dynamics of spontaneous stride interval are normally quite robust and intrinsic to the locomotor system. Furthermore, this fractal property of neural output may be related to the higher nervous centers responsible for the control of walking rhythm. PMID- 8727527 TI - Specific ventilation distribution in microgravity. AB - We studied the contribution of inter- and intraregional inhomogeneities of specific ventilation (delta V/Vo) from the rebreathing inert gas trace in microgravity and on Earth. The rebreathing tests were carried out by four astronauts before, during, and after the 10-day Spacelab D-2 mission. Starting from functional residual capacity, the rebreathing maneuver consisted of eight reinspirations from a bag filled with 1.8-2.2 liters of test gas mixtures containing approximately 5% argon. The rate of argon equilibration in the rebreathing bag, termed RBeq, was quantified by determining the logarithm of the actual minus the equilibrated argon concentrations normalized to the inspired minus the equilibrated argon concentrations. A compartmental model of the lung (S. Verbanck and M. Paiva. J. Appl. Physiol. 76: 445-454, 1994) was used to validate the method for determining RBeq and to simulate the influence of intra- and interregional delta V/Vo inhomogeneities on the RBeq curve. The comparison between the experimental Earth-based and microgravity RBeq curves and model simulations shows that gravity-independent delta V/Vo inhomogeneity is at least as large as gravity-dependent delta V/Vo inhomogeneity. PMID- 8727528 TI - Effects of genioglossal response to negative airway pressure on upper airway collapsibility during sleep. AB - Continuous negative airway pressure (CNAP) trials can be used to measure upper airway (UA) collapsibility. This procedure can be accompanied by an increase in UA muscle activity. The purpose of this study was to evaluate the influence of CNAP-induced increase in genioglossal (GG) activity on UA collapsibility in 10 healthy sleeping men. UA collapsibility was measured on two occasions; each recording was preceded by the administration of a placebo or diazepam (0.15 mg/kg) in a randomized double-blind crossover design. In seven subjects, the decrease in mask pressure (Pmask) was associated with an increase in mean GG electromyographic (EMG) activity during the placebo trial, with a significant negative relationship between these two variables. This relationship was still observed with diazepam. In six subjects, the slope of the relationship between mean EMG and Pmask was less negative with diazepam. This was associated with an increase in critical pressure (Pcrit). With the placebo, a positive relationship was found between Pcrit and the slope of the mean EMG/Pmask relationship. We conclude that Pcrit is influenced by the GG response to the decrease in Pmask during CNAP. PMID- 8727529 TI - Ventilatory dynamics during transient arousal from NREM sleep: implications for respiratory control stability. AB - The polysomnographic and ventilatory patterns of nine normal adults were measured during non-rapid-eye-movement (NREM) stage 2 sleep before and after repeated administrations of a tone (40-72 dB) lasting 5 s. The ventilatory response to arousal (VRA) was determined in data sections showing electrocortical arousal following the start of the tone. Mean inspiratory flow and tidal volume increased significantly above control levels in the first seven breaths after the start of arousal, with peak increases (64.2% > control) occurring on the second breath. Breath-to-breath occlusion pressure 100 ms after the start of inspiration showed significant increases only on the second and third postarousal breaths, whereas upper airway resistance declined immediately and remained below control for > or = 7 consecutive breaths. These results suggest that the first breath and latter portion of the VRA are determined more by upper airway dynamics than by changes in the neural drive to breathe. Computer model simulations comparing different VRA time courses show that sustained periodic apnea is more likely to occur when the fall in the postarousal increase in ventilation is more abrupt. PMID- 8727530 TI - Anatomic dead space in infants and children. AB - In adults, anatomic dead space is 2.2 ml/kg. Because of the relatively large head size of infants and children, we hypothesized that extrathoracic and, therefore, total dead space would be relatively larger in pediatric subjects. Extrathoracic dead space was measured by a "water displacement" technique in 40 patients aged 7 days to 14.2 yr who were intubated with cuffed endotracheal tubes. Intrathoracic dead space was measured by continuous analysis of end-tidal and mixed-expired PCO2 and minute ventilation in 10 patients, aged 18 days to 14.7 yr. Extrathoracic dead space per kilogram decreased exponentially with increasing age, ranging from 2.3 ml/kg in early infancy to 0.8 ml/kg in children older than 6 yr. Mean intrathoracic anatomic dead space was 1.03 ml/kg and was not related to age. The following relationship between total anatomic dead space (DStotal; in ml/kg) and age (in yr) is derived: DStotal = 3.28 - 0.56 [ln(1 + Age)], with r = 0.95 and P = 0.0001. Anatomic dead space is age dependent and is > 3 ml/kg in early infancy. PMID- 8727531 TI - On the mechanism of the mediolateral gradient of parasternal activation. AB - Recent studies have shown that in spontaneously breathing dogs the parasternal intercostals are activated according to a mediolateral gradient. To assess the mechanism of this regionalization of activity, we assessed the pattern of activation of these muscles after section of the dorsal roots and examined the topographic distribution of the muscle fiber types from the sternum to the chondrocostal junctions. The pattern of parasternal activity after dorsal rhizotomy was similar in all respects to that previously observed in intact animals. Thus activity in the medial parasternal bundles at the onset of inspiration frequently preceded activity in the middle bundles, and no activity was recorded from the lateral bundles. The amount of medial activity, when expressed as a percentage of the activity recorded during supramaximal tetanic stimulation of the internal intercostal nerve (maximal activity), was also consistently greater than the amount of middle activity (52.6 +/- 4.6 vs. 23.1 +/ 2.6% maximal activity; P < 0.001). Furthermore, the medial, middle, and lateral parasternal bundles had a higher proportion of slow-twitch oxidative fibers than of fast-twitch oxidative-glycolytic fibers; no topographic difference in fiber type distribution was observed. We conclude, therefore, that the mediolateral gradient of parasternal activity is probably due to the unequal distribution of central inputs throughout the pool of alpha-motoneurons. PMID- 8727532 TI - Expiratory muscle endurance performance after exhaustive submaximal exercise. AB - The aim of our study was to describe the endurance capacity of the expiratory muscles and to determine whether it is altered after exhaustive cycling exercise. Subjects performed repeated maximal expiratory efforts against a closed breathing valve, with and without prior exercise performed at a work rate that elicited 75% of the maximum ventilation rate. Each expiratory effort lasted 6 s, was separated by 10 s of rest, and was initiated from the end-expiratory lung volume. Endurance performance was assessed by measuring the decline in area under the pressure*time curve over 39 contractions. Prior exhaustive exercise attenuated the ability to generate and sustain maximal expiratory pressure (P = 0.013) and resulted in significant declines in the integrated electromyogram of the rectus abdominis (P = 0.005) and external oblique (P = 0.036) abdominal muscles. Each subject also performed a handgrip endurance task before and after exhaustive exercise on a separate day. Prior exercise had no effect on handgrip endurance performance, suggesting that the decline in expiratory muscle performance after exercise was not the result of reduced motivation. We conclude that the ability to maximally activate the abdominal expiratory muscles and to generate maximum expiratory pressure is impaired after exhaustive exercise. Declines in the surface integrated electromyogram despite maximal effort is consistent with findings in limb muscles and is thought to be due to a slowing of motoneuron firing rates or to neuromuscular transmission failure. PMID- 8727533 TI - Influence of mechanical loading on myosin heavy-chain protein and mRNA isoform expression. AB - The overall objective of the studies reported herein was to examine the effects of high-resistance training on myosin heavy-chain (MHC) protein and mRNA isoform expression. The findings from these studies can be summarized as follows: 1) there was a substantial increase in the fast type IIX MHC protein isoform content of the trained red and white medial gastrocnemius muscles, but this did not occur until after the eighth training session (i.e., 16 days); 2) single-fiber analyses demonstrated that many so-called fast type IIB fibers contained small amounts of the fast type IIX MHC protein isoform and that the high-resistance training program altered the bias of fast type IIB-type IIX MHC protein isoform distribution in these fibers but did not increase the number of fibers that could be categorized as exclusively fast type IIX fibers; 3) the high-resistance training program produced a rapid (i.e., after two training sessions) elevation in the fast type IIX MHC mRNA isoform and a corresponding repression of the fast type IIB MHC mRNA isoform; and 4) the dose-response study revealed that as few as 10 contractions (40 s) per training session were capable of elevating the expression of the fast type IIX MHC mRNA isoform by approximately 250%. These collective findings demonstrate that high-resistance training is a powerful modulator of MHC protein isoforms and that pretranslational mechanisms are very sensitive to even small amounts of high-resistance training. PMID- 8727534 TI - Lung water is increased in regions of higher neutrophil retention after acute bead embolization. AB - Previous reports have shown that neutrophils are retained in the lung after acute embolization and that these neutrophils play an important role in the subsequent formation of permeability pulmonary edema. The present study was designed to test the hypothesis that acute embolic injury results in microvascular damage in lung regions with the greater retention of neutrophils. Seventeen pigs (20 +/- 2 kg) were embolized by injecting polystyrene beads (250 microns; labeled with 131I) into the right atrium over 5 min. Five pigs, which received no embolic beads, served as controls. Neutrophils (89 +/- 5% pure), isolated on Ficoll-Histopaque gradient, were radiolabeled with 111In-oxine. Twenty minutes after embolization, the radiolabeled neutrophils were injected into the right atrium along with 85Sr labeled microspheres to mark the initial neutrophil distribution within the lung as well as the regional pulmonary blood flow at the time of their delivery. The animals were killed 50 min after embolization, and the lungs were removed, frozen over liquid nitrogen, and cut into 60 samples. The data show that after embolization regional neutrophil retention was inversely related to the regional blood flow but was not affected by the embolic load in the same region. Regional extravascular lung water was increased in regions of higher neutrophil retention, but the regions with increased edema did not receive a greater embolic load. These results show that microvascular injury occurs in the lung regions with the greatest neutrophil retention and that this increased retention of neutrophils is unrelated to the extent of embolization. PMID- 8727535 TI - Control of nasal dilator muscle activities during exercise: role of nasopharyngeal afferents. AB - Our primary aim was to determine whether reducing the activity of nasal airway receptors would influence drive to the nasal dilator muscles (NDMs) during exercise. We used lidocaine (2%) or nasal splints to diminish afferent airway receptor activity and measured the electromyogram (EMG) activity of the NDMs during incremental bicycle exercise in subjects who breathed nasally. NDM EMG activities increased as a function of exercise intensity but were not changed by lidocaine and were only slightly reduced by splinting. Similarly, neither intervention altered the normal decrease in NDM EMG activity associated with reductions in airway resistance evoked by He-O2 breathing. We also compared the NDM EMG response to exercise with that evoked by CO2 rebreathing at rest to determine whether the nature of the ventilatory stimulus influences drive to the NDMs; comparisons were made at constant levels of nasal inspired ventilation and, therefore, constant total ventilatory output. The increase in EMG activity was much higher during exercise compared with hyperoxic hypercapnia. In conclusion, 1) desensitizing the nasal airway does not alter NDM activity significantly during exercise and 2) exercise results in much greater increases in NDM activity compared with hypercapnia, indicating that different ventilatory stimuli can evoke more or less activation of upper airway motoneurons, even when comparisons are made at constant levels of total ventilatory output. PMID- 8727536 TI - Negative pressure-induced deformation of the upper airway causes central apnea in awake and sleeping dogs. AB - We investigated the effects of negative pressure (NP) in the isolated upper airway (UA) in three unanesthetized dogs. The UA was isolated, and the dogs breathed through an endotracheal tube while wearing a fitted fiberglass snout mask. NP (-2 to -32 cmH2O) was applied in a square wave below the larynx or at the snout at end expiration and was held until inspiratory effort during wakefulness, non-rapid-eye-movement (NREM) sleep, and rapid-eye-movement (REM) sleep. During all states of consciousness, NP applied to the UA prolonged expiratory time (TE) 1) below a threshold of -8 to -10 cmH2O, which coincided with closure of the oro- and/or velopharynx; and 2) in a progressive fashion at more negative pressures than threshold, up to a mean apneic length of 324% of the control value (or 13.9 s) at -30 cmH2O. TE prolongation was less during REM sleep at a given NP (P < 0.05). Augmented tonic genioglossal electromyographic activity also occurred with the applied NP during wakefulness and NREM sleep but not with REM sleep. NP (-20 to -32 cmH2O) applied as a brief pulse (300-500 ms) during NREM sleep caused transient airway occlusion, terminated the breath during inspiration, and prolonged TE when applied at end expiration. Central apneas always persisted beyond the termination of the UA closure. TE prolongation in response to NP persisted in the presence of a topical anesthetic nebulized through the UA sufficient to abolish the laryngeal gag reflexes. We conclude that UA closure and deformation will cause significant TE prolongation during all states of consciousness and activation of the genioglossus muscle during wakefulness and NREM sleep but not during REM sleep. PMID- 8727537 TI - Role of glucose transport in glycogen supercompensation in reweighted rat skeletal muscle. AB - Hindlimb weight bearing after a 3-day period of hindlimb suspension (reweighting) of juvenile rats results in a marked transient elevation in soleus glycogen concentration that cannot be explained on the basis of the activities of glycogen synthase and phosphorylase. We have hypothesized that enhanced glucose transport activity could underlie this response. We directly tested this hypothesis by assessing the response of insulin-dependent and insulin-independent glucose transport activity (in vitro 2-[1,2-3H]deoxy-D-glucose uptake) as well as glucose transporter (GLUT-4) protein levels during a 48-h reweighting period. After a net glycogen loss (from 29 +/- 2 to 16 +/- 1 nmol/mg muscle; P < 0.05) during the first 2 h of reweighting, glycogen accumulated at an average rate of 1.4 nmol.mg 1.h-1 up to 18 h, reaching an apex of 38 +/- 1 nmol/mg. During this same reweighting period, insulin-independent, but not insulin-dependent, glucose transport activity was significantly enhanced (P < 0.05 vs. weight-bearing control values) and was associated with an elevated level of GLUT-4 protein and the specific activity of total hexokinase. The specific activity of citrate synthase was also increased. By 24 h of reweighting, although insulin-independent glucose transport activity and GLUT-4 protein remained elevated, glycogen accumulation had ceased, likely due to enhanced phosphorylase activity at this time point. These results are consistent with the interpretation that the glycogen supercompensation seen during reweighting of the rat soleus may be regulated in part by an enhanced glucose flux arising from an increase in insulin independent glucose transport activity and hexokinase activity. PMID- 8727538 TI - In vitro human masseter muscle hypersensitivity: a possible explanation for increase in masseter tone. AB - To determine whether a difference in fiber-type caffeine and Ca2+ sensitivities exists between human masseter and vastus lateralis skeletal muscle, we compared the fiber-type caffeine sensitivities in chemically skinned muscle fibers from 13 masseter and 18 vastus lateralis muscles. Caffeine sensitivity was defined as the threshold concentration inducing > 10% of the maximal tension obtained after the fiber was loaded with a 1.6 x 10(-2) mM Ca2+ solution for 30 s. Significant difference in the mean caffeine sensitivity was found between type I masseter fibers [2.57 +/- 1.32 (SD) mM] vs. type I (6.02 +/- 1.74 mM) and type II vastus lateralis fibers (11.25 +/- 3.13 mM). Maximal Ca(2+)-activated force per cross sectional area was significantly different between masseter and vastus lateralis fibers. However, the Ca2+ concentration corresponding to half-maximal tension (pCa50) was not significantly different between type I masseter (pCa50 5.9 +/- 0.02) and type I vastus lateralis muscle (pCa50 6.01 +/- 0.08). These results suggest that the increase in caffeine sensitivity of masseter muscle reflects the presence of a low reactivity threshold of the sarcoplasmic reticulum. PMID- 8727539 TI - Smoking increases conversion of lactate to glucose during submaximal exercise. AB - We examined the hypotheses that 1) smoking acutely before exercise (AS) results in a higher rate of lactate production during exercise compared with chronic smoking with preexercise abstinence (CS) and 2) smokers have a higher rate of lactate conversion to glucose during exercise compared with nonsmokers (NS). To test our hypotheses, seven male smokers and seven nonsmokers were studied by using a primed continuous infusion of [3-13C]lactate during 90 min of rest and 60 min of exercise on a cycle ergometer at 50% peak O2 consumption; smokers were studied twice: once after an overnight smoking abstinence and once after smoking three cigarettes before exercise. The rates of lactate appearance and conversion to glucose were increased markedly with exercise compared with rest in all groups (P < 0.05); the rate of lactate appearance for AS was significantly greater (7.87 +/- 0.77 mg.kg-1.min-1) than for both CS (4.64 +/- 0.33 mg.kg-1.min-1) and NS (5.57 +/- 0.60 mg.kg-1.min-1) (P < 0.05). The rate of lactate conversion to glucose was similar between CS and AS (6.49 +/- 1.82 and 6.30 +/- 1.69 mg.kg 1.min-1, respectively) during exercise; NS had a significantly lower rate (3.31 +/- 0.90 mg.kg-1.min-1) compared with CS and AS (P < 0.05). In summary, acute smoking increases lactate flux during exercise; in addition, smokers have a higher rate of lactate to glucose conversion during exercise compared with nonsmokers, which may indicate an increased glucose dependency. PMID- 8727540 TI - Alterations in surfactant protein A after acute exposure to ozone. AB - The surfactant layer covering the gas-exchange region of the lung serves as the initial site of interaction with inhaled oxidant gases. Among the endogenous compounds potentially vulnerable to oxidative injury are surfactant proteins. This study focused on the effect of ozone on surfactant protein A (SP-A) function, content, and gene expression. To determine the time course of response to ozone, guinea pigs were exposed to 0.2-0.8 parts/million (ppm) ozone for 6 h and were killed up to 120 h postexposure. To determine the effect of repeated exposure, animals were exposed to 0.8 ppm ozone for 6 h/day and were killed on days 3 and 5. A significant increase in surfactant's ability to modulate the respiratory burst induced by phorbol 12-myristate 13-acetate in naive macrophages was observed at 24 h after a single 0.8 ppm ozone exposure. Because neutralizing antibodies to SP-A blunted this stimulatory effect, we hypothesized that ozone enhanced the modulatory role of SP-A in macrophage function. This alteration in function was accompanied by an influx of inflammatory cells and only marginal changes in SP-A levels as determined by an enzyme-linked immunosorbent assay. No significant changes in steady-state levels of SP-A mRNA were observed after single or repeated exposure to ozone. Thus the inflammation that accompanies in vivo ozone exposure may result in a change in the structure and thus functional role of SP-A in modulating macrophage activity. PMID- 8727541 TI - Pulmonary capillary diameters and recruitment characteristics in subpleural and interior networks. AB - In vivo microscopic observations of pulmonary capillaries are limited to subpleural networks that are less dense than interior networks. In addition to the density difference, subpleural and interior capillary diameters may differ, although there are conflicting data on this point. We measured the diameters of subpleural and interior capillaries in rats and dogs. Subpleural diameters were 30% larger in rats and 20% larger in dogs. Because diameter and density differences might cause differences in recruitment between subpleural and interior networks, we measured subpleural and interior recruitment by counting the number of red blood cells per 10 microns of alveolar wall in histological cross sections of rapidly frozen rat lungs. Lung inflation pressures of 4, 12, and 25 cmH2O created a wide range of capillary recruitment in different groups of animals. Red blood cell counts for interior and subpleural capillaries moved in parallel and progressively increased as inflation pressures were reduced. These data demonstrate that recruitment in subpleural capillaries accurately reflect recruitment in interior capillaries and validate the use of in vivo microscopic observations of subpleural capillaries to investigate pulmonary capillary recruitment in general. PMID- 8727542 TI - Importance of muscle phosphocreatine during intermittent maximal cycling. AB - To examine the importance of phosphocreatine (PCr) degradation in maintaining power output during maximal intermittent cycling, seven healthy men completed three bouts of isokinetic cycling (30 s, 100 revolutions/min) with 4 min of rest between bouts. After bout 2, blood flow to one leg was occluded by cuffing the thigh (Cuff) during the rest period to prevent PCr resynthesis while the circulation to the other leg was intact (Cont). The cuff was then removed and bout 3 completed. Muscle biopsies were sampled from the vastus lateralis of both legs just before and immediately after bout 3. Total work produced by the Cuff and Cont legs was similar during bouts 1 (9.3 +/- 0.5 and 9.6 +/- 0.5 kJ, respectively) and 2 (8.1 +/- 0.4 and 8.3 +/- kJ, respectively). Cuffing prevented the resynthesis of PCr because pre-bout 3 contents were 20.7 +/- 8.4 and 63.0 +/- 3.3 mmol/kg dry muscle in the Cuff and Cont legs, respectively. Cuffing also resulted in significantly higher muscle levels of lactate, H+ concentration (287 +/- 26 vs. 217 +/- 15 nM), ADP, AMP, and acetyl-CoA before bout 3 but had no effect on other glycolytic intermediates, ATP, or acetylcarnitine. Total work in bout 3 was significantly reduced by 15% in the Cuff leg (5.8 +/- 0.4 vs. 6.8 +/- 0.6 kJ). PCr degradation during bout 3 was 3.1 and 47.5 mmol/kg dry muscle in the Cuff and Cont legs, respectively, and lactate accumulation was minimal in both legs. Changes in all other metabolites during bout 3 were not different between legs. The results suggest that PCr contributed approximately 15% of the total ATP provision during the third 30-s bout of maximal isokinetic cycling and that most of the ATP was provided during the initial 15 s. Muscle glycogenolysis contributed minimally to ATP provision (approximately 10-15%) during the third 30 s bout, suggesting that aerobic metabolism becomes the dominant source of ATP during this model of repeated sprinting. PMID- 8727543 TI - Effect of lung inflation and airway muscle tone on airway diameter in vivo. AB - How normal airway dimensions change with lung volume is of great importance in determining flow limitation during the normal forced vital capacity maneuver as well as in the manifestation of obstructive lung disease. The literature presents a confusing picture, with some results suggesting that airway diameter increases linearly with the cube root of lung volume and others showing a highly nonlinear relation. The effect of smooth muscle contraction on lung-airway interdependence is even less well understood. Recent morphological work explicitly assumes that airway basement membrane is nondistensible, although the lung volume at which this maximal airway size is reached is unknown. With smooth muscle contraction, folding of the epithelium and basement membrane accounts for the changes in luminal area. In this study, we measured the effect of lung inflation on relaxed and contracted airway areas by using high-resolution computed tomography at different transpulmonary pressures, each held for 2 min. We found that fully relaxed airways are quite distensible up to a pressure of 5-7 cmH2O (P < 0.001), where they reach a maximal size with no further distension up to an airway pressure of 30 cmH2O (P = 0.49). Thus relaxed airways clearly do not expand isotropically with the lung. With smooth muscle tone, the airways in different animals responded differently to lung inflation, with some animals showing minimal airway dilation up to an airway pressure of 20 cmH2O and others showing airways that were more easily dilated with lung expansion. However, maximal diameter of these moderately constricted airways was not usually achieved even up to an airway pressure of 30 cmH2O. Thus a transient deep inspiration in vivo would be expected to have only a small effect on contracted airways. PMID- 8727544 TI - Acoustic rhinometry: influence of paranasal sinuses. AB - The influence of the maxillary sinuses in acoustic rhinometry (AR) has not been evaluated, and this is the aim of the present study. We examined six subjects with AR and magnetic resonance imaging (MRI) after nasal decongestion to compare the area-distance relationships determined by the two methods. From the MRI data we obtained copies of the nasal cavities with and without maxillary sinuses, which were made in plastic by a stereolithographic method. AR curves from models without maxillary sinuses differed from AR curves with sinuses included but were in agreement with MRI curves without inclusion of sinuses. A similar difference in AR was seen in two subjects before and after the nasal cavities were flushed with saline to fill up the maxillary sinuses. The measured volume in the first 50 mm of the nasal cavity models was unaffected by the sinuses, but the volume in the first 70 mm corresponding to the length of the nasal cavity septum was increased slightly but significantly (from 10.8 to 11.3 cm3; P = 0.05). The presence of maxillary sinuses increased the volume of the epipharynx (70-100 mm from the nostril) from 12.2 to 21.3 cm3 (P < 0.01), and this increase was not due to the influence from the contralateral nasal cavity. We conclude that the maxillary sinuses may significantly contribute to the acoustically determined areas in the posterior part of the nasal cavity and the epipharynx, especially during decongestion, and may explain a part of the difference between area distance curves obtained by AR and MRI, whereas contribution from the contralateral nasal cavity does not. PMID- 8727545 TI - Inspiratory coactivation of the genioglossus enlarges retroglossal space in laryngectomized humans. AB - To investigate the relationship between the electrical activity of the genioglossus (GG-EMG) and associated tongue movement, seven laryngectomized subjects breathing through a tracheal stoma (without pressure or flow change in the upper airway) were studied in the supine position. Tongue movement, with the use of lateral fluoroscopy, and GG-EMG expressed as a percentage of maximum voluntary genioglossal activation were monitored simultaneously during 1) spontaneous inspiration (SI), 2) resistive loaded inspiration (LI), and 3) rapid inspiration (RI). Tongue position during each maneuver was compared with its position during spontaneous expiration. Peak GG-EMG during the three maneuvers was significantly different from each other (SI: 5.4 +/- 1.6, LI: 11.9 +/- 1.8, and RI: 51.6 +/- 9.4 (SE) %, respectively). Associated forward movement of the posterior aspect of the tongue was minimum during SI; however, significant movement was observed during LI, and this was increased during RI. Significant covariance existed between peak GG-EMG and this movement. Genioglossal coactivation with inspiration enlarges the glossopharyngeal airway, particularly in its caudal part. In subjects with intact upper airways, this activation may protect or enhance upper airway patency in an effort-dependent manner. PMID- 8727546 TI - Effects of ovariectomy and exercise training on muscle GLUT-4 content and glucose metabolism in rats. AB - The present study examined the effects of 6 wk of ovarian endocrine deficiency on skeletal muscle GLUT-4 glucose transporter protein and glucose transport activity in sedentary and endurance-trained rats. Female Wistar rats (10 wk old) underwent bilateral ovariectomy (OVX) or sham surgery followed by a 5-wk swim-training protocol. OVX resulted in no significant changes in glycogen or GLUT-4 glucose transporter concentration in the soleus, epitrochlearis, or flexor digitorum brevis (FDB) muscles or in basal and maximally insulin-stimulated 2-deoxy-D-[1,2 3H]glucose (2-[3H]DG) transport in the soleus or epitrochlearis, suggesting that moderate-duration ovarian hormone deficiency does not significantly impair insulin action in skeletal muscle. In contrast, OVX decreased the maximal activation of 2-[3H]DG transport in the FDB by in vitro electrical stimulation. OVX had no significant effect on the training-induced changes in oxidative enzyme activities, GLUT-4 protein expression, glycogen content, or insulin-stimulated 2 [3H]DG transport in the soleus or epitrochlearis. These findings provide the first evidence that ovarian hormone deficiency decreases contraction-stimulated glucose transport in skeletal muscle. PMID- 8727547 TI - Decreased Ca2+ sensitivity of isometric tension in skinned cardiac myocytes from tail-suspended rats. AB - Tail suspension in rats causes a cephalic shift in blood, resulting in a volume load on the heart similar to that observed during microgravity spaceflight or mild heart failure. The present study determined the influence of increased cardiac hemodynamic load on myofilament isometric tension as a function of Ca2+ concentration in skinned cardiac myocytes of control and 7-day head-down tilt Sprague-Dawley rats. Isometric force of single skinned myocytes was measured by attaching cells with adhesive to a force transducer and piezoelectric translator. A significant decrease in the Ca2+ sensitivity of tension was observed in cardiac myocytes from suspended rats [pCa of half-maximal tension (pCa50) of 5.83 +/- 0.03] compared with control rats (pCa50 of 5.94 +/- 0.03). Maximum tension generation and slope of the tension-pCa relationship were unaffected by head-down tilt. Electrophoretic analysis of myofilament proteins indicates differences in expression of proteins in the 50-60 and 100-120 kDa ranges; immunoblot analysis of tubulin (50 kDa) expression indicates no change in the ratio of beta-tubulin to light chain 1 or tropomyosin. Decreased force-producing ability at a given submaximum Ca2+ concentration in cardiac myocytes from suspended rats suggests a decrease in contractility possibly due to changes in cardiac myofilament protein expression following chronic elevated volume load on the heart. PMID- 8727548 TI - Cardiorespiratory interactions during fixed-pace resistive breathing. AB - We tested the hypothesis that the arterial baroreflex was important in the origin of respiratory sinus arrhythmia (RSA) under conditions of normal and resistive breathing. That is, mechanical effects of breathing [indicated by instantaneous lung volume (ILV)] would directly influence left ventricular stroke volume (LVSV), which in turn would influence systolic arterial blood pressure (SABP), causing variation in R-R interval through the baroreflex. Eight healthy young subjects (four men and four women) were monitored in the supine position while breathing with a fixed frequency (0.2 Hz) and tidal volume for 15 min through each of three resistances (R0, R1, and R2) producing inspiratory (-) and expiratory (+) pressures of +/- 1.6, +/- 5.4, and +/- 16.6 cmH2O, respectively. LVSV was estimated by stroke distance [(SDist); by Doppler ultrasound]. There were no differences across R0, R1, and R2 for the mean values of R-R interval, SDist, or SABP. Cross-spectral analysis showed that, at R0, each value of R-R interval, SDist, and SABP lagged ILV by approximately 80 degrees. At R1 and R2, phase was reduced from ILV to SDist and R-R interval, and the transfer magnitude for SDist (R2 only), SABP, and R-R interval increased. The transfer magnitude from SDist to SABP significantly increased as a function of resistance breathing, whereas that from SABP to R-R interval significantly decreased. There were no changes in phase relationships from SDist to SABP to R-R interval. Thus the magnitude of RSA (ILV to R-R interval) was increased, but the transfer through the arterial baroreflex (SABP to R-R interval) was reduced. Although factors other than the arterial baroreflex are probably involved in the genesis of RSA, the constant phase relationship across the levels of breathing resistance among SDist, SABP, and R-R interval suggests an important functional link caused by mechanical effects of breathing. PMID- 8727549 TI - Neurocirculatory consequences of abrupt change in sleep state in humans. AB - The arterial pressure elevations that accompany sleep apneas may be caused by chemoreflex stimulation, negative intrathoracic pressure, and/or arousal. To assess the neurocirculatory effects of arousal alone, we applied graded auditory stimuli during non-rapid-eye-movement (NREM) sleep in eight healthy humans. We measured muscle sympathetic nerve activity (intraneural microelectrodes), electroencephalogram (EEG; C4/A1 and O1/A2), arterial pressure (photoelectric plethysmography), heart rate (electrocardiogram), and stroke volume (impedance cardiography). Auditory stimuli caused abrupt increases in systolic and diastolic pressures (21 +/- 2 and 15 +/- 1 mmHg) and heart rate (11 +/- 2 beats/min). Cardiac output decreased (-10%). Stimuli that produced EEG evidence of arousal evoked one to two large bursts of sympathetic activity (316 +/- 46% of baseline amplitude). Stimuli that did not alter EEG frequency produced smaller but consistent pressor responses even though no sympathetic activation was observed. We conclude that arousal from NREM sleep evokes a pressor response caused by increased peripheral vascular resistance. Increased sympathetic outflow to skeletal muscle may contribute to, but is not required for, this vasoconstriction. The neurocirculatory effects of arousal may augment those caused by asphyxia during episodes of sleep-disordered breathing. PMID- 8727550 TI - Assessment of respiratory pressure-volume nonlinearity in rabbits during mechanical ventilation. AB - The volume dependence of respiratory elastance makes it difficult to recognize actual changes in lung and chest wall elastic properties in artificially ventilated subjects. We have assessed in six anesthetized, tracheotomized, and paralyzed rabbits whether reliable information on the static pressure-volume (PV) curve could be obtained from recordings performed during step variations of the end-expiratory pressure without interrupting mechanical ventilation. Pressure and flow data recorded during 5- and 10-hPa positive-pressure steps were analyzed in the time domain with a nonlinear model featuring a sigmoid PV curve and with a model that, in addition, accounted for tissue viscoelastic properties. The latter fitted the data substantially better. Both models provided reasonably reproducible coefficients, but the PV curves obtained from the 5- and 10-hPa steps were systematically different. When the PV curves were used to predict respiratory effective elastance, the best predictor was the curve derived from the 10-hPa step with the viscoelastic model: unsigned differences averaged 8.6 +/ 11.1, 26.9 +/- 36.4, and 5.5 +/- 5.8% at end-expiratory pressures of 0, 5, and 10 hPa, respectively. This approach provides potentially useful, although not highly accurate, estimates of respiratory effective elastance-volume dependence. PMID- 8727551 TI - Role of viscoelasticity in the tube model of airway reopening. II. Non-Newtonian gels and airway simulation. AB - The influence of viscoelastic gels as lining fluids on the pressure-velocity relationships in an airway tube model (Gaver et al. J. Appl. Physiol. 69: 74-85, 1990) was examined. A flow instability was observed due to the occurrence of a sol-gel transition in the viscoelastic properties under flow conditions. We further report measurements of the viscoelastic properties of airway secretions. Airway secretions are gels under small strains and have a yield stress of 4-7 dyn/cm2. Secretions from the pharyngeal airway show lower elasticity than secretions from the trachea. The airway reopening process is simulated using a Weibel lung geometry by incorporating the constitutive equations from the model gel studies and utilizing the rheological data on airway secretions. In these simulations, a "popping-open" phenomenon arises from a flow instability in airway generations 8-14 when the rheological properties of the lining fluids are assumed to be similar to those of pharyngeal secretions. On the basis of these studies, the elasticity of airway secretions plays an important role in airway reopening. PMID- 8727552 TI - Estradiol effect on anterior crural muscles-tibial bone relationship and susceptibility to injury. AB - The study's objective was to determine whether estradiol (E2) deficiency alters the functional relationship of muscle to bone and causes a differential increase in injury susceptibility. Ovariectomized 6-wk-old mice were administered E2 (40 micrograms. day-1. kg-1; n = 8) or the oil vehicle (n = 8) for 21 days. The anterior crural muscles of the left hindlimb were then stimulated to produce 150 maximal in vivo eccentric contractions. In vitro functional measurements were then made on the extensor digitorum longus (EDL) muscle and tibia from both the exercised and unexercised legs. The maximal isometric torque produced by the anterior crural muscles before the eccentric contraction protocol and the unexercised EDL maximal isometric tetanic force (P(0)) were higher in E2-treated mice by 18 and 14%, respectively (P < or = 0.03). Both ultimate load and stiffness for the unexercised tibia were higher by 16% in E2-treated mice (P < or = 0.03). The muscle-to-bone relationship of these measurements was unaffected by E2 status (P > or = 0.59). No evidence for increased injury susceptibility was found in either tissue from E2-deficient mice. In fact, the decrement in P(0) was only 36.9 +/- 3.8% in exercised EDL muscles from E2-deficient mice compared with 50.6 +/- 4.2% in exercised muscles from E2-treated mice (P = 0.03). Tibia stiffness was 3.9% higher in bones from exercised legs than in bones from unexercised legs (72.64 +/- 2.77 vs. 69.95 +/- 2.66 N/mm; P = 0.05) with ultimate load showing a similar trend (P = 0.07); no effect of E2 status was observed on these differences (P > or = 0.53). In conclusion, the functional relationship of bone to muscle and the susceptibility to injury in bone are not altered by the presence of E2 in ovariectomized mice; however, E2 does increase injury susceptibility in the EDL muscle. PMID- 8727553 TI - Differential effects of salmeterol on lung endothelial and epithelial leakage in sheep. AB - Salmeterol has been shown to prevent the influx of proteins into the air spaces of lungs of guinea pigs given intravenous histamine. To determine whether the salmeterol acts to stabilize the epithelial or endothelial barrier, we ventilated anesthetized sheep with aerosolized salmeterol before infusing histamine intravenously at a rate of 4 micrograms.kg-1.min-1 for 3 h. Changes in endothelial permeability were assessed by measuring the flow of lymph and proteins from the lungs. The influx of proteins into the air spaces was detected by performing single-cycle lavages to measure the concentration of circulating 125I-albumin in the epithelial lining fluid. Intravenous histamine increased the lymph flow to 13.2 +/- 6.8 ml/h compared with the control value of 5.6 +/- 2.8 ml/h (P < 0.05). Histamine also increased the concentration of 125I-albumin in the epithelial lining fluid from 1.8 +/- 0.9 to 8.5 +/- 2.5% of the plasma concentration (P < 0.01) and the postmortem lung water volume from 3.5 +/- 0.5 to 5.0 +/- 0.8 mg/g dry lung wt (P < 0.05). Pretreatment with 2.5 mg of aerosolized salmeterol prevented the influx of proteins into the air spaces and the increase in the postmortem lung water volume but it also increased the lung lymph flow even further to 20.0 +/- 5.6 ml/h (P < 0.05), increased the lymph-to-plasma protein ratio from 0.77 to 0.91, and tripled the increase in alveolar-arterial oxygen gradient caused by histamine alone. Pretreatment with 2.5 mg of intravenous salmeterol had essentially the same effect as salmeterol administered by aerosol. We conclude that salmeterol decreases lung epithelial permeability but increases lung endothelial permeability due to intravenous histamine in sheep. PMID- 8727554 TI - Postnatal lung responses and surfactant function after fetal or maternal corticosteroid treatment. AB - To evaluate the effect of dose and route of administration of betamethasone on subsequent postnatal lung function, pregnant ewes were randomized at 127 days gestation to receive maternal or fetal intramuscular doses of 0.2 or 0.5 mg/kg body wt betamethasone or saline. At delivery 24 h later, preterm lambs were treated with surfactant and ventilated for 4 h. The lambs exposed to 0.5 mg/kg betamethasone by either the maternal or fetal route had higher Po2 values, lung volumes, dynamic compliances, and ventilatory efficiency indexes, as well as lower ventilatory pressure requirements, than did control animals (P < 0.05). There were no consistent improvements in postnatal lung function for the 0.2 mg/kg dose given to the fetus or ewe. However, measurements of radiolabeled protein in the total lung were decreased in all treatment groups (P < 0.01). Surfactant that was recovered from all groups of lambs and fractionated to isolate the large-aggregate fraction improved lung volumes in preterm rabbits to a greater degree than the surfactant used to treat the lambs (P < 0.05). Surfactant recovered from both groups treated with 0.5 mg/kg betamethasone was less sensitive to inactivation by plasma than was surfactant from the 0.2 mg/kg groups or the controls (P < 0.01). Fetal or maternal treatment with 0.5 mg/kg betamethasone improved postnatal lung function and increased the resistance of surfactant to inactivation. PMID- 8727555 TI - Preservation of alveolar epithelial fluid transport mechanisms in rewarmed human lung after severe hypothermia. AB - Although hypothermia abolishes alveolar fluid clearance in the in situ goat lung and in the ex vivo human lung, it is unknown whether alveolar fluid clearance resumes in lungs that are rewarmed after severe hypothermia. An isosmolar albumin solution was instilled into resected human lungs that were rewarmed to 37 degrees C after hypothermia (7 +/- 3 degrees C), and then alveolar fluid clearance was measured by the concentration of albumin in the alveolar fluid sample after 4 h. In control experiments in lungs that had not been cooled and rewarmed, alveolar fluid clearance was 11 +/- 2% over 4 h. In separate experiments, hypothermia completely abolished alveolar fluid clearance. However, alveolar fluid clearance resumed to a normal level of 12 +/- 1% over 4 h in the lungs that were rewarmed after hypothermia. Amiloride decreased alveolar fluid clearance by 47% in the rewarmed lungs. Terbutaline increased alveolar fluid clearance by nearly 300% in 2-h experiments in the rewarmed lungs (P < 0.05). The results of this study indicate that alveolar sodium-channel transport mechanisms are preserved in resected human lungs that are exposed to rewarming after hypothermia. PMID- 8727556 TI - Diffusion of nonelectrolytes in the canine trachea: effect of tight junction. AB - We recently demonstrated through theoretical modeling that the exhaled ethanol (EtOH) profile from humans is consistent with a molecular diffusion coefficient (cm2/s) in the bronchial mucosa (Dti) that is only 8% of the diffusion coefficient in water (Dw; J. Appl. Physiol. 75: 2439-2449, 1993). Because of the small oil-water partition coefficient (lambda o:w) of EtOH (lambda o:w = 0.074), the reduced diffusion coefficient may be due, in part, to the epithelial tight junction in the paracellular pathway. We hypothesized that opening the tight junction would open an aqueous pathway and increase the diffusion coefficient of small (mol wt < 100) hydrophilic compounds. We mounted the mucosa from the membranous canine trachea in an Ussing-type diffusion cell and measured the diffusion coefficient of 2-ethoxyethanol (2-Ethx; lambda o:w = 0.042), EtOH, and methyl ethyl ketone (MEK; lambda o:w = 1.04) in the presence and absence of the epithelial tight junction. The tight junction was opened using a phosphate buffered saline free of Ca2+ and Mg2+ with 0.5 mM ethylene glycol-bis (beta aminoethyl ether)-N,N,N',N'-tetraacetic acid, and its integrity was assessed by measuring the transepithelial electrical resistance. Dti/Dw in the presence of Ca2+ and Mg2+ was 0.39, 0.34, and 0.39 for 2-Ethx, EtOH, and MEK, respectively, and increased 24.6, 11.7, and 1.11% in the absence of Ca2+ and Mg2+. We conclude that the effect of the tight junction on Dti increases with increasing water solubility but can account for only a small portion of the reduced Dti of EtOH as predicted by exhaled profiles. PMID- 8727557 TI - How inhomogeneities and airway walls affect frequency dependence and separation of airway and tissue properties. AB - It has been proposed that during mild-to-moderate bronchoconstriction one can partition airway and tissue properties on the basis of input impedance (Zin) acquired from 0.1 to 5 Hz (K.R. Lutchen, B. Suki, Q. Zhang, F. Petak, B. Daroczy, and Z. Hantos. J. Appl. Physiol. 77: 373-385, 1994). The approach is to apply a homogeneous lung model that contains airway resistance and viscoelastic tissue damping and elastance parameters. The tissue parameters account for the frequency dependence in lung resistance (RL) and elastance (EL). We present an anatomically consistent asymmetrically branching airway model to address two key questions: 1) How will lung inhomogeneities, airway wall shunting, and tissue viscoelasticity contribute to increased frequency dependence and levels of RL and EL during lung constriction? and 2) How much can lung inhomogeneities and airway wall shunting contribute to our assessment of airway, tissue, and overall lung properties derived from Zin? The model incorporates nonrigid airway walls and allows for explicit control over the type and degree of inhomogeneous airway constriction or tissue changes. Our results indicate that, from 0.1 to 5 Hz, airway wall shunting does not become important unless the entire lung periphery experiences significant constriction. Mild-to-moderate inhomogeneous peripheral airway constriction produces a relatively minor additional frequency dependence in RL and EL beyond that due to the tissues alone. With more extreme constriction, however, there is a marked frequency-dependent increase in EL. This phenomenon may render it impossible to distinguish from a single frequency measurement whether an increase in EL during bronchoconstriction is a consequence of a true increase in tissue stiffening or simply a consequence of airway phenomena. Finally, Zin from 0.1 to 5 Hz can be used to provide a reasonable separation of airway and tissue properties for mild-to-moderate homogeneous or inhomogeneous lung constriction. However, during more severe disease, inhomogeneities and/or wall shunting will produce substantial overestimation of tissue damping and hysteretic properties. In fact, the only reliable indicator of a real change in the tissues may be a change in the estimate of tissue elastance that is based on data extending to a sufficiently low frequency. PMID- 8727558 TI - Alteration of human cutaneous afferent discharges as the result of long-lasting vibration. AB - The unitary activities of slowly (15 SAI) and fast-adapting type I (12 FAI) skin mechanoreceptive afferent units innervating the anterior part of the human leg and foot were recorded by using the microneurographic method. The recordings were performed both at rest and on application of cutaneous stimuli of various intensities before and after exposure of the corresponding receptive fields to vibration (0.5 mm peak to peak, 100 pulses/s, 10 min). The results show that 11% of the units tested, which were previously silent, developed a bursting pattern of postvibratory activity, which lasted 12 min on average. This induced resting activity may account for the tingling sensations usually perceived after exposure to vibration. Furthermore, application of vibration to the cutaneous receptive fields impaired the response properties of the corresponding cutaneous fibers much more markedly in the case of the SAI than in the FAI units. More specifically, less than one-half of the FAI fibers tested showed a postvibratory depressed sensitivity to skin stroking applied at various velocities that persisted for only a few minutes, whereas the responses of all the SAI units to suprathreshold maintained skin indentations applied with increasing amplitudes decreased significantly for 20 min. These fairly durable changes in the transductive properties of the mechanoreceptive afferent units probably lead to an impairment of perceptual and sensorimotor processes and consequently may at least partly account for the alterations in sensorimotor performance that have been reported to occur in humans after exposure to vibration. PMID- 8727559 TI - Lung mechanics and pulmonary but not systemic vascular responses to ET-1 are Tx and infusion rate dependent. AB - The role of cyclooxygenase metabolites formation in the systemic and pulmonary vascular and airway responses to different intravenous infusion rates of endothelin-1 (ET-1) was investigated in eight barbiturate-anesthetized mechanically ventilated adult sheep. ET-1 (20, 200, and 400 pmol/kg) was infused into the femoral vein over either 1, 10, or 180 s before and after pretreatment with indomethacin (1.5 mg/kg i.v.). ET-1 infusion produced a dose-dependent systemic vasoconstriction that was similar with all three infusion rates. In contrast, the pulmonary vascular and airways responses to ET-1 were not only dose dependent but also infusion rate dependent so that consistent effects on the pulmonary vasculature and airways were observed only when the peptide was injected over 1 s. At the highest dosage and at the fastest rate of administration, ET-1 produced a fivefold rise in pulmonary vascular resistance, a twofold rise in airway resistance, and a 45% decrease in dynamic pulmonary compliance, whereas no changes were observed when the peptide was injected over 180 s. Plasma levels of 6-ketoprostaglandin F1 alpha (6-keto-PGF1 alpha) increased 20-fold when ET-1 was administered over 1 s but only 5-fold when it was administered over 180 s. Thromboxane B2 (TxB2) increased 5-fold when ET-1 was administered over 1 s and did not increase when ET-1 was given over 180 s. Plasma TxB2 levels were linearly correlated with pulmonary vascular or airway resistance during the bolus ET-1 infusion. Pretreatment with indomethacin completely prevented the ET-1-induced rise in TxB2 and 6-keto-PGF1 alpha and blocked pulmonary vaso- and bronchoconstriction observed, whereas it enhanced systemic vasoconstriction. These results demonstrate that in adult sheep intravenous ET-1 produces pulmonary vaso- and bronchoconstriction that is infusion rate dependent and is associated with the rate-dependent production of thromboxane. In contrast, the increase in systemic vascular tone elicited by ET-1 is not affected by its rate of infusion and does not depend on the secondary generation of cyclooxygenase metabolites. PMID- 8727560 TI - Cardiovascular and ventilatory response to isocapnic hypoxia at sea level and at 5,050 m. AB - To assess the effect of chronic hypoxic conditions on ventilatory, heart rate (HR), and blood pressure (BP) responses to acute progressive isocapnic hypoxia, we studied five healthy Caucasian subjects (3 men and 2 women). Each subject performed one rebreathing test at sea level (SL) and two tests at the Pyramid laboratory at Lobuche, Nepal, at the altitude of 5,050 m, 1 day after arrival (HA1) and after 24 days of sojourn (HA2). The effects of progressive isocapnic hypoxia were tested by using a standard rebreathing technique. BP, electrocardiogram, arterial oxygen saturation, airflow and end-tidal CO2 and O2 were recorded. For each subject, the relationships between arterial oxygen saturation and HR, systolic BP and minute ventilation (VE), respectively, were evaluated. At HA1, the majority of subjects showed a significant increase in VE and BP response and a decrease in HR response to progressive isocapnic hypoxia as compared to SL. At HA2, VE and BP responses further increased, whereas the HR response remained similar to that observed at HA1. A significant relationship between hypoxic ventilatory responses and both systolic and diastolic BP responses to progressive hypoxia was found. No significant correlation was found between hypoxic ventilatory and HR responses. PMID- 8727561 TI - Comparison of magnetic and electrical phrenic nerve stimulation in assessment of diaphragmatic contractility. AB - Unlike the standard electrical approach, cervical magnetic stimulation of the phrenic nerves is less painful and achieves a constant degree of diaphragmatic recruitment, features that should enhance its applicability in a clinical setting. An unexplained phenomenon is the greater transdiaphragmatic twitch pressure (Pditw) with magnetic vs. electrical stimulation. We hypothesized that this greater Pditw is due to coactivation of extradiaphragmatic muscles. Because impedance to rib cage expansion is increased at high lung volumes and efficiency of extradiaphragmatic muscles is less than that of the diaphragm, we reasoned that the difference between electrical Pditw and magnetic Pditw would be less evident at high volumes than at end-expiratory lung volume. In human volunteers, magnetic Pditw and electrical Pditw were 37.7 +/- 1.9 (SE) and 32.3 +/- 2.2 cmH2O, respectively, at end-expiratory lung volume (P < 0.005) and 24.0 +/- 2.9 and 27.2 +/- 2.8 cmH2O, respectively, at one-half inspiratory capacity (not significant); at total lung capacity, magnetic Pditw was less than electrical Pditw (10.6 +/- 0.8 and 16.2 +/- 2.9 cmH2O, respectively; P < 0.05). Magnetic stimulation caused significant extradiaphragmatic muscle depolarization and rib cage expansion, whereas electrical stimulation caused virtually no extradiaphragmatic muscle depolarization and rib cage deflation. Despite these differences, the induction of respiratory muscle fatigue produced reductions in both electrical and magnetic Pditw values (P < 0.01), which were of similar magnitude and closely correlated (r = 0.96). In conclusion, magnetic stimulation recruits both extradiaphragmatic and diaphragmatic muscles, and it is equally as effective as electrical stimulation in detecting diaphragmatic fatigue. PMID- 8727563 TI - Respiratory control during volume-cycled ventilation in normal humans. AB - The purpose of this study was to evaluate the extent of inhibition to respiratory rhythm associated with high volumes of ventilation during volume-cycled mechanical ventilation (neuromechanical inhibition). Two approaches were used. 1) In 18 normal awake subjects, ventilator tidal volume (VT) in the assist/control mode (A/C) was increased in steps from the minimum tolerable level up to 80% of the subject's inspiratory capacity or ventilator's maximum VT. We looked for appearance of intermittent apnea or a reduction in spontaneous rate (f). 2) Another 18 normal awake subjects were placed on controlled mechanical ventilation (CMV). When apnea was established, we abruptly terminated CMV and measured the time before the appearance of the next spontaneous effort. In the assist mode (protocol 1), we did not observe intermittent apnea because VT was increased from [from 944 +/- 198 to 1,867 +/- 277 (SD) ml], and there was only a modest reduction in f (14.1 +/- 3.9 to 12.4 +/- 4.0 breaths/min). End-tidal PCO2 (PETCO2) decreased precipitously as VT was increased. In protocol 2, we did not observe apnea after discontinuation of CMV in any subject. Total breath duration of the first breath after discontinuation did not differ significantly from total breath duration during A/C in the same subjects (4.84 +/- 2.2 vs. 5.2 +/- 2.0 s). This similarly applied regardless of route of breathing (nose vs. mouth) or PETCO2 level at time of discontinuation. We conclude that neuromechanical inhibition is quite weak and provides very little negative feedback that may help control PCO2 in the face of excessive VT and f demands of the subject. PMID- 8727562 TI - Determinants of basal fat oxidation in healthy Caucasians. AB - In a retrospective study, we examined several determinants of basal fat oxidation in 720 healthy Caucasian volunteers. Adult men (n = 427) and women (n = 293) were characterized for resting energy expenditure and substrate oxidation by indirect calorimetry (after a 12-h overnight fast), peak O2 consumption by a treadmill test to exhaustion, body composition by hydrodensitometry, food intake from a 3 day food diary, and hormonal status by fasting hormone concentrations. Fat oxidation was negatively correlated with fat mass in men (r = -0.11; P < 0.05), but no statistical relationship was found in women. In a stepwise multiple regression analysis, fat oxidation was best predicted by peak O2 consumption and fat-free mass in men (model R2 = 0.142) and by free thyroxine, fat-free mass, and fasting insulin in women (model R2 = 0.153). Relative rates of fat oxidation (fat oxidation adjusted for differences in resting energy expenditure) were not correlated with fat mass in either gender. Women showed a lower rate of basal fat oxidation (both absolute and adjusted) than did men. Our results show that fat oxidation is not greater in individuals with a greater fat mass. Furthermore, our results support a sexual dimorphism in basal rates of fat oxidation. PMID- 8727564 TI - Effect of N-methyl-D-aspartate-receptor blockade on hypoxic ventilatory response in unanesthetized piglets. AB - The central excitatory amino acid (EAA) neurotransmitter glutamate has been shown to mediate the ventilatory response to hypoxia through N-methyl-D-aspartate (NMDA) receptors in anesthetized adult animals. To determine the role of the EAA glutamate in the neonatal ventilatory response to hypoxia, 19 unanesthetized chronically instrumented piglets were studied. Minute ventilation (VE), oxygen consumption (VO2), arterial blood pressure (ABP), heart rate (HR), and blood gases were measured in room air (RA) and after 1, 5, and 10 min of hypoxia (inspired oxygen fraction = 0.10) before and after an infusion of saline or CGS 19755, a competitive NMDA-receptor blocker (10 mg/kg i.v.). Nine control piglets [age 6 +/- 1 (SD) days; weight 2.02 +/- 0.40 kg] and 10 CGS-19755-treated animals (age 6 +/- 1 days; weight 1.90 +/- 0.66 kg) were studied during quiet sleep and in a thermoneutral environment. There was a marked decrease in the VE response to hypoxia after the administration of CGS-19755. The ventilatory response to hypoxia was not modified by saline infusion. Changes in ABP and arterial PO2 during hypoxia were similar between groups, whereas the decrease in arterial PCO2 was significantly less after CGS-19755 administration. The increase in HR with hypoxia was eliminated by the NMDA-receptor blocker administration. VO2 decreased with hypoxia in both groups, but this decrease was more marked after the NMDA receptor blockade. These results suggest that the central EAA glutamate mediates, at least in part, the hypoxic hyperventilation in unanesthetized newborn piglets. PMID- 8727565 TI - Effects of exogenous surfactant on lung pressure-volume characteristics during liquid ventilation. AB - Total liquid ventilation (LV) lowers airway pressures and potentially reduces barotrauma in models of hyaline membrane disease. LV eliminates surface tension by eliminating the air-perfluorochemicals (PFC) interface but does not eliminate interfacial tension (IT) at the lung/PFC interface. We hypothesized that pretreatment with exogenous surfactant before LV would shift the overall pressure volume (PV) curve to the left, compared with LV without surfactant. Sequential quasi-static PV curves were obtained in 10 excised lungs (saline, air, PFC), with one-half randomized to exogenous surfactant replacement before LV. Analysis revealed that maximal inflation pressures were reduced during LV compared with baseline air curves. Addition of exogenous surfactant to LV further reduced maximal inflation pressures. A novel approach was used to transform these PV curves to estimates of in situ IT-volume curves. Estimated maximal IT at 20 ml/kg in preterm lamb lungs on air inflation after surfactant was 51 mN/m, compared with 40 mN/m for LV alone and with 27 mN/m for the combination of surfactant and LV. We conclude that the IT-reducing properties of the PFC studied (perflubron) can be augmented through the use of exogenous surfactant. PMID- 8727566 TI - Effects of pursed-lips breathing and expiratory resistive loading in healthy subjects. AB - To examine the effect of pursed-lips breathing (PLB) on breathing pattern and respiratory mechanics, we studied 11 healthy subjects breathing with and without PLB at rest and during steady-state bicycle exercise. Six of these subjects took part in a second study, which compared the effects of PLB to expiratory resistive loading (ERL). PLB was found to prolong expiratory and total breath durations and to promote a slower and deeper breathing pattern. During exercise, the compensatory increase that occurred in tidal volume was not sufficient to counter the reduction in breathing frequency, causing minute ventilation to be reduced. Although ERL similarly caused minute ventilation and breathing frequency to be decreased, unlike PLB, it produced no change in tidal volume and prolonged expiratory and total breath durations to a lesser extent. PLB and ERL increased the expiratory resistance to a comparable degree, also increasing the expiratory resistive work of breathing and promoting greater expiratory rib cage and abdominal muscle recruitment in response to the expiratory loads. End-expiratory lung volume, which was determined from inspiratory capacity maneuvers, was not altered by PLB; however, with ERL it was increased by 0.20 and 0.24 liter during rest and exercise, respectively. Inspiratory muscle recruitment patterns were not altered by PLB at rest, although small increases in the relative contribution of the rib cage/accessory muscles in conjunction with abdominal muscle relaxation occurred during exercise. Similar trends were observed with ERL. We conclude that, although ERL and PLB induce comparable respiratory muscle recruitment responses, they are not equivalent with respect to breathing pattern changes and effect on end-expiratory lung volume. PMID- 8727567 TI - Scalp topography of the short-latency components of the respiratory-related evoked potential in children. AB - Respiratory-related evoked potentials (RREPs) have been elicited by inspiratory occlusion and recorded at electroencephalographic (EEG) sites overlying the somatosensory cortex in adults. The present study was the first to be conducted in normal children and was designed to identify the scalp distribution of the early RREP components. EEG responses to occlusion were recorded from CZ-C3, CZ C4, and 17 sites referenced to the linked earlobes. The RREP was observed in all subjects in the CZ-C3 and CZ-C4 electrode pairs. The earlobe-referenced recordings revealed two RREP patterns. The P1 and N1 peaks were found in C3, C4, P3, P4, T3, and T4. The RREPs recorded from the F3, F4, F7, and F8 electrodes did not exhibit either the P1 or N1 peaks. A negative peak (NF) occurred approximately 13 ms after the P1 peak. The results show that the RREPs to inspiratory occlusions were present bilaterally but diminished greatly over midline sites. Furthermore, consistent with mechanically and electrically elicited somatosensory evoked potentials, the RREP displayed a polarity inversion over the central sulcus in the early component latency range. PMID- 8727568 TI - Pulmonary capillary pressure during exercise in horses. AB - The object of this study was to relate pulmonary capillary pressure to arterial and wedge pressures during exercise. Pulmonary vascular pressures were measured in six standardbred horses exercising at speeds equivalent to 75, 90, and 100% of maximal heart rate. Vascular pressures were measured with transducer-tip catheters and expressed relative to esophageal pressure. Pulmonary capillary pressure was estimated by the arterial-occlusion technique modified for exercise. Mean pulmonary arterial, capillary and wedge pressures increased from 30.5 +/- 6.3, 17.8 +/- 4.3, and 13.4 +/- 1.6 mmHg, respectively, at rest, to 70.5 +/- 5.2, 42.1 +/- 5.3, and 38.4 +/- 5.6 mmHg, respectively, at maximal exercise. The largest part of the increase occurred during the first level of exertion. With exercise, the pressure across the lung barely doubled at a time when the cardiac output would have increased at least fivefold. Thus the absolute resistance in both pre- and postcapillary segments must have decreased. The capillary and wedge pressures rose similarly, whereas the difference between them did not change with exertion. The fractional resistance of the precapillary segment increased with exercise. The postcapillary resistance, initially 28% of the total pulmonary vascular resistance, fell to 9% at maximal exercise. The rise (to approximately 45 mmHg) in pulmonary capillary pressure with exertion is consistent with an increase in transvascular filtration. PMID- 8727569 TI - Voluntary chronic exercise augments in vivo natural immunity in rats. AB - The effect of chronic voluntary exercise on the immune response was studied in spontaneously hypertensive rats. Exercise consisted of voluntary running in wheels for 5 wk, and the mean running distance was 4.2 km/24 h. In vivo cytotoxicity was measured as clearance of injected 51Cr-labeled YAC-1 lymphoma cells from the lungs. The clearance of YAC-1 cells in vivo was significantly increased in runners compared with sedentary controls (P < 0.001). The total number of mononuclear cells in the spleen was significantly decreased in runners compared with controls. Analysis of splenic lymphocyte phenotypes revealed a significantly increased fraction of OX52+/CD5- natural killer cells in runners compared with sedentary controls. In contrast to changes in natural immunity, immunoglobulins G and M levels in serum, the antibody response to antigen in vivo, and the proliferation of splenic T cells in vitro were unchanged. Our data suggest that chronic voluntary exercise augments natural cytotoxicity mechanisms in vivo, whereas splenic T-cell proliferation and the antibody-mediated immune response remain unchanged. PMID- 8727570 TI - Dilution acidosis: evidence for a role of intracellular pH in the control of ventilation. AB - Acute hyperosmolality results in an extracellular dilution acidosis and hypercarbia that does not stimulate ventilatory compensation. The osmotic stress is also associated with shifts in water and electrolyte balance and an increase in intracellular pH. The alkaline intracellular pH was hypothesized to have a role in preventing a normal respiratory response to the extracellular acidosis and hypercarbia. Therefore, this study examined the effect of ion-exchange blockade on intra- and extracellular pH and ventilation during acute hyperosmolality in the Pekin duck (Anas platyrhynchos) by using 31P-nuclear magnetic resonance spectroscopy. Both 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS) and amiloride inhibited the development of the intracellular alkalosis that normally develops in muscle during acute hyperosmolality. Instead, exposure to hyperosmotic stress during ion-exchange blockade resulted in a significant acidosis both intracellularly and extracellularly. Arterial pH decreased 0.10 +/- 0.04 pH unit with a sucrose infusion after either blocker, and intracellular pH decreased 0.11 +/- 0.06 and 0.16 +/- 0.04 pH units with a sucrose infusion after DIDS and amiloride, respectively. Ventilation increased 79 +/- 28 and 122 +/- 100%, respectively, during acute hyperosmolality after ion exchange blockade with either DIDS or amiloride. The results suggest that intracellular pH may play a role in the ventilatory response to acid-base perturbations. The data also indicate that both Cl-/HCO3- and Na+/H+ exchanges are involved in the development of the intracellular alkalosis during hyperosmotically induced extracellular acidosis. PMID- 8727571 TI - Ozone toxicity in the rat. II. Modeling changes due to ambient temperatures and duration. AB - Previous studies involving exposures to xenobiotic agents have demonstrated decreases in physiological parameters such as heart rate (HR) and core body temperature (Tco) and have shown that these toxic responses are modulated by changes in ambient temperature (Ta). We recently published the results of a study in which male Fischer 344 rats were implanted with radiotelemetry transmitters that permitted continuous monitoring of HR, Tco, and motor activity. These animals were divided into nine treatment groups (n = 4-5/group) composed of combinations of one of three O3-exposure regimens [0.0 parts/million (ppm) O3 x 24 h/day x 5 days; 0.5 ppm O3 x 6 h/day x 5 days; or 0.5 ppm O3 x 23 h/day x 5 days] at one of three Ta values (10,22, or 34 degrees C). We now report on statistical approaches for the modeling and analyses of these data. The models utilized were dependent on the treatment combinations. Circadian rhythms of Tco during air control periods were fit by cosine models. Overall effects of O3 in the 6- and 23-h exposure groups were best modeled by modified damped-sine and one compartment models, respectively, for both HR and Tco. These results demonstrate improved methods for the evaluation of biorhythmicity. PMID- 8727572 TI - On the existence of a lactate threshold during incremental exercise: a systems analysis. AB - The relationship between blood lactate concentration ([La]) and O2 uptake (VO2) during incremental exercise remains controversial: does [La] increase smoothly as a function of VO2 (continuous model), or does it begin to increase abruptly above a particular metabolic rate (threshold model)? The dynamic characteristics of the underlying physiological system are investigated using system identification analysis techniques. A multivariate deterministic time series model of the [La] and VO2 response to incremental changes in work rate was fitted to simulated and experimental data. Time-varying system response parameters were determined through the application of a weighted recursive least squares algorithm. The model, using the identified time-varying parameters, provided a good fit to the data. The variation of these parameters over time was then examined. Two major transitions in the parameters were found to occur at intensity levels equivalent to 53 +/- 8% and 77 +/- 9% maximal VO2 (experimental data). These changes in the model parameters indicate that the best linear dynamic model that fits the observed system behavior has changed. This implies that the system has changed its operation in some way, by altering its structure or by moving to a different operating region. The identified parameter changes over time suggest that the exercise intensity range (from rest to maximal VO2) is divided into three main intensity domains, each with distinct dynamics. Further study of this three-phase system may help in the understanding of the underlying physiological mechanisms that affect the dynamics of [La] and VO2 during exercise. PMID- 8727573 TI - Lumped-parameter tissue temperature-blood perfusion model of a cold-stressed fingertip. AB - A lumped-parameter model of a fingertip is presented. The semispherical model includes the effects of heat storage, heat exchange with the environment, and heat transport by blood perfusion. The thermal insulation on the surface of the fingertip is represented by the overall heat transfer coefficient that is calculated by common engineering formulas. The model is solved analytically for the simple case of constant blood perfusion rate. The general case of variable blood perfusion rates is solved by an Euler finite difference technique. At this stage, the model does not include active control mechanisms of blood perfusion. Thus the effects of cold-induced vasodilatation have to be superimposed and are modeled by symmetrical triangular waveforms because these were found to best depict the behavior of fingers exposed to cold environments. Results of this model were compared with experimental data obtained in two separate studies. One included 60-min infrared thermograms of the dorsal surface of bare hands of sedentary subjects horizontally suspended on a fish net in a 0 degree C environment. Another study, on gloved finger temperatures, involved 0 and -6.7 degrees C environments. Fingertip (nail bed) temperatures of both these studies were compared with model predictions. Blood perfusion rates were assumed and adjusted within physiologically reasonable limits. Comparison of measured and computed temperature records showed very good conformity in both cases studied. PMID- 8727574 TI - Use of a hypoxic lung as a deoxygenator to provide extended assessment of pulmonary function in rats. AB - Isolated perfused lung systems are commonly used to assess lung function in experimental studies. Assessment of hemodynamics and gas-exchange function in these systems is limited by the availability of venous blood. This study describes and validates a rat lung perfusion circuit in which a double-lung block ventilated with a hypoxic gas mixture [inspired O2 fraction (FIO2) 0.04; inspired CO2 fraction 0.08; deoxygenator (Deoxy) block] is used to provide blood with blood gases that are similar to mixed venous values to perfuse a study lung (FIO2 0.21; left lung only). This allows extended assessment of hemodynamics and gas exchange. Fifty adult male Wistar rats (300-400 g) were used as double-lung donors. Twenty-five perfusions (of both Deoxy and study lungs) were performed in four protocols (groups 1-5; n = 5). In protocol 1 (group 1), we tested whether exposure to room air affects the gas composition of the blood in the system. We found that the gas composition of the venous reservoir blood was identical to that of the blood entering the study block. In protocol 2, the effect of perfusion time and perfusion flow rate on the stability of the system was assessed. Lungs were perfused at 4 and 12 ml/min (groups 2 and 3, respectively), and the procedure was discontinued if edema or a marked decline in hemodynamics or gas-exchange function was observed. Pulmonary function was excellent and remained stable for 3 (at 12 ml/min) and 5 h (at 4 ml/min). In protocol 3, we examined whether hypoxic ventilation in the Deoxy lungs affects the stability of the system. Despite the low FIO2 used in the Deoxy lungs, the mean pulmonary arterial pressure-to-blood flow relationships in the study and Deoxy lungs were similar. Finally, in protocol 4, perfusion of a damaged study lung did not impair the function of the system. We conclude that this model permits reliable assessment of pulmonary function in rats under controlled ventilation and perfusion conditions. The use of a Deoxy double-lung block simplifies the perfusion apparatus and eliminates the main cause of instability of other systems that use an anesthetized host animal to provide venous blood. PMID- 8727575 TI - Airway inhomogeneities contribute to apparent lung tissue mechanics during constriction. AB - Recent studies have suggested that part of the measured increase in lung tissue resistance after bronchoconstriction is an artifact due to increased airway inhomogeneities. To resolve this issue, we measured lung impedance (ZL) in seven open-chest rats with the lungs equilibrated on room air and then on a mixture of neon and oxygen (NeOx). The rats were placed in a body box with the tracheal tube leading through the box wall. A broadband flow signal was delivered to the box. The signal contained seven oscillation frequencies in the 0.234- to 12.07-Hz range, which were combined to produce tidal ventilation. The ZL was measured before and after bronchoconstriction caused by infusion of methacholine (MCh). Partitioning of airway and tissue properties was achieved by fitting ZL with a model including airway resistance (Raw), airway inertance, tissue damping (G), and tissue elastance (H). We hypothesized that if the inhomogeneities were not significant, the apparent tissue properties would be independent of the resident gas, whereas Raw would scale as the ratio of viscosities. Indeed, during control conditions, the NeOx-to-air ratios for G and H were both 1.03 +/- 0.04. Also, there was a small increase in lung elastance (EL) between 0.234 and 4 Hz that was similar on air and NeOx. During MCh infusion, Raw and G increased markedly (45 65%), but the increase in H was relatively small ( < 13%). The NeOx-to-air Raw and H ratios remained the same. However, the NeOx-to-air G ratio increased to 1.19 +/- 0.07 (P < 0.01) and the increase in EL with frequency was now marked and dependent on the resident gas. These results provide direct evidence that for a healthy rat lung airway inhomogeneities do not significantly influence the lung resistance or EL vs. frequency data. However, during MCh-induced constriction, a large portion of the increase in tissue resistance and the altered frequency dependence of EL are virtual and a consequence of the augmented airway inhomogeneities. PMID- 8727576 TI - Frontonasal dysplasia: analysis of 21 cases and literature review. AB - Twenty-one patients with frontonasal dysplasia were studied. A 2:1 male-to-female sex ratio and increased paternal and maternal ages at the time of conception were found. The significance is uncertain because of small sample size and lack of normal mean values for parental age in Brazil. Apparently, our series is the first to report macrocephaly (six cases). Our series also had a high frequency of patients with agenesis of the corpus callosum (12 cases), basal encephalocele (10 cases), lipoma of the corpus callosum (four cases), and mental deficiency (11 cases). Three patients had the combination of agenesis of the corpus callosum, mental deficiency, and micropenis. It is concluded that frontonasal dysplasia is pathogenetically heterogeneous, representing a regional defect which may not be a single developmental field or sequence. Causal genesis includes a dominantly inherited form, dup(2q), and autosomal recessive Shanske syndrome. Of unknown genesis are two subsets of frontonasal dysplasia patients: 1) the combination of agenesis of the corpus callosum, tibial hypoplasia, and hallucal duplication and 2) ophthalmofrontonasal dysplasia or oculoauriculofrontonasal dysplasia with associated ear tags and epibulbar dermoids. PMID- 8727577 TI - Reconstruction of mandible and surrounding soft tissues in patient with necrotizing fasciitis. AB - Necrotizing fasciitis is rare in the orofacial region, with fewer than 20 cases reported in the literature. Extension of the disease process to involve the underlying bone has not been previously reported. A patient is presented in whom destruction of superficial skin and fascia, necrosis of a portion of the mandible, and involvement of the parotid gland complicated reconstruction. PMID- 8727578 TI - Generalized joint hypermobility as a factor in clicking of the temporomandibular joint. AB - Twenty-six patients with clicking or locking of the temporomandibular joint (TMJ) and 28 age- and sex-matched controls were assessed for generalized joint hypermobility by BEIGHTON's method. Function of the masticatory apparatus was measured with FRICTON's Craniomandibular Index. The TMJ patients had significantly higher joint mobility scores, but within the patient group there was no correlation between either the dysfunction or palpation scores and the hypermobility scores. The findings are interpreted as supporting multiple causes of TMJ internal derangement, one of which is a systemic joint factor. PMID- 8727579 TI - Palatal surgical splint for transverse stability of Le Fort I osteotomies: a technical note. AB - A modified surgical splint for Le Fort I osteotomies with transverse expansion is presented. The splint is made of a transpalatal stainless steel bar with acrylic abutment against the palatal surface of the molar and bicuspid tooth. It is rigid and renders excellent retention. It causes minimal patient discomfort, and oral hygiene is hardly compromised. PMID- 8727580 TI - Open reduction of subcondylar fractures. A study of functional rehabilitation. AB - Open reduction of dislocated subcondylar fractures was carried out in 19 adult patients between 1990 and 1992. A comparison of the results between two groups of patients was made 1 year after trauma. The reference group consisted of patients with the same type of fracture but treated with only intermaxillary fixation (IMF) and moderate jaw exercises after release of IMF. Dysfunction of the masticatory system was noticed in only a few cases of either group. Open reduction with plate osteosynthesis made it possible to avoid IMF in 12 of 19 patients. The results in the two groups did not differ significantly from a functional point of view. There are, however, specific indications for open reduction based on the degree of dislocation and concomitant subjective symptoms. PMID- 8727581 TI - One-step reconstruction of the severely resorbed mandible with onlay bone grafts and endosteal implants. A 5-year follow-up. AB - The severely resorbed mandibles (Cawood and Howell class VI) of 31 female patients were reconstructed with a corticocancellous bone graft with ITI implants in a one-step procedure. After 12-16 weeks, a combined mucosally and implant supported overdenture was made by an experienced prosthodontist. After 3, 6, 12, 24, and 60 months, orthopantomograms were made, and the height of the bone was measured mesial and distal of each implant. The overall bone resorption rate was almost 50%, but it varied around each implant. Eight of 78 implants were lost. Bone resorption occurred in an unpredictable way. The technique used can no longer be recommended. PMID- 8727582 TI - Conversion disorder after maxillofacial trauma and surgery. AB - Three cases of conversion disorder are reported: one after orthognathic surgery and two after facial trauma. Two of the cases involved facial anesthesia and one case involved facial palsy. It was thought to be important to bring this phenomenon to the attention of oral and maxillofacial surgeons, to consider its management, and to define the patients with a high risk of developing conversion disorder. The prognosis is dependent on the circumstances of the onset of the disorder, the premorbid psychologic health of the patient, and how rapidly treatment is prescribed. PMID- 8727583 TI - Oral presentation of secondary chondrosarcoma arising in osteochondroma of the nasal septum. AB - A case is described in which a midline palatal swelling, on incisional biopsy, showed an ossifying, chondroid lesion with the histologic features of a low-grade chondrosarcoma. Complete excision revealed that the tumour had originated in a benign osteochondroma of the nasal septum. PMID- 8727584 TI - Alveolar rhabdomyosarcoma (solid variant) of the maxillary sinus in an adult. PMID- 8727585 TI - Odontogenic keratocysts--a retrospective clinical and histologic study. AB - Including 25 recurrences, the material comprised 87 odontogenic keratocysts (OKCs) from 57 patients, who had been treated in 1974-93. The study included an analysis of clinical and histologic data from the patients' records and from pathologic reports. The men:women ratio was 1.5:1. Most of the patients belonged to the age groups 10-19, 20-29, 40-49, and 60-69 years. Almost 75% of the OKCs were localized in the mandible. The most common site was the angle and ascending ramus area. Recurrences comprised 30.5% of the OKCs. The symphysial area of the mandible had the highest recurrence rate (50%). The diagnosis of Gorlin's syndrome was established in two patients. The recurrence rates for the OKCs that were removed in one or in several pieces were 27.5% and 39%, respectively. In 14/87 (16%) cases, microkeratocysts were histologically found. Parakeratosis was observed in 78/87 (89.5%), ortho- and parakeratosis in 6/87 (7%), and orthokeratosis in 3/87 (3.5%) cases. The recurrence rates were 31.5% and 41% for the OKCs that were removed during 1974, 1984, and 1985-90, respectively. In no case could malignant changes be observed at histologic examination. PMID- 8727586 TI - Application of the buccal fat pad to the surgical treatment of oral submucous fibrosis. AB - The pedicled buccal fat pad has been widely used for the repair of oral defects. A new application of this flap in the treatment of patients suffering from trismus caused by oral submucous fibrosis is reported. The patients underwent incision of the fibrotic bands and coverage of the buccal defect with a pedicled buccal fat pad flap. The surgical technique is described, and the results suggest that this is a logical, convenient, and reliable technique for the treatment of oral submucous fibrosis. PMID- 8727587 TI - Granular cell tumor of buccal mucosa--a case report. PMID- 8727588 TI - Candida albicans: a review of its history, taxonomy, epidemiology, virulence attributes, and methods of strain differentiation. AB - The dimorphic yeast Candida albicans has been recognized as an increasingly important human pathogen particularly in immunocompromised hosts because of advanced age, infection or immunosuppressive therapy. This review outlines the history, taxonomy and epidemiology of this medically important yeast as well as discussing some of characteristics which are purported to be related to its virulence. Methods utilized for strain differentiation in the study of the epidemiologic relationship of members of this species are discussed. PMID- 8727589 TI - Probable BCG osteomyelitis of the hard palate: a case report. AB - A 4-year-old child with probable multifocal BCG osteomyelitis is reported. The lesions in the skull, clavicula, humerus, ribs, fibula, calcaneus, metatarsus, and hard palate were mainly osteolytic and healed rapidly with antituberculotic therapy. This is the first time that involvement of the hard palate has been described. PMID- 8727590 TI - Image-guided surgery. AB - Interventional video tomography (IVT), a new imaging modality, achieves virtual visualization of anatomic structures in three dimensions for intraoperative stereotactic navigation. Partial immersion into a virtual data space, which is orthotopically coregistered to the surgical field, enhances, by means of a see through head-mounted display (HMD), the surgeon's visual perception and technique by providing visual access to nonvisual data of anatomy, physiology, and function. The presented cases document the potential of augmented reality environments in maxillofacial surgery. PMID- 8727591 TI - The effect of gradual distraction of the mandible on the sheep temporomandibular joint. AB - The effect of gradual distraction of the mandibular ramus on the condylar process of the temporomandibular joint (TMJ) was studied in 17 growing sheep. Gradual lengthening affected both operated and contralateral sides. An enhancement of the endochondral ossification was noticed with marked osteoblastic activity between the cartilaginous layer and the bone formed. The cartilage was thinner and the bone structure more dense than in nonoperated controls, and woven bone predominated. These changes seemed to be partly reversible in the long term. PMID- 8727592 TI - Optimized growth medium for primary culture of human oral keratinocytes. AB - The effect of different media additives in defining optimal growth conditions for primary cultures of human oral keratinocytes was studied. A cocultivation technique with irradiated Swiss-3T3-fibroblasts in 96-well plates enables the comparison of additives for primary keratinocyte cultures derived from one patient. 3H-labeled thymidine uptake showed no growth or growth inhibition with adenine, choleratoxin or transferrin compared to basal medium (Dulbecco's modified Eagle's medium (DMEM) and 10% fetal calf serum). Among single additives, 5 micrograms/ml hydrocortisone, 5 micrograms/ml insulin, 10 ng/ml EGF, 2 micrograms/ml bovine pituitary extract, and 10(-9) M triiodothyronine showed the greatest capacity to promote keratinocyte growth. With all possible combinations of additives, maximum stimulation was found with a combination of EGF (10 ng/ml), insulin (5 micrograms/ml), and hydrocortisone (5 micrograms/ml); none of the other combinations were more effective. Our data indicate that in short-term cultures (up to 5 days) various media additives described in the literature are not necessarily required in this system of primary culture of human oral keratinocytes. PMID- 8727593 TI - An investigation of the interaction between alcohol and fibroblasts in wound healing. AB - This study investigated the effect of clinical concentrations of alcohol on fibroblast function (proliferation and ECM synthesis) in vitro. Basal and TGF beta-induced collagen synthesis was assayed in confluent cultures in serum-free medium at 48 h with a commercial collagen assay system. At concentrations of alcohol > 5%, fibroblast proliferation was significantly inhibited. Although noninhibitory, subclinical concentrations of alcohol failed to inhibit basal collagen synthesis (P > 0.1), they significantly decreased TGF-beta-induced collagen synthesis (P < 0.03). These data support the notion that the local, as well as the systemic, effects of alcohol are important in mediating delayed healing in alcoholic patients. PMID- 8727594 TI - ELI's coming: expression library immunization and vaccine antigen discovery. PMID- 8727595 TI - Is the Nef protein of HIV-1 required for pathogenesis? PMID- 8727596 TI - Genetics of coxsackievirus B cardiovirulence and inflammatory heart muscle disease. AB - Coxsackieviruses B (CVBs) are etiological agents of human inflammatory myocardial disease. The genetics of the coxsackieviral virulence phenotype in mice are now beginning to be understood with the availability of infectious cDNA copies of CVB genomes. Investigations to date with CVB3 and CVB4 have shown that sites within a non-translated region and in the capsid proteins can affect the virulence phenotype. The relative importance of these sites to expression of the phenotype remains unclear. PMID- 8727597 TI - Iron acquisition in the pathogenic Neisseria. AB - Pathogenic Neisseria species need to obtain iron from the host to grow in vivo. Several iron-transport systems are known, and regulation of Neisseria iron transport genes occurs via the transcriptional regulator Fur. There is evidence that the ability to transport iron is crucial to the survival of these organisms in vivo. PMID- 8727598 TI - Bacterial transferrin and lactoferrin receptors. AB - Pathogenic members of the Neisseriaceae and Pasteurellaceae express outer membrane receptor proteins involved in the direct assimilation of iron from the host glycoproteins transferrin and lactoferrin. The critical requirement of iron for growth suggests that this function is an important component of colonization and infection. A model describing this novel process is presented. PMID- 8727599 TI - Role of signalling and cytoskeletal rearrangements in the pathogenesis of Entamoeba histolytica. AB - Virulence of Entamoeba histolytica is characterized by intestinal tissue invasion, engulfment of host cells and the formation of liver abscesses. The actin-rich cytoskeleton of the amoeba allows rapid changes in morphology in response to signals from external stimuli. Cellular and molecular studies have described some of the proteins that participate in signalling and in cytoskeletal changes. PMID- 8727600 TI - New insights into the mechanisms of fungal pathogenesis in insects. AB - Entomopathogenic fungi are attracting attention as potential biological control agents of insect pests. The mechanisms of pathogenesis have parallels with those of some plant-pathogenic fungi, particularly in the areas of formation of an infection structure, entry into the host and toxin-mediated host death. Understanding these processes will provide a rational basis for strain selection and improvement. PMID- 8727601 TI - Epstein-Barr virus and the B cell: that's all it takes. AB - Recent experiments demonstrate that a much broader range of B cells harbor Epstein-Barr virus (EBV) in vivo than was previously expected from in vitro studies. In this review it is argued that EBV persists in vivo by integrating its biology with that of the normal B cells within which it resides, and that the B cell provides all the environments necessary for EBV to maintain its life cycle. PMID- 8727602 TI - Hepatocellular carcinoma. PMID- 8727603 TI - Pathogenesis of hepatocellular carcinoma: a review from the viewpoint of molecular analysis. AB - Chronic liver inflammation and hepatic regeneration by infection with hepatitis B (HBV) or C virus (HBC) seem to be important risk factors for hepatocellular carcinoma (HCC). Regarding the hepatocarcinogenesis of HBV DNA integration, it has been variously hypothesized that mechanisms such as the alteration of host chromosomal DNA and transcriptional trans-acting activity of the X gene are activated. On the other hand, integration of HCV virus into chromosomal DNA has not been reported. It is suggested that HCV could replicate more efficiently in noncancerous than in cancerous tissues. Therefore, it might affect some oncogenes or cause an inactivation of tumor suppressor genes in the early stage of HCC. Genetic alterations such as a point mutation and loss of heterozygosity are considered to be late events occurring after tumorigenesis. Regeneration of liver cells through chronic hepatitis increases the incidence of genetic alterations in hepatic cells and/or HCCs in both HBV- and HCV-infected patients. PMID- 8727604 TI - Usefulness of simultaneous determination of alpha-fetoprotein and des-gamma carboxy prothrombin in hepatocellular carcinoma. AB - Serum gamma-fetoprotein (AFP) and plasma des-gamma-carboxy prothrombin (DCP), a protein induced by vitamin K absence or antagonist II (PIVKA-II) levels, were measured in 197 patients with primary hepatocellular carcinoma (HCC). DCP levels were determined by conventional enzyme immunoassay kit (E-1023) and a newly developed high-sensitivity kit using the avidin-biotin complex method. Cut-off levels of AFP and DCP by the E-1023 kit and of DCP by the high-sensitivity kit were put at 100 ng/ml, 0.1 arbitrary unit (AU)/ml, and 0.004 AU/ml, respectively. Positive rate of AFP and DCP by the E-1023 kit and the high-sensitivity kit for HCC was 48%, 44%, and 57%, respectively. The positive rate by combination assay with AFP and DCP by the high-sensitivity kit increased up to 73%. There was no correlation between serum levels of AFP and those of plasma DCP. A significant correlation between tumor size and DCP levels was observed, but not with AFP. The postoperative disease-free survival rates of patients in the group with elevated levels of AFP and DCP were lower than those with normal levels of AFP and DCP. There were various patterns of change in the AFP and DCP levels at the time of recurrence compared with preoperative patterns. The combination assay of AFP and DCP levels is useful for the diagnosis, prognosis, and postoperative monitoring for recurrence of HCC. PMID- 8727605 TI - Radiologic diagnosis of hepatocellular carcinoma. AB - Recent developments in medical imaging technology and their clinical application for the diagnosis and management of hepatocellular carcinoma (HCC) are reviewed. Real-time ultrasound is effective for the screening of HCC in high-risk patients with liver cirrhosis. Information on blood flow is available by color Doppler or power Doppler technique. Dynamic computed tomography (CT) using a fast scanner and the modified contrast administration technique is useful for detection, differential diagnosis, and staging of HCC. Application of magnetic resonance (MR) imaging is increasing with rapid improvements in speed of image acquisition and resolution. Dynamic MR imaging and MR angiography have also been available. Diagnostic and interventional hepatic angiography are now performed less invasively, more safely, and more quickly with modern technologies of digital subtraction angiography (DSA) and catheters. CT during hepatic arteriography (CT arteriography) and arterial portography (CTAP) are used for evaluation of small focal nodules in the cirrhotic liver. PMID- 8727606 TI - Intracaval endovascular ultrasonography for malignant hepatic tumor: new diagnostic technique for vascular invasion. AB - Successful operation for hepatic tumor extending to the inferior vena cava (IVC) depends heavily on accurate preoperative imaging. We investigate the diagnostic value of intracaval endovascular ultrasonography (US) in the diagnosis of invasion to the IVC. A retrospective study of 26 consecutive patients of hepatic tumor with possible invasion of the IVC was performed using conventional imaging techniques from the right femoral vein with an 8-French, 20-MHz intravascular US. Nineteen of 26 cases were operated on and 15 cases were resected, including three cases of combined resection of the IVC. The sonographic criterion for IVC invasion was obliteration of a single echogenic layer of the IVC wall or intracaval tumor mass. The results of intracaval endovascular US were compared with those of CT and cavography. Vascular invasion was obtained in seven of 26 cases. Vascular invasion was confirmed by pathologic examination of five resected specimens, including two autopsy and two operative findings. The sensitivity, specificity, and overall accuracy of intracaval endovascular US for the diagnosis of the IVC invasion were 100%, 94.7%, and 96.1%, respectively. The values were 85.7%, 63.2%, and 69.2% for CT and 71.4%, 68.4%, and 69.2% for cavography, respectively. The intracaval endovascular US clearly visualized the IVC and established the presence and extent of tumor invasion. Intracaval endovascular US is a useful technique that can precisely evaluate the IVC for possible hepatic tumor invasion, especially when presence or extent of vascular invasion is not definitely established by conventional imaging techniques. PMID- 8727607 TI - Mechanism and prevention of ischemia-reperfusion injury of the liver. AB - Interruption of liver blood flow is often necessary as an operative technique in liver surgery. However, this procedure causes liver damage and can be a factor in postoperative liver failure. Interruption of oxygen and substrate supply and accumulation of metabolites contribute to a great variety of cellular and subcellular dysfunctions. Impairment of liver microcirculation occurs after reperfusion. It has been presumed that there is an imbalance between the activities of vasoconstrictors and vasodilators which would determine the vascular condition during ischemia and reperfusion phases. Some mediators are known to act as cytotoxic factors, especially after reperfusion following ischemia. The phenomenon in which organ damage becomes worse, even after reperfusion, is called reperfusion injury. Mediators released from accumulated polymorphonuclear neutrophils and activated Kupffer cells such as oxygen radicals and inflammatory cytokines are associated with ischemia-reperfusion injury of the liver. Regulation of these mediators will be a therapeutic necessity for this kind of liver injury in the future. PMID- 8727608 TI - Advances in hepatic resection and results for hepatocellular carcinoma. AB - The mortality and morbidity of hepatic resection for hepatocellular carcinoma (HCC) have decreased in recent years because of the various advances in hepatic resection. Various improvements are evident in dissecting apparatus, liver hepatic inflow clamp, cold hepatic perfusion technique, intraoperative ultrasonography, accurate assessment of hepatic function, autologous blood transfusion, and so on. Five-year survival after hepatic resection for HCC was reported at 26-59% in Eastern as well as Western series. The prognostic factors were portal invasion, multiplicity, serum alpha-fetoprotein level, tumor size, associated cirrhosis, age, alcohol abuse, histologic classification, DNA ploidy, and surgical margin. Segmental or lobar hepatic resection brought about better survival, especially in stage I and II patients. Effective adjuvant therapy should improve the diagnosis. PMID- 8727609 TI - Non-invasive loco-regional therapy for hepatocellular carcinoma. AB - It is important to understand characteristic features of hepatocellular carcinoma (HCC) for its treatment, namely most HCC is associated with liver cirrhosis and is characterized by multicentric development. Percutaneous ethanol injection therapy (PEIT) has preferably been performed in Japan for small HCC, less than 30 mm in diameter, because no difference was found in the survival rate between patients treated by PEIT and surgical resection. One of the most important factors affecting the prognosis of PEIT was regular follow-up after treatment. For advanced nodular HCC, larger than 30 mm in diameter, transcatheter arterial chemo-embolization has usually been carried out. The prognosis of arterial chemotherapy for highly advanced HCC has been improved remarkably by using an implantable injection port. Additionally, the prevention of HCC recurrence after initial successful treatment has become an urgent problem. PMID- 8727610 TI - Surgical treatment for hepatocellular carcinoma and concomitant esophagogastric varices. AB - Preventing a rupture of esophagogastric varices (EGV) is very important in aggressively treating hepatocellular carcinoma (HCC) in cirrhotic patients. We therefore performed simultaneous partial hepatic resection and direct interruption procedure on nine patients with HCC and concomitant EGV. Patients were selected on the basis of their stages of HCC and hepatic functional reserve. Postoperative hospital courses of all patients were uneventful. Six patients had recurrence of HCC and received non-surgical anti-tumor treatments. Only one patient had upper gastrointestinal bleeding at 18 months after operation, and the other eight patients have had no episodes of upper gastrointestinal bleeding during the follow-up period. The 5-year survival rate of these patients was 48%. This operative procedure is quite effective and is one of the treatments of choice for patients with less advanced HCC and concomitant risk of EGV. PMID- 8727611 TI - Pathology of early hepatocellular carcinoma: progression from early to advanced. AB - Based on the pathomorphologic studies of resected hepatocellular carcinoma (HCC) and biopsy materials from minute HCCs of the early stage, it has been clarified that the majority of minute HCCs around 1 cm in diameter are well differentiated. When tumors reach approximately 2 cm in diameter, approximately one-third of them display various combinations of more than two cancerous tissues of different histologic grades. In those HCCs, the areas of less-differentiated cancerous tissues are almost always surrounded by well-differentiated ones and diminish in size along with the increase of tumor size. Then, they are completely replaced by moderately to poorly differentiated cancerous tissues. Dedifferentiation phenomenon characterized by such a replacement of well-differentiated cancerous tissues by less-differentiated ones seems to be closely related to tumor proliferation in HCC. Thus, better therapeutic effects may be expected if HCCs are treated while they are well differentiated and are less aggressive. PMID- 8727612 TI - Registries in Japan: current status of hepatocellular carcinoma in Japan. Liver Cancer Study Group of Japan. AB - The Liver Cancer Study Group of Japan registered 11,379 patients with hepatocellular carcinoma (HCC) diagnosed from January 1, 1990 to December 31, 1991 in 536 hospitals throughout the country. This nationwide survey revealed the current status of HCC in Japan regarding the epidemiology, clinical characteristics, histopathological features, diagnosis, surgical and conservative treatments, and the outcome. The survival rates of the HCC patients who received hepatic resection, transarterial embolization (TAE), and percutaneous ethanol injection (PEI) were also calculated based on follow-up from January 1, 1987 to December 31, 1991. Three-year and 5-year survival rates of the patients who underwent hepatic resection were 57.5% and 40.8%, respectively, and those of TAE were 19.5% and 8.0%, respectively. Three-year survival rate of the patients with PEI was 53.2%. Cox's multivariate analysis showed that significant prognostic variables after partial hepatectomy were serum alpha-fetoprotein level, tumor size, number of tumors, associated liver cirrhosis, age, surgical curability, and portal involvement. PMID- 8727613 TI - Liver transplantation and hepatocellular carcinoma. AB - Liver transplantation for advanced hepatocellular carcinoma (HCC) continues to be a subject of debate mostly due to a uniformly high failure rate secondary to recurrence. However, accumulating experiences have enabled detailed analysis of the prognostic factors affecting patient outcome. Clinicopathological factors affecting survival have been reported to include tumor size, number, lobar distribution, vascular invasion, lymphnode metastasis, and histologic type such as fibrolamellar variant of HCC. Poor prognosis has been associated with advanced stage of the tumor, particularly pTNM stage IV. Many transplant centers now use a protocol of adjuvant chemotherapy with reasonable success, although experiences are limited. Better understanding in tumor biology, particularly of the role of immunosuppression affecting tumor growth, will provide further success in the treatment of HCC by liver transplantation. PMID- 8727614 TI - Prevention of nosocomial bloodstream infections: a national and international priority. PMID- 8727615 TI - Pediatric primary gram-negative nosocomial bacteremia: a possible relationship with infusate contamination. AB - OBJECTIVE: To evaluate the potential contribution of "extrinsic" contamination of intravenous fluids in hospital bacteremia and infection. DESIGN: Prospective cross-sectional survey of infusate contamination, December 1992 to December 1993. SETTING: A pediatric department (1,500 admissions per year) in a general, urban teaching hospital, serving low-income patients. SAMPLES AND PATIENTS: Infusate samples (0.5 to 1.0 mL) from the injection port used by the staff were taken for cultures from all febrile or septic patients in hospital wards. At least four samples were taken each day; if no febrile or septic patients were available, other patients were sampled at convenience. RESULTS: A 6.8% positive culture rate (87 contaminates in 1,277 infusates) was obtained, without significant differences among the wards. Gram-negative organisms were recovered from 56 samples (62.9%), mainly of the tribe Klebsielleae (56.1%). Coagulase-negative staphylococci were isolated in 30 samples (33.7%). There was no significant difference between the febrile-septic group and the asymptomatic group in the rate of infusate contamination (P = .59). In eight patients, the same organisms were recovered from infusate and blood culture. The overall bacteremia rate was 2.5 per 100 discharges. CONCLUSIONS: Compared to previous reports, higher infusate contamination rates and different organisms (mainly gram-negative) were observed. In hospitals of underdeveloped countries, nosocomial infection control frequently is disregarded. Infusate contamination may be common and could lead to gram-negative bacteremia. In such settings, it seems advisable to perform surveillance studies to identify infusate contamination, because a single infusate contamination could be a signal for an epidemic. PMID- 8727616 TI - Gram-negative bacteremia in open-heart-surgery patients traced to probable tap water contamination of pressure-monitoring equipment. AB - OBJECTIVE: To determine the cause(s) of an outbreak of gram-negative bacteremia (GNB) in open-heart-surgery (OHS) patients at hospital A. DESIGN: Case-control and cohort studies and an environmental survey. RESULTS: Nine patients developed GNB with Enterobacter cloacae (6), Pseudomonas aeruginosa (5), Klebsiella pneumoniae (3), Serratia marcescens (2), or Klebsiella oxytoca (1) following OHS; five of nine patients had polymicrobial bacteremia. When the GNB patients were compared with randomly selected OHS patients, having had the first procedure of the day (8 of 9 versus 12 of 27, P = .02), longer cardiopulmonary bypass (median, 122 versus 83 minutes, P = .01) or cross-clamp times (median, 75 versus 42 minutes, P = .008), intraoperative dopamine infusion (9 of 9 versus 15 of 27, P = .01), or exposure to scrub nurse 6 (6 of 9 versus 4 of 27, P = .001) were identified as risk factors. When stratified by length of the procedure, only being the first procedure of the day and exposure to scrub nurse 6 remained significant. First procedures used pressure-monitoring equipment that was assembled before surgery and left open and uncovered overnight in the operating room, whereas other procedures used pressure-monitoring equipment assembled immediately before the procedure. At night, operating rooms were cleaned by maintenance personnel who used a disinfectant-water solution sprayed through a hose connected to an automatic diluting system. Observation of the use of this hose documented that this solution could have contacted and entered uncovered pressure-monitoring equipment left in the operating room. Water samples from the hose revealed no disinfectant, but grew P aeruginosa. The outbreak was terminated by setting up pressure-monitoring equipment immediately before the procedure and discontinuing use of the hose-disinfectant system. CONCLUSIONS: This outbreak most likely resulted from contamination of uncovered preassembled pressure monitoring equipment by water from a malfunctioning spray disinfectant device. Pressure-monitoring equipment should be assembled immediately before use and protected from possible environmental contamination. PMID- 8727617 TI - Control of endemic glycopeptide-resistant enterococci. AB - OBJECTIVE: To evaluate the epidemiology of, and control measures for, vancomycin resistant Enterococcus (VRE) in a renal unit. DESIGN: A 3-month, prospective, prevalence culture survey of patients on a 24-bed renal unit. SETTING: A 975-bed community teaching hospital. PATIENTS: Patients admitted to the renal unit over a 3-month period. Patients identified with VRE were each matched with four patients without VRE isolated over the study period. INTERVENTIONS/CONTROL MEASURES: Resistant-organism barrier precautions. To eradicate carriage of VRE, two patients with VRE stool colonization were treated with 5 days of oral doxycycline (100 mg twice per day) and rifampin (300 mg/day). RESULTS: Seven patients with VRE (8 isolates) were identified. Five isolates were Enterococcus faecium (vancomycin MIC = 16 to 256 micrograms/mL), two were Enterococcus faecalis (MICs = 16 and 124 micrograms/mL), and one was Enterococcus gallinarum (MIC = 8.0 micrograms/mL). Eradication of carriage with VRE was accomplished in two patients treated with doxycycline and rifampin. In the final 30 days of the culture survey and at 9 months, there were no further patients with VRE identified. CONCLUSIONS: Resistant-organism precautions and elimination of patient carriage may be useful measures for controlling the spread of low-prevalence endemic vancomycin resistant Enterococcus. PMID- 8727618 TI - Nosocomial infections in Spain: results of five nationwide serial prevalence surveys (EPINE Project, 1990 to 1994). Nosocomial Infections Prevalence Study in Spain. AB - OBJECTIVE: To determine trends in rates of nosocomial infections in Spanish hospitals. DESIGN: Prospective prevalence studies, performed yearly from 1990 through 1994. SETTING: A convenience sample of acute-care Spanish hospitals. PARTICIPANTS AND PATIENTS: The number of hospitals and patients included were as follows: 1990, 125 hospitals and 38,489 patients; 1991, 136 and 42,185; 1992, 163 and 44,343; 1993, 171 and 46,983; 1994, 186 and 49,689. A core sample of 74 hospitals, which participated in all five surveys and included a mean of 23,871 patients per year, was analyzed separately. RESULTS: The overall prevalence rate of patients with nosocomial infections in the five studies was as follows: 1990, 8.5%; 1991, 7.8%; 1992, 7.3%; 1993, 7.1%; and 1994, 7.2%. The prevalence rate of patients with nosocomial infection in the core sample of 74 hospitals was 8.9%, 8.0%, 7.4%, 7.6%, and 7.6%, respectively (test for trend, P = .0001). Patients admitted to intensive-care units had a 22.8% prevalence rate of nosocomial infection in 1994. The most common nosocomial infections by primary site were urinary tract infection and surgical site infections, followed by respiratory tract infections and bacteremia. More than 60% of all infections were supported by a microbiological diagnosis. CONCLUSIONS: The EPINE project provides a uniform tool for performing limited surveillance of nosocomial infections in most Spanish acute-care hospitals. Its use helps to spread an accepted set of definitions and methods for nosocomial infection control in the Spanish healthcare system. The surveys indicated that the prevalence of nosocomial infections has been reduced over the last 5 years in a core sample of Spanish hospitals. PMID- 8727619 TI - Basic microbiologic support for hospital epidemiology. AB - The laboratory plays a major role in the epidemiology program's efforts to minimize nosocomial infections in healthcare institutions. This article will describe some of the interactions between the laboratory and the epidemiology program, and will identify resources and procedures that the laboratory needs to achieve epidemiologic goals. PMID- 8727620 TI - Nosocomial colonization and infection in persons infected with human immunodeficiency virus. AB - Nosocomial infections appear to be increased in patients with acquired immunodeficiency syndrome (AIDS), compared to individuals with asymptomatic infection due to human immunodeficiency virus (HIV). Risk factors for bacterial colonization and infection include immunosuppression, prior treatment with some antibiotics, increased hospitalizations with longer lengths of stay, greater exposure to invasive devices such as indwelling intravenous or urinary catheters, and the degree of immunosuppression. Data suggest that other infectious agents such as Pneumocystis carinii, Mycobacterium tuberculosis, Mycobacterium avium complex, and Cryptosporidium may be acquired in healthcare facilities. Diagnosis and management of nosocomial infections in HIV-infected persons may be complicated by an atypical presentation, increased rates of relapse following treatment, presence of multiple infections, and early discharge from the inpatient setting. Accurate assessment of nosocomial infections and outbreaks in the hospital is complicated by limited data on the risk of transmission of both traditional and unusual pathogens in this population. Furthermore, some patients may acquire nosocomial pathogens during their initial hospitalization and present later with infections that normally would be classified as community acquired. Therefore, there probably is an underestimation of current nosocomial infection rates, and perhaps "hospital-associated" or "healthcare-facility-associated" might be more accurate terms for these infections. PMID- 8727621 TI - The analysis of disease clusters, Part I: State of the art. AB - Public health professionals often are asked to investigate apparent clusters of human health events, or "disease clusters." A cluster is an excess of cases in space (a geographic cluster), in time (a temporal cluster), or in both space and time. This is part I of an introductory-level review of the analysis of disease clusters for physicians and health professionals concerned with infection surveillance in hospitals. It reviews the status of the field with the hope of expanding the use of cluster analysis methods for the routine surveillance of infectious disease in the hospital environment. PMID- 8727622 TI - Mad cows and CJD. PMID- 8727623 TI - Terrorist access to biologic agents. PMID- 8727624 TI - Research can be a pain in the neck. PMID- 8727625 TI - To treat or not to treat (out-of-hospital) paroxysmal supraventricular tachycardia, that is the question. PMID- 8727626 TI - Another nail in the coffin... PMID- 8727627 TI - The effect of soft cervical collars on persistent neck pain in patients with whiplash injury. AB - OBJECTIVE: To assess the efficacy of soft cervical collars in the early management of whiplash-injury-related pain. METHODS: A controlled, clinical trial was conducted in an urban ED. Adults with neck pain following automobile crashes indicated their initial degrees of pain on a visual analog scale. Patients with cervical spine fractures or subluxation, focal neurologic deficits, or other major distracting injuries were excluded. Patients were assigned to receive a soft cervical collar or no collar based on their medical record numbers. Pain at > or = 6 weeks postinjury was coded as none, better, same, or worse, and analyzed as 3 dichotomous outcomes: recovered (pain = none); improved (pain = none or better); and deteriorated (pain = worse). RESULTS: Of 250 patients enrolled, 196 (78%) were available for follow-up. Of these patients, 104 (53%) were assigned to the soft cervical collar group, and 92 (47%) to the control group. These groups were similar in age, gender, seat position in the car, seat belt use, and initial pain score. Pain persisted at > or = 6 weeks in 122 (62%) patients. The groups showed no difference in follow-up pain category (p = 0.59). There was no significant difference between the 2 groups in complete recovery (p = 0.34), improvement (p = 0.34), or deterioration (p = 0.60). The study had a power of 80% to detect an absolute difference of at least 20% in recovery, 17% in improvement, and 7% in deterioration (2-tailed, alpha = 0.05). CONCLUSIONS: Most patients with whiplash injuries have persistent pain for at least 6 weeks. Soft cervical collars do not influence the duration or degree of persistent pain. PMID- 8727629 TI - Directional-tip endotracheal tubes for blind nasotracheal intubation. AB - OBJECTIVES: To compare initial success rates of blind nasotracheal intubation using directional-tip endotracheal tubes vs standard endotracheal tubes. METHODS: A prospective trial comparing directional-tip and standard endotracheal tubes during initial attempts at blind nasotracheal intubation (BNTI) at a university hospital ED. Using an alternating schedule, the directional-tip or standard tube was used for the first attempt at BNT1. An attempt was defined as beginning when the tube was placed through the nose into the posterior pharynx and ending when the patient was intubated or the tube was removed from the nares. After the intubation, the physician graded the difficulty of the technique (i.e., easy/routine, intermediate, difficult, or unable to intubate nasally). RESULTS: There were 49 patients entered over 5 months. Patient presentations for the intubations were trauma (45.8%), overdose (33.3%), respiratory/cardiac event (12.5%), seizure (2.1%), and other (6.3%). Intubation was successful on the first attempt in 18 of 21 patients (86%; 95% CI, 64% to 97%) for the directional-tip tube vs 16 of 28 patients (57%; 95% CI, 37% to 76%) for the standard tube (p = 0.03). The groups did not differ in age, sex, clinical presentation, or perceived difficulty of intubation. Only 1 patient could not be intubated nasally and was subsequently intubated orally. CONCLUSION: The use of directional-tip tubes may improve the success rate of the first attempt at BNTI. PMID- 8727628 TI - Treatment of out-of-hospital supraventricular tachycardia: adenosine vs verapamil. AB - OBJECTIVE: To compare the use of adenosine and the use of verapamil as out-of hospital therapy for supraventricular tachycardia (SVT). METHODS: A period of prospective adenosine use (March 1993 to February 1994) was compared with a historical control period of verapamil use (March 1990 to February 1991) for SVT. Data were obtained for SVT patients treated in a metropolitan, fire-department based paramedic system serving a population of approximately 1 million persons. Standard drug protocols were used and patient outcomes (i.e., conversion rates, complications, and recurrences) were monitored. RESULTS: During the adenosine treatment period, 105 patients had SVT; 87 (83%) received adenosine, of whom 60 (69%) converted to a sinus rhythm (SR). Vagal maneuvers (VM) resulted in restoration of SR in 8 patients (7.6%). Some patients received adenosine for non SVT rhythms: 7 sinus tachycardia, 18 atrial fibrilation, 7 wide-complex tachycardia (WCT), and 2 ventricular tachycardia; no non-SVT rhythm converted to SR and none of these patients experienced an adverse effect. Twenty-five patients were hemodynamically unstable (systolic blood pressure < 90 mm Hg), with 20 receiving drug and 13 converting to SR; 8 patients required electrical cardioversion. Four patients experienced adverse effects related to adenosine (chest pain dyspnea, prolonged bradycardia, and ventricular tachycardia). In the verapamil period, 106 patients had SVT: 52 (49%) received verapamil (p < 0.001, compared with the adenosine period), of whom 43 (88%) converted to SR (p = 0.11). Two patients received verapamil for WCT; neither converted to SR and both experienced cardiovascular collapse. VM resulted in restoration of SR in 12 patients (11.0%) (p = 0.52). Sixteen patients were hemodynamically unstable, with 5 receiving drug (p = 0.005) and 5 converting to SR; 9 patients required electrical cardioversion (p = 0.48). Four patients experienced adverse effects related to verapamil (hypotension ventricular tachycardia, ventricular fibrillation). Recurrence of SVT was noted in 2 adenosine patients and 2 verapamil patients in the out-of-hospital setting and in 23 adenosine patients and 15 verapamil patients after ED arrival, necessitating additional therapy (p = 0.48 and 0.88, for recurrence rates and types of additional therapies, respectively). Hospital diagnoses, outcomes, and ED dispositions were similar for the 2 groups. CONCLUSION: Adenosine and verapamil were equally successful in converting out-of-hospital SVT in patients with similar etiologies responsible for the SVT. Recurrence of SVT occurred at similar rates for the 2 medications. Rhythm misidentification remains a common issue in out-of-hospital cardiac care in this emergency medical services system. PMID- 8727630 TI - Serial myoglobin levels for patients with possible myocardial infarction. AB - OBJECTIVES: To determine the sensitivity and specificity of a new myoglobin assay for acute myocardial infarction (AMI), considering both the total amount of serum myoglobin and its percentage change over 2 hours. METHODS: A prospective, observational test performance study for the recognition of AMI was done using serial myoglobin assays of 42 admitted chest pain patients at a large, urban teaching hospital ED. Myoglobin testing was performed at presentation (time 0) and at 1 and 2 hours after arrival. A myoglobin level > 100 micrograms/L (ng/mL) or a change > or = 50% from baseline (increase or decrease) any time during the 2 hour period was considered positive. Patients and their physicians were blinded to the myoglobin results. The managing clinician's final diagnosis of the presenting event was used as the diagnostic criterion standard. RESULTS: The sensitivity of the myoglobin technique for detection of AMI in the first hours in the ED was 13/14 (93%; 95% CI: 66-100%). The 1 patient who had a false-negative test had evidence of AMI on the ECG and an initially abnormal creatine kinase-MB (CK-MB) assay. The specificity was 22/28 (79%; 59-92%). However, of the 6 patients who had "false-positive" myoglobin tests, all had serious illness: significant cardiac disease (n = 4), in-hospital death (n = 1), or deep venous thrombosis (n = 1). CONCLUSION: Myoglobin level determinations are sensitive tests to detect AMI during the first 2 hours of a patient's stay in the ED and may complement current clinical tools. PMID- 8727631 TI - Ketamine safety profile in the developing world: survey of practitioners. AB - OBJECTIVE: To evaluate the safety profile of ketamine when used to facilitate surgical procedures in the less controlled setting of developing world, rural hospitals. METHODS: A survey addressing clinical experience with ketamine in the developing world was administered to a convenience sample of missionary physicians. Descriptive statistics are reported. RESULTS: Of the 172 surveyed physicians, 122 (71%) responded; 55 reported experience with ketamine. These physicians estimated a total of 12,844 administrations. The format of one procedural physician and a second trained anesthesiologist/anesthetist was unavailable in the practice of 59% of the responding physicians, and 34% routinely performed procedures while simultaneously supervising ketamine administration and monitoring its clinical effect. Pulse oximetry was used "often" or "always" by only 10% of the physicians. Cardiac monitoring and intermittent vital signs were used in only 19% and 45%, respectively. One unexplained pediatric death occurred during an unmonitored, unobserved ward recovery. An adult suffered cardiac arrest after a failed intubation attempt. Seventeen other complications possibly related to ketamine were apnea (n = 10), laryngospasm (n = 6), and aspiration (n = 1), all of which were transient and without sequelae. Physicians believed that recovery hallucinations and agitation were frequent in adults and unusual in children. CONCLUSIONS: Death and other serious complications were rare in this survey reporting > 12,000 estimated ketamine administrations in the developing world. Although the limitations of survey data are recognized, the margin of safety with ketamine appears to be high, even when administered by non-anesthesiologists in settings lacking basic mechanical monitoring. These findings have important implications for the use of ketamine outside the controlled operating room environment in developed countries. PMID- 8727632 TI - Effect of time of day and duration into shift on hazardous exposures to biological fluids. AB - OBJECTIVE: To determine whether hospital employee biological hazardous exposure rates varied with time of day or increased with time interval into shift. METHODS: This was a retrospective occurrence report review conducted at a university hospital with an emergency medicine residency program. Health care worker biological hazardous exposure data over a 30-month period were reviewed. Professional status, date, time, and type of exposure (needlestick, laceration, splash), time interval into shift of exposure, and hospital location of exposure were recorded. Hourly employee counts and risky procedure counts were matched by location with each reported exposure, to determine hourly rates of biological hazardous exposures. RESULTS: Analysis of 411 recorded exposures demonstrated that more people were exposed between 9:00 AM and 11:00 AM (p < 0.05), yet the exposure risk did not vary significantly when expressed as the number of exposures per worker or per procedure. Of the 393 exposures with data describing time interval into shift when the exposure occurred, significant numbers of exposures occurred during the first hour and at shift's end [when corrected for exposures per worker (p < 0.05) or exposures per procedure (p < 0.05)]. CONCLUSION: While the number of exposures are increased in the AM hours, the exposure rate (as a function of workers or procedures) does not vary with time of the day. However, the exposure rate is increased during the first hour and last 2 hours of a shift. Efforts to increase worker precautions at the beginning and end of shifts are warranted. PMID- 8727633 TI - Posttraumatic multiple-organ dysfunction syndrome: role of mediators in systemic inflammation and subsequent organ failure. PMID- 8727634 TI - Emergency medicine expeditions to the developing world: the Loma Linda University experience in Papua New Guinea. AB - Emergency physicians (EPs) are increasingly participating in international medicine in regions that are chronically medically underserved. In August 1994, a ten-member emergency medicine team from the Loma Linda University School of Medicine staffed a 70-bed bush hospital in the primitive highlands of Papua New Guinea, providing both outpatient and inpatient medical care. Typhoid fever, malaria, polio, and numerous other infectious diseases were encountered. Rampant local tribal warfare resulted in regular penetrating injuries from arrows, spears, and machetes. The expedition was judged highly successful, in that 1) substantial medical service was provided to tribespeople accustomed to minimal care, 2) education was provided to local health care providers, and 3) team participants became adept at managing medical conditions uncommon in industrialized societies, and gained valuable ethical and utilitarian perspectives regarding health care delivery in underserved areas. In this article the objectives, organization, and experiences of the team members are described. This information may encourage other EPs to participate in medical expeditions to the developing world, and to provide general principles to assist in their organization and implementation. PMID- 8727635 TI - Pharmacist with hypotension and altered mental status. PMID- 8727636 TI - Ventricular fibrillation with background pacing masquerading as pulseless electrical activity. PMID- 8727637 TI - Hypertensive urgency concern. PMID- 8727638 TI - Grassroots emergency medicine: a survey of U.S. medical schools. PMID- 8727639 TI - The physician as hero. PMID- 8727641 TI - Phospholipid composition of neonatal guinea pig liver and plasma: effect of postnatal food restriction. AB - Preterm guinea pigs were delivered on day 65 of gestation (term = 68 d) and were allowed either free or restricted access to food for the subsequent 48 h. Plasma phosphatidylcholine (PC) concentration increased postnatally from 190 (range 144 307) to 751 (426-1039) and 883 (758-977) microM for fed and starved pups, respectively. Plasma PC composition in both groups of pups was characterized by selective and equivalent relative increases to individual molecular species containing 18:0 at the sn-1 position. Hepatic PC concentration increased from 6.75 (5.41-8.20) to 8.65 (6.54-10.63) and 9.23 (8.18-10.17) mumol/g for fed and starved pups, respectively, and, under all conditions, hepatic PC molecular composition closely mirrored that of plasma PC. These results support the hypothesis that the molecular species composition of plasma PC for the guinea pig in the immediate postnatal period is determined largely by the composition of the hepatic PC pool destined for lipoprotein secretion. Hepatic PC composition and concentration of the starved neonatal guinea pig were maintained independently of any dietary nutrient intake, at the expense of mobilization of extra hepatic lipid reserves. While this adaptive mechanism has inherent limited survival potential in neonatal starvation, it has implications for studies measuring plasma phospholipid fatty acid compositions as biochemical markers of dietary fat intake in preterm infants. PMID- 8727642 TI - Dietary triacylglycerol structure and saturated fat alter plasma and tissue fatty acids in piglets. AB - Human and pig milk triacylglycerols contain a large proportion of palmitic acid (16:0) which is predominately esterified in the 2-position. Other dietary fats contain variable amounts of 16:0, with unsaturated fatty acids predominantly esterified in the 2-position. These studies determined if the amount or position of 16:0 in dietary fat influences the composition or distribution of liver, adipose tissue, lung, or plasma fatty acids in developing piglets. Piglets were fed to 18 d with sow milk or formula with saturated fat from medium-chain triglyceride (MCT), coconut or palm oil, or synthesized triacylglycerols (synthesized to specifically direct 16:0 to the 2-position) with, in total fatty acids, 30.7, 4.3, 6.5, 27.0, and 29.6% 16:0, and in 2-position fatty acids, 55.3, 0.4, 1.3, 4.4, and 69.9% 16:0, respectively. The percentage of 16:0 in the 2 position of adipose fat from piglets fed sow milk, palm oil, and synthesized triacylglycerols were similar and higher than in piglets fed MCT or coconut oil. Thus, the amount, not the position, of dietary 16:0 determines piglet adipose tissue 16:0 content. The effects of the diets on the plasma and liver triacylglycerols were similar, with significantly lower 16:0 in total and 2 position fatty acids of the MCT and coconut oil groups, and significantly higher 16:0 in the plasma and liver triacylglycerol 2-position of piglets fed the synthesized triacylglycerols rather than sow milk or palm oil. The lung phospholipid total and 2-position 16:0 was significantly lower in the MCT, coconut, and palm oil groups, but similar in the synthesized triacylglycerol group and sow milk group. The lung phospholipid total and 2-position percentage of arachidonic acid (20:4n-6) was significantly lower in all of the formula-fed piglets than in milk-fed piglets. The physiological significance of this is not known. PMID- 8727644 TI - Changes in lipid composition during metamorphosis of bonefish (Albula sp.) leptocephali. AB - During metamorphosis of bonefish (Albula sp.) larvae (leptocephali) all energy requirements are provided by breakdown of endogenous compounds, with lipid catabolism accounting for about 80% of total energy production. The principal objective of the present study was to characterize the lipid classes and fatty acids utilized. Analysis of whole-body lipid content indicated that larvae lost about half (3.6 mg) of their total lipid during the 10-d period. Percentages of neutral and polar lipid in early metamorphosing larvae were 64.2 and 35.8%, respectively; these values showed little change during metamorphosis, indicating that both lipid classes were catabolized. Triacylglycerols, the principal neutral lipid of all metamorphic stages, decreased by 1.8 mg, accounting for half of the decrease in total lipid. Levels of phosphatidylethanolamine, the principal polar lipid in early larvae, decreased by more than 50% during metamorphosis; levels of phosphatidylcholine, which was not detected in early larvae, increased. Fatty acids showing the largest net decreases, presumedly used as energy sources, were 16:0 (30.4%), 14:0 (13.8%), 16:1n-7 (12.2%), 20:5n-3 (7.7%), 18:1n-9 (7.4%), and 18:4n-3 (6.9%). Most of 22:6n-3, the second most abundant fatty acid of early larvae, was conserved. PMID- 8727643 TI - Rat gastric hydrophobic barrier: modulation of phosphatidylcholine molecular species by dietary lipids. AB - Phospholipids protect the gastric mucosa by forming a proton-repellent hydrophobic layer on its luminal surface. We have recently shown that two molecular species of phosphatidylcholine (PC), PC16:0/18:1, and PC16:0/18:2, but not PC16:0/16:0, are predominantly released into gastric mucus. We investigated whether these molecular species in mucus are modified by dietary fat. Rats were fed (for three weeks) a diet supplemented with either 10% cod liver, palm, or sunflower oil, or 10% corn starch as a control. In tissue, cod liver oil decreased PC16:0/20:4 and PC18:0/20:4. Cod liver oil and palm oil increased PC16:0/18:1, whereas sunflower oil decreased PC16:0/18:1. Palm oil additionally decreased PC16:0/18:2, whereas the other diets had no effect on PC16:0/18:2. In mucus, however, PC16:0/18:1 and PC16:0/18:2 were not significantly altered by any diet. They were increased over tissue values and comprised 37.6 +/- 3.3 and 33.1 +/- 1.4 mol% in controls. PC16:0/16:0 was lower in mucus than in mucosa and even decreased by cod liver oil (1.2 +/- 0.2 vs. 2.7 +/- 0.3 mol%; P < 0.01). We conclude that PC16:0/18:1 and PC16:0/18:2 are modified by dietary fat in tissue. In gastric secretions, however, PC16:0/18:1 and PC16:0/18:2 are kept constant and together comprise 70 mol% of the released PC species, whereas PC16:0/16:0 does not play a role for the gastric hydrophobic barrier under any dietary treatment. Additionally, cod liver oil decreases the content of PC16:0/20:4 and PC18:0/20:4 in gastric mucosa, thereby possibly decreasing the formation of eicosanoids. PMID- 8727645 TI - Purification and characterization of deep sea shark Centrophorus squamosus liver oil 1-O-alkylglycerol ether lipids. AB - The 1-O-alkylglycerol composition of the liver oil of the deep sea shark Centrophorus squamosus, a species which provides edible flesh, has been determined. After various fractionations of the oil, the unsaponifiable fraction was characterized by means of gas chromatography/mass spectrometry, electron impact, and positive-ion chemical ionization. The oil is composed of 60% unsaponifiable matter, containing 45% squalene, 4.5% cholesterol, and 10% of linear saturated and monounsaturated glycerol ethers with 14-18 carbon atoms. After a first separation by chromatography on silicic acid, monounsaturated glycerol ethers have been separated from the saturated homologues, in particular from 1-O-octadecylglycerol (batyl alcohol) and 1-O-hexadecylglycerol (chimyl alcohol), via urea complexation. This newer application of the urea method, already used in the past to extract saturated from polyunsaturated fatty acids, allowed the purification of the main components of the complex unsaturated glycerol ether fraction, namely, 1-O-octadecen-9'ylglycerol (selachyl alcohol) and 1-O-hexadecen-9'ylglycerol. PMID- 8727646 TI - Microscale preparation of natural and labeled oxysterols. AB - The preparation of cholesterol and radiocholesterol oxidation products on a microscale is difficult. Cholesterol generally resists oxidation unless it is well dispersed under controlled conditions. A method was developed to reliably produce 7 alpha- and 7 beta-hydroxycholesterol and 7-ketocholesterol. Small changes in pH, metal ions present, or in the colloidal dispersion, resulted in production of completely different oxysterols. Attempts to oxidize aged radiocholesterol were not successful even after purification by several thin layer chromatographic steps, and this appeared to be due to a time-related change in the radioactive material. Fresh radiocholesterol oxidized readily. PMID- 8727648 TI - Synthesis of 1,2-diacyl-sn-glycerophosphatidylserine from egg phosphatidylcholine by phosphoramidite methodology. AB - A simple chemical method for the synthesis of 1,2-diacyl-sn glycerophosphatidylserine (PS), with the same fatty acid composition in the sn-1 and sn-2 glycerol positions as egg phosphatidylcholine (PC), is described. PS synthesis was carried out by a phosphite-triester approach, using 2-cyanoethyl N,N,N',N'-tetraisopropylphosphorodiamidite (phosphoramiditate) as the phosphorylating agent, for the formation of phosphate linkage between serine and diacylglycerol. 1,2-Diacylglycerol, obtained from PC hydrolysis by phospholipase C, was coupled with N-t-BOC-L-serinebenzhydryl ester phosphoramidite with tetrazole as catalyst. Phosphite-triester was oxidized to the corresponding phosphate-triester with 30% H2O2 in CH2Cl2. The cyanoethyl group was removed by addition of an Et3N/CH3 CN/pyridine mixture, and trifluoroacetic acid was used to eliminate the protecting groups of O-(1,2-diacylglycero-3-phospho)-N-t-BOC serinebenzhydryl ester. Purified PS was identified by thin-layer chromatography, infrared, and 1H nuclear magnetic resonance. PMID- 8727647 TI - An improved method for rapid analysis of the fatty acids of glycerolipids. AB - An improved rapid procedure to determine the fatty acid composition of glycerolipids is described. The procedure includes KOH-catalyzed transesterification and high-speed gas chromatography. Glycerolipids (20-40 mg) were mixed with 2 mL of hexane and 0.2 mL of 2 M methanolic KOH at room temperature for 1-2 min. The fatty acid methyl esters in the hexane layer were analyzed by gas chromatography on 10% SP-2340 at 240 degrees C. Methyl linolenate and docosahexaenoate eluted within 2 and 5 min, respectively. Analysis was thus completed within 5 min for common vegetable oils and 8 min for fish oils. PMID- 8727649 TI - Effect of medium-chain triglycerides on calbindin-D9k expression in the intestine. AB - These studies determined the effect of the saturated fat source in infant formula on the expression of calbindin-D9k (CaBP-9k). Piglets were fed from birth to 8 d with milk or formula containing saturated fatty acids as medium-chain triglycerides (MCT), coconut oil, palm oil (Palm 1), or synthesized triglycerides with 16:0 directed to the sn-2 position (Palm 2). Levels of intestinal CaBP-9k mRNA were significantly (P < 0.01) higher in piglets fed formula with MCT than in piglets fed the other formula or milk; and higher in piglets fed the Palm-1 than in piglets fed Palm-2 formula. This is the first evidence that MCT alter piglet intestinal CaBP-9k mRNA. PMID- 8727650 TI - A self-efficacy scale for HIV risk behaviors: development and evaluation. AB - To measure self-efficacy with respect to risk behaviors for HIV infection, a set of questionnaire items was developed regarding condom use, drug use with friends, and negotiations with potential sex partners. These items were tested with a sample of Latinos aged 14 to 22 years in two New England cities. A nine-item self efficacy scale was found to have a high level of internal consistency (Cronbach's alpha = .77). Reliability coefficients were similar for men, women, and both English- and Spanish-speaking respondents. Strong associations with recent performance accomplishments, as specified in Bandura's social cognitive theory, support the construct validity of the scale. PMID- 8727651 TI - HIV risk behaviors: a comparison of U.S. Hispanic and Dominican Republic youth. AB - Sexual behaviors of adolescents place them at a high risk for HIV infection and other sexually transmitted diseases. International comparisons of sexual risk taking among youth employing a common set of survey items are rare. Moreover, such studies can assist in explaining observed differences in disease rates, or predicting future trends in transmission, and identifying health educational needs unique to a particular population or setting. To assess variations in risk taking patterns, a comparison of HIV behavioral risk factors was made between U.S. Hispanic and Dominican Republic (D.R.) youth. The Spanish version of the Youth Risk Behavior Surveillance (YRBS) survey was administered to 1,175 D.R. high school students. Responses were compared to those in the U.S. from the Centers for Disease Control and Prevention's 1990 YRBS data tape for 2285 Hispanic youth. Frequency and cross-tabulation data were compared by country and gender. There were significant differences in AIDS education at school, discussions about AIDS conducted in the home, age of first sexual experience, condom use, and past experience with STDs. Results show that adolescents in both countries are at risk, but with different mediating factors. Inadequate condom use and early initiation of sex place the D.R. youth at risk, while more U.S. youth are engaging in sexual intercourse and using alcohol or other drugs in conjunction with sexual episodes. Some implications for prevention education are discussed. PMID- 8727652 TI - Japanese risk behaviors and their HIV/AIDS-preventive behaviors. AB - This study is to identify the risk behaviors of the Japanese that may lead to HIV infection and the behaviors that prevent such infection, as well as their background factors. Two behavioral surveys were conducted for the present study. (1) For international comparison on knowledge, attitudes, beliefs, and practices (KABP) related to HIV/AIDS, we conducted a survey on a sample of 10,000 adults, randomly selected from a nationwide population in Japan; and (2) for sexual partner relation, we conducted a survey on a sample of 10,000 adults randomly selected from a population in five major cities of Japan. Our main findings include: (1) Most of the Japanese adult did not regard AIDS as a major threat in the area where they lived; (2) People in their twenties are too casual about "having sex without using a condom with someone they've met for the first time and know little about"; (3) Thirteen percent (19% male, 8% female) of those with a steady sex partner including a spouse, on average, had sex with 2.4 non-steady partners in the previous year; and (4) Only 25 percent used condoms always when they engaged in casual sex during the previous four weeks. These risk behaviors of the Japanese adults might lead to an explosive rise in the number of HIV infected in the near future unless steps are taken immediately to prevent it. PMID- 8727653 TI - AIDS: risk behaviors among rural Mexican women married to migrant workers in the United States. AB - International migration between Mexico and the United States has been acknowledged as a phenomenon that may contribute to the spread of AIDS in rural Mexico. The purpose of this study is to identify the information held by the participants regarding AIDS and to describe selected high-risk behaviors for AIDS transmission among a representative sample of rural women living in Mexico who are married to immigrant temporary workers to the United States. The women who participated in the study were married, of reproductive age, and had active sex lives with their spouses. Results revealed that most of the women interviewed had at least some knowledge about AIDS. Although some misconceptions were evident, most of the information they had was accurate. About one-third of the women felt at risk for AIDS, mostly because they doubted their husbands' fidelity, or because in the last five years they had donated blood, received a blood transfusion, or received an intramuscular or intravenous injection. The results of the study are discussed within the sociocultural context that surrounds the lives of the women interviewed. PMID- 8727654 TI - Rapid KABP survey for evaluation of NGO HIV/AIDS prevention projects. AB - The Johns Hopkins University HAPA Support Program (HSP) provided technical assistance to Save the Children (SC), a U.S.-based nongovernmental organization, to conduct a survey of knowledge, attitudes, beliefs, and practices (KABP) related to AIDS among rural Zimbabweans. The objectives of the HSP technical assistance were to field test a rapid KABP survey methodology and to assist SC to provide data that would contribute to their final project evaluation. The entire process of planning, implementation, preliminary data analysis, and preparation of a draft report of survey results was completed in a four-week period. A total of 660 respondents, aged 18-45 years, selected by a modified 30-cluster sampling method, were interviewed in two SC project areas. Although knowledge about HIV/AIDS was high, a number of misconceptions about HIV transmission and unfavorable attitudes to people with AIDS were noted. Of five knowledge and attitude variables that could be compared with the baseline survey results, 4 showed favorable changes and 1 showed an unfavorable trend. Comparing responses from those who were educated by SC with those who had other sources of information about HIV/AIDS, higher levels of knowledge were seen in the SC educated group and, in one area, somewhat greater willingness to care for family members with AIDS. However, there were no differences seen in other attitudes, beliefs, or in practices regarding condom use. The rapid KABP survey approach was successful in providing, with a relatively modest investment of resources, quantitative data useful for project evaluation, and for developing HIV/AIDS intervention strategies. PMID- 8727655 TI - Differences between Asian-American and white American dentists in attitudes toward treatment of HIV-positive patients. AB - In a survey of Asian (n = 115) versus white (Caucasian) (n = 920) dentists practicing in two boroughs of New York City, Asian dentists expressed significantly more negative attitudes toward and more unwillingness to treat HIV positive patients than did white dentists. Despite this consistent pattern across most survey items, the two groups were more similar regarding perceptions of professional obligation and their colleague's willingness to treat those with HIV. In an examination of the influence of acculturation processes on these attitudes, a comparison of attitude differences among the subgroup of Asian dentists receiving their dental education in the United States versus abroad showed some differences, with Asian dentists educated outside the United States expressing somewhat more negative attitudes. As Asian Americans become increasingly represented among practicing dentists in the United States, their relative unwillingness to treat HIV-positive patients may have an impact on access to oral health care among HIV-positive persons living in the United States. PMID- 8727656 TI - HIV testing among low-income African-American mothers. AB - HIV testing patterns were examined among low-income African Americans who were mothers of young school-age children. In-person interviews were conducted to determine whether African-American women had been tested for HIV; their sexual behaviors, including the number of sexual partners and condom use; and health care access and utilization. Forty-one percent of the women had been tested for HIV; 18 percent tested more than once. Levels of education, source of primary health care, and type of insurance were not associated with HIV testing. The total number of sexual partners for their current lifetime and within the past five years was significantly associated with their HIV testing status (chi 2 = 39.97; DF = 3; p < .01 and chi 2 = 66.68; DF = 3; p < .01 respectively). Women who have used condoms during their last intercourse (20%) were less likely to have been tested than women who did not use a condom (50%) (p < .01). Results suggest that low-income African-American women get tested for HIV. This suggests that these women understand the concept of risk and how certain behaviors may place them at risk. However, these women still confront conditions that place them at risk, such as having sex with multiple partners and/or partners with a history of incarceration, in addition to drug use. HIV testing may not serve as the most effective intervention for this sub-population of women. Educational and preventive measures should extend to women outside traditional high-risk populations by incorporating methods to promote protective behavioral changes which will empower women with self-esteem and confidence. PMID- 8727657 TI - A week in the country: a model respite program. AB - A family-oriented camping program for caregivers of children who are HIV infected is described. Major goals of the camping program consisted of: (1) providing respite care to mother and child, and, (2) training health care and mental health workers to provide special services, emphasizing the mental health needs of this population. PMID- 8727658 TI - [Plant acetyl-CoA carboxylases: strategy for survival]. PMID- 8727659 TI - [An aspect of the research on protein kinase C]. PMID- 8727660 TI - [PI3-kinase superfamily: structures and functions]. PMID- 8727661 TI - [Cdc2 activation by phosphorylation]. PMID- 8727662 TI - [Diversity of scavenger receptor molecules]. PMID- 8727663 TI - [Recent advance in the study of ascorbic acid biosynthesis]. PMID- 8727664 TI - [Ca2+/calmodulin dependent protein kinase cascade]. PMID- 8727665 TI - [Pathogenesis and therapy of non-steroidal anti-inflammatory drug (NSAID)-induced gastric mucosal injury]. PMID- 8727666 TI - [Meniscal repair]. PMID- 8727667 TI - [Perthes' disease]. PMID- 8727640 TI - Review of progress in sterol oxidations: 1987-1995. AB - Material dealing with the chemistry, biochemistry, and biological activities of oxysterols is reviewed for the period 1987-1995. Particular attention is paid to the presence of oxysterols in tissues and foods and to their physiological relevance. PMID- 8727668 TI - [Anterior decompression for cervical myelopathy caused by ossification of the posterior longitudinal ligament--anterior floating method of OPLL]. PMID- 8727669 TI - Selenoprotein gene expression during selenium-repletion of selenium-deficient rats. AB - Selenium repletion of selenium-deficient rats with 20 micrograms selenium / kg body weight as Na2SeO3 was used as a model to investigate the mechanisms that control the distribution of the trace element to specific selenoproteins in liver and thyroid. Cytosolic glutathione peroxidase (cGSHPx), phospholipid hydroperoxide glutathione peroxidase (PHGSHPx), and iodothyronine 5'-deiodinase (IDI) activities were all transiently increased in liver 16 to 32 h after ip injection with selenium. However, only cGSHPx and PHGSHPx activities increased in the thyroid where IDI activity was already increased by selenium deficiency. These responses were owing to synthesis of the seleoproteins on newly synthesised and/or existing mRNAs. The selenoprotein mRNAs in the thyroid gland were increased two- and threefold after the transitory increases in selenoprotein activity. In contrast, there were parallel changes in selenoprotein mRNAs and enzyme activities in the liver, with no prolonged rises in mRNA levels. The organ differences suggest that increased thryotrophin (TSH) concentrations, which are known to induce thyrodial IDI and mRNA, may control the mRNAs for all the thyroidal selenoproteins investigated and be a major mechanism for the preservation of thyroidal selenoproteins when selenium supplies are limited. PMID- 8727670 TI - Evidence for altered structure and impaired mitochondrial electron transport function in selenium deficiency. AB - Selenium (Se) deficiency in the experimental models, Coturnix coturnix japonica and Corcyra cephalonica, resulted in impaired mitochondrial substrate oxidations and lowered thiol levels. Studies with respiratory inhibitors confirmed reduced mitochondrial electron transport enzyme activities, especially at cytochrome c oxidase (COX), the terminal segment. Enhanced mitochondrial lipid peroxidation in Se deficiency was more pronounced in the heart tissue of the quail compared to other tissues. Glutathione peroxidase (GSH-Px) activity toward H2O2 and cumene hydroperoxide were generally low in the insect muscle tissue and activity toward H2O2 was maximal in the quail heart mitochondria that was not very sensitive to Se status. Lowered COX activity in Se deficiency was more directly correlated with the increased level of lipid peroxidation than with the GSH-Px activity measured, suggestive of Se mediated protective mechanisms independent of GSH-Px. Electron microscopic observations revealed structural changes such as loss of cristae with proliferative and degenerative changes of the mitochondria in Se deficiency. Involvement of Se in maintaining structure and functional efficiency of mitochondria is evident from the present study. PMID- 8727671 TI - Influence of season and diet on liver and kidney content of essential elements and heavy metals in Svalbard reindeer. AB - Samples of liver (n = 78) and kidney (n = 60) from Svalbard reindeer (Rangifer tarandus platyrhynchus Vrolik) collected at four different seasons in Svalbard were analysed for their content of Cd, Pb, Cu, Zn, Mn, and Se. The study shows that when animals are exposed to large seasonal variations in both the quality and quantity of food, it is crucial to relate element concentrations to the physiological condition of the animal, e.g., to look at seasonal fluctuations in the total element content of the different organs. PMID- 8727672 TI - The effect of manganese-induced hypercholesterolemia on learning in rats. AB - Since the exact mechanism of manganese (Mn)-induced learning disability is not known, we investigated the role of elevated cholesterol in rats exposed daily to 357 and 714 micrograms Mn/kg for 39 d. Significant Mn accumulation was accompanied by increased cholesterol content in the hippocampal region of Mn treated rats. The learning, which is based on the time needed to reach food placed at the exit of a T-maze after a 1-d training period, was significantly slower in exposed rats than in unexposed rats. The rats receiving 357 and 714 micrograms Mn/kg reached the food in 104.5 +/- 13.8 and 113.3 +/- 25.7 s, respectively, on d 30, whereas their untreated counterparts reached the food in 28.7 +/- 11.4 s. This delay was completely corrected to 29.3 +/- 7.8 and 30.7 +/- 6.0 s in rats with coadministration of an inhibitor of cholesterol biosynthesis with 357 and 714 micrograms/kg of Mn. The correction of impaired learning was associated with the normalization of hippocampal cholesterol, but the Mn level in this region of the brain was not influenced in rats treated with a drug that inhibits cholesterol biosynthesis. These results suggested that Mn-induced hypercholesterolemia is involved in Mn-dependent learning disability. PMID- 8727673 TI - Multielement analysis in serum of thyroid cancer patients before and after a surgical operation. AB - Concentrations of trace elements Zn, Cu, Fe, Mn, Mg, and Sr in the serum of 50 thyroid cancer patients and 50 healthy controls were analyzed comprehensively by Pattern Recognition Analysis Method. Based on the Principal Component Analysis Method, Zn, Cu, and Sr were found to be the principal elements with significant variations between patients and healthy controls. According to the Mahalanolis Distance Decision and Nonlinear Mapping Methods, the trace elements levels in the serum of cancerous patients were found significantly different from that of controls. For 16 thyroid malignant tumor patients who underwent surgery, sample points of patients after surgery were directional away from corresponding points before surgery. However, an overlapping was found and separation between two groups of points was not complete. This finding suggests that there may be a latency period for the restoration of trace element levels after removal of the malignant tumorous tissue. PMID- 8727674 TI - Determination of zinc fractions in human blood and seminal plasma by ultrafiltration and atomic absorption spectrophotometry. AB - A simple and reliable method is described which combines ultrafiltration technique with atomic absorption spectrophotometry to determine the Zn fractions in human blood plasma and seminal plasma. Ultrafiltrable, loosely bound, and firmly bound Zn can be measured using this method in the presence or absence of ethylenediaminetetraacetic acid (EDTA). The YMT membranes for the ultrafiltration must be rinsed thoroughly before use. In contrast to Zn in blood plasma, a large part of Zn in the seminal plasma was found to be ultrafiltrable. This method can be applied to study the physiologically active part of Zn in body fluids related to various disease states. PMID- 8727675 TI - Trace elements and lipid peroxidation in uremic patients on hemodialysis. AB - Trace elements and lipid peroxidation in 26 patients with chronic renal failure treated with hemodialysis and 25 healthy subjects were observed. Both plasma and erythrocyte trace elements and plasma malon dialdehyde (MDA) were examined immediately before and after hemodialysis. Increased levels of plasma Cu, MDA, and erythrocyte Pb, Mn, Zn, and a significantly decreased plasma Se, Zn, and erythrocyte Se were found in patients before hemodialysis. After a single hemodialysis, erythrocyte Mn, Cu, Zn, and plasma Cu, Al, and MDA were significantly increased whereas both plasma and erythrocyte Se were lower in patients than in healthy subjects. The level of MDA was not significantly changed during the single hemodialysis. Both plasma and erythrocyte Zn levels and plasma Cu and Al were significantly higher after hemodialysis than before hemodialysis. In conclusion, levels of trace elements are altered by hemodialysis, which may increase patient susceptibility to lipid peroxidation in uremia. PMID- 8727676 TI - Is erythrocyte alkaline phosphatase activity a marker of zinc status in humans? AB - The identification of an enzyme activity that responds to changes in Zn intake may serve as a useful biomarker for Zn status. Alkaline phosphatase (ALP) is a dimeric protein with each subunit containing two Zn atoms. The activity of ALP in erythrocytes (E) decreases as a result of a low Zn diet, which suggests that this enzyme may be a marker of Zn status. To investigate this further, we determined the response of E-ALP in six healthy subjects following supplementation with 50 mg Zn (4.2 x RDI) daily for 4 wk. A small but significant increase in plasma Zn was observed with supplementation (p < 0.05), whereas there was no significant change in E-Zn over the same period. Plasma and E-Cu showed no change. Conversely, the activity of E-ALP increased in all subjects from 1.7 +/- 0.5 to 5.9 +/- 0.7 U/g protein (mean +/- SE) (p < 0.0001). The small change observed in plasma Zn is not biologically significant in view of the many documented factors that influence its concentration. Our data support the hypothesis that E-ALP is a marker of Zn status in humans. PMID- 8727677 TI - A trace element preparation containing zinc increases the production of interleukin-6 in human monocytes and glial cells. AB - The in vitro effects of a trace element preparation (beres Drops Plus, BDP) on the biosynthesis of inflammatory cytokines interleukin (IL-6, IL-1, and tumor necrosis factor-alpha (TNF-alpha) were studied in human peripheral monocytes. The production of IL-6 was studied in a glioblastoma cell line, SKMG-4, as well. The trace element preparation BDP significantly stimulated both the constitutive and the endotoxin or IL-1 induced IL-6 production in monocytes or in glial cells, respectively, but revealed no or only modest effect on IL-1 and TNF-alpha production of monocytes. Moreover, BDP was able to reduce the inhibitory effect of a synthetic corticosteroid, dexamethasone on the biosynthesis of IL-6. The positive effect of the trace element preparation on the IL-6 production of monocytes from rheumatoid arthritis (RA) patients is comparable, to that of on the monocytes from healthy individuals, and similarly to healthy individuals was negligible on the IL-1 and TNF-alpha production. The detailed analysis of the composition of the preparation suggested, that the major active component in the stimulation of IL-6 production is Zn, but for the complete effect other trace elements are also required. PMID- 8727678 TI - Clinical case report: efficacy of yogic techniques in the treatment of obsessive compulsive disorders. AB - The aim of this study was to investigate the clinical efficacy of yogic techniques in the treatment of eight adults with obsessive-compulsive disorder (OCD). A specific yogic breathing pattern has been prescribed for the treatment of OCD, as well as others for treating generalized anxiety. A one year course of therapy was followed. Subjects improved on the Yale-Brown Obsessive-Compulsive Scale (Y-BOCS) comparing baseline with three, six, nine, & 12 month results (one way ANOVA for repeated measures, F(4,12) = 3.343, p < or = .046). Five patients completed the study (Y-BOCS results were 83%, 79%, 65%, 61% improvement, and one at-18%), group mean improvement of +54%. The Symptoms Checklist-90-R showed significant improvement comparing baseline and 12 months using two-tailed T-tests for OCD (t = 13.856, p < .001), anxiety (t = 3.167, p < .051), and global severity indexes (t = 7.314, p = .005). Perceived Stress Scale scores showed significant improvement for the five test periods (one-way ANOVA for repeated measures, F(4,12) = 9.114, p < or = .001). Five patients were well stabilized on fluoxetine prior to the study, three stopped medication after seven months or less, and two significantly reduced it, one by 25% and the other by 50%. These techniques, merit further study under controlled conditions and could help lead to new approaches for the treatment of OCD and perhaps other impulse control and anxiety-related disorders. PMID- 8727679 TI - Variations in sleep mentation as a function of time of night. AB - Mentation reports collected from sleep onset, Stage 2 and REM Stage awakenings, in the first part and in the second part of the night were analyzed both with systematic psycholinguistic and global measures. Results confirm the relationship between activation and the length of sleep mentation report shown by Antrobus. Length of the report increases with sleep time, but time does not modulate qualitative inter-stage differences. By partialling out the length of the report, many inter-stage differences disappeared; however significant differences remain in the global measure of bizarreness and in the psycholinguistic measure of visual imagery. These results cannot be explained entirely by differences in attention and memory and point to more basic differences in mental activity. PMID- 8727680 TI - EEG after prolonged mental activity. AB - The goal of the present study was to investigate the EEG spectral power changes caused by prolonged (6 hours) intensive mental loading consisted of calculation and choice-reaction tasks. The EEGs were analysed both during the tasks performance and in rest periods at the beginning of the work (after subjects had already mastered the tasks) and at the end. The significant increase both of slow (delta, theta) and fast (beta) activities was shown at the end of the work. The observed EEG spectral power changes with time on task were similar during the different tasks performance as well as in the rest conditions indicating the general character of the observed phenomena. These results are interpreted as reflecting the deterioration of general brain state activity caused by prolonged mental work and the simultaneous additional brain activation to provide sufficient vigilance level and successful tasks performance (the subjects being highly motivated). PMID- 8727681 TI - Effect of repeated visual traumatic stimuli on the event related P3 brain potential in post-traumatic stress disorder. AB - Post-Traumatic Stress Disorder (PTSD) patients are characterized by a hypersensitivity to traumatic stimuli which may be expressed as an automatic and involuntary cognitive response. Electrophysiologically this can be recorded as an augmented visual P3 (P300) event related potential (ERP). This study examined P3 changes in response to repeated traumatic pictorial stimuli presented in the form of a visual discrimination oddball paradigm to 40 Israeli combat veterans with and without PTSD. Subjects were asked to press a button when target stimuli (domestic animal pictures) appeared, and to ignore all non-target stimuli (irrelevant pictures of furnishings/flowers and traumatic combat related pictures). On average, P3 in response to combat related pictures was earlier and approximately 5 times greater in amplitude for the PTSD patients as compared to the controls. Repeated combat related pictures stimuli presentation resulted in a rapid and appreciable P3 amplitude reduction and latency prolongation. This effect was not observed for the target stimuli. These findings suggest that a gradually reduced amount of attentional resource is required and allocated to the processing of repeated CRP stimuli. This may occur as a consequence of the activation of an inhibitory mechanism related to the cognitive processing of traumatic stimuli. PMID- 8727682 TI - Auditory event-related potentials while ignoring tone stimuli: attentional differences reflected in stimulus intensity and latency responses in low and highly hypnotizable persons. AB - Effect of hypnotic susceptibility level on auditory event-related potentials (AERPs) was studied as subjects were instructed to ignore tones while reading a novel or counting one's pulse. Assessed previously on two hypnotic susceptibility scales [Harvard Group Scale of Hypnotic Susceptibility; Stanford Hypnotic Susceptibility Scale, Form C (SHSSC)], subjects were 12 low (lows; SHSSC 0-2) and 12 highly (highs; SHSSC 9-12) hypnotizable, right-handed college students. AERPs were recorded at C3, C4, and Cz to 50 ms 1.961 tone pips 50, 60, 70 and 80 dB intensities, pseudorandomly presented at 1.5 intervals. As predicted, highs had significantly smaller N1 and P2 amplitudes than did lows when ignoring tones. As stimuli intensities increased, N1 latencies decreased for lows while N1 latencies increased for highs. N1 latency slopes across the 50 to 80 dB intensities were significantly more negative for lows than highs; slopes correlated significantly with both hypnotizability and absorption (Tellegen Absorption Scale). Thus, the highs appeared to divert greater attentional processing to the tasks at hand, particularly as the tones increased in intensity, and were slower to respond to not-to-be-attended stimuli. These results are interpreted as further evidence for hypnotic susceptibility being associated with efficient attentional processing such that highs can more effectively partition attention towards relevant stimuli and away from irrelevant stimuli than can low hypnotizables. PMID- 8727683 TI - The endogenous eyeblink and hypnotic susceptibility in a real-simulator design. AB - This study investigated the relationship between hypnotic susceptibility and the endogenous eyeblink with 27 subjects who were assigned to groups of high susceptibles, low susceptibles and simulators on the basis of cutoff scores from the Harvard Group Scales of Hypnotic Susceptibility: Form A and the Stanford Hypnotic Susceptibility Scales: Form C. Using a repeated-measures design, oculomotor data were collected during two separate conditions, waking and hypnotized, while subjects performed a visual task requiring the discrimination of short light flashes (200 ms) from long light flashes (400 ms). Although results partially replicated previous studies, with high susceptibles blinking significantly less than low susceptibles across both conditions, no effect was found for the hypnotic state. Failure of the simulating group to meet assumptions in the waking condition allowed no conclusions regarding impact of task demands on the endogenous eyeblink. PMID- 8727684 TI - Weak electromagnetic fields increase the amplitude of the pattern reversal VEP response in patients with multiple sclerosis. AB - Visual evoked potential (VEP) studies are widely used for the diagnosis of multiple sclerosis (MS) and are also useful in monitoring the effects of various therapeutic modalities in the disease. Brief, extracerebral applications of picotesla (pT) range flux intensity electromagnetic fields (EMFs) of low frequency have been shown efficacious in the treatment of motor and cognitive symptoms in MS implying that this treatment modality improves action potential transmission in demyelinating pathways. This report documents three MS patients with a remitting-progressive course in whom two successive brief extracerebral applications of pT range EMFs caused an immediate increase (and normalization) of the amplitudes of the visual evoked response in the eye previously affected by optic neuritis. However, the pretreatment prolonged latencies of the evoked responses remained essentially unchanged after the administration of EMFs. Since the latency of the VEP reflects the degree of conduction velocity and the amplitude the degree of conduction block in demyelinating optic pathways, the report demonstrates that extracerebral applications of these EMFs may rapidly reverse conduction block in demyelinating fibers. Reversal of the conduction block, which is though to be related to changes in axonal Na+ and K+ channels and synaptic neurotransmitter release, accounts for the immediate improvement of vision and other neurological deficits observed in MS patients following exposure to these EMFs. PMID- 8727685 TI - Bidirectional effect of electromagnetic fields on ketanserin-induced yawning in patients with multiple sclerosis: the role of melatonin. AB - 5-HT2 receptors regulate sleep including yawning behavior. Ritanserin, a selective 5-HT2A receptor antagonist, increases the duration of slow wave in rats and humans. This effect is more pronounced during the light period when melatonin plasma levels are low; melatonin inhibits the sleep effects of ritanserin. These findings indicate that melatonin co-determines the effects of ritanserin on sleep. In a cohort of multiple sclerosis (MS) patients ketanserin, a selective 5 HT2A receptor antagonist, induces recurrent yawning particularly when administered in daytime. The frequency of yawning induced by the drug was modified by AC pulsed picotesla flux electromagnetic fields (EMFs) which affect melatonin secretion. Two MS patients are presented in whom the frequency of ketanserin-induced yawning was altered in opposite directions by these EMFs. The first patient, a 50 year old woman with a remitting-relapsing course, developed recurrent yawning and sleepiness after administration of ketanserin (10 mg, PO). Yawning was decreased dramatically during application of EMFs but was unaffected by a placebo EMFs treatment. The second patient, a 35 year old man with a chronic progressive course, manifested a single and brief yawn after administration of an equal dose of ketanserin. Yawning was increased dramatically during application of EMFs while remaining unchanged during a placebo EMFs treatment. These observations demonstrate a bidirectional effect of picotesla flux EMFs on ketanserin-induced yawning which may be related to differences in daytime melatonin plasma levels among MS patients. If validated by estimations of melatonin plasma levels in a larger cohort of patients the information derived from the effects of picotesla EMFs on ketanserin-induced yawning could be used to: (a) assess pineal melatonin functions in patients with MS; (b) indicate differences in pineal functions between male and female MS patients; and (c) indicate a relationship between plasma melatonin levels and the fatigue of MS. PMID- 8727686 TI - Application of weak electromagnetic fields facilitates sensory-motor integration in patients with multiple sclerosis. AB - Electrophysiological studies in behaving animals have shown the function of cerebral serotonin (5-HT) neurons to be altered in association with motor output in both the tonic and repetitive modes and also in relation to an orienting response. Brainstem 5-HT neurons increase their firing rate two to five-fold during repetitive motor activity to facilitate motor output while simultaneously suppressing transmission in sensory pathways. Reciprocally, during an orienting response motor activity is suppressed and 5-HT neuronal activity is inhibited to facilitate transmission of sensory information. These reciprocal changes in 5-HT neuronal activity serve to facilitate brainstem reticular sensory-motor integration which, due to 5-HT neurotransmission deficiency, may be disrupted in patients with multiple sclerosis (MS). For instance, MS patients are unable to process auditory information in the presence of competing ambient stimuli, while under a controlled laboratory environment they demonstrate unimpaired verbal information processing. This report concerns three MS patients who experienced rapid deterioration in balance resulting in falling when subjected, during ambulation, to distracting external auditory stimuli. After receiving a series of treatments with low frequency picotesla range intensity electromagnetic fields (EMFs), which were applied extracranially for brief periods, these patients experienced resolution of these symptoms with ambulation being unaffected by auditory stimuli. It is suggested that application of picotesla EMFs may restore abnormal reticular sensory-motor integration in MS patients with the effect being related to facilitation of 5-HT neurotransmission at both junctional (synaptic) and nonjunctional neuronal target sites. PMID- 8727688 TI - Weak electromagnetic fields potentiate the effects of 4-aminopyridine in multiple sclerosis. PMID- 8727687 TI - Freezing of gait in Parkinson's disease is improved by treatment with weak electromagnetic fields. AB - Freezing, a symptom characterized by difficulty in the initiation and smooth pursuit of repetitive movements, is a unique and well known clinical feature of Parkinson's disease (PD). It usually occurs in patients with long duration and advanced stage of the disease and is a major cause of disability often resulting in falling. In PD patients freezing manifests most commonly as a sudden attack of immobility usually experienced during walking, attempts to turn while walking, or while approaching a destination. Less commonly it is expressed as arrest of speech or handwriting. The pathophysiology of Parkinsonian freezing, which is considered a distinct clinical feature independent of akinesia, is poorly understood and is believed to involve abnormalities in dopamine and norepinephrine neurotransmission in critical motor control areas including the frontal lobe, basal ganglia, locus coeruleus and spinal cord. In general, freezing is resistant to pharmacological therapy although in some patients reduction or increase in levodopa dose may improve this symptom. Three medicated PD patients exhibiting disabling episodes of freezing of gait are presented in whom brief, extracerebral applications of pulsed electromagnetic fields (EMFs) in the picotesla range improved freezing. Two patients had freezing both during "on" and "off" periods while the third patient experienced random episodes of freezing throughout the course of the day. The effect of each EMFs treatment lasted several days after which time freezing gradually reappeared, initially in association with "off" periods. These findings suggest that the neurochemical mechanisms underlying the development of freezing are sensitive to the effects of EMFs, which are believed to improve freezing primarily through the facilitation of serotonin (5-HT) neurotransmission at both junctional (synaptic) and nonjunctional neuronal target sites. PMID- 8727689 TI - Effects of ceruletide and haloperidol on auditory evoked potentials in the cat brain. AB - The influence of cholecystokinin-like peptide, ceruletide, on EEG and auditory evoked potentials (AEPs) was studied in nine cats. The cats were bearing electrodes implanted in the auditory cortex, hippocampus, reticular formation and cerebellum. Reference drugs used were haloperidol and neostigmine. The hippocampus showed the strongest effect of ceruletide, whereas the cerebellum was virtually unresponsive. The amplitude of AEPs was increased by peptide, an effect lasting up to 21 days which, according to amplitude frequency analysis (AFC) was due to an augmented theta response. The latter possibly indicates increased signal transfer to, or through, the brain structure in question, particularly in the hippocampal neurons. The effects of haloperidol and neostigmine did not reflect those of ceruletide and lasted only a few hours. PMID- 8727690 TI - Effects of intravenous L-dopa on P300 and regional cerebral blood flow in parkinsonism. AB - The P300 and regional cerebral blood flow were measured before and after intravenous injection of L-dopa in 10 patients with idiopathic Parkinson's disease and 10 patients with vascular parkinsonism. The P300 was measured with an evoked potential recorder using an oddball paradigm and the regional cerebral blood flow was measured using the stable xenon computed tomography method. The P300 latency was significantly longer and the regional cerebral blood flow in the cerebral cortex and basal ganglia was significantly lower in the Parkinson's disease group and the vascular parkinsonism group than in the age-matched healthy control group. The intravenous injection of L-dopa improved these abnormalities significantly in the Parkinson's disease group but did not improve these abnormalities in the vascular parkinsonism group. Cognitive function is considered to be impaired in Parkinson's disease and vascular parkinsonism and L dopa is considered to improve it in Parkinson's disease. PMID- 8727691 TI - Evoked brain rhythms are altered markedly in middle-aged subjects: single-sweep analysis. AB - In this study, a new method for analysis of single frequency responses was applied to compare auditory evoked potentials between young (18-30 years old) and middle-aged (50-55 years old) adults. The method was related to the evoked brain rhythmicities concept and permitted to quantify separately the magnitude, phase locking, and enhancement against prestimulus activity of single sweeps. Thus, a constellation of three single-sweep parameters was used. No differences could be detected between the two age groups in the conventionally averaged potentials. However, significant age-related alterations were found at the level of single sweep analysis: 1) At the three midline locations (Fz, Cz, and Pz) older adults manifested theta phase-locking and enhancement that were significantly stronger than those in young adults; 2) In contrast, the alpha responses only over the frontal brain area were stronger in phase-locking and enhancement in middle-aged than in young adults, which implies that the alpha response system may be specifically related to frontal brain functioning during aging. PMID- 8727692 TI - Sound stimulation-induced vasomotor reflex in the central artery of the rabbit ear. AB - Effects of sound stimulation on the central artery of the rabbit ear were studied as a somato-autonomic reflex. Vasoconstriction and dilatation, caused by metronome sound stimulation, were estimated from the temperature fluctuations in the central artery of the ear, measured by a thermistor. To enhance the detection of temperature rises, moderately high background levels of arterial tone were established by exposing the tips of the ears to water at a temperature of 10 degrees C or 5 degrees C, prior to sound stimulation. A fall in arterial temperature due to vasoconstriction was observed immediately after the start of the 1-min sound stimulation, with a subsequent temperature rise which overshot the original basal level due to vasodilatation. A positive correlation between the ear temperature before sound stimulation and the temperature fall (p < 0.01), and a negative correlation between the ear temperature and the temperature rise (p < 0.05) were obtained. The temperature fall was blocked by phenoxybenzamine (9 mg/kg, i.p., p < 0.01). The subsequent rise was not influenced by atropine (3 mg/kg, i.p.) or phenoxybenzamine, however, it was attenuated by hexamethonium (6 mg/kg, i.p., p < 0.05). The temperature fall at the beginning of sound stimulation was related to alpha-adrenergic mechanism. The subsequent temperature rise was thought to be related to parasympathetic mechanism, excluding cholinergic mechanism. PMID- 8727693 TI - A new in vitro model of specific targeting therapy of cancer: retargeting of PWM LAK cells with bispecific antibodies greatly enhances cytotoxicity to hepatocellular carcinoma. AB - For the purpose of establishing a new in vitro model of adoptive immunotherapy, we synthesized two kinds of bispecific antibodies (BsAbs), i.e., (OK x L) BsAbs constructed with both OKT-3 (anti-CD3) and L-7-6 (anti-HCC), and (3G x L) BsAbs constructed with 3-G-8 (anti-CD16) and L-7-6 antibodies. These two BsAbs, having pairs of binding arms on their single molecule, showed similar binding to target cells as the parental monoclonal antibodies (OKT-3, 3-G-8 and L-7-6), when examined with FACS. Newly devised in vitro cytotoxicity tests revealed that LAK or PWM-stimulated LAK (PWM-LAK) cells did not show any significant cytotoxic activity to HCC cells, while both effector cells equally showed greatly enhanced cytotoxicity to HCC even at a low effector/target (0.3) in the presence of BsAbs (OK x L) for the efficient retargeting of the effector cells. Inasmuch as PWM-LAK cells proliferate in vitro 3-5 times faster than LAK cells, adoptive immunotherapy using PWM-LAK cells in combination with (OK x L) BsAbs should be very promising. PMID- 8727694 TI - Analysis of plasma metabolites during human PET-studies with three receptor ligands, [11C]YM-09151-2, [11C]doxepin and [11C]pyrilamine. AB - Carbon-11 labeled metabolites in human plasma were analyzed by high-performance liquid chromatography during positron emission tomography (PET) studies using the dopamine D2 ligand [11C]YM-09151-2 as well as the histamine H1 ligands [11C]doxepin and [11C]pyrilamine. For all the three tracers, blood clearance of the radioactivity was extremely rapid after an i.v. injection. The plasma protein binding of [11C]YM-09151-2 and [11C]doxepin had protective effects upon the metabolic alteration of the ligands, whereas [11C]pyrilamine was free from the protein-binding and immediately degraded. The degradation of [11C]doxepin was more rapid in epileptic patients on medication than in normal subjects. These results indicate that analysis of metabolites in the plasma is necessary to determine the accurate arterial input function for quantitative PET measurement. PMID- 8727696 TI - Chronic bile diversion to the urinary bladder enhances cholecystokinin release and suppresses gastric inhibitory polypeptide release in dogs. AB - The role of intestinal luminal bile on cholecystokinin (CCK) and gastric inhibitory polypeptide (GIP) release was examined in dogs with chronic external bile diversion. In 6 mongrel dogs, cholecysto-jejuno-cystostomy (C-J-C) was performed using a small segment of the middle small intestine interposed between the gallbladder and urinary bladder with section of the common bile duct. Butter dissolved in 30 ml of lukewarm water was orally ingested before surgery, and same amount of butter solution with or without graded volumes of canine bile were orally ingested between four to six weeks after C-J-C surgery. Increases in both plasma triglyceride and GIP levels after butter ingestion were almost completely abolished by C-J-C, but they were restored by oral bile feeding in a volume dependent manner. Both basal and fat-stimulated CCK release were enhanced significantly by C-J-C, and oral bile feeding inhibited fat-stimulated CCK release by bile in a dose dependent manner. These data suggest that intraluminal bile regulates basal and fat-stimulated CCK release and fat-stimulated GIP release in dogs. PMID- 8727695 TI - PMA and ionomycin differently affect atrial natriuretic peptide stimulated cyclic GMP production in rat mesangial cells. AB - How 4 beta-phorbol 12-myristate 13-acetate (PMA) and ionomycin (Io), a calcium ionophore, affect on the atrial natriuretic peptide (ANP) stimulated cyclic-3',5' guanosine monophosphate (cGMP) production in cultured rat mesangial cells was examined. Cultured mesangial cells were prepared by isolated glomeruli from Sprague Dawley rats employing the sieving method and were used between the 3rd and 15th passage for experiments. cGMP and protein contents were measured by radioimmunoassay and Lowry method. Incubations with effectors were carried out either in the presence or absence of 0.5 mM 1-methyl-3-isobutyl-xanthine (MIX). The intracellular concentration of calcium ([Ca2+]i) was determined by using the Fura-2 method. Pretreatment with PMA, an activator of protein kinase C (PKC), attenuated ANP stimulated cGMP production in a time- and dose-dependent fashion, while alpha PDD (an inactive analog of PMA) did not inhibit cGMP production. PMA inhibition was reversed by addition of staurosporine, a protein kinase C inhibitor. Io attenuated ANP stimulated cGMP production in the absence but not in the presence of MIX. These findings suggested that PMA acts on ANP receptor or guanylate cyclase via activation of PKC in rat mesangial cells. Io may inhibit ANP stimulated cGMP production via activation of cyclic nucleotide phosphodiesterase. PMID- 8727697 TI - Enzymes in intestinal juice from patients with liver diseases and colon polyps: measurement of bilirubin, alkaline phosphatase, aspartate aminotransferase and lactate dehydrogenase. AB - Since the amounts of hepatogenous enzymes discharged into the intestinal tract remain unknown, this study was initiated to evaluate the amounts of the enzymes in the intestinal tract. Whole gut lavage fluid (polyethyleneglycol electrolyte solution) was administered orally to 42 subjects, consisting of 5 patients with hepatoma, 10 with chronic hepatitis, 10 with colon polyps, and 17 control subjects without liver disease. Two hr after the large intestinal lavage, the digestive tract juice was aspirated by colonoscopy, and the bilirubin (Bil), alkaline phosphatase (ALP), aspartate aminotransferase (AST), and lactate dehydrogenase (LDH) in the aspirates were measured. A positive correlation between the AST and LDH values was found, and a significant difference in these values between the hepatic disorders and the normal controls was noticed. A significant positive correlation between the ALP and Bil values was found, and a statistical difference in these values between the group of colon polyps and the controls and other groups was observed. This lavage fluid technique enables to estimate the amounts of hepatic enzymes discharged into the intestinal tract, thereby opening a new avenue for future enzyme research. PMID- 8727698 TI - Correlation analysis between bladder cancer and cigarette smoking in various countries. AB - Cigarette smoking is considered to be a risk factor of bladder cancer. We investigated the relationship between bladder cancer mortality rates and cigarette smoking prevalence in various countries, and examined whether only the difference of smoking prevalence could explain the difference of mortality rates of bladder cancer in each country or not. The age-adjusted mortality rates from bladder cancer were high in European countries and low in Asian countries. The sex ratios ranged between 2 to 6 in most of the countries. For 20 countries, the mortality rate from bladder cancer was not correlated with the cigarette smoking prevalence. However, when the analysis was restricted to 16 countries in Europe, North America and Oceania (Caucasians), the correlation coefficients between them were high, being 0.68 (p < 0.01) in males and 0.49 (0.05 < p < 0.1) in females. In both Japan and the United States, cigarette smoking prevalence declined while the incidence of bladder cancer increased in males, both being stable in females. Race might be considered to be a strong risk factor of bladder cancer, followed by the prevalence of cigarette smoking. It is necessary to follow the changes of incidence rates of bladder cancer in the future to evaluate the influence of cigarette smoking. PMID- 8727699 TI - Changes in the distribution of nerve conduction velocities in diabetics. AB - To examine which of the faster or slower large myelinated nerve fibers (alpha fiber group) are affected earlier by diabetes mellitus (DM), the distribution of nerve conduction velocities (DCV) and conventional sensory nerve conduction velocities (SCV) in the median nerve were measured in 10 male DM patients aged 35 58 (mean 50) years. The patients had suffered from DM for 0-30 (mean 11) years; their fasting blood sugar (FBS) levels were 105-363 (mean 219) mg/100 ml. The DCV was expressed by the conduction velocities below which 10, 20, ......, 80 and 90% of active fibers lie (V10, V20, ......, V80 and V90 velocities, respectively). In DM patients, the V30 to V90 velocities of the DCV as well as the SCV were significantly smaller than those of age-matched male controls. The results of multiple regression analysis indicated that FBS was inversely related to V50 to V90 velocities; duration of the disease was also significantly related to V80 and V90 velocities. These data suggest that faster nerve fibers are more susceptible to DM than slower fibers are; changes in conduction velocities of the faster fibers are closely associated with FBS and duration of DM. PMID- 8727700 TI - Isolation and characterization of two monoclonal antibodies that recognize different epitopes of the human c-kit receptor. AB - After immunizing mice with a human megakaryoblastic leukemia cell line, M-MOK, we obtained two monoclonal antibodies which recognize the human c-kit receptor. The monoclonal antibodies, designated MTK1 and MTK2, were found to specifically recognize Balb/3T3 cells transfected with human c-kit cDNA and not parent Balb/3T3 cells while showing different immunological, biochemical and biological behaviors. Both allowed visualization of the 140 kDa c-kit protein by Western blot analysis, but MTK1 detected only positive band with non-reducing conditions for sodium dodecyl sulfate-polyacrylamide gel electrophoresis. MTK1 partially inhibited the stem cell factor (SCF) induced proliferation of M-MOK cells, whereas, MTK2 was without effect. MTK1 also inhibited the bone marrow derived colony forming unit granulocyte/macrophage (CFU-GM) formed by granulocyte macrophage colony stimulating factor (GM-CSF) and SCF. Not only anti-CD34 antibodies (HPCA-1) but also MTK1 could be shown to concentrate bone marrow CFU GM and burst forming unit erythroid (BFU-E) effectively. The presently described monoclonal antibodies may therefore be useful for functional analysis of the ligand binding domain of the human c-kit receptor, as well as for further classification of hematopoietic stem cells in addition to the CD34 positive cells. PMID- 8727701 TI - Flow cytometric DNA analysis of abnormal endometrium. AB - Samples of abnormal endometrium, which were composed of 5 cases of atypical hyperplasia (ATH), 12 cases of well differentiated adenocarcinoma (G1), 17 cases of moderately differentiated adenocarcinoma (G2), 11 cases of poorly differentiated adenocarcinoma (G3) and 2 cases of serous papillary adenocarcinoma (SPA), were analyzed by flow cytometry. With respect to the ploidy level, all cases of both ATH and G1 showed diploidy, and the proliferative index (PI) was 14.32 and 16.33, respectively. In contrast, 3 of G2 cases, 3 of G3 cases and all of SPA cases showed aneuploid patterns, and the PI was 21.06, 27.91 and 31.52, respectively. There were statistical differences between the former group and the latter group as to both the ploidy and the PI. It was found that ATH and G1 had similar biological behavior. PMID- 8727702 TI - Characterization of serum anti-phospholipid antibodies in patients with multiple sclerosis. AB - We measured serum antiphospholipid antibodies (aPL) in patients with multiple sclerosis (MS) using enzyme linked immunosorbent assay (ELISA) and examined the correlations between these antibodies and MS. This study included thirty-two patients with clinically definite MS, thirteen patients with other autoimmune neurological diseases excluding collagen diseases (disease control A), eight patients with collagen vascular diseases (disease control B) and twenty-six healthy persons (normal control). In MS group IgG antibody against cardiolipin (CL) was detected in 3 (9%); among them, cofactor (beta 2-glycoprotein I) dependency was shown in 2 but one was cofactor independent. IgM antibody was elevated in 14 of 32 patients (44%) with MS, but cofactor dependency was not determined. However, this was significantly higher in frequency than that of the disease control A (p < 0.01) and normal control (p < 0.01). Results of antibodies against phosphatidylserine were found similar to CL, but antibodies against phosphatidylcholine were in most cases negative. Each of anti-CL IgG antibody purified from four patients with diverse immunological disorders (primary antiphospholipid antibody syndrome, MS, polyarteritis nodosa and systemic lupus erythematosus) had different reactivities against DNA. In addition, the aPL positive group in MS possessed the autoantibodies such as antinuclear antibody at higher rate than the negative group. However, clinically two groups of MS were indistinguishable. The higher incidence of aPL may imply that a broad spectrum of autoantibodies might be produced in MS; some antibodies presumably related directly to MS pathogenesis are yet to be identified. PMID- 8727703 TI - Simple method for quantifying disorientation of myofiber in endomyocardial biopsy specimens using an image analyzer. AB - The disarray of cardiac myofibers was morphometrically evaluated in biopsy specimens, obtained from patients with hypertrophic cardiomyopathy (HCM), hypertensive heart disease (HHD), chronic phase myocarditis and controls. Microphotographs of myocardium were taken at a final magnification of x 250. For each segment of myofibers, the longitudinal direction was traced on a transparent sheet, and the angle of the traced direction to the baseline was measured by an image analyzer. The standard deviation of the angles was used as an indicator of myofiber disarray. The histograms showed a narrow variation in the control group, but a wide variation in the patients with HCM, HHD and chronic phase myocarditis. The mean value of standard deviations of myofiber angles in HCM was significantly larger than that in the other groups. In HCM, the standard deviation of the myofiber angle proved to correlate positively with IVST/LVPWT (the thickness ratio of interventricular septum to the left ventricular posterior wall) (r = 0.70, p < 0.05), and also with LVEDP (left ventricular end-diastolic pressure) (r = 0.66, p < 0.05). In conclusion, the image analyzer serves as a simple and useful tool in quantifying the disorientation of myofibers and estimating the correlation between the histological and clinical findings. PMID- 8727704 TI - Indications for laparoscopic surgery of ovarian tumors. AB - To investigate the criteria for laparoscopic surgery in ovarian tumors, a retrospective review of indications, preoperative assessments using transvaginal ultrasonography, serum CA 125, surgical methods and diagnosis and post-operative complications were analyzed. Forty-three patients underwent surgical treatment for ovarian tumors, from November 1992 to October 1993. Of the 43 patients with ovarian tumors, 19 (44%) underwent laparoscopic surgery, and 24 (56%) underwent laparotomy. Laparoscopic surgery was performed in 18 of 34 (53%) patients with benign ovarian cysts. Eight patients with malignant ovarian tumors were treated by laparotomy. Coincidence of preoperative assessment and surgical diagnosis was 95%. No major complications were observed in the laparoscopic surgery. We suggest that malignant ovarian tumors and inextricable adhesions are contraindications to laparoscopic surgery. We propose the following criteria for laparoscopic surgery by classification into three groups: A) good indication: benign cystic tumors with good mobility, B) deliberation indication: benign tumors with poor mobility, including some dermoid cysts and endometrial cysts, C) contraindication: malignant tumors and dense adhesions. We conclude that indications of laparoscopic surgery for ovarian tumors should be determined by careful preoperative assessment using transvaginal ultrasonography and tumor markers. PMID- 8727705 TI - The differences in the expressions of visual pigments and transducin in photoreceptor cell differentiation. AB - The distribution and accumulation of visual pigments, i.e., rod pigment, rhodopsin and red sensitive cone pigment, iodopsin, and transducin in the retina of chicken and chicken embryo were investigated immunohistochemically using their specific antibodies. The immunoreactivities of these proteins appeared at the early stage of photoreceptor differentiation (embryonal day 15) and increased in the photoreceptor cells appeared to reach maximum at the end of the embryonal period (embryonal day 20). On the other hand, although the immunoreactivity of beta gamma subunit of transducin (T beta gamma) was detected at embryonal day 15, the expression level of T beta gamma still remained in low level during the embryonal period. These observations suggest that both T beta gamma and visual pigments are expressed during the embryonic period in chicken photoreceptor cells, but their accumulations in the cells are different. PMID- 8727706 TI - Metallothionein synthesis induced by interferon alpha/beta in mice of various zinc status. AB - We studied the ability of interferon alpha/beta (IFN) to induce metallothionein (MT) synthesis in mice. Male mice were injected intraperitoneally with mouse IFN (5 x 10(5) IU/mouse). Plasma Zn levels were reduced at 4 hr after injection, reached a minimum value at 6 hr, and then returned to the control level at 8 hr. Hepatic MT concentrations began to increase at 4 hr and reached maximum values at 6 hr. Induction of MT gene expression and protein synthesis was confirmed by Northern blot analysis and radioimmunoassay, respectively. The induction of MT synthesis in the liver by IFN was dose-dependent. The data suggest that induction of MT-mRNA and the protein in the liver by IFN occurs rapidly but is rather transient. Furthermore, MT synthesis was not induced by IFN in the liver of mice given a Zn-deficient diet, whereas IFN induced increases in the activity of 2',5' oligoadenylate synthetase in the spleen were unaffected by Zn status. Thus, induction of hepatic MT synthesis by IFN is influenced by Zn status. PMID- 8727707 TI - Changes in viscoelasticity of the myocardium during cardioplegic arrest. AB - To evaluate the efficacy of myocardial preservation during open heart surgery, we measured the viscoelasticity of the canine myocardium during cardioplegic arrest. A transfer function method was used for the measurement with a monitoring system consisting of a vibrator, a function generator, accerometers and a signal processor. Six mongrel dogs were put on cardiopulmonary bypass and after measurement of control hemodynamics, they were subjected to cardioplegic arrest at myocardial temperatures ranging from 4 to 32 degrees C. Viscoelasticity was measured at every 15 min and the cardioplegic solution was added every 30 min. After two hr of cardioplegic arrest, the myocardium was reperfused and postischemic hemodynamics were measured after 30 min of non-working beating. Satisfactory myocardial function returned in 3 hearts with the myocardial temperatures below 24 degrees C with myocardial viscoelasticity within the control range. Moderately decreased myocardial contractility was noted in a heart kept at temperature of 27 degrees C and its viscoelasticity remained in the control range of 90 min of ischemia and then began to decrease. In 2 hearts kept at temperatures higher than 29 degrees C, severely depressed myocardial contractility was noted, and viscoelasticity decreased transiently at 45 to 60 min and then returned to control levels. These results suggested usefulness of continuous monitoring of the viscoelasticity in early detection of its degenerative alterations due to impaired myocardial preservation during open heart surgery. PMID- 8727708 TI - Characterization of protein kinase C expression in human kidney. AB - Protein kinase C (PKC), the calcium and phospholipid-dependent kinase, is a key enzyme for regulation of growth and differentiation of various cells and tissues. Abnormalities of PKC modulation may cause diseases such as neoplasia. The present study was designed to demonstrate localization of PKC in normal human nephron segments. Surgically extirpated specimens for renal tumor were used. PKC was determined by immunoblotting and immunohistochemical methods using specific antibody against PKC that recognizes alpha and beta subtypes of this enzyme. Immunoblotting study demonstrated PKC in the cytosolic fractions of homogenized cortical tubules and medullary tubules prepared from morphologically normal part of the kidney. PKC was faintly detected in glomerular supernatants. Immunohistochemical study revealed that PKC was present throughout the nephron and especially concentrated in the cytoplasm of the proximal tubules and the medullary collecting ducts. The results indicate that PKC appears to play a role in regulation of normal nephron function in human kidney. PMID- 8727709 TI - Capacity of H2O2 release from monocytes in steroid-sensitive nephrotic syndrome. AB - The pathogenesis of steroid-sensitive nephrotic syndrome (SSNS) is poorly defined. We previously demonstrated that monocytes from SSNS patients with proteinuria were activated to display exaggerated phagocytosis of opsonized particles and paradoxically reduced chemotaxis. In this study, we evaluated the capacity of hydrogen peroxide (H2O2) release from monocytes in 19 patients with SSNS and 13 healthy controls, by exposure to phorbol myristate acetate (PMA), using scopoletin method. Of 19 patients of SSNS, 7 were proteinuric and 12 in remission. The H2O2 release was significantly higher in SSNS patients with proteinuria than those in remission or normal controls [177.49 +/- 94.75 (mean +/ S.D.) vs. 60.67 +/- 58.89 (p < 0.02) or 85.02 +/- 48.62 nmol/90 min/mg cell protein (p < 0.05)]. Follow-up measurements in two SSNS patients showed that H2O2 release was reduced when proteinuric condition was improved to be in remission. Our data suggest that monocytes in SSNS with proteinuria were activated and were prepared to receive some extracellular signaling leading to protein kinase-C activation for releasing H2O2. PMID- 8727710 TI - Effects of OP 2507, a stable analogue of prostaglandin I2, on carbon tetrachloride-induced liver damage in starved rats. AB - It has been reported that vasodilatory prostaglandins have cytoprotective effects against various types of liver damage. We investigated the effects OP 2507, a stable analogue of prostaglandin I2, on carbon tetrachloride-induced liver damage in starved rats. Intraperitoneal administration of OP 2507 at 1,500 micrograms/kg lessened both an increase in serum alanine aminotransferase activity and an inhibition of starvation ketosis, both of which were induced by carbon tetrachloride. At lower doses, however, OP 2507 not only failed to ameliorate the carbon tetrachloride-induced changes, but it actually exaggerated them. Although the deterioration of carbon tetrachloride-induced liver damage by lower doses of OP 2507 was not statistically significant, it seems possible that OP 2507 has dual effects on carbon tetrachloride-induced liver damage. While none of the three agents cimetidine, reduced glutathione and deferoxamine, prevented increase in serum alanine aminotransferase activity induced with lower dose OP 2507, allopurinol had a tendency to prevent the increase, indicating that lower doses of OP 2507 may promote a reaction catalyzed by xanthine oxidase. We propose that both the co-administration of prostaglandins and other potentially hepatotoxic drugs, and the administration of prostaglandins to patients with drug-induced liver damage should be done carefully. PMID- 8727711 TI - Comparative study of C-reactive protein in chronic hepatitis B and chronic hepatitis C. AB - C-reactive protein (CRP) is a liver-specific acute-phase protein, and its expression in hepatocyte is regulated by cytokines such as interleukin-1, interleukin-6 and tumor necrosis factor-alpha. Although several alterations in cytokines have been found in patients with chronic viral hepatitis, it remains obscure how CRP expression is associated with progression of the disease in chronic viral infection. In the present study, CRP expression was evaluated in 45 patients with chronic hepatitis B and in 38 patients with chronic hepatitis C. By the immunohistochemical analysis, the intensity of CRP expression in hepatocyte was closely associated with the histology activity index (HAI) score in chronic hepatitis B. In contrast, the association was not found in chronic hepatitis C. When serial changes in serum levels of CRP were compared in long-term follow-up patients including 5 patients with chronic hepatitis B and 4 patients with chronic hepatitis C, serum levels of CRP fluctuated simultaneously with serum levels of alanine aminotransferase in chronic hepatitis B, whereas the correlation was not recognized in chronic hepatitis C. These results suggest that CRP expression correlates with progression of the disease in chronic hepatitis B, but not in chronic hepatitis C. It is also possible that cytokine-mediated response is more pronounced in chronic hepatitis B than in chronic hepatitis C. PMID- 8727712 TI - Resident gynecologists and total hysterectomy. AB - We examined the specific number of surgeries necessary for a three-year obstetrics and gynecology resident to acquire proficiency in two types of hysterectomies. Improvement in the technical skills of the residents was assessed using surgical time and blood loss, and resected tumor weight was chosen as the factor representing the difficulty of the surgery. Regarding abdominal total hysterectomy (ATH), early residents (less than 25 ATH experience) performed relatively easier surgeries, and improvement in technical skill was manifested as reduced blood loss by mid residents (25 to 49 ATH) and as shortened surgical time by later residents (75 or more ATH). Regarding vaginal total hysterectomy (VTH), blood loss for earlier residents (less than 15 VTH) was greater than that for the staff, and there was a significant difference between staff surgical time and that for each resident group. These data suggest that performing more than 75 ATH during the residency period of three years is adequate to establish proficiency in this type of surgery, but that the execution of 25 VTH is insufficient and that residents require more training to learn VTH. PMID- 8727713 TI - Secondary carnitine palmitoyltransferase deficiency in chronic renal failure and secondary hyperparathyroidism. AB - A 14-year-old girl, having mental and growth retardation with end stage renal disease, was affected by a stroke-like attack. The attack was associated with transient low density areas at both sides of the parietal portion on head CT. Lactic acidosis, hypertrophic cardiomyopathy, angina pectoris-like attacks, hypertension and hyperparathyroidism were also observed and they were supposedly due to mitochondrial cytopathy. No morphological or biochemical abnormalities were found on the mitochondrial respiratory chain. However, muscle carnitine palmitoyltransferase (CPT) activity was significantly low, which was restored to a normal level after hyperparathyroidism was controlled by alphacalcidol administration. Furthermore, we also found two more chronic renal failure patients with secondary hyperparathyroidism, as well as the primary hyperparathyroidism patient showing markedly low muscle CPT activity. These findings suggest the possible contribution of parathyroid hormone to lipid metabolism in skeletal muscle and to the myopathic manifestations often seen in hyperparathyroidism. PMID- 8727714 TI - In situ hybridization of stromelysin mRNA in the synovial biopsies from rheumatoid arthritis. AB - We examined the expression of stromelysin mRNA (SL mRNA) in synovial biopsy specimens from 12 cases of rheumatoid arthritis (RA) and 2 cases of osteoarthritis (OA) using in situ hybridization. The study demonstrated that positive cells with high levels of SL mRNA were mostly (85%) found in the synovial lining layer. The positive cells were abundant in the synovium of RA which presented well developed lymphoid follicles with massive inflammatory cells. On the other hand, the synovium of OA contained no positive cells for SL mRNA. In addition, low yet positive levels of SL mRNA were detected in the endothelial cells and vascular myocytes, and interstitial cells in the deeper layer of the synovium. Karyometric studies showed that cells positive for SL mRNA had significantly larger and more spherical nuclei than weakly positive or negative cells. The SL mRNA positive cells did not demonstrate any immunoreactivity to markers of bone marrow origin, such as Leu M1, Leukocyte Common Antigen (LCA) and lysozyme antigen. Electron microscopy of a case with many SL mRNA positive cells showed that most had well developed rough endoplasmic reticulum and numerous processes on the cell surface, and some had also well developed rough endoplasmic reticulum but without processes indicating that they may be AB and/or B synoviocytes. PMID- 8727715 TI - Recurrent breast cancer treated successfully with mitomycin-C and vinblastine after failure of both doxorubicin-containing regimen and paclitaxel--a case report. AB - Cross-resistance is one of the chief obstacles in salvage therapy for refractory breast cancer. Although paclitaxel is one of the most promising drugs, it shows a response rate of 30% at most for patients with breast cancer resistant to doxorubicin, and no effective treatments for tumors refractory to both agents have been reported. We describe a 38-year-old woman with recurrent breast cancer, who was treated successfully with mitomycin-C and vinblastine after doxorubicin based chemotherapy and paclitaxel failed. The combinations of mitomycin-C and microtubule inhibitors including vinca alkaloids and taxanes may have a potential application to refractory breast cancer. PMID- 8727716 TI - Juvenile adenomyotic cyst of the corpus uteri with dysmenorrhea. AB - The clinical and pathological features of an apparently unique case of an endometrial cyst of the uterus are reported. The cyst was located within the myometrium of a 16-year-old woman suffering from dysmenorrhea. After excision of the cyst, patient's symptoms improved. On histological examination, the cyst most closely resembled an adenomyotic cyst. PMID- 8727717 TI - Carpal instability. AB - A great deal of progress has been made in recent years with respect to understanding the normal and pathologic anatomy of the wrist. Nonetheless, our knowledge is incomplete, so there still is room for diversity of opinion regarding the diagnosis and treatment of most of the presently recognized wrist instabilities. A careful history and physical examination combined with a critical review of the standard radiographs, supplemented by additional studies as indicated, allow the astute clinician to identify specific patterns of instability and to formulate an effective treatment program for the patient. PMID- 8727718 TI - Evaluation and treatment of the rheumatoid wrist. PMID- 8727719 TI - Acute dislocations of the distal radioulnar joint. AB - An injury to the DRUJ can occur in association with almost any fracture of the forearm or as an isolated phenomenon. A dislocation of this joint may be simple or complex. Constant awareness must be maintained during treatment of injuries of the wrist, forearm, and elbow if common pitfalls are to be avoided. Failure to diagnose and treat a complex DRUJ dislocation will lead to chronic, persistent subluxations or dislocations, or both, and to symptomatic osteoarthrosis. Failure to recognize a simple dislocation of the DRUJ associated with a fracture of the forearm may result in inappropriate or inadequate immobilization of the dislocated joint after internal fixation of the fracture. As a consequence, the injured TFCC may not heal, leading to recurrent post-operative instability. After either a simple or a complex dislocation of the DRUJ has been recognized, the treatment is relatively straightforward and rewarding. Despite the severity of these injuries, with proper diagnosis and management most patients will have a satisfactory outcome. PMID- 8727720 TI - Hand infections in patients with diabetes. PMID- 8727721 TI - The arthroscopic treatment of anterior and multidirectional shoulder instability. PMID- 8727722 TI - Arthroscopic transglenoid suture capsulorrhaphy for anterior shoulder instability. AB - The role of arthroscopic procedures in the management of glenohumeral stability continues to evolve. Arthroscopic transglenoid suture capsulorrhaphy, as described, is an effective alternative for correcting the pathology of anterior and anteroinferior shoulder instability. The procedure avoids the morbidity associated with open repairs as a result of anterior soft tissue dissection and division of the subscapularis. The procedure, however, is technically demanding. To achieve predictable and reproducible results that approach those seen in comparable patient populations undergoing open Bankart stabilization, recognition of, and addressing, the responsible pathology is essential, as is appropriate patient selection. Based upon experience to date, arthroscopic transglenoid suture capsulorraphy is best indicated for traumatic anterior dislocators with a Bankart lesion and good quality glenohumeral ligaments. Arthroscopic suture capsulorrhaphy is not recommended for patients involved in upper extremity collision sports, those with a large glenoid rim defect, habitual luxators, patients younger than 18 years of age, and for surgeons whose experience with operative arthroscopy of the shoulder is limited. The procedure has an unresolved role, due to lack of sufficient long-term data in acute dislocations, multidirectional instability, those with previously failed open or arthroscopic repairs, and those with generalized joint hyperlaxity. The eventual role of arthroscopic transglenoid suture capsulorrhaphy in the treatment of anterior shoulder instability awaits further studies and technological innovation. PMID- 8727723 TI - Arthroscopic evaluation and treatment of injuries to the superior glenoid labrum. PMID- 8727724 TI - Arthroscopic treatment of anterior shoulder instability using threaded suture anchors and nonabsorbable suture. PMID- 8727725 TI - Arthroscopic evaluation of acute initial shoulder dislocations. PMID- 8727726 TI - Arthroscopic shoulder stabilization using a bioabsorbable fixation device. PMID- 8727727 TI - Anterior and posterior spinal surgery: same-day, staged, anterior first, posterior first, or simultaneous? PMID- 8727728 TI - Where to stop the fusion distally in adult scoliosis: L4, L5, or the sacrum? AB - Multiple considerations must be weighed and discussed with the patient before deciding on a long fusion down to the middle or distal lumbar spine. The factors outlined above all play a role in the outcome of such surgery. PMID- 8727729 TI - Load sharing principles: the role and use of anterior structural support in adult deformity. AB - The goals of load sharing are to restore balance between posterior tension and anterior compression, ideally to maintain the instantaneous axis of rotation just posterior to the anulus and to thereby minimize the forces on the reconstruction. To accomplish this, the surgeon should understand the anatomic column deficiencies of the patient's pathology and the appropriate options for anterior and posterior reconstruction. PMID- 8727730 TI - Congenital spinal deformity. PMID- 8727731 TI - Freeze-dried cortical strut allografts for femoral reconstruction in revision hip replacement surgery. PMID- 8727732 TI - Acetabular allografts for restoration of bone stock in revision arthroplasty of the hip. PMID- 8727733 TI - Proximal femoral allografts for reconstruction of bone stock in revision arthroplasty of the hip. PMID- 8727734 TI - Acetabular reconstruction with massive acetabular allografts. PMID- 8727735 TI - Metal ring supports for deficient acetabular bone in total hip replacement. PMID- 8727737 TI - Bone cement as a seal protecting the femur from the ingress of particulate debris and femoral osteolysis. PMID- 8727736 TI - Periprosthetic osteolysis in total hip arthroplasty: the role of particulate wear debris. PMID- 8727738 TI - The role of polyethylene properties in osteolysis after total hip replacement. PMID- 8727739 TI - Classification of bone loss in total hip arthroplasty. PMID- 8727740 TI - Osteotomies about the hip for the prevention and treatment of osteoarthrosis. PMID- 8727741 TI - Biology of ACL reconstructions: what happens to the graft? PMID- 8727742 TI - Anterior cruciate ligament: injuries and treatment. PMID- 8727743 TI - ACL reconstruction using the autogenous bone-patellar tendon-bone graft: open two incision technique. PMID- 8727744 TI - Prevention of complications after autogenous bone-patellar tendon-bone ACL reconstruction. PMID- 8727745 TI - Rehabilitation after autogenous bone-patellar tendon-bone ACL reconstruction. AB - In the past decade, changes in the rehabilitation of patients with ACL injuries have made a dramatic impact on the management of the surgically treated ACL deficient knee. Our rehabilitation program following the ACL reconstruction has been developed over the past 12 years. With close follow-up of our patients, we have modified the rehabilitation program in an attempt to eliminate postoperative complications, ensure long-term knee stability, and allow a safe and rapid return to athletic activities. Our present rehabilitation protocol is divided into four phases. The initial phase encompasses the preoperative period. The second phase involves the first 2 weeks after ACL reconstruction. The third phase extends from 2 to 5 weeks postoperation, and the final phase (> than 5 weeks postoperation) involves a safe return to athletic activities. An aggressive preoperative phase and close attention to full hyperextension, control of swelling, and good leg control during the first postoperative phase are the cornerstones of the rehabilitation program. We continue to evaluate our results closely and to advance our rehabilitation program guided by our patients and continuous follow up evaluations. We propose that the avoidance of immediate surgery and appropriate preoperative patient preparation, along with postoperative rehabilitation program that emphasizes extension and closed chain function exercises, will optimize the final result. PMID- 8727746 TI - Management of combined injuries of the anterior cruciate and medial collateral ligaments. PMID- 8727747 TI - Outpatient ACL surgery: a review of safety, practicality, and economy. PMID- 8727748 TI - Arthroscopically assisted ACL reconstruction using central third patellar tendon autograft with press fit femoral fixation. AB - The ACL is a complex structure, and its replacement involves precise surgical technique, which relates directly to its function in constraining anterior tibial translation. Many techniques have been developed for reconstruction of the ACL. Here we have described our method of choice for arthroscopically assisted graft placement, which uses the middle-third patellar tendon autograft placed using a press-fit fixation technique. This technique was developed to avoid drawbacks encountered with fixation using an interference screw. Biomechanical testing has shown this technique to be similar in pull-out strength to fixation obtained with an interference screw, but without the associated technical difficulties. One other benefit of this technique, which is not readily apparent, and not one that we, as surgeons, would ever hope to take advantage of, is that should a revision ACL reconstruction be necessary in the future, there is no interference screw in the femur to remove. PMID- 8727749 TI - Intraoperative complications of arthroscopically assisted ACL reconstruction using patellar tendon autograft. PMID- 8727750 TI - Factors influencing the longevity of UHMWPE tibial components. PMID- 8727751 TI - Congenital vertical talus. PMID- 8727752 TI - Tarsal coalitions. PMID- 8727753 TI - Controversies in the surgical management of clubfoot. PMID- 8727754 TI - Clubfoot controversies: complications and causes for failure. PMID- 8727755 TI - Flexible flatfoot and skewfoot. PMID- 8727756 TI - Bunions and deformities of the toes in children and adolescents. PMID- 8727757 TI - Bone biology. I: Structure, blood supply, cells, matrix, and mineralization. PMID- 8727758 TI - Bone biology. II: Formation, form, modeling, remodeling, and regulation of cell function. PMID- 8727759 TI - Enhancement of fracture healing. PMID- 8727760 TI - Chondrosarcoma: a review. PMID- 8727761 TI - Benign bone tumors. PMID- 8727762 TI - Evaluation, diagnosis, and classification of benign soft-tissue tumors. PMID- 8727763 TI - Soft-tissue sarcomas. PMID- 8727764 TI - Cerebral palsy: orthopaedic management. PMID- 8727765 TI - Gait analysis: principle and applications with emphasis on its use in cerebral palsy. PMID- 8727766 TI - Esoteric contact dermatitis. Part III: Ragweed dermatitis. PMID- 8727767 TI - Ultraviolet light and immunity to coelenterate venom. AB - Peripheral blood lymphocytes from sensitized persons proliferate in response to naturally occurring (coelenterate) venoms. Ultraviolet exposure decreases the proliferation. This decrease is widespread and also extends to responses against pathogens, mitogens, and endogenous proteins. Ultraviolet exposure alters human immunity by many means, including affecting circulating T-cells. Since venoms act by immunologic as well as toxic pathways, any abrogation of normal immunity could enhance their action on man. PMID- 8727768 TI - LEOPARD syndrome (cardiocutaneous lentiginosis syndrome). PMID- 8727769 TI - Familial Kaposi's sarcoma. AB - Familial cases of Kaposi's sarcoma are exceedingly rare. We identified an 85-year old man with Kaposi's sarcoma. Multiple lesions were present on the feet. Previously, the man's 84-year-old sister presented with similar lesions on the lower extremity. Both siblings were lifelong residents of the United States and were of English/German descent. Neither had risk factors for infection with human immunodeficiency virus (HIV). On histologic examination, lesions in both patients demonstrated vascular slits, hemorrhage, and dense aggregates of spindle cells within the dermis. PMID- 8727770 TI - Dilantin hypersensitivity reaction. AB - Dilantin hypersensitivity syndrome is characterized by fever, rash, lymphadenopathy, facial edema, and hepatitis. Anemia, pharyngitis, diarrhea, and nephritis may also be present. The eruption may present as the classic erythematous follicular papules and pustules; or it may be pleomorphic, presenting as a morbilliform eruption, erythroderma, or toxic epidermal necrolysis. Early recognition and discontinuation of the medication are necessary to prevent a potentially fatal outcome. A case is presented and the literature reviewed. PMID- 8727771 TI - Cutaneous cryptococcosis in a patient receiving chronic immunosuppressive therapy. AB - Cryptococcosis was diagnosed in an immunosuppressed patient. Topical treatment was ineffective and treatment with oral itraconazole was started. The lesion improved. Liver enzymes were monitored and, although their levels were raised before the start of itraconazole therapy, they did not increase further during treatment. PMID- 8727773 TI - Contact allergy testing alternatives: 1996. AB - The recent release of an easy-to-apply patch test system has made standard patch testing more practical for the busy office practitioner. This is an important advance since contact dermatitis is being increasingly recognized as a major cause of eczema recalcitrant to treatment. The identification of responsible allergens is, however, only the first step in helping the patient with contact allergy. This review lists appropriate alternative materials that can safely be used on persons allergic to each material on the standard tray of allergens. PMID- 8727772 TI - Acyclovir in the treatment of hand-foot-and-mouth disease. AB - Twelve children ages 1 to 5 years and one adult with hand-foot-and-mouth disease were treated with oral acyclovir within one to two days of onset of the rash. Symptomatic relief, defervescence, and significant involution of lesions were seen within twenty-four hours of initiating therapy. Acyclovir was continued for five days, by which time palmar, plantar, and oral lesions were virtually gone. Acyclovir is a molecule tailored to inactivate the thymidine kinase of the herpesvirus. Since the Coxsackie A16 virus causing hand-foot-and-mouth disease lacks this enzyme, the beneficial therapeutic effect must be explained on other grounds, possibly due to enhancement of the antiviral effect of the patient's own interferon. PMID- 8727774 TI - Knuckle pads--a forgotten skin condition: report of a case and review of the literature. AB - A 12-year-old boy presented with knuckle pads, palmar keratoderma, and hand eczema. Since Morginson's description of knuckle pads associated with other dermatoses almost forty years ago, little attention has been given to this disorder. We describe a case and review some of the clinical aspects of this disorder. PMID- 8727775 TI - Blue nails and acquired immunodeficiency syndrome: not always associated with azidothymidine use. AB - Nail dyschromia in patients infected with human immunodeficiency virus (HIV) was first described in 1987 by Furth and Kazakis. It has since been reported in many patients with the acquired immunodeficiency syndrome (AIDS), predominantly in those taking azidothymidine. There have been only three reports of nail pigmentation in HIV-infected patients who had not received azidothymidine. We describe such a case. PMID- 8727776 TI - Stump pemphigoid. AB - Stump pemphigoid is a localized variant of bullous pemphigoid characterized by specific clinical, pathological, and immunofluorescent characteristics. The early diagnosis of this entity guides treatment toward this specific immunologically mediated inflammatory process and away from treatment of unrelated conditions with similar clinical findings. PMID- 8727778 TI - Sexually induced dermatoses. AB - Sexually induced dermatoses (SID) are conditions resulting from sexual activity. Sometimes this can be modified by bacterial infection or systemic disease. Many materials such as medications, cosmetics, perfumes, pads, hygiene, sprays, condoms, etc, may cause sexually induced dermatoses in sexual partners. Sexually induced dermatoses are noninfectious, self-limited conditions. PMID- 8727777 TI - Diffuse histiocytic lymphoma treated with chemotherapy and radiation. AB - A case of cutaneous lymphoma treated aggressively with chemotherapy and radiation is presented. This case is particularly interesting due to the fact that the patient also had a history of a malignant melanoma removed from the right shoulder, 1.26 mm thick, invasive to level III. He had two subsequent right axillary node tumors and a mesenteric tumor removed, all revealing metastatic malignant melanoma. PMID- 8727779 TI - Piezogenic pedal papules in two family members. AB - Piezogenic pedal papules are small, occasionally painful, fat herniations that become apparent when weight is placed on the heel. Although the cause is unknown, hereditary factors may play a role. This is the second report of two family members with this condition. PMID- 8727780 TI - Skin metastases from a laryngeal carcinoma: report of a case. AB - We report a patient who showed skin metastases a few months after the diagnosis and treatment of a laryngeal carcinoma. Review of the literature confirms its rarity and the severity of its prognosis. PMID- 8727782 TI - Bilateral inflammatory linear verrucous epidermal nevus associated with psoriasis. AB - Inflammatory linear verrucous epidermal nevus is a benign hyperplasia of the epidermis. Bilateral distribution of inflammatory linear verrucous epidermal nevus is an extremely rare presentation. Although inflammatory linear verrucous epidermal nevus is generally accepted as a distinct entity, it has similar clinical and histopathologic features to psoriasis, or they occasionally may overlap. We report the case of an 8-year-old boy with bilateral inflammatory linear verrucous epidermal nevus who also had psoriasis. PMID- 8727781 TI - Cutis laxa acquisita associated with multiple myeloma: a case report and review of the literature. AB - Cutis laxa acquisita is a rare disorder that affects collagen and elastin metabolism. The cause is unknown. Characteristic features include sagging and laxity of the skin, as well as involvement of the lungs, heart, gastrointestinal system, and urogenital tract. Three cases of cutis laxa acquisita have been reported in association with multiple myeloma. Due to the rarity of these disorders, a linkage has been postulated. The clinical and histologic data from the fourth case of cutis laxa acquisita associated with multiple myeloma were compared to the three other cases previously reported in the literature. The relationship between acquired cutis laxa and multiple myeloma is unclear, with only one case revealing possible immune-mediated elastin destruction via IgG immunoglobulin bound to dermal elastin fibers on immunofluoresence examination. No pattern in the clinical courses of the disorder can be seen on review of the four cases with coincident disease. We hypothesize that cutis laxa acquisita represents a paraneoplastic process of multiple myeloma, given the rarity of these diseases. Further investigation is necessary to determine the underlying linkage between these disorders. We suggest that serum and urine protein electrophoresis results be obtained in patients presenting with cutis laxa acquisita to screen for multiple myeloma given this association. PMID- 8727783 TI - Identification and characterization of a divalent cation-dependent glycosaminoglycan-binding protein from rat liver endothelium. AB - Rat liver endothelial cells (LECs) express a membrane-associated Ca2+-dependent hyaluronan-binding activity (CaHA-BP) which is distinct from the Ca2+ independent, endocytic LEC HA receptor (Yannariello-Brown et al., J. Cell Biochem., 48, 73-80, 1992). The CaHA-BP is specific for a subset of glycosaminoglycans, since Ca2+-dependent binding of 125I-HA (approximately 80 kDa) to LECs was competed with a 100-fold excess (w/w) of HA, chondroitin sulfate, and heparin, but not with chondroitin. The CaHA-BP activity on intact LECs was pH-dependent. Optimal binding occurred at pH 6.0; no binding was detected at pH values or= 9. 125I-HA, pre-bound in the presence of Ca2+, could also be dissociated with an acidic buffer (pH 5.0), as well as the divalent cation chelators EDTA and EGTA. 125I-HA binding was stimulated by divalent cations other than Ca2+, such as Mg2+, Mn2+, and Ba2+; with the exception of Zn2+. A photoaffinity crosslinking reagent (125I-ASD-HA) was used to identify specifically crosslinked polypeptides on LECs. In the absence of Ca2+, and in the presence of EGTA, only bands at 175/166 kDa were consistently crosslinked. These bands have been previously identified as the LEC Ca2+ independent endocytic HA receptor (Yannariello-Brown et al., J. Biol. Chem., 267, 20451-20455, 1992). In the presence of Ca2+, crosslinking was consistently seen to a 68 kDa polypeptide. Crosslinking was competed with a 100-fold excess (w/w) of HA. These and other data suggest that a 68 kDa protein is the most likely candidate for the CaHA-BP in LECs. PMID- 8727784 TI - Heparan sulphate/heparin glycosaminoglycans with strong affinity for the growth promoter spermine have high antiproliferative activity. AB - Depletion of intracellular polyamine pools inhibits cell proliferation. Polyamine pools are maintained by intracellular synthesis and by uptake from the extracellular environment. It may be expected that cationic polyamines are sequestered by the polyanionic glycosaminoglycan substituents of extracellular proteoglycans. Moreover, highly sulphated heparin-related glycans inhibit growth of human embryonic lung fibroblasts. We have therefore investigated interactions between polyamines and heparin-related glycosaminoglycans. Affinity chromatography of various polyamines on heparin-agarose indicated that spermine was the only polyamine that bound efficiently to this type of glycan. By using equilibrium dialysis we found that spermine binds to a highly sulphated heparan sulphate/heparin preparation with a dissociation constant of 3.7 x 10(-5)M. Enzymatic degradation of heparan sulphate using three different heparan sulphate/heparin lyases, separately or in combination and in the absence or presence of spermine, was used to generate spermine-binding and degradation protected oligosaccharides. As indicated by chromatographic and electrophoretic analysis a size- and chargeheterogeneous collection was obtained. However, protected oligosaccharides derived from antiproliferative heparan sulphates were inactive. Highly sulphated, antiproliferative heparan sulphates were subfractionated on spermine-agarose yielding high-affinity material with increased antiproliferative activity. A very potent material was obtained from pig skin. Although there was generally a clear correlation between high spermine affinity and strong growth-inhibition, no correlation with sulphate content or oligosaccharide mapping patterns could be detected. Beef lung heparan sulphate comprised naturally occurring fragments of eicosasaccharide size with substantially increased specific activity. As these fragments were longer than oligosaccharides generated by enzymatic degradation in the presence of spermine (hexa- to tetradecasaccharide), multiple spermine-binding sites in tandem may be necessary to induce antiproliferative activity. PMID- 8727785 TI - Characterization of carbohydrate structural features recognized by anti arabinogalactan-protein monoclonal antibodies. AB - Arabinogalactan-proteins (AGPs) are a diverse class of plant cell surface proteoglycans implicated in a range of fundamental processes associated with plant cell development. Anti-AGP monoclonal antibodies have been used extensively for the investigation of the developmental regulation of AGPs although virtually nothing is known about the structure of the carbohydrate epitopes recognised by these antibodies. In this report, a series of methyl glycosides of monosaccharides and a range of oligosaccharides that are elements of the carbohydrate component of AGPs have been investigated for recognition by previously derived anti-AGP monoclonal antibodies. No clear evidence was obtained for the involvement of terminal arabinofuranosides, nor of the galactan backbone, in the recognition of the glycan structure of AGPs by any of the antibodies used in this study. Interestingly, the most effective inhibitor of the binding of the monoclonal antibodies MAC207, JIM4 and JIM13 to exudate gum antigens was an acidic trisaccharide, isolated from a partial acid hydrolysate of gum karaya which has the structure: GlcA beta(1-->3) GalA alpha(1-->2)Rha, determined by a combination of FAB-MS, GC-MS and NMR spectroscopy. PMID- 8727786 TI - Tracheo-bronchial mucin gene expression as detected by in situ hybridization. AB - To understand regulation of the tracheo-bronchial mucin (TBM) gene expression we developed cDNA probes encoding TBM. We also raised antisera against mucin protein (deglycosylated and glycosylated), and developed immortalized tracheal epithelial cells which express mucin (at the RNA and protein level). TBM cDNA probes can detect TBM mRNA in situ in samples from dog and human primary tracheal epithelial cells and cell lines derived from them. For clinical application, conditions were optimized for detection of the TBM mRNA in human turbinal and nasal polyps and trachea of cystic fibrosis (CF) and non-CF subjects. Fixing and hybridization conditions were found to be critical for the optimum hybridization signal. Riboprobes proved to be better than cDNA or oligonucleotide probes. The application of these newly developed molecular tools in the genetic therapy of CF is discussed herein. PMID- 8727787 TI - Isolation and structural characterization of fucosylated gangliosides with linear poly-N-acetyllactosaminyl chains from human granulocytes. AB - The isolation and structural characterization of fucosylated neolacto-series gangliosides with linear poly-N-acetyllactosaminyl chains from normal human granulocytes is described. Gangliosides were purified by consecutive use of anion exchange HPLC on Fractogel TMAE-650(S), adsorption and reversed phase HPLC on Nucleosil 50-7 and Nucleosil 7C18 columns, respectively. TLC immunostaining with carbohydrate specific monoclonal antibodies, fast atom bombardment-mass spectrometry (FAB-MS) of the permethylated derivatives and gas chromatography electron impact mass spectrometry (GC-EIMS) of partially methylated alditol acetates were used for structure elucidations. One ganglioside was identified as sialyl Lewis(x) antigen with nLcOse6Cer core, Neu5-Ac alpha 2-3Gal beta 1-4 (Fuc alpha 1-3)GlcNAc beta 1-3Gal beta 1-4Glc NAc beta 1-3 Gal beta 1-4Glc beta 1 1Cer. Furthermore, monosialylated ceramide deca-, undeca-, dodeca- and tridecasaccharides with three (nLcOse8Cer) and four (nLcOse10Cer) linear lactosaminyl repeats were identified, carring one to three fucoses. The ceramide portions were found to contain C18 sphingosine and predominantly C16:0 fatty acids. All monosialogangliosides were homogenous concerning their terminal alpha 2-3 Neu5Ac-sialylation, but different in their fucosylation status. Beside VI3Neu5Ac, V3Fuc-nLcOse6Cer, in two of the fucosylated polylactosaminyl ganglioside fractions the sialyl Lewis(x) epitope was found, whereas five species expressed the terminal VIM-2 motif. The role of protein linked sialy Lewis(x) epitope of human granulocytes as a ligand for endothelial leukocyte adhesion molecule-1 (ELAM-1; E-selectin) and platelet activation-dependent granule external membrane protein (PADGEM; P-selectin) is well documented. However, the involvement of endothelial cells E-and/or P-selectin mediated cell-cell adhesion via lipid bound sialyl Lewis(x) and/or VIM-2 epitopes on human granulocytes has to be proved in further investigations. PMID- 8727788 TI - Glycosylation in lepidopteran insect cells: identification of a beta 1-->4-N acetylgalactosaminyltransferase involved in the synthesis of complex-type oligosaccharide chains. AB - The choice for a heterologous expression system to produce glycoprotein therapeutics highly depends on its potential to perform mammalian-like posttranslational modifications such as glycosylation. To gain more insight into the glycosylation potential of the baculovirus mediated insect cell expression system, we have studied the expression of glycosyltransferases involved in complex-type N-glycosylation. Lepidopteran insect cell lines derived from Trichoplusia ni, Spodoptera frugiperda, and Mamestra brassicae were found to express a beta 1-->4- N-acetylgalactosaminyltransferase (beta 4-GalNAcT) that catalyzes the transfer of GalNAc from UDP-GalNAc to oligosaccharides and glycoproteins carrying a terminal beta-linked GlcNAc residue. These results suggest that Lepidopteran insect cells are capable of synthesizing complex-type carbohydrate chains containing GalNAc beta 1-->4GlcNAc (LacdiNAc) units. Baculovirus infection of the cells, however, resulted in a decrease in the activity of beta 4-GalNAcT from 80 to <1 pmol.min-1 mg-1 protein within 48 h post infection. Furthermore, considerable beta-N-acetylgalactosaminidase and beta-N acetylglucosaminidase activity was observed in insect cells, whether or not infected with baculovirus, as well as in the culture medium. These enzyme activities could be responsible for degradation of complex-type oligosaccharide chains containing LacdiNAc units. Our findings provide an enzymatic basis for the observation that most recombinant glycoproteins produced by baculovirus infected insect cells carry oligomannosidic-type N-linked glycans, in spite of the fact that uninfected insect cells have the potential for the synthesis of mammalian like complex-type glycans. PMID- 8727789 TI - Elongation of the N-glycans of fowl plague virus hemagglutinin expressed in Spodoptera frugiperda (Sf9) cells by coexpression of human beta 1,2-N acetylglucosaminyltransferase I. AB - Spodoptera frugiperda (Sf9)-cells differ markedly in their protein glycosylation capacities from vertebrate cells in that they are not able to generate complex type oligosaccharide side chains. In order to improve the oligosaccharide processing properties of these cells we have used baculovirus vectors for expression of human (beta 1,2-N-acetylglucosaminyltransferase I (hGNT-I), the enzyme catalysing the crucial step in the pathway leading to complex type N glycans in vertebrate cells. One vector (Bac/GNT) was designed to express unmodified GNT-I protein, the second vector (Bac/tagGNT) to express GNT-I protein with a tag epitope fused to its N-terminus. In Sf9-cells infected with Bac/tagGNT virus a protein of about 50 kDa representing hGNT-I was detected with an antiserum directed against the tag epitope. HGNT-I activity was increased at least threefold in lysates of infected cells when N-acetylglucosamine (GlcNAc) free ovalbumine was used as substrate. To monitor hGNT-I activity in intact Sf9 cells, the glycosylation of coexpressed fowl plague virus hemagglutinin (HA) was investigated employing a galactosylation assay and chromatographic analysis of isolated HA N-glycans. Coexpression of hGNT-I resulted in an at least fourfold increase of HA carrying terminal GlcNAc-residues. The only structure detectable in this fraction was GlcNAcMan3GlcNAc2. These results show that hGNT-I is functionally active in Sf9-cells and that the N-glycans of proteins expressed in the baculovirus/insect cell system are elongated by coexpression of glycosyltransferases of vertebrate origin. Complete complex type oligosaccharide side chains were not observed when hGNT-I was overexpressed, thus supporting the concept that Sf9-cells do not contain glycosyltransferases acting after hGNT-I. PMID- 8727790 TI - A new approach to mapping co-localization of multiple glycosyl transferases in functional Golgi preparations. AB - We have developed a new method to co-localize multiple glycosyl transferases in different Golgi compartments. The approach relies on the proven ability of intact, sealed rat liver Golgi preparations to concentrate exogenous labeled sugar nucleotides into the lumen where they glycosylate either endogenous or artificial acceptors. The premise is that if two glycosyl transferases are co localized within the same compartment, they will compete for the limited amount of transported donor. If the donor is consumed in glycosylating a permeable artificial glycoside within a Golgi compartment, it will be unavailable to glycosylate endogenous products within that same compartment. The greater the degree of transferase co-localization, the greater the potential decrease in glycosylation of endogenous acceptors. We provide an example consistent with these predictions. Adding 1 microM UDP[3H]Gal to Golgi preparations followed by a chase with a cocktail of unlabeled sugar nucleotides labels mostly endogenous N linked oligosaccharides containing both beta 1,3- and beta 1,4[3H]Gal residues with and without sialic acid. Addition of increasing amounts of 4 methylumbelliferyl-beta-xyloside (Xyl beta MU) produces [3H]Gal1 beta, 4Xyl beta MU and leads to a reciprocal decrease in labeling of a restricted set of the endogenous acceptors. This decrease is preferential for [3H]Gal beta 1-->3GlcNAc beta 1-->R and, to a lesser extent, [3H]Gal beta 1-->4GlcNAc beta 1-->R structures in neutral and mono-sialylated oligosaccharides; synthesis of these structures in di- and tri-sialylated oligosaccharides was unaffected. These preferential decreases are not seen in detergent permeabilized, sugar nucleotide transport-independent Golgi incubations, and are not due to inhibition by the Gal beta 1,4Xyl beta MU product. These results argue that there is significant overlap in the functional co-localization of sialyl and galactosyltransferases in rat liver Golgi preparations and that GAG chain core specific Galactosyltransferase I is co-localized with subsets of N-glycan Gal beta 1,3 and Gal beta 1,4 transferases. This approach can be used with other glycosides and sugar nucleotides to map and co-localize other glycosyl transferases. The functional compartments defined by this approach may or may not correspond entirely with morphologically defined Golgi domains. PMID- 8727791 TI - Subsets of sialylated, sulfated mucins of diverse origins are recognized by L selectin. Lack of evidence for unique oligosaccharide sequences mediating binding. AB - Previous studies have shown that the mucin-type polypeptides GlyCAM-1, CD34, and MAdCAM-1 can function as ligands for L-selectin only when they are synthesized by the specialized high-endothelial venules (HEV) of lymph modes. Since sialylation, sulfation, and possibly fucosylation are required for generating recognition, we reasoned that other mucins known to have such components might also bind L selectin. We show here that soluble mucins secreted by human colon carcinoma cells, as well as those derived from human bronchial mucus can bind to human L selectin in a calcium-dependent manner. As with Gly-CAM-1 synthesized by lymph node HEV, alpha 2-3 linked sialic acids and sulfation seem to play a critical role in generating this L-selectin binding. In each case, only a subset of the mucin molecules is recognized by L-selectin. Binding is not destroyed by boiling, suggesting that recognition may be based primarily upon carbohydrate structures. Despite this, O-linked oligosaccharide chains released from these ligands by beta elimination do not show any detectable binding to L-selectin. Following protease treatment of the ligands, binding persists in a subset of the resulting fragments, indicating that specific recognition is determined by certain regions of the original mucins. However, O-linked oligosaccharides released from the subset of non-binding mucin fragments do not show very different size and charge profiles compared to those that do bind. Furthermore, studies with polylactosamine-degrading endoglycosidases suggest that the core structures involved in generating binding can vary among the different ligands. Taken together, these data indicate that a single unique oligosaccharide structure may not be responsible for high-affinity binding. Rather, diverse mucins with sialylated, sulfated, fucosylated lactosamine-type O-linked oligosaccharides can generate high-affinity L-selectin ligands, but only when they present these chains in unique spacing and/or clustered combinations, presumably dictated by the polypeptide backbone. PMID- 8727792 TI - Uptake and metabolism of BuCast: a glycoprotein processing inhibitor and a potential anti-HIV drug. AB - We have previously shown (Sunkara et al., 1989; Taylor et al., 1991) that 6-o butanoyl castanospermine (BuCast) was a 30-50-fold better inhibitor of HIV syncytia formation than castanospermine (Cast). Radiolabeled Cast and BuCast were used to study the uptake and metabolism of these compounds in cultured cells and in mice. BuCast was preferentially taken up by cells compared to Cast. Approximately 30-50-fold higher radioactivity was found in cells treated with BuCast compared to cells treated with Cast during the initial 4-6 h of labeling. HPLC analysis showed that once BuCast was taken up by cells, it was rapidly converted to Cast. Mice given oral doses of BuCast had 5-10-fold higher levels of Cast in the plasma and tissues as compared to Cast treated mice. However, when the compounds were given i.v., the levels of plasma and tissue radioactivity obtained from Cast of BuCast were equivalent suggesting rapid conversion of BuCast to Cast in the blood. In mice orally treated with BuCast, HPLC analysis suggested that only Cast was found in the plasma and tissues. With multiple dosing of mice, additive results were obtained, suggesting that multiple doses may be used to obtain higher concentrations of the compound in the target cells. These data suggest that the lipophilic properties of butanoyl side chain on the Cast ring makes BuCast significantly better absorbed, and this may help to alleviate some of the gut toxicity associated with Cast treatment. PMID- 8727793 TI - The hypothetical N-glycan charge: a number that characterizes protein glycosylation. AB - The production of recombinant glycoprotein therapeutics requires characterization of glycosylation with respect to the lot-to-lot consistency. Here we introduce the ?hypothetical N-glycan charge Z' as a parameter that allows to characterize the protein glycosylation in a simple, however, efficient manner. The hypothetical N-glycan charge of a given glycoprotein is deduced from the N-glycan mapping profile obtained via HPAE-PAD. In HPAEC, N-glycans are clearly separated according to their charge, i.e., their number of sialic acid residues, providing distinct regions for neutral structures as well as for the mono- di-, tri, and tetrasialylated N-glycans (Hermentin et al., 1992a). Z is defined as the sum of the products of the respective areas (A) in the asialo, monosialo, disialo, trisialo, tetrasialo, and pentasialo region, each multiplied by the corresponding charge: [formula: see text] Thus, a glycoprotein with mostly C4-4* structures will provide Z approximately equal to 400 (e.g., rhu EPO (CHO), Z = 361), a glycoprotein carrying largely C3-3* structures will amount to Z approximately equal to 300 (e.g., bovine fetuin, Z = 290), a glycoprotein with mostly C2-2* structures will have Z approximately equal to 200 (e.g., human serum transferrin, Z = 207, or human plasma AT III, Z = 180), and a glycoprotein carrying only high mannose type or trunkated structures will provide Z approximately equal to 0 (e.g., bovine pancreas ribonuclease B, Z = 15, and hen ovomucoid, Z = 15, respectively). The determination of Z was validated in multiple repetitive experiments and proved to be highly accurate and reliable. Z may therefore be regarded as a new and characteristic parameter for protein N-glycosylation. PMID- 8727794 TI - Organization of a human UDP-GalNAc:polypeptide, N-acetylgalactosaminyltransferase gene and a related processed pseudogene. AB - We have previously characterized a cDNA that encodes a full length human UDP GalNAc:polypeptide, N-acetylgalactosaminyltransferase (GalNAc-transferase) (J.A. Meurer et al., J. Biochem., 118, 568-574, 1995). The present report describes the characterization of the corresponding human GalNAc-transferase gene and a related pseudogene. Two human genomic libraries, lambda and P1, were screened with probes derived from the human GalNAc-transferase nucleotide sequence, resulting in the isolation of four genomic clones. Southern blotting, PCR analysis, and sequencing revealed that three clones, lambda.HG-5, P1.GALN-A, and P1.GALN-B, contained overlapping genomic sequences that encompass over 55 kilobase pairs (kb) of genomic DNA and comprise a portion of the human GalNAc-transferase 5'-and 3' untranslated regions and the entire coding region. The human GalNAc-transferase gene structure consists of at least 11 exons ranging in size from 99 to > 620 nucleotides which are separated by 10 introns ranging in size from 0.7 to approximately 12.5 kb. The fourth genomic clone, P1-GALN-psi, contained a approximately 2.4 kb sequence region which shares an overall 78.6% nucleotide identity with coding region exons 1 and 3 through 11 of the human GalNAc transferase gene. However, a lack of intron sequences, as well as the presence of multiple nucleotide mutations, insertions, and deletions that disrupt the potential GalNAc-transferase reading frame, suggest that P1.GALN-psi contains a processed pseudogene. Screening of a human/rodent somatic cell hybrid panel with a P1.GALN-psi probe localized the GalNAc-transferase pseudogene to chromosome 3. Hence, the human genome contains at least two related GalNAc-transferase genes that are located on separate chromosomes. PMID- 8727795 TI - Biotherapy and the oncology nurse. AB - OBJECTIVE: To provide a brief overview of biotherapy and the roles of the oncology nurse. DATA SOURCES: Published articles that pertain to biotherapy and implications for oncology nurses. CONCLUSIONS: Biotherapy is the fourth modality of cancer therapy, and it can be effective alone or in combination with surgery, radiotherapy, and chemotherapy. Many new roles and opportunities for oncology nurses will result from the further development of biotherapy. IMPLICATIONS FOR NURSING PRACTICE: Professional nurses have an obligation to obtain accurate and current knowledge of new treatments. Oncology nurses with expertise in biotherapy will be needed to manage care for patients receiving biotherapy. PMID- 8727796 TI - The immune system. AB - OBJECTIVE: To explain the role of the immune system in cancer control and its response to environmental and perceived stressors. DATA SOURCES: Review articles, research studies, and book chapters related to immunology and the immune system. CONCLUSIONS: The immune system responds to foreign pathogens and cancer cells by activating specific and nonspecific immune responses. The goal of immunotherapy is to enhance these responses to control the growth of cancer cells. Knowledge of the influence of stress on immune and cytokine response is evolving. IMPLICATIONS FOR NURSING PRACTICE: Knowledge of the principles of immunology, the immune response to cancer, the role of cytokines in mediating immune response, and the influence of stress on immune and cytokine response will help nurses provide quality care to patients receiving biological agents. PMID- 8727797 TI - The interferons. AB - OBJECTIVE: To provide a review of the biology, pharmacology, clinical application, and toxic effects of interferon and strategies for nursing management. DATA SOURCES: Research studies, review articles, and book chapters pertaining to interferons. CONCLUSIONS: Interferons are active therapeutic agents for hairy cell leukemia, chronic myelogenous leukemia, melanoma, and acquired immunodeficiency syndrome-related Kaposi's sarcoma. Clinical trials continue to investigate the clinical application of interferons. IMPLICATIONS FOR NURSING PRACTICE: Oncology nurses can facilitate patient self-care in managing side effects of interferon therapy, provide patient and family education, and assist with reimbursement concerns. PMID- 8727798 TI - Interleukins: the search for an anticancer therapy. AB - OBJECTIVES: To review each of the interleukins, identify their common toxic effects, and highlight the potential role of interleukins in cancer therapy. DATA SOURCES: Research articles, abstracts, review articles, and book chapters related to interleukins. CONCLUSIONS: Interleukins play a key role in coordinating complex immune defense mechanisms against microbe invasion and cancer. The process of genetic engineering has made an increasing number of these interleukins available as biopharmaceuticals for evaluation of their potential roles as a cancer therapeutic agent. IMPLICATIONS FOR NURSING PRACTICE: Nurses are challenged to understand and manage better both common and unusual side effects of interleukin therapies and provide appropriate patient and family education. PMID- 8727799 TI - Hematopoietic growth factors in clinical practice. AB - OBJECTIVE: To review the hematopoietic growth factors (HGFs) as they relate to hematopoiesis, dosing, administration, toxicities and clinical indications. DATA SOURCES: Research studies, abstracts, review articles, product package inserts, and book chapters pertaining to HGFs. CONCLUSIONS: HGFs are essential components for the development of blood cells. Recombinant DNA technology and the availability of HGFs have significantly altered the care of patients with cancer and bone marrow disorders. IMPLICATIONS FOR NURSING PRACTICE: Nurses can play a major role in assisting patients, families, and various consumers by providing education about HGFs as well as their indications, schedule, methods of administration, and toxicities. Familiarity with the HGFs and the literature available can assist nurses in developing management strategies for patients receiving these agents. PMID- 8727800 TI - Monoclonal antibodies: innovations in diagnosis and therapy. AB - OBJECTIVE: To provide a review of the use of monoclonal antibodies (MAbs) for diagnosis and therapy, the obstacles affecting their usefulness, and nursing considerations unique to MAb-based treatments. DATA SOURCES: Research studies, review articles, and book chapters pertaining to MAbs in cancer diagnosis and therapy. CONCLUSIONS: Recent clinical trials have shown the potential of MAbs as diagnostic and therapeutic tools in cancer. MAbs may be used alone or conjugated with drugs, radionuclides, and toxins; however numerous obstacles limit their effectiveness. IMPLICATIONS FOR NURSING PRACTICE: Oncology nurses caring for patients receiving MAb-based therapies play an important role in the implementation of the patient's regimen. Hypersensitivity reactions, constitutional symptoms, and serum sickness are toxic effects that require monitoring and patient education. PMID- 8727801 TI - Gene therapy of cancer. AB - OBJECTIVE: To provide a review of the basic concepts and terminology of human genetics, the clinical application and risks of gene therapy, and ethical and societal issues. DATA SOURCES: Review articles, research studies, and book chapters related to genetics and human gene therapy. CONCLUSIONS: The genetic changes that play a role in the development of cancer have begun to be elucidated, and these discoveries have led to strategies to destroy cancers by correcting genetic defects or manipulating genes to induce tumoricidal activities. Gene therapy is a novel investigational intervention that is constantly evolving. IMPLICATIONS FOR NURSING PRACTICE: Oncology nurses will need an increased understanding of cellular biology, genetics, and genetics engineering. Major considerations for nursing practice include patient and family education, informed consent, side effects, and ethical and societal issues. PMID- 8727802 TI - Nursing management of patients receiving biological therapy. AB - OBJECTIVE: To provide an overview of the most common toxic effects associated with biotherapy and strategies for effective patient management, predominantly in the outpatient setting. DATA SOURCES: Review articles and book chapters related to nursing care of patients receiving biotherapy. CONCLUSIONS: Biological response modifiers can produce several toxic effects that may have a significant impact on the patient's quality of life. Additionally, biotherapy is being delivered more frequently in the ambulatory and home care settings. IMPLICATIONS FOR NURSING PRACTICE: As the role of biotherapy expands in the treatment of cancer, oncology nurses must be attuned to the special needs of patients receiving this therapy. Nurses must be knowledgeable of not only symptom management but reimbursement and managed care issues as well. PMID- 8727803 TI - Future projections in biotherapy. AB - OBJECTIVES: To review barriers to effective biotherapy and future trends in biotherapy and the expertise needed by oncology nurses to address the challenges that will result from these changes. DATA SOURCES: Review articles and book chapters related to biotherapy. CONCLUSIONS: To sustain the progress made in biotherapy, barriers that limit the effectiveness of treatment must be addressed and basic research continued. Targeted areas for new biotherapy strategies are interference with cellular communication, control of the cell cycle, and interference with the process of metastasis. IMPLICATIONS FOR NURSING PRACTICE: Nurses with creativity and motivation will continue to lead the way in developing management strategies for patients receiving future therapies. PMID- 8727804 TI - Diagnosis and treatment of superficial bladder cancer: an update. AB - It is expected that approximately 50,500 new cases of bladder cancer will be diagnosed in 1995. In addition, approximately 10,000 people are expected to die of this disease during the year. An 11% increase in incidence observed from 1957 to 1987 was accompanied by a 19.4% decrease in the death rate in American males. This rate change may be attributable to stage migration to more localized disease as well as increased survival owing to improved treatment. Transurethral resection is standard first-line treatment for transitional cell carcinoma of the bladder. Intravesical therapy with cytotoxic agents or immunomodulators has become an important element in the management of patients with superficial bladder cancer at risk for disease recurrence and progression. Study results confirm the efficacy of both mitomycin and bacille Calmette-Guerin when administered intravesically. Comparative studies of these two agents are warranted to further clarify their respective roles in the prophylaxis of recurrences of superficial bladder cancer. Studies should enroll large numbers of patients, particularly those with high-grade lesions at significant risk of recurrence and progression. Current research involves evaluation methods of optimizing the efficacy of intravesical chemotherapy. High drug concentrations, hyperthermia, and combination modalities are a few new avenues being studied. In addition, new drugs like bropirimine, interferons, mycobacteria-associated antigens, and keyhole-limpet hemocyanin are being explored. PMID- 8727805 TI - Update on the Dutch Cooperative Trial: mitomycin versus bacillus Calmette-Guerin Tice versus bacillus Calmette-Guerin RIVM in the treatment of patients with pTA pT1 papillary carcinoma and carcinoma in situ of the urinary bladder. Dutch South East Cooperative Urological Group. AB - Prophylactic intravesical treatment with chemotherapeutic agents or one of the different strains of the immunomodulator bacillus Calmette-Guerin (BCG) reduces tumor recurrence and may prevent progression after transurethral resection (TUR) of superficial bladder cancer. Clinical and pathological prognostic factors are used to categorize different groups at risk for disease recurrence and progression. Solitary low-grade (G1, 2A), low-stage (pTa) tumors have less than a 5% risk of progression, while pT1 G3 tumors with concomitant carcinoma in situ (CIS) have a considerably higher risk of progression and metastases. Thus, prospective clinical trials should consider the different risks associated with different prognostic groups and stratify patients accordingly. The Dutch Cooperative Trial evaluated mitomycin versus BCG-Tice versus BCG-RIVM in 469 patients with pTA/pT1 carcinoma and CIS of the urinary bladder after TUR. Of 437 evaluable patients, 50 had CIS, 254 had pTA tumors, and 133 had pT1 tumors. No statistical differences were observed in toxicity between the two strains of BCG, but local and systemic side effects were more frequent in the BCG groups versus the mitomycin group. No differences in response rate were observed between the 3 treatment groups in patients with CIS. A statistically significant difference in favor of mitomycin was seen, however, in patients with papillary tumors. Mitomycin and BCG-RIVM were equally effective, and mitomycin proved significantly more effective than BCG-Tice. No significant difference in efficacy was observed between the two strains of BCG. Disease recurrence during the study in patients with papillary tumors was seen in 43% of mitomycin-treated patients, 64% of the BCG-Tice group, and 46% of patients treated with BCG-Rivm. However, a subgroup analysis for time to first recurrence for pTa versus pT1 and for grade 1 and 2 versus grade 3 papillary tumors showed no statistically significant between-group difference. Further studies comparing identical regimens, doses, and patient groups are needed to define more clearly which patient groups and tumors are more likely to respond to intravesical therapy. PMID- 8727806 TI - Intravesical chemotherapy versus immunotherapy for superficial bladder cancer. AB - The decision to treat superficial bladder cancer with intravesical therapy should be predicated primarily on disease stage and grade as well as the patient's clinical history. Once the decision to proceed with intravesical therapy has been made, the clinician must select the appropriate agent. Several agents are available and the choice of which agent to use should be based on careful consideration of the potential benefit of a given drug versus its inherent risk of complications. The first drug to be administered intravesically, thiotepa is an alkylating agent used as first-line treatment for low-grade lesions; it has limited use against higher-grade tumors or carcinoma in situ. In addition, the low molecular weight of thiotepa results in significant systemic absorption, which often results in myelosuppression. Mitomycin, also an alkylating agent, has shown significant activity as both first-line therapy and in patients with recurrent disease. Unlike thiotepa, mitomycin has a relatively high molecular weight, and the incidence of significant bladder absorption and systemic side effects is low. Doxorubicin, which also possesses a high molecular weight, is used intravesically against superficial bladder cancer more frequently in Europe and Japan than in the United States. Immunotherapy with bacille Calmette-Guerin is the treatment of choice for carcinoma in situ and high-grade T1 lesions. It is associated with the highest incidence of both minor and major adverse reactions, however, and its use should be tempered by its potential toxicity. PMID- 8727807 TI - Pathology and its importance in evaluating outcome in patients with superficial bladder cancer. AB - Subtle cytologic and histologic nuances have a major impact on diagnosis and, consequently, on therapy for superficial bladder cancer. Therefore, the urologist and the pathologist must carefully assess all clinical findings before a course of treatment can be determined. The urologist must advise the pathologist of all the circumstances surrounding a biopsy--whether its purpose is for preliminary clinical impression or diagnosis, the patient's recent treatment history, the availability of previous biopsy specimens for comparison, a thorough history of treatments that may induce characteristic cytologic changes that might lead to misdiagnosis, and alternate diagnostic possibilities drawn from initial pathology and treatment history. Armed with this information, the task of the pathologist is to provide as much data as possible regarding tumor histopathology from the biopsy specimens. Thus, establishment of a close working relationship between the urologist and the pathologist is an important tool for (1) initially characterizing superficial bladder cancer, which is essential in determining an appropriate course of treatment, and (2) accurately evaluating follow-up biopsies to determine the effectiveness of that treatment. PMID- 8727808 TI - Intravesical chemotherapy for superficial bladder tumors category Ta/T1: who should be treated and how? AB - Superficial bladder cancer is the presenting feature in 75% to 80% of patients with newly diagnosed bladder carcinoma. Treatment goals are (1) to eradicate existing disease, (2) to provide prophylaxis against recurrence, and (3) to prohibit muscular invasion and metastases to lymph nodes. First-line treatment for stages Ta and T1 disease is transurethral resection (TUR). Recurrence rates with surgery alone, however, range from 40% to 80%. Study results show a statistically significant benefit in treatment results over those observed with TUR alone when adjuvant intravesical instillation of cytotoxic agents or immunomodulators is added. Patient characteristics are important factors in determining which patients might benefit from adjuvant treatment. Based on several trials reported by the European Organization for the Research and Treatment of Cancer Genitourinary Group, the following conclusions can be drawn: (1) patients with < 1 recurrence/year or with a primary solitary bladder tumor category Ta G2-3 or T1 G1-3 may benefit from a single, early instillation of a cytotoxic agent, (2) solitary Ta G1 lesions may be treated with TUR alone, (3) adjuvant therapy with cytotoxic agents should be instituted as soon as possible after TUR (this is not feasible with immunotherapy), (4) for patients treated within 6 hours after or on the day of TUR, a 6-month course of therapy is sufficient, whereas a 12-month course provides better results for patients in whom such early intravesical instillation is not feasible. PMID- 8727809 TI - Nursing implications in the management of superficial bladder cancer. AB - Intravesical therapy is preferred over systemic treatment for superficial bladder cancer. Four agents commonly used are thiotepa, doxorubicin, mitomycin, and bacille Calmette-Guerin. A thorough understanding of the unique methods involved in the instillation of these drugs and the special patient-management procedures associated with their use is critical for nursing personnel. Occupational Safety and Health Administration guidelines exist to ensure safe handling of intravesical agents. Nurses are urged to take precautions to limit their exposure to these potentially hazardous drugs. For example, bacille Calmette-Guerin, a live virus, represents a biohazard and extreme caution is required in all aspects of handling, particularly in its disposal. Because of the potential complications associated with intravesical therapy, a significant responsibility of the nurses who administer it is patient education. Patients need to know what to expect during the treatment process as well as how to recognize the signs and symptoms of complications. In addition to gaining a thorough knowledge of the techniques involved in the intravesical treatment of superficial bladder cancer, nurses can also serve their patients better by using sensitive to the unique psychosocial problems they face. PMID- 8727810 TI - Comparison of surgical treatments for trigeminal neuralgia: reevaluation of radiofrequency rhizotomy. AB - In this study, we reevaluate the results of radiofrequency rhizotomy and review the effectiveness of other surgical procedures for the treatment of trigeminal neuralgia. Five hundred patients with trigeminal neuralgia underwent radiofrequency rhizotomy at the University of Cincinnati Medical Center, Cincinnati, OH, between 1981 and 1986. Their results are compared with those of patients reported in the literature who underwent radiofrequency rhizotomy (6205 patients), glycerol rhizotomy (1217 patients), balloon compression (759 patients), microvascular decompression (MVD) (1417 patients), and partial trigeminal rhizotomy (250 patients). Comparisons were based on the following outcome parameters: technical success, pain relief and recurrence, facial numbness, dysesthesia, corneal anesthesia, keratitis, trigeminal motor dysfunction, permanent cranial nerve deficit, intracranial hemorrhage or infarction, perioperative morbidity, and perioperative mortality. We found that MVD had the lowest rate of technical success. Radiofrequency rhizotomy and MVD had the highest rates of initial pain relief and the lowest rates of pain recurrence. Glycerol rhizotomy had the highest rate of pain recurrence. Balloon compression had the highest rate of trigeminal motor dysfunction. Balloon compression and MVD had the lowest rates of corneal anesthesia or keratitis. MVD had the lowest rates of facial numbness and dysesthesia. All percutaneous procedures had similar rates of dysesthesia. Posterior fossa exploration had the highest rates of permanent cranial nerve deficit, intracranial hemorrhage or infarction, and perioperative morbidity and mortality. On the basis of our experience and a review of the literature, we conclude the following: 1) percutaneous techniques and posterior fossa exploration offer advantages and disadvantages, 2) radiofrequency rhizotomy is the procedure of choice for most patients undergoing first surgical treatments, and 3) MVD is recommended for healthy patients who have isolated pain in the first ophthalmic trigeminal division or in all three trigeminal divisions and patients who desire no sensory deficit. PMID- 8727811 TI - Variations in the natural history and survival of patients with supratentorial low-grade astrocytomas. AB - Data from 55 consecutive patients with low-grade astrocytomas treated between 1982 and 1990 were analyzed to determine specific outcome factors, including time to recurrence, incidence of anaplastic transformation, and survival. Gender, type of symptoms, contrast enhancement, and timing of radiation therapy were not significant in determining outcome. Patients who had symptoms for > 2 years and underwent gross-total resection of the tumor, with age as a continuous variable, were associated with significantly longer time to recurrence and survival. Within the population of patients with low-grade astrocytomas, patients with chronic epilepsy clearly had the best prognoses. There were no tumor recurrences or deaths in 27 patients with chronic epilepsy, regardless of the extent of surgery and without the use of radiotherapy. Ten-year survival was 100% for 31 patients who underwent gross-total tumor resection, regardless of the length of preoperative symptoms. Immediate postoperative radiotherapy did not prolong the time to recurrence, reduce the incidence of transition to more malignant tumors at recurrence, or increase the length of survival when compared with delayed radiotherapy. Because recurrence with a high-grade lesion caused 92% of the mortality in our series, the benefit in patients who underwent aggressive surgery seems to result from a significant decrease in the risk of recurrence when compared with patients who underwent anything less than gross-total resection. Our data also suggest that variability in the natural history of low-grade astrocytomas has a strong influence in determining survival and that tumors associated with chronic epilepsy are much less likely to become more malignant over time. PMID- 8727812 TI - The neuroimaging and clinical spectrum of neurofibromatosis 2. AB - Neurofibromatosis 2 (NF2) is an autosomal dominant disease predisposing to multiple tumors of the central and peripheral nervous system. Bilateral vestibular schwannomas are the hallmark of the disease. To define the clinical spectrum of the disease, we performed gadolinium-enhanced magnetic resonance imaging of the brain and spine as well as neurological, dermatological, and ocular examinations in 48 patients with NF2 diagnosed with the National Institutes of Health diagnostic criteria. Patients were ascertained from patient workshops and publications and from referral as a result of vestibular schwannoma surgery. Vestibular schwannomas were found in 46 patients (96%, 43 bilateral and 3 unilateral), spinal tumors were found in 43 (90%), posterior subcapsular cataracts were found in 30 (63%), meningiomas were found in 28 (58%), and trigeminal schwannomas were found in 14 (29%). The presenting symptoms included hearing loss or tinnitus in 15 patients (31%), multiple or nonspecific symptoms in 15 (31%), skin tumors in 12 (25%), and ocular symptoms in 6 (13%). When the complete spine was imaged, spinal tumors were more common in patients with NF2 than has previously been reported. This is a noteworthy finding, because spinal tumors are a major cause of NF2 morbidity and mortality. PMID- 8727813 TI - The incidence of surgical complications is similar in good and poor grade patients undergoing repair of ruptured anterior circulation aneurysms: a retrospective review of 355 patients. AB - To determine how clinical grade after subarachnoid hemorrhage impacts operative characteristics and the incidence of intra- and postoperative surgical complications, we retrospectively compared the surgical management of all good grade (n = 224) and poor grade (n = 131) patients who suffered ruptured anterior circulation aneurysms between 1983 and 1993. The majority of good grade (74.2%) and poor grade (89.8%) patients underwent surgery < 3 days after subarachnoid hemorrhage. The results in this series demonstrate that severe cerebral swelling, often secondary to intracerebral hemorrhage, was significantly more frequent in poor grade patients. The incidence of complications, such as failure to occlude the aneurysm, major vessel occlusion, intraoperative aneurysm rupture, or surgical contusion, however, was similar in poor grade and good grade patients. We conclude, therefore, that except for severe cerebral swelling associated with intracerebral hemorrhage, the risk of surgical complications is similar in good and poor grade patients undergoing surgical repair of ruptured anterior circulation aneurysms. PMID- 8727814 TI - The relationship of blood velocity as measured by transcranial doppler ultrasonography to cerebral blood flow as determined by stable xenon computed tomographic studies after aneurysmal subarachnoid hemorrhage. AB - Transcranial doppler (TCD) ultrasonography is often used to guide the management of patients with subarachnoid hemorrhage (SAH). However, the correlation between increased blood velocity as measured by TCD ultrasonography and angiographic vasospasm was established before the routine use of hypervolemia/hemodilution and administration of nimodipine and did not address blood flow. The relationship of blood velocity as measured by TCD ultrasonography and local cerebral blood flow (LCBF) in SAH managed with these modalities is unknown. Patients presenting with aneurysmal SAH between January 1992 and September 1993 who underwent TCD ultrasonography and xenon computed tomographic (Xe/CT) LCBF studies within 12 hours were retrospectively studied. Fifty patients underwent a total of 94 paired studies, encompassing 709 vascular territories. All were treated with nimodipine and hypervolemia/hemodilution. Hematocrit, blood pressure, and partial carbon dioxide pressure were similar at the time of TCD ultrasonography and Xe/CT measurement of LCBF. When LCBF in the middle cerebral artery (MCA) was < or = 31 ml/100 g/min, the corresponding peak systolic velocity measured by TCD ultrasonography was 119 cm/s, whereas those > 31 ml/100 g/min had a velocity of 169 cm/s (P = 0.006). High LCBF was associated with high velocity in all vascular territories, reaching significance in all but the internal carotid artery. At the time of each study, 41 neurological examinations were focal and 53 were nonfocal. The Xe/CT measurement of LCBF in the MCA contralateral to a deficit was significantly less than in territories without corresponding clinical deficits (P = 0.01), whereas peak systolic velocities in the MCA were not significantly different (P = 0.71). Territories with increases in blood velocity in the MCA of > 50 cm/s/24 h did not have statistically different LCBF (P = 0.183). Our results suggest that increased blood velocity revealed by TCD ultrasonography correlates with increased LCBF and not with ischemia. No difference in LCBF was found in territories with and without rapid increases in blood velocity in the MCA. Furthermore, although focal neurological deficits corresponded with decreased contralateral LCBF in the MCA, increased velocity did not correlate with neurological findings. Therapeutic decisions based solely on blood velocity revealed by TCD ultrasonography might be inappropriate and potentially harmful. Xe/CT studies of LCBF are useful in guiding the management of SAH. PMID- 8727816 TI - Expression of angiogenic factors and structural proteins in central nervous system vascular malformations. AB - Little is known of the molecular mechanisms mediating the genesis and subsequent biological behavior of central nervous system vascular malformations. The role of angiogenic and permeability-inducing factors in the pathogenesis of these lesions has not bee previously explored. In this study, we subject specimens from 12 cases of excised vascular malformation to a battery of immunostaining for vascular endothelial growth factor, basic fibroblast growth factor, and selected structural and matrix proteins. The lesions consisted of seven arteriovenous malformations (AVMs), including one angiographically occult AVM, one arterialized vein from a dural AVM, and five cavernous malformations (CMs). Vascular endothelial growth factor was expressed by all lesions and was localized predominantly in the subendothelial layer and in perivascular spaces. Four of seven AVMs and four of five CMs demonstrated faint basic fibroblast growth factor expression that was localized to the media of AVM vessels and the subendothelial layer and intercavernous matrix of CMs. This pattern of angiogenic factor immunostaining was correlated with the expression of structural and matrix proteins in the same lesions. Laminin was not expressed in any of the CMs, confirming previous reports from our laboratory. By contrast, fibronectin expression was more prominent in CMs than in AVMs. Collagen Type IV and alpha smooth muscle actin expression occurred in every lesion. We conclude that angiogenic growth factors are expressed in all types of vascular malformations of the central nervous system. The pattern of expression suggests diffuse activation of angiogenesis without specific relation to individual vessel types or recent clinical behavior. Defining the role of angiogenesis in vascular malformations might provide insight into their pathogenesis and suggest novel strategies for modification of their behavior. PMID- 8727815 TI - Gamma knife radiosurgery for arteriovenous malformations: long-term follow-up results focusing on complications occurring more than 5 years after irradiation. AB - The detailed long-term follow-up results of 40 patients treated for cerebral arteriovenous malformations with gamma knife radiosurgery are presented, with special reference to postradiosurgical complications that can develop many years after irradiation. The follow-up period after radiosurgery was 54 to 205 months, excluding one mortality, with a mean and a median of 106 and 97 months, respectively. One patient (2.5%) has, to date, refused all neuroimaging follow-up examinations. Complete nidus obliteration was angiographically confirmed in 26 patients (65%) between 1 and 5 years after radiosurgery. In the remaining 13 patients (32.5%), although significant shrinkage of each nidus was angiographically demonstrated, complete obliteration was not attained during a 3- to 7-year period of follow-up after radiosurgery. Among these 13 patients, 1 underwent surgical extirpation of the nidus and 5 underwent second courses of gamma knife radiosurgery between 3 and 6 years after initial treatment; in 3 of the 5 patients, complete nidus obliteration was angiographically confirmed between 1 and 3 years after the second course of radiosurgery. There were no radiation- or arteriovenous malformation-related mortalities. However, we did experience one angiography-related mortality. We also experienced one morbidity (probably caused by hemorrhagic stroke), which developed 5 years after 2-year postradiosurgical angiography had demonstrated complete obliteration, and three radiation-related morbidities, two of which (hemiparkinsonian syndrome and visual field disturbances caused by delayed cyst formation) occurred 5.5 and 7 years, respectively, after irradiation. Furthermore, we observed another two patients who, although asymptomatic to date, showed delayed cyst formation on magnetic resonance imaging 5 and 10 years after irradiation, respectively. In total, 3 (23%) of 13 patients who underwent computed tomography and/or magnetic resonance imaging more than 5 years after radiosurgery showed delayed cyst formation. In conclusion, long-term follow-up, particularly with the use of neuroimaging techniques, is necessary even after the treatment goal has been achieved. PMID- 8727817 TI - Neurosurgical management of thoracic and lumbar vertebral osteomyelitis and discitis in adults: a review of 43 consecutive surgically treated patients. AB - We report 43 consecutive surgically treated patients with pyogenic (37 patients) and tuberculous (6 patients) osteomyelitis of the thoracic and lumbar spine encountered within an 8-year period, including 1 with late recurrence after 15 months. There were 24 men and 18 women, ranging in age from 21 to 83 years. Twenty-six patients were in poor general condition because of associated illnesses, especially diabetes mellitus. Disease occurred at the thoracic level in 19 patients and on the lumbar spine in 24. After diagnosis, five patients were merely treated by posterior decompression; three of them, however, required further surgery for recurrent infection, spinal instability, and secondary neurological impairment. They are included in the 40 patients who underwent combined posterior debridement and internal fixation with transpedicular screw rod systems. Autologous interbody bone grafting was performed simultaneously in 18 patients and in a second stage operation in 21 patients. One of them (tuberculous) experienced early recurrence and required anterior fusion. In two patients, methylmethacrylate packing was used for spine reconstruction; one of them had a late recurrence. Of the 26 patients with preoperative marked or severe neurological deficit (Frankel Grades A, 2 patients; B, 1 patient; C, 17 patients; and D, 6 patients), 23 (88%) had significant improvement of one grade (15 patients) or more (8 patients). There were no permanent complications. However, intensive care treatment was necessary in 20 of the 26 patients in reduced general condition (mean age, 72 yr). Two patients required further surgery because of postoperative epidural hematoma and pedicle screw malpositioning. In conclusion, most patients with thoracic and lumbar osteomyelitis can be successfully treated by combined debridement and internal fixation using only a posterior approach. Autogenous interbody bone grafting can be simultaneously performed and allows early mobilization of the patient. PMID- 8727818 TI - Continuous intrathecal baclofen infusion for symptomatic generalized dystonia. AB - Five patients with generalized dystonia who were refractory to oral medications were treated by continuous intrathecal baclofen infusion. Dystonia was related to cerebral palsy in three patients and to Hallervorden-Spatz disease in two. Responsiveness to intrathecally administered baclofen was evaluated after bolus injections in one patient and during continuous infusions via an external micropump in four. Patients who responded to trial injections were subsequently implanted with a programmable pump for continuous infusion of baclofen. Dystonia in the three patients were cerebral palsy was substantially improved by continuous intrathecal baclofen infusion in doses of 500 to 800 micrograms/d. Benefit has persisted for > 19 months of continuous infusion. Dystonia in the two patients with Hallervorden-Spatz disease was not improved, although the screening trial was limited by side effects in one patient and by meningitis in the other. We conclude that continuous intrathecal baclofen infusion is beneficial therapy for some patients with generalized dystonia and that additional investigations are indicated. PMID- 8727819 TI - Dysesthetic pain in patients with syringomyelia. AB - Dysesthethic pain is a common complaint of patients with syringomyelia, traumatic paraplegia, and various myelopathic conditions. Because cavitary lesions of the spinal cord can be defined with good resolution by magnetic resonance imaging, syringomyelia provides a potential model for examining anatomic correlates of central pain. In this study, a syndrome of segmental dysesthesias, characterized by burning pain, hyperesthesia, and a variable incidence of trophic changes, was described by 51 of 137 patients (37%) with syringomyelia at the time of clinical presentation. Complete magnetic resonance scans, including axial images, demonstrated extension of the syrinx into the dorsolateral quadrant of the spinal cord on the same side and at the level of pain in 43 of 51 patients (84%). Surgical treatment of syringomyelia resulted in the relief or improvement of dysesthetic pain in 22 of 37 patients (59%), but 15 patients (41%) reported no improvement or an intensification of pain despite collapse of the syrinx. Postoperative dysesthetic pain was often a disabling complaint that responded poorly to medical therapy, including analgesics, sedatives, antiepileptics, antispasmodics, and anti-inflammatory agents. In most cases, there was a gradual improvement of symptoms, although six patients continued to complain of pain 24 to 74 months postoperatively. Prompt but transient relief was achieved in two of two patients with regional sympathetic blocks, and prolonged relief was achieved in one patient by stellate ganglionectomy. We conclude that painful dysesthesias can be caused by a disturbance of pain-modulating centers in the dorsolateral quadrant of the spinal cord and have certain causalgia-like features that respond in an unpredictable way to surgical collapse of the syrinx. PMID- 8727820 TI - Selective occlusion of basilar artery aneurysms using controlled detachable coils: report of 35 cases. AB - The development of new devices, especially controlled detachable coils, has made the endovascular approach one of the modalities for the treatment of intracranial aneurysms. We describe the treatment and present the results of 35 patients treated by selective occlusion of basilar artery aneurysms in our department during a period of 2 years (November 1992-November 1994). This period of time was chosen to analyze a homogeneous population treated since the introduction of controlled detachable coils and also to be able to have as many follow-up angiographic controls of the treated aneurysms as possible. The clinical presentation was subarachnoid hemorrhage in 32 patients and transient ischemic attack in 1 patient. In another two patients, the aneurysms were incidentally discovered. The majority of the aneurysms were berry aneurysms. The aneurysms were located at the basilar bifurcation (23 patients), at the basilar tip between the posterior cerebral artery and the superior cerebellar artery (5 patients), on the basilar trunk (3 patients), and at the vertebrobasilar junction (4 patients). Endovascular treatment using coils was achieved in 34 patients, using Guglielmi detachable coils (Target Therapeutics, San Jose, CA) in 29 patients and mechanical detachable spirals (Balt, Montmorency, France) in 5 patients. One patient died during the positioning of the first coil into the aneurysmal sac. Twenty-five of 35 aneurysms (73.5%) were completely occluded. Nine aneurysms (26.5%) were only partially (> 90%) occluded. No subsequent bleeding occurred during the follow-up period. Two patients treated in the acute phase of subarachnoid hemorrhage died days or weeks after endovascular treatment because of complications related to the natural history of subarachnoid hemorrhage (vasospasm in one patient and pulmonary complications in the other). In three patients, clotting occurred during the endovascular procedure. In all three patients, occlusion of the aneurysmal sac was achieved despite clotting. Urokinase was administered to two of the three patients. In the remaining patient, no fibrinolytic therapy was initiated. The clinical outcomes were excellent for all three patients. In this study, the morbidity-mortality rate of the endovascular technique is low (3%). If we include complications related to the subarachnoid bleeding, the morbidity-mortality rate remains low (8.8%) Regarding basilar artery aneurysms, endovascular treatment (selective occlusion by controlled detachable coils) is now useful for some patients, especially those with small aneurysms. However, long-term anatomic follow-up is needed to accurately evaluate the role of this treatment modality in the management of basilar aneurysms. PMID- 8727821 TI - Aneurysm clipping after endovascular treatment with coils: a report of eight patients. AB - Between January 1990 and December 1994, patients with subarachnoid hemorrhage related to ruptured aneurysms who were referred to our institution were treated by neurosurgical and neuroradiological teams. In each patient, the respective indications for neurosurgical or endovascular treatment were discussed, taking into consideration patients' age and the morphological and topographical aneurysm features. We report eight cases of patients with subarachnoid hemorrhage who underwent operations after primary endovascular procedures (Hunt and Hess scores III, IV, and V). The indications for surgical treatment were as follows. First, deliberate partial occlusion of the aneurysm (two aneurysms of the internal carotid artery and one aneurysm of the anterior communicating artery) was performed to obtain only partial clotting of the aneurysm sac by free coils. However, this procedure was discontinued in favor of the use of Guglielmi detachable coils. The second indication was partial occlusion after an endovascular procedure (two aneurysms of the middle cerebral artery and one internal carotid artery aneurysm). The third indication was re-expansion of the aneurysm 1 year after the endovascular treatment (one middle cerebral artery aneurysm). The final indication was secondary rupture of the aneurysm sac and false aneurysm around the migrating coil (one aneurysm of the pericallosal artery). During surgery, the aneurysm sac appeared translucent. The coils bulged out and stretched the aneurysm sac. One ruptured the membrane leading to a subarachnoid hemorrhage during the endovascular procedure. No hemorrhage occurred during the surgical clipping. Aneurysm obliteration was easily performed, especially when the packing was partial, but was very difficult when the complete aneurysm closure led to a stenosis of the parent vessel. A giant sylvian aneurysm rest, visible only with angiography, was left untreated. This series illustrates an original experience, which led us to conclude that aneurysm surgery with coils in place is not as difficult as is often thought. PMID- 8727822 TI - Somatosensory evoked potential phase reversal and direct motor cortex stimulation during surgery in and around the central region. AB - In 99 patients with mass lesions in and around the central region, the central sulcus was intraoperatively localized with the use of somatosensory evoked potential (SEP) phase reversal. In 33 of these patients, the motor cortex was directly stimulated and electromyographic responses were recorded from the forearm flexor, thenar, and hypothenar muscles. An additional 25 patients, with subcortical lesions or lesions directly located at the pyramidal tract, were continuously monitored during surgery by motor evoked potentials (MEPs). An exact determination of the central sulcus and tumor localization was possible in all patients; a phase reversal was obtained in 90.8% of the patients, and localization was possible as a result of anatomic structures and the loss of N20 or P20 of SEPs in the other 9.2%. MEPs were obtained in 32 of 33 patients and also in all 25 patients who underwent MEP monitoring at the beginning of tumor removal. From this study, it can be concluded that the combination of SEP phase reversal and modified electrical cortex stimulation is compatible with general anesthesia, although anesthesia was not systematically controlled according to a protocol. Although this study demonstrates that the combined SEP/MEP technique was feasible, it is not yet possible to demonstrate benefit in improving the outcome of patients. Concerning the safety of stimulation, the exact localization of the central sulcus by the noninvasive SEP method, compared with direct electrical stimulation, offered more safety for the patient. The modified technique of direct motor cortex stimulation needed much less charge density than did the common technique. The recording of electromyographic responses instead of movements made objective documentation possible, and the analysis of amplitudes and latencies will supply quantitative information about the motor system. PMID- 8727823 TI - Anatomic study of anterior frontal cortical bridging veins with special reference to the frontopolar vein. AB - Anatomic variations of the frontopolar vein were investigated in 21 cadaver brains to improve the preservation rate of this vein in interhemispheric surgery for anterior communicating aneurysms. Most of the frontopolar vein has been considered to drain the outer convex side of the frontal lobe, but in reality, the area of its venous drainage was found to cover a large part of the frontal lobe, including its medial and basal surfaces. This observation suggests that sacrifice of the vein during surgery carries a risk of venous infarction. Therefore, care must be taken to not injure the vein during surgery. The mean distance between the frontopolar vein and the most anterior point of the frontal lobe was 31.1 mm. Although the vein became smaller close to the frontal tip, the mean diameter of the vein was 1.9 mm. Morphologically, approximately two-thirds of the frontopolar veins was found to have a "main trunk." The frontopolar vein can be spared during surgery by using a flexible surgical technique incorporating vein dissection or additional craniotomy. PMID- 8727824 TI - Reduction of extraneural scarring by ADCON-T/N after surgical intervention. AB - The effects of ADCON-T/N (Gliatech, Inc., Cleveland, OH), a carbohydrate polymer gel, on peripheral nerve scarring and regeneration were studied in rodents undergoing three types of surgical intervention. Procedure I involved external neurolysis of the sciatic nerve from surrounding tissues and separation of its tibial and peroneal components. Procedure II involved the addition of an abrasive injury. Procedure III involved transection and suture anastomosis of the tibial component. ADCON-T/N or a control gel was locally applied in a blind fashion. Additional animals received no gel, as a further control. Animals underwent second operations 4 weeks after Procedures I and II and 6 weeks after Procedure III. The surgical sites were evaluated using a numerical grading scheme to assess wound healing, sciatic nerve adherence to surrounding tissues, and separability of its tibial and peroneal components. Animals receiving ADCON-T/N demonstrated reduced nerve adherence to surrounding tissues and enhanced separability of the tibial and peroneal components, compared with animals receiving control gel or no gel. Quantitative histological analysis revealed a statistically significant reduction in the amount of dense scar tissue surrounding nerves treated with ADCON-T/N. No evidence of nerve toxicity caused by ADCON-T/N was noted. Counts of regenerating myelinated axons in animals undergoing nerve transection and suture repair did not statistically differ in treated and untreated animals. In conclusion, ADCON-T/N seems to be both safe and effective in reducing extraneural scar formation after peripheral nerve surgery and local trauma. PMID- 8727825 TI - Enhanced delivery of boronophenylalanine for neutron capture therapy by means of intracarotid injection and blood-brain barrier disruption. AB - There has been increasing interest in the possible use of boronophenylalanine as a capture agent for boron neutron capture therapy of brain tumors. The purpose of the present study was to determine whether the uptake of boronophenylalanine in F98 glioma-bearing rats could be enhanced by means of intracarotid (i.c.) injection with or without blood-brain barrier disruption (BBB-D). Glioma cells (10(5)) were stereotactically implanted into the right cerebral hemisphere of Fischer rats, and 12 days later, BBB-D was performed by infusing 25% mannitol (1.373 mOsmol/ml) into the right carotid artery and then immediately injecting L boronophenylalanine (300 mg/kg of body weight) intracarotidly. The animals were killed 0.5, 1, 2.5, and 4 hours later, and the brains were removed for boron determination by direct current plasma atomic emission spectroscopy. BBB-D was assessed by the intravenous injection of Evans blue or horseradish peroxidase, and the barrier-disrupted hemispheres and tumors showed intense staining with each. The mean tumor boron concentration after i.c. injection and BBB-D was 34.8 +/- 6.8 micrograms/g at 2.5 hours compared with 20.3 +/- 6.2 micrograms/g after i.c. injection without BBB-D and 10.7 +/- 0.7 micrograms/g after intravenous injection. No significant differences in boron concentration in muscle, skin, and eye were observed among the different groups. Boron concentrations in the ipsilateral, disrupted hemisphere increased transiently but rapidly returned to background levels by 2.5 hours after BBB-D. The tumor:brain and tumor:blood ratios were 5.2 and 5.6, respectively, compared to 3.2 and 2.1 for intravenous injection groups at 2.5 hours. The present study is the first to show that BBB-D combined with i.c. injection can enhance the tumor uptake of boron compounds for boron neutron capture therapy. PMID- 8727826 TI - Axon and ganglion cell injury in rabbits after percutaneous trigeminal balloon compression. AB - New Zealand white rabbits were used to determine whether the changes in the Vth cranial nerve sensory root after compression were associated with the loss of a specific subclass of Vth cranial nerve ganglion cells, the disappearance of a distinct subset of primary afferent terminals in Vth cranial nerve nucleus caudalis, and/or injury to a specific axonal fiber type. There was no significant difference in the size of surviving ganglion cells after Vth cranial nerve compression, as measured 2 to 3 months after injury (P > 0.5, n = 4). Densitometric analysis of the nerves of rabbits that survived > 2 months after compression showed no significant difference in the immunoreactivity of substance P and calcitonin gene-reactive protein between compressed and control sides (P > 0.1, n = 4). Fink-Heimer staining of the Vth cranial nerve subnucleus caudalis revealed that transganglionic degeneration was most dense in the deeper layers, which are the sites of termination of large myelinated fibers. Ultrastructural evaluation of the type of myelinated axons injured by Vth cranial nerve compression in rabbits killed 7, 14, 37, and 270 days after injury was studied, and morphometric analysis was performed. The frequency distribution of axon diameters was significantly different for injured and control areas. The injured areas had higher ratios of small (< 3-microns diameter) to large-diameter axons compared to control distribution. These data indicate that balloon compression results in loss of fibers from the Vth cranial nerve sensory root and extensive transganglionic degeneration in the Vth cranial nerve brain stem complex. Cell size measurements and immunocytochemical data suggest that there is no specific loss of small ganglion cells or fine-caliber primary afferents. These experiments suggest that balloon compression relieves trigeminal pain by injuring the myelinated axons involved in the sensory trigger to the pain. PMID- 8727827 TI - A biomathematical model of intracranial arteriovenous malformations based on electrical network analysis: theory and hemodynamics. AB - Hemodynamics play a significant role in the propensity of intracranial arteriovenous malformations (AVMs) to hemorrhage and in influencing both therapeutic strategies and their complications. AVM hemodynamics are difficult to quantitate, particularly within or in close proximity to the nidus. Biomathematical models represent a theoretical method of investigating AVM hemodynamics but currently provide limited information because of the simplicity of simulated anatomic and physiological characteristics in available models. Our purpose was to develop a new detailed biomathematical model in which the morphological, biophysical, and hemodynamic characteristics of an intracranial AVM are replicated more faithfully. The technique of electrical network analysis was used to construct the biomathematical AVM model to provide an accurate rendering of transnidal and intranidal hemodynamics. The model represented a complex, noncompartmentalized AVM with 4 arterial feeders (with simulated pial and transdural supply), 2 draining veins, and a nidus consisting of 28 interconnecting plexiform and fistulous components. Simulated vessel radii were defined as observed in human AVMs. Common values were assigned for normal systemic arterial pressure, arterial feeder pressures, draining vein pressures, and central venous pressure. Using an electrical analogy of Ohm's law, flow was determined based on Poiseuille's law given the aforementioned pressures and resistances of each nidus vessel. Circuit analysis of the AVM vasculature based on the conservation of flow and voltage revealed the flow rate through each vessel in the AVM network. Once the flow rate was established, the velocity, the intravascular pressure gradient, and the wall shear stress were determined. Total volumetric flow through the AVM was 814 ml/min. Hemodynamic analysis of the AVM showed increased flow rate, flow velocity, and wall shear stress through the fistulous component. The intranidal flow rate varied from 5.5 to 57.0 ml/min with and average of 31.3 ml/min for the plexiform vessels and from 595.1 to 640.1 ml/min with an average of 617.6 ml/min for the fistulous component. The blood flow velocity through the AVM nidus ranged from 11.7 to 121.1 cm/s with an average of 66.4 cm/s for the plexiform vessels and from 446.9 to 480 dyne/cm2 with an average of 463.5 dyne/cm2 for the fistulous component. The wall shear stress ranged in magnitude from 33.2 to 342.1 dyne/cm2 with an average of 187.7 dyne/cm2 for the plexiform vessels and from 315.9 to 339.7 cm/s with an average of 327.8 cm/s for the fistulous component. The described novel biomathematical model characterizes the transnidal and intranidal hemodynamics of an intracranial AVM more accurately than was possible previously. This model should serve as a useful research tool for further theoretical investigations of intracranial AVMs and their hemodynamic sequelae. PMID- 8727828 TI - The development of neurological surgery at the Salpetriere Hospital. AB - The development of neurological surgery at the Salpetriere Hospital is traced from its beginning, at the end of the 19th century, with reference to the historical aspect of the hospital itself, going back over 300 years. Early leading neurological surgeons and their achievements are described, and a summary of the present department of neurosurgery and its future plans is presented. PMID- 8727829 TI - Early approaches to brain tumors. PMID- 8727830 TI - Dorsal brain stem lipomas: case report. AB - Intracranial lipomas are uncommon and rarely symptomatic lesions accounting for 0.06 to 0.46% of intracranial lesions. The management of symptomatic dorsal brain stem lipomas was once limited to cerebrospinal fluid diversion, but with recent advances in microsurgery, they now may be directly treated. We report three patients with dorsal brain stem lipomas, two of which involved the quadrigeminal cistern and one of which was in the cisterna magna region. Antenatal documentation by ultrasound examination in one patient represents the first reported in utero diagnosis of quadrigeminal cistern lipoma. Computed tomographic and magnetic resonance imaging scans were diagnostic. The surgical experience in two symptomatic patients is discussed. Microsurgical decompression was performed in each without neurological deficit, and clinical symptoms postoperatively subsided. No patient required a permanent cerebrospinal fluid shunt. The management of symptomatic dorsal brain stem lipomas is discussed, and an algorithm is proposed. PMID- 8727831 TI - T-cell type primary spinal intramedullary lymphoma associated with human T-cell lymphotropic virus type I after a renal transplant: case report. AB - We present the first report of primary spinal intramedullary lymphoma of the T cell type associated with human T-cell lymphotropic virus Type I infection in a 52-year-old woman who had undergone a renal transplant. The pathogenesis of spinal cord T-cell lymphoma is discussed. PMID- 8727832 TI - An unusual cause of proptosis: orbital solitary fibrous tumor: case report. AB - Although solitary fibrous tumors are well-recognized tumors, they have only recently been described in the orbit. Although histopathological awareness of the lesion has been heightened recently because of the discovery of CD-34 immunoreactivity in solitary fibrous tumors, including orbital lesions, it remains unrecognized clinically as a cause of unilateral proptosis. Clinical awareness of the lesion is important, because benign and malignant forms of the tumor occur. The malignant forms pursue an aggressive course manifested by local invasion, recurrent growth, or metastases. The benign lesions are histologically banal and are cured by surgical excision. This case report describes unilateral proptosis caused by an orbital solitary fibrous tumor, which occurred in a 43 year-old woman. The tumor had a benign histomorphology and clinical course, with surgical excision being curative. In reviewing the literature on orbital solitary fibrous tumors, clinical awareness of the tumor as a cause of unilateral proptosis is emphasized. PMID- 8727833 TI - Sagittal sinus occlusion, caused by an overlying depressed cranial fracture, presenting with late signs and symptoms of intracranial hypertension: case report. AB - A case of delayed signs of intracranial hypertension after open depressed cranial fracture occluding the superior sagittal sinus is reported. Elevating depressed fractures overlying a cranial venous sinus is hazardous. Conservative management of the intracranial hypertension, including repeated lumbar punctures, led to an unimpaired outcome. The options of management of delayed problems caused by traumatic venous occlusion are discussed. PMID- 8727834 TI - Middle fossa cyst presenting as a delayed complication of temporal lobectomy: case report. AB - Symptomatic middle fossa cysts have not previously been described as complications of temporal lobectomy that has been performed to control complex partial seizures. A patient is described who developed a middle fossa cyst causing mass effect, intracranial hypertension, and neurological deterioration > 2 years after temporal lobectomy for complex partial seizures. An enlarging cyst with cerebrospinal fluid imaging characteristics was detected on neuroimages 1 year before neurological deterioration. The patient was treated by the insertion of a cystoperitoneal shunt for cerebrospinal fluid diversion. The potential causes of an acquired arachnoid cyst and hydrocephalus are discussed. PMID- 8727835 TI - Subdural hemorrhage caused by de novo aneurysm complicating extracranial intracranial bypass surgery: case report. AB - A case of a de novo aneurysm leading to subdural hemorrhage after right extracranial-intracranial bypass surgery is described. After an uneventful 2 month postoperative course, the patient experienced sudden onset of occipital headache with vomiting. Radiological study disclosed an acute subdural hematoma in the right temporo-occipital region and a newly formed aneurysm at the site of patent superficial temporal artery-middle cerebral artery anastomosis. The anastomotic portion with the ruptured aneurysm was resected en bloc after alternative occipital artery-middle cerebral artery bypass, and the cut end of the superficial temporal artery was successfully used for end-to-side reanastomosis to the other middle cerebral artery branch. The histological examination of the ruptured aneurysm revealed the features of a true aneurysm. PMID- 8727836 TI - Intra-aortic balloon pump counterpulsation in the management of concomitant cerebral vasospasm and cardiac failure after subarachnoid hemorrhage: technical case report. AB - We report two patients who had symptomatic cerebral vasospasm and cardiac failure after aneurysmal subarachnoid hemorrhage and who were treated successfully with intra-aortic balloon pump counterpulsation therapy. Both patients developed congestive heart failure and pulmonary edema while receiving postoperative hypertensive, hypervolemic, hemodilutional (Triple-H) therapy for symptomatic cerebral vasospasm. Both cases of cardiac failure were refractory to maximum pressor and inotropic infusions. Intra-aortic balloon pump counterpulsation was used to optimize cardiac performance to allow continuation of Triple-H therapy and to maintain adequate cerebral perfusion in an attempt to decrease the risk of cerebral ischemic complications. Both patients have had good long-term outcomes. These two cases illustrate the potential usefulness of the intra-aortic balloon pump as an adjunct to Triple-H therapy in patients with symptomatic cerebral vasospasm and cardiac failure. To our knowledge, this report documents the first clinical application of this adjunctive therapy for vasospasm after aneurysmal subarachnoid hemorrhage. PMID- 8727837 TI - Mutism and pseudobulbar symptoms after resection of posterior fossa tumors in children: incidence and pathophysiology and transient cerebellar mutism after posterior fossa surgery in children. PMID- 8727838 TI - Simple risk predictions for arteriovenous malformation hemorrhage. PMID- 8727840 TI - Perception of the impact of a child's chronic illness: does it predict maternal mental health? AB - Longitudinal data were used to investigate direct and indirect effects of demographic factors, parameters of chronic health conditions, service use variables, and perception of family impact of illness on mental health of 169 mothers of children with diverse chronic health conditions. Bivariate analyses indicated that baseline assessments of demographic factors, condition parameters, and service use were: (1) generally unrelated to maternal mental health at 1 year and (2) modestly related to maternal perception of the condition's impact on the family 1 year later. Perceived impact and maternal mental health themselves were moderately related. Multivariate analyses indicated that the need to watch for changes in the child's condition, the presence of communication or speech problems, and the number of hospitalizations in the previous year predicted maternal perceptions of the impact of the condition 1 year later. Mothers perception of impact had a direct effect on mothers' mental health after accounting for condition parameters. Results suggest that condition parameters and service use can affect maternal mental health through mothers' perceptions of the impact of the condition on family life. Implications for interventions to prevent maternal health problems are discussed. PMID- 8727839 TI - Feeding interactions in infants with very low birth weight and bronchopulmonary dysplasia. AB - Infants with very low birth weight (VLBW) are at increased risk for feeding disorders that can affect growth and development. One hundred and forty one mother-infant pairs were compared [55 with infants with high medical risk due to infant VLBW and bronchopulmonary dysplasia (BPD), 34 VLBW without BPD, and 52 term infants] on operationally defined measures of feeding behaviors and maternal self-report of depression and anxiety. Mothers of VLBW infants with and without BPD spent more time prompting their infants to feed when their infants engaged in nonfeeding behavior. Despite increased maternal efforts, infants with BPD took in less formula, spent less time sucking, and spent a greater proportion of time nonfeeding. VLBW infants without BPD were equivalent to term infants in percentage of time sucking and in volume of formula ingested and were more likely to take in higher calories than infants with BPD. Mothers of VLBW infants with and without BPD were also more likely to report clinically significant symptoms of depression and anxiety than mothers of term infants. Because mothers of VLBW infants who were more depressed or anxious were less likely to verbally prompt their infants to eat, maternal psychological symptoms should be considered in assessing interactions of VLBW mother-infant dyads. PMID- 8727841 TI - Childrearing attitudes among parents of very low birth weight and normal birth weight children. AB - The childrearing attitudes of parents of school-age children born at weights under 1500 g (VLBW) were compared with those of parents of age- and gender matched children born at full term (NBW) to determine whether there were systematic differences between the two groups and whether parental attitudes were associated with child outcomes. Parents completed a self-report measure of childrearing attitudes and provided information on the child's health since birth. The children were given measures of IQ and self-concept, and their teachers rated their social and academic competence. Neonatal morbidity and subsequent need for hospitalization were unrelated to parental attitudes. Parents of VLBW children reported less use of guilt as a control strategy. They were also less child-centered, particularly if their children had chronic respiratory or ear-nose-throat problems. Greater parental warmth, less control through guilt, and less parental detachment were associated with more socially competent behavior and more positive self-concept in 9-year-old VLBW children. PMID- 8727842 TI - A survey of physician use of treatment methods for functional enuresis. AB - We report a survey of treatment methods used by primary care physicians in Worcester, Massachusetts in the treatment of functional enuresis. The results suggest a predominance in the use of psychobehavioral over pharmacologic treatment methods and are in contrast to previous studies. Ninety-one percent of physicians reported recommending the use of rewards to control enuresis. Eighty percent of physicians reported using the "bell-and-pad" conditioning method for treatment of functional nocturnal enuresis, contrasted with 5% of physicians reported in previous studies. Fifty-two percent of physicians reported using medication for treatment of functional nocturnal enuresis, despite substantial evidence of its dubious efficacy. PMID- 8727843 TI - Developmental sophistication among adolescents of negotiation strategies for condom use. AB - Condoms are an important method of reducing the risk of sexually transmitted disease among sexually active adolescents. This study used a developmental approach to examine the negotiation of condom use. The Interpersonal Negotiation Strategy Interview was administered to adolescents using two vignettes that were focused on the negotiation of "safer sex" behaviors. The sample consisted of 196 girls and 92 boys recruited from their primary care clinic or Job Corps. The mean scores for each problem-solving step were at the subjective, unilateral stage, which indicates that problems are resolved through one-way methods. Current educational strategies and the lack of mutuality in adolescents' sexual relationships influence the level of negotiation skills. Interventions need to be at the appropriate developmental level but also need to encourage progress toward collaborative relationships. PMID- 8727844 TI - Training in early cancer detection and anxiety in adolescent males: a preliminary report. AB - Because of concerns about increasing anxiety in a population that is at relatively low risk for developing cancer, some investigators have recommended that training in testicular self-examination (TSE) not be provided to adolescent males. However, empirical findings documenting a link between training in cancer detection and increased anxiety in adolescent males are lacking. In this preliminary study, we assessed whether training in TSE was actually associated with elevated state anxiety in two samples of adolescent males (29 ninth graders and 30 college underclassmen). For both groups, anxiety scores were well within normal limits at postassessment, indicating that a purported cost of the procedure may in fact not exist. PMID- 8727845 TI - Maternal education and maternal responsiveness as predictors of play competence in low birth weight, premature infants: a preliminary report. AB - Videotaped play interactions of 182 low birth weight, premature toddlers and their mothers were used to investigate whether a model including maternal responsiveness together with level of maternal education was a better predictor of play maturity scores of the children than a model consisting of education level alone. The videotapes were coded using the responsiveness subscale of the Maternal Behavior Rating Scale. Regression analyses revealed that maternal education alone significantly predicted play scores, accounting for 9% of the variance. However, the model that included both maternal responsiveness and maternal education together also significantly predicted play scores and accounted for 20% of the variance. Including responsiveness together with maternal education significantly increased the proportion of variance accounted for by maternal education alone. Implications of these findings are discussed. PMID- 8727846 TI - Parents as teachers: an integral component of a developmental and behavioral pediatrics curriculum. PMID- 8727847 TI - Crack cocaine exposure in preschool children: focusing on the relevant issues. PMID- 8727848 TI - School underachievement in the fifth grade. PMID- 8727849 TI - Biological correlates of learning and attention: what is relevant to learning disability and attention-deficit hyperactivity disorder? AB - Advances in the related fields of cognitive neuroscience and neuropsychology offer intriguing reformulations of the clinical entities of attention-deficit hyperactivity disorder (ADHD) and learning disabilities (LD). The definition of such terms as attention and learning used in cognitive neuroscience differ from their use in clinical practice and emphasize such processes as intention, working memory, and executive function. Research suggests that neither attention nor long term memory are the critical cognitive correlates of ADHD or LD. Rather, encoding processes, particularly working memory, are identified as deficiencies for children with these conditions. Furthermore, intention and inhibition appear to be particularly impaired in children with ADHD, who exhibit broader deficits in so-called executive function. Findings from cognitive neuroscience offer explanations of the neuroanatomical and neurophysiological underpinnings of learning problems and the frequent comorbidity of LD and ADHD. PMID- 8727850 TI - The extent and importance of research to developmental and behavioral pediatrics. PMID- 8727851 TI - Pleuroscopy: therapeutic applications. AB - Pleuroscopy is mostly regarded as a diagnostic procedure. Although therapeutic uses of pleuroscopy were popular when pulmonary tuberculosis was common, they are less well known today. This review of modern therapeutic pleuroscopy is based on both personal experience and previous reports. We have grouped the purposes of therapeutic pleuroscopy as 1) to provoke formation of pleural adhesions in the management of pleural effusion, recurrent pneumothorax, chylothorax or (in selected cases) empyema, 2) to divide adhesions in persistent pneumothorax, 3) to perform pleural toilet in the fibrinopurulent stage of empyema, 4) to retrieve foreign bodies, and 5) to achieve haemostasis and removal of clotted blood following operation or trauma. These applications of pleuroscopy should be studied and popularized so that the method can attain recognition as a revived therapeutic procedure. PMID- 8727852 TI - In vivo assessment of luminal cross-sectional areas and circumferential tension strain relations of the porcine aorta. AB - The objectives were to measure the pressure--cross-sectional area relations and intrinsic stiffness of the porcine aorta in vivo. METHODS: Measurements were made in 12 pigs, weight 30, 50 or 75 kg, proximal and distal to the bifurcation of the renal arteries, using an electrical impedance system which was located inside a balloon mounted on a catheter. Vessel cross-sectional area (CSA) was assessed by measuring impedance of the fluid inside the balloon during distension. RESULTS: In vitro testing demonstrated the accuracy and reproducibility of impedance planimetry. In vivo steady-state CSA values showed non-linear relationship with rising distension pressures. Mean CSA values rose with increasing weight of the pigs. the suprarenal aorta was larger than the infrarenal only in 75 kg pigs (p < 0.05). At three measurements the within-subject variation was 0.89% and the between-subject variation 99.11%. Nitroglycerine infusion produced only minor, insignificant CSA increase, indicating negligible tone in the abdominal aorta. All segments showed circumferential wall strain-tension relations with exponential behaviour uninfluenced by weight class or site in the aorta. CONCLUSION: Information of vascular mechanics in vivo is obtainable by catheterization of the abdominal aorta. PMID- 8727853 TI - Pulmonary artery abnormalities in tetralogy of Fallot and relation to late physical performance. AB - Preoperative angiograms were reviewed in 86 (52%) of 165 patients who underwent intracardiac repair of tetralogy of Fallot in 1966-1976. The median age at repair was 7 years (range 9 months-55 years). Hypoplasia, stenosis or atresia of the pulmonary and aortopulmonary collateral arteries were present in 58 patients (67%). Hypoplasia of the main pulmonary artery was associated with hypoplasia of the pulmonary artery branches. In patients who received a transannular patch the pulmonary arteries were smaller than in those without patch. Three of five pulmonary artery branch stenoses were located at the site of a previous palliative shunt. The postrepair right ventricular/left ventricular systolic pressure ratio (PRV/LV) measured in the operating room was compared with a predicted ratio based on pulmonary artery dimensions. High PRV/LV was associated with poor prognosis. In survivors the postrepair PRV/LV had fallen significantly 13-26 years later. Work capacity and pulmonary function at follow-up were not influenced by presence of pulmonary artery abnormalities. We conclude that such abnormalities, although common in patients with tetralogy of Fallot, did not per se influence physical performance in long-term survivors. PMID- 8727854 TI - Oesophageal squamous cell carcinoma after gastrectomy for benign ulcer disease. AB - Of 684 patients treated for squamous cell carcinoma of the oesophagus in 1982 1993, 19 (2.8%) had previously undergone partial gastrectomy for benign ulcer disease. The average interval between gastrectomy and diagnosis of oesophageal carcinoma was 14 years. In six of the 19 (31.6%) cases the carcinoma was in the lower thoracic oesophagus, an incidence not significantly higher than the 25% in the total series. The oesophageal resectability rates were 52.6% in the gastrectomized patients and 44.4% in the total series (intergroup difference not significant). Of the ten gastrectomized patients who underwent oesophageal resection, only one had oesophagitis at the oesophagogastric junction, without significant histologic characteristics. The relationship between oesophageal squamous cell carcinoma and previous gastrectomy for benign ulcer disease may be incidental. PMID- 8727855 TI - Relationships between divisions of the lingular bronchus and vascularization patterns in the lingula. AB - In 100 left human lungs the main bronchus, the pulmonary artery and the pulmonary veins were injected with 65% methyl methacrylate (Duracryl) and then digested in sulphuric acid. The resulting specimens were studied concerning the divisions of the lingular bronchus and the types of arterial and venous vascularization of the lingula. As a rule the lingular bronchus divided into two segmental bronchi. A single lingular artery was found in 80% of the cases and a single lingular vein in 58%. Atypical bronchial divisions were almost always associated with unusual types of vascularization. Patterns of bronchial division showed complete concordance with those of arterial vascularization of the lingula in 64% of the lungs and consistency with venous drainage patterns in 54%. PMID- 8727856 TI - A new no-touch preparation technique. Technical notes. PMID- 8727857 TI - Delayed intrathoracic herniation of the stomach with pleural empyema due to diaphragmatic stab wound. AB - A 32-year-old man received a left-sided thoracic stab wound, which was primarily treated with percutaneous tube thoracostomy. Ipsilateral empyema appeared 8 weeks later and subsequent investigations revealed herniation of the stomach through the diaphragm. The diaphragmatic rupture and a perforation in the gastric wall were repaired at thoracotomy. The literature on such wounds is reviewed. PMID- 8727858 TI - Effects of ovariectomy on mechanical properties and collagen content in rabbit lower urinary tract smooth muscle. AB - Fourteen female rabbits underwent ovariectomy or were sham-operated six weeks before investigation. Detrusor and urethral strips (longitudinal and circular urethral muscle fibers were studied separately) were prepared and length-tension relations studied in organ baths experiments. In addition, the specimens were examined for collagen content. Maximal active tension in the detrusor, longitudinal and circular urethral preparations, determined as the response to K+ (124mM), was reached when the length of the strips was 178 +/- 8% (n = 7), 153 +/ 9% (n = 5), and 127 +/- 5% (n = 7) of the resting length, respectively. Ovariectomy did not alter the length for development of maximal active tension. In detrusor strips, ovariectomy caused a significant increase in maximal active tension from 39 +/- 7 mN (sham-operated rabbits) to 79 +/- 11 mN (p < 0.01), despite an unchanged relative amount of smooth muscle. The maximal active tension produced in the urethra was lower (15-25 mN) than in the detrusor, and not significantly affected by ovariectomy. Approximately 40% of the dry defatted tissue weight of the detrusor consisted of collagen, whereas corresponding value in the urethra was 50-60%. Ovariectomy had no effect on tissue collagen concentrations. In conclusion, ovariectomy for six weeks did not affect the passive mechanical properties or the collagen concentration in rabbit lower urinary tract smooth muscle, but increased the responsiveness of the detrusor muscle to K+ (124 mM). PMID- 8727859 TI - Fertility after autotransposition of vas: a comparative study of absorbable and nonabsorbable suture material in rats. AB - The anastomosis patency after autotransposition of vas (ATV) using an absorbable and a nonabsorbable suture material (10-0 Vicryl vs Ethilon) and the fertility have been investigated in a rat model. The patency rate and fertility in the absorbable group were 84% and 48%, respectively, compared with 93% and 59% in the nonabsorbable group. Sperm granulomas occurred in 60% and 48%, respectively. Sperm antibodies were present in 36% of the cases but did not have an impact on the fertility. The fertility after ATV seems to be comparable to the fertility in experimental and clinical vasovasostomy series. Further, the two suture materials appear equivalent with respect to patency and fertility in ATV. PMID- 8727860 TI - Bacterial adhesion and biofilm formation on various double-J stents in vivo and in vitro. AB - The aim of this study was to determine whether bacterial adhesion and biofilm formation occur on various ureteric stent materials in vivo and in vitro and whether there is any correlation between biofilm formation and urinary tract infection. Using an open surgical procedure, 23 double-J stents of 8 different types were inserted in the ureters of 13 piglets. After 6 weeks intubation, 2 (9%) of the 23 stented renal units yielded positive urine culture. Scanning Electron Microscopy (SEM) showed sporadic bacteria on 7 (30%) of the 23 indwelling ureteric stents and bacterial biofilms on only 2 (9%), those indwelling the renal units which yielded positive urine cultures. Precut segments of the 8 double-J stent brands were incubated with 5 different bacterial strains. Microbiological and SEM analysis of the stents revealed that only 0.1-0.01% of the bacterial inoculum adhered to the stents. There was no statistically significant difference in bacterial adherence to the various stent materials. Interestingly, the two Escherichia coli strains with P-fimbriae adhered significantly more than the two without P-fimbriae. Sporadic bacterial adhesion to ureteric stents seemed to have little clinical significance whereas biofilm formation seemed to result in urinary tract infection. None of the tested stents proved to be superior to the others in this respect, and adhesion appeared to depend on the virulence of the bacteria rather than the properties of the biomaterials. PMID- 8727861 TI - Can dipstick screening for hematuria identify individuals with structural renal abnormalities? A sonographic evaluation. AB - The present study presents data from a population study of 659 randomly selected adult volunteers. All participants had a renal sonographic examination and at the same time had their urine tested for hematuria by dipstick. Thirty participants (5%) had dipstick hematuria. Similar rats of abnormal renal findings were obtained in the hematuria-positive and hematuria-negative groups, 10% and 8%, respectively. Our results suggest that dipstick investigation for hematuria in mass-screening programs is not effective for the detection of structural renal abnormalities. PMID- 8727862 TI - Quantitative bone scintigraphy in patients with prostatic carcinoma treated with LH-RH analogues. AB - In 14 men with prostatic carcinoma, quantitative scintigraphy of the vertebrae from Th10 to L5 was performed before and 2 weeks and 2 and 6 months after start of treatment with luteinizing hormone-releasing hormone (LH-RH) analogues. Serum prostate-specific antigen (PSA) was also determined. The patients with normal bone scintigram showed no change in gamma camera count rate during the study, but fall in PSA values. The patients with abnormal bone scintigram responded to treatment with flare phenomenon, with increased count rate at 2 weeks, followed by fall to pretreatment level at 2 months. PSA showed decrease as early as 2 weeks after the start of treatment. Bone scintigraphy was found to be useful before therapy, especially in patients with elevated PSA levels, and after 2 months, when the flare phenomenon had subsided. Serial measurement of PSA provided a guide to disease activity. PMID- 8727863 TI - Prospective randomized open study between ciprofloxacin and a combination of sulfadiazine and trimethoprim in antibiotic prophylaxis in connection with transurethral prostatectomy. AB - A randomized open prospective study with antibodies was conducted on 398 cases of TURP to compare the efficacy of either 250 mg of ciprofloxacin twice daily (CF) or 500 mg/160mg of sulfadiazine-trimethroprim twice daily (ST) in the prevention of urinary and other infection complications. The medication was started on the evening preceding the operation and continued up to the day following the removal of an indwelling catheter. The preoperative risk factors were similar in both groups. The groups did not differ in terms of the duration of the operation, the volume of irrigation fluid or the weight of the resected chips. The incidence of immediate complications was 26% in the CF group and 20% in the ST group. All the complications were minor. There were three serious late complications, one intertervertebral discitis and one coxitis in the CF group and one endocarditis in the ST group. The patient with endocarditis died. One the removal of the indwelling catheter on the third postoperative day, bacteriuria was detected in 3% of the patients in the CF group and in 9% in the ST group. The difference was statistically significant (p < 0.05). One month after TURP, bacteriuria was detected in 7% in both study groups. It is concluded that ciprofloxacin may be more effective to preventing immediate urinary tract infections after TURP than a combination of sulfadiazine and trimethoprim. There were three serious late complications. As late as one month after TURP, bacteriuria was still diagnosed in about 7% of all patients, which is post-TURP monitoring of urinary values is important. PMID- 8727864 TI - An estimate of the life-time cost of surgical treatment of patients with benign prostatic hyperplasia in Sweden. AB - Two clinical series (n = 96 + 90) and one record-linkage study (n = 492) were used for estimation of the health care utilization for the treatment of BPH patients, mainly by TURP, in Sweden during one year before and 5-7 years after surgery. The total cost for a single patient amounted to ca. 33000 SEK in 1900 prices (5850 USD). Costs for surgery dominated and for a TURP amounted to about 70% of the Total. The costs during one year preoperatively and 5 years postoperatively each amounted to 15% of the total costs. In the present study the outcome of surgery was similar to other reports from the same period. The surgical mortality was 0.4% and the readmission rate because of complications of surgery or manifestations of BPH was 25% after 7 years of observation. Of the patients 11% were reoperated on within 7 years. When transurethral resection of the prostate (TURP) replaced open surgery in Sweden during the 1970's it had several of the attributes of the new methods introduced for treatment of benign prostate hyperplasia (BPH) currently in use. However, the spread of TURP resulted in wider indications for surgery and an increase in the total number of surgical procedures. An important argument for the adoption of the new, less invasive methods for treatment of BPH is the lower cost. To make a fair comparison of the costs of different methods for treatment of BPH the long-term costs have to be included in the calculation. PMID- 8727865 TI - The symptomatic outcome of transurethral prostatectomy evaluated with the DAN-PSS 1 score system. AB - We assessed the symptomatic outcome of transurethral prostatectomy with the newly developed self-administered questionnaire, Danish Prostate Symptom Score (DAN-PSS 1), in men suffering from uncomplicated benign prostatic hyperplasia (BPH). The questionnaire was filled in adequately by 53 men, median age 69 (range 52-82), preoperatively and 2, 4 and 6 months after transurethral prostatectomy (TUR-P). The DAN-PSS-1 symptom score showed a high degree of sensitivity as the preoperative total score was median 20 (quartiles 14-36) and decreased to 4, 0 and 1 (quartiles from 0-4) at the 2, 4, and 6-month control respectively. The symptoms related to voiding were relieved totally after 2 months while symptoms related to storage were first relieved completely at 4-month control. Bother from symptoms was relieved after 2 months, although some symptoms still remained. The questionnaire was internally consistent with a reliability coefficient, Cronbach's alpha (alphaCr), of 0.73. We conclude that the DAN-PSS-1 questionnaire is an efficient tool among others in the indication and evaluation of treatment of uncomplicated BPH. PMID- 8727866 TI - Stabilization of the nipple base with titanium rings when reoperating patients with continent urostomies for nipple dysfunction. AB - Nipple dysfunction in Kock reservoirs used for continent urinary diversion is usually caused by a flattening out of the intestinal nipple due to a lateral tore at the base of the nipple when the reservoir expands. This study describes a new method to repair dysfunctioning nipples based on the use of titanium rings applied outside the reservoir around the base of the nipple. Experiments on dogs and rats have shown that rings and plates of titanium rapidly attach to the serosal surface of ileum exposed to urine with only minor signs of inflammation. Ten patients with Kock reservoirs for urinary diversion have been reoperated for nipple dysfunction due to nipple sliding with elongation of the nipple and fixation of the nipple base with titanium rings. The functional result is excellent in 9 out of the 10 patients. No signs of erosion of the rings into the reservoirs have been noted. With this new method for nipple repair the need for using more intestine to construct a new nipple is eliminated. PMID- 8727867 TI - Enuresis nocturna can be provoked in normal healthy children by increasing the nocturnal urine output. AB - In a large proportion of enuretics, the enuresis episodes are accompanied by high urine output at night. At this study we provoked a high urine output in normal non-enuretic children in order to investigate whether volume provocation per se could cause enuresis. In six of ten children it was possible to provoke nine enuresis-like episodes. The enuresis volumes were small with no relation to functional bladder capacity. It is concluded, that increased urine output at night may produce enuresis-like episodes even in normal children. PMID- 8727868 TI - Does prophylactic treatment with felodipine, a calcium antagonist, prevent low osmolar contrast-induced renal dysfunction in hydrated diabetic and nondiabetic patients with normal or moderately reduced renal function? AB - Twenty-seven patients (15 diabetics and 12 non-diabetics) with normal to moderately reduced renal function underwent femoral angiography with a low osmolar contrast agent, iohexol (Omnipaque), under perexaminatory hydration. Fourteen patients were randomised to pretreatment with oral felodipine extended release (Plendil) 10 mg and 13 patients to placebo 3-4 h before angiography. GFR measured with [51Cr] EDTA-clearance decreased 24 hours after the angiography in the felodipine group from GFR 52.5 +/- 18.6 (mean +/- SD) to 46.2 +/- 16.5 ml/min (p < 0.01) and in the placebo group from 70.6 +/- 18.6 to 62.6 +/- 26.4 ml/min (p < 0.01). Serum creatinine increased significantly in the felodipine group from 128 +/- 61 to 139 +/- 67 mumol/l (p < 0.05) but not in the placebo group (122 +/- 54 to 125 +/- 51 mumol/l (ns)). The values of serum creatinine returned to baseline levels 7 days after angiography. During hydration there was only a slight reduction of GFR after angiography with iohexol. Thus, felodipine had no major effect on GFR after iohexol but, as baseline GFR tended to be lower in the felodipine pre-treated patients, it might have had some renoprotective effect in patients with more advanced renal failure. PMID- 8727869 TI - Acute blockage of the renin system and differential renal vein renin determinations in the diagnosis of renovascular hypertension. AB - For validation of differential renal vein renin determinations in the diagnosis of renovascular hypertension (RVH), we investigated 102 patients suspected of suffering from RVH before and 1 h after administration of 25 mg captopril. Sensitivity, specificity and posterior probability for renin ratio (RR) and renin secretion (RS) were calculated based on 44 patients with proven RVH and 58 patients with primary hypertension (PH) using discriminant analysis. There is good (> 95%) and identical specificity of both variables under all conditions, whereas sensitivity remains poor even after Captopril administration (RR 23% vs. 32%; RS 20% vs. 34%). The posterior probabilities obtained by discriminant analysis revealed a cut-off point of 2.5 for the renin ratio and of 1.9 for the renin secretion. No change is observed after ACE inhibition. We conclude that the acute blockade of the renin system by captopril in differential renin sampling yields no advantages in diagnosing RVH and that there is no difference between RR and RS in the diagnosis of RVH. PMID- 8727870 TI - True hermaphroditism. AB - True hermaphroditism was found in a phenotypically normal boy admitted to the urology department with the diagnosis of right undescended testis. The tissue expected to be cryptorchid proved to constitute an ovary, uterus and salpinx. Normal left testicular tissue was found at biopsy. The patient's genotype was 46 XX. PMID- 8727871 TI - Dicentric Y chromosome without evidence of mosaicism in an azoospermic male. AB - A phenotypically male of dicentric Y chromosome with no evidence of sex chromosomal mosaicism is reported. Staining of fluorescence in situ hybridization using Y chromosome centromere-specific DNA probe showed the presence of two bright fluorescent spots in Y chromosome. Testicular histology revealed maturation arrest at the stage of primary spermatocyte. It is suggested that structural anomaly of the Y chromosome is related to the disturbance of spermatogenesis. PMID- 8727872 TI - Gangrene of the penis due to strangulation by a metallic ring. AB - We present a case of a 32-year-old crippled patient who presented with gangrene of the penis due to strangulation by a magnetic metallic ring which is used to increase potency and prolong erection. The ring was removed and conservative treatment was tried for eight days but failed. Amputation of the penis was done was refashioning of the urethra at the tip of the stump. PMID- 8727873 TI - Evaluation of tuberculosis transmission in a community by 1 year of systematic typing of Mycobacterium tuberculosis clinical isolates. AB - Interhuman transmission of Mycobacterium tuberculosis was investigated by using molecular typing, including restriction fragment length polymorphism with probes IS6110, DR (direct repeat) and PGRS (polymorphic GC-rich sequence) and a PCR method using the inverted repeat sequences of IS6110 as primers. From 105 patients hospitalized for tuberculosis during a 1-year survey in three hospitals in Paris, France, 111 isolates were collected and analyzed. Eighty-eight patients were infected with genetically different isolates, demonstrating the clonal heterogeneity of M. tuberculosis in these patients originating from various geographical areas. Fourteen patients were infected by strains clustered with identical fingerprints. An epidemiological relatedness was demonstrated for isolates from only seven of these patients. Thus, the typing of isolates from all tuberculous patients in hospitals during 1 year allows the detection of transmission in the general community. This would improve the case findings, thereby further improving the detection of outbreaks. PMID- 8727874 TI - Serological and genomic characterization of porcine rotaviruses in Thailand: detection of a G10 porcine rotavirus. AB - A total of 557 fecal specimens collected from piglets with diarrhea in Thailand were examined for rotavirus RNA by polyacrylamide gel electrophoresis. Twenty three, one, and two samples were positive for group A, group B, and group C rotaviruses, respectively. Two samples exhibited two segments found in picobirnavirus RNA. RNA electropherotyping of 23 group A rotaviruses showed that they were classified into five patterns. By serotyping by enzyme-linked immunosorbent assay and PCR, viruses in 3 and 14 specimens were found to be serotype G3 and serotype G10, respectively. For one specimen, containing a serotype G10 virus (strain P343), virus was isolated in MA-104 cells, and the nucleotide sequences of the VP7 and VP4 genes were determined. Comparative sequence analysis and cross-neutralization tests showed that strain P343 has B223 like G10 and UK-like P7 serotype (or VP4 genotype 5) specificities. Rotaviruses having such antigenic specificities have not been detected in piglets. Thus, the interspecies transmission of rotaviruses between cows and pigs was suggested. PMID- 8727876 TI - Evaluation of Amplicor MTB test as adjunct to smears and culture for direct detection of Mycobacterium tuberculosis in the French Caribbean. AB - A total of 784 specimens collected from 370 individuals between January and August 1995 were analyzed by using the Amplicor Mycobacterium tuberculosis test (Roche Diagnostic System, Basel, Switzerland), a PCR-based test for the direct detection of organisms of the M. tuberculosis complex. The PCR results were compared with standard bacteriological data, including those obtained by acid fast microscopy, culture, and biochemical identification as well as final clinical diagnosis for each patient. Several parallel controls were used: the kit DNA positive control, 10(3) CFU of M. tuberculosis, and three negative controls for each independent assay. No false-positive PCR results were obtained, and overall, M. tuberculosis was detected in 20 of 370 individuals screened. Five additional patients during the same time were found to be infected with mycobacteria other than tubercle bacilli; their specimens gave positive smear and/or culture test results, but Amplicor tests were always negative. The sensitivity, specificity, positive predictive value, and negative predictive value for the Amplicor MTB test compared with culture per specimen were 76.7, 97.7, 66.0, and 98.6%, respectively. For resolved cases, these values were, respectively, 69.4, 100, 100, and 96.8%; however, the sensitivity and negative predictive value increased to 90.9 and 99.2%, respectively, if PCR-negative nonrespiratory specimens (gastric washings) were not considered. When only specimens from proven tuberculosis patients were considered (n = 114) and the sum of PCR-positive and/or culture-positive samples from proven tuberculosis patients was considered the total number of positive samples, PCR had a sensitivity of 83.3% compared with 71.6% for culture. Results per patient (about three samples each) yielded 100% sensitivity and 100% specificity. We conclude that the Amplicor MTB test is highly specific and rapid for routine use in a clinical laboratory. However, in order to obtain a higher degree of sensitivity, it should be run as an adjunct to smears and culture with at least three samples for each patient, and a single-sample PCR-negative results must be considered carefully because of potential false-negatives. PMID- 8727875 TI - Comparative evaluation of NASBA HIV-1 RNA QT, AMPLICOR-HIV monitor, and QUANTIPLEX HIV RNA assay, three methods for quantification of human immunodeficiency virus type 1 RNA in plasma. AB - Three commercial assays for quantifying plasma human immunodeficiency virus type 1 (HIV-1) RNA were evaluated. The assays differed in their sample volumes, the means of preparing samples, and methods of amplification and detection. Plasma samples were obtained from 36 HIV-1-infected patients representing all stages of HIV-1 infection and were analyzed as coded specimens. Measurement of HIV-1 RNA baseline levels revealed no significant difference in sensitivity between the three assays. The assays were also applied to the quantitation of HIV-1 RNA levels in the plasma of patients who were changing their antiretroviral therapy. The changes measured in HIV-1 RNA levels in plasma in response to therapy were comparable by the three assays. No close correlation was found between the amount of HIV-1 RNA and the CD4 T-cell count; HIV-1 RNA assays were more sensitive than p24 antigen assays as an indicator of plasma viremia. Overall, the study demonstrates that all three quantitative assays for HIV-1 RNA can be used to measure the HIV-1 RNA copy number representing the HIV-1 viremia status in patients with HIV-1 infection. Since this copy number is likely to be useful in monitoring the effectiveness of antiviral therapy, these quantitative assays for HIV-1 RNA are ready to be built into clinical trials. PMID- 8727877 TI - Ability of RapID Yeast Plus System to identify 304 clinically significant yeasts within 5 hours. AB - The RapID Yeast Plus System (Innovative Diagnostic Systems, Norcross, Ga.) is a qualitative micromethod that uses conventional and chromogenic substrates for the identification of medically important yeasts. The ability of the RapID Yeast Plus system to accurately identify 304 clinical yeast isolates within 5 h was evaluated. The RapID Yeast Plus method correctly identified 286 (94.1%) of strains to the species level without the need for additional tests. A further 12 strains (3.9%) were classified as correct to the genus level or to a low probability identification with two or more possibilities. In these latter cases, additional tests were required to delineate the correct identification. Organisms in the latter group comprised Candida parapsilosis (n = 1), Candida tropicalis (n = 1), Candida ciferrii (n = 1), Candida guilliermondii (n = 2), Candida humicola (n = 1), Candida kefyr (n = 1), Cryptococcus neoformans (n = 1), and Rhodotorula rubra (n = 4). Six strains (2.0%) were misidentified or did not yield codes in the manufacturer's database. These included one Candida utilis (identified as Candida famata/Candida guilliermondii), one Trichosporon beigelii (identified as Cryptococcus neoformans), one Candida diddensiae (identified as Candida albicans), one Candida membranaefaciens (identified as Candida parapsilosis), one Candida norvegensis (identified as Candida zeylanoides), and one Candida catenulata (no code) isolate; the last four strains are not included in the firm's current database. The RapID Yeast Plus system yielded excellent results and may be recommended for use in the routine laboratory for accurate same-day identification of clinically significant yeasts. PMID- 8727879 TI - Ribotyping: a tool for taxonomy and identification of the Nocardia asteroides complex species. AB - Ribotyping has been evaluated as a taxonomic tool for studying bacteria belonging to the Nocardia asteroides complex. The probe used was obtained by PCR from DNA extracted from the type strain Nocardia asteroides ATCC 19247, a sequence that codes partly for 16S rRNA. Interpretation of hybridization after EcoRI restriction of total DNA of 21 strains of the N. asteroides complex allowed for the identification of only four different ribotypes: two related to N. asteroides sensu stricto, one related to N. farcinica and one related to N. nova. These results confirm the classification established by Tsukamura in 1982 (M. Tsukamura, Microbiol. Immunol. 26:1101-1119, 1982) as well as the heterogeneity of N. asteroides sensu stricto species. Ribotyping may also provide a major identification technique for the species belonging to the N. asteroides complex. PMID- 8727878 TI - Three species of Borrelia burgdorferi sensu lato (B. burgdorferi sensu stricto, B afzelii, and B. garinii) identified from cerebrospinal fluid isolates by pulsed field gel electrophoresis and PCR. AB - A total of 36 European Borrelia burgdorferi sensu lato cerebrospinal fluid isolates (mainly from southern Germany) were analyzed by pulsed-field gel electrophoresis (PFGE) for large restriction fragment pattern (LRFP) and linear plasmid profiles. Analyzing this large panel of isolates, we detected all three species of B. burgdorferi sensu lato pathogenic for humans in cerebrospinal fluid from patients with Lyme neuroborreliosis by PFGE typing after MluI digestion: 21 B. garinii (58%), 10 B. afzelii (28%), and 4 B. burgdorferi sensu stricto (11%) strains as well as 1 isolate with bands characteristic of both B. afzelii and B. garinii. Species classification by PFGE typing was confirmed by 16S rRNA-specific PCR. Eighteen isolates (11 B. garinii, 6 B. afzelii, and 1 B. burgdorferi sensu stricto isolate) were further characterized by LRFP with four different restriction enzymes (ApaI, KspI, SmaI, and XhoI). All B. afzelii isolates showed identical patterns for each restriction enzyme group. Considerable heterogeneity was demonstrated within the B. garinii group. Subsequent analysis of plasmid profiles revealed only marginal differences for B. afzelii strains but different patterns for B. garinii isolates. In one B. afzelii strain we found a linear plasmid of about 110 kbp not described before. LRFP analysis by PFGE is a suitable tool for the molecular characterization of B. burgdorferi sensu lato strains and allows determination not only of the species but also of the subtypes within B. garinii. PMID- 8727880 TI - Clinical evaluation of the Roche AMPLICOR PCR Mycobacterium tuberculosis test for detection of M. tuberculosis in respiratory specimens. AB - The reliability of the Roche AMPLICOR Mycobacterium tuberculosis test (AMPLICOR MTB) for the diagnosis of pulmonary tuberculosis was evaluated by testing 956 respiratory specimens from 502 patients and comparing results with results by culture and medical history. Of those 135 specimens that were culture positive for mycobacteria, 61 specimens from 31 patients grew M. tuberculosis. Fifty-two specimens were smear positive for acid-fast bacteria (AFB); M. tuberculosis was isolated from 41 of these specimens. On initial testing, the sensitivity and specificity of the AMPLICOR MTB assay, compared with culture, were 78.7 and 99.3%, respectively. After resolution of discrepancies (by review of medical history), the sensitivity, specificity, and positive and negative predictive values of the AMPLICOR MTB assay were 79.4, 99.6, 92.6, and 98.6%, respectively. Two specimens from two patients with no clinical evidence of tuberculosis were AMPLICOR MTB positive and culture positive for Mycobacterium avium complex. For AFB smear-positive specimens, the sensitivity, specificity, and positive and negative predictive values of AMPLICOR MTB were 97.6, 100, 100, and 90.9%, respectively. For AFB smear-negative specimens, the sensitivity, specificity, and positive and negative predictive values of AMPLICOR MTB were 40.0, 99.5, 69.2, and 98.7%, respectively. Our results support the use of AMPLICOR MTB for rapid diagnosis of tuberculosis in patients whose respiratory specimens are AFB smear positive. Further studies are needed to determine the most clinically relevant and cost-effective use of this assay with AFB smear-negative specimens. PMID- 8727881 TI - Ribotype diversity of Listeria monocytogenes strains associated with outbreaks of listeriosis in ruminants. AB - Ribotyping is a molecular method for the characterization, identification, and typing of bacterial isolates that has value in epidemiological studies. To demonstrate the utility of this technique for typing of Listeria monocytogenes, four outbreaks of epizootic listeriosis in ruminants were investigated through coordinated detection and characterization methods utilizing classical microbiology and nucleic acid-based techniques. L. monocytogenes strains isolated from clinical samples and the silage consumed by the affected animals were ribotyped to establish the causal relationship between feed and the disease outbreak. For all but one outbreak, we were able to isolate L. monocytogenes strains represented by the same ribotype from both clinical and silage samples. Additional L. monocytogenes strains with ribotypes different from those of the respective clinical samples were isolated from all silage samples. This indicates that a diverse population of L. monocytogenes strains exists in farm environments, of which some may be more likely than others to cause disease. PMID- 8727882 TI - Studies of in vivo distribution of bovine herpesvirus type 4 in the natural host. AB - The in vivo distribution of bovine herpesvirus type 4 (BHV-4) was examined by testing nasal and conjunctival exudates, peripheral blood leukocytes, and various organs of experimentally infected calves. For virus detection, a nested PCR assay, virus isolation, and immunohistochemistry were applied. The nervous system and the muscles were free of viral DNA. Liver and intestinal lymph nodes contained low amounts of virus (less than two copies per 1 microgram of cellular DNA). Intestinal, tonsil, thymus, and kidney tissues contained more viral DNA copies (5 to 50 copies per 1 microgram of cellular DNA). The highest amounts of BHV-4 DNA (50 to 500 copies per 1 microgram of cellular DNA) were found in the spleen, lungs, trachea, and nasal epithelium. Amplification of DNA from blood lymphocytes through postinoculation (p.i.) day 48 proved that the virus started to replicate in these cells immediately after inoculation of the calves and that intensive virus growth took place during the 7 to 8 weeks of the infection. The number of virus-infected lymphocytes reached the maximum on p.i. days 22 to 26 and slowly declined thereafter. Virus-infected cells were found only in the spleen on p.i. day 48 by immunohistochemistry. Western blotting (immunoblotting) detected signs of an immune response against 9 of the 29 BHV-4 proteins. PMID- 8727883 TI - Random amplified polymorphic DNA typing versus pulsed-field gel electrophoresis for epidemiological typing of vancomycin-resistant enterococci. AB - Sixty vancomycin-resistant vanA mutant Enterococcus faecium (VRE) isolates, collected during a 40-month period from 48 patients hospitalized in a French Cancer Referral Center, were typed by using random amplified polymorphic DNA (RAPD), and the results were compared with those previously obtained by typing with SmaI pulsed-field gel electrophoresis (PFGE), which is currently recognized as the "gold standard." The discriminating power of RAPD typing, with seven primers and 11 combinations of primers, was tested on 18 strains, and only the most discriminating combination was further tested on the whole collection. We compared the epidemiological usefulness of RAPD typing of 60 clinical VRE isolates with that of SmaI PFGE typing. With primers AP4 and ERIC1R, RAPD generated 30 patterns versus the 36 patterns generated by SmaI PFGE. However, this did not hamper the epidemiologically correct clustering of 15 related strains and the detection of multiple colonization in nine patients. We conclude that this simple RAPD technique is well suited to the epidemiological typing of VRE and the monitoring of its nosocomial spread. PMID- 8727885 TI - Evaluation of mtp40 genomic fragment amplification for specific detection of Mycobacterium tuberculosis in clinical specimens. AB - A PCR assay based on the species-specific mtp40 genomic fragment was developed for the specific detection and identification of Mycobacterium tuberculosis in different uncultured clinical specimens. The aim of the study was to evaluate the clinical applicability of this target DNA in comparison with those of conventional microbiological methods and to compare the results obtained with those obtained after amplification with the IS6110 repetitive element. Discrepant results were interpreted in conjunction with the patients' clinical data, medical histories, and response to therapy. A total of 172 specimens from 162 patients with respiratory symptoms were tested, 101 specimens were obtained from 92 patients clinically suspected of having tuberculosis, and 71 specimens were obtained from 70 patients without known mycobacterial infection. The results of our study suggest that PCR amplification with the mtp40 genomic fragment provides a highly sensitive and specific technique for the detection of M. tuberculosis strains in clinical samples. It allows for the differentiation between M. tuberculosis and other related mycobacteria, including M. bovis, and is more specific than the IS6110 target. For these and other reasons, we propose that the mtp40 assay is a possible alternative for the specific direct detection of M. tuberculosis in clinical laboratories. PMID- 8727884 TI - Isolation and characterization of a unique group of slowly growing mycobacteria: description of Mycobacterium lentiflavum sp. nov. AB - A distinct group of slowly growing mycobacteria was identified on the basis of growth characteristics, biochemical and lipid profiles, and nucleic acid analyses. The isolates showed growth at 22 to 37 degrees C, yellow pigmentation, and negative tests for Tween 80 hydrolysis, nicotinic acid, nitrate reductase, and urease; tests for arylsulfatase, pyrazinamidase, and heat-stable catalase were variable. Analysis of cellular fatty acids by gas-liquid chromatography and mycolic acids by thin-layer chromatography and high-performance liquid chromatography indicated a distinctive pattern which was unlike those of other species. Determination of the 16S rRNA gene sequence showed a unique sequence closely related to Mycobacterium simiae and M. genavense. On the basis of DNA homology studies, we suggest that these organisms are representatives of a novel species, for which the name M. lentiflavum sp. nov. is proposed. PMID- 8727886 TI - Nontoxigenic Vibrio cholerae 01 serotype Inaba biotype El Tor associated with a cluster of cases of cholera in southern India. AB - Thirteen strains of Vibrio cholerae 01 belonging to the Inaba serotype El Tor biotype isolated from patients during an outbreak of cholera in the town of Warangal in southern India were found to be nontoxigenic (NT), since they did not produce cholera toxin or hybridize with DNA probes specific for cholera toxin, Zot, or Ace. The unheated and heated culture supernatants of the NT V. cholerae 01 evoked a rapid cell-rounding effect when introduced on confluent layers of CHO and HeLa cells which could not be inhibited by antiserum against known toxins. Culture supernatants of two representative NT V. cholerae 01 strains caused an increase in short-circuit current in rabbit ileal tissue mounted on an Ussing chamber, and the pattern of increase in short-circuit current was consistent with the presence of a quickly acting toxin like stable toxin. None of the strains of NT V. cholerae 01 hybridized with a DNA probe specific for the heat-stable enterotoxin of V. cholerae non-01, nor did the factor produced by NT V. cholerae 01 resemble the recently described heat-stable enterotoxin produced by enteroaggregative Escherichia coli as determine by a PCR assay. To our knowledge, this is the first report of NT V. cholerae 01 being associated with a cluster of cases of cholera, and it appears that a clone of NT V. cholerae 01 has the potential to cause localized outbreaks of cholera. PMID- 8727887 TI - Usefulness of the secondary probe pTBN12 in DNA fingerprinting of Mycobacterium tuberculosis. AB - A comparison was made between DNA fingerprints of Mycobacterium tuberculosis produced with the insertion sequence IS6110 and those produced with the polymorphic GC-rich repetitive sequence contained in the plasmid pTBN12. A total of 302 M. tuberculosis isolates from the prison system in Madrid, Spain, and the Denver Public Health Department (Denver, Colo.) were analyzed with the two probes. Both probes identified the same isolates in the same clusters when the fingerprints had six or more copies of IS6110. Analysis of isolates with unique IS6110 fingerprints demonstrated that they were unique with pTBN12. The pTBN12 probe had greater discriminating power in isolates having five or fewer copies of IS6110. Forty-seven isolates from Denver having fewer than five copies of IS6110 which were grouped in 11 clusters with identical fingerprint patterns were subdivided into 35 different patterns by pTBN12. Isolates with IS6110 fingerprints with more than six copies of IS6110 that differed from one another by only one or two hybridizing bands were analyzed with pTBN12. Most of these sets of isolates demonstrated identical patterns with pTBN12. However, some exceptions were observed, suggesting that those having nearly identical IS6110 patterns should not necessarily be included in the same cluster. Since IS6110 provides more polymorphism in the fingerprint, it is most useful in identifying isolates with unique fingerprint patterns and those in clusters in which the isolates contain six or more copies of the insertion. However, it is necessary to employ a secondary probe, such as pTBN12, to discriminate isolates with five or fewer copies of IS6110 and those with similar but not identical IS6110 patterns. PMID- 8727888 TI - Most Corynebacterium xerosis strains identified in the routine clinical laboratory correspond to Corynebacterium amycolatum. AB - A comprehensive study was performed on 25 bacterial clinical isolates originally identified as Corynebacterium xerosis. Three reference strains of C. xerosis were also included in the study. On the basis of a variety of phenotypic characteristics tested, all strains could be divided into two separate clusters: reference strains ATCC 373 (the type strain of C. xerosis) and ATCC 7711 showed yellow-pigmented, dry, rough colonies, fermented 5-keto-gluconate, exhibited strong leucine arylamidase and alpha-glucosidase activities, produced lactate as the major end product of glucose metabolism, were susceptible to most of the 19 antimicrobial agents tested, and showed an inhibition zone around disks containing the vibriocidal compound O/129. In contrast, the remaining 26 strains including reference strain NCTC 7243 as well as all clinical isolates formed white-grayish, dry, slightly rough colonies, did not ferment 5-keto-gluconate, exhibited only weak leucine arylamidase and no alpha-glucosidase activity, produced large amounts of propionic acid as the end product of glucose metabolism, and were resistant to most antimicrobial agents tested, including O/129. Chemotaxonomic (cellular fatty acids, mycolic acids, and G+C content) and molecular genetic (16S rRNA gene sequence) investigations revealed that the strains of the second cluster unambiguously belonged to the species C. amycolatum. Our data suggest that most strains reported in the literature as C. xerosis are probably misidentified and correspond to C. amycolatum. PMID- 8727889 TI - Random amplified polymorphic DNA typing of Pseudomonas aeruginosa isolates recovered from patients with cystic fibrosis. AB - Pseudomonas aeruginosa isolates recovered from chronically colonized patients with cystic fibrosis (CF) are phenotypically different from those collected from other patients or from the environment. To assess whether alterations in motility, mucoidy, and serum susceptibility represented an adaptation to chronic infection or replacement by a new strain, sequential P. aeruginosa isolates of known phenotype collected from 20 CF patients were typed by random amplified polymorphic DNA (RAPD) analysis. A total of 35 RAPD strain types were found among 385 isolates from 20 patients, and only two patients had P. aeruginosa strains of the same RAPD fingerprint. Eight strain pairs representative of the first eight RAPD types were also analyzed by SpeI macrorestriction followed by pulsed-field gel electrophoresis (PFGE); the strain types found by both fingerprinting techniques correlated exactly. In 11 of 20 patients, the RAPD types of serial P. aeruginosa isolates remained stable despite alterations in isolate motility, colonial morphology, and lipopolysaccharide phenotype. However, in isolates collected from one CF patient, a single band change in RAPD fingerprint and CeuI PFGE profile correlated with the appearance of an RpoN mutant phenotype, suggesting that the altered phenotype may have been due to a stable genomic rearrangement. Secretion of mucoid exopolysaccharide, loss of expression of RpoN dependent surface factors, and acquisition of a serum-susceptible phenotype in P. aeruginosa appear to evolve during chronic colonization in CF patients from specific adaptation to infection rather than from acquisition of new bacterial strains. PMID- 8727890 TI - Genomic analysis of Mycobacterium bovis and other members of the Mycobacterium tuberculosis complex by isoenzyme analysis and pulsed-field gel electrophoresis. AB - Initially, multilocus enzyme electrophoresis was used to examine genetic relationships among 63 isolates of Mycobacterium bovis and 13 other members of the M. tuberculosis complex. The isolates were divided into five electrophoretic types, with a mean genetic diversity of 0.1. The strains were genetically homogenous, indicating that members of the complex were closely related. This supported the suggestion that they should be considered as subspecies of a single species. Pulsed-field gel electrophoresis (PFGE) was then used to differentiate these isolates, as well as 59 additional isolates of M. bovis from different parts of the world. PFGE differentiated these strains into 63 patterns (53 patterns for M. bovis). Isolates of M. bovis from Western Australia (n = 46) were more homogenous than isolates from other regions. Eight strains were identified in that state, and one predominantly bovine strain was isolated from two human beings and a feral pig. Although M. bovis isolates from different parts of the world had distinct DNA patterns, some were very similar. PFGE is a highly discriminatory technique for epidemiological studies of bovine tuberculosis. For example, it allowed differentiation between isolates of M. bovis cultured from animals in separate outbreaks of tuberculosis, it suggested the transmission of infection between certain properties, and it demonstrated the existence of multiple infections with different strains at certain farms. PMID- 8727891 TI - Preservation of Neisseria gonorrhoeae at -20 degrees C. AB - To explore the feasibility of preserving Neisseria gonorrhoeae at -20 degrees C, we studied its viability quantitatively and qualitatively for 12 and 18 months, respectively, in the following media: a gelatin-based medium used mainly to prepare dried gelatin discs (S. Yamai, Y. Obara, T. Nikkawa, Y Shimoda, and Y. Miyamoto, Br. J. Vener. Dis. 55:90-93, 1979), a simplified version (LSPQ preservation medium), and Trypticase soy broth with 10% (vol/vol) glycerol, a medium commonly used for preservation at -70 degrees C. The latter was studied for 4 months only. Four reference strains and two clinical isolates of N. gonorrhoeae were used. The storage temperature was rigorously preadjusted and monitored at -20 +/- 1 degree C during the entire project. After 12 months of storage, all strains remained viable in both gelatin-based media, whereas a significant loss of viability was observed in Trypticase soy broth-10% glycerol after only 4 months. After 18 months, five strains were still viable in both gelatin-based media and no significant difference was observed between antimicrobial susceptibility results and those of the original strains preserved at -70 degrees C. On the basis of these results, we believe that LSPQ preservation medium represents a good alternative for the storage of N. gonorrhoeae at -20 degrees C for at least a year. Furthermore, it is easy to prepare and use and can by stored at 4 to 8 degrees C for a year prior to use. PMID- 8727892 TI - Evaluation of three commercial enzyme immunoassays compared with the 13C urea breath test for detection of Helicobacter pylori infection. AB - The diagnostic significance of the serological detection of antibodies to Helicobacter pylori has been established by numerous investigators. Reports of the clinical reliabilities of commercial enzyme immunoassay (EIA) kits available for this purpose vary as a result of the different H. pylori antigen sources and reference methods used. The 13C urea breath test (UBT) has been shown to be an extremely accurate and reliable method of detecting H. pylori infection. We used the 13C urea breath test as the confirmatory method for H. pylori status to evaluate three commercially available EIA kits designed to detect immunoglobulin G antibodies to H. pylori. These kits were the HM-CAP EIA kit (Enteric Products, Inc.), the PYLORI STAT EIA kit (BioWhittaker, Inc.), and the G.A.P. kit (Bio-Rad Laboratories/Biomerica, Inc.). The evaluations were performed in a double-blind manner with samples from 473 clinically characterized patients. This group included patients with symptomatic gastrointestinal disorders as well as nonsymptomatic volunteers. The sensitivities of the kits were as follows: HM-CAP, 98.4%; PYLORI STAT, 99.2%; and G.A.P., 100%. The specificities were as follows: HM-CAP, 96.4%; PYLORI STAT, 90.1%; and G.A.P., 26.0%. Although the HM-CAP and PYLORI STAT kits performed comparably, the G.A.P. test yielded significantly more false-positive results and an unacceptably high number of indeterminate results. PMID- 8727893 TI - Comparison of PCR-based approaches to molecular epidemiologic analysis of Clostridium difficile. AB - Representative isolates of the 10 serogroups of Clostridium difficile and 39 clinical isolates (30 toxigenic and 9 nontoxigenic), including 5 isolates from a confirmed nosocomial outbreak, were analyzed by using two previously described arbitrary-primer PCR (AP-PCR) molecular typing methodologies (AP-PG05 and AP ARB11) and PCR ribotyping. The two AP-PCR methods investigated gave comparable results; AP-PG05 and AP-ARB11 identified 8 and 7 groups among the serogroup isolates and classified the clinical isolates into 21 and 20 distinct groups, respectively. PCR ribotyping also identified 8 unique groups among the serogroup isolates but classified the clinical isolates into 23 groups. In addition, when results obtained by the PCR methods were compared with typing data generated by pulsed-field gel electrophoresis (PFGE), PCR ribotyping and PFGE were found to be in agreement for 83% (29 of 35) of isolates typeable by both techniques while AP PG05 was in agreement with PFGE for 60% (20 of 33) and AP-ARB11 was in agreement with PFGE for only 44% (17 of 36). These results indicate that PCR ribotyping is a more discriminatory approach than AP-PCR for typing C. difficile and, furthermore, that this technique generates results that are in higher concordance with those obtained by using an established method for differentiating isolates of this organism on a molecular level than are results generated by using AP-PCR. PMID- 8727894 TI - Bartonella (Rochalimaea) quintana infection in a seronegative hemodialyzed patient. AB - Bartonella quintana is a reemerging pathogen responsible for trench fever, endocarditis, bacteremia, and bacillary angiomatosis. We previously reported the first case of a patient with B. quintana-induced chronic adenomegaly, and here we present a report on a second patient. A hemodialyzed patient with Sjogren's syndrome presented with mediastinal adenomegalies and secondary pancytopenia. All diagnostic investigations remained negative, except that a Bartonella-like microorganism was isolated from a bone marrow biopsy. The isolate was identified as B. quintana by a specific mouse polyclonal antibody and by determination of a partial gltA (citrate synthase-encoding) gene and 16S rRNA gene sequences. DNA of the pathogen was also detected in the adenomegaly and in the serum of the patient by PCR amplification of the gltA gene. Anti-B. quintana antibodies were never detected in the patient's serum throughout the 12-month follow-up but were detected in the serum of the patient's cat. The patient's outcome was favorable after treatment with gentamicin. Chronic adenomegaly in seronegative patients is a new clinical entity due to B. quintana. PMID- 8727895 TI - Role of stool screening tests in diagnosis of inflammatory bacterial enteritis and in selection of specimens likely to yield invasive enteric pathogens. AB - The Leuko-Test yielded a negative predictive value of 98.4% when it was used to screen 325 patients for inflammatory bacterial enteritis and a negative predictive value of 99.4% when it was used to screen 416 stool specimens for those from which enteric pathogens would likely be recovered when cultured. Neither microscopy for fecal leukocytes nor an assay for fecal occult blood, alone or in combination, allowed for the reliable detection of invasive bacterial enteritis or the reliable selection of specimens for culture. When positive in the Leuko-Test, specimens collected from patients after the third day of hospitalization did not yield enteric pathogens when the specimens were cultured, and specimens collected from inpatients within the first 3 days of hospitalization or from outpatients did not contain Clostridium difficile toxin A. As a screening test, the Leuko-Test has the ability to generate rapidly a result which can support the presumptive diagnosis of inflammatory bacterial enteritis or which can be used to determine the suitability of stool specimens for bacteriologic culture. PMID- 8727896 TI - A three-center European external quality control study of PCR for detection of cytomegalovirus DNA in blood. AB - The presence of cytomegalovirus (CMV) in the blood has important consequences for patient management, and an external quality control study of its detection by the PCR was conducted by the Infectious Disease Working Party of the European Group for Blood and Marrow Transplantation. Forty-eight coded peripheral blood samples from bone marrow transplant recipients were processed in parallel in three European centers by using the routine in-house PCR assay. Protocols varied in choice of primers, specificity and amplificability controls, and sample processing. Results for 38 of 47 samples agreed, 35 being negative and 3 positive. Of the 12 samples reported as positive by a least one center, only 3 were found to be positive by all three centers, 1 was found to be positive by two centers, and the remaining 8 were found to be positive by one center only. The nine discrepant samples appeared to contain around 1,000-fold less viral DNA than the three concordant positive samples. CMV detection was affected both by the number of leukocytes from which DNA was extracted and by the number of cell equivalents added per PCR. External quality control schemes for CMV PCR are clearly necessary in order to compare data from different centers, and recommendation for standardizing the PCR detection of CMV in blood leukocytes are made. PMID- 8727897 TI - Comparison of PCR and microscopic methods for detecting Trypanosoma cruzi. AB - The diagnosis of acute infection with Trypanosoma cruzi, the protozoan parasite that causes Chagas' disease, is generally made by detecting parasites by microscopic examination of fresh blood. Although highly specific, this approach often lacks sensitivity. Several years ago, PCR assays for the detection of T. cruzi were described, but the sensitivities and specificities of these tests have not yet been defined precisely. In the present study, we first compared the sensitivities of PCR methods that differ in sample processing as well as in the target sequences that are amplified. Then, we challenged eight mice with T. cruzi, and on 31 days over a 380-day period, we compared the ability of the PCR method with the highest sensitivity to detect parasites in blood with that of microscopic examination. During the acute phase of the infections, parasites were detected on average 3.9 days earlier by the PCR method than by microscopy. Furthermore, the infected mice were consistently positive by the PCR method during the chronic phase, while parasites were intermittently detected by microscopic examination during that period. Overall, among the 248 comparisons, in 84 the PCR method was positive and no parasites were seen by microscopic examination, whereas the reverse was true in only 1 case, a difference that is highly significant. These findings suggest that this approach should be in patients suspected of having acute Chagas' disease. Moreover, the higher sensitivity of the PCR method observed in both the acute and chronic phases of the T. cruzi infections in the mice that we studied indicates that this approach should be useful in evaluating experimental drugs in T. cruzi-infected laboratory animals. PMID- 8727898 TI - Novel BOX repeat PCR assay for high-resolution typing of Streptococcus pneumoniae strains. AB - Typing data obtained by specifically targeting a single, high-stringency PCR at the pneumococcal BOX repeat element for 28 strains of Streptococcus pneumoniae completely corroborated the resolutions attained by five genotypic procedures as described by Hermans et al. (P.W.M. Hermans, M. Sluijter, T. Hoogenboezem, H. Heersma, A. van Belkum, and R. de Groot, J. Clin. Microbiol. 33:1606-1612, 1995). All pairs of strains, except one, derived from both the cerebrospinal fluid and blood of the same individual were shown to be identical. Moreover, other, epidemiologically unrelated isolates were demonstrated to be unique. Considering the combined data from the five typing techniques applied previously as the "gold standard," the single BOX PCR test demonstrated excellent resolving powers while maintaining epidemiological linkage. PMID- 8727899 TI - Performance of Meridian ImmunoCard Mycoplasma test in a multicenter clinical trial. AB - Serology is the principal laboratory method used to diagnose Mycoplasma pneumoniae infection. Meridian Diagnostics has developed the ImmunoCard Mycoplasma kit, a 10-min card-based enzyme-linked immunosorbent assay (ELISA) designed to detect immunoglobulin M (IgM) antibodies to M. pneumoniae. We compared the ImmunoCard with two M. pneumoniae IgM-specific assays (immunofluorescence assay [IFA] and ELISA) and a standard complement fixation (CF) procedure using 896 specimens submitted to clinical laboratories for M. pneumoniae serology. Equivocal results obtained by CF, IFA, or ELISA were resolved by testing with an additional method or by reviewing patient chart information. The ImmunoCard had sensitivities ranging from 74% compared with the ELISA to 96% compared with CF results with IFA. ImmunoCard specificities ranged from 85% compared with the IgM-specific ELISA to 98% compared with IgM-specific IFA results resolved with clinical chart review. We also compared the ImmunoCard results with consensus results of 694 specimens tested on at least two non ImmunoCard methods because of the lack of a "gold standard" for M. pneumoniae serology. Overall, the ImmunoCard Mycoplasma IgM assay had 90% sensitivity, 93% specificity, and 92% agreement with the consensus results. The ImmunoCard is technically less complex and requires less equipment that the three other assays. Our results indicate that the ImmunoCard Mycoplasma IgM assay is a valid and simple procedure which can reduce technologist time (and, thus, labor cost) and turnaround time for laboratories analyzing small numbers of specimens (< 10 per batch) submitted for IgM anti-M. pneumoniae testing. PMID- 8727900 TI - Detecting bunyaviruses of the Bunyamwera and California serogroups by a PCR technique. AB - Many bunyaviruses of the Bunyamwera and California serogroups are medically important human pathogens. The development of an effective technique to detect the viruses by using molecular biologic tools, such as PCR, improves not only clinical diagnosis but also virologic surveillance of mosquito vectors in the field. In this study, we evaluated eight pairs of primers for reactivity with 44 viruses of the genus Bunyavirus, using a reverse transcriptase PCR technique. With a pair of serogroup-specific primers we designed, all viruses of the serogroups tested could be detected. Further, virus-specific primer pairs were identified for California encephalitis virus, Jamestown Canyon virus, La Crosse virus, and snowshoe hare virus for use in North America. Using this technique, we could detect one La Crosse virus-infected mosquito in a pool of 100 mosquitoes with undetectable plaque titers. PMID- 8727901 TI - Molecular characterization of Vibrio cholerae O1 strains isolated during cholera outbreaks in Guinea-Bissau. AB - In the present study, 19 strains of Vibrio cholerae O1 biotype El Tor isolated during outbreaks of cholera in Guinea-Bissau in 1987, 1994, and 1995 were characterized to investigate a possible epidemiological relationship among the isolates. On the basis of ribotyping with the restriction enzyme BglI, 5 strains isolated in 1987 showed two closely related ribotypes, while 14 strains isolated in 1994 and 1995 showed the same ribotype that was distinct from the ribotypes of strains isolated in 1987. Southern blot hybridization of BglI-digested genomic DNA with a cholera toxin probe demonstrated that the strains isolated in 1987 showed an identical cholera toxin genotype, whereas O1 strains isolated in 1994 and 1995 showed the same genotype that was distinct from the genotype of strains isolated in 1987. These results were supported by the results of antibiotic susceptibility testing, in which strains isolated in 1987 showed resistance to polymyxin B only, while each of the strains from 1994 and 1995 showed resistance to polymyxin B, trimethoprim-sulfamethoxazole, and the vibriostatic agent O/129. Although our results are based on a limited number of V. cholerae O1 strains, they suggest that the epidemic in Guinea-Bissau in 1994 and 1995 was due to the introduction of a new strain to the country. PMID- 8727902 TI - Validation of use of whole-cell repetitive extragenic palindromic sequence-based PCR (REP-PCR) for typing strains belonging to the Acinetobacter calcoaceticus Acinetobacter baumannii complex and application of the method to the investigation of a hospital outbreak. AB - Acinetobacter spp. are being reported with increasing frequency as causes of nosocomial infection. In order to identify reservoirs of infection as quickly as possible, a rapid typing method that can differentiate epidemic strains from environmental and nonepidemic strains is needed. In 1993, a cluster of Acinetobacter baumannii isolates from five patients in the adult intensive therapy unit of our tertiary-care teaching hospital led us to develop and optimize a rapid repetitive extragenic palindromic sequence-based PCR (REP-PCR) typing protocol for members of the Acinetobacter calcoaceticus-A. baumannii complex that uses boiled colonies and consensus primers aimed at repetitive extragenic palindromic sequences. Four of the five patient isolates gave the same REP-PCR typing pattern as isolates of A. baumannii obtained from the temperature probe of a Bennett humidifier; the fifth isolate had a unique profile. Disinfection of the probe with 70% ethanol, as recommended by the manufacturer, proved ineffective, as A. baumannii with the same REP-PCR pattern was isolated from it 10 days after cleaning, necessitating a change in our decontamination procedure. Results obtained with REP-PCR were subsequently confirmed by ribotyping. To evaluate the discriminatory power (D) of REP-PCR for typing members of the A. calcoaceticus-A. baumannii complex, compared with that of ribotyping, we have applied both methods to a collection of 85 strains that included representatives of six DNA groups within the complex. Ribotyping using EcoRI digests yielded 53 patterns (D = 0.98), whereas 68 different REP-PCR patterns were observed (D = 0.99). By computer-assisted analysis of gel images, 74 patterns were observed with REP-PCR (D = 1.0). Overall, REP-PCR typing proved to be slightly more discriminatory than ribotyping. Our results indicate that REP PCR typing used boiled colonies is a simple, rapid, and effective means of typing members of the A. calcoaceticus-A. baumannii complex. PMID- 8727903 TI - Identification of bovine Neospora parasites by PCR amplification and specific small-subunit rRNA sequence probe hybridization. AB - Neospora is a newly recognized genus of pathogenic coccidia, closely related to Toxoplasma gondii, that can cause abortion or congenital disease in a variety of domestic animal hosts. On the basis of the small-subunit rRNA gene sequences of Neospora spp. and other apicomplexa coccidia, oligonucleotide primers COC-1 and COC-2 were used for PCR amplification of conserved sequences of approximately 300 bp in size. A Neospora-specific chemiluminescent probe hybridized to Southern blots of amplification products from Neospora DNA but not to Southern blots with amplified DNA from the other coccidian parasites tested. A Toxoplasma-specific probe whose sequence differed from that of the probe for Neospora spp. by a single base pair was used to distinguish these parasites by specific Southern blot hybridization. The PCR system detected as few as one Neospora tachyzoite in the culture medium or five tachyzoites in samples of whole blood or amniotic fluid spiked with Neospora parasites. In addition, Neospora PCR products were successfully amplified from whole blood and amniotic fluid samples of experimentally infected bovine and rhesus macaque fetuses. These results indicate that this PCR and probe hybridization system could be a valuable adjunct to serology and immunohistochemistry for the diagnosis of Neospora infections in bovine or primate fetuses. PMID- 8727904 TI - Sequence capture-PCR improves detection of mycobacterial DNA in clinical specimens. AB - The rapid identification of mycobacterial DNA in clinical samples by PCR can be useful in the diagnosis of tuberculous infections, but several large studies have found that the sensitivity of this approach is not better than that of culture. In order to improve the sensitivity of detection of mycobacterial DNA in clinical specimens from patients with paucibacillary forms of tuberculosis, we have developed a procedure permitting the specific capture of mycobacterial DNA in crude samples prior to amplification, thereby concentrating the target sequences and removing irrelevant DNA and other potential inhibitors of the amplification reaction (sequence capture-PCR). By using this approach to capture and amplify two different sequences specific for organisms of the Mycobacterium tuberculosis complex (IS6110 and the direct repeat region), it was possible to detect as little as one genome of mycobacterial DNA in samples containing up to 750 micrograms of total DNA, representing a 10- to 100-fold increase in sensitivity compared with that obtained by purifying total DNA prior to amplification. Detection of the IS6110 sequence in pleural fluid samples from patients with tuberculous pleurisy by sequence capture-PCR gave positive results in 13 of 17 cases, including 3 of 3 culture-positive samples and 10 of 14 culture-negative samples. In contrast, when total DNA was purified from these samples by adsorption to a silica matrix prior to amplification, only the three culture positive samples were positive by PCR. The sensitivity of detection of the direct repeat sequence in these samples by sequence capture-PCR was similar to that of IS6110 and, in addition, permitted immediate typing of the strains from some patients. We conclude that sequence capture-PCR improves the sensitivity of detection of mycobacterial DNA in paucibacillary samples. This approach should be useful in detecting rare target sequences from organisms implicated in other pathologic processes. PMID- 8727905 TI - Restriction fragment length polymorphism of flagellin genes of Campylobacter jejuni and/or C. coli isolates from Egypt. AB - The conservation of flagellin genes from thermophilic Campylobacter spp. strains isolated in Egypt was evaluated by a restriction fragment length polymorphism (RFLP) assay. The flaA and flaB genes were amplified from 59 independent clinical isolates and digested with EcoRI and PstI, and the resulting patterns were compared with each other and with previously described RFLP groups. The results indicate that the isolates fell into 14 groups for flaA and 11 groups for flaB, 9 of which have been described, and that considerable genetic variability exists among isolates belonging to the same LIO serogroup. In most cases, the flaB gene displayed the same RFLP pattern as that of the flaA gene of the same strain, although some variability was observed. The data suggest that more variability of flagellin genes exists within the LIO serogroups common to Campylobacter field isolates from Egypt than has previously been reported for North American isolates. PMID- 8727906 TI - Outbreak of histoplasmosis among employees in a paper factory--Michigan, 1993. AB - In December 1993, four reported cases of histoplasmosis among employees in a Michigan pulp paper factory prompted an investigation. A cohort of employees was surveyed to identify additional cases. A case of acute histoplasmosis was defined as an influenza-like illness in a plant employee with the onset of illness during October or November 1993 and laboratory evidence of recent infection with Histoplasma capsulatum. Among the 96 employees surveyed, 18 persons met the case definition; all of these had illness onset during the last week of October in 1993. The attack rate among maintenance employees (16 of 53 [30%]) was much greater than that among nonmaintenance employees (2 of 43 [5%]) (relative risk = 6.5; 95% confidence interval = 1.6 to 26.7; P = 0.003). On October 22, a dry, windy day, one maintenance worker swept bird guano, approximately 10 cm deep, from an adjacent roof < 20 m from the maintenance building. The disturbance of the bird guano was the likely event which caused this outbreak. H. capsulatum remains an important pathogen among immunocompetent hosts. We recommended that the plant authorities (i) discourage birds from roosting at the facility and (ii) use safe procedures for the cleanup and disposal of soil contaminated with bird droppings. PMID- 8727907 TI - Detection of Yersinia enterocolitica serogroup O:3 by a PCR method. AB - Yersinia enterocolitica is the etiologic agent of a range of clinical situations in humans, but only a small number of serotypes are involved. Among these, Y. enterocolitica O:3 is the most frequently implicated. A PCR method was developed to detect Y. enterocolitica O:3. For this purpose, two pairs of primers were designed to amplify two fragments of the rfb cluster of Y. enterocolitica O:3: a 253-bp fragment of the rfbB gene and a 405-bp fragment of the rfbC gene. A specific detection was obtained only with rfbC primers, which yielded a PCR product of the expected size exclusively with pathogenic Y. enterocolitica of serotype O:3. This pair of primers was combined with the ail, inv, and virF primers previously described (H. Nakajima, M. Inoue, T. Mori, K.-I. Itoh, E. Arakawa, and H. Watanabe, J. Clin. Microbiol. 30:2484-2486, 1992) to allow both the detection and the differentiation between Y. pseudotuberculosis, pathogenic Y. enterocolitica of serotype O:3 and other pathogenic Y. enterocolitica. PMID- 8727908 TI - Immunohistochemical detection of prion protein in lymphoid tissues of sheep with natural scrapie. AB - The scrapie-associated form of the prion protein (PrPSc) accumulates in the brain and lymphoid tissues of sheep with scrapie. In order to assess whether detecting PrPSc in lymphoid tissue could be used as a diagnostic test for scrapie, we studied the localization and distribution of PrPSc in various lymphoid tissues collected at necropsy from 55 sheep with clinical scrapie. Samples collected from the spleen, palatine tonsil, ileum, and five different lymph nodes were immunohistochemically stained for PrPSc. PrPSc was found to be deposited in a reticular pattern in the center of both primary and secondary lymphoid follicles. In addition, granules of PrPSc were seen in the cytoplasm in macrophages associated with the lymphoid follicles. In 54 (98%) of the 55 scrapie-affected sheep, PrPSc was detected in the spleen, retropharyngeal lymph node, mesenteric lymph node, and the palatine tonsil. However, only in the palatine tonsils was PrPSc present in a consistently high percentage of the lymphoid follicles. PrP was not detected in any of the lymphoid tissues of 12 sheep that had no neurohistopathological signs of a scrapie infection. We conclude that the tonsils are the best-suited lymphoid tissue to be biopsied for the detection of PrPSc in the diagnosis of clinical scrapie in living sheep. PMID- 8727909 TI - Metronidazole and clarithromycin resistance in Helicobacter pylori determined by measuring MICs of antimicrobial agents in color indicator egg yolk agar in a miniwell format. The Gastrointestinal Physiology Working Group of Universidad Peruana Cayetano Heredia and the Johns Hopkins University. AB - Resistance of Helicobacter pylori to metronidazole often causes failure of commonly used combination drug treatment regimens. We determined the MICs of metronidazole and clarithromycin against 18 H. pylori strains from Peru using tetrazolium egg yolk (TEY) agar. The MIC results obtained by agar dilution with petri dishes were compared with the results found through a miniwell format. The results of the two protocols for measuring drug susceptibility differed by no more than 1 dilution in all cases. On TEY agar, bright-red H. pylori colonies were easy to identify against a yellow background. Sixty-one percent (11 of 18) of the strains were resistant to metronidazole (MIC, > or = 4 micrograms/ml) and 50% (9 of 18) were resistant to clarithromycin (MIC, > or = 0.125 micrograms/ml), whereas none (0 of 5) of the strains tested were resistant to tetracycline (MIC, > or = 1 micrograms/ml). Thus, the prevalence of metronidazole and clarithromycin resistance in Peru is higher than that in developed regions of the world. The miniwell plate with TEY agar allows easy H. pylori colony identification, requires about one-third less of the costly medium necessary for petri dish assaying, conserves space, and yields MICs equivalent to those with agar dilution in petri dishes. PMID- 8727910 TI - Typing Candida albicans oral isolates from human immunodeficiency virus-infected patients by multilocus enzyme electrophoresis and DNA fingerprinting. AB - A total of 189 Candida albicans isolates have been typed by multilocus enzyme electrophoresis. The results obtained confirm the clonal mode of reproduction of C. albicans. The C. albicans populations found in the oropharynx of human immunodeficiency virus (HIV)-infected patients, in the oropharynx of healthy carriers, or in association with invasive candidiasis could not be distinguished. No clone or group of clones could be associated with the appearance of clinical disorders or with a reduced in vitro susceptibility to the antifungal agent fluconazole. Multiple and sequential oral isolates from 24 HIV-infected patients were also typed by restriction enzyme analysis with the enzymes EcoRI and HinfI and by use of the Ca3 repetitive probe. The results obtained by the combination of all three typing methods show that all but one patient each carried a unique major C. albicans clone in their oropharynx. The 21 patients with sequential isolates had the same C. albicans clones in their throats during recurrent oropharyngeal candidiasis episodes, independently of clinical status or of changes of in vitro susceptibility to fluconazole. Finally, several isolates of the same C. albicans clone found simultaneously in the oropharynx of a patient may present different levels of susceptibility to fluconazole. PMID- 8727911 TI - Antagonistic effect of oral bacteria towards Treponema denticola. AB - This study was designed to isolate oral bacteria exhibiting antagonism towards Treponema denticola and to characterize the inhibitory activity. Eleven bacterial isolates obtained from subgingival sites and identified as either Staphylococcus aureus or Streptococcus mutans were found to inhibit the growth of T. denticola. When the activity spectra of these isolates were analyzed, two additional periodontopathogens (Porphyromonas gingivalis and Prevotella intermedia) were found to be affected, whereas most gram-positive bacteria were not. Strains of S. aureus produce a bacteriocin-like inhibitory substance (heat stable and protease sensitive), whereas the inhibitory effect of S. mutans appears to be related to the production of lactic acid. The negative interactions reported in this paper may govern population shifts observed in subgingival sites. PMID- 8727912 TI - Concordance of clinical and environmental isolates of Cryptococcus neoformans var. gattii by random amplification of polymorphic DNA analysis and PCR fingerprinting. AB - Sixty-one clinical and forty-nine environmental isolates of Cryptococcus neoformans var. gattii from Australia and the United States were analyzed by random amplification of polymorphic DNA (RAPD), using 12- to 22-mer primers in pairs, and/or PCR fingerprinting with a single primer derived from the microsatellite core sequence of the wild-type phage M13 (5' GAGGGTGGCGGTTCT 3'). Three major genetic profiles were identified by both typing techniques. A single RAPD profile (VGI) predominated among clinical isolates (44 of 48, 92%) and isolates from host eucalypts (45 of 45, 100%) from Australia. Of the 94 Australian isolates, 4 (3 clinical and 1 environmental) were assigned to profile VGII; 2 of these were recovered from patients and one was recovered from plant debris from Western Australia. Only one Australian clinical isolate was assigned to profile VGIII. A different distribution of RAPD profiles (four VGIII, two VGII, and one VGI) was found among four clinical and three environmental isolates from the United States. RAPD profiles of 8 of the 101 isolates studied revealed minor genetic variants, 4 of profile VGI and 4 of profile VGII. Genetic concordance between the majority of clinical and environmental isolates in Australia is consistent with the hypothesis that human disease is acquired from exposure to host eucalypts. Profiles of clinical isolates were independent of body site of infection, and profiles of all isolates were stable over time. Analysis by PCR fingerprinting confirmed the RAPD results. A second RAPD profile (VGII) was associated with infection in southwest Western Australia, where the two host eucalypts do not occur naturally. This raises the possibility of an alternative and as yet unidentified natural habitat of C. neoformans var. gattii. Our results indicate that RAPD analysis is a sensitive and useful method for investigating environmental sources of human infection with this biotype. PMID- 8727913 TI - Natural environmental sources of Cryptococcus neoformans var. gattii. AB - We sought evidence for new environmental sources of Cryptococcus neoformans var. gattii by random amplification of polymorphic DNA (RAPD) analysis of isolates from 29 animals with a restricted territorial range in five Australian states. Twenty-three of the 29 isolates and 45 of 45 eucalypt isolates tested previously exhibited one RAPD profile, VGI. RAPD profile VGII was identified in 6 of 17 isolates from domesticated species but in none of 12 native species. Four VGII isolates originated from an area of Western Australia with no natural stands of known eucalypt host, indicating the existence of at least one unrecognized natural source of C. neoformans var. gattii. PMID- 8727914 TI - Nosocomial CDC group IV c-2 bacteremia: epidemiological investigation by randomly amplified polymorphic DNA analysis. AB - The CDC group IV c-2 bacterium is a gram-negative bacillus rarely isolated from clinical specimens. This organism caused catheter-related bacteremia in five immunocompromised children hospitalized in two distinct wards of our institution between November 1993 and October 1994. Three patients recovered on empiric antibacterial chemotherapy combining ceftazidime and amikacin, and a fourth patient required imipenem instead of ceftazidime. The fifth patient recovered without treatment. Catheter removal was never necessary. The randomly amplified polymorphic DNA technique with three different primers was applied to nine isolates recovered by culturing blood from the five children and showed that all of the patients harbored isolates of the same genotype. The source of the outbreak could not be determined. PMID- 8727915 TI - Identification and differentiation of Mycobacterium avium and M. intracellulare by PCR. AB - Known DNA sequences coding for the 16S rRNAs of 14 slowly growing Mycobacterium species were analyzed. Three sets of primers were synthesized: MAV and MIN, for M. avium and M. intracellulare, respectively, and MYCOB, for the slowly growing mycobacteria. Whole-cell DNAs of 14 reference species were extracted and amplified by PCR with the MYCOB, MAV, and MIN primers. The MYCOB primer amplified a 0.9-kb segment from the DNAs of all 14 species. The MAV and MIN primers each amplified one highly specific 1.3-kb segment from the homologous DNA, respectively. DNAs from each of 10 clinical isolates of M. avium and M. intracellulare identified by conventional methods were amplified with the MYCOB as well as the MAV and MIN primers; 9 of 10 isolates of each species were identified with their respective primers. One isolate of M. intracellulare was subsequently found to have been mislabeled. One isolate designated M. avium reacted only with the MYCOB primer. The hypervariable region of this strain was shown by DNA sequence analysis to be distinct from all known 16S rRNA sequences of Mycobacterium spp. Our data indicate that the currently identified M. avium-M. intracellulare complex includes strains genetically diverse from M. avium and M. intracellulare. PMID- 8727916 TI - Bovine adenovirus type 10 identified in fatal cases of adenovirus-associated enteric disease in cattle by in situ hybridization. AB - A severe, naturally occurring enteric disease of cattle in which adenovirus inclusions are present in the intestinal vascular endothelium has been recognized in several countries; three different adenovirus serotypes have been isolated from affected animals. An in situ hybridization technique for the detection of bovine adenoviral DNA was developed and applied to tissue from 13 cattle in Northern Ireland diagnosed to have the disease. Bovine adenovirus serotype 10 (BAV-10) was identified in the vascular inclusions of all cattle, providing strong evidence that adenoviral enteric vascular disease in cattle is associated with this serotype. The existence of BAV-10 has only recently been recognized. The first molecular biology-based technique for the diagnosis of BAV-10 infection is described. The animals in the present study are the first in which BAV-10 has had a confirmed role in a pathologic process. PMID- 8727917 TI - Misidentification of toxigenic Corynebacterium diphtheriae as a Corynebacterium species with low virulence in a child with endocarditis. AB - A 6-year-old boy presented to a university hospital in Malaysia with infective endocarditis complicating cyanotic congenital heart disease. Blood cultures showed a gram-positive, aerobic, coryneform-like bacillus identified by the hospital laboratory as Corynebacterium xerosis, but a reference laboratory identified the organism as a toxigenic strain of Corynebacterium diphtheriae. The two laboratories concurred on all biochemical test results except for sucrose fermentation. PMID- 8727918 TI - Comparative susceptibilities of human embryonic fibroblasts and HeLa cells for isolation of human rhinoviruses. AB - The recovery of human rhinovirus (HRV) from nasal washings and nasal and pharyngeal swabs from volunteers with naturally acquired colds was compared in different cell types. Human embryonic lung fibroblast (HELF) strain WI-38 (sensitivity, 61 to 84%) and HeLa-I, an HRV-susceptible HeLa clone (sensitivity, 86 to 94%), were the most sensitive cell types used. HELF-WI-38 cells showed a cytopathic effect earlier than the other cells used, and the different strains of HRV-susceptible HeLa cells varied in their sensitivities for HRV isolation. HRV was detected in a single cell type in 20 to 35% of the positive samples, suggesting that use of a combination of different HRV-susceptible cell lines is the best approach for the recovery of HRV. Although nasal washings tended to yield more HRV isolates than nasal and pharyngeal swabs, the two sampling methods were not found to be significantly different. PMID- 8727919 TI - Multicenter evaluation of broth microdilution method for susceptibility testing of Cryptococcus neoformans against fluconazole. AB - We have developed a microdilution method for measuring the susceptibility of Cryptococcus neoformans to fluconazole. The present study evaluated the interlaboratory agreement of the results for the microdilution method obtained at three different sites and compared this method with the National Committee for Clinical Laboratory Standards M27-P reference method. Excellent interlaboratory agreement among the results obtained at the three sites was achieved with this method (83 and 96% agreement within 1 and 2 log2 dilutions, respectively). An overall agreement of 90% between the microdilution method and the M27-P method was observed, demonstrating the comparability of the two methods. However, there are inherent problems with the M27-P method in relation to measuring C. neoformans susceptibility, including suboptimal growth of the organism in RPMI 1640, a longer incubation period, and a narrow range of MICs. On the basis of these data, the microdilution method tested in this study is recommended for inclusion in the National Committee for Laboratory Standards method for testing the antifungal susceptibility of C. neoformans. PMID- 8727920 TI - Veillonella infections in children. AB - From 1974 to 1994, 2,033 specimens from children were submitted for cultures for anaerobic bacteria. Eighty-three Veillonella spp. were recovered from 83 children (4%). Most Veillonella species were recovered from abscesses, aspiration pneumonias, burns, bites, and sinuses. The infections were polymicrobial in 79 (95%) patients, but in 4 (5%) patients, Veillonella species were recovered in pure culture. The predisposing conditions associated with the recovery of these organisms were previous surgery, malignancy, steroid therapy, foreign body, and immunodeficiency. These data illustrate that Veillonella spp. are found infrequently in children, mostly in association with mixed infections, and are recovered mixed with mouth and bowel flora. PMID- 8727921 TI - Evaluation of commercial latex reagents for identification of O157 and H7 antigens of Escherichia coli. AB - Agglutination reactions obtained with three commercial latex reagents for detecting Escherichia coli O157 antigen (Oxoid Diagnostic Reagents, Hampshire, England; Pro-Lab Inc., Richmond Hill, Ontario, Canada; and Remel Microbiology Products, Lenexa, Kans.) and one for detecting H7 antigen (Remel) were compared with those obtained by standard serologic methods by using the Centers for Disease Control and Prevention (CDC) reference antisera for O157 and H7 antigens. For 159 strains of E. coli and related organisms, the Oxoid, Pro-Lab, and Remel O157 latex reagents each had a sensitivity and specificity of 100% compared with the CDC reference antiserum. For 106 strains of E. coli and related organisms that were not enhanced for motility through semisolid medium, the Remel H7 latex reagent had a sensitivity of 96% and a specificity of 100% compared with the standard tube agglutination method with CDC H7 antiserum. Measures to enhance motility were needed for some strains to detect the H7 antigen. Our findings demonstrate that the commercial latex reagents are good alternatives to standard serologic methods for identifying the O157 and H7 antigens of E. coli. PMID- 8727922 TI - Endocarditis of native aortic and mitral valves due to Corynebacterium accolens: report of a case and application of phenotypic and genotypic techniques for identification. AB - Endocarditis of native aortic and mitral valves due to an organism identified as Corynebacterium accolens developed in a 73-year-old patient without predisposing factors. The organism was identified as C. accolens by biochemical identification, amplified rRNA gene restriction analysis, and DNA-DNA hybridization. This is the first case of C. accolens endocarditis reported, adding to the increasing number of Corynebacterium-related cases of endocarditis. PMID- 8727923 TI - Restriction fragment length polymorphism of the tdh and trh genes in clinical Vibrio parahaemolyticus strains. AB - The restriction fragment length polymorphism of the genes encoding thermostable direct hemolysin (tdh) and thermostable direct hemolysin-related hemolysin (trh) was analyzed for 137 strains of Vibrio parahaemolyticus isolated from specimens from diarrheal patients in Thailand. The HindIII restriction fragment patterns of tdh and trh were grouped into five and four types, respectively. A strong association between the restriction fragment patterns of tdh and trh was observed with V. parahaemolyticus strains. PMID- 8727924 TI - Antimicrobial susceptibilities of Lactococcus lactis and Lactococcus garvieae and a proposed method to discriminate between them. AB - The MICs of antimicrobial agents contained in the SCEPTOR Streptococcus MIC panels (Becton Dickinson Microbiology Systems) were determined for Lactococcus lactis, L. garvieae, and unknown Lactococcus species. Several isolates had reduced susceptibilities to many of the antimicrobial agents contained in the panel. For L. garvieae, the MICs of penicillin and, possibly, cephalothin were higher than for L. lactis, and unlike L. lactis, L. garvieae was resistant to clindamycin, indicating that knowledge of the Lactococcus species causing an infection might influence the choice of antimicrobial therapy. Susceptibility to clindamycin can also be used to differentiate between L. lactis and L. garvieae. PMID- 8727925 TI - Polymorphism in Brucella strains detected by studying distribution of two short repetitive DNA elements. AB - Thirty-four Brucella reference or field strains representing all the species and biovars were studied by repetitive element sequence-based PCR, a PCR using primers complementary to two enterobacterial short repetitive sequences: repetitive extragenic palindromic and enterobacterial repetitive intergenic consensus sequences. All the stains showed a positive amplification, suggesting that the Brucella genome contains such sequences. Repetitive extragenic palindromic PCR was less discriminating than enterobacterial repetitive intergenic consensus PCR in terms of distinguishing strains, but a combination of the two methods was able to distinguish all the isolates, except for some strains belonging to biovars 3 and 9 of Brucella abortus. Repetitive element sequence based PCR appears to be a simple and useful method for the study of brucellosis epidemiology. PMID- 8727926 TI - Identification of group B rotaviruses with short genome electropherotypes from adult cows with diarrhea. AB - Two field strains (BB-RVLV and KD) of group B rotaviruses from adult dairy cows with diarrhea displayed short genome electropherotypes. Gnotobiotic calves inoculated with fecal filtrates of each group B rotavirus developed diarrhea, and only group B rotaviruses or antigens were detected in the feces by immunoelectron microscopy and in intestinal epithelial cells by immunofluorescent staining, respectively. The feces or intestinal contents of the cows and inoculated calves were negative for group A and C rotaviruses by enzyme-linked immunosorbent assay, immunoelectron microscopy, or cell culture immunofluorescence assays. Comparison of the genome electropherotypes of the calf-passaged BB-RVLV and KD strains with the original samples and reference bovine group A, B, and C rotaviruses revealed conservative of their short-genome electropherotypes and double-stranded RNA migration patterns characteristics of group B rotaviruses. To our knowledge, our previous study (L.J. Saif, K.V. Brock, D.R. Redman, and E.M. Kohler, Vet. Rec. 128:447-449, 1991) and this report are the first description of bovine group B rotaviruses (in a mixed infection with bovine coronavirus or singly in fecal contents) in adult cows with diarrhea and this is the first report of short genome electropherotypes among group B rotaviruses. PMID- 8727927 TI - Molecular typing of Borrelia burgdorferi from Lyme disease patients by PCR restriction fragment length polymorphism analysis. AB - Ninety-three Borrelia burgdorferi isolates obtained from erythema migrans lesions or blood of Lyme disease patients in Westchester County, N.Y., between 1991 and 1994 were characterized by PCR-restriction fragment length polymorphism (PCR RFLP) analysis of the 16S-23S rRNA gene spacer. All isolates could be classified into three distinct RFLP types. Among the 82 skin biopsy isolates studied, 21 (25.6%) were type 1, 37 (45.1%) were type 2, and 21 (25.6%) were type 3. Three (3.7%) cultures contained a mixture of two isolates with distinct RFLP types. The 11 isolates cultured from blood showed a similar predominance of RFLP type 2 (6 of 11; 54.5%) relative to types 1 (2 of 11; 18.2%) and 3 (3 of 11; 27.3%). For one patient both skin and blood isolates were cultured, and RFLP analysis revealed that these isolates differed from one another. This study demonstrates that there is genotypic heterogeneity in B. burgdorferi strains infecting Lyme disease patients, and this typing approach may allow differentiation of isolates with various degrees of pathogenic potential. PMID- 8727928 TI - Comparison of urinary bladder and ear biopsy samples for determining prevalence of Borrelia burgdorferi in rodents in central Europe. AB - PCR was used to compare urinary bladder and ear biopsy samples from four European species of wild rodents for the presence of Borrelia burgdorferi sensu lato. From 60 paired comparisons of bladder and ear biopsy samples, the PCR results were concordantly positive or negative in 43 samples (71.7%). Of the 17 which differed, 14 bladder samples were positive and ear samples were negative while the converse occurred for three samples. Thus ear biopsy samples led to a significantly lower estimate of infection than bladder biopsy samples. This suggests that the use of ear biopsy samples in epidemiological studies of B. burgdorferi in Central European rodents is likely to lead to underestimates of the prevalence. PMID- 8727929 TI - Detection of parvovirus B19 DNA in bone marrow cells by chemiluminescence in situ hybridization. AB - A chemiluminescence in situ hybridization method was developed for the search of B19 parvovirus DNA in bone marrow cells, employing digoxigenin-labeled B19 DNA probes, immunoenzymatically detected with a highly sensitive 1,2-dioxetane phosphate as chemiluminescent substrate. The light emitted from the in situ hybridized probe was analyzed and measured by a high-performance luminograph connected to an optical microscope and to a personal computer for the quantification of the photon fluxes from the single cells and for image analysis. The chemiluminescence in situ hybridization was applied to bone marrow cell smears of patients with aplastic crisis or hypoplastic anemia, who had been previously tested by in situ hybridization with colorimetric detection, dot blot hybridization, and nested PCR. The chemiluminescent assay provided an objective estimation of the data, proved specific, and showed an increased sensitivity in detecting B19 DNA compared with in situ hybridization with colorimetric detection. PMID- 8727930 TI - Direct identification of Mycobacterium species in Bactec 7H12B medium by gas liquid chromatography. AB - Twenty-nine Mycobacterium reference strains representing 10 species and 60 mycobacterial cultures isolated from sputum specimens were studied. These cultures were grown in Bactec 7H12B medium (Becton Dickinson and Co., Paramus, N.J.) supplemented with oleic acid-albumin-dextrose-catalase enrichment broth (Becton Dickinson and Co., Cockeysville, Md.). The cultures were analyzed by gas liquid chromatography for their fatty acids, secondary alcohols, and mycolic acid cleavage products. All of the clinical isolates could be identified by comparing their gas-liquid chromatography profiles with those of the reference strains. The data indicate that this method significantly shortens the turnaround time and could be used for the early detection and identification of mycobacterial species. PMID- 8727931 TI - Stability of amoxicillin-clavulanate in BACTEC medium determined by high performance liquid chromatography and bioassay. AB - The stabilities of amoxicillin (16 micrograms/ml) and clavulanate (8 micrograms/ml), alone and in combination in BACTEC medium (Middlebrook 7H12B medium), were determined by high-performance liquid chromatography (HPLC) and bioassay. By HPLC, the half-life of amoxicillin (trihydrate and sodium) in combination with clavulanate in nonradiolabelled 7H12B medium was 6.7 days, whereas the half-life of clavulanate in combination with amoxicillin was 2.0 days. By bioassay, the half-lives of amoxicillin trihydrate and clavulanate in radiolabelled 7H12B medium were comparable (7 and 2 days, respectively) to those determined by HPLC. When clavulanate was tested alone, the half-life was determined to be 1.88 days by HPLC and 1.87 days by bioassay. The relatively short half-life of clavulanate can be adjusted by a procedure of "topping up," or adding one-half the concentration of clavulanate every second day, in order to allow accurate amoxicillin-clavulanate MIC testing with the BACTEC mycobacterial susceptibility system. PMID- 8727932 TI - New drug susceptibility test for Mycobacterium tuberculosis using the hybridization protection assay. AB - We developed a novel method for early detection of drug-resistant strains of Mycobacterium tuberculosis by using the hybridization protection assay (HPA). The number of viable bacteria during the incubation period correlated well with the number of relative light units measured by the HPA. In addition, the relative light unit values of susceptible strains on the first, third and fifth days of incubation were significantly different from those of resistant strains for both isoniazid and rifampin. Our results suggest that after isolation of the organism from clinical specimens, drug-resistant strains of M. tuberculosis are accurately detected by the HPA even after 1 day of incubation with the drug. PMID- 8727933 TI - Comparison of BacT/Alert FAN medium with BACTEC NR660 Plus 26A medium. AB - A total of 6,010 blood culture sets were obtained from adult patients with suspected bacteremia or fungemia. The overall recovery of organisms was equivalent in two systems used, BacT/Alert FAN medium and BACTEC NR660 Plus 26A medium. PMID- 8727934 TI - Necrotizing cerebritis in an allogeneic bone marrow transplant recipient due to Cladophialophora bantiana. AB - We describe a necrotizing cerebritis in an allogeneic bone marrow transplant recipient caused by the neurotropic, dematiaceous fungus Cladophialophora bantiana. The patient presented 7 months after bone marrow transplantation with fever and sudden onset of left-sided weakness, followed shortly by cranial nerve III and VI palsies. The patient had a lesion (3.0 by 2.0 by 2.0 cm) of the right midbrain with extension to the pons, the left brain stem, and the right superior and the middle cerebellar peduncles. The diagnosis was made by microscopic examination and culture of a brain biopsy. PMID- 8727935 TI - Fatal, disseminated Acremonium strictum infection in a neutropenic host. AB - Disseminated Acremonium strictum infection in a neutropenic patient is reported. Positive fecal cultures preceded positive cutaneous and blood cultures by 18 and 21 days, respectively, which suggests gastrointestinal colonization and invasion as initiating events. Microscopic examination of cutaneous biopsy and pulmonary specimens revealed hyphae, phialides, and phialoconidia in vivo. These adventitious forms also can occur in infections due to other phialidic fungi such as Fusarium and Paecilomyces species and can be misdiagnosed as Candida species. Budding cells also can occur in vivo for species of Fusarium, Paecilomyces, and apparently Acremonium, further adding to the potential for misdiagnosis. The occurrence of adventitious forms in infections caused by species of Acremonium, Fusarium, Paecilomyces, Scedosporium, and Blastoschizomyces is suggested as a mechanism for dissemination of infection and as an explanation of the relatively higher frequency of positive blood cultures in these cases. PMID- 8727936 TI - Evaluation of CMV Brite kit for detection of cytomegalovirus pp65 antigenemia in peripheral blood leukocytes by immunofluorescence. AB - The CMV Brite antigenemia kit was compared with culture and an established cytomegalovirus pp65 antigenemia assay (CMV AG). Of 300 clinical specimens tested, 92 were positive by CMV Brite, 83 were positive by CMV AG, and 34 were positive by culture. Discrepancies could be attributed to anticytomegalovirus therapy or low-level antigenemia. PMID- 8727937 TI - Demonstration of a lack of synergistic effect of rotavirus with other diarrheal pathogens on severity of diarrhea in children. AB - The severity of group A rotavirus (RV) diarrhea was compared with that of mixed infections of RV with diarrheagenic Escherichia coli, Vibrio cholerae O1, and Shigella species by a scoring system. The severity of mixed infections of RV and E. coli was the same as that of infections with RV alone. RV infections mixed with V. cholerae and Shigella species mimicked cholera and shigellosis, respectively. PMID- 8727938 TI - Improved detection of JC virus DNA in cerebrospinal fluid for diagnosis of AIDS related progressive multifocal leukoencephalopathy. AB - Several methods to increase the sensitivity of JC virus (JCV) DNA detection in cerebrospinal fluid (CSF) for a noninvasive diagnosis of AIDS-related progressive multifocal leukoencephalopathy (PML) were investigated. When CSF collected at clinical presentation was tested, JCV DNA was detected in 8 of 19 patients with PML by standard PCR (sensitivity, 42%; 95% confidence interval [CI], 21 to 66%) and in 14 of 19 by nested PCR (sensitivity, 74% [95% CI, 49 to 90%]; P = 0.014 [McNemar's test]. For multiple serial CSF samples, standard PCR yielded JCV DNA for 11 of 19 PML patients (sensitivity, 58% [95% CI, 34 to 79%]) and nested PCR yielded JCV DNA for 17 of 19 patients (sensitivity, 90% [95% CI, 66 to 98%]; P = 0.014). The majority of the false-negative samples were found to contain PCR inhibitors. Standard PCR did not detect JCV DNA in CSF from any of the 83 AIDS patients with other diagnosis (100% specificity [95% CI, 95 to 100%]); JCV DNA was found in CSF from one control patient by nested PCR (99% specificity [95% CI, 93 to 100%]). PMID- 8727939 TI - PCR for diagnosis and follow-up of two cases of disseminated toxoplasmosis after kidney grafting. PMID- 8727940 TI - IS6110-based PCR methods for detection of Mycobacterium tuberculosis. PMID- 8727941 TI - Chronological changes in the complement system in sepsis. AB - The time courses of serum complement levels and the severity of sepsis were compared in two groups of septic patients, one in which the patients survived (surviving group) and one in which they did not (nonsurviving group). The components of the complement system, namely, C3a, C4a, C5a, CH50, C3, C4, and C5, were measured at several points in time after the diagnosis of sepsis had been established. A 2-antibody radioimmunoassay was used to measure C3a, C4a, and C5a; the latex agglutination test was used to measure C3 and C4; nephelometry was used to measure C5; and Meyer's 50% hemolysis method was used to measure CH50. Following the diagnosis of sepsis, the levels of CH50, C3, and C4 were significantly lower in the nonsurviving than the surviving group, while the levels of C3a and C4a were significantly higher in the nonsurviving than the surviving group. The C5a levels were significantly higher in the nonsurviving than the surviving group, although no significant intergroup differences were subsequently noted. These results suggest that the serum levels of C3a, C4a, C5a, CH50, C3, and C4 could serve as indices of the severity of sepsis. Thus, monitoring the complement system may be useful for predicting the outcome of patients with sepsis. PMID- 8727942 TI - Immunohistochemical analysis of p53 and ras p21 expression in colorectal adenomas and early carcinomas. AB - To further investigate whether multiple genetic changes are involved in the development of colorectal cancer, we performed an immunohistochemical analysis of p53 and ras p21 protein expression in 139 specimens of colorectal adenoma with varying degrees of dysplasia, 57 specimens of early cancer with an adenomatous component, and 12 specimens of superficial early cancer without any adenomatous component. Positive p53 staining was found in 15% of the adenomas with moderate dysplasia and in 42% of the adenomas with severe dysplasia or intramucosal carcinoma (IMCA). Positive immunostaining of p53 was observed to be significantly correlated with the degree of dysplasia and the depth of invasion, as was the expression of ras p21. However, a closer correlation was observed with the increasing size of the adenomas. Furthermore, p53 staining was positive in 42% of the 12 superficial early cancer specimens, while ras staining was positive in only 1 specimen (8%). These results indicate that p53 gene overexpression may play some biological role in both the adenoma-to-carcinoma sequence and in de novo cancer development, whereas ras p21 expression may not be as involved in de novo cancer development as in the malignant conversion of colorectal adenomas. PMID- 8727943 TI - Plasma hepatocyte growth factor levels are increased in systemic inflammatory response syndrome. AB - Interleukin-1 (IL-1), a cytokine released from macrophages by endotoxin stimulation, has been shown to upregulate the genetic expression of the hepatocyte growth factor (HGF). The present study was conducted to determine whether plasma HGF is increased in patients with systemic inflammatory response syndrome (SIRS). The plasma levels of HGF, endotoxin, and beta-glucan were measured in 41 surgical patients without hepatic diseases, 18 of whom had been diagnosed with sepsis, and 33, with nonseptic SIRS. The plasma HGF was found to be significantly increased in the 18 patients with sepsis, at 0.69 +/- 0.47 ng/ml (mean +/- SD), and in the 23 patients with nonseptic SIRS, at 0.49 +/- 0.37 ng/ml, compared to values in 40 normal controls, at 0.10 +/- 0.03 ng/ml (P < 0.001). No significant correlations were observed between the plasma levels of HGF and endotoxin (r = 0.02) or beta-glucan (r = -0.05) in any of the patients; however, plasma HGF was significantly correlated with the WBC count (r = 0.34, P < 0.05) and with total bilirubin (r = 0.45, P < 0.01). Plasma HGF was also strongly correlated with alanine transaminase (ALT) in 8 patients with ALT levels higher than 50 U/l (r = 0.70), but there was no such correlation in 33 patients with ALT levels of 50 U/l or less (r = 0.30). Thus, although the clinicopathologic significance of HGF is not well understood, the present findings indicate that plasma HGF increases in response to infection or inflammation. PMID- 8727944 TI - Urinary thromboxane B2 as an indicator of acute rejection in human liver transplantation. AB - Urinary thromboxane B2 (u-TXB2) was measured and analyzed after a human liver transplantation in 28 patients (30 transplantations) who underwent an orthotopic liver transplantation. Our results showed that the u-TXB2 levels exceeded 3.0 micrograms/mmol creatinine in only 2 of the 13 cases that had a favorable postoperative course. In 10 of the 11 episodes of acute rejection, the u-TXB2 levels exceeded 3.0 micrograms/mmol creatinine. In 6 episodes of acute rejection, the TXB2 levels were more than 5.0. In 4 out of 6 episodes of infection unassociated with rejection, the u-TXB2 values were between 3.0 and 4.9 micrograms/mmol creatinine. In 2 episodes of liver necrosis the TXB2 value reached 5.3 in one and 0.9 in the other. In conclusion, the u-TXB2 level was observed to be elevated in cases of acute rejection, infection, or necrosis. The diagnosis of acute rejection on the basis of u-TXB2 showed a sensitivity of 58.8%, a specificity of 93.3%, and an accuracy of 75.0% for a threshold level of 3.0 micrograms/mmol creatinine, and a sensitivity of 85.7%, a specificity of 79.2%, and an accuracy of 80.6% for a threshold level of TXB2 of 5.0 micrograms/mmol creatinine. These results indicate that the serial determination of u-TXB2 is a useful diagnostic means for predicting acute rejection after liver transplantation. PMID- 8727946 TI - Inflammatory reactions after vascular prosthesis implantation: a comparison of gelatin-sealed and unsealed Dacron prostheses. AB - Despite widespread use of the gelatin-sealed knitted Dacron prosthesis (GDP) in clinical practice owing to its zero porosity, the biological impacts of this graft are still controversial. We conducted a randomized controlled study on 50 patients undergoing abdominal aortic aneurysm repair to evaluate the inflammatory reaction to GDP (n = 25) and unsealed knitted Dacron prostheses (UDP, n = 25). There were no significant differences in the mean age, size of the aneurysm, operative time, blood loss, or transfusion requirements between the GDP and UDP groups. During the first 7 postoperative days (PODs), slight fever and leukocytosis were noticed in both groups. Significant differences in maximum body temperature, leukocyte count, and plasma C-reactive protein concentration were observed between the GDP and UDP groups on POD 14:37.2 +/- 0.5 degrees C vs 36.9 +/- 0.3 degrees C (P = 0.019), 8,151 +/- 1,788/microliters vs 6,914 +/- 1,501/microliters (P = 0.015), and 32.6 +/- 27.5 mg/l vs 19.0 +/- 15.8 mg/l (P = 0.048), respectively. By POD 21, however, there were no detectable differences in these variables. Thus, we concluded that GDP caused an inflammatory reaction in the 2nd week after implantation, but ultimately there were no significant differences from UDP by the 3rd week. PMID- 8727945 TI - Serum tumor marker kinetics and the clinical course of patients with advanced breast cancer. AB - Serum carcinoembryonic antigens (CEA), CA 15-3, and tissue polypeptide antigens (TPA) have been used in monitoring the clinical course of patients with breast cancer. However, recent reports have suggested that the serial levels of these markers during therapy do not always correlate with the response to therapy. To clarify the usefulness of the serial combination assay of these markers in monitoring the clinical course of patients during therapy, we investigated the relationship between the initial changes and the kinetic patterns of the markers after therapy and the objective responses. When an increase or decrease of over 20% in these markers is taken to be significant, then the initial changes in all three markers significantly correlated with the therapeutic responses (P < 0.01). Five distinct kinetic patterns in the marker levels were observed. A paradoxical kinetic pattern of CEA and CA 15-3 levels--that is, an "initial surge and subsequent drop"--was seen in one-third of the responders. The TPA levels tended to exhibit a "steady decline" pattern in those responders. The sensitivity and specificity of the kinetic patterns to predict the clinical courses were significantly higher than those obtained from the analysis of initial changes. These findings thus suggest that adequate knowledge of the unique kinetics of each marker may help to make a more accurate prediction of the therapeutic responses. PMID- 8727947 TI - Early results of a reinforced biosynthetic ovine collagen vascular prosthesis for small arterial reconstruction. AB - The efficacy of a reinforced biosynthetic ovine collagen (RBOC) vascular prosthesis developed for small arterial reconstruction was assessed by examining 30 grafts in 29 patients with arteriosclerosis obliterans. The operative procedures performed were femorofemoral bypass in 2 patients, above-knee femoropopliteal bypass in 28 patients, and below-knee femoropopliteal bypass in 1 patient. Femoropopliteal bypass was simultaneously performed in two patients undergoing femorofemoral bypass using one or two grafts. The indications for surgery were intermittent claudication in 27 patients and to salvage the limb in 2 patients. The longest follow-up period was 49 months, and there were six graft failures, occurring 1, 1, 9, 17, 17, and 23 months after implantation, respectively; caused by compression of the graft from outside in two, infection in one, anastomotic intimal hyperplasia in one, and unknown factors in two. Thus, the primary cumulative patency rate for above-knee femoropopliteal bypass at 3 years was 83.7%, and the secondary patency rate was 91.2%. No aneurysmal change was observed. Moreover, the RBOC was able to be used without preclotting, and its handling and suturing characteristics were satisfactory. Our findings suggest that this vascular prosthesis may be an acceptable alternative for above-knee femoropopliteal bypass. PMID- 8727948 TI - Serial growth of human malignant fibrous histiocytoma xenografts in immunodeficient mice. AB - Malignant fibrous histiocytoma (MFH) is on of the most common soft tissue sarcomas of adulthood, the only treatment for which involves surgical resection of the extremities and retroperitoneum, while no standard postoperative adjuvant chemotherapy has been established. We report herein on the establishment of a serially transplantable MFH strain in immunodeficient mice. An intraperitoneal tumor was resected from a patient with multiple recurrent MFH, inoculated into the subcutaneous tissue of mice with severe combined immunodeficiency (SCID), and established as a serially transplantable MFH strain, MH-1. The chemosensitivity of MH-1 was similar to that of the original fresh surgical specimen, as confirmed by the 3-(4,5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H tetrazolium bromide (MTT) test. We believe that this serially transplantable strain will be useful for further studies on chemotherapy effective against MFH. PMID- 8727949 TI - Role of insulin resistance in decreasing lipoprotein lipase activity in tumor bearing rats. AB - The role of insulin resistance in the tumor-induced decrease in tissue lipoprotein lipase (LPL) activity was studied in vivo and vitro in methylcholanthrene-induced sarcoma-bearing rats. Intraperitoneal (i.p.) injection of 2U of regular insulin resulted in high-adipose LPL activity in control rats (CTR) of 122.0 +/- 42.4 U/mg tissue, but it had little effect on tumor-bearing rats (TBR), which showed a value of only 9.6 +/- 5.5 U/mg tissue (P = 0.002). When adjusted for serum insulin concentrations, adipose LPL activity remained significantly different between the TBR and CTR at 0.19 +/- 0.17 and 0.78 +/- 0.29 U/mg tissue, respectively (P = 0.02). Following the in vitro incubation with either 1.44 g/l glucose of 1 x 10-8 U insulin of adipose tissue fragments obtained from the TBR and the CTR, measurable LPL activity was maintained in the tissue from the CTR for 2 h but not in that from the TBR. These results suggest that the decreased LPL activities seen in the tumor-bearing state may be mediated by insulin resistance. PMID- 8727950 TI - Proliferation of osteoclast-like giant cells in a metastatic bone tumor from stomach cancer: report of a case and analysis of the autopsy findings. AB - A 70-year-old man who had undergone esophagectomy with reconstructive surgery using a portion of the stomach 5 years earlier for esophageal cancer was admitted to our hospital after a routine endoscopy and histological examination of a biopsy specimen revealed poorly differentiated adenocarcinoma in the stomach. A gastrectomy and intrathoracic esophagojejunostomy was performed on January 20, 1993; however, the patient suffered a cerebral infarction and died of septic shock on April 9, 1993. At autopsy, metastatic tumors were macroscopically observed in various organs, including a bone tumor measuring 1.0 cm in diameter in the L4 vertebra. To clarify the origin of the bone tumor, we conducted histological and immunohistochemical examinations. Histological examination revealed a mixture of osteoclast-like giant cells (OGCs) and poorly differentiated adenocarcinoma cells, although no histologic features of OGCs were observed either in a primary site or in any of the multiple metastatic lesions. On immunohistochemistry, adenocarcinoma cells in the bone stained positively for the carcinoembryonic antigen (CEA), whereas no staining for CEA was observed in the OGCs which demonstrated negative staining for all the antigens of epithelial markers. These findings led us to conclude that this bone tumor had metastasized from the stomach cancer and that the OGCs may have originated from mesenchymal cells reacting to the adenocarcinoma cells. PMID- 8727951 TI - Choledochocele: changing trends in diagnosis and management. AB - Eighty-four patients with choledochocele collected from the world literature and one personal observation are reviewed. The main issues regarding clinical presentation, diagnostic work-up, and the treatment of this uncommon lesion are discussed. Abdominal pain was the most common clinical feature (91% of cases), followed by pancreatitis (38%), nausea or vomiting (35%), and jaundice (26%). In addition, associated lithiasis was found in 43% of the cases. Endoscopic retrograde cholangiopancreatography was the most useful diagnostic procedure and resulted in a correct diagnosis in all but one of the patients investigated by this method. Surgical excision of the duodenal luminal portion of the choledochocele was the treatment most commonly used (65% of cases). In recent years, operative endoscopy has also been increasingly used, with good results. PMID- 8727952 TI - Successful utilization of the median sternotomy approach in the management of descending necrotizing mediastinitis: report of a case. AB - We describe herein the case of a patient in whom a median sternotomy was successfully employed for mediastinal drainage in the treatment of descending necrotizing mediastinitis (DNM). Although most reports describe cervical or thoracotomy approaches, our experience strongly suggests that median sternotomy is a satisfactory alternative approach for treatment of this disease. PMID- 8727953 TI - Aggressive angiomyxoma occurring in the scrotum: report of a case. AB - The authors recently treated a case of aggressive angiomyxoma occurring in the left scrotum. A 61-year-old Japanese man was diagnosed as having an inguinal irreducible hernia that descended to the bottom of the scrotum preoperatively. However, during the operation a large tumor was found between the spermatic fascia and the skin pressing the testis upward. The tumor measured 20 x 15 x 15 cm, was yellowish with a smooth surface encapsulated by a membrane similar to the pleura, and the cut surface was gelatinous. The histological findings indicated that the tumor had thick-walled vessels which had prominently increased in size, delicately waved collagen fibers, and stromal cells in a myxoid background. The patient has remained recurrence-free as of 11 months after surgery. PMID- 8727954 TI - Development of postoperative delirium in relation to a room change in the general surgical unit. AB - To analyze the relationship between the development of postoperative delirium and a change of the patient's room, 1,006 cases of patients who had undergone surgery with general anesthesia were reviewed. Postoperative delirium developed in 84 (8.3%) cases. On the basis of symptomatic features, postoperative delirium was divided into four types: (1) excitement type, (2) excitement-hallucination type, (3) hallucination type, and (4) disorientation type. Of the 31 excitement-type cases, 21 developed within the 2nd postoperative day (POD) while 27 of 29 hallucination types developed after POD 2. Of 29 hallucination types, 22 developed after a room change while 20 of these 22 cases were transferred to a single room before POD 2. A quiet, dark, and isolated environment in a single room is suggest to contribute to the development of hallucinations. The development of postoperative delirium with hallucinations alone should thus be taken into consideration whenever a room change is decided. PMID- 8727955 TI - A novel paper cuff for vascular reconstruction in canine liver transplantation. AB - A new, simple method of producing optimal cuffs using thin paper and then setting it in resin for vascular reconstruction in canine liver transplantation is herein described. Thin Paper was cut into a paper tape strip. By fixing both ends of this paper tape, a cylinder paper core of any desired size could thus be obtained. The paper core was immersed into a two-liquid mixture-type resin, removed, and left to harden. The paper cuffs (0.2 mm or less in thickness) were then used for anastomoses of the portal vein and the infrahepatic inferior vena cava in a series of 10 consecutive canine liver transplantations. Out of 10 animals, 8 survived for more than 5 days. The vascular patency in these 8 animals at the cuff anastomotic sites was 100% at postmortem. We therefore conclude that this paper cuff appears to be useful for various types of experimental liver transplantations in large animals. PMID- 8727956 TI - The role of membranes in lens ageing and cataract. Proceedings of a meeting. Naples, March 13-14, 1995. PMID- 8727957 TI - Transgenic mice: models for the study of cataractogenesis. A minireview. AB - Genetic manipulation followed by (over)expression of the transgene product in lens tissue results in: (I) inhibition of denucleation of lens fibers; (II) interference with lens cell differentiation; (III) cataract formation. The events reviewed are preceded by membrane damage, perturbed assembly of the IF cytoskeleton and distortion of the cytoskeleton-membrane complex. PMID- 8727958 TI - Cataract mutations as a tool for developmental geneticists. AB - Lens development as a multistep process can be analyzed by the investigation of distinct cataract mutants. Since the mutant genes are molecularly characterized, the function of the wild-type allele can be deduced. Besides some mutations affecting the lens induction, which are not yet characterized at the molecular level, mainly mutations affecting the crystallin genes are discussed. In particular, for the murine gamma-crystallin genes 8 different mutations are described in the mouse, which lead to different, distinguishable phenotypes. The distinct and complex phenotypes cannot be explained solely by the changed physiochemical properties of the altered crystallin packaging, but point to a regulatory function of the crystallins during lenticular development and differentiation. PMID- 8727959 TI - Prevention of the fructation-induced inactivation of glutathione reductase by bovine alpha-crystallin acting as a molecular chaperone. AB - We had previously shown that glutathione reductase was inactivated by glycation, and decided to use this model system to see whether bovine alpha-crystallin could provide protection, and whether its modification had an effect. Other proteins were used as a comparison. Both forms of alpha-crystallin prevented loss of enzyme activity on incubation with fructose. No such protection was profferred by the other proteins, suggesting bovine alpha-crystallin acted in a chaperone-like manner. PMID- 8727960 TI - Calf lens alpha-crystallin, a molecular chaperone, builds stable complexes with beta s- and gamma-crystallins. AB - Calf water-soluble (WS) crystallins consist of high-molecular weight (HM), alpha , beta H-, beta L-, beta s- and gamma-crystallins. alpha-Crystallin as a molecular chaperone forms a structure with a central hole, known as Carver's model. The only crystallins fitting into this central cavity are beta s- and gamma-crystallins, with native molecular weights of 28 and 18.5-21 kD, respectively. The beta s-crystallin is loosely bound to this structure, whereas with the application of 7M urea in the sample, beta s-, may be some beta L components and all gamma-crystallins emerge from the central hole. Although WS and water-insoluble (WI) crystallins display the same immunologic determinants, there is an appreciable difference in electrophoretic mobility already in the young calf lens. alpha-Crystallins from the WI part demonstrate a clear cathodic shift. WI beta- and gamma-crystallins show smaller but well perceptible cathodic shifts. The chaperone function of alpha-crystallin preserves the original structures of beta s- and gamma-crystallins for aggregation and other influences. PMID- 8727961 TI - The influence of some post-translational modifications on the chaperone-like activity of alpha-crystallin. AB - We investigated the influence of phosphorylation, glycation, carbamylation and oxidative modification on the capacity of alpha-crystallin to protect beta crystallins against heat denaturation. Simple modification of lysine residues by early glycation or carbamylation had no effect. However, late (cross-linking) glycation products and oxidative modifications decreased the chaperone-like activity of alpha-crystallin. Homopolymers of alpha A-crystallin had a higher protecting capacity compared with those of alpha B-crystallin. The in vivo phosphorylated forms of especially alpha A- but also alpha B-crystallin revealed a somewhat better protecting ability than the respective non-phosphorylated forms. PMID- 8727962 TI - alpha-Crystallin: molecular chaperone and heat shock protein. AB - The relationship of alpha-crystallin with the family of small heat shock proteins has led to the discovery that the basic subunit alpha B-crystallin can, like other heat shock proteins, protect cells against heat stress. Here we show that the acidic subunit alpha A-crystallin, which in contrast to alpha B-crystallin is expressed mainly in the eye lens, shares this property. Furthermore we have investigated the in vitro molecular chaperone-like behavior of the natural mutant alpha A ins-crystallin that has a large insert peptide and occurs in rodents. We have found the chaperone-like activity of the mutant to be diminished compared to that of the wild type alpha A-crystallin. PMID- 8727963 TI - Calcium binding properties of beta-crystallins. AB - beta-Crystallins, the oligomeric proteins of the eye lens, were found to bind calcium ions with low affinity, in the millimolar range. Calcium was found to induce conformational changes in the secondary and tertiary structure of beta crystallins. While these changes in conformation are seen in low ionic strength media, they are masked when the protein is dissolved in an intermediate ionic strength medium, where oligomers of lower molecular weight are formed. The calcium binding takes place in these conditions. The behaviour of this protein with respect to calcium binding is reviewed. PMID- 8727964 TI - Calcium-induced disruption of the lens cytoskeleton. AB - Ionic homeostasis is essential to lens clarity and the lens epithelium plays a large part in homeostasis, through vectorial transport. In most epithelia maintenance of vectorial function depends on the cytoskeleton. Capsule/epithelium preparations from human donor lenses have been used for immunohistochemical investigations of both normal epithelial cell cytoskeletal structure, and of structural changes induced by increasing cell calcium. A sustained increase in intracellular calcium, induced by incubation with A23187 and W7, led to a loss of cytoskeletal organisation. Changes induced depolymerisation of cytoplasmic actin filaments, disaggregation of microtubules and initial thickening, followed by breakdown, of vimentin intermediate filaments. In addition, spectrin staining, normally confined to the basal-lateral plasma membranes, became diffuse within the cytoplasm. Such disruptions of cytoskeletal structure would decrease ionic control, leading to increasing osmotic stress. PMID- 8727965 TI - Calpains in the human lens: relations to membranes and possible role in cataract formation. AB - Calpains are Ca-activated neutral proteases present in all cells together with an endogenous inhibitor, calpastatin. Proposed substrates are; cytoskeletal proteins like microtubules and actin, protein kinases such as PKC and membrane-bound enzymes like Ca-ATPase and the Ca-channel. In lenses from different species calpains have been detected in decreasing amounts from the epithelium to the cortex to the nucleus. Several substrates for calpain in the lens have been demonstrated: crystallins, vimentin, actin, beaded filaments and MP26 among others. Both studies on animal models and capsulorhexis indicate that calpains are mainly involved in cortical cataract. PMID- 8727966 TI - Chicken CP49: significant or paltry? An evolutionary perspective. AB - The lens-specific intermediate filament (IF) proteins CP49 and filensin have been identified in a large number of evolutionary divergent species. In the chick lens both CP49 and filensin have been identified. Recently a unique variant of CP49, CP49ins has been identified. CP49ins contains an insertion of 49 amino acids in helix 1b giving it characteristics similar to the lamin-like cytoplasmic IF of invertebrates. The presence of this variant form in the chick lens poses some important questions about the co-assembly of the two CP49 forms and also the evolutionary relationship with invertebrate IF proteins. PMID- 8727967 TI - The intermediate filament cytoskeleton of the lens: an ever changing network through development and differentiation. A minireview. AB - In the eye lens, intermediate filament proteins form two morphologically distinct polymers, 10-nm intermediate-sized filaments and beaded filaments. Coincidently, the beaded filament polymer and the proteins filensin and CP49 are specific to lens fibre cells and are therefore excellent markers for fibre cell differentiation. In the adult lens, filensin and CP49 are maintained throughout all stages of lens fibre cell differentiation whilst vimentin is apparently lost at a specific stage from the deeper cortical fibres. The expression of CP49 and filensin is coincident with the presence of beaded filaments suggesting these proteins are filament components. In association with alpha-crystallin, CP49 and filensin form beaded filaments in vitro. During fibre cell differentiation, filensin and CP49 are post-translationally modified. In the case of filensin, proteolysis results in two functionally distinct fragment sets, one derived from the alpha-helical rod domain and the other from the C-terminal tail domain of filensin. It is proposed that both filensin and CP49 are critically involved in organising the cytoplasmic and plasma membrane domains of the fibre cell and therefore essential to the optical properties of the lens. PMID- 8727968 TI - Interaction of glycyl-phenylalanine with carbohydrates as a model of N-terminal glycation of proteins. AB - Glycation is thought to be a factor in the development of cataract in elderly and diabetic patients. In this report, we describe our initial investigations on the reaction of the dipeptide glycyl-phenylalanine (GF) with different carbonyl compounds as a model of protein glycation. The results obtained demonstrate that the interaction of GF with carbohydrates has features in common with glycation of lens membrane proteins and lens crystallins. PMID- 8727969 TI - Lens proteins changes induced by sugars and pyridoxal phosphate. AB - Glucose, galactose and pyridoxal phosphate (PLP) bind to lens protein amino groups causing changes in absorbance and fluorescence spectra and inducing aggregation. Sugars and PLP simultaneously cause an increase in fluorophore and chromophore formation, but a decreased aggregation, compared to PLP alone. PLP binds to lens protein amino groups decreasing the sugar binding, but in preventing glycation by PLP attention should be paid to the consequences of its own binding to proteins in diabetes. PMID- 8727970 TI - Main intrinsic polypeptide and water movement--recent developments and future prospects. AB - Age-related lens opacity is the major cause of loss of vision affecting more than half of the world's blind. The development of cataract is associated with changes in the structure of the lens. The lens consists largely of closely packed fibre cells forming a transparent syncytium. The main intrinsic polypeptide (MIP) of the lens fibre cells forms 40% of the fibre membrane, and forms a system of membrane channels. The precise structure and function of these channels remains obscure, although their role as large water-filled channels has been suggested in maintenance of structural integrity as well as their role in transport of nutrients. MIP is a member of a large family of channels of common ancestry. Some of the most recent studies of structure and function of this family have been carried out on CHIP (a water channel found in red blood cells) which is a member of this family closely related to MIP. Both CHIP and MIP m-RNA's have been expressed in oocyte membranes, and water permeabilities per channel have been measured. Surprisingly, CHIP and MIP channels have high and low water permeabilities, respectively. This is discussed in terms of recent high resolution structure determinations, the lens fibre MIP function, and future studies. PMID- 8727972 TI - Epithelial cell-fibre coupling in the equator of the human lens. AB - A detailed freeze fracture study of human lenses of various ages revealed that coupling between epithelial cells and fibres occurs only in a limited area in the equator of the lens; it also demonstrated that the extent of coupling is independent of age. The developing fibres remain coupled to their original neighbours and no other contacts between epithelial cells and fibres appear to be established. PMID- 8727971 TI - Differential expression of sodium pump catalytic subunits in the lens epithelium and fibers. AB - Na,K-ATPase in lens epithelium plays a key role in conducting sodium-potassium transport. The purpose of this study was to test whether epithelium or fiber cells can synthesize new Na,K-ATPase protein in response to an increase of membrane permeability. Western blot methodology was used to identify Na,K-ATPase alpha subunit polypeptides in membrane material isolated from lens cells. As judged by immunoblot density, epithelial cell membrane material isolated from porcine lenses cultured 24 h with 1 microM amphotericin B contained more Na,K ATPase alpha subunit polypeptide than epithelial material isolated from control lenses. This increase stemmed from the apparent synthesis of Na,K-ATPase alpha 2 isoform polypeptide by the epithelium; Na,K-ATPase alpha 1 isoform polypeptide abundance was not detectably altered. The apparent amphotericin B-induced expression of Na,K-ATPase alpha 2 was seen in lens epithelial cells but not fiber cells. This study suggests that the epithelium of the adult porcine lens may be capable of expressing additional sodium pump molecules of the alpha 2-subtype when membrane permeability is increased. PMID- 8727973 TI - Thermodynamic phase transition parameters of human lens dihydrosphingomyelin. AB - Dihydrosphingomyelin (DHS) is the major phospholipid in the human lens. The influence of this phospholipid on membrane structure and function is not known. In this study we used infrared spectroscopy to determine the thermodynamic and molecular structural properties of the hydrocarbon chains of DHS membranes isolated from human lenses. The phase transition temperature of human lens DHS was 9 degrees C higher than for bovine brain sphingomyelin membranes and 14 and 7 degrees C higher than human lens cortical and nuclear membranes, respectively. This increase in the phase transition temperature results in 20% increase in lipid order at 36 degrees C in comparison to that of native membranes and bovine brain sphingomyelin. DHS is likely to provide structural order to the hydrocarbon chain region and upholds the integrity of native membranes under oxidative conditions. PMID- 8727974 TI - Aging affects the conformation of cholesterol in the human eye lens. AB - The distribution of 3-beta-OH-cholesterol along the optical axis of 20 human eye lenses of different ages was measured using confocal Raman microspectroscopy in combination with the 3-beta-hydroxysteroid-specific probe filipin. This non destructive technique allow a spatially very detailed study of the distribution of 3-beta-OH-cholesterol in individual lenses. It revealed age-related changes of the 3-beta-OH-cholesterol distribution along the optical axis. The 3-beta-OH cholesterol was found to be distributed asymmetrically along the optical axis and this distribution was found to change with age. The relative 3-beta-OH cholesterol content in the anterior part of the lens decreases significantly with increasing age compared with its equal posterior counterpart. Additionally all the lenses have a very low 3-beta-OH-cholesterol content in the central part of their nuclear region, the area where the primary lens fibres are located. PMID- 8727975 TI - Oxidative damage to lens membranes induced by metal-catalyzed systems. AB - The oxidative effect of hydrogen peroxide, ascorbic acid and glucose in the presence of transition metals was studied. Incubation of bovine lens membranes with hydrogen peroxide/metal, ascorbic acid/metal and glucose/metal systems resulted in a significant augmentation of thiobarbituric acid-reactive substance content in the membranes. Presence of alpha-tocopherol decreased the extent of oxidative damage. This oxidation was found to be mediated by hydroxyl radicals. Dependence of hydroxyl radical generation upon the buffer used has been observed. In all experiments Cu(II) ions exhibited a higher efficiency compared to Fe(II) ions. PMID- 8727976 TI - Effect of galactose on alpha-crystallin. AB - Chromatographic separation of alpha-crystallin incubated with [3H]-labelled galactose showed the radioactivity to be concentrated in the low molecular mass subunits (20 and 40 kDa). The effect of glycation on the structural organization of alpha-crystallin was evaluated by FPLC analysis of native (pH 6.8 and 8.2) and glycated protein in dissociating conditions. Results suggest that the glycation acts on the protein surface by altering its charge distribution. PMID- 8727977 TI - Fatty acid composition of membrane phospholipids of cataractous human lenses. AB - The effect of lipid peroxidation on fatty acid composition in membrane phospholipids of transparent and cataractous lens nuclei was investigated. The preliminary results demonstrate a trend toward an increase in the content of palmitic acid with a decrease in the content of oleic acid, linolenic acid and docosahexaenoic acid in the opaque nuclei as compared to the transparent ones of senile cataractous lenses. PMID- 8727978 TI - Olfaction: what's new in the nose. PMID- 8727979 TI - Perireceptor events in olfaction. AB - Before reaching olfactory receptor neurons, odorant molecules have to cross an aqueous interface: the nasal mucus in vertebrates and the sensillar lymph in insects. Biochemical interactions taking place between odorants and the elements of these phases are called perireceptor events. Main protein constituents of these media, in both insects and vertebrates, are OBPs (odorant-binding proteins). Another class of proteins active in the olfactory perireceptor area includes odorant-degrading enzymes. The structure and the properties of these major proteins, with particular reference to OBPs, are reviewed and their role in olfactory transduction is discussed. PMID- 8727980 TI - Information processing in mammalian olfactory system. AB - In recent years, considerable progress has been made in understanding how the olfactory system uses neural space to encode sensory information. In this review, we focus on recent studies aimed at understanding the organizational strategies used by the mammalian olfactory system to encode information. The odorant receptor gene family is discussed in the context of its genomic organization as well as the specificity of olfactory sensory neurons. These data have important consequences for the mechanisms of odorant receptor gene choice by a given sensory neuron. Division of the olfactory epithelium into zones that express different sets of odorant receptors is the first level of input organization. The topographical relationship between periphery and olfactory bulb represents a further level of processing of information and results in the formation of a highly organized spatial map of information in the olfactory bulb. There, local circuitry refines the sensory input through various lateral interactions. Finally, the factors that may drive the development of such a spatial map are discussed. The onset of expression and the establishment of the zonal organization of odorant receptor genes in the epithelium are not dependent upon the presence of the olfactory bulb, suggesting that the functional identity of olfactory sensory neurons is determined independently of target selection. PMID- 8727981 TI - Second messenger signaling in olfactory transduction. AB - Olfactory receptor neurons respond to odorants with G-protein mediated increases in the concentration of cyclic adenosine 3',5'-monophosphate (cAMP) and/or inositol 1,4,5-trisphospahte (InsP3). These two second messengers directly regulate opening of cAMP- and InsP3-regulated conductances localized to the apical transduction compartments of the cell (cilia and olfactory knob). In the presence of physiological concentrations of extracellular Ca2+, these second messenger regulated conductances mediate influx of Ca2+ into the olfactory neuron resulting in large, localized increases in intracellular Ca2+ ([Ca2+]i). A significant advance in our understanding of the molecular mechanisms of olfaction is the recent realization that this increase in [Ca2+]i plays an important role as a "third messenger" in olfactory transduction. Second messenger dependent increases in [Ca2+]i cause opening of ciliary Ca(2+)-activated Cl-, cation and/ or K+ channels that can carry a large percentage of the generator current, thus amplifying the signal substantially. As a result of this sequence of events, the generator potential in olfactory neurons can be depolarizing, leading to excitation of the neuron, or hyperpolarizing, leading to suppression of basal action potential firing rate. This dual effect of odorants on olfactory neurons may play an important role in quality coding and in the ability to detect low concentrations of odorants, particularly in complex mixtures. PMID- 8727982 TI - Gaseous second messengers in vertebrate olfaction. AB - Gaseous monoxides such as nitric oxide (NO) and carbon monoxide (CO) are now recognized as important messengers in the nervous system. The enzymes generating these compounds are highly expressed in the olfactory system, including the epithelium and the main and accessory bulbs. Although the physiological roles of these molecules is still not entirely clear, some important new data has recently emerged. Alternate pathways for NO action, possible interactions between NO, CO, and intermediate proteins, and evidence suggestive of important roles for these molecules in development and regeneration are reviewed here. Of particular interest is the possible modulatory role of NO or CO in the transduction process, an area in which there has been an explosive growth in new data. Although it is clear that NO and CO are integral to the functioning of the olfactory system, it is equally obvious that many of the potential roles have yet to be clearly defined. PMID- 8727983 TI - Functional role of receptor neurons in encoding olfactory information. AB - In the present review we have considered the properties of the olfactory receptor neurons and discuss the strategy these cells use to perform their signaling task. Special emphasis is laid on the mechanisms for setting the membrane potential at rest and the mechanisms that the cell can use to respond with action potentials to significant stimuli only. We demonstrate that the firing properties of the receptor neurons depend upon the initial level of the membrane potential. We present the idea that the olfactory glomerulus can function as a unit in olfactory processing. In this perspective the olfactory receptor neuron is a subunit of the olfactory glomerulus. PMID- 8727984 TI - Neurogenesis and cell death in olfactory epithelium. AB - The olfactory epithelium (OE) of the mammal is uniquely suited as a model system for studying how neurogenesis and cell death interact to regulate neuron number during development and regeneration. To identify factors regulating neurogenesis and neuronal death in the OE, and to determine the mechanisms by which these factors act, investigators studied OE using two major experimental paradigms: tissue culture of OE; and ablation of the olfactory bulb or severing the olfactory nerve in adult animals, procedures that induce cell death and a subsequent surge of neurogenesis in the OE in vivo. These studies characterized the cellular stages in the olfactory receptor neuron (ORN) lineage, leading to the realization that at least three distinct stages of proliferating neuronal precursor cells are employed in generating ORNs. The identification of a number of factors that act to regulate proliferation and survival of ORNs and their precursors suggests that these multiple developmental stages may serve as control points at which cell number is regulated by extrinsic factors. In vivo surgical studies, which have shown that all cell types in the neuronal lineage of the OE undergo apoptotic cell death, support this idea. These studies, and the possible coregulation of neuronal birth and apoptosis in the OE, are discussed. PMID- 8727985 TI - Olfactory information processing in the brain: encoding chemical and temporal features of odors. AB - A fundamental problem in studying the neural mechanisms of odor recognition and discrimination in the olfactory system lies in determining the features or "primitives" of an odor stimulus that are analyzed by glomerular circuits at the first level of processing in the brain. Several recent studies support the idea that it is not simply the molecular features of odors that contain important information, but also the intermittent pattern of their presentation to the olfactory epithelium that helps determine the behavioral response to odor. PMID- 8727986 TI - Multiple factors shape development of olfactory glomeruli: insights from an insect model system. AB - The antennal system of the moth Manduca sexta is a useful model for studies of the development of olfactory glomeruli, the complex synaptic structures that typically underlie the initial processing of olfactory input in vertebrates and invertebrates. In this review, we summarize cellular events in the construction of glomeruli in Manduca and highlight experiments that reveal factors that influence glomerulus development. By methodically manipulating each of various cell types, both neuronal and glial, that contribute to glomerular architecture, we have found that: olfactory receptor axons lay a template for developing glomeruli, stabilization of the template by glial cells is necessary to permit subsequent steps in development of the glomeruli, and the hormone that regulates adult development causes production of adequate numbers of glial cells. Neither electrical activity nor the presence of a serotonin-containing neuron that persists throughout development is required for a glomerular pattern to develop; these factors might, however, influence the synaptic organization of individual glomeruli. PMID- 8727988 TI - Functional organization of olfactory system. PMID- 8727987 TI - Oscillations and gaseous oxides in invertebrate olfaction. AB - Olfactory systems combine an extraordinary molecular sensitivity with robust synaptic plasticity. Central neuronal circuits that perform pattern recognition in olfaction typically discriminate between hundreds of molecular species and form associations between odor onsets and behavioral contingencies that can last a lifetime. Two design features in the olfactory system of the terrestrial mollusk Limax maximus may be common elements of olfactory systems that display the twin features of broad molecular sensitivity and rapid odor learning: spatially coherent oscillations in the second-order circuitry that receives sensory input; and involvement of the interneuronal messengers nitric oxide (NO) and carbon monoxide (CO) in sensory responses and circuit dynamics of the oscillating olfactory network. The principal odor processing center in Limax, the procerebrum (PC) of the cerebral ganglion, contains on the order of 10(5) local interneurons and receives both direct and processed input from olfactory receptors. Field potential recordings in the PC show an oscillation at approximately 0.7 Hz that is altered by odor input. Optical recordings of voltage changes in local regions of the PC show waves of depolarization that originate at the distal pole and propagate to the base of the PC. Weak odor stimulation transiently switches PC activity from a propagating mode to a spatially uniform mode. The field potential oscillation in the PC lobe depends on intercellular communication via NO, based on opposing effects of reagents that decrease or increase NO levels in the PC. Inhibition of NO synthase slows the field potential oscillation, while application of exogenous NO increases the oscillation frequency. A role for CO in PC dynamics is suggested by experiments in which CO liberation increases the PC oscillation frequency. These design features of the Limax PC lobe odor processing circuitry may relate to synaptic plasticity that subserves both connection of new receptors throughout the life of the slug and its highly developed odor learning ability. PMID- 8727989 TI - Effect of ractopamine on growth performance, carcass composition, and cutting yields of pigs slaughtered at 107 and 125 kilograms. AB - At approximately 68 kg live weight, crossbred barrows and gilts (n = 144) were allocated to be fed to one of two weight end points (107 kg and 125 kg). Pigs from each weight group were treated with Ractopamine (RAC) (0, 10, or 20 ppm; n = 24/ treatment for the last 40 kg of gain. Feed consumption and weight gain were measured. Pigs were slaughtered and carcass measurements made at 24 h postmortem. Carcasses were fabricated into wholesale, trimmed wholesale, and boneless wholesale cuts for cutting yields. Hams were separated into muscle, fat, and bone. The RAC improved growth characteristics and carcass characteristics. Pigs fed RAC had increased (P < .01) average daily gain and improved (P < .01) feed:gain ratio over controls in each weight group. Carcasses from pigs treated with RAC had larger (P < .01) longissimus muscle area and reduced (P < .01) fat at the 10th rib. Cuts from 125-kg pigs were generally heavier than those from 107 kg pigs. The RAC increased (P < .05) the boneless cut weights of both weight groups. Percentage of dissected lean from the hams of RAC-treated pigs was (P < .05) higher than that of controls. Few consistent differences were observed between the 10 and 20 ppm of RAC treatments. Results from this study indicate that RAC had positive effects on the growth characteristics, carcass characteristics, and carcass cutting yields of pigs representative of the broad spectrum of market weights. PMID- 8727990 TI - Variability of energy metabolism and nuclear T3-receptors within the skeletal muscle tissue of pigs different with respect to the halothane gene. AB - Energy metabolism of skeletal muscle tissue of pigs growing from approximately 12 to 18 kg (12 homozygous halothane negative, HH; 16 heterozygotes, Hh; 17 homozygous halothane susceptible, hh) was measured in vivo using 31P nuclear magnetic resonance (NMR) spectroscopy. Data for intracellular pH, phosphocreatine (PCr), phosphomonoesters (PME), and ATP were analyzed by canonical discriminant analysis, an artificial neural network approach, and analysis of variance. Within the hh pigs, two subpopulations could be distinguished before the application of halothane treatment. Some of the hh pigs had a high PME concentration in the biceps femoris muscle (hh(pme+)), whereas others had a low concentration (hh(pme )) (2.18 +/- .12 for hh(pme+) vs 1.68 +/- .12 mM for hh(pme-), P < .004). The hh(pme+) pigs were statistically different from HH pigs for pH (P < .03), PME (P < .004), and PCr (P < .008) before halothane treatment. The hh(pme-) pigs were not different from the Hh and HH pigs with respect to PME when measured before halothane treatment (P > .05). However, intracellular pH (P < .03) and PCr (P < .008) of the hh(pme-) pigs were different from those of HH pigs (7.15 vs 7.19 for pH and 38.7 vs 35.1 for PCr, respectively). When combining intracellular pH, PME, and PCr within a canonical discriminant analysis, all were measured before halothane treatment, Hh pigs were found to be different from HH pigs (Mahalanobis distance different from zero, P < .02). In a second experiment, growth rate, depth of longissimus muscle, and maximal binding capacity of nuclear T3-receptors of skeletal muscle tissue were different (P < .05, P < .002, and P < .02, respectively) among pigs selected from the same genetic lines. Of the variability in depth of the longissimus muscle, 22% was explained by variability in maximal binding capacity of nuclear T3-receptors. These results, if confirmed with a large number of pigs, might open new possibilities for selection procedures for leanness because, with respect to halothane susceptibility, a shift between genotypic and phenotypic variability was observed. PMID- 8727991 TI - Breeding season and aspects of reproduction of female goats. AB - Reproductive data were collected on 608 female goats and their 1,147 offspring, involving 20 herds, from different geographical locations in Georgia for 3 yr. Results for seven breeds and a dairy crossbred revealed that most goats bred seasonally, commencing approximately in late June and reaching a peak in September to November. However, the Pygmy had an unusual peak of mating activity during summer (July). Gestation period ( +/- SE) was 150.6 +/- 2.64 d. Pygmies had the shortest gestation period, whereas Toggenburgs had the longest. Gestation period decreased as the litter of size of the doe increased (b = -.92 d/kid, P < .001) and increased slightly with increasing parity (b = .22 d/parity). December and January matings had the shortest gestation period. Litter size was 1.85 +/- .67, with twins being the most prevalent litter size. Litter size varied among breeds. The litter size increased with mating weight of the doe for most breeds (litter size increased approximately .02 kids/kg of mating weight). Birth weight was 3.24 +/- .64 kg and varied among breeds; Pygmy kids were lightest (1.7 kg) and Toggenburgs were heaviest (3.9 kg). Males were heavier than female kids. Birth weight decreased with the size of litter (approximately .45 kg/kid, P < .001). PMID- 8727992 TI - Detection of estrus by visual observation and radiotelemetry in peripubertal, estrus-synchronized beef heifers. AB - The effectiveness of two estrus-detection methods (visual observation and radiotelemetric, pressure-sensitive, rump-mounted devices) was compared in peripubertal, crossbred yearling beef heifers. Heifers (n = 50) were fitted with a pressure-sensitive device affixed to their rumps to which a battery-operated radio transmitter was connected. Activation of the sensor sent a radiotelemetric signal to a microcomputer via a fixed radio antenna. Heifer identification, date, time, and duration of standing events were recorded. Estrus was synchronized by feeding melengestrol acetate (MGA) for 14 d and then injecting PGF2 alpha 17 d after the last daily dose of MGA. Following the injection of PGF2 alpha, heifers were observed visually for signs of estrus at 0730 and 1630 (45 min each). Estrus was detected in 41 of 49 heifers (one lost the device) by visual observation and(or) the radiotelemetric device. Number of standing events during estrus, determined by the radiotelemetric device, averaged 50.1 +/- 6.4 per heifer, with the duration of estrus ranging from 2.6 to 26.2 h (average = 14 +/- .8 h). Visual observation failed to detect 11 of 41 heifers (37%) that were detected by the radiotelemetric device. Heifers with fewer standing events (19.3 vs 60.5; P < .001) and estrus of shorter duration (8.4 vs 15.6 h; P < .001) were those not identified by visual observation. Based on intervals between the first detected standing event of estrus and time of insemination, a wide window of fertility seems to exist in some heifers. Radiotelemetric devices were useful in identifying a greater (P < .05) proportion (100%) of heifers in estrus (increased efficiency) compared with visual observation (73%). However, accuracy of both methods was similar. PMID- 8727993 TI - Effects of ionophore feeding and anthelmintic administration on age and weight at puberty in spring-born beef heifers. AB - Two hundred eighty-eight heifer calves in two 1-yr replications were used to test the hypothesis that ionophore (ION) feeding, anthelmintic (ANT) administration, or the combination (COMB) would cause heifers to express pubertal estrus at a younger age than control (CONT) heifers. Heifers were assigned randomly within three weight blocks in a randomized complete block design with a 2 x 2 factorial arrangement of treatments. A high-fiber diet was fed for 172 d during yr 1 (Y1) and 199 d during yr 2 (Y2). Monensin sodium (200mg.animal-1.d-1) was fed to ION and COMB treatments and ivermectin (200 micrograms/kg of BW) was given to ANT and COMB groups at weaning and 84 d later. Rate of gain was monitored every 21 d and diet intake was adjusted to obtain equal rates of gain across treatments. Fecal samples were collected from one-third of the heifers in each group and analyzed for internal parasite egg counts every 21 d. Serum progesterone samples taken twice weekly were used to estimate onset of puberty. There was no treatment x year interaction in any parameter measured and the data were pooled across both years. Total weight gain was similar (P > .10) for all treatments. Fecal egg output decreased (P < .01) after initial application of anthelmintic. The ANT, ION, and COMB heifers had greater (P < .05) gain:feed ratios than did CONT (.132, .142, and .148 vs .125 kg for ANT, ION, COMB, and CONT, respectively). Age and weight at puberty were different (P < .05) for ION (425 d, 351.8 kg), COMB (425 d, 349.5 kg), and ANT (424 d, 348.8 kg) compared with CONT (433 d, 362.5 kg). There was no difference (P > .10) in age and weight at puberty between COMB, ION, or ANT heifers. During Y1, but not Y2, there were a greater (P < .05) number of COMB, ION, and ANT heifers pubertal before the breeding season. Additionally, there was a tendency (P = .12) for higher first-service conception in the COMB, ION, and ANT heifers than in the CONT heifers. Final pregnancy rates among treatments were similar (P > .10). PMID- 8727994 TI - Effect of space allowance on barrow performance to 136 kilograms body weight. NCR 89 Committee on Management of Swine. AB - Two experiments were conducted to examine the space requirements of barrows grown to 136 kg BW. In Exp. 1, barrows (initial BW 19.5 kg) were allowed .65, .84, or 1.02 m2/pig floor space in a fully slatted facility with 12 pigs/pen. In Exp. 2, conducted at three experiment stations in the north central region of the United States (Iowa, Indiana, Nebraska), barrows (initial BW 55.5 kg) were allowed .65, .93, or 1.20 m2/pig floor space in either partially or totally slatted facilities. In Exp. 1, there was a linear (P < .05) decrease in ADG for the first 29-d period with increasing space. Overall, ADG responded in a quadratic (P = .09) manner to increasing space allocation (.841, .855, and .824 kg/d for the .65, .84, and 1.02 m2 treatments, respectively) with a linear (P = .06) decrease in gain:feed ratio (.322, .318, and .312). Space allocation had no effect (P > .10) on ADFI, last rib fat depth, or rate of lean gain. In Exp. 2, ADG improved linearly (P = .08) with increasing space (.836, .857 and .865 kg/d for the .65, .93, and 1.20 m2 treatments, respectively), with a quadratic (P = .07) response in gain:feed ratio (.270, .265, and .268, respectively). There was no effect of space allocation on uniformity of gain within a pen as measured by coefficient of variation for pig weights within a pen at the time of first pig removal. The results of these experiments suggest that performance of barrows grown to 136 kg BW was maximized with .84 to 1.0 m2 of floor space per pig. PMID- 8727995 TI - Effect of selection for size of testes in boars on semen and testis traits. AB - The objective of this study was to evaluate the effects of selection for increased size of testes on semen and testis characteristics. Boars from a line selected for increased size of testes at 150 d of age (TS, n = 25) and a randomly selected control line (C, n = 18) were used. Semen was collected three times per week for 3 wk (TRT1) then daily for 3 wk (TRT2) beginning when boar age averaged 276 d. It was followed by a 4-wk period of one collection per week and then the TRT1-TRT2 sequence was repeated. After 64 collections, boars were castrated and testes were evaluated for sperm numbers. Volume of semen, percentage of motile sperm cells, and percentage of abnormal cells in the semen did not differ (P > .05) between lines. Concentration of sperm cells in the semen was greater for TS boars at TRT 1 (35.0 million cells/mL, P < .10) and TRT 2 (29.5 million cells/mL, P < .01). Number of sperm cells per ejaculate was greater for TS boars (6.1 and 4.3 billion cells for TRT 1 and TRT 2, respectively; P < .05). Percentage of abnormal cells was less (P < .05) for boars at the younger ages. Concentration of sperm cells was 16 to 18 million cells/mL semen greater (P < .05) at the older ages. Significant differences in the shapes of the response curves with increasing collection number across lines and ages were found. These differences were small and not considered to be important. Line differences in testicular and epididymal sperm numbers and in daily sperm production were consistent with differences in semen traits. Repeatabilities of semen traits were between .16 and .74. Selection for increased size of testes can be used to improve the reproductive capacity of boars used for artificial insemination. PMID- 8727996 TI - Genetic parameter estimates of live animal ultrasonic measures of retail yield indicators in yearling breeding cattle. AB - Ultrasonic measures of backfat thickness (BFT) and longissimus muscle area (LMA) taken on Angus bulls (n = 805) and heifers (n = 877) ranging in age from approximately 8 to 20 mo in a production setting in western Nebraska were used to estimate genetic (co)variances. Further information used in the analyses, including weaning weight (WWT), postweaning ADG, and scrotal circumference (SC), was obtained from the American Angus Association, St. Joseph, MO, for these individuals and for animals from the same herd and contemporary groups. Data were analyzed using single-trait, sire model, restricted maximum likelihood (REML) procedures to estimate starting variances for later two-trait analyses. These two trait analyses were done to estimate variance components for WWT coupled with BFT, LMA, SC, and ADG, also using REML procedures, but with a sire-maternal grandsire model for WWT and a sire model for the other trait(s). Heritabilities for BFT, LMA, WWT direct, WWT maternal, SC, and ADG were .56, .11, .19, .24, .56, and .51, respectively. Genetic correlations between WWT direct and WWT maternal, WWT direct and LMA, WWT maternal and LMA, WWT maternal and BFT were -.57, .42, .01, and -.69, respectively. Genetic parameters from this study were used to calculate possible genetic change with a typical selection scenario, and it was shown that among WWT, BFT, and LMA, BFT could be affected the most by selection, relative to its mean. PMID- 8727997 TI - Behavioral and pituitary adrenal-axis responses of dairy cows to social isolation and deprivation of lying down. AB - The behavior and plasma concentrations of ACTH and cortisol were studied in 30 Friesian cows kept in tie stalls and assigned to three treatments: control (C), deprivation of lying down from 0900 to 1600 and 2200 to 0500 (D), and social isolation (I). Behavior of the cows was observed before and after 4 and 8 wk of treatment. The D- and I-cows showed increased frequency of transitions between different behavioral activities (P < .05). In D- and I-cows the frequency of grooming (P < .01) and idling (P < .001), and the amount of leaning (P < .001) increased. In D-cows the frequency of eating was also increased. The behavior and cortisol response to two novel arena tests performed on two consecutive days after 22 d of treatment suggests that social isolation and deprivation of lying change cows' reactions to a novel environment. After 23 d of treatment, concentrations of ACTH and cortisol were measured during 7.5 h. On d 24, cortisol concentrations were determined at time -.5, 0, .5, 1, 2, 3, 4, and 5 h after i.v. administration of ACTH. Cortisol concentrations did not differ among treatment groups in the baseline series and after ACTH administration. In D-cows, ACTH concentration was increased in part of the baseline series (P < .05). The results suggest that the socially isolated cows were frustrated or tried to compensate for a lack of stimulation and that repeated deprivation of lying down is aversive to cows. PMID- 8727998 TI - Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and western blotting comparisons of purified myofibrils and whole muscle preparations for evaluating titin and nebulin in postmortem bovine muscle. AB - Purified myofibril (MF) and homogenized whole muscle (WM) samples were prepared from A maturity market steers. Samples were removed at 0, 1, 3, 7, 14, and 28 d postmortem. The MF and WM samples from all steers were analyzed by SDS-PAGE (5% gels) and by Western blot analysis using monoclonal antibodies to titin and nebulin. The rates of degradation of the intact forms of titin and nebulin, with regard to differences dependent on sample type (MF vs WM), were examined. The results showed that there was very little difference in the rate of postmortem degradation of the intact form of titin or of intact nebulin with respect to the two types of samples examined. Analysis of MF and WM preparations revealed that titin and nebulin were progressively degraded, each at its own rate, with nebulin degrading faster, as postmortem storage time increased. Examination of MF and WM samples showed that the intact form of titin (T1) was absent at the same time postmortem in both sample types. Intact nebulin was not detected in MF and WM preparations at the same time postmortem with respect to sample type examined. Our results indicate that either purified MF or WM samples can be used satisfactorily to analyze the rate of degradation of the intact forms of both titin and nebulin. PMID- 8727999 TI - Yields, chemical composition, and value of beef shank tissues obtained using Baader processing. AB - This experiment was designed to determine yield of meat and sinew from beef shanks processed with a Baader desinewing machine and to determine whether this process added value to a beef carcass. Baader desinewing machines use belt pressure against a rotating, perforated steel drum to separate tissues. Boneless beef shanks had 9.8% fat and 14.1 mg/g of collagen. Using the Baader with a 5-mm drum, the first pass lean yield was 73.3% and had fat reduced to 7.1% and collagen to 10.5 mg/g. Second-pass lean yield through the 5-mm drum was 19.6% and had 16.1% fat and 13.8 mg/g of collagen, leaving 6.7% separated sinew. Using a 3 mm drum reduced first-pass lean yield to 66.1% and reduced fat content to 5.8%. Second-pass lean yield, using 3- and 5-mm drums, was 26.1% and had 18.6% fat and 27.8 mg/g of collagen with 6.8% sinew. Desinewed lean is worth more than whole shanks. Furthermore, 95% lean is worth more than 90% lean, and the sinew also has a salvage value. Upgrading shanks with this desinewing device can increase the value of a beef carcass by $2.01 using a 5-mm drum or by $3.20 using both 3- and 5-mm drums. PMID- 8728000 TI - Modification of early-postmortem muscle pH and use of postmortem aging to improve beef tenderness. AB - This study was conducted to determine whether modification of early-postmortem muscle pH and(or) control of postmortem aging time could be used to improve tenderness of strip loin (longissimus, LM), top sirloin (GM), and top round (SM) steaks. Muscle pH, at 1.5 and 3 h postmortem, was modified using three 60-Hz electrical stimulation (ES) treatments (240 V, 35 V, or combined use of 240 and 35 V). Vacuum-packaged sections of the LM, GM, and SM were aged for 6, 12, 18, or 24 d postmortem. Use of ES increased rate of postmortem muscle pH decline and decreased shear force of LM steaks but had no effect on tenderness of GM or SM steaks. Reducing 3-h pH of the LM reduced variability in LM shear force but had little effect on tenderness of GM or SM steaks. However, lower values for 24-h pH of the LM were associated with increased tenderness of all three cuts. Tenderness of all cuts also improved as length of the postmortem aging period was increased. For LM and SM steaks, the greatest improvements in tenderness were achieved by 12 d; however, tenderness of GM steaks continued to increase up to 24 d postmortem. PMID- 8728001 TI - Nutrient digestibilities of a corn-soybean meal diet as influenced by Bacillus products fed to finishing swine. AB - Twelve crossbred barrows (initial BW of 59.7 kg) were used in nutrient balance trials to investigate the influence of adding two Bacillus products (Biomate 2B and Pelletmate Livestock, Chr. Hansen's Biosytems) to a 14% CP corn-soybean meal diet (.64% lysine, .6% Ca and .5% P). A 3 x 3 Latin-square design with an extra period was used. Treatments were as follows: 1) basal diet; 2) basal + Biomate 2B (BAC1); and 3) basal + Pelletmate Livestock (BAC2). After a 7-d adaptation to metabolism crates, each 10-d period consisted of 5 d of diet adjustment followed by 4 d of total collection and 1 d for change-over. Both BAC1 and BAC2 were added at a level of .05% to supply approximately 3 million colony forming units per gram of diet. Pigs were fed twice daily at a daily rate of 3.2 to 3.5% of BW. Feeding BAC1 or BAC2 elevated (P < .001) fecal Bacillus spore counts at the end of each collection period, but the effect on number of coliforms and lactic acid bacteria was not as consistent; coliform counts tended to be decreased (P < .10) when BAC2 was fed, but not (P > .10) when BAC1 was fed. The number of lactic acid bacteria was increased (P < .05) when BAC1 was fed, but not (P > .10) when BAC2 was fed. Body weights were similar among all treatments for all periods. There was no carry-over or pretreatment effect (P = .50). Digestibility coefficients of DM, neutral detergent fiber, acid detergent fiber, ash, and N and retention of N were not affected (P = .40) by feeding Bacillus products. PMID- 8728002 TI - Performance and body composition of finishing gilts (45 to 85 kilograms) as affected by energy intake and nutrition in earlier life: I. Growth of the body and body components. AB - Forty-eight commercial hybrid gilts were used to determine the effects of energy intake from 20 to 45 kg on the relationship between energy intake and weight gain of the body and body components from 45 to 85 kg. Two groups of 24 gilts received a single diet either at 2.2 (restricted) or 3.7 (control) times maintenance (M) from 20 to 45 kg. From 45 to 85 kg the pigs were fed the same diet at one of six intake levels (1.7, 2.2, 2.7, 3.2, or 3.7 x M, or ad libitum). At 85 kg, the gilts were dissected into organs, lean tissue (trimmed major joints), and other carcass parts (fat fraction). In the restricted gilts, body and lean tissue gain between 45 and 85 kg increased curvilinearly with increasing energy intake from 432 to 1,412 g/d and from 228 to 507 g/d, respectively. In the control gilts, body and lean tissue gain increased from 394 to 1,201 g/d and from 238 to 508 g/d, respectively. The percentage of lean tissue in the carcass decreased curvilinearly with increasing energy intake from 62.9 to 56.5%, and from 62.5 to 53.9% in the restricted and control gilts, respectively. The restricted gilts gained on average 140 g/d faster (P < .001) and their carcass lean tissue content at slaughter was approximately 3% higher (P < .001). These differences increased with increasing energy level between 45 and 85 kg. The increased gain was largely the result of an increase in organ growth and gut contents and was only evident up to 65 kg. The higher lean content at 85 kg in previously restricted gilts was not the result of compensation in lean gain but was already present at the end of the restriction phase at 45 kg. PMID- 8728003 TI - Performance and body composition of finishing gilts (45 to 85 kilograms) as affected by energy intake and nutrition in earlier life: II. Protein and lipid accretion in body components. AB - Forty-eight commercial hybrid gilts were used to investigate the response relationships between energy intake and deposition of protein and lipid in body components of finishing pigs (45 to 85 kg) as affected by previous nutrition. Two groups of 24 gilts received a single diet either at 2.2 (restricted pigs) or 3.7 (control pigs) times maintenance (M) from 20 to 45 kg. From 45 to 85 kg, pigs from each of these two groups were fed the same diet at one of six intake levels (1.7, 2.2, 2.7, 3.2, or 3.7 x M, or adlibitum). At 85 kg, the gilts were dissected and analyzed. Protein content in the whole body decreased curvilinearly from 190 to 166 g/kg (P < .001) and lipid content increased from 116 to 210 g/kg (P < .001) with increasing energy intake. The mean content of protein was 6 g/kg higher and of lipid 29 g/kg lower (both P < .001) in previously restricted gilts. Protein (PD) and lipid deposition (LD) in the whole body, organs, lean, and fat tissue increased linearly (P < .001) with increasing energy intake. In the whole body, PD increased from 83 to 187 g/d and LD from 46 to 392 g/d. The respective response relationships in grams/day were PD = 21.8 + 3.78 x DEI and LD = -193 + 13.7 x DEI(DEI is digestible energy intake, MJ/d). The LD:PD ratio increased curvilinearly from .6 to 2.2. Previous nutrition had only small effects on PD and LD. Only PD in the organs was on average 7 g/d higher (P < .001) in previously restricted gilts. Thus, the higher protein content and the lower lipid content in previously restricted gilts was not primarily the results of compensation but was almost entirely present at the end of the restriction phase at 45 kg. PMID- 8728004 TI - Effects of previous grazing treatment and consumption of locoweed on liver mineral concentrations in beef steers. AB - Twelve Hereford steers (average BW = 231 kg) that had previously grazed native rangeland (Range) or irrigated winter wheat pasture (Wheat) were allowed to graze locoweed-infested rangeland from April 1 to June 9, 1994 (six steers/previous grazing treatment). Relative consumption level of locoweed and other forage classes was measured as observed bites per steer. Liver biopsy and whole blood samples were obtained from each steer before and after grazing. Liver samples were analyzed for several minerals by inductively coupled plasma-atomic emission spectroscopy, and whole blood samples were analyzed for Se. Liver concentrations of Ba (P < .001), Cd (P < .001), Ca (P < .01), Cr (P < .01), Ni (P < .001), Na (P < .01), and V (P < .001) were greater and concentrations of Mn (P < .09), P (P < .01), and K (P < .07) were less in Wheat than in Range steers. Liver concentrations of Fe, Mg, S, and Zn and whole blood Se concentrations did not differ (P > .10) between the two groups. Liver concentrations of Cr (P < .04) and Mn (P < .001) were less, and Fe concentrations were greater (P < .01), in samples taken after grazing than in samples taken before grazing of locoweed-infested range. Whole blood Se concentrations decreased (P < .01) from the beginning to the end of the grazing period, but this effect was not related (P > .15) to locoweed consumption. Changes in liver concentrations of minerals were compared relative to consumption levels of all forage classes in the locoweed-infested range. Liver concentrations of Cu decreased (r2 = .45; P < .02) as the percentage of bites consumed as locoweed increased, but concentrations after grazing locoweed-infested range were still within normal ranges. Changes in liver concentrations of other minerals were not related (P > .15) to consumption of locoweed. These data indicate that previous grazing history can have significant effects on liver mineral stores and that, under our conditions, consumption of locoweed by grazing beef steers altered liver Cu concentrations. Toxic effects of locoweed consumption would likely occur before Cu deficiency would be induced by grazing locoweed-infested range; hence, supplementation of Cu would seem unlikely to alter the course of locoweed toxicosis. PMID- 8728005 TI - Calcium decreases and parathyroid hormone increases in serum of periparturient mares. AB - Changes in serum concentrations of Ca and parathyroid hormone (PTH) may develop in periparturient mares, may be influenced by dietary Ca, and may be associated with changes in Ca concentration of mammary secretion. Milk and blood samples were taken from eight mares on Farm A and eight on Farm B for 10 d before parturition and from four mares on each farm for 5 d postpartum. Milk Ca was measured by two commercial tests. Serum samples were analyzed for PTH and total Ca in 16 mares and for ionized Ca in six (Farm A). Parturition was induced in eight mares on Farm A and four on Farm B; no significant difference in serum Ca or PTH was found between mares with induced and spontaneous foaling. Mean serum total Ca decreased from 12.5 mg/dL to a nadir of 11 mg/dL on d 2 postpartum, and mean PTH increased from 46 pg/mL to a peak of 186 pg/mL on d 2 postpartum. Serum total Ca concentrations were lower and serum PTH concentrations were higher in Farm A mares than in Farm B mares, probably reflecting dietary Ca (.35% of DM on Farm A and .55% on Farm B). The serum PTH peak and Ca nadir occurred on d 2 postpartum, 1 d later than reported previously in dairy cows. Milk Ca concentration increased progressively from 7 d before parturition; this increase preceded, so was not dependent on, prepartal changes in serum Ca and PTH. PMID- 8728006 TI - Endocrine and metabolic responses to realimentation in feed-restricted prepubertal gilts: associations among gonadotropins, metabolic hormones, glucose, and uteroovarian development. AB - Temporal relationships among plasma LH, FSH, metabolic hormones, and glucose concentrations were studied in nutritionally manipulated prepubertal gilts. Twenty-four growth-matched littermate gilts (73.6 +/- 1.2 kg) were fed at maintenance for 7 d (d 1 to 7). From d 8 to 14, littermates were divided between two treatments and were either fed twice daily to appetite or continued on feed restriction. Feeding to appetite increased episodic LH secretion during the first 6 h of sampling on d 8 (P < .05). Plasma FSH concentrations were not affected by treatment between d 8 and 14. Nocturnal increases in plasma LH (P < .01) and FSH (P < .002) concentrations were detected in feed-restricted gilts but not realimented gilts. Mean total plasma IGF-I concentrations gradually increased in gilts fed to appetite (d 12, P < .02; d 13 to 14, P < .001). Mean postprandial insulin concentrations were also greater in gilts fed to appetite (P < .01). Treatment differences in mean postprandial glucose concentrations on d 9 and 10 indicate that normoglycemia was established by d 9 in gilts fed to appetite. At slaughter on d 15, numbers of ovarian follicles (P < .01), follicular volume (P < .001), and uterine weight (P < .05) were greater in gilts fed to appetite. In conclusion, uteroovarian responses to realimentation after short-term feed restriction in prepubertal gilts are mediated by enhanced LH secretion. This rapid enhancement of LH secretion in response to dietary repletion might have been mediated by changes in glucose and insulin status. Ovarian responses to gonadotropins may also be potentiated by observed increases in plasma glucose, insulin, and IGF-I. During feed restriction, a distinct diurnal rhythm in gonadotropin secretion was evident. PMID- 8728007 TI - Identification of matrix metalloproteinases and metalloproteinase inhibitors in bovine corpora lutea and their variation during the estrous cycle. AB - Corpora lutea (CL) were collected from cattle to study key physiologic events in angiogenesis. Our objective was to evaluate the activity of matrix metalloproteinases (MMP) and endogenous inhibitors. Corpora lutea were collected 2, 4, 6, 8, 10, 12, 14, and 16 d (n = 3/d) after estrus was first detected. In zymograms, a band of protein migrating at a relative molecular mass (M(r)) of 98 kDa was increased early in the cycle; a M(r) = 88 kDa band was detectable on all days. The molecular weights of these proteins are consistent with the MMP-9 family members. In all samples, a band of enzyme activity was detected at M(r) = 62 kDa, and another band of lesser density was detected at M(r) = 60 kDa. The molecular weights of these proteins are consistent with the MMP-2 family members. An immunoreactive band, detected in all samples with equal density, migrated between M(r) = 27 and 29 kDa, as did the tissue inhibitor of metalloproteinase (TIMP-1) standard. A second band, which was less dense in samples from d 2 through 6, migrated at M(r) = 19 kDa, as did the TIMP-2 standard. A third band was detected in all samples; it migrated at M(r) = 35 kDa, as did the cartilage derived inhibitor (CDI) standard, and was less dense in d 8 and d 12 through 16 samples. In summary, MMP (gelatinases) and MMP inhibitors are present in developing luteal tissue, and the M(r) = 98 kDa enzyme, CDI, and TIMP-2 varied during the estrous cycle. PMID- 8728008 TI - Plasma and milk urea nitrogen in relation to pregnancy rate in lactating dairy cattle. AB - The objectives of this study were to relate concentrations of plasma (PUN) and milk (MUN) urea nitrogen to pregnancy rate in dairy cows and compare various methods of analysis and preparation of milk for measuring MUN. In two experiments, blood or milk samples were collected on the day of AI from Holstein cows (n = 160 and n = 155, respectively). Three methods of MUN analysis were compared. Two laboratory chemical procedures yielded similar results, whereas a quick dipstick method overestimated chemical analyses. Before and after milking strip samples had MUN concentrations equivalent to those in composite milk. Concentrations of PUN or MUN greater than 19 mg/dL were associated with decreased (P < .02) pregnancy rates (18 and 21 percentage point reduction in the two experiments). In two subset groups of cows (n = 51 and n = 23, respectively), plasma progesterone or MUN concentrations were monitored during the 5-d period after AI. Plasma progesterone concentrations increased similarly during the period for cows divided into low vs high PUN but were greater in pregnant than in nonpregnant cows on d 4 and 5 (P < .04). The MUN concentrations showed low within cow variation (CV = 8%) but were lower in pregnant cows and had a decreasing trend over time compared with nonpregnant cows (P < .05). Based on this study, plasma and milk will yield similar results for monitoring urea nitrogen in dairy cows; PUN and MUN concentrations > 19 mg/dL were associated with approximately a 20 percentage point decrease in pregnancy rate after AI in lactating dairy cattle. PMID- 8728009 TI - Effects of growth hormone, prolactin, insulin-like growth factors, and gonadotropins on progesterone secretion by porcine luteal cells. AB - Growth hormone (GH) and IGF-I have receptors within the corpus luteum (CL) and stimulate CL function. Our objective was to investigate the effects of GH, prolactin (PRL), IGF-I, IGF-II, LH, and FSH on progesterone secretion by porcine luteal cells during mid-pregnancy. Gilts (crossbred Yorkshire/Landrace) were slaughtered on d 44 of pregnancy and CL were collected. Large and small luteal cells (LLC and SLC, respectively) were obtained from dissociated CL and separated by elutriation. Luteal cells were incubated with 0, 1, 10, or 100 ng/mL of GH, PRL, IGF-I, IGF-II, LH, and FSH or combinations of 10 ng/mL of these reagents for 24 or 48 h. Culture media were harvested and concentrations of progesterone analyzed by radioimmunoassay. Growth hormone, PRL, and IGF-I increased (P < .05; 100 ng/mL dose) concentrations of progesterone in media of LLC. Insulin-like growth factor-II, LH, and FSH had no effect on progesterone in LLC cultures. In SLC cultures, GH, PRL, IGF-I, IGF-II, and FSH failed to stimulate progesterone secretion, whereas LH increased progesterone secretion (linear effect of dose; P < .05). Combinations (10 ng/mL each hormone) of GH and IGF-I or PRL and IGF-I increased progesterone secretion by LLC compared with control, GH, PRL, or IGF-I alone (P < .05). Similar combinations of GH or PRL with IGF-I had no effect on SLC. Conclusions are that GH and PRL are stimulatory to progesterone secretion by LLC (location of GH receptor) and SLC are responsive to LH during mid-pregnancy. Both GH and PRL are synergistic with IGF-I for increased progesterone secretion. PMID- 8728010 TI - Responses of porcine corpora lutea to somatotropin administration during pregnancy. AB - The effects of somatotropin (ST) on functions of porcine corpora lutea (CL) during pregnancy were investigated. Twenty-four crossbred (Yorkshire/Landrace) gilts from d 30 to 43 of pregnancy were injected daily with 5 mg of recombinant porcine somatotropin (rpST; n = 12) or 1 mL of saline (control, n = 12). Blood was collected on d 30, 37, and 43 for analyses of plasma progesterone. Gilts were killed on d 44 of pregnancy, and mRNA were isolated from CL, ovary, and liver. Messenger RNA expression for LH receptor, FSH receptor, ST receptor, 3 beta hydroxysteroid dehydrogenase (3 beta-HSD), and cytochrome P450 side-chain cleavage enzyme (SCC) were measured. Liver, CL, and ovary contained a 4.7-kb mRNA of ST receptor, but the liver contained more mRNA for ST receptor than did CL or ovary (.97 +/- .18, .47 +/- .04, and .25 +/- .04 units, respectively). There were two variants of LH receptor mRNA in CL (6.8 and 4.4 kb). The CL also contained a 1.8-kb mRNA of SCC and a 1.7-kb mRNA of 3 beta-HSD. No FSH receptor mRNA was detected in CL of the pig. The rpST treatment did not affect the mRNA level of ST receptor, 3 beta-HSD, SCC, or 4.4-kb mRNA of the LH receptor. The 6.8-kb mRNA for the LH receptor was decreased (P < .05) by rpST (.56 +/- .04 vs .78 +/- .05 units). Furthermore, concentrations of plasma progesterone decreased (P < .001) in gilts treated with rpST. Decreased luteal function was associated with decreased expression of LH receptor in rpST-treated gilts. The luteotropic effects of ST observed in vitro do not necessarily occur in vivo when gilts are administered rpST during pregnancy. PMID- 8728011 TI - Glucose tolerance of pregnant sows is related to postnatal pig mortality. AB - An experiment was conducted on three farms to study the relationship between the results of an oral glucose tolerance test on d 104 +/- 4 of pregnancy and subsequent reproductive performance of 104 multiparous sows. After an overnight period without feed, sows were fed (3 g/kg BW.75) glucose, and blood samples were taken for analyses of glucose at -10, 10, 20, 30, 40, 50, 60 70, 80, 90, 105, and 120 min after administration of glucose. Glucose concentrations before glucose administration ranged from 2.0 to 3.6 mmol/L. Sows with lesser pretest glucose concentrations had longer (P < .01) durations of pregnancy (corrected for number of live pigs born) and heavier (P < .05) pigs at birth (corrected for farm and number of live pigs born). Peak glucose concentrations and areas under the curves (i.e., first 70 min and entire 120 min) ranged from .1 to 4.4 mmol/L, -40 to 211 (mmol/L).min, and -95 to 247 (mmol/L).min, respectively. Sows with greater (P < .01) peak concentration of glucose and greater area under the curve (first 70 min [P < .01] and entire 120 min [P < .01]) had increased pig mortality during the first 7 d after farrowing (corrected for live weight of the sows). In 83 sows, a sample of morning urine (before feeding) was collected and tested for glucose and ketone bodies. Neither glucose nor the ketone body acetoacetic acid was detected in the urine. The results indicate that sows that are less glucose-tolerant have greater pig mortality. PMID- 8728012 TI - The effects of bovine somatotropin (bST) and porcine somatotropin (pST) on growth factor and metabolic variables in horses. AB - Effects of exogenous pST and bST on metabolic and growth factor variables were examined in three studies with lighthorse mares (455 to 545 kg). In Study 1, eight mares received five s.c. injections of bST or pST (30 mg/d). In Studies 2 and 3, five mares received one s.c. injection of a prolonged release formulation designed to deliver 500 mg of bST (Study 2) or pST (Study 3) over 14 d. Blood samples were collected for several days before injection to establish baseline values, at frequent intervals during treatment, and for several days thereafter. In all studies, blood urea nitrogen concentrations were decreased (P < .001) and insulin-like growth factor I (IGF-I) concentrations were increased (P < .001) within 48 h after bST or pST injection relative to pretreatment values. Similarly, insulin and glucose were increased (P < .001) relative to pretreatment values, after bST or pST administration. In Studies 2 and 3, circulating ST concentrations were increased (P < .001) for at least 14 d after injection, despite severe local tissue reactions at the prolonged release formulation injection site. Insulin-like growth factor I ligand blotting of serum revealed bands with molecular weights (MW) of 45, 32, 30, and 18 kDa, and two bands of > 96 kDa. These results indicate that 1) bST and pST are biologically active in horses, which respond metabolically to exogenous ST in a manner similar to other mammalian species, 2) circulating IGF binding proteins are present in horses, and 3) the commercially available dairy cow product POSILAC (Monsanto, St. Louis, MO) is not appropriate for the delivery of bST in horses due to injection site reactions accompanying the administration of the oil-based prolonged release formulation. PMID- 8728013 TI - Circadian changes in net nutrient fluxes across the portal-drained viscera, the liver, and the hindquarters in preruminant calves. AB - The objective of this study was to describe in preruminant calves the circadian patterns in net nutrient fluxes across the portal-drained viscera, the liver, and the hindquarters and to relate them to previously published variations in tissue energy expenditure. In vivo arterio-venous techniques were used, and animals were fed a conventional milk replacer. In the portal-drained viscera, net glucose absorption occurred 1 to 5 h postprandially with some lactate release. Arterial plasma non-esterified fatty acid (NEFA) concentrations decreased subsequently, as well as net NEFA uptake by the portal-drained viscera. Triglyceride absorption that occurred 1 as well as 7 or 8 h postprandially did not take place via the portal vein. Changes in energy expenditure of the portal-drained viscera did not correlate with changes in net nutrient fluxes. In the liver, the maximum contribution of lactate to hepatic glucose production was 20%. The NEFA and triglycerides were taken up by the liver, whereas urea was released. Only increases in NEFA uptake could partly explain the postprandial rise in hepatic energy expenditure. In the hindquarters, up to 48% of the glucose taken up could be converted into lactate; the remaining balance was possibly oxidized, thereby contributing as much as 33% to energy expenditure. No hourly patterns were noted in net nutrient fluxes through hindquarters. In conclusion, in preruminants, contribution of nutrients to oxidation differed from that noted in ruminants and seemed to vary with nutrient absorption and availability patterns, especially in the portal-drained viscera. PMID- 8728014 TI - Effects of dietary concentrate level on nutrient absorption, liver metabolism, and urea kinetics of beef steers fed isonitrogenous and isoenergetic diets. AB - Six multicatheterized beef steers (421 +/- 21 kg BW) were used to predict the effect of dietary concentrate level on blood flow and net flux of urea and other metabolites across splanchnic tissues. Diets ranged from 0% (switchgrass hay) to 90% concentrate (10% switchgrass hay, 89% cracked corn, 1% urea). Daily DMI varied from 8.01 to 5.34 kg/d. Nitrogen intake (99 g/d) and calculated ME intake (16.8 Mcal/d) were equal among diets. As dietary concentrate increased from 0 to 90%, liver blood flow decreased from 850 to 795 L/h, portal-drained visceral (PDV) blood flow decreased from 750 to 620 L/h, and mesenteric-drained visceral (MDV) blood flow decreased from 270 to 250 L/h. Liver release of urea N was 94 mmol/h when dietary concentrate was less than 20%, then increased to 146 mmol/h at 55% concentrate. Urinary excretion of urea N was 13 mmol/h or less when dietary concentrate was 20% or less, increased to 53 mmol/h at 55% concentrate, then continued to increase to 76 mmol/h at 90% concentrate. Transfer of urea N to PDV ranged from 71 to 91 mmol/h and transfer to MDV ranged from 0 to 10 mmol/h among diets. As dietary concentrate increased from 27 to 63%, VFA release by PDV decreased, net MDV and splanchnic release of glucose increased, and splanchnic tissues switched from net uptake to net release of L-lactate. Net PDV release or liver removal of ammonia or alpha-amino N and net liver release of glucose were not affected. We conclude that the liver responded to changes in the percentage of dietary concentrate by altering urea production and by altering the role of lactate in intermediary metabolism. PMID- 8728015 TI - Testing for undesirable traits in cattle: an Australian perspective. AB - A variety of autosomal recessive defects, many lethal to the newborn calf, have been recognized in Australia. Definition of a defect at the biochemical or molecular level facilitates development of heterozygote detection tests essential for efficient disease control programs. The prevalence of alpha-mannosidosis in Angus and Murray Greys, generalized glycogenosis in Brahmans and Shorthorns, and citrullinemia in Holstein/Friesians has been reduced as a result of industry sponsored disease-control programs. These defects were disseminated as a consequence of selection focused on desirable traits carried by individuals. In the long term, an increase in crossbreeding in commercial beef production will reduce the significance of recessive defects. Caution will be required to reduce the risk of dissemination of recessive defects resulting from increased selection pressure within the dairy industry presently dominated by Holstein/Friesians. PMID- 8728016 TI - Changes of fatty acid composition and oxidative metabolism of human neutrophils migrating into an inflammatory exudate. AB - The peripheral blood neutrophils and the neutrophils accumulated into a skin window experimental exudate were compared for their ability to release superoxide anion (O2-) and for their fatty acid composition, determined by capillary gaschromatography. The basal O2- release and the phorbol myristate acetate (PMA) induced O2- release were not significantly different in the two neutrophil populations, while in response to formyl-methionyl-leucyl-phenylalanine (fMLP) the exudate cells showed an activity that was two fold higher than that of blood cells. The most significant changes of fatty acid composition of exudate versus blood cells were the following: i) increase of C16:0 (palmitic acid) from 21.3 +/ 1.2% to 23.5 +/- 1.3% (+ 10.2%) of total fatty acids (p < 0.001),ii) increase of C18:2 (linoleic acid) from 9.3 +/- 1.7% to 11.0 +/- 2.1% (+ 18.3%) (p = 0.005), iii) decrease of C20:4 (arachidonic acid) from 12.8 +/- 1.6% to 10.6 +/- 1.2% ( 17.3%) (p < 0.001), whereas C18:1 (oleic acid) did not change. The total content of saturated or unsaturated fatty acids did not change. In exudate cells, a strong negative correlation was found between palmitic acid content and O2- release in response to both fMLP and PMA (r = -0.52, p < 0.02 and r = -0.49, p < 0.05, respectively) whereas arachidonic acid correlated positively, but weakly, with O2- (r = 0.40, p = 0.07 and r = 0.47, p = 0.05, with fMLP and PMA as stimulants respectively). A positive correlation was also found between the arachidonic acid content of blood cells and the number of cells that migrated into the inflammatory exudate. These results indicate that the process of extravasation from blood into the exudate causes specific and consistent modifications of the fatty acid composition of neutrophils and suggest that these modifications have a role in the activation and the regulation of the O2- forming system. PMID- 8728017 TI - Human neutrophil NADPH-oxidase activity is inhibited by lazaroids. PMID- 8728018 TI - The direct effect of interferon-gamma on human eosinophilic leukemia cell lines: the induction of interleukin-5 mRNA and the presence of an interferon-gamma receptor. AB - The EoL-1 and EoL-3, human eosinophilic leukemia cell lines, have been used as models for studying the maturation and the function of human eosinophils. We investigated the effects of interferon-gamma (IFN-gamma) on superoxide anion (O2 ) production of these cell lines and interleukin-5 (IL-5) mRNA expression in the EoL-1. O2- was measured by chemiluminescence of MCLA, one of cypridina luciferin analogs. The O2- production of fMLP-stimulated EoL-1 and EoL-3 was increased by the IFN-gamma treatment. IL-5 mRNA expression was detected in the IFN-gamma treated EoL-1 by reverse transcription-polymerase chain reaction (RT-PCR). Further, we examined IFN-gamma receptor 1 mRNA expression in these cell lines and peripheral blood eosinophils by means of northern blot hybridization. IFN-gamma receptor 1 mRNA was detected in the EoL-3 and the IFN-gamma-treated EoL-1. A weak expression of IFN-gamma receptor 1 mRNA was detected in peripheral blood eosinophils isolated from a patient with eosinophilia. These results suggest that IFN-gamma may act on eosinophils directly through its receptor. PMID- 8728019 TI - The intratracheal administration of endotoxin: X. Dexamethasone downregulates neutrophil emigration and cytokine expression in vivo. AB - Intratracheal instillation of endotoxin (LPS) causes acute pulmonary inflammation characterized by the accumulation of plasma proteins and leukocytes within the pulmonary airways. The synthetic glucocorticoid dexamethasone 1) inhibits the LPS initiated vascular leak of plasma proteins into the airspace, 2) inhibits the LPS initiated emigration of neutrophils and lymphocytes into the airspace in a dose dependent fashion, and 3) inhibits LPS-initiated mRNA and/or bronchoalveolar lavage protein expression of cytokines (TNF, IL-1 and IL-6) and chemokines (MIP-1 alpha, MIP-2 and MCP-1). In conclusion, dexamethasone inhibits both the vascular and cellular aspects of acute inflammation by downregulation of a broad spectrum of inflammatory cytokines and chemokines. PMID- 8728020 TI - Bacterial lipopolysaccharide-stimulated nitric oxide generation is unrelated to concurrent cytotoxicity of bovine alveolar macrophages. AB - Nitric Oxide (NO) is a reactive metabolite produced by stimulated macrophages, and it has been demonstrated to exert cytotoxic actions on a number of microbes, parasites, and tumor cells. In addition, NO has been reported to have an autotoxic effect on murine macrophages, its site of synthesis. We have investigated the relationship of NO generation to cytotoxicity of bovine alveolar macrophages (AM) in vitro, and have also assessed the effects of several modulators of cellular function on this relationship. NO was generated in cultures of AM using sodium nitroprusside (SNP) and measured as [nitrite]. Cellular viability of AM reflected a strong, negative correlation with the concentration of NO/nitrite in supernatants (r = -0.987). Supernatants with nitrite concentrations in excess of 30 microMs were correlated with cytotoxicity. AM stimulated with the potent combination of endotoxin (Lipopolysaccharide, LPS; 10 ng/ml) and recombinant bovine IFN gamma (100 U/ml) also exhibited cytotoxicity over a 48-hour incubation period, and cells deteriorated to an average viability of 72.3% as compared to unstimulated control macrophages. In some cases the viability of macrophages was much lower. Even though LPS-mediated cytotoxicity occurred, the [nitrite] produced in supernatants during the 48-hour period (12.23 microMs) was well below the minimum concentration of SNP-generated NO required to induce cytotoxicity to macrophages. N(G)-monomethyl-L-arginine (N(G)MMA, 2 mM) is a competitive inhibitor of NO synthesis and was found to reduce nitrite concentrations from 12.23 microMs to 1.56 microMs in supernatants of LPS stimulated AM, but this reduction did not promote increased viability of AM. Other modulators of cellular function including phenylbutazone (PBZ, 100 microMs), flunixin meglumine (FM, 100 microMs) and interleukin-4 (IL-4, 100 ng/ml) modestly inhibited synthesis of NO, but did not improve cellular viability. These results suggest that relatively high concentrations of exogenously-generated NO are toxic to AM in vitro, but the quantity of endogenously-generated NO synthesized by LPS-stimulated bovine AM is usually below the threshold for toxicity. Cytotoxicity occurs independently of NO synthesis, and factors other than NO are apparently responsible for LPS-related cytotoxicity to bovine macrophages. PMID- 8728021 TI - Effects of insulin, dexamethasone and cytokines on alpha 1-acid glycoprotein gene expression in primary cultures of normal rat hepatocytes. AB - While the effects of insulin, dexamethasone and cytokines on alpha 1-acid glycoprotein gene expression have been investigated in various hepatoma cell lines, the individual and combined effects of these components on the expression of this gene have been rarely studied in cultured normal rat hepatocytes. In this cell model, we have shown that mRNA levels of alpha 1-acid glycoprotein were not decreased at least during the first 24 h of culture under basal conditions. During these short-term cultures, the expression of alpha 1-acid glycoprotein in normal hepatocytes showed a high degree of responsiveness to dexamethasone alone (20-fold increase) and to dexamethasone associated with various cytokines (interleukin-1 beta, interleukin-6 and tumor necrosis factor alpha) with a 40 to 100-fold increase depending on the cytokine. Insulin alone did not modify alpha 1 acid glycoprotein mRNA; however, this hormone exerted a positive effect (about 50% increase) in the presence of dexamethasone or dexamethasone with cytokines. These results indicate that the regulation of alpha 1-acid glycoprotein in cultured normal rat hepatocytes presents major differences when compared to reported observations in rat hepatoma cell lines. PMID- 8728023 TI - Synergistic cooperation between thapsigargin and phorbol ester for induction of nitric oxide synthesis in murine peritoneal macrophages. AB - The biochemical transductional events involved in NO synthesis are not fully understood. These studies, therefore, were undertaken to elucidate the role of intracellular calcium and protein kinase C (PKC) in the induction of nitric oxide (NO) synthesis in murine peritoneal macrophages. Thapsigargin (TG), Ca(2+)-ATPase inhibitor of endoplasmic reticulum, had modest activity on NO synthesis by itself, whereas phorbol ester, PKC activator, alone had no effect. When TG was used in combination with phorbol ester, there was a marked cooperative induction of NO synthesis in a dose-dependent manner. The optimal effect of phorbol ester was shown in the first 6 h after TG treatment. In addition, the ability of TG with phorbol ester on NO synthesis could be mimicked by another chemically unrelated inhibitor of Ca(2+)-ATPase, 2,5-di-(t-butyl)-1, 4-benzohydroquinone, and Ca2+ ionophore, A23187. This increase of NO synthesis was reflected as increased amount of NO synthase (NOS) mRNA, as determined by Northern blotting. Intracellular Ca2+ transient by TG was not affected in the presence or absence of extracellular Ca2+, indicating that TG must be effective on cytosolic Ca2+ pool. In addition, chelation of intracellular Ca2+ by acetoxymethyl ester of 1,2-bis-(2 aminophenoxy)-ethane-N,N,N',N'-tetraacetic acid (BAPTA/AM), an intracellular Ca2+ chelating agent, blocked TG- or TG + PMA-induced NO production. PKC inhibitors such as staurosporine or polymyxin B reduced only the synergistic cooperative effect of TG with phorbol ester without affecting TG-induced NO production. In addition, when the cells were pretreated with phorbol ester before TG treatment, there was no synergy between TG and phorbol ester, indicating that PKC is not directly involved in the expression of NOS but involved in "triggering" signal. Secretion of NO corresponded with tumor cell killing, but TG plus phorbol ester activated macrophages failed to kill tumor cell targets in the presence of Ng monomethyl-L-arginine. Collectively, these data illustrate that mobilization of intracellular Ca2+ provides a "priming" signal for induction of NOS gene expression by itself and it also requires PKC as a "triggering" signal for macrophage tumoricidal activity. PMID- 8728022 TI - Synergistic effects of thalidomide and poly (ADP-ribose) polymerase inhibition on type II collagen-induced arthritis in mice. AB - The present study investigates synergistic effects of the TNF-alpha inhibitor thalidomide and the poly(ADP-ribose) polymerase (PARP)-inhibitor nicotinic acid amide (NAA) in male DBA/1 hybird mice suffering from type II collagen-induced arthritis. Parameters including the arthritis index, chemiluminescence and anti collagen antibody titers were used for the assessment of disease activity: The disease courses demonstrated clearly an inhibitory effect of thalidomide. NAA inhibited established collagen arthritis in a dose-dependent manner. The combined application of thalidomide and NAA caused a powerful synergistic inhibition of arthritis. Furthermore, thalidomide and NAA were tested ex vivo for their inhibition of the NADPH oxidase-dependent generation of reactive oxygen species by activated neutrophils and monocytes in unseparated human blood. Our data show that type II collagen-induced arthritis can be suppressed by the simultaneous inhibition of TNF-alpha, PARP, and NADPH oxidase. PMID- 8728024 TI - Measurement of hydroxyl radicals catalyzed in the immediate vicinity of DNA by metal-bleomycin complexes. AB - A recently developed sensitive fluorimetric assay has been used to examine whether free hydroxyl radicals (HO.) are generated in the immediate vicinity of DNA by Fe(II)-bleomycin. When aqueous solutions of SECCA (the succinimidyl ester of coumarin-3-carboxylic acid) are irradiated with gamma rays or incubated with Fe(II)-bleomycin or Fe (II)-EDTA in the presence of ascorbate and H2O2, 7-hydroxy SECCA, a fluorescent product of the interaction of HO. with SECCA, is generated. Studies with catalase and several HO. scavengers indicate that the fluorescence induction is mediated by HO. On the contrary, Cu(II)-bleomycin complexes under similar conditions fail to induce 7-hydroxy-SECCA fluorescence. When SECCA is conjugated to DNA via SECCA-polylysine-DNA complexes and incubated in the same iron-containing systems, the relative ability of the scavengers to reduce the fluorescence again demonstrates the generation of 7-hydroxy-SECCA by HO. However, while the fluorescence is practically eliminated by high concentrations of DMSO (100 mumols dm-3) in the systems with Fe(II) or Fe(II)-EDTA, it is not possible to reduce it similarly in the case of Fe(II)- bleomycin. These data demonstrate the generation of HO. by Fe(II)-bleomycin in the immediate vicinity of DNA. Because the experiments simulate the lifetime of HO. expected in cells, these data suggest that, if such DNA-associated HO. radicals are also produced in vivo by bleomycin, these would not be scavengable by intracellular scavengers and they could interact with chromatin. PMID- 8728025 TI - Isoluminol-enhanced chemiluminescence: a sensitive method to study the release of superoxide anion from human neutrophils. AB - A very sensitive chemiluminescence (CL) method was developed for the determination of respiratory burst products generated by the NADPH-oxidase in human neutrophils. Despite the fact that the CL reaction is peroxidase dependent, hydrogen peroxide was found not to participate in the light generating reaction. Phagocytic cells were mixed with isoluminol, a chemiluminescence substrate that detects extracellularly released oxygen species only. Owing to the fact that the availability of released cellular myeloperoxidase is a limiting factor in the reaction, extra peroxidase (horseradish peroxidase; HRP) has to be added to the measuring system. From the fact that the response to phorbol myristate acetate (PMA), as well as to formyl-methionyl-leucyl-phenylalanine (fMLP) was almost totally inhibited by superoxide dismutase (SOD), we conclude that O2.- is the reacting oxygen species measured. The assay system described is well suited for real-time studies of superoxide anion release from activated neutrophils. With the technique, the release of O2.- can be detected from as few as 250 neutrophils. PMID- 8728026 TI - Noxious effects of oxygen reactive species on energy-coupling processes in Ehrlich ascites tumor mitochondria and the protection by pantothenic acid. AB - Irradiation of Ehrlich ascites tumor cells with ultraviolet light or exposure to the Fenton reaction results in lesions in the mitochondrial energy-coupling system. Formation of the membrane potential and its utilization for ATP synthesis are more affected than the respiratory chain. Preincubation of the cells with pantothenic acid or its derivatives which can serve as precursors of CoA largely protects against the damage of mitochondrial energetics by oxygen reactive species formed by UV light or the Fenton reaction. Incubation of Ehrlich ascites tumor cells with pantothenic acid increases their content of glutathione (most of which is present in the reduced form) by 40%. It is concluded that the protective effect of precursors of CoA against lesions of the mitochondrial energy-coupling system by oxygen reactive species is mainly due to removal of free radicals and peroxides by glutathione peroxidase and phospholipid hydroperoxide glutathione peroxidase. PMID- 8728027 TI - Effect of natural antioxidant tanshinone II-A on DNA damage by lipid peroxidation in liver cells. AB - Tanshinone II-A (TSII-A) isolated from the root of Salvia miltorrhiza Bunge, a traditional medicine in China, is a derivative of phenanthrenequinone, which is known to have antioxidant properties. In the present study, effects of TSII-A on DNA damage by lipid peroxidation were investigated using liver cells, labeled with [3H] arachidonic acid, in the presence of FeCl2-DTPA. The results show that the nuclear DNA isolated from treated cells had higher radioactivity compared to controls and the radioactivity increased with longer incubation times. Purified lipid-DNA adducts had a characteristic fluorescent spectra and showed a decrease of hyperchromicity and melting point. TSII-A could inhibit the association of peroxidation products with DNA in liver cells and prevent a decrease in cell viability and in the the activity of O6-methylguanine acceptor protein with increasing incubation time. Compared with other antioxidants, TSII-A had a higher inhibitory ratio, which was similar to vitamin E and butylated hydroxy-toluene (BHT), but markedly stronger than NaN3, mannatol, and superoxide dismutase (SOD). These data suggest that TSII-A represents a new and effective antioxidant that inhibits the association of lipid peroxidation products with DNA. Its protective effect may be through breaking the chain reactions of peroxidation by scavenging lipid free radicals, thereby decreasing their cytotoxicity. PMID- 8728028 TI - Horseradish peroxidase-catalyzed sulfoxidation of promethazine and properties of promethazine sulfoxide. AB - Promethazine sulfoxide was obtained with a quantitative yield in a horse radish peroxidase-catalyzed reaction of promethazine and hydrogen peroxide and was also prepared by direct chemical synthesis. The enzymatic sulfoxidation of promethazine was studied in vitro as a function of pH, promethazine, and hydrogen peroxide concentration. Promethazine sulfoxide inhibits with an apparent K(i) of 59.7 microM at pH 5.5 the enzymatic reaction, followed spectrophotometrically, polarographically, potentiometrically, and luminometrically. The reaction was also inhibited by ascorbic acid (K(i) 26.8 microM) and glutathione (K(i) 41.8 microM). The spectrophotometric techniques employed, together with ESR spectrometry, allowed the identification of at least three radical species formed in the course of the reaction. Promethazine sulfoxide is devoid of the antioxidant effect exhibited by promethazine on rat brain synaptosomes. The sulfoxide also lacks photosensitizing action, while retaining the neuroleptic effect of the parent compound. PMID- 8728029 TI - Activation of the HIV long terminal repeat and viral production by H2O2-vanadate. AB - The long terminal repeat (LTR) of human immunodeficiency virus type 1 (HIV-1) contains sequences required for the initiation of gene transcription. Among the substances known to activate the HIV-1 LTR is hydrogen peroxide (H2O2). We report here that H2O2-induced activation of the LTR in the macrophage cell line THP-1 and the lymphocyte cell line, Jurkat, is greatly increased by vanadate. Activation of the LTR by phorbol myristate acetate, tumor necrosis factor alpha, lipopolysaccharide, or Staphylococcus epidermidis extract was not increased by vanadate, indicating some selectivity for H2O2. H2O2 and vanadate also acted synergistically to increase the production of HIV-1 virions by the latently infected macrophage cell line U-1 as determined by p24 antigen release and the detection of intact virions by electron microscopy. Effects were observed at H2O2 and vanadate concentrations down to 3 x 10(-6) M, with high concentrations leading to cell toxicity. Catalase was strongly inhibitory when added prior to the interaction of H2O2 and vanadate, but was considerably less inhibitory when the H2O2 and vanadate were allowed to preincubate prior to the catalase addition. H2O2 reacts with vanadate to form peroxides of vanadate that have potent biological effects. Our findings suggest that among these is the activation of the HIV-1 LTR. PMID- 8728030 TI - Age differences in sensitivity to H2O2- or NO-induced reductions in K(+)-evoked dopamine release from superfused striatal slices: reversals by PBN or Trolox. AB - Previous research has indicated that many age-related functional alterations may be the result of a decreased ability of the organism to respond to oxidative stress (OS). However, this hypothesis is based on indirect indices of function (e.g., increased vulnerability of hepatocytes from senescent animals to H2O2 induced DNA damage, increases in lipofuscin accumulation). More direct tests of this hypothesis, especially as it relates to brain aging, have not been extensively undertaken. Present experiments were carried out to make such tests by examining age differences in the sensitivity to OS on reductions in striatal dopamine (DA) release. Thus, K(+)-evoked DA (K(+)-ERDA) release from superfused striatal slices from young (6-8 month) and old (24-25 month) animals was examined following either: (a) application of the NO-generator sodium nitroprusside or (b) preincubation with H2O2. In order to assess the specific effects of OS on muscarinic (mAChR) sensitivity, oxotremorine-enhancement of K(+) -ERDA was examined following incubation with H2O2. Results showed that the striatal tissue from the old animals showed greater sensitivity to both H2O2 and NO than young animals, and stimulated DA decreased at lower concentrations of these agents (e.g., NO--100 microM young, 30 microM old). In addition, H2O2 was also effective in reducing oxo-enhanced K(+)-ERDA and was more effective as a function of age. If the striatal tissue was incubated in either Trolox (alpha-tocopherol) or alpha phenyl-n-tert-butyl nitrone (PBN) prior to OS, the negative effects of NO. and H2O2 were reversed in both age groups. Results are discussed in terms of age related membrane and endogenous antioxidant alterations that could induce increases in sensitivity to OS and the specificity of antioxidants in reducing this sensitivity in key functional systems. PMID- 8728031 TI - Radicals generated by bone cutting and fracture. AB - When bone fractures, a remarkably high yield of radicals is generated. By working at 77K, these are stable enough for detection and study. Major species have been identified by ESR (EPR) spectroscopy as alkyl peroxyl radicals and RCH2. radicals. Also, a species thought to be O.- radicals, formed in the mineral phase, was detected in relatively low yield. It is suggested that as a break is generated, the mineral crystallites separate at grain boundaries with no major chemical damage, but that the tightly bound collagen strands running through the mineral phase are forced to break homolytically. Some probably react together, others react with oxygen, while the remainder are trapped in their primary form. The characteristic hyperfine triplet is assigned in part to RCH2. radicals from glycine units, which constitute about one-third of the total amino acid components in collagen, and in part to RNHCO radicals. The possible significance of the formation of active radicals during bone fracture is discussed. PMID- 8728032 TI - Oxidation of spin-traps by chlorine dioxide (ClO2) radical in aqueous solutions: first ESR evidence of formation of new nitroxide radicals. AB - The reactivities of the chlorine dioxide (ClO2), which is a stable free radical towards some water-soluble spin-traps were investigated in aqueous solutions by an electron spin resonance (ESR) spectroscopy. The ClO2 radical was generated from the redox reaction of Ti3+ with potassium chlorate (KClO3) in aqueous solutions. When one of the spin-traps, 5,5-dimethyl-1-pyrroline N-oxide (DMPO), was included in the Ti3+-KClO3 reaction system, ESR spectrum due to the ClO2 radical completely disappeared and a new ESR spectrum [aN(1) = 0.72 mT, aH(2) = 0.41 mT], which is different from that of DMPO-ClO2 adduct, was observed. The ESR parameters of this new ESR signal was identical to those of 5,5 dimethylpyrrolidone-(2)-oxyl-(1) (DMPOX), suggesting the radical species giving the new ESR spectrum is assignable to DMPOX. The similar ESR spectrum consisting of a triplet [aN(1) = 0.69 mT] was observed when the derivative of DMPO, 3,3,5,5 tetramethyl-1-pyrroline N-oxide (M4PO) was included in the Ti3+-KClO3 reaction system. This radical species is attributed to the oxidation product of M4PO, 3,3,5,5-tetramethylpyrrolidone-(2)-oxyl-(1) (M4POX). When another nitrone spin trap, alpha-(4-pyridyl-1-oxide)-N-t-butylnitrone (POBN) was used as a spin-trap, the ESR signal intensity due to the ClO2 radical decreased and a new ESR signal consisting of a triplet [aN(1) = 0.76 mT] was observed. The similar ESR spectrum was observed when N-t-butyl-alpha- nitrone (PBN) was used as a spin-trap. This ESR parameter [a(N)(1) = 0.85 mT] was identical to the oxidation product of PBN, PBNX. Thus, the new ESR signal observed from POBN may be assigned to the oxidation product of POBN, POBNX. These results suggest that the ClO2, radical does not form the stable spin adducts with nitrone spin-traps, but oxidizes these spin-traps to give the corresponding nitroxyl radicals. On the other hand, nitroso spin-traps, 5,5-dibromo-4-nitrosobenzenesulfonate (DBNBS), and 2-methyl-2 nitrosopropane (MNP) did not trap the ClO2 radical. This result indicates that an unpaired electron of the ClO2 radical is localized on oxygen atom, because nitroso spin-traps cannot form the stable spin adduct with oxygen-centered radical. PMID- 8728033 TI - Can superoxide organic chemistry be observed within the liposomal bilayer? AB - --2-(Dimethylamino) fluorene (1a) and 5-benzoyloxy-2,3,7,8,12,13,17,18 octaethylporphyrin (4) react with superoxide anion radical (generated from KO2/18 crown-6 polyether) in aprotic media. Yet, when incorporated into the lipid bilayer of dimyristoyl phosphatidylcholine liposomes, these two substrates are inert to superoxide, generated enzymatically (xanthine oxidase/acetaldehyde) or radiolytically (60Co or 137Cs source/formate solution). On the other hand, 7 acetoxy-4-methylcoumarin (6), which reacts with superoxide in aprotic media yielding the corresponding 4-methylumbelliferone (7), also gives the same product when incorporated within the liposomal bilayer and reacted with radiolytically or enzymatically generated superoxide. In the latter case, the reaction is inhibited by SOD. NMR studies indicate that in contradistinction to the highly lipophilic 1a and 4, which presumably lie well within the lipid bilayer, 7 lies in a highly polar region of the bilayer. These results suggest that superoxide anion does not penetrate deep into the liposomal bilayer; nevertheless, superoxide reactions can, indeed, be observed, provided the active site of the substrate lies at or near the lipid-water interface. PMID- 8728034 TI - A study of the synergistic interaction of asbestos fibers with cigarette tar extracts for the generation of hydroxyl radicals in aqueous buffer solution. AB - Several models attempt to explain the synergistic increase in lung cancer among workers exposed to asbestos fibers, who were smokers at the same time. It is known that reactive oxygen species (ROS) are important mediators in asbestos induced diseases, especially cancer. We studied quantitatively the formation of ROS (by spin trapping with DMPO) in aqueous buffer suspensions containing crocidolite (UICC), chrysotile (UICC and commercial, long fibers) alone, and in combination with aqueous cigarette tar extracts. It was observed that asbestos and cigarette tar act in a cooperative or synergistic way in the generation of hydroxyl radical spin adducts. Grinding of asbestos fibers and addition of EDTA (iron chelator) enhanced the intensity of the ESR signal. This enhancement progressed with time, probably due to the reaction of the extracted iron with the slow released hydrogen peroxide from tar extracts. It was observed a fivefold increase in the ESR signal (for crocidolite and aqueous tar extracts) in the formation of hydroxyl radicals via an iron-catalyzed Fenton reaction. These experimental results are suggest to be strong evidence to the fact that lung cancer has been found in asbestos workers exposed to high concentrations of fibers in the working environment who were smokers, and only rarely in nonsmokers. PMID- 8728035 TI - Association between 8-hydroxy-2'-deoxyguanosine formation and DNA strand breaks mediated by copper and iron. AB - Oxidative DNA damage is involved in diverse biological phenomena and consists of several kinds of lesions, mainly, strand breaks, base modifications, and DNA protein crosslinking. However, little is known about the existence of a chemical relationship among them or the ratio by which these different types of lesions are produced. In the present study we investigated whether a relationship exists between DNA strand breakage and base modification. We selected cupric [Cu(II)] and ferric [Fe(III)] ions for this study because these transition metals are active catalysts of DNA damage in vivo. Supercoiled plasmid DNA pZ189 was treated with Cu(II) or Fe(III) in the presence of different reducing agents. We measured in each sample both the number of DNA single-strand breaks (SSB) by quantitative electrophoresis and the amount of a modified DNA base, 8-hydroxy-2' deoxyguanosine (8-OHdG) by HPLC with simultaneous electrochemical (EC) and spectrophotometric detection. The number of DNA SSBs produced was linearly related to the number of 8-OHdG present. The regression of the number of SSBs as a function of the number of 8-OHdG is expressed by the equation [SSBs] = b x [8 OHdG], where b = 1.7, 2.0, 2.7, 1.7, and 9.4, for Cu(II) in the presence of H2O2, L-cysteine and L-ascorbate, and for Fe(III) in the presence of H2O2 and L ascorbate, respectively. The linear correlation we observed between the production of SSB and 8-OHdG mediated by Fe(III) and by Cu(II) suggests that these products may arise via a common chemical mechanism and could allow an easier and more precise estimation of DNA breakage. PMID- 8728036 TI - Lung poly(ADP-ribose) and NAD+ concentrations during hyperoxia and niacin deficiency in the Fischer-344 rat. AB - Sublethal exposure to hyperoxia in vivo induces oxidative damage that leads to destruction of the pulmonary endothelium, pleural effusion, and eventual pulmonary fibrosis. DNA is a potential target for reactive oxygen species in this system; the principle types of damage to DNA during hyperoxia are single-strand breaks and oxidant damage to bases. Poly(ADP-ribosyl)ation, a posttranslational modification of nuclear proteins, is stimulated by strand breaks in DNA and is required for effective repair of many types of DNA lesions. In this study we have measured lung tissue NAD+ and poly(ADP-ribose) concentrations in response to hyperoxia and niacin deficiency in rats. Male weaning Fischer-344 rats consumed niacin-deficient (ND) or niacin-replete pair-fed (PF) diets for 7 d. Rats from each diet group (n = 6) were then housed in normobaric 85% oxygen for 5 d. Normoxic controls were maintained in air. Hyperoxia increased lung poly(ADP ribose) concentration by 35% in PF rats, but did not significantly increase levels in ND rats. Niacin deficiency decreased lung NAD+ in normoxic rats, but surprisingly, this deficit was partially reversed by hyperoxia. Liver NAD+ levels increased by 21% during hyperoxia in both diet groups. Heart and kidney NAD+ were unaffected by hyperoxia. Blood was the only tissue measured in which NAD+ was decreased by hyperoxia. Dietary treatment did not affect the increase in the lung wt/b. wt. ratio resulting from hyperoxia. This is the first report in the literature of lung tissue poly(ADP-ribose) measurement. Results show that hyperoxia causes a marked increase in lung poly (ADP-ribose) concentration, but also suggest an adaptation of whole-animal NAD+ metabolism to hyperoxia during niacin deficiency. PMID- 8728037 TI - Detection of JC virus DNA in the peripheral blood leukocytes of HIV-infected patients. AB - OBJECTIVES: To assess whether JC virus (JCV) DNA is frequently harboured by peripheral blood leukocytes (PBL) in HIV-positive patients, before the onset of progressive multifocal leukoencephalopathy (PML). DESIGN: The polyomavirus JCV induces PML in immunocompromised persons and particularly AIDS patients. Leukocytes may play a central part in the onset of PML, but their precise role in JCV latency and reactivation still remains hypothetical. The controversial presence of JCV DNA in PBL has been, until now, investigated only among small groups of patients. We therefore studied 157 HIV-positive persons and compared them with 65 HIV-negative immunocompromised patients. METHODS: DNA was extracted from PBL. The presence of JCV DNA was demonstrated by the polymerase chain reaction (PCR) alone or combined with a molecular hybridization assay. RESULTS. The presence of JCV DNA was ascertained by PCR and hybridization in 28.9% of 135 HIV-infected persons at all stages of HIV infection and only 16.4% of 61 HIV negative immunocompromised patients. No correlation could be drawn between the detection of JCV DNA and the clinical or biological status of the HIV-positive patients. CONCLUSIONS: JCV DNA is detectable in the PBL of 28.9% of HIV-infected persons, even in the early stages of infection. JCV is more seldomly amplified in HIV-negative immunocompromised patients. Further work is in progress to determine the prognostic value of the presence of JCV DNA in the blood of HIV-positive patients. PMID- 8728038 TI - Rhodococcus equi infection in HIV-infected patients. AB - OBJECTIVE: To report clinical and microbiological features and response to treatment in HIV patients with Rhodococcus equi infection. DESIGN: Retrospective study. SETTING: Inpatients admitted to two Infectious Diseases Departments in a community-based hospital. PATIENTS: A total of 12 HIV-positive patients with R. equi infection. MAIN OUTCOME MEASURES: Clinical status, radiological finding, microbiological, haematochemical and immunological tests, and response to treatment. RESULTS: Twelve patients (11 men, six injecting drug users) were diagnosed with R. equi infection. Fever and cough were the principal clinical signs on presentation. Mean CD4+ count at the time of diagnosis was 47.67 x 10(6)/l (SD, 49.2 x 10(6)/l). In 58.3% of the cases the diagnosis of R. equi infection followed the appearance of an AIDS-defining illness. The most frequent radiological findings were cavitary lesions (41.7%) and lung consolidation (33.3%). In 83% of cases, R. equi was isolated from blood and in 33.3% cases from sputum. Test of chemosensitivity showed sensitivity to vancomycin (100%), teicoplanin (100%), ceftriaxone (80%), erythromycin (71%) and ciprofloxacin (66%). Clinical response alone with the disappearance of the presenting signs was observed in nine of the 12 cases (75%); complete response was observed in two cases. Seven patients died with a mortality rate of 58.3% and a mean survival of 5.75 months (SD, 6.48 x 10(6)/l). CONCLUSIONS: R. equi should be considered in the differential diagnosis of pulmonary of disseminated infections in patients with HIV infection. Blood culture may be the most sensitive means of diagnosis. Other studies are needed to determine the most effective choice and duration of antibiotic therapy. PMID- 8728039 TI - Retention of HIV-1 inside infected MOLT-4 cells in association with adhesion induced cytoskeleton reorganization. AB - OBJECTIVE: To study the mechanism of the suppression of HIV release during cell to-cell adhesion. DESIGN AND METHODS: To investigate the effects of cell-to-cell adhesion on HIV release in association with cytoskeletal elements, chronically HIV-infected T cells were cocultured with different adherent cell lines, cultured on a fibronectin-coated surface, or treated with cytochalasin D. The amount of viral protein released in the culture supernatant and retained inside the cells was monitored by a p24 enzyme-linked immunosorbent assay and Western blotting. For F-actin staining, cells were stained with FITC-labelled phalloidine and examined by immunofluorescence microscopy. RESULTS: Cocultivation resulted in a reduced amount of virus in the culture supernatant and induced the retention of viral protein inside the infected cells. On adhesion to cells, the F-actin of the infected cells was polarized towards the cell periphery from a diffuse cytoplasmic distribution. Similar data were obtained when the infected cells were treated with cytochalasin D or adhered to fibronectin. CONCLUSION: Cell-to-cell adhesion induced polarization of F-actin, which might facilitate HIV retention inside infected T cells. PMID- 8728040 TI - Differential activation of the extracellular signal-regulated kinase, Jun kinase and Janus kinase-Stat pathways by oncostatin M and basic fibroblast growth factor in AIDS-derived Kaposi's sarcoma cells. AB - OBJECTIVES: To determine the integration of signalling pathways associated with two recognized Kaposi's sarcoma (KS) growth factors, oncostatin M (OSM) and basic fibroblast growth factor (bFGF), in the induction of KS cell proliferation. DESIGN AND METHODS: We used protein kinase assays, protein-DNA interactions and AP-1 luciferase assays to study the extracellular signal-regulated kinase (ERK), Janus kinase (JAK)-Stat and Jun kinase (JNK) pathways in AIDS-derived KS cells during stimulation with OSM and bFGF. RESULTS: Treatment with OSM-induced activation of receptor-associated JAK and phosphorylation of Stat1 and Stat3. Stat1/Stat3 heterodimers interacted with known gamma-interferon-activated sites like elements such as the sis-inducible element (SIE) in the C-fos promoter. In contrast, ligation of the bFGF receptor induced Stat3 phosphorylation and its association with the bFGF receptor, but failed to induce JAK activity or protein complexes which interact with GAS-like oligonucleotides. OSM also induced the activation of ERK2 by activating the serine/threonine kinases Raf-1 and [mitogenactivated protein kinase (MAPK) ERK kinase (MEK1)]-1, while bFGF failed to activate any of the above components. Both OSM and bFGF activated the JNK pathway, along with the activation of MEKkinase (MEKK)-1. JNK control the transcriptional activation of c-Jun. Because the above pathways exert an effect on the expression or activation of activation protein (AP)-1 components, we confirm that OSM and bFGF induce TPA response element (TRE)-luciferase activity synergistically. CONCLUSION: We demonstrate that OSM and bFGF activate distinct as well as shared signalling cascades in KS cells, which integrate to provide a synergistic AP-1 response by which OSM and bFGF may sustain KS cell growth. PMID- 8728041 TI - Efficacy of 2-month total parenteral nutrition in AIDS patients: a controlled randomized prospective trial. The French Multicenter Total Parenteral Nutrition Cooperative Group Study. AB - OBJECTIVE: To evaluate the efficacy of total parenteral nutrition in AIDS patients. DESIGN: A prospective, randomized, controlled, multicentre trial. METHODS: Over a period of 2 months, 31 malnourished and severely immunodepressed AIDS patients were assigned to receive either dietary counselling (n = 15) or home total parenteral nutrition (TPN; n = 16) via a central venous access after an educational program. Results were analysed by intent-to-treat basis. RESULTS: Bodyweight change was +8 kg (+13 +/- 3%) in the TPN group and -3 kg (-6 +/- 2%) in the control group (P < 0.0006). Lean body mass increased in the TPN group (+9 +/- 3%) and decreased in the control group (-5 +/- 3%; P < 0.004) while body cell mass increased in the former (+15 +/- 4%) and decreased in the latter (-12 +/- 6%; P < 0.002). Nutritional subjective global assessment, subjective self reported health feeling and Karnofsky index were also improved by TPN. Infection line sepsis incidence remained low (0.26 per 100 patient-days). However, no difference in survival rate was exhibited between the two groups by the log-rank test. CONCLUSION: We conclude that home TPN is an efficient treatment of malnutrition in severely immunodepressed AIDS patients. PMID- 8728042 TI - The diffuse infiltrative lymphocytosis syndrome. Clinical and immunogenetic features in 35 patients. AB - OBJECTIVE: To study the epidemiological, clinical, serological and immunogenetic features of the diffuse infiltrative lymphocytosis syndrome (DILS). DESIGN: Consecutive series of 35 patients with DILS diagnosed from 1992 to 1995 in a cohort of 4100 outpatients infected with HIV-1. METHODS: Thirty-five individuals with DILS were ascertained from this cohort and followed for 720 patient-months. Clinical, serological and immunogenetic features of these patients were studied and their demographics were compared with the rest of the outpatient population. RESULTS: DILS was found to be more prevalent in African Americans (60%) than in Caucasians (26%) or Mexican Americans (14%) [odds ratio (OR), 2.32; 95% confidence interval (CI), 1-12-4.81; P = 0.02] and in persons with male-to-male transmission of HIV-1 (71%) (OR, 2.82; 95% CI, 1.29-6.29; P = 0.007). All patients had bilateral parotid gland enlargement. The majority had sicca symptoms. The most common extraglandular sites of disease were lung (31%), muscle (26%), and liver (23%). Four patients had biopsy-proven polymyositis. Thirteen patients met the 1993 Centers for Disease Control and Prevention case definition of AIDS. Sixteen (52%) patients expressed human leukocyte antigen (HLA)-DRS (DRB1*1102), DR6 (DRB1*1301, *1302), or DR7, and 11 (36%) expressed HLA-DR2. CONCLUSIONS: DILS is more common in African Americans and in persons with male-to male transmission of HIV-1. HIV-associated polymyositis appears to occur in the setting of DILS. PMID- 8728043 TI - Complications of varicella zoster virus reactivation in HIV-infected homosexual men. AB - OBJECT: To study the complication rate of varicella zoster virus (VZV) reactivation and the relationship between complications, presentation and localization of zoster and immune function in HIV disease. DESIGN AND METHODS: A total of 142 episodes of VZV reactivation in 113 out of 544 HIV-1-infected participants in the Amsterdam Cohort Study of homosexual men were studied. Persistent hyperkeratotic or necrotic skin lesions, post-herpetic neuralgia, other neurological events, ocular events and pneumonitis occurring within 6 months of the onset of the last episode of VZV reactivation were defined as complications, provided that other possible diagnoses were excluded and the event had been previously described in the literature as related to VZV reactivation. RESULTS: Twenty-four complications occurred in 15 (11%) of these 142 episodes. Complications occurred exclusively in the 40 episodes with either multidermatomal or disseminated presentation, or a trigeminal localization, or both. In the group of episodes of unidermatomal zoster at a non-trigeminal localization no complications occurred. Twenty-one episodes of herpes zoster were localized in the trigeminal area. Localization was not significantly associated with the level of immune function. Compared to unidermatomal presentation (n = 120), multidermatomal (n = 15) and disseminated presentation (n = 7) occurred at lower median CD4+ cell counts (330, 240 and 50 x 10(6)/l, respectively; P = 0.003) and significantly lower levels of CD3 monoclonal antibodies or phytohaemagglutinin induced T-cell reactivity in vitro. Complications were related to CD4+ cell counts, but in the cases of disseminated, multidermatomal or trigeminal zoster a CD4+ cell measurement provided no additional information on the risk of complications. CONCLUSION: In HIV-infected individuals the extent of the clinical presentation and the occurrence of complications of VZV reactivation are related to the degree of immunodeficiency. In episodes of VZV reactivation with either multidermatomal or disseminated presentation or a trigeminal localization, or both the complication rate was high. CD4+ cell counts provided no additional information on the complication risk. Oral acyclovir appears to be sufficient as therapy for unidermatomal zoster at a non-trigeminal localization. PMID- 8728044 TI - Trends in HIV infection among sexually transmitted disease patients in Paris. AB - OBJECTIVE: To assess trends in HIV infection among sexually transmitted disease (STD) patients. DESIGN: Repeated unlinked anonymous survey, 1991-1993. SETTING: STD clinics in Paris, France. SUBJECTS: Patients (n = 4354) with a new suspected STD. METHODS: HIV antibody testing, using blood from syphilis samples. RESULTS: HIV prevalences were stable over time in all transmission groups. One-third of homo-/bisexual men were HIV-positive. Prevalence was 2.5 times higher among heterosexual patients from Africa or the Caribbean than among those from other countries. Among patients under 25 years of age prevalence significantly decreased from 4.3% in 1991 to 0.8% in 1993 (P = 0.01). Among homo-/bisexual men, despite a 50% reduction in the incidence of STD, the absolute number of those newly HIV-infected remained stable; median age increased from 28 years in 1991 to 32 years in 1993 (P = 0.02). Among heterosexuals, trends in HIV incidence were difficult to assess: recently infected patients were more likely to be identified in 1993 than in 1991, since the proportion of patients who reported a recent HIV negative test increased over time. CONCLUSION: Prevalence studies contribute to define specific subgroups which should be targeted for prevention (HIV-positive or older homosexuals, heterosexuals from Africa and the Caribbean). Despite a decrease in both overall STD incidence and HIV prevalence among patients aged under 25 years, overall HIV incidence has not decreased, at least among homo /bisexual men in whom recent HIV infections occurred at a high rate overall, and increased in those aged 35 years or more. Sentinel site-based HIV seroprevalence studies are best interpreted in the light of results obtained from different populations and through routine surveillance of STD. PMID- 8728045 TI - AIDS: a major health problem among young Italian women. AB - OBJECTIVE: To estimate the extent of the HIV/AIDS epidemic among Italian women and to assess its impact on mortality. Further, to assess the incidence of AIDS and the main characteristics of women affected by AIDS: with particular attention to those aged 25-34 years, and to identify differences in these characteristics compared with men infected with AIDS. METHODS: Incidence data were derived from the Italian National Register of AIDS Cases (RAIDS). The most recent complete data refer to cases diagnosed to the end of 1994. Mortality data were collected from the Italian mortality database, which is held by the Italian National Census Bureau (ISTAT) and processed by the Laboratory of Epidemiology and Biostatistics at the National Institute of Health of Italy (Istituto Superiore di Sanita; ISS). The most recent mortality data refer to 1992, but projections were made to obtain mortality data up to 1994. All the rates were standardized by age and sex. RESULTS: A total of 26 957 cases of AIDS, 21 441 men (79.5%) and 5516 women (20.5%), diagnosed before the end of 1994 were reported to RAIDS. The majority of cases and deaths occurred in individuals aged 25-34 years. The incidence of AIDS among women increased from 2.1 per 100 000 in 1987 to 17.2 per 100 000 in 1994. The sex ratio (men:women) fell from 5.2 in 1987 to 2.9 in 1994. In 1992, AIDS was the second most common cause of death (slightly behind cancer and ahead of traffic accidents) in Italy, but was the most common cause in three regions of northern Italy (Liguria, Lombardia and Emilia). CONCLUSIONS: Although the AIDS epidemic in the past has predominantly affected young men, AIDS is now rapidly becoming a major health problem for young women in Italy. Moreover, as these women are at reproductive age, this may have important consequences in terms of increased morbidity and mortality among children. PMID- 8728046 TI - HIV-1, HIV-2, human papillomavirus infection and cervical neoplasia in high-risk African women. AB - OBJECTIVE: To determine the effect of HIV-1 and HIV-2 infection on the prevalence of cervical human papillomavirus (HPV) and squamous intraepithelial lesions (SIL) in a population of high-risk women in Senegal. DESIGN AND PARTICIPANTS: Cross sectional study among 759 female commercial sex workers, including 68 with HIV-1, 58 with HIV-2, 14 with HIV-1 and 2, and 619 without HIV infection. RESULTS: Overall, HPV was detected in 43% of women by polymerase chain reaction (PCR), and in 7% by Southern transfer hybridization, with 7.4% of all women having SIL. The mean CD4 count was 820, 1205, and 727 x 10(6)/l for those with HIV-1, HIV-2, and dual HIV-1 and 2 infections, respectively, and 1447 x 10(6)/l for those without HIV infection. Both HIV-1 and HIV-2 were associated with HPV, as detected by PCR [HIV-1 odds ratio (OR), 2.9; 95% confidence interval (Cl), 1.7-4.9; HIV-2 OR, 1.7; 95% Cl, 1.0-2.9]. HIV-2 was also associated with cervical SIL, and although the association between HIV-1 and SIL did not attain statistical significance, a trend was apparent (HIV-1 OR, 1.8; 95% Cl, 0.7-4.7; HIV-2 OR, 2.9; 95% Cl, 1.2 7.2). CONCLUSIONS: Despite less immunosuppression with HIV-2, both HIV-1 and HIV 2 were associated with detection of HPV. HIV-2 was also associated with SIL. Further studies are needed to examine the risks of high-grade SIL and invasive cervical cancer with HIV-1 versus HIV-2 infection. PMID- 8728047 TI - Progression of HIV: follow-up of Edinburgh injecting drug users with narrow seroconversion intervals in 1983-1985. AB - OBJECTIVE: To describe progression and survival of individuals infected with HIV by injecting drug use in Edinburgh. DESIGN AND METHODS: From 313 HIV-infected patients with retrospectively estimated narrow seroconversion intervals, 260 infected via injecting drug use in the years 1983-1985 were selected for the study group. MAIN OUTCOME MEASURES: The effects of gender, age, human leukocyte antigen (HLA) type and zidovudine (ZDV) treatment on progression and survival from seroconversion; Weibull estimates of the AIDS incubation distribution and the overall survival distribution; slopes of absolute CD4 lymphocyte loss (on the square root scale) and loss of CD4 percentage. RESULTS: The cumulative progression rates at 10 years were 68% to CDC stage IV and 31% to AIDS with a mortality rate of 25%. Three-year survival rates for AIDS and CDC stage IV cases were 25 and 72%, respectively. Gender and age effects on progression or overall survival were not found, although those aged over 30 years experienced poorer survival from AIDS. A strong HLA (A1, B8, DR3) association with faster progression and poorer survival was found. Median survival was estimated by Weibull distribution to be 12.6 years; median AIDS-free time was estimated to be 11.6 years. CD4 cell loss was approximately linear when transformed to the square root scale as was the decline in CD4 percentage. Only HLA effects on slopes were found: A1,B8, DR3 was significantly associated with faster loss of both absolute CD4 cells and CD4 percentage (P < 0.001) and B27 was significantly associated with slower loss of CD4 percentage (P = 0.01). CONCLUSIONS: Edinburgh IDU do not seem to progress more rapidly than other cohorts with predominantly different risk activities. Older age was associated with poorer survival from AIDS but no gender effect was found for progression or overall survival. The clearest significant association with AIDS progression, mortality and loss of CD4 cells was the phenotype HLA A1,B8,DR3. In contrast HLA B27 was associated with slower loss of CD4 cells. PMID- 8728049 TI - Similarity between Nef of primate lentiviruses and p15E of murine and feline leukaemia viruses. PMID- 8728048 TI - The Edinburgh cohort of HIV-positive injecting drug users at 10 years after infection: a case-control study of the evolution of dementia. AB - OBJECTIVE: To examine the evolution of dementia in HIV-positive injecting drug users (IDU) in Edinburgh. DESIGN: Case-control study. PARTICIPANTS: Twenty six (6%) out of 404 patients in the Edinburgh cohort of HIV-positive IDU who have developed HIV-1-associated dementia in the 10 years since infection and seroconversion. METHODS: Patients were tested repeatedly, where possible, on a range of neuropsychological and neurophysiological measures. The results from patients with dementia were compared with those of age, sex and IQ-matched non demented HIV-positive controls from the cohort. An auditory event-related potential (P3 or P300), a neurophysiological measure of cognitive function, detected the onset of a marked slowing of cognitive and psychomotor functions. Neuropsychological measures that involve the speed of information processing such as the Trail-Making task also identified the early stages of dementia. RESULTS: Dementia was associated with a more advanced stage of systemic disease, increased rates of decline in CD4 cell counts and markedly reduced survival compared with the non-demented controls. No evidence for a protective effect of treatment with zidovudine was detected. CONCLUSION: In the first 10 years after infection with HIV-1 dementia is an individual development, not the clinical extreme of general intellectual impairment, and had occurred in at least 6% of our IDU cohort. Future questions concern the long-term rate of dementia, the critical neuropathological change and the true potential for early treatment. PMID- 8728050 TI - Detection of Isospora in the stools of AIDS patients using a new rapid autofluorescence technique. PMID- 8728052 TI - The occurrence of Blastocystis hominis in HIV-infected patients. PMID- 8728051 TI - Testing and management of Toxoplasma serology in HIV-seropositive patients. PMID- 8728053 TI - Condom use in clients of prostitutes in Spain. PMID- 8728054 TI - Lack of utility of routine chest radiographs in HIV-infected patients. PMID- 8728055 TI - Antiplatelet antibodies during the course of HIV-preventive vaccine trial. PMID- 8728056 TI - CD4 count in HIV infection is positively correlated to interferon-gamma and negatively correlated to interleukin-10 in vitro production. PMID- 8728057 TI - Serum nitrite concentration suggests a role for nitric oxide in AIDS. PMID- 8728058 TI - Interest and reliability of a polymerase chain reaction on bone-marrow samples in the diagnosis of visceral leishmaniasis in AIDS. PMID- 8728059 TI - Isoscutellarein 7-glucosyl(1-->2)xyloside from sixteen species of Sideritis. AB - A new flavone diglycoside isolated from the aerial parts of Sideritis lutoela was characterized as isoscutellarein 7-O-beta-D-glucopyranosyl(1-->2) beta-D xylopyranoside by means of spectral analyses, applying 2D NMR techniques and NOE experiments. Its presence in 15 other species of Sideritis is described. PMID- 8728060 TI - Annosenegalin and annogalene: two cytotoxic mono-tetrahydrofuran acetogenins from Annona senegalensis and Annona cherimolia. AB - Two new cytotoxic mono-tetrahydrofuran acetongenins, annosenegalin and annogalene, have been isolated from the cytotoxic methanolic extract of Annona senegalensis and A. cherimolia seeds. Their structures were established on the basis of 1D and 2D NMR spectroscopic techniques. Annosenegalin belongs to the rare type of C37 mono-tetrahydrofuran acetogenins, and annogalene is an olefinic acetogenin. PMID- 8728061 TI - Flavonoids, cinnamic acids and coumarins from the different tissues and medicinal preparations of Taraxacum officinale. AB - Three flavonoid glycosides: luteolin 7-glucoside and two luteolin 7-diglucosides were isolated from dandelion flowers and leaves together with free luteolin and chrysoeriol in the flower tissue. The hydroxycinnamic acids, chicoric acid, monocaffeyltartaric acid and chlorogenic acid were found throughout the plant and the coumarins, cichoriin and aesculin were identified in the leaf extracts. This represents the first report of free chrysoeriol (luteolin 3'-methyl ether) in Taraxacum officinale agg. An earlier provisional identification of chicoric acid, chlorogenic acid, cichoriin and aesculin in a phenolic survey of the tribe Cichorieae is confirmed. Chicoric acid and the related monocaffeyltartaric acid were found to be the major phenolic constituents in flowers, roots, leaves and involucral bracts and also in the medicinal preparations tested. PMID- 8728062 TI - Podacycline A and B, two cyclic peptides in the latex of Jatropha podagrica. AB - Two novel cyclic peptides were isolated from the latex of Jatropha podagrica, which we named podacycline A and B. Podacycline A is a cyclic nonapeptide with the sequence Gly1-Leu2-Leu3-Gly4-Ala5-Val6-Trp7-Ala8-Gly9+ ++-Gly1. The sequence of podacycline B, a cyclic heptapeptide, was determined to be Phe1-Ala2-Gly3-Thr4 Ile5-Phe6-Gly7-Phe1. The amino acid residues of both compounds were found to have the L-configuration. PMID- 8728063 TI - An abietane diterpene and two phenolics from Salvia forskahlei. AB - From the roots of Salvia forskahlei a new diterpenoid, forskalinone, two new aromatic compounds, the octanol esters of cis- and trans-4-O-methyl-caffeic acid dimers, were isolated together with the known compounds stigmast-3-one, sitosterol and alpha-amyrin. The structures of the new and the known compounds were established by spectral data. The antimicrobial activity of forskalinone and the dimeric cinnamic acid esters was tested against standard bacterial strains and a yeast, namely Bacillus subtilis ATCC 6633, Staphylococcus aureus 6538P, S. epidermidis ATCC 12228, Proteus mirabilis ATCC 14153, Escherichia coli ATCC 8739, Klebsiella pneumonia ATC 4352, Pseudomonus aeruginosa ATCC 9027, Enterococcus faecalis ATCC 29212, beta-haemolytic Streptococcus and Candida albicans ATCC 10231. Forskalinone showed moderate resistance against S. epidermidis (670 micrograms ml-1) and slight activity against E. faecalis (168 micrograms ml-1). trans-4-O-Methyl-caffeic acid dimer octanol ester was inactive while the cis isomer showed a slight activity against C. albicans (156 micrograms ml-1). PMID- 8728064 TI - Furanoid diterpene glucosides from Tinospora rumphii. AB - The stems of Tinospora rumphii yielded clerodane type furanoid diterpene glucosides: the new rumphioside I, the known borapetosides C and F and three other compounds. Their structures were established on the basis of various spectroscopic techniques. The three compounds have physical and spectral data identical to those of the reported compounds borapetosides E, D and B, respectively. However, the positions of the lactone rings of the first two compounds and the configuration of the bond attaching the glucose moiety to C-6 of the third differ from the reported ones. PMID- 8728065 TI - Aryl and triterpenic glycosides from Margyricarpus setosus. AB - Investigation of aerial parts of Margyricarpus setosus afforded three new aryl glycosides, beta-hydroxyphenylethyl-O-alpha-L-rhamnopyranosyl (1-->6)-beta-D glucopyranoside, benzyl-O-alpha-L-rhamnopyranosyl (1-->6)-beta-D-glucopyranoside, 4-methoxybenzyl-O-alpha-L-rhamnopyranosyl (1-->6)-beta-D-glucopyranoside, and three new triterpene glycosides, tormentic acid 3 beta-O-beta-D quinovopyranoside, tormentic acid 3 beta-O-beta-D-fucopyranoside and tormentic acid 3 beta-O-alpha-L-rhamnopyranoside. Twelve known compounds were also found, among them beta-hydroxy-3',4'-dimethoxyphenylethyl glucoside and beta-hydroxy 3',4'-dimethoxyphenylethyl rutinoside, which were obtained previously by partial synthesis, were isolated for the first time from a natural source. The structures of the new compounds were elucidated on the basis of chemical and spectral data. PMID- 8728066 TI - Triacylated anthocyanins from Ajuga reptans flowers and cell cultures. AB - Four anthocyanins were isolated from Ajuga reptans flowers and one from the cell cultures. By FAB mass spectrometry measurements, the structures of these pigments were determined as delphinidin and cyanidin glucosides acylated with two cinnamic acids, while three of them were also malonylated. A delphinidin-based pigment in the crude extract from cell cultures was identical to the major flower pigment as shown by HPLC co-chromatography. Moreover, by application of 1H and 13C NMR consisting of DQF-COSY, NOESY, ROESY, 2D-HOHAHA, HSQC and HMBC methods, the structures of two new anthocyanins were identified as delphinidin and cyanidin 3 O-(2-O-(6-O-(E)-p-coumaryl-beta-D-glucopyranosyl)-(6-O-(E)-p- coumaryl)-beta-D glucopyranosyl)-5-O-(6-O-malonyl-beta-D-glucopyranoside ). The deacylated anthocyanins were confirmed as delphinidin and cyanidin 3-sophoroside-5 glucosides. PMID- 8728067 TI - Anhydroicaritin 3-O-rhamnosyl(1-->2)rhamnoside from epimedium koreanum and a reappraisal of other rhamnosyl(1-->2, 1-->3 and 1-->4)rhamnoside structures. AB - An anhydroicaritin 3-O-rhamnosylrhamnoside from Epimedium koreanum is defined as the 3-O-alpha-L-[alpha-L-rhamnopyranosyl(1-->2)rhamnopyranoside] on the basis of 2D NMR evidence. Complete assignments of the 1H and 13C NMR spectra of this compound are presented for the first time. The NMR distinction of 1-->2, 1-->3 and 1-->4 linked rhamnopyranosylrhamnopyranosides are discussed and indicate that baohuosides III and V from E. davidii and baohuoside VI from E. davidii and E. pubescens, respectively, are (1-->2) linked. PMID- 8728068 TI - An iridoid glucoside from dipsacus asperoides. AB - A new iridoid glucoside, loganic acid-6'-O-beta-D-glucoside, has been isolated from the defatted root of Dipsacus asperoides. Its structure has been elucidated by spectroscopic means as 1S-(1 alpha,4a alpha,6 alpha,7 alpha,7a alpha)-1-[(6-O beta-D- glucopyranosyl-beta-D-glucopyranosyl)oxyl]1,4a,5,6,7,7a-hexahydro-6-hydr oxy-7- methyl-cyclopenta[c]pyran-4-carboxylic acid. PMID- 8728070 TI - Efficacy of nonsurgical therapy for urinary incontinence. AB - OBJECTIVE: to assess the efficacy of aggressive nonsurgical therapy for urinary incontinence in women and to determine if it is advantageous to perform invasive urodynamic testing prior to initiating treatment. STUDY DESIGN: We subjectively assessed 202 incontinent women three months to four years following nonsurgical therapy for urinary incontinence. Therapy in all patients included behavioral modification in the form of timed voiding and pelvic floor exercises. Depending on the suspected diagnosis and menopausal state, patients were also treated with various combinations of pharmacologic agents and estrogen replacement therapy. Based on initial evaluation, patient preference and referring physician preference, 100 women were started on therapy after a minimal evaluation, while 102 women were treated based on invasive urodynamic testing. A subjective assessment of treatment outcome was made via a telephone interview in which patients were asked standardized questions. RESULTS: One hundred nineteen cases (59%) were subjectively improved or cured with nonsurgical therapy. Performing urodynamic testing prior to initiating therapy did not increase the overall success rate: 69 of 100 patients (69%) treated on the basis of history, physical examination and a minimal office evaluation were improved or cured, while only 50 of 102 patients (49%) having electronic multichannel urodynamic testing were improved or cured. CONCLUSION: All women with urinary incontinence should be initially offered nonsurgical therapy since a large percentage will obtain satisfactory results. It is not beneficial or cost-effective to perform electronic urodynamic testing prior to initiating therapy. PMID- 8728071 TI - Laparoscopic uterine biopsy for diagnosing diffuse adenomyosis. AB - OBJECTIVE: To investigate the possibility of diagnosing diffuse adenomyosis with the uterus in situ. STUDY DESIGN: Myometrial needle biopsy was performed on a prospectively evaluated series of 72 women undergoing laparoscopy for infertility and/or chronic pelvic pain. Before surgery the patients completed a questionnaire on the presence and severity of dysmenorrhea according to a linear analog and verbal rating scale. A single specimen was taken from along the median line in the upper third of the posterior uterine wall using a 14-gauge Tru-cut needle inserted through the anterior abdominal wall under laparoscopic guidance. RESULTS: No complications occurred. Adenomyosis was diagnosed in 8 of the 42 subjects with menstrual pain and 5 of the 30 asymptomatic ones (19% versus 17%) and pelvic endometriosis in 27 and 10, respectively (64% versus 33%, P = .02). The frequency of moderate or severe dysmenorrhea was greater in women with adenomyosis than in those without adenomyosis or endometriosis, but the difference was not statistically significant. Conversely, menstrual pain was significantly more frequent and severe in patients with endometriosis as compared with women who had neither endometriosis nor adenomyosis. CONCLUSION: Uterine needle biopsy is simple, rapid and safe, but its overall clinical importance, as well as the predictive value of a normal histologic result, requires further investigation. PMID- 8728072 TI - Laparoscopy vs. laparotomy for gynecologic procedures. Impact on resident training. AB - OBJECTIVE: To compare laparoscopic approaches to traditional ones for operative procedures. STUDY DESIGN: Operating residents collected data on all abdominal, vaginal and laparoscopic vaginal hysterectomies, myomectomies, cystectomies/oophorectomies and ectopics between December 1, 1992, and July 31, 1993. Statistical analysis included Student's t test for interval data and chi (2)/Fisher's exact test for categorical data. RESULTS: Leiomyomas were the major indications for all three types of hysterectomy. Patients selected for vaginal hysterectomy had significantly higher parity and older age than those scheduled for abdominal hysterectomy or laparoscopically assisted vaginal hysterectomy (LAVH). Abdominal hysterectomy uteri were larger than those using vaginal or LAVH procedures. LAVH procedures were significantly longer when failed laparoscopic procedures were excluded. Abdominal hysterectomy patients used twice as much morphine as vaginal and LAVH patients, and the length of stay was longer. LAVH was significantly more expensive than vaginal hysterectomy. There was no difference in operating time for nonhysterectomy cases, but the cost and length of stay were less for laparoscopy. CONCLUSION: Laparoscopy can be advantageous for certain procedures but has little advantage for hysterectomy and may make fewer vaginal hysterectomies available for resident education since the patients selected for LAVH and total vaginal hysterectomy are more similar to each other than to patients selected for total abdominal hysterectomy. PMID- 8728073 TI - Laparoscopically assisted vaginal hysterectomy. Single-surgeon technique with minimal assistance. AB - OBJECTIVE: To prospectively evaluate parameters of surgical outcome of a novel technique of laparoscopically assisted vaginal hysterectomy performed by a single surgeon with minimal surgical help from an inexperienced assistant. STUDY DESIGN: Four women who had indications for hysterectomy but relative contraindications to the strictly vaginal approach were consecutively entered into the study. The hysterectomy was accomplished with the Endo GIA 30, O'Connor-O'Sullivan self retaining vaginal retractor, the ligature vaginal hysterectomy technique and one surgical assistant. RESULTS: The average operative time was 133 minutes (range, 105-167). Blood loss averaged 288 mL (range, 150-350). The average length of hospitalization was 62 hours (range, 36-72). The average time of convalescence was two weeks (range, one to three). CONCLUSION: Preliminary results from a series of four patients suggest that this procedure may be a viable and safe alternative for the single surgeon. PMID- 8728074 TI - Obstetric management of incompetent cervix and bulging fetal membranes. AB - OBJECTIVE: To evaluate the effect of emergency cerclage on perinatal outcome in patients with incompetent cervix. STUDY DESIGN: In this descriptive retrospective study, the charts of all patients admitted from 1985 to 1992 with a diagnosis of incompetent cervix and cervical dilatation > or = 2 cm were reviewed. Cervical dilatation at the time of cerclage placement, subsequent pregnancy duration and neonatal outcome were analyzed. Statistical analysis was performed using the paired Student t test. RESULTS: During the eight-year period, 42 patients were admitted with a diagnosis of incompetent cervix, cervical dilatation > or = 2 cm, and underwent emergency cerclage. Cerclage resulted in a median (range) continuation of pregnancy of 30 (2-102) days. In 20 of these patients the membranes were not bulging through the external cervical os, and pregnancy was extended a median of 50 (30-102) days. The remaining 22 patients presented with membranes bulging into the vagina, and emergency cervical cerclage resulted in a median pregnancy extension of 16 (2-98) days. Parturients with bulging membranes were then analyzed according to cervical dilatation. Cerclage success was significantly lower when attempted at a cervical dilation > or = 4 cm. Median pregnancy duration was 6 (2-15) versus 21 (5-98) days in patients with cervical dilatation < 4 cm. CONCLUSION: The results of this study support emergency cerclage in patients without bulging membranes. The data do not support the use of emergency cervical cerclage in patients with bulging membranes and dilatation > or = 4 cm. In those patients, the use of cerclage should be individualized. PMID- 8728075 TI - Cell surplus on sampling devices after routine cervical cytologic smears. A study of residual cell populations. AB - OBJECTIVE: To demonstrate whether considerable valuable cellular material remains on the sampling device after a conventional cervical cytologic smear and to investigate whether evidence of cervical disease may be present on the sampling device but missed because it is not represented on the first slide. STUDY DESIGN: Routine cervical cytologic smear material was taken from 60 women who were attending the colposcopy or gynecologic oncology clinic at New England Medical Center. At the time of the colposcopic examination, a cervical scrape was made using a Cytobrush followed by a Cervex brush. After the smear was made from each device, the sampling devices were also used to make four additional slides. Both the Cytobrush and Cervex brush were applied to each slide. RESULTS: Cytologically, all five slides were satisfactory for screening in the 60 cases. In all diagnostic cases but one, disease was well represented across all five slides. CONCLUSION: Cellular material collected in a cervical scrape is sufficient to prepare at least five satisfactory smears. The one case where an abnormality was detected only on the third slide suggests the possibility of nonrandom subsampling and of discarding the majority of the cellular material from the patient. PMID- 8728076 TI - The shapes of the female pelvis. Contributing factors. AB - OBJECTIVE: To determine the etiologic factors that play a role in shaping the female pelvis. STUDY DESIGN: The backgrounds of 611 pregnant women at term who underwent x-ray pelvimetry between 1962 and 1980 at term were extensively investigated. RESULTS: Several facts were statistically significant. First, 24.1% of the patients had an android pelvis, although only 5.7% presented any sign of hyperandrogenism. Signs of hyperandrogenism were similarly encountered in patients with a gynecoid and any other type of pelvis. Second, the android pelvis was encountered mostly in patients exposed to strenuous physical activity during adolescence. Third, the anthropoid pelvis was encountered more often when the acquisition of erect posture was delayed beyond the usual age of 14 months, while a platypelloid pelvis was more frequent when erect posture was acquired before 14 months. CONCLUSION: The final shape of the female pelvis seems to be determined by culture and environment as well as by genetics. PMID- 8728077 TI - Endometrial rollerball ablation. AB - OBJECTIVE: To study a standardized technique for endometrial rollerball ablation and various methods of preoperative preparation. STUDY DESIGN: All patients had refractory symptomatic menorrhagia and previously failed conservative surgery and other forms of medical therapy, such as progestogens. Patients received a preoperative regimen of either leuprolide acetate, danazol, Nolvadex or Depo Provera. They then underwent hysteroscopically controlled rollerball ablation at 80-100 W with 1.5% glycine as the distending medium. The endometrium was evaluated hysteroscopically and considered to be completely atrophic, intermediate or no response. RESULTS: Refractory symptomatic menorrhagia was treated successfully in 38 of 40 patients. Nineteen reported amenorrhea; the other 21 reported subjective and objective improvement of bleeding. Three patients, despite improvements in flow, were unhappy with the overall result. Two did not wish repeat ablation and subsequently underwent vaginal hysterectomy. The third underwent repeat ablation and became amenorrheic. The ability to achieve complete endometrial atrophy prior to ablation was improved with leuprolide acetate (19/24) and danazol (5/6) when compared to tamoxifen (0/4) and Depo Provera (0/6). Attainment of amenorrhea after ablation was significantly improved when complete atrophy (19/24) was achieved prior to ablation as compared to the ability to achieve amenorrhea when no endometrial response was achieved (0/7). The only significant complication was one uterine perforation in a patient undergoing repeat ablation. CONCLUSION: Endometrial rollerball ablation is a safe, effective means of controlling refractory menorrhagia. Amenorrhea is best attained when complete preoperative atrophy is achieved. Leuprolide and danazol were superior to tamoxifen and Depo-Provera. PMID- 8728078 TI - Primary hyperaldosteronism in pregnancy. A case report. AB - BACKGROUND: Primary hyperaldosteronism is an uncommon disorder, and there are few reports of its occurrence and management in pregnancy. CASE: Primary hyperaldosteronism was suspected before pregnancy in a 31-year-old women with refractory hypertension and hypokalemia. Pregnancy evaluation revealed suppressed renin levels and high aldosterone levels; computed tomography revealed a 1-cm mass in the left adrenal gland. The patient became pregnant before completion of evaluation and treatment. On high doses of nifedipine and nadolol, the first trimester blood pressure was 130/98 mm Hg and remained high in the early second trimester. In view of the risks of poorly controlled hypertension, adrenalectomy was performed at 15 weeks' gestation, with rapid improvement in blood pressure and elimination of the patient's requirement for large doses of potassium daily. Antihypertensive medication was withdrawn, with maintenance of normal blood pressure until 36 weeks' gestation. At that time the blood pressure rose slightly but responded to bed rest. A healthy female infant was delivered at term by cesarean section. CONCLUSION: Previous reports of emergency preterm delivery and a case of neonatal mortality in the setting of hyperaldosteronism in pregnancy confirm the significant risks associated with this condition. In our patient, adrenalectomy in the early second trimester resulted in a rapid and sustained improvement in hypertension, reversal of hypokalemia and a good pregnancy outcome. PMID- 8728079 TI - Use of a gonadotropin-releasing hormone agonist in the evaluation of postmenopausal virilization due to ovarian hyperthecosis. A case report. AB - BACKGROUND: Hyperthecosis in a postmenopausal woman is a very rare cause of virilization, and only five cases have been reported previously. CASE: A woman presented with a nine-year history of increasing hirsutism and a mild virilization beginning in the perimenopausal period. Initial androgen metabolite concentrations suggested attenuated late-onset adrenal hyperplasia, but a trial of dexamethasone treatment was ineffective. Subsequent use of leuprolide acetate resulted in a biochemical and clinical improvement in the signs and symptoms. CONCLUSION: This case is unique because gonadotropin-releasing hormone agonist administration was utilized as both a diagnostic and therapeutic modality. PMID- 8728080 TI - Clinical and DNA studies on 46, XY females with gonadal dysgenesis. A report of six cases. AB - BACKGROUND: Understanding the process of sex determination has been aided by the molecular analysis of individuals whose karyotype does not correspond to their phenotype, 46, XX males and 46, XY females. CASES: We studied the clinical and molecular data on six 46, XY females of Indian ethnic origin. In each subject, cytogenetic analysis indicated a 46, XY karyotype without mosaicism. In four of the cases DNA studies were performed on the sex-determining region, Y chromosome gene. A de novo point mutation was identified in one subject. CONCLUSION: Our data provide additional evidence for genetic heterogeneity in the etiology of 46, XY gonadal dysgenesis. PMID- 8728081 TI - Dermatofibrosarcoma protuberans of the vulva. A case report. AB - BACKGROUND: Dermatofibrosarcoma protuberans (DFSP) is a cutaneous malignancy that is uncommon anywhere on the body but is especially uncommon on the vulva. This cancer rarely metastasizes but has a high incidence of local recurrence because of its tendency to grow in fingerlike projections away from the primary tumor. CASE: A 36-year-old, black woman was the 10th and youngest patient with DFSP of the vulva to be reported. Treatment of the 5-cm, mobile, well-circumscribed lesion consisted of local excision. Several adjacent microscopic lesions showing DFSP were found in the pathologic specimen, however, so an even wider excision was performed. Twenty-seven months following removal, there was no evidence of recurrence. CONCLUSION: Survival rates for DFSP of the skin range from 91% to 100% in reported series, but local recurrence rates of 20-49% have also been noted. The reported deaths from the disease have resulted from extensive local spread due to inadequate excision and, only rarely, metastases. Because DFSP tends to spread to microscopic projections away from the visible lesions, very wide local excision is required for tumor control. PMID- 8728082 TI - Bilateral laparoscopic gonadectomy in a patient with complete androgen insensitivity. A case report. AB - BACKGROUND: Complete androgen insensitivity (CAI) is the most common cause of male pseudohermaphroditism. Affected individuals are advised to have bilateral gonadectomies to prevent the development of germ cell tumors. CASE: A 19-year-old phenotypic female presented for the evaluation of primary amenorrhea and was diagnosed with CAI. Operative videolaparoscopy was performed, and the intraabdominal gonads were resected; the patient was discharged in < 24 hours. CONCLUSION: Laparoscopic gonadectomy can be performed in individuals with CAI. This approach eliminates the need for laparotomy and results in rapid recovery with minimal blood loss. This approach should be considered for all patients with CAI and intraabdominal gonads. PMID- 8728083 TI - Pelvic tuberculosis presenting as an asymptomatic pelvic mass with rising serum CA-125 levels. A case report. AB - INTRODUCTION: The incidence of tuberculosis is rapidly rising. This disease may present in an atypical manner, and physicians must reacquaint themselves with its protean manifestations. CASE REPORT: A 41-year-old woman presented with an asymptomatic pelvic mass and rising levels of serum CA-125. At operation she had bilateral adnexal masses and generalized military peritoneal lesions, initially mistaken for ovarian carcinoma. The final diagnosis demonstrated tuberculosis. CONCLUSION: Tuberculosis must enter into the diagnosis of a pelvic mass. PMID- 8728084 TI - Retroperitoneal abscess after normal delivery. A report of two cases. AB - BACKGROUND: Retroperitoneal abscess is a rare complication of normal labor and delivery, and no apparent etiology is evident. CASES: We report on two patient who presented with back and leg pain soon after normal deliveries. While the condition was initially clinically misdiagnosed, ultrasonography and computed tomography demonstrated retroperitoneal collections and aided in guided percutaneous drainage. After a protracted course of antibiotic treatment and daily irrigation, the collections resolved. CONCLUSION: Ultrasonography and computed tomography were invaluable in the diagnosis and guided drainage of rare psoas abscesses complicating delivery. PMID- 8728085 TI - Uterine prolapse after laparoscopic uterosacral transection in nulliparous airborne trainees. A report of three cases. AB - BACKGROUND: Laparoscopic uterine nerve ablation (LUNA) has become a common alternative therapy for refractory dysmenorrhea. Few long-term complications have been reported. CASES: Severe uterine prolapse was diagnosed in three young female soldiers during or after the rigors of airborne training. All three had previously undergone LUNA procedures. No other risk factors for uterine prolapse could be identified in these cases. CONCLUSION: The etiology of uterine prolapse is complex, and although no conclusions as to cause and effect can be made, these cases suggest that LUNA should be performed with caution on women whose occupation or life-style is associated with heavy physical labor or exercises producing marked increases in intraabdominal pressure. PMID- 8728086 TI - Basal cell carcinoma of the vulva. A report of four cases. AB - BACKGROUND: Basal cell carcinoma is the most common malignant tumor of the skin, with approximately 400,000 new cases yearly in the United States. Basal cell carcinomas account for 2-3% of all vulvar malignancies. CASES: Four patients presented in the eighth and ninth decades of life (70, 78, 87 and 89 years). Seventy-five percent of patients had a unifocal lesion. Cases 3 or 4 presented with recurrent lesions at 5 and 10 years, respectively. All patients were treated with wide local excision. Surgical margins were free of disease. None of the patients had lymph nodes suspicious for malignancy. CONCLUSION: Basal cell carcinoma can present as a unifocal or multifocal lesion. The lesions are usually located on the labia majora. Patients are frequently diagnosed with basal cell carcinoma of the vulva in the eighth and ninth decades of life. Treatment consists of wide local excision. Although 50% of these cases recurred, the lesions were reexcised, with wide local resection. No metastatic lesions were identified in any of the patients. PMID- 8728087 TI - Pregnancy after the fontan procedure for tricuspid atresia. A case report. AB - BACKGROUND: Pregnancy is contraindicated in uncorrected tricuspid atresia. Even following palliative surgery with the Fontan procedure, only four pregnancies have been reported. CASE: A 32-year-old female presented at 8 weeks' gestation in her first pregnancy with a history of the Fontan procedure performed seven years earlier for cardiac decompensation despite two previous palliative procedures for tricuspid atresia. She remained class I with no deterioration and required no medication throughout the pregnancy. Amniocentesis for positive maternal alpha fetoprotein screening was normal. The pregnancy was complicated by antepartum hemorrhage at 29 weeks and premature rupture of membranes at 33 weeks, resulting in preterm labor and delivery of a 2.5-kg infant five days later. Apart from postpartum hemorrhage, the puerperium was uneventful, and the patient was well 20 weeks after delivery. CONCLUSION: Five similar pregnancies, including this one, have been reported. Although all were successful, three infants were preterm and two growth retarded. This observation suggests that for these patients, pregnancy must be monitored closely even though the mothers may be hemodynamically stable. PMID- 8728088 TI - Colovaginal fistula. Report of a case with failure of transvaginal repair. AB - BACKGROUND: Spontaneous development of colovaginal fistula is a rare cause of abnormal vaginal discharge in older women and most commonly arises from diverticular disease. Laparotomy for segmental colonic resection is the accepted treatment. The author found no previous reports of attempted transvaginal closure of these fistulae, although such closure of similar fistulae can sometimes be accomplished in patients with Crohn's disease when quiescence is achieved. CASE: An otherwise healthy 69-year-old female developed a colovaginal fistula as a result of diverticular disease. Resolution of all associated inflammatory changes was achieved over four months. A three-layer transvaginal closure of the fistula using the Futh technique was then performed; it was technically successful but broke down one week later. CONCLUSION: Transvaginal closure of a colovaginal fistula, even with intensive preparatory efforts, cannot be recommended on the basis of this attempt. PMID- 8728089 TI - Intraabdominal bleeding following laparoscopic adnexal surgery. A report of three cases. AB - BACKGROUND: The incidence of major complications from laparoscopy is reported to be 0.6-2.5%. The literature reviewing laparoscopic adnexal surgery reports few cases of intraoperative or postoperative complications. CASES: We describe three cases of intraabdominal bleeding following laparoscopic oophorectomy and salpingo oophorectomy and salpingo-oophorectomy. In two of the three cases, postoperative exploratory laparotomy failed to identify the source of bleeding. CONCLUSION: Possible causes of postoperative bleeding are loosening of clips from an automated stapling device and delayed bleeding from vessels tamponaded by the pneumoperitoneum. Many complications of operative laparoscopy probably are underreported and, when present, may occur outside a health care facility since many of these cases are performed on an ambulatory basis. Some patients will acquire medical and/or surgical intervention. PMID- 8728090 TI - Classification of hourglass-shaped fetal membranes. PMID- 8728091 TI - Puerperal inversion of one uterine horn. PMID- 8728092 TI - Formation of singlet oxygen in the thermal decomposition of 3-hydroxymethyl-3,4,4 trimethyl-1,2-dioxetane, a chemical source of triplet-excited ketones. AB - Triplet-excited carbonyl species can be generated by photoexcitation of carbonyl compounds or alternatively by thermal decomposition of 1,2-dioxetanes. Such electronically excited species are involved in the oxidative modification of biologically important molecules such as DNA. This study demonstrates that thermal decomposition of 3-hydroxymethyl-3,4,4-trimethyl-1,2-dioxetane (HTMD), which is a chemical source of triplet-excited ketones, is accompanied by infrared photoemission at 1270 nm characteristic for singlet oxygen monomol emission. The intensity of infrared photoemission at 1270 nm was solvent-dependent, in agreement with the reported lifetime of singlet oxygen in the employed solvents. Calibration of monomol emission with the endoperoxide of 1,4-dimethylnaphthalene (DMNO2) showed that HTMD (30 mM) produced 25 microMs of singlet oxygen. Thus, the yield of singlet oxygen production in the thermal decomposition of HTMD in carbon tetrachloride is about 0.1%. PMID- 8728093 TI - An antibody specific to N-terminal amino acid sequence of human maternal serum cystine aminopeptidase and its applications. AB - An antibody directed against a synthetic peptide corresponding to the N-terminal sequence of human cystine aminopeptidase (CAP) present in maternal serum was prepared. Although the antibody did not immunoprecipitate the activity of CAP, it was useful for purification and immunoblot analysis of CAP protein. An antipeptide antibody-conjugated Sepharose 4B column was very effective in purifying a single CAP protein from partially purified enzyme preparation, and Western blotting confirmed the binding of the antibody to CAP protein. PMID- 8728094 TI - Butyrate-induced alteration in lipid composition of human colon cell line HT-29. AB - The effect of butyrate-induced differentiation on lipid composition of HT-29 cells were studied. It was observed that cell differentiation was associated with increased activity of alkaline phosphatase and changes in the lipid composition. Differentiated cells showed increased level of triacylglycerol, cholesteryl esters and decreased free cholesterol and phospholipids. Changes were also observed in individual phospholipid composition. It appears that butyrate induced differentiation is associated with increased esterification of neutral lipids. PMID- 8728095 TI - A novel multigene family that the gene for a maize DNA-binding protein, MNB1a belongs to: isolation of genomic clones from this family and some aspects of its molecular evolution. AB - MNB1a is a DNA-binding protein of maize that contains a unique DNA-binding domain, designated the Dof domain, and its gene belongs to a multigene family. In putative proteins encoded by cDNA clones of the multigene family, the Dof domain was strongly conserved. In this study, two maize genomic clones of this multigene family were isolated. Although these clones did not contain the MNB1a gene, RNA transcripts corresponding to genomic clones were detected, suggesting that the clones encode authentic proteins. These clones contained sequences homologous to a portion of the sequence of the MNB1a cDNA. Unexpectedly, however, this region did not correspond to the Dof domain but to a region just downstream of the Dof domain. Although these observations seemed to suggest that the MNB1a gene family might have evolved via recombination between a DNA-binding domain and other functional domains, the MNB1a gene seemed to have no introns in the coding region. These results suggest that a simple exon-shuffling model is not sufficient to explain the molecular evolution of this multigene gene family but that complicated recombination played an important role in the evolution. PMID- 8728096 TI - Effects of aluminum(III) on catechol-stimulated nerve growth factor biosynthesis by cultured mouse brain astroglial cells. AB - The effects of aluminum and other metal ions on the synthesis of nerve growth factor (NGF) by cultured mouse brain astroglial cells have been investigated. Natural NGF formation was dependent on the AlCl3 concentration of the cell culture medium; it was stimulated at low concentrations but was inhibited at higher concentrations. Catechol-stimulated NGF formation was also inhibited at AlCl3/catechol molar ratios > 0.3, whereas ZnCl2 and CaCl2 had no effect under similar conditions. Ethylenediamine-N,N,N',N'-tetraacetate (EDTA) and citrate blocked the inhibitory effect of Al(III). These observations were explained by the complex formation between Al(III) and catechols. PMID- 8728097 TI - EPR-spin trapping kinetic studies of superoxide radicals produced by photosensitized hypocrellin A. A photodynamic therapeutic agent. AB - Kinetics of superoxide radicals generation by a photodynamic therapeutic agent, Hypocrellin A has been studied by EPR-spin trapping technique. The rate constant (k(g)) for superoxide radicals generation by Hypocrellin A was found to be 5.0 x 10(-3) s-1. The first-order rate constant (k1) and the half-life time (t0.5) for DMPO-O2- decay were 2.86 x 10(-3) s-1 and 242.3 s, respectively. It appears that oxyradicals, generated during the photodynamic process, may be associated with the treatment of various skin diseases by photodynamic therapy of Hypocrellin A. PMID- 8728098 TI - Mitochondrial DNA mutations associated with the 11778 mutation in Leber's disease. AB - To clarify the characteristics of possible synergestic mitochondrial DNA (mtDNA) mutations associated with Leber's hereditary optic neuropathy (LHON), we analyzed the entire nucleotide sequences of mitochondrial genome of two Japanese patients from independent pedigrees harboring the 11778 mtDNA mutation, and compared their sequences with those of 47 disease and 6 normal controls. We have detected several unique mutations in the mtDNA in addition to the 11778 mutation. Two nucleotide substitutions, an A-to-G transition at position 856 in the 12S rRNA gene and an A-to-G transition at 14692 in the T psi C loop of the tRNA(Glu) gene, occurred at highly conserved sites among various species. These mutations in combination with the 11778 mutation might synergetically contribute to the pathogenesis of LHON. PMID- 8728099 TI - Aggregation of phospholipid vesicles induced by the ribosome inactivating protein saporin. AB - Saporin-S6(SO-6) is a single chain ribosome inactivating protein, which can inhibit protein synthesis by inactivating eukaryotic ribosomes. The interaction of SO-6 with phospholipid model systems was described. SO-6 can specifically interact with negatively-charged phospholipid vesicles and it induces the aggregation of the lipid vesicles. The kinetics of the vesicle aggregation induced by SO-6 was studied. The saturating protein/lipid molar ratio was determined to be 1:100 based on titration experiments. The aggregation is dependent on the temperature in a range that was many times higher than the phase transition temperature of the phospholipid. The effect of pH on the aggregation of the vesicles can not be explained by simple deprotonation of side chain amino groups of the protein, and may be related to conformational changes of the protein. The maintenance of physiological ionic strength was required for the aggregation of SO-6 with vesicles. Finally, the interaction was prompted by Ca2+ ions, and was totally inhibited by EDTA, which suggests that SO-6 may interact with phospholipid vesicles in a Ca(2+)-dependent manner. PMID- 8728100 TI - Interaction of purified protein kinase C with key proteins of energy metabolism and cellular motility. AB - The phospholipid dependent protein kinase C is involved in regulation of cellular motility and energy metabolism. To study a possible direct interaction of protein kinase C with cellular motility and energy metabolism, we used purified rat brain protein kinase C to phosphorylate key proteins of these systems. Protein kinase C phosphorylates with comparable stoichiometry the G- but not the F- form of muscle and brain actin, the key protein of cellular motility. Glyceraldehyde-3-phosphate dehydrogenase, creatine kinase and glutamine synthase, key enzymes of energy metabolism, are also phosphorylated at comparable stoichiometry. The data suggest that protein kinase C might be directly involved in the regulation of cellular motility and energy metabolism. PMID- 8728101 TI - Effect of ceruloplasmin on 6-hydroxydopamine oxidation. AB - The effects of ceruloplasmin, a blu copper-containing serum glycoprotein, on the oxidation of the neurotoxin 6-hydroxydopamine in several chemical environments were studied. The spontaneous autoxidation of 6-hydroxydopamine proceeded by a free radical chain reaction involving O2-. and produced the corresponding chromogen 6-hydroxydopaminequinone and hydrogen peroxide. The process was accelerated in the presence of horse plasma ceruloplasmin. Yields of hydrogen peroxide in the presence of ceruloplasmin were significantly less than those measured in its absence. This is the first evidence of oxidase activity of ceruloplasmin toward a natural substrate, the 6-hydroxydopamine. PMID- 8728102 TI - Kinetic properties and the mechanism of activation of NAD-dependent glutamate dehydrogenase from Dictyostelium discoideum. AB - Two forms of the NAD-dependent glutamate dehydrogenase were partially purified from Dictyostelium discoideum, an activated and a non-activated form. V(max) for the non-activated enzyme was stimulated 88-fold and the activated enzyme 3-fold by 0.1 mM AMP (at their pH optima). Half maximal stimulation by AMP is achieved at 221 +/- 39 microM for the non-activated enzyme and 20 +/- 2 microM for the activated enzyme. We have shown that activation of NAD-GDH in vivo has many similarities to trypsin treatment of non-activated enzyme and that proteolysis is the probable mechanism of activation. PMID- 8728103 TI - Cyclo(-GLY-DSIP), A cyclic analog of the delta-sleep-inducing peptide: computer simulation of spatial structure involving NMR data. AB - The spatial structure of cyclo(-Gly-DSIP-), a physiologically active analog of the delta sleep-inducing peptide, was determined by computer modelling using 1H NMR data. An interesting feature of the spatial structure in DMSO was detected. One side of almost planar resulting conformation is formed by the side chains of the Asp5, Ser7 and Glu9 residues, the side chain of the Trp1 residue forming the other part of the outer surface. This feature may be associated with the functional properties of the peptide. PMID- 8728104 TI - Alzheimer's peptides A beta 1-40 and A beta 1-28 inhibit the plasma cholesterol esterification rate. AB - The amyloid fibrils of Alzheimer's disease and Down's syndrome amyloid deposits are composed mainly of aggregated amyloid beta protein (A beta) which also exists in a soluble form. It has been shown that both Alzheimer's disease and Down's syndrome share another common feature: the decrease in plasma cholesterol esterification in affected individuals. In the present work the effect of synthetic peptides A beta 1-40 and A beta 1-28 on normal human plasma cholesterol esterification rate was studied. Both peptides at a concentration of 1 ng/ml inhibited plasma cholesterol esterification rate to 40-50 % of control value. Statistical analysis showed no differences in the effect of A beta 1-40 and A beta 1-28 on the inhibition, suggesting the importance of A beta sequence 1-28 for this effect. PMID- 8728105 TI - A novel Bacillus intermedius extracellular alkaline phosphatase: isolation, physico-chemical and catalytic characteristics. AB - A new alkaline phosphatase was obtained as homogeneous preparation from culture filtrate of the spore-forming Bacillus intermedius. B. intermedius phosphatase was shown to be monomer with molecular weight of 47 kDa. The enzyme possesses phosphomonoesterase and phosphodiesterase activities and exhibits a broad specificity towards a wide variety of substrates. The purified phosphatase had an optimum temperature of 50 degrees C, optimum pH of 9.5 and was stable until 60 degrees C at pH 8-10. The effect of divalent metal ions and thiol reagents on catalytic activity of the enzyme was studied. PMID- 8728106 TI - Expression of mRNA encoding neurotrophic factors and its regulation in a hybrid neuronal cell line. AB - In a cultured hybrid neuronal cell line (BIM) which was produced between human neuroblastoma cells (IMR32) and thymidine auxotrophs (B3T) of rat nerve-like cells (B103), the mRNAs encoding ciliary neurotrophic factor (CNTF) and neurotrophins were detected by the polymerase chain reaction method. The conditioned medium of BIM cells enhanced choline acetyltransferase (ChAT) activity in septal neurons and survival of ciliary ganglion neurons. The mRNA expression of CNTF and neurotrophins in BIM cells was differently regulated by the stimulation with cAMP, FGF and retinoic acid. These data suggest multiple regulation and collaboration of neurotrophic factors. PMID- 8728107 TI - Structural changes of beta-lactoglobulin B induced by urea. Evidence of residual structure. AB - Several spectroscopic methods have been used to study the structure of beta lactoglobulin B at pH 2.1 in the presence of 8M urea. Fluorescence and polarization of fluorescence spectroscopy measurements indicate that the two tryptophanyl residues of the protein are exposed to the solvent in the denatured state. CD in the far-UV indicates that the amount of secondary structure in the denatured state is comparable to that found in the native state, whereas the CD spectrum in the near-UV shows that the tertiary structure is not completely disordered. The results of one-dimensional 1H NMR spectroscopy show that some local non-random structure is maintained in the denatured state, but most of the polypeptide chain has an extended non-globular conformation under the conditions of the present experiments. This conclusion is reinforced by the results of two dimensional 1H NMR conducted on denatured samples of beta-lactoglobulin B. The study of states with intermediate levels of order will aid the understanding of how the native structure of beta-lactoglobulin B is organised during the refolding pathways. PMID- 8728108 TI - 6-phosphogluconate dehydratase from Zymomonas mobilis: an iron-sulfur-manganese enzyme. AB - The enzyme 6-phosphogluconate dehydratase has been isolated in a stable form by a simple one-step procedure using dye ligand chromatography. The role of metal ions in the activity and stability of the enzyme was investigated. As with aconitase and several other dehydratase enzymes, the active site includes an Fe4S4 cluster. In addition, the purified enzyme has been shown to contain one manganese ion per subunit, which is also essential for activity. Rapid inactivation by superoxide radical was observed, which could only partly be protected by manganous ions The purified enzyme could be stabilised by alpha-glycerophosphate in place of manganese; glycerophosphate mimics the carbon atoms 4 to 6 of the natural substrate. This suggests that the manganous ion may involved in binding this part of the substrate. PMID- 8728109 TI - Cloning and sequence analysis of a putative transcription factor (MTF1) gene from Mucor circinelloides. AB - We describe here the fortuitous cloning of a putative transcription factor gene (MTF1) from the dimorphic fungus Mucor circinelloides. Sequence analysis of MTF1 revealed an open reading frame (ORF) of 1059 nucleotides encoding a protein of M(r) 39601. The deduced amino acid sequence from the ORF imparts two glutamine rich stretches which are homologous to a number of transcription factors characterized previously from various organisms. A Southern blot analysis of Mucor genomic DNA digested with different restriction endonucleases and probed with the 1.9 kb EcoR1 fragment of the putative transcription factor gene shows a single copy number of the the gene. Northern analysis during morphogenetic changes in Mucor suggested constitutive expression of the gene. PMID- 8728110 TI - High-level expression of functional human gamma-glutamyl transpeptidase using the baculovirus system. AB - The understanding of the structure and function of gamma-glutamyl transpeptidase (GGT) has been hindered by the difficulty of obtaining large quantities of functional enzyme. A recombinant baculovirus, encoding the human hepatoma cell (Hep G2) GGT, was easily purified using a histochemical procedure to reveal GGT activity. Infected insect cells synthesized a large amount of enzymatically active GGT representing up to 10% of the total cell extract protein. The GGT specific activity of the infected cells was 13 units per mg of protein which is the highest GGT expression level reported to date, 260-times more than in Hep G2 cells. The recombinant protein displayed an apparent molecular mass (M(r), 58,000 for the heavy subunit), immunoreactivity and catalytic features similar to those of the native protein. The high-level expression of functional GGT should provide an excellent tool to further study the structure-function relationships of the protein. PMID- 8728111 TI - EGF receptor in human Chang liver and hepatoma HepG2 cells. AB - Epidermal growth factor (EGF) receptor was detected in the human Chang liver and human hepatoma HepG2 cells. Both cell lines were found to be able to bind EGF. The expression of EGF receptor in Chang liver and HepG2 cells was 1.3 x 10(5) EGF receptors/cell and 1.8 x 10(5) EGF receptors/cell, respectively. In both cells, this receptor was identified by ECL Western blotting using monoclonal anti-EGF receptor antibody and by immunohistochemical assay using polyclonal anti-EGF receptor antibody. Some of internalized EGF was recycled in Chang liver cells, but not in HepG2 cells. PMID- 8728113 TI - Reductase and oxygenase activities of semisynthetic flavocytochromes based on cytochrome P450 2B4. AB - A synthetic flavocytochrome with the reductase and oxygenase activities was obtained by covalent binding of riboflavin to cytochrome P450 2B4. The reactions catalyzed by the newly synthesized flavocytochromes were studied. Formation of carboxycomplex with the reduced form of hemoprotein led to 60-80% inhibition of oxygenase reactions indicating the leading role of reduced heme iron in generating active oxygen species by flavocytochromes. PMID- 8728114 TI - A Fourier transform infrared spectroscopic study of the secondary structure of myelin basic protein in reconstituted myelin. AB - The secondary structure of myelin basic protein (MBP) in reconstituted central nervous system myelin was studied using Fourier transform infrared (FTIR) spectroscopy. The spectra of the protein in aqueous solution and in the lipid environment were compared and notable differences were observed. It is proposed that there are significant differences in the conformation of the protein in the contrasting environments. Significant increases in both alpha-helical structure and beta-structure were observed on reconstitution in myelin. The findings of this study also support the view that the presence of both alpha-helices and beta structure plays a important role in membrane proteins. PMID- 8728112 TI - Induction of DNA breaks in cardiac myoblast cells by norepinephrine. AB - We measured DNA single strand breaks (SSB) in cardiac myoblast cells in response to norepinephrine (NE) stimulation. Rat cardiac myoblast cells (H9c2) were stimulated with concentrations of 100 microMs to 1 mM NE for 2, 3, 4, and 12 hours after prior incubation with control solution, bunazosin, propranolol, verapamil, or captopril for 30 min. The DNA damage was measured by fluorometric alkaline elution. The strand scission factor, an index of the severity of SSB, increased slightly after stimulation with 200 microMs NE for 12 hours and with 1 mM NE for 4 hours. This increase was prevented by catalase or superoxide dismutase, which prevent production or accumulation of active oxygen radicals, during the stimulation, but not by pretreatment with a alpha-receptor antagonist, a beta-adrenergic receptor antagonist, a Ca2+ antagonist, or an angiotensin converting enzyme inhibitor. Thus, DNA SSB were induced by NE in cardiac myoblast cells. Certain active oxygen species may contribute to the DNA damage induced by NE. PMID- 8728115 TI - Apolipoprotein E is increased in aged rat kidney. AB - There is growing evidence to indicate that apolipoprotein (apo) E may be associated with age-related disorders and altered longevity in humans. Using rats as a model, we measured apoE in plasma, brain, heart, kidney, liver and spleen in aged (24-25 mo) and younger rats (6-8 mo). The results disclosed that: (a) the plasma concentrations of immunoreactive apoE in aged rats were higher than those in young animals by 70% (P < 0.01); (b) there was no age-related difference of apoE in the brain, heart, liver or spleen; (c) in contrast, the concentrations of apoE in the kidney of aged rats were markedly higher than those of young animals by 490% (P < 0.01). Our data suggest that, in the rat, age-related change in the organ concentrations of apoE is heterogeneous, and the selective increase in the kidney may have physiologic importance which merits further study. PMID- 8728116 TI - Biological activities of human tumor necrosis factor-alpha and its novel mutants. AB - Biological activities of human tumor necrosis factor-alpha (hTNF-alpha) and its mutants were compared. In cytotoxicity assay with L929 cells, one mutant, designated as TNF-B, showed 4.5-fold higher activity than TNF examined. And TNF-B also increased cytostatic activity and decreased protein synthesis against U937 cells. In receptor binding assay, TNF-B had almost the same affinity for TNF receports on L929 cells as hTNF-alpha. But another mutant, TNFarg, markedly decreased the all activities of hTNF-alpha and had lower affinity for receports on different types of target cell. These results indicated that the relative activity of TNFs to target cells may correlate with their affinity for receports. We also found that TNF-B retained the cytotoxicity of hTNF-alpha for HEp-2 cells. TNF-B also had two-fold higher affinity than hTNF-alpha for receptors on HEp-2 cells (only carrying hTNF-R55) and lower affinity for receptors on U937 cells (expressing mainly hTNF-R75). These results suggested that TNF-B might still interact with the human TNF-R55 receptor, but it might largely lose its ability to bind to human TNF-R75. Changes of biological activity of TNFs might be due to an altered affinity to the different types of TNF receptor on the target cells. PMID- 8728117 TI - Stimulation of mitochondrial oxygen consumption in isolated cardiomyocytes after hypoxia-reoxygenation. AB - An increase in mitochondrial matrix free calcium has been shown to occur during oxygen and substrate deprivation of the perfused heart which predisposes to calcium overload and inhibition of mitochondrial function on reoxygenation. In the current study we have assessed the effect of substrate free hypoxia on mitochondrial oxygen consumption and ATP synthesis in isolated rat cardiomyocytes. Myocytes were subjected to 40 min of substrate-free hypoxia and the oxygen consumption measured together with the effects on ATP and PCr synthesis. After hypoxia myocytes showed a fall in ATP to 10% of the control value. Within 5 sec of reoxygenation the ATP level recovered to a new steady state level of 30% of the original value. The rate of oxygen consumption of hypoxic/reoxygenated cells was 3-4 fold higher than that of cells maintained under normoxic controls but in the presence of oligomycin the difference was only 1.5-fold higher, indicating a greater requirement for mitochondrial synthesis of ATP following reoxygenation. Reoxygenation in the absence of extracellular Ca2+ resulted in a lower rate of oxygen consumption (50% of the rate measured in the presence of 1 mM-Ca2+) but did not affect the steady state concentration of ATP attained 5 min after reoxygenation. These results support the idea that the increased O2 consumption of myocytes following hypoxia/reoxygenation is due to an increased demand for ATP synthesis by mitochondria and is a response to the NA+ and Ca2+ loading of the cells which occurs under these conditions. This increased demand is likely to result in a greater generation of free radicals such as superoxide by the respiratory chain which could impair cellular function over the long term. PMID- 8728118 TI - Superoxide anion radical generation during the oxidation of various amines by diamine oxidase. AB - Diamine oxidase (DAO) or histaminase is an enzyme which deaminates histamine and several aliphatic amines to their corresponding aldehydes. Hydrogen peroxide and ammonia are side products of this reaction. The purpose of the present work was to evaluate if determination of produced hydrogen peroxide reflects DAO activity or if intermediate formation of the superoxide radical could be a reason for lack of correspondence between oxygen uptake and hydrogen peroxide production at different pH. Superoxide radical formation was determined by cytochrome c reduction in the presence and absence of superoxide dismutase (SOD). Oxygen uptake was measured with an oxygen electrode and hydrogen peroxide production by a spectrophotometric method. At pH 6.6 there was no superoxide production, but at pH 7.4 there was some, and it increased markedly at pH 9.5. Oxygen uptake also increased with increasing pH, especially with histamine as substrate. These results lead us to suggest that the mechanism of action of DAO involves the intermediate generation of superoxide radicals. PMID- 8728119 TI - Human cytochrome P450's are pro-oxidants in iron/ascorbate-initiated microsomal lipid peroxidation. AB - We have examined the effect of human cytochrome P450's (1A1,1A2,3A4,2A6,2B6,2D6,2E1) on ascorbate/iron-induced lipid peroxidation. Using microsomes prepared from human lymphoblastic cells enriched in recombinant cytochrome P450 isoenzymes, we have shown that the degree of peroxidation is a function of the amount of P450 present rather than the presence of any specific isoenzyme. Incorporated P450 increased the amount of peroxidation products by up to 2.1-fold compared to the control microsomes with no P450. It is therefore concluded that cytochrome P450's play a significant role in ascorbate/iron peroxidation. PMID- 8728120 TI - Reaction of ferrate (VI)/ferrate (V) with hydrogen peroxide and superoxide anion- a stopped-flow and premix pulse radiolysis study. AB - The reduction of ferrate(VI) to ferrate(V) by superoxide ions was studied over the pH range 2.6-13.0 using the premix pulse radiolysis technique. The pH dependence indicates that only the unstable protonated forms of ferrate, H2FeO4 (pKa3.5) and HFeO4- (pKa7.3) are reactive, k(HFeO4(-) + O2) = (1.7 +/- 0.2) x 10(7) M-1 s-1. The stable ferrate ion, FeO4(2-), showed no significant reactivity towards either hydrogen peroxide or superoxide anion. The rate constants for the spontaneous dimerization and decomposition of the protonated ferrates, e.g. k(HFeO4(-) + HFe04) approximately 250 M-1s-1, are orders of magnitude slower than their corresponding reduction reduction by superoxide indicating an outer-sphere mode of electron transfer for the latter process. In contrast the ferrate(VI) species H3FeO4+ (pKa = 1.6 +/- 0.2), H2FeO4, and HFeO4- oxidize hydrogen peroxide, e.g. k(HFeO4(-) + H2O2) = 170 M-1 s-1), at rates which correspond closely to their dimerization rates suggesting an inner-sphere controlled mechanism. PMID- 8728121 TI - Effect of radiation on red cell membrane and intracellular oxidative defense systems. AB - Ionizing radiation is currently used for prevention of transfusion associated graft versus host disease (TAGVHD). As radiation damage is associated with the production of activated oxygen species, the aim of this study was to observe the immediate effect of ionizing radiation on red cell membrane and intracellular oxidative defense systems. Neonatal and iron deficiency (IDA) cells, known for their increased sensitivity to oxidative stress, were chosen and compared with normal cells. Irradiation was performed in doses of 1500 cGy, 3000 cGy and 5000 cGy. GSH and methemoglobin levels and the activity of different antioxidant enzymes, measured under optimal in vitro conditions, were preserved in all cells after irradiation. Only radiation at the highest does of 5000 cGy, caused significant potassium leakage in neonatal cells and insignificant increase in IDA cells. Thus, cells with increased sensitivity to oxidative stress are more susceptible to damage by ionizing radiation than normal cells. PMID- 8728122 TI - Expression of bcl-2 inhibits cellular radical generation. AB - Bcl-2 expression in neural cells has been shown to inhibit apoptotic death in association with a decrease in reactive oxygen species. We present the results of a study that used electron spin resonance (ESR) measurements to evaluate the level of hydroxyl radical production in bcl-2 expressing GT1-7 cells and control cells. Incubation of cell monolayers with the spin trap N-t-alpha-phenylnitrone (PBN), and measurements of the hydroxyl radical production at different timepoints, revealed a higher radical production in control cells than in bcl-2 expressing cells, even in the absence of insult. The ESR signal was suppressed by addition of ethanol, indicating that the trapped radical was indeed hydroxyl radical. The mechanism by which the expression of bcl-2 leads to a decrease in cellular production of hydroxyl radical is unknown. PMID- 8728123 TI - Effects of controlled exposure of sunlight on plasma and skin levels of beta carotene. AB - We conducted a randomized placebo-controlled double-blind study in 20 healthy young female students (skin type II + III, body mass index 18-22) in order to evaluate the efficacy of 10 weeks of moderate dose (30 mg/d) beta-carotene (BC) on plasma and skin beta-carotene levels during 12 days of time and intensity controlled sunlight exposure at sea level (30 degrees latitude, Red Sea, Eilath, Israel). After 12 days of controlled sun exposure (total UV dose of about 10.000J/cm2), plasma beta-carotene decreased in the placebo (p < 0.01) and beta carotene group (not significant). In addition cutaneous beta-carotene decreased significantly in both groups. Plasma alpha-tocopherol decreased significantly (p < 0.01) during exposure time in both groups. In the supplemented group, however, the decrease of a-tocopherol was significantly greater (p < 0.01) than in the placebo group. We conclude that sunlight influences the beta-carotene and alpha tocopherol content of blood and tissues. PMID- 8728124 TI - Oxidation of caffeine and related methylxanthines in ascorbate and polyphenol driven Fenton-type oxidations. AB - Caffeine and related methylxanthines were subjected to free radical mediated oxidation by incubation with Fe(3+)-EDTA/ascorbate and Fe(3+)-EDTA/polyphenolics. The reaction mixtures were analysed by reverse-phase HPLC, revealing the corresponding C-8 hydroxylated analogues as the major products of hydroxyl radical mediated attack. Further oxidation products of caffeine, analysed by liquid chromatography-mass spectrometry (LC-MS), were the N1-, N3- and N7 demethylated methylxanthine analogues theobromine, paraxanthine and theophylline, respectively. Isolable amounts of the imidazole ring operated 6-amino-5-(N formylmethyl-amino)-1,3-dimethyl-uracil (1,3,7-DAU) derivative were also detected, which was characterised by 1H NMR and mass spectroscopy. The identified products indicate that the pertinent chemical reactions, i.e. C-8 hydroxylation, demethylations, and C8-N9 bond scission, are comparable to the primary metabolic pathways of caffeine in humans. The influence of pH, transition metals, hydrogen peroxide, free radical scavengers and metal chelators on caffeine oxidation was studied. This report illustrates that natural food-borne reactants can aid in identifying specific chemical markers of free radical induced damage. Furthermore, potentially anti-and pro-oxidative reactions can be elucidated which may be important in assessing the impact of nutrient additives and supplements on the shelf life and stability of foods and beverages. PMID- 8728125 TI - Intervention research: science, skills, and strategies. AB - Despite a rich history of etiological research, the field of occupational safety and health does not have a rigorous history of research on what works and does not work to prevent and control occupational diseases and injuries. National and global transformations of economies and workplaces with enhanced competitiveness require more attention to options for interventions. A three-pronged approach to building a body of knowledge on intervention research in occupational health and safety is identified in this paper. The approach focuses on the science, skills, and strategies that can be useful in intervention research. Scientifically, researchers can draw on constructs and techniques from epidemiology, evaluation practice, and clinical trials. Experimental and nonexperimental approaches have value for occupational studies. The skills needed represent a range of disciplines beyond those traditional of health and safety; social scientists, economists, and organizational theorists often need to be part of research teams. Strategic approaches involve more labor-management partnerships, prospective study designs, and the use of intermediate and surrogate indicators. The strategic challenge will be to conduct intervention research against a backdrop of overriding political and economic pressures. PMID- 8728126 TI - Methodological issues for intervention research in occupational health and safety. AB - This article presents a brief summary of the nature and extent of intervention research being conducted in the area of occupational safety and health. Articles were classified either as engineering, administrative, or behavioral, according to the type(s) of interventions that were evaluated. Findings suggest that many of the intervention studies conducted lacked a theoretical basis, used small samples, and tested interventions lacking the intensity to cause the desired change. Most study designs were either nonexperimental or quasi-experimental. Recommendations for conducting future research are presented. PMID- 8728127 TI - The future of intervention research at NIOSH. AB - The National Institute for Occupational Safety and Health (NIOSH) has recently made a commitment to increase both extramural and intramural support of control technology and intervention research. It is important for NIOSH to use intervention research more aggressively because it provides a mechanism to go beyond investigation, identification, and recommendations to actually determine if prevention has occurred. Intervention research can assess the effectiveness of the hierarchy of controls, workplace standards, and health and medical care, as well as provide important information on occupational disease and injury surveillance and health communication efforts. In pursuing intervention research, NIOSH will focus on enhancing its existing control technology and surveillance programs, seeking input from partners in industry and labor, pursuing interdisciplinary approaches, considering the cost and feasibility of controls, considering and integrating behavioral procedures, and widely disseminating the results. PMID- 8728128 TI - Key issues in intervention research: a program evaluation perspective. AB - Over the last two decades, theory and practice in the field of program evaluation have generated a rich array of concepts and methods for research on the effectiveness of social programs. This paper attempts to summarize the lessons from program evaluation research that might usefully inform intervention research in occupational health and safety. PMID- 8728129 TI - Engineering controls as an intervention to reduce worker exposure. AB - The implementation of controls to reduce worker exposure should be considered the ultimate goal of any successful industrial hygiene program. The industrial hygiene literature has consistently described a hierarchy of controls, consisting first of the engineering controls (substitution, isolation, ventilation), and followed by administrative controls (personal protective equipment, worker education, scheduling etc.). Recently, exhaust ventilation has been the most popular form of engineering control technology for controlling exposure to airborne contaminants. The use of ventilation to control exposures is not without its problems, however, and many of these problems potentially are more severe in smaller companies. This paper proposes a new emphasis on the first control in the hierarchy, substitution. Historically, substitution has meant the substitution of a hazardous chemical or process by one that is less so. This definition is too restrictive; because of this, it is proposed instead to use the term process change, defined as the use of any process modifications that serve to reduce worker exposure. The advantages and disadvantages of the process change approach are discussed and are illustrated with case studies. PMID- 8728130 TI - Psychosocial aspects of interventions in occupational safety and health. AB - Attention to psychosocial hazards in work environments should become an increasingly important component of occupational safety and health interventions. Research findings have linked a number of measurable psychosocial characteristics with negative psychological and physiological consequences. Some serious physical health hazards have also been found to have psychosocial components. Several economic trends indicate that more jobs will be created with high levels of psychosocial hazards. These economic trends include a decline in wages, a move away from a manufacturing base, increased hours of work, a decline in unionization rates, and poorly implemented technological changes. Intervention strategies developed to reduce psychosocial stressors must address the multiple cause of psychosocial hazards and the multiple symptoms they produce. Additional challenges include assessing psychosocial changes within the sometimes contradictory framework of organizational and social changes. Nevertheless, comprehensive efforts to improve both the health and safety and the overall quality of working life need to incorporate psychosocial variables into their designs. PMID- 8728131 TI - Intervention research in occupational safety and health: examples from construction. AB - Construction is one of the largest industries in the United States, with 13% of the gross national product and 5-6% of the labor force. It is also one of the most dangerous industries, accounting for 15% of occupational fatalities and 17% of all workers' compensation costs. In 1989, the Building and Construction Trades Department, AFL-CIO, completed an agreement with the National Institute for Occupational Safety and Health to develop a national labor-management initiative to improve occupational safety and health throughout the construction industry. The aim was to remedy a lack of research on construction occupational safety and health. The first years were spent on surveillance to characterize construction safety and health problems, development of awareness about safety and health issues among decisionmakers in the industry, and some limited interventions. A second phase was initiated in 1994, which focuses on intervention activities. Results from this joint program include a growth in annual federal construction safety and health research expenditure from $300,000 in 1989 to $12 million in 1995, a research network that now encompasses more than 30 institutions, a national conference that established an agenda to change construction safety and health, four regional conferences to develop coalitions and implementation strategies, and the development of a feasible goal to reduce fatality and injury rates by 80%. The program may already be having an impact. According to the Bureau of Labor Statistics, lost-time injury rates for construction for the three most recent years of reporting declined by 20%. PMID- 8728132 TI - The role of surveillance in the hierarchy of prevention. AB - Surveillance is the collection, analysis, and dissemination of results for the purpose of prevention. Surveillance tells us what our problems are, how big they are, where the solutions should be directed, how well (or poorly) our solutions have worked, and if, over time, there is improvement or deterioration. Surveillance is essential to successful sustained public health intervention for the purposes of prevention. Surveillance systems must be tailored to the specific disease or injury that is to be prevented. Surveillance should not be limited to the occurrence of death, disease, or disability. Public health is a multilevel cascade of activities involving recognition, evaluation, and intervention. Public health should include elements of experimentation as well as field implementation with evaluation. Surveillance is the mechanism to modify any element in the cascade based upon that element's contribution to prevention or lack thereof. Any element in the causal or intervention pathway is appropriate for surveillance as long as the monitoring of the element is useful in improving the prevention system. These elements include the occurrence of hazard and intervention as well as disease, death, or disability. Examples will be provided that demonstrate the roles of surveillance in the recognition of new diseases, the evaluation of the persistence of recognized problems, the estimation of the magnitude and trends of public health problems, and the provision of information to motivate intervention. PMID- 8728133 TI - Intervention research: a model from the National Cancer Institute's Smoking and Tobacco Control Program. AB - Recognizing that the scientific method is as critical to cancer control as it is to basic laboratory research, the National Cancer Institute (NCI) established a well-defined, systematic strategy for attaining its cancer control goals and objectives. This strategy, operationalized in the early 1980s as a five-phase process, emphasized cancer control as a research science rather than a demonstration science. The five phases of NCI's cancer control research strategy progress from hypothesis development, to methods development, to controlled intervention trials, to defined population studies, and finally to demonstration and implementation programs. This research base provides the foundation for nationwide prevention and health services programs. The application of this five phase approach to NCI's efforts to reduce morbidity and mortality attributable to tobacco use is described, and some of the challenges that faced the Institute in this process are identified. These experiences provide an important framework for other disciplines faced with the challenge of translating science into practice. PMID- 8728134 TI - Qualitative methods for intervention research. AB - Intervention research takes place in field settings and requires understanding of social meanings and social processes. These are tasks for which qualitative research methods are well suited. The purpose of this paper is to provide a starting point for those who would like to learn more about the qualitative research methods used in disciplines where the study of social phenomena in naturalistic settings is common-particularly sociology, cultural anthropology, and human services program evaluation. The paper discusses some ways that qualitative and quantitative methodologies can usefully work together, outlines four key differences in the initial premises of quantitative and qualitative research approaches, briefly reviews some methodological techniques useful in gathering and analyzing qualitative data, and provides suggestions for further reading on various aspects of qualitative research. PMID- 8728135 TI - Occupational stress intervention. AB - The topic of occupational stress has received considerable research attention during the last decade and has emerged as an important occupational safety and health concern. Worker compensation claims for stress-related illnesses, for example, were the fastest growing type of claim in the 1980s, comprising more that 11% of all such claims. Concern over problems associated with occupational stress and their costs has fostered interest in intervention strategies. While specific work stressors and their resulting physical and mental health consequences have been identified, relatively few successful interventions have been documented in the literature. This article discusses primary, secondary, and tertiary interventions efforts in the area of occupational stress and argues for efforts to increase understanding of the occupational stress intervention process. PMID- 8728136 TI - An improved model for professional conferences and workshops. AB - A cooperative learning method designed to increase learning and other benefits to participants is presented. Small group problem-solving from case studies or story narratives provides a way to overcome the limitations of conventional lecture presentations. An example that used a case study in small groups during a breakout session at a scientific conference to improve skills in intervention design and research evaluation is described. PMID- 8728137 TI - Policy evaluation: better living through research. AB - Public health scientists have produced valuable research about the epidemiology of occupational hazards, their measurement, and engineering controls. Still, many firms do not apply available knowledge to eliminate workplace hazards. Occupational safety and health policy research helps to bridge the gap between current scientific understanding and effective public policies. It focuses on four areas: (1) primary prevention policies, including standards focusing on controlling specific hazards, standards requiring health and safety programs, surveillance, education and training, targeting of enforcement, and nonregulatory safety incentives; (2) secondary prevention policies, including medical care, vocational rehabilitation, and laws and regulations fostering reemployment of injured workers; (3) compensation policies, including the range and level of medical benefits and income benefits to injured workers; and (4) behavioral responses to policies that lead to unintended consequences. This article provides examples of existing research in these areas and discusses the direction of future occupational safety and health policy evaluation research. PMID- 8728138 TI - Approaches for assessing the efficacy of occupational health and safety standards. AB - The regulation of hazards is one of the most dramatic forms of intervention in occupational safety and health (OSH). Despite their high degree of potential social and economic impact, relatively little research has been conducted to specifically evaluate the effectiveness of OSH standards with regard to preventing occupational diseases and injuries. This paper reviews the basic scientific approaches that may be used to evaluate the efficacy of OSH standards. These approaches encompass the following research areas: (1) exposure surveillance, (2) disease surveillance, and (3) prospective studies following the introduction of the standard. Research on asbestos and asbestosis, respirable crystalline silica (quartz) and silicosis, and respirable coal mine dust and coal workers' pneumoconiosis (CWP) are used to illustrate these approaches and the type of information that is currently available. The examples (quartz, coal dust, asbestos) reveal substantial limitations in the types of information currently available for evaluating the efficacy of these OSH standards. Ideally, plans for evaluating the efficacy of OSH standards should be developed for existing and future standards. These plans should include programs for the surveillance of exposures and adverse health effects and, when possible, for prospective studies designed to evaluate how the risk of disease (or injury) is modified by the introduction of the standard. PMID- 8728139 TI - Policy implications of intervention research: research on the social context for intervention. AB - Intervention research is of significance because the failure or success of particular interventions may have influence on policy and practice. Poorly designed interventions may impede progress in health and safety. Frequently, interventions are based on scientific/ technical definitions of occupational health and safety problems but involve intrusion into complex sociotechnical work environments. Intervention research must be built on a solid foundation of social science research. An example of research useful to health and safety intervention is the Gray Institute study of management practices in the chemical industry that was commissioned by OSHA a few years ago. It focused on safety management in chemical manufacturing firms and their contractors, including a survey of managers and contractors and nine intensive case studies of particular plants. The study describes safety management practices in the best firms in the industry. Such research can provide the groundwork for effective safety interventions in the industry, including training and training requirements, experimentation with redefined managerial responsibilities, regulation of contractors, and others. It also suggests the kinds of useful data that might be collected by NIOSH in the normal course of health and safety studies. Finally, it suggests that the interdisciplinary teams currently investigating occupational health and safety should be expanded to include social scientists. PMID- 8728141 TI - Overcoming practical challenges in intervention research in occupational health and safety. AB - In addition to more familiar research issues, intervention research projects in naturalistic settings present the investigator with a number of practical challenges including gaining access to potentially resistant populations, maximizing participation rates in the face of weak incentives for cooperation, getting valid answers to sensitive questions, and meeting ethical obligations when health or legal problems are discovered in the course of study. Generalizable approaches to these challenges are addressed in the context of a retrospective evaluation of the implementation of OSHA's 1984 ethylene oxide standard in Massachusetts hospitals. In the evaluation study, enthusiastic cooperation was secured, a 96% participation rate was realized, sensitive questions were posed successfully, and worker health risks discovered in the course of study received attention without having to wait for the write-up of the study results. Key elements in the study population's receptivity appear to have been (I) the investigator's familiarity with the hazard, its setting, respondent concerns and needs, and (2) reciprocity in the form of providing follow-up consulting services as an integral part of the research process, delivered to each hospital at the conclusion of data collection. Specific techniques used in the study are presented in the hopes of aiding other investigators facing similar practical challenges in occupational health and safety intervention research. PMID- 8728140 TI - Intervention research: GAO experiences. AB - This paper describes tools of program evaluation that may prove useful in conducting research on occupational health and safety interventions. It presents examples of three studies conducted by the U.S. General Accounting Office that illustrate a variety of techniques for collecting and analyzing data on program interventions, including analysis of extant data, synthesis of results of existing studies, and combining data from administrative files with survey results. At the same time, it stresses the importance and difficulty of constructing an adequate "theory" of how the intervention is expected to affect outcomes, both for guiding data collection and for allowing adequate interpretation of results. PMID- 8728143 TI - Etiologic and prevention effectiveness intervention studies in occupational health. AB - In this article a basic distinction is made between etiologic and prevention effectiveness intervention studies. Etiologic intervention studies focus on elucidating causes of disease, while the purpose of prevention effectiveness intervention studies is to study methods of prevention. The design requirements for each of these studies are very different: etiologic intervention studies usually need large study populations, large exposure contrasts, ascertainment of exposure, as well as health outcome. Ideally, randomization and blinding should also be applied. Effective preventive strategies may, on the other hand, be identified in small study populations with exposure as the only outcome measure, and randomization and blinding may be superfluous. At present, intervention studies are in great demand, and often there is a wish that etiologic questions as well as prevention effectiveness be addressed in the same study. We argue that this should not be done without careful consideration of possible conflicting design aspects. PMID- 8728142 TI - Making the "new American workplace" safe and healthy: a joint labor-management researcher approach. AB - The American workplace is now in the midst of the most significant change since the advent of mass production. Whether these changes will lead to improvements in worker health and safety is not clear. This paper describes an approach to intervention and research-participatory action research (PAR)-that has the potential to redesign work organizations to improve performance while also improving health and safety. In the PAR method, researchers, managers, workers, and unions collaborate in a process of data-guided problem solving intended both to improve the system's performance and to contribute to general scientific knowledge. A case study example illustrates the use of a PAR approach in an automobile parts facility where labor, management, and researchers jointly conducted a longitudinal project aimed at reducing the major sources of stress and enhancing employee well-being. Results from the 6 year project suggest that, properly implemented, PAR has the potential to both lead to improved intervention and contribute to theoretical advances in occupational safety and health. The PAR approach to intervention research is contrasted with the total quality approach (TQA), and some suggestions are made for improving PAR research designs. PMID- 8728144 TI - An engineer's perspective of the intervention research workshop. AB - As an engineer practicing in the area of engineering controls for the reduction of occupational exposures to hazardous agents, the Intervention Research Workshop studies appeared to be effective for the studied situations, yet the models may not be applicable across a wide range of intervention possibilities. A more generic model might be more appropriate, perhaps a business model. Dr. W. Edwards Deming has proposed a model for the production of goods and services. If we can view interventions as products which need to be sold to potential customers, perhaps this type of model would be more effective across a wide range of intervention strategies. PMID- 8728145 TI - Intervention research in agriculture: examples from the swine confinement and respiratory health project. AB - The "Measurement and Analysis in Agricultural Interventions" workshop session of the "Intervention Research in Occupational Health and Safety: Science, Skills, Strategies" conference considered a variety of approaches to safety and health interventions in agriculture. The "Respiratory Health in Swine Confinement Project" in Iowa, an educational intervention to improve respiratory health and safety in swine confinement workers, was presented as a case study for the discussion. Results from the project were used to illustrate the advantages and disadvantages of a variety of research techniques in interventions and program evaluation, including specific issues related to measurement and analysis. The discussion reflects comments from workshop participants along with summary observations reported to conference attendees. Themes of the session include the complementary nature of quantitative and qualitative techniques, and the importance of developing interventions that are community based. PMID- 8728146 TI - An occupational health and safety intervention research agenda for production agriculture: does safety education work? AB - It is clear that agriculture has not kept pace with other hazardous industries in reducing its injury rate. For example, between 1960 and 1990 the work death rate for agriculture decreased only 28% while the work death rates decreased 65% for mining and 55% for construction [Purschwitz (1992)]. A national conference in Iowa in 1988 came to the forceful conclusion that "America's most productive workforce was being systematically liquidated by an epidemic of occupational disease and traumatic death and injury" [NCASH (1988)]. In 1991, the nation's top public health officer, the U.S. Surgeon General, convened a conference titled "FarmSafe 2000-A National Coalition for Local Action," to formally address agricultural safety and health issues. Importantly, conferees recognized that preventing injury and disease was superior to trying to rehabilitate people after an injury occurred. But does participation in farm safety and health educational programs lead to a reduction in risk of injury from farm work? Questions are being raised about the value of farm safety and health educational information, campaigns, programs, and related activities. The questions have identified a critical gap in the literature of farm safety and health education. There is currently no good evidence demonstrating that farm safety and health education, campaigns, programs, or related activities lead to a relatively stable reduction of risk on the farm. In other words, do farmers and their families actually put to use, in a relatively permanent or stable manner, the educational information regarding elimination, reduction, or control of physical hazards and the modification of work behavior that may cause injury? PMID- 8728147 TI - Machining operations and associated machining fluid exposures: issues for health and safety intervention in manufacturing. AB - The outline for a workshop discussion of intervention research in manufacturing industry is presented. The example used is the control of machining fluid exposures, a widespread exposure that is associated with risk of cancer, pulmonary disease, dermatitis, and several types of accident-related injuries. Relevant technical considerations for conducting intervention research include the selection of machining fluid for different machining purposes, the management of fluids in use, and the selection and maintenance of environmental controls. Equally important are nontechnical considerations including the need for creating labor-management health and safety committees, the use of administrative rules on job performance for both workers and management, and training on the role of individuals from either group in exposure control. Clarity concerning the objectives for an intervention study and plans for clear means to evaluate study outcomes is critical. Important study variables will include site selection, plant size, plant demographics, the involvement of workers in the study, a willing management, and concern for susceptible workers. PMID- 8728148 TI - Use of participatory ergonomics teams to address musculoskeletal hazards in the red meat packing industry. AB - Participatory ergonomics teams from two departments of a pork slaughtering plant analyzed musculoskeletal hazards and proposed ergonomics solutions for several jobs in the plant. The jobs were targeted based on association with a large number of injuries, one or more particularly severe injuries, high workers' compensation expenses, and/or high turnover plus absence of obvious solutions to abate the hazards. The teams used a structured problem-solving method that was generally derived from principles associated with quality improvement processes. Workers performing the targeted jobs were involved in the problem-solving process, even if they were not members of the team. The effectiveness of each team was assessed and compared in terms of the number of problem jobs addressed and feedback from team members. One team addressed all nine of its targeted jobs; the other team addressed eight of 12 targeted jobs. Feedback from team members regarding their teams' methods and performance was generally favorable. The major obstacle to effective team functioning in this project was the scheduling of meetings. There were also some obstacles related to team leadership. Overall, this project demonstrated that the use of participatory ergonomics teams that rely on structured problem-solving methods are able to work effectively to address musculoskeletal hazards, especially related to the upper extremities, in the red meat packing industry. PMID- 8728149 TI - Encouraging small businesses to adopt effective technologies to prevent exposure to health hazards. AB - Small businesses are heterogeneous and the prospects are low for direct OSHA inspection and enforcement. Opportunities are explored to encourage voluntary adoption of new technology to reduce workplace exposures. The case of radiator repair shops is used in this paper to illustrate an approach to the dissemination of control technology to small businesses that will encourage these companies to adopt controls. Several behavioral theories are applied to the case. PMID- 8728150 TI - Methodologic issues in intervention research--health care. AB - To better understand the methodological challenges faced by intervention research in health care, workshops reviewed two intervention studies to reduce back injuries among nursing home staff and two studies on the use of precautions to prevent occupational transmission of bloodborne pathogens. These studies adapted rigorous designs to real-world settings and made good use of multiple measures to detect effects and communicate this information to policy makers. The studies grappled with issues about implementation integrity and would benefit from better theory of administrative practices associated with a safety-conscious work environment. PMID- 8728151 TI - The role of cues in intervention research: a query. PMID- 8728152 TI - Facilitator report: bloodborne pathogens exposure among health care workers. AB - Several psychosocial and organizational factors have been identified as important correlates of compliance with safe work practices among health care workers. In particular, compliance with "universal precautions"--a set of work practices which were designed to minimize exposure to bloodborne pathogens--has been found to be associated with several potentially modifiable factors. The development of interventional strategies designed specifically to target these factors (e.g., HIV/AIDS attitudes, safety climate) are important objectives in the overall risk management approach to occupational exposure to bloodborne pathogens. PMID- 8728153 TI - NIOSH research initiatives to prevent back injuries to nursing assistants, aides, and orderlies in nursing homes. AB - Over the past 100 years, advances in nutrition, modern medicine, public health, and a multitude of public health improvements have increased the life expectancy of U.S. residents. The fact that Americans are living longer has resulted in extensive growth in our elderly population and a rapid employment growth that delivered about 2 million new jobs between 1980 and 1989 in the health care workforce. The Bureau of Labor Statistics Injury and Illness Data for nursing homes rose from 10.7 to 18.6 injuries or illnesses per 100 full-time workers between 1980 and 1992. The injury and illness rates among nursing home workers are partly due to the physical stress of providing round-the-clock assistance with the basic activities of daily living, such as getting in and out of a bed or chair, as well as bathing and toileting. The National Institute for Occupational Safety and Health (NIOSH) is conducting a series of research studies to identify strategies to reduce the risk of musculoskeletal injuries to workers in nursing homes. NIOSH has funded two laboratory evaluations of resident transferring methods and one field study in an actual nursing home. The purpose of this paper is to describe the key findings from past NIOSH research initiatives and to present an overview of future research. PMID- 8728154 TI - Emerging participatory approaches to ergonomic interventions in the construction industry. AB - The Central Artery/Third Harbor Tunnel (CA/T) construction project in Boston is the largest and most costly highway construction project ever undertaken in the United States. The Construction Occupational Health Project (COHP) of the Work Environment Department at the University of Massachusetts-Lowell was established to conduct exposure assessment and to develop, introduce, and evaluate interventions in the areas of ergonomics and industrial hygiene on the CA/T project. For both political and practical reasons, COHP is using a participatory approach to intervention in the construction industry. The research process is employing participation at all the levels of the construction hierarchy in the form of various advisory groups. These advisory groups have been formed from existing joint labor-management advisory committees and are presently engaged in two participatory intervention activities: (1) evaluations of intervention ideas, and (2) comparisons of safety systems. PMID- 8728155 TI - Intervention research in construction: a hypothetical case study of painters. AB - This paper presents a theory on which to base intervention research in the construction industry, along with a hypothetical case study involving construction painters. The hypothetical case is based on data from research in a three-state study of construction painters currently under way. Difficulties facing researchers who conduct research in construction are described. Advantages of the intervention research design for construction are discussed. PMID- 8728156 TI - CAPD overview. PMID- 8728157 TI - Cardiovascular disease in peritoneal dialysis. AB - Cardiovascular morbidity and mortality remain high in ESRD patients. Lipid abnormalities in CAPD may be more important than in hemodialysis. Vessel calcification may have a role in atherosclerotic heart disease, but this is only an inference from several clinical observations, and it remains to be defined more clearly as a risk factor. Left ventricular hypertrophy is frequent in this patient population, and is associated with specific clinical patterns and an increased risk of death. Erythropoietin treatment of anemia and tight blood pressure controls have proved to help in reversing severe left ventricular hypertrophy. Finally, we describe a syndrome of the hypertrophic, high cardiac output hemodialysis heart, which is characterized by a high cardiac output in hemodialysis patients. It is associated with left ventricular hypertrophy and eventually right ventricular hypertrophy with tricuspid insufficiency. This may require fistula revision and even a switch peritoneal dialysis. PMID- 8728158 TI - Aquaporin water channels in the kidney: localization and regulation. PMID- 8728159 TI - New strategies in clinical nutrition. PMID- 8728160 TI - Pathogenesis of peritoneal fibrosis and peritoneal small vessel changes. PMID- 8728161 TI - Markers of peritoneal membrane status. PMID- 8728162 TI - Advanced glycation end products in peritoneal dialysis. PMID- 8728163 TI - Demonstration of aquaporin-CHIP in peritoneal tissue of uremic and CAPD patients. AB - Aquaporin-CHIP is a 28 kD channel forming integral membrane protein. It acts as an osmotically driven, water-selective pore. The presence of aquaporin-CHIP has been demonstrated in the proximal tubule in the kidney and in the pleura, as well as in other tissues. During peritoneal dialysis a dissociation between the transport of water and sodium using hyperosmolar solutions has been reported, suggesting the presence of ultrasmall pores. Water channels, like aquaporin-CHIP, could be the morphological equivalent of these pores. We investigated the possible presence of aquaporin-CHIP in cryo-sections of peritoneal tissue using affinity purified human anti-CHIP IgC (P. Agre, Baltimore, MD). Peritoneal biopsies (omenta) were obtained at catheter insertion in 2 uremic patients with end-stage renal disease, and at catheter reimplantation of 1 patient treated with continuous ambulatory peritoneal dialysis (CAPD) for two years. Peritoneal tissue obtained at autopsy from 1 patient who had been on CAPD for four years, but in whom CAPD had been discontinued for five months, was also studied. Aquaporin-CHIP antiserum specific staining was found in the endothelial cells of the peritoneal capillaries in all patients. No obvious difference in the intensity of staining was seen between uremic and CAPD patients. This demonstration of aquaporin-CHIP in human peritoneal endothelial cells supports the hypothesis of the existence of ultrasmall pores within the peritoneal membrane. These water channels facilitate the transcellular transport of water, induced by an osmotic gradient, in the absence of sodium transport. It may be the explanation for the dissociation of water and sodium transport that occurs during hyperosmolar solutions. Aquaporin CHIP is present in human peritoneal endothelial cells in both uremic and CAPD patients. Aquaporin-CHIP may be the morphological equivalent of the ultrasmall pores within the peritoneal membrane. PMID- 8728164 TI - Interleukin-1 beta stimulates glucose uptake of human peritoneal mesothelial cells in vitro. AB - To investigate whether the glucose uptake (GU) of human peritoneal mesothelial cells (HPMC) is mediated by glucose transporters and whether this uptake is influenced by interleukin 1-beta (IL-1 beta), we measured 2-deoxy-(3H)-GU of HPMC in vitro, after exposing the cells for different times (two and 12 hours) to increasing concentrations (0.1, 1.0, and 2.0 ng/mL) of IL-1 beta. To exclude a noncarrier-mediated transport, GU was also tested in the presence of cytochalasin B. All experiments were performed in triplicate in the cells of two donors. Cytochalasin B inhibits GU of HPMC almost completely. GU of HPMC is not stimulated by insulin. GU is stimulated by IL-1 beta in a dose-dependent manner. These data indicate a GU of HPMC, which is mediated by a glucose transporter and stimulated by IL-1 beta. The increased uptake of glucose from the dialysate in patients with peritonitis may be mediated by a (cytokine-induced) increased activity of HPMC glucose transporters. PMID- 8728165 TI - Paradoxes in peritoneal transport of small solutes. AB - Analysis of kinetic studies of peritoneal solute transport involves the need for discrimination between three transport components: diffusion, convective transport, and peritoneal absorption. The description of convective transport in standard clinical conditions of continuous ambulatory peritoneal dialysis (CAPD), as well as in isochratic measurements, has met some problems related to the paradoxical and often anomalous values of sieving coefficient, a parameter that characterizes solute drag with the flow of ultrafiltrate. A possible explanation of some of these results is the time dependence of the transport parameters, which is in contrast to their assumed steadiness. These anomalies as well as the time dependence of the transport parameters are confined more to the standard glucose-based dialysis fluid than to some alternative dialysis fluids. Furthermore, the most striking anomalies have been found for small electrolytes as well as for osmotic agents, which are applied in high, unphysiological concentrations. These solutes may be involved in the transport between intracellular and extracellular compartments within the peritoneal membrane, which phenomena are not included in the current modeling. PMID- 8728166 TI - Peritoneal blood flow: does it matter? PMID- 8728167 TI - The importance of the interstitium in peritoneal transport. AB - The peritoneal capillary exchange vessels are located within all the tissues which surround the peritoneal cavity and are separated from the peritoneal dialysis fluid by the tissue interstitium. The interstitium adds an additional barrier to transcapillary transport resistance and slows the diffusion of solutes from the blood to the dialysis fluid. The interstitium also alters the pressure environment of the blood capillary and has profound effects on water transport, causing fluid loss from the cavity to the body during dialysis. PMID- 8728168 TI - Role of diaphragmatic, visceral, and parietal pathways in peritoneal fluid absorption in rat peritoneal dialysis. AB - Assessment was made of the contribution of lymphatic and nonlymphatic fluid absorption to net fluid loss from the peritoneal cavity. Diaphragmatic, visceral, and parietal pathways in lymphatics and nonlymphatics were examined using a rat model with adhesion of the diaphragm to the liver, evisceration, these two procedures in combination, and without treatment. In each of these cases, six rats were used, each dialyzed for 180 min with Krebs-Ringer solution. The peritoneal net fluid absorption rate (PNFAR) was determined based on the disappearance of 125I-bovine serum albumin (BSA) from the peritoneal cavity and the lymphatic absorption rate (LAR), was based on the appearance of this albumin in the blood. Seventy-eight percent of net fluid loss occurred via the nonlymphatic pathway, primarily through parietal and visceral absorption; and the remaining 22% through the lymphatics, the main pathway being the subdiaphragmatic lymphatics. Nonlymphatic fluid absorption would thus appear to be a major route of fluid loss from the peritoneal cavity in rat peritoneal dialysis. PMID- 8728169 TI - Measurement of hydrostatic intraperitoneal pressure: a necessary routine test in peritoneal dialysis. AB - This paper summarizes our clinical studies on hydrostatic intraperitoneal pressure (IPP), showing the interest of this measurement in routine clinical practice. IPP can easily be measured routinely be a simple and safe method: the measure of the column of dialysate in the peritoneal dialysis (PD) line before drainage, with point 0 located on the midaxillary line. The normal value is 12 +/ 2 cm of water (cm H2O) with an intraperitoneal volume (IPV) of 2 L, with linear increases of 2.2 cm H2O for each additional liter. IPP must be measured to estimate the tolerance of IPV: the maximal permissible IPV is reached for an IPP of 18 cm H2O, squaring with a decrease of 20% in vital capacity and sometimes arising before clinical symptomatology. However, IPP measured at rest could not predict PD mechanical complications (hernias, dialysis leakages, hemorrhoids, etc.), which are more dependent on parietal previous history or predisposition. IPP is significantly higher during the first three days after peritoneal catheter implantation (17 +/- 3 cm H2O) than during the 12 following days (10 +/- 4 cm H2O). It is recommended to postpone the start of PD until after catheter implantation, and patients should remain supine for the first three days. On the other hand, IPP strongly reduces the overall ultrafiltration (UF) volume: an increase of 1 cm H2O in IPP caused a decrease of 70 mL in global UF after two hours. Therefore, IPP should be measured in diagnosis of losses of UF. However, UF loss during peritonitis is not due to an increase of IPP. PMID- 8728170 TI - Charge-affected transperitoneal movement of amino acids in CAPD. AB - Our objective was to investigate the influence of molecular charge on transperitoneal solute movement in continuous ambulatory peritoneal dialysis (CAPD). Tests of peritoneal equilibration were performed. Two liters of 2.27% or 2.5% glucose CAPD dialysate were infused and the dialysate samples were taken after 2 hr and 4 hr, and blood samples were obtained after 4-hr dwell time. Dialysate-to-plasma concentrations ratios (D/P) were calculated for creatinine (Cr) and three amino acids with almost the same molecular weight but quite different charges: glutamic acids (Glu: negatively charged), glutamine (Gln: near neutrally charged), and lysine (Lys: positively charged). The setting was a university hospital. There were 23 stable CAPD patients with a mean age of 56.6 +/- 9.5 years and a mean CAPD duration of 15.2 +/- 19.4 months. D/P ratio of Glu was much lower than those of Gln, Lys and Cr at both 2 hr and 4 hr (p < 0.01), and D/P of Lys was significantly lower than that of Gln (p < 0.01). There was no significant difference of D/P between Gln and Cr. The order of transperitoneal mobility among the three amino acids was Gln > Lys > Glu. Transperitoneal movement of solutes in CAPD is influenced by molecular charge, the movement of negatively charged solutes is most remarkably retarded in cases of amino acids. PMID- 8728171 TI - Minoxidil selectively improves peritoneal ultrafiltration. AB - This cross-over randomized clinical trial was carried out to evaluate the effects of intraperitoneal (i.p.) administration of minoxidil on fluid removal and solute clearance during peritoneal dialysis. Twenty-one patients with endstage renal disease, awaiting enrollment in chronic hemodialysis therapy, were randomly allocated to receive i.p. minoxidil either in the first or the last nine cycles of a 24-cycle peritoneal dialysis session. Cycle-to-cycle data on fluid balance, blood pressure, and adverse effects of the drug were obtained. The dialysis fluid recovered in cycles 3, 6, 9, 18, 21, and 24 was analyzed, together with plasma, for creatinine, urea nitrogen, and protein content. The mean excess fluid volume collected in minoxidil cycles was 1123.8 +/- 1119 mL versus 145.2 +/- 743.6 mL in the minoxidil-free cycles (p = 0.004). The mean creatinine clearance, urea nitrogen clearance, and protein losses were comparable in minoxidil cycles and the minoxidil-free cycles. Six patients developed hypotension during the minoxidil cycles, corrected by normal saline, but no other important side effects were noted. It is concluded that i.p. minoxidil selectively increases ultrafiltration without influencing solute clearance in peritoneal dialysis. PMID- 8728172 TI - Theoretical analysis of osmotic agents in peritoneal dialysis. What size is an ideal osmotic agent? AB - In this article the difference between osmotic fluid flow (ultrafiltration) as driven by osmotic pressure and diffusion through thin leaky membranes is discussed. It is pointed out that water transport induced by osmosis is fundamentally different from the process of water diffusion. Applying modern hydrodynamic pore theory, the molar solute concentration and the solute concentration in grams per 100 mL, exerting the same initial transmembrane osmotic pressure as a 1% glucose solution, was investigated as a function of solute molecular weight (MW). It was then assumed, base on experimental data, that the major pathway responsible for the peritoneal osmotic barrier characteristics is represented by pores of radius approximately 47 A. With increasing solute radius, the osmotic reflection coefficient (sigma) and, hence, the osmotic efficiency per mole of solute will increase. However, simultaneously, the molar concentration per unit solute weight will decrease. The balance point between these two events apparently occurs at a solute MW of approximately 1 kDa. An additional advantage of using solutes of high MW as osmotic agents during peritoneal dialysis (PD), rather than increased osmotic efficiency per se, lies in the fact that large solutes, due to their low peritoneal diffusion capacity, will maintain a sustained rate of ultrafiltration (osmosis) over a prolonged period. To illustrate this, we have performed computer simulations of peritoneal fluid transport according to the three-pore model of peritoneal permselectivity. According to these simulations, 4% of an 800 Da polymer solution (+50 mmol/L above isotonicity) will produce the same cumulative amount of intraperitoneal fluid volume ultrafiltered (UF) during 360-400 minutes as 4% of a 2 kDa polymer solution (+20 mmol/L) or 6.5% of a 10 kDa polymer solution (+6.5 mmol/L) having the same electrolyte concentration as dialysis solutions conventionally used for PD. Similar cumulative UF volumes (during 400 minutes) can be obtained by a 2.5% glycerol (+272 mmol/L) or a 3.2% glucose-containing dialysis solution (+177 mmol/L) with conventional electrolyte composition. PMID- 8728173 TI - Optimal use of glucose polymer (icodextrin) in peritoneal dialysis. AB - The osmotic effectiveness of glucose polymer is now well established. The relative inertness of this macromolecular compound has been the key factor in its success as the first "colloid" osmotic agent in clinical use. In its present form, it produces sustained ultrafiltration for up to 12 hours, and a daily overnight use would obviate the need for hypertonic exchanges, especially 3.86% glucose. In addition, it could be used in automated peritoneal dialysis regimes to enhance ultrafiltration and solute clearance during the daytime. Preliminary reports also indicate that it is beneficial in diabetic patients and in some patients who have lost ultrafiltration. The new "bimodal" formulations look promising, with the potential to replace all the currently used hyperosmolar exchanges with physiological solutions. Although systemic accumulation of glucose polymer breakdown products occurs, it reaches steady-state levels quickly (within two weeks) and remains stable throughout the duration of polymer use. In the long term study, these levels of maltose and oligosaccharides over three-and-a-half years represent the longest exposure of these substances in uremic patients without any clinical or metabolic adverse effects and provide an important evidence of safety. Future work based on studies that are ongoing suggest that a family of physiological solutions ("bimodal" preparations in iso-osmolar combination) could be available, and the individual's dialysis prescription could be tailored to take into account the ultrafiltration, metabolic needs, as well as the long-term viability of the membrane. Glucose polymer will be a key component of such solutions. PMID- 8728174 TI - Bicarbonate: the alternative buffer for peritoneal dialysis. AB - For a long time bicarbonate, the physiological buffer of the body, was suggested to be the best buffer for peritoneal dialysis. However, since the production of bicarbonate-containing solutions is associated with technical problems, lactate was favored. To avoid the well-known disadvantages of lactate solution concerning biocompatibility and possible metabolic side effects, different attempts have been made to use bicarbonate as a buffer in peritoneal dialysis. One of the major approaches was the total replacement of lactate by bicarbonate combined with storage of the fluid in a specially designed double-chamber bag. Further solutions of the above-mentioned problem were the on-line preparation of bicarbonate fluids for intermittent peritoneal dialysis, the addition of bicarbonate just before use, the combination of bicarbonate with organic acids, or its combination with the dipeptide glycylglycine as a stabilizing agent. By now, the beneficial effect of the neutral bicarbonate fluid, for example, on cell viability and cell functions, has been demonstrated in many different in vitro and animal studies. However, only few reports on clinical experience have been published. These investigations demonstrated independently that bicarbonate fluids diminish inflow pain, are well tolerated by the patients, and may correct metabolic acidosis of uremic patients. A controlled randomized multicenter trial using 34 mmol/L bicarbonate for at least three months confirmed that bicarbonate is as efficacious as lactate in equimolar concentrations. Concomitant investigations on energy metabolism and redox state of red blood cells and phospholipid secretion of mesothelial cells additionally demonstrated the improvement of cell function with bicarbonate solutions. For some patients with severe metabolic acidosis the bicarbonate concentration used in the multicenter trial seemed to be too low. Thus, a fluid containing a higher bicarbonate concentration was tested in a pilot study resulting in the expected significant increase of arterial bicarbonate levels. In summary, bicarbonate-containing peritoneal dialysis solutions are a promising alternative to lactate, especially if bicarbonate concentrations are adjusted individually to the patient's need. PMID- 8728176 TI - In vitro studies to assess biocompatibility of peritoneal dialysis solutions: strengths and limitations. PMID- 8728175 TI - Glucose degradation and cytotoxicity in PD fluids. PMID- 8728177 TI - Peritoneal host defense mechanisms. PMID- 8728178 TI - Acid-base balance with different CAPD solutions. AB - Our objective is to investigate transperitoneal buffer fluxes with solution containing lactate and bicarbonate, and to compare the final effect on body base balance of the two solutions. One hundred and four exchanges, using different dwell times, were performed in 52 stable continuous ambulatory peritoneal dialysis (CAPD) patients. Dialysate effluent lactate and bicarbonate and volumes were measured. Net dialytic base gain was calculated. Patients' acid-base status and plasma lactate were determined. In lactate-buffer CAPD solution, lactate concentration in dialysate effluent inversely correlated with length of dwell time, but did not correlate with plasma lactate concentration and net ultrafiltration. Bicarbonate concentration in dialysate effluent correlated with plasma bicarbonate and dwell time but not with ultrafiltration. The arithmetic sum of the lactate gain and bicarbonate loss yielded the net dialytic base gain. Ultrafiltration was the most important factor affecting net dialytic base gain. A previous study demonstrated that in patients using a bicarbonate-buffered solution the net bicarbonate gain is a function of dwell time, ultrafiltration, and plasma bicarbonate. By combining the predicted data of the dialytic base gain with the calculated metabolic acid production, an approximate body base balance could be obtained with both lactate- and bicarbonate-buffered CAPD solutions. The body base balance in CAPD patients is self-regulated by the feedback between plasma bicarbonate concentration and dialytic base gain. The level of plasma bicarbonate is determined by the dialytic base gain and the metabolic acid production. This can explain the large interpatient variability in acid-base correction. Bicarbonate-buffered CAPD solution is equal to lactate solution in correcting acid-base disorders of CAPD patients. PMID- 8728179 TI - Effect of different buffers on the biocompatibility of CAPD solutions. AB - Commercially available solutions for continuous ambulatory peritoneal dialysis (CAPD) affect the viability and function of the cells in the peritoneal cavity. The low biocompatibility of the solutions may be caused by a low pH, hyperosmolality, high glucose content, and lack of potassium, glutamine, and other components essential for normal cellular functions. The nature of the buffer employed is also important for the cytotoxicity of the solutions. Lactate, the most frequently used buffer, has been shown to inhibit cellular functions important for the peritoneal defense system including phagocytosis, bacterial killing, and secretion of cytokines. It is generally believed that the cytotoxicity of lactate is caused by lowering of intracellular pH and impairment of metabolism due to changed redox potentials. However, the cytotoxicity of lactate is highly dependent upon the pH of the solutions, indicating that passive or active diffusion across the cell membrane is determining the effects of lactate. Bicarbonate has been heavily advocated as an alternative buffer because it is the most important naturally occurring buffer in plasma and it enables a pH of approximately 7.4 in the solutions. However, due to sedimentation of calcium carbonate (CaCO3) and production of toxic glucose metabolites it is difficult to prepare and store bicarbonate-based solutions. Moreover, investigations have revealed that even bicarbonate-based solutions are not optimal regarding biocompatibility, presumably due to a paradoxical intracellular acidification caused by influx of carbon dioxide (CO2). More recently, the effect of other buffers such as pyruvate and histidine have been examined. Especially pyruvate is a promising new buffer candidate. PMID- 8728180 TI - Heat-sterilized PD fluid blocks leukocyte adhesion and increases flow velocity in rat peritoneal venules. AB - Data from cell culture experiments indicate that heat sterilization of peritoneal dialysis (PD) fluids produces cytotoxic glucose degradation products. The present vital microscopic study investigated the effects of different sterilization methods on the biocompatibility of PD fluids. Thus, heat-sterilized (commercially obtained and experimentally produced) and filter-sterilized PD fluids (pH = 5.30 5.40; 1.5% glucose) were compared with Tyrode buffer, with respect to the effects on microvascular blood flow velocity and leukocyte adhesion in the rat mesentery. Exteriorization of the mesentery produced a mild inflammation, known from the literature and characterized by the adhesive rolling of leukocytes along venular walls. Superfusion of the mesentery with filter-sterilized PD fluid had no significant effects on leukocyte rolling or flow velocity in venules 25-40 microns in diameter compared with buffer superfusion. Heat-sterilized PD fluid decreased the concentration of rolling leukocytes and increased flow velocity significantly, as compared with buffer and filter-sterilized PD fluid. The results indicate that heat sterilization of PD fluids produces substances that interact with microvascular tone and leukocyte-endothelial adhesion, which hypothetically could impair the acute, granulocyte-mediated defense against bacterial infections. PMID- 8728181 TI - Targets in CAPD and APD prescription. PMID- 8728182 TI - The impact of peritoneal permeability and residual renal function on PD prescription. AB - Our objective was to evaluate the impact of peritoneal transport characteristics and residual renal function on peritoneal clearance and to determine the dialysis volume required to achieve targets for KT/V and weekly creatinine clearance (WCC) in patients with differing weights, renal function, and transport status. Retrospective analysis and mathematical model simulation of urea and creatinine clearance were used. This review demonstrates the important contribution of both residual renal function and peritoneal transport in attaining target values for KT/V and WCC. The limitations of a standard 4 x 2 L dialysis prescription are highlighted in anuric patients and those with low peritoneal transport. In addition, the limitations of short dwell, rapid exchange dialysis modalities are emphasized, especially in patients with low and low average peritoneal transport where daily clearance is demonstrated to be considerably less than conventional continuous ambulatory peritoneal dialysis (CAPD). Attainment of proposed targets for KT/V (1.7) and WCC (50 L) is greatly dependent on residual renal function. In CAPD, peritoneal transport characteristics determine urea clearance through an effect on ultrafiltrate. Thus low transporters of identical weight will have greater values for peritoneal dialysis KT/V. Creatinine clearance is considerably influenced by transport status. The majority of low and low average groups will need some degree of renal function to achieve currently proposed targets. In these patients, conversion to high volume, short dwell modalities will further compromise small solute clearance unless daytime long dwells or tidal dialysis is instigated. PMID- 8728183 TI - Adequacy and quality of life with automated peritoneal dialysis. PMID- 8728184 TI - The predictive value of KT/V and peritoneal solute transport in CAPD patients is dependent on the type of comorbidity present. AB - Comorbidity, age, dialysis dose (KT/V(urea)), plasma albumin, and peritoneal function (D/P(creat) were measured cross-sectionally in 228 continuous ambulatory peritoneal dialysis (CAPD) patients, who were then followed up for a mean of two years. Comorbidity, utilizing a semi-quantitative score described previously, was the most powerful predictor of mortality in both univariate and multivariate analysis. Using univariate analysis, all the variables predicted outcome with statistical significance, mortality being associated with lower KT/V and plasma albumin and a higher D/P(creat). On multivariate analysis only comorbidity, age, and KT/V remained independent predictors. Data was further analyzed on the basis of type of comorbid condition. In those patients without comorbid disease (n = 127) neither KT/V, albumin nor D/P(creat) predicted outcome. In patients with clinical evidence of ischemic heart disease the KT/V was a significant predictor of favorable outcome. In those with clinical evidence of left ventricular function, mortality was significantly and independently associated with low plasma albumin, high D/P(creat), and KT/V. It is suggested that the concept of treatment adequacy in CAPD patients must include both measures of dialysis dose and peritoneal function, particularly in the context of the patient's comorbidity. PMID- 8728185 TI - Survival of CAPD patients in a center using three two-liter exchanges as standard regime. AB - Our objectives were to study long-term patient survival and dialysis adequacy of continuous ambulatory peritoneal dialysis (CAPD) patients treated with a standard regime of three 2-L daily exchanges. It was a retrospective analysis of patient survival and cross-sectional analysis of adequacy of dialysis. The setting was a dialysis unit in a tertiary referral center of a teaching hospital. All patients (n = 507) accepted into the CAPD program from 1983 to June 1994, were analyzed for survival. Adequacy of dialysis was analyzed in all existing patients in 1993 to 1994. The overall patient survival was 93%, 71%, and 57% at one, three, and five years, respectively. The three-year survival rate was 40% for diabetics and 78% for nondiabetics. It was 86%, 85%, 64%, and 43% for patients aged < 35, 35 50, 50-65, and over 65 years, respectively. The mean weekly KT/V of 201 existing CAPD patients was 1.76, and creatinine clearance was 57 L/week/1.73 m2. KT/V by dialysis was only 1.57. The age- and disease-adjusted survival in our center was comparable to centers that used standard four 2-L exchange regimes, despite a substantially lower mean KT/V and creatinine clearance (CrCl). PMID- 8728186 TI - How to reach optimal creatinine clearances in automated peritoneal dialysis. AB - This paper summarizes the basis of prescription for automated peritoneal dialysis (APD) established during a French national conference on APD. Clinical results and literature data show that peritoneal clearances are closely determined by peritoneal permeability and hourly dialysate flow rate, independently of dwell time or number of cycles. With APD, peritoneal creatinine clearance increases according to the hourly dialysate flow rate to a maximum (plateau), then decreases because of the multiplication of the drain-fill times. The hourly dialysate flow giving the maximum peritoneal creatinine clearance is defined as the "maximal effective dialysate flow" (MEDF). MEDF is higher for high peritoneal permeabilities: MEDF is 1.8 and 4.2 L/hr with nocturnal tidal peritoneal dialysis (TPD) for a 4-hr creatinine dialysate-to-plasma ratio (D/P) of 0.50 and 0.80, respectively. With nightly intermittent peritoneal dialysis (NIPD), MEDF is 1.6 and 2.3 L/hr for a D/P of 0.50 and 0.78, respectively. Under these conditions, tidal modalities can only be considered as a way to increase the MEDF. Using the MEDF concept for an identical APD session duration, the maximal weekly normalized peritoneal creatinine clearance can vary by 340% when 4-hr D/P varies from 0.41 to 0.78. APD is not recommended when 4-hr creatinine D/P is lower than 0.50. However, the limits of this technique may be reached at higher peritoneal permeabilities in anurics because of the duration of sessions and/or the additional exchanges required by these patients. PMID- 8728187 TI - How much extra does "adequate" peritoneal dialysis cost? AB - Our objective was to investigate the extra cost associated with implementing an adequacy program in peritoneal dialysis (PD) and to evaluate the cost effectiveness of a variety of PD prescriptions. This was a cross-sectional study of all 37 patients attending the PD clinic at a university teaching hospital. Extra costs incurred on dialysate, tubing, and cyclers as a consequence of implementing an adequacy program in PD were measured. Costs per unit KT/V for a variety of PD prescriptions were also calculated. Thirteen patients (35%) required an alteration in prescription for adequacy reasons. The average extra costs incurred for all patients was Cdn. $2,323 per annum, which represents a 16% increase per patient. The most cost-effective prescriptions were high volume continuous ambulatory peritoneal dialysis (CAPD) and automated peritoneal dialysis (APD) with two daytime dwells. The least cost-effective prescriptions were day dry APD and high volume APD with only one daytime dwell. Significant extra expense is incurred when an adequacy program is implemented in PD. This is mainly due to the requirement to put more patients on APD. An alternative pricing policy for PD is suggested. PMID- 8728188 TI - Compliance study in peritoneal dialysis using PD Adequest software. AB - Poor compliance in peritoneal dialysis (PD) is a significant cause of dropout and morbidity. PD Adequest software, which, through a mathematical model, predicts the effect of the dialysis prescription on the basis of the peritoneal transport, may be used to identify the noncompliant patient. Fifty patients from two dialysis centers, aged 65.9 +/- 1.5 years and on PD for 28.6 +/- 4.7 months, were studied. A peritoneal equilibration test (PET) was carried out and 24-hour urine and dialysate were collected. Total weekly creatinine clearance (CrCl, L/week/1.73 m2) was calculated, as well as the glomerular filtration rate [(GFR), mL/min, mean CrCl and urea nitrogen clearance (UNCI)]. The dialytic schedules used were then introduced into the program and the parameters were recalculated using the software model. Nine patients considered noncompliant from their case histories were used to assess the differences of reference between expected and measured values. The control group was significantly different from the noncompliant group in the percentage of the CrCl and the serum creatinine (sCR) differences. The noncompliance threshold value was calculated from the mean of the lower 95% confidence interval of the compliant group and the higher one of the noncompliant group (-5.3%) for CrCl and vice versa for sCR (+10%), which behaved to the contrary. Reassessing the patients, 11 (22%) were identified as probably noncompliant. PMID- 8728189 TI - Appetite in CAPD patients. PMID- 8728190 TI - Normalization of protein equivalent of nitrogen appearance and dialytic adequacy in CAPD. AB - Caloric-proteic malnutrition is frequently encountered in peritoneal dialysis and is associated with an increased risk of morbidity and mortality. Our paper aims to assess any greater reliability of protein equivalent of nitrogen appearance (PNA) normalization to desirable body weight (dBW) compared to actual body weight (aBW) and resulting implications for the relationship between dialytic adequacy and protein intake in continuous ambulatory peritoneal dialysis (CAPD). We studied 36 patients on CAPD, 24 male and 12 female (aged 66.6 +/- 10.2 years, 24 +/- 29 months on dialysis), collecting dialysate and urine over 24 hours (126 samples) to calculate the PNA according to Randerson and the total weekly KT/V. The total body muscle mass (TBMM) was calculated by anthropometry and the dBW according to Metropolitan Life Insurance tables. Finally, PNA was normalized to aBW (aPNA, g/kg/day) and to dBW (dPNA, g/kg/day). Average aBW proved to be higher than dBW (66.0 +/- 11.1 vs 59.8 +/- 6.9 kg, p < 0.0001) and aPNA lower than dPNA (0.96 +/- 0.31 vs 1.08 +/- 0.3 g/kg/day, p < 0.005). Compared to aPNA, dPNA correlates better with both blood urea nitrogen (BUN) (R2 = 0.702 vs 0.614) and KT/V (R2 = 0.348 vs 0.306). The TBMM is higher in the group with dPNA > or = 1.0 vs < 1.0 g/kg/day (25.5 +/- 0.6 vs 23.1 +/- 0.7 kg, p < 0.02) while, paradoxically, it is lower in patients with aPNA > or = 1.0 vs < 1.0 g/kg/day (22.8 +/- 0.8 vs 25.4 +/- 0.6 kg, p < 0.01). The KT/V of the patients with dPNA < 0.8, 0.8-1.2 and > 1.2 g/kg/day proved to be different (1.52 +/- 0.06 vs 1.80 +/- 0.03 vs 2.04 +/- 0.04, p < 0.005). On analysis of the linear regression, dPNA = 1.0 and 1.2 g/kg/day corresponds to KT/V values of 1.7 and 2.05, respectively. We consider dPNA to be more suitable then aPNA for the correct assessment of protein intake, and a weekly KT/V of 1.7-2.05 as being sufficient to guarantee satisfactory dPNA. PMID- 8728191 TI - Predictors of survival in continuous ambulatory peritoneal dialysis patients: a five-year prospective study. AB - Our objective was to examine the influence of various demographic, clinical, and enrollment biochemical variables on the long-term survival of continuous ambulatory peritoneal dialysis (CAPD) patients. This was a prospective cohort study investigating the relationship between demographics and enrollment biochemical markers and mortality in CAPD patients in a CAPD unit in a large tertiary care teaching hospital. One hundred and sixty-nine patients in the CAPD program were enrolled between 1989 and 1994, and were followed up to 60 months. Independent predictors of mortality determined by Cox proportional hazards model included age, diabetes, serum albumin and creatinine. Enrollment level of serum albumin, and creatine can predict mortality in CAPD patients up to 60 months. Markers of visceral and somatic nutrition at enrollment are important predictors of mortality in CAPD patients up to five years. PMID- 8728192 TI - Nutritional status of CAPD patients at three years. AB - A nutritional assessment was carried out in 63 patients starting treatment from April 1990 up to December 1993. Anthropometric measurements were performed showing a prevalence of protein-calorie malnutrition (PCM) of 21% in a total of 142 clinical surveys carried out in the above-mentioned sample. A steady state of albumin levels in plasma was verified during a three-year follow-up period at a lower level than that of the control group. The patients' lymphocytic profile throughout the study was characterized by lymphopenia and decreased B and T8 lymphocytes. During the first two years of continuous ambulatory peritoneal dialysis (CAPD), a high percentage of patients met the "adequacy" dialysis criteria as residual renal function plays an important role as regards treatment. PMID- 8728193 TI - Impact of the liver on peritoneal transport. AB - Previously, we developed a distributed model of plasma-peritoneal small solute diffusion for specific tissues surrounding the peritoneal cavity and related the transport coefficients to the mass transport coefficient (MTC) of the "peritoneal membrane" model. Based on this theoretical analysis, we calculated tissue specific MTCs for sucrose from microvascular data in the literature and found that the MTC for the liver was five times the magnitude of other tissues. We hypothesized that the liver was potentially the most significant single transport organ during peritoneal dialysis. To test this hypothesis, we measured the mass transfer from the plasma to fluid contained in diffusion chambers, which were glued to one of four tissues surrounding the peritoneal cavity. We determined that the rate of small solute transport from the plasma to each diffusion chamber was similar for all four tissues. We calculated the MTC of the liver to be no greater than other visceral or parietal surfaces. We therefore disproved our hypothesis concerning the liver. We conclude that the importance of a particular tissue to plasma-peritoneal transport is primarily dependent on the surface are exposed to the dialysis solution. PMID- 8728194 TI - The role of the liver in the pathogenesis of hyperlipidemia in patients with end stage renal disease treated with continuous ambulatory peritoneal dialysis. PMID- 8728195 TI - The impact of intraperitoneal glucose and insulin on the liver. PMID- 8728196 TI - Peritoneal dialysis in liver disorders. AB - The purposes of this paper is to review the specific role of peritoneal dialysis (PD) in patients with liver disorders. We will pay attention to the confluence of liver diseases and situations for which chronic dialysis treatment is required. Hemodialysis (HD) and peritoneal membranes are safe barriers against the passage of the hepatitis C virus; consequently, while peritoneal effluent or HD ultrafiltrate drained from hepatitis B patients/carriers is infective, that from hepatitis C patients does not appear to present this risk. An important issue is horizontal transmission, which appears to occur with both viruses in HD units, and which is absent in peritoneal dialysis units. The incidence of hepatitis C among continuous ambulatory peritoneal dialysis (CAPD) patients is quite low, while it may reach almost 50%-60% of HD patients in some units. While hepatitis C transmission mechanisms are not completely understood and a vaccine is not available, PD provides some degree of protection when compared with HD, for and stage renal disease patients. In summary, our experience and that of others, with a total of 19 PD-treated chronic liver disease patients, supports CAPD as the treatment of choice for cirrhotic patients with ascites who require chronic dialysis. Data on peritoneal diffusion of low molecular weight substances revealed a marked increase in most patients. The ultrafiltration capacity was clearly augmented with respect to noncirrhotic patients, making the use of hypertonic bags unnecessary. Hemodynamic tolerance was excellent. Complications and death were mainly related to liver disease complications. Spontaneous bacterial peritonitis (SBP), caused by gram-negative germs, is the most important complication directly related to ascites and may have some points in common with PD-related peritonitis. However, and in contrast to most PD peritonitis, two pathogenetic mechanisms have been suggested for SBP: (1) translocation of bacteria from the gut to the mesenteric lymph nodes, and (2) bacteremia in these patients is secondary to the general abnormal host defense mechanisms. Local factors such as intrahepatic shunting and the impairment of bactericidal activity in ascitic fluid favor the bacteria ascites. The hypothesis of a direct transmural contamination from bowel to ascitic fluid has been relegated to secondary bacterial peritonitis. Would cirrhotic patients with temporal or permanent renal function compromise benefit from peritoneal catheter placement and other PD practices to perform repetitive small ascitic drainages at home? Perhaps the time has arrived when hepatologists and PD nephrologists begin to work shoulder to shoulder in this particular field, as we have a common problem, the peritoneal cavity filled with fluid. PMID- 8728197 TI - Is atherosclerosis accelerated by CAPD? AB - Cardiovascular complications in renal replacement therapy remain prevalent today. The question, "Is atherosclerosis accelerated in peritoneal dialysis (PD) patients," has not been resolved. Many cross-sectional studies have revealed that there are more atherogenic lipid profiles in continuous ambulatory peritoneal dialysis (CAPD) than in other dialytic modalities. However, it is not certain that CAPD per se may contribute to lipid abnormalities in continuing PD for a long time. Therefore, we tried to assess whether CAPD itself may change lipid profiles in the long-term period on CAPD. We measured conventional lipid profiles in 16 stable CAPD patients in whom total cholesterol (T.chol.) levels remained under 240 mg/dL at the time of starting observation. Diabetic end-stage renal disease (DMESRD) patients were excluded from this study. Blood sampling was performed under strict conditions indicating overnight fasting with 10 hours dwell of 2 L of 1.5% Dianeal. Plasma levels of T.chol., triglyceride, high density lipoprotein (HDL), low-density lipoprotein (LDL), and atherogenic lipoprotein (apo B/A-I) did not show significant changes by serial measurements from the sixth month to the thirtieth month following the commencement of CAPD as their initial dialysis treatment. Recently it is reported that high plasma levels of Lp(a) lipoprotein are an independent risk factor in cardiovascular events in renal replacement therapy because Lp(a) has a close relation to intravascular thrombosis and acceleration of atherosclerosis. We found that the incident of vascular accidents was five episodes among 33 patients with higher (> 31 mg/dL) Lp(a) levels, while there was only one episode in 45 patients with lower (< 30 mg/dL) Lp(a) levels (p < 0.05, odds ratio: 7.9). However, severity of aortic calcification and incidence of positive treadmill test showed no difference in these two groups. No significant correlation was observed between duration on CAPD and plasma Lp(a) levels. From the fact mentioned above, we speculate that CAPD does not essentially affect lipid profiles if determined under the strict condition of blood sampling. In order to evaluate the atherosclerosis noninvasively, we have measured aortic pulse wave velocity (AoPWV) in 33 stable CAPD patients excluding those with DMESRD by two years' interval. Fifteen cases (46%) increased in AoPWV, however, lipid profiles did not differ from those of nonadvanced patients. Calcification of arteries is further evidence of acceleration in atherosclerotic change. Therefore, we have graded severity of abdominal aortic calcification into three categories: grade I denoted nil calcification, grade II denoted patchy calcification, and grade III denoted calcification along the entire abdominal wall, having a lead-pipe shape by lateral view of plain abdominal x-ray film. There were significant differences in the duration of CAPD (grade I: 41 months, grade II: 60 months, grade III: 68 months). AoPWV showed least in the grade I group, faster in grade II, and fastest in grade III, while lipid profiles did not show significant differences in three categories. From the analysis of serial changes of lipid profiles, AoPWV and aortic calcification, CAPD may present some risk of accelerating atherosclerosis, at least in some patients on long-term treatment. Risk factors contributing to acceleration of atherosclerosis is result not from lipid abnormalities, but from other factors which remained to be seen, for example, abnormalities in calcium metabolism. PMID- 8728198 TI - PD treatment for severe congestive heart failure. AB - Our objective was to evaluate if peritoneal dialysis (PD) could improve survival of patients with progressive severe congestive heart failure resistant to drug therapy. The patients were selected by the cardiologist in cooperation with a nephrologist, including patients not responding to conventional medication with an expected fatal outcome within the next months. The study included 16 consecutive patients with a chronic progressive severe refractory heart failure (sHF) of NYHA class III (n = 6) or IV (n = 10) who did not respond to diuretics and angiotension converting enzyme (ACE) inhibitors. They had a mean age of 60 years (+/- 14, range 30-75, median 62 years). Nine of the patients had sHF as the only reason for initiating PD (all NYHA IV), while 7 also needed dialysis due to uremia. Five of 7 had been on hemodialysis but switched to PD due to a progressive congestive sHF. In 2 patients, PD was decided already at start of dialysis therapy due to the severity of their heart failure. The reason for sHF was: valvular dysfunction (n = 5) with defect prosthesis (n = 3); in the course of a myocardial infarction (n = 4); and cardiomyopathy (n = 4). Tenckhoff catheters were inserted under local anesthesia and ultrafiltration was started and maintained until discharge. The survival time and change in heart size by x ray was used for analyses. All patients improved their stage of congestive heart failure by NYHA classification already during the first month. Six patients died during the follow-up period due to cardiac reasons (sudden death, relapse of sHF) after a mean of 10.7 months (+/- 3.7, range 1-24 months). Ten were alive after a median observation period of 10 months (+/- 12.5, range 1-36 months). Heart size was reduced in 15 of the patients. Three of the patients with sHF but without uremia could stop the PD. The results showed that ultrafiltration by PD was easy to perform despite low initial blood pressure. The sHF was reduced and life span was prolonged with improved quality of life. PMID- 8728199 TI - Lipoprotein (a) levels in patients undergoing continuous ambulatory peritoneal dialysis. AB - Most researchers have found increases of lipoprotein (a) [Lp(a)] in uremic patients, as well as in those undergoing hemodialysis (HD) or continuous ambulatory peritoneal dialysis (CAPD). The mechanisms for this increase remain unclear. We studied 71 patients undergoing CAPD, 48 men and 23 women. According to the time spent on CAPD, the patients were divided into three groups: group 0: 29 patients at the starting off point of dialysis treatment; group I: 22 patients with an average stay of 15.2 months; group II: 20 patients with an average stay of 69.3 months on CAPD. We have only observed significant increases of Lp(a) levels in those patients initiating the dialysis, but no significant differences are found in the other groups undergoing CAPD for longer periods when compared to the control group. We found no significant relation between Lp(a) levels and peritoneal protein loss, and not with absorption of glucose from the dialysate either. We have found a positive and significant correlation between Lp(a) levels and urinary protein loss (r = 0.41; p < 0.001). It is possible that an element associated with proteinuria might have an effect on the metabolism of Lp(a) in CAPD patients. PMID- 8728200 TI - Impact of nutritional status on serum lipoprotein (a) concentration in patients undergoing continuous ambulatory peritoneal dialysis. PMID- 8728201 TI - Increased serum level of total homocysteine in CAPD patients despite fish oil therapy. AB - It has been shown that serum total homocysteine (HC) is a risk factor for vascular disease which characterizes endothelial damage. The incidence of vascular disease is increased in continuous ambulatory peritoneal dialysis (CAPD) patients. Our aim was to investigate: (1) whether concentration of HC correlates with atherosclerotic and inflammatory events, and (2) if fish oil therapy can retard the disturbance in lipid metabolism which promotes atherosclerosis. Fourteen patients with various degrees of impaired peritoneal clearance and lipid metabolism were observed. In all patients the serum HC was elevated. Seven patients were treated with fish oil for three months. The results indicate an average increase of HC (+18%), total cholesterol (+6.6%), aggregation of erythrocytes (+9%), and an average decrease of dialysate-to-plasma creatinine (D/P) ratio (-7%), deformability of erythrocytes (-8%), and normalization of elevated soluble interleukin-2 receptor (sIL-2R) values. Regression analysis of all data demonstrated a significant correlation between HC and parameters of lipid metabolism and hemorheology. There were no significant correlations between HC and peritoneal function and serum cytokine levels. We conclude that the treatment in CAPD patients with fish oil did not improve the lipid metabolism disturbances in atherosclerosis and peritoneal function. Elevated HC confirms the progression of the disease. PMID- 8728202 TI - Eicosanoid precursors: potential factors for atherogenesis in diabetic CAPD patients? AB - Prostaglandins, thromboxanes, and other eicosanoids represent a widespread lipid mediator system for intercellular signalling, and, hence, have multiple cellular actions. Thus it is not surprising that numerous events in the pathogenesis of atherosclerosis are associated with an altered formation of eicosanoids. To reconsider the availability of eiconsanoid precursors as one possible cause of atherogenesis, the dietary intake and the serum concentrations of arachidonic acid (AA) and eicosapentaenoic acid (EPA) were determined in patients with high risk for atherosclerosis on continuous ambulatory peritoneal dialysis (CAPD) with and without diabetes in comparison to healthy controls and diabetic patients without late complications. The factor AA/EPA in serum was created as a marker for the atherosclerosis risk. The setting was in a CAPD unit in one city hospital. There were 26 CAPD patients [9 with insulin-dependent diabetes mellitus (IDDM), 9 with noninsulin-dependent diabetes mellitus (NIDDM), and 8 without diabetes], 27 IDDM without late complications, and 41 healthy control persons. The AA levels in serum were significantly higher in all of the CAPD groups. In contrast, the EPA concentrations in serum were significantly lower in the CAPD groups, with the lowest EPA levels found in the CAPD-IDDM group. Therefore, the factors AA/EPA in serum were significantly higher in all of the CAPD groups, and again significantly higher in the CAPD-IDDM group than in the other CAPD groups. No differences in the amount of dietary intake of AA existed between the groups. The daily intake of EPA was significantly highest in the control group. Higher concentrations of AA and a lack of n-3 fatty acids lead in the presence of a reduced prostaglandin I2 biosynthesis, to a higher formation rate of potentially proatherogenic metabolites such as thromboxane A2, a vasoconstricting and platelet aggregating agent. Thus, the quotient AA/EPA could possibly be used as a marker of atherogenicity in the future. PMID- 8728203 TI - Peritoneal dialysis and endocrinology. PMID- 8728204 TI - Endocrinology and PD: optimal treatment of anemia in peritoneal dialysis patients. PMID- 8728205 TI - Dialysis in patients with diabetic nephropathy: CAPD versus hemodialysis. AB - Diabetic nephropathy has emerged as a major cause of ESRD over the past decade, being the most prevalent cause of ESRD requiring dialysis in North America (United States and Canada) and the second highest in the incidence rate in Europe, Japan, Korea, Australia, and New Zealand. A greater proportion of older patients and of patients with diabetic nephropathy and other comorbid conditions has been treated with CAPD. Despite the preferential use of CAPD to treat a high risk group of patients, the overall and/or selection-adjusted mortality was similar between HD and CAPD groups. Among diabetic patients, selection-adjusted mortality was similar between HD and CAPD or lower in CAPD than in HD, the difference being greatest among younger patients and significant through the age of 52, or higher in CAPD than in HD with higher risk of death for older diabetics (age > or = 50 years), but with similar risk among younger diabetics (age < 50 years). Technique survival was also variably reported as similar between HD and CAPD, lower, or higher with CAPD compared to HD. Diabetic CAPD patients had more hospital admissions and more days in the hospital and higher withdrawal rates from dialysis compared to diabetic HD patients. These disparate results of patient and technique survival between HD and CAPD in diabetic patients may have resulted from patient selection criteria with different comorbid conditions on entrance to dialysis, quantity of dialysis, and other unrecognized factors. Prospective randomized studies are needed to assign a cause-and-effect relationship between the choice of dialysis modality and patient and technique survival among patients with diabetes mellitus as well as with all other diagnostic categories. PMID- 8728206 TI - Peritoneal dialysis in diabetics. Optimal insulin therapy on CAPD: intraperitoneal versus subcutaneous treatment. PMID- 8728207 TI - Management of foot problems in diabetes. AB - Many diabetic foot complications are preventable. This requires a team approach, aiming to identify the high-risk patient and provide appropriate education and foot care. An established ulcer needs careful management with the emphasis on pressure relief and establishing a good blood supply. PMID- 8728208 TI - Survival of diabetic patients on peritoneal dialysis or hemodialysis. AB - Our objective was to analyze the survival of diabetic patients on renal replacement therapy and to compare their survival on extracorporeal and on peritoneal dialysis. All data regarding diabetic patients admitted to dialysis between 1 January 1983 and 31 December 1993 were collected by means of individual patient questionnaires sent to all of the 44 regional Renal Units (100% answers) of Lombardy, Italy. Cox proportional hazards model, stepwise procedure, was applied in order to select the covariates significantly associated with survival. Age (at baseline), sex, type of diabetes, initial modality of treatment (hemodialysis or peritoneal dialysis), and initial clinical risk factors (malignancies, serious heart disease, vascular disease, cirrhosis of the liver, cachexia) were considered. Descriptive analysis of survival was performed using the Kaplan-Meier technique. The survival of all diabetic patients (895) was 86.5% at one year, 52% at three years, and 34% at five years. The main causes of the 488 deaths of diabetic patients were cardiovascular diseases (56%), cachexia (18%), and infections (11%). The relative death risk of patients on peritoneal dialysis versus those on hemodialysis, after taking into account the main comorbid conditions, did not significantly differ from 1, as estimated by the Cox proportional hazards regression model. Five-year survival of diabetic patients was 34%, and no differences were found between peritoneal dialysis and hemodialysis as far as mortality is concerned. PMID- 8728209 TI - Subcutaneous and intraperitoneal insulin therapy in diabetic patients on CAPD. AB - We evaluated in a cross-over manner the consequences of subcutaneously and intraperitoneally given insulin on glucose control, insulin sensitivity, and serum lipids in 8 type I diabetic patients on continuous ambulatory peritoneal dialysis (CAPD). The patients were treated with both subcutaneous and intraperitoneal insulin for at least three months. After each period, metabolic studies were performed. Despite significantly improved glycemic control (Hb A1c 10.00 +/- 0.38% after subcutaneous and 8.40 +/- 0.36% after intraperitoneal insulin, p = 0.01), serum lipids showed unfavorable changes. High-density lipoprotein (HDL)-cholesterol was significantly lower (1.28 +/- 0.18 mmol/L vs 0.88 +/- 0.06 mmol/L, p = 0.03) and low-density lipoprotein (LDL)/HDL-cholesterol ratio was higher (p = 0.025) during intraperitoneal insulin. Total cholesterol, LDL-cholesterol, and triglycerides were higher during intraperitoneal insulin administration. Severe hypoglycemic episodes were more common during subcutaneous than intraperitoneal insulin. It is concluded that, although intraperitoneal insulin administration offers significantly better glycemic control and insulin sensitivity than subcutaneous insulin, the effect of serum lipids is more disadvantageous possibly via a direct effect of insulin on the liver. PMID- 8728210 TI - Low turnover bone disease. PMID- 8728211 TI - Prescription of calcium concentration and PTH control. AB - The use of calcium-containing oral phosphate binders, introduced in an effort to avoid aluminum-containing compounds, had led to more frequent episodes of hypercalcemia. This prompted the introduction of continuous ambulatory peritoneal dialysis (CAPD) solutions with diminished calcium content. The problems raised by such solutions included stimulation of parathyroid hormone (PTH) secretion and long-term maintenance of calcium balance. Some of these issues can today be answered on the basis of controlled prospective trials. Variability of the rate of intestinal calcium uptake of bone turnover, of baseline parathyroid activity, and other factors make it necessary to individualize the indication for the use of CAPD solutions with low calcium content. PMID- 8728212 TI - Does calcitriol therapy improve muscle function in uremic patients. AB - The purpose of the present study is to investigate whether calcitriol therapy in uremic patients undergoing dialysis treatment can improve muscle function. In 8 uremic patients [2 on continuous ambulatory peritoneal dialysis (CAPD), 6 on hemodialysis (HD) treatment], calcitriol (Calcitriol, Roche) was given in the dose of 1 microgram/day for 15 months. At the beginning of therapy and every month, the following parameters in serum were determined: creatinine (Cr), calcium (Ca), phosphate (P), and parathyroid hormone (PTH). At the beginning and at the end of treatment, quantitative electromyography (EMG) was performed. We observed a slight increase of serum Cr during 15 months of treatment in 4 patients. Serum Ca, P, and PTH did not change significantly. The EMG revealed abnormal polyphasic motor nerve unit potentials of brief durations, a decrease in the amplitude, and fibrillation potentials. The EMG findings did not change significantly after calcitriol therapy, but all patients on physical examinations exhibited the disappearance of clinical manifestations of uremic myopathy. In conclusion, our findings suggest that vitamin D deficiency is one of the causes of uremic myopathy and a careful treatment with calcitriol can diminish muscle weakness in uremic patients. PMID- 8728213 TI - Serum procollagen type I carboxyterminal propeptide in CAPD and hemodialysis patients. PMID- 8728214 TI - A comparison of bone scans in uremic patients treated with intermittent peritoneal dialysis or hemodialysis. AB - The aim of the study was a comparative analysis of bone scans in uremic patients treated with intermittent peritoneal dialysis (IPD) or hemodialysis (HD). Bone scintigraphy was performed using technetium Tc 99m etidronate (EHDP) in 28 uremics (age 46.0 +/- 13.5 years, x +/- SD) on IPD for 3.1 +/- 3.0 months and 28 uremics (age 43.5 +/- 11.6 years) on HD for 47.3 +/- 33.9 months. Serum c terminal parathormone (cPTH) exceeded 5.3 +/- 3.3 and 6.8 +/- 3.5 times the upper normal limit of 1.4 ng/mL in IPD and HD patients, respectively. Despite significant differences in dialysis treatment duration in IPD and HD patients, an increased Tc 99m EHDP uptake in bones was shown with similar frequency, when all the groups were compared. However, in the group of patients with serum cPTH exceeding four times the upper normal limit (n = 30) or in the age group less than 45 years old (n = 26), a greater marker uptake was observed in HD patients. Significant differences (p < 0.05) were shown in the cranial vault: 33% of HD patients (n = 18) with higher cPTH and 47% of those less than 45 years old (n = 15) revealed an increased marker uptake, whereas it was not observed in any IPD patient. When scans of HD patients dialyzed less than (n = 11) and more than (n = 17) 30 months were compared, a significantly higher appearance of increased marker uptake was shown in cranial vault (41% vs 0%, p < 0.02) and in sacral bone (82% vs 36%, p < 0.02) in patients with longer dialysis. The latter group of HD patients also showed an increased marker uptake in cranial vault compared to the entire group of PD patients (41% vs 7%, p < 0.01). Our studies suggest that bone scan changes, indicating secondary hyperparathyroidism, progress significantly with prolongation of dialysis treatment, especially in patients with higher cPTH levels of younger age. PMID- 8728215 TI - Implantation techniques for peritoneal dialysis catheters. PMID- 8728216 TI - The peritoneal access. PMID- 8728217 TI - Peritoneal dialysis access technology: the Austin Diagnostic Clinic experience. PMID- 8728218 TI - Peritoneoscopic placement of Swan neck peritoneal dialysis catheters. AB - Peritoneoscopic placement of peritoneal dialysis catheters, although accomplished in only about 10% of dialysis centers, is a nonsurgical technique that fulfills requirements for safety and dependability. Over a 40-month period, 136 catheters were placed with the peritoneoscope, 135 of which were double-cuffed, Swan neck curled catheters, with a uniform radiopaque stripe. Patients were followed longitudinally for outcome. Catheters were placed in 44 diabetic patients, 1 human immunodeficiency virus (HIV)-positive patient, and 18 morbidly obese patients. No complications occurred as a direct result of placement. Catheters were used, on average, nine days after placement (many on days 1 to 4) usually with 1.5 to 2 L exchanges. With 1183 patient-months' experience, complications were few: 28 patients experienced catheter-related infections, and there were five leaks that resolved with supine, low-volume dialysis for several days. Leakage did not correlate with time of usage after placement. Of ten outflow/mechanical problems that required catheter removal, nine involved catheter migration, probably due to lack of attention during placement to orientation of the radiopaque stripe. One was due to a preperitoneal placement early in this institution's experience with the peritoneoscope. Five of the migrated catheters were removed and then successfully replaced with the peritoneoscope at the same sitting. Four patients requested surgical removal and replacement. Sixteen catheters were removed because of catheter-related infections: five refractory Staphylococcus aureus, six Pseudomonas aeruginosa, two fungal, two Serratia species, and one Mycobacterium chelonei. Actuarial life table analysis showed that at the end of the 40-month follow-up, 62% of the catheters were expected to survive. Because more than 50% survived, median catheter survival could not be calculated. The adverse responses were removal because of infection or catheter migration. Peritoneal dialysis catheter implantation with the peritoneoscope represents a safe and dependable method for catheter placement. Literature review and comparison indicate that catheter related complications are fewer and catheter longevity is better with peritoneoscopic placement than with surgical placement. Our experience with prompt postplacement utilization suggests the need for further evaluation of catheter break-in procedure with the peritoneoscope. PMID- 8728219 TI - A prospective randomized comparison of the Swan neck, coiled, and straight Tenckhoff catheters in patients on CAPD. AB - Our objective was to study the impact of peritoneal catheter configuration on continuous ambulatory peritoneal dialysis (CAPD)-related infections, mechanical complications, and patient dropout in a prospective randomized trial. Forty consecutive patients who were commencing CAPD were randomized to receive either a double-cuff, Swan neck coiled catheter or a double-cuff, straight Tenckhoff catheter, implanted by surgical technique. There was no significant difference in the peritonitis rate between the two groups. There was a lower rate of exit-site infection in the Swan neck group compared to the straight catheter group (0.29 vs 0.60 episodes/patient-year, p < 0.05). Catheter-tip migration occurred in 3 patients with the straight catheters compared to one patient with the Swan neck catheter. No patient had to discontinue CAPD because of mechanical complications. The number of CAPD patient dropouts was not significantly different between the two groups. The Swan neck configuration resulted in a significant reduction in the rate of exit-site infections. The coiled component of the catheter may lead to fewer episodes of catheter-tip migration. However, catheter configuration did not influence the number of technique failures. PMID- 8728220 TI - Exit-site care. PMID- 8728221 TI - Significance, epidemiology, and prevention of peritoneal dialysis catheter infections. AB - Catheter infections are a major cause of morbidity, catheter loss, and transfer to hemodialysis. These infections are mainly due to S. aureus. To date, treatment is less than optimal. Therefore, the primary goal should be prevention of catheter infections. Prevention is based on improving catheter design and implantation technique while using careful exit-site care. Prophylaxis with antimicrobials such as intranasal mupirocin or the use of silver-impregnated catheters appears promising as a way to reduce the risk of developing S. aureus infections. To optimize patient outcome, one must focus on these preventive measures. PMID- 8728222 TI - Long-term consequences of peritonitis. PMID- 8728223 TI - Prevention of infection dropout in PD. PMID- 8728224 TI - Impaired initial cell reaction in CAPD-related peritonitis. AB - Our objective was to determine the incidence of peritonitis episodes with an impaired initial cell reaction (IICR:neutrophil number < 100 x 10(6)/L) over a period of ten years, and to find possible explanations for this unusual presentation of peritonitis. A retrospective review of the files of continuous ambulatory peritoneal dialysis (CAPD) patients included in the CAPD program 1984 and 1993 was done. Analysis of cytokine and prostanoid patterns during four peritonitis episodes with an IICR was compared to 12 episodes with a normal initial cell reaction (NICR). Dialysate cell numbers and immunoeffector characteristics of peritoneal cells were compared in 7 IICR patients in a stable situation and a control group of 70 stable CAPD patients. The setting was a CAPD unit in the Academic Medical Center in Amsterdam. Thirty-five CAPD patients who had one or more peritonitis episodes with an IICR and a control group of 249 CAPD patients were included in the study. The incidence of peritonitis with an IICR was 6%. These episodes occurred more than once in 51% of the patients who presented with IICR. In 72% the cell reaction was only delayed: a cell number exceeding 100 x 10(6)/L was reached later. Staphylococcus aureus was significantly more frequently the causative microorganism compared to all peritonitis episodes (PE) that occurred during the study period. Patients with IICR had lower dialysate cell counts in a stable situation, compared to a control group (p < 0.01). This was caused by a lower number of macrophages and CD4 positive lymphocytes. The phagocytosis capacity of the macrophages appeared to be normal. In a comparison of four PE with an IICR and 12 episodes with an NICR, the tumor necrosis factor-alpha (TNF-alpha) response was similar and occurred on day 1, also pointing to normally functioning macrophages. However, the maximal appearance rates of interleukin-6 (IL-6) and IL-8 occurred later in the episodes with IICR compared to NICR (day 2 vs day 1, p < 0.05). No differences were found in vasodilating prostaglandins, mesothelial cell markers (cancer antigen 125, phospholipids, hyaluronan), and mesothelial cell numbers in the stable situation nor during peritonitis. Peritonitis can present as abdominal pain in the absence of a cloudy dialysate. In some of the patients this presentation occurred more than once. This impaired, most often delayed, cell reaction was associated with a delayed secondary cytokine response. As IL-6 and IL-8 can be synthesized by mesothelial cells, this suggests an impaired functioning mesothelium. This could not be confirmed, however, by a lower number of mesothelial cells in effluent or lower dialysate levels of mesothelial cell markers. PMID- 8728225 TI - Comparisons of Y-set disconnect system (Ultraset) versus conventional spike system in uremic patients on CAPD: outcome and cost analysis. AB - We conducted a single-blind, prospective randomized study on the use of the Y-set disconnect system (Ultraset) (U) versus the conventional (C) spike system to assess the peritonitis rate, exit-site infection (ESI), clinical outcome, the resulting hospitalization rate, and recurrent costs. Forty new end-stage renal failure patients admitted to the dialysis program were recruited into the study and 20 each were randomly allocated to the U and C systems. They were studied for a period of 12 months. The mean number of days required to train patients for the U and C systems were 8.6 and 9.8 days, respectively. The peritonitis rates for the U and C systems were one episode every 17 and 11.4 patient-months, respectively. The ESI rates for the U and C systems were one episode every 26.4 and 21.6 patient-months, respectively. Four catheters were removed due to fungal peritonitis (three with the C system and one with the U system). As related to peritonitis, patients on the C system required 57 hospital-days while those on the U system required 28 days per year. On cost analysis, the extra cost required for the U system can be offset by the other expenses incurred for events related to more infections on the C system. It is concluded that for the similar cumulative costs required for the patients on the two systems, the Y-set disconnect has a better morbidity profile than the conventional spike system. PMID- 8728226 TI - Health-care economics and peritoneal dialysis. AB - There is no question that the next decade will see an enormous growth in the worldwide dialysis population. Increasingly, old and sick patients will be accepted on dialysis, and the worldwide dialysis population characteristics will approach those now seen in the U.S. In addition, recent attention in the U.S. and elsewhere on the mortality of dialysis patients is already resulting in an improvement in this regard, further expanding the future dialysis population. PD will continue to increase in use, particularly with continued worldwide economic pressures on dialysis providers. To the extent that the efficacy of this treatment can be improved, without eroding its economic benefits over HD, it should fare well in the future. If, on the other hand, expensive "adjustments" in the practice of PD are needed to assure reasonable medical outcomes, PD will likely fade in popularity. The challenge to those involved in PD--clinicians, scientists, companies--is to develop a PD system that optimizes patient medical and psychosocial outcomes, and minimizes costs. If this cannot be accomplished, PD is likely to be replaced by new dialytic systems in the future (e.g., daily, slow, home hemodialysis) that can succeed in these areas. PMID- 8728227 TI - Socioeconomic and health-care policies and peritoneal dialysis. AB - 1. All ESRD treatments is expensive and a modality change adds considerably to that expense. 2. Ideally, the cost of ESRD therapy should be reported as the cost incurred for all aspects of the treatment to the health-care system. 3. Most studies show peritoneal dialysis to be less expensive than hemodialysis, but this can vary depending on the local costs of supplies and labor. 4. Strategic decisions with regard to new innovations should be made after a full analysis of all of the involved costs and savings that the innovation might introduce. 5. Economic factors and reimbursement rates appear to have an important impact on modality selection. PMID- 8728228 TI - Peritoneal dialysis in Asia. AB - The socioeconomic status of Asian countries is diverse, and government reimbursement policies for treatment of patients suffering from end-stage renal disease (ESRD) vary greatly from one country to another. Both of these factors have a major impact not only on the choice of treatment for ESRD but also on the utilization of peritoneal dialysis (PD) in this region. Based on the data collected from 11 representative Asian countries, several observations can be made. First, the treatment rates for ESRD in these countries correlated closely with their gross domestic product (GDP) per capita income. Second, the PD utilization rate appeared to have a biphasic relationship with the GDP per capita income and treatment rate, in that countries with the highest and the lowest treatment rates tended to have lower PD utilization rates, whereas countries with modest treatment rates tended to have higher PD utilization rates. The reason for low PD utilization in countries with the highest treatment rates differs from that in countries with low treatment rates. In the former, because of full government reimbursement, there is little physician incentive to introduce PD as an alternative form of ESRD treatment to in-center hemodialysis (HD), whereas in the latter, the complete lack of government reimbursement prevents the introduction of PD as a form of treatment. This pattern is likely to change in the future because, of the 11 countries surveyed, all except Thailand have recorded a growth rate which is higher for PD than HD over the last three years. The rate of utilization of different PD systems varies greatly among different Asian countries. Automated PD has yet to gain popularity in Asia. Conventional straight-line systems remain the dominant PD systems in use in Hong Kong, Korea, Thailand, and the Philippines, while in Malaysia and Singapore UV germicidal connection devices are most popular. However, in all these countries there has been a progressive shift over the last three years from the straight-line systems with or without germicidal connection devices to the disconnect systems. In China and India, where PD has been introduced only recently, the disconnect systems are used almost exclusively. The disconnect systems are also the most popular systems in use in Japan and Taiwan. As data concerning the cost-effectiveness of different PD systems becomes available, it is likely that trend towards a more liberal use of disconnect systems will continue in the future. The usage of low calcium peritoneal dialysate and the average number of daily CAPD exchanges also vary among the Asian countries. Low calcium peritoneal dialysate has been introduced only in Japan, Hong Kong, and Singapore, with the highest utilization rate (90%) recorded in Singapore. The Philippines had the lowest average number of daily peritoneal exchanges (6L) among the countries surveyed, followed by Hong Kong (6.4L), China and Indonesia (7L), and the rest (8L). The use of a lower number of exchanges was introduced in some countries, initially, mainly as a cost saving measure based on the assumption that Asians are of small body build. The justification for the continued use of a lower number of exchanges among Asian patients is debatable, but is supported by the acceptable, long-term clinical outcome of patients given this form of dialysis prescription. It is suggested that long-term prospective studies on dialysis adequacy and clinical outcome should be done in different ethnic groups in Asia to see if the similar guidelines with regard to dialysis adequacy can be applied uniformly to Orientals and Caucasians. PMID- 8728229 TI - Withholding or withdrawing dialysis: some ethical considerations. PMID- 8728230 TI - Integration of peritoneal dialysis in active uremia treatment. PMID- 8728231 TI - Quality-of-life assessment in patients on peritoneal dialysis: a review of the state of the art. PMID- 8728232 TI - Treatment of sexual dysfunction. PMID- 8728233 TI - Quality of life elderly and diabetic patients on peritoneal dialysis. PMID- 8728234 TI - Survival on chronic peritoneal dialysis: have results improved in the 1990s? AB - One hundred and seventy patients were treated with home peritoneal dialysis (PD) in our unit between 1986 and 1994. During this time lapse, several technical improvements were included in our practice. Among others there were: Swan neck permanent catheters, Y-systems, and automated home PD (APD). We reviewed our experience, to assess if these improvements had any impact on patient and technique survival, comparing patients who started PD between 1986 and 1989 (group A), with those who started PD between 1990 and 1994 (group B). Both groups had a comparable basal comorbidity, except for a higher proportion of elderly patients in group B (mean age 48 vs 58 years, p < 0.01). The incidence of peritonitis was lower in group B, while there were no differences in the rates of catheter-related infection or hospital admission. Also, there were no significant differences in patients or technique survival. The increasing presence of elderly patients in our PD unit was, apparently, determinant for the evolution of patient survival. On the other side, technical improvements had a marginal impact on technique survival. A good general PD survival in both groups, with few patients changing to hemodialysis (HD), may explain the lack of significant differences. In addition, peritonitis and inadequate PD/ultrafiltration (UF) were replaced by abdominal surgical events and social reasons as the main causes for PD failure in the second phase of the study. PMID- 8728235 TI - 213 elderly uremic patients over 75 years of age treated with long-term peritoneal dialysis: a French multicenter study. AB - We report our experience in 213 elderly patients over 75 years treated by peritoneal dialysis (PD) as first and exclusive dialysis therapy. The mean age at start of PD was 79.4 +/- 3.6 years, and the cumulative time on PD was 4551 months (mean time: 21.4 +/- 19.8 months). Twenty-six patients lived in institutions and 187 lived at home. Thirty patients had an effective autonomy with the ability to carry on normal activities. One hundred and two patients were cared for by a private nurse at home, and 46 patients were cared for in a family environment. Most cases were treated by three exchanges per day (152 cases) and used a nondisconnect system (175 cases) on account of absence of autonomy. The rate of peritonitis per patient-month was one episode per 16.8 patient-months. Patient survival (Kaplan-Meier curves) was 74%, 59%, 45%, and 19% at one, two, three, and five years, respectively. The causes of death were various with a higher frequency of cardiovascular causes (48.3% of the 116 deaths). Thirty-three patients died in less than six months including 18 patients in less than three months. IN CONCLUSION: elderly uremic patients can be treated with long-term PD with relatively good results. Mortality is high but essentially due to age and poor general status-the dedication of private home nursing is very important in treating elderly PD patients. This fact often is a necessary condition in maintaining these elderly patients at home. PMID- 8728236 TI - Muscle weakness in continuous ambulatory peritoneal dialysis patients. PMID- 8728237 TI - Hepatitis C antibodies in patients on peritoneal dialysis: prevalence and risk factors. AB - Our objective was to determine the prevalence of the antibody to hepatitis C (anti-HCV) in a population of end-stage renal failure patients on continuous peritoneal dialysis (CPD) and study the possible risk factors associated with anti-HCV seropositivity and seroconversion. A cross-sectional study included 155 adult patients enrolled in the CPD program in a single renal unit of a teaching hospital who were screened for anti-HCV by second-generation enzyme immunoassay, which was confirmed by recombinant immunoblot assay. Serum was also assayed for hepatitis B surface antigen (HBsAg). History of renal transplantation, blood transfusions, and exposure to hemodialysis was obtained from medical records. Ten of 155 patients (6.5%) in this study population were anti-HCV positive [anti HCV(+)] and 11/155 (7.1%) were HBsAg positive; no patient was positive for both. All the anti-HCV(+) patients were on continuous ambulatory peritoneal dialysis (CAPD); no continuous cycling peritoneal dialysis (CCPD) patient was anti-HCV(+). Exposure to hemodialysis was a risk factor for anti-HCV seropositivity, with 7 out of 10 (70%) anti-HCV(+) patients having been on hemodialysis compared to 55/134 (41%) anti-HCV(-) (p < 0.05, Fisher's exact test). No difference was noted between anti-HCV(+) and anti-HCV(-) groups in relation to age, gender, duration on CPD, renal transplantation, or exposure to blood transfusions. Seroconversion occurred in only one patient after a mean observation period of 20 +/- 0.6 months. The prevalence of anti-HCV seropositivity in this population of CPD patients is 6.5%, and HBsAG 7.1%. Exposure to hemodialysis is a significant risk factor for development of anti-HCV seropositivity. Seroconversion rate appears to be low. PMID- 8728238 TI - Sleep apnea in ESRD patients on HD and CAPD. PMID- 8728239 TI - Imaging of the peritoneal cavity in CAPD. AB - The aim of our study was the use of the optimal imaging of the peritoneal cavity (PC) for continuous ambulatory peritoneal dialysis (CAPD)-related problems. Ultrasonography (USG), direct abdominal radiography (DAR), peritoneoscintigraphy (PSG), and standard peritoneo-computed tomography (PCT) with reconstruction (R PCT) were performed in 25 patients on CAPD from three to 44 months. Studies were done at the beginning of CAPD (1-3 months) as well as in the noncomplicated and complicated course of CAPD. Group 1 comprised 17 patients in whom 77 PC images were taken in the non-complicated course of CAPD. Group II comprised 15 patients in whom 65 images were taken during or after complications. For USG and DAR we used standard equipment, PSG was done with sulfur colloid labeled with technetium 99m (Tc 99m), PCT, and R-PCT were done with Omnipaque and Somatom HiQ Siemens unit. In PCT, two- and three dimensional reconstruction were done by our own computer program. USG was recommended for imaging of tunnel infections, exit-site infection (ESI), and adhesion. PSG was useful in almost all observed complications of CAPD except thickening of the peritoneal membrane (PM). Standard PCT with R-PCT was more useful than PCT because of a more legible image of the peritoneal cavity, which gives the possibility of monitoring fluid distribution and measuring of intraperitoneal fluid volume. PMID- 8728240 TI - PD in Europe: nursing issues. PMID- 8728241 TI - Peritoneal dialysis in the Pacific. AB - Peritoneal dialysis (PD) in the Pacific is the predominant dialysis modality for home dialysis patients. Patient location, age, personal preference, and medical condition are all taken into consideration. However, with the steadily increasing number of patients on PD, financial constraints are now affecting the nurse patient ratios. As a consequence, PD nurses are constantly seeking new and improved clinical practice regimes with which they can provide and maintain quality, cost-effective nursing care. To enable PD nurses to provide such care, they are now, more than ever, involved in patient, professional, and management issues. These patient issues include, but are not limited to: the elderly, the diabetic, the indigenous patient, and their carers, peritonitis, exit-site infection, and adequate dialysis. Professional issues include the development of standards of clinical practice, nursing research, quality improvement, and tertiary education. Management issues are constantly encroaching into the clinical practice setting, therefore the PD nurse must now justify spending, develop strategic plans, and meet annual budgets. In conclusion, it can be seen that PD nurses in the Pacific are faced with similar patient-care, professional, and management issues as their counterparts throughout the world. PMID- 8728242 TI - Planes, kangaroos, and the CAPD manual. AB - The Western Australian (WA) Remote Area Dialysis Programme was developed in 1988 due to the cultural need to dialyze an increasing number of aboriginal patients in their own communities, rather than relocating them up to 3000 km away in Perth. The success of the program relies on remote area health services (RAHS), which have no prior experience in continuous ambulatory peritoneal dialysis (CAPD), providing consistent routine and emergency medical care to the patients. Our aim was to standardize the care of all CAPD patients in remote WA by providing the RAHS with an easy-to-follow manual. Although the RAHS received treatment protocols, and in-service education, consistent care was not always provided. We confirmed this by: (1) examining the existing quality assurance tools, peritonitis and hospital admission rates, (2) discussion with remote area staff regarding patients, and (3) informal assessment of remote area staff receptiveness to in-service education by a CAPD nurse. We identified the causes of the inconsistent care to be: (1) high remote area staff turnover (six months average for a registered nurse), (2) the protocols were difficult to follow, and (3) confusion for the RAHS as to the appropriate contact person at our hospital. In 1994, the situation was exacerbated by the dramatic increase in the number of patients and RAHS involved (14 new patients, bringing the total to 20 patients in 12 centers) plus the introduction of a second treating hospital (with differing protocols). A team of two CAPD nurses and two nephrologists was established, to collaborate with two remote area hospitals and the second treating hospital to produce the "Remote Area CAPD Manual." The manual is an easy-to-follow, step-by step guide for the management of CAPD by non-dialysis personnel. It has led to improved management of CAPD, improvement in communication with RAHS, and the increased confidence of remote area staff in the management of CAPD patients. In conclusion, RAHS can give consistent care if provided with clear, concise guidelines. PMID- 8728243 TI - A survey of the status of peritoneal dialysis nursing in Japan. PMID- 8728244 TI - Current nutritional recommendations in peritoneal dialysis. PMID- 8728245 TI - Nutritional status in continuous ambulatory peritoneal dialysis patients. PMID- 8728246 TI - Complications in peritoneal dialysis: peritonitis and exit-site infections. PMID- 8728247 TI - Peritoneal dialysis catheter complications. AB - A functional peritoneal dialysis catheter is essential for continuous ambulatory peritoneal dialysis to be an effective form of dialysis. The most common mechanical problems that hinder adequate function include obstruction, leaks, hernias, hemoperitoneum, and pain. The reported incidence of mechanical problems varies from 12% to 73%. Symptoms, diagnostic tests, risk factors, and treatment strategies are reviewed. Surgical intervention and catheter replacement are often required to resolve mechanical problems. Avoidance of mechanical problems reduces the risk of interruptions in peritoneal dialysis therapy. PMID- 8728248 TI - Psychosocial complications in PD. PMID- 8728249 TI - A home visit program for CAPD. AB - In the U.K. in the last five years a reassessment of the central role of the hospital has taken place, with a fundamental shift in care for people with increased dependencies to community settings. For patients using peritoneal dialysis a need has been realized for a home-visiting service by specialist renal nurses who can manage and coordinate the program of care, thus reducing the strain on hospital resources. In addition, the provision of a community nurse to visit the patient at home means that ongoing education and training can be undertaken in a suitable learning environment. The home-visiting program creates an opportunity to develop an holistic care plan using strategies of general health promotion as well as practical nursing care. This paper highlights the experiences of a pioneering model of community continuous ambulatory peritoneal dialysis care in the South London. Auditing of this unusually funded post has shown significant reductions in peritonitis, exit-site infections, and hospital admissions in this growing patient population. PMID- 8728250 TI - The use of community nurses for home peritoneal dialysis: is it cost-effective? AB - In the late 1970s, continuous ambulatory peritoneal dialysis was first introduced as a mode of treatment for patients with end-stage renal disease. Since that time many patients, themselves or with the help of family members, have routinely performed the therapy at home. There are now 2935 home peritoneal dialysis patients in Canada (36% of the total dialysis population). Today, however, the average patient on dialysis is likely to be older and have other complicating illnesses; moreover, patients may live alone, or have family members who are working. Over the past three years, through the use of innovative assistive devices and strong educational links with community nursing agencies, we have been able to manage peritoneal dialysis patients with complex needs in the home. We performed a retrospective analysis of 18 patients, with severe comorbid conditions, who were managed in the home with the help of community nurses. We will show that this is an economic, efficient, and effective method of caring for home dialysis patients with severe disabilities. Home care agencies need our support so that they can continue to help us manage the complex peritoneal dialysis patient in the 1990s. PMID- 8728251 TI - The CAPData computerized data system: a valuable communication and nursing tool. PMID- 8728252 TI - Nursing involvement in research: an academic pursuit or relevant to nursing practice? AB - It is said that nursing is 'informed by research'. We evaluated the effect a clinical research project had on the nursing procedure of a nonrotating, permanent continuous ambulatory peritoneal dialysis (CAPD) team involved primarily in patient care. A trial of the overnight solute clearance as a simple measure of the adequacy of CAPD was carried out and its effect on the following aspects of nursing practice was assessed: 1. Teamwork-Can research be successfully incorporated within the team's work structure in a clinical environment? 2. Nurse's role-Duties of the nurse involved in the research (selection of patients, organization of appointments, collection of samples, and calculation of results). 3. Nurse/patient relationship-Individual knowledge of patients' needs and communication skills employed (necessity for true informed consent). The experience showed us that the participation in research is not only possible, but also interesting from the nurses' viewpoint and of direct benefit to the patients. PMID- 8728253 TI - Successful pregnancy complicated by peritonitis in a 35- year old CAPD patient. AB - A 35-year old woman conceived six months after initiating continuous ambulatory peritoneal dialysis (CAPD). A medical plan was developed to give the patient adequate dialysis for a 1.5 g/kg/day protein intake. In addition, alterations in calcium, magnesium, and erythropoietin administration were required to reach the objectives set by the obstetrical/renal team. Three weeks prior to delivery, an amniotic leak developed, and vaginal cultures were positive for Escherichia coli. Oral amoxicillin was administered (500 mg per os q.i.d.) until the day of delivery. A 1545-g baby girl was delivered by cesarean section at 32 weeks. Five days postpartum the patient developed severe peritonitis, which subsequently grew E. coli. The patient fully recovered from the peritonitis, but catheter removal was required. Successful pregnancy can be expected on CAPD, and adequacy can be achieved with aggressive dialysis. Cesarean section delivery should probably be accompanied by full peritonitis therapy. PMID- 8728254 TI - Cardiovascular risk factors and their management in patients on continuous ambulatory peritoneal dialysis. PMID- 8728255 TI - CAPD--a risk factor in renal transplantation? ARF after transplantation. PMID- 8728256 TI - Low calcium dialysate and hyperparathyroidism. AB - A low calcium dialysate reduces hypercalcemia from calcium-containing phosphate binders and makes phosphate control possible without the use of aluminum salts. We asked whether this might, however, lead to hyperparathyroidism. We prospectively studied serum concentrations of parathyroid hormone levels (by an immunoreactive intact molecule assay) in 173 patients on continuous ambulatory peritoneal dialysis (CAPD) who were started on a low calcium dialysate (Ca2+ 1.25 or 1.00 mmol/L) because of hypercalcemia. Median follow-up was 13.2 months (range 1-28). Initial serum parathyroid hormone was [median(range)]: 70(5-1043) ng/L pre low calcium dialysate, and this rose to 130(5-914) ng/L at 0-6 months; 130(5 1030) ng/L at 6-12 months; 170(170-1400) ng/L at 12-18 months; and 130(5-1200) ng/L at 18-24 months (p = 0.0006). Twenty-two patients required a parathyroidectomy because of a sustained rise in parathyroid hormone that was not responsive to alfacalcidol and hypercalcemia. Initial serum parathyroid hormone was significantly higher in these patients at 359 (5-1073) ng/L as compared to a level of 69.5 (6-1147) ng/L in patients who did not have a parathyroidectomy (p = 0.0009). There was a significant sustained fall in mean serum corrected calcium from 2.77 (2.37-3.51) mmol/L to 2.53 (1.39-3.20) mmol/L at three months (p = 0.0006), a nonsignificant rise in mean serum alkaline phosphate from 179 (47 1858) mmol/L to 191 (55-1821) mmol/L (p = 0.15), and a fall in mean serum phosphate levels from 1.87 (0.59-3.18) mmol/L to 1.68 (0.45-3.6) mmol/L (p = 0.76). Our data suggest that the benefits of a low calcium dialysate in CAPD patients are balanced by an increased risk of hyperparathyroidism, and that this risk is higher in patients with an initially high serum parathyroid hormone level. PMID- 8728257 TI - The ethics of infant dialysis. AB - The proper treatment of an infant with end-stage renal disease depends upon a number of factors including parental willingness to take on the task, experience of the health-care team, local and regional resources, and society's willingness to accept this support as a standard of care. Whereas the ability to keep infants alive on peritoneal dialysis (PD) is obtainable, it is not without physical, financial, as well as emotional cost. In order for a family to agree to take on such a task, an understanding of the risks and long-term prognosis should be offered. This "informed consent" is difficult to obtain in such a highly charged situation when emotions often dictate choice independently of logic. Long-term outcome of infants on PD has improved over time, yet is still fraught with complications. Options of treatment or nontreatment are explored. PMID- 8728258 TI - Acute peritoneal dialysis access in infant renal failure. AB - Rapid onset of acute peritoneal dialysis (PD) in the infant population is, in part, dependent upon equipment availability. From March 1991 until June 1995, over 200 children at the University of Michigan have undergone dialysis for acute renal failure. Of these children, 29 infants (mean +/- SEM)(age 4.5 +/- 1.3 months; weight 4.8 +/- 0.5 kg) have had placement of an acute 5 French Cook PD catheter for dialysis. Complications including inadequate inflow in one case, bleeding in one case, and accidental removal in one case were infrequent. Duration of the placed catheters was 9.9 +/- 2.7 days, without the problems associated with chronic placement of a stiff catheter. Onset of dialysis occurred within minutes due to rapid access. We conclude that the placement of a 5 French Cook acute PD catheter for acute PD in the infant population is easily performed with minimal risk. Moreover, this allows for more rapid onset of PD when conditions mandate this need. PMID- 8728259 TI - Peritoneal dialysis prescription for neonates. AB - Despite the major advances in the management of renal failure, the mortality and morbidity rates remain higher in neonates than in older children. But, in most of the cases, effective peritoneal dialysis is possible, giving these neonates a chance of life. In infants and neonates, peritoneal permeability seems to be in a hyperpermeable state, perhaps only related to the volume of the instilled dialysate. Practical peritoneal prescriptions in neonates have to be balanced between either short dwell times optimizing ultrafiltration or long dwell times optimizing the diffusive process and blood purification. The recommendations for peritoneal dialysis in neonates usually admitted are short dwell times, one hour or less, adapted to ultrafiltration need, and small instilled dialysate volumes, 10-20 mL/kg initially, to avoid raised intra-abdominal pressure. Age-related particular risks are hyponatremia and hypophosphatemia. PMID- 8728260 TI - Intraperitoneal amino acids in children receiving chronic peritoneal dialysis. PMID- 8728261 TI - Tube feeding in infants on peritoneal dialysis. PMID- 8728262 TI - Nasogastric tube feeding in infants on peritoneal dialysis. AB - Enteral nutritional support is an important component of the care provided to infants receiving long-term peritoneal dialysis. In the majority of published experiences on this subject, the use of the nasogastric tube has facilitated the provision of required calorie and protein intake and resulted in an improved patient outcome. Advantages of the nasogastric route of nutritional support include the ease of administration, while recurrent emesis remains the most troublesome and frequent short-term complication associated with its use. Impaired oral-motor development may also result from nonoral feeding and should be addressed throughout the course of tube feeding. PMID- 8728263 TI - Nutritional status in children receiving chronic peritoneal dialysis. AB - Chronic peritoneal dialysis (CPD), widely used in uremic children, may have contrasting effects on the nutritional status of patients. Metabolic and nutritional abnormalities due to the combined effects of uremia per se, glucose absorption from the dialysate and catabolic factors, such as protein and amino acid losses into dialysate, poor appetite, and recurrent episodes of peritonitis are the most important. Although CPD allows for fewer dietary restrictions and supplies an extra amount of calories by glucose absorbed with the peritoneal fluid, when protein and energy intakes are assessed the protein intake was almost sufficient or more than that prescribed, whereas the energy intake was low. In CPD children the standard deviation score for weight, height, triceps skinfold thickness, and midarm circumference has been reported as more severely impaired in children less than ten years old. Anthropometric parameters did not worsen during CPD treatment. Plasma proteins and albumin are reported as being low in CPD children. The dietary intake and protein losses have been considered to be the most important determinants of the albumin level in CPD patients. The reported average dialysate losses of free amino acids (AA) during CPD in children vary from 0.02 to 0.03 g/kg/day in different studies. The patterns of plasma AA in CPD is represented by reduced levels of branched chain AA and of other essential amino acids and increased concentrations of some nonessential AA. Several factors may influence plasma AA profile: uremia per se, hormonal alterations, protein and AA losses, and dietary intake. A more specific uremic AA pattern is found in muscle, the largest pool of free AA in the body. Studies on muscle AA in adults on CPD are conflicting: some authors have reported several muscle AA alterations, but others have shown an almost normal pattern. Low valine and leucine muscle levels have been reported in children on CPD. PMID- 8728264 TI - Peritoneal equilibration in children. PMID- 8728265 TI - Performance and interpretation of the peritoneal equilibration test in children. PMID- 8728266 TI - Infectious complications in pediatric patients treated with chronic peritoneal dialysis (CPD). PMID- 8728267 TI - Psychosocial burdens of dialysis treatment modalities: do they differ and does it matter? PMID- 8728268 TI - Home health and respite care. AB - Families with a child on chronic peritoneal dialysis have to assume a significant burden of care, which can result in stress and potential complications. Adequate preparation of the child and family with appropriate information in many forms is essential. We describe a home-visiting strategy using the named nurse, dietitian, and social worker, which also includes liaison with nurseries, schools, community nurses, and primary care physicians. Respite care is provided by an occasional evening baby-sitting service and increasing use of daytime support from a home care nurse. The holiday support strategy includes a holiday caravan for families in addition to an organized holiday for children alone. A parents' support group has provided valuable feedback on quality-of-care issues. PMID- 8728269 TI - Intellectual output of children with chronic renal failure on continuous ambulatory peritoneal dialysis. AB - The aim of this study is to quantify the intelligence output in our pediatric population on continuous ambulatory peritoneal dialysis (CAPD) treatment. A total of 30 children were studied, with an age range of eight to 18 years. For evaluating a global intelligence quotient (IQ), the Wechsler test was applied according to their age. And, as a complement, a Bender test was also requested to deny or confirm brain damage. The Wechsler test showed an average intelligence quotient in most of the children (76.7%); a small group (16.7%) was classified as dull normal, 1 child had mild retardation, and 1 was borderline. All of them had a Bender test that did not correlate with brain damage. Most of them maintained a very high verbal IQ, but, when the performance IQ was qualified, 34% got a low score, and a certain difficulty in solving this part of the test was observed. Maybe this was influenced by chronicity of the sickness and/or the blood urea nitrogen level. In conclusion, the child's plasticity is his best quality to cope with uremia and the alterations caused by it. Therefore the idea is to consider the intelligence quotient as the capacity that the subjects have to use their own resources to cope with their environment. And, even though the neurological alterations exist, the child's plasticity helps him/her maintain a good global intelligence quotient, even though he/she is not having immediate transplantation. PMID- 8728270 TI - Determination of individual ultrafiltration time (APEX) and purification phosphate time by peritoneal equilibration test: application to individual peritoneal dialysis modality prescription in children. AB - Efficiency of peritoneal dialysis (PD) is dependent on adequate ultrafiltration (UF) and purification (solute clearance). These two goals apparently seem to conflict in terms of duration of dwells: short dwell time enhances UF capacity and, conversely, long dwell time enhances solute clearance. Peritoneal equilibration test (PET) allows an approach to the ultrafiltration time: the point at which the over time dialysate urea saturation and glucose desaturation curves cross, call APEX time. PET also allows an approach to the purification time: the point at which dialysate-to-plasma (D/P) concentration ratios over time are high. Because of the value of phosphate as a uremic factor of morbidity, we have chosen the time at which D/P phosphate is equal to 0.6 as a purification phosphate dwell time (PPT). A total of 17 patients were studied, over a five-year period, allowing 142 determinations. APEX times (range 18-71 min) and PPT (range 105-238 min) were spread over a wide distribution. PPT and APEX times were significantly shorter in children younger than three years of age than in children older than ten years of age. PPT were nearly four times longer than APEX times. Knowledge of these conflicting ultrafiltration and purification times should help, in our view, in the individual choice of the PD modality: if UF is the major goal, short dwell times should be used (automated PD); if purification is the major goal, long dwell times should be used, as in continuous ambulatory peritoneal dialysis; if both are the target goal, tidal PD should be discussed. PMID- 8728271 TI - Body composition of children on CAPD. AB - Our objectives were to estimate the body composition of children on continuous ambulatory peritoneal dialysis (CAPD), assess the fat-free mass (FFM) by bioelectrical impedance (BIA) and skinfold anthropometry (SF), and determine the effect of various parameters of the nutritional status and adequacy of dialysis on body composition. The design was a noninterventional retrospective clinical trial. The percent of fat ranged from 10% to 25% (15 +/- 2), when it was calculated from SF, and was from 9% to 32% (18 +/- 2) when it was calculated from BIA. There was a significant correlation between the percent of fat calculated by SF and by BIA (r = 0.94, p < 0.0001 with limits of agreement 4.16 to 1.37 and 0.40 to -3.19). The water content of FFM ranged from 70%-79%(74 +/- 1). There was a weak but significant correlation (r = 0.64, p = 0.016) between protein catabolic rate (PCR) and KT/V (V = 60% of weight). This correlation became more significant (r = 0.83, p = 0.0007) when the V of KT/V was calculated from BIA. The prediction of body composition with the use of bioelectrical impedance is a simple and reliable technique. Serial measurements of BIA might be an important tool for the assessment of the nutritional intervention. PMID- 8728272 TI - The influence of dialysate calcium concentration on the PTH level in children undergoing CAPD. AB - In 12 children aged four-and-a-half to 18 years (mean 11 +/- 4.2) undergoing continuous ambulatory peritoneal dialysis (CAPD), serum intact parathyroid hormone (iPTH), ionized calcium (iCa) levels, and calcium mass transfer (CaMT) were measured on three consecutive days: day 1, after a four-hour interval between dialyses; on day 2, after four hours dwell time with peritoneal dialysis (PD) Ca 3.5 mEq/L; and on day 3, after four hours dwell time with PD Ca 2.5 mEq/L. A significantly more negative CaMT was found when PD Ca 2.5 mEq/L was used, as compared with values obtained using PD Ca 3.5 mEq/L. Significantly lower parathyroid hormone (PTH) values were found after the interval between exchanges. We conclude that in order to properly evaluate parathyroid gland function and to decide whether or not to give vitamin D metabolites, a protocol for determining PTH should be standardized. PMID- 8728273 TI - Complications linked to chronic peritoneal dialysis in children after kidney transplantation: experience of the Italian Registry of Pediatric Chronic Peritoneal Dialysis. AB - Our objective was to evaluate the infectious complications of the post-transplant period attributable to the persistence of catheter and other complications when chronic peritoneal dialysis (CPD) was performed post-transplantation. The design was a retrospective study, and the setting was an Italian registry of pediatric chronic peritoneal dialysis. There were 86 pediatric renal transplants (9/86 from living related donors, 2/86 simultaneous liver and kidney transplantation for oxalosis). Six of 86 transplants were lost at follow-up. Mean age of the children (n = 80) at transplantation was 9.3 years (range: 1.7-21 years). They had been on CPD for a mean period of 1.7 years (range: 0.2-4.6 years). During CPD, 67 peritonitis episodes (80% related to exit-site and/or tunnel infections) were observed, with an incidence of peritonitis of one episode per 16 months CPD. The mean safe interval of peritonitis and/or exit-site or tunnel infection was 208 days (range: 36-1897 days). The mean time of catheter removal was 80.3 days (range: 0-216 days) post-transplantation. During the first month post transplantation, one episode of peritonitis secondary to a sepsis occurred in one child. No other episodes of peritonitis or exit-site and/or tunnel infections were observed. Two of 80 children returned to CPD (at four and at 12 months, respectively) because of persistent allograft failure. Furthermore, 12 patients were on CPD because of temporary graft failure. In all these patients the pretransplant peritoneal dialysis (PD) catheter was utilized, with no complications. These data show that the persistence of the PD catheter after kidney transplantation has produced no infections or other complications. What is more, the catheter was safely utilized during acute rejection or primary allograft nonfunction. PMID- 8728274 TI - Comparison of patient hospitalization in chronic peritoneal dialysis and hemodialysis: a pediatric multicenter study. AB - Patient hospitalization was compared in 207 pediatric patients (age < or = 15 years at the start of dialysis) on chronic peritoneal dialysis (CPD) (127 patients) or center hemodialysis (HD) (80 patients), treated in 17 dialysis centers during the period 1989 to 1994, and followed up for at least three months. The hospitalization rate was expressed as hospital days per patient month, and was calculated on the overall period of treatment and separately for the first and second year. Since the age at start of dialysis markedly differed between CPD (8.2 +/- 4.7 years) and HD (11.2 +/- 2.9 years) patients (with no HD patient younger than five years), results are separately presented in three patient groups: CPD patients aged < 5 years (A); CPD patients aged five to 15 years (B); HD patients (C). The duration of hospitalization was subdivided according to the following different causes: routine (monitoring of dialysis adequacy), complications of the modality, patient primary renal disease, and other causes. The results are presented in Table 1. A statistically significant difference in total days hospitalized was found between each of the two groups of CPD patients and the HD patients; the results for hospitalization for dialysis related complications were higher in the group of younger children on CPD, while the difference between the two age-matched groups of patients on CPD and HD was not significant. PMID- 8728275 TI - Environment: the white noise of health. PMID- 8728276 TI - Frankenstein in the age of Prozac. PMID- 8728277 TI - What is a natural disaster? PMID- 8728279 TI - The whole story: nature, healing, and narrative in the Native American wisdom tradition. PMID- 8728278 TI - Imagining cancer: Sara Coleridge and the environment of illness. PMID- 8728280 TI - "A slap in the face for old Mother Nature": Disease, debility, and decay in Huysmans's A Rebours. PMID- 8728282 TI - Environmental wellness. PMID- 8728281 TI - "Another name for health": Thoreau and modern medicine. PMID- 8728283 TI - [Endothelial cell function and angiogenesis]. AB - Angiogenesis, a process of new blood vessel formation, is an integral part of development, wound repair and tumor growth. The formation of capillary networks requires a complex series of cellular events, in which endothelial cells locally degrade their basement membrane, migrate into the connective tissue stroma, proliferate at the migrating tip, enlongate and organize into capillary loops. In response to angiogenic stimuli, endothelial cells in culture develop networks of capillary-like tubes. In this paper, we showed the relationship between angiogenesis and diseases, the assay systems of angiogenesis and the reports of angiogenesis published recently. PMID- 8728284 TI - [Regulators of angiogenesis]. AB - Endothelial cells lining the lumen of vessels are maintained in the quiescent state and play important physiological roles. Yet, they can be de-differentiated and become one of the most rapidly proliferating of all cell types when stimulated. Angiogenesis or neovascularization is defined as the formation of new capillary vessels from existent microvessels, which plays a major role in the evolvement of a vascular supply in tissue during development or remodeling and disease. Angiogenesis is believed to be regulated by the balance between inducers and inhibitors. In this review article, I will summarize the molecules that regulate the process of angiogenesis. PMID- 8728285 TI - [Involvement of the VEGF-Flt-receptor family in angiogenesis]. AB - Angiogenesis is important not only in normal embryogenesis, tissue organization and its maintenance but also in pathological processes such as ocular disease in diabetes mellitus and rapid growth of tumors in vivo. Recently, endothelial cell specific growth factor (VEGF) and its receptors (Flt family) has been characterized, and this ligand-tyrosine kinase receptor is considered to be one of the most important systems involved in angiogenesis. VEGF is induced by a variety of normal or tumor cells under conditions such as hypoxia and hypoglycemia and in the presence of substances such as hormones and growth factors. On the other hand, receptors of the Flt family (Flt-1, KDR/Flk-1, Flt-4) are basically strictly expressed only on vascular endothelial cells with a rare exception. Thus, the stimulation of VEGF-Flt towards angiogenesis is through a paracrine mechanism. A direct involvement of Flt-1 and KDR/Flk-1 in vasculogenesis/angiogenesis has recently been demonstrated by gene targetting studies. Blocking of this system might be a useful tool for suppression of solid tumors in vivo. PMID- 8728286 TI - [Transforming growth factor-beta and its receptors]. AB - Transforming growth factor-beta (TGF-beta) is a family of multifunctional proteins that inhibit the growth of most cell types, and these proteins induce the deposition of extracellular matrix. TGF-beta inhibits the growth and migration of endothelial cells in vitro, but induces angiogenesis in vivo. TGF beta belongs to a larger superfamily known as the TGF-beta superfamily, which includes activins and bone morphogenetic proteins. TGF-beta is produced as latent high molecular weight complexes from producer cells and is then activated by plasmin or thrombospondin. Latent TGF-beta binding protein (LTBP) is a component of the latent TGF-beta complex produced from platelets and many other cell types; LTBP plays an important role for the interaction of the latent TGF-beta complex with extracellular matrix components. TGF-beta binds several cell surface receptors, including type III receptor (betaglycan), endoglin, type II receptor and type I receptor. The type III receptor and endoglin are indirectly involved in the signal transduction. The Type II and type I receptors have intracellular serine/threonine kinase domains. They form a heteromeric complex after ligand binding and are most important for signal transduction; the type II receptor transactivates the type I receptor, which transduces various signals. PMID- 8728287 TI - [Shear stress and vascular formation]. AB - Blood flow plays important roles in the morphogenesis of blood vessels. For instance, increases in blood flow induce dilatation of the blood vessels, while decreases in blood flow cause reduction of vessel diameter. Blood flow also stimulates angiogenesis. In these blood flow-dependent phenomena, wall shear stress generated by flowing blood that acts on vascular endothelial cells works as a key factor. Numerous in vivo and in vitro studies have demonstrated that mechanical forces, shear stress, actually modulate the morphology and many functions of endothelial cells, and these forces also alter their gene expression. More recently, a cis-acting shear stress responsive element was identified in the promoters of endothelial genes that respond to shear stress, suggesting a common mechanism linking biomechanical forces to gene expression. Details of the process in which shear stress-mediated changes in endothelial cell functions lead to vascular remodeling and angiogenesis, however, are not entirely clear. Elucidation of this problem will give us not only a better understanding of the morphogenesis of blood vessels but also new therapies that can help manage or prevent cardiovascular diseases including atherosclerosis. PMID- 8728288 TI - [Extracellular matrix components and angiogenesis]. AB - Angiogenesis, the formation of new capillaries, is critical for normal physiological processes such as embryonic development and wound repair. However, it also facilitates pathological processes including tumor growth, metastases, proliferative diabetic retinopathy and pannus formation in rheumatoid arthritis. It has been described that the angiogenesis occurs through a series of events that include endothelial cell protease production, migration and proliferation, tubule formation, and basement membrane incorporation. Within the last two decades, with in vivo assay systems, various kinds of growth factors were identified as angiogenic factors that promote endothelial cell proliferation and migration, while an in vitro model for angiogenesis indicated that extracellular matrix (ECM) proteins stimulated endothelial cells to organize into a capillary like tubular network and suggested that the ECM proteins are involved in the tubule formation process of antiogenesis. Recent papers reported the identification of the specific receptors on endothelial cells involved in the ECM induced capillary tube formation. This article will focus on papers describing the in vitro analyses of tube formation of endothelial cells. PMID- 8728289 TI - [Phenotypic change of the smooth muscle cell and atherosclerosis]. AB - Phenotypic change of the smooth muscle cell (SMC) is implicated in normal development as well as several pathological processes including atherosclerosis. In general, differentiation SMCs show contractile responses to different exogenous stimuli and are inactive in mitosis, while undifferentiated or dedifferentiated SMCs show a mitogenic response and are not contractible. In the present review, we describe structural and functional aspects of the phenotypic change of SMCs with special reference to their role in atherogenesis. SMCs derived from atherosclerotic intimal lesions (intimal SMCs) show more amplified growth potential and chemotactic activity than medial SMCs; and furthermore, they require a macrophage-like phenotype: uptake of modified low density lipoproteins through the scavenger receptor, which leads to high tendency toward foam cell formation. Platelet-derived growth factor, secreted from most of the cells existing in atherosclerotic plaques, is one of candidates that promote the formation of such a highly dedifferentiated intimal SMC. Clinical and experimental evidence supports the concept that an appearance of the pathological intimal SMCs is a key step for their abnormal proliferation in atheromatous lesions. Recent advances in characterization of the phenotype-specific molecular markers for SMC, such as myosin heavy chain, caldesmon, and calponin, are also described. PMID- 8728290 TI - Biochemical assessment of sympathetic activity and prejunctional modulation of noradrenaline release in humans. PMID- 8728291 TI - Endogenous ouabain, sodium balance and blood pressure: a review and a hypothesis. AB - OBJECTIVE: To assess possible relationships between endogenous ouabain, sodium balance and blood pressure. CONTENT: This review concerns the structure of endogenous ouabain, circulating levels of this steroid in various disorders of fluid and electrolyte balance, recent evidence for the association of endogenous ouabain with human hypertension, the influence of sodium and volume factors on ouabain-induced hypertension, and possible mechanisms for the hypertensinogenic activity of ouabain. CONCLUSIONS: The human circulation contains a closely related isomer of ouabain of putative adrenocortical origin. Elevated circulating levels of this 'endogenous ouabain' are common but not universal in physiologic and pathologic states associated with positive sodium balance or high blood pressure, or both. In the absence of adrenal hyperfunction, elevating circulating levels of endogenous ouabain appear to be secondary to impaired renal clearance. Prolonged elevation of circulating ouabain in the rat induces sustained hypertension. This model exhibits normal plasma renin activity, increased levels of ouabain in the hypothalamus, pituitary, and kidney, and responds to angiotensin converting enzyme inhibitor. In rats with normal kidney function, ouabain-induced hypertension is primarily sodium-insensitive although maneuvers that hinder renal sodium excretion augment the pressor effect of this steroid. Prolonged administration of ouabain into the brain ventricles augments sympathetic nervous system activity and induces sustained hypertension. These observations lead us to propose the following hypothesis. Among Caucasian patients with essential hypertension, a large fraction have elevated circulating levels of endogenous ouabain, possibly caused by an inherited or acquired renal defect in clearance of this steroid. In these patients, and in rats with ouabain induced hypertension, increased local generation of, or increased target organ sensitivity to, angiotensin II, or both, may contribute critically to heightened vasoconstriction and a sustained increase in blood pressure. Investigations of the efferent pressor mechanisms and the renal handling of endogenous ouabain are novel approaches to the etiology and therapy of several common cardiovascular disorders. PMID- 8728292 TI - Endogenous ouabain, sodium balance and blood pressure. PMID- 8728293 TI - The adrenal renin-angiotensin system: a local hormonal regulator of aldosterone production. PMID- 8728294 TI - Chronic infusion of low-dose angiotensin II potentiates the adrenergic response in vivo. AB - OBJECTIVE: The aim of this study was to investigate the role of angiotensin II induced potentiation of the alpha 1-adrenergic contractile response in the aetiology of low-dose angiotensin II-induced hypertension. METHODS: Wistar rats (250g) received angiotensin II (120 ng/kg per min) from subcutaneous minipumps for 21 days. The responses of vaso-active properties of second-order mesenteric arteries (200 micron) to potassium, phenylephrine, angiotensin II and acetylcholine were assessed. The acute amplification effects of angiotensin II on the response to phenylephrine were examined. RESULTS: Angiotensin II induced a progressive hypertension, which reached a plateau after approximately 5 days. The responses to potassium, angiotensin II and acetylcholine were not significantly modified in rats treated chronically with angiotensin II. The major finding of this study is that the response to phenylephrine (1-3 mumol/l) was potentiated (sevenfold at 1.75 mumol/l) after chronic treatment with angiotensin II. In control vessels acute addition of angiotensin II (10(-10) mol/l) produced no contraction but induced potentiation of the phenylephrine response (1-3 mumol/l). No further potentiation of the phenylephrine response was observed in the rats treated chronically with angiotensin II. CONCLUSIONS: Thus, although the direct contractile responses to potassium and angiotensin II remain unaffected following chronic angiotensin II treatment, the alpha 1-adrenergic contractile response to phenylephrine is significantly potentiated by angiotensin II in this model of hypertension. We suggest that this potentiation contributes to the hypertension observed in response to infusion of low-dose angiotensin II. PMID- 8728295 TI - Coronary vascular ATP-sensitive potassium channels are activated to a greater extent in spontaneously hypertensive rats than in Wistar-Kyoto rats. AB - OBJECTIVE: To determine whether opening of coronary vascular adenosine 5' triphosphate (ATP)-sensitive potassium (KATP) channels is involved in the maintenance of resting coronary flow in hypertrophied hearts. METHODS: We examined the effects of glibenclamide, a selective inhibitor of KATP channels, on basal coronary vascular tone in Wistar-Kyoto (WKY) rats and spontaneously hypertensive rats (SHRs). Using a Langendorff system, the hearts from WKY rats and SHRs were isolated and perfused with oxygenated Krebs-Henseleit solution at a constant perfusion pressure of 60 and 100 mmHg respectively. RESULTS: Basal coronary flow and myocardial oxygen consumption (MV02) were similar in SHRs and WKY rats. The percentage decreases in coronary flow with glibenclamide at graded doses were greater (P < 0.01) in SHRs than in WKY rats (n = 8), whereas the percentage decreases in MV02 with glibenclamide were similar in the two groups. The decreases in coronary flow caused by U46619 (a thromboxane A2-mimetic agent) were similar in SHRs and WKY rats (n = 4). The increase in coronary flow caused by pinacidil (a KATP opener) was greater in SHRs than in WKY rats; glibenclamide prevented the pinacidil-induced increase in coronary flow in both SHRs and WKY rats. There was a significant positive correlation between the glibenclamide induced decrease in coronary flow and the degree of left ventricular hypertrophy (r = 0.54, P < 0.05). CONCLUSION: Our results suggest that the basal opening state of coronary vascular KATP channels is activated to a greater extent in SHRs than WKY rats, which may contribute to the maintenance of basal myocardial perfusion in hypertrophied hearts. PMID- 8728296 TI - Nerve growth factor gene and hypertension in spontaneously hypertensive rats. AB - OBJECTIVE: High blood pressure in spontaneously hypertensive rat (SHR) is associated with increased sympathetic innervation of key tissues, possibly as the result of increased nerve growth factor (NGF). The aim of this study was to test for genetic linkage of the NGF gene to high blood pressure. DESIGN: We studied NGF gene expression in young SHR and examined linkage of the NGF locus to mean arterial pressure in genetically segregating crosses of SHR and normotensive Donryu (DRY) rats. METHODS: NGF mRNA was measured by Northern blot, and a restriction fragment length polymorphism of the NGF gene revealed after digestion with the NsiI restriction enzyme was used to study inheritance. RESULTS: Levels of NGF mRNA were detected easily in the kidneys of 2-, 4- and 10-week-old SHR but not in age-matched DRY rats. In an F2 population, the blood pressure of rats homozygous for the DRY NGF allele was 6 mmHg less than in heterozygotes and 8 mmHg less than in rats homozygous for the SHR NGF allele (analysis of variance, P < 0.004). In backcross rats the blood pressure of NGF heterozygotes was not significantly different from that of SHR homozygotes. CONCLUSION: These results indicate differences in renal NGF mRNA in SHR during the development of hypertension and suggest that a genetic locus in or near the NGF gene contributes in a Mendelian dominant pattern to a significant increment in blood pressure in SHR. PMID- 8728297 TI - Blunted renal vascular response to angiotensin II is associated with a common variant of the angiotensinogen gene and obesity. AB - OBJECTIVE: Recently, we reported evidence for genetic linkage between human essential hypertension and the angiotensinogen gene (AGT) and an association with a common molecular variant of this gene (methionine 235 --> threonine or T235). Other studies had led us to hypothesize that blunted renal plasma flow responses to infused angiotensin II (Ang II) when in high salt balance may reflect increased intrarenal formation of Ang II, a condition that might promote hypertension. Here we examine the relationship between AGT genotype and renal vascular response to infused Ang II. METHODS: Hypertensive (n = 34, all off medication) and normotensive (n = 57) members of families with a history of hypertension (age 18-60 years) as well as 29 normotensive volunteers without a family history of hypertension were studied after controlled diets with 200 mequiv./day sodium. Ang II was infused at a mildly pressor dose (3 ng/kg/min) and renal plasma flow was determined by steady-state plasma para-aminohippurate concentration. RESULTS: After correction for covariates in multivariate analyses, participants homozygous for the T235 variant had significantly diminished renal plasma flow responses to the Ang II infusion (P = 0.005). Changes in renal arterial resistance were also blunted in the T235 homozygotes. Similar results were found when analysis was restricted to normotensive participants or subdivided based on family history of hypertension. No confounding factors associated with AGT genotype that could explain these differences were found. Furthermore, obesity, which also suppressed renovascular response to Ang II, was found to interact significantly (P = 0.017) with genotype such that, among T235 homozygotes, obesity had a greater blunting effect on renal vascular response. CONCLUSIONS: Expected renovascular response to infused Ang II was blunted in persons with the AGT TT genotype. This is the first report of an association between a specific gene variant and altered renal physiology in humans with particular relevance to essential hypertension. PMID- 8728298 TI - Adrenomedullin as a novel antiproliferative factor of vascular smooth muscle cells. AB - OBJECTIVE: The present study was designed to examine whether adrenomedullin affects fetal calf serum (FCS)-stimulated proliferation in cultured rat vascular smooth muscle cells (VSMCs). METHODS: Rat VSMCs were grown from explants of Sprague-Dawley rat aorta and were grown using the standard cell culture method. After incubation for 24 h with various concentrations of adrenomedullin in the presence of 5% FCS, trichloroacetic acid-insoluble tritiated thymidine was measured in a liquid scintillation counter. After incubation for 48 h, cell counts were performed. Cyclic adenosine 3',5'-monophosphate (AMP) levels were determined by radioimmunoassay. RESULTS: Rat adrenomedullin exhibited concentration-dependent inhibition of the FCS-stimulated increase in thymidine incorporation between 10(-7) and 10(-9) mol/l and of cell number at 10(-7) mol/l. However, the calcitonin generelated peptide (CGRP) receptor antagonist human CGRP(8-37) abolished these antiproliferative effects of rat adrenomedullin. Inhibition by adrenomedullin of FCS-stimulated cellular proliferation was paralleled by an increase in the cellular level of cyclic AMP. 8-Bromocyclic AMP, a cyclic AMP analogue, and forskolin, an activator of adenylate cyclase, inhibited the FCS-stimulated increase in thymidine incorporation and cell number. CONCLUSIONS: These results suggest that adrenomedullin inhibits FCS-stimulated proliferation in cultured rat VSMCs, probably through a cyclic AMP-dependent process. Taken together with the finding that adrenomedullin is synthesized in and secreted from vascular endothelial cells, adrenomedullin may play a role as an antiproliferative factor for VSMCs in a paracrine fashion. PMID- 8728299 TI - Effects of benidipine, a calcium antagonist, on urinary kallikrein excretion and renal impairment in experimental diabetes. AB - OBJECTIVE: To assess the long-term effects of different antihypertensive agents on urinary protein excretion and kallikrein excretion in diabetic rats with renal impairment. METHODS: Uninephrectomized streptozotocin diabetic Wistar-Kyoto rats were randomly assigned to receive vehicle, a calcium antagonist (benidipine) or an angiotensin converting enzyme inhibitor (captopril) for up to 12 weeks. Active kallikrein was determined by its kininogenase activity, and generated kinins were measured by radioimmunoassay. Total kallikrein was also determined by measuring kininogenase activity after inactive kallikrein had been activated with trypsin. RESULTS: Urinary protein excretion increased significantly in diabetic rats compared with non-diabetic rats. Urinary active kallikrein excretion was significantly reduced in diabetic rats, whereas urinary total kallikrein excretion was unchanged, resulting in a reduced percentage of active to total kallikrein compared with that in non-diabetic rats. Benidipine and captopril reduced blood pressure and attenuated the development of diabetic renal impairment in a similar manner. However, only benidipine attenuated the decreases in urinary active kallikrein excretion and the ratio of active to total kallikrein in diabetic rats. CONCLUSIONS: Although pathophysiological relevance of impaired urinary kallikrein activation to the development of diabetic renal impairment remains to be determined, our result might suggest a new mechanism by which calcium antagonists protect the kidney from diabetic renal impairment. PMID- 8728300 TI - Microalbuminuria screening by reagent strip predicts cardiovascular risk in hypertension. AB - OBJECTIVE: We tested the hypothesis that qualitative microalbuminuria (MAU) screening in a practice setting would identify non-diabetic hypertensive patients at high risk of developing cardiovascular disease. DESIGN: We enrolled general practitioners throughout Germany, who obtained histories, physical examinations, and routine laboratory values as clinically indicated on treated or non-treated hypertensive, non-diabetic patients. MAU was measured with a albumin-sensitive, immunoassay test strip. We studied 11 343 non-diabetic hypertensive patients. RESULTS: The patients' mean age was 57 years, 51% were men and mean hypertension duration was 69 months. Twenty-five per cent had coronary artery disease, 17% had left ventricular hypertrophy, 5% had had a stroke, and 6% had peripheral vascular disease. MAU was present in 32% of men and 28% of women (P < 0.05). In patients with MAU, 31% had coronary artery disease, 24% had left ventricular hypertrophy, 6% had had a stroke, and 7% had peripheral vascular disease. In patients without MAU, these rates were 22%, 14%, 4%, and 5% respectively: lower in every category (P < 0.001). Further, in patients with coronary artery disease, left ventricular hypertrophy, stroke, and peripheral vascular disease, MAU was significantly greater than in patients who did not have these complications (P < 0.001). MAU increased with age, severity of hypertension and duration of hypertension, was associated with higher plasma creatinine values, and was more common in patients with hyperlipidemia (P < 0.05). CONCLUSION: On the basis of our survey, we conclude that qualitative MAU determinations identify hypertensive patients with particular cardiovascular risk in a practice setting. PMID- 8728301 TI - The relationship of blood pressure with glucose, insulin, heart rate, free fatty acids and plasma cortisol levels according to degree of obesity in middle-aged men. AB - OBJECTIVE: To investigate the relationships of blood pressure with carbohydrate metabolism, sympathetic activity and cortisol at different levels of body mass index in middle-aged men. METHODS: Cross-sectional data concerning men studied in the Paris Prospective Study I were analysed. The cohort included 6424 subjects aged 40-53 years at entry, who were not being treated for hypertension or diabetes and had no overt heart disease. The parameters analysed were glucose, insulin and free fatty acids levels, all assessed during fasting by the subject and 2 h after a 75 g glucose load, resting heart rate and morning plasma cortisol levels. RESULTS: Subjects with systolic blood pressure > or = 160 mmHg had significantly higher glucose concentrations at any body mass index level whereas the difference in insulin levels between the subjects with and without high systolic blood pressure increased with body mass index. Heart rate, free fatty acids level and cortisol level were significantly higher in men with high systolic blood pressure. However, these parameters showed significant decreasing trends with body mass index. In normotensives, no such trends were observed. When analysing data according to diastolic blood pressure, the limit being 95 mmHg, similar results were obtained for glucose and insulin levels, but no trend in heart rate, free fatty acids level and cortisol level with the body mass index level was statistically significant. CONCLUSIONS: Mean glucose concentrations are higher in hypertensive men at all body mass index levels, whereas relative hyperinsulinaemia is present only in the more corpulent hypertensives. Heart rate, free fatty acids level and morning plasma cortisol level are elevated in hypertensive subjects at any body mass index level, but particularly in the lean ones with high systolic blood pressure. PMID- 8728303 TI - Finapres tracking of systolic pressure and baroreflex sensitivity improved by waveform filtering. AB - OBJECTIVE: Arterial pressure waveforms distort between brachial and finger arteries, causing differences mainly in systolic pressure. Distortion, reportedly, can be removed by applying a waveform filter to the finger pressure. DESIGN: We analysed the data from two studies that detected discrepancies in systolic tracking between Finapres and brachial pressures. The first set comprised waveforms of seven volunteers during incremental bicycle exercise to exhaustion and the second set comprised waveforms of eight volunteers during increasing phenylephrine infusion. METHODS: We applied the filter and compared 1 min averaged unfiltered and waveform-filtered finger and brachial pressures. RESULTS: During exercise, finger systolic pressure overestimated brachial increasingly, from 7(SD 10) mmHg at rest to 27(17) mmHg at maximal exertion. Differences were reduced by waveform filtering from 3 (SD 9)mmHg at rest to 1 (SD 15)mmHg at maximal exertion. During phenylephrine infusion finger systolic pressure overestimated brachial pressure, but the magnitude of the overestimate decreased from 14 (SD 15)mmHg at baseline to -1(SD 16)mmHg at maximal rate. After waveform filtering overestimation was an almost constant 6(SD 11)mmHg. Median baroreflex sensitivities from brachial, unfiltered and waveform-filtered finger pressure were 5.8, 7.5 and 5.3 ms/mmHg and correlation increased after filtering. The results indicate improved systolic pressure tracking after waveform filtering. CONCLUSIONS: Finger pressure distortion follows a general pattern correctable by waveform filtering. Waveform filtering allows a 'brachial' view to be obtained from Finapres data. PMID- 8728302 TI - Insulin, hypertension and antihypertensive drugs in elderly patients: the Rotterdam Study. AB - OBJECTIVE: To investigate the association between insulin, hypertension and antihypertensive drug use in elderly patients. DESIGN AND METHODS: Blood pressure, use of antihypertensive drugs and glucose metabolism were examined in 5453 men and women (mean age 68.8 years). This was part of the baseline examination of the Rotterdam Study, a population-based study of chronic diseases in elderly patients. Serum insulin was measured 2 h after an oral glucose load. RESULTS: Patients with hypertension had a significantly higher mean post-load insulin level than those without: 71.3 mU/l versus 59.3 mU/l (P < 0.001, adjusted for age). Systolic blood pressure increased by 0.25 mmHg per 10 mU/l insulin (95% confidence interval 0.15-0.35, adjusted for age, sex and antihypertensive drugs), whereas the increase in diastolic blood was 0.07 mmHg per 10 mU/l (0.01-0.13). Whereas insulin resistance was higher in patients with hypertension, the increase in insulin resistance with age was much more apparent in normotensive patients, resulting in similar insulin levels at high ages. Those using antihypertensive drugs, however, had higher insulin levels at all ages. CONCLUSION: The results of this study show that hyperinsulinemia is associated with raised blood pressure in elderly people, and suggest that the age-associated increase in insulin resistance is diminished in patients with raised blood pressure. The use of antihypertensive drugs, however, appears to be accompanied by an independent additional increase in insulin resistance at all ages. PMID- 8728304 TI - Association of genetic variation with interindividual variation in ambulatory blood pressure. AB - OBJECTIVE: To determine whether genetic and non-genetic components of interindividual variation in systolic and diastolic blood pressure are constant throughout the day or are time or activity dependent. METHODS: We obtained 24 h ambulatory blood pressure recordings in 263 members of 68 unrelated nuclear families (i.e. parents and their offspring) representative of the Caucasian population of Rochester, MN, USA. Using the time each patient got into bed as a reference point, we identified 198 records in which this reference point was preceded by eight consecutive active hours (out of bed) and followed by four consecutive inactive hours (in bed) in which four or more blood pressure readings taken each hour were judged to be technically satisfactory. For each hourly mean for systolic and diastolic blood pressure, we estimated total interindividual variance, variance associated with concomitant variables (generation; sex within generation strata; and age, height, weight, body mass index, and abdomen-to-hip ratio within generation and sex strata), and variance associated with additive genetic effects (i.e. the chief cause of resemblance between relatives). To assess trends in each component of interindividual blood pressure variance over the 12 h period, we estimated the slope of the linear regression line fit to the hourly estimates. RESULTS: For systolic blood pressure, total interindividual variance did not change significantly (slope of regression line = -0.23, P = 0.717). In contrast, total interindividual variance for diastolic blood pressure was greater during active hours than inactive hours (slope of regression line = 5.53, P < 0.001). For both systolic and diastolic blood pressure, variance associated with the concomitant variables was greater during active hours than during inactive hours (for systolic blood pressure slope of regression line = 2.98, P = 0.001; for diastolic blood pressure slope of regression line = -6.14, P < 0.001). Likewise, for both systolic and diastolic blood pressure, variance associated with additive genetic effects was also greater during active hours than during inactive hours (for systolic blood pressure slope of regression line = -1.65, P = 0.090; for diastolic blood pressure slope of regression line = 1.47, P = 0.018). CONCLUSIONS: This study demonstrates that components of interindividual variation in blood pressure are not constant, but are time or activity dependent. PMID- 8728305 TI - Prediction of patient responses to antihypertensive drugs using genetic polymorphisms: investigation of renin-angiotensin system genes. AB - OBJECTIVE: To investigate whether the M235T polymorphism of the angiotensinogen (AGT) gene and the insertion/deletion (I/D) polymorphism of the angiotensin-1 converting enzyme (ACE) gene predict blood pressure response to different antihypertensive agents. DESIGN: Sixty-three patients with untreated essential hypertension were randomly assigned in a placebo-controlled crossover comparison to atenolol 50 mg once daily, lisinopril 10 mg once daily and nifedipine SR 20 mg twice daily, and the effect on blood pressure was assessed by ambulatory blood pressure monitoring (ABPM). In a further 44 patients, placebo-controlled ABPM data were available after treatment with a single agent (atenolol 50 mg once daily in 16 cases and lisinopril 10mg once daily in 28 cases). The change in systolic and diastolic blood pressure achieved by each agent was analysed for association with genotypes at the AGT and ACE gene loci. METHODS: Polymerase chain reaction (PCR) amplification of genomic DNA from each individual was used to identify the I/D polymorphism of the ACE gene. The M235T polymorphism of the AGT gene was detected by Tth111I digestion of PCR product. RESULTS: There was no significant association between response to any drug and either the AGT M235T or ACE I/D polymorphisms. CONCLUSIONS: The large variability between individuals in the observed blood pressure response to these agents cannot be attributed to the polymorphisms analysed at the ACE and AGT loci. PMID- 8728306 TI - Comparative effects of combination drug therapy regimens commencing with either losartan potassium, an angiotensin II receptor antagonist, or enalapril maleate for the treatment of severe hypertension. AB - OBJECTIVE: To compare the efficacy and safety of a regimen of losartan potassium (losartan) and a regimen of enalapril maleate (enalapril) in a randomized trial of patients with severe hypertension in which the initial treatments were blinded. DESIGN AND METHODS: Seventy-five patients, 23-74 years of age, with sitting diastolic blood pressure of 115-130mmHg, were enrolled in a 12-site multicenter study. The primary efficacy parameters were the change in trough systolic and diastolic blood pressure, as well as response to treatment in terms of categories of hypertensive response. RESULTS: A gradual reduction in mean sitting diastolic blood pressure was observed in all patients treated from week 1 to 12 (10-29mmHg for the losartan regimen and 14-32 mmHg for the enalapril regimen). At week 4, a substantial number of patients remained on monotherapy at either the initial dose or double the dose of losartan (52%) or enalapril (72%). The blood pressure curves for each treatment were parallel over time. The enalapril-based regimen elicited a statistically significantly greater reduction in blood pressure than the losartan-based regimen, although the mean differences in the blood pressure response between the two treatment groups was small. Based on sitting diastolic blood pressure < 90 mmHg or a reduction in blood pressure of at least 10 mmHg, 98% of the patients assigned to the losartan regimen and 100% of the patients assigned to the enalapril regimen had a satisfactory response with a regimen of one to three antihypertensive drugs. Headache was the most common adverse experience in both treatment groups (occurring in 22% of patients assigned to the losartan regimen and 20% of patients assigned to the enalapril regimen). CONCLUSIONS: In this study, the losartan-based regimen effectively lowered blood pressure, was generally well tolerated, and was generally similar to the enalapril-based regimen in the treatment of patients with severe hypertension. PMID- 8728307 TI - Cardiac source impedance and the pressure/flow characteristics of an arterial load. PMID- 8728308 TI - Day-night differences in blood pressure and left ventricular mass. PMID- 8728309 TI - Role of cholesterol in regulating apolipoprotein B secretion by the liver. AB - The review examines the evidence that the supply of cholesterol available for incorporation into nascent lipoprotein particles exerts a regulatory influence on apolipoprotein (apo) B secretion by the liver. Support for this hypothesis comes both from in vitro experiments and from recent observations in normal subjects and patients with dyslipidemia associated with familial hypercholesterolemia, obesity, noninsulin dependent diabetes mellitus, growth hormone deficiency and cholesteryl ester storage disease. The findings do not negate a role for triglyceride synthesis in determining apoB secretion in very low density lipoprotein, but the inhibitory effects on the latter process of pharmacological blockade of cholesterol synthesis or esterification suggest that it is conditional upon an adequate supply of cholesteryl ester. PMID- 8728310 TI - Stimulation of the activities of hepatic fatty acid oxidation enzymes by dietary fat rich in alpha-linolenic acid in rats. AB - The activities of hepatic fatty acid oxidation enzymes in rats fed perilla oil rich in alpha-linolenic acid (alpha-18:3) were compared with those fed saturated fats or safflower oil (the mixture of safflower oil and olive oil, 94:8, w/w) containing the same amount of polyunsaturated fatty acids with perilla oil exclusively as linoleic acid (18:2). When the rats were fed the diets containing 15% coconut, safflower, and perilla oils for 1 week, the rate of mitochondrial and peroxisomal oxidation of palmitoyl-CoA (16:0-CoA) in the liver homogenates was the highest in rats fed perilla oil. Among the rats fed the diets containing 15% palm, safflower, and perilla oils for 2 weeks, the rates of mitochondrial and peroxisomal oxidations of 16:0-, 18:2-, and alpha-18:3-CoAs were the highest in rats fed perilla oil, and the rate of oxidation of alpha-18:3-CoA by both pathways was higher than those of other acyl-CoAs in all groups. Dietary perilla oil relative to palm and safflower oils significantly increased the activities of carnitine palmitoyltransferase, acyl-CoA dehydrogenase, acyl-CoA oxidase, and 2,4 dienoyl-CoA reductase. The substrate specificity of carnitine palmitoyltransferase appeared to be responsible for differential rates of the mitochondrial oxidation of acyl-CoAs. The substrate specificity of acyl-CoA oxidase did not account for the preferential peroxisomal oxidation of alpha-18:3 relative to 18:2. The preferential mitochondrial and peroxisomal beta-oxidation of alpha-18:3-CoA relative to 16:0- and 18:2-CoAs was also confirmed in rats fed laboratory chow irrespective of the substrate/albumin ratios in the assay mixture. It was suggested that both substrate specificities and alterations in the activities of the enzymes in beta-oxidation pathway play a significant role in the regulation of the serum lipid concentrations in rats fed a diet rich in alpha-18:3. PMID- 8728311 TI - Lipoprotein lipase-mediated uptake of lipoprotein in human fibroblasts: evidence for an LDL receptor-independent internalization pathway. AB - Lipoprotein lipase (LPL), a key enzyme in lipoprotein triglyceride metabolism, produces a marked increase in the retention and uptake of all classes of lipoproteins by cultured cells. It was previously shown that two different receptors are involved in mediating the LPL effects: heparan sulfate proteoglycans (HSPG) and the low density lipoprotein (LDL) receptor-related protein/alpha 2 macroglobulin receptor (LRP). By immunofluorescence we show here that cell surface-bound LPL displays a pattern that corresponds to the previously described distribution of cell surface HSPG. No evident relation to the distribution of bound activated alpha 2-macroglobulin (alpha 2M*) or to LRP was observed. By immunoelectron microscopy we found that after 30 min at 37 degrees C most of the detected alpha 2M* (70% of the total gold particles) was inside the cells and associated with endosomal vesicles. However, at the same time, 76% of the LPL remained at the cell surface, suggesting that, LPL is internalized by a slow endocytic process. Binding of triglyceride-rich lipoproteins (TRL) or LDL together with LPL led to a spectacular increase in bound lipoproteins, which completely colocalized with LPL. After incubation at 37 degrees C, LPL and 1,1' dioctadecyl-3,3,3,'3'-tetramethylindocarbocyanine (DiI)-TRL formed large clusters on the cell surface. Immunofluorescene and quantitative immunoelectron microscopy provided evidence of co-internalization of LPL and apoE-containing TRL by a slow endocytic process. In the absence of LPL, the fibroblasts rapidly internalized DiI-LDL and showed fluorescence in central, lysosome-like vesicles. In contrast, when LPL was present, internalization of DiI-LDL involved small, widely distributed vesicles. This pattern slowly changed to one consisting of large perinuclear vesicles. LDL receptor-deficient fibroblasts internalized DiI-LDL, either with or without LPL, into small widely distributed vesicles and no central vesicles were seen. Chloroquine-treated normal fibroblasts internalized DiI-LDL in a pattern similar to that of receptor-deficient fibroblasts. Taken together our results suggest an alternative receptor-independent endocytosis pathway for LDL. This pathway is potentiated by LPL and is characterized by a slow uptake involving small vesicles that gradually reach lysosomes. We suggest that, through its interaction with HSPG, LPL provides high capacity binding sites for lipoproteins and a independent internalization pathway. PMID- 8728312 TI - In vivo biosynthesis of retinoic acid from beta-carotene involves and excentric cleavage pathway in ferret intestine. AB - This study was done to determine whether retinoic acid can be produced by excentric cleavage of beta-carotene in vivo. By using an inhibitor of retinaldehyde oxidation, citral, either retinaldehyde or beta-carotene was incorporated in a micellar solution and perfused through the upper portion of small intestine of ferrets. After 2 h perfusion of 1 microM retinaldehyde, retinoic acid rose in portal blood (+3.5 +/- 1.3 nmol/L) and was detected in the intestinal mucosa (30 +/- 2 pmol/g). When citral was added at 2 mM along with retinaldehyde, retinoic acid decreased in the portal blood and retinoic acid was not detected in the intestinal mucosa. With or without the presence of citral (2 mM), the perfusion of beta-carotene (10 microM) during 2 h caused a significant rise of retinoic acid in portal blood (+2.6 +/- 0.6 nmol/L and + 4.1 +/- 0.6 nmol/L, respectively) and in liver; moreover, significant amounts of retinoic acid were detected in the intestinal mucosa (19 +/- 3 pmol/g and 36 +/- pmol/g, respectively. This study demonstrates that after intestinal perfusion of beta carotene in the ferret in vivo, a substantial amount of retinoic acid is formed via an excentric cleavage pathway. PMID- 8728313 TI - Synthesis and secretion of B-100 and A-I apolipoproteins in response to the changes of intracellular cholesteryl ester content in chick liver. AB - We investigated in the chick whether the diet-induced changes of the hepatic content of cholesteryl esters (CE) influence the synthesis and the secretion of apoB- and apoA-I-containing lipoproteins. Control chicks received a low cholesterol diet for 2 (SD-1), 4 (SD-2), or 7 (SD-3) weeks; the chicks in the experimental groups received a cholesterol-rich diet for 2 weeks and were killed at the end of the cholesterol feeding (CH-F), and after 2 (CH-D) or 5 (CH-DD) weeks of a low cholesterol diet. Hepatic CE content in CH-F chicks was 30-fold that observed in controls, but returned to the control level after 5 weeks of cholesterol depletion (CH-DD). The incorporation of 35S-labeled amino acids into cell and medium apoB and apoA-I was measured in liver slices. Intracellular 35S labeled apoB was similar in all groups whereas medium 35S-labeled apoB was 2-fold higher in CH-F than in controls (SD-1). Pulse-chase experiments showed that radioactive apoB secreted by CH-F chicks at 120 min of chase was 2 times that of SD-1 chicks. This increased secretion of apoB was not found in CH-D chicks. In CH F chicks, the intracellular and medium 35S-labeled apoA-I were 2-fold the values found in controls (SD-1); apoA-I production returned to the control level only after 5 weeks of cholesterol depletion (CH-DD). The increased secretion of apoB and apoA-I in CH-F chicks was associated with an increased secretion of very low, intermediate, and low density lipoproteins containing newly synthesized apoB and apoA-I and of high density lipoproteins containing predominantly apoA-I. Thus, in response to hepatic CE accumulation induced by cholesterol feeding, a larger proportion of newly synthesized apoB is driven to the secretory pathway and more apoA-I is synthesized. This promotes an increased secretion of plasma lipoproteins that contribute to the removal of CE from the liver. PMID- 8728314 TI - Apolipoproteins C-III and E in apoB- and non-apoB-containing lipoproteins in two populations at contrasting risk for myocardial infarction: the ECTIM study. Etude Cas Temoins sur 'Infarctus du Myocarde. AB - Apolipoprotein (apo) C-III and apoE are components of two major classes of plasma lipoproteins, i.e., apoB- and non-apoB-containing lipoproteins. To analyze the relationship between the distribution of apoC-III and apoE among lipoproteins and coronary heart disease, we compared the distribution of these two apolipoproteins in survivors of myocardial infarction (MI) and control subjects, within and between populations at contrasting risk for MI. ApoC-III and apoE concentrations were determined in plasma devoid of apoB-containing lipoproteins by immunoprecipitation using a specific anti-apoB antiserum. These assays referred to apoC-III-Lp non-B and apoE-Lp non-B, respectively. By examining the difference with total plasma apoC-III and apoE levels, we calculated apoC-III and apoE in apoB-containing lipoproteins (apoCIII-LpB and apoE-LpB, respectively). These determinations were performed in control subjects and in survivors of MI, all males aged 25 to 64 years. They were recruited in Northern Ireland and France, countries characterized by a large difference in the incidence of coronary heart disease. In univariate analysis, apoCIII-LpB appeared significantly higher and the apoC-III ratio (apoC-III-Lp non-B/apoC-III-LpB) significantly lower in MI survivors than in control subjects in both countries. ApoE-LpB was higher in MI survivors than in control subjects in Northern Ireland but not in France. The two French and Irish control populations differed for apoC-ILL-Lp non-B, apoC-III ratio, and apoE ratio, which were higher in France than in Northern Ireland, and for apoC-III-LpB, apoE, and apoE-LpB, which were lower. Multivariate analysis showed that no parameter involving apoC-III and apoE was more discriminatory than HDL-cholesterol, cholesterol, and triglycerides or apoA-I, apoB, and triglycerides between controls and MI subjects. In contrast, the apoC-III ratio was a better discriminatory parameter between the two control populations than the listed parameters. The differences between the two control populations are of particular interest because they are not biased by the presence of disease or the large difference of the incidence in coronary heart disease between the two countries. It is suggested that the distribution of apoC-III among lipoproteins may play a role in the different susceptibility of the two populations to the atherogenic process. PMID- 8728315 TI - Interaction of a recombinant form of apolipoprotein[a] with human fibroblasts and with the human hepatoma cell line HepG2. AB - We have studied the binding, uptake, and degradation of a recombinant form of apolipoprotein[a] (r-apo[a]) using a cultured cell model. In HepG2 cells and in human fibroblasts, r-apo[a] complexed with low density lipoprotein(LDL) is bound and internalized via high affinity (Kd = 10 nM) receptors; in both cell types, low affinity (Kd = 200-300 nM) sites also mediate free apo[a] uptake. Using competition studies, we found that the high affinity binding component corresponds to the LDL receptor. Involvement of the LDL receptor in r-apo[a] uptake by fibroblasts was confirmed using fibroblasts derived from an individual homozygous for familial hypercholesterolemia; in contrast to normal fibroblasts, these cells lacked the high affinity r-apo[a] binding component. Cell association of 125I-labeled r-apo[a] was increased and decreased concomitantly with the up- and down-regulation of the LDL receptor in response to a number of compounds. The addition of alpha 2-macroglobulin as well as treatment with heparinase, chondroitinase ABC, and sodium chlorate did not decrease total specific binding of r-apo[a], suggesting that neither the low density lipoprotein receptor-related protein nor cell surface proteoglycans are involved in r-apo[a] clearance. The low affinity binding component present in both fibroblasts and HepG2 cells likely corresponds to the plasminogen receptor, as binding of r-apo[a] to these sites was specifically decreased by the addition of plasminogen or the lysine analogue epsilon-aminocaproic acid, but not by the addition of tissue-type plasminogen activator. Heparin abolished uptake of r-apo[a] by the LDL receptor component only; this indicates that apo[a] must be associated with LDL to be cleared by this receptor. In contrast, free apo[a] can be effectively cleared by the plasminogen receptor which may represent a significant route of clearance for free apo[a] in vivo. PMID- 8728316 TI - Cholesterol homeostasis is modulated by amphiphiles at transcriptional and post transcriptional loci. AB - A variety of amphiphiles inhibit plasma membrane cholesterol esterification and induce 3-hydroxy-3-methylglutaryl-coenzyme A reductase accumulation in cultured cells; among these are steroids, hydrophobic amines, phenothiazines, ionophores, colchicine, and lysophosphatides. It has been proposed that these amphiphiles signal a sterol deficiency to regulatory sites by blocking the movement of plasma membrane cholesterol into the cell (Lange, Y., and Steck, T. L. 1994. J. Biol. Chem. 269: 29371-29374). If this were the case, these agents also should enhance transcription of sterol responsive genes and stabilize 3-hydroxy-3-methylglutaryl coenzyme A reductase. As a test of this hypothesis, the effect of the amphiphiles on such transcriptional and post-transcriptional events was assessed. A mouse embryo cell line was transfected with a construct containing the promoter for the human low density lipoprotein receptor upstream of the DNA sequence coding for chloramphenicol acyltransferase (CAT). Incubation of these cells for 7-18 h with the aforementioned agents caused the level of expression of the promoter/CAT construct to increase 2- to 9-fold. We showed further that the amphiphiles stimulated 3-hydroxy-3 methylglutaryl-coenzyme A reductase activity by increasing gene transcription as well as by decreasing degradation of the enzyme. These are the predicted homeostatic responses to cell cholesterol deficiency. These findings support the hypothesis that certain amphiphiles falsely signal a cholesterol deficiency to the intracellular sites regulating cholesterol homeostasis. PMID- 8728317 TI - Formation of delta 22-bile acids in rats is not gender specific and occurs in the peroxisome. AB - We recently demonstrated that the formation of delta 22-bile acids is a quantitatively major pathway for normal bile acid synthesis in the adult male Sprague-Dawley rat. This pathway is specific for 7 beta-hydroxy bile acids and, when ursodeoxycholic acid is administered, delta 22-ursodeoxycholic acid appears as a major metabolite in the liver tissue, bile, intestinal contents, and plasma. The aims of this study were, therefore, to determine whether this metabolic pathway was gender specific, and to establish that the peroxisome is a site of formation of delta 22-bile acids. Bile acids were determined by gas chromatography-mass spectrometry in liver tissue, jejunum, and plasma of adult female rats and in animals fed a diet containing 0.4% and 1% ursodeoxycholic acid. Bile acid metabolism in female rats was found to be similar to that of male rats, and delta 22-beta-muricholic acid, rather than beta-muricholate, was likewise confirmed as the major muricholic acid synthesized. Ursodeoxycholic acid administration resulted in the appearance of delta 22-ursodeoxycholic acid as a major metabolite. When adult male Sprague-Dawley rats were treated with clofibrate, a drug that induces peroxisomal proliferation, liver weight increased 40-60% and total bile acid synthesis decreased markedly, but the relative composition of individual bile acids was unchanged. When ursodeoxycholic acid was added to the diet, the proportion of delta 22-bile acids relative to the corresponding saturated analogues increased significantly compared with untreated rats, indicating that clofibrate had "amplified" the pathway for formation of delta 22-bile acids. When UDCA was incubated in vitro with a peroxisomal-enriched fraction from normal adult male rat liver, delta 22-ursodeoxycholic acid was formed in proportions comparable to that observed in vivo when this bile acid was given orally. These studies establish that the pathway for the formation of delta 22-bile acids is not gender specific and mainly occurs in hepatic peroxisomes. PMID- 8728318 TI - Glycosylation structure and enzyme activity of lecithin:cholesterol acyltransferase from human plasma, HepG2 cells, and baculoviral and Chinese hamster ovary cell expression systems. AB - The glycosylation state of lecithin:cholesterol acyltransferase (LCAT) may be important in determining its enzymatic activity. We compared glycosylation structure, enzyme kinetics, and phosphatidylcholine (PC) acyl specificity of human LCAT from four sources: human plasma (pLCAT), media from HepG2 cells (HepG2 LCAT), media from SF21 cells infected with a recombinant baculovirus (bLCAT) and media from stably transfected Chinese hamster ovary (CHO) cells (CHO LCAT). bLCAT was underglycosylated (molecular weight approximately 50 kDa) and resistant to digestion by N-glycanase F, endoglycosidase F, and neuraminidase. CHO and HepG2 LCAT were overglycosylated (approximately 68 kDa and approximately 70-75 kDa) compared to pLCAT (approximately 65 kDa). CHO LCAT, like pLCAT, was sensitive to N-glycanase F and neuraminidase but not to endoglycosidase F. HepG2 LCAT demonstrated resistance to N-glycanase F and endoglycosidase F. Apparent Km values for all four enzymes were similar (1.4-9.2 microM cholesterol) for recombinant high density lipoproteins (rHDL) containing sn-1 16:0, sn-2 18:1 PC (POPC). Apparent Vmax values (nmol cholesteryl ester formed/h per micrograms) were 52.6 for pLCAT, 48.6 for CHO LCAT, 15.3 for bLCAT, and 8.3 for HepG2 LCAT. Changes in PC acyl specificity in the presence and absence of cholesterol were characterized by comparing the ratio of LCAT activity on rHDL containing sn-1 16:0, sn-2 20:4 PC (PAPC) or POPC (PAPC/POPC activity ratio). The ratios for pLCAT, bLCAT, CHO LCAT, and HepG2 LCAT activity were 0.63, 0.49, 0.56, and 0.51 with cholesterol and 0.34, 0.29, 0.36, and 0.99 without cholesterol, respectively. We conclude that LCAT source influences glycosylation structure, which affects the apparent Vmax for cholesteryl ester formation with only minor changes in apparent Km or acyl substrate specificity. PMID- 8728319 TI - Adsorption of mixtures of bile salt taurine conjugates to lecithin-cholesterol membranes: implications for bile salt toxicity and cytoprotection. AB - Tauroursodeoxycholate (TUDC), a relatively hydrophilic bile salt, reduces disruption of cholesterol-rich membranes by more hydrophobic bile salts such as taurocholate (TC), taurochenodeoxycholate (TCDC), or taurodeoxycholate (TDC). We examined the interactions of these bile salts in adsorption to large unilamellar vesicles to determine whether TUDC may stabilize membranes by preventing adsorption of more toxic bile salts. Fractional adsorption was quantified by rapid ultrafiltration. Adsorption coefficient Ai was defined for each bile salt i as ([bound i]/[free i])/[lecithin]. Affinity of different bile salts for lecithin vesicles varied with their relative hydrophobicity, increasing in the order TUDC < TC << TCDC < or = TDC. Ai of each bile salt fell with its accumulation on membranes, reaching a minimum at bound bile salt/lecithin mole ratio (B:L) between 0.05 and 0.1, then increasing with formation of higher-affinity mixed micelles. Inclusion of cholesterol in vesicles reduced Ai of all bile salts. In heterologous binding studies at submicellar concentrations, Ai of each bile salt varied with total B:L but was independent of the specific bile salts present on the membrane. Addition of TUDC to TDC reduced binding of TDC to membranes only slightly and lowered the threshold TDC concentration associated with transition to mixed micelles. However, above this threshold, TUDC markedly altered the adsorption of TDC to lecithin-containing phases. We conclude that TUDC does not directly stabilize membranes; rather, reduced permeabilization and dissolution of cholesterol-rich membranes after addition of TUDC to TDC may result from effects on the formation and structure of simple and mixed micelles. PMID- 8728320 TI - Modulation of macrophage scavenger receptor transport by protein phosphorylation. AB - The identification of three highly conserved phosphorylation sites in the cytoplasmic domain of each of the monomeric subunits of the macrophage scavenger receptor suggests that protein phosphorylation may regulate this receptor pathway. To investigate this, mouse peritoneal macrophages were pretreated with either the protein phosphatase inhibitor okadaic acid or the protein kinase inhibitor staurosporine to modulate cellular protein phosphorylation and their effects on the metabolism of acetyl-LDL were measured. Both okadaic acid and staurosporine inhibited the degradation of acetyl-low density lipoprotein (LDL) without affecting cellular lactic dehydrogenase (LDH) levels. The inhibition by okadaic acid was due to a 70% decrease in acetyl-LDL binding whereas post receptor processing was minimally affected. Calyculin A, another serine/threonine phosphatase inhibitor, also reduced acetyl-LDL binding, whereas lithium chloride, an inositol phosphatase inhibitor, did not. Okadaic acid did not decrease steady state receptor mRNA levels nor decrease the number of total cellular receptors, consistent with a posttranslational mechanism of action. Interestingly, protease sensitivity studies showed that the receptors were still located on the cell surface. These studies suggest that okadaic acid inhibits acetyl-LDL binding by causing the redistribution of surface receptors into a sequestered compartment or inactivating the receptors. In contrast, staurosporine produced a paradoxical increase in receptor expression (30%) but slowed post-receptor processing (2.3 fold decrease). The latter was due to an inhibition of ligand internalization (2.9-fold decrease) via a protein kinase C-independent mechanism. Macrophage pinocytosis was also slowed by staurosporine (38% decrease); however, this does not appear to account for the inhibition of scavenger receptor internalization. Direct receptor phosphorylation was also slowed by staurosporine (38% decrease); however, this does not appear to account for the inhibition of scavenger receptor internalization. Direct receptor phosphorylation was also investigated and it was established that the receptor can be phosphorylated; however, changes in receptor function did not correlate with changes in the degree of receptor phosphorylation. Together these studies demonstrate that changes in cellular protein phosphorylation affect the expression, surface transport, and internalization of the macrophage scavenger receptor and suggest that the regulated phosphorylation/dephosphorylation of cellular proteins may be an important biochemical mechanism that controls normal processing of ligands by this receptor pathway. PMID- 8728321 TI - Rat hepatoma L35 cells, a liver-differentiated cell line, display resistance to bile acid repression of cholesterol 7 alpha-hydroxylase. AB - A stable hepatoma cell line (L35 cells) showing an activation of the cholesterol 7 alpha-hydroxylase gene (CYP7) that had been silent in the parental hepatoma cell line (H35 cells) was used to examine the influence of bile acids on its gene expression and activity. L35 cells were found to concentrate taurocholate from the culture medium, without any significant effect on the expression of 7 alpha hydroxylase. At physiologic levels (up to 100 microM), CYP7 mRNA expression was not repressed by any bile acid. At supra-physiologic levels (1 mM), the more hydrophobic dihydroxy bile acids, taurodeoxycholate and taurochenodeoxycholate, decreased CYP7 mRNA without decreasing the relative abundance of beta-actin mRNA. Similar results were obtained by culturing cells with sodium dodecylsulfate (50 microM). The medium of L35 cells treated with either taurochenodeoxycholate (1 mM), taurodeoxycholate (1 mM), or sodium dodecylsulfate (50 microM) contained significantly greater activities of two cytosolic enzymes, lactate dehydrogenase and phosphoglucose isomerase, indicating a cytotoxic response. Activation of protein kinase C by phorbol esters decreased the expression of 7 alpha hydroxylase mRNA without evidence of cytotoxicity; therefore, the inability of L35 cells to show bile acid repression cannot be ascribed to a lack of an effect by this secondary messenger system. In addition, insulin decreased and dexamethasone increased 7 alpha-hydroxylase mRNA without increasing the release of the cytoplasmic enzyme markers. The combined data suggest that L35 cells are resistant to repression of CYP7 gene expression by bile acids, but display physiologic expression to hormones and protein kinase C activation. PMID- 8728322 TI - Expression and secretion of rabbit plasma cholesteryl ester transfer protein by Pichia pastoris. AB - The rabbit cholesteryl ester transfer protein (CETP) was expressed in the methylotrophic yeast Pichia pastoris by introducing the CETP cDNA under the control of the methanol-inducible alcohol oxidase promoter. The cDNA was cloned from in vitro amplified cDNA of rabbit liver mRNA. The nucleotide sequence of the cloned cDNA differed slightly from the previously published sequence that changed the amino acid sequence in six residues. Interestingly, five of these replacements are identical to the corresponding residues in human CEPT. In addition, the encoded mature N-terminal sequence was changed from Cys- to Arg-Glu Phe- to link the CETP sequence to the yeast acid phosphatase signal peptide. The culture medium of the transformed cells induced with 1% methanol contained both cholesteryl ester and triglyceride transfer activity comparable to that of rabbit plasma. Like rabbit plasma, the lipid transfer activity in the medium could be inhibited by monoclonal antibodies that block CE/TG transfer or TG transfer alone. Immunoblot analysis of M(r) = 80 K and minor species of M(r) = 60-100 K. In spite of these differences, the specific transfer activity of the recombinant CETP was indistinguishable from that of rabbit plasma CETP of M(r) = 74 K. N Glycosidase F treatment converted both the recombinant and plasma CETP to a single species of M(r) = 55 K. Both the plasma and recombinant CETP lost their activity after removal of N-linked carbohydrate and sialic acid. A single 55 K component was found in the cell-lysates. The intracellular form of the recombinant CETP was not modified by N-glycosidase F treatment. In conclusion, the recombinant CETP is synthesized as an inactive polypeptide that is processed and secreted as a functional glycoprotein. In addition, the N-terminal Cys residue of the plasma CETP is not required for its activity. PMID- 8728324 TI - A novel mutation in the cytochrome P450(27) (CYP27) gene caused cerebrotendinous xanthomatosis in a Japanese family. AB - Cerebrotendinous xanthomatosis (CTX) is an autosomal recessive lipid storage disease caused by mutations in the cytochrome P450(27) (CYP27) gene. This disease is characterized by the accumulation of a bile alcohol, cholestanol, in diverse tissues. Accumulation in the central nervous system leads to neurological dysfunction including dementia, spinal cord paresis, and cerebellar ataxia. Accumulation in other tissues causes tendon xanthomas, premature atherosclerosis, and cataracts. In a Japanese family with CTX, we identified two points mutations in the CYP27 gene at different sites. One is a novel transversion, which substitutes G for C at Pro 368 (CCC) to Arg (CGC). The other is a transition, which substitutes A for G at Arg441 (CGG) to Gln (CAG), this being the same mutation that Kim et al. reported (1994. J. Lipid Res. 35: 1031 - 1039). Allele specific polymerase chain reaction analysis indicated that the father and mother of this family, who themselves had no clinical manifestations of CTX, had the former and latter mutations heterozygously, respectively. On the other hand, the patients each had both mutations heterozygously. These results are highly suggestive, but not conclusive, that the newly identified transversion in the CYP27 gene accounts for the sterol 27-hydroxylase (EC 1.14.13.15) deficiency in these patients. PMID- 8728323 TI - Complete mapping of crystallization pathways during cholesterol precipitation from model bile: influence of physical-chemical variables of pathophysiologic relevance and identification of a stable liquid crystalline state in cold, dilute and hydrophilic bile salt-containing systems. AB - Using complementary physical-chemical techniques we defined five different crystallization pathways as functions of time (30 days) and increasing lecithin (egg yolk) content in pathophysiologically relevant model biles super-saturated (cholesterol saturation indices, 1.2 - 2.7) by dilution of approximately equal to 29 g/dl bile salt-lecithin-cholesterol micellar solutions. As evidenced by quasi elastic light-scattering spectroscopy, supersaturation was heralded by the appearance of unilamellar vesicles. With the lowest lecithin contents, arc-like crystals with habit and density (d 1.030 g/mL) consistent with anhydrous cholesterol appeared first and evolved via helical and tubular crystals to form plate-like cholesterol monohydrate crystals (d 1.045 g/mL). With higher lecithin fractions, cholesterol monohydrate crystals appeared earlier than arc and other transitional crystals. With typical physiological lecithin contents, early liquid crystals (d 1.020 g/mL) were followed by cholesterol monohydrate crystals and subsequent appearances of arc and other intermediate crystals. With higher lecithin contents, liquid crystals were followed by cholesterol monohydrate crystals only, and at the highest lecithin mole fractions, liquid crystals appeared that did not generate solid crystals. Added calcium increased solid crystal number in proportion to its concentration (5 - 20 mM) but did not influence appearance times, crystallization pathways, or micellar cholesterol solubilities. Decreases in temperature (37 degrees --> 4 degrees C), total lipid concentration (7.3 --> 2.4 g/dL), and bile salt hydrophobicity (3 alpha, 12 alpha --> 3 alpha, 7 alpha, 12 alpha --> 3 alpha, 7 beta hydroxylated taurine conjugates) progressively shifted all crystallization pathways to lower lecithin contents, retarded crystallization, and decreased micellar cholesterol solubilities. The lecithin content of mother biles decreased markedly during crystallization especially where liquid crystals were a coexisting phase at equilibrium. This systematic study provides a framework for understanding cholesterol crystallization in human and animal biles and for examining factors that influence the kinetics of phase separation. PMID- 8728325 TI - Alterations in plasma lipoproteins and apolipoproteins before the age of 40 in heterozygotes for lipoprotein lipase deficiency. AB - We have assessed the expression of heterozygosity for lipoprotein lipase (LPL) deficiency by studying a single large French Canadian family comprising 92 persons including 21 carriers of the catalytically defective P207L mutation. Phenotypic changes distinguishing heterozygotes from controls were seen early, before the age of 40 and often before 20 years of age. In the total cohort these changes included an elevation in the mean very low density (VLDL) and intermediate density lipoprotein (IDL) triglyceride (+69%; P = 0.01 and +40%; P = 0.001) and cholesterol (+51%; P = 0.03 and +67%; P = 0.007) and apoB levels but decreased HDL2 and HDL3 cholesterol, (-32%; P = 0.01 and -15%; P = 0.002 respectively). While the lipid compositions of VLDL and IDL were similar between heterozygotes and controls, the low density (LDL) and high density lipoproteins (HDL) of carriers were triglyceride enriched. Heterozygotes also had a markedly lower apoC-III ratio (apoC-III in supernatant/apoC-III in heparin precipitate) (1.46 vs. 3.86 P = 1 x 10(-4)) indicating a substantial enrichment of VLDL and IDL with apoC-III and depletion of HDL apoC-III supporting this ratio as an effective index for efficiency of lipolysis. LpA-I was markedly reduced (0.34 vs. 0.43 P = 1 x 10(-5)) showing that levels of this particle are partly dependent on LPL catalytic activity. Heterozygotes manifest from an early age with a markedly reduced HDL, LpA-I, apoC-III ratio and an increased TC/HDLc ratio which would predict a relatively increased risk of premature coronary artery disease, compared to their normal siblings. PMID- 8728326 TI - Homozygosity for two point mutations in the lipoprotein lipase (LPL) gene in a patient with familial LPL deficiency: LPL(Asp9-->Asn, Tyr262-->His). AB - Familial lipoprotein lipase (LPL) deficiency is an inherited disorder of lipoprotein metabolism characterized by hypertriglyceridemia and recurrent episodes of abdominal pain and pancreatitis. We have studied the genetic basis of LPL deficiency in a 62-year-old black male with undetectable pre- and post heparin plasma LPL mass and activity, DNA sequence analysis of the patient's LPL cDNA and gene as well as digestion with Bcl I and Asu I revealed that the proband is a homozygote for two separate gene defects. One mutation changed a G to an A, resulting in the conversion of amino acid 9 of the mature protein, aspartic acid (GAC), to asparagine (AAC). The second substitution, a C for a T, replaced tyrosine (TAC) at residue 262 with histidine (CAC). Northern blot analysis of monocyte-derived macrophage RNA demonstrated the presence of LPL mRNA of approximately normal size and quantity when compared to control. Expression of both mutations separately (pCMV-9 and pCMV-262) or in combination (pCMV-9+262) in human embryonal kidney-293 cells demonstrated that LPL-9 had approximately 80% the specific activity of wild type LPL, but LPL-262 and LPL-9+262 had no enzymic activity, thus establishing the functional significance of the LPL-262 defect. Despite an absolute deficiency of LPL mass and activity demonstrated by analysis of patient post-heparin plasma, in vitro expression of both LPL mutants was normal, suggesting that the absence of LPL in patient post-heparin plasma was a result of altered in vivo processing. Analysis of the heparin binding properties of the mutant enzymes by heparin-Sepharose affinity chromatography indicated that most of the LPL-262 mass was present in an inactive peak, which like the normal LPL monomer, eluted at 0.8 M NaCl. Thus, the Tyr262 --> His mutation may alter the stability of the LPL dimer, leading to the formation of inactive LPL-262 monomer which exhibits reduced heparin affinity. Based on these results, we propose that, in vivo, enhanced formation of LPL-9+262 monomer leads to abnormal binding of the mutant lipase to endothelial glycosaminoglycans ultimately resulting in enhanced catabolism of the mutant enzyme and lower enzyme mass in post-heparin plasma. PMID- 8728327 TI - Partial deletion of the gene encoding sterol 27-hydroxylase in a subject with cerebrotendinous xanthomatosis. AB - An Italian subject with cerebrotendinous xanthomatosis (CTX) was found to have a partial deletion of the gene encoding the enzyme sterol 27-hydroxylase (CYP27 gene). Southern blot analysis revealed that this deletion (approximately 2 kb) spans from intron 6 to the 3' flanking (3'FLK) region, eliminating exons 7-9, the last three exons of CYP27 gene. No sterol 27-hydroxylase mRNA was detected in proband cells, either by Northern blot analysis or by reverse transcription polymerase chain reaction (PCR). This suggests that the mutant mRNA devoid of the exon encoding the whole untranslated sequence (exon 9) might be rapidly degraded in the cytoplasm. We used inverse PCR to obtain a partial sequence of the 3'FLK region of the normal CYP27 gene; this allowed us to define the mechanism underlying the deletion. The established sequence was used to design suitable primers to perform step-wise sequences of a 1.7 kb segment of the 3'FLK region of the normal gene and of the deletion joint in the CTX patient. The analysis of the sequence data indicate that the deletion might result from a complex mechanism involving two intragenic recombinations between a) two 14 nucleotide complementary sequences, one in intron 6 and the other in the 3'FLK region: and b) AT-rich complementary sequences of the 3'FLK region, and a slipped mispairing between two 6 nucleotide direct repeats, one in intron 6 and the other in the 3'FLK region. Such repeats are brought close to each other by the formation of the stem-loops induced by the two intragenic recombinations. This is the first example of CTX caused by a rearrangement of CYP27 gene. PMID- 8728328 TI - High level secretion of wild-type and mutant forms of human proapoA-I using baculovirus-mediated Sf-9 cell expression. AB - To facilitate the investigation of apoA-I structure:function relationships as they relate to LCAT activation and lipid binding, we have developed an apoA-I baculoviral expression and purification system that yields milligram quantities of wild-type or mutant proapoA-I. Baculovirus-infected Sf-9 cells, grown in suspension, were found to secrete high levels of human wild-type (40-50 mg/l) or mutant apoA-I protein (1-38 mg/l), which was determined to be > 95% pure following a two-step purification procedure. In the case of wild-type apoA-I, ELISA showed that approximately 13-18% of the total protein secreted into the culture medium was apoA-I. To isolate pure protein from culture medium, 72 h post infection medium was subjected to preparative reverse phase high performance liquid chromatography (HPLC), followed by DEAE ion-exchange chromatography. Purity and molecular size determination of wild-type proapoA-I protein was verified by SDS polyacrylamide gel electrophoresis, electrospray mass spectrometry, and N-terminal sequencing. In addition, recombinant discoidal apoA I:phospholipid complexes prepared from wild-type or plasma apoA-I showed similar particle size and LCAT activation properties. To fully characterize the utility of this expression system, the expression levels of various mutant apoA-I proteins were compared to wild-type. Despite a lower production level seen with selected apoA-I mutants, milligram quantities of these purified mutant proteins were also obtained. In summary, we show that baculovirus-derived wild-type proapoA-I shows properties similar to plasma apoA-I relative to recombinant HDL formation, LCAT reactivity, and alpha-helical content. In addition, we show that a variety of mutant forms of human proapoA-I can be expressed and purified in abundant quantity from baculoviral-infected Sf-9 cells. PMID- 8728329 TI - High performance liquid chromatography of long-chain acylcarnitine and phospholipids in fatty acid turnover studies. AB - In this paper we describe a rapid, isocratic high performance liquid chromatography (HPLC) method for the study of radioactive fatty acid incorporation into complex lipids of human erythrocytes, which allows the simultaneous separation of the major phospholipid classes and long-chain acylcarnitines. The lipid extract of erythrocytes pulsed with radioactive fatty acids was injected into an HPLC system equipped with a silica column. The individual components eluted were monitored by ultraviolet absorption and radioactive emission. With respect to the UV profile, the radioactive profile showed an additional peak between phosphatidyl-choline and phosphatidylethanolamine, which was identified as long-chain acylcarnitine by different experimental approaches. The radioactivity recovered in the long-chain acylcarnitines contains essential information enabling definition of acyl trafficking in red cells. PMID- 8728341 TI - Clinical pharmacology: present status and projections for the future. PMID- 8728342 TI - Human pharmacokinetics and tolerability of L-365,260, a novel cholecystokinin-B antagonist. AB - A study was conducted to examine the tolerability and pharmacokinetics of single and multiple oral doses of L-365,260, a novel antagonist for type B cholecystokinin (CCK) receptors and to quantify effects of selective blockade of type B CCK receptors through treatment with L-365,260 on measures of anxiety, hunger, and cognitive performance. Healthy volunteers were given single oral doses of up to 50 mg of L-365,260 and multiple oral doses of up to 25 mg every 6 hours for 10 days. Plasma concentrations of L-365,260 were quantified by means of high-performance liquid chromatography. Anxiety and hunger were assessed by visual analog scale and the Spielberger State Anxiety Index. Cognitive testing was used to evaluate attention level and short-term memory. L-365,260 was rapidly absorbed and a biphasic pattern of elimination was demonstrated with a terminal half-life (t1/2) of 8 to 12 hours. The mean (n = 6) values for peak plasma concentration (C(max)) and time to peak concentration (t(max)) of L-365,260 were 503 ng/mL and 1.25 hours, respectively, after a single 50-mg oral dose. Accumulation of L-365,260 plasma concentrations was seen after the prescribed multiple-dose regimens. Steady state was achieved after 3 days of oral administration. L-365,260 had an acceptable tolerability profile after oral administration. No changes in measures of anxiety, hunger, or short-term memory were observed at doses of L-365,260 shown to have antagonist activity at the CCK B receptor. PMID- 8728343 TI - Nonparametric expectation maximization population modeling of ganciclovir. AB - The use of the nonparametric expectation maximization (NPEM2) program to estimate pharmacokinetic parameters of ganciclovir in a group of patients with human immunodeficiency virus (HIV) and cytomegalovirus (CMV) infection was evaluated. A 10-point data set per patient obtained over 8 hours was analyzed. Mean pharmacokinetic parameters obtained included rate constant from the central to the peripheral compartment (KCP,3.1 hr-1), rate constant from the peripheral to the central compartment (KPC, 0.824 hr-1), slope of the volume of distribution to body weight (VS, 0.246 L/kg), and slope of clearance to creatinine clearance (Cl(cr)) and body weight (CLS,0.222L/hr/kg/100 mL/ min Cl(cr). Use of NPEM2 led to identification of a subset of patients with CMV retinitis who had a more rapid clearance of ganciclovir of 0.51 to 0.54 L/hr/kg/100 mL/min Cl(cr). Use of smaller, optimally timed samples of five, four, and three data points per patient produced mean pharmacokinetic parameter results consistent with the full ten point data set. When Bayesian-derived parameter estimates using a five-point data set were compared with a traditional, nonlinear, least-square analysis of the entire ten-point data set, estimates of clearance were determined to be relatively unbiased and precise. The ability of NPEM2 to estimate pharmacokinetic parameters and to determine the population distribution of the parameters was demonstrated. By using points in the analysis chosen by D-optimal design theory, NPEM2 was able to give consistent parameter estimates with as few as three data points. Determination of the distribution appeared to have been dependent on the time points used, however. The approach of MAP-Bayesian analysis to derive patient-specific estimates using optimal samples and prior estimates from a previous population pharmacokinetic analysis for inclusion in subsequent pharmacodynamic analyses of drug exposure (area under the concentration-time curve) may enable development of exposure-response and exposure-toxicity relationships. PMID- 8728344 TI - Pharmacokinetic/pharmacodynamic modeling of cortisol suppression after oral administration of fluocortolone. AB - Fluocortolone is a potent corticosteroid used orally for the treatment of rheumatic diseases, asthma, and immunosuppression. A clinical study of nine healthy volunteers was conducted to determine the pharmacokinetics and cortisol suppression after administration of single oral doses of 20 mg, 50 mg, and 100 mg of fluocortolone. Blood samples were collected at 8:00 AM, 12:00 PM, and 4:00 PM on the day before treatment, and 0, 0.5, 1, 2, 3, 4, 6, 8, 12, 24, 28, 32, 48, 52, and 56 hours after administration of the drug. Concentrations of fluocortolone and cortisol were measured in plasma by a reversed phase high performance liquid chromatography system. Cortisol suppression was chosen as the pharmacodynamic parameter. Total concentrations were converted into unbound concentrations using a two-protein, one-ligand equation. The unbound concentrations were fitted using a one-compartment body model equation with first order absorption. A linear release-rate model was used to characterize the cortisol data. The data were fitted using a common E50 value of 0.95 +/- 0.22 ng/mL for the mean data. The value of E50 was in close agreement with the prediction based on relative glucocorticoid receptor affinity. PMID- 8728345 TI - Pharmacokinetics and bioavailability of mycophenolate mofetil in healthy subjects after single-dose oral and intravenous administration. AB - A randomized, crossover study of 12 healthy volunteers was conducted with single, 1.5-g doses of mycophenolate mofetil (MMF), a prodrug of mycophenolic acid (MPA), after oral and intravenous administration. During the intravenous infusion, phase systemic plasma clearance of MMF was approximately 10 L/min and the half-life (t1/2) was a few minutes. After oral administration, however, plasma MMF was below quantitation limits at all times. The plasma MPA profile of oral MMF showed a sharp peak at approximately 1 hour and a secondary peak at 8 to 12 hours. Mean apparent plasma t1/2 of MPA was similar for both routes (approximately 17 hours). The area under the concentration-time curve (AUC) from time 0 to 24 hours was statistically higher for intravenous than for oral administration, but total AUC showed statistical equivalence (80-120 rule), with mean bioavailability of MPA from oral administration of MMF estimated as 94.1% relative to the intravenous route. Total plasma AUC of mycophenolic acid glucuronide (MPAG), the sole metabolite of MPA, was four- to five-fold higher than MPA. Total 48-hour MPAG recovery in urine was statistically equivalent for the two routes and represented a mean of 70% of administered drug; corresponding MPA recovery was less than 1%. Renal clearance (ClR) values required transport mechanisms for MPAG, but not for MPA. The ClR of MPAG was statistically higher after intravenous administration than oral administration. MMF administered orally undergoes rapid, complete absorption and essentially complete presystemic deesterification. There was presystemic removal of MPA, but enterohepatic circulation compensated for the first pass loss. Renal metabolism of MPA also may have occurred. PMID- 8728346 TI - Effect of time of administration on the pharmacokinetics and tolerance of doxazosin in healthy male volunteers. AB - A randomized, open-label, two-way crossover study of 24 normotensive, healthy male volunteers with nocturia was conducted to compare morning and evening administration of doxazosin in terms of pharmacokinetics and tolerance. In both the morning and evening phases, participants received doxazosin 1 mg once daily for 10 days, followed by 2 mg once daily for 5 days. Pharmacokinetic data were evaluated from blood samples serially collected for 72 hours after drug administration on the last day of each phase. Vital signs and adverse events were recorded throughout the study. Mean peak plasma concentrations (C(max)) were 16.98 and 15.76 ng/mL after morning and evening administration, respectively. Corresponding mean values of area under the plasma concentration-time curve (AUC0 24) were 227.90 and 253.66 ng.hr/mL, respectively. Statistical analysis of the log-transformed values for C(max) and AUC0-24 indicated that morning and evening administration of doxazosin were bioequivalent. There were no statistically or clinically significant differences between phases for mean apparent half-life (t1/2) or total body clearance. There were no clinically relevant differences in blood pressure or in pulse rate between phases, and no occurrences of orthostatic hypotension. The incidence of adverse experiences during morning and evening administration was similar. Morning and evening administration of doxazosin are equivalent and have similar tolerance profiles. PMID- 8728347 TI - Pharmacokinetics of oral mycophenolate mofetil in volunteer subjects with varying degrees of hepatic oxidative impairment. AB - Eighteen patients with compensated alcoholic cirrhosis participated in a single dose pharmacokinetic study of oral mycophenolate mofetil (MMF). Participants were divided into groups of 6 patients each with mild, moderate, or severe hepatic oxidative impairment as defined by the aminopyrine breath test (APBT). Clinically, hepatic disease was of mild or moderate severity. Six healthy volunteers were included as control subjects. Plasma and urine samples were collected over 96 hours and assayed for the active metabolite mycophenolic aced (MPA) and the glucuronide conjugate, MPAG. Plasma protein binding of MPA also was determined in 6 unrelated patients with cirrhosis. Cirrhosis did not grossly affect plasma pharmacokinetics or plasma binding of MPA. Maximum plasma concentrations (C(max)) and area under the curve (AUC) of MPA and MPAG consistently decreased, increased, and then decreased as oxidative impairment declined from normal to severe. Patients with cirrhosis had comparable or greater recovery of administered drug substance in urine than controls, showing that cirrhosis did not affect the extent of MMF absorption. Urine clearance of MPAG was two times higher in the group with severe impairment than in the other groups. Creatinine clearance was similar in all groups. These results suggest progressive impairment of hepatic glucuronidation of MPA and induction of renal glucuronidation in patients with severe hepatic oxidative impairment. PMID- 8728349 TI - Lack of effect of orlistat on the bioavailability of a single dose of nifedipine extended-release tablets (Procardia XL) in healthy volunteers. AB - Orlistat, a lipase inhibitor, reduces dietary fat absorption, and thus could potentially alter the absorption of some concomitantly administered drugs, such as the nifedipine gastrointestinal therapeutic system (GITS). To assess the effect of orlistat on the bioavailability of nifedipine GITS, a third party blind, placebo-controlled, randomized, two-way crossover study was performed in 18 healthy volunteers. Each participant received single 60-mg oral doses of nifedipine GITS (Procardia XL; Pfizer Labs, New York, NY) on the fourth day of treatment with 120 mg of orlistat or placebo three times a day for 6 days. The two treatments were separated by a washout period of at least 1 week. Serial blood samples were collected before and at appropriate intervals after each nifedipine dose to determine plasma concentrations of nifedipine. The 90% confidence intervals for the ratio of geometric least-square means for maximum concentration (C(max)) and area under the concentration-time curve (AUCo-t) and for the difference of arithmetic least-square means for time to maximum concentration (t(max)) indicate that the bioavailability of nifedipine was not altered by treatment with orlistat. Therapeutic doses of 120 mg of orlistat three times daily do not significantly alter the bioavailability of a single 60-mg oral dose of nifedipine GITS in healthy volunteers. PMID- 8728348 TI - Grapefruit juice alters the systemic bioavailability and cardiac repolarization of terfenadine in poor metabolizers of terfenadine. AB - A prospective cohort study was conducted to examine the effects of double strength grapefruit juice on the pharmacokinetics and electrocardiographic repolarization pharmacodynamics of terfenadine in poor metabolizers of terfenadine. Six healthy volunteers who were previously found to be poor metabolizers of terfenadine were studied, with each participant serving as his or her own control. In phase I of the study, terfenadine was given to participants at recommended dosages until steady state was achieved (60 mg twice daily for 7 days). In phase II, participants began receiving concomitant twice-daily, double strength servings of grapefruit juice for 7 days. Serial pharmacokinetic and pharmacodynamic determinations were made after each phase of the study. The main outcome measures were serum concentrations of terfenadine and terfenadine acid metabolite, and corrected QT intervals as determined by 12-lead electrocardiogram. Significant changes occurred in time to maximum concentration (t(max)) and area under the concentration-time curve (AUC) of terfenadine and terfenadine acid metabolite after addition of grapefruit juice. All participants had detectable concentrations of unmetabolized terfenadine at the end of Phase I, which were quantified in three of the six participants. Further, all participants had increased and quantifiable levels of unmetabolized terfenadine after addition of grapefruit juice that were associated with prolongation of the QT interval relative to the baseline control period without terfenadine. Grapefruit juice did not alter the elimination half-life (t1/2) of terfenadine acid metabolite. Because of the intraindividual variability in the pharmacokinetics of terfenadine, further study is needed to confirm these results. PMID- 8728350 TI - Effects of atorvastatin, an HMG-CoA reductase inhibitor, on hepatic oxidative metabolism of antipyrine. AB - Possible effects of multiple-dose administration of atorvastatin on the pharmacokinetics of single-dose antipyrine were evaluated in this drug-drug interaction study. Twelve healthy male volunteers received three 200-mg capsules of antipyrine on days 1 and 22, and two 40-mg atorvastatin tablets in the morning on days 8 through 23. Serial blood and urine samples were collected after administration of each antipyrine dose. Plasma was analyzed for antipyrine, and urine samples were analyzed for antipyrine, 4-hydroxyantipyrine, and norantipyrine by validated high-performance liquid chromatography with ultraviolet detection. Overall, antipyrine and atorvastatin doses were well tolerated in healthy volunteers. Mean antipyrine concentrations in plasma after administration of a single, oral dose of antipyrine during coadministration of multiple doses of atorvastatin were nearly superimposible on concentrations after administration of antipyrine alone. Individual and mean parameter values for plasma pharmacokinetics of antipyrine were similar in both treatment periods. Atorvastatin did not significantly alter the fraction of clearance of antipyrine in plasma that occurred by urinary excretion of 4-hydroxyantipyrine and norantipyrine. These results indicate that the recommended highest daily dose of atorvastatin has negligible effects on antipyrine pharmacokinetics and on oxidative pathways responsible for the metabolism of antipyrine. PMID- 8728351 TI - Effect of angiotensin-converting enzyme inhibition on bronchial responsiveness. AB - The effect of angiotensin-converting enzyme (ACE) inhibition on bronchial responsiveness has not been clearly established. Because ACE degrades bradykinin and substance P, inhibition of the enzyme may lead to accumulation of these potent bronchoconstrictors in the lung, potentially leading to enhanced bronchial reactivity or bronchospasm. Previous studies of the effect of ACE inhibition on airway responsiveness have yielded conflicting results. A randomized, double blind, placebo-controlled study was therefore conducted to evaluate the effect of a 14-day course of oral lisinopril (10 mg for days 1-3, 20 mg for days 4-14) on bronchial responsiveness to inhaled methacholine in a group of healthy volunteers. No significant change in methacholine responsiveness occurred in any of the participants receiving lisinopril. The mean ( +/- SD) concentration of methacholine producing a decrease in FEV1 of 20% from baseline (PC20; mg/mL) was 23.3 +/- 5.0 before the study and 23.5 +/- 4.5 at the end of the study for the lisinopril group, and 23.0 +/- 4.6 before the study and 21.8 +/- 6.9 after the study for the placebo group. The 14-day course of ACE inhibitor therapy did not enhance nonspecific bronchial responsiveness in healthy volunteers. PMID- 8728352 TI - Hypotensive mechanisms of amifostine. AB - Amifostine, a chemo- and radioprotective agent developed as adjunctive therapy for malignancies, induces hypotension after approximately 20% of patient administrations. This study examines the molecular mechanisms underlying hypotension induced by amifostine. Amifostine and its metabolite, WR-1065, induced dose-dependent hypotension in anesthetized rats that was not blocked by N(G)-methyl L arginine (L-NAME), an NO synthase inhibitor. WR-1065 but not amifostine induced concentration-dependent relaxation of isolated rat aortic rings in an endothelium-independent fashion. Relaxation was not associated with increases in cGMP or cAMP and could not be blocked by L-NAME or indomethacin. Similarly, neither amifostine or WR-1065 activated adenylyl, particulate guanylyl, or soluble guanylyl cyclases. WR-1065 relaxed rat aortic rings precontracted with norepinepherine, suggesting alpha-adrenergic blocking activity. However, neither amifostine nor WR-1065 altered the ability of prazosin or phentolamine to bind to alpha-adrenergic receptors. Further, WR-1065 had no effect on receptor-mediated increases in intracellular calcium in BAL 17 murine B lymphocytes in vitro. Thus, hypotension after administration of amifostine is mediated by WR-1065 and appears to result from direct relaxation of vascular smooth muscle. Smooth muscle relaxation induced by WR-1065 is not related to production of nitric oxide, prostaglandins, or cyclic nucleotides; alpha adrenergic receptor antagonism; or interference with receptor-dependent increases in intracellular calcium. Administration of ephedrine, an efficacious adrenergic agonist, attenuated hypotension induced by amifostine in anesthetized rats and may be useful in alleviating hypotension associated with amifostine administration in patients. PMID- 8728353 TI - Naturally-occurring anti-thymocyte autoantibody which identifies a restricted CD4+CD8+CD3-/lo/int thymocyte subpopulation exhibits extensive polyspecificity. AB - In this study, naturally-occurring, monoclonal IgM kappa anti-thymocyte autoantibodies from the neonatal inbred Balb/c mouse-derived hybridoma NMT-1 (NMT 1 mAb), previously reported to identify a restricted CD4+CD8+CD3/lo/int thymocyte subpopulation, have been shown to exhibit extensive polyspecificity. Using immunofluorescence, immunoblotting and antibody titration and competition ELISAs, NMT-1 mAbs exhibited polyspecific binding to 12 apparently structurally unrelated self and non-self antigens. The autoreactive component of the polyspecificity profile of NMT-1 mAbs encompassed reactivity to developmentally-related 14.5 and 18.3 kDa Thy-1 glycoforms expressed on a CD4+CD8+CD3-/lo/int thymocyte subpopulation. The autoreactivity profile of NMT-1 mAbs also included recognition of the heavy and light chains of mouse IgG1 and mouse cytokeratins within thymic medullary epithelium and basal epithelial cells of stratified squamous epithelium of mouse tongue, oesophagus, stomach, skin and vagina. Examination of the polyspecificity profile of NMT-1 mAbs was also undertaken using a panel of 23 antigens including heterologous proteins, phospholipids, haptens and bacterial antigens by antibody titration and competition ELISAs. Antibody titration ELISAs demonstrated that NMT-1 mAbs bound nine antigens including bovine carbonic anhydrase, ovalbumin, cardiolipin, phosphatidylserine, the haptens, DNP and FITC and the bacterial antigens including Escherichia coli beta-galactosidase and the toxoids from Corynebacterium tetani and Clostridium diphtheria. Competition ELISAs, based on the inhibition of NMT-1 mAb binding to antigens adsorbed to ELISA plate surfaces by inhibitor antigens in solution, demonstrated that NMT-1 mAb interactions were not dependent on multivalent binding. In these assays, NMT 1 mAbs recognized unmodified (native) epitopes on the solution phase forms of the protein antigens, including E. coli beta-galactosidase and toxoids from Corynebacterium tetani and Clostridium diphtheria, providing further evidence for the hypothesis that the binding of multiple, apparently unrelated, antigens by NMT-1 mAbs occurs via unique polyspecific antigen combining sites. PMID- 8728354 TI - IL-1, ICAM-1, LFA-3, and hydrocortisone differentially regulate cytokine secretion by human fetal thymic epithelial cells. AB - Human fetal thymic epithelial cells (TEC) were cultured under serum-free conditions. The TEC were analyzed for the secretion of 14C-labelled peptides and of IL-3, IL-6 IL-7, IL-8, GM-CSF, LIF, fibronectin and thymosin alpha 1 by ELISA tests. IL-3 and IL-7 were not detected from these TEC. Lack of IL-7 and presence of thymosin alpha 1 and of the surface molecule TE-4 depicts these cells as subcapsular/medullary TEC. TEC secreted constitutively IL-6, IL-8, fibronectin and thymosin alpha 1 but not GM-CSF of LIF. Stimulants included recombinant IL-1 and monoclonal antibodies to the surface adhesion molecules ICAM-1 and LAF-3. In addition, hydrocortisone (2.7 x 10(-7) M) was used to dissect secretion patterns. Recombinant IL-1, anti ICAM-1, and anti LFA-3 alone and collectively induced modest but significant increases in the secretions of 14C-labelled peptides and of IL-6 and IL-8 which were not inhibited by HC. Recombinant IL-1 but not anti ICAM-1 and anti LFA-3 induced GM-CSF and LIF. HC inhibited the secretion of GM CSF and LIF induced by IL-1. None of the stimulants augmented the constitutive secretion of fibronectin or thymosin alpha 1 and HC inhibited thymosin alpha 1 secretion. TEC secretion of cytokines but not thymosin alpha 1 and fibronectin appear to be regulated in a more complex manner than heretofore recognized. PMID- 8728355 TI - Cyclic adenosine monophosphate (cAMP) differentially regulates IL-4 in thymocyte subsets. AB - It was recently reported that cAMP upregulated IL-4 in T cell lines generated in vitro and expressing an unrestricted lymphokine pattern. In order to study the effect of cAMP on IL-4 released by freshly isolated T cell populations, we tested various T cell subsets that have previously been reported to synthesize this lymphokine. We found that cAMP upregulated IL-4 release from in vivo activated single CD4+ peripheral T cells and CD4+CD8-HSAlowNK1.1- thymocytes stimulated with ionomycin and phorbol ester. Furthermore, as in conventional single CD4+ or CD8+ thymocytes, cAMP enhanced IL-4 production in CD4+CD8-NK1.1+ thymocytes. This latter subset has recently been shown to belong to a independent T cell lineage that is positively selected by MHC class I molecules, recognizes CD1 and expresses a restricted T cell receptor repertoire. In contrast, cAMP appeared not to upregulate IL-4 in a third independent T cell lineage. This was suggested by the observation that cAMP did not increase IL-4 in CD3+CD4-CD8- thymocytes, which are thought not to give rise to conventional single CD4+ or CD8+ T cells. It therefore appears that cAMP is coupled differently to IL-4, depending on the T cell lineage. In contrast, in all IL-4 producing stages of conventional T cells, IL-4 is consistently upregulated by cAMP. PMID- 8728356 TI - Increased numbers of CD5+ B cells in the thymus of estradiol benzoate-treated rats. AB - In the present work we combine both flow cytometry and in situ immunohistochemical techniques to study the changes affecting a minor B cell population described within the normal rat thymus, after treatment with estradiol benzoate (EB). Our results, in agreement with previous data, show that the vast majority of these intrathymic B cells are CD5+. The existence of CD5+ B cells was confirmed flow cytometrically in both cervical lymph nodes and spleen of control, adult Wistar rats. Moreover, after EB administration intrathymic B cells increased significantly especially in those rats receiving 500 micrograms of EB, constituting cell masses around the blood vessels of cortico-medullary area and in the thymic medulla. We discuss the significance of this increased number of intrathymic CD5+ B cells, which is probably due to a selective cell migration from the periphery into the thymus, from the view of the effects of estradiol on the thymic vascular permeability. PMID- 8728357 TI - Attitude of African-Americans regarding prostate cancer clinical trials. AB - The purpose of this study was to qualitatively assess attitudes associated with the willingness of African-Americans to participate in prostate cancer clinical trials. Fifty-six African-American males, 40 years of age and older, were recruited from South Central Los Angeles. Respondents were divided into lower or middle socio-economic groups based on education and occupation. Focus group discussions were conducted to assess their knowledge about prostate cancer and willingness to participate in prostate cancer clinical trials. In addition, information was obtained to identify their incentives and barriers towards participating in prostate cancer research. Middle socio-economic respondents expressed a greater willingness to participate in prostate cancer clinical trials than did men of lower socio-economic status. Many indicated that they would be more likely to participate if they were encouraged to do so by a physician or researcher who was viewed as being competent and compassionate. Barriers to participation in prostate cancer clinical trials included concerns about drug toxicity, medical experimentation and distrust of the medical establishment. Endeavors aimed at increasing minority representation in prostate cancer clinical studies should address these issues. PMID- 8728358 TI - Perceptions of cervical cancer and Pap smear screening behavior by women's sexual orientation. AB - This study examined 330 adult females' perceptions and practices regarding cervical cancer by sexual orientation. Ninety-four percent of respondents were unable to correctly identify all 5 risk factors associated with increased risk for cervical cancer (smoking, sexual intercourse with men, multiple male sexual partners, sexual intercourse before age 16, and having genital warts) regardless of sexual orientation. Furthermore, 20% were not able to identify any of the 5 risk factors for cervical cancer. Lesbians perceived themselves to be less susceptible to cervical cancer than heterosexuals or bisexuals even though 79% of lesbians had sexual intercourse with a male. Eight percent of the respondents believed that most women who develop cervical cancer die from the disease. The main barriers identified by non-regular Pap screeners (N = 127) were: no health insurance (33%), forgetting to get a pap test (32%), and not liking to get a Pap test (31%). Three-fourths (75%) have had a Pap test within the last 2 years and planned to continue having them on a regular basis. The results of this survey indicate that there is considerable room for improvement in knowledge, perceptions, and practices of all women, regardless of sexual orientation, regarding cervical cancer. PMID- 8728359 TI - Diet and exercise profiles of 30- to 60-year-old male smokers: implications for community heart health programs. AB - The aim of this study was to estimate the prevalence of diet and exercise behaviors in middle-aged male smokers and identify their determinants of behaviors, with the ultimate goal of designing more effective interventions in a community heart health program. Self-administered, postal questionnaires produced a sample of n = 671 male smokers, 30 to 60 years of age residing in Laval, Quebec's second most populous city. Variables for the study were derived from the theory of planned behavior. Correspondence analysis explained 65% of the variance, classifying smokers into five groups based on diet and exercise behavior. Group 1, smokers with diet and exercise behaviors deemed inadequate for heart health, comprised 43% of the total. Groups 2 and 3, smokers with a diet deemed inadequate or needing improvement respectively, and Group 4, smokers with inadequate exercise, comprised 42% of the total. Group 5, smokers with neither of the other high-risk behaviors, comprised 15% of the total. Four significant variables contrasted Groups 1 to 4 with group 5: age, number of cigarettes smoked daily, and perceived behavioral control (PBC) in both diet and exercise. However, coefficients for age and smoking were weak for all groups. Coefficients for PBC in diet were high and negative only for smokers with an inadequate one. For PBC in exercise, they were high and negative only for smokers who exercised inadequately. Intervention groups based on specific deficiencies in diet and exercise behaviors were then identified. Therefore, the results suggest that smoking cessation programs may be made more effective by targeting specific sub groups and by providing resources and opportunities to counteract the lack of perceived behavioral control in middle-aged male smokers vis-a-vis diet and exercise. PMID- 8728360 TI - An examination of differential follow-up rates in breast cancer screening. AB - This study was designed to test the hypothesis that follow-up rates for women with abnormal breast screening results would vary by age, ethnicity and initial screening results in California's Breast Cervical Cancer Control Program. The sample was composed of women in this screening program who had received abnormal clinical breast exam (N = 5239) or mammography results (N = 1202). Chi-squared analysis showed significant differences by age and initial screening results but in logistic regression analysis the only demographic variable to retain significance in analysis was age, although size of the clinic was also highly significant. Older women should be targeted as a group needing assistance with compliance with follow-up recommendations. PMID- 8728361 TI - A mandatory smoking ban in restaurants: concerns versus experiences. AB - The purpose of this study was (a) to assess the concerns of restaurant representatives about a city ordinance that prohibited smoking in all restaurants prior to its enactment and (b) to determine if these concerns were realized 15 months after the ordinance had been in effect. Representatives from 34 randomly selected restaurants participated in both the pre- and post-interviews. Although 26.5 percent of the respondents were concerned that the ordinance would be difficult to enforce, 94 percent found the ordinance easy or very easy to enforce. While some customers appeared to have negative reactions to an ordinance that prohibited smoking, four times that many appeared to have positive reactions to the ordinance. Although approximately 12 percent of the respondents indicated that the ordinance had "a slightly negative effect on employees", the majority (88.2%) felt that the ordinance had either no effect or a positive effect on employees. Most respondents believed that the ordinance would have no effect on their business and most reported that the ordinance had no or no know effect on business. Although many restaurant representatives had concerns about a non smoking ordinance prior to its enactment, restaurant representatives' self reported experience with the ordinance suggests that most of these concerns were not realized. PMID- 8728362 TI - Clinical practice groups. Loss of orthopedic autonomy. PMID- 8728363 TI - Biologic markers of arthritis. AB - Biologic markers of arthritis are elements detectable in synovial fluid, serum, or urine, that may reflect the underlying degenerative joint disease. They have the potential to be highly sensitive and specific to the presence of subclinical joint disease. Accordingly, they have applications in the early diagnosis of arthritis, the close monitoring of disease progression, and the prediction of the patient's response to treatment. There are four major categories of substances being investigated as potential biologic markers of arthritis: constituents of the extracellular matrix, degradative proteolytic enzymes, cytokines, and nitric oxide. This article reviews the state of the current research in this emerging field and its potential clinical applications. PMID- 8728364 TI - Rotational stability of noncemented total hip femoral components. AB - Achieving torsional fixation of the femoral component in total hip arthroplasty is an important factor in success of the implant, so design features that improve fixation are likely to improve clinical results. Four femoral stem designs that allow different levels of femoral neck resection and use different distal fixation techniques were mechanically tested in cadaveric femurs to determine resistance to torsional loads. Five specimens for each stem design were implanted according to the standard procedure, and each was axially and torsionally loaded in a servohydraulic testing machine. Rotational interface micromotion and interface slippage were measured at the bone-implant interface. Decreased micromotion and interface slippage were associated with a distal scratch fit and neck retaining design, and the combination of distal scratch fit and neck retention had the least micromotion and smallest standard deviation of the four designs. PMID- 8728365 TI - Early experience with eccentric acetabular components in revision total hip arthroplasty. AB - Osteolysis associated with acetabular component loosening can produce significant acetabular defects. The combined (segmental and cavitary) or type III defect can present a very challenging reconstructive problem. Several methods of reconstruction for type III defects have been described, including the use of oblong or eccentric sockets. These implants are attractive, theoretically, because they allow restoration of the hip center without use of structural bone graft. The results of this method however have not been widely reported. From a series of type III defects reconstructed from 1987 to 1991, six hips were reconstructed with eccentric acetabular components, provided by three different manufacturers. Four of these six implants were custom and two were off-the-shelf. At 4-years minimum follow-up, three of the six sockets had failed and had been revised. All three failures had been custom implants produced by the same manufacturer. The other three sockets had produced good clinical results. The features common to the failed implants were: (1) custom manufacture; (2) small ingrowth pads not integral with the body of the implant; and (3) lack of specific instrumentation (reamers) for insertion of the implant. Eccentric sockets may have limited, although well-defined, indications for use, such as acetabular revision needed in the presence of a well-fixed or monolithic femoral stem. Early experience suggests that custom eccentric acetabular components are not well suited for such indications. PMID- 8728366 TI - Simultaneous bilateral total knee replacement versus unilateral replacement. AB - Over a 5-year period the two authors performed 213 simultaneous (sequentially) bilateral total knee arthroplasties (426 knees). A matched series of 107 unilateral arthroplasties was performed within this time frame, and the two groups of patients were evaluated based on postoperative complications and functional improvements. The patients in both groups were primarily osteoarthritics with an average age of 70 years. Knee function according to Hospital for Special Surgery (HSS) scores showed no significant difference, with the postoperative scores for the bilaterals and unilaterals averaging 93.6 and 93.4, respectively. The complication rate in the bilateral series was not increased over the unilaterals; it was less. These results lead to the conclusion that in patients with bilateral arthritis, both knees should be operated on under a single anesthetic, providing the surgical team is able to carry out the operative procedure expeditiously. PMID- 8728367 TI - Patellar tilt in total knee arthroplasty. AB - Eighty-nine consecutive primary total knee arthroplasties performed for osteoarthrosis were evaluated to determine the effect of patellar thickness, alignment, joint line, patellar height, type of patellar prosthesis, and length of follow-up on patellar tilt. Preoperatively, 62% of the patellae were tilted neutrally and 38% tilted laterally on standard 45 degrees Merchant-view radiographs. At 3-week follow-up, 52% were neutral, 32% tilted laterally and 16% tilted medially. At the most recent follow-up (mean 36 months), 41% were neutral, 46% tilted laterally and 13% tilted medially. Five (5.6%) required reoperation for patellar complications. Change in joint line, in alignment, and in patellar height had no effect on postoperative patellar tilt. Patients who required a lateral release had postoperative patellar tilt measurements that were not statistically different from those who did not require a lateral release. Metal back domed patellae were more likely to tilt laterally than all-polyethylene domed patellae; however, the metal-back patellae had a longer mean follow-up (41 months) compared with the all-polyethylene patellae (27 months). The positive correlation between preoperative patellar thickness and postoperative patellar tilt was statistically significant. The negative correlation between change in patellar thickness and postoperative patellar tilt also was statistically significant. Finally, postoperative medically tilted patellae moved toward neutral with time and laterally tilted patellae not only remained laterally tilted but also the tilt increased with time. PMID- 8728368 TI - Lisfranc dislocation and associated metatarsophalangeal joint dislocations. A case report and literature review. AB - A case report of a 28-year-old man with associated dislocations of the tarsometatarsal (Lisfranc) and metatarsophalangeal joints is presented. The potential for disability after these injuries is very high when the diagnosis or treatment is delayed, the reduction is incomplete, or the dislocation recurs. Relevant aspects of diagnosis and treatment are discussed. PMID- 8728369 TI - Posterior dislocation of a posterior-stabilized total knee arthroplasty. A report of two cases. AB - Two patients had posterior dislocation of a posterior-stabilized total knee arthroplasty. In both cases, the patients required general anesthesia during the procedure to unlock the components and reduce the dislocations. The surgical technique and design considerations are discussed. PMID- 8728370 TI - Medial dislocation of the long head of the biceps tendon. Magnetic resonance imaging evaluation. AB - Medial dislocation of the long head of the biceps branchii tendon is a rare occurrence and is often associated with degenerative or traumatic tears of the rotator cuff, specifically tears of the subscapularis tendon. Following a dislocation, the biceps tendon will assume either an intra- or extra-articular position depending on whether or not the subscapularis tendon detaches from its humoral insertion. Magnetic resonance imaging (MRI) has been found to provide valuable information concerning the location of the biceps tendon and the integrity of the rotator cuff. In this report, three patients with suspected dislocations of the biceps tendon are evaluated using MRI. PMID- 8728371 TI - Referrals for specialized care are anathema to capitation systems. PMID- 8728372 TI - A 45-year old woman with right shoulder pain. PMID- 8728373 TI - Ganglion of the anterior cruciate ligament presented as a mass. PMID- 8728374 TI - Current issues in the management of patients with cytomegalovirus infection. PMID- 8728375 TI - Current and experimental therapeutic options for cytomegalovirus disease. PMID- 8728376 TI - Current management of cytomegalovirus infection: treatment and prophylaxis. PMID- 8728377 TI - Cytomegalovirus resistance to antiviral therapies. PMID- 8728378 TI - Second protease inhibitor approved for HIV therapy. PMID- 8728380 TI - Health systems computerizing on massive scale. PMID- 8728379 TI - Gastroenterological associations recognize three antimicrobial combinations for ulcer therapy. PMID- 8728381 TI - Oral morphine eases chronic non-cancer pain, with little risk of addiction. PMID- 8728382 TI - Behavioral pharmaceutical care scale for measuring pharmacists' activities. AB - The development and validation of a behavioral pharmaceutical care scale (BPCS) is described. The BPCS items were constructed by conducting an extensive review of the literature on pharmaceutical care and a focus group meeting. To validate the instrument, data were collected from 617 community pharmacists in Florida. Reliability coefficients for the BPCS domains were > 0.70, and the content validity index value for the whole instrument was 0.79. Evidence supporting trait validity of the BPCS was provided by using confirmatory factor analysis to confirm the instrument's dimensionality. Nomologic validity was established by confirming the hypothesis that pharmacists who in a prior survey reported they intend to provide pharmaceutical care would have a significantly higher BPCS score than those who reported they do not intend to provide pharmaceutical care. The behavioral pharmaceutical care scale, developed as a tool for measuring pharmacists' efforts to provide pharmaceutical care, was found to be reliable, sensitive, and valid. PMID- 8728383 TI - Promoting use of a preferred histamine H2-receptor antagonist in managed care organizations. AB - A program to promote generic cimetidine as the preferred histamine H2-receptor antagonist (HRA) in the managed care organizations (MCOs) served by a pharmacy benefit management company is described. A pharmacy benefit management company conducted a literature review to substantiate the therapeutic equivalence of the HRAs and to set conservative criteria for identifying candidates for conversion to oral cimetidine therapy. During the third quarter of 1994, the prescriber for each patient identified was sent a document listing the patient's current HRA therapy, the date of his or her last prescription refill, and the name and telephone number of the dispensing pharmacy. A letter summarized the literature; outlined the criteria used to identify candidate patients; gave the current indications, dosages, and average wholesale prices of the HRAs; and asked the prescriber to switch the patient to generic cimetidine. HRA use in an MCO that participated in the program was compared with HRA use in a nonparticipating MCO for the second quarter of 1994 (the baseline period), the fourth quarter of 1994, and the first quarter of 1995. The nonparticipating MCO showed no change between baseline and 1995 in the proportion of HRA prescriptions accounted for by brand name and generic cimetidine combined (14% for each period). The average acquisition cost per HRA prescription remained about $75 for each study quarter. In the participating MCO, the proportion of HRA prescriptions accounted for by brand-name plus generic cimetidine increased from 18% at baseline to 39% in the first quarter of 1995. The average acquisition cost per HRA prescription fell from $71 at baseline to $65 in the first quarter of 1995. A program to shift the use of brand-name HRAs to generic cimetidine in MCOs successfully altered prescribing patterns and reduced expenditures for these agents. PMID- 8728384 TI - Selecting infusion devices for use in ambulatory care. AB - Intravenous infusion devices commonly used in home care and ambulatory care settings are reviewed and factors to consider in selecting a device are suggested. The type of therapy to be administered, the patient or caregiver's ability to understand and carry out instructions, staff time required for patient teaching and drug and device preparation, drug stability, frequency of doses, reservoir volume, control of flow rate, type of venous access, cost and availability of devices and supplies, and reimbursement should be considered. Cost-effectiveness of a device can be evaluated only by analyzing all of the costs associated with administering a medication. Decisions must be based on an individual agency's needs, but usually one type of single-dose infuser and one brand of electronic ambulatory-care infusion pump can meet the needs of most of an agency's patients. For patients self-administering up to four doses per day, appropriate methods may include slow intravenous injection (i.v. push), infusion from minibags and tubing, and the use of elastomeric infusers, electronic or mechanical syringe pumps, or a new device based on infusion across a bioelectric membrane. Some of these types of infusers can also be used for continuous infusion. Syringe pumps are reliable and affordable and control infusion rates well, but infusion volume is limited and these devices must be recovered, cleaned, and tested between patients. Although elastomeric pumps may have higher price tags than other devices, they are simple for patients to use and dispose of. While elastomeric devices have tubing permanently attached, the disposable tubing of some other devices is detachable and can be reused if institutional policy permits. Electronic ambulatory-care infusion pumps can meet a wide range of infusion requirements. They vary in size, weight, ability to detect occlusions, features, and reliability. All use proprietary infusion sets, but the costs of sets and disposable supplies vary. Some pumps can be used for a single infusion mode, such as patient-controlled analgesia (PCA); others offer two or more modes (for example, continuous, intermittent, and PCA). "Multichannel" pumps can be used for simultaneous infusion of up to four medications; the rate of each infusion is programmed separately, and some multichannel devices offer multiple infusion modes. Some can be programmed remotely by telephone. Before an agency decides on a pump, it can investigate current users' and rental firms' experiences with the device. Also, all relevant personnel should try programming the device. Determining which devices to use requires a comparison of features that are pertinent to the particular agency or institution and a cost analysis that considers acquisition, reimbursement, patient training time, and the cost of disposable supplies. PMID- 8728385 TI - Survey of board-certified pharmacotherapy specialists in Minnesota. PMID- 8728386 TI - Additional anticoagulant strategies for managing heparin-induced thrombocytopenia. PMID- 8728387 TI - A phylogenetic analysis of cuckoo bumblebees (Psithyrus, Lepeletier) and bumblebees (Bombus, Latreille) inferred from sequences of the mitochondrial gene cytochrome oxidase I. AB - PCR amplification and direct sequencing of a 532-bp region of the mt CO-1 (cytochrome oxidase I) gene from five true bumblebee species and six cuckoo bumblebee species were performed. The sequences were then aligned to the corresponding sequence in the honey bee. Phylogenetic analyses based on parsimony and maximum likelihood indicate that the cuckoo bumblebees form a monophyletic group within the true bumblebees. The Bombus lucorum group (subgenus Bombus) falls out as the sister group to a clade comprising the cuckoo bumblebees and the remaining true bumblebees. A strong A + T bias (A + T = 75%) was recorded. Comparisons of all the sequences show a high-transversion bias (A<-->T) even among closely related species. The bias is most pronounced in third positions. PMID- 8728388 TI - Evolution of stomach lysozyme: the pig lysozyme gene. AB - The acquisition of an efficient stomach lysozyme is associated with the success of the ruminants. Advanced ruminants, such as cow, sheep, and deer, have approximately 10 lysozyme genes, some of which are expressed and function in the stomach and some which are expressed and function in nonstomach tissues (e.g., trachea or kidney). The pig possesses a single conventional lysozyme c gene that is expressed in both stomach and nonstomach tissues, and in this respect is similar to what was thought to exist in the early artiodactyl, before the acquisition of the ruminant lifestyle. To better understand the genetic events that occurred early in the origin and evolution of stomach lysozyme, we have isolated and characterized the pig lysozyme gene. The pig lysozyme gene is similar in size to that of other mammalian species, and both stomach and nonstomach expression utilize the same promoter. All the duplications of the ruminant lysozyme gene occurred after the divergence of the pig lineage from the lineage leading to the advanced ruminants. Comparison of the nucleotide sequence of the coding region of mature stomach lysozymes from advanced ruminants and pig revealed no change in the rate of synonymous substitutions. Comparison of the numbers of nonsynonymous and synonymous substitutions provides evidence for positive selection along the early ruminant lineage. These results indicate that changes in selective pressure, and not mutation rate, account for the changes in rates of stomach lysozyme evolution. PMID- 8728389 TI - Evidence for four Hox clusters in the killifish Fundulus heteroclitus (teleostei). AB - We analyzed the HOM/Hox cluster composition of the teleost Fundulus heteroclitus (Killifish) using a PCR survey. We found a total number of 30 unique homeobox sequences which could be assigned to specific cognate groups of the Hox clusters by sequence comparisons. One sequence was of the msh group. Ten homeobox fragments could be identified as orthologs with specific mammalian Hox genes of cluster A-D. The number of representatives in cognate groups 1, 4, 5, 6, 7, 8 and 9 differed from those of human and mouse. For groups 1 and 9 we found four representatives, which provides the first evidence for possible four Hox clusters in a diploid teleost, a lower vertebrate. Furthermore, it demonstrates significant differences of the Fundulus clusters compared to those of mouse and humans. The implications of our data for the interpretation of the evolution of Hox clusters are discussed. PMID- 8728390 TI - Complete sequence of the mitochondrial DNA in the sea urchin Arbacia lixula: conserved features of the echinoid mitochondrial genome. AB - The complete nucleotide sequence (15,719 nucleotides) of the mitochondrial DNA (mtDNA) from the sea urchin Arbacia lixula is presented. The comparison of gene arrangement between different echinoderm orders of the same class provides evidence that the gene organization is conserved within the same echinoderm class. The peculiarities of sea urchin mtDNA features, already described, are confirmed by the A. lixula mtDNA sequence. The comparison of the entire sequences of mtDNA among A. lixula, Paracentrotus lividus, and Strongylocentrotus purpuratus allowed us to detect peculiar features, common to the three sea urchin species, that can represent the molecular signature of the mt genome in the sea urchin group. Analysis of the nucleotide composition indicates that A. lixula mtDNA, in contrast with the mtDNA of other sea urchins, shows a bias in the use of T and tends to avoid the use of C, most evident in the neutral part of the molecule, such as the third codon positions. This observation indicates that the three sea urchin mtDNAs evolve under different mutation pressure. Analysis of the sequence evolution allowed us to confirm the phylogenetic tree. However, the absolute divergence time, calculated on the basis of paleontological estimates, largely diverged from the expected one. PMID- 8728391 TI - The analysis of protein coding genes suggests monophyly of Trypanosoma. AB - We analyze evolutionary relationships among members of the family Trypanosomatidae, with particular emphasis on whether protein coding genes support paraphyly of the genus Trypanosoma. Phylogenetic reconstruction based on three different protein coding genes (glyceraldehyde-3-phosphate dehydrogenase, trypanothione reductase, and alpha-tubulin) suggests that Trypanosoma is monophyletic. Moreover, pairwise comparisons of other protein coding genes show that the distances between Trypanosoma cruzi and T. brucei are significantly smaller than are the distances between each Trypanosoma species and Crithidia or Leishmania. These results contradict recent published phylogenies based on nuclear rRNA genes which suggested that T. cruzi is more closely related to Leishmania and Crithidia than to T. brucei. PMID- 8728392 TI - Mitochondrial gene order adjacent to the control region in crocodile, turtle, and tuatara. AB - We used the polymerase chain reaction and sequencing of mitochondrial gene junctions adjacent to the 5' end of the control region (light strand orientation) and the 3' end of ND6 to assess whether a representative crocodilian, turtle, or tuatara shares a unique mitochondrial gene order that is found in birds but not in mammals or amphibians. Turtles and crocodiles have the same gene order as mammals, except that crocodile has a tRNAPhe gene inserted between tRNAPro and the 5' end of the control region. Two different arrangements were detected at the 5' end of the control region in the tuatara, one resembling the mammalian (but with tRNAThr absent) and one resembling the avian gene order. Our data are consistent with the hypothesis that some tuatara mtDNAs within a single individual have undergone a deletion that removed the genes coding for cytochrome b and tRNAPro as well as 87 bp of the control region. PMID- 8728393 TI - Molecular phylogenetics of Poaceae: an expanded analysis of rbcL sequence data. AB - Phylogenetic analyses of rbcL sequence data from grasses are presented, extending the work of an earlier study. Based on molecular evidence that Joinvilleaceae and Restionaceae are closely allied to Poaceae, species from these families and from a distantly related dicot were variously employed as outgroups and analyzed with 23 species of Poaceae using the parsimony method. Measures of support were determined. The terminal portion of the tree was invariant in all analyses. Outgroup selection affected the basal topology of the tree with somewhat different measures of support for the deeper branches of the tree. When the outgroup was the dicot, pooid grasses made up the basal clade. When the outgroup was Joinvilleaceae or Restionaceae, species of Bambusoideae were basalmost. Increased sampling did not result in a marked increase in support of the rbcL data for the earliest branching events. PMID- 8728394 TI - Phylogenetic analysis of beta-tubulin sequences from amitochondrial protozoa. AB - It has been proposed that certain extant anaerobic protozoa are descended from organisms that diverged early in eukaryotic evolution prior to the acquisition of mitochondria. Among these are the extracellular parasites Giardia lamblia, Trichomonas vaginalis, and Entamoeba histolytica, and the obligately intracellular microsporidia. Phylogenetic analysis of rRNA sequences from these amitochondrial organisms suggests that G. lamblia, T. vaginalis, and microsporidia are near the base of the eukaryotic tree, while E. histolytica clusters with mitochondria-containing species. However, since eukaryotes likely evolved by symbiotic associations, it is important to analyze other sequences which may have independent origins. Unlike ribosomes, microtubules appear to be unique to eukaryotes. Complete gene sequences for the beta-tubulin subunit of microtubules from T. vaginalis, E. histolytica, and the microsporidian Encephalitozoon hellem have recently been determined. Phylogenetic relationships among these, G. lamblia, and 20 additional beta-tubulins were analyzed by distance matrix and parsimony methods, using alpha- and gamma-tubulin outgroups. All analyses placed the E. histolytica sequence at the base of the beta-tubulin evolutionary tree. Similar results were obtained for E. histolytica alpha-tubulin using a less representative set of sequences. In contrast, the E. hellem sequence branched considerably higher, within the lineage containing animal and fungal beta-tubulins. Possible explanations are considered for these unexpected differences between the beta-tubulin and rRNA trees. PMID- 8728395 TI - Molecular phylogenetics and biogeography of tanagers in the genus Ramphocelus (Aves). AB - Ramphocelus tanagers are distributed throughout the Neotropical lowlands. In this paper, mitochondrial DNA (mtDNA) sequence data from cytochrome b and ND2 genes are used to estimate relationships among seven of nine species of the avian genus Ramphocelus. Genetic differentiation is high between Ramphocelus passerinii passerinii and Ramphocelus passerinii costaricensis, and the two subspecies are diagnosable and distinct from one another both morphologically and genetically. Thus, elevation to species status is recommended. Three clades are supported by both gene sequences; one clade contains R. passerinii, R. costaricensis, and R. icteronotus; the second clade contains Ramphocelus carbo, Ramphocelus bresilius, and Ramphocelus nigrogularis; the third clade contains Ramphocelus sanguinolentus. The degree of saturation was assessed for both genes and saturation of third position of codons occurs by 10-12% uncorrected pairwise sequence divergence. The general area cladogram suggests the following area relationships: Pacific and Caribbean Central America are sister areas, Choco is the sister to the Central American area, and Amazonia/southeastern Brazil is the outgroup area to the Choco/Central American clade. PMID- 8728396 TI - Mitochondrial phylogeny of trematomid fishes (Nototheniidae, Perciformes) and the evolution of Antarctic fish. AB - The subfamily of fishes Trematominae is endemic to the subzero waters of antarctica and is part of the longer notothenioid radiation. Partial mitochondrial sequences from the 12S and 16S ribosomal RNA (rRNA) genes and a phylogeny for 10 trematomid species are presented. As has been previously suggested, two taxa, Trematomus scotti and T. newnesi, do not appear to be part of the main trematomid radiation. The genus Pagothenia is nested within the genus Trematomus and has evolved a unique cyropelagic existence, an association with pack ice. Using a mitochondrial rRNA molecular clock rate of 0.14% transversion changes per million years, the average age of the trematomids is estimated at 3.4 million years (MY). If the age of the trematomids is approximately 3.4 MY, this group could have speciated during the period of deglaciation in Antarctica 2.5 4.8 million years ago. This era was marked by significant changes on the Antarctic shores, such as the opening of fjords, which might have provided a stimulus for specification. PMID- 8728397 TI - Phylogeny of the Chlamydomonadales (Chlorophyceae): a comparison of ribosomal RNA gene sequences from the nucleus and the chloroplast. AB - Phylogenetic analyses of nuclear-encoded small-subunit rRNA sequences and chloroplast-encoded large-subunit rRNA sequences from flagellate green algae representing the order Chlamydomonadales were found to show considerable congruence. In general, the chloroplast data set exhibited more robust support for comparable lineages than the nuclear data set. The phylogenetic inferences derived from the independent data sets support some, but also challenge many, traditional taxonomic and phylogenetic concepts regarding the green flagellates. Results from phylogenetic analyses of both molecular data sets support six distinct lineages that include taxa from the biflagellate genus, Chlamydomonas, and a basal lineage that comprises taxa from the quadriflagellate genus, Carteria. Both data sets support the conclusion that Chlamydomonas is not monophyletic. Although the chloroplast data are ambiguous regarding the question of Carteria monophyly, the nuclear data fail to support Carteria monophyly. The chlorococcalean genus Chlorococcum was found to have affinities with the Chlamydomonadales, indicating that the traditional concepts of both Chlorococcales and Chlamydomonadales may need revision. The genus Dunaliella is allied within the Chlamydomonadales, supporting the contention that it has lost a typical glycoprotein cell wall. PMID- 8728398 TI - The simultaneous diversification of South American echimyid rodents (Hystricognathi) based on complete cytochrome b sequences. AB - Variation in the complete nucleotide sequence of the mitochondrial cytochrome b gene was examined for 32 individuals representing 12 supraspecific taxa of South American rodents of the family Echimyidae (Hystricognathi). Representative genera of four other New World hystricognath families, the Old World porcupine Hystrix, and the myomorph murid rodents Rattus and Mus were used as outgroups in phylogenetic reconstructions. Monopoly of the family Echimyidae is strongly supported, a result fully consistent with existing morphological and paleontological data relative to the taxa examined. However, relationships among most supraspecific taxa within the family are poorly resolved. Poor resolution appears not to result from lack of data, but to a rapid, nearly simultaneous divergence of most Recent taxa. Generic groupings that are moderately to strongly supported include the tree rats of the Brazilian Atlantic Forest (Nelomys) and Amazonia (Echimys, Makalata) and the Amazonian arboreal spiny rats Mesomys and Lonchothrix. However, the two subgenera of the terrestrial spiny rats, Proechimys, do not form a monophyletic unit, and elevation of the Atlantic Forest Trinomys to generic status is supported. The genus Hoplomys is closely related to Proechimys (sensu stricto), a finding supported by other molecular data. PMID- 8728399 TI - Early evolution of the Metazoa and phylogenetic status of diploblasts as inferred from amino acid sequence of elongation factor-1 alpha. AB - To understand the early evolution of the Metazoa, it is necessary to determine the correct phylogenetic status of diploblastic animals (poriferan, cnidaria, and ctenophora). Despite clasdistic studies of morphological characters and recent molecular phylogenetic studies, it remains uncertain whether diploblasts are monophyletic or paraphyletic, and how these three phyla of diploblasts are phylogenetically related. To obtain insight into these phylogenetic problems, we sequenced almost the entire nucleic acid sequence of elongation factor-1 alpha from a sponge, two cnidarians, a ctenophora, and a turbellarian. We then investigated the phylogenetic status of the diploblasts. We compared the amino acid sequences, nucleotide sequences at the first and second codon positions, and those at the second positions. Phylogenetic trees were inferred by neighbor joining, maximum likelihood, and maximum parsimony, and they supported the monophyly of the Metazoa. However, the phylogenetic relationships of the diploblast groups were not significantly resolved, although the trees preferred the monopoly of the diploblasts. PMID- 8728400 TI - Molecular phylogeny of Azteca ants (Hymenoptera:Formicidae) and the colonization of Cecropia trees. AB - Despite the prominence of the Azteca-Cecropia interaction as the focus of extensive ecological investigation, a reliable phylogeny of the Azteca ants has been lacking, primarily because many of the morphological and behavioral characters are phylogenetically uninformative or conflicting. A phylogenetic analysis of a select set of Azteca ants, including six Cecropia inhabitants and two non-Cecropia inhabitants, plus an outgroup taxon, is presented on the basis of mitochondrial DNA sequences. The evolutionary relationships deduced from the molecular data are analyzed with reference to ecological and morphological studies, specifically addressing the phylogenetic relationship of structurally an behaviorally ambiguous taxa, species complex groupings, and the colonization of Cecropia trees. According to the molecular phylogeny, the Cecropia-inhabiting Azteca do not form a monophyletic clade, indicating multiple independent colonization or abandonment of Cecropia trees by the Azteca. PMID- 8728401 TI - The distribution and phylogenetic significance of a 50-kb chloroplast DNA inversion in the flowering plant family Leguminosae. AB - Species in 9 of the approximately 650 genera of the flowering plant family Leguminosae are known to possess a large (50-kb) inversion in their chloroplast genomes, relative to the gene order found most commonly among land plants. Putatively basal elements of the family have not been surveyed for the inversion, which is unknown outside the legumes. Using a combination of polymerase chain reaction and restriction-mapping approaches employing primers or hybridization probes flanking inversion endpoints, 132 legume genera were screened for the presence of the inversion. The inversion was found to be absent in all taxa from two of the three subfamilies (Mimosoideae and Caesalpinioideae), whereas the inversion was found to be present in most taxa of the third subfamily (Papilionoideae). Two papilionoid tribes, Swartzieae and Sophoreae, were heterogeneous for the inversion, which is consistent with a number of lines of evidence suggesting the polyphyly of these tribes. The 50-kb inversion appears to be a unique event in the evolution of Leguminosae, providing a synapomorphy for a clade that includes most of the Papilionoideae. PMID- 8728402 TI - Re: The questionable implications of the dopamine D4 receptor (DRD4) gene tree for primate phylogeny. PMID- 8728403 TI - High resolution evoked potentials of cognition. AB - The precision of four methods of quantifying neuroelectric signals has been improved by increasing EEG spatial sampling, using up to 124 electrodes, and by accurate anatomical registration of the EEG with Magnetic Resonance Images (MRIs). One such method, equivalent dipole modeling, is a well-known form of source localization which is useful when the generator of the scalp recorded signal approximates a simple dipolar source, as is usually the case with early and mid-latency Evoked Potentials (EPs). Two methods of enhancing spatial detail which benefit from increased spatial sampling include the Laplacian Derivation and the Finite Element Deblurring method. The latter is a new technique which estimates the EP distribution at the superficial cortical surface. The fourth method, Evoked Potential Covariance, characterizes the spatiotemporal relationships among EP segments at different recording sites. This is useful when studying "functional neural networks" underlying higher cognitive functions. These methods are reviewed and examples of results of their application in recent experiments are presented. PMID- 8728404 TI - Quantitative EEG analysis in clinical settings. AB - Quantitative EEG techniques are becoming more available. Eventually, all EEG will be digital. Various digital utility programs can help even now with storage and viewing the polygraph EEG itself. Techniques of frequency analysis, topographic mapping and discriminant functions are also available but have limited clinical use. Applications as a monitoring tool and careful analysis of epileptic spikes have been shown some promise but need further study to identify their proper clinical roles. PMID- 8728405 TI - Quantitative analysis of epileptic discharges. AB - The methods for analysing the EEG epileptiform activity are discussed and compared. Particular emphasis is placed on studying interictal spike activity recorded from scalp. The results of these analyses should be interpreted in terms of underlying physiological events. An example is the hypothetical model of benign rolandic epilepsy of childhood. PMID- 8728406 TI - Sources of spontaneous slow waves associated with brain lesions, localized by using the MEG. AB - Electric or magnetic slow wave brain activity can be associated with brain lesions. For an accurate source localization we transformed the magnetoencephalographic (MEG) coordinate system to the magnetic resonance imaging (MRI) system by using a surface fit of the digitally measured head surface and the reconstructed surface of the MRI scan. Furthermore we solved the problem to separate sources of focal activity from other multiple sources by introducing a spatial average, the Dipole Density Plot (DDP). The DDP shows in a quantified manner concentrations of dipoles across time. The DDP uses the single dipole model adequately, because only those signal sections will be analyzed, where one component contributes to the signal predominantly. In all cases, where multiple sources concurrently active are to be localized, a current distribution analysis will be used, the Current Localization by Spatial Filtering (CLSF). All source localization procedures were tested using structural brain lesions, which were verified by imaging techniques (MRI or CT), showing the results in close topographical relation to the lesions. The results so far let us assume, that the DDP and the CLSF are valuable tools to localize sources of focal spontaneous slow wave electrical brain activity. PMID- 8728407 TI - Variation of frontal P20 potential due to rotation of the N20-P20 dipole moment of SEPs. AB - We have applied the artificial neural network method to estimate the N20-P20 dipole from scalp SEP potentials, and have investigated the rotation of the dipole moment caused by the compression effect of a brain tumor (para-Rolandic tumor) adjacent to the central sulcus. The variation of the frontal P20 potential was demonstrated by the rotation of the N20-P20 dipole moment in 8 cases of para Rolandic tumor. By estimation of the rotation of the dipole moment, it may be possible to obtain preoperative information regarding the relation between the central sulcus and the tumor. PMID- 8728409 TI - Cortical and subcortical SEPs following posterior tibial nerve stimulation. AB - Intra-operative cortical and subcortical SEPs from the cerebral convexity and from the inter-hemispheric fissure were recorded following posterior tibial nerve (PTN) stimulation. Cortical and subcortical SEPs from the cerebral convexity after contra-lateral PTN stimulation consisted of N38 and P46, and their polarity reversed when the ipsi-lateral site was stimulated. On the other hand, cortical SEPs from the inter-hemispheric fissure always showed P38 and N46, whether the right or the left PTN was stimulated. Cortical and subcortical SEPs from the inter-hemispheric fissure showed clear cut polarity reversals. These findings provide good evidence for the existence of a tangential dipole oriented perpendicular to the inter-hemispheric fissure in the foot sensory area of the primary sensory cortex. SEPs recorded from the superficial part of the inter hemispheric fissure showed smaller amplitudes and longer latencies than those of SEPs from the deeper regions. These findings suggest the existence of another dipole responsible for the generation of SEPs after PTN stimulation. PMID- 8728408 TI - The neural origin generating early cortical components of SEP: topographical analysis using temporal-second-order-differentiation of cortical SEPs. AB - In order to identify dipole generators of the N20/P20 and P25, we employed second order-differentiation in the temporal dimension (temporal-second-order differentiation; TSOD) with delta t = 2 msec. The rate of variation in the voltage of cortical SEPs calculated by TSOD identified responses of each dipole, reflecting the density of neuronal firing. On topographic analysis, the distributions of N20/P20 and P25 conformed to the shape of gyrus better in the TSOD maps than in the isovoltage maps. The TSOD maps indicated that N20 and P25 were post-central components and that P20 was a pre-central one. Therefore, we concluded that the two dipoles generating N20/P20 and P25 were located in the posterior wall of the central sulcus (area 3b) and the crown (areas 1 and 2) of the post-central gyrus, respectively. PMID- 8728410 TI - Perception of visual apparent movement and topographic analysis of related evoked potentials. AB - Most of the studies in long-range apparent movement have been concerned with the psychophysical properties in central vision. However, apparent movement in peripheral vision is not clearly understood. This study focuses on the properties of the perception of apparent movement between central and peripheral vision. The results show that judgements of apparent movement in central vision indicate high performance at less than 4 degrees of spatial separation, and that judgements in peripheral vision indicate high performance at greater than 4 degrees. Evoked potentials associated with long-range apparent movement were also measured, and topographic maps were constructed to evaluate the active areas corresponding to visual apparent movement. The topographic maps show that the central regions are activated above 300 msec after the onset of the second apparent-movement stimulation. The results suggest that visual apparent movement occurs in the visual association area, or at higher levels in the central nervous system. PMID- 8728411 TI - Human VEPs to isoluminant chromatic and achromatic sinusoidal gratings: separation of parvocellular components. AB - To extract the responses specific to the parvocellular system (color vision and form perception) in humans, visual evoked potentials (VEPs) in response to the onset of isoluminant chromatic (red-green) and achromatic (black-white) sinusoidal gratings were recorded in 15 young adults. Chromatic stimulation evoked a characteristic negative wave (N1) with peak latencies around 120 msec. The amplitude resembled an inverse U-shaped function against the spatial frequency with a peak at 2 c/deg. In contrast, VEPs to achromatic gratings showed different spatial frequency characteristics with a peak at 5.3 c/deg. With variations in the luminous intensity ratio between the red and green gratings, N1 was seen to respond maximally to isoluminant stimulation. These results indicate that the combined use of isoluminant chromatic and achromatic patterns could stimulate the parvocellular system selectively in humans. PMID- 8728412 TI - Visual evoked potentials to a faint light: signal propagation analyzed with peak latency and topographic mapping. AB - Accurate localization for the process of recognition of a light stimulus is yet to be determined. We studied 19-channel VEP from nine healthy volunteers, evoked by physiologically faint light less than 200 Cd/m2, using a light emitting diode (5 mm phi, 0.3 degrees, energized for 5 msec). VEP was bandpass filtered from 0.16 to 120 Hz, and analyzed from 20 msec pre-stimulus to 184.2 msec post stimulus. The grand average VEP suggested an initial positive peak at 115.8 msec at the frontal poles and at 136 msec over the occipital areas. An initial negative peak was noted at 156 msec at the frontal poles and at 179.2 msec over the occipital areas. This might suggest that the potentials evoked had propagated from the frontal poles to the occipital areas, taking about 20 msec. However, the topographic maps contained little evidence for such a propagation, but rather indicated the waxing and waning of positive or negative extremes. PMID- 8728413 TI - The origin of pattern reversal short latency visual evoked potential as determined by dynamic topography and the dipole tracing method. AB - The generator sites of the parietal P59 and occipital N26 components elicited by hemi-field pattern reversal stimuli were investigated. The topographic distribution of the occipital N26 component showed a paradoxical lateralization, whereas that of the parietal P59 component exhibited an anatomical lateralization. The equivalent dipoles of both occipital N26 and parietal P59 components were situated on the deep mesial surface of the functioning occipital lobe. The differences in these locations were not statistically significant, but the vector moment of the parietal P59 component projected toward the functioning parieto-occipital region and one of the occipital N26 components projected away from the functioning occipital region. The generator sites of the short latency component were considered to differ from those of the middle latency visual evoked potential. Therefore both the occipital pole and the deep cerebral structure, i.e., the lateral geniculate nucleus, may play a role in the generation of equivalent dipoles. PMID- 8728414 TI - Investigation of bilateral synchronous spike-wave discharge by EEG topography. AB - To investigate the generator mechanism of the generalized discharge in childhood epilepsy, we studied its voltage field distribution using linked ear reference (L + R), common averaged reference (CA), and source derivation methods (SD). Two cases of epilepsy with clinical lapse of consciousness, and diffuse EEG spike wave burst were divided into primary bilateral synchrony (PBS) and secondary bilateral synchrony (SBS), using coherence-phase analysis. Spike topograph images were shown by three different references. The difference in spike voltage between CA and SD was also investigated (CA-SD). By all three different references, PBS spike topography showed a broad symmetrical voltage gradient. In SBS, a left frontal focal negativity was shown with a steep voltage gradient in the topographic images of spikes by L + R, CA and SD. This asymmetry disappeared gradually at the end of the spike-wave burst. Both patients showed almost identical broad symmetrical voltage gradients on CA-SD topogram. EEG topography using various references suggested that the potential fields produced by deep generators were identical for PBS and SBS, although those generated by surface generators were different. These findings support the hypothesis that SBS may be projected from the subcortical system, and activated by cortical firing. PMID- 8728415 TI - Developmental characteristics of topographic EEG in school-age children using an autoregressive model. AB - Developmental characteristics of the resting EEG were investigated in 47 school age children using statistical analysis. Total and component EEG power were statistically analyzed between the subject groups from 7 to 14 year-old using our autoregressive pattern discrimination system. In early school-age children, significant topographic differences were seen in theta waves, while in late school-age children, the differences were found in alpha waves in the frontal and occipital regions, and in beta waves in the frontal region. PMID- 8728416 TI - Smoking induced changes in EEG two-dimensional maps and blood gas analysis. AB - The effects of smoking on mental function were investigated by the two dimensional EEG mapping method, and the results were analyzed in conjunction with blood gas data that had been obtained simultaneously. Healthy medical students from the National Defense Medical College were used as the subjects for this experiment. They live in the same dormitory and a similar lifestyle to regular military personnel. Tests were carried out on two successive days, and in between the two testing sessions, each subject was requested to refrain from smoking for 24 hours. During the testing sessions, EEGs were recorded with eyes closed, before and after smoking. Two-dimensional maps were generated by unbiased polynomial interpolation for alpha activity. Various blood chemical measurements were made, among which, partial pressure of carbon dioxide (pCO2) and percentage of carbon monoxide hemoglobin (%COHb) were studied in relation to EEG changes. The smoking induced wide-spread amplitude decrease in the alpha maps. After 24 hours of withdrawal, alpha amplitude increased, although it decreased again when smoking resumed. Either the correlation between EEG amplitude and %COHb, or the correlation between EEG amplitude and pCO2, was examined in three regions (anterior: A, middle: M and posterior: P) of the two-dimensional maps in both slow and quick smokers. There were no correlations observed between changes in the EEG amplitude and changes in %COHb induced by smoking, in any of the three regions in either fast or slow smokers. However, there was a clear correlation between changes in the EEG amplitude and changes in pCO2 in quick smokers, the correlation coefficient being approximately -0.7. It is possible that this relationship was due to hyperventilation, since the same tendency was not observed in slow smokers. PMID- 8728417 TI - Correlation of EEG activities between slow-wave sleep and wakefulness in patients with supra-tentorial stroke. AB - Using topographic EEG mapping, we studied the relationships between delta activity during slow-wave sleep (SWS) and the background EEG activity during wakefulness, in 11 normal subjects and 35 stroke patients with unilateral supra tentorial lesions. Delta-1 power during SWS showed a significant positive correlation with alpha-1 power during wakefulness, in both hemispheres. Delta-1 and delta-2 power during SWS correlated positively not only with alpha-2 power, but also with delta-1 and delta-2 power during wakefulness in the affected hemisphere. these figures indicate that the amount of delta activity during SWS can be associated with that of alpha activity during wakefulness. A close negative correlation was observed between delta power during SWS and the age of the subjects in the patient group. The Barthel index showed no significant correlation with delta-1 or delta-2 power in either hemisphere in patient group. Our results suggest that delta activity during SWS may be associated with dysfunction of the cerebral cortex in stroke patients as well as in normal aged subjects. PMID- 8728418 TI - New display methods of combined topographic EEG and cerebral blood flow images in the evaluation of cerebral ischemia. AB - To evaluate complicated changes in the EEG and cerebral blood flow (CBF) in cerebral infarction, EEG was recorded under various blood pressure levels, during CBF measurement with positron emission tomography, in patients with chronic cerebral infarction. Newly developed display methods which combine topographic EEG and its significance probability mapping with CBF, clearly provided objective measures of changes in EEG parameters and CBF, during induced hypo- and hypertension. PMID- 8728419 TI - Electric field distribution of event-related potentials in stroke patients. AB - Electrical field distributions of event-related potentials (ERP's) were recorded during an auditory "oddball paradigm" and were analyzed in terms of time and space. Fourteen normal subjects and 14 chronic patients were cerebral thrombosis were studied. For the components N1 and P3 of the ERP's to target stimuli, reference-independent measures (latency, global field power, location of maximal or minimal potential, and location of centroids) were determined. Stroke patients displayed P3 abnormalities in latency, amplitude, and electrical field on the scalp. In addition, N1 electrical fields were also abnormal. These ERP abnormalities correlated significantly with the extent of mental function impairment in the stroke patients, and they improved after administration of a cerebral metabolic enhancer (Nefiracetam: DM-9384). The ERP's seemed to be sensitive in indicating the effects of the drug. These data suggest that time course analysis of the spatial distribution of the ERP electrical field might be useful for evaluation of the extent of mental function impairment and the efficacy of drugs. PMID- 8728420 TI - An event-related potential study on the impairment of automatic processing of auditory input in schizophrenia. AB - Mismatch negativity (MMN) and P300 to auditory stimuli was carried out using global field power (GFP) in 25 schizophrenics and 25 normal controls. The peak latencies of MMN and P300 were significantly longer in schizophrenics than in normal controls, and the differences in MMN were marked in the frontal regions. The peak amplitudes of MMN and P300 were significantly smaller in schizophrenics. The scalp distribution of MMN to 1050 Hz tone differed between the two groups, although that to 1100 Hz was similar to each other. The results indicate an impairment in the automatic discrimination process of auditory input in schizophrenics. PMID- 8728421 TI - Human middle latency auditory evoked magnetic fields. AB - The magnetic equivalents of SN10, Po, Na, Pa, Nb and Pb (SN10m, Pom, Nam, Pam, Nbm, and Pbm) in short and middle latency auditory evoked potentials were measured with a 7-channel DC superconducting quantum interference device (SQUID). The sources of Pom, Nam, Pam, Nbm and Pbm responses were estimated to be located in the auditory cortex, while the source of SN10m was considered to be in a deeper part of the brain. In addition, the source of Pam was estimated to be in the vicinity of the moving N100m source. The source of Pbm was considered to be in a separate area, anterior to the source of Pam and N100m, which suggested that source of Pam was located in the primary auditory cortex, while the source of Pbm was located in the secondary auditory cortex. The source of N100m was considered to spread from the primary auditory cortex to the secondary auditory cortex. PMID- 8728422 TI - Source estimation of spontaneous MEG activity and auditory evoked responses in normal subjects during sleep. AB - This study focuses on source estimation of spontaneous MEG activity and auditory evoked responses during sleep. Sources of K-complexes and auditory evoked responses were investigated by magnetoencephalograph (MEG) and electroencephalograph (EEG) measurements, simultaneously. Sources of K-complexes during stage 2 sleep were investigated. The MEG results suggested that the sources of K-complexes can be modeled by two current dipoles. Dipoles for the K complexes were estimated to be located 5 mm away from the sources of the N100 components of auditory evoked responses during wakefulness. Sources of auditory evoked responses during each sleep stage were also investigated to clarify the origins of the K-complex, the vertex sharp transient, and delta waves. Estimated dipoles for the N100 component for each sleep stage were estimated to be at slightly different locations in the auditory area. Based upon results of the MEG measurements and the EEG topographies, sources of the N330 component can be modeled by multiple current dipoles, which are seen to be distributed diffusely throughout the cerebral cortex. PMID- 8728423 TI - Source models of sleep spindles using MEG and EEG measurements. AB - MEG measurements can detect brain sources that are difficult to detect with EEG measurements. The purpose of this study was to investigate models of sleep spindles using both MEG and EEG activities that had been recorded simultaneously. The components of magnetic fields perpendicular to the surface of the head were measured using a DC-SQUID with a first-derivative gradiometer. We propose three models for sleep spindles. In the first model, the source slides into the superficial region of the head so as to be perpendicular to it's surface, and with this model, the power spectrum of the MEG is decreased. In the second model, the source slides into the deeper structures, so that it is perpendicular to the surface. Here, the power spectra of both the MEG and EEG are decreased. The third model has source perpendicular to the surface, leaning and sliding into the deeper structures. Here, the power spectrum of the EEG is decreased but that of the MEG is not. PMID- 8728424 TI - Discrepancy between brain magnetic fields elicited by pattern and luminance stimulations in the fovea: adequate stimulus positions and a measure of discrepancy. AB - A conventional equivalent current dipole estimation provides one of the quantitative measures to evaluate the discrepancy between two single-dipole-like magnetic field patterns, though there is one problem; all stimulus positions in the visual field do not necessarily contribute to the generation of a single dipole-like magnetic field. Another important problem occurs when the field pattern is complex and cannot be approximated by a dipole. This makes it difficult to evaluate the discrepancy between two magnetic field patterns by the dipole parameters. In this paper, we determined the stimulus positions adequate for generating single-dipole-like magnetic field patterns by evaluating the magnetic field's goodness-of-fit to the field generated by a single dipole. We propose to use a similarity (SIM) as a quantitative measure of the discrepancy between two complex magnetic field patterns. The SIM is defined as an angle between two magnetic field vectors. We evaluated the discrepancy between the 100 ms post-stimulus responses to pattern-reversal (Rv) stimulus, pattern-onset (Pat) stimulus, and luminance-onset (Lumi) stimulus. The following results were obtained: (1) Stimulation of some of the octants in the fovea, far from the vertical meridian, elicited a single-dipole-like magnetic field pattern at a latency of 100 ms, though stimulation of the central part of the fovea, and stimulation of the octants along the vertical meridian, did not elicit a single dipole-like magnetic field pattern; (2) The discrepancy between responses was quantitatively evaluated by the SIM even if the field patterns were complex; (3) The SIM analysis showed that the discrepancy between the responses to the Rv and the Lumi stimuli, as well as that between the responses to the Pat and the Lumi stimuli, were greater than that between the responses to the Rv and the Pat stimuli. PMID- 8728425 TI - Dipole source localization of MEG by BP neural networks. AB - The purpose of this study was to examine the usefulness of BP neural network for source localization of MEG. Since the performance of this method does not depend on the complexity of brain parameters and source models, a homogeneous brain model and a single current dipole source are assumed for convenience. Localization accuracy was examined in relation to the configuration and scale of the network. As a result, average error for position and moment estimations was within 2% while the maximum error was about 5%. It was therefore concluded that the neural network method, was useful for MEG source localization, though some improvements are still necessary. PMID- 8728426 TI - Suppression of background brain activity influence in localizing epileptic spike sources from biomagnetic measurements. AB - The origin of inter-ictal epileptic activity can be localized from the magnetoencephalogram (MEG) using the Equivalent Current Dipole (ECD) as a source model. One problem with such localizations is that in many patients, the localization of an epileptic spike source becomes inaccurate because the epileptic spikes are superimposed by pathologic brain rhythmic activities. This paper proposes to use the spatial coherence of the measured magnetic field caused by the measured magnetic field to suppress its influence in the ECD localization. In the method proposed here, the covariance matrix, which expresses the spatial coherence of the rhythmic magnetic field, is first calculated using a data portion where no spikes exist and rhythmic slow waves are evident. Then, ECD localization is performed by minimizing the least-squares cost function modified by the covariance matrix. Experiments using a computer generated ECD field and rhythmic slow waves measured from a patient suffering from complex partial epilepsy prove the basic effectiveness of the method proposed. Localizations of actual spike sources in the same clinical data are performed, and results indicating the effectiveness of the proposed method are obtained. PMID- 8728427 TI - MEG and EEG topography of frontal midline theta rhythm and source localization. AB - In this paper, we report on our study of frontal midline theta (Fm theta) activity in human subjects, recorded during mental processes such as arithmetic calculation. The Fm theta is a 6-7 Hz rhythmic wave with a duration of few seconds. The Fm theta activity is observed in the central region at the front of the head. EEGs and MEGs of Fm theta were measured simultaneously during mental calculation, and we analyzed these waveforms based on both topographic EEG maps and magnetic fields measurements. A single dipole simulated the EEG topography adequately, but there are many other dipole models which can generate a similar EEG pattern. It is difficult to estimate the source location of the Fm theta from the EEG topography alone because the EEG technique has a certain ambiguity associated with source estimation. Therefore, we considered the spatial relationships between the sources and the patterns of EEG and MEG that were simulated. Although it is not possible to obtain a unique solution for the source location of Fm theta from the EEG data alone, the simultaneous recording of MEGs from a large scalp area may result in an unambiguous solution. We therefore conclude that the simultaneous recording of both MEG and EEG data is more useful for accurate localization, than the EEG alone. PMID- 8728428 TI - Scalp topography of SEP late components in patients with supra-tentorial lesions. AB - Somatosensory evoked potentials (SEP) to simultaneous bilateral median nerve stimulation were recorded in 7 patients with unilateral brain lesions, 8 patients with degenerative dementia, and 5 normal volunteers. Right-left amplitude difference was compared from serial topographic images and the amplitude was compared at homologous electrodes. In patients with unilateral lesions, the amplitude from the frontopolar, frontal, anterior-temporal, and/or occipital electrodes was smaller on the affected side at 240 and 360 msec, regardless of whether the subjects showed sensory deficit or not. No significant laterality was seen in the primary sensorimotor areas. Distribution obtained from the patients with degenerative dementia was quite symmetrical. A significant right-left amplitude difference was seen at the anterior temporal site at 180 and 240 msec in normal controls, although an almost symmetrical distribution was obtained on the topographic images. The late components of SEP are not associated with the primary somatosensory function, but possibly with other cortical pathways. PMID- 8728429 TI - Influence of head model in biomagnetic source localization. AB - We evaluated the influence of the head model on biomagnetic source localization by utilizing a computer simulation. We localized the source of a magnetic field that was calculated using a realistic head model, and then evaluated the localization errors. It was seen that the sphere model adequately localized the dipole in cases near the sensor, but not in cases where the dipole was deeply situated. PMID- 8728430 TI - When "wire codes" predict cancer better than spot measurements of magnetic fields. PMID- 8728431 TI - Cellular telephones and health. PMID- 8728432 TI - Estimating exposure in studies of residential magnetic fields and cancer: importance of short-term variability, time interval between diagnosis and measurement, and distance to power line. AB - Validity of exposure assessment methods has been a major concern in epidemiologic studies of magnetic field exposure and cancer. We conducted a study to evaluate the relative importance of distance to power lines and calculated historical magnetic fields when estimating past magnetic field exposure. Another goal was to compare results based on various estimates of magnetic field exposure, to assess the importance of short-term variability in magnetic fields, time between diagnosis and measurement, and sources of magnetic field exposure. We used data from a Swedish case-control study of residential exposure to magnetic fields and cancer. Childhood leukemia risk was associated with calculated historical annual average magnetic fields regardless of distance, and the association with distance disappeared when both variables were entered into the same logistic regression model. Relative risks for measurements at the time of the study (contemporary annual average fields, spot calculations, and spot measurements) were all close to or below unity. The results support the hypothesis that the difference between results using historical calculations and spot measurements is explained by the time interval between diagnosis and contemporary magnetic field estimates. PMID- 8728433 TI - Air pollution and daily mortality in Amsterdam. AB - Few data are available on the association between the present low levels of air pollution in Western Europe and mortality. Daily mortality counts and the concentrations of black smoke, inhalable particles (PM10), sulfur dioxide (SO2), carbon monoxide (CO), and ozone (O3) were available for Amsterdam from 1986 to 1992. We used Poisson regression analysis to control for seasonal and other long term temporal patterns. Black smoke and PM10 were positively associated with increased risk of mortality. The relative risk for a 100-micrograms per m3 increase in black smoke on the same day was 1.19 [95% confidence interval (CI) = 1.02-1.38], and that for a 100-micrograms per m3 increase in PM10 was 1.06 (95% CI = 0.99-1.14). The relative risk for individuals over 64 years of age was higher. We found no consistent association between the levels of SO2 or CO and daily mortality, but ozone lagged 2 days was positively associated with daily mortality. The effect of particulates on acute mortality was independent of these pollutants. The results of the present study are consistent with the relation reported between particulate air pollution and daily mortality in other communities in Europe and the United States. PMID- 8728434 TI - Confounding and exposure trends in case-crossover and case-time-control designs. AB - As with ordinary studies, both case-crossover and case-time-control studies can suffer from confounding, including confounding by indication. In a case-crossover analysis, confounding by fixed (constant) characteristics is eliminated by pairing of cases to themselves, at the possible cost of introducing bias due to time trends in exposure. A case-time-control analysis can correct case-crossover results only for bias due to such time trends. If an uncontrolled confounder (such as disease severity) is present, the use of time controls can introduce new confounding, and the case-time-control results may end up either more or less confounded than the ordinary and case-crossover results. The relative confounding in the different approaches depends on details of the relations among the unmeasured confounder, the study exposure, the study disease, and any trend in these variables or their effects. Like an ordinary study, a case-time-control study must assume absence of unmeasured confounders, whether fixed or time varying. Like a case-crossover study, it must also assume absence of carryover effects and can be more prone to misclassification bias than an ordinary study. PMID- 8728435 TI - Change in paternity: a risk factor for preeclampsia in multiparas. AB - Preeclampsia, a hypertensive disorder of pregnancy, is hypothesized to be a maternal immunologic response to foreign fetal antigen derived from the father's sperm. This response may be reduced by prolonged exposure to father's antigen, such as through a prior pregnancy, which may explain why multiparas are typically at lower risk for preeclampsia than nulliparas. Since multiparas with new partners are presented with a new set of paternal antigen, we hypothesize that they would have the same elevated risk for preeclampsia and gestational hypertension as nulliparas, compared with multiparas with no change in partner. We studied 5,068 nulliparas and 5,800 multiparas, 573 of whom had new partners, selected from the Child Health and Development Studies cohort (Oakland, CA, 1959 1967). Preeclampsia was diagnosed in 3.2% of nulliparas, 3.0% of multiparas with changed paternity, and 1.9% of multiparas with no change. Compared with multiparas with no change, the adjusted odds ratio for preeclampsia among nulliparas was 2.5 [95% confidence interval (CI) = 1.8-3.5]; among multiparas with new partners, the adjusted odds ratio for preeclampsia was 1.4 (95% CI = 0.8 2.4). There was a similar pattern of results for gestational hypertension. The adjusted attributable risk of preeclampsia in multiparas associated with a change in paternity was 29%. Although these findings in part support the immunologic theory of preeclampsia, they also suggest that a subsequent pregnancy with any partner is associated with a reduced risk for preeclampsia. PMID- 8728436 TI - Job stress and preeclampsia. AB - We conducted a case-control study to determine whether job strain (or stress) during pregnancy resulted in an increased risk of preeclampsia. We compared 110 nulliparous Caucasian and African-American women who had preeclampsia with 115 healthy nulliparous controls. All subjects gave birth between 1984 and 1987 in Chapel Hill, NC. Occupation was ascertained during a telephone interview. We assigned each job title a strain (or stress) score in accordance with the occupational database developed by Karasek. Overall, 54 (49%) of the control women worked during pregnancy, 14 in high-stress jobs. Logistic regression analysis indicated a 3.1-fold [95% confidence interval (CI) = 1.2-7.8] increased risk of preeclampsia for women employed in high-stress jobs (high psychological demand, low decision latitude) and an odds ratio of 2.0 (95% CI = 1.0-4.3) for low-stress jobs compared with nonworking women, while simultaneously adjusting for age, race, family history of preeclampsia, history of hypertension in the subject's mother, gravidity, smoking during pregnancy, timing of the first prenatal visit, and type of birth control used by the couple before the pregnancy. Furthermore, working women had 2.3 times the risk of developing preeclampsia (95% CI = 1.2-4.6) compared with nonworking women. Work-related psychosocial strain increased the risk of preeclampsia in our study. PMID- 8728437 TI - Maternal caffeine consumption and spontaneous abortion: a prospective cohort study. AB - We investigated the relation between caffeine beverage consumption and spontaneous abortion in 2,967 pregnant women planning to deliver at Yale-New Haven Hospital in 1988-1992. We evaluated coffee, tea, and soda drinking in the first month of pregnancy in interviews before the end of the sixteenth week of gestation. We obtained information on 98.2% of the pregnancies (including 2,714 singleton livebirths and 135 spontaneous abortions). As compared with abstention from caffeine beverages (coffee, tea, and soda), the adjusted odds ratios for spontaneous abortion associated with consumption of 1-150, 151-300, and > 300 mg caffeine daily were 0.81 [95% confidence interval (CI)) = 0.54-1.20], 0.89 (95% CI = 0.48-1.64), and 1.75 (95% CI = 0.88-3.47), respectively. Drinking > or = 3 cups of tea or coffee was associated with elevated risks of spontaneous abortion (adjusted odds ratio = 2.33, 95% CI = 0.92-5.85; and adjusted odds ratio = 2.63, 95% CI = 1.29-5.34, respectively). These results, if replicated, suggest that some ingredient (or correlate) of tea or coffee may account for some of the observed association of caffeine with spontaneous abortion. In this study, caffeine consumption is more strongly related to spontaneous abortion than alcohol or cigarette use in early pregnancy. PMID- 8728438 TI - Diet and rheumatoid arthritis in women: a possible protective effect of fish consumption. AB - Some researchers have hypothesized that omega-3 fatty acids, found primarily in fish oils, may protect against the development of rheumatoid arthritis. We conducted a population-based case-control study in women, comparing 324 incident rheumatoid arthritis cases with 1,245 controls. We used a food frequency questionnaire to ascertain diet during a 1-year period 5 years before a reference date (first physician visit for joint-symptoms). Consumption of broiled or baked fish, but not of other types of fish, was associated with a decreased risk of rheumatoid arthritis. The adjusted odds ratios (OR) for 1- < 2 servings and > or = 2 servings of broiled or baked fish per week, compared with < 1 serving, were 0.78 [95% confidence interval (CI) = 0.53-1.14] and 0.57 (95% CI = 0.35-0.93). Other analyses showed associations with protein as a percentage of calories (adjusted OR for the top quartile as compared with the bottom quartile = 0.65; 95% CI = 0.46-0.94) and total calories (adjusted OR for the top quartile = 1.62; 95% CI = 1.15-2.28). The associations with broiled or baked fish, protein, and calories became stronger when we restricted our analysis to cases positive for rheumatoid factor. These results support the hypothesis that omega-3 fatty acids may help prevent rheumatoid arthritis. PMID- 8728439 TI - Association of diet and colorectal adenomatous polyps: dietary fiber, calcium, and total fat. AB - We conducted a case-control study to investigate the relation between dietary fiber, calcium, and total fat and the risk of colorectal adenomatous polyps. We used a food frequency questionnaire to assess the usual diet for 157 cases and 480 controls. In multivariate analyses, dietary fiber was inversely associated with risk of adenomatous polyps. The odds ratio (OR) for individuals in the highest vs the lowest quartile was 0.5 [95% confidence interval (CI) = 0.3-0.9]. We found an inverse association between dietary calcium and risk of adenomatous polyps, but the protective effect was present only for individuals in the fourth vs the first quartile (OR = 0.7; 95% CI = 0.3-1.3). Intake of total fat was positively associated with risk of adenomatous polyps, but we saw no consistent trend. Calcium intake appeared to modify the effect of total fat intake on the risk of adenomatous polyps. PMID- 8728440 TI - Maternal occupation and pregnancy outcome. AB - Few studies have addressed the effect of maternal employment on late pregnancy outcomes. The National Maternal and Infant Health Survey, a probability sample of U.S. livebirths, stillbirths, and infant deaths in 1988, provided an opportunity to evaluate mothers' jobs in relation to preterm delivery, very low birthweight ( < 1,500 gm), moderately low birthweight (1,500-2,499 gm), small-for-gestational age (SGA) birth, stillbirth, and infant death. We aggregated mothers' jobs, which were ascertained by mailed questionnaire or telephone interview, into categories for analysis. We considered jobs held at any time during pregnancy and jobs held during the fifth month of pregnancy. Relative to the referent group of clerks, textile workers had adjusted odds ratios of 1.5 or greater for all outcomes, with elevated risks also found sporadically for food service workers (preterm delivery, SGA birth, stillbirth) and electrical equipment operators (all outcomes except for still-birth and infant death). Janitors had elevated adjusted odds ratios of 2.0 or greater for preterm delivery and stillbirth. Relative to clerks, teachers and librarians tended to have reduced risks for adverse outcomes. PMID- 8728441 TI - Alcohol and smoking in relation to the prevalence of adenomatous colorectal polyps detected at sigmoidoscopy. AB - Cigarette smoking has been associated with adenomatous polyps of the large bowel but not with increased risk of colorectal cancer. Giovannucci et al recently proposed a hypothesis to explain this inconsistency. A key testable aspect of the hypothesis is that smoking in the distant past increases the risk of large polyps. Questions also remain about the association between colorectal polyps and consumption of alcohol. To address these issues, we examined data from 488 cases with adenomatous polyps and 488 controls. Subjects were members of a prepaid health plan in Los Angeles who had a sigmoidoscopy in 1991-1993. As expected, the adjusted odds of polyps in current smokers compared with never-smokers was increased [odds ratio = 2.43; 95% confidence interval (CI) = 1.56-3.79]. For those who had smoked in the distant past (for example, 30 or more pack-years before 20 years ago), the adjusted odds of an adenoma > or = 1 cm, relative to nonsmokers, was 0.88 (95% CI = 0.23-3.42). The adjusted odds of polyps in those consuming > or = 46 gm per day of alcohol compared with nondrinkers was 1.50 (95% CI = 0.72-3.13). Although imprecise, these data do not support the hypothesis that past smoking increases the risk of large polyps, but our results indicate a weak association between alcohol use and risk of adenomatous polyps. PMID- 8728442 TI - Silica and aluminum in drinking water and cognitive impairment in the elderly. AB - We studied the relation between silica and aluminum levels in drinking water and the risk of cognitive impairment using data from a population-based survey of 3,777 French subjects age 65 years and older. We also studied the effect of pH and the concentrations of calcium, magnesium, fluorine, zinc, copper, and iron. We used a mixed effects logistic regression adjusting for age, sex, educational level, and occupation of the subjects. We confirmed the inverse relation previously found between calcium level and cognitive impairment. We found no important association between cognitive impairment and fluorine, magnesium, iron, copper, or zinc. The association between cognitive impairment and aluminum depended on the pH and the concentration of silica: high levels of aluminum appeared to have a deleterious effect when the silica concentration was low, but there was a protective effect when the pH and the silica level were high. The threshold for an aluminum effect, however, was very low (3.5 micrograms per liter) and did not support the hypothesis of a deleterious effect for only high levels of aluminum. PMID- 8728443 TI - Confidence intervals for measures of interaction. AB - Interaction, defined as departure of disease rates from an additive model, can be measured by the relative excess risk due to interaction, or the attributable proportion due to interaction. Point estimates can be obtained using multiple logistic regression. Using simulated case-control data, we compare several confidence interval estimation techniques for these measures. These include a symmetrical interval based on the delta method estimate of the variance, and three types of bootstrap confidence intervals. One such bootstrap method has coverage closest to the nominal level and is the most evenly balanced with respect to the direction in which intervals miss the true value. The estimation methods are applied to data from an actual case-control study, and the results are interpreted in light of the simulation study. PMID- 8728444 TI - Assessment of cellular telephone and other radio frequency exposure for epidemiologic research. AB - Epidemiologists are now embarking on the evaluation of the hypothesis that exposure to radio frequency energy from low-power wireless communication devices, such as portable cellular telephones, causes brain cancer and other adverse health outcomes. Even in the laboratory, exposures from radio frequency sources are difficult to quantify; their measurement in large populations for epidemiologic study is challenging. In this paper, we outline the nature and magnitude of these exposures and discuss the prospects for obtaining useful measures of exposure for epidemiologic research. PMID- 8728445 TI - Utility of telephone company records for epidemiologic studies of cellular telephones. AB - We conducted a survey of over 5,000 telephone users who were customers of one large cellular telephone company covering four major geographical areas. Our primary goal was to assess the utility of ascertaining information on telephone use and type from telephone company records. We compared information from 3,949 respondents with corresponding data from company billing records. We found that 48% of the account holders were sole users, and 69% were the primary user, meaning that they accounted for at least 75% of the use. Respondent reports of amount of telephone use were highly correlated with data on the billing record (r = 0.74). Respondent reports of telephone type were similarly correlated with data from the manufacturer (r = 0.92). We also inquired about telephone holding patterns, since these have implications for exposure. Most users reported favoring one side of the head when using the telephone, but the side of the head used was not strongly associated with handedness. PMID- 8728446 TI - Overall mortality of cellular telephone customers. AB - Unlike mobile cellular telephones, in which the antenna is not part of the handset, a portable cellular telephone exposes the user's head to radio frequency energy transmitted from the antenna. This exposure has prompted concerns about potential biological effects, including brain cancer. As a first step in a record based mortality surveillance of cellular telephone customers, we report on overall mortality of a cohort of more than 250,000 portable and mobile telephone customers during 1994. We found age-specific rates to be similar for users of the two types of telephones. For customers with accounts at least 3 years old, the ratio of mortality rates in 1994 for portable telephone users, compared with mobile telephone users, was 0.86 (90% confidence interval = 0.47-1.53). PMID- 8728447 TI - Risk factors for pregnancy-induced hypertension in women at high risk for the condition. Italian Study of Aspirin in Pregnancy Group. AB - To analyze determinants of the risk of pregnancy-induced hypertension (PIH) with or without proteinuria, we compared characteristics of women enrolled in the Italian Study of Aspirin in Pregnancy who developed PIH and those who did not. A total of 756 women were included in the present analysis; of these, 132 women (17%) developed PIH during the trial. The risk of developing PIH tended to increase with maternal age: in comparison with women age 20-25 years, the odds ratio (OR) estimates of risk ratio were 3.5 [95% confidence interval (CI) = 1.6 7.1] in women age 26-30 years and 4.2 (95% CI = 1.9-8.8) in those age > 30 years. There was little relation between development of PIH and education. PIH risk increased according to nonpregnant body mass index; in comparison with women with Quetelet's index (kg per m2) < 25, the OR estimates were 1.7 (95% CI = 1.1-2.7) and 2.1 (95% CI = 1.3-3.6), respectively, for women with a value for Quetelet's index of > 25-30 and > 30. Parous women were at decreased risk of PIH: in comparison with nulliparas, the ORs were 0.7 (95% CI = 0.4-1.0) and 0.5 (95% CI = 0.3-0.9), respectively, in women reporting 1 or > or = 2 births. There was no important relation between previous spontaneous or induced abortion and PIH risk. PMID- 8728448 TI - Indices of exposure to fetal and sperm antigens in relation to the occurrence of breast cancer. AB - We used data from a population-based case-control study to evaluate Janerich's hypothesis that reduced risk of breast cancer in women with multiple marriages may be attributable to an immune response to fetal or sperm antigens. Risk of breast cancer in women with multiple marriages was reduced relative to that in women who had been married only once; however, there was no indication that such risk was reduced among women whose full-term pregnancies were fathered by different men, relative to women whose pregnancies were each fathered by the same man. Increasing lifetime number of male sexual partners was associated with a trend of decreasing risk of breast cancer. Our results indicate that, if there are effects of exposure to fetal or male antigens on risk of female breast cancer, their impact may be heterogeneous. PMID- 8728449 TI - Reliability of information on cigarette smoking and beverage consumption provided by hospital controls. AB - We compared data from interviews of patients admitted to hospital and interviews repeated at home for 400 hospital controls. Of these, 294 (73.5%) could be re interviewed (responders). Never smoking was reported by 44% of subjects in the in hospital interviews and by 40% in the repeat at-home interviews. Among those interviewed in hospital, 38% claimed to be nondrinkers, as did 38% of responders, but only 28% characterized themselves as nondrinkers in the repeat interview. For consumption of regular coffee, 15% reported none in the selected sample, 14% among responders, but only 6% in the repeat at-home interview. Agreement was good for decaffeinated coffee and tea consumption. PMID- 8728450 TI - Differences between spontaneous and induced abortions as risk factors for breast cancer. PMID- 8728451 TI - Healthy screened bias in epidemiologic studies of cancer incidence. PMID- 8728452 TI - The decline of SIDS: a success story for epidemiology. PMID- 8728453 TI - Mechanistic reasoning in sociobehavioral epidemiology: the influence of race and ethnicity on cancer survival. PMID- 8728454 TI - Different, not biased. PMID- 8728455 TI - Parental age and breast cancer mortality. PMID- 8728456 TI - Software for optimal matching in observational studies. PMID- 8728457 TI - The black box revisited. PMID- 8728458 TI - The validity of the extension-rotation test as a clinical screening procedure before neck manipulation: a secondary analysis. AB - OBJECTIVE: To determine the validity of the neck extension-rotation test as a clinical screening procedure to detect decreased vertebrobasilar blood flow that might be associated with dizziness. DESIGN: Secondary analysis of a clinical screening test. METHODS: Twelve subjects with dizziness reproduced by the extension-rotation test and 30 healthy control subjects had Doppler ultrasonography examination of their vertebral arteries with the neck extended and rotated. Vascular impedance to blood flow was measured and the presence of signs and symptoms of vertebrobasilar ischemia was recorded. RESULTS: Cut-off points for validity estimates were derived through the percentile and Gaussian methods using impedance to blood flow as the standard. The sensitivity of the extension-rotation test for increased impedance to blood flow was 0%, regardless of the selected cut-off point. The specificity rates for the left vertebral artery were 71% and 67% for the percentile and Gaussian methods, respectively. The extension-rotation test was more specific on the right side, with a rate varying from 90% with the percentile method to 86% with the Gaussian technique. The positive predictive value of the test was 0% and its negative predictive value ranged from 63% to 97%. CONCLUSION: We were unable to demonstrate that the extension-rotation test is a valid clinical screening procedure to detect decreased blood flow in the vertebral artery. The value of this test for screening patients at risk of stroke after cervical manipulation is questionable. PMID- 8728459 TI - Lasting changes in passive range motion after spinal manipulation: a randomized, blind, controlled trial. AB - PURPOSE: To study whether a 3-wk series of spinal manipulation has any lasting effect on passive cervical range of motion. DESIGN: Randomized, controlled trial with a blind observer. SETTING: Ambulatory outpatient facility in an independent National Health Service funded chiropractic research institution. PARTICIPANTS: Thirty-nine headache sufferers who, on entering the study, displayed objectively decreased passive cervical range of motion. These subjects were recruited from 400 headache sufferers who responded to newspaper advertisements. INTERVENTION: Half of the group received high-velocity, low-amplitude cervical manipulation twice a week for 3 wk. The other half received low-level laser in the upper cervical region and deep friction massage in the lower cervical/upper thoracic region, also twice a week for 3 wk. MAIN OUTCOME MEASURE: Goniometrically assessed passive range of motion of the cervical spine. RESULTS: Although passive cervical range of motion increased in both groups during the trial period, there were no statistically significant differences between the two groups 1 wk after the last treatment. CONCLUSION: It seems that any changes in passive range of motion after spinal manipulation are of a temporary nature. PMID- 8728460 TI - Ear infection: a retrospective study examining improvement from chiropractic care and analyzing for influencing factors. AB - OBJECTIVE: The aims of this study were to determine (a) if the patients improved while under chiropractic care; (b) how many treatments were needed to reach improvement; and (c) which factors were associated with early improvement. DESIGN: Cohort, nonrandomized retrospective study. SETTING: Private chiropractic practice in a Minneapolis suburb. PARTICIPANTS: Forty-six children aged 5 yr and under. INTERVENTION: All treatments were done by a single chiropractor, who adjusted the subluxations found and paid particular attention to the cervical vertebrae and occiput. Sacral Occipital Technique-style pelvic blocking and the doctor's own modified applied kinesiology were also used. Typical treatment regimen was three treatments per week for 1 wk, then two treatments per week for 1 wk, then one treatment per week. However, treatment regimen was terminated when there was improvement. OUTCOME MEASURE: Improvement was based on parental decision (they stated that the child had no fever, no signs of ear pain, and was totally asymptomatic), and/or the child seemed to be asymptomatic to the treating DC and/or the parent stated that the child's MD judged the child to be improved. A data abstraction form was used to determine number of treatments used and presence of factors possibly associated with early improvement. RESULTS: 93% of all episodes improved, 75% in 10 days or fewer and 43% with only one or two treatments. Young age, no history on antibiotic use, initial episode (vs. recurrent) and designation of an episode as discomfort rather than ear infection were factors associated with improvement with the fewest treatments. CONCLUSION: Although there were several limitations to this study (mostly because of its retrospection but also, significantly, because very little data was found regarding the natural course of ear infections), this study's data indicate that limitation of medical intervention and the addition of chiropractic care may decrease the symptoms of ear infection in young children. PMID- 8728461 TI - The progressive making of a foremost chiropractic researcher: Fred Illi from Geneva. AB - Until after the workshop on the research status of Spinal Manipulative Therapy at the National Institute of Neurological and Communicating Disorders and Strokes (NINCDS) in Bethesda, Maryland, in 1975, research had been a very elusive yet critical part of chiropractic evolution. In the past, chiropractic education emphasized basic sciences for legitimacy. To increase their competitive edge, chiropractors all too frequently looked to medical research rather than conducting research of their own. A few exceptionally gifted individuals and agencies within the profession developed new paradigms, improved modalities, and clinical mastery. Very little descriptive information has been made available to assess either these individuals or their achievements. With more than half the profession having less than 10 yr of practice, it becomes necessary to kindle a reappraisal of what once was in the particular area of chiropractic research. The case of Fred Illi has been chosen to illustrate the inherent difficulties that faced anyone bold enough to try and coherent enough to be successful. PMID- 8728462 TI - Upper cervical instability: fact or fiction? AB - OBJECTIVES: The purpose of this review is to examine the internal validity of the standardized clinical stability tests for the upper cervical spine in relation to symptomatology. Whether radiology can confirm the clinical diagnosis is also examined. The importance of radiology in this situation, the pathogenesis and the prevalence of atlanto-axial hypermobility and the clinical symptomatology are discussed. DATA SOURCES: A literature search from January 1984 to March 1995. We consulted the CD-ROM Medline with the keywords "atlanto-axial instability," "atlanto-axial dislocation," "hypermobility," "cervical spine" and "atlanto-axial joint." Ninety-six Dutch, French, German, and English publications were selected. The Documentation Centre of the Institute for Research and Postgraduate Education Physiotherapy (SWSF) was consulted with the keywords: atlanto-axial joint, upper cervical spine, segmental examination, interobserver-reliability, intraobserver reliability, interobserver variation, intraobserver variation, manual therapy, examination, diagnostics. Finally, recent developments and views published during this study were added. RESULTS: There seems to be no correlation between the amount of hypermobility or subluxation and the presence of clinical signs or neurological signs. The clinical signs can vary from relatively diffuse complaints, no symptoms and signs to serious ones. Radiology does not seem to be a reliable diagnostic mechanism in relation to upper-cervical instability. Conventional X-rays fail to give adequate information about atlanto-axial stability. CT-scan and MRI can visualize much more because of the direct sagittal projection but neither is an absolute standard. Furthermore, in relation to upper cervical hypermobility, the validity of radiology is under debate. CONCLUSION: There is no correlation between the measure of hypermobility and the presence of clinical symptoms. Also, the validity of the upper-cervical stability tests is questionable. In diagnostics, every radiological examination measures anatomical and morphological variables, not functional variables. Despite this, CT and MRI should be preferred in diagnostics over conventional functional radiology. PMID- 8728463 TI - Scheuermann's juvenile kyphosis. AB - OBJECTIVE: Two cases are discussed to illustrate two different presentations, progressions and treatments of Scheuermann's juvenile kyphosis. CLINICAL FEATURES: In one case, a 13-yr-old boy suffered from a 2-yr history of lower back pain. Radiographs demonstrated irregularity of the upper lumbar vertebral endplates, associated with Schmorl's nodes. The second case is one of a 14-yr-old boy who was seen in an orthopedic outpatient clinic. Radiographs revealed wedging of the anterior border of T6, T7, and T8 vertebrae with a thoracic spine kyphotic deformity measuring 72 degrees. INTERVENTION AND OUTCOME: The first case was treated conservatively. The patient maintained his improvement at 6 month follow up. The second case was initially treated with a brace that the patient did not wear regularly as directed. The kyphotic deformity progressed from 72 degrees to 92 degrees. An operation was performed to reduce the kyphotic curve and prevent further progression. On review 6 yr later, the patient was well without back pain or other complications. The kyphotic curve measured 65 degrees. CONCLUSION: Scheuermann's juvenile kyphosis is a common spinal deformity in the adolescent. A radiographic appearance of wedging of the anterior portion of the vertebral bodies with marked kyphotic deformity suggests the diagnosis of classical Scheuermann's disease. However, the lumbar type of Scheuermann's disease should be considered in young patient with radiographic evidence of irregular vertebral endplates, Schmorl's nodes and a decreased disc space without wedging. Nevertheless, significant progression of the curve in both the typical and atypical types of Scheuermann's disease is rare, but can occur. An algorithm is presented to facilitate decision making in the management of Scheuermann's juvenile kyphosis. PMID- 8728464 TI - A holistic approach to severe headache symptoms in a patient unresponsive to regional manual therapy. AB - OBJECTIVE: To discuss the case of a patient suffering from severe headache complaints who was previously unresponsive to regional cervical spine care. CLINICAL FEATURES: A prodrome was described. In addition to headache itself, associated symptoms included light sensitivity, blurred vision and nausea. A visit to a hospital emergency room was often necessary to reduce pain intensity. INTERVENTION AND OUTCOMES: Manual chiropractic spinal adjustment were performed as the only treatment intervention. After a course of treatment, the patient reported no visits to the emergency room, even after a 1-yr follow-up, and the average visual analogue pain scores decreased. CONCLUSION: This patient seemed to respond favorably to conservative care that included regions of spine not traditionally associated with headache pain. This suggests that some individuals may require a more comprehensive evaluation if regional care fails to promote a positive response within a few weeks. Controlled, randomized trials will assist in comparing effectiveness of various treatment interventions. PMID- 8728465 TI - A systems model of health care: a proposal. PMID- 8728466 TI - On sounds and reflexes. PMID- 8728467 TI - "Research" and "science" in the first half of the chiropractic century. PMID- 8728468 TI - A risk assessment of spinal manipulation vs. NSAIDs for the treatment of neck pain. PMID- 8728469 TI - How to evaluate intraexaminer reliability from an interexaminer reliability study design. PMID- 8728470 TI - Increased susceptibility of brain slices from carbonic anhydrase II-deficient mice to low [Mg2+]O-induced seizures. AB - Brain pH is thought to be an influential factor in determining susceptibility to seizures. We compared the susceptibility of brain slices from carbonic anhydrase II (CA II)-deficient mice to epileptiform activity induced by low extracellular [Mg2+], with slices from normal littermates, both bathed in artificial cerebrospinal fluid at pH 7.3. In both entorhinal cortex and hippocampal field CA1, epileptiform activity started earlier in CA II-deficient slices. Raising extracellular [CO2] (20%; extracellular pH, 6.7) reversibly blocked the epileptiform activity in normal, but not in CA II-deficient, slices. The data, combined with previous in vivo findings showing an increased resistance of mutants to seizures, suggest the presence of in vivo anticonvulsant acidosis with long-term compensatory changes that lead to in vitro 'proconvulsant' behavior in CA II-deficient slices clamped at pH 7.3. PMID- 8728471 TI - Contribution of vagal afferents to the volume-timing response to expiratory loads in neonatal lambs. AB - Expiratory loads elicit volume-dependent reflex increases in expiratory duration (TE) in adult animals. This reflex is mediated by pulmonary vagal afferents. It was hypothesized that this same reflex in newborn lambs is mediated by vagal afferents. Single-breath graded expiratory loads were presented to 4 day old, anesthetized neonatal lambs through a facemask (FM), via tracheostomy (TR) and after the cervical vagi were severed bilaterally in the TR animals (VAG). In FM and TR studies, expired volume (VE) decreased and TE increased with increasing load magnitude. There was no significant difference in the VE-TE curves for FM and TR. These results demonstrate that pulmonary vagal afferents are mediating the volume-related reflex modulation of TE during expiratory loading in the anesthetized neonatal lamb which is unaffected by bypass (TR) of upper airway afferents. PMID- 8728472 TI - Inhibition of tyrosine phosphorylation attenuates amino acid neurotransmitter release from the ischemic/reperfused rat cerebral cortex. AB - Protein tyrosine phosphorylation plays an important role in neuronal function. In this study we have examined the effects of inhibition of tyrosine phosphorylation on the extracellular levels of four neurotransmitter amino acids (aspartate, glutamate, gamma-aminobutyric acid (GABA) and glycine) and of the non-transmitter amino acid phosphoethanolamine during cerebral ischemia and reperfusion in a rat four vessel occlusion model. In comparison with the control group, the tyrosine kinase inhibitor genistein significantly depressed ischemia/reperfusion-evoked efflux of these amino acids, with the exception of GABA, into cerebral cortical superfusates. GABA efflux was non-significantly reduced. These results suggest that tyrosine phosphorylation is involved in the ischemia-evoked efflux of amino acids into the extracellular milieu, likely as a consequence of the phosphorylation of microtubule-associated protein kinase (MAP kinase) and downstream activation of PLA2 in the plasma membrane. Amino acid efflux would occur, in part, as a consequence of the ensuing disruption of plasma membrane integrity and leakage of cytoplasmic constituents along their concentration gradients. PMID- 8728473 TI - Tetrahydroberberine suppresses dopamine-induced potassium current in acutely dissociated CA1 pyramidal neurons from rat hippocampus. AB - Effects of tetrahydroberberine (THB) on dopamine (DA)-induced response have been investigated in single pyramidal neurons freshly dissociated from CA1 area of the hippocampus using the nystatin perforated patch-clamp, whole-cell recording technique under voltage-clamp configuration. At a holding potential (VH) of 20mV, DA-induced responses included a transient outward current, a slow inward current and a combination of the two. The outward current had a reversal potential of -83.5 +/- 8.0 mV which was close to K+ equilibrium potential and was sensitive to TEA, suggesting that this outward current was carried by K+. Application of THB reversibly suppressed three type responses induced by DA with different degrees of inhibitory ratio. THB inhibited the DA-induced outward current in a concentration-dependent manner with an IC50 of 13 microM. The maximal value of the concentration-response curve for DA-induced outward current was suppressed by THB, suggesting a non-competitive inhibition. The results support the hypothesis that THB acts as a novel dopaminergic system antagonist. Furthermore, THB inhibits the DA-induced K+ current non-competitively in single dissociated cells, implying that THB may exhibit other pharmacological action on the central nervous system. PMID- 8728474 TI - Electron microscopic demonstration of tyrosine hydroxylase-immunoreactive interplexiform cells in the lamprey retina. AB - To clarify the controversies about the existence (or not) of dopaminergic interplexiform cells in the lamprey retina, we have performed an immunocytochemical electron microscopic study of the retina of the river lamprey Lampetra fluviatilis, using anti-tyrosine hydroxylase antibody. We demonstrate the presence of immunoreactive processes in both inner (IPL) and outer (OPL) plexiform layers. The external processes are in close contact with horizontal cell processes and photoreceptor terminals in the OPL, but do not make classical synapses. Some of them are ensheathed within Muller cell cytoplasm. Thus, dopaminergic interplexiform cells do exist in Lampetra fluviatilis, but the absence of synapses rather supports a paracrine action of dopamine in the outer retina. PMID- 8728475 TI - C-Fos proteins are not involved in the activation of preproenkephalin gene expression in rat brain by peripheral electric stimulation (electroacupuncture). AB - The present work was designed to study the role of the oncogene product c-Fos in activating the transcription of preproenkephalin (PPE) gene following a kind of peripheral electric stimulation known as electroacupuncture (EA) stimulation. The temporal patterns of rat brain c-fos and PPE mRNA expression were evaluated using the method of Northern blotting, showing that c-fos mRNA expression, which peaked at 2 h after the termination of EA, was always ahead of the PPE mRNA expression which began at 4 h and peaked at 48 h after EA. The methods of immunocytochemistry (ICC) and in situ hybridization (ISH) techniques were combined to identify the co-existence of c-Fos protein and PPE mRNA at the cellular level. The results showed that only a small percentage of PPE mRNA containing neurons depicts Fos-like immunoreactive nuclei. These findings suggest that c-Fos protein may not be involved in the activation of brain PPE gene transcription induced by peripheral electric stimulation. PMID- 8728476 TI - Acoustic modulation of electrically evoked distortion product otoacoustic emissions in gerbil cochlea. AB - In order to study the linearity of outer hair cell fast electromotility in vivo, an acoustic tone was used to interact with the electrically evoked distortion product otoacoustic emissions. Otoacoustic emissions at the primary frequencies (f1, f2, where f1 < f2) and the distortion frequencies (2f1 - f2 and f2 - f1) were evoked by a complex current, with f1 and f2 components, delivered to the gerbil round window. An externally given acoustic tone at the frequency f1 or f2 with appropriate phase and level can completely abolish both the 2f1 - f2 and f2 f1 distortion tones. Because the external tone causes basilar membrane vibration at its natural topographic locations, this result indicates that the observed distortion tones were generated near the locations of f1 and f2 frequencies on the basilar membrane and that no distortion occurred from the stimulated cells near the electrode. The study strongly suggests a linear electromechanical transduction of the outer hair cells in the sensitive cochlea. PMID- 8728477 TI - Relationships between beta-amyloid (A beta) deposits and blood vessels in patients with sporadic and familial Alzheimer's disease. AB - The density of diffuse, primitive and classic beta-amyloid (A beta) deposits was studied in relation to the incidence of blood vessels in the superior frontal gyrus of nine cases of sporadic Alzheimer's disease (SAD), two cases of familial Alzheimer's disease (FAD) with amyloid precursor protein (APP) mutations (APP717, Val --> Ile), and eight cases of FAD not linked to chromosomes 21, 14 or 1. Stepwise multiple regression was used to determine for each patient whether variations in the density of A beta deposits along the cortex were significantly correlated with the incidence of blood vessels. In the majority of FAD and SAD cases, the density of the diffuse and primitive type A beta deposits was not related to blood vessels. However, the incidence of the larger diameter (> 10 microns) blood vessels was positively correlated with the density of the classic A beta deposits in eight (89%) SAD and two (20%) FAD cases. The data suggest that the densities of vessels and deposits were not significantly correlated between cases but only within cases, suggesting a strictly local effect. In addition, the spatial association between classic A beta deposits and blood vessels may be more apparent in SAD compared with FAD cases. PMID- 8728478 TI - Analgesic effects of fibroblast growth factor in the rat. AB - Intraperitoneal (i.p.) injection of acidic fibroblast growth factor (aFGF) to Sprague-Dawley rats induced short-lasting analgesia as measured by tail-flick latency (TFL) test. The maximum effect, a 26% increase in tail-flick latency, was obtained 15 min following 1 microgram i.p. aFGF. By 30 min the effect was considerably reduced, and was no longer present by 45 min after treatment. Administration of heat-inactivated aFGF or a hybrid form of aFGF (CLYT/aFGF) that, although active, is unable to cross the blood-brain barrier (BBB), caused no analgesia. Furthermore, the analgesic effects of aFGF were prevented by pretreatment with the nitric oxide synthase inhibitor, L-NG-nitroarginine methyl ester (L-NAME). Our findings demonstrate an analgesic effect of FGF, which requires crossing of BBB and implicates the nitric oxide pathway. PMID- 8728479 TI - The dynamic response of feline Golgi tendon organs during recovery from nerve injury. AB - The dynamic response of tendon organs to isometric contractions of activating motor units has been examined during recovery from nerve crush or nerve transection followed by suture repair. After nerve crush the characteristic response was rapidly restored, although the early and late phases of the dynamic response were altered differentially. Following nerve transection, recovery was much poorer and many responses were abnormal. Normal responses were only observed in a minority of tendon organ-motor unit interactions but every tendon organ studied did respond normally to at least one motor unit, with a range of dynamic sensitivities similar to normal. This suggests that the abnormalities observed reflect changes in the mechanical input to the organ, due to motor unit reorganisation, rather than abnormalities of the transduction process. PMID- 8728480 TI - Specific up-regulation of the POU domain transcription factor Oct-2 following axotomy. AB - Peripheral nerve damage causes a dramatic alteration to the gene expression in primary sensory neurons, changes within the neuronal cell body giving rise to an altered phenotype, adapted for axonal regeneration. Such changes suggest an alteration in activity, or levels, of cellular transcription factors. The POU family transcription factor Oct-2 is known to be induced in sensory neurons by nerve growth factor (NGF) and might therefore be affected by the removal of target-derived NGF following axotomy. Paradoxically, however, the expression of Oct-2 showed a transient increase of two- to three-fold 24 h after axotomy. In contrast, axotomy had no effect on the levels of the Brn-3 sub-family of POU proteins, indicating that this effect was specific for Oct-2. PMID- 8728481 TI - The effect of pituitary adenylate cyclase activating peptide (PACAP) on the nociceptive formalin test. AB - Recent studies have suggested that pituitary adenylate cyclase activating peptide (PACAP) may be involved in nociceptive transmission. The present study examined the effect of low-dose PACAP-27 on nociceptive behavior using the formalin test. PACAP-27 was administered intrathecally. Twenty minutes later, formalin (50 microliters, 5%) was injected subcutaneously into the dorsal surface of the right hind paw. Intrathecal PACAP-27 at 0.6 pmol suppressed the second phase response to formalin, while 5 pmol depressed both phases. PACAP-27 at 5 pmol did not impair motor function. Hence, the data suggest that the effect of PACAP-27 on formalin-induced pain-related behavior is specific. The findings may lead to a better understanding of the role of PACAP in nociceptive transmission. PMID- 8728482 TI - MDL 12330A inhibits the non-neuronal adenylyl cyclase from the freshwater snail Planorbarius corneus, but the neuronal enzyme is activated by this compound. AB - N-(Cis-2-phenyl-cyclopentyl)azacyclotridecan-2-imine-hydrochloride (MDL 12330A), considered an inhibitor of adenylyl cyclase, has been tested on the enzyme activity of neuronal and non-neuronal tissues from the freshwater snail Planorbarius corneus. The drug dose-dependently activates the basal as well as agonist-stimulated adenylyl cyclase in the ganglionic preparations, while it exerts an inhibitory effect on the enzyme present in the non-nervous tissues examined. SQ 22536 and forskolin, respectively an inhibitor and activator of adenylyl cyclase, behave as generally reported both in central and peripheral tissues of the snail. This is, to our knowledge, the first report of a stimulatory action of MDL 12330A on an adenylyl cyclase system. PMID- 8728483 TI - Presence of the voltage-gated potassium channels sensitive to charybdotoxin in inhibitory presynaptic terminals of cultured rat hippocampal neurons. AB - To determine whether the charybdotoxin-sensitive subtypes of voltage-gated K+ channels (Kv1.2 and Kv1.3) exist in inhibitory pre-synaptic terminals, effects of K+ channel blockers including TEA, charybdotoxin (ChTX), iberiotoxin (IbTX), kaliotoxin (KTX) and margatoxin (MgTX) on the inhibitory transmission were examined with cultured rat hippocampal neurons. Monosynaptic inhibitory postsynaptic currents (IPSCs) evoked by electrical stimulation of single presynaptic neurons were recorded from the whole-cell clamped postsynaptic neurons. In the presence of TEA, application of ChTX greatly increased the amplitude of IPSCs. A specific maxi-K+ channel blocker IbTX failed to augment IPSCs. KTX and MgTX, both of which block Kv1.3 but not Kv1.2, mimicked the facilitating effect of ChTX. In the absence of TEA, application of ChTX increased the IPSC amplitude significantly, while IbTX was without effect. These results indicate that the ChTX-sensitive subtypes of voltage-gated K+ channels, most likely Kv1.3, contribute to the repolarization of action potentials at presynaptic terminals of hippocampal inhibitory neurons, and that the ChTX induced facilitation of the transmission can be explained by its effects on the Kv channels rather than maxi-K+ channels. PMID- 8728484 TI - Presynaptic localization of a metabotropic glutamate receptor, mGluR4a, in the cerebellar cortex: a light and electron microscope study in the rat. AB - Localization of a metabotropic glutamate receptor, mGluR4a, was immunohistochemically examined in the rat cerebellum with an antibody, which was produced by using a synthetic peptide corresponding to a C-terminal sequence of rat mGluR4a. Marked mGluR4a-like immunoreactivity (mGluR4a-LI) was seen in neuropil of the molecular layer of the cerebellar cortex. Electron microscopically, mGluR4a-LI was observed in many axon terminals in the molecular layer. These axon terminals showing mGluR4a-LI were filled with round synaptic vesicles and were in asymmetric synaptic contacts most frequently with dendritic spines. The results indicate that mGluR4a are located presynaptically in the parallel fibers arising from the granule cells in the cerebellar cortex. PMID- 8728485 TI - A neural influence on the appearance of sodium current in Xenopus embryonic myocytes. AB - Regulation of early embryonic expression of sodium current in cultured Xenopus laevis myocytes was investigated. In myocytes isolated before innervation and cultured for about 1 day, only 51% expressed sodium current at a mean density of 74 +/- 20 pA/pF (mean +/- SEM; n = 26), inadequately reflecting the functional development of skeletal muscle in vivo at this time. This cell-autonomous expression pattern could be modulated. First, co-culture of myocytes and dissociated neural tissue induced additional sodium current expression (82%; 172 +/- 31 pA/pF, n = 14) and, secondly, removal of calcium from the culture medium reduced sodium current density (13 +/- 6 pA/pF; n = 5). The former results suggest that a diffusible factor released from differentiating neurons is able to initiate the expression of sodium channels in myocytes. PMID- 8728486 TI - Significantly increased frequency of the apolipoprotein E epsilon 4 allele in elderly non-demented leprosy patients. AB - Apolipoprotein E (apo E) genotypes in 350 leprosy patients were examined and compared with those of 870 age-matched controls. The allelic frequencies of the apo E gene did not differ between demented patients with leprosy and controls. However, the frequency of apo E epsilon 4 allele was significantly higher in non demented leprosy patients than in controls (P < 0.001). Of special interest is that the prevalence of E3/4 genotype in non-demented leprosy patients increased significantly with age, being 14.1%, 24.4%, and 28.3% in the 60s, 70s, and 80s, respectively (P < 0.05). These data suggest that apo E epsilon 4 is not a risk factor for senile dementia in elderly leprosy patients, and there exist factors to overcome the risk of apo E4 in leprosy patients. PMID- 8728487 TI - Daytime melatonin administration enhances sleepiness and theta/alpha activity in the waking EEG. AB - It is still controversial whether the pineal hormone melatonin can be characterized as a hypnotic. We therefore measured subjective sleepiness and waking EEG power density in the range of 0.25-20 Hz after a single dose of melatonin (5 mg). During an 8 h mini-constant routine protocol, melatonin administered in a double blind cross-over design to healthy young men at 1300 h or 1800 h increased subjective sleepiness, as rated half-hourly on three different scales (Visual Analogue Scale, Akerstedt Sleepiness Symptoms Check List, Akerstedt Sleepiness Scale) and objective fatigue as evidenced by augmented waking EEG power density in the theta/alpha range (5.25-9 Hz). The increase in subjective sleepiness reached significance 40 min and 90 min after melatonin administration (at 1300 h and 1800 h, respectively) and lasted for 3 h (at 1300 h) and 5 h (at 1800 h). The increase in the theta/alpha frequencies of the waking EEG occurred immediately after melatonin ingestion and stayed significantly higher parallel to the higher sleepiness ratings. However, the EEG changes appeared before the subjective symptoms of sleepiness became manifest. There was a significant correlation between salivary melatonin levels and the timing of increased subjective sleepiness. Melatonin had no effects on mood. PMID- 8728488 TI - Review of external ocular disease associated with aids and HIV infection. AB - External ocular disease associated with human immunodeficiency virus (HIV) infection can often be overlooked by the eye care practitioner. Different types of external ocular disease can be an indication of the patient's overall immune status as well as the stage of HIV infection. The external ocular sequelae of HIV infection can be of visual consequence for the patient. Eye care practitioners need to become familiar with these conditions. The diagnosis and management of the following ocular conditions associated with HIV infection are reviewed: conjunctival microvascular disease, dry eye, allergic conjunctivitis, microsporidial keratoconjunctivitis, herpes zoster ophthalmicus, herpes simplex keratitis, molluscum contagiosum, fungal keratitis, bacterial keratoconjunctivitis, and Kaposi's sarcoma (KS). PMID- 8728489 TI - Vitreous chamber elongation is responsible for myopia development in a young adult. AB - In this case report, we present 3 years of longitudinal, refractive component data for a young adult who became myopic and progressed in myopia (late-onset myopia, LOM) during that period. We found a high correlation between refractive error and axial length (AL). The axial elongation was a consequence of the change in the vitreous chamber depth (VCD). Furthermore, the change in AL and VCD corresponded quantitatively to the change in refractive error. Corneal curvature change was not correlated with refractive error change. These findings support the suggestion that VCD is the refractive component that changes in LOM development. PMID- 8728490 TI - Age-related changes in the accommodation mechanism. AB - Presbyopia is caused by changes in the visco-elastic properties of the lens or the choroid or both. Far-to-near (FN) accommodation dynamics are dominated by the properties of the lens, whereas near-to-far (NF) accommodation dynamics depend on the properties of the lens as well as the choroid. The aim of this study was to gain more insight into the cause of presbyopia by analyzing how FN and NF accommodation dynamics change as a function of age. Changes in axial lens thickness during accommodation were measured in vivo using continuous ultrasonographic biometry. The measurements were analyzed with the use of a biomechanical model of the mechanism of accommodation. Fitting the model to the responses yields time constants of FN and NF accommodation. The time constants FN and NF increase with age at an approximately equal rate: 7 ms/year (+/- 1.8 SEM) and 6 ms/year (+/- 1.6 SEM), respectively. This result supports a lenticular cause of presbyopia. In addition, estimations of the lens damping coefficients at different ages are given for the first time: the damping coefficient of the lens increases 20-fold between 15 and 55 years of age. PMID- 8728491 TI - Routine optometric care in nursing homes: a retrospective study. AB - Residents of nursing homes do not have the same access to health care as do others. A retrospective study of the records of optometric examinations at three nursing homes with demographics presumably similar to those of most nursing homes in the United States was performed in order to determine the prevalence of vision anomalies in this population and to determine if optometric intervention was beneficial. Sixty-five percent of the patients received an immediate benefit from the optometric examination in either treatment or referral to another health care provider for an undetected or poorly controlled health problem. We concluded that this type of service was needed by the patients in the nursing homes and was professionally rewarding to the optometrist providing it. PMID- 8728492 TI - Evaluation of soft contact lens fitting characteristics. AB - The assessment of soft contact lens fit by observation of the lens behavior on the eye is one of the most common procedures undertaken in contact lens practice. Nevertheless, the guidelines for doing so have been derived empirically with little or no research into the usefulness of these observations. Over 2000 evaluations of soft contact lens fit were analyzed to determine the predictive value of various indicators of contact lens fit, namely: subjective comfort, lens centration, postblink movement (PBM), version and upgaze lag, and tightness assessed by the push-up test. The receiver-operator characteristic (ROC) curves were prepared for each variable by plotting sensitivity values against false positive rates (FPRs) for various cut-off points. Sensitivity was defined as the proportion of "bad" fits correctly identified by a given variable; FPR was defined as the proportion of "good" fits incorrectly identified as bad fits. With each fitting variable, the optimum cut-off point and its corresponding sensitivity and FPR were determined from the ROC curves. Subjective comfort was found to be of little value in identifying tight fitting lenses but was of some limited value in identifying loose fitting lenses, particularly high water content (HWC) lenses. The push-up test was the most accurate single test of lens fit acceptability having a high sensitivity for both tight (92%) and loose (80%) lens fits. Assessment of PBM was a sensitive indicator of tight fitting lenses (95%), but was a poor indicator of loose fitting lenses, having a low sensitivity (51%). Assessment of version lag showed better sensitivity than upgaze lag in assessing loose fits, whereas upgaze lag showed better sensitivity for tight lens fits. Lens centration was of no predictive value in identifying tight fitting lenses but, as expected, was of some value in identifying loose fitting lenses. The results of this study enable a critical reassessment of the procedures that practitioners follow when fitting soft contact lenses. PMID- 8728493 TI - Visual predictors of reading performance in kindergarten and first grade children. AB - PURPOSE: A masked investigation of the relation between performance on various vision tests and reading was conducted with 90 kindergartners (mean age 5.73 years) and 91 first graders (mean age 6.76 years) from a middle class, suburban, elementary school near Cleveland, Ohio. METHODS: Vision testing included the Modified Clinical Technique (MCT), +/- 2.00 D flipper lenses with red/green suppression check for accommodative facility, and Randot for stereoacuity. Reading performance was independently evaluated with the Metropolitan Achievement Test 6 Reading Test and teachers' assessments. RESULTS: The results revealed that accommodative facility was predictive of successful reading performance in 7-year olds (p = 0.0431), first graders (p = 0.0125), and in the entire subject group when age (p = 0.0254) or grade (p = 0.0224) was controlled. Failure on the MCT was significantly associated with decreased reading skill in 5-year-olds (p = 0.0431). In addition, stereoacuity worse than 100 sec arc (p = 0.0316), MCT failure plus stereoacuity worse than 50 sec arc (p = 0.0316), and accommodative facility (p 0.0155) were predictive of whether children of average intelligence would show successful or unsuccessful reading ability. CONCLUSIONS: Thus, visual performance was significantly related to reading performance even in children of average intelligence when IQ was partially controlled. Also, the predictive value of the MCT for reading achievement could be improved by the addition of a referral criterion for stereoacuity. This would make the results of MCT screening more readily applicable to educators. PMID- 8728494 TI - Clinical findings before the onset of myopia in youth: 2. Zone of clear single binocular vision. AB - BACKGROUND: Many theories of myopia etiology suggest differences in accommodation and convergence in children who become myopic. Several studies have found differences in accommodation and convergence in myopes and emmetropes. Only one previous study examined accommodation and convergence before youth onset myopia. METHODS: A cohort of initially emmetropic children was given eye and vision examinations at 6-month intervals for 3 years. The data collected included fusional vergence ranges at 4 m and 40 cm, and relative accommodation findings with a target at 40 cm. These clinical accommodation and vergence test findings were compared in children who became myopic (became-myopic group) to children who did not (remained-emmetropic group). The mean test findings were used to construct a zone of clear single binocular vision (ZCSBV) for each group. RESULTS: The midpoint between the 40-cm fusional vergence range blur points was more convergent in the became-myopic group (+5.8 delta compared to +3.2 delta in the remained-emmetropic group; p < 0.004). The positive relative accommodation (PRA) finding was less in magnitude in the became-myopic group (-1.46 D) than in the remained-emmetropic group (-2.04 D), the difference being significant at he 0.02 level. CONCLUSIONS: The ZCSBV near blur point were more convergent in the children who became myopic as indicated by the more convergent position of the midpoint of the near fusional vergence range. The lower value for the PRA test in the became-myopic group duplicates the same result reported in a previous study for retrospective longitudinal private practice data. PMID- 8728495 TI - Clinical findings before the onset of myopia in youth: 3. Heterophoria. AB - BACKGROUND: Some contemporary theories of the etiology of youth onset myopia suggest differences in accommodation and convergence function. Previous studies have related esophoria to various aspects of myopia development. This paper examines whether differences in phorias exist before the onset of myopia. METHODS: A cohort of initially emmetropic children had eye and vision examinations at 6-month intervals for a period of 3 years. Dissociated phorias were measured by the von Graefe method. These prospective data were analyzed by: (1) comparing phorias in children who became myopic to those in children who remained emmetropic; (2) determination of sensitivity and specificity of phorias to distinguish between children who became myopic and those who did not; and (3) determining the change in phorias with time. These same analyses were also performed as additional analyses of some previously published retrospective private practice data. RESULTS: Distance phorias were not significantly different in the became-myopic and remained-emmetropic group by the Mann Whitney U test in either the private practice data or the prospective data. The near phoria was more convergent in the became-myopic group than in the remained-emmetropic group in the private practice data (p < 0.02 by Mann Whitney U test). In the prospective data there were proportionately more children who became myopic outside the near phoria range of 3 delta exo to 1 delta eso (p < 0.05 by chi 2 test). In both data sets the near phoria showed an eso shift with time in the children who became myopic, beginning before and continuing after the onset of myopia. The children in the remained-emmetropic groups did not show eso shifts in near phoria. CONCLUSIONS: A near phoria which is not close to orthophoria appears to be a risk factor for youth onset myopia, but sensitivity and specificity were not as high as those for other variables such as corneal power, axial length to corneal radius ratio, positive relative accommodation (PRA), and midpoint of the near fusional vergence range. The onset of myopia is associated with an eso (convergent) shift in the near phoria. PMID- 8728496 TI - Clinical findings before the onset of myopia in youth: 4. Parental history of myopia. AB - BACKGROUND: We conducted a study to compare variables in children who were initially emmetropic and became myopic to those in children who remained emmetropic. This paper examines parental history of myopia in the two groups of children. METHODS: A cohort of initially emmetropic children was given eye and vision examinations at 6-month intervals for 3 years. Data on parental history of myopia were obtained by questionnaire. RESULTS: A greater proportion of children in the became-myopic group had myopic parents compared to children in the remained-emmetropic group. The sensitivity for onset of myopia was 0.95 and specificity was 0.38 if at least one parent was myopic. A sensitivity of 0.36 and a specificity of 0.87 were found for both parents being myopic. There was a significant association between parental history of myopia and development of myopia in the children in the study (p < 0.01 by chi 2 for categorization of whether neither, one, or both parents were myopic for children in the became myopic and remained-emmetropic groups). CONCLUSION: Parental history of myopia is a risk factor for the development of myopia. PMID- 8728497 TI - Effect of oculomotor and other visual skills on reading performance: a literature review. AB - The diagnosis and management of many oculomotor anomalies is within the domain of optometry. Thus, a thorough understanding of these systems and their relation to reading performance is vital. Efficient reading requires accurate eye movements and continuous integration of the information obtained from each fixation by the brain. A relation between oculomotor efficiency and reading skill has been shown in the literature. Frequently, these visual difficulties can be treated successfully with vision therapy. PMID- 8728499 TI - In vivo and in vitro reactions of toluene diisocyanate isomers with guinea pig hemoglobin. AB - Guinea pig hemoglobin (Hb) adducts of 2,4- and 2,6-toluene diisocyanate (2,4- and 2,6-TDI) were individually prepared, each at a Hb tetramer to diisocyanate ratio of 1:1, and compared with adducts of Hb from animals exposed to 1 ppm 2,4-TDI vapor. Each Hb sample was subjected to C4 HPLC chain/heme separation with UV detection. Survey of the LC fractions using an antiserum prepared to a heterologous TDI-protein indicated a difference in chain specificity and product types formed in vitro by the two isomers. Ionspray mass spectrometry (MS) revealed the chemistry of the adduction products. Carbamoylated products, formed from adduction by one TDI molecule (with one isocyanato group hydrolyzed to an amino group), were detected by MS with 2,4-TDI on each chain and with 2,6-TDI on the beta chain. Additionally, a quasi-molecular ion of a bis carbamoylation adduct was noted by MS in the in vitro 2,4-TDI-Hb adduct in the form of TDI-cross linked alpha and beta chains. Ionspray MS analysis of the Hb isolated from guinea pigs exposed in vivo to 2,4-TDI indicated carbamoylation products with both the alpha and beta chains in which one of the two original isocyanato groups had been hydrolyzed to the amine. We also found evidence of an amine-nitroso adduct on the alpha chain in the in vivo sample. These results indicate that at least one of the isocyanato moieties (or a masked derivative) of 2,4-TDI survived passage through the lung, into the serum, and through the erythrocyte membrane to form adducts with Hb that were stable to dialysis, gel filtration, and reversed phase HPLC separation under acidic conditions. The presence of an apparent amine nitroso adduct indicated in vivo formation of 2,4-diaminotoluene, a recognized animal carcinogen. PMID- 8728498 TI - The in vitro methylation of DNA by a minor groove binding methyl sulfonate ester. AB - The preparation of sequence and groove specific DNA methylating agents based on N methylpyrrolecarboxamide subunits appended with an O-methyl sulfonate ester functionality (MeOSO2(CH2)2-Lex) has previously been described [Zhang, Y., Chen, F.-X., Mehta, P., and Gold, B. (1993) Biochemistry 32,7954-7965]. In contrast to simple methyl sulfonate esters, e.g., methyl methanesulfonate (MMS), which predominantly methylate at 7-guanine, MeOSO2-(CH2)2-Lex affords N3-methyladenine (3-MeAde) as its major adduct. Using competitive ELISA determinations, the methylation at major and minor groove sites in calf thymus DNA by MeOSO2(CH2)2 Lex has been precisely quantitated. The yields of N7-methylguanine (7-MeGua), 3 MeAde, and O6-methyldeoxyguanosine (6-Me-dGuo) are 0.424, 3.195, and 0.0027 mmol of adduct/mol of DNA, respectively, using 10 microM MeOSO2(CH2)2-Lex and 100 microM DNA. This compares to 0.773, 0.072, and 0.0033 mmol of adduct/mol of DNA for 7-MeGua, 3-MeAde, and 6-Me-dGuo, respectively, using MMS. The increase in the yield of 3-MeAde due to the minor groove equilibrium binding properties of MeOSO2(CH2)2-Lex is approximately 40-fold relative to MMS. PMID- 8728500 TI - Comparison of crystal structure and theory for 2-amino-1-methyl-6 phenylimidazo[4,5-b]pyridine. AB - The crystal structure of the food mutagen 2-amino-1-methyl-6-phenylimidazo[4,5-b] pyridine (PhIP) has been determined by single-crystal X-ray crystallography. Crystals grown by evaporation of an aqueous solution form in the monoclinic space group P2(1)/n with two molecules of PhIP per asymmetric unit, along with six water molecules. The phenyl groups of these two PhIP molecules have torsion angles of different magnitude with respect to the plane of the imidazopyridine moiety. To maintain centrosymmetry, the crystal also contains an oppositely torsioned symmetry equivalent of each. The amino groups of both PhIP molecules take part in an extensive hydrogen bond network with the water of crystallization, forming long channels through the crystals parallel to the crystallographic b axis. The diffraction results are compared to theoretical calculations of the optimized geometry for a single PhIP molecule in vacuo as well as with water hydrogen-bonded to the exocyclic amine. In general, the agreement between the X-ray crystal structure of PhIP and its theory-derived counterpart in vacuo is within the combined experimental-theoretical uncertainty. The C-N bond to the exocyclic amine and the neighboring C=N imidazole bond are exceptions. This is attributed to the combined neglect of the crystal environment, waters of hydration, and the lack of coplanarity between the imidazole ring and the amine group in the calculations. To address the effect of waters of hydration, additional calculations were performed to optimize the geometry of a PhIP molecule with two water molecules hydrogen-bonded to the exocyclic amine. The resulting C-N exocyclic amine and C=N imidazole bond lengths were closer to those obtained by X-ray diffraction. The accord between theory and experiment demonstrates the utility of applying theory to (1) accurately predict structures of PhIP metabolites and intermediates that are too labile for study by conventional structural techniques such as X-ray crystallography and (2) assist in studying the mechanisms by which PhIP and its metabolites interact with proteins and DNA. PMID- 8728501 TI - Mechanism of acetaminophen-induced hepatotoxicity: covalent binding versus oxidative stress. AB - The hepatotoxicity of acetaminophen is believed to be mediated by the reactive metabolite N-acetyl-p-benzoquinone imine; however, the mechanism by which this metabolite produces the toxicity is unknown. The metabolite, which is both an electrophile and an oxidizing agent, may covalently bind to critical proteins, or it may initiate oxidative damage. We have previously developed a Western blot assay for detection of acetaminophen covalently bound to protein and have reported the relationship between covalent binding and the development of hepatotoxicity. Recently, we developed a Western blot assay for protein aldehyde formation, which may occur via the reactive oxygen species, the hydroxyl radical. In this paper, we have compared covalent binding to protein aldehyde formation. Toxic doses of acetaminophen (400 mg/kg) were administered to mice, and the mice were subsequently killed at 0, 1, 2, 4, and 6 h. Since the oxidizing agent FeSO4 has been reported to potentiate lipid peroxidation when administered with acetaminophen, other mice received FeSO4 (100 mg/kg) plus acetaminophen. Compared to saline-treated control mice, acetaminophen treatment significantly increased serum alanine aminotransferase levels, an index of hepatotoxicity, at 4 and 6 h, but not at 1 or 2 h. Acetaminophen plus FeSO4 treatment of mice significantly increased serum alanine aminotransferase levels at 2, 4, and 6 h compared to controls. Levels of alanine aminotransferase in serum of acetaminophen plus ferrous sulfate-treated mice were higher at 4 and 6 h than those of acetaminophen treated mice, but not significantly different. FeSO4 alone did not increase alanine aminotransferase levels. Western blot assays revealed that acetaminophen did not cause an increase in protein aldehydes over control at any time, nor did acetaminophen plus FeSO4; however, FeSO4 alone increased the intensity of staining of the immunoblot for protein aldehydes over control at all times after 0 time. Acetaminophen-protein adducts were detected in acetaminophen- and acetaminophen plus FeSO4-treated mice. In vitro experiments indicated that FeSO4 plus tert-butyl hydroperoxide in the presence of bovine serum albumin increased protein aldehyde formation. Inclusion of acetaminophen in the incubation mixture inhibited protein oxidation of bovine serum albumin in a concentration dependent manner. The data indicate that acetaminophen quenches protein oxidation, presumably by reacting with the hydroxyl radical. These data are consistent with the theory that acetaminophen covalent binding is the primary mechanism of toxicity and argue against a role for protein oxidation in acetaminophen hepatotoxicity. PMID- 8728502 TI - Benzo[c]phenanthrene-DNA adducts in mouse epidermis in relation to the tumorigenicities of four configurationally isomeric 3,4-dihydrodiol 1,2-epoxides. AB - P-Postlabeling assays were used to monitor the binding to epidermal DNA that resulted from the application of each of the four configurational isomers of benzo[c]phenanthrene 3,4-dihydrodiol 1,2-epoxide to mouse skin in vivo. For three of these configurational isomers, there was a reasonable correlation between the relative level of binding to epidermal DNA and the known tumorigenic effects of these compounds. However, for the 4(S),3(R)-dihydrodiol 2(S),1(R)-epoxide, the tumorigenic response was considerably greater in relation to the level of DNA modification than was the case for the other isomers. This greater tumorigenic response was consistent with previous observations indicating that this isomer was more mutagenic, at equivalent levels of DNA modification, than the other two tumorigenic dihydrodiol epoxides. Additionally, the 4(S),3(R)-dihydrodiol 2(S),1(R)-epoxide reacts with DNA to generate predominantly (approximately 80%) adducts on the amino group of adenine residues. These findings might imply a greater intrinsic biological effect of such adenine adducts with respect to the other major adduct formed on the amino group of guanine residues. PMID- 8728503 TI - Synthesis of N2-(4-hydroxyphenyl)-2'-deoxyguanosine 3'-phosphate: comparison by 32P-postlabeling with the DNA adduct formed in HL-60 cells treated with hydroquinone. AB - A new adduct has been isolated from the reaction of guanosine 3'-phosphate and p benzoquinone. The structure of this adduct has been determined as N2-(4 hydroxyphenyl)-guanosine 3'-phosphate. 32P-Postlabeling showed that this adduct is similar to the DNA adduct formed in HL-60 cells treated with hydroquinone. For comparison with the corresponding deoxyribonucleotide, a synthetic procedure was developed for the preparation of N2-substituted derivatives of 2'-deoxyguanosine 3'-phosphate. 2-Bromo-2'-deoxyinosine 3'-phosphate was synthesized with a combination of synthetic and enzymatic methods. Reaction of 2-bromo-2' deoxyinosine 3'-phosphate with 4-hydroxyaniline gave N2-(4-hydroxyphenyl)-2' deoxyguanosine 3'-phosphate. Using 32P-postlabeling, we compared this product with the DNA adduct produced in HL-60 cells treated with hydroquinone. The results of these studies suggest that the DNA adduct formed in HL-60 cells treated with hydroquinone corresponds to N2-(4-hydroxyphenyl)-2'-deoxyguanosine 3'-phosphate. PMID- 8728504 TI - Inhibition of purified and membrane-bound flavin-containing monooxygenase 1 by (N,N-dimethylamino)stilbene carboxylates. AB - (E)-[2-(4-(Dimethylamino)phenyl)vinyl]benzenes bearing a nitrile or carboxyl group in the 2', 3', or 4' position were synthesized and tested for substrate activity with purified pig liver flavin-containing monooxygenase (FMO1). Although the nitrile derivatives were too insoluble to saturate the catalytic site at pH 7.4, they appeared to be substrates with K(m)'s somewhat above their maximum solubility (approximately equal to 0.1 mM) in the assay medium. Of the three carboxylic acid analogs, (E)-4-[2-(4(dimethylamino)phenyl)vinyl]benzoic acid had no detectable water solubility at pH 7.4, and measurements were restricted to (E) 3-[2-(4-(dimethylamino)phenyl)vinyl]benzoic acid (DS3CO) and (E)-2-[2-(4 (dimethylamino)phenyl)vinyl]benzoic acid (DS2CO). While DS3CO and DS2CO were substrates, they also inhibited FMO1 turnover. DS3CO was the more effective inhibitor, and at 2 mM it inhibited FMO1 and microsomal-catalyzed oxidation of methimazole (N-methyl-2-mercaptoimidazole) by 80-90%. Kinetic studies indicated that the aminostilbene carboxylates were noncompetitive with both the xenobiotic substrate, methimazole, and NADPH. However, inhibition constants calculated from double reciprocal plots of velocity vs NADPH were K(i)(comp) 130 and 150 microM for DS3CO and DS2CO, respectively, whereas the uncompetitive Ki's were 10-15 times higher, which suggests that inhibition of NADPH binding may be primarily responsible for inhibition of FMO1 by the aminostilbene carboxylates. This model is also consistent with inhibition of cyclohexanone monooxygenase, a bacterial analog of FMO. DS3CO and DS2CO were again noncompetitive with methimazole but primarily competitive with NADPH. The aminostilbene carboxylates had no detectable effects on activity of pig or rat liver NADPH-cytochrome P450 reductase, which suggests that they are not nonspecific flavoprotein antagonists. PMID- 8728505 TI - Photochemical formation of singlet molecular oxygen in illuminated aqueous solutions of several commercially available sunscreen active ingredients. AB - Evidence is presented for the photochemical formation of singlet molecular oxygen (1O2) in air-saturated aqueous solutions of several sunscreen active ingredients using sunlight-range illumination. This is of significance because (1) 1O2 is known to be cytotoxic, and (2) there have been several reports of toxic effects associated with the use of some sunscreens; most notably, with p-aminobenzoic acid (PABA). Illuminated aqueous solutions of PABA, 2-ethylhexyl p (dimethylamino)benzate (ODPABA), 2-hydroxy-4-methoxybenzophenone (BZ3), 2,2' dihydroxy-4-methoxybenzophenone (BZ8), 2-ethylhexyl 2-cyano-3,3-diphenylacrylate (OCR), 2-ethylhexyl p-methoxycinnamate (OMC), and 2-ethylhexyl salicylate (OCS) were evaluated individually for 1O2 formation. Furfuryl alcohol (FFA), a well known chemical trap for 1O2, was added to each of the aqueous sunscreen solutions. The FFA was consumed when solutions of PABA, ODPABA, OMC, and OCR were illuminated, but no loss of FFA other than by direct photolysis occurred in solutions of BZ3, BZ8, or OCS. There was also no significant loss of FFA in any of these solutions kept in the dark. Further evidence for the formation of 1O2 in illuminated aqueous sunscreen solutions is provided by the results of experiments in which individual solutions containing sunscreen active ingredients and FFA that were diluted with D2O exhibited an increased rate of FFA consumption while the addition of azide ion (N3-) reduced the rate of FFA consumption. Continuous sunlight-range illumination of aqueous PABA solutions produced significantly higher steady-state concentrations of 1O2 than in solutions containing any of the other sunscreen active ingredients evaluated. The substituted benzophenone compounds (BZ3 and BZ8) and the salicylate-based compound (OCS) not only appear to produce no 1O2, but they also appear to produce no other reactive oxidant species that are capable of consuming FFA. This indicates that BZ3, BZ8, and OCS may be peferable, from the standpoint of toxic oxidant formation, for use as sunscreen active ingredients when compared to the other compounds evaluated in this study. PMID- 8728506 TI - Toxicity of organic chemicals to fathead minnow: a united quantitative structure activity relationship model and its application. AB - A general and practical relation has been established between structure and activity of organic chemicals based on target theory. This relation was tested by examining the correlations between the acute toxicity and the physicochemical properties for fathead minnow, which showed that this relation can explain the observed toxicity data and phenomena successfully. The equations are then used to estimated the acute toxicity data of organic chemicals, and the predicted values are close to observed values. PMID- 8728507 TI - Identification of the heme adduct and an active site peptide modified during mechanism-based inactivation of rat liver cytochrome P450 2B1 by secobarbital. AB - The olefinic barbiturate secobarbital (SB) is a sedative hypnotic known to be a relatively selective mechanism-based inactivator of rat liver cytochrome P450 2B1. Previous studies have demonstrated that such inactivation results in prosthetic heme destruction and irreversible drug-induced protein modification, events most likely triggered by P450 2B1-dependent oxidative activation of the olefinic pi-bond. However, the precise structure of the SB-modified heme and/or the protein site targeted for attack remained to be elucidated. We have now isolated the SB-heme adduct from P450 2B1 inactivated by [14C]SB in a functionally reconstituted system and structurally characterized it by electronic absorption spectroscopy and tandem collision-induced dissociation (CID), matrix assisted laser desorption ionization on time of flight (MALDI-TOF), and liquid secondary ion mass spectrometry in the positive mode (+ LSIMS) as the N-(5-(2 hydroxypropyl)-5-(1-methylbutyl)barbituric acid)protoporphyrin IX adduct. The [14C]SB-modified 2B1 protein has also been isolated from similar inactivation systems and subjected to lysyl endopeptidase C (Lys-C) digestion and HPLC-peptide mapping. A [14C]SB-modified 2B1 peptide was thus isolated, purified, electrotransferred onto a poly-(vinylidene) membrane, and identified by micro Edman degradation of its first N-terminal 17 residues (S277NH(H)TEFH(H)ENLMISLL293) as the Lys-C peptide domain comprised of amino acids 277-323. This peptide thus includes the peptide domain corresponding to the distal helix I of P450 101, a region highly conserved through evolution, and which is known not only to flank the heme moiety but also to intimately contact the substrates. This finding thus suggests that SB-induced protein modification of P450 2B1 also occurs at the active site and, together with heme N-alkylation, contributes to the SB-induced mechanism-based inactivation of P450 2B1. PMID- 8728508 TI - Metabolic activation of PCBs to quinones: reactivity toward nitrogen and sulfur nucleophiles and influence of superoxide dismutase. AB - Polychlorinated biphenyls (PCBs) may undergo cytochrome P-450-catalyzed hydroxylations to form chlorinated dihydroxybiphenyl metabolites. When the hydroxyl groups are ortho or para to each other, oxidation to a quinone may be catalyzed by peroxidases present within the cell. In order to study the reactivity of PCB-derived quinones, selected chlorophenyl 1,2- and 1,4 benzoquinones were synthesized and characterized, including their reduction potentials against a saturated calomel electrode. Two quinones, 4-(4' chlorophenyl)-1,2-, and 4-(3',4'-dichlorophenyl)-1,2-benzoquinone, were obtained via the oxidation of the corresponding dihydroxybiphenyls with 2,3-dichloro-5,6 dicyano-1,4-benzoquinone. Six 1,4-benzoquinones were synthesized via the Meerwein arylation: 2-(2'-chlorophenyl)-1,4-, 2-(3'-chlorophenyl)-1,4-, 2-(4' chlorophenyl)-1,4-, 2-(2',5'-dichlorophenyl)-1,4-, 2-(3',4'-dichlorophenyl)-1,4-, and 2-(3',5'-dichlorophenyl)-1,4-benzoquinone. As a model study, the rate of reactivity of 2-(4'-chlorophenyl)-1,4-benzoquinone toward the nitrogen nucleophiles glycine, L-arginine, L-histidine- and L-lysine was determined under pseudo-first-order conditions at pH 7.4. The rate constants ranged from 0.45 to 0.75 min-1 M-1. Higher rates were obtained under conditions of higher pH. Two reaction products were identified as the 5- and 6-ring addition products in the ratio of 1:4. In contrast, the reaction of 2-(4'-chlorophenyl)-1,4-benzoquinone with the sulfur nucleophiles glutathione or N-acetyl-L-cysteine was instantaneous. The major product of the reaction of glutathione with 2-(4' chlorophenyl)-1,4-benzoquinone was also the 6-ring addition product. The hydroquinone thioether could be enzymatically reoxidized to the quinone thioether. Also, the influence of atmospheric oxygen and superoxide dismutase on the rates of the following horseradish peroxidase/H2O2-catalyzed oxidations was investigated: 3,4-dichloro-2',5'-dihydroxybiphenyl to 2-(3',4'-dichlorophenyl) 1,4-benzoquinone and 3,4-dichloro-3',4'-dihydroxybiphenyl to 4-(3',4' dichlorophenyl)-1,2-benzoquinone. While the presence or absence of atmospheric oxygen did not alter the rates of the oxidation reactions, the presence of superoxide dismutase significantly increased the rates of both oxidation reactions, having the greater effect on the oxidation of the 1,4-hydroquinone. These data show that PCB-derived quinones react with both nitrogen and sulfur nucleophiles of the cell and may explain, in part, the toxic effects of individual PCBs and PCB formulations, such as glutathione depletion, oxidative stress, and cell death. PMID- 8728509 TI - Improved strategies for postoligomerization synthesis of oligodeoxynucleotides bearing structurally defined adducts at the N2 position of deoxyguanosine. AB - Improved methodology has been developed for preparation of oligodeoxynucleotides bearing adducts on the N2 position of guanine in which the adduction reaction is carried out in homogeneous solution rather than while the oligonucleotide is immobilized on a solid matrix. The methodology utilizes a new synthon, 2-fluoro O6-(trimethylsilylethyl)-2'-deoxyinosine (3). Nucleoside 3 is stable to the conditions of oligonucleotide synthesis, but the O6 protection is eliminated under very mild conditions following displacement of the 2-fluoro group by amine nucleophiles. Oligonucleotides containing 3 could be removed from the solid support by treatment with 0.1 M NaOH (8 h, rt) without disruption of 3. Reaction of the crude, partially deprotected oligonucleotide with (R)-2-amino-2 phenylethanol in homogeneous solution, followed by removal of the remaining protective groups with NH4OH (60 degrees C, 8 h) and then 0.1% acetic acid, gave the adducted oligonucleotide in good purity and yield. Alternatively, fully deprotected oligonucleotide containing 3 could be prepared by use of labile phenoxyacetyl-type protecting groups on the exocyclic amino groups. PMID- 8728510 TI - Regioselectivity and quantitative structure-activity relationships for the conjugation of a series of fluoronitrobenzenes by purified glutathione S transferase enzymes from rat and man. AB - Quantitative structure-activity relationships (QSAR's) are described for the rate of conjugation of a series of fluoronitrobenzenes with cytosolic as well as with two major alpha and mu class enzymes of rat and human liver, viz., glutathione S transferases (GST) 1-1, 3-3, A1-1, and M1a-1a. For all purified enzymes studied, the natural logarithm of the rate of conversion of the fluoronitrobenzenes correlates with both the calculated reactivity of the fluoronitrobenzenes for an electrophilic attack (i.e., E(LUMO)) and the calculated relative heat of formation for formation of the respective Meisenheimer complex intermediate (delta delta HF). In addition, the regioselectivity of the reaction was determined and compared. The results obtained strongly support the conclusion that chemical reactivity of the fluoronitrobenzenes is the main factor determining the outcomes of their conversion by all glutathione S-transferase enzymes. The regioselectivities vary only a few percent from one enzyme to another, whereas QSAR lines for all purified enzymes are in the same region and run parallel. This indicates that in the overall reaction the nucleophilic attack of the thiolate anion on the fluoronitrobenzenes, leading to formation of the Meisenheimer complex, is the rate-limiting step in the overall catalysis. The fact that chemical reactivity of the fluoronitrobenzenes is the main factor in setting the outcomes of the overall conversion by the different glutathione S transferase enzymes implies that extrapolation from rat to results of other species including man, and also from one individual to another, must be feasible. That this is actually the case is clearly demonstrated by the results of the present study. PMID- 8728511 TI - Characterization of chlorinated adducts of hemoglobin and albumin following administration of pentachlorophenol to rats. AB - Five cysteinyl adducts (including one with multiple isomeric forms) of hemoglobin (Hb) and albumin (Alb) have been characterized in the blood of Sprague-Dawley rats following administration of pentachlorophenol (PCP). Three of these adducts were formed by multiple substitution reactions of tetrachloro-1,4-benzoquinone (Cl4-1,4-BQ) and its products, and two arose from reactions of tetrachloro-1,4 benzosemiquinone (Cl4-1,4-SQ) and tetrachloro-1,2-benzosemiquinone (Cl4-1,2-SQ). Adducts of tetrachloro-1,2-benzoquinone (Cl4-1,2-BQ) were not observed. Regarding adducts of Cl4-1,4-BQ and its products, specific structures were assigned to monosubstituted, disubstituted, and trisubstituted adducts of Hb and Alb following modification of rat blood with Cl4-1,4-BQ (0-45 microM) in vitro and after metabolism of PCP (0-40 mg/kg body weight) in Sprague-Dawley rats, in vivo. The formation of all adducts was linear over the ranges tested, with Alb adducts being more abundant than Hb adducts. The levels of the adducts measured were in the following order: monosubstituted > disubstituted > trisubstituted. The observation that Cl4-1,4-BQ can produce multisubstituted adducts with proteins suggests that protein-protein cross links may be formed, with inherent toxicological implications. Regarding adducts of the semiquinones (detected only in vivo), linear production of Hb and Alb adducts was observed with increasing dosage of PCP for adducts of both Cl4-1,4-SQ and Cl4-1,2-SQ. Higher levels of the semiquinone adducts were observed in Hb than in Alb, in contrast to the results with the quinone adducts. In a separate in vivo experiment (20 mg PCP/kg body weight), where animals were sacrificed at intervals up to 336 h postadministration, adducts were eliminated at rates which were comparable among the different adducts of a given protein. PMID- 8728512 TI - Effects of pH, temperature, and chemical structure on the stability of S-(purin-6 yl)-L-cysteine: evidence for a novel molecular rearrangement mechanism to yield N (purin-6-yl)-L-cysteine. AB - The stability of S-(purin-6-yl)-L-cysteine (SPC), a kidney-selective prodrug of 6 mercaptopurine and a putative metabolite of 6-chloropurine, was investigated under various pH and temperature conditions. At room temperature, the half-life (t 1/2) of SPC at either highly acidic (pH 3.6) or basic conditions (pH 9.6) was longer than at neutral or slightly acidic or basic conditions (pH 5.7-8.75). The primary degradation product, N-(purin-6-yl)-L-cysteine (NPC), was isolated using Sephadex LH-20 chromatography and characterized by 1H NMR and FAB/MS after derivatization with 2-iodoacetic acid. These results reveal novel stability requirements and implicate the cysteinyl amino group and the purinyl N-1 nitrogen in the mechanism of SPC rearrangement to NPC. Further evidence for this hypothesis was provided by the findings that the stability of SPC in phosphate buffer (pH 7.4) at 37 degrees C was similar to that of S-(guanin-6-yl)-L cysteine, whereas S-(purin-6-yl)-N-acetyl-L-cysteine and S-(purin-6 yl)glutathione which have their cysteine amino groups blocked were much more stable than SPC. S-(Purin-6-yl)-L-homocysteine (SPHC) was also more stable than SPC, possibly because the formation of a 6-membered ring transition state as would be expected with SPHC is kinetically less favored than the formation of a 5 membered ring transition state as would be expected with SPC. These results may explain previous in vivo metabolism results of SPC and its analogs and may contribute to a better understanding of stability of structurally related cysteine S-conjugates. PMID- 8728513 TI - Chemical and immunochemical comparison of protein adduct formation of four carboxylate drugs in rat liver and plasma. AB - Carboxylate drugs usually form acyl glucuronide conjugates as major metabolites. These electrophilic metabolites are reactive, capable of undergoing hydrolysis, rearrangement, and covalent binding reactions to proteins. The last-mentioned property has the potential to initiate immune and other toxic responses in vivo. In this study, we compared the extent and pattern of covalent adduct formation in plasma and livers of rats dosed with the nonsteroidal anti-inflammatory drugs (NSAIDs) zomepirac (ZP) and diflunisal (DF), the hypolipidemic agent clofibric acid (CA), and the anti-epileptic agent valproic acid (VPA). These drugs form acyl glucuronides with diverse intrinsic reactivities (apparent first order degradation t 1/2 values of 0.5, 0.6, 3, and 60 h, respectively). Rats were dosed iv twice daily for 2 days (50 mg/kg for ZP, DF, and CA, 150 mg/kg for VPA). Chemical analysis of tissues obtained 6 h after the last dose revealed adduct concentrations of 0.31, 0.44, 0.28, and 0.05 micrograms of drug equivalents/mL of plasma and 2.21, 2.31, 0.96, and 0.96 micrograms of drug equivalents/g of liver for ZP, DF, CA and VPA treatments, respectively. For both plasma and liver, the higher concentrations of adducts were found with ZP and DF, which have the more reactive glucuronides. The low concentrations of VPA adducts found in plasma were in keeping with the very low reactivity of its glucuronide. In liver, however, VPA adducts achieved concentrations of the same order of magnitude as the other drugs and were accompanied by adducts of the (E)-2-en metabolite of VPA at 0.38 micrograms of VPA equivalents/g of liver. The liver data for VPA can be explained by an acyl CoA/beta-oxidation pathway of adduct formation in addition to that from acyl glucuronidation. Immunoblotting using rabbit polyclonal antisera raised against synthetic drug-protein adducts revealed major bands at 110, 140, and approximately 200 kDa in livers of ZP- and DF-treated rats. A fourth major band at 70 kDa in ZP-treated liver had the same apparent molecular weight as the only major band detected in CA-treated liver. A 140 kDa band was detected in liver tissue from VPA-treated rats, as well as several lower molecular weight bands. In plasma, the antisera specifically detected drug-modified serum albumin in samples from rats treated with ZP, DF, and CA, but not VPA. The results with this small series of carboxylate drugs suggested that (a) adduct concentrations in plasma but not liver could be related to acyl glucuronide reactivity, (b) while some modified proteins detected were common, the pattern of modification varied from drug to drug, and (c) caution should be exercised in attributing adduct formation exclusively to the acyl glucuronidation pathway. PMID- 8728515 TI - Academic research debated. PMID- 8728514 TI - Conformational aspects of glutathione conjugates of chlorinated alkenes: a computational study. AB - The nephrotoxicity of halogenated alkenes is due to the beta-lyase mediated bioactivation of the hepatic glutathione (GS) conjugate to mutagenic or cytotoxic reactive species in kidney. Experimental evidence obtained for regioisomers and geometric isomers of haloalkene GS conjugates indicates that different isomers may be metabolized and excreted at different rates, follow different metabolic pathways, and exhibit different toxicities. Computational methods were applied in the present work to a conformational study of GS-haloalkene conjugates to determine the relative stabilities of possible regioisomers and geometric isomers of the conjugates. The halogenated alkenes studied were 1,1,2-trichloroethylene (TCE), hexachloro-1,3-butadiene (HCBD), and 1,1,2-trichloro-3,3,3-trifluoro-1 propene (TCTFP). Calculated energies of GS conjugate products were used to approximately infer relative product abundance under synthetic and in vivo conditions. This approach neglects differential solvent effects and enzyme selectivity and assumes a late transition state for GS conjugation and/or some thermodynamic control of the conjugation process. Relative population predictions of GS conjugate isomers, based on computed energies, were in agreement with experimental synthetic and in vivo isomer determinations in the case of TCE, where careful analytical characterization of the isomers was definitive. In the case of HCBD, where analytical determinations were not performed and isomer assignments were based on general reactivity concepts, calculations from the present study supported one GS conjugate isomer assignment and disagreed with the other. Finally, in the case of TCTFP, the calculations predicted that three isomers would have similar populations, whereas only two were detected in the experimental study. PMID- 8728516 TI - Teaching professional boundaries in the physician-patient relationship. PMID- 8728517 TI - Abortion training in family practice residency programs. AB - BACKGROUND: Despite a relatively fixed demand for services, access to abortion services has become increasingly limited, in part due to fewer training opportunities for OB-GYN residents. The recommended core educational guidelines for family practice residents include voluntary interruption of pregnancy up to 10 weeks gestation, but the availability of and interest in such training have not been reported. METHODS: A survey questionnaire assessing availability of abortion training, percentage of residents participating, program demographics, and resident interest in training was sent to all US family practice residency program directors and randomly selected third-year residents during the 1993-1994 academic year: A total of 301 of 399 (75%) program directors and 253 of 399 (63%) third-year residents completed the questionnaire. RESULTS: Twelve percent of programs nationwide offered abortion training during 1993-1994. Western programs were more likely to offer training (18 of 50 or 36%) than Northeastern (7 of 55 or 12%), Midwestern (6 of 98 or 6%), or Southern programs (4 of 98 or 4%). When available, 45% of trainees chose to participate. Training was primarily in suction curettage up to 12 weeks gestational age. The median duration of training was 4 weeks. Training was provided in both freestanding clinics and hospitals. Religious hospitals were less likely to offer training. CONCLUSIONS: About one of eight family practice residency programs nationwide offer training in abortion. When offered, nearly half of family practice residents choose to participate in this training. Interested family practice residents could be trained to fill an increasing need for physicians willing and able to perform first-trimester abortions. PMID- 8728518 TI - Family medicine in a culturally diverse world: a solution-oriented approach to common cross-cultural problems in medical encounters. AB - BACKGROUND: Using cultural sensitivity in the training of family practice residents generally results in positive consequences for patient care. However, certain potential problems associated with cross-cultural educational efforts deserve examination, including patient stereotyping, assumptive bias, and the confounding of ethnicity with class and socioeconomic status. Even awareness of these pitfalls may not guarantee physician avoidance of other barriers to effective patient care, such as communication difficulties, diagnostic inaccuracies, and unintentional patient exploitation. Despite these complications, future family physicians must continue to participate in educational activities that increase sensitivity toward and understanding of patients of different ethnicities. This article discusses certain features characteristic of the ways in which cultural variables operate in the doctor patient encounter and identifies specific ways in which residents can successfully elicit and use cultural knowledge to enhance patient care. PMID- 8728519 TI - A curriculum for teaching family practice residents how to manage skin lesions. AB - BACKGROUND AND OBJECTIVES: A curriculum was developed to teach family practice residents how to manage skin lesions as part of an outpatient rotation during which family practice residents are taught procedures by family medicine faculty. The curriculum can be used as a template for teaching and assessing competency for any family practice outpatient procedure. METHODS: The curriculum was pilot tested by eight second-year family practice residents and evaluated by experts in curriculum design and the methods for teaching outpatient procedural skills. Procedure checklists were used to evaluate competency. RESULTS: The residents scored higher on their posttests than on their pretests. The residents, implementing faculty, and outside reviewers rated the curriculum design highly. Performance of procedures did not require longer times when accomplished through this educational experience. CONCLUSIONS: This curriculum can be used to teach family practice residents how to manage skin lesions. Residents were deemed competent to perform a procedure if they successfully followed all of the steps on the procedure checklist. A specific minimum number of procedures was not required to establish competency. PMID- 8728520 TI - Time use and physicians' exploration of the reason for the office visit. AB - BACKGROUND: Patient-centered clinical methods encourage physicians to explore the patient's reasons for the visit. Practitioners worry that this behavior may be inefficient. This study piloted a method to examine how the number of problems managed and time per problem vary with time spent exploring a patient's views. METHODS: Exploration of a patient's views, medical investigation, explanation, and management were mapped against time for a defined agenda from videotapes of eight second-year residents seeing three simulated patients. Performance of three of four tasks defined completion of work on an agendum. Visit length, time per agendum completed, and proportion of agenda completed were compared with time per task using regression methods. RESULTS: Time on exploration correlated with visit length and proportion of agenda completed, but other tasks did not. Time per agendum completed was unrelated to time on exploration. CONCLUSIONS: Visits were longer when more time was spent exploring the patient's reason for the visit because the physician tackled more problems. Time required per problem was not significantly increased. Task/agenda mapping is worth further development to examine differences in comprehensiveness and efficiency among practice styles. PMID- 8728521 TI - Using explanatory models to understand chronic symptoms of Cambodian refugees. AB - BACKGROUND: Case reports indicate that understanding patients' "explanatory models" can improve diagnosis, patient compliance, and satisfaction. However, few studies have examined explanatory models within specific cultural groups to understand their symptoms. METHODS: Seventy-six older Cambodian refugees were interviewed using measures of symptom frequency, explanatory models, help-seeking patterns, and use of traditional Cambodian remedies. Psychiatric patients were compared with non-patients. RESULTS: Headache was the most common symptom, reported by 58% of those interviewed, and also the most frequent chief complaint (41%). Headache was reported in both groups but was reported by significantly more psychiatric patients. Headache was positively associated with the diagnosis of depression. More than half explained their headaches as a result of sadness, grief, and anxiety. Explanatory models did not differ significantly between groups. All participants maintained Cambodian medical beliefs, and 83% used traditional Cambodian remedies. CONCLUSIONS: Results suggest a high prevalence of headache among older Cambodian refugees, for whom it may be a manifestation of depression. Emphasis on this somatic complaint does not preclude patients' recognition of psychological factors. Addressing explanatory models can help clinicians determine underlying etiology and understand challenging patients. PMID- 8728522 TI - Practical issues in conducting small-area variation analysis. AB - BACKGROUND AND OBJECTIVES: This paper provides a detailed examination of the conduct of a small-area variation analysis (SAVA) health services research project to aid other researchers in performing and interpreting such projects. STUDY CONDUCT: SAVA was performed on hospital admissions with suspected acute cardiac ischemia in the state of Michigan, and the details of each of the steps are presented. The study team consisted of the principal investigator, a biostatistician, a health economist, a medical geographer, and a systems analyst. Data were obtained from the state hospital association and from a commercial demographics source. Small areas for analysis were defined by a patient origin clustering method. Crude area rates were adjusted for age and gender composition of small areas. Multivariate Poisson regression that included sociodemographic variables was performed on adjusted rates. Issues in interpretation of SAVA and practical barriers to primary care SAVA are discussed. PMID- 8728523 TI - Family practice training in Estonia. AB - BACKGROUND AND OBJECTIVES: The collapse of the Communist regime has required many newly independent countries to reform their health care systems. This article presents the unique experience of Estonia's development of family practice training and the introduction of this specialty to the existing health care system. Family practice training programs began in 1991 and include several pathways to certification in family practice. For physicians currently working as district doctors, pediatricians, gynecologists, or emergency doctors, there is the possibility of part-time retraining while continuing their work as primary care physicians. For new graduates of the medical school, there is an internship and residency in family practice or the option of part-time or in-service vocational training while maintaining a clinical practice. An important aim of the programs is to develop the identity of family physicians. The success and the failures of the Estonian experience in launching this project may be useful to other countries facing health care reform. PMID- 8728524 TI - Abortion training in family medicine. PMID- 8728525 TI - Fire in the belly: doing what it takes to produce excellent research. PMID- 8728526 TI - STFM Core Curriculum Guidelines. Recommended core curriculum guidelines on culturally sensitive and competent health care. PMID- 8728527 TI - Electromyographic and kinematic analysis of graded treadmill walking and the implications for knee rehabilitation. AB - Muscle activity, joints, angles, and heart rate during uphill walking were compared for application in knee rehabilitation. The objectives of this study were to quantify muscle activation levels at different treadmill grades and to determine the grade(s) at which knee range of motion would not further compromise the joint. Average and peak electromyographic activity of the quadriceps (vastus medialis oblique and vastus lateralis) and hamstrings (biceps femoris and medial hamstrings (semimembranosus/semitendinosus)] was recorded during walking at 0, 12, and 24% grade. Six subjects (age = 28.5 +/- 3.7 years, stature = 1.79 +/- .05 m, and mass = 74.7 +/- 7.9 kg) walked at self-selected speeds at each grade while ankle, knee and hip angles, heart rate, and electromyographic activity (surface electrodes) were recorded. Maximum voluntary contractions provided a relative reference for the electromyographic activity during walking. Average and peak electromyographic activity increased significantly across grades for the vastus medialis oblique (125 and 154%), vastus lateralis (109 and 139%), and biceps femoris (53 and 46%), but remained similar for the medial hamstrings. Maximum knee flexion at heel strike increased significantly with grade. Despite decreased self-selected speeds with increasing grade, there were significant increases in heart rate across grades. The results of this study provide a basic understanding of the quadriceps and hamstrings activity levels, lower extremity joint range of motion, and cardiovascular requirements of graded treadmill walking in normal subjects. The results also suggest that a grade just greater than 12% may be most beneficial for knee rehabilitation to minimize patellofemoral discomfort or potential strain on the anterior cruciate ligament. The benefits achieved through this functional activity encourage its implementation in rehabilitation and provide a basis for comparison with injured patients. PMID- 8728528 TI - The effect of extraneous movements on peak torque and constant joint angle torque velocity curves. AB - During isokinetic knee testing, some subjects flex the trunk and contralateral knee during knee extension and the trunk and contralateral knee during knee flexion. This investigation assessed the effect of these extraneous movements on peak and constant joint angle torque (15 degrees below horizontal). Twenty-one subjects performed isokinetic knee extensions and flexions at 60, 180, and 300 degrees/sec under stabilized and nonstabilized conditions. The nonstabilized testing was performed with intentional trunk and contralateral leg movements. The stabilized condition resulted in greater torque values than the nonstabilized condition for extension and flexion for both peak torque and constant joint angle torque. The angle at which flexion peak torque occurred was further from full extension at 60 and 180 degrees/sec under the nonstabilized condition, while no differences were found for extension. These results indicate that extraneous movements can decrease torque output, which may be due to changes in muscle length caused by the extraneous movements. PMID- 8728529 TI - Interinnominate motion and symmetry: comparison between gymnasts and nongymnasts. AB - Some clinical approaches to the treatment of low back pain evaluate and treat observed asymmetries of pelvic posture and motion. Scientific evidence suggests the motion available between the innominate bones is small and variable in nature. The purposes of this investigation were 1) to determine if interinnominate motion of subjects without low back pain was symmetrical in reciprocal test posture combinations, 2) to assess innominate bone symmetry in standing, and 3) to determine if a difference in the magnitude of interinnominate motion was present between a subject group which performs more frequent flexibility activities compared with a subject group representing the general population. Thirty-four subjects (eight male gymnasts, nine female gymnasts, eight male nongymnasts, and nine female nongymnasts) were evaluated in standing and three other reciprocal postures (modified standing, modified sitting, and half-kneeling). In each posture, the Metrecom Skeletal Analysis System was used to obtain coordinates for the anterior and posterior iliac spines. Projection angles were used to determine the relative positions of the right and left innominate bones. Results suggest that stand to right modified standing and stand to left modified standing oblique sagittal interinnominate composite motions were symmetrical, stand to right modified sitting and stand to left modified sitting oblique sagittal interinnominate composite motions were asymmetrical, and stand to right half-kneel and stand to left half-kneel oblique sagittal interinnominate composite motions' symmetrical properties were mixed depending on the group. Gymnasts as a group were found to have asymmetrically positioned innominate bones while nongymnasts as a group had symmetrically positioned innominate bones. PMID- 8728530 TI - The effects of semirigid Air-Stirrup bracing vs. adhesive ankle taping on motor performance. AB - The impact on athletic performance is a critical consideration when choosing ankle support devices for competitive athletes. The purpose of this study was to determine if Air-Stirrup ankle bracing and adhesive ankle taping differed in their effects on motor performance. An additional purpose was to determine if there was a subjective preference toward the use of one support method over the other. Twenty-six male athletes performed an agility run, a 40-yard sprint, and a vertical jump while wearing: 1) adhesive tape, 2) Air-Stirrup brace, and 3) no support (control). Observed data suggest that both taping and bracing have no substantial effect on agility, sprinting speed, or vertical jumping ability. The difference in perceived comfort ratings reported by subjects with respect to support system shows that subjects were more comfortable using the brace over a standard ankle taping procedure. These results suggest that both the Air-Stirrup brace and conventional tape do not impede athletic performance. This study may assist athletes and clinicians in making a more informed selection of a prophylactic ankle support device when considering the effect on performance and comfort. PMID- 8728531 TI - Effect of orthotics on postural sway following inversion ankle sprain. AB - Orthotic devices have been shown to successfully modify selected aspects of lower extremity mechanics and enhance foot stability during the support phase of running. It was hypothesized that orthotic intervention would relieve excessive strain on the ankle ligaments and reduce postural sway, especially in subjects with acute ankle sprains. The primary purpose of this study was to determine if orthotics would reduce postural sway in injured and uninjured subjects. Thirteen subjects with acute inversion ankle sprain and 12 uninjured subjects were assessed for postural sway on the Balance System under two treatment conditions (orthotic and nonorthotic) and four platform movements. A three-factor repeated measures analysis of variance revealed a significant group X treatment interaction, suggesting that postural sway with orthotic intervention improved significantly more in injured subjects than in uninjured subjects. A significant platform movement X treatment interaction, furthermore, revealed that postural sway for the four movement conditions was dependent upon treatment (orthotic vs. no orthotic). Tukey post hoc analysis revealed that orthotic intervention significantly reduced postural sway when the platform moved in the medial/lateral plane and inversion/eversion plane. This research suggests that custom-fit orthotics may restrict undesirable motion at the foot and ankle and enhance joint mechanoreceptors to detect perturbations and provide structural support for detecting and controlling postural sway in ankle-injured subjects. PMID- 8728532 TI - Wobble board training after partial sprains of the lateral ligaments of the ankle: a prospective randomized study. AB - Ankle sprains are often complicated by functional instability and repeated sprains. Rehabilitation with wobble boards in patients with functional instability has been tested, and significant improvement has been found compared to no training. The aim of this study was to investigate whether the number of patients with residual symptoms following ankle sprains could be reduced by training on a wobble board during 12-week recovery period. In addition, the influence of training in the time course reduction of edema was investigated. We performed a prospective study including 61 patients, all active in sports for more than 2 hours a week with primary ankle sprains. The effect of a 12-week training program with wobble board was compared with no training. Forty-eight patients completed the study. In the follow-up period (mean X = 230 days), we found significantly fewer recurrent sprains, and significantly fewer patients in the training group had functional instability of the ankle compared with the no training group. There were no differences in the two groups in the time which elapsed before patients were painless at walking, during running, or at sports. Volumetric measurements revealed no difference in the speed of reduction of hematoma and edema of the ankle and foot between the two groups. We conclude that training on a wobble board early after primary stage 2 ankle sprains is effective in reducing residual symptoms following this lesion and that training does not seem to affect the time course reduction in edema. PMID- 8728533 TI - Recent developments in anxiety, stress, and depression. AB - Recent research in the development, analysis, and pharmacology of animal tests of state anxiety is discussed, including the use of responses to predator odours, the role of learning in modifying the anxiety measured in the plus-maze, and the roles of cholinergic, NMDA, and dopaminergic systems. Developmental and genetic factors are considered with particular reference to the development of tests of trait anxiety. The roles of 5-HT1A receptors in anxiety, depression, impulsivitity, and agonistic behaviours are discussed. Recent studies on the impacts of stress on neurotransmitter, endocrine, and immune systems and the interactions between these systems are discussed, with particular emphasis on their contributions to the development of pathologic states relevant to anxiety and depression. PMID- 8728534 TI - Exploration and predation models of anxiety: evidence from laboratory and wild species. AB - The current article addresses several issues within the context of issues within the context of modeling human anxiety disorders in the laboratory. First, evidence is presented to support the suggestion that behavior in exploration models of anxiety may be motivated by apprehension relating to intraspecific encounters rather than interspecific, predator/prey interactions, which has consequences for the interpretation of findings generated using these tests. Second, data are reviewed concerning the use of stimuli indicating the presence of a predator in the context of anxiety modeling, and it is suggested that tests involving the reactions of animals following exposure to such stimuli may be more closely related to pathologic anxiety mechanisms than tests employing observations during contact with these stimuli. Third, comparative studies, using wild-caught rodents, are outlined that show that, although there are similarities in the defensive strategies adopted by these animals in response to the call of an owl, there are also important differences. Finally, the suggestion is made that the distance-dependent-defense-hierarchy may be of important heuristic value in the interpretation of these data and that, perhaps more significantly, it may also provide a mechanism that allows animal defensive strategies and human anxiety disorders to be placed within the same conceptual framework. PMID- 8728535 TI - A review of the validity and variability of the elevated plus-maze as an animal model of anxiety. AB - Despite or possibly by virtue of the fact that it is one of the most commonly used animal models of anxiety the Elevated Plus-Maze (EPM) results in a wide range of, often contradictory, results following pharmacological experiments. The responses from a questionnaire distributed to 65 groups that have published studies using the EPM in the past 3 years has, along with reference to published reports, enabled some conclusions regarding the influencing factors to be drawn. Some evidence for differential sensitivities between strains exists, with albino rats being more sensitive to the anxiolytic effects of 5-HT3 receptor antagonists and 5-HT1A receptor agonists than pigmented animals. Most important, however, is the manipulation of the animals prior to testing and the aversiveness of the test conditions themselves. Stressing animals before testing (e.g., by moving from holding to test room) or using more aversive test conditions (e.g., elevated light levels) increases sensitivity to potential anxiolytics. Animals that are habituated to gentle handling or tested in less aversive conditions (e.g., EPM with ledges) show reduced likelihood of anxiolytic responses with administration of 5-HT3 antagonists, 5-HT1A agonists, and benzodiazepines. PMID- 8728536 TI - The influence of open arm ledges and maze experience in the elevated plus-maze. AB - In Experiment 1, rats were tested in a plus-maze, with or without small ledges on the open arms, after injection with vehicle or chlordiazepoxide (7.5 mg/kg). They were scored either on their first or second exposure to the maze; those scored on trail 2 had received a 5-min undrugged exposure to the maze 24 h earlier. This dose of chlordiazepoxide had a significant anxiolytic effect on trial 1 only in the maze without ledges, and on trial 2 only in the maze with ledges; thus, the presence of ledges differentially affected anxiolytic sensitivity on trials 1 and 2. The results of a factor analysis study (Experiment 2) confirmed that ledges had a differential effect when rats were repeatedly exposed to the maze. Thus, in the maze without ledges, the scores reflecting anxiolytic activity on trial 1 loaded on one factor, whereas the scores from trials 2 and 3 loaded on another independent factor. In the maze with ledges, the scores reflecting anxiolytic activity on trials 1, 2, and 3 loaded on three independent factors. Considering the published evidence and the results of the present study, we suggest that both types of plus-maze may be measuring the same type of anxiety with different sensitivities on trial 1 (e.g., generalised anxiety or fear of open spaces); different types of anxiety on trial 2 (without ledges--phobia/fear of heights; with ledges--not known), and trial 3 in the maze with ledges, yet another type of anxiety. The factor analysis results are also presented for ethological measures on the plus-maze, and for activity and exploration in the holeboard. Based on the factor loadings, a composite measure of anxiety on trial 1 is presented which will increase the sensitivity of the plus-maze to anxiolytic treatments. The measures of motor activity in the plus-maze load on a different factor from those derived from the holeboard, thus cautioning against considering all measures of motor activity as interchangeable. PMID- 8728537 TI - Plus-maze retest profile in mice: importance of initial stages of trail 1 and response to post-trail cholinergic receptor blockade. AB - Recent research has shown that a single undrugged prior experience of the elevated plus-maze produces significant behavioural changes upon 24-h retest in rats and mice. Typically, when reexposed to the maze, animals display an increased avoidance of the open arms and a corresponding preference for the enclosed sections of the apparatus. Using ethological analyses, the present series of experiments sought to further characterize this phenomenon in mice and to determine whether or not it involves cholinergic receptor mechanisms. Results confirmed that behaviour during Trial 2 is markedly different to that seen on initial exposure, and that such changes are independent of the duration of Trial 1 (2 vs. 5 min). Retest behavioural changes included reduced entry latencies, reduced open arm entries, less time on the open arms and centre platform, lower levels of exploratory head-dipping, and increased entries into and time spent in the closed arms. The importance to the retest phenomenon of the first few minutes of initial exposure was further suggested by min-by-min analyses of the behaviour of animals naive to the maze. Results showed that behaviour during the first min is characterized by high levels of risk assessment from the centre platform and relatively low, but equal, levels of open- and closed-arm exploration. From min 2 onwards, however, behaviour showed a marked change with increasing open arm/centre platform avoidance, increasing closed-arm preference, and decreasing levels of risk assessment and exploratory head-dipping. Thus, it would appear that this within-session aversive learning transfers between sessions to account for behavioural profiles on retest. Irrespective of the duration of Trial 1 (2 or 5 min), posttrial administration of the muscarinic antagonist, scopolamine (0.1 1.0 mg/kg), failed to significantly alter the behavioural changes seen between trials. Data are discussed in relation to the apparent sensitization of fear produced by plus-maze exposure, its possible relation to phobia acquisition, and the need for further research on underlying mechanisms. PMID- 8728538 TI - The pituitary-adrenal axis and the different behavioral effects of buspirone and chlordiazepoxide. AB - Benzodiazepines and the novel anxiolytic buspirone share a common capacity to relieve clinical anxiety but do not share any side effects. Anxiety releases stress hormones and, at moderate doses, anxiolytic benzodiazepines block this release. It is interesting, therefore, that buspirone and other 5-HT1A agonists release stress hormones at moderate doses. Both the U-shaped dose-response curve seen with buspirone in some animal tests of anxiety and its slow onset of clinical action could be attributed to this release of stress hormones. Metyrapone (200 mg/kg), an inhibitor of 11-beta-hydroxylase, was used in the present experiments as a form of chemical adrenalectomy and was combined with administration of corticosterone (1 mg) to produce rats with presumed approximately normal corticosterone levels but no capacity to release endogenous corticosterone. This treatment reduced the difference normally observed in the effects of chlordiazepoxide (5 mg/kg) and buspirone (0.37 mg/kg) on a fixed interval schedule particularly in the early part of the interval when release of behavioral inhibition would be expected to contribute most to the effects. These results are consistent with the previous suggestion of Johnston and File (8) that the anxiolytic action of buspirone may be counteracted by activation of the pituitary-adrenal axis. Corticosterone appears to be the most likely critical agent for this antagonist action in the present experiments, although CRF and ACTH are also possibilities. It is likely that there is a mutual functional opposition between endogenous anxiolytic factors and stress hormones. PMID- 8728539 TI - Muscarinic antagonists are anxiogenic in rats tested in the black-white box. AB - Central cholinergic (ACh) projections have been shown to modulate stress-induced activation of the hypothalamic-pituitary-adrenal (HPA) axis and are integral to the expression of electrophysiological correlates of arousal, namely hippocampal theta rhythm. The degree to which these actions of ACh are behaviorally relevant has received comparatively less attention, and we sought to investigate if manipulations of ACh systems might also affect behaviors related to stress and arousal. We chose to examine indices of anxiety as revealed by changes in behavior elicited by the black-white box test, a relatively novel and recently validated model of rodent anxiety. Groups of rats were injected with either scopolamine hydrobromide (SCOP; 0, 0.05, and 0.10 mg/kg i.p.) or the peripherally acting scopolamine methyl bromide (methyl-SCOP; 0, 0.05, and 0.10 mg/kg i.p.) to compare and contrast the effects of central and peripheral ACh blockade on measures of anxiety. SCOP pretreatment significantly lowered latencies for rats to escape from the white to black compartment, while methyl-SCOP elevated latencies to reenter the white chamber from the black. Both drugs increased the amount of time rats spent in the black compartment and also suppressed exploration as revealed by decreased episodes of intercompartmental locomotion. Neither drug deleteriously affected locomotor activity, however; in fact, SCOP significantly increased locomotion in the white chamber. In the absence of motor disturbances to account for any group differences, we contend that both central and peripheral ACh blockade may affect measures of anxiety, perhaps by directly or indirectly affecting HPA activity. Central ACh systems may underlie sensory filtering whereby irrelevant stimuli are excluded from sensory processing. Antagonism of ACh transmission may render an animal incapable of correctly processing sensory information leading to hyperresponsiveness, which can manifest itself as enhanced anxiety and fear. PMID- 8728540 TI - NGF regulatory role in stress and coping of rodents and humans. AB - Nerve growth factor (NGF) is a polypeptide growth factor which exerts trophic and differentiative effects on specific peripheral and central populations of neurons. Recent data showing that various cellular types of the endocrine and immune systems are able to synthesize and release NGF have suggested that this neurotrophic factor may also play an important role in vertebrate physiologic homeostasis. Previous studies using a mouse model of aggressive behavior have shown that NGF levels increase in both plasma and the CNS following intermale agonistic encounters. More recently, we have extended this research area to include other species: in particular, humans. The data now available indicate that labour and lactation, or the occurrence of a stressful event such as the very first jump with a parachute causes in NGF plasma levels as well as changes in the distribution of NGF receptors on lymphocytes. This review aimed to outline the current understanding of NGF role in vertebrates in stress-related events. PMID- 8728542 TI - Modulation of plus-maze behaviour in mice by the preferential D3-receptor agonist 7-OH-DPAT. AB - Differences in the behavioural profiles of dopamine D2 receptor antagonists (e.g., haloperidol vs. sulpiride) in a animal models of anxiety have prompted speculation concerning the importance of their relative affinities for D2-like receptor populations. In an initial attempt to investigate the involvement of D3 receptors in anxiety, the present study examined the effects of the preferential D3-receptor agonist, (+/-)7-OH-DPAT (0.01-10.0 mg/kg), on behaviours displayed by male mice in the elevated plus-maze paradigm. An ethological approach incorporating measurement of a range of defensive acts and postures in addition to conventional parameters was used to provide a comprehensive behavioural profile for the compound. Data analysis indicated a significant increase in percentage of open-arm entries at 10 mg/kg and an altered temporal distribution of behaviour at 1-10 mg/kg. Furthermore, risk-assessment measures (stretched attend postures, closed-arm returns) were dose dependently reduced by drug treatment. Although these behavioural changes would be consistent with anxiety reduction, such an interpretation is negated by dose-dependent decreases in all active behaviours (arm entries, rearing, and head-dipping) and by marked increases in entry latencies and nonexploratory behaviour at the highest dose tested. Overall, these effects are remarkably similar to those previously reported for quinpirole, suggesting either that D2 and D3 receptors exert similar behavioural control or that the agents employed are sufficiently potent at D2 receptors to prevent a resolution of D2 and D3 responses. PMID- 8728541 TI - Anticonflict effects of a competitive NMDA receptor antagonist and a partial agonist at strychnine-insensitive glycine receptors. AB - Using the conflict drinking Vogel test in rats as a model, in the present study we examined the anxiolytic-like activity of DL-(E)-2-amino-4-methyl-5-phosphono-3 pentenoic acid (CGP 37849), a competitive N-methyl-D-aspartate (NMDA) receptor antagonist and 1-aminocyclopropanecarboxylic acid (ACPC), a partial agonist at strychnine-insensitive glycine receptors associated with the NMDA receptor complex, after their intraperitoneal (i.p.) and intrahippocampal (IHP) administration. CGP 37849, administered in doses of 1.25-5 mg/kg i.p., produced an anticonflict effect in a dose-dependent manner, but was inactive when injected in doses of 0.01-0.1 micrograms IHP. At the same time, when administered in higher doses (10 mg/kg i.p. or 0.3 micrograms IHP), that drug induced motor impairment. On the other hand, ACPC exhibited an anxiolytic-like activity after both i.p. (100-200 mg/kg) and IHP (3-30 micrograms) administration. These results, as well as the literature data on the lack of motor-impairing effects of ACPC, indicate that the latter drug seems to be more advantageous than CGP 37849 as a potential therapeutic agent in the treatment of anxiety disorders. Furthermore, they also show that the hippocampus may be one of the neuroanatomical sites of the anxiolytic-like effect of ACPC, but not of CGP 37849. PMID- 8728543 TI - Behavioural and neurochemical responses of male and female chicks to cat odour. AB - In the first experiment male chicks were exposed to neutral and cat odours at days 4, 7, or 10 after hatching. Of the chicks tested at day 4, few made contact with either odour cloth, but those tested at day 7 made fewer contacts with the cat odour cloth, compared with the neutral odour, spent less time in contact with it, and spent more time in the zone furthest from the cloth. These clear differences were not seen in the group tested at day 10. In a second experiment, the behaviour of day 7 male chicks was compared in the presence of neutral, disinfectant, chick blood or cat odours, and the most extreme differences were between neutral and cat odours. In a third experiment, both male and female chicks were exposed to cat odour at day 7 and both showed similar avoidance. After exposure to cat odour both sexes showed significantly reduced GABA enhancement of benzodiazepine binding; which is a change associated with increased fear. However, after exposure to cat odour, they also showed significant decreases in 5-HT availability evidenced by lower basal and K(+) evoked [3H]-5-HT release and, in the male chicks only, by an increased [3H]-5-HT uptake from archistriatal slices. These changes in 5-HT function are in the direction associated with reduced fear and would, therefore, seem to be adaptive and compensatory in function. Neither male nor female chicks showed any differences in [14C]-GABA release or uptake as a result of exposure to cat odour. Although the pattern of response to cat odour was the same in both male and female chicks at day 7, there were significant sex differences in 5-HT and GABA tone and benzodiazepine binding; these sex differences were also found in day 10 chicks. The importance of these for sex differences in trait anxiety is discussed. PMID- 8728544 TI - Relationship between memory and fear: developmental and pharmacological studies. AB - Habituation of the acoustic startle response (ASR) and freezing responses were assessed simultaneously in rats of different ages. Results showed that until 30 days of age rats were not able to express long-term habituation of the ASR, whereas irrespective of age, all rats exhibited the increased freezing responses as a result of fear conditioning. In addition, the interaction between fear and memory was studied using the same behavioral model in adult rats treated intraperitoneally with diazepam at doses 1.2 and 2.5 mg/kg. Diazepam administration did not result in a significant change in initial startle responsiveness but elicited a profound suppression of startle amplitude over trials. Furthermore, animals given diazepam showed more initial freezing than the vehicle-treated controls, while fear-conditioned freezing was decreased by diazepam. The pattern of results is discussed in relation to developmental and pharmacological dissociations between the different behavioral components of responses to aversive and stressful cues. PMID- 8728545 TI - Affiliation in periadolescent rats: behavioral and corticosterone response to social reunion with familiar or unfamiliar partners. AB - The aim of this study was to investigate whether the social relationship between periadolescent rats, which is characterized by a high expression of play behavior, resembles attachment or whether it reflects a nonspecific social attraction between peers. Sprague-Dawley rats have been kept in pairs with a conspecific of the same age and sex from the time of weaning (PND 21). On PND 34, the members of each pair were separated for a 24-h period and randomly assigned to one of three groups: a) immediately sacrificed (NT = nontreated); b) reunited for 30 min with their previous cagemate (FAM); or c) with an unfamiliar conspecific of the same age and sex (UNF). During the reunion, the occurrence of both social and nonsocial behaviors was scored. Overall, periadolescent UNF animals spent more time in social investigation than FAM rats, which were conversely more involved in cage-oriented activities. In addition, higher levels of rough-and-tumble play were expressed in encounters between UNF subjects than between FAM rats. Finally, within the female group, UNF rats were more involved in play soliciting and less in digging activity than FAM animals. Blood corticosterone levels, which were measured at the end of the separation period, were higher in females than in males and were significantly lower than following reunion in a novel environment. The degree of familiarity did not affect hormonal changes. Results suggest that periadolescent rats do not form attachment relationships with each other. However, partner familiarity appears to be an important variable affecting social relationships. PMID- 8728547 TI - Behavioral stress response of genetically selected aggressive and nonaggressive wild house mice in the shock-probe/defensive burying test. AB - Genetically selected aggressive and nonaggressive male wild house mice were tested in the shock-probe/defensive burying test: Five distinct behaviors (burying, immobility, rearing, grooming, and exploration) were recorded in two environmental situations: fresh and home cage sawdust. Nonaggressive animals, characterized by a Long Attack Latency (LAL), showed more immobility in both test situations than animals having Short Attack Latencies (SAL), whereas SAL males displayed more defensive burying than LAL ones when tested with fresh sawdust. Testing with home cage sawdust, however, resulted in the same duration of defensive burying in SAL and LAL. These results support earlier findings about the existence of two heritable, fundamentally different strategies to cope with aversive situations. Aggressive (SAL) animal react actively to environmental challenges, whereas nonaggressive animals react actively or passively, depending on the characteristics of the stressful environment. These mouse lines, selected for attack latency, i.e., aggression, may, therefore, be important tools to unravel the genetic architecture underlying the physiological and neuronal mechanisms of behavioral strategies towards stressful events. PMID- 8728546 TI - Strain differences in fear-motivated behavior of rats. AB - Studies have shown different and sometimes contradictory results in response to anxiolytic drugs. In the present study, the behavioral performance of rat strains, obtained from different breeders, was examined in four animal models of anxiety- or in exploration-related behavior to assess the potential contribution of genetic disposition or breeding factors to aversion-motivated behavior. Male rats: Wistar/Winkelmann, Wistar/Charles River, Wistar/BGVV, Lewis/Charles River, Fischer/Charles River, Brown Norway/Charles River were used in a conflict test in the open field, a free exploratory paradigm, social interaction test, and the holeboard test. The results show that robust behavioral differences in anxiety or exploration exist between different strains of rats and animals of one strain, obtained from different breeders. The differences shown in anxiety-related behavior might explain sometimes contradictory effects following the treatment with anxiolytic or anxiogenic drugs. The results indicate that genetic factors and breeding conditions substantially contribute to anxiety-motivated behaviors in animal models of anxiety. These differences in anxiety-related behavior may also be related to biochemical differences. PMID- 8728548 TI - Differences in benzodiazepine binding in quail selectively bred for differences in tonic immobility. AB - We have previously found that quail selectively bred to exhibit long (LTI) or short (STI) tonic immobility responses to manual restraint differed with respect to the affinity of binding at the diazepam-sensitive benzodiazepine binding site. In the current study, binding at other components of the GABAA-benzodiazepine receptor complex was investigated. Whereas the lines did not differ in either number or affinity of GABAA receptors, we found that GABA caused significantly greater enhancement of [3H]flunitrazepam binding in the forebrains of LTI than in STI birds. There was also significantly higher binding to the diazepam insensitive component (alcohol binding site) of the GABAA-benzodiazepine complex in the forebrain of the LTI line. It is not known, however, whether this difference in the lines is due to differences in number or affinity of these sites. It is discussed whether these manifold differences between the lines in binding at the forebrain sites of the GABAA-benzodiazepine complex could be founded on subunit differences, and whether differences in benzodiazepine binding could underlie the genetically determined behavioural differences in tonic immobility. PMID- 8728549 TI - Anxiolytic effects in the plus-maze of 5-HT1A-receptor ligands in dorsal raphe and ventral hippocampus. AB - The response of rats naive to, or experienced with, the elevated plus-maze test of anxiety was observed following direct administration of the 5-HT1A-receptor agonist (+/-)-8-hydroxy-dipropylaminotetralin (8-OH-DPAT) (50, 100, or 200 ng) or antagonist tertatolol (3 micrograms) into the dorsal raphe nucleus or bilaterally into the ventral hippocampus. In rats naive to the plus-maze, neither drug had a significant effect when microinjected into the dorsal raphe nucleus. However, in rats experienced with the plus-maze, 8-OH-DPAT (100 and 200 ng) had significant anxiolytic effects when administered to the dorsal raphe nucleus, which were antagonised by tertatolol (3 micrograms); this suggests they were mediated by 5 HT1A receptors. Hyperactivity (increased number of closed-arm entries) was found following bilateral injection of 8-OH-DPAT (100 ng) into the ventral hippocampus of rats naive to the plus-maze. This was not completely antagonised by tertatolol (3 micrograms). Interestingly, tertatolol (3 micrograms) itself had an anxiolytic effect which was not antagonised by 8-OH-DPAT (100 ng), suggesting the effect was not mediated by 5-HT1A receptors, and indeed other actions of tertatolol, such as those on 5-HT1B or beta-adrenergic receptors could have been involved. In rats experienced with the plus-maze, tertatolol (3 micrograms) again had a significant anxiolytic effect when administered bilaterally to the ventral hippocampus, and again, this was not antagonised by 8-OH-DPAT (100 ng). These results demonstrate that both the intracerebral location of the injection and test experience profoundly influence the effects of 5-HT1A ligands on behaviour of rats in the elevated plus-maze test of anxiety. PMID- 8728550 TI - Role of 5-HT in stress, anxiety, and depression. AB - There are conflicting results on the function of 5-HT in anxiety and depression. To reconcile this evidence, Deakin and Graeff have suggested that the ascending 5 HT pathway that originates in the dorsal raphe nucleus (DRN) and innervates the amygdala and frontal cortex facilitates conditioned fear, while the DRN periventricular pathway innervating the periventricular and periaqueductal gray matter inhibits inborn fight/flight reactions to impending danger, pain, or asphyxia. To study the role of the DRN 5-HT system in anxiety, we microinjected 8 OH-DPAT into the DRN to inhibit 5-HT release. This treatment impaired inhibitory avoidance (conditioned fear) without affecting one-way escape (unconditioned fear) in the elevated T-maze, a new animal model of anxiety. We also applied three drug treatments that increase 5-HT release from DRN terminals: 1) intra-DRN microinjection of the benzodiazepine inverse agonist FG 4172, 2) intra-DRN microinjection of the excitatory amino acid kainic acid, and 3) intraperitoneal injection of the 5-HT releaser and uptake blocker D-fenfluramine. All treatments enhanced inhibitory avoidance in T-maze. D-Fenfluramine and intra-DRN kainate also decreased one-way escape. In healthy volunteers, D-fenfluramine and the 5-HT agonist mCPP (mainly 5-HT2C) increased, while the antagonists ritanserin (5 HT2A/2C) and SR 46349B (5-HT2A) decreased skin conductance responses to an aversively conditioned stimulus (tone). In addition, D-fenfluramine decreased, whereas ritanserin increased subjective anxiety induced by simulated public speaking, thought to represent unconditioned anxiety. Overall, these results are compatible with the above hypothesis. Deakin and Graeff have suggested that the pathway connecting the median raphe nucleus (MRN) to the dorsal hippocampus promotes resistance to chronic, unavoidable stress. In the present study, we found that 24 h after electrolytic lesion of the rat MRN glandular gastric ulcers occurred, and the immune response to the mitogen concanavalin A was depressed. Seven days after the same lesion, the ulcerogenic effect of restraint was enhanced. Microinjection of 8-OH-DPAT, the nonselective agonist 5-MeO-DMT, or the 5-HT uptake inhibitor zimelidine into the dorsal hippocampus immediately after 2 h of restraint reversed the deficits of open arm exploration in the elevated plus maze, measured 24 h after restraint. The effect of the two last drugs was antagonized by WAY-100135, a selective 5-HT1A receptor antagonist. These results are compatible with the hypothesis that the MRN-dorsal hippocampus 5-HT system attenuates stress by facilitation of hippocampal 5-HT1A-mediated neurotransmission. Clinical implications of these results are discussed, especially with regard to panic disorder and depression. PMID- 8728551 TI - Role of 5-HT1A receptors in the effects of acute chronic fluoxetine on extracellular serotonin in the frontal cortex. AB - Fluoxetine 10 mg/kg i.p. significantly increased the extracellular concentrations of serotonin (5-HT) in the frontal cortex as assessed by in vivo microdialysis. This effect was significantly potentiated when 0.3 mg/kg s.c. WAY-100635, a 5 HT1A receptor antagonist, was administered 30 min before. WAY-100635 by itself had no effect on extracellular 5-HT. Twenty-four hours after chronic fluoxetine schedule (10 mg/kg/day i.p. x 14 days), basal extracellular 5-HT concentrations in the frontal cortex were higher than those of animals that had received the vehicle chronically. At 24 h after the last dose, a challenge dose of fluoxetine (10 mg/kg i.p.) raised extracellular 5-HT similarly in chronically vehicle or fluoxetine treated rats. At this same interval 25 micrograms/kg s.c. 8-OH-DPAT, a 5-HT1A receptor agonist, significantly reduced extracellular 5-HT only in the frontal cortex of rats treated chronically with the vehicle. Examining basal extracellular 5-HT, the effect of a challenge dose of fluoxetine and the effect of 25 micrograms/kg 8-OH-DPAT after 96 h washout, no differences were found between chronically fluoxetine and vehicle-treated rats. The results confirm that the ability of fluoxetine to stimulate 5-HT1A autoreceptors through an increase of endogenous 5-HT attenuates its effect on cortical dialysate 5-HT. Chronic fluoxetine increased the basal concentrations of extracellular 5-HT only when a substantial amount of its metabolite was present in the brain and during the desensitization of presynaptic 5-HT1A autoreceptors (24 h after the last dose). These effects, in fact, disappeared after 96 h washout. The continuous presence of the drug may, therefore, be necessary to maintain extracellular 5-HT at concentrations high enough to produce a therapeutic effect. PMID- 8728552 TI - Effects of psychotropic drugs on rat responding in an operant paradigm involving choice between delayed reinforcers. AB - Preference for immediate reward, taken as an index of impulsiveness, has been suggested to be under the preferential control of central serotonin (5-HT) function. The present study examined the effects of the acute administration of drugs which directly or indirectly alter 5-HT transmission on tolerance to delay of reward in rats subjected to a procedure of discrete-trial choice in an operant chamber. Different groups of rats were trained to choose between two levers giving access to reinforcers differing in both magnitude and delay: one food pellet, delayed by 0 or 5 s, vs. five pellets delivered after a prereinforcer interval fixed at either 15, 30, 45, or 60 s, depending on the experiments. The learning curves indicated that rats were able to adjust their choice strategy precisely according to various factors: the respective duration of the delays before the small and large rewards, the immediacy of the small reward delivery, and the lengthening of the trials by a postreinforcer delay (or intertrial interval). Whatever the experimental parameters and stage of the learning, an acute administration of drugs able to reduce 5-HT neuronal activity (benzodiazepines; 5-HT1A receptor partial agonists: buspirone and MDL 73005EF) or enhance 5-HT transmission (5-HT reuptake inhibitors: indalpine and zimelidine; 5 HT1A receptor full agonist: 8-OH-DPAT) failed significantly to alter choice strategy (decreased or increased preference for the large but delayed reward, respectively), as they did in other situations such as a T-maze procedure. Only d amphetamine (0.5 mg/kg), on one occasion, significantly reduced preference for the larger reward. The choice strategy was also insensitive to acute changes in experimental parameters such as a reduction in delay or increase in the magnitude of the large reinforcement. These results indicate that the present operant paradigm is not sensitive to acute modifications in the internal state of the animals and in the reward contingencies, and therefore is not useful to evaluate tolerance to delay and variations in impulsiveness in rats. PMID- 8728553 TI - Effects of the 5-HT1A antagonist (+)-WAY-100135 on murine social and agonistic behavior. AB - Compounds previously identified as 5-HT1A antagonists have subsequently been demonstrated to possess partial agonistic properties in models assessing somatodendritic autoreceptor function. This study examined the influences of (+) WAY-100135, claimed to be the first selective 5-HT1A antagonist, on offensive behaviour in male mice. Employing a resident-intruder paradigm, administration of (+)-WAY-100135 (1.0-10.0 mg/kg s.c.) enhanced elements of resident offensive behaviour at 2.5 and 5.0 mg/kg but reduced such behaviour at 10.0 mg/kg. In comparison, resident defensive postures remained unchanged except for a significant increase in defensive sideways behaviour at 10.0 mg/kg. These effects were accompanied by reduced rearing behaviour across the dose range tested. Attend/approach behaviour was significantly reduced at the lowest, but increased at the highest, doses tested. Such results may reflect response competition rather than concomitant motor impairment. Given the dynamic behavioural interactions occurring in this paradigm, the increased offensive behaviour of the resident mice leads to enhanced defence and counter-attack by the intruder conspecifics. The results are discussed with reference to the current literature concerning the behavioural effects of other 5-HT1A antagonists. PMID- 8728554 TI - Selective effects of 8-OH-DPAT on social competition in the rat. AB - Previous research has demonstrated that dominant-subordinate relationships measured in small groups of rats competing for access to palatable food or fluids can be disrupted by both anxiolytic and anxiogenic drugs, and it has been proposed as a possible animal model of anxiety. The present study investigated the effects of the selective 5-HT1A agonist 8-OH-DPAT on the rank order of triads of rats measured in terms of access to sweetened milk. The effect of 8-OH-DPAT on locomotor activity and intake of sweetened milk was also determined. 8-OH-DPAT (25 and 37.5 micrograms/kg) significantly increased the subordinate animals position in the social hierarchy without effect on the individual intakes of sweetened milk or locomotor activity. The same doses administered to dominant animals had no effect on any of the parameters measured. The 8-OH-DPAT-induced increase in social competition in subordinate rats was dissociable from effects on feeding behavior and locomotor activity. The results from this study provide further evidence that social competition in groups of rats may represent a model that can be used to detect drugs acting via receptor mechanisms believed to be implicated in anxiety. PMID- 8728555 TI - Effects of chlordiazepoxide on maternal aggression in mice depend on experience of resident and sex of intruder. AB - Lactating mice respond differentially to intruders of differing sex, displaying defensive attack against the male and offensive attack against the female. Such a phenotypic dichotomy appears to have adaptive value in that unfamiliar males pose a much greater threat to the offspring than do females. The present study examined the effects of the benzodiazepine anxiolytic chlordiazepoxide (CDP) (2.5 10.0 mg/kg) on this differential response pattern in aggression-naive (nonscreened) (NS) and aggression-experienced (screened) (S) lactating female mice (Mus musculus domesticus) confronting intruders of either sex in a 10-min test. This procedure was used to evaluate the influence of both the type of opponent and previous aggressive experience on basal behavioural profiles and drug action. Results showed that both intruder sex and prior screening for attack modulated the behaviour of lactating females toward intruders. In turn, both variables strongly influenced CDP effects on maternal aggression. In particular, in S dams, CDP dose-dependently increased maternal attack against males but decreased attack against female intruders. Conversely, in NS dams, CDP decreased attack (and fear) against males but did not affect it against females. In both S and NS conditions, CDP modified the attack strategy of lactating females against the male, switching it from a defensive to an offensive pattern. Exploration, social investigation, eating, and immobility were differentially affected by the drug treatment, depending on screening and/or intruder sex condition. These differential effects of CDP between S and NS conditions, toward either male or female intruders, cannot be fully explained by differences in the baseline levels of these behaviours. Alternative hypotheses are discussed. These findings demonstrate that the effects of CDP on maternal attack behaviour depend on not only the drug but also the object of attack, and hence the function of attack and the prior experience of the attacker. PMID- 8728556 TI - Prenatal naltrexone facilitates male sexual behavior in the rat. AB - The involvement of endogenous opiates in the differentiation of sexual behavior was tested by exposing rat fetuses to continuous naltrexone during the last 9 days of gestation. Time-mated female rats received oral naltrexone, 40 mg/kg/day, via their drinking water, from gestational day 13 until parturition. Early motor development, measured by swimming ability in 7-, 9-, and 11-day-old offspring of the treated dams, was unaffected by prenatal naltrexone. Adult male offspring were given three tests of male sexual behavior, then castrated, primed with ovarian hormones, and given two tests of feminine receptivity (lordosis quotient). Prenatal naltrexone facilitated masculine behavior and suppressed feminine receptivity: latencies to first mount and to ejaculation were shorter, mount rate was higher, and lordosis quotient was lower in naltrexone-treated rats, compared with control animals. These findings implicate endogenous opiates in prenatal organization of sex-specific behavioral dispositions. PMID- 8728557 TI - Involvement of the 5-HT1A subtype receptor in the neonatal organization of agonistic behaviour in the rat. AB - 5-hydroxytryptamine (5-HT) interacts with testosterone (T) in the development of a number of neuronal systems controlling sexually dimorphic adult behaviours. In this report, we investigated this interaction on the organization of agonistic behaviour in males, females, androgenized females (250 micrograms/pup of T proprionate on the day of birth), and males castrated on the day of birth. We have shown previously that manipulating 5-HT2 activity over the 2nd week of life modulates adult agonistic behaviour, depending on genetic sex and the presence of T. In this report, we investigated the effects seen in adulthood of a 5-HT1A agonist [8-OH-DPAT, 0.25 mg/kg, intraperitoneally (i.p.)] and antagonist (WAY100135, 0.25 mg/kg, i.p.) given over days 8-16 postpartum. The test for agonistic behaviour was carried out in a neutral territory against a matched conspecific, and introductory, offensive and defensive activities were note. Results show that neonatal administration of the 5-HT1A antagonist WAY100135 increases introductory activity and defense in the presence of neonatal T, independent of genetic sex, because these effects were seen in sham-castrated males and androgenized females. Offence followed a similar pattern, in that it was increased by WAY100135, but only in males. In the case of defence, the effects of the antagonist were reinforced by the action of the agonist (8-OH DPAT) in both males and females, indicating an inhibitory role of 5-HT1A perinatal activity on defence in the presence of malelike levels of circulating T and a facilitatory role when levels of T are low or negligible. These findings indicate that 5-HT1A activity is involved in the development of agonistic behaviour and the effects are influenced by T. The results also show that the offensive and defensive facets of agonistic activity are controlled differently. PMID- 8728558 TI - Neonatal organizational effects of the 5-HT2 and 5-HT1A subsystems on adult behavior in the rat. AB - Males, females, neonatally androgenized females, and neonatally castrated males were treated over the second week of life with 0.25 mg/kg of either the 5-HT2 agonist 1-(2,5-dimethoxy-3-iodophenyl)-2-aminopropane HCl (DOI), the 5-HT2 antagonist ritanserin (Rit), the 5-HT1A agonist 8-hydroxy-2-(di-n propylamino)tetralin (8-OH-DPAT), or the 5-HT1A antagonist WAY100135 (WAY). Exploration, anxiety, sociosexual preferences, and sexual behavior were measured in adulthood. Agents acting on 5-HT1A receptors do not appear to affect organization of any of the behavioral systems studied. DOI increased exploratory activity but in females only, which suggests that testosterone antagonizes the stimulatory effect of 5-HT2 activity on exploration. Neonatal ritanserin selectively reduced anxiety in females, and DOI had a similar effect in androgenized females. This indicates that neonatal 5-HT2 activity is anxiogenic in normal females, anxiolytic in androgenized females, and has no effect on anxiety in males. Males and androgenized females both showed a preference for the female teaser that was abolished by the 5-HT2 agonist, DOI. These results point out that 5-HT2 activity selectively suppresses heterosexual preference induced in the presence of neonatal testosterone. DOI also reduced both male sexual behavior in males and female sexual behavior in androgenized females. Thus, the 5-HT2 system antagonizes the action of testosterone in stimulating heterosexual orientation and sexual activity, and this is independent of genetic sex. PMID- 8728559 TI - Stress-induced increase in brain neuroactive steroids: antagonism by abecarnil. AB - Acute foot shock stress elicits a selective and time-dependent increase of neuroactive steroid (pregnenolone, progesterone, allotetrahydrodeoxycorticosterone) concentrations in rat brain cortex, accompanied by a marked increase of plasma corticosterone. The brain cortical neuroactive steroid levels peaked between 10 and 30 min poststress and returned to control values by 2 h. Abecarnil (0.3 mg/kg), i.p.), a beta-carboline derivative with anxiolytic properties, completely antagonized the effect of foot shock on brain cortical neuroactive steroids. A single administration of the anxiogenic beta-carboline FG 7142 (15 mg/kg, i.p.), in contrast, mimicked the effect of foot shock. These data support the hypothesis for the existence of a functional relationship between brain neuroactive steroid concentrations and GABAA receptor function/emotional state of the animal. PMID- 8728560 TI - Stress: a major variable in the psychopharmacologic response. AB - The role of central monoaminergic neurones in stress is undisputed, albeit undefined. This is partly because little is known about the influence of the type or intensity of stress, or subjects' stress history, on monoaminergic transmission. That the presynaptic response is stimulus specific is underlined by a study using in vivo microdialysis in freely moving rats. This indicated that graded changes in noradrenaline efflux in the frontal cortex are produced by progressively increasing the number of novel features in the rats' environment. The influence of receptor status on behavioural responses to stress also depends on the stress imposed. This was suggested by studies showing that rats' behavioural response to stress correlated with the density of cortical beta adrenoceptors. But the precise relationship again depended on features of the stress, possibly its intensity. Finally, it seems that even a single stress challenge (a 6-min swim) causes a long-latency increase in the density of 5-HT2A receptors in mouse cortex. This upregulation was prevented by a history of intraperitoneal injections of saline but not by injections of the monoamine reuptake blocker sibutramine hydrochloride. Collectively, these experiments emphasize the importance of stress as an experimental variable when studying the actions of psychotropic drugs. PMID- 8728561 TI - Psychosocial stress in tree shrews: clomipramine counteracts behavioral and endocrine changes. AB - Male tree shrews (Tupaia belangeri) provide an animal model to study the neurobehavioral consequences of chronic psychosocial stress. When living in visual and olfactory contact with a male conspecific by which it has been defeated, the subordinante tree shrew shows dramatic behavioral, physiological, and neuroendocrine changes. Because the over all pattern of these changes resemble a depression-like symptomatology, we investigated to what extent the behavioral and endocrine changes in subordinate animals can be reversed by treatment with the tricyclic antidepressant clomipramine. In the present study, animals were subjected to a 10-day period of psychosocial conflict to elicit stress-induced behavioral and endocrine alterations before the onset of drug treatment, and psychosocial stress continued throughout the treatment period of 30 days. Clomipramine was administered orally once daily at a dose of 50 mg/kg. The drug had a time-dependent restorative influence on marking and grooming behavior, locomotor activity, risk assessment, as well as on urinary cortisol and norepinephrine excretion. It, thus, appears that the clomipramine treatment counteracts the behavioral and endocrine effects of chronic psychosocial stress in tree shrews, and the time course of recovery corresponds closely to that observed when treating depressed patients in the clinic. PMID- 8728562 TI - Effect of chronic mild stress on circadian rhythms in the locomotor activity in rats. AB - The purpose of this study was to assess whether the chronic mild stress (CMS) procedure, as a realistic animal model of depression, affects the rhythms of the locomotor activity in rats. Rhythm parameters (period, mesor, amplitude, acrophase, and percent rhythm) were estimated from the best-fitted cosine function curves. Period is the length, mesor is the mean level, amplitude (A) is the extent, acrophase is the timing of the rhythm; percent rhythm represents the variability estimated by the cosine regression and expressed as a percentage of the total variability of raw data. The animals were kept on the 12 L : 12 D cycle during 13 weeks of the experiment and subjected to CMS for first 4 weeks. In week 5 the rats were under the constant light for 24 h a day (LL), and in week 9, under the constant darkness (DD). In LD 12:12 CMS decreased the activity in the dark phase by approximately 50% (p < 0.01) and did not change the activity in the light phase, resulting in a drop of the 24 h activity by about 40% in comparison to controls. The amplitude of diurnal variations of the activity was highly statistically different from zero at p(A = 0) < 0.0001, and the percent rhythm was in range of 40-75% in both the CMS and control groups. The mesor and the amplitude of the diurnal rhythm (with a period of 24 h) in the CMS rats were significantly (p < 0.001) lower than those in the control. In LL, the activity of both groups was diminished about 50% during the subjective dark phase. On the other hand, in the subjective light phase the activity of CMS rats only was diminished. The percent rhythm for the CMS and control rats was 30 and 58%, respectively, and values of mesor, amplitude, and acrophase for both groups were highly statistically different. In DD, the activity in the CMS group was statistically significantly lower in both the subjective dark and light phases. In contrast to the results from LL, the cosine curves from DD were similarly shifted in relation to the subjective light-dark cycle. After a restoration of the LD cycle the levels of the 24-h activity of both groups became equal in the 13th week, but the light and dark phase differences between the groups were still statistically significant (p < 0.05). The present results indicate that CMS exerts distinct and prolonged disturbances of the diurnal and circadian rhythms of the locomotor activity in the rats. PMID- 8728563 TI - Indirect dopamine agonists effects on despair test: dissociation from hyperactivity. AB - Both dexamphetamine and the pure dopamine reuptake inhibitor GBR 12783 elicit a stimulation of locomotion and increase swimming activity in the behavioral despair test in mice. The dopamine D1 dopamine receptor antagonist SCH 23390 dose dependently (7.5-30 micrograms/kg s.c.) antagonized the stimulant locomotor effect on both drugs but did not prevent their antiimmobility effect on the behavioral despair test. The D2 dopamine receptor antagonist haloperidol dose dependently (12.5-50 micrograms/kg i.p.) antagonized the effects of dexamphetamine on both locomotor activity and behavioral despair test. By contrast, haloperidol inhibited the effects of GBR 12783 in the forced swimming test but not on locomotion. It is concluded that indirect dopamine agonists are effective on the behavioral despair test independently of a stimulation of locomotor activity. Their effects on the despair test depend on the stimulation of D2 but not D1 dopamine receptors. PMID- 8728564 TI - Expression of Fos protein in various rat brain areas following acute nicotine and diazepam. AB - We studied the effects of an acute dose of (-)-nicotine (1 mg/kg) on Fos-like immunostaining (IS) in rat brain areas. Nicotine increased Fos IS significantly in the medial terminal nucleus of accessory optic tract (MT), and tended to increase it in the interpeduncular nucleus (i.p.), as well as in the stress related areas, the paraventricular hypothalamic nucleus (PVN) and the supraoptic nucleus (SON). Previously nicotine was reported to increase Fos IS also in another stress-related area, the central nucleus of amygdala (ACe). This led us to study the interaction of nicotine with diazepam (10 mg/kg). Diazepam alone increased Fos IS in PVN and in SON as well as in ACe. In diazepam- and nicotine treated rats Fos IS was increased in PVN and SON as well as in MT and i.p.. In MT and i.p. of diazepam and nicotine-treated rats Fos IS was similar to that induced by nicotine alone, and in PVN and SON of these rats Fos IS in ACe. Taken together, diazepam induced Fos IS in all stress-related areas studied (PVN, SON, ACe), but not in central visual structures, where nicotine induces Fos IS (MT, i.p.). No significant interactions on Fos expression were found between acute effects of diazepam and nicotine suggesting that these drugs activate different sets of neurons within the stress-related brain areas. PMID- 8728565 TI - Neuroendocrine effects of diazepam and flesinoxan in the stress-induced hyperthermia test in mice. AB - In the stress-induced hyperthermia (SIH) paradigm in mice, both a benzodiazepine receptor agonist, diazepam, and a 5-HT1A receptor agonist, flesinoxan, reduced the stress-induced increase in rectal temperature. The SIH procedure itself enhanced plasma ACTH and corticosterone levels but not plasma glucose levels. Diazepam (3, 6, and 12 mg/kg p.o.) did neither affect basal plasma ACTH, corticosterone, or glucose levels, nor did it suppress the stress-induced rises in these parameters. Flesinoxan (1, 3, and 10 mg/kg p.o.) enhanced plasma ACTH and corticosterone concentrations under nonstress conditions but did not affect the stress-induced increases in ACTH and corticosterone secretion. No clear effects of flesinoxan on plasma glucose levels were found. Our results indicate that in mice the anxiolytic effects of diazepam and flesinoxan in the SIH paradigm are not paralleled by a blockade of stress-induced increases in plasma ACTH, corticosterone, and glucose levels. PMID- 8728566 TI - Effects of the novel cholecystokinin analogue Suc-Trp-N(Me)-Nle-Asp-Phe-NH2 on feeding and cortisol release in pigs. AB - Suc-Try-N-(Me)-Nle-Asp-Phe-NH2 is a succinylated tetrapeptide derived from the C terminal sequence of cholecystokinin octapeptide (CCK-B), which has been shown to have high agonist affinity for CCKB receptors. To test the validity of the hypothesis that implicates central CCKB receptors in the aetiology of stress related disorders, such as anxiety and panic, we argued that activation of these receptors by a CCKB receptor agonist should (i) suppress feeding motivation in hungry animals and (ii) increase plasma concentrations of the stress hormone cortisol. The effects of systemic and central administration of Suc-Try-N-(Me) Nle-Asp-Phe-NH2 were, therefore, investigated on operant food intake and cortisol secretion in pigs. Intravenous administration of Suc-Try-N-(Me)-Nle-Asp-Phe-NH2 (0.5-5 micrograms/kg) did not affect operant feeding in food-deprived pigs, although the highest dose (5 micrograms/kg) produced a small but significant (p < 0.05) increase in plasma cortisol levels 5-30 min after injection. Intracerebroventricular injection of Suc-Try-N-(Me)-Nle-Asp- Phe-NH2 (1-5 micrograms) had no effect on operant feeding and cortisol secretion in this species. The results obtained in this study indicate that central CCKB receptors are unlikely to be involved in stress-related behaviours in pigs. PMID- 8728567 TI - Mineralocorticoid and glucocorticoid receptor antagonists in animal models of anxiety. AB - The behavioral effects of intracerebroventricular (i.c.v.) administration of a specific mineralocorticoid receptor (MR) antagonist [RU28318 (10-50 ng/2 microliters)], a glucocorticoid receptor (GR) antagonist [RU38486 (1-50 ng/2 microliters)], or both antagonists (50 ng/2 microliters), were studied in two different animal models of fear and anxiety in rats. In the defensive burying paradigm simultaneous blockade of MR and GR increased immobility behavior, whereas a small decrease in defensive burying was seen. In the fear-potentiated startle test concurrent MR and GR blockade led to an increase in fear-potentiated startle at the highest loudness level (105 dB). In both tests the antagonists were not effective when given separately. The findings are discussed in terms of the involvement of GR and MR in neural mechanisms of fear and anxiety. PMID- 8728568 TI - Differential effects of gonadectomy on the thymocyte phenotypic profile in male and female rats. AB - As an organ responsible for generation of T-cell repertoire the thymus occupies a central position in establishment of mature immune response. To assess the potential role of the gonadal steroids in development and maintenance of immunological sexual dimorphism, the effects of gonadectomy pre- and postpuberty on the thymocyte profile of male and female rats were examined. Rats aged 30 days or 75 days were gonadectomized; 30 days later the thymic cellularity was estimated and the expression of the cell surface antigens (CD4 and CD8) and the T cell receptor (TCR) alpha beta was analyzed by flow cytometry. Regardless of age at surgery, the thymus weight and total thymocyte yield were greater in sham operated males than females; this sexual dimorphism in thymic cellularity persisted after gonadectomy. Sexual dimorphism in the composition of thymocyte subsets was also evident in sham-operated rats, with males expressing a higher percentage of CD4-8- cells, and remained after gonadectomy of adult rats. In male rats, gonadectomy at day 75 increased the percentage of CD4+8- single-positive and TCR alpha beta + cells. In contrast, in females, ovariectomy decreased the percentages of CD4+8- single-positive, CD4-CD8- double-negative, and TCR alpha beta + cells and increased the percentage of CD4+CD8+ double-positive cells. In the immature rats gonadectomy increased the percentages of CD4+CD8- single positive and TCR alpha beta + thymocytes and decreased the percentages of double positive and double-negative cells in males, while in the female it increased the percentage of CD4+8- single-positive thymocytes. Gonadectomy at that age abolished the sexual dimorphism in the expression of accessory molecules (i.e., CD4/CD8), but facilitated gender-specific expression of TCR alpha beta. In conclusion, the results suggest that the gonadal steroids are more important for the development than for the maintenance of the sexual dimorphism in the thymocyte composition. PMID- 8728569 TI - Met-enkephalin modulates stress-induced alterations of the immune response in mice. AB - Overnight restraint stress of mice decreased ConA-driven lymphocyte proliferation, plaque-forming cell response to sheep red blood cells (SRBC), and NK activity in the spleen, but the phagocytic activity was enhanced. Injection of methionine-enkephalin (MENK), 10 mg/kg, i.p., 30 min before restraint, abolished these changes (except for the NK activity) and attenuated the stress-induced elevation of glucocorticoids. However, MENK itself affected the immune responses like stress: It decreased NK activity and the PFC response and enhanced phagocytic activity. Contrary to results with stress, MENK had no effect on cell proliferation. The opioid-receptor antagonist naloxone given before restraint reversed the stress-induced enhancement of phagocytosis and the decrease of T cell proliferation. Alterations of the immune responses induced by restraint stress seem to be mediated by at least two mechanisms: activation of the hypothalamus-pituitary-adrenal (HPA) axis and the secretion of opioid peptides. MENK injected before stress may interfere with either or both mechanisms. T or B lymphocytes seem to be affected by the activation of the HPA axis, and phagocytes by a direct opioid action, whereas NK cells seem to be under the influence of another control mechanism. PMID- 8728571 TI - Stress and the immune system in the etiology of anxiety and depression. AB - There is clinical and experimental evidence that various aspects of the immune and endocrine systems are severely compromised in chronic stress and depression. For example, it has been shown that a reduced lymphocyte response occurs to mitogens in depressed patients, effects that are not reversed by chronic antidepressant treatment. By contrast, monocyte phagocytosis is increased, while neutrophil phagocytosis is decreased in depressed patients. Such changes are normalized by effective antidepressant treatment. The results of such studies and others that demonstrate alterations in noncellular immune processed in depression indicate that the changes in immune function correlate with the severity and duration of the external and/or internal stressful stimuli. There is evidence that some of the immune changes are a reflection of increased plasma glucocorticoids that characterize both stress and depression. However, it is also apparent that the cytokines, prostaglandins, and corticotrophic releasing factor (CRF) also play an important role in initiating the behavioral and pathophysiological changes that are characteristic of both depression and chronic stress. This review attempts to critically assess the interplay between CRF, the immune and neurotransmitter systems, and behavior in chronic stress and depression. PMID- 8728570 TI - Activation of the HPA axis by immune insults: roles and interactions of cytokines, eicosanoids, glucocorticoids. AB - It is now well established that challenges to the immune system (e.g., infection, inflammation) initiate diverse changes in neuroendocrine function, the most overt of which is activation of the hypothalamo-pituitary-adrenocortical (HPA) axis. The glucocorticoids that are released as a consequence fulfill a vital role in the maintenance of homeostasis that is effected in part through their ability to quench the immune/inflammatory response and thereby prevent them accelerating to a point where they become hazardous to the host. This article discusses the putative mechanisms by which immune insults stimulate the HPA axis, with particular reference to the roles and interactions of the interleukins, eicosanoids and glucocorticoids. PMID- 8728572 TI - Corticotropin releasing factor and its binding protein. AB - Although the lack of ACTH releasing activity of the high peripheral plasma levels of corticotropin releasing factor (CRF) of human placental origin can now be accounted for by the action of a specific sequestering plasma binding protein (pBP), there are many regions of the brain where the BP is found with little or no overlap with CRF. The existence of a mechanism promoting the rapid disappearance of pBP following bolus injection of exogenous CRF into normal individuals, which is triggered by the formation of a dimer complex (BP2/CRF2), and the elevation of pBP levels found in inflammatory disease, coupled with the lack of unequivocal evidence for endogenous CRF in many of these situations, suggests a role for pBP interaction with ligands other than CRF. We have searched for novel BP ligands in the brain and periphery and have found evidence for them in extracts of sheep brain and in synovial fluid collected from the joints of arthritic patients. These novel BP ligands could, thus, be the peptides responsible for many of the roles currently assigned to brain, peripheral, or immune CRF. PMID- 8728573 TI - Bacterial growth in saline implants: in vitro and in vivo studies. AB - The survival of bacteria was evaluated in custom-made saline breast implants with integral injection ports in vitro and in 10 New Zealand White rabbits for Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa, and Serratia marcescens. Pseudomonas and Serratia survived in vitro in saline-filled implants and multiplied 24-fold and 22-fold, respectively, from the initial inocula of 300 colony-forming units per cubic centimeter in 21 days. Serratia alone survived in saline implants placed on the dorsum of rabbits, proliferated 80-fold in 7 days, and tapered to 10-fold at the end of 3 weeks. Chemical analysis revealed the presence of glucose in fluid from the implants in the animal study (mean, 1.2 mg per deciliter; standard error of mean [SEM], 0.6) after 21 days and from human subjects (mean, 3.8 mg per deciliter; SEM, 1.0) after 8 months to 10 years. Serratia incubated in human breast implant fluid samples proliferated 7-fold to 30-fold greater than in the saline control in a nonaerated environment. We conclude that some bacteria are able to proliferate in saline in breast implants. Furthermore, their survival may be facilitated by a substance (i.e., glucose) that diffuses across the implant outer shell. PMID- 8728574 TI - Do saline breast implants harbor microbes? AB - Recent anecdotal reports have indicated that a high percentage of saline implants harbor dangerous microbes. These reports have caused considerable alarm and fear to saline implant recipients. Reports in the literature of microbial growth in saline implants are infrequent. The expander implant offers an ideal opportunity to study the internal milieu of saline implants in vivo. The study shows that, if used properly, saline implants do not become infected. In this study, 45 expander implants were studied under clinical conditions. No evidence of microbial contamination of the saline was found. PMID- 8728575 TI - Correction of severe blepharoptosis. AB - An analysis is made of 81 patients with severe blepharoptosis who underwent correction with autogenous fascia. In case of severe blepharoptosis, which means a levator function of less than 5 mm, correction by levator resection gives insufficient results. Better results are obtained by frontalis suspension, whereby the upper eyelid is connected to the frontalis muscle using fascia lata strips according to Crawford. The operative technique is described in detail. Particular interest has been paid to the width of the vertical lid fissure and symmetry between both upper eyelids after operation. The overall results were rated satisfactory to excellent with an average of 9 mm of postoperative vertical lid fissure. Comparison of the results of unilateral versus bilateral blepharoptosis correction revealed better results in the group of bilateral ptosis correction. In 76% of the bilateral patients, there was an asymmetry of less than 0.5 mm between both palpebral fissures, while in the unilateral group this result was achieved in only 35% of the cases. Symmetry is difficult to achieve in the unilateral group, especially when the vertical lid fissure of the nonptotic eye is 10 mm or more. In these cases, correction of both upper eyelids should be considered. The best time to operate on patients with severe congenital blepharoptosis is around the age of 4 to 5 years, because lagophthalmus after operation is better tolerated and the leg has developed sufficiently to provide an adequate amount of fascia. The correction of severe blepharoptosis by frontalis suspension with autogenous fascia shows a high rate of success and few complications. PMID- 8728576 TI - Rhomboid design for tubed inguinal flap in fingertip reconstruction. AB - The authors present a rhomboid design for the tubed inguinal flap mainly nourished by the superficial circumflex iliac vessels and the superficial inferior epigastric vessels. The length and width of the flap are mathematically determined based on the perimeter and length of the fingertip soft-tissue defect, respectively. The rhomboid tubed inguinal flap was used in the treatment of 21 patients with a variety of fingertip injuries. The thumb was involved in 12 patients and other digits in 9 patients. A complication, flap necrosis, occurred in one obese female patient and osteomyelitis of an amputated phalangeal bone developed in another. In all other patients, the rhomboid tubed inguinal flap healed uneventfully and provided satisfying results with minimal donor site morbidity. As a secondary procedure in thumb injuries, four neurovascular island flaps and one wrap around flap from the great toe were done for improved sensation. The precisely tailored rhomboid flap, with a safe and abundant vascular pedicle, allows for effective reconstruction of the fingertip with easy primary closure and minimal morbidity of the donor site. PMID- 8728577 TI - The role of vascular pedicle thrombectomy in the management of compromised free tissue transfers. AB - Microvascular free tissue transfer has become a very reliable reconstructive technique. Occasionally, flap compromise will occur and will require urgent flap reexploration. In the setting of complete thrombosis of pedicle vessels, thrombectomy using Fogarty #2 and #3 catheters can be effective in restoring vascular patency. Seven flaps with arterial, venous, or both vessel thrombosis were managed with thrombectomy at the time of reexploration and anastomotic revision. Six of the flaps were completely salvaged and the seventh was partially salvaged. Promptness in reexploration increased the likelihood of complete flap salvage. The use of postthrombectomy heparinization was associated with a 50% complication rat in this series. Thrombectomy of free flap vessels can be safely performed and is associated with a low incidence of rethrombosis. PMID- 8728578 TI - Effects of deferoxamine on ischemia/reperfusion injury after peripheral nerve compression. AB - We have demonstrated previously that acute nerve compression produces ischemia/reperfusion injury in rat sciatic nerve. In this study, we evaluated the effects of deferoxamine, an antioxidant, on recovery from ischemia/reperfusion injury after nerve compression. The sciatic nerves of male Sprague-Dawley rats, 370 to 430 g, were subjected to 24 hours of compression with Silastic tubing. The control group received intravenous saline solution at the time of decompression. The therapeutic group received intravenous deferoxamine (50 mg per kilogram) at the time of removal of the Silastic tubing. Nerve tissues within and distal to the compression site were assayed for malondialdehyde (MDA) levels and for growth associated protein 43 (GAP-43) expression, as markers of ischemia/reperfusion injury and nerve regeneration, respectively. In the control group (injury alone), the MDA levels were three times higher than normal during the initial 10 days and returned to normal by 14 days. In contrast, the deferoxamine treatment group had MDA levels that were not significantly different from precompression levels. In the control group, enhanced GAP-43 expression persisted until late in the recovery period. In the deferoxamine treatment group, the increased GAP-43 expression subsided early. The results suggest that the treatment of compressed peripheral nerve with deferoxamine at the time of surgical decompression reduces ischemia/reperfusion injury. PMID- 8728579 TI - The viability of cryopreserved onlay cranial bone allografts: a comparative experimental study versus fresh autografts. AB - It is well known that calvarial bone autografts are the bone grafts that are the least reabsorbent and have the best long-term evolution in craniofacial surgery. However, they do have certain limitations: (1) reabsorption results in repeated surgery and the need for new donor areas, (2) a limited amount of autogenous cranial bone is available (due to avoiding areas close to cranial sutures and venous sinuses, and because the temporal bone is very fragile and the cranium has not fully developed in children), and (3) graft extraction increases surgical time and morbidity. Because of this, we present an alternative to calvarial bone autografts: cryopreserved allografts. This paper is an experimental prospective study carried out on sheep with the following goals: (1) to assess the behavior of calvarial onlay bone grafts cryopreserved at -80 degrees C, using fresh autografts implanted under the same conditions as controls; (2) to compare reabsorption percentages statistically over time; and (3) to study qualitatively any histological variations. The results obtained are (1) more reabsorption of allografts when compared to autografts (at 90 days, 21.97% versus 20.21% of grafted volume), although this difference is not statistically significant; (2) a reduction in height in all onlay grafts as a consequence of the loss of the diploe; (3) the absence of any type of inflammation caused by a reaction to cryopreserved allografts; and (4) bone substitution performed using frozen allografts is histologically similar to that using fresh autografts. PMID- 8728580 TI - Assessing third-year medical students' knowledge of and exposure to cleft palate before and after plastic surgery rotation. AB - The purpose of this study was to evaluate third-year medical students' knowledge of and exposure to cleft palate before and after a rotation in plastic surgery. A 26-item questionnaire was administered to 37 students before and after their 12 week rotation that included the evaluation and management of patients with cleft palate. The questions addressed various aspects of the disorder, including identification of the members of a cleft palate team. Students were also asked about their clinical and academic exposure to cleft palate. When the students' responses to the 19 basic information items about cleft palate and associated problems were compared before and after the rotation, positive changes were documented for 14 of the 19 questions, minimal negative changes for 4, and no change for 1 question. The overall percentage of students indicating any type of clinical or academic exposure to cleft palate increased significantly after the rotation. Findings from this study suggest that an undergraduate clinical rotation in plastic surgery can increase the knowledge of and exposure to cleft palate. It was clear that supplemental learning can occur in other settings outside the classroom through interactions between faculty and students without additional curricular time. PMID- 8728581 TI - Rectosigmoid neocolpopoiesis for male-to-female transsexuals: Amsterdam experience. AB - Penile skin inversion is the method of choice for vaginoplasty in male-to-female transsexuals. Rectosigmoid neocolpopoiesis should be considered only when penile skin inversion has become impossible or has not led to functional results. In this paper we describe our technique and the results of colocolpopoiesis in 7 male-to-female transsexuals. PMID- 8728582 TI - Laser skin resurfacing. PMID- 8728583 TI - Q-switched ruby laser treatment of mucocutaneous melanosis associated with Peutz Jeghers syndrome. AB - Peutz-Jeghers syndrome is an eponym for circumscribed mucocutaneous melanosis in association with gastrointestinal polyposis. Irregular pigmented macules of varying size and color can be found on the perioral skin, lip vermillion border, buccal mucosa, palate, and tongue. Previous treatments, such as surgical excision, cryosurgery, electrodesiccation, dermabrasion, and carbon dioxide or argon laser ablation commonly result in incomplete removal, scarring, or changes in normal pigmentation. The Q-switched ruby laser used at 694 nm, a wavelength well absorbed by melanin relative to other optically absorbing structures in skin, causes highly selective destruction of pigment-laden cells. In addition, the 20-nanosecond pulse duration produced by this laser approximates the thermal relaxation time for melanosomes, thereby confining the energy to the target. The Q-switched ruby laser produces clinically significant fading of mucocutaneous melanosis in association with Peutz-Jeghers syndrome without complications often seen with other therapeutic modalities. PMID- 8728584 TI - Traumatic tattooing: treatment with the Q-switched ruby laser: a case study. AB - This case report details the successful treatment of extensive, facial traumatic tattooing in an adult male. The Q-switched ruby laser was utilized. It is demonstrated to be an excellent tool for the treatment of traumatic tattooing. The results achieved are superior to other methods and complications reported with other treatment options are minimized. PMID- 8728585 TI - A new technique of dermabrasion for traumatic tattoos. AB - We describe the use of a pinpoint bur to remove traumatic tattooing. The dermabrasion is concentrated on the affected tissues without damage to the surrounding normal skin. With this method, deeper areas of pigmentation are no more difficult to remove than superficial deposits. This technique is easily reproduced and highly effective. PMID- 8728586 TI - Use of modern craniofacial techniques for comprehensive reconstruction of the acromegalic face. AB - The severe acromegalic patient poses a difficult reconstructive dilemma to the craniofacial surgeon. Significant facial deformities can include frontal bossing, prominent supraorbital ridges, malar flatness, maxillary hypoplasia, mandibular prognathism with class III malocclusion, and macrogenia. Reports on the correction of these deformities are rare. Prior publications describe long hospital stays, weeks of intermaxillary fixation, requirement for a tracheostomy, as well as the need for multiple, staged procedures and interdisciplinary teams. In an effort to extend the advances of modern craniofacial techniques to this group of patients, we performed an extensive reconstruction on a 28-year-old acromegalic patient using a one-stage procedure without the use of intermaxillary fixation and without the added morbidity of a tracheostomy. The procedure addressed the skeletal deformities of the upper face, the midface, and the lower face. The operation was performed by a single plastic surgery team and the patient was extubated in 36 hours and discharged in 6 days. We believe that the use of rigid fixation and the judicious application of modern craniofacial principles can allow a complex yet safe one-stage procedure to reconstruct the acromegalic face. Such an approach showed decreased perioperative morbidity and provided an excellent functional and aesthetic result. PMID- 8728587 TI - A spheno-orbital encephalocele with unilateral exophthalmos. AB - Encephaloceles are rare congenital defects in the skull, through which the meninges and brain tissue have herilated. We report a case of an 11-year-old boy with a spheno-orbital encephalocele that presented with pulsatile exophthalmos. A two-stage operation resulted in a good outcome. PMID- 8728588 TI - Functional lower limb salvage with an osteocutaneous filet flap of the foot. AB - An osteocutaneous foot filet flap based on the posterior tibial vessels was successfully used to provide tibial coverage in a patient requiring a below-knee amputation following a high-voltage electrical injury. Addition of the calcaneus to the standard foot filet flap provided a vascularized bone graft that served to both lengthen the tibia and secure the flap via a tibial-calcaneal synostosis. The synostosis provided firm anchoring of the flap and allowed for a partial end bearing, below-knee prosthesis. PMID- 8728589 TI - Microsurgical reconstruction of difficult orocutaneous fistulas. AB - We present 5 cases from a series of eight orocutaneous fistulas that were successfully reconstructed through the use of free tissue transfer. Reconstruction was performed in 5 patients with a radial forearm flap and in 3 patients with a free jejunal transfer. This report addresses the approach to reconstructing the difficult orocutaneous fistula with emphasis on free flap design and the use of a bilaminar flap. PMID- 8728591 TI - Scrotal tourniquet injury due to thread ligation. PMID- 8728592 TI - Unilateral coronal synostosis (anterior plagiocephaly): current clinical perspectives. AB - Unilateral coronal synostosis is a frequently seen form of craniosynostosis that results in an anterior plagiocephalic shape to the ipsilateral forehead and orbit. Compensatory bulging of the contralateral side always occurs and most surgeons agree that a bilateral reconstruction is required. Details of the timing and techniques to achieve a symmetrical, proportional upper face will vary according to the craniofacial center's philosophy, but a team approach, with the pediatric craniofacial surgeon and the pediatric neurosurgeon working together, is an essential ingredient to achieving an unobstrusive face in a child born with unilateral coronal synostosis. More accurate methods of documenting the presenting deformity and the initial and late reconstructive results are needed. PMID- 8728590 TI - The "shutter flap" for large defects of the forehead. AB - Acquired defects involving exposed bone in the central forehead can be challenging to reconstruct. We present a simple technique in which two superiorly based flaps of galea frontalis and skin are transposed medially, like the shutter of a camera. Full-thickness grafts are used to close the donor site. This is a remarkably easy way to close some very large forehead wounds. PMID- 8728593 TI - A computer heart model incorporating anisotropic propagation. III. Simulation of ectopic beats. AB - With the advent of catheter ablation procedures, it has become an important goal to predict noninvasively the site of origin of ventricular tachycardia. Site classifications based on the observed body surface potential maps (BSPMs) during ventricular endocardial pacing, as well as on the patterns of the QRS integrals of these maps, have been suggested. The goals of this study were to verify these maps and their QRS integral patterns via simulation using a computer heart model with realistic geometry and to determine whether the model could improve clinical understanding of these ectopic patterns. Simulation was achieved by initiating excitation of the heart model at different endocardial sites and their overlying epicardial counterparts. This excitation propagated in anisotropic fashion in the myocardium. Retrograde excitation of the model's His-Purkinje conduction system was necessary to obtain realistic activation durations. Simulated BSPMs, computed by placing the heart model inside a numerical torso model, and their QRS integrals were close to those observed clinically. Small differences in QRS integral map patterns and in the positions of the QRS integral map extrema were noted for endocardial sites in the left septal and anteroseptal regions. The simulated BSPMs during early QRS for an endocardial site and its epicardial counterpart tended to be mirror images about the zero isopotential contour, exchanging positive and negative map regions. The simulation results attest to the model's ability to reproduce accurately clinically recorded body surface potential distributions obtained following endocardial stimulation. The QRS integral maps from endocardial sites in the left septal and anteroseptal regions were the most labile, owing to considerable cancellation effects. Conventional BSPMs can be useful to help distinguish between endocardial and epicardial ectopic sites. PMID- 8728594 TI - A computer heart model incorporating anisotropic propagation. IV. Simulation of regional myocardial ischemia. AB - The main goal of this study was to simulate clinical body surface potential maps, recorded during percutaneous transluminal coronary angioplasty protocols, using a realistic geometry computer heart model. Other objectives were to address the question of reciprocal ST-segment changes observed in the 12-lead electrocardiogram during ischemia and to verify the hypothesis that the shortening of the QRS duration observed in left anterior descending (LAD) coronary artery occlusion may be explained by conduction delay in the septal His Purkinje system. Simulation was achieved by first introducing into the heart model three transmural zones of mild, moderate, and severe ischemia for assumed occlusions in the LAD, left circumflex, and right coronary arteries. The heart model was then excited, in turn, with these three zones present for assumed occlusions in the LAD, left circumflex, and right coronary arteries. Myocardial conduction velocities in the regions of moderate and severe ischemia were assumed to be reduced to 75 and 50% of normal, respectively. Model action potentials in the mild, moderate, and severely ischemic zones were also altered to reflect known ischemic changes in these action potentials. Body surface potential maps and electrocardiograms were computed by placing the heart inside a numerical torso model. Simulated map patterns during both ST-segment and QRS were qualitatively similar to clinical maps. Reciprocal ST-segment depression was observed for all three occlusions in remote leads that did not overlie the ischemic zones. QRS shortening due to septal His-Purkinje conduction delay was verified. The simulation results attest to the model's ability to reproduce body surface potential distributions recorded following percutaneous transluminal coronary angioplasty protocols. The simulations also showed that reciprocal ST segment changes occur as a natural consequence of the primary ischemic region and that there is no need to invoke a second region of ischemia. Finally, the model demonstrated that QRS shortening can occur in LAD occlusion despite a slowing of conduction down the septal His-Purkinje system. PMID- 8728595 TI - Sex differences and frequency content of the Frank lead signal-averaged ECG in a normal population. AB - To quantify the content of the signal-averaged electrocardiogram (ECG) in normal subjects, 100-beat signal-averaged Frank lead ECGs at a sampling rate of 1,000 Hz and with 16-bit resolution were recorded from 88 women and 102 men with a combined average age of 22 years. The QRS portion of each lead was digitally filtered in four bandwidths: 0-10, 10-60, 60-150, and 150-250 Hz. The root-mean square voltage of each filtered signal was calculated as an absolute value and normalized as a percentage of the sum of the four filters. Women had significantly lower root-mean-square values in all filtered bands in Frank leads X and Z (P < .01). Normalized data, however, showed no sex difference (P > .05). Sex differences may affect quantitative measurement of the frequency content of the signal-averaged ECG. PMID- 8728596 TI - Usefulness of QRST isointegral maps for the diagnosis of right ventricular pressure overload in patients with surgically repaired tetralogy of Fallot complicated by right bundle block. AB - Right ventricular pressure overload was evaluated in 29 patients, 8-12 years old, with surgically repaired tetralogy of Fallot by using body surface QRST isointegral maps. In patients with right ventricular systolic pressure above 50 mmHg, the maxima of the isointegral maps tended to shift toward the lower right hand region of the map. The maximum value was significantly correlated with right ventricular systolic pressure (r = .58; P < .01). There was a correlation between the right ventricular systolic pressure and the percentage +2SD and percentage +5SD departure areas, which are defined as the area (expressed as a percentage of the total chest area) in which the QRST integral values are greater than the normal mean +2SD or +5SD, respectively (r = .61 and .84, P < .01). The QRST isointegral map can be used to evaluate right ventricular pressure overload in postoperative patients with tetralogy of Fallot complicated by right bundle branch block. The percentage +5SD departure area is the most valuable parameter for the quantitative evaluation of the right ventricular systolic pressure. PMID- 8728597 TI - Electrocardiographic diagnosis of infarction of the right ventricular anterior wall. AB - Fifty cases of acute myocardial infarction manifesting in anterior chest leads were studied. The site of infarction was determined by detailed two-dimensional echocardiography. Isolated left ventricular anteroseptal infarction was found in 32 patients, infarction of the anterior wall of the left as well as the right ventricle in 12, and infarction localized to the right ventricular anterior wall in 6. ST-segment depression followed by deep symmetrical T wave inversion, increasing in depth from lead V1 to lead V3 without loss of the R wave in these leads, was highly sensitive and specific in detecting isolated right ventricular anterior wall infarction. ST-segment elevation in right-sided chest leads was not useful. Concomitant ST-segment depression in inferior leads was highly specific and sensitive in diagnosing infarction of the right ventricular anterior wall in the presence of left ventricular anterior infarction. PMID- 8728599 TI - Reverse effects of exercise on the sinus and parasystolic cycle lengths. AB - Ventricular arrhythmias are the primary concern in the exercise laboratory. It has recently been suggested that in not a few cases, ventricular premature complexes are governed by modulated parasystole and not by ordinary extrasystolic rhythm. In many cases, however, it is difficult to differentiate between parasystole and ordinary extrasystoles. Few reports are available on the effect of exercise in cases of "true" parasystole. This study investigated the effect of exercise on the parasystolic cycle length 11 cases of true ventricular parasystole, in which one or more "pure" parasystolic cycles containing no intervening nonectopic QRS complexes were found. In all cases, in contrast to an acceleration of the sinus rate, a definite decrease in the parasystolic rate was found. In no case did complete suppression occur. These findings suggest that the effects of exercise on ordinary ventricular extrasystolic rhythm and on ventricular parasystole may be considerably different from each other. PMID- 8728598 TI - Mechanisms of the in vitro effects of amphetamine on rat sinus node automaticity and membrane potentials of atrial fibers. AB - The main objective of this investigation was to clarify the mechanisms of the acute in vitro actions of amphetamine (AMP) on cardiac electrophysiology. Concentrations of AMP ranging from those considered clinically therapeutic to those considered toxic were tested in isolated rat sinoatrial tissues while recording membrane potentials with intracellular microelectrodes. In preparations beating spontaneously, 6.8 nM-2.71 microM AMP exerted a positive chronotropic action that was blocked by propranolol. The positive chronotropic action of 5.43 microM AMP was smaller than that of 2.7 microM AMP and was reversed by propranolol. Neither phentolamine nor atropine blocked this depressant action of AMP. It is concluded that the positive chronotropic action of AMP was beta adrenergic and that beta-adrenergic block unmasked a negative chronotropic action of a high concentration of AMP, which was neither alpha-adrenergic nor muscarinic. In atrial fibers driven at a constant rate, 54.3 nM AMP prolonged the action potential duration (APD), without affecting the resting membrane potential (RMP), the action potential amplitude (APA), or the maximum velocity of phase 0, while 5.43 microM AMP reduced RMP, APA, and the maximum velocity of phase 0, and increased APD. The prolongation of APD, as well as the decreases of RMP and APA, was not abolished by propranolol, phentolamine, or 4-aminopyridine. Conversely, nifedipine abolished the effects of AMP on all three parameters. In general, AMP produced mainly a prolongation of the action potential. Only a high concentration of AMP decreased RMP and depressed phase 0 of the action potential. The effect of AMP on APD, RMP, and APA essentially involved increasing the influx of calcium through the L-type channels in the sarcolemma. PMID- 8728600 TI - Body surface potential mapping of a patient with Wolff-Parkinson-White syndrome with two accessory pathways and two atrial pacemaker complexes. AB - As part of an ongoing research protocol, a patient with Wolff-Parkinson-White syndrome underwent body surface potential mapping and electrophysiologic studies before radiofrequency ablation therapy. Careful analysis of the body surface potential mapping data made it possible to distinguish four different map sequences representing four different cardiac complexes. Analysis of these maps is consistent with two accessory pathways, with the additional pathology of two distinct atrial pacemaker sites. A right anterosuperior pathway was found to conduct continuously. The second pathway is consistent with a right inferior pathway conducting intermittently. The analysis demonstrates the type of information that can be extracted from body surface potential maps, even in the presence of complex pathologies. PMID- 8728601 TI - Nonreentrant supraventricular tachycardia due to double ventricular response via dual atrioventricular nodal pathways. AB - Narrow and wide QRS tachycardias associated with various rhythm disturbances were recognized during 24-hour ambulatory eletrocardiographic monitoring in a 65-year old man with coronary artery disease. Laddergram analysis revealed the presence of dual atrioventricular nodal pathways. Non-reentrant supraventricular tachycardia due to simultaneous fast and slow conduction through the dual atrioventricular nodal pathways was confirmed by electrophysiologic study. The atrial rate determined the occurrence of simultaneous conduction, and extrastimulation failed to induce a double ventricular response. Enhanced vagal activity was thought to play a critical role in provoking this phenomenon. Radiofrequency catheter ablation of the slow pathway eliminated the arrhythmias. PMID- 8728602 TI - Ventricular tachycardia with narrow QRS duration, a right bundle branch block pattern, and right axis deviation abolished by catheter manipulation. AB - A 25-year-old women underwent electrophysiologic evaluation for sustained normal QRS complex tachycardia with a pattern of right bundle branch block and right axis deviation. Ventricular tachycardia was diagnosed by demonstrating fusion beats, atrioventricular dissociation, and bundle of His potential activation, which began before the onset of each QRS complex. A single ventricular extrastimulus was capable of easily provoking the tachycardia. There was an inverse relationship between the coupling interval of the first extrastimulus and the interval of the first tachycardia beat, suggesting reentry as the mechanism. The tachycardia was unexpectedly abolished during catheter manipulation in the left ventricle and has never recurred during 1 year of follow-up evaluation. The tachycardia was thought to be an unusual form of interfascicular tachycardia or microreentrant fascicular tachycardia. PMID- 8728603 TI - Bacterial responses to host-defense peptides. PMID- 8728604 TI - Sleepless in Bologna: transmission of fatal familial insomnia. PMID- 8728605 TI - Role of Epstein-Barr virus gene latent membrane protein in nasopharyngeal carcinoma. PMID- 8728606 TI - Running the gamut of retroviral variation. AB - The retroviral DNA polymerase, or reverse transcriptase, lacks a 3' exonuclease proofreading activity. This causes a high mutation rate with the result that genetic diversity and drug resistance are increased. Only combination chemotherapy has the potential to out-manoeuvre these viruses. However, there is no evidence that ongoing variation allows escape from immune surveillance, so contributing to pathogenesis. PMID- 8728607 TI - Understanding signal transduction during bacterial infection. AB - Many known or suspected bacterial virulence factors require environmentally responsive control factors for expression. In Bordetella species, the BvgAS system represses and activates sets of genes, and mediates a biphasic phenotypic transition. Studies using mutants with altered signaling pathways and reversed regulatory connections have provided insights into the role of BvgAS and this phenotypic transition during the Bordetella-host interaction. PMID- 8728608 TI - Role of verotoxin receptors in pathogenesis. AB - Verotoxin-globotriaosyl ceramide (Gb3) binding is the linchpin in disease induced by verotoxin-producing Escherichia coli (VTEC), and defines cell sensitivity, tissue tropism, mode of systemic transport, specific cytotoxic activity and internal routing within sensitive cells. Binding explains the epidemiology of renal pathology, which may follow VTEC infection. Lipid heterogeneity of Gb3 is important in binding, and may define a growth-related signal transduction pathway used by verotoxin. PMID- 8728609 TI - Seed transmission of plant viruses: a lesson in biological complexity. AB - Seed transmission of plant viruses results from the three-way interplay of the genetic components of the virus, the maternal host and its progeny. The challenge is to use genetic and cell biological analyses, integrated with our knowledge of plant reproduction and embryo development, to dissect this complex and rather poorly understood process. PMID- 8728610 TI - Cohabitation of Leishmania amazonensis and Coxiella burnetii. AB - Intracellular pathogens customize the composition and function of the vacuoles they occupy, and can arrest or distort vacuolar maturation. In doubly infected cells, vacuoles that contain two different parasites can be used to test for exclusionary mechanisms, for expression of vacuolar phenotypes that permit or restrict fusion, and for the survival of pathogens targeted to an unusual cellular compartment. PMID- 8728611 TI - The prospects for developing a vaccine against Staphylococcus aureus. AB - Staphylococcus aureus is an important bacterial pathogen with multiple virulence factors. The prevalence of antibiotic resistance among clinical isolates means that new vaccine strategies to prevent staphylococcal infections are needed. An ideal vaccine would induce antibodies to prevent bacterial adherence, promote opsonophagocytic killing by leukocytes and neutralize toxic secreted proteins. PMID- 8728612 TI - A question of sunglasses. PMID- 8728613 TI - Oerskovia xanthineolytica keratitis. PMID- 8728614 TI - Clinical study of disposable contact lens wear: Brazilian experience. AB - PURPOSE: The objective of this study was to analyze prospectively the continuous use of disposable contact lenses. METHODS: We conducted a clinical study of extended wear of disposable contact lenses for a period of 7 days and 6 nights by 25 patients during a 90-day period. Factors such as visual acuity, keratometry, complications, and contamination were analyzed. RESULTS: Visual acuity and keratometry measurements did not present significant alterations during the study. Complications related to the use of disposable contact lenses were not observed. Analysis of cultures did not present any significant microbiological alterations due to contact lens use. All of the patients were satisfied with the wear schedule and considered the Acuvue lenses comfortable. CONCLUSIONS: Our results may be useful in increasing the level of acceptance of extended wear of disposable contact lenses in Brazil. PMID- 8728615 TI - Indications and complications of therapeutic disposable Acuvue contact lenses. AB - PURPOSE: Although therapeutic hydrogel contact lenses have been used for many years, there have been few reports on the therapeutic use of disposable lenses. We evaluated lens fit, duration of wear, efficacy of therapy, complications, and concurrent treatment in patients with corneal or anterior segment problems who were managed with Acuvue contact lenses for therapeutic purposes. METHODS: We managed 38 patients with corneal or anterior segment disease during 1992-1994. The duration of lens wear ranged from several days to 12 months. Six patients were fit for mechanical reasons, six were fit for symptomatic relief, and 26 were fit as an adjunct to healing. Technical failures included lenses with inadequate movement and lenses that fell out of the eye. Therapeutic failures occurred when lens use did not help resolve the anterior segment problem. RESULTS: Seventy-one percent of patients were fit successfully, and 66% of patients were therapeutic successes. Complications with contact lens fitting included lens loss, discomfort, corneal ulceration, and tight lens syndrome. CONCLUSIONS: Although we found some difficulty in lens fitting, Acuvue lenses may provide a useful, disposable, and less expensive alternative to the traditional bandage contact lens. PMID- 8728616 TI - A quantitative and qualitative assessment of the NOVAWET-Perception bifocal contact lens. AB - PURPOSE: We prospectively evaluated the NOVAWET-Perception multifocal contact lens for comfort and performance. METHODS: We fit 28 presbyopic patients with this multifocal contact lens. This multifocal rigid gas permeable lens has a multiaspheric back surface and multiple eccentricity values, allowing for a natural field of vision at all angles. Patients were evaluated prior to inclusion in the study and then at 1,3, and 9 months. Evaluations included slit lamp examination, visual acuity, contrast sensitivity, and completion of a questionnaire regarding comfort and overall satisfaction. RESULTS: At the end of the study, 14 of 28 patients (50%) were successfully wearing the NOVAWET Perception lenses. Of these, 12 (86%) achieved J2 or better near vision and 13 (93%) achieved 20/25 or better distance vision. Fourteen patients failed to complete the study. CONCLUSIONS: We believe the NOVALENS multifocal contact lens is a viable option for the correction of many presbyopic patients. PMID- 8728617 TI - The use of synthetic Cecropin (D5C) in disinfecting contact lens solutions. AB - PURPOSE: Microbial keratitis caused by Pseudomonas aeruginosa is the most common contact lens associated corneal infection. Cecropins are microbicidal peptides isolated from the hemolymph of the Cecropia moth. Previous in vitro studies have demonstrated their efficacy against a broad spectrum of ocular pathogens. This study was designed: a) to evaluate the antimicrobial potency of three different contact lens solutions (Renu, Complete, and Opti-Free) against P. aeruginosa, and b) to evaluate the activity of the same contact lens solutions in combination with a synthetic cecropin analog, D5C, against the challenge organism in the presence of a soft contact lens. METHODS: A virulent strain of P.aeruginosa isolated from a case of ulcerative keratitis was used in the study. Three different concentrations of bacteria (10(3), 10(5) and 10(7) CFU/mL) were inoculated into the contact lens solutions and into buffered saline, which was employed as a control. The samples were incubated at 27 degrees C, and at time 0, 30, and 90 minutes, 24, 48, and 72 hours, and aliquots of the test solutions were plated and subcultured on nutrient agar. After 24 hours of incubation at 37 degrees C, colonies observed on the nutrient agar plates were counted. To study the antimicrobial efficacy of D5C (100 micrograms/mL), we used the identical test series and assay, adding a soft contact lens to the solutions and a larger inoculum of bacteria (10(9) CFU/mL). RESULTS: After 72 hours, all of the contact lens solutions tested sterilized 10(3) CFU/mL of P. aeruginosa. At 10(7) CFU/mL, they yielded greater than 2 logs of killing of the bacteria, but the solutions were not sterilized. The addition of D5C (100 micrograms/mL) to the contact lens solutions yielded greater than 3 logs of killing with a larger inoculum of bacteria in the presence of the soft contact lens. CONCLUSION: The contact lens solutions tested were effective against P. aeruginosa at 27 degrees C for up to 72 hours with an inoculum of 10(3) CFU/mL. The addition of D5C augmented their antimicrobial activity in the presence of the contact lens. PMID- 8728618 TI - Comparative disinfectant efficacy of two disinfecting solutions against Pseudomonas aeruginosa. AB - PURPOSE: We compared the disinfecting efficacy of Pure Eyes Disinfectant and Opti Free Rinsing, Disinfecting and Storage Solution, without recommended daily cleaning steps, against Pseudomonas aeruginosa. METHODS: Vifilcon A soft contact lenses were inoculated with the challenge organism, P. aeruginosa ATCC 15442, at a concentration of 1 x 10(8) colony forming units (CFU/mL) and disinfected with each solution for four and six hours, respectively. RESULTS: Pure Eyes eradicated P. aeruginosa after both four and six hour soak times. With all lots of Opti Free, colony forming units were too numerous to count (> 200) for both soak times. CONCLUSIONS: We concluded that, because Pure Eyes disinfecting solutions can eradicate P. aeruginosa without the benefit of a daily cleaning step, this disinfecting process may offer protective benefits to noncompliant contact lens wearers. PMID- 8728619 TI - Non-contact specular microscopic observation for early response of corneal endothelium after contact lens wear. AB - PURPOSE: Contact lenses induce short- and long-term corneal endothelial changes, including endothelial blebs. The endothelial bleb is a reversible, short-term response found on the corneal endothelium after lens wear. We used non-contact specular microscopy to observe endothelial blebs occurring with different types of contact lenses. METHODS: We observed the time course, frequency, and location of endothelial blebs in 11 eyes of 9 contact lens wearing patients. Eight types of contact lenses with various oxygen transmissibilities (Dk/L) were used in the study. RESULTS: Blebs were observed in the central cornea in patients with polymethylmethacrylate and rigid gas permeable contact lenses, while blebs in the central to mid-peripheral cornea were seen in patients wearing soft contact lenses. CONCLUSIONS: Although the number of blebs vary by individual patient, there appears to be an inverse correlation between the number of blebs and the Dk/L of the contact lens. PMID- 8728620 TI - Transient corneal stromal and endothelial changes following soft contact lens wear: a study with confocal microscopy. AB - PURPOSE: Previous studies have described transient corneal endothelial changes in non-contact lens wearers after a short period of soft contact lens wear by means of contact and noncontact specular microscopy and modified slit lamp biomicroscopy, which provide magnifications from 60 to 100x. In this investigation, we documented and characterized these contact lens-related corneal changes using the white light, real-time confocal microscope, which is capable of cellular resolution imaging of all layers within the cornea at magnifications of 100 to 500x. METHODS: We used a clinical confocal microscope to study corneal changes in three patients wearing a high water content soft contact lens for the first time. RESULTS: In one patient, endothelial changes consisting of irregularly shaped, round or oval, dark regions were observed within the endothelial mosaic. Scattered hyper-reflective keratocyte nuclei were seen in the posterior stroma. The keratocytic and endothelial changes were most evident 20 minutes after placement of the lens. By 30 minutes, the changes were fewer and less prominent, and the brightness of the highly reflective keratocyte nuclei had decreased. CONCLUSIONS: These studies show, for the first time, that the transient changes associated with contact lens wear occur not only in the endothelium, but also in the corneal stroma. It has been suggested that the changes result from an increase in CO2 and lactic acid, which causes a transient reduction in the corneal pH. We hypothesize that the resulting acidic environment may induce gene expression that causes changes in the involved nuclei, which in the keratocytes become hyper-reflective, and in the endothelium become enlarged, resulting in posterior displacement of the cell membrane and producing the dark "blebs" and irregular lines observed at this level of the posterior cornea. PMID- 8728621 TI - Changes in corneal radius and thickness in response to extended wear of rigid gas permeable contact lenses. AB - PURPOSE: In this study we assessed patients' clinical tolerance to extended wear of Hoya Hard EX contact lenses, which have a Dk of 125, and determined resulting changes in corneal radius and thickness. METHODS: Hoya Hard EX rigid gas permeable contact lenses (Dk = 125) were fitted to 57 eyes of 29 patients; all eyes were free of pathology. The average duration of contact lens wear was 5.28 +/- 1.43 days. Changes in the radius and thickness of the cornea were measured after 1,3, and 6 months of lens wear. RESULTS: At 1 month, keratometer readings indicated significant flattening in both the horizontal (P < 0.01) and vertical meridians (P < 0.001). At 3 and 6 months, the flattening of both meridians had become more significant (P < 0.001). Although there were no statistically significant changes in corneal thickness at 1 month, significant decreases in thickness were observed at 3 and 6 months (P < 0.001). During the study, patients' vision was stable and lens wear caused no serious complications. At the end of the study, all 29 patients continued extended wear of their contact lenses. CONCLUSIONS: High-Dk RGP contact lenses are an alternative to soft lenses for extended wear. PMID- 8728622 TI - Initial treatment of microbial keratitis. AB - PURPOSE: The common occurrence of failed medical treatment in microbial keratitis led us to investigate this phenomenon. METHODS: We retrospectively reviewed all ulcers that presented to our department for 24 consecutive months. We classified each ulcer as either a therapeutic success or failure based on a precise definition of the response to initial antibiotic selection. We then analyzed multiple factors including: antibiotic selection, ophthalmic disease, ulcer characteristics, and management, to determine their significance in the success or failure in treating microbial keratitis. Complications were also examined. RESULTS: Important factors in failure were non-fortified antibiotics (P < 0.001), ocular surface disease (P = 0.0178) and outpatient management (P < 0.001). Large ulcers (P = 0.051) were of borderline significance. Sensitivity results reflect high sensitivity among successfully treated patients when appropriate antibiotics are chosen. CONCLUSIONS: This report provides insight into current practice patterns and potential means to improve success in managing microbial keratitis. PMID- 8728623 TI - Dry eye: pharmacological approaches, effects, and progress. AB - PURPOSE: The effects of available pharmaceuticals on the tear film and ocular surface in dry eye are reviewed and the progress that has been made in dry eye treatment is discussed. METHODS: Pharmaceuticals are examined with regard to their effect on physical and morphological events or milestones in the natural history of dry eye disease. RESULTS: Dry eye disease evolves cumulatively over time through a sequence of four events: 1) decreased tear production or increased tear film evaporation (increased tear osmolarity); 2) decreased conjunctival goblet-cell density; 3) increased corneal epithelial desquamation; and, finally, 4) destabilization of the cornea-tear interface. In the 1970s, demulcents were introduced to decrease tear film instability. More recently, preservative-free solutions have been shown to decrease corneal desquamation. The newest pharmaceutical available today is known as TheraTears (ATF); in pre-clinical studies it restores conjunctival goblet cells. On the horizon are medications such as 3-isobutyl-1-methylxanthine (IBMX), which directly stimulate tear production. CONCLUSIONS: As our understanding of the pathogenesis and natural history of dry eye disease has improved, pharmaceuticals have been designed to act at earlier and more crucial points in the disease process. We are now entering a new era in dry eye treatment with the introduction of the first agent that can reverse the basic events of dry eye surface disease. PMID- 8728624 TI - Corneal melting in a chronic alcoholic contact lens wearer. AB - PURPOSE: We report a case of bilateral keratitis in a 43-year-old alcoholic patient who wore soft daily wear contact lenses. METHODS: After consecutive negative cultures of corneal and conjunctival scrapings and after observing no improvement following antibiotic therapy, the patient was clinically diagnosed with vitamin A deficiency. RESULTS: After 20 days of treatment with vitamin A, the patient's cornea completely healed. Laboratory results confirmed the clinical diagnosis. CONCLUSIONS: When the ophthalmologist is confronted with corneal melting in a known alcoholic or drug-user, it is imperative to consider the possibility of vitamin A deficiency as a possible cause, especially in the contact lens wearer. PMID- 8728625 TI - Irreversible bullous keratopathy after air bag trauma. AB - PURPOSE: We describe an unusual case of corneal decompensation following air bag trauma in a 38 year-old-female. METHODS: The patient was followed clinically for 5 months post-injury with persistent corneal edema that failed to resolve, necessitating corneal transplantation. Scanning electron microscopy (SEM) of the corneal button was performed. RESULTS: SEM revealed localized areas of complete endothelial destruction. Other areas of the endothelium had cell counts under 1,000 cells per square millimeter. CONCLUSIONS: This case corroborates animal studies demonstrating endothelial cell loss from direct air bag trauma and stresses the need to possibly redesign this safety device for maximum driver protection. PMID- 8728626 TI - Neuromuscular scoliosis: recent concepts. PMID- 8728627 TI - Reconstructive spine surgery in pediatric patients with major loss in vital capacity. AB - Thirty-two pediatric patients with severe restrictive lung disease identified with vital capacities < 40% of predicted, who had undergone major reconstructive spine surgery, were reviewed. There were 18 boys and 14 girls, the mean age was 13 years (range, 7-17), and the mean vital capacity was 31% of predicted (range, 16-39%). Fifty-four procedures were performed, 13 posterior only, one of which was staged, and 19 anterior and posterior procedures, of which 15 were staged and four were sequential. The incidence of pulmonary complications (pneumonia, reintubation, pneumothorax, respiratory arrest, or the need for tracheostomy) was 19% (six patients), and only three patients required tracheostomy. The surgical and perioperative mortality rate was zero. Patients who had a thoracotomy or a thoracoabdominal approach had a significantly higher number of pulmonary complications. The use of preoperative decreased vital capacity as a measure of inoperability excludes the young patient most in need of surgical intervention. With improved preoperative, intraoperative, and postoperative techniques, careful monitoring, and the cooperation of pediatric pulmonologists and intensivists, reconstructive spine surgery can be performed in the pediatric patient with severe decreased vital capacity with very acceptable morbidity and mortality. PMID- 8728628 TI - Same-day versus staged anterior-posterior spinal surgery in a neuromuscular scoliosis population: the evaluation of medical complications. AB - The medical complications occurring in 29 patients with neuromuscular spinal deformity undergoing two-stage anterior-posterior spinal fusion ("staged") were compared with 16 neuromuscular patients undergoing single-stage anterior posterior spinal fusion ("same day"). Thirty-six (124%) major and minor medical complications occurred postoperatively in the staged patients, whereas 14 (88%) major or minor complication were present in same-day surgery patients. Thirty five percent of staged patients had no complications, whereas 63% of same-day patients were without complications. Associated findings comparing the two-stage procedure to the single-stage surgery included operative and anesthesia time increase, increased blood-volume loss, increased blood transfusion, decreased nutritional parameters, and longer hospital stays. With either approach, there is the risk of significant complications in this vulnerable population. PMID- 8728629 TI - Posterior instrumentation and fusion of the thoracolumbar spine for treatment of neuromuscular scoliosis. AB - We reviewed the clinical and technical outcomes of 25 patients with neuromuscular scoliosis, who were treated by Luque instrumentation and posterior spinal fusion from the upper thoracic spine to L5 between 1981 and 1988. A mean curve correction of 46% was obtained operatively with a mean 8 degrees loss of correction during the follow-up period that ranged from 1.9 to 9.4 years (mean, 5.5). Pelvic obliquity was improved 50% from a mean of 16.1 degrees to a mean of 8.1 degrees in 24 patients for whom data were available. At final follow-up, the mean pelvic obliquity increased to 11.4 degrees with only two patients increasing > 8 degrees. The cause for major postoperative increase in pelvic obliquity was continued anterior spinal growth with torsion of the fusion mass and was not related to changes limited to the L5-S1 motion segment. Posterior fusion and instrumentation from the upper thoracic spine to L5 without anterior fusion provides adequate correction and control of spinal deformity for many patients with cerebral palsy. Those patients with significant growth remaining, or with severe deformities, may benefit by preliminary anterior release and fusion or inclusion of the pelvis and sacrum. PMID- 8728630 TI - Posterior spinal fusion for scoliosis in patients with cerebral palsy: a comparison of Luque rod and Unit Rod instrumentation. AB - The development of the U-shaped Unit Rod for posterior spinal arthrodesis is a recent advance in the treatment of spinal deformity in patients with cerebral palsy. The results of 15 patients who underwent arthrodesis with dual Luque rod instrumentation (group I) are compared with the results of 15 patients in whom Unit Rod instrumentation was used (group II). The two treatment groups were similar with respect to age, gender, major spinal curve, and degree of pelvic obliquity. The Unit Rod instrumentation allowed significantly greater correction of both the major curve and pelvic obliquity, as assessed on postoperative radiographs. The mean postoperative major curve in group I was 44.1 degrees, compared with 31.7 degrees in group II (mean corrections of 48.6 and 61.7%, respectively). The mean angle of postoperative pelvic obliquity in group I was 12.6 degrees, compared with 5.2 degrees in group II, corrections of 49.5 and 79.3%, respectively. Sagittal-plane alignment was improved to a similar degree in both groups in those patients with preoperative imbalance and maintained in the remainder of patients. PMID- 8728631 TI - Spinal fusion in Duchenne's muscular dystrophy. AB - The Women's and Children's Hospital experience with Luque spinal fusion in Duchenne's muscular dystrophy was reviewed from its commencement in 1983 to the present with a view to assessing the clinical and radiologic outcome and safety of the procedure. Seventeen boys have undergone spinal fusion. L-rod instrumentation was used in 10, six of whom had significant problems with sitting imbalance or progression of the scoliosis or both. In seven cases, distal instrumentation was taken to the pelvis with a Galveston construct and rigid crosslinking. Apart from some progression and sitting imbalance in the L-rod group, there were few complications. In the Galveston group, pelvic obliquity was corrected by a mean of 63%, and there was better maintenance of correction. There were no pseudoarthroses or instrument failures in the Galveston group. Of the total group, four patients had forced vital capacity (FVC) values < 25% predicted, and two required ventilation postoperative (< 48 h). There were no other respiratory complications. The effect of surgery on respiratory function remains uncertain. Spinal fusion with the Luque rod construct and pelvic fixation is a safe procedure. It provided a mean correction of 60% and control of pelvic obliquity without significant postoperative deterioration. In our experience, surgery can be safely performed with FVC value down to 20% predicted. On the basis of these data, one current practice is to instrument to the pelvis with a Galveston construct and Texas Scottish Rite Hospital cross-linking. PMID- 8728632 TI - Impact of orthoses on the rate of scoliosis progression in children with cerebral palsy. AB - This study was undertaken to determine the impact of spinal bracing on curve pattern and the rate of progression of neuromuscular scoliosis in children with cerebral palsy. Twenty-one patients were treated with a Wilmington custom-molded orthosis with 23 h a day brace wear for a mean bracing period of 67 months (range, 22-173). Twenty-two patients had a similar follow-up to spinal fusion but had no bracewear. Patients who were braced had a scoliosis that reached a magnitude of 50 degrees at a mean 12.5 years, compared with a mean age of 14 years for those who were not braced (p > 0.05). Spinal orthotics had no impact on scoliosis curve, shape, or rate of progression in spastic quadriplegic patients who were followed-up on fusion. Apical vertebral rotation > 2 (Nash and Moe criteria) is an indicator of impending rapid progression of the scoliosis curve. PMID- 8728633 TI - Severe lumbar lordosis after dorsal rhizotomy. AB - Two children with spastic quadriplegia who developed excessive lumbar lordosis after selective dorsal rhizotomy are described. The rhizotomy did not change the ambulatory status of either child (one nonambulator, one household ambulator). Preservation of unopposed hip flexion in the presence of multiple laminectomies may lead to the development of a lordotic deformity in children who sit most of the time. Excessive lumbar lordosis may cause pain and difficulty in sitting. Surgical correction of this deformity is complex because of the removal of posterior elements during the rhizotomy. PMID- 8728634 TI - Skull thickness and halo-pin placement in children: the effects of race, gender, and laterality. AB - To study the effects of gender, race, and laterality on skull thickness, we reviewed 41 pairs of head computed tomography (CT) scans, matched for age and sex by race. Thickness was measured at the standard locations for halo pins (anterolateral [AL] and posterolateral [PL]) using bone windows. The average measurements ( +/- 1 SD) were right AL = 2.9 +/- 1.0, left AL = 2.8 +/- 1.0, right PL = 3.2 +/- 1.2, and left PL = 3.6 +/- 1.2 mm (range, 1.2-7.2). There were no statistically significant differences by race or gender. The average thickness increased with age. There was a difference between the right and left PL sites (3.2 +/- 1.1 vs. 3.6 +/- 1.2 mm) but not between the AL sites. Caution in halo application must be applied equally to all children. PMID- 8728635 TI - Abnormalities of the spine in Goldenhar's syndrome. AB - The vertebral abnormalities present in 35 patients diagnosed with Goldenhar's syndrome are reported. There was no absolute correlation between the presence of any of the formative or segmentation vertebral defects and that of other concomitant malformations. This study supports the conclusion that patients with various groupings of Goldenhar-related anomalies should be considered as a single entity, to which the name Goldenhar association could be applied. PMID- 8728637 TI - A prospective evaluation of idiopathic left thoracic scoliosis with magnetic resonance imaging. AB - Twenty-nine consecutive patients with idiopathic left thoracic scoliosis were prospectively studied using magnetic resonance imaging (MRI). T1-weighted sagittal and axial images were obtained on all patients from the brainstem to the tip of the conus. Two patients (7%) had a syrinx on MRI. The remaining 27 patients had normal MRIs. The prevalence of brainstem and spinal cord anomalies was much less common than reported in previous retrospective reviews. PMID- 8728636 TI - Routine preoperative MRI and SEP studies in adolescent idiopathic scoliosis. AB - In this prospective study, 72 patients with the clinical diagnosis of adolescent idiopathic scoliosis underwent routine preoperative magnetic resonance imaging (MRI) scans and neurologic consultations. Forty-eight patients also had preoperative somatosensory evoked potentials (SEPs). All patients had normal neurologic examinations. Abnormal findings included two patients with Chiari type I malformation and one with a finding of a fatty collection in a vertebral body. In four cases, interpretation of the MRI was suspicious or equivocal, necessitating a computed tomography myelogram or other additional studies for clarification. Abnormal preoperative SEP results were obtained in three patients, none of which proved significant. All surgical patients underwent instrumentation and fusion without incident. The results indicate that routine preoperative SEP is not necessary. Routine preoperative MRI is probably not indicated in adolescent idiopathic scoliosis if the patient has a normal neurologic examination. PMID- 8728638 TI - Flexion and extension cervical MRI in a pediatric population. AB - Cervical spine flexion/extension magnetic resonance imaging (MRI) was used in 17 pediatric patients (age, 9 months to 12 years) for assessment of spinal cord compression secondary to instability. Nine of these patients had cord compression demonstrated on flexion/ extension MRI (flexion and extension, five; flexion, three; extension, one). These nine patients all had neurologic manifestations and underwent surgery. Patients without spinal cord compression on MRI did not demonstrate any neurologic manifestations. Flexion/extension MRI of the cervical spine was very useful in selecting appropriate patients for arthrodesis and determining the necessity for concurrent anterior or posterior decompression. PMID- 8728639 TI - Competitive sports and the progression of spondylolisthesis. AB - To consider the effects of several years of competitive sports training on children and adolescents with spondylolisthesis, we carried out a retrospective radiologic and clinical study of 86 young athletes with spondylolysis or spondylolisthesis (24 girls and 62 boys between the ages of 6 and 20 years). The mean degree of displacement was 10.1% at the beginning of the observation. The radiologic tests showed an increase in displacement over time in 33 athletes. The average progression of spondylolisthesis in this group was 10.5%. For 36 athletes, spondylolisthesis did not progress during the period of athletic training. In seven athletes, a decrease in the displacement was observed, from 17.9 to 8.9% on average. For 10 athletes, the course of spondylolisthesis could not be determined, because only one lateral radiograph was available. In spite of intensive daily training, the athletes had no symptoms during the entire period of observation, which lasted an average of 4.8 years. In light of our experiments, there is no justification for generally advising children and adolescents with limited spondylolytic spondylolisthesis not to take part in competitive sports. PMID- 8728640 TI - Nicardipine for controlled hypotension during spinal surgery. AB - Nicardipine is the first intravenously administered dihydropyridine calcium channel blocker. Its primary physiologic actions include vasodilatation with limited effects on the inotropic and dromotropic function of the myocardium. Several reports have documented its use in adult patients for pharmacologic control of blood pressure. We present our experience with nicardipine as an agent for controlled hypotension during spinal surgery in 24 children. After the induction of general anesthesia, nicardipine was started at 5 (22 patients) or 10 micrograms/kg/min (two patients). The target mean arterial pressure (MAP) of 55 65 mm Hg was reached in 5.1 +/- 2.1 min (range, 2-10). Intraoperative infusion requirements to maintain the target MAP varied from 0.5 to 7 micrograms/kg/min (mean, 2.5 +/- 1.1). No adverse effects related to nicardipine were noted. Nicardipine appears to be an effective agent for controlled hypotension in children. Future studies are required to determine its advantages/disadvantages compared with more commonly used agents such as sodium nitroprusside or adrenergic antagonists. PMID- 8728641 TI - The safety of continuous epidural infusion for postoperative analgesia in pediatric spine surgery. AB - Epidural analgesia and anesthesia are standard regional techniques in orthopaedic surgery of the lower extremities. Benefits of epidural anesthetic infusions include excellent analgesia, minimal respiratory depression, no somnolence, and decreased need for blood transfusion. Adverse effects include pruritus, nausea, and urinary retention, but standard methods have evolved to counter each adverse effect. A continuous epidural infusion of opioid and bupivacaine was used as the principal postoperative analgesic for 71 young patients undergoing surgery for the correction of spinal deformity. The infusion was titrated to a point at which each patient denied having any pain and was maintained for an average of 2.9 days. Sixty-four patients experienced satisfactory analgesia with minimal adverse effects. The technique worked despite multiple laminotomies for segmental fixation and did not compromise neurologic assessment. We conclude that epidural analgesia is as safe and effective after spinal-deformity surgery as it is after other types of surgery. PMID- 8728642 TI - An evaluation of the posterior leaf spring orthosis using joint kinematics and kinetics. AB - The primary function of the posterior leaf spring orthosis (PLS) is to prevent excessive equinus or drop foot in swing. The name of the orthosis, posterior leaf "spring," suggests that it also mechanically augments push-off in stance. The purpose of this study was to determine the effect of the PLS on ankle function by using computerized gait-analysis techniques. Multiple barefoot versus brace walks were compared in 31 children with cerebral palsy. Results indicate that the PLS reduces excessive equinus in swing and is sufficiently flexible to allow ankle dorsiflexion in midstance. In terminal stance, the peak power-generating capabilities of the ankle were reduced when the child was wearing the PLS. Energy results indicate that more mechanical energy was absorbed during midstance and less produced during terminal stance with the PLS. Therefore, the PLS improved ankle function but did not augment ankle function through storage and return of mechanical, or spring, energy. PMID- 8728643 TI - A simple technique for assessing heel contact in orthoses. AB - Orthoses are used by patients with neuromuscular disorders to control their ankle and foot position. Heel contact inside an orthosis is difficult to determine. A simple, inexpensive device was constructed that could be positioned between the heel and the inside of the orthosis. It consists of a thin, single-cell pressure sensor connected to a low-cost, digital multimeter. The resistance reading was used to determine the presence or absence of heel contact during a controlled standing protocol. The device was evaluated in 23 patients. The majority of patients did not weightbear on their heels. The heel contact of five patients was reassessed by using a computerized F-scan system, which determines plantar weightbearing forces. A 100% agreement was found between the single-cell sensor and the F-scan system. This single-cell sensor is a simple, inexpensive, and easily used device to determine the presence or absence of heel contact within an orthosis. PMID- 8728644 TI - Free gracilis muscle transfer for coverage of severe foot deformities. PMID- 8728646 TI - Viruses. AB - The structures of the components of large and complex viruses, determined over the past year, have demonstrated the great variation in the ways in which viruses achieve their goals. The structure of the bluetongue virus coat protein provides clues as to how a T = 13 particle is assembled and the structure of the tick borne encephalitis envelope protein suggests a new way of exposing a membrane fusion peptide at the right moment. PMID- 8728645 TI - Circular assemblies. AB - During 1994 and 1995, the structures of the serum amyloid P component, the bacterial chaperonin GroEL, the 20S proteasome, the bacterial light-harvesting complexes and the tryptophan operon RNA-binding attenuation protein have been determined. These structures all form circular assemblies in which the individual subunits are related by rotational symmetry. In most cases the circular organization generates a new biophysical property and a specific biological function which have presumably been selected by evolution. PMID- 8728647 TI - Assembly of macromolecular complexes in bacterial and baculovirus expression systems. AB - Many proteins exist normally as oligomers or complexes with other proteins. Recent advances in vector design have allowed this aspect of protein function to be mimicked in recombinant expression systems. Examples of the ordered oligomerization of a single protein through to the assembly of eight different proteins have been documented in recombinant Escherichia coli and recombinant baculovirus systems. PMID- 8728648 TI - Nuclear pores and macromolecular assemblies involved in nucleocytoplasmic transport. AB - Transport in and out of the nucleus is mediated by nuclear pore complexes (NPCs). Information has recently been emerging both on the structure of many of the proteins involved in this transport and on the complexes formed between them. In addition to NPC-based complexes, such as those based on nucleoporins Nsp1p and p62, cytoplasmic protein macromolecular assemblies are also important for nucleocytoplasmic transport. PMID- 8728649 TI - The use of hydrostatic pressure as a tool to study viruses and other macromolecular assemblages. AB - Recent studies on the effect of pressure on macromolecular assemblages have provided new information on protein-protein and protein-nucleic acid interactions. New findings have recently emerged on the use of hydrostatic pressure to assess intermediate states in the assembly pathways of viruses, multimeric proteins and protein-nucleic acid complexes, addressing many questions of macromolecular recognition. PMID- 8728650 TI - Use of macromolecular assemblies as expression systems for peptides and synthetic vaccines. AB - The past decade has witnessed the development of numerous systems for the presentation of antigens on the surface of self-assembling macromolecules. Although the sites for insertion were initially chosen empirically, the determination of the three-dimensional structures of a number of carrier macromolecules has enabled structure-based insertional mutagenesis to be used increasingly. Furthermore, it is now feasible to determine the structure of an inserted sequence as presented in a heterologous environment, making it possible to correlate the detailed structure of a peptide with its immunological properties. PMID- 8728651 TI - Structure of gap junction intercellular channels. AB - Gap junctions are formed by a multigene family of polytopic membrane channel proteins, connexins, that have four hydrophobic transmembrane domains and their N and C termini located on the cytoplasmic membrane face. The C-terminal tail plays important roles in channel regulation by pH and phosphorylation. Conserved cysteine residues stabilize the conformation of the extracellular loops that mediate the 'docking' between connexons in the intercellular channel. Over the past year, electron cryocrystallography of two-dimensional crystals of a truncated recombinant alpha 1 (Cx43) has revealed that the transmembrane boundary of the intercellular channel is lined with alpha helices. Furthermore, a ring of alpha helices resides at the interface with the membrane lipids. A three dimensional analysis based on images recorded from tilted crystals should reveal the location and secondary structure of additional transmembrane domains, as well as provide important structural details about the interactions between connexins within a hemi-channel and connexon-connexon interactions in the extracellular gap. PMID- 8728652 TI - Structure-derived potentials and protein simulations. AB - There has recently been an explosion in the number of structure-derived potential functions that are based on the increasing number of high-resolution protein crystal structures. These functions differ principally in their reference states; the usual two classes correspond either to initial solvent exposure or to residue exposure of residues. Reference states are critically important for applications of these potentials functions. Inspection of the potential functions and their derivation can tell us not only about protein interaction strengths themselves, but can also provide suggestions for the design of better folding simulations. An appropriate goal in this field is achieving self-consistency between the details in the derivation of potentials and the applied simulations. PMID- 8728653 TI - Potential energy functions for threading. AB - Despite little progress in ab initio solutions to the problem of predicting a protein's tertiary structure, over the past four years or so the development of fold-recognition methods for tertiary structure prediction has been the source of some encouragement in this difficult field. Despite promising initial results, these methods are clearly not yet fully mature and many groups are now working on different aspects of the methods involved in the hope of increasing the reliability and sensitivity of these tools. PMID- 8728654 TI - Modeling side-chain conformation. AB - Over the past few years, a number of methods for the calculation of side-chain conformations in proteins have been described. More recent studies have considered the effect of combinatorial packing, derivations from idealized rotameric structures and, to a limited extent, backbone flexibility on the quality and efficiency of calculations of protein side-chain conformation. Although further work is needed to address the issue of backbone displacements, the recent progress solves the packing problem to a significant degree. This opens the way for fruitful incorporation of these methods into general procedures for homology modeling and studies of ligand-protein interactions. PMID- 8728655 TI - Mean-field minimization methods for biological macromolecules. AB - Simulations of macromolecular structures involve the minimization of a potential energy function that presents many local minima. Mean-field theory provides a tool that enables us to escape these minima, by enhancing sampling in conformational space. The number of applications of this technique has increased significantly over the past year, enabling problems with protein-homology modelling and inverted protein structure prediction to be solved. PMID- 8728656 TI - Genetic algorithms for protein structure prediction. AB - Genetic algorithms are a general class of search methods that mimic natural gene based optimization mechanisms. Mutation, cross-over and replication operations are performed on strings. When applied to structure prediction, each string describes a particular conformation of a protein molecule. There are many ways in which such search methods may be implemented. Recent results show potential for helping with protein structure prediction, but more data are needed before a complete assessment can be made. PMID- 8728657 TI - Novel methods for molecular dynamics simulations. AB - In the past year, significant progress was made in the development of molecular dynamics methods for the liquid phase and for biological macromolecules. Specifically, faster algorithms to pursue molecular dynamics simulations were introduced and advances were made in the design of new optimization algorithms guided by molecular dynamics protocols. A technique to calculate the quantum spectra of protein vibrations was introduced. PMID- 8728658 TI - Modeling protein-ligand complexes. AB - Increasing the rate at which new biologically active compounds are found is a major goal in pharmaceutical chemistry. Recently, several computational methods have been proposed with this intent. For some time, algorithms have been used to direct ligand evolution on the basis of complementarity to the three-dimensional structure of a selected protein. Current research focuses on enhancements to methods for searching chemical databases, proposing sensible modifications to known active compounds, and construction of novel ligands from theoretical principles. PMID- 8728659 TI - Simulating DNA at low resolution. AB - The past year has witnessed the development of several new mathematical approaches to analyzing the structure of double-helical DNA and to incorporating the sequence-dependent features of the chain in computer simulations of long polymers. Of special interest in this respect are the local and global structural changes induced by the binding of various proteins to DNA, ranging from subtle bending, untwisting and sliding motions at the base-pair level to the apparent organization of supercoiled structure in chains that are thousands residues long. The computational effort has also included both new ways to incorporate the polyelectrolyte character of DNA and other environmental forces in simulations of long chains and new methods to keep track of the multitude of configurations so generated. The collective advances are pointing to ways that will soon connect the sequences of base pairs in large genomes to folded three-dimensional structures based on natural bending, twisting and translational tendencies and in response to deformations produced by the binding of different proteins. PMID- 8728661 TI - Theory and simulation. PMID- 8728660 TI - Macromolecular assemblages. PMID- 8728662 TI - The value of studying occupation: an example with primate play. AB - The purpose of this article is to demonstrate the value of a constraint-free study of occupation to occupational therapy. Constraint-free study refers to interdisciplinary scholarly inquiry that seeks to generate a deep understanding of how important occupations influence adaptation and health. Drawing on knowledge from animal behavior, anthropology, evolutionary biology, psychology, and primatology, primate play is discussed. Such knowledge enhances understanding of therapeutic play contexts and of play's integrative impact on occupational performance. Moreover, a robust understanding of vital occupations, such as play, empowers practitioners, thereby reducing the profession's vulnerability to external forces that diminish its economic and political position in the health care market. Precedents within and outside occupational therapy are described whereby control over the acquisition of basic knowledge has proven to be beneficial. It is concluded that both occupational therapy and occupational science will strongly benefit from their shared commitment to understanding the complexities of occupation. PMID- 8728663 TI - Metaphor and meaning in a clinical interview. AB - This study examined the narrative features of 20 life histories gathered from psychiatric patients with the Occupational Performance History Interview. The aim was to identify how narrative features were present in the patient interview responses and to illustrate how such narrative features can be located. We found that the patients organized their interview responses with deep metaphors that served to "emplot," or give meaning to, the life story. This article illustrates how patients used the deep metaphors to both circumscribe and frame possible solutions to the problems in their lives. Deep metaphors are consistent, recurring images of a life story that give coherence to, and aid in, the interpretation of the events of that life. Moreover, we explored how metaphors can be located in patient life histories and their implications for occupational therapy. PMID- 8728664 TI - Soft versus hard resting hand splints in rheumatoid arthritis: pain relief, preference, and compliance. AB - OBJECTIVES: This study compared soft versus hard resting hand splints on pain and hand function in 39 persons with rheumatoid arthritis. Splint preference was also evaluated to determine its effects on splint wear compliance. METHOD: A repeated measures research design was used to compare the two experimental conditions, wearing a soft splint versus a hard splint on the dominant hand for 28 days at night only, and an unsplinted control period of 28 days. RESULTS: Arthritis pain was considerably less during both splinted periods when compared with the pretest. Subjects identified fewer joints as being painful during the soft splint condition than during the unsplinted condition. There were no significant differences among conditions on hand function measures. Splint preference was 57% for the soft splint, 33% for the hard splint, and 10% for no splint. Splint wear compliance was significantly better with the soft splint (82%) than with the hard splint (67%). CONCLUSION: The findings indicate that resting hand splints are effective for pain relief and that persons with rheumatoid arthritis are more likely to prefer and comply with soft splint use for this purpose. Individualized splint prescription that focuses on client comfort and preference may enhance splint wear compliance. PMID- 8728665 TI - Minimal forces to move patients. AB - OBJECTIVES: Health care workers who have patient transfer duties are at risk for back injury. Transferring patients between beds and gurneys is a rigorous pulling task that requires back, leg, and arm strength. This study analyzed the efficiency of commercially available transfer devices, namely a patient roller, patient shifter, and draw sheet. METHOD: With the use of one or two force scales attached to each transfer device, the amount of force needed to transfer 15 participants, weighing from 101 lb to 240 lb, back and forth between a bed and a gurney was measured. Ten transfers per device per participant were performed. RESULTS: The patient roller was superior to the patient shifter and draw sheet in reducing transfer forces. Additionally, gurney-to-bed transfers tended to be more demanding for all transfer devices and for heavier participants. CONCLUSIONS: The patient roller was the most efficient transfer device in moving participants compared with the draw sheet and patient shifter. Transfer forces can be estimated with the use of linear equations, with patient weight, direction of transfer, and transfer device as the independent variables. These estimated forces can assist occupational therapists in the returning their injured health care coworkers to patient transfer duties. The results further indicated that high forces are required to transfer patients; therefore, patient-transfer personnel should obtain assistance when moving patients. PMID- 8728666 TI - Use of electronic music as an occupational therapy modality in spinal cord injury rehabilitation: an occupational performance model. AB - This article describes an electronic music program that allows clients with spinal cord injury (SCI) to form musical bands and play songs while performing therapeutic exercise in an occupational therapy program. Clients create the music by activating upper extremity exercise devices that are connected to a synthesizer through a computer. The bands choose the songs they would like to play and meet twice a week for 1 hr to practice. The 8-week program often concludes with a public performance. The music program is intended to motivate client participation in physical rehabilitation while promoting self-esteem, emotional expression, and peer support. It is based on the model of occupational performance and the theory of purposeful activity. To date, 33 persons have taken part. Client, therapist, and public response has been positive because this program highlights the abilities of persons with SCI, thereby encouraging their reintegration into the community. PMID- 8728667 TI - Perceptions and experiences of occupational therapists in rural schools. AB - Occupational therapy practice in rural schools is influenced by a number of factors unique to rural settings, such as geographic barriers, cultural diversity, limited access to external resources, and lack of personnel with specialist skills. The purpose of this ethnographic study was to identify the unique experiences of occupational therapists who practice in rural schools and to explore how they perceived their roles in rural schools. Six occupational therapists were interviewed and observed in the field. Analysis of interview data revealed five themes: (a) jack-of-all-trades, which described how the participants viewed themselves as generalists; (b) bridging the span between services, which described their roles in coordinating the students' medical services, which tended to be disjointed; (c) the world can get kind of lonely out there, which described the sources of support seen as reducing their sense of isolation; (d) trust and teaming, which described their attempts to provide an integrated intervention service and the challenges they faced; and (e) I cannot do it all, but I wish I could, which described how they selected their service delivery models to meet the demands of their practice. The participants believed that their work in rural schools required creativity, resourcefulness, and self reliance. These characteristics, as well as support from their colleagues, were important to coping with the challenges of practice in rural schools. PMID- 8728668 TI - Occupational therapy task observation scale (OTTOS): a rapid method for rating task group function of psychiatric patients. AB - OBJECTIVE: We designed a simple rating instrument, the Occupational Therapy Task Observation Scale (OTTOS), to facilitate evaluation and documentation of patient performance during occupational therapy task groups and to improve the communication between occupational therapists and other treatment team members. METHOD: After analysis of other rating instruments, a preliminary version of OTTOS was designed, extensively field tested, and further modified. Interrater reliability was determined, and validity was tested against three established rating instruments: the Bay Area Functional Performance Evaluation, the Comprehensive Occupational Therapy Evaluation Scale, and the Milwaukee Evaluation of Daily Living Skills. RESULTS: The final version of OTTOS contains two parts, 10 items for evaluation of specific task functions and 5 items for rating general behavior. Field use demonstrated that the scale successfully tracked changes in functional capacity and included most facets of patient function evaluated during task groups. Use of OTTOS required minimal training, and scoring required less than 2 min for each patient. The correlation between the scores of experienced occupational therapists was high (.92 for the total scores). The correlation between OTTOS and the other rating instruments ranged from .880 to .340; the highest correlations, as expected, were with test subscales that most closely resembled OTTOS: CONCLUSION: Preliminary data indicated that OTTOS offers a reliable and valid method for rapidly rating the function of patients with psychiatric illness attending task groups. In addition, use of OTTOS improved the communication between occupational therapists and other health care providers, facilitated the education of occupational therapy students, and met the documentation requirements of third-party payers. PMID- 8728669 TI - The use of the edema bar for the treatment of burn patients. AB - The edema bar is a new device that therapists can add to their repertoire of treatment approaches for burn patients. This article looks at the challenges therapists encounter while working with burn patients and describes the edema bar's design, its application to the burn rehabilitation process, and the multidisciplinary approach to its use. PMID- 8728670 TI - Is the use of life history and narrative in clinical practice reimbursable? Is it occupational therapy? PMID- 8728671 TI - Is the use of life history and narrative in clinical practice fundable as research? PMID- 8728672 TI - Physiology and evaluation of ureteropelvic junction obstruction. AB - Ureteropelvic junction (UPJ) obstruction is not a single anatomic entity but rather a set of processes with multiple causes. In the obstructed system, histologic changes are apparent that have implications for healing. The role of crossing vessels In UPJ obstruction and its treatment is of growing interest. Various methods are available to assess the functional characteristics of the UPJ, of which the diuretic renogram, Whitaker perfusion test, and Doppler ultrasound resistive index may be the most useful. The treatment must be individualized according to the patient's symptoms, the status of renal function, and the potential complications of intervention. PMID- 8728673 TI - Imaging for ureteropelvic junction obstruction in adults. AB - The three goals of imaging for ureteropelvic junction (UPJ) obstruction are to determine the presence and degree of renal obstruction, to assess renal function, and to determine the cause of the problem. The diagnostic techniques are intravenous urography, which is unreliable; ultrasonography, with the addition of pulsed Doppler technology; diuretic scintigraphy; and pressure-flow studies (Whitaker test), which is the only direct clinical test of renal outflow resistance. Voiding cystography, retrograde ureteropyelography, and CT have utility in some patients. In determining the cause of obstruction, the patient often is evaluated for crossing vessels using angiography, endoluminal ultrasonography, or spiral CT. It is not possible to formulate a single algorithm for radiographic evaluation of suspected UPJ obstruction, but in the average adult patient, urography and diuretic scintigraphy are sufficient. PMID- 8728674 TI - Endoluminal sonographic imaging of the ureteropelvic junction. AB - Endoluminal ultrasound imaging of the ureter can demonstrate the periureteral anatomy. Ultrasound probes are available as catheters from 3.5F to 6.2F. The transducer is available as a 12.5 or 20 MHz design and gives a cross-sectional image with penetration of 1.5 to 2 cm. Endoureteral ultrasonography has demonstrated blood vessels associated with the obstructed ureteropelvic junction (UPJ) in more than 50% of patients. These vessels have been located at all sites adjacent to the UPJ. The anteromedial location is the most common site. In comparison, only 8 significant crossing vessels were found in 44 patients with a normal UPJ. Inspection of the UPJ with a high insertion of the ureter demonstrates a characteristic sonographic image with a septum between the ureteral and renal pelvic lumen. Endoluminal ultrasonography of the obstructed UPJ can be used to guide the site of endopyelotomy to avoid damage to adjacent vessels. The presence of such vessels may also be related to the success of the procedure. An ultrasound-guided cutting device has been developed for preclinical studies. PMID- 8728675 TI - Vessels around the ureteropelvic junction: significance and imaging by conventional radiology. AB - We studied the influence of vessels crossing the ureteropelvic junction and of the grade of hydronephrosis in 86 patients who underwent endoureteropyelotomy. With a follow-up ranging from 1 to 12 years, the presence of crossing vessels had a significant negative influence on the outcome and recurrence rate of the procedure. The success rate was 82% when no vessels were present but only 33% when crossing vessels were documented. In 13 of 15 failures (87%), a crossing vessel was found at open pyeloplasty. The degree of hydronephrosis was a less potent detrimental factor, with an 81% success rate in low-grade and 54% in high grade hydronephrosis. The best results (90% success) were obtained in patients with no crossing vessels and low-grade hydronephrosis and the worst (30% success) in those with high-grade hydronephrosis in the presence of crossing vessels. Selection of patients according to those prognostic factors is recommended. Conventional intravenous urographic imaging techniques proved insufficient, and only angiography provided diagnostic accuracy; less-invasive methods are desirable. PMID- 8728676 TI - Percutaneous endopyelotomy and results: Korth technique. AB - Endoscopic incision of ureteral strictures has become an important component of the urologic armamentarium. Despite different techniques and stenting times applied for the procedures, the rates of long-term favorable outcomes have proved to range consistently from 72% to 89% for both secondary and primary hydronephrosis. Cold-knife incision combined with 3 weeks of stenting has been utilized at our institution from the beginning. The results have been as favorable as those from other groups. However, stenting times are the subject of controversy. In a prospective trial, a newly designed stent for internal reflux free drainage and subcutaneous fixation was applied in 53 patients and left indwelling for 6 months. Whereas such long-term stenting using the Endostent seemed to produce less favorable overall results, stenting for 3 weeks proved sufficient in lower-grade hydronephrosis secondary to small stenotic ureteral segments. Although stenoses recurred in 12% of our cases, 90% of these recurrences could be treated endoscopically. All but two recurrences became evident within the first 6 months after stent removal. After a mean follow-up of 20.2 +/- 19.6 months, the outcomes have remained unchanged even on late reexamination. PMID- 8728677 TI - Antegrade transpelvic endopyelotomy in primary obstruction of the ureteropelvic junction. AB - The authors present a 5-year follow-up of endopyelotomy using a personal technique for cases of primary ureteropelvic junction obstruction. After percutaneous access has been gained via a lower calix, the technique involves wide opening of the renal pelvis and exploration of the peripelvic space before a 3- to 4-cm long sectioning of the ureter. The aim is to carry out all of the operation in full view and without the need for a large-caliber stent, in order to perform endopyelotomy also in pediatric patients or in presence of anomalous vessels. The follow-up demonstrates a good result in 80% of 46 patients aged 5 to 62 years; two patients underwent surgical repair. The authors think the antegrade transpelvic endopyelotomy is an endourologic operation whose results and feasibility parallel those of open surgery. PMID- 8728678 TI - Acucise endopyelotomy: evolution of a less-invasive technology. AB - Since its introduction in 1993, the electrosurgical cutting balloon device (Acucise) has been utilized for endopyelotomies by a number of investigators. The fact that the procedure can be performed in a completely retrograde fashion under fluoroscopic guidance without percutaneous access has made Acucise endopyelotomy appealing to many urologists. To date, overall Acucise endopyelotomy success rates ranging from 66% to 84% have been reported. The average hospital stay has ranged from 1.7 to 3.7 days, and serious complications have been rare (transfusion 0-2%, bleeding necessitating embolization 0-3%). On the basis of its track record and its minimally invasive nature, a compelling argument can be made for use of the Acucise device whenever an endopyelotomy is indicated. PMID- 8728679 TI - Ureteroscopic retrograde endopyelotomy for management of ureteropelvic junction obstruction. AB - Despite the success of endopyelotomy, many points are still being debated. Ureteroscopic retrograde endopyelotomy was carried out in 49 adult patients with 39 available for long-term follow-up. Prestinting facilitated instrument passage and reduces morbidity. The average operating time was 90.2 minutes. Two patients underwent elective nephrectomy for poorly functioning kidneys that did not improve after treatment, and one patient underwent urgent nephrectomy for bleeding 26 days postoperatively. Improvement in drainage was documented in the successful cases. The authors consider ureteroscopic endopyelotomy the first-line approach for most cases of ureteropelvic junction obstruction. PMID- 8728680 TI - New technique in managing ureteropelvic junction obstruction: percutaneous endoscopic pyeloplasty. AB - Endopyelotomy is currently advocated for management of the obstructed ureteropelvic junction (UPJ). Healing of the stented UPJ occurs by secondary intention. Regardless of the method employed, success rates approach 85%. In order to increase the rate of success, we have devised a method of performing endopyelotomy in conjunction with endoscopic suturing of the incised UPJ. Two methods were developed to allow for the placement of a single absorbable monofilament suture. In the first method, endopyelotomy is carried out in the standard antegrade manner, and endoscopic suturing is performed with the use of a second retroperitoneal access sheath at the UPJ. In the second method, endoscopic placement of the suture is carried out through the standard renal access sheath, with suturing performed via the nephroscope. We have performed endoscopic pyeloplasty in eight patients. With a mean follow-up of 12 months, the procedure was successful in seven of these patients. Endoscopic suturing of the UPJ is technically demanding, but once sufficient expertise is gained, it may be utilized for immediate tissue coaptation, possibly decreasing urinary extravasation and, it is hoped, maximizing the caliber of the UPJ. PMID- 8728681 TI - Ureteropelvic invagination procedure for endopyelotomy (Gelet technique): review of 51 consecutive cases. AB - Between 1990 and 1995, we performed 51 endopyelotomies on 38 cases of primary and 13 of secondary obstruction of the ureteropelvic junction (UPJ) using the ureteropelvic invagination technique. Of the 51 patients in the series, 49 have been followed for a minimum of 3 months postoperatively (mean follow-up 16 months). Overall, success was achieved in 38 (77.5%). Endoscopic endopyelotomy was successful in 11 of 13 cases (84.5%) with secondary strictures. When the technique was used for the treatment of primary UPJ stricture, the success rate was only 75% (27 of 36). The presence of a crossing vessel was identified as the cause of failure in five cases of primary strictures; hence, we advocate the use of angiography to identify crossing vessels preoperatively. We recommend the use of the ureteropelvic invagination technique as the first-line therapy for primary hydronephrosis in adults in the absence of a crossing vessel. PMID- 8728682 TI - Laparoscopic pyeloplasty. AB - Thirteen patients underwent laparoscopic dismembered pyeloplasty at our institution. The operative time ranged from 4 to 8 (mean 5.4) hours with minimal blood loss. Two patients developed transient edema necessitating temporary drainage. The mean requirement for morphine postoperatively was 32 mg. At a mean follow-up of 13 months, all patients demonstrated both symptomatic and radiographic relief of obstruction. Laparoscopic dismembered pyeloplasty appears to be an effective minimally invasive alternative to treat symptomatic ureteropelvic junction obstruction. PMID- 8728683 TI - Laparoscopic division of crossing vessels at the ureteropelvic junction. AB - Endopyelotomy has become an accepted mode of treatment for primary and secondary ureteropelvic junction (UPJ) obstruction, but a 15% to 30% failure rate persists. The presence of crossing vessels at the UPJ has been implicated as a common cause of complications, failures, and recurrences. In the past, renal angiography was necessary to identify crossing vessels. We have utilized endoluminal ultrasonography to identify crossing vessels at the UPJ and to guide endoscopic incisional techniques. Previously, whenever crossing vessels were identified that could not be safely avoided during endopyelotomy, we had recommended dismembered pyeloplasty, an open surgical procedure with a long recovery time. We report our experience with laparoscopic division of crossing vessels in two patients, one with a symptomatic horseshoe kidney. Each patient had a large crossing vessel identified by endoluminal ultrasonography; consequently, endopyelotomy was abandoned. The location and distribution of the vessels were then delineated by angiography. The aberrant vessels were dissected free and divided laparoscopically. The patients returned to work within 1 week. Follow-up diuretic renal scans showed complete resolution of obstruction (T1/2 < 10 minutes) in one patient; no change was noted in the patient with a horseshoe kidney. Both patients have remained free of symptoms and normotensive for more than 12 months. Laparoscopic division of crossing vessels may play a role in the treatment of patients with extrinsic ureteral obstruction from aberrant vessels. PMID- 8728684 TI - Use of ureteral stents after endopyelotomy. AB - Despite the widespread practice of endopyelotomy for the management of ureteropelvic junction obstruction, the optimal conditions for ureteral healing after endoincision remain largely untested. Current recommendations for the use of large-caliber graduated endopyelotomy stents and a 6-week duration of stenting are being challenged by recent reports of successful outcomes with the use of standard, small-caliber ureteral stents and early stent removal. Moreover, improvements in stent design have led to the development of endopyelotomy stents with improved biocompatibility, enhanced case of insertion, and fewer adverse effects. PMID- 8728685 TI - Complications of endopyelotomy. AB - Complications of endopyelotomy can be classified as major (vascular, ureteral, or pulmonary injury or lost wire), minor (stent-related problems, infection, urinoma, or unrecognized incision of infundibular narrowing or renal pelvis), or failure to obtain the desired results. Awareness of the potential problems enables the surgeon to take preventative steps. PMID- 8728686 TI - Crossing vessels at the ureteropelvic junction: do they influence endopyelotomy outcome? AB - The majority of crossing vessels probably are inconsequential. Bleeding should not be a concern if the incision is made laterally. The optimal way to improve endopyelotomy results is to determine which crossing vessels truly are significant, and the authors describe the clinical trial that might be carried out to answer the question. PMID- 8728687 TI - Hypomelanosis of Ito and X;autosome translocations: a unifying hypothesis. AB - Hypomelanosis of Ito is a sporadic multisystem disorder known to be associated in many cases with chromosomal mosaicism. While no particular pattern is generally evident for the specific chromosomes involved in such patients, a subgroup of female patients exists in whom the common factor is the presence of a balanced, constitutional X;autosome translocation, with a cytogenetic breakpoint in the pericentromeric region of the X. It is argued here that the phenotype in these cases results not from the interruption of X linked genes but from the presence of mosaic functional disomy of X sequences above the breakpoint. PMID- 8728688 TI - Hypopigmented skin alterations resembling tuberous sclerosis in normal skin. AB - The significance of hypopigmented skin findings as manifestations of the gene for tuberous sclerosis (TS) in near relatives of TS patients is a difficult problem. We therefore studied the number and kind of whitish skin alterations found in 100 medical students and 100 school children. Ninety three percent of the former and 79% of the latter had some whitish lesions, many of them scars. Twenty percent of the adults and 12% of the children had roundish or oval macules larger than 10 mm in diameter, not known to be scars. In clinical practice with TS patients, our attention has been drawn to whitish raised masses or streaks in their first degree relatives. These were also found in study subjects so the significance of such lesions remains unclear. The role of Wood's light turned out to be questionable, far from pathognomonic for TS; 25% of all the whitish findings and 53% of the hypopigmented macules larger than 10 mm in diameter showed distinct or brilliant fluorescence under Wood's light. PMID- 8728689 TI - Ascertainment of familial ovarian cancer in the Aberdeen Genetic Clinic. AB - Ovarian cancer is the fifth most common malignancy in women in the UK. Patients with a family history including ovarian cancer make up nearly 15% of family cancer referrals to the Genetic Clinic in Aberdeen. To date, only one pedigree has been suitable for linkage studies, which has enabled us to target screening more accurately at those people at highest risk. Following discovery of a strong candidate for the BRCA1 gene, direct mutation testing may soon be possible. People who seek testing will require further counselling. Therefore we anticipate an increased demand on both clinical and laboratory resources, but more accurate ascertainment of high risk subjects should lead to more appropriate targeting of screening services. PMID- 8728690 TI - Arachnoid cyst and chronic subdural haematoma in a child with osteogenesis imperfecta type III resulting from the substitution of glycine 1006 by alanine in the pro alpha 2(I) chain of type I procollagen. AB - The features of a child with osteogenesis imperfecta type III (OI III) resulting from the heterozygous substitution of glycine 1006 by alanine in the pro alpha 2(I) chain of type I procollagen were studied. He was born at term with the clinical features of severe OI, including deep grey-blue sclerae. He had severe osteopenia and all long bones were smaller than normal with cortical thinning, metaphyseal expansion, poor metaphyseal modelling, and multiple fractures. However, the vertebrae, pelvis, and shoulder girdle were of normal shape and there were few rib fractures. Histological examination of the calvarium and tibial shaft showed woven bone without lamellar bone or Haversian systems. The shafts of the long bones were widened owing to repeated fractures. Progressive enlargement of the calvarium occurred between 3 and 4.5 months of age owing to bilateral chronic subdural haematomata and a large arachnoid cyst in the Sylvian fissure. The cyst was probably developmental in origin while the subdural collections were probably the result of perinatal skull trauma. The cyst and the subdural collections resolved following drainage but ventricular dilatation with normal cerebrospinal fluid pressure followed. The proband is the first reported case of OI with a glycine substitution by alanine in the pro alpha 2(I) chain of type I procollagen. PMID- 8728691 TI - Familial Wolf-Hirschhorn syndrome resulting from a cryptic translocation: a clinical and molecular study. AB - We present three cousins who have normal karyotypes, despite having clinical features of Wolf-Hirschhorn syndrome. Fluorescence in situ hybridisation techniques confirmed that all three relatives were monosomic for the distal short arm of chromosome 4 and that a cryptic translocation involving chromosomes 4 and 11 was segregating within the family. Segregation analysis indicated that the risk of an affected child being born to a parent carrying the translocation was 15%. Molecular analysis showed that loci D4S111 and D4S115 were not deleted in the proband, thus excluding these loci from the "Wolf-Hirschhorn critical region". Surprisingly, DNA studies also suggested that the translocation breakpoint on chromosome 4 was within the region of a preexisting paracentric inversion. PMID- 8728692 TI - Neonatal Schwartz-Jampel syndrome: a common autosomal recessive syndrome in the United Arab Emirates. AB - Schwartz-Jampel syndrome (SJS) is a heterogeneous autosomal recessive syndrome of myotonia and bone dysplasia. Two types have been recognised: the classical type with late infantile or childhood manifestation and a rarer form with neonatal manifestation. We report five families with a total of 11 children affected with severe neonatal SJS. All presented after birth with skeletal abnormalities and feeding difficulties. Five had the typical pursed appearance of the mouth. Nine died from respiratory complications (five in the neonatal period and four before 2 years of age). One (4 months old) remains hospitalised since birth requiring continuous oxygen supplementation and one (5 months old) requires nasogastric tube feeding and has repeated attacks of aspiration. Only seven of the 17 previously reported neonatal SJS cases had a similar course to the patients in this report. We suggest that within neonatal SJS there is a subgroup which manifests severe respiratory and feeding problems and has a poor prognosis. This report brings the total number of children with neonatal SJS reported from the UAE to 14. This represents the largest review of this syndrome to date from one centre and indicates that this syndrome is fairly common in the population of the UAE. PMID- 8728693 TI - Genetic referrals of Middle Eastern origin in a western city: inbreeding and disease profile. AB - Inbreeding or consanguineous marriage is a common traditional practice in Middle Eastern cultures. Studies from various countries and communities of this region showed that the frequencies range from 20% to greater than 70%. Inbreeding is known to have adverse effects on morbidity and mortality, in particular with respect to autosomal recessive disorders. This study examined 200 couples representing all referrals of Middle Eastern origin seen at a large Clinical Genetics Unit in Montreal. They were compared with a similar sized group of different cultural backgrounds from among the same referrals. The rate of intercultural marriages and inbreeding was found to be 24% and 23.5% respectively in the Middle Eastern group, while they were 22.5% and 5% in the comparison group. Excluding the referrals for consanguinity only, the rate of inbreeding among the study group was 16.4%. Within the Middle Eastern group, autosomal recessive disorders were more than twice as common in the inbred than in the non inbred families, the pattern of which is consistent with previous observations. PMID- 8728694 TI - X inactivation analysis in a female with hypomelanosis of Ito associated with a balanced X;17 translocation: evidence for functional disomy of Xp. AB - X inactivation analysis was performed on normal and hypopigmented skin samples obtained from a female with hypomelanosis of Ito associated with a balanced whole arm X;17 translocation. Severe skewing of X inactivation resulting in inactivity of the intact X was found in blood and cultures of both types of skin, but analysis of DNA prepared directly from hypopigmented skin showed significant inactivation of the translocated X, inconsistent with the usual mechanism of phenotypic expression in X;autosome translocations. In addition, dual colour FISH analysis using centromere specific probes for chromosomes X and 17 showed that the breakpoints on both chromosomes lie within the alphoid arrays, making interruption of a locus on either chromosome unlikely. While partial variable monosomy of loci on chromosome 17p cannot be excluded as contributing to the phenotype in this patient, it is argued that the major likely factor is partial functional disomy of sequences on Xp in cell lineages that have failed to inactivate the intact X chromosome. PMID- 8728695 TI - Frequency of myotonic dystrophy gene carriers in cataract patients. AB - DNA samples from 231 unselected patients with cataracts were studied to determine the frequency of the DM mutation in cataract patients. A previous epidemiological study established a high prevalence of DM in the population of Guipuzcoa (Basque Country, Spain), 26.5 cases/100,000. We have found two carriers (0.9%) of the DM mutation in patients who are not related to any previously known DM family. The screening of the DM mutation in cataract patients should be restricted to young patients or people with multicoloured and iridescent opacities, in which the risk of carrying the DM premutation could be higher. Our results suggest that subjects with 38 to 80 repeats could constitute the genetic reservoir of the DM mutation. PMID- 8728696 TI - Mental retardation, distinct craniofacial dysmorphism, and central nervous system malformation: confirmation of a syndrome. AB - We present a child with severe mental retardation, a central nervous system malformation, signs of ectodermal dysplasia, and a distinct craniofacial dysmorphism. Similar but less pronounced craniofacial features were present in the mildly mentally retarded mother. This observation confirms a previous report of a boy with the same MCA-MR syndrome and suggests X linked or autosomal dominant inheritance. PMID- 8728697 TI - Microcephaly, characteristic facies, joint abnormalities, and deficient leucocyte chemotaxis: a further case of the syndrome of Say et al. AB - We report on a 13 year old boy with microcephaly, sloping forehead, prominent nose, scoliosis, and flexion contractures involving the elbows and knees. The patient showed severe mental and growth retardation. Since birth and up to the present he has suffered from multiple and varied infections. Immunological studies showed a marked decrease in leucocyte chemotaxis. Clinical and laboratory findings confirm the similarity of this case to the two brothers described by Say et al. We have not found any descriptions of similar patients. The purpose of this paper is to contribute to the phenotypic delineation of this syndrome and to highlight the need for immunological investigation in patients with multiple congenital malformations. PMID- 8728698 TI - Odontomicronychial ectodermal dysplasia. AB - This paper describes odontomicronychial dysplasia, a pure ectodermal dysplasia of the 2-3 subgroup of group A. It is characterised by precocious eruption and shedding of deciduous dentition, precocious eruption of secondary dentition with short, rhomboid roots, and short, thin, slow growing nails. This condition probably results from an autosomal recessive gene. PMID- 8728699 TI - A second independent Tyr168Cys mutation in the tissue inhibitor of metalloproteinases-3 (TIMP3) in Sorsby's fundus dystrophy. AB - Sorsby's fundus dystrophy (SFD) is a rare autosomal dominant macular disorder with age of onset usually in the fourth decade. It is characterised by loss of central vision owing to subretinal neovascularisation and disciform macular degeneration. In an effort to identify the SFD gene, the disease locus was first mapped to chromosome 22q13-qter by genetic linkage analysis, the same chromosomal region as the gene encoding the tissue inhibitor of metalloproteinases-3 (TIMP3). Subsequently, two separate mutations in TIMP3 were found in affected members of two unrelated SFD pedigrees (Tyr168Cys and Ser181Cys). More recently, two additional SFD related mutations, Ser156Cys and Gly167Cys, have provided further confirmation that heterozygous mutations in TIMP3 are causally responsible for the SFD phenotype. We now report the occurrence of the Tyr168Cys mutation in an SFD patient of Austrian descent and show that this mutation found earlier in an American SFD family arose independently. The new findings add to an emerging pattern of SFD mutations which all seem to affect the C-terminal region of the mature TIMP3 protein. In addition, all known mutations cause a change of an amino acid to a cysteine residue. This suggests a critical role for the additional C terminal free thiol group in SFD pathogenesis. PMID- 8728700 TI - Cytogenetic and clinical characteristics of a case involving complete duplication of Xpter-->Xq13. AB - True isochromosomes for Xp probably do not exist in a liveborn. We describe a rare case of complete Xp duplication and retention of the inactivation centre at Xq13. Cytogenetically, it is described as a nonmosaic 46,X,psu idic(X)(q13). Complete duplication of Xpter-->Xq13 was confirmed by banded analysis and FISH probes for X centromere, Xp21, XIST locus, and whole chromosome paints for X and Y. The abnormal X was always late replicating. Clinically, the patient was short statured, had primary amenorrhoea, and incomplete development of secondary sexual characteristics, but otherwise was phenotypically normal. There are no non-mosaic reported cases with complete duplication of i(Xp) confirmed by FISH or molecular techniques. Those cases with partial duplication of Xp and presence of the inactivation centre share the traits of amenorrhoea and poor secondary sexual development. To develop a clinical profile of duplication of Xp (in presence of Xq13) there is a need to study more cases. PMID- 8728701 TI - A new deletion of 18q23 with few typical features of the 18q- syndrome. AB - We report on a patient with a deletion of 18q23. At both 2 and 4 years of age, she displayed few of the facial features or other clinical features associated with the 18q- syndrome. Fluorescent in situ hybridisation and microsatellite marker and RFLP analysis were performed to characterise the extent of the deletion, and a terminal deletion of 18q23 was confirmed. The deleted region includes the gene for myelin basic protein, suggesting that hemizygosity of this gene does not invariably lead to mental and developmental delay. The clinical presentation of this patient suggests that either she is not deleted for the genes involved in the 18q- clinical phenotype or this patient represents one end of the spectrum of the clinical variability seen with 18q terminal deletions. PMID- 8728702 TI - Presymptomatic diagnosis in Portuguese FAP families using intragenic RFLPs and (CA)n flanking markers by fluorescence based semiautomated DNA analysis. AB - Owing to the large size of the APC gene, responsible for familial adenomatous polyposis, direct screening for individual mutations is not a practical approach. In the present study we establish the methodology of fluorescence based semi automated DNA analysis to perform presymptomatic diagnosis of members at risk from 11 Portuguese FAP families with three (CA)n markers flanking the APC gene, MBC, CB26, and YN5.64, and four intragenic RFLPs. Haplotypes were constructed on the basis of individual genotypes and their segregation through generations were followed. The study was informative for 12% of subjects using only intragenic RFLPs and increased to 90% when we used the three (CA)n flanking markers. We report two of the 11 families under study in our laboratory and show recombinant events leading to a precise localisation of the CB26 marker between D5S82 and the APC gene. In one family there was a loss of (CA) units of one allele of the CB26 marker from an unaffected mother to her son. PMID- 8728703 TI - Discordant segregation of Xq28 markers and a mutation in the L1 gene in a family with X linked hydrocephalus. AB - X linked recessive hydrocephalus is the most common hereditary form of hydrocephalus. Genetic analysis indicates that the majority of cases are caused by mutations in a single gene in Xq28, recently identified as the gene for neural cell adhesion molecule L1. Genetic heterogeneity for this disorder was suggested following the description of a single large pedigree where X linked hydrocephalus showed lack of linkage to Xq28 markers flanking the L1 gene. Mutation analysis in this family shows a single base pair deletion within the coding sequence of the L1 gene that would result in truncation of the mature protein. The nature of the mutation and its segregation with the disease through the pedigree indicate that it is the cause of X linked hydrocephalus in this family. These results are at odds with data obtained through segregation of alleles for markers flanking the L1 gene. Somatic and germline mosaicism is the most plausible explanation for these data, which also provide further evidence for genetic homogeneity of X linked hydrocephalus. PMID- 8728704 TI - Molecular analysis of the arylsulphatase A gene in late infantile metachromatic leucodystrophy patients and healthy subjects from Italy. AB - A molecular analysis of the arylsulphatase A gene was performed on 26 unrelated, Italian, late infantile metachromatic leucodystrophy patients. The frequency of the common disease causing mutations 609A and 2381T was 28.8% and 1.9% respectively. Pseudodeficiency allele frequency in patients was found to be 13.5% and a frequency of 10.1% was found in 89 unaffected normal controls. The frequency of the 609A mutation in Italian late infantile patients is lower than in late infantile patients from northern Europe, suggesting a higher frequency of different sporadic mutations in the Italian population. A cooperative in cis effect in phenotype determination involving arylsulphatase A mutations and the eventual background of the pseudodeficiency allele is proposed. PMID- 8728705 TI - Molecular scanning of candidate mitochondrial tRNA genes in type 2 (non-insulin dependent) diabetes mellitus. AB - Mitochondrial DNA (mtDNA) gene defects may play a role in the development of non insulin dependent diabetes mellitus (NIDDM). In order to search for potentially diabetogenic mtDNA defects we have applied the technique of single stranded conformational polymorphism (SSCP) analysis to 124 patients with a history of NIDDM and 40 non-diabetic controls. No new heteroplasmic mutations were detected. However, a variety of homoplasmic variants were found in patients with NIDDM; some of these merit further investigation. PMID- 8728707 TI - Confirmation of linkage of Sjogren-Larsson syndrome to chromosome 17 in families of different ethnic origins. AB - Linkage analysis in two consanguineous pedigrees of Pakistani and English origin and one further Indian family in which affected subjects have Sjogren-Larsson syndrome (SLS) showed linkage to chromosome 17. Linkage of SLS to D17S783 and D17S805 has been reported in Swedish pedigrees, but since those data were generated from a single ethnic group originating from a common ancestor, there remained the question of whether this disease is genetically heterogeneous. This report confirms the linkage in non-Swedish pedigrees and, therefore, provides evidence to support a single locus for SLS. PMID- 8728706 TI - Clinical heterogeneity in hereditary haemorrhagic telangiectasia: are pulmonary arteriovenous malformations more common in families linked to endoglin? AB - Pulmonary arteriovenous malformations (PAVMs) occur in up to 27% of patients with hereditary haemorrhagic telangiectasia (HHT) and are associated with a rate of paradoxical cerebral embolism at presentation of up to 36%. At least two different loci have been shown for HHT. Mutations in endoglin have been found in some families and the locus designated ORW1. In other families this locus has been excluded. In this paper we confirm that in families linked to ORW1 there is a prevalence of PAVMs among affected members of 29.2%, compared to a prevalence of 2.9% in families in which this locus has been excluded (chi 2 = 19.2, p < 0.001). This information can be used to decide how to screen HHT patients for PAVMs. PMID- 8728708 TI - Monozygotic twins with chromosome 22q11 deletion and discordant phenotype. PMID- 8728709 TI - Large inv dup(15) chromosome in two generations. PMID- 8728711 TI - Two N-acetyl-D-galactosamine-specific lectins from Phaeolepiota aurea. AB - Two lectins, PAL-I and PAL-II, were isolated from the mushroom Phaeolepiota aurea by affinity chromatography on acid-treated Sepharose CL-4B followed by reverse phase FPLC on ProRPC. Both of the lectins were tetramers of 16 kDa subunits. The lectins had little agglutination activity against native erythrocytes but Pronase treatment of erythrocytes increased the sensitivity to agglutination by the lectins. Both lectins exhibited slight preferences for type A compared with type B and O erythrocytes. In haemagglutination inhibition assays, N acetylgalactosamine and both anomers of methyl N-acetylgalactosaminide were the best inhibitors. PMID- 8728710 TI - Simple tests for rhodopsin involvement in retinitis pigmentosa. PMID- 8728712 TI - Primary structure of a Kunitz-type trypsin inhibitor from Enterolobium contortisiliquum seeds. AB - A trypsin inhibitor was isolated from Enterolobium contortisiliquum seeds. Starting with a saline extract, ECTI (E. contortisiliquum trypsin inhibitor) was purified as a homogeneous protein by acetone precipitation, ion-exchange chromatography (DEAE-Sephadex A-50), gel filtration (Sephadex G-75 and Superose 12) and reversed phase HPLC (mu-Bondapak C-18). The amino acid sequence was determined by automatic degradation and by DABITC/PITC microsequence analysis of the reduced and carboxymethylated protein and also of purified peptides derived from the protein by cleavage with iodosobenzoic acid and by enzymic digestion with trypsin, chymotrypsin and Staphylococcus aureus V8 protease. ECTI contains 174 amino acid residues in two polypeptide chains, an alpha-chain consisting of 134 residues and a beta-chain made up of 40 residues. The inhibitor displays a high degree of sequence identity with other Kunitz-type proteinase inhibitors isolated from the Mimosoideae subfamily. The reactive site was identified (by homology) as the arginine-isoleucine peptide bond at position 64-65. ECTI inhibits trypsin and chymotrypsin in the stoichiometric ratio of 1:1 and also Factor XIIa, plasma kallikrein and plasmin, but not thrombin and Factor Xa. PMID- 8728713 TI - Purification and unusual kinetic properties of a tobacco anionic peroxidase. AB - The tobacco anionic peroxidase has been isolated from the leaves of transgenic Nicotiana sylvestris plants overproducing this enzyme. The plant expression system and the purification protocol developed allow the preparation of greater than 60 mg of homogeneous enzyme (M(r) 36 kDa, pI 3.5) from 1 kg of fresh leaves, which is an order of magnitude higher than for wild-type tobacco plants. The tobacco anionic peroxidase exhibits rather unusual catalytic properties in comparison with horseradish peroxidase (HRP C). Compound I is less active than Compound II in the tobacco enzyme. The enzyme is nearly inactive towards iodide, reflecting the peculiarities of its molecular structure. In particular, the presence of the negatively charged glutamate residue 141 at the entrance of the haeme-binding pocket seems to affect the stabilities of Compounds I, II and III, leading to a different enzyme substrate specificity than that of HRP C. Investigation of thermal stability towards a number of electron donors reveals the following 'order of stabilities': ferrocyanide > guaiacol > 2,2'-azino-bis(3 ethyl-6- benzothiazoline sulphonate) > iodide > o-dianisidine, which may indicate different binding sites and rate-limiting steps in the mechanism of the substrate oxidation. PMID- 8728714 TI - Antimicrobial steroids from the fungus Fomitopsis pinicola. AB - Phytochemical examination of the dichloromethane extract of the European fungus Fomitopsis pinicola led to the isolation of a new lanostanoid derivative identified from spectral and chemical evidences as 3 alpha-(4-carboxymethyl-3 hydroxy-3-methylbutanoyloxy)-lanosta++ +-8,24-dien-21-oic acid. In addition, seven known triterpenes, polyporenic acid C, 3 alpha-acetyloxylanosta-8,24-dien 21-oic acid, ergosta-7,22-dien-3 beta-ol,21-hydroxylanosta-8,24-dien-3-one, pinicolic acid A, trametenolic acid B and pachymic acid, were also isolated. Antimicrobial activity against Bacillus subtilis in a TLC bioassay was observed for five of the isolated steroids. PMID- 8728715 TI - Purification of (+)-delta-cadinene synthase, a sesquiterpene cyclase from bacteria-inoculated cotton foliar tissue. AB - A sesquiterpene cyclase whose activity is induced in a glandless, bacterial blight-resistant line of cotton (Gossypium hirsutum L.) catalyses the conversion of (E,E)-farnesyl diphosphate to (+)-delta-cadinene. This enzyme was purified by a combination of salt-induced phase separation, hydroxylapatite fractionation, hydrophobic interaction and strong anion-exchange chromatography, and denaturing polyacrylamide gel electrophoresis, followed by renaturation with Tween 80. The purified enzyme has a molecular weight of 64-65 kDa, and exhibited a single silver-staining band following electrophoresis in analytical denaturing polyacrylamide gels. Amino acid sequences of three tryptic peptides from the enzyme have been determined and are similar to known sequences in other terpene cyclases from plants. PMID- 8728716 TI - Saponins from Fagonia arabica. AB - Seven new triterpenoid saponins were isolated and identified from the aerial parts of Fagonia arabica. They were characterized as 3-O-beta-D-xylopyranosyl(1- >2)-[beta-D-glucopyranosyl(1-->3)]-alpha-L- arabinopyranosyl oleanolic acid 28-O beta-D-glucopyranoside, 3-O-beta-D-glucopyranosyl(1-->2)-[beta-D-glucopyranosyl(1 ->3)]-alpha-L- arabino pyranosyl oleanolic acid 28-O-beta-D-glucopyranoside, 3-O beta-D-xylopyranosyl(1-->2)-[beta-D-glucopyranosyl (1-->3)]-alpha- L arabinopyranosyl oleanolic acid, 3-O-beta-D-glucopyranosyl(1-->2)-[beta-D glucopyranosyl(1-->3)]-alpha-L- arabino pyranosyl oleanolic acid, 3-O-beta-D xylopyranosyl(1-->2)-[beta-D-glucopyranosyl (1-->3)]-alpha-L-arabinopyranosyl 27 hydroxyoleanolic acid, 28-O-beta-D-glucopyranoside, 3-O-beta-D-xylopyranosyl(1- >2)-[beta-D-glucopyranosyl(1-->3)]-alpha-L- arabinopyranosyl ursolic acid 28-O beta-D-glucopyranoside and 3-O-beta-D-xylopyranosyl (1-->2)-[beta-D glucopyranosyl (1-->3)-alpha-L-arabinopyranosyl 27-hydroxyursolic acid 28-O-beta D-glucopyranoside. The structures of the saponins were established by analyses of their 1H and 13C NMR spectra with the aid of 2D experiments. The two genins, 27 hydroxyoleanolic acid and 27-ursolic acid, are new. PMID- 8728717 TI - Triterpenoid saponins from Gymnema sylvestre. AB - Besides six known gymnemic acids, four new tritepenoid saponins, gymnemasins A, B, C and D, isolated from the leaves of Gymnema sylvestre, were identified as 3-O [beta-D-glucopyranosyl(1-->3)-beta-D-glucuronopyranosyl]-22-O- tigloyl- gymnemanol, 3-O-[beta-D-glucopyranosyl (1-->3)-beta-D-glucuronopyranosyl] gymnemanol, 3-O-beta-D-glucuronopyranosyl-22-O-tigloyl-gymnemanol and 3-O-beta-D glucuronopyranosyl-gymnemanol, respectively. The aglycone, gymnemanol, which is a new compound, was characterized as 3 beta, 16 beta, 22 alpha, 23, 28 pentahydroxyolean-12-ene. PMID- 8728718 TI - Cardenolides from Erysimum cheiranthoides. AB - Three new cardiac glycosides were isolated along with two known cardenolides from the seeds of Erysimum cheiranthoides. The new ones were characterized by spectral methods as 16 beta-hydroxystrophanthidin (strophadogenin) 3-O-beta-glucopyranosyl (1-->4)-beta-boiviopyranoside, strophadogenin 3-O-alpha-rhamnopyranosyl-(1-->4) beta-digitoxopyranoside and strophanthidin 3-O-alpha-rhamnopyranosyl-(1-->4)-beta digitoxopyranoside, named cheiranthosides I, II and III, respectively. PMID- 8728719 TI - Antibacterial hydroperoxysterols from Xanthosoma robustum. AB - Three new hydroperoxysterols, 24-hydroperoxy-4 alpha, 14 alpha- dimethylcholesta 8,25-dien-3 beta-ol, 25-hydroperoxy-4 alpha, 14 alpha-dimethylcholesta-8,23-dien 3 beta-ol and 25-hydroperoxycycloart-23-en- 3 beta-ol, have been isolated from the aerial parts of Xanthosoma robustum, besides 24-hydroperoxycycloart-25-en-3 beta-ol, 4 alpha, 14 alpha-dimethylcholesta-8,24-dien-3 beta-ol and cycloartenol. Additionally, the two new diols, 4 alpha, 14 alpha-dimethylcholesta-8,25-dien-3 beta, 24-diol and 4 alpha, 14 alpha-dimethylcholesta-8,23-dien-3 beta, 25-diol were obtained from the first two hydroperoxysterols, respectively, by reduction with triphenylphosphine. The structures have been defined by chemical and spectroscopic studies. The four hydroperoxysterols exhibited antibacterial activities against Escherichia coli, Bacillus subtilis and Micrococcus luteus. PMID- 8728720 TI - A benzoic acid glycoside from Geniostoma antherotrichum. AB - Fractionation of an HIV-inhibitory organic extract of Geniostoma antherotrichum afforded a glycoside derivative, which has been characterized as 2-hydroxy-3-O beta-D-glucopyranosyl-benzoic acid (1) on the basis of spectral analyses. The HIV inhibitory activity of the extract was traced to polymeric tannins, while 1 was found to be inactive in the National Cancer Institute's primary anti-HIV screen. PMID- 8728721 TI - A pyrroline glucoside ester and steroidal saponins from Lilium martagon. AB - A new phenylpropanoid ester of a pyrroline derivative and two new steroidal saponins were isolated from the fresh bulbs of Lilium martagon, along with several previously known compounds. The structures of the new compounds were determined by spectroscopic data, hydrolysis, and by comparison with spectral data of known compounds to be (-)-5-hydroxy-3-methyl-3-pyrrolin-2-one 5-O-(6-O-p coumaroyl-beta-D- glucopyranoside), (25S)-spirost-5-ene-3 beta,17 alpha, 27-triol 3-O-[O-beta-D-glucopyranosyl-(1-->2)-O-beta-D-glucopyranosyl- (1-->4)-beta-D glucopyranoside] and (25S)-5 alpha-spirostane-3 beta,17 alpha,27-triol 3-O-[O beta-D-glucopyranosyl-(1-->2)-O-beta-D-glucopyranosyl-(1-->4)-bet a-D- glucopyranoside], respectively. Lilium martagon crosses well with L. hansonii to produce a valuable garden hybrid lily. In this study, the secondary metabolites of L. martagon were revealed to be closely related to those of L. hansonii, giving a good example of the correlation between the secondary metabolites and cross-compatibility. PMID- 8728722 TI - The appraisal of medical scientific evidence. PMID- 8728723 TI - Mandibular-Lingual Repositioning Device--MLRD: preliminary results of 8 patients with obstructive sleep apnea syndrome--OSAS. AB - Dental devices have been employed in the treatment of snoring and obstructive sleep apnea syndrome (OSAS) of mild to moderate degrees. The authors disclose the preliminary results in 8 patients with clinically diagnosed obstructive sleep apnea polisomnographically confirmed and treated with a dental device (Mandibular Lingual Repositioning Device--MLRD) developed by one of the authors (RCB). Cephalometrics was performed before usage of MLRD, Epworth Sleepiness Scale (ESS) tests were conducted before and after the MLRD and a questionnaire that subjectively qualified the Overall Subjective Improvement of the patient (OSI) was filled out. The tests were repeated 4 weeks after the application of the MLRD. The average subjective overall improvement was 73.75% and the Epworth Sleepiness Scale was 13.88 (pre-MLRD) and 6.63 (post-MLRD) representing a significant statistical variation (p = 0.05). The authors discuss probable factors involved in the improvement of the subjective measurements of excessive sleepiness. PMID- 8728724 TI - Abdominal mycobacterial infection in acquired immunodeficiency syndrome patients. AB - The authors studied 12 patients with AIDS and abdominal mycobacteriosis hospitalized in the Hospital Ipiranga (Sao Paulo, Brazil), from June 1989 to January 1992. Diagnosis was confirmed by the histopathological examination of organ specimens collected during laparotomy, which, in most cases, was carried out due to an emergency situation. Observations included perforation of the ileum, seropurulent fluid involved and bloked by viscera, epiploon, and fibrin. Hepatoesplenomegaly was present in all patients and generalized granulomatous peritonitis was observed in more than 50%. A patient died in the immediate post op period, four after an average period of 55 days in the hospital. A patient evolved with stercoral fistula and asked to be discharged. Six patients were discharged after an average hospitalization period of 27 days. The authors stress that in developing regions where tuberculosis incidence is high, a patient with AIDS and a painful and irritative abdominal picture should always lead to the hypothesis of mycobacteriosis. PMID- 8728725 TI - Reduction of liver mass due to malnutrition in rats. Correlation with emaciation of animals and size of organs not inserted in the portal system. AB - We studied the effects of protein-energy malnutrition on the liver morphology of rats as compared to animal emaciation and to reduction in size of the organs not irrigated by splanchnic blood such as kidneys and spleen. The animals were divided into two groups, one of them fed ad libitum rate (N = 10) and the other (N = 14) receiving water but no food for 7 days, and the changes in animal weight, liver, kidney and spleen mass were determined. DNA and the protein/DNA ratio, as well as hepatocyte size, were determined in liver tissue. The liver decreased in mass (27.14%) at a significantly higher proportion (p < 0.05) when compared to body emaciation (19.22%). Similar to the reduction in body weight, the masses of kidneys and spleen were reduced by 18.68% and 24.28%, respectively. The reduction in liver mass occurred due to hypoplasia and atrophy, i.e., a decrease in hepatocyte number and size, respectively. We conclude that there is a preferential consumption of liver protein in protein-energy malnutrition which is suggested to result from the additive action of the effects of overall consumption of organic reserves due to malnutrition proper and to the reduction of the hepatotrophic stimulus. PMID- 8728726 TI - Prognostic factors in 291 patients with pulmonary metastases submitted to thoracotomy. AB - Pulmonary metastases are among the most common sites for the spread of cancer, particularly with mesenchymal tumors. Despite improvements in cancer therapy, the prognosis remains poor, except for the highly selective group of patients who are considered eligible for surgical treatment and those with chemosensitive tumors. The main objective of this study was to evaluate survival results of 291 patients who underwent thoracotomy due to pulmonary metastases between 1953 and 1986. The number of metastases ranged from a minimum of 1 to as many as 30. The type of resection depended on site, size, and number of pulmonary lesions: 154 wedge resections, 49 lobectomies, and 9 pneumectomies. The tumor was not resectable in the remaining 79 patients. Of the prognostic factors analyzed, only type of pulmonary resection (wedge vs. lobectomy vs. pneumectomy vs. unresectable) and disease free interval (DFI) between primary treatment and pulmonary metastases diagnosis (< or = 6 months vs. 7-12 months vs. > 12 months) were selected as independent predictors of the risk of death in multivariate analysis. A reduced model for bone tumors included disease free interval, sex and histology. PMID- 8728727 TI - Nevoid basal-cell syndrome: literature review and case report in a family. AB - The Nevoid Basal-Cell Carcinoma Syndrome (NBCC), or as it is also referred to, basal-cell nevus syndrome or Gorlin-Goltz syndrome, is characterized by multiple early-appearing basal cell carcinomas, keratocytosis of the mandible, and anomalies of the ocular, skeletal reproductive system. We describe four patients in the same family, all of them possessing a large number of skin tumors associated with other typical clinical and X-Ray anomalies of NBCC. The definitive treatment of NBCC has yet to be established, however, early diagnosis is very important as well as the periodical follow-up examination of ten patients, mainly due to the transformations in the skin lesions that may occur. PMID- 8728728 TI - [Nutritional status of school children from different socioeconomic levels]. AB - The aim of this work was to compare the nutritional status of children from low and high socioeconomic levels. Weight, height, mid arm circumference and tricipital skinfold thickness were measured in 1,842 children of low and 2,770 of high socioeconomic status. Mean weight, height, and mid arm muscular circumference were higher in children of high socioeconomic status. Also, growth failure and overweight had a higher frequency among children of low socioeconomic status. Mean weight, height and mid arm circumference were higher in males of both groups. Among children of low socioeconomic status, height/age ratios were lower in men and weight/height ratios were higher in women. These differences were not observed in children of high socioeconomic level. We conclude that adverse environmental conditions, lower physical activity and indigenous ancestors may alter the nutritional status of children of low socioeconomic levels. PMID- 8728729 TI - [Effect of casein on D-xylose absorption assessed by H2 breath test]. AB - Different experimental studies suggest that the presence of food in the alimentary tract, promote small intestinal absorption. The mechanisms involved are not completely understood and might be related to motility changes or to humoral factors. Since studies have shown a decrease of small bowel motility after casein administration, the aim was to analyze the effect of this protein on small intestinal absorption and orocecal transit time. The hydrogen breath test was used to estimate d-xylose absorption. H2 production is dependent on the amount of this carbohydrate reaching the colon, and therefore inversely proportional to d-xylose absorption. Six normal volunteers ingested 25 g d-xylose and 25 g d-xylose + 30 g casein, and alveolar breath samples were obtained at 15 min intervals. RESULTS: by adding casein to d-xylose solution a statistically significant decrease of maximal H2 concentration was observed from a mean of 40 +/- 11 ppm to a mean of 26 +/- 8 ppm. Similarly the area under the curve (which reflects the amount of xylose that was not absorbed) was also significantly decreased from a mean of 3281 +/- 1399 ppm to mean of 1394 +/- 700 ppm of H2. The orocecal transit time was significantly prolonged from 85.5 +/- 40 min to 112 +/- 38 min. Our results suggest that casein increases d-xylose absorption in normal subjects and that this effect might be related to a slower transit time. PMID- 8728730 TI - [Proliferating cell nuclear antigen in cervical cancer and precursor lesions]. AB - The aim of this study was to study the degree of cellular proliferation quantifying the immunohistochemical expression of the proliferating cell nuclear antigen. We assessed paraffin embedded samples of 25 infiltrating epidermoid cervical carcinomas, 76 precursor lesions (34 of low and 42 of high grade), 29 normal and 13 metaplastic epidermoid epithelia. Mean values of proliferating cell nuclear antigen expression were 13.7% in normal epithelia, 15.7% in epidermoid metaplasia, 37.1% in low grade precursor lesions (35.3% in condilomas and 38.8% in mild dysplasia), 48.7% in high grade lesions (47.9% in moderate dysplasias, 50.5% in severe dysplasias and 50% in carcinoma in situ) and 54.7% in infiltrating carcinoma. There were differences in proliferating cell nuclear antigen expression nd mitotic index between low grade precursor lesions and high grade lesions and infiltrating carcinoma. No correlation was observed between mitotic index and proliferating cell nuclear antigen expression. We conclude that infiltrating epidermoid cervical carcinoma and its precursor lesions have a high level of proliferative activity, demonstrated by a high percentage of cells in active phases of the cellular cycle. PMID- 8728731 TI - [In vitro susceptibility of enterohemorrhagic Escherichia coli to 11 antimicrobials. Relationship between antibiotic resistance and toxigenic genotype]. AB - Enterohemorrhagic E coli (EHEC) has been recognized as the main etiologic agent of hemorrhagic colitis and hemolytic uremic syndrome (HUS). The usefulness of antibiotic treatment in patients with EHEC infections is a matter of current debate. Knowledge on EHEC antimicrobial susceptibility patterns in different geographic areas is important for both treatment considerations and for strain characterization. We studied by diffusion disk agar technique the antibiotic susceptibility of 83 EHEC strains obtained from stools of patients with HUS or diarrhea. Eleven antimicrobials were tested (ampicillin, cotrimoxazole, tetracycline, chloramphenicol, furazolidone, gentamycin, amikacin, ciprofloxacin, erythromycin, vancomycin, and metronidazol). Resistant strains by disk diffusion were tested for MIC (mg/ml) by agar dilution. SLT-I and SLT-II were detected with specific biotinylated gene probes. All 83 strains were susceptible to furazolidone, ciprofloxacin, gentamycin and amikacin. Resistance was detected to tetracycline 4%, chloramphenicol 5%, cotrimoxazole 24% and ampicillin 25%. As expected for EHEC strains all were resistant to erythromycin, vancomycin, and metronidazol. Resistant strains were significantly more common in non toxigenic and SLT-I producing strains (p = 0.01). Resistant strains were similarly distributed among patients who had diarrhea only and those who developed HUS (p = 0.3). In Chile, resistant EHEC strains seem to be more common and of different genotypes than those reported in more developed countries. Regional differences of EHEC antibiotic susceptibility patterns indicate a need for continuous monitoring, specially if antibiotics prove to be useful in disease prevention. PMID- 8728732 TI - [Changes in protein repletion markers in undernourished patients receiving enteral nutrition and its relation with energy and nitrogen balance]. AB - AIM: To study the relationship between the changes in protein repletion indicators and calorie and nitrogen balance in undernourished patients receiving enteral nutrition. PATIENTS AND METHODS: Fourteen patients (11 female, age range 55-85 years old) with an initial serum albumin below 3.5 g/dl or serum transferrin below 200 mg/dl, which received enteral nutrition, were studied. On days one and ten of nutritional support, indirect calorimetry, nitrogen balance and serum levels of albumin, transferrin, pre-albumin and total lymphocyte count were measured. RESULTS: Initial resting energy expenditure was 25.1 +/- 5 Kcal/kg/day (115.4 +/- 20% of that predicted by Harris Benedict equations) and urinary urea nitrogen 5.4 +/- 3.7 g/day. Initial and final nitrogen balances were 66.9 +/- 71 and 81.4 +/- 105.9 mg N/kg/day and mean energy balance throughout the study was +9.0 +/- 7.2 Kcal/kg/day. During the ten days of nutritional support, albumin improved significantly in 0.35 +/- 0.43 g/dl (15.2 +/- 19.8%). Transferrin, pre-albumin and total lymphocyte count did not change significantly. The changes in these last parameters did not correlate with nitrogen or calorie balance. CONCLUSIONS: Probably other factors, besides nutritional balances, influence serum protein levels and these may not change despite an adequate nutritional repletion. PMID- 8728733 TI - [Wilson's disease: a review apropos of a clinical experience in 16 patients]. AB - Wilson's Disease is an inherited disorder of copper metabolism. We report 16 patients (6 males) with the disease; 6 had hepatic involvement exclusively, 4 had neurological involvement, 3 had a neurological and hepatic involvement and 3 were asymptomatic. The age onset was 9 years for hepatic and 17 years for neurologic involvement. The mean delay in diagnosis was 14 months. Chronic hepatitis, cirrhosis and fulminant hepatic failure were the clinical forms of liver disease. Patients with neurologic disorders had behavioral disturbances and extrapyramidal manifestations such as dystonia and parkinsonism. Patients had a good response to penicillamine, except 3 that died of liver complications, in whom the treatment was delayed or discontinued. We conclude that this metabolic disease must be suspected in pubertal children and in adults of less than 30 years old with liver disease of unknown origin or behavioral alterations associated to an extrapyramidal syndrome. PMID- 8728734 TI - [Clinical effects of inspiratory muscle training in patients with chronic airflow limitation]. AB - The clinical role of inspiratory muscle training (IMT) in chronic obstructive pulmonary disease (COPD) has not been established, because data on its clinical effect is scarce and controversial. To further investigate these aspects we studied 20 COPD patients (FEV1 37 +/- 3% P) who were randomly and double blindly trained for 30 minutes a day during 10 weeks using a threshold inspiratory trainer with either 30% (group 1) or 10% (group 2) of PIMax as a training load. The training load was crossed after each patient completed 10 weeks of training. Effects were assessed through changes in PIMax, dyspnea through the transition dyspnea index (ITD) and the respiratory effort with Borg's score. Walking capacity was measured with the six minutes walking distance test (6WD) and depression symptoms with Beck's score. Daily life activities were also assessed. Results showed that after 10 weeks of IMT, PIMax increased in both groups (p < 0.05), dyspnea improved in group 1 as compared to group 2 (p < 0.04), 6WD increased significantly in patients of group 1, who also complained of less dyspnea (p < 0.05). Depression scores fell significantly in group 2. Daily activities improved more in group 1. After the crossover patients in group 1 disclosed a significant deterioration in PIMax whereas group 2 disclosed significant improvements in PIMax, dyspnea and 6WD. We conclude that IMT using a threshold device with 30% PIMax is a useful procedure for the treatment of severe COPD patients. PMID- 8728735 TI - [Effect of a 17 beta estradiol gel preparation on hormone levels in menopausal women]. AB - Different delivery forms of supplemental estrogens may differ in their hormonal effects. The aim of this study was to assess the effects of a daily dose of 2.5 g of 17 beta estradiol transdermal gel, given during four weeks, on hormone levels of six postmenopausal women. At the fourth week we observed a significant increase in estradiol and a decrease in FSH levels. Estrone levels also increased but the estradiol/estrone ratio was maintained in values over 1. No changes in SHBG or IGF1 levels were observed. Two patients that used the gel in the abdominal skin achieved lower estradiol levels (below 60 pg/ml). We conclude that the gel increased serum estradiol levels over 60 pg/ml in four of six women, that there is a big individual variability and the application zone could influence the serum estradiol levels achieved. PMID- 8728736 TI - [Evaluation of bone mass in hip fractures measuring lumbar spine and contralateral hip with bone densitometry]. AB - Hip fracture is frequent in postmenopausal women with osteoporosis. The aim of this work was to assess bone mass in women with hip fracture and compare it with that of normal women. Bone densities of lumbar spine (considering areas with and without spondylosis), femoral neck, greater trochanter, and unfractured hip Ward's triangle were measured with a double beam isotopic densitometer. Thirty one women aged 58-95 years old were studied and compared with normal women studied at the same laboratory. Bone densities in fractured and normal women were 0.82 +/- 0.16 and 0.85 +/- 0.05 g/cm2 in lumbar spine respectively (NS), 0.74 +/- 0.15 and 0.85 +/- 0.05 g/cm2 in lumbar spine without spondylosis respectively (p < 0.001), 0.60 +/- 0.11 and 0.65 +/- 0.08 g/cm2 in femoral neck respectively (NS), 0.49 +/- 0.09 and 0.57 +/- 0.05 g/cm2 in greater trochanter respectively (p < 0.001) and 0.48 +/- 0.12 and 0.52 +/- 0.09 g/cm2 in Ward's triangle respectively (NS). It is concluded that the larger differences in bone density between women with and without hip fracture are observed in the greater trochanter. PMID- 8728738 TI - [Inflammatory lung pseudotumor]. AB - We report a 63 years old man presenting with a history of persistent cough and blood streaked sputum. Chest X rays and CAT scans showed a tumoral lesion between the superior and inferior right lobes. The tumor was resected and its pathological examination revealed an inflammatory pseudotumor. It is concluded that this type of tumors must be born in mind in patients with nodular lesions of the lung. PMID- 8728737 TI - [False high levels glycosylated hemoglobin in 2 diabetic patients with persistence of fetal hemoglobin]. AB - Two diabetic patients with unusual high levels of glycosylated hemoglobin measured by ion exchange chromatography are described. Further studies revealed a persistence of fetal hemoglobin in both cases. This condition produces falsely high levels of glycosylated hemoglobin, when ion exchange chromatography is used. These cases may be overtreated with risk of hypoglycemia. Patients with inappropriate levels of glycosylated hemoglobin should be investigated for hemoglobinopathies. PMID- 8728739 TI - [Advances in the treatment of tachiarrhythmias: role of nonpharmacological methods]. AB - Nonpharmacological methods are a novel therapeutic option for tachiarrhythmias. Transcatheter ablation or surgery can cure many arrhythmias, avoiding the collateral effects of antiarrhythmic drugs. Likewise, implantable defibrillators, have changed life expectancy of patients with high risk arrhythmias or sudden death survivors. However, the high cost and sophistication of these methods, preclude their widespread use, thus limiting the number of patients that can be benefited. This article reviews the main nonpharmacological techniques for treatment of arrhythmias, their results and complications. PMID- 8728740 TI - [Graduate education financing. An indispensable review in the modernization of health education in universities]. AB - Due to the social and economical changes in the Chilean university and health systems in the last decades, the traditional financing model of the Faculty of Medicine and postgraduate training programs became obsolete. The progressive reduction of the fiscal budget has forced the search of alternate financing sources. The payment of specialization is, at the present moment, an obliged, inappropriate and without compensation subsidy of the universities to the public and private health system, that profit of training professionals and to the specialists that acquire skills that will increase their earnings. Equable financing systems should be established in which the direct beneficiaries, institutions or individuals, would pay the expenses. A national scholarship fund or deferred credits seem reasonable alternatives that should be promptly established to allow the survival of university postgraduate training programs. PMID- 8728741 TI - [Development and validation of a survey for evaluation of the satisfaction of patients with medical consultation in primary care clinics]. AB - The aim of this work was to devise, and assess the reliability, of an instrument to measure satisfaction with medical consultation of primary care urban patients at public outpatient clinics. A model was elaborated, based on data collected from the literature and from focal groups of patients and physicians, and an enquiry was developed and applied at four outpatient clinics. Fifty enquiries were obtained and analyzed to discard questions of difficult understanding. Next, a second self administered questionnaire and with fewer questions was devised. Its reliability was assessed in 53 enquiries, obtaining a Cronbach's a of 0.904. Questions that decreased consistency were discarded, finally obtaining a self administered instrument with 22 questions. This enquiry was used in 174 patients. Results were subjected to factorial analysis with varimax rotation, which separated three factors that explain 64% of the variance. It is concluded that a valid and reliable instrument was obtained. PMID- 8728742 TI - [During the centenary of Pasteur's death (1822-1895)]. AB - Pasteur had annoying opponents at the Medical Academy of Paris, probably because he was not a physician. The medical aspect of his creation reached highest altitude when he vaccinated the boy Jose Meister against rabies. By that time, he was crippled by a hemiplegia suffered when he was 46 years old. Studying tartric acid isomer, Pasteur discovered molecular asymmetry. He hypothesized that life operated asymmetrically, when he discovered the selectivity of Penicillium glaucum fungus action on paratartrates. He discovered anaerobic life and discarded the theory of spontaneous generation. At the age of 70, he received the gratitude of France and the whole mankind, through President Carnot. On that occasion he said to youngsters "Live in the calm peace of laboratories and libraries. Ask yourselves: What have I done for my instruction?, What have I done for my country?, until the moment that you reach the immense happiness of thinking that you have contributed to mankind's progress and welfare." PMID- 8728743 TI - [Influence of body weight in pregnancy and the perinatal results]. AB - Between 1988-1922, data of the nutritional status of pregnant women seen in the Santiago Metropolitan Health Service were analyzed. Underweight (22.2%), normal weight (47.2%), overweight (19.7%) and Obese (15.4%). Four thousand five hundred fifty five pregnant women were studied. Underweight 1136, normal weight 1219, overweight 1100 and obese 1100. Underweight was significantly more frequent in the patients less than 20 years old while overweight and obese was significantly more frequent in the patients over 30 years old. Hypertension (2.6%) was the only significant morbidity factor in the obese group. The overweight and obese groups had earlier menarche, while the obese group had shorter periods. The obese group were associated most frequently with higher parity (75.1%), stillbirth (4.6%), spontaneous abortion (19.5%), induced abortion (3.1%) and high obstetric risk (33.2%). In the normogram used, the underweight patients are abnormally represented at the start of pregnancy. The obese group gained less weight proportionally during pregnancy (overweight and obese 42.8%, underweight and normal 34.7%). The obese group presented more frequently with hypertension (20.4%) and diabetes (0.7%), while the obstetric complications occurred more frequently in the underweight (6.3%). The underwent group had more anemia (45.4%) and premature labor (12.3%). Cesarean section was performed more frequently in the obese group (33.1% versus 21.3% of all the other groups combined. The neonatal birthweight was in direct proportion to the maternal weight, measured by various methods. It is worth noting the importance of microelements in the milk ingestion of the pregnant patients and the influence on their weight. PMID- 8728744 TI - [Correlation between plasma progesterone and ultrasonographic detection of corpus luteum in ovulatory cycles]. AB - The ultrasound detection of corpus luteum in the midluteal phase of ovulatory cycles was associated with significantly higher levels of plasma progesterone, opposed to those patients where a corpus luteum could not be identified. The hormone level was directly proportional no the number of corpora lutea seen. However, only 33.3% o patients with no corpus luteum visible were found to have suboptimal plasma progesterone levels (< 10 ng/ml). PMID- 8728745 TI - [Unexplained increase in maternal plasma in alpha-fetoproteins in the second trimester of pregnancy: perinatal results]. AB - To determine the relation between adverse perinatal events and an unexplained mid trimester elevation in maternal serum alpha-fetoprotein (MSAFP) in an unselected population, 16,093 women with a singleton pregnancy delivered between January 1985 and November 1991, at the Queen Mother's Hospital, Glasgow, Scotland, were studied. The elevated MSAFP group (n = 606) was associated with an increased risk of preterm delivery (odds ratios/95% confidence intervals) (3.7/2.6-5.2), small or gestational age (4.5/3.3-6.1), intrauterine death (3.9/1.7-9.4), and placental abruption (3.2/1.5-6.7). The risks increased with MSAP concentration. An unexplained mid-trimester elevation of MSAFP is associated with increased risk of adverse perinatal events, thus acting as a non-specific marker o a "high risk pregnancy". Alterations to current antenatal management in this group may improve prognosis. PMID- 8728746 TI - [Cervical pregnancy. 3 clinical cases]. AB - Three cases of cervical pregnancy are diagnosed by transvaginal ultrasonography. The authors emphasized the opportune medical treatment, avoiding surgical risk. PMID- 8728747 TI - [Ovarian function determination in pre and postsurgical sterilization]. AB - The menstrual characteristics, FSH, LH, estradiol and progesterone levels of 21 women were measured before and one month after sterilization by tubal ligation. No statistical differences were found. PMID- 8728748 TI - [Results in the treatment of 82 cases of endometrial cancer. Surgical staging vs clinical staging]. AB - Eighty two cases of endometrial carcinoma were treated from 1971 to 1994. From them twelve patients were clinically staged and 70 were staged according to 1989 FIGO classification. Thirty five patients received complementary total radiation therapy to the abdomen, pelvis and intravaginal depending on the depth of miometrial invasion, positivity of peritoneal cytology or extrauterine involvement. Thirty two patients presented with carcinoma localized to the endometrial layer received no complementary treatment. Three patients were under exploratory laparotomy with surgical non curative resection due to massive abdominal tumor invasion. The follow up revealed 59 patients free of disease (77.6%). The relapse date was 22.3%. The relapse in the clinically staged patients (n = 12) was 41.6%. The mortality rate was 15.8%. PMID- 8728749 TI - [Predictive value of endometrial pattern and thickness in the result of in vitro fertilization and embryo transfer]. AB - In order to assess the relationship between echographic endometrial thickness and pattern and the probability of clinical pregnancy and delivery we analyzed retrospectively 51 cycles of IFV-ET. Patients were classified in three categories of endometrial thickness: < 9 mm, 9.1-11 mm and > 11 mm and in two types of endometrial pattern: type 1 (non triple line) or type 2 (triple line). The average endometrial thickness of patients who got pregnant and those who did not was 11.8 +/- 1.8 mm and 10.6 +/- 1.7 mm respectively (p = NS). There were no clinical pregnancies in patients with endometrial thickness < 9 mm (p < 0.01). The average endometrial thickness in patients with endometrial pattern type 1 and type 2 was 11.1 +/- 1.2 mm and 10.9 +/- 1.9 mm respectively (p = NS). Patients who presented type 2 endometrial pattern (n = 39) produced eleven clinical pregnancies (28.2%) and those with type 1 (n = 2) only one clinical pregnancy (8.3%), which ended in spontaneous abortion (p < 0.01). When both variables, thickness and pattern, were considered together, patients with endometrial pattern type 2 and thickness > 11 mm had a probability of clinical pregnancy and of delivery of 43.8% and 25%, respectively, who is statistically significant (p < 0.03). We conclude that endometrial thickness and pattern, taken together, offer a valuable prognostic value for the outcome of an IVF-ET cycle. PMID- 8728750 TI - [Acute fulminating hepatitis non-A non-B in pregnancy: a clinical case of probable sporadic hepatitis caused by E virus]. AB - The extreme severity of hepatitis E in the pregnant patient results a high maternal and fetal mortality in fulminating acute hepatitis. We present a clinical case of a 19 years old primigesta, 29 weeks pregnant, diagnosted as suffering from Acute Fulminating Hepatitis non A non B, probably E. The fetus died and liver transplant was contemplated for the patient due to the severity of disease. Liver transplant could not be carried out for lack of a suitable donor. The patient died 16 days after admission. The pathology report showed massive hepatic necrosis and terminal shock. PMID- 8728751 TI - [Von Recklinghausen's disease (neurofibromatosis) and pregnancy: apropos of a clinical case]. AB - We present a case of von Recklinghausen's disease in a patient whose pregnancy resulted in a stillbirth in the third trimester, probably due to placental insuficiency. Although neurofibromatosis is one off the most frequently occurring genetic diseases, its diagnosis is rarely made. PMID- 8728752 TI - [Cystic hamartoma: prenatal diagnosis and follow-up of newborn infants]. AB - A case of fetal hamartoma cyst diagnosed prenatally was successfully removed surgically after birth. PMID- 8728753 TI - [Pregnancy and the use of pacemakers]. AB - We present a 19 year old patient with complete atrioventricular block, who had a permanent demand type pacemaker-implanted. Her pregnancy and labor were uneventful. There were no maternal nor neonatal complications. Since the association of pregnancy and pacemaker use is rare, we present our experience and management of the case. PMID- 8728754 TI - [Exceptional variety of endometrial cancer]. AB - We present a clinical case and histology of rare endometrial neoplasm: Giant cell carcinoma of the endometrium. PMID- 8728755 TI - [Risk factors for Ureaplasma diversum infections in cattle of a tropical environment]. AB - A case-control study, determined the influence of breed, age and number of deliveries as risk factors for Ureaplasma genital infections in Costa Rica dairy cattle. The animals with none or one delivery had a risk of infection 2.99 times higher than those with several parturition, regardless of breed. The risk was 1.95 times higher in Jersey than in Holstein, and decreased progressively until three deliveries. In cows with four deliveries there was a significant increase in the amount of animals infected and also a significant difference in the rate of infection between Holstein (27%) and Jersey (64%). Ureaplasma isolation was rare in cows with more than six deliveries. PMID- 8728756 TI - Age influence in the natural resistance of white rat and mice to the protozoan Toxoplasma gondii. AB - Groups of 1, 5, 10, 15 and 30 day old rats (Sprague Dawley) and Wistar mice were infected per os with different inocula of Toxoplasma gondii oocysts from an avirulent strain. Infection pattern was followed by determination of survival time, tissue lesions, parasite presence and serology. Although all the animals survived the total experimental period (30 day), there were important differences between both rodents regarding to Toxoplasma natural resistance. Brain cysts, as well as tachyzoites in other organs were regularly present in mice but not in rats. On the other hand, most of the organs from mice presented characteristic T. gondii lesions, while in rats a normal tissues were observed, with the exception of an acute reaction found in the brain of a 15d old animal. PMID- 8728757 TI - [Karyotyping of fetal cells in the prenatal diagnosis in Costa Rica]. AB - The results of 182 genetic amniocenteses between 14 and 37 weeks gestation, from 1986 to 1992, and of two cordocenteses in 1992, are reported. There were two main reasons for referral: maternal age 35 years and older and abnormal ultrasound assessment. Fetal cells were closed cultured and mass harvested. In 3.7% of cases fetal chromosomes were defective. Turn around time was about three weeks up to and including 1991 and two weeks in 1992, culture failure rate was 7% that year. No cytogenetic misdiagnosis and no complication or sequelae related to the amniocenteses were detected. We conclude this is a safe and reliable procedure to obtain fetal karyotypes and to improve obstetric management of high-risk pregnancies. PMID- 8728758 TI - [Fatty acids contained in 4 pejibaye palm species, Bactris gasipaes (Palmae)]. AB - Cooked and uncooked samples from four pejibaye palm races were analyzed to determine the moisture content, ether extract and the content of six fatty acids (C16:0 to C18:3). There was an increase in moisture and a decrease in ether extract (p < 0.05) in the cooked samples in comparison with the uncooked ones. No significant differences were found in fatty acid content between cooked and uncooked samples, but there were differences (p < 0.05) among races concerning the content of four fatty acids. Pejibaye fat is mainly mono-unsaturated (45.6%) and has a low poly-unsaturated to saturated fatty acid ratio (0.5). The fatty acid profile of uncooked pejibaye samples was: oleic acid, 32.6 to 47.8%; palmitic acid, 30.5 to 40.3%; linoleic acid, 11.2 to 21.1%; palmitoleic acid, 5.7 to 7.1%; linoleic acid, 1.5 to 5.5%; and stearic acid, 1.7 to 2.4%. PMID- 8728759 TI - [Accumulation and depuration of aldrin in the oyster Crassostrea rhizophorae, of the Cienaga Grande de Santa Marta (Colombian Caribbean)]. AB - Laboratory experiments were conducted at different salinities (35, 26 and 17/1000) to determine accumulation and depuration rates of the organochlorine compound aldrin by the oyster Crassostrea rhizophorae, Cienaga Grande de Santa Marta (Colombian Caribbean). Residual levels were detected in tissues after 24 hours of exposure, showing increases directly related to the exposure concentration. For all the exposure concentrations the highest retention values were measured at 26/1000 salinities, followed by 17/1000, and the lowest values at the highest salinity. The highest bioconcentration values were 96115.5 (at 26/1000), 37938.9 (at 17/1000), and 22605.1, (35/1000) times the environmental concentration. The depuration process varied across salinity levels. At 26/1000 there was a clear decrease in the residual levels during the first five days, eliminating in this period the same amount as oysters kept at 17/1000 for 20 days and surpassing the amount eliminated by those kept at 35/1000 for 20 days. A rapid decrease in the amount of residues is followed by a slow elimination of the aldrin remaining in tissues. PMID- 8728760 TI - [Biometry of Modiolus capax (Bivalvia: Mytilidae) in Playa Ocotal, Guanacaste, Costa Rica]. AB - 155 specimens of mussels (Modiolus capax) were gathered during March 1993 in a subtidal bed, located in northern Ocotal, Guanacaste, Costa Rica. For each individual in the random sample, shell length, breadth and height were measured and total weight, shell weight empty and meat weight were controlled. Analysis of data determined relationships of length to weight, length to breadth and length to height of Lt = 1.31977 Pt + 29.05478, Lt = 0.36605 D + 3.98744 and Lt = 0.50434 A + 2.73786. The mean Conditional Index for the population was 39 +/- 0.07. PMID- 8728761 TI - [Nutrition of juvenile prawn Macrobrachium carcinus (Crustacea: Decapoda) with diets of vegetable and marine residues]. AB - Juvenile prawn Macrobrachium carcinus were fed two different diets: restaurant by products (diet I) and fish and vegetable market by-products (diet II). These diets were evaluated by proximal analysis, assimilation efficiency and the factor conversion rate (FCR). Diet I registered a higher efficiency, but there was no difference in the growth rate. The growth mean (G. L.) for three months was 0.254 +/- 0.13 cm (diet I) and 0.191 +/- 0.1 cm (diet II). The conversion rate was good for both, suggesting that 6 to 7 kg of food are needed to obtain 1 kg of prawn. Survival was 76% and 100% for diets I and II, respectively. PMID- 8728762 TI - [Determination of some hematological aspects of Oncorhynchus mykiss (Salmonidae), in Cundinamarca, Columbia]. AB - Thirty-five normal, adult specimens of the rainbow trout Oncorhynchus mykiss (Richardson, 1836) cultured in the Neusa Fishculture Station (73 degrees 38'28" W; 5 degrees 8'30" N) were studied to establish normal blood values; the hematopoietic tissues (pronefros, spleen and thymus), and other organs such as the stomach were also structurally studied. The rainbow trout collected in the second half of 1990 and the beginning of the first half of 1991 had 1,415,000 erythrocytes and 22,000 leukocytes per mm3 of blood. Their hemoglobin concentration was 6.99 g/100ml. They had three types of circulating leukocytes: lymphorytes 65%, polimorphonuclears 24% and monocytes 11%. The pronephros is the major hematopoietic organ in the rainbow trout. It produces mainly the polimorphonuclears; the spleen and the thymus are linfopoietics. The study of the hemocytometric values in the peripheral blood frotis and the structure of the hematopoietic organs of the rainbow trout is very useful, accessible and inexpensive. It produces data which yield information about physiological and pathological conditions. PMID- 8728763 TI - The effect of food and frozen storage on the nutrient composition of some African fishes. AB - Specimens of Tilapia zillii, Clarias lazera, Channa obscura, Synodontis schali, and Scomberomus tritor were collected between January and December 1990, covering the dry and rainy seasons. According to stomach analysis they were classified as either herbivorous, carnivorous, omnivorous, plankton consumers or invertebrate feeders. The crude protein, lipid and moisture contents were determined both for pre- and post frozen storage, with the exception of S. tritor (only post storage data). The percentage of total lipid and protein decreased significantly after each succeeding frozen storage, and the moisture content shows a trend similar to other parameters in all the species. Fresh fish is of the highest nutritional value. The feeding habit had some relationship with the muscle protein, fat and moisture content. PMID- 8728764 TI - Oxybelis wilsoni, a new species of vine snake from Isla de Roatan, Honduras (Serpentes: Colubridae). AB - A new species of Oxybelis is described from Isla de Roatan, Honduras. It differs from all species of Oxybelis in its tan to golden yellow coloration, in having a higher number of subcaudals as well as in other details of color and scutellation. The new species is closely related to O. fulgidus and is thought to be derived from a fulgidus-like ancestor (i.e., is the sister species to fulgidus) that has been isolated in the area of its present range probably since the very late Pliocene. The presence of a yellow population of Oxybelis on Isla de Roatan, Honduras, has long been known (Keiser 1969). The status of this insular population has remained uncertain pending the acquisition of additional material (Keiser 1969, Wilson and Meyer 1985). We recently visited Isla de Roatan a number of times and obtained a large series of additional specimens. After examining this new material, and the previously known specimens from this island, we believe that this population represents an undescribed taxon, which should be known as. PMID- 8728765 TI - Reproduction and feeding behavior of Oxybelis wilsoni, a new species of vine snake (Serpentes: Colubridae). PMID- 8728766 TI - A case of human bite by the pelagic sea snake, Pelamis platurus (Serpentes: Hydrophiidae). PMID- 8728767 TI - [Quantitative Tc 99m - DISIDA hepatobiliary scintigraphy in normal and cirrhotic alcoholic subjects]. AB - Quantitative hepatobiliary scintigraphy (Q.H.S.), with 99m Tc-DISIDA was performed on 15 control subjects and 32 alcoholic cirrhotic patients (A.C.). We used a dynamic planar scintigraphy (30 seg/frame, up to 45 min) technique following injection intravenously of 99m Tc-DISIDA. Time/activity curves were obtained from the right upper lobe of the liver and the: 1) slope uptake, 2) half time (T 1/2 min) uptake, 3) excretion half-time (T1/2 min), were measured from the curve. The A.C. were divided in two groups, IIA (n = 32) and IIB (n = 6) if the excretory curve show negative slope or not respectively. RESULTS: The mean value (+/- 1 D.S. 95% coinfidence interval) of the slope uptake of the A.C. IIB (1.2 +/- 0.40) was significantly slower than a.C. IIA (2.8 +/- 0.39) and control (4.5 +/- 1.17, p = 0.0001 respectively). The difference also was significantly when the mean of A.C. IIA was compared to control (p = 0.007). The mean of T1/2 uptake of A.C.IIB (62.2 +/- 22.2) was significantly longer than A.C. IIA (28.4 +/ 4.4 p = 0.011) and control (17.9 +/- 3.87, p = 0.003). The mean T1/2 excretory of the A.C. IIA (90.0 +/- 17.8) was also significant delayed compared to the mean of normal control (35.6 +/- 7.6 p = 0.001). In the A.C. IIB the excretion plateau curve was associated with visualization of the gallbladder and bowel activity suggesting that the excretion of the IDA preferentially came from the left hepatic lobe. We conclude that alcoholic cirrhotic patients have impaired the mechanism related with the uptake/excretion transport of organic anion, and suggest that noninvasive Q.H.S. with 99m Tc-DISIDA, can be a useful clinical technique to be used for the quantification of hepatic function in cirrhotic alcoholic patients. PMID- 8728768 TI - [Immunoperoxidase in the studies of immune thrombocytopenias]. AB - The usefulness of different techniques to measure platelet bound IgG has been reviewed by George. We present here the results obtained with a technique designed to measure membrane bound IgG employing an anti-human IgG labeled with peroxidase and using O-dianisidine-H2O2 to reveal the enzymatic activity. We studied 152 patients with chronic autoimmune thrombocytopenic (ATP) including 120 adults and 32 children (age below 15 years old), diagnosed by exclusion of diseases that may be associated with thrombocytopenic purpura of either immune or nonimmune mechanisms. Besides, 79 patients with thrombocytopenia related to other diseases were also evaluated. The normal values in 215 controls were 188 +/- 4 IgG molec/platelet (mean +/- SE), while in the whole population of chronic ATP the results were 4714 +/- 344, p < 0.001. In pediatric cases the results had a tendency to values higher than in adults. A negative correlation was found between the number of platelets and the amount of bound IgG, r = 0.41 p < 0.001. IgG bound platelets were also increased in treated patients at relapse. The percent of normal IgG bound platelet was 4.5% in patients with a platelet count below 50,000/microliters and 39% in those with normal platelet number. Patients with secondary thrombocytopenia had elevated IgG/platelet while the values were normal in patients with thrombocytopenia of unknown etiology. We conclude that the immunoperoxidase technique is useful to establish the immunologic nature of thrombocytopenia. PMID- 8728769 TI - [Inactivation of the myocardial lipoamide dehydrogenase by catecholamines. Prevention by captopril and other thiol compounds]. AB - Inactivation of lipoamide dehydrogenase (LipDH) by the Cu(II)/H2O2 Fenton system (SF-Cu(II): (5.0 microM Cu(II), 3.0 mM H2O2) was enhanced by catecholamines (CAs), namely, epinephrine, levoDOPA (DOPA), DOPAMINE, 6-hydroxyDOPAMINE (OH DOPAMINE) and related compounds (DOPAC, CATECHOL, etc.). After 5 min incubation with the Cu(II)/H2O2/CA system (0.4 mM CA), the enzyme activity decayed as indicated by the following percentage values (mean +/- S.D.; in parenthesis, number of determinations): SF-Cu(II) alone, 43 +/- 10 (18); SF-Cu(II) + epinephrine, 80 +/- 9 (5); SF-Cu(II) + DOPA, 78 +/- 2 (4); SF + Cu(II) + DOPAMINE, 88 +/- 7 (5); SF-Cu(II) + OH-DOPAMINE 87 +/- 6 (7); SF-Cu(II) +/- DOPAC, 88 +/- 3 (6); SF-Cu(II) + catechol, 85 +/- 6 (5). In all cases P < 0.05, with respect to the SF-Cu(II) control sample. CAs effect was concentration dependent and at the 0-100 microM concentration range, it varied with the CA structure. Above the 100 microM concentration, CAs were equally effective and produced 90-100% enzyme, inactivation (Figure 2). In the absence of oxy-radical generation, the enzyme specific activity (mean +/- S.D.) was 149 +/- 10 (24) mumol NADH/min/mg protein. Assay of HO. production by the Cu(II)/H2O2/CA system in the presence of deoxyribose (TBA assay) yielded values much greater than those obtained omitting CA. Hydroxyl radical production depended on the presence of Cu(II) and H2O2 and significant H. values were obtained with OH-DOPAMINE, DOPAC, epinephrine, DOPAMINE, DOPA and catecol supplemented systems (Table 2). LipDH (1.0 microM) inhibited 50-80% deoxyribose oxidation, the inhibition depending on the CA structure (Table 2). Native catalase (20 micrograms/ml) and bovine serum albumin (40 micrograms/ml) effectively prevented LipDH inactivation by the Cu(II)/H2O2/CA system; denaturated catalase, SOD, 0.3 M mannitol, 6.0 mM ethanol and 0.2 M benzoate were less effective or did not protect LipDH (Table 3). Incubation of CAs with the Cu(II)/H2O2 system produced a time and Cu(II) dependent destruction of CAs, the corresponding o-quinone, production as illustrated with epinephrine (figures 6 and 7), as illustrated with epinephrine and DOPAMINE (Table 4). These results support LipDH inactivation by (a) reduction of Cu(II) to Cu(I) by CAs followed by Cu-catalyzed production of HO. from H2O2; (b) CA oxidation followed by the corresponding o-quinone interaction with LipDH. CAPTOPRIL, N-acetylcysteine, mercaptopropionylglycine and penicillamine prevented to various degree LipDH inactivation by the Cu(II)/H2O2/CA systems (Table 1). The former was the most effective and 0.4 mM CAPTOPRIL prevented about 95-100% the effect of Cu(II)/H2O2/CA systems supplemented with epinephrine, DOPAMINE and OH DOPAMINE (Figures 3 and Table 1). LipDH increased and CAPTOPRIL inhibited epinephrine oxidation by Cu(II)/H2O2 (Figures 4 and 5). Since un-physiological concentrations of CAs and Cu(II) may be released in the myocardium after ischemia reperfusion, the summarized observations may contribute to explain myocardial damage in that condition. PMID- 8728770 TI - [Effects of estrogen therapy on bone mass in postmenopausal women with osteopenia]. AB - Estrogen treatment prevents early postmenopausal bone loss. The aim of this study was to evaluate the effects of estrogen-progestogen therapy on bone mass in elderly osteopenic postmenopausal women. Fifteen women with a mean age of 58 +/- 6 (mean +/- SD) years and 12 +/- 7 (5-31) years from menopause were evaluated. Bone mineral density (BMD) was assessed by dual X-ray absoptiometry (DXA) with 1.7% variation coefficient at lumbar spine (L2-4) and 1.9% at femur neck. Measurements were done at both sites before and after a 12 month treatment. At the beginning of the study lumbar spine BMD (LS BMD) was low: < 0.9 grs/cm2; z score: -1.4 +/- 0.17 (mean +/- SEM). Treatment consisted in transdermal 17 beta estradiol (50 micrograms/day) (n = 10) or an equivalent natural estrogen oral dose (n = 5). Variable doses of medroxiprogesterone acetate were added on an individualized basis to women with an intact uterus (n = 12). Calcium intake was increased up to a median of 1200 mg/day (800-1600). After a one year treatment LS BMD was increased by 8.4 +/- 1.1% (mean +/- SEM) (95% CI: 6-10.8), from 0.748 +/- 0.02 to 0.810 +/- 0.02 gr/cm2 (p < 0.0001). A less marked gain in femur neck bone mineral density (FN BMD) was also noticed: 3.9 +/- 1.5% (95% CI: 0.6-7.2); 0.671 +/- 0.02 vs 0.697 +/- 0.02 gr/cm2 (p < 0.05). Patients treated with transdermal and oral routes showed similar results. Percentage variations in LS BMD and FN BMD were positively correlated (r: 0.53; p < 0.05). Six patients were treated for 2 years; LS BMD continued to rise, the additional gain being 5.1 +/- 2.2% (p < 0.05), while a non significant increase in FN BMD was observed (7.5 +/- 3.5%; p = 0.06). In the early postmenopausal period, hormonal replacement therapy (HRT) produces either a stabilization or a slight increase (2-4%) in BMD. In contrast, a significant augmentation of bone mass (especially at the spine) seems to occur in osteopenic women when THR is administered in the late postmenopausal period. This suggests that HRT could be used for the prevention as well as for the treatment of postmenopausal osteoporosis. Further studies should be done to evaluate whether HRT reduces the incidence of osteoporotic fractures in elderly osteopenic women. PMID- 8728771 TI - [Immunohistochemical analysis of p53 and c-erbB-2 in breast cancer]. AB - We have studied by immunohistochemistry the nuclear expression of p53 and the finding of oncoprotein c-erbB-2 in a series of 85 breast carcinomas. The tissue was fixed in buffered formalin and embedded in paraffin. The primary antisera were obtained from Biogenex (USA), Bp53-12, monoclonal, used in a dilution of 1:100 for p53 and from Triton Diagnostics (USA), pAB-1, polyclonal, used in a dilution of 1:200 for c-erbB2. The detection system was the Vectastain Elite kit obtained from Vector Laboratories (USA). The results were compared with several morphological features of the tumors such as size and type of the tumor, extension of the intraductal component, nuclear grade, axillary lymph node metastasis and estrogen receptor data as well as with total and disease free survival. The oncoprotein p53 was detected in the nuclei in 25 (29.4%) of our cases (Fig 1). Its presence was correlated with tumor size (p < 0.01), high nuclear grade (p < 0.0001) and estrogen receptor negative tumors (p < 0.0001). There was an inverse relationship between p53 expression and 5 year disease free survival (p < 0.005). In 21 (24.7%) of the cases we found amplification of c-erbB 2. The staining pattern was membranous (Fig. 2). It was correlated significantly with the ductal type of invasive carcinoma (p < 0.05), an associated extensive intraductal component (p < 0.001), estrogen receptor negative tumors (p < 0.0001) and shortening of disease free survival in the patients with positive axillary lymph nodes (p < 0.005). In 9 cases we found staining for both markers without statistically significant relationship with the other features studied. We conclude that the presence of these genetic alterations demonstrated by immunohistochemistry were, in our series, unfavourable prognostic factors in invasive breast cancer. PMID- 8728772 TI - [Diabetes mellitus and hypertension, clinical and epidemiological aspects in the population of La Plata]. AB - The purpose of this study was to determine socio-demographic characteristics, habits, most frequent morbid associations and degree of compliance with the control and treatment of their illness in a population of diabetic and hypertense patients of the La Plata area. A representative sample (890 people) was selected through a home survey (413 housing units). The results obtained show that diabetic and hypertense people a) are in average older than the general population and that the percentage of sedentary habits among them is also higher; b) show multiple typical symptoms of the illness but do not identify them as such and consequently diagnosis is frequently haphazardous; c) have a higher frequency of association with other risk factors, intercurrencies and hospitalization; d) are treated mainly by giving priority to drugs over changes in their detrimental habits; e) tend to ignore those indications that prescribe a change in their habits and f) control their illness at an inadequate periodicity. Consequently, it would be advisable to emphasize the incorporation of education strategies into the treatment of these patients in order to give more importance to preventive and health promoting actions. Education programmes should include not only patients and their families but also members of the health team and the community in general. PMID- 8728773 TI - A brain soluble fraction inhibits synaptosomal membrane but not astrocyte ATPase activity. AB - Since a brain soluble fraction (peak II) is known to be able to inhibit synaptosomal membrane Na+, K(+)-ATPase activity, here we attempted to compare its effect on cellular and subcellular brain components such as synaptosomal and astrocytic membranes, as well as mitochondrial preparations. The difference between total and Mg(2+)-ATPase activity was noteworthy in synaptosomal membranes but proved unremarkable in astrocytic and mitochondrial preparations. Peak II highly inhibited total ATPase in synaptosomal membranes but failed to modify enzyme activity in astrocytic and mitochondrial preparations. Findings suggest cellular and subcellular specificity of peak II on brain ATPase activity. PMID- 8728774 TI - [Meningitis due to Enterococcus faecalis]. AB - Enterococcus faecalis meningitis is an infrequent entity that accounts for less than 1% of all suppurative meningitis in the adult. Usually, this infection affects patients with compromised host defenses or those who have congenital or acquired CNS lesions mainly as intrahospitalary infections. An 85 year old woman from our community (Tandil county) without any predisposed condition, was admitted in the hospital in an unconscious state (grade 3 Glasgow's index), meningeal signs and purulent CSF, from which E. faecalis was isolated. The patient was treated with IV Ampicillin and Gentamycin (17 days), intrathecal Gentamycin (4 days) and IV dexametasona (6 days). The clinical and bacteriological remission was achieved, without any sequel or relapse during 2 years follow up. PMID- 8728775 TI - [Senile dementia, hepatopathy and hydrocephalus]. PMID- 8728776 TI - [Ion channels in non excitable cells]. AB - Several distinct types of voltage-gated and second-messenger-operated K+, Ca2+, Na+ and Cl- channels exist in electrically non excitable cells such as those of the hematopoietic lineage. In these cells ion channels mediate cellular functions involving intracellular biochemical responses, rather than rapid electrical signaling. The presence of the channels is required for several basic functions, such as activation, secretion of lymphokines, mitogenesis, the regulation of cell volume and the mechanisms of resistance to chemotherapeutic agents. Here IN we review the patch-clamp method for studying many characteristics of these ionic channels, particularly in blood cells. PMID- 8728777 TI - [Last advances in Fragile X chromosome syndrome]. AB - The Fragile X syndrome is one of the most frequent forms of mental retardation. The responsible mutation is an unstable repetitive sequence. Since the mutation's discovery, the knowledge about the gene, its protein, function, expression, laboratory detection, phenotype-genotype relationship and risk of expansion, has enormously increased. This work pretends to review the recent advances in this syndrome in all its aspects. PMID- 8728778 TI - [Retrovirus and cancer]. PMID- 8728779 TI - [Parathyroid hormone-related protein. A new protein in search of a function]. PMID- 8728780 TI - [The real significance of basic research and the present status of CONICET]. PMID- 8728781 TI - [Chronic obstructive pulmonary disease. The Argentine Consensus]. PMID- 8728782 TI - [The Brazilian Digestive Motility Society and Arquivos de Gastroenterologia]. PMID- 8728783 TI - [Ribavirin for chronic hepatitis C: many doubts and some hope]. PMID- 8728784 TI - [Ribavirin in the treatment of hepatic cirrhosis due to hepatitis C virus. A non controlled study]. AB - Ribavirin in a fixed doses of 1.0 g/day was administered to 15 cirrhotic patients with portal hypertension and HCV-RNA detected in the blood by PCR during a six months period, in a non-controlled study. Two patients presented complications due to hepatic insufficiency and their data were not available at the end of the study. In four patients the hemoglobin levels fell below 20% of the initial value and in one the ribavirin dose had to be reduced. No other significant adverse reaction to the drug was observed. Almost all patients experienced a decrease in the aminotransferases levels during the study, specially in the first three months of treatment when the AST and ALT levels were significantly reduced when compared with the initial values. At the end of six months, four patients presented a complete response with normal aminotransferases levels, but only in one patient the HCV-RNA was not detected in the blood. In this patient the drug interruption was followed by elevation in aminotransferases levels and HCV-RNA detection in the blood 45 days later. Such results suggest that although well tolerated and inducing a transient decrease in AST and ALT, ribavirin alone administration is not able to erradicate virus C infection in patients with hepatic cirrhosis and portal hypertension. PMID- 8728785 TI - [Endoscopic therapy of pancreatic pseudocyst]. AB - Endoscopy therapy of pancreatic pseudocyst has been described as an efficient method. We report six cases of pancreatic pseudocyst treated by pancreatic stenting (three cases), cystogastrostomy (three cases) and cystoduodenostomy (one case). Rapid symptomatic improvement and pseudocyst regression were noted in all cases, except one. Such patient needed pancreatic stenting even after cystogastrostomy due to main pancreatic duct stenosis. There were three complications: hemorrhage (one case) and pseudocyst infection (two cases), and all were treated clinically. Two patients had pseudocyst infection previous to endoscopic manipulation, and they were treated only with antibiotics and endoscopic drainage, without surgery (treatment suggested by most of the authors). We concluded that endoscopic therapy of pancreatic pseudocyst is efficient and safe, being responsible for rapid relief of symptoms in most of patients. PMID- 8728786 TI - [Hepatic artery aneurysm: treatment with left hepatectomy. Report of a case]. AB - A 71-year-old female with a rupture of left intrahepatic artery aneurysm with acute upper abdominal pain, weight loss and fever. The diagnosis was established with doppler ultrasound, contrasted abdominal computer tomography scanning and celiac and mesenteric artery angiography. Prompt recognition and left hepatectomy led to a favourable outcome. PMID- 8728787 TI - [Inflammatory bowel disease activity index: clinical and laboratory indicators]. AB - The quantification of the degree of activity of inflammatory bowel disease is assuming growing importance nowadays. The activity index of the disease can be attained by clinical and laboratorial indicators. For ulcerative colitis the mostly used clinical parameters are daily bowel movements and presence of bloody diarrhea whereas albumin, hemoglobin, ESR and positive acute phase protein measurements are the laboratory parameters. For Crohn's disease activity besides the daily bowel movements the presence of abdominal pain and discomfort sensation are also frequently used whereas the C-reactive protein is the most used laboratory test which is able to detect the disease reactivation even before the appearance of any clinical sign. The combinations of clinical signs with the laboratory tests earned the sympathy of the specialists and the set of ensembled indicators has been recognized by the author's name. In this sense, the classification of the ulcerative colitis activity originally proposed by Truelove and Witts deserves presently a wide acceptance whereas such agreement is still lacking for Crohn's disease activity. In the mean time, the Bristol index is clinically the most feasible, once the Crohn's disease activity index and the Van Hees index are considered too complex. However the latter indexes are still useful mainly for comparisons among multicentric data. It seems that the currently existing clinical signs used for Crohn's disease activity would be quantitatively improved by adding some easily made laboratory tests such as C reactive protein. PMID- 8728788 TI - [Ethanol and the gastrointestinal tract]. AB - The effects of ethanol upon the gastrointestinal tract (mouth, pharynx, esophagus, stomach, duodenum, Oddi's sphincter, small bowel, colon and rectum) were reviewed. Several studies showed that the incidence of cancer in the mouth and pharynx is increased in alcoholics as a consequence of ethanol effects and probably those of other compounds found in liquors. The gastroesophageal reflux disease may be induced by alcohol since it reduces the pressure in the lower and the upper esophageal sphincter, as well as the extent of primary peristalsis. Several studies showed a strong correlation between esophageal cancer and alcohol abuse. The risk for developing this kind of tumour is significantly increased when alcohol abuse and smoking coexist. Alcoholism predisposes patients to Mallory-Weiss syndrome as well as to bleeding of esophageal varices Ethanol may affect gastric secretion, motility, and permeability. Some drugs acting upon the gastric alcohol-dehydrogenase are able to affect gastric absorption of ethanol. Eradication of Helicobacter pylori increases the activity of alcohol dehydrogenase in the pyloric antrum. The effects of alcohol upon the gastric mucosa include caustic damage, retrograde diffusion of H+, and cytoprotection. This agent may cause an acute gastritis but it is probably not involved in chronic gastritis. Whether alcohol is a risk factor for ulcer or not is unknown. Some studies found an increased incidence of gastric cancer associated with consumption of beer, wine and vodka. Some authors reported a decreased pressure in Oddi's sphincter while others found it increased in association with the consumption of ethanol. The acute and the chronic consumption of alcohol may affect the structure of small bowel as well as the absorption of nutrients. Several studies reported a significant correlation between colorectal cancer and the chronic consumption of ethanol. PMID- 8728789 TI - [Steatocrit and Sudan III in the study of steatorrhea in children: comparison with the Van de Kamer method]. AB - Steatorrhea was investigated in 50 children, through Van de Kamer test, Sudan III and steatocrit, in order to compare these three methods. Steatocrit showed better correlation with Van de Kamer test, obtaining sensibility and specificity of 91% and 87%, respectible, on stools took before homogenizing the feces collected during 72 hours. In this same situation, Sudan III showed sensibility of 73% and specificity of 69%. As in the Van de Kamer test, steatocrit represents the final expression of all fecal lipids, what does not happen with Sudan III. PMID- 8728791 TI - Ultrastructural study of enteropathogenic Escherichia coli O111ab:H2 infection in an infant with acute diarrhea. AB - Enteropathogenic Escherichia coli is the most important cause of acute diarrhea in developing countries, specially in infants under one year of age. Enteropathogenic Escherichia coli strains are able to induce profound cytoskeletal alterations in the enterocyte known as attaching and effacing lesions, associated with the formation of cuplike pedestals. We report an Enteropathogenic Escherichia coli O11ab:H2 strain isolated from an infant with acute diarrhea, on the eleventh day of disease, that caused attaching and effacing lesion and penetrated the enterocyte, as well as invaded the HeLa cell tissue culture in vitro and the rabbit ileal loop assay in vivo, in the ultrastructural study. This observation indicates that the severe lesions of the small bowel caused by an enteropathogenic Escherichia coli O111ab:H2 strain can occur even in the early stages of the infection. PMID- 8728792 TI - On the statistical distribution of amino acid residue in randomly synthesized peptides. AB - The probability of randomly synthesized peptides having an excess of a given residue Ri (nRi > N/2; N = size of the peptide) decreases strongly with peptide size. For a strong specific interaction of a Ri, rich peptide with a given sequence of a ribotide chain, peptides should be reasonably large. We discuss how a compromise can be achieved that may have had an important role on the origin of the genetic code. PMID- 8728790 TI - [Hepatic glycogenosis in childhood: clinical and laboratory findings in 20 patients]. AB - We studied 20 children with a clinical picture and laboratory study suggestive of hepatic glycogenosis. The age of the beginning of symptoms varied from birth to 24 months and the age at the diagnosis varied from 2 to 81 months. Hepatomegaly was found in all patients, diarrhea in 65% (13/26), "doll-face" in 55% (11/20) and convulsions in 50% (10/20). Nutritional evaluation showed more height deficiency than weight deficiency. Laboratory tests showed elevation of hepatic transaminases (12/19), hypercolesterolemia (8/14), hyperuricemia (6/17) and hypoglycemia (6/20). Liver function was not compromised in most of the cases. The results of glucagon tolerance test were variable. The histoenzymology study performed in 15 patients revealed the following results: Type VI (liver phosphorylase deficiency) in seven, Type I (glucose-6-phosphatase deficiency) in two, Type IV (brancher enzyme) in one and no conclusion could be drawn in five patients. The finding of hypoglycemia in few cases of this study can be justified by the few number of glycogenosis Type I, probably due to the fact that this type is the most easily diagnosed, with less necessity of referring them to specialized centers. PMID- 8728793 TI - Ionization potentials and electron affinities of nicotinic acid and nicotinamide calculated with HAM/3. AB - Ionization potentials and electron affinities of nicotinic acid and nicotinamide were calculated by HAM/3. Observed photoelectron spectra of the molecules were analyzed with the aid of the calculated ionization potentials. Chemical reactivity of the molecules was discussed. PMID- 8728794 TI - [Ethos, psyche, soma. Medicine and its mutations]. AB - In this paper, understood as a contribution to a theoretical pathology, the necessity and urgency of a medical metatheory are explained on the basis of intensive and extensive developments of medical knowledge. These developments have extended beyond the limits of medicine modelled after the experimental and clinical paradigms, making it imperative to include the social and valoric dimensions within medicine and not as an addition to its traditional goals. The metamedicine so created, it is suggested, should develop a dialogic model in which, without having to postulate isomorphisms or correlations, the dilemmas posed by ethos, psyche and soma are confronted as discourses of a human science. PMID- 8728795 TI - [Psychic self-activity: history and currently of a construct]. AB - The psychic self-activity is a hypothetical construct meant to describe and explain the mental processes which are not entirely generated by environmental stimuli nor by the need of eliminating internal stimuli. It emphasizes the proactive behavior led by curiousity, the exploratory urge and the psychic drive increase. Initially conceived by philosophers and the intuitive psychologists, this construct tends to disentangle from that speculative past and to focus on the empirical research. PMID- 8728796 TI - [Application of the Michigan Alcoholism Screening Test in heads of the family]. AB - The habit of alcohol ingestion was examined in major caretakers of extreme poverty families who live in a county of the southern area of Santiago, Chile. For this purpose we used the Michigan Alcoholism Screening Test (MAST) validated in Chile. This test allows the disrimination between normal and abnormal alcohol drinkers. In this study the prevalence for abnormal drinkers was 40.3% with a 46.2% of prevalence if the major caretaker was the male parent and a 3.3% of prevalence if the major caretaker was a female one. This prevalence was studied associated with the following variables: socioeconomic stratification index (CAS), education, occupation, number of family members and neighborhood district. PMID- 8728797 TI - [Schizophrenia: relationship between negative symptoms and schizotypic features of personality]. AB - We studied the negative and schizoid-schizotypic features in 19 males chronics schizophrenic inpatients. Our findings prove a direct relation between negative symptoms and schizotypical personality features. We think that the negative symptoms not form a unity and one of them are part of the prior personality. PMID- 8728798 TI - [The residents of mental health services in 1994. Population characteristics and its opinions about graduation]. AB - Departing from two surveys that took place during a Mental Health Meeting of the Health Residents from Buenos Aires, the main features and their opinions upon their education and work are analyzed. The results are discriminated by profession. Two problems are highlighted: one of them refers to the general insatisfaction of this group with their theoretical learning; the other, shows the exit of prepared human resources from public hospitals. The data suggest that, despite sharing most of their activities, psychologists and psychiatrists have significant different opinions about the clinic education they think are getting. Further research is suggested in order to have a deeper comprehension of what is going on in these training programs. PMID- 8728799 TI - [Platelet serotonin in endogenous depressive patients and normal controls]. AB - We have studied the content of platelet serotonin in 31 depressive patients free of medication and in 29 controls. The depressive patients have been selected according to clinic diagnosis and to urinary phenylethylamine (PEA) and total phenylacetic acid (PAA) values, because low levels of these two substances are for us almost a constant in endogenous depressive patients. For the control group we selected 23 out of 29 cases studied, which had all urinary PEA and PAA values within the normal range published by us previously. In this group platelet serotonin content was of 527-900 ng/10 9 platelets with a mean of 698 +/- 125 ng/10 9 platelets. In the depressive group, taking account of platelets serotonin content we could differentiate two subgroups. In a group of 19 patients we found lower values, 157-513 ng/10 9 platelets with a mean of 407 +/- 102 ng/10 9. In the other 13 patients group we found platelet serotonin levels similar to those of the control group, 552-780 ng 10 9 platelets with a mean of 658 +/- 93 ng/10 9 platelets. Accepting blood platelets as models for serotoninergic nerve terminals, we suppose that the response of depressive patients to serotonin reuptake blockers treatment should be different in both subgroups, which is at present matter of study in our laboratory. PMID- 8728800 TI - [The beginning of psychology as a profession in Argentinian universities]. AB - Two characteristics of professional psychology in Argentine are interesting for historians. On the one hand, the omission of the large past of scientific psychology that began at the end of last century. On the other, the omission of the most recently beginning of professional psychology on '50. In this paper, we point out some political features of the process of psychology's professionalization around those latter years. The main subject of this paper, are the studies of psychology at Tucuman and Cuyo universities, on '40 and '50. In those years, the social and industrial changes in the country, leads to new problems of adaptation to job and to environment. Those subjects and the old problems of psychological aspects in school, pave the way for the new needs of psychological profession in all the country. PMID- 8728801 TI - [Mediastinal paraganglia lesions in chronic chagasic heart disease: cause of sudden death?]. PMID- 8728802 TI - [Strategies of early reperfusion in acute myocardial infarction]. AB - PURPOSE: To describe the use of myocardial reperfusion strategies (percutaneous transluminal coronary angioplasty-PTCA and intravenous trombolysis) whenever it is possible to use emergency cinecoronariography in acute myocardial infarction (AMI). METHODS: The records of patients admitted with initial diagnosis of AMI, until six hours after the beginning of symptoms, were reviewed retrospectively, between March/92 and December/93. RESULTS: One hundred forty-three patients were admitted with suspected AMI. Eighty-one (57%) presented definitive criteria for this diagnosis. Fifty-two patients (64%) were admitted in the first six hours from the beginning of symptoms. Thirty-eight (73%) were male and the mean age was 59 +/- 12 years. Cinecoronariography was indicated immediately in 36 of 52 (69%) patients, with purpose to use PTCA. That was done in 30 (58%). Two (4%) patients were submitted to emergency surgical myocardial revascularization. Intravenous thrombolysis was used in 11 (21%) patients. At admission, the conservative treatment was chosen for five out of nine left patients, based on clinical grounds. CONCLUSION: The availability of emergency cinecoronariography made an early reperfusion strategy possible in 83% of patients admitted with AMI in the first six hours after the beginning of symptoms. PMID- 8728803 TI - [Intracoronary isosorbide mononitrate versus placebo. A double-blind study with quantitative coronary angiography]. AB - PURPOSE: To evaluate the vasodilation effect of intracoronary isosorbide mononitrate (ISMN) on the coronary arteries, systolic aortic pressure (SAP) and cardiac rhythms. METHODS: Forty patients aged 61 (40-72) years, 30 (75%) male, were randomized to receive on a double blind fashion, 20 mg of ISMN or placebo. The two groups had similar baseline characteristics. Quantitative coronary arteriography [by cardiovascular measurement system (CMS)] and SAP were compared before and after ISMN. We also observed the cardiac rhythm. RESULTS: There were no arrhythmias. Reference vessel diameter increased from 2.6 +/- 0.5 mm before to 2.9 +/- 0.4 mm after ISMN and did not change, mean 2.9 +/- 0.5 mm, before and after placebo. There was a difference between ISMN and placebo (p < 0.0001). There was no difference in the SAP change between the two groups, 4.2 +/- 0.7 mmHg with ISMN and 1.8 +/- 0.5 mmHg with placebo (p = NS). CONCLUSION: ISMN promotes a safe and effective vasodilation of the coronary arteries with no major effects in cardiac rhythm and systolic aortic pressure. PMID- 8728804 TI - [Is psychoprophylaxis a valid approach for heart surgery in children?]. AB - PURPOSE: To validate the hypothesys that a psycological preparation for children who will undergo cardiac surgery may improve the outcome. METHODS: Sixty patients, with ages ranging between 3 and 10 years, submitted to heart surgery for treatment of congenital heart defects, were evaluated. They were divided in 2 groups: experimental and control. A questionnaire was designed for collecting data about psychological and clinical aspects of each patient. RESULTS: The following data was found to be of statistical significance: acceptance of peripheral vein puncture in the surgical group (chi 2 = 11.59, p < 0.05), calm awakening following general anesthesia (chi 2 = 9.64 p < 0.05), cooperation with the physiotherapy staff (chi 2 = 13.30, p < 0.05), coping with parents absence (chi 2 = 9.64, p < 0.05), acceptance of fluid restriction (chi 2 = 17.78, p < 0.05) and cooperation with removal of stitches and pacemaker electrodes (chi 2 = 19.20, p < 0.05). There was not statistical significance on demand of sedation, cooperation at removal of the orotracheal tube and during examination, necessity of reintubation and occurrence of clinical complications. However, the prepared group showed a slight tendency to have less postoperative complications (20%) than the control (27%). CONCLUSION: It was found that children who had adequated psychologic preparation prior to the correction of congenital heart defects had better psychological results with the imposed trauma. PMID- 8728805 TI - [Epidemiology of rheumatic fever. A local study]. AB - PURPOSE: To identify the clinical findings of first attack in acute rheumatic fever (ARF), and to find a profile of its epidemiology. METHODS: Results of clinical manifestations, incidence, laboratory findings relative to a new cases of first attack in ARF were studied. Six hospitals in Porto Alegre area were involved in the surveillance, during 16 months between 1992 and 1993. RESULTS: Fifty-one cases with clinical manifestations were recorded: 40 (78.4%) cases with arthritis; 28 (56%) with carditis; 6 (11.8%) with Sydenhan's Chorea, 6 (11.8%) with erythema marginatum and 4 (7.8%) with subcutaneous nodes. CONCLUSION: Diagnostic criteria firstly described by Jones are still employed in the clinical diagnosis in ARF in a first attack and that remains the principal etiology of valvar heart disease in that city. PMID- 8728806 TI - [Anomalous venous drainage of the left lung. Clinical and surgical findings and diagnostic difficulties]. AB - PURPOSE: To analyze clinical and surgical aspects of patients with anomalous left pulmonary venous drainage (ALPVD). METHODS: Seven patients, 3 males, with ages ranging from 18 months to 29 years were retrospectively studied, by analyzing the symptoms, electrocardiograms, chest X-ray, echocardiograms, angiography and surgical technique. RESULTS: All patients, but one, were symptomatic being the effort dyspnea the most prevalent symptom. Electrocardiogram showed incomplete right bundle branch block in all but one case. Chest X-ray showed prominent pulmonic vessels (6 patients), different degrees of cardiomegaly (6) and a left superior vena cava like image (3). Echocardiography correctly depicted the anomaly in 4 cases. The ALPVD was lobar in 4 cases and total in 3. In 3 patients there was pulmonary hypertension and in one venous drainage obstruction (8mmHg gradient). Six patients were operated on, 3 through a sternotomy approach with cardiopulmonary bypass and other 3 through a left thoracotomy without cardiopulmonary bypass. CONCLUSION: ALPVD has varying clinical repercussion and its clinical picture resembles atrial septal defect. Some cases may present diagnostic difficulties and echocardiograms may result in false negative findings. Cardiac catheterization is important to obtain detailed anatomic diagnosis, to evaluate the degree of pulmonary hypertension and to rule out venous obstruction. Left thoracotomy without extracorporeal circulation is indicated in ALPVD without associated defects. PMID- 8728807 TI - [Prevalence of rheumatic fever in children from a public high school in Belo Horizonte]. AB - PURPOSE: To determine the prevalence of rheumatic fever (RF) among children of a public high school in Belo Horizonte, Brazil. METHODS: The study was performed from March to December/92, and involved high school students coming from families of the medium and low-medium social classes. Considering the total of 1,400 students registered in a public school and the estimated RF prevalence in the developing country, 729 students were randomly chosen to be interviewed and examined by a researcher. The children suspected of being affect by RF were submitted to echocardiography in order to find any cardiac lesions. RESULTS: Due to several different factors, only 550 students aging from 10 to 20 years were admitted in this study. Four children, previously under secondary prophylactics, had already had the diagnosis of RF. Among these children, only one had heart disease (combined mitral incompetence and stenosis, and aortic incompetence). From the other three children, only one had the diagnosis of RF confirmed according to the Jones criteria. CONCLUSION: From the ata obtained, the prevalence of RF was calculated in 3.6/1000. PMID- 8728808 TI - [Anomalous origin of the left coronary artery. Concomitant correction with valvar replacement for mitral insufficiency]. AB - A four year-old boy with an anomalous origin of the left coronary artery (LCA) directly from the right pulmonary artery was reported. He had been underwent to mitral valve replacement by a St Judes no. 29, due to severe mitral insufficiency, secondary to an extensive myocardial infarction including the anterior and posterior papilary muscles, beside the implantation of the LCA in the aorta. Instead of this favorable outcome, it's emphasized the need for earlier recognition to avoid risk sequelae. PMID- 8728809 TI - [Case 5/95 (Hospital Universitario Antonio Pedro - Universidade Federal Fluminense)]. PMID- 8728810 TI - [Cardiopulmonary involvement in schistosomiasis]. PMID- 8728811 TI - [Thrombolysis versus angioplasty in acute myocardial infarction]. PMID- 8728812 TI - [Strategies of early reperfusion in acute myocardial infarction]. PMID- 8728813 TI - [Natural history of Chagas disease]. PMID- 8728814 TI - [Pathogeny of cardiac manifestations in Chagas disease]. PMID- 8728815 TI - [Ventricular arrhythmia in Chagas disease. Diagnostic, prognostic, and therapeutic features]. PMID- 8728816 TI - [Chagas disease in the context of organ transplantation]. PMID- 8728817 TI - Brain, language and the origin of human mental functions. AB - We propose that to understand the biological and neurophysiological processes that give rise to human mental phenomena it is necessary to consider them as behavioral relational phenomena. In particular, we propose that: a) these phenomena take place in the relational manner of living that human language constitutes, and b) that they arise as recursive operations in such behavioral domain. Accordingly, we maintain that these phenomena do not take place in the brain, nor are they the result of a unique operation of the human brain, but arise with the participation of the brain as it generates the behavioral relational dynamics that constitutes language. PMID- 8728818 TI - Sexual dimorphism in interhemispheric relations: anatomical-behavioral convergence. AB - An embryogenetic hypothesis states that hemispheric specialization is inversely related to callosal connectivity (Geschwind and Galaburda, 1985). We tested this hypothesis (i) anatomically by relating postmortem planum temporale asymmetry to regional callosal morphology and (ii) behaviorally by relating the right visual field advantage in a lateralized lexical decision task with associative primes to regional callosal morphometry using magnetic resonance imaging (MRI). The postmortem study showed a significant negative correlation between planum temporale asymmetry and the number of small diameter fibers in the isthmus of the corpus callosum, but only for males. The MRI study showed a significant negative correlation between the right visual hemifield advantage for associated words and the cross section size of the isthmus of the corpus callosum, but again only in males. There was no sex difference in either the anatomical asymmetry, the behavioral asymmetry, or the callosal morphology. These convergent results suggest that there is a sexual dimorphism in interhemispheric relations in humans. PMID- 8728819 TI - The anatomical substrates for language and hemispheric specialization. AB - Three main lines of investigation are discussed in this paper: (1) the comparison between the anatomical arrangement of the language areas and the large-scale neurocognitive cortical networks partly involved in active or working memory; (2) the relations between hemispheric specialization and the development of interhemispheric communication; and (3) the analysis of individual differences in brain organization for language. The hypothesis and evidence presented stem from work being performed in our laboratories. PMID- 8728820 TI - The case for a relationship between human memory, hippocampus and corpus callosum. AB - Unilateral brain damage which includes the hippocampus leads to memory impairments consistent with hemispheric specialization on the same side. Damage to the corpus callosum, the major connecting pathway between the left and right hemispheres, also leads to memory impairments. This suggests both hemispheric specialization on the hippocampal level and a critical role for the corpus callosum in memory functions. A complete hippocampal formation is present on either side of the brain but traditionally only one is studied. However, a comparison between the neuronal populations in the hippocampus on both sides revealed asymmetry in connectivity among hippocampal subfields. The profile of memory impairments of commissurotomy ('split-brain') patients is described. The results are discussed in terms of a relationship between hippocampus and corpus callosum in humans. As hemispheric specialization evolved, inter-hippocampal connections became less important and the corpus callosum became prominent in memory functions. PMID- 8728821 TI - Memory in the cortex of the primate. AB - Memory is viewed as hierarchical and distributed in primary and association areas of cerebral cortex. Different memory neural networks are interconnected at various levels in this hierarchy, sharing neurons and connections. All memory is essentially associative in its generation, structure and retrieval. External and internal stimuli, to which we attend by virtue of their biological relevance or for other reason, can at any time activate ("turn on") the neuronal network to which they belong by previous association. This is the basis of knowledge and remembering. The reverberation in recurrent circuits may keep the network in an active state, that is, serving behavior, attention and consciousness. Monkey neuropsychological and electrophysiological data, and human tomographic (brain metabolism) evidence are presented supporting these concepts. PMID- 8728822 TI - Elementary forms of synaptic plasticity in the visual cortex. AB - The neocortex is an important site of memory storage, and memories are believed to be formed in the cortex by the activity-dependent modification of synaptic connections. However, in contrast to the hippocampus where there has been an increasingly sophisticated analysis of synaptic plasticity, relatively little is known about the mechanisms of synaptic modification in neocortex. Here we summarize the results of a series of experiments conducted on slices of visual cortex in vitro, aimed at elucidating the elementary mechanisms of synaptic plasticity in the superficial layers of neocortex. We show that long-term potentiation (LTP) and depression (LTD) result from high- and low-frequency conditioning stimulation, respectively, of the middle layers of cortex. Both forms of synaptic plasticity are input-specific and dependent on activation of postsynaptic N-methyl-D-aspartate (NMDA) receptors. The critical variable in determining the sign of the synaptic modification appears to be the level of postsynaptic depolarization during conditioning stimulation. The data support a model in which the state of correlation of pre- and post-synaptic activity is converted by the voltage-dependent NMDA receptor channel into a graded postsynaptic Ca2+ signal. LTD is triggered by a modest but sustained elevation in postsynaptic Ca2+, while LTP is elicited by larger changes in Ca2+. An important variable that regulates synaptic plasticity in the neocortex is intracortical inhibition, which constrains the patterns of activity that can reach the modifiable synapses in layer III. PMID- 8728824 TI - Linear analysis of auto-organization in Hebbian neural networks. AB - The self-organization of neurotopies where neural connections follow Hebbian dynamics is framed in terms of linear operator theory. A general and exact equation describing the time evolution of the overall synaptic strength connecting two neural laminae is derived. This linear matricial equation, which is similar to the equations used to describe oscillating systems in physics, is modified by the introduction of non-linear terms, in order to capture self organizing (or auto-organizing) processes. The behavior of a simple and small system, that contains a non-linearity that mimics a metabolic constraint, is analyzed by computer simulations. The emergence of a simple "order" (or degree of organization) in this low-dimensionality model system is discussed. PMID- 8728823 TI - Resonant cell assemblies: a new approach to cognitive functions and neuronal synchrony. AB - This paper presents a novel reading of ideas on temporal binding as a key for cognitive operations by means of fast (gamma band) phase synchrony. We advocate a view of binding of widely distributed cell assemblies transiently locked in a neural hypergraph which serves as a reference point to incorporate or interpret other less coherent concurrent neural events. The paper traces in some detail the empirical evidence concerning the gamma binding process and presents some implications for the constitution of a unified cognitive-mental space. PMID- 8728825 TI - Central noradrenergic hyperactivity early in life: a hypothesis on the origin of morpho-functional brain disorders induced by malnutrition. AB - Many studies have revealed that malnutrition, caused either by insufficient or unbalanced diet, during early stages of growth and development could result in a variety of morpho-functional brain disturbances, whose severity depends on the time of onset, duration and intensity of the nutritional injury. Nevertheless, little is known about the intimate mechanisms by which early malnutrition impairs brain structure and function. This article reviews evidence showing that (i) developmental malnutrition induces central noradrenergic hyperactivity, (ii) noradrenaline exerts a trophic role during brain development, and (iii) pharmacological reduction of central noradrenergic hyperactivity prevents malnutrition-induced functional brain disturbances. PMID- 8728826 TI - The activation of bulbo-spinal controls by peripheral nociceptive inputs: diffuse noxious inhibitory controls. AB - Some neurones in the dorsal horn of the spinal cord are strongly inhibited when a nociceptive stimulus is applied to any part of the body, distinct from their excitatory receptive fields. This phenomenon was termed "Diffuse Noxious Inhibitory Controls" (DNIC). DNIC influence only convergent neurones, and these inhibitions can be triggered only by conditioning stimuli which are nociceptive. The inhibitions are extremely potent, affect all the activities of the convergent neurones and persist after the removal of the conditioning stimulus. Only activity of A delta- or A delta- and C- peripheral fibres can trigger DNIC. DNIC are sustained by a complex loop which involves supraspinal structures since, unlike segmental inhibitions, they are not observed in animals in which the cord has previously been transected at the cervical level. The ascending and descending limbs of this loop travel respectively through the ventro-lateral and dorso-lateral funiculi, respectively. We proposed that DNIC result from the physiological activation of some brain structures putatively involved in descending inhibition. However, lesions of the mesencephalon, including the periaqueductal grey (PAG) and the rostral ventromedial medulla (RVM), including nucleus raphe magnus, did not modify DNIC. By contrast, lesions of subnucleus reticularis dorsalis (SRD) in the caudal medulla strongly reduced DNIC. Both electrophysiological and anatomical data support the involvement of SRD neurones in spino-bulbo-spinal loop(s). In man, very similar results have been obtained by means of combined psychophysical measurements and recordings of nociceptive reflexes (RIII reflex). Painful heterotopic conditioning stimuli depress both the reflex and the associated painful sensation, with stronger effects being observed with more intense conditioning stimuli. By contrast, in tetraplegic patients, heterotopic nociceptive stimulation did not produce any depression of the RIII reflex. Observations were also made on patients with cerebral lesions causing contralateral hemi-analgesia, either a unilateral thalamic lesion or a lesion of the retro-olivary part of the medulla (Wallenberg's syndrome). In the patients with Wallenberg's syndrome, no inhibitions were observed when the nociceptive conditioning stimuli were applied to the affected side whereas if these stimuli were applied to the normal side they triggered inhibitory effects and post effects very similar to those seen in normal subjects. These results show that in humans, brainstem--probably reticular--structures seem to play a key role in these phenomena. The data suggest that nociceptive stimuli, even though there are unquestionably perceived as being painful activate certain inhibitory controls which originate in the brainstem. Since all convergent neurones are subject to DNIC, one can make the assertion that the transmission of nociceptive signals towards higher centres is under the influence of these controls. In other words, the descending inhibitory controls may play a physiological role in the detection of nociceptive signals. It is proposed that DNIC constitute both a filter which allows the extraction of the signal for pain and an amplifier in the transmission system which increases the potential alarm function of the nociceptive signals. This hypothesis is supported by the finding that DNIC are blocked by low doses of morphine in both rat and man. PMID- 8728827 TI - A report on the evolution of this journal and an analysis of its present state and perspectives. AB - This journal appeared as Archivos de Biologia y Medicina Experimentales between 1964 and 1992, its name being changed into Biological Research in 1992. This is a report on the past and present of the journal, analyzing the problems of a scientific journal published in English but produced in Latin America, those that its contributors must deal with in the preparation of the manuscripts, those involved in the evaluation procedures and editing of material for the press, and those about coverage by indexing services and the international recognition of the journal quality. PMID- 8728828 TI - Determination of the pKa of ionizable enzyme groups by nonlinear regression using a second degree equation. AB - A second degree equation fitted by nonlinear regression for the analysis of the pH effect on enzyme activity is proposed for diprotic enzyme systems. This method allows the calculation of two molecular dissociation constants (KE1 and KE2 for the free enzyme, KES1 and KES2 for the ES complex) and the pH independent parameters (Vmax and Vmax/Km). The method is validated by bibliographic (alpha chymotrypsin) and experimental data (almond beta-D-glucosidase). No significant differences were found between present data and those previously reported in the literature using similar experimental conditions. This method works using comparatively few [H+] concentration values within a narrow pH range, preferentially around the optimum, being adequate for diprotic systems with close pKa values. PMID- 8728830 TI - Preparation, production and functional characterization of anti cAMP and cGMP antibodies. AB - The present work was intended to evaluate the preparation of antigens, as well as the production and characterization of anti cAMP and cGMP antibodies. Such antibodies were obtained from rabbits, and we used 2'O-succinyl cyclic nucleotide derivate, conjugated with human serum albumin, as antigen. The characterization of the antibodies was monitored by their immunoreactivity with the labelled antigen [125I]-cyclic nucleotide. This assay consists in a competition between a labelled and an unlabelled antigen for a fixed number of binding sites present in the specific antibody. The antibodies were specific for the inducing antigens. Cross-reactivity tests showed low degree competition between the immunogen and other antigens. The very high affinity, high quality and specificity of the generated antibodies indicate that they may be used not only in radioimmunoassay and immunocytochemistry methodologies, but also as bioblockers of physiological pathways. PMID- 8728829 TI - Differentiation of oxyntic cells and cell-matrix interactions during avian gastric gland morphogenesis. AB - The relation between the expression of the oxyntic cell phenotype and the modifications of the extracellular matrix during development of the gastric glands, was studied in 10 to 21 day-old chick embryos. Cytodifferentiation of the oxyntic cells was established by ultrastructural methods, while the expression of pepsinogen, mitochondrial enzyme markers and apical secretory membranes was determined by histochemical and biochemical procedures. Results show that the morphogenesis of the glandular lobules occurs between days 8 and 15 of gestation. Later on, the lobules enlarge but maintain their basic morphology. Until day 13, the developing glands consist of primary tubes lined by a stratified columnar epithelium. The apical poles of the cells that contact the lumen show cytoplasmic processes, and Mg-ATPase activity and F-actin are concentrated at the apical cell borders. From day 13 on, the cells of the simple epithelium that lines secondary tubules budding from the primary tube, show all the features that define differentiated oxyntic cells. The synthesis of glycosaminoglycans during glandular morphogenesis was studied measuring the incorporation of radioactive sulfate into developing chick embryo proventriculi. An important increase in isotope incorporation was found between days 13 and 18 of development. Histochemical localization of these macromolecules shows that glycosaminoglycans are closely associated with the developing glandular lobules. Variations in the structure of epithelial cells undergoing morphogenesis and in the composition of the extracellular matrix are synchronous, suggesting that interactions between them may be significant in terms of the establishment and maintenance of the adult gastric gland phenotype. PMID- 8728831 TI - Policosanol inhibits cholesterol biosynthesis and enhances low density lipoprotein processing in cultured human fibroblasts. AB - Policosanol is a mixture of aliphatic primary alcohols isolated and purified from sugar cane wax, that induces cholesterol-lowering effects in experimental models and human beings. When human lung fibroblasts were incubated with policosanol for 48 hours prior to the experiment, a dose dependent inhibition of 14C-acetate incorporation into total cholesterol was observed, whereas labeled mevalonate incorporation was not inhibited. Even when cholesterol synthesis was not strongly inhibited, low density lipoprotein (LDL) processing was markedly enhanced. Thus, LDL binding, internalization and degradation were significantly increased after policosanol treatment. In addition, despite the fact that'cholesterol generation was not inhibited at the lowest dose of policosanol assayed, LDL processing was significantly increased. The current data indicate that policosanol inhibits cholesterol synthesis at the earliest steps of the cholesterol biosynthetic pathway. On the other hand, this study suggests that the increase in LDL processing may be partially explained by the inhibition of cholesterol biosynthesis, even though an sterol-independent mechanism might be responsible for the enhancement of LDL-receptor activity. PMID- 8728832 TI - Cholesterol-lowering effects of policosanol in rabbits. AB - Policosanol is a natural mixture of higher primary aliphatic alcohols isolated and purified from sugar cane (Saccharum officinarum, L.) wax, whose main component is octacosanol. Policosanol (5-200 mg/kg) orally administered for 4 weeks to normocholesterolemic New Zealand rabbits significantly reduced total cholesterol and low density lipoprotein cholesterol (LDL-C) serum levels in a dose dependent manner. Serum triglyceride levels of treated and control animals were significantly different, but the reduction observed was not dose-dependent. High density lipoprotein cholesterol (HDL-C) levels remained unchanged. Results indicate that the reduction in total cholesterol values induced by policosanol is mainly mediated through a decrease in LDL-C levels. PMID- 8728833 TI - Influence of the estrous cycle on the norepinephrine-induced contraction of rat aorta: relationship to vascular prostanoids biosynthesis. AB - Since ovarian sex steroids (estradiol and progesterone) may affect both blood pressure and prostanoids synthesis, and because prostaglandin-E2 (PGE2) and prostacyclin (PGI2) can modulate the vascular action of pressor hormones, we investigated the vascular reactivity to norepinephrine during the estrous cycle of the rat. In addition, we determined the vascular biosynthesis of PGE2 and 6 keto-PGF1 alpha (the stable metabolite of PGI2) at different stages of the estrous cycle. Cumulative dose-response curves were obtained by a stepwise increase in the concentration of norepinephrine. The contraction of thoracic aortic rings induced by norepinephrine did not change significantly between estrus, metestrus and diestrus. However, aortic rings obtained on proestrus showed a significant reduction in the maximal contraction (Emax) induced by norepinephrine (p < 0.001). In addition, we found significant increases in vascular synthesis of PGE2 and PGI2 on proestrus (p < 0.001). These results indicate that vascular reactivity and vascular prostanoids synthesis are influenced by the hormonal changes occurring during the estrous cycle of normal female rats. It is possible that prostanoids generated locally may play an important role in the regulation of vasomotor tone in the systemic vascular bed throughout the estrous cycle. PMID- 8728834 TI - Genotoxic effects induced in cultured Chinese hamster ovary (CHO) cells by contaminated aquatic environments. AB - The Bio-Bio river, running through one of the most important hydrographic basins in Chile, presents concentrations of some chemical agents exceeding the accepted values for continental aquatic environments. The area near to the mouth of the river is highly industrialized and the industrial effluents are discharge directly into the river, most of them without any previous treatment. This river provides the principal source of drinking water for a population of more than one million inhabitants in the region. To evaluate the genotoxic effects of liquid effluents from a cellulose industry and the surface waters of the Bio-Bio river obtained near to the river mouth in the proximity of Concepcion city, a short term bio-assay with cultured Chinese hamster ovary (CHO) cells was used. The frequency of cells with chromosome aberrations in metaphase, anaphase and telophase was determined at different concentrations of the liquid samples. The results show a significant increase in chromosomal damage. The frequency of chromosomal aberrations observed both in metaphase and ana-telophase is dose related to the concentrations of liquid samples tested. The superficial water shows a significant genotoxic effect. The scope of these results is discussed and compared to results obtained in other biological models. PMID- 8728835 TI - Physiological aspects of Trypanosoma cruzi gene regulation during heat-shock. AB - Investigations on the conditions of heat-shock response in Trypanosoma cruzi, the agent of Chagas disease, showed that at 37 degrees C, one of the heat-shock temperatures employed, the parasites from 48 h culture do not display a classical response to the heat treatment, since a general increase in RNA and protein synthesis was detected. The classical heat-shock response was detected only at 40 degrees C. The data also suggest that the heat shock proteins (HSP) mRNA population is sufficient to maintain protein synthesis at a high rate for at least 1 h and, to maintain the same rate of response for a longer period, transcription is necessary. The half life of HSP 70 mRNA is less than 3 h at 37 degrees C. The protein synthesized during the first hour of the heat shock at 37 degrees C is stable for at least 24 h. The parasite seems to be able to reuse the stock of HSP mRNAs stored during the first thermal shock to respond to a second heat treatment. These data are discussed bearing in mind other cell types. PMID- 8728836 TI - Mini-Mu technology in Salmonella typhi: isolation of stable MudJ operon fusions by cis complementation. AB - This paper describes a method to achieve stable MudJ insertions in the Salmonella typhi Ty2 chromosome. The method is a modification of the genetic complementation system described previously for Salmonella typhimurium, which consists in placing the defective transposon (MudJ) near the transposase genes of a helper Mu phage on a single DNA fragment. This fragment is then introduced into a new bacterial host by means of P22 transduction. We constructed a S. typhi strain which carries MudJ and the Mu helper phage in the chromosome. This strain was induced to lytic growth and the lysate was used to infect S. typhi Ty2. The frequency of mutation was 2.0 x 10(-6) mutants per recipient bacterium. Superinfection with the Mu helper phage was about 1%. To determine the number of MudJ insertions, several mutants were subjected to Southern blot analysis. From a total of 25 mutants analyzed, only 4 contained more than one insertion. Our procedure compares well with the method described previously for the S. typhimurium-P22 system and can be applied to other Mu sensitive bacteria. PMID- 8728837 TI - A new microassay for the determination of alkaline phosphatase activity in early mouse concepti. AB - A new enzymatic assay for the determination of alkaline phosphatase (APase) activity in preimplantation mouse concepti is described. This method allows estimation of APase activity of concepti extracts using the fluorogenic substrate, 3,6-fluorescein diphosphate (FDP). For measuring APase activity, 0.1% Nonidet P-40 was used to solubilize the enzyme. Control assays showed that this procedure does not modify the enzyme activity. According to the Km obtained for APase from mouse concepti (between 1-2 microM), the initial concentration of FDP was 20 microM, which is 10 fold the Km. The assay sensitivity allows continuous recording of the product generated and a reliable determination in less than 20 min. Results show that APase activity in mouse concepti may be detected from the 2-cell stage, increasing exponentially towards the blastocyst stage. PMID- 8728838 TI - Oxygen transfer in fish: morphological and molecular adjustments. AB - A wide range of organismic, physiological and biochemical adjustments to improve oxygen transfer is observed in fish exposed to environmental hypoxia and during anemia. Many fish species of the Amazon obtain oxygen directly from air when water oxygen is low. The accessory air-breathing organs include modifications of the gills, mouth, stomach and intestine, and swimbladder vascularization. Other species extend the lower lip and skim to improve oxygen uptake from the oxygen rich surface layer of the water. The amount of oxygen uptake from air was estimated for Hoplosternum littorale and Lipossarcus pardalis. In addition, the oxygen uptake from the water surface was estimated for Colossoma macropomum. Blood oxygen content was reduced by 30% in Hoplosternum littorale and Colossoma macropomum and 70% in Lipossarcus pardalis if they were denied access to air. Adjustments of intraerythrocytic levels of ATP and GTP significantly improve oxygen transfer in fish during environmental hypoxia and anemia. In contrast to environmental hypoxia, intraerythrocytic levels of ATP and GTP increase during anemia in fish facilitating oxygen unloading to the tissues. It is suggested that the increase in ATP and GTP levels during anemia occurs because the conditions required to increase the activity of adenylate and guanylate phosphate synthetic pathways are similar. PMID- 8728839 TI - Oxygenation properties of hemoglobin from the turtle Geochelone carbonaria. AB - The oxygen-binding properties of hemoglobin (Hb) from the adult terrestrial turtle Geochelone carbonaria are described. Turtle hemoglobins have a low intrinsic oxygen affinity and a low sensitivity to an endogenous cofactor (ATP) usually present at high concentrations in the reptile erythrocytes. The amplitude of the Bohr effect for O2 binding was virtually the same in the absence and presence of saturating ATP concentrations (delta logP50/delta pH, about -0.60) and increased in the total hemolysate (-0.83). The large Bohr effect found in G. carbonaria Hb may be important for O2 delivery to the tissue. The degree of cooperativity displayed by Hb for O2 binding ranged between 1.5 and 2.0 in stripped solution and total hemolysate. These observations suggest that stability of the low affinity conformation, which needs to be confirmed by additional experiments. PMID- 8728840 TI - Blood acidification in the swimbladder: mechanisms and metabolic pathways. AB - The metabolism of gas gland cells of the swimbladder epithelium is specialized for the production of acidic metabolites that are released into the blood stream and provoke an increase in gas partial pressure by reducing the effective gas carrying capacity of the blood. In a subsequent step this initial increase in gas partial pressure is multiplied by back-diffusion of gas molecules from the venous to the arterial side in the countercurrent system, the rete mirabile. Thus, gas partial pressures of up to several hundred atmospheres can be generated in the swimbladder. Measurements of metabolic end products and analysis of the formation of 14CO2 from [1-14C]glucose and [6-14C]glucose revealed that the acidic metabolites are lactic acid, produced in the glycolytic pathway, and also CO2, formed in the pentose phosphate shunt. CO2 easily enters the blood stream by diffusion. The release of protons from isolated gas gland cells, however, is highly dependent on the extracellular sodium concentration. This sodium dependence can in part be blocked by addition of amiloride, indicating that a Na+/H+ exchanger is involved in the release of protons. A significant decrease in the rate of proton secretion in the presence of the carbonic anhydrase inhibitor ethoxzolamide indicates that the second major route for the release of protons includes carbonic anhydrase activity and the diffusion of CO2. PMID- 8728841 TI - Physiological constraints in the aerobic performance of hummingbirds. AB - Hovering flight has been described as the most energetically expensive form of locomotion. Among the vertebrates, hummingbirds weighing only 1.5-20 g are the elite practitioners of this aerial art. Their flight muscles are, therefore, the most oxygen demanding locomotor muscles per unit tissue mass of all vertebrates. Tissue level functional and structural adaptations for oxygen transport are compared between hummingbirds and mammals in this paper. Hummingbirds present extreme structural adaptations in their flight muscles. Mitochondrial densities greater than 30% are observed in their pectoral muscles, and the surface area of the inner membrane of their mitochondria is twice that of mammals. This doubling of their mitochondrial oxidative capacity is accompanied by a proportional increase in the specific activity (per g tissue) of the mitochondrial manganese superoxide dismutase (SOD-Mn) in their flight muscles, thus indicating that oxygen toxicity is not a constraint in the aerobic performance of hummingbirds during hovering flight. Finally, the liver appears to play a major role in providing the necessary substrates for their high aerobic performance, and also in eliminating the oxygen free radicals formed during oxidative phosphorylation. PMID- 8728842 TI - The role of CO2/pH chemoreceptors in ventilatory control. AB - It now appears that at least some members of all classes of vertebrates exhibit ventilatory responses to changes in CO2/pH per se, including fishes. With the transition from aquatic to aerial respiration, there is an increase in the sensitivity of animals to this complex of stimuli, an increase in the variety of putative receptors possibly involved in eliciting ventilatory responses and an increase in the relative importance of this complex of stimuli in the genesis of resting ventilation. The variety of CO2-sensitive chemoreceptors present in air breathing lower vertebrates adds considerable complexity to experimental studies of ventilatory responses to CO2/pH. Because of the locations, discharge characteristics and reflex effects of the different receptor groups, most air breathing lower vertebrates show different responses to increases in CO2/[H+] due to cerebral ischemia, anoxia, metabolic acidosis and environmental hypercarbia. In some cases the differences are only quantitative, while in other cases the responses are qualitatively very different. These differences appear to reflect differences in the relative strength of the reflexes elicited by the various receptor groups and the net sum of their modulating influences when CO2/pH are altered via different routes. Although the situation is simpler in the higher vertebrates, in all cases the input from all of the CO2/[H+]-sensitive receptors appears to act as a biasing input which summates with other afferent information to modulate respiratory motor output, even in those species that breathe intermittently. PMID- 8728843 TI - Regulation of acid-base status in ectothermic vertebrates: the consequences for oxygen pressures in lung gas and arterial blood. AB - Extensive literature reports a negative delta pHa/delta t in ectothermic vertebrates, but data are scarce as to its consequences for O2 transport. In reptiles, the negative delta pHa/delta t results from an elevated lung gas PCO2 (PACO2) at higher temperatures, implying a corresponding fall of PAO2. In parallel, arterial PO2 rises with temperature, due to a combination of central vascular shunt and decreasing Hb.O2 affinity. As a result, the PO2 gradient between lung gas and blood (PA-aO2) becomes reduced at higher temperatures. In amphibians, the negative delta pHa/delta t results from combined cutaneous and pulmonary CO2 elimination. We propose that this leads to a rather temperature independent lung gas PO2. Moreover, our calculations suggest that resting reptiles and amphibians maintain a relatively large PA-aO2 also at high temperatures. The negative delta pHa/delta t in teleost fish is generally considered to be a result of modulated plasma [HCO3-]. Recent data from our laboratory suggest that acute pH adjustments at high temperatures may involve alterations of PaCO2 through gill ventilation, leading to a decrease of PaO2 with rising temperature. PMID- 8728844 TI - The coronary circulation: a fish perspective. AB - All elasmobranchs and about one-third of the teleosts studied have a well developed coronary system. In species with a coronary system, there is a large inter-species variation in the proportion of the myocardium that is supplied, with a correlation between activity level and dependence on a coronary system. In fish, most of the knowledge about the control of the coronary circulation stems from in situ or in vitro work with only a few recent in vivo studies. In salmonids, about one percent of cardiac output is returned to compact myocardium through the coronary circulation. It has been shown that, as in mammals, the coronary circulation is compromised by the mechanical work of the heart and peak coronary flow occurs during diastole. The control systems involved in the regulation of the coronary vascular resistance changes include both neuronal and humoral components. As in mammals, there are alpha- and beta-adrenoceptors located in the coronary circulation and a cholinergically mediated vasodilatation has been described. ATP, ADP and prostaglandins are probably also involved in the control of the vascular resistance, and there is evidence that local metabolites also have a strong influence on the blood flow through the coronary system. PMID- 8728845 TI - Control of heart rate during exercise in health and disease. AB - The authors present a review of their contributions over the last decade to the study of the autonomic control of heart rate during dynamic exercise under physiological and pathological conditions. These studies included the development of new methods for the evaluation of autonomic control of heart rate during dynamic exercise in man. Pharmacological blockade of sympathetic (propranolol) and parasympathetic (atropine) efferent nerves was used to demonstrate differences in time constants and power-dependent relative participation of each division of the autonomic nervous system, as predominant mechanisms responsible for the tachycardia occurring during dynamic exercise. These findings have permitted the use of properly standardized dynamic exercise (discontinuous protocol: step powers, seated position on a bicycle ergometer, 4-min duration), as a simple and noninvasive test for the evaluation of autonomic control of the sinus node. This test has proved to be useful for detecting physiological autonomic adaptations induced by aerobic training, as well as dysfunctions occurring in pathologic conditions such as Chagas' disease and hyperthyroidism. PMID- 8728846 TI - The interaction between O2 and CO2 movements during aerobic exercise in fish. AB - In resting rainbow trout, 99% of the total CO2 excreted across the gills consisted of bicarbonate (HCO3-) dehydrated to CO2 in the red cell. This value decreased to 93% during exercise, the remainder being excreted as molecular CO2 which existed in the pre-branchial blood. HCO3- dehydration consumes a proton which can be supplied from hemoglobin (Hb) through its buffer capacity or the Haldane effect. Bohr proton release from Hb upon oxygenation is maximal between 50 and 100% of Hb saturation, consistent with observations made in tench blood. At low swimming speeds, when venous blood O2 content (CvO2) was high, there was a small but insignificant acidosis as blood passed through the gills, indicating a greater release of protons than could be consumed by HCO3-. At higher swimming speeds, when CvO2 was low, there was a significant alkalosis in the arterial blood relative to the venous blood, indicating that fewer protons were released upon oxygenation than HCO3- ions were dehydrated to CO2. The disproportionate release of Bohr protons over the range of Hb-O2 saturation in the blood at the gills limits HCO3- dehydration during greater work loads, conserving the HCO3- buffer capacity of the blood and tissues. PMID- 8728847 TI - Interactions between body temperature regulation and blood acid-base status in anuran amphibians. AB - In vertebrate evolution, the transition from aquatic to terrestrial mode of life was associated with considerable changes in the respiratory system and CO2/pH sensitive receptors became fundamental. The present review focuses on the combined effects of hypercapnia and body temperature in anuran amphibians, that represent a key group for the transition. Recent studies have indicated that temperature affects the hypercapnic drive to breathe. Conversely, hypercapnia modulates the range of preferred body temperature of amphibians and central (CO2/pH) receptors are likely to be involved. PMID- 8728848 TI - Is the short-term modulation of heart rate in teleost fish physiologically significant? Assessment by spectral analysis techniques. AB - Heart rate is an important physiological variable in the control of cardiac output, even in fishes, where the importance of stroke volume has been overemphasized. Except for the myxinoids, the fish heart is innervated by cranial nerve X and the nature of this innervation is mainly inhibitory by parasympathetic fibers, although a sympathetic contribution has also been demonstrated. In mammals, cardiac innervation is not only responsible for the control of mean heart rate but it also modulates the beat-to-beat heart rate changes. These beat-to-beat changes are known as heart rate variability (HRV) and appear to be related to fluctuations in respiration and blood pressure. In this paper we demonstrate the link between cardiac innervation and HRV in several species of teleosts because HRV is greatly decreased after vagotomy or atropinization. In contrast, after abolishing the sympathetic influence with propranolol, only slight changes in total HRV are observed, indicating the restricted importance of the adrenergic innervation in determining phasic changes in HRV despite the significant tonic effect which has been demonstrated. Thus, it appears unlikely that the sympathetic influence will be present in any measured spectral component as suggested previously. Furthermore, clear spectral patterns do not always exist and this may be due to the erratic influence of respiration which is clearly faster than heart rate in all fish species studied. This differs from the slow ventilation frequency displayed by many mammalian species that exerts an influence on a beat-to-beat basis (respiratory sinus arrhythmia). Spectral patterns could also be affected by changing levels of circulating catecholamines, although this is still unproved. PMID- 8728849 TI - Neuropeptide control of the cardiovascular system in fish and reptiles. AB - Accumulating evidence shows the involvement of neuropeptides in cardiovascular control in mammals as well as non-mammalian species. Our own immunohistochemical studies indicate a sparse innervation only in cyclostomes, holostean fish and lungfish, a more extensive variation and distribution in elasmobranchs and teleosts, and a rich and varied innervation of the cardiovascular system in crocodiles and lizards. Vasoactive intestinal polypeptide (VIP), neuropeptie Y (NPY), gastrin releasing peptide (GRP) and tachykinins are present in most vertebrate groups. VIP is vasodilatory in the Atlantic cod (Gadus morhua) as in most mammalian species, but increases gut vascular resistance in the spiny dogfish (Squalus acanthias). NPY potentiates the effect of noradrenaline on skate (Raja rhina) coronary vessels, suggesting an interaction between adrenergic mechanisms and NPY early in evolution, but studies in the spiny dogfish and the crocodile also demonstrate different mechanisms for the action of NPY and adrenaline in some species. Bombesin/GRP increases flow to the gut in the spiny dogfish by an increase in somatic vascular resistance, while visceral resistance remains unchanged. In the caiman (Caiman crocodylus crocodylus) bombesin causes a shunting of blood from the lung to the gut. Substance P and other tachykinins in general increase flow to the gut, and on some occasions also increase somatic blood flow. Flow in the anastomosis of the crocodile (Crocodylus porosus) gut is increased by substance P. The results presented here are a review of several published and unpublished studies. PMID- 8728850 TI - Serotonergic vasomotor control in fish gills. AB - Serotonin (5-hydroxytryptamine,5-HT)-containing neuroepithelial cells (NECs) have been discovered in the gills of fish belonging to different phylogenetic groups, and an additional serotonergic innervation of the gill filaments is present in teleosts. The most apparent effect of serotonin is a branchial vasoconstriction. For teleosts, it has been postulated that the serotonergic innervation of the proximal portion of the efferent filamental artery (EFA) and adjacent efferent lamellar arterioles (ELA) acts to constrict the vasculature. Thus, as perfusion pressure increases, more lamellae are recruited. In contrast, recent results from rainbow trout show that one precise action of serotonin is a vasoconstriction on the distal portion of the filament vasculature. This may explain why exogenously added serotonin impairs gas exchange in rainbow trout. In addition to constricting the respiratory (arterio-arterial) pathway, injection of serotonin into the Atlantic cod dilates the arterio-venous pathway, diverting blood flow to the arterio-venous pathway. The vasoconstrictory effects of serotonin can be blocked by the 5-HT-receptor antagonist methysergide, whereas the vasodilatory effects cannot. PMID- 8728851 TI - Cardiovascular effects of adrenomedullin in teleost fishes. AB - Adrenomedullin is a fifty-two-amino acid polypeptide that was first discovered in pheochromocytoma cells, and later in the normal adrenal medullae, lungs, kidneys, and blood. In mammals, adrenomedullin has vasodepressive effects, mainly by decreasing peripheral vascular resistance. I investigated the effects of adrenomedullin in fish to see if this novel neuropeptide would have an effect in lower vertebrates, or if its actions were limited to the higher vertebrates. Bolus injections of adrenomedullin resulted in a reduction of heart rate and dorsal aortic pressure in the rainbow trout Oncorhynchus mykiss. However, adrenomedullin had no effect in the Atlantic cod, Gadus morhua. The effects of adrenomedullin in trout appear to be due to a direct action on the peripheral vasculature, as pre-treatment of celiac artery strips with tetrodoxin had no effect on the ability of adrenomedullin to relax the strip. PMID- 8728852 TI - Blood pressure control by the renin-angiotensin system and its interaction with the sympathetic nervous system in fish. AB - The renin-angiotensin system (RAS) is from an evolutionary viewpoint a well preserved regulatory system with many morphological and physiological similarities in all vertebrate groups from fish to mammals. Regulation by the RAS is often complicated, involving several other control systems and blood pressure is just one of many parameters controlled by this system. Control of cardiovascular functions by the RAS in fish has received some attention and both a direct action and modulation of adrenergic nerves by angiotensin II have been reported in most species studied. The interaction between the RAS and the adrenergic system in fish is not fully understood, but the facilitation of catecholamine release by angiotensin II is probably one of the more important mechanisms. PMID- 8728853 TI - Anoxia and ischemia tolerance in turtle hearts. AB - It has been known since ancient times that turtle hearts exhibit extraordinary tolerance to anoxia or ischemia. The mechanisms by which they accomplish this remain obscure. The most important adaptation in anoxic turtles is a rapid and dramatic decrease in metabolic rate. Nuclear magnetic resonance measurements indicate that painted turtle (Chrysemys picta) hearts respond to anoxia with a rapid decrease in phosphocreatine (PCr; to 50% of control) after which PCr remains constant for at least 4 h. ATP is defended and does not decrease while intracellular pH (pHi) decreases by 0.2 pH units early in anoxia and is then maintained constant. Softshelled turtles (Trionyx spinifer) have been demonstrated to be far more sensitive than painted turtles to anoxia in vivo. However, isolated hearts from softshelled turtles appear to be as anoxia tolerant as those of Chrysemys. During ischemia there is also little difference in cardic performance, high energy phosphates, or pHi between these two species. A peculiar feature of turtle hearts is an extremely high concentration of phosphodiesters (PDE). The role of cytosolic PDEs remains controversial but they may function as lysophospholipase inhibitors and thereby limit phospholipid turnover (Burt CT and Ribolow H, Comparative Biochemistry and Physiology, 108B: 11-20, 1994). Whether PDEs promote anoxia/ischemia tolerance is unknown but these stresses can result in membrane lipid dysfunction in mammals. Metabolic control, acid-base, and phospholipid homeostasis all play a role in anoxia and ischemia tolerance in turtle hearts. These physiologic processes are interdependent, and how they interact in these animals is unknown, but they are experimentally accessible by modern analytical methods. PMID- 8728854 TI - Estivation in South American amphibians and reptiles. AB - A number of amphibians and reptiles have cyclic behavior, becoming inactive with the coming of the dry season. In South America this pattern of activity is common, particularly in savannah-like vegetation. During the dry season amphibians burrow into the mud or soil, and either form a cocoon or increase the osmotic concentration of body fluids to reduce evaporative water loss. Some phyllomedusid tree frogs coat their body surface with skin secretion and excrete uric acid to minimize water loss. Reptiles also retreat into shelter deep enough to avoid temperature fluctuation during estivation or reduce metabolic response to temperature. Reduction of temperature sensitivity of the metabolism seems to be a strategy common to estivating amphibians and reptiles. Despite seasonal change of the environment, some species of reptiles are active all year round. PMID- 8728855 TI - Oxygen as a modulator of body temperature. AB - Most animals respond to a shortage of oxygen by lowering their body temperature. This response, mediated by behavior and physiological means, reduces oxygen demand via the Q10 effect, and should therefore be adaptive. This article reviews the occurrence of this response within the animal kingdom, the possible mechanisms and mediators of the response, and the physiological significance of hypoxia-induced hypothermia. PMID- 8728856 TI - Biochemical adjustments to hypoxia by Amazon cichlids. AB - The isozyme distribution of cichlid lactate dehydrogenase (LDH) is related to species environmental preferences. Cichlasoma amazonarum occurs in different environments and presents LDH tissue distribution patterns that correlate with oxygen tension at the capture location. Cichlasoma amazonarum was exposed to long term severe hypoxia (51 days at 36.4 +/- 5.9 mmHg), tissue LDH isozyme distribution was analyzed by electrophoresis and enzyme activities were measured by monitoring the oxidation of NADH as pyruvate was reduced to lactate. The exposure of Cichlasoma amazonarum to long-term severe hypoxia resulted in changes in the tissue distribution of LDH isozymes. The major changes in response to hypoxia occurred in heart, liver and brain: isozyme A4 was activated in heart and brain, whereas isozyme B4 was activated in liver. The most significant quantitative change occurred in brain LDH of hypoxia-exposed animals which adopted muscle type kinetics, reflecting a new LDH isozyme distribution. LDH activity was significantly reduced (P < 0.05) in animals exposed to hypoxia (N = 8), suggesting an overall LDH suppression. Pyruvate inhibition decreased in all hypoxia-exposed tissues. Thus, the ability of Cichlasoma amazonarum to regulate LDH tissue expression according to oxygen availability allows the animal to survive chronic hypoxic environments. This phenotypic plasticity may occur in other hypoxia-tolerant fish species. PMID- 8728857 TI - The heart of the icefish: bioconstruction and adaptation. AB - The Channichthyidae or "icefish" represent an intriguing example of extreme adaptation to the stable low temperature and high oxygen content of the Antarctic waters. The lack of respiratory pigments (hemoglobin and myoglobin) in these teleosts is associated with relatively low oxygen consumption and relevant cardio circulatory adjustments which include large blood volume, increased relative heart weight (cardiomegaly), and very high cardiac output. The heart has the ability to displace large systolic volumes at a low rate and relatively low pressure, with large ventricular fillings (high ventricular compliance), whereas it is incapable of facing increased afterloads. These functional aspects of mechanical flexibility and restrictions of the cardiac pump have been tentatively related to some constructional aspects of the icefish cardiomegaly, particularly, at the whole ventricular level, to the trabeculate type of myo-architecture, and, at the subcellular level, to the conflict in space economy between the exceptionally high mitochondrial densities and the consequent severe reduction in myofibrillar volume. On the basis of this morphodynamic approach, we suggest that the icefish may illustrate how a certain feature (i.e., an architectural cardiac design) common to the suborder and to most teleosts, and apparently with "irrelevant" properties, can become useful for a specialized purpose (i.e., volume pump design); and how, in contrast, the internal machinery construction, because of structural and ultrastructural constraints, may prevent these stenothermal sedentary teleosts from conquering niches requiring more active locomotory habits. PMID- 8728858 TI - Electrocardiographic characterization of myocardial function in normoxic and hypoxic teleosts. AB - Electrocardiography was applied to analyze cardiac function of four teleost species (Piaractus mesopotamicus, Hoplias malabaricus, Hoplias lacerdae and Cyprinus carpio) during normoxia and graded hypoxia. In these species, hypoxic bradycardia consistently occurred during severe hypoxia (below the critical oxygen tension--PCO2) and was accompanied by alterations in the ECG recordings. Three basic ECG alterations were demonstrable: 1) increase in the T wave area and amplitude, being more positive and with symmetrical morphology during severe hypoxia (P. mesopotamicus); 2) negative T wave in normoxia, changing to isodiphasic (just above the PCO2) and positive (below the PCO2; H. malabaricus and H. lacerdae); 3) positive T wave in normoxia, changing to negative in severe hypoxia (5 mmHg; Cyprinus carpio). These findings indicate changes in the direction of ventricular repolarization during exposure to severe hypoxia, and the analysis of the ECGs in relation to the derivation line permitted the estimation of these drifts to be 17 degrees in P. mesopotamicus, 46 degrees in H. malabaricus, 43 degrees in H. lacerdae, and 32 degrees in C. carpio. The changes in the direction of ventricular repolarization were attributed to myocardial impairment due to insufficient oxygen supply, and support the idea of a relationship between cardiac dysfunction and the bradycardia developed during severe hypoxia. PMID- 8728859 TI - Cardiovascular measurements in animals in the milligram range. AB - The study of microscopic animals should be intensified because: most of the world's animal biomass consists of very small animals; life as a small animal is both qualitatively and quantitatively very different from that of a large animal; and almost all animals are very small as they begin their development. Fortunately, developing technology now allows us to make quantitative measurements in microscopic animals. This paper describes new techniques for measuring cardiovascular variables such as blood pressure, stroke volume, heart rate and cardiac output in animals weighing as little as a few mg. Non-invasive techniques such as videomicroscopy can be used for determining heart stroke volume in small animals. Impedance measurement is another non-invasive or minor invasive technique for determining rates of heart beat, gill or lung ventilation and limb movement as well as giving qualitative information on changes in blood flow. Pulsed Doppler technology can be used to obtain blood flow velocity in small vessels. Invasive techniques depend on servo-null micropressure systems that record pressure through glass microelectrodes that are implanted into the vessel or heart lumen. This allows stable pressure recordings for up to 5-6 h in animals weighing as little as a few mg. Microinjectors can be used for intravascular injections of vasoactive drugs (or blood withdrawals). Newly emerging techniques for in vivo cardiovascular measurements allow us to understand the function of the cardiovascular system in a larger portion of the world's animal biomass, as well as in the immature and as yet poorly understood early developmental stages of animals. PMID- 8728860 TI - The use of data loggers to determine the energetics and physiology of aquatic birds and mammals. AB - By deploying a data logger specifically designed for the purpose, it was possible to record heart rate, fH (beats/min), from free-ranging gentoo penguins, Pygoscelis papua, and Antarctic fur seals, Arctocephalus gazella, at the British Antarctic Survey Base at Bird Island. The heart rate data were then converted into oxygen consumption (VO2, mlO2 min-1 kg-1) and/or energy expenditure (W/kg) using equations that had been derived from calibration experiments. In these experiments the relationships between fH and VO2 were determined in animals at rest and while exercising at different levels on a treadmill or in a static water channel (penguins) and in a variable speed flume (California sea lions, Zalophus californianus, as surrogate fur seals) or in a static water channel (fur seals). The validity of using these relationships was tested by recording simultaneously fH, VO2 by direct respirometry and VCO2 by the doubly labelled water (DLW) technique in six penguins and in six California sea lions during 72 h and 24 h, respectively, at various levels of activity. For the penguins, both indirect methods gave mean algebraic errors within 2% of the measured VO2, whereas for the sea lions, the mean algebraic errors were 36.4% for the DLW method and 2.7% for the fH method. The range of errors was greater for the DLW method, in both species. Field data from 15 penguins indicate that the fH method provides data that are comparable to those obtained using the DLW method, but with the added advantage that they can be broken down into the energy costs for specific types of behaviour. The implanted data loggers also recorded the temperature of the abdominal cavity (T(ab)) and it was evident that this routinely decreased by approximately 2 degrees C during diving bouts and by a maximum of almost 5 degrees C. Such temperature decreases, particularly if representative of similar decreases in other tissues, may at least partly explain why the energy costs of travelling to and from the foraging site and of foraging itself are similar to those for penguins resting in water at 5 degrees C. Field data from 15 female fur seals indicate that, when the animals are ashore, there is good agreement between the values for mean energy expenditure obtained by the fH and DLW methods. However, when the animals are at sea, the values obtained by DLW are substantially greater than those obtained by fH. When the at-sea values are corrected for the apparent overestimations referred to above, there is good agreement between the data obtained using the two methods. The data derived from fH indicate that, as with the penguins, the energy expenditures during travelling to and from the foraging site and during foraging are similar to those of fur seals resting in water at 7 degrees C. PMID- 8728861 TI - K+ balance in rainbow trout gill and liver tissue, cell suspensions and cultured cells. AB - Both intact gill and liver tissue from rainbow trout accumulated K+, as determined by 86Rb+ uptake, a process largely inhibited by ouabain, indicating the presence of functional NaKATPase. Cell suspensions, produced by disaggregation of gill or liver tissues, accumulated very little K+ compared to tissues (less than 10%). Disaggregation resulted in depolarisation of cells with loss of intracellular K+ and although NaKATPase, as measured by 86Rb+ uptake rate, remained functional and inhibitable by ouabain, the activity was insufficient to replace the rapid K+ loss. While attached, cultured gill and liver cells showed normal K+/Na+ ratios and NaKATPase activity, but release from the substratum resulted in depolarisation and rapid K+ loss as seen in cell suspensions. These results suggest that care is required in interpreting ionic regulatory and other results from cell suspensions and that further research should be directed towards systems where cells can maintain normal ionic balance. PMID- 8728862 TI - Magnetocardiography in normal subjects and in patients with right bundle branch block. AB - The initial results of a magnetocardiographic study comparing normal subjects to patients with right bundle branch block due to Chagas' and other diseases is presented. A quantitative criterion to discriminate normal healthy individuals from patients was proposed based on the amplitude of R and S waves detected by the magnetocardiogram. PMID- 8728863 TI - Fetal heart rate monitoring by magnetocardiograms. AB - The magnetic field produced by the fetal heart magnetocardiogram (FMCG) was measured by a single channel SQUID biogradiometer in an unshielded environment. FMCG amplitude ranged from 1 to 4 pT. Instantaneous fetal heart rate and averaged FMCG waveforms were computed. Spectral analysis was used to quantify fetal heart rate variability, and revealed peaks from 0.5 to 1.0 Hz, consistent with respiratory sinus arrhythmia. The duration of the QRS complex was approximately 50 msec, and its amplitude and shape showed strong spatial variation. Signal-to noise ratio was inadequate for the study of P and T waves. These data show that this relatively simple and inexpensive instrument can be used for noninvasive in utero investigation of fetal heart activity without requiring a special environment. PMID- 8728864 TI - The frog brainstem preparation as a model for studying the central control of breathing in tetrapods. AB - Our frog brainstem preparation revealed mechanisms for the central control of breathing that are in many ways similar to those of mammals. Thus, the basic control mechanisms for air-breathing appear to have been present in the Devonian common ancestors of frogs and mammals and may be common to all lung-breathing vertebrates. LOCATION: The in vitro frog brainstem, including motor nuclei of cranial nerves V to X, maintains frequency and ratio of fictive buccal oscillations to fictive lung inflation episodes comparable with that of the living animal. In this preparation, transection caudal to V abolishes spontaneous discharge in X but slow, spontaneous discharge in V may remain. Independent central pattern generation is present in the left and right half-brainstems. CHEMOSENSITIVITY: The frequency of fictive lung inflations increases with decrease in pH within the physiological range. RESPONSE TO GLUTAMATE: Biphasic response, consisting of a pause, followed by a dramatic increase in the frequency of fictive inspirations and positive baseline deflection, followed, in turn, by slow return of the baseline to the control level with frequency remaining above control as long as glutamate is applied. Local application reveals glutamate sensitive sites in the ventral reticular formation. RESPONSE TO SUBSTANCE P AND PHYSALAEMIN: Similar to glutamate but the frequency of fictive inspirations decreases below control values. RESPONSE TO STRYCHNINE: The normal temporal sequence in firing of motor neurons of cranial nerves is disrupted and all nerves are synchronously active. The firing sequence of respiratory neurons is consistent with a grouping possibly homologous to the mammalian inspiratory, post inspiratory and expiratory phases. PMID- 8728865 TI - Methods for studying respiratory mucus and mucus clearance. AB - We describe some basic procedures for studying the properties of mucus. These techniques can be applied in both clinical and physiological studies to improve the understanding of the mechanisms related to epithelial defense in health and disease. Mucus collection--A major difficulty is the lack of simple and noninvasive methods for collecting normal mucus in sufficient quantity for later analysis. Physical properties of mucus--A. Rheology: Mucus exhibits both solid and liquid properties and the important factor governing the actual behavior is time. The magnetic microrheometer provides an elegant method for measuring rheological properties of microsamples. B. Adhesivity: It characterizes the forces of attraction between an adherent surface and an adhesive system and can be calculated by measuring the contact angle between a mucus drop and a surface. Mucus Transport--A. Mucus transport by cilia: Mucus is primarily cleared by the continuous ciliary beating, which can be studied using techniques such as the frog palate preparation as well as direct measurement, i.e., in situ mucus clearance. B. Cough clearance: It is essential for elimination of secretions in diseases leading to hypersecretory states. The cough machine simulates the flow time profile of human coughing. Transepithelial potential difference--A potential difference exists between the epithelial surface and the submucosa and is the net result of the activity of the ion-transport system of the pulmonary epithelium. The potential can be measured using appropriate microelectrodes. Quantitative morphology--Methods may be used to characterize the epithelial surface condition that continuously changes during aggressive conditions. PMID- 8728866 TI - [Cold-induced cardiac abnormalities in systemic sclerosis]. AB - Cardiac abnormalities are frequent in patients with systemic sclerosis (SS). These abnormalities have been demonstrated in over 80% of patients with SS and there are some clues that suggest that an intermittent vascular spasm (i.e. coronary Raynaud's phenomenon) is one of the causes of myocardial dysfunction in this group of patients. The aim of this prospective study was to evaluate the ventricular performance and regional wall motion during exposure to cold in patients with SS and Raynaud's phenomenon without overt cardiac disease. Twenty four patients and 10 normal volunteers underwent radionuclide ventriculograms (RV). In each subject the RV was done thrice: basal, 20 minutes after chilling with thermic blanket and post reheating. The left ventricular ejection fraction (LVEF) decreased during the cold test (p = 0.03) with reversible abnormalities in wall motion. Basal Right Ventricular Ejection Fraction (RVEF) was lower than that of normal subjects (p = 0.02) and decreased during the cold test (p = 0.04). Therefore, we were able to demonstrate an early cardiac dysfunction associated with impaired wall motion after corporal chilling. These findings suggest that coronary spasm in SS would be an early and frequent phenomenon that would precede the development of symptomatic cardiac disease in some patients with this illness. PMID- 8728867 TI - Evaluation of HTLV-I/II infection in blood donors in Buenos Aires. AB - HTLV-I and HTLV-II are two related retroviruses that are transmitted by sexual contact, breast feeding, blood transfusion and needle sharing. In this study the prevalence of HTLV-I and HTLV-II was evaluated in voluntary blood donors as a measure of the infection in the general population. Samples were tested by a gelatine particle agglutination test and repeatedly reactive samples were confirmed by Western blot tests (WBT), enriched with recombinant rgp21, rgp461 y rgp4611 proteins, which differentiates HTLV-I and HTLV-II antibodies. Of 19,426 samples, 40 were repeatedly reactive by particle agglutination (0.21%). When analyzed by WBT, 6 met the criteria for HTLV-I (0.036%), 2 for HTLV-II (0.01%) and 1 for HTLV-I/II, 13 samples were indeterminate and 18 were negative. The prevalence is low and comparable to that from non endemic countries. Screening for anti HTLV-I/II antibodies is necessary to prevent transmission through blood transfusions. PMID- 8728868 TI - [Gas exchange changes in the postoperative period of heart surgery]. AB - We studied prospectively 59 open-heart surgical patients (CBPS) in order to evaluate postoperative arterial blood gases evolution and its predictive value of respiratory and non-respiratory post-surgical complications. Twenty-four hours after CPBS 28 over 59 patients showed left pleural effusion and/or left lower lobe atelectasis. 62.9% or pleural effusions were only blunted costophrenic angle. Chest x-ray film were normal in 38.9% (23/59) of patients. Forty-eight hours after CPBS only 5% (3/59) radiographs were normal and only 31% of pleural effusion were classified as minimal. Forty-eight hour radiographs worsened in 69.4% of the patients. (Table 1). During 48 hours period 71% of patients showed pleural effusion and 42% atelectasis. Only 1 patient showed an atelectasis up a third of hemithorax (3.5% of abnormal chest X-ray films). Twelve hours alveolo arterial quotient (a/A) was decreased in 50/59 patients (0.51 +/- 0.16), more deeply at the second day. There was no relationship between CBP time and a/A at 12 or 48 hours. The normal chest X-ray film patients mean a/A was no different (0.54 +/- 0.17). The a/A at 48 hours was no different between patients with and without lower lobe atelectasis. Nine patients (15%) developed respiratory complications (RC) and 11 (19%) non-respiratory complications (NRC) (Table 2). There was no difference in CBP time (76.9 +/- 27.9 vs 88.1 +/- 27.7 min p = NS) nor aortic cross-clamp time (52.61 +/- 20.43 vs 59.57 +/- 19.39 min p = NS) between patients with and without RC. There were no differences in a/A at 12 hours (0.47, 0.51, 0.48 p = NS) and 48 hours (0.34, 0.32, 0.30 p = NS) between patients without complications, with RC and with NRC (Table 3). There was no correlation between 12 or 48 hours a/A and intensive care or hospital stay length. The absence of predictive value of hypoxemia could be explained from a different source of early a/A fall and important RC. It could mean that RC after CPBS are not specific of that sort of surgery nor involve mechanisms related to that special intraoperative circumstances, which is not the same for gas exchange alterations. We conclude that a/A deterioration is a very common finding after CBP and does not identify particularly risky patients. PMID- 8728869 TI - [In vivo measurement of the mineral content of renal calculi by dual-photon absorptiometry. Correlation with its fragility to extracorporeal shockwave lithotripsy]. AB - After a few years of experience with extracorporeal shock wave lithotripsy (ESWL) and other fragmentation techniques, it has become apparent that stone fragility is a significant clinical distinction that should be taken into consideration when selecting a treatment program. In 30 unselected patients, stone mineral content, density and area were measured in vivo by dual-photon absorptiometry prior to perform ESWL treatment. Stone area determinations showed a median of 4.21 with a range of 0.46 to 49.7 cm2. Stone mineral content (g) and stone density (g/cm2) values were 2.47 and 0.46 with ranges of 0.37 to 13.7 and 0.167 to 1.203 respectively. The number of shocks needed for total fragmentation were 2375 with a range of 1200 to 7800. No correlation could be found between the number of shocks needed for fragmentation and the stone area or density. On the other hand, a strong linear correlation (r = 0.81, p < 0.001) (Fig. 1) could be demonstrated between stone mineral content and the number of shocks needed for fragmentation. Our results support the concept that size alone is not always a suitable criterion for selecting a stone as appropiate for ESWL, since no correlation could be found between stone area and the number of shocks needed for total fragmentation. We were also unable to find any correlation between in vivo stone density measured by dual-photon absorptiometry and the number of shocks required for stone fragmentation. Instead, a strong linear correlation between stone mineral content and its resistance to shock wave fragmentation was found. Therefore, calculation of mineral content appears to be the determinant of the amount of energy required for total fragmentation. Our results strongly suggest that in vivo stone mineral content measurement provides helpful information for predicting the fragmentation prospect of a stone. PMID- 8728870 TI - Blood sugar concentrations during ketamine or pentobarbitone anesthesia in rats with or without alpha and beta adrenergic blockade. AB - Plasma glucose concentrations were measured in: 1) conscious and anesthetized rats during an iv glucose tolerance test (IVGTT) and 2) conscious and anaesthetized phentolamine/propranolol blocked rats during an IVGTT. Anesthesia was induced with ketamine (120 mg.kg-1) or pentobarbitone (60 mg.kg-1) ip at -30 min of the beginning of the IVGTT, which was followed by 2 injections of the anesthetic agents at intervals of 30 min. Propranolol (2 mg.kg-1) was given ip at -25 and -5 min. An iv infusion of phentolamine (0.015 mg.min-1) was started at 20 min and continued up to the end of the experiment. During the IVGTT, the anesthetized rats showed a moderate hyperglycemic response to glucose load compared to conscious animals (ketamine: p < 0.01 at 5 min; and p < 0.05 at 10-20 min; pentobarbitone: p < 0.05 at 5-20 min). The hyperglycemic response to glucose administration in the conscious rats was not affected by adrenergic blockade (p > 0.05). While in ketamine anesthetized rats the increased glucose response was abolished by adrenergic blockade (p < 0.05 at 5-10 min), this effect was not seen in pentobarbitone anesthetized animals. These results suggest the existence of an inhibitory tone on insulin secretion and a glycogenolytic response in ketamine anesthetized rats, probably mediated by adrenergic inervation of the pancreas and liver and by circulating catecholamines secreted from the adrenal medulla. PMID- 8728871 TI - [Low density lipoprotein rich in triglycerides and hepatic lipase activity in insulin-dependent diabetic patients]. AB - Genetic hepatic lipase (HL) deficiency is associated with low density lipoprotein (LDL) rich in triglycerides (TG), whose affinity for B:E receptors is decreased. In rats, experimental hypoinsulinemia produces HL deficiency. However, the relation between human insulin-dependent Diabetes Mellitus (IDDM), HL activity and the characteristics of LDL have not been studied. The objective of our study is to evaluate the relation between HL activity and the chemical composition of LDL in treated IDDM patients. Subjects were 15 IDDM patients and 15 controls (C), matched for sex and body mass index (BMI). The IDDM patients were classified by the WHO criteria, were free of nephropathy and hypothyroidism, and received no medication except insulin. Controls were clinically healthy and normolipidemic with no family history of diabetes. The IDDM group was divided into two subgroups: subgroup IDDM-A (n = 9) with HL values > or = 4.3 and IDDM-B (n = 6) with HL < or = than 4.2 mumoles glycerol/ml h. the HL in IDDM was lower than in C (p < 0.001). Table 1 shows clinical data. Blood samples were drawn after 12 h fasting. Percentage of HbA1c and plasma concentrations of glucose, total cholesterol, LDL-cholesterol, HDL-cholesterol and TG were assayed. LDL was separated by sequential ultracentrifugation at densities of 1.019-1.063 g/ml and its chemical composition was analyzed. The most relevant results were: plasma TG concentration was higher in IDDM than in C (p < 0.05) (Table 2), although average values DMID not exceed the reference values of 200 mg/dl. The TG-LDL were higher in IDDM than in C: 24.8 +/- 2.7 vs 17.5 +/- 1.1 mg/dl plasma, media +/- SE, (p < 0.02). This difference reflected the values of IDDM-B, whose plasma concentrations of TG-LDL were higher than in C: 32.3 +/- 3.6 vs 17.5 +/- 1.1 mg/dl (p < 0.001), and also higher than in IDDM-A (p < 0.02). (Table 3). The chemical composition of LDL in IDDM-B contained a higher percentage of TG than C: 8.5 +/- 0.7 vs 6.8 +/- 0.3% (p < 0.05), a lower percentage of cholesterol than IDDM-A: 39.0 +/- 1.7 vs 45.2 +/- 2.2% (p < 0.05) and also a larger percentage of proteins than IDDM-A: 28.9 +/- 1.9 vs 20.8 +/- 1.0% (p < 0.01). The correlations between TG/cholesterol and HL activity in IDDM were r = -0.53 (p < 0.05) and in IDDM-B, r = -0.81 (p = 0.05). The noteworthy result of this study is the modification of the LDL particle in IDDM, rich in TG in patients with low HL activity. Anomalies in the chemical composition of LDL like those described decrease the uptake of this particle by its physiological B:E receptors. It has recently been demonstrated that LDL is an indisoluble association of lipids and apoproteins, and that both act simultaneously to hold the apoB in a spatial position that expresses normal epitopes. It has been described that particles of LDL rich in TG and poor in cholesterol, shows low affinity for LDL receptors in human fibroblasts. Also in IDDM the interaction of LDL rich in TG with B:E receptors is decreased. This might be one more mechanism contributing to the accelerated atherosclerosis of these patients. Our results suggest that there may be a threshold of HL activity for the complete hydrolysis of the TG of LDL, for the normalization of the TG/cholesterol relation and for the conformation of typical LDL particles. PMID- 8728872 TI - [Chronic cough in pediatrics]. AB - The objective has been to identify the different etiologies and elaborate a diagnostic and therapeutical methodology for patients with chronic cough. During one year we studied prospectively 83 patients with persistent cough of daily appearance with an evolution of four or more weeks and no previous etiologic diagnosis. We worked on three diagnostic (D) levels. D1: Based on the anamnesis and physical examination. D2: Sequential incorporation of complementary exams. D3: Evaluation of the response to the specific treatment. We divided the population into 2 groups: G1 healthy children, G2 children followed in our hospital for different conditions. The mean age was 4.7 years (range, 3 months to 15 years), and the average duration of cough was 4.9 months (range, 1 to 36 months). In G1 the following causes were identified in 78 children: cough variant asthma 41 (52%), asthma+upper respiratory tract infections 8 (10%), asthma+lower respiratory tract infections 6 (7%), postnasal drip syndrome (sinusitis, adenoiditis) 5 (6%), psychogenic 6 (7%), undetermined 4 (5%), gastroesophageal reflux 2, asthma+cigarette 2, AIDS 1, Sjogren syndrome 1, vascular ring 1, cricopharyngeal foreign body 1. In G2 out of 5 children we have found: 2 children with chronic encephalopathies who had swallowing disorders and gastroesophageal reflux, 1 patient with Down syndrome presenting hypogammaglobulinemia and bronchiectasis, 1 tracheaesophageal fistula in H in a child with recurrent pneumonia, 1 lymphocytic pneumonia in an AIDS patient. The D1 was correct in 92% of the cases. The specific therapy has proved useful for achieving the remission of the symptoms. Although asthma is the most frequent cause of chronic cough, other etiologies exist and must be ruled out. PMID- 8728873 TI - Cellular adaptation of the rat medullary thick ascending limb to chronic renal failure. AB - Chronic renal failure (CRF) is accompanied by adaptive changes in renal and extrarrenal epithelial ionic transport. Fluid reabsorption in the thick ascending limb of Henle is increased and the capacity to lower the urine osmolality in water diuresis is preserved. To study the cellular mechanism of this adaptation, we measured intracellular cAMP in microdissected medullary thick ascending limb (mTAL) segments in rats with CRF. mTAL exhibited in CRF nephrons an increase of basal cAMP from 6.0 +/- 1.5 in controls to 47.0 +/- 10.3 fmol. mm-1 tubule in CRF (P < 0.05). Maximally stimulated cAMP levels (10(-3) M IBMX plus 10(-5) M Forskolin) were different from basal levels in controls (6.0 +/- 1.5 vs 63.1 +/- 18.8, P < 0.05) but not from basal levels in CRF (47.0 +/- 10.3 vs 63.0 +/- 16.0, P = N.S.). Preincubation with the adenylate cyclase inhibitor 2'5' dideoxyadenosine (DDA) 10(-4) M produced no changes in cAMP in controls (93.7 +/- 10.3% of DDA untreated samples) whereas it decreased to 76.2 +/- 8.8% (24% inhibition) in CRF (P < 0.05). No differences between controls and CRF groups were found in basal and stimulated cAMP in red blood cells and distal colon. The data would suggest that the cAMP pathway is an intracellular signal for mTAL adaptation in epithelial transport and that the adenylate-cyclase system is specifically activated in CRF. PMID- 8728874 TI - [Probable HIV transmission by female homosexual contact]. AB - Here we report a case of HIV infection in a lesbian lacking other risk factors. She had sought advice for asthenia and unexplained weight loss. The physical examination disclosed oropharyngeal candidiasis and generalized lymphoadenopathies. The patient defined herself as an active sexual partner and denied having any heterosexual intercourse whatsoever. Particle agglutination test proved positive for anti-HIV, as well as two consecutive ELISA assays, confirmed by Western blot. Since there was no history of blood transfusions or intravenous drug addiction, it may be concluded that HIV transmission was most likely due to female homosexual practices. PMID- 8728875 TI - [Primary bronchial actinomycosis and foreign body]. AB - This paper presents the fourth case reported on the association of primary bronchial actinomycosis and foreign body. The pathogenesis of this rare association has been linked to the low respiratory tract and a foreign body (chicken bone); it depends on its characteristics and how long it has remained lodged into the bronchial tree. The diagnosis of this case was very difficult and late. Several endoscopy procedures were required in order to detect it. Our patient has been treated successfully with parenteral penicillin in association with an original bronchoscopic procedure using a laser technique to find the foreign body and take it out of the respiratory tract. We consider that it is necessary to remember this association in every patient who suffers from recurrent pneumonia, in those who are at risk of foreign body aspiration and when the presence of an endobronchial mass is suspected. PMID- 8728876 TI - -Prolonged fever syndrome and infection of abdominal aortic aneurysm due to Salmonella enteritidis. AB - Endovascular infection of atherosclerotic aorta is a rare event in the setting of aged patients with gram negative bacteremia of the salmonella group. Until the beginning of the 60s this meant an ominous diagnosis with an almost unavoidable fatal prognosis. Presently, this trend has been reverted, mostly due to an earlier diagnosis, the development of more sophisticated imaging techniques, the correct use of broad spectrum bactericidal antibiotics and prompt surgical management. Paradoxically, the incidence of arterial infections has increased in recent years, specially in old people with atherosclerotic abdominal aortic aneurysms, in whom infective endocarditis could not be demonstrated. We describe the case of a 65 year old man, with a history of longstanding non-insulin dependent diabetes, presenting with protracted fever, weight loss and thigh pain. Blood cultures and serologic studies as well as several echocardiograms yielded negative results. An abdominal CT scan showed an infrarenal aortic aneurysm raising the clinical suspicion of arterial infection of abdominal aorta. The patient underwent surgery because of highly presumptive diagnosis of complicated aortic aneurysm. The resection was followed by an in situ graft. There was no evidence of disruption or gross collection. Samples of the aortic wall and perianeurysmatic fluid grew Salmonella enteritides. We describe the main etiopathogenic and clinic features of the entity highlighting the high sensitivity and specificity of the CT scan in the identification and characterization of infected aortic aneurysm. Certain features may firmly suggest this diagnosis without using preoperative aortography. PMID- 8728877 TI - [Micromolecular multiple myeloma, hypertension and nephropathy]. PMID- 8728878 TI - [Role of excitatory amino acids in neuropathology]. AB - Excitatory amino acids (EAA) became known as neurotransmitters of the central nervous system (CNS) in the last decade. The most studied EAA are glutamate and aspartate. Both are synthetized by the same mechanism as gamaaminobutyric acid. (Fig. 1). Glutamate is widely distributed in the CNS and the spinal cord, being the areas of higher concentration the cerebral cortex, the hypocampus and the cerebellum. There have been identified two type of receptors for glutamate: ionotropic and metabotropic. The former includes three different types: NMDA, AMPA and KA. NMDA receptor is coupled to a Na+ and Ca2+ channel being the second ion the most important one. This receptor has several sites of binding for various substances. Along with the site for N-methyl-D-aspartate, which binds glutamate and/or aspartate, there have been identified a site for the binding of glycine (which is different from the strychnine sensitive one), a site for poliamines such as spermine and spermidine, and a site for the binding of Zn2+ (Table 1). AMPA receptor is associated to a Ca(2+)-Na+ channel, being in this case the Na+ the most important ion. There are two metabotropic type receptors: L AP4 and trans-ACPD. Both are coupled to a G protein and agonists exert their action increasing phospholipase C activity which in turn induces an increment of IP3 and diacyl-glicerol, and a consecutive releasing of Ca2+ from intracellular stores. EAA play a role in some physiological processes. One of them is long-term potentiation (LTP), an electrochemical phenomenon involved in memory consolidation. Antagonists of NMDA and AMPA receptor prevent the development of LTP, and conversely, the agonist of glycine site of NMDA receptor--D-cycloserine- facilitates memory consolidation. Since 1957, EAA are considered neurotoxic substances and there are many indirect evidences to support this statement. Pathogenesis of neuronal damage elicited by EAA involves the events shown in Fig. 3. Prevention of the cascade of events that provokes neurotoxicity may be achieved by NMDA antagonists, but once it has begun it may be only aborted subtracting the Ca2+ from the medium, using nifedipine or blocking AMPA receptor with an antagonist (CNQX). EAA have been shown to play a toxic role in neuronal damage induced by ischemia. Research using various experimental models demonstrated that NMDA receptor antagonists (i.e. MK 801) blocks postischemic damage. Interventions at various levels of the pathogenic cascade shown in Fig. 4 provoke the same results. There is enough evidence to suspect that NMDA and AMPA receptors are altered in epilepsy. NMDA antagonists (i.e. MK801 or AP5) prevent the development of epileptic seizures induced by kindling; CNQX, an AMPA antagonist, blocks the increase in electrical activity induced by K+ in slices of hypocampus; felbamate, an antiepileptic drug, blocks the glycine site (not strychnine sensitive) decreasing NMDA receptor activity. Several neurodegenerative disorders have been associated with exogenous administration or accidental intake of EAA. (i.e. neurolatirism, Guam disease). Similarities between these diseases and lateral aminotrophic sclerosis indicate that in the latter EAA may play a pathogenic role. Finally, the psychotomimetic effect of phencyclidine (an antagonist of NMDA receptor) suggests that in schizophrenia, together with dopaminergic neurotransmission impairment, some dysfunction of glutamate pathways may be present. PMID- 8728879 TI - [Autologous transplantation of peripheral blood stem cells in malignant lymphomas]. AB - The administration of high dose chemotherapy and or radiotherapy with autologous hematopoietic rescue has become a treatment modality with increasing number of indications in a variety of malignant conditions. Improvements in the conditioning regimens and supportive measures used, as well as a more refined patient selection based on prognostic factors, have resulted in progressively better results. The availability of precursor cells from peripheral blood has allowed a faster restoration of hematopoiesis, decreasing the period and intensity of myelosuppression. The following revision gives an updated image of the accumulated experience with this mode of support in malignant lymphomas. PMID- 8728880 TI - [The real importance of basic research and the present status of CONICET]. PMID- 8728881 TI - [Joseph Goldberger, pellagra and prejudice]. PMID- 8728882 TI - [Natriuresis: new concepts from molecular biology]. PMID- 8728883 TI - [Acute arterial ischemia due to ergotamine simulating Takayasu disease]. PMID- 8728884 TI - [Mass screening of deafness in neonates using otoacoustic emissions]. AB - There is no clinical sign of hearing loss in the infant. Defective language acquisition and altered voice quality always occur late making early diagnosis essential for therapeutic management of deaf children. Subjective assessment of behavioral is highly dependent on the experience of the examinator and cannot be used as an effective routine method. Electrophysiological recordings of otoacoustic emissions have the advantage of allowing objective measurement. Otoacoustic emissions originate in the inner ear. A quantity of energy is transmitted via the tympanic membrane which acts as a microphone, allowing recordings in the external auditory canal. In certain infants these emissions are spontaneous and can be used to detect certain types of hearing loss. Stimulation is required however for screening tests. Recording provoked otoacoustic emissions is simple. A small probe in placed in the external auditory canal. This probe carries two microphones, one which emits clicks recorded by the other captor microphone. The examination lasts approximately 15 minutes, including the time needed to explain the procedure to the mother. There are two possible results. The presence of recorded otoacoustic emissions confirms that the inner ear is normal. If the absence of emissions is confirmed at a second examination, an evoked potential recording is required to identify the hearing loss. This safe, simple and reliable method for detecting hearing loss in infants is recommended for screening programmes during the neonatal period. PMID- 8728885 TI - [Mechanisms of diurnal hypercapnia in sleep apnea syndromes associated with morbid obesity]. AB - OBJECTIVES: The prevalence and mechanisms of diurnal hypercapnia in subjects with sleep apnea syndrome are not well known, particularly in the morbidly obese. METHODS: We studied a group of 24 subjects with sleep apnea syndrome and morbid obesity defined as a body mass index greater than 40 kg/m2 and free of chronic respiratory disease. RESULTS: Hypercapnia (PaCO2 > 42 mmHg) was found in 50% of the subjects. Age, body mass index, waist/hip ratio, apnea index, ratio of maximum forced expiratory volume in one second (FEV1) to vital capacity and expiratory reserve volume were not significantly different between hypercapnic and normocapnic subjects. Total pulmonary capacity, vital capacity and FEV1 were significantly lower in hypercapnic subjects than in normocapnic subjects. CONCLUSION: These findings suggest that ventilatory restriction plays an important role in the development of diurnal alveolar hypoventilation in subjects with sleep apnea syndrome and morbid obesity. PMID- 8728886 TI - [Prevention of hemorrhagic recurrences caused by rupture of esophageal varices, with endoscopic ligation. Prospective study of 50 patients]. AB - OBJECTIVES: Endoscopic sclerotherapy is effective to prevent bleeding of oesophageal varices but is associated with frequent adverse effects. Endoscopic ligation represents a new endoscopic alternative treatment to sclerotherapy. The purpose of this study was to assess efficacy and safety of endoscopic variceal ligation in 50 consecutive patients with cirrhosis who had recently bled from oesophageal varices. METHODS: Patients were followed from 6 to 1140 days (median 310 days). Nine patients were bleeding actively when ligation was performed. RESULTS: Eleven patients (22%) had 13 recurrent bleedings requiring blood transfusion during follow-up. Six recurrences occurred during the first month (3 from bleeding varices, 3 treatment-induced); seven recurrences occurred latter (6 from bleeding varices, 1 treatment-induced). Varices were eradicated in 33 patients (66% of all patients, 82% of patients who survived more than 30 days). Variceal eradication was achieved in 2-9 endoscopic ligation sessions (median 3). Eighteen patients died during the study; one died from bleeding. No patient developed major complications; five patients only complained of mild dysphagia for 24 to 48 hours. CONCLUSION: Endoscopic ligation is a safe and effective method to prevent recurrent bleeding from oesophageal varices. PMID- 8728887 TI - [Post-traumatic psoriatic arthritis. 2 cases]. AB - Physical trauma is generally accepted as a possible factor in the pathogenesis of rheumatoid arthritis. In the last ten years, there have been a few rare case reports of physical trauma precipitating psoriasic arthritis. We observed two such cases following an occupational accident discovered one and a half year and two and a half year after onset of the first clinical manifestations. In the first case, a 43-year-old man had a fracture of the right calcaneus in March 1991. He was treated with nailing and also required emergency surgery of the posterior tibial artery. The tibiotarsal joint was normal radiologically. Pain persisted after treatment and in 1993 he presented with psoriasis of the scalp and several other localizations together with Hallopeau's acrodermatitis continua of the ankle, pathognomonic for psoriasic arthritis. Salazosufapyridin was given. The second case was a 50-year-old man who had major pain in both wrists immediately after falling on the palm of his hands in 1992. Bilateral carpal tunnel syndrome developed which did not respond well to surgery. In 1993, he developed inflammatory synovitis and also had psoriasis mainly located at the elbows. Immunological tests were negative. Cortisone and salazosulfapyridin were not particularly effective and the patient later developed arthritis of the hip and ankle joints. Physicians should be aware of physical trauma as a causative factor in psoriasic arthritis due to the potential legal implications. Criteria for imputability are: single major physical trauma, absence of clinical signs prior to the trauma, continuous clinical course, first signs occurring then predominating at the joint exposed to trauma. The pathophysiology of this type of arthritis is not well understood. Deep Koebner's phenomena could be involved. Activation of substance P has also been hypothesized. PMID- 8728888 TI - [Ulcerated necrobiosis lipoidica associated with insulin-dependent diabetes mellitus. Beneficial effect of corticosteroid therapy by oral administration]. AB - OBJECTIVES: Necrobiosis lipoidica is a rare degenerative disease of dermal connective tissue usually observed in young diabetic patients. Several drugs have been suggested to be useful, but none have been shown to be effective in all cases. CASE REPORT: A young patient with insulin-dependent diabetes had a severe invalidating ulcerated necrobiosis lipoidica. Oral corticosteroids led to a satisfactory regression then stabilization with 6 mg/d prednisone. DISCUSSION: Oral corticosteroids would appear to be effective treatment and should be validated by controlled studies. Corticosteroids should only be used in exceptionally severe forms of ulcerated necrobiosis lipoidica after failure of conventional treatments. Diabetes would be an indication under strict metabolic control. PMID- 8728889 TI - [Riedel's thyroiditis and lymphoma. Diagnostic difficulties]. AB - Two cases of Riedel's thyroiditis demonstrated the difficulties encountered in diagnosis. In the first case, subtotal thyroidectomy was performed due to compressive goiter. The intraoperative macroscopic aspect suggested Riedel's thyroiditis. The pathology report also discussed the diagnosis, despite a major lymphoid component, as dense fibrosis had multilated the thyroid parenchyma. Subsequent clinical course led to the diagnosis of lymphoma. The clinical presentation in the second case was classical with rapid increase in size of a hard thyroid gland. Histology was however atypical showing calcifications and liquid cysts within the fibrous thyroid tissue. The initial signs of Riedel's thyroiditis, a very rare disease, usually suggest neoplasia: ligneous compressive goiter with deep adherences. Histologically, there is invasive fibrous thyroiditis with highly suggestive occlusive phlebitis in certain cases. The origin of the disease remains obscure. The differential diagnosis between a sclerosing form and lymphoma is particularly important for therapeutic management. PMID- 8728890 TI - [Aspirin during pregnancy. Indications and modalities of prescription after the publication of the later trials]. AB - Aspirin, an inhibitor of cyclo-oxygenase, is prescribed in a number of conditions related to abnormal production of prostaglandins including gravidic hypertension. Results of the most recent trials demonstrate that in patients with a past history of pre-eclampsia or intra-uterine growth retardation, a pathological Doppler examination of the uterus, a pathological angiotensin test or an antiphospholipid syndrome, prescription of aspirin at the dose of 100 mg/day can prevent recurrence or development of pre-eclampsia or intra-uterine growth retardation. Treatment should begin as soon as possible during pregnancy, certainly before development of clinical manifestations. After history taking and identification of possible contraindications, bleeding time (Ivy method) is recorded before and after prescription and should be lower than 8 minutes. In case bleeding time exceeds 10 minutes 10 to 15 days after initiating aspirin, doses may be reduced to 50 mg per day or even 50 mg every two or three days to reach the target level. Treatment should generally be continued up to 36 weeks gestation. PMID- 8728891 TI - [Pericarditis secondary to anguilluliasis]. PMID- 8728892 TI - [Strychnine poisoning in rural areas. 73 cases]. PMID- 8728893 TI - [Etiological approach in venous thromboembolic disease]. PMID- 8728894 TI - [Pneumococcal cellulitis, secondary septic arthritis]. PMID- 8728895 TI - [Apropos of rheumatic manifestations of celiac disease: a case of Horton disease?]. PMID- 8728896 TI - [Electrocardiographic anomalies during stress-induced hypoglycemia]. PMID- 8728897 TI - [Hormone replacement therapy in menopausal women previously treated for breast cancer]. AB - Is it possible to give a hormonal replacement therapy to a post-menopausal woman with a past history of breast cancer? As estrogens exert a proliferative effect on mammary cells, it remains a controversial issue. Only few studies have been conducted, generally uncontrolled, and involving only a small number of women given short-duration treatment. The endocrinologic arguments are discussed here. Hormone replacement therapy must be replaced in a general discussion on the potential risks for the breast and uterus, and the beneficial effect on coronary heart disease, osteoporosis, and quality of life. The risks and benefits must be weighted for each woman. The circumstances of prescription of hormone replacement therapy, and of the antiestrogen tamoxifen, which has proven a beneficial effect as adjuvant therapy for the menopausal women with a early breast cancer, must be discussed. Some physicians propose a prospective clinical trial to give an answer. Another way could be improvement of our knowledge about the prognostic factors of breast cancer, and better identification of possible "hormonal independance". PMID- 8728898 TI - [Seroprevalence of markers of viral hepatitis A, B and C in hospital personnel at the Clermont-Ferrand University Hospital Center]. AB - OBJECTIVES: Evaluate risk of hepatitis A, B and C infection and anti HBV vaccination policy in hospital personnel. METHODS: A sample of 440 health care workers (7.5% of the personnel at the Clermont-Ferrand University Hospital) representing 74.5% people directly involved in health care and 25.5% other workers were selected at random and stratified by work classification and age. A questionnaire was used to establish personal data on viral hepatitis status and blood samples were drawn for serological tests. RESULTS: Seroprevalence for hepatitis A was 52% with no significant difference between health care and other workers. For hepatitis B, 88.3% of the population had been vaccinated and anti HBs titre was > or = 10 mIU/ml for 91.6% and > or = 50 mIU/ml for 86.1%. Seroprevalence for anti-HBc was 7% and none of the subjects were positive for HBs antigen. Anti-hepatic C antibodies were found in 2 health care workers (0.7%). CONCLUSION: These findings emphasize the need to persue further preventive actions against hepatitis A, B and C and the requirement for continued efforts in elementary hygiene. PMID- 8728899 TI - [Genetic hemoglobin diseases. Prevention at centers for family planning and education of maternal-child protection in Marseille]. AB - OBJECTIVES: Diseases due to inherited hemoglobin disorders represent serious medical, social, and economic problems in the region of Marseille. The only effective treatment for such diseases is allogenic bone marrow transplantation. About 200 patients with either thalassemia, sickle cell or sickle cell-beta thalassemic diseases are regularly seen in local hospitals. All of these patients come from parts of the world where genetic hemoglobin disorders are endemic. METHODS: At this time, the only approach for reducing the number of affected children born is preventive. This depends upon education, the detection of carriers, genetic counselling and sometimes, prenatal diagnosis. We have organised a program of prevention supported by a grant from the DISS (Direction des Interventions Sociales et Sanitaires) in the context of visits made to the PMI (Prevention Maternelle et Infantile). This initiative concerns women presenting for consultations for three reasons: for a prenuptial check-up, for a pregnancy, and for prescription of contraceptives. RESULTS: In each of these three situations a check-up is obligatory and, for natives of countries where hemoglobin disorders are common, a hemoglobin test is recommended. If this test reveals an abnormality, the partner or husband is also tested, if he is willing. Couples who are both carriers are given genetic counselling. CONCLUSION: This preventive initiative has yielded valuable results so we hope to follow-up on the approach and extend it to other centers. Such screening, based upon the geographic origins of patients, can be implemented in the course of a consultation by any doctor. PMID- 8728900 TI - [Non-surgical treatment of perforations of the thoracic esophagus]. AB - OBJECTIVES: Thoracic oesophageal perforations are life-threatening conditions requiring immediate treatment. The type of treatment remains however controversial. The aim of this study was to evaluate the efficacy of the nonoperative management in this disease. METHODS: Six patients were admitted in our surgical unit for thoracic oesophageal perforations during the study period. One perforation was spontaneous (Boerhaave's syndrome). One had complicated an oesophageal ulcer. Two were secondary to caustic lesions. Two were secondary to instrumental dilations. The diagnostic and therapeutic delay was < or = 1 day in 5 cases. In one case the delay was 4 days but the mediastinitis was well tolerated. The treatment was based on antibiotics, nasoesophageal tube suction, and total parenteral nutrition. RESULTS: There was no death in this group of patients. The mediastinitis healed in all cases. In the follow-up period two patients did not have nutritional restriction after healing. Three patients complained of oesophageal stenosis which required oesophageal replacement (n = 2) and dilation (n = 1). One neoplastic stenosis required an endoprosthesis. CONCLUSION: Nonoperative treatment is feasible and safe in selected cases of thoracic oesophageal perforations: perforations diagnosed early and confined to the mediastinum, instrumental perforations, and perforations diagnosed late but well tolerated. PMID- 8728901 TI - [Intermittent chyluria in a young man]. AB - Chyluria is the passage of chylus into urine resulting in fistulization through the lymphatic system and the urinary system. This rare condition is usually caused by filaria infestation or malformations, neoplasia or trauma. We report a case of a 18-year-old man. The patient presented milky urine which had appeared after angiography following minor leg trauma. Physical examination revealed asymmetry of the face and cutaneous dyschromia. Blood tests revealed hypogammaglobulinemia and altered CD4/CD8 ratio (0.6). Urine tests showed proteinuria (30 mg/dl), lipiduria (triglycerides 750 mg/dl) and density of 1025. Renal function was normal. Abdomen computed tomography and urography were normal. Cystoscopy revealed the presence of milky urine in the bladder and selective catheterization revealed that the origin was the right ureter alone. Ascendent pyelography did not reveal any malformation of the urinary tract; but after this the chyluria spontaneously disappeared. The patient was rehospitalized 3 months later for recurrence. Lymphography was then performed and revealed a dilated lymphatic network with minute lacunar images projecting into the right kidney. Chyluria again disappeared spontaneously and recurred sporadically over the next two years in a patient who remained in good physical condition. The etiology of chyluria in a patient without filaria infestation is problematic, particularly when the most common causes (tuberculosis, neoplasia, trauma) are excluded as in our case. The asymmetry of the face, together with cutaneous dyschromia and the presence of a subarachnoidea cyst in the right temporal region suggested our patient had multiple congenital malformations. PMID- 8728902 TI - [Uterine rupture during labor following surgical hysteroscopy]. AB - Two cases of obstetrical uterine rupture after operative hysteroscopy without perforation are described. One was treated for uterus septus, the other one for synechia. However, the uterus could have been fragilized in both cases: one had been perforated by a uterine device, the other case had a past history of repeated curetage. A review of the literature shows 7 other similar cases, published over last ten years, 6 of them having a past history of perforation during the operative hysteroscopy (5 cases) or by an intra-uterine device. These obstetrical complications, although rare, can lead to consider a uterus treated by hysteroscopy as being at risk for obstetrical rupture. PMID- 8728903 TI - [Parvovirus B19 infection. New physiopathological approaches]. AB - Knowledge of the pathophysiological mechanisms governing Parvovirus B19 infection has been modified with the recent discovery of its cellular receptor. The cell lines susceptible to infection can be identified on the basis of the nature and distribution of the virus in the human organism. The Parvovirus B19 can undergo two types of cycles within the host cell. The first is a complete cycle within the infected cell leading to cell death by lysis. In the second, the cycle aborts and cell death is caused by an association of immunological phenomena and the accumulation of viral proteins toxic to the cell. It is not known why complete cycles are restricted to a single erythrocyte line. These new data not only give a more precise explanation of the classical clinical expression of the disease, but also suggest possible mechanisms for the pathogenic power of the Parvovirus B19, some of which have already been proposed on the basis of clinical observations. PMID- 8728904 TI - [Liver transplantation. Current aspects]. AB - The development of liver transplantation has led to technical refinements such as liver graft reduction or splitting for transplantation in two recipients, the use of living related donors, and the temporary orthotopic transplantation of auxiliary liver grafts. Regarding immuno-suppression, new drugs are currently under evaluation, such as tacrolimus (FK 506) which could be effective on some severe rejections. On the other hand, the indications of liver transplantation have been revisited according to clinical results. In France, the increasing development of organ and tissue transplantation has led to the creation of the Etablissement Francais des Greffes, which has the following missions: managing the waiting lists, supporting organ donation, ensuring sanitary safety, and evaluating transplantation activities. Future alternatives to allogenic liver transplantation are mainly represented by xenografting with monkey or pig livers, hepatic assistance and bioartificial liver, and gene therapy. PMID- 8728905 TI - [Cellulitis of the thigh caused by Haemophilus influenzae in an adult]. PMID- 8728906 TI - [Sinus histiocytosis of the Rosai-Dorfman type following Hodgkin disease: efficacy of interferon-alpha]. PMID- 8728907 TI - [Insulin-dependent diabetes mellitus: can body height of patients influence the occurrence of nephropathy?]. PMID- 8728908 TI - [Hemichorea induced by oral contraceptives]. PMID- 8728909 TI - [The Satchmo syndrome]. PMID- 8728910 TI - [Laparoscopic approach in hepatic surgery: segmentectomies II+III]. PMID- 8728911 TI - [Determination of diazepam concentration in maternal and fetal serum after intravenous administration during active phase of labor and its effects on neonates]. AB - OBJECTIVE: To investigate the effects of diazepam on the neonates administered intravenously during the active phase in labour. METHODS: Sixty normal term parturients during the active phase of labour with the cervical dilatation to 3-5 cm were randomly divided into two groups, study group (n = 30) and control group (n = 30). In study group 10mg diazepam was administered intravenously. The concentrations of diazepam and demethyldiazepam in maternal and cord serum were measured by high performance liquid chromatography and neonatal arterial blood gas and Apgar scoring were determined immediately after birth. RESULTS: At delivery, the mean level of diazepam in umbilical cord serum was 947 +/- 314 micrograms/L, markedly higher than that of maternal serum which was 488 +/- 300 micrograms/L, while the mean levels of demethyldiazepam were not significantly different in maternal and fetal serum. There was no marked difference between the two groups in fetal arterial blood gas and acidbase status. However, the rates of respiratory depression and muscle tone inhibition in the neonates of study groups (63.3% and 26.7% respectively) were significantly higher than those in the control group (30% and 3.3% respectively) (P < 0.05, P < 0.01). CONCLUSIONS: Use of diazepam in pregnant women during active phase of labour may cause depression of neonatal respiration. PMID- 8728912 TI - [Time selection of cesarean section in preeclampsia]. AB - OBJECTIVE: To study the relationship between the time of cesarean section and the outcome of mothers and newborns. METHODS: One hundred and three pregnant women suffered from preeclampsia (< 37 weeks 34, > or = 37 weeks 69) and 109 newborns (twins 6) were enrolled for study. The rates of neonatal asphyxia, perinatal mortality and morbidity such as pneumonia, respiratory distress syndrome (RDS) and meconium staining amniotic fluid were observed. RESULTS: The cesarean section rate was 83.06% in preeclampsia cases and it was 94.44% in cases with gestational age less than 37 weeks. There was no maternal death. Neonatal asphyxia rate was 6.67%, and the perinatal mortality rate was 6.67%. After 37 weeks the perinatal mortality rate was 1.41% (P > 0.05) and neonatal asphyxia rate 28.17% (P < 0.01). The rate of pneumonia and meconium staining amniotic fluid were obviously increased. CONCLUSIONS: These observations suggest that fetal lung maturation tended to ahead of gestational age in pregnancy induced hypertension. After 37 weeks complications of both mother and baby increased. In severe preeclampsia cases, pregnancy should be terminated at 34-36 gestational weeks, and cesarean section is the first choice. PMID- 8728913 TI - [Study on the perinatal infection caused by Chlamydia trachomatis]. AB - OBJECTIVE: Study on the frequency and outcome of chlamydia trachomatis (CT) infection of the pregnant women and their babies. METHODS: Two hundred and seventy eight pregnant women and their 79 infants were studied with McCoy cell culture, polymerase chain reaction (PCR) and DNA sequence analysis. Isolation of CT was performed in McCoy cell culture, and iodine-stained glycogen-positive inclusions were considered to be positive. CT DNAs were extracted by modified NaI method. Two sets of primers derived from CT common plasmid and major outer membrane protein genes were used in the PCR. Dideoxy-mediated chain-termination method was used for DNA sequence analysis. RESULTS: 10.8% (30/278) of the cervical cultures were positive for CT, while the positive rate by PCR was 14.0% (39/278). The vertical transmission rate of CT was 55.0% (11/22). The sequences of DNA amplified from a maternal-infant pair were matched perfectly. The incidence of puerperal endometritis in CT positive women was 28.6%, much higher than 8.1% in those CT negative mothers. Neonatal conjunctivitis and pneumonia in group of CT positive mothers were 45.0% and 30.0%, respectively, much higher than 18.3% and 8.3%, respectively, in that of CT negative mothers (P < 0.05). CONCLUSION: Perinatal infection caused by CT is very common in China. PMID- 8728914 TI - [The relationship between maternal body weight index and fetal weight and delivery mode]. AB - OBJECTIVE: To study the relationship between maternal body weight index (BWI) and fetal weight and delivery mode. METHOD: Eight hundred and seventy two normal pregnant women were studied from Jan. 1993 to Dec. 1993 BWI [BWI = weight(kg)/Height(m2)] was calculated in each antenatal visit. The comparing analysis of BWI was done. The study group included 126 mothers with fetal weight > or = 4 000g, the control group consisted of 126 mothers with normal fetal weight. RESULTS: In term pregnancy, the BWI of study groups was significantly higher than that of control group. (P < 0.01). When BWI > or = 27, the rates of macrosomia and cesarean section were markedly increased (P < 0.01). CONCLUSION: Maternal BWI is a valuable index for estimating fetal weight. PMID- 8728915 TI - [The prospective study of trial labor following previous cesarean section]. AB - OBJECTIVE: To investigate the trial of labor following previous cesarean section. METHODS: Three hundred and sixty one cases of pregnant women with history of previous low segment transverse cesarean section were prospectively studied on mode of delivery. 158 cases delivered by elective repeat cesarean section. 203 cases were allowed a trial of labor. RESULTS: 183 out of 203 patients had successful vaginal deliveries. One patient with uterine scar partly ruptured was noted on uterine exploration after delivery. There was no maternal death. The rate of spontaneous vaginal delivery was 79.2%. The rate of vacuum extractor and low forceps deliveries was 20.8%. The 1 minute Apgar score of 170 newborns was > or = 8. Neonatal asphyxia was found in 13 newborns and there was one neonatal death due to congenital abnormality. CONCLUSIONS: Carefully selecting the indication of trial labor and monitoring the labor course are very important for increasing successful vaginal delivery rate and reducing repeat cesarean section. PMID- 8728916 TI - [The quantitative and morphological analysis of placenta and brain in rabbits with intrauterine fetal growth retardation]. AB - OBJECTIVE: To observe the changes of quantitative and morphological structure in placenta and fetal brain of rabbits with intrauterine fetal growth retardation (IUGR) resulted from exposure to cigarette smoking. METHODS: The pure species of 23 pregnant New Zealand rabbits were used and divided into three groups. (1) group A: 9 rabbits were exposed to cigarette smoking everyday in the whole period of pregnancy; (2) group B: 7 rabbits were exposed to cigarette smoking everyday in the late half period of pregnancy; (3) group C: no smoking, as control group. The villus area of placenta, the capillary area in villus and the thickness of granulosa layer in fetal brain were studied by histomorphometric parameter. RESULTS: It was showed that the placental villus area was decreased significantly in group A than that of the control group. The capillary area in villus of both group A and group B was less than that of the group C. The thickness of granulosa layer in cerebrum and cerebellum of group B is less than that of the control group. Nevertheless, only the thickness of granulosa layer in cerebrum of group B decreased significantly than that of the control group. CONCLUSIONS: The development of cerebrum in fetal rabbits were influenced significantly by smoking. The development of cerebellum was influenced by smoking mainly in the early half period of pregnancy. It means that the most important time for cerebellum development is the early period of pregnancy and both the early and late half period of pregnancy were important for the cerebrum development. PMID- 8728917 TI - [Responses of somatostatin, beta-endorphin and dynorphin A to A glucose load in two groups of women with polycystic ovarian syndrome]. AB - OBJECTIVES: To investigate the relationship between elevated luteinizing hormone (LH), hyperinsulinemia and neuropeptides in patients with polycystic ovarian syndrome (PCOS). METHODS: An oral glucose (100g) tolerance test (OGTT) was performed in 15 normally menstruating women (control) and 30 PCOS women with LH/follicle stimulating hormone (FSH) ratio greater than 3 (group 1) and 25 PCOS subjects with the ratio < 3 (group 2). The responses of insulin, somatostatin (SS), beta-endorphin (beta-EP) and dynorphin A (Dyn A) during OGTT were measured by RIA. RESULTS: In basal state, significant negative correlations were found between LH and SS (r = -0.51, P < 0.05) in group 1 and between LH and beta-EP (r = -0.49, P < 0.05) in group 2. During OGTT, PCOS women had a greater beta-EP and Dyn A responses in group 1 and an impaired SS response in group 2 as compared with the control. CONCLUSION: These data suggested lower endogenous SS, higher beta-EP and Dyn A may lead to elevation of LH and insulin secretions in patients with PCOS. PMID- 8728918 TI - [Diagnosis and treatment of postmenopausal uterine bleeding by hysteroscopy]. AB - OBJECTIVES: To evaluate the value of hysteroscopy in diagnosis and treatment of patients with postmenopausal uterine bleeding. METHODS: One hundred and thirty five patients with postmenopausal bleeding hospitalized in our department from July 1990 to July 1994 were examined by hysteroscopy. The results of 124 cases were compared with pathological findings obtained during uterine curettage. Thirty-nine patients were operated simultaneously under hysteroscopy. RESULTS: Under hysteroscopy, atrophic endometrium, endometrial polyps, cervical polyps, endometrial carcinoma, endometrial hyperplasia, submucous myoma were diagnosed in 62 cases (45.9%), 25 cases (18.5%), 13 cases (9.6%), 13 cases (9.6%), 11 cases (8.2%) and 5 cases (3.7%) respectively. The remaining 6 (4.5%) had IUD insertion. These findings were confirmed pathologically in 115 out of 124 cases (92.7%). There were 9 cases misdiagnosed by hysteroscopy. 6 cases had false negative results during previous curettage. Twenty-two cases of endometrial polyps, 13 cases of cervical polyps and 4 cases of pediculated submucous myoma were removed hysteroscopically without any complication. CONCLUSIONS: Hysteroscopy is an effective method for identifying the causes of postmenopausal uterine bleeding, which is superior to curettage of uterus. For some selected patients hysteroscopic operations can be performed at the same time. PMID- 8728919 TI - [Effects of gestrinone on experimental endometriosis in rabbits]. AB - OBJECTIVE: To study the therapeutic effects of gestrinone on experimental endometriosis in rabbits. METHODS: Mature female rabbits were laparotomized under sterile condition and three or four segments of endometrium (7 mm x 7 mm) were sutured to the mesentery of uterus. Seven weeks after operation these animals were divided into 5 groups: control; treated with danazol 20mg/kg; gestrinone 0.06, 0.12, 0.24 mg/kg for 4 weeks respectively. At the end treatment they were laparotomized again. The ectopic endometrial implants were weighted and their morphologic features were analysed quantitatively by semi-automatic image analysis system, meanwhile serum follicular stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E2) and progesterone (P) concentrations were determined by radioimmunoassay. RESULTS: Decreased weight and atrophy of ectopic endometrial implants were shown in all experimental groups, except for control and 0.06 mg/kg gestrinone treated-group. In 0.12, 0.24 mg/kg gestrinone treated group, except for P serum FSH, LH and E2, concentrations decreased significantly. The degree of inhibition on ectopic lesions of 0.12 mg gestrinone group was similar to that of danazol group. CONCLUSIONS: Gestrinone inhibits the growth of ectopic endometrial implants in a dose-dependent manner and showed a significant therapeutic effects on experimental endometriosis in rabbits. PMID- 8728920 TI - [A quantitative study on nuclear DNA content and morphological parameters in ovarian cystadenocarcinoma]. AB - OBJECTIVE: To study nuclear DNA content and morphological parameters in ovarian cystadenocarcinoma. METHODS: Nuclear DNA content and morphological parameters were quantitatively determined by image analysis technology in 56 cases of ovarian cystadenocarcinoma, 12 benign ovarian cystadenoma and 10 normal ovarian epithelial tissue. RESULTS: DNA content of ovarian cystadenocarcinoma was the highest among the three groups; and there were significant differences between the groups (P < 0.01). In the aspect of morphometry, nuclear circle was greatly different among the three groups (P < 0.01). There was no significant variances in nuclear parameter and area (P > 0.05). Nuclear DNA content of ovarian cystadenocarcinoma was related to its histopathological classification, histological grade and clinical stages (P < 0.01, P < 0.05). There was a reverse relationship between the DNA ploidy level and clinical prognosis. The 2-year survival rate of ovarian cystadenocarcinoma with diploidy/near diploidy pattern was higher than that with aneuploid pattern (P < 0.01). CONCLUSIONS: Nuclear DNA content of ovarian cystadenocarcinoma is a useful marker in reflecting the biological behavior and is of practical value in diagnosis, the choice of treatment methods and in the evaluation of results and prognosis. PMID- 8728921 TI - [Late bladder complications following radiotherapy of carcinoma of the uterine cervix]. AB - OBJECTIVE: To analyse the causes, clinical features, treatment and prevention of late bladder complications following radiotherapy of carcinoma of the uterine cervix. METHOD: This clinical retrospective study included 378 cases of irradiation cystitis following radiotherapy for carcinoma of the uterine cervix from February, 1958 to October, 1987. RESULTS: The incidence of irradiation cystitis was 3.2% and was not related to age, FIGO staging and the methods of intracavitary irradiation (conventional or afterloading). It was related to the external appearance of the local lesion; 66.5% of the tumors of patients with late bladder complications was of the exophytic type prior to radiotherapy. 51.1% of the patients developed late bladder complications in a period of 2-5 years after radiotherapy; 16.7% in 5-10 years and only 3.4% within the first years. The patients with irradiation cystitis had received a 10% higher dosage than that routine delivered, as found either in intracavitary irradiation of 39.4% of patients or in external irradiation of 31.6% of patients. Based on the clinical features and cystoscopic findings, late bladder complications can be divided into three types: (1) sudden and temporary hematuria (35.7%); (2) persistent hematuria (63.2%); and (3) vesico-vaginal fistula (1.1%). The 5-year survival rate of patients with irradiation cystitis was 90.4% which was higher than the survival rates of all patients treated by conventional intracavitary irradiation (65.7%) and all patients treated by afterloading intracavitary irradiation (70.4%). 3.7% of patients died of late bladder complications. CONCLUSIONS: To decrease the occurrence of late bladder complications and to obtain long term survival: (1) Dosage to the bladder should be adequate, and over dose to the bladder should by all means be avoided; (2) Patients are to be taught to observe indicated personal care during radiotherapy as well as in the follow-up period; (3) Early institution of proper conservative treatment of cystitis is beneficial. PMID- 8728922 TI - [Advances in the study on adenoma maligum of the uterine cervix]. PMID- 8728923 TI - [Clinical significance of ovarian cancer related oncogenes and tumor suppressor genes]. PMID- 8728924 TI - [Surgical treatment of craniosynostosis]. AB - Craniosynostosis is the term that designates premature fusion of one or more cranial sutures that causes severe cranial deformities. The clinical features and the operative technique were introduced and the etiology and the key points for a successful operation were discussed in this paper. PMID- 8728925 TI - [Anatomical study and clinical application of extended deltopectoral flap]. AB - The study on the anatomy of the deltopectoral flap that extended to the area of the neck, should or back was carried out in 30 fresh adult cadavers by means of dissecting under microscope, MO-Pd radiography, carbon-ink injection of arteries. We observed the origin of cutaneous arteries of the flap and the running direction, length and diameter of vessel branches and its anastomotic network. The clinical applications and the prospect of the flap are described. PMID- 8728926 TI - [Clinical use of the C-shaped cross finger flap]. AB - The C-shaped cross finger flap is an axial flap based on the digital vascular bundle either distally or proximally. We have successfully used this flap to repair different defects of the finger in 12 cases. The flap can be made rather big and the operation is easy. The surgical technique and the advantages and disadvantages are described. PMID- 8728927 TI - [The applications of machinable bioactive glass ceramics in maxillofacial augmentation]. AB - Machinable bioactive glass ceramics (MBGC) has been employed in maxillofacial augmentation as a substitute for bone grafts in 36 patients with satisfactory results. Two years' follow-up did not show inflammatory reaction and rejection of the implant. Clinical applications of MBGC proved its reliability. PMID- 8728928 TI - [The effect of different topical agents on pseudomonas infection of burn wound]. AB - 48 Wistar rats were scalded resulting in 15% TBSA full-thickness burn. Pseudomonas aeruginosa (10(9)/ml was inoculated to the wounds. The rats were divided into 6 groups. 5 different topical agents were separately applied to the wounds constituting 5 groups, and no drug was used in control group. Through observing the appearance of wound, the quantity of bacteria in the subeschar tissue and histo-pathological changes, we confirmed that AgSD-ZnSD-A cream was a satisfactory topical agent. PMID- 8728929 TI - [Ultrastructural changes in pulmonary microvascular damage in rats inflicted with burn, blast and combined burn-blast injury]. AB - The pathological changes in pulmonary microvasculature in rats inflicted with overpressure blast, 15% third degree burns and combined burn-blast injury, respectively, were studied with electron microscopy and ultrastructural morphometry. Results indicated that the pathological changes in pulmonary microvasculature in 15% third degree burns were slight, and mainly restricted to ultrastructural level. While in severe blast and burn-blast injuries, the damage of pulmonary microvasculature including endothelial swelling and regeneration and dispersal of arteriole wall structure, was prominent. These changes form the basis of pulmonary hemorrhage and edema. Apart from vessel structural damages, arteriole constriction and capillary closure were also observed during 2 to 4 h postinjury. The vessel damage and dysfunction were more severe in combined injury than in blast injury alone. The mechanisms needed to be studied further. PMID- 8728930 TI - [Effects of resuscitation with hypertonic sodium lactate dextran 70 on cardiac function in severely burned dogs]. AB - This comparative study was performed to evaluate the effects of resuscitation of burn shock with hypertonic sodium lactate dextran 70(HLD solution: Na+250 mmol/L and 6% dextran 70) and lactate Ringer' solution on cardiac function, lipid peroxide and SOD activity of heart tissue. Dogs with 35% TBSA third degree burns received, one hour postburn, either HLD resuscitation (HLD group, n = 6) in the amount of 19.6 ml.kg-1/g hours, followed by LR 6 ml.kg-1/% TBSA, or LR resuscitation (LR group, n = 6) in the amount of 8 ml.kg-1/% TBSA LR solution. The results of this experiment showed that cardiac index (CI) of HLD group was much higher than that of LR group at 4, 8, 12, 24 hours postburn. The dp/dt max and-dp/dt max of HLD group were much higher than that of LR group at 12.24 hours postburn. Level of MDA in heart tissue of HLD group (1.74 +/- 0.28 10 mol/g heart tissue) was much lower than of LR group (3.23 +/- 0.56 10 mol/g heart tissue) at 24 hours postburn (P < 0.01). Meanwhile, SOD activity in heart tissue of HLD group (157.49 +/- 32.23 u/mg heart tissue) was much higher than that of LR group (46.88 +/- 16.28 u/mg heart tissue) (P < 0.01). These results suggested that HLD resuscitation might be beneficial in improving postburn cardiac function and attenuating postburn oxidant-induced lipid peroxidation in heart tissue not only by its "oxygen scavenger" effect, but also by its elevation of SOD activity of heart tissue. PMID- 8728931 TI - [Large-scale culture of human keratinocytes]. AB - We propose a modification of the conventional keratinocyte subculture method, showing a significant improvement in the colony growth capacity of subcultured keratinocytes. This method utilized mouse 3T3 fibroblasts and MCDB153 medium, and it has been shown that they could improve the colony growth of human keratinocytes. Compare with the conventional procedure, this culture method was shorter in time, and resulted in a higher colony growth capacity of cells, reaching confluence 3 days earlier. In the treatment of extensively burned patients using cultured epidermal sheets, the time required for their production remains the main problem. Thus, the significantly reduced time to obtain confluent keratinocyte sheets with our method is very important for the treatment of large burn wounds. PMID- 8728932 TI - [Viability and antigenic changes in the skin after preservation under different temperatures]. AB - Oxygen consumption and succinate dehydrogenase contents were determined to compare mean viabilities (%) of human cadaver and guinea pig skin stored under different temperature: 4 C, -20 C, -80 C, and -196 C. The results showed that the mean viability (%) of skin cryopreserved under -196 C was superior to those under 4 C, -20 C, and -80 C (70.8%, 61% respectively for -196 C, P < 0.05 or 0.01). Microimmunoelectrophoresis and rocket immunoelectrophoresis were performed for related antigens and antibodies. The results of microimmunoelectrophoresis showed that the antigenicities were different for four groups. Antigenicity of fresh skin was highest. The higher the storage temperature, the lower the antigenicity of skin was. PMID- 8728933 TI - [Study on the retrofascial space of the dorsum nasi and its clinical significance]. AB - Fifty-seven adult and two children cadavers were dissected to study the position, size and structure of the retrofascial space of the dorsum nasi. The space lay between the periosteum, perichondrium and the fascia dorsum nasi. The shape of the space is like a pyramid. The space averaged 3.09 +/- 0.66 cm in length. The anatomical characteristics, individual variation and clinical significance of this space were discussed. PMID- 8728934 TI - [Anatomic structure of the soft tissue of nasal dorsum and its applications in augmentation rhinoplasty]. AB - The nasalis--procerus aponeurosis and its clinical applications in augmentation rhinoplasty were described in this paper. According to our experience, the silicone frame can be put in three anatomical layers of the nasal dorsum for augmentation. The paper showed the manipulating points and problems of the operations done through various anatomical layer, and also pointed out that good results can be obtained with the silicone frame being put on or beneath the nasalis--procerus aponeurosis. PMID- 8728935 TI - [Rising and trends of endoscopic cosmetic plastic surgery]. PMID- 8728936 TI - Transseptal methods for percutaneous balloon valvoplasty simultaneously with radiofrequency catheter ablation. AB - Percutaneous balloon mitral valvoplasty (PBMV) and radiofrequency catheter ablation (RFCA) have been used in the treatment of mitral stenosis (MS) and supraventricular tachycardia. The techniques of PBMV and RFCA yield better results with the development of interventional cardiology, but there is no report about PBMV performed simultaneously with RFCA in the same patient. Seven patients with mitral stenosis and Wollf-Parkinson-White (W-P-W) Syndrome were successfully treated with PBMV and RFCA by transseptal methods. LA, LAP, mPG and mPA were decreased from 43.4 +/- 4.6mm, 21.8 +/- 6.8mmHg, 21 +/- 7.7mmHg and 45.7 +/- 16.5mmHg to 39.2 +/- 3.7mm (P < 0.05), 12.7 +/- 4.5mmHg, 12 +/- 3.7mmHg and 32.3 +/- 9mmHg (P < 0.01). MVA was increased from 0.96 +/- 0.33cm2 to 1.7 +/- 0.80cm2 (P < 0.01). delta wave disappeared in 12-lead surface EKG and SVT could not be induced in electrophysiological study after the treatment. The overall time of the procedure for this series was 93 +/- 34 minutes and fluoroscopy time was 23 +/- 7 minutes on the average. Radiofrequency energy applications were 3 +/- 2 times each procedure. PBMV and RFCA are safe and highly effective in the treatment of MS and W-P-W syndrome. The results in the present study proved the feasibility of the combined use of PBMV and RFCA. We prefer a first choice of PBMV and then RFCA in order to avoid aggravation of the hemodynamics due to mitral stenosis. The results also showed that of overall procedure and fluoroscopy only took a short time for this series. We suggest that it could be used as a routine method for the treatment of mitral stenosis complicated by W-P W syndrome. PMID- 8728937 TI - Long-term survival of heterotopic heart transplants in allogenic rats without using anti-rejection drugs after inducing immune tolerance. AB - Graft rejection is a baffling problem in organ transplantation which has so far no successful resolution. Present antirejection therapy is expensive, toxic, susceptible to infection, oncogenesis and atherosclerosis. With the use of 2 extremely histoincompatible strains of rats as model, it was found that when six donor splenocytes preconditioned by an extract from the culture medium of streptomyces caespitosus were injected into the host, after 2 weeks immune tolerance heterotopic transplanted donor hearts could survive normally more than 100 days without using any anti-rejection drug. Their mean survival time was 141.4 +/- 2.56 days, while that in the control group was only 11.7 +/- 3.48 days (P < 0.005). Thus, the present research offers some bright prospects in the area of organ transplantation. PMID- 8728938 TI - A study of mRNA expression of glucose transporter gene in rats with diabetes mellitus. AB - The study was proceeded on the mRNA expression of the GLUT-1 and GLUT-4 gene in the alloxan-induced diabetic rats by the method of dot-blot hybridization analysis. The results showed that the levels of the GLUT-4 mRNA expression were reduced in the muscle tissue of the diabetic rats while its levels in the same tissue of insulin treated diabetic rats increased. It was suggested that the reduced GLUT-4 mRNA expression which could affect its posttranslational products may be one of the causes of the insulin resistance. PMID- 8728939 TI - An animal study on transmission of hepatitis B virus through mosquitoes. AB - Three gena of mosquitoes were fed with human blood infected with hepatitis B virus (HBV) artificially. When blood in the stomach was completely digested, mosquitoes were used to bite monkeys Tupaia Belangeri. A month after biting, venous blood samples were taken from the monkeys to detect serum HBsAg, HBeAg, Anti-HBs and Anti-HBc. Among 29 test monkeys, 9 were HBV infective serum markers positive. In 4 of 16 test monkey liver biopsies, acute inflammatory changes were found with Hematoxylin-Eosine staining. In the hepatic cells, HBsAg and HBcAg were detected by immunohistochemical double-labeling assays, and HBV-DNA was also positive by in situ hybridization in monkey liver sections. These results provided an evidence for the possibility of transmission of HBV through mosquitoes and suggest its epidemiological significance in mosquito infested areas. PMID- 8728940 TI - Ultrastructure of parietal cells before and after extended parietal cell vagotomy in patients with duodenal ulcer and their complications. AB - Since 1979 we have treated 149 patients with duodenal ulcer by extended parietal cell vagotomy (EPCV). Electron microscopy of parietal cells from gastric body mucosa and gastric acid secretory test were carried out in 22 randomly selected patients before and after EPCV. It was shown that before EPCV the ultrastructure of parietal cells presented the feature of vigorous section identical to the hyperfunctional secretion of gastric acid. The ultrastructure of parietal cells presented the feature of acute depression of secretory function, which was identical to a significant reduction of secretory function of gastric acid 0.5-6 months after EPCV. Two to 10 years after operation, however, especially over 10 years after EPCV, the ultrastructural changes of parietal cells gradually returned to normal, but gastric acid secretion remained at a low level, without preoperative secretory capacities. The results of this study provide a histological basis for the application of extended parietal cell vagotomy in the treatment of duodenal ulcer and its complications. PMID- 8728941 TI - Epidemiology of anti-human T-cell leukemia virus type I antibody and characteristics of adult T-cell leukemia in China. AB - We analyzed the data from seroepidemiologic survey of anti-HTLV-I serum antibody (HTLV-1 Ab) and 16 adult cases of T-cell leukemia (ATL) in China (1984-1994). The results showed that the positive rate of HTLV-I Ab of the population of China was 0.54%, the birth or residence places of HTLV-I carriers and all the patients with ATL were mainly distributed in the coastal provinces and the northeast China, and a small endemic area of HTLV-I and ATl was found in Fujian Province (HTLV-I Ab positive rate, 2.1%). The HTLV-I in China may be transmitted from Japan or originated from China. The clinical characteristics of the cases included onset in adulthood; frequent skin lesions, lymphadenopathy and hepatosplenomegaly; no mediastinal mass; the ATL cells from peripheral blood with polymorphism nuclei and mature T cell immunophenotype HTLV-IAb positive in most of the cases tested (5/8); mostly acute type; with a median survival period of 3.5 months. PMID- 8728942 TI - Findings of cholangiopancreatography in pseudotumorous pancreatitis. AB - The cholangiopancreatographic findings in 22 patients with pseudotumorous pancreatitis (PTP) were evaluated on the basis of endoscopic retrograde cholangiopancreatography (ERCP) and percutaneous transhepatic cholangiography (PTC). All patients were found with a focal mass in the pancreas based on the findings of ultrasound and/or CT. However, only one patient had been diagnosed correctly as suffering from chronic pancreatitis by ERCP. Normal pancreatic duct was found in 4 patients. There were five patients having abnormal pancreatic ducts, but findings of ERCP could not provide definite diagnosis. In the remaining 12 patients the abnormalities of pancreatic and/or bile ducts were strikingly similar to those found in pancreatic carcinoma. Double duct sign was also seen in one patient. All these 12 patients were incorrectly diagnosed as having pancreatic carcinoma. It is concluded that ERCP/PTC has little value in detecting PTP. PMID- 8728943 TI - Prospective studies on the relationship between the 50 g glucose challenge test and pregnant outcome. AB - The 50 g oral glucose challenge test (50gGCT) was performed on 622 pregnant women, and 75 g oral glucose tolerance test (75gGTT) was further done on subjects with screening tests value of > or = 7.78 mmol/L. The results showed that there were 16.56% (103/622) women with screening value of > or = 7.78 mmol/L, among whom, 32 were identified as having gestational impaired glucose tolerance (GIGT) and 12, gestational diabetes mellitus (GDM) by confirmatory test of 75gGTT. The sensitivity of 50gGCT was 42.72%(44/103). The incidences of edema-proteinuria hypertension syndrome (EPH-syndrome), premature rupture of membranes, fetal macrosomia, operative deliveries and perinatal morbidity were higher in women with GIGT/GDM than in women without GIGT/GDM. It suggests that 50gGCT is an ideal method of screening for GDM and should be performed on all pregnant women. PMID- 8728944 TI - Induction of osteogenesis by bovine platelet transforming growth factor-beta (TGF beta) in adult mouse femur. AB - TGF-beta has been shown to be one of the most important regulators in chondrogenesis and osteogenesis. Daily subperiosteal injection of TGF-beta, which was extracted and purified from bovine platelets, into adult mouse femur resulted in proliferation and differentiation of the mesenchymal cells, chondrogenesis, and endochondral osteogenesis. After cessation of the injection, the endochondral ossification occurred widely, resulting in replacement of the cartilage with new bone. These phenomena demonstrate that exogenous TGF-beta stimulates mesenchymal proliferation and differentiation that occur in early fracture healing and suggest the possibility of clinical application of TGF-beta for fracture repair and bone transplantation. PMID- 8728945 TI - Effect of angiotensin converting enzyme inhibitors on tolerance of nitroglycerin. AB - The aim of this study was to assess whether the angiotensin-converting enzyme inhibitors captopril and enalaprilat could change the tolerance of nitroglycerin in isolated rat aortae. In aortic rings precontracted with potassium chloride, captopril (1 microM) but not enalaprilat (0.1 microM) incubation potentiated the responses to nitroglycerin. It is suggested that captopril can reduce the tolerance of nitroglycerin because it is a sulfhydryl group donor. PMID- 8728946 TI - Study on motor functions of antrum after highly selective vagotomy (HSV) plus mucosal antrectomy. AB - To elucidate the effects of highly selective vagotomy plus mucosal antrectomy (HSV-MA) antral functions, eighteen dogs were randomly divided into control, HSV and HSV-MA group for determining gastric emptying, antral myoelectric activity and antral pressure. The tests were performed 4 months after operation in the HSV MA and HSV group. By roentgenoscopy, the gastric and duodenal shape were normal, and gastric peristalsis was clearly seen in all the dogs in the three groups. The beginning and ending time of gastric emptying in the HSV-MA group were 5.0 +/- 0.06 minutes and 4.0 +/- 0.4 hours respectively, which were similar to those in the control and HSV group (P > 0.05). The frequency of antral myoelectric action potential in the HSV-MA group (3.11 +/- 0.65 cycles/minutes) was significantly lower than those in the control and HSV group (P < 0.05). Intramuscular injection of pentagastrin significantly increased the frequency of antral myoelectric action potential (P < 0.01) in all groups, and eliminated the difference among them before the injection. The antral pressure wave occurred immediately following the antral myoelectric action potential waves. The antral pressure values in the control and HSV-MA group were similar (P > 0.05), and were significantly lower than that in the HSV group (P < 0.05). Intramuscular injection of pentagastrin significantly increased the antral pressure (P < 0.001) in all groups and eliminated the difference among them before the injection. These evidences suggest that the motor function of the reconstructed antrum following HSV-MA nearly reached normal level despite a lower antral myoelectric frequency. Combined with other reports, HSV-MA has a good clinical prospect as an operation for the treatment of chronic duodenal ulcer. PMID- 8728947 TI - Evaluation of hepatocyte growth-promoting factors in treating 1687 cases of fulminant hepatitis. PMID- 8728948 TI - Thrombolytic therapy for acute cerebral vascular occlusion. PMID- 8728950 TI - Ovarian sclerosing stromal tumor with minor sex cord elements. PMID- 8728949 TI - The source of endogenous digitalis-like substance in normal pregnancy. PMID- 8728951 TI - A case of aneurysmal bone cyst of maxillary sinus. PMID- 8728952 TI - [Evaluation of acute flaccid paralysis (AFP) surveillance system in 1994]. AB - Increased by 40.2% compared with 1993, 3142 acute flaccidparalysis cases were reported from AFP surveillance system in 1994. A total number of 1297 counties had AFP cases reported, accounting for 45.1% of total amount of counties. Out of 3142 AFP cases, 307 were confirmed, coming from 225 counties. For children under 15 years old, the reported AFP incidence was 1.1/10(5), non-polio AFP rate was 0.83/10(5), much higher than 0.37/10(5), in 1993. 48.5% of the total AFP cases had double stool specimen taken within 0-14 days after paralysis. 79% AFP cases had been investigated within 48 hours after being reported. Some of the major monitoring indicators of AFP surveillance system were close to the WHO criterion, but the percentage of AFP cases reported within 48 hours of careseeking and AFP cases with follow-up visit after 60-70 days of paralysis were very low. Furthermore, 77.9% of the confirmed cases were diagnosea clinically by residual paralysis, so did the 46.4% of discarded cases. It is in urgent need to increase the percentage of cases classified by laboratory test. PMID- 8728953 TI - [Epidemiological analysis of poliomyelitis cases in 1994, China]. AB - In 1994, the number of reported polio cases was 307 in China. This number was 53% lower than that in 1993, and was the lowest record ever in China. In 1994, the distribution of polio cases was highly dispersed and the number of outbreak was reduced obviously. Of 225 counties which reported polio cases in 1994, polio outbreak occured in 2 counties. Cases due to polio outbreak accounted for 4.2% of the total cases. The distribution of polio wild virus was narrow but the regional difference still existed. In the southeast, high risk region for polio consisting Guangdong, Hainan, Fujian three provinces, the number of polio cases decreased remarkably in 1994. On the contrary, in the southwest region which, consisting Guangxi, Yunnan, Guizhou, Sichuan four provinces, the number of polio cases increased obviously, accounted for approximately a half of the total number in 1994. During the coming 1 to 2 years, we should emphasis on these regions and provinces where high incidence was noticed in our polio eradication program. Those provinces should include Xinjiang, Fujian. Hubei three provinces that polio wild virus was identified in 1994, and Guangxi, Sichuan, Guizhou, Yunnan in the south west region as well. We should further increase Polio vaccination coverage of children especially floating and unplanned ones through strengthening routine immunization and carring out new national immunization day activity. PMID- 8728954 TI - [Evaluation of second National Immunization Day activities, in China]. AB - The second National Immunization Day (December 1994/January 1995) for 0-47 months old children, seemed to be well conducted, for not only the coverage of OPV being increased, but also the continued accumulation of zero-dose children being under controlled. In most of the provinces, it appears that the coverage of the second NID was significantly greater than the first NID. Based on comparison of the number of children immunized by birth cohort, there were 7.9% (1st round) and 4.3% (2nd round) more 1-3 year old children immunized in the 2nd NID than the 1st NID. Most of children missed in the 1st NID were distributed in one year old cohort (they were less than 1 year old the year before), so it is most likely that children less than 1 year old are more at risk to be missed by NID. After the 1st NID, the numbers of zero-dose children were reduced to 59.3%, and the percentage for those zero-dose children less than 1 year old increased from 66% to 84%, so children less than 1 year old should have the high priority to be immunized for achieving expected goal. The second round of second NID was not implemented so well as the first round in some of the provinces. Comparing with the first round, the number of children immunized at second round was decreased in 468 (18%) counties. In addition, the number of expected children to be immunized was less than the registered children, in 37% of the counties; and still there were 19% of the counties in which the number of children immunized in the second NID were less than the children with routine immunization cards. The above mentioned problems need to be solved effectively. PMID- 8728955 TI - [Epidemiological analysis on morbidity of acute flaccid paralysis (AFP) among children under 15 years old in 14 provinces of China]. AB - Five investigations through analysis of hospital records on morbidity of Acute Flaccid Paralysis (AFP) among children under 15 years old in 14 provinces of China have currently been conducted. In this paper, data from a series of studies published in journals or reported to meetings was combined and analyzed based on Meta-Analysis. Comparisons have been made between AFP, poliomyelitis, Guillian Barre Syndrome (GBS) and non-polio AFP morbidities and their distributions by age, sex and month. The morbidity rates for these four categories were 1.41, 0.54, 0.55 and 1.05 per 10(5) respectively. An important conclusion drawn from of this study was that the criterion put forward by WHO, reported rate of non-polio AFP should reach over 1.0 per 10(5) among children under 15, can also be used as an indicator for sensitivity evaluation and monitoring of AFP surveillance system in China. PMID- 8728956 TI - [Poliomyelitis surveillance in Shandong Province from 1991-1994]. AB - After a series of poliomyelitis outbreaks came about in Shandong province from 1988 to 1990, comprehensive measures including establishment of acute flaccid paralysis (AFP) surveillance system, introduction of mass immunization campaign of oral poliovirus vaccine (OPV) and enhancing management of poliomyelitis eradication activity, were carried out. The incidence of poliomyelitis decreased with the reported number of cases 95, 25, 22 and 17, in 1991, 1992, 1993 and 1994, respectively. A geographically sporadic distribution was noticed with no peak on seasonality. No confirmed case caused by wild poliovirus was identified from 1991 to 1994. Reporting incidence of AFP cases among children under 15 years old was increasing gradually with 1.00 per 10(5) population in 1993 and 1.89 in 1994. The results showed that the poliomyelitis eradication activity in Shandong province had reached a final stage but much efforts still need to be made to reach the target on poliomyelitis eradication. PMID- 8728957 TI - [Analysis on poliomyelitis cases in Inner Mongolia in 1990-1993]. AB - Out of 97 AFP cases, 68 were confirmed polio in Inner Mongolia in 1990-1993, in which 44.12% were diagnosed according to laboratory results. In this period, 13 strains of polio virus were isolated and identified from the stool samples of 62 AFP cases. The etiological results suggested that there was no dominant strain of polio virus in the whole region. Serological results showed that the positive rates of IgM antibody against polio virus were 75%, 26% and 0% in 1990, 1991, 1993 respectively which seemed to have a declining trend. PMID- 8728958 TI - [Analysis on the health status of residents from Diseases Surveillance Points in Gansu Province]. AB - We have surveyed a population size of 6633315 from Diseases Surveillance Points (DSP) system in Gansu province for the last eleven years. The annual birth rate was 18.20% with an annual standard mortality rate 545.80/10(5). The annual standard mortality for male and female were 607.53/10(5) and 483.29/10(5) respectively. The major causes of death were Respiratory system diseases, Cardiovascular diseases, Neoplasms, Injuries, Digestive system diseases, Pediatric diseases, Infectious diseases in sequence. In eleven years, there seemed to be a rising trend in the mortalities of following diseases as: Cerebrovascular diseases, Ischemic heart diseases, Rheumatic fever and heart disease, Lung Cancer, Liver Cancer, Cancer of the Esophagus, Intestinal cancer, Cervical cancer, Injury, Congenital abnomalities, to different degrees. However, an obvious descending trend on the morbidity and mortality of infectious diseases was moticed. The average life expectancy was 71.05 years in DSP, with male 69.57 years, and female 72.72 years. Diseases with higher PYLL were Injuries, Neoplasms, Respiratory system diseases and the like. Data suggested not only the prevention andcontrol of infectious diseases, but also the surveillance of injuries and the prevention and control of chronic diseases should be strengthened. PMID- 8728959 TI - [A seroepidemiological study on hepatitis D virus (HDV) infection in Henan Province, China]. AB - In order to investigate the distribution of HDV infection in Henan province, a seroepidemiological study on HDV infection was conducted. A total number of 1182 HBsAg positive serum samples was collected from 13 prefectures and cities between 1991 and 1993. HDV markers (HDAg, anti-HDV, anti-HDV-IgM) were tested with Enzyme Linked Immunosorbant Assay (ELISA). The prevalence rates of HDAg, anti-HDV, anti HDV-IgM and HDV infection rate were 3.0%, 3.5%, 3.5% and 8.1%, respectively. There were no statistically significant differences in the prevalence rates of HDV infection in patients of different sexes but significant differences in the prevalence rates of HDV infection in patients of different ages, especially in age group above 60, were noticed. We also noticed that the infection rates of HDV were 8.7%, 4.6%, 7.7%, 6.3%, 11.9% and 0.0% in the groups of HBsAg carriers, acute hepatitis, chronic active hepatitis, chronic persistent hepatitis, liver cirrhosis and severe hepatitis, respectively. These findings indicated that HDV infection was important in the pathogenesis of chronic hepatitis B and in the exacerbation from liver disease to cirrhosis. PMID- 8728960 TI - [The sero-epidemiological investigation of leptospirosis in Hainan Province]. AB - This paper reported that 529 serum samples were collected from healthy people in ten townships of nine cities or counties of Hainan Province in 1993 and anti leptospira antibody was examined. Results showed that 274 cases were positive (51.8%), indicating the natural infection rates were from 25.9% to 72.2% in different areas. The infection was irrelevant to sex, but was interrelated to age and occupations. There were 15 serogroups of anti-leptospira antibody identified in this study. Bataviae, Grippotyphosa and Icterohaemorrhagiae serogroups were the dominant serogroups, different from that the Grippotyphosa and Autumnalis serogroups as the superior serogroups in the 1970s. Data shows that the main epidemic serogroups are likely to change, which suggests that the surveillance of leptospirosis needs to be strengthened. PMID- 8728961 TI - [Virulent factors of Legionella pneumophila and their uses in molecular epidemiology]. PMID- 8728962 TI - [Clinical and radiographic analysis of 56 cases of mural ameloblastoma]. AB - Fifty-six cases of mural ameloblastoma were ananlyzed clinically and radiographically. They were transformed by ameloblastomous changes from keratocyst, occuring in 28.2% of ameloblastoma. The disease was often seen in patients of 10-29 years old with a mean of 21.8. Most of them were seen in mandible (53/56), especially in ramus (40/50). The radiographic appearance is divided into three subtypes: (1) ramus-mandibular body pattern, including dentigerous (31/56) and non-dentigerous (9/56); (2) sub-apical pattern (12/56); (3) inter-radicular pattern (4/56). The mainpoint of radiographic diagnosis is the appearance of unilocular or multilocular radiolucency of similar size, located in ramus-mandibular body or mandibular body region. The large lesions have a tendency to expand along the mandible. The roots of teeth are absorbed apparently, demonstrating serrated or amputated appearance. PMID- 8728963 TI - [The correlations of two methods in the evaluation of velopharyngeal competence]. PMID- 8728964 TI - [The effect of occlusal force on the formation of experimental cervical wedge shaped defects on human teeth]. AB - Nine pairs of premolar from 7 patients were divided into three groups according to different occlusal forces. Every tooth was tested with pH5 or pH7 water bath by an electric toothbrushing machine and an electric occlusal force fatigue machine alternatively. Only the buccal surface of every tooth suffered from brushing test. There were 600 000 strokes of occlusal force fatigue test and/or 800 000 cycles of cross-brushing test for every tooth throughout the experiment with 4 intervals. During every interval, the depth at the profile of the cervical defect on each replica of tested tooth was measured and the shape of every defect was recorded with a computerized Seescan image analysis system. The findings suggested that the occlusal force played an important role in the formation of experimental cervical wedge-shaped defects. Occlusal force alone could produce cervical tooth defect in very slow rate. When the occlusal force associated with acid erosion and tooth-brushing abrasion, the depth of cervical defect increased significantly and the wedge-shaped defects were formed. PMID- 8728965 TI - [An experimental study on the effect of operation at one side of the temporomandibular joint on the opposite side]. AB - Condylectomy and menisectomy were performed in left TMJ of 14 rabbits, of them, 7 were treated with periosteal interposition and 7 without. Sixteen weeks after operation, condylar cartilage of the right side were studied and compared with 7 normal rabbits by means of histology and histochemistry. The results show that an operation of high condylectomy and menisectomy with or without interposition on one side of TMJ will result in degenerative changes in the other side (non operated side). The degenerative changes in the rabbits without any interposition are more serious than those with periosteal interposition. PMID- 8728966 TI - [Preventive effect of 10% (NH4)2MoO2F4 solution on artificial root-surface caries in vitro]. AB - The aim of this study is to determine the preventive effect of F-Mo preparation on artificial root-surface caries. 60 premolar teeth extraced for orthodontic purpose were divided into five groups. A window was prepared on every tooth. They were treated by 10% (NH4)2MoO2F4, 38%Ag (NH4)2F, 2% NaF, 7.3% (NH4)2MoO4 and deionized water for 2 minutes and three times. They were then put in the fluid contained cariogenic bacteria for 96 hours to produce root caries. The root surface lesion of every tooth was examined and analysed by micro-radiography, SEM and spectrum. The result showed that the lesion depth of the groups treated by 10% (NH4)2MoO2F4 and 38%Ag (NH4)2F were significantly smaller than the other groups, and the surface of windows are more integrate and smooth, and mean mineral concentration is more higher than those of the other groups. It can be concluded that 10% (NH4)2MoO2F4 solution has greater inhibit effect to the development of root caries, so as the 38% Ag(NH4)2F solution. But the former discolored the tooth much less than the latter. PMID- 8728967 TI - [Effects of occlusal trauma on temporomandibular joints in rabbits: a scanning electron microscopic study]. AB - Six rabbits with unilateral occlusal splints on the mandibular left molars for one month, three months or five months were examined to study the ultrastructure of the surface of the temporomandibular joint. Four rabbits were used as control. The results showed that the surface layer of the disk and articular cartilage were damaged. Severe fibrillation with sheets of fibers, collagen fibrils and collagen bundles inside fissures and craters were observed. These changes were seen in all rabbits with occlusal trauma and also in control rabbits for five months. The findings suggested that occlusal trauma could result in degenerative changes of the temporomandibular joint. PMID- 8728968 TI - [The early treatment of unilateral crossbite]. AB - Fifteen unilateral crossbite patients of primary and mixed dentition with mandibular asymmetry were treated by transverse expansion using removable appliance. Posteroanterior cephalometric analysis was utilized before and after treatment. The results suggested that after the correction of unilateral crossbite, the deviation of midsagittal mandibular total lenth decreased significantly. According to this study, early treatment of unilateral crossbite is indicated. PMID- 8728969 TI - [The anatomic study of levator veli palatini muscle]. AB - The purpose of this study was to obtain more informations about the levator veli palatini (LVP) muscle through the dissection of head and neck cadaver samples in 20 sides of normal adults, 4 sides of normal newborns and 4 sides of cleft palate newborns and histological sections in 6 normal fetuses. The results shown that the middle part of LVP is 7.9 mm in width, 3.4mm in thickness and 29mm2 on cross section area. There are two angle relationship between LVP and the soft palate, which are 44.5 degrees angle to midline sagittal section inward and 33.0 degrees angle to coronal section forward. The LVP insertion is between the 40% to 70% of the soft palate length and only 25% insertion of LVP is located at the junction area of palatal aponeurosis and fibromuscular component of palatine raphe. Other parts of LVP insertion are merged into the midline fibromuscular component. The muscle has a close relationship with the medial part of the cartilaginous portion of Eustachian tube. The LVP is supplied by the palatal branches of ascending palatine artery and ascending pharyngeal artery, which are into the LVP at or above the level of the hard palate. The LVP nerve branches coming from the pharyngeal plexus pass through the lateral pharyngeal space. The LVP reconstruction in cleft palate repair is discussed. PMID- 8728970 TI - [The use of free grafting of muscle flap or myocutaneous flap of latissimus dorsi in plastic surgery of the maxillofacial region]. PMID- 8728972 TI - [T-cell subsets and lymphocyte proliferation in recurrent oral ulcers]. PMID- 8728971 TI - [Low-dose doxycycline therapy: effect on crevicular fluid collagenase elastase and beta-glucuronidase activity in humans]. PMID- 8728973 TI - [The effect of indolacin in rat periapical lesion and radioimmunoassay determination of prostaglandin E2]. PMID- 8728974 TI - [Effects of gu chi san on cultured human gingival fibroblasts]. AB - The present study demonstrates that Gu Chi San is able to increase the bone density in the clinical and animal tests. This study finds that Gu Chi San has no cytotoxicity on HGFs, and 1g/L Gu Chi San extracts can stimulate the mitosis and enhance the DNA synthesis of HGFs. The results indicate that certain substances are present in Gu Chi San. PMID- 8728975 TI - [Relationship between kidney insufficiency and some endocrine hormones in periodontitis patients]. PMID- 8728976 TI - [The management and clinical effects of Herbst appliance]. PMID- 8728977 TI - [Development and current status of dental implantation abroad]. PMID- 8728978 TI - [Biofeedback in speech therapy after cleft palate surgery]. PMID- 8728979 TI - DNA fingerprinting reveals female preference for male parental care in Savannah Sparrows. AB - According to sexual selection theory, females choose mates to ensure access to high quality resources, male parental care, or good genes. This last hypothesis has been hotly debated on both theoretical and empirical grounds. In contrast, female preference for male parental care has received less attention, primarily because the potential benefits of paternal effort seem obvious. The fitness relations are less clear in double-brood species, however, because females can base mating decisions on their prior experience with male parental care. Here, the extent of male parental care delivered to first-brood offspring may indicate male genetic quality and/or be the target of female manipulation via her subsequent mating fidelity. In Savannah Sparrows (Passerculus sandwichensis), multi-locus DNA fingerprinting of 203 adults and young revealed substantial female infidelity in first and second broods: overall, 24 of 80 first-brood young (30%) and 13 of 80 second-brood young (16.3%) were the product of extra-pair fertilizations. Among 12 females altering fidelity between broods, absolute male feeding rate to first-brood offspring was a strong, positive predictor of change in female fidelity. Because the extent of male parental care reflects a male's viability, the data support a 'good-genes' interpretation. PMID- 8728980 TI - Are critical phenomena relevant to large-scale evolution? AB - Recent theoretical studies, based on the theory of self-organized critical systems, seem to suggest that the dynamical patterns of macroevolution could belong to such class of critical phenomena. Two basic approaches have been proposed: the Kauffman-Johnsen model (based on the use of coupled fitness landscapes) and the Bak-Sneppen model. Both are reviewed here. These models are oversimplified pictures of biological evolution, but the (possible) validity of them is based on the concept of universality, i.e. that apparently very different systems sharing some few common properties should also behave in a very similar way. In this paper we explore the current evidence from the fossil record, showing that some properties that are suggestive of critical dynamics would also be the result of random phenomema. Some general properties of the large-scale pattern of evolution, which should be reproduced by these models, are discussed. PMID- 8728981 TI - Image blur as a pictorial depth cue. AB - A range of cues are already known to mediate depth perception in pictures and have been exploited by artists in drawings and paintings. Modern images are commonly generated by photographic or video equipment, and these images contain a depth cue that cannot be found in artistic depictions of natural scenes: different image regions are often blurred by different amounts, because of depth of focus limitations. Demonstrations presented here show that this selective image blur also acts as a pictorial depth cue, even when other pictorial cues are removed. Experimental data indicate that the degree of blur at borders between blurred and sharp image regions is used by the visual system to establish the depth ordering of different regions. Selective image blur is thus a potentially useful addition to computer-generated and cartoon images to enhance the impression of depth they convey. It may well also contribute to depth perception in natural retinal images, because the depth of focus of the human eye is limited. PMID- 8728982 TI - Neocortex size and behavioural ecology in primates. AB - The neocortex is widely held to have been the focus of mammalian brain evolution, but what selection pressures explain the observed diversity in its size and structure? Among primates, comparative studies suggest that neocortical evolution is related to the cognitive demands of sociality, and here I confirm that neocortex size and social group size are positively correlated once phylogenetic associations and overall brain size are taken into account. This association holds within haplorhine but not strepsirhine primates. In addition, the neocortex is larger in diurnal than in nocturnal primates, and among diurnal haplorhines its size is positively correlated with the degree of frugivory. These ecological correlates reflect the diverse sensory-cognitive functions of the neocortex. PMID- 8728983 TI - Developmental switching in the parasitic nematode Strongyloides ratti. AB - Strongyloides ratti is a nematode parasite of rats. It is able to undergo two types of development outside the host: heterogonic (free-living adults and sexual reproduction) and homogonic (direct larval development). Homogonic development has a number of similarities with the development of the dauer stage of free living nematodes, including Caenorhabditis elegans. Using isofemale lines of the parasite, factors that control this developmental choice have been investigated. Isofemale lines can be selected for both heterogonic and homogonic development, but are still able to respond to environmental conditions. By using temperature shift experiments it has been possible to determine when larvae become developmentally committed. All larvae are developmentally committed after 24 h at 19 degrees C. PMID- 8728984 TI - Long, polymorphic microsatellites in simple organisms. AB - We have examined the phylogenetic distribution of the longest, perfect microsatellites in GenBank. Despite the large contributions of model higher eukaryotic organisms to GenBank, the selective cloning of long microsatellites from these organisms as genetic markers, and the relative lack of concentration on the microsatellites in lower eukaryotes and prokaryotes, we found that simple organisms, defined here as slime molds, fungi, protists, prokaryotes, viruses, organelles and plasmids, contributed 78 of the 375 examined sequences. These 78 simple-organism microsatellites are characterized predominantly by trinucleotide repeats, nearly half of which lie in exons, and in general show a bias towards A+T rich motifs. Simple-organism microsatellites represented more than once in GenBank displayed length polymorphisms when independent clones were compared. These facts collectively raise speculation as to the role of these 'junk' sequences in such highly economical genomes, especially when precise changes in long microsatellites are known to regulate critical virulence factors in several prokaryotes. Regardless of their biological significance, simple-organism microsatellites may provide a general source of molecular markers to track disease outbreaks and the evolution of microorganisms in unprecedented detail. PMID- 8728985 TI - Constraints and normalized measures for cytonuclear disequilibria. AB - The full bounds are derived for cytonuclear disequilibria in two-locus systems with an arbitrary number of alleles at the cytoplasmic and nuclear markers. The associated marginal frequencies constrain the nonrandom associations between cytoplasmic alleles and nuclear genotypes in the same way that the allele frequencies constrain the linkage disequilibrium between two nuclear loci. Additional constraints are imposed on the nonrandom associations between cytoplasmic and nuclear alleles, however, by the marginal frequencies of nuclear genotypes carrying either two or no copies of the associated nuclear allele. These bounds are analysed and used to define normalized measures of cytonuclear disequilibria, whose practical utility is illustrated through applications to two sets of recent nuclear-mitochondrial data. PMID- 8728986 TI - RAPD analysis of systematic relationships among the Cervidae. AB - We investigated the possible application of RAPD (Random Amplified Polymorphic DNA) analysis to the study of the systematic relationships of five cervid taxa. Amplifications with eight different primers gave reproducible electrophoretic patterns which could be regarded as a data-set consisting of monomorphic and polymorphic characters. Some of these characters are species- and subspecies specific. Band-sharing analysis and numerical taxonomy methods allowed us to generate a phenetic tree. Our results point out new possible systematic considerations within the examined taxa. PMID- 8728987 TI - Biogeography and taxonomy of Apodemus sylvaticus (the woodmouse) in the Tyrrhenian region: enzymatic variations and mitochondrial DNA restriction pattern analysis. AB - In the western Mediterranean area, the taxonomic status of the various forms of Apodemus sylvaticus is quite unclear. Moreover, though anthropogenic, the origins of the island populations remain unknown in geographical terms. In order to examine the level of genetic relatedness of insular and continental woodmice, 258 animals were caught in 24 localities distributed in Belgium, France, mainland Italy, Sardinia, Corsica and Elba. Electrophoresis of 33 allozymes and mtDNA restriction fragments were performed and a UPGMA dendrogram built from the indices of genetic divergence. The dendrogram based on restriction patterns shows two main groups: 'Tyrrhenian', comprising all the Italian and Corsican animals and 'North-western', corresponding to all the other mice trapped from the Pyrenees to Belgium. Since all the Tyrrhenian mice are similar and well isolated from their relatives living on the western edge of the Alpine chain, they must share a common origin. The insular populations are consequently derived from peninsular Italian ones. From a taxonomic point of view and taking the priority rules into account, we have to invalidate A. s. clanceyi Harrison, 1948 and to consider the Tyrrhenian woodmice as belonging to A. s. milleri de Beaux, 1926, whereas the North-western ones must be referred to as the nominal supspecies. As far as the Elban woodmouse is concerned, at the moment we prefer to keep its present subspecific status because we only studied one animal. PMID- 8728988 TI - Phenotypic and functional properties of murine gamma delta T cell clones derived from malaria immunized, alpha beta T cell-deficient mice. AB - Six murine T cell clones expressing gamma delta TCR were generated from malaria immunized, alpha beta T cell-deficient mice. Phenotypic characterization of these clones has revealed that, in contrast to conventional alpha beta T cells, there is a considerable degree of heterogeneity among these gamma delta clones with regard to their surface markers and their lymphokine profile. One clone was found to display significant anti-parasite activity in vivo upon adoptive transfer. We attempted to determine whether the protective clone differs in one or more key characteristics from the non-protective clones. Although no obvious pattern peculiar to the protective gamma delta clone was observed, it appears that more than one parameter may, in combination, define a distinct protective phenotype, and thus explain the functional difference between the protective and non protective gamma delta clones. PMID- 8728989 TI - The human anion transport protein, band 3, contains a CD36-like binding domain for Plasmodium falciparum-infected erythrocytes. AB - Epitope mapping of a murine monoclonal antibody (mAb), 5H12, prepared against live Plasmodium falciparum-infected red blood cells indicated that the epitope consisted of amino acid residues 474-487 of the human anion transport protein, band 3. mAb 5H12 enhanced cytoadherence, but inhibited the CD36-like mediated rosetting. A synthetic peptide based on the sequence of the epitope (FSFCETNGLE) blocked both rosetting and cytoadherence, suggesting that this amino acid sequence may form the CD36-like receptor. The CD36-like region of band 3 was antigenically distinct from platelet or endothelial CD36. PMID- 8728990 TI - The course of Plasmodium berghei, P. chabaudi and P. yoelii infections in beta thalassaemic mice. AB - In order to study the effects of acclimatization of Plasmodium in beta thalassaemic mice, we used a mouse model of beta-thalassaemia (DBA/2J/beta thal/beta-thal), similar to that observed in humans. We acclimatized 3 rodent malarias (P. berghei, P. chabaudi and P. yoelii) in DBA/2J and DBA/2J/beta-thal mice lines, by 4 intraperitoneal serial transfers. All 3 rodent malarias developed in red blood cells of beta-thalassaemic mice without losing their virulence. There was no delay in infection and peaks of parasitaemia were similar in beta-thalassaemic and normal mice. The mortality occurred earlier in beta thalassaemic mice than in control mice for P. berghei and P. chabaudi. The difference was more pronounced for P. yoelii NS where normal mice did not die. These results could be explained by a failure of erythropoiesis in beta thalassaemic mice, which are unable to compensate for the destruction of red blood cells by the parasites, and the mice died of anaemia. Ultrastructural examination of the rodent malaria parasites in beta-thalassaemic RBC showed a normal development even in the presence of Heinz bodies. In conclusion, no effective protection against malaria was provided by the beta-thalassaemia in this mouse model. PMID- 8728991 TI - During canine viscero-cutaneous leishmaniasis the anti-Hsp70 antibodies are specifically elicited by the parasite protein. AB - A Leishmania infantum cDNA library was screened with sera from dogs with viscero cutaneous leishmaniasis. Sequence analysis of a positive clone isolated from the library revealed that it coded for the carboxyl-terminal region of a member of the 70-kDa heat-shock protein family. The full-length sequence of the L. infantum hsp70 gene was determined after isolation of genomic clones. This protein shows a high degree of sequence conservation with the homologous protein from other organisms. To test its antigenicity a recombinant Hsp70 protein fused to the maltose-binding protein was produced in Escherichia coli using the expression vector pMAL-cRI. By FAST-ELISA assays it was observed that while the complete recombinant protein was recognized by 100% of the sera, the 20 carboxyl-terminal amino acids of the protein were only recognized by 30% of those sera. Thus, although a B-cell epitope must be present within the carboxyl terminal end of the protein other antigenic determinant(s) must reside out of this region. The analysis of the cross-reactivity with mouse Hsp70 by Western blotting strongly suggests that the anti-Hsp70 antibodies generated by infection with L. infantum are directed at specific determinants of the L. infantum Hsp70. Thus, our results indicate that anti-Hsp70 autoantibodies are not induced during Leishmania infection. PMID- 8728992 TI - Patterns of haemozoin accumulation in tissue. AB - A sensitive fluorometric method for assaying malarial pigment, haemozoin, has been developed and used to determine the haemozoin content of blood and tissue samples. Plasmodium falciparum rings and trophozoites were found to contain 23 and 339 ng haemozoin/10(6) parasitized red blood cells (PRBCs), respectively. Unsynchronized Plasmodium berghei NK65 or ANKA parasites from infected mice contained 27 and 61 ng haemozoin/10(6) PRBCs, respectively. An exponential accumulation of haemozoin within 18 days after infection was demonstrated in liver and spleen tissue, representing up to 0.2% of the tissue by wet weight by day 18. Histology indicated that the accumulation occurred predominantly in the tissue monocytes. In the brain, the levels of haemozoin after 8 days of infection were considerably lower than they were in the liver or spleen, and most of the pigment appeared to be that present inside parasitized red blood cells. CBA/Ca mice infected with P. berghei ANKA (a cerebral malaria model) had significantly higher amounts of haemozoin in the brain than did ICR mice infected with P. berghei NK65. Thus, haemozoin levels in tissue increase with the duration of infection, and its presence may be associated with cerebral pathology. PMID- 8728993 TI - Neutral glycolipids of Schistosoma mansoni as feasible antigens in the detection of schistosomiasis. AB - The neutral glycolipid fraction from mouse-propagated, Schistosoma mansoni adult worms has been investigated as to its chromatographic and antigenic properties, and whether it fulfills the serodiagnostic antigen requirements of sensitivity and specificity in the detection of schistosomiasis. Serological analyses were performed by thin-layer chromatography immunostaining and ELISA. In the acute phase form of mouse schistosomiasis, the kinetics of development of neutral glycolipid-specific antibody levels was correlated with the intensity of the initial infection and the response was dominated by IgG, as represented by the subclass IgG1. With the experimental animal helminthiases screened, glycolipid antigenicity fulfilled the fundamental traits for a serodiagnostic reagent. In the chronic-phase form of human schistosomiasis mansoni, neutral glycolipid specific antibody levels were not correlated with the intensity of infection, as estimated from the faecal content of parasite eggs, whilst the isotypic response was dominated by IgM and IgG, the latter represented primarily by IgG1 and secondarily by IgG3. With other human helminthiases, glycolipid antigenicity was incomplete, in that, the specificity was only partially fulfilled. The reason for this incomplete specificity has been clarified, in part, by the detection of cryptic schistosomiasis infections in the cohorts of African patient sera examined. PMID- 8728994 TI - Quinacrine-resistant Giardia duodenalis. AB - Quinacrine resistance has been induced in 3 Giardia duodenalis laboratory stocks and 4 lines resistant to other drugs. The quinacrine-resistant lines were maintained in normally lethal levels of 5 microM quinacrine and some lines are viable in 20 microM levels of the drug. Fluorescence studies indicated that quinacrine was taken up by sensitive cells but was actively excluded from resistant trophozoites. The nuclei were not a site of drug accumulation and no specific fluorescence in the trophozoite could be attributed to any structure. Blebs of concentrated drug appeared prior to disintegration of the membrane of drug-sensitive trophozoites exposed to drug overnight. Parasite lines already resistant to furazolidone adapted more readily to quinacrine exposure than drug sensitive stocks. This multiple drug resistant phenotype was not as marked with metronidazole-resistant lines. PMID- 8728995 TI - Susceptibility of Brugia malayi and Onchocerca lienalis microfilariae to nitric oxide and hydrogen peroxide in cell-free culture and from IFN gamma-activated macrophages. AB - The susceptibility of Brugia malayi and Onchocerca lienalis microfilariae to H2O2 and NO either in cell-free culture or from IFN gamma-activated macrophages was examined. In cell-free culture, O. lienalis microfilariae were highly susceptible to H2O2 induced toxicity, exhibiting rapid reductions in motility and viability. The addition of exogenous catalase abrogated H2O2-induced killing. In contrast, B. malayi microfilariae were relatively resistant to H2O2, with concentrations as high as 50 microM having no effect on motility or viability. On exposure to NO, both species showed reductions in motility within 5-30 min, but longer was required to see effects on the viability of microfilariae. Parasites incubated with IFN gamma-activated macrophages also exhibited marked reductions in motility and viability. In cultures with B. malayi and activated macrophages, inhibition of these effects was achieved by the addition of either L-NMMA, to abolish NO production, or neutralizing anti-TNF alpha antibodies. Attempts to inhibit parasite killing by the addition of catalase to macrophage cultures were ineffective. The results of this study show that B. malayi and O. lienalis microfilariae have different susceptibility to H2O2, but are equally affected by exposure to NO. Moreover both species are killed by IFN gamma-activated macrophages and in the case of B. malayi, killing is dependent on the generation of NO via TNF alpha. PMID- 8728996 TI - Eosinophils are the major effector cells of immunity to microfilariae in a mouse model of onchocerciasis. AB - Mice inoculated with microfilariae of the filarial nematode Onchocerca lienalis clear their parasites over a period of 3-4 months and are highly resistant to re infection. We have investigated the comparative roles of the eosinophil, macrophage and neutrophil in effecting this parasite clearance, employing agents specifically to perturb cell function in vivo. Using the anti-IL-5 monoclonal antibody TRFK-5, we show that eosinophils are of primary importance in effecting resistance to re-infection. Ablation of macrophages (with carbon) and neutrophils (with the monoclonal antibody NIMP-R14) had no effect on parasite clearance following re-infection. Neutralization of these 3 cell types during a primary infection showed that while the removal of both eosinophils and macrophages caused a small but significant delay in parasite clearance, the depletion of neutrophils had no effect. This report describes the first direct evidence for eosinophil-mediated killing of microfilariae in vivo, and is consistent with Th-2 cell responses previously described in this model. PMID- 8728997 TI - Cloning and expression of DiT33 from Dirofilaria immitis: a specific and early marker of heartworm infection. AB - Dirofilaria immitis is an important filarial parasite of dogs and cats, and a useful model for human filariasis. Current diagnostic tests for heartworm infection in animals rely on the presence of fecund female worms (usually found 6.5 months post-infection or later) and therefore fail to detect pre-patent infections. Putative pepsin inhibitors from 2 filarial parasites of humans namely Onchocerca volvulus (Ov33, Oc3.6, OvD5B) and Brugia malayi (Bm33), have been shown to be useful in diagnosis of onchocerciasis and lymphatic filariasis, respectively. Previous studies have suggested that a homologue exists in D. immitis (DiT33), which may have potential in diagnosis of heartworm infection. In this study, the isolation and characterization of a cDNA clone encoding DiT33 is described. This cDNA contains 12 bases of the nematode-specific 22 nucleotide spliced leader sequence and encodes a 26.4 kDa-protein with a high level of similarity (87-89%) to other filarial members of the family. DiT33 was over expressed in E. coli as a fusion with the maltose-binding protein and serological analysis was performed using a panel of clinically defined dog sera. The findings of this study indicate that DiT33 is a promising antigen for the early detection of D. immitis and may be a valuable tool in the control and management of heartworm infection. PMID- 8728998 TI - Dissociation of early and late protective immunity to the nematode Nippostrongylus brasiliensis in Brown Norway and Fischer-344 rats. AB - Worm expulsion of, and IgE and interferon (IFN)-gamma responses to, Nippostrongylus brasiliensis were studied in 2 rat strains, Brown Norway (BN) and Fischer (F)-344. BN rats expelled the majority of worms by day 14 post-infection (p.i.) with approximately 6% of worms surviving for at least 3 weeks. In F-344 rats, worm expulsion was delayed by 2 days relative to that in BN, while the numbers of residual worms were significantly fewer than in BN, suggesting that different immune mechanisms are involved in early and late phases of immunity. Total serum IgE, as well as in vitro IgE production by mesenteric lymph node (MLN) cells, was increased 2 weeks p.i., the levels being markedly higher in BN than in F-344 rats. Serum rat mast cell protease II was also increased more significantly in BN than in F-344 rats. In contrast, production of IgG2a and IFN gamma by MLN and spleen cells was found to be higher in F-344 than in BN rats. These results indicate that the early worm expulsion is correlated with the host IgE and mast cell responsiveness, whereas the persistence of infection in the late period may be controlled by different immune mechanisms. PMID- 8728999 TI - Establishment, survival and site selection of the cestode Eubothrium crassum in brown trout, Salmo trutta. AB - Population dynamics, site selection, growth and maturation of the cestode Eubothrium crassum in a natural population of Salmo trutta in a small lake were studied over a period of 1 year, the life-span of a cohort in fish. Infection of fish commenced in spring but peaked in July. Small, plerocerciform parasites initially located in the intestine, but then some moved into the pyloric caecae whilst others, the majority in heavy infections, were lost from the fish causing a fall in abundance from 460 to 10 over 2 months. This mortality was density dependent. Initially, parasites were distributed more evenly throughout the caecae but as time increased larger parasites were found preferentially in the anterior caecae before moving back into the intestine when gravid, preparatory to being lost in the following summer. Only a small proportion of the infrapopulation became gravid. Although the proportion of caecae occupied was initially density dependent, by the time of maturation several preferred anterior caecae remained unoccupied and mean intensity always exceeded unity. Neither growth nor maturation was affected by intraspecific competition. It was concluded that caecal availability did not set a limit or threshold of infrapopulation density, and in this respect E. crassum-S. trutta differed from some acanthocephalan-fish systems but was similar to others. Heavy infection followed by heavy mortality appeared to be typical of this parasite-host system in other localities, and of several other cestode-fish systems. The implications of this for population regulation are discussed. PMID- 8729000 TI - Matrilysin: an epithelial matrix metalloproteinase with potentially novel functions. AB - Matrilysin (MAT) is a member of the matrix metalloproteinase (MMP) family which is believed to degrade components of the extracellular matrix (ECM) during processes of tissue remodeling. Although MAT is similar to the stromelysins in its substrate specificity, and to interstitial collagenase in the crystal structure of its catalytic domain, this enzyme is unique in that it lacks the carboxy-terminal segments encoded by other MMP genes. Characterization of the human MAT gene has revealed that the promoter region contains typical MMP promoter elements such as AP-1 and PEA3, which mediate responsiveness to growth factors, oncogenes, and phorbol esters. Activated recombinant forms of human MAT cleave ECM and basement membrane proteins such as fibronectin, collagen type IV, laminin, and particularly elastin, entactin, and cartilage proteoglycan aggregates. Furthermore, MAT appears to mediate the proteolytic processing of other molecules (e.g. tumor necrosis factor alpha precursor, urokinase plasminogen activator). MAT is expressed in a variety of tumors ranging from adenomas to carcinomas and adenocarcinomas of the breast, colon, prostate, stomach, upper aerodigestive tract, lung, and skin, where it may be involved in tumor formation as well as the tissue degradation which accompanies tumor cell extravasation. Localization of MAT mRNA and protein to the tumor cells is unusual in that the majority of MMPs are produced in the stroma. This distinctive tissue restricted pattern of MAT expression is a recapitulation of the expression pattern in normal human tissue, where MAT protein localizes to secretory and ductal epithelium in the endometrium and in various exocrine glands. In the mouse, high constitutive levels of MAT mRNA are found in epithelial cells in the uterus, small intestine, and extra-testicular ducts. Taken together, these findings suggest that MAT may have a specific role in normal gland and organ function, a possibility which can be explored further by the genetic manipulation of MAT levels in vivo. PMID- 8729001 TI - A possible mechanism for initiation of lipid peroxidation by ascorbate in rat liver microsomes. AB - The mechanism by which lipid peroxidation progresses has been known for years, but there is disagreement regarding the mode of its initiation. The aim of this study was to examine: (a) the role of endogenous iron in the initiation of ascorbate-induced lipid peroxidation in microsomal and liposomal membranes; (b) the role of oxygen-free radicals in this process; and (c) the redox state of ascorbate during the course of lipid peroxidation. Ascorbate-induced lipid peroxidation was assessed by measuring hydroperoxide and thiobarbituric acid reactive substances (TBARS) formation in membranes after incubation in Tris-HCl buffer (pH 7.4) for 15 min. To confirm the role of endogenous iron and oxygen free radicals, the effect of iron chelating agents (EDTA and thiourea) and radical scavengers (benzoate, mannitol, catalase and SOD) on lipid peroxidation was examined. Spectrophotometric measurements and ESR spectra have made it possible to determine ascorbate concentration and its redox state. Ascorbate promoted lipid peroxidation in both rat liver microsomes and liposomes without addition of exogenous iron. Iron chelating agents such as EDTA and thiourea inhibited lipid peroxidation, while SOD, catalase, mannitol and benzoate had no effect. The addition of 5 microM Fe2+ (or Fe3+) to the incubation mixture did not significantly alter hydroperoxide production, but that of TBARS was increased. Lipid peroxidation significantly altered the fatty acid profile in microsomes and liposomes, the most affected being the C20:4 and C22:6 species. Ascorbate in Tris HCl buffer (pH 7.4) autoxidized very slowly. Its oxidation was catalyzed by Fe3+ ions at a rate determined by incubation time and iron concentration. In contrast, no ascorbate oxidation occurred in the presence of microsomes when lipid peroxidation was proceeding at a maximal rate. Under these conditions a typical ascorbyl radical ESR spectrum signal greater than that arising from ascorbate alone was obtained and the magnitude of this signal was unchanged by variations of microsome or ascorbate concentrations. A ferrous ion ascorbyl radical complex was responsible for this signal. These results suggest that an ascorbate microsomal iron complex is responsible for the initiation of lipid peroxidation, and that during this process ascorbate remains in its reduced form. PMID- 8729002 TI - Cholesterol-loading of peripheral tissues alters the interconversion of high density lipoprotein subfractions in rabbits. AB - High density lipoprotein (HDL) has been implicated in the process of reverse cholesterol transport,by which surplus cholesterol is removed from peripheral tissues and transported to the liver for excretion. It has been suggested that some subfractions of HDL may have a particular role in this process, though the underlying mechanism remains unclear. The present study was aimed at investigating the role of specific subfractions of HDL in reverse cholesterol transport. The interconversion of HDL subfractions in normal and cholesterol loaded rabbits was studied in vivo. Rabbit HDL was separated by heparin-Sepharose affinity chromatography into six subfractions (HDL(I)-HDL(VI)), which were progressively enriched with apolipoprotein E (apo E), and varied in diameter and composition. Total HDL and its subfractions were individually labelled with 14C sucrose and injected in the rabbits. When rabbits which were not acutely loaded with [3H]cholesterol were injected with 14C-HDL(I), 70% of the label remained in this fraction while less than 5% was recovered in HDL(VI), containing the largest particles and those most enriched in apo E. No label was detectable in the liver of these animals. In rabbits which had received a prior loading of cholesterol, an average of only 18.3% of the 14C label was present in HDL(I) while approx. 40% of the label was recovered in HDL(VI). On average, 5.1% of the total 14C injected in these rabbits was recovered in the liver. It is concluded that two alternative routes for reverse cholesterol transport may be operative. While a continuous cholesterol-clearance route may be provided by particles of HDL of intermediate size, another route may be operative for clearance of excess cholesterol loaded into peripheral endothelial cells. PMID- 8729003 TI - Characterization and purification of fucosyltransferases from the cytosol of rat colon. AB - The occurrence and baseline characteristics of fucosyltransferases (alpha-1,2, alpha-1,3 and alpha-1,4) in the cytosol (soluble) and pellet (membrane-bound) of rat colon have been studied since the fucosylation process is known to alter in colon pathology. All enzymes studied in the colon pellet had higher activity when compared to the cytosol. The colon pellet alpha-1,3 fucosyltransferase preferred desialylated alpha 1-acid glycoprotein as acceptor substrate. Both soluble and membrane-bound enzymes, alpha-1,2 and alpha-1,3 fucosyltransferases, required Mn2+, Mg2+ and Ca2+ for maximum activity but were inactivated by Cu2+ ions. Both soluble alpha-1,2 and alpha-1,3 fucosyltransferases showed optimal activity at pH 6.0, whereas the optimum for their membrane-bound activities were at pH 5.8 and 6.2, respectively. Furthermore, a soluble alpha-1,3 fucosyltransferase from rat colon was purified and during purification the co-presence of alpha-1,3/4 fucosyltransferase was detected. The acceptor of preference for the purified soluble alpha-1,3 fucosyltransferase was desialylated glycoprotein while low molecular weight substrates were poor acceptors. Both the purified fucosyltransferases were inhibited by N-ethylmaleimide. The M(r) values determined by SDS-PAGE electrophoresis of alpha-1,3/4 fucosyltransferase and of alpha-1,3 fucosyltransferase were 68,780 and 40,680 respectively. In conclusion, based on their properties, the purified soluble colon alpha-1,3 fucosyltransferase appeared to be of plasma-type (or FT-I) while the soluble alpha-1,3/4 fucosyltransferase corresponded to Lewis-type or FT-III. PMID- 8729004 TI - Truncated erythropoietin receptor in a murine erythroleukemia cell line. AB - The Friend spleen focus forming virus produces a 55 kDa envelope glycoprotein which associates with the erythropoietin receptor. We compared the erythropoietin receptor in Friend virus transformed murine erythroleukemic F4N and 707 cell lines with the J2E erythroid line generated by the J2 retrovirus. Reverse transcriptase PCR was used to determine transcript size. Erythropoietin receptor cDNAs were then sequenced and protein products analysed by Western blotting and immunoprecipitation. We show here that the F4N murine erythroleukemic cell line had an enlarged erythropoietin receptor mRNA. In contrast, the 707 and J2E cell line had normal sized transcripts for the receptor. Sequence analysis of the receptor in F4N cells revealed that introns which separate the exons coding for the cytoplasmic domain of the receptor were retained in these transcripts. As a consequence, a premature stop codon had been introduced, leaving only four amino acids in the intracellular portion of the receptor molecule. The normal erythropoietin receptor is approx. 66-70 kDa, but immunoprecipitation of [35S]methionine/cysteine labelled cell lysates with an antibody to the amino terminus of the erythropoietin receptor identified a truncated 37 kDa protein in F4N cells. Despite the severe carboxy-terminal truncation of the erythropoietin receptor, F4N cells continued to proliferate like the other murine erythroleukemia cell lines. This study shows that failure to remove introns from the erythropoietin receptor mRNA in F4N cells has resulted in the production of a smaller protein with virtually no cytoplasmic domain. PMID- 8729005 TI - Counter modulation of adipocyte mitochondrial processes by insulin and S oxalylglutathione. AB - Oxalyl thiolesters, a group of putative intracellular regulators, have been shown to be in vitro inhibitors of some cytosolic enzymes which are stimulated by insulin. In this study, the effects of insulin and oxalyl thiolesters on pyruvate dehydrogenase, beta-oxidation, and acyl-CoA hydrolase activities in mitochondria from rat epididymal adipocytes are compared. Using glutathione, CoASH, cysteine, and cysteamine as thiol sources, oxalyl thioesters were synthesized, purified, and quantitated. Mitochondria were isolated from rat epididymal adipocytes, some of which were incubated with or without insulin. Mitochondrial activities were determined by radioisotopic assay subsequent to control, insulin, or oxalyl thiolester incubation. Under the conditions used in this study, pyruvate dehydrogenase activity was increased 28% subsequent to 10-min incubation of adipocytes with 400 microU/ml insulin; in contrast, preincubation of adipocyte mitochondria with S-oxalylglutathione resulted in a dose-dependent 11-19% inhibition of pyruvate dehydrogenase. S-oxalylglutathione also attenuated the spermine-induced activation of pyruvate dehydrogenase. Insulin treatment resulted in a small but significant increase in beta-oxidation of palmitic acid while 100 microM S-oxalylglutathione mediated a 40% decrease in palmitate oxidation. Palmitoyl-CoA hydrolase activity was decreased 14% by insulin treatment; however, S-oxalylglutathione caused a 14-50% increase in hydrolase activity. The other oxalyl thiolesters were not as effective or as consistent as S-oxalylglutathione in modulation of the mitochondrial activities; free thiols and oxalic acid did not modulate the activities. In summary, pyruvate dehydrogenase, palmitate beta oxidation, and palmitoyl-CoA hydrolase activities in adipocyte mitochondria were modulated in approximately equal but opposite directions by insulin and S oxalylglutathione. These findings support the suggestion that oxalyl thiolesters may function as an intracellular signal recruited to return insulin to normal levels. PMID- 8729006 TI - Bile acid-induced alterations of mucin production in differentiated human colon cancer cell lines. AB - Damage to the gastrointestinal tract mucous layer may render underlying cells susceptible to intraluminal toxins or carcinogens. Our aim was to determine the effect of bile acids on mucin, the primary constituent of mucous. Differentiated Caco-2 and HT29 cells were used as models of human colonic epithelial cells. Mucin was measured by [3H]-glucosamine labeling. Short term (30 min) incubations with 1-5 mM unconjugated bile acids or taurodeoxycholic acid induced mucin release relative to bile acid hydrophobicity. Longer incubations were cytotoxic. Long term (7 days) incubation at nontoxic concentrations (0.1 mM) of deoxycholic acid (DC) decreased total mucin by 36 +/- 2% (SEM, P = 0.0003) in differentiated HT29 cells and by 57.2 +/- 2% (P < 0.05) in Caco-2 cells. Tauroursodeoxycholic acid (TUDC) or ursodeoxycholic acid (0.1-0.5 mM) did not alter mucin levels. Simultaneous incubation of 0.1 mM DC and 0.1-0.5 mM TUDC or 2.5 mM TDC and TUDC did not change mucin levels. Differentiated HT29 and Caco-2 cells contained high levels of intestinal mucin MUC3 mRNA while undifferentiated HT29 cells did not possess a MUC3 message. Deoxycholic acid (0.1 mM) did not alter the MUC3 mRNA level. Neither cell type showed detectable expression of intestinal MUC2 or gastric MUC6. Thus, cytotoxic concentrations of bile acids induce mucin release, presumably due to detergent effects. Nontoxic concentrations of DC reduce mucin levels in differentiated enterocyte-like cells, which can be prevented by coincubation with TUDC. The bile acid-induced alterations in mucin production by enterocytes observed in vitro may influence intestinal cytoprotection in vivo. PMID- 8729007 TI - Importance of specific amino acids in protein binding sites for heparin and heparan sulfate. AB - Heparin and heparan sulfate bind a variety of proteins and peptides to regulate many biological activities. Past studies have examined a limited number of established heparin binding sites and have focused on basic amino acids when modeling binding site structural motifs. This study examines the prevalence of individual amino acids in peptides binding to heparin or heparan sulfate. A 7-mer random peptide library was synthesized using the 20 common amino acids. This 7 mer library was affinity separated using both heparin and heparan sulfate Sepharose. Bound peptide populations were eluted with a salt step gradient (pH 7) and analysed for amino acid composition. Peptides released from heparin-Sepharose by 0.3 M NaCl were enriched in arginine, lysine, glycine and serine; and depleted in methionine and phenylalanine. In contrast, peptides released from heparan sulfate-Sepharose were enriched in arginine, glycine, serine, and proline (at 0.15 M NaCl). These peptides were depleted in histidine, isoleucine, methionine (not detectable) and phenylalanine. In the heparin binding sites of proteins, which have been published, the enriched amino acids were arginine, lysine and tyrosine. Depleted amino acids include aspartic acid, glutamic acid, glutamine, alanine, glycine, phenylalanine, serine, threonine and valine. This study demonstrates that heparin and heparan sulfate bind different populations of peptide sequences. The differences in amino acid composition indicate that the positive charge density and spacing requirements differ for peptides binding these two glycosaminoglycans. PMID- 8729008 TI - Nuclear transcription factor Oct-1 binds to the 5'-upstream region of CYP1A1 and negatively regulates its expression. AB - The cytochrome P450-dependent monooxygenases, which represent an extended superfamily, catalyze the biotransformation of many endogenous and exogenous substances. One of these hemoproteins, cytochrome P4501A1, is most closely associated with the bioactivation of polycyclic aromatic hydrocarbons such as benzo[a]pyrene, which may play a role in environmental carcinogenesis. A negative regulatory element (NRE) has been localized in the 5'-upstream region of the cytochrome P4501A1 gene (CYP1A1) at -843 to -746 base pairs from the site of transcription. The purpose of this research was to define any interactions of trans-acting proteins with this cis element. Rat liver nuclei were used as the source of trans-acting proteins and a biotinylated NRE-bearing fragment (-782 to 843 bp) from a plasmid which contained the CYP1A1 was prepared by the polymerase chain reaction technique. Gel mobility shift assays were used to demonstrate interactions between this NRE fragment and nuclear proteins. The specific binding to an octamer-containing motif in the 5'-upstream region of CYP1A1 was demonstrated; this was used as a step in the partial purification from rat liver of the transcription factor, Oct-1. Conventional chromatographic procedures and DNA recognition site affinity chromatography were also used. HepG2 human hepatoma cells were transfected with both pMCoLUC+ which contains the luciferase gene as a reporter gene driven by the CYP1A1 promoter (including the NRE), and an Oct-1 expression vector. Luciferase activity/mg protein in the doubly-transfected cells was significantly lower than in cells containing only pMCoLUC+. A nuclear transcription factor Oct-1 interacts with a portion of the NRE of the rat CYP1A1, suppressing the expression of this gene. These findings may help to explain the low level of basal expression of CYP1A1 in mammalian systems. PMID- 8729009 TI - Changes in collagen metabolism in response to endothelin-1: evidence for fibroblast heterogeneity. AB - Endothelin-1 (Et-1) is a 21-amino acid peptide primarily synthesized by endothelial cells. It was originally classified as a potent vasoconstrictor but recent evidence suggests that it also possesses a wide variety of non-vascular actions. It stimulates fibroblast and smooth muscle cell proliferation and it has been shown to stimulate fibroblast collagen metabolism. However, studies on its ability to regulate collagen production remain incomplete, and its effect on post translational processing of procollagen has not been studied. This report details the effect of Et-1 on the rates of procollagen synthesis and degradation in two fibroblast cell lines; human foetal lung (HFL-1) and whole foetal rat fibroblasts (Rat 2). Fibroblast cultures were incubated for 24 hr in the presence or absence of Et-1 before procollagen metabolism was determined by measuring hydroxyproline. Non-collagen metabolism was also determined in these cultures from the uptake of tritiated phenylalanine. Et-1 stimulated procollagen synthesis in HFL-1 fibroblasts and reduced synthesis in Rat 2 cells. The response was dose dependent with the greatest effect at 1.10(-6) M Et-1 for both cell types (155 +/- 6% of control (mean +/- SD, n = 6, P < 0.01) and 61 +/- 4% of control (n = 4, P < 0.01) for HFL-1 and Rat 2 fibroblasts, respectively). Non-collagen protein synthesis was increased to 148 +/- 5% of control (P < 0.05) at 1.10(-6) M Et-1. Non collagen protein synthesis remained unaffected in the HFL-1 fibroblast cultures. Procollagen degradation, expressed as a proportion of total procollagen synthesis, was decreased in HFL-1 fibroblasts (control, 29 +/- 2%; Et-1, 1.10(-6) M; 21 +/- 2%; P < 0.01), and increased in Rat 2 fibroblasts (control 42 +/- 1%; Et-1, 1.10(-6) M; 49 +/- 1%; P < 0.01). Blocking of the EtA receptor for Et-1, using the receptor antagonist-BQ123, abolished the effect of Et-1 on procollagen metabolism in both cell types. These results suggest that different populations of fibroblasts exhibit heterogeneous responses to Et-1. It is concluded that Et-1 may play an important role in the extent and distribution of fibrosis seen in diseases associated with the overproduction of Et-1. PMID- 8729010 TI - Purification and characterization of the alcohol dehydrogenase from a novel strain of Bacillus stearothermophilus growing at 70 degrees C. AB - The biocatalysts isolated from thermophilic microorganisms are the object of ever growing scientific interest for (i) the comprehension of the molecular basis of their thermal tolerance, and (ii) their use in different bio-industrial fields. Here we report the purification and characterization of an alcohol dehydrogenase (designated ADH-hT) from the novel strain LLD-R of Bacillus stearothermophilus which grows at 70 degrees C. ADH-hT was obtained in pure form by anion exchange chromatography and two affinity chromatographies, with a final yield of about 30%. ADH-hT was found to be a tetramer of 37 kDa-subunits, and to have a pI of 4.9. ADH-hT displayed a broad substrate specificity; its activity was highest for aldehydes, and decreased progressively for alcohols and ketones. ADH-hT was endowed with catalytic activity and resistance in the presence of several denaturing agents (organic solvents, detergents, chaotropic agents). ADH-hT shared with ADH 1503 (the alcohol dehydrogenase from B. stearothermophilus strain NCA 1503 which grows at 55 degrees C) the optimal temperature of 65 degrees C, but it was more resistant than ADH 1503 towards heating. In conclusion, due to its stability and broad substrate specificity ADH-hT could be utilized in bio industrial processes. Furthermore, we believe that ADH-hT could represent a good model system for studying the mechanism(s) which proteins exploit to gain heat resistance. PMID- 8729011 TI - Auditing ophthalmology audits. AB - Medical audit is now a well-established part of the routine of ophthalmology departments. Accomplishing a complete audit is a complex task and its effectiveness depends on how it is performed. In order to evaluate the procedure in our department a retrospective analysis of 18 audits was carried out with particular reference to the level, objectives, beneficial effects, and effects on current practice. Our practice was critically analysed using these factors. One audit fulfilled the criteria for a full audit. Eleven (60%) were partial or potential and six (30%) were an attempt in planning an audit. The majority omitted plans for implementing change or targets for future assessment. Seven had educational benefit. There was little effect on patient care, cost-effectiveness or communication. Medical audit can be clinically effective, educational and stimulating. Failure to complete all stages affects the outcome. This study identifies the difficulties involved in performing complete audits and suggests improving the procedure by auditing audits on a regular basis. PMID- 8729012 TI - A survey of corneal graft practice in the United Kingdom. AB - A questionnaire designed to survey current corneal graft practice was sent to 498 consultant ophthalmologists in the United Kingdom. Three hundred and twenty-nine completed questionnaires (66%) were returned. Seventy per cent of these were from consultants who perform corneal grafts, of whom 36% had a specialist interest in corneal surgery. The survey found that most consultants preferred to perform corneal grafts on an inpatient basis with the patient under general anaesthesia. Agreement (> 80%) was also found in the following areas: use of a hand-held trephine, concurrent cataract surgery (if indicated), post-operative immunosuppression, and refraction to assess post-operative astigmatism. There was less agreement on the choice of donor material, use of tissue matching, donor trephine size disparity, suture technique, relative timing of trabeculectomy surgery (if required), the management of intraocular lenses during surgery for pseudophakic bullous keratopathy, timing of discharge after surgery, use of prophylactic acyclovir, management of astigmatism, routine removal of all corneal sutures, and discharge of uncomplicated cases from routine follow-up. PMID- 8729013 TI - Indicator-based audit of cataract surgery in four neighbouring hospitals in East Anglia. AB - To review and compare management and outcomes of patients undergoing cataract surgery in order to improve practice by identifying weaknesses and standardising best practice where appropriate, a concurrent and prospective audit from June to October 1993 was carried out in four neighbouring ophthalmic units in East Anglia. Six hundred and twenty-seven consecutive patients were undergoing cataract surgery in the audit period. The main measures and results were as follows: (1) Patients with visual acuity reduced to 6/60 or less should not wait longer than 3 months from consultation to surgery; 69.5% met standard. (2) Patients with visual acuity reduced to 6/18 or less should not wait longer than 12 months from consultation to surgery; 85.8% met standard. (3) Patients who have had cataract surgery should achieve 6/12 or better corrected visual acuity by 3 months post-operatively; 88.6% met standard. (4) There should be less than 2% sight-threatening complications of surgery; 4.2% suffered sight-threatening complications. (5) There should be no life-threatening complications of surgery; 100% met standard. The audit identified key areas of variation in practice, and analysis of reasons for differences in outcome has led to some changes in the management of patients with cataracts in the four units. PMID- 8729014 TI - Cataract surgery by junior ophthalmologists. AB - The first 50 cases of cataract extraction and lens implant operations performed by seven junior ophthalmologists were retrospectively reviewed to determine whether the surgical outcome of cataract surgery compares with the results of the National Cataract Surgery Survey. Main outcome measures were final visual acuity achieved and the incidences of surgically related complications. In all, 322 case notes were reviewed and full datasets were obtainable in 307 patients (95%). Overall 97% achieved a final visual acuity of 6/12 or better. Twenty per cent had coexisting ocular pathology, 7% had intraoperative complications and 21% had post operative complications. The visual outcome compared acceptably with the results of the National Cataract Surgery Survey in Britain. The overall incidences of surgically related complications were also acceptable but the rate of vitreous loss was higher and post-operative complications tended to be of greater magnitude. PMID- 8729015 TI - Ocular outcome in children born before 32 weeks gestation. AB - Ocular outcome was assessed in a 2 year cohort of all children born before 32 weeks gestation within a geographically defined population of approximately 3 million. Five hundred and sixty-five children were born within the study period; 558 children (98.8% of the study group) were assessed at approximately 2 years old. There was a high incidence of abnormalities, particularly strabismus (70, 12.5%), cortical visual loss (7, 1.3%), sequelae of retinopathy of prematurity (16, 2.9%) and other significant refractive errors (69, 12.7%). These data provide a recent estimate of gestational age-specific prognosis and suggest that follow-up yields a significant proportion of children with visual abnormalities who could benefit from treatment. PMID- 8729016 TI - An audit of the 1995 Royal College of Ophthalmologists guidelines for screening for retinopathy of prematurity applied retrospectively in one regional neonatal intensive care unit. AB - To test the effectiveness of the 1995 Royal College of Ophthalmologists (RCO) guidelines for screening for retinopathy of prematurity (ROP) in one regional neonatal intensive care unit, an analysis was carried out of screening examination results over a 4 year period, retrospectively applying the rules of the 1995 RCO guidelines. Four hundred and forty-eight examinations would have been performed in 258 infants, significantly fewer than the 764 examinations which had been performed using our existing screening protocol. Threshold ROP developed in 36 infants, and would have been detected promptly in every case. No case of threshold ROP developed in infants of birth weight greater than 1250 g. The 1995 RCO guidelines for ROP screening provide an effective and efficient means of detecting treatable ROP. The need to include infants of birth weight > 1250 g should be reviewed at a future date, following a period of further national data collection. PMID- 8729017 TI - Glaucoma screening with oculokinetic perimetry in general practice: is its specificity acceptable? AB - The main aim of the study was to estimate the specificity of oculokinetic perimetry (OKP) for glaucoma case-finding in general practice. The visual fields of 185 consecutive patients aged 40 years or more were screened with OKP in one general practice. Test specificity was 94%. The OKP test was abnormal in 17 patients. At follow-up 6 of these patients had glaucoma or were glaucoma suspects. Three of the remaining 11 patients had cataract. Eight OKP-positive patients had no eye disease. In 96 of the patients results of OKP were compared with Bjerrum campimetry, which revealed one possibly false-negative OKP test. Ten patients with previously diagnosed glaucoma were also examined with OKP. All 10 patients were identified by the OKP test. In 80 of the patients OKP was performed twice, supervised by two different observers. The proportion of agreement for normal tests was very high: 0.96 (95% CI: 0.91-1.0). The proportion of agreement for abnormal tests was 0.79 (95% CI: 0.57-1.0). The OKP test may be useful in glaucoma case-finding in general practice. However, the proportion of false positive tests seems to be too high for OKP to be used alone in mass screening. PMID- 8729018 TI - Is the incidence of constant esotropia in childhood reducing? AB - Episodes of strabismus surgery in the under-14 year age group in West Berkshire have reduced by 42%, from 22.7 to 13.2 per 10,000 population, between 1968 and 1985. Clinical audit of patterns of referral shows that the incidence of constant esotropia has reduced by 55%, from 28.3 to 12.8 per 10,000 population, between 1971 and 1991. PMID- 8729019 TI - Clinical management of age-related macular degeneration: room for improvement. AB - Age-related macular degeneration (AMD) remains the single most common cause of blind registration amongst the elderly population in the Western world. The majority of patients with AMD have untreatable disease, whilst the minority who do have disease amenable to laser photocoagulation have a better visual prognosis the earlier treatment is performed. This study was designed to assess the clinical management of AMD in the United Kingdom from the patients' perspective, so locating areas for improvement. One hundred and fifty-eight completed postal questionnaires of 200 originally distributed randomly to members of a self-help group, the Macular Disease Society, are reported. One hundred and five ophthalmic units were attended, nearly half recently (i.e. first attendance by patient within the past 4 years). Only 6% of patients received any written information on AMD. Satisfaction with Low Visual Aid (LVA) clinic appointments and LVA use was high, but patients' opinions on the value of regular hospital attendance was conflicting. A structured management plan in which education plays an integral role provides the opportunity for optimum care for AMD patients with both treatable and untreatable disease. PMID- 8729020 TI - Should second eye cataract surgery be rationed? AB - To conserve limited resources, healthcare purchasers are questioning whether they should ration second eye cataract surgery after successful unilateral extraction. The effect this would have on overall cataract workload is unknown. The clinical indications for performing second eye cataract extraction were recorded from the medical notes of a random group of patients undergoing second eye surgery over a 1 year period. Twenty-six per cent of all cataract operations were on second eyes and, of these, 21.5% of operations were on patients with coexistent ocular pathology, requiring clear optical media for disease monitoring, and 18.4% were on patients who had failed to achieve a satisfactory result after unilateral cataract surgery (visual acuity 6/18 or worse). Another 43.6% had severe binocular visual disability due to their remaining cataract. Only 4.4% of all cataract operations performed during the study period were on second eyes of patients with no other ocular pathology who had had successful unilateral surgery and had only mild symptoms from their remaining cataract (visual acuity 6/12 or better in second eye). Therefore, the overall savings made by rationing second eye cataract surgery to those patients who have only mild visual disability would be slight. If the proportion of cataract operations performed as daycases under local anaesthetic were increased, sufficient resources would be released to enable all patients to achieve maximum binocular visual rehabilitation. PMID- 8729021 TI - Sport-related eye trauma: a survey of the presentation of eye injuries to a casualty clinic and the use of protective eye-wear. AB - The study investigated the workload offered by sport-related trauma to a busy casualty clinic and was carried out in two parts. The first part concerned compiling information about sport-related injuries presenting in 1 year to the outpatient eye clinic at Bradford Royal Infirmary. The second part attempted to investigate the awareness and the use of eye-protectors for squash by the student community of Bradford University, using a questionnaire. The sport-related injuries presenting to the casualty department were classified according to the sport involved. Analysis of the data shows that racquet sports, namely tennis, badminton and squash, jointly formed the highest percentage of presentations. In the case of squash and tennis injuries, follow-up treatment and/or admission to hospital was required in 100% of cases. These injuries could be reduced by wearing adequate eye-protectors. The questionnaire survey showed that only 9% of the squash players wore eye protection whilst playing, of whom 3.37% of the total had suffered lacerating injuries previously. Comparison with previous studies indicates that the pattern of eye injuries presenting to casualty departments, and the awareness of the general public concerning the use of eye-protectors, has not altered in recent years. Unless steps are taken, in terms either of mandatory use of eye-protectors and/or of educating the general public on the possible devastating effects of injury, these preventable injuries will continue to burden already overworked hospital casualty departments, draining both resources and expertise. PMID- 8729022 TI - Aetiology and prevalence of eye injuries within the United Kingdom fire service. AB - A survey was conducted on a sample of 1552 firefighting personnel to determine the incidence of eye injuries within the United Kingdom fire service. In total, 3.5% (n = 22/637) of firefighters per year will sustain an eye injury requiring 'medical attention'. This represents one ocular injury per 50,000 working hours. In 80% of cases eye protection was not being worn at the time of injury. Analysis of 75 accident report forms indicates that the vast majority (69/75) of eye injuries were sustained whilst on operational duty. These statistics are comparable to other professions which are considered to have an unacceptably high risk of injury and in which eye protection has therefore been actively enforced. It would follow that similar steps should be taken within the fire service to ensure firefighters wear eye protection on operational duty. This should apply even if there is no immediate recognition of a perceived danger. A firefighter presenting to an optometrist or medical practitioner, with or without an eye injury, should be counselled as to the advantages of eye protection. PMID- 8729023 TI - The assessment of clinical investigations: the Greenwich Grading System and its application to electrodiagnostic testing in ophthalmology. AB - The aim of the study was to describe the Greenwich Grading System, a simple method developed for assessing the value of a clinical investigation, and to investigate its application to electrodiagnostic testing (EDT) in ophthalmology patients. This was done by means of a retrospective case analysis carried out on 95 unselected, consecutive patients referred from a district ophthalmology department for electrophysiological investigation at a regional neurosciences centre. The classification was developed to quantify the effect of EDT on the diagnosis, investigation and treatment in each case. From this, an overall value of EDT in management could be assessed for different groups of patients. It was found that the classification was easy to apply and analyse. EDT was valuable in 80% of patients referred, and was classified as essential or important in 53 (56%) patients. In no patient did EDT adversely affect the clinical outcome. The diagnosis was changed in 7 (7%) patients, and made in 5 (5%). Diagnostic certainty was increased in a further 62 (65%) patients. Other investigations were avoided by performing electrophysiology in 28 (29%) cases. Treatment was initiated in 6 (6%) cases and altered in 8 (8%). It is concluded that the Greenwich Grading System, as applied to EDT in ophthalmology, demonstrated the value of electrophysiological investigation in ophthalmology patients and identified the patient groups which benefit most from testing. PMID- 8729024 TI - Rationalising ophthalmic admissions. AB - A new ophthalmic admission document has been devised which integrates existing medical and nursing input promoting a more consistent and concise assessment of the patient. PMID- 8729025 TI - General practice referral letters to a regional ophthalmic accident and emergency department. AB - A significant proportion of patients seen in ophthalmic Accident and Emergency departments are referred by other medical practitioners. Two hundred and twenty six referral letters from general practitioners to the Accident and Emergency Department of a large regional ophthalmology centre were examined. Information that was poorly recorded in the referral letters included social and personal background details, examination findings, management plans and investigation results. Analysis revealed that 50-70% of referrals did not constitute accidents or urgent conditions and could have been more properly assessed within usual secondary referral outpatient clinics. Ophthalmic Accident and Emergency is therefore providing a service for general practitioner referrals in excess of its defined function. The impact on purchaser/provider arrangements if appropriate redirection to outpatients were to occur would be significant. PMID- 8729026 TI - Small-angle X-ray scattering of kappa- and iota-carrageenan in aqueous and in salt solutions. AB - Aqueous and saline (0.03-0.08 M NaCl) solutions of the Na(+)-salt of kappa- and iota-carrageenan (3-17 mg/ml) have been investigated using small-angle X-ray scattering (SAXS). Investigation of the SAXS curves normalized for concentration allows to detect the transition from the dilute to the semidilute regime. Some of the experimental curves exhibit a clear maximum. In the case of kappa carrageenan, the position of this maximum is proportional to the cube root of polysaccharide concentration. The fluctuation theory of Borue and Erukhimovich (BE theory), which predicts a maximum in the scattering curves for low ionic strength, gives a good fit for all our experiments. An increase of the polymer concentration or the ionic strength causes the appearance of two subsystems in solution exhibiting a different characteristic screening scale of Coulombic interaction for ideal Gaussian chains (and two different segment lengths). The condition for an increase of the relative contribution of the subsystem with the larger segment length correlates with the one for the disorder-order transition of carrageenan. In the most concentrated (17.1 mg/ml) solution of Na(+)-iota carrageenan, the increase of NaCl molarity above 0.06 M gives rise to additional scattered intensity at the smallest angles, indicating molecular association. PMID- 8729027 TI - Studies on the thermokinetic characterisation of actin polymerization and the effect of cisplatin. AB - The microcalorimetic measurement for studying the thermokinetic of actin polymerization in vitro was employed for the first time. The thermodynamic parameters of actin polymerization at 310.15 K are reported as delta Hm = 49.26 kJ/mol, delta Gm = 25.62 kJ/mol and delta Sm = 241.54 J/K.mol. The thermogram shows that polymerization of actin is a complicate process in which multistep reactions occur corresponding to exothermic and endothermic effects. Cisplatin, at lower concentration, affects the polymerization of G-actin resulting in a decrease of delta Hm; and, at higher concentration, induce the crosslinking and depolymerization of F-actin in the equilibrium system of G/F. The experimental results show that as the concentration of cisplatin increases, the sign of delta Hm changes from positive to negative, i.e. the magnitude and sign of delta Hm of actin polymerization depends on the concentration of cisplatin. PMID- 8729028 TI - The folding intermediate of reversibly denatured human prostatic acid phosphatase. AB - Human prostatic acid phosphatase (hPAP) [EC 3.1.3.2], a homodimer of ca. 50 kDa subunit molecular weight, shows reversible denaturation in 6 M urea at pH 2.5. Rapid dilution of the denatured enzyme allowed partial renaturation of hPAP, as measured by enzyme activity, to a level which depended on the composition of the dilution solution employed and time of the reaction. The renaturation reaction of hPAP was examined using spectral analysis (circular dichroism and fluorometry), fast size-exclusion chromatography and proteolysis by trypsin. The observed results are in agreement with the concentration-dependent kinetics of hPAP reactivation, assuming that the reconstitution of the active enzyme requires the association of subunits in dimeric form. Moreover, it suggests formation of an inactive intermediate during refolding of the denatured PAP. A mechanism of renaturation of the active enzyme from denatured PAP is proposed. PMID- 8729029 TI - Effect of electrostatic interaction on phase separation behaviour of chitin crystallite suspensions. AB - Optical and electron microscopy were used to observe the dynamics of the phase separation in aqueous chitin suspensions prepared by HCl hydrolysis of crab chitin. Freeze-fracture transmission electron microscopy reveals that chitin crystallites are partially aggregated in the suspension and have an average length of 200 nm and an average width of 8 nm. They exhibit a positive surface charge of approximately 0.5 e/nm2 when fully protonated. The liquid crystal forming aqueous suspensions of such crystallites are investigated through phase diagrams and Zeta potential measurements for different ionic strength. Exposure of the suspension to a low concentration of univalent electrolyte has a negligible effect on phase separation because of the contribution of the charged crystallites themselves to the ionic strength. The thickness of the effective repulsive layer is estimated both from the phase diagrams according to Onsager's theory and from the computed interaction energy derived from the Poisson Boltzmann equation using the experimental Zeta potential as surface potential. When the contribution of crystallites to the ionic strength is taken into account and an hypothetical linear charge density close to the Manning limit is assumed, there is good agreement with the Debye length. PMID- 8729031 TI - Single crystals of inulin. AB - Lamellar crystals of inulin were grown by crystallizing sharp fractions of low molecular weight inulin from dilute aqueous ethanol solutions. The crystals were analyzed using three-dimensional electron diffraction and X-ray powder diagrams. Two crystalline polymorphs were observed, depending on the hydration conditions: a hydrated form which indexed on an orthorhombic unit cell, with space group P2(1)2(1)2(1) and with cell dimensions of a = 1.670 nm, b = 0.980 nm and c (chain axis) = 1.47 nm, together with a pseudo-hexagonal semi-hydrated form with unit cell parameters a = 1.670 nm, b = 0.965 nm and c (chain axis) = 1.44 nm. These parameters, together with the density data, indicate that inulin crystallizes along a pseudo-hexagonal six-fold symmetry with an advance per monomer of 0.24 nm. The difference between the hydrated and the semi-hydrated unit cells does not seem to correspond to any change in the conformation of inulin, but rather to a variation in water content. PMID- 8729030 TI - Distribution of solvent and ligand molecules around aromatic side chains in proteins and its implication on carbonic anhydrase catalytic mechanism. AB - Interactions between aromatic phenylalanine, tyrosine and tryptophan residues and water as well as ligand molecules were analyzed by a computer search of a large number of high resolution structures in the protein data bank. Water molecules and oxygen ligands have a preference for the edges of the aromatic residues, but the distribution of carbon ligands around the aromatic rings is different. The more hydrophilic tyrosine, followed by tryptophan, has the highest frequency of water contacts. However the situation is reversed for ligand interactions where phenylalanine is the most active of the three aromatic residues. The results indicate that hydrogen bonding by water molecules to aromatic pi-electron does not occur in protein structures. The role of the conserved Trp 209 in the catalytic mechanism of carbonic anhydrase is discussed in consideration of the results from the data base search. Normally not considered as a catalytic residue, this tryptophan is proposed to participate in the physiologically important enzymatic interconversion of carbon dioxide and bicarbonate by stabilizing substrate coordination through van der Waals' interactions. PMID- 8729032 TI - Electrostatic parameters of the theoretical quaternary structure of bovine alpha crystallin. AB - By changing the ionic strength, the effects of charge modification on the electrostatic properties of our predicted "open' micellar quaternary structure composed of bovine alpha A subunits were determined. The electrostatic potential values (phi) at 6 arbitrary points surrounding the protein and at all atomic sites of the protein were calculated using the non-linear Poisson-Boltzmann equation. The effective charge (q) of our predicted aggregate ranged from 16 at 0.0022 M to 45 at 0.1472 M ionic strengths. The variation of potential (phi), as well as charge, is a hyperbolic function of ionic strength (R2, 0.901). Plotting the inverse charge (l/q) against inverse ionic strength (1/l) it is possible to calculate maximum charge (q(max)) (approximately 48) at saturation. This value is close to previously reported experimental (50 +/- 5), and our theoretical charge (45), values at physiological ionic strength (0.145 M). These data indicate that maximal repulsion among the alpha-crystallin aggregates occurs at or near physiological ionic strength. Also, half-maximal charge (q(max)/2) at 0.003-0.004 M indicates a transition state at very low ionic strength. The calculated volume available for the mobile solvent in our quaternary structure is approximately 70%. These data are in good agreement with experimental values for bovine alpha crystallin in solution reported by Xia et al. (Biophys. J., 1994; 66: 861-872). This agreement provides support for our predicted models of alpha-crystallin and a level of confidence in the reliability of the theoretical calculations. Since an ionic gradient exists between the lens cortical and nuclear regions, the modification of charge on alpha-crystallin by varying ionic strength could contribute to the function of alpha-crystallin as a modulator of lens supermolecular order during fiber cell maturation. PMID- 8729033 TI - Conformations and flexibility of native and re-natured xanthan in aqueous solutions. AB - The conformation and flexibility of sonicated 'native' and 're-natured' xanthan have been investigated by size exclusion chromatography (SEC) with coupled multi angle light scattering and viscosity detectors. 'Native' xanthan (NX) refers to xanthan dissolved in moderate ionic strength aqueous solution, which has not been exposed either to high temperature or very low ionic strength, and 're-natured' xanthan (RX) here refers to xanthan which has been heated above the conformational melting temperature and then recooled. The mass distributions of the NX and RX are virtually identical, implying that the RX does not involve aggregates of, or disassociated fragments of, NX. The flexibilities and conformations between NX and RX, however, are strikingly different; RX is far stiffer than NX, the persistence lengths being roughly 1000 A and 300 A, respectively, and the mass per unit length M/L of the RX is roughly double that of NX. With estimated M/L of 200 Da/A and 98 Da/A, respectively, the results strengthen the notion that RX is double stranded, whereas as NX appears single stranded. The nature and mechanism of formation of the double-stranded form is still unclear, and a few speculative scenarios are suggested. Finally, preliminary results on the kinetics of xanthan self-association in HCI are presented which illustrate the complexity of such processes in xanthan. PMID- 8729034 TI - On the structure of aggregated kappa-carrageenan helices. A study by cryo-TEM, optical rotation and viscometry. AB - Cryo-transmission electron microscopy (cryo-TEM), viscometry and optical rotation have been used to study the microstructure of dilute solutions of helical kappa carrageenan under varied electrolyte composition (0.1 M of NaI, CsI and mixtures of the two salts). Microfibers (300-400 nm in length) were observed above a critical cesium content, while below that no microstructure was seen in the micrographs. A drastic increase in the specific viscosity and an onset of thermal hysteresis in the optical rotation above the same critical cesium content provide clear evidence of aggregate formation. This leads to the conclusion that the observed microfibers are aggregates consisting of several helices. At high proportions of cesium, further aggregation of the microfibers was observed by cryo-TEM. PMID- 8729035 TI - Nitrosyl hemoglobins: EPR above 80 K. AB - The EPR spectra of nitrosyl hemoglobin and myoglobin in different conditions (native, denatured and lyophilized), as well as of hematin-NO were obtained in the temperature range of 80-280 K. There is a substantial and reversible decrease of the areas of the EPR spectra of all the hemoglobin samples above 150 K. The interpretation of the results implies the existence of two conformational states in thermal equilibrium, only one of which is EPR detectable. Thermodynamical parameters are determined for the hexa- and penta-coordinated cases. PMID- 8729036 TI - Evaluation of different absorbance ratios from infrared spectroscopy for analyzing the degree of deacetylation in chitin. AB - Various acetylated chitosan derivatives and mixtures of chitin and chitosan, covering the range of the degree of deacetylation (DDA) from 0-100% were analyzed by 1H-NMR spectroscopy and infrared spectroscopy. The use of the 1070 cm-1 or 1030 cm-1 absorption band as an internal standard in the determination of DDA from the absorbance of the amide I bands at 1655 cm-1 and 1630 cm-1 or the amide II band at 1560 cm-1 was studied. There is a good correlation between the results from IR spectroscopy and those from 1H-NMR spectroscopy. PMID- 8729037 TI - Histopathology of the liver in non-cirrhotic portal hypertension of unknown aetiology. AB - Non-cirrhotic, long-standing portal hypertension of unknown aetiology is being re evaluated histopathologically and clinically. In this study, we examined 107 livers with this condition (92 wedge biopsy and 15 autopsy specimens) from five institutions in Japan. These cases were histologically categorized into four groups: idiopathic portal hypertension (66 cases), nodular regenerative hyperplasia (14 cases), partial nodular transformation (two cases), and incomplete septal cirrhosis (25 cases). These four groups shared several histological features: dense portal fibrosis with portal venous obliteration and intralobular slender fibrosis. In addition, the histopathological features characteristic of one group were also found to a mild degree in other groups. The histopathological lesions preceding portal venous obliteration remain speculative. However, the portal venous obliteration may be responsible for the occurrence of sustained portal hypertension and several of the pathological changes in these livers. It seems likely that idiopathic portal hypertension, nodular regenerative hyperplasia, partial nodular transformation and incomplete septal cirrhosis comprise a family of non-cirrhotic, long-standing portal hypertension in Japan, and the histological differences between them may reflect chronological progression of a single disease. PMID- 8729038 TI - Accessory cells in physiological lymphoid tissue from the intestine: an immunohistochemical study. AB - We report a study of the organization of accessory cell populations, in normal mucosal lymphoid tissue from small intestine (8 cases), large intestine (6) and appendix (9) using a panel of monoclonal antibodies and polyclonal antisera in paraffin-embedded tissue. Two populations were identified in dome areas, one positive for acid cysteine proteinase inhibitor and HLA class II (WR18) only and the second positive for S-100 protein, CD68, and WR18 and negative for acid cysteine proteinase inhibitor and factor XIIIa. Superficial colonic mucosal and small intestinal villous tip macrophages stained positively with CD68 and WR18 only, while deeper cryptal and submucosal populations exhibited additional positivity for factor XIIIa, but both populations were negative for acid cysteine proteinase inhibitor and S-100 protein. Germinal centre macrophages were positive for CD68, WR18 and acid cysteine proteinase inhibitor and negative for factor XIIIa, and S-100 protein. T zone dendritic cells included a population which stained positively for S-100 protien, WR18 and were negative for factor XIIIa, CD68 and acid cysteine proteinase inhibitor, an immunophenotype typical of interdigitating dendritic reticulum cells. This distribution of phenotypically identifiable accessory cell subpopulations was apparent at all three sites examined. We suggest that the specialized subpopulations of dendritic cells staining for S-100 protein and for acid cysteine proteinase inhibitor which are restricted to the dome areas, may have a potential role in the transfer of antigen across the epithelium to the germinal centres, while factor XIIIa appears to identify a tissue macrophage population with a potential role in stromal modulation distant from direct antigen challenge. PMID- 8729039 TI - Accessory cells in Crohn's disease of the terminal ileum. AB - We present a study which describes the immunophenotype and distribution of accessory cells in 13 resections of terminal ileum from patients with Crohn's disease. A panel of antibodies working in paraffin-embedded tissue was employed and these included PGM1 (CD68), S-100 protein, WR18 (HLA class II), factor XIIIa and acid cysteine proteinase inhibitor. This study revealed a heterogeneity of accessory cell populations which was profoundly influenced by local inflammatory and repair mechanisms. Both acid cysteine proteinase activity and S-100 protein positive cells are identified in more actively inflamed areas. The acid cysteine proteinase activity positive dendritic cell population was particularly numerous in ulcer bases. S-100 protein positive dendritic cells had a more limited distribution in close proximity to the epithelium in inflamed but otherwise intact mucosa adjacent to the areas of ulceration. PGM1 revealed normal distribution of macrophages within histologically uninvolved areas and, in addition, also stained granulomas and large numbers of dendritic cells in the inflamed, ulcerated and scarred areas. Factor XIIIa positive dendritic cells were especially numerous in areas of active scarring where they co-localized with PGM1 positive cells. They were largely absent from the more superficial ulcerated areas. HLA class II was strongly expressed on mononuclear inflammatory and dendritic cells. The strength of epithelial staining for HLA class II reflected the intensity of adjacent inflammation, except on ulcer-associated epithelium which consistently showed up-regulation independent of the severity of the inflammatory process. This study shows that localized alterations in the accessory cell distribution in Crohn's disease correlate with different states in the evolution of the inflammatory and repair process of the disease. The more acute lesions are associated with recruitment of acid cysteine proteinase activity and S-100 protein positive dendritic cells while factor XIIIa stained dentritic cells are especially numerous in areas of scarring. PMID- 8729040 TI - EGF-receptors in human normal and pathological thyroid tissue. AB - Expression of the epidermal growth factor receptor (EGFR) was studied in cryosections from human thyroid tissues. Normal tissue (4 cases), nodular goitre (12), toxic goitre (9), adenoma (9), follicular carcinoma (1), papillary carcinoma (7) and poorly differentiated carcinoma (1) were used for immunohistochemistry. Northern blot analysis was performed in two nodular goitres, three adenomas, two papillary carcinomas, one follicular carcinoma and the adjacent normal tissue in five cases as well as in two cell lines from anaplastic carcinomas. Epidermal growth factor receptor immunoreactivity was detected in all tissues examined. The amount of EGFR mRNA did not differ between normal and abnormal tissues. However, the EGFR staining was weaker in normal thyroid tissue compared to the adjacent neoplastic areas suggesting an upregulation at the posttranslational level in the latter. A strong staining was also seen in hyperfunctioning thyroid glands. The EGFR location was mainly basal or basolateral in all thyroid tissues with normal histology and in toxic diffuse goitre. Pericellular and sometimes cytoplasmatic staining was seen in neoplastic tissues. In nodular goitre the staining was both basal, lateral and apical and varied in intensity. Our data suggest that a non-polarized location of EGFR probably indicates a loss of the normal epithelial cell polarity and could be interpreted as an early sign of dedifferentiation. Furthermore, a role for the EGFR is proposed, not only in the development of thyroid neoplasias but also in goitre formation. PMID- 8729041 TI - Massive ovarian oedema. AB - Eighteen cases of massive ovarian oedema are presented. The age of patients averaged 26 years and 16 presented with an acute abdomen. Hormonal symptoms included virilism in three cases and one with precocious pseudopuberty. Ultrasonographic findings were variable and not diagnostically accurate. When performed, CA 125 levels were not raised. Seventy-two percent of cases occurred in the right ovary and none were bilateral. Torsion occurred in 14 cases. Salpingo-oophorectomy was performed in all cases. To elucidate its pathogenesis, be this either due to intermittent chronic torsion or to a proliferative phenomenon, immunohistochemistry for Ki-67 and PCNA proliferation antigens, alpha actin and oestrogen and progesterone receptors was performed. The Ki-67 proliferation index ranged between 0% and 3%, demonstrating the low proliferative status of stromal cells. The PCNA indices, however, were unusually high (60% and above). The divergence between these findings is explained by the fact that PCNA positivity may be related to nuclear reparation subsequent to ischaemia. Alpha actin was consistently positive in stromal cells, reflecting a myofibroblastic transformation of these cells. These findings together with the clinical evidence of torsion in the majority of cases, lead us to consider that ovarian oedema is a reactive, non-proliferative state of specific stromal cells, occurring as a response to torsion and subsequent ischaemia. The stromal cells have positive oestrogen progesterone receptors and may undergo stimulatory changes responsible for the hormonally related symptoms often found associated with massive ovarian oedema. PMID- 8729042 TI - An immunocytochemical assessment of 19 cases of cutaneous angiosarcoma. AB - Four endothelial cell markers, two selective cytokeratin markers and a monoclonal smooth muscle antibody (SMA) were employed in the assessment of 19 cases of cutaneous angiosarcoma classified according to their degree of tumour differentiation. No labelling was seen for SMA or with cytokeratin markers MNF116 and CBL170. Expression of factor VIII-related antigen was seen in two tumours and positivity for CD34 (QBend 10 antibody) was found in four tumours. By contrast the pan-endothelial cell marker Ulex europeaus agglutinin 1 (UEA-1) and the CD31 marker JC70A labelled all cases of cutaneous angiosarcoma with the exception of one poorly differentiated tumour. These data confirm the endothelial cell origin of angiosarcoma, they demonstrate that CD31 and UEA1 are reliable markers in routinely processed tissue, and they suggest a lymphatic derivation for the tumour. This finding is in marked contrast to Kaposi's sarcoma where CD34 is the most reliable marker. PMID- 8729043 TI - Breast cancer: two micrometastatic variants in the axilla that differ in prognosis. AB - Measurement of the area of the tumour deposits present in routine sections from the axillary nodes from a series of 1069 breast cancer patients showed that 138 cases had a single micrometastasis (0.2 cm2 or less), while in 29 a similar load was spread over two or more nodes. These 167 cases represent 15% of the patients in the series. Twentyfive of them had died of breast cancer within a mean follow up of 6 years. They had smaller micrometastases than those surviving (P < 0.0025). Histological examination in the 138 with single micrometastases showed that two variants were present. In one, tumour growth was confined to the capsular lymphatics and/or the subcapsular sinus. In the other, tumour growth was present in the nodal lymphoid tissue, and, on occasion, at the other sites as well. Those with growth in the lymphoid tissue had a better prognosis than those without (P < 0.0035). Prognosis in the former was comparable to that in the node negative cases, while in those lacking such growth it was similar to that in the node-positive. The presence of these two variants could explain divergent reports in the literature on prognosis in cases with micrometastases. While the mechanisms behind this apparent paradox remain speculative, the observation can be of diagnostic interest in routine surgical pathology. PMID- 8729044 TI - Immunohistochemical study of c-erbB-2 expression in carcinoma ex-pleomorphic adenoma. AB - A series of 19 cases of carcinoma ex-pleomorphic adenoma was studied for the immuno-expression of c-erbB-2 oncoprotein. Twelve tumours showed a malignant component with only one histological type; in the remaining seven there was co existence of areas of various carcinoma types, adenocarcinoma NOS being the most frequent. Membranous c-erbB-2 reactivity was found in 21.1% of the cases, all corresponding to high-grade adenocarcinomatous areas. The low-grade carcinoma types that formed the malignant mixed tumours components were negative. Benign pleomorphic adenoma areas, either adjacent or intermingled with carcinomatous areas, were also consistently negative, proving that c-erbB-2 accumulation is associated with the acquisition of the malignant phenotype. The finding of a preferential association between c-erbB-2 overexpression and high-grade malignant mixed tumour may indicate prognostic implications for the oncogene protein and may also be indicative of its specific relationship with the putative pathway of malignant transformation in pleomorphic adenomas. PMID- 8729045 TI - Alkaline encrusted cystitis associated with malakoplakia. AB - Alkaline encrusted cystitis is a rare inflammatory condition of the bladder which has been reported sporadically over the past 80 years. It is caused by infection with urea splitting organisms leading to the deposition of inorganic salts on to the surface of the bladder. We present three cases of alkaline encrusted cystitis. In two cases the encrusted area was associated with foci of malakoplakia. The third case occurred in a patient who had received chemotherapy for acute lymphoblastic leukaemia. To our knowledge, these are the first cases of malakoplakia associated with alkaline encrusted cystitis. These two conditions have a number of clinical and aetiological similarities, and may have more in common than has been previously thought. PMID- 8729046 TI - EBV latent membrane protein (LMP-1) and bcl-2 protein expression in Reed Sternberg-like cells in post-transplant lymphoproliferative disorders. AB - An inconsistent association exists between EBV-LMP-1 and bcl-2 protein expression in Reed-Sternberg cells seen in Hodgkin's disease. In fact, many studies have concluded that there is no correlation between EBV-LMP and bcl-2 expression in Hodgkin's disease. We undertook an analysis of post-transplant lymphoproliferative disorders to explore the relationship between EBV-LMP and bcl 2 in Reed-Sternberg-like cells found in this condition, given the strong association between this disorder and EBV. Reed-Sternberg-like cells were found histologically in 11 of 28 cases of renal, heart and heart-lung post-transplant lymphoproliferative disorders. Formalin-fixed, paraffinembedded sections were stained with monoclonal antibodies to EBV-LMP-1 and bcl-2 proteins. Reed Sternberg-like cells in all 11 cases co-expressed EBV-LMP and bcl-2. A similar relationship was noted with large, mononuclear cells and occasional small lymphoid cells. The staining pattern seen with both antibodies was of similar intensity and both displayed cytoplasmic Golgi accentuation. In the setting of post-transplant lymphoproliferative disorders. Reed-Sternberg-like cells exhibit strong co-expression of EBV-LMP-1 and bcl-2 proteins, supporting a positive correlation between them. This is in contrast to the findings in Hodgkin's disease. The reason for this discrepancy may be due to the iatrogenic immunosuppression and resultant severe EBV infection, together with other cellular events. PMID- 8729047 TI - Solitary fibrous tumour of the major salivary glands. AB - We report four cases of solitary fibrous tumours which involved the parotid (two cases), submandibular, and sublingual glands of two men and two women ranging in age from 46 to 81, mean 66 years. The tumours presented as painless, firm masses which involved the substance of the salivary gland in each case. All have had an uneventful clinical course after local excision. Awareness of this entity is important to avoid confusion with haemangiopericytoma. PMID- 8729048 TI - Granulomas and hepatitis C. AB - Hepatitis C (HCV) is associated with a number of characteristic histological features. A recent paper has identified an increased frequency of granulomas in resection specimens from cases of HCV. We have carried out a retrospective study of 155 cases of HCV to assess the frequency of granulomas in biopsy specimens. We had two control groups: 151 cases of hepatitis B (HBV) and 129 cases of alcohol induced liver disease. Granulomas were found in 14 cases of HCV (10%), three cases of HBV (2%) and three cases of alcohol induced liver disease (2%). Granulomas were significantly commoner in cases of HCV than in the other two groups. Of the 14 cases of HCV, the granulomas could be ascribed to another cause in seven cases. When the analysis was carried out, excluding those granulomas which could be ascribed to another cause, they were still significantly commoner in cases of HCV. We conclude that granulomas are more frequent in HCV but that in half of cases in which they are found another cause can be identified. This means that if granulomas are seen in association with hepatitis C another aetiology should be sought before ascribing them to HCV. PMID- 8729049 TI - Limited value of the NKI/C3-antibody for the differential diagnosis of Paget's disease of the nipple and intra-epidermal malignant melanoma. PMID- 8729050 TI - Orthotopic liver transplantation for type 2 hepatic infantile haemangioendothelioma. PMID- 8729051 TI - Non-progressive malakoplakia in a live donor renal allograft. PMID- 8729052 TI - Epithelioid cell variant of renal angiomyolipoma. PMID- 8729053 TI - Kaposiform haemangioendothelioma associated with Kasabach-Merritt syndrome. PMID- 8729054 TI - Unusual cystadenomatous hamartoma of the lung. PMID- 8729055 TI - Altered interactions among thin filament proteins modulate cardiac function. AB - The transition of cardiac muscle myofilament activity from the diastolic to the systolic state is a complex allosteric/co-operative process, rich in potential control mechanisms. The rate and intensity of the transition is modulated by the mechanical state of the myofilaments, by covalent and non-covalent mechanisms, and by the isoform population of myofilament proteins. Moreover, the process is altered in pathological states and subject to modification by pharmacological agents with potential use as inotropic drugs. We present here a current perception of the process, with focus on molecular interactions of the thin filament components, especially troponin I. Our discussion is couched in terms of what we believe to be pressing questions in the current state of knowledge of this system. These questions are as follows: what is the topology of the thin filament and how do thin filament proteins regulate the activation of cross bridge cycling? What is the relative role of protein phosphorylation of thin filament proteins in the regulation of the cardiac activity and dynamics? What is the relative role of feedback effects of cross-bridge binding on thin filament activity? Answers to these questions have taken on new significance, with the identification of familial hypertrophic cardiomyopathy as a "sarcomeric" disease related to missense mutations in myosin, troponin T. and tropomyosin. As discussed, new and exciting developments in this area are bringing us closer to the answers to these questions. PMID- 8729056 TI - A brief period of retrograde hyperthermic perfusion enhances myocardial protection from global ischemia: association with accumulation of Hsp 70 mRNA and protein. AB - The induction of heat shock proteins in the myocardium has been suggested as a possible intervention to allow for enhanced cardioprotection. We examined the cardioprotective effects of Hsp 70 induction in a clinically relevant model, in which a brief period of retrograde hyperthermic perfusion (42 degrees C) was applied for 15 min, only 5 min prior to global ischemia and reperfusion in the isolated perfused rat heart. Our results indicate that in retrograde hyperthermic perfused hearts (n = 17) there was enhanced dP/dT, end diastolic pressure, and peak developed pressure during normothermic reperfusion following 15 min of global ischemia when compared to control hearts perfused at 37 degrees C (n = 18). Northern analysis indicated Hsp 70 mRNA, in retrograde hyperthermic perfused hearts, was increased 9.0 +/- 0./70-fold (P < 0.001) by 30 min and 9.1 +/- 0.45 fold (P < 0.001) by 60 min of normothermic reperfusion. Western analysis revealed that the Hsp, heat inducible 72 kD protein was increased 1.74 +/- 0.35-fold (P < 0.001) by 30 min and 1.79 +/- 0.31-fold at 60 min of normothermic reperfusion when compared to no ischemia hearts. Our results demonstrate that the use of 15 min of retrograde hyperthermic perfusion, only 5 min prior to global ischemia and reperfusion, provide for enhanced myocardial functional recovery. Enhanced myocardial functional recovery was associated with the accumulation of Hsp 70 mRNA and the Hsp 72 kD protein. PMID- 8729057 TI - The effects of endothelin-1 on collagen type I and type III synthesis in cultured porcine coronary artery vascular smooth muscle cells. AB - Restenosis is the single most important factor limiting a favorable long-term outcome following mechanical revascularization. The vascular endothelium, through the release of key regulatory compounds, may regulate vascular structure by exerting fundamental control over collagen synthesis following injury to the vessel wall. We tested the hypothesis that endothelin (ET-1), an endothelium derived peptide previously shown to be increased in pathological states, differentially stimulates porcine coronary vascular smooth muscle cell collagen types I and III synthesis. Monocultures of porcine coronary vascular smooth muscle were exposed to varying concentrations of endothelin over a 24-96-h time period. The medium was assayed for soluble collagen types I and III using a sensitive and specific ELISA method. Experiments were also done with the ET-1 antagonists PD 145065 and BQ123. Cell counts and viability were serially monitored. Experiments were also conducted with angiotensin II (A-II). A-II and ET-1 stimulated cell proliferation. ET-1 maximally stimulated collagen type I synthesis at 48 h at an optimal concentration of 10(-8) M, with no significant stimulation of collagen type III synthesis. The ETA specific antagonist BQ123 significantly inhibited the stimulatory effects of ET-1. A-II also stimulated collagen type I synthesis above basal levels, but was less efficacious than endothelin (95 +/- 5%, A-II, v 189 +/- 14% ET-1). In contrast to ET-1, A-II stimulated collagen type III synthesis (31 +/- 6% above basal, compared to -4 +/- 5% for ET-1). Results are also reported using smooth muscle cells from porcine aorta. The data demonstrate that ET-1 and A-II stimulate collagen synthesis by coronary artery vascular smooth muscle, and that they exert a differential effect over the two types of collagen that are present in the intima following balloon injury. Thus, the over expression of key regulatory compounds by endothelium following balloon injury could enhance collagen deposition and, consequently, play an integral role in intimal hyperplasia and restenosis. PMID- 8729058 TI - Functional coupling between sarcoplasmic reticulum and Na/Ca exchange in single myocytes of guinea-pig and rat heart. AB - It has been proposed that the cardiac Na/Ca exchanger is primed by high Ca2+ concentrations derived from the sarcoplasmic reticulum (SR) in a recently identified subsarcolemmal, "Na/Ca exchange-dependent Ca2+ compartment". We tested this hypothesis by investigating the effect on Na/Ca exchange of interventions affecting Ca2+ flux through SR. Experiments were performed in single, isolated myocytes of guinea-pig and rat hearts loaded with Indo 1-AM and free Ca2+ concentration was assessed by measuring the ratio of fluorescence at 405 and 495 nm wavelength. In guinea-pig 1.0 microM ryanodine (Ry), expected to increase Ca2+ flux through the SR, decreased the amplitude of electrically stimulated Ca2+ transients by 35% and inhibited their initial, rapid phase. Responses of these cells to brief superfusions with 15 mM caffeine were inhibited which suggests that 1.0 microM Ry depleted the SR Ca2+ stores. Diastolic Ca2+ concentration was slightly increased, but it dropped below control during prolonged rest. Decrease of the amplitude of the transients in these ryanodine-treated cells were reversed by 2 x 10(-7) M thapsigargin (TG), an inhibitor of the SR Ca(2+)-ATPase, by 1.0 mM Ry, a blocker of the SR Ca2+ release channels and by low Na+ (50.0 mM) superfusion. This suggests that the decreased transients in 1.0 microM Ry result from uptake of Ca2+ by the SR and its rapid release to the subsarcolemmal (cleft) space where a fraction is diverted out of the cell via Na/Ca exchange before it can diffuse to the myofilaments. Removal of the SR from the pathway (addition of TG on 1.0 mM Ry) or reversal of Na/Ca exchange diverts more transsarcolemmal Ca2+ influx to the myofilaments and increases the Ca2+ transient. Decrease of the resting Ca2+ concentration was blocked by 2 x 10(-7) M TG, 1.0 mM Ry, and by 5.0 mM Ni2+, a blocker of Na/Ca exchange. This result suggests that the effect of 1.0 microM Ry on resting Ca2+ concentration also resulted from increase of flux of Ca2+ through the SR and out of the cell by Na/Ca exchange. In rat 1.0 microM Ry decreased amplitude of the transients by approximately 75% but did not affect their kinetics. TG and 1.0 mM Ry decreased the rate of rise of the transients and greatly delayed their decay. This result suggests that normal kinetics of the transients in the cells treated with 1.0 microM Ry depended on the preserved Ca2+ flux through the SR. As SR was not able to retain Ca2+ in these cells, decay of the transients must have depended on stimulated Na/Ca exchange. The results in guinea-pig and rat taken together are compatible with the proposal that Ca2+ released from the SR interacts with the cell's Na/Ca exchanger most probably within the newly defined subsarcolemmal "Na/Ca exchange-dependent Ca2+ compartment". PMID- 8729059 TI - Modulation of endothelin-1 effects on rat hearts and cardiomyocytes by nitric oxide and 8-bromo cyclic GMP. AB - Endothelin-1 (ET-1) has been demonstrated to produce numerous cardiac effects and increased production of the peptide has been shown in cardiac disease states. Although the cardiac effects of ET-1 have been examined extensively on its own, few studies have reported potential cross-talk between ET-1 with other endothelium-derived factors. We examined whether nitric oxide (NO) can modulate the effects of ET-1 on isolated rat hearts or ventricular myocytes. At 0.05 nM, ET-1 produced no effects on either systolic or diastolic function although a two fold increase in left ventricular end-diastolic pressure (LVEDP) was observed in hearts pretreated with 10 microM of the NO synthase inhibitor L-NAME. Higher concentrations of ET-1 (0.5 and 5 nM) produced a direct elevation in LVEDP which was enhanced by L-NAME and totally blocked by the NO donor S nitrosoacetylpenicillamine (SNAP, 10 microM) although responses to 5 nM ET-1 were highly variable with no significant differences between treatment groups. SNAP totally prevented ventricular fibrillation produced by either 0.05 or 0.5 nM ET-1 whereas the pro-fibrillatory actions of 5 nM ET-1 were unaffected. In cardiac myocytes, SNAP significantly attenuated the elevation in intracellular Ca2+ produced by ET-1 (5 nM). The positive inotropic actions of ET-1 on either hearts or myocytes were unaffected by any treatment. The protective effect of SNAP against ET-1 in both isolated hearts (reduction in LVEDP and incidence of fibrillation) as well as ventricular myocytes (attenuation of the elevation in intracellular Ca2+) was mimicked by 8-bromo-cyclic GMP (50 microM). Our study suggests that NO protects against the cardiotoxic effects of ET-1, possibly via inhibition of intracellular Ca2+ elevations, a property shared by cGMP, the likely mediator of the biological effects of NO. PMID- 8729060 TI - Early action potential shortening in hypoxic hearts: role of chloride current(s) mediated by catecholamine release. AB - We tested the hypothesis that the early action potential shortening induced by hypoxia in perfused hearts is attributable to chloride currents activated or modulated by endogenous catecholamine release. Rabbit hearts perfused at 33 degrees C and paced at 2.5-2.8 Hz were used for membrane potential recordings with microelectrodes. Catecholamine depletion was induced with reserpine treatment. The effects of nadolol (10 microM), the stilbenedisulfonic acid derivatives DIDS (10 microM) and SITS (1 mM), and diphenylamine-2 carboxylate (DPC, 100 microM) on action potential characteristics were determined at different times during hypoxia. The effect of chloride transport blockers on the outward currents induced by 200 nM carbonyl cyanide (CCCP) or by 1 microM isoproterenol in isolated cells was also tested. In control hearts, action potential duration (APD) at 25 and 95% repolarization decreased by 50 +/- 9% and 32 +/- 7% respectively after 5 min of hypoxia. This effect was fully antagonized by reserpine pretreatment, by respiratory acidosis, and by nadolol when present from the beginning of hypoxia. None of these agents affected action potential characteristics in normoxia and nadolol had no effect when added after 15 min of hypoxia. Lowering the chloride concentration to 17.5 mM reproduced the effects of nadolol and reserpine. DIDS and SITS lengthened APD in normoxia and attenuated the early APD shortening in hypoxia. DPC had no effect in normoxia but fully counteracted APD shortening produced by isoproterenol or early hypoxia. In isolated cells, DIDS did not affect the glibenclamide sensitive outward current induced by CCCP and DPC blocked the isoproterenol induced current. The data suggest that in whole hearts, chloride currents mediated by endogenous catecholamine release are involved in the early action potential shortening induced by hypoxia with preservation of glycolysis. PMID- 8729061 TI - Inability of the heat-shocked heart to adjust its pre-ischemic and post-ischemic performance to variable loading conditions. AB - The aim of the present study was to investigate whether the pre-ischemic and post ischemic hemodynamic function of the heat-shocked rat heart is affected by changes in afterload and extracellular calcium concentrations ([Ca2+]e). Experiments were performed on isolated, ejecting Lewis rat hearts 24h after in vivo heat shock (LewHS) or anesthesia alone (Lewc). In vitro hearts were subjected to 60 min normoxic perfusion, 45 min global ischemia, and 60 min of reperfusion. Pre-ischemic and post-ischemic left ventricular performance was evaluated at [Ca2+]e ranging between 0.65 and 3.0 mM at afterloads of 8.0 kPa and 16.0 kPa. At 8.0 kPa, pre-ischemic function was comparable in LewHS and Lewc at [Ca2+]e equal to or above 2.25 mM. At lower [Ca2+]e, i.e., 0.65 and 1.25 mM, cardiac output (CO) was significantly lower in LewHS than in Lewc hearts. At 16.0 kPa, significantly lower CO values were found in LewHS than Lewc hearts at all [Ca2+]e levels. During post-ischemic reperfusion under basal conditions (8.0 kPa; [Ca2+]e = 2.25 mM) a significantly better recovery was observed in LewHS than Lewc hearts, persisting at [Ca2+]e equal to 1.25 mM. However, either by lowering [Ca2+]e to 0.65 mM or increasing afterload to 16.0 kPa (at all [Ca2+]e), heat shock-associated improvement of post-ischemic performance disappeared. In conclusion, pre-ischemic left ventricular performance of the isolated heat shocked heart is depressed when it performs at low [Ca2+]e or against a relatively high afterload. The heat shock-mediated improvement of post-ischemic function is only present at relatively low afterload levels in combination with normal extracellular calcium concentrations. PMID- 8729062 TI - Transmembrane movement of lithium ions in isolated sheep heart Purkinje fibres. AB - The many routes by which Li can cross the cardiac sarcolemma were examined using Li-selective microelectrodes. In the presence of 70 mmol/l extracellular Li there was a rapid increase in intracellular Li activity (aiLi) equivalent to an influx of 0.45 mmol/l/min (or 2.1 pmol/cm2.s). Depolarisation (voltage-clamp or high [K]O) had no significant effect on the rate of aiLi increase, or recovery on removal of LiO. Following addition of 20 mmol/l [Li]O to the normal Tyrode, the rate of aiLi increase was stimulated by 49.7% by removal of extracellular Mg. However Li entry was inhibited by SITS (100 mumol/l) by 20.6%; tetrodotoxin (10(15)g/ml) 20.0%; caesium (2 mmol/l) 22.5%; verapamil (20 mumol/l) 14.3%; and manganese (1 mmol/l) 37.6%. With 5 mmol/l [Li]O, aiLi stabilized at 2.3 mmol/l, i.e. much lower than expected assuming passive distribution of Li (predicted level 61 mmol/l), implying active extrusion of Li from the cells. This stable level of Li was not significantly affected by short exposures to strophanthidin, bumetanide, or ethyl isopropylamiloride, but was decreased by increasing the [Ca]O or adding manganese. The aiLi was increased on removal of [Na]O or addition of phloretin (100 mumol/l) suggesting that NaO-Lii exchange helps to maintain a low aiLi. The removal of KO produced an increase in aiLi. This effect was inhibited by strophanthidin, suggesting Li can enter via the Na/K pump in K-free conditions. PMID- 8729063 TI - Cardiac injury by activated leukocytes: effect of cyclooxygenase and lipoxygenase inhibition evaluated by electron microscopical morphometry. AB - Leukocytes can take part in an inflammatory response in the heart after myocardial infarction or cardio-thoracic surgery. To investigate the injurious mechanism of activated polymorphonuclear leukocytes (PMN), isolated rat hearts were perfused with phorbol 12-myristate 13-acetate (PMA) activated PMN (3 x 10(6)/ml) alone for 10 min, in combination with a mixture of oxygen free radical scavengers (superoxide dismutase+catalase+thiourea) or in combination with ibuprofen (IBU), a cyclooxygenase inhibitor or diethylcarbamazine (DCM), a lipoxygenase inhibitor or BW 755C, a dual inhibitor of cyclooxygenase and lipoxygenase and an oxygen free radical scavenger. After 30 min of recovery, the hearts were perfusion-fixed with glutaraldehyde for electron microscopical examination. Based on examination of 25 micrographs per heart obtained by a random sampling procedure and on morphometric methods, volume fractions (Vv) of mitochondria (mito), altered mitochondria (alt mito), myofilament, and cellular edema were measured as fractions of myocyte volume. The most important finding was that Vv(alt mito/myocyte) was 0.09 +/- 0.16 and 0.02 +/- 0.04 in the hearts receiving PMN+PMA alone and when scavengers were added, respectively, whilst no changes in mitochondrial ultrastructure was observed after addition of IBU, BW 755C or DCM. Vv(mito/myocyte) was for PMN+PMA alone: 0.33 +/- 0.04, +scavengers: 0.29 +/- 0.02 +IBU:0.29 +/- 0.02, +BW 755C: 0.23 +/- 0.03*, +DCM: 0.28 +/- 0.02 (mean +/- S.D., *P < 0.05 compared to PMN+PMA). Capillary wall volume (cap wall) as a fraction of the whole capillary was also quantified. Vv(cap wall/cap) was for PMN+PMA alone: 0.26 +/- 0.06, +scavengers: 0.22 +/- 0.03, +IBU: 0.19 +/- 0.04*, +BW755C: 0.21 +/- 0.03, +DCM: 0.15 +/- 0.04* (*P < 0.05). These results further strengthen the notion that activated PMN are intravascularly active. In addition to exerting a cardiodepressive effect the present study shows that activated PMN can induce structural changes in the heart through the combined action of oxygen free radicals and arachidonic acid metabolites. PMID- 8729064 TI - Alterations of performance and oxygen utilization in chronically infarcted rat hearts. AB - Progressive dilatation of left ventricle has been demonstrated in hearts post infarction. However, the relationship of performance and energy consumption in chronically infarcted heart has not been clarified. To address this problem, we measured left ventricular pressure and oxygen consumption (MVO2) during stepwise increases in left ventricular filling volume in isolated isovolumic buffer perfused rat hearts 8 weeks after let coronary artery ligation or sham-operation. Systolic pressure-volume area (PVA) was calculated as an estimate of total mechanical energy consumed by the heart. The MVO2-PVA relation was analysed to define the economy of the contractile machinery in surviving myocardium. Structural dilatation and reduced pressure generation in infarcted hearts were indicated by a rightward shift of pressure-volume curves and a reduced maximal developed pressure of the left ventricle (80 +/- 5 v 119 +/- 4 mmHg, P < 0.01) which was obtained at substantially higher left ventricular volume compared to control hearts (0.79 +/- 0.02 v 0.39 +/- 0.01 ml, P < 0.01). The slope of the MVO2-PVA relation was significantly lower in the infarcted compared to the control groups (1.02 +/- 0.16 v 1.44 +/- 0.10 10(-5) mlO2/mmHg/ml, P < 0.05), reflecting an increased efficiency of chemomechanical energy transduction in surviving myocardium. However, at the similar MVO2 ventricular pressure development was significantly lower in infarcted hearts due to the unfavorable geometry resulting from ventricular dilatation. PMID- 8729065 TI - L-propionylcarnitine enhancement of substrate oxidation and mitochondrial respiration in the diabetic rat heart. AB - This study was designed to determine whether L-propionylcarnitine (LPC) treatment is beneficial in preventing the depression in cardiac function from occurring in chronic diabetes. Diabetes was induced by tail vein injection of streptozotocin (60 mg/kg). Two weeks later, treatment was initiated by supplementing the drinking water with LPC at the concentration of 1 mg/ml. Following a 6-week treatment period, myocardial substrate utilization and cardiac function were determined in isolated working hearts. In a separate group of hearts, the effects of LPC treatment on mitochondrial respiration were also determined. The results showed that diabetic hearts, compared with those of controls, oxidized glucose at a much lower rate, but oxidized palmitate at a similar rate. The effect of diabetes compared a controls was also characterized by a pronounced decrease in cardiac pump function. Following treatment with LPC, however, there was a marked increase in the rates at which glucose and palmitate were oxidized by diabetic hearts, and a significant improvement in cardiac pump performance. In addition, the depression of cardiac mitochondrial respiration seen in diabetes was prevented with LPC treatment. Our findings show that the depression of cardiac pump function by diabetes can be prevented with chronic LPC treatment. Possible mechanisms for this beneficial effect include an energetically favorable shift in glucose and fatty acid metabolism. PMID- 8729066 TI - Ranolazine increases active pyruvate dehydrogenase in perfused normoxic rat hearts: evidence for an indirect mechanism. AB - Ranolazine has shown anti-anginal efficacy in humans and cardiac anti-ischaemic activity in models, but without affecting haemodynamics or baseline contraction. In isolated normoxic rat hearts, Langendorff-perfused for 30 min with 11 mM glucose, 3% albumin, and 0.4 mM or 0.8 mM palmitate, 20 microM ranolazine significantly increased active, dephosphorylated, pyruvate dehydrogenase (PDHa), but not with no palmitate or 1.2 mM palmitate. Dichloroactetate (DCA, 1 mM), a PDHa kinase inhibitor, significantly increased PDHa in hearts perfused with 0, 0.4 or 0.8 mM but not 1.2 mM palmitate. PDHa was significantly increased with 1.2 mM palmitate by DCA plus ranolazine, and additive effects were also seen at 0.8 mM palmitate. Activation of PDH by ranolazine and promotion of glucose oxidation offers a plausible means by which the drug may be anti-ischaemic nonhaemodynamically. Extensive studies with extracted enzymes and isolated rat heart mitochondria failed to demonstrate any effects of ranolazine on PDH kinase or phosphatase, or on PDH catalytic activity, whereas effects of other known effectors (such as DCA) were readily demonstrable, suggesting that ranolazine activates PDH indirectly. Further analyses of the hearts revealed that ranolazine reduced acetyl CoA content under all conditions where fatty acid was present, and +/- DCA which itself had little effect. In the absence of fatty acid, ranolazine and/or DCA raised acetyl CoA. In perfusions where octanoate (+/- albumin) replaced palmitate, ranolazine still decreased acetyl CoA, but not when acetate replaced palmitate. In octanoate-perfused hearts, the contents of the C4, C6 and C8 CoA esters were all increased by ranolazine. This is consistent with ranolazine causing an inhibition of fatty acid beta-oxidation leading to decreased acetyl CoA and activation of PDH. PMID- 8729067 TI - Characterization and hemodynamic implications of renal vascular angiotensin II receptors in SHR. AB - The intrarenal renin-angiotensin system (RAS) contributes to the increased renal vascular resistance and reactivity observed in spontaneously hypertensive rats (SHR) and to the pathogenesis of high blood pressure (BP). Thus, we decided to characterize angiotensin II (ANG II) receptors in the renal arteries and glomeruli of 16-week-old SHR and their age-matched, normotensive Wistar-Kyoto (WKY) controls. SHR had significantly higher BP (153 +/- 4 v 96 +/- 10 mmHg) and heart weight (440 +/- 5 v 327 +/- 4 g/100 g body weight) than WKY rats. There was no difference in plasma renin activity between strains. Radioligand binding assays using non-peptide antagonists for AT1 (losartan) and AT2 (PD 123319) showed that renal preglomerular microvessels and glomeruli expressed a single receptor population (AT1) for ANG II. AT1 density tended to be lower in glomeruli of SHR compared to WKY (377 +/- 45 v 555 +/- 74 fmol/mg protein), but was significantly higher in preglomerular vessels (93 +/- 7 v 57 +/- 1 fmol/mg protein). No difference in receptor affinity was found in either preparation. Isolated kidney perfusion revealed that at low flow (3-10 ml/min), perfusion pressure was similar in both strains; however, at higher flow levels, SHR showed higher reactivity and less compliance than their controls. In addition, SHR presented a higher renal vascular reactivity to ANG II (but not to arterenol) than WKY rats. Thus, upregulation of ANG II receptors in the renal vasculature may mediate the hyperreactivity to ANG II observed in SHR kidney. PMID- 8729068 TI - Lack of evidence for a role of mast cell degranulation in acute hypoxia/reoxygenation-induced injury in the isolated rat heart. AB - In the present study, we evaluated the potential role of mast cell degranulation in acute hypoxia/reoxygenation-induced injury to cardiomyocytes in the isolated rat heart. Histamine release was determined to delineate the extent of mast cell degranulation, whereas the release of creatine kinase (CK) and lactate dehydrogenase (LDH) was assessed to quantitate the extent of irreversible injury to cardiomyocytes. The suitability of peroxidase (PO) as a marker for mast cell degranulation was also evaluated. Reoxygenation resulted in a release of histamine corresponding with 6.5% +/- 0.6% of total tissue content, whereas LDH, CK and PO release amounted to 30% +/- 2%, 28% +/- 2% and 32% +/- 3% of their respective tissue contents. Identical perfusion in the presence of the mast cell stabilizer lodoxamide tromethamine resulted in a reduced histamine release (2.8% +/- 0.1%) of total tissue content upon reoxygenation, but the release of LDH, CK or PO was not influenced. Cumulative histamine release did not correlate with the amount of LDH, CK or PO released. Treatment with consecutive bolus injections of the mast cell degranulating compound 48/80 during normoxic perfusion resulted in an almost complete histamine release, whereas PO release remained below detection limit. When the compound 48/80-treated hearts were subjected to hypoxia/reoxygenation, the release of LDH, CK or PO during reoxygenation again remained unchanged, whereas histamine release was negligible. Determination of PO activity of freshly isolated cardiomyocytes demonstrated that the bulk of PO in rat hearts was located in this particular cell type. Therefore we conclude that in the isolated rat heart, PO release is not a specific marker of mast cell degranulation. In addition, our data provide no firm evidence that in this experimental model, mast cell degranulation contributes to a significant extent to acute hypoxia/reoxygenation-induced injury to cardiomyocytes. PMID- 8729069 TI - Oxidative stress as a mechanism of cardiac failure in chronic volume overload in canine model. AB - We investigated the effects of chronic volume overload in the absence or presence of vitamin E supplements on the cardiac function and contractility, cardiac malondialdehyde (MDA)--a lipid peroxidation product--cardiac antioxidant enzyme activity and antioxidant reserve in canine model. The dogs were divided into three groups of seven dogs each: group I, control; group II, mitral regurgitation (MR) of 4 months duration; and group III, MR of 4 months duration receiving vitamin E (40 U/kg/daily) orally. MR was created by detaching two or more chordae tendinae to raise left atrial pressure to 2.5 to three times normal. MR produced a decrease in the index of myocardial contractility with little change in myocardial function. Decrease in myocardial (left and right ventricles) contractility was associated with an increase in cardiac MDA, and a decrease in cardiac antioxidant reserve and antioxidant enzyme activity. Prevention of volume overload-induced decrease in myocardial contractility by vitamin E was associated with a decrease in cardiac MDA and an increase in cardiac antioxidant reserve and glutathione peroxidase activity towards control levels. Superoxide dismutase and catalase activity remained depressed in vitamin E-treated group. The results indicate that chronic volume overload decreases the contractility of both right and left ventricles and is associated with oxidative stress in both ventricles. These results support the hypothesis that oxygen free radicals are involved in the chronic volume overload-induced cardiac depression. PMID- 8729070 TI - Depressed transient outward current density in ventricular myocytes from cardiomyopathic Syrian hamsters of different ages. AB - We determined whether the dilated cardiomyopathy which develops between 30 and 140 days of age in the Syrian hamster strain MS200, before the onset of cardiac hypertrophy and failure, is associated with alterations in both the action potential (AP) and the Ca(2+)-independent transient outward current, Ito1. AP was recorded in perfused hearts using microelectrodes and Ito1 was recorded in single ventricular myocytes using the whole-cell patch-clamp. The MS200 strain was compared to the control CHF148 strain at different periods of age (60, 90, 120 and 180 days). APs were markedly lengthened in MS200 compared to CHF148 hearts at all ages studied. Cell membrane capacitance increased with age in the two strains, but was not significantly higher in MS200 than in CHF148 of a given age, indicating the absence of cell hypertrophy. At 60 days of age, Ito1 density was the same in the two strains. Later on, Ito1 density increased markedly at 90-120 days then decreased at 180 days in CHF148, whereas this increase was delayed and of reduced amplitude in MS200. The sustained component of outward current, Isus, was not sizeably different in the two strains. The conductance-voltage and steady state inactivation relationships were shifted with age towards positive potentials by 15 mV in the two strains, but earlier in MS200 (90 days) than in CHF148 (180 days). Similarly, the recovery of Ito1 from inactivation exhibited a slow component which increased with age in the two strains but was larger in MS200 than in CHF148. In conclusion, alterations of Ito1 may contribute to changes in shape of AP, but cannot entirely explain dilation-induced AP lengthening. PMID- 8729071 TI - Adenosine and carbachol are not equivalent in their effects on L-type calcium current in rabbit ventricular cells. AB - Adenosine is one of the most important inhibitory modulators of heart function, producing negative inotropic, chronotropic and dromotropic effects and is also a major regulator of coronary circulation. The decrease in contractility by adenosine is mediated through inhibition of adenylyl cyclase by Gi-proteins coupled to adenosine receptors. However, little is known about the developmental differences in the effect of adenosine on cardiac cells. We have now shown that there is a striking developmental difference in the inhibitory effect of adenosine on isoproterenol-stimulated ICa between adult and newborn rabbit ventricular cells. Adenosine had no significant inhibitory effect on 0.1 muM isoproterenol-stimulated ICa in adult cells, while it completely blocked the 10 muM isoproterenol-stimulated ICa in newborn cells with an inhibitory potency similar to carbachol in newborn cells. Similarly, adenosine did not decrease the isoproterenol-stimulated cAMP levels in adult cells while it inhibited isoproterenol-stimulated cAMP levels significantly and equipotently to carbachol in newborn. However, for forskolin-stimulated ICa and cAMP levels in newborn cells, adenosine had a much lower inhibitory potency than carbachol. In adult cells, forskolin-stimulated ICa and cAMP levels were not affected by adenosine. We showed previously that the Gia3 isoform of inhibitory G protein was present in newborn cell membranes, but not detectable in adult cell membranes. We have now used a synthetic decapeptide corresponding to the C-terminal sequence of Gia3 in the patch pipette and have shown a selective partial block of the inhibitory action of adenosine for isoproterenol-stimulated ICa, suggesting that the inhibitory action of adenosine on ICa is mediated primarily through the Gia3 pathway. PMID- 8729073 TI - Regional changes in angiotensin II receptor density after experimental myocardial infarction. AB - The plasma and cardiac renin-angiotensin systems may be activated after myocardial infarction. The myocardium may therefore be exposed to increased concentrations of angiotension II, which may contribute to myocardial injury. The purpose of this study was to identify the potential sites of action of angiotensin II in the infarcted heart. Myocardial infarction was induced in rats by left coronary artery ligation, and the hearts were removed for study after 18 h, 7 days, or 8 months. The regional ventricular angiotensin II receptor density was assessed by [125I](Sar1,Ile8)angiotensin II binding and quantitative autoradiography. The [125I](Sar1,Ile8)angiotensin II binding was unchanged at 18 h, but was increased at 7 days in the infarcted region of the left ventricle (73.2 +/- 3.2 amol/mm2, mean +/- S.E.M.) compared with the non-infarcted region (1.6 +/- 0.2 amol/mm2, P < 0.0001) and with the left ventricular myocardium of sham-operated control animals (1.3 +/- 0.1 amol/mm2, P < 0.0001). The increased [125I](Sar1,Ile8)angiotensin II binding density was still present, but diminished, at 8 months after coronary ligation (49.0 +/- 5.7 amol/mm2, P < 0.0001 v control, P = 0.0058 v 7-day infarcts). The increased binding of [125I](Sar1,Ile8)angiotensin II was antagonised by losartan, an AT1 receptor antagonist, but not by an AT2 receptor antagonist. Microautoradiography of [125I](Sar1,Ile8) angiotensin II, and assessment of collagen deposition using picrosirius staining and immunostaining demonstrated that the regional increase in AT1 receptor density in the infarcted region of myocardium was associated with fibroblast infiltration and collagen deposition. The infarct scar and the cardiac fibroblasts within it express high levels of angiotension II receptors and therefore represent potential targets for the actions of angiotensin II after myocardial infarction. PMID- 8729072 TI - Effect of ischemic preconditioning on mitochondrial oxidative phosphorylation and high energy phosphates in rat hearts. AB - The ability of ischemic preconditioning (IP) to protect the myocardium against prolonged ischemia may derive from improved energy balance. We therefore examined myocardial energy metabolism and mitochondrial oxidative phosphorylation in isolated perfused rat hearts which were either subjected (IP group), or not subjected (control group), to preconditioning prior to 30 min sustained ischemia and 30 min reperfusion. Preconditioning was achieved with two cycles of 5 min ischemia followed by 5 min reperfusion. Recovery of myocardial function was significantly greater, and creatine kinase release was significantly lower, in the IP group. Although ATP hydrolysis during the sustained ischemia remained unchanged in both groups, greater preservation of high energy phosphate (eg. ATP and CP) was observed in the IP group after reperfusion. CP content immediately after preconditioning greatly exceeded pre-ischemic values. Lactate production during the sustained ischemia was significantly lower in the IP group, suggesting a decrease in anaerobic glycolysis and a probable attenuation of intracellular acidosis. Oligomycin-sensitive mitochondrial ATPase activity in the control group was significantly decreased both after the sustained ischemia and the reperfusion, but in the IP group it did not change after the preconditioning, sustained ischemia, or reperfusion. Although atractyloside-inhibitable adenine nucleotide translocase activity was markedly decreased during sustained ischemia in both groups, its activity was significantly higher after reperfusion in the IP group. These data suggest that (1) mitochondrial ATPase contributes only slightly to ATP depletion during sustained ischemia, (2) both the CP overshoot phenomenon and the decrease in anaerobic glycolysis can be attributable to cardioprotection during the sustained ischemia, and (3) the preservation of ATPase and adenine nucleotide translocase activities may be a possible explanation for the restoration of high energy phosphates after sustained ischemia-reperfusion injury in the preconditioned hearts of rats. PMID- 8729074 TI - Balloon angioplasty significantly increases collagen content but does not alter collagen subtype I/III ratios in the atherosclerotic rabbit iliac model. AB - Angioplasty has been reported to increase collagen content and to alter vascular collagen alpha 1(I), alpha 2(I), and alpha 1(III) mRNA levels. Collagen synthesis is tightly regulated by complex translational and post-translational mechanisms such that mRNA levels may not necessarily reflect biosynthesis. To test whether collagen subtype I and III protein levels are altered by angioplasty, we quantitatively analysed collagen I/III protein ratios at 4 weeks after balloon angioplasty. Twenty New Zealand White rabbits underwent iliac artery balloon de endothelialization and then were placed on a 0.5% cholesterol/6% peanut oil diet for 7 weeks at which time angioplasty was performed on arteries with > or = 50% stenosis. Arteries with < 50% stenosis were not dilated and served as controls. Animals were killed 4 weeks later and hydroxyproline (OH-pro) content and subtype I/III ratios were analysed in 5-mm mid-iliac sections. OH-pro was measured by a colorimetric assay. Subtype ratios were determined by a highly quantitative two dimensional cyanogen bromide peptide mapping method. The degree of stenosis was measured as the minimal vessel lumen diameter and calculated as a percentage stenosis compared to a proximal reference segment. Calculated collagen content (micrograms/mg tissue) was significantly higher 4 weeks following angioplasty compared to the non-dilated group (220.4 +/- 70.8 v 308.2 +/- 26.9, P = 0.04; n = 12), despite similar percentage stenosis in the primary and restenotic lesions. The ratio of collagen I/III subtype protein distribution was not significantly different in the non-dilated and angioplastied groups (4.88 +/- 1.00 v 4.70 +/- 0.82, respectively). These studies are the first to provide data on collagen I/III subtypes following angioplasty and suggest that collagen accumulation may be more important in restenosis than alteration of collagen protein subtypes. PMID- 8729075 TI - Cytomegalovirus in the principal submandibular gland of the little brown bat, Myotis lucifugus. AB - In the course of a large scale study of salivary gland ultrastructure in chiropterans, enlarged cells infected with numerous virus particles were encountered in some acinar cells in the principal submandibular glands of two of 34 little brown bats (Myotis lucifugus). The characteristic morphology of the viruses, together with the cytomegaly that they induced, led to their identification as cytomegalovirus (CMV). In a reversal of the situation in other animal species, bat virus particles within cytoplasmic vacuoles lacked capsomeres, whereas the latter were prominent in particles free in the cytosol. The generally accepted schemes for CMV production cannot explain this seemingly aberrant morphology. This report extends to four the number of mammalian orders in which CMV has been documented by means of electron microscopy as occurring in salivary glands. PMID- 8729076 TI - Comparison of the effects of infectious bronchitis and infectious laryngotracheitis on the chicken respiratory tract. AB - In infectious bronchitis (IB) virus infection of the chicken the upper and lower respiratory tracts were damaged, but infectious laryngotracheitis (ILT) virus caused lesions only in the upper respiratory tract. Secondary infection with Escherichia coli was apparent in the trachea of birds inoculated with either virus but was more striking in those given IB virus. Serum alpha 1-acid glycoprotein, an acute-phase protein, occurred in higher concentrations in chickens inoculated with IB virus than in those given ILT virus. PMID- 8729077 TI - Histopathological changes in the lymphoid tissues of sheep exposed to the bovine immunodeficiency-like virus. AB - Six yearling sheep were inoculated intraperitoneally with peripheral blood from two sheep infected with the bovine immunodeficiency-like virus (BIV) strain R29. An additional five sheep served as sham-inoculated controls. Of the six sheep given BIV, five seroconverted, one of them remaining seropositive for the duration of the study. The polymerase chain reaction demonstrated BIV provirus in three of the five serologically positive animals. At necropsy, 1 year after inoculation, histological changes were found only in the lymphoid tissues. In sheep exposed to BIV, mild though significant increases were seen in the (1) number of splenic periarteriolar lymphoid sheaths, (2) number of secondary follicles in hilar and prescapular or popliteal lymph nodes, and (3) medullary sinus cellularity in prescapular and popliteal lymph nodes. PMID- 8729078 TI - Expression of interleukin-1 beta, interleukin-6 and tumour necrosis factor alpha by macrophages in canine lymph nodes with mineral-associated lymphadenopathy, granulomatous lymphadenitis or reactive hyperplasia. AB - The distribution of CD3+ T lymphocytes and the expression of interleukin-1 beta (IL-1 beta), interleukin-6 (IL-6) and tumour necrosis factor alpha (TNF alpha) by macrophages in canine lymph nodes with mineral-associated lymph-adenopathy, granulomatous lymphadenitis or reactive hyperplasia was examined immunohistochemically. In areas of preserved lymph node architecture, CD3+ T lymphocytes were located in the paracortex, with scattered cells in the follicular mantle and germinal centre, and small aggregates in the medullary cords. T cells were prominent within areas of diffuse granulomatous inflammation and formed a peripheral rim around microgranulomas. TNF alpha expression was demonstrated in macrophages from seven of 11 dogs with mineral-associated lymphadenopathy, one of five dogs with granulomatous lymphadenitis and three of five with reactive hyperplasia. Positive macrophage staining for IL-1 beta and IL 6 was seen in two lymph nodes with reactive hyperplasia. Lymph nodes from control (clinically normal) dogs showed no evidence of cytokine expression. Neutrophils expressing IL-1 beta were observed within the medullary sinuses of two lymph nodes with mineral-associated lymphadenopathy and one with reactive hyperplasia, and in the inflammatory infiltrate in one lymph node with granulomatous lymph adenitis. The sensitivity of immunohistochemical examination as a means of detecting cytokines in abnormal tissue is discussed. PMID- 8729079 TI - Neural cells from dogs with spontaneous distemper encephalitis express class II major histocompatibility complex molecules. AB - Expression of class II major histocompatibility complex (MHC) molecules by non immune cells (e.g., parenchymal cells) leads to the presentation of self antigens, and may have a role in the pathogenesis of many diseases mediated by autoimmunity. Such diseases, characterized by demyelination of the central nervous system and expression of class II MHC molecules on neural cells, include multiple sclerosis, experimental allergic encephalitis and Theiler's murine encephalomyelitis virus infection. Canine distemper encephalitis probably does not have an autoimmune character, but it shares many similarities with the aforementioned diseases. For this reason, the expression of class II MHC molecules in the brains of dogs with canine distemper encephalitis was investigated immunohistochemically. The results presented here demonstrate that canine microglia and astrocytes "upregulate" class II MHC expression in cases of encephalitis associated with chronic canine distemper. PMID- 8729080 TI - Trypanosoma congolense infection in sheep: cellular phenotypes in lymph and lymph nodes associated with skin reactions. AB - Intradermal inoculation of sheep with culture-derived metacyclic forms of Trypanosoma congolense resulted in the development of localized skin reactions (chancres) and enlargement of the draining lymph nodes 7 days after infection. Changes in the expression of surface antigens of lymphocytes in lymph leaving the affected skin reactions and in the associated lymph nodes were monitored by cannulating the afferent and efferent lymphatic ducts. Trypanosomes appeared in afferent and efferent lymph 3 to 5 days after infection and persisted even as the chancres regressed. The cellular output in both afferent and efferent lymph increased markedly after the onset of parasitosis. Sequential analysis of the phenotypes of lymphocytes by immunofluorescent staining and flow cytometry revealed that in afferent lymph draining the chancre there was an early response which was due to an increase in T cells, particularly CD4+ and CD8+ cells; however, as the chancres-regressed there was an increase in lymphoblasts and surface immunoglobulin-bearing cells. In contrast, in the efferent lymph, the increase in lymphocytes was due predominantly to a higher number of cells bearing surface immunoglobulins. PMID- 8729081 TI - Newcastle disease virus-induced apoptosis in the peripheral blood mononuclear cells of chickens. AB - Specific pathogen-free chickens were inoculated with the GB strain of Newcastle disease virus by the ocular route, and were killed at daily intervals after infection. Peripheral blood mononuclear cells (PBMCs) were harvested for DNA extraction, electron microscopy, and flow cytometry studies to detect the presence of cellular apoptosis and necrosis. Extracted DNA showed fragmentation at 4 and 6 days after infection. Electron microscopy showed that the chromatin of the lymphocytes became condensed and occasionally crescent-shaped; apoptotic bodies could also be seen. Some heterophils became darkly stained and their nuclei degenerated; others seemed to have lost their cytoplasmic membrane, but had intact granules and condensed chromatin. Flow cytometry studies showed that the number of normal PBMCs gradually decreased from 99.04% at the beginning of the infection to 45.24% on the sixth day of infection, with a concomitant increase of apoptotic cells. The results show that PBMCs of chickens exhibit typical signs of apoptosis after infection by the Newcastle disease virus. PMID- 8729082 TI - Ultrastructural changes in the cardiac muscle of chickens infected with the GB strain of Newcastle disease virus. AB - The changes in chicken cardiomyocytes resulting from infection with the GB strain of Newcastle disease virus were studied by electron microscopy. The virus infection caused shortening of the sarcomeres and generalized oedema in the perivascular region of the heart muscle, containing debris from degenerated mitochondria and myofilaments. Some of the cardiomyocytes had condensed chromatin, resembling that of apoptotic cells. PMID- 8729083 TI - Relationship of acid phosphatase activity to ultrastructural features in mice inoculated with Mycobacterium paratuberculosis. AB - Macrophage activation, measured as increased acid phosphatase (AcPase)-positive areas by image analysis, and ultrastructural features were examined in granulomatous mycobacterial lesions of mice innately susceptible (BALB/c mice; Bcg) and innately resistant (C3H/HeJ mice; Bcg) to Mycobacterium paratuberculosis strain ATCC 19698. In the liver and spleen of BALB/c mice 3 weeks after intraperitoneal inoculation with M. paratuberculosis, AcPase activity detected in epithelioid cell nodules was high; it had decreased, however, in the liver and spleen after a further 3 and 6 weeks, respectively. In C3H/HeJ mice, the size of epithelioid cell nodules in the liver and spleen was smaller than in BALB/c mice, and infiltrating macrophages, which had increased by week 9 after inoculation, showed high AcPase activity. Ultrastructurally, by week 32 in BALB/c mice, small phagolysosomes (SPLs) had greatly increased in number in the epithelioid cells. These SPLs contained a few AcPase-positive areas and a small number of bacteria, most of which were surrounded by an electron-translucent space (or electron transparent zone [ETZ]). In contrast, only a few SPLs were observed in C3H/HeJ mice at week 32; in the liver and spleen, large phagolysosomes (LPLs) showed high AcPase activity and contained many degenerated bacteria, which also had an ETZ. These results suggest that the enzymatic and ultrastructural differences in phagolysosomes between BALB/c mice and C3H/HeJ mice reflect the susceptibility of these mouse strains to M. paratuberculosis. PMID- 8729084 TI - Dual infection with attaching and effacing Escherichia coli and enterotoxigenic Escherichia coli in post-weaning pigs. AB - Post-weaning diarrhoea in pigs occurred on two farms in Hokkaido, Japan, in 1994. Four piglets aged 35 or 45 days were examined after death. At necropsy, ecchymotic haemorrhages were seen on the mucosal surface of the caecum and colon. Histopathologically, numerous Gram-negative bacilli adhered to the brush border of the small intestines, but the brush border itself was intact. Typical attaching and effacing (AE) lesions were seen in the caecum and colon. Immunohistochemically, the bacilli which adhered to the brush border gave positive results with antisera against serogroup O149 of Escherichia coli; the bacilli which caused the AE lesions, however, belonged to serogroup O45. It was concluded that the disease resulted from dual infection with attaching and effacing E. coli (AEEC) and enterotoxigenic E. coli (ETEC). PMID- 8729085 TI - Skin tumours in cattle and sheep after freeze- or heat-branding. AB - This article describes the occurrence of carcinogenic epidermal neoplasia (papillomas and squamous cell carcinoma) at the sites of tattooing with liquid nitrogen in dairy cattle, and squamous cell carcinoma at the sites of freeze- or heat-branding in beef cattle and sheep. PMID- 8729086 TI - A logic of disease. PMID- 8729087 TI - Polyunsaturated fatty acid regulation of hepatic gene transcription. AB - Polyunsaturated fatty acids (PUFA) of the n-6 and n-3 families inhibit transcription of a number of hepatic lipogenic and glycolytic genes, e.g. fatty acid synthase. In contrast, saturated and monounsaturated fatty acids exert no suppressive action on lipogenic gene expression. The unique PUFA regulation of gene expression extends beyond the liver to include genes such as adipocyte glucose transporter-4, lymphocyte stearoyl-CoA desaturase 2, and interleukins. Some of the transcriptional effects of PUFA appear to be mediated by eicosanoids, but PUFA suppression of lipogenic and glycolytic genes is independent of eicosanoid synthesis and appears to involve a nuclear mechanism directly modified by PUFA. With the recent cloning of a fatty acid-activated nuclear factor termed peroxisome-proliferator-activated receptor (PPAR) has come the suggestion that PPAR may be the PUFA response factor. This review, however, presents several lines of evidence that indicate that the PPAR and n-6 and n-3 PUFA regulation of lipogenic and glycolytic gene transcription involve separate and independent mechanisms. Thus PPAR appears not to be the PUFA response factor. PMID- 8729088 TI - Effect of polyunsaturated fatty acids in obese mice. AB - Genetically obese (ob/ob) mice display a variety of metabolic differences from lean litter mates. In the obese state, fatty acid desaturation-elongation in brown adipose tissue mitochondria is apparently altered, resulting in differences in membrane fatty acid composition. This change in membrane lipid environment appears to influence GDP binding and therefore the activity of the proton conductance pathway associated with regulation of energy expenditure in these animals. In liver, binding of insulin to the nuclear membrane is increased by feeding a high polyunsaturated/saturated (P/S) diet fat. Consumption of a high P/S diet decreased mRNA levels for fatty acid synthase, acetyl-CoA carboxylase, malic enzyme, and pyruvate kinase in obese and lean animals. Expression of mRNA for these lipogenic enzymes was higher in obese animals and suggests that obese mice may be resistant to polyunsaturated fatty acid feedback control of gene expression. PMID- 8729089 TI - Dietary marine lipids suppress continuous expression of interleukin-1 beta gene transcription. AB - n-3 Polyunsaturated fatty acids abundant in marine lipids suppress certain inflammatory and immune reactions, and dietary marine lipid supplements have antiinflammatory effects in experimental and human autoimmune disease. Previous work by other investigators demonstrated that dietary marine lipid supplements suppressed production of cytokines from stimulated human peripheral blood mononuclear cells ex vivo. The present study further documents the ability of n-3 fatty acids to inhibit cytokine formation, and in part defines the mechanism of the inhibition of production of interleukin-1 beta (IL-1 beta) by dietary n-3 fatty acid. Female BALB/c mice were each fed a fat-free balanced diet to which was added either a refined fish oil (FO) preparation as a source of n-3 fatty acid, or beef tallow (BT), which consisted primarily of saturated and monoenoic fatty acids. After ingesting the experimental diets for periods ranging from 3 to 12 wk. spleen cell preparations were stimulated ex vivo with either lipopolysaccharide (LPS) or phorbol 12-myristate 13-acetate (PMA), and proIL-1 beta mRNA (IL-1 beta mRNA) was measured by northern analysis. Levels of IL-1 beta mRNA in both LPS- and PMA-stimulated cells from BT-fed mice were elevated to a greater extent than in cells from FO-fed mice, at most concentrations of LPS and PMA. Stability of LPS-stimulated mRNA levels after actinomycin D was similar for BT and FO groups, indicating that lower levels of IL-1 mRNA with FO groups was related to suppressed IL-1 gene transcription and not due to accelerated transcript degradation. Nuclear run-on transcription assays revealed a more transient expression of the IL-1 beta gene in LPS-stimulated spleen cells from FO fed mice compared to cells from BT-fed mice. We conclude that dietary marine lipids reduce transient expression of the IL-1 beta gene in stimulated splenic monocytic cells. Preliminary results from nuclear run-on transcription assays indicate that n-3 fatty acids may not change the initial rate of gene transcription but may promote more rapid shutting down of transcription of this gene after induction than do alternative lipids. PMID- 8729090 TI - Polyunsaturated fatty acids inhibit hepatic stearoyl-CoA desaturase-1 gene in diabetic mice. AB - Insulin and dietary fructose independently induce stearoyl-CoA desaturase 1 (SCD1) gene expression in diabetic mouse liver. In the present study, we again used diabetic mice and supplemented a high fructose diet with polyunsaturated fatty acids (PUFA) to determine the selective repression of SCD1 gene expression by dietary PUFA, as previously shown in normal mice. We saw dramatic repression of SCD1 mRNA expression, with trilinolenin at 3% and triarachidonin at 1% supplementation. We also observed significant repression of insulin-induced SCD1 mRNA upon supplementation of the noninducing starch diet with PUFA. In conclusion, our data demonstrate that PUFA negatively regulate hepatic gene expression through an insulin-independent mechanism. PMID- 8729091 TI - Effects of different polyunsaturated fatty acids on growth-related early gene expression and cell growth. AB - Effects of polyunsaturated fatty acids on expression of early-response genes c fos and Egr-1 and induction of cell growth were assessed in Swiss 3T3 fibroblasts. Stimulation with arachidonic acid increased mRNA levels of c-fos and Egr-1. This effect was inhibited by preincubation with cyclooxygenase inhibitors and restored by addition of prostaglandin E2 (PGE2), the predominant eicosanoid produced in Swiss 3T3 fibroblasts. Further signaling of PGE2, was mediated by a protein kinase C-dependent pathway, since downregulation, or inhibition, of protein kinase C reduced increases in mRNA levels. Parallel to the stimulatory effects on mRNA levels, AA and PGE2 also increased cell growth, as determined by uptake of [3H]-thymidine. In contrast to arachidonic acid, n-3 fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) did not increase c-fos and Egr-1 mRNA levels or cell growth. Furthermore, preliminary data indicate that EPA and DHA even reduce the stimulatory effect of AA, which is associated with reduced formation of PGE2. In conclusion, our data indicate that AA increases expression of growth-related early genes c-fos and Egr-1 Swiss 3T3 fibroblasts by its conversion to PGE2 and subsequent activation of protein kinase C, whereas n-3 fatty acids do not activate this signaling cascade. PMID- 8729092 TI - The cardiac antiarrhythmic effects of polyunsaturated fatty acid. AB - Each year in the United States alone some 250,000 persons die within one hour of an acute myocardial infarction. These deaths are largely due to ischemia-induced ventricular arrhythmias, primarily ventricular fibrillation (VF). Thus a safe, simple means of preventing such arrhythmias has considerable public health benefit potential. We have demonstrated that the intravenous infusion of n-3 polyunsaturated fatty acids (PUFA) from fish oils will prevent ischemia-induced VF in prepared, nonanesthetized, exercising dogs, confirming earlier feeding studies in rats. We show that this protective effect is due to an action of the free acidic form of the PUFA to alter the electrophysiology of individual cardiac myocyte so that the cells are electrically more stable. The electrophysiologic effects, in turn, result from direct and specific effects of the PUFA to block the fast voltage-dependent sodium channels. The binding of the free fatty acids is directly to the protein of the sodium channels and results in prolongation of the inactivated state of these channels. Other ion channels are also affected by the PUFA. Two clinical trials with n-3 PUFA are mentioned which inadvertently support the antiarrhythmic potential of PUFA ingestion. PMID- 8729093 TI - Eicosapentaenoic acid is primarily responsible for hypotriglyceridemic effect of fish oil in humans. AB - The aim of this study was to determine whether eicosapentaenoic acid (EPA) or docosahexaenoic acid (DHA), or both, were responsible for the triglyceride (TG) lowering effects of fish oil. EPA (91% pure) and DHA (83% pure), a fish oil concentrate (FOC; 41% EPA and 23% DHA) and an olive oil (OO) placebo (all ethyl esters) were tested. A total of 49 normolipidemic subjects participated. Each subject was given placebo for 2-3 wk and one of the n-3 supplements for 3 wk in randomized, blinded trials. The target n-3 fatty acid (FA) intake was 3 g/day in all studies. Blood samples were drawn twice at the end of each supplementation phase and analyzed for lipids, lipoproteins, and phospholipid FA composition. In all groups, the phospholipid FA composition changed to reflect the n-3 FA given. On DHA supplementation, EPA levels increased to a small but significant extent, suggesting that some retroconversion may have occurred. EPA supplementation did not raise DHA levels, however. FOC and EPA produced significant decreases in both TG and very low density lipoprotein (VLDL) cholesterol (C) levels (P < 0.01) and increases in low density lipoprotein (LDL) cholesterol levels (P < 0.05). DHA supplementation did not affect cholesterol, triglyceride, VLDL, LDL, or high density lipoprotein (HDL) levels, but it did cause a significant increase in the HDL2/HDL3 cholesterol ratio. We conclude that EPA appears to be primarily responsible for TG-lowering (and LDL-C raising) effects of fish oil. PMID- 8729094 TI - A new relationship between total/high density lipoprotein cholesterol and polyunsaturated fatty acids. AB - Dietary and plasma fatty acids have been linked to total cholesterol but not to the ratio of total/high-density lipoprotein cholesterol (TC/HDLC). To evaluate the relationship between dietary and plasma levels of polyunsaturated fatty acids (PUFA) and TC/HDLC, we analyzed cross-sectional and longitudinal data using 519 plasma samples (50% men, 50% women) from subjects participating in the Framingham Heart Study and results from a study feeding diets rich in either n-6 linoleic acid or n-3 alpha-linolenic acid with or without fish oil supplements (n-3 derivatives). Values of TC/HDLC are inversely related to the percent of plasma PUFA when both variables are measured at the same time in different subjects, R = 0.82, P < 0.000001. PUFA in phospholipids increase in response to increased dietary intake of different PUFA, either n-3 or n-6 or fish oils. There was a highly significant inverse relationship between TC/HDLC and the percent of PUFA in phospholipids, R = 0.97, P < 0.001. The relationship was similar regardless of the source and type of dietary fatty acids. A similar relationship existed when only the baseline points were considered. When plasma PUFA % increases, either in response to a diet high in PUFA or across different subjects, TC/HDLC ratios decline. Evaluation of plasma fatty acid profiles and increased balanced dietary intake of PUFA to bring fatty acid profiles of subjects with low PUFA plasma levels closer to the profile of a healthy reference group is an effective approach to reduce high TC/HDLC. Reductions of more than 50% in TC/HDLC appear feasible with dietary modification alone. Further research, into fatty acid metabolic activity may determine the biochemical basis of common dyslipidemias. PMID- 8729096 TI - Fats for the food industry: implications for cholesterol-lowering. AB - The special needs of the food industry may not always coincide with what is healthiest for consumers. People always demand optimal taste, flavor and texture; they expect shelf-stability, convenience and novelty. The edible oils and fat industry has been particularly vulnerable in this regard but has tried, often very successfully, to modify its products to take into account the customer's preferences and their health. The following review illustrates some potential strategies in the retail and commercial sectors, especially in baking, frying and food service areas. Given the large increase in the consumption of convenient, small portion meals, the fat content and composition of such meals and products will critically influence further outcomes in areas related to cardiovascular disease, cancer, obesity and diabetes. PMID- 8729095 TI - Control of endothelial leukocyte adhesion molecules by fatty acids. AB - Dietary balance of long-chain fatty acids (FA) may influence human susceptibility to pathological processes which involve the interaction of leukocytes with vascular endothelium, such as atherogenesis and inflammation. Such interaction is largely mediated by the de novo or increased expression of endothelial leukocyte adhesion molecules on vascular endothelial cells, able to tether and stably bind leukocytes onto the vessel wall, and by the production of leukocyte chemoattractants. Endothelial cells do not normally support high levels of leukocyte adhesion. They do so, however, when exposed to a number of stimuli, such as oxidized low density lipoprotein bacterial lipopolysaccharides, and inflammatory cytokines, which induce phenotypic changes generally referred to as "endothelial activation." We compared various FA in their ability to modulate endothelial activation by cytokines. FA included linoleic, arachidonic, oleic, eicosapentaenoic and, docosa-hexaenoic acid (DHA) as representatives of the n-6, n-3 polyunsaturated FA and of the monounsaturated FA. The n-3 FA DHA, and, to a lesser extent, oleate, at nutritionally compatible concentrations, were able to reduce endothelial expression of Vascular Cell and Adhesion Molecule-1 (VCAM-1). In further studies, DHA dose- and time-dependently reduced also the expression of E-selectin, Intercellular Adhesion Molecule-1, interleukin (IL)-6 and IL-8, in response to IL-1, IL-4, tumor-necrosis factor, or bacterial endotoxin. The magnitude of this effect paralleled its incorporation into cellular phospholipids. Also, coordinate with reduced surface adhesion molecule expression, DHA reduced the adhesion of human monocytes and of monocytic U937 cells to cytokine-stimulated endothelial cells. These effects were accompanied by a quantitatively consistent reduction in VCAM-1 mRNA, indicating a pretranslational control of adhesion molecule gene expression. These novel properties of FA as modulators of endothelial activation may help to explain the influence of dietary FA intake on atherogenesis and inflammation. PMID- 8729097 TI - Factors affecting resistance of low density lipoproteins to oxidation. AB - Oxidation resistance (OR) of low density lipoproteins (LDL) is frequently determined by the conjugated diene (CD) assay, in which isolated LDL is exposed to CU2+ as prooxidant in the range of 1-10 microM. A brief review on major findings obtained with this assay will be given. A consistent observation is that vitamin E supplements or oleic acid-rich diets increase OR. Oxidation indices measured by the CD assay and effects of antioxidants very significantly depended on the Cu2+ concentration used for LDL oxidation. For medium and high Cu2+ concentrations, the relationship between lag time and propagation rate can be described by a simple hyperbolic saturation function, which has the same mathematical form as the Michaelis-Menten equation. At medium and high Cu2+ concentrations (0.5 to 5 microM), vitamin E increases lag time in a dose dependent manner. The increase is higher for 0.5 microM Cu2+ as compared to 5 microM. At low Cu2+ concentrations (0.5 microM or less), the mechanism of LDL oxidation changes. Significant oxidation occurs in a preoxidation phase, which commences shortly after addition of Cu2+. Preoxidation is not inhibited by vitamin E. It is concluded that much additional work is needed to validate the importance of oxidation indices derived from CD and similar assays. PMID- 8729098 TI - Determinants of antioxidant status in humans. AB - Antioxidant status in humans reflects the dynamic balance between the antioxidant system and prooxidants and has been suggested as a useful tool in estimating the risk of oxidative damage. This paper reviews determinants of antioxidant status such as diet including antioxidant nutrient and nonnutrient intake, absorption and bioavailability, dietary components such as polyunsaturated fatty acids and transition metals, food storage and processing, chemical form, chirality and formulation of supplemental compounds and alcohol intake; environmental factors such as pollutants, ultraviolet radiation and smoking; injury and disease, medications and other medical treatments such as radiation; strenuous exercise; and physiological stage or conditions such as those in premature babies and the elderly. It is proposed that, in addition to current focus on tissues, the antioxidant status of digesta should be considered because of its effect on specific tissues and potential health implications. PMID- 8729099 TI - Antioxidants, lipoprotein oxidation, and arterial function. AB - Lipoproteins have an important influence on arterial tissues. High levels of low density lipoproteins (LDL) may inhibit arterial function in terms of the release of nitric oxide from the endothelium. This inhibition is enhanced by the oxidation of these lipoproteins which may occur during the development of atherosclerosis. Many of these effects are due to lipid oxidation products. The impairment of the release of nitric oxide and vasodilatation may be partially attenuated by antioxidants. LDL may also enhance the activity of blood platelets, especially when they are mildly oxidized. Oral administration of antioxidants may decrease the activation of platelets to a limited extent. Lipid peroxides may play a role in these processes. PMID- 8729100 TI - The effect of modest vitamin E supplementation on lipid peroxidation products and other cardiovascular risk factors in diabetic patients. AB - Among many factors, elevated lipids and lipid peroxide levels in blood are major risk factors in the development of cardiovascular disease in diabetic patients. This study has examined whether oral supplementation of vitamin E, an antioxidant, has any effect on blood lipid peroxidation products (LP) and lipid profile of diabetic patients. Thirty-five diabetics(D) were supplemented with DL alpha-tocopherol (E) capsule (orally, 100 IU/d) or placebo (P) for three months in double-blind clinical trials. Plasma E was analyzed by HPLC and LP by the thiobarbituric acid-reactivity; serum lipids by auto-analyzer. Data were analyzed using paired t-test and Wilcoxon Signed Rank Test. Vitamin E supplementation significantly lowered LP and lipid levels in diabetic patients; there were no differences in these parameters after P supplementation. There were no differences in the duration of diabetes and ages of D between P- and E- supplemented groups. This study suggests that vitamin E supplementation significantly lowers blood LP and lipid levels in diabetic patients. PMID- 8729101 TI - Modulation of antioxidant enzymes and programmed cell death by n-3 fatty acids. AB - Studies from our laboratory indicate that n-3 (fish oil, FO) lipids at 10% (w/w) in a nutritionally adequate, semipurified diet, and supplemented with equal levels of antioxidants, extended the life span of lupus-prone (NZB/NZW)F1 (B/W) female mice as compared to n-6 (corn oil, CO) lipids. The early rise of autoimmune disease in CO-fed mice was closely linked to the loss of T-cell function. Both IL-2 production and IL-2 receptor expression were reduced due to the loss of naive T-cells and a rise in memory T-cells. Proliferative response to both mitogens and superantigens (staphylococcal enterotoxins A and B) was higher in FO-fed 6.5-mon-old mice. These changes paralleled decreased PGE2 production by splenic cells from FO-fed mice. Analysis of mRNA expression in different organs revealed differential effects of dietary lipids. In FO-fed mice, transforming growth factor beta 1 (TGF beta 1) expression was decreased in kidneys, but splenic tissues had higher expression of TGF beta mRNA. As TGF beta promotes programmed cell death (PCD), we studied the effects of CO and FO on PCD rates in lymphocytes. Both propidium iodide staining and DNA fragmentation were elevated in lymphocytes of FO-fed mice when compared to CO-fed mice of similar age. Also, increased PCD correlated closely with increased Fas gene expression. Thus, in addition to various other antiinflammatory effects, dietary FO appears to increase PCD and prevent accumulation of self-reactive immune cells in lymphoid organs. Further studies are required to dissect the pro- and antiinflammatory mechanisms associated with dietary n-3 and n-6 lipids in modulating autoimmune disorders or malignancy during aging. PMID- 8729102 TI - In vivo labeling of brain phospholipids by long-chain fatty acids: relation to turnover and function. AB - An experimental method and model are described to quantitate kinetics of in vivo incorporation of fatty acids (FA) into stable brain phospholipids. When a radiolabeled long-chain FA is injected intravenously in a rat, it rapidly equilibrates with brain FA-CoA, the precursor pool for phospholipids. As different labeled FA enter different sn positions of specific phospholipids, a combination of labels can be used to investigate roles of different phospholipids in brain function and structure. By taking into account dilution lambda of specific activity of brain FA-CoA, compared with specific activity of FA in plasma, half-lives of FA in individual brain phospholipids can be calculated. Values for lambda less than 0.02 suggest marked recycling, and give half-lives two orders of magnitude smaller than literature values. A half-life of arachidonate in phosphatidylinositol of 0.66 h (turnover = 105%h) is consistent with active participation of this FA in phospholipase A2 mediated signal transduction. PMID- 8729103 TI - Essential fatty acid uptake and metabolism in the developing rodent brain. AB - Studies were carried out to determine whether the brain takes up and metabolizes essential fatty acids during early postnatal development in rodents. Rats and mice were dosed with deuterium-labeled linoleic and linolenic acids either by intraperitoneal injection or by gavage. Animals were killed at different times thereafter, and organs were removed. Brains, livers, and blood were analyzed by gas chromatography--negative-ion-mass spectrometry for labeled fatty acids. To determine whether fatty acids were present in the brain apart from cerebral blood, a subset of animals was exsanguinated by perfusion with buffered saline, and the brain was then fractionated into subcellular components. Results demonstrated that the brain took up both labeled essential fatty acids within 8 h from the time of dosing. There was on average a greater uptake of linolenic acid into the cerebellum than into the cerebral cortex during the first 8 d of life in rats. The amount of linoleic acid taken into either region was similar, however. Docosahexaenoic acid intermediates, 20:5n-3 and 22:5n-3, were also found labeled in the brain. Time-course labeling experiments indicated that these intermediates may be converted to 22:6n-3 within the brain. A rise of labeled 22:6n-3 in the brain at 24 h appeared to be due to uptake of this fatty acid from the blood. The amount of labeled 22:6n-3 in the brain continued to increase beyond 24 h, and this did not appear to be correlated with its blood concentration. These results suggest that, during development in the rodent, different regions within the brain may vary in their capacity to synthesize 22:6n-3, and this may be correlated with regional growth rates. PMID- 8729104 TI - In vivo compartmental metabolism of 13C-docosahexaenoic acid, studied by gas chromatography-combustion isotope ratio mass spectrometry. AB - The exchange of docosahexaenoic acid (22:6n-3) within lipid pools in rat and human has been followed as a function of time after the ingestion of triglycerides (TG) containing 22:6n-3 labeled with 13C(13C 22:6n-3). The 13C abundance in the fatty acid was measured by gas-chromatography-combustion isotope ratio mass spectrometry which allowed the detection of 0.001 atom 13C percent 12C. The 13C 22:6n-3 appearance was rapid in the TG of very low density lipoprotein plus chylomicron fraction, in which the maximal labeling was observed at 3 and 2 h after ingestion in rat and human, respectively. Concomitant with the TG utilization of this fraction by lipoprotein lipase from tissues, unesterified 13C 22:6n-3 appeared in the plasma albumin. 13C 22:6n-3 bound to albumin was mostly present in unesterified form before 12 h post-ingestion while after that period, lysophosphatidylcholine (lysoPC) bound to albumin carried higher 13C 22:6n-3 concentrations. These lyso-PC were mostly from hepatic origin and might represent a potential source of 22:6n-3 redistribution to tissues. The 13C 22:6n 3 uptake into rat brain PC and phosphatidylethanolamine was still increasing when the concentration of plasma unesterified 13C 22:6n-3 had already dropped to a minimal plateau value and during the period of maximal plasma circulation of 13C 22:6n-3-lysoPC bound to albumin. In contrast, the uptake of 13C 22:6n-3 into blood platelet PC occurred during the phase of important circulation of 13C-22:6n 3 bound to albumin, suggesting the in vivo efficiency of the Lands pathway for this fatty acid. It is concluded that 13C 22:6n-3 esterified in TG is rapidly absorbed and redistributed within plasma lipoproteins and that its redistribution within the two lipid species bound to albumin might influence its uptake by platelets and rat brain. PMID- 8729106 TI - In vivo 13C nuclear magnetic resonance: applications and current limitations for noninvasive assessment of fatty acid status. AB - As a noninvasive method, in vivo 13C nuclear magnetic resonance has potentially important applications in understanding the metabolism of long chain fatty acids in organs of living humans. At present, this methodology is most advanced for research on glucose utilization. However, the main 13C signals visible in vivo are from fatty acids in adipose tissue and the olefinic signals can be used to noninvasively estimate adipose tissue content and relative dietary intake of polyunsaturates and monounsaturates. The low natural abundance of 13C improves the utility of this isotope for fatty acid tracer studies. Due to excessive signal broadening, uniform 13C-labelling seems to have limited application in in vivo fatty acid studies. Tracer fatty acids with 13C enrichment at a specific carbon position, i.e., [13-13C] gamma-linolenate, appear to be the most useful for in vivo tracer studies. Development of methods permitting resolution of 13C enrichment in structural lipids of lean tissues will be an important breakthrough which may make human tracer studies feasible and worthwhile. PMID- 8729105 TI - Synthesis of fat in response to alterations in diet: insights from new stable isotope methodologies. AB - Synthesis of fatty acids, or de novo lipogenesis (DNL), is an intensively researched metabolic pathway whose functional significance and metabolic role have nevertheless remained uncertain. Methodologic problems that limited previous investigations of DNL in vivo and recent methodologic advances that address these problems are discussed here. In particular, deuterated water incorporation and mass isotopomer distribution analysis techniques are described. Recent experimental results in humans based on these techniques are reviewed, emphasizing dietary and hormonal factors that modulate DNL and quantitative significance of DNL under various conditions, including carbohydrate overfeeding. The somewhat surprising finding that DNL appears not to be a quantitatively major pathway even under conditions of surplus carbohydrate energy intake, at least in normal adults on typical Western diets, is discussed in depth. Nutritional and metabolic implications of these results are also noted, and some speculations on possible functional roles of DNL in normal physiology and disease states are presented in this context. In summary, methodologic advances have added to our understanding of DNL and its regulation, but many questions concerning quantitation and function remain unanswered. PMID- 8729107 TI - Effect of dietary alpha-linolenic acid intake on incorporation of docosahexaenoic and arachidonic acids into plasma phospholipids of term infants. AB - The fractional conversion rates of plasma phospholipid alpha-linolenic acid (18:3n-3) and linoleic acid (18:2n-6) to docosahexaenoic acid (22:6n-3) and arachidonic acid (20:4n-6), respectively, and the fractional rates of incorporation of 22:6n-3 and 20:4n-6 into plasma phospholipids were determined in 27 healthy 3-wk-old term infants who had received formulas with approximately 16% of fat as 18:2n-6 and 0.4% (n = 6), 1.0% (n = 11), or 3.2% (n = 10) as 18:3n-3 from birth. The infants were given a single dose of both [U-13C] 18:2n-6 and [U 13C] 18:3n-3 with a feeding, and blood samples were collected 8, 12, and 24 h afterward for determination of the isotopic enrichments of the [M + 18] isotopomers of plasma phospholipid fatty acids by negative chemical ionization gas chromatography/mass spectrometry. A simple precursor/product compartmental model was used to estimate fractional rates of conversion and incorporation. All infants converted 18:3n-3 to 22:6n-3 and 18:2n-6 to 20:4n-6. Although the fractional rate of conversion of 18:3n-3 to 22:6n-3 did not differ among groups, the fractional rate of incorporation of 22:6n-3 into the plasma phospholipid fraction was greater in infants who received 3.2% vs. 0.4% or 1.0% 18:3n-3 (4.1 +/- 2.2 vs. 1.6 +/- 1.5 or 2.0 +/- 1.0% of the plasma phospholipid 22:6n-3 pool daily). The fractional rate of conversion of 18:2n-6 to 20:4n-6 was less in infants who received the 3.2% 18:3n-3 intake (0.4 +/- 0.3% of the plasma phospholipid 18:2n-6 pool daily vs. 1.1 +/- 0.7% and 0.8 +/- 0.5% in those who received 0.4 and 1.0% 18:3n-3, respectively). The fractional rate of incorporation of 20:4n-6 into plasma phospholipid also was less in the 3.2% vs. the 0.4 and 1.0% 18:3n-3 groups (2.7 +/- 1.4% vs. 5.9 +/- 2.6 and 4.4 +/- 1.7%, respectively, of the plasma phospholipid 20:4n-6 pool daily). PMID- 8729108 TI - Enzyme replacement and gene therapy for Gaucher's disease. AB - The lipid storage disorders have long been considered primary candidates for enzyme replacement therapy. This goal has been achieved with a remarkable degree of success in Gaucher's disease. Among the accomplishments that were important to obtain clinical benefit were the development of a large-scale procedure to purify human placental glucocerebrosidase and a method to target this enzyme to lipid storing macrophages through glycoform modification. In addition, the effectiveness of recombinantly produced macrophage-targeted glucocerebrosidase has recently been demonstrated. Because macrophages originate from stem cells in the bone marrow, ex vivo transduction of these cells with retroviral vectors containing the cDNA for human glucocerebrosidase is being explored for the genetic therapy of Gaucher's disease. PMID- 8729109 TI - Very long-chain fatty acids in diagnosis, pathogenesis, and therapy of peroxisomal disorders. AB - Abnormally high levels of very long-chain fatty acids (VLCFA) are a feature in nine of the fifteen peroxisomal disorders that have been identified so far. Saturated VLCFA accumulate in X-linked adrenoleukodystrophy, appear to disrupt membrane structure, and may play a role in the pathogenesis of a brain inflammatory response. Dietary therapy initiated when patients are still asymptomatic may be of clinical benefit. PMID- 8729110 TI - Docosahexaenoic acid therapy in docosahexaenoic acid-deficient patients with disorders of peroxisomal biogenesis. AB - A patient with classic Zellweger syndrome was treated with docosahexaenoic acid ethyl ester (DHA-EE) for three months. Five other patients with Zellweger variants (four of them less than one-year-old and a five-year-old) were treated with DHA-EE until normalization of the DHA levels in erythrocytes. When arachidonic acid (AA) concentration decreased, AA was added to the diet. Thereafter, a combined treatment with DHA plus AA followed, in a variable proportion that allowed the high levels of DHA in erythrocytes to be maintained. In the patient with Zellweger syndrome, DHA therapy produced an increase in plasmalogen and a decrease in 26:0 and 26:1. No clear clinical improvement could be detected in this patient during the short period of treatment with DHA-EE. The most consistent clinical effect produced by DHA therapy in the other patients with disorders of peroxisomal biogenesis was visual improvement, even in those patients that were virtually blind before the treatment. In general, the developmental curve began to accelerate. The infants became more alert, acquired better visual and social contact and muscular tone improved, with the beginning of good head control. The liver tests tended to normalize and some patients showed a reduction of hepatomegaly. All these favorable changes occurred when the patients were taking the DHA-EE alone. In some of the patients, muscular tone seemed to improve further after introducing AA supplements. From the biochemical point of view, the plasmalogen levels increased in most cases in erythrocytes, and the two ratios 26:0/22:0 and 26:1/22:0 decreased in plasma. In some patients there was a tendency for 26:1 to increase in plasma and for 18:0 plasmalogen to decrease in erythrocytes when AA was introduced in the diet. The significance of these findings remains to be elucidated, but they stress the importance of strict monitoring and control of the polyunsaturated fatty acids status during DHA therapy. PMID- 8729111 TI - Losses of arachidonic acid in rat liver after alcohol inhalation. AB - This paper presents an animal model of alcoholism in which rats were exposed to alcohol by inhalation and were fed a diet that simulated the poor diet of some alcoholics. It is hypothesized that some of the pathophysiological effects of alcohol are related to its effects on essential fatty acid metabolism and composition of vital organs. A diet that contains no 20- and 22-carbon essential fatty acids and has low levels of 18-carbon essential fatty acids was used as a dietary challenge. Addition of a second metabolic challenge, i.e., alcohol, led to loss of tissue polyunsaturates, particularly liver arachidonate. A method of cycling alcohol inhalation for 12 h/d was also presented, which was also shown to lower liver arachidonic acid content. PMID- 8729112 TI - Arachidonic acid to eicosapentaenoic acid ratio in blood correlates positively with clinical symptoms of depression. AB - In this study of 20 moderately to severely depressed patients, diagnosed using current research diagnostic criteria and excluding known bipolar affective disorder and reactive depression, we investigated relationships between severity of depression and levels and ratios of n-3 and n-6 long-chain polyunsaturated fatty acids (PUFA) in plasma and erythrocyte phospholipids (PL). Severity of depression was measured using the 21-item Hamilton depression rating scale (HRS) and a second linear rating scale (LRS) of severity of depressive symptoms that omitted anxiety symptoms. There was a significant correlation between the ratio of erythrocyte PL arachidonic acid (AA) to eicosapentaenoic acid (EPA) and severity of depression as rated by the HRS (P < 0.05) and the LRS for depression (P < 0.01). There was also a significant negative correlation between erythrocyte EPA and the LRS (P < 0.05). The AA/EPA ratio in plasma PL and the ratio of erythrocyte long-chain (C20 and C22 carbon) n-6 to long-chain n-3 PUFA were also significantly correlated with the LRS (P < 0.05). These findings do not appear to be simply explained by differences in dietary intake of EPA. We cannot determine whether the high ratios of AA/EPA in both plasma and erythrocyte PL are the result of depression or whether tissue PUFA change predate the depressive symptoms. We suggest, however, that our findings provide a basis for studying the effect of the nutritional supplementation of depressed subjects, aimed at reducing the AA/EPA ratio in tissues and severity of depression. PMID- 8729113 TI - Fatty acids and schizophrenia. AB - In a controlled study of red cell membrane fatty acids in patients with schizophrenia, substantial depletions of fatty acids from both the n-6 and n-3 series were demonstrated. Arachidonic acid and docosahexaenoic acid were particularly depleted. In a separate study, dietary analysis revealed no deficiency of fatty acid intake in this patient group, but greater intake of n-3 fatty acids was associated with less severe symptomatology. Dietary supplementation for six weeks with 10 g per day of concentrated fish oil (MaxEPA) led to significant improvement in schizophrenic symptoms. This clinical improvement was related to the increased level of n-3 fatty acids in red cell membranes. These findings form part of a growing body of research data suggestive of an abnormality in cell membrane fatty acid composition in schizophrenia. The preliminary evidence for clinically effective dietary manipulation to correct such an abnormality opens up novel and exciting therapeutic possibilities. PMID- 8729115 TI - Effect of dietary docosahexaenoic acid on brain composition and neural function in term infants. AB - There is a need to determine whether there is a dietary requirement for docosahexaenoic acid (DHA, 22:6n-3) by term infants to achieve their full developmental potential. Studies of brain fatty acid composition demonstrated that infants who were breast fed had greater levels of cerebral cortex DHA than did infants who were formula fed, suggesting that DHA in the cerebrum is dependent on a supply in the diet. Some physiological studies reported that electrophysiological and behavioral assessments of visual function were improved in breast-fed infants relative to those fed formula and that this was related to the length of breast feeding. While some randomized studies of DHA supplementation of infant formula to term infants demonstrated that the visual function of formula-fed infants could be improved to breast-fed levels by adding DHA to formula, others failed to demonstrate an effect. Variations in dietary treatments and methods of assessment make comparison of the studies difficult. Further work is necessary to rigorously establish if there are long-term benefits of dietary DHA to the term infant. PMID- 8729116 TI - Increased docosahexaenoic acid levels in human newborn infants by administration of sardines and fish oil during pregnancy. AB - In rhesus monkeys, maternal n-3 fatty acid deficiency during pregnancy produces infant monkeys deficient in n-3 fatty acids at birth. These results stimulated current experiments to find out if n-3 fatty acids from fish in the diets of pregnant women would influence the concentration of docosahexaenoic acid (DHA, 22:6 n-3) in the newborn human infant. Fifteen healthy pregnant women were enrolled to receive a 9-wk dietary supplementation of n-3 fatty acids from the 26th to the 35th wk of pregnancy. Sixteen pregnant women were not supplemented and served as controls. n-3 Fatty acid supplementation consisted of sardines and additional fish oil, which provided a total of 2.6 g of n-3 fatty acids per day (d) for the 9-wk period of supplementation. This included 1.01 g DHA. The end point of this study was the blood concentrations of DHA in the newborn infant. DHA in maternal red blood cells increased from 4.6% of total fatty acids to 7.15% at the end of the supplement period and at the time of delivery decreased (as expected) to 5.97% of total fatty acids. Maternal plasma showed a similar change from 2.12 to 3.51% of total fatty acids and then decreased to 2.35%. Levels of DHA in plasma and red blood cells of unsupplemented mothers did not change during the same time period. Levels of DHA in blood of newborn infants differed greatly in infants born from n-3-supplemented mothers compared with control infants. In red blood cells, DHA was 7.92% of total fatty acids compared with 5.86% (control infants). Plasma values showed a similar difference: 5.05% vs. 3.47% (controls). In n-3-supplemented infants, DHA concentrations were 35.2% higher than in control infants in red blood cells and 45.5% higher in plasma. These data indicate the importance of maternal dietary n-3 fatty acids and, in particular, maternal dietary DHA in promoting higher concentrations of DHA in the blood of the newborn infant. PMID- 8729114 TI - Role of essential fatty acids in the function of the developing nervous system. AB - The basis for n-3 fatty acid essentially in humans includes not only biochemical evidence but functional measures associated with n-3 deficiency in human and nonhuman primates. Functional development of the retina and the occipital cortex are affected by alpha-linolenic acid deficiency and by a lack of docosahexaenoic acid (DHA) in preterm infant formulas and, as reported more recently, in term diets. Functional effects of n-3 supply on sleep-wake cycles and heart rate rhythms support the need for dietary n-3 fatty acids during early development. Our results indicate that n-3 long-chain polyunsaturated fatty acids should be considered provisionally essential for infant nutrition. DHA may also be required by individuals with inherited metabolic defects in elongation and desaturation activity, such as patients with peroxisomal disorders and some forms of retinitis pigmentosa. PMID- 8729117 TI - Polyunsaturation in cell membranes and lipid bilayers and its effects on membrane proteins. AB - The effect of variation of the degree of cis-unsaturation on cell membrane protein functioning was investigated using a model lipid bilayer system and protein kinase C (PKC). This protein is a key element of signal transduction. Furthermore it is representative of a class of extrinsic membrane proteins that show lipid dependent interactions with cell membranes. To test for dependence of activity on the phospholipid unsaturation, experiments were devised using a vesicle assay system consisting of phosphatidylcholine (PC) and phosphatidylserine (PS) in which the unsaturation was systematically varied. Highly purified PKC alpha and epsilon were obtained using the baculovirus-insect cell expression system. It was shown that increased PC unsaturation elevated the activity of PKC alpha. By contrast, increasing the unsaturation of PS decreased the activity of PKC alpha, and to a lesser extent PKC epsilon. This result immediately rules out any single lipid bilayer physical parameter, such as lipid order, underlying the effect. It is proposed that while PC unsaturation effects are explainable on the basis of a contribution to membrane surface curvature stress, the effects of PS unsaturation may be due to specific protein-lipid interactions. Overall, the results indicate that altered phospholipid unsaturation in cell membranes that occurs in certain disease states such as chronic alcoholism, or by dietary manipulations, are likely to have profound effects on signal transduction pathways involving PKC and similar proteins. PMID- 8729118 TI - A role for phospholipid polyunsaturation in modulating membrane protein function. AB - Visual transduction is one of the best characterized G protein--coupled signalling systems. In addition, about 50% of the disk membrane phospholipid acyl chains are 22:6n-3, making this system ideal for determining the role of polyunsaturation in modulating membrane-signalling systems. The extent of formation of metarhodopsin II (MII), the G protein--activating photointermediate of rhodopsin, was studied in phospholipid vesicles composed of a variety of phosphatidylcholines, differing in their acyl chain composition at the sn-2 position. The amount of MII formed increased progressively with the level of acyl chain unsaturation at the sn-2 position. The effect of added cholesterol was to reduce the amount of MII formed. The acyl chain packing free volume of the rhodopsin containing lipid vesicles was characterized by a fractional volume parameter fv derived from measurements of the time-resolved fluorescence anisotropy decay of the hydrophobic membrane probe 1,6-diphenyl-1,3,5-hexatriene. The relationship among sn-2 acyl chain unsaturation, cholesterol content, and MII formation is explained on the basis of variation in fv with bilayer lipid composition and a novel model for the packing of phospholipids containing polyenoic acyl chains, such as 22:6n-3. PMID- 8729119 TI - Nuclear magnetic resonance investigation of hydrocarbon chain packing in bilayers of polyunsaturated phospholipids. AB - 2H nuclear magnetic resonance (NMR) on chain-deuterated phospholipids has been used to study the influence of the degree of unsaturation on lipid chain packing and on area per molecule at the lipid water interface. Order and motions of deuterated stearic acid in position sn-1 of phosphatidylcholines (PC) containing 18:0, 18:1n-9, 18:2n-6, 18:3n-3, 20:4n-6, 20:5n-3, or 22:6n-3 in position sn-2 were investigated in pure PC and in mixtures of PC in a phosphatidylethanolamine (PE) matrix. Results reveal that lipid packing in bilayers is mainly controlled by packing requirements at the lipid water interface. Increasing degrees of unsaturation lower chain order and increase area per PC molecule, whereas inclusion of PE in model membranes has the opposite effect. Chain order and motions in highly unsaturated lipid membranes are less sensitive to changes in temperature. Temperature sensitivity decreases further upon incorporation of PC into a PE matrix. Unsaturation induces chain disordering, which may be interpreted as an increase in area per molecule of lipids toward the center of the bilayer. This may result in a lower packing density of unsaturated lipids at the lipid water interface. We hypothesize that these differences in lipid packing and dynamics may influence activity of membrane proteins. PMID- 8729120 TI - Membrane docosahexaenoic acid vs. eicosapentaenoic acid and the beating function of the cardiomyocyte and its regulation through the adrenergic receptors. AB - The beta-adrenergic system in cardiac muscle cells is influenced by the n-3 polyunsaturated fatty acid (PUFA) content in cell membranes. This study deals with the specific effect of docosahexaenoic acid (DHA) as compared to other n-3 PUFA, without modification of the arachidonic acid content. Increasing the DHA content in the phospholipids of isolated cardiomyocytes did not provoke electrical or contraction modifications, except for a slightly lower plateau phase (-2 mV). Conversely, the beta-adrenergic function was affected at several levels: (i) the receptor affinity for dihydroalprenolol tended to decrease (Kd) without alteration of the number of beta-binding sites (beta max); (ii) the isoproterenol-induced (10(-7) M and 10(-6) M) cAMP production was significantly decreased (-20%); and (iii) the positive chronotropic response to beta-adrenergic stimulation (isoproterenol, 10(-7) M) was significantly enhanced (+80%). In order to further investigate the relationship between the decreased cAMP and the increased chronotropic response, the cells were treated with dibutyryl-cAMP, a permeant analogue of cAMP, which elicited a significantly higher chronotropic response in DHA-enriched cells than in EPA-enriched cells +50%). The increase in DHA content in cardiac cell membranes phospholipids may thus affect the beta adrenergic system through an increase of cAMP efficiency. Although the membrane phosphatidylinositols were largely involved in the PUFA alterations, none of the parameters related to alpha-adrenergic system (chronotropic response, receptor density, affinity for prasozin, and inositide phosphate production) were influenced by the membrane DHA content. PMID- 8729121 TI - Marked enrichment of the alkenylacyl subclass of plasma ethanolamine glycerophospholipid with eicosapentaenoic acid in human subjects consuming a fish oil concentrate. AB - Alteration in human platelet fatty acid levels with the consumption of fish oils containing eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3) have been well documented, but changes in the fatty acid composition of plasma plasmalogenic phospholipid under similar circumstances have not been delineated. In the present study, subjects consumed the fish oil concentrate (MaxEPA) for 6 wk followed immediately by a 6-wk recovery period with no fish oil ingestion. Plasma total choline glycerophospholipid (GPC) and ethanolamine glycerophospholipid (GPE) subclasses isolated from blood samples obtained at 0, 3, 6, 9 and 12 wk of the experimental period were analyzed for fatty acid composition via thin-layer and gas-liquid chromatographic techniques. Consumption of fish oil for 3 or 6 wk significantly elevated the content of n-3 fatty acids while concomitantly decreasing n-6 fatty acid levels in plasma total GPC and in diacyl and alkenylacyl (plasmalogen) GPE. Alkenylacyl GPE exhibited the greatest alteration of both n-3 and n-6 fatty acid levels. Following 6 wk of supplementation with fish oil, EPA rose by 24.6 mol% in alkenylacyl GPE compared to increases of 6.7 and 7.1 mol% in diacyl GPE and total GPC, respectively. The increase in EPA (from 5.0 to 29.6 mol%) in plasma alkenylacyl GPE represents amongst the highest enrichment of EPA in any lipid yet reported in human subjects. DHA also rose by 8.0, 4.8, and 3.1 mol% in alkenylacyl GPE, diacyl GPE, and total GPC, respectively. Alkenylacyl GPE exhibited the greatest mol% decline (by 18.7 mol%) in arachidonic acid (AA, 20:4n-6) following 6 wk of fish oil supplementation. The corresponding decreases of AA in diacyl GPE and total GPC were 8.7 and 1.8 mol%, respectively. Following the 6 wk recovery period, n-3 and n-6 fatty acid levels had returned to pre-supplementation values. The marked enrichment of alkenylacyl GPE in n-3 fatty acids, especially EPA, may be of significance with respect to a unique role for this plasma phospholipid subclass in attenuating certain lipoprotein-mediated cardiovascular effects as observed with fish/fish oil consumption. PMID- 8729122 TI - Thioesterification of platelet proteins with saturated and polyunsaturated fatty acids. AB - We have demonstrated that more than 20 platelet proteins can be acylated with fatty acids via thioester linkages. These include the glycoprotein IX beta chain of glycoprotein Ib, components of the von Willebrand factor receptor on the platelet surface, P-selectin, and alpha subunits of Gz, Gq, and Gi. Our studies have shown that platelet proteins can be posttranslationally acylated in thioester linkages not only with palmitate but with myristate and also with the eicosanoid precursor fatty acids arachidonate and eicosapentaenoate. Thioesterification of platelet proteins with fatty acids other than palmitate may have significant functional consequences for reversible binding of proteins to membranes. PMID- 8729123 TI - Membrane-associated and cytoplasmic fatty acid-binding proteins. AB - A number of cellular fatty acid-binding proteins are being implicated in the uptake and intracellular transport of long-chain fatty acids by parenchymal cells. Having been a topic of research for more than 20 years, cytoplasmic fatty acid-binding proteins now are assigned various pivotal functions in intracellular fatty acid transport and metabolism. More recently several membrane-associated fatty acid-binding proteins have been identified and these proteins are thought to function in the transmembrane transport of fatty acids. In this review, a short summary is provided of the latest developments in this research area. PMID- 8729124 TI - Specificity of polyunsaturated fatty acid release from rat brain synaptosomes. AB - Release of specific polyunsaturated fatty acids for cell membranes may have a significant implication in biological function, considering the involvement of various fatty acids in cell signal transduction. In the present study, release of polyunsaturated fatty acids from rat brain synaptosomes by endogenous synaptosomal lipase activity was examined in comparison to that by cobra venom phospholipase A2 (Naja naja naja). Cobra venom phospholipase A2 (Naja naja naja) preferentially hydrolyzed docosahexaenoic acid (22:6n-3) from both synaptosomes and lipid mixtures containing similar classes of lipids commonly found in the brain. Arachidonic acid (20:4n-6) and oleic acid (18:1n-9) were also hydrolyzed; however, monoene species was hydrolyzed slower than were polyenoic species in synaptosomes. Phosphatidylethanolamine was the most preferred phospholipid class for release of 22:6n-3 fatty acid from both lipid mixtures and synaptosomes. In contrast to hydrolysis by cobra venom phospholipase A2, endogenous synaptosomal lipase activity preferentially hydrolyzed 20:4n-6 from rat brain synaptosomes, despite the high abundance of 22:6n-3 in synaptosomal membranes. Preferential release of 20:4n-6 was observed over a wide range of pH values and calcium concentrations. Synaptosomal 22:6 species appeared to be resistant to hydrolysis even after stimulation with various agents such as phorbolmyristate, suggesting that physiological importance of 22:6n-3 in neuronal membranes may not be as the release fatty acid. PMID- 8729125 TI - Natural and accelerated docosahexaenoic acid accumulation in the prenatal rat brain. AB - The quantity and distribution of docosahexaenoic acid (DHA) in major brain phospholipids (PL) was examined in the fetal rat brain before birth, using thin layer and capillary column gas chromatography. A rapid increment of DHA content of about 187 micrograms/g brain/day was observed between 17 to 20 days gestation, as opposed to 39.3 +/- 2.9 micrograms/g brain/day prior to that. Single intraamniotic injections of 5 microL ethyl-docosahexaenoate (Et-DHA) (12 microM, 4.25 mg) administered to 17-day-old fetuses were used to examine the uptake of DHA into brain PL. Three days following injection, the amount of n-3 polyunsaturated fatty acids increased by 28% compared to ethyl-oleate (Et-Ole) injected fetuses. Compared to the n-6 fatty acid family, the relative amount of DHA increased in the phosphatidylserine (PS), phosphatidylethanolamine and phosphatidylcholine (PC) lipids by 15 (P = 0.02), 13 and 14%, respectively. A major increase in the pool size of phosphatidylinositol and PS (110 and 50.3%, respectively), and a decrease in PC (8.2%) were observed 3 d after Et-DHA as compared to Et-Ole administration. The data suggest that a single intraamniotic administration of Et-DHA can modulate membrane PL content and alter PUFA composition. PMID- 8729126 TI - n-3 polyunsaturated fatty acids and human cytokine synthesis. AB - The cytokines interleukin-1 and tumor necrosis factor-alpha are potent peptide mediators of the immune response. They exert potent cellular effects at nanomolar concentrations with target tissues ranging from the vascular endothelium, to the central nervous system and immunocompetent cells. The synthesis of these two cytokines can be suppressed pharmacologically by steroids and other immunosuppressive agents such as cyclosporin A. In previous studies we have found that such an inhibition of inflammatory cytokine synthesis can also be achieved by increasing the content of n-3 polyunsaturated fatty acids in leukocyte membranes. PMID- 8729127 TI - Effects of modulation of inflammatory and immune parameters in patients with rheumatic and inflammatory disease receiving dietary supplementation of n-3 and n 6 fatty acids. AB - To describe the rationale and status of n-3 and n-6 fatty acid dietary supplementation in patients with inflammatory disease. The most recent literature is reviewed with a focus on rheumatoid arthritis (RA) as most investigations have described the use of n-3 supplements in this disease entity. Investigations from Europe, the United States, and Australia have described consistent improvement in tender joint scores with many investigators also observing improvements in morning stiffness. A meta analysis has confirmed the predictable improvement in tender joints. Recent studies also suggest that some patients with RA are able to discontinue nonsteroidal antiinflammatory drugs (NSAIDs) while receiving n-3 fatty acids. A large number of peer reviewed publications from around the world have established the utility of dietary supplementation with n-3 fatty acids in reducing tender joint counts and morning stiffness in patients with RA. Some patients are also able to discontinue NSAIDs while on these supplements. PMID- 8729128 TI - The significance of polyunsaturated fatty acids in cutaneous biology. AB - The skin epidermis displays a highly active metabolism of polyunsaturated fatty acids (PUFA). Dietary deficiency of linoleic acid (LA) and 18-carbon (n-6) PUFA results in characteristic scaly skin disorder and excessive epidermal water loss. Arachidonic acid, a 20-carbon (n-6) PUFA is metabolized via the cyclooxygenase pathway into predominantly prostaglandin E2 (PGE2) PGF2 alpha, and PGD2 and via the lipoxygenase pathway into predominantly 15-hydroxyeicosatetraenoic acid (15 HETE). The prostaglandins modulate normal skin physiological processes at low concentrations and inflammatory reactions at high concentrations. Similarly, the very active epidermal 15-lipoxygenase transforms dihomogammalinolenic acid (DGLA) into 15-hydroxy eicosatrienoic acid (15-HETrE), eicosapentaenoic acid (EPA) into 15-hydroxyeicosapentaenoic acid (15-HEPE) and docosahexaenoic acid (DHA) into 17 hydroxydocosahexaenoic acid (17-HDoHE), respectively. These monohydroxy acids exhibit anti-inflammatory properties. In contrast, the 18-carbon (n-6) PUFA is transformed into 13-hydroxy-9,11-octadecadienoic acid (13-HODE), which exerts antiproliferative properties in the tissue. Thus, the supplementation of diets with appropriate purified vegetable oils and/or fish oil may generate local cutaneous anti-inflammatory metabolites which could serve as a less toxic in vivo monotherapy or as adjuncts to standard therapeutic regimens for the management of skin inflammatory disorders. PMID- 8729129 TI - Quantification of distinct molecular species of platelet activating factor in ulcerative colitis. AB - The aim of this study was to characterize the synthesis and metabolism of platelet activating factor (PAF, 1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine) by colonic mucosa from patients with ulcerative colitis and healthy individuals. Tissue was obtained by endoscopic biopsy and by scraping the mucosa from surgical resections. Tissue was assayed for the various molecular species of PAF and its biologically inactive metabolite lyso-PAF using gas chromatography/mass spectrometry. Mucosa from surgical resections for ulcerative colitis contained C16:0 PAF (mean = 156 ng/g of mucosa), but not C18:0 PAF, PAF was not identified in mucosa from normal surgical resections or in endoscopic biopsies from either patients with ulcerative colitis or normal individuals. Both C16:0 lyso-PAF and C18:0 lyso-PAF were found in mucosa from normal and ulcerative colitis surgical resections and in endoscopic biopsies from ulcerative colitis and normal tissue. Levels of lyso-PAF were similar in ulcerative colitis and normal mucosa. Incubation of mucosa from areas of active inflammation in ulcerative colitis with the calcium ionophore A23187 increased the levels of C16:0 PAF by 2-3 fold and also increased the levels of C16:0 lyso-PAF. Addition of 3H-PAF to endoscopic biopsies from either normal individuals or patients with ulcerative colitis resulted in hydrolysis to 3H-lyso-PAF. The data on colonic mucosal levels of PAF are consistent with the results of earlier studies measuring PAF in patients with ulcerative colitis by bioassay. This study examines the synthesis and metabolism of specific molecular species of PAF in ulcerative colitis for the first time. PMID- 8729130 TI - Skeletal muscle membrane lipids and insulin resistance. AB - Skeletal muscle plays a major role in insulin-stimulated glucose disposal. This paper reviews the range of evidence in humans and experimental animals demonstrating close associations between insulin action and two major aspects of muscle morphology: fatty acid composition of the major structural lipid (phospholipid) in muscle cell membranes and relative proportions of major muscle fiber types. Work in vitro and in vivo in both rats and humans has shown that incorporation of more unsaturated fatty acids into muscle membrane phospholipid is associated with improved insulin action. As the corollary, a higher proportion of saturated fats is linked to impairment of insulin action (insulin resistance). Studies in vitro suggest a causal relationship. Among polyunsaturated fatty acids (PUFA) there is some, but not conclusive, evidence that omega-3 (n-3) PUFA may play a particular role in improving insulin action; certainly a high n-6/n-3 ratio appears deleterious. In relation to fiber type, the more highly oxidative, insulin-sensitive type 1 and type 2a fibers have a higher percentage of unsaturated fatty acids, particularly n-3, in their membrane phospholipid, compared to the insulin-resistant, glycolytic, type 2b fibers. These variables, however, can be separated and may act in synergy to modulate insulin action. It remains to establish whether lifestyle (e.g., dietary fatty acid profile and physical activity), genetic predisposition, or a combination are the prime determinants of muscle morphology (particularly membrane lipid profile) and hence insulin action. PMID- 8729131 TI - Role of lipids in development of noninsulin-dependent diabetes mellitus: lessons learned from Pima Indians. AB - We studied the role of lipids in the pathogenesis of noninsulin-dependent diabetes mellitus (NIDDM) in Pima Indians. High plasma levels of nonesterified fatty acid (NEFA) predicted development of NIDDM, but this effect cannot entirely be explained by the glucose-fatty acid cycle. Dyslipidemia, although often associated with diabetes, did not seem to predict NIDDM and might rather be associated with, or the consequence of insulin resistance. In some individuals, a single amino acid substitution in the intestinal fatty acid binding protein could result in increased rates of intestinal absorption of dietary NEFA and thereby contribute to increased lipid-oxidation rates and insulin resistance. PMID- 8729132 TI - Arachidonic acid maldistribution in obesity. AB - Arachidonic acid is an important regulator of cellular function via its effects on the physical properties of membranes, in its free form, or as a substrate for eicosanoids. Dietary studies indicate that its production is regulated, but the mechanisms of this regulation and factors influencing arachidonate distribution from the site of production remain to be determined. In particular, whether there is a nonoxidative fate for arachidonate once it has been released from phospholipid has yet to be determined. Variations in the arachidonate content of serum, liver, and muscle lipid fractions have been correlated with alterations in lipogenesis and insulin action, implying a role for arachidonate in fuel partitioning. Evidence for this mechanism acting systemically has been found in genetic models of obesity in rodents and also in humans. This review proposes that variation in the distribution of arachidonate between phospholipid and cholesteryl ester fractions participates in the abnormal fuel partitioning associated with some forms of genetic obesity. PMID- 8729133 TI - Gender differences in insulin-induced free fatty acid suppression: studies in an African American population. AB - This study of African Americans (AA) was designed to investigate gender differences in insulin-induced free fatty acid (FFA) suppression. Sixty AA (34 women, mean age 34 +/- 7.6 years, and 26 men, mean age 30 +/- 2.9 years) participated. All subjects had an oral glucose tolerance test (OGTT). Nineteen women and 18 men also underwent a euglycemic hyperinsulinemic clamp (IC) study. Plasma insulin and FFA concentrations were obtained during both tests at 0, 60, and 120 min. While there was no gender difference in body mass index (P = 0.21), women had greater percent body fat (P < 0.001) calculated by the Siri formula. There was no gender difference in fasting FFA levels, but during the OGTT, women compared to men had significantly greater FFA suppression. Both nonobese and obese women suppressed FFA concentration by 88%, and nonobese and obese men suppressed FFA concentration by 80 and 66% respectively. This gender difference in FFA suppression was significant (P = 0.001) and independent of obesity and insulin concentration. During the IC studies, there were no gender or obesity differences in FFA suppression, with women and men suppressing FFA levels by 87 89% (P = 0.7). Fasting insulin concentrations were higher in obese vs. nonobese (P = 0.03), but fasting FFA concentrations were not different (P = 0.15). For nonobese and obese females, fasting FFA levels were 0.55 +/- 0.24 and 0.44 +/- 0.26 mEq/L, respectively, and for nonobese and obese males, 0.45 +/- 0.2 and 0.35 +/- 0.18 mEq/L, respectively. In women, development of obesity may be enhanced by greater sensitivity to insulin-induced FFA suppression as measured during an OGTT. To detect gender differences in FFA metabolism, the OGTT is superior to the IC. The lack of elevation in fasting FFA levels in obese AA women and men has not been reported in other racial groups and may indicate a greater adipocyte sensitivity to insulin in AA. PMID- 8729134 TI - Trans, n-3, and n-6 fatty acids in Canadian human milk. AB - The presence of trans fatty acids in human milk may be a concern because of their possible adverse nutritional and physiological effects on the recipient infant. The mother's diet is the source of human milk trans fatty acids, and since these fatty acids are prevalent in many common foods of the Canadian diet, the trans fatty acid content and the fatty acid composition of Canadian human milk were measured by gas-liquid chromatography coupled with silver nitrate-thin layer chromatography. In samples obtained from 198 lactating mothers across Canada, the average percentage of total trans (sum of t18:1, t18:2, and t18:3) was 7.2% of breast milk fatty acids with a range of 0.1-17.2%. Analysis of t18:1 isomer distribution indicated that partially hydrogenated vegetable oils are the major source of these trans fatty acids in human milk, whereas contribution from dairy products appeared to be relatively minor. Linoleic and alpha-linolenic acid levels were inversely related to the total trans fatty acids, indicating that the elevation of trans fatty acids in Canadian human milk is at the expense of n-3 and n-6 essential fatty acids. Levels of arachidonic and docosahexaenoic acids did not correlate with their parent fatty acids, indicating that it might be difficult to elevate the levels of n-6 and n-3 c20-22 polyunsaturated fatty acids in breast milk by increasing levels of linoleic and alpha-linolenic acids in the mother's diet. PMID- 8729135 TI - Characteristics of fats in Japanese diets and current recommendations. AB - Although there is no firm evidence to support the "ideal" or even "appropriate" healthy level of dietary fat, the habitual fat consumption pattern in Japan seems to be a criterion for the recommended allowance both in the quantitative and qualitative points of view as judged from the life expectancy and the incidence of degenerative diseases. The new recommended dietary allowance of Japan, fifth revision effective for five years starting in 1995, adopted dietary fat levels of 20-25 energy percent, the ratio of saturated, monounsaturated, and polyunsaturated fatty acids at 1:1.5:1 and the ratio of n-6/n-3 at 4. The recommended fat level is similar to that previously consumed in Japan, and is even lower than that in diets used to treat hyperlipidemia in Western countries, current recommendations in those countries being 30 energy percent fat. Convincing data for the beneficial effects of n-3 polyunsaturated fatty acids on human health, in particular for healthy people, have been presented in only a few reports. However, the recommended n-6/n-3 ratio of 4 seems reasonable compared with the ratio of around 10 in other developed countries. In this context, it is more important to fully understand the nutritional and physiological roles of fat in healthy people rather than in those with chronic disease. At present, the low fat dietary pattern in Japan appears to be a healthy way of eating. PMID- 8729136 TI - Fats in Indian diets and their nutritional and health implications. AB - To arrive at rat requirements for Indians, the contribution of invisible fat should be determined. Total lipids were extracted from common Indian foods, and their fatty acid compositions were determined. This data and information on intake of various foods were used to estimate the contents of "invisible" fat and fatty acids in Indian diets. Taking into account World Health Organization (WHO) guidelines and the invisible-fat intake of Indians, recommendations were made for lower and upper limits of visible fats. In the rural poor, the "visible"-fat intakes are much lower than estimated minimum requirements. Therefore, to meet the energy needs of low-income groups, particularly young children, visible-fat intakes must be increased to recommended levels. The urban high-income group, however, should reduce dietary fat. Data on intake of various fatty acids in total diet shows that even the recommended lower limit of oil can meet linoleic acid requirements. Intake of alpha-linolenic acid is low, however. Increase in dietary n-3 polyunsaturated fatty acid (PUFA) produces hypolipidemic, anti inflammatory, and antithrombotic effects. Effects of n-3 PUFA on blood lipids, platelet fatty acid composition, and platelet aggregation were therefore investigated in Indian subjects consuming cereal-based diets. Supplementation of fish oils (long-chain n-3 PUFA) as well as the use of rapeseed oil (alpha linolenic acid) produced beneficial effects. Since the requirements of alpha linolenic acid and/ or long-chain n-3 PUFA are related to linoleic acid intake, use of more than one oil (correct choice) is recommended for providing a balanced intake of various fatty acids. Analysis of Indian food showed that some foods are good sources of alpha-linolenic acid. Regular consumption of these foods can also improve the quality of fat in Indian diets. Nonvegetarians, however, have the choice of eating fish to accomplish this. PMID- 8729138 TI - Production of eicosapentaenoic acid from marine bacteria. AB - A marine bacterium, judged as a new species close to Shewanella putrefaciens, was isolated from the intestinal contents of the Pacific mackerel. The isolated strain SCRC-2378 produced eicosapentaenoic acid (EPA) as the sole polyunsaturated fatty acid, which amounted to 24-40% of the total fatty acid in the cell, which corresponded to 2% of dry cell weight. Under the optimal growth conditions (pH 7.0, 20 degrees C, and grown aerobically for 12-18 h), the yield of SCRC-2738 reached 15 g of dry cells/L or 2 x 10(10) viable cells/mL. EPA existed as phospholipid and was found in the sn-2 position of phosphatidylethanolamine and phosphatidylglycerol. The 38 kbp (1,000 base pairs) genome DNA fragment was cloned from SCRC-2738 and expressed in Escherichia coli, which resulted in the production of EPA. The nucleotide sequence of the 38 kbp DNA fragment was determined. The DNA fragment contains eight open reading frames, and three of them possess homology with enzymes involved in fatty acid synthesis. Thus, it may be possible that these EPA biosynthesis genes are applied for EPA production in yeasts or higher plants, and offer a new method for EPA synthesis as new foods containing EPA. PMID- 8729137 TI - Impact of fat substitutes on fat intake. AB - Dietary fat is the number one nutrition concern of Americans. In response to rising consumer demand for reduced-fat foods, the food industry has developed a multitude of nonfat, lowfat, and reduced-fat versions of regular food products. To generate reduced-fat or fat-free products that have the same organoleptic characteristics of the regular fat version, food manufacturers frequently employ fat substitutes in the formulation of these foods. Fat substitutes are made from either carbohydrate, protein, or fat, or a combination of these components. Researchers have questioned the impact of fat substitutes on both fat and caloric intake. The majority of research studies in which fat substitutes were either covertly or overtly substituted for dietary fat indicate that in short-term, carefully-controlled conditions, fat substitutes can decrease both dietary fat intake and percentage of calorie intake from fat. However, individuals compensate for the caloric deficit created by the fat substitutes by increasing their consumption of other macronutrients, primarily carbohydrate. The long-term effect of fat substitutes on the fat intake of free-living individuals and weight control are unknown. People tend to eat more of a food when they know that food is reduced in fat. Fat substitutes should not be considered a substitute for sound nutrition education and a healthy lifestyle which includes regular exercise. PMID- 8729139 TI - Eicosapentaenoic acid suppressed the proliferation of vascular smooth muscle cells through modulation of various steps of growth signals. AB - Among fatty acids, only n-3 polyunsaturated fatty acids such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) inhibited the proliferation of vascular smooth muscle cells. DHA was less effective than EPA. To clarify the anti proliferative effect of n-3 polyunsaturated fatty acid, we have explored the effect of EPA on the signal transduction pathway of platelet derived growth factor (PDGF). EPA inhibited PDGF binding on its receptor and activation of protein kinase C. EPA also suppressed c-fos mRNA expression, one of immediate early genes, through partly inhibiting c-fos transcription. These data revealed that EPA could inhibit vascular smooth muscle cell proliferation through modulating various steps of the signal transduction by PDGF. PMID- 8729140 TI - The structure-activity relationship of lipoxygenase products of long-chain polyunsaturated fatty acids: effects on human platelet aggregation. AB - The effect of hydroperoxy and hydroxy derivatives of various fatty acids on human platelet aggregation was determined to delineate potencies and structure-activity function. In this regard, the 22-carbon n-3 fatty acids are the most potent inhibitors in comparison to the n-6 lipoxygenase derivatives. Submicromolar levels of the docosapentaenoic (22:5) and especially docosahexaenoic (22:6) n-3 hydroperoxy and hydroxy derivatives specifically antagonize the platelet aggregating effect to arachidonic acid (AA, 20:4n-6) but not that of ADP or collagen. Chain length (22-C > 20-C), double-bond position (n-3 > n-6), and double-bond number (6 > 5 > 4) influence the degree of inhibition of AA-induced aggregation of human platelets. Moreover, significant differences in potency were associated with specific structural aspects of 22:6n-3 lipoxygenase derivatives of 22:6n-3 as follows: functional group (OOH > OH) and positional isomer (14-OOH, 14-OH, 20-OOH > 11-OOH, 17-OOH > 10-OOH > 11-OH, 8-OOH, 7-OOH > 4-OOH). PMID- 8729141 TI - Blood pressure, serum lipids, and fatty acids in populations on a lake-fish diet or on a vegetarian diet in Tanzania. AB - Major risk factors for coronary heart disease were assessed in two populations of Tanzania, one on a fish diet (FD) living along the coast of Lake Nyasa, and the other, mainly on a vegetarian diet (VD), living in a farming area. Lower blood pressure values were found in the FD subjects (n = 618) vs. VD (n = 618) (systolic blood pressure, SBP, 120 +/- 15 vs. 135 +/- 20, P < 0.01; diastolic blood pressure, DBP, 70 +/- 9 vs. 78 +/- 11, P < 0.01, respectively). In an FD subgroup (n = 61), total cholesterol (TC) (122 vs. 136 mg/dL, P < 0.01); triglycerides (TG) (82 vs. 105 mg/dL, P < 0.01); and lipoprotein (a) [Lp(a)] (19.9 +/- 18.4 vs. 32.3 +/- 22.4, P < 0.001) were lower than in a VD subgroup (n = 55). Serum fatty acids (FA) in the FD subgroup were as follows: eicosapentaenoic acid (EPA) (20:5) 2.48 vs. 0.72%, docosahexaenoic acid (DHA) (22:6) 5.93 vs. 1.49%, vs. the VD, respectively. Arachidonic acid (AA) (20:4n-6) also was higher in the FD vs. the VD group (9.85 vs. 8.30%, P < 0.05), whereas 18:2n-6 was about double (23.97 and 14.85%) in VD vs. FD. The peculiar serum FA pattern in FD reflected the FA of dietary fish. In fact, in four main species of lake fish, DHA was 8-19%, higher than EPA (1.8-4.2%), in contrast with the situation in cold-water fish, and AA was 5.8-8%, higher than in cold-water fish. The data, obtained in populations strictly on natural, unprocessed, low-fat diets, show that a diet based on freshwater fish results in lower BP, serum TC, TG, and Lp(a), and suggests that serum AA is not reduced when the major dietary n 3 is DNA rather than EPA. PMID- 8729142 TI - n-3 fatty acids decrease colonic epithelial cell proliferation in high-risk bowel mucosa. AB - The n-3 fatty acids (C20:5, eicosapentaenoic acid; c22:6, docosahexaenoic acid) may be important in the development, growth, and metastasis of colon cancer, a leading cause of death in North America. Patients who have had a bowel neoplasm have a high risk of developing a second neoplasm, and this risk is associated with a high percentage of cells correspond to the S phase of bromodeoxyuridine (BrdUrd) labeling in mucosal epithelial cells. To determine the effect of n-3 fatty acid supplementation on DNA synthesis of rectal mucosa, patients with stage 1 or stage 2 colon carcinoma or adenomatous polyps were randomized to consume either 9 g/d n-3 fatty acid capsules or 9 g/d placebo capsules. Plasma phospholipid fatty acid analysis and proctoscopic mucosal biopsies were performed at baseline, 3, and 6 mon. Colonic crypts were isolated from the mucosa, disassociated with enzymes, and incubated with BrdUrd, and %S phase was measured by flow cytometry. The plasma phospholipid n-6/n-3 ratio was determined by gas chromatography. Supplement compliance was assessed by plasma phospholipid n-6/n-3 ratio. Mean capsule consumption in these two group was 82%. Prior to supplementation, there were no significant differences in the %S phase and the plasma n-6/n-3 ratio between these groups. Patients whose colonic epithelial cells indicated hyperproliferation at baseline showed a strongly positive correlation to the %S phase of BrdUrd uptake and the n-6/n-3 ratio. There was no significant change after n-3 treatment in patients with low baseline. Those in the placebo group showed no significant difference in n-6/n-3 ratio, although there was an increase in the %S phase of BrdUrd uptake at 6 mon. The n-3 group did not have significant side effects, and polyps were not found after completing 12 mon of n-3 fatty acid supplementation. This study suggests that n-3 fatty acid may be a useful chemopreventive agent in some patients as reflected in a plasma biomarker of colon tumor growth and metastasis. A low plasma phospholipid n-6/n-3 fatty acid ratio may serve as a nutritional marker that is associated with colonic epithelial cell hyperproliferation in the n-3-supplemented group as compared with the placebo group. Characteristics of mucosal proliferation at baseline may be a crucial factor for the effect of n-3 fatty acid supplementation. PMID- 8729143 TI - The role of internal medicine in addiction medicine. AB - Internal Medicine must play an important role in addiction medicine. Although Psychiatry has been the dominant discipline in the addictions field, this article explores how the internist can bring special insight to the understanding of addictions. The article documents how Internal Medicine, historically, has helped define and diversify the field of addiction medicine. A survey of recent history, however, shows that Internal Medicine has only minimally broadened its role in the addiction field, despite the pronouncements of numerous committees, despite the urging of several medical groups that it do so. The article shows that some internists want to play an increased role in the field of addiction medicine, but often feel limited by their lack of knowledge and by the professional biases they confront. In conclusion, new suggestions for improving internist education and increasing their interest and participation in addiction medicine are offered. PMID- 8729144 TI - Gender differences in medical presentation and detection of patients with a history of alcohol abuse or dependence. AB - Women and men with alcohol use disorders differ in many respects. A retrospective medical record review of 132 patients was performed to determine outpatient clinic utilization, presentation patterns and physician actions related to patient gender and lifetime DIS-status. Women, irrespective of DIS-status, utilized out-patient health care services more often than did DIS-positive or negative men. Of 16 specific alcohol-related complaints, gender differences were only detected for trauma. DIS-positive men were more likely to have had an alcohol history taken during the preceding 12 months than were DIS-positive women. Sedatives/minor tranquilizers were most often prescribed to DIS-positive women. This study supports the need in primary care settings for more screening to detect and diagnose alcohol-abusing patients, particularly women. Physicians should routinely screen for alcohol abuse before prescribing sedatives or minor tranquilizers. PMID- 8729145 TI - A path analysis of gender differences in adolescent onset of alcohol, tobacco and other drug use (ATOD), reported ATOD use and adverse consequences of ATOD use. AB - Gender differences in the ways a risk factor approach explains onset of using alcohol, tobacco and other drugs (ATOD), reported use of ATOD and adverse consequences from ATOD use were investigated by means of separate path analyses for male and female adolescents. A fully recursive model was specified in which alienation, trait anger, interaction anxiety and cognitive motivation for ATOD use comprised the first column variables which were used to predict earliness of onset. For the second step, column one variables plus onset were used to predict ATOD use. In the final step, column one variables plus onset and ATOD use were used to predict adverse consequences of ATOD use. An ex post facto design was used in surveying 796 high school students in a single Midwestern community. The data from this sample supported the predictive validity of the theoretical model. In addition to the expected indirect or mediated effects of the risk factors via onset of ATOD use on ATOD use and via onset and ATOD use on adverse consequences of ATOD use, there were direct effects of column on risk factors on both ATOD use and on adverse consequences of ATOD use for both males and females, lending support to the concept of multiple pathway risk factors. The three gender differences that emerged in the separate path analyses were entirely consistent with the gendered deviance model. PMID- 8729146 TI - Cohabiting relationships and sexual interest of male alcoholics. AB - This study comprises data from a consecutive series of 52 heterosexual male alcoholics. The purpose of the study was to describe sexual interest and cohabiting relationships and to analyse the influence of personality and biological factors on these phenomena. An increased sexual interest or activity and frequent changes of cohabiting partners are two common characteristics of male alcoholics but these two phenomena are not interrelated. A decreased sexual interest is reported by only a minority of male alcoholics. The alcoholics with several previous cohabiting relationships stand out as being aggressive and hard drinking. The alcoholic with an increased sexual interest makes an opposite impression. He is non-aggressive, socially adjustable and unafraid of close interpersonal relations. When decreased sexual interest or activity occurs in male alcoholics, it is mainly a result of hormonal changes. Low levels of male and high levels of female sex hormones are common in these men. PMID- 8729147 TI - Alcohol and drug abuse in an urban trauma center: predictors of screening and detection. AB - Studies of the prevalence of alcohol and drug abuse among trauma patients have produced recommendations that all trauma patients be screened for these substances. But a national survey of trauma centers showed that just 72.0% of level 1 trauma centers routinely obtain blood alcohol levels on trauma patients and only 50.4% routinely utilize toxicology screens. Utilizing patient summary data collected on 2,246 trauma patients treated in the Emergency Department of Long Beach (CA) Memorial Medical Center over a 2.5 year period, and laboratory results for a random sample of 246 of these patients, this study sought to determine patient and presenting problem characteristics associated with screening and positive test results in centers (such as ours) where screening is done selectively. Results show that 75.4% of trauma patients are screened for blood alcohol levels, and 58.5% are given a urine toxicology, with lowest rates for 12-20 year olds and highest rates for patients over 40 who were confused or incomprehensible. Men were slightly more likely to be screened for blood alcohol than women, but their positivity rate was 2.5 times higher; by contrast, the toxicology positivity rate did not differ significantly by gender. A positive finding for either test was associated with a 40% higher rate of positivity for the other. PMID- 8729148 TI - The reactions of patients and doctors in the united arab emirates towards smoking. AB - One hundred male U.A.E. nationals (50 smokers and 50 non-smokers) rated their own risk for developing heart disease, developing arthritis and having an automobile accident; and the risks facing the average smoker and average non-smoker. Smokers in the U.A.E., like smokers in the U.K., exhibited "constrained optimism." Non smokers, unlike non-smokers in the U.K., were "pessimistic" about their chances of developing heart disease and having an accident. Eighty-two male doctors, 41 smokers and 41 non-smokers, were asked to respond to two hypothetical patients (one with long-term obesity newly diagnosed as being hypertensive, the other a long-term smoker newly diagnosed as having lung cancer). Compared with smoker doctors, doctors who themselves did not smoke expected to feel more uncomfortable when managing the smoker patient, and perceived him as more responsible for his condition. Possible reasons for these results, their implications and ways of pursuing the issues raised are discussed. PMID- 8729150 TI - Selective guide to current reference sources on topics discussed in this issue. PMID- 8729149 TI - Successful adherence to observed prophylaxis and treatment of tuberculosis among drug users in a methadone program. AB - Incomplete antituberculous chemoprophylaxis and treatment are major causes of the resurgence of tuberculosis, often drug-resistant, among drug users. We offered directly observed antituberculous chemoprophylaxis (n = 102) or treatment (n = 12) to tuberculous chemoprophylaxis (n = 102) or treatment (n = 12) to eligible methadone maintenance treatment patients. Methadone dosing was not contingent upon ingestion of antituberculous medication(s). No material incentives were provided. Ninety (88%) prophylaxis and 9 (75%) treatment patients were administered > or = 5 weekly doses of antituberculous medications during > or = 80% of 4740 patient-weeks. The majority of patients were HIV-seropositive. Active substance abuse was not associated with diminished adherence. Over 80% of patients completed or were still receiving therapy at the end of the study. Adherence to and completion of directly observed antituberculous therapy can thus be attained by drug users in treatment, despite ongoing drug misuse. Substance abuse treatment programs provide opportunities for enhanced compliance, and should thus be viewed as critical components of strategies to address the tuberculosis epidemic in drug users. PMID- 8729151 TI - Excess mortality and its relation to hypertension and proteinuria in diabetic patients. The world health organization multinational study of vascular disease in diabetes. AB - OBJECTIVE: To determine the extent that mortality in IDDM and NIDDM patients is in excess of that of the general population and examine its relation to hypertension and proteinuria in diabetic patients. RESEARCH DESIGN AND METHODS: A stratified random sample of 4,714 diabetic patients aged 35-55 years participating in the World Health Organization Multinational Study of Vascular Disease in Diabetes has been followed up from 1975 to 1987. Excess mortality, compared with the background population, was assessed in terms of standardized mortality ratios (SMRs) for each of the 10 international cohorts. The relationship between excess mortality and proteinuria/hypertension was examined by diabetes type and sex. RESULTS: SMRs were in general higher in patients with IDDM (ranging from 188 to 686 for men and from 336 to 790 for women) than with NIDDM (from 138 to 370 for men and from 126 to 435 for women). For both diabetes types and in both sexes, SMRs decreased with increasing age and increased with increasing diabetes duration. Patients with both hypertension and proteinuria experienced a strikingly high mortality risk: 11-fold for men with IDDM and 18 fold for women with IDDM and 5-fold for men with NIDDM and 8-fold for women with NIDDM. Even in the absence of proteinuria and hypertension, SMRs were significantly increased in both IDDM (284 men and 360 women) and NIDDM (192 men and 236 women) patients. CONCLUSIONS: Considerable international differences were found not only in mortality rates for the two types of diabetes but also in the extent of excess mortality among centers. IDDM patients had a high excess mortality in comparison with the general population. The significant excess mortality was demonstrated even in patients without proteinuria and without hypertension for both sexes and diabetes types. PMID- 8729152 TI - Glycemic control and prognosis in type I diabetic patients with microalbuminuria. AB - OBJECTIVE: To investigate the course of microalbuminuria during the 1980s in type I diabetes patients. RESEARCH DESIGN AND METHODS: This was a 10-year follow-up of 109 patients in whom type I diabetes was diagnosed between 1961 and 1980 before 15 years of age and who were initially investigated between 1977 and 1983 after a diabetes duration of > or = 3 years. Microalbuminuria was defined as an albumin excretion rate (AER) of 20-200 micrograms/min in two of three consecutive urine samples. RESULTS: At the initial investigation, 81 patients had normal AER, 27 had microalbuminuria, and 1 had macroalbuminuria. Between 1989 and 1992, 99 (91%) patients were reinvestigated. Only 5 (19%) of the initially microalbuminuric patients developed macroproteinuria during the 10-year follow-up period, and in 15 (58%) patients, AER decreased to normal. Three (4%) of the normoalbuminuric patients developed microalbuminuria but none macroproteinuria. The initially microalbuminuric patients, in whom AER normalized, improved their glycemic control from 1980-1983 to 1989-1991 (mean +/- SE HbA1c 7.5 +/- 0.2 to 6.6 +/- 0.3%; P = 0.01). CONCLUSIONS: In the majority of patients with microalbuminuria in whom it is possible to obtain good glycemic control, microalbuminuria will disappear and the risk of developing nephropathy will be markedly reduced. PMID- 8729153 TI - Psychiatric disorder and metabolic control among youths with IDDM. A longitudinal study. AB - OBJECTIVE: To investigate the longitudinal relationship between psychiatric diagnostic variables and metabolic control among youths with IDDM. RESEARCH DESIGN AND METHODS: A group of 88 youths, 8 to 13 years old at onset of IDDM, were evaluated repeatedly during a 9-year follow-up period, on average, using a standardized psychiatric protocol. Levels of HbA1 were also assessed repeatedly. Psychiatric diagnoses were derived independently of HbA1 values. RESULTS: In univariate longitudinal analyses, the psychiatric diagnosis of noncompliance with medical treatment was significantly related to HbA1 level. There was a trend of an association between any major psychiatric disorder, as well as nondepressive disorder, and HbA1. Interaction terms between IDDM duration (or age) and psychiatric variables were also significantly related to metabolic control. According to the final multivariate model of repeatedly assessed HbA1, noncompliance with medical treatment (irrespective of IDDM duration) and the interaction between nondepressive psychiatric disorder and IDDM duration contributed to worse metabolic control. CONCLUSIONS: We found some support for the hypothesis that psychiatric morbidity negatively affects blood glucose regulation and that its consequences are more marked the longer young patients have had IDDM. We did not confirm the hypothesis that depressive illness has particularly deleterious consequences on metabolic control. Noncompliance with medical treatment and having had nondepressive psychiatric illness in interaction with IDDM duration account for a statistically significant but clinically modest amount of variability in HbA1 over time. The weak relationship among these variables may explain the inconsistent findings in the literature regarding psychiatric morbidity and metabolic control. PMID- 8729154 TI - Reduction in severe hypoglycemia with long-term continuous subcutaneous insulin infusion in type I diabetes. AB - OBJECTIVE: To compare the incidence of severe hypoglycemia in patients crossed over from multiple daily injections (MDIs) of insulin to continuous subcutaneous insulin infusion (CSII). RESEARCH DESIGN AND METHODS: From a population of 225 patients using CSII, all patients who met the following selection criteria were included in the present study: 1) a minimum of 12 months on intensive therapy with MDIs before switching to CSII, and 2) a minimum of 12 months on CSII after crossover. Glycemic control and adverse event rates for the 1-year MDI control period were compared with those for the CSII therapy period. RESULTS: The incidence of severe hypoglycemia during MDI therapy declined from 138 to 22 events per 100 patient-years during the 1st year of CSII (P < 0.0001) and remained significantly lower in years 2, 3, and 4 on CSII (26, 39, and 36, respectively). HbA1c levels did not change significantly between the MDI phase and any year on CSII. However, in the subgroup of patients who had pre-CSII HbA1c levels of > or = 8.0%, the change to CSII was associated with a significant reduction in HbA1c from baseline to year 1 (8.9 +/- 0.8 vs. 8.1 +/- 1.0%, P = 0.0004). The difference in diabetic ketoacidosis rates between the MDI year (14.6 events per 100 patient-years) and the CSII period (7.2 events per 100 patient years) was not statistically significant. CONCLUSIONS: CSII therapy was associated with a marked and sustained reduction in the rate of severe hypoglycemia without adversely affecting the level of glycemic control attained during MDI therapy. The more reproducible and flexible insulin delivery afforded by CSII was considered to be the major factor contributing to the improvement in severe hypoglycemia rates. PMID- 8729155 TI - Time-space clustering of date at birth in childhood-onset diabetes. AB - OBJECTIVE: To investigate whether there was a temporal and geographical clustering of time of birth for infants with childhood-onset diabetes. RESEARCH DESIGN AND METHODS: The nationwide Swedish Childhood Diabetes Registry, which ascertains 99% of children with recent-onset diabetes (0-14 years), was linked with the Swedish Medical Birth Registry. Clustering of 3,725 patients as to place and time of birth was studied compared with the general population. For each municipality (and in the three large cities of Sweden for each parish), the observed number of patients was compared with the expected number calculated from the average total rate and the number of births in that municipality. Clustering in time of birth within municipality was analyzed using a modification of a set technique by Chen (14). RESULTS: There was no consistent variability in diabetes risk by calendar birth month, but for specific years, the risk varied during the year. When geographic localization for place of birth was studied on a municipality level, four municipalities showed a statistically significant case excess while one would have been expected by chance. When we looked for clusters in both time and space for date of birth, clearly more clusters than expected were identified (P < 0.01). Of the total of 198 primary clusters, 42 included three or more patients being born in the same municipality within an unlikely short period always < 2 years. CONCLUSIONS: This is the first study indicating a clustering according to place and time of birth for later risk to develop type I diabetes. Such a phenomenon would agree with the hypothesis that infections in early life, including fetal infections, can increase the risk for diabetes. PMID- 8729156 TI - Factors associated with glucose and insulin levels in healthy postmenopausal women. AB - OBJECTIVE: Little is known about the covariates of hyperglycemia and hyperinsulinemia. We examined candidate factors in postmenopausal women. RESEARCH DESIGN AND METHODS: We determined the cross-sectional associations of sociodemographic, body-size, lifestyle, reproductive, and menopausal factors with pretrial fasting and postchallenge glucose and insulin levels in 869 postmenopausal women aged 45-65 years. Women were participants in the Postmenopausal Estrogen/Progestin Interventions study who were not taking estrogen or insulin. RESULTS: Plasma glucose levels increased significantly with age; serum insulin levels did not. BMI and waist-to-hip ratio (WHR) each showed graded positive and independent associations with glucose and insulin levels. Alcohol intake, cigarette smoking, physical activity, parity, education, and income were also associated with insulin or glucose in age-adjusted models. In multivariable models, BMI and WHR explained 18% of the variability in fasting glucose, 16% in postchallenge glucose, 28% in fasting insulin, and 17% in postchallenge insulin. Age and all other factors combined accounted for < 6% of the variance in glucose or insulin. In multiply adjusted models, African-American and Hispanic women had higher fasting and 2-h insulin levels than non-Hispanic white women. CONCLUSIONS: Most of the variance in glycemia and insulin is unexplained. Measures of obesity and fat distribution account for nearly all the explained variance. PMID- 8729157 TI - Effect of exercise intensity on glucose and insulin metabolism in obese individuals and obese NIDDM patients. AB - OBJECTIVE: The primary purpose of this study was to evaluate the acute effect of exercise of differing intensity on plasma glucose and insulin responses to an oral glucose challenge. RESEARCH DESIGN AND METHODS: Six obese men and six obese men with NIDDM of similar age, weight, percentage body fat, and VO2peak participated in the study. Each subject underwent two 7-day exercise programs in a counterbalanced order at 2-week intervals. During each 7-day exercise period, the subjects cycled every day at a power output corresponding to 50% VO2peak for 70 min or 70% VO2peak for 50 min. Muscle glycogen utilization was estimated during exercise on day 7 using a [3H]glucose infusion technique in conjunction with indirect calorimetry. During the day before and after each 7-day exercise period, a 3-h oral glucose tolerance test (OGTT) was administered after a 12-h overnight fast. RESULTS: The average caloric expenditure did not differ between exercise at 50 and 70% VO2peak in both obese and obese NIDDM subjects. However, the carbohydrate oxidation was higher (P < 0.05) during exercise at 70 than 50% VO2peak in obese subjects (77 +/- 5 vs. 68 +/- 6 g) and obese NIDDM subjects (70 +/- 4 vs. 58 +/- 6 g). Muscle glycogen utilization was also higher (P < 0.05) during exercise at 70 than 50% VO2peak in obese subjects (59 +/- 9 vs. 30 +/- 7 g) and in obese NIDDM subjects (48 +/- 5 vs. 24 +/- 5 g). In obese subjects, plasma glucose response area during the OGTT did not change after 7 days of exercise at either 50 or 70% VO2peak. Plasma insulin response area during the OGTT also did not change after 7 days of exercise at 50% VO2peak. However, plasma insulin response area was reduced (P < 0.05) after 7 days of exercise at 70% VO2peak (9,644 +/- 1,783 vs 7,538 +/- 1,522 microU.ml-1.180 min-1). In obese NIDDM subjects, both plasma glucose and insulin response areas during the OGTT did not decrease after 7 days of exercise at either 50 or 70% VO2peak. CONCLUSIONS: It is concluded that the exercise-induced improvement in insulin sensitivity is influenced by exercise intensity in obese individuals. The improved insulin sensitivity after 7 days of exercise at 70% VO2peak in obese individuals may be related to greater muscle glycogen utilization during exercise. The lack of improvement in glucose tolerance and insulin sensitivity after 7 days of exercise at either 50 or 70% VO2peak in obese NIDDM patients may be due to the fact that the NIDDM patients selected in the present study were relatively hypoinsulinemic. PMID- 8729158 TI - Use of dermagraft, a cultured human dermis, to treat diabetic foot ulcers. AB - OBJECTIVE: To assess the effect of a tissue-engineered human dermis (Dermagraft) in healing diabetic foot ulcers. RESEARCH DESIGN AND METHODS: This controlled prospective multicenter randomized single-blinded pilot study evaluated healing over a 12-week period in 50 patients with diabetic foot ulcers. These patients were randomized into four groups (three different dosage regimens of Dermagraft and one control group). All patients received identical care except for the use of Dermagraft tissue. Ulcer healing was assessed by percentage of wounds achieving complete or 50% closure, time to complete or 50% closure, and volume and area measurements. RESULTS: Ulcers treated with the highest dosage of Dermagraft, one piece applied weekly for 8 weeks (group A), healed significantly more often than those treated with conventional wound closure methods; 50% (6 of 12) of the Dermagraft-treated and 8% (1 of 13) of the control ulcers healed completely (P = 0.03). The percentage of wounds achieving 50% closure was also significantly higher (75 vs. 23%; P = 0.018), and the time to complete or 50% closure was faster (P = 0.056). The group A regimen was more effective than other treatment regimens. All three were better than the control, however, and a dose response was observed. There were no safety concerns. After a mean of 14 months of follow-up (range 11-22 months), there were no recurrences in the Dermagraft healed ulcers. CONCLUSIONS: Dermagraft was associated with more complete and rapid healing in diabetic foot ulcers. The recurrence data may indicate an improved quality of wound healing. PMID- 8729159 TI - Routine perioperative dipyridamole 201Tl imaging in diabetic patients undergoing vascular surgery. AB - OBJECTIVE: To assess the utility of dipyridamole thallium testing in symptomatic and asymptomatic patients with diabetes undergoing vascular surgery. RESEARCH DESIGN AND METHODS: Dipyridamole 201Tl myocardial scintigraphy was performed preoperatively in 93 consecutive patients with diabetes undergoing peripheral vascular procedures. The utility of clinical and thallium variables in predicting cardiovascular complications was assessed. RESULTS: Two groups of patients were identified: group A (36 patients) without clinical evidence of cardiac disease and group B (57 patients) with clinical evidence of cardiac disease. Dipyridamole thallium scans were abnormal in 21 of 36 (58%) of group A patients compared with 53 of 57 (93%) of group B patients (P < 0.0001). Compared with group B patients with perfusion defects, group A patients with perfusion abnormalities tended to have fewer defects per scan (2.7 +/- 1.5 vs. 3.6 +/- 1.9, P = 0.05). No perioperative cardiac complications occurred in group A patients while perioperative cardiac complications occurred in 9 of 57 (16%, 95% CI 7-28%) group B patients (P = 0.01). For the entire study population, the complication rate was 10%. CONCLUSIONS: Diabetic individuals without clinical markers for coronary artery disease appear to be at low risk for adverse postoperative cardiac events after vascular surgery. Preoperative myocardial perfusion imaging may add little to cardiovascular risk assessment in this subgroup of patients with diabetes. PMID- 8729160 TI - Neurophysiological evidence for altered higher brain functions in NIDDM. AB - OBJECTIVE: To investigate neurophysiological alterations of higher brain function in patients with NIDDM. RESEARCH DESIGN AND METHODS: Auditory P300 event-related potentials were recorded in 60 NIDDM patients who had no evidence of stroke, dementia, or any other neurological illnesses. The P300 wave latencies in diabetic patients were compared with those in neurologically healthy control subjects, with consideration of clinical parameters and diabetic complications. RESULTS: Diabetic patients had significantly longer P300 latencies than control subjects. Seven diabetic patients had pathologically prolonged P300 values. There was a trend toward longer P300 latencies in diabetic patients with retinopathy and in those with HbA1 of > or = 10%; the differences between these diabetic subgroups and control subjects were both statistically significant. However, three of seven diabetic patients with abnormal P300 latencies had no retinal lesions, and no significant correlation was observed among diabetic patients between P300 latencies and HbA1 levels. Peripheral neuropathy, nephropathy, blood glucose levels, and disease duration appeared not to correlate with P300 alterations. CONCLUSIONS: These findings suggest the presence of central nervous pathological processes in NIDDM that affect higher brain functions, as assessed by P300 latencies. Our findings also suggest that microangiopathy and metabolic derangement during the preceding 4- to 8-week period may contribute in small part to the pathophysiology of this central nervous involvement. PMID- 8729162 TI - Determinants of progression of microalbuminuria in adolescents with IDDM. AB - OBJECTIVE: To evaluate the significance of microalbuminuria in adolescents with IDDM and to study the relative importance of blood pressure (BP), metabolic control, and albumin excretion rate (AER) on progression of microalbuminuria. RESEARCH DESIGN AND METHODS: A cohort of 155/156 children and adolescents followed from onset up to 18.3 years of IDDM participated. In a previous follow up in July 1991 (up to 15 years of duration), 17 patients had developed persistent microalbuminuria (> or = 20 micrograms/min). In these adolescents, we analyzed whether microalbuminuria had progressed (in mean > or = 5% per year), had remained unchanged, or had normalized (< 20 micrograms/min) after another 3 years. The predictive values of mean HbA1c, diastolic blood pressure (dBP), systolic blood pressure (sBP), overnight AER, sex, age, and duration of diabetes for the progression of microalbuminuria were determined using multiple regression modeling. RESULTS: Seven of 17 patients with microalbuminuria in July 1991 had normalized, 6 of 17 patients had progressed, and 4 of 17 patients had remained unchanged after 3 years. Progressors had higher mean HbA1c during the first 5 years of IDDM and higher mean sBP in 1991 than nonprogressors. Patients with normalized microalbuminuria all had AER < 30 micrograms/min in 1991, were younger at onset of microalbuminuria, had lower mean HbA1c, and had lower dBP before normalized AER than nonregressors at the same duration of microalbuminuria. In multivariate analysis, independent significant predictors for progression were first 5-year mean HbA1c, mean AER, and mean sBP in 1991 (R2 = 0.76, P = 0.001). CONCLUSIONS: Progression of microalbuminuria in adolescents with IDDM is predicted by early sustained hyperglycemia, later elevated sBP, and increased AER per se. Microalbuminuria is frequently normalized in adolescents, and this is associated with better prevailing metabolic control, younger age, and lower dBP. PMID- 8729161 TI - Serum lipid and lipoprotein metabolism after glucose ingestion in NIDDM and IGT patients. Apolipoprotein-containing particles. AB - OBJECTIVE: Through analysis of apolipoprotein-containing particles after glucose ingestion, we determined abnormal lipid metabolism in NIDDM patients and subjects with impaired glucose tolerance (IGT) without obesity or hyperlipidemia. RESEARCH DESIGN AND METHODS: Apolipoprotein (apo) AI- or B100-containing particles or remnant-like particles (RLPs) were isolated by immunoaffinity columns prepared with monoclonal antibodies. The cholesterol contents in these particles after glucose ingestion (0, 30, 60, and 120 min) were analyzed in 18 nonobese NIDDM patients, 18 nonobese IGT subjects, and 33 normal control subjects. RESULTS: Changes in cholesterol content in apo B100 particles after oral glucose ingestion decreased significantly in both the NIDDM group and the IGT group; the changes were insignificant in the nondiabetic group. Changes in cholesterol content in the apo AI particles after oral glucose ingestion showed no differences between the three groups. Changes in cholesterol content in RLPs after oral glucose ingestion increased significantly in both the NIDDM group and IGT group; the changes were insignificant in the nondiabetic group. The changes in apo B100 after oral glucose ingestion decreased significantly in the three groups; however, apo AI after oral glucose ingestion showed no change in the three groups. CONCLUSIONS: The cholesterol content in apo B100 particles might decrease and the cholesterol content in RLPs might increase after oral glucose ingestion in not only NIDDM patients but also IGT subjects. The changes may be partially due to insulin resistance. PMID- 8729163 TI - Postural rearrangement in IDDM patients with peripheral neuropathy. AB - OBJECTIVE: To evaluate the influence of diabetic peripheral neuropathy on postural strategy. RESEARCH DESIGN AND METHODS: Static posturography and nerve conduction velocity were performed in the following age-matched subjects: 10 IDDM patients with peripheral neuropathy, 23 IDDM patients without peripheral neuropathy, and 21 control subjects. All subjects with signs or symptoms of postural instability were excluded from the study. The following posturographic parameters were drawn: 1) velocity of body sway, expressed as mean velocity and average of the SDs, 2) VFY, the parameter derived from the velocity variance and the anteroposterior mean position of the body (this parameter monitors the postural strategy pursued by the subject), and 3) fast Fourier transformation on the x (FFTX) and y (FFTY) planes, spectral analysis of the frequencies of body oscillation on frontal (x) and anteroposterior (y) planes. RESULTS: Mean velocity and its SD were higher in IDDM patients with peripheral neuropathy than in control subjects and IDDM patients without peripheral neuropathy (P < 0.001). VFY was increased in IDDM patients with peripheral neuropathy versus control subjects and IDDM patients without peripheral neuropathy (P < 0.01). A direct relationship was found between parameters of posturography and some parameters of nerve conduction tests. CONCLUSIONS: Diabetic patients with peripheral neuropathy demonstrate a shift from physiological ankle control to hip postural control as monitored by specific posturography analysis. PMID- 8729164 TI - A case of myocarditis associated with IDDM. AB - We report a case of diabetic ketoacidosis (DKA) complicated by acute myocarditis, which was confirmed by cardiac biopsy. A 26-year-old man was hospitalized with severe DKA. On admission, nonspecific ST-T change was noted on the electrocardiogram (ECG). The patient's levels of creatine phosphokinase (CPK) and glutamic oxaloacetic transaminase were slightly elevated, but he did not complain of chest discomfort or symptoms of heart disease. On the first day after admission, ST-T elevation was noted on ECG during treatment of DKA. By cardiac angiography and cardiac biopsy, coronary heart disease was ruled out and postmyocarditic change was histologically confirmed. An episode of upper respiratory viral infection before the onset of acute diabetes suggested that the patient suffered from viral-induced myocarditis and consequent development of IDDM. This possibility was confirmed by the clinical course of ECG change, with elevated CPK and lactate dehydrogenase and a slightly elevated antibody titer for echovirus. PMID- 8729165 TI - Milk and type I diabetes. PMID- 8729166 TI - Short-term cost benefit of pre-conception care for diabetes. PMID- 8729167 TI - Pump-pocket complications during long-term insulin delivery using an implanted programmable pump. PMID- 8729168 TI - Fatal toxic intrahepatic cholestasis secondary to glibenclamide. PMID- 8729169 TI - Insulin resistance in the pathogenesis of dyslipidemia. PMID- 8729170 TI - Acute effects of insulin in the control of VLDL production in humans. Implications for the insulin-resistant state. AB - The role of hyperinsulinemia in the pathogenesis of triglyceride (TG) and VLDL over-production in insulin-resistant states remains controversial. While studies in humans and animals have generally suggested that chronic hyperinsulinemia facilitates VLDL production, particularly in the presence of an abundant supply of substrate for VLDL synthesis, the majority of in vitro studies using cultured hepatocytes and hepatoma cell lines have demonstrated an acute inhibitory effect of insulin. Using radiolabeled VLDL tracers we have examined the acute effect of hyperinsulinemia on VLDL production in humans. We found a rapid suppression of plasma free fatty acid (FFA) levels in response to insulin and a consistent 50 60% insulin-induced suppression of both VLDL TG and VLDL apolipoprotein (apo) B in lean insulin-sensitive individuals. Elevation of plasma FFA levels by infusing heparin and Intralipid without hyperinsulinemia resulted in a marked increase in VLDL TG and VLDL apoB production. When the insulin-induced suppression of plasma FFA levels was prevented during hyperinsulinemia, VLDL TG production was still inhibited, although to a lesser extent than with insulin alone. We concluded from these findings that insulin suppresses VLDL production in insulin-sensitive humans partly by suppressing plasma FFA levels and partly by a non-FFA-mediated (perhaps direct hepatic) mechanism. In addition, we found that chronically insulin-resistant hyperinsulinemic obese individuals were resistant to this suppressive effect of insulin on VLDL apoB production, in keeping with similar findings by others performing in vitro experiments using cultured hepatocytes isolated from insulin-resistant or hyperinsulinemic rats. The relevance of these findings to the mechanism of hypertriglyceridemia associated with chronic insulin resistant states in humans remains a matter of speculation. One hypothesis is that resistance to the normal suppressive effect of insulin, in association with other metabolic abnormalities associated with insulin resistance, may contribute to postprandial and postabsorptive hypertriglyceridemia. PMID- 8729171 TI - Fatty acids and insulin resistance. AB - We have demonstrated that physiological elevations in plasma free fatty acid concentrations inhibit insulin-stimulated glucose uptake in a dose-dependent manner in normal control subjects and in patients with NIDDM. Two possible mechanisms were identified: 1) a fat-related inhibition of glucose transport or phosphorylation that appeared after 3-4 h of fat infusion and 2) a decrease in muscle glycogen synthase activity that appeared after 4-6 h of fat infusion. We conclude that elevations of plasma FFAs caused insulin resistance and hence may play a significant role in the pathogenesis of insulin resistance in obesity and NIDDM. PMID- 8729172 TI - Candidate genes for insulin resistance. AB - Insulin resistance confers increased susceptibility to NIDDM, atherosclerotic cardiovascular disease, ovarian hyperandrogenism, and possibly hypertension. Insulin resistance is largely inherited, in rare cases as a monogenic disorder or more commonly as a complex trait. The search for insulin resistance genes relies mainly on two complementary approaches: 1) positional cloning using random DNA markers present throughout the genome; and 2) the analysis of specific candidate genes. This report briefly summarizes the candidate gene approach to insulin resistance. Progress related to the analysis of genes encoding molecules that participate in insulin action is reviewed. In addition, the spectrum of potential genetic defects that might contribute to insulin resistance, both at the level of the target cell and secondarily (e.g., obesity genes), is discussed. PMID- 8729173 TI - Hypertension and nephropathy. PMID- 8729174 TI - The therapy by ischemic stress induces a durable EEG synchronization in alpha frequency band. AB - The effect of forearm or calf ischemia on computerized EEG was studied in five patients with peripheral nervous system affections and five patients with cerebral infarction. Spatial analysis of power spectra in various frequency bands was performed in two 8-sec epochs, the former before ischemia and the latter after 5-min from the cessation of circulatory arrest which lasted 15 min. Transient peripheral ischemia resulted in consistent augmentation of alpha activity involving both the anterior and the posterior hemispheric areas, bilaterally. On the other hand, the slower frequency bands (theta, delta) underwent minor, nonsignificant alterations after ischemic stress. The rise of alpha amplitude induced by ischemic stress was ascribed to the activation of nonspecific thalamic system. The activation of this system might also be involved in the favourable effects exerted by ischemic stress on motor recovery of paretic limbs. PMID- 8729175 TI - Investigation of P300 in various forms of epilepsy. AB - The problem of the late EP components generator localization has not yet been solved. This study aims to find out if a focal cortical epilepsy of the frontal (FLE) or of the temporal lobe (TLE) can modify the P300 component of the evoked response (either visual or auditory). Our data rely mainly on the selective differentiation test of an 80 dB click alternatively applied on the right or left ear, the patient having to give a motor response to those stimuli which have been chosen as target stimuli (probability of administration, 1:4). Under these circumstances, in normal subjects, evoked potentials showed deep P300 and an attenuated N2b in case of target stimulation and high, delayed N2b and an attenuated P300 for the nontarget stimulation, FLE did not abolish the late components of the EP; on the contrary, these were significantly augmented by the task. TLE, in exchange, severely attenuated differences between N2b-P300 components recorded from the scalp in case of target and nontarget stimulation. P300 was even higher at rest (possibly an effect of defective habituation). Rare or rapid stimuli application which was separately investigated gave another type of EPs modification compared to that specific for the auditory discrimination task. These facts are corroborated with the complete abolition of late EP components after lateral brain stem lesions. The conclusion is that N2a component has a reticular brain stem generator while N2b and P3 have initial generators in the temporal lobe (hippocampus). PMID- 8729176 TI - A new trend in the therapy of orthostatic arterial hypotension: prevention by propranolol or metoclopramide of the excessive adrenaline release of brainstem infarct patients with postural hypotension. AB - The catecholamine (CA) response to upright posture was studied in 30 brainstem infarct patients with orthostatic arterial hypotension; the investigation was made before and after 10 days propranolol therapy (in 15 cases) and before and after 10 days metoclopramide therapy (in other 15 cases). Before treatment almost all patients responded to posture by a rise in adrenaline (A) excretion and by a depression in noradrenaline (NA) excretion. Propranolol therapy prevented the excessive A release produced by standing and normalized their NA response to posture. Metoclopramide administration also prevented the post-orthostatic A discharge but had no significant influence on NA response to posture. Both drugs exerted a favourable influence on postural hypotension of investigated patients. As post-orthostatic A discharge observed in patients with postural hypotension is involved in the pathogeny of this syndrome and both metoclopramide and propranolol are able to correct this disorder one may maintain that the clinical favourable results obtained with these drugs are ascribable at least partly to their blocking effect on A release. PMID- 8729177 TI - The role of immune processes in amyotrophic lateral sclerosis pathogenesis. AB - Despite many efforts, the etiopathogenesis of ALS remains unknown. During the last decade evidence for an autoimmune involvement in motoneuron degeneration and death has remarkably increased. Multiple reports have documented significant expression of proteins associated with immune function in affected areas of ALS patients. Two animal models of immune-mediated motoneuron destruction have been developed that closely resemble clinical, electrophysiological and morphological features of human ALS. Inflammatory foci within the spinal cord, and IgG at the neuromuscular junction as well as within upper and lower motoneurons found in the animal models support the role of autoimmune mechanisms of motoneuron destruction in this model. IgG from ALS patients and from the animal models can passively transfer physiological changes at the neuromuscular junction in mice. That ALS IgG interact with calcium channels and induce an alteration of their function is now electrophysiologically and biochemically evident. Furthermore, it has been documented that motoneurons may be selectively vulnerable since they have a deficient calcium buffering capacity. Although further research efforts are necessary to elucidate the interaction of the ALS antibodies with the calcium channel function and how defective calcium handling by the motoneurons is important in their degeneration, the current data strongly suggest the involvement of autoimmune mechanisms in ALS etiopathogenesis. PMID- 8729178 TI - Tomaculous neuropathy with unusual clinical aspects. AB - Tomaculous neuropathy represents the morphological substrate of the recurrent familial neuropathy with liability to pressure palsies. Some ultrastructural changes characterizing the tomaculous neuropathy can occur as incidental aspects in other different neuropathies. Few tomaculous neuropathy cases with clinical aspect of chronic polyneuropathy without paretic episodes have been mentioned in the literature. In the present work, we report four cases who offered the morphological surprise of a true tomaculous neuropathy with 15-37% of the teased fibres bearing tomaculae sized: 55-106 microns/20-23 microns, on the background of a demyelinating neuropathy with 25-56% of the teased fibres showing segmental de- or remyelination. The clinical and electrophysiological diagnoses of these 4 patients were: HSMN type I (2 cases), HSMN type VIII (polyneuropathy associated with a cerebello-extrapyramidal syndrome -1 case), and a neurogenic scapuloperoneal syndrome (1 case). The specificity of the tomaculous neuropathy is discussed. PMID- 8729179 TI - Associated pathology with Wallenberg's syndrome. A report of three cases. AB - Three cases with Wallenberg's syndrome were reported, in which CT-scan revealed silent old right temporal infarction, right recent cerebellar haematoma and left calcified parietal subdural haematoma evidenced after 10 years since trauma due to a traffic accident. Although Wallenberg's syndrome constitutes a fully clinical diagnosis, CT-scan does not seem to be justified by the management logical deductions of the medical act; however, it is better to perform it in order to identify a possible associated pathology. PMID- 8729180 TI - Therapy study on the experimental injury of spinal cord. IV. High doses of methyl prednisolone. AB - The effect of large glucocorticosteroid hormones administered to two groups of adult rabbits with an experimental acute injury of the spinal cord was studied. In the former (10 rabbits), each animal received intravenous 30 mg/kg body weight of prednisolone-21-sodium-hemisuccinate after 30 minutes and further 15 mg/kg b.w. at intervals of 2, 24 and 48 hours since injury. In the latter (40 rabbits), each animal received 15 mg/kg b.w. methyl-prednisolone-21-hydrogen-sodium hemisuccinate at 30 min, 2, 24 and 48 h postinjury. A group of 20 injured rabbits with the same experimental conditions was used as controls without glucocorticoid treatment. Clinically, both treated groups of rabbits showed a significant improvement of posterior limb movements and in the general state of health compared with the controls. The mean motor score (according to a 5-score scale) indicated a significant difference between the two treated groups (2.3 and 2.6, respectively and 1.7 for the controls). Histologically, the modifications correlated with the clinical posttraumatic symptomatology and severity in all the three groups: histopathological modifications consisted in oedema, ischaemic cells, diffuse microglial hyperplasia, microhaemorrhages, all of them located in the central gray matter. In all cases with a severe clinical evolution (score of 1-3), aggravated histopathological modifications both in the gray and white matter are evidenced: necrosis; vacuolization; cavitation; myelinic and axonal fragmentation, demyelination, mesenchymo-glial scar reaction. In all groups, regenerated nervous fibres were noticed in the dense scar of the injured cord. PMID- 8729181 TI - How I do it. Arteriovenous malformation management. AB - Arteriovenous malformations (AVM) are rare vascular lesions that can present with a myriad of clinical presentations. In our institutions, initial workup consists of a clinical exam, color Doppler imaging, and magnetic resonance imaging. After the initial noninvasive workup, arteriography, at times closed system venography, and ethanol endovascular repair of the AVM is performed under general anesthesia. Depending on the size of the lesion, additional Swan-Ganz line and arterial line monitoring are performed. Patients are usually observed overnight and uneventfully discharged the following day if no complication occurs. Patients are followed at periodic intervals despite cure of their lesion. Long-term follow-up is essential in AVM management. PMID- 8729183 TI - [Manual intrauterine aspiration using a Karmann syringe. Multicenter study in Sonora and Sinaloa, Mexico]. AB - These are the results of a prospective, multicentric study carried out in five institutional hospitals in Sonora and Sinaloa, using Karmann syringe and cannulae for manual intrauterine aspiration in cases of incomplete abortion, dead and retained ovum, hidatiform mola and endometrial biopsy. Management data in 1046 patients, are described; methodology is shown, and demonstrating that it converts uterine emptying is a simple, efficacious and economical procedure, with a very low risk and it allows these patients management in an ambulatory way. From these results, and as it is demonstrated in the study, institutions may increase their programable resources as to amount of disposable beds, for other type of patients, and the offering of attention by surgeon, anesthetist, nurse and medication, as their hospital stay is minimal, and in most of the cases active anesthetist participation is not required. In the same way, it permits the patient a better relationship with the personal of the institution, and so, a more human care and the possibility of reintegrating the patient to a familiar environment, rapidly, and in good psychical and physical conditions. PMID- 8729182 TI - Retained contrast after embolization of a right gastric artery pseudoaneurysm. AB - Pseudoaneurysms due to chronic pancreatitis can be a source of major gastrointestinal (GI) hemorrhage. Computed tomography (CT) is the primary diagnostic imaging modality for pancreatic pseudocysts associated with GI bleeding. Pseudoaneurysms and associated GI bleeding can be diagnosed and embolized with transcatheter techniques once the arterial anatomy is defined. CT is a useful modality for follow-up examination of the pseudocyst; the findings must be correlated with other procedures performed on these patients. On follow up studies, contrast medium retained in the pseudocyst after embolization may falsely signal persistent bleeding into the pseudocyst. PMID- 8729184 TI - [Puerperium with short-term hospitalization]. AB - Seventy seven cases with discharge before 12 hours post-partum and 240 cases with discharge after 24 or more hours after delivery, were evaluated at Hospital General de Mexico, in an observational, prospective, cohort, study in low risk deliveries. Both groups were reviewed ten days after delivery. There were no different ratios for abnormal uterine hemorrhage, urinary tract infection or endometritis. Thrombophlebitis was more frequent in early discharge patients, both statistically and clinically significant (RR3.58, CI95% 1.48-8.67, P = 0.003). This finding is discussed, as well as measures to prevent it. It is concluded that early discharge programs in low risk conditions, are convenient. PMID- 8729185 TI - [Ethical and legal considerations on assisted human reproduction in Mexico]. AB - Assisted Human Reproduction (AHR) has been one of the most important advances in Ginecology and Obstetrics technology, it has contributed to improve understanding on deeper biological mechanisms of human reproduction. Since Louise Brown's birth, in 1978, in vitro fertilization and Human Embryos Transference (HET) developed by Steptoe and Edwards, became the basis for new AHR methods. Later, writen information has increased, so, it is easy to find a lot of scientific reports in national and international specialized journals. These reports contents not only technological advances, but a lot of ethical thoughts which had showed the necessity for a worldwide legislation. PMID- 8729186 TI - [Prevalence of rubella virus antibodies in pregnant women in two zones of Valle de Mexico]. AB - The aim of this study was to determine the prevalence of specific antibodies against rubella virus in pregnant women from Iztapalapa and Nezahualcoyotl areas of the valley of Mexico, in order to evaluate susceptibility to this virus in these areas. Serum samples collected from 5535 women between 15 and 44 years old were studied by the hemaglutination inhibition test. 92.6% of positivity was obtained and no significant difference was observed between different age groups. The prevalence of rubella antibodies in pregnant women was 92.4% for Iztapalapa zone and 92.8% for Nezahualcoyotl. The titles of antibodies against rubella virus more frecuently presented were 1:16 (36.3%) and 1:32 (29.6%). The data obtained in this study suggest that women attending Medical Services of the Mexican Institute of Social Segurity in reproductive age from the Iztapalapa and Nezahualcoyotl areas, do not need massive vaccination against rubella virus. PMID- 8729187 TI - [Value of the Q-tip test in patients with urinary incontinence]. AB - We evaluated the accuracy of the Q-tip test to measure uretrhovesical junction mobility in patient's histories of urinary incontinence, and pelvic relaxation. A retrospective study of 183 files of patients with urinary incontinence and anterior vaginal wall relaxation in the Instituto Nacional de Perinatologia (INPer) between march 1994 and may 1995. All patients underwent a systematic diagnosis evaluation with detailed history and physical examination including Q test, neurology examination, urinary, urine culture, uretrhoscopy and urodinamic studies. We correlated Q-tip test pelvic relaxation to measure urethrovesical junction mobility. They were placed in three groups. Group I formed by stres incontinence, group II formed by mixed incontinence and group III formed by recurrent incontinence. Mean age 46.3 years (29-72), almost all the patients were multiparas, menopausal status in 72, with hormonal treatment 48. Q-tip test was not different between the groups. Group I had cystocele II with hipermobility statistically significant difference, the same in group II. No differences were found in the group III. We concluded that urethral position and mobility as measure by the Q-tip test are related to defects in interior vaginal support, but not to specific urologic diagnosis. PMID- 8729188 TI - [Effect of prepuberal gonadectomy in partial androgen resistance syndrome]. AB - The androgen resistance syndrome, is a dominant and recessive linked to X disorder, with clinically heterogenous manifestations. Reported case. A 17-years patient with primary amenorrhoea and genital ambiguity. Bilateral inguinal herniplastia and probable gonadectomy were performed at 14-months-old. His cariotipe was male 46,XY and the hormonal determinations showed the presence of hypergonadotropic hypogonadismo. The congenital adrenal hyperplasia (deficiency of 21-hidroxilase) was discarded. In the partial androgen resistance syndrome, the prepuberal gonadectomy avoid a progresive virilization of the external genitals. However, the estrogenic hormonal replacement is mandatory. PMID- 8729189 TI - [Current state of conservative management of ectopic pregnancy]. AB - The ectopic pregnancy constitutes a disease that require the most diagnostic accuracy, not only for the biologic importance of its clinical manifestations, but for the sequelae of its treatment too, in relation with the fertility of a patient frequently affected with infertility so far. The increase in its frequency, by multiple factors, including treatment of infertility itself and Assisted Reproduction techniques, make the clinician must be alert and have availability of all diagnostic tools for the establishment of an oportune and conservative treatment, directed to prevent an acute alteration of the hemostasis and a shadowy reproductive future. The present review of the literature about the conservative management of the ectopic pregnancy let us have to know the criteria neccessary for giving the optimum treatment to our patients and offering them a better reproductive future. PMID- 8729190 TI - [Immunologic aspects on the etiology of pregnancy toxemia]. AB - Since 70 years ago, the possibility that the etiology of Gravidic Toxemia is inmunologic has been studied. Several researchers agree in the disminution of the levels of IgG with no modifications on the levels of complement fractions. The determination of IgA, IgM, IgC, C3 and C4 was made on fifteen pacients with gravidic toxemia and fifteen pacients that presented no complicated pregnancy. The results are the same as the reported facts like disminution of IgG, it was observed also, a disminution on the complement fractions C3 and C4. The research let us stabblish the posibility of a sikness caused by immune complexs having as an antigenic stimulations the trofoblast. PMID- 8729191 TI - [Receptor (ER, PgR) levels as prognostic factors in the endocrine therapy of pre- and post-menopausal patients with stage III infiltrative ductal and lobular cancer of the breast]. AB - Receptors group (ER+, PgR+) in post-menopausal and pre-menopausal patients with ductal and lobular infiltrating mammary cancer clinical stage III and poorly differentiated histological assessment was studied. Levels of (ER+, PgR+) were correlated to endocrine therapy response (Tamoxifen) after 5 years of its administration. Both pre and post-menopausal patients with infiltrating ductal carcinoma, the (RE+, RPg+) tumour content, the mean values were 52 +/- 8, 53 +/- 11 fmol/mg protein and 111 +/- 20, 36 +/- 7 fmol/mg protein, respectively. With regard to lobular carcinoma, the (RE+, RPg+) tumour content, the mean value was 109 +/- 28, 46 +/- 12 fmol/mg protein in pre-menopausal patients, whereas it was 287 +/- 60, 66 +/- 18 fmol/mg protein in post-menopausal patients. The tumour was considered (ER+, PgR+) when specific binding was than 10 fmol/mg protein. In both ductal and lobular carcinoma the mean (ER+) concentrations are significantly different between the post and pre-menopausal patients, while the mean (PgR+) concentrations are significantly different in post-menopausal women. After five years of treatment with tamoxifen, survival analysis of patients with ductal infiltrating and lobular infiltrating carcinoma revealed a very strong correlation between levels of receptor group (ER+, PgR+) and their response to endocrine therapy. PMID- 8729192 TI - [Variations in the molecular forms of LH and FSH during the normal ovarian cycle]. AB - FSH and LH concentrations during the menstrual cycle are well known, showing characteristically a midcycle surge in LH as well as in less degree in FSH. At the present there is an increasing interest in studying variations of the moleculoar forms of both gonadotropins. We have studied the chromatographic profile of FSH and LH in sera obtained from women regularly ovulating and in patients with anovulatory cycles by the use of gel column chromatography. It was observed the predominance of LH 32-34 kDa at midcycle and the heterogeneous FSH profile during the ovarian cycle. In sera from anovulatory patients the chromatographic profile was even more irregular for both LH and FSH. It is concluded that LH 32-34 kDa may be the more biological isoform in turn related with the process of ovulation. PMID- 8729193 TI - Purification of the glucocorticoid receptor-mineralocorticoid receptor modulator 2 from rabbit liver. AB - Modulators-1 and -2 are endogenous low-mol-wt regulators of glucocorticoid and mineralocorticoid receptors and protein kinase C. Structural analysis of apparently purified modulators suggested that these molecules were novel ether aminophosphoglycerides. Subsequent X-ray crystallography and NMR spectroscopy indicated that the ultra-large scale modulator preparations were contaminated with glutamate and aspartate, although these amino acids lacked modulator activity. In this article, we describe the purification of modulator-2 from rabbit liver cytosol and the separation of this phosphoglyceride from these amino acids. This purification was similar to the ultra-large scale version (Bodine, P.V. and Litwack, G. [1990] J. Biol. Chem. 265, 9544-9554), but involved the chromatography of trypsinized rabbit liver cytosol on the 7-L bed volume Sephadex G-15 gel-filtration column. As before, two peaks of modulator activity (modulator 1 and -2), as well as a DNA-binding inhibitor (peak-3), eluted from the gel filtration column. The resulting modulator-2 pool was incubated with glutamate decarboxylase and treated batch-wise with Dowex-50W cation-exchange resin and Chelex-100 resin. This enzyme/resin-treated modulator-2 preparation was then chromatographed on a Dowex-1 anion-exchange column. Finally, modulator-2 was purified by preparative silica TLC. This last purification step resulted in the separation of modulator-2 from glutamate, aspartate, and gamma-aminobutyrate. In summary, rabbit liver cytosol appears to be a reasonable source of modulator-2. In addition, treatment of the preparation with glutamate decarboxylase seems to facilitate the subsequent separation of modulator-2 from the contaminating amino acids. PMID- 8729194 TI - Identification of alpha 1-adrenergic receptor subtypes in human corpus cavernosum tissue and in cultured trabecular smooth muscle cells. AB - Recent pharmacological and functional studies have suggested the presence of more than one alpha-1 adrenergic receptor subtype in human corpus cavernosum (HCC). In this study, we sought to identify the alpha-1 adrenergic receptor (alpha 1-AR) subtypes expressed in HCC whole tissue and in trabecular smooth muscle subcultured from this tissue. We have utilized RNase protection assays and in situ hybridization (ISH) techniques to identify and localize these receptor subtypes. RNase protection assays of mRNA isolated from whole tissue demonstrated the presence of mRNA transcripts for three alpha 1-AR receptor subtypes (alpha 1d, alpha 1b, and alpha 1a). alpha 1d-AR and alpha 1a-AR appear to be more abundant than alpha 1b-AR. The identification and localization of mRNA for alpha 1-AR subtypes in whole tissue was demonstrated by RNA protection assays and ISH analysis. Immunocytochemical analysis of alpha 1-AR by an antipeptide antibody developed against a specific amino acid sequence derived from alpha 1d-AR subtype demonstrated specific staining of the smooth muscle cells, suggesting the expression of alpha 1d-AR subtype. In cultured HCC smooth muscle cells (HCC SMC), phenylephrine,alpha 1-AR agonist stimulated Na+/K+ ATPase activity, suggesting the presence of functional alpha 1-AR. RNase protection assay of mRNA isolated from HCC SMC grown in culture further demonstrated the presence of mRNA transcripts for alpha 1d-AR and alpha 1a-AR subtypes. ISH analysis and confocal microscopy also indicate that the SMC express the alpha 1d-AR and alpha 1a-AR subtypes. The data presented suggests that HCC and SMC derived from this tissue express at least three alpha 1-AR subtypes. Identification of these receptor subtypes should allow characterization of the functional role of these receptor subtypes in regulation of trabecular smooth muscle tone and penile detumescence. PMID- 8729195 TI - Expression of functional muscarinic acetylcholine receptor subtypes in human corpus cavernosum and in cultured smooth muscle cells. AB - Relaxation of the trabecular smooth muscle, which is necessary for penile erection, is controlled locally by neurotransmitters and vasoactive agents. The goal of this study was to identify and characterize muscarinic acetylcholine receptor (mAChR) subtypes expressed in cultured human corpus cavernosum smooth muscle cells (HCC SMC). Binding analysis with L-[benzilic-4,4' 3H(N)]quinuclidinyl benzilate ([3H]QNB) demonstrated the expression of specific muscarinic receptor binding sites in HCC SMC. Analysis of total RNA isolated from whole corpus cavernosum tissue and smooth muscle cells, by RNase protection assays, demonstrated the expression of mRNA transcripts for m1, m2, m3, and m4 mAChR subtypes in whole tissue and m2 and m4 subtypes in cultured cells. In situ hybridization with specific m2 and m4 probes further confirmed the expression of m2 and m4 mRNA transcripts in cultured cells. Carbachol (CCh), a nonselective cholinergic agonist, inhibited cAMP synthesis at low concentrations (0.1-1 microM) and stimulated cAMP synthesis at high concentrations (100 microM), in cultured HCC SMC. CCh (100 microM) further augmented forskolin (FSK), isoproterenol (ISO), and prostaglandin E1 (PGE1)-induced cAMP synthesis. These observations suggest that, in vivo, in HCC, ACh may activate m3 mAChR subtypes on endothelial cells or m2 and m4 subtypes on the SMC. Although m2 and m4 are thought to inhibit adenylate cyclase (AC), the augmentation of cAMP synthesis by high concentrations of CCh in SMC suggests an alternative mechanism of coupling to G-proteins that stimulates AC activity. These studies show that HCC tissue expresses different subtypes of mAChR (m1, m2, m3, and m4), whereas cultured HCC SMC express m2 and m4 subtypes. It is suggested that m2 and m4 receptor subtypes may play an important role in maintaining trabecular smooth muscle tone in vivo. The augmentation of FSK-, ISO, and PGE1-induced cAMP synthesis by CCh suggests possible development of a multidrug therapeutic approach to treatment of erectile dysfunction. PMID- 8729196 TI - Effects of ET antagonists (PD143296 and PD145065) on contractions in guinea pig hilar bronchus induced by endothelin-1 and its related peptide. AB - ETs-induced contractions were resistant to ET(A)-selective antagonists and believed to be mediated by activation of ETB receptors in guinea pig bronchus. In the present study, the effects of the ET antagonists, PD143296 (Ac-D-Phe-L-Leu-L Phe-L-Ile-L-Ile-L-Trp.2Na) and PD145065 (Ac-[(R)-2-10, 11-dihydro-5H-dibenzo[a, d]cyclohepten-5-yl]Gly)-L-Leu-L-Asp-L-Ile-L-Ile- L-Trp.2Na), on contractions induced by ET-1, ET-3, sarafotoxin S6c (STXc), and IRL1620 in the isolated hilar bronchus of the guinea pig were investigated. An ETA/B nonselective antagonist, PD145065 antagonized contractions induced by ET-1, ET-3, STXc, and IRL1620. Its antagonistic activity against ET-1, with pKB of 5.77 +/- 0.02 (n = 16, 3-10 microM), was significantly lower than that against ET-3, with pKB of 6.18 +/- 0.02 (n = 12, 3-10 microM), STXc, with pKB of 5.97 +/- 0.01 (n = 14, 3-10 microM), and IRL1620, with pKB of 6.80 +/- 0.04 (n = 14, 0.3-1 microM). Conversely, although a putative ETB-selective antagonist, PD143296 (10 microM) slightly but significantly antagonized the concentration-response curve of IRL1620 (pKB = 5.28 +/- 0.14, n = 6), it had no effect on ET-1-,ET-3-, or STXc induced contractions. These results suggest that ETs possibly activate ETB2 or an atypical ETB receptor subtype in guinea pig hilar bronchus. PMID- 8729197 TI - Babesiosis. PMID- 8729198 TI - Cytomegalovirus-induced alveolar hemorrhage in patients with AIDS: a new clinical entity? AB - We report five cases of alveolar hemorrhage associated with intravascular hemolysis in patients with AIDS. Cytomegalovirus was the only pathogen recovered from the lungs of these patients. There was evidence of multivisceral spread of the virus in all patients, and all had viremia. All had clinical, biological, and pathological features of pulmonary vasculitis, and the conditions of four improved with specific anti-cytomegalovirus therapy. PMID- 8729199 TI - Disseminated infection due to Actinomyces meyeri: case report and review. AB - Actinomyces meyeri is rarely isolated in cases of actinomycosis. We present a case of disseminated actinomycosis due to A. meyeri; the patient had an abscess of the lung, osteomyelitis of the tibia, and multiple skin abscesses. Cure was achieved with surgical debridement and administration of intravenous penicillin, followed by oral penicillin, for 1 year. A concomitant gram-negative bacillus, Actinobacillus actinomycetemcomitans, was also isolated. Review of the literature revealed only 26 well-documented cases of infection with A. meyeri. Male adults are mainly affected, and alcoholism is frequently the underlying condition in these patients. Associated bacteria were isolated in two-thirds of these cases. In contrast to other species of Actinomyces, A. meyeri often causes pulmonary infection and shows a tendency for hematogenous dissemination. Even though multiple organs are involved, the outcome for these patients is excellent when penicillin is administered for several months and surgical procedures are performed when necessary. PMID- 8729200 TI - AIDS-associated cytomegalovirus infection mimicking central nervous system tumors: a diagnostic challenge. AB - We reviewed cases of cytomegalovirus (CMV) infection of the central nervous system (CNS) that initially masqueraded as tumors in 37 of 543 consecutive patients infected with human immunodeficiency virus (HIV) and CMV who were seen at the Pasteur Institute Hospital and Saint-Louis Hospital (Paris) between 1992 and 1994. We detail the clinical features of three patients who presented with ring-enhanced space-occupying lesions mimicking CNS tumors. They were all profoundly immunodepressed (mean CD4 cell count, 13/mm3). Magnetic resonance imaging (MRI) showed enlargement of the spinal cord in one case, consistent with a space-occupying lesion and showing gadolinium enhancement; in the other two cases, ring-enhanced mass lesions were seen in the cerebral hemispheres. In all three cases marked edema and a mass effect were present. Image-guided stereotactic biopsies confirmed the diagnosis of CMV infection. The three patients' conditions improved with specific therapy. MRI showed enhanced focal intraparenchymal lesions consistent with marked focal necrosis, probably related to the severity of immunodepression, as HIV infection had been diagnosed several years previously. CMV infection should be considered as a cause of ring-enhanced space-occupying mass lesions in patients with HIV-1 infection. Earlier identification of these unusual tumorlike forms of CMV infection by means of MRI should result in improved outcome. PMID- 8729201 TI - Moraxella catarrhalis: pathogenic significance in respiratory tract infections treated by community practitioners. AB - We prospectively studied the pathogenic significance of Moraxella (Branhamella) catarrhalis isolated from 212 patients of community practitioners in Australia. This organism was most commonly isolated during winter and early spring, and 92% of isolates were beta-lactamase producers. On the basis of predetermined clinical and microbiological criteria, 42% of the isolates were definitely pathogenic, 7% were probably pathogenic, 21% were of indeterminate pathogenicity, and 30% were nonpathogenic. Factors associated with pathogenic significance included pneumonia or bronchitis (87% of patients), predisposing respiratory or systemic conditions (62%), isolation from sputum, and pure isolation. Thirty-six percent of patients were < 5 years old, but only 9% of isolates from these patients were pathogenic or probably pathogenic, a finding that reflects the fact that nasal-swab and nasopharyngeal-aspirate sampling is a common practice. Isolates from older patients were more likely to be pathogenically significant. An assessment of the pathogenic significance of M. catarrhalis isolated from a patient in a community practice should take into consideration factors such as the patient's age, clinical illness, and underlying conditions; the presence of other organisms; and the source of the isolate. PMID- 8729202 TI - Skull osteomyelitis due to Salmonella species: two case reports and review. AB - Osteomyelitis of the skull is a rare disease. We describe two cases due to Salmonella typhimurium and review 10 previously reported cases of salmonella osteomyelitis of the skull. This infection is frequently complicated by extradural abscess, which may be asymptomatic. Diagnostic imaging by means of computed tomographic scanning with contrast or gadolinium-enhanced magnetic resonance imaging should be performed to detect this complication. A good outcome can be expected with a combination of surgery and antibiotic therapy. PMID- 8729203 TI - Infectious etiologies of rhabdomyolysis: three case reports and review. AB - Rhabdomyolysis can be precipitated by trauma, ischemia, metabolic defects, electrolyte abnormalities, drugs, and a wide variety of infectious diseases. At our institution, recent cases of rhabdomyolysis induced by influenza prompted us to review the infectious etiologies of this entity. In addition, a thorough literature search revealed numerous case reports but no general review on this subject. This study describes representative recent cases from our institution and details the wide variety of infections that can cause muscle damage. The pathophysiological mechanisms, muscle histology, and correlation with renal dysfunction are also discussed. PMID- 8729205 TI - Randomized controlled trial of clemastine fumarate for treatment of experimental rhinovirus colds. AB - We used a rhinovirus challenge model to test the therapeutic efficacy of clemastine fumarate for reducing sneezing and nasal secretion in a double-blind, randomized clinical trial. Clemastine fumarate (1.34 mg) or placebo was administered at 8 A.M. and 8 P.M. for 4 days, beginning 24 hours, 36 hours, 48 hours, and 60 hours after viral challenge. Infected evaluable subjects who received clemastine (n = 75) had reduced sneeze-severity scores compared with those who received placebo (n = 75) on illness days 2 (0.3 vs. 0.5; P = .003), 3 (0.4 vs. 0.8; P = .0003), 4 (0.3 vs. 0.5; P = .025), and 5 (0.1 vs. 0.3; P = .03); sneeze counts for infected evaluable subjects (vs. counts for those who received placebo) were reduced on illness days 2 (1.5 vs. 3.1; P = .01), 3 (1.7 vs. 5.6; P = .0001), and 5 (0.7 vs. 1.9; P = .03). Infected evaluable subjects who received treatment had reduced rhinorrhea scores (compared with those who received placebo) on illness days 2 (0.7 vs. 1.0; P = .04) and 3 (0.6 vs. 0.9; P = .04) and had reduced nasal secretion weights on day 3 (3.6 g vs. 6.3 g; P = .03). Over 4 days of treatment, mean sneeze scores for infected evaluable subjects (vs. scores for those who received placebo) were reduced by 50%, mean sneeze counts by 57%, mean rhinorrhea scores by 27%, and mean nasal secretion weights by 35%. Other cold symptoms were unaffected by treatment. Treatment with clemastine was associated with an excess incidence of dry mouth (6%), dry nose (19%), and dry throat (17%). PMID- 8729204 TI - Increased levels of nitrite in the sera of children infected with human immunodeficiency virus type 1. AB - Nitric oxide (NO) is a newly discovered gas that plays an important role in cell communication and host resistance to infection. The production of NO was examined in the sera of seven children infected with human immunodeficiency virus type 1 (HIV-1) and in the sera of 14 children who became seronegative for HIV-1 during the first year of life. In addition, we determined serum levels of various cytokines, such as interleukin (IL)-1 beta, tumor necrosis factor (TNF)-alpha, and gamma interferon (IFN-gamma), inasmuch as these cytokines are potent inducers of NO production. Production of NO, detected as circulating serum levels of nitrite, was measured with use of the Griess reagent. Serum levels of cytokines were determined by enzyme immunoassay. Increased serum levels of nitrite were observed in children with HIV-1 infection (0.4 +/- 0.2 mumol/L; P = .013), and in those who became seronegative for HIV-1 during the first year of life (0.5 +/- 0.3 mumol/L; P = .04). Furthermore, serum levels of IL-1 beta and TNF-alpha were significantly elevated in children with HIV-1 infection (37.5 +/- 23.6 pg/mL and 91.2 +/- 45.1 pg/mL, respectively). Prophylactic administration of intravenous immune globulin provoked a significant decrease of circulating levels of nitrite in children with HIV-1 infection. In conclusion, NO may play a role as a cytostatic or cytotoxic factor for invading microorganisms, and thus it is probably involved in limiting and/or eradicating infection. PMID- 8729206 TI - Differences in outcomes for patients with bacteremia due to vancomycin-resistant Enterococcus faecium or vancomycin-susceptible E. faecium. AB - To determine the differences in outcome in cases of enterococcal bacteremia due to vancomycin-resistant organisms, we compared consecutive patients on a liver transplant service who had clinically significant bacteremia due to vancomycin resistant Enterococcus faecium (VREF) (n = 54) with a contemporaneous cohort of patients who had vancomycin-susceptible E. faecium (VSEF) bacteremia (n = 48). VREF bacteremia occurred significantly later in the hospitalization than did VSEF bacteremia (43 days vs. 24 days, respectively; P < .01); in addition, VREF was more frequently the sole blood pathogen isolated (91% of patients) than was VSEF (56% of patients) (P = .0002). Invasive interventions for intraabdominal and intrathoracic infection were required more often in the VREF cohort than in the VSEF cohort (34 of 45 patients vs. 20 of 41 patients, respectively; P = .01). Vancomycin resistance more frequently resulted in recurrent bacteremia (22 of 54 patients infected with VREF vs. 7 of 48 patients infected with VSEF; P = .006), persistent isolation of Enterococcus species at the primary site (27 of 33 patients infected with VREF vs. 7 of 18 patients infected with VSEF; P = .005), and endovascular infection (4 patients infected with VREF vs. none infected with VSEF). The decrement in patient survival, as measured from the last bacteremic episode, was greater in the VREF cohort (P = .02). Vancomycin resistance, shock, and liver failure were independent risk factors for Enterococcus-associated mortality. Higher rates of refractory infection, serious morbidity, and attributable death occurred in the VREF cohort and were partially mediated by the lack of effective antimicrobial therapy. PMID- 8729208 TI - Tuberculosis in patients infected with human immunodeficiency virus: perspective on the past decade. AB - Tuberculosis (TB) is the most common opportunistic infection and the leading cause of death in persons infected with human immunodeficiency virus (HIV) worldwide. Because HIV is spreading in regions with the highest rates of Mycobacterium tuberculosis infection, HIV is responsible for an increasing proportion of the world's cases of TB. However, advances in molecular biology, clinical practice, and public health policy during the past 5 years offer reasons for hope. Molecular methods have provided insights into the epidemiology of M. tuberculosis transmission and the mechanisms of drug resistance. Rapid diagnostic tests have been developed to facilitate the diagnosis of TB. Retrospective and prospective studies have shown that TB in the HIV-infected person is highly treatable and often preventable. Moreover, directly observed therapy can decrease rates of treatment failure, relapse, drug resistance, and secondary spread. For two consecutive years, the incidence of TB in the United States has declined. Additional resources are needed, however, to achieve similar gains in the developing world. PMID- 8729207 TI - Cavitary pulmonary lesions in patients infected with human immunodeficiency virus. AB - The differential diagnosis of cavitary pulmonary lesions in individuals infected with human immunodeficiency virus (HIV) is broad, especially in patients with advanced disease. In patients with Pneumocystis carinii pneumonia, cavitation is an uncommon manifestation of a common disease. It is unusual in patients with pulmonary cryptococcosis, coccidioidomycosis, and histoplasmosis but occurs frequently in patients with invasive pulmonary aspergillosis. In patients with pulmonary tuberculosis, cavities are more common during earlier stages of HIV disease, when cellular immunity is relatively preserved. Mycobacterium avium complex is an uncommon cause of lung disease and infrequently produces cavities. However, Mycobacterium kansasii, is often associated with cavitation. Cavities can complicate any bacterial pneumonia and are especially common with pneumonia due to Pseudomonas aeruginosa, Nocardia asteroides, and Rhodococcus equi. Noninfectious causes of cavitary lesions are rare, but cavitary lesions caused by pulmonary Kaposi's sarcoma and non-Hodgkin's lymphoma have been reported. Because of the broad differential diagnosis and because most cavities are caused by treatable opportunistic infections, a definitive diagnosis is essential. PMID- 8729209 TI - Rifabutin versus placebo in combination with three drugs in the treatment of nontuberculous mycobacterial infection in patients with AIDS. AB - The aim of this double-blind, randomized, placebo-controlled, 12-week study was to assess the efficacy of rifabutin (450 or 600 mg/d) in the treatment of disseminated nontuberculous mycobacterial infection in patients with AIDS. Companion drugs in both arms of the study were ethambutol, clofazimine, and isoniazid. Because of low accrual, the study was prematurely terminated when a total of 382 patients had been enrolled, of which 200 were eligible (i.e., their specimens were culture-positive for Mycobacterium avium complex [MAC] or Mycobacterium xenopi at baseline) and 102 were evaluable (i.e., they were eligible, were treated for a minimum of 6 weeks, and had at least one culture assessment after baseline). The original protocol called for a total of 220 evaluable patients. At week 12, rifabutin treatment was associated with higher, although nonsignificant, rates of bacteriologic conversion than was the placebo arm, with regard to both the eligible patients (25% vs. 18%) and the evaluable patients (45% vs. 38%). Corresponding median times to culture conversion were 42 vs. 63 days (eligible patients) and 43 vs. 69 days (evaluable patients). No significant difference was observed in clinical improvement, mortality, or toxicity between the two treatment arms. The addition of rifabutin to a triple drug regimen may contribute to the clearance of disseminated MAC infection in patients with AIDS, without causing additional toxicity. PMID- 8729210 TI - Unusual presentation of cryptococcosis in a patient with AIDS. PMID- 8729211 TI - Vibrio fluvialis wound infection associated with medicinal leech therapy. PMID- 8729212 TI - Helicobacter cinaedi bacteremia associated with localized pain but not with cellulitis. PMID- 8729213 TI - Extraintestinal Clostridium difficile infections. PMID- 8729214 TI - Rhodococcus equi mastoiditis in a patient with AIDS. PMID- 8729215 TI - Recurrent disseminated infection with Mycobacterium avium complex identified in tissues by molecular analysis. PMID- 8729216 TI - Mucormycotic osteolytic rib lesion presenting as subacute pleural effusion. PMID- 8729217 TI - Effect of clarithromycin on the pharmacokinetics of 2',3'-dideoxyinosine in patients who are seropositive for human immunodeficiency virus. PMID- 8729218 TI - Female genital blastomycosis: case report and review. PMID- 8729219 TI - Hepatitis E virus infection in Peru. PMID- 8729220 TI - Bartonella (Rochalimaea) henselae hepatosplenic infection occurring simultaneously in two siblings. PMID- 8729221 TI - Prosthetic valve endocarditis due to Staphylococcus saccharolyticus. PMID- 8729222 TI - Iatrogenically induced spondylodiskitis due to Mycobacterium xenopi in an immunocompetent patient. PMID- 8729223 TI - Acute pancreatitis associated with streptococcal toxic shock syndrome. PMID- 8729224 TI - Symptomatic vulvovaginitis due to fluconazole-resistant Candida albicans in a female who was not infected with human immunodeficiency virus. PMID- 8729225 TI - Serum levels of penicillin in basic trainees in the U.S. Army who received intramuscular penicillin G benzathine. PMID- 8729226 TI - Escherichia vulneris as a cause of intravenous catheter-related bacteremia. PMID- 8729227 TI - Aseptic meningitis secondary to carbamazepine therapy. PMID- 8729228 TI - Thoracic actinomycosis in a patient with pulmonary involvement caused by non Hodgkin's lymphoma. PMID- 8729229 TI - Isolated candidal epiglottitis: report of a fatal case. PMID- 8729230 TI - Flavimonas oryzihabitans sepsis in children. PMID- 8729231 TI - Suspected diphtheria in an Uzbek national: isolation of Corynebacterium pseudodiphtheriticum resulted in a false-positive presumptive diagnosis. PMID- 8729232 TI - Strongyloides stercoralis hyperinfection syndrome in a patient with AIDS: diagnosis by fluorescent microscopy. PMID- 8729233 TI - Strongyloides stercoralis first-stage larvae in the lungs of a patient with AIDS: primary localization or a noninvasive form of dissemination? PMID- 8729234 TI - Factors associated with the development of Pneumocystis carinii pneumonia. PMID- 8729235 TI - Salmonella aortitis. PMID- 8729236 TI - Toxoplasmosis localized to the bladder is diagnostic of human immunodeficiency virus infection: case report. PMID- 8729237 TI - Microbial detection. AB - There is a widespread need for commercial instrumentation for the rapid and inexpensive detection of microbial contamination of food, industrial waste water and clinical samples. A large number of detection methods have been developed utilizing the optical, electrochemical, biochemical and physical properties of microorganisms. The need for a device which can produce a rapid, accurate, sensitive, real-time analysis for clinical, industrial and environmental applications has led to considerable progress being achieved in recent years in the development of biosensors for microbial detection. This intense research has resulted in the commercialization of several instruments. Techniques used for the quantification of microorganisms are reviewed under the general categories of non bioelectrochemical and bioelectrochemical methods. PMID- 8729238 TI - Portable device for continuous fractionated blood sampling and continuous ex vivo blood glucose monitoring. AB - The objective of the study was to develop and evaluate a portable device for continuous fractionated blood sampling and continuous ex vivo monitoring of blood glucose. The inner lumen of a double lumen catheter (18 gauge x 45 mm) was placed in a peripheral vein and perfused with heparin solution (1.4 U min-1). The outer lumen was used to collect heparinized blood into 48 vacuum tubes at programmable sample volumes and time intervals (0.2-2 ml in 2.5-30 min). A sensor flow chamber with an internal volume of 1 mm3 incorporating a miniaturized thin-film amperometric glucose sensor was placed in the sampling line for continuous ex vivo blood glucose monitoring. Blood glucose and plasma insulin were measured during a frequently sampled intravenous glucose tolerance test (250 mg kg-1) and a subsequent oral glucose tolerance test (150 g) over 6 h in eight healthy volunteers (BMI 24.5 +/- 3.2 kg m-2). Additionally, in four experiments blood glucose was measured on-line using the glucose sensors. The overall correlation coefficients for whole blood glucose and plasma insulin between the manually drawn samples and the vacuum tubes were 0.73 and 0.87, respectively (p < 0.001). The miniaturized glucose sensor exhibited a linear measuring range of 25 mmol-1 glucose concentration and 95% response times of less than 30 s. Sensor readings and laboratory analyser results for the blood glucose measurement correlated between 0.93 and 0.98 (p < 0.001). In summary, continuous fractionated blood sampling and ex vivo blood glucose monitoring in ambulatory subjects is possible with a portable device. PMID- 8729239 TI - Bombardier beetle based biosensor. AB - A biosensor constructed by combining an oxygen electrode with the liquid ejected by bombardier beetles (Pheropsophus aequinoctialis) immobolized on a collagen membrane is developed for hydrogen peroxide. The sensor presents a linear range of 2.0 x 10(-4)-2.0 x 10(-3) M and a reproductibility of approximately 2%. It shows a better lifetime than similar ones employing purified catalase. The selectivity is good since other enzymes are not present in the natural source employed. PMID- 8729240 TI - Optimization of biosensing using grating couplers: immobilization on tantalum oxide waveguides. AB - This paper presents a comparative study of immobilization strategies for integrated optical grating couplers using tantalum oxide waveguides. As a model system the affinity reaction between protein G and human IgG was investigated. The receptors were coupled to the waveguide by adsorption, covalent attachment and avidin-biotin bridges after modifying the sensor surface by silanization introducing epoxy and amino groups. In addition, the results obtained by a monolayer coverage of the sensors with the receptors were compared with those using a carboxymethyl-dextran matrix allowing the immobilization of an increased amount of protein. The different coupling procedures were assessed by the system response due to the specific binding of human IgG to the sensing layer and are discussed with respect to their effectiveness, stability and the absence of non specific binding. PMID- 8729241 TI - Behaviour of surface acoustic wave interdigitated array electrode sensor in non aqueous solution and determination of blood plasma recalcification time. AB - A novel SAW-IDA sensor system was constructed for the first time by connecting the IDA electrodes in series with a SAW resonator. The frequency characteristics of the SAW-IDA sensor in non-aqueous solution were investigated. The effects of the parallel capacitance and cell constant were studied, and were calculated with the circuit network theory. These calculations provide guiding rules for design of the SAW-IDA sensor system. The SAW-IDA sensor was applied to determination of recalcification time and activated partial thromboplastin time of blood plasma. The sensor offers a new and effective way of studying clinical and laboratory haemostasis. PMID- 8729242 TI - Immobilization of proteins to lipid bilayers. AB - Phospholipid bilayers deposited on sensor surfaces are excellent substrates for immobilizing proteins via a molecular anchor. An integrated optics sensing device was used to accurately measure the binding kinetics of proteins thus anchored. By comparing the results with measurements using proteins from which the anchor had been enzymatically removed, it was shown that the anchor accounts for essentially all the irreversible binding. The insertion of the anchor into the lipid bilayer is a spontaneous process. This method of immobilization should be widely applicable to many soluble protein molecules, to which an anchor can be attached by routine methods of molecular engineering. PMID- 8729244 TI - Current awareness in biosensors & bioelectronics. PMID- 8729243 TI - Mediated amperometric determination of ammonia with a methanol dehydrogenase from Pseudomonas sp. AM-1 immobilized carbon paste electrode. AB - PQQ-dependent methanol dehydrogenase (EC 1.1.99.8) was immobilized onto the surface of a carbon paste electrode by the cross-linking method and the electrode was covered with a porous polycarbonate membrane. An electrocatalytic steady state current for the oxidation of methanol was observed using this electrode in the presence of phenazine methosulphate as an electron transfer mediator and NH4Cl as an activator for enzyme activity. The electrocatalytic current at +0.2 V vs. Ag/AgCl was analyzed. The properties of the electrode were examined for the determination of ammonia. The electrode responded linearly to ammonia in the range of 3 to 60 mM. The assay took 40 to 50 s. The electrode was used repeatedly at room temperature for 15 days and retained 50% of its initial activity. PMID- 8729245 TI - [Stable isotopes in nutrition research]. AB - During the last decades several new techniques have been developed for measuring the utilization of nutrients in humans. The development and potential use of techniques with stable isotopes are discussed. Examples of the different studies are presented for determining whole body aminoacid and protein dynamics like lipids and glucose metabolism in humans. The practical use of these diverse stable isotopically labeled biochemicals provide safe, ethical, noninvasive investigation of nutritional metabolism in humans. PMID- 8729246 TI - Agricultural and food policies. Some concerns regarding their nutritional relevance. AB - In recent times, nutrition analysts have been emphasizing the fact that the most critical nutritional unfulfilled need in underdeveloped countries is energy or quantity of food. This has prompted some leading food economist and agricultural policy makers to promote the extensive cultivation of high field inexpensive staples, including starchy roots and tubers. A typical example is cassava in many African countries and Indonesia. These foods not only have very little and poor quality protein, but also lack other essential nutrients. Interestingly, underdeveloped populations seem to select the nutritionally poorest staples under condition of extreme economic constraints, for example cassava or sorghum. But as their economic level improves they switch to nutritionally better staples, like wheat or rice. The people seem to be able to <> the need for quality rather than simply quantity as soon as an improved economic status allows them more choices. Some examples are given to support this contention. The present analysis emphasizes that the goals and objectives of nutrition scientists must be integrated with those of food and agricultural economists for the design of agricultural policies with increased relevance to human nutrition in a comprehensive way. Rather than proposing the cultivation of one inexpensive staple, these policies should consider a variety of complementary foods which would allow the people to chose diets providing sufficient quantities and balance of all essential nutrients in addition to energy. PMID- 8729247 TI - [Weight for height of Chilean school children according to age and puberal development]. AB - In a sample of 6371 school age children, aged 9 to 15 years, representative of four Regions of Chile (II,V,IX, and XIII) weight for stature was evaluated, acocording to age and puberal stages of Tanner. The averages and medians of weight for stature between 130 at 170 cm for men and between 130 and 163cm for females are presented. Higher weight for height whith age and puberal stages was found in both sexes, but differences were not always significants. A very low correlation between weight, and chronologic age and low between weight and puberal status was observed. This was reflected for higher differences of weight in the successive puberal stages than in successive ages. We conclude that the puberal development rahter than chronologic age, must be use for a better nutritional evaluation of adolescents. Nevertheless, other parameters with better distribution in puberal poblation, must be considered. PMID- 8729248 TI - [Effect of protein and carbohydrate preloads on food and energy intakes in preschool children with different nutritional status]. AB - Three preloads: high in simple carbohydrates (fruit juice), high in complex carbohydrates (banana) and high in protein (chicken meat), were tested in two groups of preschool children (56), aged 2 to 4 years from both genders. One group had a normal weight for height and the other high weight for height. The preload were given thirty minutes before lunch. The lunches consisted in a legume dish, apple and a milk dessert. Children were stimulated but not forced to eat and the amount of foods consumed was determined by differential weighing. The results showed that overweight children had a lower consumption at lunch that normal children when the high carbohydrate preloads were tested. High simple carbohydrate preload, 338,2 +/- 87.0 vs 350,9 +/- 75.9, high complex carbohydrate preload, 336.2 +/- 109.6 vs 375.4 +/- 89.4. High protein preload had not effect on the lunch consumption in the two groups of children studied. When an analysis by age of children were made, dividing the normal and overweight children in two groups, 24 to 36 months and 36 to 48 months, it was demonstrated that differences in lunch consumption between the normal and overweight children was derived essentailly from the results obtained in children of 24 to 36 months, high simple carbohydrate preload 282.3 +/- 88.9 g vs. 318.5 +/- 80.0 g and in the high complex carbohydrate preload, 262.6 +/- 98.9 g vs 364.5 +/- 116.3 g. When energy intake derived from food consumption was analyzed, the same tendency described for food consumption was demonstrated. The results presented show a different response of overweight children to the high carbohydrate preloads consumed before lunch, specially in children aged 24 to 36 months. This findings are important since they give an experimental information that can be utilized to find metabolic answers in relation to the onset of obesity in the childhood and to establish dietary recommendations in order to alleviate the risk of obesity in children. PMID- 8729249 TI - [Comparison between nutritional status of school children of low socioeconomic level from Santiago, Chile and Sao Paulo, Brazil]. AB - The objectives of this study were to determine and compare the anthropometric profiles of schoolchildren from low income household from Santiago, Chile and Sao Paulo, Brasil. A total of 1779 children from Santiago and 2210 from Sao Paulo were evaluated. Z-score distribution of the height/age (H/A), weight/age (W/A) and weight/height (W/H) indicators were utilized to assess the children. As a reference, the pattern recommended by WHO was used. The distribution of the H/A curves from both cities were deviated to the left. This deviation was more evident in Santiago. This situation points out the existence of a greater prevalence of growth retardation of the children from this city. In relation to the W/A indicator, both groups showed a similar distribution curves, Sao Paulo group however, had an increment of cases in the left extreme of the curve, under 2 <>. According to the W/H indicator the profile of both groups are different; one-Santiago-is deviated to the excess and Sao Paulo to the deficit. The differences observed in both groups of children studied indicated higher deficit of height, but lower current undernutrition in Santiago and increased wasting with conservated height in Sao Paulo. The observed situation suggests that the distinct patterns are consequences of socioeconomic factors at different stages of the growing process of the children and/or etnic differences of the population. PMID- 8729250 TI - Effects of dehulling and storage conditions on cooking requirements and chemical composition of soybeans. AB - Changes in cooking requirements and chemical composition of whole and dehulled soybeans, stored in 2 different environments [25 degrees C /75% R.H. (Environment 1) and 38 degrees C /90% R.H. (Environment 2)], were studied. Rate of water absorption and solid losses during cooking were higher for the dehulled soybeans at both storage conditions. However, cooking requirements to achieve the same degree of texture in the cotyledons were similar for whole and dehulled seeds. Cooking time increased with prolonged storage; the effect was more noticeable in samples stored under Environment 2. Samples kept for 6 months required almost twice as much cooking than control samples. Dehulled soybeans had a lower fiber content, relatively higher amounts of protein and fat, but similar amino acid compositions than whole soybeans. Cooking caused losses of carbohydrates and ash and, therefore, significantly increased levels of protein and fat reflected by losses of solids during soaking and cooking. Among the amino acids, only cysteine suffered substantial decrease as a result of cooking. Cooking and storage inactivated 99% and from 20-35% of the trypsin inhibitors, respectively; the latter effect was more accentuated in samples stored under Environment 2. PMID- 8729251 TI - Effects of dehulling, cooking and storage conditions on protein quality and digestibility of soybeans. AB - Soybeans were dehulled, stored under two environmental conditions [25 degrees C/75% R.H. (Env. 1) and 38 degrees C/90% R.H. (Env.2)], optimally cooked and assayed for trypsin inhibitor and protein quality with laboratory rats. Dehulling did not significantly affect protein quality (PER and NPR) and protein digestibility of raw and cooked soybeans. Raw soybeans diets were significantly poorer in protein quality and digestibility when compared with cooked counterparts. PER values of dehulled-cooked soybean diets decreased significantly (P<0.05) as seeds were stored for up to 3 months under either environment. There were no significant differences in PER values due to storage during the period from 3 to 6 months. PER values for whole-cooked soybean diets exhibited a significant decline only when stored for 6 months under Env. 2. PMID- 8729252 TI - The effect of soaking time on water absorption and solid losses of whole and dehulled soybeans. AB - Dehulled entire soybean cotyledons are required for the preparation of many interesting products. Removing hulls had a dramatic effect on the rate of water uptake during soaking. Maximum uptake was reached in only 3 hr compared to the 12 hr needed by whole beans. This saves time and reduces microbial growth during soaking. The amount of water absorbed by beans, with and without hulls, was similar once corrected for solid losses and surface water. Removal of fibrous shells during dehulling increased both the protein and oil contents by 2 %. Dehulled beans steadily lost solids during soaking. Losses were 8.6 % compared to 0.7 % for whole beans when maximum uptakes were first reached. Ways of avoiding or reducing these losses are discussed. PMID- 8729254 TI - [Food consumption surveillance]. AB - This paper analyzes the importance of food consumption surveillance; as food consumption is determined by multiple factors, the difficulties involved in this process are established. Some direct methods used for food consumption assessment and other indirect methods that can be used to infer it, are analyzed and discussed in terms of their advantages and disadvantages. Other methods based on the follow up of food basket cost and purchasing capacity of poor population wages in terms of some selected food commodities, are proposed. The importance of food consumption surveillance as an instrument for decision making is pointed out. PMID- 8729253 TI - [Effect of growing location on sensory properties and nutritional characteristics of black beans (Phaseolus vulgaris L.)]. AB - The present research consisted of an evaluation of five genotypes harvested from six growing locations. Variables of sensory properties, cooking quality and nutritional characteristics were determined. Genotype with longer cooking time was BV which also present hard shell. Those of shorter cooking time were FMB and PV. In Calera frosting during pod filling, drastically reduced cooking time, sensory properties and tannins. Taking this location off, the analysis show little effect of genotype or growing location in regard to determined properties. The genotypes with lower content of tannins were PV and BV. The content of lectins were in general low for all samples and the diferences between genotypes were not statistically significant (p<0.05) but they did for growing location. PMID- 8729255 TI - [Development of products for the elderly population: vitamin enriched pudding]. AB - An instant dessert powder, pudding type was developed to be consumed at lunch or dinner time. The dessert was designed to be prepared in skim milk and meets the nutritional needs of elderly people. The dessert contains modified starch, carragenine, vegetal fat, sacharose. Each serving has been enriched with 30% of the daily vitamin requirement advised for adults over 51 years old. The optimized process was carried out according to the Karlsruhe test. Each serving consists of 22g powder prepared in proportion of 18% in skim milk. The optimized dessert powder was controlled by means of physical, chemical and microbiological analyses. The sensory quality was determined in the ready to eat product and the acceptability level was measured in a group of people selected according to the characteristics of the target population. The dessert powder contains 1.1% protein, 5.2% fats, 89.76% carbohidrates, and provides 409 Kcal/100g. Both the sensory and microbiological quality were good and the level of acceptance reached 98%. PMID- 8729256 TI - [Manufacture of a freeze dried stew for mountaineers]. AB - The mountaineers use to eat at the evening dinner which ought to fulfil the following requirements: it should be nutritive and have an adequate composition, a small volume and should be light, keeping a good quality during a long period of time. Its preparation must be fast and easy, being well accepted by the group of mountaineers. A stew of common use in Chile based on meat and vegetables was elaborated, known as <>, a typical chilean stew. It was dehydrated by freeze-drying, a process based in the freezing and subsequent ice sublimation of the product, achieving a very high retention of both sensory and nutritional qualities. The final product corresponded to individual portions of 60g of Charquican which was vacuum-packed in a coextrude polyamide-polyethylene pack, which protects from the moisture and oxygen. It was used an outer packaging of plastic-coated aluminum foil, in order to protect him from the light. Controls of the optimized product indicate a 6,7% of humidity with an aw value of 0,26. The chemical composition is nutritionally adequate, with a 45.6% of carbohydrates, 5,0% of lipids and 29,5% of proteins. It presents a sensory quality with typical characteristics which ranged from good to very good. The microbiological quality meets the Standards of the Chilean Sanitary Regulation for Foods. The total plate count and the molds and yeasts count were below the proposed limits. The study of the Charquican s shelf life performed at environmental storage conditions (20-25 degrees C and 50-55 RH) showed that the microbiological quality is mantained without significant differences. The sensory quality presents a slight and gradual decrease until the average score fo 6,7 after 200 days storage. The evaluation with excursionists at the mountains, showed the excellent acceptability and the good reliability of the product. PMID- 8729257 TI - [Biochemical basis of enteral nutrition]. AB - Basic biochemical studies have demonstrated the products of protein intraluminal digestion are amino acids and peptides, and the those amino acids as well as small peptides are absorbed by independent mechanisms. The formers are absorbed by specific absorption systems mediated by carriers. The small peptides (di-and tripeptides) are absorbed intact from and may be intracelularlly hydrolised. Peptides with four or more residues are hydrolysed by peptidases located on the brush border of the intestinal villi and then absorbed as amino acids and/or small peptides. Such an absorption through a peptide mechanism is faster than the absorption of free amino acids. These are basic aspects that should direct the use of protein partial enzymatic hydrolisate in human nutrition. PMID- 8729258 TI - [Biological quality of a milk substitute for school children based on refined flour subjected to enzymatic hydrolysis]. AB - The biological quality of an experimental milk substitute based on a raw wheat flour subjected to enzymatic hydrolysis in comparison to a control product based on extruded flours and milk protein was studied in 35 <> female school age children. The girls were fed their customary diet during 2 consecutive 14 and 12 day periods, and randomized to the experimental and control products in a double blind crossover fashion. Apparent absorption of protein, energy, calcium and phosphorus was evaluated. Mean nitrogen intake from the experimental product was significantly lower (223 vs 244 mg/Kg/d p<0.0001). Absorbed nitrogen was also lower (187 vs 203 mg&kg/d p<0.0001). Energy intake and excretion were similar with both products; 96 and 95% of intake was absorbed for experimental and control products respectively. The mean calcium intake was significantly lower than the experimental product (39.7 vs 60.2 mg/kg/d p<0.0001). Absorbed calcium from the experimental product was 50% of control (20.7 vs. 39.5 mg/Kg/d p<0.0001). Phosphorus intake was also lower with the experimental product relative to control (22.0 vs 27.8 mg/Kg/d p<0.0001) and absorbed P was 13.1 vs 16.5 mg/Kg/d respectively. Both products were well tolerated. We conclude that the experimental product based on wheat flour does not differ significantly in protein and energy digestibility but calcium and phosphorus absorption and digestibility are significantly lower limiting its use in school feeding programs. PMID- 8729259 TI - [Risk factors of short stature in Chilean school children from rural areas of high social vulnerability]. AB - Growth retardation, a prevalent problem in Chile, was studied in rural countries of high social vulnerability. Risk factors were identified with a paired case control design. Domiciliary interviews of the parents of 161 pairs of children between 6-8 were conducted. Cases were children between -1 and -3z scores of height/age index and Controls were children between +/- 1/2z score. The higher significant relative risks analized with univariate methods (OR) were: large familiar group, limited domestic belongings, underweight during pregnancy, insufficient weight and height at birth, undernutrition and low stature of both parents. Conditional logistic regression model showed a high likelihood ratio of 98,20. The higher OR values were for father's stature (OR=4,98) mother's stature (OR=4,64), undernutrition (OR=4,53) and insufficient weight at birth (OR=3,23). These results indicate that small size in rural school age children of low socioeconomic level is determined by heredity, specially father's stature and adverse environmental factors, being undernutrition the main contribution. These results may became a significant subsidy to design [correction of desing] interventions to improve the nutritional status of children of low socioeconomic level. Thus contributing to develop all their physical growth potential. PMID- 8729260 TI - [Contribution of the school meal program--basic cycle--to the nutritional requirements of school children]. AB - The contribution of the meal given to Basic Cycle students of the Public Teaching Network in comparison to the daily recommendations of energy and nutrients was observed. The study was carried out in seven State schools in Piracicaba, SP. Weighing of food served in the school meal was used as a means for obtaining data during a 5 day period. The energy and nutrient contribution of the meal was calculated on an average <> basis, in comparison to the nutritional pattern defined by FAO/OMS/UNU (1985). Also, the nutritional quality of the meal was analyzed through the <>. The results showed that the meal offered to students contributes with approximately 30% of the total daily energy recommended and 100% of daily protein recommendations for the group of 7 to 8 year old students. Concerning vitamins, the average meal contribution was 30%, except for vitamin C. As to minerals calcium and iron the average composition of the meal observed to fulfill 30% of the total recommended for the group. The results of the study point the need for correcting failures in the nutritional content of meals since these are a major supplement in feeding the target group. PMID- 8729261 TI - [Agreement between animal and vegetable protein digestibility measured in vivo and in vitro and its effect on the chemical score]. AB - Protein digestibility is a key factor in the determination of protein quality using the chemical score. Since there are several methods available for determining protein digestibility the purpose of this study was to compare three methods in vitro (pH drop, pH stat and pepsin digestibility) and two methods in vivo (true and apparent digestibility in rats) in the determination of the protein digestibility of: casein, soy protein isolate, fish meal, black beans, corn meal and wheat flour. The results showed that in the case of highly digestible proteins all methods agreed very well. However, this agreement was much less apparent in the case of protein with digestibilities below 85%. As a result, the chemical score of these proteins varied substantially depending upon the method used to determine its digestibility. Thus, when the chemical score of the proteins analyzed was corrected by the true protein digestibility measured in rats, they ranked as: casein 83.56, soy 76.11, corn-beans mixtures (1:1) 58.14, fish meal 55.25, black beans 47.93, corn meal 46.06 and wheat flour 32.77. In contrast, when the chemical score of these proteins was corrected by the pepsin digestibility method, the lowest quality was assigned to fish meal. In summary, this results pointed out that for non conventional proteins of for known proteins which have been subjected to processing, protein digestibility should be measured in vivo. PMID- 8729262 TI - [Microbiological quality of street sold fruits in San Jose, Costa Rica]. AB - The sanitary quality of street sold fruits was analyzed during the period from march 1990 thru march 1993 in San Jose, Costa Rica. It looked for the presence of Salmonella spp. Shigella spp., Escherichia coli as well as fecal coliforms in natural refreshments, fruit salads and the fruits most frecuently expended on streets, either in slices as the pineapple (Ananas comosus), papaya (Carica papaya), non-ripe mangoe (Mangifera indica) and watermelon (Citrullus vulgaris) and those that can be eaten without peeling, like nances (Byrsonima crassifolia) and jocotes (Spondias purpurea). 25 samples of each fruit, 50 natural refreshments and 50 fruit salads were processed according to rinse solution method, and the bacteriological determination was based in the methodology described by Vanderzant & Splittstoesser and the Bacteriological Analytical Manual. In the same way, it was used the Most Probable Number for 5 tubes described in the Standar Methods of Water and Wastewater in orden to analyze 15 samples of ready to use water by the fruit hawker. The nutritional value was studied according to the food composition tables for Costa Rica, Latin America and USA. The results show that more than 30% of fruit samples, 70% of natural refreshments and 96% of fruit salad presented fecal coliforms. Same time, all of them present important contamination indexes with E. coli. Salmonella spp. and Shigella spp. were not isolated. The water analysis revelead that 53% contained fecal coliforms, probably due to the lack of hygiene in the utensils used to collect water. The nutritional evaluation shows that fruit portions (except watermelon) satisfy more than 100% of the diary recommendation of vitamin C (60 mg) and 4-7% of the recommended ingestion of dietetic fiber (30g). PMID- 8729263 TI - [Preservation and stability of corn tortillas at room temperature]. AB - Three treatments with chemical preservative (sodium propionate, potassium sorbate methylparaben and hydrogen peroxidemethyl paraben) were tested to delay microbial spoilage and extend shelf-life of corn tortillas at room temperature (25 degrees C). The treatment with the best results was selected for further studies using two types of packaging: Paper and high density polyethylene. Quality of corn tortillas during storage was assessed by measuring water content, microbial analysis (Total Plate Count, molds and yeast) and throguh sensory evaluation. Results were analyzed by covariance analysis and slope contrast between packaging materials at p<0.05. Spoilage of tortilla without preservative occurred within 24 hours due to a large number of gram negative bacteria, molds and yeasts, which were responsible for offensive odors. Only the combination of hydrogen peroxide methyl paraben had a significant effect on retarding bacterial yeast spoilage. In addition, hydrogen peroxide residues could not [correction of no] be chemically detected after 2 days of storage. Results from this study show that tortilla can be kept for up to six days at room temperature with acceptable sensory properties with proper preservative treatment and packaging. PMID- 8729264 TI - [Cara-de-rama (Dioscorea bulbifera, L) fried processing]. AB - The present study examined the influence of density of the aerial bulbs in the variables: fat absorption, fry time and yield of the cara-de-rama chips and french fries, as well as organoleptic analysis of fried products. The results showed that there was no differences between the bulbs densities, in the variables examined, except for the moisture of cara-de-rama chips. In the organoleptical analysis, the samples did not show significant differences in relation to similar potato products. PMID- 8729265 TI - [Evaluation of biological fish silage in broiler chicken]. AB - Biological fish silage was produced from the mixture of three low price fish species. After mixing it with molasse, papaya and pineapple wastes, and Lactobacillus plantarum incoculation, the product was stored for seven days, dehydrated and added to several diets of formulated poultry feed as a substitute of fish meal. Physical-chemical and microbiological analyses were done to the raw materials and final products. Biological tests of acceptability indicated that chicken preferred diets up to 50% of fish silage. Chicken's weigh increased and food consumption tests, in addition to the sensory evaluation tests in the chickens white meat, showed the advantages of inclusion of 5-20% fish silage in the diets of experimental animals. PMID- 8729266 TI - Evaluation of bush and vine black beans for physical, chemical and nutritional characteristics. AB - The present study was undertaken to learn if there are physical, chemical and nutritional differences between vine and bush type of beans. Four samples of black color beans (Phaseolus vulgaris) of the vine type, and four of the bush type were collected from farmers in the same growing area. The samples were analyzed for some physical properties including, 100 seed weight, size distribution percent seed coat, water absorption, cooking time, and of solids on cooking waters. Vine type beans had larger 100-seed weights, larger sized beans, thicker seed coats, and lower of solids in the cooking water than bush type beans. Rate of water absorption was different. The chemical characterization included proximate analysis and fiber fractionation. Vine type beans had, on the average, less ether extract and protein than bush type. No differences were found in fiber fractions, although there was a higher variability in the vine types. Protein quality and protein digestibility when fed as the single protein source, were similar on the average, with more variability in the vine types. Both types, efficiently supplemented maize proteins and the protein digestibility was higher than when fed alone. In general there were no large differences, except in some physical measurements, between vine and bush type beans, with the former showing greater nutritional variability which could be useful in selection programs, if such variability is confirmed. PMID- 8729267 TI - [Chemical composition, dietary fiber and mineral content of frequently consumed foods in northwest Mexico]. AB - Nutrient composition in foods is very important specially in evaluation of nutritional status in populations. In this study the proximate composition, dietary fiber (DF) and mineral content of 15 frequently consumed foods in Northwest Mexico were determined. The procedures used were AOAC (1984) official methods, chemical-enzymatic method for DF and atomic absorption spectrophometry for minerals. Foods were grouped into cereals, legumes, meat and dairy products, fat was the most variable component in all foods (0,41 to 21,1 g/100 g). Fired beans (Phaseolus vulgaris: variedad pinto) had the highest DF content (9,21 g/100g); as is basis). Sodium among the minerales was also highly variable mainly due to the addition of salt during preparation of foods, except in corn tortillas were salt is not added. In contrast wheat flour tortillas had the highest sodium content of the foods analysed. Fresh white cheese had the highest calcium content (563 mg/100g). The meat group had the highest content of Fe and Zn (2,4-5,4 and 4,2-5,4 mg/100 g respectively). This study has provided information with current analytical techniques of important foods in northwest Mexico that will contribute to food composition tables in Latin America. PMID- 8729268 TI - [Comparison of analized and calculated energy, fat, protein, dietary fiber, iron and zinc values in diets from different socioeconomic levels in northern Mexico]. AB - Traditional methods of dietary assessment such as measuring nutrient intake with 24 h dietary recalls, food frequency questionnaires and multiple-day food records, depend upon the use of data base systems to estimate nutrient data. Certain problems exist with the data from these sources. For Mexico, the most serious one is that in many of the systems certain nutrient data is lacking and many of the traditional foods are not included. The objective of this study was to analyze regional diets for protein, fat, dietary fiber, iron and zinc and compare these values with those estimated from two different data bases, ALIM 10.000 which includes regional dishes and foods and Nutritionist III, which includes data from Handbook 8. Energy values were calculated using reported values. The results showed that the bases produced comparable values to those analyzed for energy, protein, fat, however for micronutrients the data bases generally overestimate the analyzed values. The results of this study emphasize the need to update data bases with new product information, re-examine certain values on basis of newer methods, and to include data for ethnic foods. PMID- 8729269 TI - A comparison of eutherian and marsupial optic chiasms: a brief review. AB - There are four fiber components in the mammalian optic chiasm. Two crossed and two uncrossed components, one of each from each eye. The two crossed components meet at the midline, forming coarse interdigitating bundles in some species, and extremely fine crossing bundles in others. The two components from one eye mingle in the optic nerve. Then in some species they separate, before reaching the chiasm, but in others they do not separate until they reach the midline of the chiasm. Once past the chiasm, the two components that form the tract reunite and one more, the crossed and the uncrossed fibers mingle. The review considers that difference between the marsupial and the eutherian mammals that have been studied, and looks at the developmental changes that are likely to underlie these different types of chiasmatic organization. PMID- 8729270 TI - When neurobiology and genetics meet: the study of visual system development in Drosophila melanogaster. AB - The visual system of Drosophila melanogaster is an exceptionally well suited model to study the molecular and genetic mechanisms underlying inter- and intracellular signalling during nervous system development. This review highlights some of the recent advances in this field, that show not only the power of the combined neurobiological and genetical approaches in shedding new light into long standing questions regarding nervous system development but also uncover a striking evolutionary conservation, at the molecular level, of the basic developmental pathways and signal transduction cascades underlying metazoan ontogenesis. PMID- 8729271 TI - Localization and mapping of the main gene responsible for fast development in Drosophila melanogaster. AB - Drosophila melanogaster Oregon-R lab stock was subjected to mass selection during 682 and 459 generations for fast and slow developmental time, respectively. This paper reports the experiments that located the major gene for fast developmental time on the third chromosome by use of an enzymatic system as well as body markers. The recombination mapping using means of eclosion placed this fast gene at 3.46.55. PMID- 8729272 TI - [Isoenzymatic characterization of the hexokinase in the development of Plebeia droryana (Hymenoptera, Apidae) and its relation with flight activity]. AB - Hexokinase enzyme was studied by means of starch-agarose gel electrophoresis in Plebeia droryana. Three anodal bands with enzyme activity were observed during the development: hexokinase-1 (HK-1), the intensity of which increases from larvae to adult, probably related with energy supply to thoracic flight muscles and, therefore, with flight activity; hexokinase-2 (HK-2), that reaches maximum intensity in clear brown eyed pupae and hexokinase-3 (HK-3), the intensity of which reaches maximum peak in winged pupae and is not observed in the adult phase. This isozyme should have important functions in the bee metamorphosis. PMID- 8729273 TI - Effects of juvenile hormones I, II and III, in single and fractionated dosage in Melipona bees. AB - Applications of Juvenile Hormones JH I, II and III in single or fractionated doses to larvae of Melipona compressipes, Melipona quadrifasciata, Melipona rufiventris and Melipona scutellaris at the L3 and beginning of cocoon-spinning phase (pre-defecating larvae - LPD) activated feminizing genes, inducing differentiation of female larvae into queens. The technique of fractionated treatment proved to be highly efficient in Melipona species for producing of queens. It is difficult to obtain 100% queens with single dosage of JH in some species which implies in a mechanism of JH degradation by specific esterases. One hundred per cent of queen production occurs only when an adequate amount of Juvenile Hormone is administered within the period that is critical for caste determination. The threshold dosage of JH I for Melipona compressipes (below which it is not possible to obtain 100% queens) was 0.1mu JH I 4 applications of 0.25mu g each. For Melipona quadrifaciata the threshold dose for obtaining the highest proportion of queens was 0.2mu g JH I in 4 applications of 0.05mu g each. For Melipona scutellaris 0.025mu g JH I/mj 1 in single dosage induced 100% of queens. No 100% production of queens in Melipona Rufiventris treated larvae were obtained; the highest proportion was 86% of queens with 0.2mg g JH I/mu 1. In order to obtain expression of the feminizing genes in Melipona species, JH I was the most efficient followed by JH III. Each species respond differentialy to JH dosage, and this dosage must be tested before use. PMID- 8729274 TI - [The effect of hemolymph inoculation on the mechanism of defense of Biomphalaria tenagophila (Orbigny, 1835)]. AB - The resistance of B. tenagophila snails to infection by Schistosoma mansoni was studied. These snails had been previously inoculated with hemolymph talsen from other snails infected by S. mansoni of SJ strain or by other trematodes. The findings suggest that two types--cellular and humoral--are responsible for the resistance to infection by S. mansoni. The cellular response occurred when the snails were inoculated with hemolymph from snails infected by S. mansoni, B. tenagophila inoculate with hemolymph from snails infected by furcocercariae without eye-spot, also showed resistance to the infection by S. mansoni. In this latter case, the degenerated sporocysts were not enveloped by amebocitary reaction, leading to the conclusion that humoral factors could be involved. PMID- 8729275 TI - [Relationship between the pathogenicity of Schistosoma mansoni in mice and the susceptibility of the vector mollusk. III. Mortality, body and viscera weight]. AB - The relationship between the development of hepatomegaly, splenomegaly, body weight and mortality rate and the degree of susceptibility of Biomphalaria glabrata and B. tenagophila at which infective cercariae developed was studied. The study utilized Swiss mice, SPF, as definitive hosts and populations of snails genetically selected for character susceptibility as intermediate hosts. Low body weight and smaller viscera were observed in infected mice with cercariae originating from snails that showed a high degree of susceptibility. The higher susceptibility of molluscs infected with S. mansoni corresponded to a lower survival of the infected mice. These results lead to the conclusion that the higher degree of adaptation of the parasite to its intermediate hosts, evidenced by the high indexes of susceptibility, leads to different behaviour on the part of this parasite in its definitive host. PMID- 8729276 TI - Studies on mosquitoes (Diptera: Culicidae) and anthropic environment. 10- Survey of adult behaviour of Culex nigripalpus and other species of Culex (Culex) in south-eastern Brazil. AB - A survey of adult behaviour of Culex (Culex) species was carried out from August 1992 through December 1993 in a human modified (anthropic) environment in the Ribeira Valley, S. Paulo State, Brazil. Culex nigripalpus dominated the catches at several sites and it's tendency to increase in the anthropic environment became quite clear. Nevertheless no high level of synanthropy was demonstrated. So it seems that the mosquito may have a restricted role in natural arbovirus cycles. Nonetheless, Cx. nigripalpus must be considered a potential vector of arboviruses, especially St. Louis encephalitis virus outside dwellings. PMID- 8729277 TI - [Evaluation of the excess of cases of tuberculosis due to HIV/AIDS infection: a preliminary assay]. AB - An exercise for the estimation of the percentage risk of cases of tuberculosis attributable to co-infection HIV/AIDS, using the following formula, is propounded: RA%=p[m2r(hR-h)] + (1-p)[m3r (hR-h)]/p[m1+m2r (hR+1-h)] + (1-p)[m3r (hR+1-h)] x 100 where: p= proportion of BK infected, r= risk of tuberculosis infection, h= proportion of persons infected with HIV, m1= breakdown rate of endogenous tuberculosis, m2= breakdown rate of exogenous tuberculosis, m3= breakdown rate of primary tuberculosis, R= relative risk of morbidity among persons infected with HIV. PMID- 8729278 TI - [The use of stratification and logistic regression model in the data analysis of case-control studies]. AB - Data of a case-control study of esophageal cancer were used as an example of the use of multivariate analysis with stratification and logistic regression. Eighty five cases and 292 controls were classified according to sex, age and smoking and drinking habits. The point estimates of the odds ratios were similar, and the techniques were considered complementary. PMID- 8729279 TI - [Evaluation and control of the microbiological quality of hands in foodhandlers]. AB - Microbiological analyses of workers' hands were made for the common indicators, including aerobic mesophilic plate counts (APC), as well as the common food pathogens. Opportunities were observed for cross-contamination of roast beef by workers' hands during slicing operations. Workers' hands showed APC counts of up to 10(7) CFU/hand and the presence of S. aureus and C. perfringens. Salmonella spp were not isolated from hands. These results show that handling of these foods by such workers would be a risk in transmitting pathogenic microorganisms to the foods and is apparent that it is necessary for these workers to take care of personal hygiene. Decimal reductions obtained in the microbiological counts after washing and antisepsis of workers' hands were at 2,6 logs cycles and still demonstrated the importance of this practice in food services by the fact that pathogens such as S. aureus and C. perfringens were inhibited or killed. PMID- 8729280 TI - [Vitamin A deficiency in children of rural areas of the semi-arid region in Bahia]. AB - The distribution and magnitude of vitamin A deficiency and dietary consumption of 161 children at 6 to 72 months of age in rural zones in Cansancao-Bahia-Brazil were evaluated. The serum retinol levels were measured by the spectrophotometric method (Bassey-Lowry modified by Araujo and Flores). The serum retinol average was found to be distributed homogeneously throughout the different age groups. Inadequate serum retinol levels (<20,0 mu g/dl) were detected in 44.7% of the children, which characterized the deficiency as constituting a public health problem. The serum retinol levels showed no statistically significant association as between the sex and age of the children; however the children of less than 24 months showed a higher prevalence of inadequate serum retinol levels. The main available source of vitamin A for these children was represented by carotenoids, especially beta-carotene. Foods regarded as being rich in vitamin A were consumed by all age groups. The greatest diversification of consumption of foodstuffs with moderate and low vitamin A content was observed in the group of children of from 24 to 72 months of age, through this was no guarantee of adequate serum retinol levels in this group however. PMID- 8729281 TI - [The effect of the use of milk fortified with iron and vitamin C on hemoglobin levels and nutritional status of children under 2]. AB - The impact of the use of fortified powdered whole milk (9 mg of iron and 65 mg of vitamin C/100 g of milk) on the hemoglobin levels of children under 2 years of age was evaluated, over a period of 6 months, in 107 children enrolled in municipal Day Care Centers (DCC) and in 228 seen at a Basic Health Care Unit (BHCU). Before the beginning of the intervention, 66.4% of the children in the DCC and 72.8% of those seen at the BHCU had hemoglobin levels under 11.0 g/dl. After 6 months of fortified milk intake, these percentages fell to 20.6% and 18.0% respectively. The average hemoglobin before the intervention was 10.3 g/dl in the DCC and 10.5 in the BHCU. After 6 months these increased to 11.6 g/dl in the populations studied. Concerning the nutritional condition, evaluated according to Gomez's criteria, 57% of the DCC children presented an improvement, 41.1% showed changes and only 1.9% became worse. In the BHCU, 11.4% presented better condition, 70.6% remained the same and 18% worsened, which demonstrated differences in response regarding improvement of nutritional condition, when fortified milk was used in closed and open environments. The authors conclude that the utilization of enriched foods is an excellent alternative in the treatment of iron deficiency in populations of children under 2 years of age. PMID- 8729283 TI - [Exploratory research: a methodological procedure for the study of human factors in the field of Public Health]. AB - Exploratory research is proposed as a methodological procedure for a qualitative approach. The main purpose of this procedure is that of helping the researcher to develop a measurement tool which reflects reality. Both exploratory research conceptions, the traditional and the new one are discussed. The application of this methodological procedure to the study of human factors is recommended and the implementation stages are presented. Some applications for research in public health are suggested. PMID- 8729282 TI - [Assessment of strategies of science and technology management: a case study]. AB - The Health Department of the State of S. Paulo has implemented a management strategy on research and development (R&D) for its research institutes which comprises three different items: 1) project registration, 2) support and 3) promotion of R&D activities. The present study explores the ability of this R&D management system to provide information and evaluates the performance of the third item which has been responsible for the assignment of research grants to professionals whose projects are selected through a peer review. The assessment of the grant's distribution was conducted by comparing the mean scientific production recorded in literature data bases for grant holders and equal an number of controls matched by institute of origin. The ability of the system to produce information was studied by the application of Cluster Analysis. The results show that grant holders have a scientific production significantly higher than their controls. Concerning information production, it is shown that proper raw data processing may render suggestions for decision making and that the institutes may be grouped in clusters of those that adopt similar management strategies. PMID- 8729284 TI - [Advances and challenges in health economics]. AB - Health economics is a specialized field of economic science that applies the economic perspective to the fields of health, the medical-industrial complex and health services. A brief review of the evolution of this speciality by subject, as well as the level achieved assessed in terms of generation, diffusion, reproduction and application of its specialized knowledge, is presented. PMID- 8729285 TI - [Centro Universitario de Atencion del Adolescente (C.U.A.A.). Catedra e Pediatria y Neonatologia. Universidad Nacional de Cordoba]. PMID- 8729286 TI - [Age of menarche. Secondary sex characteristics. Interrelation. Secular trend]. AB - Our purpose was to determine the mean age of menarche in young females of our population. To relate menarche with maturation stages of secondary sex character. To observe if the secular trend persist after the first observation. Consistently two temporal observations spaciated for 10 years were performed. Two hundred and eighty three healthy women who belonged to a higher middle-level and attending a high school from Cordoba National University were studied. Such women were in an age range between 11.0 to 18.0 years. Anthropometric and biological maturation examination were undertaken (Tanner). The mean age of onset of secondary sex characters was determined in a longitudinal follow-up group (54 girls). The mean age of menarche in the overall sample was determined by the retrospective method and in the subsample of 54 girls longitudinally followed in the 70-80 decade was determined by the prospective method in which the menarche was correlated with stages of mamary development (B) an pubic hair (PH). The mean age of menarche in 146 young females with a follow-up from 1.981 through 1.990 was determined. In the overall sample the mean of age of menarche was of 12.59 years +/- 1.17; in the subsample (54 girls) of 12.75 years +/- 1.03. In this subsample 70.37% of them has the menses in the B3 stage, 22.22% in B4, 5,5% in B2 and 1.85% in B5. 74.07% PH3, 18.51% in PH4, 0.52% in PH2 and none in PH5. In the sample of 146 young females with a follow-up between 1981 and 1990 the mean age of menarche was 12.36 years. If this result is compared with that of the 54 girls longitudinally followed in the prior decade and with the overall sample, a difference of 0.39 years and 0.23 years respectively is proved. This difference favoring an earlier age of menarche would show: 1 degree) That in our environment the secular trend is maintained, the age advances 4 months every 10 years and 2 degree) That the longitudinal prospective method is more reliable than the retrospective one. PMID- 8729287 TI - [Anthropometric evolution of height, weight and total body water pre and post menarche]. AB - The aim of the survey was to assess the anthropometric evolution of height, weight and total body water 1 year prior and until 1 year after the menarche in a higher middle level scholar adolescents group from a double-time secondary school belonging to Cordoba National University. A longitudinal study of 48 adolescents was performed. The control was made every six months and at the time of menarche, Twuenty anthropometric variables were relieved from which height and weight were used, the total body water (TBW) was determined by Mellits and Cheek equation and biological maturation was assessed according to Tanner criteria. Height and weight, and total body water increase pre- and post-menarche was determined, with +/- 1SD, as well as the average of studied variables +/- 1 SD. Height and weight increase was of 7.813 cm (+/-1.61) and 7.488 Kg(+/-2.3) respectively 12 months prior menarche, with a major speed in the first semester. After menarche, in the following months, height as much as weight increase, more the former than the latter, but there is less increase than in the year previously to menarche. The increase in the 12 posterior months has an average of 3.48 cm(+/-1.18) for height and 2,524 Kg(+/-1.90) for weight, with a major speed in the first semester. Likewise, the total body water decrease as an index of the body fat and is inversely correlated with the increase of weight in fat. We conclude that these results are perfectly correlated with the studied biological phenomenon, according to patterns described by other authors in other countries with different racial features. The onset of menarche is a good parameter to predict, rather approximately, without using another estimates, the final height in a girl. PMID- 8729288 TI - [Body composition at menarche. Estimation of total body weight, total body water, lean and fat body weight]. AB - Our aim was to confirm in our environment what has been observed and described by other writers about the importance of achieving a "critical body weight'' and an adequate "fat percentage'' -on the basis of the calculation of total body water- for the initiation and development of pubertal events. This study included 92 girls, healthy, well nourished, belonging to upper middle class from a high school of The National University of Cordoba. The longitudinal method of control was used every 6 months and at the precise moment of menarche. Out of 20 antropometrical variables observed height, weight and height, TBW as percentage of body weight, lean body and fat weight, fat percentage and skin folds ppercentiles for each girl at menarche. A regression between fat percentage and skin folds was done. Percentiles 5 to 95 of fat percentage in relation to body water percentage were estimated. At menarche the average for the different variables are: Heigth 155.6 cm +/- 0.469; Weight 45.8 Kg +/- 0,5; TBW 25.216 lit. +/- 0.318; lean body weigth 35.02 Kg (S.D.2.98); fat weigth 10.86 Kg (S. D. 3.17). The addition of skin folds was correlated fat percentage, thus, an equation was obtained for the average calculation of such percentage %F= 12.16 + (0.313 x fold addition). The minium percentage for the onset of menstrual cycles is 17.3% and corresponds to percentile 10. However, there is a 5% of girls who start to menstruate with a 15.5% of fat and none of them is below that value. The reasons mentioned above suggest that is necessary to obtain a "critical body weigth'' as well as a "fat percentage'' minimum for the onset and maintenance of menstrual cycles, among our girls, similar o what has been obtained by doctor Frisch. PMID- 8729289 TI - [Pubertal development in males]. AB - The purpose of this study was to investigate the characteristics of the male puberal development in our population. Two hundred and ninety seven health males belonging to a higher middle social economic level who attended a secondary School for Cordoba National University were studied. Such males are encompassed in an age range between 11.0 and 18.0 years. Clinical, anthropometric and biological maturation examination were undertaken according Tanner criteria. The mean age of onset in secondary sex character with their corresponding SD (s), E.E.x. and C.V., the correlation between them by Pearson's correlation coefficient ??r'' and the duration of the complete puberal process were determined. The first sign of male puberal development is the testicular enlargement, appearing G2 at a mean age of, 12,43 years (SD 0,7475), G3 12,74 years (SD0,8873) G4 13,75 years (SD0,9488), and G5 15,28 years (SD 1,0834). The public hair appears at a mean age of PH2 12,66 years, PH3 13,31 years PH4 14,55 years, and PH5 15,38 years. At 17,5 years 100% of the males have reached their complete development. The existing correlation between variables, comparing genital development with pubic hair and viceversa, pubic hair with genital development was of 0,92 and 0,91 respectively. The complete puberal process in males encompass from 2 years 2 months to 3 years 6 months, mean of 2 years 11 months. The male genital development (G2) beging 8 months after the girls started their mammary development (B2) and the male pubic hair (PH2) appears one year later than in the girls. PMID- 8729290 TI - [Sexual behavior in adolescents and young subjects]. AB - The purpose of this paper was to obtain information about sexual behavior of university students. To achieve this goal we implemented a survey which was answered anonymously by 438 students, 123 women and 315 men. They were divided into three age groups. Between women, sexual activity grew with age, but between men, on the contrary, the highest percentage was verified in the youngest age group. The age at which sexual activity was started decreased in one year each four years of age and is two years ahead in boys. Reasons stated for this decision by women were based on values and love, while pleasure and opportunity were appointed by men. Both men and women of all ages showed a high number of sexual partners. Sexual transmitted diseases were acknowledged by a high number of students. This results clearly show the necessity of an educational programme including sexual advisory and sexual transmitted diseases prevention in all health care actions designed for university students. PMID- 8729291 TI - [Adolescents and young subjects facing contraception, pregnancy and abortion]. AB - The objective aimed was to obtain information about the sexual behavior of adolescents and youths attending university as regards contraception, pregnancy and abortion and the knowledge of the Papanicolau test as a preventive, method for malignant diseases. An anonymous survey which was especially designed for this investigation was implemented with 438 students - 123 women and 315 men - divided into three groups according to age; between 17 - 20, 21 - 24 and more than 25 years-old. Taking into account the sexually active population. 70% of the men between 17-20 use contraceptive methods (condoms 98,6%) and so do 100% of the women of the same age (menstrual cycle control method 71,4%, contraceptive pills 14,2%, or her partner uses condom 21,4%). In the group of youths between 21-24 years old, 76,8% of the men and 82,7% of the women use contraceptive methods: 70% of the men use condoms and 30% contraceptive methods used by their patner, whereas 58,3% of the women use the menstrual cycle control method, 25% take pills and 8,3% use DIU. In the group of older youths 71,4% of the men use contraception methods (66,6% condoms) and 51,14% of the women (100% the menstrual cycle control method) and 25% add other methods to this one. Only 20,68% of the women stated having had pregnancies (all of the between 21 - 24). As there is the same porcentage of abortion in this age groups, it can be presumed that all the pregnancies ended in abortion. This contradicts the position stated regarding abortion due to the fact that 66% of the population questioned in the survey was against abortion and defended the right to live, whereas 23% was for abortion mainly in the case of rapes and/or maternal diseases. A small porcentage was for abortion in the case of pregnancies which may disturb their life projects. As regards the Papanicolau test, all the groups have a good knowledge about it, but only women over 25 years old practice it responsably. PMID- 8729292 TI - [Primary prevention actions in AIDS and sexually transmitted diseases in adolescents]. AB - Considering primary prevention of ETS a priority in Adolescent Health care, we decided to investigate knowledge and attitudes of 205 students in relation to these pathologies. Results were used to organize an educational proposal based on groupal training of ??Youth Health Promotors''. It is also shown the necessity of joint work between family and school in order to think and revise together with the young people involved, themes and values, in order to make a coherent, creative and holistic approach to human sexuality. PMID- 8729293 TI - Experimental autoimmune myocarditis: a suitable model to study neuroimmune crosstalk. AB - This review regards the main functional characteristics of hearts subjected to an autoimmune response, focusing especially on the role of T lymphocytes and autoantibodies in the development of cardiac dysfunction. Evidence of a strong association in the onset and time-course of immune response and cardiac dysfunction is presented and the results are viewed comparatively with myocarditis models induced by heart, parasite or virus inoculation. Cardiac damage is evaluated regarding various aspects, namely histologic, immunologic, biochemical, pharmacologic, physiologic. Finally, the model, for its characteristics of resulting from an autoimmune response against the heart with functional consequences, has proved its usefulness to study neuroimmune interaction, mainly the immune to nervous direction, as autoantibodies and T cell derived factors have a role in cardiac failure. PMID- 8729294 TI - [XIIIth improving course in rhythmology. La Grande Motte, France, June 29-30, 1995]. PMID- 8729295 TI - [Double responses]. AB - Double response is a rare electrocardiographic phenomenon requiring two atrioventricular conduction pathways with very different electrophysiological properties. Double ventricular responses are the usual manifestation: an atrial depolarisation (spontaneous or provoked, anticipated or not) is followed by a first ventricular response dependent on an accessory pathway or a rapid nodal pathway and then a second response resulting from sufficiently delayed transmission through a nodal pathway for the ventricles to have recovered their excitability when the second wave of activation reaches them. A simple curiosity when isolated and occurring under unusual conditions, particularly during electrophysiological investigation of the Wolff-Parkinson-White syndrome, the double response may initiate symptomatic non-reentrant junctional tachycardia when associated with nodal duality and repeating from atria in sinus rhythm. The functional incapacity and resistance to antiarrhythmic therapy may require referral for ablation of the slow pathway. PMID- 8729296 TI - [Ventricular tachycardia in the normal heart]. AB - Idiopathic ventricular tachycardia is not a single clinical entity. Several electrophysiological mechanisms are responsible for at least two typical electrocardiographical pictures. The tachycardia with right bundle branch block morphology and inferior axis is the most common type and originates in the right ventricular outflow tract. It has to be differentiated from right ventricular dysplasia, and also from tachycardia in association with mitral valve prolapse. It is responsive to calcium antagonists, and can be treated with radiofrequency ablation. The other tachycardia (left bundle branch block morphology and right axis) originates in the posterior left ventricular septum and has typically sharp potentials preceding the ventricular electrogram in the site where ablation is successful. Both types can be associated with different types of cardiomyopathy, and therefore further investigation might be useful. PMID- 8729297 TI - [Dynamics of ventricular repolarisation]. AB - The clinical value of assessing the QT interval is obvious as it is a marker of ventricular depolarisation and repolarisation and it allows identification of clinical situations carrying a high risk of ventricular fibrillation and sudden death. Until the last few years, analysis of ventricular repolarisation was based on analysis of conventional surface electrocardiography. Technical difficulties explain the limits of our knowledge of the dynamics of ventricular repolarisation. This situation is beginning to change rapidly by computerised analysis of Holter monitoring, opening up a particularly complex and important field of research. The duration of the QT interval depends on different factors, especially changes in electrolyte balance, effects of certain drugs, and changes in heart rate and autonomic nervous system tone. The difficulty resides in selecting the pertinent data in order to study separately the effects of heart rate and those of the autonomic nervous system. The initial results show that this analysis provides important information for diagnosis and probably prognosis, on the status of the myocardium and the action of the autonomous nervous system. They require confirmation and validation on larger series of patients with different pathological conditions. PMID- 8729298 TI - [Anti-arrhythmic therapy and cardiac failure]. AB - In cardiac failure, continuous ambulatory electrocardiographic recording for 24 hours (Holter system) enables detection of 60 to 80% of complex ventricular arrhythmias, 15 to 40% of atrial arrhythmias and sudden death accounts for about 40% of fatalities but its causes are multiple and sometimes unrelated to arrhythmias. Abnormalities of cardiac structure, metabolic and neuro-hormonal changes and some drug therapies are implicated in the genesis of these arrhythmias, the management of which is discussed in two different situations with respect to the functional incapacity: in paucisymptomatic ventricular arrhythmias in patients with cardiac failure, class I antiarrhythmics and d sotalol should be avoided and betablockers prescribed with caution; the indications of amiodarone have not yet been determined. When the arrhythmia is symptomatic (sustained ventricular tachycardia or ventricular fibrillation), class I antiarrhythmics are not effective enough in the prevention of sudden death; betablockers and amiodarone may give good results but should be compared with implantable defibrillators in the future. The multiplicity and complexity of the mechanisms of arrhythmias in cardiac failure, and the inadequate results obtained with classical antiarrhythmics necessitate the development of new antiarrhythmics based on blockade of non-selective channels probably activated in cardiac failure by the stretching of myocardial fibres. PMID- 8729299 TI - [The esophageal approach in rhythmology]. AB - The possibility of detecting the electrical activity of the heart from the oesophageus has been recognised for nearly a century. On the other hand, transesophageal pacing has only been really developped in the last fifteen years, which explains the recent interest for this technique in clinical practice. Easily put into practice, but not always well tolerated, the oesophageal approach has many uses in rhythmology. The principal diagnostic applications are in unlabelled tachycardias whether with narrow or wide QRS complexes, the evaluation of the Wolff-Parkinson-White syndrome, the study of sinus node function or nodal conduction. The therapeutic applications are dominated by the reduction of supraventricular tachycardias especially atrial flutter, with a success rate similar to that of endocavitary stimulation. The facility of realisation, especially at the patient's bedside, without need for fluoroscopie control, makes it a useful tool in emergencies, especially if the endocavitary approach cannot be used. The only reserve is the painful character of pacing in some patients. PMID- 8729300 TI - [Non-pharmacological treatment of atrial fibrillation]. AB - Although often denigrated in recent years, antiarrhythmic drugs frequently enable control of atrial fibrillation in patients who are symptomatic or at high risk of complications. In patients resistant to therapy, a number of non-pharmacological options have become available. The choice is no longer limited to his bundle ablation, a method previously at the point of progress but which has gradually given way to more sophisticated and less definitively destructive techniques. Modulation of atrioventricular conduction is currently under development, the feasibility of which has not yet been completely evaluated, but whichmay provide a less radical alternative in selected patients. Restoration of sinus rhythm by electrical cardioversion has benefitted from the advances of cardiac defibrillation in general. The theoretical information which its use implies has improved its efficacy, especially in the case of endocavitary electrical shock. This method can also be employed with automatic implantable defibrillators but using specific algorithms. Atrial pacing is credited with a preventive efficacy but this has never been definitely demonstrated. Research is under way in this domain. Future hopes are also centered on atrial surgery, especially the operation described by Cox which is generally accepted to be effective and which may be adapted medically using endocavitary radiofrequency current. Most of these methods are insufficiently experimented but they should be considered reasonably in particularly symptomatic patients duely informed of the nature of the treatments which are proposed. PMID- 8729301 TI - [New indications for cardiac pacing]. AB - New indications have recently appeared for cardiac pacing with haemodynamic and antiarrhythmic objectives without any symptomatic bradycardia. The best documented indication, though relatively rare, is stimulation of obstructive hypertrophic cardiomyopathy; initially reserved for cases with favorable results of an acute haemodynamic test, it is now used in other cases without this criterion; hypertrophic cardiomyopathy without permanent obstruction, atrial fibrillation or left bundle branch block. The improvement observed during follow up is always greater as a real remodeling of the myocardium seems to occur with ventricular dilatation and/or septal thinning. However, the position of the atrial, and above all, of the ventricular pacing catheters is critical as is regulation of the pacemaker which should allow complete ventricular capture with an AV delay allowing good filling. The follow-up of these patients must therefore be regular and the effects on longevity are unknown. DDD pacing has also been proposed in dilated cardiomyopathy. The results are contradictory and only very selected cases with left bundle branch block and long PR interval seem justified with, again, optimisation of the pacing sites with high septal or biventricular stimulation. Recurrent atrial tachycardia, special algorithms preventing extrasystoles have been tried with variable results. In cases with inter-atrial block, atrial resynchronisation by bi-atrial stimulation has been assessed with promising results but many technical problems remain unsolved. PMID- 8729302 TI - [Transient entrainment of tachycardia]. AB - Transient entrainment of tachycardia by pacing at a faster rate corresponds to its acceleration with return to sinus rhythm either when the pacing is interrupted or when the pacing rate is slowed. It conforms to strict criteria detectable either on the surface electrocardiogramme (constant fusion, progressive fusion, change of morphology before termination of a tachycardia) or on selective endocavitary recording when entrainment is hidden. The phenomenon described by Waldo in 1977 in atrial flutter is found in all reentrant tachycardias. It is currently being studied in ventricular tachycardia. The electrophysiological concept is useful because it facilitates the understanding of the mode of termination of tachycardia by stimulation and the localisation of zones for destruction during ablation. PMID- 8729303 TI - ["Within" the law? or "outside" of the law? Apropos of the law on clinical research]. PMID- 8729304 TI - [Evaluation of professional practice and health care organization in anesthesia and intensive care: a strategic approach]. PMID- 8729305 TI - [Preoperative hemodilution by erythrocytapheresis with homologous blood saving in total hip arthroplasty]. AB - OBJECTIVES: To compare three techniques for decreasing homologous blood requirements in total hip arthroplasty (THA), including preoperative autologous donation (PAD), preoperative acute normovolaemic haemodilution with erythrocytapheresis (erythro) and intraoperative normovolaemic haemodilution (haemo). STUDY DESIGN: Prospective clinical trial. PATIENTS: The study included 45 patients scheduled for THA, under general anaesthesia and operated on by the same surgeon. The patients were allocated into three groups of 15 each. METHODS: Blood loss was assessed, during surgical procedure, by the weight of sponges and, the amount of blood collected in the suction bottles during and after surgery. The haemoglobin concentration was measured at the time of preoperative assessement (d-30), just prior to surgery (d-1), in the recovery room (d+3h), and 1, 3, and 8 days later (d8). The transfusion end-point in the three groups was to obtain a haemoglobin concentration of 100 g.L-1 from d+3h until d8. Every pack of red blood cells transfused was weighed and its haematocrit assessed to determine the accurate volume of red blood cells. RESULTS: In the three groups haemoglobin concentration was similar from d+3h until d8. In the PAD group, no patient required homologous blood transfusion. There was no significant difference between the two other groups in the mean volume of homologous red blood cells required (308 +/- 197 mL in erythro group and 331 +/- 202 mL in the haemo group, respectively). The intraoperative blood loss was significantly higher (P = 0.001) in the erythro group: 914 +/- 305 mL vs 665 +/- 263 in the PAD group and 512 +/- 146 mL in the haemo group, respectively. There was an inverse correlation between haematocrit at d-1 and intraoperative bleeding (r = -0.7) (P = 0.0001). The distribution of the points was fitted as an exponential curve. CONCLUSIONS: In THA, PAD is obviously the best technique to avoid homologous blood transfusion. However, when PAD is not feasible, removal of blood prior to surgery does not decrease requirements of homologous blood, as intraoperative blood loss is higher. Our results strongly question the use of major haemodilution during a surgical procedure exposing a major blood loss. PMID- 8729306 TI - [Transfusion of erythrocyte concentrates. An evaluation in anesthesia and intensive care]. AB - OBJECTIVE: To assess the relevance of perioperative packed red blood cell (PRBC) transfusion practice at the University Hospital of Nice, compared with information from the consensus conference on red blood cell transfusion, held by the French Society of Anaesthesia and Intensive Care (SFAR) and the National Agency for the Development of Medical Evaluation (ANDEM) in December 1993. STUDY DESIGN: Retrospective case series analysis. PATIENTS: The study included 240 medical files of surgical patients, transfused in 1994 with PRBC, obtained by drawing of lots following a methodology recommended by ANDEM. METHOD: A reference list according to the statement of the consensus conference was designed for the various surgical specialities and the ICU in which PRBC had been transfused. It included the clinical and laboratory criteria which justified the transfusion, as well as the various categories of PRBC (phenotyped, cytomegalovirus negative, leukocyte-depleted, etc). Autotransfused PRBC were also considered. The data collected from the medical files of the 240 patients were compared with the reference list. RESULTS: In 84.6% of patients (203/240), the PRBC transfusion had been decided with reference either only to a haematocrit level below 0.27 or a level between 0.27 and 0.30 associated with clinical evidence of bad tolerance of blood loss, according to the reference list. A lack of compliance with the reference list occurred in 15.4% of patients (37/240), who had been transfused without any reference to a biological criterion. Another non compliance existed in 50% of patients (12/24) transfused with phenotyped PRBC and in 35.3% (6/17) of those transfused with leucocyte-depleted PRBC. An autotransfusion with PRBC had been carried out in 30.4% of patients (75/240). DISCUSSION: These deviations of transfusion practice from the consensus conference statement, which were more pronounced with phenotyped and leucocyte-depleted PRCB than conventional PRBC, resulted in the edition of a report, with an analysis of the causes of deviations and recommendations for all doctors of our institution prescribing blood transfusions. Another evaluation, extended also to the medical specialities of our hospital and including all blood derivates is planned for 1996. PMID- 8729307 TI - [Severe burnt patients: hemodynamic state, oxygen transport and consumption, plasma cytokines]. AB - OBJECTIVE: To assess the haemodynamic and oxymetric variations measured by a pulmonary artery catheter and to correlate them with the variations of the circulating cytokines during the initial intensive care phase of severely burned patients. STUDY DESIGN: Prospective study covering an 18-month period. PATIENTS: Thirteen successive patients, aged over 12 years, without significant medical history, with a thermal burn affecting more than 50 percent of their total body surface area and admitted to our centre during the first six postburn hours. METHODS: The haemodynamic and oxymetric profile was investigated by inserting a blood flow-directed balloon-tipped pulmonary artery fiberoptical catheter. All patients were treated according to the protocol previously used in our centre. Blood samples were drawn on admission, every 12 hours post-injury until the 2nd day, then on the 3rd and 5th days. Cytokines were analyzed by Elisa method. Haemodynamic and oxymetric measurements were achieved simultaneously with the biological samples during the first 5 postburn days. The analysis of variance (ANOVA) with the Duncan test was utilized for multiple comparisons between continuous variables. RESULTS: (mean +/- SEM): The patients were 32 +/- 3 years old and had a burn surface of 72 +/- 4%. After a short hypovolemic shock period lasting a 12 hours, a hyperdynamic shock occured which increased until the 5th day, with an increased cardiac index (6.9 +/- 0.4 at h120 vs 2.9 +/- 0.3 L.min 1.m-2 at h6, P < 0.05), increased oxygen transport and consumption (respectively 880 +/- 77 at h72 vs 543 +/- 58 mL.min-1 at h12, P < 0.05 and, 203 +/- 15 at h72 vs 129 +/- 25 mL.min-1 at h6, P < 0.05) and markedly decreased systemic vascular resistances (1,002 +/- 118 at h36 vs 2,330 +/- 328 dyn.s.cm-5.m2 at h6, P < 0.05). Circulating cytokines were not clearly modified except for interleukine-6 which reached early striking peaks (16,858 +/- 10,330 at h24 and 15,406 +/- 6,509 pg.mL-1 at h36) simultaneously with the decrease in systemic vascular resistances. CONCLUSIONS: During the first post-injury week, critically burned patients develop a specific hyperdynamic circulatory status during which interleukine-6 could be a mainfactor decreasing systemic arterial resistances. PMID- 8729308 TI - [Prevention of increase of blood pressure and intracranial pressure during endotracheal intubation in neurosurgery: esmolol versus lidocaine]. AB - OBJECTIVES: To compare the preventive effects of esmolol and lidocaine on the increase in mean arterial pressure (MAP) and intracranial pressure (ICP) during endotracheal intubation in neurosurgery. STUDY DESIGN: Comparative, randomised, double-blind study. PATIENTS: Twenty-two patients, physical status ASA I or II, undergoing neurosurgery, and randomised into two groups (esmolol group and lidocaine group). METHODS: After induction of anaesthesia with thiopentone, vecuronium, fentanyl and isoflurane, one group received iv esmolol 1.5 mg.kg-1 and the other iv lidocaine 1.5 mg.kg-1, 130 sec before endotracheal intubation. The MAP measured with a radial catheter, the ICP obtained with a lumbar subarachnoid catheter and the cerebral perfusion pressure (CPP, calculated from MAP and ICP) were assessed before induction of anaesthesia, before esmolol or lidocaine injection, and before intubation, during the maximal change in MAP, as well as 2 and 5 minutes after intubation. RESULTS: The time course of MAP, ICP and CCP were similar throughout the study in the two groups, with a significant decrease (P < 0.05) of the CPP from 92 +/- 12 to 62 +/- 8 mmHg after esmolol, and from 96 +/- 12 to 68 +/- 15 mmHg after lidocaine. Following intubation, CPP increased significantly (P < 0.05) to 99 +/- 23 mmHg after esmolol and to 99 +/- 17 mmHg after lidocaine. The ICP increased also significantly (P < 0.05) after intubation from 11 +/- 6 to 17 +/- 10 mmHg in the esmolol group, and from 10 +/- 6 to 16 +/- 9 mmHg in the lidocaine group. CONCLUSIONS: Esmolol or lidocaine as an iv bolus of 1.5 mg.kg-1 before laryngoscopy and intubation do not completely prevent the increase in MAP and ICP. PMID- 8729309 TI - [Comparative study of buprenorphine and its combination to ketoprofen or propacetamol for postoperative analgesia in urologic surgery]. AB - OBJECTIVE: To compare the analgesic effect of subcutaneous buprenorphine alone and in combination with propacetamol and ketoprofen following urologic surgery. STUDY DESIGN: Open randomized clinical trial. PATIENTS: Sixty ASA II/III patients undergoing urologic surgery. METHODS: The patients were randomized into three groups to receive either buprenorphine (0.3 mg subcutaneous) on demand (group 1, n = 20), or a combination of buprenorphine (0.3 mg)-propacetamol (2 g)-ketoprofen (100 mg) by intravenous route over 2 hours followed by an infusion of propacetamol (2 g) and ketoprofen (100 mg) at a constant rate over. The remaining 22 hours (group 2, n = 20), or the same loading dose as in group 2 prolonged by a continuous infusion of buprenorphine (0.3 mg), propacetamol (2 g) and ketoprofen (100 mg) over the same period (group 3, n = 20). Visual analogue scale pain scores (0-10) were assessed every hour during the 24 hours of the study. When the VAS score exceeded 5, an additional dose of 0.3 mg of buprenorphine was administered. RESULTS: Groups were similar for age, surgery, anaesthesia and initial pain levels. Compared to group 1, the onset of analgesia was earlier in groups 2 and 3 at the 1st hour (P < 0.05); the level of analgesia was lower at the 3rd hour (P < 0.05). The maintenance of this analgesia level required constant buprenorphine administration. Buprenorphine requirements were decreased to 56% and 37% in groups 2 and 3 respectively, compared to group 1 (P < 0.05). Incidence of nausea and vomiting was lowered to 15% in group 3 (P < 0.05). CONCLUSION: A combination of buprenorphine, propacetamol and ketoprofen provides effective postoperative analgesia with a low incidence of nausea and vomiting and decreased requirements of buprenorphine. PMID- 8729310 TI - [Prospective evaluation of a nuclear magnetic resonance score in the screening of malignant hyperthermia susceptibility. Initial results]. AB - A prospective study for the noninvasive diagnosis of malignant hyperthermia (MH) susceptibility was conducted in 30 patients using 31P magnetic resonance spectroscopy (MRS). A score of MRS muscle abnormalities was determined before the in vitro contracture test. The patients were classified as MH susceptible or MH negative, according to an algorithm of MRS score values. Twenty-three patients were correctly classified using the MRS test, five had inclusive MRS score values and two patients were false-positive. There were no false-negative patients. These preliminary results suggest that the MRS test could be useful as a possible noninvasive diagnostic test in MH susceptibility. PMID- 8729311 TI - [Numerical expression of results]. PMID- 8729312 TI - [Quality assessment in anesthesia]. AB - Quality assessment (assurance/improvement) is the set of methods used to measure and improve the delivered care and the department's performance against pre established criteria or standards. The four stages of the self-maintained quality assessment cycle are: problem identification, problem analysis, problem correction and evaluation of corrective actions. Quality assessment is a measurable entity for which it is necessary to define and calibrate measurement parameters (indicators) from available data gathered from the hospital anaesthesia environment. Problem identification comes from the accumulation of indicators. There are four types of quality indicators: structure, process, outcome and sentinel indicators. The latter signal a quality defect, are independent of outcomes, are easier to analyse by statistical methods and closely related to processes and main targets of quality improvement. The three types of methods to analyse the problems (indicators) are: peer review, quantitative methods and risks management techniques. Peer review is performed by qualified anaesthesiologists. To improve its validity, the review process should be explicited and conclusions based on standards of practice and literature references. The quantitative methods are statistical analyses applied to the collected data and presented in a graphic format (histogram, Pareto diagram, control charts). The risks management techniques include: a) critical incident analysis establishing an objective relationship between a 'critical' event and the associated human behaviours; b) system accident analysis, based on the fact that accidents continue to occur despite safety systems and sophisticated technologies, checks of all the process components leading to the impredictable outcome and not just the human factors; c) cause-effect diagrams facilitate the problem analysis in reducing its causes to four fundamental components (persons, regulations, equipment, process). Definition and implementation of corrective measures, based on the findings of the two previous stages, are the third step of the evaluation cycle. The Hawthorne effect is an outcome improvement, before the implementation of any corrective actions. Verification of the implemented actions is the final and mandatory step closing the evaluation cycle. PMID- 8729313 TI - [A survey of operating and recovery room ventilators and monitoring equipment as well as their maintenance]. AB - OBJECTIVE: To survey the monitoring and anaesthetic devices available in a sample of French hospitals and the modalities of their maintenance. STUDY DESIGN AND METHOD: Survey carried out in 21 private and public hospitals (including eight university hospitals), affilated to the French Association for Research in Anaesthesia and Intensive Care (ARAR). RESULTS: More than 84% of all operating theaters were equipped with at least an ECG, a SpO2, and a non invasive blood pressure monitor. The non equipped anaesthesia sites were mainly delivery rooms, plaster cast rooms and X-ray rooms. The figure of recovery room beds was, generally speaking in accordance with the French recommendations. Maintenance policy was not systematic and not budgeted in 72% of the surveyed centres. CONCLUSION: The equipment in most anesthetic sites has significantly increased, however maintenance policy remains still heterogenous. PMID- 8729314 TI - [Collection of cases of death from anesthesia based on death certificates. A feasibility study. Report to the General Direction of health]. PMID- 8729315 TI - [History of respiratory intensive care with reference to first aid for nearly drowned persons]. AB - In addition to anecdotal techniques originating from Galen's reasoning, the methods for the treatment of nearly-drowned persons from the XVIIIth century to the first quater of the XIXth century, are surprising as they still include the artificial internal ventilation techniques of today. Nevertheless for more than one century, these techniques were abandoned. External manual methods generating a forced expiration were preferred until 1958, when Peter Safar and coworkers demonstrated the value of mouth-to-mouth ventilation in comparison with manual methods. This eclipse finds its origin mainly in Leroy d'Etiolles's 'alarming' dissertation produced in 1827, underlining the pleuro-pulmonary risks of endotracheal insufflation and recommending the external manual methods, considered as being easy to use by everybody, without risk, and however improperly given as being efficient. As a result, the European philantropic societies abandoned bellows and positive pressure ventilation in their official resuscitation scheme for the nearly-drowned and asphyxiated. PMID- 8729316 TI - [Secured freeze-dried plasma. Its use in war conditions]. PMID- 8729317 TI - [Immunologic therapy of blood coagulation disorders with circulating anticoagulants]. PMID- 8729318 TI - [Tracheotomy: to push the intubation tube down...or how to avoid technical problems]. PMID- 8729319 TI - [Percutaneous dilatation tracheotomy: importance of the choice of cannula]. PMID- 8729320 TI - [Citation of works published in AFAR]. PMID- 8729321 TI - [XIV Consensus Conference on Intensive care and emergency medicine. Management of status epilepticus (children-adults). 23 June 1995, Le Kremlin-Bicetre]. PMID- 8729322 TI - [What are the advances in the treatment of acute kidney failure?]. AB - Mortality due to acute renal failure has remained high over the last three decades despite a better understanding of the pathophysiologic mechanics involved and advances in the management of critically ill patients. This paradoxical situation raises many questions concerning the criteria used to assess treatment and disease severity as well as the effectiveness of certain recently proned "advances". In the 1980s, much progress was made in preventing and/or limiting the extent of acute renal failure. The use of very early and vigorous fluid administration associated with alkaline diuresis prevents traumatic rhabdomyolysis; saline hydration before and after radiocontrast administration protects against the acute decrease in renal function in high-risk patients; maintenance of an adequate intravascular volume and of blood pressure helps prevent acute renal failure. After the onset of acute failure, low-dose dopamine can increase urine output, whereas dobutamine improves creatinine clearance but there is no evidence that increasing urine output lowers morbidity or mortality. New techniques have been proposed for acute renal replacement therapy, but except for use of bicarbonate dialysis and biocompatible membranes, none have been shown to be superior. It has been claimed, on the basis of uncontrolled or retrospective studies, that continuous hemofiltration or hemodiafiltration could have a beneficial effect on survival and/or the course of infectious complications. However, convincing evidence of this beneficial effect is lacking since these techniques are incompatible with clinically pertinent removal of proinflammatory cytokines. The type of renal support may have no effect on outcome. Can further progress be expected in the future? Antagonists of NO receptors and growth factors have a protective effect on renal function in animal models, results which may be of clinical relevance. Their clinical potential should be evaluated in prospective randomized trials involving patients where severity of illness is assessed at inclusion using a multiparametric model combining a severity score and relevant prognostic factors. PMID- 8729323 TI - [Traumatic spinal complications of cervical arthrosis]. AB - OBJECTIVES: Cervical spondylotic myelopathy is usually a chronic and progressive disease. In a few cases, however, dramatic tetraplegia sometimes occurs after even minor injury. METHODS: We report seven patients (6 males, 1 female, aged from 41 to 63), who suffered from acute myelopathy after an injury. RESULTS: In 5 cases, the injury revealed the cervical spondylotic myelopathy. A hyperextension of the cervical spine was found in 5 cases. The injury was severe in only one case, but there was no bony abnormalities, except arthrosis. The most common cause was a fall. In contrast, severe tetraplegia was found in 4 cases. Spontaneous, but incomplete, recuperation occurred in 4 patients. In all 7 cases, CT scan and MRI showed congenital cervical stenosis associated with cervical spondylosis. The level of disco-osteophytic changes was mainly in C4, C5, C6. On T2-weighted spin-echo image, an increased signal intensity was present in the cord of 3 patients, but was not correlated with the severity of the symptoms, nor with improvement. Surgical treatment was performed in 6 cases: 3 laminectomies, 3 anterior or antero-lateral approaches. Improvement after operative decompression was observed in all but one case, even when the motor or sensory deficit persists for more than one year. Motricity of the inferior limbs improved better than the other deficits, perhaps because of the location of non-reversible lesions in the spinal gray matter. CONCLUSION: The acute medullary syndrome of cervical spondylotic myelopathy is serious and can cause major handicaps. This complication justifies a preventive surgical attitude when medullar signs are moderate, and cervical imagery shows a spondylotic compression of the cord with congenital stenosis. PMID- 8729324 TI - [Management of HELLP syndrome before 32 weeks of amenorrhea. 22 cases]. AB - OBJECTIVES: Assess expression and management of HELLP syndrome (hemolysis, elevated liver enzymes, low platelet count) occurring before 32 weeks gestation. METHODS: Among 50 patients presenting HELLP syndrome from 1990 to 1994, 22 (44%) who developed the syndrome before 32 weeks gestation were evaluated retrospectively. RESULTS: Most of the patients were primiparous and HELLP syndrome recurred in 2 during a second gestation before 32 weeks gestation. Only three cases began during the post partum period. All patients had severe pre eclampsia before discovery of the HELLP syndrome. Episodes of eclampsia also occurred in 6. The most frequent clinical manifestation was epigastric pain. Ten patients had acute severe renal failure. The 3 post partum patients had severe complications (eclampsia, renal failure, subcapsular hepatic hematoma). Obstetrical intervention was required in all cases. Cesarean section was performed within 48 hours of diagnosis. Pregnancy had to be terminated in 3 cases between 24 and 29 weeks gestation. There was one fetal death in utero and one during the neonatal period. Seventeen live infants were delivered. In the group of 11 infants born after 30 weeks gestation, only 1 had hyalin membrane disease which developed in all those born before 30 weeks, including 2 with broncho pulmonary dysplasia. CONCLUSION: Based on the physiological mechanisms involved in HELLP syndrome, criteria for obstetrical extraction and the possibilities for conservative management in very premature pregnancies, we propose a management protocol for HELLP syndrome developing before 32 weeks gestation. Corticosteroid therapy may be given for 48 hours in cases without maternal or fetal complications in order to accelerate fetal maturation before extraction. PMID- 8729325 TI - [Calcification of the pectineal ligament. An usual radiological imaging in pediatrics]. AB - OBJECTIVES: Since its first description several years ago in the elderly subject, calcification of the Cooper ligament is often considered as a rare anecdotal phenomenon. We present the radiographic presentation which is often not well known. METHODS: Two experienced radiologists examined independently a series of 100 consecutive X-rays of the pelvis performed in a population of elderly subjects (mean age 84.4 +/- 9.2), looking for calcification of the Cooper ligament. RESULTS: Calcification of the Cooper ligament was observed on 7 of the 100 images of the pelvis. There was no significant difference in age, sex, or reason for ordering the examination between subjects with calcification and those without calcifications. Arterial calcifications were observed at an equal frequency. The usual aspect of the Cooper ligament was a fine opaque line following along the upper border of the iliopectinate crest. Oblique views visualized the calcification better. The line of calcification was either continuous of fragmented and was bilateral in all 7 cases. CONCLUSION: These images are apparently of no pathological significance. Clinicians should however be aware of the radiographic presentation in order to avoid confusion with visceral calcification or periosteal deposits. PMID- 8729326 TI - [A wine pitcher, cause of lead poisoning]. AB - A 35-year-old male nurse hospitalized for recurrent episodes of intolerable abdominal pain was found to have non-hemolytic anemia and saturnism with blood lead level reaching 500 micrograms/l. Search for the source of the lead led to the discovery of an earthenware jug purchased in a Corsica craftsware shop. This jug had been used to hold the patient's daily consumption of wine (estimated at one-half to three-quarters liters per day) in the refrigerator. On the basis of lead release measurements using the method described in the EEC directive 84/500 (maximum tolerated limit for crockery = 4 mg lead/litter contents), it was found that the jug released 216 mg/l and estimated that the patient had ingested 0.8 to 1 mg of soluble lead daily for the past 8 years. The patient was successfully treated with chelator agents. This case of lead poisoning caused by a single earthenware wine jug confirms the need for rigourous governmental directives to control the production and distribution of varnished earthenware. PMID- 8729327 TI - [Post-traumatic thrombosis of the portal vein]. AB - Blunt trauma to the abdomen is an exceptional cause of portal vein thrombosis. To our knowledge, 8 cases have been reported in the literature. When thrombosis of the portal vein occurs, a complete search for all the known main causes must be carried out before entertaining this diagnosis. Other causes may be cirrhosis, tumors and inflammation of the abdomen, coagulation disorders and hematologic diseases including latent myeloproliferative syndrome. We report a case in a 25 year-old man with an uneventful past history who presented with thrombosis of the portal vein after a violent blunt trauma which occurred during a rugby play. In this young man, none of the other potential causes was found, in particular bone marrow culture on medium with low growth-factor concentration allowed us to eliminate a latent myeloproliferative syndrome. The only triggering factor remaining was the recent abdominal trauma. After an 18-month follow-up, no other element has been observed which could have caused thrombosis of the portal vein. PMID- 8729328 TI - [Bone structure and mechanical resistance of the bone tissue]. AB - The strength of bone depends on bone structure, both in terms of global geometry of the bone and the microscopic pattern of the trabecular network. Modifications in the trabecular organization, often related to the aging process, include altered anisotropy, perforations, loss of connectivity and microfractures. The microscopic architecture of trabecular bone can be studied on histomorphometric slices, the use of stereologic techniques on these slices may better reflect the changes occurring in 3 dimensions. High-resolution tomodensitometry and 3-D magnetic resonance imaging are under further evaluation. An evaluation of the fractal geometry of the trabecular network, performed on the vertebrae, calcaneus, or ulna on standard radiographs has given promising results. Bone strength is determined by resistance tests on bone tissue samples. The relationship between the applied force and bone deformation is used to determine a stress-strain curve and resistance parameters. The mechanical behavior of a given bone can be modelized by a Finite Element Analysis, the computerized model being able to predict the reaction of this bone. Ultra-sound analysis does not directly measure bone density, nor bone architecture, but would reflect both these properties. Velocity and attenuation of ultra-sounds, measured at the calcaneus or patella, are useful in distinguishing between controls and osteoporosis cases, and may have predictive value. PMID- 8729329 TI - [Blood transfusions in cancer patients]. AB - In this review, we develop four topics on the relationship between blood transfusion and cancer. First, the rationale for not allowing blood donations from patients with infiltrating tumors is presented. Second, the different possibilities for autotransfusions in cancer patients are discussed. Predeposited autotransfusions are rarely possible in these patients, in addition to the high cost. The usefulness of another method, intraoperative autologous transfusion with blood saved from the surgical field, is not well established. Our third topic concerns the effect of transfusion on cancer induction. In some cases, the risk of cancer is higher after allogenic transfusion resulting from a mechanism involving alterations of the immune function. Finally, the relationship between transfusion and cancer recurrence is controversial. In spite of numerous studies attempting to elucidate this relationship, no final conclusion can be drawn at the present time. What is sure, is that patients requiring blood transfusion have a higher risk of recurrence than patients who do not need transfusion. PMID- 8729330 TI - [Chylothorax of unusual origin: post-radiation mediastino-pulmonary fibrosis]. PMID- 8729331 TI - [Has troponin I its role in the early screening of myocardial necrosis?]. PMID- 8729332 TI - [Akathisia induced by low doses of neuroleptics after a pallidal infarction]. PMID- 8729333 TI - [Annular ectasia of the aorta in a patient with acquired cutis laxa]. PMID- 8729334 TI - [Apropos of the diagnosis of preimplantation in transgenic mice]. PMID- 8729335 TI - [The peripharyngeal space. Anatomy and normal imaging]. AB - Sectional anatomy and dissection allow topographic study of the peripharyngeal region. Anatomic study has been performed by projection on the skull base of the limiting fascia and by 6 mm thick anatomic slices in the three fundamental planes. Three spaces are identified: retropharyngeal space represents a slipping structure upon the cervical spine. The prestylian space is mostly formed by an adipose column along the pharynx and by the medial part of the parotid gland. The retrosylian space includes the internal jugular and carotid vessels, the four last cranial nerves and the cervical part of the laterovertebral sympathetic trunk. CT an MR imaging were performed on ten patients with cervical or cephalic diseases distant from peripharyngeal regions. CT imaging is only useful in axial and frontal views for the prestylian space. MR imaging is the best method to visualize the three components of the peripharyngeal region. Confronting sectional imaging with anatomic slices allows better recognizing of limits and contents of the different parts of peripharyngeal regions. PMID- 8729336 TI - [Ultrasonographic aspects of hepatic metastases of thyroid medullary cancers]. AB - PURPOSE: The aim of this study was to describe the different ultrasonic features of hepatic metastases of medullary thyroid carcinoma and to point out two particular patterns. MATERIAL AND METHOD: A retrospective review of the sonographic examinations of 35 patients with hepatic metastases from medullary thyroid carcinoma was performed. The metastases were classified relatively to their size and sonographic appearance. RESULTS: Four ultrasonic types of metastases were described: Type I: small (< 3 cm) hyperechoic nodules with the same features as typical hepatic hemangioma (54%); Type II: markedly hyperechoic micronodules often associated with acoustic shadowing (40%); Type III: hyperechoic nodules of more than 3 cm in diameter (14%) corresponding to advanced intrahepatic disease; Type IV: hypoechoic or in target form nodules (28%) rarely isolated (8%), mostly associated with additional hyperechoic lesions (20%). Thirty two out of 35 patients had hyperechoic nodules; in 22 patients the HM were of a single type, mainly type I (n = 12) and in 13 patients different types of nodules were present. These sonographic features were correlated with the histologic characteristics of medullary thyroid carcinoma. No precise relationship could be establish between the histologic form of MTC and the US features. CONCLUSION: The authors underline the possible mistake between the metastases of type I and the hemangioma and the characteristic appearance of metastases of type II. PMID- 8729337 TI - [Interventional MRI. Value of low field strength and very fast 3D steady state imaging sequences for the guidance of a "non" ferromagnetic needle]. AB - PURPOSE: The excellent tissue differenciation provided by MRI in the three directions and the lack of ionizing radiations make it ideal for interventional procedures. Many problems must still be solved: the access to the patient, the size of the artefact produced by interventional instruments and the acquisition time. In this study, we evaluated the influence of field strength for standard imaging sequences on the MRI artefact of a non ferro-magnetic biopsy needle. Then, we developed very fast sequences on a opened 0.1 T dedicated magnet to test in vitro and ex vivo the potential ability of this system in the guidance of the needle. MATERIAL AND METHODS: The biopsy needle was a new stainless steel needle made of 49% Nickel. The needle artefact's size measurements were made at 0.1 T, 0.5 T and 1 T on a bicompartmental phantom (one compartment with a long T1 and another with a long T2), with standard gradient echo (400/12/90 degrees) and spin echo (500/25) sequences. At 0.1 T, we optimized very fast steady state 3D FAST (T1 weighted) and 3D CE-FAST (T2 weighted) sequences to reduce the acquisition time, preserving good image contrasts for a field of view reaching 38 mm and 48 x 64 or 24 x 32 matrix. RESULTS: The larger needle artefact observed on gradient echo images varied from 3.6 mm at 0.1 T to 8.6 mm at 1 T. The shortest acquisition time for 4 contiguous slices of 2mm with a 1.2 mm/pixel resolution and a 24 x 32 matrix was 1.5 s for the 3D FAST (16/9/65 degrees) sequence and 3 s for the 3D CE-FAST (29/22/65 degrees) sequence. We realized a complete MRI guided abdominal puncture on a cat cadaver with 4 series of 15 s 3D FAST images (16/9/65 degrees, 4 slices, 5 excitations, 1.2 mm/pixel, FOV = 77 mm). Besides the cat positioning and the image reconstruction time, the whole puncture lasted 1 min (4 x 15 s). CONCLUSION: Low field MRI (0.1 T) combined with very fast 3D steady state sequences is adapted for the real time guidance of biopsy needles. PMID- 8729338 TI - [Ultrasound-guided transparietal thoracic puncture biopsy]. AB - Transthoracic needle biopsy is a reliable method for the diagnosis of thoracic lesions, specially when they are peripleral. AIM: The main objective was to evaluate the results of US guidance for thoracic needle biopsy in 159 cases. RESULT: Overall sensitivity was 83% as tissue was obtained in 132/159 cases. In 24 cases, 2 or 3 biopsies were necessary. In 17 cases, analysis was made on cytology only with a 70.6% sensitivity. One pneumothorax and two cases of hemoptisies were encountered. COMMENTARY: US guidance is a useful method for the diagnosis of peripheral thoracic masses, yielding a high sensitivity and a very low complication rate. PMID- 8729339 TI - [Cine-MRI of the shoulder. Normal aspects]. AB - PURPOSE: To define the normal patterns of gleno-humeral joint, with kinematic MR imaging in asymptomatic volunteers. MATERIAL AND METHOD: Twenty asymptomatic volunteers (39 shoulders) were studied with a 1.5 Tesla imager. Successive acquisitions Flash (99, 18, 15 degrees) were obtained in the axial plane, from full internal to full external rotation, at the superior, mid and inferior glenoid level. RESULTS: Free edge of the anterior labrum was noted slightly mobile, its base was always immobile. The anterior labrum exhibited modification of shape and increased signal intensity in internal rotation (54%). The posterior labrum remained motionless in 97% and neither shape or signal intensity changes were noted in internal rotation. Inferior labro-capsular ligamentous complex was visualised in all cases. The anterior capsular joint was taut in external rotation and exhibited a slack (51%) or folded (13.5%) pattern during internal rotation. CONCLUSION: Kinematic MR imaging allows a dynamic assessment of the different anatomic components which may be implicated in shoulder instability. It provides information about the labro-capsular ligamentous complex. PMID- 8729340 TI - [Asymptomatic pulmonary embolism and venous thrombosis of the lower limbs. Study with spiral x-ray computed tomography]. AB - AIM: Spiral Computed Tomography (CT) allows diagnosis and follow-up in pulmonary embolism (PE). MATERIAL AND METHOD: We studied prospectively with spiral CT 21 patients to evaluate the frequency and the extent of asymptomatic PE in the patients with lower limbs venous thrombosis and no pulmonary symptomatology. RESULTS: We observed a asymptomatic PE in 8/21 cases (34%). Extensive pulmonary thrombosis could be observed despite normal presentation. There was no association between the localization of PE topography and, the extent of the lower limb venous thrombosis extent. CONCLUSION: This study shows about the capability for CT to evaluate PE severity and clinical underestimation PE occurrence. PMID- 8729341 TI - [Spontaneous remission of renal artery stenosis]. AB - Retrogression of a renal artery stenosis after a six months follow-up is reported in a 43 years old woman who stopped smoking after the first angiography and was treated with antiplatelet, suggesting spontaneous atherosclerosis injury revision. PMID- 8729342 TI - [Spinal osteoid osteoma, neuralgia and MRI]. AB - Computed Tomography (CT) is the most valuable imaging modality for the diagnosis of osteoid osteoma. Magnetic Resonance Imaging (MRI) is less performant than CT. However, in case of neuralgia a disk disease is searched for and MRI is often performed at first. The authors report 2 cases of MRI diagnosis of osteoid osteomas histologically proved. The presence of bone marrow and soft tissues changes consistent with inflammation adjacent to the nidus is the main sign. Inflammatory changes are characterized by low signal on T1-weighted sequence, high signal on T2 and enhancement after gadolinium IV administration. These changes are not specific. But in these 2 cases, an osseous abnormality was detected and the nidus was suspected even if MRI was less conspicuous than CT. In case of children or young adults with radiculalgia and normal disk, inflammatory localized changes in MRI must be suspicious of the diagnosis of osteoid osteoma. PMID- 8729343 TI - [Spontaneous pulmonary hernia and Recklinghausen disease]. AB - The pulmonary hernia is a rare entity defined by protrusion of pulmonary tissue through an abnormal weakness of the thoracic wall. We describe a case of spontaneous pulmonary hernia associated with osseous sternocostal abnormalities in a patient with a Recklinghausen disease. PMID- 8729344 TI - [A rare case of chondroid chordoma of the cervical spine]. AB - Chordoma is a rare tumor in spine. Authors report the case of a 49 years old woman suffering from C5 left neuralgia. Plain films showed an enlargement of C4 C5 left foramina. CT scan permitted to see a lobulated tumor with low density just a few enhancement into septa. MRI showed the tumor with low signal on T1wi, high signal on T2wi and slight enhancement after Gadolinium administration. The extension in the vertebral body is very limited. Differential diagnosis are chondroma or chondrosarcoma and epidermoid cyst. Histology with evidence of a chondroid matrix explain the CT and MR appearance. In this localisation, there is no case reported in the litterature. This is an outstanding case because its extra-osseous localization with a lack of contrast enhancement after injection and its unusual histologic pattern. PMID- 8729345 TI - [Multiple malformation syndrome: situs inversus, multiple spleen and congenital anomaly of the inferior vena cava]. PMID- 8729346 TI - [Histopathology of brain tumors. Histoprognosis and its limitations]. AB - Although the current radiological imaging is really performing, histologic examination remains essential in as much regarding the diagnosis as the pronostical assessment of cerebral tumors pronostic. The difficulties of histological diagnosis are not the same according to the various processes such as cerebral biopsy, extemporaneous examination or routine examination. Grading of gliomatous tumors is one among all the components that might enable pronostical assessment. Predictivity of meningiomas and pituitary adenomas recurrences remain unreliable despite numerous histological techniques that can be used (proliferative markers, bromodeoxyuridine incorporation, nucleolar organizers, flow cytometry). PMID- 8729347 TI - [Clinical presentation and prognosis of brain tumors]. AB - The diagnosis of a cerebral tumor is received as dramatic by the patient or his family on account of the fonctional and symbolic idea of the brain and the perspectives of neurosurgical and psychological disabilities. Nowadays it is simplified because actual techniques of imagery are atraumatic and reliable. So the first signs are not to be undervaluated. Prognostic depends of identifiable factors. They lead the choice of therapy and help the medical doctor for the treatment follow-up and for answering to the patient's life questions. PMID- 8729348 TI - [Imaging of brain tumors in adults by tomodensitometry and magnetic resonance imaging]. AB - Adult brain tumors comprise a remarkably diverse group of histologic varieties; the potential of CT and MRI for diagnosing presence of tumors of the brain and evaluating their extension is well recognized. A precise radiologic diagnosis can be often made as for as extra-axial tumors are concerned. On the contrary this is often more difficult for intraparenchymal tumors. PMID- 8729349 TI - [Surgery of brain tumors]. AB - Treatment of brain tumors represents a very important chapter of neurological surgery. In fact, surgery is of primary importance in the management of benign or potentially anaplastic brain tumors. It not only provides histological diagnosis but radical surgical removal, when feasible, is the goal standard treatment of a very large group of these lesions. Surgery is directed specifically against tumoral processes which are widely different and heterogeneous anatomically and histologically. Some of them are benign and their treatment is essentially relaying on surgery. Others are anaplastic or malignant and surgery has to be associated with other treatment modalities [radiotherapy and (or) chemotherapy] but remains crucial for the histological assessment of the lesion even obtained from stereotactical biopsy specimens. Whenever possible surgical resection must be proposed because it improves the patient's clinical and functional state and allows a better efficacy of complementary treatments when indicated. These factors correlates with the quality and length of the postoperative radiological and clinical remission. PMID- 8729350 TI - [Radiotherapy of brain tumors]. AB - When radiotherapy is employed for central nervous system tumours, several clinical considerations relating to the special normal tissue environment in which they are located deserve recognition. First of all the radiosensitivity of normal brain is nearly equivalent to that of the majority of the primary tumours requiring irradiation. Secondly destroyed normal neural tissue never regenates, but partial recovery is possible following limited injury. Thirdly definition of the target volume is determined mostly by indirect means from a synthesis of neuroradiological findings (C.T. scans, M.R.I.). Finally the rigidity of the intact cranium confers greater clinical significance on mass effects including postradiotherapeutic edema. Most brain tumours respond to external irradiation which may be applied either postoperatively or definitively, e.g. gliomas, lymphomas, medulloblastomas, and metastases. New stereotactic techniques, including radiosurgery, interstitial brachytherapy, and proton beam radiotherapy allow the delivery of larger dose in a limited volume. PMID- 8729351 TI - [Chemotherapy of primary malignant brain tumors in adults]. AB - Chemotherapy of primary malignant brain tumors (PMBT) is palliative, except for germinomas. It is used as adjuvant therapy or alone at recurrence. The chemosensitivity of PMBT differs among tumors of different histological types. The role of chemotherapy in the treatment strategy will be reviewed by tumor type (malignant astrocytic gliomas, anaplastic oligodendrogliomas and mixed gliomas, anaplastic ependymomas, medulloblastomas, germinomas, primary malignant cerebral lymphoma). PMID- 8729352 TI - [Brain metastases and their treatment]. AB - Brain metastases will occur in about 25% of patients with cancer. Lung and breast cancer are the most frequent primary cancers (50%). The treatment of brain metastases is palliative. Solitary brain metastasis should be strongly considered for surgical extirpation followed by whole brain radiotherapy. Patients with poor systemic performance status and/or multiple metastases are candidates for whole brain radiotherapy. The role of chemotherapy and focal radiotherapy in the management of brain metastases is still being evaluated. Appropriate use of therapeutic modalities for brain metastases will ameliorate symptoms in most patients and usually increase survival and enhance the quality of the patient's life. PMID- 8729353 TI - [Biology and gene therapy of glioma]. AB - Although prognosis of malignant gliomas did not change dramatically for the last 15 years, tumorigenesis is much better understood. The study of the genetic alterations occurring in these tumors allowed to recognise the most critical genes involved, which are either overexpressed oncogene, or inactivated tumor suppressor gene. Gene therapy is a natural outcome of such progresses. One way of tumor gene therapy is based on correction of genetic defect either by introducing the missing tumor suppressor gene or by blocking overexpression of an activated oncogene. Alternatively, "destructive" gene therapy is based on a "suicide" gene introduced in tumor cells. These approach results now in phase 1 protocol. PMID- 8729354 TI - [Non-cirrhotic alcoholic liver disease. Pathological anatomy, diagnosis, course, treatment]. PMID- 8729355 TI - [Tetanus. Physiopathology, diagnosis, prevention]. PMID- 8729356 TI - [Anxiolytic drugs. Principles and rules of use]. PMID- 8729357 TI - [Calcium channel inhibitors, nitrate derivative. Principles and rules of use]. PMID- 8729358 TI - [Suicidal ideas or behaviors. Diagnostic orientation]. PMID- 8729359 TI - [Painful knee. Diagnostic orientation]. PMID- 8729360 TI - [Erythema nodosum. Diagnostic orientation]. PMID- 8729361 TI - [Spasm of normal or irregular coronary arteries. Long-term outcome of 277 patients]. AB - The outcome of patients presenting with spasm of normal or subnormal coronary arteries is only known in small series of patients at medium-term. The authors reviewed the outcome of 277 successive patients over an average period of 86 +/- 43 months (12 to 174 months). There were 206 men and 71 women with a mean age of 54 +/- 9 years. Coronary angiography was performed in all cases and did not show any stenosis greater than 50%. Spasm was documented during coronary angiography in 157 cases (57%) by a positive Methergin test after coronary angiography in 113 cases (41%) and by an ECG recording of Prinzmetal angina in 7 cases (2.5%). Nearly all patients (264: 95%) were treated by calcium antagonists. At the end of follow-up, there were: 35 lost to follow-up (12.6%), 20 deaths (7.2%) of which 10 were cardiac (3.6%), 18 myocardial infarctions (6.5%): 11 had repeat coronary angiography which showed one or more new significant (> 70%) coronary lesions in all cases; 109 patients had persistence of chest pain (39%). The severity of symptoms in 52 cases (over one attack of chest pain per month) led to repeat coronary angiography which showed significant coronary disease in 19 cases; 95 patients (34%) were asymptomatic. A multivariate statistical analysis showed hypertension or subnormal appearances on the initial coronary angiography to be significant predictive factors for new coronary events (death, myocardial infarction or angina requiring repeat coronary angiography). The authors conclude that coronary spasm of angiographically normal or subnormal arteries is only well controlled at long-term in 39% of patients not lost to follow-up and is responsible for death or myocardial infarction in 11.6% of cases (nearly 1.5% per year). PMID- 8729362 TI - [Detection of circulating endothelial cells: a new diagnostic test of angina at rest]. AB - The diagnosis of spontaneous angina depends on the recording of per-critical electrocardiographic changes. There is no simple biological test to make its retrospective diagnosis. The attack is usually triggered by instability of an atheromatous plaque which fissures and liberates endothelial cells in the blod stream. The detection of these cells cold therefore be a biological sign of this condition. The technique of detection of circulating endothelial cells by immuno magnetic method was used in 3 groups of patients admitted to hospital within 24 hours: group I comprised 11 patients with acute myocardial infarction, group II comprised 23 patients who had suffered from spontaneous angina with ST segment depression during the attack and significant coronary arterial stenosis, group III comprised 6 patients with chest pain for which coronary angiography is normal and provocative test of spasm is negative. Circulating endothelial cells were detected in all patients of group I (100%), in 18 of the 23 patients of group II (78%) and only in one of group III (18%). These results confer on this biological test for spontaneous angina a specificity and predictive positive value of 83 and 95% and a sensitivity and negative predictive value of 78 and 50%. Therefore the detection of circulating endothelial cells could be used as a simple and reliable test for retrospective diagnosis of spontaneous angina. The mediocre sensitivity and negative predictive value may be explained by a mechanism other than fissuration of atheromatous plaque in some cases of spontaneous angina. PMID- 8729363 TI - [Contribution of doppler echocardiography to the diagnosis of the first attack of acute rheumatic fever]. AB - Since 1944, the Jones criteria for the diagnosis of acute rheumatic fever have been regularly revised to integrate technical and diagnostic innovations. Echographic and Doppler criteria, however, remain unrecognised due to valvular insufficiency in healthy subjects. The aim of this study was to determine the cardiac lesions occurring in acute rheumatic fever and the diagnostic value of Doppler echocardiography. One hundred patients with an average age of 10 years were admitted to hospital because of a first attack of acute rheumatic fever between January 1991 and September 1992. Eighty-six had articular signs, 5 had chorea, but none had cutaneous lesions. Forty-seven murmurs of mitral insufficiency (MI) and eight of aortic insufficiency (AI) were detected; 10 children had signs of cardiac failure. Conduction defects were recorded in 12 cases. Echocardiography showed 7 pericardial effusions; often, the left heart chambers were dilated without alteration of the fractional shortening. The commonest lesions of the mitral valve were thickening of the two leaflets, the reduced mobility of the posterior leaflet, the rigidity of the anterior leaflet and 2 cases of ruptured chordae tendinae. The Doppler mode showed 73 cases of MI, 26 of which were at least moderately severe. These cases of MI were commonly excentric jets behind the posterior leaflet. There were 47 cases of AI, 10 of which were at least moderately severe. If all cases of moderately severe or mild AI and MI are considered as pathological when associated with suggestive morphological valve changes, the number of cases of carditis increased from 50 without the Doppler mode to over 80 with this mode. Doppler echocardiography validated the Jones criteria in 16 children. The authors propose Doppler echocardiography criteria for the validation of carditis. PMID- 8729364 TI - [Carpentier's supra-annular bioprosthesis in aortic position: a mid-term evaluation]. AB - The aim of this study was to assess the medium and long-term results of the Carpentier-Edwards supra-annular (CESA 2650) bioprosthesis. From the end of 1984 to January 1993, 160 patients aged 71.6 +/- 8.6 years underwent aortic valve replacement alone or associated with coronary revascularisation. The postoperative mortality was 11 patients (6.8%): follow-up included 749 patient years with an average period of 60 +/- 29 months, which was complete in 96% of operated patients. The late mortality was 30 patients (4% patients/year): the actuarial 5 year and 10 year survival was 78.6% and 59.1% respectively (hospital mortality included). The linear complications rates were: major thromboembolism: 1.3% patients/year; structural deterioration: 1.5% patients/year; reoperation: 0.75% patients/year; total morbid/fatal complications: 5% patients/year. Despite the limited follow-up, the authors' analysis confirmed the low rate of structural deterioration requiring reoperation, especially in patients over 70 years of age in whom there were no cases of reoperation at 9 years. They conclude that the medium-term results of the CESA 2650 bioprosthesis are comparable to those of first generation porcine bioprostheses: the absence of reoperation related to valve complications in the over 70s during the study period suggests that this is the replacement of choice in this category of patients. PMID- 8729365 TI - [Management of nodal reentrant tachycardia with radiofrequency: predictive criteria of success]. AB - Endocavitary catheter ablation by radiofrequency energy applied on the slow pathway is an effective method of treatment of nodal reentrant tachycardias. The aim of this report was to determine the criteria predictive of success during radiofrequency ablation of the slow pathway guided by the presence of slow potentials. Thirty-five patients (21 women, 14 men, mean age 44 +/- 14 years) with frequent attacks of junctional tachycardia were studied. After confirmation of the diagnosis by electrophysiological investigation, radiofrequency energy was delivered at a site characterised by the presence of slow potentials between the atrial (A) and ventricular (V) potentials. The criteria investigated at each site were: before application: A/V ratio; amplitude of A and V: maximum A/minimum A ratio; amplitude and duration of the A potential; during ablation: radiological stability of the catheter position and occurrence of a junctional rhythm. All 35 patients had successful procedures with no inducible tachycardia at the end of the procedure. The slow pathway was destroyed in 20 cases (57%) with no complication of atrioventricular block. The duration of the A potential was longer in the successful cases (56 +/- 16 vs 48 +/- 14 ms; p = 0.04). The appearance of junctional rhythm and catheter stability during the procedure were predictive of success (79% vs 48%; p = 0.02; 74% vs 43%; p = 0.01). The authors concluded that an ablation site with a long duration A potential and a slow potential is a good target. In addition, ablation should be started when the catheter is radiologically stable and should not be interrupted in the absence of a junctional rhythm. PMID- 8729366 TI - [Atrial electrophysiological study of unexplained ischemic cerebrovascular disorders]. AB - The aim of this study was to search for the presence of atrial vulnerability by programmed atrial stimulation in patients with unexplained ischaemic cerebrovascular strokes and to evaluate the effects of intravenous flecainide acetate on the electrophysiological parameters and on the induction of atrial arrhythmias. Thirty-eight patients (20 men, 18 women) with a mean age of 38.4 +/- 11 years were investigated. Programmed atrial pacing triggered a sustained (> 1 min) atrial arrhythmia with 1 or 2 extrastimuli in 23 of the 38 patients (61%), in these patients, there was a significant shortening of the effective refractory periods (ERP: 193 +/- 23 vs 218 +/- 30 ms; p < 0.02) and of the functional refractory periods (FRP: 228 +/- 25 vs 253 +/- 27 ms; p < 0.01) with lengthening of the A2 auriculogramme (99.7 +/- 22 vs 76.1 +/- 16 ms; p < 0.05). A combined study of the refractory periods and conduction defects provides a means of calculating an index of latent vulnerability which is greatly shortened when an atrial arrhythmia is induced (2 +/- 0.5 cm vs 3 +/- 0.6 cm; p < 0.001). Atrial arrhythmias could not be initiated after intravenous 3 mg/kg of flecainide acetate in 4 subjects (17%) with initially positive atrial stimulation tests. This study confirms the high frequency of atrial vulnerability in patients with unexplained ischaemic cerebrovascular strokes. In their population, the authors observed a low efficacy of flecainide acetate in the prevention of reinduction of atrial arrhythmias. The indication of long-term antiarrhythmic drugs in these patients are questionnable and should be assessed by a prospective long-term multicentre trial. PMID- 8729367 TI - [Indium III monoclonal antimyosin antibody scintigraphy for the detection of chronic myocardial infarction apart from the acute phase]. AB - Fab antimyosin scintigraphy has been shown to be sensitive and specific in detecting acute myocardial necrosis. This study was designed to evaluate the preoperative frequency of Indium-111 (In-111) antimyosin myocardial uptake in patients scheduled for coronary artery bypass surgery. The scintigraphic results were compared with other criteria of myocardial infarction (MI). Sixteen consecutive patients were included. Recent MI (1 to 3 months) were detected in four patients, with an accurate localization in three cases when compared to the classic criteria for MI. Two more patients had old Q wave MI: one did not show any uptake in the territory of MI whereas the second patient with a 21 year old infarct without recent acute coronary events showed an intense uptake consistent with the ECG and angiographic localization. Four other patients with stable angina showed limited uptakes that were unexpected, since there were no acute coronary events in their medical history, and ECG. Their left ventricle angiography were considered as normal. In these four cases, the scintigraphic location corresponded to a territory supplied by an occluded coronary artery (n = 2) or by a coronary artery with a tight stenosis requiring a bypass graft (n = 2). These antimyosin uptakes are probably related to small necroses which did not modify the ECG and did not alter the ventricular segmental wall motion. We conclude: 1) recent MI are detected by In-111 antimyosin scintigraphy; 2) In-111 antimyosin uptake may occur in patients without a diagnosis of recent myocardial infarction and correspond to older MI or limited necroses without detectable changes of the ECG and left ventricle angiography. PMID- 8729368 TI - [Supraventricular tachycardia with wide QRS complexes during Vaughan-Williams class I anti-arrhythmic treatment. Diagnostic and therapeutic implications]. AB - The authors report 8 cases of regular tachycardia with wide QRS complexes during treatment with Vaughan-Williams class 1 antiarrhythmic drugs. These antiarrhythmics, prescribed to prevent atrial fibrillation (3 patients) and atrial flutter (5 patients), were flecainide in 4 cases, propafenone in 2 cases and cibenzoline and hydroquinidine respectively associated with digitoxine and propranolol. These wide complex tachycardias were regular atrial tachycardias with 1/1 conduction to the ventricle. The action of the drug, more pronounced on intra-atrial conduction velocities than on atrioventricular node refractoriness resulted in transformation of flutter at 300 cycles/min with 2/1 conduction and a ventricular rate of 150 cycles/min to atrial flutter at 210 cycles/min with 1/1 ventricular conduction. This acceleration of the ventricular rate was accompanied by widening of the QRS complex. Using the new ventricular tachycardia criteria recently published by Brugada resulted in a diagnostic error in 7 out of the 8 cases. The recording of a wide QRS complex tachycardia in a patient on class 1 antiarrhythmic therapy for an atrial arrhythmia should alert the physician to 1/1 atrial tachycardia despite morphological electrocardiographic criteria of ventricular tachycardia. The 1/1 atrial tachycardia may be poorly tolerated and require emergency treatment. The preventive association of a drug which slows conduction through the atrioventricular node is not always effective. PMID- 8729369 TI - [Infections on permanent endocavitary pacemaker leads: value of transesophageal echocardiography]. AB - The prevalence of infection of permanent pacing material ranges from 0.13 to 19.9% of patients according to published series. The seriousness of this condition requires early diagnosis and treatment. Transesophageal echocardiography visualised vegetations on the intracardiac pacing lead in all of 11 patients studied, whereas transthoracic echocardiography and polynuclear leucocyte scintigraphy only provided positive diagnoses in 4 cases for each investigation. Three types of vegetation were visualised; no cases of tricuspid valve endocarditis were observed. Treatment was based on explantation of all implanted material by endovascular traction in 7 cases and by surgery in the other 4 cases according to the results of transesophageal echocardiography. There were no deaths or recurrence of infection. Transesophageal echocardiography is the investigation of choice for imaging a vegetation on an endocavitary pacing lead. Complete explanation is essential for a complete recovery of this infection. PMID- 8729370 TI - [Thrombolytic therapy of acute myocardial infarction]. AB - Thrombolytic therapy has reduced the mortality of acute myocardial infarction when administered in the first 12 hours after its onset. The author reviews the evidence concerning the more difficult aspects of this treatment: do all patients derive the same benefits? How to administer the treatment as early as possible? What is the value of late thrombolytic therapy? What is the best agent and the optimal mode of administration? This article describes the state of the art at the beginning of 1995 in a rapidly evolving field of therapeutics. PMID- 8729371 TI - [Duration of antivitamin K therapy in venous thromboembolic disease]. AB - The necessity for anticoagulant treatment after pulmonary embolism or deep vein thrombosis has been demonstrated. The modalities of this treatment have been established, especially the value of initial heparin relayed by oral antivitamin K therapy with a target INR value between 2 and 3. The last question remaining in this protocol is that of the duration of anticoagulant treatment. The choice of duration of anticoagulation should take into consideration two potential complications: haemorrhage due to over-anticoagulation and excessive duration of therapy, and recurrent thromboembolism which could result from an inadequate duration of therapy. Several trials have addressed this question and have led to a consensus of opinion: therefore, secondary venous thrombo-embolic disease, occurring under known, special circumstances, the cause of which has been treated, should be given 4 to 6 weeks anticoagulant therapy. In the other cases, so-called idiopathic venous thromboembolism (the proportion of which is on the increase), recent studies are inadequate to reach a consensus. These "idiopathic" forms are characterised by a higher incidence of recurrent thromboembolism of "secondary" cancer and coagulation abnormalities. The search for the optimal duration of anticoagulant therapy in these forms requires prospective trials taking their features into account and should lead to further therapeutic options. The evaluation of longer treatment protocols with less intensive degrees of anticoagulation and of alternatives to oral vitamin K antagonists is justified. PMID- 8729372 TI - [Iatrogenic left coronaro-atrial fistula after mitral valve replacement: apropos of a case]. AB - The authors report a case of an iatrogenic fistula between the left circumflex coronary artery and left atrium. The fistula was a complication of reoperation to replace a mitral valvuloplasty annulus by a mechanical hemi-disc prosthesis (Saint Jude Medical). Diagnosis was made by transoesophageal echocardiography and confirmed by coronary angiography. The patient underwent external ligature under cardio-pulmonary bypass. PMID- 8729373 TI - [Postpericardiotomy syndrome; a rare complication of transcavitary cardiac pacing: apropos of a case]. AB - The authors report a new case of the post-pericardiotomy syndrome occurring after implantation of a DDD pacemaker. This is a rare complication of the insertion of a screw atrial electrode, the iatrogenic potential of which has already been reported. The immediate outcome was favourable with anti-inflammatory therapy. PMID- 8729374 TI - [Syncopal ventricular tachycardia complicating paroxysmal atrial fibrillation in severe myocardial ischemia]. AB - The authors report two cases of spontaneous paroxysmal atrial fibrillation closely followed by syncopal ventricular tachycardia resulting in cardiac arrest requiring defibrillation. Both patients, men aged 62 and 64 years, had a past history of myocardial infarction without arrhythmias; atrial fibrillation occurred during severe myocardial ischaemia; coronary arteriography showed tight stenoses of the left main coronary artery with normal left ventricular function. Ventricular tachycardia (or fibrillation) during spontaneous paroxysmal atrial fibrillation is a rare occurrence. This sequence of events has been described in patients with accessory conduction pathways or in hypertrophic cardiomyopathy. It is an exceptionally rare complication of ischemic heart disease with only a very few previously reported cases. Myocardial ischaemia is probably the cause of the arrhythmia in together with irregularity of the ventricular contractions responsible for long cycle-short cycle sequences which are particularly arrhythmogenic and changes in sympathetic tone. PMID- 8729375 TI - [Deformation of the tricuspid annulus by pericardial adhesions: a rare cause of early tricuspid regurgitation after mitral valve replacement]. AB - The authors report the case of major tricuspid regurgitation occurring early after mitral valve replacement. The mechanism was demonstrated at reoperation: the heart was deformed by a posterior pericardial effusion and cardiodiaphragmatic pericardial adhesions. PMID- 8729376 TI - [Primary vesico-ureteral reflux. Current data and perspectives]. AB - Primary vesicoureteral reflux is a frequent condition threatening the renal parenchyma. This severe disease can lead to renal dysplasia and scarring. When revealed by acute urinary tract infection, parenchymal damage has, in most cases, already occurred so medical or surgical care comes too late. Ideally, the population should be screened at birth for vesicoureteral reflux, but unfortunately, no biological test is yet available. At the present time, it would be reasonable to propose cystography to "at risk" populations, first in newborns with pyelocaliceal dilation detected antenatally then in siblings of patients treated for vesicoureteral reflux. Bladder dysfunction, also often associated with primary vesicoureteral reflux, should be identified early and treated. Finally there are several, often opposing, patterns of clinical expression. Antenatally detected vesicoureteral reflux predominates in the male population. Congenital parenchymal lesions, even in the absence of urinary tract infection, are associated with a small capacity hyperactive bladder. Primary vesicoureteral reflux in the school-age girl is associated with post-infectious renal scarring and with a large bladder incompletely emptied during micturition. Early diagnosis, screening of high-risk populations and prevention of infection together with better management of bladder dysfunction should help reduce the incidence of parenchymal damage due to primary vesicoureteral reflux. PMID- 8729377 TI - [Erythrocyte and plasma antioxidant activity in diabetes mellitus type I]. AB - OBJECTIVES: Some biologic parameters involved in cell defence against oxygen radicals (plasmatic vitamins C and E, erythrocyte glutathione peroxidase, glutathione reductase and superoxide dismutase) were measured in single blood samples from 119 diabetic infants, adolescents and young adults. METHODS: Data were studied in relation to residual insulin secretion determined by C peptide, level of metabolic control appreciated by glycosylated haemoglobin, lipid abnormalities and subclinical complications (retinopathy, neuropathy and nephropathy). RESULTS: There was no change in antioxidant parameters with insulin secretion. Patients with poor glycaemic control and high plasma lipids had higher levels of plasma vitamin E. Patients with nephropathy had lower plasma vitamin C levels and those with neuropathy showed lower erythrocyte glutathione peroxidase activity. Plasma vitamin C concentrations and erythrocyte glutathione reductase activities were negatively correlated with the age of the patients and the duration of the disease. CONCLUSION: Higher transport capacity of vitamin E probably explains the elevated levels of vitamin E observed in patients with high lipid levels and long lasting illness. The lower levels of vitamin C in the presence of nephropathy may be due to an increased renal excretion of this vitamin. The reduction of glutathione peroxidase, glutathione reductase activities and vitamin C levels confirms the existence of an oxidative stress in type I diabetes. PMID- 8729378 TI - [Acid-alcohol-resistant bacilli detected by microscopic analysis of exhaled air from HIV-infected patients: tuberculosis or mycobacteriosis?]. AB - OBJECTIVES: The incidence of atypical mycobacterial infections has increased with the AIDS epidemic. To present, microscopic examinations of airway specimens positive for acid-fast bacteria were highly suggestive of tuberculosis. However, since the AIDS epidemic, certain authors have reported an increase in Mycobacterium avium intracellulare found in respiratory specimens. The aim of this work was to determine what factors might distinguish between these two infections revealed discovery of an acid-fact bacilli. METHODS: Hospital files of all HIV seropositive patients seen between November 1992 and March 1995 and with at least one airway specimen positive for acid-fast bacilli were studied retrospectively. RESULTS: Mycobacterium tuberculosis was isolated in 19 patients, Mycobacterium avium intracellulare in 8 and culosis and M. avium intracellulare. There was no difference for age, sex, geographical origin, transmission mode, antigen positivity, radiologic findings or clinical signs between patients with the different types of mycobacterium. The CD4 count was however significantly lower in patients with an atypical mycobacteriosis (14.5/mm3) than in patients with tuberculosis (91.7 +/- 83.7) (p = 0.004). CONCLUSION: These findings show that in HIV-infected patients with a CD4 count under 100/mm3, the presence of acid-fact bacilli can indicate either M. tuberculosis or M. avium intracellulare. Combined anti-tuberculosis and antimycobacteriosis therapy thus would appear to be justified until the germ can be identified. PMID- 8729379 TI - [Hepatic psittacosis: a case of liver abnormality diagnosed by ultrasonography]. AB - Psittacosis marked by liver and spleen involvement and minimal pericarditis was observed in an 18-year-old patient hospitalized for fever of 1 month duration. At admission, there was no other clinical manifestation and the chest X-ray showed no sign of pulmonary involvement. Liver tests revealed cholestasis. Ultrasonography of the abdomen revealed multiple nodular formations in the liver and spleen, images confirmed on computed tomography. Liver biopsy showed granulomatous with hyperplasia of the Kupffer cells surrounded by healthy tissue. Complement fixation to psittacosis antigen was positive and increased significantly over a 15-day interval. Treatment with tetracycline led to rapid remission of the fever and normalization of the liver tests and hepatic images. Liver involvement in psittacosis is not uncommon, but this is apparently the first case reporting echographic anomalies. The absence of pulmonary involvement has been reported earlier in a few cases. The diagnosis is usually suggested on the basis of epidemiologic criteria and confirmed by complement fixation. PMID- 8729380 TI - [Visceral localizations of cat-scratch disease in an immunocompetent patient]. AB - Locoregional expression of cat scratch disease is well known, but despite advances in microbiology over the last 10 years leading to the description of two new bacteria (Afipia felis, Bartonella henselae) the infective agent responsible for cat scratch syndrome remains unknown. Until the 80s, only one systemic disease was attributed to infection with a germ in the Bartonella genus: trench fever. With the onset of the AIDS epidemic, new clinical syndromes caused by Bartonella bacteria have been described: bacillary angiomatosis, hepatic peliosis, cases of recurrent septicemia, cases of endocarditis, etc. More recently, atypical forms of cat scratch disease including systemic diseases have been reported in immunocompetent subjects. Although quite rare (1% of the cases), such types of expression can raise questions as to diagnosis both in terms of clinical signs and in terms of bacteriological findings. Clinical and experimental data do not provide a clear direction for treatment but would suggest that prolonged use of aminoglycosides is useful. PMID- 8729381 TI - [Cyclic enteral nutrition]. AB - Cyclic enteral nutrition consists in continuous infusion of nutrients with a pump over a 12 to 14 hour period at night. Different reports have demonstrated that cyclic enteral nutrition is well tolerated in malnourished ambulatory patients. The incidence of pneumonia by inhalation in this type of patients is less than 2%. Excepting patients with major amputation of the small intestine and important functional consequences, the increased infusion rate required by cyclic enteral nutrition does not diminish digestive tract absorption making the technique as effective as continuous 24-hour infusion. The main advantages of the cyclic infusion are the preservation of physiological balance between fasting and feeding, improved physical activity during the day with its beneficial effect on protein-energy metabolism, compatibility with oral nutrition during the day in nutrition reeducation programs, and the psychological impact in patients who are free to move about, further improving tolerance. Finally, cyclic enteral nutrition is adapted to enteral nutrition programs conducted in the patient's homes. PMID- 8729382 TI - [Lipoprotein lipase: a multifunctional enzyme in lipoprotein metabolism]. AB - Lipoprotein lipase (LPL) is a rate-limiting enzyme for the hydrolysis of triglycerides. Recently new insights into non-enzymatic functions have emerged. Complete lipoprotein lipase deficiency associated with chylomicronemia is an uncommon (1/10(6) in the general population) autosomal recessive disorder caused by many different lipoprotein lipase gene mutations and is characterized by high fasting plasma triglyceride levels, that can be complicated with acute pancreatitis. To date, about sixty gene mutations have been described throughout the world. Conversely to the homozygous state, the heterozygous state predisposes to a lipid profile that may be atherogenic evenly frequent (approximately 1/500) in the general population. These new clinical and biological insights reinforce the multifunctional role of lipoprotein lipase. PMID- 8729383 TI - [Cytomegalovirus infections during pregnancy]. AB - The incidence of congenital cytomegalovirus infection in France reaches approximately 1,500 infants per year, leading either to death during the first days of life or severe neurosensorial sequellae. This public health problem is of particular importance since primary infections, with the most severe consequences resulting from infections occurring during the first trimester of pregnancy, usually have no clinical manifestations. Viral culture of the amniotic fluid obtained by amniocentesis is required for diagnosis. Ultrasonography can detect malformations. In some of the most severe cases early termination of pregnancy is justified, but the prognosis in less severe cases is not clearly determined. Currently, no in utero treatment has been shown to be effective. PMID- 8729384 TI - [Changes in taste and smell caused by hydroxychloroquine]. PMID- 8729385 TI - [Microscopic periarteritis associated with bone metastases from breast cancer. Fortuitous association or paraneoplastic syndrome?]. PMID- 8729386 TI - [Isolated vomiting revealing hyperthyroidism]. PMID- 8729387 TI - [Staphylococci causing nosocomial infections in pediatric units. Sensitivity to glycopeptides and primary antibiotics]. PMID- 8729388 TI - [Recurrent peripheral facial paralysis. An unusual case]. PMID- 8729389 TI - [Protected distal bronchial lavage: a rapid diagnostic method for fat embolism]. PMID- 8729390 TI - [A new tumor suppressor gene responsible for type 2 neurofibromatosis is inactivated in neurinoma and meningioma]. AB - Neurofibromatosis type 2 (NF2) is an autosomal dominant disease which predisposes to multiple schwannomas, meningiomas and to a lesser extent, to ependymomas. These tumours have been shown to display frequent loss of chromosome 22. Gene defect causing NF2 has been mapped on chromosome 22. Using positional cloning, we and others recently isolated the gene responsible for NF2. Its product displays strong homology with membrane organizing protein suggesting that this protein, called either Schwannomin or Merlin, could act as a bridge between membrane and cytoskeleton. Alterations of the NF2 gene have been identified in NF2 patients and result usually in a truncated, presumably inactive protein. Analysis of tumoral DNA from sporadic schwannomas and meningiomas demonstrates complete loss of function in many cases, providing evidence that the NF2 gene acts as tumor suppressor gene. As a consequence, genetic presymptomatic diagnosis of at risk individuals for NF2 is now possible. PMID- 8729391 TI - [Congenital muscular dystrophy with merosin deficiency: clinical, histopathological, immunocytochemical and genetic analysis]. AB - A selective deficiency of a specific laminin isovariant, merosin made of M, B1 and B2 chains, was found in a series of 17 patients affected with congenital muscular dystrophy (CMD). The merosin deficiency was complete in 15 cases, and almost complete in two cases. An overexpression of the laminin A chain was seen in these biopsies, while B1 and B2 chains were normally expressed. Comparison of the clinical data with a series of 18 "merosin-non deficient" cases showed that the "merosin-deficient" cases were forming a more homogenous group than the "non deficient" one. Hypotonia, contractures, motor development delay were generally more severe in the "merosin-deficient" series of cases. Moreover, white matter alterations were seen in most cases explored by MRI or scan imaging. A genetic linkage with a 6q2 locus, corresponding to the M chain gene localization, was found in a panel of informative families from French and Turkish origin with "merosin deficient" CMD. "Merosin non-deficient" families did not map on this locus. So, the "merosin-deficient" CMD can be considered as a peculiar entity within the group of Congenital Muscular Dystrophies. PMID- 8729392 TI - [Hiramaya disease: disease or syndrome?]. AB - Hirayama's disease, a benign juvenile amyotrophy of "oblique" topography of an upper limb, has been acknowledged in the international literature. 4 cases are reported in this paper. However there present some differences: presence of antigangliosides antibodies in 1 case, cryoglobulinemia in another case developing bilaterally, conduction blocks in a third case. Finally, in the patient illustrating the typical form, the neuroradiological abnormalities considered as characteristic of cervical spine compression by Hirayama are lacking. PMID- 8729393 TI - [Transesophageal echography and systemic ischemic incidences: 235 cases]. AB - From the first of June 90 to the thirty first of January 94, transesophageal echocardiography was performed in 235 consecutive patients (mean age 56+/-16 years), presenting either with cerebral ischemic event (n = 202) or a peripheral arterial embolism (n = 33). All patients had normal echocardiographic and Doppler examinations of the carotid arteries, and transthoracic echocardiography did not show any possible cardiac origin for stroke. Ninety seven patients (41.2%) had documented cardiac disease and/or atrial fibrillation (group 1); 138 patients (58.8%) had no previous cardiovascular history (group 2). Transesophageal echocardiography revealed a possible embolic source in 65.9% of cases (group 1) compared with 29.7% of cases in group 2 (p < 0.001). Intracavitary thrombus and spontaneous contrast in the left atrium were detected only in group 1 (21.6% vs 0%, p < 0.001 and 24.7% vs 0%, p < 0.001 respectively). Patent foramen ovale was more frequent in group 2: 14.5% of cases vs 4.1% of cases, p < 0.01. There was no significant difference between atheromatous aortic plaques and interatrial septam aneurysm incidence in the two groups. Patent foramen ovale and interatrial septal aneurysm were more frequent in group 2: 85% of cases vs 10% of cases in group 1. Transesophageal echocardiography contributes more in patients with a history of cardiac disease. In patients without cardiac disease, patient with foramen ovale and interatrial septal aneurysm were mainly detected: their association represented a risk factor of cerebral ischemic event. Therefore transesophageal echocardiography should be performed in young patients or in case of recurrent event. PMID- 8729394 TI - [Saccadic latencies and unilateral spatial neglect caused by parietal lesions]. AB - Latencies of lateral visually guided saccades were studied in 12 right handed patients who had unilateral ischaemic lesions involving the parietal lobe. The patients were divided into two groups according to the right or left side of the lesions. The severity of unilateral spatial neglect was assessed. In right parietal lesions, the asymmetry of latencies was related to the presence of unilateral spatial neglect and correlated with ist severity. According to the stimulus condition used, we suggest that the asymmetry of latency is explained in terms of an inability to disengage. PMID- 8729395 TI - [Influence of changes in climate and the calendar on the onset of a migraine crisis]. AB - Several factors have been demonstrated to play a role in triggering acute episode of migraine headache. On the basis of observation, others have been suggested but not confirmed. Variations in climate are among the latter. Over a one-year period, we observed a population of 304 patients with migraine residing in a precise climatic zone. The frequency of acute episodes could be analyzed for 148 among them. There were 4,421 acute episodes recorded during the year. The periodicity during the year was compared with climate parameters: temperature (maximal, minimal, average), wind (force and direction), atmospheric pressure, rain, sunshine, relative humidity, and icy, foggy and stormy weather. Moon cycles were also assessed. No correlations were found between climate parameters or variations there of and onset of migraine. Variations in climate would thus appear to have no effect on migraine. Inversely, the results of this study demonstrated a day effect with a higher number of episodes on Saturday and less on Monday and Tuesday. The triggering effect of catamenia on many acute episodes was also confirmed. PMID- 8729396 TI - [Subarachnoid hemorrhage and onset of marginal hemosiderosis caused by a spinal cord cavernoma]. AB - An intradural cavernoma of the spinal cord located at T12 was identified in a 39 year-old man presenting with recurrent episodes of lower back pain on the basis of MRI findings. The patient was hospitalized for severe subarachnoid hemorrhage complicated with hydrocephalia requiring ventriculo-peritoneal draining. Marginal asymptomatic hemosiderosis of the spinal cord above the malformation was observed. The cavernoma which did not involve nervous tissue was removed surgically to avoid the risk of recurrent bleeding and further development of marginal hemosiderosis. PMID- 8729397 TI - [Contribution of position emission tomography in a case of Marchiafava Bignami disease: Morel's laminar sclerosis?]. AB - We report the case of a 26-year old alcoholic woman who abruptly presented with confusion associated with a major hypertonia. She evolved into a chronically vegetative state. Magnetic resonance imaging (MRI) was consistent with the diagnosis of Marchiafava Bignami disease. The corpus callosum first presented an oedematous aspect, then a central atrophy with an axial band. The clinical presentation and the functional imaging strongly suggest an association between the Marchiafava Bignami disease and diffuse cortical lesions, such as the laminar sclerosis of Morel. PMID- 8729398 TI - [Endonasal ethmoidectomy in naso-sinusal polyposis. Results in 110 surgically treated patients]. AB - We assessed retrospectively functional and endoscopic results obtained in 110 patients who underwent endonasal ethmoidectomy (n = 218). The patients were divided into 3 groups according to associated pathology (polyposis alone, asthma without intolerance to aspirin, Widal's disease) and outcome was evaluated after a mean 19.5 month follow-up. Function was improved in 88% of the patients. Endoscopic recurrence was seen in 40% of the patients including 10% with major polyposis. Oedema of the mucose remained in 20% of the patients and the mucosa was normal in 40%. Improvement in patients with Widal's disease was the least favourable among the three groups and was best in those with asthma and no intolerance to aspirin. Post-operative complications occurred in 12.6% of the patients and were severe in 0.9%. These satisfactory results, both in terms of function and the low rate of complications, suggest that the current medicosurgical management should be continued with particular attention to the rate of endoscopic recurrence. PMID- 8729399 TI - [Endonasal treatment of iatrogenic or spontaneous cerebrospinal rhinorrhea of the anterior cranial fossa]. AB - There is a 25% risk of meningitis in case of cerebrospinal fluid rhinorrhea from the anterior cranial fossa. Treatment usually is based on neurosurgery when medical management is unsuccessful. The risk of morbidity and mortality in such operations is important and recurrence is observed in 25% of the cases. The development of endonasal surgery has increased the incidence of iatrogenic breaches but has also allowed the development of new techniques for closing breaches. Certain authors recently published a recent series of patients treated via the endonasal route. We present here six cases of cerebrospinal fluid rhinorrhoea from the anterior cranial fossa treated at the Foch Hospital. Etiology was iatrogenic in 4 cases, trauma in 1 and spontaneous in 1. The operative technique and mid-term results are presented. Cure was achieved in all cases after a mean follow-up of 2 years. One patient with osteopetrosis of the cranial floor who underwent neurosurgical decompression of the optic nerve complained of recurrent rhinorrhoea which could not be confirmed by endoscopy nor by imaging. Early endoscopic treatment of cerebrospinal fluid rhinorrhoea should be the first intention option as it preserves olfactive function, limits operative morbidity and mortality and leaves open the option of neurosurgery in case of failure. PMID- 8729400 TI - [Recurrent or familial facial palsy. Apropos of 6 cases and review of the literature]. AB - About 5 cases of recurrent facial palsy (1 case of familial recurrent facial palsy) and 1 case of non recurrent familial facial palsy, this work analyse the recent knowledges about this subject. Frequency of recurrent palsy is about 10% of the non traumatic facial palsies. A classification is proposed. The prognostic of a recurrence appears similar to the first palsy. Transmission of familial facial palsy seems autosomic dominant or multifactorial. Among the etiology, some like the alternobaric facial paralysis or Melkerson-Rosenthal syndrome could require special preventive or specific therapy. PMID- 8729401 TI - [High resolution T2 weighted MRI sequence and diagnosis of neurinoma of the VIII]. AB - OBJECT: To assess the value of high resolution fast spin echo T2 weighted (HR-FSE T2w) sequence for the diagnosis of acoustic neuroma. Two criteria for normality of the HR-FSE T2w examination are defined. PATIENTS AND METHODS: 114 patients underwent MR imaging at 1.5 T. The protocol included: axial images; 2 overlapped sequences focused on the internal auditory meatus (IAM), HR-FSE T2w acquisition (512 matrix, 3 mm sections with 1.5 mm overlapping); pre and postcontrast T1 weighted sequences focused on IAM. We defined two criteria for a normal HR-FSE T2w image: cerebellospinal fluid visible throughout the IAM, cochleo-vestibular and facial nerves visible in the IAM. RESULTS: 31 examinations were abnormal with HR-FSE T2w sequence, 25 lesions were found on postcontrast T1 weighted sequence (i.e. 6 false-positive results and none false-negative). The sensitivity, specificity and negative predictive value of this HR-FSE T2w sequence were 100%, 97% and 100% respectively. CONCLUSION: the HR-FSE T2w sequence is the first-line method for detecting tumoral lesions of the IAM. If the 2 normality criteria are present on the HR-FSE T2w images, the T1w sequence with gadolinium injection is of no use; the injection of gadolinium remains necessary if only one of these criteria is lacking. PMID- 8729402 TI - [Lymphoepithelial hyperplasia of the parotid gland and HIV infection]. AB - A retrospective review of 10 patients with parotid enlargement is presented. Nine patients were known to be antibody positive for the HIV and one was subsequently tested to be positive. Parotid enlargement was nodular in 9 patients, bilateral in 7, unilateral in 2. One patient had a diffuse enlargement with xerostomia and xerophtalmy. Histological results confirmed begnin lymphoepithelial lesion in all patients. Pathogenesis, diagnostic, pronostic and therapeutic problems are discussed in a literature review. PMID- 8729403 TI - [Peritonsillar abscess. Emergency tonsillectomy?]. AB - The treatment of peritonsillar abcess is still controversial: quincy tonsillectomy, also called tonsillectomy "a chaud" (TC) or interval tonsillectomy, also known as tonsillectomy "a froid" (TF), several weeks after incision and drainage. Of 105 patients presenting with peritonsilar abscess, 53 had tonsillectomy: 27 TC and 26 TF. For each group the following variables were analyzed: age, location of abscess, bacteriology, duration of surgery, amount of per-operative bleeding, post-operative hemorrhage and complications, and length of hospital stay. The location of the abscess was posterior in 33%, and therefore difficult to drain without tonsillectomy. Significant differences in favor of TC were found for: 1) the amount of per-operative bleeding; 2) the occurrence of post-operative hemorrhage; and 3) the duration of hospital stay. We conclude that TC is a safe procedure without increased risks, with less complications, and with a reduced cost. When surgery is indicated in the treatment of peritonsillar abcess, we advocate TC. From our results and the litterature a decision tree for the management of peritonsillar abcess is presented. PMID- 8729404 TI - [Ectopic lingual thyroid. Apropos of 3 cases]. AB - Lingual thyroid is a rare cause of upper airway obstruction, dysphagia or hypothyroidism symptoms. This report describes three cases of lingual thyroid arising in women. One was in pregnancy and developed a lingual goiter and hemorrhages with hypothyroidy. The second case have been diagnosed because of a dysphagia and the third was asymptomatic and have been diagnosed during physical examination for cervical nodes. Diagnosis and possible therapeutic options are discussed regarding these three cases. Surgical therapy is appropriate for patients with clinical signs of upper airway obstruction or when malignant degeneration is suspected. Without of clinical sign, substitutive therapy with thyroid hormone allows the stabilization or the regression of the ectopic thyroid. PMID- 8729405 TI - [2 cases of unilateral nasal obstruction related to rhinolithiasis]. AB - Authors report two cases of 31 and 42 year old women who for many years had a unilateral nasal obstruction: first case due to a large polyp, second one due to a mucosal inflammation but both caused by a rhinolithiasis. The latter is formed over several years around a vegetable seed or a piece of material neglected since childhood. The rhinolithiasis is a calcareous concretion formed in layers visible by X-rays. This type of growth is usually enhanced by weak economic development of certain countries. PMID- 8729406 TI - [Is there a place for arterial chemoembolization of the liver in the treatment for hepatocellular carcinoma?]. PMID- 8729407 TI - [Clinical and virological evaluation of the detection of pre-S1 and pre-S2 antigens in serum from patients with chronic hepatitis B]. AB - OBJECTIVES AND METHODS: We performed a prospective study to determine the clinical and virological significance of pre-S antigen detection in serum samples from patients with chronic hepatitis B virus infection. Four hundred thirty seven consecutive serum samples from 116 patients were tested for the presence of both pre-S1 and pre-S2 antigens by radioimmunoassay using specific monoclonal antibodies. RESULTS: The pre-S1 antigen/HBs antigen ratio, gave an estimation of the number of pre-S1 epitopes expressed on the surface of circulating viral particles, and was positively correlated with the intensity of viral replication intensity (P < 0.05). Moreover, the pre-S1 antigen/HBs antigen ratio was significantly higher in patients suffering from chronic hepatitis associated with viral replication (24% +/- 13); in anti-HBe positive patients, the pre-S1 antigen/HBs antigen ratio was higher in patients replicating a HBe antigen minus variant of the hepatitis B virus and suffering from chronic hepatitis (17% +/- 9) than in asymptomatic HBs antigen carriers (5% +/- 6) (P < 0.05). The pre-S2 antigen/HBs antigen ratio was not correlated with the level of viral replication or with the patient's clinical status. CONCLUSION: This study confirms that pre S1 antigen detection is a reliable marker of hepatitis B virus replication which can be easily performed in chronically infected patients. This assay is especially useful in identifying anti-HBe positive carriers who replicate a minus pre-core mutant and could benefit from antiviral therapy. PMID- 8729408 TI - [Changes in the polymorphism and concentrations of components of the class II complement after orthotopic transplantation of the liver]. AB - OBJECTIVES AND METHODS: The genes of complement factor B, C2 and C4 are located within the major histocompatibility complex class III region on chromosome 6 in man. These components demonstrate a genetic polymorphism which, when determinated, can be used to define complotypes (association of C2, factor B, C4A and C4B allotypes). On the other hand the liver is the main source of the circulating complement component synthesis. The aim of this study was to analyse the kinetics of several complement component (C3, factor B and C4) concentrations in the plasma and to assess changes in the polymorphic pattern of the complotypes after orthotopic liver transplantation. Nephelometry was used for plasma level measurements and factor B, C2 and C4 typings were performed with high voltage electrophoresis or isofocalisation and immunofixation at intervals before, during and after orthotopic liver transplantation in eleven patients. RESULTS: Complotypes changes were observed 24 hours after liver transplantation in all patients. A slight decrease in C3, C4, and factor B plasma levels was observed in the first hours after transplantation. A rapid increase in the levels of these components was observed subsequently, with normalization in less than 15 days. CONCLUSION: These results demonstrate a rapid synthesis of complement components and the changes in complement polymorphic patterns after liver transplantation. PMID- 8729409 TI - [Hormone-mediated calcium responses of rat and human hepatocytes. Study of multicellular systems by videomicroscopy]. AB - OBJECTIVES AND METHODS: Activation of hepatocyte hormonal receptors leads to the mobilization of intracellular Ca2+ which is thought to be an elaborate system for encoding hormonal messages. We studied hormone-induced calcium signals in freshly isolated multicellular systems of normal rat and human hepatocytes. Calcium signals were recorded by videomicroscopy after stimulation with noradrenaline, angiotensin II, and vasopressin. RESULTS: Calcium signals were highly organized in multiplets: the different hepatocytes responded to Ca(2+)-mobilizing hormones in a sequentially ordered manner, with a first, a second (doublets) and a third (triplets) responding cells. This pattern was an intrinsic feature of the multicellular systems, and seemed to be a result of a gradual heterogeneity of the sensitivity of the different cells, to the hormones. The stimulation of the same multiplet with two different agonists and the removal of the hormone during cell responses provides some evidence for the major role of hormonal receptors in this heterogeneity. CONCLUSIONS: Hormone responses in multicellular systems of rat and human hepatocytes are highly elaborate. The density of hormonal receptors could be the major determinant of the sequential pattern of Ca2+ responses. Hormonal receptors may be gradually distributed among the different cells of the multiplets in vitro and along the porto-centrilobular axis in situ. PMID- 8729410 TI - [Factors of response to treatment with interferon-alpha in patients with chronic hepatitis C. Therapeutic perspectives]. PMID- 8729411 TI - [Consensus conference 1994 on treatments of cancer of the rectum: what can we expect from endorectal ultrasonography?]. PMID- 8729412 TI - [Towards an allotype of second generation colon cancer]. AB - A subset of genetic alterations distinguishes two groups of colon cancers. In the first group instability of microsatellite loci due to a defective DNA mismatch repair system is observed. The second group is characterized by recurrent losses of chromosome regions, frequently associated with hyperploidization. We have developed a technique which enables a fine description of allelic losses in this second group of tumours. The typing of 278 loci in 47 hyperploid colon cancers has provided information for an average of 160 loci per tumour. The high frequency of allelic losses on chromosomes 17, 18 and 5 was confirmed thus validating our methodological approach. Several additional chromosome segments were observed lost in over 40% of the cases, suggesting that tumour suppressor genes may map within these regions. Further technical development should contribute to the identification of these genes. PMID- 8729414 TI - [Colonic perforations complicating colonoscopy. Apropos of 15 consecutive cases observed over 16 years]. AB - OBJECTIVES: Our study was undertaken to better assess the circumstances of perforation during colonoscopy. METHODS: All cases observed in our department between 1978 and 1994 were computer recorded. During this period, 5,822 colonoscopies were performed, including 872 with at least one polypectomy. RESULTS: All perforations occurred during colonoscopies performed under anaesthesia (n = 3,373 vs 2,449 without anaesthesia). There were a total of 15 observations of colonic perforations, 1 perforation in 388 colonoscopies. Ten perforations occurred during diagnostic colonoscopies (0.20%), 5 after polypectomy (0.57%). Eight of 10 perforations complicating diagnostic colonoscopies occurred below an impassable stricture. Two patients died from this complication (0.03% of all colonoscopies). CONCLUSIONS: Our study suggests that endoscopic exploration of colonic stricture, in addition to anaesthesia, increases the risk of colonic perforation. PMID- 8729413 TI - [Variety of pancreatic lesions observed in von Hippel-Lindau disease. Apropos of 8 cases]. AB - Von Hippel-Lindau's disease is a rare genetic disease, with an autosomal dominant mode of inheritance, characterised by the development of several tumours, such as haemangioblastoma of the central nervous system and retina, renal cysts or carcinoma and pheochromocytoma. Several pancreatic lesions, mainly represented by multiple cysts, are also encountered. We report here 8 cases of pancreatic involvement in patients affected with von Hippel-Lindau's disease. It consisted of multiple cysts (3 cases), serous cystadenoma (2 cases), endocrine tumour (1 case), haemangioblastoma (1 case) and ductal adenocarcinoma (1 case). Diagnosis of a rare lesion of the pancreas (multiple cysts, serous cystadenoma or vascularized tumour) in a young patient may lead to search the other lesions of von Hippel-Lindau's disease and to undergo a familial inquiry in order to propose a multidisciplinary approach for patients affected by the disease. PMID- 8729415 TI - [Medico-economic aspects of the treatment of duodenal ulcer: from antisecretory treatment to eradication of Helicobacter pylori]. AB - OBJECTIVES: Radical changes are currently taking place in the treatment of duodenal ulcer, where strategies of eradication of Helicobacter pylori are replacing traditional antisecretory treatment. The aim of this study is to present the economic aspects of the different therapeutic approaches. METHODS: A critical analysis based on a selected number of clinical trials and medical economic analyses published in the international literature. RESULTS: A comparative analysis of traditional ranitidine/omeprazole treatments shows that ranitidine is more cost-effective in France at short term. With regard to traditional strategies, there are no French studies comparing H2-antagonists to proton pump inhibitors. Comparative studies seem in favour of eradication of Helicobacter pylori over traditional therapies, both in terms of cost and effectiveness. CONCLUSION: Arguments comparing the effectiveness of proton pump inhibitors versus H2-antagonists have become obsolete. In current therapeutic approaches of duodenal ulcer, the level of inhibition of the acid secretion no longer plays an important role. PMID- 8729416 TI - [Treatment of carcinoid tumors of the digestive tract]. PMID- 8729417 TI - [Treatment of a hemorrhagic duodenal varice by endoscopic sclerotherapy]. AB - We report the case of a duodenal varix rupture in a 37-year-old man revealing an alcoholic cirrhosis. Endoscopic diagnosis of this duodenal varix was difficult because of its atypical and changing appearance. Endoscopic sclerotherapy was completely successful and there was no recurrent bleeding. Although duodenal varix is rare, this case and the literature emphasize the importance of considering this diagnosis in all patients with duodenal tumoral lesions and suspected portal hypertension. In this context, duodenal biopsy can be dangerous and should be avoided. In case of duodenal varix rupture, endoscopic sclerotherapy appears to be a safe and efficient first-choice therapy. PMID- 8729418 TI - [Endoscopic diagnosis of a biliodigestive fistula of tuberculous origin revealing acquired immunodeficiency syndrome]. AB - We report the case of a 32-year-old Malian man with abdominal tuberculosis revealing acquired immunodeficiency syndrome. A gastroscopy was made for epigastric pain and showed caseum in a digestive fistula with acid fast bacilli. Mycobacterium tuberculosis infection was confirmed by sputum culture. An early antituberculous therapy was prescribed. Outcome was good with rapid fistula closing and slower mass diminution of the abdominal lymph nodes. This case report confirms nodal tuberculosis as a possible cause of digestive fistulae. Rapid endoscopic diagnosis of this tuberculous fistula led to diagnosis of acquired immunodeficiency syndrome and early adapted medical treatment without invasive diagnostic methods. PMID- 8729419 TI - [Still disease in adults revealed by a digestive manifestation]. AB - Adult Still's disease is characterized by typical spiking fever, oligopolyarthritis, neutrophilic leukocytosis and involvement of various organs. We report a case which illustrated typical digestive features of Still's disease as dysphagia, peritonitis and manifests the hitherto unreported complication of gastric ulcerations. Treatment with prednisone was started in order to control arthritis, resulting in improvement of both gastric ulcerations (partially resistant to omeprazole treatment) and arthralgia. After seven years of follow up, the patient remains clinically and biochemically stable with steroid and methotrexate treatment. PMID- 8729420 TI - [Cholangiocarcinoma: a rare cause of chylous ascites]. PMID- 8729421 TI - [Attempted suicide during treatment of chronic viral hepatitis C with interferon. Apropos of 2 cases]. PMID- 8729422 TI - [Acute hepatitis associated with microvesicular steatosis induced by Atrium]. PMID- 8729423 TI - [Hepatomegaly revealing Noonan syndrome without cardiac malformation]. PMID- 8729424 TI - [Favourable course of a pseudotumoral form of hepatic and splenic sarcoidosis under treatment with hydroxychloroquine and colchicine]. PMID- 8729425 TI - [Tuberculosis: an unusual tumor of the esophagus]. PMID- 8729426 TI - [Characteristics of Helicobacter pylori infection in the Noumea Hospital Center]. PMID- 8729427 TI - [Gastric rupture during colonoscopy following oxygen therapy by nasal intubation]. PMID- 8729428 TI - [Partial regression of a desmoid tumor after prolonged treatment with lanreotide]. PMID- 8729429 TI - [Colonic metastases from carcinoma of the piriform sinus]. PMID- 8729431 TI - The isolated perfused "working" rat heart: a new method. AB - This paper presents a "working" isolated perfused rat heart. A special double cannula was designed consisting of an outer cannula that is inserted in the aorta and an inner cannula that is advanced into the left ventricle. The perfusion fluid flows through the inner cannula into the left ventricle, and is ejected from there into the aorta. If the outer cannula system is closed, the fluid perfuses the coronary vessels and drips off outside the heart. When the outer cannula is open and certain pressure resistance is applied, a fraction of the ejected fluid perfuses coronary vessels and the rest is expelled. Because the inner cannula can easily be retracted into the outer cannula, which is placed in the aorta, this preparation provides an opportunity to use the same heart as a "working" or "non-working" model for investigating functions of the heart. The effects of epinephrine and yohimbine were tested on this model. This preparation is simple, durable, and reliable. PMID- 8729432 TI - Estimation of the binding affinity constants of soluble ligand-receptor complexes by a rapid filtration technique: [3H]-flunitrazepam-bovine serum albumin as an example. AB - A method for determining the equilibrium dissociation constant (KA) of a soluble ligand (L) from a soluble receptor (A) in the presence of another solid phase receptor (R) for the same ligand was developed. The total and nonspecific binding of L to R was measured in the presence and in the absence of A. The separation of bound and free L was done by a rapid filtration technique so that only the complex RL, but not AL, was recovered. An apparent dissociation constant (KR,app) was calculated from the saturation curve obtained in the presence of A. The magnitude of KA could be determined from this KR,app and the value of the equilibrium dissociation constant of the complex R-L (KR) calculated from the saturation curve in the absence of A. The equality of the Bmax values obtained in the presence and in the absence of A assured the accuracy in the determination of KA so that the fulfillment of this condition could be used as an internal control. For the correct definition of nonspecific binding, the displacement agent (L1) should be used at concentrations within the range 10(2).KR < L1 < 10. K4. This fact constraints the applicability of the method to systems where KA/KR > 10(3). The highest sensitivity of the method can be attained when 0.33 < [At]/KA < 3. The equilibrium binding constant of [3H]-flunitrazepam to non delipidized bovine serum albumin determined by the present approach (31 +/- 7 mumol/L) did not differ significantly from the literature. PMID- 8729433 TI - Insulin sensitivity tested with a modified euglycemic technique in cats and rats. AB - A new insulin sensitivity test (IST) is described using a modified euglycemic clamp in cats and rats. The IST uses the amount of glucose required to be infused to maintain euglycemia over a 30-min period in rats and 60 min in cats following a bolus administration of insulin as the index of insulin sensitivity. Glucose levels are determined at short time intervals (2-5 min), and variable glucose infusion is used to hold glucose levels within a few percentage points of the basal pre-test glucose level. A new blood sampling procedure is described that allows each IST to be carried out using a total of only 0.5 mL of blood. The IST is sensitive and allows clear insulin dose effects to be demonstrated with 100 mU/kg requiring 355.0 +/- 14.3 mg/kg over 30 min and 50 mU/kg requiring 198.7 +/- 11.1 mg/kg. Five consecutive tests were reproducibly carried out (%CV = 3.0 +/- 0.5) over a 12-hr period in the cat with insulin, glucagon, and glucose levels remaining stable prior to each IST. Glucagon and norepinephrine plasma concentrations do not change significantly during the IST. The IST is sufficiently sensitive to allow demonstration of dose-response relationships for atropine-induced insulin resistance. The IST is thus sensitive, reproducible, and able to demonstrate acute insulin resistance in anesthetized cats and rats. The test is demonstrated in fed (rats) and fasted (cats) state. PMID- 8729434 TI - A novel renal hypertensive guinea pig model for comparing different inhibitors of the renin-angiotensin system. AB - The present study describes a novel renal hypertensive guinea pig model for comparing different inhibitors of the renin-angiotensin system (RAS). Renal hypertension was induced by a two-step procedure consisting of ligation of the left caudal renal artery and right nephrectomy. Sham-operated animals were used as controls. Arterial blood pressure and heart rate were monitored in conscious animals. Left caudal renal artery ligation and subsequent right nephrectomy led to a significant increase (32% over sham-operated controls, p < .05) in mean arterial blood pressure (MABP), 3 to 4 weeks following surgery. Renal hypertensive animals had increased urine production (from 63 +/- 8 mL/kg per day to 143 +/- 29 mL/kg per day, p < .05) and an increased incidence of proteinuria (11/13 animals had urine protein levels higher than 20 mg/kg per day). Five of the 13 renal hypertensive animals also had hematuria. On autopsy, an 83% increase in the left kidney/body weight ratio and a 37% increase in the heart/body weight ratio were observed in the renal hypertensive animals, compared to the sham operated controls. Changes in blood pressure and heart rate were assessed before and after an intravenous bolus injection of the drug to be tested. Captopril reduced MABP in both sham-operated and renal hypertensive animals with equal efficacy (up to a maximum of 42%). In contrast, BILA 2157 BS, one of our human renin inhibitors, produced a similar maximum MABP decrease but only in renal hypertensive animals. This selective antihypertensive effect was also observed with enalkiren, another renin inhibitor. These results indicate that the renal hypertensive guinea pig is an useful model for comparing and contrasting different RAS inhibitors. PMID- 8729435 TI - Automatic leukocyte differentiation in bronchoalveolar lavage fluids of guinea pigs and brown-Norway rats. AB - Asthma is considered to be a chronic inflammatory response of the airways characterized by a leukocyte infiltration into the lungs. Whereas lymphocytes and macrophages are involved in the initiation and propagation of inflammation, both neutrophils and in particular eosinophils are considered to play major effector roles. Therefore, allergic animal models in various species have been established to assess leukocyte infiltration by bronchoalveolar lavage (BAL) of antigen sensitized and antigen-challenged animals as an inflammatory parameter in asthma pharmacology. Differential leukocyte counts in BAL fluids are routinely assessed by visual microscopic analysis of stained slides after cytocentrifugation. This procedure is very time-consuming, and the underlying standard morphological criteria may vary between different observers. In the present paper, we propose an alternative automatic method for leukocyte differentiation in BAL fluids from ovalbumin-treated guinea pigs and Brown-Norway rats using Cobas Helios 5Diff from Hoffmann-La Roche. BAL samples are directly applied to the analyzer and are automatically mixed with "Eosinofix," which stabilizes leukocyte membranes and specifically stains eosinophils. By a combination of electric (resistance) and optical (light scatter) analysis, the lymphocytes, monocytes/macrophages, neutrophils, and eosinophils are discriminated and the total leukocyte numbers are obtained. For both animal species we found high correlations for all leukocyte populations by comparing the results obtained with Cobas Helios 5Diff and conventional microscopic analysis. The major advantage of the automatic method is the much lower (about one-third) time requirement. PMID- 8729436 TI - A highly reproducible model of arterial thrombosis in rats. AB - The objective of this investigation was to develop a reproducible and reliable method of arterial thrombosis in a small laboratory animal. Rats were anesthetized with urethane, and a common carotid artery was exposed. A completely occlusive thrombus was produced by applying an electric current to the arterial wall (2 mA for 5 min) while simultaneously constricting the artery with a hemostatic clamp placed immediately downstream from the electrodes. A complete and persistent cessation of blood flow was obtained in all the control rats starting 10-15 min after the thrombogenic lesion. Histological examination revealed a picture of mixed white and red thrombus, stratified and rich in platelets aggregates and fibrin, with piles of red cells trapped in the fibrin network. On the other hand, stasis alone (clamping) was ineffective at all, whereas electric current application alone caused non-occlusive thrombosis only in 30% of animals. An antithrombotic dose of heparin (3 mg/kg i.v., 30 min before thrombus induction) prevented the formation of a persistent thrombus, blood flow being progressively restored (up to 59% of basal value within 45 min). Similarly, a thrombolytic dose of urokinase-type plasminogen activator (2 mg/kg, for 120 min, starting 15 min after thrombus induction) caused a rapid and progressive resumption of blood flow (up to 80% of basal value). This method gives highly consistent and reproducible results and may be suitable for the study or the screening of antithrombotic as well as thrombolytic agents. PMID- 8729437 TI - A new method to measure cardiac inositol levels in intact animals. AB - Inositol levels have been studied in cellular cultures and recently by perfusion of isolated hearts. The study was aimed to assess inositol turnover in rabbit hearts from intact animals. Thirty rabbits were injected i.v. three times (every 12 hr) with 25 microCi/kg of myo-3H-inositol. The rabbits 12 hr after the last injection were killed and the hearts perfused according to Langerdorff technique. Systolic and diastolic ventricular pressures (SVP, DVP), dp/dt, and coronary flow (CFl) were measured. The hearts (n = 14) were perfused under aerobic conditions and 16 hearts under ischemic conditions for 30 min. In addition, 5 hearts were perfused under aerobic conditions for 10 min, and 6 hearts were perfused under ischemic conditions for 10 min. Samples of myocardial tissue were taken from both groups at the end of 10-min and 30-min period of perfusion, and cAMP and inositol phosphates were assayed. The hearts subjected to ischaemia showed changes of cAMP and 3H-inositol. The cAMP was higher in the ischaemic (10 min and 30 min) than the control hearts, 0.22 +/- 0.09 and 0.21 +/- 0.08 versus 0.41 +/- 0.12 and 0.49 +/- 0.11 pmol 10(6) cells, respectively (p < .05, p < .001. The inositol trisphosphate was higher in control than ischemic hearts (10 min, 30 min), 0.42 +/- 0.02 and 0.39 +/- 0.01 versus 0.31 +/- 0.01 and 0.23 +/- 0.02 (percent of radioactivity) respectively, p < .001. Our data suggest that 3H-inositol may be studied by i.v. administration to intact animals. The ischemia was performed to verify the validity of this new technique. PMID- 8729438 TI - A sensitive radioimmunoassay optimized for reproducible measurement of rat plasma insulin. AB - There has been a notable lack of consistency in plasma insulin values reported in the literature. In an attempt to find a more reproducible and sensitive insulin radioimmunoassay (RIA) method for the measurement of plasma insulin, the charcoal adsorption method for separation of free from antibody-bound antigen was investigated. The applicability of this method towards the measurement of plasma insulin, however, has been known to be severely limited because of interference by plasma proteins with charcoal binding. Indeed, the assay resulted in negative fasting insulin values and unacceptable interassay variability, which was not improved by adding charcoal-extracted plasma (CEP) to the standard curve. In this paper, we demonstrate a process by which relatively small volumes (25 microL) of plasma from control, diabetic, and fasted rats can be assayed reproducibly with charcoal in the final separation step. It was observed that the assay of insulin free plasma, in the form of CEP, could represent a distinct "zero insulin" level specific for plasma samples, in place of the zero insulin standard (insulin-free standard buffer) routinely used. Normalizing plasma samples against CEP resulted in lower intra-assay and interassay variability (coefficient of variation < 10%) and enhanced sensitivity to 7 microU/mL. Recovery was tested by adding 25 microL of known insulin standards to plasma samples in which insulin levels had been determined previously. Whereas the recovery of total insulin in the presence of plasma was low (38%-89%), recovery of total insulin from two standards was > 95%. Following correction, recovery of total insulin in 25-microL plasma was improved to > 90%. Hence, this approach provides a simplified way of correcting for plasma effects and greatly enhances the feasibility of using the charcoal-separation technique for the measurement of plasma insulin. PMID- 8729439 TI - Signal transduction in plant immunity. AB - Significant recent advances in the understanding of plant defense mechanisms include the isolation and characterization of resistance genes against bacterial, fungal and viral pathogens, the identification of genes involved in cell death, and the demonstration of the involvement of reactive oxygen species and salicylic acid in the signal-transduction pathways for expression of induced resistance. PMID- 8729440 TI - Innate immunity in higher insects. AB - The hallmark of the innate immune response of higher insects is the rapid and transient synthesis of a battery of broad spectrum antimicrobial peptides by the fat body. The control of the genes encoding these peptides involves cis regulatory promoter elements homologous to sequences functional in mammalian acute-phase genes. Study of immune-deficient mutants of Drosophila has indicated that distinct pathways control the antibacterial and antifungal responses in this species. Novel receptors potentially involved in the initiation of the immune response have been recently characterized. PMID- 8729441 TI - Immunity to eukaryotic parasites in vector insects. AB - Mosquitoes and blackflies have been the focus of recent efforts to elucidate factors influencing the susceptibility of vector insects to metazoan and protozoan parasites of medical significance. Vector species exhibit variation in cellular and humoral immune responses, as highlighted by studies of melanotic encapsulation and components of the phenoloxidase system. Significant progress has been made in the development of genetic maps based upon molecular markers, leading to the genetic analysis of loci influencing susceptibility. The identification of specific inducible antibacterial peptides, and the cloning of genes encoding immune effector proteins as well as potential regulatory factors, open the path to fruitful studies of vector insect innate immunity and its relationship to insect-parasite interactions. PMID- 8729442 TI - Scavenger receptors in innate immunity. AB - Scavenger receptors are cell-surface proteins expressed by mammalian monocytes and macrophages and by invertebrate hemocytes, among other cell types. They exhibit distinctive ligand-binding properties, recognizing a wide range of ligands that include microbial surface constituents and intact microbes. The ligand-binding properties and expression patterns of these receptors suggest that they may function in one or more host-defense-related processes. Significant advances in scavenger receptor biology have recently been reported, including the identification of several new scavenger receptor genes. PMID- 8729443 TI - The collectins in innate immunity. AB - The collectins are proteins with collagen tails and globular lectin domains that appear to play an important role in mammalian first line host defense. Recent insights have clarified the structural basis of ligand recognition, the interactions of collectins with complement cascades, and the association with disease susceptibility. PMID- 8729444 TI - Mechanisms of phagocytosis. AB - Recent advances in research on phagocytosis include a better appreciation of the cross-talk between phagocytic receptors, the definition of multiple signaling domains within these receptors, and a deeper understanding of the downstream effector pathways leading to actin polymerization and particle internalization. Phagosome maturation in macrophages proceeds via a series of membrane fusion and fission events, which modify the phagosome in small increments, and appears to be regulated, in part, by GTP-binding proteins and perhaps by protein kinase C. The isolation of dysphagic mutants of Dictyostelium discoideum presages the identification of new genes required for phagocytosis. PMID- 8729445 TI - The role of hemolymph coagulation in innate immunity. AB - Invertebrate animals, which lack adaptive immune systems, have developed defense systems that respond to common antigens on the surface of potential pathogens. Hemolymph coagulation is one such defense system in innate immunity. The discovery of lipopolysaccharide-sensitive and (1-->3)-beta-D-glucan-sensitive serine protease zymogens in horseshoe crab (limulus) hemocytes, both of which trigger the coagulation cascade, has exemplified how the animals detect and respond to foreign materials. PMID- 8729446 TI - Developing and shedding inhibitions: how MHC class II molecules reach maturity. AB - Over the past year, several important advances have been made in understanding the mechanisms by which class II MHC glycoproteins acquire endosomal peptides inside antigen-presenting cells. Recent progress in the study of class II antigen presentation includes the identification of ligands from which invariant chain protects class II molecules in pre-endosomal compartments, an improved understanding of how invariant chain inhibits antigenic peptide binding, and the appreciation that HLA-DM (a factor important for antigen presentation in vivo) can act as a catalyst for peptide exchange. PMID- 8729447 TI - Processing and delivery of peptides presented by MHC class I molecules. AB - Effective MHC class I peptide loading requires the proteolytic degradation of cytosolic proteins and the TAP-mediated translocation of peptides across the membrane of the endoplasmic reticulum. The proteasome is emerging as the main cytosolic protease generating class I binding peptides. The recent elucidation of the proteasome crystal structure, together with the use of functional inhibitors, has enhanced our understanding of proteasome function. Genetic analysis of a novel mutant cell line emphasizes the importance of the TAP-class I interaction in the assembly of mature class I heterotrimers, and suggests that additional MHC encoded components are required. PMID- 8729448 TI - T-cell epitope determination. AB - Synthetic approaches to T-cell epitope determination have recently been developed that complement the search for natural T-cell epitopes and the investigation of the preferences of the different MHC alleles for particular motifs in cognate peptide sequences. The combination of these different strategies opens new possibilities for basic, as well as for applied, immunology. The outlines of the strategies for determination of natural T-cell epitopes are well established. These strategies have contributed substantially to our understanding of the nature of T-cell epitopes and of many diseases. Positional scanning approaches with random synthetic peptide libraries allow comprehensive surveys of the sequence requirements for peptide selection by MHC molecules and for induction of T-cell responses. Synthetic T-cell epitopes can be determined independently of the knowledge of the natural T-cell antigen. This opens new perspectives for the development of synthetic vaccines, TCR antagonists and MHC blockers. PMID- 8729449 TI - Human minor histocompatibility antigens. AB - Disparities in minor histocompatibility antigens between HLA-matched organ and bone marrow donors and recipients create a potential risk for graft failure and graft-versus-host disease. These conditions necessitate lifelong pharmacological immunosuppression of organ and bone marrow transplant recipients. Recent technical advances have resulted in the identification of the chemical nature of the first human minor histocompatibility antigens. A new era of research has begun to provide insights into the genetics of minor antigens and their putative role in transplantation. PMID- 8729450 TI - Antigen presentation by CD1 and MHC-encoded class I-like molecules. AB - Three known lineages of antigen-presenting molecules restrict T-cell responses to microbial antigens: MHC class I and MHC encoded class I like molecules present peptides derived from the proteolysis of intracellular pathogens, MHC class ii molecules present peptides derived from the proteolysis of extracellular pathogens and CD1 molecules present unique microbial lipids and glycolipids. Recent studies have indicated that CD1 molecules mediate a novel system of antigen presentation and that MHC-encoded class I-like molecules can present unique subsets of intracellularly derived peptides. PMID- 8729451 TI - Affinity maturation and class switching. AB - Affinity maturation and class switching of antibodies are temporally, but not mechanistically, related processes. The basis of affinity maturation is the selection, in the germinal centers, of antibodies that bind the antigen better. Early in an immune response, the selection is from the primary repertoire; later, it is from mutants generated by hypermutation at the immunoglobulin loci. Recently, the door has been opened for the study of the molecular mechanism of hypermutation, which is expected to make a major contribution to general biology. Class switching has been studied in the past for its obvious clinical importance, but also at the basic level of DNA recombination. Progress in understanding class switching has been trailing the progress made in V(D)J recombination, but new in vitro systems and gene-targeted mice are closing the gap. PMID- 8729452 TI - Biophysical studies of T-cell receptors and their ligands. AB - Recently developed methodologies for the production of the soluble extracellular domains of alpha beta TCRs have allowed several biophysical characterizations. The thermodynamic and kinetic parameters associated with specific ligand interactions between the TCR and MHC-peptide complexes, as well as superantigens, are now being established. Crystallographic studies of isolated TCR fragments have yielded the structures of a V alpha domain and the two extracellular domains of a beta-chain. These investigations are beginning to allow a new visualization of antigen recognition and T-cell activation processes. PMID- 8729454 TI - Innate immunity. PMID- 8729453 TI - Natural killer cell receptors specific for MHC class I molecules. AB - Human natural killer cell receptors for MHC class I molecules have been recently identified by cDNA cloning. They are structurally distinct from murine natural killer cell receptors and can be either inhibitory or stimulatory. Despite structural differences, murine and human receptors are diverse and specific for distinct class I ligands. Their characterization provides insight into natural killer cell functions. PMID- 8729455 TI - Antigen recognition. PMID- 8729456 TI - Expression of poplar phenylalanine ammonia-lyase in insect cell cultures. AB - A cDNA encoding one of the phenylalanine ammonia-lyase genes from Populus trichocarpa x deltoides was inserted into a baculovirus expression vector and the PAL protein was successfully expressed in insect cell cultures. High levels of active holoenzyme were obtained that could be purified in a single chromatographic step. Site-directed mutagenesis and expression of the mutant enzyme confirmed that conversion of the putative active site serine202 residue to alanine is sufficient to destroy the catalytic activity of PAL. PMID- 8729457 TI - Primary structure of trypsin inhibitors from Sicyos australis. AB - Three trypsin inhibitors from Sicyos australis, have been isolated, purified and sequenced. Following protein extraction with ammonium sulphate, the mixture of inhibitors was separated from other proteins by trypsin-affinity chromatography. Subsequent purification of the individual inhibitors was accomplished by reversed phase HPLC. The primary structures of each inhibitor were elucidated by a combination of protein sequencing and electrospray ionization mass spectrometry (ESI-MS) and tandem mass spectrometry (MS-MS) on both the untreated and the reduced and S-carboxymethylated inhibitors. All three inhibitors show extensive sequence similarity with inhibitors from cultivated Cucurbitaceae species, although there are a number of novel residues present. One of the inhibitors has a blocked N-terminus (pyroglutamic acid) and the use of MS-MS was crucial to the elucidation of its primary structure. ESI-MS was further used to characterize the non-covalent complex between one of the inhibitors and trypsin. PMID- 8729458 TI - Insecticidal properties of mushroom and toadstool carpophores. AB - In order to find compounds with insecticidal or antifeedant properties from mushrooms and toadstools, a wide screening was undertaken using the non mycophagous Drosophila melanogaster as a model insect. Powdered fruit bodies of edible and poisonous mushrooms were incorporated with the Drosophila's rearing medium, and their development was observed. Among the 175 different species of fungi tested, 79 were found to inhibit insect development, hence making the isolation of new compounds look hopeful. PMID- 8729459 TI - O-acetylated xyloglucan in extracellular polysaccharides from cell-suspension cultures of Mentha. AB - Extracellular polysaccharide produced by suspension-cultured Mentha cells consisted of 50% neutral sugars, 32% uronic acid and 10% of protein. The ammonium oxalate-soluble fraction of this polysaccharide contained 30% hemicellulose and 70% pectic substances. The purified hemicellulose contained xylose, glucose, arabinose, galactose, mannose and fucose residues in a molar ratio of 41.6:31.3:13.1:11.1:1.3:1.6. It was identified as a xyloglucan from its neutral sugar composition and by methylation analysis and cellulase treatment: the principal neutral sugar was arabinose. The presence of O-acetyl residues was confirmed with 1H NMR, 13C NMR and GC-mass spectrometry. The total acetyl content in the polysaccharide was 4%. The point of attachment of the O-acetyl residue was shown to be at position 6 of the galactosyl residue. PMID- 8729460 TI - Cytotoxic cycloartanes from Aglaia argentea. AB - Aglaia argentea leaves yielded three new cycloartanes: argenteanones A and B, and argenteanol. The first two possess cytotoxic activity against KB cells (IC50 7.5 micrograms ml-1 and 6.5 micrograms ml-1 respectively). Structural elucidation was done by two-dimensional NMR spectroscopy. PMID- 8729461 TI - Further polyoxypregnanes from Marsdenia tenacissima. AB - Two new polyoxypregnanes, designated marstenacigenins A and B, along with a known compound, dresgenin, were isolated from the mild acid hydrolysate of the ethanol extract of the stems of Marsdenia tenacissima. Their structures were deduced by a combination of 1D and 2D NMR spectroscopic techniques as 12 beta-cinnamoyl dihydrosarcostin and 12 beta,20-dibenzoyldihydrosarcostin, respectively. PMID- 8729462 TI - Triterpenoid saponins from Bellium bellidioides. Structures of the minor deacylsaponins. AB - A new deacylsaponin of polygalacic acid, desacylbellidioside B4, was obtained from the whole plants of Bellium bellidioides L. The structure has been elucidated by a general strategy involving mass spectrometry (ESI-MS, including tandem MS, and GC-MS) and high-field one- and two-dimensional NMR spectroscopy (1H and 13C NMR, COSY-45, HMQC, HMBC) as 3-O-alpha-L-rhamnopyranosyl-2 beta, 3 beta, 16 alpha, 23-tetrahydroxyolean-12-en-28-oic acid 28-O-alpha-L rhamnopyranosyl(1-->3)-beta-D-xylopyranosyl(1-->4)-alpha-L- rhamnopyranosyl(1- >2)-[alpha-L-arabinofuranosyl(1-->3)-beta-D-fucopyran oside. Moreover, bellissaponin BS2 and besysaponin C12 have also been isolated, demonstrating the close chemical relationship of B. bellidioides to the Bellis genus. PMID- 8729463 TI - Steroidal saponins from the underground parts of Chlorophytum comosum and their inhibitory activity on tumour promoter-induced phospholipids metabolism of HeLa cells. AB - Three new spirostanol pentaglycosides embracing beta-D-apiofuranose were isolated from the fresh underground parts of Chlorophytum comosum together with four known saponins. The structures of new compounds were determined by spectroscopic data, including two-dimensional NMR, and partial acid-catalysed hydrolysis to be (25R) 5 alpha-spirostane-2 alpha,3 beta-diol 3-O-[O-beta-D-glucopyranosyl- (1-->2)-O-[O beta-D-apiofuranosyl-(1-->4)-beta-D-glucopyranosyl-(1 -->3)]-O- beta-D glucopyranosyl-(1-->4)-beta-D-galactopyranoside], (25R)-3 beta-hydroxy-5 alpha spirostan-12-one (hecogenin) 3-O-[O-beta-D-glucopyranosyl-(1-->2)-O-[O-beta-D apiofuranosyl-(1- ->4)- beta-D-xylopyranosyl-(1-->3)]-O-beta-D-glucopyranosyl-(1- >4)-beta-D- galactopyranoside] and hecogenin 3-O-[O-beta-D-glucopyranosyl-(1-->2) O-[O-beta-D- apiofuranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->3)]-O-beta-D-gluc opyranosyl- (1-->4)-beta-D-galactopyranoside], respectively. The isolated saponins were examined for inhibitory activity using 12-O-tetradecanoylphorbor-13 acetate-stimulated 32P-incorporation into phospholipids of HeLa cells as the primary screening test to identify new antitumour-promoter compounds. PMID- 8729464 TI - Araliasaponins I-XI, triterpene saponins from the roots of Aralia decaisneana. AB - Seven new oleanane-type and four new ursane-type triterpene saponins, named araliasaponins I-XI were isolated from the roots of Aralia decaisneana, together with four known triterpene saponins. On the basis of the chemical and spectroscopic evidence, the structures of these new saponins were elucidated as follows: 3-O-beta-D-xylopyranosyl-(1-->3)-beta-D- glucopyranosyl-(1-->3)-[beta-D xylopyranosyl-(1-->2)]-alpha-L- arabinopyranosyl oleanolic acid 28-O-beta-D glucopyranosyl ester, 3-O-beta-D-glucopyranosyl-(1-->3)-alpha-L- arabinopyranosyl oleanolic acid 28-O-beta-D-glucopyranosyl-(1-->6)-beta-D- glucopyranosyl ester, 3 O-beta-D-glucopyranosyl-(1-->3)-[beta-D- xylopyranosyl-(1-->2)]-alpha-L arabinopyranosyl oleanolic acid 28-O-beta-D-glucopyranosyl-(1-->6)-beta-D glucopyranosyl ester, 3-O-beta-D-glucopyranosyl-(1-->3)-[beta-D-xylopyranosyl-(1- >2)]-beta-D- glucopyranosyl oleanolic acid 28-O-beta-D-glucopyranosyl ester, 3-O beta-D-glucopyranosyl-(1-->3)-[beta-D-xylopyranosyl-(1-->2)]-beta-D- galactopyranosyl oleanolic acid, 3-O-beta-D-glucopyranosyl-(1-->3)-[beta-D- xylopyranosyl-(1-->2)]-beta-D-galactopyranosyl oleanolic acid 28-O-beta-D glucopyranosyl ester, 3-O-beta-D-glucopyranosyl-(1-->3)-[beta-D-xylopyranosyl-(1- >2)]-beta-D- galactopyranosyl oleanolic acid 28-O-beta-D-glucopyranosyl-(1-->6) beta-D-glucopyranosyl ester, 3-O-beta-D-glucopyranosyl-(1-->3)-[beta-D xylopyranosyl-(1-->2)]- alpha-L-arabinopyranosyl ursolic acid 28-O-beta-D glucopyranosyl ester, 3-O-beta-D-glucopyranosyl-(1-->3)-[beta-D-xylopyranosyl-(1- >2)]-alpha-L - arabinopyranosyl ursolic acid, 3-O-beta-D-glucopyranosyl-(1-->3) alpha-L-arabinopyranosyl ursolic acid 28- O-beta-D-glucopyranosyl-(1-->6)-beta-D glucopyranosyl ester and 3-O-beta-D-glucopyranosyl-(1-->3)-[beta-D-xylopyranosyl (1-->2)]- beta-D-glucopyranosyl ursolic acid 28-O-beta-D-glucopyranosyl ester. PMID- 8729465 TI - Stoichiometric studies of tannin-protein co-precipitation. AB - Co-precipitation of a series of galloylglucoses (hydrolysable tannins) with bovine serum albumin (BSA) was studied stoichiometrically by analysing both galloylglucoses and BSA in the precipitates using HPLC. BSA-precipitating ability increased mainly with an increase in the number of galloyl groups in a galloylglucose molecule but was also affected by the position of the galloyl group (penta- > tetra- > 2,3,6-tri- > 2,3,4-tri- >> di- >> monogalloylglucose). The precipitated BSA increased linearly with an increase in the number of galloyl groups bound to a BSA molecule. BSA-precipitating abilities of the galloylglucoses were closely related to their relative affinities for BSA. These results suggest a two-stage mechanism: initial complexation of galloylglucose with BSA and subsequent precipitation, as a mechanism of the co-precipitation. PMID- 8729466 TI - [The serological properties of a lysine producer developing in a batch culture]. AB - Immune sera specific to bacterial cells at various development stages of periodic culture have been obtained with the purpose to study serological properties of lysin producer. Using the method of immunoenzyme analysis ELISA it is shown that there is certain dependence between the producer properties and the culture age. Different strains-producers used in production to-day are serologically close between each other. When using 49 strains-contaminants of lysin production (representatives of 10 genera) the absence of serological affinity between the producer and foreign microflora has been demonstrated. Preliminary data prove the prevalence of cells producers of thermostable antigens on the cell surface. PMID- 8729467 TI - [The phenotypic heterogeneity of meningococcal strains by their serogroup trait]. AB - It is shown that heterogeneity of meningococci of the outlined serogroups is formed due to appearance of nonagglutinable individuals in subpopulations but heterogeneity of polyagglutinable strains is characterized by incorporation not only of cross-reacting cells, but also microcommunities of each separate group which make a total feature. Serologically inactive strains either may be homogeneous or may form variants with low (to 20%) and high (to 44%) specific weight of agglutinable colonies. Strains in the centres embrace different concentrations of microcommunities which are identical by their serogroup properties to subpopulations of the meningitis pathogen. PMID- 8729468 TI - [Anti-idiotypic antibodies as vaccinal and diagnostic preparations in viral infections]. AB - The data concerning characteristics of anti-idiotypic antibodies and their use as vaccine and diagnostic preparations at virus infections have been reviewed. Main principles of the anti-idiotypic antibodies formation and functioning have been described. Methodical aspects of their production and testing as well as their positive and negative properties when used as vaccine were also generalized. PMID- 8729469 TI - Pulmonary oxygen uptake kinetics in nonsteady state. AB - The body does not always achieve a steady state and is more often kept in a nonsteady state. It is important not only in terms of physiological anthropology but also in its application to rehabilitation and training to clarify whether oxygen transport or utilization is the limiting factor in oxygen uptake in the nonsteady state. Hypotheses concerning the limiting factor in oxygen uptake have their own rationales, and it is still controversial as to which is actually responsible. The limiting factor in oxygen uptake may vary according to exercise conditions. Under certain conditions, oxygen transport would be responsible for limiting the oxygen uptake, and under others, oxygen utilization or both oxygen transport and utilization would be responsible. These conditions differ according to the type of exercise (e.g. step exercise vs. cycling, leg vs. arm exercise, and dynamic vs. static exercise), posture, recruited muscle fibers, and other experimental conditions. Considering that oxygen uptake is regulated by coupling among respiration, circulation, and metabolism, it may be possible that interactions among these functions vary in a complicated manner according to exercise conditions, which may require a different limiting factor in regulating oxygen uptake. PMID- 8729470 TI - Body size and composition in different somatotypes of Japanese college-aged women. AB - The purpose of this study was to determine differences in body size, composition and structure between three somatotypes of Japanese college-aged women. The study sample consisted of 30 sedentary female college students between 18 and 20 years of age. Ten subjects had an endomorphic ectomorph somatotype (mean weight 41.95 kg), 10 had an endomorph-ectomorph somatotype (mean weight 47.12 kg) and the remaining subjects had a mesomorphic endomorph somatotype (mean weight 55.37 kg). The mean heights for these groups did not differ significantly. The mesomorphic endomorph group had a higher gross weight and a higher percentage of all adipose variables than the other two groups, though these measurements were not significantly different between the endomorph-ectomorph and the endomorphic ectomorph groups. The mean lean body weight (LBW) for the mesomorphic endomorph group was significantly larger than that of the other two groups. The endomorph ectomorph group had the next largest LBW, and the mean for the endomorphic ectomorph group was significantly smaller than that of the other two groups. Inversely, mean values of LBW/weight(WT), LBW/total adipose tissue weight (TATW) ratio and total body water/WT were significantly lower for the mesomorphic endomorph group than for the other two groups. Furthermore, the ratios of internal adipose tissue to weight (IATW)/WT and subcutaneous adipose tissue to IATW (SATW)/IATW did not differ significantly between groups. In conclusion, college-aged women of different somatotypes within the same age and height range varied in body composition. Consequently, the notion of a small frame is incorrect. An accurate assessment of the human body's composition may provide a valuable insight into the ideal weight for optimal physiologic function. PMID- 8729471 TI - Peak oxygen uptake and respiratory function in persons with spinal cord injury. AB - This study investigated the association between peak oxygen uptake (peak VO2) during arm cranking exercise and respiratory function in paraplegics. Fourteen male paraplegics were recruited for the present study. The subjects were grouped according to the level of injury into the HL (Th3-Th8) and LL (Th11-L3) group. Prior to the maximal test, pulmonary function, including vital capacity (VC) and residual volume (RV), was measured in the sitting position. Mean peak VO2 in the LL group (1662 ml.min-1) was significantly greater than that in the HL group (1357 ml.min-1), corresponding to 82% of that in the LL group (P < or = 0.05). In respiratory function, the HL group showed marked restrictive impairment of ventilatory function. That is, VC and RV were significantly lower in the HL group than in the LL group (P < or = 0.05). The reduction in VC and RV is related to the degree of loss of control in respiratory functioning muscle mass. However, there was no clear-cut correlation between respiratory function and peak VO2 expressed as a function of body mass (ml.kg-1.min-1). In addition, a multiple linear regression analysis revealed that RV and VC were not associated with peak VO2 (ml.min-1) in contrast to the importance of body mass. It seems reasonable to conclude from these results that respiratory function is not an important factor in determining peak VO2 in the paraplegic. PMID- 8729472 TI - Effects of the thermal conditions of the dressing room and bathroom on physiological responses during bathing. AB - The effects of the thermal conditions of the dressing room and bathroom on the physiological responses during bathing were assessed. Six female students participated in this experiment. Three climate chambers were used as a living room, a dressing room and a bathroom. The living room was thermoneutral and maintained at 25 degrees C, while the thermal conditions of the dressing room and bathroom were as follows: (A) cold (10 degrees C), (B) cool (17.5 degrees C) thermoneutral (25 degrees C). The subjects wore standard clothing (0.65 clo). Heart rate (HR), blood pressure, rectal (Tre) and skin temperature, and subjective thermal sensation were recorded. 1) Marked increases in systolic blood pressure (SBP) after undressing and redressing in the dressing room and during washing were observed under the cold conditions. 2) A significant negative correlation was found between the dressing room temperature and increased SBP compared to before bathing (r = -0.684, p < 0.01, n = 18). 3) After exposure, mean skin temperature (Tsk) showed marked differences among the three conditions despite the rest taken under the same thermal conditions. 4) A significant negative correlation was found between Tsk and the increase in SBP of after undressing relative to that before bathing (r = -0.695 p < 0.01, n = 18). These findings suggested that 25 degrees C was the most appropriate temperature for the bathroom and dressing room, since the increase in blood pressure was minimum and subjective thermal sensation was neutral (neither cool nor warm) to warm under this thermal condition, and 17.5 degrees C at which the increase in blood pressure was within the physiological fluctuation range (+/- 10 mmHg) is the minimum tolerable temperature. PMID- 8729473 TI - The influence of location of electrode on muscle fiber conduction velocity and EMG power spectrum during voluntary isometric contraction measured with surface array electrodes. AB - A surface array electrode was used to investigate muscle fiber conduction velocity (MFCV) and EMG power spectrum during voluntary isometric contraction of m. biceps brachii. The mean power frequency (MPF) and the total power (TP) at various locations of the muscle were obtained from the power spectrum. MFCVs at various locations of the muscle were measured directly using the averaging method. The values of MPF, TP and MFCV were identified with respect to the electrode locations on the m. biceps brachii. MPF was shown as high near the end plate and low near the tendon of the muscle during contraction of 40% of isometric maximum voluntary contraction (MVC). TP showed low value near the end plate and the tendon of the muscle fiber and different values at different location setting electrode during voluntary isometric contraction. TP at each location on the muscle surface increased when the contraction levels increased in the contractions range of 20 to 60% MVC. MFCVs showed a high value near the end plate and the tendon of the muscle during the contraction of 40% MVC. These results indicated that MPF, TP and MFCV at different locations on the muscle were different along the length of the muscle fiber. PMID- 8729474 TI - Effects of two kinds of underwear on thermophysiological responses and clothing microclimate during 30 min walking and 60 min recovery in the cold. AB - Thermophysiological responses and clothing microclimate under the influences of different underwear materials were compared during walking and recovery in the cold. Two kinds of underwear were used: two layers of cotton underwear with two piece long-sleeved shirt and long-legged trousers (C), two layers of polypropylene underwear with two-piece long-sleeved shirt and long-legged trousers (P). In addition, the subject put on a two-piece ski suit of 100% polyester including 100% polyester padding. Seven adult females served as subjects in this study. The test was done in a climatic chamber at an ambient air temperature of 2 degrees C, a relative humidity of 65% and an air velocity of 0.14 m.s-1. The subject walked on a motor-driven treadmill with a 6 km/h speed for 30 min followed by 60 min recovery. Rectal temperature, skin temperatures, clothing microclimate (temperature, humidity), metabolic heat production and heart rate were measured. Furthermore, subjective ratings on thermal sensation, sweating/shivering sensation, clothing wettedness sensation and skin wettedness sensation for whole body were asked. The major findings are summarized as follows: 1) Mean skin temperature was not significantly different during walking, but it was significantly higher in P than in C during the recovery. 2) The absolute humidity of innermost layer and outermost layer were not significantly different during walking, but it was significantly higher in P than in C during the recovery. 3) Clothing microclimate temperature of innermost was not significantly different during the first half of walking, but it was significantly higher in C than in P during the second half of walking and significantly lower in C than in P during the recovery. Clothing microclimate temperature of outermost was not significantly different during walking, but it was significantly higher in P than in C during the recovery. 4) Metabolic heat production for the last 10 min during recovery tended to be higher in P. 5) The degree of skin wettedness sensation and clothing wettedness sensation for whole body was significantly higher in P during walking and recovery. Thus, it was concluded that two kinds of underwear with different properties to moisture could influence, not only clothing microclimate, but also physiological parameters like skin temperatures and metabolic heat production in the cold differently. PMID- 8729475 TI - Distribution of muscle fiber conduction velocity of M. biceps brachii during voluntary isometric contraction with use of surface array electrodes. AB - Surface electromyogram (EMG) was recorded in m. biceps brachii during the contractions of 20, 40 and 60% of maximum voluntary isometric contraction (MVC) in twelve healthy male subjects, using surface array electrodes. The distribution of muscle fiber conduction velocity (MFCV) was found directly using the averaging technique and the cross-correlation function technique. MFCVs in the region of 20 40 mm measured from end-plate denoted constant value of about 4 m/s in 20% MVC, while MFCVs in the region around end-plate and tendons showed about 10 m/s in 20% MVC. The values of MFCV depended on the contraction levels of muscle. The model for the generation of MFCV which considered the ensemble of muscle fibers with the shape of a cone was proposed. The theoretical values of MFCV by the muscle fiber ensemble model (MFE model) proposed in the paper showed in good agreement with the experimental results. PMID- 8729476 TI - Flow-induced changes in shape and cytoskeletal structure of vascular endothelial cells. AB - Changes in shape and cytoskeletal structure of vascular endothelial cells induced by fluid-imposed shear stress were studied in vivo and in vitro. In in vivo experiments, aortic tissue specimens including the flow dividers of the branching of the left subclavian artery and the aortic intercostal ostium were obtained and their endothelial cell shapes were observed using the scanning electron microscope. It was found that the shape and orientation of endothelial cells were strongly affected by blood flow conditions. In in vitro cell culture experiments, porcine aortic endothelial cells were cultured on glass coverslips and extracellular matrix coated (ECM coated) glass coverslips and exposed to shear stress using a parallel plate flow chamber. When we applied a shear stress of 2 Pa for 24 hrs, the rearrangement of F-actin filaments occurred within 3 hrs and preceded the cell shape change in the early stage after shear exposure. The endothelial cells on ECM-coated coverslips exhibited more elongated cell shapes even under no-flow conditions. After exposure to shear stress, the endothelial cells on ECM-coated glass showed more retarded elongation and orientation to the direction of flow than those on no-coated glass, suggesting that the anchorage to the substrate was enhanced by ECMs. PMID- 8729477 TI - Junction complexes of endothelial cells in atherosclerosis-prone and atherosclerosis-resistant regions on flow dividers of brachiocephalic bifurcations in the rabbit aorta. AB - To determine how hemodynamic forces influence the permeability of endothelial cells, we observed the composition of junction complexes of the endothelial cells at the flow dividers of bifurcations of the aorta in normolipidemic and hyperlipidemic rabbits by a freeze-fractured replica method. We found that the leading edge of the brachiocephalic flow dividers exposed to high shear stress in a laminar fashion was nonsudanophilic and was covered by long fusiform endothelial cells predominantly showing continuous tight junctions, rather than discontinuous forms, and smooth, regular gap junctions. In contrast, the "hip" of the brachiocephalic flow dividers, a region of relatively low shear stress, was sudanophilic, and was covered by ellipsoidal cells. The tight junctions were primarily discontinuous and the gap junctions had an irregular shape. These findings suggest that exposure of the arterial wall to a relatively low wall shear stress, rather than to high shear stress, may functionally activate the endothelial cells, increasing intercellular permeability. These events may increase the vulnerability of these regions of the vessel to atherosclerosis. PMID- 8729478 TI - Effects of fluid shear stress on bone cells. AB - Neonatal rat calvarial cells (osteoblast-enriched) were cultured in monolayer on glass slides and subjected to a step increase in shear stress ranging from 3 to 60 dyne/cm2 in a laminar flow chamber. The level of intracellular free calcium ion concentration was monitored using Fura-2 fluorescence during the application of stress. Application of a step increase in stress resulted in a transient increase in calcium starting about 10-20 s after turning on the pump, reaching a peak in about 35 s and declining thereafter, despite the continued shear stress, and reaching near baseline values in about 100 s. No change in cytoplasmic calcium was observed with a step increase in shear stress of 3 dyne/cm2. Between 6 dyne/cm2 and 60 dyne/cm2, the magnitude of the calcium response increased with the applied shear stress. PMID- 8729479 TI - Changes in the rheological properties of the cell wall of plant seedlings under simulated microgravity conditions. AB - In this review article, we discuss the changes in the mechanical properties of the primary cell wall of different organs of several species of plant seedlings grown under simulated microgravity conditions. We compared growth and tropistic responses of these organs growing under different microgravity conditions, namely: 1) 3-D clinostat (three-dimensional clinostat with two axes); 2) water submergence for rice seedlings, and 3) for comparison, an accelerated gravity using a specifically designed centrifuge. We measured the minimum stress relaxation time as the parameter representing the mechanical property of the cell wall. We also measured extensiblity, in mm/g. The 3-D clinostat condition disturbed the normal gravitropic response of organs but affected growth rate and mechanical properties of the cell wall very little. Water-submergence of rice seedlings caused an acceleration of coleoptile elongation in the dark and caused a marked change in the mechanical property of the cell wall. However, the additional gravity of 30-135 xg showed only a small effect on growth and the mechanical property of the cell wall. PMID- 8729480 TI - Non-Newtonian viscosity of human blood: flow-induced changes in microstructure. AB - Flow-induced changes in the red cell microstructure of human blood are identified from mechanical and optical evidence. On initiation of steady flow, a new microstructure develops as the shear strain increases through unit strain. This structure is identified with the formation of layers of red cells that slide on plasma layers (Thurston, 1989). At low shear rates, the cell layers are composed of aggregated cells, but at higher shear rates, the aggregates degrade to form thinner layers of oriented, compacted cells. The viscosity is determined by the hematocrit, the degree of compaction and viscosity within the cell layers, and the plasma viscosity. Degradation of cell aggregates is controlled by 1) the time required for the strain to increase by one unit (delta t1 = 1/shear rate) and 2) the dominant viscoelastic relaxation times of the red cell structures. Structures having relaxation times > delta t1 are degraded by cell disaggregation; when delta t1 is less than the shortest relaxation time of the layered cells, disaggregation and (cell and plasma) layer formation are nearly complete. Analyses of the non-Newtonian viscosity and cell layer characteristics are given for both normal and hardened cells. PMID- 8729481 TI - Influence of hematocrit on erythrocyte aggregation kinetics for suspensions of red blood cells in autologous plasma. AB - The influence of hematocrit on red blood cell aggregation was investigated using a modified automated Myrenne Aggregometer System. Hematocrit values ranged from 20-70% at 10% increments, and were produced by resuspending washed red cells in autologous plasma. The Myrenne System produced seven kinetic parameters of red cell aggregation. The results indicate that hematocrit (Hct) had a strong and nonlinear effect on erythrocyte aggregability. At both low and high hematocrit, the aggregation intensity (AI) tends to decrease, reaching a plateau at 40% hematocrit. The disaggregation shear rate demonstrates an exponential relationship as In gamma TMIN = 3.29-0.0837 Hct. Time constants of the aggregation process such as T1/2, TSLOW and TFAST were rather inconsistent. The increase of hematocrit increased the rate of the fast phase aggregation and slightly slowed the secondary aggregation. PMID- 8729482 TI - Effects of calcium permeabilization on RBC rheologic behavior. AB - Although many RBC rheological properties have been previously described in detail, the biochemical mechanisms leading to premature destruction of red blood cells are less clear. However, several biochemical processes have been suggested as possible mechanisms for membrane structural alterations (e.g., crosslinking of membrane proteins, oxidant damage, binding of cytoplasmic proteins, and altered intracellular ion composition). We have carried out a series of studies aimed at evaluating the effects of calcium-regulated membrane-bound hemoglobin (Hbm) on RBC and derived ghost rheologic behavior. Intracellular calcium was elevated by 10 microM A23187, with cell deformability determined via the Cell Transit Analyzer (CTA). Our results indicate: 1) Linear, positive correlations between Hbm and average RBC rigidity and 2) a marked influence of heterogeneous calcium concentration on both Hbm and rheologic properties for various subpopulation. These findings therefore suggest the importance of hemoglobin-membrane interactions as a determinant of erythrocyte deformability, and may be relevant to RBC aging as well as to diseases such as sickle cell anemia, hereditary spherocytosis and thalassemia. PMID- 8729483 TI - Hydrodynamic properties of nucleosomal fibers and chromatin higher-order structure. AB - We studied the comparative hydrodynamic properties of chromatins differing in transcriptional activity and in linker DNA length. Chromatins were prepared from the nuclei of pigeon brain cortical neurones, rat thymus and sea urchin sperm. The sizes of linker DNA are 20, 50 and 100 base pairs (b.p.), respectively. Oligonucleosomes of different chain length were isolated from micrococcal nuclease-digested nuclei and studied by the method of sedimentation velocity. Rheological behavior of nucleosome oligomers depends on the number of nucleosomes in the chain and on the ionic strength in the range between 5 and 85 mM was analyzed on the basis of a cylinder model using different hydrodynamic theories. It is shown that hydrodynamic properties of oligonucleosomes from all types of investigated chromatins at low ionic strength can be well described by the model of a three-dimensional zig-zag chain with a diameter of about 20 nm, the DNA packing ratio growing with the increase of linker DNA length. Increasing ionic strength induces condensation of the zig-zag-shaped chain of nucleosomes into a two-start double superhelix with densely packed nucleosomes. Linkers are arranged inside the superhelix, which is bent or folded into a loop, depending on its size. PMID- 8729484 TI - Viscoelasticity of hyaluronic acid with different molecular weights. AB - Storage and loss moduli of hyaluronic acid solutions with different molecular weights were observed as a function of frequency in the presence of sugars and salts. The hyaluronic acid solutions of higher molecular weight (Mw > 17 x 10(6)) fractions showed entanglement, whereas lower molecular weight fractions did not. For inducing the entanglement of molecular chains of hyaluronic acid, increasing the molecular weight of hyaluronic acid was more effective than increasing the concentration of the lower molecular weight fractions (Mw = 7. 8 x 10(5). Glucose, fructose, galactose, and sucrose increased both storage and loss moduli, while NaCl, KCl and CaCl2 decreased both moduli. It is suggested that sugars create hydrogen bonds and strengthen the transient network. Cations shield the electrostatic repulsion of hyaluronic acid molecules, and the polymer chains are shrunk into compact coils from expanded stiffened coils. PMID- 8729485 TI - Determination of material constants and hydraulic strengthening of trabecular bone through an orthotropic structural model. AB - A statistical mathematical model is used to describe the viscoelastic behavior of trabecular bone from the description of its microstructure. Trabecular bone is regarded as an orthotropic composite material consisting of two phases: The trabeculae, modelled as a non-linear elastic material, and the marrow, modelled as a Maxwell element. The macroscopic constitutive equation contains, as parameters, the constants of the material phases' constitutive equations, the respective volume fractions and parameters descriptive of the geometry of trabecular bone. With this model, the contribution of the marrow to the mechanical behavior of trabecular bone is determined and it is calculated that, under a step load and at strain rate, 10 min-1, the marrow can bear about 25% of the applied load. The constants of the material phases are also calculated from a series of stress-relaxation curves on trabecular bone specimens. It is concluded that the trabeculae display non-linear elastic behavior. PMID- 8729486 TI - The significance of red cell surface area to the permeability of fibrin network. AB - Thrombus formed under low shear conditions in the intravascular compartment consists of red cells enmeshed within a fibrin network framework. The permeability of the network determines the rate of molecular transport by convection. The effect of red cells on the permeability of the fibrin network was examined in networks developed from red cell suspensions in platelet-rich (PRP) and platelet-poor plasma (PPP). Red cell-rich networks developed in PRP were significantly more permeable than those developed in PPP. Network permeability decreased linearly with increase in hematocrit due to volume exclusion and cell surface hydraulic drag. The hydraulic resistance of the red cells was similar in order of magnitude to that of the fibrin fibers. The hydraulic resistance was calculated to be dependent on the surface area of the red cells, as well as on red cell concentration. Calculation of red cell surface area from permeability (45-90 microns2) was found to be lower than the known surface area (145 microns2). From these studies, it is suggested that red cells entrapped within the network are aggregated. Aggregation is promoted by low shear plasma conditions during fibrin polymerization and by fibrin polymerization intermediaries. The degree of red cell aggregation regulates the hydraulic resistivity of the red cells, and the fibrin fiber structure regulates the hydraulic resistivity of the fibrin network. Both are significant determinants of the network permeability and, therefore, of molecular transport in thrombi. PMID- 8729487 TI - A new laser photometric technique for the measurement of erythrocyte aggregation and sedimentation kinetics. AB - The aggregation kinetics of erythrocytes is studied by using laser light scattering. Laser light is passed through a well mixed sample of blood and the forward-scattered light intensity is recorded continuously. During the process of aggregation and sedimentation under a gravitational field, erythrocytes are subjected to different orientations. The events of a single cell's movement, formation of aggregates and their sedimentation produce a variation in scattered light intensity. The mean intensity variation that bears this information is effectively employed to arrive at the kinetics of single cells, one-dimensional aggregates and three-dimensional aggregates during sedimentation. In addition to this, the time intervals of different phases of aggregation and sedimentation are also measured. All these parameters are obtained for erythrocytes suspended in native plasma. Then, the results are compared with erythrocytes suspended in dextran (Dx) 70 and Dx 500. PMID- 8729488 TI - Effect of electric field on erythrocyte sedimentation rate. IV. Volume fraction dependence of human red cells in plasma. AB - We measured the volume fraction dependence of erythrocyte sedimentation for human red cells in autologous plasma while the volume fraction, Hct, ranges from 0.220 to 0.650. A modified electrophoretic instrument was employed to investigate the electric effect of an upward DC field on erythrocyte sedimentation at a constant source supply of Vs = 60.0 +/- 0.5 V. The sedimentation curves all coincided well with the sigmoidal type equation, l = lmax/?l + (t50/t)n?, at Vs = 0, or 60.0 V. Here, lmax is the length of plasma level, l, when the time, t, tends to infinity; t50 is the elapsed time when the plasma level falls to lmax/2, and n is a constant (n > 0). A non-linear least-squares method was employed to fit experimental data. It was found that lmax decreased and t50 increased with the increase of Hct, while n ranged from 1.53 to 3.95. The volume fraction dependence of sedimentation velocity, Vmax, defined as the maximum slope of the sedimentation curve, fits well with an exponential-type curve, both at Vs = 60.0 V and at Vs = 0 V: Vmax = Vs (1-Hct)exp(-BHct), where Vs is the velocity at infinite dilution and B is a phenomenological parameter. In the range of Hct = 0.220-0.650, Vs and B are 326.1 mm/hr and 4.30, respectively, at Vs = 60.0 V, while they are 130.4 mm/hr and 2.46, respectively, at Vs = 0 V. PMID- 8729489 TI - Heparin effect on red blood cell aggregation. AB - We studied the effect of relatively high dosages of heparin (25-150 U/ml) on some rheological parameters of donor blood. It was found that heparin increased the erythrocyte sedimentation rate (ESR) and low shear blood viscosity at all concentrations studied. The increase of these parameters indicates an elevation in RBC aggregation. In all samples with a heparin concentration of 100 U/ml, the average ESR was increased by approximately 75% (p < 0.001); whole blood viscosity increased under shear rate, gamma = 0.277 sec-1, by approximately 30% (p < 0.001). Since bolus injection of heparin may result in transient high local concentration, these results suggest a possible additional adverse effect to the well known heparin effect on platelet aggregation. PMID- 8729490 TI - Unilateral vocal fold paralysis: a review of the current methods of surgical rehabilitation. AB - This review article discusses the surgical treatment of patients suffering from unilateral vocal fold paralysis who have already been assessed and considered appropriate candidates for surgery. There are currently three main methods of surgical rehabilitation; injection medialisation; laryngeal framework surgery; re innervation procedures. PMID- 8729491 TI - Client centred hypnotherapy in the management of tinnitus--is it better than counselling? AB - The aim of this study was to assess whether client centred hypnotherapy (CCH) which required three sessions with a trained therapist was superior to a single counselling session in reducing the impact of tinnitus. Patients were randomly allocated to receive either counselling (n = 42) or CCH (n = 44). The outcome measures were: tinnitus loudness match, subjective tinnitus symptom severity score, trend of linear analogue scale, request for further therapy and whether the patient had an impression of improvement in their tinnitus after treatment. CCH was no better than counselling in reducing the impact of tinnitus using the three quantative measures of tinnitus, and requests for further follow up. The only significant difference between the two therapies was that 20 (45.5 per cent) of the CCH group reported a general sense of improvement compared to six (14.3 per cent) in the counselling group, this is significant p < 0.01. The study did not demonstrate whether this was a genuine hypnotic effect or simply a response to the additional attention from the therapist. PMID- 8729492 TI - Hearing disability before and after radiotherapy for nasopharyngeal carcinoma. AB - This paper evaluates post-irradiation hearing changes in patients with nasopharyngeal carcinoma (NPC) from a disability orientated approach, which takes into account binaural hearing. Newly diagnosed patients with NPC were studied before radiotherapy, and at four to 12 months (mean 9.2 months) after radiotherapy, provided they remained disease-free. Each patient was examined clinically and with pure tone audiograms. Tympanometry was used to confirm middle ear effusion. Averaged hearing thresholds over 0.5, 1 and 2 kHz were evaluated. If abnormal (> 30 dB), the resultant hearing disability was illustrated by a modified Glasgow Plot. Twenty-three males and 10 females completed the study. Middle ear effusions resulted in 39.3 per cent (binaural in two patients) and 33.3 per cent (binaural in five patients) of patients having hearing disability pre- and post-irradiation respectively. No patient had hearing disability as a result of a sensorineural loss. It is recommended that future reporting of post irradiation hearing changes in patients with NPC, as in middle ear surgery, be considered from a disability-orientated approach. PMID- 8729493 TI - Laser-Doppler blood flowmetry measurement of nasal mucosa blood flow after injection of the greater palatine canal. AB - Laser-Doppler blood flowmetry was used to measure the mucosal blood flow of the inferior turbinate before and after injection of the greater palatine canal with 2 ml of a 0.5 per cent bupivicaine hydrochloride solution. Injection of the greater palatine canal is a useful technique in the control of posterior epistaxis. The arterial blood supply to the inferior turbinate is via a single descending branch of the sphenopalatine artery. We conjectured that injection of the pterygopalatine fossa, via the greater palatine canal, would result in a reduction of blood flow to the inferior turbinate. In this study injection of the pterygopalatine fossa caused only a 4.7 per cent decrease in blood flow to the inferior turbinate mucosa (p = 0.571). Elevation of the head by an angle of 20 degrees reduced nasal mucosal blood flow by 38.3 per cent (p < 0.0001). Depression of the head by an angle of 20 degrees increased nasal mucosal blood flow by 74.7 per cent (p < 0.0001). We conclude that there is an adequate collateral blood circulation to the anterior portion of the inferior turbinate. PMID- 8729494 TI - Septoplasty as a day-case procedure--a two centre study. AB - Day-stay surgery is an integral part of Otolaryngology. Many procedures which have traditionally been thought to require overnight stay can be carried out on a day care basis. We report our experience of treating 163 patients admitted for septal surgery as a day-case procedure. The paper summarizes the experience of two centres. One of these is a London Teaching Hospital, where surgery was performed through a dedicated day-case unit, and the other is a District General Hospital where patients were admitted to a day-case unit but had their surgery on a routine in-patient list. As the result of this study we conclude that day-stay septoplasty is associated with a low complication rate and is a safe and acceptable procedure provided that strict selection criteria are followed. The cost implications are discussed. PMID- 8729495 TI - The treatment of hyposmia with intranasal steroids. AB - Hyposmia is a neglected symptom in patients with rhinitis. We studied 25 patients presenting with perennial rhinitis. Fifteen patients expressed hyposmia as a significant symptom. University of Pennsylvania smell identification test (UPSIT) and visual analogue scales (VAS) were used to score the symptoms of hyposmia, nasal obstruction and nasal discharge before and after six weeks treatment with betamethasone sodium phosphate drops. Those patients with initial symptoms of hyposmia significantly improved their UPSIT scores (p = 0.00009) and their VAS scores for hyposmia (p = 0.00133). Despite a significant decrease in the sensation of nasal obstruction, the non-hyposmics showed no increase in UPSIT scores after betamethasone therapy. There was no clear correlation between UPSIT results and other symptom scores. The judicious use of betamethasone drops in the treatment of rhinogenic hyposmia can be recommended. PMID- 8729496 TI - Complications of paediatric tonsillectomy post-discharge. AB - A post-operative survey of 291 children was conducted to assess morbidity found at home in the first five days post-tonsillectomy. Analgesic requirement (92.4 per cent), signs of distress (90.4 per cent), otalgia (69.1 per cent), halitosis (66.7 per cent) were common. Physical or behavioural changes (36.8 per cent) and secondary haemorrhage (8.9 per cent) were also prominent features. Nausea was reported in 59 children (20.3 per cent) and delayed return to a normal diet which in turn predisposed to secondary haemorrhage. Return to normal diet was independent of pain. General Practitioners were consulted by 60.6 per cent and more than half were prescribed drugs. The presence of an organized clot in the tonsillar fossa (3.45 per cent) post-operatively did not delay discharge from hospital and none of these patients re-presented with secondary haemorrhage. Based on their child's experience in this study, only one third of the parents approved of day-case tonsillectomy in principle, a finding which has implications for the instigation of day-case tonsillectomy procedures. PMID- 8729497 TI - The role of stroboscopy in the management of a patient with a unilateral vocal fold paralysis. AB - Stroboscopy is well established as an essential diagnostic tool in the assessment of the vocal folds during phonation. This paper analyses the stroboscopic findings in 100 patients with a unilateral vocal fold paralysis. Reliable stroboscopic signals were only obtained in patients with the paralysed fold close to the midline. These patients seldom require surgery however, usually responding to speech therapy with laryngeal compensation giving a good voice. Most patients that require surgery have a large glottal deficiency, but in this series these patients did not give an adequate signal for analysis. Although useful in the assessment of the muscle tone of the paralysed fold, the influence of stroboscopy on the surgical treatment in this series was limited. PMID- 8729498 TI - The management of dysphagia in jugular foramen surgery. AB - From 1985-1994, the Skull Base Unit at St. Vincent's Hospital, Sydney, operated on 61 patients with tumours involving the jugular foramen. Pre-operative assessment by a Speech Pathologist and the institution of swallowing techniques prior to surgery have improved post-operative morbidity. Ancillary procedures at the time of surgery were not required in the majority of cases. An individual assessment of each patient early in the postoperative period was found to be more important with regard to the benefits of supplementary surgery. The majority of patients with dysphagia settled with conservative management and only a few underwent ancillary surgery. It is perceived that the cortical and subcortical control of swallowing is a major factor in the rehabilitation of these patients. PMID- 8729499 TI - The deep parotid lymph nodes: an anatomical and oncological study. AB - In order to better define the anatomical and clinical importance of the deep parotid lymph nodes, the surgical specimens obtained from a series of 18 total parotidectomies were evaluated. In 10 cases primary parotid pathology was found, whereas in the remaining eight cases metastases to glandular lymph nodes were present. One hundred and forty-nine lymph nodes, in all, were identified (range 3 14, average 8.2, mean 8), 118 located in the superficial parotid lobe (range 3 11, average 6.5, mean 7), and 31 in the deep lobe (range 0-5, average 1.7, mean 2). In the group of patients with oncological pathology, of the total 64 lymph nodes examined, 21 were found to be sites of metastasis, 11 in the superficial lobe, and 10 in the deep lobe. The above findings confirm the anatomical and oncological importance of the deep parotid lymph nodes, and highlight the necessity of a total parotidectomy in all cases in which intraglandular spread of lymph node metastases is certain or suspect. PMID- 8729500 TI - Primary neuroblastoma of the facial nerve presenting as a recurrent facial paralysis. AB - A case is described of a primary neuroblastoma of the facial nerve in a 13-year old boy presenting with a recurrent facial paralysis. This tumour was excised preserving the nerve and followed with post-operative radiotherapy. The pathology of the tumour is described and facial nerve tumours briefly discussed as a cause of facial palsy. There have been no other cases of a primary neuroblastoma of the facial nerve arising at this site reported in the last 20 years. PMID- 8729501 TI - Relapsing polychondritis presented as inner ear involvement. AB - We report a rare case of relapsing polychondritis with an initial symptom of inner ear involvement. This 53-year-old Japanese man experienced a hearing difficulty, tinnitus in both ears, and dizziness of sudden onset, but lacked auricular chondritis at that time, which is the most frequent finding in relapsing polychondritis. Thus it was difficult to reach a correct diagnosis. Steroid therapy, with oral prednisolone 15 mg daily, was effective. Almost two months after we began the steroid therapy, the patient complained of losing interest in his work and reported a hallucination vision on the TV screen, so the dose of prednisolone was decreased to 10 mg. The hallucinations then disappeared, but the serum level of C-reactive protein increased highly. To reduce the dose of prednisolone, we tried low-dose oral methotrexate. However, we had to discontinue it when the patient experienced severe vomiting and diarrhoea. As adjuvant therapy, we then administered Sho-saiko-to, Chinese herbal medicines with few side effects. Symptoms and laboratory abnormalities then improved markedly. PMID- 8729502 TI - Osteoma of the internal auditory canal: case report. AB - A 42-year-old man with normal hearing presented with a long history of vertigo and tinnitus. CT scan showed large osteomata of the internal auditory canal. Magnetic resonance imaging (MRI) was normal. The osteomata were removed surgically via the retrosigmoid approach and examined histologically. His symptoms were abolished. As MRI has become the gold standard in the search for small acoustic tumours it is likely that symptomatic bony lesions, rare though they are, will be missed if MRI is the sole imaging modality. A review of the literature is included. PMID- 8729503 TI - Non-Hodgkin's lymphoma presenting as an isolated temporal soft tissue swelling. AB - Non-Hodgkin's lymphoma (NHL) of the head and neck has been described in several sites including the larynx, the pharynx, Waldeyer's ring, the nose and paranasal sinuses. NHL of the temporal region has not been previously described. We describe a 65-year-old woman with a diffuse swelling of the right temporal region. A histological diagnosis of NHL was made following complete excision of the lesion. No further treatment was given and there was no evidence of recurrence at six months. PMID- 8729504 TI - Primary malignant melanoma of the palatine tonsil: a case report. AB - The authors report a clinical case of a primary malignant melanoma of the right palatine tonsil in a 75-year-old woman. PMID- 8729505 TI - Adenomatoid hyperplasia of palatal minor salivary glands. AB - Adenomatoid hyperplasia of palatal minor mucous glands is rare but significant because the clinical appearance mimics malignant disease. The typical history of a painless, indolent palatal swelling, together with the histological picture of benign glandular hyperplasia and hypertrophy, are illustrated in this report. PMID- 8729506 TI - Schwannoma of the posterior pharyngeal wall: a case report. AB - Schwannoma arising in the posterior pharyngeal wall is rare. We report on a 60 year-old man who complained of discomfort in his pharynx, from whom a tumour was excised via an intraoral approach. No recurrence was seen after an 11-year follow up. The nerve origin of the tumour is most likely to be the peripharyngeal plexus. This is the third such case reported. PMID- 8729507 TI - Metastatic masseter muscle tumour: a report of a case. AB - Both primary and metastatic malignancies of the masseter muscle are rare. We report a case of metastatic renal cell carcinoma to the masseter muscle. It was incidentally found as a hypervascular mass in carotid angiography for delineating a recurrent metastatic brain tumour. Prior to surgical removal, intravascular embolization via the left facial artery was performed in order to decrease intra operative bleeding. The tumour was removed with minimum damage to the muscle fibres by the extraoral method, followed by a transient lower lip palsy. Metastatic intramuscular tumours, which are assumed to be due to haematogenous spread, are generally a sign of poor prognosis. PMID- 8729508 TI - Mediastinitis-a rare complication of a peritonsillar abscess. AB - We report a case of mediastinitis, in an otherwise healthy 25-year-old man, resulting from a peritonsillar abscess with extension through the parapharyngeal and retropharyngeal spaces. In our case the patient was primarily treated with needle aspiration, a method described in many publications as a safe alternative to incision and drainage. We emphasize that for peritonsillar abscesses a tonsillectomy or wide incision and drainage, instead of needle aspiration, might prevent the extension of the condition, thus preventing serious complications. PMID- 8729509 TI - Primary lingual tuberculosis: a case report. AB - A case of primary tuberculosis of the tongue in a 45-year-old male patient is presented. The clinical manifestation, diagnosis and the response to the antituberculosis treatment are considered. The previous literature is reviewed. PMID- 8729510 TI - Thrombosed jugular vein presenting as a hard neck mass. AB - We report an unusual case of thrombosis of a dilated external jugular vein presenting as a hard neck mass of sudden onset. The lesion's presentation, investigation, treatment and pathology are discussed. PMID- 8729511 TI - Subcutaneous diffuse neurofibroma of the neck: a case report. AB - A case of a rare and unusual variant of neurofibroma, diffuse neurofibroma (paraneurofibroma), in a young patient is presented. The clinical, radiological and histopathological features of this case are reported. The magnetic resonance imaging (MRI) features of the diffuse neurofibroma are comparable with those described in other neurofibromas. PMID- 8729512 TI - Post-stapedectomy reparative granuloma: a misnomer. AB - The pathophysiology of so-called 'reparative granuloma' occurring after stapedectomy has not been determined and universally accepted management of this rare complication has not yet been established. A case is presented in which a mass developed in the middle ear after the use of a fat/wire prosthesis in a stapedectomy. Histological assessment revealed nonspecific granulation tissue and fat necrosis. It is suggested that 'reparative granuloma' is a misnomer as there is no microscopic evidence in this case nor in the literature of granulomatous formation. The condition may follow stapedectomy or stapedotomy. Furthermore, the name leads to confusion with a different condition, giant cell reparative granuloma, which involves the jaws and rarely the temporal bone. An alternative name, 'Stapes surgery induced granulation tissue' (SSIG) is therefore suggested for this condition. PMID- 8729513 TI - Atypical intramuscular lipoma of the tongue. AB - Lipomatous tumours of the tongue are rare and most of them are benign ordinary lipomas. In this report, we describe the first case of atypical lipoma of the tongue in a 78-year-old male, and review the current literature and controversies surrounding the nomenclature and biology of this unusual lesion. PMID- 8729514 TI - Rectal carcinoma metastatic to the thyroid gland. AB - Clinically evident metastases to the thyroid gland are rarely found antemortem. A case of a 59-year-old woman with a history of rectal carcinoma, who presented with low back pain and a mass in the right lobe of her thyroid gland, is presented. The tumour of the thyroid was found to be metastatic adenocarcinoma from her previous rectal cancer. Other synchronous metastases were noted in her lumbar spine and kidneys. The clinical finding of metastases to the thyroid gland is rare, particularly from a colorectal primary. One must consider, however, the possibility of a tumour of the thyroid gland representing a secondary malignancy in any patient with a prior history of cancer. PMID- 8729515 TI - Paraganglioma: an unusual tumour of the parathyroid gland. AB - A case of paraganglioma arising within a parathyroid gland is reported. The lesion was an incidental finding in a block dissection of neck performed for squamous carcinoma of the pharynx. A well-circumscribed lesion, exhibiting the characteristic pathological features of a paraganglioma, was embedded within the right inferior parathyroid gland. Due to its location, the chief histological differential diagnosis was an unusual variant of parathyroid adenoma. Immunohistochemistry and electron microscopy assisted in reaching a diagnosis. This, as far as we are aware, is the first reported case of a paraganglioma of the parathyroid gland. PMID- 8729516 TI - KTP-532 laser tonsillectomy--a potential day-case procedure? PMID- 8729517 TI - Palatal myoclonus affected by neck posture. PMID- 8729518 TI - Investigation of the solitary thyroid nodule. PMID- 8729519 TI - Somatic mutations in the RET proto-oncogene in sporadic medullary thyroid carcinoma. AB - OBJECTIVE: We have determined the frequency of specific mutations in the RET proto-oncogene in sporadic medullary thyroid carcinomas (MTCs) and correlated the presence or absence of a codon 918 mutation with the clinical characteristics of these tumours. DESIGN: Thirty paraffin-embedded sporadic MTCs and two frozen MTCs were collected for analysis of specific mutations in the RET proto-oncogene in codons 609, 611, 618 and 620 (exon 10); 630 and 634 (exon 11); 768 (exon 13); 883 (exon 15) and 918 (exon 16). A novel primer was designed which introduced a restriction site for Rsal in the presence of the specific codon 918 mutation (ATG ->ACG) in these tumour samples. A 'clinical-genetic' correlation was performed comparing the presence of absence of the codon 918 mutation with the following clinical characteristics: age at diagnosis, tumour size, presence or absence of metastases, MTC related morbidity, and base line calcitonin levels at diagnosis or most recent follow-up. PATIENTS: Patients were classified as having sporadic MTC if there was no family history of C-cell hyperplasia, MTC, phaeochromocytoma or parathyroid disease. Retrospective review of patient records enabled complete clinical data to be obtained in 28 of 32 patients. MEASUREMENTS: Base line calcitonin levels were measured by radioimmunoassay or calcitonin enzyme linked immunoassay. Cysteine codons in exons 10 and 11, specifically codons 609, 611, 618, 620, 630 and 634, were screened for the presence of mutations by sequence analysis. Specific mutations occurring at codons 768, 883 and 918 were screened for by restriction endonuclease digestion of PCR products. RESULTS: The mutation at codon 918ATG-->ACG was found in 21 of 32 (66%) MTCs and the mutation at codon 883GCT-->TTT was found in one of 32 MTCs. Where possible, the presence of 'germline-type' mutations in codons 609, 611, 618, 620, 630 and 634 were excluded. Ten MTCs did not have a mutation in codons 768, 883 or 918 of the RET proto-oncogene. The presence or absence of the somatic mutation at codon 918 did not correlate with any of the above clinical characteristics. CONCLUSION: Somatic mutations in the RET protooncogene occur frequently in sporadic MTCs. PMID- 8729520 TI - Heterogeneous distribution of thyrotrophin receptor messenger ribonucleic acid (TSH-R mRNA) in papillary thyroid carcinomas detected by in situ hybridization. AB - OBJECTIVE: TSH suppression therapy has been reported to be effective in some cases of papillary carcinoma of the thyroid. However, there has been little information regarding the status of expression of TSH-R mRNA in papillary carcinomas. Therefore, we examined the expression of TSH receptor (TSH-R) mRNA in thyroid tumours by in situ hybridization. DESIGN: Surgically resected thyroid tumour specimens were used. The expression level of TSH-R mRNA was detected by in situ hybridization, and the difference between tumours and their adjacent normal thyroid tissues was assessed. PATIENTS: Tumours and adjacent normal thyroid tissues were obtained from nine patients with differentiated papillary carcinomas and eight with adenomas. MEASUREMENTS: We performed in situ hybridization on frozen sections of thyroid tissues. Digoxigenin-labelled oligonucleotide was used to hybridize and to detect TSH-R mRNA. We counted positively stained cells and non-stained cells in each section, calculated the positivity of stained cells and assessed the distribution of positively stained cells. RESULTS: In normal thyroid tissue, the positivity was 95.3 +/- 2.3% (mean +/- SD) and the distribution of TSH-R mRNA was homogeneous. In adenoma, positivity was 93.5 +/- 4.4%. In most cases of adenoma, the distribution was homogeneous. In papillary carcinoma, the expression level of TSH-R mRNA was significantly lower and the positivity was 81.9 +/- 12.6%. Furthermore, in five cases of papillary carcinoma, the distribution was heterogeneous; four of these cases were stage 3, while the remaining case was stage 1. Three of the four cases with a homogeneous distribution were stage 1, while one case was stage 3. CONCLUSIONS: The heterogeneity of TSH-R mRNA distribution suggests the effect of TSH on carcinoma cells may be variable. Furthermore, papillary carcinomas of advanced clinical stage showed a tendency to low expression and heterogeneous distribution of TSH-R mRNA. PMID- 8729521 TI - Polycystic ovaries in pre and post-menopausal women. AB - OBJECTIVE: Polycystic ovaries have been diagnosed in more than 20% of premenopausal women using ultrasound. The aim of this study was to determine whether polycystic ovaries exist in post-menopausal women. DESIGN: Two groups of women were studied; group 1 consisted of 18 post-menopausal volunteers and group 2 comprised 142 women, 94 of whom were post-menopausal who had recently undergone coronary angiography. MEASUREMENTS: Transabdominal and transvaginal ultrasound scans were performed and measurements made of uterine area, endometrial thickness and ovarian volume. The morphological appearance of the ovaries was also noted. Fasting blood samples were taken. Medical and menstrual questionnaires were completed. RESULTS: Polycystic ovaries were found in 8/18 (44%) of group 1 and 60/142 (42%) in group 2. Polycystic ovaries were detected in 35/94 (37%) of the post-menopausal women in group 2. Post-menopausal women with polycystic ovaries had larger ovaries containing more follicles compared with post-menopausal women with normal ovaries. Post-menopausal women with polycystic ovaries had higher serum concentrations of testosterone and triglycerides than had post-menopausal women with normal ovaries. CONCLUSIONS: Polycystic ovaries can be detected in post-menopausal women and have some of the same endocrine abnormalities which are evident in premenopausal women with polycystic ovaries, that is, raised serum concentrations of testosterone and triglycerides. PMID- 8729522 TI - Dyslipidaemia is associated with insulin resistance in women with polycystic ovaries. AB - OBJECTIVE: Polycystic ovary syndrome (PCOS) is characterized by hyperinsulinaemia and insulin resistance. Previous reports of lipid abnormalities in the syndrome have produced conflicting results which may, in part, be related to the lack of appropriate controls for the obese women with PCOS. Only one study has related lipid levels to insulin sensitivity. The objective of this study was to assess lipids and lipoproteins in women with PCOS, to compare the results with weight matched controls, and to relate the findings to indices of insulin secretion and action, and to menstrual history. DESIGN: A cross-sectional study of insulin sensitivity and lipids in a cohort of PCO subjects compared to weight and ethnic group matched controls. PATIENTS AND METHODS: We have therefore investigated glucose tolerance, plasma lipids and lipoproteins in 19 lean (LP) and 55 obese (OP) patients with PCO and compared the results with those in 22 lean (LC) and 15 obese (OC) control women. Insulin sensitivity was measured in the same subjects with a short insulin (0.05 U/kg i.v. insulin) tolerance test (LP, n = 18; OP, n = 20; LC, n = 19; OC, n = 11). RESULTS: Results are expressed as mean +/- SEM or median (interquartile range). Fasting plasma glucose levels were similar in the four groups but the plasma glucose area was higher after oral glucose (75 g) in both the lean and obese PCOS groups than in their controls (LC 32.4 +/- 0.7 vs LP 35.2 +/- 1.2, P < 0.01; OC 34.7 +/- 1.8 vs OP 37.8 +/- 1.5 mmol/l/3 h, P < 0.01). Insulin sensitivity was significantly reduced in obese PCOS women (LC 196 +/- 9 vs LP 179 +/- 9, NS; OC 168 +/- 12 vs OP 133 +/- 9 mmol/l/min, P < 0.01). Total serum cholesterol levels were similar in the four groups but HDL2-cholesterol was reduced in both obese and lean PCOS (LC 0.42 (0.38-0.62), LP 0.31 (0.26-0.44), P < 0.05; OC 0.34 (0.21-0.47), OP 0.21 (0.12-0.32) mmol/l, P < 0.01). Total HDL cholesterol was decreased significantly only in the obese PCOS group. Body mass index correlated significantly and negatively with total HDL-cholesterol and with HDL2-cholesterol levels both within the PCOS group and the control women. Using multiple regression insulin insensitivity contributes significantly beyond BMI to the low HDL-cholesterol in women with polycystic ovaries. CONCLUSION: Polycystic ovary syndrome is associated with biochemical risk factors for premature vascular disease, which cannot be explained by obesity alone. PMID- 8729523 TI - Circulating levels of growth hormone, insulin-like growth factor-I and growth hormone binding protein in normal women of advanced reproductive age. AB - OBJECTIVE: Women experience an age-related decline in fertility despite regular ovulatory cycles and normal production of ovarian steroids. Growth hormone and IGF-I are both reported to decline with age, and there is evidence that both hormones promote intraovarian actions of gonadotrophins. The purpose of this study was to characterize circulating levels of GH and IGF-I in normal, older reproductive age women with ovulatory cycles. DESIGN: Prospective, controlled. PATIENTS: Twenty-eight regularly cycling older (n = 16) and younger (n = 12) women were recruited for daily blood sampling throughout a menstrual cycle. MEASUREMENTS: Serum obtained from daily blood sampling was analysed for LH, FSH, oestradiol (E2) and progesterone (P). Serum obtained from frequent sampling during the admission was analysed for pulsatile GH secretion. IGF-I and GH binding protein (GHBP) were also measured in subsets of the two age groups. RESULTS: All subjects exhibited normal patterns of LH, FSH, E2 and P consistent with ovulatory cycles. There were no differences between the two age groups in integrated 24-hour GH secretion or in GH pulse amplitude or frequency. There were no differences in GH secretion between the early follicular and miduluteal phases when data were combined for the two subject groups. Plasma concentrations of IGF I were significantly lower throughout the cycle in the older women. There were no significant differences in levels of GHBP across the cycle or between the two age groups. CONCLUSIONS: IGF-I decreases with age in women without identifiable changes in the amount or pattern of GH secretion or in circulating GHBP concentrations. Decreased IGF-I production may be related to decreased ovarian gonadotrophin sensitivity in older reproductive age women. PMID- 8729524 TI - Medroxyprogesterone acetate enhances the spinal bone mineral density response to oestrogen in late post-menopausal women. AB - OBJECTIVES: The relative contributions of administered oestrogen and progestin to protection of the post-menopausal skeleton remain unclear. We have compared the effect on spinal bone mineral density of continuous combined oestrogen/medroxyprogesterone therapy with that of unopposed oestrogen in late post-menopausal women. DESIGN: Observational study. SUBJECTS: Seventy-three osteoporotic post-menopausal women attending the Auckland Hospital Bone Clinic. Twenty-three hysterectomized women were treated with unopposed oestrogen (0.625 mg oral conjugated oestrogens daily), and 50 non-hysterectomized women were treated with continuous combined oestrogen/medroxyprogesterone therapy (5 mg medroxyprogesterone acetate and 0.625 mg oral conjugated oestrogens daily). MEASUREMENTS: Baseline and one-year measurements of lumbar spine bone mineral density were performed using dual-energy X-ray absorptiometry. RESULTS: After one year of therapy, spinal bone mineral density increased by 6.6% (95% confidence interval 5.6-7.6, P < 0.001 vs baseline) in the combined oestrogen/medroxyprogesterone therapy group, compared to 4.0% (CI 2.4-5.7, P < 0.001 vs baseline) in the unopposed oestrogen group (difference between means 2.6% (CI 0.8-4.4, P < 0.01). CONCLUSION: In osteoporotic post-menopausal women, one year of continuous combined oestrogen/medroxyprogesterone therapy is associated with a 65% greater increment in spinal bone mineral density than is observed in response to unopposed oestrogen. The prescription of combined oestrogen/progestin therapy should be considered in osteoporotic post-menopausal women who have undergone hysterectomy, in order to maximize the skeletal protection provided by hormone replacement therapy. PMID- 8729525 TI - Medroxyprogesterone and bone mineral density response to oestrogen. PMID- 8729526 TI - Pituitary enlargement. PMID- 8729527 TI - High prolactin levels may be missed by immunoradiometric assay in patients with macroprolactinomas. AB - OBJECTIVE: Large amounts of antigen may produce falsely low values in immunoradiometric assays due to the so-called high dose, hook effect. The study was designed to identify the clinical and biochemical features of patients with pituitary macroadenomas in whom a high dose PRL hook effect was documented. DESIGN: The clinical and biochemical features of four patients with the high dose PRL hook effect were compared with those of 54 patients with pituitary non functioning adenomas and 11 with macroprolactinomas who underwent transsphenoidal microsurgery between 1989 and 1994. MEASUREMENTS: The presence of the high dose PRL hook effect was confirmed by an increase in the initial PRL concentration when the immunoradiometric assay was processed after dilutions of the serum. This phenomenon was observed in 5.8% (4/69) of patients with pituitary macroadenomas. Undiluted median (range) PRL levels were 9140 (1530-83850), 1530 (162-3210) and 2110 mU/l (1470-45,000) in patients with macroprolactinoma, non-functioning macroadenoma and the hook effect, respectively. In patients with the hook effect, the median PRL levels increased to 384,720 (317,520-950,000) mU/l when the assay was performed after serum dilution. The proportion of males was 9.9% (1/11) in the macroprolactinoma group, 46.3% (25/54) in the non-functioning macroadenoma group and 100% (4/4) in patients with the hook effect. Patients with prolactinoma and non-functioning adenoma had mean tumour sizes of 20 +/- 9 and 27 +/- 11 mm (SD), respectively, while in the hook effect group it was 51 +/- 10 mm. CONCLUSION: This study suggests that the high dose PRL hook effect is observed particularly in patients with very large tumours. The immunoradiometric PRL assay must be performed with serum dilution in order to overcome the high dose PRL hook effect in all new patients with pituitary macroadenomas who may have a prolactinoma. PMID- 8729528 TI - The prevalence of severe growth hormone deficiency in adults who received growth hormone replacement in childhood [see comment]. AB - OBJECTIVE: The few previous studies in which reassessment of GH status has been carried out in young adults treated with GH therapy for childhood GH deficiency concentrated on determining the incidence of 'transient GH deficiency'. As the benefits of GH replacement therapy in adults become increasingly appreciated, it is likely that GH therapy started in childhood for GH deficiency will be continued into adult life in many of those with severe GH deficiency. The aim of this study is to determine how many patients who received GH replacement therapy in childhood until completion of growth have GH deficiency severe enough to be considered for GH replacement therapy in adult life. DESIGN: Retrospective analysis of the peak GH responses to provocative stimuli performed at the time of diagnosis of GH deficiency in childhood and at the completion of growth after GH replacement therapy had been stopped. PATIENTS: Eighty-eight adults (49 male, 39 female) who had received GH therapy in childhood for a diagnosis of GH deficiency. The aetiology of the GH deficiency included craniopharyngioma, radiation induced associated with either a cerebral tumour or acute lymphoblastic leukaemia, histiocytosis-X and idiopathic. MEASUREMENTS: In childhood the original diagnosis of GH deficiency was based biochemically on the failure of the peak GH response to reach 20 mU/l during two provocative tests in 59 of the 88 patients and to a single test in the remaining 29. A total of 147 tests were performed, the most common being an insulin tolerance test (ITT, n = 72) and an arginine stimulation test (AST, n = 53). At reassessment in young adult life 146 tests were performed (74 ITT, 64 AST). Severe GH deficiency was defined arbitrarily as a peak GH response of less than 9 mU/l to a single (n = 33) or to two (n = 55) pharmacological stimuli. RESULTS: By definition all patients were considered GH deficient at the time of initial diagnosis. A peak GH response of less than 9 mU/l was seen in 64.8% at initial assessment and 60.2% at reassessment. Analysis in aetiological terms, however, showed that between assessments the incidence of severe GH deficiency increased in the group of radiation induced (48.8 vs. 55.8%) but decreased in the idiopathic group (78.1 vs. 53.1%). Out of the 55 patients who underwent two tests at reassessment, 47.3% of those with a GH peak less than 9 mU/l at one test had a GH peak greater than 9 mU/l at the second test. Fifteen of the 55 patients had additional pituitary hormone deficiencies and all 15 had a GH peak below 9 mU/l in both tests. CONCLUSIONS: Our study suggests that all children who have received GH replacement therapy in childhood should undergo reassessment of GH status in young adult life. Between 40 and 60% of such patients merit consideration for GH therapy in adult life depending on the definition of severe GH deficiency in use. Patients with isolated GH deficiency should undergo two provocative tests of GH secretion, but those with additional anterior pituitary hormone deficiencies require only one test at reassessment. PMID- 8729529 TI - Growth hormone replacement treatment in adult patients. PMID- 8729530 TI - Growth hormone treatment affects brain neurotransmitters and thyroxine [see comment]. AB - OBJECTIVE: Binding sites specific for growth hormone have been identified in the brain, but the action of GH on the central nervous system is still poorly understood. DESIGN: In a double-blind, placebo-controlled 21-month trial with a cross-over design, with each treatment period lasting for 9 months, we investigated the long-term effect of GH on the cerebrospinal fluid (CSF) concentrations of some brain neurotransmitters and thyroid hormones of importance for mood and cognition. PATIENTS: Twenty-four patients with documented GH deficiency acquired in adult life took part. RESULTS: Analysis of CSF collected at the end of the two treatment periods showed that the GH concentration was related to the administered dose of rhGH (r = 0.56, P = 0.0044). After rhGH treatment the concentration of the dopamine metabolite homovanillic acid (HVA) had decreased from 218 +/- 80 to 193 +/- 82 nmol/l (P = 0.002) and that of the excitatory acid aspartate had increased from 233 +/- 81 to 313 +/- 116 nmol/l (P = 0.032). No effects were observed on the concentrations of 5-hydroxyindoleacetic acid (the serotonin metabolite) and of 3-methoxy-4-hydroxyphenyl glycol (the noradrenaline metabolite), or on those of glutamate, glycine and beta-endorphin. However, both CSF and serum levels of free T4 decreased, from 19.8 +/- 6.1 to 16.6 +/- 5.7 nmol/l (P = 0.0002) and 17.0 +/- 5.0 to 13.7 +/- 4.3 nmol/l (P = 0.0001), respectively. The concentration of total T3 was not measurable in CSF but increased in serum from 1.41 to 1.53 nmol/l (P = 0.01). CONCLUSION: The study demonstrates a passage of GH from the circulation into the CSF. The observed changes in homovanillic acid and free T4 are similar to those reported after successful treatment of depressive disorders with antidepressant drugs, and may reflect a beneficial effect of GH on mood and behaviour. PMID- 8729531 TI - Growth hormone treatment, brain neurotransmitters and thyroxine. PMID- 8729532 TI - Growth hormone secretion in normal prepubertal children: importance of relations between endogenous secretion, pulsatility and body mass. AB - OBJECTIVE: Although a negative relation between GH secretion and body mass has been described, few studies have been carried out in prepubertal subjects of normal weight and height. The aim of the present study was to examine the relations between GH secretion, pulsatility and body mass throughout the wide range of weights in normal prepubertal children. METHODS: We determined integrated GH concentrations during the day (IC-GHD), night (IC-GHN), and 24 hours (IC-GH24) in 46 prepubertal children (26 males, 20 females) of normal height and growth velocity and with a Z-score (BMI-SD) between +2 and -2 SD. In addition, in 28 of these patients randomized by their BMI-SD, the secretory profile was studied by the Pulsar program: area under curve (AUC), peak number (NP), high spontaneous peak of secretory profile (HSP), and area of pulses (AP) which were considered large (L) or small (S) if their amplitude was greater or less than 4 micrograms/l (8 mU/l). RESULTS: BMI-SD correlated negatively with IC GH (IC-GHD, IC-GH24, r = -0.58, P < 0.001) and with Pulsar parameters r = -0.48, P < 0.001; IC-GHN, r = -0.45, P < 0.001; (AUC-24, r = -0.73, P < 0.0001; HSP, r = -0.65, P < 0.0001). The NP correlated negatively with BMI-SD (r = -0.51, P < 0.005) at the expense of NLP (r = -0.54, P < 0.003) without being significantly correlated with NSP. This inverse correlation between BMI-SD and NLP was greater during daytime (NLDP, r = -0.60, P < 0.001) and was not significant at night. NSP did not correlate with BMI-SD. A similar relation was observed with respect to pulse area (ALP, r = -0.38, P < 0.05; ALDP, r = -0.46, P < 0.02;) and was not significant for ALP during the night or for ASP in any period. CONCLUSIONS: We suggest that it should be necessary to relate GH secretion to body mass in prepubertal children before defining their secretion as normal or pathological. PMID- 8729533 TI - Hypergastrinaemia in Cushing's syndrome: pituitary origin or glucocorticoid induced? AB - OBJECTIVES: Several reports have shown an increase in serum gastrin levels in patients with Cushing's syndrome (CS). However, the actual origin of this hypergastrinaemia is not known. Two hypotheses have been proposed: concomitant ACTH and gastrin secretion by corticotrophic pituitary adenomas or hypergastrinaemia induced by hypercortisolism. DESIGN AND PATIENTS: We performed simultaneous, bilateral inferior petrosal sinus (IPS) sampling in nine patients with Cushing's disease (CD), proven by histological studies. In all of them, blood samples were taken from both IPS and a peripheral vein to measure plasma ACTH and serum gastrin. In addition, we measured peripheral serum gastrin levels after an overnight fast in 10 patients with CS (seven with pituitary tumours and three with adrenal tumours) before and after surgical treatment. RESULTS: Petrosal-peripheral and interpetrosal gradients of ACTH were higher than 2.0 and 1.4, respectively, confirming the pituitary origin of ACTH. Mean serum gastrin levels were 149.1 +/- 53.6 ng/l in peripheral vein, 183.4 +/- 71.7 ng/l in dominant IPS and 181.4 +/- 68.9 ng/l in non-dominant IPS. No significant differences in gastrin concentrations in these locations were found. Mean preoperative gastrin level in patients with CD was 194.6 +/- 47.9 ng/l, whereas in patients with adrenal tumours it was 247.3 +/- 125.9 ng/l. After surgical treatment, the gastrin levels decreased to 62.1 +/- 13.2 ng/l (P < 0.05) and 90.3 +/- 50.3 ng/l (NS), respectively. CONCLUSIONS: These results suggest that hypergastrinaemia is a common finding in patients with Cushing's syndrome. The lack of significant petrosal-peripheral gradient in individuals with Cushing's disease and the reduction in gastrin level following adrenal tumour resection argues against the hypothesis of a predominantly pituitary source of gastrin, suggesting a glucocorticoid related mechanism as an explanation for the hypergastrinaemia. PMID- 8729535 TI - Endoscopic adrenalectomy: the adrenals under the scope? PMID- 8729534 TI - Structure of the thyrotrophin-releasing hormone receptor in human pituitary adenomas. AB - BACKGROUND AND OBJECTIVE: TRH acts on specific G-protein coupled receptors sited in cells of the anterior pituitary gland. Pituitary tumours expressing either TSH, PRL or GH may respond to TRH by enhanced, blunted or paradoxical hormone release. Non-functioning pituitary tumours may also show abnormal responses to TRH. Little is understood of the mechanisms regulating inappropriate hormone release in these tumours. Activating or inactivating mutations found in G-protein coupled receptors have been implicated in human pathological conditions. Mutations in the G-protein coupled TRH receptor might be involved in the aetiology of pituitary adenomas resulting in aberrant hormone secretion. We therefore screened samples of pituitary adenomas for the presence of somatic mutations in the TRH receptor gene. PATIENTS: Pituitary adenoma tissue samples were obtained at surgery from 50 patients with pituitary adenoma (17 acromegaly, 15 prolactinoma, 11 TSH-secreting and 7 non-functioning adenoma) along with blood samples to provide lymphocyte DNA as control sequence. METHODS: Genomic DNA was extracted from adenoma and lymphocyte samples and the entire coding region of the TRH receptor was amplified using 5 overlapping pairs of PCR primers. The PCR products were analysed for mutations by non-denaturing polyacrylamide gel electrophoresis which reveals single-strand conformational polymorphisms (SSCP) as a mobility shift in product migration. Wild-type and mutant TRH receptor cDNA were similarly analysed to confirm the sensitivity of the method. Additionally, PCR products were ligated into a PCR cloning vector and DNA sequencing carried out to confirm the findings of SSCP analysis. RESULTS: The human TRH receptor retained normal wild-type sequence in the large group of TSH secreting, PRL secreting, GH secreting and non-functioning pituitary adenomas investigated in this study. CONCLUSION: Our observations suggest that the TRH receptor structure is normal in TSH secreting, PRL secreting, GH secreting and non-functioning pituitary adenomas. It is therefore unlikely that the TRH receptor is involved in the pathology associated with the types of pituitary adenomas investigated in this study. It is possible that some other component of the pathway controlling TRH-signalling events may be implicated in pituitary tumorigenesis. PMID- 8729536 TI - Apparent mineralocorticoid excess due to 11 beta-hydroxysteroid dehydrogenase deficiency: a possible cause of intrauterine growth retardation. AB - A Japanese boy with apparent mineralocorticoid excess (AME) is described. He was born with intrauterine growth retardation (IUGR) and elevated serum level of creatine phosphokinase (CPK). He was studied at 2 years of age because of polyurea and polydipsia of one year's duration and was found to have hypokalaemic alkalosis and sustained hypertension. His plasma renin activity and aldosterone levels were always low and his ratio of urinary tetrahydrocortisol plus allo tetrahydrocortisol to that of tetrahydrocortisone was very high. Therefore, AME due to 11 beta-hydroxysteroid dehydrogenase (11 beta-HSD) deficiency was diagnosed. He was successfully treated with a combination of spironolactone and nifedipine for at least 16 months. His blood pressure, plasma pH and serum potassium levels were normalized by this treatment, but serum CPK level remained high. We researched the birth records of previously reported AME cases and found that IUGR is a characteristic feature of AME. The mechanism by which IUGR occurs in AME is discussed and we speculate that 11 beta-HSD might be deficient in the placenta and/or fetal tissues, as well as in the kidney, in AME. An explanation for the elevated CPK could not be found. PMID- 8729537 TI - Presidents who have caused a stir. First general meeting; Monday, 11th October 1993. PMID- 8729538 TI - The accident & emergency department and its interface with general practice. Ordinary meeting; Monday, 25th October, 1993. PMID- 8729539 TI - In the public interest. Ordinary meeting; Monday, 22nd November 1993. PMID- 8729540 TI - Consent to medical treatment: the law and ethics. Ordinary meeting; Monday, 24th January, 1994. PMID- 8729541 TI - Medical eponyms updated. Ordinary meeting; Monday, 14th February 1994. PMID- 8729542 TI - Language, poetry and attitude to sound. Ordinary meeting; Monday, 28th February, 1994. PMID- 8729544 TI - The inner narrative of evil. Ordinary meeting; Monday, 28th March 1994. PMID- 8729543 TI - The influence of eye disorders on the artist. Lettsomian lecture Monday, 14th March 1994. PMID- 8729545 TI - The ruin of the constitution: the early interpretation of gout. Ordinary meeting Monday, 25th April 1994. PMID- 8729546 TI - The annual oration--Future patterns of medical practice. Second general meeting; Monday, 9th May 1994. PMID- 8729547 TI - Sir John McNee bequest summary of a report on a period of elective study immunization rates in Moscow preschool children. AB - We describe childhood immunization rates in a sample of Muscovite children and attitudes towards child immunization. Of 19 children, 57% of children were up-to date with diphtheria and tetanus immunization, 47% with pertussis, 59% with oral poliovirus, 91% with tuberculosis and 51% with measles. There was no evidence of fear about infections by needles, preference for alternative medicine or unavailability of vaccines. Immunization rates amongst pre-school Muscovite children are low but not that much different from those in some areas of the United Kingdom in the 1980s or areas of the United States at present. Medical advice against immunization was the main reason for failure. PMID- 8729548 TI - Pharmacokinetics. PMID- 8729549 TI - Ambulatory blood pressure monitoring in hypertension. PMID- 8729550 TI - Successful management of hyperemesis gravidarum using steroid therapy. AB - Hyperemesis gravidarum causes uncontrollable vomiting, severe dehydration and muscle wasting in pregnancy and usually requires weeks or months of intravenous fluid therapy. A consecutive series of 7 women with hyperemesis gravidarum were treated with high-dose steroid therapy. Vomiting stopped within 3 h of the first dose of intravenous hydrocortisone in all patients. Maintainence oral prednisolone therapy in doses up to 45 mg/day permitted discharge from hospital within days, resumption of normal eating, reversal of muscle wasting and regain of lost weight (mean loss from prepregnant weight 10.5 +/- 4.3 kg). Prednisolone in doses of 15 mg/day or more was required for 10.6 +/- 4.7 (range 6-20) weeks. High-dose prednisolone therapy is effective in suppressing symptoms of intractable hyperemesis gravidarum and allowing normal maternal nutrition. PMID- 8729551 TI - Clinical measurement of swallowing in health and in neurogenic dysphagia. AB - We studied clinical features potentially related to dysphagia and three indices from a timed test of swallowing--average volume per swallow (ml), average time (s) per swallow and swallowing capacity (ml/s)--in 181 screened healthy adults and 30 patients with motor neurone disease (MND). In healthy adults, age, sex and height accounted for 44.3% and 55.6% of the variance of log average volume per swallow and log swallowing capacity, respectively. Symptoms and signs were more prevalent in the MND group and were associated with reduced swallowing capacity and reduced average volume per swallow; repeatability studies on these two indices in both groups showed that the median difference between the mean of two recordings on successive days and the mean of all recordings (6-15 over 3 days) was < 5% (maximum third quartile 12.8%, indices expressed as percent predicted according to age and sex). Using this simple bedside test, swallowing function can be quantified on a ratio scale and expressed as percent of that predicted by age and sex; such information may improve the predictive value of clinical assessment and provides a practical way of monitoring change in patients with dysphagia. PMID- 8729552 TI - Chronic hepatitis C and B-cell non-Hodgkin's lymphoma. AB - Hepatitis C virus (HCV), a hepatotropic and lymphotropic virus, is the major causative agent of nonA-nonB chronic hepatitis; moreover, it is frequently associated with benign and malignant lymphoproliferative disorders such as mixed cryoglobulinaemia and B-cell non-Hodgkin's lymphoma (NHL). We investigated the clinical and virological features of B-cell NHL complicating chronic hepatitis C in a series of 10 patients (M/F 1/9; mean age 63 +/- 6SD years). The malignancy appeared after median 8 +/- 4SD years from onset and was low-grade in six patients, intermediate in three, and high-grade in one. 'One-tube nested' PCR detected serum HCV RNA and viral ongoing replication in both fresh and cultured peripheral lymphocytes in all ten. Analysis of HCV genotypes showed a relatively higher prevalence of 2a/III type compared with unselected chronic hepatitis C (50% vs. 15%). In one patient, HCV RNA was also found in the neoplastic bone marrow and lymph-node specimens. B-cell NHL can complicate chronic hepatitis C and affect the overall prognosis of the disease. The increasing frequency of chronic hepatitis C worldwide suggests that the actual prevalence of this complication may be underestimated. Careful clinical work-up at diagnosis and during follow-up is particularly recommendable. PMID- 8729553 TI - Classifying extraintestinal non-typhoid Salmonella infections. AB - Non-typhoid Salmonella infection in man has been divided into five clinical groups: gastroenteritis, enteric fever, bacteraemia, chronic carrier state and localized infection. This classification has neither pathogenic nor prognostic significance. We retrospectively reviewed the charts of 183 patients with extraintestinal salmonellosis who presented to our institution during a period of 32 years. Patients were classified into four groups: primary bacteraemia (PB), enteritis-associated bacteraemia (secondary bacteraemia) (SB), digestive focal infection (DI) and non-digestive focal infection (NDI). Sex, age, acquisition, underlying disease and outcome were compared between patients with bacteraemia and diseases with focal infection. The differences found between PB and SB were: community acquisition (66% in PB and 85% in SB, p = 0.06) severe immunosuppression (53% in PB and 15% in SB, p < 0.001) and mortality (37% in PB and 3% in SB, p < 0.001). The differences found between NDI and DI were: age over 60 years (45% in NDI and 18% in DI, p < 0.05), severe immunosuppression (51% in NDI and 12% DI, p < 0.001) and associated bacteraemia (38% in NDI and 6% in DI, p < 0.001). This classification of extraintestinal salmonellosis may have pathogenic and prognostic implications, and could help us to understand the clinical significance of this disease. PMID- 8729554 TI - Factors determining the maintenance dose of warfarin in Chinese patients. AB - Chinese patients are reportedly more sensitive than Caucasians to the anticoagulant effect of warfarin. We examined warfarin dose requirements and their determinants in 151 Chinese out-patients on stable maintenance dose of warfarin with international normalized ratio of 2 to 2.5. Mean daily warfarin requirement was 3.3 +/- 1.4 mg, much lower than reported doses in Caucasian patients. The most important determinant was age (r = -0.43, p < 0.001), with progressively lower warfarin requirement with increasing age (p = 0.0001). There was a weaker association with body weight (r = 0.20, p = 0.01). Patients with chronic rheumatic heart disease tended to require a smaller dose than those with heart valve replacements (2.94 +/- 1.24 vs. 3.69 +/- 1.42 mg, p < 0.01). We confirm that Chinese patients require a smaller dose of warfarin for the same degree of anticoagulation. Age is the most important factor affecting dose requirement, although body weight and underlying disease also play a role. PMID- 8729555 TI - Biochemical markers of alcohol abuse. AB - Excessive alcohol consumption is a major health problem in the UK leading to both serious morbidity and mortality. This study compared newer potential biochemical markers of excessive alcohol consumption [carbohydrate-deficient transferrin (CDT), mitochondrial AST (mAST) and alpha glutathione-s-transferase (alpha-GST)] with conventional markers (AST, ALT, GGT, MCV). Patients (n = 85) were enrolled in the study and subdivided into several groups on the basis of alcohol consumption. Patients with non-alcoholic liver disease (NALD) (n = 40) were also enrolled. All the markers, with the exception of the ratio mAST/total AST were significantly higher in heavy drinkers/alcoholics compared to teetotallers/social drinkers (p < 0.05). mAST and AST/ALT ratio were significantly higher in alcoholics compared to NALD (p < 0.01), whereas ALT was higher in the NALD group (p < 0.05). Multivariate discriminant function analysis (Wilks method) demonstrated that the logarithmic functions of AST/ALT ratio and mAST could correctly classify 87.9% of cases into either the alcoholic or NALD groups. ROC plot analysis showed that AST, mAST and GGT were the best markers at distinguishing heavy consumption of alcohol from lesser levels and that AST/ALT ratio and mAST were the best in distinguishing alcoholics from NALD. In conclusion, none of the newer biochemical markers, with the exception of mAST, offers any major advantage over the conventional markers. PMID- 8729556 TI - Angiotensin-converting-enzyme inhibitors in the management of cardiac failure: are we ignoring the evidence? AB - The benefits of angiotensin-converting enzyme (ACE) inhibition in the management of cardiac failure have been extensively documented. However, little is known about its impact upon the investigation and management of this condition. We assessed how patients diagnosed as having cardiac failure were investigated, which patients were treated with ACE inhibitors and with what dosages. We reviewed the case notes of all 343 patients discharged from Aberdeen Royal Infirmary 1 July-31 December 1992 with a diagnosis of cardiac failure. In addition, a questionnaire was sent to the general practitioners of the 166 patients still alive in October 1994. Only 40% of patients were discharged from hospital on ACE inhibitors. In 58.8%, the diagnosis of cardiac failure was based purely on clinical or radiological grounds. At discharge, 76.1% of patients were on lower doses of ACE inhibitors than those used in the major survival studies; with 68.9% receiving similar doses two years later. The majority of patients with heart failure are under-investigated and under-treated. PMID- 8729557 TI - Geographical variation in the referral of patients with chronic end stage renal failure for renal replacement therapy. AB - The number of dialysis units per million population is low in the UK by comparison with other European countries, and this may affect the referral of patients for renal replacement therapy. We used a Poisson regression model to analyse the spatial distribution of all 539 adult patients resident in south-west Wales (Dyfed and West Glamorgan) who started chronic renal replacement therapy between April 1985 and March 1994. Controlling for patient age, population distribution, socio-economic variables and ethnic group, there was a significant negative relationship between referral rates and distance of residence from the renal unit for patients aged over 60 years, but not for younger patients. The prevalence of renal replacement therapy rose from 128 to 454 per million in Dyfed, and from 188 to 647 per million in West Glamorgan, between 1985 and 1995. PMID- 8729558 TI - Viral hepatitis' G string. PMID- 8729559 TI - Hypertension in porphyria--an understated problem. PMID- 8729560 TI - Brainstem hypoperfusion in CFS. PMID- 8729561 TI - Dyslexia: the link with visual deficits. AB - Some research reports suggest that visual anomalies may have a causative role in dyslexia, and on this basis certain forms of therapy have been proposed. Recently, we have published the initial results of a matched group study which found dyslexia to be associated with binocular instability, reduced amplitude of accommodation, and reduced contrast sensitivity for both low spatial frequencies and uniform field flicker. The binocular instability was best identified by measuring the vergence amplitude: a modified Dunlop test failed to differentiate reliably between the two groups. Here, we report a significant correlation between flicker threshold and binocular instability, thus linking sensory and motor visual correlates of dyslexia. We also present further new analyses on the interaction between optometric variables and the psychometric measurement of coding skills. The results of these analyses disagree with a recent claim that binocular vision anomalies might cause poor performance at coding tasks. Our studies indicate that visual characteristics are not the major aetiological factors in specific reading difficulty. PMID- 8729562 TI - Prevalence of spectacle and contact lens wear in the UK fire service. AB - Despite the fact that firefighting has been shown to be a visually demanding occupation, the present visual standards for firefighters prohibit the use of spectacles or contact lenses operationally. To determine the proportion of the UK fire service currently using spectacles and contact lenses, and how many may benefit from optical correction, a questionnaire was distributed. This contained 30 questions and was sent to 1550 firefighters. The data from 1064 returned questionnaires have been analysed. The results show that approximately 20% of those serving in the fire service have used an optical correction at some time in their lives. When not working, approximately 6% of firefighters wear spectacles for distance vision and 15% wear them for reading. Only 0.4% of firefighters use contact lenses. Half of this contact lens wearing group use their lenses while on operational duty. In total, approximately 2% of firefighters use an optical correction for operational duties. The results indicate that a high visual standard at the time of entry into an occupation cannot guarantee good vision throughout an individual's career. In addition, an estimated 11,000 firefighters in the UK, who have spectacles or contact lenses, do not use them for operational duties. The present visual standards prevent this group of firefighters from obtaining their optimal visual performance at work. PMID- 8729563 TI - Elimination and minimization of the spherical aberration of intraocular lenses in high myopia. AB - By applying Seidel aberration theory we have studied the spherical aberration of intraocular lenses in place (SIL), and the total spherical aberration of pseudophakic eyes (S(IT)). The optimum lens shapes for zero SIL and S(IT) or for minimum SIL and S(IT) of 12 schematic, highly myopic eyes were investigated. When the power required for emmetropization is positive, the total Seidel spherical aberration of the whole eye cannot be zero but can only be minimized. If the power of the intraocular lens is negative, the total Seidel spherical aberration of the whole eye can be made zero, and one of the optimum lens shapes would be meniscus with the convex surface towards the retina. In addition to improving the quality of the image, the lens has advantages from the physiological point of view, since it helps to maintain the structure of the eye. PMID- 8729565 TI - Contrast and luminance as parameters defining the output of the VERIS topographical ERG. AB - The Visual Evoked Response Imaging System (VERIS I) topographical electroretinogram system (EDI Associates, San Francisco, CA) allows measurement of the response of large numbers of retinal areas simultaneously. This paper examines ERG responses derived with the VERIS system to changes in target contrast and to local reductions in luminance. There is a linear reduction of response with reductions of target contrast. Neutral density filters were placed over part of the display, to mimic the localised reductions in response which occur in glaucoma, age-related maculopathy or diabetic retinopathy. There is a definite reduction in response seen with a 0.4 ND filter, indicating that the system should have similar sensitivity in detection of retinal lesions to the commonly used visual field analysis systems. PMID- 8729564 TI - Proximal contribution to a linear static model of accommodation and vergence. AB - To determine the influence of target proximity on accommodation and vergence under both open- (OL) and closed-loop (CL) viewing conditions, a static interactive feedback model, which included proximal accommodation (PA) and proximal vergence (PV) inputs, was developed and analysed quantitatively. It was based on an earlier static dual-interactive feedback model of accommodation and vergence. The proximal inputs were added to both the accommodative and vergence loops at the output of the respective controllers. The values of the PA and PV gains were obtained from experimental dual-OL data. The model equations were analysed over a stimulus range of 1 to 6 D (or MA). It was found that under the dual-OL condition, the contribution of PA to the overall accommodative output ranged from 42.5 to 81.6%, whereas the contribution of PV to the overall vergence output ranged from 56.1 to 88.5%. In contrast, under all other stimulus conditions, with the exception of the PA contribution to the accommodative output under the Aol, Vcl condition (26.5-41.0%), the relative contributions were much smaller, ranging from 0.04 to approximately 7.0%. In particular, under the dual CL condition, representing the normal binocular visual feedback condition, the relative contributions were only 4.0 and 0.04% for PA and PV, respectively. Thus, although the relative contributions of PA and PV were large under the dual-OL condition, they were generally very small under the various CL conditions that simulated more naturalistic viewing situations. Nevertheless, proximity may still play an important role by providing cues for attaining coordinated and harmonious motor responses under specific viewing conditions. PMID- 8729566 TI - Relative roles of resolution and spatial interference in foveal and peripheral vision. AB - The rate of change of visual performance with increasing eccentricity of fixation is known to be task dependent. In order to examine eccentricity-related changes in visual performance when a combination of two different factors influence thresholds, visual acuity in the presence of flanking elements was investigated across the visual field. Interference zones increased in extent with eccentricity at a much faster rate than did resolution thresholds. E2 values, quantifying the eccentricity-related magnification required, were 1-2 degrees for resolution and 0.1-0.2 degrees for spatial interference. E2 values for resolution in the presence of flanks depended on the proximity of the flanks and their 'weighting' in the task. From the results, the magnitude and extent of spatial interference across the visual field can be described quantitatively. Further, the results serve as an example of how observed variations in peripheral threshold gradients might be achieved as a combination of underlying factors with different E2 values. PMID- 8729567 TI - Spectral sensitivity as revealed by isolated step onsets and step offsets. AB - Opponent-colour spectral sensitivity functions are traditionally elicited with increments; the temporal presentation of these increments is usually step onset and step offset. However, there are both physiological and psychophysical data suggesting differences in sensitivity for spectral step onsets and step offsets. The purpose of this study was to elicit opponent-colour spectral sensitivity functions with isolated step onsets and offsets, and to compare the form and sensitivity of these functions. Spectral sensitivity functions were determined for large, long duration, step onset and step offset stimuli. Four human observers participated in the study. The opponent-colour functions obtained for onsets and offsets are largely similar in both form and sensitivity. Of particular interest is the finding that there is no reduction in sensitivity for blue offsets. These data are consistent with recently published psychological data and apparently at variance with physiological data that suggest a paucity of S-cone OFF-neurones in the primate retina and dorsal lateral geniculate nucleus (dLGN). PMID- 8729568 TI - Development of pupillary responses to grating stimuli. AB - Pupillary responses to stationary, achromatic sine wave gratings were measured in adults and in 1- and 3-month-old infants. Measurements of behaviourally determined contrast sensitivity were also obtained from the infant subjects. Contrast sensitivity functions were typical of those previously described: band pass in nature with sensitivity increasing markedly with age. The pupil grating response amplitudes also exhibited a spatial frequency and age dependence similar to that observed for contrast sensitivity. These findings provide further evidence that the neural mechanisms which underlie visual sensitivity to contrast also mediate pupillary responses to spatial structure. PMID- 8729569 TI - Differences in apparent contrast in yellow and white light. AB - The purpose of this study was to investigate suprathreshold contrast perception in white and yellow light. The contrast of black-on-white letters was adjusted to match the perceived contrast of yellow-on-yellow letters presented simultaneously on a video display. At lower contrasts (7-15%), the apparent contrast of yellow letters was slightly enhanced compared to black-on-white letters (mean enhancement = 23%), but this effect diminished with increasing contrast. The slight enhancement in yellow light was independent of letter size, and could not be explained by luminance differences between yellow and white displays. This effect may relate to the subjective improvement often reported when wearing yellow (blue-blocking) lenses. PMID- 8729570 TI - The source of disparity vergence innervation determines prism adaptation. AB - A physiological model of prism adaptation argues that adaptation magnitude and rate are functions of the magnitude of reflex vergence innervation. It has also been shown that there is reduced prism adaptation magnitude for a given disparity stimulus where only peripheral sensory fusion is present (the 'eccentricity effect'). This study attempted to determine whether the eccentricity effect is attributable to reduced reflex vergence innervation in peripheral fusion. Experiments were run in a Maxwellian view haploscope. Convergence and divergence adaptation to 6 delta disparity stimuli were quantified for seven binocularly normal subjects using subjective heterophoria measurements. Vergence response during central or peripheral fusion was quantified objectively by infrared oculography. Six of seven subjects revealed an eccentricity effect. However, in three of seven subjects the eccentricity effect was not predictable from the manifest vergence response. The results suggest that the source of reflex vergence innervation affects prism adaptation. A model is proposed whereby different sources of reflex disparity vergence innervation stimulate prism adaptation by way of separate neurological pathways of differing responsiveness. PMID- 8729571 TI - Multi-meridional keratometry. AB - During conventional keratometry, the two principal meridia are identified and measurements of the anterior corneal curvature determined along these orientations. However, in cases of corneal irregularity, the principal meridia may be difficult to identify. Accordingly, the present study sought to evaluate two alternative procedures which do not require identification of the principal meridia. Instead, measurements of anterior corneal curvature were determined along either three or four preselected meridia and subsequently converted into the standard format for measurements of anterior corneal curvature. The study was performed on 25 visually normal subjects and the results compared with conventional two meridian keratometry findings. No significant differences were observed between the mean findings for the three and four meridian techniques. Furthermore, both procedures gave comparable results to the conventional two meridian method. Accordingly, multi-meridional keratometry may be valuable in cases of irregular astigmatism, for example produced by keratoconus or scarred corneas, since measurements can be obtained without the necessity for identification of the principal meridia. PMID- 8729572 TI - The proportion of confirmed glaucomas who have a family history of the disease. AB - In a large survey of glaucoma detection in practice, only 46 (10%) of 465 confirmed glaucomas had a family history of the disease. Together with other studies, this indicates that family screening would fail to detect the large majority of glaucoma cases. PMID- 8729573 TI - An appreciation. A. G. Bennett (1912-1994). PMID- 8729574 TI - A myocybernetic model of the lower urinary tract. AB - A biomechanical model of the lower urinary tract which is able to respond to input signals from a neural network is presented. The neural input is the starting point in the description of the relationships between the various physical parameters in the mechanical model of the bladder and the urethra. The cybernetics of the lower urinary tract are described on the basis of the muscle dynamics of simple models of both the detrusor in the bladder wall and the urethral sphincter. The urethral sphincter is not described as a variable resistance, like in other biomechanical models of the lower urinary tract, but is described on the basis of striated muscle dynamics. The forces produced by the detrusor and the urethral sphincter give rise to the bladder pressure and the urethral pressure. Using quasi-steady assumptions, the flow rate of urine is calculated as a result of the pressure difference between the bladder and the urethra. Parameters like the bladder volume, the flow rate and the pressure in the bladder can be compared with clinical data of urodynamic measurements. Simulation results show that the model is able to mimic both a filling and an emptying behaviour which resembles the behaviour of the lower urinary tract. By increasing the resistance of the urethra, a behaviour model of the lower urinary tract appears which is comparable with the pathology of urethral obstruction. A sensitivity analysis of various parameters in the model leads to a better understanding of the biomechanics of the lower urinary tract. PMID- 8729575 TI - The consequences of phenotypic plasticity in cyclically varying environments: a genetic algorithm study. AB - By "phenotypic plasticity" we refer to the capacity of a genotype to exhibit different phenotypes, whether in the same or in different environments. We have previously demonstrated that phenotypic plasticity can improve the degree of adaptation achieved via natural selection (Behera & Nanjundiah, 1995). That result was obtained from a genetic algorithm model of haploid genotypes (idealized as one-dimensional strings of genes) evolving in a fixed environment. Here, the dynamics of evolution is examined under conditions of a cyclically varying environment. We find that the rate of evolution, as well as the extent of adaptation (as measured by mean population fitness) is lowered because of environmental cycling. The decrease in adaptation caused by a varying environment can, however, be partly or wholly compensated by an increase in the degree of plasticity that a genotype is capable of. Also, the reduction of population fitness caused by a variable environment can be partially offset by decreasing the total number of genetic loci. We conjecture that an increase in genome size may have been among the factors responsible for the evolution of phenotypic plasticity. PMID- 8729576 TI - Evolution of transcriptional regulation system through promiscuous coupling of regulatory proteins with operons; suggestion from protein sequence similarities in Escherichia coli. AB - As an advanced molecular study of the problems of the evolution of organisms, the transcriptional regulation system is studied by investigating the amino acid sequence similarities between the proteins in the regulation system of Escherichia coli in which the data of sequenced proteins as well as of regulator regulon relationships are accumulated. The similarities between the proteins are calculated by the FASTA algorithm and their homology is also evaluated in terms of statistical significance with the use of the RDF2 program. This investigation reveals that the similarity between the regulatory protein and the regulated protein is hardly found, but many similarities are found between regulatory proteins and between regulated proteins. These similarity relations are compared with the regulator-regulon relationships ascertained experimentally. From this comparison, it is found that similar regulatory proteins rarely regulate the transcription of similar protein genes. As most of the highly similar proteins are considered to have diverged from a common ancestral protein, this finding strongly suggests the possibility that descendant regulatory proteins have been promiscuously coupled with descendant operons, independently of their ancestral regulator-regulon relationship, and that some of the couplings have been fixed by selection to form the present system of transcriptional regulation. The compatibility of such promiscuous coupling with regulatory organization is illustrated in the carbohydrate transport systems and the succeeding metabolic pathways, whose organization is comprehensive in sending nutritious substances to the central path of glycolysis under different environmental conditions. The benefit of flexibility in regulator-regulon relationships in evolutionary processes is also discussed in connection with the punctuational divergence of species in macroevolution and the cell differentiation in multicellular organisms. PMID- 8729577 TI - Study of correlations in DNA sequences. AB - We present a method for unified statistical analysis of short and long range correlations between various nucleotides in genomic DNA strands. The approach is based on the mutual study of Fourier structure factor spectra and pair correlation functions. The analysis of cross correlations in the different ranges of structural spectra permits identification of the main sources of correlations, namely, the coherent point mutations, coincident periodicities or large scale density variations. The technique for assessment of structural coupling between various genes in the genome is also described. PMID- 8729578 TI - Conscious mind as a field. PMID- 8729579 TI - Pharmacological stress electrocardiography--when is it indicated? PMID- 8729580 TI - Pharmacological stress echocardiography--practical considerations. PMID- 8729581 TI - Nuclear cardiology studies--principles for stress modality selection. PMID- 8729582 TI - Review of rationale for diagnostic modalities in combination. PMID- 8729583 TI - Lipid management: past, present, and future. AB - The therapeutic compounds available to the clinician treating lipid disorders in the 1990s enable the effective management of many patients. The development of the HMG-CoA reductase inhibitors is a major advance, and for the first time benefit in terms of overall mortality has been demonstrated in the impressive 4S study. What is needed now is effective implementation of screening and treatment of high-risk individuals. The UK continues to lag behind in this area. LDL cholesterol is the major target for therapy, but it is likely that over the next few years other lipid and lipoprotein parameters (HDL cholesterol, remnant particles, small dense LDL etc) will become more generally accepted targets for specific therapeutic intervention. Research continues to develop novel compounds targeting new regulatory steps in lipoprotein metabolism. In addition much progress is likely in targeting the arterial wall and atherogenesis other than specifically modulating plasma lipids. In the meantime the task of the clinician is to put into practice what is now known particularly about the secondary prevention of CHD to ensure that patients benefit from recent advances in knowledge. PMID- 8729584 TI - Development and pharmacology of fluvastatin. AB - Fluvastatin is the first synthetic 3-hydroxy-3-methylglutaryl coenzyme A (HMGCoA) reductase inhibitor to be approved for clinical use, and has been studied extensively in humans since 1986. It is structurally distinct from the other currently available HMGCoA reductase inhibitors (lovastatin, simvastatin, and pravastatin), leading to unique biopharmaceutical properties relative to the other agents of this class. Absorption of fluvastatin is virtually complete across all species, including man, and is not affected by the presence of food. Systemic exposure is limited, as fluvastatin is subject to first-pass metabolism, and the plasma half-life of the drug is approximately 30 minutes. Some 95% of a single dosage of fluvastatin is excreted via the biliary route, with less than 2% of this being the parent compound. Additionally, there is no evidence of circulating active metabolites or accumulation during chronic dosing. Studies of the effect of food on the pharmacokinetics of fluvastatin have demonstrated marked reductions in the rate of bioavailability--from 40% to 60%; however, a comparison of fluvastatin administration with the evening meal or at bedtime has revealed no significant differences in the extent of bioavailability (area under the curve) of these two regimens. Furthermore, no significant difference in pharmacodynamic effect (reduction in low-density lipoprotein cholesterol levels) could be ascertained between mealtime dosing and bedtime dosing. The pharmacokinetics of fluvastatin have also been assessed in various demographic groups. Relative to the general population, plasma concentrations of fluvastatin do not vary as a function of either age or gender. In addition, administration to a patient population with hepatic insufficiency resulted in a 2.5-fold increase in both the rate and extent of bioavailability relative to controls. Also, although minimal alterations of fluvastatin clearance in patients with renal insufficiency are anticipated due to limited renal excretion (5%), a study in this patient group is currently underway to examine this further. Interaction studies have been performed with fluvastatin and several drugs with which it might be coadministered. Cholestyramine, an anionic-binding resin, has a considerable effect in lowering the rate and extent of fluvastatin bioavailability. Although this effect was noted even when cholestyramine was given 4 hours prior to fluvastatin, this regimen did not result in diminished efficacy. Further, no effects on either warfarin levels or prothrombin times were observed in a study involving concomitant administration of warfarin and fluvastatin. Moreover, additional interaction studies with niacin and propranolol have not demonstrated any effect on fluvastatin plasma levels, and administration to a patient population chronically receiving digoxin resulted in no difference in the extent of bioavailability of digoxin relative to control data. The results generated to date in clinical pharmacokinetic studies with fluvastatin thus support its use in a broad population of hypercholesterolaemic patients. PMID- 8729585 TI - Treatment of dyslipidaemias with fluvastatin. AB - In many patients with primary hyperlipidaemia the statins have become the drugs of choice when diet alone has failed. The usual dosages of statins prescribed in most countries appear to be aimed at producing a reduction in levels of low density lipoprotein cholesterol (LDL-C) of about 25%. Fluvastatin is the newest member of this class of agent and the first completely synthetic compound. In dose-finding studies, fluvastatin has produced statistically significant (P < 0.001) reductions in LDL-C levels when used in dosages of 20 mg (21%) and 40 mg (27.4%), thus placing it within the range generally expected with usual dosages of other statins; additional studies have shown that the efficacy of fluvastatin is unaffected by gender and age. Moreover, fluvastatin produces the maximum reduction in LDL-C levels within 6 weeks and maintains this efficacy during long term treatment (LDL-C levels have been reported to fall by 27.4% over 2 years with fluvastatin 40 mg/day). In high-risk patients the LDL-C-lowering effect of fluvastatin 40 mg/day is maintained when compared with low-risk patients (26.6% vs 24.8%). A similar pattern has been observed in hypertensive patients, irrespective of the antihypertensive agent used to control blood pressure. Fluvastatin has been found to produce better LDL-C-lowering effects than bezafibrate. In addition to lowering LDL-C levels by the required amount, 20 and 40 mg (the equivalent of pravastatin 20 mg) dosages of fluvastatin significantly reduce levels of total cholesterol and triglycerides and increase high-density lipoprotein cholesterol levels. PMID- 8729586 TI - Safety profile of fluvastatin. AB - Statins are regarded as a well-tolerated class of drugs, particularly when compared with some of the older lipid-modifying agents, which have poor rates of compliance. Despite some early concern, the incidence of lens opacities observed in clinical studies involving statin use is no different from that in a normal ageing population. Similarly, the occurrence of insomnia with lipophilic agents appears to have been overemphasised and is not a clinically significant problem, irrespective of the statin under study. Fluvastatin is the newest representative of this class of agents; it has already been evaluated in thousands of patients who have hyperlipidaemia with and without additional risk factors. In controlled clinical studies, the incidence of the majority of adverse events observed with fluvastatin therapy is no higher than that seen with placebo, with the exception of gastrointestinal disturbances (known to be common to all stains). Nonetheless, the incidence of these effects seen with fluvastatin treatment is noted to be lower than that associated with cholestyramine or fibrate use. Elevations in levels of liver transaminases (aspartate aminotransferase and alanine aminotransferase) have been reported with fluvastatin therapy but have led to discontinuation of treatment with the same frequency as with placebo. Elevations in creatine kinase levels as a cause of discontinuing fluvastatin are not more frequent than with placebo. Drug-related myopathy and rhabdomyolysis have not been reported with fluvastatin therapy, and myalgia does not occur more frequently than with placebo. In terms of drug interactions, fluvastatin does not interfere with the efficacy of antihypertensive agents. In controlled clinical trials, the overall reported discontinuation rate due to adverse events noted with fluvastatin therapy is not significantly distinguishable from the rate associated with placebo. PMID- 8729587 TI - The patient at risk: who should we be treating? AB - Both the European Atherosclerosis Society and the US National Cholesterol Education Program have issued revised guidelines for the prevention of coronary heart disease (CHD), based on a multitude of recent epidemiological and angiographic studies. Both authorities agree that a target plasma low-density lipoprotein cholesterol (LDL-C) level is the single most important parameter, this target level being different for primary and secondary prevention. The introduction of statins for the treatment of hypercholesterolaemia provides an important tool to enable target LDL-C levels to be reached in most cases of primary prevention. For secondary prevention, however, the target LDL-C levels- 2.6 mmol/l (100 mg/dl)--may be achieved in only a fraction of cases. Others may require the concomitant administration of other cholesterol-lowering drugs, such as bile-acid sequestrants (resins) and/or derivatives of fibric acid (fibrates). The use of statin-fibrate combinations has been discouraged since the report by the US Food and Drug Administration of 12 sporadic cases of myositis or rhabdomyolysis. During the past 7 years, however, 21 clinical trials have examined the efficacy and safety of statin-fibrate combinations in a total of 486 patients with a variety of dyslipidaemias. Overall, the combinations were proven to be effective and safe, and the incidence of abnormalities in liver function tests and levels of creatine kinase (CK) was low. A double-blind study has been carried out at the Hadassah University Hospital to examine the efficacy and safety of fluvastatin when combined with either cholestyramine (group 1) or bezafibrate (group 2) for the treatment of 38 patients with heterozygous familial hypercholesterolaemia (FH). Patients in group 2 showed a reduction in plasma LDL C levels of 35% and in LDL-C to high-density lipoprotein cholesterol (HDL-C) ratio of 45% compared with 32% and 38% respectively in group 1. Both cholestyramine and bezafibrate produced an additional benefit of a 13% reduction in LDL-C levels in comparison with fluvastatin as monotherapy. An open-label ongoing study on a larger cohort of FH patients reveals that a decrease in plasma LDL-C levels of up to 38.5% may be achieved with a combination of fluvastatin 80 mg/day and bezafibrate 400 mg/day. In both studies, biochemical safety analyses revealed no notable abnormalities in liver function tests or levels of CK. It was concluded that fluvastatin-bezafibrate is a very effective synergistic therapy for heterozygous FH and is superior to a fluvastatin-cholestyramine combination. PMID- 8729588 TI - Fluvastatin in combination with other lipid-lowering agents. AB - Fluvastatin, a new synthetic inhibitor of HMGCoA (3-hydroxy-3-methylglutaryl coenzyme A) reductase, has been studied in several models to examine its effects when used in combination with other lipid-modifying agents such as derivatives of fibric acid (bezafibrate), resins (cholestyramine), and niacin. The combination of fluvastatin with bezafibrate has been studied in a double-blind trial involving patients with well-documented familial hypercholesterolaemia. Fluvastatin 40 mg/day, combined with either bezafibrate 400 mg/day or cholestyramine 8 g/day, resulted in reductions in levels of low-density lipoprotein cholesterol (LDL-C), these being indistinguishable between the groups; however, significantly greater increases in levels of high-density lipoprotein cholesterol (21.3%) and reductions in levels of triglycerides (25.1%) were seen with the fluvastatin-bezafibrate combination. No notable increases were seen in levels of serum creatine kinase, aspartate aminotransferase, or alanine aminotransferase, and no cases of myopathy were observed. In a study model that examined low-dose combinations of fluvastatin with cholestyramine, reductions in levels of LDL-C of 15.8% and 19.3% were seen with fluvastatin 10 mg and 20 mg, respectively. After an 8-week interval in which a daily dosage of cholestyramine 8 g was added, from baseline, reductions of 26.3% in the 10 mg fluvastatin cholestyramine group and 31.2% in the 20 mg fluvastatin-cholestyramine group were observed, whereas the placebo-cholestyramine group displayed a reduction of 14.9%. Doubling the resin dosage to 16 g/day for the final 8 weeks of the study provided little additional benefit. Myotoxicity has been observed when lovastatin is coadministered with niacin, and so the combination of niacin with fluvastatin has also been studied to examine the possibility of this effect occurring. Patients were randomised to either fluvastatin 20 mg or placebo for 6 weeks, after which time open-label niacin was administered to all patients and titrated to a final dosage of 3 g/day. After 6 weeks, fluvastatin produced a 20.8% reduction in LDL-C levels from baseline. When combined with niacin, a 43.7% reduction was noted at the week 15 endpoint, against the 26.5% reduction seen with niacin monotherapy. The combination was well tolerated, with no reports of myopathy or of significant elevations in creatine kinase or liver transaminase levels. Combinations of fluvastatin with a variety of other agents have been shown to have significant effects on lipid profiles, with no evidence to date of clinically remarkable safety findings. Thus, the use of combination therapies may result in optimal management of patients with moderately severe hypercholesterolaemia and mixed dyslipidaemic profiles. PMID- 8729589 TI - Risk factor management: the cardiologist's perspective. AB - Risk factors are associated with disease and are not necessarily causative; as they are additive they should not be judged in isolation. Thus, 'mild blood pressure' does not imply a 'mild' risk in a cigarette-smoking, hypertensive diabetic. Cardiologists are usually fortunate, in that they see patients when there is already evidence of disease (ie, when secondary prevention is the issue, which is much less contentious than primary prevention). In the 'whizz-bang' specialty of cardiology, a balloon can remove in a matter of seconds what a lipidologist has toiled over for years. Attention to detail may not come easily in a stressful world: it is the detail of prevention that is likely to slow the progression of disease and induce regression rather than angioplasty or bypass surgery. In the presence of coronary artery disease, the essence of management is the team approach, the patient being the most important member of the team. Furthermore, the factors that need the most concentrated effort include cigarette smoking, hypertension, and hyperlipidaemia. As risk factors are additive, treatment of one should not make another one worse. Some drugs for hypertension can exacerbate hyperlipidaemia or impair glucose tolerance (eg thiazides), while ACE inhibitors can beneficially decrease insulin resistance in diabetes. Therapy must be tailored to the individual, but the overall prognostic benefits of drugs that might have slightly adverse metabolic profiles (such as beta-blockers) must not be ignored. Science is often a loser when faced with the power of marketing. When one risk factor (eg hypertension) is present, it is important to be vigorous in treating other factors (eg hyperlipidaemia) with general advice and support, as well as with specific drug therapy if indicated. There are two individuals that tend to lack education in risk factors--the patient and the cardiologist. The patient is a victim of poor investment in prevention, while the cardiologist is put off by a perceived lack of balance from the medical protagonists. PMID- 8729590 TI - Fluvastatin for the prevention of restenosis after coronary balloon angioplasty: angiographic and methodological background of the fluvastatin angioplasty restenosis trial. AB - Luminal renarrowing (restenosis) is the major limitation of percutaneous transluminal coronary angioplasty (PTCA), and the search for a 'magic bullet' to prevent this apparent biological healing response to vessel injury has thus far been unsuccessful. Large clinical trials using serial quantitative coronary angiography have, however, provided some valuable insight into this area. In particular, the restenosis process may be measured as the loss in minimal luminal diameter from post-PTCA to follow-up angiography, and is essentially ubiquitous and normally distributed. The angiographic outcome of clinical trials can thus be appropriately evaluated using a continuous rather than a categorical approach, which also considerably reduces the number of patients required. Fluvastatin, a synthetic 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor, has been shown experimentally to reduce the neointimal proliferative response after PTCA, independent of its lipid-lowering action. The FLuvastatin Angioplasty REstenosis trial was designed to evaluate whether fluvastatin 40 mg twice daily, commencing at least 2 weeks before planned PTCA, can reduce luminal loss by 30% from successful PTCA to follow-up angiography at 26 +/- 2 weeks in 730 evaluable patients. PMID- 8729592 TI - Medicine and the media--education, not exploitation. PMID- 8729591 TI - Lipoprotein(a) as a marker for coronary heart disease. AB - Lipoprotein(a) (Lp(a)) resembles a low-density lipoprotein (LDL) particle, but with the addition of apolipoprotein(a). Apolipoprotein(a) (apo(a)) is homologous to plasminogen, and its similarity to plasminogen indicates a prothrombogenic role for Lp(a), whereas the similarity of Lp(a) to LDL suggests a proatherogenic role. Thus, Lp(a) is a link between atherosclerosis and thrombosis. The apo(a) gene is highly polymorphic, and a relationship exists between the different apo(a) alleles and plasma Lp(a) levels. There is also evidence to suggest an association between serum Lp(a) concentration and the frequency of cardiovascular atherosclerosis, particularly myocardial infarction in men, although some of the recently published epidemiological studies have been controversial. The heightened awareness of increased Lp(a) levels being a risk factor for coronary artery disease has led to the evaluation of the effects of lipid-lowering agents on Lp(a) levels. The data that have emerged, however, are conflicting, and are derived mostly from short-term studies involving the use of statins. Our group has studied the short- and long-term effect of fluvastatin on Lp(a) in hypercholesterolaemic patients; fluvastatin was found to have no effect in such patients as observed in the short-term studies, but significant reductions in Lp(a) levels were noted during long-term treatment. Although our findings are in agreement with those of a previously published report, there is still a great need to confirm the data that have been reported in separate studies. PMID- 8729593 TI - HRT use: where do we stand today? PMID- 8729594 TI - What factors influence general practitioners' commencement of hormone replacement in perimenopausal women? AB - The purpose of this study was to determine factors influencing general practitioners' (GPs) commencement of hormone replacement (HRT) in perimenopausal women referred for bone mineral density analysis. GPs who referred 80 untreated healthy perimenopausal women to a Western Sydney teaching hospital for osteoporotic fracture risk assessment were surveyed. All patients underwent dual energy X-ray absorptiometry of the lumbar spine, femoral neck and distal radius. Subjects were reported as 'at risk of fracture' if their bone mineral density was more than two standard deviations below the mean of young adults. Patients had normal (n = 55) or reduced (n = 4) bone mineral density at all sites; 21 subjects (26%) had regional osteoporosis. HRT was commenced in 35 women. A multivariate logistic model showed that menopausal symptoms (OR 173.9, p < 0.001) and reduced bone mineral density at any site (OR 28.3, p < 0.001) were highly significant independent factors predicting HRT (R2 0.48). Other clinical features such as low weight, family history and smoking status did not contribute further to GP decision making. PMID- 8729595 TI - Nocturnal 8 mg dexamethasone suppression test: a practical and accurate test for identification of the cause of endogenous Cushing's syndrome. AB - The efficacy of a nocturnal 8 mg dexamethasone suppression test (nocturnal DST) was compared with that of the standard high-dose dexamethasone suppression test (standard DST) in identifying the cause of endogenous Cushing's syndrome in 10 proven cases with Cushing's disease, 20 with adrenal tumours, and one with ectopic ACTH syndrome. The nocturnal test compared serum cortisol concentration at 8 am before and after administration of a single dose of 8 mg dexamethasone at 11 pm. Suppression of serum cortisol level to < 50% of the baseline value indicated a diagnosis of Cushing's disease, while a lack of suppression below that limit indicated one of the other two causes of Cushing's syndrome: glucocorticoid-secreting adrenal tumour or ectopic ACTH syndrome. The nocturnal DST had a sensitivity of 90%, a specificity of 100%, an accuracy of 96.8%, a positive predictive value of 100%, and a negative predictive value of 95.5%. These values are comparable to the efficacy of the standard DST in distinguishing Cushing's disease from glucocorticoid-secreting adrenocortical tumour or ectopic ACTH syndrome. Furthermore, this rapid test does not require hospitalisation or urine collection like the standard DST. The nocturnal 8 mg dexamethasone suppression test is practical, fairly reliable, and an effective alternative with which to identify the cause of endogenous Cushing's syndrome. PMID- 8729596 TI - Patient perception of the diskus inhaler: a comparison with the turbuhaler inhaler. AB - Asthmatic patients who were regular and experienced users of the metered-dose inhaler (MDI) but with no history of powder inhaler use were interviewed by questionnaire. Most (71%) of the 159 adults recruited had suffered from asthma for more than five years; 89% had used an MDI for more than one year. In all, 153 (97%) found their current MDI easy or very easy to handle. Effectiveness in delivering the drug and ease of operation were the two features of the device they were most satisfied with. The two features 'of an ideal inhaler' rated most highly were the presence of a dose counter and ease of use during an attack (58% and 72% of respondents, respectively). Other important features included hygiene, sensation that a dose had been taken, and small size. When attitudes towards the Diskus inhaler and the Turbuhaler inhaler were compared, 108 (68%) would have been quite or very happy to have a Diskus inhaler prescribed for them, and 79 (50%) a Turbuhaler inhaler. The Diskus inhaler scored significantly better than the Turbuhaler inhaler for knowing how many doses were left, the presence of an attached cover, attractiveness (p < 0.001), hygiene, shape, weight (p = 0.001), and perceived ease of use (p = 0.002). The Turbuhaler inhaler scored significantly better in terms of size (p < 0.001). Overall, the Diskus inhaler was preferred by 104 of 159 patients (p < 0.001), with perceived ease of use, the dose counter, and shape cited as the main reasons for preference. PMID- 8729597 TI - A survey of peripheral oedema in elderly patients admitted to a geriatric ward. AB - Peripheral oedema is common in elderly people. This two-month survey of an acute geriatric ward found that 48% of the 63 admissions over that period had oedema either on, or during the course of their admission. In many cases this was mild and asymptomatic, but complications of oedema were the cause of admission for four patients. The oedema was often multifactorial, but gravitational forces in dependent limbs seemed to be an important factor for many cases. Few patients were receiving any specific treatment for their oedema. It is concluded that oedema may be underrecognised, and that if greater attention is paid to this problem at an earlier stage, some of the associated morbidity could be avoided. PMID- 8729598 TI - Critical drug appraisal: etidronate intermittent cyclic therapy for postmenopausal osteoporosis. AB - An overview of intermittent cyclical therapy with oral etidronate disodium in the treatment of postmenopausal osteoporosis is given. Etidronate has been used successfully in the treatment of Paget's disease and for the prevention and treatment of heterotopic ossification after hip replacement and spinal cord injury. It is approved for the treatment of postmenopausal osteoporosis in many European countries, Canada and Australia. Studies of such therapy indicate that etidronate significantly increases bone mass, reduces the risk of future vertebral fracture, and is safe and generally well tolerated. PMID- 8729599 TI - The epidemiology of osteoporosis. AB - Osteoporosis is a skeletal disorder characterised by low bone mass and an increased susceptibility to fracture. The last two decades have seen an increase in our understanding of the epidemiology of fractures at the three most frequent sites (the hip, wrist and vertebral body). These new insights have led to the delineation of preventive strategies against these fractures for both the general population, and those individuals at highest risk. This review covers these descrptive epidemiological characteristics, as well as approaches to prevention. PMID- 8729600 TI - HRT, a general practice approach: how to reach the most vulnerable. AB - Hormone replacement therapy has been shown to be effective in the preservation of bone density and its use should lead to a substantial reduction in the fracture rate of older women. Observational studies have also suggested that it may lead to a reduction in the incidence of coronary heart disease. Both effects are relevant to preventive care in general practice and have been applied within the framework of health promotion clinics. However, the tendencies for patients who are already healthy to select themselves for attendance at these clinics, and for long-term hormone therapy, mean that efforts to improve the health of the population do not reach those most in need. Groups most at risk should be defined by individual practices and identification of high-risk patients be followed by appropriate investigation and treatment. It is possible to identify the following high-risk groups: long-term users of corticosteroid therapy or thyroxine; women who have had an early hysterectomy or oophorectomy; women with known coronary artery disease, diabetes or familial hyperlipidaemia. Methods of ascertainment and investigation are described, as are the various modes of therapy and lifestyle modifications considered appropriate. Health promotion clinics are not currently reaching those most in need, and preventive therapy should be extended to target the most vulnerable. PMID- 8729601 TI - Prevention of both osteoporosis and coronary heart disease in primary care. AB - This review describes the importance of both osteoporosis and CHD as causes of morbidity and mortality, and emphasises the commonality of risk factors for the development of both conditions. On the basis of this information, an approach to preventing both diseases is outlined--population-based and personal, as delivered by primary care teams. HRT is seen to have a key role, but the importance of tackling lifestyle issues is also stressed. PMID- 8729602 TI - Comment: preventive strategies: is current clinical practice effective for bones? PMID- 8729603 TI - Tuberculous mastitis: a rare disease. AB - Two cases of tuberculous mastitis are described. This disease remains rare. While routine laboratory investigations are not helpful in its diagnosis, fine needle aspiration to allow histologic examination of tissue and culture is essential- both to differentiate tuberculous mastitis from pyogenic breast abscess and breast carcinoma, and to guide antimicrobial therapy. Antituberculous drugs in combination with aspiration or surgical drainage are usually associated with an excellent outcome. PMID- 8729604 TI - Changes in the use of EEG following clinical audit. AB - An audit of the clinical use of EEG in mentally handicapped patients was performed over a three-year period. EEG requests seemed inappropriate in more than one-quarter of cases. Changes in clinical practice during the 10 months following presentation of these audit results were examined. There was a marked reduction in EEG requests, which were subsequently used in a more cost-effective and clinically appropriate way. There was also a reduction in the time taken to process requests. PMID- 8729605 TI - Diaphragmatic herniation of the large bowel associated with unusual clinical and electrocardiographic presentation. AB - This is the first case reported of combined hiatus hernia of the stomach and herniation of the splenic flexure of the colon through the oesophageal hiatus. It was associated with unusual clinical and electrocardiographic findings. We postulate that these were the result of an increase in intrathoracic pressure induced by the herniating structures. PMID- 8729606 TI - Musical hallucinations associated with recurrence of a right occipital meningioma. AB - The case of an 84-year-old woman who experienced musical and visual hallucinations after recurrence of a right occipital meningioma is reported. PMID- 8729607 TI - Dyspareunia in undiagnosed Sjogren's syndrome. AB - A case of superficial dyspareunia in a 46-year-old women is described. Clinical suspicion, examination and investigations supported a diagnosis of Sjogren's syndrome. PMID- 8729608 TI - Bouveret's syndrome (gallstone ileus)--a minefield. AB - Gallstone ileus is an uncommon cause of intestinal obstruction. Diagnosis may be delayed or even missed. This case history illustrates some characteristics of gallstone ileus that can complicate its subsequent management. PMID- 8729609 TI - Progressive supranuclear palsy presenting with psychiatric features. AB - A patient with progressive supranuclear palsy who presented with psychiatric features is reported. His case illustrates the difficulty of early diagnosis of this condition. Associated psychiatric symptoms are common and may precede the occurrence of gaze palsy. Our patient's behavioural problems responded to fluoxetine. PMID- 8729610 TI - Tuberculous parotitis: two cases in Libyan patients. AB - Two patients with tuberculous parotitis are described. This condition is rare, and has not previously been recorded in Libya. Unilateral tuberculous parotitis is clinically indistinguishable from a parotid tumour, and imaging techniques may also fail to reveal the true diagnosis. If other features point to tuberculous parotitis, fine needle aspiration allowing bacteriological confirmation is the most useful investigation. PMID- 8729611 TI - Multiple tumours of the salivary glands--terminology and nomenclature. AB - Multiple tumours of the salivary glands are very rare and their combinations according to histological classification of the tumours, localisation and origin (origin in independent topographical areas or in the same tissue) are diverse. The following two categories can be distinguished: common occurrence of multiple salivary gland tumours with identical histology, or with different histology. In either group the tumours can be unilateral or bilateral, synchronous or metachronous. The most common multiple tumours with an identical histology are Warthin tumours and pleomorphic adenomas. Bilateral occurrence has been observed especially in oncocytomas, acinic cell carcinomas and basal cell adenomas. In the group of multiple tumours with differing histology, Warthin tumours and pleomorphic adenomas show a number of combinations with other adenomas or carcinomas of the salivary glands. Notable also is the simultaneous occurrence of salivary gland tumours with other oral tumours or extraglandular tumours, especially thyroid carcinomas and breast carcinomas. Multiple salivary gland tumours must be distinguished by nomenclature from tumours with biphasic differentiation and hybrid tumours. Tumours with biphasic differentiation are defined as regular, recurring mixtures of two cellular components in the same tumour and have a corresponding term in the tumour classification. Hybrid tumours are very rare and are composed of two different tumour entities within the same topographical area. Each of the tumour entities conforms with an exactly defined tumour category. PMID- 8729612 TI - Identification of basal lamina acidic glycoconjugates, particularly heparan sulphate proteoglycans, using a poly-L-lysine-gold probe in induced oral carcinomas. AB - Acidic glycoconjugates represent the major non-fibrous macromolecular components that form the extracellular and cell-associated matrices of all animal tissues. The constituent molecules are principally structural glycoproteins and proteoglycans. While their protein component is determined by gene pools, it is the polyanionic (acidic) nature of the polysaccharides, determined by their degrees of carboxylation and sulphation, which confers both functional and diagnostic status on these molecules. Sulphated glycoconjugates in the basal laminae have been reported to play a role in tumour invasion and metastasis. In this study, we used cationic colloidal gold together with transmission electron microscopic methods to compare the expression of acidic glyconconjugates in the basal lamina of both normal rat tongue mucosa and experimentally induced oral carcinomas. Results indicated that heparan sulphate rich glycoconjugates were predominant and were mostly confined to the lamina lucida of the basal lamina in normal oral mucosa. Conversely, observation of basal laminae associated with induced carcinomas showed less intense and more widely dispersed gold labelling for heparan sulphate. The observed differences in gold labelling may reflect modified metabolism of sulphated glycoconjugates or result from the action of degradative enzymes in the induced tumours. PMID- 8729613 TI - Expression of different keratins in salivary gland tumours. AB - Twenty-four salivary gland tumours (six pleomorphic adenomas, two myoepitheliomas, five basal cell adenomas, six adenoid cystic carcinomas and five polymorphous low grade adenocarcinomas) were investigated by an immunocytochemical technique using monoclonal antibodies against cytokeratins (CKs) 7, 8, 10, 13, 14, 18 and 19. The luminal cells of ductal structures of the tumours reacted with all the CKs studied except for CK 13 and CK 10 and sometimes CK 14, showing an immunoprofile comparable to that of the intercalated segment of a normal salivary gland. The outer cells of the ducts rarely stained with CK 14, confirming that full differentiation of the myoepithelial cells is seldom achieved in tumours. Considerations were made regarding the intriguing expression of CK 14, the heterogeneous expression of CKs in the modified myoepithelial cells and the immunoprofile of the polymorphous low-grade adenocarcinoma. PMID- 8729614 TI - Treatment of non-Hodgkin's lymphoma of Waldeyer's ring: radiotherapy versus chemotherapy versus combined therapy. AB - Treatment of stage IA non-Hodgkin's lymphoma (NHL) of Waldeyer's ring remains controversial, probably because of the small number of patients and the scarcity of controlled studies. Between 1981 and 1991, 316 patients with stage I NHL of Waldeyer's ring were randomised for treatment with radiotherapy alone (extended fields), 101 patients; combined chemotherapy with a regimen of CHOP (cyclophosphamide, vincristine, doxorubicin, and prednisone) or CHOP-like (epirubicin instead of doxorubicin), 106 patients; and combined therapy (radiotherapy followed by the same combination chemotherapy), 109 patients. Median follow-up was 6.8 years. Complete response was achieved in 93, 87 and 97%, respectively. Relapses were least frequent in patients treated with combination therapy. The 5-year rate for failure-free survival was 48% for radiation therapy, 45% for the patients who were treated with chemotherapy, which was statistically significantly less than the 83% for patients treated with combined therapy (P < 0.001). Overall survival was also better in the combined therapy arm: 90%, statistically different to 58% for the patients treated with chemotherapy alone and 56% for patients treated with radiation therapy (P < 0.001). Toxicity was mild and late side-effects were not observed in any patients. From these results combined therapy should be considered as the best therapeutic approach in patients with localised NHL of Waldeyer's ring. PMID- 8729615 TI - The relationship between vascularity and cell proliferation in human normal and pathological lesions of the oral cheek epithelium. AB - The present study investigates relationships between neovascularisation and PCNA cell proliferation markers in different pathological lesions of the oral cheek mucosa. All specimens were fixed in 10% formalin and routinely processed for histology. Six normal (N) samples were taken from resection margins of benign lesions. The pathological lesions consisted of chronic inflammation (n = 10), lichen planus (n = 7), fibrous hyperplasia (n = 11), dysplasia (n = 5), squamous cell carcinoma (n = 22) and epithelium adjacent to carcinoma (n = 6). Adjacent 5 microns sections were stained with monoclonal antibodies against vimentin (clone no. V9) for identification of stromal blood vessels and against proliferating nuclear antigen (PCNA/PC10) using ABC immunoperoxidase techniques. Point counting was used to obtain the primary morphometric data using a Zeiss VIDAS image analyser. No attempt was made to classify the different types of blood vessels. The morphometric blood vessel parameters estimated were volume density, number per unit area, length per unit volume and mean transverse sectional area. PCNA indices were determined by estimating the percentage frequency of PCNA positive nuclei in basal and spinous strata. Generally, there were significant increases in all PCNA indices and blood vessel parameters between the N group and the different pathological lesions. A highly positive correlation was detected between all PCNA indices and blood vessel parameters. These data suggest that increased vascularity and angiogenesis occur in support of actively proliferating and transforming oral epithelial cells in order to permit growth. PCNA indices and blood vessel parameters may have a potential application as diagnostic and prognostic indicators. PMID- 8729616 TI - Lectin histochemistry of oral premalignant and malignant lesions: correlation of JFL and PNA binding pattern with tumour progression. AB - The expression of glycoconjugates specific to Jack fruit lectin (JFL) and peanut agglutinin (PNA) in various clinicopathological stages of tumour progression in the oral mucosa were studied. These included various clinical forms of dysplastic and non-dysplastic oral leucoplakias, carcinomas, normal keratinising (gingiva) and non-keratinising (buccal mucosa) epithelia. It was seen that the binding patterns of PNA and JFL in the epithelial cells of various types of oral lesions were more or less similar. Normal non-keratinising epithelium showed mild membrane staining only in the spinal layers, while normal keratinising epithelium showed a moderate membrane staining and mild cytoplasmic staining in all layers. Moderate membrane and mild cytoplasmic staining were observed in leucoplakias, irrespective of various clinical or histological types. In carcinomas, the intensity of lectin binding was high, particularly in the membrane of differentiated cells. Correlation analysis of the binding pattern of PNA and JFL showed significant correlation in the membrane and cytoplasm of all layers with histological stages of tumour progression. The present study thus showed that PNA and JFL may be used as cytochemical probes in differentiating malignancy from benign lesions of the oral mucosa. PMID- 8729617 TI - B-cell monoclonality in salivary lymphoepithelial lesions. AB - It is well recognised that lymphoma may arise in a lymphoepithelial lesion of the salivary glands. Although the histological features of this lesion are well described, it is not clear what proportion contain monoclonal populations of lymphocytes at outset. In this study, 22 routinely processed lymphoepithelial lesions in parotid glands were examined for B-cell monoclonality using the polymerase chain reaction (PCR) to amplify the immunoglobulin heavy chain gene and using in situ hybridisation or immunohistochemistry to detect kappa or lambda light chain restriction. B-cell monoclonality was identified in 17/22 (77.3%) cases using a combination of the three methods. The detection rate for B-cell monoclonality was highest using PCR with 15/22 (68%) cases containing monoclonal immunoglobulin heavy chain gene rearrangements. In a proportion of cases the results of in situ hybridisation and immunohistochemistry were judged to be inadequate and this was probably a reflection of variations in fixation. In 7 patients, sequential biopsies were available from other sites and 6 of these also showed B-cell monoclonality. The results confirm the high prevalence of B-cell monoclonality in lymphoepithelial lesions of the major salivary glands. Furthermore, these results would suggest that PCR is a more reliable technique to identify B-cell monoclonality in routinely processed lymphoepithelial lesions compared to in situ hybridisation and immunohistochemistry. PMID- 8729618 TI - p53-HSP70 complexes in oral dysplasia and cancer: potential prognostic implications. AB - We have previously shown overexpression of p53 and 70 kDa heat shock protein (HSP70) in potentially malignant, as well as malignant, oral lesions in an Indian population, suggesting that alterations of p53 and HSP70 expression may occur in the early stages of oral tumorigenesis. Herein we report immunological evidence for the specific association between p53 and HSP70 in potentially malignant and malignant oral lesions. This association was indicated by coimmunoprecipitation of p53 and HSP72/73 proteins observed with either an anti-p53 monoclonal antibody or an anti-HSP72/73 antibody. Furthermore, reciprocal blotting analysis showed that HSP72/73 proteins did not share an epitope with p53, confirming that the coimmunoprecipitation of p53 and HSP72/73 is a physical association of the proteins in potentially malignant lesions (dysplasia) and oral squamous cell carcinomas (SCCs). p53-HSP70 complex formation was observed in 19/52 cases of oral SCCs and 10/53 cases of potentially malignant lesions (leucoplakia). Normal oral mucosa did not show the presence of p53-HSP70 complexes (0/20 cases). p53 HSP70 complex formation may be one of the mechanisms of stabilisation of p53 protein resulting in its increased levels in potentially malignant and malignant oral lesions and may be implicated in oral carcinogenesis. PMID- 8729619 TI - In vitro uptake of bromodeoxyuridine by human nasopharyngeal carcinoma (NPC) and its relation to clinical findings. AB - A cell kinetic study of 27 newly diagnosed patients with nasopharyngeal carcinoma (NPC) using the in vitro bromodeoxyuridine (BrdU) technique was performed. The results were reproducible as demonstrated by three independent sections performed on each patient. No correlation between BrdU labelling index (LI) and Ho's clinical staging was found. A higher LI was associated with the development of distant metastases (P = 0.057). Statistically significant correlation was found between low LI and longer duration required to achieve complete remission in the primary site of disease (P = 0.026). This study suggests a potential role for in vitro BrdU labelling index as a prognosticator for NPC prior to treatment. PMID- 8729620 TI - Age-dependence of human papillomavirus DNA presence in oral squamous cell carcinomas. AB - The aetiology of oral cancer is thought to be multifactorial. Apart from the two known major risk factors (tobacco and alcohol), a viral aetiology has been proposed, with special reference to human papillomavirus (HPV). 35 cases of oral squamous cell carcinoma (OSCC), seen at the Departments of Oral & Maxillofacial Surgery and Oral Pathology and Otolaryngology of the Free University of Amsterdam, were analysed as well as 12 biopsies of clinically and histologically normal gingival mucosa collected from healthy individuals after tooth extractions, using the polymerase chain reaction (PCR) and two different sets of primers that are able to detect a broad spectrum of HPV types. An overall HPV positivity of 54.3% in OSCC was found, the majority of positive cases (78.9%) harbouring HPV type 16. In contrast, no positivity for HPV was detected in the clinically normal oral mucosal samples analysed. Furthermore, a significant association between HPV presence and age was found: patients older than 60 years showed a lower prevalence of the virus (29.4%) compared with patients below this age (77.8%) (P < 0.05). The results from the present study suggest an association between HPV and OSCC, particularly in patients under the seventh decade. PMID- 8729621 TI - Is oral cancer susceptibility inherited? Report of five oral cancer families. AB - All the oral cancer patients registered at the Regional Cancer Centre, Trivandrum, during January to July 1995 were subjected to detailed pedigree analysis. This revealed that oral cancer tends to aggregate in families. Like other familial cancers, a family history of oral cancer was associated mostly with an early age of onset of the disease. Family members without habits such as tobacco chewing, smoking or alcohol consumption were also affected. These observations prompt us to suggest the probable inheritance of an oral cancer susceptibility gene in these families. The familial aggregation, mostly site specific, with an autosomal dominant mode of inheritance, was observed in 0.94% of the total oral cancers. This necessitates the need to undertake studies to elucidate the molecular lesions responsible for oral cancer susceptibility in families. PMID- 8729622 TI - Kaposi's sarcoma herpesvirus in oral Kaposi's sarcoma. PMID- 8729623 TI - Remotely sensed surrogates of meteorological data for the study of the distribution and abundance of arthropod vectors of disease. AB - This paper gives an overview of how certain meteorological data used in studies of the population dynamics of arthropod vectors of disease may be predicted using remotely sensed, satellite data. Details are given of the stages of processing necessary to convert digital data arising from satellite sensors into ecologically meaningful information. Potential sources of error in these processing steps are also highlighted. Relationships between ground-measured meteorological variables (saturation deficit, ground temperature and rainfall) and data from both the National Oceanic and Atmospheric Administration's, polar orbiting, meteorological satellites and the geostationary, Meteosat satellite are defined and examples detailed for Africa. Finally, the current status of existing satellite platforms and future satellite missions are reviewed and potential data availability discussed. How such satellite-based predictions have proved valuable in understanding the distribution of tsetse fly species in Cote d'Ivoire and Burkina Faso will be the subject of a future review. PMID- 8729624 TI - Treatment of acute, uncomplicated, falciparum malaria with oral dihydroartemisinin. AB - A clinical trial of oral dihydroartemisinin for the treatment of acute, uncomplicated, falciparum malaria involved 53 adult patients in Thailand. Each received a total dose of 480 mg over 7 days (120 mg given immediately, followed by 60 mg/day) after being admitted to the Hospital for Tropical Diseases in Bangkok for 28 days. Most (92%) completed the 28-day follow-up but four patients left the hospital early, for reasons unrelated to their treatment. Most patients showed clinical improvement 1-3 days after starting treatment and none suffered from serious adverse reactions. The cure rate was 90% (44/49). The mean (S.D.) parasite-clearance time was 40.4 (14.1) h and the mean fever-clearance time was 37.0 (30.2) h. Seven patients had a brief increase in parasitaemia after initiation of treatment but subsequent counts declined dramatically. Five patients who failed treatment (RI response) were successfully treated with quinine plus tetracycline for 7 days. No RII or RIII responses were observed. These findings indicate that treatment with oral dihydroartemisinin is effective and well tolerated, and that dihydroartemisinin may be suitable as an alternative treatment for acute, uncomplicated, falciparum malaria. Comparisons with other conventional antimalarial drugs in a large, double-blind, randomized trial and studies of dihydroartemisinin in combination with other, long-acting antimalarials are needed. PMID- 8729625 TI - Malaria transmission by Anopheles punctulatus in the highlands of Irian Jaya, Indonesia. AB - A 21-month (1991-1992) assessment of malaria and the relative contribution of Anopheles punctulatus and An. koliensis to its transmission was conducted in Oksibil, a highland community in Irian Jaya, Indonesia. Nearly all (98%) of the 2577 Anopheles from indoor or outdoor human-landing collections were An. punctulatus. Nineteen (0.75%) of the 2518 An. punctulatus tested were positive when tested for malaria circumsporozoite protein (CSP) by ELISA: nine (45% of positives) for Plasmodium vivax-210, six (30%) for Plasmodium falciparum, three (15%) for P. vivax-247 and two (10%) for P. malariae. One mosquito was positive for both P. vivax 210 and P. falciparum. All 59 An. koliensis tested were negative. All mosquitoes positive for CSP were from indoor collections, 89% from the main houses and the remaining 11% from the traditional dwellings (bokams). These findings are discussed with respect to highland malaria epidemiology and transmission in the Oksibil Valley. PMID- 8729626 TI - The economic burden of lymphatic filariasis in northern Ghana. AB - The socio-economic impact of lymphatic filariasis was investigated in a rural community in northern Ghana, as part of a multi-country study. The entire community was followed up for a period of one full calendar year, to document episodes of acute adenolymphangitis (ADL), treatment seeking and the cost of treatment. Cases of chronic filarial disease identified at the start of the study were also visited every 3 months to monitor treatment seeking and the costs of treatment. The subjects, who were mainly subsistence farmers, were found to be materially very poor. The incidence of ADL, which lasted an average of 5 days, was found to be very high. Most of those affected were unable to perform their normal activities for much of the ADL episode. In general, most subjects with long-standing chronic disease, such as elephantiasis of the leg and hydroceles, tended not to seek treatment except when there was superimposed ADL. Some aspects of treatment seeking, the cost of treatment, the burden of the disease on the community, and the public-health implications are discussed. PMID- 8729627 TI - Intestinal parasites of humans in a highland community of Irian Jaya, Indonesia. AB - An indigenous, highland community in the Oksibil Valley of Irian Jaya, Indonesia, was examined on two occasions to determine the prevalence of intestinal parasites. Overall, 478 subjects from three villages in the study area, about 10% of the available population, were sampled. Using a standard, wet-mount technique, 15 different species of parasites were found. Hookworm was predominant in all age groups, with prevalences of 51%-86%, followed by Ascaris lumbricoides (46%-57%) and Trichuris trichiura (15%-25%). Strongyloides stercoralis was only seen infrequently (< 1%). In general, intestinal protozoa were uncommon. Entamoeba histolytica (14%-20%) and Entamoeba coli (12%-17%) were the most frequent. Sex- and age-specific differences were not dramatic but prevalences and species diversity both increased with age up to 6-15 years, then decreased slightly into adulthood. Rarely were faecal samples free of detectable parasites and some adult subjects had as many as six species. The results are discussed with respect to previous studies in the highlands of New Guinea and the local transmission of soil- and water-borne parasites. PMID- 8729628 TI - Trichuris trichiura infection and cognitive function in Jamaican school children. AB - Trichuris trichiura is extremely prevalent worldwide and there is concern that this geohelminth may affect the cognitive function of children in developing countries. A random-controlled, double-blind, treatment trial was conducted in Jamaican children with light to moderate infections. This was part of a research programme involving several studies in Jamaica. Ninety-seven subjects, each with a minimum of 1200 T. trichiura eggs/g faeces, were randomly assigned to placebo (N = 48) or treatment (N = 49) groups. Each pair of infected children was matched with an uninfected classmate (N = 48). All children were given seven cognitive function tests: French-learning; digit spans (forwards and backwards), Corsi block span; fluency; picture search; and silly sentences. Albendazole was given to the treatment group and the other groups received a placebo. Three months later, these treatments were repeated, and the cognitive function battery was given again. On pre-test, the infected groups performed significantly poorer only in the silly-sentence test (analysis of variance F-value = 8.17; two degrees of freedom; P < 0.001). There was no significant improvement with treatment in any of the tests. Taking into account these findings and those of the other Jamaican studies, it is therefore probable that light to moderate Trichuris infections have little effect on cognitive functioning in school children who have adequate nutritional status. PMID- 8729630 TI - Ovulation and post-ovulation changes in the terminal region of the ovarioles of the mosquito Aedes aegypti (L.) (Diptera: Culicidae). AB - Detachment of the follicular epithelium from the chorion occurs prior to the passage of the mature egg into the calyx lumen, resulting in a mature oocyte lying free in an epithelial sac. The mature oocytes rupture the epithelial sac, including the tunica propria at the posterior pole, then break through the basal (or granular basal) bodies to enter the calyx lumen. Just after ovulation, the formerly terminal parts of the ovariole remain attached to the posterior end of the egg sac near its opening. The terminal parts then undergo changes in conjunction with complete degeneration of the egg sac and development of the basal body. They cannot be used for age determination of females. PMID- 8729629 TI - Prevalence of antibodies against simian immunodeficiency virus (SIV) and simian T lymphotropic virus (STLV) in a colony of non-human primates in Kenya, East Africa. AB - Sera (165 samples in 1988 and 66, follow-up samples in 1989) were collected from olive baboons, African green monkeys, Syke's monkeys and grey mangabeys kept in a semi-free, breeding colony at the Institute of Primate Research (IPR) in Nairobi, Kenya. The levels of antibodies to simian T-lymphotropic virus (STLV) or simian immunodeficiency virus (SIV), and the reactivity patterns of positive sera to various lentivirus subgroup antigens, were then determined. The results of tests using enzyme-immunoassay kits were confirmed by western blots. The prevalence of antibodies which reacted with the Kenyan SIVagm(KEN) isolate was 28% in the African green monkeys tested and 34% in the Syke's monkeys. STLV seroprevalence was 25% in the African greens and 20% in the Syke's. No antibodies to either SIV or STLV were detected in the olive baboons or grey mangabeys. More SIV-positive samples were detected in western blots when SIVagm(KEN) was used as antigen than when SIVagm(CAR014), a geographically distinct isolate from the Central African Republic, was used. However, SIVagm(KEN)-positive sera were more reactive against SIVagm(CAR014) than SIVsmm and SIVmac subgroup antigens, indicating that the two isolates from the African green monkey, CAR014 and KEN, remain antigenetically close even though they were recovered in two geographically distinct regions. To date, no clinical disease has been linked with SIV and STLV infection in the African green or Syke's monkeys in the colony. However, the relatively high prevalence of anti-SIV and anti-STLV antibodies in these monkeys offers an opportunity for prospective studies on the transmission and natural history of both viruses in a single colony. PMID- 8729631 TI - Patterns of health-care utilization and morbidity in a rural community near Lahore, Pakistan. AB - Patterns of health-care utilization and of morbidity were investigated in a demographically defined community: the 1400 inhabitants of a rural village near Lahore, Pakistan. The data collected, by semi-monthly clinic surveys from February 1982 to January 1983, showed that 42% of the study population sought health care during the year-long study period, between one and 10 (mean 2.1) times each. Females attended the clinics twice as often as males. The 1193 person visits that were recorded at the field clinics yielded a total of 1354 cases of various diseases. The cumulative number of clinic-visits per person fitted a negative binomial distribution, indicating that health problems were concentrated in a small portion of the population. When the patients were classified according to the International Classification of Diseases, the most common disease category was that of the respiratory system (27.6%), followed by infectious and parasitic diseases (18.7%), diseases of the skin (13.7%) and those of the nervous system and sense organs (10.9%). Classification of the cases by single disease condition indicated that diarrhoea, helminthiasis, malaria, anaemia, conjunctivitis, bronchitis, coryza, pharyngitis, tonsillitis, dyspepsia and pyoderma were the most common diseases. In view of the fact that infectious diseases continue to be a major public health problem in rural Pakistan, the need for a sound health policy that is primarily focused on preventive medicine, especially health education, is apparent. PMID- 8729632 TI - Recovery of Schistosoma haematobium eggs in venous blood collected for diagnosis of Wuchereria bancrofti infection. PMID- 8729633 TI - Nairobi sheep disease virus isolated from Haemaphysalis intermedia ticks collected in Sri Lanka. PMID- 8729634 TI - Quiescent ovarioles in the mosquito Aedes aegypti (L.) (Diptera: Culicidae). PMID- 8729635 TI - A strategy for validation of bioanalytical methods. PMID- 8729636 TI - Simultaneous quantitative determination of metronidazole and nalidixic acid in tablets by difference spectroscopy. AB - A difference spectrophotometric procedure has been developed for simultaneous determination of metronidazole (MDZ) and nalidixic acid (NA) in tablets. The method comprised the measurement of the absorbance of a solution of the tablet extract in 0.1 M NaOH relative to that of an equimolar solution in 0.1 M HCl at 292 nm for NA and 325 nm for MDZ. The presence of identical isosbestic points for pure drug solutions and tablet extracts indicated the non-interference of excipients in the absorption at these wavelengths. Compliance with Beer's law was observed in the concentration ranges 5-25 micrograms ml(-1) for MDZ and 15-35 micrograms ml(-1) for NA these wavelengths. PMID- 8729637 TI - Long-term stability of 5-fluorouracil stored in PVC bags and in ambulatory pump reservoirs. AB - Prolonged infusions of 5-fluorouracil (5FU) have been used since the early 1960s, but recently there has been a major resurgence of interest, partly because of the advent of electronically controlled portable infusion pumps. Admixtures of new formulation 5FU were subjected to stability studies to establish the feasability of continuous infusions. In the first study, the stability of 5FU, 1 or 10 mg ml( 1), was determined in poly(vinyl chloride) (PVC) bags (0.9% sodium chloride injection or 5% dextrose injection) at 4 and 21 degrees C after storage for 0, 1, 2, 3, 4, 7 and 14 days. In the second study, the stability of undiluted 5FU was tested at different temperatures (4 or 33 degrees C) in ethylene-vinyl acetate (EVA) or PVC ambulatory pump reservoirs after storage for 0, 3, 5, 7 and 14 days. For each condition, samples from each admixture were tested for drug concentration by stability-indicating high-performance liquid chromatography. The admixtures were also monitored for precipitation, colour change and pH. Evaporative water loss from the containers was measured. The stability of 5FU in PVC bags was unaffected by 14 days of storage at either 4 or 21 degrees C. When stored in EVA reservoirs, 5FU was stable for at least 2 weeks at 33 degrees C and for 3 days at 4 degrees C (a precipitate was observed after 3 days). In PVC reservoirs, 5FU was stable for over 14 days at 33 degrees C, but at 4 degrees C a precipitate appeared after 5 days. Loss of water through the reservoirs was substantial only at 33 degrees C for 14 days, and gave falsely high readings. PMID- 8729638 TI - Determination of analytical error function for beta-blockers as a possible weighting method for the estimation of the regression parameters. AB - Three analytical methods have been developed and validated for the quantification of beta-blockers (celiprolol, bisoprolol and oxprenolol) using high performance liquid chromatography (HPLC) with UV detection. The methods were determined to be linear, precise and accurate (RSDs were lower than 5%), which allowed the quantitation of beta-blockers assayed at concentrations in the range 25-0.78 micrograms ml-1. After validation of reversed-phase HPLC methods, their analytical error functions were established by a rapid, simple and economical procedure. The discrimination of the best function for each active principle was performed by an appropriate polynomial statistical analysis, yielding SD (microgram ml-1) = 0.0295 + 0.0124C - 3.88 x 10(-4)C2 for celiprolol, 0.0199 + 0.011C - 1.27 x 10(-5)C3 for bisoprolol; and 0.0183 + 0.0089C - 9.68 x 10(-6)C3 for oxprenolol. These analytical error functions are an alternative to the weighting methods used in parameter estimation of beta-blockers. PMID- 8729639 TI - The development and performance of a radioimmunoassay for the analysis of ZM 213,689, the major metabolite of meropenem--a carbapenem antibiotic--in plasma and urine. AB - The development of a radioimmunoassay for the analysis of ZM 213,689, the major metabolite of meropenem found in the plasma and urine of rat, dog and humans, is described. The assay is rapid in order to minimise the effect of degradation of meropenem to ZM 213,689 in biological samples and has a working range of 0.08-3.5 mg l-1 (RSD < or = 15%). The antibody was specific for ZM 213,689 with cross reactivity to meropenem of only 0.4%. The synthesis of the immunogen and radiotracer involved a novel approach due to the multifunctional nature of ZM 213,689. PMID- 8729640 TI - An investigation of the properties of a "shielded" phase for the solid phase extraction of acidic and basic compounds. AB - The extraction properties of Hisep, a novel bonded phase for solid phase extraction, have been investigated using a number of acidic and basic analytes and compared with those of a conventional ODS bonded material. Marked differences in extraction properties for the test analytes propranolol, anisic acid and ICI 128436 from aqueous buffer were noted between the two materials. Thus, extraction onto the ODS material was by both ionic and reversed-phase mechanisms whilst retention on the Hisep phase appeared to be mediated predominantly by reversed phase hydrophobic interactions. The presence of dog plasma had no effect on the extraction of propranolol but did reduce the extraction efficiencies observed for anisic acid and ICI 128436 in a pH dependent manner. PMID- 8729641 TI - Quantitation of 5-hydroxymethylfurfural (5-HMF) and related substances in dextrose injections containing drugs and bisulfite. AB - The spectrophotometric test for 5-hydroxymethylfurfural and related substances, which is included in multiple compendial monographs for dextrose-containing injections, was modified to eliminate interference of bisulfite anion. Experimental formulations containing heparin or dopamine hydrochloride were used in this study. The modified assay was shown to be accurate, precise and rugged. With an appropriate standard, the method can be used to measure 5-HMF in solutions with an approximate quantitation limit of 0.06 ppm. PMID- 8729642 TI - Application of capillary zone electrophoresis for analyzing biotin in pharmaceutical formulations--a comparative study. AB - The detection limit and reproducibility of capillary zone electrophoresis (CZE) measurement of biotin were compared to those of a spectrophotometric method and those of the determination of the sulphur content by combustion of the biotin sample followed by coulometric titration of the formed SO2. Drug analysis showed that all three methods gave consistent results and were suitable for the determination of biotin. CZE was found to be the best method for the determination of pharmaceutical formulations containing biotin because of its high separation efficiency, short analysis time, ease of instrumentation and sample preconditioning, and good precision. PMID- 8729643 TI - A capillary gas-liquid chromatographic method for the assay of the neuroleptic drug zotepine in human serum or plasma. AB - A capillary gas-liquid chromatographic method suitable for the assay of the atypical neuroleptic drug zotepine in human serum or plasma was developed. A liquid-liquid extraction with three subsequent extraction steps was applied for sample preparation. The minimum detectable concentration was 1.0 ng ml-1. The within-day relative standard deviation (RSD) (n = 6) was 5.3% at 5 ng ml-1, 3.6% at 10 ng ml-1 and 6.1% at 100 ng ml-1. The day-to-day RSD (n = 6) was 9.3% at 10 ng ml-1 and 5.1% at 100 ng ml-1. Steady-state serum levels of four schizophrenic patients were measured. PMID- 8729644 TI - Development of a liquid chromatographic method for the control of related substances in chlorprothixene hydrochloride. AB - The development of a reversed-phase liquid chromatographic method, using a mobile phase containing a mixture of anion and cation ion-pairing agents and a base deactivated octyldecylsilyl column as stationary phase, is described for the control of all known impurities in (Z)-chlorprothixene hydrochloride (bulk drug). Validation of the method showed it to be reproducible, selective for both (Z) chlorprothixene hydrochloride and its E-isomer, accurate and linear over the concentration range of analysis with a limit of detection of 0.3 microgram ml-1. PMID- 8729645 TI - Direct chiral resolution of 15-deoxyspergualin using a cellobiohydrolase liquid chromatographic column. AB - An LC method for the direct determination of the enantiomers of 15 deoxyspergualin was developed, optimized and validated. A commercially available LC column containing a cellulase enzyme (cellobiohydrolase I) stationary phase was used. The effect of various parameters on the separation and validation data are discussed. PMID- 8729646 TI - Simultaneous quantitation of pioglitazone and its metabolites in human serum by liquid chromatography and solid phase extraction. AB - A high-performance liquid chromatographic (HPLC) method for the simultaneous determination of pioglitazone (U-72107) and its potential metabolites (M-1 to M 6) in human serum was developed. The method involved a solid phase extraction (SPE) of pioglitazone, its metabolites, and the internal standard (U-92573) from serum using C18 SPE columns with an elution solvent of 0.5 ml of acetonitrile water (35:65, v/v). Separation of the eight analytes was achieved within 20 min using a reversed-phase Zorbax RX-C8 analytical column (250 mm x 4.6 mm i.d., 5 microns particle size) with a mobile phase of acetonitrile-water (40:60, v/v) containing 3 ml acetic acid per liter mobile phase (apparent pH 5.5). An ultraviolet detector operated at 269 nm was used with a linear response observed from 0.02 to 2 micrograms ml-1 for these analytes except for M-4 which was best fitted with a polynomial regression. Limit of quantitation was found to be 0.02 microgram ml-1 for pioglitazone, M-3, M-5, and M-6; 0.04 microgram ml-1 for M-2 and M-4; and 0.5 microgram ml-1 for M-1 when using a 0.5 ml serum sample for extraction. Obtained from the method validation, intra- and inter-assay precision was < or = 9% and accuracy ranged from -8.2 to 13.4% for all analytes. The applicability of this method has been demonstrated by successfully analyzing clinical serum samples. The strategies in the HPLC characterization and in the SPE procedure development for this method are discussed as well. PMID- 8729647 TI - Comparison of two extraction methods for determination of propranolol and furosemide in human plasma by mixed-mode chromatography. AB - An isocratic high performance liquid chromatographic method is described for the determination of the beta-adrenergic blocking drug, propranolol, and the diuretic, furosemide, in human plasma. The two compounds and the internal standard were extracted from plasma using a two-step extraction technique. Propranolol and pindolol (internal standard) were first extracted from alkaline plasma into diethyl ether; this was followed by extraction of furosemide into acidified ether: hexane (65:35). The two extracts were then combined and evaporated under nitrogen, and the reconstituted residues were analysed on a C18/SCX reversed-phase/cation exchange column with a mobile phase of acetonitrile: 0.1 M sodium acetate pH 4 (33:67). The drugs and the internal standard were detected by UV absorption at 230 nm. The drugs were also extracted from plasma by a column-switching technique utilizing a ten-port valve. The drug compounds were retained on a C18 pre-column. A comparison of RSD for within-batch (intra-assay) and between-batch (inter-assay) runs for both methods was carried out, the liquid/liquid extraction method giving better recovery values. The calibration graphs were linear from 25-300 ng ml-1 for furosemide and 50-400 ng ml-1 for propranolol. Recovery values were > 90.0% by liquid/liquid extraction and > 76.0% by column switching. PMID- 8729648 TI - Determination of the polyfructosan sinistrin in biological fluids by HPLC with electrochemical detection. AB - A sensitive HPLC method with electrochemical detection was developed for the determination of the polyfructosan sinistrin in human plasma and urine. Proteins and interfering components such as glucose were removed from plasma and urine samples by solid phase extraction on C18 cartridges. Chromatographic separations were achieved at 85 degrees C on a 300 mm x 7.8 mm i.d. column, using ion moderated partition chromatography with distilled water at a flow rate of 0.6 ml min-1. After post-column addition of NaOH 0.3 M (0.6 ml min-1), the electrochemical detection of the eluate was performed with a sequence of three potentials (0.05 V, -0.8 V, 0.6 V) of specific pulse duration 300, 100 and 100 ms respectively. Xylose was used as internal standard for the quantitative determinations. The calibration curves were linear (r2 > 0.992) over the working range 5-300 micrograms ml-1. This method has been characterized, validated and applied successfully in a study comparing two modes of glomerular filtration rate determination in healthy volunteers (bolus vs. constant rate infusion of sinistrin). PMID- 8729649 TI - Quantification of zidovudine and individual zidovudine phosphates in peripheral blood mononuclear cells by a combined isocratic high performance liquid chromatography radioimmunoassay method. AB - Zidovudine (ZDV, AZT) is the first clinically effective drug licensed for use in the treatment of human immunodeficiency virus (HIV) infection. Activation of ZDV requires phosphorylation to ZDV triphosphate by cellular kinases. It is important, therefore, to determine the intracellular levels of the active form because measurement of ZDV concentrations in plasma have not reflected any direct relationship with activity or toxicity. In this paper a validated assay for the measurement of both ZDV and its three phosphorylated anabolites, ZDV mono-, di- and triphosphate, in peripheral blood mononuclear cells (PBMCs) is described. The method consisted of a combination of isocratic high performance liquid chromatography (HPLC) separation and radioimmunoassay (RIA). The PBMCs were separated from whole blood and ZDV and ZDV nucleotides were extracted and separated by isocratic elution with an ion-pairing mobile phase on a reversed phase HPLC column. The collected ZDV and individual ZDV nucleotide fractions were dephosphorylated to ZDV, cleaned by solid phase extraction and assayed by a commercially available RIA kit. The assay developed was successfully used to determine intracellular ZDV and anabolite concentrations of 10 PBMC samples taken from HIV positive patients on ZDV treatment. PMID- 8729650 TI - Determination of 1-benzo[b]thien-2-ylethanone and related impurities by high performance liquid chromatography. AB - 1-Benzo[b]thien-2-ylethanone (2-acetylbenzothiophene, 2-ABT) and related impurities were determined using a reverse-phase high performance liquid chromatography system and UV detection at 254 nm. Separation was achieved isocratically on a 4.6 mm x 25 cm, 5 microns Zorbax Rx-C8 column using an eluent which is 0.2% perchloric acid/THF in a ratio of 60:40 (v/v). The chromatographic system resolved 2-ABT and known impurities in less than 45 min with near baseline resolution. Known impurities were quantitated versus 2-ABT with corrections made for differences in detector response at the specified wavelength. Linearity for 2 ABT was demonstrated with a correlation coefficient > 0.9999. Assay precision (RSD values) for impurities at 0.5% ranged from +/- 1.8% to +/- 14%, while precision (RSD values) for the 2-ABT determination ranged from +/- 0.81% to +/- 1.1%. A variety of different chromatographic columns and conditions are discussed for the application. PMID- 8729651 TI - Determination of salmeterol in metered-dose and dry-powder inhalers by reversed phase high performance liquid chromatography. PMID- 8729652 TI - Oriented immobilization of antibodies and its applications in immunoassays and immunosensors. AB - The immobilization of antibodies on solid-phase materials has been used in many areas such as purification, diagnostic immunoassays and immunosensors. Problems associated with the loss of biological activity of the antibodies upon immobilization have been noticed in many cases. One of the main reasons for such loss is attributed to the random orientation of the asymmetric macromolecules on support surfaces. In this paper, the approaches for achieving oriented coupling of antibodies to increase the antigen binding capacity are reviewed. Some issues such as steric hindrance caused by neighbouring antibody molecules, the distance between an antibody and the support surface and the use of antibody fragments are dealt with. Some applications of the oriented immobilized antibodies in immunoassays and immunosensors are examined. PMID- 8729653 TI - Multi-element, multi-media method for the determination of airborne elemental emissions by inductively coupled plasma atomic emission spectrometry. AB - The exposure of people to airborne emissions of toxic elements, particularly lead, is well documented. The approach in most studies has been to examine the concentrations of a single element in the air or in the blood of the local population. In urban industrialized settings there can often be many sources of a particular element and the element may not be released alone. In order to establish a full risk assessment and determine the source of emissions, it is important to determine all toxic elements in a variety of media. This paper presents a single analytical procedure for the determination of 16 toxic elements in solids, soils, wipes and glass-fibre filters. The procedure includes adapting an acid digestion method (USEPA SW 846 Method 3050B) and analysis of the digestate by inductively coupled plasma atomic emission spectrometry (USEPA SW 846 Method 6010B). All 16 elements were determined in this medium over several orders of magnitude. Accurate and precise results were obtained to less than 5 ng m-3. The procedure compared well with established procedures for lead. PMID- 8729654 TI - Gas chromatographic determination and negative-ion chemical ionization mass spectrometric confirmation of 4,4'-methylenebis(2-chloroaniline) in urine via thin-layer chromatographic separation. AB - An analytical method for monitoring 4,4'-methylenebis(2-chloroaniline) (MOCA) in urine was developed. The MOCA extract from a urine sample was derivatized with pentafluoropropionic anhydride and subsequently cleaned-up via TLC before being analysed by a gas chromatograph equipped with an electron capture detector. Heptafluorobutyric anhydride derivatized MOCA was used as the internal standard during quantification. The method showed that the analytical interferences are effectively minimized by using the TLC technique for sample clean-up. Recoveries of spiked MOCA in urine at levels of 7.5, 22 and 62.5 micrograms dm-3 were all over 90% with the limit of detection determined to be 1 microgram dm-3. Confirmation of MOCA in a urine sample was performed by using a gas chromatograph mass spectrometer in negative-ion chemical ionization mode. PMID- 8729655 TI - Development and evaluation of a chemiluminescent immunoassay for chlortoluron using a camera luminometer. AB - The feasibility of using an enhanced chemiluminescent immunoassay for screening water samples was adapted to a Dynatech Microlite camera luminometer and assessed to provide a semiquantitative assay based on a photographic record of the luminescent end-point. An enhanced chemiluminescent immunoassay of the herbicide chlortoluron was chosen in this instance as an example, although other pesticide compounds could equally well have been used provided that suitable antisera were available. This luminescent assay has shown considerable potential by providing a rapid, simple and portable means of monitoring multiple water samples for the presence of chlortoluron. The assay was able to identify samples containing the herbicide at or above the European limit for individual pesticides in drinking water [the EU Maximum Admissible Concentration (MAC) = 0.1 microgram l-1]. A 100% accuracy was obtained in the analysis of samples containing chlortoluron at concentrations above the range of 0.07-0.12 microgram l-1; however, for concentrations within this range the results were 65% accurate. Consequently, the method could greatly facilitate increased monitoring for the presence of chlortoluron in water supplies and in other areas of environmental analysis. PMID- 8729656 TI - Microtitre-plate enzyme-linked ligand-sorbent assay of riboflavin (vitamin B2) in human plasma and urine. AB - Enzyme-linked ligand-sorbent assay (ELLSA) of riboflavin was performed in standard, multi-well microtitre plates. 3-Carboxymethylriboflavin was carbodiimide-coupled to bovine serum albumin and the conjugate was adsorbed on the well surface. Riboflavin-binding protein from egg-white was biotinylated with biotinamidocaproate N-hydroxysuccinimide ester. The assay was based on competition of riboflavin analyte with the immobilized flavin for the biotinylated binder. Secondary adsorption of the biotinylated riboflavin-binding protein was measured by using avidin-bearing horseradish peroxidase label. The optimized method had a detection limit of 0.8 pmol of riboflavin and was expected to work within a riboflavin concentration range of 2 X 10(-8)(-4) X 10(-6) mol l 1. Preliminary trials suggested that ELLSA was suitable for determining riboflavin in human urine and the sum of riboflavin and flavin nucleotides in human plasma. The analytical performance of ELLSA for those materials was characterized by good consistency of the results with those obtained by conventional, fluorimetric methods, a mean recovery of riboflavin supplement of over 90% and a within-plate relative standard deviation below 20%. Some unique samples of both urine and plasma were assayed with between-plate relative standard deviations higher than 30%, implicating further modification of this version of ELLSA. The method intended for routine control determinations of vitamin B2 status in human subjects and is addressed to laboratories that routinely perform automated, microplate-based enzyme-linked assays. PMID- 8729657 TI - Determination of trace levels of niguldipine in urine and blood by adsorptive stripping voltammetry at the hanging mercury drop electrode. AB - A relatively simple electroanalytical procedure for the determination of niguldipine in biological samples is described. The technique involves adsorptive accumulation of the drug at the hanging mercury drop electrode (HMDE) followed by a differential-pulse polarographic determination of the preconcentrated species. The adsorptive stripping response is evaluated with respect to various experimental conditions, such as solvent composition and pH of the supporting electrolyte, accumulation potential and accumulation time. After a simple sample preparation, the method can be used for the determination of niguldipine in blood and urine. Interfering substances are simply removed by precipitation, adding a small amount of 5% ZnSO4 solution and ethanol to the urine or blood sample and centrifuging the mixture. A limit of detection of 6.7 ng per ml of urine and 41 ng per ml of blood is found with a mean recovery of 96% in urine and 71% in blood. The mean relative errors are 8.4% and 2.2%, respectively. PMID- 8729658 TI - Cow milk allergy in infancy and early childhood. PMID- 8729659 TI - Is formaldehyde an important cause of allergic respiratory disease? PMID- 8729660 TI - Chloride transport and the action of sodium cromoglycate and nedocromil sodium in asthma. PMID- 8729661 TI - Distinct patterns of cow's milk allergy in infancy defined by prolonged, two stage double-blind, placebo-controlled food challenges. AB - BACKGROUND: The clinical manifestations of cow's milk allergy (CMA) are highly variable, and challenges usually identify only immediate, IgE mediated reactions. OBJECTIVE: To clearly identify CMA of immediate and delayed types using a two stage, double-blind, placebo-controlled food challenge (DBPCFC), and to prospectively compare the clinical history and analyses of specific IgE antibodies to milk in predicting outcome of DBPCFC. METHODS: A total of 69 patients (33 girls, 36 boys) were recruited for study based on a history highly suggestive of CMA and resolution of symptoms on a bovine protein-free diet. After skin-prick tests (SPTs) and search for allergen-specific serum IgE antibodies by enzyme allergosorbent test (EAST), a two-stage DBPCFC was performed over several days. RESULTS: Of 16 patients (mean age 36.9 months) classified as probable immediate reactors based on the history, 10 (62.5%) had a positive DBPCFC with similar patterns to historical adverse reactions (< or = 2 h after milk exposure). The other 53 (77%) patients (17.3 months) had a history of probable delayed type CMA presenting with predominantly gastrointestimal symptoms from 2 h and up to 6 days after milk exposure. Of these, 15 (28.8%) had a positive DBPCFC, again with a symptom pattern similar to the history. Sensitivity/specificity of SPT was similar to that of EAST for both immediate (70/83% and 62/83% respectively, NS) or delayed (0/97% and 0/97%) CMA confirmed by DBPCFC. CONCLUSIONS: Using our two-stage, prolonged DBPCFC, we clearly identified two groups of children with CMA, reflecting different pathogenesis of either immediate-type IgE-dependent, or delayed-type IgE-independent allergy. Although useful in immediate reactors, IgE antibody determination cannot predict the outcome of DBPCFC in delayed reactors. A thorough clinical history was the most helpful tool to predict the type of response in challenge positive patients. PMID- 8729662 TI - Does an allergy skin test on school-children predict respiratory symptoms in adulthood? AB - BACKGROUND: It is interesting from the correct point of preventive health care whether allergy skin tests at entry have the positive relationship with common respiratory symptoms such as persistent cough, persistent phlegm and wheeze with colds at end point in longitudinal studies. OBJECTIVE: The purpose of this study is to investigate this relationship in subjects followed from when they were school-children until they were young adults. METHODS: Young adults aged 18-31 years who had participated in an allergy skin test and a health survey for common respiratory symptoms during primary and junior high school were asked about their respiratory symptoms at the end-point in the follow-up study. Three cohort groups were employed in the follow-up study. RESULTS: According to a multiple logistic analysis with adjustments made for sex, symptoms at entry in childhood, and age and smoking habits at the end-point, young adults with a positive skin test to an extract of house dust as school-children showed a higher prevalence of wheeze with colds compared with those with a negative skin test. However, this relationship was not found for persistent cough or persistent phlegm, indeed, the latter symptom was significantly associated with negative skin tests in one cohort group. CONCLUSION: The respective symptoms at entry in childhood were also predictors of their occurrence in young adults. The results show that the common respiratory symptoms such as persistent phlegm and wheeze with colds in childhood are related to those in young adults rather than is an allergy skin test to an extract of house dust in childhood. PMID- 8729663 TI - Pulmonary surfactant given prophylactically alleviates an asthma attack in guinea pigs. AB - BACKGROUND: Previous studies have indicated that the increased airway resistance that develops in asthma may partly be due to a surfactant dysfunction. If so, it might be possible to alleviate the acute signs following an allergen challenge by prophylactically instilling into the airways a well functioning pulmonary surfactant. OBJECTIVE: The study was planned and enacted to test the above hypothesis. METHODS: The lung function (airway resistance, tidal volume, minute ventilation, and dynamic compliance) of 22 immunized guinea-pigs was studied for 30 min following a challenge. Ten of the animals had received a tracheal instillation of 0.5 mL calf lung surfactant extract (CLSE, 35 mg/mL) prior to the challenge. RESULTS: The animals receiving the dose of 17.5 mg surfactant were less affected by the challenge than were the controls. Only one of them died following the challenge, whereas four of the 12 controls succumbed. Lung function was significantly less affected among the nine surviving animals treated with surfactant prior to the challenge than among the eight surviving controls (P < 0.01) and also their blood gases (pCO2 and pO2) were less influenced (P < 0.05). CONCLUSION: The study indicated that the symptoms developing after a challenge, which to some extent simulate those of asthma, can be alleviated by a prophylactic airway instillation of pulmonary surfactant. PMID- 8729664 TI - Exposure to gaseous formaldehyde induces IgE-mediated sensitization to formaldehyde in school-children. AB - BACKGROUND: Children attending a primary school showed symptoms such as headache, cough, rhinitis and epistaxis. Assessment of specific IgE to formaldehyde gave positive results in some children. OBJECTIVE: Was IgE-mediated sensitization as well as symptoms in children associated with formaldehyde exposure at school? METHODS: Sixty-two 8-year-old children attending three forms at a primary school were investigated. Indoor formaldehyde concentrations were measured in classrooms of both schools (one frame construction with particleboard used extensively as panelling vs a brick building) which were consecutively attended. Assessment of specific IgE to formaldehyde was done in all children. Children were transferred to a brick building and 3 months later specific IgE to formaldehyde in pupils showing initially elevated radioallergosorbent test (RAST) values reassessed. In all children symptoms were evaluated by questionnaire before and 3 months after changing school. RESULTS: In the school panelled with particleboard the World Health Organization (WHO) threshold for formaldehyde of 0.050 ppm was crossed in two classrooms (0.075 ppm and 0.069 ppm) whereas in one classroom 0.043 ppm was found. RAST classes of > or = 2 were found in three children, two of them attending the classroom with 0.075 ppm formaldehyde. Elevated RAST classes of > or = 1.3 were found in another 21 pupils. Thirty-eight pupils as well as 19 control children showed RAST classes in the normal range of < or = 1.2. Headache, nose bleeding, rhinitis, fatigue, cough, dry nasal mucosa and burning eyes were found in the affected children. There was a good correlation between symptoms and the formaldehyde concentrations in the classrooms. However, elevated IgE levels to formaldehyde did not correlate with symptoms. Formaldehyde concentrations in the classrooms of the brick built school were 0.029 ppm, 0.023 ppm and 0.026 ppm. After transferral specific IgE to formaldehyde decreased significantly from 1.7 +/- 0.5 to 1.2 +/- 0.2 (P < 0.002) as did the incidence of symptoms. CONCLUSION: Gaseous formaldehyde, besides its irritant action, leads to IgE-mediated sensitization. As children are more sensitive to toxic substances than adults, threshold levels for indoor formaldehyde should be reduced for children. PMID- 8729665 TI - Recovery of human basophils after FMLP-stimulated secretion. AB - BACKGROUND: Basophils are circulating, secretory granulocytes that are generally considered to be end-stage cells. In one species of guinea-pigs, basophilic leucocytes have been shown to recover from stimulated secretion in short-term cultures. Similar studies have not been done using human basophils. OBJECTIVE: The purpose of this study was to examine human basophils in short-term recovery intervals following stimulation of secretion to determine whether visual evidence of recovery occurred. METHODS: We examined the ultrastructural morphology of early recovery (10 min-6 h) of human basophils following secretion stimulated by formyl-methionyl-leucyl-phenyl-alanine (FMLP). A combined technique for electron microscopy consisted of post-fixation exposure to cationized ferritin and reduced osmium, providing maximum quality images and allowing identification of intracellular spaces/organelles that opened to the cell surface, often out of the plane of section. RESULTS: The ultrastructural evaluation revealed that control basophils (0 time-6 h) did not undergo regulated secretion or develop the morphologies associated with recovery following secretion. FMLP-stimulated basophils underwent an overlapping continuum of piecemeal degranulation- >anaphylactic degranulation (0 time-1 min), producing vesicle- and granule-free, completely degranulated, viable, mature basophils with polylobed nuclei. The early recovery period (10 min-6 h) following FMLP stimulation was characterized by reconstitution of granules. Morphological mechanisms for granule reconstitution included a mixture of conservation, condensation, and synthetic events. CONCLUSION: Human basophils, like guinea pig basophils, have the potential to recover from regulated secretion. PMID- 8729666 TI - Diminution of T cells with gamma delta receptor in the peripheral blood of allergic asthmatic individuals. AB - BACKGROUND: T cells are shown to regulate allergy and asthma. They are heterogeneous by virtue of surface receptor subtypes (alpha beta and gamma delta receptors), however, their pathophysiological roles in asthma are unclear. OBJECTIVE AND METHODS: The present study employed flow cytometric analysis to compare the size of T-cell subsets and eosinophils present in the peripheral blood of healthy, allergic and allergic asthmatic individuals. RESULTS: Current results demonstrated that the percentages of gamma delta T cells declined from 4.1% in healthy to 3.2% in the allergic subjects and to a significantly lower (P = 0.01) 2.4% in allergic asthmatics. The absolute numbers of circulating gamma delta T cells also were diminished in a similar fashion such that healthy individuals had a significantly higher mean cell count (91.8 x 10(3)/mL) than did allergic asthmatics (47.8 x 10(3)/mL) (P = 0.0266). In contrast, alpha beta T cells were comparable in the healthy, allergic, and allergic asthmatic populations (65.3%, 65.8% and 69.4%, respectively); the differences were not statistically significant. On a populational basis, the proportion of individuals having both gamma delta T cells < or = 4.1% and eosinophils > or = 2.1% was the lowest in the healthy population (30.8%), but was elevated in the allergic group (85.7%) as well as in the allergic asthmatic group (86.6%). However, on an individual basis, those who had reduced gamma delta T cells did not have consistently higher eosinophil counts or IgE level. CONCLUSION: It was concluded that no significant correlation existed between the levels between gamma delta T cells and eosinophils or between gamma delta T cells and IgE present in the peripheral blood. This report, for the first time, documents that allergic asthmatics have reduced gamma delta T cells with reciprocally elevated eosinophil numbers in their peripheral circulation. However, it does not indicate that the reduction of gamma delta T cells directly correlates with the predominance of eosinophils of IgE levels in the diseased populations. The pathophysiological role of gamma delta T cells in allergic diseases awaits further exploration. PMID- 8729667 TI - Immunoglobulin E in acute phase response to surgical stress. AB - BACKGROUND: A few years ago we reported that acute myocardial infarction is associated with a distinct, transient rise in serum immunoglobulin (Ig)E. OBJECTIVE: We wondered whether this response is specific only for myocardial infarction or whether it reflects a more generalized phenomenon, precipitated by tissue injury. METHODS: We carried out a large prospective study on 149 patients undergoing various surgical procedures. These were the patients undergoing coronary artery bypass surgery, who did (n = 39) or did not (n = 42) develop perioperative myocardial infarction, patients subjected to various thoracic operations (n = 33), cholecystectomy (n = 17) or repair of the inguinal hernia (n = 18). Thirty healthy volunteers formed the control group. RESULTS: In all groups studied, except the control group, serum IgE levels began to rise shortly after the operation, reached a peak by the fifth postoperative day, and then gradually declined. This was in striking contrast to serum immunoglobulin G which soon after the operation became markedly depressed, and took more than a week to return to preoperative values. Specific IgE against common aeroallergens, measured in a subgroup of patients with initially high total serum IgE, showed no uniform pattern of change. CONCLUSION: Surgical operations are accompanied by a characteristic, transient rise in serum IgE concentration. Immunoglobulin E might be involved in acute phase response to tissue injury. PMID- 8729668 TI - IgE and monoclonal antibody binding by the mite allergen Der p 7. AB - BACKGROUND: The recently characterized group 7 house dust mite allergens give positive skin-test reactions in 53% of allergic patients. OBJECTIVE: This study was performed to compare the IgE binding activity of natural and recombinant Der p 7, to measure the binding in allergic sera in comparison to major allergen Der p 2 and characterize the response by competitive inhibition with monoclonal antibodies. METHODS: IgE anti Der p 2 and Der p 7 antibodies against the recombinant allergens and monoclonal binding activities were measured by a solid phase radioimmune assay. RESULTS: A competitive binding assay showed that rDer p 7 inhibited 91% of IgE-binding to natural Der p 7 in 2 sera and 73% in a further two. The IgE binding of rDer p 2 and Der p 7 from 41 sera was then compared. Of the sera 88% and 46% respectively showed positive binding. All of the 19 sera which bound Der p 7 also bound Der p 2 but 11 (58%) had bound IgE to Der p 7 as high or higher than the binding to Der p 2. These sera were mostly high responders to both allergens. A panel of six monoclonal antibodies produced against either rDer p 7 or rDer f7 was used for epitope analysis. All of these reacted with each allergen by enzyme linked immunosorbent assay (ELISA) and immunoblotting. Two patterns of cross inhibition of monoclonal antibody binding were observed and of five monoclonal antibodies tested, four could inhibit the binding of IgE (WH9, WH22, WP8 and HD19) while one (WH14) could not. CONCLUSIONS: Although Der p 7 only reacts with 50% of allergic sera it often has a high IgE binding activity and may be more important than the major Der p 2 allergen in a high percentage of subjects. The combined competitive inhibition experiments show the IgE response is directed at several specificities. PMID- 8729669 TI - Allergy to olive pollen: T-cell response from olive allergic patients is restricted by DR7-DQ2 antigens. AB - BACKGROUND: We have recently described the association between the IgE antibody response to Ole e I (the major antigen from olive tree pollen) and the DR7-DQ2 haplotype in a Spanish population. OBJECTIVE AND METHODS: Due to the linkage disequilibrium between DR7 and DQ2, and thus the difficult distinction between the role of these two antigens in the T-cell activation response, we decided to solve this question by two approaches: 1. The study of another ethnic group, individuals of Arabic origin, with a presumably distinct disequilibrium linkage between DR and DQ antigens. Genomic DNA typing was performed in 46 subjects (allergic and non-allergic) by Restriction Fragment Length Polymorphism (RFLP) and results showed that patients with specific IgE antibodies alpha-Ole e I, were DR7 and/or DQ2. These data show a similar restriction pattern to those previously described for Spanish patients. The phenotypic frequency of DR7 antigen is significantly greater than in the non-allergic population, with a corrected P (Pc) value of 0.03. 2. The analysis of the genetic requirements of Ole e I response, using T-cell lines specific for this antigen. This was first carried out by blocking the proliferative response of these T-cell lines with specific anti-human HLA class II antibodies and then testing the genetic restriction of this response using a panel of histocompatible and histoincompatible Antigen Presenting Cells (APCs). Both experiments corroborate the hypothesis that DR7 and DQ2 are implicated in the recognition of Ole e I. PMID- 8729670 TI - The importance of the pecan tree pollen in allergic manifestations. AB - BACKGROUND: Pecan tree pollen is considered to be highly allergenic. However, no specific scientific data about its role in causing allergic diseases are available. OBJECTIVE: To study the role of pecan tree pollen in the development of allergy. METHODS: The presence of pecan tree pollen was determined by weekly and monthly counting of airborne grains. The incidence of pecan tree pollen atopy and clinical manifestations were studied in 395 participants, aged 4-70 years, who comprised 78.2% of the whole eligible population of a rural community. The participants were skin tested for different extracts of allergens, completed detailed questionnaires, and their medical files were evaluated. RESULTS: During May, pecan tree pollen grains comprised 70% of the total airborne grains. A positive skin-prick test (SPT) to pecan was shown by 46 (11.6%) participants, constituting 25.4% of the atopic population. Of those who were found atopic to one or more allergens 50.3% had symptoms, whereas the parallel figure for those atopic to pecan pollen was 76.1% (P < 0.005); 58.7% of the pecan atopic participants had hay fever, 43.5% had asthma, and 31.5% had both hay fever and asthma. Among pecan atopic participants the incidence of hay fever increased with age (P = 0.05), while the incidence of bronchial asthma, as a sole manifestation of allergy, decreased in the > 17-year-old age group (P < 0.01). Of the pecan atopics 65.2% had clinical symptoms coinciding only with the pecan pollen season and an additional 10.9% had perennial symptoms. CONCLUSION: Pecan tree releases highly allergenic pollen grains, which are correlated to the incidence of hay fever in the exposed population. The contribution of pecan tree pollen to the symptoms was highly significant after discounting olive and cypress trees that also pollinate in the spring. In children, the pecan tree constitutes a possible etiologic agent for the development of asthma. PMID- 8729671 TI - Inhalant allergy to fresh asparagus. AB - BACKGROUND: Two patients experienced itching conjunctivitis, running nose, tightness of the throat and coughing during preparation of fresh asparagus. Eating asparagus after cooking did not provoke any allergic symptoms. Both patients were atopic, sensitized additionally to pollens of grasses and trees as well as to onion. OBJECTIVE: To assess the hypersensitivity reactions to fresh and heated asparagus and to investigate any crossreactivities among the allergens. METHODS: Skin-prick tests were performed with commercial allergens and native asparagus and the patients were tested with Pharmacia CAP system for specific IgE antibodies against asparagus, onion, garlic, birch pollen, mugwort pollen and two recombinant birch pollen allergens, Bet v I and profilin. Inhibition of IgE antibody binding to solid phase homologous and unrelated allergens by increasing doses of liquid allergens (inhibitors) was studied. RESULTS: Skin-prick tests with native green and white asparagus were strongly positive, but negative with cooked asparagus. Both patients had measurable levels of IgE antibodies against asparagus (3.0 and 6.2 kU/L respectively) and several other allergens. One patient was highly sensitive to birch and Bet v I. Both were positive to profilin, mugwort and onion. In all cases the antibody uptake could be extensively and specifically inhibited by homologous allergen. The asparagus specific IgE antibodies of the two patients could only be inhibited by asparagus. No inhibition was obtained after heating of the asparagus extract to 100 degrees C. CONCLUSIONS: The patients were specifically sensitized by asparagus. No immunological crossreactions could be observed. The measurements of IgE antibodies to other allergens were also specific, representing parallel multiple sensitivity. Profilin inhibited profilin-specific IgE binding but did not react with the asparagus-specific IgE antibodies of these patients. The asparagus allergen recognized by the specific IgE antibodies of these patients was thermolabile. PMID- 8729672 TI - Anaphylaxis to penicillins after non-therapeutic exposure: an immunological investigation. AB - BACKGROUND: There are instances where individuals may come into contact with penicillins without being aware of it. This non-therapeutic exposure from different sources may cause sensitization and even clinical manifestations in subjects allergic to penicillins. OBJECTIVE: To determine the capacity that inadvertent contact with penicillins may have in inducing resensitization and symptoms in patients diagnosed as allergic to penicillins who were followed over a long period of time after their initial evaluation. METHODS: A follow-up study of penicillin-allergic subjects who agreed to be regularly tested for in vitro and in vivo control of their sensitivity. Skin tests were carried out with major and minor determinants of benzylpenicillin (BPO and MDM), amoxicillin (AX), and ampicillin (AMP), and specific IgE antibodies were determined by radioallergosorbent test (RAST). A questionnaire was sent to and answered by the subjects in order to see if they experienced symptoms at any time during the follow-up period. In addition, if any unexplained symptoms occurred, a bleeper system was used to contact the allergy centre. RESULTS: Seven subjects experienced anaphylactic reactions with no obvious cause. At the time of their initial allergic reaction, which was caused by exposure to prescribed penicillin, the subjects had one or more positive skin tests and/or RAST results to penicillin related reagents. However, over the following 2-4 years all their tests became negative. After reporting their unexplained reaction all seven had one or more positive skin tests and/or RAST results again and when retested 1 week later RAST measurements showed that levels of penicillin-specific IgE were maintained or increased. None of the subjects had knowingly received penicillin but the questionnaire showed that six had been exposed to it and in the seventh case exposure was likely. In two cases contact was by sexual intercourse with a partner who was receiving penicillin, three subjects had handled penicillin formulations and one had drunk from a glass previously used for giving penicillin. In the seventh case exposure could have occurred whilst in hospital for surgery, although this was not proven. CONCLUSIONS: these results show that non-therapeutic exposure to penicillin can cause severe symptoms and that in vitro and in vivo testing can help in the diagnosis of such cases. PMID- 8729673 TI - Crossreactivity between allergens in natural rubber latex and banana studied by immunoblot inhibition. AB - BACKGROUND: An association between allergic reactions to natural rubber latex and to banana has been reported but the immunochemical properties of the putative cross-reacting allergens remain unknown. OBJECTIVE: To study extracts of banana and natural rubber latex and sera from latex-allergic patients for possible crossreacting allergens and IgE antibodies. METHODS: Sera from 22 latex-allergic patients and 22 control subjects with no evidence of allergy to latex or to banana were studied. All patients had positive and controls negative reactions in skin-prick testing using an eluate of latex gloves. IgE antibodies to natural rubber latex and to banana were evaluated by immunoblotting and by radioallergosorbent test (RAST) and crossreactivity between allergens in banana and natural rubber latex by immunoblot inhibition. Skin-prick testing was used to examine in vivo reactivity to banana. RESULTS: Ten of the 22 (45%) latex-allergic patients sera recognized altogether 14 allergens in banana by immunoblotting. The most frequently identified banana allergens were 23, 32, 36, 39 and 47 kDa proteins. The banana skin-prick test was positive in 14 of 18 (78%) latex allergic patients studied and banana RAST in 12 of 14 patient sera tested. Fourteen of 21 interviewed patients reported symptoms from eating or handling bananas. In immunoblot inhibition studies a dose-dependent inhibition of IgE binding to banana extract with natural rubber latex proteins was observed in all five patient sera tested and, likewise, the binding of IgE to natural rubber latex extract was inhibited with banana proteins in four of the five patient sera. CONCLUSIONS: The present results confirm the existence of crossreacting allergens in natural rubber latex and banana and provide new information on the immunochemical nature and heterogeneity of these allergens. PMID- 8729674 TI - American cockroach Cr-PI allergen induces lymphocyte proliferation and cytokine production in atopic patients. AB - BACKGROUND: Previous studies have shown cockroach-induced antigen-specific IgE mediated asthma. In cockroach-infested areas, more then 50% of asthmatic subjects may have positive skin reactions to this allergen. Partial purified Cr-PI allergen from American cockroaches contains allergens with molecular weights of 72 and 78 kDa; however, little is known about its effect on the lymphocyte proliferation and cytokine production. OBJECTIVE: IgE synthesis is known to be regulated by interleukin-4 (IL-4) and interferon gamma (IFN gamma). Therefore, we studied Cr-PI allergen-induced cytokine production in atopic patients and healthy normal controls to understand each factors' role in the disease. METHODS: Peripheral blood mononuclear cells (PBMC) from cockroach skin-sensitive patients and controls were stimulated with mitogen and Cr-PI for proliferative response and cytokine production. Cr-PI antigen-specific T-cell cultures of atopic patients and healthy normal controls were used to test Cr-PI-induced proliferation and cytokine mRNA expression. RESULTS: PMBC of atopic subjects showed a significantly (P < 0.01) higher stimulation index for Cr-PI induced proliferation (SI = 11.8 +/- 3.7) when compared with that of non-atopic subjects (SI = 4.1 +/- 0.8) and cord bloods (SI = 2.1 +/- 0.4). Cr-PI-induced IL-4 was observed only in the PBMC of atopic patients, whereas Cr-PI-induced IFN gamma was detected in both atopic patients and normal controls. Likewise, Cr-PI-induced IL 4 mRNA expression in T-cell cultures was detected in all atopics but only one of nine controls. CONCLUSION: IL-4 mRNA expression and IL-4 production in PBMC and T cell cultures of atopic patients showed good correlation with clinical symptoms, skin-reactivity, specific IgE and proliferative response to Cr-PI. These results suggests that cockroach allergen may be a hidden cause of asthma and other atopic diseases. PMID- 8729675 TI - Sensory neuropeptides are not directly involved in bronchial hyperresponsiveness induced by interleukin-8 in guinea-pigs in vivo. AB - BACKGROUND: Interleukin-8 (IL-8) has been shown to be a chemotactic factor for neutrophils, T-lymphocytes and eosinophils. Repeated intranasal administration of IL-8 enhances bronchial responsiveness to inhaled histamine in guinea-pigs. Neuropeptides which are released from C-fibre nerve-endings have been postulated to induce bronchial hyperresponsiveness through neurogenic inflammation. OBJECTIVE: This study was conducted to examine whether sensory neuropeptides are involved in the IL-8-induced bronchial hyperresponsiveness. METHODS: IL-8 at a dose of 5 micrograms/kg was administered intranasally to guinea-pigs twice a week for 3 weeks. One day after the last administration, animals were anesthetized and artificially ventilated through tracheal cannula, and lateral pressure at the tracheal cannula (Pao) was measured as an overall index of airway responses to increasing concentrations of inhaled histamine (25, 50, 100, and 200 micrograms/mL). A NK1 and NK2 dual antagonist FK224 (10 mg/kg), a selective NK1 antagonist FK888 (10 mg/kg) or vehicle was intravenously administered 10 min before measurement of bronchial responsiveness. RESULTS: The IL-8 treatment significantly enhanced bronchial responsiveness to histamine (ANOVA P < 0.01). FK224 or FK888 did not alter the IL-8-induced bronchial hyperresponsiveness. CONCLUSION: We conclude that repeated intranasal administration of IL-8 causes bronchial hyperresponsiveness (BHR) and that neuropeptides such as neurokinin A and substance P do not directly contribute to the development of BHR induced by IL-8. PMID- 8729676 TI - Sodium channel slow inactivation and the distribution of sodium channels on skeletal muscle fibres enable the performance properties of different skeletal muscle fibre types. AB - Na+ currents (INa) and membrane capacitance were studied with the loose patch voltage clamp technique and action potential properties were studied with a two electrode voltage clamp on the end-plate, at the end-plate border and on extrajunctional membrane of skeletal muscle fibres. Slow inactivation regulates the available INa and is operative at the resting potential of both rat and human fibres. At the resting potential, slow inactivation causes a greater reduction in INa in fast-than in slow-twitch fibres. The relative resistance of slow-twitch fibres to slow inactivation may enable slow-twitch fibres to remain tonically active. Na+ channel inactivation may provide a peripheral mechanism that limits the duration that fast-twitch fibres can fire at high rates to prevent injury associated with prolonged high-frequency contraction. Consequently, slow inactivation may enable fast-twitch fibres to operate phasically at high rates or slow-twitch fibres to fire continuously at lower rates. For both fast- and slow twitch fibres. INa normalized to membrane area was greatest on the end-plate, intermediate on the end-plate border and smallest on extrajunctional membrane. When normalized to membrane capacitance. INa was the same on the end-plate and the end-plate border and smallest on extrajunctional membrane. For a given membrane region, INa was larger on fast- than on slow-twitch fibres. The higher density of Na+ channels near the end-plate increased the safety factor for neuromuscular transmission by lowering the action potential threshold and increasing the action potential rate of rise at the end-plate. PMID- 8729677 TI - Effect of intracellular and extracellular ion changes on E-C coupling and skeletal muscle fatigue. AB - The causative factors in muscle fatigue are multiple, and vary depending on the intensity and duration of the exercise, the fibre type composition of the muscle, and the individual's degree of fitness. Regardless of the aetiology, fatigue is characterized by the inability to maintain the required power output and the decline in power can be attributed to a reduced force and velocity. Following high-intensity exercise, peak force has been shown to recover biphasically with an initial rapid (2 min) recovery followed by a slower (50 min) return to the pre fatigued condition. The resting membrane potential depolarizes by 10-15 mV, while the action potential overshoot declines by a similar magnitude. Following high frequency stimulation of the frog semitendinous muscle, we observed intracellular potassium [K+]1 decrease from 142 +/- 5 to 97 +/- 8 mM, while sodium [Na+]i rose from 16 +/- 1 to 49 +/- 6 mM. The [K+]i loss was similar to that observed in fatigued mouse and human skeletal muscle, which suggests that there may be a limit to which [K+]i can decrease before the associated depolarization begins to limit the action potential frequency. Fibre depolarization to- 60 mV (a value observed in some cells) caused a significant reduction in the t-tubular charge movement, and the extent of the decline was inversely related to the concentration of extracellular Ca2+. A decrease in intracellular pH (pHi) to 6.0 was observed, and it has been suggested by some that low pH may disrupt E-C coupling by directly inhibiting the SR Ca2+ release channel. However, Lamb at al. (1992) observed that low pH had no effect on Ca2+ release, and we found low pHi to have no effect on t-tubular charge movement (Q) or the Q vs. Vm relationship. The Ca2+ released from the SR plays three important roles in the regulation of E C coupling. As Ca2+ rises, it binds to the inner surface of the t-tubular charge sensor to increase charge (Q gamma) and thus Ca2+ release, it opens SR Ca2+ channels that are not voltage-regulated, and as [Ca2+]i increases further it feeds back to close the same channels. The late stages of fatigue have been shown to be in part caused by a reduced SR Ca2+ release. The exact cause of the reduced release is unknown, but the mechanism appears to involve a direct inhibition of the SR Ca2+ channel. PMID- 8729678 TI - Fatigue vs. shortening-induced deactivation in striated muscle. AB - Fatigue and shortening-induced deactivation, two conditions that both lead to reversible depression of the mechanical performance of striated muscle are briefly reviewed. Fatigue. Isolated fibres from frog skeletal muscle (1-3 degrees C) that are stimulated to produce a 1 s fused tetanus at 15 s intervals are brought into a state of myofibrillar fatigue, (tetanic force reduced to 70-75% of the control) that is attributable to reduced performance of the myofibrils with no significant change in activation of the contractile system. A more intense stimulation programme (a single stimulus applied at 1-2 s intervals) reduces the tetanic force below 70% of the rested-state level. Under these conditions, failure of activation becomes increasingly important as a cause of the force decline. Deficient inward spread of activation is likely to account for at least part of the force decline after a period of intense fatiguing stimulation. Shortening-induced deactivation. Striated muscle that is allowed to shorten during activity loses some of its capacity to produce force, full restoration of the contractile strength being attained 1-2 s after the shortening phase. The depressant effect of shortening is demonstrable in skinned preparations as well as in intact muscle fibres and the magnitude of the effect is dependent on the state of activation of the muscle fibre when the movement occurs. The experimental evidence supports the view that sliding of the thick and thin filaments during activity reduces the affinity for calcium at the regulatory sites on the thin filament, leading to a transitory deactivation of the contractile system. PMID- 8729679 TI - Slow recovery of force in single skeletal muscle fibres. AB - After a bout of intense exercise, especially in untrained persons recovery of muscle force is often slow. Force depression is much more marked at low frequencies of stimulation than at high frequencies ("low-frequency fatigue') and recovery can take more than 1 day. Delayed force recovery is also seen in single muscle fibres from frog and mouse after fatigue induced by repeated, brief contractions. Evidence from our own and other laboratories indicates that the impairment is unlikely to result from metabolic changes and points to a defect in excitation-contraction coupling. We demonstrate that the likely site of failure is in the coupling between t-tubule depolarization and release of Ca2+ from the SR. The causative agent appears to be a localized increase in cytoplasmic Ca2+ which initiates some disruptive process, which can, however, be fully reversed, albeit slowly. Our experimental evidence does not support the involvement of Ca(2+)-activated proteases. Attempts to clarify the possible role of Ca(2+) activated lipases (phospholipase A2) and Ca2+/calmodulin have been hampered by side-effects of available inhibitors. Efforts to clarify how Ca2+ exerts its effects are continuing. PMID- 8729680 TI - Modulation of muscle contractility during fatigue and recovery by ATP sensitive potassium channel. AB - The activity of ATP-sensitive potassium channels of skeletal muscle is controlled by changes in the bioenergetic state of the cell. These channels are inactive in unfatigued muscle and become activated during fatigue. It has been postulated that ATP-sensitive potassium channels shorten the action potential duration, increase the potassium efflux and contribute to the decrease in force during fatigue. Although blocking ATP-sensitive potassium channels during fatigue prolongs the action potential duration and decreases the potassium efflux as expected, it does not affect the rate of fatigue development as observed from the decrease in tetanic force. Even though such results are not consistent with the hypothesis that ATP-sensitive potassium channels contribute to the decrease in force during fatigue, a reduced capacity of skeletal muscles to recover their tetanic force following fatigue is also observed when ATP-sensitive potassium channels are blocked during fatigue, suggesting that these channels have a myoprotective effect. It is thus possible that removing this myoprotection during fatigue results in deleterious effects which counteract the expected slower decrease in force. However ATP-sensitive potassium channel openers also fall to affect the rate of fatigue development. Therefore, the results obtained so far do not support the hypothesis that ATP-sensitive potassium channels contribute to the decrease in force during fatigue. PMID- 8729681 TI - Fuzzy space and control of Na+, K(+)-pump rate in heart and skeletal muscle. AB - Since intracellular Na+ activity (aiNa) is one important determinant of Na+ K(+) pump rate as well as excitability and the finely tuned contractility, it is surprising that the relation between aNa and pump rate reported by different authors has kQ5 varying between 10 and 40 mmol L-1. Other data also point to a variable relation between pump rate and aiNa. During stimulation of isolated rat soleus muscles at 2 Hz, ouabain-sensitive 86Rb uptake was increased in spite of the intracellular Na+ remaining unaltered. In isolated cardiomyocytes, a transient Na+. K(+)-pump current was observed upon activation by extracellular K+ in spite of good control of aiNa. Na(+)-loaded, isolated, sheep cardiac Purkinje fibres initially hyperpolarized over a period of up to 1 min upon activation of the Na+, K+ pump with no detectable change of aiNa. These examples are compatible with the existence of a micro-environment close to the membrane where diffusion is slower than in the rest of the cytosol, so that local aiNa may fluctuate or gradients may develop as visualized by Wendt-Gallitelli at al. (1993). We conclude that the reported relationships between Na+, K+, pump rate and aiNa in intact cells probably underestimate the true affinity of the Na+, K+ pump for Na+ due to a functional diffusion barrier beneath the sarcolemma, and also because of incomplete cell dialysis in whole-cell voltage clamp experiments. The Na+, K+ pump seems to be preferentially supplied with Na+ from the outside through neighbouring channels and transporters. PMID- 8729682 TI - The Na+, K+ pump in skeletal muscle: quantification, regulation and functional significance. AB - In skeletal muscle, the Na+, K+ pump is predominantly situated in the sarcolemma (1000-3500 pumps per microns 2). The total concentration can be determined in fresh or frozen biopsies (1-5 mg) using a 3H-ouabain binding assay. The values obtained have been confirmed by measurements of maximum ouabain suppressible Na+, K(+)-transport capacity in intact muscles as well as Na+, K(+)-ATPase-related enzyme activity in muscle homogenates. In the mature organism, the concentration of Na+, K+ pumps varies with muscle type and species in the range 150-600 pmol (g wet wt)-1 in rat and human muscle, the concentration increases markedly with thyroid status. Semi-starvation and untreated diabetes reduce the concentration by 20-48%. K+ deficiency leads to a downregulation of up to 75%. Both in animals and in humans, training increases the concentration of Na+, K+ pumps in muscle and inactivity leads to a downregulation. High-frequency stimulation elicits up to a 20-fold increase in the net efflux of Na+ within 10 s This is the major activation mechanism for the Na+, K+ pump, utilizing its entire capacity and possibly represents a drive on de novo synthesis of Na+, K+ pumps. A variety of hormones (insulin, insulin-like growth factor I, adrenaline, noradrenaline, calcitonin gene-related peptide, calcitonin, amylin) increase the rate of active Na+, K+ transport by 60-120% within a few minutes. This leads to a decrease in intracellular Na+ and hyperpolarization. In isolated muscles, where contractility is inhibited by high extracellular K(+)- such agents produce rapid force recovery. which is entirely suppressed by ouabain and closely correlated to the stimulation of K+ uptake and the decline in intracellular Na+. The observations support the conclusion that the Na+, K+ pump plays a central role in the acute recovery and maintenance of excitability during contractile activity. PMID- 8729683 TI - Na+, K(+)-pump activity and skeletal muscle contractile deficits in the spontaneously hypertensive rat. AB - Skeletal muscles in an animal model of genetic hypertension (the spontaneously hypertensive rat. SHR) exhibit significant deficits in contractile performance. These deficits appear to be unrelated to the rise in blood pressure. Slow-twitch soleus muscles show a decrease in specific muscle tension and a reduced resistance to muscle fatigue during prolonged contractile activity. We tested the hypothesis that the reduced fatigue resistance occurs as a consequence of an impaired ability to maintain or restore Na+ and K+ balance across the sarcolemma during repeated contractions. This may result from a genetically based increase in the Na+ permeability of SHR muscles, coupled with a reduction Na+, K+ pump capacity as the animals mature. Soleus muscles in adult SHR exhibit a significant increase in intracellular Na+ content and a significant decrease in intracellular K+ content at rest. B6RB+ uptake in Na(+)-loaded hypertensive muscles is 45% less than predicted from the number of ouabain-binding sites available. Activation of Na+, K+ pumps using adrenaline or insulin produces a significantly smaller hyperpolarization in hypertensive soleus than in control muscles. Control soleus muscles are hyperpolarized for at least 10 min after a 4 min period of high frequency activity, but hypertensive soleus muscles remain at resting polarity. Nonetheless, the number of ouabain-binding sites in hypertensive muscle is significantly greater than in control soleus, and binding affinities are similar. This apparent deficit in pump capacity might lead to a greater and more prolonged increase in extracellular K+ during repetitive contractions,and an associated decline in tension. Recently, we have been able to prevent the abnormal decrease in hypertensive soleus fatigue resistance by long-term treatment (8 weeks) with the Ca2+ blocker amlodipine. The therapy prevented or reversed the contractile deficits, but did not restore the responsiveness of the Na+, K+ pump to hormonal stimulation. The current data suggest that both a reduction in Na+, K(+)-pump capacity and changes in Ca2+ distribution play a role in the development of contractile deficits in hypertensive muscles. PMID- 8729684 TI - Ion gradients and contractility in skeletal muscle: the role of active Na+, K+ transport. AB - Intensive contractile activity is associated with a significant net loss of K+ and a comparable gain of Na+ in the working muscle fibres. This leads to an increase in the interstitial and T-tubular K+ concentration and to a decrease in the T-tubular Na+ concentration. It is well established that the exposure of muscles to high extracellular K+ or low extracellular Na+ inhibits contractile performance. More importantly, the combination of high extracellular K+ and low extracellular Na+ has a much more pronounced inhibitory effect on force than the sum of the individual effects of the two ions. The inhibitory effects of high extracellular K+ or low extracellular Na+ can be alleviated within 5-10 min by acute hormonal stimulation of the Na+, K+ pump. In contrast, reductions in the capacity for active Na+, K+ transport by pre-incubation of isolated muscles with ouabain or by prior K+ depletion of the animals significantly decreases contractile endurance during high-frequency electrical stimulation. Thus, muscles from K(+)-depleted rats exhibiting a 54% reduction in Na+, K+ pump concentration showed a 110% increase in force decline during 30 s of 60 Hz stimulation. Reducing the Na+, K+ pump capacity to a similar extent by pre-incubation with ouabain led to a comparable decrease in endurance. Moreover, reductions in the Na+, K+ pump capacity were associated with an increased intracellular accumulation of Na+ during electrical stimulation. These observations support the notion that excitation-induced decreases in Na+, K+ gradients contribute to fatigue during intensive exercise and suggest that the capacity for active Na+, K+ transport is a determining factor for contractile endurance. PMID- 8729685 TI - Potassium and fatigue: the pros and cons. AB - A general finding is that muscle activity leads to potassium fluxes across the muscle membrane as well as to muscle fatigue, defined as a reduction in the force generating capacity of the muscle. However, much controversy exists regarding the causal role of potassium in fatigue development. The experimental model used is decisive in this context, e.g. whether we study intact in vivo organs in situ with voluntary contractions and reflex feedback for cardiorespiratory regulation, or whether we study in vitro isolated muscles or cells-or even skinned fibres. In the latter models, clear evidence has been presented that Ca2+ is the variable significant for force development and that K+ may be ignored. However, in the in situ situation the limiting link in the chain leading to muscle contraction may be one preventing the Ca2+ release from taking place. The sites are the triads, T tubules, and the surface membrane. The function of the latter two regarding action potential amplitude and propagation depends on [K+] gradients. Again, conflicting results exist regarding the electrophysiological changes and [K+] in fatigue. The activity pattern must then be taken into consideration. During high intensity (high-frequency) activity the increased interstitial [K+] has been demonstrated to relate to fatigue, while in low-intensity fatiguing protocols, the T-tubule may be the limiting site. This fits with the concept of interstitial [K+] playing an essential role as a regulatory feedback mechanism, e.g. adjusting muscle blood flow to the metabolic load during muscle activity. PMID- 8729686 TI - High-and low-frequency fatigue revisited. AB - Changes in excitation-contraction coupling have long been recognized as possible causes of the failure in function which occurs in fatigued muscle. High-frequency fatigue is characterized by an excessive loss of force at high frequencies of stimulation and rapid recovery when the frequency is reduced. Frequencies in excess of 50 Hz are rarely seen with voluntary activation of human muscle, and for this reason there has been some doubt as to whether high-frequency fatigue is a significant feature of normal activity. Recent experiments have shown that with 30 Hz stimulation there is a more rapid loss of force if the muscle is held isometric in a shortened position and the fatigue is rapidly reversed if the muscle is re-extended, even under ischaemic conditions. These findings are consistent with the accumulation of K+ in the t-tubules and interfibre spaces of the muscle. Low-frequency fatigue is characterized by a relative loss of force at low frequencies of stimulation and a slow recovery over the course of hours or even days. There is evidence from intracellular measurements that low-frequency fatigue is due to a reduction in Ca2+ release. However, there is a possibility that in the fatiguing exercise, the end sarcomeras of the fibre overextend and damage those in the middle section of the fibre. In this situation the active sarcomas would be working at a shorter length than predicted from the overall fibre length and the force-frequency curve will be shifted to the right. Measurements of the length-tension relationship of muscles damaged by stretching are consistent with this happening. PMID- 8729687 TI - Muscular function, metabolism and electrolyte shifts during prolonged repetitive exercise in humans. AB - Marked functional changes occur in human skeletal muscle during prolonged repetitive exercise. The maximum voluntary contraction force (MVC) falls gradually and may reach 50% of control within 30-60 min. The twitch tension declines faster and to a larger extent. During repetitive submaximal isometric contractions the rate of relaxation increase progressively, in parallel with an increased energy cost of contraction. These functional changes are all slowly reversed in the post-exercise period, as indicated by only minor changes over the first 30 min of recovery. Minor changes in substrates and metabolites, together with the slow rate of recovery, indicate that the alterations in contractile properties and energetics are independent of these metabolic factors. Alternative explanations may be related to electrolyte shifts over the sarcolemma or between cellular compartments. The total loss of K+ is small, and could not be detected by analysis of muscle biopsies. Only a slight initial rise in muscle content of calcium was found. The available data indicate that the increased energy cost of contraction is not connected to mitochondrial dysfunction, which might be caused by calcium accumulation. Rather, it seems that the ratio of ATP utilization to force is increased and this could possibly be connected to this faster relaxation rate. Considering the low excitation rates during submaximal voluntary contractions, each motor unit generates an oscillating force closely associated with Ca2+ fluctuations between SR and cytosol. Increased relaxation rats might be caused by faster reuptake of Ca2+ into the SR, and this could contribute to the faster ATP turnover. PMID- 8729688 TI - K+ balance in humans during exercise. AB - Onset of exercise leads to a sudden increase in [K+] in venous plasma from the exercising muscles. Delayed by about 10 s. the arterial [K+] rises nearly at the same rate as the venous concentration. At exercise intensities below 100% of Vo2max, both venous and arterial [K+] stabilize at a steady-state value. At higher intensities, venous and arterial plasma [K+] continue to rise until exhaustion. During the first 5 min of exercise the contracting muscles always lose K+, with a peak in loss rate after 1-2 min. During steady state, the loss rate is minimized or may even be reduced to zero. The loss is caused by an exercise-induced afflux of K+ from the contracting cells which exceeds the exercise-induced influx mediated by the Na, K pump. The Na, K pump is stimulated by catecholamines in vitro and in resting tissue in vivo. However, the loss rate of K+ from steady-state exercising muscles does not show any increase during beta -adrenergic blockade or decrease during beta-adrenergic stimulation. This is probably due to a compensatory change in intracellular [Na+]. During low exercise intensity, arterial [K+] does not increase after 1-2 min. while the exercising muscles lose K+, showing that the extracellular pool of K+ is redistributed. During beta-adrenergic blockade this redistribution is impaired so that the rise in plasma [K+] is accentuated. Conversely alpha-adrenergic blockade reduces the exercise-induced hyperkalaemia. Hence, the adrenergic system plays an important role in regulation of whole-body K+ balance during exercise but its significance in exercising muscles is not clear. PMID- 8729689 TI - Role of potassium in the regulation of systemic physiological function during exercise. AB - In exercise, potassium (K+) is released from contracting muscle predominately through K+ channels associated with the repolarization phase of the action potential. Increases in extracellular K+ are directly related to increases in metabolic rate and may reach concentrations as high as 8-9 mM in the arterial blood during exhaustive work. Exercise-induced hyperkalaemia has been implicated in several physiological processes, in particular skeletal muscle fatigue, hyperaemia, pressor reflex, arterial chemosensitivity and myocardial stability. There is no direct evidence to show that hyperkalaemia causes muscle fatigue, although raised extracellular [K+] may contribute to fatigue during prolonged tetani by depressing the propagation of the action potential down the t-tubule system, thus impairing the release of Ca2+ from the sarcoplasmic reticulum. The vasodilating properties of K+ may transiently contribute to the early phase of exercise hyperaemia and interact synergistically with other vasoactive substances to cause relaxation by hyperpolarizing K+ channels in vascular smooth muscle. Hyperkalaemia has been implicated in the regulation of arterial blood pressure through activation of the muscle afferent reflex where potassium-depolarized C fibres may contribute to a reflex increase in arterial blood pressure. K+ can also increase ventilation and the sensitivity of the ventilatory response to hypoxia through direct excitation of the arterial chemoreceptors. Finally, to maintain myocardial electrical stability in exercise, there is a beneficial interaction between raised K+ and catecholamines on the heart, so that when they combine, each offsets the other's deleterious effects. PMID- 8729690 TI - Role of skeletal muscle sodium pumps in the adaptation to potassium deprivation. AB - Skeletal muscle is specialized to lose K+ to the extracellular fluid during potassium deprivation which buffers the fall in plasma K+ concentration. While it remains to be determined whether K+ efflux from muscle is altered during K+ deprivation, active K+ uptake driven by sodium pumps is significantly depressed. The activity of sodium pumps in skeletal muscle does not increase during K+ depletion despite elevated intracellular Na+, a strong stimulus to increase activity in other cells. There is a decrease in the total pool size of sodium pump alpha beta heterodimers during potassium deprivation. The alpha 2 (not the alpha 1) sodium pump isoform is specifically decreased and beta 1 and/or beta 2 decreases in a muscle-fibre-dependent manner. The specific loss of K+ from skeletal muscle is probably a consequence of the fact that the alpha 2 isoform predominates in this tissue. In tissues such as heart, where alpha 2-type pumps are only a minor fraction of the sodium pumps, the activity of the ubiquitous alpha 1 isoform maintains intracellular Na+ and K+ at control levels, despite the fact that alpha 2 levels decrease by 50%. Analysis of the time course of change in alpha 2 mRNA vs. protein during K+ deprivation indicates that there is both a decrease in alpha 2 synthesis and an increase in alpha 2 degradation. The apparent time-lag during potassium deprivation between the early decreases in both surface alpha 2-type sodium pump number (assessed by 3H-ouabain binding) and intracellular K+, and the later decrease in total pool size of alpha 2, suggests the hypothesis that there may be an early internalization of alpha 2 sodium pumps to endosomal pools, followed by a degradation of these internalized pumps, contributing to the decrease in total alpha 2 pool size. The signals mediating this specific response to hypokalemia, and those mediating the restoration of muscle K+ stores remain to be determined. PMID- 8729691 TI - Effects of K+, Mg2+ deficiency and adrenal steroids on Na+, K(+)-pump concentration in skeletal muscle. AB - Animal studies have shown that deficiency of K+ is associated with a reduction in the concentration of Na+, K+ pumps in skeletal muscle, and that this reduction is closely correlated with the reduction in the muscle K+ concentration. Furthermore, animals deficient in Mg+ show a downregulation of the Na+, K(+)-pump concentration, but this seems to be secondary to the concomitant K+ deficiency, which often accompanies Mg2+ deficiency. Measurements on skeletal muscle biopsies from patients who had been in long-term treatment with diuretics showed that 55% had reduced concentrations of both K+ and Mg2+, and that this was associated with a reduction in the concentration of Na+, K+ pumps. Furthermore, the Na+, K(+) pump concentration correlated significantly with both muscle K+ and Mg2+, suggesting that the downregulation of the Na+, K+ pumps was related to the loss of K+, as predicted from the animal experiments. In accordance with this, normalization of muscle K+ and Mg2+ in response to oral Mg2+ supplementation, resulted in a restoration of the Na+, K+ pumps. Apart from thyroid hormone, which is another major regulator for the Na+, K(+)-pump concentration, other hormones may be of importance. It is well known that adrenal steroids control the synthesis of Na+, K+ pumps in the kidney and heart. Recently, treatment with dexamethasone was found to increase the Na+, K(+)-pump concentration in rat skeletal muscle. The increase was found in EDL, soleus, gastrocnemius and diaphragm muscles, and amounted to 23-52%. In contrast, treatment with aldosterone induced a decrease in the Na+, K(+)-pump concentration, which was closely correlated to the reduced K+ content of the muscles. The results indicate that in skeletal muscle, high doses of glucocorticoids upregulate the concentration of Na+, K+ pumps, whereas mineralocorticoids induce a downregulation which is secondary to the concomitant K+ deficiency. PMID- 8729692 TI - Developmental regulation of cation pumps in skeletal and cardiac muscle. AB - The prenatal and early postnatal periods are critical stages during which long term development can be affected. For example, retardation of growth during these periods is closely linked to the occurrence of adult degenerative diseases. Appropriate development of muscle is essential for numerous functions, including movement, posture, thermogenesis, breathing and maintenance of the circulation. Defects in normal muscle development could thus impair any of these functions in the neonate and may also have long-term consequences for the health of the individual. Central to normal muscle structure and function is the appropriate development not only of the sarcomeric proteins but also of the sarcolemma, transverse-tubules, sarcoplasmic reticulum and associated membrane-bound ATPases. Long-term regulation of these ATPases is by changes in their concentration, whereas short-term regulation is mediated by alterations in enzyme activity. This review focuses on changes in total concentrations of Na+, K+, and Ca(2+)-ATPases during prenatal and postnatal life, in functionally diverse muscles of mammalian species born at different stages of maturity. Both these cation pumps belong to multigene families and changes in relative abundance of their specific isoforms are also considered because they may have important consequences for contractile performance during distinct stages of development. Finally, potential regulatory mechanisms which alter markedly during normal ontogeny are discussed. These include intrinsic factors such as hormones and contractile activity, extrinsic factors such as nutrition and environmental temperature, and interactions between these variables which are known to be especially important during postnatal development. PMID- 8729693 TI - Effects of thyroid hormones on contractility and cation transport in skeletal muscle. AB - Skeletal muscle is one of the major target organs for thyroid hormone. The muscles most commonly affected are those used during prolonged effort (slow twitch muscles). One of the major clinical features is the shortening of the Achilles-tendon reflex time in hyperthyroidism and its prolongation in hypothyroidism. Most of the peripheral effects of the thyroid hormones can be ascribed to the action of triiodothyronine (T2), which is produced by de iodination of thyroxine (T4) in liver and kidney. From the plasma, T3 is actively transported into skeletal muscle. The Ca2+ ATPase in skeletal muscle is responsible for removal of Ca2+ ions from the cytosol into the sarcoplasmic reticulum (SR) during relaxation, and the Na+, K+ ATPase in the plasma membrane is responsible for restoration of the membrane potential after excitation. The concentrations of Ca2+ ATPase and Na+, K+ ATPase in rat skeletal muscle have been shown to increase four- and 10-fold, respectively, in the transition from the hypothyroid to the hyperthyroid state. In humans, a linear correlation between the Na+, K+ ATPase concentration of skeletal muscle and the free T4 index was established. Significant effects of T3 on Ca2+ ATPase and Na+, K+ ATPase can be detected 24 h after a single injection. These effects are mediated by increased production of mRNA for the respective proteins, initiated by binding of T3 to nuclear receptors. Passive fluxes of Ca2+, Na+ and K+ also show a significant rise after T3 treatment. The increase in passive fluxes of Na+ and K+ can be detected before the rise in the concentration of Na+, K+ ATPase, suggesting that T3. In addition to its nuclear effects, may have a direct effect on the plasma membrane. Apart from their significance for muscle function in thyroid disease, the changes in Ca2+ ATPase and Na+, K+ ATPase will be important in other conditions where T3 and T4 levels show dramatic changes, i.e. during postnatal development, starvation and undernutrition, as well as in non-thyroidal illness (low-T3 syndrome). PMID- 8729694 TI - Effects of training on potassium, calcium and hydrogen ion regulation in skeletal muscle and blood during exercise. AB - Ionic regulation is critical to muscle excitation, contraction and metabolism, and thus for muscle function during exercise. This review focuses on the effects of training upon K+, Ca2+ and H+ ion regulation in muscle and K+ regulation in blood during exercise. Training enhances K+ regulation in muscle and blood and reduces muscular fatiguability. Endurance, sprint and strength training in humans induce an increased muscle Na+, K+ pump concentration, usually associated with a reduced rise in plasma [K+] during exercise. Although impaired muscle Ca2+ regulation plays a vital role in fatigue, little is known about possible training effects. In rat fast-twitch muscle, overload-induced hypertrophy and endurance training were associated with reduced sarcoplasmic reticulum Ca2+ uptake, consistent with fast-to-slow fibre transition. In human muscle, endurance and strength training had no effect on muscle Ca2+ ATPase concentration. Whilst muscle Ca2+ uptake, release and Ca2+ ATPase activity were depressed by fatigue, no differences were found between strength athletes and untrained individuals. Muscle H+ accumulation may contribute to fatigue during intense exercise and is also modified by sprint training. Sprint training may increase muscle Lac- and work output with exhaustive exercise, but the rise in muscle [H+] is unchanged or attenuated, indicating a reduced rise in muscle [H+] relative to work performed. Muscle buffering capacity can be dissociated from this improved H+ regulatory capacity after training. Thus, training enhances muscle and blood K+ and muscle H+ regulation during exercise, consistent with improved muscular performance and reduced fatiguability; however, little is known about training effects on muscle Ca2+ regulation during contraction. PMID- 8729695 TI - Stimulation of Na+, K(+)-pump activity in skeletal muscle by methylxanthines: evidence and proposed mechanisms. AB - Evidence is presented to support the hypothesis that submillimolar concentrations of methylxanthines stimulate Na+, K(+)-ATPase activity in skeletal muscle. Administration of methylxanthines to skeletal muscle results in plasma membrane hyperpolarization and increased rates of K+ uptake and Na+ efflux. These effects are both dose- and time-dependent and inhibited by blockers of the Na+, K+ ATPase. The mechanisms for stimulation of Na+, K(+)-ATPase activity and the signal transduction pathways are not known. The methylxanthine concentrations required for stimulation of Na+, K(+)-ATPase activity are less than those required to cause a 50% inhibition of phosphodiesterase activity, and therefore increases in cyclic AMP due to inhibition of the enzyme are not involved. Possible mechanisms by which methylxanthines may increase Na+, K(+)-ATPase activity include; (1) a role for increased intracellular [Ca2+]; (2) Ca2+ or adenosine-receptor-mediated increases in intracellular cyclic AMP; and (3) a direct action of methylxanthines on the Na+, K+ ATPase. PMID- 8729696 TI - Ion transport in human skeletal muscle cells: disturbances in myotonic dystrophy and Brody's disease. AB - After excitation of skeletal muscle, the disturbed ion homeostasis is restored by Na+, K+ ATPase of the sarcolemma and Ca2+ ATPase of the sarcoplasmic reticulum (SR). Contrary to Na+, K+ ATPase, the concentration and isoenzyme distribution of SR Ca2+ ATPase in human skeletal muscle depend on fibre type and age. In cultured human muscle cells the concentration and activity of Na+, K+ ATPase and SR Ca2+ ATPase increase with maturation. In skeletal muscle and cultured muscle cells of patients suffering from myotonic dystrophy (MyD), the activity and the concentration of both Na+, K+ ATPase and SR Ca2+ ATPase are decreased by about 40%. In addition, we measured in cultured MyD muscle cells at rest an increased cytosolic Ca2+ concentration ([Ca2+]i) caused by active voltage-operated Ca2+ channels, which are inactive in resting control cells. However, the restoration of a stimulus-induced Ca2+ transient is unaffected. A differentiation-related disturbance of membranes or a modulation defect of membrane proteins may play a role in MyD. In skeletal muscle and cultured muscle cells of patients suffering from Brody's disease, which is characterized by impaired muscle relaxation, the SR Ca2+ ATPase activity is reduced by about 50%, but the concentrations of total SR Ca2+ ATPase and the predominant SERCA1 isoform are normal. Diseased muscle cells show a delayed restoration of [Ca2+]i after stimulation, which might be explained by structural modifications of SERCA1. Reduction of the Ca2+ release by drugs balances the excitation-relaxation cycle of the pathological cells. PMID- 8729697 TI - Lactate/proton co-transport in skeletal muscle: regulation and importance for pH homeostasis. AB - In skeletal muscle, intracellular pH is more alkaline than would be predicted if H+ were passively distributed across the sarcolemma. Therefore, the passive influx of H+ must be counteracted by transport processes mediating H+ afflux. In resting skeletal muscle, these transport processes are Na+/H+ exchange and bicarbonate-dependent systems. During periods of high energy demand, skeletal muscle produces large amounts of lactic acid. The internal accumulation of lactic acid reduces pH, which may cause fatigue. It is therefore important for muscle cells to be able to regulate pH during and after activity. A part of the accumulated lactate and H+ is metabolized, but a considerable fraction is released from the cell. The efflux of H+ and lactate might be mediated by the lactate/proton co-transport system found in almost all cell types in the body. The role of lactate/proton co-transport in pH regulation has been studied both with intact cells and with sarcolemmal vesicles. In intact cells, inhibitors of lactate/proton transport have been shown to accelerate the development of fatigue, and to delay the recovery after activity. A comparison with vesicles has demonstrated that, at low pH, and with a high lactate concentration, the capacity for H+ removal is higher via the lactate/proton co-transport system than via the sum of the Na+/H+ exchange and bicarbonate-dependent exchange systems. Therefore, the carrier-mediated lactate/proton efflux is of major importance for pH regulation in connection with muscle activity. The lactate/proton transport system has been shown to undergo long-term changes depending on the level of physical activity. The capacity of the system was enhanced after intense training or chronic stimulation, and reduced after denervation. It is concluded that the lactate/proton transport system is of major importance for pH regulation in skeletal muscle, and that changes in the amount of transporters are one of the many adaptations to physical activity. PMID- 8729698 TI - Ion channels in the sarcoplasmic reticulum of striated muscle. AB - This review provides a summary of current concepts about the structure and single channel properties of ryanodine receptor calcium release channels and counter ion channels that facilitate Ca2+ release and reuptake by the sarcoplasmic reticulum. Some recent results, obtained with single ryanodine receptor ion channels incorporated into lipid bilayers from terminal cisternae vesicles of rabbit skeletal muscle and sheep ventricular myocardium, are described. The ryanodine receptor is the major Ca2+ release channel in skeletal and cardiac muscle and has been studied in far greater detail than other sarcoplasmic reticulum ion channel proteins. Several ryanodine receptor genes have been cloned and sequenced, and isoforms of the protein have been detected in muscle and in endoplasmic reticulum of brain and many other tissues from mammals, lower vertebrates, nematodes and drosophila. The proteins from all species are tetramers of a peptide with a molecular mass of approximately equal to 560 kDa, containing approximately equal to 5000 amino acids, with a similar maximum single-channel conductance of 500-800 row S for monovalent cations at 250mM. Results presented here include: Ca2+ activation and adaptation of activity in skeletal ryanodine receptors with rapid changes in [Ca2+] controlled by perfusion; activation by FK506 and regulation of cooperative gating of skeletal ryanodine receptor channel activity by FK506 binding proteins; activation and block of cardiac ryanodine receptors by addition of reactive disulphides and by bilayer voltage. Effects of phosphorylation, calmodulin, triadin, calsequestrin and interactions with the alpha 1 subunit of the dihydropyridine receptor on ryanodine receptor activity are summarized. Potassium and chloride channels in skeletal muscle sarcoplasmic reticulum, are described. PMID- 8729699 TI - Calcium uptake and release modulated by counter-ion conductances in the sarcoplasmic reticulum of skeletal muscle. AB - The sarcoplasmic reticulum (SR) plays the central role in regulating the free myoplasmic Ca2+ level for the contractile activation of skeletal muscle. The initial stages of the voltage-controlled Ca2+ release mechanism are known in molecular detail. However, there is still very little known about the later stages of Ca2+ uptake and total Ca2+ turnover in the contraction-relaxation cycle under normal physiological conditions or under conditions influenced by fatigue or disease. Ca2+ uptake and release are both accompanied by "counter-ion' movements across the SR membrane which prevent or reduce the generation of SR membrane potentials and balance for electroneutrality in the SR lumen. The SR membrane is permeable for the cations K+, Na+, H+ and Mg2+ and the anion Cl-. Using electron-probe X-ray microanalysis. It has been shown that during tetanic stimulation the Ca2+ release was mainly balanced by uptake of K+ and Mg2+ leaving a charge deficit that was assumed to be neutralized via H+ ion or organic counter ion movement. The low time resolution of electron-probe X-ray microanalysis leaves the possibility of other transient concentration changes in the SR, e.g. for Cl- ions. Possible physiological roles of the SR counter-ion conductances can be tested using skinned muscle fibre preparations with intact sarcoplasmic reticulum and removed or chemically permeabilized outer sarcolemma. In skinned fibres, the SR K+ conductance can be effectively reduced with SR K+ channel blockers such as 4-aminopyridine, tetraethylammonium and decamethonium. Interestingly, these blockers increase Ca2+ loading as well as Ca2+ release, whereas other less specific blockers, such as 1.10-bis-quanidino-n-decane, seem to reduce Ca2+ release, possibly also via blocking Ca2+ release channels. Thus, it seems very important also to test the effects of counter-currents carried by K+, Mg2+, H+ or Cl- ions on intact and voltage-clamped single-fibre preparations. PMID- 8729700 TI - Membrane abnormalities and Ca homeostasis in muscles of the mdx mouse, an animal model of the Duchenne muscular dystrophy: a review. AB - Muscles of the mdx mouse lack dystrophin, a cytoskeletal protein. Mdx fibres exhibit an increased fragility to hypo-osmotic shock and to forced lengthening, an abnormal opening time of stretch-sensitive calcium channels. The question of a chronic elevated [Ca2+]i value is a matter of controversy. We have analysed Ca homeostasis in smooth and skeletal muscles from the adult mdx mouse. The wall of the vas deferens was loaded with the fluorescent Ca indicator Fura-2-AM (cell diffusible). Resting [Ca2+]i was measured after changes of the electrochemical potential for Ca2+ and after KCl or electrical stimulations. In no instance was a difference observed between these and similar muscles from control mice. Single striated fibres were isolated by collagenase treatment of the flexor digitorum brevis muscle and loaded with Fura-2-AM. The value of resting [Ca2+]i was measured using an in situ calibration procedure which took account of Ca buffering by Fura-2. A chronic increase of cytosolic Ca2+ was not confirmed. The expression of the intracellular Ca-binding protein, parvalbumin, was measured. It increased by about threefold in fast mdx muscles (tibialis anterior) but remained undetectable in the soleus. It is hypothesized that parvalbumin helps to maintain [Ca2+]i within normal values. This hypothesis will be discussed in connection with dystrophy phenotypes in mutant dogs and in human patients. PMID- 8729701 TI - Mechanisms underlying changes of tetanic [Ca2+]i and force in skeletal muscle. AB - Force development in skeletal muscle is driven by an increase in myoplasmic free [Ca2+]i ([Ca2+]i) due to Ca2+ release from the sarcoplasmic reticulum (SR). The magnitude of [Ca2+]i elevation during stimulation depends on: (a) the rate of Ca2+ release from the SR; (b) the rate of Ca2+ uptake by the SR; and (c) the myoplasmic Ca2+ buffering. We have used fluorescent Ca2+ indicators to measure [Ca2+]i in intact, single fibres from mouse and Xenopus muscles under conditions where one or more of the above factors are changed. The following interventions resulted in increased tetanic [Ca2+]i: beta-adrenergic stimulation, which potentiates the SR Ca2+ release; application of 2.5-di(tert-butyl)-1,4 benzohydroquinone, which inhibits SR Ca2+ pumps; application of caffeine, which facilitates SR Ca2+ release and inhibits SR Ca2+ uptake; early fatigue, where the rate of SR Ca2+ uptake is reduced; acidosis, which reduces both the myoplasmic Ca2+ buffering and the rate of SR Ca2+ uptake. Reduced tetanic [Ca2+]i was observed in late fatigue, due to reduced SR Ca2+ release, and in alkalosis, due to increased myoplasmic Ca2+ buffering. Force is monotonically related to [Ca2+]i but depends also on the myofibrillar Ca2+ sensitivity and the maximum force cross bridges can produce. This is clearly illustrated by changes of intracellular pH where, despite a lower tetanic [Ca2+]i, tetanic force is higher in alkalosis than acidosis due to increases of myofibrillar Ca2+ sensitivity and maximum cross bridge force. PMID- 8729702 TI - Social physique anxiety, body esteem, and social anxiety in bodybuilders and self reported anabolic steroid users. AB - Social physique anxiety, upper body esteem, social anxiety, and body dissatisfaction were examined among self-reported anabolic steroid (AS)-using bodybuilders, non-using bodybuilders, athletically active exercisers, and non exercising individuals. Results indicate that: (a) the AS-using bodybuilder group had significantly lower levels of social physique anxiety than non-user groups, (b) AS-using bodybuilders have significantly higher upper body strength ratings than do non-users (non-using bodybuilders, athletically active exercisers, and non-exercisers), and (c) no differences among groups on a measure of social anxiety. Implications for further research and intervention are discussed in terms of the differences between anabolic steroids and other substances of abuse. PMID- 8729704 TI - Predictors of neuropsychological impairment in alcoholics: antisocial versus nonantisocial subtypes. AB - The role of demographic, perinatal/developmental, and acquired subject characteristics in determining neuropsychological (NP) performance was investigated in 22 alcoholics with Antisocial Personality Disorder (ASPD) and 84 non-ASPD alcoholics. Results of stepwise multiple regression analyses revealed that in ASPD subjects, poor NP performance was predicted by less education, childhood symptoms of Conduct Disorder, drinks per drinking day, and history of head injury, accounting for 80% of the explained variance (p < .0001). In non ASPD subjects, NP performance was predicted by self-reported history of diagnosed Attention Deficit Disorder, Verbal Learning Disability, and symptoms of Nonverbal Learning Disability, accounting for 24% of the explained variance (p < .0001). These results suggest the presence of potentially different lifelong paths to NP impairment among ASPD and non-ASPD alcoholics. Further exploration of the multivariate predictors of neuropsychological performance in subgroups of alcoholics is warranted. PMID- 8729703 TI - Investigation of mechanisms linking depressed mood to nicotine dependence. AB - The present study examined the cognitive-behavioral linkages between depressed mood and level of nicotine dependence in smokers seeking smoking cessation treatment. Prior to treatment, 202 subjects completed validated self-report measures of smoking history, depressive symptomatology, "self medication" processes, "learned helplessness" processes, and nicotine dependence. Results revealed that 48% of the study population scored in the "depressed" range on the Center for Epidemiologic Studies (CESD) depression scale. Further, these smokers reported significantly higher levels of nicotine dependence than other nondepressed smokers. Depressed and nondepressed smokers did not differ with respect to several cognitions related to learned helplessness theory. However, depressed smokers were more likely to report "self medication" processes (i.e., negative affect reduction smoking and stimulation smoking). In addition, multivariable regression and path analyses suggested that negative affect reduction smoking and stimulation smoking are sequential mediators of the depression-nicotine dependence relationship. These results underscore the need to screen for depressive symptomatology among smokers seeking treatment, and to develop cessation treatments that are tailored to the needs of depressed smokers. PMID- 8729705 TI - Smoking cessation in pregnancy: the effect of hypnosis in a randomized study. AB - At Buskerud Central Hospital in Norway, a county hospital with 2000 deliveries per year, an intervention study, using hypnosis, aimed at smoking cessation and reduction among pregnant women still smoking around 18th week of pregnancy was carried out during the period 1992-1993. Two sessions (each lasting 45 minutes) using conventional induction into a superficial nonsomnabulistic stage of trance were performed. A tape was played, encouraging the pregnant woman's wish to quit smoking and her capacity to do so. Relaxation techniques together with self hypnotic methods were introduced to combat craving. Changes in smoking pattern were investigated at delivery time. No significant effect on smoking cessation or smoking reduction was obtained (p > 0.05). We recorded a 10% quit rate in both intervention group and control group, and 42% and 31% reduced their smoking in the intervention group and control group, respectively. PMID- 8729706 TI - An investigation of potential time effects in retrospective reports of cocaine relapses. AB - Retrospective reports are widely used in social science research, such as studies of the relapse process. There is evidence, however, that such reports can be affected by a number of factors, including the passage of time. The goal of this study was to determine whether retrospective reports of cocaine relapses were influenced by the amount of time that had elapsed between the onset of the relapse and when the relapse interview was administered. A structured cocaine relapse interview was used to gather information on experiences prior to relapse and attributions for relapse. Evidence of linear and/or nonlinear time effects was found in two of the six subscales assessing experiences prior to relapse (Social Pressure to Use Drugs and Sensation Seeking), and there was evidence of logarithmic time effects in four of the six subscales that assessed attributions for relapses. These findings indicate that the amount of time between the onset of a relapse and when the subject is interviewed can influence reports of experiences prior to relapse and attributions for relapse. However, the magnitude of the time effects was generally small. PMID- 8729707 TI - Adolescents' promotion of nonsmoking and smoking. AB - Previous studies of the influence of friends on adolescent smoking have adopted the approach of asking adolescents about their friends. However, studies in which the friends were also approached have shown that adolescents' reports of friends behavior were not accurate (more closely resembled their own behavior). In order to gauge the level of active social influence among adolescents we asked 14-15 year old students what they did to influence others not to smoke or to smoke. Across two surveys, a quarter of the Form 4 students were promoting antismoking messages or activities and 3% were promoting smoking by encouraging or forcing friends, siblings and acquaintances to smoke. Relatively more positive beliefs about smoking identified those students who actively promoted smoking whereas some positive beliefs about the adverse effects of cigarette advertising tended to identify those who were actively antismoking. Use of this approach to examine the issue of social influence provided information about the antismoking activities which students conducted, and could be used to further examine which messages students do not believe or do not use. For example, students believed that smoking reduces fitness but did not promote this message to others, and the reasons for such disparity need to be investigated and should be addressed in the content of prevention programs. PMID- 8729708 TI - The clinical significance of binge eating in an obese population: support for bed and questions regarding its criteria. AB - The present study examined the prevalence of binge eating disorder (BED) among 192 obese individuals seeking treatment in a university-based eating disorders program. Subjects responded to a self-report questionnaire on the presence and severity of binge eating and were divided into two groups: (a) 48 non-binge eaters and (b) 144 binge eaters, 72 of whom met criteria for BED. Binge eaters reported earlier onset of obesity, higher levels of psychopathology, and less competence in relation to weight control. Comparison between the 72 individuals diagnosed with BED and the 72 binge eaters not meeting BED criteria indicated that the former group evidenced more severe binge eating and a higher level of depression. Given the differences across a range of psychological variables between binge eaters and non-binge eaters, these findings support the clinical and research value of BED as a new diagnostic category. However, they also indicate that severity of binge eating, regardless of frequency and associated behavioral features, is the most important distinguishing characteristic. Further research is needed to determine the most appropriate threshold for BED diagnosis. PMID- 8729709 TI - Latent transition analysis to the stages of change for smoking cessation. AB - A key construct of the Transtheoretical Model of Behavior Change is the stage of change (Precontemplation, Contemplation, Preparation, Action, and Maintenance) (Prochaska & DiClemente, 1983, 1986). This study applies Latent Transition Analysis (LTA; Collins & Wugalter, 1992; Collins, Wugalter, & Rousculp, 1991) to movement through the stages of change for smoking cessation. LTA is an alternative method of assessing categorical outcome that is sensitive to change, diagnostic, and allows for the comparison of alternative models of change. The sample consisted of 545 reactively recruited smokers and former smokers who were assessed five times over a two year period. LTA was used to compare three specified models. The model that fit the data best indicates that in a six month time frame both progression and regression among the stages takes place as well as two stage progression. Examination of the probability of movement among the stages revealed three findings consistent with the Transtheoretical Model. First, movement through the stages is not always linear. Second, the probability of forward movement was greater than backward movement. Third, the probability of moving to adjacent stages was greater than the probability of two stage progression. PMID- 8729710 TI - Barriers to help-seeking for change in drinking: a gender-focused review of the literature. AB - Women with drinking problems are less likely than men to utilize alcohol-specific treatment services; they are comparatively more likely to seek help in general health and mental health care settings. These patterns in service utilization may be attributable, in part, to gender differences in barriers or disincentives to seeking and receiving care for alcohol abuse. In this paper we provide a gender focused review of research on barriers to help-seeking for addictions behavior change. Two major categories of barrier, involving client and treatment program characteristics, appear to have a greater impact on service utilization of women than men. Insights gained from barriers literature provide valuable direction for future research and suggest applications in clinical service provision and treatment planning. PMID- 8729711 TI - Smoking withdrawal symptoms in response to a trauma-related stressor among Vietnam combat veterans with posttraumatic stress disorder. AB - This study investigated the relationship between a trauma-related stressor and smoking withdrawal symptoms in 25 male Vietnam combat veterans with posttraumatic stress disorder (PTSD) using a within-subjects design. All subjects were smokers. The stressor involved a modified Stroop task, in which the veterans color-named either anxiety-related or neutral control words. Anxiety-related words produced more withdrawal symptoms than neutral control words, including increased craving, negative affect symptoms, somatic symptoms, and lack of alertness. PMID- 8729712 TI - Weight control self-efficacy types and transitions affect weight-loss outcomes in obese women. AB - Matching obesity treatments to heterogeneous clients is a recent evolution in the development of more effective weight-control programs, yet most interventions emphasize the external features of treatments rather than the internal belief structures of individuals. The purpose of this study was to determine whether Q methodology would identify distinct types of weight-control self-efficacy beliefs in obese women that would be linked to outcomes of a weight-loss program. Fifty four women (45 +/- 9 yrs, Mean +/- SD) 136 +/- 10% over ideal body weight participated in a 9-month nutritional/behavioral weight loss program. Two major self-efficacy categories emerged through factor analysis of Q sorts: assureds and disbelievers. The assureds (n = 28) had the strongest self-efficacy beliefs and at baseline reported significantly (p < .01) greater self-esteem and less depression than the disbelievers (n = 26). By posttreatment, the assureds had lost significantly more weight (10 +/- 6 vs. 7 +/- 7 kg). Regrouping the data for analysis by posttreatment self-efficacy types demonstrated transitions in the self-efficacy beliefs of the women during treatment. Those who were disbelievers at baseline but became assureds posttreatment (n = 7) lost twice as much weight as the women who started and finished as disbelievers (n = 19) (10 +/- 7 kg vs. 5 +/- 5 kg). The posttreatment assureds (n = 32) lost significantly more weight than the disbelievers (n = 22) (10 +/- 6 vs. 6 +/- 5 kg), and reported better self esteem, mood, and eating patterns. Thus, assessment of intrinsic belief systems, particularly weight-control self-efficacy, may provide new directions for designing interventions that target distinctly different needs of obese women to affect greater weight loss and more positive affective states. PMID- 8729713 TI - Measuring nicotine dependence among high-risk adolescent smokers. AB - In the present study we tested our hypothesis that because of the higher prevalence and greater intensity of cigarette smoking among vocational-technical students (N = 110; 51.8% males; mean age 17 years), adolescents might demonstrate the nicotine dependence patterns comparable to those measured in a similar fashion in a group of adult smokers (N = 173; 50% males; mean age 42 years). A modified version of the Fagerstrom Tolerance Questionnaire (FTQ) utilized in the adolescent sample was coded to make it comparable to the original FTQ used in the adult sample. The tests of item structure and internal consistency of the modified FTQ for adolescents were satisfactory; the overall mean FTQ score correlated significantly with the intensity and duration of smoking. Although the FTQ values were generally lower in the adolescent sample, 20% of students had an overall FTQ score of 6 and above, indicating substantial nicotine dependence (compared to 49% in adults). Reasons for failure of the existing adolescent smoking cessation programs as well as the rationale for adding a nicotine replacement option to the behavioral smoking cessation treatment for a subset of high-risk nicotine-dependent adolescents are discussed. PMID- 8729714 TI - Self-examination therapy as an adjunct treatment for depressive symptoms in substance abusing patients. AB - To determine the efficacy of Self-Examination Therapy as an adjunct treatment for depressive symptoms in substance abusing patients, 28 adult male volunteers from a substance abuse unit at a VA Medical Center were randomly assigned to either Self-Examination Therapy or a Current Events comparison group. Analyses indicated that participants in Self-Examination Therapy showed significantly greater improvement in depressive symptoms and overall psychopathology than participants in the Current Events comparison group. These results suggest that Self Examination Therapy is a viable adjunct treatment for substance abusers, particularly those with depressive symptoms. PMID- 8729715 TI - Adult smokeless tobacco use and age of onset. AB - To determine if an earlier age of onset is associated with greater levels of adult smokeless tobacco (SLT) use and to evaluate if this relationship differs by SLT type, 345 current adult SLT users were interviewed to obtain the age at first onset and current frequency of SLT use. The results indicated that more than twice as many SLT users began during adulthood than during preadolescence. Although preadolescent onset was associated with greater chewing tobacco use than adolescent onset, the greatest levels of both chewing tobacco and snuff use were associated with adult onset, suggesting an increased need for SLT prevention programs for young adults. PMID- 8729716 TI - The effects of cigarette smoking and alcohol consumption on blood lipids: a dose related study on men. AB - The separate and joint effects of cigarette smoking and alcohol consumption on serum concentrations of total cholesterol, high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C) and triglycerides were investigated in 46 750 men attending the BUPA Health Screening Centre in London during the period 1983-1987, after allowing for differences in age, body mass index and exercise level. Drinking alcohol was found to raise both total cholesterol and HDL-C concentrations, in such a way that HDL-C as a percentage of total cholesterol increased with increasing alcohol consumption. LDL-C concentrations increased with increasing alcohol consumption in non-smokers, but decreased in those smoking over 10 cigarettes per day. Drinking had no significant effect on triglycerides except at high levels of consumption. Smoking raised total cholesterol, LDL-C and triglycerides, but lowered HDL-C concentrations. In particular, smoking even small amounts could negate any protective benefit in HDL-C concentrations gained from moderate consumption of alcohol. Assuming a desirable lipid profile to consist of low total cholesterol, LDL-C and triglycerides and a high HDL-C value, this is best achieved for men by being a non-smoking moderate drinker. PMID- 8729717 TI - Differences in serum luteinizing hormone measurements by immunoradiometric assay induced by kinetic manipulation of assay conditions are dependent on the endocrine milieu of serum. AB - Divergent estimates for luteinizing hormone (LH) in individual serum samples may be given by different immunoassays. In order to investigate this phenomenom further, we have studied the effect of differences in assay kinetics within the same immunoradiometric assay (IRMA) configuration on LH measurement in sera from different endocrine states. Three pairs of monoclonal/polyclonal two-site IRMA systems for LH were developed from three LH monoclonal antibodies and a common polyclonal anti-human chorionic gonadotrophin. For IRMA systems a short and long assay, which were different only with respect to the incubation time (1/2 h and overnight respectively), of the labelled monoclonal first antibody were performed. The IRMAs were all standardized against the LH international reference preparation 68/40. LH concentrations were measured by all the IRMAs in sera obtained from normal men (n = 11) and from women with polycystic ovarian syndrome (PCO; n = 13). In normal men, there were no differences in LH estimates between the short and the long assays of the three IRMA systems, and the ratios of long to short assays were similar for all the systems. However, in PCO there were significant differences between short and long assays and the ratios of long to short assays were different for the IRMA systems. These results indicate that kinetic differences between IRMAs of the same antibody configuration can be associated with differences in measured LH concentrations, depending on the endocrine status of the sera studied. As LH glycoform patterns are known to differ between normal men and PCO, the observed changes in LH estimates may be due to the different glycoform composition. PMID- 8729718 TI - The use of the short tetracosactrin test for the investigation of suspected pituitary hypofunction. AB - The short Synacthen (tetracosactrin) test (SST) is an established method of assessing adrenocortical reserve, and is increasingly replacing the insulin tolerance test (ITT) for the assessment of the hypothalamic-pituitary-adrenal (HPA) axis. However, there is no consensus on how the test is performed, and various time points and routes of administration are used. The present study was done, first, to determine whether there was any difference in cortisol responses when Synacthen was administered intramuscularly compared with intravenously and, secondly, to compare cortisol responses at 30 and 60 min. We found no significant difference between the two routes of administration. However, cortisol responses at 60 min were significantly higher than at 30 min (P < 0.05). Previous validations for the use of the SST in place of the ITT have used cortisol responses 30 min after Synacthen. We conclude that where the SST is used to assess the HPA axis, cortisol response at 30 min after intravenous Synacthen should be used. PMID- 8729719 TI - Indirect sandwich enzyme-linked immunosorbent assay (ELISA) for plasma apolipoprotein E. AB - We have developed an indirect sandwich ELISA for measuring plasma apolipoprotein E (apo-E), using commercially available antibodies. A monoclonal anti-apo-E was used as the capture antibody and the captured apo-E detected with polyclonal anti apo-E antiserum (goat). The detecting antibody was quantitated using horseradish peroxidase-conjugated rabbit immunoglobulin to goat immunoglobulins. There was no detectable cross-reactivity between the three antisera. Interference with the assay by apolipoprotein A-1, bilirubin and haemoglobin was not significant up to 1.7 g/L, 1250 mumol/L and 13.0 g/dL, respectively. The ELISA method showed high correlation with an established immunonephelometric method (ELISA apo.E = 1.04 Immunonephelometric apo-E + 16; r2 = 0.954, P < 0.0001, n = 39). The assay has a measuring range between 5 and 560 mg/L. The coefficient of duplicates was 2.0%, within-run coefficients of variation (CV) ranged from 3.7 to 6.0% and between-run CV from 6.1 to 15.1%. The reference range determined for 168 normotriglyceridaemic subjects was 20 to 130 mg/L. In an analysis of the lipoprotein subfractions isolated by ultracentrifugation as the fraction of density less than 1.25 g/mL and separated by gel permeation chromatography, apo-E was found to be associated with very low-density lipoprotein and large high density lipoprotein. PMID- 8729720 TI - Measurement of bone-specific alkaline phosphatase by an immunoselective enzyme assay method. AB - We evaluated a new immunoselective enzyme assay of bone-specific alkaline phosphatase (ALP). The monoclonal antibody used in this assay was raised against purified bone-specific ALP obtained from SAOS-2 human osteosarcoma cell line. Calibration was based on the enzyme's own activity. The relative activity of the antibody was 100% with bone ALP, 8.7% with liver ALP, and 0% with placental and intestinal ALPs. Intra- and inter-assay coefficients of variation were less than 4%. The sensitivity of the assay was 0.7 U/L, and the linearity extended from 2 to 140 U/L. The recovery of bone-specific ALP standard added to serum was 94 106%. The correlation coefficient between this method and the polyacrylamide gel (PAG) electrophoretic method was 0.94. The mean value of bone-specific ALP in 89 healthy adults (mean age 29 years, SD 5 years) was 18.5 U/L (SD 4.1 U/L). Interestingly, mean bone-specific ALP activities in 60 premenopausal women (mean age 39 years, SD 8 years) and 70 postmenopausal women (mean age 57 years, SD 5 years) were 20.3 U/L (SD 6.5 U/L) and 31.1 U/L (SD 11.1 U/L), respectively. The age-related increase in bone-specific ALP was significant and more pronounced in women (P < 0.01). We conclude that this new immunoassay of bone-specific ALP would be useful for clinical investigation of patients with osteoporosis or other metabolic diseases of bone. PMID- 8729721 TI - Detection of Bence-Jones protein in serum by immunoblotting. AB - To detect Bence-Jones protein (BJP) in serum we precipitated intact immunoglobulins (Ig) using polyethylene glycol (PEG) and subjected the BJP in solution to electrophoresis in agarose gel, followed by transfer to a polyvinylidene difluoride membrane, and immunoenzymatic staining (successively using rabbit anti-human light/heavy chain of Ig, biotinylated swine anti-rabbit Ig, and alkaline phosphatase-conjugated streptavidin). Treatment with PEG effectively reduced background staining of polyclonal Ig in the immunoblots, although intact monoclonal Ig was not always completely removed. To compare the present method with immunoelectrophoresis (IEP), we selected samples from patients demonstrating BJP by IEP in both serum and urine (n = 40), serum only (n = 18), and urine only (n = 32); 21 of these patients had BJP alone and 69 had BJP in addition to intact monoclonal Ig. Efficiency of detection of BJP in serum was increased by the present method: serum BJP was detected in 70 patients by the present method versus 58 by IEP. The present method demonstrated single BJP bands in the samples from 16 patients (kappa, n = 7; lambda, n = 9) and multiple BJP bands (range: 2-9) in the samples from 54 patients (kappa, n = 31; lambda, n = 23). This method could be useful for detecting BJP in serum from patients suspected of having light chain gammopathy (without the need for urine testing) and may complement urine testing in patients excreting polyclonal free light chains of Ig. PMID- 8729722 TI - A practical approach to minimizing inaccuracy through traceability, using limited resources. PMID- 8729723 TI - Apolipoprotein E and epilepsy. PMID- 8729724 TI - Apolipoprotein E intrathecal synthesis is decreased in multiple sclerosis patients. PMID- 8729725 TI - Clinically significant interference by haemoglobin F during pregnancy with the measurement of HbA1 using the Corning agar electroendosmotic technique. PMID- 8729726 TI - An external quality assessment scheme for amiodarone and desethylamiodarone: initial results. PMID- 8729727 TI - Progression of patients with a borderline raised thyroid-stimulating hormone concentration. PMID- 8729728 TI - Paradoxical high-density lipoprotein reduction induced by fenofibrate and ciprofibrate. PMID- 8729730 TI - Massive hypercortisolaemia secondary to stress. PMID- 8729729 TI - Interference by biotin therapy on measurement of TSH and FT4 by enzyme immunoassay on Boehringer Mannheim ES700 analyser. PMID- 8729731 TI - A case of creatine kinase non-M activity in human plasma. PMID- 8729732 TI - Haemochromatosis associated with arthritis and hypopituitarism. PMID- 8729733 TI - Anticoagulant effects of antioxidant capacity. PMID- 8729734 TI - Origins of the sex difference in human urinary free cortisol excretion. PMID- 8729735 TI - Interpretation of cortisol response to synacthen. PMID- 8729736 TI - Structural basis for oxygen delivery: muscle capillaries and manifolds in tuna red muscle. AB - We summarize our morphometric data on fiber vascularization and aerobic capacity in red muscle of tuna (Katsuwonus pelamis), compared to intensely aerobic flight muscles of hummingbird (Selasphorus rufus, BW 3-4 g) and bat (Eptesius fuscus, BW 15-16 g, Pipistrellus hesperus, BW 3-5 g). Three characteristic features of high flux paths for oxygen: (a) small fiber size, (b) dense capillary network and (c) high mitochondrial volume density were found in tuna, but they were not as pronounced as in hummingbird and bat flight muscles. A particular arrangement of capillary manifolds, also seen in flight muscle of birds but not in bats, was found in tuna, forming dense envelopes of capillary branches around portions of muscle fibers. However, all indexes of fiber capillarization were relatively low in tuna red muscle for its mitochondrial volume, compared with other intensely aerobic muscles. Capillary length per unit volume of mitochondria, and capillary surface per mitochondrial inner (and outer) membrane surface area, were about one half of those in hummingbird or bat flight muscles. Consistent differences exist in the size of the capillary network for the size of the mitochondrial compartment in highly aerobic red muscle of tuna compared with bird and mammal. PMID- 8729737 TI - Bibliography of Comparative Biochemistry and Physiology A. Generated from the Current Awareness in Biological Sciences database. PMID- 8729738 TI - Methodologies to assess moderate therapeutic effects. PMID- 8729739 TI - The treatment of allergic rhinitis improves the recovery from asthma and upper respiratory infections. AB - Forty-six asthmatic children with repeated respiratory infections presented symptoms of allergic rhinitis. All patients were treated locally for allergic rhinitis either with disodium cromoglycate or beclomethasone dipropionate. After six months of treatment, 95% of the children showed improvement of allergic rhinitis and 84% improvement of bronchial asthma, as well as fewer infections. We concluded that allergic rhinitis plays an important role in facilitating infections of the upper respiratory tract, and a possible association of rhinitis, viral infections and bronchial asthma is discussed. PMID- 8729740 TI - Transdermal glyceryl trinitrate (nitroglycerin) in healthy persons: acute effects on skin temperature and hemodynamic orthostatic response. AB - In order to find an explanation for individual reactions to transdermal glyceryl trinitrate (GTN) we studied the skin temperature and hemodynamic reactions in 63 healthy persons. The data were obtained before and after the application of GTN and Glycerin (GL) placebo patches, during one hour. The skin temperature was measured on both forearms, the local (left sided) and systemic (right sided) reaction on GTN was related to the skin fold and the calculated body fat content. The bilateral rise of skin temperature and its duration was higher and longer in obese than in lean persons mainly in obese women. The UV induced thermo and the later photothermoreaction (Erythema) was reduced on the left forearm after the application of GTN and GL patches. The observed hemodynamic GTN effect confirmed known postural reactions, such as decreased arterial pressure (delta mAP = 2.9%), increased heart rate (delta HR = +7.4%) and QTc prolongation (delta QTc = +4.9%) in upright position. An adverse drug effect with increased mean blood pressure (delta mAP = +12%) and increased heart rate (delta HR = +10.4%) mainly in supine position was observed in 11% of the participants, but only in men. Such a reaction was already described by Murell, 1879. Individual GTN effects were analyzed and related to habits and family history. In male smokers and in persons with hypertensive and diabetic close relatives, the hypotensive GTN effect was accentuated in supine position. In the upright position the group with hypertensives in the family presented a moderate hypotensive reaction without secondary tachycardia and the smokers presented only a slightly increased heart rate. Our observations suggest that individual reactions to transdermal glyceryl trinitrate (GTN) with its active component nitric oxide (NO) depends on physiological conditions, related to endogenous vasoactive substances, mainly the interaction with EDRF (the endogenous NO) and the activity of the Renin Angiotensin System. PMID- 8729741 TI - The value of therapeutic planning and the degree of experience of the surgical team on the results of cancer treatment of the larynx and hypopharynx. AB - This is a prospective study supported by 170 cases of epidermoid carcinoma of the larynx or hypopharynx, treated during the period from January of 1981 to January of 1988, at the Head and Neck Surgery Service of the Heliopolis Hospital Complex, Sao Paulo. The objective of this project was to evaluate the importance of surgeon experience with regard to the rates of post-operative complications and the rates of relapse and survival. The results of the 8 surgical specialists who integrate the permanent staff at the institute and who different varying degrees of experience with regard to time spent exercising their specialties were compared. The results obtained did not show a significant difference among the various surgeons, and this uniformity is explained by the fact that all the therapeutic planning was elaborated through consensus of the whole group, and this could have minimized the effect of experience of a surgeon responsible for the operation. The authors emphasize the importance of pre-operative evaluation for good results and propose that it is in the direction of complete mastery of preliminary work in the area that programs for the formation of new specialists should be directed. PMID- 8729742 TI - Postmenopausal genital bleeding. AB - The study was conducted on 748 women who reported genital bleeding occurring at least one year after the last menstruation. Benign causes were most frequent than malignant causes. Among the benign causes, the most frequent were cervicitis (19.95%), prolapsed uterus with decubitus ulcer (19.41%), dysfunctional hemorrhage (13.29%) and endometrial polyps (12.77%). In the group of malignant causes, cancer of the cervix was the neoplasm most often detected (59.26%); endometrial cancer was next, affecting 29.63%. The cancer of the cervix/cancer of the body ratio was 2:1. In summary, many causes, both benign and malignant, can provoke abnormal postmenopausal bleeding. Thus, curettage of the uterus should be reserved for doubtful cases, i.e., in situations in which, after all non invasive methods of investigations have been exhausted, the possibility of the occurrence of malignant lesions still persist. PMID- 8729743 TI - Leiomyoma of the breast. A case report. AB - A rare case of leiomyoma of the breast is reported, with a discussion of the clinical aspects and of the differential diagnosis. Excluding tumors originating from the areolar-papillary complex and the skin, this neoplasm is extremely rare, with only 11 cases reported so far. The histogenesis of the lesion is still controversial. PMID- 8729744 TI - Progressive muscular dystrophy--Duchenne type. Controversies of the kinesitherapy treatment. AB - The authors carried out a study of children with progressive muscular dystrophy of Duchenne type (DMD), giving special attention to physiatrical follow-up, having in mind that the practice of exercises has been debated very much in the specialized literature. The goal of this study is to try to settle the limits for the utilization of kinesitherapy which should be applied only in specific situations, such as: after skeletal muscular trauma or when the respiratory system is at risk. In this situation the physiatrical procedure would be to restrict physical activity, with early use of wheelchairs and the exclusion of the use of orthoses for orthostatism. DMD, at present, has been considered a result of duplication (60%), deletion (5 to 6%) or point mutations at gen Xp21 (Zatz, 1994), that codifies a protein called Dystrophin (Hoffman et al., 1987). Dystrophin is a cytoskeletal sarcolemmic protein that constitutes about .002% of the total protein of the muscle, present in skeletal fibers concentrated in muscle tendinous joints, which supplies mechanical reinforcement to the surface of the membrane during stretching and shortening physical activity. This protein is absent in DMD cases, wherefore, the sarcolemma undergoes a segmentary necrosis losing its contractile property during eccentric and concentric physical activity. The importance of physiatrical follow-up for DMD patients is to avoid deformities and tendon shortening, to ameliorate the patient's quality of life, to provide respiratory assistance and general counseling to members of the patient's family. The objective of this study is to try to clarify the risks and possibilities of kinesitherapy applied to DMD cases. PMID- 8729745 TI - Reparation of peripheral nerves with fibrin glue prepared from snake venom. Preliminary results. AB - A new fibrin glue obtained from snake venom is presented, with possible utilization in various fields of medicine. The preparation procedures and tests in the reparation of peripheral nerves are described. The preliminary results were similar to the conventional fibrin glue procedure. PMID- 8729746 TI - Mycobacterium avium complex (MAC) isolated from AIDS patients and the criteria required for its implication in disease. AB - Before the AIDS pandemia, the Mycobacterium avium complex (MAC) was responsible in most cases for the pneumopathies that attack patients with basic chronic pulmonary diseases such as emphysema and chronic bronchitis. In 1981, with the advent of the acquired immunodeficiency syndrome (AIDS), MAC started to represent one of the most frequent bacterial diseases among AIDS patients, with the disseminated form of the disease being the major clinical manifestation of the infection. Between January 1989 and February 1991, the Section of Mycobacteria of the Adolfo Lutz Institute, Sao Paulo, isolated MAC from 103 patients by culturing different sterile and no-sterile processed specimens collected from 2304 patients seen at the AIDS Reference and Training Center and/or Emilio Ribas Infectology Institute. Disseminated disease was diagnosed in 29 of those patients on the basis of MAC isolation from blood and/or bone marrow aspirate. The other 74 patients were divided into categories highly (5), moderately (26) and little suggestive of disease (43) according to the criteria of DAVIDSON (1989). The various criteria for MAC isolation from sterile and non-sterile specimens are discussed. PMID- 8729747 TI - Detection of Cryptococcus neoformans capsular polysaccharide antigen in asymptomatic HIV-infected patients. AB - Serum samples from 242 HIV-positive persons were studied for the detection of capsular polysaccharide antigen of Cryptococcus neoformans; 193 of these patients presented less than 300 CD4+ cells/microliters of blood and 49 patients had more than 300 CD4+ cells/microliters. None of them had symptoms or signs characteristic of cryptococcosis. The capsular antigen of C. neofarmans was detected by latex agglutination technique with pronase pretreatment (IMMY, Crypto Latex Antigen Detection System, Immunomycologics Inc., OK, USA); in 61% of the samples, ELISA technique was also used (Premier, Cryptococcal Antigen, Meridian Diagnostic Inc., Cincinnati, Oh, USA). The comparative study of both methods showed that the results obtained were similar in 96.9% of the cases. The capsular antigen was detected in 13 out of 193 (6.7%) patients with less than 300 CD4+ cells/microliters. Cryptococcosis was confirmed mycologically in 3 of these 13 cases (23%) by the isolation of C. neoformans in CSF or blood cultures. Three patients, who had presented negative results of both tests for capsular antigen, suffered disseminated cryptococcosis 4 to 8 months later. The predictive diagnostic value of capsular antigen detection of C. neoformans seems to be low and we believe that it should not be done routinely in asymptomatic HIV-positive persons. PMID- 8729748 TI - Interferon gamma increases survival in murine experimental cryptococcosis. AB - Systemic disease by Cryptococcus neoformans (C. neoformans) is a common opportunistic infection in immunodeficient patients. Cellular immunity seems to be the most important determinant of resistance. The aim of this study was to assess the effect of recombinant rat interferon gamma (IFN-gamma) in murine cryptococcosis (Balb/c mice infected by IP route with the Rivas strain of C. neoformans), evaluating survival time, macroscopic and microscopic examination of the organs, and massive seeding of brain homogenate. IFN-gamma treatment, at a daily dose of 10,000 IU, did not modify significantly these variables when mice were challenged with a high inoculum (10(7) yeasts) and treatment was delayed to 5 days after infection (median survival 21 days in control mice vs. 23 days in IFN-treated). Another set of experiments suggested that IFN-gamma treatment, at a dose of 10,000 IU/day, begun at the moment of infection could be useful (it prolonged survival from 20 to 28 days, although the difference did not achieve statistical signification). When used simultaneously with infection by 3.5 x 10(5) yeasts, IFN-gamma at 10,000 IU/day for 15 days significantly prolonged survival of mice (p = 0.004). These results suggest that, depending on the experimental conditions, IFN-gamma can improve survival of mice infected with a lethal dose of C. neoformans. PMID- 8729749 TI - Fungal infections in neutropenic patients. A 8-year prospective study. AB - In this paper we report a eight-year prospective study designed to further characterize incidence, epidemiology, specific syndromes, treatment and prognosis associated with fungal infections in neutropenic patients. During the study period 30 fungal infections were diagnosed in 30 patients among 313 episodes of fever and neutropenia (10%). There were 15 cases of candidiasis, 5 pulmonary aspergillosis, 3 sinusitis by Aspergillus fumigatus, 5 infections by Fusarium sp., one infection by Trichosporon sp., and one infection due to Rhodotorula rubra. Blood cultures were positive in 18 cases (60%). The predisposing factors for fungal infection in multivariate analysis were the presence of central venous catheter (p < 0.001), longer duration of profound (< 100/mm3) neutropenia (p < 0.001), the use of corticosteroids (p < 0.001), gram-positive bacteremia (p = 0.002) and younger age (p = 0.03). In multivariate analysis only recovery of the neutropenia (p < 0.001) was associated with good prognosis whereas the diagnosis of infection by Fusarium sp. (p = 0.006) was strongly associated with a poor outcome. The death rate was 43%. There was no statistically significant difference in the death rate between patients who did receive (52%) or did not receive (50%) antifungal treatment. Identifying patients at risk, specific syndromes and prognostic factors may help to reduce the high mortality associated with disseminated fungal infections in neutropenic patients. PMID- 8729750 TI - Clinical and endoscopic findings in the mucosae of the upper respiratory and digestive tracts in post-treatment follow-up of paracoccidioidomycosis patients. AB - Systematic examination of the upper respiratory and digestive tracts (URDT) was performed in a group of 80 paracoccidioidomycosis (PCM) patients submitted to post-treatment follow-up ranging from 8 months to 17 years. Mucosae of the URDT had been involved prior to specific treatment in 74 patients, distributed as follows: oropharynx, 50 (41 alone, 7 in association with the larynx, and 2 with the nasal mucosa); larynx, 30 (23 alone and 7 in association); and nasal mucosa, 3 (1 alone and 2 in association). Inactive lesions were observed in all the 50 patients with lesions of the oropharynx, 3 of whom with deforming scars (1 with retraction of the tongue and 2 with narrowing of the oral orifice). One case presented a destructive lesion, with perfuration of the palate. Of the other 46 cases, examination showed nacreous white striated scars which were nearly imperceptible in some cases and in others displayed partial retraction of anatomical structures without any alteration of their features. Patients presented a high rate of missing teeth. In 3 patients with involvement of the nasal mucosa, none of whom presented active PCM lesions, 2 still had nasal voices. In 30 patients with lesions of the larynx, 1 suffered a relapse of PCM and 2 developed epidermoid carcinoma. Of the other 27 cases, none of whom had active PCM lesions, 15 presented dysphonia, 3 were tracheotomized, and 9 were asymptomatic. PMID- 8729751 TI - Chagas' disease in the Brazilian Amazon. III. A cross-sectional study (1). AB - Two serological surveys for Chagas' infection were carried out, in 1991 and 1993, respectively, using a conglomerate family samples from the residents in the town of Barcelos (in the northern part of the State of Amazonas, on the right bank of the Rio Negro, 490 Km up-river from Manaus), using indirect immunofluorescent tests for anti-T. cruzi antibodies. In the first survey (1991), 628 blood samples from the residents of 142 dwellings were tested, showing positive in 12.7% for anti-T. cruzi antibodies and in 1993 an other 658 samples from residents of 171 dwellings showed positive in 13.7% of the tests, thus confirming the previous results. From 170 individuals with positive serology for T. cruzi antibodies, 112 (66%) were interviewed and submitted to electrocardiographic and clinical examinations; 82 (73.2%) of them gave consent for xenodiagnosis. From the 112 interviewed 52 (46.4%) recognized the triatomines as "piacavas' lice", 48 (42.8%) knew the bugs from their work places being gatherers of piacava fibers in rural areas and 19 (16.9%) said that have been bitten by bugs in their huts. Only 2 (2.4%) of 82 xenodiagnosis applied were positive for T. cruzi and 9 (8%) of the ECG had alterations compatible with Chagas' disease. PMID- 8729752 TI - Serological findings during a measles outbreak occurring in a population with high vaccine coverage. AB - From March 1991 to April 1992, serum samples for IgM detection were collected from 112 clinical measles cases reported to the Health Department of Niteroi, State of Rio de Janeiro. The positivity exceeded 90% for specimens collected from the 5th to the 29th day after the onset of the disease. After day 30 a decline in IgM detection was observed, although positivity has been detected up to 90 days after the onset of the symptoms. Forty-four patients (48.9%) with an IgM response had a history of prior measles vaccination. In 5 of the 22 measles-IgM negative cases the infection was due to other agents (rubella: 4 cases, dengue: 1 case). These results show that sensitivity of the test employed for confirming suspected measles cases is high, even in vaccinated patients. PMID- 8729753 TI - Risk factors and prevalence of antibodies against hepatitis A virus (HAV) in children from day-care centers, in Goiania, Brazil. AB - A seroepidemiologic survey about hepatitis A virus (HAV) infection was carried out in a group comprising 310 children, ranging in age from 3 months to 9 years, from day-care centers, in Goiania, a middle sized city in the central region of Brazil. The biomarkers employed in the investigation of previous infection include total IgG and IgM anti-HAV antibodies, and for the detection of more recent infection, IgM anti-HAV antibodies were analyzed. The study was performed in 1991 and 1992. According to the results, 69.7% of the children presented total IgG/IgM anti-HAV antibodies, with 60% of the group in the age range of 1 to 3 years. Among 10 day-care centers analyzed, the prevalence of the biomarker IgM anti-HAV was 3.2%, with an uniform distribution of the cases in the group of children ranging in age from 1 to 4 years. Multivariate analysis was performed to investigate the sociodemographic factors that could influence the results. It was verified that the risk for the infection increased with the length of the attendance in the day-care centers, i.e., the risk for children with attendance of one year or more was 4.7 times higher, when compared with children with one month attendance (CI 95% 2.3-9.9). According to the results, hepatitis A is an endemic infection in day-care centers in the study area. The length of attendance in the day-care settings was demonstrated to be a risk factor for the HAV infection. Such findings suggest that if hepatitis A vaccination becomes available as a routine policy in our region, the target group should be children under one year. Moreover, those children should receive the vaccine before they start to attend the day-care centers. PMID- 8729755 TI - Schistosoma mansoni Sambon, 1907: comparative morphological studies of some Brazilian strains. AB - The morphology of Schistosoma mansoni adult male worms from three strains which have been maintained in albino mice for several generations, was compared to a strain that has been isolated from the natural host Nectomys squamipes (Rodentia:Muridae) captured in Sumidouro (Rio de Janeiro State) and have been maintained in the same sylvatic rodent under laboratory conditions. Total length of specimens, distance between suckers, the number of testes and extention of testes grouping were the taxonomic characters analysed. The worms recovered from N. squamipes showed expressive differences (p < 0.01) compared to the other strains regarding the considered morphological characters. The strains that were maintained in mice presented statistical differences (p < 0.01) in several characters. Some adult worms besides the normal position of the testes also showed an atypical arrangement of these glands. It can be concluded that the morphology of adult worms may be used to distinguish S. mansoni strains and that morphological changes in adult worms are not induced by successive inoculations of a strain in mice. PMID- 8729754 TI - Hyperimmune antirabies sera titration by standard mouse neutralization and counterimmunoelectrophoresis tests, comparing results of different laboratories. AB - To determine the rabies antibody level of twenty-four hyperimmune equine sera, Standard Mouse Neutralization (SMN) and Couterimmunoelectrophoresis (CIE) tests were carried out, both at the Instituto Butantan (IB) and Instituto Panamericano de Proteccion de Alimentos y Zoonosis (INPPAZ). Statistical analysis has shown a correlation (r) of 0.9317 between the SMN and CIE performed at the IB, while at the INPPAZ it scored 0.974. Comparison of CIE data of both laboratories yielded a correlation of 0.845. The CIE technique has shown to be a sensitive and efficient as the SMN in titrating antirabies hyperimmune equine sera. Based on CIE results, a simple, rapid and inexpensive technique, titers of sera antibody can be rellably estimated in SMN test. PMID- 8729756 TI - The value of adenosine deaminase (ADA) determination in the diagnosis of tuberculous ascites. AB - In order to evaluate the role of the determination of adenosine deaminase activity (ADA) in ascitic fluid for the diagnosis of tuberculosis, 44 patients were studied. Based on biochemical, cytological, histopathological and microbiological tests, the patients were divided into 5 groups: G1-tuberculous ascites (n = 8); G2-malignant ascites (n = 13); G3-spontaneous bacterial peritonitis (n = 6); G4-pancreatic ascites (n = 2); G5-miscelaneous ascites (n = 15). ADA concentration were significantly higher in G1 (133.50 +/- 24.74 U/l) compared to the other groups (G2 = 41.85 +/- 52.07 U/l; G3 = 10.63 +/- 5.87 U/l; G4 = 18.00 +/- 7.07 U/l; G5 = 11.23 +/- 7.66 U/l). At a cut-off value of > 31 U/l, the sensitivity, specificity and positive and negative predictive values were 100%, 92%, 72% and 100%, respectively. ADA concentrations as high as in tuberculous ascites were only found in two malignant ascites caused by lymphoma. We conclude that ADA determination in ascitic fluid is a useful and reliable screening test for diagnosing tuberculous ascites. Values of ADA higher than 31 U/l indicate more invasive methods to confirm the diagnosis of tuberculosis. PMID- 8729757 TI - Autoantibodies before, during and after administration of recombinant interferon alpha for chronic viral hepatitis. AB - This study was undertaken to investigate the presence of autoantibodies in patients with chronic viral hepatitis B and C, before, during and after interferon-alpha (IFN-alpha) therapy and to study their relation to dose and type of IFN-alpha and response to treatment. Fifty patients with chronic hepatitis were divided in two groups, a control-group of 21 patients (10 type B and 11 type C) who were followed for 6 months without treatment and an IFN-group consisting of 29 patients (8 type B and 21 type C) who received IFN therapy for 6 months. Serum samples were tested for a range of antibodies at the start of the study, during therapy and at the end of the 6 month period. Antibodies tested for included: antinuclear, smooth muscle, antimitochondrial, parietal cell and thyroid microsomal. Four (8%) of the total patient group had autoantibodies at the beginning of the study (two in each group). During the follow-up period no patient in the control group developed antibodies compared with 3 (11%) patients in the treatment group. Autoantibodies developed in patients treated with higher doses of IFN and were found in those patients who tended to show a poor response to IFN-therapy. Further studies are needed to establish the relationship between poor response to IFN-alpha and development of autoantibodies. PMID- 8729758 TI - Is CK-MB isoenzyme useful for diagnosis of cardiac involvement in icteric leptospirosis? AB - In the absence of heart failure or cardiogenic shock, cardiac involvement diagnosis in icteric leptospirosis is possible on the basis of abnormal electrocardiograms. As metabolic and electrolytic disorders are frequently seen during acute leptospirosis infection, they may be responsible for some electrocardiograms changes. We conducted a study to assess if creatine phosphokinase isoenzyme determinations are useful in selecting patients with a high cardiac involvement suspicion. Sixty-nine patients were studied prospectively. Ten patients out of 16 with cardiac involvement and 25 without had high CK-MB levels (p > 0.05), although mean values of abnormal CK-MB levels were higher in the group with cardiac involvement (p < 0.05). Our analysis indicates that serum CK-MB determination does not provide a specific indicator of myocardial involvement in the course of icteric leptospirosis. PMID- 8729759 TI - The relevance of laboratory diagnosis of human cryptosporidiosis and other coccidia. AB - Human infection by Cryptosporidium spp and other coccidia are due to opportunist non-host specific microorganisms. In HIV seropositive patients, the gastrointestinal symptoms accompanying such infections may be serious and prolonged and may include nausea, low-grade fever, abdominal cramps, anorexia and watery diarrhoea. We studied 188 stool samples from 111 patients (84 men and 27 women) with diarrhoea. A modified Ziehl-Nielsen technique for the detection of Cryptosporidium spp and Isospora belli was employed. The mean age of the patients was 31 years. Cryptosporidium spp was seen in 18% (n = 20) of the patients, 90% (n = 18) of whom were HIV seropositive. Isospora belli was recorded only from HIV seropositive patients (5.4% of all the patients studied and 6.5% of those who were HIV seropositive). These data confirm the good results obtained with this technique for the identification of Cryptosporidium spp and other coccidia and also reaffirm the clinical importance of correctly diagnosing the cause of diarrhoea, particularly in HIV seropositive patients. PMID- 8729760 TI - Parasitological diagnosis of schistosomiasis mansoni: concomitant utilization of Kato-Katz method and hatching test. PMID- 8729761 TI - Size of Cryptosporidium oocysts excreted by symptomatic children of Santiago, Chile. PMID- 8729763 TI - [Occipital epileptiform activity with or without blockade with eye opening: a comparative clinico-electroencephalographic study]. AB - Occipital epileptiform activity blocked by eyes opening (OEAB) is primarily observed in childhood epilepsy with occipital paroxysms but may be also found in cases of benign childhood epilepsy with centrotemporal spikes, cryptogenic/symptomatic partial epilepsies, idiopathic generalized epilepsies and in patients with no seizures. We studied comparatively EEG and clinical features of 20 patients with OEAB and 120 patients with occipital epileptiform activity not suppressed by eyes opening (OEA). Epileptiform activity (OEA or OEAB) was registered until the 7 years of age in 67.5% of the patients with OEA and in 45% of the patients with OEAB. Epileptic seizures began before 4 years of age in 44.4% of the cases with OEA and in 14.2% in the group OEAB. Febrile convulsions alone occurred only in the OEA group (12.5%). Developmental retardation was referred in 27 patients with OEP. The occurrence of spike and wave complexes was statistically more frequent among patients with OEAB (80.0%) as opposed to patients with OEA (17.5%). Our findings suggest clinical and electroencephalographic differences between children with OEA and OEAB. PMID- 8729762 TI - Hepatic enzymes' level during chronic use of anticonvulsant drugs. AB - We studied retrospectively 894 adult epileptic patients treated during the period from 1983 to 1992. Hepatic enzymes abnormal values were seen in 49% (n = 438). In 22.3% (n = 200), at least 2 enzyme levels in different moments were altered. They were divided in three groups: GI with alterations at transaminases (3%, n = 6), GII with alterations at GGT and AP enzymes (72%, n = 144) and GIII with alterations in both groups (25%, n = 50). No patient developed clinical symptoms of liver disease. The increase of gamma-glutamil-transferase (GGT) and alkaline phosphatase (AP) levels is frequent and not necessarily pathological. Slight increase of transaminases can occur with no clinical correlation. The routine screening of hepatic enzymes level during the chronic use of anticonvulsivant drugs in adults has a questionable value. PMID- 8729764 TI - [Comparison between HDL and LDL cholesterol as risk factors for carotid atherosclerosis]. AB - In order to find out whether there exists a relationship between HDL and LDL serum levels and atherosclerotic plaques in the carotid artery (CA), a prospective study was carried out involving 125 patients. They were aleatorily included, of both sexes and age between 45-75 years old. HDL and LDL serum levels were measured as well CA ultrasonographic mode B investigated. It was verified that the number of patients having atherosclerotic plaques in CA was inversely proportional to the HDL levels. We came up with the same ratio when the HDL levels were compared to the number of patients having CA stenosis. These results were statistically significant (x2 = 6.57 and x2 = 9.24 respectively; critical x2 = 5.99 to alpha = 5%). A direct ratio was also found out in the relationship between the presence of atherosclerotic plaques in the CA and the LDL serum levels (greater in patients with plaques). However no relationship between LDL levels and the occurrence or not of CA stenosis in patients having plaques was demonstrated. The results were not statistically significant (x2 = 0.97 and x2 = 0.41, respectively, critical x2 = 5.99 to alpha = 5%). The obtained results seem to be at least in part in agreement with literature findings. Statistically significative results in the comparison of HDL and LDL levels with the grade of stenosis of the CA were not found out (x2 = 11.78 and x2 = 4.03, respectively; critical x2 = 12.59 to alpha = 5%). PMID- 8729765 TI - [Levodopa-induced dyskinesia in 176 patients with Parkinson's disease]. AB - Dyskinesias are frequently observed in parkinsonian patients during levodopa treatment. The occurrence of these movement disorders usually makes the therapeutic management of the patients very difficult. The clinical characteristics of 176 patients with dyskinesias were retrospectively studied. Dyskinesias occurred, on average, after 6.2 years of duration of Parkinson's disease and after 4.2 years on treatment with levodopa. Patients were more likely to have dyskinesias during more advanced stages (measured by Hoehn and Yahr scale). Peak of dose and square wave were the types of dyskinesia more frequently described and were associated with choreic movements in most cases. Dystonia occurred in 40% of the cases and was predominant in end of dose and diphasic dyskinesias. Thirty-five percent of dystonia cases presented as "early morning dystonia". Chorea was the most frequent involuntary movement and mostly generalized. Dystonia was most commonly described in lower limbs. Orofacial dyskinesia, when occurred alone, was more frequently seen in old rather than young patients. When dyskinesia was unilateral it was more likely to occur in the side where Parkinson's disease was more severe. PMID- 8729767 TI - [Longitudinal psychological study in myotonic dystrophy]. AB - The authors studied in two occasions a group of 12 patients with myotonic dystrophy in a mean interval of three years and a half between the examinations. The neuro-psychological battery included the following tests: Raven's progressive matrices (coloured and general scales), Wechsler children intelligence scale (WISC), Kohs' blocks and Piaget-Head. 50% of the patients had better scores on the second examination on RCPM, 81.89% on WISC-digit span, 63.67% on WISC numbers, 44.44% on Piaget-Head 2 and 60% on Kohs' blocks. However, on Piaget-Head 1-3, the majority had worse results (87.56%) with statistical significative difference (p < 0.05). Though the scores from Kohs' blocks were better in 60% of the patients with p < 0.05, we have to consider that 60% had 0 point on first examination being their scores a little better on second one. This is enough to result on statistical significative difference, however, very low if compared to normal subjects. PMID- 8729766 TI - [Abnormalities of pattern reversal evoked visual potential in patients with defined multiple sclerosis]. AB - The pattern reversal visual evoked potential with checks of 14' and 28' was retrospectivelly studied in 28 patients with definite multiple sclerosis. We observed abnormal responses in 27/28 (96.4%) patients, in 31/36 (86%) of symptomatic eyes, and in 16/20 (80%) of asymptomatic eyes. When we classified the abnormalities in each eye according to the findings obtained with each check, there was a correlation between the pattern of abnormalities and the severity of visual involvement. Occasionally there were isolated abnormalities of N75 or only in P100 obtained with 28' checks. In conclusion the methodology applied was very sensible in detecting abnormalities in visual pathway. We could classify the findings in each eye and correlate them with the severity of visual involvement. The findings showed uneven distribution of lesions in visual pathway, affecting preferentially the central vision afferents. PMID- 8729768 TI - [Adenosine deaminase in the cerebrospinal fluid of patients with acquired immunodeficiency syndrome]. AB - The adenosine-deaminase (ADA) activity was evaluated in CSF samples from 263 patients with AIDS. An elevated ADA activity in CSF was found in patients with: antibodies to toxoplasmosis, syphilis or cytomegalovirus; Cryptococcus neoformans or their antigens; tuberculous meningitis; lymphoma. There was no statistical difference among all these groups in respect to ADA activity. However, the ADA activity in CSF from AIDS patients without CSF changes other than HIV antibodies, even unspecific changes, was not elevated. This may suggest that ADA is related to AIDS associated pathologies activity rather than to HIV infection itself. PMID- 8729770 TI - Pulmonary involvement in patients with HTLV-I associated myelopathy. AB - Pulmonary involvement in HTLV-I patients has been identified by several authors in Africa, Japan and Brazil. The objective of this controlled study is to establish an association between HAM and lymphocytosis in the bronchoalveolar fluid (BAL). Thirty-five adult patients with non-traumatic and non-tumoral myelopathy filled out a detailed historical survey and underwent a neurological examination, a thoracic radiological evaluation and a CSF examination. Of the patients in this sample, 22 were diagnosed with HTLV-I associated myelopathy and 13 had myelopathies with other etiologies. Lymphocytosis in the BAL fluid was detected in 18 (82%) of the HAM patients and in 2 (15%) of non-HAM patients. We concluded that the lung represents an important organ in the pathogenesis of HAM. PMID- 8729769 TI - The value of CSF analysis for the differential diagnosis of HTLV-I associated myelopathy and multiple sclerosis. AB - Cerebrospinal fluid (CSF) and serum of 17 patients with HAM/TSP (HTLV-I associated myelopathy/tropical spastic paraparesis), six with multiple sclerosis and six with idiopathic epilepsy (non inflammatory control) from Brazil were analysed for the presence of intrathecal synthesis of virus-specific antibodies against measles, rubella, varicella zoster virus and herpes simplex virus by enzyme-linked immunosorbent assay (ELISA). All HAM/TSP and multiple sclerosis cases had an intrathecal immune response (oligoclonal IgG). In HAM/TSP, only 1/17 case showed a polyspecific intrathecal immune response against measles and rubella virus. In multiple sclerosis, specific antibodies against measles and rubella (MRZ response) were observed in all patients but not in the control with idiopathic epilepsy. The diagnostic and theoretical relevance of mono- and polyspecific immune responses is discussed for these chronic neurological diseases. PMID- 8729771 TI - HTLV-I associated myelopathy in Porto Alegre (Southern Brazil). AB - HTLV-I associated myelopathy/tropical spastic paraparesis (TSP/HAM) have been increasingly described in practically all regions of Brazil. Five confirmed and documented cases of patients with TSP/HAM in Rio Grande do Sul are reported; in all of them spastic paraparesis, neurogenic bladder and superficial and/or profound sensitive disorders were observed in variable degrees. One patient presented a relapsing-remitting course with a cerebellar ataxia (multiple sclerosis-like pattern). Everyone was submitted to clinical, serological, urodynamic, neurophysiologic and neuroradiologic investigation. The aim of this study was to present the southern region of Brazil as an area with significant endemicity for HTLV-I/II infection (prevalence of 0.42% between blood donors), and also to show the existence of patients with neurologic disease associated with this retrovirus. PMID- 8729772 TI - HTLV-I associated uveitis, myelopathy, rheumatoid arthritis and Sjogren's syndrome. AB - A 62 year-old white female presented with a 10-year-history of slowly progressive spastic paraparesis, pain and dysesthesia in the lower limbs and sphincter disturbance. A few years after the onset of the neurologic symptoms she developped migratory arthritis with swelling of the knees and pain on palpation of knees and fingers, dry eyes, mouth and skin. Two months before admission she presented bilateral nongranulomatous anterior uveitis. Examination revealed spastic paraparesis with bilateral Babinski sign, a decreased sensation level below L3, decreased vibration sense in the lower extremities, and a postural tremor of the upper limbs. Laboratory work-up disclosed HTLV-I positive tests in the blood and cerebrospinal fluid (CSF), and a mild pleocytosis in the CSF with a normal protein content. Nerve conduction velocity studies were normal. The present case shows the association of uveitis, arthritis and Sjogren's syndrome in a patient with tropical spastic paraparesis/human T-cell lymphotropic virus type I (HTLV-I) associated myelopathy (TSP/HAM), and illustrates the wide spectrum of clinical manifestations which may accompany this infection with this virus. PMID- 8729773 TI - Von Hippel-Lindau's disease: report of three cases and review of the literature. AB - The authors present the autopsy findings of two related patients and the biopsy findings of a third member of the family. The oldest member was 34 years old at death and on postmortem examination he had haemangioblastomas in the retina, cerebellum, medulla and spinal cord. Other findings were renal cell carcinoma, phaechromocytoma, cysts of kidney and pancreas, hydromyelia and atypical meningiomas. His brother died when 30 years old. The autopsy revealed haemangioblastomas of cerebellum, renal cell carcinoma and a clear cell cystadenoma of epididymus. The third patient was the daughter of the first and presented with headache and dizziness. CT-scan showed a cerebellar haemangioblastoma. Epidemiological considerations on the commonest visceral and CNS lesions and a review of current diagnostic criteria are discussed. PMID- 8729774 TI - [Brown-Vialleto-van Laere syndrome: report of 2 cases]. AB - Brown-Vialetto-van Laere syndrome is a rare hereditary or sporadic degenerative disorder characterised by progressive sensoryneural deafness, followed or accompanied by cranial nerve palsies. The anterior horn cells and the optic pathways may be involved in some cases. We report two cases, and comment the differential diagnosis and the relationships of this syndrome to the muscle spinal atrophies and the hereditary deafness. PMID- 8729775 TI - Myoclonic epilepsy of late onset in trisomy 21. AB - We report the case of a patient with trisomy 21 (T21) with late onset epilepsy. The electroclinical features were of myoclonic jerks on awakening and generalised tonic clonic seizures, with generalised spike and wave on EEG, and a progressive dementia. As familial Alzheimer's dementia and progressive myoclonic epilepsy (Unverricht-Lundborg type) are both linked to the chromosome 21, this case may represent a distinct progressive myoclonic epilepsy related to T21. PMID- 8729776 TI - [Romberg's facial hemiatrophy: a case report]. AB - The authors report the case of a 45-years-old female patient, with progressive hemiatrophy of the left face and tongue. The laboratorial and image studies were normal, including routine blood exams, cerebral-spinal fluid, hepatic function tests, renal and rheumatic; besides X ray, computerized tomography and magnetic resonance of skull. The electroneuromyography showed peripheral neurogenic alterations of motor trigeminal and hypoglossal nerves, markedly on the left side. Those findings are compatible with Romberg's facial hemiatrophy diagnosis, motivating discussion of several aspects of this rare disease according to the account and literature. PMID- 8729777 TI - Recurrent meningitis in a case of congenital anterior sacral meningocele and agenesis of sacral and coccygeal vertebrae. AB - A rare case of recurrent meningitis due to congenital anterior sacral meningocele and agenesis of the sacral and coccygeal vertebrae is described. An autosomal dominant inheritance is demonstrated for lower cord malformation, and environmental factors (chromic acid or fumes) are discussed. PMID- 8729778 TI - [Granulomatous form of cerebral cryptococcosis: a case report]. AB - Cryptococcal granuloma or toruloma of the central nervous system is very rare, meningoencefalitis or meningitis being most common. The authors present the case of an immunocompetent male patient of 56 years old with initial clinical symptoms of intracranial hypertension due to an occipito-parietal crytococcal granuloma that was removed surgically. The patients was treated post operatively with anphotericin-B and 5-fluorcytosine. Pathophysiology, neuroimaging, diagnosis and treatment of this rare entity are discussed. PMID- 8729779 TI - [Spontaneous tegmental-mesencephalic hematoma: neurobehavioral aspects of the rostral third of the human brainstem]. AB - The tegmentum of the midbrain is a complex area traversed by a number of anatomical and functional systems, including local circuits, ascending activating systems and descending fibers from the cerebral hemispheres. In the present paper we report on the case of a 67-year-old man who suffered a spontaneous central tegmental hemorrhage and was initially supposed to be in coma due to bilateral ptosis and lack of speech and initiative. By the second hospital week, however, he was shown to be able to respond to verbal commands, sit, stand and walk. He died in sepsis one month later. This case shows that the clinical diagnosis of coma may be misleading in certain patients in whom wakefulness is preserved, though concealed from a casual bedside examination due to abulia and ophthalmoplegia. PMID- 8729780 TI - [Coma and transtentorial herniation syndrome due to acute non-expansive hemispheric lesion]. AB - An 81-year-old woman developed left hemiplegia and coma due to a right hemisphere infarct and died 6 days later. When first seen in coma she had the classical signs of descending central herniation in the diencephalic stage. The CT scan of the third day showed a large hypodense area occupying the superficial and deep territories of the middle and anterior cerebral arteries on the left as well as a frontal cortico-subcortical hypodense area indicative of an older infarct on the opposite hemisphere. No mass effects were apparent. She followed a downhill course with signs of brainstem deterioration. A second CT scan a few hours before death revealed the expected pattern of brain shift and herniations. This case adds to the available evidence showing that the clinical signs of encroachment of supratentorial structures upon the basal forebrain can be reproduced by an acute mono-hemispheral lesion without visible mass effects. It indicates, further, that the role of intracranial displacements in the genesis and short-term prognosis of coma remains an unsettled matter. In at least a few number of cases, diaschisis might play a major role. PMID- 8729781 TI - Hemispheric strokes: clinical or surgical management? Intracranial pressure monitoring of a case and review of the literature. AB - The authors report a case of a right hemispheric ischemic infarction on a 77-year old patient that was monitored with an extradural intracranial pressure monitor (Ladd device). We show the huge intracranial hypertension that the brain was submitted despite of the medical treatment. The treatment of hemispheric strokes can be very challenging. The usual antiedema drugs very often prove to be of no help against the mass effect and the high intracranial pressure that some patients present. More recently reports on the literature are showing good results following a decompressive surgery on such ischemic infarcts. PMID- 8729782 TI - [Brainstem hemorrhage after surgical removal of arachnoid cyst of the Sylvian fissure: a case report]. AB - The authors report a case of a hemorrhage of the brainstem after craniotomy for resection of a huge arachnoid cyst of the sylvian fissure on the left hemisphere. The previous simptomatology included clinical signs of increased intracranial pressure, and the computerized tomography showed midline shift. Some factors may contribute to brain hemorrhage post-operatively: cerebral edema, ipsilateral changes in the venous reflux and blood perfusion, although the physiopathology remains obscure. A more careful approach is suggested in such cases with intracranial hypertension. PMID- 8729783 TI - Horner's syndrome and brachial paresis as a complication of lumbar sympathetic block: a case report. AB - An unusual case of Horner's syndrome secondary to a sympathetic block in a patient with chronic adhesive arachnoiditis (CAA) is described. The patient, a 40 year-old white woman, presented with spastic paraplegia, hyperreflexia, bilateral Babinski sign, superficial and deep sensitive hypoaesthesia at the T4 level, in addition to bladder and rectal dysfunction since she was 32. At age of 38 she complained of excessive daily sweating below the T4 level, mostly at night. A 4mL 0.5% bupivacaine lumbar sympathetic block was performed. Within 15 min a right brachial paresis and an ipsilateral Horner's syndrome were noted. Speculatively, an abnormal cephalic spread of the anaesthesic due to a putative erratic space secondary to the CAA may justify the clinical picture even using a relatively small amount of anaesthesic (4 mL). PMID- 8729784 TI - [Benign chondroblastoma of the occipital bone. A case report]. AB - Benign chondroblastoma is a rare bone tumor of immature cartilage cell derivation. This lesion classically occur at the ends of long bones in young persons. Chondroblastoma arising from the occipital bone is extremely rare. We report the case of a 16 year old girl with a benign condroblastoma in the occipital bone, and discuss the clinical, radiological and treatment aspects of this tumor. PMID- 8729785 TI - [Chondromyxoid fibroma of the thoracic spine: a case report and review of the literature]. AB - A case of chondromyxoid fibroma (CMF) arising from the 5th right costovertebral junction and spreading into spinal canal causing spinal cord compression is presented. A myelotomography revealed a complete block at T5 level. The patient underwent a decompressive laminectomy with removal of an epidural tumor. This specimen was sent for pathological examination and interpreted as a CMF. The patient had a neurological improvement, post operative MRI revealed a spinal cord free of compression, and we decided on the follow up of the case. Two years later there was recurrence of the tumor. A posterolateral access by costotransversectomy was made and the lesion was resected. The patient had a neurological improvement which persists on the follow up (two years, at present). Clinical, radiologic and histologic findings, surgical management and recurrence are discussed. The pertinent literature is reviewed. PMID- 8729786 TI - Citalopram, depression and pseudo dementia: a neuropsychological case study. AB - The author presents a case of (depressive) pseudo dementia, commenting on the clinical and neuropsychological findings before and after the use of citalopram, a serotoninergic anti depressive drug. The case portrays the current criticism about the old dichotomy between non-reversible ("functional") and reversible ("organic") dementia. The 73 year old woman initially diagnosed as pseudo demented showed some mild cognitive deterioration in neuropsychological evaluation after the improvement of her depressive symptoms. Some reasons for the divergent findings on pseudo dementia prognosis in the literature are proposed. PMID- 8729788 TI - [Obsessive-compulsive disorder and the basal ganglia]. AB - Obsessive-compulsive disorder (OCD), characterized by obsessions and compulsions, was described as more frequent in patients with primary lesions of the basal ganglia suggesting that these brain structures may be also altered in OCD. The basal ganglia, that were considered important only for the motor control, are known now as crucial for many other mental functions as processing of cognitive experience. Recent studies using magnetic resonance image have found a tendency for smaller caudate nucleus in patients with OCD. Consistently, studies using functional neuroimaging suggest implication of a neurocircuit that includes the orbitalfrontal cortex, the cingulate cortex, caudate nucleus and thalamus in the pathophysiology of OCD. Among several hypotheses formulated to explain these findings, some authors speculated that a deficit of the caudate nucleus function would lead to inadequate filtering of worry inputs which would trigger the orbitalfrontal cortex to develop adaptative responses: the compulsions. PMID- 8729789 TI - [Lymphomatous meningoencephalitis in a patient with HAM/TSP]. PMID- 8729790 TI - [The extension cutaneous-plantar reflex (Babinski, 1896/1898)]. PMID- 8729791 TI - Population review: (3). Migration to from and within the United Kingdom. AB - Monitoring migration trends and patterns is not a straightforward task given the absence of a compulsory United Kingdom-wide system for recording domestic changes of address. Nevertheless, data extracted from the available sources offer a valuable insight into the diverse and complex phenomenon of migration. This article examines international and internal migration phenomenon of migration in the 1980s and 1990s, and looks at the geographical impact of the two in combination on the population of the United Kingdom. This is the third article in the Population Review series. Earlier articles on: (1) Structure and distribution of the population and (2) Are we healthier? appeared in Population Trends issues no. 81 and no. 82 respectively. PMID- 8729792 TI - Which areas are healthiest? AB - This article compares mortality rates by geography and over time. This is done by analysing, for England and Wales, mortality data from 1981 to 1992 by cause, local authority, and ward, using the new OPCS area classifications, and the urban/rural categorisation of 1991 Census enumeration districts based on land use. The proportion of babies who in 1990-92 were lightweight at birth was also examined. Urban areas (particularly purpose-built inner city estates and deprived industrial areas) tend to be the least healthy. Rural and prosperous areas are the most healthy, and the biggest health gains have been made in these. Based on 1992 mortality rates, out of every 100 boys born in 'Ports and industry' areas, 16 would survive to age 85 whereas 24 would do so in the 'Most prosperous' areas. The corresponding figures for girls were 33 and 43. The 'Most prosperous' areas also had the most similar male and female life expectancies, with the difference narrowing throughout the period 1981-1992. In 1990-92 people in 'Ports and industry' areas had the highest male mortality levels for malignant neoplasms, lung cancer, circulatory diseases, ischaemic heart disease and cerebrovascular disease. People in 'Inner London' had the highest levels for respiratory diseases and injury and poisoning. PMID- 8729793 TI - The proportion of married couples who divorce: past patterns and current prospects. AB - This article provides estimates of the proportions of marriages which ended in divorce for the different groups of couples who have married since the 1950s. Over one quarter of all couples who married in the late 1970s and early 1980s had divorced by the end of 1994. As well as giving the overall proportion of couples married in a given year who subsequently divorced, estimates are provided of the corresponding proportions for different subsets of those couples--according to each partner's marital status before marriage, and age at marriage. For some of the higher risk groups--bachelors and spinsters who married as teenagers and divorced men and women who remarried in their early twenties--the proportions who had divorced by 1994 had reached over 40 per cent, and in some cases just over 50 per cent, amongst those who had married between the mid-1960s and the mid-1970s. Finally, a life table analysis is carried out to estimate the proportion of marriages which would end in divorce were the duration--specific divorce rates to remain unchanged at their 1993/94 levels. On this basis, two in five marriages would ultimately end in divorce; just under one half of couples would celebrate their silver wedding, whilst the average length of marriage would be 26 years. PMID- 8729794 TI - Economic activity and mortality of the 1981 Census cohort in the OPCS Longitudinal Study. AB - This article presents mortality from 1981 to 1989 for men and women in England and Wales of working ages by their economic activity at the 1981 Census. Both men and women in employment had lower overall mortality compared with that of all men, while those seeking work had higher mortality. The social class distribution of the unemployed did not explain all their excess deaths. The overall mortality of both employed and unemployed men relative to all men in the 1980s was lower than that of employed and unemployed men in the 1970s (1971-79) relative to all men in the 1970s. PMID- 8729795 TI - Local government reorganisation in Scotland and Wales. AB - This article describes the new local authority structure that comes into effect in Scotland and Wales on 1 April 1996. The relationship between the new 'unitary' authorities and other administrative areas is discussed briefly and some key demographic data are presented. References are given to other sources of statistical information. PMID- 8729796 TI - Isolation and characterization of the d1 domain of Pseudomonas aeruginosa nitrite reductase. AB - Proteolitic digestion of nitrite reductase from Pseudomonas aeruginosa allows to obtain and purify a domain containing only the d1 heme and constituted by two noncovalently bound peptides. This d1 domain catayzes oxygen consumption, and binds carbon monoxide with a kinetic constant slightly higher than the parental dimeric holoenzyme. The capacity to oxidize the physiological substrate, cytochrome c551, is lost, even when the proteolytic c heme domain is added to this reaction mixture. This finding suggests that the two domains do not have a significant affinity for each other, and are kept together only by being part of the same polypeptide. PMID- 8729797 TI - Reactivity of the N-terminal cysteine residues in active and inactive forms of FNR, and O2-responsive, Fe containing transcriptional regulator of Escherichia coli. AB - FNR, the O2-responsive gene regulator of anaerobic respiratory genes in Escherichia coli, contains an N-terminal cluster of four cysteine residues (Cys16 X3-Cys20-X2-Cys23-X5-Cys29), three of which are thought to be involved in the binding of an iron cofactor. The accessibility of the cysteine residues for iodoacetate is known to increase upon switch from the active (anaerobic) to the inactive (aerobic or metal depleted) state. It was analyzed which residues become accessible under either condition. Up to four modified forms, FNR-I, FNR-II, FNR III, and FNR-IV, containing approximately 1, 2, 3.5, and 5 carboxymethyl groups, were obtained either by reaction in vivo and in vitro under conditions of aerobiosis, anaerobiosis, or iron limitation. By N-terminal sequencing, the carboxymethylated cysteine residues were identified. The amount of label in each of the four cysteine residues increased rather uniformly and gradually from FNR-I to FNR-IV irrespective of the condition of labeling; only Cys16 was preferentially labeled to some extent. It is concluded that the four essential cysteine residues change their accessibility in a similar way in the switch from active to inactive (aerobic or metal depleted) FNR, without specific differences in their reaction or function. Potential modes of Fe-binding by the cysteine residues are discussed. In addition, a different type of interaction of Fe(II) with FNR is described. The interaction occurred also in FNR carboxymethylated at approximately three cysteine residues. PMID- 8729798 TI - Nitrogen-15 NMR chemical shifts in oligopeptides coordinated to cobalt(III). AB - Dipeptide, tripeptide, and tetrapeptide complexes with cobalt(III) ions were studied as model compounds for evaluation of 15N NMR chemical shifts induced in proteins upon binding transition metal ions. Coordination of oligopeptides to cobalt(III) resulted in large negative 15N NMR shifts for amine nitrogens (-76 to -32 ppm) and deprotonated amide nitrogens (-47 to -10). Coordination-induced shifts were affected by the nature of moiety at the trans position; the shifts were always larger with a carboxylato oxygen than with an amine nitrogen in the trans position. Thus, coordination-induced 15N NMR shifts provided direct and specific information on the stereochemistry of peptide coordination. Two new complexes, [Co(Gly-gly-gly-glyH(-3))(NH3)2] and Ba[Co(Gly-L-hisH(-2))(NO2)3], were synthesized and their structure was determined by NMR spectroscopy. PMID- 8729799 TI - Structure of 5-hydroxy-2-hydroxymethyl-1-methylpyrid-4-one and 13C NMR relaxation studies of its gadolinium(III) complex. AB - In order to further elucidate the properties and biological behavior of 5-hydroxy 2-hydroxymethyl-1-methylpyrid-4-one (M1), its X-ray structure has been determined, and the ability of its gadolinium complex to enhance the relaxation of 13C nuclei has been examined. X-ray analysis using Mo K alpha radiation shows that M1 crystallizes in the monoclinic space group C2/c with a complex intermolecular array of hydrogen bonding. No water molecules were present within the unit cell. Gd(M1)2NO3 x 3H2O has been prepared and found to be very soluble in water. The effect of low concentrations of Gd(III) on enhancing the 13C relaxation times of M1 was examined. Trace amounts of Gd(NO3)3 x 6H2O resulted in significant decreases in the relaxation time of certain carbon atoms relative to the control measurements, and these data indicate that carbon atoms which bear donor atoms for Gd(III) undergo a significantly greater relaxation than the other carbons. The water solubility and hydrophilic character of this complex suggest that it may prove useful for the determination of metal binding sites on peptides and oligonucleotides. PMID- 8729800 TI - Antitumor activity of the Cu(II)-mitoxantrone complex and its interaction with deoxyribonucleic acid. AB - The effect of the Cu(II)-mitoxantrone complex on the DNA synthesis of HL-60 human leukemia cells has been studied by the technique of isotopic liquid scintillation. The results indicated that the complex shows a stronger ability to inhibit DNA synthesis of the tumor cells, and thus it may become a better antitumor drug. The interaction of mitoxantrone and its Cu(II) complex was studied by the methods of electrochemistry and spectroscopy. The complex gives rise to more changes on the conformation and the double-helical structure of DNA; this is closely related to the antitumor mechanism of the complex. PMID- 8729801 TI - Electron paramagnetic resonance studies of cobalt-substituted angiotensin I converting enzyme. AB - Electron paramagnetic resonance (EPR) spectroscopy has been used to study the metal coordination sphere geometry in the cobalt-substituted Zn-protein angiotensin I-converting enzyme (ACE). It has been shown that ACE contains two distinct metal-binding sites. In the presence of the two structurally different inhibitors, captopril and ramiprilat, it is found that the metal binding sites are nearly structurally identical and are separated more than 10 A from each other. The metal atoms are most likely four- to five-coordinated, and it is argued that the inhibitor binds directly to the metal ion. PMID- 8729802 TI - [Isolated rupture of the tendon of the supra spinatus muscle. Results of 49 surgical repair]. AB - PURPOSE OF THE STUDY: The purpose of this study was to assess the results of surgical repairs in 49 << isolated >> tears of the supra- spinatus in 49 patients, 32 males and 17 females. MATERIAL AND METHODS: The average age at operation was 54,2 years (35 to 72) and the dominant side was injured in 30 patients (12 opposite side and 5 unknown). According to Constant's score the pain was permanent in 23 patients, it occurred at moderate exertion in 21 patients and at important exertion in 5. The average pain duration was 43,4 months (0 to 360 months). The range of motion was normal (40 points) in 24 patients, diminished in 15 (30 points), very diminished (10 points) in 7 and the shoulder was stiff in one patient (2 files were uncomplete). All the patients were disturbed in daily living activity and the strength assessment was disturbed because of the pain. In all cases we performed an anterior acromioplasty. At operation, 39 patients had an isolated tear of the supra-spinatus and 10 an associated tear of the long head of the biceps (7 tendinities, 1 dislocation and 2 ruptures). The size of the rupture was less than 2 cm2 in 21 cases comprised between 2 and 5 cm2 in 27 and greater than 5 cm2 in one case. The surgical procedure was a Neviaser's technique in 4 cases, a single suture in 19, a transosseous suture in 19 and a deltoid muscular flap in 7 cases (large retracted tear), in addition we performed 14 resections of the distal end of the clavicle and 4 tenodesis of the long head of the biceps. RESULTS: The results were assessed according to Constant's score on 46 patients (3 lost to follow-up), the average follow-up was 31,5 months (12 to 86). The average scores were: pain 12,3 points (3 to 15), range of motion 35,5 (14 to 40), daily living activity 17,4 (8 to 20), and strength 11,4 (<> side = 14,8 points). The total Constant's scores were 75,7 per cent (actual score) and 88,3 per cent (weighted score), according to the weighted score 24 patients had 100 per cent; 9 were comprized between 85 and 100, 5 between 65 and 85, and 8 were less than 65 per cent. The average post-operative acromio-humeral height was 9,84 mm (pre-operative height = 11 mm). DISCUSSION - CONCLUSION: If we compare the results in terms of surgical procedure the difference was not statistically significant although the transosseous suture was not so good: single suture 91,5 per cent, deltoid muscular flap 90,9 per cent, Neviaser's technique 87 per cent, and transosseous suture 79,1 per cent. Nevertheless the results are not so good (p = 0,01) if it is an industrial injury and if the post operative acromio-humeral height is diminished. On the other hand, age, sex, duration of the pain, occupation, tenodesis of the long head of the biceps and resection of the distal end of the clavicle don't have any pejorative influence on the results. PMID- 8729803 TI - [Transtrochanteric rotation osteotomies for osteonecrosis of the femoral head. Apropos of 20 cases]. AB - PURPOSE OF THE STUDY: Twenty consecutive rotation osteotomies for idiopatic necrosis of the femoral head were reviewed with an average follow-up of 6,5 years, in order to evaluate an original technique (which uses a nail plate for rotation and fixation of the fragments), and to determine the middle term results (and therefore indications) of anterior and posterior rotation osteotomies. MATERIALS AND METHODS: TECHNIQUE: rotations were achieved by rotating the femoral head with the nail of the nail plate, and without dissection of the posterior vascular bundle. We performed 16 anterior rotation osteotomies (according to Sugioka, with an average rotation of 52 degrees) and 4 posterior rotation osteotomies (described by Kempf, with an average rotation of 77 degrees). Only two patients were lost after 2 years follow-up (with good result), and the radio-clinical outcome of 18 operations at 5 years was known. RESULTS: Global results were : 7 failures, 3 fair and 10 very good or good. In the 4 posterior rotations (Kempf) we achieved 4 very good results, even in Ficats stage 3. In the 16 anterior rotations we could in all cases obtain, on the hip in extension, an almost complete discharge of the necrotic zone, as after osteotomy it was no more in front of the acetabular major bearing zone (defined as an angle of 40 degrees around the apex of the femoral head on the lateral Lequesne view). We obtained 6 good and very good results, 3 fair, and 7 failures requiring a THR. There were 2 factors of poor prognosis : Stage of the necrosis, as we observed 4 failures in the 4 Ficat's stage 3, and only 3 failures in the 12 stage 2. Depth of the necrosis, as we achieved 6 very good and good results and 1 poor in the 7 cases when it was no more than 1/3 of the head diameter. But in the 9 cases where depth was over one third there were 3 fair and 6 poor results. DISCUSSION: Our technique proved to be reliable as it achieved the rotation planned before operation (only one hypo-correction of 15 degrees) and bone fusion, allowing full weight bearing at 3 months in all cases. No extension of the necrotic area was observed. Posterior rotation osteotomy was followed by long term favorable results, may be because it achieves an anatomic discharge of the necrotic zone not only when the hip is in extension, but also when the hip is flexed. Anterior rotation is only recommended when : a rotation not exceeding 60 degrees (therefore without risks for the posterior bundle) allows a discharge of the necrotic zone when the hip is in extension. The necrosis is stage 2. In Stage 3 a progressive arthritis may occur as, in hip flexion, the necrotic sector of the non spherical head comes in front of the acetabular major bearing zone. The depth of the necrosis does not exceed 1/3 of the femoral head, such as in cases of a very large necrosis, mechanical degradation of the non necrotic part of the head may occur, even if discharge of the necrosis is achieved. CONCLUSION: Transtrochanteric rotation osteotomy may delay of a decade or more the occurring of osteoarthritis, if its indications are restricted to patients under 40, suffering from idiopatic necrosis. In our series Sugioka osteotomy gave good results in stage 2 when necrosis depth was no more than 1/3 of the head diameter. Posterior osteotomy allows a better discharge of the necrotic zone and thus may be proposed in less restricted conditions. PMID- 8729804 TI - [Intra and interindividual variations of pelvic mobility in normal adult walk]. AB - PURPOSE OF THE STUDY: Pelvis motion appears as a main human gait component, it is linked to the lower limb joints and to the spine. Current devices, especially the opto-electronical systems, allow quantitative and tri-dimensional gait studies. The purpose of this study was to quantify the pelvic motion individual variability in a sample of healthy subjects. MATERIALS AND METHODS: The study based on a 18 volunters sample. There were 14 men and 4 women, ranged in age from 25 to 37 years. A clinical examination and a AP radiograph of pelvis allowed to include healthy subjects. We used the three-dimensional analysis VICON system with five cameras. Nine records were performed for each subject during a free speed walking. These nine records were distributed on three different days. RESULTS: The step length medianes varied from 1100 to 1600 mm with a significantly (p < 0.05) regression between the step length, the walking speed and the subjects height. Vertical pelvic oscillations varied in this sample from 25 to 60 mm and linked with step length and walking speed. Pelvic rotation around the vertical axis varied from 1.5 to 15 degrees. We did not found regression between this pelvic rotation and the length step. It seems there are three types of pelvic rotation around the vertical axis. At the beginning of the stance phase, in type I, the pelvis is in the transversal plane whereas in the type II, it appears with the maximal rotation. In type III, the value of pelvic rotation is very low. The successive lateral inclinations of pelvic described a complex motion which varied from 1.5 to 9 degrees. The rotation of shoulders around the vertical axis varied from 4 degrees to 13 degrees and the successive inclinations varied from 3.25 degrees to 12 degrees. We did not found any regression between the pelvic and shoulders motion values. DISCUSSION: This study showed that the pelvis motion varied considerably from one subject to another. These variations induce different ways of walking with various consequences on the hip joint and the spine. We suppose that these variations could take a part in etiology of some diseases as hip arthritis or in total hip arthroplasty failure, especially in cup wearing. PMID- 8729805 TI - [Pure dislocations of the tibio-talar joint. Apropos of 9 cases]. AB - PURPOSE OF THE STUDY: The authors relate nine cases of pure traumatic dislocation of the tibio talar joint and propose to evaluate the clinical and radiological results at a long term follow-up (mean 12 years). MATERIAL AND METHODS: The series included 9 patients (7 men and 2 women) the average age was 33,2 years. The injury was a road traffic accident in 5 cases. Pure dislocation variety of the ankle joint was medial and posteromedial in 6 cases. Open skin injury was found in 7 cases. The mean follow-up was 12 years (5-19 years). 6 patients were reviewed by the same surgeon, 6 patients were examined clinically and with ankle X-rays. All patients except one were treated by reduction, immobilization with a plaster cast for 6 to 8 weeks. The joint was examined radiographically to detect the presence of tibio talar diastasis and degenerative arthritis. RESULTS: At term we had two very good results and 3 good results (no pain or pain occasionally). We have found in 5 cases a degenerative arthritis to the ankle joint (joint narrowing <50 per cent in 3 cases, > to 50 per cent in 2 cases). No joint instability was noted at revision. DISCUSSION: Pure traumatic dislocation of the tibio talar joint is a rare injury. Medial and posteromedial variety are not frequent. Immediate gravity is dominated by vascular and septic complications and long term result by degenerative arthritis. CONCLUSION: The authors think that closed dislocations need orthopedic treatment (closed reduction and immobilization with a plaster cast for 6 weeks), on the other hand, open dislocation need surgical treatment (reduction, ligamentous reconstruction and immobilization in a plaster cast for 6 weeks). PMID- 8729806 TI - [Treatment of hip dislocations and subluxations by Petit's splints]. AB - PURPOSE OF THE STUDY: The management of DDH in infants less than one year old is not yet definite. Many ambulant therapeutic methods are described and Pavlik's harness is widely employed. The use of Petit's splints is less well known and we report our experience with 169 cases. MATERIAL AND METHOD: Between 1973 and 1991, Petit's splints were used in 112 cases, following abduction cushions in 55 cases and after Pavlik's harness in 2 cases. 103 hips were dislocated but reducible. This permitted progressive abduction of the hips in order to obtain the reduction of the dislocation and to ensure stabilization by retraction of the slack articular capsule. RESULTS: Only 8 dislocations (4,7 per cent) couldn't be reduced by this procedure and necessitated another treatment. Hips were normal after 5 years of follow up in 118 cases and there were 43 residual dysplasias. 6 post reductional avascular necrosis (3,7 per cent) were recorded as a complication of this method among which only one was a serlous from. DISCUSSION: This ambulatory method for treatment of congenital hip dislocation seems to be forsaken by many authors. Nevertheless its effectiveness has been demonstrated with a complication rate lower than with Pavlik's harness procedure. PMID- 8729807 TI - [Comparison of evaluation methods of the results of congenital clubfoot treatment]. AB - INTRODUCTION: An ideal method for the assessment of the results of treatment in congenital clubfoot should take into account the morphology of the foot and its function, the personal satisfaction of the patient of his parents, interexaminer variability, and be applied for any type of treatment. MATERIAL AND METHODS: We evaluated at- end of growth the results of treatment of 35 patients with unilateral clubfoot, and analyzed them according to 13 different rating scores already published in the literature. The material was divided in 3 groups; 15 feet having undergone multiple surgeries, 10 feet having had only one posteromedial release procedure, and 10 feet having been treated nonoperatively. RESULTS: None of the 13 reviewed rating methods seems to be ideal. The results of treatment, for one taken foot, were often different from one method to the other. DISCUSSION: The high number of rating scores published in literature, is the direct evidence that there is no concensus in the evaluation of the results of treatment in congenital clubfoot. A unanimous new method should be created and should fill all the prerequisites of the ideal method, described above. Some criteria are very important to consider: A iatrogenic deformity or disability, i.e. severe lateral translation of the foot, acquired convex foot, or calcaneus foot, should be severely penalized; hopping on one foot should be the test used to evaluate the strength of triceps surae muscle in children above 5 years of age ; Radiological evidence of talonavicular dislocation is a very important long term prognostic element. CONCLUSION: A new unanimous method for the assessment of the results of clubfoot treatment is necessary. It should be simple, easy to teach, and easy to use. PMID- 8729808 TI - [Femoral head necrosis after intramedullary nailing of the femur in adults]. AB - An avascular necrosis of the femoral head occurred after intra-medullary nailing of a femoral diaphyseal fracture. This complication may be related to the posterior situation of the proximal insertion hole just posterior to the trochanteric notch and piriformis fossa as demonstrated by the CT scan. This situation of the nail may cause injury to posterior branch of the circonflex artery at the time of rod insertion. Even if it appears exceptional in adult patients with intra-medullary nailing, this potential complication and the anatomy of the superior extremity of the femur should be known. PMID- 8729810 TI - [Fracture dislocation of the ankle with retro-tibial luxation of the fibula (Bosworth fracture) . Apropos of a case]. AB - The authors present a case of fracture dislocation of the ankle joint with the proximal part of the fibula entrapped behind the tibia ; Bosworth's fracture. This fracture was described for the first time in 1848 by Hugier, and classified by Bosworth in 1947. In our case, diagnosis was not done in a first time, and closed reduction failed. This fracture was operated on and open reduction was performed. At the present time, the functional result is good. PMID- 8729809 TI - [Cervical spondylolysis in children. Apropos of 4 cases. Review of the literature]. AB - PURPOSE OF THE STUDY: A clinical and radiological study of four cases of spondylolysis in children. Literature review shows the congenital origin of the disorder and how it is to be treated. MATERIAL AND METHODS: We report on four children with cervical spondylolysis. One of the children presented a two-level spondylolysis C5C6. One case was totally asymptomatic and was hazardly discovered. Standard radiographs were used for diagnosis. Conservative treatment was administered in all cases and all patients were immobilized for a few days. DISCUSSION: It has been clearly established that cervical spondylolysis is a congenital disorder often associated with other anomalies: spina bifida, dysplasia of the posterior articular processes. In the child, cervical spondylolysis is only discovered in fifty per cent of Post traumatic cases. The most frequent finding is at C6. A diagnosis can be made based on the radiographs and the CT scan. MRI will only be indicated in spondylolisthesis in order to analyze the disc statement and the degree of canal narrowing. When possible conservative treatment should be adopted. Arthrodesis is reserved for unstable and painful lesions. CONCLUSION: Cervical spondylolysis in the child is a rare congenital disorder. It must be distinguished from the congenital absence of a pedicle and from a pedicle or isthmic fracture. It is a lesion which is normally discovered once the patient has turned adult and is very well long term tolerated. PMID- 8729811 TI - [Fractures of the tibial tuberosity associated with avulsion of the patellar ligament in adolescents]. AB - PURPOSE OF THE STUDY: Avulsion fracture of the tibial tuberosity is considered as a relatively uncommon adolescent injury. The avulsion of the patellar ligament may occur as part of the injury. Two cases of sixteen year old boys with this kind of injury have been reported. Fractures are described using Ogden et al. classification : type I is an avulsion fracture of the most distal portion of the ossification center of the tubercle ; type II, an upward angulation of the lip fragment ; type III, a fracture of the lip fragment with propagation of the fracture-line into the knee joint. Each type is subdivised into A (no comminution) and B (comminution). Frankl et al. in 1990 address the possibility that patellar ligament avulsion may occur as part of the injury (type C). CASE REPORTS: Two sixteen-year old boys suffered from sharp pain in the knee during a jump while playing basketball. Radiographic evaluation demonstrated an avulsion fracture of the tibial tubercle type III C. The osseous fragment was sutured with trans-osseous suture. The fixation was protected with a tension band passed above the patella and distally in the tibia. It was removed at six weeks. One patient was not reviewed and the other had no complaint one year after. DISCUSSION: Avulsion fracture of the tibial tuberosity Ogden type I and II is generally treated by plaster cast. When the patient is unable to extend his knee actively, an avulsion of the patellar ligament should be suspected. In this case, Frankl et al. proposed radiographic evaluation: the distance between the distal end of the patella and the avulsed fragment was noted to increase during flexion. Avulsion of the distal tuberosity usually occurs just prior to complete closure of the proximal tibial physis. Interfragmental transphyseal screws can thus be used safely. In the younger patient or in extensive comminution, tension band wire should be used. The use of a tension band between patella and tibia to protect the ligament reinsertion provise early knee mobilisation (between 0 and 60 degrees). It should be removed within six weeks. CONCLUSION: Avulsion fracture of the tibial tuberosity is rare. When conservative treatment is indicated, it is important to search an avulsion of the patellar ligament. When such an injury is suspected, lateral radiographs of the knee should be made in flexion as well as in extension. We believe that treatment should include open reduction and internal fixation associated with a protect tension band allowing early rehabilitation. PMID- 8729812 TI - [Gluteal posterior approach for surgery of the acetabulum]. AB - PURPOSE OF THE STUDY: This approach was developed to avoid gluteus maximus muscle injury which is divided when using the common Langenbeck-Kocher approach. The posterior perigluteal approach also provides better visualization of the posterior aspect of the acetabulum. MATERIAL AND METHODS: A semi-circular incision is made from the fascia lata at the anterior aspect of the greater trochanter and then directed to the posterio-superior iliac spine. The gluteus maxirnus is then simply retracted medially with blunt retractors. Sixty eight acetabular fractures have been operated with this approach including both mixed and complex fractures of the posterior column. Occasionally fractures of both columns have been operated in two procedures, one posterior and then a second anterior approach. RESULTS: The posterior perigluteal approach preserves gluteus maximus muscle tone. It seems to avoid post-operative ossification. CONCLUSION: We propose an approach which, compare to the Langenbeck- Kocher approach, is less traumatic, and as easy to perform. PMID- 8729814 TI - [Discussion of the Lazarov method, 10 years later]. PMID- 8729813 TI - [Apropos of "Necrotizing fasciitis of the foot"]. PMID- 8729816 TI - ["Francophony"]. PMID- 8729817 TI - [What is the future of dermatologic publications in French?]. PMID- 8729818 TI - [Survival prognosis in pemphigoid. A cohort analysis of 78 patients]. AB - INTRODUCTION: Bullous pemphigoid (BP) is the most frequent autoimmune bullous skin disease and usually affects elderly patients. Despite some conflicting data, vital prognosis is poor in BP, of which mortality rate after one year of treatment varies between 10 and 40 p. 100. To date, the factors influencing this prognosis remain to be determined. PATIENTS AND METHODS: A cohort of 78 consecutive patients with BP (mean age: 80 +- 4 years) has been prospectively selected on a six-year period (1987-1992) in Limoges. The diagnosis of BP was made on clinical criteria (using a standardized questionnaire), direct immunofluorescence (IF) findings (linear deposits of IgG and/or C3 along the basement membrane zone) and confirmed by direct immunoelectron microscopy and/or Western immunoblotting on epidermal extracts. RESULTS: The follow-up analysis (mean duration of follow-up: year of treatment (22 p. 100 in the first three months) with an actuarial survival curve inclined to be horizontal by the end of this first year of treatment. Therefore, we have studied the clinical or immunologic factors susceptible to influence the vital prognosis of BP according to this main criterion: death (or survival) at the end of the first year of treatment. None of the following factors has been found to be significantly linked to the vital prognosis in BP: age, sex, extent of skin lesions at presentation, presence of mucosal lesions, blood eosinophilia, presence of circulating basement membrane zone autoantibodies by indirect IF. The clinical factors of bad prognosis were an altered general condition and a history of coronaropathy. The presence of circulating autoantibodies against BP180 antigen (but not the one of autoantibodies against BP230) as detected by immunoblotting on epidermal extracts was found to be significantly more frequent (50 vs 22 percent) in BP patients who died within the first year of treatment (p < 0.02). DISCUSSION: Although modestly discriminating, the presence of circulating autoantibodies against BP180 represents the first vital prognosis factor demonstrated in BP. This result confirms the growing pathophysiologic importance of the anti-BP180 autoantibodies of which the pathogenic role has been recently formally demonstrated in BP using an animal model. PMID- 8729819 TI - [Quadri-chromic staining in routine cutaneous histopathology (hematoxylin, eosin, saffron and astra blue)]. AB - INTRODUCTION: The routine histologic stains used in dermatopathology are usually hematoxylin-eosin-safran. In order to stain the mucins, specific dyes are necessary. The purpose of this study was to perform a routine quadrichromic stain which included Astra blue as specific dye of mucins. The applications of this hematoxylin-eosin-safran-Astra blue stain were evaluated after 5 years of systematic use. METHODS: The quadrichromic stain was realized by Varistain (Shandon). The time of coloration was 15 minutes for Astra blue pH 2.5, 10 minutes for hematoxylin, 45 seconds for eosin and 4 minutes for safran. A total of 56,000 samples were analyzed using the quadrichromic stain. RESULTS: When mucin was present, it was colored in blue and the contrast with the other dyes was enhanced. Astra blue stained specifically the mucin deposits and it did not modify the other colours. The mucin deposits present in primary and secondary cutaneous mucinoses were most often visible after quadrichromic stain. Because of the variability of the mucin composition, the Astra blue did not react with all mucins. This stain was particularly useful in extramammary Paget's disease and in atrophic malignant papulosis. CONCLUSION: This new stain was easy to perform and allowed a specific coloration of mucin deposits in many cases. The use of four colours therefore improves the results of a routine stain, with a high quality of coloration. PMID- 8729820 TI - [Two-stage excision of extensive cutaneous carcinoma of the face. A retrospective study of 80 patients]. AB - INTRODUCTION: Delayed reconstruction (two-step surgery) comprised of tumor resection and repair three or four days later, after paraffin tissue fixation and histopathological examination, is one of the techniques used for complete excision of spreading cutaneous carcinomas. PATIENTS AND METHODS: In order to evaluate the efficacy and long term prognosis of delayed reconstruction surgery, we carried out a retrospective study of 80 patients treated by this method and (Kaplan- Meier) for correlation with the local recurrence rate: sex, age, anatomic site, histologic type and existence of previous therapy. Long term results have been estimated. Among the 80 patients studied, 76 patients have been followed. The sex-ratio was 0.48. Fifty-two percent of the population was more than seventy years old. Fifty-five percent of the tumors were situated on the nose. In 32,5 p. 100 of the cases, a prior therapy (surgery, electrodessication or radiation therapy) has been carried out. Histological analysis showed 87 p. 100 basal cell carcinomas (of which 26 p. 100 of morpheaform basal cell carcinomas) and 13 p. 100 squamous cell carcinomas. RESULTS: After the first operative step, there was total excision in 69 p. 100 of thecases. In the remaining cases, a second excision was performed with a margin of normal tissue. Recurrences were observed (some precocious recurrences and some late recurrences) in 18.4 p. 100 of the cases. No risk factor correlated significantly with the recurrence rate. However, analysis of the survival graph suggests that the squamous cell carcinomas recurred more often than the basal cell carcinomas. CONCLUSIONS: Although extemporaneous examinations (Mohs' technique) remains the gold standard technique in the management of spreading cutaneous carcinomas of the face, the delayed reconstruction (two-step surgery) is a credible alternative when compared to the recurrence rates reported in the literature with the other techniques (electrodessication, radiation therapy or cryosurgery). PMID- 8729821 TI - [Cutaneous periarteritis nodosa resistant to combined corticosteroids and immunosuppressive agents. Efficacy of treatment with intravenous immunoglobulins]. AB - INTRODUCTION: Cutaneous polyarteritis nodosa is a subset of polyarteritis nodosa (PAN) characterized by a good prognosis because of the absence of severe visceral involvement. Dapsone, colchicin, non steroidal anti-inflammatory drugs are the first line treatments of cutaneous PAN. Corticosteroids are an alternative therapy. CASE-REPORT: A case of cutaneous PAN followed for 6 years is reported. Dapsone, colchicin, corticosteroid and cyclophosphamide have failed to control cutaneous lesions. Intravenous immunoglobulins 400 mg/kg/d five day monthly for 5 months was successful, but a cutaneous relapse occurred 3 months later which was another time controlled with intravenous immunoglobulins. DISCUSSION: Some cases of necrotizing vasculitis have been treated with intravenous immunoglobulins. This is the first reported case of cutaneous PAN treated with intravenous immunoglobulins. Because of the lack of controlled studies, the indications of intravenous immunoglobulins in vasculitis are restricted to those resistant to corticosteroids and immunosuppressive therapy. PMID- 8729822 TI - [Microsporum canis tinea ciliaris and blepharitis]. AB - INTRODUCTION: The involvement of eyelashes and eyelids by dermatophytes is unfrequent. CASE REPORT: We describe such a case in a 48 year old woman, who presented with unilateral blepharitis, resistant to topical treatments with antiseptics, antibiotics and corticosteroids. Diagnosis was suspected by magnifying lens and Wood's light examination; Microsporum canis was isolated from broken eyelashes and scales of annular lesions of eyelids. CONCLUSION: This case emphasizes the interest of mycological examination of eyelashes and eyelids in front of a persisting unilateral blepharitis. PMID- 8729823 TI - [Cutaneous and lymphatic cryptococcosis after transient lymphopenia in an immunocompetent patient]. AB - INTRODUCTION: Cryptococcosis is an opportunistic infection, that rarely occurs in immunocompetent patients. CASE REPORT: We report a case of a 82 year-old woman who suffered from a bacterial pneumonia associated with lymphopenia. During the hospitalisation, she developed a cutaneous and ganglionnary cryptococcosis. No other cause of immunosuppression was found. The lymphopenia disappeared with the healing of the pneumonia treated with systemic antibiotics. Treatment with fluconazole was effective in 2 months on the cryptococcosis. COMMENTS: To our knowledge, this is the first case of cryptococcosis after a transitory lymphopenia, occurring in a immunocompetent patient. PMID- 8729824 TI - [Familial erythermalgia treated with pizotifen]. AB - INTRODUCTION: Primary erythermalgia is a rare acrosyndrome due to paroxystic arteriolar dilatation of extremities that become red and congested with warmth and burning sensation or pain. OBSERVATION: We report herein the observation of a primary erythermalgia affecting 3 degrees in the same family (grandfather, uncle/mother, and children with early onset at 6 years): pizotifene, an antiserotoninergic drug proved to be effective in mother and 14-year-old child who had never drawn benefit from any previous treatment. DISCUSSION: Several etiologies may be responsible for erythermalgia according to the clinical type: primary erythermalgia with pediatric onset, and secondary erythermalgia as a reaction to myeloproliferation, other diseases, drug or heavy metal poisoning. Pathophysiology of primary erythermalgia is not clearly understood and the main concern is therapy: in this respect, pizotifene deserves to be considered as a valuable possibility. PMID- 8729825 TI - [Cutaneous necrosis of the extremities during carbon monoxide poisoning]. AB - INTRODUCTION: Skin lesions may occur in approximately 30 p. 100 of cases of carbon monoxide poisoning. Flushes, erythematous to violaceous blisters on pressure zones due to sweat gland necrosis can be observed in such case. CASE REPORT: We report a case of acral (fingers, ears and nose) superficial cutaneous necrosis due to carbon monoxide poisoning. DISCUSSION: Such a complication had never been reported as a cutaneous manifestation of acute carbon monoxide intoxication. Also rare, acral necrotic lesions in a comatose or obnubilated patient should suggest this diagnosis. PMID- 8729826 TI - [Tumoral sarcoidosis]. AB - INTRODUCTION: We report here an unusual tumoral form of cutaneous sarcoidosis which lead to diagnostical difficulties. CASE REPORT: A 56 year-old woman was hospitalized for a large lumbo-sacral tumor which size had rapidly increased. The tumor was purplish-red, firm, measured 10 x 20 cm and the patient was unable to sit normally. The biopsies demonstrated a dermohypodermal epithelioid and giant cell granuloma without necrosis. We ruled out an infectious granuloma and a foreign body granuloma. There were no extra-cutaneous localizations. This sarcoidal tumor cured with systemic corticosteroids. DISCUSSION: Tumoral cutaneous forms of sarcoidosis are uncommon; 3 other cases of the literature are reported. Before to state positively this forms it is obligatory to rule out all the other aetiologies of sarcoidal granuloma. The existence of extra-cutaneous localizations and multiple skin localizations is helpful to diagnose this uncommon type of sarcoidosis. PMID- 8729827 TI - [Necrolytic migrating erythema without glucagonoma]. AB - INTRODUCTION: The glucagonoma syndrome is an uncommon but well-known entity associating erythema necroticans migrans (ENM) with glucagonoma. CASE REPORT: A 43-year-old man with a past history of alcoholic cirrhosis and ascitis was hospitalized for skin disorders which had developed over the past 4 months. Centrifugal erythematous skin lesions were observed, some with non-turgid bullae and marginal desquamation, others with an erosive center. Lesions first appeared on the hands then diffused widely without involving the periorifical areas, folds and lower limbs. Laboratory results revealed an anemia, hypovitaminosis K, cholestatic liver failure, a beta-gamma block and low zinc levels. Histology study of the skin biopsy demonstrated a << Neapolitan trench >> image suggestive of ENM. A paraneoplasic syndrome and pancreas tumor were not found. Despite supplementation with zinc, amino acids and vitamins, the patient died from his liver disease. DISCUSSION: There have been 4 cases of ENM reported in the literature, all in cirrhosis patients, two of which had low zinc levels. Our case is thus the third with cirrhosis and low zinc. Bazex's syndrome, acrodermatitis enteropathica, annular chronic lupus erythematosis and annular superficial pemphigus were eliminated as possible diagnoses. The failure of zinc and amino acid supplementation would favor the secondary nature of the zinc deficiency and the predominant role of cirrhosis in this skin disease. The possible role of essential fatty acids in ENM is raised. PMID- 8729828 TI - [Calcified intertrigo, a rare cause of cutaneous calcinosis]. AB - INTRODUCTION: Calcinosis cutis is classically described in patients presenting chronic renal failure with secondary hyperparathyroidism. There are three clinical types described in the literature: cutaneous necrosis (secondary to vascular calcification), panniculitis with calcification of the adipose tissue and necrosis of the skin and dermo-hypodermic calcinosis without necrosis usually called metastatic calcinosis. The latter can affect all tissues but skin involvement is rare. CASE REPORT: A case of calcinosis cutis is presented in a 50 years old woman with moderate secondary hyperparathyroidism and chronic renal failure caused by renal amyloidosis. She was referred by the renal unit to our service for erythematous, woody-hard, infiltrated plaques with petechial purpura satellite lesions, involving the flexural areas, that appeared within a few weeks. DISCUSSION: This observation is original by the topography of the cutaneous lesions the involvement of the fexural areas is unusual and by the histologic aspect of pseudoxanthome elastic-like. We must notice that the hyperparathyroidism had always been very moderate (normal parathormonemia) compared to the rapid development of an important dermic calcinosis. PMID- 8729829 TI - [Bullous manifestations of Kaposi disease associated with AIDS]. AB - INTRODUCTION: We report the apparently first description of bullous Kaposi's sarcoma in a patient with AIDS. CASE REPORT: Kaposi's sarcoma was observed in a HIV positive homosexual who had developed AIDS. The delay between the development of Kaposi's sarcoma and the appearance of bullae was 6 months. Bullae occurred after oedema and necrosis of the subepidermic derma. DISCUSSION: This unusual clinical presentation of Kaposi's syndrome in an AIDS patient led to rapid tumor extention and would thus suggest a rapidly unfavourable prognosis. PMID- 8729830 TI - [A case for diagnosis: pilomatrixoma]. PMID- 8729831 TI - [A case for diagnosis: erosive pustular dermatosis of the scalp]. PMID- 8729832 TI - [Transient acantholytic dermatosis (Grover disease)]. PMID- 8729833 TI - [Human immunodeficiency type 1 virus (HIV-1) primary infection]. PMID- 8729834 TI - [Monthly question: administration at home of topical treatment by private nurses]. PMID- 8729835 TI - [Positive corneal findings of Propioni bacterium acnes and keratitis]. AB - PURPOSE: Propioni bacterium Acnes may be isolated from corneal specimens. Its presence usually evokes a neighbouring tissue contamination. Nevertheless we have tried to find arguments in favour of a possible pathological responsibility of this microorganism in corneal affections. MATERIAL AND METHODS: A retrospective study was conducted in thirty patients with proved P. acnes corneal infection. The specimens were collected immediately during the first examination in the emergency ward. The research of anaerobic bacteria was made for each corneal specimen. RESULTS: We studied nineteen corneal abscesses and eleven corneal ulcers. Ninety-one percent of the cases presented an associated irritating factor. In decreasing order we found: wearing of soft contact lenses (36%), foreign corneal body still in situ or recently removed (21%), recurrent corneal erosions (9%), palpebral surgery (6%), herpetic keratitis (6%) or rosacea (3%). P. acnes was the only microorganism identified in 97% of the cases. Starting treatment before bacteriological results were known to be effective in each case. CONCLUSIONS: These elements are an argument for the pathological role of P. acnes in corneal infections when combined with favourable local conditions. Its presence does not always mean actual contamination. PMID- 8729836 TI - [Retinal vasculitis]. AB - PURPOSE: The purpose of this study is to determine aetiologies of vasculitis of retina in Morocco. MATERIAL AND METHODS: This is a retrospective study on 108 cases of vasculitis of retina admitted in the ophthalmology department, for a 5 year period (1987-1993). Our patients had a complete ophthalmological examination : fluorescine angiography, colour vision, general examination and biological study. RESULTS: Many aetiologies have been found, predominantly Behcet's disease. The neovascularization of retina which complicates the vasculitis was detected in 30 % of our patients. A laser photocoagulation was performed on these cases, sometimes a vitrectomy was necessary when a haemorrhage of vitreous body occurred, complicating the neo-vessels of retina. The therapeutics received by our patients was based on the corticotherapy. An immunosuppression treatment was prescribed in most cases of Behcet's disease and in severe vasculitis. Outcome after treatment has been satisfactory, but many patients relapse very frequently, mainly those with Behcet's disease. CONCLUSION: Causes of vascular retinitis are variable, most of them are due to Behcet's disease. PMID- 8729837 TI - [Treatment with silicone oil in complicated retinal detachment. Anatomopathological test of 2 enucleated eyes]. AB - Two eyes with a complicated retinal detachment were operated using intraocular silicone oil injection. They were enucleated 33 and 12 months after the operation because of painful secondary glaucoma and corneal dystrophy. Using light and electron microscopy, silicone oil vacuoles were observed in the corneal depth, the iris stroma, the trabecular meshwork and in lens remnants. There was an angle closure due to iridocorneal synechia. The same vacuoles were recognized in epiretinal membranes, in the internal and external layers of the retina and in the pigment epithelium. When the retina was detached, silicone oil vacuoles were present in the choroidal vessels. PMID- 8729838 TI - [Color Doppler ultrasonography in the study of orbital and ocular vascular diseases]. AB - Color Doppler flow imaging was performed in 10 healthy controls and in 32 patients with orbital and retinal vascular diseases in order to investigate ocular arterial supply. Abnormal changes of the orbital blood flow observed in case of dural fistula, orbital varicosis and choroidal melanoma is described. In patients with central retinal vein occlusion, substantial arterial flow disturbances were found, such as the reduction of the retinal artery systolic peak flow (p < 0.55) and absence of diastolic flow (p < 0.001). Abnormal changes of the retinal arterial blood flow (reduction of the systolic peak flow or even extinction of the Doppler signal) were also detected in patients with occlusion of the central retinal artery and in some cases of severe stenosis or occlusion of the internal carotid artery. The color Doppler flow imaging appears as a promising technique for the investigation of the vascular diseases of the eye. It may provide useful clues for the differential diagnosis of optic neuropathies and allows haemodynamic characterization of different vascular diseases of the eye such as diabetic retinopathy and glaucoma. PMID- 8729839 TI - [Phacoemulsification and topical anesthesia. Apropos of 40 cases]. AB - BACKGROUND: To evaluate topical anaesthesia for cataract surgery. METHODS: Thirty nine patients (40 eyes) underwent cataract surgery by phacoemulsification with topical anaesthesia consisting in instillations of tetracaine eyedrops. Thirty four eyes were implanted with a foldable lens and 6 eyes received a PMMA lens. Final visual acuity (1 month minimum follow-up), patients reactions (sensibility, pain), anaesthetic and surgery difficulties were specially studied. RESULTS: Mean initial corrected visual acuity was 0.21. Mean final corrected visual acuity was 0.72; 90 % of the subjects experienced no pain. Implantation was the most sensitive moment. Eye mobility was not a problem except during capsulhorexis which had to be done with forceps. CONCLUSION: Simplicity of topical anaesthesia eliminates the complications of the other anaesthetic methods. It is necessary to have a close patient-surgeon relationship. Carefully chosen topical anaesthesia is an interesting alternative. PMID- 8729840 TI - [Dynamic viscosity and corneal endothelial protection with Healonid, Healon GV, Provisc and Viscoat during phacoemulsification]. AB - PURPOSE: To demonstrate the utility of a viscoelastic substance which maintains high viscosity during phacoemulsification, we conducted a double study. METHODS: We compared in vitro the corneal endothelial protection offered by Healon, Healon GV, Provisc and Viscoat after <> phacoemulsification. Eight samples of ten to fifteen freshly enucleated pig eyes were operated; the first four groups were the controls, whereas the other four tested the different viscous solutions. In parallel, we determined the rheologic properties of the four viscoelastic substances in a physicS laboratory. To measure the percentage of corneal endothelial cell damage, we used the Janus Green spectrometric technique. RESULTS: Corneal endothelial cell damage averaged 31.67% in eyes in which no viscoelastic substance was used. Damage was 14.29% in specimens that received Healon, 12.85% with Healon GV, 2.48% in the Viscoat group and 15.43 in those that received Provisc. There is a significant difference (p <0.001) between the Viscoat group and all other samples. The values of the viscosity at different shear rates and the graph of viscosities as a function of shear rate of the four viscous solutions are given. CONCLUSION: Viscoat has a very high dynamic viscosity at high shear rate. This property and a poor cohesion provide a better corneal endothelial protection during in vitro phacoemulsification. PMID- 8729841 TI - [Paradoxical aggravation of carotid cavernous fistula]. AB - A case of spontaneous thrombosis of carotid cavernous fistula, preceded by paradoxical worsening of ocular symptoms occurred in a 74-year-old patient. Clinical examination, and paraclinical investigation if necessary (echo-Doppler and arteriography) allow to distinguish spontaneous resolution from increased blood flow through the fistula. PMID- 8729842 TI - [Human periorbital dirofilariasis]. AB - We present the case of a 72 year-old-woman with recurrent periocular inflamatory mass caused by an infection with Dirofilaria repens. The zoonotic infection is spreading by mosquito vectors from dogs to humans. Residence in endemic areas (ex USSR, Italy, Sri Lanka, Southeastern United States) should always be suspected in patients with this type of symptomatology. The treatment is curative by the extraction of the pseudotumoral mass. PMID- 8729843 TI - [Dragged disk syndrome. Review of etiologies. Apropos of a case]. AB - A young woman presented with an elevated, retinal yellow-white lesion in mid periphery of one eye, associated with a dragged disc. Different aetiologies were discussed including retinopathy of prematurity, familial exudative vitreoretinopathy, congenital retinal folds, incontinentia pigmenti, Eales' disease, toxocariasis, combined hamartoma of the retina and the pigment epithelium. PMID- 8729844 TI - [Involvement of the connective tissue of the eyelids in pachydermoperiostosis]. AB - A 50-year old man presented with pachydermoperiostosis. Eyelid histopathology is described. Clinical features of this uncommon disorder are reported. PMID- 8729846 TI - [Herpetic uveitis]. PMID- 8729845 TI - [The excito-toxicity theory of glaucomatous neuropathy. General review]. PMID- 8729847 TI - [VLA and alpha6, beta4 integrins. Expression in normal and neoplastic human tissues]. AB - Among the cellular adhesion molecules, the integrin family, more particularly the VLA (Very late antigen) integrins, is currently the subject of numerous investigations in pathology. These integrins are involved in cell-cell contact or cell-matrix adhesions. During neoplastic diseases, cellular expression of integrins changes and a study of the modifications could allow a new etiopathogenic approach carcinogenesis and metastatic phenomena. New prognostic factors may be defined in tumor pathology. We describe the general structure of integrins and the mechanisms of their binding with matricial ligands and with cytoskeleton. The expression of VLA integrins and the alpha6beta4 heterodimer on normal and neoplastic human tissues is then described. Finally, we describe the involvement of these proteins in tumor progression and tissue invasion. PMID- 8729848 TI - [Alveolar sarcomas of soft tissues. Immunohistochemical, ultrastructural study and flow cytometry]. AB - Alveolar soft tissue sarcoma is an unusual tumor, known to have a poor prognosis. Although a muscular origin has been supported by most authors, the histogenesis of such tumors remains unclear. We report a case of alveolar soft tissue sarcoma with histological, ultrastructural, immunohistochemical and flow cytometry study. Pour results support a myogenic origin of these tumors and demonstrate its aneuploid nature. Pulmonary metastasis occurred early in spite of intensive chemotherapy and surgical removal of the tumor. PMID- 8729849 TI - [Study of the reactivity of giant cell tumors of bones to different lectins]. AB - The authors have examined 18 cases of giant cell bone tumours using various lectins. Type I stromal cells resembling fibroblasts and participating in the construction of matrix were Con A positive. The intensity of Con A binding and the number of positive cells increased with increasing stage. The nature and intensity of PNA-lectin binding in giant cells, after digestion with neuraminidase, varied according to the stage. While in Group I giant cells, 30 % of cells were negative and, in the great majority of cells, the reaction was limited to a clearly defined segment of the membrane, in Group III tumours, the proportion of negative cells was approximately 1 % and a diffuse and very intense intracytoplasmic reaction was observed in 60 % of the positive cells. According to the authors, the histochemical study of lectins could help classify giant cell tumours. PMID- 8729850 TI - Primary causes of fetal deaths in Athens a clinicopathological study. AB - The authors studied 239 cases of fetal death, occurring before the 24th week of gestation, during a five-year period, from 1989 to 1993. Based on clinical information and autopsy findings, their cases were classified as follows: 1) amniotic infection syndrome; 2) congenital anomalies; 3) disorders of the placenta and adnexae, and 4) others. The causes of fetal death were determined and evaluated statistically. In 83 cases (39 %), only autopsy revealed the cause of fetal death. The correlation of clinical and pathological findings showed that autopsy constitutes an important source of information for the determination of causes of fetal death, as well as for future genetic counselling. Moreover, it can broaden our knowledge about perinatal mortality, by correctly interpreting clinical and complementary investigation data. PMID- 8729851 TI - [Extracerebral metastases of a glioblastoma, in the absence of surgery]. AB - A 50 y.o. male presented with a right parietal tumor which was a glioblastoma on stereotactic biopsy. He was treated by radiation and steroids, with clinical improvement. Four months later, he presented with a left preauricular mass and cervical lymphadenopathy. CT scan showed destruction of the left mastoid and filling of the left tympanic cavity. One month later, he suffered progressive dyspnea. Chest X ray showed a mediastinal mass on the right side and numerous bilateral interstitial opacities in the lungs. A bronchial biopsy was inconclusive. His general condition worsened, and he died. Postmortem showed continuous neoplastic infiltration of the left part of the base of skull, extending into the neck. Numerous metastases were present in mediastinal lymph nodes, lung parenchyma, pleura and pleural aspect of the diaphragm. There were no subdiaphragmatic metastases. Neuropathological examination confirmed a poorly differentiated highly malignant glioblastoma with severe necrosis involving the internal part of the parietal lobe extending to the dura mater of the convexity and falx cerebri with invasion of the superior longitudinal sinus which was entirely occluded. The biopsy scar was not infiltrated. Visceral tumors were morphologically identical to the brain tumor. They were strongly GFAP positive and cytokeratin negative. Extraneural metastases of glioblastoma in the absence of surgery are uncommon in adults. Involvement of the dura mater and/or superior longitudinal sinus is an almost constant feature. In our case, this may have led to invasion of the base of skull and secondary regional, lymphatic, and hematogenous spread. PMID- 8729852 TI - Squamous cell carcinoma developing in an odontogenic keratocyst. AB - A case of squamous cell carcinoma arising in an odontogenic keratocyst is presented. Although the patient was treated by wide local excision of the tumor, with radical neck dissection, and although the regional lymph nodes were free of metastatic deposits, the carcinoma recurred locally two months later and the patient died two years after the first operation with pulmonary metastases. PMID- 8729853 TI - [Atrophic thyroiditis: an unusual course of a Basedow-like variation of Hashimoto disease. Apropos of a case]. AB - This case concerns a 27-year-old man with a very high level of anti-thyroid antibody and pretibial elephantiasis myxedema which developed progressively over several years following subtotal thyroidectomy for << hashitoxicosis >>. Complementary resection of the thyroid stump was performed ; under microscopic examination this stump presented an aspect of atrophic thyroiditis. This unusual development towards atrophic thyroiditis raises the problem of the relations between the various forms autoimmune thyroiditis. PMID- 8729854 TI - [Transposition of great arteries. The reality of its cure today]. PMID- 8729855 TI - [Efficacy of repeat coronary dilatation in patients with restenosis of the anterior descending artery]. AB - PURPOSE: To analyze the clinical characteristics, acute and long term results of repeat percutaneous transluminal coronary angioplasty (PTCA) for restenosis of the proximal left anterior descending coronary artery (LDA) METHODS: We studied 113 patients, 79% male, mean age 59 years, 49.5% had stable angina, with single vessel proximal LDA disease undergoing repeat PTCA for a first restenosis from January/88 to December/92. We examined the in hospital outcome (success rate and complications) and long term follow up (angina status, occurrence of myocardial infarction (MI), death and need for repeat PTCA or coronary artery bypass graft (CABG) of this subgroup. RESULTS: Primary success was 96%. Complications included: 1% MI, 1% emergency CABG and 1% procedural death. Follow up data (mean 42 months) was available in 102 (94%) out of 109 patients with successful repeat PTCA: 64 (63%) patients were asymptomatic, 5 (5%) had a MI and 28 (27%) required repeat PTCA or CABG. Actuarial 5 year freedom from death was 94%, freedom from death and MI was 91% and freedom from death, MI and repeat PTCA or CABG was 52%. CONCLUSION: Repeat PTCA is an effective treatment for proximal LAD restenosis with a high success rate, low incidence of procedural complications and provides excellent long term cardiac survival, however repeat revascularization is frequently required. PMID- 8729856 TI - [Nutritional status of children with congenital heart disease and left-to-right shunt. The importance of the presence of pulmonary hypertension]. AB - PURPOSE: To assess the nutritional status of children with congenital heart disease with left-to-right shunt and the nutritional disturbances related to the presence of pulmonary hypertension (PH). METHODS: In a prospective study, the nutritional status of 30 children with left-to-right shunt (16 with and 14 without PH) was determined by anthropometric parameters and plasma proteins albumin, transferrin and prealbumin. Frequency and extent of nutritional disturbances were analysed in relation to the presence of PH. A control group of 20 well-nourished children was formed, aiming to compare the values of the serum proteins. RESULTS: The nutritional classification according to Waterlow's criteria showed an overall prevalence of malnutrition of 83.3%, which was higher in those with PH (p = 0.0140). This patient's group was more wasted than the group without PH (median z score of weight for height -1.65 vs -0.78). There was a high occurrence of measurements of arm fat area below the 5th percentile, which were similar in both groups. Serum levels of albumin and prealbumin were significantly lower in children with congenital heart disease than in the control group and transferrin values were similar in both. CONCLUSION: The prevalence of malnutrition was high. The presence of PH was associated with higher nutritional disturbance. The nutritional assessment may be a good way to identify diagnostic groups at particular risk of failure to thrive. This can be useful in planning a management which ensures these patients to achieve adequate nutritional supplementation in early life. PMID- 8729857 TI - [Assisted circulation to the right ventricle through pulmonary counterpulsation using a biological method]. AB - PURPOSE: To study the hemodynamic effects of latissimus dorsi dynamic pulmonaroplasty in open chest animals. METHODS: Six anesthetized mongrel dogs were subjected to diastolic counterpulsation using electrically stimulated latissimus dorsi muscle flap wrapped around the aortic and pulmonary arteries roots and gated to the surface electrocardiogram. Aortic and Pulmonanary pressures as well as cardiac output and cardiac index were measured. RESULTS: Diastolic counterpulsation resulted in a significant increase in cardiac output (from 2.35 +/- 0.26 to 2.45 +/- 0.28 l/min) (p < 0.005) and cardiac index (from 0.108 +/- 0.020 to 0.113 +/- 0.020 l/min/kg) (p < 0.05). The diastolic pulmonary arterial efficiency index showed a significant increase when latissimus dorsi stimulation was on (from 8.37 +/- 0.60 to 11.65 +/- 0.83 mmHg); (p < 0.005). CONCLUSION: Latissimus dorsi dynamic pulmonaroplasty provides an effective means of arterial counter pulsation in open chest dogs. PMID- 8729858 TI - [Magnetic resonance imaging in chronic Chagas cardiopathy. Correlation with endomyocardial biopsy findings]. AB - PURPOSE: To study the correlation between magnetic resonance imaging of the heart and right ventricle endomyocardial biopsy results in chronic Chagas' heart disease. METHODS: Ten patients with Chagas' disease, mean age 47 +/- 7 years, all males, in congestive heart failure with New York Heart Association class II (2 patients), III (6) and IV (2) were studied. Mean left ventricular ejection fraction was at echocardiogram 36 +/- 6%. The patients were submitted to right ventricular endomyocardial biopsy and magnetic resonance imaging of the heart. The results of this group were compared with a control group of patients with idiopathic dilated cardiomyopathy, with mean age of 46 +/- 10 years and left ventricular ejection fraction of 30 +/- 4%, in heart failure with functional class II (1 patient), III (5) and IV (1). RESULTS: All patients with Chagas' heart disease presented an increase in magnetic ressonance imaging signal of the heart after gadolinium use. The septal signal intensity changed from 0.87 +/- 0.06 to 1.54 +/- 0.16 (p < 0.001). In the control group the mean septal signal intensity was 0.93 +/- 0.07 before and 0.89 +/- 0.06 after the gadolinium (p = ns). Eight patients of the Chagas' disease group had biopsy proven myocarditis and two had borderline myocarditis. However, only one patient of the control group had diagnosis of borderline myocarditis. CONCLUSION: Myocarditis is frequently found in Chagas' heart disease patients and who unlike controls present a significant increase in myocardial signal intensity after gadolinium infusion. The magnetic resonance imaging of the heart seems a promising alternative method for the diagnosis of an inflammatory process in Chagas' heart disease. PMID- 8729859 TI - [Influence of afterload on the rate of variation of developed pressure (dT/dt). Study in isolated cardiac muscle]. AB - PURPOSE: To evaluate the effects of afterload on peak rate of tension rise (dT/dt) in the isolated muscle. METHODS: Left ventricular papillary muscles from Wistar rats were studied in isometric and isotonic afterloaded contractions. Muscles were analised in Krebs-Henseleit solution with calcium concentration of 2.52mM at 28 degrees C. The resting muscle length (preload) was maintained constant. The peak isometric developed tension (DT) and dT/dt were measured during increases of afterload (25, 50, 75 and 100% from DT). RESULTS: A rise in afterload corresponding to 50, 75 and 100% of DT, did not cause an increase in dT/dt values (p > 0.05). The dT/dt value decreased (p < 0.05) when afterload was changed from 75% to 25% of DT. CONCLUSION: The data suggest that an increase in the afterload from 50% of the DT did not promote changes in the dT/dt. PMID- 8729860 TI - [Diagnosis using nuclear medicine of anterior descending artery stenosis in patients with left bundle branch block]. AB - PURPOSE: To evaluate septal perfusion and contractility in patients with left bundle branch block (LBBB). METHODS: Twenty patients were submitted to myocardial scintigraphy with Tecnecium-99m isontrile after exercise and dipyridamole infusion. The septal contractility was observed during the radionuclide ventriculography. All patients were submitted to coronariography. RESULTS: Twenty patients were studied and 17 (85%) had a normal left anterior descending (LAD) artery. Nine (53%) patients had abnormalities in septal perfusion after the exercise and 2 (12%) after dipyridamole infusion. All patients had abnormalities in septal contractility during the radionuclide ventriculography. CONCLUSION: The myocardial scintigraphy after dipyridamole infusion showed better specificity in the diagnosis of a LAD lesion. The radionuclide ventriculography was an efficient method to evaluate the septal contraction abnormalities in patients with LBBB. PMID- 8729862 TI - [Role of magnesium in the management of cardiopathy]. PMID- 8729863 TI - [Coronary angioplasty of proximal lesions of the anterior descending artery]. PMID- 8729861 TI - [Severe pulmonary thromboembolism caused by contraceptives in adolescents]. AB - A 16 year-old female patient on oral contraceptive (gynera ethynilestradiol+gestadone) for 4 months, presented with acute and severe pulmonary thromboembolism (PT), and was successfully treated with IV streptokinase. The contraceptive use was an importante risk factor for PT and the thrombolytic therapy provided early clinical improvement. Blood pressure, heart rate, and hypoxemia, along with echocardiography and pulmonary arteriography abnormalities, improved after this treatment. PMID- 8729864 TI - [Current concepts on the treatment of bronchial asthma]. PMID- 8729865 TI - [Consensus SOCESP-SBC on electrocardiography by Holter system]. PMID- 8729867 TI - [Consensus SOCESP-SBC on echocardiography]. PMID- 8729866 TI - [Consensus SOCESP-SBC on magnetic resonance imaging in cardiology]. PMID- 8729868 TI - [Consensus SOCESP-SBC on nuclear medicine]. PMID- 8729869 TI - The Eclampsia trial, a model of international collaborative study with worldwide benefits. PMID- 8729870 TI - Humoral immunity in Hansen's disease. AB - For many years immune response in leprosy has been studied. Since 1960 several reports dealing with humoral immunity have been described in the literature. Different autoantibody rates occur in leprosy. There is an increase in the prevalence of autoantibodies in elderly patients with long standing disease, in lepromatous leprosy and in those with reactional states. The differences in rates among various studies are attributed to different methods and variations among patient samples concerning age, gender, polar forms, therapy and other elements. The prevalence of numerous antibodies, immune complexes, cryoglobulins and complement levels have been studied by many authors. This also highlights the importance of the more recent reviews of anti-Mycobacterium leprae glycolipid antibodies such as the anti-phenolic glycolipid-I antibodies in which titers are variable and depend on genetic factors. PMID- 8729871 TI - Crossed testicular ectopia. AB - Crossed testicular ectopia (CTE) is a rare anomaly, characterized by migration of one testis towards the opposite inguinal canal. Presented here is a case of crossed ectopia of the right testis, treated by extraperitoneal transposition of the gonad and right orchiopexy. Embriology and surgical findings suggest that CTE is a common consequence of many unclear ethiologic factors, specially mechanical ones, and can be associated with Muller duct persistence. Review of literature suggests a classification of CTE into 3 types: I--associated with inguinal hernia alone; II--associated with persistent mullerian remnants; III--associated with other anomalies without mullerian remnants. Treatment includes transeptal orchiopexy or extraperitoneal transposition of the testis, research for mullerian remnants and other anomalies, and long term postoperative follow-up, due to the risk of becoming malignant. PMID- 8729872 TI - Enhancement of liver size by stimulation of intact rat liver with exogenous hepatotrophic factors. AB - In mammals, liver size is related to animal body weight at the 2.5 to 3% proportion, a ratio mediated by the afflux of hepatotrophic factors. Formulas capable of modifying this ratio have been developed in previous studies on the rat, with enhancement of liver size brought about by intraperitoneal (portal) infusion of exogenous factors such as glucose, amino acids, insulin, glucagon, vitamins, electrolytes, and triiodothyronine. However, the efficacy of these formulations was accompanied by increased animal mortality (PARRA et al.). The present study, which was carried out with small methodological modifications on a larger number of rats using daily intraperitoneal injections of a solution of exogenous hepatotrophic factors (40 ml/kg) for seven days, confirms the previous findings, with a 114.16 +/- 7.90% enhancement of liver size beyond the expected value for the body weight of the animal. However, the problem of animal mortality was not fully resolved. PMID- 8729873 TI - Infection by the human papillomavirus in teenagers sexually active: clinic and subclinic manifestations. AB - This research studies the association of the cervical condyloma with the intraepithelial neoplasia, during sex activity, pregnancy, diagnose methods, cytology and colposcopy in teenagers. The objective of this research is to study the propaedeutics for the uterine coli condyloma diagnose in a group of teenagers. For this purpose, we have studied 131 teenagers sexually active with ages between 14 and 19 years and presenting histologically confirmed uterine coli condyloma. Association with intraepithelial neoplasia, sexual activity duration, method of diagnose, pregnancy analysis, cytology and colposcopy results and association with vulva and vagina injuries were evaluated. The high association rate with condyloma and intraepithelial neoplasia after a short time of sexual activity and the none presence of macroscopic warts in the genital organs in 80% of cases presenting cervical condyloma, demonstrate that: a more careful investigation with colposcopy and biopsy of the inferior genitals of the women teenagers sexually active is needed, when presenting modified cervical cytology. PMID- 8729874 TI - Lymphoma of unknown origin located in paravertebral muscles: an unusual cause of low back pain in children. AB - The authors report a case of an adolescent with a poorly differentiated lymphoma of unknown origin located at paravertebral muscles, whose presently symptom was low back pain. PMID- 8729875 TI - Congenital papillomas and papillomatoses associated with the human papilloma virus (HPV)-report on 5 cases. AB - The authors present a study of five cases of vulvar congenital papillomas and papillomatoses in stillborns and neonates dead upon birth. The studied material was collected from five necropsies. The histopathological evaluation showed hyperkeratosis, acanthosis, papillomatosis, perinuclear haloes, and nuclear abnormalities. In three of the cases, the electron microscopy identified nuclear and cytoplasmatic viral particles ranging from 40 to 60 nm in size, compatible with HPV. The immunohistochemical study of those lesions showed nuclear and cytoplasmatic positivity. The authors concluded that the presence of viral particles suggestive of HPV added to the immunopositivity indicated the possibility of viral infection. PMID- 8729876 TI - Osteoporosis 1995. Basic diagnosis and therapeutic elements for a national consensus proposal. Sao Paulo, Brazil, May 12-13, 1995. PMID- 8729877 TI - Prophylactic and therapeutic efficacy of an avian antitoxin in ricin intoxication. AB - This study was designed to demonstrate the efficacy in animals of a ricin antitoxin consisting of purified avian antibodies. Antitoxins consisting of avian antibodies have significant advantages when compared to conventional mammalian (e.g. horse) antibodies; (a) avian antibodies do not fix human complement, eliminating associated inflammatory reactions, and, (b) avian antibodies can be manufactured more economically. Ricin toxoid was injected into laying hens followed by collection of eggs bearing hyperimmune immunoglobulin. Immunoglobulin was extracted from yolks and purified by immunoaffinity chromatography. In a mouse model for toxin neutralization it was shown that immunoaffinity purified ricin antibodies could prevent ricin lethality. Furthermore, it was shown that passive antibody treatment leads to active ricin immunization in animals given lethal ricin doses. Highly purified avian antibodies, as developed in this study, should offer enhanced clinical effectiveness, greater safety, and reduced manufacturing costs when compared to other technologies. PMID- 8729878 TI - Cytokine receptor expression by solid tumours. AB - The expression of cytokine receptors by a variety of solid tumour tissues was examined, using an immunofluorescence procedure optimized for sensitivity. Several cytokines generally considered as relevant only to the immune and haematopoietic systems were shown to be expressed by solid tumours. For example, breast carcinoma frequently expressed both chains of the IL-3 receptor, the beta chain of the IL-2 receptor, the TNF type two receptor, the signal-transducing chain CD130, and c-kit. The broad expression of cytokine receptors suggests that the receptor profile of individual tumours should be determined before the application of therapy that involves the administration of cytokines. PMID- 8729879 TI - The effector functions of immunoglobulins: implications for therapy. AB - The effector functions of immunoglobulins of the G class (IgGs) are essential for their effective use in therapy. The functions that operate following complex formation with cognate antigen involve binding to C1q (to mediate complement fixation) and the Fc receptors, Fc gamma RI, II and III. Another class of functions that is independent of antigen binding encompasses the transfer of antibodies across the placenta and maintaining the levels in the serum. All effector functions of IgGs are conferred by sequences in the Fc region of antibodies, and this review discusses the localisation of the functions to specific amino acid residues. Such knowledge is of use for the further improvement of IgCs for therapy. PMID- 8729880 TI - Model building of antibody combining sites. AB - The number of high quality crystal structures of antibody fragments available in the Brookhaven Protein Data Bank is rapidly increasing. These structures provide a substantial knowledge base and support model building of novel antibody combining sites. We review some basic principles of antibody structure, describe structure-based modelling procedures, and indicate the strengths and weaknesses of the modelling approach. Applications of antibody models are discussed. PMID- 8729881 TI - Perspectives for the control of tuberculosis. AB - The control of tuberculosis has become more elusive with the increased incidence of HIV, and the continued selection of multidrug-resistant organisms. The intracellular pathogenesis of, and host-response to Mycobacterium tuberculosis present challenges to both classical chemotherapeutic and vaccination approaches, with the organism able to replicate in an unrestricted manner in lung but not other tissues. Adequate control of this pathogen, that has evolved so successfully for its symbiotic exploitation of man, will require complex approaches including additional chemotherapeutics of more acceptable toxicity and efficacy, vaccination and commitment to public health measures. In this review, the worldwide scope of the disease is outlined, and its direct and indirect costs considered. The organism enjoys the protective advantages of a slow replication and of a specialized phagolysosomal intracellular niche, requiring a host response capable of breaching these cellular barriers. The challenges of current vaccine approaches, including live vaccines, target antigen selection and the antigen delivery and adjuventation necessary to elicit adequate pulmonary responses are discussed. Our current understanding is inadequate to control TB and the rekindling of fundamental experimental approaches to the organism, and the host response it evokes, are essential to generate the preventative and therapeutic means necessary for its worldwide control. PMID- 8729882 TI - Genetic and spontaneous models of inflammatory bowel disease in rodents: evidence for abnormalities in mucosal immune regulation. AB - A number of models of spontaneous chronic intestinal inflammation in mice and rats have recently been developed. A characteristic of the majority of these models is that disease developed as a consequence of immune manipulations, suggesting a central role for the immune system in the regulation of intestinal inflammation. Analysis of cytokine patterns in disease showed elevations in TNF alpha and IFN-gamma, characteristic of the T-helper-1 (Th1) pathway, implicating Th1 cells and their cytokines in disease pathogenesis. Strikingly, inflammation did not develop in mice maintained in germ-free conditions, suggesting disease may develop due to a dysregulated inflammatory response to components of the normal flora. Evidence from a number of these models suggests that this potentially pathogenic inflammatory response does not develop in normal animals as it is actively inhibited by a population of CD4+ alpha beta + regulatory T cells and immunosuppressive cytokines such as IL-10 and TGF-beta 1. These new models will allow further investigation into the mechanisms of natural immune regulation and protection in the intestinal tract and how these mechanisms relate to the etiopathogenesis of inflammatory bowel disease (IBD). Furthermore, these models should provide useful insights for the design of effective immunomodulatory therapies for the treatment of IBD in humans. PMID- 8729883 TI - Octreotide acetate in refractory bone marrow transplant-associated diarrhea. AB - OBJECTIVE: To evaluate the effectiveness of octreotide acetate in the treatment of refractory bone marrow transplant-associated diarrhea. DESIGN: Case series encompassing 30 months. SETTING: A 12-bed bone marrow transplant unit at a tertiary care medical center. PARTICIPANTS: Twenty-four patients with bone marrow transplant-associated diarrhea who did not improve with supportive or attapulgite therapy. INTERVENTIONS: Patients received subcutaneous octreotide acetate at doses ranging from 50 to 250 micrograms 2 to 3 times daily. Concurrent treatment with antimotility or antisecretory agents did not occur. MAIN OUTCOME MEASURES: The number of bowel movements and stool volumes were recorded daily. Complete response to octreotide therapy was defined as a reduction of both stool output and stool frequency by more than 50% within 72 hours. Partial response was defined as a reduction of either stool output or stool frequency by more than 50% within 72 hours. Treatment failure occurred if neither of the two parameters decreased by 50% within the designated time period. RESULTS: Twenty-eight treatment challenges were initiated in the 24 patients evaluated. Diarrhea completely or partially subsided in 23 of 28 challenges (82.1%) within 72 hours. Stool output decreased from 1143 +/- 595 at baseline to 252 +/- 356 mL/d within 72 hours (p < 0.005). Stool frequency decreased from a baseline of 7.5 +/- 3.4 to 2.7 +/- 2.2 stools per day within 72 hours (p < 0.005). Adverse effects associated with octreotide were pain or burning at the injection site (24.1%), abdominal pain (13.8%), and increased stool output (6.9%). CONCLUSIONS: These data suggest octreotide acetate significantly reduces stool output and frequency in patients with refractory bone marrow transplant-associated diarrhea. Additional research is necessary before this agent can be recommended for routine use in this patient population. PMID- 8729884 TI - Evaluation of the use of cefuroxime and cefuroxime axetil in an intravenous-oral stepdown program. AB - OBJECTIVE: To characterize cefuroxime and cefuroxime axetil use under the influence of a parenteral-to-oral (iv-po) stepdown program. DESIGN: Open single center retrospective review. SETTING: Tertiary care teaching and referral Canadian hospital with 1100 beds. PATIENTS: A random sample of 78 patients receiving cefuroxime was compared with a random sample of 50 patients receiving iv-po cefuroxime stepdown. RESULTS: During the first 6 months following formulary introduction, 1535 patients received cefuroxime. Stepdown to any oral antibiotic occurred in 22% of patients. Cefuroxime axetil was used as the stepdown agent in 64% of these cases. In a comparison of nonstepdown courses with stepdown courses, some differences were apparent. Nonstepdown treatment courses were primarily prophylactic, whereas stepdown courses were typically initiated as primary therapy for the 10-day management of respiratory tract infections (p < 0.001). Conversion to oral therapy typically occurred on day 5 of parenteral therapy and continued for 5 days. Stepdown was considered possible in 46% of treatment courses in which this process did not happen. When stepdown did occur, it was considered timely in 64% of cases, unnecessarily delayed in 32%, and premature in 4% of treatment courses. Stepdown did not appear to be associated with a negative impact on patient outcome. Mean +/- SD cost of drug therapy per day was less for the stepdown group (US $15.78 +/- $5.97) than the nonstepdown group (US $25.47 +/ $7.87; p < 0.001). CONCLUSIONS: As a result of this study we intend to maintain cefuroxime and cefuroxime axetil on the formulary and continue to judiciously promote the timely conversion to oral therapy. PMID- 8729885 TI - Lorazepam stability in parenteral solutions for continuous intravenous administration. AB - OBJECTIVE: To determine the stability of lorazepam over a 24-hour period when prepared in polyvinyl chloride (PVC) bags at initial concentrations of 0.08 and 0.5 mg/mL. DESIGN: Each concentration was studied at room (21 degrees C) and refrigerator (4 degrees C) temperatures in dextrose 5% (D5W) and NaCl 0.9% solutions. Duplicate test solution admixtures were prepared for each lorazepam concentration, diluent, and temperature. At 0, 1, 4, 8, and 24 hours, duplicate samples were obtained for visual inspection, pH determination, and concentration determination by stability-indicating, reverse-phase HPLC analysis. Compared with baseline, peaks for lorazepam degradation products were not found on any of the study chromatograms. RESULTS: In D5W and NaCl 0.9% solutions, lorazepam loss in excess of 10% by HPLC analysis occurred for concentrations of 0.08 and 0.5 mg/mL at 1 and 4 hours, respectively. CONCLUSIONS: These data suggest that significant loss of lorazepam occurs as the probable result of sorption to PVC bags when admixed in both D5W and NaCl 0.9% solutions at 21 and 4 degrees C. PMID- 8729886 TI - Effect of standard-dose trimethoprim/sulfamethoxazole on the serum potassium concentration in elderly men. AB - OBJECTIVE: To determine the effect of standard-dose trimethoprim/sulfamethoxazole (TMP/SMX) (TMP 160 mg and SMX 800 mg q12h) on the serum potassium concentration. DESIGN: Retrospective and concurrent study. SETTING: A Veterans Affairs Medical Center. PATIENTS: Fifty-three men hospitalized at the Fargo Veterans Affairs Medical Center. Thirty-three patients who received standard-dose TMP/SMX for 3 or more days comprised the study group. Twenty patients who received oral cephradine or amoxicillin for 3 or more days comprised the control group. Patients who received potassium supplements, potassium-sparing diuretics, angiotensin converting enzyme inhibitors, nonsteroidal antiinflammatory drugs, beta-blockers, heparin, known nephrotoxic agents, patients with a serum creatinine concentration of more than 177 mumol/L, and patients with baseline hyperkalemia (serum potassium concentration > 5.1 mmol/L) were excluded. RESULTS: The serum potassium concentration in the study group was 4.22 +/- 0.40 mmol/L and increased by 0.31 +/- 0.38 mmol/L at the end of therapy (p < 0.001). Twenty-six patients in the study group (78.8%) had an increase in the serum potassium concentration during TMP/SMX therapy. Fourteen of these patients had follow-up serum potassium concentrations obtained after completion of therapy. The serum potassium concentration returned to baseline in 10 of these patients. The serum creatinine concentration also increased during therapy. However, the correlation between the increase in the serum potassium concentration and the increase in the serum creatinine concentration was weak (Pearson r = 0.29). The serum potassium in the control group was 4.34 mmol/L and remained essentially unchanged during therapy. CONCLUSIONS: Therapy with standard-dose TMP/SMX is associated with a slight increase in the serum potassium concentration. Routine monitoring of the serum potassium concentration in patients who are treated with standard-dose TMP/SMX therapy is unnecessary. However, TMP/SMX should be considered as a possible cause of unexplained hyperkalemia in elderly patients receiving TMP/SMX therapy. PMID- 8729887 TI - Use of pharmacy computer prescription database to access hypertensive patients for mailed survey research. AB - OBJECTIVE: To evaluate the use of a pharmacy computer prescription database (PCPD) to identify and sample hypertensive patients outside of their treatment setting for mailed, self-administered survey research comparing the presence and lack of participation incentives and mailed follow-up reminders. SETTING: Two independent, privately owned, free-standing community pharmacies. PARTICIPANTS: A total of 735 individuals identified from a PCPD search who had been prescribed at least 1 of 130 PCPD medications potentially used in hypertensive therapy. METHODS: PCPD was searched by the pharmacist/owner per protocol; participants were sent an individually addressed cover letter on pharmacy letterhead signed by the pharmacist that requested voluntary anonymous completion of an enclosed self administered, quality-of-life opinion survey; the first search used no incentive or follow-up; the second search used an incentive and mailed a follow-up reminder. Research protocol followed published ethics guidelines. RESULTS: There was a mean 84% return with incentive and follow-up strategies compared with a mean 25% return without strategies (p < 0.01) for all drug groups between searches; no statistical difference in response was shown between the same drug groups (alpha 1-blockers, calcium-channel blockers, and centrally acting alpha 2 agonists) within searches. CONCLUSIONS: Acceptable response rates (74-93% return) can be obtained with traditional follow-up mailed incentives, prescription issuance within 6 months, incentives to both hypertensive and nonhypertensive responders, and sampling strategies within potential drug groups. Methodology can be adapted to other populations by alternative drug sampling strategies. PMID- 8729888 TI - Multiple delayed peak lithium concentrations following acute intoxication with an extended-release product. AB - OBJECTIVE: To describe delayed peak lithium concentrations after an overdose of extended-release lithium tablets. CASE SUMMARY: A patient with borderline personality disorder and depression ingested extended-release lithium approximately 20.25 g along with other agents. At presentation, the lithium concentration was 1.4 mEq/L. Significant enteral intake was initiated 27 hours after presentation and the lithium concentration 5 hours later increased to 3.2 mEq/L. A second lithium peak concentration of 5.0 mEq/L was noted 40 hours after presentation. Two hemodialysis sessions lasting 4 hours each were performed along with administration of sodium polystyrene sulfonate in sorbitol 20% to enhance lithium elimination and decrease absorption. Eighty-eight hours after presentation, the lithium concentration had decreased to 1.5 mEq/L. DISCUSSION: Delayed and secondary peak lithium concentrations have been reported following an overdose with an extended-release product. Extended-release lithium may form an aggregate in the gastrointestinal tract and/or have delayed absorption secondary to coingested drugs. Toxicity may result if the patient begins enteral intake of drugs, fluids, or nutrition. CONCLUSIONS: Continued monitoring of lithium concentrations after an acute ingestion with an extended-release product are recommended until lithium concentrations are less than 1.5 mEq/L and there are no signs of toxicity, particularly once the patient begins significant enteral intake. PMID- 8729889 TI - Extrapyramidal symptoms following accidental ingestion of risperidone in a child. AB - OBJECTIVE: To describe the development of extrapyramidal symptoms (EPS) precipitated by an accidental overdose of risperidone in a 3.5-year-old boy. CASE SUMMARY: The boy presented to the emergency department with bilateral upward eye gaze, jerky movements of his extremities, and motor restlessness following an accidental ingestion of a single 4-mg risperidone tablet. Decontamination with NaCl 0.9% lavage and activated charcoal with sorbitol was performed. His symptoms responded immediately to intravenous diphenhydramine (on 3 different occasions) during his first 9.5 hours of hospitalization. He experienced no additional EPS, and was discharged home approximately 33 hours following initial presentation. At home, he received three oral doses of diphenhydramine in the 24 hours following hospital discharge because of hand tremor, total body shivering, and eye wandering. These signs resolved without further complications. DISCUSSION: Although the incidence of EPS associated with therapeutic risperidone use is low, its occurrence following overdose is less clearly defined. This represents the first published case, to our knowledge, of risperidone overdose in a child and highlights the potential for dystonic reactions at low doses in this population. Seven intentional overdoses of risperidone in adults (aged 21-68 y) have been reported in the literature and are reviewed. Amounts ingested ranged from 5 to 270 mg. All adult patients appeared to have a relatively benign course. Reported symptoms included drowsiness, slurred speech, altered levels of consciousness, hypertension, tachycardia, electrocardiogram abnormalities, atypical motor behavior, tremors, and other EPS (not specified). CONCLUSIONS: Accidental ingestion of low doses of risperidone can cause EPS in children that may respond well to an anticholinergic agent. Overdose management includes gastrointestinal lavage, activated charcoal with cathartic, cardiovascular monitoring, and supportive therapy. PMID- 8729890 TI - Possible pharmacokinetic interaction with quinidine: ciprofloxacin or metronidazole? AB - OBJECTIVE: To discuss a potential pharmacokinetic interaction between quinidine, ciprofloxacin, and metronidazole. CASE SUMMARY: A 51-year-old black woman was admitted for shortness of breath, abdominal pain, and atrial fibrillation. Procainamide and diltiazem were begun for the atrial fibrillation and ciprofloxacin and metronidazole for suspected diverticulitis. The therapy was switched to quinidine on day 5 because of adverse events associated with procainamide. A trough serum quinidine concentration (SQC) on day 7 was 6.3 micrograms/mL (normal 2-5) with normal QT and QTc intervals. On day 8, the patient was discharged in normal sinus rhythm. She took her last doses of antibiotics on day 15 and a follow-up SQC on day 18 was 2.3 micrograms/mL. DISCUSSION: The possible explanations for the changes in SQCs include: (1) laboratory error, (2) compliance with medication regimen, and (3) altered hepatic metabolism. The first two are not likely in this case. The laboratory verified the elevated SQC and the patient had her prescriptions refilled within appropriate time limits. The third explanation seems more plausible. Quinidine is metabolized by the hepatic mixed-function oxidase system, specifically cytochrome P450 (CYP) 3A4. We found that metronidazole has been shown to inhibit CYP3A activity and ciprofloxacin has been shown to inhibit certain isozymes in the cytochrome P450 system as well. CONCLUSIONS: When metronidazole and ciprofloxacin are administered concomitantly with quinidine, clinicians should be aware of this potential interaction. Quinidine concentrations should be monitored and patients should be assessed for signs and symptoms of quinidine toxicity. PMID- 8729891 TI - Hypersensitivity reactions to etoposide. AB - OBJECTIVE: To report a hypersensitivity reaction to etoposide occurring in a patient after 2 months of drug therapy. CASE SUMMARY: A 20-year-old man with a diagnosis of testicular carcinoma was treated with bleomycin, etoposide, and cisplatin (BEP regimen). After dose 20 of etoposide, an exanthema was noted, which was attributed to etoposide. The patient had received 19 doses of etoposide during the previous 2 months without any sign of an allergic reaction. Rechallenging the patient with etoposide from another batch resulted in recurrence of the exanthema. DISCUSSION: Both etoposide and its excipient (polysorbate 80) are suspected of causing hypersensitivity reactions. Although the exact mechanism of the hypersensitivity reaction is not known, it is believed to be of nonimmunogenic origin. CONCLUSIONS: With a lower rate of infusion of etoposide and/or by premedication with antihistamines and/or corticosteroids, hypersensitivity reactions to etoposide might be prevented in patients with a history of hypersensitivity to this drug. PMID- 8729893 TI - Gaucher's disease: a review. AB - OBJECTIVE: To review the epidemiology, pathophysiology, clinical features, diagnosis, and treatment of Gaucher's disease, focusing on the role of enzyme replacement therapy. DATA SOURCES: A MEDLINE search (from 1984 to July 1995) of English-language literature pertaining to the treatment of Gaucher's disease was performed. Additional references were obtained by reviewing the references of pertinent articles identified through the search. Tertiary sources were also used. STUDY SELECTION AND DATA EXTRACTION: Articles with information on enzyme treatment were selected for review. Articles containing other interesting aspects of the disease or its treatment were also included. DATA SYNTHESIS: Gaucher's disease is the most common lipid storage disorder known and results from a genetic deficiency of the enzyme glucocerebrosidase (glucosylceramidase). Enzyme deficiency results in accumulation of glucocerebroside within the reticuloendothelial system. It may present with hepatosplenomegaly, bone marrow suppression, and bone lesions. The most common of the three subtypes, type 1, is non-neuronopathic. In the rare neuronopathic subtypes, type 2 or 3, there may also be nerve cell destruction within the central nervous system with acute brainstem dysfunction or progressive neurologic deterioration, respectively. In 1991, enzyme treatment became available with the marketing of alglucerase, a placentally derived modified form of glucocerebrosidase. In 1994, a recombinant DNA modified form of glucocerebrosidase, known as imiglucerase, was developed to replace alglucerase. Most published data on enzyme therapy are with alglucerase in patients with type 1 disease. A dosage regimen of 60 units/kg every 2 weeks for moderately to severely ill patients has been effective in reducing hepatosplenomegaly, improving anemia and thrombocytopenia, as well as improving weight gain and growth in children and increasing vigor and self-esteem in adults. Bone involvement is often slow to respond to therapy although pain is frequently improved. Controversy exists as to whether lower dosage regimens are as effective. The role of enzyme therapy in the rarer neuronopathic subtypes remains to be determined, but initial reports have been disappointing. CONCLUSIONS: Enzyme replacement therapy is available for the treatment of type 1 Gaucher's disease, resulting in clinical improvement with enhanced quality of life within the first year of treatment, although improvement in bone disease can take longer. Doses of 60 units/kg every 2 weeks are of clinical benefit to patients with moderate to severe disease. A number of lower dosage regimens have been evaluated in small groups of patients, with satisfactory clinical responses occurring in some of these patients. PMID- 8729892 TI - Aprotinin. AB - OBJECTIVE: To review the clinical pharmacology of aprotinin in patients undergoing surgical procedures involving major blood loss, namely, coronary artery bypass graft (CABG). DATA SOURCES: A MEDLINE search was used to identify French- and English-language publications on aprotinin using the indexing terms aprotinin, cardiothoracic surgery, and hemorrhage. The MEDLINE search was supplemented by review of article bibliographies. Data also were obtained from the approved Canadian and US product labels. STUDY SELECTIONS: All abstracts and uncontrolled and controlled clinical trials were reviewed. DATA EXTRACTION: Study design, population, results, and safety information were retained. Efficacy conclusions were drawn from controlled trials. DATA SYNTHESIS: Aprotinin, a serine protease inhibitor isolated from bovine lung tissue, decreases bleeding after cardiac surgery by mechanisms including antifibrinolytic activity and preservation of platelet function. Several trials have shown that aprotinin reduced blood loss and transfusion requirements in patients undergoing CABG. Its use in other surgical procedures involving major blood loss has been reported. Aprotinin is well tolerated, with minor allergic reactions being the most frequently reported adverse effect. Although unsubstantiated, the possibility that aprotinin could create a prothrombic state leading to early graft occlusion and formation of microthrombi in renal and coronary vasculatures is of concern. CONCLUSIONS: Aprotinin is an effective hemostatic agent in CABG. Clear definitions of indications, dosing, safety, and repeated use remain to be investigated thoroughly. PMID- 8729895 TI - Sotalol prophylaxis and treatment of atrial fibrillation. PMID- 8729894 TI - Exogenous surfactant use in neonates. AB - OBJECTIVE: To review exogenous surfactant use in the treatment of respiratory distress syndrome (RDS) in premature neonates. DATA SOURCES: A MEDLINE search and extensive review of journals was conducted to identify the information for this review from 1983 to 1995 using the following key words: prematurity, surfactant, clinical trials, beractant, Survanta, colfosceril, Exosurf, and neonatal respiratory distress syndrome. STUDY SELECTION AND DATA EXTRACTION: All articles were considered for possible inclusion in the review. Emphasis was placed on controlled studies involving beractant and colfosceril palmitate. DATA SYNTHESIS: Results of published clinical trials using beractant or colfosceril palmitate in premature infants at risk for RDS have shown improvement in the need for supplemental oxygen and ventilatory support over the course of RDS and a reduction in pneumothorax. However, there was little impact on the incidence of intraventricular hemorrhage. Clinical trials studying the treatment of established RDS have also shown similar improvements. In a comparison of prophylactic versus late treatment, no distinct advantage of preventive treatment was found, except among infants less than 26-28 weeks gestation. Single versus multiple-dose studies have shown no specific advantage of more than two doses of colfosceril palmitate. A comparison trial of beractant and colfosceril palmitate has shown no difference in outcome. Long-term follow-up studies have been encouraging among infants being treated with exogenous surfactant. CONCLUSIONS: The use of beractant and colfosceril palmitate in premature infants has clearly decreased morbidity and mortality associated with RDS. Only one trial has compared the efficacy of beractant with that of colfosceril in the treatment of RDS. There does not appear to be a distinct advantage of one product over another. Early treatment of infants at highest risk for RDS, those less than 26 28 weeks gestation, seems to be beneficial over waiting for RDS to progress in severity. Further research needs to be performed to determine the optimal dosing and timing of these agents, as well as comparative trials studying efficacy. Criteria for use of these products need to be further defined to decrease the incidence of unnecessary treatment. PMID- 8729896 TI - Ampicillin-induced maculopapular versus urticarial rash. PMID- 8729897 TI - Albumin prophylaxis of ovarian hyperstimulation syndrome. PMID- 8729898 TI - A perspective on aprotinin. PMID- 8729899 TI - Amphotericin B formulated in a lipid emulsion. PMID- 8729900 TI - Lessons from low enrollment in ACE inhibitor cough study. PMID- 8729901 TI - Severe hyperkalemia with trimethoprim-quinapril. PMID- 8729902 TI - More on comparison of HMG-CoA reductase inhibitors. PMID- 8729903 TI - Comment: dilution of vancomycin for intrathecal or intraventricular administration. PMID- 8729904 TI - Comment: thiazide-induced pulmonary edema. PMID- 8729905 TI - Irradiation of singly and doubly transduced murine neuroblastoma cells expressing B7-1 and producing interferon-gamma reduces their capacity to induce systemic immunity. AB - We have previously reported that immunization with low major histocompatibility complex (MHC) class I expressing murine neuroblastoma (neuro-2a) transduced with B7-1 fails to induce significant protection to wild-type tumor challenge. In this study we investigated whether B7-1 expressing neuro-2a cells can stimulate an effective T-cell response if they were cotransduced with the interferon-gamma (IFN-gamma) gene to upregulate MHC class I. Transfer of both the IFN-gamma and B7 1 genes into neuro-2a (N-2a/B7-1/IFN) almost completely abrogated the tumorigenic potential of this tumor and improved survival when compared with mice receiving the single transductants, N-2a/IFN and N-2a/B7-1. Rejection of N-2a/B7-1/IFN was mediated primarily by CD8+ T cells. When irradiated tumor cells were tested, IFN gamma gene transfer into neuro-2a significantly increased immunogenicity, but transfer of the B7-1 gene did not. However, nonirradiated N-2a/B7-1, N-2a/IFN, and N-2a/B7-1/IFN cells were significantly more effective in eliciting systemic immunity against subsequent wild-type tumor challenge than their irradiated counterparts. N-2a/B7-1/IFN was more immunogenic than N-2a/B7-1 but not more than N-2a/IFN, indicating that B7-1 does not further increase immunogenicity of neuro 2a over that induced by IFN-gamma transduction. These findings should be considered when designing gene modified tumor vaccines for use in human trials. PMID- 8729906 TI - Prodrug-activated gene therapy: involvement of an immunological component in the "bystander effect". AB - The integration and expression of the herpes simplex virus type 1 thymidine kinase (HSV1-TK) gene in localized tumors results in tumor regression after the administration of the specific nucleoside analogue ganciclovir (GCV). Although only 10% to 20% of the tumor cells take up the HSV1-TK gene, the neighboring cells also die, a phenomenon termed "bystander effect.". In the present study, coinjection of the MC26 mouse colon carcinoma cell line and the HSV1-TK expressing retroviral packaging cell line followed after 7 days by the intraperitoneal administration of GCV resulted in almost total tumor regression in the immunocompetent BALB/c mice but not in immunocompromised athymic BALB/c mice. This suggested a strong cell-mediated immune component to the bystander effect. PMID- 8729907 TI - Intracellular antibody against erbB-2 mediates targeted tumor cell eradication by apoptosis. AB - Methods were developed to achieve targeted eradication of the erbB-2 oncoprotein using gene constructs encoding anti-erbB-2 intracellular single-chain antibodies. This method of genetic intervention caused a marked cytocidal effect in erbB-2 overexpressing human ovarian tumor cells. Evaluation of the mechanistic basis of this phenomenon demonstrated that programmed cell death had been induced. Significantly, no cytocidal effect was observed in non-erbB-2-overexpressing tumors. The induction of apoptosis could be shown to be secondary to the intracellular antibody-mediated ectopic localization of the erbB-2 oncoprotein. Thus, the strategy of selective oncogene "knock-out" using intracellular antibodies represents a novel anticancer gene therapy strategy that offers the potential to achieve highly specific, targeted eradication of human tumor cells. PMID- 8729908 TI - Retrovirus-mediated gene therapy of experimental brain neoplasms using the herpes simplex virus-thymidine kinase/ganciclovir paradigm. AB - Recent results in experimental brain tumors indicate that transfer of sensitizing genes to tumor cells in vivo with subsequent drug treatment can reduce tumor masses and prolong the survival of rodents. In the present study, the 9L rat gliosarcoma model was used to evaluate the therapeutic effectiveness of the herpes simplex virus-thymidine kinase (HSV-tk) gene, delivered by a retrovirus vector, against tumor cells in the rat brain after systemic application of the nucleoside analogue ganciclovir (GCV). The HSV-tk gene was inserted into a retroviral vector (pMFG), which was produced using the amphotropic packaging cell line CRIP-MFG-S-HSV-TK. Packaging cells were implanted into established 9L tumors in the brains of syngeneic rats to effect gene delivery to tumor cells, followed by intraperitoneal GCV injections. Treated animals survived significantly longer (more than twice as long) than did the control groups. Brains from GCV-treated and nontreated animals were examined immunohistochemically at different time intervals after grafting of CRIP-MFG-S-HSV-TK cells and GCV treatment. Tumors in GCV-treated animals were significantly smaller as compared with nontreated animals at all time points. Sections stained immunohistochemically for HSV-TK confirmed gene transfer to tumor cells, which could be distinguished from packaging cells by different morphology and immunohistochemical staining for the retroviral envelope protein gp70. Approximately 45% of the cells in tumors implanted with CRIP-MFG-S-HSV-TK cells, but not treated with GCV, showed immunocytochemical staining for HSV-TK, demonstrating a high-efficiency of retrovirus-mediated gene transfer. Tumors in rats treated with packaging cells and GCV showed only 9% HSV-TK-positive cells after treatment, indicating that most cells expressing the HSV-tk gene were killed. The success of this therapeutic modality in experimental animals depends in large parts on the high efficiency of gene delivery and on the immune response against tumor cells. PMID- 8729909 TI - Efficient lipofection with cisplatin-resistant human tumor cells. AB - Seven of seven different cisplatin-resistant human tumor cell lines showed elevated lipofection activity as compared with their sensitive parent cells, although the degree of enhancement was not quantitatively correlated with the degree of cisplatin resistance. Enhanced transfection was seen by using the same reporter gene driven by three different promoter/enhancer sequencer or by using different reporter genes driven by the same promoter/enhancer. Cells resistant to actinomycin D, bleomycin, and nitrogen mustards were not more transfectable than the sensitive parent cells. Although the mechanism of enhanced transfection in cisplatin-resistant cells is not known, data indicated that enhanced transcription, multidrug-resistant phenotype, and methallothionein overexpression do not play a role. PMID- 8729910 TI - Immunogene approach toward cancer therapy using erythrocyte growth factor receptor-mediated gene delivery. AB - In this article we describe an improved method to produce a conjugate of anti erythrocyte growth factor (EGF) receptor monoclonal antibody with polylysine via thio-ether bonds. The resulting antibody/polylysine conjugate was found to be a much more stable DNA (gene) carrier than the previous conjugate formed via disulfide bonds. We designated the conjugate as an "immunoporter" and the immunoporter/DNA (gene) complex as an "immunogene." The fluorescent microscopic observation showed that the immunoporter as well as immunogene bound specifically to the EGF receptors on the cell surface, and the loaded reporter gene, such as beta-galactosidase (beta-GAL), was detected in the cell nucleus at 2 hours after transfection. The enzyme activity from the beta-GAL gene was detected at 12 hours and increased for 3 to 5 days. Similar kinetics were obtained for another reporter gene, chloramphenicol acetyltransferase. Furthermore, the immunoporter delivered the herpes simplex virus thymidine kinase gene and induced substantial suicide effects on tumor cells when gancyclovir or acyclovir was added. Thus, the immunogene approach was successful in delivering therapeutic genes to EGF receptor overexpressing tumor cells. Further technical refinement may prove useful as a supplementary treatment of patients with squamous cell carcinomas. PMID- 8729912 TI - Inhibition of lung cancer proliferation by antisense cyclin D. AB - The growth and tumorigenicity of murine lung cancer cells transfected with an antisense cyclin D1 construct were evaluated in studies pertaining to mouse lung carcinogenesis. This antisense construct inhibited the expression of cyclin D in these cells, significantly reducing both their in vitro proliferation and tumorigenicity in nude mice relative to control cells. These data may have implications regarding the treatment of human neoplasms of aerodigestive tract origin that either overexpress the cyclin D oncogene or exhibit mutations that influence cell cycle progression via cyclin D-dependent mechanisms. PMID- 8729913 TI - Spectral distribution of prosodic information. AB - Prosodic speech cues for rhythm, stress, and intonation are related primarily to variations in intensity, duration, and fundamental frequency. Because these cues make use of temporal properties of the speech waveform they are likely to be represented broadly across the speech spectrum. In order to determine the relative importance of different frequency regions for the recognition of prosodic cues, identification of four prosodic features, syllable number, syllabic stress, sentence intonation, and phrase boundary location, was evaluated under six filter conditions spanning the range from 200-6100 Hz. Each filter condition had equal articulation index (AI) weights, AI = 0.01; p(C)isolated words approximately equal to 0.40. Results obtained with normally hearing subjects showed that there was an interaction between filter condition and the identification of specific prosodic features. For example, information from high frequency regions of speech was particularly useful in the identification of syllable number and stress, whereas information from low-frequency regions was helpful in identifying intonation patterns. In spite of these spectral differences, overall listeners performed remarkably well in identifying prosodic patterns, although individual differences were apparent. For some subjects, equivalent levels of performance across the six filter conditions were achieved. These results are discussed in relation to auditory and auditory-visual speech recognition. PMID- 8729911 TI - Adenovirus-mediated p53 gene transfer inhibits growth of human tumor cells expressing mutant p53 protein. AB - Human malignancies are often characterized by mutations of the p53 tumor suppressor gene. In a large proportion of cases, the mutation results in production of an altered protein that can bind and inactivate the wild-type gene product. This "dominant-negative" activity of mutant p53 molecules may limit the utility of p53 gene therapy of cancer. Using replication-deficient recombinant adenoviruses (rAd-p53) as a p53 gene delivery system, we evaluated the effects of p53 reintroduction on a series of 45 human cell lines containing wild-type, mutated, or no p53 protein. Results indicate a p53-specific, dose-dependent, and promoter-specific growth inhibition of a majority of p53-altered cell lines that correlates with the degree of adenovirus transgene expression. Similar effects were not observed on cells containing wild-type p53. rAd-p53 inhibited the growth of cells expressing various mutant p53 proteins including those characterized as "dominant negative mutants", and the antiproliferative effects were not abrogated by high levels of endogenous mutated p53 protein. In vivo, rAd-p53 also suppressed tumor growth and increased survival of nude mice bearing tumors that express mutant p53. These results support a role for p53 gene therapy of cancer, including malignancies harboring mutations in this tumor suppressor gene. PMID- 8729914 TI - Peaks in the frequency response of hearing aids: evaluation of the effects on speech intelligibility and sound quality. AB - In a series of experiments, we introduced peaks of 10, 20, and 30 dB, in various combinations, onto a smooth reference frequency response. For each of the conditions, we evaluated speech intelligibility in noise, using a test as developed by Plomp and Mimpen (1979), and sound quality (for both speech and music), using a rating-scale procedure. We performed the experiments with 26 listeners with sensorineurally impaired hearing and 10 listeners with normal hearing. Signal processing was accomplished digitally; for each listener, the stimuli were filtered and subsequently amplified so that the average speech spectrum was well above the threshold of hearing at all frequencies. The results show that, as a result of the introduction of peaks onto the frequency response, speech intelligibility is affected more for the listeners with impaired hearing than for those with normal hearing. Sound-quality judgments tend to be less different between the listener groups. Conditions with 30-dB peaks especially show serious effects on both speech intelligibility and sound quality. PMID- 8729915 TI - Noise exposure associated with hearing aid use in industry. AB - Although noise may be innocuous in many vocational environments, there is a growing concern in industry that it can reach hazardous levels when amplified by hearing aids. This study examined the daily noise exposures associated with hearing aid use in industry. This was done by both laboratory and site measurements in which hearing aids were coupled to the microphone of an integrating sound level meter or dosimeter. The former method involved the use of recorded railroad and manufacturing noise and a Bruel and Kjaer 4128 Head and Torso simulator. In the latter procedure, a worker wore one of three hearing aids coupled to a dosimeter during 8-hour shifts in a manufacturing plant. Both methods demonstrated that even when amplified by mild-gain hearing aids, noise exposures rose from time-weighted averages near 80 dBA to well above the OSHA maximum of 90 dBA. The OSHA maximum was also exceeded when moderate and high gain instruments were worn in non-occupational listening environments. The results suggest that current OSHA regulations that limit noise exposure in sound field are inappropriate for hearing aid users. PMID- 8729916 TI - A within-subject comparison of adult patients using the Nucleus F0F1F2 and F0F1F2B3B4B5 speech processing strategies. AB - This study compares the Nucleus F0F1F2 and F0F1F2B3B4B5 (also known as "Multipeak") of "Mpeak") processing schemes in 17 patients wearing the Mini Speech Processor. All patients had at least 18 months implant experience using the F0F1F2 processing strategy. For this study, they were switched to the F0F1F2B3B4B5 processing strategy for 3 months. They then returned to using the F0F1F2 strategy for 3 months, then used the F0F1F2B3B4B5 strategy again for 3 months, and lastly used the F0F1F2 strategy for 3 months. Performance' was evaluated with both schemes after each interval, using speech recognition tests and subjective ratings. Overall, differences between the results for the two processing schemes were not large. Average performance was somewhat better for the F0F1F2B3B4B5 strategy for word and sentence identification, but not for any of the other speech measures. Superior performance was observed in 8 patients with the F0F1F2B3B4B5 strategy. However, 6 of the 8 individuals were significantly better on only one of the six speech measures in the test battery. The other 2 patients performed better on two of the speech measures. Superior performance was also observed in 3 patients with F0F1F2 strategy for consonant recognition. For the remaining patients, there was little difference in their performance with the two strategies. Information transmission analyses indicated that the F0F1F2B3B4B5 strategy transmitted consonant duration and frication cues more efficiently than F0F1F2. Experience with one strategy appeared to benefit performance with the other strategy. PMID- 8729917 TI - Discriminability and perceptual weighting of some acoustic cues to speech perception by 3-year-olds. AB - Studies of children's speech perception have shown that young children process speech signals differently than adults. Specifically, the relative contributions made by various acoustic parameters to some linguistic decisions seem to differ for children and adults. Such findings have led to the hypothesis that there is a developmental shift in the perceptual weighting of acoustic parameters that results from experience with a native language (i.e., the Developmental Weighting Shift). This developmental shift eventually leads the child to adopt the optimal perceptual weighting strategy for the native language being learned (i.e., one that allows the listener to make accurate decisions about the phonemic structure or his or her native language). Although this proposal has intuitive appeal, there is at least one serious challenge that can be leveled against it: Perhaps age-related differences in speech perception can appropriately be explained by age-related differences in basic auditory-processing abilities. That is, perhaps children are not as sensitive as adults to subtle differences in acoustic structure and so make linguistic decisions based on the acoustic information that is most perceptually salient. The present study tested this hypothesis for the acoustic cues relevant to fricative identity in fricative-vowel syllables. Results indicated that 3-year-olds were not as sensitive to changes in these acoustic cues as adults are, but that these age-related differences in auditory sensitivity could not entirely account for age-related differences in perceptual weighting strategies. PMID- 8729918 TI - Time-interval measurement of stuttering: establishing and modifying judgment accuracy. AB - The purpose of this study was to determine whether accuracy training for interval judgments of stuttering might generalize to increased accuracy and/or interjudge agreement for intervals other than those used during training. Ten upper-division speech-language pathology students judged 5-s audiovisually recorded speech intervals as stuttered or nonstuttered in a series of group and single-subject experiments. Judgment accuracy was determined with respect to judgments provided previously by 10 recognized authorities on stuttering and its treatment. Training occurred within single-subject experiments that used multiple baselines across speakers and repeated generalization probes to assess training effects. Results showed that judgment accuracy tended to increase after training for speakers used during the training process as well as for unfamiliar speakers. Results also replicated previous findings of slight increases in interjudge and intrajudge agreement after interval-judgment training. The implications of these results for developing a valid and reliable stuttering measurement system are discussed. PMID- 8729919 TI - Acoustic correlates of breathy vocal quality: dysphonic voices and continuous speech. AB - In an earlier study, we evaluated the effectiveness of several acoustic measures in predicting breathiness ratings for sustained vowels spoken by nonpathological talkers who were asked to produce nonbreathy, moderately breathy, and very breathy phonation (Hillenbrand, Cleveland, & Erickson, 1994). The purpose of the present study was to extend these results to speakers with laryngeal pathologies and to conduct tests using connected speech in addition to sustained vowels. Breathiness ratings were obtained from a sustained vowel and a 12-word sentence spoken by 20 pathological and 5 nonpathological talkers. Acoustic measures were made of (a) signal periodicity, (b) first harmonic amplitude, and (c) spectral tilt. For the sustained vowels, a frequency domain measure of periodicity provided the most accurate predictions of perceived breathiness, accounting for 92% of the variance in breathiness ratings. The relative amplitude of the first harmonic and two measures of spectral tilt correlated moderately with breathiness ratings. For the sentences, both signal periodicity and spectral tilt provided accurate predictions of breathiness ratings, accounting for 70%-85% of the variance. PMID- 8729920 TI - Glottal airflow characteristics of women's voice production along an aging continuum. AB - Anatomical and physiological changes of the speech production mechanism that occur with aging may result in phonatory distinctions between older and younger speakers. This investigation examined amplitude-based glottal airflow characteristics from sustained vowel production in healthy adult women. Sixty women participated in this study, 10 each in six age groups of 20-, 30-, 40-, 50 , 60-, and 70-year-olds. Measures included peak, alternating, and minimum glottal airflow. Additionally, a ratio of minimum to peak glottal airflow was calculated. Results from an analysis of variance indicated no significant group mean difference for any of the dependent measures. A greater variability in peak glottal airflow for the 70-year-old age group as compared to the 20-year-old age group was found. None of the dependent variables were significantly related to age and therefore were not good predictors of age. The results imply that laryngeal senescence in healthy women may not be significant enough to affect the magnitude of phonatory function parameters. Either the assumed anatomical changes produce less significant phonatory change in the healthy individual or the healthy individual is more capable of using strategies to counteract degenerative laryngeal changes. PMID- 8729921 TI - Activity of intrinsic laryngeal muscles in fluent and disfluent speech. AB - The goal of the present experiment was to determine if stuttering is associated with unusually high levels of activity in laryngeal muscles. Qualitative and quantitative analyses of thyroarytenoid and cricothyroid recordings from 4 stuttering and 3 nonstuttering adults revealed the following: Compared to periods of fluent speech, intervals of disfluent speech are not typically characterized by higher levels of activity in these muscles; and when EMG levels during conversational speech are compared to maximal activation levels for these muscles (e.g., those observed during singing and the Valsalva maneuver), normally fluent adults show robust and sometimes near maximal recruitment during conversational speech. The adults who stutter had a lower operating range for these muscles during conversational speech, and their disfluencies did not produce relatively high activation levels. In summary, the present data require us to reject the claim that adults with a history of chronic stuttering routinely produce excessive levels of intrinsic laryngeal muscle activity. These results suggest that the use of botulinum toxin injections into the vocal folds to treat stuttering should be questioned. PMID- 8729922 TI - Stuttering and phonological disorders in children: examination of the Covert Repair Hypothesis. AB - The purpose of this study was to evaluate whether the Covert Repair Hypothesis (CRH; Postma & Kolk, 1993), a theory designed to account for the occurrence of speech disfluencies in adults who stutter, can also account for selected speech characteristics of children who stutter and demonstrate disordered phonology. Subjects were 9 boys who stutter and exhibit normal phonology (S + NP; mean age = 61.33 months; SD = 10.16 months) and 9 boys who stutter and exhibit disordered phonology (S + DP; mean age = 59.11 months; SD = 9.37 months). Selected aspects of each child's speech fluency and phonology were analyzed on the basis of an audio/videotaped picture-naming task and a 30-min conversational interaction with his mother. Results indicated that S + NP and S + DP children are generally comparable in terms of their basic speech disfluency, nonsystematic speech error, and self-repair behaviors. CRH predictions that utterances produced with faster articulatory speaking rates or shorter response time latencies are more likely to contain speech errors or speech disfluencies were not supported. CRH predictions regarding the co-occurrence of speech disfluencies and speech errors were supported for nonsystematic ("slip-of-the-tongue"), but not for systematic (phonological process/rule-bases), speech errors. Furthermore, neither S + NP nor S + DP subjects repaired their systematic speech errors during conversational speech, suggesting that systematic deviations from adult forms may not represent true "errors, " at least for some children exhibiting phonological processes. Findings suggest that speech disfluencies may not represent by-products of self repairs of systematic speech errors produced during conversational speech, but that self-repairs of nonsystematic speech errors may be related to children's production of speech disfluencies. PMID- 8729923 TI - The equilibrium point hypothesis and its application to speech motor control. AB - In this paper, we address a number of issues in speech research in the context of the equilibrium point hypothesis of motor control. The hypothesis suggests that movements arise from shifts in the equilibrium position of the limb or the speech articulator. The equilibrium is a consequence of the interaction of central neural commands, reflex mechanisms, muscle properties, and external loads, but it is under the control of central neural commands. These commands act to shift the equilibrium via centrally specified signals acting at the level of the motoneurone (MN) pool. In the context of a model of sagittal plane jaw and hyoid motion based on the lambda version of the equilibrium point hypothesis, we consider the implications of this hypothesis for the notion of articulatory targets. We suggest that simple linear control signals may underlie smooth articulatory trajectories. We explore as well the phenomenon of intraarticulator coarticulation in jaw movement. We suggest that even when no account is taken of upcoming context, that apparent anticipatory changes in movement amplitude and duration may arise due to dynamics. We also present a number of simulations that show in different ways how variability in measured kinematics can arise in spite of constant magnitude speech control signals. PMID- 8729924 TI - How children learn to organize their speech gestures: further evidence from fricative-vowel syllables. AB - Previous studies with fricative-vowel (FV) syllables have shown that the difference in overall spectrum between fricatives is less in children's speech than in that of adults, but that fricative noises show greater differences in the region of the second formant (F2) as a function of the upcoming vowel than those of adults at corresponding points in the fricative. These results have been interpreted as evidence that children produce fricatives that are not spatially differentiated as those of adults and that children initiate vowel gestures earlier during syllable production than adults do (Nittrouer, Studdert-Kennedy, & McGowan, 1989). The goals of the present study were (a) to replicate the previous age-related difference for F2 with FV syllables; (b) to test the alternative interpretation that age-related differences in fricative f2 reflect age-related differences in vocal-tract geometry; (c) to determine whether age-related differences in F2 (and so, by inference, in articulatory organization) might extend beyond the syllable boundaries, perhaps into the schwa of a preceding unstressed syllable; and (d) determine if gestures other than fricative gestures show less spatial differentiation in children's than in adults' speech. To these ends, F2 frequencies were measured in schwa-fricative-vowel utterances (consisting of the fricatives /s/ and [symbol:see text] and of the vowels /i/ and /a/) from 40 speakers (10 each of the ages of 3, 5, 7 years, and adults) at three locations (for the entire schwa, for 10 ms of fricative noise centered at 30 ms before voicing onset, and 10 pitch periods from vocalic center). Results of several analyses supported four conclusions: (a) the earlier finding was replicated; (b) age-related differences in vocal-tract geometry could not explain the age-related difference in vowel effects on fricative noise; (c) children master intersyllabic gestural organization prior to intrasyllabic gestural organization; and (d) unlike fricative gestures, children's vowel gestures are more spatially distinct than those of adults. PMID- 8729925 TI - Creating filters with arbitrary response characteristics for use in hearing and speech research. AB - Digital filters with conventional lowpass, highpass, bandpass, and band reject frequency response curves are perfectly adequate for many research applications in speech and hearing. However, there are some specialized applications for which these conventional response characteristics are less than ideal. In this paper we describe a simple method for generating digital filters with virtually any amplitude and phase response. The process involves (a) calculating the impulse response of a finite impulse response filter from a text file that specifies the desired magnitude and phase response of the filter, and (b) convolving the impulse response with the input signal. Sample applications of this method are described. PMID- 8729926 TI - Fluency effect of frequency alterations of plus/minus one-half and one-quarter octave shifts in auditory feedback of people who stutter. AB - The effect of frequency alterations in auditory feedback of people who stutter on stuttering frequency was investigated. Twelve participants who stutter read aloud under nonaltered auditory feedback (NAF) and four conditions of frequency-altered feedback ([FAF], plus/minus one-half and one-quarter octaves) at normal and fast speech rates. Stuttering frequency was significantly higher while reading aloud with NAF relative to the four conditions of FAF (p < 0.05). There were no differences among participants' stuttering frequency between the four FAF conditions (p > 0.05). Reductions in stuttering frequency of approximately 50% to 60% were found with FAF relative to NAF. More disfluencies occurred with the fast versus the normal speech rate condition (p = .0007) irrespective of auditory feedback condition. These findings suggest that slight alterations in the frequency of auditory feedback of people who stutter are fluency-enhancing. PMID- 8729927 TI - Applications of disfluencies in measurements of stuttering. PMID- 8729928 TI - The problem of generalizing to a language population: a "random" controversy. AB - The rationale behind the use of item statistics as well as objections to their use are reviewed. It is argued that investigators conducting language research with disordered populations should follow the example of those conducting language research with normal populations and consider calculating item statistics if they wish their results to generalize beyond the specific items tested. PMID- 8729929 TI - Assessment information for predicting upcoming change in language production. AB - Initial language assessments are used not only to determine the presence of a language problem and establish eligibility for intervention, but also to provide information about a child's readiness for immediate change in language growth. This study explored static assessment profiling (specific variables and discrepancies in performance) and dynamic assessment results to determine their relative effectiveness for predicting immediate change. Correlation data were used to examine how well each assessment measure predicted upcoming language production changes for children with specific expressive language impairment. Results indicated that dynamic assessment outcomes were most highly correlated with immediate language growth, followed by discrepancy in receptive and expressive language age. Findings are discussed in terms of their clinical and theoretical importance. PMID- 8729930 TI - Consonant-vowel interdependencies in babbling and early words: preliminary examination of a locus equation approach. AB - Consonant-vowel productions at two distinct stages of language development were studied in a single female child. At 12 months canonical babbling syllables (N = 144) identified by a panel of listeners as comprising [bV], [dV], and [gv] tokens were acoustically analyzed by measuring F2 transition onset and F2 midvowel frequencies and plotting their relationship as locus equations for each stop category. A regression analysis performed on these scatterplots revealed differential slopes and y-intercepts as a function of stop place. The same analysis was performed 9 months later on CV utterances (N = 243) produced as syllable-initial segments of real words by the same child. Whereas labial and velar locus equation parameters moved toward more adult-like values, alveolar slope and y-intercept moved away from adult values and more in the direction of decreased coarticulation between vowel and consonant. There was greater scatter of data points around the regression line for production of words compared to babbling. These results are compared to locus equations obtained from 3-5-year olds and adults. Locus equations appear to be useful as an empirical developmental probe to document how CV productions gradually approach adult categorical standards. PMID- 8729931 TI - Persistence of non-standard dialect in school-age children. AB - This study investigated the persistence of non-standard dialect production among 114 African American and White children in grades 3, 5, and 7. A dialect shift premise suggests that a large and uniform decline in dialectal features occurs in the language of school-age children. Three experimental tasks were administered. The results indicated that dialect awareness and discrimination increased as grade in school increased; a dialect shift occurred between grades 3 and 5; non standard dialect production and comprehension of standard dialect were not associated; and that there was no difference in non-standard dialect production among African American and White students. PMID- 8729932 TI - Idiom understanding in Australian youth: a cross-cultural comparison. AB - In this developmental study, idiom understanding was examined in Australian students from Grades 5 and 8 (n = 50 per group; mean ages = 10.7 and 13.8, respectively). Twenty-four idioms with familiarity ratings ranging from high to low (as judged by Australian adolescents) were each presented in a brief story context. The students read each story and selected the best interpretation of the idiom from a set of four answer choices. Results indicated that performance on the task improved as a function of increasing grade level and that idiom familiarity was significantly correlated to idiom understanding for both groups of students. These results, which were consistent with a previous study of American students of comparable educational levels (Nippold & Taylor, 1995), provide further support for the "language experience" hypothesis of figurative language development. In replicating the previous developmental study, evidence of external validity is provided. PMID- 8729933 TI - Length of estrous cycle and gestation in the Arabian oryx (Oryx leucoryx) and the importance of the male presence for induction of postpartum estrus. AB - The reproduction of the Arabian oryx (Oryx leucoryx) has been studied in a breeding center in Taif (Saudi Arabia). In the first experiment, three groups of females were assigned to groups immediately after calving. In group 1, from the day of calving the females had visual and olfactory but not physical contact with a male. The calves were removed on the day of birth and each female was placed with a male on Day 45. In group 2, females and their calves were placed individually with a male the day after calving. In group 3, females and their calves were kept with neither visual nor olfactory contact with a male and then placed with a male on Day 90. The estrous cycle was deduced from assays of plasma progesterone up to Day 120 after calving. All females exhibited a postpartum estrus a few days after parturition but the occurrence of the estrus was significantly delayed in females of group 3. The length of the estrous cycle was 23.7 +/- 1.3 days and the gestation length was 255.2 +/- 1.5 days. In the second experiment, a group of adult females was formed in 1987 and the intervals between calving recorded over a period of 5 years. The mean interval between calving was 285 +/- 2 days (n = 99 gestations). However, 66.6% of these intervals averaged 276 days whereas 19.2% had averaged 294 and 13% averaged 321 days. PMID- 8729934 TI - Interrelationship of day length and temperature on the control of gonadal development, body mass, and fat score in white-crowned sparrows, Zonotrichia leucophrys gambelii. AB - We tested the effects of naturally relevant environmental temperatures on long day-induced reproductive development in male and female white-crowned sparrows, Zonotrichia leucophrys gambelii. Transfer from short days (8L 16D) to long days (20L 4D) resulted in rapid testicular development and partial ovarian development as has been reported many times previously. Exposure of experimental groups to low (5 degrees), moderate (20 degrees), and high (30 degrees) temperature during photostimulation had only subtle effects on plasma levels of follicle-stimulating hormone and luteinizing hormone over time and no effects on the size of testes, cloacal protuberance, ovaries, or brood patch at Day 30 of treatment. Long days resulted in the well known increase in body mass and fat score, indicative of preparations for migration. In females, treatment with low temperature resulted in a reduction in the premigratory increase in fat and body mass when transferred to long days. This was accompanied by an increase in plasma levels of corticosterone during the early stages of photostimulation at low temperature. Temperature regimes had no effects on fattening or body mass in males, despite an early increase in plasma corticosterone at low temperature. Circulating levels of thyroxine (T4) and triiodothyronine (T3) increased to varying degrees following photostimulation. Temperature treatment had no effect on plasma levels of thyroid hormones in males, but low temperature did inhibit thyroid hormone secretion (particularly T4) in females. Although reproductive development appears to be resistant to naturally relevant temperature extremes in both sexes, low environmental temperature impaired preparations for migration in females but not males. This effect may be mediated through glucocorticosteroids and not thyroid hormones. Reasons for the sexual dimorphism in this response are unknown, but may be related to sexual selection for males to arrive on the breeding grounds ahead of females regardless of local weather conditions. PMID- 8729935 TI - In vitro biosynthesis of androgens in the Australian lungfish, Neoceratodus forsteri. AB - The serum concentration of testosterone was estimated from a population of wild lungfish over 6-7 years of sampling. Male lungfish were found to have high circulating levels of testosterone (approximately 50 ng/ml) which varied seasonally and could be correlated with spermatogenesis as judged by testis histology. Incubation of testis tissue slices with [3H]progesterone, [3H]17 hydroxyprogesterone, or [3H]testosterone confirmed that testosterone is the major androgen in Neoceratodus. Not even trace amounts of 11-keto- or 11 beta hydroxytestosterone or 5 alpha-dihydrotestosterone could be identified by TLC separations. There was little or no conjugation of steroids by the testes, except during the spawning season, when glucuronides of androstenedione and testosterone were produced. PMID- 8729936 TI - Somatotropin release-inhibiting factor and galanin innervation in the hypothalamus and pituitary of seabream (Sparus aurata). AB - The distribution of galanin (GAL) and somatotropin-release-inhibiting-factor (SRIF) immunoreactivity in the hypothalamus and pituitary of the sea bream (Sparus aurata) was studied by immunocytochemistry. An extensive system of neurons immunoreactive with antisera to the two peptides was identified throughout the brain with staining particularly in the hypothalamus. In the hypothalamus, GAL immunoreactive perikarya were detected principally in the nucleus preopticus and nucleus tuberis. Major nerve tracts were observed to sweep down from the hypothalamic nuclei and reached the pituitary via the preoptico hypophysial tract. Many of the fibers had varicose swellings indicating they were secretory. SRIF immunoreactivity was distributed similarly to GAL but the network of nerve fibers was less dense; no colocalization of these two peptides was seen. SRIF immunoreactive perikarya were present in the preoptic nucleus, the tuberal nucleus, and the basolateral hypothalamus. These perikarya were large and densely staining and were predominately bipolar, although some multipolar perikarya were observed. In the pituitary GAL and SRIF immunoreactivities were confined principally to the pars distalis where fibers infiltrated between growth hormone, prolactin, and adrenocorticotrophic cells. More of the fibers were immunoreactive for SRIF than for GAL. There was no immunoreaction for GAL or SRIF in any of the pituitary cells. There is thus morphological evidence for a neuroendocrine control of the pars distalis by GAL and SRIF and for a possible functional interaction between these two systems. PMID- 8729937 TI - Changes of thyrotropin-releasing hormone concentration in the brain and levels of prolactin and thyroxin in the serum during spawning migration of the chum salmon Oncorhynchus keta. AB - Changes in concentration of thyrotropin-releasing hormone (TRH) in various regions of the brain of female and male chum salmon Oncorhynchus keta during their spawning migration toward the home stream were examined to determine the physiological role of TRH. In the same specimens, serum levels of prolactin (PRL) and thyroxin (T4) were also examined and compared with the change of TRH concentration. TRH concentration in the olfactory bulb of males increased significantly just before the upstream migration or entering the river and dropped after entering the home stream. TRH concentration in the olfactory lobe was also slightly higher in Class 2 salmons with nuptial coloration captured in coastal water than in the other groups. TRH concentrations in hypothalamus and pituitary remained relatively low during the spawning migration. The change of TRH concentration in females was similar to that in males, but was smaller and with a wider range. The serum level of PRL was extremely low in both males and females collected in coastal waters (Classes 1 and 2) and increased after the fish entered the river. The serum level of T4 in both males and females tended to be low, as did the TRH concentration in the hypothalamus and pituitary. Immunohistochemistry of the brain using anti-proTRH serum revealed many proTRH cell bodies in the olfactory bulb and olfactory lobe. These results indicate that TRH in the homing salmon is produced mainly in the olfactory bulb and lobe and that its production may be related to changes in olfactory function during migration. PMID- 8729939 TI - Identification and androgen regulation of a 156-kDa hemeprotein in the harderian gland of the Syrian hamster. AB - An abundant cytosolic protein from hamster Harderian glands was partially purified by ultracentrifugation and analyzed on SDS-PAGE. This 156-kDa protein occurs in male but not in female glands. Castration of adult males for different time periods resulted in a gradual disappearance of the protein over 5 weeks. Daily administration of testosterone, 5 alpha-dihydrotestosterone, 3 alpha- or 3 beta-androstanediol (0.5 mg/day/15 days), to castrated males maintained the levels of the protein. In females, the protein was induced with the same compounds but not with 17 beta-estradiol. These observations indicate an androgen mediated hormonal regulation. In developing animals, the 156-kDa protein was detectable in glands of 2-week-old males and females; it was not detected in 4 week-old females. Its specific reactions to 3,3',5,5'-benzidine derivatives strongly suggest heme groups associated with this Harderian gland protein. PMID- 8729938 TI - Expression of salmon gonadotropin-releasing hormone (GnRH) and chicken GnRH-II precursor messenger ribonucleic acids in the brain and ovary of goldfish. AB - The complementary DNAs (cDNA) encoding the [Trp7Leu8]gonadotropin-releasing hormone (salmon GnRH; sGnRH) precursor and the [His5Trp7Tyr8]GnRH (chicken GnRH II; cGnRH-II) precursor of the goldfish brain were isolated and sequenced using reverse transcription and rapid amplification of cDNA ends (RACE). The sGnRH precursor cDNA consists of 540 bp, including an open reading frame of 282 bp, and the cGnRH-II precursor cDNA consists of 682 bp, including an open reading frame of 258 bp. The 94 amino acid-long goldfish sGnRH precursor and 86 amino acid-long goldfish cGnRH-II precursor have the same molecular architecture as GnRH precursors identified to date in other vertebrate species. Using two sets of primers designed to be sense and antisense to the goldfish brain sGnRH precursor cDNA sequence, reverse transcription-polymerase chain reaction (RT-PCR) amplification of total RNA from brain and ovary at gonadal recrudescent, mature ( = prespawning), and postovulatory stages resulted in two predicted sizes of PCR products. The intensities of staining signals of ethidium bromide were similar between brain and ovary samples. The same RT-PCRs were carried out with two sets of primers for cGnRH-II precursor cDNA, resulting in two PCR products of predicted size; however, the ethidium bromide staining signals are much weaker for products amplified from ovarian cDNA than that from brain cDNA. Restriction enzyme analysis verified the expected RT-PCR products. Sequence analysis of ovarian sGnRH precursor cDNA generated by RACE of total RNA from recrudescent ovarian tissue revealed the identical sequence to that of the brain sGnRH cDNA. Northern blot analysis detected a single mRNA transcript of approximately 650 bases for the sGnRH precursor in both the brain and ovary, and 750 bases for the cGnRH-II precursor in the brain. These results demonstrate that two forms of GnRH precursor (sGnRH and cGnRH-II) mRNA are expressed in goldfish brain tissue and that the sGnRH transcript and a low level of cGnRH-II transcript are also expressed in the goldfish ovary. PMID- 8729940 TI - Comparison of the ontogenesis of thyroid hormones, growth hormone, and insulin like growth factor-I in ad libitum and food-restricted (altricial) European starlings and (precocial) Japanese quail. AB - In this study, we compare the ontogenic patterns for thyroid hormones, growth hormone (GH), and insulin-like growth factor-I (IGF-I) in altricial European starlings and precocial Japanese quail and examine the effects of feed restriction on these species. The most marked difference in development between the altricial and precocial birds was with respect to plasma thyroid hormone patterns. In the starling, circulating concentrations of triiodothyronine (T3) and thyroxine (T4) were very low in embryos, then increased progressively after hatching to peak at 10-11 days of age. In contrast, in quail, in which other studies have shown that most thyroid maturation occurs during the embryonic and peri-hatch periods, the circulating concentrations of T3 and T4 showed little posthatch ontogenic change. Plasma concentrations of both GH and IGF-I showed similar patterns in both species with a posthatch rise (peak at 3 days in starlings and 8 days in quail), followed by a decline. Food restriction to maintain body weight resulted in decreased plasma concentrations of T3 and IGF-I in both species. After return to ad libitum feeding, plasma T3 and IGF-I increased in both early and late restricted starlings and in late restricted quail. Although both species responded to food restriction with similar patterns of endocrine change, age-related differences in the magnitude of hormonal responses were observed. PMID- 8729941 TI - Circulating levels of prolactin and progesterone in a wild population of red kangaroos (Macropus rufus) Marsupialia: Macropodidae. AB - Circulating progesterone and prolactin levels were measured in shot and live caught wild red kangaroos using radioimmunoassays validated for the red kangaroo. The objective of the study was to correlate hormone profiles with reproductive status and determine if red kangaroos follow the general pattern elucidated for other macropodids. During Phase 2a lactation ( < 70 days) plasma progesterone concentrations were < 189 pg/ml (n = 41). This value increased to > 600 pg/ml (n = 32) during the transition to Phase 3 lactation (181 to 235 days) when the quiescent corpus luteum and embryo were reactivated. Progesterone concentrations then decreased to < 300 pg/ml (n = 29) during dual lactation when females were suckling a neonate and a young at foot. Concentrations of prolactin during Phase 2a were < 6 ng/ml (n = 17). Coincident with the period of reactivation of the diapausing blastocyst (181 to 235 days), plasma prolactin concentrations increased to 15 ng/ml (n = 32), then decreased and remained low through the subsequent stage of dual lactation. These results indicate that progesterone and prolactin profiles in wild red kangaroos follow patterns found previously in other macropodid species, the tammar and Bennett's wallabies. PMID- 8729942 TI - Thyroid hormone deiodination pathways in brain and liver of rainbow trout, Oncorhynchus mykiss. AB - Outer-ring (5') deiodination (ORD) and inner-ring (5) deiodination (IRD) of L thyroxine (T4) and 3,5,3'-triiodo-L-thyronine (T3) were studied in whole-brain microsomes of rainbow trout and compared with liver deiodination. Brain T4ORD activity (apparent Km = 1.2-2.5 nM; V(max) = 0.10-0.14 pmol/hr/mg microsomal protein) was less than T4IRD activity (apparent Km = 4.9; V(max) = 0.32) and T3IRD activity (apparent Km = 5.2-5.4; V(max) = 1.1-2.0); T3ORD activity was negligible. All three brain deiodinase pathways shared the following properties: pH optima between 7.0 and 7.3, activity enhanced by dithiothreitol (10 mM), weak inhibition by 6-n-propyl-2-thiouracil and iodoacetate, but stronger inhibition by aurothioglucose. Based on competitive inhibition, the substrate preference for brain T4ORD was T4 = tetraiodothyroacetic acid (TETRAC) > 3,3',5'-triiodo-L thyronine (rT3) > 3,5,3'-triiodothyroacetic acid (TRIAC) >> T3 > 3,5-diiodo-L thyronine (3,5-T2). A comparable substrate preference profile was obtained for liver T4ORD (Km 1 nM). Both T4IRD and T3IRD in brain had similar substrate preference profiles (rT3 > 3,5-T2 > T4 > T3) which differed from that of T4ORD. Negligible T4IRD and T3IRD activities existed in liver. We conclude that for rainbow trout (i) T4ORD systems in brain and liver are similar, and consistent with a common enzyme that does not match exactly either mammalian type I or II deiodinases, (ii) brain T4IRD and T3IRD enzymes share several common properties, and correspond functionally to the mammalian type III deiodinase, and (iii) under normal physiological conditions the predominant deiodinase pathways in brain (T4IRD and T3IRD) are poised toward T4 and T3 degradation, while that in liver (T4ORD) is poised toward T3 generation. PMID- 8729943 TI - Effect of dominance status on sex hormone levels in laboratory and wild-spawning male trout. AB - We investigated the relationship between male social status and hormone levels in salmonids spawning under laboratory and field conditions. In small groups of rainbow trout (Onchorhynchus mykiss) spawning in the laboratory, dominant males had higher plasma levels of testosterone (T) and 17 alpha, 20 beta-dihydroxy-4 pregnen-3-one (17,20 beta-P) compared with subordinates. Steroid levels increased in subordinate males that became dominant after dominant males were experimentally removed; higher steroid levels in dominant males appears to be a result rather than a cause of their social status. In free-ranging brown trout (Salmo trutta) sampled in the field, we found higher levels of 11 ketotestosterone (11KT) but not T in dominant males. No significant differences in levels of either androgen were found between dominant and subordinate male brook trout (Salvelinus fontinalis) sampled at the same field location. Furthermore, in marked contrast with the laboratory fish, there were no significant differences in plasma 17,20 beta-P between dominant and subordinate males in either species of fish in the wild. The different findings in the laboratory and field may indicate species differences in behavioral endocrinology among brook, brown, and rainbow trout. Alternatively, the greater differential in hormonal profile of dominant and subordinate males in the laboratory may reflect the relative uniformity of the laboratory environment; this simple environment may allow competitively superior males to more completely dominate less competitive tank-mates and to exclude them from female sexual cues. In any case, these results suggest that the relationship between steroid hormones and spawning behavior in male salmonids is likely more complex than suggested by experiments conducted solely on laboratory-held rainbow trout. PMID- 8729944 TI - Developing gossypol derivatives with enhanced antitumor activity. AB - Preclinical and clinical studies have pointed to the antitumor potential of the naturally occurring polyphenolic binaphthyl dialdehyde, gossypol, as well as its purified (-,+) enantiomers. To explore further the antitumor properties of this multifunctional agent, we synthesized several reactive derivatives including the (-,+) enantiomers of gossypolone and four different gossypol Schiff's bases (AR1, AR2, AR3, AR4). The biological activities of these new agents were screened by measuring their in vitro antiproliferative activity against malignant (MCF-7, MCF 7/adr) or immortalized (HBL-100) human breast epithelial cell lines. Racemic gossypolone showed relatively uniform antiproliferative activity against all of the breast epithelial cell lines with 3- to 5-fold less activity than (--) gossypol against MCF-7 and MCF-7/adr cells. Of interest, the relative antitumor potency of purified gossypolone enantiomers was reverse that of gossypol enantiomers, since (+)-gossypolone showed up to 3-fold greater inhibition of MCF 7 culture growth than (--)-gossypolone. Of the Schiff's base derivatives only AR3 with its isopropyl amine substituent demonstrated cytotoxic activity comparable to that of (--)-gossypol; derivatives with ethyl, propyl, or butyl amine substituents (AR1, AR2, AR4) had little growth inhibitory activity at culture concentrations up to 25 microM. AR3 activity was greatest against HBL-100 and MCF 7 cells [MCF-7 IC50 values: AR3 = 0.9 microM, (--)-gossypol = 2.3 microM]; unlike (--)-gossypol, however, AR3 showed substantially reduced activity against the multidrug-resistant subline, MCF-7/adr. These structure-activity comparisons suggest that isolation of (-,+)-enantiomers of AR3 and additional chemical modifications including the synthesis of an isopropyl amine Schiff's base of gossypolone will likely yield a newer generation of gossypol analogues with enhanced anticancer potential. PMID- 8729945 TI - Effects of vinorelbine on epidermal growth factor-receptor binding of human breast cancer cell lines in vitro. AB - Epidermal Growth Factor (EGF) is a mitogenic peptide that binds to surface membrane receptors (EGFR) of breast cancer cells. After binding, secondary transmitter molecules are activated by tyrosine phosphorylation of the intracellular receptor domaine. The activity of the EGF/EGFR system can be modulated by a variety of chemically unrelated compounds including cytostatic agents. The purpose of our present study was to determine the effects of vinorelbine, a novel semisynthetic vinca alkaloid on EGF receptor binding on human breast cancer cells. We have found that MDA-231 and MDA-468 cells bind substantially more [125I]-EGF after preincubation with vinorelbine. This effect was concentration- and time-dependent reaching a maximum at 100 ng/ml and 24 h incubation. Subsequent experiments showed an increase in the rate of EGF binding as well as maximal binding capacity. Scatchard analysis of binding experiments under equilibrium conditions indicated that this was mainly due to an increase in the number of apparent EGF binding sites. Modulation of EGF receptor binding by vinorelbine was not detectable when isolated membranes were used indicating that intact cytoplasmatic mechanisms are required for the upregulation of EGF receptors. PMID- 8729946 TI - Comparative efficacy of DMP 840 against mouse and human solid tumor models. AB - BACKGROUND: DMP 840 is a compound from a class of bis-naphthalimide antitumor agents that recently completed Phase I clinical trials at three North American centers and is currently undergoing Phase II testing. Preclinically, it was shown to have curative activity against a variety of human tumor xenograft models. PURPOSE: To test DMP 840 both in vitro and in vivo for antiproliferative activity against predominantly mouse tumor models. METHODS: A disk diffusion soft agar colony formation assay was used to determine the in vitro growth inhibitory activity against a selection of mouse and human tumor cell lines, and the comparable selective mouse solid tumors were used for in vivo testing. RESULT: In vitro DMP 840 exhibited equal cytotoxicity for human tumors (including MX-1 directly cultured from nude mice), mouse tumors and normal cells. In vivo DMP 840 was only modestly active or inactive against the following mouse tumors: Mam 16/C, T/C = 30% (T/C = Percent Tumor Growth Inhibition); Mam 16/C/ADR, T/C = 33%; Colon 38, T/C = 9%; Panc 03, T/C = 53%; Colon 51/A, T/C = 28%; Panc 02, T/C = 52%; P388/0, 36% ILS (Percent Increased Life Span) and P388/ADR, 14% ILS. Furthermore, the antitumor activity was only observed at the highest non-toxic dose and was associated with a large body weight loss. In contrast, the agent was highly active against the human breast tumor MX-1 implanted subcutaneously in either athymic nude or SCID mice (Nudes: T/C = 0%; 1/5 cures; SCIDS: T/C = 0%; 5/5 cures). CONCLUSIONS: Although there was no selective cytotoxicity in our clonogenic assay for human versus mouse tumor cell lines, selective activity in vivo for human xenograft tumors was noted. Overall, this compound is rather unique in its differential degree of in vivo activity for human versus mouse tumors. IMPLICATIONS: Phase II trials, which are ongoing, will help determine if the preclinical in vivo selective activity of DMP 840 translates to clinical activity in man. PMID- 8729947 TI - Phase I trial of ilmofosine as a 24 hour infusion weekly. AB - Ilmofosine, an ether lipid derivative of lysophosphatidylcholine has antineoplastic activity in vitro and in vivo. Maximum efficacy in preclinical models is associated with prolonged exposure to the drug. In a Phase I trial of a weekly 2 hour infusion schedule of ilmofosine, a syndrome of lethargy, diminished performance status, and mild hepatotoxicity was dose-limiting at 550 mg/m2. To avoid the higher drug concentrations associated with a brief infusion, a Phase I study of a weekly 24 hour infusional schedule was undertaken in an attempt to maximize dose-intensity. Doses were escalated from 550 to 800 mg/m2. Toxicities included nausea, anorexia, fatigue, and minor elevations of liver function tests. The dose limiting toxicity at 800 mg/m2 was a syndrome of severe abdominal pain. No neutropenia or thrombocytopenia was observed except in one patient who was found to have a myelodysplastic syndrome, thought not to be related to drug therapy. The more prolonged infusion schedule of ilmofosine did not result in a substantial increase in the tolerable dose. PMID- 8729948 TI - A phase I study of acivicin in refractory pediatric solid tumors. A Pediatric Oncology Group study. AB - Forty-two patients with progressive solid tumors and brain tumors were entered in this Phase I study of the glutamine antagonist acivicin given intravenously over thirty minutes daily for five days. The major toxicities encountered were myelosuppression and central nervous system toxicity (nightmares and somnolence). The maximum tolerated dosage on this schedule was 26 mg/M2 daily for five days. Six patients including three patients with brain tumor had stable disease. PMID- 8729949 TI - A phase II trial of paclitaxel (Taxol) as first line treatment in advanced breast cancer. AB - Paclitaxel (Taxol) is a natural product with a broad spectrum of activity against various solid tumors. This report includes nineteen patients with advanced breast cancer who have not previously received chemotherapy for metastatic disease. Fifteen patients had received adjuvant chemotherapy, eight of which were doxorubicin based. Patients were treated with 135 mg/m2 over 24 hours by continuous infusion given every 21 days. There were 2 complete and 4 partial responses for an objective response rate of 32% (95% C.I.: 14%, 57%) and eight patients or 42% with stable disease. Three of eight patients (38%) who had received adjuvant doxorubicin did respond to paclitaxel. Responses occurred in lung, liver, and soft tissue. The primary toxicity was hematologic with 13 hospitalizations for febrile neutropenia in 180 cycles (7%). Paclitaxel has moderate activity in a small number of patients with metastatic breast cancer at the dose of 135 mg/m2 over 24 hours in this study. PMID- 8729950 TI - Phase II trial of taxol in patients with adenocarcinoma of the upper gastrointestinal tract (UGIT). The Eastern Cooperative Oncology group (ECOG) results. AB - Taxol was administered as a 24-hour continuous infusion at 250 mg/m2 in this Phase II trial in patients with adenocarcinomas of the upper gastrointestinal tract (UGIT). Twenty-five patients were entered between July 1991 and June 1992, twenty-three were eligible and were evaluated for toxicity and twenty-two were assessable for response. There was one partial response (4.5%) in a patient with liver metastases, with a duration of 6 months. Toxicity was primarily neutropenia. Taxol as a single agent appears to have little activity in adenocarcinoma of the UGIT. PMID- 8729952 TI - Phase II study of topotecan in metastatic hormone-refractory prostate cancer. AB - Systemic chemotherapy with currently available agents has not improved survival for patients with hormone refractory prostate cancer (HRPC), consequently, the evaluation of new agents is warranted. Topotecan is a specific inhibitor of topoisomerase I with broad antitumor activity in preclinical studies. The purpose of this phase II trial was to determine the objective response rate of topotecan administered as a 30 minute infusion for five consecutive days in men with metastatic HRPC. Thirty-four evaluable patients were treated with topotecan 1.1 1.5 mg/m2 as a 30 minute infusion daily for five days, repeated every three weeks until disease progression or unacceptable toxicity. Response was assessed with a combination of standard solid tumor response criteria and the serum prostate specific antigen (PSA) for patients with bidimensionally measurable disease, and by serial measurements of the PSA in patients with bone only (evaluable) disease. One of 13 patients (7.6%) with measurable soft tissue disease had a PR in nodal sites. Of 21 patients with only osseous metastases, 1 (4.7%) had improvement in bone scan. Six of the 34 evaluable patients (17.6%) had the serum PSA decrease by > or = 50% and 2 (5.8%) had PSA decreases of > or = 75%. Toxicity was chiefly hematologic with 66% of patients experiencing Grade 3 or 4 granulocytopenia. Thirty-nine percent of cycles required a delay to allow for hematologic recovery and ten patients required red cell transfusions. Non-hematologic toxicity, mainly nausea and alopecia, was mild. Topotecan administered at this dose and schedule has limited activity in patients with HRPC. Further trials of topo I inhibition in HRPC should utilize alternative schedules of topotecan (e.g., prolonged infusion) or other camptothecin analogs with more potent topo I inhibitory activity. PMID- 8729951 TI - A phase II trial of weekly infusional 5-fluorouracil in combination with low-dose leucovorin in patients with advanced colorectal cancer. AB - Exogenous leucovorin is a source of reduced folate which enhances the inhibition of thymidylate synthase that results from 5-fluorouracil (5-FU) administration. Extracellular reduced folate concentrations of 1 microM have been reported to yield maximal enzyme inhibition in several cell lines treated with 5-FU in vitro. Clinical studies indicate that low doses of leucovorin have equivalent efficacy to higher doses in successfully modulating 5-FU in the treatment of colorectal cancer. Based on pharmacokinetics at higher doses, steady-state total plasma reduced folate concentrations of 1 microM would be expected from the administration of leucovorin 50 mg/m2 by 24 h infusion. This dose was admixed with 5-FU 2300 mg/m2 and administered by 24 h-infusion weekly to 38 patients with advanced colorectal cancer, of whom 32 are evaluable for response. Disease sites included liver (33 patients), lung (12 patients), and bone (4 patients). Toxicity was mild to moderate, except for grade 3 diarrhea in 5 patients, and chest pain in 2 patients. Among the 32 evaluable patients, there were 14 partial remissions for a total response rate of 44% (95% confidence interval 27-61%). The median duration of response was seven (range 1 to 20+) months, and median duration of survival 16 months. These results support the use of low doses of leucovorin to modulate weekly infusional 5-FU in colorectal cancer, and provide a basis for the integration of this regimen with other modulators of 5-FU. PMID- 8729953 TI - A phase II study of interleukin-2 with and without beta-interferon in the treatment of advanced renal cell carcinoma. AB - BACKGROUND: Biologic response modifiers have activity in renal cell carcinoma. The combination of interleukin-2 (IL-2) and beta-interferon (B-IFN) is synergistic in vitro. This trial was initiated to determine the efficacy of IL-2 alone and with B-IFN in advanced RCC. METHODS: Ambulatory patients with advanced RCC were randomly allocated to either IL-2 6 x 10(6) units/M2 intravenously (IV) three days a week for four weeks or IL-2 5 x 10(6) units/M2 IV plus B-IFN 6 x 10(6) units/M2 IV three days a week for 4 weeks. This induction phase was followed by a maintenance phase of the same drugs and doses administered for two weeks out of every four. RESULTS: 84 patients were entered onto this phase II trial with 75 considered eligible for response and survival. Toxicity is reported for the 81 patients on whom data was received, irrespective of eligibility. The overall response rate (RR) was 9.3% (7/75). Of the 3 responses in the IL-2 arm (RR = 8.3%), one was a complete response. 4 patients in the IL-2 + B-IFN arm (RR = 10.3%) achieved a partial response. Median survival was estimated to be 8.4 months for patients given IL-2 and 8.0 months for patients given the IL-2 and B IFN combination. Multivariate analysis of survival data identified initial performance status, metastases of > 1 site, and weight loss as being important prognostic factors for survival. There were 2 lethal and 3 life threatening toxicities with the IL-2 treatment. While there were no lethal toxicities on the combination arm, there were 4 life threatening toxicities. CONCLUSIONS: The results of this study indicate that further investigation of IL-2 with or without B-IFN at this dose and schedule as treatments for renal cell carcinoma is probably not warranted. PMID- 8729954 TI - Phase II study of amonafide in the treatment of patients with advanced squamous cell carcinoma of the head and the neck. An Illinois Cancer Center study. AB - Amonafide (nafidimide), a synthetic organic compound with an inhibitory effect on cellular replication, was used in a phase II study conducted by the Illinois Cancer Center in order to assess its efficacy and toxicity in advanced or recurrent squamous cell cancer of the head and neck. Eligible patients had received no more than one prior adjuvant or neoadjuvant chemotherapy, had normal bone marrow, renal and hepatic function, ECOG performance status of 0-2, and bidimensionally measurable disease. Eligible patients were administered amonafide at a starting dose of 300 mg/m2 for five consecutive days every 3 weeks with dose escalation or de-escalation according to established hematologic criteria in the absence of disease progression. Nineteen of 22 entered patients were evaluable for response and all patients were evaluable for toxicity. Eleven of 19 patients achieved stable disease. Median time to progression after start of treatment was 57 days, for the 18 patients for whom the date of progression is known. There were no partial or complete responses. Hematologic toxicity was dose limiting with grade 3-4 neutropenia in 50 percent of patients and 4 deaths associated with neutropenic sepsis. Non-hematologic toxicity was mild to moderate with nausea and vomiting predominating. In this study, amonafide was a myelotoxic, inactive treatment in advanced/recurrent head and neck cancer. Further use in head and neck cancer appears unwarranted. PMID- 8729955 TI - A clinical screening cooperative group phase II evaluation of lobaplatin (ASTA D 19466) in advanced head and neck cancer. AB - We evaluated the efficacy and tolerability of lobaplatin, a new platinum compound, given at the dose of 50 mg/m2 by i.v. bolus every 4 weeks, in 49 patients with advanced and/or metastatic squamous cell carcinoma of the head and neck (SCCHN). One complete and 2 partial responses were observed in 43 eligible patients for an overall response rate of 7% (95% confidence interval: 1-19%). The duration of responses was 11, 16 and 32 weeks. Toxicities of WHO grade > or = 3 were hematologic: thrombocytopenia in 26%, granulocytopenia in 12% and anemia in 12% of patients. There was no therapy-related death. Nausea/vomiting, diarrhoea and paresthesia were mild and rare. In conclusion, lobaplatin was well tolerated, but its efficacy in advanced SCCHN at the presented dose and schedule, was marginal. PMID- 8729956 TI - Didemnin B in favourable histology non-Hodgkin's lymphoma. A phase II study of the National Cancer Institute of Canada Clinical Trials Group. AB - Ten patients with previously untreated stage III/IV low grade histology non Hodgkin's lymphoma received a 1-hour intravenous infusion of Didemnin B 2.3 mg/m2 weekly for 4 weeks repeated every 6 weeks. 40% of patients experienced significant hypersensitivity reactions, one of which was life-threatening, despite premedication with diphenhydramine and cimetidine. Other toxicities included nausea, vomiting, fatigue, diarrhea and skin rashes. No objective responses were seen. Given the serious toxicity and lack of activity in a non pretreated group of patients, the study was closed early. Further investigation of Didemnin B at this dose and schedule is not recommended. PMID- 8729957 TI - Cutaneous manifestations of Taxol therapy. AB - Taxol is a novel chemotherapeutic agent that has produced substantial responses in early clinical studies [1]. Taxol has excellent activity in a number of malignancies based on recently completed clinical trials, including a 30% response rate in platinum-refractory ovarian cancer patients [2-5]. We are currently conducting trials of dose-intense taxol with granulocyte colony stimulating factor (G-CSF) support in relapsed or refractory ovarian cancer patients. Such dose intensification produces a major response rate in 50% of patients with this disease [6]. Taxol was supplied in 5 ml ampules (6 mg/ml) in polyethoxylated castor oil (Cremophor EL) 50% and dehydrated alcohol and the dose was diluted in either 0.9% sodium chloride or 5% dextrose at concentrations of 0.6 to 1.2 mg/ml. We have noted 3 patients with previously unreported cutaneous manifestations which we believe are taxol related and also report our overall complication rate with the administration of taxol by peripheral intravenous lines. PMID- 8729958 TI - Molecular aspects of pesticide degradation by microorganisms. AB - Microorganisms are able to degrade a large variety of compounds, including pesticides under laboratory conditions. However, methods have yet to be developed to decontaminate the environment from residues of pesticides. Pesticidal degradative genes in microbes have been found to be located on plasmids, transposons, and/or on chromosomes. Recent studies have provided clues to the evolution of degradative pathways and the organization of catabolic genes, thus making it much easier to develop genetically engineered microbes for the purpose of decontamination. Genetic manipulation offers a way of engineering microorganisms to deal with a pollutant, including pesticides that may be present in the contaminated sites. The simplest approach is to extend the degradative capabilities of existing metabolic pathways within an organism either by introducing additional enzymes from other organisms or by modifying the specificity of the catabolic genes already present. Continuous efforts are required in this direction, and at present several bacteria capable of degrading pesticides have been isolated from the natural environment. Catabolic genes responsible for the degradation of several xenobiotics, including pesticides, have been identified, isolated, and cloned into various other organisms such as Streptomyces, algae, fungi, etc. In addition, recombinant DNA studies have made it possible to develop DNA probes that are being used to identify microbes from diverse environmental communities with an unique ability to degrade pesticides. PMID- 8729959 TI - Ecology, metabolism, and genetics of ruminal selenomonads. AB - Selenomonas ruminantium is one of the more prominent and functionally diverse bacteria present in the rumen and can survive under a wide range of nutritional fluctuations. Selenomonas is not a degrader of complex polysaccharides associated with dietary plant cell wall components, but is important in the utilization of soluble carbohydrates released from initial hydrolysis of these polymers by other ruminal bacteria. Selenomonads have multiple carbon flow routes for carbohydrate catabolism and ATP generation, and subspecies differ in their ability to use lactate. Some soluble carbohydrates (glucose, sucrose) appear to be transported via the phosphoenolpyruvate phosphotransferase system, while arabinose and xylose are transported by proton symport. High cell yields and the presence of electron transport components in Selenomonas strains has been documented repeatedly and this may partially account for the energy partitioning observed between energy consumed for growth and maintenance functions. Most strains can utilize ammonia, protein, and/or amino acids as a nitrogen source. Some strains can hydrolyze urea and/or reduce nitrate and use the ammonia for the biosynthesis of amino acids. Experimental evidence suggests that ammonia assimilatory enzymes in some strains may possess unique properties with respect to other presumably similar bacteria. Little is known about the genetics of ruminal selenomonads. Plasmid DNA has been isolated from some strains, but it is unknown what physiological functions may be encoded on these extrachromosomal elements. Due to the predominance of S. ruminantium in the rumen, it is an ideal candidate for genetic manipulation. Once the genetics of this bacterium are better understood, it may be possible to amplify its role in the rumen. PMID- 8729960 TI - Spatial and temporal distribution of lamprin mRNA during chondrogenesis of trabecular cartilage in the sea lamprey. AB - The temporal and spatial expression patterns of lamprin, the principle structural protein in lamprey cartilages, were examined by in situ hybridization during chondrogenesis of trabecular cartilage in day 17-33 post-fertilization prolarval lampreys. Lamprin mRNA transcripts were first detected during day 19, concomitant with the end of the condensation phase of chondrogenesis and the initiation of matrix synthesis as indicated by light microscopic examination. In the stages which followed, the hybridization signal increased with progressive intensity, paralleling matrix synthesis, suggesting transcriptional control of lamprin gene expression. Spatially, lamprin expression patterns mirrored the rostrocaudal development of the trabecular cartilage rudiment. No signal was detected over adjacent tissues or control sections. Some similarities exist between the temporal patterns of lamprin expression and the expression of matrix proteins such as elastin and collagen of higher vertebrates. It is concluded that certain aspects of chondrogenesis are critical to the normal development of a functional cartilaginous matrix and are conserved throughout the vertebrate taxa. PMID- 8729961 TI - Comparative distribution of NADPH-diaphorase activity and tyrosine hydroxylase immunoreactivity in the diencephalon and mesencephalon of the domestic chicken (Gallus domesticus). AB - We described the distribution of NADPH-diaphorase-containing neurons in relation to tyrosine hydroxylase immunoreactivity in the diencephalon and mesencephalon of the chicken. In the diencephalon, both markers were found in the lateral hypothalamus, dorsal hypothalamic area, hypothalamic periventricular nucleus, paraventricular nucleus and mamillary area. A close examination showed that the fine distribution of these markers differed slightly, so that they were never observed in the same neurons. In the mesencephalon, NADPH-diaphorase and tyrosine hydroxylase immunoreactivity were found in the ventral pedunculopontine area (nucleus tegmenti pedunculopontinus pars compacta, adjacent areas surrounding the quintofrontal tract and the nucleus mesencephalicus profundus ventralis), the coeruleus complex (locus coeruleus, ventral and dorsal subcoeruleus nuclei), the ventral tegmental area and the central gray. The majority of these neurons contained either diaphorase or tyrosine hydroxylase. Nevertheless, in a few cases both markers appeared to colocalize in the same neuron, typically in large perikarya of the ventral pedunculopontine area. PMID- 8729962 TI - Early organization of the pituitary gland in Sparus aurata L. (Teleostei). An ultrastructural study. AB - The cell organization of the pituitary gland and the relationship between neurohypophysis and adenohypophysis in early developmental stages of the gilthead sea bream, Sparus aurata, were studied by electron microscopy. In newly hatched larvae, the pituitary gland was embedded in the ventral floor of the diencephalon and separated from the hypothalamus by a continuous basal lamina. Elongated mesenchymal cells next to the ventral surface were observed. At this stage, there was no neurohypophysis and the adenohypophysis consisted of undifferentiated endocrine cells with small scarce secretory granules and a few stellate cells, with no distinctive zonation. An incipient neurohypophysis was present in 1-day old larvae. The first evagination of the neurohypophysis into the adenohypophysis were observed in 2-day-old larvae and developed progressively with age, being deeper in the caudal zone. Two regions in the adenohypophysis, one anterior--the presumptive pars distalis--and one posterior--the presumptive pars intermedia- were found in 2-day-old larvae. Three regions (rostral and proximal pars distalis and pars intermedia) were clearly distinguishable in 4-day-old larvae. The ultrastructural features of the pituitary endocrine cells varied during gland differentiation, with the secretory granules gradually increasing in number and size, accompanying organelle development. Nevertheless, even in the oldest larvae studied (65 days), undifferentiated cells similar to those in the earliest stages were observed. The first blood vessels appeared in the neurohypophysis around 16 days after hatching. During early development, the pituitary gland progressively emerged from the ventral floor of the brain. By 16 days, the principal pattern of the pituitary gland architecture appeared to be established. PMID- 8729963 TI - Characterization of chondrogenesis in cells isolated from limb buds in mouse. AB - Micromass cultures of mesenchymal cells isolated from limb buds of 11.5-day-old mouse fetuses were used to study chondrogenesis. After 3 days of culture, dense cell aggregates were observed. They then were converted into macroscopically visible cartilage foci during the following 2-4 days. Comparison of 2-, 4- and 7 day-old cultures has shown that the cells first expressed collagen type I, then switched to collagen type II expression as shown by immunohistochemistry and in situ hybridization. At day 7, proteoglycans were synthesized centrally in the foci. At the same time, most cells expressed collagen type II, with the highest expression in the periphery of the aggregates. The oncogene c-fos and homeodomain protein FS-1 were found in the cells expressing collagen type II, indicating that these transcription factors may be involved in the regulation of cell differentiation. The expression of alkaline phosphatase was detected first in mature cartilage foci (day 4) and increased during culture. Early in culture, DNA replicating cells were uniformly distributed. With differentiation, the proliferating cells were present predominantly between the aggregates and their total number became significantly reduced. Our results indicate that the process of chondrogenesis in micromass cultures of mesenchymal cells mimics the differentiation process occurring during fetal development in vivo and can be directly studied by in situ hybridization, immunohistochemical and histochemical methods. PMID- 8729965 TI - Dorsal root ganglion neuron development in chick and rat. AB - The aim of this study was to compare the morphological development of dorsal root ganglion neurons in embryonic and early postnatal chicks and rats. The three dimensional architecture of neurons was observed in ganglia in situ and in dissociated neurons by scanning electron microscopy after removal of the capsule and connective tissue. The percentages of neurons at different developmental stages were determined. The general morphological changes in the chick resembled those in the rat but the timing was different. In both chick and rat, the majority of neurons were bipolar at early stages of development (embryonic day 6 in chick and day 14 in rats) and later underwent pseudo-unipolarization to become mostly unipolar neurons at hatching or birth. This maturation event started at an earlier stage in chick embryos than in rats, with 57% unipolar neurons in chick and only 7% in rat on embryonic day 14. However, just after hatching or birth, at day 22 of development, a larger proportion of immature unipolar neurons remained in chicks (13%) than in rats (3%). We conclude that these differences should be taken into consideration in designing experiments on dorsal root ganglion neurons grown in tissue culture. PMID- 8729964 TI - The raccoon lateral cervical nucleus: mediolateral organization of GABA-positive and GABA-negative neurons and fibers. AB - In the lateral cervical nucleus (LCN) of the cat, GABA-immunoreactive neurons and substance P-immunoreactive fibers are concentrated in the medial part of the nucleus, whereas in the monkey LCN no preferential locations have been identified. In raccoons, substance P-immunoreactive fibers display a distribution pattern similar to that in cats. However, the presence and distribution of GABA immunoreactive neurons in the raccoon LCN has not been examined, and it is therefore not known whether raccoons are similar to cats or primates in this respect. Thus, in the present study, the raccoon LCN was examined for the presence and distribution of GABA-immunoreactive cells with respect to their numbers, locations, and sizes. The distribution of GABA-positive fibers and varicosities within the LCN was also investigated. The results of measurements of cross-sectional areas of LCN neurons indicate a trend toward decreasing cell size along the dorsolateral to medial axis of the raccoon LCN. Compared to neurons of the centrally located ventromedial division, neurons are statistically significantly larger in the dorsolateral division and smaller in the medial division of the nucleus. Cell counts in post-embedding-stained semithin sections through the nucleus revealed an average of 8,700 neurons per LCN. Approximately 4% of LCN neurons are GABA-immunoreactive. These neurons are small and most (80%) of them are located in the medial third of the LCN. In contrast, GABA immunoreactive fibers and varicosities are present in about equal density throughout the raccoon LCN. Thus, the distributions of GABA-immunoreactive neurons and neuron sizes in the raccoon LCN conform closely to those in cats. Together with previous observations in cats and raccoons, the present findings support the notion that these small GABA-immunoreactive neurons may be local circuit inhibitory neurons and indicate the presence of a mediolateral segregation that may be of fundamental importance for the functional organization of the carnivore LCN. PMID- 8729966 TI - Organization of identified fiber tracts in the rat fimbria-fornix: an anterograde tracing and electron microscopic study. AB - The fimbria is a major route for afferent and efferent fibers of the hippocampal formation. However, little is known about the intrinsic organization of the fimbria-fornix complex. In this study, the anterograde tracer Phaseolus vulgaris leucoagglutinin (PHAL) was used to analyze the ultrastructure and topography of identified fiber tracts within the fimbria-fornix. Septo-hippocampal fibers are loosely distributed throughout the fimbria-fornix. Commissural fibers cross the midline in the ventral hippocampal commissure and form a tight fiber bundle in the fimbria. Crossed entorhino-hippocampal fibers cross the midline in the ventral hippocampal commissure rostral to the commissural fiber bundle, and crossed entorhino-entorhinal fibers pass through the dorsal hippocampal commissure. This suggests a topographical organization of fiber tracts within the fimbria-fornix that reflects the laminar organization of the hippocampal target structure: fibers of the diffusely terminating septohippocampal projection are loosely distributed throughout the fimbria-fornix, while those projections that are known to terminate in specific laminae of the hippocampal formation (commissural projection, crossed entorhino-hippocampal projection) form fiber bundles within the fimbria and the ventral hippocampal commissure. PMID- 8729967 TI - Cellular cementogenesis in rat molars: the role of cementoblasts in the deposition of intrinsic matrix fibers of cementum proper. AB - The formation of intrinsic fibers was examined in the advanced stage of rat cellular cementogenesis by light and electron microscopy. Using scanning electron microscopy, cementoblasts showed wing-like processes, partly encircling principal fibers. At the cementum-facing side of the cells these processes showed segmentation into finger-like processes, arranged in parallel with the cementum surface. Transmission electron microscopy showed many cytoplasmic fragments around intrinsic fibers at the cementum surface. These fragments contained microtubules and collagenous secretory granules that were arranged in parallel with the cementum surface and the intrinsic fibers. The wing-like processes contained microtubules and secretory granules that were arranged perpendicularly to the cementum surface and in parallel with the principal fibers. These observations suggest that: (1) the cytoplasmic fragments are cross-sectioned finger-like processes; (2) cementoblasts secrete intrinsic fibers from the finger like processes and additional principal fibers from the wing-like processes; (3) cementoblasts constantly shorten their wing-like processes by forming finger-like processes. This development starts at the side facing the cementum and proceeds towards the periodontal ligament. With the segmentation, the cementoblasts change the arrangement of secretory granules to secrete intrinsic fibers around preformed principal fibers. PMID- 8729968 TI - Development of cardiac musculature in the cranial vena cava of rat embryos. AB - Development of cardiac musculature in the rat cranial vena cava (common cardinal vein or duct of Cuvier) was examined by immunohistochemistry and transmission electron microscopy. Undifferentiated cardiac myocytes were detected in the cranial vena cava wall of rat embryos after 12.5 days post-coitum (dpc). The tunica media of the cranial vena cava was composed of cardiac myocytes after formation of the endothelium. Therefore, the cranial vena cava may be not only a part of the venous system but also of the heart. Myocytes in the cranial vena cava contained developing myofibrils, mitochondria and intercalated discs similar to those found in the myocytes in heart. Striated myofibrils began to differentiate as soon as myocytes appeared in the vena cava wall, and myocytes with differentiating myofibrils occur in the wall as the first component of the tunica media at 12.5 dpc. We concluded that the cardiac musculature in the vena cava is not a secondary extension into the tunica media after birth only in the rat, but a basic structure formed in all mammals during early embryonic development. PMID- 8729969 TI - Chondroid tissue in the early facial morphogenesis of the chick embryo. AB - The calcified tissues involved in the early morphogenesis of the so-called intramembranous bones of the facial skeleton were studied by microradiographic and histological techniques in 22 chick embryos at the 9th, 12th and 14th days of incubation. On the 9th day, the bones of the upper face and palatal vault are made up of thin sheets of chondroid tissue, deposited in their respective mesenchymal condensations. Woven and lamellar bone formation subsequently takes place in each of them from the 12th day of incubation, mainly on the external side of their chondroid primordia. The same phenomena occur in the lower facial and mandibular bones. These facts indicate that the primitive facial desmocranium of the chick embryo, which is classically considered to be formed by intramembranous ossification, first consists of chondroid tissue. As in the cranial vault, this tissue thus represents the initial modality of the skeletogenic differentiation within the avian facial mesenchyme. PMID- 8729970 TI - Is the meaning of science changing in the practice of medicine? PMID- 8729971 TI - mdm-2 oncoprotein expression associated with deletion of the long arm of chromosome 12 in a case of mantle cell lymphoma with blastoid transformation [corrected]. AB - We report a unique case of mantle cell lymphoma in blastoid transformation associated with deletion of the long arm of chromosome 12 and with 90 kDa mdm-2 protein overexpression. Neither the mantle cells nor their blastoid counterparts expressed p53 gene product by immunohistochemical analysis. This seems to be the first reported case of this subtype of lymphoma associated with these specific cytogenetic and molecular genetic abnormalities. PMID- 8729972 TI - Molecular diagnosis of mantle cell lymphoma in paraffin-embedded tissue. AB - Mantle cell lymphoma (formerly known as intermediate lymphocytic lymphoma, diffuse centrocytic lymphoma, or diffuse small cleaved lymphoma) is one of the small cell non-Hodgkin lymphoma entities that is clinically more aggressive than small lymphocytic lymphoma, and needs to be separated from it. Mantle cell lymphoma is strongly associated with the t(11;14) chromosomal translocation that rearranges the bcl-1 oncogene (PRAD-1 gene) and immunoglobulin heavy chain gene. In this study, we developed a nested polymerase chain reaction system to evaluate the t(11;14) translocation. The study material consisted of 10 mantle cell lymphomas fulfilling the criteria suggested by other authors (P. M. Banks et al. Surg Pathol 16:637, 1992). A novel nested polymerase chain reaction system was used to evaluate the bcl-1 breaks in the major translocation cluster using two successive polymerase chain reaction amplifications. This reaction yielded a background-free single product of the size of 200 to 300 base pairs in four of 10 mantle cell lymphomas. The identity of the product from the nested polymerase chain reaction was confirmed by Southern blotting followed by hybridization with a specific probe. The amplification products were also evaluated by Sst-I, Alu-I, Dde-I, and Ita-I restriction enzymes and showed different patterns of digestion reflecting individual differences between the MTC/ IgH junctions. A selection of other low-grade lymphomas, including lymphocytic, follicular, and mucosa associated lymphoid tissue lymphoma, and hairy cell leukemia and 29 hyperplastic lymph nodes were negative. This nested polymerase chain reaction system for the t(11;14) translocation involving major translocation cluster offers a convenient specific identification for mantle cell lymphoma. However, this test has a limited diagnostic power because only about half of the mantle cell lymphomas show the bcl-1 breaks in the major translocation cluster. The test performs well in formaldehyde-fixed and paraffin-embedded material, allowing the study of large numbers of retrospective cases of mantle cell lymphomas. PMID- 8729973 TI - ras and p53 genes in male breast cancer. AB - We have studied the roles of Ki-ras oncogene and p53 tumor suppressor gene in a series of 20 cases of male breast cancer and one papilloma of the male breast. Ki ras was detected in 50-microns sections after digestion with proteinase K and SDS. DNA was amplified by polymerase chain reaction, dot blotted, and mutations were screened with labeled ras mutation-specific oligonucleotides. Wild-type and mutant p53 protein were detected with antibodies CM1 and DO7, using the avidin biotin-peroxidase method. Two of 17 carcinomas showed Ki-ras mutations, both in codon 12 (gly --> lys and gly --> arg). Five of 20 male breast cancers (25%), including one large intraductal carcinoma, expressed mutant p53 protein. Although the incidence of mutant p53 expression in male breast cancer is similar to that in women, Ki-ras mutations are not significantly increased. PMID- 8729974 TI - Mixed testicular germ cell tumors: monoclonal or polyclonal. AB - Testicular germ cell tumors (TGCT) of adolescents and adults are, for clinical and pathological reasons, divided in seminomas (SE) and nonseminomatous germ cell tumors (NS). Whether and to what degree these two entities are pathogenetically related is still controversial and a matter of debate. TGCT may contain both SE and NS components. Cytogenetic studies of the SE and NS components of these mixed TGCT might shed light on the pathogenetic relationship between both components. Separate cytogenetic analysis was performed on both components of three cases of mixed TGCT with both SE and NS components. The karyotypes of both components were compared with each other. In one case, the SE and NS component share eight different structural chromosomal abnormalities, indicating that the SE and NS component are pathogenetically closely related and have a common neoplastic pathway for a considerable length. Both components of the other two cases have, respectively, i(12p) and no structural chromosomal abnormalities in common. Our results, together with data from the literature, indicate that in mixed TGCT with SE and NS components, both components may have either a monoclonal or a polyclonal origin. PMID- 8729976 TI - Abdominal wall implantation of an endocervical-like mucinous borderline tumor. AB - We present the case of a 21-year-old woman in whom development of a bulky abdominal wall metastasis in a laparotomy scar led to discovery of bilateral endocervical-like mucinous borderline tumors in normal-sized ovaries. Tumor implants were also present in the mesosalpinx and a pelvic lymph node. We hypothesize that the abdominal wall metastasis resulted from seeding at the time of a prior exploratory laparotomy for trauma, before the ovarian tumors were discovered. We present evidence to support our theory of mechanical implantation of borderline tumor and explore other mechanisms leading to extraovarian mucinous neoplastic involvement. PMID- 8729975 TI - Pseudobowenoid change of the vulva: a histologic variant of untreated condylata acuminatum. AB - Vulvar condylomata are distinguished from vulvar intraepithelial neoplasia by orderly maturation, a low mitotic index, and the absence of nuclear atypia in the lower epithelial layers. Podophyllin therapy might produce changes in condylomata mimicking vulvar intraepithelial neoplasia but can be distinguished, particularly with a recent history of podophyllin use. This report describes five patients with vulvar condylomata that morphologically resemble both podophyllin effect and vulvar intraepithelial neoplasia. The mean age was 30.5 years. All lesions exhibited superficial keratinocytes in various stages of apoptosis, with a spectrum of nuclear degenerative changes characterized by absence of the nuclear membrane and dispersed nuclear chromatin. The characteristic finding was a perichromatin halo delineated by a rim of dense cytoplasm, beyond which a second zone of uniform clearing was observed, similar to changes noted in vulvar intraepithelial neoplasia. There was an absence, however, of either nuclear atypia in the lower cell layers or apoptosis and mitotic arrest observed in vulvar intraepithelial neoplasia and podophyllin therapy. Two (50%) of four lesions tested positive for human papillomavirus nuclei acids, one by polymerase chain reaction and the other by DNA-DNA in situ hybridization. No common human papillomavirus types were found. Pseudobowenoid change might represents a distinct type of vulvar condyloma, perhaps related to an unusual human papillomavirus. PMID- 8729977 TI - 17 beta-hydroxysteroid dehydrogenase in common epithelial ovarian tumors. AB - Type 1 17 beta-hydroxysteroid dehydrogenase (17 beta-HSD) is an estrogen metabolizing enzyme that catalyzes the conversion of estrone to the more biologically potent estradiol. We examined the immunolocalization of 17 beta-HSD in five specimens of normal human ovary and in 51 specimens of common epithelial tumors of the ovary to study the expression of 17 beta-HSD in these ovarian neoplasms. 17 beta-HSD immunoreactivity was detected in granulosa cells of dominant preantral follicles but not in the surface epithelium of the normal cycling human ovary. In 12 patients, the enzyme was not expressed in tumor cells of benign cystadenoma. 17 beta-HSD was immunolocalized in the cytoplasm of four specimens (50%) of eight of tumors of low malignant potential and in 20 specimens (67%) of 30 of invasive carcinoma. 17 beta-HSD immunoreactivity was also observed in nonluteinized stromal cells of mucinous tumors (two adenomas of six, three tumors of five with low malignant potential, and two of 10 invasive carcinomas) but not in other histologic types. Type 1 17 beta-HSD expression, therefore, correlated with malignant transformation of the surface epithelium of the human ovary, supporting a possible role in its progression by increasing in situ estradiol concentration. PMID- 8729978 TI - p53 expression in placentas with hydropic change and hydatidiform moles. AB - Hydatidiform moles result from abnormal fertilization and have been divided into partial and complete forms based on morphologic, cytogenetic, and clinical features. Little is known about their pathogenesis or malignant transformation. We applied an immunohistochemical marker for the p53 tumor suppressor gene product to placentas with hydropic change and hydatidiform moles to determine whether abnormal p53 gene product accumulation occurs in molar gestations. Ploidy of these placentas was determined by flow cytometry and fluorescence in situ hybridization. The mean percentages of p53-positive cells was determined by counting 200 cytotrophoblastic and proliferating trophoblastic cells. The staining intensity was graded on a scale of 1+ (faint) to 3+ (strong). The mean percentage of p53-positive cells for the placentas were as follows: 8.9% +/- 10.5 for hydropic change; 28.0% +/- 13.2 for partial mole; and 41.0% +/- 19.6 for complete mole. There was a significant difference in p53 expression between hydropic change and partial mole (P = 0.05) and hydropic change and complete mole (P = 0.0008). Although there was a difference between partial mole and complete mole, this did not reach statistical significance (P = 0.15). Hydatidiform moles exhibited 2+ to 3+ staining intensity, whereas hydropic placentas exhibited weaker intensity (1-2+). The finding of p53 gene product overaccumulation in partial and complete moles suggests that p53 gene mutations or alternatively, post-transcriptional changes in the p53 gene product occur resulting in inactivation and stabilization of the protein. This may play a role in uncontrolled trophoblastic proliferation and neoplastic transformation. PMID- 8729979 TI - Small cell carcinoma of the uterine cervix with syndrome of inappropriate antidiuretic hormone secretion. AB - In a 59-year-old woman suffering from the syndrome of inappropriate antidiuretic hormone secretion, a small cell carcinoma of the uterine cervix was detected. The tumor was immunoreactive for antidiuretic hormone as well as for neuron specific enolase, chromogranin A, and Leu-7, but not vimentin. Electron microscopic examination of the tumor revealed neurosecretory granules. To our knowledge, this is only the second report of the syndrome of inappropriate antidiuretic hormone secretion with small cell carcinoma of the uterine cervix and the first one confirmed immunohistopathologically. PMID- 8729980 TI - bcl-2 expression and the development of endometrial carcinoma. AB - The bcl-2 gene product inhibits programmed cell death (apoptosis). The expression of this protein has been examined in normal endometrium and found to be cycle dependent with consistent expression in the proliferative phase. In the current report, bcl-2 gene expression was examined in 88 endometrial biopsies that showed 99 histologic patterns ranging from proliferative endometrium to carcinoma. Whereas expression was always detected in 17 samples of proliferative endometrium, eight cases of simple hyperplasia, and five cases of complex hyperplasia, expression was detected in only five (42%) of 12 atypical hyperplasias and in only 27 (47%) of 57 carcinomas. In atypical hyperplasia and carcinoma showing expression, both staining intensity and distribution were less than that seen in their benign counterparts. Three of five biopsies showing both benign endometrium and carcinoma showed positivity in benign but not in malignant glands. In endometrial carcinoma, bcl-2 expression did not correlate with grade, stage, or survival. In addition, in cases of carcinoma, a negative correlation between bcl-2 and p53 expression, which has been reported in other tumors, was not observed in this study. The invariable expression of bcl-2 in proliferative endometrium suggests that this protein may be important for cell survival. Its role in preventing cell death, however, appears to be frequently bypassed in atypical hyperplasia and endometrial carcinoma possibly by other factors impeding programmed cell death. Thus, if an apoptotic pathway usually blocked by bcl-2 expression would not necessarily lead to cell death. Defects in an apoptotic pathway such as the transforming growth factor-beta pathway, which can be blocked by bcl-2, may possibly account for the phenomenon observed in this study. PMID- 8729981 TI - Protocol for immunocytochemical detection of SIL in cervical smears using MIB-1 antibody to Ki-67 [corrected]. AB - Ki-67 nuclear antigen is expressed in upper epithelial levels of intraepithelial neoplasia of the cervix and vulva, variably in condyloma, and in basal and parabasal cells of normal squamous mucosa in histologic preparations. The application of antibodies to Ki-67 as a marker of squamous intraepithelial lesions in cervical smears was explored using either air-dried, acetone-fixed cervical smears obtained from 106 consenting patients or a single slide from archival two-slide cases of squamous intraepithelial lesions MIB-1 monoclonal antibody to Ki-67 was tested using two immunocytochemical techniques. In one set of smears, avidin-biotin peroxidase was used for detection and diaminobenzidine with H2O2 as the chromogen. Some specimens were incubated with 0.3% H2O2 and phosphate buffered saline for blockade of endogenous peroxidase. Alternatively, other air-dried smears were stained using alkaline phosphatase antialkaline phosphatase for detection and new fuchsin as the chromogen. Nuclear staining in squamous intraepithelial lesions was identified in air-dried smears using all of the above methods. Slides stained with avidin-biotin peroxidase and blocked with 0.3% H2O2 and phosphate buffered saline showed less background staining from neutrophils and erythrocytes compared with those without blocking. Slides stained using alkaline phosphatase antialkaline phosphatase showed excessive cytoplasmic staining of endocervical cells, making intraepithelial difficult. No nuclear staining of squamous intraepithelial lesions was observed in destained archival smears. Air-dried smears blocked with 0.3% H2O2 and phosphate buffered saline, incubated with MIB-1, and stained using avidin-biotin peroxidase gave the best results for identification of Ki-67 expression in squamous intraepithelial lesions. PMID- 8729982 TI - Adenoid cystic ovarian carcinoma compared with other adenoid cystic carcinomas of the female genital tract. AB - Adenoid cystic carcinoma (ACC) of the ovary is an extremely rare neoplasm referred to infrequently in the literature. A new case of a pure, primary, ovarian ACC is reported. The patient was a 45-year-old white woman who had a stage IA, well-circumscribed, unilateral, ovarian mass. Histologically, the tumor was composed of small, uniform, and basaloid cells exhibiting predominantly a tubular pattern, as well as demonstrating cribriform and focal solid areas. There was no necrosis and minimal nuclear pleomorphism; the mitotic rate was fewer than three cells per 10 high-power fields. Immunohistochemical examination showed that the tumor cells were strongly positive for cytokeratins CAM 5.2 and AE1/3 and carcinoembryonic antigen. The cells at the periphery of the epithelial aggregates in the cribriform and tubular areas were positive for muscle-specific actin and, occasionally, for S100 protein. Electron microscopic examination revealed that these cells had convoluted nuclei and filamentous structures associated with dense bodies. These findings support a myoepithelial component. A characteristic feature was the production of periodic acid-Schiff stain-positive basement membrane material, which appeared as reduplicated basal lamina by electron microscopic examination. The patient is currently alive, without evidence of recurrence or persistent tumor 7 years after surgical excision. The tumor demonstrated immunohistochemical and ultrastructural findings similar to those of ACCs of salivary gland origin and to other ACCs of the female genital tract. The histogenesis of most ovarian ACCs is through metaplasia of the celomic epithelium, but pure cases might also be of teratomatous origin. PMID- 8729983 TI - Cyclin D1 gene expression in human cervical neoplasia. AB - Amplification of chromosome 11q13 leads to overexpression of a G1 cyclin gene, cyclin D1 (PRAD-1, CCND-1), in many non-cervical human carcinomas. Homology between cyclin D1 and human papillomavirus oncoprotein E7 binding sites for the retinoblastoma tumor-suppressor protein suggests that human papillomavirus oncoproteins cyclin D1, and other cell cycle regulatory proteins may act through a common mechanism in the pathogenesis of human cervical squamous cell carcinoma. We have examined 48 cases of cervical neoplasia by RNA-mRNA in situ hybridization for human papillomavirus mRNA and cyclin D1 mRNA and by immunohistochemistry for cyclin D1 protein expression. Hybridization demonstrated human papillomavirus RNA in all 48 cases with types 6, 16, or 18 in 2, 26, and 20 cases, respectively. Immunohistochemical detection using anti-cyclin D1 rabbit polyclonal antibody 19 demonstrated appropriate cyclin D1 expression at constitutive low levels in normal squamous epithelium and low-grade intraepithelial lesions. Immunohistochemical staining failed to demonstrate significant protein expression in any of the high-grade or invasive lesions. In contrast to the immunohistochemical results, in situ hybridization demonstrated cyclin D1 mRNA overexpression in three of five cases of low-grade squamous intraepithelial lesion, one of eight cases of high-grade squamous intraepithelial lesion, 14 of 18 cases of invasive squamous cell carcinoma, two of five cases of adenocarcinoma in situ, one of seven cases of invasive adenocarcinoma, and two of five cases of small cell undifferentiated carcinoma. Transcriptional activation of cyclin D1 can occur in vivo in human papillomavirus-associated invasive cervical carcinoma, but it does not seem to result in a steady state, increased level of cyclin D1 protein expression. These data indicate a limited role for cyclin D1 protein in the pathogenesis of human papillomavirus-associated invasive cervical squamous carcinoma. They support a model in which human papillomavirus proteins can circumvent cellular requirements for cyclin D1 in human cervical neoplasia. PMID- 8729984 TI - Cytokeratins 7 and 20 and carcinoembryonic antigen in ovarian and colonic carcinoma. AB - The histologic distinction between adenocarcinoma primary to the ovary and metastatic to the ovary can be difficult. In an effort to facilitate this distinction, we have evaluated the use of immunohistochemical techniques with antibodies to cytokeratins 7 and 20, along with antibodies to carcinoembryonic antigen. We studied 24 primary ovarian adenocarcinomas, 11 colonic adenocarcinomas metastatic to the ovary, and 10 primary adenocarcinomas of the colon. Four ovarian adenocarcinomas metastatic to the colon were also studied. All but one of the primary and metastatic colonic carcinomas were negative for cytokeratin 7, whereas all the primary and metastatic ovarian carcinomas were positive for cytokeratin 7. The tumors metastatic to the ovary were all positive for cytokeratin 20 and carcinoembryonic antigen. Among the primary ovarian carcinomas, none of six serous tumors, three of seven endometrioid tumors, and three of 11 mucinous tumors were positive for cytokeratin 20. Ten of the primary ovarian tumors were negative for carcinoembryonic antigen using both monoclonal and polyclonal antibodies. One of the ovarian tumors was negative for carcinoembryonic antigen with the monoclonal antibody but positive using the polyclonal antibody. Cytokeratin 7 is the most helpful marker in the distinction between primary ovarian carcinoma and colonic carcinoma metastatic to the ovary. PMID- 8729985 TI - bcl-2 is down-regulated in atypical endometrial hyperplasia and adenocarcinoma. AB - The bcl-2 protein, which protects cells from apoptosis, is normally expressed in a number of adult tissues. Dysregulated bcl-2 expression, secondary to (14;18) chromosomal translocation, seems to promote the development of follicular lymphomas, and recent findings of bcl-2 protein in several solid tumors suggest that it might contribute to the genesis of many other neoplasms. bcl-2 is also highly expressed in normal proliferative endometrium and markedly down-regulated in secretory endometrium, which suggests that its expression is estrogen regulated. Because the development of most endometrial carcinomas is associated with hyperestrogenic states, we began the investigation of the role of bcl-2 in endometrial carcinogenesis by immunohistochemically quantifying its expression in proliferative, hyperplastic, atypically hyperplastic, and carcinomatous endometrium. The results of this study show that bcl-2 is relatively highly expressed in proliferative (n = 11) and hyperplastic (n = 18) endometrium, with respective mean staining scores of 3.59 and 3.47 (scale, 0-4), but is significantly (P < 0.001) down-regulated in atypical hyperplasia (n = 11; score, 0.82), and adenocarcinoma (n = 34; score, 0.86). bcl-2 expression did not correlate with stage, grade, estrogen-receptor, or progesterone-receptor expression. Polymerase chain reaction analyses of DNA isolated from several endometrial carcinomas were negative for (14;18) translocation involving the bcl 2 gene. Thus, bcl-2 apparently plays no role in the progression of atypical hyperplasia to carcinoma or in the development of high-grade or advanced-stage endometrial carcinoma. These results, however, do not rule out the involvement of bcl-2 in very early, preatypical hyperplasia phases of endometrial carcinogenesis. Finally, the marked difference in bcl-2 expression in hyperplastic and atypically hyperplastic glands might prove to be diagnostically useful in the often difficult distinction of these entities. PMID- 8729986 TI - Expression of bcl-2 by breast cancer: a possible diagnostic application. AB - Expression of bcl-2 is most commonly associated with the t(14;18) translocation present in most folicular lymphomas (1). More recently, bcl-2 oncoprotein has been identified in normal tissues and in nonhematologic malignancies. In this study, we investigate the use of bcl-2 as a marker to distinguish metastatic breast carcinoma from primary lung and gastric cancers, and we evaluate the role of bcl-2 as an independent prognostic factor in breast carcinoma and its relationship to other breast cancer markers. bcl-2 immunostains were done on 371 adenocarcinomas of the breast, lung, and stomach. Additionally, 231 samples of metastases from patients with breast or gastric cancer were evaluated for bcl-2 expression. All breast cancer tissue samples had immunohistochemical data on expression of estrogen and progesterone receptors, p53, neu/cerb2, and MIB-1. A large proportion (79.3%) of invasive breast carcinomas expressed bcl-2, whereas only 5.6% and 8.3% of pulmonary and gastric carcinomas did. Moreover, staining was moderate to intense in 70.2% of the breast cancers, compared with only one specimen of lung carcinoma (1.9%) and gastric carcinoma (0.9%) that showed moderate staining. There was agreement of bcl-2 expression between primary and metastatic sites in all specimens except one. Expression of bcl-2 in breast adenocarcinomas was significantly associated with hormone receptor positivity and low histologic grade. Nonetheless, 20.6% of bcl-2-positive specimens were estrogen receptor negative and 24.2% of bcl-2-positive specimens were progesterone receptor negative. Neither the presence nor the absence of bcl-2 expression significantly predicted disease-free survival or overall survival in patients with breast cancer. We conclude that adenocarcinomas with intense bcl-2 staining are more likely to be of breast than of pulmonary or gastric origin. We recommend the addition of bcl-2 to a panel of antibodies (estrogen receptor, GCDFP-15, and S100) that might contribute to the identification of a larger proportion of metastatic breast carcinomas, because almost one-half of the estrogen-receptor negative cancers were bcl-2 positive. PMID- 8729987 TI - Surfactant proteins and thyroid transcription factor-1 in pulmonary and breast carcinomas. AB - Antibodies to the pulmonary epithelial cell-specific proteins surfactant proteins A and B (SP-A and SP-B) and to thyroid transcription factor-1 (TTF-1), a homeodomain nuclear transcription protein, were used as immunohistochemical markers to asses their ability to distinguish primary pulmonary non-small cell carcinomas (n = 57) from carcinomas of the breast (n = 51). SP-A, SP-B, and TTF-1 were detected in 49%, 53%, and 63% of non-small cell carcinomas, respectively. These three antibodies stained pulmonary adenocarcinomas in 54%, 63% and 76% of specimens, respectively. Squamous cell carcinomas rarely stained using these markers. Antibodies to SP-B and TTF-1 never stained any of the 51 breast carcinomas, whereas four of these tumors stained for SP-A. To better define the potential diagnostic value of these antibodies, 13 breast carcinomas metastatic to the lung were studied. Of the three antibodies tested, only TTF-1 seemed useful, because none of the 13 metastatic tumors showed immunoreactivity to this antibody, whereas six specimens (46%) showed reactivity for both SP-A and SP-B. To emphasize further the potential usefulness of antibodies to TTF-1, sections of adenocarcinomas of the colon (n = 18) and prostate (n = 9), renal cell carcinomas (n = 8), and epithelioid mesotheliomas (n = 4) were evaluated; none was positive. Only one of 66 gastric and one of eight endometrial adenocarcinomas showed focal positivity. These results demonstrate the usefulness of immunodetection of a pulmonary cell selective transcription protein (TTF-1) in the diagnosis of pulmonary adenocarcinoma, readily distinguishing breast carcinomas from primary pulmonary adenocarcinomas. In contrast, staining for SP-A and SP-B is of limited value, because there is an unacceptably high rate of cross-reactivity between breast carcinomas metastatic to the lung and primary pulmonary carcinomas. The latter finding illustrates and supports the fact that tumor marker phenotypes might differ in primary and secondary tissue sites. PMID- 8729988 TI - Recommendations for reporting resected prostatic carcinomas. Association of Directors of Anatomic and Surgical Pathology. PMID- 8729989 TI - Correspondence re: Mills SE, Kempson RL, Fechner RE, et al.: Guardians of the wax ... and the patient. Mod Pathol 8:699, 1995. PMID- 8729990 TI - Correspondence re: Fleischhacker M, Lee S, Spira S, Takeuchi S, Koeffler HP: DNA aneuploidy in morphologically normal colons from patients with colon cancer. Mod Pathol 8:360, 1995. PMID- 8729991 TI - Correspondence re: Thomas PA, Cangiarella J Raab SS, Waisman J: Fine needle aspiration biopsy of proliferative breast disease. Mod Pathol 8:130, 1995. PMID- 8729992 TI - Correspondence re: Lavoie M, Morency RM: Low-grade papillary adenomatous tumors of the temporal bone: report of two cases and review of the literature. Mod Pathol 8:603, 1995. PMID- 8729993 TI - The changing role of mechanical ventilation in COPD. PMID- 8729994 TI - Lymphangioleiomyomatosis and tuberous sclerosis: where is the border? PMID- 8729995 TI - Extrathoracic angiomyolipomas in lymphangioleiomyomatosis. AB - Pulmonary lymphangioleiomyomatosis (LAM) is a rare disorder that affects women and can lead to serious respiratory impairment. Since Bourneville's tuberous sclerosis (TS) was first reported, the striking similarities between the two entities have led many to believe that LAM is a forme fruste of TS. This is suggested by reports that angiomyolipomas, rare tumours in themselves, are reported in 40-80% of TS patients and occur in 15-30% of LAM patients. A retrospective chart review was conducted of 14 patients that presented to our institution with a diagnosis of LAM. We sought to document the clinical manifestations, particularly the incidence and location of extrathoracic tumours, in order to further support the hypothesis that LAM is a forme fruste of TS. Twelve patients had premenopausal onset of symptoms and two postmenopausal. The diagnosis was confirmed histologically (n = 12) and/or by computed tomography (CT) scan of the thorax (n = 12). Imaging investigations revealed extrathoracic tumours in 12 of 14 patients (86%). Eight of the 14 patients (57%) had renal tumours consistent with angiomyolipomas (bilateral in five patients). Only one patient had renal symptoms (flank pain and haematuria). All had normal serum creatinine, one had a reduced creatinine clearance. Extrathoracic nonrenal tumours were discovered in the pancreas, adrenals and uterus, findings previously unreported in LAM. In summary, the incidence of extrathoracic tumours in lymphangioleiomyomatosis patients is much higher than previously reported in the literature. This increased association supports the theory that lymphangioleiomyomatosis and tuberous sclerosis represent part of a spectrum of a similar disease process. PMID- 8729996 TI - Expiratory chest radiographs do not improve visibility of small apical pneumothoraces by enhanced contrast. AB - Demonstration of small apical pneumothoraces is supposed to be facilitated by expiratory chest radiographs. This study aimed to analyse the assumed enhancement of visual contrast on expiratory chest radiographs in patients with small apical pneumothoraces. Optical densities (OD) were obtained with a densitometer (X-rite 3001) on 54 paired inspiratory and expiratory chest radiographs of 22 consecutive patients with small apical pneumothoraces. The ODs were measured: at the intervertebral space between the first and second thoracic vertebrae (Area 1); at the peripheral part of the affected lung parenchyma (Area 2); and at the adjacent intrapleural space (Area 3). Excellent correlations of OD of each area were obtained between paired inspiratory and expiratory chest radiographs. The ODs of all areas on expiratory chest radiographs were significantly higher than on inspiratory chest radiographs. Contrast between pulmonary parenchyma and intrapleural air in inspiratory and expiratory films did not differ significantly. Expiratory chest radiographs do not improve visibility of small apical pneumothoraces by enhanced contrast between pulmonary parenchyma and intrapleural air. Expiratory chest radiographs show equally increased OD in the area of lung tissue and intrapleural air, caused by increased extrapulmonary tissue density during expiration, resulting in increased radiation exposure monitored by the ionization chambers of standard radiological equipment. If expiratory chest radiographs are really improving the visibility of apical pneumothoraces, there must be other reasons than contrast enhancement to explain this. PMID- 8729997 TI - Evaluation of the solitary pulmonary nodule by positron emission tomography imaging. AB - Current noninvasive imaging methods are not sufficiently reliable for accurate detection of malignancy in most solitary pulmonary nodules (SPNs). Positron emission tomography (PET) using 18-fluorodeoxyglucose (FDG), showing increased FDG uptake and retention in malignant cells, has proved useful to differentiate malignant from benign tissue and could, therefore, contribute to the evaluation of the SPN. We performed a prospective study of 50 patients referred to the Pneumology Department with unclear diagnoses of SPN after conventional radiological screening. PET study was performed on each subject before an invasive procedure was proposed. Thirty three patients had a malignant nodule and 17 had a benign nodule. The mean size of malignant nodule was 3 cm (range 1.5-4.5 cm). All showed a marked increase in 18-FDG uptake. The mean size of benign nodule was 1.8 cm (range 0.5-3.5 cm). PET imaging showed the absence of 18-FDG uptake and correctly identified 15 of 17 benign nodules. There was two false positive cases with a moderate increase in 18-FDG uptake (1 postprimary tuberculosis; and 1 anthracosilicotic nodule with nonspecific inflammation). At present, the sensitivity and specificity of the method are 100 and 88%, respectively. The positive and negative predictive values of PET imaging for SPNs are 94 and 100%, respectively. Our preliminary results demonstrate that PET-FDG imaging is a noninvasive technique, which appears highly accurate in differentiating malignant SPN from benign SPN. PMID- 8729998 TI - Pulmonary function and exercise capacity after lung resection. AB - The influence of pulmonary resection on functional capacity can be assessed in different ways. The aim of this study was to compare the effect of lobectomy and pneumonectomy on pulmonary function tests (PFT), exercise capacity and perception of symptoms. Sixty eight patients underwent functional assessment with PFT and exercise testing before (Preop), and 3 and 6 months after lung resection. In 50 (36 males and 14 females; mean age 61 yrs) a lobectomy was performed and in 18 (13 males and 5 females; mean age 59 yrs) a pneumonectomy was performed. Three months after lobectomy, forced vital capacity (FVC), forced expiratory volume in one second (FEV1), total lung capacity (TLC), transfer factor of the lungs for carbon monoxide (TL,CO) and maximal oxygen uptake (V'O2,max) were significantly lower than Preop values, increasing significantly from 3 to 6 months after resection. Three months after pneumonectomy, all parameters were significantly lower than Preop values and significantly lower than postlobectomy values and did not recover from 3 to 6 months after resection. At 6 months after resection significant deficits persisted in comparison with Preop: for FVC 7% and 36%, FEV1 9% and 34%, TLC 10% and 33% for lobectomy and pneumonectomy, respectively; and V'O2,max 20% after pneumonectomy only. Exercise was limited by leg muscle fatigue in 53% of all patients at Preop. This was not altered by lobectomy, but there was a switch to dyspnoea as the limiting factor after pneumonectomy (61% of patients at 3 months and 50% at 6 months after resection). Furthermore, pneumonectomy compared to lobectomy led to a significantly smaller breathing reserve (mean +/- SD) (28 +/- 13 vs 37 +/- 16% at 3 months; and 24 +/- 11% vs 33 +/- 12% at 6 months post resection) and lower arterial oxygen tension at peak exercise 10.1 +/ 1.5 vs 11.5 +/- 1.6 kPa (76 +/- 11 vs 86 +/- 12 mmHg) at 3 months; 10.1 +/- 1.3 vs 11.3 +/- 1.6 kPa (76 +/- 10 vs 85 +/- 12 mmHg) at 6 months postresection. We conclude that measurements of conventional pulmonary function tests alone overestimate the decrease in functional capacity after lung resection. Exercise capacity after lobectomy is unchanged, whereas pneumonectomy leads to a 20% decrease, probably due to the reduced area of gas exchange. PMID- 8729999 TI - Noninvasive mechanical ventilation improves the immediate and long-term outcome of COPD patients with acute respiratory failure. AB - Noninvasive positive pressure ventilation (NPPV) has been proposed in COPD patients with acute on chronic respiratory failure (ACRF) in order to avoid endotracheal intubation and to improve immediate outcome, but long-term outcome of this therapeutic approach is still undefined. We evaluated short- and long term (1 year) outcome of early administration of NPPV in 24 patients with ACRF due to exacerbated COPD (Group A) in comparison with 24 matched historical control patients treated conventionally (Group B). Patients of Group A were initially treated with NPPV via nasal mask in the presence of pH < or = 7.32, and/or Pa,O2 < 7.98 kPa, and/or Pa,CO2 > 7.18 kPa, plus signs of respiratory distress. In-hospital survival rate was not significantly different in Group A vs Group B, but the patients treated with NPPV showed an earlier improvement in blood gases and a better pH and respiratory rate at discharge. Only 2 patients of Group A needed endotracheal intubation as compared with 9 of Group B. Hospital stay was significantly reduced in survivors of Group A vs Group B. Further severe relapses of ACRF in Group A were treated using NPPV. The number and length of further hospitalizations for pulmonary exacerbations were significantly higher in Group B compared with Group A. The survival rate at 12 months was significantly lower in Group B than in Group A (50% vs 71%). In conclusion, NPPV administration in patients with ACRF due to exacerbated COPD improves not only immediate but also long-term outcome. PMID- 8730000 TI - Variables related to increased mortality following out-patient pulmonary rehabilitation. AB - Although patients with advanced pulmonary diseases have significant improvement in exercise ability and functional status following comprehensive out-patient pulmonary rehabilitation (OPR), their long-term prognosis once they have reached this stage of their diseases remains poor. To further evaluate predictors of increased mortality in these patients, we related patient characteristics and short-term outcome obtained during OPR assessment of 158 patients to subsequent survival. The time period from OPR to death or collection of survival data was 40.0 +/- 17.1 months. The following variables were tested individually and in stepwise fashion using a proportional hazards model: 1) age; 2) gender; 3) pulmonary diagnosis; 4) prebronchodilator forced expiratory volume in one second (FEV1); 5) arterial oxygen tension (Pa,O2) and arterial carbon dioxide tension (Pa,CO2); 6) body mass index (BMI); 7) pre- and post-OPR 12 min walking distance (12-MW); 8) pre- and post-OPR quality of life, using the Chronic Respiratory Disease Questionnaire (CRDQ); 9) number and type of nonpulmonary diagnoses; and 10) number of medications. Separate survival analyses were performed for all deaths (the total group), respiratory deaths only (nonrespiratory deaths excluded), and nonrespiratory deaths only (respiratory deaths excluded). Forty three patients (27%) died during the study period; and the 3 year survival was 80%. For all three survival analyses, the post-OPR 12-MW was the most significant variable related to prognosis: patients with low timed walking distance had increased mortality both from respiratory and nonrespiratory causes. Other variables related to increased mortality included: elevated Pa,CO2; low pre-OPR 12-MW; reduced Pa,O2; low FEV1; low BMI, increased number of medications, and increased CRDQ dyspnoea. These results indicate that the timed walking distance following out-patient pulmonary rehabilitation is an important predictor of survival in patients with advanced pulmonary disease. PMID- 8730001 TI - Long-term effects of therapy on respiratory health. AB - Only scant information is available on the long-term consequences to respiratory health of treatment with bronchodilators and oral corticosteroids. In the present study, we aimed to gain more information about these consequences. We examined 712 men working in the Paris area, by means of a subjective assessment of whether their respiratory health worsened or improved from 1960 to 1972, the decline in forced expiratory volume in one second (FEV1) over this time-period, and mortality from 1972 to 1992, in relation to respiratory therapy dispensed during a 2 year period in 1970-1971, as recorded in social security reimbursement records. As expected, subjects with respiratory symptoms or airflow limitation were more likely to have been prescribed respiratory therapy. After accounting for the effect of lung function level and smoking, subjects dispensed inhaled beta-agonists were likely to feel their condition had worsened and had a greater decline in FEV1 from 1960 to 1972. Among subjects with airflow limitation who reported asthma or persistent wheeze, having been dispensed oral corticosteroids on an intermittent basis was associated with improved survival (relative risk (RR) 0.32; 95% confidence interval (95% CI): 0.10-0.91) after adjusting for FEV1 level and smoking category. Our results add to the evidence that regular use of beta-agonist bronchodilators may be associated with adverse effects on respiratory health, whilst intermittent use of corticosteroids may be of long term benefit. PMID- 8730002 TI - Integrated care for asthma: matching care to the patient. AB - The purpose of the present study was to investigate whether criteria associated with assignment of asthma patients between general practice (GP) care alone, integrated care (shared between GP care and hospital clinic) or conventional specialist review could be identified, and whether outcomes for these patients differed over the next 12 months. Seven hundred and sixty four patients with a diagnosis of asthma and previously assigned to either integrated care or clinic care were reviewed after 1 year and reassigned. These patients were then followed for another 12 months and clinical data were collected over this time. After 12 months in clinic care or integrated care, assignment to integrated care was predicted by previous participation in integrated care (OR 2.94), patient preference for integrated care (OR 3.7), no admission (OR 1.56), fewer steroid courses during the previous year (OR 0.88) and nonattendance at review (OR 0.43) in the previous 12 months. Patient discharge to GP care was predicted by higher level of forced expiratory volume in one second (FEV1) (OR 1.49), lower number of GP consultations for troublesome asthma (OR 0.78), and nonattendance for review in the preceding year (OR 2.15). In the following 12 months, the three groups of patients differed significantly in hospital admissions (Discharged = 0.008; Integrated = 0.12; Clinic = 0.31), bronchodilators prescribed (Discharged = 8.5; Integrated = 10.2; Clinic = 13.9), GP consultations (Discharged = 1.3; Integrated = 3.0; Clinic = 4.1) and oral steroid courses (Discharged = 0.62; Integrated = 1.7; Clinic = 2.4). Patients assigned to integrated care, clinic care or discharged to general practice care form three distinct patient populations differing retrospectively and prospectively in morbidity and admission risk. In particular, patients assigned to integrated care fall midway in risk and morbidity between those discharged or those retained in clinic care. These results suggest that integrated care provides general practitioners with a system of management for asthma patients, for whom they do not wish frequent specialist review but who they do not believe can safely be discharged to general practice care only. PMID- 8730003 TI - Effect of salmeterol compared with beclomethasone on allergen-induced asthmatic and inflammatory responses. AB - Salmeterol is a selective long-acting beta 2-agonist bronchodilator considered to have added anti-inflammatory effects, but this is controversial. We investigated the effects of a single dose of salmeterol, 100 micrograms, on the physiological and inflammatory responses to inhaled allergen and compared these with the effects of a single dose of beclomethasone, 500 micrograms, and of placebo. Eight atopic adults with mild stable asthma, treated only with inhaled short-acting beta 2-agonist when needed, attended the laboratory sequentially for screening tests, two single-blind control inhalation tests preceded 30 min by placebo or salmeterol and three allergen inhalation tests preceded by placebo, salmeterol or beclomethasone double-blind in random order. Airway responsiveness to methacholine (assessed as the provocative concentration of methacholine producing 20% fall in forced expiratory volume in one second (PC20)), induced sputum eosinophils, blood eosinophils and serum eosinophil cationic protein (ECP) were examined before and 7-48 h after treatment. The statistical power to detect twofold changes in blood and sputum parameters was > or = 90%. Salmeterol inhaled before allergen challenge completely prevented the early asthmatic response, late asthmatic response and fall in methacholine PC20 at 24 h, and produced additional bronchodilatation. These effects were similar to those obtained by the inhalation of a single dose of salmeterol before the control inhalation test, and significantly better than those observed after a single dose of beclomethasone inhaled before the allergen test. Beclomethasone had no effect on the early asthmatic response or on the fall in methacholine PC20 at 24 h but partially inhibited the late asthmatic response. Neither salmeterol nor beclomethasone had any significant effect on sputum or blood inflammatory changes 7-48 h after allergen inhalation. In conclusion, whilst salmeterol had no demonstrable anti inflammatory action in sputum after allergen challenge in asthma, neither did a single dose of the positive anti-inflammatory control, beclomethasone. The latter result excludes a more positive judgement on the possible anti-inflammatory action of salmeterol. However, the results do indicate that potent functional effects of a single dose of salmeterol can mask the airway inflammatory cell influx caused by inhaled allergen. PMID- 8730004 TI - Low-dose theophylline modulates T-lymphocyte activation in allergen-challenged asthmatics. AB - Theophylline has been shown by several investigators to attenuate the late asthmatic response (LAR) to inhaled allergen, suggesting that it has anti inflammatory or immunomodulatory properties. We have, therefore, undertaken a double-blind, placebo-controlled study to examine the effects of low-dose theophylline on bronchoalveolar lavage (BAL) and blood T-lymphocyte profile and activation in asthmatics following antigen challenge and the development of a LAR. Peripheral blood and BAL samples were obtained from 17 subjects with mild atopic asthma before and after 6 weeks of treatment with either oral theophylline or placebo. The mean serum theophylline concentration achieved was 6.6 micrograms.mL-1, which is below the currently accepted therapeutic range. Following theophylline therapy, there was a significant decrease in the number of BAL lymphocytes compared to placebo. On flow cytometric analysis of BAL cells, a significant loss of CD3+ T-lymphocytes, comprising both CD4+ and CD8+ subsets, was demonstrated. Moreover, there was a decrease in the number of BAL CD4+ T cells expressing the activation marker very late activation antigen-1 (VLA-1), and an apparent reduction in human leucocyte antigen-DR (HLA-DR). Correspondingly, this was accompanied in the blood by an elevation in the proportion of activated CD4+ T-lymphocytes, in particular those expressing HLA DR. These findings provide further evidence that theophylline has an anti inflammatory action in asthma. PMID- 8730005 TI - Modulation of ICAM-1 expression in human alveolar macrophages in vitro. AB - Modulation of intercellular adhesion molecule-1 (ICAM-1) expression may be a basic mechanism by which alveolar macrophages (AMs) regulate the inflammatory process in the lung in response to local stimuli. As a model for studying the anti-inflammatory activity of drugs on human AMs, we investigated the effects of fusafungine, an antibiotic for local use by aerosol with anti-inflammatory properties, and that of the glucocorticoid dexamethasone, on ICAM-1 expression induced in vitro by recombinant interferon-gamma (rIFN-gamma). ICAM-1 protein expression was studied on AMs by means of flow cytometry with an anti-CD54 monoclonal antibody; messenger ribonucleic acid (mRNA) levels were determined by reverse transcriptase-polymerase chain reaction (RT-PCR). ICAM-1 was expressed before culture on 21% of bronchoalveolar lavage (BAL) cells, with low intensity. Culture for 24 h with rIFN-gamma resulted in a significant increase in ICAM-1 protein expression (82% of cells were strongly positive). Fusafungine significantly inhibited rIFN-gamma-induced ICAM-1-protein expression on AMs in a concentration-dependent fashion. The mechanism of ICAM-1 downregulation was mainly post-transcriptional, but also partly transcriptional. By contrast, dexamethasone did not influence rIFN-gamma-induced ICAM-1 expression. This in vitro model using human AMs should prove useful for investigating the cellular and molecular targets of anti-inflammatory drugs. PMID- 8730006 TI - Hyperbaric oxygenation upregulates rat lung Na,K-ATPase. AB - Exposure of rats to hyperoxia is associated with increased active Na+ transport in rat lungs and increased Na,K-adenosine triphosphate (ATPase) expression in alveolar epithelial cells. Hyperbaric oxygenation (HBO) has been reported to act as an accelerated model of hyperoxic cell damage. Sublethal and intermittent exposure to HBO, however, has been suggested to upregulate endogenous protective mechanisms. In the present study, we tested whether short-term HBO, prior to inducing lung injury, would upregulate lung Na,K-ATPase. The results show that HBO, either intermittent or single 2.5 h exposure, increased lung Na,K-ATPase alpha-1 and beta-1 messenger ribonucleic acid (mRNA) transcript levels up to fourfold. Na,K-ATPase activity in lungs of rats exposed to HBO increased twofold during the first 2 h following removal from the hyperbaric chamber, and remained elevated for up to 6 h following HBO. Conceivably, the increase in Na,K-ATPase activity following HBO is due to an increase in activity from a basal to a higher rate, or possibly due to recruitment/translocation of Na,K-ATPases from inner membranes to the plasma membrane. PMID- 8730007 TI - Effects of moguisteine, a peripheral nonnarcotic antitussive agent, on airway inflammation in guinea-pigs in vivo. AB - Cough is a common symptom of respiratory diseases associated with irritation or inflammation of the airways, and symptomatic antitussive drugs are frequently prescribed to control an abnormal cough reflex. Our aim was to evaluate the effects of moguisteine, a novel, peripheral, nonnarcotic antitussive agent, on airway inflammation induced in guinea-pigs with a variety of stimuli. These stimuli included exposure to tobacco smoke for 10 min, to elicit airway hyperreactivity, eosinophil recruitment in bronchoalveolar lavage (BAL), airway epithelial damage and plasma exudation; graded platelet-activating factor (PAF) infusion (600 ng.kg-1 over one h), to induce airway hyperreactivity; 2% ovalbumin (OA) aerosol challenge in 1% OA-sensitized animals, to induce late-phase (17 and 72 h) airway leucocyte accumulation. We also assessed the activity of moguisteine on plasma leakage induced by capsaicin, on bronchoconstriction induced by acetylcholine (ACh), histamine (H) and PAF, and on leukotriene mediated allergic bronchospasm in OA-sensitized guinea-pig. Moguisteine (p.o. and i.m.) and dexamethasone (p.o. and i.m.) dose-dependently reduced tobacco smoke-induced bronchial hyperreactivity. Moguisteine and dexamethasone abolished eosinophil recruitment in BAL, prevented the sloughing of the epithelium and significantly reduced airway microvascular leakage. Both agents were also highly effective in reducing bronchial hyperreactivity elicited by PAF infusion. In addition, moguisteine was active in inhibiting airway neutrophil and eosinophil accumulation in BAL observed 17 and 72 h after OA challenge in sensitized guinea pigs. In contrast to dexamethasone, moguisteine did not prevent capsaicin-induced plasma leakage. It was also ineffective against bronchoconstriction as induced by ACh, H, and PAF and failed to inhibit leukotriene-dependent bronchospasm. Our data suggest that moguisteine represents an antitussive compound endowed with interesting airway anti-inflammatory properties in guinea-pigs in vivo. Its mechanism of action remains to be elucidated. PMID- 8730008 TI - Involvement of endogenous tachykinins in LTD4-induced airway responses. AB - Leukotriene D4-(LTD4) has been reported to cause tachykinin release from airway sensory nerves. However, the functional significance of endogenously released tachykinins in LTD4-mediated airway responses has not been fully clarified. The aim of this study was to investigate whether LTD4-induced airway responses are due, in part, to tachykinin release in guinea-pigs. Airway plasma exudation and bronchoconstriction were assessed by measuring extravasation of Evans blue dye and by mean pulmonary resistance (RL) in the presence of atropine (1 mg.kg-1 i.v.) and propranolol (1 mg.kg-1 i.v.), respectively. LTD4 (5 micrograms.mL-1 for 1 min) inhalation caused increase in plasma exudation and RL. Capsaicin pretreatment of animals to deplete sensory neuropeptides significantly inhibited LTD4-induced plasma exudation in the main bronchi, but not in the central (cIPA) and peripheral intrapulmonary airways (pIPA). Pretreatment with specific tachykinin neurokinin-1 (NK1)-receptor antagonists, FK 888 (10 mg.kg-1 i.v.) and CP 96345 (4 mg.kg-1 i.v.), also significantly reduced LTD4-induced plasma exudation in the main bronchi, and in the main bronchi and cIPA, respectively. However, these antagonists did not significantly affect the LTD4-induced increase in RL. In contrast, neurokinin-2 (NK2)-receptor antagonist, SR 48968 (0.3 mg.kg-1 i.v.), significantly inhibited the bronchoconstriction after LTD4-inhalation. These results suggest that leukotriene D4-induced bronchoconstriction and plasma exudation in guinea-pigs are, in part, due to tachykinin release from airway sensory nerves. PMID- 8730009 TI - Role of interleukin-5 and substance P in development of airway hyperreactivity to histamine in guinea-pigs. AB - In this study, we examined the mechanism by which bronchoalveolar lavage (BAL) cells induced hyperreactivity of the trachea in vitro. As both interleukin-5 (IL 5) and substance P (SP) appeared to be involved, the effect of these mediators was examined in vivo. Tracheae were incubated with BAL cells from ovalbumin or saline challenged animals, and from naive animals, in the absence or presence of either IL-5, SP, or both. In addition, the effect of intra-airway application of IL-5, SP, both, or vehicle on tracheal hyperreactivity was examined. Incubation of tracheae with BAL cells from ovalbumin challenged animals induced an increase (30 +/- 10%) in the maximal response to histamine. The hyperreactivity could be completely inhibited by co-incubation with 5-lipoxygenase inhibitor, AA861. The hyperreactivity could be mimicked by incubation of tracheae with BAL cells from naive animals in the presence of IL-5 and SP. After in vivo administration of either IL-5 or SP, maximal responses to histamine were increased and amounted to 105 +/- 35 and 101 +/- 37%, respectively. Administration of IL-5 but not SP induced a significant increase in the number of eosinophils (67 +/- 22%) and eosinophil peroxidase (EPO) activity (94 +/- 33%) in BAL cells. The simultaneous administration of IL-5 and SP did not potentiate the hyperreactivity and eosinophilia observed with IL-5 alone. These data suggest that IL-5 is important in the recruitment of eosinophils, whereas both IL-5 and substance P are involved in the induction of airway hyperreactivity. PMID- 8730010 TI - Effect of diameter on force generation and responsiveness of bronchial segments and rings. AB - In this study, isovolumic bronchial segments and bronchial rings were used to investigate the influence of airway diameter on smooth muscle force generation and acetylcholine responsiveness. Segments with internal diameters ranging from 1.0-6.0 mm were obtained from the mainstem bronchus of eight pigs. Responses to increasing acetylcholine concentrations were quantified in segments by intralumenal pressure (cmH2O), and in rings by tension (g.cm-1). The negative log of the concentration producing half the maximal effect (EC50) (i.e. pD2) to acetylcholine was calculated for each segment and ring. Ring tension was used to calculate a theoretical lumen pressure for each ring, and this, along with the pD2, was compared with values obtained from segments of the same diameter. Intermediate-sized segments produced significantly greater intralumenal pressures than did large or small segments. Small segments were 160 times more sensitive to acetylcholine than large segments. In contrast to the segments, bronchial rings showed no effect of size on acetylcholine sensitivity. Theoretical ring lumen pressures matched those measured for large and intermediate segments, but not for small segments. The different behaviour of bronchial segments and rings obtained from the same sized airway suggests that the three-dimensional architecture of the airway is an important factor in determining behaviour, particularly in small airways. PMID- 8730011 TI - Evaluation of methacholine dose-response curves by linear and exponential mathematical models: goodness-of-fit and validity of extrapolation. AB - Several models have been proposed to analyse dose-response curves recorded in bronchoprovocation challenge tests. The aims of the present work were: 1) to investigate which model (linear vs exponential) and which minimization method (trials and errors vs Levenberg-Marquardt) gives better results in terms of data interpolation (goodness-of-fit); and 2) to verify the validity of extrapolation by comparing forced expiratory volume in one second (FEV1) observed after 4 mg methacholine with values extrapolated after truncation of the curves at 2 mg. For these purposes, methacholine dose-response curves were obtained in 832 subjects from a random population sample, as part of the European Community Respiratory Health Survey (ECRHS) in Italy. Methacholine was inhaled up to a maximum dose of 6 mg by dosimeter technique. The coefficient of determination (r2) was significantly higher with the exponential model (0.81 +/- 0.22; mean +/- SD) than with the linear model (0.69 +/- 0.27). With both models, extrapolated values were usually lower than observed values. As a consequence, a 20% fall in FEV1 with respect to postsaline FEV1 was observed in only 24% and 21% of the tests, where a 20% fall had been predicted, respectively, according to the linear and exponential model. In conclusion, exponential models are better than linear models with respect to data interpolation of methacholine dose-response curves. However, they are worse with respect to extrapolation to higher doses. With any model, extrapolation of dose-response curves by one doubling-dose should be avoided. PMID- 8730012 TI - Relationship between supramaximal flows and flow-limiting mechanisms. AB - The aim of this study was to evaluate the relationship between supramaximal flows (SF) and indices of airway mechanics. We studied 26 asymptomatic young subjects (13 smokers, mean +/- SD 15.9 +/- 6.6 pack-years. Subjects performed maximal expiratory flow-volume (MEFV) curves, according to the American Thoracic Society (ATS) criteria, on a rolling-seal spirometer and then repeated them through a specially devised valve, which occluded the mouthpiece either 3 or 6 times.s-1 with a ratio of open:closed time of 3:1. Envelope MEFV curves for 3 and 6 Hz occlusions were constructed passing through the SF obtained after each occlusion, and the increment in flow at 50% of vital capacity was measured with respect to the basal curve (delta V' max50). We found that the delta V' max50 at 3 and 6 Hz correlated to the baseline forced mid-expiratory flow (FEF25-75) in % of predicted value (r = -0.73 and r = -0.55, respectively). our results suggest that inhomogeneities within the lung are an important mechanism in the occurrence of supramaximal flow in normal subjects. PMID- 8730013 TI - Fever and leucocytosis accompanying asthmatic reactions due to occupational agents: frequency and associated factors. AB - Fever is sometimes associated with asthmatic reactions following specific inhalation challenges with occupational agents. Our aims were to estimate the prevalence of fever in subjects with occupational asthma confirmed by specific inhalation challenge and to examine the characteristics and clinical correlates of subjects who develop fever on specific inhalation challenge. We performed a retrospective analysis of 317 subjects who had positive specific inhalation challenge to occupational agents and a comparison of subjects who developed fever after specific inhalation challenge with a random sample of those who did not. Fifteen subjects (5%) developed fever associated with positive specific inhalation challenge. They were compared with a random sample of 60 subjects who did not develop fever. The fever group: 1) showed a larger increase in absolute number of blood neutrophils, and a larger decrease both of blood lymphocytes and forced vital capacity after specific inhalation challenge; 2) included fewer atopics; 3) had been exposed more frequently to low rather than high molecular compounds; and 4) always experienced a late reaction and had more atypical reactions. We conclude that after positive specific inhalation challenge; fever occurs infrequently, and is associated with an increase in blood neutrophils and a decrease in blood lymphocytes and forced vital capacity. Such reactions seem more likely to occur in nonatopic subjects exposed to low molecular weight agents. PMID- 8730014 TI - Pig farmers have signs of bronchial inflammation and increased numbers of lymphocytes and neutrophils in BAL fluid. AB - The purpose of this study was to investigate whether pig farmers had inflammation of the bronchial mucosa and activation of bronchoalveolar lavage (BAL) cells. Pig farmers are exposed to high dust levels and have a prevalence of work-related respiratory symptoms. Bronchoscopy and BAL were performed in 27 young large-scale pig farmers, who had never smoked. Fifty three lifetime nonsmoking healthy students participated as controls. All farmers and controls had normal lung function (forced expiratory volume in one second (FEV1) 109 and 105% predicted respectively. Estimation of macroscopic signs of inflammation in the bronchi (erythema, oedema, secretion and friability) showed that pig farmers had significantly increased signs of inflammation. The median score was 3 (range 0-6) compared to a median score of 0 (range 0-3) in controls. More pig farmers than controls (41 versus 25%) had a positive histamine challenge (provocative dose producing a 20% fall in FEV1 (PC20) < or = 32 mg.mL-1) but the difference was not significant. The cell concentration in BAL fluid was identical in the two groups. Pig farmers had a significantly increased percentage of lymphocytes (median 7, range 1-27 versus median 2, range 0-7) and neutrophils (median 2, range 0-30 versus median 1, range 0-4) compared to controls. Spontaneous migration (19.8 versus 5.5 microns) and chemotaxis (62.6 versus 11.2 microns) was significantly increased in pig farmers compared to controls. After stimulation with zymosan and phorbol myristrate acetate (PMA), the reactive oxygen radical generation of purified alveolar macrophages was also significantly increased in pig farmers. Lifetime nonsmoking pig farmers with normal lung function have macroscopic signs of bronchial inflammation and an increased number of neutrophils in bronchoalveolar lavage. Their alveolar macrophages showed biological signs of activation. The inflammation of pig farmers bronchi may be early signs of bronchitis. PMID- 8730015 TI - The effects of recombinant human DNase on neutrophil elastase activity and interleukin-8 levels in the sputum of patients with cystic fibrosis. AB - In cystic fibrosis (CF), neutrophil-dominated airway inflammation results in high levels of neutrophil elastase (NE). Some of these proteases are sequestered by the large amounts of deoxyribonucleic acid (DNA) present in purulent sputum. Recombinant human deoxyribonuclease (rhDNase), a new treatment in CF, depolymerizes DNA. Our concerns were that this might release proteases bound to DNA, which could be potentially harmful. The in vitro and in vivo effects of rhDNase on NE and interleukin-8 (IL-8) were evaluated. The acute effects of rhDNase were evaluated in CF patients during the first 6 days of treatment. Medium-term effects were evaluated in stable CF patients observed in rhDNase over 6 months. Sputum samples were collected at regular intervals and NE activity was measured by a fluorimetric assay and IL-8 with a radioimmunoassay. In vitro addition of rhDNase resulted in a twofold increase in protease activity and this was reflected in an acute transient rise on initiation of treatment with rhDNase. Medium-term treatment was associated with a decline in NE activity and IL-8. These in vivo results are encouraging, since the increase in protease activity was transient and the trend over 6 months was a reduction in both inflammatory markers. PMID- 8730016 TI - Immunocytochemical evidence for extra-cellular initiation of elastase-induced bronchial secretory cell metaplasia in hamsters. AB - The bronchus is the only region of the hamster conducting airways to develop secretory cell metaplasia after an intratracheal instillation of human neutrophil elastase (HNE). We tested the hypothesis that this pathological change occurs because of cellular uptake of the enzyme that is specific to this region. HNE, dissolved in saline, was instilled into the trachea of hamsters, that were sacrificed 5, 15, 30 or 60 min later for immunocytochemical localization of the enzyme. Saline-treated animals served as controls. By light microscopy, HNE was evident only in the lumen and upon the epithelial surface in all airways, at all time points. Saline control tissues were negative. Electron microscopic immunogold staining revealed HNE within luminal macrophages and associated with mucus and, to a limited extent, upon the apical cell surface both in trachea and bronchus. A small amount of HNE staining occurred in the intercellular space and lamina propria of bronchi. Cytoplasmic gold particles were sparse both in treated and control animals. We conclude that instilled neutrophil elastase is excluded from the epithelial cytoplasm regardless of region. We thus reject the hypothesis of airway cellular uptake of HNE and suggest that stimulation of bronchial secretory cells to accumulate mucin granules is initiated at the cell surface, possibly by unmasking or altering region-specific receptors involved in signal transduction pathways governing mucin granule synthesis. PMID- 8730017 TI - Nucleotide-induced mucin release from primary hamster tracheal surface epithelial cells involves the P2u purinoceptor. AB - Mucin release by airway surface epithelial cells is regulated by extracellular adenosine triphosphate (ATP) via a P2 purinoceptor-mediated mechanism. The objective of the present experiment was to examine the possible involvement of uridine triphosphate (UTP) in this purinergic signal transduction pathway. Using primary hamster tracheal surface epithelial cells, ATP and UTP were compared in their abilities: 1) to displace ATP gamma S35-binding to intact cells; 2) to accumulate inositol phosphates; and 3) to stimulate mucin release. Finally, the presence of a P2u receptor message was examined. Our results showed that: 1) UTP was much less effective than ATP in displacing ATP gamma S35-binding (median inhibitory concentrations (IC50S) 240 vs 2.9 microM); 2) UTP was more potent than ATP in accumulating inositol phosphates (100 vs 43% increase at 2mM); 3) UTP was equipotent with ATP in stimulating mucin release; 4) Northern blot analysis of messenger ribonucleic acids (mRNAs) with a mouse P2u receptor complementary deoxyribonucleic acid (cDNA) probe revealed a single specific band (2.8 kb), partial sequencing of which showed a great homology with those of human or mouse P2u receptors. We conclude that, although both ATP and UTP are equipotent in stimulating mucin release, their binding kinetics to the cell surface are quite different, suggesting the presence of a common binding domain which may be responsible for the mucin release by these nucleotides. We suggest that the P2u purinoceptor is likely to be responsible for mucin release by these nucleotides, probably via activation of phospholipase C. PMID- 8730018 TI - Modulation of human bronchial epithelial cell IIICS fibronectin mRNA in vitro. AB - Fibronectin (Fn) is an extracellular matrix glycoprotein which is involved in wound repair, including repair of injured airway epithelium. Bronchial epithelial cells (BECs) are known to produce Fn which has enhanced chemotactic activity compared to serum Fn. Alternative splicing of the Fn gene is an important mechanism by which cells regulate the production of Fn. Human BECs produce Fn which contains the EIIIA region, but the expression of IIICS region variants has not previously been reported. Our purpose was to better define the molecular characteristics of human BEC Fn by determining the expression of alternative splice variants of the IIICS region of Fn of human BECs in vitro. Human bronchial epithelial cells obtained from bronchoscopy were cultured. To examine the presence of IIICS messenger ribonucleic acid (mRNA) variants, we synthesized oligonucleotide primers complementary to the published human fibronectin complementary deoxyribonucleic acid (cDNA) sequence of the IIICS domain for use in polymerase chain reactions (PCR) with total ribonucleic acid (RNA) extracted from cultured human BECs. To examine the modulation of IIICS mRNA expression, the 428 base pair (bp) DNA fragment generated in the PCR was oligo-labelled with 32[P]-deoxycytidine triphosphate (dCTP) for use as a probe for Northern blot analysis. Human BECs were cultured in the presence and absence of transforming growth factor-beta (TGF-beta) and agents which influence cyclic adenosine monophosphate (cAMP) including isoproterenol and dibutryl cAMP (db-cAMP). Total RNA from cultures was extracted, electrophoresis performed, and Northern blots obtained. Blots were hybridized with IIICS probe, total Fn cDNA, and tubulin cDNA. It was found that human BECs in culture expressed the five known human IIICS variants. TGF-beta enhanced the expression of IIICS mRNA in a concentration and time-dependent fashion. Isoproterenol and db-cAMP both reduced the expression of IIICS mRNA and attenuated the TGF-beta induction. Changes in IIICS mRNA paralleled changes in total Fn mRNA, suggesting that these agents do not selectively modulate only the IIICS domain of Fn. We conclude that human airway epithelial cell Fn in vitro does contain mRNA for five IIICS variants, and that IIICS mRNA can be modulated by TGF-beta and agents which influence cAMP. It is unknown whether alterations in IIICS variants contribute to the functional differences previously observed between airway epithelial cell Fn and plasma derived Fn. PMID- 8730019 TI - Measurement of nitric oxide in human nasal airway. AB - The nasal output of nitric oxide (NO) is known to be high, but there have been varying reports of the exact level. We attempted to establish a quantitative measurement of nasal NO, and looked for a possible relationship with nasal resistance, at rest and during exercise. Nasal airway ventilation was performed by using an air pump at a constant flow rate, whilst the soft palate was elevated voluntarily. In a preliminary study, the flow rate for sampling was changed and concentrations of NO were measured. After determination of flow rate, rhinomanometry for nasal resistance and measurement of nasal NO by chemiluminescence were carried out before and after moderate exercise. The concentration of NO ([NO]) exhibited a hyperbolic relationship with flow rate of ventilation (V'E), indicating [NO] x V'E = net nitric oxide output (V'NO) = constant. Hence, nasal NO was expressed quantitatively as V'NO (nL.min-1). For 1 L.min-1 of ventilation, [NO] varied between 2 and 500 parts per billion (ppb) (mean = 323 ppb). The average nasal V'NO in 12 healthy male subjects was 323 +/- 91 nL.min-1 (mean +/- SD). After exercise on a treadmill (10 degrees, 5 km.h-1) for 4 min, nasal V'NO decreased to 229 +/- 63 nL.min-1. At rest, expiratory and inspiratory nasal resistance was 0.27 +/- 0.04 and 0.27 +/- 0.06 Pa.s.cm-3, respectively. After exercise, expiratory and inspiratory nasal resistance decreased to 0.17 +/- 0.04 and 0.16 +/- 0.04 Pa.s.cm-3, respectively. These data indicate that nasal NO can be measured quantitatively as V'NO and might be involved in the control of nasal resistance. PMID- 8730020 TI - Role of NO in the pulmonary artery hyporeactivity to phenylephrine in experimental biliary cirrhosis. AB - The aim of this study was to see whether increased activity of nitric oxide (NO) might account for decreased pulmonary vascular tone seen in the hyperdynamic circulation of cirrhosis. We compared the pulmonary vascular reactivity of isolated pulmonary arteries (PA) from control rats (n = 10), and rats with biliary cirrhosis (n = 10) induced by chronic bile duct ligation (4 weeks). The responses of PA rings to cumulative concentrations of phenylephrine, acetylcholine, and sodium nitroprusside were studied, and also the effects of inhibition of synthesis of NO by the L-arginine analogue, N omega-nitro-L arginine (L-NOARG) in PA rings challenged with cumulative concentrations of phenylephrine and acetylcholine. The contractile response to phenylephrine was significantly reduced in cirrhotic PA rings as compared with controls. Pretreatment with L-NOARG (10(-4) M) significantly increased the contractile response to phenylephrine in PA rings from cirrhotic rats but not in control PA rings. Furthermore, L-NOARG restored the response to phenylephrine in cirrhotic PA rings back to normal. There was no difference in the relaxation of PA rings from both groups in response to acetylcholine and sodium nitroprusside. We conclude that in vitro pulmonary artery ring hyporeactivity to phenylephrine results from increased nitric oxide production in the pulmonary circulation of cirrhotic rats and might account for the hepatopulmonary syndrome. PMID- 8730021 TI - Surfactant protein-A levels increase during Pneumocystis carinii pneumonia in the rat. AB - In bronchoalveolar lavage (BAL) of human immunodeficiency virus (HIV)-infected patients with Pneumocystis carinii pneumonia and in lungs of glucocorticoid immunosuppressed rats infected with P. carinii, surfactant phospholipid levels are reduced. However, levels of the surfactant-associated protein-A (SP-A) in BAL are 4-5 times higher than normal in patients with P. carinii pneumonia. In this study, we examined the effects of glucocorticoid immunosuppression and P. carinii infection on SP-A messenger ribonucleic acid (mRNA) and protein levels in rat lungs. Rats were immunosuppressed by adding dexamethasone to their drinking water and were infected with P. carinii by intratracheal instillation of the organism. SP-A was measured by enzyme-linked immunosorbent assay (ELISA) and SP-A mRNA by hybridization of Northern blots with an SP-A complementary deoxyribonucleic acid (cDNA) probe. There was a severalfold increase in SP-A protein and mRNA levels in uninfected glucocorticoid-treated rats. However, contrary to what has been reported with the surfactant-associated lipids, SP-A mRNA and protein levels in P. carinii-infected animals were significantly higher than those found in the uninfected, immunosuppressed animals. Our results demonstrate that SP-A increases, probably as a result of elevated mRNA levels, in immunosuppressed rats with P. carinii infection and are consistent with our findings in HIV-positive patients with P. carinii pneumonia. PMID- 8730022 TI - Long-term canine exposure studies with ambient air pollutants. AB - Dogs are often the species of choice as an experimental model for the study of pulmonary responses to long-term exposure to air pollutants in chambers simulating environmental or occupational exposure in man. Their lungs bear a reasonable resemblance to human lungs, they are large enough to allow serial measurements of pulmonary responses, and they live long enough to ensure that findings are not confounded by aging. Several long-term canine exposure studies with ambient air pollutants have been performed since 1957: seven studies with gaseous and particulate sulphur (IV); three studies with nitrogen oxides; three studies with ozone; two studies with acidic particles; three studies with mixtures of sulphurous pollutants that might have resembled the 1952 London smog; and one study in which raw and ultra violet (UV)-irradiated motor vehicle exhaust and sulphurous pollutants were used. The findings support the hypothesis that long-term exposure to air pollutants at ambient levels might cause bronchitic lesions (sulphur oxide), emphysematous lesions (nitrogen dioxide) or fibrotic lesions (ozone). None of the studies showed an indication of synergistic effects. To improve our understanding of pulmonary responses initiated by the inhalation of pollutants over long periods of time, new concepts are needed. Investigators should consider studies with canine models of cardiopulmonary diseases, the application of novel immunological and molecular biology techniques, the phenomena of tolerance and adaptation to inhaled air pollution, and exposure atmospheres with increasing complexity, including fine and ultrafine particles. PMID- 8730023 TI - Bronchodilator delivery with metered-dose inhalers in mechanically-ventilated patients. AB - Metered-dose inhalers (MDIs) provide several advantages over nebulizers, including ease of administration, decreased cost, reliability of dosing, and freedom from contamination. However, this method of aerosol delivery has been considered ineffective in mechanically-ventilated patients because most of the aerosol deposits in the endotracheal tube and ventilator circuit. A smaller amount of aerosol from a MDI is deposited in the lower respiratory tract of mechanically-ventilated patients than in ambulatory patients, although recent studies show that a significant bronchodilator effect can still be achieved. When employed optimally, significant bronchodilation occurs with as little as 4 puffs of a sympathomimetic aerosol. Multiple factors influence the efficacy of MDIs in mechanically-ventilated patients. The method of connecting the MDI canister to the ventilator circuit has a marked effect on aerosol delivery, and other factors include the timing of MDI actuation, ventilator mode, tidal volume, circuit humidity, and duty cycle. With a proper technique of administration, a MDI serves as an effective, convenient, and safe method for delivering bronchodilator aerosols in mechanically-ventilated patients. PMID- 8730024 TI - Colony-stimulating factors as an adjunct to chemotherapy in small cell lung cancer. AB - Myelosuppression is the major dose-limiting toxicity of chemotherapy in small cell lung cancer (SCLC). The capacity of colony stimulating factors (CSFs) to stimulate granular neutrophil recovery may be of great value to prevent or cure febrile neutropenia and to increase dose-intensity. The aim of this review was to assess the current use of CSFs in SCLC on the basis of experimental and clinical data. Primary CSF administration has been shown to reduce the incidence of febrile neutropenia, hospital admission rate, and antibiotic use subsequent to cyclophosphamidedoxorubicin-high dose etoposide (CDE) chemotherapy, without improvement of survival or disease control. Primary CSF administration may be recommended when the expected incidence of febrile neutropenia is at least 40%. This benefit has not been established with less myelosuppressive regimens, such as cisplatin-etoposide (PE), which remains an alternative combination of SCLC when standard doses are used. A trial comparing high-dose CDE + CSF with PE would be of considerable interest. There is currently little clinical basis for the use of CSFs to increase chemotherapy dose-intensity, outside clinical trials. Peripheral blood progenitor cells mobilized with CSFs offer interesting prospects. Further studies, with later initiation, shorter duration or lower doses of CSFs, are warranted to improve the cost-effectiveness of CSFs. CSF therapy in addition to antibiotics is normally not justified in febrile neutropenia, except perhaps in selected patients with sepsis syndromes, hypotension or pneumonia. PMID- 8730025 TI - Preparation of bronchoalveolar lavage fluid with microscope slide smears. AB - The method of preparation of bronchoalveolar lavage fluid (BALF) for cytological examination can significantly affect the results of cellular quantitation. Investigations have shown that cytocentrifugation leads to an underestimation of the number of lymphocytes and membrane filter preparation to an underestimation of the number of neutrophils. As a simple alternative to these two techniques, BALF cells could be prepared by the microscope slide smear technique, which is familiar as the means for preparing peripheral blood for differential counts. In order to compare cell differentials determined by microscope slide technique with differentials resulting from cytocentrifugation, cells were isolated from 35 BALF samples using standard methods, and counted using a haematocytometer. Forty thousand cells in 200 microL were prepared by cytocentrifugation (3 min, 57 x g; Cytospin 2) and 5 x 10(5) cells in 5 microL by microscope slide smear. Both samples were air-dried, stained using May-Grunwald Giemsa stain, and 600 cells were counted to obtain differentials. To test the adequacy of sampling by the microscope slide smear technique, known quantities of lymphocytes or neutrophils were added to fixed numbers of BALF cells, microscope slide smears prepared, and differentials determined on 600 cells. The resulting differentials were compared to the calculated differentials. Preparation of BALF cells with the microscope slide smear technique yielded well-preserved cell morphology. Compared to cytocentrifugation, microscope slide smear preparations had significantly higher percentages of lymphocytes. The microscope slide smears for the samples with predetermined numbers of cells yielded lymphocyte and neutrophil percentages which did not differ from the calculated differentials (59.6 +/- 1.5 vs 59.6 +/- 5.2% and 54.6 +/- 6.0 vs 53.1 +/- 6.0%, respectively). Varying the number of cells counted from 100 to 800 confirmed the reproducibility of the counts for counting 600 cells. Using 5 x 10(5), 2.5 x 10(5), or 1 x 10(5) cells per preparation demonstrated that adequate specimens could be obtained from as few as 1 x 10(5) cells. Thus, microscope slide smear preparation is a simple and accurate method for the quantitation of bronchoalveolar lavage fluid cytology. PMID- 8730026 TI - Stent flexibility: an essential feature in the treatment of dynamic airway collapse. AB - Implantation of endobronchial stents for treatment of dynamic airway collapse represents a suitable therapeutic option to alleviate distressing symptoms. We report the case of a 43 year old patient suffering from progressive respiratory distress 2 weeks after insertion of a balloon-expandable radial noncompliant Palmaz stent in an unstable segment of the left main bronchus, with the aim of preventing symptomatic airway collapse. Bronchial instability had developed following sleeve resection of the right lung due to adenoid cystic carcinoma. Explanation revealed compression and deformation of the stent. Peak expiratory flow (PEF) had declined a low of 1.38 L.s-1 (forced expiratory volume in one second (FEV1) 1.02 L). With placement of a Strecker stent, having the ability to re-expand within certain limits, bronchial collapse could be avoided and marked clinical improvement as well as expiratory flow increase was noted (PEF 7.10 L.s 1; FEV1 = 2.03 L). At 13 months follow-up, clinical status was unchanged. A decline in forced expiratory flow (PEF 5.96 L.s-1; FEV1 1.69 L), however, indicated a possible change in the structural integrity of the Strecker stent. We conclude that physical properties of endobronchial stents may be crucial for good functional results in major airway collapse. Stiff prostheses, when compressed, can induce severe airway obstruction. PMID- 8730027 TI - Plastic bronchitis. AB - Plastic bronchitis is generally associated with some type of pulmonary disease and improves either spontaneously or with medical therapy. We present a case of plastic bronchitis with no known cause. The patients' symptoms were not responsive to medical therapy but were relieved after right middle lobectomy. PMID- 8730028 TI - Fluoxetin-induced pulmonary granulomatosis. AB - A patient treated with fluoxetin for a manic depressive disorder developed pulmonary inflammatory nodules with noncaseating giant cell granulomas, interstitial pneumonia and non-necrotizing vasculitis, whilst remaining asymptomatic. A progressive resolution of pulmonary nodules occurred after withdrawal of the offending agent, and the chest radiograph returned to normal in 9 months. The diagnosis was assessed by an open lung biopsy. PMID- 8730029 TI - A young man with fever, dyspnoea and nonproductive cough. PMID- 8730030 TI - Neurophysiology of the cough reflex. PMID- 8730031 TI - Harvey Fletcher's role in the creation of communication acoustics. AB - As the reader might appreciate after reading Fletcher's 1953 views, in 1918 Fletcher had taken on the toughest problem of all: to quantify and model how we hear and understand speech. This understanding allowed AT&T Bell Labs engineers to develop the necessary specifications of what was to become the largest telephone network in the world. The problems that Fletcher and his colleagues studied were so complicated, and took so many years, that it has been difficult to appreciate the magnitude of their accomplishments. It is therefore understandable why his work has had such a great impact on our lives. Almost single-handedly he created the fields of communication acoustics and speech and hearing as we know them today. Everyone who has ever used the telephone has reaped the benefit provided by this man and his genius. von Bekesy, Davis, Stevens, and Zwicker are some of the names that come to mind when we think of hearing. Bell invented the telephone, and Edison made it into a practical device. Harvey Fletcher may not be as well known as these men today, but his scientific contributions to the fields of telephony, hearing, and human communication are absolutely unsurpassed. Given this present opportunity to reflect back on this great man, I would describe Harvey Fletcher as the singular intellectual force in the development of present-day communication acoustics and telephony. PMID- 8730068 TI - Fractal noise strength in auditory-nerve fiber recordings. AB - Discharge patterns recorded from single auditory-nerve fibers have demonstrated long-range dependence, with the count variance-to-mean ratio growing as a power of the counting time for times greater than 0.1-1 s. The intent of this study is to provide a large dataset to enable a more detailed investigation of this phenomenon. Based on 108 recordings from a cat, we conclude that the presence of the fractal noise in the discharge rate is independent of characteristic frequency and stimulus level, but does depend on discharge rate. We measured the low-frequency power of the fractal noise, finding its coefficient of variation to range between 6% and 26% and to decrease as firing rate increases. Such behavior is consistent with multiplicative fractal variations in models of the hair cell membrane permeability to neurotransmitter. Measured standard deviations of spike rate correspond to a sound-pressure level difference limen of approximately 1 dB. PMID- 8730069 TI - A functional model of the hair cell-primary fiber complex. AB - A model is proposed containing three parallel systems; system 1 is a single channel consisting of an inhibitory circuit being fed by a fixed permeability responsible for spontaneous activity in parallel with a displacement-driven permeability, while systems 2 and 3 each consist of a number of identical permeability-volume-permeability channels with on/off switches activated at levels specific for each channel. The (continuous) output of each system drives a Poisson generator whose output "events" are delivered to a firing mechanism in the form of a "leaky integrator" delivering actual "firings" in simulated real time. The model quite accurately reproduces (1) stimulation and recovery phases of PST histograms obtained at six levels of stimulation for six auditory fibers (including "dead" period after stimulation), (2) single-exponential functions of recovery from adaptation for both onset and steady-state part of probe tone, (3) increments in response as independent of time of application of stimulus increment, (4) a specified hazard function for firings, (5) interval histograms of driven activity, (6) loss of phase locking with increasing frequency of stimulation, and (7) extended dynamic range for onset of response. The model fails in reproducing specific characteristics for response to decrements, and for response to AM. PMID- 8730070 TI - Linearity of sound transmission through the human skull in vivo. AB - The linearity of sound propagation through the human skull was investigated. One male subject, equipped with bilateral skin-penetrating titanium fixtures for attachment of bone-anchored hearing aids, was studied thoroughly. Three different methods were used: comparison of the frequency response functions estimated at different signal levels (using stepped sine as well as random noise), comparison of the coherence function at different signal levels (using random noise), and the Hilbert transform of the estimated frequency response function. Frequencies from 0.1 to 10 kHz and signal levels up to 77 dB HL at discrete frequencies were used. No indication of any significant nonlinear behavior was found with the three methods used. PMID- 8730071 TI - Biophysics of the cochlea. II: Stationary nonlinear phenomenology. AB - Nonlinearities affecting cochlear mechanics produce appreciable compression in the basilar membrane (BM) input/output (I/O) functions at the characteristic frequency for sound-pressure levels (SPLs) as low as 20 dB (re: 20 microPa). This is thought to depend upon saturation of the outer hair cell (OHC) mechanoelectrical transducer (MET). This hypothesis was tested by solving a nonlinear integrodifferential equation that describes the BM vibration in an active cochlea. The equation extends a previously developed linear approach [Mammano and Nobili, J. Acoust. Soc. Am. 93, 3320-3332 (1993)], here modified to include saturating MET, with a few corrections mainly concerning tectorial membrane resonance and OHC coupling to the BM. Stationary solutions were computed by iteration in the frequency domain for a wide range of input SPLs, generating BM I/O functions, frequency response envelopes, and two-tone distortion products. Traveling-wave amplitude envelopes were also computed for a fixed suppressor and several suppressed tones in order to evidence the phenomenon of two-tone suppression (frequency masking) at the mechanical level. All results accord nicely with experimental data. PMID- 8730072 TI - Vibrotactile threshold in young and old observers: the effects of spatial summation and the presence of a rigid surround. AB - Human psychophysical detection thresholds for ten frequencies of sinusoidal vibration ranging from 10 to 400 Hz were obtained on the left index fingertip and thenar eminence of young and older observers using a three-alternative forced choice tracking procedure. The first experiment utilized a 7-mm (0.38 cm2) contactor and rigid surround with 1-mm gap. In the second experiment, three contactor sizes (1.6-, 7.0-, and 25.4-mm diameter) and two surround configurations (1-mm gap between contactor and surround, and no surround) were used. The results indicate that, although the shapes of the threshold versus frequency functions were similar in the two age groups, there was a reduction in sensitivity for the older group at all frequencies. Furthermore, taking into account the difference in sensitivity between the two age groups, spatial summation and the effects of a surround did not seem to differ between the two groups. These results are discussed in the context of physiological models of cutaneous sensitivity and changes in receptor function with age. PMID- 8730073 TI - Computer simulation of auditory stream segregation in alternating-tone sequences. AB - A computer model is described that takes a novel approach to the problem of accounting for perceptual coherence in alternating pure-tone sequences by using simple physiological principles that operate at a low level. Using the same set of parameter values, the model is able to reproduce a number of phenomena associated with auditory stream segregation. These are (1) the buildup of stream segregation over time, (2) the temporal coherence and fission boundaries obtained from human listeners, and (3) the trill threshold. Whereas these phenomena are generally accounted for in terms of an auditory scene-analysis process that works on the basis of Gestalt perceptual principles, the operation of the model suggests that some Gestalt auditory grouping may be the product of low-level processes. PMID- 8730074 TI - The effect of test signal type and bandwidth on the categorical scaling of loudness. AB - Recently several methods for obtaining clinical measures of loudness growth through the use of categorical scaling (CS) have been proposed for the selection of hearing aids. These methods use differing test signals or suggest frequency specific level corrections in an attempt to reflect the loudness perception of hearing aid-processed speech. While some decisions regarding the stimuli utilized for loudness perception procedures are based on measured relationships to speech signals, the effect on loudness perception of changing signal type and bandwidth (as measured by CS) remains unclear. The relationships between the CS loudness growth of signals of differing type, (pure tones, noise bands, filtered/temporally inverted/passband speech) and bandwidth were examined for subjects with both normal and impaired hearing. Results suggest that when the bandwidth is similar (e.g., pure tone and 1/3 oct), signal type does not have significant bearing on loudness perception. As expected, increasing the bandwidth beyond the critical band affected loudness growth, as wideband speech stimuli were judged to be significantly louder than narrow-band speech at equivalent overall sound levels. In this investigation, similar loudness growth patterns were noted across category ratings for all test signal bandwidths. In contrast, loudness growth data obtained using an intelligible speech signal [Cox et al., The American Academy of Audiology (1994a)], revealed loudness growth patterns for speech were fundamentally different than those obtained for tones. Implications for hearing aid fitting strategies are discussed. PMID- 8730075 TI - Stimulus-driven, time-varying weights for comodulation masking release. AB - This study tests the hypothesis that comodulation masking release (CMR) is mediated by "listening in the valleys" [S. Buus, J. Acoust. Soc. Am. 78, 1958 1965 (1985)]. Detectability was measured for signals consisting of six consecutive 25-ms, 1-kHz tone pulses presented in a 50-Hz-wide masker or in maskers consisting of seven 50-Hz-wide noises, one critical band apart, with either correlated or uncorrelated envelopes. The level of each signal pulse varied randomly around masked threshold according to Gaussian distributions with rms perturbations (standard deviations) of 3 or 6 dB. For each listener and condition, the responses from 5000 trials were sorted to construct conditional psychometric functions for d' as a function of signal-pulse intensity for ten ranges of short-term level of the on-frequency masker band during the pulse. The slopes of these functions for three normal listeners decrease markedly with increasing short-term masker level for the correlated multiband masker, but are largely constant for the other maskers. This indicates that the weight applied to the signal channel is high when the masker level is low and vice versa for the correlated masker, but is approximately constant for single-band and uncorrelated multiband maskers. These findings provide direct evidence that CMR is mediated by "listening in the valleys," but models based on direct envelope comparison may also account for the results if they are modified to include a compressive nonlinearity before the comparison. PMID- 8730076 TI - Decision rules of listeners in spectral-shape discrimination with or without signal-frequency uncertainty. AB - A correlation technique was used to assess the decision rules of three listeners in two cases of spectral-shape discrimination tasks. In one case the signal frequency was fixed, and in the other it was randomly varied within each block of trials. In order to estimate the decision rule of the listeners, the experimenter superimposed random level perturbations on each frequency component upon each stimulus presentation. Over many trials, correlation coefficients were computed between the random perturbations and the binary responses of the listeners, and were expressed as a function of signal level. For both the fixed- and random signal cases, the measured correlation functions were in reasonably good agreement with those predicted based on the likelihood-ratio decision rules. Thus the listeners appeared to use information nearly optimally in discriminating spectral shapes. This investigation demonstrated that the correlation technique can be used to reveal the decision rules for cases where the decision statistics are nonlinear functions of the observations. PMID- 8730077 TI - Identification of multidimensional complex sounds having parallel dimension structure. AB - The present series of experiments examined the ability of normal-hearing listeners to make use of cues from multiple, independent stimulus dimensions when classifying multidimensional complex sounds. Ten listeners classified complex sound pulses that differed along three independent dimensions. The stimuli were 100 ms in duration and were synthesized using five simultaneous sinusoids. The three dimensions of the complex stimuli manipulated in this experiment were harmonicity, spectral shape, and amplitude envelope. Each stimulus dimension could take on one of two values, referred to here as target or nontarget. Eight stimuli were synthesized using all possible combinations of the dimension values. Subjects were trained to label two stimuli, the two having either all-target or all-nontarget values, as "+" and "0," respectively. Following this training, subjects were asked to classify all eight stimuli as either "+" or "0." Results of this experiment indicated that listeners preferred to classify these stimuli on the basis of one stimulus dimension. However, the preferred dimension was not the same for all of the listeners. In addition, it was demonstrated that it was possible to train an individual to use a dimension other than the one or two initially preferred when classifying the stimuli. PMID- 8730078 TI - Temporal asymmetry in the auditory system. AB - When a damped exponential with a half-life of 4-8 ms is repeated every 25-50 ms and used to modulate a sinusoid or a wideband noise, it suppresses the sound quality typically associated with the carrier. When the envelopes of these "damped" sounds are reversed in time, producing "ramped" sounds, a continuous component with the sound quality of the carrier is restored to the perception. This paper presents an experiment that measures the temporal asymmetry revealed by this perceptual contrast. A ramped sinusoid or noise with a given half-life was presented with a damped sinusoid or noise having the same or greater half life, to determine the damped half-life required to produce a continuous component with the equivalent relative strength in the two sounds. The results with sinusoidal carriers show that the half-life of the damped sound has to be, on average, about five times the half-life of the ramped sound if the tonal component of the two perceptions is to have the same relative strength. The asymmetry for the noise carrier is about half that of the sinusoidal carrier and, again, the damped sound has the greater matching half-life. Several multichannel auditory models based on a gammatone filterbank are used to try to explain the data in terms of traditional leaky integration, but they produce neither sufficient asymmetry nor the correct pattern of asymmetry. A "delta-gamma" theory is then developed to provide a framework for understanding temporal asymmetry in the auditory system. The theory is used to compare the temporal asymmetry produced by several auditory models and to explain when and how they can accommodate the perceptual asymmetry observed in the experiments. PMID- 8730079 TI - Vowel identification based on amplitude modulation. AB - This study investigated the extent to which flat-spectrum harmonic complexes could be identified as one of six vowels when three pairs of successive harmonics, located at the first, second, and third formant frequency values, were amplitude modulated. In experiment 1, the amplitude modulation (AM) rate was at or close to 10 Hz. In condition 1, all components were added in cosine phase, and the 10-Hz AM was in phase for all "formants." Performance improved monotonically with increasing modulation index, m. In condition 2, m was fixed at 0.5 and the level of each background harmonic was varied randomly (roved) from stimulus to stimulus. Even a rove range of only +/- 2 dB reduced scores considerably. Condition 3 was like condition 1, but with components added in random phase. Performance was very poor for all modulation indices. This suggests that subjects were unable to use momentary differences in level between formant and background harmonics, and supports the idea that, for cosine-phase stimuli, they were using information from the low-amplitude portions ("valleys") of the cochlea-filtered waveforms. In further conditions, the components were added in cosine phase and the AM had a different phase and/or different rate (10, 16, and 24 Hz) on the different formants. Scores were very similar to those obtained when the AM was identical for all formants. In experiment 2, the AM rate was at or close to 2 Hz. When all formants were modulated in phase at 2 Hz, very good performance was found for components added in cosine phase, and performance was essentially unaffected by making the AM different in rate and/or phase across formants. When the components were added in random phase, performance was well above chance when the formants were modulated in-phase at 2-Hz, but worsened markedly when the modulation differed in rate and/or phase across formants. Randomizing the level of each background harmonic caused performance to deteriorate and to become similar for cosine-phase and random-phase stimuli. Performance deteriorated further when the AM differed in phase across formants. The results suggest that, for a 2-Hz modulation rate, and when information from the valleys is not available, performance depends on momentary increases in level of the formant harmonics relative to the background. PMID- 8730080 TI - A longitudinal investigation of duration and temporal variability in children's speech production. AB - A number of cross-sectional, acoustic studies have found that young children's speech segments tend to be longer and more variable than those of older children and adults. However, very little longitudinal information of this nature is available that considers changes across time for individual children. The present investigation is a longitudinal analysis of several temporal characteristics of the speech of 12 children of various ages who were each seen twice, approximately 1 1/2 years apart. For the group, durations decreased on average from the initial to the follow-up recordings by approximately 10%, and temporal variability decreased by about 40%. For the individual children, however, it was found that some of them showed few, if any, changes in some of the temporal measurements made at the two different times, whereas others showed substantial differences. Younger children also did not necessarily show longer durations or greater variability than older children, nor did younger children always show greater changes across time than older children. Thus, although cross-sectional studies indicate that there is a general tendency when comparing groups for increased age to be associated with shorter durations and reduced variability, individual children may not evidence such patterns or changes across time. PMID- 8730081 TI - Perception of back vowels: effects of varying F1 - F0 Bark distance. AB - In a study of vowel height perception using front vowels, Hoemeke and Diehl [J. Acoust. Soc. Am. 96, 661-674 (1994)] found that F1 - F0 distance was the best predictor of perceived vowel height for the phonological distinction [+/- high], while for two other vowel height distinctions F1 alone was the best predictor. Further, the [+/- high] identification function was defined by a sharp boundary located at 3- to 3.5-Bark F1 - F0 distance. One hypothesis offered was that F1 - F0 distance had cue value for the [+/- high] distinction because of an underlying quantal region on the F1 - F0 distance dimension. However, the results are also predicted if it is supposed that F1 - F0 distance is a cue for vowel height only for pure height distinctions. The present study further tested these possibilities, using back vowels. The results allowed us to reject both as general explanations of vowel height perception. However, the results were consistent with a third possible explanation, namely, that phonetic quality is determined by the tonotopic distances between any adjacent spectral peaks (e.g., F3 - F2, F2 - F1, and F1 - F0), with greater perceptual weight accorded to smaller distances. PMID- 8730082 TI - Effect of temporal modulation reduction on spectral contrasts in speech. AB - In this paper the effect of temporal modulation reduction on spectral contrasts is investigated. First, a spectral modulation transfer function (SMTF) is presented as a method to measure the transfer of spectral ripples (sinusoidal periods/oct) in the short-time spectral envelope by comparing the spectral modulation depth of original and processed speech fragments. Measuring the SMTF for speech subjected to uniform reduction of the temporal modulation depth (i.e., modulation-frequency-independent reduction) in 24 1/4-oct bands showed an almost equal uniform reduction of the spectral modulations. Furthermore, the SMTF was used to measure the reduction of spectral contrasts associated with low-pass and high-pass temporal-envelope filtering [Drullman et al., J. Acoust. Soc. Am.95, 1053-1064 and 2670-2680 (1994a, b)]. For a perceptual evaluation, sentences were processed to reduce spectral contrasts and the speech-reception threshold (SRT) in noise was measured with ten normal-hearing subjects. Comparison of the results with those obtained previously after temporal-envelope filtering revealed that the SRT-effect of temporal high-pass filtering can be completely accounted for by the associated reduction of spectral contrasts. However, this relationship cannot be demonstrated conclusively in the case of temporal low-pass filtering. PMID- 8730083 TI - Quad-band excitation for low bit rate speech coding. AB - The excitation parameters of a multiband excitation model for speech production consist of a pitch period and a series of voiced/unvoiced (v/uv) decisions for a number of fixed frequency bands around the harmonics of the fundamental. In this paper, it is shown that, at most, four v/uv variable-length nonoverlapping frequency bands spanning over the telephone bandwidth are adequate to represent the excitation spectrum that provides an improved modeling of mixed voicing and noisy speech. A quad-band excitation (QBE) method is presented which generates excitation source for a linear prediction vocoder for low bit rate transmission of speech signal and the DRT scores validate the potential of the method. PMID- 8730084 TI - Wind-instrument reflection function measurements in the time domain. AB - Theoretical and computational analyses of wind-instrument sound production in the time domain have emerged as useful tools for understanding musical instrument acoustics, yet there exist few experimental measurements of the air-column response directly in the time domain. A new experimental, time-domain technique is proposed to measure the reflection function response of woodwind and brass instrument air columns. This response is defined at the location of sound regeneration in the mouthpiece or double reed. A probe assembly comprised of an acoustic source and microphone is inserted directly into the air column entryway using a foam plug to ensure a leak-free fit. An initial calibration phase involves measurements on a single cylindrical tube of known dimensions. Measurements are presented on an alto saxophone and euphonium. The technique has promise for testing any musical instrument air columns using a single probe assembly and foam plugs over a range of diameters typical of air-column entryways. PMID- 8730085 TI - Acoustic nonlinearity parameter tomography for biological specimens via measurements of the second harmonics. AB - The acoustic nonlinearity parameter B/A is a new parameter in ultrasound tissue characterization. In this paper, on the basis of a theoretical analysis of finite amplitude ultrasonic wave propagation in fluidlike media, an experimental system for nonlinearity parameter tomography is developed. Using this system, the amplitude of the second harmonic wave cumulated in the propagation path of the primary wave is measured and the finite amplitude insert-substitution method is used in the conventional computer tomography (CT) technique. The filtered convolution algorithm is implemented using the amplitude of the second harmonic as the projection data, then the B/A tomography can be reconstructed. The B/A images for several biological samples are obtained. Besides, the image of porcine pathological liver tissue is studied and is compared with that of porcine normal liver tissue. These results show the potential applications of the nonlinearity parameter B/A in medical diagnosis. PMID- 8730086 TI - Effect of static pressure on acoustic transmittance of Albunex microbubble suspensions. AB - Albunex (ALX), an albumin-stabilized microbubble echo contrast agent, is sensitive to pressures similar to those produced by the heart. The tested hypothesis was that the acoustic transmittance of ALX suspensions will increase with increasing hydrostatic pressure (Ps). The test involved an acoustic setup analogous to a spectrophotometer. The acoustic transmittance of microbubble suspensions was strongly Ps dependent. Transmittance at 1 MHz was essentially zero at ambient pressure, increasing to approximately 50%, approximately 63%, and nearly 100% at Ps of 80, 120, and 400 mm Hg, respectively. The ultrasound pulses used to interrogate samples were without measurable effect on the acoustic transmittance of suspensions maintained at ambient pressure during experimental measurements. The data indicate that many of the microbubbles are destroyed at Ps comparable to those produced by the heart. PMID- 8730087 TI - Acoustic basis for recognition of aspect-dependent three-dimensional targets by an echolocating bottlenose dolphin. AB - The relationships between acoustic features of target echoes and the cognitive representations of the target formed by an echolocating dolphin will influence the ease with which the dolphin can recognize a target. A blindfolded Atlantic bottlenose dolphin (Tursiops truncatus) learned to match aspect-dependent three dimensional targets (such as a cube) at haphazard orientations, although with some difficulty. This task may have been difficult because aspect-dependent targets produce different echoes at different orientations, which required the dolphin to have some capability for object constancy across changes in echo characteristics. Significant target-related differences in echo amplitude, rms bandwidth, and distributions of interhighlight intervals were observed among echoes collected when the dolphin was performing the task. Targets could be classified using a combination of energy flux density and rms bandwidth by a linear discriminant analysis and a nearest centroid classifier. Neither statistical model could classify targets without amplitude information, but the highest accuracy required spectral information as well. This suggests that the dolphin recognized the targets using a multidimensional representation containing amplitude and spectral information and that dolphins can form stable representations of targets regardless of orientation based on varying sensory properties. PMID- 8730088 TI - Some acoustical properties of the otic bones of a fin whale. AB - The otic bones in this report are the tympanic bulla, the periotic, and the three ossicles (malleus, incus, and stapes) of an adult fin whale (Balaenoptera physalus). The purpose was to determine if the periotic was denser than the other otic bones. It was found in one male adult fin whale that the density of all the otic bones is approximately the same, 2.50 kg/m3 with a maximum of 2.58. The lowest density was observed in the stapes (2.36). The sonic velocity seems to vary as the density but there also seems to be a structural effect. The maximum sonic velocity was 4.89 km/s in the malleus. The specific acoustic impedance was as high as 12.5 megarayles in the periotic. These values compare with those for human femur of 1.95 for the density, 3.73 for the sonic velocity, and 7.33 for the specific acoustic impedance. The ossicles weigh as much as 200 times as much as human ossicles. The density of whale ossicles are about ten percent greater than human ossicles. The mechanical natural frequency of the whale ossicles must be very low. The approximate uniformity of the properties of this whale's otic bones may be characteristic of the middle ear. The density of the otic bones of land mammals is less than for whales. The density of the horse petrosal (2.29 g/cc) is essentially the same as the density of adult human ossicles (2.23-2.27 g/cc). The high density of the otic bones for all mammals suggests it may be related to hearing acuity perhaps by increasing the specific acoustic impedance, which increases the acoustic contrast with the other body tissues. PMID- 8730094 TI - The tyrosine kinase receptors Ron and Sea control "scattering" and morphogenesis of liver progenitor cells in vitro. AB - The mammalian RON and the avian sea genes encode tyrosine kinase receptors of poorly characterized biological functions. We recently identified macrophage stimulating protein as the ligand for Ron; no ligand has yet been found for Sea. In this work we investigated the biological response to macrophage-stimulating protein in mouse liver progenitor cells expressing Ron. These cells were also transfected with a chimeric cDNA encoding the cytoplasmic domain of Sea, fused to the extracellular domain of Trk (nerve growth factor receptor). In the presence of nanomolar concentrations of the respective ligands, both receptors induced cell "scattering", extracellular matrix invasion, and DNA synthesis. When liver progenitor cells were grown in a tri-dimensional type-I collagen matrix, ligand induced stimulation of either Ron or Sea induced sprouting of branched cell cords, evolving into ductular-like tubules. The motogenic, mitogenic, and morphogenic responses were also elicited by triggering the structurally related hepatocyte growth factor receptor (Met) but not epidermal growth factor or platelet-derived growth factor receptors. These data show that Ron, Sea, and Met belong to a receptor subfamily that elicits a distinctive biological response in epithelial cells. PMID- 8730095 TI - Drosophila Cdk8, a kinase partner of cyclin C that interacts with the large subunit of RNA polymerase II. AB - A number of cyclins have been described, most of which act together with their catalytic partners, the cyclin-dependent kinases (Cdks), to regulate events in the eukaryotic cell cycle. Cyclin C was originally identified by a genetic screen for human and Drosophila cDNAs that complement a triple knock-out of the CLN genes in Saccharomyces cerevisiae. Unlike other cyclins identified in this complementation screen, there has been no evidence that cyclin C has a cell-cycle role in the cognate organism. Here we report that cyclin C is a nuclear protein present in a multiprotein complex. It interacts both in vitro and in vivo with Cdk8, a novel protein-kinase of the Cdk family, structurally related to the yeast Srb10 kinase. We also show that Cdk8 can interact in vivo with the large subunit of RNA polymerase II and that a kinase activity that phosphorylates the RNA polymerase II large subunit is present in Cdk8 immunoprecipitates. Based on these observations and sequence similarity to the kinase/cyclin pair Srb10/Srb11 in S. cerevisiae, we suggest that cyclin C and Cdk8 control RNA polymerase II function. PMID- 8730096 TI - Cell adhesion-dependent inactivation of a soluble protein kinase during fertilization in Chlamydomonas. AB - Within seconds after the flagella of mt+ and mt- Chlamydomonas gametes adhere during fertilization, their flagellar adenylyl cyclase is activated several fold and preparation for cell fusion is initiated. Our previous studies indicated that early events in this pathway, including control of adenylyl cyclase, are regulated by phosphorylation and dephosphorylation. Here, we describe a soluble, flagellar protein kinase activity that is regulated by flagellar adhesion. A 48 kDa, soluble flagellar protein was consistently phosphorylated in an in vitro assay in flagella isolated from nonadhering mt+ and mt- gametes, but not in flagella isolated from mt+ and mt- gametes that had been adhering for 1 min. Although the 48-kDa protein was present in the flagella isolated from adhering gametes, we demonstrate that its protein kinase was inactivated by flagellar adhesion. Immunoblot analysis and inhibitor studies indicate that the 48-kDa protein in nonadhering gametes is phosphorylated by a protein tyrosine kinase. In vivo experiments showing that the protein tyrosine phosphatase inhibitor sodium orthovanadate inhibits fertilization suggest that protein dephosphorylation may be required for signal transduction. The 48-kDa protein and its protein kinase may be among the first elements of a novel signalling pathway that couples interaction of flagellar adhesion molecules to gamete activation. PMID- 8730097 TI - SDC25, a dispensable Ras guanine nucleotide exchange factor of Saccharomyces cerevisiae differs from CDC25 by its regulation. AB - The SDC25 gene of Saccharomyces cerevisiae is homologous to CDC25. Its 3' domain encodes a guanine nucleotide exchange factor (GEF) for Ras. Nevertheless, the GEF encoded by CDC24 is determinant for the Ras/cAMP pathway activation in growth. We demonstrate that the SDC25 gene product is a functional GEF for Ras: the complete SDC25 gene functionally replaces CDC25 when overexpressed or when transcribed under CDC25 transcriptional control at the CDC25 locus. Chimeric proteins between Sdc25p and Cdc25p are also functional GEFs for Ras. We also show that the two genes are differentially regulated: SDC25 is not transcribed at a detectable level in growth conditions when glucose is the carbon source. It is transcribed at the end of growth when nutrients are depleted and in cells grown on nonfermentable carbon sources. In contrast, CDC25 accumulation is slightly reduced when glucose is replaced by a nonfermentable carbon source. PMID- 8730098 TI - Contrasting effects of the SATB1 core nuclear matrix attachment region and flanking sequences of the keratin 18 gene in transgenic mice. AB - The 2.3 kb and 3.5 kb of DNA that flank the human keratin 18 (K18) gene and synthetic nuclear matrix attachment regions (MAR) composed of the binding sites for the SATB1 nuclear protein were fused to a reporter gene that utilizes the mouse metallothionein promoter and the human growth hormone gene (MThGH). Transgenic mice were generated from both constructions and the control MThGH gene to test K18 and SATB1 MAR sequences for the ability to insulate the reporter gene from integration site-specific position effects. The MThGH control gene was variably expressed in brain, heart, intestine, kidney, liver, and testes, confirming previous studies. In contrast, the MThGH gene insulated by the K18 flanking sequences was expressed in the same tissues of four independent transgenic animals at levels correlated with the copy number except for intestine. The average level of expression on a per gene basis of the K18 insulated gene was from 9- to 49-fold higher than the control. The MThGH gene linked to the SATB1 MAR sequences was completely repressed in the brains and kidneys of all six transgenic mice. However, expression was nearly as efficient in testes as the K18-insulated gene. Both the SATB1 MAR and the K18 flanking sequences confer position-independent transcriptional status on the reporter gene in some or many tissues. However, the effects are stimulatory for the K18 elements and generally suppressive for the SATB1 MAR elements. PMID- 8730099 TI - Role of cyclin A and p27 in anti-IgM induced G1 growth arrest of murine B-cell lymphomas. AB - Cross-linking surface immunoglobulin (Ig)M on the WEHI-231 B-cell lymphoma results in decreased cell size, G1/S growth arrest, and finally DNA cleavage into oligonucleosomal fragments that are the classical features of apoptotic cells. Treatment of WEHI-231 cells with anti-IgM in early G1 phase prevents phosphorylation of the retinoblastoma gene product (pRb) and inhibits entry into S phase. Using unsynchronized cells, we previously demonstrated that cyclin A associated and Cdk2-dependent GST-pRb kinase activity were inhibited in WEHI-231 cells treated with anti-IgM. We now show that progression of elutriated early G1 phase WEHI-231 cells from early into late G1 phase is accompanied by an increase in the abundance of cyclin A protein and cyclin A-associated kinase activity. Treatment of early G1 cells with anti-IgM prevented this increase in cyclin A associated kinase activity at late G1, despite minimal changes in the overall level of cyclin A and Cdk2 proteins. Late G1 cells, which already possess high cyclin A-associated kinase activity, were insensitive to anti-IgM treatment and were able to complete the cell cycle. We also found that anti-IgM-treated cells contained increased amounts of the Cdk inhibitor protein p27Kip1. Essentially all of the cyclin A in treated cells was associated with p27, a result which we propose explains the lack of cyclin A/Cdk2 kinase activity. Accumulation of p27 in cyclin A kinase complexes, however, did not decrease the amount of Cdk2 bound to cyclin A. Thus, cross-linking IgM on growth-inhibitable B-cell lymphomas affects cyclin A kinase activity by increasing the levels of p27 in this complex, thus preventing productive pRb phosphorylation and leading to cell cycle arrest and subsequent apoptosis. These results are discussed in terms of the cell cycle restriction points that regulate lymphocyte function, as well as the lineage specific differences in cell cycle control. PMID- 8730100 TI - A mechanism for inhibition of E-cadherin-mediated cell-cell adhesion by the membrane-associated mucin episialin/MUC1. AB - Episialin (MUC1, PEM, EMA, CA15-3 antigen) is a sialylated, membrane-associated glycoprotein with an extended mucin-like ectodomain. This domain mainly consists of 30-90 homologous 20-amino acid repeats that are rich in O-glycosylation sites (serines and threonines). It is likely that this part forms a polyproline beta turn helix. As a result, the ectodomain can protrude more than 200 nm above the cell surface, whereas most cell surface molecules do not exceed a length of 35 nm. Normally, episialin is present at the apical side of glandular epithelial cells. On carcinoma cells, however, it can be strongly overexpressed and it is often present over the entire cell surface. We have previously shown that episialin, if it is interspersed between adhesion molecules, nonspecifically reduces cell-cell and cell-extracellular matrix interactions in vitro and in vivo, presumably by steric hindrance caused by the extreme length and high density of the episialin molecules at the cell surface. To analyze the molecular mechanism for this anti-adhesion effect in more detail, we have now deleted an increasing number of repeats in the episialin cDNA and transfected the resulting mutants into murine L929 cells expressing the homophilic adhesion molecule E cadherin. Here we show that the length of episialin is the dominant factor that determines the inhibition of E-cadherin-mediated cell-cell interactions. For the anti-adhesive effect mediated by the full length episialin, charge repulsion by negatively charged sialylated O-linked glycans is far less important. PMID- 8730102 TI - Construction and characterization of secreted and chimeric transmembrane forms of Drosophila acetylcholinesterase: a large truncation of the C-terminal signal peptide does not eliminate glycoinositol phospholipid anchoring. AB - Despite advances in understanding the cell biology of glycoinositol phospholipid (GPI)-anchored proteins in cultured cells, the in vivo functions of GPI anchors have remained elusive. We have focused on Drosophila acetylcholinesterase (AChE) as a model GPI-anchored protein that can be manipulated in vivo with sophisticated genetic techniques. In Drosophila, AChE is found only as a GPI anchored G2 form encoded by the Ace locus on the third chromosome. To pursue our goal of replacing wild-type GPI-anchored AChE with forms that have alternative anchor structures in transgenic files, we report the construction of two secreted forms of Drosophila AChE (SEC1 and SEC2) and a chimeric form (TM-AChE) anchored by the transmembrane and cytoplasmic domains of herpes simplex virus type 1 glycoprotein C. To confirm that the biochemical properties of these AChEs were unchanged from GPI-AChE except as predicted, we made stably transfected Drosophila Schneider Line 2(S2) cells expressing each of the four forms. TM-AChE, SEC1, and SEC2 had the same catalytic activity and quaternary structure as wild type. TM-AChE was expressed as an amphiphilic membrane-bound protein resistant to an enzyme that cleaves GPI-AChE (phosphatidylinositol-specific phospholipase C), and the same percentage of TM-AChE and GPI-AChE was on the cell surface according to immunofluorescence and pharmacological data. SEC1 and SEC2 were constructed by truncating the C-terminal signal peptide initially present in GPI-AChE: in SEC1 the last 25 residues of this 34-residue peptide were deleted while in SEC2 the last 29 were deleted. Both SEC1 and SEC2 were efficiently secreted and are very stable in culture medium; with one cloned SEC1-expressing line, AChE accumulated to as high as 100 mg/liter. Surprisingly, 5-10% of SEC1 was attached to a GPI anchor, but SEC2 showed no GPI anchoring. Since no differences in catalytic activity were observed among the four AChEs, and since the same percentage of GPI AChE and TM-AChE were on the cell surface, we contend that in vivo experiments in which GPI-AChE is replaced can be interpreted solely on the basis of the altered anchoring domain. PMID- 8730101 TI - Novel syntaxin homologue, Pep12p, required for the sorting of lumenal hydrolases to the lysosome-like vacuole in yeast. AB - pep12/vps6 mutants of Saccharomyces cerevisiae are defective in delivery of soluble vacuolar hydrolases to the vacuole. Morphological analysis by electron microscopy revealed that pep12 cells accumulate 40- to 50-nm vesicles. Furthermore, pep12 cells have enlarged vacuoles characteristic of class D pep/vps mutants. PEP12 encodes a protein of 288 amino acids that has a C-terminal hydrophobic region and shares significant sequence similarity with members of the syntaxin protein family. These proteins appear to participate in the docking and fusion of intracellular transport vesicles. Pep12p is the first member of the syntaxin family to be implicated in transport between the Golgi and the vacuole/lysosome. Pep12p-specific polyclonal antisera detected a 35-kDa protein that fractionated as an integral membrane protein. Subcellular fractionation experiments revealed that Pep12p was associated with membrane fractions of two different densities; the major pool (approximately 90%) of pep12p may associate with the endosome, while a minor pool (approximately 10%) cofractionated with the late Golgi marker Kex2p. These observations suggest that Pep12p may mediate the docking of Golgi-derived transport vesicles at the endosome. PMID- 8730103 TI - Replacement of the glycoinositol phospholipid anchor of Drosophila acetylcholinesterase with a transmembrane domain does not alter sorting in neurons and epithelia but results in behavioral defects. AB - Drosophila has a single glycoinositol phospholipid (GPI)-anchored form of acetylcholinesterase (AChE) encoded by the Ace locus. To assess the role that GPI plays in the physiology, of AChE, we have replaced the wild-type GPI-AChE with a chimeric transmembrane form (TM-AChE) in the nervous system of the fly. Ace null alleles provided a genetic background completely lacking in endogenous GPI-AChE, and Ace minigene P transposon constructs were used to express both GPI- and TM AChE forms in the tissues where AChE is normally expressed. Control experiments with the GPI-AChE minigene demonstrated a threshold between 9 and 12% of normal AChE activity for adult viability. Ace mutant flies were rescued by GPI-AChE minigene lines that expressed 12-40% of normal activity and were essentially unchanged from wild-type flies in behavior. TM-AChE minigene lines were able to rescue Ace null alleles, although with a slightly higher threshold than that for GPI-AChE. Although rescued flies expressing GPI-AChE at a level of 12% of normal activity were viable, flies expressing 13-16% of normal activity from the TM-AChE transgene died shortly after eclosion. Flies expressing TM-AChE at about 30% of normal levels were essentially unchanged from wild-type flies in gross behavior but had a reduced lifespan secondary to subtle coordination defects. These flies also showed reduced locomotor activity and performed poorly in a grooming assay. However, light level and electron microscopic immunocytochemistry showed no differences in the localization of GPI- and TM-AChE. Furthermore, endogenous and ectopic-induced expression of both AChEs in epithelial tissues of the adult and embryo, respectively, showed that they were sorted identically. Most epithelial cells sorted GPI- and TM-AChE to the apical surface, but cuticle-secreting epithelia sorted both proteins basolaterally. Our data suggest that rather than having a primary role in protein sorting, the GPI anchor or AChE plays some other more subtle cellular role in neuronal physiology. PMID- 8730104 TI - Golgi dispersal during microtubule disruption: regeneration of Golgi stacks at peripheral endoplasmic reticulum exit sites. AB - Microtubule disruption has dramatic effects on the normal centrosomal localization of the Golgi complex, with Golgi elements remaining as competent functional units but undergoing a reversible "fragmentation" and dispersal throughout the cytoplasm. In this study we have analyzed this process using digital fluorescence image processing microscopy combined with biochemical and ultrastructural approaches. After microtubule depolymerization, Golgi membrane components were found to redistribute to a distinct number of peripheral sites that were not randomly distributed, but corresponded to sites of protein exit from the ER. Whereas Golgi enzymes redistributed gradually over several hours to these peripheral sites, ERGIC-53 (a protein which constitutively cycles between the ER and Golgi) redistributed rapidly (within 15 minutes) to these sites after first moving through the ER. Prior to this redistribution, Golgi enzyme processing of proteins exported from the ER was inhibited and only returned to normal levels after Golgi enzymes redistributed to peripheral ER exit sites where Golgi stacks were regenerated. Experiments examining the effects of microtubule disruption on the membrane pathways connecting the ER and Golgi suggested their potential role in the dispersal process. Whereas clustering of peripheral pre Golgi elements into the centrosomal region failed to occur after microtubule disruption, Golgi-to-ER membrane recycling was only slightly inhibited. Moreover, conditions that impeded Golgi-to-ER recycling completely blocked Golgi fragmentation. Based on these findings we propose that a slow but constitutive flux of Golgi resident proteins through the same ER/Golgi cycling pathways as ERGIC-53 underlies Golgi Dispersal upon microtubule depolymerization. Both ERGIC 53 and Golgi proteins would accumulate at peripheral ER exit sites due to failure of membranes at these sites to cluster into the centrosomal region. Regeneration of Golgi stacks at these peripheral sites would re-establish secretory flow from the ER into the Golgi complex and result in Golgi dispersal. PMID- 8730105 TI - Heterologous expression of the human cyclin-dependent kinase inhibitor p21Cip1 in the fission yeast, Schizosaccharomyces pombe reveals a role for PCNA in the chk1+ cell cycle checkpoint pathway. AB - Fission yeast cells expressing the human gene encoding the cyclin-dependent kinase inhibitor protein p21Cip1 were severely compromised for cell cycle progress. The degree of cell cycle inhibition was related to the level of p21Cip1 expression. Inhibited cells had a 2C DNA content and were judged by cytology and pulsed field gel electrophoresis to be in the G2 phase of the cell cycle. p21Cip1 accumulated in the nucleus and was associated with p34cdc2 and PCNA. Thus, p21Cip1 interacts with the same targets in fission yeast as in mammalian cells. Elimination of p34cdc2 binding by mutation within the cyclin-dependent kinase binding domain of p21Cip1 exaggerated the cell cycle delay phenotype. By contrast, elimination of PCNA binding by mutation within the PCNA-binding domain completely abolished the cell cycle inhibitory effects. Yeast cells expressing wild-type p21Cip1 and the mutant form that is unable to bind p34cdc2 showed enhanced sensitivity to UV. Cell cycle inhibition by p21Cip1 was largely abolished by deletion of the chk1+ gene that monitors radiation damage and was considerably enhanced in cells deleted for the rad3+ gene that monitors both DNA damage and the completion of DNA synthesis. Overexpression of PCNA also resulted in cell cycle arrest in G2 and this phenotype was also abolished by deletion of chk1+ and enhanced in cells deleted for rad3+. These results formally establish a link between PCNA and the products of the rad3+ and chk1+ checkpoint genes. PMID- 8730106 TI - Evidence that a single monolayer tubulin-GTP cap is both necessary and sufficient to stabilize microtubules. AB - Evidence that 13 or 14 contiguous tubulin-GTP subunits are sufficient to cap and stabilize a microtubule end and that loss of only one of these subunits results in the transition to rapid disassembly(catastrophe) was obtained using the slowly hydrolyzable GTP analogue guanylyl-(a,b)-methylene-diphosphonate (GMPCPP). The minus end of microtubules assembled with GTP was transiently stabilized against dilution-induced disassembly by reaction with tubulin-GMPCPP subunits for a time sufficient to cap the end with an average 40 subunits. The minimum size of a tubulin-GMPCPP cap sufficient to prevent disassembly was estimated from an observed 25- to 2000-s lifetime of the GMPCPP-stabilized microtubules following dilution with buffer and from the time required for loss of a single tubulin GMPCPP subunit from the microtubule end (found to be 15 s). Rather than assuming that the 25- to 2000-s dispersion in cap lifetime results from an unlikely 80 fold range in the number of tubulin-GMPCpP subunits added in the 25-s incubation, it is proposed that this results because the minimum stable cap contains 13 to 14 tubulin-GMPCPP subunits. As a consequence, a microtubule capped with 13-14 tubulin-GMPCPP subunits switches to disassembly after only one dissociation event (in about 15 s), whereas the time required for catastrophe of a microtubule with only six times as many subunits (84 subunits) corresponds to 71 dissociation events (84-13). The minimum size of a tubulin-GMPCPP cap sufficient to prevent disassembly was also estimated with microtubules in which a GMPCPP-cap was formed by allowing chance to result in the accumulation of multiple contiguous tubulin GMPCPP subunits at the end, during the disassembly of microtubules containing both GDP and GMPCPP. Our observation that the disassembly rate was inhibited in proportion to the 13-14th power of the fraction of subunits containing GMPCPP again suggests that a minimum cap contains 13-14 tubulin-GMPCPP subunits. A remeasurement of the rate constant for dissociation of a tubulin-GMPCPP subunit from the plus-end of GMPCPP microtubules, now found to be 0.118 s-1, has allowed a better estimate of the standard free energy for hydrolysis of GMPCPP in a microtubule and release of Pi: this is +0.7 kcal/mol, rather than -0.9 kcal/mol, as previously reported. PMID- 8730107 TI - Hypothalamic-pituitary-adrenal-glucocorticoid axis function in rheumatoid arthritis. PMID- 8730108 TI - The case for a simple method of grading osteoarthritis severity at arthroscopy. PMID- 8730109 TI - Challenges for clinical decision making in the management of osteoarthritis of the knee. PMID- 8730110 TI - Cloning of collagenase 3 from the synovial membrane and its expression in rheumatoid arthritis and osteoarthritis. AB - OBJECTIVE: To analyze synovial membrane of patients with rheumatoid arthritis (RA) for the expression of unknown matrix metalloproteinases (MMP). METHODS: Degenerate oligonucleotides corresponding to highly conserved regions of the MMP gene family and the rapid amplification of cDNA ends (RACE) method have been used to search for new members of this gene family. MMP gene expression has been characterized by Northern blot analysis. RESULTS: We cloned a MMP cDNA from the synovial membrane that is completely identical to the recently published collagenase 3 cDNA derived from a human breast cancer cDNA library (Freije, et al: J Biol Chem 1994;269:16766-73). Collagenase 3 is expressed in parallel with interstitial collagenase and stromelysin 1 in RA and osteoarthritis (OA). Collagenase 3 gene expression was not detected in several normal human tissues. CONCLUSION: The expression of collagenase 3 in the synovial membrane in RA and OA suggests its involvement in articular tissue degradation. PMID- 8730111 TI - Intact adrenocorticotropic hormone secretion but impaired cortisol response in patients with active rheumatoid arthritis. Effect of glucocorticoids. AB - OBJECTIVE: To study the hypothalamic-pituitary-adrenal axis in rheumatoid arthritis (RA) and the influence of glucocorticoid treatment. METHODS: Consecutive untreated patients with RA with moderately high inflammatory activity were studied and compared with healthy subjects of similar age. Subjects were studied both at baseline and after multiple releasing hormone (MRH) stimulations. Patients were reexamined one week after starting prednisolone. RESULTS: The baseline cortisol/adrenocorticotropic hormone (ACTH) ratio was significantly lower in patients with RA. After corticotropin releasing hormone (CRH) stimulation, their serum cortisol response was reduced during the later test phases in spite of intact ACTH response. The baseline and stimulated levels of luteinizing hormone (LH), follicle stimulating hormone (FSH), and thyroid stimulating hormone (TSH) were normal. An impaired prolactin response was seen after MRH stimulation. After one week of prednisolone therapy the absolute response of serum cortisol to CRH was decreased and the stimulated prolactin response was normalized. CONCLUSION: Impaired cortisol secretion in patients with RA in the presence of intact ACTH secretion is consistent with relative adrenal glucocorticoid insufficiency. Adrenal impairment may be secondary to the inflammatory disease process. PMID- 8730112 TI - Low serum creatine kinase activity is associated with muscle weakness in patients with rheumatoid arthritis. AB - OBJECTIVE: In rheumatoid arthritis (RA) serum creatine kinase (CK) is reduced in association with inflammatory response variables. Our objective was to examine whether low CK is associated with muscle weakness and to what extent the hypothesized relationship between CK and muscle weakness can be explained by anthropometric and sociodemographic variables and/or disease variables. METHODS: Cross sectional and longitudinal retrospective analyses of clinical, radiological, and biochemical data of a prospective cohort of consecutive patients with RA. Isometric muscle strength was measured with a validated muscle strength index (MSI); CK was measured with an enzymatic assay (N-acetyl-cysteine, 37 degrees C). RESULTS: 65 patients were enrolled in the study and we obtained complete one year followup data from 47. In cross sectional analysis, CK was a significant, moderate correlate of the MSI (r = 0.43, p < 0.01). CK remained a significant explanatory variable of the MSI in multivariate models that controlled for demographic variables and lean body mass, corticosteroid use, and biochemical, clinical, and radiological disease variables. In longitudinal dichotomous analyses, worsening in CK was weakly but significantly associated with decreased muscle strength, whereas in linear analyses the association did not reach significance. CONCLUSION: In patients with RA, low CK activity is associated with muscle weakness. Demographic, anthropometric, and disease variables related to muscle mass or muscle atrophy explain only part of this association. Our findings support the hypothesis that muscle weakness may be partly caused by a disease related reduction of CK activity independent of muscle atrophy. PMID- 8730113 TI - A cost effectiveness analysis of cyclosporine in rheumatoid arthritis. AB - OBJECTIVE: To perform a cost effectiveness analysis of cyclosporine (CyA) in the treatment of rheumatoid arthritis (RA). METHODS: Five randomized controlled parallel group clinical trials were selected for metaanalysis. A fixed effects model was used to calculate the treatment effects among the studies. An incremental economic analysis was performed from both a societal perspective and the perspective of Ontario Ministry of Health (MOH). A placebo comparison and 2 head to head comparisons were performed. The total treatment cost was calculated for a typical patient based on a modified intent to treat approach modelled over a one year period. RESULTS: CyA produced a 25% or greater improvement in tender joint account in 35% of the patients relative to 17% of patients receiving placebo. There was no significant difference in improvement between CyA and azathioprine (Aza) or D-penicillamine (D-Pen). From the perspective of the Ontario MOH, the annual incremental cost of achieving the same level of improvement between CyA and Aza was found to be $1,473, and between CyA and D Pen, $1,618; the annual incremental cost effectiveness ratio per patient improved of adopting CyA over placebo was $11,547. From a societal perspective, the incremental cost of CyA was $2,886 and $3,731 between Aza and D-Pen, respectively. The annual incremental cost effectiveness ratio against placebo was $20,698. CONCLUSION: Given budgetary constraints on provincial drug plans, guidelines identifying patients in whom the cost effectiveness of CyA may be expected to be most favorable need to be explored. When CyA is the last option available to alleviate RA, whether it is "good value for money" depends upon the importance placed on patient improvement by the patients and/or by society, and on the alternative uses of the same scarce resources. PMID- 8730114 TI - Functional status assessment of physical impairment in a community based population with rheumatoid arthritis: severely incapacitated patients are rare. AB - OBJECTIVES: To determine the severity of impairment due to rheumatoid arthritis (RA) in a community based population. METHODS: The level of impairment in the study group, which represented 88% of the subjects known to have RA in a rural community in Finland, was assessed using the Keitel function test (KFT), a series of 23 range of motion tasks, with score ranging from 96 (worst) to 0. RESULTS: According to the KFT results, 10% of the subjects had severe impairment with a KFT score of 65-96. As to the different Keitel components, the mean hand function index was 22.6 from a possible score of 42, and the mean lower limb function 8.9 out of 44. CONCLUSIONS: Our results showed that severely incapacitated patients with RA are rare in the community. The reasons remain speculative. Multidisciplinary care, including early and active treatment with disease modifying antirheumatic drugs and a high number of orthopedic operations may be an explanatory factor. Total joint replacement surgery of the large joints, performed in 19% of patients, may explain the absence of marked restriction in mobility and the divergence between the limitations in the upper and lower limb functions in this series. PMID- 8730115 TI - Methotrexate in polymyalgia rheumatica: preliminary results of an open, randomized study. AB - OBJECTIVE: Steroids are the only treatment of polymyalgia rheumatica (PMR). We report the effects of methotrexate (MTX) plus prednisone versus prednisone alone in PMR. METHODS: Twenty-four patients with recent onset PMR were studied in a randomized prospective study lasting one year. Patients were given MTX (MTX arm) 10 mg intramuscularly plus prednisone every week, or prednisone alone (Pred arm). After 6 months an attempt was made to stop prednisone, and to use the lowest possible dose over the next 6 months. RESULTS: At the 12th month, all patients were in clinical remission, acute phase reactants were in the normal range in both arms of the study, 6 patients were no longer taking steroids in the MTX arm versus 0/12 in the Pred arm, and the amount of prednisone in the 2 groups was statistically different (1.84 versus 3.2 g; p < 0.0001). In addition, bone mineral density was significantly decreased in the Pred arm, but not in the MTX arm. CONCLUSION: The MTX regimen allowed the use of much less prednisone over one year to obtain full control of PMR with no loss of efficacy. It also allowed sparing of bone in elderly patients at increased risk of osteoporotic fractures. PMID- 8730116 TI - No correlation between prolactin levels and clinical activity in patients with systemic lupus erythematosus. AB - OBJECTIVE: To assess the frequency of hyperprolactinemia and its possible clinical significance in patients with systemic lupus erythematosus (SLE). METHODS: Concentrations of prolactin (PRL) were determined in 63 consecutive patients with SLE by an immunoradiometric assay. Antinuclear antibodies, anti DNA, ribonucleoprotein, Sm, Ro, La, and anticardiolipin antibodies were determined by standard techniques. PRL concentrations were correlated with SLE clinical and serological activity. RESULTS: Elevated serum concentration of PRL (> 20 ng/ml) were found in 10 of 63 (15.9%) patients [21 to 57 ng/ml, mean (SD) 35.6 (13.4)]. No direct correlation between PRL levels and clinical disease activity of SLE was found (Pearson correlation r = -0.186, NS). In addition, hyperprolactinemia was not associated with serological activity. CONCLUSION: Mild hyperprolactinemia is present in a subset of patients with SLE but its clinical significance is still uncertain. PMID- 8730117 TI - Scleroderma fibroblast phenotype is modulated by endothelial cell co-culture. AB - OBJECTIVE: We established a co-culture system to investigate endothelial cell fibroblast interaction in scleroderma (systemic sclerosis, SSc). Such a system allows reciprocal interaction between these cells. The pattern of phenotypic modulation for normal and SSc fibroblasts in co-culture was compared. METHODS: A virally transformed human umbilical vein endothelial cell (HUVEC) derived cell line (1E-7) was cultured on nitrocellulose membrane inserts above dermal fibroblast monolayers. The effect of co-culture on fibroblast number, [3H] thymidine ([3H]-TdR) incorporation, and collagen (type I) production were compared for 10 SSc and 5 control cell lines. Co-culture with the epithelial lines A549 and A431, and nontransformed HUVEC, was also investigated. RESULTS: We observed a statistically significant increase in cell number and a reduction in collagen production for SSc, but not control, fibroblasts co-cultured with endothelial cells. This co-culture also promoted [3H]-TdR incorporation in both SSc and control fibroblasts. While epithelial cell lines did not influence fibroblast cell number, collagen production by SSc fibroblasts was diminished by A549. CONCLUSIONS: Endothelial cell derived soluble factors modulated fibroblast properties in co-culture, and the different response of SSc compared with normal fibroblasts provides further evidence for a link between endothelial and fibroblast dysfunction in this disease. However, similar effects on SSc fibroblast collagen production were also observed for some epithelial cells, suggesting that modulation of fibroblast properties is not restricted to cells of endothelial origin. PMID- 8730118 TI - Isolated pulmonary hypertension in systemic sclerosis with diffuse cutaneous involvement: association with serum anti-U3RNP antibody. AB - OBJECTIVE: To describe a group of patients with systemic sclerosis (SSc) and diffuse cutaneous (dc) involvement with isolated pulmonary hypertension (IPHT) and to compare them to 2 other SSc patient groups, i.e., one with limited cutaneous (lc) involvement with IPHT and another with dcSSc without IPHT. METHODS: The Pittsburgh Scleroderma Databank was screened to identify appropriate patients. SSc specific serum autoantibodies were determined using published methods. Chi-squared tests and Kaplan-Meier survival analysis were performed. RESULTS: During the period 1975-92, 60 of 580 (10%) consecutive new patients with lcSSc and 14 of 677 (2%) with dcSSc have developed IPHT (p < 0.0001). The patients with IPHT did not differ from their parent dc or lc groups by age, race, sex, or cutaneous manifestations. All 14 patients with dcSSc IPHT had minimal or no pulmonary interstitial fibrosis on chest radiograph. The 12 tested had a striking reduction in DLCO (mean 50% of predicted normal), similar to results for patients with lcSSc with IPHT. Of note, 5 of these 14 patients had survived "scleroderma renal crisis" 1.3 to 10.7 (mean 5.5) years before onset of IPHT. Six of 13 (46%) patients with dcSSc with IPHT were positive for serum anti-U3RNP antibody by immunoprecipitation, and one had the typical nucleolar staining pattern of anti-U3RNP, but was not tested for U3RNP by immunoprecipitation. In contrast, only 13 (6%) of the 244 patients with dcSSc without IPHT (p < 0.0001) had anti-U3RNP antibody. Survival from the first reported symptom consistent with pulmonary hypertension (usually dyspnea on exertion) was uniformly poor in both IPHT groups, with the vast majority of patients dying during the first 2 years. CONCLUSION: Patients with SSc with dc involvement can develop severe, fatal isolated pulmonary arterial hypertension. This complication occurs disproportionately more often in patients with serum anti-U3RNP antibody. PMID- 8730119 TI - Twenty-four hour heart period variability in systemic sclerosis. AB - OBJECTIVE: In patients with systemic sclerosis (SSc), an autonomic dysfunction has been implicated in the pathogenesis of Raynaud's phenomenon and gastrointestinal symptoms. To evaluate cardiovascular autonomic function we analyzed the heart rate variability (HRV) on 24 h electrocardiogram recordings. METHODS: In 50 patients diagnosed with SSc and 24 control subjects HRV was analyzed for both frequency and time domain. RESULTS: Frequency domain analysis showed reduction of low frequency and high frequency values in the patients with SSc compared to controls (5.52 +/- 0.74 vs 6.04 +/- 0.78 In(ms2), p = 0.007; and 4.17 +/- 0.97 vs 4.74 +/- 0.97 In(ms2), p = 0.021, respectively). Analysis in time domain showed no significant differences in any variable between groups. There were no significant differences between the diffuse and the limited form of SSc. Number of ventricular ectopic beats, prevalence of frequent ventricular ectopy, and supraventricular tachycardia episodes were increased in the patients with SSc compared to controls. CONCLUSION: Spectral analysis of heart period variability suggests cardiovascular autonomic dysfunction in patients with SSc. Better understanding of this complication could help delineate new strategies for the treatment of SSc. PMID- 8730121 TI - Dietary intake of micronutrient antioxidants in relation to blood levels in patients with systemic sclerosis. AB - OBJECTIVE: To document habitual intakes of micronutrient antioxidants in patients with systemic sclerosis (SSc) in light of studies reporting subnormal levels of ascorbate and selenium in this patient group. METHODS: Dietary intakes of vitamin C, selenium, alpha-tocopherol, beta-carotene, and sulfur amino acid precursors of glutathione were assessed using the 7 day weighed record in 12 patients with SSc and in 12 healthy control subjects. The intakes of the first 4 substances were examined in relation to plasma/serum levels, while intakes of sulfur amino acids were examined in relation to urinary inorganic sulfate. RESULTS: Antioxidant and sulfur amino acid intakes were similar in patients and controls, although the patients had lower levels of selenium (median 74 compared to 87 milligrams in controls; p = 0.014) and of vitamin C in plasma (median 6.0 compared to 11.1 milligrams/l in controls; p = 0.08). Inorganic sulfate concentration in urine was similar in patients and controls. CONCLUSION: Our results suggest that reduced blood levels of the water soluble antioxidants selenium and ascorbic acid in patients with SSc are not due to dietary deficiency. Other explanations must therefore be sought. PMID- 8730120 TI - Soluble serum interleukin 2 receptors in patients with systemic sclerosis. AB - OBJECTIVE: To determine whether soluble interleukin 2 receptor (sIL-2R) levels correlate with natural history of disease in patients with systemic sclerosis (SSc). METHODS: The following groups were studied: Group I included 81 consecutive new patients with SSc. Group II consisted of 21 patients with diffuse cutaneous (dc) SSc whose peripheral blood and affected skin had been analyzed for T lymphocyte subsets. Group III contained 38 patients with dcSSc with serial sIL 2R determinations during the course of disease. sIL-2R was performed using a commercial double monoclonal antibody ELISA technique. RESULTS: The 81 Group I patients with SSc had a mean sIL-2R level of 821 units compared with 35 controls, who had a mean of 389 units (p < 0.001). sIL-2R level significantly correlated with the extent of skin thickening (p < 0.005). In Group II patients, sIL-2R was found to correlate with the CD4 to CD8 ratio. Blindly assessed clinical evidence of disease activity from the serial samples of 38 Group III patients was consistent with sIL-2R levels in 83% of the samples. There was high correlation of change in sIL-2R with change in skin score over time in Group III subjects (r = 0.71). CONCLUSIONS: These results suggest a role for T cell activation in the pathogenesis of SSc. sIL-2R levels may be useful adjunct to clinical evaluation in assessing disease activity and predicting future events in patients with SSc. PMID- 8730122 TI - A multicenter trial of recombinant human interferon gamma in patients with systemic sclerosis: effects on cutaneous fibrosis and interleukin 2 receptor levels. AB - OBJECTIVE: To evaluate the acute toxicity, potential efficacy, and effects on the soluble interleukin 2 receptor (sIL-2R) of recombinant human interferon gamma (rIFN-gamma) in patients with systemic sclerosis (SSc). METHODS: A multicentered, pilot clinical trial of rIFN-gamma was performed on 20 patients (15 women, 5 men, mean age 45 years) with active cutaneous SSc (mean disease duration 36 months) to evaluate it potential as a novel therapy for this untreatable disorder. After one week of rIFN-gamma 0.01 mg/m2/day, subjects self-administered rIFN-gamma 0.1 mg/m2/day intramuscularly for a total of 18 weeks. The major outcome variable was a modified skin score (0 = normal skin, 3 = hidebound skin) measured and summed from 15 anatomic areas of the body. sIL-2R levels were measured by ELISA at entry and exit from the study. RESULTS: The clinical results were modest at best. Nine of 20 patients achieved at least a 20% reduction in skin score, with one patient showing almost total remission of all skin abnormalities. The mean skin score at entry for all subjects was 22.8 +/- 8.9 and over the course of the trial improved marginally compared to baseline (mean change -4.72 +/- 6.62; p = 0.008). However, 8 subjects did not change appreciably while in the trial. Antibodies to Scl-70 were observed in only 5 patients (all with diffuse scleroderma) and were not associated with either response to or complications from therapy. The adverse reactions were frequent and occasionally severe. Ten subjects were withdrawn because of exacerbation of Raynaud's symptoms (n = 5), constitutional symptoms (n = 2), development of renal crises (n = 2), and mild pancytopenia (n = 1). Minor laboratory abnormalities were common and included elevation of cholesterol, triglycerides, hepatic transaminases, and reduction in white blood cell count. Compared to controls, mean sIL-2R was markedly elevated at entry (1309 +/- 495 U/ml; p = 0.0001) and did not change appreciably at exit. Spearman correlation analysis showed a trend but no statistically significant association of skin score with sIL-2R (R = 0.408; p = 0.074). However, sIL-2R was significantly correlated with erythrocyte sedimentation rate (R = 0.542; p = 0.0165). A subset analysis revealed that skin score (p = 0.0001) and sIL-2R (p = 0.00170) were significantly higher at baseline for patients with diffuse scleroderma compared to patients with limited disease. CONCLUSION: rIFN-gamma may be beneficial for some patients with SSc, but the benefit appears marginal for most individuals and the side effects frequent. Although sIL-2R was elevated in many of the patients with SSc, it did not appear to be correlated with activity of cutaneous disease or response to therapy. PMID- 8730124 TI - Inorganic pyrophosphate generation from adenosine triphosphate by cell-free human synovial fluid. AB - OBJECTIVE: To quantify inorganic pyrophosphate (PPi) production from extracellular adenosine triphosphate (ATP) by human synovial fluids (SF). METHODS: Serial measurements of ATP hydrolysis rate (t1/2) were performed by the luciferase method from a starting concentration of 500 nM in 21 pathologic and one normal cell-free SF samples incubated under physiologic conditions. ATP was then pumped into a sample of each fluid, using the rate constant derived from the t1/2 of that fluid, to provide steady state levels simulating those reported in SF. Trace [32P] gamma ATP was added at the start of the infusion; conversion to [32P] Pi and to [32P] PPi was determined by precipitation of Pi as reduced phosphomolybdate before and after treatment with yeast inorganic pyrophosphatase. Finally, the pumping experiment was repeated and PPi production was calculated from direct measurement of PPi at time zero and at 60 min. PPi hydrolysis was measured in each fluid by [32P] Pi precipitation from [32P] PPi tracer added at time zero. RESULTS: ATP was hydrolyzed by all SF. The mean t1/2 (seconds) in 8 osteoarthritis (OA) samples was 72 s, in 5 calcium pyrophosphate dihydrate (CPPD) 30 s (p < 0.02), in 3 rheumatoid arthritis (RA) 1160 s, in normal 86 s, in 3 olecranon bursal (OB) 54 s, and in 2 total knee replacement fluid samples 17 and 121 s. The major product of ATP hydrolysis was PPi in all but 2 fluids (1 RA, 1 OB), even at lower than steady state levels. At simulated in vivo steady state ATP levels, mean conversion of APT to PPi was stoichiometric in OA and CPPD fluids. PPi hydrolysis was < 4% in all noninflammatory fluids. CONCLUSION: PPi is the major product of extracellular ATP catabolism in most SF. Hydrolysis rates were significantly faster in SF containing CPPD crystals. Mean PPi production by these fluids at simulated in vivo steady state levels was 6-fold that of OA SF (p < 0.01). Hydrolysis of extracellular ATP by ectonucleotide pyrophosphohydrolases can account for all PPi produced by joint issues previously estimated from [32P] PPi pool and turnover studies in human knee joints. PMID- 8730123 TI - Computed tomography guided corticosteroid injection of the sacroiliac joint in patients with spondyloarthropathy with sacroiliitis: clinical outcome and followup by dynamic magnetic resonance imaging. AB - OBJECTIVE: To evaluate computed tomography (CT) guided corticosteroid injections of inflamed sacroiliac (SI) joints in patients with spondyloarthropathies (SpA), and to evaluate dynamic magnetic resonance imaging (DMRI) of the SI joints in serial examinations of these patients, who had different degrees of inflammatory back pain. METHODS: We examined and treated 30 patients with ankylosing spondylitis (n = 9) or undifferentiated SpA (n = 11) (14 women and 16 men, mean age 36.5 +/- 13.4 years, mean disease duration 5.4 +/- 4.0 years) who had severe inflammatory back pain for more than 3 months. All patients had DMRI of the SI joints before and 4-6 months after a CT guided injection of 40 mg triamcinolone acetonide into SI joints (n = 54; 24 patients received injections in both joints). Enhancement of the contrast agent gadolinium-DTPA was quantified by calculating the enhancement of the contrast agent gadolinium-DTPA was quantified by calculating the enhancement gradient Fenh. A subjective index with a visual analog scale (0 = no pain, 10 = very severe pain) was used for assessment of back pain. Followup visits were done every 3 months for a maximum of 18 months. RESULTS: There was significant improvement of inflammatory back pain and sacroiliitis at 5.2 +/- 1.3 months after therapy in 25/30 patients (83.3%). The differences between the Fenh values before (98.2 +/ 56.1) and after (44.3 +/- 31.2) therapy and of the subjective pain index (8.5 +/- 1.5 and 3.0 +/- 2.3, respectively) were statistically significant. Subjective improvement lasted a mean of 8.9 +/- 5.3 months. CONCLUSION: CT guided corticosteroid injection of inflamed SI joints is a useful option in therapy for sacroiliitis in patients with SpA. Different degrees of inflammation in the SI joints can be quantitatively assessed by DMRI. PMID- 8730125 TI - A metabolic syndrome in diffuse idiopathic skeletal hyperostosis. A controlled study. AB - OBJECTIVE: I addition to diabetes mellitus (DM), high incidence of common metabolic conditions such as dyslipidemia (DL) and hyperuricemia (HU) has been found in patients with diffuse idiopathic skeletal hyperostosis (DISH). Our objective was to confirm such observations comparing data from patients with DISH and appropriate controls. METHODS: One hundred randomly identified inpatients with DISH were compared with 100 DISH-free inpatients with various rheumatic disorders matched for age, sex, body mass index, and excess body weight. The prevalence of metabolic disturbances was compared in the 2 groups. Data was analyzed statistically by chi-squared test and 4-fold table analysis. RESULTS: In our DISH population, metabolic abnormalities were found in 70%, versus 45% in the control population (p < 0.0001). High prevalence of DL and/or HU associated with DM was observed among patients with DISH (p < 0.0001). Patients with uncomplicated diabetes (or DL or HU) were almost equally distributed between the 2 groups, with no significant differences (p > 0.1). Combined metabolic features of DM+DL, DM+HU, or DM+DL+HU were shown to be the main risk factor for DISH (K = 14). They may constitute a metabolic syndrome with high diagnostic specificity (u = 1.0) and nosologic sensitivity (g = 1.0), despite their lowest prevalence (0.07). Dyslipidemia occurred with the highest prevalence (p = 0.36), the highest positive predictive value (v = 0.44) and, together with HU and diabetes, the highest nosologic specificity (f = 0.55 in both cases). CONCLUSION: We found metabolic disturbances confined to the group with DISH. Followup studies of the number of bridging ossifications in identically aged patients or bridging ossifications/year/patient could be a useful tool to determine their effect on the extent and progression rates of DISH. PMID- 8730126 TI - Regulation of alkaline phosphatase: implications for calcium pyrophosphate dihydrate crystal dissolution and other alkaline phosphatase functions. AB - OBJECTIVE: Alkaline phosphatase (ALP), an enzyme with pyrophosphatase (PPiase) activity can dissolve calcium pyrophosphate dihydrate (CPPD) crystals. We studied the effects of enzyme inhibitors such as bisphosphonates, orthovanadate, calcium, cadmium, and ascorbic acid on PPiase activity of ALP as well as on phosphate ester hydrolysis (Pase) activity and compared these effects to those on CPPD crystal dissolution. METHOD: An in vitro model system for crystal enzyme interaction was used to assess CPPD crystal dissolution. RESULTS: Bisphosphonates inhibited ALP Pase activity more than ALP PPiase activity at the same concentrations. Calcium inhibited ALP PPiase activity, but not ALP Pase activity. Orthovanadate and cadmium inhibited ALP PPiase activity more than ALP Pase at the same concentrations. The inhibition rates of ALP PPiase at the same concentrations were orthovanadate > cadmium > calcium. Although ALP Pase activity was not inhibited, at high concentrations, ascorbic acid slightly inhibited ALP PPiase activity. Bisphosphonates at high concentrations inhibited ALP CPPD crystal dissolution. The strong inhibitory effects of bisphosphonates on ALP CPPD crystal dissolution compared to those on ALP PPiase activity suggest that bisphosphonates inhibit crystal dissolution by their affinity for the CPPD crystal surface. Calcium, orthovanadate, and cadmium inhibited ALP CPPD dissolution. The inhibition rates of ALP CPPD dissolution at the same concentrations were cadmium > calcium > orthovanadate. Ascorbic acid at high concentrations enhanced ALP CPPD dissolution. CONCLUSION: These effects of different inhibitors on ALP PPiase and CPPD dissolution suggest that ALP CPPD crystal dissolution depends on binding of ALP CPPD crystals as well as the PPiase activity of the bound ALP. Because of its ubiquitous and broad phosphatase activity including PPiase activity, ALP may have a critical role in cell energy metabolism. PMID- 8730127 TI - Gouty arthritis in nodal osteoarthritis. AB - OBJECTIVE: To analyze the clinical features and identify factors associated with the development of gouty arthritis in nodal osteoarthritis (OA). METHODS: Thirty two consecutive patients (21 women and 11 men, mean age 75.8 years) with both nodal OA and crystal proven acute gout and/or tophi of distal/proximal interphalangeal (DIP/PIP) joints were studied between 1986 and 1994. RESULTS: Tophi of DIP and/or PIP joints were present in 29 (90%) patients; alone in 9 and together with acute DIP or PIP gouty arthritis in 20. Three patients had acute DIP or PIP gouty episodes but no digital tophi. Mean pretreatment serum urate was 614.9 +/- 163.2 (range 422-1088 mumol/l). Risk factors for gout included diuretic use (81%), renal failure (59%), hypertension (66%), alcoholism (22%), prophylactic low dose ASA (20%), and a positive family history (16%) of patients. CONCLUSION: The coexistence of gouty arthritis in nodal OA is important to recognize and treat, particularly in elderly women with renal failure, hypertension, or cardiac failure who are receiving longterm diuretic therapy. PMID- 8730128 TI - Finger pad tophi in gout. AB - OBJECTIVE: To determine frequency and prevalence of finger pad tophaceous deposits in patients with chronic tophaceous gout. METHODS: We examined 36 consecutive patients with chronic tophaceous gout for intracutaneous tophaceous urate deposits in the fingers pads. Subjects were patients of outpatient clinics at 2 large university hospitals, Emory and the University of Pennsylvania Affiliated Hospital Systems. RESULTS: Finger pad tophaceous deposits were found in 30.5%, which was surprising, given how few cases are reported. CONCLUSION: Finger pad tophi may be more common than generally believed. PMID- 8730129 TI - Severity of radiographic findings in hip osteoarthritis associated with total hip arthroplasty. AB - OBJECTIVE: The decision to perform total hip arthroplasty (THA) in patients with osteoarthritis (OA) of the hip is based largely on patients' reports of pain and disability and not on radiographic findings of OA. We determine the severity of radiographic OA and its association with disability in patients undergoing THA. METHODS: Individual radiographic features (osteophytes, joint space narrowing, sclerosis, cysts, deformity) and global severity of hip OA were assessed in 95 consecutive elderly patients with hip OA undergoing THA who were enrolled in a Patient Outcome Research Team (PORT) project. RESULTS: Eighty-seven patients (91.5%) had either severe or moderate OA in the hip to be replaced; 17% of these had a previous contralateral THA. Only 8 patients (8.4%) had mild or no signs of OA in the hip to be replaced and 4 (50%) of these patients had their opposite hip replaced previously. CONCLUSION: These data indicate that radiographic features of moderate to severe hip OA are associated with clinical findings and the necessity to perform THA in the majority of patients. Patients who have had a prior hip replacement, however, may be more likely to have a contralateral replacement done earlier (p = 0.03), before radiographic signs are evident. PMID- 8730130 TI - Arthroscopic evaluation of chondropathy in osteoarthritis of the knee. AB - OBJECTIVE: To evaluate arthroscopic findings as a potential measurement of severity and outcome of chondropathy. METHODS: 110 patients with knee osteoarthritis (OA) were included in a cross sectional study and 41 of them in a one year longitudinal study. The evaluation of OA performed once in the cross sectional study and twice (at entry and after one year) in the longitudinal study, included clinical, radiological and arthroscopic variables evaluating disease activity and severity. Arthroscopy was performed. under local anesthesia in an outpatient procedure using a small arthroscope. Chondropathy was evaluated by the overall assessment of the investigator by visual analog scale and the Societe Francaise d'Arthroscopie (SFA) scoring and grading systems, which represent a composite index taking into account depth, size, and localization of the articular cartilage lesions. RESULTS: The intrinsic validity of the arthroscopic variables was suggested by highly significant correlation (R2 = 80 85%) between the overall assessment of the investor and the SFA systems. There was also highly significant correlation (p < 0.01) between the arthroscopic and radiological variables. Intraobserver reliability of the arthroscopic quantification of chondropathy was better than interobserver reliability. In the cross sectional study, severity of chondropathy correlated with both age and body mass index. In the longitudinal study there was statistically significant worsening in the severity of chondropathy and statistically significant correlation between the changes in the severity of chondropathy and changes in functional impairment. CONCLUSION: We conclude that arthroscopy might be considered a relevant measurement of OA outcome for research purposes. PMID- 8730131 TI - The effect of arthritis on the quality of life in hereditary hemochromatosis. AB - OBJECTIVE: To assess clinical factors affecting the quality of life in patients with hereditary hemochromatosis. METHODS: Fifty consecutive patients with hereditary hemochromatosis completed a self-administered survey (Medical Outcome Survey, MOS-SF36). The effects of cirrhosis, diabetes, and arthritis on physical, social, and general health variables were assessed in 8 subscales including physical, social, emotional, and general health. RESULTS: Multiple linear regression models demonstrated that arthritic patients had impairments in more subcategories (physical functioning, pain and general health perception) than cirrhotic (vitality) and diabetic patients. A multivariate analysis of variance model suggested that arthritis is the strongest single factor affecting quality of life (p = 0.0516). CONCLUSION: Although cirrhosis is the major factor affecting survival, arthritis is a prominent clinical factor affecting quality of life in hemochromatosis. PMID- 8730132 TI - Lack of association between thyroid status and chondrocalcinosis or osteoarthritis: the Framingham Osteoarthritis Study. AB - OBJECTIVE: To assess whether current thyroid status is associated with chondrocalcinosis or osteoarthritis (OA), we examined the cross sectional association of serum thyrotropin (thyroid stimulating hormone, TSH) with chondrocalcinosis and with knee OA in members of the Framingham OA study. METHODS: Knee radiographs were taken at the 18th biennial examination (1983-85) and measurement of serum TSH at either the 15th (1977-79) or the 18th biennial examination. Chondrocalcinosis was dichotomized as absent or present and knee OA was based on a Kellgren and Lawrence score of grade 2 or greater in either knee. Thyroid status was determined by serum TSH concentration classed into clinically relevant categories: < or = 0.1 mU/l(low); > 0.1 < 0.4 mU/l (slightly low); 0.4 to 5.0 mU/l (normal); > 5 < or = 10 mU/l (slightly high); and > 10 mU/l (high). RESULTS: Data were collected on 577 men and 798 women. We found no association between elevated serum TSH concentration and chondrocalcinosis. Our results, though not statistically significant, suggest an inverse relation, with an odds ratio (OR) of 0.41 (95% CI 0.10, 1.73) for those subjects. in the highest TSH group and 1.79 (95% CI 0.39, 8.24) for those in the lowest TSH group, compared to subjects in the normal range. We found no association between serum TSH concentration and radiographic knee OA, with an OR of 0.85 (95% CI 0.47, 1.51) for those in the highest serum TSH group and 1.51 (95 CI 0.54, 4.22) for those in the lowest TSH group, compared to the normal group. Exclusion of subjects taking thyroid hormone confirmed these null results. CONCLUSION: There was no evidence, in a large unselected population of older persons, of a significant association between current thyroid status and either chondrocalcinosis or OA. PMID- 8730133 TI - The effect of noninsulin dependent diabetes mellitus on the prevalence of clinical osteoarthritis. A population based study. AB - OBJECTIVE: To explore the relation between noninsulin dependent diabetes mellitus (NIDDM) and osteoarthritis (OA) in a population. METHODS: The study population included 632 men and 882 women aged 52-95 years from the Rancho Bernardo community. In 1984-87, participants answered questions about history of diabetes and had a standard oral glucose tolerance (OGTT). In 1988-92, subjects completed a questionnaire about history of arthritis, type of arthritis diagnosed, and presence of joint pain. Nurses examined subjects for presence of Heberden's nodes. Subjects with no history of arthritis were compared to those with a history of OA and other types of arthritis with regard to age, body size, and plasma glucose levels. In addition, subjects were classified by diabetes status to determine differences in the prevalence of arthritis and related characteristics. RESULTS: Neither impaired glucose tolerance nor NIDDM was associated with history of OA, regardless of how inclusive the definition of OA, before or after adjustment for age and maximum lifetime obesity. In age and obesity adjusted analyses, men with a history of OA had lower fasting plasma glucose levels than men with no arthritis (100.2 vs. 103.6 mg/dl, p < 0.05), and men with NIDDM had less hand and hip pain than normoglycemic men (p < 0.05). Heberden's nodes were unrelated to glucose tolerance status. CONCLUSION: This population based study found no positive association between clinical OA and NIDDM defined by OGTT. These results are compatible with community based data examining radiographic OA and history of diabetes. PMID- 8730134 TI - Aceclofenac, a new nonsteroidal antiinflammatory drug, decreases the expression and function of some adhesion molecules on human neutrophils. AB - OBJECTIVE: To study the effect of aceclofenac, a new nonsteroidal antiinflammatory drug (NSAID), on the expression and function of adhesion molecules in human neutrophils. METHODS: We used flow cytometry analysis to determine peripheral blood neutrophil expression of L-selectin, CD11a, CD11b, CD31, CD43, CD44, and intercellular adhesion molecule 2 (ICAM-3) surface adhesion molecules after treatment with aceclofenac, diclofenac, or dexamethasone. Granular enzyme activity was quantitated in extracellular medium of neutrophils treated with different NSAID: In vitro adhesion assays were developed to examine the effects of aceclofenac on both neutrophil adhesion to tumor necrosis factor alpha stimulated human umbilical vein endothelial cells under nonstatic conditions, and homotypic neutrophil aggregation induced by anti-ICAM-3 and anti CD18 monoclonal antibodies (Mab). RESULTS: Aceclofenac induced a dramatic decrease of L-selectin expression, whereas a moderate and slight decrement of CD43 and ICAM-3 expression was also observed. In contrast, the expression of other adhesion molecules by neutrophils was unaffected (CD11a, CD31, CD44) or slightly increased (CD11b). Cell adhesion assays, performed under nonstatic conditions, revealed that aceclofenac significantly diminished the L-selectin dependent neutrophil adhesion to endothelial cells. Neutrophil aggregation induced with anti-CD43 Mab was also significantly inhibited by aceclofenac. CONCLUSION: Aceclofenac had a faster and more potent effect than the other NSAID studied, mainly on the expression of cell adhesion molecules. This new NSAID efficiently interferes with neutrophil adhesion to endothelium and this effect may represent an additional relevant mechanism in its antiinflammatory activity. PMID- 8730135 TI - Variation in rheumatologists' and family physicians' perceptions of the indications for and outcomes of knee replacement surgery. AB - OBJECTIVE: To assess agreement among rheumatologists and family physicians (FP) about the indications for knee replacement (KR) referral, use of nonsurgical management options, and perceived outcomes of KR, and to determine the relationship between these opinions and the number of patients seen with severe osteoarthritis (OA) of the knee. METHODS: 98 adult rheumatologists and a random sample of 250 FP in Ontario, Canada were surveyed. Of the practising and traceable rheumatologists and FP, 70.0 and 5.16% responded, respectively. RESULTS: FP disagreed on how 28 of 32 patient factors affected their KR referral decision, while rheumatologists disagreed on 26 of these 32 factors (p = 0.03). Rheumatologists and FP consistently disagreed on the use of 8 of 10 treatments for knee OA (p = 0.37). While rheumatologists and FP reported similar KR outcomes, FP were less in agreement (p = 0.03). Clinical disagreement for the indications for KR (p < 0.0001) and KR outcomes (p < 0.0001) were greater among FP than among orthopedic surgeons who were surveyed in a prior study. Clinical disagreement about the indications for KR was greater among rheumatologists than among surgeons (p = 0.04), but there was no difference in perceived KR incomes (p = 0.18). CONCLUSION: Referring physicians disagreed on the indications for KR referral an on the treatments for knee arthritis, but were in general agreement regarding KR outcomes. Clinical disagreement was greater among FP than among rheumatologists, who in turn reported more disagreement than orthopedic surgeons. Explanations for these difference in perceptions should be the focus of research, but guidelines specifically tailored for each physician specialty may be required to reduce clinical uncertainty. PMID- 8730136 TI - Insulin-like growth factor-I (somatomedin C) levels in chronic fatigue syndrome and fibromyalgia. AB - OBJECTIVE: Fibromyalgia (FM) and chronic fatigue syndrome (CFS) are similar conditions characterized by substantial fatigue, diffuse myalgias, sleep disturbances and a variety of other symptoms. Many patients with CFS meet strict criteria for FM. Recently, low insulin-like growth factor-I (IGF-I) levels have been demonstrated in patients with FM, suggesting that disruption of the growth hormone-IGF-I axis might explain the link between the muscle pain and poor sleep. Our goal was to determine whether IGF-I levels are decreased in CFS, and whether such findings are restricted to patients with concurrent FM. METHODS: Radioimmunoassays were used to determine serum concentrations of IGF-I and its binding protein, (IGFBP-3). Subjects were 3 patients seen in a referral clinic for chronic fatigue: 15 patients with CFS, 15 who met criteria for both CFS and FM (CFS-FM), 27 with FM alone; and 15 healthy control (HC) subjects. RESULTS: Patients and control subjects had similar demographic and clinical characteristics. No significant differences were observed among any of the 3 patient groups and control subjects in the mean concentration of either IGF-I or IGFBP-3. Likewise, the proportion of subjects with values above or below the laboratory's reference range did not differ for IGF-I or IGFBP-3. CONCLUSIONS: These findings suggest the disruption of the growth hormone-IGF-I axis previously demonstrated in FM patients is not evident in a referral population of patients with CFS, CFS-FM, or FM. PMID- 8730137 TI - Biologic agents in autoimmune diseases. PMID- 8730138 TI - Polymorphism in the LMP2 gene influences disease susceptibility and severity in HLA-B27 associated juvenile rheumatoid arthritis. AB - OBJECTIVE: To determine the potential contribution of the MHC class II region proteasome subunit gene, LMP2, to disease susceptibility, severity, and phenotype in patients with juvenile rheumatoid arthritis (JRA). METHODS: A CfoI restriction site polymorphism in the coding region of the LMP2 gene was evaluated in 279 patients with JRA and 107 healthy controls of similar ethnicity. Patients were divided into 5 groups on the basis of clinical presentation; 46% had early onset pauciarticular disease, 10% early onset polyarticular, 10% late onset pauciarticular, 20% late onset polyarticular, and 11% systemic onset arthritis. The influence of this LMP2 polymorphism on susceptibility to disease, clinical subtype of disease at onset (age and number of joints involved), progression and severity of joint disease (pauci to polyarticular course and radiographic changes), and occurrence of inflammatory eye disease was evaluated. RESULTS: Comparison of genotypes revealed a significantly increased prevalence of homozygosity for the LMP2 B allele (LMP2 BB genotype) in patients who were older (> or = 6 years) at onset of disease (65%, p < 0.05), particularly in those with pauciarticular (71%) involvement at presentation (p < 0.05), compared to controls (51%). The BB genotype was also more prevalent in patients with a polyarticular course, either from onset (63%) or those who progressed from pauciarticular disease (69%), compared with controls, (p = 0.05 and < 0.05, respectively). Stratification for HLA-B27 and DR4, the HLA alleles most frequently associated with late onset pauciarticular and late onset polyarticular JRA, respectively, revealed a persistent effect of LMP2 BB homozygosity on disease susceptibility and phenotype that remained statistically significant in HLA-B27 positive children, and was not due to linkage disequilibrium. CONCLUSION: We show that homozygosity of the B allele of the proteasome subunit LMP2 increases susceptibility to certain subgroups of JRA, and influences the phenotype of disease, predisposing to more progressive and severe articular disease. PMID- 8730139 TI - Pneumocystis carinii pneumonia in childhood systemic lupus erythematosus. AB - OBJECTIVE: Although Pneumocystis carinii pneumonia (PCP) is known to occur in adults with systemic lupus erythematosus (SLE), this infection has rarely been described in childhood SLE. We describe 3 children with SLE who developed PCP and describe risk factors for this complication. METHODS: A retrospective case review. RESULTS: All 3 children had severe active SLE with organ involvement requiring immunosuppressive therapy, but the clinical presentations of PCP differed in each patient. They shared some of the known risk factors for opportunistic infection in adults with SLE, including lymphopenia, but severe lymphopenia (< 0.35 x 10(9)/1) was not seen. CONCLUSION: PCP is an uncommon but serious complication of childhood SLE, and should be considered in the presence of respiratory symptoms, however subtle. The role of oral chemoprophylaxis is discussed. PMID- 8730140 TI - Abdominal aortic aneurysm associated with childhood sarcoidosis. AB - Childhood sarcoidosis is a rare disease with multisystemic organ involvement. A wide spectrum of vasculitides, including large vessel involvement, have been reported in childhood sarcoidosis. We describe a 7-year-old boy with childhood sarcoidosis who presented with prolonged fever, iritis, and abdominal aortic aneurysm. He underwent successful surgical repair of the abdominal aortic aneurysm and followup for 12 months was uneventful. This is first report of abdominal aortic aneurysm complicating childhood sarcoidosis. PMID- 8730141 TI - Rheumatoid papules in a patient with acquired immune deficiency syndrome and symmetric polyarthritis. AB - We describe a patient with acquired immunodeficiency syndrome (AIDS) and an unusual rheumatoid factor-positive symmetric inflammatory polyarthritis who met all 7 American College of Rheumatology criteria for RA, but who also had many features suggestive of seronegative inflammatory arthritis. Although cutaneous vasculitis has also been described in human immunodeficiency virus infection, this is the first report of cutaneous extravascular necrobiotic granuloma (rheumatoid papule) in AIDS. This case highlights the immunopathogenesis of inflammatory arthritis in AIDS and illustrates the difficulties in making a classic rheumatic diagnosis in the setting of the immune dysregulation caused by AIDS. PMID- 8730142 TI - Resolution of the neutropenia of Felty's syndrome by longterm administration of recombinant granulocyte colony stimulating factor. AB - Felty's syndrome is characterized by neutropenia, splenomegaly, and recurrent infection in patients with rheumatoid arthritis. We used recombinant granulocyte colony stimulating factor (rGCSF) in a patient with Felty's syndrome and recurrent sepsis. rGCSF induced a statistically significant increase in the patient's absolute neutrophil and total white blood cell counts. During 14 months of followup taking rGCSF, disseminated varicella zoster was the only infectious complication. Except mild thrombocytopenia and a transient flare of arthritis, no serious adverse effects occurred. rGCSF may be a safe and effective therapy for Felty's syndrome in selected patients. PMID- 8730143 TI - Dapsone syndrome in cutaneous lupus erythematosus. AB - Dapsone is increasingly used in the treatment of rheumatic diseases. It is indicated in many inflammatory skin conditions characterized by polymorphonuclear cell infiltration. Minor reactions to the drug are very common, but severe idiosyncratic reactions are rare considering the large number of patients taking this drug worldwide. We describe 2 patients with cutaneous lupus erythematosus who developed severe dapsone reaction after low dose therapy, with a fatal outcome in one. Physicians should be aware of the potentially lethal side effects of dapsone. PMID- 8730144 TI - Thrombocytopenia corrected by warfarin in antiphospholipid syndrome. AB - We describe a case of antiphospholipid associated thrombocytopenia in which warfarin, but not heparin, normalized the platelet count. Systemic lupus erythematosus was present in association with chorea, mitral valve stenosis, and renal thrombotic microangiopathy. No evidence of platelet consumption was detected. Low dose aspirin, pulse methylprednisolone, intravenous gammaglobulin, and cyclophosphamide also partially elevated the platelet count. Platelet activation, measured by P selectin assay, was not present during warfarin or aspirin therapy, suggesting that platelet activation may have been decreased by these treatments. We discuss possible mechanisms for correction of thrombocytopenia in antiphospholipid syndrome by warfarin. PMID- 8730145 TI - Primary antiphospholipid syndrome presenting as chronic thromboembolic pulmonary hypertension. Treatment with thromboendarterectomy. AB - Pulmonary hypertension may occur in the antiphospholipid syndrome as a result of recurrent pulmonary embolism or microthrombosis of pulmonary vessels. We describe 3 cases of primary antiphospholipid syndrome (APS) and cor pulmonale that fulfilled the criteria for chronic major vessel thromboembolic pulmonary hypertension. Pulmonary thromboendarterectomy was performed in all 3 patients and it was successful in 2. One patient died in the immediate postoperative period from hemorrhagic pulmonary edema. Chronic thromboembolic pulmonary hypertension should be added to the list of pulmonary vascular complications of primary APS. Despite its risk, pulmonary thromboendarterectomy represents a treatment option for this otherwise lethal condition. PMID- 8730146 TI - Scleroderma and watermelon stomach. AB - Two patients with scleroderma (systemic sclerosis, SSc) and chronic blood loss secondary to gastric vascular ectasia--"watermelon stomach"--are presented. These cases exemplify the condition of gastric vascular ectasia and highlight the increasingly recognized association with autoimmune antibodies and connective tissue disease such as SSc. In both cases the onset of gastric blood loss coincided with the clinical manifestations of sclerodactyly by months, suggesting some temporal relationship. Although a number of different treatment modalities are available, and discussed in this report, both cases required gastrectomy. PMID- 8730147 TI - Retroperitoneal fibrosis with antineutrophil cytoplasmic antibodies. AB - A middle aged man showed retroperitoneal fibrosis with signs of vasculitis verified by biopsy of the retroperitoneal mass. Antineutrophil cytoplasmic antibodies (ANCA) with cytoplasmic staining pattern (cANCA) were strongly positive, although there were no clinical or histological signs of Wegener's granulomatosis. No cases of cANCA associated retroperitoneal fibrosis have been described except in connection with Wegener's granulomatosis. ANCA have been found to associate with any inflammatory disorders. This observation may broaden the clinical spectrum of cANCA positive vasculitides. PMID- 8730148 TI - Polymyositis, arthritis, and proteinuria in a patient with adult celiac disease. AB - A 37-year-old woman developed polymyositis and arthritis concomitantly with proteinuria and watery diarrhea. Repeated duodenal biopsies and serological evaluation established the diagnosis of adult celiac disease. Treatment with gluten-free diet resolved all clinical and laboratory abnormalities. We believe that this is the first report of adult celiac disease presenting as a multisystem disease involving kidneys, joint, and muscles. PMID- 8730149 TI - Fulminant hepatic failure in adult onset Still's disease. AB - Adult onset Still's disease (AOSD) is a well characterized rheumatoid disorder with frequent liver involvement. This is usually asymptomatic but severe hepatic failure has occasionally been reported. We describe a 44-year-old woman who developed acute liver failure 2 months after presenting symptoms of AOSD. Full etiologic evaluation was done and all other causes of liver failure were excluded. She underwent emergency orthotopic liver transplantation but developed disseminated intravascular coagulation with acute renal failure, seizures, and coma, and died after 48 hours. PMID- 8730150 TI - Clinically important change in rheumatoid arthritis. PMID- 8730151 TI - Circulating endothelin-1 levels in systemic sclerosis subsets--a marker of fibrosis or vascular dysfunction? PMID- 8730152 TI - Circulating endothelin-1 levels in systemic sclerosis subsets--a marker of fibrosis or vascular dysfunction? PMID- 8730153 TI - Sulfasalazine: a definitively efficient treatment for psoriatic arthritis. PMID- 8730154 TI - Raynaud's phenomenon as a presenting sign of ovarian adenocarcinoma. PMID- 8730155 TI - Polymyalgia rheumatica in patients with myelodysplastic syndromes. PMID- 8730156 TI - Effects of mirthful laughter on neuroendocrine and immune systems in patients with rheumatoid arthritis. PMID- 8730158 TI - Loss of contact sensitization evaluated by laser Doppler blood flowmetry and transepidermal water loss measurement. AB - Allergic contact dermatitis, which is mediated by activated T cells through a Type IV reaction, is usually believed to persist throughout life. There have been several reports of spontaneous loss of sensitization, e.g., loss of previous patch test reaction or loss of allergic contact dermatitis withal continuous exposure to the allergens. However, these have been well quantified. The aim of this study was to confirm the development of desensitization or hyposensitization with the aid of objective measurements. A total of 18 patients with alopecia areata, who received 4 months diphenylcyclopropenone (DPCP) contact immunotherapy, were patch tested and the responses were measured by laser Doppler blood flowmetry (LDF) and transepidermal water loss (TEWL). Patch tests were performed 2X, before and after 4 months of immunotherapy. To determine whether local desensitization developed, we compared change of DPCP concentrations producing mild contact dermatitis on the scalp. Our results showed that systemic and local hyposensitization occurred in human subjects by weekly applications of low concentrations of DPCP for 4 months. LDF seems to be more correlated with visual scores than TEWL measurement. PMID- 8730157 TI - Type II mixed cryoglobulinemia treated with fludarabine. PMID- 8730159 TI - High dissociation constants (pKa) of basic permeants are associated with in vivo skin irritation in man. AB - Previously, we have investigated the relationship between dissociation constant (pKa) and skin irritation potential. In the present experiment, 12 basic compounds, with varying pKa values ranging from 1.4 to 11.2, were applied on the backs of 12 healthy adult panellists. Cutaneous reactions were measured objectively, using reflectance spectroscopy and transepidermal water loss (TEWL), and evaluated subjectively with a modified Draize scale. A positive correlation between increasing pKa and skin irritation capacity, measured either visually or by reflectance spectroscopy, was found, but only mecamylamine (pKa = 11.2) induced a significant increase in transepidermal water loss. Compounds with low pKa also induced a paradoxical vasoconstriction measured by reflectance spectroscopy. Only high pKa appeared predictive of in vivo skin irritation, and these chemicals apparently induce skin irritation with only minimal disruption of the skin barrier. A simple 1-variable model is predictive of skin irritation for this series of organic permeants with increasing pKa. PMID- 8730160 TI - Development of hand eczema in junior hairdressers: an 8-year follow-up study. AB - The development of hand eczema was evaluated in an 8-year follow-up study carried out in 51 junior hairdressers. Predisposing factors were investigated. Data were collected by use of a self-administered questionnaire: 74% response. None of the junior hairdressers presented with hand eczema at the start of the survey. After 8 years, however, 51% had developed hand eczema. No significant difference in prevalence of hand eczema was found between practicing and non-practicing hairdressers (58% versus 33%). Development of hand eczema was not related to atopic constitution or nickel sensitivity. Dry skin type was associated with increased risk of developing hand eczema. PMID- 8730161 TI - Contact allergy to impurities in surfactants: amount, chemical structure and carrier effect in reactions to 3-dimethylaminopropylamine. AB - Since finding that all subjects with contact allergy to cocamidopropylbetaine give positive reactions to 3-dimethylaminopropylamine (DMPA), we wished to verify whether sensitization to other industrially-used tensioactives might also be due to content of DMPA as an impurity. We also investigated the possible "carrier action" that tensioactives might exert on minimal quantities of DMPA. Finally, we analyzed the relationship between the structure of DMPA and other chemically correlated molecules and their sensitizing potential, with particular reference to the structure of alkylamidopropylbetaines. For this purpose, in 34 patients with contact allergy to DMPA, we tested: (i) DMPA in concentrations below the threshold limit in water and in different tensioactives; (ii) substances that employ DMPA as a reagent in their synthesis; (iii) substances similar to DMPA as regards chemically reactive groups. The study showed that: (i) DMPA remains as a quantitatively detectable impurity in all tensioactives employing it in their synthesis; (ii) some common anionic (SLES) and non-ionic (polysorbate 20) tensioactives enhance the risk of sensitization from very low doses of DMPA, presumably due to a "carrier effect;" (iii) the sensitizing chemical structures in DMPA and related molecules are the primary amine and the tertiary (dimethyl substituted) amine groups, when separated by either 2 or 3 carbon atoms; (iv) no sensitizing action can be attributed to the functional groups present in alkylamidopropylbetaine molecules. PMID- 8730163 TI - Effect of glove occlusion on human skin (II). Long-term experimental exposure. AB - Gloves are indispensable in many occupations. Irritant skin reactions from gloves have, however, been reported. In the present study, the effect of long-term glove occlusion on normal skin (6 h/day for 14 days) was studied and, in addition, the effect of a cotton glove worn under the occlusive glove was also studied. 2 studies were performed (studies A and B). Study A: 19 volunteers wore an occlusive glove on normal skin 6 h/day for 14 days. They wore the glove on one hand only, while the other hand served as control. Study B: 18 volunteers wore occlusive gloves on both hands on normal skin. A cotton glove was worn under the occlusive glove on either the left or the right hand. Skin barrier function was evaluated by measurement of transepidermal water loss (TEWL) (Evaporimeter), skin hydration by electrical capacitance (Corneometer) and inflammation by erythema index (Derma-Spectrometer). The gloves used were hypoallergenic non-latex gloves. Results. Study A: glove occlusive on normal skin 6 h/day for 14 days had a significant negative effect on skin barrier function, as measured by TEWL. Study B: the negative effect on skin barrier function from occlusive gloves was prevented by the use of a cotton glove. It is concluded that gloves may be a substantial factor in the pathogenesis of cumulative irritant contact dermatitis, and that recommendations as to their use are important. PMID- 8730162 TI - Contact stomatitis due to palladium and platinum in dental alloys. AB - We report a patient with contact stomatitis due to combined sensitization to palladium and platinum. Patch testing showed strong and persistent allergic patch test reactions to palladium chloride (1% pet.), ammonium tetrachloroplatinate (0.25% pet.), and a palladium metal plate. A platinum metal plate showed a weaker reaction. Histological examination of a biopsy from the test site of palladium chloride (1% pet.) at D3 showed both eczematous and lichenoid changes. PMID- 8730164 TI - Oral mucosal diseases investigated by patch testing with a dental screening series. AB - The role of contact allergies in oral mucosal diseases was studied. The subjects were 24 patients out of 479 tested, who had oral mucosal symptoms and positive patch test reactions in a dental series during 1987-1994 at the Department of Dermatology, Helsinki University Hospital. The clinical diagnoses were oral lichen planus (LPO, 13 patients), leukoplakia (2), glossodynia, i.e., 'burning mouth syndrome' (4), stomatitis (3) and recurrent angioedema (2). Only 1 patient had symptoms in relation to dental care. All but 2 patients had allergic reactions to mercury (Hg) (12 patients), gold sodium thiosulfate (Au) (13 patients) or both. A clinical connection between oral symptoms and contact allergy was seen in 10 patients. 9 patients (7 LPO, 2 leukoplakia) had Hg allergy. In these cases, the oral lesions disappeared after the amalgam fillings had been removed. 1 patient had recurrent stomatitis and perioral eczema after dental care and 2,2-bis(4-(2-hydroxy-3-methacryloxypropoxy)phenyl)propane (BIS GMA) allergy. Her symptoms were caused by drilling of acrylic fillings. In addition, a connection between localized stomatitis and contact allergy was considered probable in 2 cases. 1 patient had stomatitis from contact with an orthodontic device and nickel allergy. The other had stomatitis from contact with a dental gold crown and gold allergy. No clinical connection was found between gold allergy and the oral symptoms of other patients. PMID- 8730166 TI - Ultrasound description and quantification of irritant reactions induced by dithranol at different concentrations. A comparison with visual assessment and colorimetric measurements. AB - Dithranol (D) is used as a therapeutic topical agent to treat psoriasis, although it produces inflammation and staining of skin and clothing. D-induced irritation has been evaluated by visual scoring and by bioengineering techniques, evidencing modifications of the inflammatory parameters, but no alterations of the skin barrier. The aim of our study was to evaluate the irritant reactions induced by D using ultrasound, and to compare the B-scanning data with visual assessments and colorimetric measurements. 13 healthy women underwent 2, 3-h patch tests with, respectively, 0.02% and 0.1% D in white petrolatum and 1 24-h patch test with 2% sodium lauryl sulfate (SLS). For assessing skin reactions, clinical judgement, colorimetry and echography were employed. Echographic images were evaluated by skin thickness measurements and segmentation procedures, using an 0-30 interval, marking the hyporeflecting areas in the dermis, and a 201-255 interval, assessing the hyperreflecting components of the image (both epidermal and dermal). In all subjects, D produced uniform reactions, the intensity varying according to concentration. Both echographic parameters of inflammation (skin thickness and 0 30 areas) showed an increase at all times of assessment. The 201-255D/0-30 pixel ratio, describing the distribution of dermal edema, indicated that strong inflammatory reactions, such as those caused by the application of 0.1% D, are accompanied by edema in the lower portion of the dermis. Finally, in contrast to SLS reactions, where a 24-h reduction in epidermal reflectivity was observable, D reactions appeared with an accentuation of the 201-255 epidermal band at 24-96-h examinations. PMID- 8730165 TI - Outcome of job change in patients with occupational chromate dermatitis. AB - Patients with allergic contact dermatitis due to dichromate are reputed to have a bad prognosis. Based on the Swiss Law on Accidents Insurance, the Swiss National Accidents Insurance Organization (SNAIO) may issue a declaration of medical inability (DMI) in cases of severe occupational dichromate dermatitis. With such a DMI, an employee is not allowed to perform any further work in contact with dichromate or cement. In this study, we reviewed medical records from 88 construction workers with DMI due to occupational dichromate dermatitis, between 1986 and 1989. Follow-up was performed by standardized questionnaire. 63 patients (72%) healed in the first few years after DMI. These patients mostly changed industry and strictly avoided all contact with cement or chromium salts. A few retired early. The outcome of our study is favorable in comparison to studies from other countries without the DMI mechanism. We conclude that strict allergen avoidance enforced by authorities, and financial support in the case of job change, are important factors in improving the prognosis in occupational dichromate dermatitis. PMID- 8730167 TI - Efficacy of 4 commercially available protective creams in the repetitive irritation test (RIT). AB - Protective creams (PCs) play their part in the prevention of occupational contact dermatitis, even though efficacy data are frequently lacking. 4 different commercially-available PCs were evaluated against a set of 4 standard irritants (10% sodium lauryl sulfate (SLS), 1% sodium hydroxide (NaOH), 30% lactic acid (LA) and undiluted toluene (TOL)) in the repetitive irritation test (RIT) in humans described by Frosch and Kurte. 20 subjects were tested on the paravertebral skin on the mid-back. Irritation was assessed by visual scoring, transepidermal water loss (TEWL) as an indicator of epidermal barrier function, and colorimetry as a parameter of inflammation. Very different protective effects of the PCs on irritation by chemical substances were detectable. All products were very effective against SLS irritation. No PC provided significant protection against toluene. 3 products showed a partially protective effect against all ionic irritants, while the 4th showed less protection against SLS and NaOH, and even amplification of inflammation by TOL. Considering the range of PC effects from good protection to increased irritation, depending on the irritant, the need for careful selection of PCs for specific workplaces is stressed. PMID- 8730168 TI - The occurrence and course of skin symptoms on the hands among female cleaners. AB - Skin symptoms on the hands were studied in questionnaire-based studies conducted in 1989 and 1991 among female cleaners employed at Danish nursing homes, schools and offices. A total of 1166 participated in 1989, and 1011 of them participated in 1991. The average age was 45 years and the average of seniority was 10 years. 1/5 of the cleaners reported problems with cleaning agents. A total of 81% had wet hands more than 1/4 of their working hours. A total of 43% reported having at least 1 out of 4 skin symptoms during a 1-year period. Among them, 70% reported improvement during weekends and holidays. A positive correlation was found between hours per week spent with wet hands and skin symptoms. During the 2-year follow-up period, the risk of developing skin symptoms was higher in the group that remained cleaners than in the group that left their cleaning jobs. Accordingly, the prognosis was better in the group that left their cleaning jobs compared to the group that remained cleaners. Retirement occurred more often among cleaners with skin symptoms than among the others. There is a future need to develop and implement new work organisation and cleaning methods to reduce the time spent with wet hands. PMID- 8730169 TI - Allergic contact dermatitis from cow hair and dander. PMID- 8730170 TI - Sensitization to benzocaine on a condom. PMID- 8730171 TI - Contact sensitization to organosilane solution in glass filament production. PMID- 8730172 TI - Investigation of the association between mercury sensitization and HLA-DR6. PMID- 8730173 TI - Contact urticaria caused by latex-free nitrile gloves. PMID- 8730174 TI - Contact urticaria from chironomids. PMID- 8730175 TI - Ampiroxicam-induced photosensitivity. PMID- 8730176 TI - Survey of airborne antibiotics. PMID- 8730177 TI - Occupational IgE-mediated protein contact dermatitis from pork in a slaughterman. PMID- 8730178 TI - Brain MR in Fukuyama congenital muscular dystrophy. AB - PURPOSE: To determine the MR characteristics of brain abnormalities in Fukuyama congenital muscular dystrophy (FCMD). METHODS: We reviewed 30 MR examinations of 21 patients with FCMD to assess cerebral and cerebellar cortical dysplasia, white matter changes, and miscellaneous abnormalities. RESULTS: On MR images, all patients had thick and bumpy cortices with shallow sulci corresponding to polymicrogyria, and 12 patients had pachygyric cortices with smooth surfaces, corresponding to type II lissencephaly. Both types of cortical dysplasia had characteristic distributions: the first type involved the frontal lobe in all 21 patients and also the parietotemporal lobe in 6 patients; the second type involved the temporooccipital lobes. Eighteen patients had prolonged T1 and T2 signal in the white matter, which was indistinct in neonates and seen infrequently in adolescents. In four patients, abnormal vessels were seen within the pachygyric cortices. CONCLUSION: MR studies of the brain show findings consistent with the known characteristics of FCMD. The MR detection of the two types of cerebral cortical dysplasia with characteristic distribution and cerebellar abnormalities is helpful in the differential and early diagnosis of FCMD. PMID- 8730179 TI - Imaging of the cobblestone lissencephalies. PMID- 8730180 TI - Skull base and calvarial deformities: association with intracranial changes in craniofacial syndromes. AB - PURPOSE: To analyze the skull and brain malformations in patients with craniofacial syndromes. METHODS: A retrospective analysis of imaging studies of 21 children with craniofacial anomalies (8 with Apert syndrome, 6 with Pfeiffer syndrome, 4 with Crouzon syndrome, 1 with Robert syndrome, 1 with Coffin-Lowry syndrome, and 1 with Saethre-Chotzen syndrome) was carried out using CT (21 patients), MR imaging (9 patients), and MR venography (2 patients). A series of qualitative and quantitative assessments of the skull base and intracranial structures was performed. RESULTS: Skull base abnormalities were present in all patients. Intracranial abnormalities included ventriculomegaly, frank hydrocephalus, callosal anomalies, hypoplasia/absence of the septum pellucidum, hypoplasia/dysplasia of the hippocampus, dysplasias or distortions of the cerebral cortex, and parenchymal hemorrhage. The anomalies of the corpus callosum, septum pellucidum, and hippocampus appeared primary, whereas the others may have been the result of brain distortion by the calvarial anomaly. MR imaging was more useful than CT for evaluating brain abnormalities. In the two patients in whom it was performed, MR venography showed anomalies of the venous system, indicating that venous anomalies, possibly related to the skull base hypoplasia, may contribute to the intracranial abnormalities. CONCLUSION: A wide range of neuroimaging abnormalities are present in the craniofacial syndromes. Some of these are clearly primary, whereas others appear to be related to the small skull base and sutural synostoses. MR venography may prove useful in defining the cause of some of the associated anomalies. PMID- 8730181 TI - Susceptibility-weighted MR for evaluation of vasodilatory capacity with acetazolamide challenge. AB - PURPOSE: To investigate cerebral vasodilatory capacity by acetazolamide challenge in healthy subjects and in patients with chronic occlusive cerebrovascular disease by using susceptibility-weighted gradient-echo MR imaging. METHODS: Eight patients with chronic occlusive cerebrovascular disease and four healthy volunteers were studied with susceptibility-weighted MR imaging before and after intravenous administration of 1000 mg of acetazolamide. Signal intensities were measured as a function of time in several regions of interest defined on anatomic images. In all patients with chronic occlusive cerebrovascular disease, acetazolamide challenge and resting regional cerebral blood flow were also evaluated with single-photon emission CT (SPECT). RESULTS: In healthy volunteers, signal intensities began to increase 3 to 4 minutes after acetazolamide administration, with a continuous increase during the subsequent 10 minutes. The effect lasted for approximately 45 minutes after administration. In patients with chronic occlusive cerebrovascular disease, signal changes on susceptibility weighted MR images of occluded areas with normal vasodilatory capacity on SPECT images did not differ from signal changes of nonocclusive areas. In those patients with changes that reflected diminished vasodilatory capacity, the MR images showed a lower percentage of signal changes after acetazolamide administration than those in normally perfused areas. CONCLUSION: Susceptibility weighted MR imaging offers an alternative method for estimating vasodilatory capacity. PMID- 8730182 TI - Quantitative high-resolution measurement of cerebrovascular physiology with slip ring CT. AB - PURPOSE: To implement and validate spiral slip-ring CT for use in cerebrovascular studies. METHODS: Continuous data were acquired from an experimental, first-pass, iodine contrast, bolus study by unidirectional X-ray tube rotation, and images were reconstructed at 100-millisecond intervals. Functional maps of cerebral blood volume (CBV) and cerebral blood flow (CBF) were constructed with voxel-by voxel gamma variate fitting. Reproducibility studies, different injection volumes and sites, and CO2 challenge were applied to verify the technique. RESULTS: Average absolute cortical gray and white matter and basal ganglia results were reproducible within +/- 0.8 ml/100 g for CBV and +/- 20 ml/100 g per minute for CBF, CBV response to changing arterial CO2 tension was significant only in cortical gray matter and basal ganglia; CBF response was significant in gray and white matter, as well as in the basal ganglia. CONCLUSION: Functional CT and constructed functional maps provide an optimal, high-resolution tool with which to visualize cerebrovascular parameters and their changes. PMID- 8730183 TI - Ipsilateral hemisphere activation during motor and sensory tasks. AB - PURPOSE: To compare activation of the ipsilateral cerebral hemisphere during tactile sensory and motor tasks involving the right and left hands. METHODS: Eight volunteers had functional MR imaging to measure the extent of cerebral hemisphere activation during a motor task and sensory task involving each hand. Hemispheric indexes (left hemisphere activation minus right hemisphere activation)/(left hemisphere activation plus right hemisphere activation) were computed for each hand and each task. The indexes for two tasks and the two hands were compared. RESULTS: The left-hand motor tasks activated the ipsilateral hemisphere in right handers significantly more than did the right-hand tasks. Motor tasks produced a greater activation of the ipsilateral hemisphere than did the sensory tasks. No significant differences were found between the hemispheric indexes for the right-hand and left-hand sensory tasks. CONCLUSION: This study confirms findings of a previous study, showing that the left hemisphere is active in left-hand motor tasks. Activation of the ipsilateral hemisphere is significantly less pronounced during sensory tasks than during motor tasks. PMID- 8730184 TI - Treatment of recurrent nasolacrimal duct obstructions with balloon-expandable metallic stents: results of early experience. AB - PURPOSE: To evaluate the efficacy of Palmaz balloon-expandable metallic stents for the maintenance of luminal patency in the treatment of recurrent nasolacrimal duct, obstructions after failed transluminal balloon dilatation. METHODS: Metallic stents were implanted in the nasolacrimal duct of four patients with recurrent epiphora. All the patients had already undergone transluminal balloon dilatation of the nasolacrimal duct with no or only temporary improvement. In two eyes, 9.5-mm-long Palmaz stents were used; in the other two eyes, 20-mm-long articulated-design Palmaz stents were placed under digital fluoroscopic monitoring. In each case, the upper tip protruded slightly into the lacrimal sac and the lower end was positioned inside the nasolacrimal duct. RESULTS: The stents were placed accurately in all cases, and no complications were observed. After stent placement, all patients had complete resolution of epiphora. During the follow-up period of 10 months, although complete obstruction did not occur in any case, complete resolution of epiphora was maintained only in one of four eyes. In two eyes, improvement was remarkable. In one eye with partial improvement, intrastent balloon dilatation was performed. CONCLUSION: Treatment of recurrent nasolacrimal duct obstructions with balloon-expandable metallic stents is a noninvasive, efficient, and safe outpatient procedure that may be an alternative to surgery as a means for managing epiphora. PMID- 8730185 TI - MR of optic papilla protrusion in patients with high intracranial pressure. AB - PURPOSE: To evaluate the signal characteristics of the optic papilla (optic nerve head) on routine cranial MR images in patients with clinical evidence of optic papilla elevation caused by high intracranial pressure, and to compare these findings with findings in healthy adult volunteers. METHODS: We reviewed retropectively the MR imaging examinations of 15 patients who were referred with objectively decreased visual acuity and funduscopic findings of optic papilla elevation. T1-weighted and T2-weighted axial MR images were obtained by using conventional spinecho acquisitions on 1.5-T MR imagers. In addition, the MR imaging studies in 10 healthy adult volunteers without visual impairment were reviewed as controls. RESULTS: In 10 (67%) of the 15 patients, visual elevation of the optic papilla was shown by MR imaging. In all 15 patients, the MR signal intensity of the optic papilla was hypointense relative to the vitreous of the globe on T2-weighted images. In the healthy volunteer group, the optic papillae were all similarly hypointense relative to the vitreous of the globe on T2 weighted images; however, these optic papillae were flat. CONCLUSION: Clinical examination and MR imaging may show elevation of the optic papilla in patients with high intracranial pressure. When chronic, optic papilla elevation has been shown to correlate well with severe loss of vision. Actual edema of the optic papilla seems to play little role in the physical elevation observed clinically in the chronic stages of this pathologic process. PMID- 8730186 TI - Relationship of the optic nerve to the posterior paranasal sinuses: a CT anatomic study. AB - PURPOSE: To delineate the relationship between the optic nerves and the posterior paranasal sinuses using CT data. METHODS: Direct coronal sinus CT scans of 150 consecutive patients with chronic inflammatory sinus disease were reviewed by two radiologists. Axial oblique reconstructions along the course of the optic nerve were obtained for the first 100 patients. The direct relationship between the optic nerve and the posterior ethmoid and sphenoidal sinuses was recorded, as were identations into the sinus wall, course of the nerve through the sinus region, pneumatization of the anterior clinoid process, and bone dehiscence. RESULTS: The relationship of the optic nerve to the posterior paranasal sinus fell into one of four discrete categories, type 1 through type 4. All 300 nerves were intimately related to the sphenoidal sinus. A small minority (3%) were in contact with the posterior ethmoidal sinus. Only type 4 nerves had contact with the posterior ethmoid air cell. Type 1 nerves course adjacent to the sphenoid sinus without indentation of the wall (228 nerves, 76%). Type 2 nerves course adjacent to the sphenoidal sinus, causing indentation of the sinus wall (44 nerves, 15%). Type 3 nerves course through the sphenoid sinus (19 nerves, 6%). Type 4 nerves course immediately adjacent to the sphenoidal sinus and the posterior ethmoidal air cell (9 nerves, 3%). Bone dehiscence over the optic nerve was found in 24% of the nerves; 4% of the optic nerves in our study had an associated pneumatized anterior clinoid process and 77% of these had an associated dehiscence over the optic canal. CONCLUSIONS: In all our cases the course of the optic nerve was adjacent to the sphenoidal sinus. Only 3% were in contact with the posterior ethmoidal sinus. Anatomic configurations that predispose the optic nerve to injury include type 2 or 3 optic nerves, bone dehiscence over the nerve, and pneumatization of the anterior clinoid process. These configurations are common and should be routinely sought out so that devastating complications from sinus surgery can be avoided. PMID- 8730187 TI - Arachnoid granulations in the transverse and sigmoid sinuses: CT, MR, and MR angiographic appearance of a normal anatomic variation. AB - PURPOSE: To investigate the imaging characteristics, prevalence, and clinical significance of arachnoid granulations in the transverse and sigmoid venous sinuses. METHODS: We reviewed the imaging findings, clinical signs and symptoms, final diagnoses, and follow-up studies of 32 patients with 41 probable arachnoid granulations. RESULTS: On CT scans, arachnoid granulations appear as well-defined filling defects, wholly or partly within a venous sinus, with the same density as cerebrospinal fluid. MR images show these entities as largely isointense with cerebrospinal fluid in all sequences. Linear variations of signal intensity within the granulations are thought to be fibrous septa or vessels. Calcification was present in 3 granulations and altered both CT density and MR signal intensity. The granulations appear as filling defects at MR angiography and at digital subtraction angiography. In some oblique MR angiographic projections, they appear elliptical and could be mistaken for thrombus. No clinical significance could be given to the existence of any of these arachnoid granulations. They occur in 0.3 to 1 of 100 adults in the population. CONCLUSION: Arachnoid granulations in the transverse and sigmoid venous sinuses are common findings seen with thin-section imaging and are usually of no significance. PMID- 8730188 TI - The effect of flow arrest on distal embolic events during arterial occlusion with detachable coils: a canine study. AB - PURPOSE: To determine the effect of proximal flow arrest on the frequency and timing of distal embolic events during occlusion of the common femoral artery with detachable coils. METHODS: Twenty-three complex fibered platinum coils were delivered into 10 common femoral arteries without proximal flow arrest. The arteries were continuously monitored for flow and embolic events by Doppler sonography during delivery and for at least 10 minutes after delivery of each coil. Thirty-four coils were delivered into 6 arteries after proximal flow arrest by inflation of a nondetachable balloon. After balloon deflation, each artery was monitored by Doppler sonography for 10 minutes. RESULTS: In the 10 arteries occluded without flow arrest, 87 events (8.7 per artery) occurred, of which 47 were embolic and 40 were indeterminate. In the 6 arteries with flow arrest, the number of emboli detected was 3 (0.5 per artery). Embolic events occurred only if there was residual flow. In those arteries that were occluded when the flow arrest balloon was deflated, no emboli were detected. CONCLUSIONS: Proximal flow arrest virtually eliminates the risk of distal emboli during arterial occlusion with detachable fibered coils. The use of fibered coils, in conjunction with proximal flow arrest, allows for safe arterial occlusion when detachable balloons are not available or their use is not feasible. PMID- 8730189 TI - Use of intravascular stents in the treatment of internal carotid and extracranial vertebral artery pseudoaneurysms. AB - The management of extracranial carotid or vertebral artery pseudoaneurysms is controversial. Although some of these lesions resolve spontaneously, many clinicians opt to treat them with trapping procedures that result in vessel sacrifice. We describe two cases in which an intravascular stent was used to obliterate an aneurysm of the extracranial vertebral artery and the internal carotid artery, respectively, while maintaining the patency of the parent vessel. The technique, which has been successful in experimental animal models, shows promise for application in humans. PMID- 8730190 TI - Use of large-caliber coronary guiding catheters for neurointerventional applications. AB - The use of certain large-caliber (ie, 7F to 9F) coronary guiding catheters for selected neuroendovascular procedures is described in the context of surveying recently developed neurovascular guiding catheters. These large-caliber guiding catheters have proved useful in situations requiring proximal access to tortuous brachiocephalic arteries, permitting coaxial delivery of relatively large neurointerventional devices, such as most detachable balloons and 5F percutaneous transluminal angioplasty catheters. These large-caliber coronary guiding catheters have some important advantages over both old and new neurovascular guiding catheters, including favorably preshaped configurations and superior torque control. These features permit relatively easy direct selective catheterization of unfavorable vascular anatomy resulting from dolichoectasia of the aortic arch and great vessels. PMID- 8730191 TI - Cervical spondylosis: three-dimensional gradient-echo MR with magnetization transfer. AB - PURPOSE: To compare a three-dimensional Fourier transform (3DFT) gradient-echo pulse sequence with magnetization transfer at a short echo time against standard 3DFT gradient-echo technique in the evaluation of cervical spondylosis, specifically addressing the effects of motion and susceptibility artifacts on the dimensions of the neural foramina and contrast at the cerebrospinal fluid (CSF) spinal cord interface. METHODS: Ten patients with clinically suspected cervical spondylosis were examined with axial MR imaging using both our standard 3DFT gradient-echo sequence and a 3DFT gradient-echo sequence with a short echo time plus magnetization transfer. Two neuroradiologists measured the transverse dimensions of 22 diseased neural foramina and graded the contrast at the CSF spinal cord interface. RESULTS: Sixteen of 22 affected neural foramina were larger in the transverse dimension when the magnetization transfer technique was used than when the standard 3DFT gradient-echo sequence was used. In 9 of 10 patients superior contrast was seen at the CSF-spinal cord interface on images obtained with the magnetization transfer technique. CONCLUSIONS: In the cervical spine, 3DFT gradient-echo imaging with magnetization transfer improves contrast and sharpness of the CSF-spinal cord interface at short echo times. This results in less exaggeration of the neural foraminal stenosis as compared with that seen with standard 3DFT gradient-echo techniques owing to the diminished effects of motion and susceptibility artifacts. PMID- 8730192 TI - Spinal angiolipomas: MR features. AB - PURPOSE: To determine the MR features of spinal angiolipomas and to compare these findings with their histologic appearance. METHODS: The MR examinations of three patients with surgically proved angiolipomas were reviewed for tumor location and extent, signal characteristics, and pattern of contrast enhancement, and were then compared with the histologic findings. RESULTS: Four tumors were found in the three patients, all located in the posterior epidural compartment, averaging about 2.5 vertebral bodies in length. On noncontrast T1-weighted images, all lesions were inhomogeneous and hypointense relative to epidural fat. Inhomogeneous enhancement was seen in three lesions on postcontrast T1-weighted images obtained with fat-saturation techniques. Angiolipomas were least conspicuous on T2-weighted images. A high vascular content correlated with the presence of large hypointense regions on T1-weighted images. CONCLUSION: Spinal angiolipomas are typically hyperintense on noncontrast T-1-weighted images relative to other tumors. Angiolipomas that contain large hypointense foci on noncontrast T1-weighted images can be expected to have a high degree of vascularity. PMID- 8730193 TI - A history of neuroradiology in Italy. PMID- 8730194 TI - Brain death: MR and MR angiography. AB - SUMMARY: Four patients in whom brain deaths was identified on the basis of neurologic and electroencephalographic findings were examined with MR imaging and MR angiography. MR images showed diffuse swelling of the cerebral gyri and cerebellar cortex, which prolongation of both the T1 and T2 signal (representing hypoxic ischemic brain injury), downward displacement of the diencephalon and the brain stem (central and tonsillar herniation), and loss of flow void in the intracranial portions of both internal carotid arteries. MR angiograms did not show the intracranial vessels above the level of the supraclinoid portion of the internal carotid arteries. MR angiography and MR imaging are noninvasive and reliable methods for use in determining brain death. PMID- 8730195 TI - Noninvasive evaluation of malignancy of brain tumors with proton MR spectroscopy. AB - PURPOSE: To test clinical proton MR spectroscopy as a noninvasive method for predicting tumor malignancy. METHODS: Water-suppressed single-voxel point resolved spectroscopy in the frontal white matter of 17 healthy volunteers and 25 patients with brain tumors yielded spectra with peaks of N-acetyl aspartate (NAA), choline-containing compounds (Cho), creatine/phosphocreatine (Cre), and lactate. These peak intensities were semiquantitated as a ratio to that of the external reference. The validity of the semiquantitation was first evaluated through phantom and volunteer experiments. RESULTS: The variation in measurements of the designated region in the volunteers was less than 10%. Normal ranges of NAA/reference, Cho/reference, and Cre/reference were 3.59 +/- 0.68, 1.96 +/- 0.66, and 1.53 +/- 0.64 (mean +/- SD), respectively. In 17 gliomas, the Cho/reference value in high-grade gliomas was significantly higher than in low grade gliomas. Levels of NAA/reference were also significantly different in low grade and high-grade malignancy. In eight meningiomas (four newly diagnosed and four recurrent), the level of Cho/reference was significantly higher in recurrent meningiomas than in normal white matter or in newly diagnosed meningiomas. CONCLUSIONS: Higher grades of brain tumors in this study were associated with higher Cho/reference and lower NAA/reference values. These results suggest that clinical proton MR spectroscopy may help predict tumor malignancy. PMID- 8730197 TI - Evolution of high-intensity basal ganglia lesions on T1-weighted MR in neurofibromatosis type 1. AB - PURPOSE: To characterize the temporal evolution of the foci of T1 shortening in basal ganglia lesions in patients with neurofibromatosis type 1 (NF-1). METHODS: A retrospective review of MR images of 37 patients with NF-1 revealed 8 patients in whom regions of T1 shortening were noted in the basal ganglia. We reviewed sequential images obtained in these selected patients with special attention to chronological changes in the foci of T1 shortening and their relationship to changes on T2-weighted images. RESULTS: Regions of short T1 in the globus pallidus were observed in 8 patients. In 2 of 3 patients in whom foci of T1 shortening were not identified on the initial imaging study, T1 shortening developed and T2 prolongation diminished after an initial increase. In the third patient, T1 and T2 prolongation appeared simultaneously. Sequential scans in the other 5 patients, in whom areas of increased signal intensity in the globus pallidus were present on both T1-weighted and T2-weighted images on the initial MR examination, showed a diminution in the size of the region of T2 prolongation in 2 patients, an increase in the size of the region of T2 prolongation in 1 patient, a mixed pattern of change in the size of the region of T2 prolongation in 1 patient, and no change in the region of T2 prolongation in 1 patient. During the periods of these T2 changes, the areas of T1 shortening showed no significant interval change. CONCLUSION: The foci of prolonged T2 relaxation in the basal ganglia appear to evolve in a manner similar to the foci of T2 prolongation in the white matter of the posterior fossa. However, the corresponding foci of short T1 in the basal ganglia may evolve with a different time course. In some patients, the foci of short T1 develop at a later time than the T2 prolongation and progress; these foci of short T1 do not appear to regress over periods as long as 90 months. Possible causes of the T1 shortening are remyelination and calcification. PMID- 8730196 TI - Abnormal ocular enhancement in Sturge-Weber syndrome: correlation of ocular MR and CT findings with clinical and intracranial imaging findings. AB - PURPOSE: To estimate the prevalence of abnormal ocular enhancement in children with Sturge-Weber syndrome as detected with MR imaging and CT and to correlate this with the clinical, fundoscopic, and intracranial imaging findings. METHODS: Fifteen children, 4 years old or younger, with Sturge-Weber syndrome were examined with enhanced CT and MR imaging. Eleven children had unilateral intracranial involvement and 4 had bilateral involvement, for a total of 19 abnormal hemispheres and related orbits. The presence of ocular enhancement was compared with the fundoscopic findings independently. Ocular enhancement was correlated with the extent of leptomeningeal disease, the severity of the cutaneous lesion, and the presence of glaucoma by the calculation of likelihood ratios and 95% confidence limits. RESULTS: Seven of the 15 patients had abnormal ocular enhancement, which was present in 10 (53%) of the eyes associated with the 19 abnormal hemispheres. MR imaging showed choroidal hemangioma in 7 of 8 patients in whom hemangiomas were shown at fundoscopy. The likelihood of ocular enhancement was increased with the presence of bilateral disease, extensive facial nevi, and glaucoma; there was no significant correlation with the extent of hemispheric involvement. CONCLUSION: Both enhanced MR imaging and CT can show diffuse choroidal hemangioma in patients with Sturge-Weber syndrome. However, MR imaging is more sensitive and is recommended to aid in the detection of abnormalities with preventable late complications. PMID- 8730198 TI - Subacute sclerosing panencephalitis: evaluation with CT and MR. AB - PURPOSE: To evaluate the progression of CT and MR changes of the brain in subacute sclerosing panencephalitis (SSPE) as a basis for assessing the effects of different types of therapy. METHODS: Fifty-two patients with SSPE were examined, 44 with MR imaging and 42 with CT of the brain on one or more occasions. A total of 92 MR and 67 CT studies were performed. RESULTS: Correlation between the clinical status and the MR findings in admission was poor. Of 20 patients with clinically advanced disease, only 8 had marked MR abnormalities; 6 had normal or almost normal findings on MR examinations. Two of 4 patients with clinically mild disease had advanced MR changes. The progression of the MR findings appeared to follow a constant pattern. The earliest pathologic finding was focal, high-T2-intensity white matter changes; later atrophic changes followed. The atrophy lagged behind the white matter changes and was thus mild when white matter changes were moderate or severe. In the most advanced stage, when the patient was in a neurovegetative state, an almost total loss of white matter had usually taken place. At this stage, the corpus callosum was also thin. Basal ganglia changes, usually involving the putamina, were seen in one third of patients and cortical gray matter changes were seen in one fourth of patients examined with MR imaging. In 2 of 20 patients, MR changes regressed in parallel with clinical improvement following therapy, but in 5 patients clinical improvement was accompanied by progression of MR changes. CONCLUSION: The progress of MR abnormalities seen in patients with SSPE seems to follow a constant pattern, but the severity of MR changes does not always correlate well with the clinical findings. Caution must therefore be used when evaluating the effects of therapy. PMID- 8730199 TI - Contrast-enhanced magnetization transfer saturation imaging improves MR detection of herpes simplex encephalitis. AB - Cranial MR imaging was performed in three patients in whom herpes simplex encephalitis was subsequently proved. In all cases, the postcontrast T1 weighted MR images obtained with magnetization transfer saturation showed greater central nervous system involvement than was apparent on the conventional MR images. Specifically, the postcontrast magnetization transfer images were superior at delineating generalized meningeal enhancement as well as focal areas of brain involvement not seen on noncontrast T2-weighted images orconventionalpostcontrast T1-weightedimages. PMID- 8730200 TI - Transcranial color-coded duplex sonography in unilateral flow-restrictive extracranial carotid artery disease. AB - PURPOSE: To provide transcranial color-coded duplex flow-velocity data for the basal cerebral arteries in patients with unilateral flow-restrictive extracranial carotid artery disease, and to compare these data with the flow velocities obtained in healthy control subjects. METHODS: Transcranial color-coded duplex sonography was performed in 78 patients with different patterns of cross flow through the anterior and posterior communicating arteries associated with unilateral obstruction (70% to 100%; 46 stenoses and 32 occlusions) of the internal carotid arteries. Peak systolic, mean, and end diastolic velocities were measured in the anterior, middle, and precommunicating and postcommunicating posterior cerebral arteries. These measurements were compared with the values obtained in 125 age- and sex-matched health control subjects. RESULTS: Patients with anterior communicating artery cross flow to the middle cerebral artery (63%) had increased peak velocity in the anterior cerebral artery and decrease peak velocity in the middle cerebral artery on the obstructed (ipsilateral) side, and increased peak velocity in the anterior cerebral artery on unobstructed (contralateral) side. Patients with anterior communicating artery cross flow to the pericallosal artery (19%) had increased contralateral peak systolic velocity and mean anterior cerebral artery velocities. Patients without anterior communicating artery cross flow (18%) had normal peak velocities in the anterior and middle cerebral arteries. Patients with posterior communicating artery cross flow (42%) had ipsilaterally decreased peak systolic and mean middle cerebral artery velocities and increased peak velocities in the precommunicating posterior cerebral artery. Patients without posterior communicating artery cross flow (58%) had ipsilaterally decreased peak systolic and mean middle cerebral artery velocities. CONCLUSION: Our findings suggest that typical abnormalities of basal cerebral artery flow velocities occur in patients with unilateral 70% to 100% obstruction of the internal carotid arteries resulting in different patterns of cross flow through the circle of Willis. PMID- 8730201 TI - Wegener granulomatosis: CT and MR findings. AB - PURPOSE: To demonstrate the spectrum of CT and MR imaging findings in patients with Wegener granulomatosis and to determine how often these findings could be attributed to either direct extension from paranasal or orbital disease sites, remote granulomas, or central nervous system (CNS) vasculitis. METHODS: We retrospectively reviewed the CT or MR studies of 15 patients with Wegener granulomatosis. RESULTS: Abnormal findings were seen in 7 patients (5 examined with MR imaging, 2 with CT). Findings included dural thickening and contrast enhancement (3 patients), infarcts (2 patients), regions of hyperintense signal on T2-weighted MR images (2 patients), and abnormal MR signal in the brain stem (2 patients). Three patients with imaging findings of dural enhancement and thickening were thought to have remote granulomatous lesions involving the dura. No patients had extension from sites external to the CNS or clinical findings suggestive of CNS vasculitis. CONCLUSION: The spectrum of CT and MR findings in Wegener granulomatosis includes dural thickening and enhancements cerebral infarction, and MR signal abnormalities in the brain stem and white matter. Presumed remote granulomatous lesions were the most common causes of CNS findings in this study. Complications related to non-CNS disease (eg, hypertension, endocarditis) also appear to have played a role in some patients. PMID- 8730202 TI - Accuracy of coregistration of single-photon emission CT with MR via a brain surface matching technique. AB - We describe a technique of brain surface matching of single-photon emission CT and MR images in human subjects and document the accuracy of this technique with the use of fiduciary markers. This mismatch averaged 4.3 mm as measured by the fiduciary markers and 2.1 mm as measured by the root mean square distance. PMID- 8730203 TI - MR appearance of cerebral Drancunculus borealis infection. PMID- 8730204 TI - MR signal of mamillary bodies. PMID- 8730205 TI - Widening of Virchow-Robin spaces. PMID- 8730206 TI - Is high signal intensity in the corticospinal tract a sign of degeneration? PMID- 8730207 TI - N-methyl-D-aspartate-stimulated increases in intracellular calcium exhibit brain regional differences in sensitivity to inhibition by ethanol. AB - This study compared N-methyl-D-aspartate (NMDA)-stimulated increases in intracellular calcium in fura-2-loaded neurons dissociated from newborn rat brainstem (EC50 in microM; 6.4), cerebellum (9.5), forebrain (6.3), and hippocampus (10.6). Ethanol inhibition of the response to 25 microM NMDA differed among the regions. The NMDA response in hippocampus was inhibited by 20 mM ethanol; cortex and cerebellum responses were inhibited by 80 mM ethanol, and no inhibition was seen in the brainstem. Addition of glycine (15 microM) failed to attenuate ethanol inhibition of the NMDA response. These results demonstrate that ethanol inhibition of NMDA-stimulated responses varies according to brain region. In contrast to previous findings from this laboratory using dissociated neurons from whole brain, the addition of glycine did not reverse the inhibitory effects of ethanol on NMDA-stimulated responses. PMID- 8730209 TI - Induction of cytochrome P-4502E1 by ethanol in rat Kupffer cells. AB - Ethanol has been shown to affect several Kupffer cell functions, but the mechanisms underlying these changes are unknown. One possible mediator is cytochrome P-4502E1 (CYP2E1), an ethanol-inducible enzyme that has been associated with toxic effects in the liver, as well as in many extrahepatic organs. To assess whether CYP2E1 can be induced by ethanol in Kupffer cells, male rats pair-fed ethanol-containing or control Lieber-DeCarli diets for 3 weeks were studied. Immunoblotting experiments showed that ethanol-treatment caused a 7-fold increase in CYP2E1 content both in Kupffer cells and hepatocytes. When expressed per milligram of S9 protein, the content of CYP2E1 in Kupffer cells was, however, 10 times lower than in hepatocytes. Immunohistochemical studies revealed that CYP2E1 is located in the endoplasmic reticulum of Kupffer cells in vivo and that it is also present in isolated Kupffer cells. In both Kupffer cells and hepatocytes, ethanol feeding increased the hydroxylation of p-nitrophenol, a relatively specific substrate for CYP2E1, demonstrating that the induced CYP2E1 was catalytically active. This reaction was significantly inhibited by anti CYP2E1 IgG in both types of cells. Although CYP2E1 may not be the predominant pathway for ethanol metabolism in hepatocytes, it is possibly the major one in Kupffer cells. Thus, the induction of CYP2E1 by ethanol in these cells could cause significant changes in intracellular acetaldehyde concentrations which, together with increased lipid peroxidation, may contribute to the development of alcoholic liver injury. PMID- 8730208 TI - Ethanol consumption and taste preferences in C57BL/6ByJ and 129/J mice. AB - Mice of the C57BL/6ByJ (B6) and 129/J (129) strains were offered different concentrations of taste solutions in 48-hr, two-bottle choice tests. In comparison with the 129 strain, the B6 strain had higher preferences for ethanol, sucrose, and citric acid. They had lower preferences for NaCl and similar preferences for capsaicin and quinine hydrochloride. These data are consistent with the hypothesis that the higher ethanol intake by B6 mice depends, in part, on higher hedonic attractiveness of its sweet taste component. PMID- 8730210 TI - Isolation and characterization of three alcohol dehydrogenase isozymes from Syrian golden hamsters. AB - Electrophoresis of freshly prepared tissue homogenates of the Syrian golden hamster (Mesocricetus auratus) on starch gel followed by activity staining with ethanol as the substrate revealed three major alcohol dehydrogenase (ADH) isozymes. One of these isozymes, TT-ADH, found only in the testes of golden hamsters was previously purified and partially characterized (Keung WM: Biochem. Biophys. Res. Commun. 156:38-45, 1988). The other two, AA- and BB-ADH, which are most abundant in the liver, have now been purified by affinity chromatography on 4-(3-(N-(6-aminocaproyl)amino)propyl)pyrazole-sepharose and testosterone-17 beta hemisuccinate-agarose. Hamster AA-, BB-, and TT-ADH are all homodimers of molecular weight near 80,000 and each contains 4 atoms of zinc. Amino acid analyses show that BB-ADH is most closely related to the gamma-form of human class I ADH, whereas AA- and TT-ADH are most closely related to the beta-form of the human enzyme. BB-ADH is the only hamster ADH that is active toward sterols and sensitive to testosterone and isoflavone inhibition. These results suggest that hamster BB- and human gamma gamma-ADH also share similar catalytic properties. AA- and TT-ADH are neither active toward sterols nor sensitive to testosterone or isoflavone inhibition; thus, they are functionally different from the human alpha alpha- or gamma gamma-ADHs. Compared with AA- and BB-ADH, TT-ADH exhibits much higher Km values toward primary aliphatic alcohols and cyclohexanol. AA- and BB-ADH share similar substrate specificities toward primary aliphatic alcohols. However, they exhibit different stereospecificities for secondary alcohols. BB-ADH prefers the (R)-(-)-isomer of 2-butanol, whereas AA ADH prefers the (S)-(-)-isomer. These results further demonstrate that catalytically, hamster BB- and AA-ADH belong to different subfamilies of class I ADH. PMID- 8730211 TI - Suppression of alcohol intake after administration of the Chinese herbal medicine, NPI-028, and its derivatives. AB - The Chinese herbal medicine, NPI-028, has been used for centuries in China to counteract alcohol intoxication. The present study used a number of different experimental conditions to determine whether NPI-028 and its derivatives might selectively influence alcohol intake in rodents that naturally exhibit high alcohol intakes. It was determined that intraperitoneal (i.p.) injections of NPI 028 (0.5, 0.75, and 1.0 g/kg) suppressed alcohol intake by up to 30% in both alcohol-preferring P and Fawn-Hooded (FH) rats during a continuous access schedule. These injections did not significantly affect food or water intakes, nor did the highest dose of NPI-028 (1 g/kg) alter blood ethanol levels after an i.p. injection of 2.5 g/kg of ethanol. In P rats, it was found that NPI-028 was orally active with the dose of 1.5 g/kg having a greater effect on ethanol intake than the 1.0 g/kg dose; once again, food and water intakes were not significantly altered. In FH rats maintained on a limited access schedule (1 hr/day), alcohol intake was completely abolished by 1.5 g/kg of NPI-028. Chronic i.p. administration of NPI-028 (0.75 g/kg) for four consecutive days in FH rats maintained on a continuous access schedule did not lead to any diminution of its alcohol-suppressant effects. Thus, NPI-028 has significant effects on alcohol intake without much effect on water and food intake, and tolerance does not readily develop to these effects. The i.p. administration of a partially purified extract (NPI-031) of NPI-028, obtained by countercurrent chromatography, also dose-dependently suppressed ethanol intake in FH rats, but the highest dose 200 mg/kg) also significantly decreased food intake. Finally, the i.p. administration of puerarin (NPI-31G), an isoflavone isolated from NPI-031 by countercurrent chromatography, significantly reduced ethanol intake in FH rats without affecting food or water intake. Therefore, NPI-028 and one of its pure components, NPI 031G, selectively reduced ethanol intake in alcohol-preferring rats. PMID- 8730212 TI - Association of alcoholism with the N-glycosylation polymorphism of pseudodeficient human arylsulfatase A. AB - The IIIa and IIIb electrophoretic variants of arylsulfatase A (EC 3.1.6.8) are 12 times more prevalent in alcoholic than in nonalcoholic populations. These variant enzymes, found in a subset of alcoholics, possess the pseudodeficient Asn350-Ser mutation of arylsulfatase A and, consequently, lack an N-linked glycan unit. These genetically determined variants of arylsulfatase A show reduced intracellular half-life, and cells from such individuals possess reduced enzymic activity. We propose that this polymorphism is an underlying genetic and biochemical factor contributing to the neuropathology and/or addiction pathway of this disease. PMID- 8730213 TI - Structural characterization of variant forms of arylsulfatase A that associate with alcoholism. AB - Several electrophoretic forms of human platelet arylsulfatase A (ASA), including variant type IIIa and normal type IV(a), have been identified by nondenaturing polyacrylamide gel electrophoresis. An alcoholic population that we have analyzed is enriched in variant type IIIa compared with nonalcoholic psychiatric and normal controls. Individuals with the IIIa enzyme possess greatly reduced levels of ASA activity. To understand further the structural basis for the differences and their potential biological consequences, the nature of the ASA variant expressed by fibroblasts from different individuals was explored. The electrophoretic patterns of fibroblast ASA from the IIIa and IV(a) individuals differ in degree of phosphorylation. Furthermore, fibroblast ASA from IIIa individuals lacks an N-linked glycan found in ASA from IV(a) individuals. In addition, differences in peptide and/or posttranslational modification unrelated to the N-linked carbohydrate or phosphorylation exist between the fibroblast ASA from IIIa and IV(a) individuals. The finding that both fibroblasts and platelets exhibit related electrophoretic isoform patterns characteristic of the donor's ASA type allows for the use of fibroblasts to study the impact of ethanol on the metabolism of cells possessing different ASA types. PMID- 8730214 TI - Alcohol stimulates ACTH secretion in the rat: mechanisms of action and interactions with other stimuli. AB - This review discusses some of the mechanisms through which alcohol (EtOH) alters the activity of the hypothalamic-pituitary-adrenal (HPA) axis. In adult rats, acute EtOH treatment increases plasma ACTH and corticosteroids levels primarily by stimulating the release of corticotropin-releasing factor (CRF) and possibly vasopressin (VP) from nerve terminals in the median eminence. Increased CRF gene transcription in the hypothalamus may also be important. The HPA axis remains activated during chronic EtOH exposure, although habituation may take place. Changes in the responsiveness of hypothalamic neurons, a phenomenon itself dependent in part on a number of intermediate secretagogues, as well as decreased pituitary responsiveness to VP, all play a role. Finally, the activity of the HPA axis is influenced by exposure to EtOH during embryonic development, with mature offspring showing hyporesponsiveness to many stimuli. These altered responses appear to be caused in part by changes in the synthesis/release CRF, possibly under the influence of nitric oxide. CRF, VP, ACTH, and corticosteroids are important regulators of the immune system, behavior, metabolic pathways, and reproductive parameters. Alcohol therefore may influence such functions through the pathological secretion of these hormones. A better understanding of the mechanisms through which the drug alters their release thus may permit the development of therapies designed to alleviate some of the consequences of alcoholism. PMID- 8730216 TI - Repeated ethanol withdrawal produces site-dependent increases in EEG spiking. AB - Recent studies have suggested an important kindling-like exacerbation of ethanol withdrawal symptoms after repeated cycles of ethanol intoxication and withdrawal. Few studies, however, have evaluated the effect of multiple episodes of intoxication and withdrawal on spontaneous EEG activity after cessation of ethanol intake. In this study, electrographic activity in cortical and subcortical structures male Sprague-Dawley rats was examined after multiple cycles of ethanol intoxication and withdrawal. After surgical implantation of electrodes, animals received repeated cycles of chronic ethanol exposure in vapor inhalation chambers for 10 or 20 days, with 4-day withdrawal periods between each. Upon removal from the inhalation chamber, spontaneous EEG activity was recorded intermittently for 72 hr. These data were then examined for the presence of spikes and sharp waves. Results indicate that the levels of spike and sharp wave activity observed vary with both length of ethanol exposure and with the number of withdrawal cycles, and that these effects varied with neuronal site. Changes in spike and sharp wave activity were first observed within hippocampal areas, with other subcortical and cortical sites showing increased activity after additional ethanol exposure or additional cycles of intoxication and withdrawal. Hippocampal areas CA1 and CA3 differed significantly from one another in their response to chronic ethanol exposure, with area CA1 most affected by changes in amount of ethanol exposure and are CA3 most affected by number of withdrawal cycles. These results indicate an increased severity of the ethanol withdrawal syndrome after repeated ethanol withdrawal episodes and suggest differential, site-specific changes in neuronal excitability. PMID- 8730215 TI - Gender difference in alcohol-evoked hypothalamic-pituitary-adrenal activity in the rat: ontogeny and role of neonatal steroids. AB - Alcohol administration results in activation of the hypothalamic-pituitary adrenal (HPA) axis, with female rats secreting more adrenocorticotropin (ACTH) and corticosterone (B) than males in response to the same dose of alcohol. We first examined the ontogeny of the gender difference in HPA responsiveness to alcohol by administering four doses (0, 1, 2, or 3 g/kg body weight) to animals at 21, 41, and 61 days of age (prepubertal, peripubertal, and postpubertal, respectively). We then investigated the organizational role of steroids by manipulating the neonatal steroidal milieu. Rats of both genders were gonadectomized or injected with testosterone propionate within 24 hr of birth and the HPA response to 3 g/kg body weight alcohol was tested in adulthood (postpubertal period). Our data show that the gender difference in HPA responsiveness to alcohol administration arises peripubertally. In addition, HPA response to alcohol is quantitatively smaller in intact male rats than in feminized groups (gonadectomized males and females, intact females) and masculinized female rats. We conclude that the gender difference in HPA response to alcohol observed in postpubertal rats injected with alcohol depends on the activational role of testicular androgens, rather than on their organizational influence. PMID- 8730217 TI - Reduced fatty acid ethyl ester synthase activity in the white blood cells of alcoholics. AB - PURPOSE: Fatty acid ethyl esters (FAEEs), esterification products of ethanol and fatty acids, have been implicated as mediators of ethanol-induced organ damage. It has been shown that FAEE synthase, the enzyme responsible for the formation of FAEE, is present selectively in the organs commonly damaged by ethanol abuse. Recently, we have made the observation that FAEEs are also present in the serum after ethanol ingestion. The current study was performed to determine whether cellular elements of the blood and/or plasma are capable of synthesizing FAEEs from fatty acids and ethanol. MATERIALS AND METHODS: Heparinized blood samples were collected from 10 healthy volunteers, and the red blood cells, platelets, plasma, and several white blood cell populations were assayed for FAEE synthase activity. Blood samples from control subjects and individuals admitted to an alcoholic detoxification unit at a local hospital were also assayed for FAEE synthase activity. RESULTS: We observed that the FAEE synthase activity is present in whole blood, primarily within white blood cells. Fractionation of the white blood cells revealed that the lymphocyte-monocyte fraction isolated using Ficoll-hypaque contained approximately 3.5-fold higher activity than the granulocyte fraction. The cell type that contained the highest FAEE synthase activity (1220 pmol/hr/10(6) cells) was the natural killer (NK) cell population. B cells contained approximately 40% of the enzyme activity found in NK cells, and the B-cell activity was slightly greater than that found in CD4+ and CD8+ T cells. Having shown that FAEE synthase exists in a blood cell, we subsequently demonstrated that alcoholic individuals have approximately half the white blood cell FAEE synthase activity of that found in normal controls. We also demonstrated that white blood cell FAEE synthase could be induced nearly 2-fold upon ingestion of 2 oz of scotch whiskey for 6 days. The enzyme activity returned to baseline levels despite ingestion of 2 oz of scotch whiskey/day for 3 additional days. CONCLUSIONS: These data indicate that ethanol ingestion results in increased FAEE production, particularly by NK cells. FAEE synthesis after ethanol ingestion may explain the presence of FAEE in the serum. The lower enzyme activity observed in white blood cells of alcoholics from a detoxification center may be the result of years of ethanol abuse or it may be that alcoholics congenitally have low levels of FAEE synthase. If the latter is true, this finding may explain in part the genetic predisposition of many alcoholic individuals to ethanol abuse. PMID- 8730218 TI - Human neutrophil functions are inhibited in vitro by clinically relevant ethanol concentrations. AB - Neutrophils [polymorphonuclear neutrophils (PMNs)] play a pivotal role in host defense in man. These defenses may be compromised, however, in alcohol users and abusers. We therefore evaluated the effect of ethanol levels (12.5 to 500 mg/dl), on key functions of human PMNs-chemotaxis and production of reactive oxygen species-and on changes in cytosolic-free calcium ([Ca2+]i), a pivotal intracellular mechanism of PMN activation. Ethanol significantly inhibited chemotaxis as evaluated by formyl-methionyl-leucyl-phenylalanine (fMLP)-induced upregulation of surface adhesion molecules (CD11b). fMLP-induced PMN elongation was only inhibited by a very high ethanol concentration of 500 mg/dl. Production of reactive oxygen species by normal PMNs was assessed by either chemiluminescence (CL) for hypochlorous acid or ferricytochrome c reduction (FCR) for superoxide anions. For PMN stimulated by fMLP, ethanol inhibited CL but not FCR. For PMNs activated by phorbol myristate acetate, ethanol inhibited both CL and FCR. Ethanol did not alter baseline [Ca2+]i, as assessed by videomicroscopy using the Ca(2+)-sensing fluorescent dye Fura-2-AM, but did significantly potentiate the increase in peak [Ca2+]i levels that occurs in response to stimulation by fMLP. Calcium channel blockers attenuated ethanol's inhibition of CL. Thus, acute in vitro ethanol, at clinically relevant concentrations, can inhibit several critical aspects of PMN functions. But, in PMNs, unlike neural cells, these inhibitory effects do not seem to be mediated by decreases in Ca2+ influx or in [Ca2+]i. PMID- 8730219 TI - Neuronal degeneration in rat cerebrocortical and olfactory regions during subchronic "binge" intoxication with ethanol: possible explanation for olfactory deficits in alcoholics. AB - Severe, repetitive ("binge") ethanol intoxication in adult rats (intragastric delivery 3 times daily for 4 days in a modification of the Majchrowicz method) precipitates neuronal degeneration in selected cerebral cortical regions involved in memory and olfaction, confirming the results of Switzer and colleagues (Anat. Rec. 202: 186a, 1982). Neuronal damage was visualized with the de Olmos cupric silver technique for degenerating neurons and processes (argyrophilia), and was quantitated by total counts and densities of argyrophilic cells/fields. The specificity of the degeneration provides a neuropathological basis for the olfactory memory deficits in chronic alcoholics. In highly intoxicated rats, argyrophilia was most extensive among hippocampal dentate gyrus granule cells, pyramidal neurons in layer 3 of the entorhinal cortex, and olfactory nerve terminals in the olfactory bulb. Degenerating pyramidal neurons were also consistently seen in the insular cortex and olfactory cortical regions, such as the piriform and perirhinal cortices. There were few argyrophilic neurons in the CA regions of the hippocampus and none in the cerebellum--regions generally shown to have cell loss in long-term ethanol feeding models--but degenerating mossy fibers in the CA2 region were observed. Degeneration was maximal before the peak period of abstinence symptoms in this model, because argyrophilic densities were no greater 36 hr, compared with 8 hr after the last ethanol dose. High blood ethanol levels were required, because argyrophilia, absent from isocaloric controls, also was only evident in ethanol-intoxicated rats with mean blood ethanol levels for days 2 to 4 above 300 mg/dl; however, it increased substantially between 350 and 550 mg/dl. The resemblance of the argyrophilic distribution to the regional neuropathology that occurs in experimental seizures indicates that the ethanol-induced degeneration may have an excitotoxic basis. Progressive reductions in the seizure threshold (e.g., kindling phenomena that have been documented during binge ethanol intoxication) might be associated with excitotoxic hyperactivity during the repetitive nadirs between high blood and brain ethanol peaks. However, direct toxic actions of ethanol or its metabolites could also be involved. Overall, the model should be useful for studying mechanisms of ethanol-induced selective cortical and olfactory brain damage. PMID- 8730220 TI - Interleukin-6 tumor necrosis factor-alpha clearance and metabolism in vivo and by the isolated, perfused liver in the rat: effect of acute alcohol administration. AB - Plasma clearance and organ distribution of intravenously injected human recombinant [125I]interleukin (IL)-6 and [125I]tumor necrosis factor (TNF)-alpha were studied in male rats, 2 hr after intravenous alcohol (ethanol) administration (single dose, 2.2 g.kg-1 body weight). Also, the rate of uptake and degradation of the two cytokines by the isolated, perfused rat liver was studied in the absence or in the presence of ethanol (35 mM) in the perfusate. Acute ethanol administration significantly increased plasma clearance rate for both cytokines (36% and 72%, for IL-6 and TNF-alpha, respectively), decreased the t1/2 alpha (30% and 11%, for IL-6 and TNF-alpha, respectively), abolished the slow (beta)-phase component for TNF-alpha, and increased t1/2 beta for IL-6 (31%). Although alcohol did not affect organ distribution of TNF-alpha, it increased the IL-6 content in the liver, kidney, and blood. IL-6 uptake rate by the isolated, perfused rat liver was 2-fold higher than TNF-alpha uptake, whereas the rate of degradation was larger for TNF-alpha than for IL-6, despite the fact that both cytokines were presented to the liver at the same concentration (6 nM). Ethanol addition to the perfusate (35 mM, final concentration) significantly increased TNF-alpha uptake (24%), without affecting IL-6 uptake or the degradation rate of either cytokine. Also, the kinetics of degradation by the isolated, perfused rat liver was linear for TNF-alpha, but exponential for IL-6. Data presented in this study demonstrate that: (1) acute alcohol consumption can alter the kinetic behavior of IL-6 and TNF-alpha in the bloodstream, mainly by accelerating their clearance which, in turn, may counteract the outcome of cytokine secretion and delivery to the blood; and (2) short exposure of liver to ethanol levels commonly seen in humans after binge drinking may alter its capacity to take up cytokines. PMID- 8730221 TI - Role of protein synthesis on ethanol regulation of adenylyl cyclase activity in wild-type S49 murine lymphoma cells. AB - Adenylyl cyclase activity was determined in membranes from wild-type S49 murine lymphoma cells that had been exposed to ethanol for 4 hr. Mn-, NaF-, and forskolin-stimulated adenylyl cyclase activities of cells-pretreated with cycloheximide, puromycin, or serum deprivation-were significantly decreased by treatment with 50 mM of ethanol. As demonstrated for Mn-stimulated activity, the decrease was dose-dependent on ethanol and was temporal; a normal activity recovered after 16-24 hr treatment, even in the presence of cycloheximide and ethanol. Studies with a cell-free membrane system of S49 cells revealed a similar activity decrease after treatment of the membranes with ethanol. In contrast, cells treated with 50 mM of ethanol in a regular culture condition showed no decrease in adenylyl cyclase activity over 24 hr. These results indicate that ethanol regulation of adenylyl cyclase activity in S49 cells depends on reduced or impaired protein synthesis. PMID- 8730222 TI - Buspirone in the treatment of alcohol dependence: a placebo-controlled trial. AB - A double-blind controlled study of the effects of buspirone on alcohol dependence and associated symptoms in ambulatory alcoholics showed a marked improvement in both treatment and control groups. However, significant differences in favor of the medication were observed in several psychopathological measures, but not in measures of alcohol consumption. Contrary to a majority of prior controlled trials of buspirone in alcoholics, subjects were not selected on the basis of comorbid generalized anxiety; rather, the study tested the hypothesis of a direct effect on craving and consumption, independent from an anxiolytic effect. This hypothesis was not confirmed. PMID- 8730223 TI - Alcohol consumption, life context, and coping predict mortality among late-middle aged drinkers and former drinkers. AB - This study examined mortality risk for individuals in four alcohol consumption categories and identified life context and coping factors that independently predicted mortality among late-middle-aged drinkers and former drinkers (n = 1869). Compared with light drinkers, former drinkers (current abstainers) were at increased mortality risk; moderate drinkers were at decreased risk. Consistent with previous research on older samples, heavy drinkers were not at increased risk. Abstainers' increased risk was reduced in a model that controlled for life context and coping factors. Other independent predictors of mortality included reporting an illness stressor, stressor severity, less participation in activities with friends, greater use of resigned acceptance and alternative rewards coping, and less use of cognitive avoidance and emotional discharge coping. The findings support previous research on the alcohol-mortality relationship among older adults, and highlight the fact that abstainers' life stressors and avoidance coping responses may be more important predictors of their mortality than their abstention. PMID- 8730224 TI - Use of ICD-9-CM codes in the estimation of alcohol-involved injury: search for a surrogate II. AB - This study presents a follow-up analysis of 22,427 injury cases drawn from the California Regional Trauma Registry. Whereas the earlier analysis developed a surrogate for alcohol-involved injuries using E-codes as reflective of injury type, this analysis explores the possibility of using ICD-9-CM diagnosis codes that have a longer history and are available in more jurisdictions. Findings reported herein indicate that the original demographic patterns predicting testing and alcohol involvement patterns were maintained in ICD-9-CM diagnostic code-based models. Moreover, although variables representing demographic background, time of day, and day of week clearly were the most powerful model predictors, ICD-9-CM diagnostic codes reflecting physician assessment that the patient had a chronic alcohol problem did improve the fit of models, and thus provide additional information concerning testing and blood alcohol concentration patterns. In contrast, ICD-9-CM diagnostic codes reflecting the specific nature of the injury, although statistically significant, explained little additional variability in alcohol involvement. Nevertheless, the overall model did accurately classify approximately 75% of those in the sample for whom blood alcohol concentration status information was available, suggesting its appropriateness for surrogate development. PMID- 8730225 TI - Diabetic patients' alcohol use and quality of life: relationships with prescribed treatment compliance among older males. AB - One hundred fifty-four males attending an outpatient diabetes clinic were interviewed to determine the incidence of alcohol use among them and to identify mechanisms by which alcohol use adversely affects the control of diabetes. Specifically, we determined how diabetic patients' use of alcohol is related to compliance with their diabetes treatment regimen, and diabetic patients' expectations of benefits from drinking alcohol and actual alcohol use is related to their quality of life. Greater alcohol use was related to poorer prescribed insulin injection compliance (p < 0.01), and stronger expectations of immediate, positive consequences of drinking alcohol were related to several indices of lower quality of life (p < 0.005). These results imply interactive relationships among diabetics' alcohol use and expectancies, treatment compliance, and quality of life. PMID- 8730226 TI - Moderate alcohol consumption and disorders of human spermatogenesis. AB - Environmental factors are suspected to be responsible in part for the deterioration in semen quality observed worldwide during the recent few decades. Alcohol might be one factor, considering the frequent changes in testicular function associated with heavy drinking. The dose-dependent effects of alcohol on human spermatogenesis are, however, not well known. We analyzed spermatogenesis and testicular tissue morphology of 195 men, aged 35-69 years, with computer assisted microscopy in this autopsy study. The men were categorized into controls and four "consumption groups" according to the average daily alcohol consumption, which was determined on the basis of blind interviews with relatives and acquaintances. When the average daily alcohol consumption was 40 g or less, 59 (66%) of the 90 men showed normal spermatogenesis, whereas 31 (34%) had partial spermatogenic arrest (SA). Of the 31 men with average daily intake between 40 and 80 g, 17 (54%) showed normal spermatogenesis, 13 (42%) had partial or complete SA, and 1 man exhibited more severe testicular damage-"Sertoli cell only" (SCO) syndrome. Among men with daily intake between 80 and 160 g, only 13 of 35 men showed normal spermatogenesis (37%), 19 (54%) had partial or complete SA (odds ratio = 2.92), and 3 (9%) had the SCO syndrome (odds ratio = 16.85). The frequencies of spermatogenic disorders were similar in men drinking in excess of 160 g. Both SA and the SCO syndrome showed a clear dependence on daily dose; p < 0.0001 and p < 0.0004, respectively. We conclude that long-term average daily consumption of < 40 g of alcohol seems not to be associated with disorders of spermatogenesis. Consumption of moderate amounts of alcohol may affect semen quality more often than previously thought, whereas high alcohol consumption may even be associated with serious disorders of spermatogenesis. PMID- 8730227 TI - Alcohol in fatal and nonfatal injuries: a comparison of coroner and emergency room data from the same county. AB - Although alcohol is thought to be associated with severity of injury, relatively little data are available that compares alcohol's involvement in injury cases treated in the emergency room (ER) with coroner cases of injury fatality, both coming from the same county. Data from a probability sample of casualty patients 18 years and older treated at the University of Mississippi Medical Center ER in Jackson during a 6-month period (n = 275) are compared with data from coroner reports of all fatalities from unnatural causes among those 18 and older in the same county during a 1-year period surrounding the 6 months of data collection in the ER (n = 222). The two samples are compared on demographic characteristics, cause of injury, place of injury, and alcohol use before the event. A significantly larger proportion of the coroner sample was positive for alcohol (57%) and intoxicated (36%), compared with the proportion of those in the ER sample breathalyzed within 6 hr of injury, and reporting no drinking after the event who were positive (15%) and intoxicated (6%). Differences were most pronounced for motor vehicle accidents and fires. Violence-related injuries were more likely in the coroner sample (32%) than in the ER sample (16%), and they were more likely to involve alcohol at levels of intoxication. Those in the coroner sample were also more likely to be alcohol-positive for injuries occurring in all places except the home of another and the workplace. Using logistic regression, gender (male) and site (coroner) were predictive of a positive blood alcohol across all causes of injury combined. Gender (female), being alcohol-positive and site (coroner) were significantly predictive of motor vehicle accidents. Alcohol was not found to be a significant predictor for falls, other accidents, or injuries resulting from violence. Data suggest that alcohol's association with severity of injury varies by cause of injury. PMID- 8730228 TI - Effect of diazepam on plasma gamma-aminobutyric acid in sons of alcoholic fathers. AB - A subgroup of abstinent alcoholics, display low levels of plasma gamma aminobutyric acid (GABA). Two previous studies of plasma GABA in sons of alcoholic fathers (SOAs) have yielded conflicting results. The aim of the current study was to measure plasma GABA both at baseline and after challenge with diazepam, a GABAA receptor agonist, in a group of SOAs already shown to display decreased eye movement, memory, and sedative effects of diazepam. Twenty-seven SOAs and 23 male control subjects received four logarithmically increasing doses of diazepam or placebo in randomized order on 2 days at least 1 week apart. Plasma GABA was measured at baseline and after the last dose. There were no significant differences between SOAs and controls in baseline plasma GABA levels. In the whole sample, there were significant correlations between baseline plasma GABA and both high novelty-seeking and low-harm avoidance scores on the Tridimensional Personality Questionnaire. Both SOAs and controls displayed decreases in plasma GABA over time on both testing days, but there was no effect of diazepam on plasma GABA and no significant difference between groups in plasma GABA response to diazepam. These results suggest that neither low plasma GABA at baseline nor altered plasma GABA response to diazepam is associated with increased genetic risk for alcoholism. PMID- 8730229 TI - Relationship between alcohol withdrawal seizures and temporal lobe white matter volume deficits. AB - A previous magnetic resonance imaging study from our laboratory reported significant temporal lobe volume deficits in cortical gray matter, white matter, and anterior hippocampus in chronic alcoholic men relative to controls. In the present study, we reexamined these data and asked whether withdrawal seizure history was predictive of either the hippocampal or the extrahippocampal volume deficits. A review of the medical charts indicated that 11 alcoholics had experienced one or more alcohol-related seizures and 35 were seizure-free; no patient had a seizure disorder unrelated to alcohol. The two alcoholic groups did not differ significantly in age, education, alcohol consumption variables, premorbid intelligence, Memory Quotient, Trail Making, or detection of hidden figures. Although each alcoholic group showed significant bilateral volume deficits of the anterior hippocampus and frontal-parietal and temporal gray matter, relative to controls, the seizure group had significantly smaller temporal lobe white matter volumes than either the control or the seizure-free groups; the latter two groups did not differ from each other. Both alcoholic groups, however, had white matter volume deficits in the frontal-parietal region. Thus, the seizure group accounted for the white matter volume deficits in the temporal lobe previously reported in the full sample of alcoholics. It seems, then, that reduced white matter volume in the temporal lobes may be either a risk factor for or sequela of alcohol withdrawal seizures. PMID- 8730230 TI - Increased vulnerability to alcohol-related birth defects in the offspring of mothers over 30. AB - The risk of fetal alcohol syndrome (FAS) is known to increase with increased maternal age and parity. This study investigated the hypothesis that the deficits in growth and intellectual function seen in non-FAS infants exposed to alcohol at moderate-to-heavy levels are also found disproportionately in the offspring of older mothers. Mothers of 480 African-American, inner-city infants were interviewed at each prenatal clinic visit regarding their use of alcohol during pregnancy. Infants were assessed for physical growth and cognitive development repeatedly through age 13 months. In analyses run separately for the infants of younger and older mothers, alcohol-related deficits were seen most strongly in the offspring of women over 30 years of age. This pattern was not caused by lower levels of drinking by the younger mothers. Age-related increases in maternal body fat-to-water ratio and a faster rate of alcohol metabolism in chronic drinking women may account for the greater vulnerability of the offspring of the older mothers. These data suggest that physiological changes associated with aging and/or chronic drinking may play an important role in the alcohol-related birth defects seen in infants exposed at moderate-to-heavy levels. PMID- 8730232 TI - Toward an alcohol treatment model: a comparison of treated and untreated respondents with DSM-IV alcohol use disorders in the general population. AB - The purpose of this study was to compare characteristics of person with alcohol use disorders who sought alcohol treatment with those who did not using data from a nationally representative sample of the United States. Applying an organizing framework from the larger literature on service utilization, a logistic regression analysis was conducted to examine the interaction among factors influencing treatment. The results identified unemployment status and lower educational level as barriers to alcohol treatment, but the impact of these factors differed depending on whether the respondent had previous experience with alcohol treatment. The major findings of this study are discussed in terms of consumer satisfaction, minimizing barriers to alcohol treatment services, and the need to examine individual determinants of alcohol treatment within the larger context of organizational and sociopolitical factors. PMID- 8730231 TI - Alcohol use and T-lymphocyte subsets among injection drug users with HIV-1 infection: a prospective analysis. AB - OBJECTIVES: Alcohol use is known to alter immune function and has immunosuppressive effects that may modify T-lymphocyte subpopulations. However, there is no clear evidence regarding the relationship of alcohol use with the progression of immunodeficiency in human immunodeficiency virus-type 1 (HIV-1) seropositive individuals, particularly injection drug users (IDUs). METHODS: Using prospective data from a cohort of IDUs in a study of the natural history of HIV infection, we examined the relationship of alcohol use and changes in T lymphocyte subsets. Among the 2921 IDUs followed semiannually in outpatient clinics, 188 were documented HIV-1 seroconverters with known time of seroconversion. At each visit, all study participants were interviewed, underwent physical examinations, and had blood drawn for laboratory studies. Alcohol use was measured by reported frequency and quantity of alcoholic beverages. Longitudinal analyses included data for up to 5 years postseroconversion. To formally test the association of alcohol use with change in levels of CD4+ and CD8+ cells subsequent to HIV seroconversion, regression models incorporating autocorrelation structure were applied. RESULTS: Alcohol use was not appreciably related to age, gender, marital status, income, education, or the duration of intravenous drug use. CD4% decreased for all IDUs within the first 5 years after seroconversion, with no significant differences between alcohol categories. CD8% increased for all IDUs, with no significant differences by alcohol category within the first 2 years after seroconversion. However, between 2 to 5 years postseroconversion, there was a statistically significant increase among the heaviest drinkers: CD8% increased 6.9%/year [95% confidence interval (CI): 4.7, 8.0] for the IDUs who reported > 21 drinks/week, 2.4%/year (95% CI: 0.8, 4.0) for IDUs who drank 21 drinks/week or less, and 0.4% (95% CI: -2.1, 2.9) for abstainers. Similar results were obtained for CD4 and CD8 absolute counts. CONCLUSIONS: In this study population of IDUs, CD8% (but not CD4%) is associated with alcohol consumption early after HIV seroconversion. This is the first prospective study to date to assess the relationship of alcohol use with HIV progression from the time of HIV seroconversion among a cohort of IDUs. If confirmed in future investigations, the findings may have significant implications for prevention and early intervention programs aimed at inhibiting disease progression among HIV-positive IDUs. PMID- 8730233 TI - Alcohol and substance use among Japanese high school students. AB - In the first large-scale survey on the use of alcohol and tobacco and the inhalation of paint thinner by Japanese high school students, a higher percentage of respondents reported past or regular use of these substances than in a 1978 survey. The survey was conducted in 44 high schools. The 14,438 respondents (ages 15-18 years) represented 9 of Japan's 47 geographically distinct urban and rural prefectures. Most (76%) reported that they had consumed alcohol at some time, and 50% reported drinking on a regular basis. Alcohol consumption, drinking partners, and motivation for drinking were compared by student gender, geographic region, and years of education. Fewer students reported smoking cigarettes: 32% had smoked at some time, and 14% smoked regularly. Inhalation of paint thinner was rare: 3% had purposely inhaled it, and 1% had abused it. A significant percentage of the alcohol drinkers also used tobacco and/or thinner, a tendency that was the same despite differences in gender, grade in high school, and urban or rural geographic region. A tendency toward polysubstance use was evident among these high school students. PMID- 8730234 TI - DSM-IV alcohol dependence in a treatment sample of white, black, and Mexican American men. AB - This study examines the presentation, correlates, and factor structure of DSM-IV alcohol dependence among 256 White, 263 Black, and 212 Mexican-American men admitted consecutively to five alcohol treatment programs in San Jose, CA. Interviews of approximately 1 hr were conducted in the programs' facilities by trained interviewers using a standardized questionnaire. The response rate was 87%. Results show that the proportion of Black respondents who are alcohol dependent according to the DSM-IV criteria is lower (63%) than the proportion of Whites (86%) and Mexican Americans (76%). However, the proportion of respondents reporting each criterion of dependence was similar across groups. The most powerful predictor of the number of dependence indicators reported by respondents was level of alcohol consumption, independent of ethnicity. A unidimensional model of dependence combining all seven indicators of DSM-IV alcohol dependence fit well across men in all three ethnic groups. These results indicate that both the presentation and factorial structure of DSM-IV alcohol dependence were uniform across White, Black and Mexican-American men in treatment for alcohol problems. PMID- 8730235 TI - Conference summary: Consensus Conference on Alcohol Dependence and the Role of Pharmacotherapy in its Treatment. AB - Alcoholism, a major public health problem throughout the world, causes enormous damage to health and quality of life and undermines the well-being of families and society. It is associated with liver disease, cancer, cardiovascular problems, accidental deaths, suicides, and homicides. Because of the internationally recognized severity of this disease, this consensus conference was convened to bring together leading international experts to share ideas with regard to current research in the areas of alcohol dependency and treatment, and to discuss the future directions for the pharmacotherapy of alcoholism. The work presented at this conference points to a substantial evolution in the ability of researchers to understand the mechanism by which alcohol acts on the brain and reinforces the concept that alcoholism is a disease of brain chemistry, with a hereditary predisposition that is triggered by psychological and/or environmental factors that are thus far unidentified. The consensus statements proposed by the participants at the close of the conference highlight the definition, etiology, and prevention of alcoholism, as well as current and potential treatments and recommendations that can be used to guide future pharmacologic research. PMID- 8730236 TI - Voice of the Victims. AB - Over the past 10 years, I have been privileged to conduct educational forums for audiences containing many recovering alcoholics or otherwise chemically dependent persons. In these forums about the addictive diseases and their treatment and research possibilities, significant interaction with the audience members occurs. During these interactions, certain anecdotal phenomena seem to predominate. The repetitive nature of these reports suggests in the need for systematic investigation. As with editorial comments in major medical journals, observed phenomena and unanswered questions from the victims can be valuable in the generation of testable hypotheses. Perhaps the ideas presented herein will be useful in the development of future research on alcohol abuse and alcohol dependence. PMID- 8730237 TI - Ethanol alters spatial processing of hippocampal place cells: a mechanism for impaired navigation when intoxicated. AB - This study describes a new mechanism by which ethanol alters brain function and may impair performance on tasks requiring spatial navigation. Recording electrophysiological activity from single neurons in the awake, freely behaving animal, the present study shows that ethanol impairs the ability of place cells in the hippocampus to process spatial information. The impairment by ethanol in spatial processing of place cells was remarkably similar to the impairment produced by lesions of afferents to the hippocampus, except that the effect of ethanol was reversible. Since lesions to hippocampal afferents that alter spatial processing of place cells concomitantly impair spatial navigation, the present results suggest that ethanol similarly impairs spatial navigation by altering spatial processing of place cells. The present results have implications for the observation that ethanol impairs performance on navigational tasks that require spatial processing, such as automobile driving. PMID- 8730238 TI - 8-Bromo-cAMP mimics beta-adrenergic sensitization of GABA responses to ethanol in cerebellar Purkinje neurons in vivo. AB - Previous studies in our laboratory indicated that electrophysiological responses of cerebellar Purkinje neurons to GABA were not routinely potentiated by ethanol (EtOH), and the potentiation was not large when it occurred. In the presence of beta-adrenergic agonists, such as isoproterenol, however, GABA inhibitions became sensitive to potentiation by EtOH in nearly every Purkinje neuron tested. beta adrenergic receptor activation alone also modulates (potentiates) GABA responses on Purkinje neurons, and this has been reported to be mediated by a cAMP second messenger system. Herein, we report that the membrane-permeable cAMP analog, 8 bromoadenosine-3',5'-cyclic monophosphate (8-Br-cAMP), but not the membrane impermeable cAMP, can also modulate GABA responses and that EtOH potentiates this facilitatory action of 8-Br-cAMP. These effects are not likely caused by adenosine receptor mechanisms, because this 8-bromoadenosine mediated modulation and sensitization was observed in the presence of systemic theophylline. These data suggest that the beta-adrenergic modulation and sensitization to EtOH of cerebellar Purkinje neuron GABA responses occur via a cAMP second messenger mechanism. PMID- 8730239 TI - Review article: quantitative leucocyte scanning in the assessment of inflammatory bowel disease activity and its response to therapy. AB - Inflammatory bowel disease (IBD) symptoms may be due not only to the effects of bowel inflammation, but also can result from many non-inflammatory consequences of the disease. Clinical scores and health questionnaire ratings produce a global assessment which is useful in the overall evaluation of the impact of the illness on the patient, and the effect of treatment. However many studies have failed to recognize the need to separately assess changes in the degree of bowel inflammation, in addition to a global clinical response. Radiolabelled white cell scanning using 111-indium has been shown to provide an accurate, quantitative and non-invasive method for assessing the degree of bowel inflammation in IBD and its response to therapy, using faecal collection or total body counts of radioactivity. More recently 99mTc hexamethyl propylene amine oxime (HMPAO) labelled white cell scanning has been introduced, which has advantages over the 111-indium method, including reduced radiation dosimetry and enhanced image quality. 99mTc-HMPAO scanning using three-dimensional white cell scanning (single photon emission computerized tomography; SPECT) allows visualization of the entire bowel separate from overlying structures. 99mTc-HMPAO white cell scanning with SPECT has now been computerized and automated to permit measurement of segmental and total bowel radiolabelled white cell infiltrate. This method could potentially provide the gold standard for objective assessment of the response of disease activity in Crohn's disease and ulcerative colitis, and merits application in clinical trials of novel therapies for IBD. PMID- 8730240 TI - Gastric mucosal injury and adaptation to oral and rectal administration of naproxen. AB - INTRODUCTION: Oral nonsteroidal anti-inflammatory drugs (NSAIDs) cause acute gastric mucosal injury but the relative importance of systemic and topical effect of NSAIDs to overall gastric damage remains uncertain. METHODS: Twenty-four healthy volunteers were allocated either oral or rectal naproxen 500 mg b.d. and gastroscoped before and during days 1, 7 and 28 of dosing. Macroscopic gastric damage was assessed using a modified Lanza score, mucosal blood flow recorded using laser Doppler flowmetry and prostaglandin E2 (PGE2) measured in antral mucosal biopsies. RESULTS: Maximal gastric damage occurred during the first 24 h in the oral naproxen group and was associated with a fall in antral mucosal blood flow (mean +/- S.E.M.) from 58.2 +/- 3.3 to 46.6 +/- 4.1 arbitrary units (a.u.) (P < 0.05). With continued administration of oral naproxen, gastric damage resolved and antral mucosal blood flow returned to baseline (54.2 +/- 3.7 a.u.). No macroscopic damage or significant changes in mucosal blood flow were observed during rectal administration. There was no significant difference between mucosal PGE2 concentrations in those receiving oral or rectal naproxen, falling from an initial level of 335 +/- 29 to 155 +/- 49 pg/mg at day 1 (P = 0.06) in those receiving oral naproxen and from 235 +/- 55 to 107 +/- 31 pg/mg at day 1 (P = 0.1) in those receiving rectal naproxen, and remaining suppressed throughout the study in both groups. CONCLUSIONS: These observations suggest that acute mucosal damage and changes in mucosal blood flow are caused by the topical rather than systemic actions of naproxen. PMID- 8730241 TI - Eradication of Helicobacter pylori in general practice. AB - OBJECTIVES: To determine the current management of patients with Helicobacter pylori infection in general practice with regard to the selection of patients, testing for the presence of the organism, the choice of eradication therapy and the occurrence of symptoms during follow-up. DESIGN: Cross-sectional survey involving the retrospective collection of data from patient medical records. SETTING: Five general practices in the Fife region of Scotland during June and July 1995. MAIN OUTCOME MEASURES: The proportion of patients appropriately selected for eradication therapy and the success of treatment as assessed by remission of symptoms during follow-up. RESULTS: Of 154 patients studied, 80% received eradication therapy for peptic ulcer disease, the remainder for non ulcer dyspepsia or gastrooesophageal reflux. Fifty-six different regimens were used, the most common combination of drugs being omeprazole plus amoxycillin. H. pylori status was known in only one-third of patients before treatment and in only 15% after treatment. More than half of patients complained of recurrent symptoms of dyspepsia during follow-up after eradication therapy and 47% required further treatment. In terms of the selection of patients and testing for H. pylori, the overall management using eradication therapy was acceptable in less than half of patients. In particular, the management of patients with ulcer disease associated with either ulcerogenic drugs or previous complications failed to provide the essential protection against the high risk of future haemorrhage or perforation. CONCLUSIONS: The findings of this study indicate an urgent need for improved education if therapy for the eradication of H. pylori is to be used appropriately and safely. PMID- 8730242 TI - Repeat prescribing of ulcer healing drugs in general practice--prevalence and underlying diagnosis. AB - BACKGROUND: The long-term use of ulcer healing drugs in the management of dyspepsia is controversial. We have investigated repeat prescribing of these drugs in a general practice population. AIMS: To identify the number of patients authorized to receive repeat prescriptions for ulcer healing drugs, and to review the investigation status and diagnosis in these patients. SUBJECTS: A total of 15,495 patients registered with eight general practitioners in seven general practices in Dundee, UK. METHODS: Case ascertainment by review of practice repeat prescribing registers. Data regarding investigation and diagnosis obtained by retrospective review of general practice case records. RESULTS: Six hundred and seventy-nine (4.4% of the total population) were authorized to receive repeat prescriptions for ulcer healing drugs. Six hundred and fifty-one (4.2%) were authorized to receive repeat prescriptions for H2-antagonists. Ranitidine was prescribed in 583 (86% of patients receiving ulcer healing drugs). Endoscopy had been performed in 426 (63%) and barium meal alone in 113 (17%); 140 (21%) had not been investigated. A diagnosis of peptic ulcer disease or oesophagitis was established in 382 (56%). However, 157 investigated patients (23% of all patients on ulcer healing drugs) did not have a peptic diagnosis. CONCLUSIONS: The prevalence of repeat prescribing of ulcer healing drugs in the general practice population studied was 4.4%, but 44% of these patients did not have a confirmed diagnosis of acid peptic disease. PMID- 8730243 TI - Gastrointestinal tolerability of lornoxicam compared to that of naproxen in healthy male volunteers. AB - BACKGROUND AND AIM: Gastrointestinal effects are common adverse effects associated with nonsteroidal anti-inflammatory drugs (NSAIDs). Lornoxicam is a new nonsteroidal anti-inflammatory drug and its gastroduodenal tolerability was compared to that of naproxen in a randomized, double-blind, crossover study. METHODS: Eighteen healthy male volunteers received lornoxicam 8 mg b.d. or naproxen 500 mg b.d. administered orally over two 7-day dosing periods. Upper endoscopy was performed by two independent investigators at the beginning and end of each dosing regimen. RESULTS: Lornoxicam 8 mg b.d. caused significantly less mucosal injury than naproxen 500 mg b.d. in the stomach/duodenal bulb, as well as in the mid/distal duodenum. CONCLUSION: These findings may have favourable implications for lornoxicam in the clinical setting, if this dose provides optimal control of arthritic pain. PMID- 8730244 TI - Sulphasalazine treatment and the colorectal mucosa-associated flora in ulcerative colitis. AB - AIM: To study the influence of sulphasalazine treatment on the mucosa-associated bacterial flora of rectal biopsy tissue specimens in patients with ulcerative colitis. PATIENTS: Twenty-four patients had newly diagnosed active ulcerative colitis; 20 patients had acute relapse of ulcerative colitis (10 not taking maintenance sulphasalazine); (40 patients had quiescent ulcerative colitis; 21 not taking maintenance sulphasalazine). The influence of 3 weeks of sulphasalazine treatment on the mucosa-associated flora was studied in the patients presenting with active disease. RESULTS: Comparison of patients according to sulphasalazine usage revealed few differences in the mucosal flora. In patients with quiescent ulcerative colitis, Escherichia coli was found at lower counts in patients taking maintenance sulphasalazine; however, this effect was not evident in patients with active disease. Inconsistent changes in other facultatives were seen between the two active disease groups, particularly for a miscellaneous group of unidentified Gram-positive rods. Three patients, all receiving sulphasalazine, were colonized with Clostridium difficile, but this did not appear to influence their disease. CONCLUSION: Sulphasalazine treatment in ulcerative colitis causes only minor disturbance to the populations of bacteria colonizing the colorectal mucosa. PMID- 8730245 TI - Treatment of Helicobacter pylori infection with low or high dose omeprazole combined with amoxycillin and the effect of early retreatment. AB - BACKGROUND: Cure rates of H. pylori infection, using dual therapy with omeprazole and amoxycillin, vary considerably and the efficacy of retreatment with this regimen in the case of initial failure is controversial. Therefore, we conducted a large prospective double-blind randomized trial, studying the efficacy of low vs. high dose omeprazole in dual therapy and of early retreatment with the same regimens. METHODS: One hundred and sixty-eight consecutive H. pylori-positive patients, suffering from either peptic ulcer disease or functional dyspepsia, were enrolled. Group I (n = 84) received omeprazole 20 mg b.d. plus amoxycillin 750 mg t.d.s., for 2 weeks. Group II (n = 84) received omeprazole 40 mg t.d.s. plus amoxicillin 750 mg t.d.s., for 2 weeks. RESULTS: The H. pylori eradication rate was 60.2% in group I and 64.3% in group II (P = 0.59). Cure of H. pylori infection was significantly better in patients with peptic ulcer disease, compared to non-ulcer dyspeptics (P = 0.016). Retreatment, given in 54 patients, was successful in 21.4% patients in group I and in 28% patients in group II (P = 0.58). CONCLUSIONS: High dose of omeprazole has no advantage compared to low dose in terms of eradication efficacy. Early retreatment with the same regimen offers limited improvement in cure rate. Presence of peptic ulcer disease influences cure rates significantly. PMID- 8730246 TI - Mast cells do not contribute to nonsteroidal anti-inflammatory drug-induced gastric mucosal injury in rodents. AB - BACKGROUND: By releasing pro-ulcerogenic mediators, mast cells may contribute to the mucosal injury associated with the use of nonsteroidal anti-inflammatory drugs (NSAIDs). METHODS: To study this, rat and mouse models of NSAID-induced gastric damage were used in which administration of indomethacin causes haemorrhagic injury in the corpus region of the stomach, and the "re-feeding" model in which penetrating antral ulcers are induced in the rat by naproxen. Mast cell degranulation was determined histologically and by measurement of tissue and serum levels of rat mast cell protease-II, a mediator specific to mucosal mast cells. The effects of either increasing or decreasing the number of gastric mucosal mast cells on the susceptibility of the stomach to injury induced by indomethacin were also studied. RESULTS: Gastric injury induced by indomethacin was not accompanied by significant mast cell degranulation. Moreover, neither increasing nor decreasing the number of gastric mucosal mast cells had a significant effect on the susceptibility of the gastric mucosa to damage induced by indomethacin. In the re-feeding model, prior depletion of gastric mucosal mast cells did not significantly affect the severity of antral ulceration induced by naproxen, nor the ability of prostaglandin E2 to prevent this damage. Finally, indomethacin-induced damage was similar in severity in mice with a genetic defect resulting in the complete absence of mast cells as it was in normal, congenic littermates. CONCLUSION: Mast cells do not play a significant role in the development of gastric injury induced by acute NSAID administration in the rat or mouse. PMID- 8730247 TI - Gastric mucosal infiltration by Helicobacter pylori favours bacterial survival after treatment. AB - BACKGROUND: Although a number of patient and bacterial factors have been identified as predictors of treatment failure in Helicobacter pylori-associated gastritis, the causes of lack of response to treatment have not been fully elucidated. We hypothesized that bacterial infiltration of the gastric mucosa might be one of the factors responsible for treatment failure in patients harbouring the bacterium. METHODS: We studied 182 patients with gastritis who underwent anti-H. pylori treatment with different drugs. Gastric biopsies obtained at endoscopy, were examined for electron microscopic features of infiltration and damage. Patients were assigned to different treatment groups, and endoscopy, evaluation of H. pylori status and electron microscopy were repeated at least 4 weeks after the end of treatment. RESULTS: The overall H. pylori eradication rate was 65%. Eradication was achieved more frequently in patients without electron microscopic features of infiltration (85%), than in those patients with the bacteria deeply embedded into the gastric mucosa (45%; P < 0.0001). No treatment appeared to be clearly superior for patients with the highest degree of mucosal infiltration. CONCLUSIONS: Bacterial mucosal infiltration may facilitate the survival of H. pylori during antibacterial treatment; moreover, electron microscopy may be helpful to identify patients potentially unresponsive to anti-H. pylori treatment. PMID- 8730248 TI - Twenty-four-hour intragastric acidity following early evening or bedtime administration of roxatidine in duodenal ulcer patients. AB - AIM: To assess the usefulness of early evening administration of roxatidine 150 mg as an alternative to the traditional bedtime regimen. METHODS: Twenty-four patients with healed duodenal ulcer were dosed according to a balanced incomplete block design, with two of the following regimens: placebo, roxatidine 150 mg at 07.30 h (early evening) or roxatidine 150 mg at 22.00 h (bedtime). Twenty-four hour intragastric pH-metry was started at 18.00 h on the first day of dosing. Median pH was determined for the 24-h period, and for the following time intervals: 20.00-00.00 h, 00.00-08.00 h and 08.00-18.00 h. Percentage time in the 24-h period with pH greater than 4.0 was also calculated. RESULTS: The two roxatidine regimens proved significantly superior to the placebo, decreasing 24-h acidity for all the time intervals, except the 20.00-00.00 h period, when mean intragastric pH with the early evening regimen (4.5 +/- 1.1) proved significantly higher than after placebo (2.2 +/- 1.0) or when roxatidine was taken at bedtime (2.4 +/- 1.1). CONCLUSION: Early evening administration of roxatidine may afford satisfactory control of 24-h acidity, offering a useful alternative to conventional bedtime administration. PMID- 8730249 TI - Triple therapy with sucralfate, amoxycillin and metronidazole for healing duodenal ulcer and eradicating Helicobacter pylori infection. AB - METHODS: Seventy-seven chronic duodenal ulcer patients (50 male) were entered into this study. Treatment was started with sucralfate suspension (2 g b.d.) for 8 weeks. After 2 weeks the patients also received 750 mg amoxycillin t.d.s. plus 500 mg metronidazole t.d.s. for 12 days. Endoscopy with six antral biopsies (urease test, Gram staining, culture and histology) was performed before commencement of sucralfate therapy, 4 weeks after the end of antibiotic therapy, and during the follow-up examinations at 6 and 12 months. RESULTS: Seven patients were excluded prematurely from the study. Helicobacter pylori in five patients had primary resistance to metronidazole and these patients were also excluded. The ulcer healing rate 4 weeks after the end of antibiotic therapy was 92% and the H. pylori eradication rate was 82% (all per protocol). In all patients who were still H. pylori-positive, the bacterium became resistant to metronidazole and histologically the inflammatory state of the mucosa was the same as before treatment. All H. pylori-eradicated patients (n = 53) were re-examined after 6 and 12 months; no ulcer recurrence was observed and each time only one reinfection was found. CONCLUSIONS: In an open study, sucralfate with amoxycillin and metronidazole appeared to act together to eradicate H. pylori infection and to speed duodenal ulcer healing. PMID- 8730250 TI - Viral nucleotide changes in asymptomatic hepatitis B carriers undergoing interferon treatment. AB - BACKGROUND: Interferon might induce mutation in regions of hepatitis B virus DNA that encode for immunologic target of cytotoxic T lymphocytes. AIM: To investigate the short-term effect of steroid priming and interferon therapy on hepatitis B virus, we followed the nucleotide changes in the precore and core region of hepatitis B virus DNA in seven healthy asymptomatic carriers who underwent steroid priming followed by recombinant alpha interferon for 3 months. METHODS: Hepatitis B virus DNA from serial sera of the patients were polymerase chain reaction-amplified, and the precore and core region directly sequenced and analysed. RESULTS: Analysis revealed no serial changes in the hepatitis B virus nucleotide sequence in any of the patients. CONCLUSIONS: Steroid priming and short-term treatment with interferon in healthy asymptomatic patients does not select for hepatitis B virus with mutations in the precore and core region. PMID- 8730251 TI - High effectiveness and safety of one-week antibiotic regimen in Helicobacter pylori eradication. AB - BACKGROUND: Helicobacter pylori is strongly associated with peptic ulcer: H. pylori eradication markedly decreases the recurrence rate of duodenal and gastric ulcer, but the optimum length of antibiotic therapy in the eradication of H. pylori is still unclear. AIM: To verify the effectiveness and side-effect profile of an eradicating regimen consisting of omeprazole 20 mg daily for 4 weeks and, during the first week, combination antimicrobial treatment with tinidazole 500 mg b.d. plus clarithromycin 250 mg b.d. in patients with active duodenal and gastric ulcer. METHODS: One hundred and ninety-six duodenal ulcer patients and 27 gastric ulcer patients with H. pylori infection were admitted into an open prospective study. Compliance was assessed by an accurate interview. RESULTS: Overall, H. pylori was successfully eradicated in 201 of 223 patients (intention-to-treat 90.1%; 95% CI = 85-94%): 176 of 196 duodenal ulcer patients became H. pylori negative (89.8%; CI = 85-94%) as well as 25 of 27 gastric ulcer patients (92.6%; CI = 76-99%). Compliance was excellent in 221 of 223 (99.1%) patients evaluated as having taken all the medication as prescribed. Sixteen patients (7.2%) developed mild side effects during treatment. CONCLUSION: This combination treatment had excellent results with almost absolute compliance and a very low rate of minor side effects. PMID- 8730252 TI - One-week triple therapy with omeprazole, amoxycillin and either clarithromycin or metronidazole for cure of Helicobacter pylori infection. AB - AIM: The present study was designed to evaluate the efficacy and tolerability of 1-week triple therapy regimens for Helicobacter pylori. METHODS: In two consecutive series, 120 patients with proven H. pylori infection and peptic ulcer disease or functional dyspepsia were treated with either omeprazole 20 mg b.d., amoxycillin 1 g b.d. and clarithromycin 250 mg b.d. (OAC; n = 60) or with omeprazole 20 mg b.d., amoxycillin 1 g b.d. and metronidazole 400 mg b.d. over 1 week (OAM; n = 60). H. pylori infection was assessed by rapid urease test, culture and histology before and 4 weeks after cessation of the eradication therapy. RESULTS: H. pylori eradication succeeded in 53 out of 60 patients by omeprazole-amoxycillin-clarithromycin (OAC) (88%; 95% CI 77-95%) and in 47 out of 60 patients by omeprazole-amoxycillin-metronidazole (OAM) (78%; 95% CI 66-88%) (P = 0.22). Nine patients of each group available for follow-up reported adverse events (15.0 and 15.5%, respectively) without necessity of discontinuation of the study medications. Serious adverse events were not observed. CONCLUSIONS: Simple and convenient 1-week triple therapies consisting of omeprazole, amoxycillin and either clarithromycin or metronidazole are sufficiently effective in eradicating H. pylori infection. PMID- 8730253 TI - Comparison of two lansoprazole-antibiotic combinations (amoxycillin or classical triple therapy) for treatment of H. pylori infection in duodenal ulcer patients. AB - AIM: To compare the eradicating capacity of two different antibiotic-lansoprazole combinations (amoxycillin vs. standard triple therapy) with that of lansoprazole alone in Helicobacter pylori-positive duodenal ulcer patients. METHODS: Ninety six out-patients with H. pylori-positive duodenal ulcer were randomly assigned to receive one of the following three antiulcer regimens: (1) lansoprazole 30 mg b.d. for 4 weeks plus amoxycillin 1 g t.d.s. during the last 2 weeks; or (2) lansoprazole 30 mg once daily for 4 weeks plus classical triple therapy (tripotassium dicitratobismuthate 240 mg b.d., amoxycillin 1 g t.d.s. and tinidazole 500 mg b.d.) for the last 2 weeks; or (3) lansoprazole 30 mg once daily for 4 weeks. Endoscopy was repeated at the end of treatment and 1 month later. A rapid urease test and histology were used to determine H. pylori status. RESULTS: Duodenal ulcer healing rates at 4 weeks were 96% after both lansoprazole with amoxycillin, and lansoprazole with triple therapy, and 97% after lansoprazole alone. Eradication of H. pylori was significantly better with lansoprazole with triple therapy than with either lansoprazole with amoxycillin or lansoprazole alone (90% vs. 55% vs. 3%, respectively). CONCLUSION: Classical triple therapy combined with lansoprazole is significantly more effective than the lansoprazole with amoxycillin combination for the eradication of H. pylori in duodenal ulcer patients pre-treated with lansoprazole. PMID- 8730254 TI - Do non-steroidal anti-inflammatory drugs have an effect on gastric cell turnover? PMID- 8730255 TI - Adherence of Helicobacter pylori. AB - Helicobacter pylori infects only gastric type epithelium, to which it adheres closely and forms attaching-effacing lesions. Similar lesions are also seen in ferrets infected with Helicobacter mustelae, the only other host in which peptic ulcer occurs during the course of infection. These observations imply a specific interaction between bacterial adhesin(s) and host receptor(s). They account for the adherence of H. pylori to the gastric epithelium and suggest that adherence is an important virulence factor. Numerous experimental methods, each with their own advantages or disadvantages, are available to study the interaction and characterize the nature of bacterial adhesins and their receptors. Many studies have shown that H. pylori is able to agglutinate red blood cells. Amongst several bacterial lectins, at least one adhesin has a sialic acid receptor specificity. However, the gene encoding this putative adhesin is larger than originally thought, whilst the function and location of its product is unclear. Adherence of H. pylori to a wide variety of cell lines (gastric and non-gastric) has also been shown, and is necessary for the induction of pro-inflammatory responses. However, there is little evidence to support an association between haemagglutination and extent or pattern of adherence to gastric epithelium or gastric-derived cell lines. PMID- 8730256 TI - Biochemical aspects of Helicobacter pylori colonization of the human gastric mucosa. AB - Unlike Helicobacter felis and other Helicobacter species of animal origin, Helicobacter pylori colonizes the lower gastric mucin layer of the stomach and adheres to human gastric epithelial cells. It is still an open question if H. pylori can interact with specific glycoconjugates in the gastric mucin layer. It is possible that colonization of the oral cavity is a first step of a complex infectious process. Most likely resting or slow growing cells of H. pylori interact with Lewis blood group substances in the gastric mucin layer and on the epithelium. This initial colonization is probably followed by binding to specific cell surface glycoconjugates (glycoproteins and glycolipids such as GM3) and specific sialylated or highly sulphated molecules such as cell surface sulphatides and heparan sulphate. H. pylori may also bind to specific phospholipid molecules such as phosphatidylethanolamine on the gastric cells. The adhesion process of certain strains can stimulate 'close' cell adhesion including pedestal formation similar to the phenomenon typical for a special class of enterovirulent Escherichia coli called attaching effacing E. coli. After gastric cell destruction by ammonia and H. pylori toxins (such as the vacuolating toxin) H. pylori may colonize the extracellular matrix (ECM). This phenomenon seems to include binding of cell surface sialic acid specific haemagglutinin to one ECM component, i.e. laminin. It is also likely that H. pylori may use similar events to penetrate intercellular junctions of gastric epithelial cells. These adhesion penetration phenomena also involve coating of the microbe with host proteins to escape the host immune system and initiate a chronic lifelong infection process. PMID- 8730257 TI - Gastric mucosal inflammatory responses to Helicobacter pylori. AB - Within the gastroduodenal mucosa Helicobacter pylori infection stimulates local production of a range of proinflammatory and immunoregulatory cytokines, neutrophil infiltration, specific T- and B-cell responses and the development of gastric lymphoid follicles. Following bacterial eradication this mucosal inflammatory response resolves. Infiltrating neutrophils are likely to be one of the major mediators of mucosal damage. Neutrophil activation, including reactive oxygen metabolite production and the release of myeloperoxidase, will be induced directly by bacterial factors and indirectly through products of complement activation, bioactive lipids and host-derived cytokines. Interleukin-8, and related peptides of the chemokine family secreted by gastric epithelial cells, are likely to be important host mediators inducing neutrophil migration to sites of infection. Epithelial IL-8 is upregulated by TNF-alpha and IL-1 and directly by H. pylori strains expressing the CagA phenotype. The extent of mucosal injury may reflect bacterial density, the variability of different strains of H. pylori to induce chemokine expression in epithelial cells and the oxidative burst in neutrophils. Recent evidence from in vivo and in vitro studies shows that CagA+ VacA+ strains of H. pylori are associated with enhanced inflammatory responses and mucosal damage. Defining the specific bacterial mediators of mucosal inflammation will be important in elucidating the role of H. pylori in the pathogenesis of gastroduodenal disease. PMID- 8730258 TI - The role of lipopolysaccharide in Helicobacter pylori pathogenesis. AB - The present review describes the structure, attributes and properties of Helicobacter pylori lipopolysaccharides (LPS), and their potential role in pathogenesis. Although possessing certain attributes similar to those of LPS of other Gram-negative bacteria, H. pylori LPS possess unique biological properties. H. pylori LPS has, in general, low immunological activity and this property may aid the persistence of infection. The O-specific chain of the LPS mimics Lewis blood group antigens in structure. As these antigens are present in the gastric mucosa, the expression of Lewis antigens on the bacterial surface may camouflage the bacterium and aid survival of H. pylori. Alternatively, since autoantibodies against human antral gastric mucosa have been observed in H. pylori-positive patients, the relevance of LPS in the development of autoimmunity in H. pylori associated disease requires further investigation. H. pylori LPS in part mediates the binding of the bacterium to laminin, and interferes with gastric cell receptor-laminin interaction, thereby potentially contributing to the loss of mucosal integrity. In vitro observations of inhibition of sulphated mucin synthesis and stimulation of pepsinogen secretion by LPS suggest new mechanisms for H. pylori-induced mucosal damage. Nevertheless, further in vivo studies are required to support their pathogenic role. PMID- 8730259 TI - Heat shock proteins of Helicobacter pylori. AB - As in any other bacterium, Helicobacter pylori synthesizes two heat shock proteins, the HspA (GroES or Hsp 10 homologue) and the HspB (GroEL or Hsp60 homologue). This article summarizes the present knowledge of genetics, function and the antigenic, immunogenic and protective properties of these two abundant proteins. H. pylori HspA and HspB antigens have vital functions for the bacterium; they share most of the bacterial chaperonin characteristics. However, the unique structure of HspA and its unique capacity to specifically bind nickel ions, strongly suggest an essential role of HspA with regard to the urease metallo-enzyme. The putative role of the H. pylori Hsp antigens in autoimmunity is also addressed. PMID- 8730260 TI - The role of Helicobacter pylori urease in the pathogenesis of gastritis and peptic ulceration. AB - Helicobacter pylori produces a 550 kDa, multimeric, nickel-containing urease that catalyses the hydrolysis of urea to yield ammonia and carbonic acid. The ure gene cluster, comprised of seven genes, encodes the two structural subunits UreA (26.5 kDa) and UreB (60.3 kDa), and five accessory proteins: UreI, UreE, UreF, UreG and UreH. Accessory proteins are required for nickel ion insertion into the apoenzyme. The native protein consists of six copies each of UreA and UreB; two nickel ions are coordinated into each UreB active site. Urease is found in the cytosol, but may also localize on the surface (although this may be an artefact) and elicits a strong serum immunoglobulin response. Urease aids in colonization of the host by neutralizing gastric acid and providing ammonia for bacterial protein synthesis. Host defences are avoided by urease by continuing to neutralize acid locally and by shedding urease, which may be bound by immunoglobulin, from the surface of the bacterium. Host tissues can be damaged directly by the urease-mediated generation of ammonia and indirectly by urease induced stimulation of the inflammatory response, including recruitment of leukocytes and triggering of the oxidative burst in neutrophils. PMID- 8730261 TI - Helicobacter pylori enzymes. AB - Helicobacter pylori exhibits a complex system of enzymes which serve a range of functions, such as colonization, damage of the host epithelium and provision of essential metabolic substrates. Colonization is favoured by urease and by the action on mucus and the mucosal barrier exerted by phospholipases and proteases, although this latter mechanism is controversial. Toxic effects are effected by urease, alcohol dehydrogenase (ADH), phospholipases and proteolytic enzymes. ADH produces acetaldehyde that is toxic to the mucosal cells, while phospholipases induce generation of products such as lysolecithin, which damage the gastric epithelium. Catalase and sodium dismutase of H. pylori are mainly involved in transforming toxic oxygen metabolites to harmless water; they protect the bacterium from the killing effect of neutrophils. Metabolic enzymes (for example, phosphatases, ATPases) are essential for the generation of energy, for synthesis and transport of cell products and for ion fluxes. In addition, they influence cell growth and the expression of virulence factors. PMID- 8730262 TI - Role of vacA and the cagA locus of Helicobacter pylori in human disease. AB - Helicobacter pylori are 'slow' bacteria that may cause disease decades after acquisition. Bacterial pathogenesis often involves features, including conserved genes, shared by many different species. As such, despite its unique niche in the human body, the pathogenesis of H. pylori infection most likely shares mechanisms with other bacteria. In this paper, two genes, vacA and cagA, which appear unique to H. pylori and which may reflect the particular requirements of H. pylori for long-term residence in the human stomach will be discussed. At present the function of these genes for H. pylori is not known yet other characteristics have been defined. PMID- 8730263 TI - Helicobacter pylori exotoxins and gastroduodenal diseases associated with cytotoxic strain infection. AB - This paper describes the characteristics of exotoxins produced by Helicobacter pylori, and in particular the vacuolating toxin (VacA) and the cytotoxin associated protein (CagA). The possible association between infection by strains of certain phenotypic or genomic types and the seriousness of gastroduodenal diseases is discussed. Helicobacter pylori induces various morphological changes in cells in vitro, but only infection by strains which induce cytovacuolation has been studied at present. In its native form, VacA is a protein aggregate made of subunits with a mass of 95 kDa. In vitro it stimulates a cellular v-type ATPase present on the endosomes and creates an acidic environment inside the vacuoles. It also alters in vitro a K(+)-dependent phosphatase activity and could impair the flux of sodium through the cells. Purified VacA causes ulceration in mice; experimental infection in mice with strains which also express the CagA protein causes gastric erosions, vacuolation and epithelial and stromal polymorphonuclear (PMN) cell infiltration. In vivo vacuolation can be observed in gastric cells from patients infected with type I (VacA-CagA positive) H. pylori. CagA is a protein of 128-140 kDa molecular weight, noncytotoxic and highly immunogenic. It is coexpressed in approximately 70% of cytotoxin-producing strains. In CagA positive strain infection, increased levels of interleukin-8 (IL-8) are secreted by the colonized gastric mucosa. Patients infected by cytotoxic strains and/or patients with anti-CagA antibodies are more likely to have active gastritis, and are more likely to develop peptic ulcer or gastric cancer. The different outcomes of infection could be determined by host factors, diet, or by the age at which infection is acquired. PMID- 8730264 TI - Gastrin and acid in relation to Helicobacter pylori. AB - H. pylori infection, both in normal healthy subjects and patients with duodenal ulcer, results in modest elevations of serum gastrin concentrations in the fasting state and quite substantial elevations after a meal or gastrin releasing peptide (GRP) stimulation. Cure of the infection leads to normalization of gastrin homeostasis. Acid secretion in response to a submaximal infusion of GRP is three-fold higher in H. pylori-infected normal subjects and six-fold higher in DU patients than in non-infected controls. These changes also normalize after cure of the infection. H. pylori infection appears to lead to increased basal acid output in some patients with duodenal ulcer while effects on peak acid output to pentagastrin remain under debate. With the possible exception of peak acid output, the abnormalities of gastrin and acid secretion reported for patients with duodenal ulcer are largely a result of infection with H. pylori. PMID- 8730265 TI - The relationships between chronic gastritis and gastric acid secretion. AB - Helicobacter pylori is the main cause of chronic gastritis in humans. Autoimmune mechanisms and Helicobacter heilmannii infection are other causes, both of which are of minor significance in a worldwide perspective. Atrophic gastritis is a quite common late consequence of H. pylori gastritis and will develop on a multifactorial basis, but not in all infected persons. The evolution of atrophic gastritis is a slow and gradually worsening process leading to subtypes, in which the antrum and corpus are affected to dissimilar extent and degree. The distal part of the stomach is the site where the atrophic sequelae (atrophic gastritis and intestinal metaplasia) of H. pylori infection occur most often. A minority of cases develop corpus-limited, or corpus-predominant atrophic gastritis. Along with the worsening of atrophic gastritis, inflammation and density of colonization of the mucosa by H. pylori tend to decrease in grade. In general, the degree of gastric mucosal inflammation, acute and chronic, is positively related to the degree of colonization of the mucosa by H. pylori. Acid secretion and local acidity are factors which modulate the ecology and density of colonization of H. pylori in the stomach, and may thus also modulate the evolution of chronic gastritis into topographically dissimilar subtypes. Acid secretion varies among individuals, this variation being perhaps caused by hereditary differences in parietal cell mass, or by differences in the sensitivity of parietal cells to hormonal or neural stimuli. It is hypothesized that in genuine hypersecretors, H. pylori colonization and subsequent gastritis with atrophic and metaplastic sequelae may be limited to the antrum, while in hyposecretors gastritis predominates in the corpus. In the latter, atrophic gastritis in the corpus then leads to further impairment of acid output. In these cases, H. pylori infection and gastritis may, finally, heal in the antrum, resulting in hypochlorhydria and atrophic gastritis that is limited to, or predominant in the corpus. PMID- 8730266 TI - Gastric metaplasia: its role in duodenal ulceration. AB - There is good evidence to suggest that gastric metaplasia in the proximal duodenum and Helicobacter pylori gastritis are essential requirements for the development of duodenal ulceration in most cases. Gastric metaplasia is most likely to be a defence response or adaptation to excess acid reaching the duodenum. The appearance of gastric-type epithelium over the duodenal villi probably results from substitution by cells migrating from Brunner's gland ducts. These metaplastic foci provide sites for colonization by H. pylori passing through the duodenum; the organisms do not attach to native duodenal epithelial cells. Having colonized the metaplastic areas, H. pylori provokes an active chronic inflammatory response akin to that seen in the gastric mucosa. Active chronic duodenitis leads to a weakening of mucosal defence against acid-peptic attack, and erosion and ulceration may ensue. Healing of ulcers by conventional acid-reducing treatments does not influence the extent of gastric metaplasia, (although there may be some reduction with long-term proton pump inhibitors); nor do such regimens affect the background duodenitis. Only with eradication of H. pylori is there resolution of inflammation, but studies to date indicate that eradication alone has no substantial effect on the prevalence or extent of gastric metaplasia. Nevertheless the elimination of H. pylori appears to remove one of the essential co-factors for duodenal ulceration and the patient can be considered cured, despite the persistence of gastric metaplasia. PMID- 8730267 TI - Vaccination and mucosal responses to Helicobacter pylori infection. AB - H. pylori infection persists for life if not treated, and is responsible for major morbidity and mortality throughout the world. Preventative immunization, once thought to be impossible, is now considered by many to be the only practical approach to large-scale elimination of the bacterium from susceptible populations. High rates of protection have been achieved in the H. felis mouse model, utilizing antigens ranging from whole cells to purified recombinant proteins selected on the basis of their role in pathogenicity. Immunization has also been shown to cure established infection. H. pylori mouse models have been developed and may become the model of choice. Urease remains the favourite antigen but combinations will most likely be required. A priority is to define alternate muscosal adjuvants, as some used in the animal models may be too toxic for use in humans. Also, there is a need to understand the basis of immunization. Why does the natural immune response to H. pylori fail while the artificially stimulated response succeeds? The first important steps towards a vaccine have been made but, given safety issues and regulatory requirements, it may be 5-8 years before the final product becomes available. Over these years antimicrobial resistance is likely to be an increasing problem in the treatment of H. pylori infections. Thus, when the vaccine comes, the time will be ripe for the completely new approach of therapeutic immunization. PMID- 8730268 TI - Local and systemic chemotherapy in the management of periodontal disease: an opinion and review of the concept. AB - Periodontal disease appears to arise from the interaction of pathogenic bacteria with a susceptible host. The main aims of disease management have been to establish a high standard of oral hygiene and to professionally and thoroughly debride the root surface Chemical agents could be considered for both aspects of management. Chemoprevention using supragingivally delivered agents such as chlorhexidine may be questioned for value in the pre-treatment hygiene phase but have well-established efficacy immediately preoperatively and during the post operative weeks. Long-term maintenance use of chlorhexidine is problematic due to local side effects. Antiplaque toothpastes show modest benefits to gingivitis but are not proven to prevent recurrence of periodontitis. Chemotherapy may be directed at subgingival plaque, using antimicrobials, or at the host response using anti-inflammatory agents. Antimicrobials can be locally or systemically delivered. In most cases antimicrobial chemotherapy should be considered adjunctive to mechanical debridement. The advantages of local and systemic chemotherapy must be balanced against the disadvantages and potential side effects of agents. Antimicrobial chemotherapy offers little or no benefit to the treatment of most chronic adult periodontitis patients and should be reserved for the more rapid or refractory types of disease, and after the debridement phase. Despite the large number of studies there are insufficient comparative data to support any one local delivery system or systemic regimen as superior to another. Systemic versus local antimicrobials have not been compared to date. Host response modifying drugs such as non-steriodal anti-inflammatory drugs (NSAIDS) offer the potential to reduce breakdown and promote healing, including bone regeneration. However until more data are available, NSAIDs should not be used in the management of chronic periodontal diseases, there being no specific agent(s) or regimen established for use. Chemotherapy has an important place in the management of chronic periodontal diseases but routine use must be considered as an over prescription of these valuable agents. PMID- 8730269 TI - The prevalence of toothwear in 1007 dental patients. AB - The dental profession is becoming increasingly concerned about unacceptable levels of toothwear and there is an impression that the condition is becoming more prevalent. There is, as yet, little epidemiological evidence to illuminate the question and so this study is therefore intended to provide baseline data on the prevalence of toothwear in a sample of the dental attending population of South East England. A degree of toothwear is normal and progresses throughout life. Toothwear which is acceptable in an older age group would not be so with younger patients. It is therefore necessary to designate threshold values of acceptable levels of toothwear for different age groups to reach realistic conclusions about the prevalence of unacceptable levels of toothwear. Of the 1007 patients examined only nine had completely unworn dentitions. More than 93,500 tooth surfaces were examined and of these 5.1% had wear which exceeded the threshold values. Despite increasing the threshold values for the older age groups from those proposed in an earlier study by Smith & Knight (1984), these patients still had higher levels of unacceptable toothwear. There was a slight tendency for men to have more wear than women, but there was no clinically significant relationship between the number of missing posterior teeth and wear of the anterior teeth. PMID- 8730270 TI - Reduction of microleakage in class II composite resin restorations using retentive pins. AB - The effect on microleakage of Bondent parapulpal pins used in class II composite resin restorations was evaluated in vitro. Bulk and incremental packing and polymerization techniques were compared, using pins anchored at the gingival cavity walls. Evaluation of microleakage was carried out following cyclic load application and an extended thermocycling procedure by the assessment of dye penetration. The results showed that the use of pins at the gingival wall of class II composite resin restorations provided significantly improved marginal sealing. The procedure should, therefore, be regarded as an effective clinical practice. PMID- 8730271 TI - The effect of a vital bleaching technique on enamel surface morphology and the bonding of composite resin to enamel. AB - This study examined the effect of a nightguard vital bleaching procedure on enamel surface morphology and the shear bond strength of a composite resin luting cement to enamel. Extracted human teeth were bleached for 1 week using a vital bleaching product. Control teeth were brushed with a fluoride toothpaste and processed similarly to the bleached teeth, however the bleaching product was substituted with artificial saliva in the night guards. Teeth were stored in artificial saliva for 24 h, 1, 6 or 12 weeks and then examined for any surface changes using light and scanning electron microscopy. The effect of etching surfaces with 37% phosphoric acid was examined at the scanning electron microscope level. The shear bond strength of composite resin luting cement to both buccal and lingual surfaces of bleached and control teeth was determined. Light microscopy investigation suggested the bleaching process resulted in a loss of mineral from enamel which was evident 24 h after bleaching and was sustained following 12 weeks storage in artificial saliva. Scanning electron microscopy showed a definite change in the surface texture of the bleached enamel surface. Acid etching of the bleached enamel surface produced loss of prismatic form and the enamel appeared overetched. The mean shear bond strength between composite resin luting cement and etched enamel tended to be lower for bleached enamel surfaces, however no significant difference in shear bond strength was noted between control and experimental groups. The results of this study suggest that bleaching resulted in changes to the surface and subsurface layers of enamel. Although surface changes were observed in the etched enamel, the shear bond strength of composite resin luting cement to etched bleached enamel appeared to be clinically acceptable. PMID- 8730272 TI - The evaluation of masticatory function with low adhesive colour-developing chewing gum. AB - Many objective methods have been described to evaluate masticatory function. Recently, a new chewing gum system has been reported that utilizes a phloxine sodium bicarbonate reaction and measures a chromatic coordinate as a functional indicator. A low-adhesive colour-developing chewing gum has been developed by selecting an appropriate gum base. This study examined the applicability of the system and reproducibility of results in subjects with different dental states. Tests were performed on 15 natural dentate subjects and 15 complete denture wearers after determination of germane test conditions through a pilot study. The colour was measured by a*, one of the coordinates in the L*a*b* colour system. All subjects could perform the test. The coefficient of variation ranged from 1.87% to 7.90%, averaging 5.15% in the natural dentate subjects and from 1.5% to 17.9%, averaging 9.75% in the complete denture wearers. The 95% confidence interval of a* was 28.76 +/- 1.76 in the natural dentate subjects and 10.05 +/- 2.58 in the complete denture wearers. The results indicated that this material can be useful for both complete denture wearers and subjects with a natural dentition with adequate reproducibility. PMID- 8730273 TI - The strength of a heat-pressed all-ceramic restorative material. AB - A heat-pressed technique (IPS Empress, Ivoclar) has been described to construct single unit crowns, inlay/onlays and veneers using precerammed and precoloured glass-ceramic ingots. The aim of the study was to evaluate the strength of materials obtained using this technique. Strengths were determined by means of shell (disc rupture) tests. Seven groups of 10 specimens each were prepared with different combinations of core, incisal and glaze firing. Shell strength values of these seven groups varied between 167 +/- 18 and 81 +/- 9 MPa. After analysis, results showed that there was no statistical difference between groups when the core porcelain was in tension. However, there was a significant difference between those groups in which the lower surface was either core or incisal porcelain. The component thicknesses of the core and incisal materials did not change the strength of the complete specimen. PMID- 8730274 TI - Patterns of jaw muscle activity during voluntary chewing. AB - Mastication is controlled by central rhythm and burst generators, but whether this control is similar under automatic and voluntary chewing is not clear. Our first aim was to determine whether increased chewing frequency led to a proportional decrease in both the burst duration (the time of muscle activity) and the interburst interval (time of inactivity) of each muscle or to a minimal decrease in the burst duration and a substantial decrease in the interburst interval. Our second aim was to determine whether these two manifestations of control differed between the opening and closing muscles. Electromyograms from digastric, medial pterygoid, masseter and anterior temporalis muscles were obtained during gum chewing by five subjects at four frequencies. A nearly fourfold decrease in total cycle duration (TCD) with increased chewing frequencies was accompanied by minimal changes in burst duration in the digastric muscle but proportional changes in the jaw closing muscles. The onset latency of the digastric muscle as a percentage of TCD decreased and became negative (i.e. occurred before opening started) as the TCD decreased. The onset latencies of the jaw closing muscles remained roughly 10-20% of the TCD for the three lower chewing frequencies. Control of the opening muscle appears to differ from control of the closing muscles. PMID- 8730275 TI - The relationship between jaw elevator muscle surface electromyogram and simulated food resistance during dynamic condition in humans. AB - In six human subjects, electromyograms (EMGs) of the masseter and temporal muscles were recorded bilaterally during experiments in which the subjects made rhythmic open-close movements. The closing phase was counteracted by a variable external force on the mandible. Variables of the force (amplitude, time integral and work) and variables of the corresponding EMG bursts (duration, peak amplitude and time integral) were computed for each open-close cycle. Linear regression analyses were used to determine the strength of the relationship between each EMG variable and each force variable. By step-wise multiple regression analysis the EMG variables predicting the force variables were determined. Although there was a highly significant and positive correlation (P < 0.0005), the average coefficients of linear correlation varied from 0.46 to 0.82. The strongest relationship was observed between the time integrals of the force and EMG in the interval between the onset of the burst and the onset of occlusion. It was suggested that to assess muscle force during chewing, the time integral of EMG bursts should be computed. PMID- 8730276 TI - Failure of dental bridges. I. Age and sex of patients and distribution of replaced and abutment teeth. AB - In this study, 156 dental bridges, constructed in 132 randomly selected patients were evaluated. There was no significant difference between the number of bridges placed in males and the number placed in females. However, the results revealed that the maxillary arch received more prostheses than the mandibular arch. The most-frequently replaced teeth were the mandibular first molars and the least frequently replaced were mandibular canines. Maxillary second premolars, the mandibular second premolars and the mandibular second molars were the most frequent teeth used as abutments. PMID- 8730277 TI - Predictability of jaw muscle pains from surface electromyograms. AB - In seven (88%) of eight healthy subjects, weak to moderate pains were elicited in the masseter muscles through the isometric contractions of maximum voluntary teeth clenching. Integrated surface electromyograms of the right and left masseter muscles were used to quantify the absolute and relative contractile activities of the two muscles. The risk (relative probability) of inducing pain onset in the single masseter muscle generating the larger amount of isometric activity was 2.5 times the risk of eliciting pain onset in the single masseter muscle generating the lesser amount of isometric activity. However, as an aid in the diagnosis of pain onset, the method of masseteric surface electromyography had a false diagnostic ratio of 0.67. PMID- 8730278 TI - Implications for medical genetics of the House of Commons Science and Technology Committee's report on human genetics. PMID- 8730280 TI - Correlation between the development of extracolonic manifestations in FAP patients and mutations beyond codon 1403 in the APC gene. AB - The APC gene was investigated in 31 unrelated polyposis coli families by SSCP analysis and the protein truncation test. Twenty-three germline mutations were identified which gave rise to a variety of different phenotypes. Some of these mutations have already been described; however we report six previously unpublished mutations. Typical disease symptoms were observed in families who harboured mutations between exon 4 (codon 169) and codon 1393 of exon 15. Mutations beyond codon 1403 were associated with more varied phenotype with respect to the development of extracolonic symptoms. In this report we provide support for the notion that there appears to be a correlation between the location of an APC mutation (beyond codon 1403) and extracolonic manifestations of familial adenomatous polyposis. PMID- 8730279 TI - A modifying locus for familial adenomatous polyposis may be present on chromosome 1p35-p36. AB - Mutations of the APC gene cause familial adenomatous polyposis (FAP) in humans and multiple intestinal neoplasia (Min) in laboratory mouse strains. A dominant modifying gene (Mom1), which partially suppresses the min phenotype, has been mapped to mouse chromosome 4. This region is syntenic with human chromosome 1p35 p36. The phospholipase A2 (Pla2s) locus is an excellent candidate for Mom1 and the equivalent human locus PLA2G2A is found on chromosome 1p35. It does not necessarily follow, however, than any modifier of mouse polyposis also influences human disease. In order to test whether a locus on 1p modifies FAP, subjects from 28 FAP families have been typed at microsatellite loci on this chromosome arm. The severity of their duodenal polyposis has also been assessed by endoscopy. Pedigree (lod score) linkage analysis found no evidence of a simple, dominant modifying gene, comparable with the action of Mom1 in inbred mouse strains. Given the more complex genetic and environmental interactions likely to exist in outbred human populations, it is probably more appropriate to use tests which do not specify a mode of inheritance. Using these methods of analysis, the data suggest that a locus on chromosome 1p35-p36 may influence the severity of duodenal FAP. PMID- 8730281 TI - Large scale deletions of the 5q13 region are specific to Werdnig-Hoffmann disease. AB - Spinal muscular atrophy (SMA) is characterised by degeneration of anterior horn cells of the spinal cord and represents the second most common, lethal, autosomal recessive disorder after cystic fibrosis. Based on the criteria of the Internatinal SMA Consortium, childhood SMAs are classified into type I (Werdnig Hoffmann disease), type II (intermediate form), and type III (Kugelberg-Welander disease). Recently, two genes have been found to be associated with SMA. The survival motor neurone gene (SMN) is an SMA determining gene as it is absent in 98.6% of patients. A second gene, XS2G3, or the highly homologous neuronal apoptosis inhibitory protein gene (NAIP) have been found to be more frequently deleted in type I than in the milder forms (types II and III). We investigated the correlation between the clinical phenotype and the genotype at this loci. A total of 106 patients were classified into type I (44), type II (31), and type III (31) and analysed using SMN, markers C212 and C272, and NAIP mapping upstream and downstream from SMN respectively. The combined analysis of all markers showed a large proportion of type I patients (43%) carried deletions of both SMN and its flanking markers (C212/272) and NAIP exon 5), as compared with none of the patients with type II or III SMA. The presence of large scale deletions involving these loci is specific to Werdnig-Hoffman disease (type I) and allows one to predict the severity of the disease in our series. PMID- 8730282 TI - The molecular defect underlying canine fucosidosis. AB - Fucosidosis is a lysosomal storage disease which affects humans and English springer spaniel dogs. The disease is recessively inherited in both species and results from a deficiency of the enzyme alpha-L-fucosidase. We have recently cloned and sequenced the canine fucosidase gene (EMBL sequence admission number X92448 (cDNA) and X92671-X92678 (individual exonic data)). The gene spans 12 kb and consists of eight exons. SSCP based mutation analysis of affected animals was carried out on the coding region of this gene both with exonic primers, and intronic primer pairs for each exon. A 14 base pair deletion of the cDNA was identified at the 3' end of exon 1 in fucosidosis affected animals. Surprisingly, PCR based genomic cloning of DNA from these animals showed an identical deletion in this DNA, ending at the start of intron 1. This change causes a frameshift and, in consequence, 25 novel codons are transcribed in exon 2 before the first of two adjacent premature stop codons is encountered. PMID- 8730283 TI - The dysmorphic human-mouse homology database (DHMHD): an interactive World-Wide Web resource for gene mapping. AB - Genetic mapping and the examination of "candidate genes" for isolating loci associated with clinical syndromes can be greatly accelerated if there is information about where in the genome a particular locus might be situated. Such clues can come from homology to mouse mutants that have been mapped and knowledge of homology between mouse and human chromosomal segments. Further clues can come from chromosome aberrations giving a similar phenotype. However, these clues are often scattered widely in published reports, and even if they are collected together in catalogues or databases there is no rapid way of moving from one data type to another. The Dysmorphic Human and Mouse Homology Database (DHMHD) is designed to ease this data transition. DHMHD comprises detailed information from four separate sources and enables cross referencing through phenotypic and chromosome homology. The DHMHD system is a prototype which is now available online through the World-Wide Web. PMID- 8730284 TI - Complementation analysis in patients with the clinical phenotype of a generalised peroxisomal disorder. AB - The generalised peroxisomal disorders (GPDs) Zellweger syndrome (ZS), neonatal adrenoleucodystrophy (NALD), and infantile Refsum's disease (IRD) are autosomal recessive disorders associated with a failure to assemble mature peroxisomes. We confirmed the diagnosis of a GPD in eight ZS and four IRD patients (GPD1 to GPD12) biochemically by measuring very long chain fatty acids, plasmalogen biosynthesis, and catalase solubility in skin fibroblasts. One further patient (BOX-1) had the clinical phenotype of ZS, but biochemical investigations indicated an isolated deficiency of peroxisomal beta oxidation. To date a total of 10 complementation groups (CGs) for the GPDs and three further CGs for isolated beta oxidation deficiencies have been identified. Most GPD patients have been shown to belong to CG-1 (Baltimore classification); among the rarer groups, CG-4 and CG-8 predominate. We performed somatic cell hybridisation experiments on strains GPD-1 to GPD-12 using plasmalogen biosynthesis as a marker for correction and found that six ZS and three IRD patients, eight of whom were of UK origin, belonged to CG-1. Strain GPD-11, a patient of UK origin with an unusual biochemical phenotype, belonged to CG-8. Strains GPD-10 and GPD-12 were derived from ZS patients of Arabian and Pakistani origin and belonged to the rarer CGs 2 and 7, respectively. Furthermore, complementation analysis using beta oxidation as a marker showed that BOX-1 had an isolated deficiency of the bifunctional protein. PMID- 8730285 TI - Holt-Oram syndrome: a clinical genetic study. AB - A clinical and genetic study of the Holt-Oram syndrome (HOS) has been carried out in the United Kingdom involving 55 cases designated Holt-Oram syndrome, together with their parents and sibs. Data from the clinical assessment of both familial and isolated cases were used to define the HOS phenotype and to outline the spectrum of abnormalities, especially factors affecting severity. Skeletal defects affected the upper limbs exclusively and were bilateral and asymmetrical. They ranged from minor signs such as clinodactyly, limited supination, and sloping shoulders to severe reduction deformities of the upper arm (4.5%). The radial ray was predominantly affected than the right. All affected cases showed evidence of upper limb involvement. Cardiac defects were seen in 95% of familial cases and included both atrial septal defect (ASD, 34%) and ventricular septal defect (VSD, 25%); 39% had only ECG changes. Cardiac involvement ranged from asymptomatic conduction disturbances to multiple structural defects requiring surgery in infancy. Sudden death could be caused by heart block. Inheritance was autosomal dominant with 100% penetrance and no evidence of reduced fitness. Increasing severity occurred in succeeding generations consistent with anticipation. PMID- 8730286 TI - Suggestion of a major gene for familial febrile convulsions mapping to 8q13-21. AB - Febrile convulsions affect 2 to 5% of all children under the age of 5 years. These convulsions probably have a variety of causes, but a genetic component has long been recognised. A large and remarkable family is described in which febrile convulsions appear to result from autosomal dominant inheritance at a single major locus. A gene for febrile convulsions was excluded from regions of previously mapped epilepsy genes and extension of exclusion mapping, using microsatellite markers, to the entire genome implied that a locus on chromosome 8q13-21 may be involved. Linkage analysis of markers on chromosome 8 gave a multipoint lod score of 3.40, maximised over different values of penetrance and phenocopy rate, for linkage between the gene for febrile convulsions and the region flanked by markers D8S553 and D8S279. This lod score was calculated assuming the disease has a penetrance of 60% and a phenocopy rate of 3%. Although there was no indication of linkage other than to markers on chromosome 8, linkage remains suggestive rather than significant because of the maximisation procedure applied. The support for linkage involving a major gene, as opposed to an alternative hypothesis of a complex inheritance pattern, relied upon the assumption of low penetrance. PMID- 8730287 TI - Parents' responses to predictive genetic testing in their children: report of a single case study. AB - There is a widely held view among health professionals that predictive genetic testing of children for late onset diseases is not desirable clinical practice. Yet, little is known about the views of parents, or their responses, to predictive genetic testing in their children. Since such testing is being carried out in some genetic centres, the opportunity was taken to conduct a single case study of the parents of 2 and 4 year old sisters who were tested for the gene for familial adenomatous polyposis. Interviews before testing, after, and 15 months later showed a stable attitude, that parental responsibility included making decisions about such testing, and that the role of health professionals should be one of information giving rather than decision making. These parents had no regrets about having their children tested and reported no changes in their behaviour towards either the child who tested positively or the child who tested negatively. Using standardised scales, mood was found to be within the normal range both before and after testing in the mother and father. This case study is a first step towards systematic empirical studies determining the consequences of acquiescing to parents' requests for genetic testing in their children. PMID- 8730288 TI - Syndromes with lissencephaly. PMID- 8730289 TI - A novel splice site mutation in a Becker muscular dystrophy patient. AB - A Becker muscular dystrophy patient was found to have a single base substitution at the 5' end of intron 54. This single base substitution disrupts the invariant GT dinucleotide within the 5' donor splice site and was shown to cause an out of frame deletion of exon 54 during mRNA processing. This is predicted to produce a truncated dystrophin protein which is more consistent with a DMD phenotype. However, small quantities of normal mRNA are also transcribed and these are sufficient to produce a reduced amount of normal molecular weight dystrophin and give rise to a milder BMD phenotype. This indicates that a single base substitution at an invariant dinucleotide of the splice site consensus sequence may still allow read through of the message and allow the production of some normal protein. This shows that there are a greater number of possible intronic mutations that can lead to a mild phenotype and it also underlines the importance of performing cDNA analysis when screening for small gene alterations in the BMD patient population. PMID- 8730290 TI - Phenotypic expression in von Hippel-Lindau disease: correlations with germline VHL gene mutations. AB - Von Hippel-Lindau disease is an autosomal dominantly inherited familial cancer syndrome predisposing to retinal and central nervous system haemangioblastomas, renal cell carcinoma, and phaeochromocytoma. VHL disease shows variable expression and interfamilial differences in predisposition to phaeochromocytoma. In a previous study of 65 VHL kindreds with defined VHL mutations we detected significant differences between VHL families with and without phaeochromocytoma such that missense mutations were more common and large deletions or protein truncating mutations less frequent in phaeochromocytoma positive families. To investigate the significance and cause of this association further, we studied 138 VHL kindreds for germline mutations and calculated the age related tumour risks for different classes of VHL gene mutations. Using SSCP, heteroduplex and Southern analysis we identified a germline VHL gene mutation in 101 families (73%). Direct sequencing of the VHL coding region further increased the mutation detection rate to 81%. In addition to precise presymptomatic diagnosis, identification of a VHL gene mutation can provide an indication of the likely phenotype. We found that large deletions and mutations predicted to cause a truncated protein were associated with a lower risk of phaeochromocytoma (6% and 9% at 30 and 50 years, respectively) than missense mutations (40% and 59%, respectively) and that missense mutations at codon 167 were associated with a high risk of phaeochromocytoma (53% and 82% at ages 30 and 50 years). Cumulative probabilities of renal cell carcinoma did not differ between the two groups (deletion/ truncation mutations: 8% and 60%, and missense mutations: 10% and 64% at ages 30 and 50 years, respectively). Age related risks for haemangioblastoma were similar in the two mutation groups, with the age related risks of cerebellar haemangioblastoma slightly less (35% and 64% v 38% and 75% at ages 30 and 50 years) and retinal haemangioblastoma slightly higher (45% and 72% v 37% and 64% at ages 30 and 50 years) in the missense mutation group than in the deletion/protein truncation group. These results provide valuable data for counselling VHL families and indicate that specific VHL mutations may be associated with different tumour susceptibility risks. There was no evidence of a generalised increase in age related tumour risks for missense mutations, suggesting that missense mutations predisposing to phaeochromocytoma have tissue specific effects, possibly because the VHL protein has several functions, the importance of which varies from tissue to tissue, or because the proteins which interact with VHL differ between different tissues. PMID- 8730291 TI - Familial cluster of ovarian small cell carcinoma: a new mendelian entity? AB - We report a pedigree in which three sisters had a particular type of ovarian cancer, small cell carcinoma of the hypercalcaemic type. This rare type of ovarian carcinoma is now well characterised by clinical and pathological findings and is well distinguished from other ovarian epithelial tumours and ovarian germ cell tumours. The occurrence of this rare type of cancer in several members of the same family and the existence of four other similar published observations raises the question of the genetic determination of this kind of tumour. PMID- 8730292 TI - The angiotensin-I converting enzyme (ACE) gene I/D polymorphism and ACE levels in Pima Indians. AB - An insertion/deletion (I/D) polymorphism of the angiotensin converting enzyme (ACE) gene is associated with plasma ACE levels in white populations. The occurrence of the I/D polymorphism and relationship to ACE levels was examined in a Pima Indian group (n = 305). The frequency of the D allele was lower in Pimas than whites (0.29 v 0.52 respectively). ACE levels were significantly associated with genotype in both groups (p = 0.0001), which accounted for 6.5% of the variation in ACE levels in Pimas and 18% in whites. The association of the I/D polymorphism with ACE levels confirms the relationship across ethnic groups. The low frequency of the D allele in Pima Indians shows that ethnic differences should be accounted for when studying the ACE gene. PMID- 8730293 TI - Mosaicism for the fragile X syndrome full mutation and deletions within the CGG repeat of the FMR1 gene. AB - The main mutation responsible for the fragile X syndrome is the expansion of an untranslated CGG repeat in the first exon of the FMR1 gene, associated with the hypermethylation of the proximal CpG island and the CGG repeat region, and repression of transcription of FMR1. Fragile X syndrome mosaicism has been described as the coexistence of the full mutation and the permutation. We present here two cases of mosaicism for the full mutation in the FMR1 gene and deletions involving the CGG repeat region. In one case the deletion removed 113 bp proximal to the CGG repeat and part of the repeat itself, leaving 30 pure repeats, and representing 17% of lymphocytes of the patient. The 5' breakpoint of this deletion falls outside the putative hotspot for deletions in the CGG region of FMR1. In the second case the deleted region only involved the CGG sequence (leaving 15 pure repeats), with normal sequences flanking the repeat; this deleted ("normal") FMR1 was estimated to be in about 31% of blood lymphocytes. This second case can be considered a true regression of the CGG FMR1 expansion to a normal sized allele, although in mosaic form. PMID- 8730294 TI - Clinical, enzymatic, and molecular characterisation of a Portuguese family with a chronic form of GM2-gangliosidosis B1 variant. AB - Mutations in the hexosaminidase A gene (HEXA) causing the B1 variant of GM2 gangliosidosis result in the presence of a mutant enzyme protein with a catalytically defective alpha subunit. A rare and panethnically distributed mutation, transition G533A (Arg178His), is known to be a common allele among Portuguese patients with the subacute phenotype. We now report the presence of an Arg178His allele in three Portuguese sibs with a chronic form of the disease, who carry the transition G755A (Arg252His) on the second allele. This novel mutation is the first B1 allele to be associated with an adult phenotype. PMID- 8730295 TI - "Pure" partial trisomy 4q25-qter owing to a de novo 4;22 translocation. AB - A girl with a new de novo translocation of 4q onto the short arm of acrocentric 22 is reported as a case of "pure" partial trisomy 4q. Her karyotype was 46, XX, 22, +mar, t(4;22)(q25-->qter;p11) identified by Giemsa staining and FISH. Comparison of the proband with previously reported cases of "pure" partial trisomy 4q showed the main clinical features to be growth retardation, psychomotor retardation, microcephaly, large, low set, malformed ears, prominent nasal bridge, ptosis and epicanthus. PMID- 8730296 TI - Genetics in primary care. Report on workshop of EC Concerted Action on Genetics Services in Europe (CAGSE) in association with the Royal College of GP Spring Meeting, Blackpool, UK, 28 April 1995. PMID- 8730297 TI - Renal and urological tract malformations caused by a 22q11 deletion. PMID- 8730299 TI - Viral hepatitis: control, seroepidemiology and surveillance. PMID- 8730298 TI - Unstable mutation in incontinentia pigmenti? PMID- 8730300 TI - C-reactive protein and haptoglobin in the evaluation of a community-based malaria control programme. AB - When cross-sectional surveys are used to evaluate malaria intervention programmes in the community, the prevalence of morbidity is difficult to assess because of the fluctuating nature of malarial fever. We have therefore investigated the impact of bed net usage on 2 surrogate markers of malarial morbidity: (i) elevated C-reactive protein (CRP) (> 8 mg/L) plus detectable parasitaemia, as an indicator of malaria-induced acute-phase response; and (ii) reduced haptoglobin levels (< 180 mg/L), which in this population indicates malaria-induced intravascular haemolysis. Among 1505 Gambian children 1-5 years old, examined on a single occasion at the end of the malarial transmission season, 5% had parasitaemia plus fever, while 24% had parasitaemia plus elevated CRP, and 35% had low haptoglobin. The proportion of children who had parasitaemia plus elevated CRP was significantly lower among those who had slept under insecticide treated bed nets than among those who did not use a bed net (16% vs. 34%, P < 0.003), and the proportion with low haptoglobin differed similarly (24% vs. 49%, P < 0.003). Use of an untreated bed net had a weaker effect on both indices (22% had parasitaemia plus elevated CRP, 34% had low haptoglobin). CRP and haptoglobin are simple and inexpensive to measure in large numbers of people, and these results suggest that they could be useful for the assessment of malaria intervention programmes. PMID- 8730301 TI - Malaria: even more chronic in nature than previously thought; evidence for subpatent parasitaemia detectable by the polymerase chain reaction. AB - In high endemicity areas, malaria is a chronic disease: examination of blood films reveals that up to half of the population, particularly children, harbour parasites at any one given time. The parasitological status of the remainder was addressed using the polymerase chain reaction, a technique 100 to 1000 times more sensitive than microscopy, on a series of samples from Dielmo, a holoendemic area of Senegal. Two-thirds of the microscopically negative individuals were found to harbour subpatent levels of Plasmodium falciparum, suggesting that more than 90% of the exposed population at any one time, i.e. in a cross-sectional survey, are chronically infected. This also means that the range of parasite loads harboured by humans with various degrees of exposure is remarkably large, probably reflecting a large range of effectiveness of the defence mechanisms against malaria parasites, none of which is fully efficient. PMID- 8730302 TI - Measurement by membrane feeding of reduction in Plasmodium falciparum transmission induced by endemic sera. AB - The standard laboratory test for reduction in malaria transmission is based on the measurement of oocyst numbers in mosquitoes fed on blood meals containing test and control sera. Interpretation of the results, however, is often hampered by the large variation in numbers of infected mosquitoes and oocysts. The objective of this study was to compare 3 measures for the assessment of transmission reduction (so-called R values) and to define the experimental criteria that allow interpretation of the results. To determine variability in R values of control sera, a replicate experiment was performed with 10 non-endemic sera of Dutch blood donors. Furthermore, 2 measures for calculation of transmission reduction were compared in a triplicate experiment using Plasmodium falciparum, Anopheles gambiae and malaria endemic sera. Calculations using the geometric mean of Williams are currently used to identify blocking and non blocking sera. However, calculations using log-transformed data could distinguish more gradual levels of transmission reduction activity by endemic sera--i.e. blocking, reducing and non-blocking activity. Grading of transmission reduction activity is important for epidemiological studies on transmission immunity and for validation of future transmission-blocking vaccines. PMID- 8730303 TI - The late biting habit of parous Anopheles mosquitoes and pre-bedtime exposure of humans to infective female mosquitoes. AB - Using the all-night landing catch method (18:00-06:00) we showed, for Anopheles gambiae in Sierra Leone and A. punctulatus in Papua New Guinea, that parous females have a tendency to bite later than nulliparous ones. The biting habit of sporozoite-infected A. punctulatus was also investigated. The sporozoite rates for Plasmodium falciparum and P. vivax were 1.8 and 1.4% respectively, but only one (1.3%) of 76 females infected with P. falciparum was caught between 18:00 and 21:00. A significantly higher proportion (11.6%) of mosquitoes infected with P. vivax was caught in the same period. The late biting habit of mosquitoes infected with P. falciparum is discussed in relation to the differential biting habits of parous and nulliparous females. We conclude with a hypothesis that, in areas where Anopheles mosquitoes have a late-biting cycle and low parous rate, exposure to mosquitoes infected with P. falciparum during the pre-bedtime period (18:00 22:00) is very low. This hypothesis could explain why insecticide-treated bed nets protect children better in areas of seasonal transmission, where nulliparous females tend to predominate, than in areas of perennial transmission, where parous females are usually more numerous. The same hypothesis is compatible with the finding in Papua New Guinea that insecticide-impregnated bed nets are more protective against P. falciparum than against P. vivax malaria. PMID- 8730304 TI - Descriptive epidemiology of lymphatic filariasis in Ghana. AB - A national filariasis survey was conducted to determine the prevalence and distribution of Wuchereria bancrofti microfilaraemia and clinical disease associated with lymphatic filariasis in Ghana. Multi-stage systematic sampling was used to select households to be examined in all 10 administrative regions of the country. The results showed a high prevalence of microfilaraemia and disease with considerable regional variation in all the quantities investigated. In general, the disease was more prevalent in the northern guinea savannah and the southern coastal savannah, while the middle forest belt was relatively free. Even within the regions, there was considerable variation in the distribution of the disease. The disease is of potentially major public health significance, particularly in the northern and coastal savannahs, and requires the design and implementation of a control programme. PMID- 8730305 TI - Seroprevalence of five arboviruses in Zebu cattle in the Central African Republic. AB - Sequential blood samples taken from 237 adult and 147 young Zebu cattle in the Central African Republic (CAR) were examined by enzyme-linked immunosorbent assay for the presence of antibodies to Rift Valley Fever (RVF), Dugbe (DUG), Crimean Congo haemorrhagic fever (CCHF), Chikungunya (CHIK) and Wesselsbron (WSL) viruses. Antibodies to DUG and CCHF viruses were detected in 70% and 61% respectively, of the sera from adult cattle, probably due to common antigens. This hypothesis is supported by the fact that CCHF virus was isolated only 3 times from ticks, whereas DUG virus was isolated from almost all ticks. Ticks of the genus Hyalorama, the most important vectors of CCHF, are rare on Zebus in CAR. CCHF virus has a limited role in local human pathology, probably because its distribution is limited by the scarcity of its main vector and the immunological barrier resulting from the presence of antibodies to DUG virus. Antibodies to RVF virus were found in about 8% of adult cattle. Zebus do not play an important role in the spread of CHIK and WSL viruses among humans; these viruses were found in only one and 5, respectively, of the 183 cattle examined. PMID- 8730306 TI - Acridine orange fluorescence techniques as alternatives to traditional Giemsa staining for the diagnosis of malaria in developing countries. AB - Traditional Giemsa-stained thick blood films were compared with 2 fluorescence microscopy techniques, acridine orange (AO) staining of thin blood films and the quantitative buffy coat (QBC) method, for the microscopical diagnosis of malaria. Of 200 samples examined, 141 were positive by Giemsa staining, 146 by AO and 137 by QBC. Overall sensitivities for the 2 fluorescence techniques compared to Giemsa staining were good: AO 97.9% and QBC 93.6%. However, with parasitaemias < 100/microL the QBC sensitivity fell to 41.7% whereas that of AO was 83.3%. Both AO and QBC were unable to differentiate accurately between individual malaria species. We conclude that the QBC technique alone cannot replace Giemsa-stained thick blood films for most purposes in an African setting. However, apart from species differentiation, the AO method is an appropriate technique for the laboratory diagnosis of malaria in developing countries. PMID- 8730307 TI - Immunoglobulin gene polymerase chain reaction to distinguish hyperreactive malarial splenomegaly from 'African' chronic lymphocytic leukaemia and splenic lymphoma. AB - Hyperreactive malarial splenomegaly (HMS) is found in geographical association with B cell lymphoproliferative disorders such as 'African' chronic lymphocytic leukaemia (CLL) and splenic lymphoma with villous lymphocytes (SLVL). It is sometimes not easy to make a differential clinical diagnosis between these conditions. We have previously used Southern blotting as a definitive method for the diagnosis of monoclonal lymphoproliferation in these disorders, but this is expensive, lengthy and technically difficult. In the present paper we have compared Southern blotting with polymerase chain reaction (PCR) amplification of the immunoglobulin heavy chain gene. We found an excellent correlation between the 2 methods in demonstrating monoclonal populations of lymphocytes in patients with a clinical diagnosis of CLL or SLVL. We have further demonstrated monoclonality in a patient who could not be classified as CLL or SLVL on clinical criteria alone. In contrast, patients with well defined HMS or with non-B cell proliferations all showed polyclonal rearrangements. We propose that the immunoglobulin gene PCR is a useful tool for the investigation of tropical splenomegaly of uncertain origin. PMID- 8730308 TI - Nested polymerase chain reaction for detection of Plasmodium falciparum infection in Malaysia. PMID- 8730309 TI - Clinical diagnosis of malaria: can the patients help us improve? PMID- 8730310 TI - Antigen detection assay to monitor the efficacy of praziquantel for treatment of Paragonimus westermani infections. PMID- 8730311 TI - High plasma levels of nitrogen oxides are associated with severe disease and correlate with rapid parasitological and clinical cure in Plasmodium falciparum malaria. AB - Plasma levels of nitrogen oxide (NO), neopterin and C-reactive protein (CRP) were compared in 3 groups of Gabonese patients with Plasmodium falciparum malaria before and after therapy: adults with uncomplicated malaria, children with uncomplicated malaria, and children with severe malaria. Plasma levels of all 3 molecules were significantly higher in severe malaria than in uncomplicated malaria. High levels of neopterin and CRP during the acute phase of malaria significantly correlated with slow parasitological and clinical cure after therapy. In contrast, high NO plasma levels during the acute phase of malaria predicted accelerated cure. These findings provide further evidence for the protective role of NO in malaria. However, as NO levels were highest in severe disease, overproduction may be harmful for the patients. PMID- 8730312 TI - Schistosomiasis mansoni in Kenya: relationship between infection and anaemia in schoolchildren at the community level. AB - Haematological surveys were carried out in 3 schools in 2 areas where Schistosoma mansoni is endemic in Machakos District, Kenya, before and after a treatment campaign using praziquantel. Earlier clinical impressions of differences in the levels of anaemia between the 2 areas were not confirmed. Although individual haemoglobin levels and haematocrits often fell below international norms, significant anaemia with abnormal red blood cell morphology was rare (< 5%), but varied between schools. Altitude could have accounted for some of these differences, but other factors, including diet and parasitism, were involved. Anaemia was associated with splenomegaly and, to a lesser extent, hepatosplenomegaly. Epidemic malaria (mainly Plasmodium falciparum) appeared to be the main cause of parasite-induced anaemia. There was no significant association with the scarce hookworm infections (mainly Necator americanus); nor did the much commoner S. mansoni cause severe anaemia at the community level, but haemoglobin levels dropped as its intensity increased. Treatment with praziquantel eliminated this trend except in a few subjects with splenomegaly alone (probably due to malaria) or with schistosomal hepatosplenic disease. Possible pathogenic mechanisms are reviewed, including the consumption of red blood cells by adult schistosomes as a possible cause of anaemia. PMID- 8730313 TI - Occurrence of living adult Wuchereria bancrofti in the scrotal area of men with microfilaraemia. AB - To determine the frequency with which living adult Wuchereria bancrofti can be detected by ultrasound in the scrotal area of men with filarial infection, we used a 7.5 MHz transducer to perform weekly ultrasound examinations on 100 microfilaraemic men (18-34 years old) from Greater Recife, Brazil. The peculiar pattern of movement that characterizes the adult worm image on ultrasound (the filaria dance sign) was detected in the lymphatic vessels of the spermatic cord in 80 men (bilaterally in 29 men). Among 20 men with no filaria dance sign, the geometric mean microfilarial density was 68/mL, compared with 238/mL and 775/mL among those with unilateral and bilateral filaria dance signs, respectively (P = 0.0001). The lymphatic vessels of the spermatic cord appear to be a common, and perhaps the principal, site of adult W. bancrofti in men with asymptomatic microfilaraemia. Studies are needed to define the relationship between the presence of filarial worms in the scrotal area and the development of filaria associated morbidity. PMID- 8730314 TI - Meningitis caused by Cryptococcus neoformans var. gattii and var. neoformans in Papua New Guinea. AB - Eleven cases of cryptococcal meningitis were diagnosed and biotyped from September 1991 to August 1992 in Papua New Guinea (PNG). Seven isolates were Cryptococcus neoformans var. gattii from paediatric and adult patients, one with diabetes mellitus and 4 were C. neoformans var. neoformans from adults, of whom 2 had human immunodeficiency virus type 1 (HIV-1) infection, and one each had tuberculosis and Plasmodium vivax malaria. Significant clinical findings were headache, fever, meningism, vomiting, photophobia, papilloedema and cranial nerve lesions. Five patients (45.5%) died; 3 of these were adults with var. gattii and 2 were men with both var. neoformans and HIV-1 infections. This prospective tropical study documents the emergence of C. neoformans var. neoformans in patients with HIV-1 infection in a country where previously var. gattii had predominated in the immunocompetent. There has been no earlier report of cryptococcosis in an HIV-1 seropositive patient in PNG. Despite presumed exposure to both varieties of C. neoformans, var. gattii infections had been most frequent. As HIV-1 spreads, the proportion of hosts infected with var. neoformans may rise. The course of meningitis caused by the 2 varieties of C. neoformans may differ, with mortality in the tropics remaining particularly high. In PNG the environmental source of C. neoformans remains elusive. PMID- 8730315 TI - Multiple dose pharmacokinetics of oral artemisinin and comparison of its efficacy with that of oral artesunate in falciparum malaria patients. AB - The study compared the clinical efficacy and safety of oral artemisinin and oral artesunate as well as artemisinin pharmacokinetics during and after resolution of falciparum malaria. Forty adults with symptomatic falciparum malaria were allocated at random to treatment with either oral artemisinin (500 mg single dose on day 1 followed by 250 mg twice daily for 4 d and then another 500 mg single dose on day 6) or with oral artesunate (100 mg single dose on day 1 followed by 50 mg twice daily for 5 d). Patients were admitted to hospital at the Kibaha Designated District Hospital, Kibaha, Tanzania for the duration of treatment. The patients were seen once weekly for 3 more weeks. The time to parasite clearance (PCT) after oral artesunate (26.4 +/- 3.6 h) was shorter (P = 0.002) than after artemisinin (31 +/- 3.6 h). The fever subsidence time (FST) after oral artesunate (18.9 +/- 4.0 h) was also shorter (P = 0.04) than after artemisinin (21.8 +/- 4.6 h). Parasites were detected in 4 (20%) and 7 (35%) patients after completing treatment with artesunate and artemisinin respectively. In these patients the parasitaemia reappeared at the 3rd or 4th week of follow-up. Standard haematology, blood biochemistry and urinalysis, performed before drug intake and again on days 6 and 14, were normal. No clinical abnormality was observed during the study period. Artemisinin plasma concentrations, determined by high performance liquid chromatography with post-column derivatization and detection by ultraviolet light, were followed up to 8 h after drug administration on days 1 and 6. Artemisinin absorption was rapid, the maximum plasma concentrations (Cmax) being attained at about 3 h. Artemisinin areas under the plasma concentration time curve (AUC) and the Cmax values were about 6 times higher after the first dose on day 1 than on day 6. This decrease in artemisinin plasma concentration is suggestive of an increase in metabolic capacity due to pronounced autoinduction. PMID- 8730316 TI - Validity of mother's history regarding antimalarial drug use in Malawian children under five years old. AB - History obtained from parents and carers is an important, and often the only, source of information for health workers treating children for malaria, but its validity has not been well evaluated. At 2 hospitals in Malawi, we obtained malaria treatment histories from mothers of 973 ill children reported to have had fever as part of the illness. Urine samples were collected from 755 of the 973 children (78%). Of the 755, 457 (61%) were reported to have received some kind of treatment. Among those who reportedly received treatment, 79 (17%) were said to have received chloroquine and 23 (5%) a sulphonamide-containing medicine; however, when urine specimens were tested for antimalarial drugs, chloroquine was found in 182 specimens (40%) and a sulphonamide in 148 (32%). Among urine specimens collected from 291 children who were reported to have received no treatment (no report was recorded for 7 children), chloroquine was detected in 56 (19%) and a sulphonamide in 44 (15%). Although not statistically significant, mothers often reported a child as not having received an antimalarial drug if the child was younger than 12 months or had been sick for more than 3 d. The mothers' information regarding home treatment of fever in children was highly inaccurate. Malaria treatment histories, whether collected at health facilities or in surveys of knowledge, attitudes, and practices, must be interpreted with caution. PMID- 8730317 TI - Mass diethylcarbamazine chemotherapy for control of bancroftian filariasis: comparative efficacy of standard treatment and two semi-annual single-dose treatments. AB - The efficacy of 2 strategies for control of bancroftian filariasis using mass diethylcarbamazine (DEC) chemotherapy was evaluated and compared in 2 endemic communities in Tanzania with pre-treatment microfilarial (mf) prevalences of 28.5% and 17.7%, and mf geometric mean intensities (GMI) of 588 mf/mL and 251 mf/mL, respectively. All individuals in the first community were offered DEC treatment with 6 mg/kg body weight given daily for 12 d (standard treatment). The second community was offered DEC treatment with 2 single doses of 6 mg/kg body weight given with an interval of 6 months (semi-annual single-dose treatment). Among those who were microfilaraemic before treatment, the mf clearance rates were 51.2% and 36.0%, and the mf GMIs were reduced by 98.6% and 92.2% one year after the start of the standard and the semi-annual regimens, respectively. At community level, the standard strategy and the semi-annual strategy reduced the mf prevalences to 15.1% and 11.6% (reductions of 47.0% and 34.5%) and the mf GMIs to 112 mf/mL and 102 mf/mL (reductions of 81.0% and 59.4%, respectively) one year after start of treatment. Both regimens resulted in remarkable improvements in small hydroceles among males presenting this condition before treatment. The lower efficacy of the semi-annual single-dose treatment in relation to the standard treatment in reducing microfilaraemias might be compensated for by continuing semi-annual treatments for a slightly longer period of time. Considering that the semi-annual treatment is easy to administer and more acceptable to the treated individuals, it may in the long run be a more feasible strategy for mass DEC chemotherapy than the standard treatment. PMID- 8730318 TI - Mass diethylcarbamazine chemotherapy for control of bancroftian filariasis through community participation: comparative efficacy of a low monthly dose and medicated salt. AB - The efficacy of 2 strategies for the control of bancroftian filariasis using diethylcarbamazine (DEC) mass chemotherapy delivered through community participation was evaluated and compared in 2 endemic communities in Tanzania with pre-treatment microfilarial (mf) prevalences of 34.7% and 31.0%, and mf geometric mean intensities (GMI) of 1122 mf/mL and 933 mf/mL, respectively. In the first community, all individuals aged > or = 1 year were offered treatment for one year with a low monthly dose (50 mg DEC to children aged < 15 years and 100 mg DEC to adults aged > or = 15 years; given independently of body weight), and in the second community all households were offered 0.33% w/w DEC-medicated cooking salt for one year. Both treatment strategies resulted in dramatic reductions in the mf loads. Among those microfilaraemic before treatment, the low monthly dose and the DEC-medicated salt gave mf clearance rates of 55.3% and 92.1%, respectively, and the pre-treatment mf GMIs were reduced by 99.4% and 99.9%, respectively, one year after starting treatment. At community level, the mf prevalences were reduced to 15.8% and 2.4% (reductions of 54.5% and 92.3%) and the mf GMIs were reduced to 100 mf/mL and 32 mf/mL (reductions of 91.1% and 96.6%), one year after starting treatment with the low monthly dose and DEC medicated salt respectively. Males with hydrocele before treatment improved remarkably one year after the start of treatment. Since both strategies were simple to administer and well accepted by the communities, they appear highly feasible for integration into large scale control programmes based on community participation. PMID- 8730319 TI - Disruption of erythrocyte rosettes and agglutination of erythrocytes infected with Plasmodium falciparum by the sera of Papua New Guineans. AB - People living in areas endemic for Plasmodium falciparum develop humoral responses which may contribute to protection against clinical disease but the specificity of such protective antibody responses remains to be defined. Antibodies disrupting erythrocyte rosettes have been associated with protection against cerebral malaria, and antibodies agglutinating infected erythrocytes with reduced episodes of clinical disease. We have studied the capacity of serum from Papua New Guinean adults and children with a spectrum of malaria exposure, including children and adults at the time of clinical disease, to disrupt erythrocyte rosettes and cause agglutination of infected erythrocytes. Using a single parasite isolate, almost all sera from adults from highly endemic areas agglutinated infected erythrocytes, and the majority disrupted rosettes, in some cases at greater titres than hitherto described. There was a correlation between rosette disruption and agglutination in highly exposed adults. Rosette disrupting antibodies were equally frequent in children with cerebral and uncomplicated malaria. Antibodies causing rosette disruption were frequent only in adults with a long history of malarial exposure. Rosette disrupting antibodies do not appear to protect Papua New Guinean children or adults against cerebral malaria. PMID- 8730320 TI - Impaired antibody responses and loss of reactivity to Onchocerca volvulus antigens by HIV-seropositive onchocerciasis patients. AB - The impact of concomitant human immunodeficiency virus (HIV) infection on the antibody response of onchocerciasis patients to Onchocerca volvulus antigens (OvAg) was studied by Western blotting and enzyme linked immunosorbent assay (ELISA). Immunoglobulin G (IgG) antibodies in sera from 45 HIV-sero-positive O. volvulus microfilariae (mf) carriers (HIV+/Ov+) recognized significantly fewer distinct O. volvulus antigenic bands, and responded less frequently to all detected bands compared to sera from 61 matched HIV-seronegative mf carriers (HIV /Ov+). 29% of 31 follow-up sera from the HIV+/Ov+ patients failed to react to many of the antigenic bands recognized by initial sera from the same patients. Among 4 HIV+/Ov+ persons examined for total CD4+ cells, loss of reactivity corresponded with low CD4+ total cell counts. In an OvAg ELISA, sera from the HIV+/Ov+ individuals had significantly lower IgG+IgM antibody levels than sera from the HIV-/Ov+ persons, and the sensitivity of the assay was 87% for the HIV+/Ov+ subjects compared to 100% for those who were HIV-/Ov+. It is concluded that HIV-infected onchocerciasis patients exhibit significantly impaired antibody responses to O. volvulus antigens, and tend to lose their reactivity to these antigens over time due to immune response abnormalities caused by the concomitant HIV infection. PMID- 8730321 TI - Tuberculosis mortality and HIV infection. PMID- 8730322 TI - Guinea Bissau: association of chloroquine resistance of Plasmodium falciparum with the Tyr86 allele of the multiple drug-resistance gene Pfmdr1. PMID- 8730323 TI - Do you clean or contaminate your bronchoscope? PMID- 8730324 TI - Inhalers and nebulizers: which to choose and why. AB - It is obvious that many factors should be considered when an inhaler is prescribed. Based upon the information discussed above, a rational inhaler strategy could be as follows: (1) Children < or = 5 years and elderly patients are prescribed a spacer with a valve system (and a face mask for the children) for the delivery of all drugs. When they are severely obstructed, some may need a nebulizer. If the patient cannot be taught the correct use of a spacer, a nebulizer should be prescribed. (2) Children > or = 5 years and adults are prescribed a spacer or a Turbuhaler for the administration of inhaled corticosteroids and a dry powder inhaler (preferably multiple dose) or a breath actuated MDI for other drugs. If these alternatives are not available or the patient prefers, a conventional MDI can be used (preferably not for other corticosteroids than fluticasone propionate) provided that careful tuition is given. Fluticasone dipropionate may be given by DPI, Spacer or MDI. (3) Nebulizers are mainly reserved for severe acute attacks of bronchoconstriction. With this approach, most patients can be taught effective inhaler use with a minimum of instructional time. Finally, it must always be remembered to consider the patient's wish, since prescription of an inhaler which the physician likes but the patient does not is likely to reduce compliance. PMID- 8730325 TI - Different cytokine patterns in bronchial biopsies in asthma and chronic bronchitis. AB - Bronchial biopsies have made possible the detailed study of the pathology of the airways of humans with respiratory disease. Much data has been accumulated on asthmatics or normal controls but much less is known about chronic bronchitics. The aim of this study was to characterize the cellular and cytokine pattern seen in chronic bronchitics and to compare these with control and asthmatic subjects. The patients were also characterized clinically. In this study, immunocytochemistry on cryostat sections from bronchial biopsies were used to determine the level of inflammatory cells and cells of the immune system as well as the pattern of cytokines. This study revealed a distinct cellular and cytokine pattern for each of the three different patient groups, although the diversity of the cytokines analysed was limited by the size of the biopsies. In the inflammatory infiltrate of patients with asthma, CD4+ T-cells and eosinophils were the most prominent cell types discerned. All of the expected cytokines such as IL-1, TNF-alpha, IL-4, IL-5 and IFN-gamma were found. In contrast, the emphasis in chronic bronchitic patients was quite different. The predominant cell types were macrophages, neutrophils, mast cells and CD8+ T-cells, but eosinophils were also abundant. In addition, IL-4 and TNF-alpha were the only cytokines present of those tested. PMID- 8730326 TI - How is sputum microbiology used in general practice? PMID- 8730327 TI - Factors associated with tuberculin reactivity among children in United Arab Emirates. AB - A cross-sectional survey of tuberculin skin reactivity was conducted in Al-Ain, United Arab Emirates (U.A.E.) between January and June 1994, to find out the prevalence rate of tuberculosis infection. A pre-designed questionnaire was used to collect details of BCG scar, age, sex, residence area, nationality, education, type of house, number of rooms, family size and household contact history of tuberculosis. A total of 785 students were screened, of whom 547 gave a history of BCG vaccination in the past and 238 were BCG-negative. Among BCG-negative children aged 5-11 years and 12-15 years, only 6.5% and 9.3%, respectively, had a positive Mantoux reaction--a rate lower than most Third World countries, but higher than developed countries where under 2% of children are tuberculin reactors. A general linear model with positive Mantoux reaction as the dependent variable was fitted to the data to examine the joint effect of age, sex, residential area, number of rooms at home, family size and BCG vaccine history. The Mantoux reaction was entered as positive and a number of statistically significant associations were found between positive Mantoux test > 10 mm and: age (P = 0.0018); sex (P = 0.0281); residential area (P < 0.0001); number of rooms (P = 0.0017); and BCG vaccine history (P < 0.0001). However, family size did not have any statistical effect on tuberculin testing (Mantoux test > 10 mm). The prevalence (8%) in the 5-14 years age group puts U.A.E. between low (2%) and middle (14%) prevalence countries, according to the classification of the International Union Against Tuberculosis. This calls for continuation of free treatment of active cases and increased efforts towards screening of contacts. PMID- 8730328 TI - Tumour necrosis factor, interleukin-1 and adenosine deaminase in tuberculous pleural effusion. AB - Tumour necrosis factor (TNF) and interleukin-1 (IL-1) are powerful mediators with a key role in inflammation. This study was undertaken to study the presence of TNF and IL-1 in tuberculous effusion where there is marked inflammation and where examination of the pleural fluid may give information about the local inflammatory reaction. Adenosine deaminase activity (ADA, a marker of TB pleurisy) was also tested. Tumour necrosis factor, IL-1 and ADA levels were measured in the pleural fluid and serum of 97 patients; 33 with tuberculous effusion, 33 with malignant effusion, and 31 patients with benign non-tuberculous effusion. Pleural fluid TNF and ADA levels were higher in tuberculous (TB) patients than in patients with benign disorders or cancer (P < 0.01). Serum TNF levels were also higher in TB patients than other benign (P < 0.01) or malignant (P < 0.05) effusions. There was a positive correlation between serum and pleural fluid values (r = 0.998-0.999, P < 0.001) although pleural fluid concentration was higher (P < 0.001), possibly suggesting local production in the pleural cavity. Pleural fluid IL-1 levels were not raised in any patient group but there was a positive correlation between TNF and IL-1. In addition, a positive correlation was found between TNF and ADA levels, probably indicating some common production mechanism. Furthermore, ADA sensitivity in the diagnosis of tuberculous effusion was augmented by the combined use of TNF and ADA. The use of both these markers may prove useful in the differential diagnosis of TBC pleurisy. PMID- 8730329 TI - Prevalence of asthma in young cross-country skiers in central Scandinavia: differences between Norway and Sweden. AB - An increased prevalence of asthma in cross-country skiers has been reported previously. This study was conducted in order to evaluate the prevalence of asthma among young, elite cross-country skiers in Trondelag, Norway and Jamtland, Sweden. These two regions in central Scandinavia have different climates during the winter season, with a coastal type in Trondelag while Jamtland is characterized by a colder, drier, inland type of climate. The study population consisted of 171 skiers (118 from Norway, 53 from Sweden). The investigation included an asthma questionnaire, spirometry and methacholine provocation testing. The prevalence of self-reported asthma-related symptoms was 46% in Norway and 51% in Sweden. The prevalence of bronchial hyper-responsiveness (BHR) was significantly different between the two regions. In Norway, 14% of the skiers were hyper-responsive to methacholine compared to 43% in Sweden (P < 0.001). Moreover, the estimated prevalence of clinically diagnosed asthma was 12% in Norway and 42% in Sweden (P < 0.001). The self-reported frequency of respiratory allergy was higher in Sweden (32%) than in Norway (11%). However, on an individual basis, self-reported allergy did not predict occurrence of asthma or BHR. In conclusion, there is a high prevalence of BHR and asthma among young, elite cross-country skiers in central Scandinavia, especially in Sweden. Cross country skiing may be a risk factor for the development of asthma. Longitudinal studies of cross-country skiers, and studies to acquire further knowledge of the mechanisms involved in the development of asthma are indicated. PMID- 8730330 TI - The effect of a time delay on the measurement of capillary blood gases. AB - Patients using domiciliary nasal ventilation, or long-term oxygen, require regular assessment which could be carried out in the home. Blood gas analysis may be regarded as an essential part of the assessment. The present study investigated the effect of a 1-h time delay on the measurement of capillary blood gases. Four samples of arterialized earlobe blood were collected from 15 outpatients. One sample was analysed immediately, the other three were stored on crushed ice and analysed at intervals of 30, 45, and 60 min post-collection. In order to examine any range effect, a wide range of PaO2 values were examined. The delay, at all levels, resulted in minor changes in the measurement of PaO2, which would be unlikely to alter clinical management. The technique might be used for the reliable assessment of patients in the home. PMID- 8730331 TI - Multiple pulmonary chondrohamartomas in trachea, bronchi and lung parenchyma. Review of the literature. AB - Multiple pulmonary chondromatous hamartomas (MPCH) are rare, and MPCH with tracheal involvement have not been, to the best of our knowledge, reported before. Furthermore, there is no agreement in the literature about the origin of these tumours. We report a case of MPCH involving trachea, bronchi and lung parenchyma, incidentally found at the autopsy of a 88-year-old woman who died of acute myocardial infarction, and we review the literature about MPCH. We conclude that there appear to be two different types of multiple pulmonary chondromatous hamartomas; those presented in young patients often linked to gastric leiomyoblastomas and catecholamine-producing paragangliomas (Carney syndrome) with high mortality, and those presented in elderly patients, which seems to be compatible with life. PMID- 8730332 TI - Traumatic lung cyst. PMID- 8730333 TI - Amiodarone pneumonitis: no safe dose. PMID- 8730334 TI - Death in notified cases of tuberculosis in Edinburgh: 1983-1992. PMID- 8730335 TI - Spontaneous closure of a large emphysematous bulla. PMID- 8730336 TI - Pulmonary lymphangioleiomyomatosis and tuberous sclerosis. PMID- 8730337 TI - Beta interferon and multiple sclerosis: why the fuss? PMID- 8730338 TI - Primary biliary cirrhosis. PMID- 8730339 TI - Response to trimethoprim/sulfamethoxazole in Wegener's granulomatosis depends on the phase of disease. AB - We prospectively studied trimethoprim/sulfamethoxazole (T/S) in inducing remission in 'initial phase' Wegener's granulomatosis (WG), and in sustaining remission in generalized WG, in 72 patients in various disease stages. Nineteen patients with initial phase WG received T/S (2 x 960 mg/day). Another 24 patients with generalized WG received the same dose of T/S (group A) and were compared with 21 patients receiving no further treatment after standard therapy (group B). Eight patients were given T/S plus low-dose prednisone (group C). Eleven of 19 patients (58%) with initial phase WG achieved complete or partial remission lasting a median 43 months (range 6-88 months). Of the remaining eight (42%), five showed local disease progression, and three developed generalized WG. In group A (T/S alone, generalized WG), 10/24 (42%) suffered a relapse after a median 13 months (range 4-58 months). In group B (generalized WG, no further treatment) 29% of patients relapsed after a median 22.5 months (range 18-26 months). All eight patients treated with T/S plus low-dose prednisone (group C) suffered serious relapse after 2-24 months. T/S induced long-term remission in > 50% of patients with initial phase WG; however, neither T/S alone nor T/S plus low-dose prednisone sustained remission in generalized WG. PMID- 8730340 TI - Neurotoxicity, anticoagulant activity and evidence of rhabdomyolysis in patients bitten by death adders (Acanthophis sp.) in southern Papua New Guinea. AB - Thirty-two patients with enzyme-immunoassay-proven death adder (Acanthophis sp.) bites were studied in Port Moresby, Papua New Guinea. Eighteen were envenomed; local signs were rare and none had incoagulable blood, but all except one had signs of neurotoxicity. Five (27.7%) envenomed patients required intubation and ventilation. One patient developed renal failure, previously undescribed following death adder bites. Laboratory investigations showed mild prolongation of prothrombin and partial thromboplastin times in some patients. In vitro studies showed that the venom contains anticoagulant activity, but does not cause fibrinogenolysis. In contrast to taipan envenoming, neurotoxicity did not progress after antivenom administration, and there was reversal of neurotoxicity, evident within 6 h, in three severely envenomed patients treated less than 12 h after the bite. One patient treated with antivenom and anticholinesterases had the most dramatic response to treatment; the optimum management of bites by this species may include prompt treatment with both antivenom and anticholinesterases in addition to effective first aid. PMID- 8730341 TI - Clinical spectrum, investigations and treatment of Budd-Chiari syndrome. AB - The clinical presentation, investigations, therapeutic modalities, prognosis and outcome of 44 patients with Budd-Chiari syndrome (BCS) were reviewed. There were 27 women and 17 men. Median age at presentation was 37 years (range 14-60). Possible aetiological factors were identified in 31 patients (70%). Myeloproliferative disorders were the commonest aetiology. Abdominal pain and swelling were the commonest presenting symptoms. Sixteen patients underwent a shunt operation (14 mesocaval, 2 mesoatrial). Ten patients had liver transplantation. Eleven patients had angioplasty/stent as their only treatment, and seven were treated medically or died before any treatment was instituted. Radiological intervention was comparable to mesocaval shunt in relieving patients' symptoms and in achieving good long-term results. Medical treatment and liver transplantation both yielded poor results. Radiological intervention in the form of balloon angioplasty or stent placement gives good results in a subgroup of BCS patients, and should be tried first to relieve the hepatic outflow obstruction. Mesocaval shunts provide good results in selected cases. Underlying haematological causes should be intensively investigated and promptly treated. PMID- 8730342 TI - Iron, ascorbate and copper status of Sowetan Blacks with calcific chronic pancreatitis. AB - Vitamin C can be used to overcome oxidative stress and ease pain in chronic pancreatitis. But its use is deprecated in conditions of tissue iron overload, because its bioactive form, ascorbate, can accelerate free-radical reactions that are driven by transition metals. We measured iron, ascorbate and copper in Sowetan Blacks (RSA) with chronic pancreatitis, obtaining serum/plasma from 14 consecutive patients and 15 controls. Compared with data from corresponding groups in Manchester, African samples had less ascorbate (p < 0.0001), but more caeruloplasmin (p < 0.0001). African and British controls had comparable iron and iron-binding capacity. Plasma from African patients had less ascorbate than that from African controls (p < 0.005) and in six samples, ferritin exceeded 300 micrograms/l (677 pmol/l). Low-molecular-mass iron or copper, capable of participating in free radical reactions, was not detected. British patients, had similar caeruloplasmin levels to African patients but higher ascorbate levels. There is no evidence of iron overload in our African samples. Outwardly healthy controls from Soweto have elevated levels of caeruloplasmin, possibly to compensate for dietary deficiency of ascorbate. Persistent oxidative stress is a unifying feature of chronic pancreatitis, but its degree is higher in African than British patients. Supplements of vitamin C should be safe in Blacks of southern Africa. PMID- 8730344 TI - Calcium channel blockers versus ACE inhibitors as antihypertensives in polycystic kidney disease. AB - The effects of calcium channel blockers (CCBs) and angiotensin converting enzyme (ACE) inhibitors on blood pressure and the progression of renal dysfunction were compared in hypertensive patients with polycystic kidney disease (PKD). Twenty six patients with PKD and hypertension who had been treated with other antihypertensive agents, such as diuretics, beta-blockers, or alpha-methyldopa, were followed up for two years, during which their blood pressure and renal function were monitored. Patients were divided into two groups classified according to the type of antihypertensive agents given. Group 1 (n = 14) received CCBs, while group 2 (n = 12) received ACE inhibitors. No significant differences were found in their blood pressure control and serum creatinine levels throughout the study. The creatinine clearances were decreased in both groups. However, the decreases in creatinine clearance were smaller (p < 0.05) in the group treated with CCBs. In addition, two patients in group 2 showed rapid increases in serum creatinine. Our data suggest that CCBs reduced blood pressure effectively and preserved renal function in PKD patients at least as well as ACE inhibitors. PMID- 8730343 TI - Cerebrotendinous xanthomatosis: a family study of sterol 27-hydroxylase mutations and pharmacotherapy. AB - We examined the phenotypic characteristics, molecular genetics and optimal pharmacological treatment of cerebrotendinous xanthomatosis (CTX) in an English family with combined hyperlipidaemia. The proband presented in adulthood with classical clinical characteristics of CTX, a greater than tenfold elevation in plasma cholestanol and combined hyperlipidaemia. His brother also had typical features of CTX without the presence of dyslipidaemia. Genotyping revealed that the two brothers were compound heterozygotes for a novel missense mutation in exon 2 (R94Q) and for a recently described nonsense mutation in exon 5, of the sterol 27-hydroxylase gene (CYP27). Analysis of all available family members revealed that hyperlipidaemia did not co-segregate with the presence of a CYP27 mutant allele. Trial of therapy showed that the lowest plasma sterol and triglyceride concentrations and cholestanol:cholesterol ratio were achieved with the combination of chenodeoxycholic acid (CDCA) 750 mg/day, a primary bile acid, and simvastatin 40 mg/day, an inhibitor of 3-hydroxy-3-methyl-glutaryl coenzyme A reductase. CDCA alone and simvastatin alone significantly lowered plasma cholestanol concentration, but the decrease was greater with the former. After 1 year there was significant improvement in both cognitive and motor function with regression of tendon xanthomata on computerized tomography. We conclude that CTX in this English pedigree is probably due to compound mutant alleles in CYP27, that combined hyperlipidaemia in this family is unrelated to CTX, and that this complicated condition responds optimally to the combination of CDCA and simvastatin. PMID- 8730345 TI - Bacterial meningitis in patients with nasopharyngeal carcinoma. AB - Bacterial meningitis was found in 12 patients with nasopharyngeal carcinoma, accounting for 0.65% of the 1850 patients with the tumour diagnosed between 1981 and 1994 in our hospital. In 11 patients, the time-lag between diagnosis of cancer and the appearance of infection ranged from 9 months to 11 years (mean 57 months) whereas in one patient it was only 5 days. Three patients developed mixed bacterial meningitis. Cerebrospinal fluid culture for bacteria was positive in six patients. Three patients (25%) were bacteraemic. Gram-negative bacilli, especially Pseudomonas aeruginosa, were the most common pathogens. Age, sex and histopathology were not risk factors for infection. Conditions predisposing to meningitis included intracranial invasion of the tumor, neutropenia, otitis media, and neurosurgical procedures. All but two patients had intracranial tumour invasion and erosion of the base of the skull. Local spread of micro-organism to the meninges was more important than haematogenous spread. The overall mortality in our patients was 66.7%, much higher than in patients without cancer. PMID- 8730346 TI - Economic and social impact of migraine. AB - We sent an anonymous self-administered questionnaire to 4200 employees of a Trust hospital, 1903 of whom returned it. We identified 158 migraine sufferers according to International Headache Society (IHS) criteria. These sufferers estimated 2.0 days/year absence from work, and an equivalent of 5.5 days/year lost by reduced effectiveness at work, caused by their migraine at an estimated financial cost of over 50,000 pounds to the Trust. An additional 220 individuals who had received a diagnosis of migraine from a doctor but fulfilled only 3/4 IHS criteria lost the equivalent of 6.7 days off work at a further cost of 63,000 pounds). Few patients had consulted their general practitioner about their migraines in the last 3 months. Most (78%) were using only over-the-counter medication. Migraine patients should be encouraged to seek medical attention. PMID- 8730347 TI - The NHS and the new scientism: solution or delusion? PMID- 8730348 TI - Priority for coronary artery surgery. PMID- 8730350 TI - Direct activation and defibrillation of cardiac tissue. AB - It is shown that inhomogeneity due to gap junctional resistance allows an excitable cable to be directly activated or defibrillated by the application of a large amplitude point stimulus of short duration. Multiple-scale analysis is used to derive a model equation describing the response of the excitable medium to stimuli, and this equation is analyzed to give quantitative estimates of the direct stimulus threshold and the defibrillation threshold. These estimates are shown to be in good agreement with experimental findings. PMID- 8730349 TI - A model of growth and form based on adhesion molecules. AB - A model is given for the generation of pattern and form in living systems, based on the assumption of two types of a single adhesion molecule that form homotypic cell-cell contacts. The time dependence of the model is different from that of the Turing models, instead viewing the change in time as being driven by the change in time of the total area A, along with the sequential gene activation which is called into play at discrete times as the organism grows and changes shape. Two equations are derived on the assumption of an energy minimum equilibrium being achieved at all times on a scale that is short compared with the rate of change over time of the total area. The two equations derived from the simple assumptions of the model are the (nonlinear) Helmholtz equation and the Laplace equation. It is argued that a suitable "morphogen" should attempt to satisfy certain rather specific conditions. PMID- 8730351 TI - Vestibular neurectomy: a histological and clinical study of results. AB - Posterior fossa vestibular neurectomy has become one of the most effective surgical procedures for control of vertigo symptoms in Meniere's disease. A small group of patients continue to have vertigo and demonstrable vestibular function by electronystagmography (ENG) post-operatively. Another group of patients may have no vertigo, despite residual vestibular function on caloric testing. In an effort to correlate histological findings with clinical outcome, nerve biopsies were taken from seven patients undergoing vestibular neurectomy. Fibre counts of nerve biopsies were compared with clinical, audiometric and ENG results. Results of this study demonstrate that complete reduction of vestibular response (on caloric testing) and clinical cure of vertigo can be obtained despite a variable number of nerve fibres at vestibular neurectomy. PMID- 8730352 TI - Disability after labyrinthectomy. AB - A labyrinthectomy is known to relieve vertigo successfully in the majority of patients who suffer from Meniere's disease and have non-serviceable hearing in the affected ear. It is assumed that the procedure reduces disability, helps the patient to return to work and improves the quality of life. Eighteen patients who underwent a transmastoid drill-out labyrinthectomy between 1980 and 1990 were interviewed and an attempt was made to evaluate the success of the operation in accordance with the guidelines set out by the AAO-HNS 1985. In the present series it was noted that although vertigo was relieved in 89 per cent of patients after labyrinthectomy, only 50 per cent of them returned to work. In this study, the age and occupation of the patient at the time of surgery and the relief of vertigo did not accurately predict whether or not a patient returned to work. PMID- 8730353 TI - Temporal bones for dissection. A diminishing asset? AB - In this prospective study, the author studied four local ENT departments to discover how many temporal bones were being harvested. The results show that only eight per cent of routine post mortems have temporal bones removed. PMID- 8730354 TI - Obstruction of the nasal valve. AB - Obstruction of the nasal valve is an important cause of chronic nasal obstruction in adults. In a series of 500 patients, obstruction at the level of the nasal valve was diagnosed in 65 of them (13 per cent). The obstruction was unilateral in 57 patients (88 per cent). Forty-seven patients (72 per cent) had history of previous nasal surgery of accidental trauma. Causes of obstruction of the nasal valve included high septal deviations, a weak or deformed upper lateral cartilage, adhesions, and alar collapse. All patients underwent corrective nasal surgery and the surgical procedures were tailored according to the existing pathology. Post-operatively, the mean nasal patency score increased from 2.9 to 8.6, the mean nasal airflow increased from 579.5 to 727 cm/sec (at 150 Pa), and the mean nasal resistance decreased from 0.31 to 0.23 Pa/cm3 sec-1. PMID- 8730355 TI - Levels of substance P and IgE decapeptide in nasal polyp fluid and matching sera: a preliminary study. AB - Sixteen unselected patients with nasal polyps had the levels of substance P and IgE decapeptide measured by ELISA in the oedema fluids and their matched sera. All 16 samples had low levels of substance P in their sera and had high level of substance P in eight samples of nasal polyp oedema. There was a considerable variation in the values of IgE decapeptide found in the sera but 14 polyp oedema fluids had high levels of IgE decapeptide. This study supports the idea that there is a linkage between the cellular and neurovascular responses. High levels of IgE decapeptide suggest that mast cell reactions occur in the majority of cases and that IgE may be implicated in the process of mast cell degranulation. PMID- 8730356 TI - Modifications of the Caldwell-Luc procedure for the prevention of post-operative sensitivity disorders. AB - The effectiveness of some modifications of the Caldwell-Luc standard procedure in the prevention of post-operative mid-facial sensitivity disorders was investigated in a prospective comparative study at the ENT Department, Faculty of Medicine, University of Athens, Greece, from 1986-1992. Sixty-nine maxillary sinuses were operated on in 61 patients suffering from benign pathological conditions. The modifications concerned the type of mucoperiosteal incision and the site and size of the created anterior antrostomy. Only 8.7 per cent of the sinuses operated on by the modified technique presented with post-operative disorders whereas among those operated on by the classical technique, the ratio climbed to 33 per cent. In view of the findings of this study, the modifications applied can be valuable in treating non-extensive maxillary sinus disease with minimal post-operative sensitivity disorders. PMID- 8730357 TI - Laser-assisted uvulopalatoplasty: an assessment of a technique. AB - The Kamami technique of laser-assisted uvulopalatoplasty has recently been introduced into British practice as an office-based multistage procedure for the treatment of snoring and, in certain cases, for obstructive sleep apnoea. We have treated 50 patients with simple snoring, with a minimum follow-up of six months. Six-month telephone interview follow-up has assessed partner and patient satisfaction, as well as willingness to undergo the procedure again. We have found the procedure easy to perform with no serious intra-operative complications. Thirty-four (68 per cent) of patients' partners have been definitely satisfied with the results of treatment, with 11 (22 per cent) definitely unsatisfied. However, only 25 of these 34 satisfied patients (76 per cent) would be prepared to undergo the treatment again with post-operative pain and discomfort cited as the reason in those who would not. The failure rate doubled between one and six months post-operatively. There have been no serious complications but a 40 per cent minor side-effect rate is noted. These results are discussed in the context of previously reported results, with consideration of future strategies to improve outcome. PMID- 8730358 TI - DNA indices of primary and recurrent squamous cell carcinomas of the tongue and tonsil using image cytometry. AB - Ploidy status of squamous cell carcinomas of the head and neck (SCCHN) from primary and recurrent tonsillar and tongue lesions has not been compared using image cytometry. We have measured and compared the DNA indices in 41 cases. There were 29 tongue SCCHN, 20/29 were primary and 9/29 were recurrent. Mean DNA index (DI) was 1.19 (range 0.70-1.81) and 1.28 (range 0.79-1.94) respectively. There were 12 tonsillar cases, 10/12 primary and two out of 12 recurrent. Mean DI was 0.84 (range 0.57-1.09) and 1.00 (range 0.98-1.02) respectively. Mean DNA indices of both primary carcinomas were lower than the mean DNA indices of the recurrent carcinomas. This difference between the two groups may be a reflection of their tumour biology. However, since our study is small no definite conclusions can be made at this stage. We aim in the future to evaluate the prognostic role of DNA indices of patients with paired primary and recurrent SCCHN. This may be of clinical value and improve the treatment modalities available to this group. PMID- 8730359 TI - Candida'the cancer of silastic'. AB - Candida species have the propensity to colonize and erode into the surface of silastic causing a loss of function. Colonization of silastic tracheo-oesophageal speaking-valves is an important cause of valve failure post-laryngectomy: the exact nature of the colonization is unknown and light microscopic studies have not been reported previously. Microbiological examination of scrapings taken from the colonized oesophageal surface of failed speaking valves was arranged before sectioning for light microscopy. Conventional ultrathin sectioning of silastic was difficult, therefore 15 micrometre sections were cut after cooling the material to -30 degrees C. These sections were then stained with toluidine blue; Candida species taking up the stain. Microbiological examination of the colonies confirmed the predominance of Candida species in the biofilm. Light microscopy of the 15 micrometer sections revealed that the Candida colonies had invaded the silastic. This work demonstrates that the colonization of the silastic valve prosthesis in the oesophageal area is due predominantly to Candida species which actively invade the structure of the silastic. This is an important cause of speaking-valve failure post-laryngectomy. PMID- 8730360 TI - Predictive value of flow cytometric analysis in DNA contents in patients with locally advanced head and neck carcinoma. AB - A retrospective study was performed on 61 eligible patients with stage III and IV (AJC/UICC Staging System) squamous carcinomas of the head and neck region who were treated with definitive radiotherapy with, or without, surgery. DNA contents were measured by flow cytometric analysis of archival paraffin blocks and were correlated with clinicopathological findings, tumour response and patient survival. Comparison of variables including treatment modality was performed for identification of significant prognostic factors. There were 28 diploid, 27 aneuploid tumours and the remaining six were questionable. All patients were followed-up for at least two years or until death. Aneuploid tumours had a significantly higher S-phase fraction (percentage S-phase) (p < 0.001). Neither ploidy nor percentage S-phase were found to have predictive value in tumour response or patient survival within the power of a sample size of 61. Twenty of the 27 (74 per cent) aneuploid tumours had a complete response (CR) whereas 19 out of 28 (68 per cent) diploid tumours achieved CR. Five-year survival by the Kaplan-Meier method was 33 per cent for both aneuploid and diploid tumours. However, nodal stage (N stage) was found to have significant predictive value in both tumour response and patient survival. The complete response for stage N0 patients was 96 per cent, N1 patients 61 per cent, N2 patients 60 per cent and 43 per cent for N3 patients (p < 0.002). Similarly, the five year survival for the N0 and N3 groups of patients was 53 per cent and 29 per cent respectively (p < 0.05). PMID- 8730361 TI - Mucoepidermoid carcinoma of the middle ear-a case report. AB - We report a case of mucoepidermoid carcinoma of the middle ear, a site which does not appear to have been previously described. A discussion of four possible theories of pathogenesis are presented. PMID- 8730362 TI - Hearing loss in multiple sclerosis: localization of the auditory pathway lesion according to electrocochleographic findings. AB - Multiple sclerosis is known to affect the myelin of the auditory pathway resulting in acute hearing loss. Two cases of sudden deafness due to multiple sclerosis have been evaluated by conventional audiometry, brainstem auditory evoked response audiometry and transtympanic electrocochleography. The abnormalities of the compound action potential in both patients (enhanced latency, abnormal adaptation using fast stimulus rate) and the normal receptor potentials (cochlear microphonic, summating potential), as well as the absence of brainstem responses suggest a disturbance of synchronization at the level of the first auditory neurone. The electrocochleography provides valuable information for the topodiagnosis of this and other neural hearing losses, especially in the absence of reliable brainstem responses. PMID- 8730363 TI - Air travel induced meningitis following vestibular schwannoma (acoustic neuroma) surgery. AB - Two cases are presented in which patients flying home shortly after translabyrinthine removal of a vestibular schwannoma (acoustic neuroma) developed acute bacterial meningitis on landing. This complication has not been described before. The incidence and management of CSF leaks after vestibular schwannoma surgery is also discussed. PMID- 8730364 TI - Blindness due to electrocoagulation following functional endoscopic sinus surgery. AB - A case is presented with irreversible blindness in one eye and restriction of the infratemporal visual field in the other eye, due to electrocoagulation performed for delayed bleeding after functional endoscopic sinus surgery (FESS). Although blindness due to rhinosurgery is a well-known, but rare complication, a review of the recent literature did not reveal a similar incident. The use of electrocoagulation especially under conditions of poor visualization, that may be the case in post-operative bleeding following (endoscopic) sinus surgery, is strongly discouraged. PMID- 8730365 TI - Proptosis following nasal tamponade. AB - A fracture of the lamina papyracea as a result of nasal packing, though plausible is undocumented. A case is presented, with a cautionary note to avoid the use of excessive pressure in packs and balloons when attempting to control epistaxis in elderly patients. PMID- 8730366 TI - Surgical bone wax causing epistaxis. AB - A case of recurrent epistaxis caused by bone wax in the nasal cavity is reported. This is the first reported case of a nasal complication due to surgical bone wax. PMID- 8730367 TI - Nasal lipomas presenting as part of a syndromic diagnosis. AB - An important part of the initial assessment of children presenting with congenital nasal masses is to exclude an intracranial extension using either magnetic resonance imaging (MRI) or computed tomography (CT) imaging. We present three patients with such lesions in which unusual radiological findings were noted as part of the investigations. PMID- 8730368 TI - Bilateral antrochonanal polyps in a child: a case report. AB - In this paper we present the first ever reported case of simultaneously occurring bilateral antrochoanal polyps in a fit 12-year-old child. The antrochoanal polyps (ACP) were removed using functional endoscopic sinus surgery (FESS) which achieved complete cure. Histological analysis of the polyps showed them to be of benign inflammatory origin. PMID- 8730369 TI - Nasopharyngeal lipoma--a rare clinico-pathological entity. AB - A lipoma situated within the posterior wall of the nasopharynx is reported. Only three lipomas of the nasopharynx in adults have previously been reported. The value of CT scan and cytology in making the preoperative diagnosis of a lipoma at this location is discussed. PMID- 8730370 TI - Intraductal papilloma as a cause of a parotid duct cyst. A case report. AB - A case is presented of a rare intraductal papilloma being found simultaneously with a parotid duct cyst, which is a relatively common parotid disorder. The papilloma most probably played a significant role in the obstruction of the duct and thus caused the ductal cyst. Intraductal papillomas have been mostly described in minor salivary glands, but are rare in the parotid gland. PMID- 8730371 TI - An unusual foreign body migrating from pharynx to mediastinum. AB - A case of a swallowed foreign body migrating from the pharynx into the soft tissues of the neck is presented, whose management was complicated by its undetected migration to the superior mediastinum occurring between radiological localization in the neck three days prior to exploration and its attempted removal. A median sternotomy was then required to retrieve the object. Radiological guidance with regard to the localization of a foreign body in the neck can only be relied on when performed immediately prior to surgical removal. PMID- 8730372 TI - Bilateral phrenic nerve injury after neck dissection: an uncommon cause of respiratory failure. AB - The case of a patient with carcinoma larynx who developed diaphragmatic paralysis and post-operative respiratory failure due to bilateral phrenic nerve injury is reported. The use of portable ultrasonography for an early diagnosis of diaphragmatic paralysis is discussed. PMID- 8730373 TI - Pathology in focus. Spindle-cell carcinoma of the nasal septum. AB - Spindle-cell carcinoma of the nasal septum is an extremely rare malignancy. Such a case along with the salient features of this variant of squamous cell carcinoma and a review of literature on carcinoma of the nasal septum is depicted. PMID- 8730374 TI - Bilateral hairy polyp of the oropharynx. AB - Hairy polyp of the nasopharynx is an unusual but well-recognized entity, generally presenting as a single mass at birth or in the first year of life. We describe the clinico-pathological features of a previously unreported bilateral hairy polyp in an adult and briefly discuss the pathogenesis of this condition. PMID- 8730375 TI - Tonsillar metastasis from adenocarcinoma of the stomach. AB - Metastatic carcinoma of the tonsil is rare with only some 100 cases having been reported in the literature, of which nine have resulted from stomach carcinoma. Tonsillar metastases rarely become apparent before the diagnosis of the primary neoplasm. We describe here the clinical and histopathological findings of a case of unilateral palatine tonsil metastasis as the first sign of a signet-ring cell carcinoma of the stomach. This has not been reported previously. PMID- 8730376 TI - Schwannoma of the larynx: a case report. AB - A case of schwannoma of the larynx is presented. Clinical findings are presented together with magnetic resonance images. The literature is reviewed. The surgical technique is discussed and the histology is described. PMID- 8730377 TI - Injuries related to car crime: the joy-riding epidemic. AB - 'Joy-riding' is the term used, somewhat inappropriately, for the offence of taking a vehicle without the owner's consent. In certain areas, mainly deprived inner-city estates, there has been an increase in this crime. The aim of this study was to investigate its impact on the workload of an inner-city teaching hospital's busy accident and orthopaedic departments. In this prospective study, all patients admitted to hospital as a result of road-traffic accidents (RTAs) were identified during a 9 month period. A total of 1576 patients were admitted to the trauma unit. One hundred and fifty-two admissions were as a result of RTA and 20 (13 per cent) of these patients had injuries as a result of car crime. Of this group, eight were severely injured (ISS > 16) and six of these were innocent bystanders. Three patients (one joy-rider and two innocent bystanders) died as a result of car crime. The average length of hospital stay was 12 days (1-62 days) and the hospital in-patient costs were estimated to be at least 5200 pounds per patient. Injuries related to car crime results in a significant amount of work and financial cost to the National Health Service. PMID- 8730378 TI - Management of missile wounds of the maxillofacial region during the 20th century. AB - Rapid evacuation from the scene of injury, the Advanced Trauma Life Support (ATLS) protocol and advancements in medical and surgical practice afford high standards of care for maxillofacial gunshot wounds (GSWs). This series of nine case reports illustrates both the changes that have occurred and the similarities that remain in the management of these wounds since the First World War. It serves as a reminder of the problems that may be faced managing maxillofacial injuries both in a future conventional war and also increasingly in the civilian population. PMID- 8730379 TI - Does the dangerous dogs act protect against animal attacks: a prospective study of mammalian bites in the accident and emergency department. AB - This comparative prospective study of mammalian bites attending one urban Accident and Emergency department before the implementation of the Dangerous Dogs Act 1991 and again 2 years later, was to see the effect of the Act on the pattern of injury. The study comprised a simple questionnaire detailing the injury, the implicated species, and the treatment or referral if applicable. In both groups studied (before and after implementation of the Act) 134 consecutive bites were recorded, contributing 1.2 per cent and 1.23 per cent of total attendances during these respective periods. Dogs were found to bite most commonly: in the pre legislative group 73.9 per cent were due to dog bites and in the post-legislation group 73.1 per cent. In both groups studied, human bites occurred as the second most common mammalian bite; 17.9 per cent in the pre-legislation group and 12.7 per cent in the post-legislation group. Human bites were as common as those from the most implicated breed of dog. In general human bites were found to require more active treatment and specialist referral. The study demonstrates the vast majority of such injuries are treated within Emergency departments. This study also shows how dangerous breeds compare with others that bite, demonstrating that these breeds contribute to only a small proportion of these injuries. This comparative study clearly demonstrates little impact on rate of attendances for such injuries since the introduction of the 1991 Act. If legislation aims to reduce and prevent injury from animal bites, in its present form it does little to protect the public; this study suggests a wider control of the dog population may be required. PMID- 8730380 TI - Involuntary defaecation in acutely injured patients: a useful clinical sign. AB - The phenomenon of involuntary defaecation following acute, severe injuries has never been documented before, but has been noted by those who work in major trauma units. The exact cause of the phenomenon is unknown. We prospectively noted 14 patients who involuntarily defaecated in a 3-month period and analysed their clinical findings retrospectively. Involuntary defaecation occurred only in patients with severe penetrating or blunt injuries. In those with precordial stabs, involuntary defaecation was particularly predictive of cardiac injury, and in all injuries was associated with higher surgical intervention and mortality rates. The occurrence of involuntary defaecation in injured patients is a useful sign that nearly always indicates the need for urgent intervention. PMID- 8730381 TI - Incidence of five common fracture types in an institutional epileptic population. AB - We have reviewed 4521 patient-years and analysed 185,066 seizures to calculate the incidence of five common fractures in an epileptic population. Only 25.0 per cent of these fractures were known to have occurred during a seizure. When age and sex matched against a 'normal population', there was an increased incidence of femoral neck fractures (5.2 x), inter-trochanteric fractures (9.9 x), ankle fractures (9.9 x) and proximal humerus fractures (4.2 x). Surprisingly, there was no increased incidence of wrist fractures. Overall, there was a 4.3 x increased risk for all fractures and 3.2 x increased risk of fractures not related to fits. There was no evidence for the first fracture occurring at a younger age in this population. The possible causes of this greatly increased risk are discussed. PMID- 8730382 TI - The posterior Monteggia: a pathological lesion? AB - Ulnar fracture with posterior dislocation of the radial head is thought to be a relatively rare injury. We have recently treated six patients with this posterior Monteggia fracture dislocation. Five of these patients were on long-term corticosteroid therapy. The pathophysiology, treatment and outcome of this complex injury are discussed. PMID- 8730383 TI - Accidents with horses: what has changed in 20 years? AB - Horse riding is a dangerous pastime with more accidents occurring per hour than during motor-cycling. Since a prospective survey of horse-related injuries conducted at a major centre in 1971-1972, equestrian groups and the medical profession have encouraged improvements in training and protective riding wear. By conducting a similar study at the same centre 20 years later we hoped to assess the effects of these measures on the pattern of injuries resulting from contact with horses. Patient and injury details were recorded prospectively for all those presenting to the Accident Service at Oxford during the whole of 1991. Total admissions fell by 46 per cent on average. Most of the decrease was due to a near fivefold fall in those admitted with head injuries (P < 0.001). A reduction in the severity of such injuries was associated with an increased use of riding helmets. However, the most commonly injured group remained amateur young female riders suggesting the need for increasing awareness and training of this group. In seven cases, severe digital injuries were caused by the habit of entwining reins around the fingers. This practice should be discouraged. Up to 12 per cent of all injuries might have been prevented if adequate footwear had been worn. PMID- 8730384 TI - The injury impairment scale in pelvic and lower limb fractures sustained in road traffic accidents. AB - To assess the validity of the Injury Impairment Scale for pelvic and lower limb fractures sustained in road traffic accidents, the Expected Injury Impairment Scale score was correlated with the Observed Injury Impairment Scale score for patients who attended the Accident and Emergency department in Nottingham during 1992 and 1993. Seventy-four per cent of the patients had an Observed IIS score equivalent to their Expected IIS score. The validity of the observed IIS scores for mobility and pain were assessed by correlating them with: (a) Functional assessment and visual pain scale scoring systems. A significant difference was found between scores 0 and 1 for both mobility and pain and between scores 1 and 2 for mobility only. (b) The physical functioning and pain parts of the Short Form 36 (SF36) disability questionnaire. A significant difference was found between scores 0 and 1 for both mobility and pain and between scores 1 and 2 for mobility only. The level of impairment was compared with the level of disability for mobility using the functional assessment score and the physical functioning part of the SF36 and for pain using the visual pain scale score and the pain part of the SF36 and no statistically significant relation was found. This study does not validate the IIS for pelvic and lower limb fractures. However, minor modifications would make it a valid and useful method for predicting the outcome of these injuries. PMID- 8730385 TI - Re-evaluation of two simple prognostic scores of outcome after proximal femoral fractures. AB - The Belfast and Newcastle simple prognostic scores of outcome after treatment of proximal femoral fractures were recorded shortly after admission on 250 patients. All patients were then followed until discharge, with 87 of those who were the more physically and socially dependent before admission being transferred after surgery for rehabilitation in a geriatric unit. The hospital mortality rate, a hospital stay of over 6 weeks and two measures of rehabilitation outcome, i.e. a delay of over 3 weeks after the operation in regaining independent mobility and any functional decline on discharge compared with the preadmission situation, were recorded for each patient. The prediction of a higher hospital mortality rate and a delay in reaching independent mobility in more dependent patients with both scores reached statistical significance. Longer hospital stays in the more dependent patients did not occur in a situation where care facilities for dependent patients were readily available. The scores, in particular the Belfast score, can be used to compare the casemix of patients with proximal femoral fractures within and between units over time and can predict mortality and delay in achieving independent mobility. They may be helpful in the early selection of patients for transfer to a geriatric unit or orthogeriatric unit for rehabilitation. PMID- 8730386 TI - Blunt abdominal injuries. Diagnostic peritoneal lavage, ultrasonography and computed tomography scanning. AB - The diagnosis of blunt abdominal injuries is one of the most difficult problems in the management of trauma. There is now better understanding of the diagnostic facilities available. Guidelines regarding the use of diagnostic peritoneal lavage, ultrasonography, or computed tomography scanning should be available in the Accident and Emergency department. PMID- 8730387 TI - Excision of heterotopic ossification around the knee following brain injury. AB - We present the results of five patients who underwent resection of heterotopic ossification around the knee following traumatic brain injury. All the patients had improved function and no pain at a minimum of 1 year follow up, irrespective of their preoperative neurological status. Our philosophy for such surgery is outlined and, although the surgical goals may seem small, the outcome for any individual patient may be considerable. PMID- 8730388 TI - Review of emergency thoracotomy for chest injuries in patients attending a UK Accident and Emergency department. AB - Over a two and a half year period, 25 patients presenting to the Glasgow Royal Infirmary underwent emergency thoracotomy for suspected severe chest injuries. Eighteen (72 per cent) were performed in the Accident and Emergency (A&E) department and seven (28 per cent) in a fully equipped operating theatre after resuscitation. There were 23 men and 2 women. Twenty-three (92 per cent) had been stabbed, one (4 per cent) had been shot and one (4 per cent) had sustained a blunt injury in a road traffic accident. Eight (32 per cent) patients survived. All survivors had been stabbed and seven were well enough to undergo thoracotomy in theatre. Only one (5.6 per cent) of the patients operated upon in the A&E department survived to discharge, although three (16.8 per cent) survived the initial procedure. Three of four patients survived, in whom the diagnosis of cardiac tamponade was initially missed. Thirteen (76.5 per cent) of the 17 who did not survive had no vital signs on admission. Outcomes may be improved if appropriately trained hospital staff are immediately available and prehospital delays are minimized so that patients arrive sooner with signs of life still present. Ambulance paramedic interventions have little to offer these patients and may worsen the prognosis if they result in delayed transport to hospital. The emphasis placed on diagnosis and treatment of cardiac tamponade in Advanced Trauma Life Support programmes is appropriate and all staff involved in these cases should undergo this type of training. PMID- 8730389 TI - Rehabilitation and reintegration of multiply injured patients: an outcome study with special reference to multiple lower limb fractures. AB - In this study a group of 104 multiply injured patients (mean ISS = 34) was examined for physical status and social reintegration. Essentially it was noted that 76 per cent of all patients were able to return back to work, although this appears to be age dependent. Open fractures of the leg seem to be the dominating prognostic factor for reintegration resulting in a disability rate of between 20 and 50 per cent. Disability of more than 80 per cent was only due to major head injury. Apparently reintegration is linked to the pattern of injury and is less related to the overall injury severity. We conclude that trauma care is economically cost effective but also needs further standardization and subsequent outcome analysis. PMID- 8730390 TI - External fixation in unstable intertrochanteric fractures of femur. AB - Forty elderly patients aged 50-94 years old with unstable intertrochanteric fractures were treated by external fixation at The Central Institute of Orthopaedics, Safdarjang Hospital, New Delhi, India from June 1993 to January 1995. Fracture union was achieved by an average time of 14 weeks in all the patients. Postoperative varus was increased in nine and deep pin-track infection was found in three cases. External fixation minimizes surgical damage and blood loss, and has an added advantage of early mobilization. This technique is simple, safe and economical. It is effective wherever early elective surgery is not possible due to limited resources. It is the method of choice for high-risk geriatric patients. PMID- 8730391 TI - Endothelial stimulating angiogenic factor in early fracture healing. AB - The early vascular response is essential for the normal progress of fracture healing and fracture site blood flow has been shown to reach a peak in the first two weeks after injury. Angiogenesis is an important step in this response as new vessel formation is necessary to vascularize the fracture haematoma and the fracture gap. Changes in serum levels of a low molecular weight endothelial stimulating angiogenic factor (ESAF) have been previously reported in a group of four patients with tibial fractures. In this group, ESAF levels were measured on three occasions only and at different time intervals. We present a more detailed profile of serum ESAF level changes in the first 14 days after the fracture. PMID- 8730392 TI - Non-union of a fracture of the 8th rib. PMID- 8730393 TI - Removing malleolar screws with ease. PMID- 8730394 TI - Sleep, dreams, and sudden death: the case for sleep as an autonomic stress test for the heart. PMID- 8730395 TI - Insulin and hypertension: a causal relationship? PMID- 8730396 TI - Vascular aneurysms: a side-splitting affair. PMID- 8730397 TI - Thrombin or platelets as major point of impact in vascular reocclusion and restenosis; a controversy? PMID- 8730398 TI - Thrombin plays a pivotal role in vascular re-occlusion after PTCA and coronary thrombolysis. PMID- 8730399 TI - Evidence for a pivotal role of platelets in vascular reocclusion and restenosis. PMID- 8730400 TI - Hypotonic-induced stretch counteracts the efficacy of the class III antiarrhythmic agent E-4031 in guinea pig myocytes. AB - OBJECTIVES: The aim was to determine the effect and mechanisms by which myocyte stretch interacts with the prolongation of action potential duration (APD) by the class III antiarrhythmic agent E-4031. METHODS: Action potentials and whole-cell currents were measured in isolated guinea pig ventricular myocytes with a patch clamp procedure during perfusion of normotonic, normotonic with addition of E 4031, and hypotonic plus E-4031 solutions. RESULTS: Cell swelling leading to membrane stretch of myocytes in the whole-cell recording configuration occurred with hypotonic solution perfusion. APD, prolonged by E-4031, was reduced to less than control value with hypotonic-induced stretch. Evaluation of whole-cell currents after hypotonic-induced stretch revealed no significant changes in the L type Ca2+ current, inward rectifier K+ current or the rapid component of the delayed rectifier K+ current. The slow component of the delayed rectifier K+ current (IKs) was upregulated and a stretch-induced CI- current was activated in hypotonic solutions. The hypotonic-induced modulation of these currents was not effected by protein kinase A or C inhibition. CONCLUSIONS: Hypotonic-induced stretch shortens APD and counteracts the effects of E-4031. This APD shortening is secondary to upregulation of IKs and activation of a stretch-induced Cl- current. PMID- 8730401 TI - The cellular electrophysiological effects of tedisamil in human atrial and ventricular fibers. AB - The cellular electrophysiological effects of 1 microM tedisamil (KC 8857) were studied in human atrial and ventricular fibers. Conventional microelectrode technique was applied to record the transmembrane action potentials at stimulation frequency of 100 per min and 37 degrees C. Tedisamil lengthened action potential duration (APD) more in atrial than in ventricular muscle fibers; prolongation of APD90 was 28.9 +/- 3.3% (n = 6; p < 0.05) for atrial and 13.3 +/- 5.2% (n = 6; P < 0.05) for ventricular tissue. The maximal rate of depolarization was depressed slightly, but significantly by 1 microM tedisamil only in ventricular fibers (12.9 +/- 6.5%, n = 6, P < 0.05). From these cellular electrophysiological data it is concluded that the bradycardic/antiischemic agent tedisamil possesses marked Class III properties not only in cardiac tissues of experimental animals but also those of man. PMID- 8730402 TI - Effect of a high omega-3 fatty acid diet on cardiac contractile performance in Oncorhynchus mykiss. AB - OBJECTIVE: Omega-3 fatty acids have been implicated in the amelioration of cardiovascular disease in humans. Since these fatty acids are found in salmonid fish and are known to be essential for all salmonids, this study was undertaken to determine the effect of a high dietary intake of omega-3 fatty acids on the function of trout myocardium. METHODS: Rainbow trout (Oncorhynchus mykiss) from a single stock population were divided into two groups and fed either a diet high in omega-3 fatty acids (i.e. 4.0%) or low in omega-3 fatty acids (i.e. 2.1%) for 3 months. Heart function was studied at the whole heart and isolated muscle level. RESULTS: In whole heart preparations, peak developed pressures in freely ejecting hearts from salmonids fed the high omega-3 fatty acid diet were significantly greater than the hearts from salmonids fed the low omega-3 fatty acid diet (21 +/- 1.5 vs. 11.5 +/- 0.9 mmHg respectively, P < 0.05). These data correlated with results from isolated muscle preparations of myocardium from fish fed high and low omega-3 fatty acid diets (4.12 +/- 0.32 vs. 3.08 +/- 0.28 mN/mm2 respectively, P < 0.05). The calcium uptake rate of heart homogenates from fish fed the high omega-3 diet was slower and sarcoplasmic reticulum Ca2+ ATPase activity was lower. The myofilament force-calcium relationship in myocardium from trout fed the low omega-3 diet was shifted leftward on the calcium axis to lower intracellular calcium concentrations (delta 0.4 pCa units) compared to mammalian myocardium. This resulted in greater activation at lower intracellular calcium concentrations. However, trouts fed diets high in omega-3 fatty acids had [Ca2+] required for half maximal activation more similar to what has been reported for mammalian myocardium (delta 0.1 pCa unit). Furthermore, the myofilaments of trout hearts appear to show less cooperativity (Hill coefficient approximately 1) than has been found in mammalian myocardium (Hill coefficient > or = 2). CONCLUSIONS: Our experimental results demonstrate for the first time that dietary omega-3 fatty acid content affects myocardial force of contraction by affecting calcium metabolism and myofilament calcium-activation. PMID- 8730403 TI - Effect of brief hypoxia on reperfusion arrhythmias and release of Ca2+ by rat heart homogenate blocked by ryanodine. AB - OBJECTIVES: We previously reported that a brief period of hypoxic perfusion (BHP) prior to ischemia in rat hearts improved functional recovery upon reperfusion with reduced Ca2+ overload. The present study was designed to determine whether the effect of BHP would be associated with a reduction in reperfusion arrhythmias and a preservation of function of the sarcoplasmic reticulum (SR). METHODS: Hearts were subjected to 40 min of global ischemia and 30 min of reperfusion after a 20 min period of oxygenated perfusion (oxygenated group: OG), or a 10 min period of oxygenation and 10 min of hypoxic perfusion (hypoxic group: HG). We evaluated the release of Ca2+ by SR blocked by ryanodine, the recovery of left ventricular function, and the reperfusion induced ventricular tachycardia/fibrillation (VT/VF). RESULTS: Functional recovery improved and the incidence and duration of reperfusion VT/VF were reduced in HG. In HG the uptake of Ca2+ in SR decreased during ischemia, but this decrease was less than that in OG. However, recovery of Ca2+ uptake after reperfusion did not differ between groups. The release of Ca2+ by SR blocked by ryanodine was inhibited in HG throughout the ischemia-reperfusion sequence. CONCLUSIONS: Observations suggest that the benefits of BHP on recovery of function and reperfusion arrhythmias were associated with a decrease in release of Ca2+ by SR blocked by ryanodine. PMID- 8730404 TI - Glyburide-reversible cardioprotective effects of calcium-independent phospholipase A2 inhibition in ischemic rat hearts. AB - OBJECTIVES: A myocardial calcium-independent PLA2 has been described that is activated during myocardial ischemia and this enzyme may modulate ATP-sensitive potassium channels (KATP). The aim of this study was to determine the effect of an inhibitor of this enzyme, a bromoenol lactone, in isolated globally ischemic rat hearts. METHODS: Isolated rat hearts were treated for 10 min with 0.3-6 microM bromoenol lactone and then subjected to 25 min ischemia and 30 min reperfusion. RESULTS: The bromoenol lactone significantly increased coronary flow in nonischemic myocardium, and slightly reduced cardiac function at 6 microM. During global ischemia, time to contracture was significantly increased from vehicle group values in the presence of the bromoenol lactone (EC50 = 1.2 microM). During reperfusion, a concentration-dependent increase in function and a reduction in LDH release were observed for the PLA2 inhibitor. The concentrations of the PLA2 inhibitor which were significantly cardioprotective, inhibited this enzyme in membrane fractions of rat myocardium (IC50 = 0.87 microM). The KATP blocker sodium 5-hydroxydecanoate (5-HD) inhibited the increase in time to contracture observed for the bromoenol lactone. During reperfusion, 5-HD abolished the protective effects of the bromoenol lactone on cardiac function and LDH release. Glyburide had similar effects on the cardioprotective activity of the bromoenol lactone, although it only partially abolished the LDH reducing effect of this agent. CONCLUSIONS: The bromoenol lactone protects ischemic myocardium at concentrations which also inhibit calcium-independent PLA2. This cardioprotection can be attenuated by blockers of KATP, suggesting a potential mechanism for modulation of myocardial KATP. PMID- 8730405 TI - Faster femoral artery blood velocity kinetics at the onset of exercise following short-term training. AB - OBJECTIVE: The hypothesis that the adaptation to endurance exercise training included a faster increase in blood flow at the onset of exercise was tested in 12 healthy young men who endurance-trained (ET) 2 h/day, for 10 days at 65% VO2 peak on a cycle ergometer, and in 11 non-training control (C) subjects. METHODS: Blood flow was estimated from changes in femoral artery mean blood velocity (MBV) by pulsed Doppler. Beat-by-beat changes in cardiac output (CO) and mean arterial pressure (MAP) were obtained by impedance cardiography and a Finapres finger cuff, respectively. MBV, MAP and CO were measured at rest and during 5 min of dynamic knee extension exercise. Both legs worked alternately with 2 s raising and lowering a weight (15% maximal voluntary contraction) followed by 2 s rest while the other leg raised and lowered the weight. RESULTS: In the ET group the time to 63% (T63%) of the approximately exponential increase in MBV following 10 days of training (8.6 +/- 1.2 s, mean +/- s.e.) was significantly faster than the Day 0 response (14.2 +/- 2.1 s, P < 0.05). The T63% of femoral artery vascular conductance (VCfa) was also faster following 10 days of ET (9.4 +/- 0.9 s) versus Day 0 (16.0 +/- 2.5 s) (0.05). There was no change in the T63% of both MBV and VCfa for the C group. The kinetics of CO were not significantly affected by ET, but the amplitude of CO in the adaptive phase, and at steady state, were significantly greater (P < 0.05) at Day 10 compared to Day 0 for the ET group with no change in the C group. CONCLUSIONS: These data supported the hypothesis that endurance training resulted in faster adaptation of blood flow to exercising muscle, and further showed that this response occurred early in the training program. PMID- 8730406 TI - Respective role of humoral factors and blood pressure in cardiac remodeling of DOCA hypertensive rats. AB - OBJECTIVES: Recent studies have shown that beside elevated arterial blood pressure, humoral factors such as angiotensin II, aldosterone, endothelin or bradykinin might play a role in the cardiac hypertrophy and fibrosis secondary to hypertension. In addition, it seems that perivascular fibrosis and interstitial fibrosis are controlled by independent mechanisms. Therefore, the goal of our study was to evaluate the respective role of the increased arterial pressure and of humoral factors on cardiac remodeling in an experimental hypertension model. METHODS: Uninephrectomized rats received DOCA, a high salt diet, and when hypertension was installed, they were treated for 6 weeks with either a long acting calcium antagonist, mibefradil (30 mg/kg day-1), an ACE inhibitor, enalapril (3 mg/kg day-1), or a mixed ETA-ETB endothelin receptor antagonist, bosentan (100 mg/kg day-1). A group of hypertensive rats was left untreated and a sham-operated group of normotensive rats was used for control. At the end of treatment, maximal coronary blood flow was measured in isolated perfused hearts. Cardiac hypertrophy and interstitial as well as perivascular fibrosis were evaluated by quantitative morphometry. RESULTS: DOCA-salt hypertensive rats exhibited a marked cardiac hypertrophy associated with a decrease of maximal coronary blood flow and interstitial and perivascular fibrosis. The calcium antagonist nearly normalized arterial pressure and suppressed all these changes. Enalapril had no effect on arterial pressure and perivascular fibrosis but decreased subendocardial fibrosis. Bosentan had a very small effect on arterial pressure but decreased cardiac hypertrophy and both perivascular and subendocardial fibrosis. CONCLUSIONS: We conclude that in DOCA salt hypertension, humoral factors such as endothelin may play a role beside high blood pressure in cardiac remodeling. In addition, the different components of this remodeling (decrease of vascular reserve, cardiac hypertrophy and cardiac fibrosis) are controlled independently. PMID- 8730407 TI - Plasma marinobufagenin-like and ouabain-like immunoreactivity during saline volume expansion in anesthetized dogs. AB - OBJECTIVES: This study investigated effects of acute plasma volume expansion on plasma levels and urinary output of two endogenous Na,K-ATPase inhibitors, marinobufagenin-like and ouabain-like immunoreactive substances. METHODS: Plasma volume was expanded for 3 h via intravenous saline infusion in three groups of anesthetized dogs--nontreated (n = 5); pretreated with rabbit antidigoxin (n = 5); and pretreated with rabbit antimouse (control) antibody (n = 4). RESULTS: Plasma marinobufagenin-like immunoreactivity increased to 11.87 +/- 3.16 nmol.l-1 (vs. 0.30 +/- 0.16 nmol.l-1) within 10 min of volume expansion, in parallel with a 15% increase in LVdP/dt, then decreased to 2.21 +/- 0.59 nmol.l-1, and in 90 min increased to 11.8 +/- 2.8 nmol.l-1, in parallel with the maximal natriuretic response. Plasma concentrations of ouabain-like immunoreactive material were increased after 90 min of saline infusion (0.019 +/- 0.004 nmol.l-1 vs. 0.139 +/- 0.056 nmol.l-1). Pretreatment of the animals with antidigoxin antibody blocked the positive inotropic and reduced natriuretic response to volume expansion, and decreased the urinary release of marinobufagenin-like, but not ouabain-like, material. CONCLUSIONS: These results show the presence of marinobufagenin-like immunoreactive substance in dog plasma and suggest that mammalian EDLF may have a bufodienolide nature. Endogenous marinobufagenin-like immunoreactive substance, which is likely to cross-react with antidigoxin antibody, is involved in the natriuretic and positive inotropic responses to plasma volume expansion. PMID- 8730408 TI - Specific mitochondrial DNA deletions in idiopathic dilated cardiomyopathy. AB - OBJECTIVE: Structural changes in human mitochondrial DNA (mtDNA) have been implicated in a number of clinical conditions with dysfunctions in oxidative phosphorylation called OX-PHOS diseases, some of which have cardiac involvement. The objective of this study was to assess the frequency and extent of specific mitochondrial DNA deletions in idiopathic dilated cardiomyopathy. METHODS: DNA extracted from tissue derived from the left ventricle of 41 patients with idiopathic dilated cardiomyopathy and 17 controls was amplified by polymerase chain reaction using specific primers to assess the incidence and proportion of 5 kb and 7.4-kb deletions in mitochondrial DNA. RESULTS: In reactions using primers to detect the 5-kb deletion, an amplified product of 593 bp was found in low abundance relative to undeleted mitochondrial DNA but with high frequency in a number of controls and patients. A second deletion of 7.4 kb in size was also frequently present in controls and patients. In contrast to previous reports, these deletions were found to be present in both controls and in cardiomyopathic patients, 18 years and younger, including several infants. The 7.4-kb deletion was prominently increased in both frequency and in its proportion relative to undeleted mitochondrial DNA in patients 40 years and older with idiopathic dilated cardiomyopathy. CONCLUSIONS: At variance with current literature our study reports a significant presence of both 5 and 7.4-kb deletions in the young and a higher frequency and quantity of the 7.4-kb deletion in the older cardiomyopathic patients in comparison with controls. The increased accumulation of the 7.4-kb deletion as both a function of aging and cardiomyopathy is suggestive that this specific mitochondrial DNA deletion arises more likely as an effect of heart dysfunction rather than as a primary cause of cardiomyopathy. PMID- 8730410 TI - Doxazosin blocks the angiotensin II-induced smooth muscle cell DNA synthesis in the media, but not in the neointima of the rat carotid artery after balloon injury. AB - OBJECTIVE: Infusion of angiotensin II (AngII) during the third and fourth week after balloon injury of the left common carotid artery of the rat induces smooth muscle cell (SMC) DNA synthesis. In this study we wanted to investigate whether alpha 1-adrenoreceptors are involved in AngII-induced SMC DNA synthesis in the neointima. METHODS: Adult male Wistar Kyoto rats were subcutaneously infused for 2 weeks with AngII and the alpha 1-adrenoreceptor antagonist doxazosin during the 3rd and the 4th week after balloon injury of the left common carotid artery. Control groups received AngII, 0.9% NaCl, AngII + 50% dimethylsulfoxide (DMSO, the solvent of doxazosin), doxazosin or 50% dimethylsulfoxide. Each rat received 5-bromo-2'-deoxyuridine in a separate osmotic minipump to label DNA-synthesizing SMC. Systolic blood pressures were measured in all groups. RESULTS: Angiotensin II caused an increase in systolic blood pressure, whereas addition of doxazosin did not affect the increase in SBP caused by AngII. In the media of the non injured carotid artery, AngII increased SMC DNA synthesis, as the BrdUrd labeling fraction increased from 0.2 +/- 0.1% (mean +/- s.e.m.) in the NaCl group towards 3.4 +/- 0.6% in the AngII group. Coinfusion with doxazosin reduced the AngII induced increase in BrdUrd labeling fraction from 3.2 +/- 0.8% in the AngII + DMSO group towards 0.6 +/- 0.2% in the AngII+doxazosin group. A similar effect of doxazosin was found in the media of the injured left carotid artery, in which coinfusion with doxazosin also reduced the BrdUrd labeling fraction from 2.6 +/- 0.8% in the AngII+DMSO group towards 0.3 +/- 0.1% in the AngII+doxazosin group. In the neointima of the injured left carotid artery, AngII increased the BrdUrd labeling fraction from 11.7 +/- 1.6% in the NaCl group towards 28.0 +/- 3.4% in the AngII group. Coinfusion with doxazosin did not influence the AngII-induced SMC DNA synthesis, since the BrdUrd labeling fraction in the neointima of the AngII+doxazosin group was 22.5 +/- 2.9%, whereas the neointimal BrdUrd labeling fraction in the AngII+DMSO group was 22.9 +/- 2.3%. Little effect was found on the medial cross-sectional area. The neointimal cross-sectional area was increased as a result of infusion of AngII (0.12 +/- 0.01 mm2 vs. 0.18 +/- 0.01 mm2), and coinfusion of doxazosin did not reduce the AngII-induced increase in neointimal cross-sectional area (0.18 +/- 0.03 mm2). CONCLUSIONS: These data suggest that alpha 1-adrenoreceptors are not involved in AngII-induced neointimal SMC DNA synthesis and cross-sectional area, but only play a role in the media of the carotid artery. PMID- 8730409 TI - Right and left ventricular geometry and myocyte contractile processes with dilated cardiomyopathy: myocyte growth and beta-adrenergic responsiveness. AB - OBJECTIVES: Comparison of the effects of supraventricular tachycardia-induced dilated cardiomyopathy on left and right ventricular isolated myocyte geometry and function. BACKGROUND: Chronic ventricular tachycardia and supraventricular tachycardia cause left ventricular dilation and dysfunction in humans. However, it is unknown whether supraventricular tachycardia-induced dilated cardiomyopathy is a homogenous process for both the left and right ventricles. METHODS: Dilated cardiomyopathy was induced by rapid atrial pacing (240 beats/min, 3 weeks) in 5 pigs. Five age- and weight-matched pigs served as controls. Ventricular mass was measured, myocyte dimensions were obtained, and isolated right and left ventricular myocyte contractile performance was evaluated at baseline and after beta-adrenergic receptor stimulation. RESULTS: With the development of dilated cardiomyopathy, there was no change in left ventricular mass. In contrast, right ventricular mass was increased, as was right ventricular myocyte cross-sectional area. In the control group, baseline right ventricular myocyte contractile function was increased compared to left ventricular myocytes. beta-adrenergic receptor stimulation increased myocyte contractile function in both left and right ventricular myocytes. With supraventricular tachycardia-induced cardiomyopathy, both left and right ventricular myocyte contractile function and beta-adrenergic responsiveness were reduced. CONCLUSIONS: This study demonstrated differences in left and right ventricular myocyte growth with supraventricular tachycardia-induced dilated cardiomyopathy and this differential growth response was associated with changes in contractile performance. Thus, in this model of cardiomyopathic disease, left and right ventricular growth and changes in contractile performance are not a homogenous process. PMID- 8730411 TI - Tenascin expression in nephrogenesis and in normal or pathologic glomerulus morphologic features and functional implications. PMID- 8730412 TI - Urea recycling: an aid to the excretion of potassium during antidiuresis. AB - Urea absorption in the inner medullary collecting duct provides a mechanism to elevate the concentration of urea in the papillary interstitial fluid and thereby permit the excretion of urea with as little water as possible. Urea reabsorption may have another important effect - to aid in the excretion of potassium (K). K excretion depends on two processes: first, factors such as aldosterone which cause the concentration of K in the luminal fluid of the cortical distal nephron to be high and, second, factors which augment the flow rate through those nephron segments. Since, the osmolality of the luminal fluid in the cortical collecting duct (CCD) and plasma are equal when antidiuretic hormone acts, the flow rate in the CCD is dependent on solute delivery. Urea is a major solute in the lumen of the CCD and thereby plays an important role in maintaining the CCD flow rate. Since urea and K are often found in the same foods, having urea help the excretion of K is potentially advantageous. If the excretion of urea was low, the flow rate in the terminal CCD would decline. In this circumstance, the luminal K concentration would have to rise in proportion to the fall in flow rate or there would be a diminished rate of excretion of K and, possibly, hyperkalemia. PMID- 8730413 TI - Study on proteinase-inhibiting capacity of plasma alpha 2-macroglobulin in idiopathic nephrotic syndrome. AB - We studied the relationship between plasma alpha 2-macroglobulin (alpha 2M) and depressed cell-mediated immunity in idiopathic nephrotic syndrome (INS). Plasma alpha 2M concentrations (mumol/l) were increased during relapses of INS; however, the proteinase inhibitory activity, measured using bacterial thermolysin, was significantly decreased when calculated per 1 mol of alpha 2M, implying a reduced proteinase-inhibiting capacity of alpha 2M. The decreased proteinase-inhibiting capacity of alpha 2M was associated with the inhibitory activity of plasma on normal lymphocyte blastogenesis. Purified alpha 2M, when complexed with chymotrypsin, intensively inhibited normal lymphocyte blastogenesis induced by concanavalin A, as compared with the free form of alpha 2M. From these results it is suggested that, although the amount of alpha 2M protein has increased in the plasma of INS patients during relapse, its binding capacity to proteinases has relatively decreased. The results of this study may provide speculation for both the well-known high plasma alpha 2M concentrations and the immunodepression, both of which have been observed in INS patients in the past few decades. PMID- 8730414 TI - Serum tumor necrosis factor in mesangial IgA glomerulonephritis with macroscopic hematuria in children. AB - Tumor necrosis factor (TNF)-alpha and interferon (INF)-gamma levels were measured in the sera obtained from 29 patients with IgA glomerulonephritis (IgA GN), 8 patients with minimal change nephrotic syndrome (MCNS) and 12 patients with upper respiratory tract infection (URI) without renal diseases in children. The serum TNF-alpha level of IgA GN was 123.0 +/- 175.4 pg/ml, MCNS was 4.9 +/- 4.0 pg/ml and URI was 10.5 +/- 4.5 pg/ml respectively. The serum TNF-alpha level of IgA GN was significantly higher than those of MCNS and URI. The serum TNF-alpha level of URI was on the high trend compared with that of MCNS, but was not statistically significant. Although the TNF-alpha level was related to mesangial cell proliferation in patients with IgA GN, it was unrelated to the grade of mesangial matrix expansion and magnitude of proteinuria. In 17 patients with IgA GN having macroscopic hematuria, the serum TNF-alpha level was 190.5 +/- 201.6 pg/ml, and in other IgA GN patients with microscopic hematuria it was 37.4 +/- 75.7 pg/ml. The serum TNF-alpha level of IgA GN with macroscopic hematuria was significantly higher than that with microscopic hematuria. In 6 patients with IgA GN with macroscopic hematuria, the serum TNF-alpha level was significantly decreased after macroscopic hematuria disappeared. The mean serum IFN-gamma level of IgA GN was 0.3 +/- 0.6 IU/ml, and MCNS was not detectable. Although the serum IFN-gamma level was related to mesangial cell proliferation in patients with IgA GN, it was unrelated to magnitude of proteinuria, the grade of mesangial matrix expansion and also the presence or absence of macroscopic hematuria. We suggest that macroscopic hematuria of IgA GN was closely related to the serum TNF-alpha level. PMID- 8730415 TI - Increased amounts of C3a and the terminal complement complex at high dialysis blood-flow: the relation with dialysis efficiency. AB - To estimate the influence of blood-flow on complement generation and the relation with dialysis efficacy (KT/V) 10 patients underwent cuprophan hemodialyses for 6 h using low (200 ml/min) or high (400 ml/min) blood-flow (n = 40). Dialysis with high blood-flow compared to low induced a more rapid drop in leukocyte count and a more pronounced leukocyte rebound. Net generation of C3a (microgram/min) was also larger at all 15 measuring points during high blood-flow dialysis and there was significantly larger total generation of C3a (after 3 h p < 0.05, after 6 h p < 0.05) as compared to low blood-flow. Reaching a KT/V of 1.2-1.4 with high blood flow induced a 40% larger total net generation of C3a and 21% more of TCC than with low blood-flow. Net generation of TCC (AU/min) was higher during the first and last parts of high blood-flow dialysis compared to low. In conclusion, increased blood-flow in order to increase dialysis efficiency (KT/V 1.2-1.4) results in larger net infusion of complement products with possible impact on immune response and complement-associated pathophysiological mechanisms. PMID- 8730416 TI - Intradialytic calcium balances with different calcium dialysate levels. Effects on cardiovascular stability and parathyroid function. AB - It has been shown that calcium carbonate (CaCO3) is an effective phosphate binder which is less toxic than Al(OH)3. However, given that its use with standard calcium dialysate (CaD) levels may lead to hypercalcemia, a decrease in CaD levels has been proposed. The aim of the present study was to elevate the acute clinical and biochemical consequences of a lowering of CaD in HD patients. Dialysate composition was otherwise the same. (1) Blood pressure levels (BP) during short hemodialysis were measured in a group of 12 patients who underwent alternate hemodialyses with dialysate calcium of 1.75 and 1.25 mmol/l. (2) Ca2+ and PTH kinetics during short hemodialysis were studied in a group of 6 patients who were sequentially treated with 1.75 and 1.25 mmol/l CaD. The results show: (1) that cardiovascular stability in chronic HD patients during short HD sessions with low CaD (LCaD) may be good; (2) that a single treatment with standard CaD (SCaD) produces positive calcium balances (JCa2+) with Ca2+ plasma increase and PTHi inhibition at the end of HD sessions; during HD with LCaD there were neutral mean JCa2+ and no changes in post-dialysis mean Ca2+ and PTHi plasma levels; furthermore 2 patients showed a small PTHi increase during HD with LCaD and neutral JCa2+ because of a high positive bicarbonate balance during HD. In conclusion, as with several aspects of dialysis treatment, dialysate calcium levels should also be individualized to avoid hypercalcemic crises or PTHi stimulation. PMID- 8730417 TI - Recombinant hepatitis B vaccine use in chronic hemodialysis patients. Long-term evaluation and cost-effectiveness analysis. AB - The prevalence of hepatitis B virus (HBV) infection in our unit was 45% (86/190); there were 77 (40.5%) and 9 (4.7%) patients with previous and persistent HBV infection, respectively. Recombinant hepatitis B vaccine was given to 118 chronic HD patients with a regimen of 3 double doses administered intramuscularly at 0, 1 and 2 months, obtaining a seroprotection rate of 67% (79/118), 57% (45/79) being high responders. At month 24, 78% (40/51) maintained protective levels of anti HBs, 45% (18/40) of them being high responders. There was a statistically significant difference between responder and non-responder patients with regard to nutritional parameters such as serum total proteins and mean levels of transferrinemia. The number of diabetic patients was significantly increased in the nonresponder group. Patients with persistent antibodies ('persistent responders') were younger and had a shorter duration of HD treatment compared to those responders who rapidly lost anti-HBs ('transient responders'). Serological positivity for antibodies against hepatitis B core antigen significantly facilitates the decrease of anti-HBs antibodies over time. We detected seven episodes of HBV infection among HD patients at our unit before the beginning of the vaccination program. On the contrary, there were no episodes of HBV infection among responder vaccinees during the 24-month follow-up period. After the initial cost of vaccination, a savings of US$ 3,272 per year was realized by the elimination of frequent serologic screening of vaccine responders. PMID- 8730418 TI - Normal fibrinolytic responses to 1-desamino-8-D-arginine vasopressin in patients with nephrogenic diabetes insipidus caused by mutations in the aquaporin 2 gene. AB - Three patients with autosomal-recessive nephrogenic diabetes insipidus (NDI), homozygous for mutations in the aquaporin 2 gene (AQP2), were tested for their fibrinolytic and hemodynamic responses to intravenous administration of 1 desamino-8-D-arginine vasopressin (DDAVP). They all showed an increase of tissue type plasminogen activator antigen, facial flushing, an increase of heart rate and a decrease of diastolic blood pressure. These results confirm the hypothesis that NDI patients with an AQP2 defect can be discriminated from NDI patients with a vasopressin type 2 receptor defect by their normal extrarenal responses to DDAVP. PMID- 8730419 TI - Experience with 100 combined pancreatic renal transplantations in a single center. AB - Compared to the still increasing number of kidney, liver and heart transplants performed worldwide, pancreas transplantation remains a rare occurrence. At our center a pancreas transplant program was began in late 1979. Since then a total of 113 pancreas transplants were performed in 106 patients, 100 of them also received a kidney from the same donor. The first group consisted of 5 patients with immediate duct occlusion (IDO). In the second group (n = 8) the pancreatic juice of the segmental graft was diverted into a Roux-Y loop of jejunum. Because of two fatal technique-associated complications, delayed duct occlusion was introduced and applied in 15 patients. Because of a prolonged hospitalization period due to local complications, the surgical technique was changed again. From 1987, 72 segmental pancreatic transplants with bladder drainage were performed and finally one whole organ with a duodenal segment was transplanted. Immunosuppression consisted of cyclosporine A, azathioprine and prednisolone from 1984 on. Rejection episodes were treated with a high-dose methylprednisolone on 3 consecutive days and steroid-resistant rejections with ATG. The overall patient survival at 6 years was 80%, renal allograft survival 72% and pancreas graft survival 63% for the entire group. In the delayed duct occlusion group, 1-year patient and kidney graft survival of 93% each and 79% for the pancreas was calculated. One-year survival in the most recent and largest group with bladder drainage was 89% for patients, 86% for the kidney and 75% for the pancreas. Excellent metabolic control was achieved in the majority of patients with mean C peptide levels and HbA1C levels at 6 months of 1.46 pmol/ml and 5.6%, respectively. Successful pancreas transplants with normalization of carbohydrate metabolism seem to have a beneficial effect on secondary complications of diabetes, contributing to the high degree of rehabilitation of these patients. PMID- 8730420 TI - The determination of plasma transferrin receptor as good index of erythropoietic activity in renal anemia and after renal transplantation. AB - Both the plasma determinations of erythropoietic (EPO) and transferrin receptor (TfR) would provide a good characterization of anemia especially when mixed erythron disorders underlie, such as in renal failure. Immunologic assays of EPO and TfR, as well as standard hematologic determinations (hematocrit, reticulocyte count, serum iron, transferrin, ferritin) were performed in patients with chronic renal failure (CRF), in regular dialysis treatment (RDT) and in transplanted (TX) patients. In nonanemic TX patients both EPO and TfR ranged normally, whereas in anemic TX ones (Hct < 40%) both values were increased suggesting the physiologic response both of the kidney and of the erythron to decreased red cell mass. In transitory posttransplant erythrocytosis the increased values of TfR, with normal EPO values, would hypothesize a defective feedback to EPO release. Both EPO and TfR values were found increased in TX patients with adult polycystic kidney disease with persistent erythrocytosis (Hct > 50%), thus confirming previous observations. In CRF and RDT patients, all anemic, both EPO and TfR were normal, even though significantly low with respect to the degree of anemia. In RDT seriously anemic patients, the administration of recombinant human EPO induced different patterns of bone marrow response. We conclude that the determination of TfR would provide further information on renal anemia since the receptor increase mostly preceded the rise of Hct, evidencing those patients who will not have an effective bone marrow response to the therapy. PMID- 8730421 TI - Validity in nulliparas of increased beta-human chorionic gonadotrophin at mid term for predicting pregnancy-induced hypertension complicated with proteinuria and intrauterine growth retardation. AB - The objective of the present study was to investigate whether increased beta human chorionic gonadotrophin (beta HCG) plasma concentrations in an unselected population of nulliparas could predict the occurrence of complicated pregnancy induced hypertension (PIH). The design was that of a prospective population study. It was conducted at the obstetric departments of Amiens University Hospital and Creil General Hospital on 434 consecutive nulliparas with singleton pregnancies after natural fertilization who accepted the systematic offer of trisomy 21 screening but for whom this disorder was finally estimated. Measurement of plasma concentration of beta HCG (ELISA method) was carried out between 14 and 20 weeks (mean: 17 weeks) of amenorrhea, and measurement of blood pressure and proteinuria (> 300 mg/24 h or Albustix +2) during the first, second and third term and 2-3 months after the delivery, as well as measurement of birth weight for determination of small for gestational age (SGA) babies, 37 women developed PIH, 10 without other complication, 16 with proteinuria (5 of which with SGA babies) and 11 with SGA babies. Furthermore 2 patients presented abruptio placentae without PIH. 395 women did not develop PIH including 389 normotensive women and 6 chronic hypertensive patients without superimposed toxemia. Only 1 was diabetic. None had chronic renal disease. Mean (+/- SD) levels of beta HCG were higher in PIH than in controls: 46,805 +/- 19,068 versus 23,479 +/- 13,463 IU. A pathologic threshold was chosen as the mean for the whole population + 1 SD: 25,613 + 15,479 = 41,082 IU. Elevated levels (above this value) were significantly associated with isolated PIH or PIH complicated with proteinuria and/or with SGA babies. The positive predictive value of this criterion was respectively 11, 15 and 12% for each of these complications. The relative risk (and 95% confidence limit) of women with elevated beta HCG for each of these complications was 20 (6-79), 11 (4-43) and 22 (7-93). Elevated plasma beta HCG found around 17 weeks of amenorrhea predicts PIH complicated with either proteinuria or SGA babies with a positive predictive value comparable to that of the best and earliest test proposed up to now to select nulliparas at high risk of preeclampsia, namely the abnormalities of the Doppler waveforms of the uterine arteries. Since this test is simpler to perform, it represents the most convenient method to screen a population of nulliparas for evaluation of the benefits of low-dose aspirin. PMID- 8730422 TI - Genetic predisposition to Balkan endemic nephropathy. AB - This study is the first cytogenetic investigation of healthy relatives of patients with Balkan endemic nephropathy (BEN) who were born in nonendemic areas. Characteristics of BEN No. 3 chromosomal anomalies (extremely high frequencies of 3q25 homologue discordance - 68.5 +/- (SD) 5.03% vs. 6.65 +/- 0.95% in controls, p < 0.001; chromosome breaks at 3q25 band - 0.79 +/- 0.25% vs. 0.01% in controls, p < 0.001; structural aberrations affecting 3q25 band), very high frequency of acquired chromosomal aberrations (5.74 +/- 0.64% vs. 1.72 +/- 0.3% in controls) and a family history with 1 or 2 BEN parents were identified in 5 relatives. It is proposed that they are at high risk for developing the disease and that a genetic mechanism might be involved in the etiology of BEN. PMID- 8730423 TI - Intracellular calcium signalling and vascular reactivity in Bartter's syndrome. AB - We investigated patients affected by Bartter's syndrome in the attempt to localize the intracellular defect mediating the reduced intracellular Ca2+ mobilization that may be responsible for the decreased vascular reactivity characteristic of Bartter's syndrome. Using the formylmethionyl-leucyl phenylalanine (fMLP) receptor system, which causes, intracellular calcium release, we investigated fMLP-stimulated intracellular inositol 1,4,5 trisphosphate (IP3) production as well as the number and affinity of fMLP receptors in neutrophils from Bartter's syndrome patients and healthy controls. Scatchard plot analysis of radioactive fMLP binding to neutrophils indicated that there were no differences in either cell receptor number and affinity for ligand between healthy controls (n = 5) and patients with Bartter's syndrome (n = 5): 6,151 +/- 1,431 vs. 7,112 +/- 2,566 receptors/cell; K(D): 0.446 +/- 0.14 vs. 0.454 +/- 0.09 pM of fMLP. 5- and 10-second fMLP-stimulated intracellular IP3 production was instead reduced in patients affected by Bartter's syndrome: 2.479 +/- 1.07 vs. 4.073 +/- 1.04 nmol/10(7) cells at 5 s (n = 8; p < 0.01); 1.673 +/- 0.741 vs. 3.766 +/- 1.348 nmol/10(7) cells at 10 s (n = 8; p < 0.005). The results of this study indicate that the anomaly of intracellular calcium mobilization in patients with Bartter's syndrome arises from a defect at the postreceptor level. The anomalous calcium signalling that takes place in Bartter's syndrome may provide a mechanism for the hyporesponsiveness to pressor stimuli characteristic of these patients. PMID- 8730424 TI - Correlation between serum carnitine levels and erythrocyte osmotic fragility in hemodialysis patients. AB - The relationship between serum carnitine levels and erythrocyte osmotic fragility was investigated in 26 chronic hemodialysis patients (10 males and 16 females, mean age: 57.3 +/- 13.5 years). Serum total-carnitine (TC), free-carnitine (FC) and acyl-carnitine (AC) levels were determined by a spectrophotometric method. Erythrocyte osmotic fragility was measured with a coil planet centrifuge. Serum TC levels were 39.9 +/- 13.4 mumol/l (mean +/- SD), FC levels were 21.8 +/- 7.8 mumol/l and AC levels were 18.0 +/- 9.6 mumol/l. The mean hemolysis end point (HEP) was 67.4 +/- 5.4 mOsM, the hemolysis maximum point (HMP) was 86.3 +/- 5.4 mOsM and the hemolysis start point (HSP) was 101.2 +/- 4.4 mOsM. Each hemolysis point in hemodialysis patients was elevated in comparison with the normal range. There were no significant differences in hemolysis points between a recombinant human erythropoietin (rhEPO)-treated group and nontreated group. HEP correlated with serum TC (r = -0.56, p < 0.01) and AC levels (r = -0.58, p < 0.01). HMP correlated with serum TC (r = -0.42, p < 0.05) and FC levels (r = -0.41, p < 0.05). Dose requirement of rhEPO maintaining target hematocrit correlated with serum TC (r = 0.54, p < 0.05) and FC levels (r = 0.50, p < 0.05). These data support that low serum carnitine levels accelerate erythrocyte osmotic fragility. Carnitine may contribute to the metabolism of erythrocyte membrane and have an impact on the efficacy of rhEPO in correcting renal anemia. PMID- 8730425 TI - Tenascin is an ubiquitous extracellular matrix protein of human renal interstitium in normal and pathologic conditions. AB - Tenascin, a large oligomeric glycoprotein, is a recent addition to a list of increasing extracellular matrix proteins. Previous studies have documented the strong expression of tenascin in embryonic kidney and in both normal and abnormal mature glomeruli implicating an important role of this extracellular matrix protein in nephrogenesis and glomerular scarring. Whether tenascin plays any role in interstitial fibrosis, a common final pathway of tubulointerstitial nephritis, is no known; on the other hand, a detailed knowledge of the structural components of interstitial fibrosis is essential for further studies on other fundamental aspects of this biologically and clinically important process. In this study, the expression of tenascin in the renal interstitium was immunohistochemically evaluated in 208 renal specimens during normal kidney (23 cases), acute tubular necrosis (8), acute tubulointerstitial nephritis (8), chronic primary tubulointerstitial nephritis (30), tubulointerstitial nephritis secondary to glomerular diseases of mild (46) and severe (55) degree, ischemic damage (24), and rejection (14). It was found that in normal kidney tenascin expression was limited to the medullary interstitium. In kidney with tubulointerstitial nephritis, tenascin was ubiquitously and constantly expressed in any areas with tubulointerstitial damage regardless of diagnosis, etiology, the cortical vs. medullary location of the lesions, stage of the fibrogenetic process, density of fibroblasts, or severity of interstitial inflammation in the affected areas. Indeed, strong tenascin expression was seen in areas where there was only interstitial edema or inflammation as judged by routine light microscopic preparations. In summary, this study systematically documents tenascin as a novel extracellular matrix protein selectively expressed in the medullary interstitium in normal kidney, and ubiquitously present in areas with interstitial fibrosis. PMID- 8730426 TI - Patch-clamp study on T-lymphocyte potassium conductance in patients with chronic renal failure. AB - Using the patch-clamp technique, we studied the differences in whole-cell potassium conductance (g(K)+) in T lymphocytes (TL) from three groups of patients suffering from renal failure: not dialyzed patients, dialyzed patients, and dialyzed patients treated with human recombinant erythropoietin (rHuEPO). The differences in g(K+) values in the group of not dialyzed patients in comparison with controls was not significant (p > 0.05). In the group of dialyzed patients, after roughly 6 years of the hemodialysis therapy, the g(K+) value was significantly higher than in controls. In dialyzed patients treated with rHuEPO, g(K+) value was significantly lower in comparison with control. Moreover, in dialyzed patients treated with rHuEPO, the time duration of dialysis therapy did not significantly affect the TL whole-cell conductance. We conclude that the g(K+) is changed in TL in renal failure patients and that the time duration of hemodialysis therapy as well as the use of rHuEPO affect the g(K+) value. Possible mechanisms underlying the observed changes in g(K+) values, as well as medical implications of obtained results are discussed. PMID- 8730427 TI - Genetic studies in a black family with autosomal dominant polycystic kidney disease and sickle-cell trait. AB - Autosomal dominant polycystic kidney disease (ADPKD) is caused by at least two different genes. The ADPKD1 gene is located on chromosome 16p and a second locus is at 4q. Although the ADPKD1 gene is responsible for the majority of the disease in whites, there was no information regarding the gene type in blacks. We studied a black family which presented with both ADPKD and sickle-cell trait (SA) to determine which ADPKD gene was present in this family, and to examine linkage between the ADPKD in this family and markers for the beta-hemoglobin gene on chromosome 11. The ADPKD in this family was linked to markers on chromosome 16, and no linkage was found with the beta-hemoglobin gene. Family members with SA and ADPKD had an early onset of end-stage renal disease. The hemoglobin haplotype was identified as the Central African Republic-type, which has been reported to be associated with a higher incidence of renal failure in sickle-cell anemia. PMID- 8730428 TI - Failure of human immunoresponse to N-glycolylneuraminic acid epitope contained in recombinant human erythropoietin. AB - Recombinant human erythropoietin (rHUEPO) was produced by Chinese hamster ovary cells and commercially distributed to hospitals by two pharmaceutical companies in Japan ('ESPO' by Kirin Brewery Co. Ltd., and Sankyo Co. Ltd., and 'EPOGIN' by Chugai Pharmaceutical Co. Ltd.) These products contained about 1% N glycolylneuraminic acid (Neu5Gc) in total sialic acid content. Since humans do not synthesize Neu5Gc, successive injection of Neu5Gc-containing products was feared to lead to allergic-like symptoms. Therefore, serum levels of antibodies of Neu5Gc epitope in 90 patients who received repeated i.v. injections of ESPO or EPOGIN were determined by an enzyme immunoassay using Neu5Gc alpha 2-3Gal beta 1 4Glc-Cer, GM3(Neu5Gc), as an antigen and compared with those in 100 healthy persons. Either no or low antibody levels were detected in both groups with no significant difference. In 40 patients who received s.c. injections of ESPO or EPOGIN, serum HD antibody levels were determined before and after weekly therapeutic injections carried out for one to several months, but no significant elevations were detected in all patients. The above results indicated that therapeutic administration of rHuEPO to patients to patients with chronic renal failure is safe from allergic-like side effects associated with the production of Neu5Gc-specific antibodies, and it was concluded that Neu5Gc epitope of rHuEPO is minimally antigenic in humans. PMID- 8730429 TI - Prevalence of endemic distal renal tubular acidosis and renal stone in the northeast of Thailand. AB - We have previously reported a large group of patients with endemic distal renal tubular acidosis (EdRTA) admitted to the hospitals in the northeast of Thailand. Since large number of patients were identified in a relatively short period of time, and in an area whose population is homogeneous, we were led to investigate the prevalence of the condition in the area. A survey was conducted in five villages (total population of 3,606) within the northeast of Thailand. 3,013 villagers were examined for urinary citrate concentration and short acid loading test was performed in those with low urinary citrate. 2.8% of the population (2.2 3.4%, 95% confidence interval) failed to lower their urine pH after acid loading; within this group, 0.8% of the population had serum potassium less than or equal to 3.5 mEq/l. In addition a large number of villagers were found to have low urinary citrate concentration and there was concurrent high prevalence of renal stone. The prevalence of EdRTA and renal stone was higher in villagers with poorer socioeconomic status, suggesting that environmental factors play a major role in their pathogenesis. Villagers with acidification defect have 2.4 times the chance of having renal stone and/or nephrocalcinosis. EdRTA is therefore one of the important factors responsible for the high prevalence of renal stone in the area. In conclusion we have confirmed the high prevalence of EdRTA in the northeast of Thailand and provided data showing high prevalence of renal stone and hypocitraturia in the same population. PMID- 8730430 TI - Platelet-activating factor levels and PAF acetylhydrolase activities in patients with primary glomerulonephritis. AB - Platelet-activating factor (PAF) belongs to a new class of lipid chemical inflammatory mediators described as acetyl glyceryl ether phosphorylcholine. Although PAF has been implicated in kidney injury, its role in renal immune injury has not been clearly defined yet. We studied the levels of PAF in the plasma and urine and acetylhydrolase (AH) activities in serum and in renal tissue (cortex, C and medulla, M) in patients with primary glomerulonephritis (PGN). PAF levels in the plasma and urine and AH activity in serum of normal volunteers as well as AH activities in normal renal parenchyma (C and M) from nephrectomized patients (served as control) were also measured. Our results demonstrate increased PAF levels in the plasma and urine as well as increased AH activity in serum in patients with PGN in comparison to normal volunteers. AH activity in cortex of those patients was diminished compared to normal kidney tissue. We propose that the enhanced AH activity in serum in patients with PGN could be due to hyperproduction and low degradation of PAF in nephritic tissue which on one hand results in enhanced PAF levels in the plasma and in urine and on the other hand results in enhanced serum AH activity by virtue of substrate (PAF) availability. These data provide new knowledge in the homeostasis of PAF and its degrative enzyme in the setting of PGN. PMID- 8730431 TI - Hemodialysis-related pruritus: a double-blind, placebo-controlled, crossover study of capsaicin 0.025% cream. AB - Pruritus is a significant symptom among patients receiving hemodialysis. However, its underlying mechanisms remain obscure. Substance P, a neuropeptide, has been implicated in the mediation of pain and some itch sensations. Local application of capsaicin depletes the peripheral neurons of substance P and may block the conduction of pain or pruritus. This study aims to assess the efficacy and safety of capsaicin 0.025% cream in the treatment of hemodialysis-related pruritus and to further explore the underlying pathomechanism. Nineteen hemodialysis patients with idiopathic, moderate (n = 5) to severe (n = 14) pruritus were examined in a double-blind, placebo-controlled, crossover study and 17 of them completed the study. Topical agent of capsaicin or placebo base cream was applied to localized areas of pruritus 4 times a day. The severity of pruritus and treatment-related side effects (cutaneous burning/stinging sensations, dryness, or erythema) were evaluated weekly. The results showed (1) that 14 of 17 patients reported marked relief and 5 of these 14 patients had complete remission of pruritus during capsaicin treatment (Wilcoxon signed-ranks test, 2p < 0.001); (2) capsaicin was significantly more effective than placebo (Mann-Whitney rank sum test, 2p < 0.001) and a prolonged antipruritic effect was observed 8 weeks posttreatment; (3) no serious side effects were noted during the study and (4) there were no significant changes in serum concentrations of albumin, calcium, phosphorus, alkaline phosphatase, or intact parathyroid hormone during the treatment with either capsaicin or placebo. In summary, the present study indicates indirectly that idiopathic pruritus in some patients on maintenance hemodialysis may be transmitted by substance P from the peripheral sensory neurons to the central nervous system. Topical capsaicin with the unique pharmacological effect is demonstrated to markedly improve the pruritus of these patients. PMID- 8730432 TI - Regulation of the C3 production by gamma interferon from peripheral blood T cells in patients with membranoproliferative glomerulonephritis and poststreptococcal acute glomerulonephritis. AB - This study assessed the regulatory role of peripheral blood T cells in the C3 production in patients with poststreptococcal glomerulonephritis and branoproliferative glomerulonephritis. Peripheral blood T cells from patients at various stages of disease were cultured and the supernatants tested for gamma interferon (IFN-gamma) content and the capacity to stimulate C3 production by HuH 7 cells. Supernatants from patients with membranoproliferative glomerulonephritis and from convalescent patients with poststreptococcal glomerulonephritis significantly stimulated the C3 production; the degree of stimulation correlated with the IFN-gamma content of the supernatants. Similar results were obtained using recombinant IFN-gamma. In both cases, the effect was blocked by the addition of anti-IFN-gamma monoclonal antibody to the cultures. Interleukin 2 and interleukin 6 levels in supernatants from T cell cultures of patients and controls were essentially the same. In summary, IFN-gamma plays a regulatory role in C3 production by human hepatoma cell lines. PMID- 8730433 TI - Blockade of radiocontrast-induced nephrotoxicity by the endothelin receptor antagonist, SB 209670. AB - The effect of the novel nonpeptide endothelin (ET) receptor antagonist, (+/-)-SB 209670, on the renal effects of the high osmolar tri-iodinated ionic contrast media, Hypaque, was evaluated in anesthetized dogs in the presence or absence of the cyclooxygenase inhibitor, indomethacin (10 mg/kg, i.v.). Hypaque alone (5 ml/kg, i.v.) resulted in a marked diuresis and natriuresis but little change in either renal blood flow or renal vascular resistance. When the ET receptor antagonist, (+/-)-SB 209670, was infused into the renal artery at a dose that inhibited ET-induced renal vasoconstriction, Hypaque resulted in a significant increase in renal blood flow and decrease in renal vascular resistance. In the presence of indomethacin, Hypaque caused a significant increase in renal vascular resistance which was abolished by (+/-)-SB 209670. The data indicate that the radiocontrast media, Hypaque, can cause renal vasoconstriction which may be mediated by ET. PMID- 8730434 TI - The influence of oxygen free radical scavengers on the reduction of membrane bound Na(+)-K(+)-ATPase activity induced by ischemia/reperfusion injury in the canine kidney. AB - The present study was designed to determine whether the administration of free radical scavengers, superoxide dismutase (SOD), catalase or dimethylsulfoxide (DMSO) is able to ameliorate ischemia/reperfusion injury in the canine kidney and also ascertain whether or not a relationship exists between oxygen free radicals and membrane-bound Na(+)-K(+)-ATPase activity. In 23 dogs, the vascular pedicle of the left kidney was clamped for 75 min at room temperature. The experimental animals received free radical scavengers for 30 min starting at 2 min prior to reperfusion. Renal tissue specimens were enzyme-histochemically examined regarding the activity of membrane-bound Na(+)-K(+)-ATPase, and a marked reduction just before reperfusion was revealed. The SOD- and the DMSO-treated groups showed a marked recovery of the membrane-bound Na(+)-K(+)-ATPase activity; however, the untreated and the catalase-treated groups still demonstrated a marked reduction 1 day after reperfusion. At the same time, widespread acute tubular necrosis in the cortex was observed in the untreated and catalase groups in comparison with the SOD and the DMSO groups. In addition, the SOD and the DMSO groups significantly preserved better renal function. Based on these findings, it was thus concluded that free radical scavengers ameliorate the recovery of depressed membrane-bound Na(+)-K(+)-ATPase activity and ischemia/reperfusion injury in the canine kidney. PMID- 8730435 TI - Renin immunochemistry, sodium excretion and relative heart weight in cyclosporine or alimentary-induced magnesium deficiency in rats. AB - Rats were given a magnesium-(Mg) depleted (Mgd), or a Mg-standard (Mgst) or a Mg enriched (Mge) diet, with 20 mg/kg/day cyclosporine (Cy) or olive oil per os for 90 days (6 groups). Anti-renin antibody was applied and the percent of renin positive glomeruli (RI) was taken. Sodium excretion (NaU), relative heart weight (HW), as a measure of hypertension, and total femur Mg were measured. Compared to dietary controls, femur Mg was reduced under Cy and Mgd or Mgst indicating Mg deficiency. RI was higher in all Cy groups (p < 0.01), and Nau was lower in Mgd + Cy and in Mgst + Cy (p < 0.01). Correspondingly, HW was found to be significantly higher in Mgd + Cy and Mgst + Cy. In animals under Mge + Cy, there were no differences in NaU and HW compared to controls. The results indicate a relation between Cy-related hypertension and Mg status: Mg deficiency seems to enhance the hypertensive effect of Cy via sodium retention. PMID- 8730436 TI - Increased gene expression of insulin-like growth factor-i receptor in experimental diabetic rat glomeruli. AB - Renal hypertrophy is a characteristic and early manifestation of diabetes in humans and experimental animals. We examined the precise distribution of insulin like growth factor-I (IGF-I) mRNA and IGF-I receptor mRNA in the experimental diabetic rat kidney using a nonradioactive in situ hybridization technique. No significant difference in the distribution of IGF-I mRNA was found between the diabetic and control rats. IGF-I mRNA-positive cells were found in the collecting ducts and in scattered single cells in the distal tubules. The number of IGF-I mRNA-positive cells was very low in the glomeruli. Expression of IGF-I receptor mRNA was rarely seen in the glomeruli of control rats. IGF-I receptor mRNA was detected after induction of diabetes in glomerular mesangial, visceral epithelial, and parietal epithelial cells. The number of IGF-I receptor mRNA positive cells in a glomerulus increased significantly, peaking at 4 weeks as compared with the control rats. Overexpression of IGF-I receptor in glomerular cells, especially mesangial and visceral epithelial cells, may contribute to glomerular hypertrophy in diabetic nephropathy. PMID- 8730437 TI - Anemia of chronic renal failure: characterization in the mouse and correction with human recombinant erythropoietin. AB - Anemia is a cardinal feature of chronic renal failure (CRF) which contributes significantly to the clinical syndrome of chronic uremia. We have conducted a detailed examination of the hematological changes in CRF in the inbred mouse strain C57BL/6J. As in the human situation, CRF mice presented major hematological changes affecting primarily the erythroid cell series. Despite the presence of abundant iron stores in the bone marrow, the CRF mice developed a hypoproliferative anemia of a severity commensurate with the degree of renal impairment. The levels of circulating erythropoietin (EPO) in CRF mice were not significantly different from those in normal control littermates and were therefore inappropriately low for the degree of anemia. In contrast acutely bled control mice with normal renal function showed a significant inverse correlation between the serum EPO level and hemoglobin concentration, indicating an appropriate response to anemia. The chronic administration of recombinant human EPO raised the hemoglobin concentration of CRF mice, a therapeutic effect which was independent of the initial degree of anemia. These observations suggest that this animal model has wide applicability for the study of anemia secondary to CRF. PMID- 8730438 TI - Localization of glutamic acid decarboxylase in the kidneys of nonobese diabetic mice. AB - Antibodies to glutamic acid decarboxylase (GAD), a pancreatic islet beta-cell antigen, are present in > 80% of newly diagnosed insulin-dependent diabetes mellitus (IDDM), and are found in nonobese diabetic (NOD) mice, a murine model of spontaneous IDDM. To determine whether GAD is a target antigen in the kidney damage of NOD mice, we studied GAD mRNAs (GAD65 and GAD67) by RT-PCR in mesangial cells, isolated glomeruli, and kidney cortex and medulla in NOD and SJL/C57BL mice. GAD mRNAs were detected in the cortex of both diabetic and nondiabetic NOD and SJL/C57BL mice and GAD antigen was present in proximal and distal tubules by immunofluorescence microscopy. Neither GAD antigen nor mRNA were present in mesangial cells or glomeruli of diabetic or nondiabetic mice. Thus, the expression of GAD in renal tubules raises the possibility that GAD antigens may play a role in diabetic tubulointerstitial disease, whereas the absence of these antigens in glomeruli suggests that GAD-triggered autoimmunity is not directly involved in the glomerular lesions. PMID- 8730439 TI - Evidence for the involvement of vasopressin in the pathophysiology of adriamycin induced nephropathy in rats. AB - The effect of orally available, nonpeptide vasopressin V1 and V2 receptor antagonists on chronic progressive glomerular disease was investigated in Wistar rats with Adriamycin-induced nephropathy. At weeks 0 and 3, Adriamycin was injected twice, and at week 3 drugs started to be given as follows: groups 2 and 3 were treated with V1 and V2 antagonists, respectively, while the untreated group 1 served as control. To block the effects of vasopressin totally, both V1 and V2 antagonists were simultaneously administered (group 4). At weeks 8 and 10, V1 and V2 antagonists given either alone or combined significantly reduced the urinary protein excretion to the same levels. Urinary volume increased in groups 3 and 4 from week 4. Systolic blood pressure did not significantly increase in all groups during the study. Histological alterations in the kidney of groups 2, 3 and 4 were significantly attenuated compared to the control. These results suggest that both vasopressin V1 and V2 agonism plays a role in the pathophysiology of Adriamycin-induced nephropathy despite plasma levels of vasopressin within the normal range. These findings also lead to the notion that in some types of nephrotic patients these orally available V1 and/or V2 receptor antagonists may be effective for reduction of proteinuria and for retardation of progression of renal failure. PMID- 8730440 TI - Radial artery pseudoaneurysm complicating a Brescia-Cimino dialysis fistula. AB - The authors describe an unusual case of radial artery pseudoaneurysm in a patient with a Brescia-Cimino fistula. Evaluation of a pulsatile mass on the patient's forearm which occurred after hemodialysis led to a nondiagnostic fistulogram. Color flow ultrasound was helpful in identifying the nature of the abnormality, which was subsequently confirmed arteriographically. This case illustrates that fistulography is not 100% sensitive and should be supplemented with other imaging modalities when the clinical and fistulographic findings are discordant. PMID- 8730441 TI - Rhabdomyolysis and acute renal failure following blowpipe dart poisoning. AB - The latex of pokok ipoh (Antiaris toxocaria) and the root bark of akar ipoh (Strychnos species) have been the main sources of the poisonous principles in dart and arrow poisons prepared throughout south-east Asia. We report a fatal case of rhabdomyolysis and acute oliguric renal failure following oral ingestion of blowpipe dart poison. To our knowledge this is the first such report. PMID- 8730442 TI - Acne: a potential side effect of cyclosporine A therapy. AB - A renal transplant recipient who developed severe acne 6 months after transplantation is described. Maintenance immunosuppression consisted of cyclosporine A (CsA), azathioprine and prednisone. Tapering the prednisone dose to as low as 5 mg/day, in addition to topical tetracycline, Retin-A cream, and systemic antimicrobial therapy failed to control the progression of the skin lesions. Despite therapy with isotretinoin (Accutane), the lesions continued to progress with nodulocystic transformation (acne conglobata) and isotretinoin was discontinued after 4 months. However, the condition continued to worsen with the development of a systemic illness with daily fever, diaphoresis, and depression. High fever (103 degrees F) with shaking chills prompted hospitalization. Withdrawal of CsA resulted in rapid and continuous improvement of the skin lesions. After 12 months of follow-up, the lesions significantly resolved except for residual areas of scarring. No episodes of acute allograft rejection occurred. In conclusion, we suggest that CsA therapy may be associated with the development of acne. Nodulocystic transformation (acne conglobata) may occur despite the use of isotretinoin. Finally, withdrawal of CsA may lead to resolution of the skin disease and should, therefore, be considered as a therapeutic option for severe and treatment-resistant cases. PMID- 8730443 TI - Myelofibrosis secondary to renal osteodystrophy. AB - A hemodialyzed women with secondary hyperparathyroidism who recovered well from myelofibrosis after a total parathyroidectomy with autotransplantation of parathyroid tissue to the forearm (PTx) is described. Before the operation, she had received regular transfusions to maintain an adequate hematocrit even under recombinant human erythropoietin (rhEpo) therapy. She showed splenomegaly and leukoerythroblastosis was present in her peripheral blood. A bone marrow biopsy and bone marrow scintigraphy confirmed the diagnosis of myelofibrosis. After PTx, her hematocrit gradually increased without any transfusion. It has been maintained around 35% now 16 months since the operation. Her spleen has also gradually decreased in size. In addition, no leukoerythroblastosis has been found in the peripheral blood. Serial follow-up scintigraphy of bone marrow revealed a decline in extramedullary hematopoiesis. These findings indicated that her myelofibrosis was the result of secondary hyperparathyroidism, and that this complication is potentially reversible if accurate treatment is given. Physicians dealing with the end-stage renal disease should be aware of this complication to avoid additional transfusions. PMID- 8730444 TI - Acute brucellosis associated with massive proteinuria. AB - Mild proteinuria during these renal complications in brucellosis is a common feature, however, heavy proteinuria in association with renal brucellosis has rarely been reported and it always was associated with a concomitant development of renal failure. This is the first description of heavy proteinuria as the sole renal derangement during acute infection with brucellosis. PMID- 8730445 TI - Tuberculosis-associated hemophagocytic syndrome in a hemodialysis patient: case report and review of the literature. AB - We present an unusual hematologic manifestation of tuberculosis in a hemodialysis patient. A 37-year-old man was admitted because of fever, anorexia, and weight loss which developed 3 months earlier. He previously received a renal transplant and commenced hemodialysis 1 year ago. Physical examination revealed hepatomegaly, and peripheral blood counts revealed pancytopenia. The delayed bone marrow biopsy demonstrated marked hemophagocytosis, granuloma and tubercle bacilli by the acid-fast stain. The clinical course was fatal and he died without definite treatment. Through this case, we would like to recommend early bone marrow biopsy and antituberculosis therapy if fever of unknown origin, anorexia, weight loss, hepatomegaly and pancytopenia develop in patients undergoing dialysis. PMID- 8730446 TI - Hyperammonemia in a patient with short bowel syndrome and chronic renal failure. AB - A patient with short bowel syndrome (SBS) and renal failure developed a disturbance of consciousness with hyperammonemia. Abnormally low concentrations of ornithine, citrulline, and arginine were observed on the plasma aminogram. These results suggested that the activities of amino acid synthetase localized in the small intestinal flora were lost. The small intestine is required for arginine synthesis; thus, infusion limited to the essential amino acids to SBS patients will cause a deficiency of the urea cycle intermediates, ornithine, citrulline, and arginine and may lead to hyperammonemia. In addition, the renal insufficiency may have caused decreased excretion of ammonia. In this patient, supplemental arginine improved the symptoms. PMID- 8730447 TI - Use of a phosphorus-enriched dialysate in a hypercatabolic renal failure patient receiving intensive hemodialysis therapy. AB - An elderly man with end-stage renal disease required intensive hemodialysis therapy because of sepsis-induced hypercatabolism. We were able to prevent the occurrence of hypophosphatemia by using a phosphorus-enriched dialysate during hemodialysis treatments. PMID- 8730448 TI - A case of renovascular hypertension associated with neurofibromatosis. AB - We report a case of renovascular hypertension associated with neurofibromatosis complicated by moderate proteinuria. A 16-year-old female was admitted to Kensei General Hospital with a complaint of headache and a blood pressure of 230/120 mm Hg. She was referred to us for further evaluation of the hypertension. On examination, cafe-au-lait spots were seen over her extremities and flank, and a bruit was heard in the right upper abdomen. The urinary protein excretion was 2.1 g/day. The plasma renin activity (PRA) and plasma aldosterone concentration were high, but the levels of catecholamines were normal. The renogram was asymmetric and on venous sampling, the PRA in the right renal vein was 58.3 ng/ml/h and that in the left was 22.1 ng/ml/h. CT scan detected an approximately 10-mm mass in the proximal right renal artery. Arteriography disclosed severe stenosis in the right renal artery and the superior mesenteric artery. Therefore, we concluded that her hypertension resulted from stenosis of the right renal artery due to neurofibromatosis. Accordingly, she underwent an operation to reconstruct that artery. After the operation, her blood pressure and PRA normalized without administration of any anti-hypertensive drug and urinary protein disappeared. PMID- 8730449 TI - Posttraumatic chyluria due to lymphorenal fistula regressed after somatostatin therapy. AB - A sudden-onset chyluria after trauma was evaluated giving evidence of a lymphatic urinary fistula in the right kidney. Treatment with somatostatin normalized the urinary pattern and the result was maintained even after the discontinuation of the therapy. PMID- 8730450 TI - Fibroadhesive form of tuberculous peritonitis: chyloperitoneum in a patient undergoing automated peritoneal dialysis. AB - Chyloperitoneum is a rare condition in patients undergoing peritoneal dialysis. We report here a patient who developed chylous ascites during the course of tuberculous peritonitis. The diagnosis was confirmed by cultures of dialysate and peritoneal biopsy, and laparoscopy revealed severe hyperemia and intestine adhesion. Intrinsic lymphatic obstruction and superimposed peritoneal fibrosis together might be responsible for the pathogenesis of this special presentation. Although in most cases of continuous ambulatory peritoneal dialysis, the causes of chyloperitoneum remained unknown, we suggest, from the experience of this case, that tuberculous peritonitis, especially the fibroadhesive form, should be highly suspected in any dialysis cases with chyloperitoneum. Laparoscopy should be initiated early, particularly when the culture is negative for common pathogens or when the patient responds poorly to the usual antimicrobial agents. PMID- 8730451 TI - Serum lipoprotein (a) concentration is increased in moderate renal failure. PMID- 8730452 TI - Effect of erythropoietin administration on thyroid functions of the patients undergoing regular hemodialysis. PMID- 8730453 TI - Platelet glycoproteins GPIb and GPIIb/IIIa abnormalities in uremia. PMID- 8730454 TI - Significance of serum fibrin/fibrinogen degradation products measured by a highly sensitive ELISA method in renal diseases. PMID- 8730455 TI - Parvovirus B19 and hemodialysis. PMID- 8730456 TI - Hepatitis C virus infection in patients on peritoneal dialysis, hemodialysis and in dialysis staff members in south Serbia. PMID- 8730457 TI - Contamination by hepatitis C in a haemodialysis center: preventive measures. PMID- 8730458 TI - Efficiency of once-weekly subcutaneous administration of recombinant human erythropoietin versus three times a week administration in hemodialysis patients. PMID- 8730459 TI - Helicobacter pylori antibodies in patients on chronic hemodialysis. PMID- 8730460 TI - Human immunodeficiency virus infection in patients on maintenance dialysis. PMID- 8730461 TI - Acute effect of erythropoietin on endothelin release in uremia. PMID- 8730462 TI - Localization of beta-2-microglobulin in prostatic corpora amylacea of prostatic hypertrophy patients. PMID- 8730463 TI - Convulsions, hypertension crisis and acute renal failure in postpartum: role of bromocriptine? PMID- 8730464 TI - Rapidly progressive glomerulonephritis following hemolytic crisis. PMID- 8730465 TI - Soluble ELAM-1 is elevated with the progression of IgA nephropathy but not with that of polycystic kidney disease. PMID- 8730466 TI - Ambulatory blood pressure monitoring in chronic haemodialysis children with end stage renal failure. PMID- 8730467 TI - Glomerular deposition of hepatitis C virus in membranoproliferative glomerulonephritis. PMID- 8730468 TI - Biology and genetic heterogeneity of hepatitis C virus. AB - Hepatitis C virus (HCV) has a significant, albeit varied, presence around the world. This virus is primarily transmitted parenterally, although sexual and perinatal transmission does appear to occur. However, no risk factor for transmission could be identified in a significant proportion of infected individuals. It was found that individuals became viremic early after the primary HCV infection, whereas seroconversion and hepatitis occurred several weeks later. It was demonstrated that less than 20% of patients cleared their viremia, with the majority of patients becoming chronically infected. A significant proportion of chronically infected individuals developed chronic hepatitis and liver cirrhosis, and a strong association has been found with the development of hepatocellular carcinoma. Finally, HCV seems to be associated with autoimmune diseases and type II cryoglobulinemia. Thus, significant morbidity and mortality is caused by HCV infection worldwide. In a single infected individual the genome population of HCV has been found to comprise a quasispecies that consists of a number of identical sequences (i.e., the master sequence) and other closely related sequences. The master sequence of this quasispecies population changes during infection. In particular, it has been found that the sequence of the hypervariable region I changes rapidly in infected individuals. It is possible that the quasispecies nature of HCV constitutes a mechanism by which HCV escapes immune surveillance and establishes a persistent infection in the host. It is now well established that HCV exists as distinct genotypes among different HCV isolates. According to the currently used classification these can be divided into a number of major genetic groups (or types) and subgroups (or subtypes). The extensive genetic heterogeneity of HCV has important implications for diagnosis, pathogenesis, treatment and vaccine development. PMID- 8730469 TI - Immunity elicited by hepatitis C virus. AB - Hepatitis C virus (HCV), the major causative agent of post-transfusion and community-acquired non-A, non-B (NANB), is a single-stranded RNA virus characterized by a high degree of genetic heterogeneity. HCV is endemic worldwide and is a major cause of chronic liver disease and hepatocellular carcinoma. The development of a broadly reactive vaccine is a high priority for the control of HCV infection. In recent years, however, serious concerns have been raised regarding the degree of protective immunity elicited by HCV in the host. Several observations, both in patients and in the chimpanzee model, have suggested a lack of protective immunity against HCV. Chronic HCV infection develops in more than 80% of patients, suggesting that in most cases the immune response of the host fails to mediate resolution of the infection. Cross-challenge studies demonstrated that convalescent chimpanzees are not protected against re-infection with homologous or heterologous HCV strains. Similar evidence has been obtained in polytransfused beta-thalassemic children, in whom re-infection with HCV was associated with multiple episodes of acute hepatitis. Although most of the evidence thus far accumulated suggests that HCV does not elicit a protective immune response, recent studies have provided experimental evidence, both in vitro and in vivo, that HCV infection induces a neutralizing antibody response in humans. However, such antibodies are isolate-restricted and ineffective against variant HCV strains emerging in vivo. Recently, using recombinant envelope proteins of HCV, a successful vaccination of chimpanzees against challenge with a homologous viral strain was reported. Whether this vaccine can provide protection against challenge with a higher infectious dose of the homologous virus or against challenge with heterologous strains of HCV remains to be established. Overall, the data hitherto accumulated indicate that the genetic heterogeneity of HCV will be a major impediment for the development of a broadly reactive vaccine for the control of HCV infection. PMID- 8730470 TI - Hepatitis type C: modes of transmission and preventive measures. AB - There is general agreement that the hepatitis C virus is efficiently transmitted parenterally, while data on viral transmission from mothers to babies or by sexual or non-sexual household contact are conflicting. In Italy, hepatitis C accounts for approximately 20% of all cases of acute viral hepatitis. It is more frequent among young persons (15-24 years old), among those living in the southern part of the country or on the islands, and in males than in females (3:1). In recent years the incidence rate has dropped and significant changes in the HCV transmission patterns have been observed. As a consequence of the implementation of anti-HCV screening and the introduction of increasing restrictions on donor eligibility, the proportion of cases arising from blood and blood component transfusions has significantly declined. A candidate vaccine to protect against hepatitis C virus is now under development. PMID- 8730471 TI - Clinical evolution of HCV-related chronic hepatitis. AB - Evolution of HCV positive chronic hepatitis to cirrhosis has been reported to occur in about 30% of patients after five years of follow-up. In contrast, evolution of cirrhosis to decompensation and liver failure is slow, with a survival rate at 5 and 10 years of 91% and 79%, respectively. These findings support the hypothesis that histologic cirrhosis and clinical cirrhosis are different facets of the same disease, the latter developing after a long period of time and being related not only to liver disease per se, but also to other factors only partially identified. During this long-term evolution, extrahepatic manifestations may occur due to the formation of antigen-antibody immunocomplexes, which may eventually lead to death due to renal or cardiac complications. PMID- 8730472 TI - Hepatitis C virus and hepatocellular carcinoma. AB - Epidemiological evidence indicates that HCV plays an important role in the pathogenesis of many cases of hepatocellular carcinoma (HCC) in both hemispheres. In southern European countries the proportion of HCCs attributable to HCV is high, whilst in the USA this fraction is definitely smaller. These differences cannot be explained merely on the basis of the different prevalences of HCV in the corresponding general populations. Thought the mechanisms of HCV-related carcinogenesis are still obscure, it is clear that HCV alone or in conjunction with other environmental risk factors such as alcohol consumption or HBV infection can contribute to the epidemiological and clinical heterogeneity of HCC. Preliminary data indicate that HCV-related tumors arise in older patients, have a less aggressive course, and are more often associated with cirrhosis than HBV-related tumors. PMID- 8730473 TI - Immunohistochemical detection of hepatitis C virus-related proteins in liver tissue. AB - OBJECTIVE: Hepatitis C virus (HCV)-associated antigens (Ags) are hard to detect and poorly defined in liver tissue, and are of uncertain interpretation. The failure of immunohistochemistry in HCV infection may be due to the affinity of specific antisera, the levels of Ags in infected tissues, the labile and unstable expression of antigenic determinants, and the use of fixatives that may alter or destroy viral epitopes. Strategies to optimize all stages of tissue specimen processing have therefore been devised in the liver biopsies of patients with acute and chronic hepatitis, and those with hepatocellular carcinoma (HCC). METHODS: HCV-Ags were detected with a two-stage indirect immunostaining procedure on unfixed cryostat liver sections from 7 acute and 23 chronic HCV-infected patients, and from 4 patients with HCV-associated HCC. A mixture of monoclonal antibodies directed to structural and non-structural HCV-related proteins were used as the primary reagents. RESULTS: HCV-Ags in 50-70% of the hepatocytes were found in all seven acute hepatitis patients compared with < or = 20% hepatocytes (P < 0.05) in 10 out of 23 patients (43.5%) with chronic hepatitis. Immunoreactive signals appeared as diffuse or coarse granular deposits in the cytoplasm only. The nuclei were unstainable. No clear membranous pattern was found, although fine granular, submembranous accumulation in distinct areas of the cytoplasm was observed. In acute hepatitis, HCV-Ag positive hepatocytes were distributed in the lobules in direct relation to the areas of necrosis and inflammatory cell accumulation, whereas in chronic hepatitis the immunoreactive cells were not clearly related to the necrotic foci. HCV-Ag immunodeposits were demonstrated in all patients with HCC. The immunoreactive signal in neoplastic cells was primarily located in the cytoplasm and rarely in the nuclei. As compared with the non-neoplastic zones, neoplasia demonstrated a significantly higher specific signal. CONCLUSIONS: Immunohistology is a powerful tool for the identification of HCV-related proteins in liver tissue. Sensitivity was significantly enhanced by the use of fresh-frozen tissues, which presumably preserve their HCV antigen structure, and by a mixture of monoclonal antibodies directed against HCV-related proteins, possibly on account of the separate access of each probe to different target proteins. The demonstration of HCV infection in hepatocyte cytoplasm indicates that this is the primary site of HCV replication, while its presence in malignant cells suggests that the virus could be substantially involved in the pathogenesis of HCC. PMID- 8730474 TI - Hepatitis C virus as a lymphotropic agent: evidence and pathogenetic implications. AB - Hepatitis C virus has been proven to be the major cause of NANB hepatitis, cirrhosis and hepatocellular carcinoma worldwide. Based on the genome similarities between HCV and flavivirus or pestivirus, this agent has been included within the family Flaviviridae as a separate genus. Among the analogies between HCV and the other members of the same family there is the possibility of infecting blood cells. In particular, significant evidence obtained through studies performed in vivo and in vitro support the concept that HCV is not only a hepatotropic but also a lymphotropic virus. This suggests that, in addition to playing a role in inducing hepatic diseases (both of a non-tumoral and a neoplastic nature), HCV infection may also play a role in extrahepatic pathologies. The striking association observed between HCV infection and some autoimmune-lymphoproliferative disorders of either benign or neoplastic nature is consistent with this hypothesis. However, in analogy with what has been observed in the case of liver disease, the mechanisms involved in the pathogenesis of HCV related extra-hepatic manifestations have to be more deeply analysed and clarified. PMID- 8730475 TI - A laboratory and thrombokinetic study of HCV-associated thrombocytopenia: a direct role of HCV in bone marrow exhaustion? AB - OBJECTIVE: In order to investigate HCV associated thrombocytopenia, 6 patients suffering from this disease, in the absence of splenomegaly and other common causes of peripheral platelet destruction, underwent laboratory and scintigraphic tests. RESULTS: Thrombokinetic studies revealed a significant, nearly linear, delayed splenic accumulation with normal or low-normal values for the average platelet life span, low-normal recovery, and depressed platelet production. Megakariopoiesis was normal or slightly increased. HCV infection of the megakariocytes was found in two patients. Autoantibodies and liver disease were also investigated. CONCLUSIONS: A role of immunological mechanisms in HCV associated thrombocytopenia appears to be ruled out. The authors conclude that tests for HCV infection should be included in the evaluation of thrombocytopenia in adults and a possible direct involvement of HCV cannot be excluded. PMID- 8730476 TI - HCV and monoclonal gammopathies. AB - OBJECTIVE: To determine the prevalence of antibodies against HCV in monoclonal gammopathies with and without cryoglobulinemic activity. METHODS: 201 patients were divided into two groups: (I) 94 patients with monoclonal gammopathies with cryoglobulinemic activity, and (II) 107 with monoclonal gammopathies without cryoglobulinemic activity. Cryoglobulins were characterized by immunofixation; HCVAb were detected using second-generation ELISA and RIBA methods; in 38 cases the presence of HCV in peripheral blood mononuclear cells was evaluated by PCR. RESULTS: The HCVAb prevalence, as evaluated by RIBA, in Group I was 69.1% while in Group II it was only 14.9%. Histological and immunohistochemical study of the bone marrow in Group I patients frequently showed signs of nodular B-cell clonal expansion. CONCLUSIONS: Our data confirm the existence of a close correlation between HCV infection and the monoclonal gammopathies with cryoglobulinemic activity. HCV-positive cryoglobulinemic is characterized by self-limiting IgM monoclonal expansion associated with histological aspects of bone marrow lymphoid nodules that do not expand in the course of the disease like classic evolving lymphoproliferative processes. PMID- 8730477 TI - HCV and lymphoproliferative diseases. AB - METHODS: Genomic and replicative forms of HCV-RNA in B lymphocytes were detected by RT-PCR, and HCV genotyping was performed using universal and type-specific primers for the core region. Immunoglobulin gene rearrangement was detected by RT PCR. RESULTS: The presence of genomic and replicative forms of HCV-RNA in the 5'NC region was investigated on total RNA extracted from subpopulations of PBMC. The frequency of HCV-RNA was higher in the B lymphocytes than in other PBMC. In two patients a larger sized band was present in the B lymphocytes and PMN; this band could represent either another form of HCV-RNA or a cross-reaction between cellular RNA and HCV primers. HCV-RNA detected using primers for the core region was negative in the patients examined. Immunoglobulin monoclonal gene rearrangement was present on the cDNA in all of the HCV and type II cryoglobulinemia positive samples except two; in contrast, it was absent in the HCV positive and cryoglobulinemia negative samples. The analysis of immunoglobulin monoclonal gene rearrangement on DNA showed the presence of new positive samples among the HCV positive, type II cryoglobulinemia negative patients, who had been negative when PCR was performed on cDNA. Denaturing sequencing gel showed clearer results than agarose gel. CONCLUSIONS: The early detection of immunoglobulin monoclonal gene rearrangement and expression is very important because it could provide evidence of the possible lymphoproliferative evolution of HCV infection. In addition, these investigations together with PCR product sequencing could show us the steps in the clonal selection of B lymphocytes towards malignant transformation, in which HCV plays a direct and/or indirect role. PMID- 8730478 TI - "Single step" PCR with a sensitivity similar to nested PCR for the detection of hepatitis C virus RNA. AB - OBJECTIVE: Evaluation of the performance of different HCV PCR detection systems for HCV RNA: A nested PCR, considered the reference assay, was compared with two single-step methods (ss-PCR): the first is based on the detection of PCR products by liquid hybridization with a 32P end-labelled probe (isotopic ss-PCR), while the second assay is a colorimetric method (colorimetric ss-PCR) using microwell plate hybridization with a specific nucleic acid probe (Amplicor HCV PCR, Roche Diagnostics Systems). METHODS: Sera from 56 patients with suspected hepatitis C infection based on reactive serology or altered liver parameters, and sera from 15 blood donors were tested for HCV RNA: After RNA extraction, the synthesized HCV cDNA was amplified in parallel using isotopic ss-PCR, colorimetric ss-PCR and nested PCR. The products were detected by autoradiography, color development and ethidium bromide fluorescence, respectively. RESULTS: In order to assess the analytical sensitivity of ss-PCR versus that of nested of PCR, experiments included serial dilutions of positive control samples. Results showed that both methods had an extinction signal at the 1:512 dilution. A comparative analysis of 71 clinical sera samples was obtained using the three protocols and the results clearly documented 100% concordance. CONCLUSIONS: Single step PCR methods for HCV RNA have a sensitivity equal to that of nested PCR and appear more suitable for diagnostic applications. Ss-PCR is safer than nested PCR in terms of both specificity and contamination problems. In particular, the Roche Amplicor HCV PCR assay minimizes sample exposure and management problems. PMID- 8730479 TI - Viruses and cryoglobulinemia. AB - Circulating cryoglobulins have been reported in association with several acute and chronic viral diseases. In all of the reported cases, the viruses involved have been hepatotropic, lymphocytotropic or both. Among the hypotheses concerning the causes of cryoglobulinemia, two possible pathways have been more frequently debated: an impairment of the macrophagic system of the liver, with the consequent impairment of the clearance of gut antigens and immunoglobulins, as the first ring of a chain of events including the activation of the B cell compartment with increased production (and decreased clearance) of cryoglobulins; and a low-grade malignant lymphomatous process involving rheumatoid factor producing clones. Recent evidence of a close association between hepatitis C virus (HCV) and "essential" mixed cryoglobulinemia has focused the attention of several researchers on the mechanisms by which the virus is capable of causing cryoglobulin synthesis. The open questions include: (1) Why do only a minority of chronically HCV-infected people (mainly "sporadically" infected elderly women) develop a cryoglobulinemic syndrome? (2) What kind of mechanisms can up- or down regulate cryoglobulin production? (3) Are immunoregulatory mechanisms involved? (4) Is there a connection between HCV infection and the low-grade malignant lymphoma hypothesis identifying cryoglobulinemias as the consequence of the slow proliferation of CD5+ B cells? and (5) Are particular HCV genotypes specifically involved in causing cryoglobulinemias? Several of these questions still remain unanswered. HCV has been detected in the peripheral blood mononuclear cells of both cryoglobulinemic and non-cryoglobulinemic infected subjects; on the other hand, the detection of viral RNA in the bone marrow cells of virtually all cryoglobulinemic patients suggests that this might be related to the pathogenesis of the disease. PMID- 8730480 TI - Immunochemical characterization of the cryoglobulins: pathophysiologic implications. AB - Immunochemical study of the cryoglobulins has produced a classification of three main types which present clinical correlates. With the current standard immunofixation technique the vast majority of cryoglobulins are presently classified as type II. High-resolution immunoelectrophoretic techniques (immunoblotting and two-dimensional polyacrylamide gel electrophoresis) have helped to identify new sub-types whose clinical relevance is as yet undetermined. Nevertheless, these sensitive techniques could contribute to elucidate the various stages in the natural history of HCV-related cryoglobulins, from possible earlier HCV-antigen-driven polyclonal B cell proliferation, to autonomous monoclonal benign, to malignant B cell proliferation. PMID- 8730481 TI - Characterization of cryoglobulins: some remarks on methodology. AB - OBJECTIVE: The experience of our group in the characterization of the cryoglobulins during the last 8 years is reported, and the possible advantages of immunofixation versus immunoblotting are discussed. METHODS: 25 out of 171 cryoprecipitates studied by immunofixation were also examined by immunoblotting, a technique which has been suggested to offer advantages in analysing monoclonal components. RESULTS: We confirm that immunofixation may be the technique of choice given its easier execution and interpretation, better standardization, and lower overall cost. PMID- 8730482 TI - HCV genotypes and cryoglobulinemia. AB - OBJECTIVE: To evaluate the HCV genotype distribution in subjects affected by cryoglobulinemia in order to verify its possible role in the pathogenesis of the disease and to provide the clinician with a useful datum for therapy. METHODS: Nested PCR with universal and type-specific primers was used for the genotyping. RESULTS: Genotype I (1a) was never present in cryoglobulinemia, while it was present in 7 (4.3%) patients with chronic hepatopathy and in 4 (10.8%) asymptomatic patients. Type II (1b) was present in 28 (58.3%) and in 8 (47.1%) cryoglobulinemic patients with and without hepatopathy, respectively, in 106 (64.6%) patients with chronic hepatitis; in one patient with acute hepatitis; and in 14 (37.9%) asymptomatic patients. Type III (2a) was present in 2 (4.2%) and 2 (11.8%) cryoglobulinemic patients with and without hepatopathy, respectively; in 1 (0.6%) patient with chronic hepatopathy; and in 2 (5.4%) asymptomatic subjects. Type IV (2b) was present in 1 (2.1%) and in 2 (11.8%) cryoglobulinemic patients with and without hepatopathy, respectively; in 5 (3%) patients with chronic hepatopathy; and in 1 (2.7%) asymptomatic subject. Coinfections were present in 42 cases: 6 (12.5%) cryoglobulinemia with hepatopathy, 4 (23.5%) cryoglobulinemia without hepatopathy, 25 (15.3%) chronic hepatopathy, and in 7 (18.9%) asymptomatic subjects. For 41 (15.4%) strains typing was not possible. Eight of the "untypable" strains and 3 strains from patients with coinfection proved to belong to a new genotype. CONCLUSIONS: Genotype II (1b) was the most frequent in patients with and without cryoglobulinemia; genotype I (1a) was absent in all 65 patients with cryoglobulinemia, in whom, however, as in the subjects without cryoglobulinemia, all the other genotypes could be found. An interferon-resistant genotype characterized by an elevated homology with Simmonds' type 2c (rare genotype) was present. PMID- 8730483 TI - Significance of hepatitis C virus genotypes and viral load in patients with "essential" mixed cryoglobulinemia. AB - Several studies have established a strong association between hepatitis C virus (HCV) infection and essential mixed cryoglobulinemia (EMC). However, the mechanisms by which HCV infection may result in cryoglobulinemia in some patients but not in others remain unknown. In this paper we shall summarize some of the work done in our laboratories on certain aspects of HCV in patients with EMC. PMID- 8730484 TI - HCV genotypes in bone marrow and peripheral blood mononuclear cells of patients with mixed cryoglobulinemia. AB - OBJECTIVE: The association of the hepatitis C virus (HCV) with the cryoglobulinemic syndrome is well known, but its pathogenetic mechanism still remains to be clarified. HCV-RNA has been found in the peripheral blood mononuclear cells (PBMC) of infected subjects. We investigated the presence of the HCV genome in bone marrow cells (BMC), and the distribution of different HCV genotypes in individuals with mixed cryoglobulinemia (MC) and in noncryoglobulinemic controls. METHODS: 15 anti-HCV positive subjects with MC, 7 non-cryoglobulinemic patients with type C chronic active hepatitis (CAH) and 2 anti-HCV negative controls were studied. HCV-RNA was detected by nested PCR of the highly conserved 5'-NCR sequence. HCV typing was carried out by means of the hybridization of the same amplified region with specific probes. RESULTS: HCV-RNA was present in the PBMC of a large proportion of the MC patients and controls without any significant differences. On the contrary, HCV-RNA was present in the bone marrow cells of all the patients with MC and in 43% of the CAH controls. The HCV 1b and 2a genotypes seem to be the most prevalent among MC patients. Nevertheless, the patients with type II MC had a very high prevalence of the 2a genotype (77%). CONCLUSION: The results suggest that the presence of HCV-RNA in bone marrow cells may be correlated to the pathogenetic mechanism of MC. Other studies are needed to confirm the frequent association of HCV genotype 2 with MC. PMID- 8730485 TI - HLA expression in type II mixed cryoglobulinemia and chronic hepatitis C virus. AB - OBJECTIVE: Hepatitis C virus infection is closely associated not only with hepatic damage, but also with mixed cryoglobulinemia (MC) and other autoimmune and lymphoproliferative disorders. Because HCV is both hepatotropic and lymphotropic, the aim of this study was to investigate whether the genetic background may influence the clinical pattern seen in different patients. METHOD: Two groups of patients with HCV infection were studied: 16 with type II MC and 18 with chronic active hepatitis (CAH). 120 bone marrow donors were considered as the control group. In all patients HLA-A-B-C antigens were evaluated using the microlymphocytoxicity technique, and HLA-DR by the PCR-SSP method. RESULTS: The frequency of the HLA antigens expressed was not precisely defined in the two groups. However, the HLA-B51 and B35 antigens, which are often correlated with autoimmune disorders, were highly expressed in the MC patients (31.2%) compared to the controls (6.9%) and to the CAH group (11%). Moreover, HLA-A9 with its split A24 were present in 50% of the MC patients. More interesting was the expression of the HLA-DR7 antigen, which was found only in the CAH group, suggesting that it may influence the specific liver involvement in HCV infections. CONCLUSION: These findings indicate that the HLA system may play an important role in the clinical manifestations of HCV infection. PMID- 8730486 TI - Humoral immune response and natural killer activity in patients with mixed cryoglobulinemia. AB - OBJECTIVE: Based on serological and molecular evidence of hepatitis C virus (HCV) infection in a significant proportion of patients with mixed cryoglobulinemia (MC), a direct association between HCV and MC has been suggested. The goal of the present study was to investigate the role played by HCV and by the immune response to the virus in the pathogenesis of mixed cryoglobulinemia. METHODS: A competitive reverse transcription polymerase chain reaction was employed to evaluate the concentrations of specific HCV RNA sequences in different clinical specimens (plasma, sera, cryoprecipitates, bone marrow and peripheral blood cells). Using recombinant and synthetic peptides covering the HCV core, envelope 1 (E1) and nonstructural regions 4 (NS4) and 5 (NS5), the humoral immune response in a group of MC patients was assessed with an enzyme-linked immunosorbent assay. Natural killer (NK) cell activity was estimated using a 4 hr 51 Cr release assay. RESULTS: Quantitation of the RNA molecules in the biological samples confirmed an increased virion concentration in cryoprecipitates from 13/15 patients with mixed cryoglobulinemia. Analysis of the humoral immune response against the synthetic peptides suggested a distinct response to HCV antigens in MC patients when compared to patients with HCV infection but without serological evidence of cryoglobulinemia. Unstimulated NK cell functioning was below the normal range in all patients tested. However, peripheral blood mononuclear cells showed no enhancement of NK activity by the interferon inducer polyinosinic acid:polycytidilic acid. Enhancement by interferon-alpha was normal, suggesting an impairment in interferon production. CONCLUSION: The quantitative data are in line with the hypothesis of a direct or indirect role of HCV in mixed cryoglobulinemia. The abnormal immune response could be involved in the onset and persistence of HCV infection, and possibly in the appearance of cryoglobulinemia. PMID- 8730487 TI - The association of hepatitis C virus infection with monoclonal rheumatoid factors bearing the WA cross-idiotype: implications for the etiopathogenesis and therapy of mixed cryoglobulinemia. AB - OBJECTIVE: To determine the prevalence of monoclonal rheumatoid factors (mRF) bearing the WA cross-idiotype (WA XId) in hepatitis C virus (HCV) positive type II mixed cryoglobulins, to review recent studies on the role of HCV in the cutaneous vasculitis lesions in patients with type II cryoglobulinemia and to discuss the implication of these studies for the etiopathogenesis and therapy of the disease. METHODS: Thirty type II cryoglobulins were tested for WA and PO XId and for HCV RNA: RESULTS: WA mRF were strongly, although not exclusively, associated with HCV in type II mixed cryoglobulinemia. CONCLUSION: These and other recent studies from our laboratory suggest that chronic HCV infection may be the stimulus for the production of WA mRF and that HCV may be directly involved in the pathogenesis of the cutaneous vasculitis in patients with type II cryoglobulinemia. The association of HCV infection with the disease provides a rationale for anti-viral therapy and for monitoring therapy by measuring the HCV level in both blood and liver. PMID- 8730488 TI - Cross-reactive idiotypes on cryoprecipitating monoclonal IgMk rheumatoid factors. AB - Shared idiotype specificities (CRIs) among monoclonal IgM rheumatoid factors from patients with essential mixed cryoglobulinemia have been demonstrated with the use of polyclonal antisera and have been confirmed, more recently, using monoclonal antibodies. In this paper we will summarize some of the work that has been performed in our laboratory using anti-idiotypic monoclonal antibodies. Such reagents allowed us to detect CRIs on cryoprecipitable rheumatoid factors, to detect idiotype-positive cells in bone marrow and peripheral blood, and to identify glomerular immune deposits. PMID- 8730489 TI - Relationship between rheumatoid factor and the immune response against hepatitis C virus in essential mixed cryoglobulinemia. AB - OBJECTIVE: The authors examine the relationship between the presence of hepatitis C virus (HCV) and anti-HCV antibodies, rheumatoid factor (RF) activity, the level of complement, and the cryocrit in the sera of patients affected by essential mixed cryoglobulinemia (EMC). In addition, the authors evaluate the circulating B CD5 positive lymphocytes, believed to be RF producers in the blood of EMC patients. METHODS: Clinical and laboratory data on 219 cases of EMC were collected from five centers of the GISC (Italian Group for the Study of Cryoglobulinemias). Statistical analyses of these data were carried out with the aim of evidentiating significant relationships, in order to shed light on the mechanism of cryoprecipitation. RESULTS: The cryocrit was higher in anti-HCV negative and in HCV-RNA positive sera. The titers of RF activity were significantly higher in anti-HCV negative sera. A linear correlation between RF activity and the cryocrit was observed for HCV-RNA positive cases (r = 0.416), this correlation being highest in the patients who were HCV-RNA positive and anti HCV negative (0.709). Type III EMC were more reactive than type II against the non-structural antigens of HCV, particularly 5-1-1 and C-22. There were more circulating B lymphocytes sharing membrane antigen CD5 in EMC than in the blood of normals or patients with HCV-related chronic hepatitis. Levels of circulating B-CD5 correlated with serum RF activity (r = 0.677), especially in sera from anti C-100 negative patients (p = 0.991). CONCLUSION: The data strongly suggest that the level of the cryoprecipitate is a function of both the HCV-RNA in the serum and the RF activity. Antibody specificity against HCV probably influences the density of the cryoprecipitate. PMID- 8730490 TI - Lymph node angiogenesis in lymphoproliferative disorders: type II Mixed cryoglobulinemia as a model. AB - OBJECTIVES: Lymph node biopsies from patients with mixed cryoglobulinemia (MC) were investigated for: a) the presence of the hepatitis C virus (HCV) peptides C22 and C100; b) the degree of angiogenesis and of inflammatory infiltrate (macrophages and neutrophils); c) the distribution of laminin and type IV collagen, the main components of the subendothelial basement membrane; and d) the ultrastructural characteristics of the microvessels. METHODS: Cryostat sections were stained by immunoperoxidase to highlight HCV peptides, endothelial cells, their basement membrane components, and inflammatory cells. Planimetric methods were applied to count the microvessels and inflammatory cells, and to evaluate the distribution pattern of the basement membrane components. Microvessels were also analysed at the ultrastructural level. RESULTS: HCV+ lymph nodes showed higher microvessel and inflammatory cell counts than the HCV- lymph nodes. Their laminin and type IV collagen distribution and ultrastructural findings also indicated immature capillaries. CONCLUSIONS: HCV could be introduced into the lymph nodes by infected mononuclear cells, and trigger inflammation and angiogenesis, thus contributing to the inflammatory process. Angiogenesis, in turn, could facilitate HCV dissemination and persistence in MC. PMID- 8730491 TI - HCV-RNA detection using different PCR methods in sera, cryoglobulins and peripheral blood mononuclear cells of patients with mixed cryoglobulinemia. AB - OBJECTIVE: The hepatitis C virus (HCV) is frequently associated with mixed cryoglobulinemia (MC), and a number of authors have reported the presence of anti HCV antibodies and HCV-RNA in the blood of MC patients. The presence of the HCV genome in the blood cells of individuals infected by HCV may correlate with the etiopathology of MC. We investigated the presence of HCV-related sequences in the sera, cryoglobulins and peripheral blood mononuclear cells (PBMC) of patients with MC and of individuals with type C chronic active hepatitis (CAH). METHODS: 39 patients with MC, 11 non-cryoglobulinemic HCV-positive individuals with CAH, and 2 anti-HCV negative controls were included in the study. The presence of HCV RNA was detected by nested RT-PCR and by a commercial kit. The PCR was performed by amplifying the 5'-non coding region (5'-NCR) of HCV. RESULTS: HCV-RNA was detected in the sera and cryoglobulins of about 90% of the patients; the commercial kit showed a higher sensitivity than nested PCR. One MC patient showed HCV-RNA only in the cryoglobulins. HCV-RNA was present in the PBMC of 14 of the 20 (70%) MC patients analyzed. No differences in serum and PBMC HCV-RNA positivity were found between MC patients and controls. CONCLUSION: Our results confirm the spread of HCV infection among patients with MC. HCV-RNA is present in the serum, cryoglobulins and PBMC of a large proportion of MC patients. The prevalence of HCV-RNA in the PBMC of MC patients and controls did not differ significantly; this may suggest a tropism of HCV for PBMC regardless of the presence of cryoglobulinemia. PMID- 8730492 TI - Classification of the cryoglobulinemic syndrome. AB - As cryoprecipitation is a laboratory artifact, it is necessary to identify those patients with mixed cryoglobulinemia who actually have the cryoglobulinemic syndrome. In certain cases, although by no means frequently, mixed cryoglobulins are associated with a vasculitis that produces a spectrum of manifestations resembling those of experimental serum sickness. Therefore, we propose a set of criteria to be used as the starting point for establishing a definitive diagnosis. The next step in the classification of cryoglobulinemic syndromes is to define the primary or secondary nature of the disease process and the mono- or polyclonal nature of the rheumatoid factor. Finally, the extent of the vasculitis must be assessed by screening for liver and renal involvement and peripheral neuropathy. Despite the high prevalence of HCV markers in both the primary and secondary forms, we believe our classification is useful for characterizing groups of patients with different diseases and different prognoses. PMID- 8730493 TI - The natural history of cryoglobulinemia: symptoms at onset and during follow-up. A report by the Italian Group for the Study of Cryoglobulinemias (GISC) AB - OBJECTIVE: The cryoglobulinemic syndrome (CS) may be associated with other diseases; when it is not, it is termed essential. Recently the natural history of this disease has been re-evaluated on the basis of its close association with HCV markers. In this context, the GISC has studied a large series of patients from several Italian centres. METHODS: In a multicentric retrospective study of 913 cases, we evaluated the clinical and laboratory signs at onset in patients with essential mixed cryoglobulinemias (EMC) (654 cases) or secondary cryoglobulinemias (SC) (259 cases) and sought to define possible diagnostic criteria typical of the different CS. We also carried out a retrospective 5-year cohort study on 192 patients selected randomly from the 913 cases described above. In particular, we examined the correlation between the presence/absence of concomitant diseases and the presence of HCV markers on the clinical evolution and survival of the patients. RESULTS: Purpura, rheumatoid factors (RF), significant C4 consumption, and Brouet's classification type II were more frequent in EMC. Purpura, Meltzer's triad, and Raynaud's phenomenon improved, while renal involvement tended to worsen over time. During the follow-up we did not note a significant change in clinical staging in the patients with Brouet's type II cryoglobulins, in the patients with HCV-related markers, or in the overall series. The most frequent causes of death in 34 patients were liver, cardiovascular, renal, and lymphoproliferative diseases. Lymphomas were diagnosed in 11 patients during follow-up, with particular frequency in the HCV-marker positive patients. CONCLUSION: Specific clinical and laboratory features typical of the different CS subgroups could not be identified on the basis of these data. Our follow-up data seem, however, to confirm a role for HCV in the mixed cryoglobulinemias. PMID- 8730494 TI - Etiopathogenetic role of hepatitis C virus in mixed cryoglobulinemia, chronic liver diseases and lymphomas. AB - Hepatitis C virus (HCV) infection has been found in the majority of patients with mixed cryoglobulinemia (MC) in studies conducted in different countries. In our series of 110 MC patients the frequency of HCV markers was significantly high (91%) compared with other rheumatic diseases (6.4%) and with healthy Italian controls (1.2%). Moreover, HCV RNA was detected in 81% of the peripheral lymphocytes from MC patients. Comparable percentages of HCV infection were detectable in other disorders, i.e. porphyria cutanea tarda (77%) and autoimmune hepatitis type 1 (77%). The HCV infection of peripheral lymphocytes suggests that this virus could be the triggering factor for the lymphoproliferation underlying MC. In a number of patients with MC the evolution from a benign lymphoproliferation to frank B-cell lymphoma was observed. In these subjects HCV RNA in the sera and in fresh and cultured peripheral lymphocytes was constantly detected. The same phenomenon has been observed in patients with long-lasting type C chronic hepatitis. Interestingly, HCV infection has also been recorded in 32% of idiopathic B-cell non-Hodgkin's lymphomas. Taken together, the above findings suggest that HCV can cause benign B-cell proliferation with the consequent production of various autoantibodies, including rheumatoid factor and mixed cryoglobulins. These serological abnormalities characterise different clinical disorders, including the appearance of lymphoma in a not negligible number of individuals. PMID- 8730495 TI - Lymphoproliferation in type II mixed cryoglobulinemia. AB - OBJECTIVE: The role of the hepatitis C virus (HCV) in the etiology of type II mixed cryoglobulinemia (MC) has been well established, but the pathogenetical relationships among the virus, the immune system, the natural history of MC, and lymphoproliferation in the bone marrow and liver need to be further elucidated. METHODS: Eighty-two patients with HCV positive type II MC and 20 subjects with chronic hepatitis C without MC were studied: bone marrow and liver specimens were examined by routine histology and immunohistochemistry, particularly focusing on parameters related to disease behaviour, such as the expression of the bcl-2 oncogene product and the proliferation-associated Ki67 antigen. RESULTS: In most MC patients there were lymphoid infiltrates within the bone marrow showing a monomorphic cytology, frequent immunoglobulin light chain monotypic restriction, expression of the anti-apoptotic bcl-2 oncogene product, and a low proliferative capacity (Ki-67 < 3%). On the other hand, in all non-cryoglobulinemic patients a bone marrow picture of reactive lymphoplasmacytosis was found. In both MC and chronic hepatitis patients, the liver biopsy showed portal infiltrates consisting of T-cells, associated with a significant B-cell component; the latter was particularly abundant in MC, where it was frequently arranged in pseudo follicles. The B-cell component expressed the bcl-2 oncogene product and CD5 antigen, thus suggesting that the immune system is actively involved in the production of liver damage both in MC and non-cryoglobulinemic patients. It is worth noting that in MC patients (but not in the non-cryoglobulinemic patients) these CD5+/bcl-2+ B-cells frequently also exhibited a monotypic restriction bearing an IgM kappa. CONCLUSION: Our findings in these liver and bone marrow studies further support the role of a lymphoproliferative disorder in the pathology of type II MC: the B cells involved accumulate due to the inhibition of apoptosis, and their low proliferative index justifies the indolent course of the disease. HCV probably interacts with these B-cells, facilitating their clonal expansion. PMID- 8730496 TI - Cryoglobulinemia in transfusion-dependent thalassemia major. AB - OBJECTIVE: The aim of this study was to determine the frequency of cryoglobulinemia and associated symptoms in transfusion-dependent thalassemia patients at high risk for HCV infection. METHODS: A controlled epidemiological study was used to evaluate the prevalence of clinical, biochemical and immunological abnormalities in a group of 264 HCV-positive and 106 HCV-negative transfusion-dependent thalassemia patients. Haematologic and hepatic function tests were performed according to standard methods. HCV-RNA was detected by PCR analysis. RESULTS: The significant presence of cryoglobulinemia and associated symptoms (purpura, vasculitis, arthritis, asthenia, proteinuria), serum autoantibodies (SMA, anti-GOR, ANA, LKM), low complement and rheumatoid factor were found in HCV-positive compared with HCV-negative patients. CONCLUSIONS: This study demonstrates the role of HCV in inducing cryoglobulinemia and immunological disorders in transfusion-dependent thalassemia patients. HCV infection and associated immune abnormalities are a new clinical aspect of, and deserve particular attention due to their high frequency in, transfusion-dependent thalassemia patients. PMID- 8730497 TI - Cryoglobulinemia in HCVAb+ patients on chronic hemodialysis. AB - OBJECTIVE: Striking evidence of HCV infection has been found in mixed cryoglobulinemia (MC) and HCV has been hypothesized to be the causative agent of this disease. To assess the association of C virus infection and cryoglobulinemia we studied cryoglobulin levels in 66 patients on maintenance hemodyalisis who were selected on the basis of HCVAb positivity and not because they were affected by liver disease. The control group was made up of 45 patients also on hemodyalisis but without HCV infection. RESULTS: Circulating cryoglobulins were found in 34 (52%) of 66 HCV+ patients: the cryocrit was < 1% in 20, 1 to 5% in 12, and > 5% in 2 patients. The cryoglobulins were classified by immunofixation as type II in 8 and type III in 8 others; identification was not possible in 18 cases. In the HCVAb- control group untypable cryoglobulins were detected in 9% of the patients at < 1% by volume. No correlation was found between these data and the liver disease detected by biohumoral tests. CONCLUSIONS: Our data confirm the close link between HCV infection and cryoglobulins; the prevalence of circulating cryoglobulins in uremic HCVAb+ patients is very close to that found in HCV related liver disease. PMID- 8730498 TI - Disappearance of cryoglobulins and remission of symptoms in a patient with HCV associated type II mixed cryoglobulinemia after HIV-1 infection. AB - OBJECTIVE: We report the case of a woman with long-lasting mixed cryoglobulinemic syndrome, who experienced clinical and laboratory remission of her cryoglobulinemia after becoming infected with human immuno-deficiency virus and developing HIV-1 induced immunosuppression. METHODS: Serum cryoglobulin concentrations and the CD4+ cell count were monitored every three months. RESULTS: After the diagnosis of HIV-1 infection, the immunological status of the patient was constantly depressed (CD4+ cell count dropping from 337/microL in January 1991 to 21/microL in June 1994). Serum cryoglobulins were persistently absent over 43 months of follow-up, despite the presence of HCV-RNA. CONCLUSION: In this case, HIV-1 induced immunodeficiency seems to be responsible for the remission of the cryoglobulin syndrome and the disappearance of serum cryoglobulins. These findings indicate that CD4+ T lymphocytes may play a role in regulating the activity of cryoprecipitating rheumatoid factor secreting B cell clones. PMID- 8730499 TI - Hepatitis C antibody and cryoglobulins in patients with cirrhosis and hepatocellular carcinoma. AB - OBJECTIVE: To assess: (1) the role of hepatitis C virus (HCV), hepatitis B virus (HBV) and alcoholism as risk factors for hepatocarcinoma (HCC) in patients with liver cirrhosis, and (2) the presence of cryoglobulins in HCV + patients with and without HCC. PATIENTS: 82 cirrhotic patients, 41 with and 41 without HCC, who were admitted consecutively to our General Medicine Division from January 1992 to June 1994, were studied. RESULTS: The prevalence of HBV markers, anti-HCV and alcoholism in patients with liver cirrhosis and HCC was 39%, 63.4% and 24%, and in cirrhotics without HCC it was 31.7%, 51.2% and 63.4%, respectively. Cryoglobulins were present in 87.5% of the patients with HCC (78.5% anti-HCV +) and in 57.8% of the patients without HCC (81.8% anti-HCV +). The cryoglobulins, as characterized in 11 cases, were type III in nine cases and type II in two. CONCLUSIONS: Patients with cirrhosis, especially when associated with HCV and HBV infection, are at high risk for HCC and therefore require careful follow-up. Moreover, a strong association between HCV and the cryoglobulins in cirrhotics with and without HCC was evident, thus supporting the possible role of this virus in stimulating lymphocytes to produce cryoglobulins. PMID- 8730500 TI - Anti-GOR antibodies, HCV, liver disease and cryoglobulinemia. AB - OBJECTIVE: We investigated the presence of anti-GOR antibodies in patients with essential mixed cryoglobulinemia, since both autoimmune pathogenetic processes and a high prevalence of HCV infection are present in this syndrome. METHODS: We compared these cases to patients with HCV-related chronic active hepatitis or alcoholism, and to ex-blood donors. A total of 60 patients with biopsy-proven chronic liver disease were studied. RESULTS: HCV related markers, cryoglobulins, anti-GOR antibodies and ANA were detected in all of the groups. CONCLUSION: Our data would appear to indicate that anti-GOR are related to the presence of HCV chronic hepatitis and not to cryoglobulinemia or chronic liver damage. PMID- 8730501 TI - Alpha interferon treatment in chronic hepatitis C. AB - INTRODUCTION: Alpha interferon (alpha IFN) treatment normalizes serum ALT levels in at least half of all patients affected by chronic hepatitis C, but a reactivation of the disease is frequently observed after the end of therapy. Different regimens of alpha IFN therapy have been proposed but the optimal schedule is still controversial. Recently at least 6 different types of HCV have been identified and the HCV genotype has been proposed as an important factor influencing the response to alpha IFN therapy. OBJECTIVE: The aim of this study was to evaluate the efficacy of different regimens of alpha IFN in chronic hepatitis C, and to study the relationship between the response to treatment and HCV genotypes. METHODS: 160 consecutive patients affected by biopsy-proven chronic hepatitis C were randomly treated with different doses of lymphoblastoid alpha IFN [adjusted to the body surface area (m2)] and different durations of therapy, as follows: 2 MU/m2/t.i.w. for 6 or 12 months or 4 MU/m2/t.i.w. for 6 or 12 months. Biochemical and virological responses were studied: ALT levels were monitored monthly during and for at least 6 months after the end of treatment, and serum HCV RNA was assessed before and at the end of therapy, using nested RT PCR. Biochemical responses were defined in advance as follows: non-response as maintenance of abnormal ALT levels during treatment; complete response as the normalization of ALT by the 4th month and lasting until the end of treatment; sustained response as a complete response lasting for at least 6 months after the end of therapy. The clearance of serum HCV RNA at the end of therapy was considered a virological response. In pre-treatment sera stored at-80 degrees, HCV genotyping was performed according to the method of Okamoto. The Chi square test and multiple stepwise logistic regression were used for the statistical analysis. RESULTS: A sustained biochemical response was significantly more frequent in patients treated for 12 than in patients treated for 6 months, independently of the dosage (45% vs 24% in patients treated with 2 MU/m2/tiw, and 55.5% vs 30% in those treated with 4 MU/m2/tiw). The distribution of HCV genotypes (according to the classification of Okamoto) was 9.8% type I, 45.5% type II, 37.1% type III and 7.6% type V. Both the biochemical and virological responses were significantly correlated to the HCV genotype, being significantly more frequent in patients infected with type III or V (71-60% biochemical and 48 50% virological response, respectively) than in patients with type I or II (15% biochemical and 18-21% virological response, respectively). CONCLUSIONS: 12 months of alpha IFN treatment seemed to be significantly more efficacious than 6 months of therapy, and a significant relationship between the HCV genotype and the biochemical and virological response to alpha IFN was found. PMID- 8730502 TI - Interferon in the treatment of mixed cryoglobulinemia. AB - OBJECTIVE: To assess the efficacy of alfa-IFN in HCV-associated type II cryoglobulinemia. METHODS: An open trial was carried out on 24 patients with HCV associated type II mixed cryoglobulinemia using recombinant alfa-IFN (3 MU three times weekly for 12 months). The patients were followed for at least 18 months and visceral involvement was evaluated before and after IFN using a scoring system. RESULTS: Alfa-IFN treatment had a marked effect on skin and liver involvement, while a moderate response was obtained in relation to the nephropathy and peripheral neuropathy. A significant lowering of the cryocrit and an increase in serum C4 were observed. Eleven out of the 16 patients who responded favourably relapsed within six months. In 3 patients, all complete responders, HCV-RNA became undetectable at the end of treatment. CONCLUSION: Alfa IFN may be regarded as the treatment of choice in HCV-associated type II MC, but further studies are required to clarify the factors associated with the lack of response or the relapses seen in some patients. PMID- 8730503 TI - Effects of two different alpha-interferon regimens on clinical and virological findings in mixed cryoglobulinemia. AB - OBJECTIVE: As previous studies have shown a good response of mixed cryoglobulinemia (MC) to alpha-interferon (IFN) therapy, we investigated the efficacy and tolerability of two IFN regimens in a group of 36 patients affected by MC. METHODS: The patients, diagnosed on the basis of standard clinical and laboratory criteria, were randomly divided into 2 groups: group A (18 cases) received alpha 2b-IFN 3 M.U. thrice a week for six months, while group B (18 cases) received alpha 2b-IFN thrice a week for 1 year. The patients were followed for six months after the end of therapy. Liver function tests, cryoglobulin determinations and a clinical examination were performed each month. HCV serology and HCV-RNA detection by PCR were performed before therapy and at the end of the follow-up period. RESULTS: The two study groups were comparable in age, male/female ratio, purpura score, cryoglobulin level, mean ALT serum activity and liver histology. 32 patients (89%) were positive for anti-HCV antibodies and 29 (81%) for HCV-RNA. During therapy all patients showed a significant (p < 0.001) decrease in their cryoglobulin level as well as improvement (p < 0.05) in their purpura score. In group A, five patients (28%) showed normalized ALT, but three later relapsed. In group B seven patients (39%) responded to treatment but three relapsed after suspension of the drug. Two patients from group B developed severe side effects (hypothyroidism and depression) and therapy was discontinued after 9 and 11 months, respectively. In all the non-responders and relapsed patients, purpura, ALT, and cryoglobulins rose to pre-treatment levels within a few months. At the end of follow-up, two patients from group A (11%) and four in group B (22%) had achieved complete remission. CONCLUSION: This study indicates that IFN is useful in MC and that viral replication can be considered the target of the therapy. Despite the absence of a statistical difference in the response rate between the two regimens (due to the low number of subjects), the one-year therapy course seemed to show a better efficacy, although associated with higher toxicity. PMID- 8730504 TI - GISC protocol experience in the treatment of essential mixed cryoglobulinaemia. AB - OBJECTIVE: We compared the efficacy of interferon and deflazacort in the treatment of the cryoglobulinaemic syndrome and assessed the usefulness of adding a low antigen diet to drug therapy. METHODS: We studied 63 patients randomly allocated to different groups who underwent clinical and laboratory examinations every two months and who received treatment for 12 months or until a significant clinical event appeared. RESULTS: Five of 28 patients treated with interferon showed clinical improvement whereas 4 worsened and 7 suffered untoward side effects; seven of 28 patients treated with deflazacort improved, 4 worsened and 4 suffered drug toxicity. Twenty-nine patients were assigned to combined low antigen diet and therapy, among whom 7 did not follow the diet, 5 improved and 2 worsened. Among the 34 patients who were on an unrestrained diet, 5 improved and 7 worsened. None of the treatments proved superior to the others. CONCLUSION: Our results do not confirm the suggestion that interferon should be the primary therapy in the treatment of the cryoglobulinaemic syndrome, and the usefulness of a low antigen diet seems minimal. PMID- 8730505 TI - Guidelines for the management of essential mixed cryoglobulinemia. AB - The principal therapeutic procedures and when they are clinically indicated in the management of essential mixed cryoglobulinemia (EMC) have been the subject of much debate. This paper reviews current knowledge and our experience in the treatment of this complex disease. It is generally agreed that patients with purpura, the primary symptom of EMC, should avoid long periods of sitting or standing in the same position. Non-steroidal antiinflammatory drugs can be used for the management of arthralgias and/ or arthritis. Low dose steroids (0.5-0.3 mg/kg/die) are usually sufficient to control the purpura, arthralgias, arthritis and weakness, while larger doses (0.5-1.5 mg/kg/die) are needed to treat the renal involvement, peripheral neuropathy and serositis. Since the discovery of the association between EMC and viral infections, the appropriateness of cytotoxic drugs has been re-evaluated and they are no longer used. With the low antigen content diet, a regimen designed to restore a saturated mononuclear phagocytic system, good results have been obtained in the treatment of purpura, arthralgias, weakness and peripheral neuropathy. Furthermore, this dietary regimen may play a steroid sparing role. Plasma exchange is widely used in the management of severe renal involvement, hyperviscosity syndrome, sensory motor neuropathy and liver involvement in EMC. PMID- 8730506 TI - Colchicine in the treatment of mixed cryoglobulinemia. AB - OBJECTIVE: The best treatment for cryoglobulinemic syndrome (CS) is still an unsolved problem. Recently colchicine has been successfully used to treat vasculitides and other immune-mediated diseases. Therefore, we undertook to treat 17 CS patients with colchicine (1 mg/day for 6-48 months), 8 of them with essential mixed cryoglobulinemia (EMC) and 9 with CS secondary to liver disease. METHODS: In all patients the clinical and laboratory features were evaluated at the beginning of the study and during the first 6-12 months; 10 cases were followed for a longer period (18-48 months). RESULTS: During the first period symptoms improved as follows: purpura in 15 of 17 patients, weakness in 9 of 14 and leg ulcers in 3 of 5. Hepatic and renal function tests, hypocomplementemia, rheumatoid factor (RF) titres and the cryocrit also improved. Prolonged follow-up showed a relapse in the different variables, although they remained at better levels than at the beginning. Only the cryocrit showed a further reduction. CONCLUSION: Though this was a preliminary open study it shows that colchicine is an efficient treatment in CS and suggests that a controlled clinical study should be performed to assess its real value. PMID- 8730507 TI - A pilot study on the effectiveness of cyclosporine in type II mixed cryoglobulinemia. AB - OBJECTIVE: To evaluate the effect of cyclosporine in the treatment of type II mixed cryoglobulinemia, after the failure of more conventional therapies. METHODS: Two patients with type II mixed cryoglobulinemia associated with chronic HCV infection, purpura, liver disease, and sensitive/motor neuropathy were treated with cyclosporine (2.5 mg/Kg/b.w.), after their failure to respond to treatment with corticosteroid, immunosuppressive drugs, interferon, and plasmapheresis. RESULTS: In both patients an improvement in the clinical manifestations (purpura and peripheral neuropathy), laboratory results (serum transaminases and cryocrit), and liver histology was seen, as well as the disappearance of bone marrow B-cell lymphoproliferation. CONCLUSION: Cyclosporine may be useful in the treatment of type II mixed cryoglobulinemia with prominent autoimmune clinical manifestations, although further studies are needed to better define the selection of patients. PMID- 8730508 TI - Hepatitis C virus: from biology to clinical practice. PMID- 8730509 TI - SH2 domain protein interaction and possibilities for pharmacological intervention. AB - SH2 domain containing proteins play a key role in the process of intracellular transmission of signalling events initiated at the cell surface. As a pre requisite in the fulfillment of this function, these proteins bind to a variety of phospho-tyrosine (pY) containing target molecules. Delineation of these binding sites as essentially short linear peptides (both structurally and functionally) has led to the suggestion that the activity of these signalling complexes may be manipulated by the development of relatively simple peptide reagents. This review examines the range of possibilities open on this approach and the extent to which positive results have already been realised. PMID- 8730510 TI - The stoichiometry of expression of protein components of the stimulatory adenylyl cyclase cascade and the regulation of information transfer. AB - Quantitative analysis of the proteins which compromise the stimulatory arm of the adenylyl cyclase cascade indicate that the adenylyl cyclase catalytic component is usually the least highly expressed. The effects on both potency of agonist ligands and maximal output resulting from targetted alterations in expression levels of each element of this cascade are discussed. PMID- 8730511 TI - Rapid regulation of PDE-2 and PDE-4 cyclic AMP phosphodiesterase activity following ligation of the T cell antigen receptor on thymocytes: analysis using the selective inhibitors erythro-9-(2-hydroxy-3-nonyl)-adenine (EHNA) and rolipram. AB - The PDE2, cyclic GMP-stimulated, and the PDE4, cyclic AMP-specific enzymes provide the major, detectable cyclic AMP phosphodiesterase activities in murine thymocytes. In the absence of the cyclic GMP, PDE4 activity predominated (approximately 80% total) but in the presence of low (10 microM) cyclic GMP concentrations, PDE2 activity constituted the major PDE activity in thymocytes (approximately 80% total). The PDE4 selective inhibitor rolipram dose-dependently inhibited thymocyte PDE4 activity (IC50 approximately 65 nM). PDE2 was dose dependently activated (EC50 approximately 1 microM) by cyclic GMP and inhibited by erythro-9-(2-hydroxy-3-nonyl)-adenine (EHNA) (IC50 approximately 4 microM). EHNA was shown to serve as a selective inhibitor of PDE-2 activity as assessed from studies using separated PDE1, PDE2, PDE3 and PDE4 species from hepatocytes as well as human PDE2 and PDE4 enzymes. EHNA completely ablated the ability of cyclic GMP to activate PDE2 activity, whilst having a much smaller inhibitory effect on the unstimulated PDE2 activity. EHNA exhibited normal Michaelian kinetics of inhibition for the cyclic GMP-stimulated PDE2 activity with Hill plots near unity. Apparent negative co-operative effect were seen in the absence of cyclic GMP with Hill coefficients of approximately 0.3 for inhibition of PDE2 activity. Within 5 min of challenge of thymocytes with the lectin phytohaemagglutinin (PHA) there was a transient decrease (approximately 83%) in PDE-4 activity and in PDE2 activity (approximately 40%). Both anti-TCR antibodies also caused an initial reduction in the PDE4 activity which was followed by a sustained and profound increase in activity. In contrast to that observed with PHA, anti-TCR/CD3 antisera had little effect on PDE2 activity. It is suggested that, dependent upon the intracellular concentrations of cyclic GMP, thymocyte cyclic AMP metabolism can be expected to switch from being under the predominant control of PDE4 activity to that determined predominantly by PDE2 activity. These activities may be rapidly and differentially regulated following ligation of different cell surface receptors. PMID- 8730512 TI - Synergistic activation of cAMP and calcium on cAMP-response-element-mediated gene expression in GH3 pituitary tumor cells. AB - Signals responsible for expression of the vasoactive intestinal peptide (VIP) stimulated prolactin gene in GH3 pituitary tumor cells were examined. Transfection with a deoxyribonucleic acid (DNA) construct containing the chloramphenicol acetyltransferase (CAT) gene fused to the 2.5-kb prolactin 5' upstream regulatory sequence indicated that VIP stimulated CAT expression. However, this effect could not be mimicked by 8-bromo-cyclic adenosine monophosphate (8-Br-cAMP), and was inhibited by the L-type Ca(2+)-channel blocker verapamil. While KCl had little effect on CAT activity, combined treatment with KCl and 8-Br-cAMP synergistically activated CAT expression. Potentiation between KCl and 8-Br-c-AMP was also seen with c-fos messenger ribonucleic acid (mRNA) expression. In addition, KCl and 8-Br-cAMP synergistically activated cAMP response element (CRE)-mediated CAT expression, and the synergism was abolished by verapamil. In the presence of okadaic acid, cAMP had no significant activation on CRE-driven CAT expression, whereas KCl-stimulated CAT expression was greatly potentiated. These results indicate that cAMP and Ca2+ synergistically activated CRE-driven gene expression through non-overlapping phosphorylation events in GH3 cells. PMID- 8730513 TI - Adrenoceptor heterogeneity in human white adipocytes differentiated in culture as assessed by cytosolic free calcium measurements. AB - Changes in intracellular calcium concentration [Ca2+]i in response to norepinephrine (NE) and to various adrenergic agonists were monitored by dual excitation microfluorimetry in single human adipocytes differentiated in culture and loaded with fura-2 acetoxymethyl ester (fura-2 AM). The addition of NE elicited increases in [Ca2+]i that were depending on the cell, (1) either rapid (time to peak: 9 +/- 3 s), large, and transient; or (2) slow (time to peak: 125 +/- 8 s), small, and sustained. The rapid and large [Ca+]i response, which was inhibited by 90% by the alpha 1-antagonist prazosin and only by 20% by the non specific beta antagonist (-)-propranolol, was considered to be mediated by the alpha 1-adrenoceptor. In fact, an alpha 1A-adrenoceptor was found to be expressed in human white adipose tissue. Consecutive additions of beta-agonists specific for each subtype of alpha-adrenoceptor enabled the characterization of four cell populations with different response patterns: 47% of the cells had alpha 1- and beta 1-, beta 2- and beta 3-induced [Ca2+]i responses; 29% had only beta 1-, beta 2-, beta 3-responses; 14% had alpha 1- and beta 3-responses, and 10% had only an alpha 1-response. Taken together, these results show that in differentiated human adipocytes: (1) alpha 1- and beta-adrenergic stimulations induce [Ca2+]i increases with different kinetics and amplitudes; (2) there is a beta 3 adrenergic response similar to the beta 1- or beta 2-adrenergic responses; and (3) there is a marked adrenoceptor heterogeneity. PMID- 8730514 TI - The role of protein kinase C in activation and termination of mitogen-activated protein kinase activity in angiotensin II-stimulated rat aortic smooth-muscle cells. AB - Mitogen-activated protein (MAP) kinases are a family of serine/threonine kinases activated by both tyrosine kinase and G-protein-linked receptor agonists. In rat aorta vascular smooth-muscle cells (VSMC), vasoconstrictors, angiotension II (AII), and alpha-thrombin (alpha-thr), as well as platelet-derived growth factor beta beta (PDGF) stimulated the tyrosine phosphorylation and activation of MAP kinase in a time- and concentration-dependent manner. Pre-treatment of cells with the protein kinase C (PKC) inhibitor Ro-318220, inhibited the initial increase in tyrosine phosphorylation of MAP kinase in response to vasoconstrictors, suggesting the involvement of PKC. Four isoforms of PKC were identified in VSMC by western blotting: alpha, beta, epsilon, and zeta. Downregulation of PKC alpha and PKC epsilon isoforms following chronic phorbol myristate 12, 13-acetate (PMA) pre-treatment resulted in the abolition of AII-stimulated MAP kinase activation. Selective downregulation of PKC alpha following pre-treatment with bryostatin 1 did not affect AII-stimulated MAP kinase. Preincubation of cells with Ro-318220 enhanced the activation of MAP kinase at later time points. In addition, Ro 318220 pre-treatment inhibited the induction by AII of a novel transcriptionally regulated phosphatase, MAP kinase phosphatase-1 (MKP-1). However, AII-mediated activation of MAP kinase was not prolonged by cycloheximide pre-treatment and was not maintained indefinitely by Ro-318220. These results demonstrate a specific role for the Ca(2+)-independent PKC isoform, PKC epsilon, in the activation of MAP kinase in response to vasoconstrictors, and suggest that PKC-mediated induction of MKP-1 plays no role in the termination of transiently activated MAP kinase. PMID- 8730515 TI - Uncoupling of the pathways which link MAP kinase to c-fos transcription and AP-1 in response to growth stimuli. AB - The v-Src oncoprotein induces mitogenesis and transformation of cells through multiple effects on diverse signalling pathways that are influenced by the cellular context in which v-Src is expressed. Here we have examined the effects of a temperature-sensitive (ts) v-Src on transcription of the c-fos proto oncogene, in serum-deprived and growing Rat-1 fibroblasts. We have also considered the role of mitogen-activated protein (MAP) kinase, a known mediator of ternary complex formation at the c-fos serum response element (SRE), which results in transcriptional enhancement in response to growth factors. In cells exponentially growing in the presence of serum, activation of v-Src stimulated MAP kinase and c-fos transcription. In cells made quiescent by serum deprivation, however, v-Src did not induce a c-fos transcriptional response, nor was there stimulation of ternary complex formation, despite normal activation of MAP kinase. Thus, activation of MAP kinase and stimulation of c-fos transcription and ternary complex formation are uncoupled in the absence of serum growth factors. Stimulation of c-fos by v-Src in growing cells, however, coincided with formation of a complex with an oligonucleotide spanning the c-Sis-inducible element (SIE) upstream from the SRE, suggesting that the signal transduction and activator of transcription (STAT) family of transcription factors, which bind here, may function in response to the v-Src oncoprotein. During these studies, we also observed that addition of fresh serum growth factors to growing Rat-1 fibroblasts expressing ts v-Src at the restrictive temperature resulted in substantially impaired activation of MAP kinase. This interference with normal growth factor signalling implies that catalytically inactive Src acts in a dominant negative manner by blocking normal activation of MAP kinase, although not at the expense of c-fos transcription. Thus, serum-induced c-fos transcription can also occur in an MAP kinase-independent manner. PMID- 8730516 TI - Chloride-channel block inhibits T lymphocyte activation and signalling. AB - Both large- and small-conductance chloride (Cl-) channels have been found in human T lymphocytes; however, apart from possible roles in mediating regulatory volume decrease, their functions are not understood. We have used patch-clamp electrophysiology, Ca2+ spectrofluorometry, and Western blot assay for phosphotyrosine to investigate the effects of blocking Cl- channels on proliferation and on specific events in the activation of normal human T cells. Four chemically distinct Cl- channel blockers inhibited both the small conductance Cl- channels and phytohemagglutinin (PHA)-induced lymphocyte proliferation in a similar dose-dependent manner; their order of potency was 5 nitro-2(3-phenylpropylamino)-benzoic acid (NPPB) > 4,4'-diisothiocyano-2,2' disulfonic acid (DIDS) > flufenamic acid >> IAA-94. The Cl- channel blockers inhibited both the PHA-induced mobilization of Ca2+ and the rapid tyrosine phosphorylation of several polypeptides. Cell proliferation was not rescued by the Ca+ ionophore ionomycin or by addition of exogenous interleukin-2 (IL-2). Moreover, the blockers also inhibited phosphotyrosine expression in IL-2-treated, activated lymphoblasts. Thus, our results support a role for Cl- channels in early, PHA-evoked signalling and in later, II-2-dependent stages of lymphocyte activation and proliferation. PMID- 8730517 TI - Reactivity to smoking cues: mediating roles of nicotine dependence and duration of deprivation. AB - This study examined the roles of nicotine dependence and duration of smoking deprivation as mediators of smoking cue reactivity in 117 individuals enrolled in treatment. Participants were randomly assigned to 15, 90, or 180 min of deprivation; all were exposed to smoking cues (observing a research assistant smoke a cigarette) while a variety of self-report, behavioral, and psychophysiological measurements were obtained. Results indicated that in general, psychophysiological reactivity to smoking-relevant cues was greater with increasing deprivation; heart rate analysis revealed an interaction between nicotine dependence and duration of deprivation. Smoking desire, negative affect, physical withdrawal symptoms, and other ratings were interrelated during cue exposure, and generally demonstrated predicted relationships with the independent variables. Multiple regression analysis revealed that desire to smoke during cue exposure was predicted by a combination of psychophysiological, self-report, and demographic variables. These findings have implications for methodological considerations and theoretical underpinnings of smoking cue reactivity research. PMID- 8730518 TI - Female PCP-using jail detainees: proneness to violence and gender differences. AB - Previous studies indicate that PCP users have different characteristics from other drug users and that female PCP use is more common than use among males. Furthermore, there is evidence that those who respond to PCP with violence may differ from those who do not. This study attempted to examine comprehensively the psychological, behavioral, and background factors among female jail inmates that may contribute to a PCP preference and subjects' perception of various behavioral states while using PCP. Female PCP users were further examined relative to male PCP users to differentiate them on the basis of these perceptual factors. A distinction was further made between females and males prone to PCP-induced violence and those who do not become violent with respect to the above psychological and behavioral measures. Our results showed differences between male and female PCP users that are discrepant with the assumption that men and women perceive similar drug-related experiences. In particular, female PCP using subjects reported more dysphoria and aggressiveness when not using PCP, while male subjects were more likely to report aggressive behavior and dysphoria under the influence. Overall, these results suggest that males who prefer PCP may be self-stimulating and females who prefer PCP may be attempting to self-medicate. PMID- 8730519 TI - Long-term associations of helpful and harmful spousal behaviors with smoking cessation. AB - It has repeatedly been demonstrated that increased levels of social support are positively associated with a smoker's likelihood of achieving abstinence. It remains unclear, however, what specific support behaviors influence smoking cessation. This study explores the effects of supportive and undermining behaviors experienced by subjects (N = 624) in the context of a romantic/marital relationship on a number of smoking cessation outcomes. The associations of social support behaviors with one-, two-, and three-year abstinence and recycling and making quit attempts at 6, 12, 24, and 34 months were examined. Frequencies of both supportive and undermining behaviors predicted the dependent variables under study. Subjects whose spouses demonstrated fewer undermining behaviors were more likely to be successful continuous abstainers, whereas participants' recycling and making quit attempts seemed to be best predicted by a greater frequency of supportive behaviors. Initial success at cessation might be associated with increased supportive behaviors by a spouse, whereas lower rates of undermining behaviors might be associated with long-term maintenance. Long term tailored feedback to spouses of smokers wanting to quit might be helpful. PMID- 8730520 TI - Social influences on young adults' alcohol consumption: norms, modeling, pressure, socializing, and conformity. AB - This study aims to assess which types of social influence are correlated with young people's (15-24 years) heavy drinking (six or more glasses) in public drinking places during the weekend. Drinking in public drinking places can be defined as a "timeout" situation. Therefore we assumed that situational factors (e.g., importance of socializing and direct pressures on drinking) would contribute more to the explained variance than variables indicating cognitive social influences (e.g., social norms and modeling). Stepwise regression analyses showed that in total 25% of the variance was explained by social norms of family and peers (15%), importance of socializing in drinking situations (7%), modeling (2%) and group size (1%). The results show that both a cognitive factor and a situational factor appear to be most strongly correlated with young people's frequency of heavy drinking in public drinking places. Within the category of situational influences those variables indicating direct social pressures were only weakly related or not significant. Studies focusing on measuring the impact of social influences may profit from including the concept of the importance of socializing and conformity as an additional factor. PMID- 8730521 TI - Measuring drug urges by questionnaire: do not balance scales. AB - Measures designed to assess self-reported drug urges are routinely balanced; that is, they contain a relatively equal number of positively and negatively worded items. Using smoking urges as an example, we explored the effect of item wording on responses to self-report items. One hundred sixteen cigarette smokers responded to 32 positively worded statements and their 32 negatively worded counterparts adapted from the Questionnaire of Smoking Urges. For 59% of the paired statements, the negatively worded item was scored higher than the positively worded item. Significant mean differences (item means for 56% of the paired statements were significantly different at the .01 level) and only moderate correlations (mean = .54; range = -.04 to .79) between item pairs were found. Balanced scales may have greater costs than benefits in the assessment of drug urges. PMID- 8730522 TI - Measuring cocaine effect expectancies among therapeutic community inpatients. AB - Evidence has accumulated that beliefs about alcohol contribute to the etiology and maintenance of different patterns of drinking and drinking-related behavior. This research focus has been expanded from alcohol to include marijuana and cocaine. The present study was designed to improve upon earlier efforts to measure the domain of cocaine effect expectancies, which were based on samples of college students. Confirmatory factor analyses of the 24-item Cocaine Effect Expectancy Questionnaire (CEEQ) were conducted using a sample of 231 therapeutic community inpatients in the northeastern United States. The suggested factor structure was not confirmed. Empirical support was found for a 4-item Positive Effects scale. This scale did not have a significant effect on treatment participation. PMID- 8730523 TI - Relationship of depression with measures of social functioning in adult drug abusers. AB - The present study describes the relationship between depression and several measures of social functioning--including employment, criminal activity, incarceration, marital functioning, and alcohol and drug use--in a population of adult drug abusers. Our investigation extends past work in this area by specifically investigating the effects of depression (as opposed to simple substance use) on social and interpersonal functioning. Predictably, elevated levels of depression were associated with increased use of hard drugs and alcohol, greater levels of institutionalization, reduced attendance at work or school, and lower overall rates of marital satisfaction. Consistent with previous reports, level of marijuana use was not related to severity of depression. It appears that depressed substance abusers experience significantly more social, vocational, and interpersonal dysfunction than their nondepressed counterparts. It is proposed that the efficacy of existing treatment programs for adult drug abusers will be enhanced through the addition of strategies to assess and ameliorate depression. PMID- 8730524 TI - Bias in reported body weight as a function of education, occupation, health and weight concern. AB - Comparison of self-reported and measured weights in a working population of 2046 men and 2393 women revealed systematic underreporting of 1.3% in men and 1.7% in women. Underreporting was significantly related to weight, height, and current participation in a weight reduction program in both men and women. In men only, it was also related to age, education, history of weight-control attempts, and history of weight-related health conditions. Overall, however, these predictor variables accounted for a small fraction of the variance in underreporting and the correlation between measured and self-reported weight was very high (rs = .99). It is concluded that self-reported body weight is an excellent approximation of actual weight across a broad range of population subgroups. PMID- 8730525 TI - Cognitive therapy techniques in continuing care planning with substance-dependent patients. AB - Cognitive therapy techniques were used with substance-dependent inpatients in order to increase patient involvement with continuing care/aftercare resources. Patients receiving cognitive techniques were compared to an earlier grouping of patients with whom cognitive therapy was not used. Results indicated that cognitive therapy significantly improved both referral acceptance and initial patient contact with continuing care agencies. The complexity of continuing care compliance and opportunities for further research are discussed. PMID- 8730526 TI - Smoking topography in tobacco chippers and dependent smokers. AB - Although most cigarette smokers exhibit signs of tobacco dependence, a subset of this population, referred to as tobacco chippers, does not show characteristic signs of dependence. Few studies have attempted to characterize differences between these groups of smokers. The purpose of the present study was to examine smoking topography in chippers (CH) and dependent smokers (DS). Topographical variables including puff number and duration, and intercigarette interval were examined in seven CH and seven DS under both laboratory and naturalistic conditions. Saliva nicotine, cotinine, and thiocyanate, as well as expired air carbon monoxide (CO) levels were also measured. The results indicate that there were no differences in smoking topography between CH and DS, except those that would be expected based on selection criteria. Although there were differences between groups on pre- and postsmoking CO and saliva cotinine levels, there were no differences in change scores from pre- to postsmoking on these measures. Additional studies will need to be done in order to completely characterize differences between CH and DS. PMID- 8730527 TI - Brief assessments of dietary behavior in field settings. AB - Our research team is involved in ongoing research in both worksites and medical office settings. These settings offer great potential for reaching individuals who would not otherwise participate in health promotion, but they also place considerable constraints on assessment time and efforts, especially if one's goal is to attract a high and representative proportion of employees or patients. This paper reports on our experience with measures of dietary behavior in these two settings. We found it problematic to collect detailed assessments such as 4-day food records or comprehensive food frequency/history checklists in worksites or medical office settings using population-based samples. Instead, we recommend and provide data on the utility of a dietary-fat screening instrument, and on the Food Habits Questionnaire (FHQ-Kristal, Shattuck, & Henry, 1990), a brief measure of dietary behaviors associated with high-fat eating patterns. The FHQ, in particular, was found to correlate well with other more costly and time-consuming methods of assessment, to be reliable and responsive to intervention effects, and to provide behavioral targets for intervention. The strengths and limitations of these measures for tailoring intervention and assessing outcomes are discussed. PMID- 8730529 TI - The organizational implications of smokeless tobacco use in the lumber mill industry. AB - Although much is known about the characteristics of employees who smoke cigarettes, very little is known about workers who use smokeless tobacco. The current study was designed to understand the characteristics of smokeless tobacco users in relation to their performance at work and compare them with smokers and former tobacco users. Data were collected via interviews and questionnaires from a random sample of employees working at Pacific Lumber Company (N = 146), the largest single-site lumber mill in California. A total of 63 smokeless tobacco users (21 of whom also smoked cigarettes), 43 cigarette smokers, and 40 employees who had successfully quit using tobacco (34 of whom previously used cigarettes only) provided information about their health behavior, quality of work life, and performance at work. Analyses revealed that smokeless tobacco users reported less healthful sleep patterns, drank alcohol more often, were intoxicated more often, reported less job satisfaction and organizational commitment, and reported that both chewers and smokers do not work as hard and take more breaks than do tobacco free employees (quitters). Specific differences among chewers-only, smokers-only, smokers-and-chewers, and quitters are presented. Results suggest the organizational value of developing worksite cessation programs for smokeless tobacco users. PMID- 8730528 TI - Condom use among gay/bisexual male substance abusers using the timeline follow back method. AB - Sexual risk for HIV transmission under the influence of alcohol and/or other drugs is not simply a cause-effect relationship: not everyone who drinks or uses other drugs has unprotected sex. The purpose of this study is to explore differences between substance using gay/bisexual men who use condoms during anal sex from those who do not. These differences are identified by comparing men whose anal sex while under the influence of alcohol and/or drugs is consistently protected to men whose anal sex while under the influence of alcohol and/or drugs is consistently unprotected. Gay/bisexual men entering substance abuse treatment at a gay-identified agency in San Francisco were recruited to complete surveys and to be interviewed about sexual behavior, substance use, and related variables using an extended version of the Timeline Follow-back (TL). The TL procedure uses a blank calendar form and a series of questions to cue recall of drinking, drug use, and anal intercourse on each of the 30 days prior to the last date of alcohol and/or drug use. Men whose anal sex while under the influence of alcohol and/or drugs is consistently unprotected were significantly more likely to report having less than a college education (p = .04), more likely to have an income of less than $20,000 (p = .01), more likely to use amyl nitrite (p = .01) and cocaine (p = .02), and more likely to report a higher frequency of anal sex (p = .007). In addition, they were less likely to approve of sex without love (p = .003), less likely to perceive that safer sex is the community norm (p < .001), and less likely to have encouragement from friends to practice safer sex (p = .001). However, HIV status did not differentiate between the two groups. These two groups provide clear and interesting contrasts in terms of behavior, thus comparisons of the factors influencing sexual safety in these subgroups may enhance our understanding of risk taking. A better understanding of possible mediating variables can be important both in guiding future research in this area and in formulating intervention strategies to target gay men who drink or use drugs in combination with sexual activity. PMID- 8730530 TI - Substance use and impulsive behaviors among adolescents with eating disorders. AB - Results of past research suggest that the existence of bulimic behaviors (binge eating and/or purging) may be an indicator of increased likelihood of substance use. We investigated incidence of substance use among adolescent girls (mean age = 15.4 years) with anorexia nervosa (n = 59) or bulimia nervosa (n = 58). The incidence of substance use among girls with anorexia nervosa was low, particularly after removing those anorexic adolescents with bulimic symptoms. Nearly one-third of girls with bulimia nervosa had smoked tobacco cigarettes, had used marijuana, and were drinking alcohol at least weekly. Among those exhibiting bulimic symptoms, increased experience with use of different substances was related to increased incidence of attempted suicide, stealing, and sexual intercourse but was unrelated to age or incidence of intentional self-harm behavior. Our findings are discussed in relation to the results of past research and the clinical implications of our data. PMID- 8730531 TI - Assessing the needs of the severely mentally ill: cultural and professional differences. AB - International interest is currently focused on the effectiveness of mental health services in helping the severely mentally ill. An important outcome measure is change in the level of individual need. Little research has been done into how mental health professionals prioritise needs. This question was addressed by undertaking a survey of mental health professionals in cities in India, Australia and England. Similar priorities were found in all survey sites, and across hospital and community staff, though differences were found due to professional background. This suggests that there is some cross-cultural consistency in prioritising needs, but the choice of profession to assess need may influence the way in which needs are assessed. PMID- 8730532 TI - Insight and general public attitude on psychotic experiences in Hong Kong. AB - Psychosis has been viewed with vastly different attitudes in different contexts. Medical professionals frequently assume that patients should have adequate insight into their abnormal experience. "Lack" of insight has been regarded as a characteristic feature of schizophrenic psychosis. However, these experiences have frequently been construed from a non-medically related perspective by the general public. We employed a case-vignette approach to study the opinion of a general public sample concerning attribution, intervention needs and concordance with medical professionals towards psychotic experience. Ninety eight healthy Chinese volunteers were recruited. Results showed that public opinion only agreed partly with the conventional medical model. Although most subjects agreed that the experience described in the case-vignette was psychological, they did not relate this to psychiatric treatment. Subjects having past contact with psychiatric patients showed a higher degree of agreement with the medical perspective than subjects without past contact. Potential implications of our results on management and health education are discussed. PMID- 8730533 TI - The mental health pattern of migrants: is there a euphoric period followed by a mental health crisis? AB - Sluzki's 1986 mental health model of the migratory process was tested with migrants (both refugees and immigrants), to New Zealand. Its central feature, suggesting an initial symptom free and euphoric phase after arrival in the country of settlement, followed by a crisis stage, was examined for 129 Southeast Asian refugees, 57 Pacific Island immigrants and 63 British immigrants. A questionnaire and the Hopkins Symptom Checklist-25, in English and in three Southeast Asian language translations, were administered face-to-face. All respondents had arrived in New Zealand within the last 15 years. The findings did not support Sluzki's model. Refugees and immigrants in the group with less than six months of residence were not symptom free. Neither did the group with six months to six years residence demonstrate a deterioration in mental health. However, mean depression levels were slightly lower for those who had lived in New Zealand for over six years, suggesting that mental health may improve the longer both refugees and immigrants reside in the host country. PMID- 8730534 TI - Does acculturation influence psychosocial adaptation and well-being in Native Hawaiians? AB - Native Hawaiians have suffered higher disease rates and achieved lower educational and economic levels than other ethnic groups living in Hawaii. This study tests the hypothesis that cultural affiliation protects against psychosocial pathology in Hawaiians. Subjects included 172 Hawaiians of varying blood quantum and 92 non-Hawaiians. The Na Mea Hawai'i, a rationally derived, empirically validated measure of Hawaiian acculturation was administered to all subjects in addition to psychological tests which included 19 visual analogue scales that measure both state and trait variables. The Na Mea Hawai'i proved excellent at differentiating Hawaiians from non-Hawaiians (p = .0001), and it correlated well with blood quantum (r = 0.31, p = .0001). Low acculturation scores among Hawaiians did not correlate with any measure of discontent or psychosocial pathology irrespective of blood quantum. We conclude that Na Mea Hawai'i is a valid measure of Hawaiian acculturation, and that adaptation of the Hawaiians in our sample has been unaffected by level of acculturation. PMID- 8730535 TI - Religion, ethnicity and obsessive-compulsive disorder. AB - Obsessive-compulsive disorder (OCD) is seen in many contrasting cultures but it is not known if the form of the disorder varies between these cultures. There have been anecdotal case reports where religion appeared to play a significant aetiological role in the disorder but the relationship between religion and OCD has not previously been systematically studied. This study was a retrospective, casenote study comparing the country of birth and religious affiliation of three groups of 50 patients. Its aim was to investigate the aetiological role played by religion in the development of OCD. The groups were patients with OCD from a specialist behavioural-cognitive unit, patients assessed in a specialist psycho dynamic psychotherapy department and patients attending a general adult psychiatry outpatient department. More patients with OCD affiliated themselves with a religion as opposed to either of the other two groups. This difference disappeared when the type of religion was taken into account so that no conclusive relationship between OCD and religion could be identified. The findings do not diminish the importance of religion in the development of OCD in some individuals and suggest that future research in this area should include examination of the rigidity of upbringing and personal perception of the experience of strict rules or imposed religious practices. PMID- 8730536 TI - DSM-III mental disorders in general medical sector: predictors of outcome. AB - In 1987, a multicentric research was carried out in Emilia-Romagna, an administrative area in northern Italy. Nine hundred and forty one people in the general medical sector were contacted to enter a two-stage procedure to identify DSM-III cases. Cases were recontacted for 3-month, 1-year and 2-year follow-ups. Eighty cases completing at least the one-year follow-up were analysed. The sample was split into "still-cases" and "no longer-cases" depending on the persistence of a DSM-III diagnosis at the last follow-up. Both clinical and social features proved to be predictors of the outcome. A disorder lasting one year or longer, past recurrences, Axis I comorbidity, older age, being married and having children were the features which at the first assessment predicted an unfavourable outcome. Having children and past episodes proved to be the best predictors of remaining a case in a stratified logistic regression model. The persistence of major difficulties during the follow-up period was also correlated to a "still-case" status. PMID- 8730537 TI - Neuroticism in a family practice population in India. AB - This study was conducted to assess the prevalence of neuroticism and its correlation with the various sociodemographic factors and presenting complaints among patients attending a family practice outpatient clinic. One hundred and forty two consecutive patients attending the outpatient clinic were administered PGI Health Questionnaire N2. Higher incidence of neuroticism was observed in middle age group population, in females (especially housewives), unskilled workers, patients with lower educational status and those belonging to small and nuclear families. A significant number of such patients had presented with vague symptoms, were visiting the clinics frequently and had associated chronic diseases. The results suggest that patients visiting the family practice clinics frequently with multiple somatic complaints with no suggested organic basis should undergo an in-depth psychiatric evaluation. PMID- 8730538 TI - Brief report: families of rural mentally ill and treatment adherence in district mental health programme. PMID- 8730539 TI - The International Classification of Impairments, Disabilities and Handicaps (ICIDH)--results and problems. AB - In 1980, the World Health Organization (WHO) published the International Classification of Impairments, Disabilities and Handicaps (ICIDH). Although these three concepts of impairment, of disability, and especially of handicap have been criticized for many reasons, the ICIDH has given rise to extensive research and other activities world-wide, resulting in more than a thousand publications. The ICIDH has been translated into numerous languages. The classification itself, developed for trial purposes, shows the large variety for the items of each basic term. Practical experience, however, has shown that the ICIDH cannot be applied without problems. This is especially true for the handicap concept. PMID- 8730540 TI - The economics of independent living: efficiency, equity and ethics. AB - People with disabilities are routinely denied the exercise of choice in their daily lives. There are strong efficiency arguments for the promotion of greater choice and autonomy for disabled people. There are equally strong moral arguments for an investment in the capabilities of disabled people to allow them to participate in both the educational system and the labour market. This investment will not come cheaply nor will the pay-off always be of such magnitude to justify the expenditure on narrow cost-benefit criteria. Those who value efficiency above everything else must, however, set out the system of justice implied by such a choice. The conclusions of a narrow efficiency argument may turn out to be unacceptable to the majority of citizens. Likewise, however, those who value equity at all costs must consider the implications of their approach for individual freedom, economic growth and technical efficiency. This paper is an attempt to explore the meaning of efficiency and equity in the context of independent-living programmes for people with disabilities. PMID- 8730541 TI - Transition from school to work of students with developmental disabilities and mental retardation: an Israeli perspective. AB - The present research was conducted in four special education schools for students with developmental disabilities and mental retardation in the Haifa area. The total number of subjects was 72. There were 45 (63%) males and 27 (37%) females. The mean age was 18.1 years (range: 16-19.5). The mean IQ (WISC) was 66.3 points (range: 40-85). The vocational placement at the end of 6 months after leaving school was recorded for each school graduate. A total of 53 teachers--school principles, counsellors, specialized teachers and speech and occupational therapists--filled out specially designed questionnaires covering three areas: (1) students' work programme experience; (2) teachers' ratings of final year students on: academic studies, social behaviour, independence and vocational skills as well as teachers' recommendations, given towards the end of the final year at the school, for the immediate plan of action for each of the graduating students; (3) teachers' ratings of parental involvement with the educational programme of each student. Background variables included IQ, age and gender. The major finding was that 6 months after graduation, one-third of special school graduates were not working, were not enrolled in any formal programme and were idle at home. The variables that were found to correlate with successful placement were students' prior work experience, teachers' ratings of students achievements and their recommendations for each student, and parental involvement. PMID- 8730542 TI - Outpatients in neurological rehabilitation. AB - This paper discusses the experience of working within a multidisciplinary neurological clinic in a regional rehabilitation centre. The number and types of interventions are analysed which confirm the importance of having a wide range of rehabilitation professionals available in or attached to such clinics. The implications of these findings are discussed with regard to the range of professionals required and the study has produced evidence that would support the establishment of certain specific disability clinics, particularly spasticity, orthotic and memory clinics. The need for specific disease-orientated clinics, particularly for multiple sclerosis and head injuries, is also discussed. A questionnaire was sent to all new attenders at the Newcastle clinic seeking patient's views on the clinic format. There was general support for a multidisciplinary approach to outpatients and the majority of people felt that their needs had been properly addressed. However, a significant minority felt intimidated by a large number of professionals seen simultaneously and felt that their full range of problems had not been discussed. The patient's preference was for a drop-in clinic when they could attend without appointment to discuss a particular problems with the relevant professional. The least popular format was to be seen simultaneously by a range of different rehabilitation professionals. PMID- 8730543 TI - Right cerebral hemisphere damage: incidence of language problems. AB - While dysphasia is rare after right cerebral hemisphere damage (RHD), there is now a body of evidence to suggest that language and communication problems can occur. This paper reports the findings of a pilot study that estimated the incidence of non-dysphasic language and communication problems following RHD. Eleven subjects completed language assessments within 2 weeks of onset and again 3 months later. The results show that language impairment can follow a unilateral RHD for 50% of the sample, and that there are marked difficulties for 20%. Furthermore these problems do not resolve spontaneously over a 3 month period. The case for speech and language therapy intervention for this patient group is discussed. PMID- 8730544 TI - Incidence of secondary complications in spinal cord injury. AB - In order to document the incidence of secondary complications following spinal cord injury (SCI), data were collected from 348 patients with post-acute SCI. More than 95% of these patients reported at least one secondary problem, and more than half (58%) reported three or more. Statistical analyses indicated that, in general, the number and severity of complications varied with the time since injury. Obesity, pain, spasticity, urinary tract infections, and pressure sores were common. Issues of social integration were also identified. The results suggest that improvement is needed in practices related to prevention and follow up. PMID- 8730545 TI - Reliability of the Modified Ashworth Scale in the assessment of plantarflexor muscle spasticity in patients with traumatic brain injury. AB - Although the Modified Ashworth Scale (MAS) is commonly used to assess the severity of muscle spasticity for ankle plantarflexors, its reliability has only been established for elbow muscles. Interrater reliability, intrarater reliability and temporal (between-days) reliability were examined in this study. Also, interrater reliability for use of the scale with plantarflexors was compared with reported results from the measurement of elbow flexors. Thirty adult volunteers with traumatic brain injuries participated. There were 20 men and 10 women; the mean age was 28.3 years (SD = 10.8). Two physical therapists used the MAS to score the subjects independently. Measurements were repeated to yield multiple scores for intrarater reliability assessment. Twenty-one of the subjects returned individually on separate days to be measured again, so that temporal reliability could be assessed. Spearman's correlation coefficients were 0.73 for interrater reliability 0.74 and 0.55 for intrarater reliability, and 0.82 for temporal reliability. Overall, reliability of the MAS for assessing plantarflexor spasticity in patients with traumatic brain injury was found to be minimally adequate to support its continued use. However, interrater reliability was less than that which has been reported for elbow flexors, and intrarater reliability findings were mixed. PMID- 8730546 TI - Definition and construction of a single switch control environment for music composition. PMID- 8730547 TI - Choice and daily activities for people with disabilities. PMID- 8730548 TI - Molecular and cytogenetic abnormalities in acute myeloid leukaemia and myelodysplastic syndromes. AB - With the use of molecular techniques it is now possible to define even subtle chromosomal abnormalities and the fusion products resulting from translocations. Defined clinical correlations can now be made and prognostic implications are already found. For instance, patients with AML carrying t(8;21), t(15;17) or inv(16) have a better prognosis for long-term survival. This is also illustrated by Figure 1, which shows data of the Dutch HOVON AML study. The definition of patients with a bad or good prognosis has already resulted in the adjustment of treatment protocols. In the near future, with the use of more defined molecular techniques, we might be able to characterize the chromosomal abnormality of each patient, to individualize his treatment and to recognize very early relapses. PMID- 8730549 TI - Special cytological subtypes of acute myeloid leukaemias and myelodysplastic syndromes. AB - The recognition of distinct cytoimmunological subsets of pre-leukaemia and overt AML has been accomplished by morphological, immunological and ultrastructural studies. In many cases, a strong association has been documented between distinctive cytological features and specific chromosome changes. The primary genetic event underlying malignant transformation was also elucidated in a number of acute leukaemias and, as a matter of fact, assessment of these biological parameters has now an established role in the diagnostic work-up and in the monitoring of residual disease. On a more general basis, biological research in MDS is gradually clarifying the fundamental pathophysiological mechanisms of altered cell growth, and differentiation and therapeutic decision making in leukaemia is becoming increasingly dependent on the precise characterization of blast cells. Further refinement of the cytoimmunological classification of acute leukaemias and MDS is warranted in order to provide the physician with an updated framework of reference for the categorization of these heterogeneous haematological disorders and to improve the reproducibility of current morphological diagnosis among different centres (Castoldi et al, 1993). PMID- 8730550 TI - Immunophenotyping of AML and MDS and detection of residual disease. AB - Immunophenotyping improves both accuracy and reproducibility of the FAB classification and is considered particularly useful for identifying poorly differentiated FAB subtypes of AML, such as AML with minimal differentiation (M0), microgranular promyelocytic leukaemia (M3V), and megakaryoblastic leukaemia (M7). Immunological studies of myeloid leukaemic blasts has become critical also in identifying biphenotypic leukaemias and AML expressing lymphoid-associated markers (Ly+ AML). At present, while the prognostic value of individual antigen expressions is still controversial, due to technical questions, the immunological detection of MRD seems to be important in monitoring AML patients in remission and, perhaps, in detecting leukaemic cell contamination into bone marrow or peripheral blood progenitor cells collected for autologous transplantation. In addition, the relationship established between genetic abnormalities and certain phenotypes within different FAB subtypes suggests that, in the future, immunophenotypical studies could be used for the screening of AML cases carrying specific genetic aberrations. Compared to acute leukaemias, little information is available concerning immunological patterns in MDS, and the role of the immunophenotype in diagnosis, subclassification, and prognosis of MDS is currently not well established. PMID- 8730551 TI - Risk of acute myelogenous leukaemia and myelodysplasia following cancer treatment. AB - Now that a substantial group of cancer patients has such a favourable prognosis, it has become increasingly important to evaluate the long-term complications of treatment. Of all late effects of treatment, secondary leukaemia is one of the most serious. Increased risk of AML has been observed both after RT and after CT; however, several types of CT have much stronger leukaemogenic properties than RT. Limited field radiation in the therapeutic dose range is associated with very little or no increased risk of leukaemia, which has been attributed to cell killing at the higher radiation doses. With respect to CT, two different syndromes of treatment-related AML have been recognized. Risk of alkylating agent related AML is highest in the 5-10 year follow-up period and seems to decrease afterwards. This type of leukaemia is often preceded by MDS, and is characterized by deletions of chromosomes 5 and 7. Leukaemias related to treatment with the topoisomerase II inhibitors are characterized by a short induction period, presentation as myelomonocytic or monocytic leukaemia (rather than MDS) and balanced chromosomal translocations involving bands 11q23 and 21q22. This review addresses the risk of secondary AML and MDS following treatment of HD, NHL, testicular cancer, ovarian cancer, breast cancer and paediatric malignancies. In patients with HD, the risk of AML is higher with an increasing number of mechlorethamine-procarbazine-containing cycles, a greater number of CT episodes, and after splenectomy. The majority of data shows that RT does not add to the leukaemia risk from CT, but this issue is still surrounded by some controversy. ABV(D)-treated patients have a very low risk of AML. Generally, patients with NHL, testicular cancer and breast cancer experience much lower risk of AML than patients with HD. NHL and breast cancer treatment regimens with cumulative cyclophosphamide doses of 20 g or less do not confer an appreciable increase of AML. Recently, strongly increased AML risk has been observed following autologous bone marrow transplantation and other dose intensification strategies. Risk factors for this excess remain to be defined. PVB treatment for testicular cancer is not followed by increased leukaemia risk, but modern etoposide-containing regimens do confer excess risk, of which the magnitude at conventional drug doses is not yet well known. High risk of leukaemia has been reported in children treated with epipodophyllotoxins. The leukaemogenic hazards of cancer treatment should be weighed against their therapeutic benefits. PMID- 8730552 TI - Treatment and prognostic factors in acute myeloid leukaemia. AB - Current induction chemotherapy regimens afford a complete remission in 60 to 80% of patients less than 60 years of age who are diagnosed with acute myelogenous leukaemia. Patients who undergo consolidation with high dose Ara-C or with autologous or allogeneic bone marrow transplantation can expect a long-term disease free survival of 30 to 45%. Which mode of consolidation is preferable is still being investigated in randomized studies. Many disease characteristics have been put forward as prognostic variables in treatment outcome and as understanding of the biology of AML has continued to expand, cytogenetic and molecular markers have assumed greater importance in disease characterization, but the ultimate role that these play in prognosis remains incompletely defined. Once AML has relapsed, bone marrow transplantation may result in prolonged disease free survival as compared to use of salvage chemotherapy regimens. Because the long-term survival of all patients diagnosed with AML is only in the order of 10 to 20%, new treatment approaches to this disease are required. Current clinical and research efforts have focused on modulation of the immune system, modulation of drug resistance phenotypes, and understanding of the response of leukaemic cells to haematopoietic growth factors. PMID- 8730553 TI - Treatment of acute promyelocytic leukaemia. AB - Currently available clinical results show that the combination of ATRA and anthracycline-Ara-C chemotherapy can slightly increase the CR rate in newly diagnosed APL, from about 80% (with chemotherapy alone) to more than 90%, and patients presenting with high leukocyte counts seem to benefit particularly from this combined therapy. ATRA followed by chemotherapy also reduces the incidence of relapse (particularly of early relapse) as compared to chemotherapy alone. However, treatment with ATRA is still complicated by the risk of hyperleukocytosis and potentially fatal ATRA syndrome, whose best preventive approach (addition of chemotherapy and/or dexamethasone) is still debated. Because some patients still relapse after ATRA plus chemotherapy, prognostic factors for relapse need to be precisely determined. They certainly include persistence or re-appearance of PML-RAR transcript during follow-up. Allogeneic BMT should be considered in those patients. Most of the patients who are not cured by the combination of ATRA and chemotherapy or by subsequent allogeneic BMT still die from their disease. This is due to the acquisition by APL cells of progressive resistance to ATRA, that appears to depend on induction of increased ATRA catabolism in APL cells. Current efforts aimed at overcoming this resistance (including intermittent ATRA schedules, synthesis of new retinoid compounds, utilization of inhibitors of cytochrome P450) are being developed. PMID- 8730554 TI - Treatment of childhood acute myeloid leukaemia. PMID- 8730555 TI - Treatment of the elderly patient with acute myeloid leukaemia. AB - Individuals of 60 years living in western countries generally have a mean life expectancy of 20 years at least. Therefore, when aged individuals present with AML, it is a necessity and a challenge to treat them as efficiently as possible. AML is mainly a disease of the elderly and accounts for more than 50% of its incidence among the general population. The treatment of older individuals with AML has remained difficult and its success is still limited. While in adults with AML of less than 60 years complete responses above 65% and survival rates of 35% are commonly obtained, progress in the treatment of elderly patients has been relatively small. As of today, approximately 50% of older patients may be induced into remission with chemotherapy, and, among these complete responders, only approximately 1 in 10 will survive free of leukaemia beyond 4 years after diagnosis. In fact, on one hand, these results represent the rationale and motivation for offering chemotherapy to the older population. On the other hand, they emphasize that major obstacles to better cure rates still exist. These stumbling blocks apparently relate to the restricted tolerance of older subjects to the exposure of chemotherapy and probably also a greater probability of unresponsiveness of the leukaemia to cytotoxic therapy. The haematopoietic growth factors still hold some promise and may improve outcome, but for the time being there is insufficient direct evidence to indicate a defined and established role. It is evident that new avenues should be pursued and trials specifically designed for elderly people with AML be conducted. These trials would need to address questions related to the choice of chemotherapeutic drugs (e.g. idarubicin versus mitoxantrone), their dose and schedule selection, the use of multidrug resistance modulators (to overcome intrinsic drug non-responsiveness), and the optimal clinical use of haematopoietic growth factors, including thrombopoietin. Since trials addressing specific questions regarding the development of treatment of elderly patients with AML have remained scarce, the initiation of these studies is sorely needed. One may hope that these clinical trials will provide some of the necessary answers and new clues, and will be useful to advance future therapy of elderly AML patients. PMID- 8730556 TI - Treatment and prognostic factors in myelodysplastic syndromes. AB - While MDS remains an enigmatic disease, substantial progress has been made in the elucidation of its origin and the better understanding of its natural course. The advent of newer molecular and cytogenetic techniques has tremendously improved the 'older' morphological and histopathological prognostic criteria. More refined scoring systems may ultimately allow for individualized treatment programmes which will better preserve quality of life, while at the same time offer improved chances for survival and cure. Much can be expected from newer cytokines, such as thrombopoietin, stem cell factor, interleukin-11 or of the combination of different cytokines and growth factors, to alleviate MDS-symptoms and to possibly alter the course of the disease. After the initial disappointment with differentiation inducers, the availability of newer agents and/of combinations may offer better perspectives for the future. Much interest will also be generated on the use of mdr-reversal agents in the attempts to improve on chemotherapeutic efficacy. Finally, while allogeneic transplantation still remains the only option for definite cure of the disease, the spectacular advances made in the use and manipulation of autologous peripheral blood haemopoietic stem cells probably constitute the best hope for brightening the grim outlook most MDS patients still have. PMID- 8730557 TI - Multidrug resistance in acute myeloid leukaemia. AB - Multidrug resistance represents a form of pleiotropic drug resistance that has a prognostic value in untreated AML. It may affect the outcome of current chemotherapy protocols. Therefore, MDR1 expression should be systematically investigated in prospective studies. In addition, restoration of drug sensitivity may be attempted by adding drug resistance modifying agents such as cyclosporins to standard chemotherapy. The clinical value of such an approach has to be established in currently ongoing phase III studies. PMID- 8730558 TI - Neuropsychology in psychiatric research: the missing link. PMID- 8730559 TI - Genitourinary medicine. PMID- 8730560 TI - Modern diagnosis and management of genital Chlamydia trachomatis infection. AB - Chlamydia trachomatis infection is now the commonest bacterial, sexually transmitted infection in the UK. Uncomplicated genital chlamydial infection often does not produce symptoms in men or women, but patients may present with sequelae. New diagnostic techniques are available to identify C. trachomatis more easily and new antibiotic regimens provide effective treatment. These factors, together with heightened awareness within the medical profession and the general population, should help to reduce the prevalence of this infection. PMID- 8730561 TI - Modern diagnosis and management of gonorrhoea. AB - Gonorrhoea remains a major global cause of morbidity. Increasingly asymptomatic infection in both women and men is recognized. Diagnosis can be via direct and indirect methods and, despite the development of newer techniques, culture remains the gold standard. The success of antimicrobial treatment depends to some extent upon the site of infection but is particularly influenced by increasing rates of antibiotic resistance. PMID- 8730562 TI - Modern diagnosis and management of acquired syphilis. AB - Syphilis presents to clinicians in a wide range of disciplines and requires a high index of suspicion. This article summarizes the interpretation of syphilis serology and provides practical guidelines for the management of acquired syphilis in liaison with local genitourinary medicine physicians. PMID- 8730563 TI - Obstetric thromboprophylaxis. AB - Thromboembolism in pregnancy is a major cause of maternal mortality. Pregnancy, particularly caesarean section, increases the risk of thrombosis. This article discusses the risk factors, the drugs used and the indications for prophylaxis of thromboembolism in pregnancy. PMID- 8730564 TI - Neurocardiogenic syncope. PMID- 8730565 TI - The leaking prosthetic valve. PMID- 8730566 TI - Management of hypertrophic cardiomyopathy. PMID- 8730567 TI - Pregnancy and heart disease. PMID- 8730568 TI - Male genital self-mutilation. AB - Male genital self-mutilation is a phenomenon which transcends nosological boundaries. This article reviews 98 cases reported in the English language literature. We note that outcome is often better than is assumed. We emphasize the importance of close liaison between the psychiatric and surgical team in managing these patients. PMID- 8730569 TI - Revision of failed total hip replacement. AB - The treatment of the patient with a failed total hip replacement has become an ever increasing problem. The incidence and aetiology of failure, and its clinical presentation, are reviewed. The planning and performance of complex revision surgery, including the use of modern bone grafting techniques and specialist implants, are outlined. Postoperative rehabilitation, current research and predictions for the future are also discussed. PMID- 8730570 TI - High-dose chemotherapy with stem cell rescue for solid tumours. PMID- 8730571 TI - Modified haemoglobins as oxygen-transporting blood substitutes. AB - Haemoglobin solutions have been developed as red cell substitutes from various sources, e.g. outdated human blood, and bovine and recombinant haemoglobins. Haemoglobins have been modified to reduce their high oxygen affinity, oncotic activity and to prolong their intravascular half-life. Aside from oxygen carriage, modified haemoglobin solutions have other interesting properties, including vasopressor effects and improvements in regional blood flow. PMID- 8730572 TI - The role of market testing in the NHS. AB - The myth of market testing is that it leads to low prices and correspondingly low quality services. If so, why have trust managers extended the initiative from the three services that are compulsory to market test, to well over 80 services? Could it be that it offers a practical and proven way to determine the best way of meeting service needs? Is it a process clinicians should fear, or is it one where clinicians have a useful and positive role to play? PMID- 8730573 TI - Immunotherapy in organ transplantation. PMID- 8730574 TI - Cutaneous lymphoma. PMID- 8730575 TI - Growth kinetics of Saccharomyces cerevisiae in batch and fed-batch cultivation using sugarcane molasses and glucose syrup from cassava starch. AB - Growth kinetics of Saccharomyces cerevisiae in glucose syrup from cassava starch and sugarcane molasses were studied using batch and fed-batch cultivation. The optimum temperature and pH required for growth were 30 degrees C and pH 5.5, respectively. In batch culture the productivity and overall cell yield were 0.31 g L-1 h-1 and 0.23 g cells g-1 sugar, respectively, on glucose syrup and 0.22 g L 1 h-1 and 0.18 g cells g-1 sugar, respectively, on molasses. In fed-batch cultivation, a productivity of 3.12 g L-1 h-1 and an overall cell yield of 0.52 g cells g-1 sugar were achieved in glucose syrup cultivation and a productivity of 2.33 g L-1 h-1 and an overall cell yield of 0.46 g cells g-1 sugar were achieved in molasses cultivation by controlling the reducing sugar concentration at its optimum level obtained from the fermentation model. By using an on-line ethanol sensor combined with a porous Teflon tubing method in automating the feeding of substrate in the fed-batch culture, a productivity of 2.15 g L-1 h-1 with a yield of 0.47 g cells g-1 sugar was achieved using glucose syrup as substrate when ethanol concentration was kept at a constant level by automatic control. PMID- 8730576 TI - Production of an extracellular polysaccharide by Agrobacterium sp DS3 NRRL B 14297 isolated from soil. AB - A bacterium isolated from soil and identified as Agrobacterium sp produced a water-soluble extracellular polysaccharide capable of producing highly viscous solutions. Gas chromatographic analysis revealed a sugar composition of glucose, galactose and mannose in the molar ratio of 7.5:2.4:1, together with 3.7% (w/w) pyruvic acid. Methylation analyses showed the presence of (1-->3)-, (1-->4)- and (1-->6)-linked glucose, (1-->3)- and (1-->4, 1-->6)-linked galactose and a small portion of (1-->3)-linked mannose residues. Succinic acid was not present. The molecular weight of the polysaccharide was estimated by light scattering to be 2 x 10(6) Da. The viscosity of solutions containing the polysaccharide remained constant from pH 3 to 11, and decreased by 50% when heated from 5 to 55 degrees C. Maximum yield of the polysaccharide, 20 g L-1, was reached in 48 h at 30 degrees C incubation. PMID- 8730577 TI - Cyclic peptides and depsipeptides from cyanobacteria: a review. AB - An elaborate array of structurally-novel and biologically-active cyclic peptides and depsipeptides are found in blue-green algae (cyanobacteria). Several of these compounds possess structures that are similar to those of natural products from marine invertebrates. Most of these cyclic peptides and depsipeptides, such as the microcystins and the lyngbyatoxins, will probably only be useful as biochemical research tools. A few, however, have the potential for development into useful commercial products. For example, cryptophycin-1, a novel inhibitor of microtubule assembly from Nostoc sp GSV 224, shows impressive activity against a broad spectrum of solid tumors implanted in mice, including multidrug-resistant ones, and majusculamide C, a microfilament-depolymerizing agent from Lyngbya majuscula, shows potent fungicidal activity and may have use in the treatment of resistant fungal-induced diseases of domestic plants and agricultural crops. PMID- 8730578 TI - The effect of internal sodium and caesium on phasic contraction of patch-clamped rabbit ventricular myocytes. AB - 1. The voltage dependence of phasic contraction was assessed in rabbit ventricular myocytes. Phasic contraction at all potentials was abolished by exposure to ryanodine-thapsigargin, showing that it was due primarily to Ca2+ release from the sarcoplasmic reticulum (SR). Experiments were performed at 35 degrees C, cells were whole-cell patch clamped and contraction was measured optically as unloaded shortening. Cells were held at -40 mV to inactivate the Na+ current (INa) and T-type Ca2+ current. A standard cellular Ca2+ load was established by applying a train of conditioning pulses at 0.5 Hz before each test pulse. The effect of replacing K+ with Cs+ in the dialysing pipette solution, and the effect of altering dialysing [Na+] between 0 and 20 mM, was assessed on contraction. 2. Cells dialysed with a K(+)-based, Na(+)-free solution exhibited a 'bell-shaped' voltage dependence of the L-type Ca2+ channel current (ICa,L), with a maximum ICa,L at +10 mV. Replacing internal K+ with Cs+, or altering pipette [Na+], did not affect the voltage dependence of ICa,L. 3. The voltage dependence of phasic contraction in cells dialysed with a K(+)-based solution was modulated by pipette [Na+]. The voltage dependence of phasic contraction was bell-shaped with 0 Na+, became much loss bell-shaped with 10 mM Na+ and with 20 mM Na+ the phasic contraction elicited at +100 mV was 1.6-fold larger than that at +10 mV. 4. Replacing 80% of K+ with Cs+ in the pipette dialysis solution led to a significant reduction in contraction amplitude and a more rapid decline in contraction amplitude after beginning the dialysis of the cell. 5. Cells dialysed with a Cs(+)-based solution displayed a voltage dependence of phasic contraction which was more bell-shaped (i.e. more similar to that of ICa,L) than that obtained with the corresponding K(+)-based dialysis solution. The level of pipette [Na+] still modulated the voltage dependence of phasic contraction in cells dialysed with a Cs(+)-based solution. 6. Time-to-peak contraction (tpk) also displayed voltage dependence; it had a minimum value between 0 and +20 mV (the voltage range for maximum ICa,L), but increased at more negative and positive potentials. Alteration of tpk contraction is discussed in relation to the stochastic behaviour of L-type Ca2+ channels and SR Ca2+ release channels. 7. The shape of the voltage dependence of contraction in rabbit myocytes at 35 degrees C is modulated by dialysing [Na+] over the tested range, 0-20 mM. Modulation of voltage dependence of contraction by dialysing [Na+] is consistent with an influence of reverse Na(+)-Ca2+ exchange in triggering intracellular Ca2+ release, in addition to the trigger Ca2+ which enters via ICa,L. 8. The marked effect of dialysing Cs+ on contraction amplitude, and on the voltage dependence of phasic contraction, does not appear to have been reported previously. Internal dialysis with Cs+ is a commonly used technique for blocking interfering outward K+ currents, in order to measure ICa,L more selectively. The present study suggests that Cs+ might also interfere with processes involved in excitation contraction coupling and indicates that it might be wise to exercise caution with the use of internal Cs+ in experiments investigating excitation-contraction coupling. PMID- 8730580 TI - Submicroscopic calcium signals as fundamental events of excitation--contraction coupling in guinea-pig cardiac myocytes. AB - 1. Subcellularly localized Ca2+ signals have been proposed to represent elementary events of cardiac Ca2+ signalling (Ca2+ sparks), whereby an individual sarcolemmal L-type Ca2+ channel locally controls opening of a single (or a few) Ca2+ release channels in the sarcoplasmic reticulum (SR). 2. To investigate directly the elementary nature of this Ca(2+)-induced Ca2+ release mechanism we used flash photolysis of caged Ca2+ while simultaneously measuring the intracellular Ca2+ concentration ([Ca2+]i) with a laser-scanning confocal microscope. 3. Power spectral analysis of the confocal images performed in the spatial domain revealed that only Ca2+ signalling events involving the L-type Ca2+ channel pathway gave rise to Ca2+ sparks. In contrast, SR Ca2+ release triggered by photolytic [Ca2+]i jumps resulted in Ca2+ transients that were always spatially homogeneous. 4. From these findings we conclude that the fundamental event of Ca2+ signalling in cardiac muscle may be smaller in size or amplitude than a Ca2+ spark. 5. We term this event a 'Ca2+ quark' possibly resulting from gating of a single SR Ca2+ release channel. It is proposed that concerted activation of several 'Ca2+ quarks' may be required for a Ca2+ spark. The 'Ca2+ quark' could also be the fundamental event in other cell types implementing a hierarchical Ca2+ signalling concept. PMID- 8730579 TI - A TTX-sensitive inward sodium current contributes to spontaneous activity in newborn rabbit sino-atrial node cells. AB - 1. Single cells were isolated from the sinus node region of rabbits (2 days old to adult) to study the age-dependent contribution of the sodium current (iNa) to pacemaker activity. 2. Experiments were conducted in 50 mM Na(+)-Ca(2+)-free solution. All newborn cells (2-19 days) exhibited a TTX-sensitive, Mn(2+) insensitive fast inward Na+ current (peak current density 115.5 +/- 11.9 pA pF-1 at 0 mV). Fifty per cent of young cells (20-40 days) possessed the current, but only one in ten adult cells. Current density decreased with development independently of cell capacitance. 3. Newborn cells exhibited a noticeable window current. With development, the position of the activation curve was shifted in the positive direction, while the inactivation was unaltered, resulting in reduced overlap of the two curves and hence less window current. 4. In newborn cells, 3 microM TTX significantly reduced all measured parameters of spontaneous action potentials, slowing rate by 63%. In contrast, there was no significant effect of TTX on rate or most of the same parameters in adult cells. 5. These results indicate that cells of the sinus node region exhibit a substantial TTX sensitive current at birth. With development, both the density and frequency of occurrence of this current within the sinus node decrease, as does its contribution to automaticity. PMID- 8730581 TI - Openings in frog microvascular endothelium induced by high intravascular pressures. AB - 1. We have investigated the effects of microvascular pressures, high enough to rupture the vessel wall, upon the ultrastructure and permeability of mesenteric capillaries in pithed frogs. 2. The vessels were perfused via micropipettes with Ringer solutions containing bovine serum albumin (1 mg ml-1) and a few human red cells. After a perfused section of vessel had been closed downstream, pressure (applied via the micropipette) was raised in a series of steps of 10 mmHg, each lasting approximately 10 s, until the vessel ruptured. Fluid filtration through the vessel wall prior to rupture was estimated from the movements of the red cells. 3. Seven vessels were fixed in glutaraldehyde immediately after rupture and prepared for electron microscopy. The electron micrographs revealed openings in the vessel walls and thirty-six of these gaps were completely defined in runs of serial sections made on four of the vessels. Twenty-nine of these gaps passed through the endothelial cells (transcellular) and seven were intercellular. 4. The pressure at which a vessel ruptured, Pb, was measured in twenty-nine vessels and had a mean +/- S.E.M. value of 79.6 +/- 5.0 cmH2O. In ten of these vessels, which had a mean +/- S.E.M. Pb of 84.2 +/- 6.5 cmH2O, microvascular pressure was lowered immediately after the initial rupture and the vessel perfused at a pressure of 20 cmH2O. Pb was then remeasured and found to be 69.9 +/- 8.4 cmH2O, which was not significantly different from its initial value. 5. Hydraulic permeability (Lp) was measured in six vessels over the range of 15-30 cmH2O before and 10 min after the vessel wall ruptured at high pressure. Mean values were 5.5 x 10(-7) and 4.0 x 10(-7) cms-1 cmH2O-1 and were not significantly different. 6. At pressures equal to and 10 mmHg below Pb, small short-lived increases in filtration rate were observed. It is suggested that these may correspond to the increased permeability to fluid and macromolecules observed at high microvascular pressures in intact capillary beds. PMID- 8730582 TI - Calcium-activated potassium channels in the endothelium of intact rat aorta. AB - 1. Single K+ channel currents and membrane potential were recorded in the endothelium of excised intact rat aorta. 2. Two types of K+ channel were found in excised patches, KCh and KAp. With Na+ and K+ as the main external and internal cations, outward conductances were 6.7 pS (KCh) and 2.8 pS (KAp). In symmetric 150 mM K+, the inward conductances were 18 and 9.1 pS. 3. Activation by Ca2+ was concentration dependent. KCh channels were activated by [Ca2+] > 0.1 microM and KAp by [Ca2+] > 0.5 microM. 4. Apamin at concentrations > 1 nM inhibited KAp Channels. Block was complete at 10 nM. KAp channels were insensitive to charybdotoxin. KCh channels were inhibited by charybdotoxin at concentrations > 50 nM, but were insensitive to apamin. 5. d-Tubocurarine (dTC) evoked flickering activity of KAp channels at concentrations > 5 microM and complete block at 100 microM. At these doses, dTC did not affect KCh channels, but at concentrations > 1 mM it decreased the single channel amplitude. 6. Hyperpolarization evoked by acetylcholine was unaffected by apamin or dTC at low concentrations ( < or = 100 microM), but inhibited by high concentrations of charybdotoxin ( > 50 nM) or dTC ( > 1 mM). 7. These data suggest that KCh channels are novel Ca(2+)-activated K+ channels responsible for the ACh-evoked hyperpolarization in the endothelium of rat aorta. PMID- 8730583 TI - Depolarization-evoked increases in cytosolic calcium concentration in isolated smooth muscle cells of rat portal vein. AB - 1. Ca2+ current through voltage-dependent Ca2+ channels (ICa) and intracellular free Ca2+ concentration ([Ca2+]i) were measured simultaneously in rat portal vein smooth muscle cells using conventional whole-cell voltage clamp technique and high temporal resolution microfluorimetry. 2. The relationship between depolarization-evoked ICa and rise in [Ca2+]i was examined. The extracellular Ca2+ concentration dependence and the voltage dependence of the depolarization evoked increases in ICa and [Ca2+]i were similar. Both ICa and increased [Ca2+]i were blocked to a similar extent by nimodipine and cadmium and augmented by Bay K 8644. Furthermore, the time course of the measured increase in [Ca2+]i, closely followed the increase in [Ca2+]i expected from the time-integrated ICa. These observations suggest that the depolarization-evoked rise in [Ca2+]i was tightly coupled to ICa. 3. The cytosolic Ca2+ buffering capacity, determined as the ratio of the expected increase in [Ca2+]i (from ICa) divided by the measured increase in [Ca2+]i, was over 100. Therefore, less than 1 out of 100 Ca2+ ions entering the cell appears as a free Ca2+. 4. Ryanodine (30 microM), a blocker of the Ca(2+)-induced Ca2+ release mechanism, had little effect on buffering capacity measured over the first 200 ms of the depolarizing voltage clamp pulse. Ryanodine also had little effect on the buffering capacity during 800-1000 ms of the depolarizing voltage clamp pulse. Therefore, it was concluded that there is little Ca(2+)-induced Ca2+ release from the stores in rat portal vein smooth muscle cells during depolarization-evoked Ca2+ entry. 5. During brief depolarizations, the largest [Ca2+]i increase and ICa occurred at 0 mV. However, during steady-state depolarization, the largest increase in [Ca2+]i occurred around -30 mV, and we estimate the peak steady-state ICa to be about 0.6 pA. PMID- 8730584 TI - Dual effect of external Ca2+ on noradrenaline-activated cation current in rabbit portal vein smooth muscle cells. AB - 1. The influence of divalent cations was investigated on the noradrenaline activated non-selective cation current (Icat) in freshly dispersed smooth muscle cells of rabbit portal vein. 2. Reduction of external calcium concentration ([Ca2+]o) from 1.5 mM to 'test' levels of 100-500 microM during stimulation of Icat produced a sustained potentiation of Icat, which indicates an inhibitory action of Cao2+. With 'test' concentrations lower than 100 microM Cao2+ the amplitude of Icat was initially potentiated and then the current subsequently declined. Under these conditions re-addition of 1.5 mM Cao2+ transiently activated Icat which demonstrates an additional facilitatory action of [Ca2+]o. The half-maximal [Ca2+]o for the facilitatory and inhibitory action on Icat was 6 and 400 microM, respectively. 3. When [Ca2+]o was decreased to very low levels (< 10 nM) by inclusion of 1 mM BAPTA in nominally Ca(2+)-free external solution, Icat could still be evoked by noradrenaline. This suggested that Ca2+ is not obligatory for activation of Icat but merely modulatory. 4. In 1.5 mM Cao2+ the current-voltage (I-V) relationship of Icat was S-shaped with a reversal potential (Er) of about +9 mV. The I-V characteristics and Er of the potentiated current in 200 microM Cao2+ were not markedly different from those in 1.5 mM Cao2+. 5. During the noradrenaline-induced Icat, removal of external Mg2+ caused a small increase in the amplitude of Icat which was not as great as that seen on reduction of [Ca2+]o and there was no facilitatory effect of Mgo2+. 6. These results indicate that external Ca2+ has a dual action on Icat, both facilitating and inhibiting the current. This dual action is not shared with external Mg2+. PMID- 8730585 TI - Enhancement of ionic current and charge movement by coexpression of calcium channel beta 1A subunit with alpha 1C subunit in a human embryonic kidney cell line. AB - 1. Coexpression of the beta subunit with the alpha 1C subunit of the cardiac L type Ca2+ channel has been shown to increase ionic current. To examine the mechanism of this increase, ionic and gating currents were measured in transiently transfected HEK293 cells. 2. Beta 1A subunit coexpression increased the maximal whole-cell conductance (Gmax) measured in 10 mM Ba2+ from 91 +/- 11 to 833 +/- 107 pS pF-1 without a change in the voltage dependence of activation (V1/2: -6.1 +/- 1.1 and -6.6 +/- 0.9 mV, respectively). 3. Gating currents were smaller in cells expressing only the alpha 1C subunit (only four out of eleven cells exhibited gating currents above the limits of detection, whereas eight out of eight beta 1A coexpressing cells had measurable gating currents). The gating currents were integrated to measure the intramembrane charge movement (Q). The ON charge movement (Qon) could be described by a Boltzmann distribution reaching a maximal value of Qon,max. 4. The mean ratio of Gmax: Qon,max increased from 99 +/ 6 to 243 +/- 30 pS fC-1 with beta 1A coexpression, demonstrating that the beta 1A subunit changes the gating of alpha 1C channels to favour the opening of the channels. However, this 2.5-fold change in the Gmax: Qon,max ratio explains less than half of the 9.2-fold increase in Gmax with beta 1A subunit coexpression. The major effect is due to a 3.7-fold increase in Qon,max, demonstrating that beta 1A subunit coexpression increases the number of functional surface membrane channels. PMID- 8730587 TI - Activation and blockade of mouse muscle nicotinic channels by antibodies directed against the binding site of the acetylcholine receptor. AB - 1. Using the patch-clamp technique, we have found that mouse muscle nicotinic acetylcholine receptor (nAChR) channels can be activated by low concentrations of a monoclonal antibody (MoAb), referred to as WF6, which is directed against the acetylcholine (ACh) binding site. Similar effects were seen using IgG or F(ab)2 fragments from the sera of patients with myasthenia gravis (MG), which contain polyclonal anti-nAChR antibodies. 2. The mean open times of MoAb and the slope conductance of single WF6-activated single channels were similar to those of ACh activated channels under the same experimental conditions. 3. On outside-out patches, single channel activity was elicited by MoAb WF6 and MG F(ab)2 fragments, and was blocked by (+)-tubocurarine. We therefore concluded that MoAb WF6 and the MG F(ab)2 fragments activate the nAChR. 4. MoAb WF6 and MG F(ab)2 fragments blocked the current activated by pulsed application of 10(-4) M ACh to a significant extent. The block was partly reversible. The rate constants for the binding and dissociation of MoAb WF6 from the receptor were determined quantitatively. PMID- 8730586 TI - Modulation of the hyperpolarization-activated current (Ih) by cyclic nucleotides in guinea-pig primary afferent neurons. AB - 1. Whole-cell patch-clamp recordings were made from dissociated guinea-pig nodose and trigeminal ganglion neurons in culture to study second messenger mechanisms of the hyperpolarization-activated current (Ih) modulation. 2. Prostaglandin E2 (PGE2) and forskolin modulate Ih in primary afferents by shifting the activation curve in the depolarizing direction and increasing the maximum amplitude. 3. The cAMP analogues, RP-cAMP-S (an inhibitor of protein kinase A (PKA)) and SP-cAMP-S (an activator of PKA), both shifted the activation curve of Ih to more depolarized potentials and occluded the effects of forskolin. These results suggest that Ih is modulated by a direct action of the cAMP analogues. 4. Superfusion of other cyclic nucleotide analogues (8-Br-cAMP, 8-(4 chlorophenylthio)-cAMP and 8-Br-cGMP) mimicked the actions of forskolin and PGE2, but dibutyryl cGMP, 5'-AMP and adenosine had no effect on Ih. 8-Br-cAMP and 8-Br cGMP had similar concentration response profiles, suggesting that Ih has little nucleotide selectivity. 5. The inhibitor peptide (PKI), the catalytic subunit of PKA (C subunit) and phosphatase inhibitors (microcystin and okadaic acid) had no effect on forskolin modulation of Ih. 6. These results indicate that Ih is regulated by cyclic nucleotides in sensory neurons. Positive regulation of Ih by prostaglandins produced during inflammation may lead to depolarization and facilitation of repetitive activity, and thus contribute to sensitization to painful stimuli. PMID- 8730588 TI - High-threshold Ca2+ currents in rat hippocampal interneurones and their selective inhibition by activation of GABA(B) receptors. AB - 1. Whole-cell calcium currents were recorded from visually identified inhibitory interneurones located in stratum radiatum (near the border with stratum lacunosum moleculare) of area. CA1 in rat hippocampal slices. Current-voltage (I-V) relationships in relatively well-clamped neurones showed that inward current activated between -50 and -40 mV (holding potential, -80 mV) and was maximal near -10 mV. Currents showed little inactivation over the course of 85 ms steps, and were completely blocked by removal of Ca2+ or addition of Cd2+. Prominent low threshold currents were not observed under these conditions. 2. The calcium channels contributing to whole-cell currents in interneurones were examined using selective channel antagonists. The selective N-type calcium channel blocker omega conotoxin GVIA (omega-CgTX-GVIA; 10 microM) irreversibly blocked 23.2 +/- 2.8% of whole-cell currents. The P/Q-type antagonist omega-agatoxin IVA (omega-Aga-IVA; 1 5 microM) blocked 10.4 +/- 3.3% of whole-cell currents. Block by omega-Aga-IVA was highly variable, ranging from 0 to 30%. The less selective conotoxin, omega conotoxin MVIIC (omega-CTX-MVIIC; 5 microM) blocked 31.0 +/- 2.7% of whole-cell currents. The selective L-type channel antagonist nifedipine (20 microM) blocked 27.5 +/- 3.5% of whole-cell currents. 3. Whole-cell calcium currents were reversibly inhibited by the selective GABA(B) receptor agonists (+/-)-baclofen or CGP 27492 (1-3 microM; 18.9 +/- 1.4%). This inhibition was reversed or prevented by the selective GABAB receptor antagonist CGP 55845A (1 microM). Inhibition of inward current activated by voltage ramps was voltage dependent, being greatest near -10 mV, and less pronounced at more positive or negative potentials. Inhibition of calcium currents by GABAB receptor agonists was accompanied by an apparent change in the kinetics of whole-cell currents consistent with a slowing of the rate of activation. CGP 27492 depressed calcium currents by 16.1 +/- 1.9% before application of omega-CgTX-GVIA, and by 3.9 +/- 2.0% after application of omega-CgTX-GVIA in the same cells (P < 0.005), consistent with preferential block of N-type calcium channels. 4. Neither adenosine (200 microM) nor the selective mu-opioid receptor agonist Tyr-D-Ala-Gly-MePhe-Gly-ol (DAMGO; 2 microM) inhibited calcium currents. Similarly, CGP 27492, but not adenosine or DAMGO, induced an outward current (at - 70 mV) consistent with activation of inwardly rectifying potassium channels. 5. These results indicate that hippocampal inhibitory neurones located in stratum radiatum possess multiple calcium channel subtypes, including N-type, L-type, and at least two other types of high-threshold channels. Activation of GABAB receptors (but not adenosine or mu-opioid receptors) preferentially inhibits N-type channels in these neurones. Similar inhibition occurring in the terminals of interneurones could contribute to depression of inhibitory synaptic transmission by activation of GABAB autoreceptors. PMID- 8730589 TI - Effect of RNA editing and subunit co-assembly single-channel properties of recombinant kainate receptors. AB - 1. Patch-clamp methods have been used to examine single-channel properties of recombinant GluR5 and GluR6 kainate-preferring glutamate receptors which differ in a single amino acid residue as a result of RNA editing at the Q/R (glutamine/arginine) site. Subunits were expressed alone or in combination with the high-affinity kainate receptor subunit KA - 2 in transfected human embryonic kidney (HEK-293) cells. 2. In outside-out patches, unedited homomeric GluR6(Q) receptors exhibited directly resolved domoate-activated single-channel conductances of 8, 15 and 25 pS. Variance analysis of GluR6(Q) responses gave a mean conductance of 5.4 pS, while the edited isoform GluR6(R) had an unusually low channel conductance (225 fS). 3. Homomeric channels composed of GluR5(Q) subunits exhibited three conductance states of 5, 9 and 14 pS characterized by prolonged burst activations in the presence of domoate. In contrast, the GluR5(R) subunit, which has not previously been reported to form functional homomeric receptors, had an extremely low conductance (< 200 fS). 4. Heteromeric GluR6(Q)/KA-2 kainate receptors gave single-channel events indistinguishible from homomeric GluR6(Q) channels. Conversely, openings produced by GluR5(Q)KA-2 and GluR5(Q) receptors differed from each other in their kinetic properties. The primary effect of co-expression of KA-2 with GluR5(Q) was a dramatic shortening in channel burst length. 5. Spectral and variance analyses were used to estimate mean single-channel conductances of heteromeric edited receptor-channels; channel conductances were 950 fS for GluR5(R)KA-2 receptors and 700 fS for GluR6(R)/KA-2 receptors. Both receptor types had significantly higher conductances than the respective homomeric channels, GluR5(R) and GluR6(R). 6. We conclude that Q/R site editing dramatically reduces single-channel conductance. Furthermore, we find similarity between the kainate receptor-channels described in sensory neurones and the recombinant GluR5(Q) homomeric channel. Characterization of recombinant single-channel properties could therefore aid identification of the native kainate receptors. PMID- 8730590 TI - Endogenous and exogenous dopamine depress EPSCs in rat nucleus accumbens in vitro via D1 receptors activation. AB - 1. Whole-cell patch clamp recordings were made from nucleus accumbens neurones in slices of rat ventral forebrain. In the presence of picrotoxin (50 microM), the amplitude of 6-cyano-7-nitroquinoxaline-2, 3-dione-sensitive glutamate EPSCs, recorded at holding potentials between -80 and -90 mV, was reversibly reduced by 56 +/- 11% (n = 6) by dopamine (30 microM). The selective dopamine D1 receptor agonists SKF 38393 (10 microM) and SKF 81297A (10 microM), but not the selective D2 receptor agonist quinpirole (10 microM), also reversibly depressed the EPSC by 36-48%. The depression of the EPSC by dopamine was completely blocked by the D1 receptor antagonist SCH 23390 (1 microM), whereas the D2 antagonist (-)-sulpiride (1 microM) was without effect. 2. EPSCs were reversibly depressed by the dopamine mimetic psychostimulants cocaine (1-20 microM) and amphetamine (10-30 microM) by 40 +/- 16 and 62 +/- 9%, respectively, but only in about half of the cells tested (11/23 and 6/13, respectively). Their actions were completely reversed by SCH 23390 (1 microM), indicating that endogenous dopamine can also depress the EPSC via D1 receptors. 3. No discernable effects of dopamine, SKF 81297A, SKF 38393, quinpirole, cocaine or amphetamine were observed on membrane conductance or holding current (at holding potentials of -80 to -90 mV), suggesting that the depression of the EPSC was solely due to an action on presynaptic D1 receptors. 4. In contrast, agents that elevate intracellular levels of adenosine-3':5' cyclic monophosphate (cAMP) (forskolin (1-10 microM), 3-isobutyl-1-methylxanthine (0.1-1 mM), rolipram (10 microM), and dibutyryl cAMP (0.5-1 mM)) caused a reversible increase in the EPSC amplitude (by 21-150%). Furthermore, in the presence of forskolin (10 microM), the ability of dopamine to depress synaptic transmission was unaffected. 5. Together these data suggest that both exogenous dopamine and dopamine released from intrinsic nerve terminals attenuate glutamate receptor-mediated synaptic transmission in the nucleus accumbens by presynaptic D1 receptor activation. The transduction mechanism underlying this effect does not appear to involve the formation of cAMP. PMID- 8730591 TI - Spontaneous and uniquantal-evoked endplate currents in normal frogs are indistinguishable. AB - 1. A recent paper concludes that the shapes of spontaneous and uniquantal-evoked signals are different. The signals were recorded with extracellular electrodes, often in the presence of neostigmine. Differences were reported between the voltage-time integrals, and between the decay times of spontaneous and evoked signals. 2. These results disagree with earlier studies using the two-electrode voltage clamp technique. 3. We recorded miniature-endplate currents (MEPCs) and uniquantal-endplate currents (u-EPCs) in a low-Mg(2+)-Ca2+ solution, sometimes with neostigmine present. Evoked quantal outputs were estimated by the method of failures, so we could reject the appropriate number of the largest evoked releases. The twenty-nine experiments showed that there were no consistent differences between the current-time integrals or half-decay times (t1/2), regardless of whether or not neostigmine was present. 4. When recording simultaneously with intracellular and extracellular electrodes, on average about 25% of the miniatures were seen in both recordings. On average, 63% of the endplate potentials were also seen in both recordings. Extracellular recording may not give the precise localization generally assumed. 5. We again conclude that quanta released by nerve stimulation and spontaneously are indistinguishable at normal frog neuromuscular junctions. PMID- 8730592 TI - Effects of troponin C isoforms on pH sensitivity of contraction in mammalian fast and slow skeletal muscle fibres. AB - 1. The effects of troponin C (TnC) isoforms on the acidic pH-induced rightward shift in the tension-pCa (-log[Ca2+]) relationship were examined in slow soleus and fast psoas skeletal muscle fibers. Endogenous TnC was partially extracted from skinned single fibres and the extracted fibres were subsequently reconstituted with purified TnC. The pCa producing one-half maximal tension (pCa50) was determined at pH 7.00 and 6.20 in each fibre and then the pH-induced shift in pCa50 (delta pCa50) was calculated. 2. In control fast fibres which express fast skeletal TnC (sTnC), the delta pCa50 was 0.64 +/- 0.02 pCa units (n = 10), and this increased significantly to 0.78 +/- 0.04 pCa units (n = 8) following extraction and reconstitution with cardiac TnC (cTnC). In each fibre, the reconstituted delta pCa50 was subtracted from the control delta pCa50 which yielded a significant shift of -0.13 +/- 0.05 pCa units (n = 8; P < 0.05). Thus, the pH sensitivity of contraction was increased in the cTnC-reconstituted psoas fibres. 3. In extracted psoas fibres that were reconstituted with fast sTnC the pH sensitivity of contraction was unchanged, indicating that the above effects were related to the TnC isoform and not a non-specific effect of the extraction procedure. 4. In a second series of experiments cTnC was specifically extracted from slow soleus fibres which were subsequently reconstituted with purified fast sTnC. Skeletal TnC reconstituted soleus fibres demonstrated a significant decrease in pH sensitivity. In each fibre, the reconstituted delta pCa50 (mean, 0.58 +/- 0.02 pCa units) was subtracted from the control delta pCa50 (mean, 0.63 +/- 0.02 pCa units) which yielded a significant shift of 0.05 +/- 0.01 pCa units (n = 4; P < 0.05). The pH sensitivity was not altered in cTnC-reconstituted soleus fibres (-0.01 +/- 0.01 pCa units, n = 4). 5. These findings indicate that TnC isoforms alter the pH sensitivities of contraction in slow and fast skeletal muscle fibres. However, the magnitude of the change in pH sensitivity is muscle lineage dependent, indicating that differential expression of other myofilament protein isoforms, together with TnC, is necessary to confer full pH sensitivity of contraction in striated muscles. PMID- 8730593 TI - Selective depletion of spinal monoamines changes the rat soleus EMG from a tonic to a more phasic pattern. AB - 1. To assess the role of descending monoaminergic pathways for motor activity long-lasting EMG recordings were performed from the adult soleus muscle before and after selective depletion of spinal monoamines. 2. Rats were chronically implanted with an intrathecal catheter placed in the lumbar subarachnoid space and gross-EMG recording electrodes in the soleus muscle. EMG recordings were performed in control conditions and at different times after intrathecal administration of either 40-55 micrograms 5,6-dihydroxytryptamine (5,6-DHT) and 40-55 micrograms 6-hydroxydopamine (6-OHDA) or 80 micrograms 5,7 dihydroxytryptamine (5,7-DHT) alone. The depletions were evaluated biochemically in brains and spinal cords after recordings. 3. In agreement with previous studies the intrathecal administration of neurotoxins caused a reduction of the noradrenaline (NA) and serotonin (5-HT) content of the lumbar spinal cord to about 2-3% of control, with little or no changes in the monoamine content of the cortex. 4. In non-treated chronically catheterized rats the integrated rectified gross EMG displayed long-lasting EMG episodes composed of phasic high-amplitude events and tonic segments of varying duration and amplitude. 5. After intrathecal administration of neurotoxins the number of long-lasting gross-EMG episodes, the mean episode duration, and the total EMG activity per 24 h, were reduced. These changes were accompanied by a simultaneous increase both in the number of short lasting EMG episodes and the total number of EMG episodes per 24 h period. The changes were apparent 5-6 days after drug administration and fully developed after 2-3 weeks. 6. No changes in general movement ability were observed, except that the denervated animals had a tendency to a less errect posture. 7. These results indicate that descending monoaminergic pathways are important for the maintained motor output in tonic hindlimb muscles. PMID- 8730594 TI - Electrical properties of smooth muscle in the guinea-pig urinary bladder. AB - 1. The effects of transmural nerve stimulation were examined on preparations of detrusor smooth muscle from guinea-pig urinary bladder using intracellular recording techniques. Most recordings were made from preparations in which spontaneous and evoked action potentials had been inhibited by nifedipine (10 microM), a dihydropyridine that blocks L-type Ca2+ channels. 2. Supramaximal stimuli evoked excitatory junction potentials (EJPs) which could be divided into three basic types. Type 1 EJPs had short latencies (< 30 ms) and fast rise times (< 60 ms). Type 2 EJPs consisted of two components: a small depolarization that was followed by a second depolarization with a faster rise time. In a third type of cell, at high strengths of stimulation, EJPs resembled type 1 EJPs but at lower strengths of stimulation were similar in time course to type 2 EJPs. 3. All EJPs were abolished by tetrodotoxin (1 microM) and reduced by omega-conotoxin (0.1 microM), but were unaffected by hexamethonium (0.1 mM), suggesting that they result from the release of transmitter from post-ganglionic nerve fibres. All responses persisted in the presence of atropine (1 microM) but were abolished following the desensitization of P2-purinoceptors with alpha, beta-methylene ATP (m-ATP; 10 microM). 4. Spontaneous excitatory junction potentials (SEJPs) were also recorded from most cells. SEJPs were similar in appearance to fast single component EJPs; however, in general they had a briefer time course. SEJPs persisted in the presence of tetrodotoxin (1 microM). 5. The electrical properties of urinary bladder smooth muscle were also examined. Voltage changes induced by point current injection into cells had fast rates of rise and decay (time constant, 5-20 ms). The input resistance of cells ranged between 12 and 108 M omega. When recordings were taken from cells near the point of current injection, resultant electrotonic potentials could be detected in only a small proportions of cells. 6. The results are discussed in relation to the idea that transmural nerve stimulation in the guinea-pig urinary bladder causes the activation of at least two different membrane conductances. Cells appear to be electrically coupled with one another. However, it is likely that coupling exists within discrete bundles of the smooth muscle. PMID- 8730595 TI - Spike after-depolarization and burst generation in adult rat hippocampal CA1 pyramidal cells. AB - 1. Intracellular recordings in adult rat hippocampal slices were used to investigate the properties and origins of intrinsically generated bursts in the somata of CA1 pyramidal cells (PCs). The CA1 PCs were classified as either non bursters or bursters according to the firing patterns evoked by intrasomatically applied long ( > or = 100 ms) depolarizing current pulses. Non-bursters generated stimulus-graded trains of independent action potentials, whereas bursters generated clusters of three or more closely spaced spikes riding on a distinct depolarizing envelope. 2. In all PCs fast spike repolarization was incomplete and ended at a potential approximately 10 mV more positive than resting potential. Solitary spikes were followed by a distinct after-depolarizing potential (ADP) lasting 20-40 ms. The ADP in most non-bursters declined monotonically to baseline ('passive' ADP), whereas in most bursters it remained steady or even re depolarized before declining to baseline ('active' ADP). 3. Active, but not passive, ADPs were associated with an apparent increase in input conductance. They were maximal in amplitude when the spike was evoked from resting potential and were reduced by mild depolarization or hyperpolarization (+/- 2 mV). 4. Evoked and spontaneous burst firing was sensitive to small changes in membrane potential. In most cases maximal bursts were generated at resting potential and were curtailed by small depolarizations or hyperpolarizations (+/- 5 mV). 5. Bursts comprising clusters of spikelets ('d-spikes') were observed in 12% of the bursters. Some of the d-spikes attained threshold for triggering full somatic spikes. Gradually hyperpolarizing these neurones blocked somatic spikes before blocking d-spikes, suggesting that the latter are generated at more remote sites. 6. The data suggest that active ADPs and intrinsic bursts in the somata of adult CA1 PCs are generated by a slow, voltage-gated inward current. Bursts arise in neurones in which this current is sufficiently large to generate suprathreshold ADPs, and thereby initiate a regenerative process of spike recruitment and slow depolarization. PMID- 8730596 TI - Ionic basis of spike after-depolarization and burst generation in adult rat hippocampal CA1 pyramidal cells. AB - 1. Intracellular recordings in adult rat hippocampal slices were used to identify the ionic conductances underlying active spike after-depolarization (ADP) and intrinsic burst firing in the somata of CA1 pyramidal cells (PCs). To test the 'Ca2+ hypothesis', Ca2+ currents were suppressed by replacing the Ca2+ in the saline with either Mn2+ or Mg2+. Alternatively, the inorganic Ca2+ channel blockers Cd2+ (0.5 mM) or Ni2+ (2 mM) were added to the saline. To test the 'Na+ hypothesis', Na+ currents were blocked with tetrodotoxin (TTX; 0.5 microM). 2. The suppression of Ca2+ currents blocked the fast after-hyperpolarization (AHP) generated by the fast Ca(2+)-gated K+ current Ic, while enhancing the amplitude and duration of active spike ADPS. 3. Evoked and spontaneous burst firing was preserved undiminished following Ca2+ current suppression, while the propensity to fire bursts increased in many cases. The postburst medium AHP (generated primarily by the muscarine-sensitive voltage-gated K+ current, IM) was not affected by this treatment, which blocked the slow AHP (generated by the slow Ca(2+)-gated K+ current, IAHP). 4. TTX strongly suppressed active ADPs and intrinsic bursts before substantially reducing the threshold, rate of rise and amplitude of solitary spikes. 5. In Ca(2+)-free saline, caesium-filled PCs generated large, plateau ADPs following an initial burst of fast spikes. Application of TTX suppressed these ADPs before solitary fast spikes appeared to be reduced. 6. Injection of brief, just subthreshold depolarizing current pulses into bursters evoked slow depolarizing potentials lasting up to 50 ms. These persisted after suppression of Ca2+ currents and were entirely blocked by TTX. 7. We conclude that active spike ADPs and intrinsic bursts in the somata of adult CA1 PCs are generated by a low voltage-gated, persistent Na+ current. Burst termination is mediated by voltage-gated K+ currents activated during the burst (most likely IM), rather than by the Ca(2+)-gated K+ currents Ic and IAHP. The latter currents downregulate the innate tendency of CA1 PCs to burst (Ic) and limit the rate of spontaneous burst firing (IAHP). PMID- 8730597 TI - A novel synaptic interaction underlying induction of long-term depression in the area CA1 of adult rat hippocampus. AB - 1. We describe a novel synaptic property that regulates induction of homosynaptic long-term depression (LTD), and slowly developing heterosynaptic LTD, of Schaffer collateral-pyramidal cell synapses in adult rat hippocampus. 2. Two independent pathways converging on the same neuron were alternately tested with 0.017 Hz single pulses, and LTD was induced by 900 conditioning stimuli delivered at 2 Hz. All experiments were performed in the presence of the GABA(A) antagonists picrotoxin or bicuculline. 3. After delivery of the 2 Hz stimulation to the homosynaptic pathway, the 0.017 Hz test pulses to the heterosynaptic pathway were interrupted for 25 min. When the test stimulations were resumed, heterosynaptic LTD could not be observed. Homosynaptic LTD also failed to be induced in this protocol. Interruption of test pulses did not itself cause a general increase of synaptic responses. 4. Doubling the frequency of homosynaptic test pulses (to 0.033 Hz) during a 25 min interruption of heterosynaptic stimulus did not preserve homosynaptic LTD. This suggests that the failure of homosynaptic LTD induction seen when the test pulses were interrupted was not caused by a decrease in the number of synaptic inputs at the postsynaptic neuron following conditioning. 5. When only the homosynaptic pathway was involved, with no heterosynaptic stimulation, as in conventional experiments, 2 Hz conditioning successfully induced homosynaptic LTD. 6. We propose that when a heterosynaptic pathway has been recently used, continuous input to that pathway following conditioning is necessary for induction of homosynaptic LTD on the same neuron. PMID- 8730598 TI - Cellular mechanisms for neuronal thermosensitivity in the rat hypothalamus. AB - 1. To study the basic mechanisms of neuronal thermosensitivity, rat hypothalamic tissue slices were used to record and compare intracellular activity of temperature-sensitive and -insensitive neurones. This study tested the hypothesis that different neuronal types have thermally dependent differences in the transient potentials that determine the interspike interval. 2. Most spontaneously firing neurones displayed depolarizing prepotentials that preceded each action potential. In warm-sensitive neurones, warming increased the rate of rise of the depolarizing prepotential which, in turn, decreased the interspike interval and increased the firing rate. In contrast, temperature had little or no effect on the rate of rise in prepotentials of temperature-insensitive neurones. 3. Prepotential depolarization can be due to increasing depolarizing conductances or decreasing hyperpolarizing conductances. These are differences in the ionic conductances responsible for prepotentials in temperature-sensitive and insensitive neurones. In warm-sensitive neurones, the net ionic conductance decreased as the prepotential depolarized towards threshold, suggesting that the prepotential is primarily determined by a decrease in outward potassium conductances. In contrast, in low-slope temperature-insensitive neurones, the net conductance remained constant during the interspike interval, suggesting a more balanced combination of both depolarizing and hyperpolarizing conductances. 4. Transient outward potassium currents, including A-currents, are important determinants of neuronal firing rates. These currents were identified in all warm sensitive neurones tested, as well as in many temperature-insensitive and silent neurones. Since warming increased the rates of inactivation of these currents, transient K+ currents may contribute to the temperature-dependent prepotentials of some hypothalamic neurones. PMID- 8730599 TI - Effects of hypernatraemia in the central nervous system and its therapy in rats and rabbits. AB - 1. We studied the effects of acute (1 or 4 h) and chronic (1 week) hypernatraemia (plasma [Na+], 170-190 mM) on brain histology, and brain water and solute contents in rats and rabbits. 2. In rabbits with acute hypernatraemia, there was significant loss of intracellular brain water, with increases in brain [Na+ + K+], amino acid concentration, and undetermined solute (idiogenic osmole). After 1 week of recovery, brain intracellular water content had returned to normal. 3. In hypernatraemic rats there was myelinolysis of brain white matter, with karyorrhexis and necrosis of neurons. 4. Hypernatraemic rabbits were treated with 77 mM NaCl (i.v.) to normalize plasma [Na+] over 4-24 h intervals. Therapy of either acute or chronic hypernatraemia resulted in significant brain oedema because brain osmolality failed to decrease at the same rate as plasma osmolality. 5. It is concluded that: (a) untreated hypernatraemia results in brain lesions demonstrating myelinolysis and cellular necrosis; (b) normalization of hypernatraemia over 4-24 h results in cerebral oedema, due primarily to failure of brain amino acids and idiogenic osmoles to dissipate as plasma [Na+] is decreased to normal. PMID- 8730600 TI - Stimulation of secretion from bovine adrenal chromaffin cells by microsecond bursts of therapeutic levels of ultrasound. AB - 1. In this study the secretory response of individual bovine adrenal chromaffin cells was monitored using amperometric carbon-fibre microelectrodes. Cells were stimulated to secrete by exposure to 20-100 microseconds long tonebursts of ultrasound (2-4 x 10(5) Pa; peak pressure at 1 MHz). 2. Three types of secretory responses were observed: an almost instantaneous response, a delayed release of catecholamines, or a series of 'burst-like' secretory bouts. 3. Fura-2 measurements of intracellular Ca2+ concentrations showed that the release of catecholamines was accompanied by an increase in the intracellular Ca2+ concentration. In the absence of extracellular Ca2+, secretory responses were not evoked showing that Ca2+ entry was necessary to elicit catecholamine release. PMID- 8730601 TI - Rib motion modulates inspiratory intercostal activity in dogs. AB - 1. A test was performed of the hypothesis that the motion of the ribs during inspiration modulates, via changes in spindle afferent activity, the activation of the inspiratory intercostal muscles. The electrical activity of the parasternal intercostal, external intercostal, and levator costae muscles in anaesthetized spontaneously breathing dogs was thus recorded during manipulation of the inspiratory displacement of the ribs over a wide range of rib motion. 2. In agreement with the hypothesis, the external intercostal and levator costae muscles lengthened and showed increased inspiratory activities when the normal inspiratory cranial motion of the lower rib was reduced or reversed into an inspiratory caudal motion. Conversely, the inspiratory activities decreased when the inspiratory cranial motion of the rib and the inspiratory shortening of the muscles was augmented. The inspiratory activity of the parasternal intercostal remained unchanged throughout. 3. However, when the two ribs making up the interspace were linked together so that the external intercostal muscle was constant in length, the relationship of muscle activity to rib motion was maintained. 4. In addition, when the upper rather than the lower rib of the interspace was manipulated, the relationship between the change in muscle length and inspiratory activity was reversed, so that activity decreased when the muscle was lengthened and increased when the muscle was shortened. The relationship of muscle activity to lower rib motion, however, was still maintained. 5. These observations thus indicate that rib motion triggers proprioceptive reflexes which, regardless of the changes in length of the individual muscles, make the external intercostal inspiratory activity exquisitely sensitive to the direction of rib displacement. PMID- 8730602 TI - Chemosensitive medullary neurones in the brainstem--spinal cord preparation of the neonatal rat. AB - 1. Using the isolated medulla and spinal cord of the neonatal rat, the response to CO2-induced changes in superfusate pH was examined in whole cell and perforated patch recordings from ventral medullary neurones which were identified by injection of Lucifer Yellow. The respiratory response to changing the CO2 concentration (from 2 to 8%) consisted of an increase in phrenic burst frequency, which could be accompanied by an increase, decrease or no change in burst amplitude. 2. Five classes of neurone - inspiratory, post-inspiratory, expiratory, respiration-modulated and ionic - were distinguished on the basis of their membrane potential and discharge patterns. Almost all (112 of 123) responded rapidly to 8% CO2 with a sustained change in membrane potential. Depolarizing responses (3-18 mV) occurred in inspiratory, respiration-modulated and 45% of tonic neurones. Hyperpolarizing responses (2-19 mV) occurred in expiratory and post-inspiratory neurones. The remaining tonic neurones were inhibited or showed no response. 3. In representatives of each class of neurone, membrane potential responses to 8% CO2 were retained when tested in the presence of tetrodotoxin (n = 7), low (0.2 mM) Ca(2+)-high (5 mM) Mg2+ (n = 23) or Cd2+ (0.2 mM) (n = 3)-containing superfusate, implying that they are mediated by intrinsic membrane or cellular mechanisms. 4. Neurones were distributed between 1200 microns rostral and 400 microns caudal to obex, and their cell bodies were located between 50 and 700 microns below the ventral surface (n = 104). Almost all responsive neurones (n = 78) showed dendritic projections to within 50 microns of the surface. 6. These experiments indicate that significant numbers of ventral medullary neurones, including respiratory neurones, are intrinsically chemosensitive. The consistency with which these neurones show surface dendritic projections suggests that this sensitivity may arise in part at this level. PMID- 8730603 TI - Interaction of training and diet on metabolism and endurance during exercise in man. AB - 1. Ten untrained young men ingested a carbohydrate-rich diet (65 energy percent (E%) carbohydrate, T-CHO) and ten similar subjects a fat-rich diet (62 E% fat, T FAT) while endurance training was performed 3-4 times a week for 7 weeks. For another 8th week of training both groups ingested the carbohydrate-rich diet (T CHO and T-FAT/CHO). 2. Maximal oxygen uptake increased by 11% (P < 0.05) in both groups after 7 and 8 weeks. Time to exhaustion at 81% of pre-training maximal oxygen uptake increased significantly from a mean (+/- S.E.M.) of 35 +/- 4 min to 102 +/- 5 and 65 +/- 7 min in T-CHO and T-FAT, respectively, after 7 weeks (P < 0.05, T-CHO vs. T-FAT). After 8 weeks, endurance remained unchanged in T-CHO but increased (P < 0.05) to 77 +/- 9 min in T-FAT/CHO which, however, was still less (P < 0.05) than in T-CHO. 3. Muscle glycogen breakdown rate during exercise was halved by endurance training equally in both T-CHO and T-FAT after 7 and 8 weeks, and muscle glycogen stores at exhaustion were not depleted in any group. 4. During exercise after 7 weeks, the respiratory exchange ratio (RER) was unchanged in T-CHO (0.88 +/- 0.01) compared with pre-training but decreased (P < 0.05) to 0.82 +/- 0.02 in T-FAT. After 8 weeks, RER in both T-CHO and T-FAT/CHO was approximately 0.87. 5. During exercise, plasma noradrenaline concentration and heart rate were higher in T-FAT than in T-CHO both at 7 and at 8 weeks. 6. It is concluded that ingesting a fat-rich diet during an endurance training programme is detrimental to improvement in endurance. This is not due to a simple lack of carbohydrate fuel, but rather to suboptimal adaptations that are not remedied by short-term increased carbohydrate availability. Furthermore, the study suggests that the decrease in RER usually seen after training when exercising at the same absolute intensity as before training can be prevented by a carbohydrate-rich diet. PMID- 8730604 TI - Foveal cone mosaic and visual pigment density in dichromats. AB - 1. Optical reflectance spectra of the fovea were measured in ten subjects with normal colour vision, ten protanopes and seven deuteranopes. Four conditions were used: perpendicular and oblique angle of incident and reflected light on the retina, both in a dark-adapted and a fully bleached state. 2. The spectra were analysed to assess the effects of dichromacy on the cone mosaic. A replacement model, i.e. one where the total number of cones remains unchanged and all cones are filled with a single type of pigment, was found to fit our data best. 3. The analysis of the spectral fundus reflectance also provided estimates for densities of photo-labile and photo-stable retinal pigments and fraction of long wavelength sensitive (LWS) cones. Visual pigment density was 0.39 for protanopes and 0.42 for deuteranopes, significantly lower than the 0.57 found for colour normals. Macular pigment density was 0.54 for colour normals, 0.46 for protanopes and 0.42 for deuteranopes. 4. For colour normals the LWS cone fraction was 0.56, in agreement with psychophysical literature. The LWS cone fraction for protanopes was -0.04, and for deuteranopes 0.96, consistent with their Rayleigh matches. PMID- 8730605 TI - Nutrition survey of the Hungarian population in a randomized trial between 1992 1994. AB - OBJECTIVE: Collection of data relating to nutrition, nutrient supply and nutritional status, and to certain risk factors of diet-related diseases in a group of Hungarian population. DESIGN: Collection of data on diets three times 24h dietary recall, anthropometric measurements to obtain information about the nutritional status, laboratory tests measurements of biochemical markers. SUBJECT: Examination of 2559 volunteer adult persons, selected in a quasi random way in Budapest and seven other counties. RESULTS: The diets of the persons examined contained more than necessary protein 14.6 en%, considerably more fat 38.0 en%, mainly fat of animal origin with low PUFA and including high cholesterol, little total carbohydrate 45.1 en%, in that much added sugar 15.4 en%. Vitamin supply was generally satisfactory, borderline deficiency may be observed in the case of tocopherol equivalent and in some age-groups, first of all in women, in the case of thiamine, riboflavin and pyridoxine. Out of the macro- and microelements, sodium intake is three to four times the acceptable level, potassium and calcium intakes in both genders, iron and zinc intakes in women show deficiency. The mean value for relative body mass, the distribution of BMI values prove a high incidence of overweight 34.3% and obesity 21.1%. Means for biochemical markers appear to indicate that nutrient supply was in the normal range of laboratory tests. The averages of total cholesterol and LDL-cholesterol 5.6 and 3.6 mmol/L, respectively unequivocally suggested the risk of cardiovascular diseases. CONCLUSIONS: The nutrition of the Hungarian population presents several risk factors of diet-related diseases: P/S ratio of 0.27-0.29, Na/K ratio of 2.6 g/g, intake of some micronutrients (vitamins, macro- and microelements) was low in certain groups of investigated persons. PMID- 8730606 TI - Effects of moderate consumption of red wine on platelet aggregation and haemostatic variables in healthy volunteers. AB - OBJECTIVE: To evaluate the effect of moderate consumption of red wine on platelet aggregation and haemostatic variables, discriminating the effect of alcohol from that of non-alcoholic components. DESIGN: A randomised crossover study. SETTING: The Department of Food Science and Technology, University of Milan. SUBJECTS: Eleven healthy male volunteers who were moderate drinkers. INTERVENTIONS: For three periods of four weeks, subjects drank three different beverages [320 ml of red wine (providing 30 g/day of alcohol), 30 g/day of alcohol diluted in 320 ml of clear fruit juice or 320 ml of dealcoholised red wine] during the two main meals. Each treatment was preceded by a period of four weeks of complete withdrawal from any alcoholic beverage. At the end of each period platelet aggregation after collagen and ADP stimulus, and levels of fibrinogen, plasminogen, tissue-type plasminogen activator (t-PA) antigen and von Willebrand factor (vWF) were determined. RESULTS: Consumption for a period of four weeks of 30 g/day of alcohol either from red wine or alcohol resulted in similar decreases of collagen-induced platelet aggregation and fibrinogen levels. ADP-induced platelet aggregation, t-PA antigen, vWF and plasminogen levels were not affected by any treatment. No differences were detected when we compared platelet function and the other haemostatic variables at the end of red wine and dealcoholised treatments with findings at the end of alcohol treatment and abstinence. CONCLUSIONS: The well known positive effect of moderate consumption of red wine on haemostasis seems due to alcohol and not to the non-alcoholic fraction present in red wine. PMID- 8730607 TI - Consumption of reduced-fat products: effects on parameters of anti-oxidative capacity. AB - OBJECTIVE: Dietary fat intake is higher than recommended in most western countries and is associated with the prevalence of cardiovascular diseases, obesity and cancer. The growing public concern about the adverse effects of a high fat intake has led to an increased availability of ?reduced-fat' products. Consumption of reduced-fat products might be a convenient way to reduce the energy intake. Besides the potential of reduced-fat products to reduce the energy intake, it might change dietary fat intake, both qualitatively and quantitatively, which in turn might also affect oxidative stress, i.e. the anti oxidant/pro-oxidant ratio. In this paper we present the effects of 6-month consumption of reduced-fat products on body weight, energy, macronutrient and vitamin E intake, concentration of the plasma lipid peroxidation product malondialdehyde (MDA), erythrocyte free radical scavenging enzymes activities (glutathione peroxidase activity (GSH-Px); superoxide dismutase (SOD) and catalase) as well as plasma fat-soluble anti-oxidative vitamin concentrations (beta-carotene, lycopene and alpha-tocopherol). DESIGN: A randomized semi controlled parallel comparison trial of six months, preceded by a one-month adaptation period. SETTING: The reduced-fat and full fat products were provided through a realistic shop in order to mimic a ?free-living' situation. SUBJECTS: Subjects were recruited from respondents on advertisements. Eighty healthy, normal weight (BMI: range 20-30 kg/m2) subjects, aged between 20 and 55y, were selected to participate in this study; 76 subjects completed the study. INTERVENTIONS: 6 months consumption of either reduced-fat or full-fat products. RESULTS: During the intervention period, consumption of reduced fat products resulted in significantly lower energy intake, in a lower percentage of energy derived from saturated fatty acids, monounsaturated fatty acids and polyunsaturated fatty acids as well as in lower vitamin E intake, when compared to the control group. Body weight, the ratio polyunsaturated fatty acids/saturated fatty acids, plasma MDA, vitamin concentrations (beta-carotene, lycopene and alpha-tocopherol), and erythrocyte free radical scavenging enzyme activities (SOD, GSH-Px and catalase) were not affected by the intervention. CONCLUSION: Neither the difference in energy intake, nor the change in dietary fat composition seems to affect the integrity of the antioxidant scavenging capacity, assessed by measuring plasma MDA and antioxidative vitamins (lycopene, beta-carotene and alpha-tocopherol) and erythrocyte free radical scavenging enzymes (SOD, GSH-Px and catalase). PMID- 8730608 TI - Validation of a short food frequency questionnaire for assessment of dietary calcium intake in women. AB - OBJECTIVE: To assess the ability of a short food frequency questionnaire to measure dietary calcium intake in women. DESIGN: Estimates of calcium intake from the food frequency questionnaire were compared with those from seven-day estimated records in 58 Caucasian women aged 25-49 years in Dunedin, New Zealand. SUBJECTS: Subjects were recruited through poster and newspaper advertising around the central business district of Dunedin, New Zealand. Of 69 subjects initially recruited, nine failed to complete diet records and two were excluded on the basis that their diet record was unlikely to represent habitual intake. RESULTS: Mean dietary calcium intakes were 808.1mg/day from the estimated record and 776.9mg/day from the questionnaire. The mean difference in intake by the two methods (31.1mg, SD = 234.0, 95% CI = -30.4-92.7, p = 0.3) did not differ significantly from zero. The questionnaire classified women consuming less than 800mg calcium per day with 78% specificity. 81% of subjects when classified by the records fell into the same or adjacent quartiles when classified by the questionnaire. Only two subjects were grossly misclassified. CONCLUSIONS: The short food frequency questionnaire could be used to assess mean dietary calcium intake in young to middle aged women. The specificity of the questionnaire is high in identifying women who consume < 800mg calcium/day, therefore it could be used to target interventions for those with intakes below this level. PMID- 8730609 TI - Alpha-linolenic acid in rapeseed oil partly compensates for the effect of fish restriction on plasma long chain n-3 fatty acids. AB - OBJECTIVE: To examine the ability of alpha-linolenic acid (ALA) in low erucic acid rapeseed oil (RO) to compensate for the effects of a restriction in fish intake on plasma fatty acid composition. DESIGN AND SUBJECTS: Two times 6 weeks' randomized dietary intervention was used with blind crossover design in 40 healthy unconfined women and men (age 20-46y). INTERVENTIONS: Subjects were assigned to two fish restricted diets, namely RO diet and Trisun-sunflower oil (TSO) diet, with similar proportions of saturated : monounsaturated : polyunsaturated fatty acids (11.5:17.5:8.5% of total energy, En%), but differing in their ALA content (2.2 and 0.3 En%) and n-6 : n-3-ratio (3 : 1 and 23 : 1, respectively). The fatty acid compositions of plasma triglycerides (TG), cholesterol esters (CE), and phospholipids (PL) were analyzed by gas chromatography. Dietary intake was evaluated based on 3- to 7-day food records. RESULTS: The proportion of TG and CE ALA decreased on the TSO diet (from 1.6% to 0.9% and from 0.9% to 0.4%, respectively, P < 0.001) and increased on the RO diet (from 1.7% to 3.4% and from 0.9% to 1.3%, respectively, P < 0.001) compared to the baseline level. The proportion of eicosapentaenoic acid (EPA) in all three plasma fractions decreased on the TSO diet but not on the RO diet. The proportions of docosa-hexaenoic acid (DHA) decreased on both experimental diets and there was no difference in CE DHA between the diets. PL docosa-pentaenoic acid (DPA) and PL DHA remained at a higher level on the RO diet compared to the TSO diet (P < 0.001 and P < 0.05, respectively). CONCLUSIONS: ALA is metabolized to EPA in humans to a significant extent. The degree to which rapeseed oil (ca 50g/day) affects the proportion of EPA resembled the effect of a weekly portion (50-100g) of fatty fish depending on the fat content of the fish. PMID- 8730610 TI - The determinants of plasma beta-carotene: interaction between smoking and other lifestyle factors. AB - OBJECTIVE: To assess the effect of cigarette smoking on the relationship between dietary intake and circulating levels of beta-carotene. DESIGN: A secondary analysis of data collected for the Diet and Nutritional Survey of British Adults by The Department of Health and The Ministry of Agriculture, Fisheries and Food; the study was a cross-sectional survey of a representative sample of the British population. SETTING: The study was conducted on a free living adult (16-64 years of age) population resident in Great Britain. SUBJECTS: 1483 subjects completed a seven day weighed inventory and provided a blood sample. RESULTS: Smokers had statistically significantly lower dietary intakes and circulating levels of beta carotene than non-smokers. At the same level of dietary intake, smokers were more likely to have lower circulating levels than non-smokers. For every 1000 micrograms change in dietary intake, there was a 0.01 mumol/l change in plasma beta-carotene in smokers and a 0.04 mumol/l change in non-smokers. The effect of smoking on circulating levels of beta-carotene persisted after adjusting for other factors. CONCLUSIONS: The results suggest that smoking places extra demands on the host; the higher risk of heart disease for smokers may be due to a combination of an increased inflammatory response, reduced host defences, and increased levels of circulating LDL. Failure to take account of the interaction between risk factors may distort the estimates of the relative importance of each factor when considered in isolation. PMID- 8730611 TI - Reported alcohol intake, diet and body mass index in male smokers. AB - OBJECTIVE: The study was carried out to determine associations of reported alcohol intake with diet and body mass index. Type and frequency of consumed alcohol were also considered. DESIGN: A cross-sectional study. SETTING: The baseline examination of the participants of the Alpha-Tocopherol, Beta-Carotene Cancer Prevention Study in 1985-1988. SUBJECTS: 27215 middle-aged Finnish male smokers. INTERVENTIONS: The diet was assessed by a self-administered food use questionnaire: Subject's habitual diet and alcohol intake over the previous 12 months were asked. Body mass index was used as the measure of adiposity. RESULTS: Energy intake from food was not related to alcohol intake. Although alcohol consumption was associated with food selection (eg berry and coffee consumption), this only slightly influenced daily nutrient intakes. Intake of spirits was more consistently related to higher body mass index than that of other alcoholic beverages. Daily alcohol intake had a much smaller association with body mass index than less frequent use, independently of the total consumption. CONCLUSIONS: The differences in nutrient intake between abstainers, light and moderate alcohol consumers were small although the consumption of many foods varied with alcohol consumption. Even if alcohol consumption is one noteworthy factor associated with weight, the energy from alcohol increases body weight less than expected; both the type and frequency of consumed alcohol may explain why energy from alcohol is utilized less efficiently than non-alcoholic energy. PMID- 8730612 TI - Major fruit and vegetable contributors to the main serum carotenoids in the Spanish diet. AB - OBJECTIVE: To identify the main sources of serum carotenoids (lutein, zeaxanthin, lycopene, beta-cryptoxanthin, alpha-carotene and beta-carotene) among the fruits and vegetables in the Spanish diet, to be used in the design of food questionnaires and to provide useful information for epidemiological and comparative studies. DESIGN: A Family Budget Survey from the National Institute of Statistics (1990-1991); an open questionnaire dealing with foods consumed over a one-week period; physical amounts consumed annually and during three-month periods. Calculation of the percentage contribution of each food on the basis of a standard portion as well as the carotenoid concentrations previously reported by our laboratory. SETTING: Nationwide (urban and rural). SUBJECTS: Twenty-one thousand, one hundred and fifty-five (21 155) randomly selected, private households throughout Spain (72279 subjects). RESULTS: Nine vegetables (potato, tomato, green bean, green and/or red pepper, carrot, artichoke, spinach, lettuce and beet) and five fruits (orange, tangerine, banana, peach and watermelon) account for more than 96% of the intake of the major six carotenoids in serum, both on a yearly and on a seasonal basis. Fruit and vegetables provide between 3.0mg (in autumn) and 4.3mg (in summer) of these carotenoids per day. CONCLUSIONS: In our population, a small number of fruits and vegetables can provide significant information on carotenoid intake (both annually and seasonally) to be used in the design of questionnaires for epidemiological studies, minimizing misclassification and errors. The marked seasonality of some products causes wide differences in the supply of, above all, beta-cryptoxanthin and lycopene. PMID- 8730613 TI - An experimental estimate of the maintenance energy requirement in children with cystic fibrosis. AB - OBJECTIVE: To determine experimentally the maintenance energy requirement MER of children with Cystic Fibrosis (CF). DESIGN: Paired energy balance studies were undertaken on eight children with CF during an infection exacerbation, each child was his own control. SETTING: Paired energy balance (EB) studies at home over three days. SUBJECTS: Children known (from clinic records) to be susceptible to frequent infections were contacted. Four eligible children volunteered and completed two studies. INTERVENTION: Energy intake was estimated by weighted dietary inventory during days 1-3 and 16-18 of convalescence and obligatory energy losses determined by analysis of synchronous faecal and urinary collections using bomb calorimetry and urea concentration respectively Resting energy expenditure (EE) was determined using the DeltatrakTM100 Metabolic Monitor Apparent energy balance (B) was calculated by difference between metabolizable energy intake ME and EE such that B = ME - EE. RESULTS: The linear relationship between B (y) and ME (x) is given by y = 0.85x - 206.6 (1) when B = 0, ME = MER with a value of 243.1kJ/kg/24h(95% CL 136.9 and 410.3kJ/kg/24h). CONCLUSION: When MER = 243.1 + 167.2 - 106.2kJ/kg/24h and ME = 93% of gross energy (GE), a GE intake of 261kJ/kg/24h(95% CL of 440kJ/kg/24h) meets the MER, and negative EB is avoided. Agreement with previous estimates of MER suggests no enhancement of MER in CF. PMID- 8730614 TI - Triglycerides, fatty acids, sterols, mono- and disaccharides and sugar alcohols in human milk and current types of infant formula milk. AB - OBJECTIVE: To investigate differences in the fatty acid composition, sterols, minor carbohydrates and sugar alcohols between human and formula milk. DESIGN: We analyzed the concentrations of triglycerides, sterols, di- and monosaccharides and sugar alcohols, as well as the fatty acid composition of 10 currently available types of formula milk for term babies. Results were compared with mature human milk from 99 exclusively breast-feeding Dutch women, who collected 24-hour samples in the second week (n = 99), sixth week (n = 99) and 3 months (n = 25) after delivery. Infant formula milk data were considered different if they fell outside the mean +/- 2s.d. range of corresponding human milk data. RESULTS: The triglyceride concentrations in human milk were lower than those of the formula milk, possibly due to an incomplete collection of fat-rich hindmilk. Formula milks tended towards a higher proportion of medium chain fatty acids and lower proportions of longer-chain polyunsaturated fatty acids. Formulas had cholesterol concentrations 3-35 times lower, and much higher phytosterol concentrations, compared with the human milk. In the formula milk types the glucose, sorbitol and myoinositol concentrations were generally lower, whereas the fucose and erythreitol concentrations were in the lower mean +/- 2s.d. human milk range. The galactose concentrations in the formulas were generally higher. CONCLUSIONS: Formula milk and human milk differ considerably in fatty acid composition and concentrations of cholesterol, phytosterols, monosaccharides and sugar alcohols. The biological consequences of these differences in composition are uncertain. PMID- 8730615 TI - A 15N-leucine-dilution method to measure endogenous contribution to luminal nitrogen in the human upper jejunum. AB - OBJECTIVE: This study was done to investigate whether an intraveinous infusion of 15N-leucine was accurate to differentiate between endogenous and exogenous nitrogen in the human jejunum after meal ingestion. SUBJECTS: Four healthy human volunteers equipped with an upper jejunal tube. INTERVENTIONS: The jejunal effluents were collected both under fasting conditions and after ingestion of 300g of yoghurt. The nitrogen, amino acid composition and 15N-leucine enrichment were determined in the digesta. RESULTS: During fasting, the jejunal flow rates (mmol/h) of both total nitrogen and amino acids were stable (6.9 +/- 2.7 and 1.88 +/- 0.79, respectively). After yoghurt ingestion, the flow rate of total nitrogen increased to 28.6 +/- 5.8mmol/h at 2h. The 15N-leucine enrichment in plasma reached a plateau at 4.3 mole % excess after one hour and did not vary significantly after meal ingestion. The 15N-leucine enrichment of the endogenous secretion (Ee) in the jejunum was fitted by the equation: Ee = 2.18[1 - 2.05 x exp( - 0.42 x t)]. After yoghurt ingestion, the enrichment in jejunal secretions decreased during the first 80 min. The endogenous nitrogen, calculated from the 15N-enrichment, significantly increased from 20 to 40min after meal ingestion compared to the basal value (P < 0.05). The estimation of the exogenous nitrogen and amino acid yield 300min after yoghurt ingestion indicated that 62 +/- 30% of the exogenous nitrogen and 75 +/- 12% of the amino acids were absorbed in the upper jejunum. CONCLUSIONS: The 15N-leucine-dilution method appears to be a convenient method to differentiate between the exogenous and endogenous contributions to nitrogen fluxes in the intestinal digesta of humans. It can be used in association with dietary protein labelling or in substitution when no labelled dietary proteins are available to compare the digestion as well as the absorption of meals at different levels of the intestine. PMID- 8730616 TI - Effects of Bifidobacterium sp fermented milk ingested with or without inulin on colonic bifidobacteria and enzymatic activities in healthy humans. AB - OBJECTIVE: To assess in healthy humans the effects of prolonged ingestion of Bifidobacterium sp fermented milk (BFM) with or without inulin on fecal bifidobacteria and some bacterial enzymatic activities. DESIGN: Twelve volunteers randomly divided into two groups were studied for three consecutive periods. During the ingestion period, they received BFM in association with ether 18g/d inulin or placebo in three oral doses for 12 days. Stools were regularly collected for bacteriological analysis. SETTING: Clinical Nutrition Unit, Hopital Saint-Lazare, Paris. RESULTS: The administration of BFM with placebo led to an increase in total bifidobacteria (indigenous and exogenous) (P < 0.01) and exogenous bifidobacteria (P < 0.01) and a decrease in beta-glucuronidase activity (P < 0.01). Simultaneous administration of BFM and inulin led to an increase in total bifidobacteria (P < 0.01) and exogenous bifidobacteria (P < 0.01), but had no effect on beta-glucuronidase activity. No differences were found for fecal concentrations reached by exogenous and indigenous bifidobacteria between the two groups. Administrated alone or with inulin, BFM did not change fecal total anaerobe counts, pH, nitrate reductase, nitroreductase and azoreductase activities. CONCLUSIONS: Administration of BFM substantially increases the proportion of bifidobacteria in the colonic flora, but the concurrent administration of inulin does not enhance this effect. PMID- 8730617 TI - Effects of oat gum supplementation on blood lipid fractions. PMID- 8730618 TI - Perinatal morbidity and mortality in offspring of diabetic mothers in Qatif, Saudi Arabia. AB - Diabetic mothers and their offspring were prospectively studied. Perinatal and neonatal morbidity and mortality data were analysed. Out of 11,677 deliveries in the hospital, 133 (1.14%) were delivered by diabetic mothers. They were made up of six (4.5%) stillbirths and 127 (95.5%) live births. Seventy-seven (57.9%), 55 (41.3%) and 1 (0.8%) were large, appropriate and small for gestational age, respectively. Hypoglycemia occurred in 49 (38.6%) of the 127 infants. Other associated problems were bacterial infections, congenital anomalies, birth trauma, preterm delivery, respiratory distress, polycythaemia and anaemia. Only 52 (39.1%) of the mothers received appropriate treatment for their diabetes during pregnancy. Poor maternal diabetic control resulted in high perinatal morbidity and mortality in the offspring. In order to improve the outcome in offspring of diabetic mothers in Qatif and probably Saudi Arabia as a whole, health education and improved care of the diabetic mothers during pregnancy urgently needed. This may be true of other developing countries where data on diabetes in pregnancy are scarce. PMID- 8730619 TI - Bone density studies in pregnant women receiving heparin. AB - Lumbar spine bone density falls during pregnancy in both women who are treated with heparin and those who are not. Breast feeding delays recovery to baseline values until suckling stops. PMID- 8730620 TI - Uterine rupture during second trimester termination of pregnancy using mifepristone and a prostaglandin. PMID- 8730621 TI - Hyperreactio luteinalis masquerading as an ovarian neoplasm in a triplet pregnancy. AB - Hyperreactio luteinalis is a non-neoplastic tumor-like ovarian lesion associated with pregnancy. Most patients are asymptomatic, with the ovarian enlargement being incidentally discovered at the time of cesarean section. It can simulate a neoplasm on clinical, gross and sometimes microscopic examination. We report a case of hyperreactio luteinalis in a patient, who was diagnosed as having polycystic ovary disease before conceiving a triplet pregnancy after three treatment cycles of human menopausal gonadotropin-human chorionic gonadotropin therapy, and discuss its pathogenesis. PMID- 8730622 TI - Prenatal exposure to diethylstilbestrol and the mother-daughter relationship. AB - The psychological consequences resulting from the exposure to diethylstilbestrol (DES), a non-steroidal oestrogen, on the mother-daughter relationship are studied using semi-directive interviews with 43 daughters and 7 mothers treated with DES during their pregnancies. These women referred to gynaecological consultation for DES-related problems. The daughters, exposed to DES during their foetal life, learned about DES after a pregnancy mishap (35% of the cases), or by accident (65% of the cases). All of them were shocked when the existence of DES and its side effects were revealed to them. Consequences on the mother-daughter relationship were absent in 60% of the cases, favourable in 20%, and negative in 20%. Five percent of the women showed hostility towards the medical practice, but 65% were not suspicious of the drugs administered to them during their pregnancies. For 64% of them, administration of DES to their mother had been kept secret. In 7 out of 50 cases, parents alone came for medical assistance in order to manage the secret. Exposure to DES may reveal pre-existing difficulties not only between the mother and the daughter, but sometimes beyond from generation to generation. PMID- 8730623 TI - Hysteroscopic management of uterine synechiae: a series of 102 observations. AB - This retrospective study concerns 102 cases of intra-uterine adhesions treated by exclusive hysteroscopic approach. Fifty-two patients (51%) presented with reproductive problems, 44 patients (43%) with menstrual problems and 6 were asymptomatic. Seventy-eight patients (76.5%) had an antecedent endometrial trauma on a gravid uterus. One hundred and forty-eight operative hysteroscopies were performed to treat 102 patients; 70 patients were treated in one endoscopic session (68.6%), 23 in two sessions (22.6%), 4 in three sessions (3.9%) and 5 in four sessions (4.9%). We report no infectious, hemorragic or metabolic complication but 6 perforations (5.8%). The mean follow-up is 24.4 months (from 6 to 46 months). Ten patients did not keep contact during follow-up. A good anatomical result was obtained in 88 patients (86.2%) after one or two hysteroscopic sessions. A good result on menstrual problems was obtained in 75% of the cases, especially in amenorrhea (90.5% success rate). The reproductive outcome is more disappointing. We obtained 34 pregnancies in 28 patients (10 abortions, 24 live born children). Twenty-two patients achieved no pregnancy (44%), but 13 of these patients presented with additional infertility factors. PMID- 8730624 TI - Cervical intra-epithelial neoplasia in women infected with human immunodeficiency virus. AB - OBJECTIVE: To investigate the relationship between human immunodeficiency virus (HIV) and cervical intra-epithelial neoplasia (CIN). DESIGN: A prospective study to determine the prevalence of cervical intra-epithelial neoplasia in 111 HIV positive women. METHODS: In total, 111 HIV + women were enrolled and underwent cervical biopsy and CD4 T-lymphocyte determination. Of the 111 women, 26 (23.4%) had CIN and another 26 (23.4%) had histologic evidence of cervicitis. RESULTS: No significant difference was found between transmission group, CDC stage of disease, CD4 T-lymphocyte count and pregnancy. There was a significant association with concomitant human papillomavirus (HPV) infection (P < 0.001). CONCLUSION: Public health measures are needed to provide Papanicolaou smear screening and appropriate clinical follow-up and treatment of women infected with the human immunodeficiency virus. PMID- 8730625 TI - Adjuvant progestagen therapy for the treatment of endometrial cancer: review and meta-analyses of published randomised controlled trials. AB - OBJECTIVE: To assess the efficacy of adjuvant progestagen therapy for the treatment of endometrial cancer. DESIGN: Computerised literature review by MEDLINE backed up by a manual search of 16 journals published before January 1995, to identify relevant published randomised controlled trials. Trials were classified according to methodological quality criteria. Odds ratios and 95% confidence intervals (CIs) were calculated where data allowed. SETTING: Institute of Epidemiology and Health Service Research, University of Leeds. SUBJECTS: Patients presenting with endometrial cancer. INTERVENTION: Adjuvant progestagen therapy. OUTCOMES: Death (overall, cancer related and intercurrent) and relapse. RESULTS: Six randomised controlled trials comparing adjuvant progestagen with no adjuvant therapy in the treatment of endometrial cancer were identified in the literature prior to January, 1995. Three trials had an acceptable method of randomisation. Meta-analysis demonstrated that overall survival was not improved by adjuvant progestagen therapy and may even have been adversely affected. Furthermore, this treatment was not associated with any reduction in relapse or death from endometrial cancer. CONCLUSION: Current evidence does not support use of adjuvant progestagen therapy in the primary treatment of endometrial cancer. PMID- 8730626 TI - Fertility and pregnancy outcome after myomectomy in sterility patients. AB - OBJECTIVES: To examine total pregnancy rate, pregnancy rate in relation to pretreatment with GnRH-analogues, the frequency of myoma recurrencies and the influence of size, number and localization of removed myomata on pregnancy rate and outcome in infertility patients after myomectomy. STUDY DESIGN: A comparative, retrospective non-randomized clinical study involving 67 patients with desire for children and no other recognizable infertility factor. Myomectomy had been performed between 1985 and 1993. Most patients had been operated by laparotomy using microsurgical instruments and techniques. Thirty-three patients had been treated with a GnRH agonist for usually 3 months, and in 34 patients the operation was performed without pretreatment. Patients were followed up to June, 1994. All patients were mailed a questionnaire and invited to an ultrasound examination. RESULTS: Thirty-nine of the 67 patients (58.2%) became pregnant, and a total of 51 pregnancies were observed. Of the women who actually conceived, 61.5% did so within the first year. There was no significant difference in pregnancy rates between patients who had been pretreated with GnRH agonists and those who had not. However, 1 year after the operation the group of GnRH-treated women was significantly overrepresented among those already pregnant (P = 0.02). Sonografical examination revealed in 31 out of the 67 patients (46.3%) recurrent myomata > 1 cm in diameter. There was no statistically significant difference in the pregnancy rates between patients with and without recurrencies. However, there was a significant tendency toward a loss or short duration of the pregnancy due to spontaneous abortion and premature delivery in patients with recurrent or persistent myomata (P < 0.01). Pregnancy rate was significantly lower in patients with more than five myomata removed (P < 0.001). In the group with a larger myoma volume the pregnancy rate was significantly higher than in the group with the smaller one (P < 0.01), possibly indicating that the size on removal of myomata is an important factor for infertility patients. Concerning the localization of the removed myomata, no statistically significant difference was found in the pregnancy rates between various localizations. Of the 51 pregnancies, 31 (60.8%) led to a delivery, vaginal in 13 cases (41.9%) and 18 times by Caesarean section (58.1%). Of the pregnancies that were lost, 39.2% were due to spontaneous abortion or ectopic pregnancy. CONCLUSIONS: Our study supports reports on the benefits of myomectomy, performed with the appropriate technique, in patients with otherwise unknown cause of infertility. It shows, additionally, that characteristics of myomata, such as number and size, may influence postmyomectomy pregnancy rates. PMID- 8730627 TI - Experience with laparoscopic assistance in vaginal hysterectomy. AB - OBJECTIVE: To determine safety and usefulness of laparoscopy intended to convert an abdominal into a vaginal hysterectomy. STUDY DESIGN: Forty-seven consecutive hysterectomies are retrospectively analyzed in which laparoscopy was used to accomplish a vaginal instead of an abdominal procedure which would have been performed without endoscopic surgery. RESULTS: In four patients the operation had to be converted to laparotomy due to difficult access (n = 3) or heavy bleeding (n = 1). Operation time declined during the study period, thus highlighting the importance of experience with laparoscopic technics. Major complications such as injuries of adjacent organs or delayed haemorrhage requiring a second intervention were not encountered. CONCLUSIONS: Laparoscopy in hysterectomy seems to be a valuable method to avoid laparotomy. In the future, guidelines to determine the route of surgery are to be established which include those conditions in which laparoscopy in addition to vaginal hysterectomy implies a benefit for the patient. PMID- 8730628 TI - A combined approach for the early detection of ovarian cancer in asymptomatic women. AB - The purpose of this study was to investigate the effectiveness of the combined procedure including pelvic examination and serum CA-125 determination, as a screening method for the early detection of ovarian cancer. In 2000 women over 45 years old, without any evidence of adnexal disease, pelvic examination and serum CA-125 determination were performed. When either the findings of the pelvic examination were ambiguous or positive, or the serum CA-125 level was > 35 U/ml, further investigation including ultrasonography and laparoscopy or laparotomy was done. Among 174 women with clinical findings of adnexal disease there were 15 (8.62%) who had serum CA-125 > 35 U/ml. Among 18 women with elevated serum CA-125 (> 35 U/ml) there were 15 women (83.33%) who had clinical findings of adnexal disease. In 15 women further investigation was suggestive of adnexal disease and surgical exploration revealed three cases of malignancy. One case of serious ovarian cystadenocarcinoma stage Ia, one case of borderline ovarian tumour and one case of metastatic carcinoma from the right kidney. The other 12 women had benign adnexal masses or pelvic endometriosis. In our study this combined approach had sensitivity 100%, specificity 99.70% and P.P.V. 22%. According to these data, the procedure could be a potential component of a strategy aimed to the early detection of ovarian cancer, regardless of the financial cost. PMID- 8730630 TI - Regulation of transferrin receptor synthesis by human cytotrophoblast cells in culture. AB - The aim of this study was to examine the capacity of the syncytiotrophoblast to regulate transferrin receptor (TfR) synthesis in response to modulations in maternal iron supply. The model used was the primary trophoblast cell culture. Trophoblast cells isolated from term human placentas were cultured in iron-poor (Medium 199), iron-depleted (desferrioxamine (DFO)) and iron-supplemented (diferric transferrin (hTf-2Fe), ferric ammonium citrate (FAC) medium. TfR synthesis was reduced in response to hTf-2Fe supplementation. FAC did not modulate TfR synthesis. Iron deprivation by DFO resulted in clear stimulation of TfR synthesis. These results show that the differentiating trophoblast cells respond to pertubations in the (transferrin-mediated) iron supply by adjustments in the rate of TfR synthesis. Taking syncytiotrophoblast in culture as model for the maternal/fetal interface in vivo, our results would suggest that the placenta is able to make short term adjustments of the capacity for iron uptake. PMID- 8730629 TI - Corticosteroid treatment does not improve the results of intrauterine insemination in male subfertility caused by antisperm antibodies. AB - AIM: To determine whether corticosteroid immunosuppression (CS) administered to the male partner together with intrauterine insemination (IUI) is preferable compared to IUI alone in treating male autoimmune subfertility. MATERIALS AND METHODS: Thirty-six couples with proven male immunological subfertility were randomly assigned to begin CS + IUI (n = 18) or IUI (n = 18) treatment and progressed alternatively to receiving each treatment modality for three cycles unless pregnancy occurred. Each couple served as their own control. The administered corticosteroid was soluble prednisolone. RESULTS: Five pregnancies were achieved with 77 cycles of CS + IUI and seven pregnancies with IUI alone. The pregnancy rates per cycle were 6.5% with CS + IUI and 9.21% with IUI, while the pregnancy rates per couple were 16.13% with CS + IUI and 21.2% with IUI alone. These rates do not differ to a statistically significant degree (P > 10%). CONCLUSIONS: The addition of corticosteroid immunosuppression does not seem to significantly enhance the pregnancy rate in couples with male autoimmune subfertility treated with IUI as compared to the treatment with IUI alone. Considering the possible side-effects of corticosteroid intake, we conclude that IUI alone might be preferable to the combination of IUI with corticosteroid administration. PMID- 8730631 TI - Management of leiomyoma causing myocardial infarction. AB - Although most myocardial infarctions are caused by either obstruction or spasm of the coronary arteries, severe anemia can lower oxygen carrying capacity and damage the myocardium. We report a case of leiomyoma-induced menorrhagia of such severity that myocardial infarction ensued. Hysterectomy was indicated to avoid further anemia. While delaying elective surgery until the heart has had time to recover from infarction is the usual practice, the fact that myoma-induced menorrhagia contributed to ischemia in this patient prompted us to perform hysterectomy shortly after infarction. PMID- 8730632 TI - A case of life-threatening pulmonary edema associated with amnioinfusion during labor. PMID- 8730633 TI - Septate uterus with cervical duplication and longitudinal vaginal septum: a report of three new cases. AB - The first case of a rare mullerian anomaly characterized by the presence of a complete uterine septum with duplication of the cervix and a longitudinal vaginal septum has been reported very recently. We present here three new cases of such an anomaly in an attempt to alert gynaecologists to the possible occurrence of such a malformation. The cases challenge the classical views of unidirectional (caudad to cranial) mullerian development and support the alternative embryologic hypothesis of Muller et al. according to which fusion and resorption begins at the isthmus and proceeds simultaneously in both the cranial and caudal directions. PMID- 8730634 TI - An acardiac acephalic monster following in-utero anti-epileptic drug exposure. AB - Acardia, the absence of the heart, is one of the rarest medical anomalies. The exact mechanism which causes this anomaly is still unknown. The authors report the acardiac acephalic fetus of an epileptic mother who was on primidone therapy. The mother who received no antenatal care stopped taking primidone (her sole medication) in the third month of pregnancy with the fear of delivering a malformed baby and had three convulsions until delivery. This is the first reported case of acardia associated with anti-epileptic medication. The cause of the anomaly in this patient may be an unknown genetic defect, the maternal epileptic disorder, the convulsions, the anti-epileptic medication, or a combination of these factors. PMID- 8730635 TI - Mixed mesodermal sarcoma of the ovary in a young patient. AB - Mixed mesodermal tumors (MMT) of the ovary are rare and have a poor prognosis. This ovarian malignancy usually occurs in postmenopausal women. We report an unusual ovarian MMT in a young woman given treatment similar to one used for ovarian germ cell malignancies. We believe this is the youngest patient reported with an homologous MMT of the ovary. PMID- 8730636 TI - Unmeasurable relaxin concentrations in repeated pregnancies after embryo transfer. AB - Unmeasurable serum relaxin concentrations were found in repeated serum samples taken during two subsequent pregnancies in a woman pregnant after in vitro fertilization in artificial cycles. Despite the absence of relaxin both pregnancies were carried out normally until term except for symptoms of pelvic girdle relaxation. Labour induction was attempted at term as placental insufficiency was suspected. However, the cervical score remained unfavourable and Cesarian section was performed. In the second pregnancy elective Cesarian section was performed due to mild preeclampsia. This case indicates that relaxin may not be important for either suppression of myometrial activity or pelvic girdle relaxation during human pregnancy. PMID- 8730638 TI - Somatization--beyond symptom count. PMID- 8730637 TI - Editorial: Psychosomatic research. 1956. PMID- 8730639 TI - The "railway spine" diagnosis and Victorian responses to PTSD. PMID- 8730640 TI - Screening for eating disorders in diabetes mellitus. AB - Eating disorders in diabetes mellitus may be associated with serious diabetic complications. This study examines the prevalence of complications and the usefulness of the Eating Attitudes Test (EAT) in screening for eating disorders in a group of insulin-dependent diabetic women. Coping strategies for dealing with diabetes are also investigated. Increased rates of diabetic complications and insulin manipulation were confirmed among subjects with eating disorders. The EAT alone had a poor predictive value for identifying eating disorders, but the presence of raised EAT score or insulin manipulation proved effective in identifying almost all cases of eating disorder. Subjects with high EAT scores showed coping styles characterised by acceptance-resignation. PMID- 8730641 TI - Dimensions of hostility and myocardial infarction in adult males. AB - The present case-control study investigated the association between dimensions of hostility and myocardial infarction (MI) in adult males. Hostility was measured with the Buss-Durkee Hostility Inventory (BDHI), which assesses two distinct dimensions of hostility, namely experiential and expressive hostility. Cases were 81 males who were admitted in hospital because of a first MI. The reference group consisted of 168 age-matched, healthy male neighbourhood controls. Analyses revealed that especially experiential hostility (comprised of the subscales Resentment and Suspicion) was significantly associated with MI. These results are in contrast with the findings of previous studies, which showed expressive hostility to be positively related to coronary heart disease (CHD). Explanations for these contradictory findings are discussed. PMID- 8730642 TI - General and illness-specific adjustment to cancer: relationship to marital status and marital quality. AB - This study examined the relationship between psychological adjustment of adults with cancer and two marital variables: married versus unmarried and high versus low marital quality. Eighty-six married and 46 unmarried adults with cancer completed general and illness-specific measures of psychological adjustment. In general, men and subjects with low marital quality reported more depression and anxiety, a less positive health care orientation, and more illness-induced family difficulties than did women and subjects with high marital quality; unmarried subjects reported more dysphoric thoughts and feelings related to the their illness than did married subjects; and unmarried men reported more disruption in work activities and extended family relationships than did unmarried females or married subjects. Moreover, a greater percentage of unmarried subjects and subjects with low marital quality reported clinically elevated levels of symptomatology compared to those with high marital quality. The clinical implications of these findings are discussed. PMID- 8730643 TI - Spouse-pair risk factors and cardiovascular reactivity. AB - Epidemiological research has identified increased risk for coronary heart disease in Type A men married to well-educated women. The present study examined mechanisms that may explain the increased risk associated with this specific spouse-pair combination. Cardiovascular and self-report responses to an individual, standardized laboratory stress task and a dyadic, interactive affect provocation task were assessed in Type A or B men married to women of either low ( < or = 13 years) or high ( > 13 years) educational levels. Type A men with highly educated spouses (i.e., the highest risk group) also reported the greatest anger-out tendency, high trait anger, and low anger control scores. These men further exhibited elevated diastolic blood pressure at baseline and greater diastolic reactivity specific to the dyadic task than did the men in the lower risk groups. These findings, based on a laboratory study of spousal interactions, support the epidemiological high-risk designation of the Type A man with a highly educated spouse and confirm earlier conceptions that Type A research benefits more from a social interaction approach rather than from an individual trait perspective. PMID- 8730644 TI - Is aspirin, as used for antithrombosis, an emotion-modulating agent? AB - Antiplatelet substances, generally aspirin, have become widely used for secondary prevention of ischemic heart disease. Used in relatively small doses, it is generally assumed that aspirin has no psychoactive effect. The present study took advantage of a sample of 174 males undergoing coronary angiography to see if regular aspirin use as prophylactic therapy for ischemic heart disease was associated with one or more of a number of measures of emotional distress. Aspirin use was found to be associated with less depression and anxiety or worry, as reported by the patient and as perceived by a significant other. Despite a significant association of aspirin use with the presence of documented coronary artery disease, the association of aspirin use and diminished distress could not be accounted for by the previously observed high prevalence of depressed/anxious individuals among patients with negative or nominal results on angiography, or by a number of other demographic or clinical variables such as age and socioeconomic status. Although only correlational in nature, present results raise the question of whether aspirin may have a beneficial mood-modulating effect. PMID- 8730645 TI - Depressive symptoms in overweight and obese older adults: a test of the "jolly fat" hypothesis. AB - The association between body weight and depressive symptoms in older adults was examined in a population-based study of 2,245 noninstitutionalized men and women aged 50 to 89 years living in Rancho Bernardo, California, U.S.A. The prevalence of Beck Depression Inventory scores > or = 13 was inversely associated with body weight in men, but not in women. Overweight and obese 50- to 69-yr-old women were more depressed than women with a body mass index below 25 kg/m2, but the difference was only marginally significant (p = 0.09). When age, health status and medication use were controlled, the odds of being depressed were 0.34 (p = 0.004) in overweight men and 0.28 (p = 0.09) in obese men, compared to men with a body mass index below 25 kg/m2. In this cohort, depression in men was inversely associated with body weight, supporting the "Jolly Fat" hypothesis. The likelihood that more stigma is attached to excessive weight in women than men may account for the lack of an inverse association between weight and depression in women. PMID- 8730646 TI - MMPI profiles of patients with chronic fatigue syndrome. AB - Fifty-three patients with chronic fatigue syndrome (CFS) and 43 healthy nonpatient controls completed the Minnesota Multiphasic Personality Inventory (MMPI). All subjects varied in their degree of seropositivity to active Epstein Barr virus (EBV) as measured by their anti-early antigen titers. EBV titers were higher among CFS patients and were associated with being more symptomatic. Differences in patient status were associated with statistically significant elevations on 8 of 9 clinical scales, 4 of which also showed clinically significant elevations (T scores > or = 70): scales 1, 2, 3, and 8. These results are discussed in terms of their implications for intervention strategies associated with MMPI-based CFS subtypes. PMID- 8730647 TI - Baseline correlates of the response to the treatment of chronic localized myofascial pain syndrome by injection of local anaesthetic. AB - Myofascial pain syndromes (MPS) occur commonly in the community. Treatment by injection of local anaesthetic has met with variable success. We studied 50 out patients with chronic localized MPS in order to find baseline correlates related to response to treatment. Data collected included sociodemographic and clinical histories as well as psychometric measures of self-efficacy, health locus of control and illness behaviour. Pretreatment pain intensity and scores on the Denial scale of the Illness Behavior Questionnaire (IBQ) were found to be associated with an immediate improvement in pain, while scores on the Affective Inhibition scale of the IBQ were associated with pain relief lasting up to 24 hours. The immediate response was not predictive of the duration of the effect of the treatment. This study shows that the severity of the pain and its role as part of a psychological coping strategy are related to the response to local treatment of MPS. PMID- 8730648 TI - The Patient Requests Form: a way of measuring what patients want from their general practitioner. AB - A consumer-oriented approach to the delivery of health care and an understanding of the processes that influence clinical management decisions require the measurement of what patients seek when they consult their doctor. The present study completed the development of an instrument (the Patient Requests Form) that can quantify the intentions of patients attending their general practitioner. The Patient Requests Form was completed by 410 patients attending two general practices: one in an inner city area, the other in a small town. Principal components analysis revealed that the responses from each sample yielded identical components that described three distinct types of request: (i) for explanation and reassurance, (ii) for emotional support, and (iii) for investigation and treatment. Scales constructed to measure each type of request have high internal consistency while being sufficiently brief to be acceptable to general practice patients. The Patient Requests Form is a novel, convenient method to quantify the intentions of patients when they consult a general practitioner. It permits research into neglected aspects of consultation behaviour, including the factors that influence patients' intentions to seek different kinds of help and GPs' perceptions of these intentions. PMID- 8730649 TI - A validation study of the Whitely Index, the Illness Attitude Scales, and the Somatosensory Amplification Scale in general medical and general practice patients. AB - The aim of this study was to assess the reliability and validity of the Whitely Index (WI), the Illness Attitude Scales (IAS), and the Somatosensory Amplification Scale (SAS). The study population consisted of 130 general medical outpatients, 113 general practice patients, and 204 subjects from the general population. The factorial structure of the IAS appeared to consist of two subscales, namely Health Anxiety and Illness Behaviour. The internal consistency and stability of the three questionnaires were satisfactory, and their scores were highly intercorrelated. Scores on the WI and Health Anxiety subscale of the IAS declined significantly from general medical outpatients, through general practice patients to subjects from the general population. This might imply that medical care utilisation is related to hypochondriasis. A prospective study is needed to determine whether health anxiety contributes to the decision to seek medical care or the consultation of a general practitioner or consultant gives rise to worry about possible illness. PMID- 8730650 TI - The role of oxytocin release in the paraventricular nucleus in the control of maternal behaviour in the sheep. AB - Oxytocin (OT) release within the brain is thought to play a major role in inducing maternal behaviour in a number of mammalian species but little is known about the sites of release which are important in this respect. We have investigated whether the paraventricular nucleus of the hypothalamus (PVN) is a site of OT action on maternal behaviour in the sheep. In vivo microdialysis and retrodialysis was used to determine whether OT is released in the region of the PVN during the post-partum induction of maternal behaviour and if its release at this site can stimulate maternal behaviour in non-pregnant animals. In vivo sampling showed that OT concentrations increased significantly in the region of PVN at birth. When OT was retrodialysed bilaterally into the PVN (1 or 10 microM) of multiparous ewes treated with progesterone and oestradiol to stimulate lactation, maternal behaviour was induced in a significant number of animals (1 microM, 6/8 and 10 microM, 5/8) compared with controls (0/8 ewes). Similar infusions of the ring structure of OT, tocinoic acid (TOC-10 microM), also induced maternal behaviour in a significant proportion of animals (5/6 ewes) as did intracerebroventricular (ICV) OT (6/8 ewes) and artificial stimulation of the vagina and cervix (VCS, 8/9 ewes). On the other hand, vasopressin (AVP) 1 microM did not induce maternal behaviour in any ewes and a 10 microM dose only induced it in 2/8 animals. The neurochemical changes accompanying the above treatments were also investigated. Noradrenaline concentrations increased in the PVN after the retrodialysis administration of OT 1 microM and 10 microM, TOC 10 microM and AVP 1 microM, OT ICV and VCS. Dopamine concentrations were also increased by OT 10 microM, TOC 10 microM, AVP 1microM and OT ICV. Aspartate and glutamate concentrations were significantly reduced by retrodialysis infusions of OT 1 microM and AVP 1 and 10 microM but not by any other treatment. Finally, the retrodialysis infusion of OT and TOC, as well as ICV OT, significantly increased plasma OT release whereas AVP infusions did not. These results provide evidence that OT is released in the PVN during parturition and is important for the induction of maternal behaviour. It seems probable that OT release at this site has a positive feedback effect on both parvocellular and magnocellular OT neurons to facilitate co-ordinated OT release both in central OT terminal regions (to facilitate maternal behaviour) and peripherally into the blood (to facilitate uterine contractions/milk let down). The potential functional roles for the actions of OT on monoamine and amino acid transmitter release in the PVN are discussed. PMID- 8730651 TI - Inhibition of steroid-induced galanin mRNA expression in GnRH neurons by specific NMDA-receptor blockade. AB - Galanin mRNA levels in GnRH neurons increase in association with a steroid induced LH surge in female rats. Both the steroid-induced LH surge and the concomitant increase of galanin mRNA in GnRH neurons are blocked by non-specific inhibition of central nervous system activity imposed by pentobarbital and specific central alpha-adrenergic receptor blockade. Based on these observations, we hypothesized that galanin gene expression in GnRH neurons is induced whenever GnRH neurons become activated to generate an LH surge. If this were the case, then any neurotransmitter receptor blocking agent that inhibits the LH surge by central mechanisms would likewise block the associated increase in galanin mRNA in GnRH neurons. We tested this hypothesis by examining the effects of an N methyl-D-aspartate (NMDA) receptor antagonist on the steroid-induced LH surge and on levels of galanin mRNA in GnRH neurons. Three groups of ovariectomized rats were used: Group 1 -treated with estradiol and progesterone (E/P) and sacrificed at the peak of the LH surge; Group 2-treated the same as Group 1 except that dizocilpine (MK801, an NMDA receptor antagonist) was used to block the LH surge; and Group 3-treated the same as Group 1 except they received vehicle instead of E/P. Double-and single-label in situ hybridization followed by computerized image analysis were used to measure levels of galanin mRNA and GnRH mRNA in GnRH neurons [as grains/cell (g/c)]. E/P treatment induced a 3-fold increase in LH levels and a 5-fold increase in the galanin mRNA signal content of GnRH neurons. Treatment with MK801 completely prevented the LH surge in all animals and also blocked the steroid-induced increase in galanin mRNA in GnRH neurons. As assessed by 2 independent GnRH single-labeled assays, neither GnRH message content nor the number of identifiable GnRH neurons differed among the experimental groups. We conclude that the increase in galanin mRNA levels in GnRH neurons is tightly coupled to the occurrence of a steroid-evoked LH surge, and we infer that induction of galanin gene expression in GnRH neurons is induced as a consequence of synaptic activation of GnRH neurons. PMID- 8730652 TI - Induction of galanin mRNA in GnRH neurons by estradiol and its facilitation by progesterone. AB - On the day of proestrus in the rat, rising plasma levels of estradiol (E) act in concert with progesterone (P) to trigger a preovulatory release of gonadotropins. Cellular levels of galanin mRNA in GnRH neurons are increased in association with the proestrous surge of gonadotropin secretion; however, the relative contribution made by E and P to the induction of galanin mRNA expression in GnRH neurons is unknown. We investigated the role of E and P in the induction of galanin gene expression in GnRH neurons by examining the effects of different combinations of E (estradiol benzoate; 50 micrograms and P (5 mg)) on the LH surge and the concomitant induction of galanin mRNA in GnRH neurons. We sacrificed ovariectomized adult rats after 1 of 4 treatments: Group 1: vehicle control (n = 6); Group 2: P alone (n = 7) Group 3: E alone (n = 7); Group 4: combined E/P (n = 6); the animals were killed at 18.00 h at the time of the LH surge. The brains from these animals were processed by double-label in situ hybridization to allow measurement of galanin mRNA levels in GnRH neurons. GnRH neurons were identified with a digoxigenin-labeled cRNA probe for GnRH mRNA, and galanin mRNA was detected and measured simultaneously with an 35S-labeled cRNA probe coupled with computerized grain counting. Estimation of cellular levels of GnRH mRNA was accomplished with single-label in situ hybridization, an 35S labeled GnRH cRNA probe and computerized grain counting. We observed a 3-fold induction of galanin mRNA in the GnRH neurons of animals treated with E alone compared with those treated with the vehicle alone (vehicle: 13 +/- 2 vs E: 42 +/ 4 grains/cell (g/c); P < 0.01); LH levels in the E-treated animals were elevated, albeit moderately, with respect to the vehicle controls. Compared with vehicle-treated animals, those treated with the combination of E and P showed a 5 fold induction of galanin mRNA in GnRH neurons (68 +/- 9 g/c), which was significantly (P < 0.01) greater than that observed in the animals treated with E alone; in addition, the magnitude of the LH surge was much greater (P < 0.05) in the E/P-treated group compared with the E alone group. In contrast, compared to the vehicle controls, animals treated with P alone (15 +/- 2 g/c) showed no discernable effect on galanin mRNA levels; moreover, no LH surge occurred in the P alone group. Neither the number of identified GnRH cells nor their content of GnRH mRNA differed significantly among the experimental groups (GnRH mRNA signal: vehicle controls: 153 +/- 6 vs E: 159 +/- 6 vs E/P: 153 +/- vs P: 148 +/- 8 g/c). We conclude that while E is the primary ovarian signal inducing galanin mRNA expression in GnRH neurons and the LH surge itself, P plays a facilitatory role in both of these processes. PMID- 8730653 TI - Sex differences in cytosolic androgen receptors in gonadectomized male and female rats. AB - The purpose of this experiment was to determine if there are sex differences in cytosolic androgen receptors (AR) in individual brain nuclei. Bilateral 500 mu diameter samples from 300 mu thick frozen brain sections were micropunched from males and females 2-3 weeks following gonadectomy. Tissue samples were taken from 12 brain nuclei: lateral septum, bed nucleus of the stria terminalis, medial preoptic nucleus, anterior hypothalamic area, arcuate nucleus, corticomedial nucleus of the amygdala, lateral preoptic area, parietal cortex, medial nucleus of the amygdala, dorsomedial nucleus of the hypothalamus, ventromedial nucleus of the hypothalamus, and dorsal hippocampus. Cytosolic ARs were found in brain regions involved in the neuromodulation of androgen dependent responses. These data suggest that behavioral differences in male reproductive responses may be the sexually dimorphic distribution of androgen receptor containing neurons. PMID- 8730654 TI - Effect of vorozole, an aromatase enzyme inhibitor, on sexual behavior, aromatase activity and neural immunoreactivity. AB - Aromatase enzyme is essential for the expression of normal sexual behavior in many mammals and birds. Here we report that vorozole (R83842), a non-steroidal aromatase inhibitor, blocks sexual behavior in the female musk shrew. In addition, vorozole treatment lowers aromatase activity in male and female preoptic area, and reduces plasma estradiol concentrations in females. Our findings confirm and extend results demonstrated in other species, conducted with the active enantiomer (R83842), or the racemic mixture (R76713, racemic vorozole). We also report that vorozole treatment affects the immunocytochemical distribution of aromatase immunoreactivity (AROM-ir) in musk shrew brain. The histological identification of neurons that contain this enzyme has been difficult in mammals. Several aromatase enzyme antisera have been developed and used in brain, and each gives a different pattern of immunoreactivity. Moreover, despite the fact that aromatase activity is very high in the bed nucleus of the stria terminalis, several amygdala nuclei, the preoptic area and hypothalamus, AROM-ir in these regions has been very limited. The distribution of AROM-ir in female musk shew brain tissues is modified by treatment with vorozole prior to sacrifice. Female musk shrew brains contain aromatase immunoreactive cell bodies, as reported previously, in the central amygdala, lateral septum and to a limited extent in the bed nucleus of the stria terminalis (BST). Brains of females treated with vorozole show additional immunoreactivity in the preoptic area, hypothalamus, and medial amygdala, and have a broad distribution of AROM-ir in several subdivisions of the BST. Several sexual dimorphisms are apparent in musk shrews brains after treatment with vorozole. We have quantified this sexual dimorphism in the medial preoptic area (MPO) by counting immunoreactive cells. In both the rostral and caudal portions of the MPO, female brains contain significantly fewer AROM-ir cell bodies than males. These data are in complete agreement with sex differences in biochemical analyses of aromatase activity in the MPO. At this time we do not know if these dimorphisms are the result of differences in circulating levels of steroids in males and females, and/or if the AROM-ir nuclei regulate sexually dimorphic behaviors. PMID- 8730655 TI - Ontogeny of pituitary transcription factor-1 (Pit-1), growth hormone (GH) and prolactin (PRL) mRNA levels in male and female rats and the differential expression of Pit-1 in lactotrophs and somatotrophs. AB - Pituitary transcription factor-1 (Pit-1 or GHF-1), a transcription factor specific to the anterior pituitary, is involved in the expression and regulation of the growth hormone (GH) and prolactin (PRL) genes. Post-pubertally, the expression of both GH and PRL becomes sexually dimorphic with males having higher GH levels and females higher PRL levels; however, little is known about the postnatal regulation of their common transcription factor. Furthermore, whether the Pit-1 gene is differentially expressed in somatotrophs and lactotrophs remains to be elucidated. In this study, we used in situ hybridization histochemistry to examine Pit-1, GH and PRL mRNA levels in the anterior pituitaries of male and female rats throughout development (0, 5, 10, 20, 30, 40 and 60 days of age) to determine when GH and PRL production becomes sexually dimorphic and if this is accompanied by a dimorphism in Pit-1 gene expression. In addition, the level of Pit-1 mRNA was determined separately in both GH mRNA and PRL mRNA containing cells during the various developmental stages. We found that in both males and females the mRNA levels of Pit-1, GH and PRL remain relatively unchanged until around the time of pubertal onset (30-40 days) when there is a significant increase in all three mRNA species, which is followed by a decrease to adult levels. Also around the time of puberty, both GH and PRL mRNA levels become sexually dimorphic, with males having higher levels of GH mRNA and females higher PRL mRNA levels. In contrast, at no time during development were overall Pit-1 mRNA levels found to differ between the sexes. However, when Pit-1 mRNA content was measured separately in specific cell types, significant differences between the sexes became evident. Throughout development Pit-1 mRNA levels are higher in lactotrophs of females than in those of males, whereas in somatotrophs males have higher Pit-1 mRNA levels than females. Furthermore, within a sex there is differential expression of Pit-1 in the two cell types with females having significantly higher levels of Pit-1 in lactotrophs than in somatotrophs and males having higher levels in somatotrophs than in lactotrophs. These data support the hypothesis that a sexual dimorphism exists in the expression and pituitary specific transcription factor Pit-1; however, this dimorphism is not manifest as a difference in overall mRNA levels, but in the differential expression of this gene in lactotrophs and somatotrophs. PMID- 8730656 TI - Oxytocin released within the supraoptic nucleus of the rat brain by positive feedback action is involved in parturition-related events. AB - Oxytocin is released within the supraoptic nucleus during parturition and suckling. During suckling, such release is important in positive feedback stimulation of oxytocin neurons. We have investigated whether oxytocin released within this hypothalamic nucleus during parturition (1) acts on local receptors to further amplify its own release in a positive feedback manner and (2) is critically involved in the regulation of the delivery process. To examine the effect of the oxytocin antagonist on oxytocin release within the supraoptic nucleus, microdialysates were sampled before and during parturition and either vehicle or the antagonist was infused adjacent to the microdialysis probe directly into the supraoptic nucleus after delivery of the second pup. Intranuclear infusion of an oxytocin receptor antagonist (des-Gly NH2d(CH2)5[Tyr(Me)2Thr4]OVT; 50 ng/0.5 microliters) significantly (P < 0.01) diminished the parturition-related rise in oxytocin release within the supraoptic nucleus and reduced the number of pups delivered during the first and second 30 min dialysis period compared to vehicle-treated controls. Bilateral infusion of the oxytocin receptor antagonist into the supraoptic nucleus after delivery of the second pup significantly slowed parturition (P < 0.05), although the parturition-related rise in plasma oxytocin concentration was unchanged. In addition, the onset of suckling was significantly affected by the antagonist as indicated by fewer liver pups and fewer surviving pups with milk in their stomachs 24 hours after parturition (P < 0.05). To seek other, periventricular sites of oxytocin action during parturition, oxytocin or the oxytocin antagonist was infused into the lateral cerebral ventricle from the birth of pup 2. Via this route, oxytocin speeded up parturition, but the antagonist was ineffective; thus it appears that periventricular oxytocin-sensitive sites are not normally active in promoting parturition, and can do so. The findings indicate a receptor mediated positive feedback action of oxytocin on its own release within the supraoptic nucleus during parturition, which seems to be involved in the progress of parturition without significantly affecting circulation oxytocin levels. Oxytocin released within the supraoptic nucleus might be important for the coordinated activation of oxytocin neurons and for the synergistic central and peripheral oxytocin effects involved in the regulation of parturition-related events necessary for the survival of the newborn, including the onset of lactation. PMID- 8730657 TI - Nitric oxide induces morphological changes in cultured neurohypophysial astrocytes. AB - Cultured pituicytes, derived from the neurohypophysis of adult rats, have previously been reported to change from a non-stellate form to a stellate form when incubated in medium containing a beta-adrenoreceptor agonist. This study was designed to determine whether the same morphological change could be induced by direct activation of adenylate cyclase or of soluble guanylate cyclase. The fraction of stellate cells was normally low (< 0.25) when the pituicytes were incubated (90 min) in a HEPES buffered salt solution (HBSS); most pituicytes had an amorphous protoplasmic appearance. The fraction of stellate cells was significantly increased when pituicytes were incubated in HBSS supplemented with isoproterenol (10 microM) or forskolin (5 microM) or with either of the nitric oxide donors nitroprusside (10-25 microM) and 3-morpholinosydnonimine (SIN-1; 10 microM). The effect of forskolin was mimicked by 8-bromo cyclic AMP, a membrane permeable analog of cyclic AMP, but not by the inactive forskolin analog 1, 9 dideoxyforskolin. The effect of nitroprusside was blocked by methylene blue, an inhibitor of soluble guanylate cyclase, and was mimicked by 8-bromo cyclic GMP, a membrane permeable analog of cyclic GMP. These results demonstrate that activation of adenylate cyclase and also of soluble guanylate cyclase can induce pituicytes to undergo morphological changes in vitro. The data suggest that the activity of both enzymes may be important in control of the plastic relationship that exists between neuronal and glial elements in the neurohypophysis in vivo. PMID- 8730658 TI - Integrating sphere effect in whole-bladder wall photodynamic therapy: III. Fluence multiplication, optical penetration and light distribution with an eccentric source for human bladder optical properties. AB - Whole-bladder-wall (WBW) photodynamic therapy (PDT) is performed using approximately 630 nm light emitted by an isotropic light source centered in the bladder cavity. The phenomenon of an increased fluence rate in this spherical geometry, due to light scattering, is denoted as the integrating sphere effect. The fluence rate and the optical penetration depth depend on a single tissue optical parameter, namely the reduced albedo. The optical properties of (diseased) human bladder tissue, i.e. absorption coefficient, scattering coefficient, anisotropy factor and refractive index, were determined in vitro in the wavelength range of 450-880 nm. The integrating sphere effect and optical penetration depth were calculated with diffusion theory and compared to Monte Carlo (MC) computer simulations using approximately 630 nm optical properties. With increasing albedo, the integrating sphere effect calculated with diffusion approximation is increasingly larger than that found with MC simulations. Calculated and simulated optical penetration depths are in reasonable agreement. The smaller the integrating sphere effect for a given tissue absorption, the larger the optical penetration depth into the bladder wall, as the effective attenuation coefficient decreases. Optical penetration depths up to approximately 7.5 mm (definition dependent) can be responsible for unintended tissue damage beyond the bladder tissue. MC simulations were also performed with an eccentric light source and the uniformity of the light distribution at the bladder wall was assessed. The simulations show that even for a small eccentricity, the extremes in deviation from the mean fluence rate are large. All these results indicate that WBW PDT should be performed with some kind of in situ light dosimetry. PMID- 8730659 TI - A study of various parameters of spherically curved phased arrays for noninvasive ultrasound surgery. AB - The spherically curved square element phased array design for ultrasound surgery was studied in several of its important aspects. A method for determining the spatial limitations of the multiple-foci region was developed for spherically curved phased arrays. The effects on the ultrasound fields of varying the phases and the amplitudes at control points were investigated. It was found that the phases and the magnitudes of control points have an impact on the ultrasound field distributions. The effects of multiple-foci spacing, pulse duration, and maximum temperature on the size and shape of necrosed tissue were investigated. For a spherically curved phased array with an 8 cm radius of curvature, an 8.8 x 8.8 cm2 projected area, and a 1.5 MHz operating frequency, the minimum number of phased array elements that could produce the largest acceptable necrosed tissue volume was 256. The tissue volume necrosed during the sonication could be increased to approximately 1 x 1 x 3 cm3 with this array. PMID- 8730660 TI - Wall effects in plane-parallel ionization chambers. AB - All dosimetry protocols recommend the use of plane-parallel chambers for dose determination in electron beams with energies below 10-15 MeV. The protocols have assumed chamber perturbation effects to be negligible. The new AAPM Protocol (TG39) includes a cavity replacement factor prepl that differs from unity for some chambers, but assumes that the wall perturbation factor, pwall, may be taken as unity. In this paper the perturbation of the wall has been determined, using a large plane-parallel ionization chamber with exchangeable front and back walls. The results show that in many commercial chambers there is an energy dependent pwall factor, mainly due to differences in backscatter from the often thick chamber body as compared to the phantom material. Backscatter in common phantom and chamber materials may differ by as much as 2% at low electron energies (Plastic Water as compared to polystyrene). The front walls are often thin, resulting in negligible perturbation, but the 0.5 mm front wall of graphite in the NACP chamber was found to increase the response by 0.7% in a PMMA phantom. The experimental results have been compared with EGS4 Monte Carlo calculations using the DOSRZ code. There is an agreement within statistical and experimental uncertainties (0.5%) indicating that it is possible to use Monte Carlo calculations to calculate perturbation factors with good accuracy. PMID- 8730661 TI - 10 MV x-ray SMRs obtained using zero-area S(p) correction factors derived by means of the Bjarngard-Petti method. AB - Based on the Khan concept, a new set of 10 MV x-ray scatter-maximum ratios (SMRs) has been produced. THese new SMRs reflect no electron contamination and are parametrized. A method has been developed for determining the minimum field size with lateral CPE (charged particle equilibrium) at the field centre. It is found that for 10 MV x-rays the minimum square field with lateral CPE at large depths on the central axis in water is 3.5 x 3.5 cm2. The zero-area phantom scatter correction factor (S(p)) is derived using the Bjarngard-Petti method. It is found that the effect of electron contamination is to reduce the zero-area Sp correction factor. PMID- 8730662 TI - Calculation of the absorbed dose distribution due to irregularly shaped photon beams using pencil beam kernels derived form basic beam data. AB - In radiotherapy, accurately calculated dose distributions of irregularly shaped photon beams are needed. In this paper, an algorithm is presented which enables the calculation of dose distributions due to irregular fields using pencil beam kernels derived from simple basic beam data usually measured on treatment units, i.e. central axis depth-dose curves and profiles. The only extra data that are needed, and are not currently measured, is the phantom scatter factor curve at the reference depth. The algorithm has been developed as an extension to a previously developed algorithm for rectangular fields which is based on the Milan Bentley storage model. In the case of an irregular field, the depth dose and the boundary function are computed by convolution of a field intensity function with pencil beam kernels. The depth dose is computed by using a 'scatter' kernel, which is derived from the stored depth-dose curves and from the phantom scatter factor curve. The boundary function is computed by using a 'boundary' kernel, which is derived from the boundary profile of a number of large square fields. Because of the simplicity of the data used and the underlying concepts, which for instance do not separate the head scatter from the primary beam, this algorithm presents some shortcomings. On the other hand, this simplicity is also of great advantage and the inaccuracy is acceptable for most clinical situations. PMID- 8730663 TI - Characteristics of the absorbed dose to water standard at ENEA. AB - The primary standard of absorbed dose to water established at ENEA for the Co-60 gamma-ray quality is based on a graphite calorimeter and an ionometric transfer system. This standard was recently improved after a more accurate assessment of some perturbation effects in the calorimeter and a modification of the water phantom shape and size. The conversion procedure requires two corresponding depths, one in graphite and one in water, where the radiation energy spectra must be the same. The energy spectra at the corresponding points were determined by a Monte Carlo simulation in water and graphite scaled phantoms. A thorough study of the calorimeter gap effect corrections was also made with regard to their dependence on depth and field size. A comparison between the ionization chamber calibration procedures based on the standards of absorbed dose to water and of air kerma was also made, confirming the consistency of the two methods. PMID- 8730664 TI - Computer-aided design optimization with the use of a fast dose model for linear accelerator-based stereotactic radiosurgery. AB - In order to efficiently plan non-spherical radiosurgical targets we have used computer-aided design optimization techniques with a fast dose model. A study of the spatial dose distribution for single or multiple non-coplanar arcs was carried out using a 18 cm diameter spherical head model. The dose distribution generated from the 3D dose computation algorithm can be represented by a simple analytic form. Two analytic dose models were developed to represent the dose for preset multiple non-coplanar arcs or a single arc: spherical and cylindrical. The spherical and cylindrical dose models compute dose quickly for each isocentre and single arc. Our approach then utilizes a computer-aided design optimization (CAD) with the use of two fast approximate dose models to determine the positions of isocentres and arcs. The implementation of CAD with fast dose models was demonstrated. While the fast dose models are only approximations of the true dose distribution, it is shown that this approximate model is sufficient to optimize isocentric position, collimator size and arc positions with CAD. PMID- 8730665 TI - A portal image alignment and patient setup verification procedure using moments and correlation techniques. AB - The objective of this study was to develop an integrated field shape and patient setup verification procedure for portal images. The proposed procedure used one of the approved portal images as the reference image for automated comparison with subsequent portal images. The first step of the procedure used a moments method to align treatment field boundaries. This step was necessary to compensate for the repositioning error of an image detector and to create a common frame of reference for comparing anatomical shifts relative to the field boundary. At the end of the moments alignment, a moments figure of merit was computed and compared with a pre-established threshold. This verified whether there was a potential shape change in the treatment field. To measure anatomical misalignment, the last step in the procedure was to use a grey-scale image correlation method to align translations and in-plane rotations relative to the anatomy of the reference image. The procedure was shown in phantom studies to err by less than 1 mm when detecting translational shifts and less than 1 degree when detection in-plane rotations. The moments verification method showed a sensitivity of detecting a placement error of 6 mm for a single leaf in a controlled experiment where a multileaf collimator was used for field shaping. The alignment procedure was fast and could be done in less than 12 s on an IBM-compatible 486 personal computer. PMID- 8730666 TI - Numerical solutions of differential equations of a cylindrical ionization chamber. AB - The differential equations of the cylindrical ionization chamber filled with air have been solved numerically for different ratios of the outer and inner electrode for both polarities of the voltage supply. The obtained distributions of the electric field strength inside the chamber confirm the analysis of Thomson and Thomson and the calculations made by Mie. The calculated collection efficiencies are in good agreement with the results of the calculation of Sprinkle and Tate, and with the experimental data of Moriuchi et al. PMID- 8730667 TI - The measurement of the velocity of ultrasound in fixed trabecular bone using broadband pulses and single-frequency tone bursts. AB - The velocity of ultrasound in a series of 10 fixed os calces was measured using both short pulses and 750 kHz tonebursts. The values obtained from the pulse measurements differed from the toneburst values by up to 16% depending on the selection of the zero-crossing point used as a reference in the pulse measurements. It is demonstrated that the discrepancy between the values is itself a function of the frequency-dependent attenuation in the propagating medium and this is confirmed by theoretical modelling. The toneburst results are also compared with measurements using a cross-correlation technique, and these show a close agreement. PMID- 8730668 TI - Optimization of factors affecting the state of normality of a medical image. AB - The purpose of this paper is to examine the first stage of the diagnostic process in medical imaging, namely determination of the state of normality, and to attempt to optimize factors contributing to this stage. An image of a given type is defined as abnormal if it does not belong to the appropriate class of normal images. All images must be pre-processed involving image registration and normalization to align and scale the images with respect to each other. Normal ranges may be determined for each voxel (or other appropriate region) from a representative normal sample using univariate analysis, obtaining mean and standard deviation images, or multivariate analysis, which accounts also for normal patterns of variation (represented as principal components). For a new image, the variation from normality (in SDs) for each region may be determined. Since the spatial distribution of this parameter is thought to be relevant, connectivity of abnormal voxels was considered as a possible factor. For the purposes of this study, SPECT images indicating regional cerebral blood flow were used. Images from 50 normal subjects formed the normal sample. A further 40 normal subjects and 200 patients referred with suspected dementia were then analyzed using the normal ranges. ROC analysis, using number of SDs as a variable threshold, was used to optimize the factors. Normalization to global values followed by multivariate analysis using four or five principal components provided optimal discrimination. Connectivity of voxels emerged as an important factor, around 10 connected voxels being optimal for this study. PMID- 8730669 TI - An investigation of light transport through scattering bodies with non-scattering regions. AB - Near-infra-red (NIR) spectroscopy is increasingly being used for monitoring cerebral oxygenation and haemodynamics. One current concern is the effect of the clear cerebrospinal fluid upon the distribution of light in the head. There are difficulties in modelling clear layers in scattering systems. The Monte Carlo model should handle clear regions accurately, but is too slow to be used for realistic geometries. The diffusion equation can be solved quickly for realistic geometries, but is only valid in scattering regions. In this paper we describe experiments carried out on a solid slab phantom to investigate the effect of clear regions. The experimental results were compared with the different models of light propagation. We found that the presence of a clear layer had a significant effect upon the light distribution, which was modelled correctly by Monte Carlo techniques, but not by diffusion theory. A novel approach to calculating the light transport was developed, using diffusion theory to analyze the scattering regions combined with a radiosity approach to analyze the propagation through the clear region. Results from this approach were found to agree with both the Monte Carlo and experimental data. PMID- 8730671 TI - European ministers consider options for resolving the beef crisis. PMID- 8730670 TI - Comments on 'A new method to determine ratios of electron stopping powers to an improved accuracy'. PMID- 8730672 TI - Assessment of the effect of three treatments to remove intrauterine fluid on pregnancy rate in the mare. AB - The effects on pregnancy rate of three different treatments to remove intrauterine fluid were assessed in 1267 mares. The mares were mated and allocated, in strict rotation, to four treatment groups: 1) untreated controls, 2) intrauterine infusion of broad spectrum antibiotics, 3) intravenous injection of oxytocin, 4) intravenous injection of oxytocin followed by intrauterine antibiotics. The pregnancy status of the mares was determined 13 to 15 days and 27 to 30 days after ovulation by transrectal ultrasonography. The pregnancy rate of group 4 (72 per cent) was higher than that of group 2 (64 per cent, P < 0.01) or group 3 (63 per cent, P < 0.01). The pregnancy rates of groups 2 and 3 were higher than that of group 1 (56 per cent, P < 0.01). The treatment with antibiotics and oxytocin appeared to have an additive beneficial effect which suggested two different modes of action of the combination treatment, namely antibacterial activity and fluid drainage. In the untreated mares more fluid accumulated in the uterine lumen after mating, and this was the most likely reason for their lower pregnancy rate. PMID- 8730673 TI - Clinical and pathological features of Nigerian equine encephalitis. AB - Thirteen cases of a disease with a low morbidity and very high mortality in horses in Nigeria are described; the disease is characterised by fever (rectal temperature > or = 40 degrees C), generalised muscle spasms, ataxia, increased respiratory and heart rates and terminal lateral recumbency. The illness generally lasts three to five days but durations of 12 to 30 hours have been observed. Laboratory investigations, including histopathology and serology suggest a viral aetiology, possibly an alphavirus of the equine encephalitis group. PMID- 8730674 TI - Sulphur-induced polioencephalomalacia in lambs. AB - An outbreak of polioencephalomalacia affected 16 of 46 Swaledale lambs and five of 25 Scottish blackface lambs 15 to 32 days after they were introduced to an ad libitum concentrate ration containing 0.43 per cent sulphur. The clinical signs were acute and included depression central blindness and head-pressing, but no hyperaesthesia, nystagmus, dorsiflexion of the neck or opisthotonos were observed. Treatment of the affected lambs with vitamin B1, dexamethasone and antibiotics was associated with a prolonged recovery period, though no further cases were identified after vitamin B1 had been given parenterally to all the lambs at risk. PMID- 8730675 TI - Efficacy of a phyto-aromatic gel against auricular manage in rabbits and carnivores. AB - The efficacy of a phyto-aromatic gel (Canidor) was evaluated, in vitro against Psoroptes cuniculi and in vivo against P cuniculi and Otodects cynotis, in experimentally and naturally infested rabbits and domestic carnivores respectively. At a 1/1 dilution the gel was 100 per cent active in vitro against P cuniculi after less than six hours of contact. In experimentally infested rabbits the gel was administered daily during two periods of five consecutive days, six days apart. A clinical cure was achieved in all the animals, but two harboured a few eggs and mites at the end of the trial. In dogs and cats the active formulation was given for two periods of four consecutive days, seven days apart. Clinical and parasitological examinations 10 and 30 days after initiating the treatment regiment revealed an excellent acaricidal effect. PMID- 8730676 TI - Comparison of ELISA and selective culture in the diagnosis of swine dysentery in Thailand. PMID- 8730677 TI - Parathyroid hormone-related protein in the colostrum of paretic post parturient dairy cows. PMID- 8730678 TI - Effect of vaccination against Leptospira interrogans serovar hardjo on milk production and fertility in dairy cattle. PMID- 8730679 TI - Lead concentration in blood, milk and feed of lactating buffalo after acute lead poisoning. PMID- 8730680 TI - Rare breeds and the BSE situation. PMID- 8730681 TI - Transport of pigs. PMID- 8730682 TI - Haemolytic anaemia in an embryo transfer calf. PMID- 8730683 TI - Milford Haven oil spill. PMID- 8730684 TI - Surgical management of canine cervical spondylopathy. PMID- 8730685 TI - Pulse oximetry for assessing oxygen status of neonatal calves. PMID- 8730686 TI - Government considers cattle cull to restore confidence in beef. PMID- 8730687 TI - Government announcement sparks a crisis for British beef. PMID- 8730688 TI - Statement issued by the spongiform encephalopathy advisory committee on March 24. PMID- 8730689 TI - Ageing horses by an examination of their incisor teeth: an (im)possible task? AB - It is generally considered that the age of a horse can be determined by examining its incisor teeth. However, the criteria used to determine age from dental configurations differ widely. The existence of this variety of rules and guidelines was the challenge for the present examination. Detailed descriptions of the incisor teeth of 212 horses of registered age were recorded and the results were compared with the age criteria of various authors. The time at which teeth were shed and the appearance of dental stars seemed to be more reliable features than the disappearance of the cups. The disappearance of the marks occurred four years later than usually stated. Other criteria, such as the presence of the seven-year notch and Galvayne's groove, were too variable and inconsistent to be reliable for the determination of age. PMID- 8730690 TI - Deposition in the distal parts of the bovine respiratory tract: assessment of equipment suitable for drug inhalation. AB - The efficiency of equipment suitable for the inhalation of drugs by calves was assessed in six animals which inhaled radioisotopically labelled particles while suffering from reversible diffuse bronchoconstriction induced experimentally with 5-hydroxytryptamine and while they were breathing normally. Respiratory rates and data from pulmonary function tests and scintiscans were recorded during both investigations. After the first investigation, a mean (se) wash-out period of 9.8 (3.2) days was allowed. Under diffuse bronchoconstriction, the respiratory rate, the oscillatory resistance and the compliance of the respiratory system reached 282.1 (22.0), 161.1 (10.8) and 68.8 (2.7) per cent of their respective baseline values. When the calves were breathing normally these parameters did not change over time. The ratios (Cp/Ct) of the counts of gamma-disintegrations in the peripheral part (Cp) of the lungs and in the total lung area (Ct) were not significantly different when comparing the results from the two investigations. The ratios of Cp/Ct in the left lungs did not differ significantly from those in the right lungs. PMID- 8730691 TI - Low bulk milk somatic cell counts and endotoxin-associated (toxic) mastitis. PMID- 8730692 TI - Erythromycin-resistant thermophilic Campylobacter species isolated from pigs. PMID- 8730693 TI - Self destructive behaviour in dairy cattle. PMID- 8730695 TI - TB testing. PMID- 8730694 TI - Meat hygiene service. PMID- 8730696 TI - Relative risk. PMID- 8730697 TI - Escherichia coli O157 infection in a human linked to exposure to infected livestock. PMID- 8730698 TI - Wildlife rehabilitation. PMID- 8730699 TI - Wild Mammals (Protection) Act. PMID- 8730700 TI - Mange in domesticated rats. PMID- 8730701 TI - Protein kinase C activation is necessary but not sufficient for induction of long term potentiation at the synapse of mossy fiber-CA3 in the rat hippocampus. AB - The involvement of protein kinase C in long-term potentiation was investigated in the mossy fiber-CA3 pathway in an in vitro slice preparation of rat hippocampus. Tetanic stimulation induced stable long-term potentiation in the mossy fiber-CA3 pathway which was not affected by N-methyl-D-aspartate receptor antagonists. Long term potentiation was not induced in the presence of a protein kinase C inhibitor, sphingosine. Application of 1 microM phorbol-12, 13-diacetate, an activator of protein kinase C, potentiated the synaptic response by about 400% and this potentiation was completely reversible upon washing. Sphingosine blocked the potentiation when it was applied before protein kinase C activation by phorbol-12, 13-diacetate. However, sphingosine had no effect on the potentiation when it was applied after the synaptic response was potentiated to a plateau following phorbol-12,13-diacetate perfusion. Long-term potentiation and phorbol ester-induced potentiation were not additive when phorbol-12,13-diacetate was applied after induction of long-term potentiation, suggesting that long-term potentiation and phorbol-12, 13-diacetate activate the same protein kinase C pool. The enhanced response caused by phorbol-12,13-diacetate returned to the long-term potentiation level after wash-out of phorbol-12,13-diacetate. Thus the cellular changes underlying long-term potentiation are long-lasting or permanent, while those caused by phorbol-12,13-diacetate are not. However, if tetanic stimulation was induced during prolonged phorbol-12,13-diacetate application (1 h), a potentiation similar in amplitude to long-term potentiation was induced but the population response returned to the control pre-long-term potentiation level after 2 h of washing. The potentiation following tetanic stimulation during prolonged application of phorbol-12,13-diacetate was blocked in the presence of D 2-amino-5-phosphonovaleric acid, a N-methyl-D-aspartate receptor antagonist. Thus, in the presence of phorbol esters the N-methyl-D-aspartate-independent long term potentiation is occluded but a transient potentiation appears, presumably due to hyperexcitability and activation of N-methyl-D-aspartate receptors in recurrent pathways of area CA3. Normal N-methyl-D-aspartate-independent long-term potentiation could be induced after the 2 h washout period and now was maintained. In conclusion, protein kinase C activation is essential but not sufficient for long-term potentiation in the mossy fiber-CA3 pathway and when stimulated by application of phorbol esters produces a large and reversible synaptic potentiation. These investigations show that long-term potentiation in CA3 is a complex event involving several steps, and that activation of protein kinase C is only one of them. PMID- 8730702 TI - Distribution of AMPA receptor subunits in the hippocampal formation of temporal lobe epilepsy patients. AB - The immunocytochemical distribution of the AMPA-selective receptor subunits GluR1 and GluR2/3 were mapped in the human hippocampal formation obtained from surgery for medically intractable temporal lobe epilepsy. GluR2/3 immunoreactivity was detected in all principal cell types of the hippocampal formation, including hilar neurons, granule cells of the dentate gyrus, and pyramidal cells of the cornu ammonis fields and subiculum. GluR2/3 immunostaining typically filled the cell bodies and processes of neurons. A comparison of GluR2/3 immunoreactivity in a sclerotic specimen versus a non-sclerotic specimen demonstrated a profound loss of staining, specifically in the areas where neuronal dropout was occurring, including CA1, CA3 and the hilus. An analysis of GluR1 immunoreactivity in non sclerotic specimens revealed that it was predominantly localized to cellular processes throughout the cornu ammonis fields, with a sparse staining of the dentate gyrus outer molecular layer and little to no staining of the dentate gyrus inner molecular layer. Similar to the GluR2/3-immunostained patterns, GluR1 immunoreactivity was lost in the cornu ammonis fields of sclerotic hippocampal specimens, corresponding to patterns of neuronal dropout. Our most compelling finding was a unique extensive pattern of GluR1 and Glu2/3 immunoreactivity throughout the molecular layers of the dentate gyrus of severely compromised hippocampi. The altered staining of GluR1 and GluR2/3 complements some of the patterns of axonal sprouting already described for the dentate gyrus, with a conjecture that their anatomy and distribution pattern underlies to some degree the reorganization of the sclerotic hippocampus. A combination of enhanced glutamatergic transmission and changes in neuropeptides that modulate hippocampal circuitry could greatly affect the degree of excitability in the hippocampal formation. The alterations of GluR1 and GluR2/3 immunoreactivity in the dentate gyrus add another component to the concept of reorganization in the epileptic sclerotic hippocampus. PMID- 8730703 TI - Extracellular slow negative transient in the dentate gyrus of human epileptic hippocampus in vitro. AB - We investigated extracellular slow negative transient in dentate granule cells of human epileptic hippocampus. Hippocampal slices were prepared from brain specimens removed from 23 patients who underwent surgical treatment for medically intractable seizures. In 15 patients, hippocampi were sclerotic and the aberrant anatomical reorganization of dentate granule cell axons (mossy fibers) was detected. In eight patients, hippocampi were non-sclerotic and nominal reorganization was detected. Single perforant path stimulation evoked field responses in dentate granule cells in all 23 hippocampi. In sclerotic hippocampi, evoked field responses were followed by a slow onset extracellularly negative potential, which appeared gradually in the course of low frequency stimulation of the perforant path. Single action potentials could be recorded from negative potentials indicating that these potentials represented dentate granule cell depolarization. A low concentration of bicuculline methiodide (10 microM), a GABAA receptor antagonist, facilitated the appearance of negative potentials suggesting that a reduction in functional inhibition could unmask these potentials. The application of D-2 amino-5-phosphonovaleric acid blocked extracellular negative potentials, but initial perforant path responses were spared. This finding suggested that negative potentials were at least in part mediated by the N-methyl-D-aspartate receptor subtypes in their generation. In contrast, in non-sclerotic hippocampi with nominal "reorganization", no extracellular negative potentials were observed. The present study suggests that dentate granule cell excitability could be amplified when their reorganized axonal pathways were present in human sclerotic hippocampus as previously proposed in animal models of epilepsy. PMID- 8730704 TI - Hippocampal activity during transient respiratory events in the freely behaving cat. AB - We measured dorsal hippocampal activity accompanying sighs and apnea using reflectance imaging and electrophysiologic measures in freely behaving cats. Reflected 660-nm light from a 1-mm2 area of CA1 was captured during sighs and apnea at 25 Hz through a coherent image conduit coupled to a charge coupled device camera. Sighs and apnea frequently coincided with state transitions. Thus, state transitions without apnea or sighs were separately assessed to control for state-related activity changes. All dorsal hippocampal sites showed discrete regions of activation and inactivation during transient respiratory events. Imaged hippocampal activity increased 1-3 s before the enhanced inspiratory effort associated with sighs, and before resumption of breathing after apnea. State transitions lacking sighs and apnea did not elicit analogous optical activity patterns. The suprasylvian cortex, a control for site, showed no significant overall reflectance changes during phasic respiratory events, and no discrete regions of activation or inactivation. Spectral estimates of hippocampal electroencephalographic activity from 0-12 Hz showed significantly increased power at 3-4 Hz rhythmical slow activity before sighs and apnea, and increased 5 6 Hz rhythmical slow activity power during apnea, before resumption of breathing. Imaged activity and broadband hippocampal electroencephalogram power decreased during sighs. We propose that increased hippocampal activity before sigh onset and apnea termination indicates a role for the hippocampus in initiating inspiratory effort during transient respiratory events. PMID- 8730705 TI - Differential expression of NADPH diaphorase in functionally distinct prefrontal cortices in the rhesus monkey. AB - The prefrontal cortex of primates is an integrative centre for sensory, cognitive, mnemonic and emotional processes. The cellular features which contribute to the functional specialization of its subsectors are poorly understood. In this study we determined the distribution of nicotinamide adenine dinucleotide phosphate-diaphorase-positive neurons in structurally and functionally distinct prefrontal cortices in the rhesus monkey. This class of neurons express nitric oxide synthase which is necessary for the production of nitric oxide, a novel neural messenger implicated in learning and memory. The density of diaphorase-positive neurons was approximately four times higher in olfactory areas than in eulaminate areas (areas 9, 10, 12, 46, and 8), and two- to three-times higher in the agranular limbic area PAll than in eulaminate areas. Positive neurons were concentrated in a deep band (layers V and VI), a superficial band (layers II and upper III), and were sparsely distributed in the central, thalamic recipient zone (deep layer III, layer IV and upper V). The highest densities of positive neurons were observed in the white matter where their prevalence followed the opposite trend than in the corresponding overlying cortices. The distribution of diaphorase-positive neurons was correlated with the regional anatomic and functional specialization of prefrontal cortices. Thus, diaphorase-positive neurons were most densely distributed in orbital and then medial prefrontal limbic cortices which have a low cell density and widespread connections. In contrast, positive neurons were comparatively sparse in eulaminate cortices, which have a high cell density and more restricted connections. These findings indicated that the distribution of diaphorase positive neurons in prefrontal cortices is not random, but is associated with the structural architecture and functional attributes of these cortices. The preponderance of diaphorase-positive neurons in limbic cortices, which have been implicated in learning and memory, is consistent with the idea that nitric oxide may have a role in synaptic plasticity. PMID- 8730706 TI - Intrastriatal implants of polymer encapsulated cells genetically modified to secrete human nerve growth factor: trophic effects upon cholinergic and noncholinergic striatal neurons. AB - Nerve growth factor selectively prevents the degeneration of cholinergic neurons following intrastriatal infusion but rescues both cholinergic and noncholinergic striatal neurons if the nerve growth factor is secreted from grafts of genetically modified fibroblasts. The present study evaluated whether grafted fibroblasts genetically modified to secrete human nerve growth factor could provide trophic influences upon intact cholinergic and noncholinergic striatal neurons. Unilateral striatal grafts of polymer-encapsulated cells genetically modified to secrete human nerve growth factor induced hypertrophy and significantly increased the optical density of choline acetyltransferase immunoreactive striatal neurons one, two, and four weeks post-transplantation relative to rats receiving identical grafts missing only the human nerve growth factor construct. Nerve growth factor secreting grafts also induced a hypertrophy of noncholinergic neuropeptide Y-immunoreactive striatal neurons one, two, and four weeks post-transplantation. Glutamic acid decarboxylase-immunoreactive neurons were unaffected by the human nerve growth factors secreting grafts. The effects upon choline acetyltransferase-immunoreactive and neuropeptide Y immunoreactive striatal neurons dissipated following retrieval of the implants. Immunocytochemistry for nerve growth factor revealed intense graft-derived immunoreactivity for up to 1000 microns from the capsule extending along the dorsoventral axis of the striatum. Nerve growth factor-immunoreactivity was also observed within a subpopulation of striatal neurons and may represent nerve growth factor consumer neurons which retrogradely transported graft-derived nerve growth factor. When explanted, grafts produced 2-4 ng human nerve growth factor/24 h over the time course of this study indicating that this level of continuous human nerve growth factor secretion was sufficient to mediate the effects presently observed. PMID- 8730707 TI - Functional neuroanatomy of the nigrostriatal and striatonigral pathways as studied with dual probe microdialysis in the awake rat--I. Effects of perfusion with tetrodotoxin and low-calcium medium. AB - In the present study we employed the dual probe approach to investigate functional interactions between the nigrostriatal dopaminergic and striatonigral GABAergic pathways in the awake, freely moving rat and their role in motor function. One microdialysis probe of concentric design was implanted in the substantia nigra pars reticulata and another in the ipsilateral dorsolateral striatum. Perfusion with a low-Ca2+ (0.1 mM) medium and with the voltage dependent Na(+)-channel blocker tetrodotoxin (10 microM) was alternatively performed in both brain regions and the dialysate dopamine, glutamate and GABA levels were simultaneously measured in the dorsolateral striatum, whereas GABA levels alone were monitored in the substantia nigra. Perfusion with a low-Ca2+ medium in the substantia nigra pars reticulata did not affect local GABA levels, but transiently increased striatal dopamine release (+40%) without modifying striatal glutamate and GABA levels. Conversely, intranigral perfusion with tetrodotoxin transiently increased local GABA levels (+40%), while it decreased striatal dopamine (-60%) and increased glutamate (+70%) and GABA (+50%) levels. Perfusion with a low-Ca2+ medium in the dorsolateral striatum reversibly decreased local dopamine (-70%), glutamate (-20%) and GABA (-20%) levels, while local perfusion with tetrodotoxin decreased dopamine (-70%), increased glutamate (+30%) but did not affect dialysate GABA levels in this brain area. Neither of these intrastriatal treatments significantly affected GABA levels in the substantia nigra. Intranigral but not intrastriatal perfusion with tetrodotoxin was also associated with an increase in spontaneous locomotor activity as expressed by contralateral turning. Intranigral and intrastriatal perfusion with low-Ca2+ medium did not influence locomotor activity. On the basis of these neurochemical and behavioural findings, we propose a new dynamic model for the study of motor behaviour as mediated by basal ganglia circuitry. PMID- 8730708 TI - Functional neuroanatomy of the nigrostriatal and striatonigral pathways as studied with dual probe microdialysis in the awake rat--II. Evidence for striatal N-methyl-D-aspartate receptor regulation of striatonigral GABAergic transmission and motor function. AB - In the present study we used the dual probe approach to investigate striatal N methyl-D-aspartate receptor regulation of GABA release from the substantia nigra pars reticulata of the awake, freely moving rat. One microdialysis probe of concentric design was implanted in the dorsolateral striatum and another in the ipsilateral substantia nigra pars reticulata. Perfusion with N-methyl-D-aspartate (100 microM) in the dorsolateral striatum decreased local dopamine release (-25%) and increased both glutamate (+40%) and GABA (+35%) release. Moreover, perfusion with N-methyl-D-aspartate (100 microM) in the dorsolateral striatum increased GABA release (+20%) in the substantia nigra pars reticulata. Perfusion with the lower (10 microM) N-methyl-D-aspartate concentration in the dorsolateral striatum did not affect striatal dopamine, glutamate and GABA release or nigral GABA release. Intrastriatal perfusion with the N-methyl-D-aspartate receptor antagonist dizocilpine maleate (10 microM), at a dose which by itself did not affect basal striatal or nigral neurotransmitter levels, prevented the effects of striatal perfusion with N-methyl-D-aspartate on both striatal and nigral neurotransmitter release. Intrastriatal dizocilpine maleate was also perfused concurrently with intranigral tetrodotoxin (10 microM) (see accompanying paper). Intrastriatal perfusion with dizocilpine maleate prevented the tetrodotoxin induced rise in both striatal and nigral GABA levels and profoundly reduced the tetrodotoxin-induced contralateral turning. In addition, intrastriatal dizocilpine maleate delayed the increase in striatal glutamate release evoked by intranigral tetrodotoxin without affecting the associated decrease in striatal dopamine release. The present study demonstrates that N-methyl-D-aspartate receptors in the dorsolateral striatum regulate GABA release in the substantia nigra pars reticulata of the awake rat and provides evidence that this regulation plays a key role in motor function. PMID- 8730709 TI - Subchronic administration of clozapine, but not haloperidol or metoclopramide, decreases dopamine D2 receptor messenger RNA levels in the nucleus accumbens and caudate-putamen in rats. AB - The effects of unique profile antipsychotic drugs on dopamine D2 receptors and D2 receptor messenger RNA were assessed following subchronic administration in rats. Male, Sprague-Dawley rats were administered oral haloperidol, clozapine, metoclopramide or no drug for three weeks via their drinking water. Tissue from the medial nucleus accumbens and dorsolateral caudate-putamen was dissected and analyzed by Northern blot analysis for levels of dopamine D2 receptor messenger RNA and binding assays conducted with [3H]spiperone for dopamine D2 receptors. Haloperidol and metoclopramide, but not clozapine, significantly increased [3H]spiperone in the caudate-putamen, but not the nucleus accumbens. Clozapine significantly decreased D2 messenger RNA levels in the caudate-putamen and the nucleus accumbens, while metoclopramide and haloperidol had no significant effect in either brain region. The finding of decreased D2 receptor messenger RNA levels produced by subchronic clozapine may account for the lack of striatal D2 receptor up-regulation, which was robustly observed after subchronic haloperidol and metoclopramide. Furthermore, since haloperidol and metoclopramide have a high liability for motor side effects, the current results relate favorably to the low motor side effect profile of clozapine. PMID- 8730710 TI - Increased subthalamic neuronal activity after nigral dopaminergic lesion independent of disinhibition via the globus pallidus. AB - Electrophysiological records of unit activity were used to compare the effects of excitotoxic pallidal lesions and 6-hydroxydopamine-induced damage to the midbrain dopaminergic neurons on the discharge rates and patterns of the subthalamic neurons. Removal of the pallidal input induced a slight, but statistically significant, increase (19.5%) in the discharge rate and no change in the firing pattern when compared to control animals. The rats with a dopaminergic lesion showed greater increase (105.7%) while the firing pattern activity of the subthalamic neurons became more irregular, with burst. These results indicate that the increased activity of the subthalamic neurons following a midbrain dopaminergic lesion cannot be due solely to inhibition-disinhibition involving the striato-pallido-subthalamic pathway and induced by the striatal dopaminergic depletion. PMID- 8730711 TI - A role for the subthalamic nucleus in 5-HT2C-induced oral dyskinesia. AB - The 5-hydroxytryptamine2C serotonin receptor is broadly distributed in brain, however, its functional role is unknown. Peripheral administration of drugs acting at the 5-hydroxytryptamine2C receptor induces abnormal oral dyskinesias, hyperkinetic motor disorders that often result from dysfunction of the basal ganglia. The subthalamic nucleus, a brain region anatomically and functionally related to the basal ganglia, has been implicated in oral dyskinesia. The subthalamic nucleus contains messenger RNA encoding 5-hydroxytryptamine2C receptors, suggesting its potential role in 5-hydroxytryptamine2C-mediated oral dyskinesia. Both systemic administration and local unilateral infusion of the 5 hydroxytryptamine2C/1B agonist, 1-(m-chlorophenyl)piperazine into the subthalamic nucleus increased orofacial movements. Oral movements following subthalamic infusion of 1-(m-chlorophenyl)piperazine were blocked by systemic administration of the 5-hydroxytryptamine2C/2A antagonists mianserin, ketanserin and mesulergine but were not altered by systemic pretreatment with either the 5 hydroxytryptamine1A/2A and dopamine antagonist spiperone or the 5 hydroxytryptamine1A/1B antagonist pindolol. Co-infusion of mesulergine with 1-(m chlorophenyl)piperazine into the subthalamic nucleus blocked 1-(m chlorophenyl)piperazine-stimulated oral movements. Oral bouts following systemically administered 1-(m-chlorophenyl)piperazine were markedly reduced following bilateral subthalamic infusion of either mesulergine or the selective 5 hydroxytryptamine2C antagonist SDZ SER 082. The findings indicate that stimulating 5-hydroxytryptamine2C receptors in the subthalamic nucleus elicits orofacial dyskinesia in the rat. These data are novel in providing a behavioral model for central 5-hydroxytryptamine2C receptor stimulation attributed to a specific anatomical location, and suggest that antagonists at the 5 hydroxytryptamine2C receptor could be useful in treating hyperkinetic motor disorders. PMID- 8730712 TI - P-chloroamphetamine induces c-fos in rat brain: a study of serotonin2A/2C receptor function. AB - Recent studies suggest that a class of substituted amphetamines, which includes p chloroamphetamine, causes an acute release of serotonin (5-hydroxytryptamine) and appears to act preferentially on axons arising from the dorsal raphe nucleus. The postsynaptic targets of these axons are not well characterized, but they have been localized in close proximity to the distribution of serotonin2A receptor binding sites. In the present study, c-fos immunocytochemistry has been used to investigate this anatomical relationship further. Administration of p chloroamphetamine or the serotonin2A/2C receptor agonist 1-(2,5-dimethoxy-4 iodophenyl)-2-aminopropane to rats resulted in similar patterns of Fos-like immunoreactivity in some, but not all, forebrain areas. Areas which expressed Fos following either treatment included cerebral cortex, claustrum, amygdala and nucleus accumbens. A particularly close match was seen in layer Va of the somatosensory cortex. No specificity of p-chloroamphetamine for dorsal raphe nucleus-innervated areas was noted. Prior treatment of animals with p chloroamphetamine two weeks before a second challenge with the same drug, or with the serotonin2A/2C receptor antagonist ritanserin 30 min before p chloroamphetamine challenge, resulted in an attenuation of p-chloroamphetamine induced Fos-like immunoreactivity in the olfactory tubercle, the islands of Calleja and the caudate-putamen. The reduction was most noticeable in layer Va of the somatosensory cortex. The results of this study indicate that a close anatomical correlation may exist between the fine serotonin axon terminals that show vulnerability to the neurotoxic effects of p-chloroamphetamine and serotonin2A receptors in some brain regions. This association may prove to be important in explaining the actions of certain psychotropic drugs, for example in the control of affective states. PMID- 8730713 TI - Burst stimulation of the medial forebrain bundle selectively increase Fos-like immunoreactivity in the limbic forebrain of the rat. AB - The present study was designed to evaluate the postsynaptic functional consequences of different presynaptic activity patterns in midbrain dopamine systems using electrical stimulation of the rat medial forebrain bundle and subsequent determination of c-fos expression, used as a marker for neuronal activation, in dopamine target areas, by means of Fos immunohistochemistry. Nerve terminal dopamine release evoked by electrical stimulation of the medial forebrain bundle was monitored in the same animals using in vivo voltammetry. A 5 Hz stimulation consisting of 60 trains of five pulses and lasting 1 min was applied to the medial forebrain bundle. This stimulation was repeated 15 times every 3 min. Its pattern was defined by the interpulse interval which was either 70 ms or 200 ms for burst or regularly spaced stimulation, respectively. Our results show that burst stimulation of the medial forebrain bundle, which increase release of dopamine in target areas, increases the basal Fos-like immunoreactivity in the stimulated hemisphere, while regular stimulation does not affect expression of this protein. Moreover, the increase in Fos-like immunoreactivity induced by burst stimulation is restricted to limbic related structures, i.e. nucleus accumbens shell and intermediate aspect of the lateral septum, and the major island of Calleja, but is not observed in motor related structures (nucleus accumbens core and striatum). Pretreatment with the D1 dopamine receptor antagonist, SCH23390 (0.1 mg/kg, i.p.), blocked the increase in Fos-like immunoreactivity induced by burst stimulation of the medial forebrain bundle, suggesting a role for these receptors in the observed effects. Pretreatment with the 5-hydroxytryptamine2A/2C receptor antagonist ritanserin (0.4 mg/kg, i.p.) did not affect the increase in Fos-like immunoreactivity induced by burst stimulation in the nucleus accumbens shell or in the lateral septum, although it blocked the stimulated enhancement of Fos-like immunoreactivity in the major island of Calleja. The present data indicate that, rather than the absolute mean discharge rate of midbrain dopamine neurons, the temporal organization of the action potentials they generate conveys information to their target areas. PMID- 8730714 TI - An experimental model for the non-invasive trans-synaptic induction of nitric oxide synthase in Purkinje cells of the rat cerebellum. AB - Nitric oxide synthase and its product nitric oxide are believed to be significantly involved in neuronal degeneration associated with various excitotoxic events and in some acute and chronic neuropathological disease states. The synchronous activation of inferior olive neurons by the drug harmaline is known to produce tremorogenic activity in rats via increased stimulation of the olivocerebellar system. Following subcutaneous or intraperitoneal injection (15, 30 or 60 mg/kg) of harmaline and survival intervals of three days or more, NADPH-diaphorase histochemistry and nitric oxide synthase immunocytochemistry revealed nitric oxide synthase induction in Purkinje cells of the cerebellar vermis and paravermis, but not the lateral hemispheres. Double labeling with NADPH-diaphorase and calbindin revealed that induced Purkinje cells contain calbindin and that induced Purkinje cells are not necessarily associated with regions of Purkinje cell degeneration. The induction of nitric oxide synthase in a subpopulation of Purkinje cells may reflect a protective response of selected Purkinje cells to overstimulation of specific olivocerebellar projections. This is the first experimental model for the non invasive induction of nitric oxide synthase in central nervous system neurons and as such may provide a useful tool for studying the induction of nitric oxide synthase associated with excitotoxic events. PMID- 8730715 TI - Site-specific regulation of Alzheimer-like tau phosphorylation in living neurons. AB - The microtubule-associated protein tau is more highly phosphorylated at certain residues in developing brain and in Alzheimer's disease paired helical filaments than in adult brain. We examined the regulation of tau phosphorylation at some of these sites in rat brain using the phosphorylation state-dependent anti-tau antibodies AT8, Tau1, and PHF1. The AT8 and PHF1 antibodies bind to phosphorylated tau, while Tau1 binds to unphosphorylated tau. Levels of tau reactive for AT8 were high only during the first postnatal week, with levels in adult declining to approximately 5% of the levels in neonates. In neonatal forebrain slices, tau became rapidly dephosphorylated at the AT8 and Tau1 sites during incubation at 34 degrees C, but was incompletely dephosphorylated at the PHF1 site. Dephosphorylation at AT8 sites, but not at Tau1 or PHF1 sites, was completely inhibited by 1 microM okadaic acid. Hence the regulation of tau phosphorylation by okadaic acid-sensitive phosphatase(s) was site-specific. Addition of 1 microM okadaic acid after dephosphorylation at the AT8 locus yielded a partial recovery of AT8 immunoreactivity, and incubation with basic fibroblast growth factor increased phosphorylation at the AT8 site in a dose dependent manner. These results indicate that endogenously active and basic fibroblast growth factor stimulated tau kinases directed toward an Alzheimer's disease-related site were present in the slices. In adult brain slices, the small pool of AT8-reactive tau was remarkably insensitive to dephosphorylation during incubation, and okadaic acid treatment induced only small increases in AT8 immunoreactivity. These results suggest that tau phosphorylation in adult brain is less dynamic than in neonatal brain. These findings indicate that neonatal tau is not only phosphorylated more highly than adult tau, but also more dynamically regulated by protein phosphatases and protein kinases than adult tau. The inability of okadaic acid to induce large increases in tau phosphorylation in adult rat brain slices suggests that a loss of protein phosphatase activity alone would not be sufficient to produce the hyperphosphorylation observed in Alzheimer's disease paired helical filaments. Stimulation of kinase activity by basic fibroblast growth factor is likely to modulate tau function during development, and may contribute to the genesis of hyperphosphorylated tau in Alzheimer's disease. PMID- 8730716 TI - Ascending and descending projections from the rostral nucleus of the solitary tract originate from separate neuronal populations. AB - Anterograde studies have shown that neurons within the rostral (gustatory) nucleus of the solitary tract project to the parabrachial nucleus, as well as to sites within the medulla including the reticular formation and caudal nucleus of the solitary tract. In order to determine the degree to which the same neurons contribute to both projections, injections of retrograde tracers were made simultaneously into both the parabrachial nuclei and medullary reticular formation of the rat. Only a small proportion of neurons were double labeled. Consistent with studies in hamster, labeled neurons projecting to the parabrachial nuclei in rat consisted of both stellate and elongate neurons, concentrated within the central subdivision of the rostral nucleus of the solitary tract. Injections into the medullary reticular formation also labeled both stellate and elongate neurons but these were concentrated in the ventral subdivision of the nucleus. The results of the present study demonstrate that different populations of neurons in the nucleus of the solitary tract contribute to ascending and descending pathways. This suggest a possible functional specialization within the nucleus of the solitary tract for those neurons whose output eventually reaches the forebrain compared to those neurons with local connections. PMID- 8730717 TI - The vibration pattern of the hearing organ in the waltzing guinea-pig measured using laser heterodyne interferometry. AB - The mechanical tuning characteristics of the hearing organ were measured in response to sound stimulation using laser heterodyne interferometry in in vitro preparations of temporal bones from waltzing guinea-pigs expressing different degrees of hearing organ and sensory cell degeneration. Measurements were made at various stages of structural changes allowing us to correlate structure and mechanical function. It was found that the characteristic frequency of the response at a given location in the cochlea occurred at lower frequencies than what is normally seen and that the sharpness of the mechanical tuning was considerably reduced when sensory hair cells were absent and the hearing organ structurally altered. However, even when extensive hair cell degeneration was evident a residual mechanical tuning was present. These results further support the concept that the sensory hair cells plays a key role in determining normal auditory tuning characteristics. It is suggested that the basilar membrane mechanics gives rise to a broadly tuned mechanical response on which a sharper tuning mechanism, originating from the hair cells, is superimposed. PMID- 8730718 TI - Methamphetamine modifies the photic entraining responses in the rodent suprachiasmatic nucleus via serotonin release. AB - We examined whether methamphetamine modifies the photic entraining responses in the rat suprachiasmatic nucleus. Optic nerve stimulation increased vasoactive intestinal polypeptide release from rat suprachiasmatic nucleus slices, and methamphetamine inhibited this increase in a concentration-dependent manner. Optic nerve stimulation has been reported to evoke field potentials in rat suprachiasmatic nucleus slices. Methamphetamine attenuated this field potential, and maximal inhibition (75.5%) was achieved at a concentration of 100 microM. Systemic administration of methamphetamine (1-5 mg/kg) inhibited light (300 lux, 1h)-induced Fos expression in the suprachiasmatic nucleus; methamphetamine at a dose of 5 mg/kg, i.p. caused 40% inhibition of light-induced Fos expression. We examined whether the inhibitory effect of methamphetamine on photic entraining responses mediates serotonin release from the suprachiasmatic nucleus. High performance liquid chromatographic analysis revealed that methamphetamine application increased serotonin release from rat suprachiasmatic nucleus slices in a concentration-dependent manner, but did not affect noradrenaline release. In addition, reduction of serotonin content attenuated the effect of methamphetamine on field potential induced by optic nerve stimulation in vitro and also light induced phase advances of wheel running activity rhythm in vivo. The present results support the idea that methamphetamine produces an inhibitory effect on photic entrainment in the suprachiasmatic nucleus via serotonin release. PMID- 8730719 TI - Immunocytochemical evidence for the presence of vasopressin in intermediate sized neurosecretory granules of solitary neurohypophyseal terminals in the homozygous Brattleboro rat. AB - A single base deletion (delta G) in the vasopressin gene is the cause of diabetes insipidus in the homozygous Brattleboro rat (di/di). The resulting frameshift leads to the expression of an aberrant vasopressin precursor which is unable to enter the secretory pathway, thereby preventing vasopressin biosynthesis. In a small number of solitary magnocellular hypothalamic neurons within the supraoptic and paraventricular nuclei, the reading frame is restored by a dinucleotide (delta GA) frameshift mutation, at two separate GAGAG motifs downstream of the original G-deletion. This results in two + 1 di-vasopressin precursors that are still partially mutated within the neurophysin region. The present study provides immunocytochemical evidence which demonstrates that, within magnocellular solitary neurons of the supraoptic and paraventricular nuclei of the di/di rat, the + 1 di-vasopressin precursors can enter the secretory pathway followed by their enzymatic processing into vasopressin during axonal transport to the neural lobe. However, the cellular characteristics of biosynthesis are different from those of wild-type rats. Immunoelectron microscopical localization of vasopressin gene products in the neural lobe of did/di rats revealed their presence in neurosecretory granules, the diameter of which is intermediate (116 nm) between those of the neurosecretory granules in the di/di (80-100 nm) and wild-type (160 nm) rats. PMID- 8730720 TI - Spinal cord mechanisms of opioid tolerance and dependence: Fos-like immunoreactivity increases in subpopulations of spinal cord neurons during withdrawal [corrected]. AB - Tolerance to the analgesic effects of morphine results in part from the development of a compensatory response in neurons that express the opioid receptor or of neural circuits in which those neurons participate. According to this formulation, withdrawal of morphine results in an overshoot of several neuronal properties because of the unopposed action of the compensatory response system. To identify the population of spinal cord neurons that underlies this state, we monitored expression of Fos-like immunoreactivity, after naltrexone precipitated abstinence in normal and morphine-tolerant rats. After daily (five days) implantation of morphine or placebo pellets, the rats received an injection of saline or naltrexone and behavior was monitored for 1 h. The rats were then killed, their spinal cords removed and 50-microns transverse sections of the lumbar cord were immunostained with a rabbit polyclonal antiserum directed against Fos. Naltrexone injection in the placebo group did not increase spinal cord Fos expression. Naltrexone-precipitated abstinence resulted in an increase in Fos expression at all levels of the spinal cord; the greatest increase and densest staining was in laminae I through VI. Importantly, when withdrawal was precipitated in anesthetized rats, we recorded a significant reduction in Fos expression, particularly in laminae III through VI, but there was persistent expression in the superficial dorsal horn, particularly in lamina I. These results suggest that spinal cord nociresponsive neurons are sensitized during the development of tolerance. This sensitization is unmasked by the administration of naltrexone and is manifested by fos induction in laminae I/II in awake or anesthetized withdrawing animals. The underlying mechanisms of tolerance development may be similar to those that underlie injury-induced central sensitization and hyperalgesia. PMID- 8730721 TI - Differential distribution of Fos-like immunoreactivity in the spinal trigeminal nucleus after noxious and innocuous thermal and chemical stimulation of rat cornea. AB - Corneal afferent nerves project to two spatially distinct sites within the spinal trigeminal nucleus: the subnucleus interpolaris/caudalis transition and the subnucleus caudalis/upper cervical spinal cord transition. The role of these two regions in processing corneal input is uncertain. To determine if neurons in these regions encode different features of an applied corneal stimulus, immunoreactivity for the immediate early gene protein product, Fos, was quantified in barbiturate-anesthetized rats. Intensity was varied across thermal (thermal probe 5, 35, 42, 52 degrees C; radiant heat of approximately 45 degrees C) stimuli and compared with that seen after mustard oil (5 microliters, 20%) or mineral oil application. All stimuli increased the number of Fos-positive neurons located at the ventrolateral pole of the subnucleus interpolaris/caudalis transition compared with unstimulated controls. By contrast, only 52 degrees C thermal probe and mustard oil produced an additional peak of Fos-positive neurons within the superficial laminae at the subnucleus caudalis/cervical cord transition. Further, the magnitudes of the bimodal peaks of Fos produced by 52 degrees C thermal probe and mustard oil stimuli were different quantitatively. Mustard oil caused a greater Fos response at the subnucleus interpolaris/caudalis transition than 52 degrees C thermal probe stimulation, whereas the opposite was true at the subnucleus caudalis/cervical cord transition. Double-labeling revealed that Fos immunoreactive neurons within the spinal trigeminal nucleus were restricted to regions densely labeled for calcitonin gene-related peptide. These results indicate that select features of corneal stimuli such as modality are encoded differently by neurons in the trigeminal subnucleus interpolaris/caudalis transition compared with those located in the subnucleus caudalis/cervical cord transition. It is likely that neurons in these two brainstem regions subserve different aspects of corneal sensation. PMID- 8730722 TI - Neurokinin-1 receptors on lumbar spinothalamic neurons in the rat. AB - In order to determine whether spinothalamic neurons in the lumbar spinal cord of the rat process neurokinin-1 (substance P) receptors, we injected cholera toxin B subunit into the thalamus and carried out dual-labelling immunocytochemistry to search for neurons that were immunoreactive with antibodies to cholera toxin and neurokinin-1 receptor. We examined 356 spinothalamic neurons in transverse sections and found that 35% of these were neurokinin-1 receptor-immunoreactive. Double-labelled cells made up the majority of the spinothalamic population in lamina I and the lateral spinal nucleus, and were also present in laminae III-V and the area around the central canal. On the side contralateral to the injection site, 77% of spinothalamic neurons in lamina I also showed neurokinin-1 receptor immunoreactivity, while 33% of those in laminae III-V and 14% of the ventromedial group possessed the receptor. Several of the double-labelled neurons with cell bodies in laminae III and IV had dendrites which could be followed dorsally into the superficial dorsal horn. These results indicate that substance P released from nociceptive primary afferents into the superficial dorsal horn is likely to act on spinothalamic tract neurons in lamina I, and also on those with cells bodies in laminae III-IV and long dorsal dendrites. PMID- 8730723 TI - Target-dependent plasticity of galanin and vasoactive intestinal peptide in the rat superior cervical ganglion after nerve lesion and re-innervation. AB - The expression of the neuropeptides galanin and vasoactive intestinal peptide (VIP) is increased in subpopulations of sympathetic neurons after axotomy of the rat superior cervical ganglion. We investigated whether postganglionic neurons innervating different targets show a prevalence for any of the two peptides in response to carotid nerve lesion. Before the respective postganglionic axons were crushed close to the ganglion, postganglionic neurons projecting either to the iris (through the internal carotid nerve) or to the submandibular gland (through both carotid branches) were identified by the retrograde tracer Fast Blue. Galanin and VIP immunoreactivities were demonstrated two and 30 days after crush and after successful regeneration of the lesioned neurons (60 days). In control ganglia, both peptides were detected in a few gland- but not in iris-projecting neurons. However, two days after crush of the respective carotid nerves, 14% of neurons within the iris and 46% within the gland population were immunoreactive for galanin. The percentage of neurons immunoreactive for VIP was significantly lower in both populations: only 3.5% of neurons projecting to the iris and 23% of the gland-projecting neuron population exhibited this peptide. After 30 days, the percentage of galanin- and VIP-positive neurons projecting to the submandibular gland was reduced (24% and 5.7%, respectively), whereas the proportion of galanin immunoreactive neurons further increased within the iris population (55%), indicating that some neurons express galanin at later stages after the lesion. At 60 days after the crush, the percentage of galanin- or VIP-immunoreactive neurons had decreased to control levels within those neuron populations that re innervated the iris or submandibular gland, although the total number of neurons exhibiting galanin or VIP was still increased within the ganglion, suggesting that re-establishment of target contact may play a role in down-regulation of both peptides. PMID- 8730724 TI - Differential activation of microglia and astrocytes following trimethyl tin induced neurodegeneration. AB - We have investigated the response of astrocytes and microglia to trimethyl tin intoxication in the septum, hippocampus, olfactory bulb, and pyriform cortex of the rat. Microglia were studied qualitatively using lectin histochemistry, and astrocytes were examined both qualitatively with immunohistochemistry, and quantitatively using an immunoassay for glial fibrillary acidic protein. Our results show that activated microglia first appeared 2 days after trimethyl tin intoxication in the lateral septum and hippocampus. Four days after trimethyl tin intoxication, the same regions revealed a most intense microglial reaction characterized by microglial hypertrophy and the formation of phagocytic clusters. By day 7, microglial activation in the septum and hippocampus had lessened, suggesting that the cells were reverting to the resting phenotype. The microglial response in the pyriform cortex and olfactory bulb, while being later in onset than in the septum and hippocampus, showed a similar progression of microglial changes reaching maximal intensity 7 days after trimethyl tin intoxication. Significant increases in the expression of glial fibrillary acidic protein were observed in all regions examined and typically occurred after microglial activation was already underway. We conclude that microglial and astroglial reactions which occur in response to trimethyl tin-induced neuronal necrosis are separated in time, with microglial activation preceding astrogliosis. In addition, our study stresses the importance of microglia as an endogenous source of CNS macrophages, and illustrates the merit of histochemical analysis with microglial markers for the early delineation of neurotoxicant-induced brain damage. PMID- 8730725 TI - Early history of IBRO: the birth of organized neuroscience. AB - To celebrate the thirtieth anniversary of the founding of the International Brain Research Organization (IBRO), a symposium was organized for the III Congress of the World Federation of Neuroscientists in Montreal to present, on 7 August 1991, the recollections of some of the early participants in IBRO's history and to place it in the context of other significant world organizations. Edited and slightly condensed transcripts of the talks together with the discussions are presented. L. Marshall reviewed the antecedent conferences that led to the Moscow Colloquium in 1958 and the subsequent formation of IBRO with UNESCO support. The ambience of that period for scientific organization on an international level was discussed by W. A. Rosenblith who emphasized how IBRO goals fitted into the rising interest in brain and behavior research. The scientific careers of six past executive secretaries were briefly reviewed by P. Gloor, with special tribute to the first, Herbert Jasper. One of IBRO's major achievements, the world survey of resources and needs in brain research, was described first-hand by G. Krauthamer. To discuss IBRO's image, C. Blakemore invoked the travails of reorganization and recovery from a period of inactivity. Finally, placing IBRO in a national and international context, S. Cozzens presented the perspective of the historian of sociology on an upbeat note. PMID- 8730726 TI - Full sensitivity of P2X2 purinoceptor to ATP revealed by changing extracellular pH. AB - A full pharmacological characterization was carried out on a recombinant ATP gated ion channel (P2X2 purinoceptor) expressed in Xenopus oocytes. This slowly desensitizing neuronal P2X2 purinoceptor, activated by ATP (EC50 = 4.6 +/- 1 microM at pH 7.4; n = 4), showed the agonist potency order: ATP > or = 2-MeSATP = ATP gamma S > or = ATP alpha S > > Bz-ATP. The receptor affinity for ATP was enhanced 5-10 fold by acidifying the bathing solution (to pH 6.5) but was diminished 4-5 fold in an alkaline solution (pH 8.0). The maximum activity of P2X2 purinoceptors and the activity order of a series of nucleotides were unaltered by changing extracellular pH. Interestingly, ATP sensitivity at a recombinant P2Y1 purinoceptor remained unaltered with changing extracellular pH. These results indicate that acidotic conditions in the synaptic cleft could strengthen purinergic transmission at neuronal P2X2 purinoceptors. PMID- 8730727 TI - Functional evidence of atypical beta 3-adrenoceptors in the human colon using the beta 3-selective adrenoceptor antagonist, SR 59230A. AB - The role of beta 3-adrenoceptors in human colonic circular smooth muscle was assessed in vitro by use of the beta 3-selective antagonist SR 59230A. Isoprenaline, in the presence of the selective beta-adrenoceptor antagonists CGP 20712A (beta 1) and ICI 118551 (beta 2), both at 0.1 microM, concentration dependently relaxed the preparation (pEC50 = 5.22). This effect was potently and competitively antagonized by SR 59230A with a pA2 of 8.31, while its R,R enantiomer SR 59483A gave an apparent pKB of 6.21. Relaxation was likewise produced by CGP 12177A (pEC50 = 6.05), but not by BRL 37344. Although only one of these beta 3-selective agonists was effective, the remarkably high potency of SR 59230A as a stereospecific antagonist of non-beta 1 non-beta 2 relaxation of human colonic muscle by isoprenaline provides strong functional evidence of beta 3-adrenoceptors in that tissue. PMID- 8730728 TI - Cisapride-induced prolongation of cardiac action potential and early afterdepolarizations in rabbit Purkinje fibres. AB - Cisapride, a gastrointestinal prokinetic agent, has been associated with cases of Torsades de Pointes but its effects on the cardiac action potential have not been described. We investigated its electrophysiological effects on rabbit isolated Purkinje fibres. The results demonstrated that cisapride (0.01-10 microM) lengthened concentration-dependently the action potential duration without modifying other parameters and induced early after depolarizations and subsequent triggered activity. This typical class III antiarrhythmic effect, that showed "reverse" rate-dependence and was reduced by increasing external K concentration, can account for clinical arrhythmogenesis. PMID- 8730729 TI - Binding of the radioligand [3H]-SCH 58261, a new non-xanthine A2A adenosine receptor antagonist, to rat striatal membranes. AB - 1. The present study describes the binding to rat striatal A2A adenosine receptors of the new potent and selective antagonist radioligand, [3H]-5-amino-7 (2-phenylethyl)-2-(2-furyl)-pyrazolo[4,3-e]-1,2,4-triazol o [1,5-c] pyrimidine, [3H]-SCH 58261. 2. [3H]-SCH 58261 specific binding to rat striatal membranes ( > 90%) was saturable, reversible and dependent upon protein concentration. Saturation experiments revealed that [3H]-SCH 58261 labelled a single class of recognition sites with high affinity (Kd = 0.70 nM) and limited capacity (apparent Bmax = 971 fmol mg-1 of protein). The presence of 100 microM GTP in the incubation mixture did not modify [3H]-SCH 58261 binding parameters. 3. Competition experiments showed that [3H]-SCH 58261 binding is consistent with the labelling of A2A striatal receptors. Adenosine receptor agonists competed with the binding of 0.2 nM [3H]-SCH 58261 with the following order of potency: 2 hexynyl-5'-N-ethyl carboxamidoadenosine (2HE-NECA) > 5'-N ethylcarboxamidoadenosine (NECA) > 2-[4-(2-carboxyethyl)-phenethylamino]-5'-N ethylcarboxamidoadenosi ne (CGS 21680) > 2-phenylaminoadenosine (CV 1808) > R-N6 phenylisopropyladenosine (R-PIA) > N6-cyclohexyladenosine (CHA) = 2-chloro-N6 cyclopentyladenosine (CCPA) > S-N6-phenylisopropyladenosine (S-PIA). 4. Adenosine antagonists inhibited [3H]-SCH 58261 binding with the following order: 5-amino-9 chloro-2-(2-furyl)-[1,2,4]-triazolo[1,5-c] quinazoline (CGS 15943) > 5-amino-8-(4 fluorobenzyl)-2-(2-furyl)-pyrazolo [4,3-e]-1,2,4-triazolo [1,5-c] pyrimidine (8FB PTP) = SCH 58261 > xanthine amine congener (XAC) = (E,18%-Z,82%)7-methyl-8-(3,4 dimethoxystyryl)-1,3-dipropylxanthine (KF 17837S) > 8-cyclopentyl-1,3 dipropylxanthine (DPCPX) > or = 8-phenyltheophylline (8-PT). 5. The Ki values for adenosine antagonists were similar to those labelled with the A2A agonist [3H] CGS 21680. Affinities of agonists were generally lower. The A1-selective agonist, R-PIA, was found to be about 9 fold more potent than its stereoisomer, S-PIA, thus showing the stereoselectivity of [3H]-SCH 58261 binding. Except for 8-PT, the adenosine agonists and antagonists examined inhibited [3H]-SCH 58261 binding with Hill coefficients not significantly different from unity. 6. The present results indicate that [3H]-SCH 58261 is the first non-xanthine adenosine antagonist radioligand which directly labels A2A striatal receptors. High receptor affinity, good selectivity and very low non-specific binding make [3H] SCH 58261 an excellent probe for studying the A2A adenosine receptor subtype in mammalian brain. PMID- 8730730 TI - Anti-inflammatory, membrane-stabilizing interactions of salmeterol with human neutrophils in vitro. AB - 1. We have investigated the effects of salmeterol (0.3-50 microM) on several pro inflammatory activities of human neutrophils in vitro. 2. Oxidant production by FMLP- and calcium ionophore (A23187)-activated neutrophils was particularly sensitive to inhibition by low concentrations (0.3-3 microM) of salmeterol, while the responses of phorbol myristate acetate- and opsonised zymosan-stimulated cells were affected only by higher concentrations (3-50 microM) of the drug. At these concentrations salmeterol is not cytotoxic, nor does it act as a scavenger of superoxide. 3. These anti-oxidative interactions of salmeterol with neutrophils were insensitive to propranolol but could be eliminated by washing the cells, or by pretreatment with low concentrations (1-2 microM) of the pro oxidative, membrane-destabilizing phospholipids, lysophosphatidylcholine (LPC), platelet activating factor (PAF) and lysoPAF (LPAF). 4. At concentrations of 6.25 50 microM salmeterol interfered with several other activities of stimulated neutrophils, including intracellular calcium fluxes, phospholipase A2 activity and synthesis of PAF. 5. In an assay of membrane-stabilizing activity, salmeterol (25 and 50 microM) neutralized the haemolytic action of LPC, PAF and LPAF. 6. Of the other commonly used beta 2-adrenoceptor agonists, fenoterol, and formoterol, but not salbutamol, caused moderate inhibition of neutrophil oxidant generation by a superoxide-scavenging mechanism. However, unlike salmeterol, these agents possessed only weak membrane stabilizing properties. 7. We conclude that salmeterol antagonizes the pro-inflammatory, pro-oxidative activity of several bioactive lipids implicated in the pathogenesis of bronchial asthma, by a mechanism related to the membrane-stabilizing, rather than to the beta 2-agonist properties of this agent. PMID- 8730731 TI - Phorbol ester activation of chloride current in guinea-pig ventricular myocytes. AB - 1. Although earlier studies with phorbol esters indicate that protein kinase C (PKC) may be an important regulator of Cl- current (Icl) in cardiac cells, there is a need for additional quantitative data and investigation of conflicting findings. Our objectives were to measure the magnitude, time course, and concentration-dependence of Icl activated in guinea-pig ventricular myocytes by phorbol 12-myristate 13-acetate (PMA), evaluate its PKC dependence, and examine its modification by external and internal ions. 2. The whole-cell patch clamp technique was used to apply short depolarizing and hyperpolarizing pulses to myocytes superfused with Na(+)-, K(+)-, Ca(2+)-free solution (36 degrees C) and dialysed with Cs+ solution. Stimulation of membrane currents by PMA (threshold < or = 1nM, EC50 approximately equal to 14 nM, maximal 40% increase with > or = 100 nM) plateaued within 6-10 min. 3. PMA-activated current was time-independent, and suppressed by l mM 9-anthracenecarboxylic acid (9-AC). Its reversal potential (Erev) was sensitive to changes in the Cl- gradient, and outward rectification of the current-voltage (I-V) relationship was more pronounced with 30 mM than 140 mM Cl- dialysate. 4. The relative permeability of PMA-activated channels estimated from Erev measurements was I- > Cl- > > aspartate. Channel activation was independent of external Na+. 5. PMA failed to activate Icl in myocytes pretreated with 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H-7) or dialysed with pCa 10.5 solution. Lack of response to 4 alpha-phorbol 12, 13-didecanoate (alpha PDD) was a further indication of mediation by PKC. 6. Icl induced by 2 microM forskolin was far larger than that induced by PMA, suggesting that endogenous protein kinase A is a much stronger Cl- channel activator than endogenous PKC in these myocytes. 7. The macroscopic properties of PMA-induced Icl appear to be indistinguishable from those of PKA-activated Icl. We discount stimulation of PKA by PMA as an explanation, and conclude that endogenous PKC may activate PKA regulated Cl- channels in these myocytes. PMID- 8730733 TI - Paradoxical facilitation of acetylcholine release from parasympathetic nerves innervating guinea-pig trachea by isoprenaline. AB - 1. Previous studies have provided evidence that activation of beta-adrenoceptors on cholinergic nerve terminals can inhibit neurotransmission in the airways. However, in most cases, this conclusion has been based on indirect evidence obtained from mechanical experiments where changes in airways smooth muscle tone were measured. 2. We have assessed whether modulation of cholinergic neurotransmission by beta-adrenoceptor agonists is due to a pre- or post junctional action by investigating the effect of isoprenaline on contractile responses evoked by exogenous acetylcholine (ACh) and electrical field stimulation (EFS; 4 Hz, 40 V, 0.5 ms pulse width every 15 s), and on EFS-induced ACh release from cholinergic nerves innervating guinea-pig and human trachea. Furthermore, the subtype of beta-adrenoceptor which modulates neurotransmission and the potential role of cyclic AMP in this response were evaluated. 3. In guinea-pig trachea, isoprenaline (1 nM-1 microM) inhibited the contractile response evoked by exogenous ACh (1 microM) to a similar extent to that evoked by EFS (EC50 = 19.9 and 23 nM, respectively). 4. In epithelium-denuded guinea-pig strips treated with indomethacin (10 microM), isoprenaline significantly enhanced EFS-induced ACh release from cholinergic nerve terminals (by 36% at 0.3 microM). This effect was blocked by propranolol and ICI 118, 551 (each 0.1 microM). In contrast, isoprenaline failed to affect EFS-induced ACh release from parasympathetic nerves innervating human trachea. 5. To evaluate the role of cyclic AMP in the beta-adrenoceptor-induced facilitation of cholinergic neurotransmission, the effects of various cyclic AMP elevating drugs on ACh release were studied. Forskolin (10 microM) significantly augmented (by 17%) EFS induced ACh release, an effect which was not reproduced by 1,9-dideoxyforskolin (10 microM) which does not activate adenylyl cyclase. Similarly, the cyclic AMP analogue, 8-bromo-cyclic AMP (1 mM) and cholera toxin (1 microgram ml-1) facilitated ACh output by 22 and 47% respectively, whereas prostaglandin E2 (PGE2, 0.1 nM-1 microM) inhibited this response (by 67% at 1 microM). 6. Zardaverine (10 microM), a dual inhibitor of the phosphodiesterase (PDE)3 and PDE4 isoenzyme families, did not affect EFS-induced ACh release and failed to facilitate the actions of either isoprenaline or PGE2. Similarly, neither SK&F 94120 (10 microM) nor rolipram (10 microM), selective inhibitors of PDE3 and PDE4 respectively, significantly affected the release of ACh in response to EFS. 7. The result of this study suggests that isoprenaline facilitates cholinergic neurotransmission in guinea-pig, but not human, trachea by activation of pre junctional beta 2-adrenoceptors, an effect that may be mediated via activation of the cyclic AMP/cyclic AMP-dependent protein kinase cascade. Furthermore, the data presented herein illustrate the need to undertake direct measurements of neurotransmitter release when examining the effect of agents purported to act pre junctionally. PMID- 8730732 TI - Effects of theophylline and rolipram on antigen-induced airway responses in neonatally immunized rabbits. AB - 1. The effects of the xanthine, theophylline, a non-selective phosphodiesterase (PDE) inhibitor, and the phosphodiesterase type 4 (PDE 4) inhibitor, rolipram, were evaluated in a model of antigen-induced airway responses in the allergic rabbit. 2. Adult litter-matched NZW rabbits (2.5-3.9 kg), immunized within 24 h of birth with Alternaria tenuis antigen, were pretreated twice daily for 3 days with theophylline (3 mg kg-1, i.p) or rolipram (1 mg kg-1, i.p) prior to antigen challenge (Alternaria tenuis). For each drug-treated group, a parallel group of rabbits were pretreated with the appropriate vehicle. In all groups airway responsiveness to inhaled histamine and bronchoalveolar lavage (BAL) was performed 24 h before and after antigen-challenge. 3. Basal lung function in terms of resistance (RL, cmH2O 1(-1)s-1) and dynamic compliance (Cdyn, ml cmH2O 1) were unaltered by pretreatment with theophylline or rolipram compared to their respective vehicles 24 h prior to or post antigen challenge. 4. The acute bronchoconstriction induced by inhaled Alternaria tenuis aerosol was unaffected by pretreatment with theophylline or rolipram. 5. Airway hyperresponsiveness to inhaled histamine was indicated by reduced RL PC50 (2.4-3.5 fold) and Cdyn PC35 (2.5-2.6 fold) values 24 h after antigen challenge. Treatment with rolipram, but not theophylline, prevented the increase in responsiveness to inhaled histamine 24 h after antigen challenge. 6. Total cells per ml of BAL fluid increased 24 h after antigen challenge due to the recruitment of neutrophils and eosinophils. Antigen-induced increases in pulmonary neutrophils were unaffected; however, eosinophils were reduced 57.5% in theophylline and 82% in rolipram-treated rabbits. 7. Inhalation of Alternaria tenuis aerosol elicits an acute bronchoconstriction, followed 24 h later by an increased responsiveness to inhaled histamine and pulmonary neutrophil and eosinophil recruitment in the immunized rabbit. With the dosing regimes used, both rolipram and theophylline inhibited eosinophil recruitment, whilst only rolipram prevented the development of airway hyperresponsiveness. Neither agent inhibited the acute bronchoconstriction due to inhaled antigen. PMID- 8730734 TI - Inhibition of inducible nitric oxide synthase expression by novel nonsteroidal anti-inflammatory derivatives with gastrointestinal-sparing properties. AB - 1. The effects of novel nitric oxide-releasing nonsteroidal anti-inflammatory compounds (NO-NSAIDs) on induction of nitric oxide (NO) synthase by bacterial lipopolysaccharide (LPS) were examined in a murine cultured macrophage cell line, J774. 2. LPS-induced nitrite production was markedly attenuated by the nitroxybutylester derivatives of flurbiprofen (FNBE), aspirin, ketoprofen, naproxen, diclofenac and ketorolac, with each compound reducing accumulated nitrite levels by > 40% at the maximum concentrations (100 micrograms ml-1) used. 3. Further examination revealed that nitrite production was inhibited in a concentration-dependent (1-100 micrograms ml-1) manner by FNBE which at 100 micrograms ml-1 decreased LPS-stimulated levels by 63.3 +/- 8.6% (n = 7). The parent compound flurbiprofen was relatively ineffective over the same concentration-range, inhibiting nitrite accumulation by 24 +/- 0.9% (n = 3) at the maximum concentration used (100 micrograms ml-1). 4. FNBE reduced LPS-induced nitrite production when added to cells up to 4 h after LPS. Thereafter, FNBE caused very little or no reduction in nitrite levels. Furthermore NO-NSAIDs (100 micrograms ml-1) did not inhibit the metabolism of L-[3H]-arginine to citrulline by NO synthase isolated from LPS-activated macrophages. 5. Western blot analysis demonstrated that NO synthase expression was markedly attenuated following co incubation of J774 cell with LPS (1 microgram ml-1; 24 h) and FNBE (100 micrograms ml-1; 24 h). Thus taken together, these findings indicate that NO NSAIDs inhibit induction of NO synthase without directly affecting enzyme activity. 6. In conclusion our results indicate that NO-NSAIDs can inhibit the inducible L-arginine-NO pathway, and are capable of suppressing NO synthesis by inhibiting expression of NO synthase. The clinical implications of these findings remain to be established. PMID- 8730735 TI - Increase in action potential duration and inhibition of the delayed rectifier outward current IK by berberine in cat ventricular myocytes. AB - 1. In the present work, the effects of the antiarrhythmic drug, berberine, on action potential and ionic currents of cat ventricular myocytes were studied. 2. Berberine prolonged action potential duration in cat ventricular myocytes without altering other variables of the action potential. 3. The drug at concentrations of 0.3-30 microM blocked only the delayed rectifier (IK) current with an IC50 = 4.1 microM. Berberine produced a tonic block and a phasic block that was increased with the duration of the depolarizing pulse. The blocking effect on IK was use-dependent, but not frequency-dependent. 4. In cardiac preparations two delayed rectifier currents have been found: a rapid (IKr) current and a slow (IKs) current. In the present work it has been found that berberine at the concentrations used, selectively blocked IKr. 5. At concentrations higher than 10 microM it also decreased the transient outward (Ito1) current. The drug did not have effects on the inward rectifier (IK1) or the high threshold calcium current (Ica-L). 6. These results show that berberine is a specific potassium channel blocker. The increase in action potential duration induced by berberine can be explained mainly by its blocking effects on IK. PMID- 8730736 TI - The lack of a role for potassium channel opening in the action of relaxin in the rat isolated uterus; a comparison with levcromakalim and salbutamol. AB - 1. The effects of relaxin in vitro in the isolated uterus from the non-pregnant rat were compared with those of levcromakalim and salbutamol in tissue bath, 42K+ -efflux and electrophysiological studies, to determine whether relaxin exhibits the characteristics of an opener of KATP-channels. 2. In uterus exposed to oxytocin (0.2 nM), tetraethylammonium (TEA, 10 mM) and glibenclamide (10 microM) produced large rightward shifts of the log10 concentration-effect curve to levcromakalim (125 fold and 118 fold, respectively). TEA (10 mM) caused only small rightward shifts of the log10 concentration-effect curves to salbutamol and relaxin (5.2 fold and 7.5 fold respectively). Glibenclamide did not antagonize salbutamol or relaxin. 3. Levromakalim (0.2 and 2 microM) suppressed the spasm evoked by low ( < or = 40 mM) but not high ( > 40 mM) concentrations of KCl. Salbutamol (1.5 nM) inhibited the spasm evoked by low concentrations of KCl ( < or = 40 mM). Salbutamol (15 nM) and relaxin (3 and 30 nM) inhibited the spasm evoked by low and high concentrations of KCl (10-80 mM). 4. Relaxin (0.12 microM) did not produce an increase in 42K+-efflux from longitudinal segments of rat myometrium. Exposure of tissues to relaxin (0.12 microM), in the presence of diltiazem (1 microM) plus KCl (20 mM), resulted in a small increase in 42K+ efflux of short duration. 5. Electrophysiological recording showed that the phasic spasms of the uterus exposed to oxytocin (0.2 nM) were accompanied by bursts of spiking activity superimposed upon a plateau potential. Inhibition of the mechanical activity of the uterus by levcromakalim (2 and 10 microM), salbutamol (30 nM) or relaxin (0.18 microM) was accompanied by a reduction in the duration of the plateau potential and the number of spikes without membrane hyperpolarization. 6. Unlike levcromakalim, relaxin did not selectively inhibit the spasm evoked by low concentrations of KCl and was not markedly antagonized by TEA or glibenclamide. Under conditions where a cromakalim-induced increase of the 42K+-efflux rate has been demonstrated, relaxin had only a very small effect. In isolated uterus from the rat, in contrast to observations in vivo, relaxin did not exhibit the characteristics of an opener of KATP-channels suggesting that another mechanism accounts for its inhibitory action. PMID- 8730737 TI - Effects of the 5-HT2B receptor agonist, BW 723C86, on three rat models of anxiety. AB - 1. BW 723C86 (3 and 10 mg kg-1, s.c. 30 min pretest), a 5-HT2B receptor agonist, increased total interaction, but not locomotion in a rat social interaction test, a profile consistent with anxiolysis. 2. The effect of BW 723C86 in the social interaction test is likely to be 5-HT2B receptor-mediated as it was prevented by pretreatment with the 5-HT2C/2B receptor antagonist, SB 200646A, (1 and 2 mg kg 1, p.o., 1 h pretest) which did not affect basal levels of social interaction at the doses used. 3. An anxiolytic-like action was also observed in the rat Geller Seifter conflict test, where BW 723C86 (0.5-50 mg kg-1, s.c. 30 min pretest) modestly, but significantly increased punished, but not unpublished responding. 4. In a rat 5 min elevated x-maze test, BW 723C86 (1-10 mg kg-1, s.c.) had no significant effect. 5. The maximal anxiolytic-like effect of BW 723C86 approached that of the benzodiazepine anxiolytic, chloradiazepoxide (5 mg kg-1, s.c. 30 min pretest) in the social interaction test, but was markedly less in the Geller Siefter test. The effect of BW 723C86 was also clearly less than chlordiazepoxide in the elevated x-maze procedure where it had no significant effect. 6. In conclusion, BW 723C86 exerted an appreciable anxiolytic-like profile in a rat social interaction test, but had a weaker effect in the Geller-Siefter and was ineffective in the elevated x-maze test used. These effects are likely to be 5 HT2B receptor-mediated. PMID- 8730738 TI - The role of lipocortin-1 in dexamethasone-induced suppression of PGE2 and TNF alpha release from human peripheral blood mononuclear cells. AB - 1.Lipocortin-1 and its N-terminal derivatives exert potent inhibitory actions in various models of acute inflammation. The present study examined the ability of lipocortin (LC)-1 to suppress the release of the acute pro-inflammatory mediators, tumour necrosis factor (TNF alpha) and prostaglandin E2 (PGE2) from human peripheral blood mononuclear cells (PBMC) stimulated with lipopolysaccharide (LPS) or recombinant human interleukin-1 beta (rhIL-1 beta). 2. LPS (10 micrograms ml-1) stimulated release of TNF alpha and PGE2 from PBMC was significantly inhibited by (4 h) co-incubation of the cells with 10(-6) M dexamethasone (Dex), but not with 10(-9) M to 10(-7) M of a N-terminal fragment (amino acids 1-188) of recombinant human LC-1 (LC-1 fragment). However, Dex suppression of LPS-stimulated TNF alpha and PGE2 secretion from PBMC was reversed when polyclonal antibody to LC-1 fragment (1:10,000 dilution) was included in the medium. rhIL-1 beta (5 x 10(-8) M)-stimulated release of TNF alpha and PGE2 from PBMC (after 18 h) was abolished by co-incubation of the cells with 10(-7) M LC-1 fragment. 3. After incubation with Dex (4 h), cellular proteins from PBMC were immunoblotted using anti-LC-1 fragment antibody (which showed to cross-reactivity with human annexins 2 to 6). Dex caused no increase in immunoreactive (ir)LC-1 content of PBMC, although there was a three fold increase in the amount of a lower mass species with LC-1-like immunoreactivity. This was accompanied by the appearance of irLC-1 in the extracellular medium. 4. The results of the present study implicate endogenous LC-1 in glucocorticoid suppression of TNF alpha and PGE2 release from human PBMC and suggest an extracellular site of action for LC 1. LC-1 may also inhibit rhIL-1 beta-stimulated TNF alpha and PGE2 secretion from PBMC. PMID- 8730739 TI - Pharmacological antagonism of the actions of group II and III mGluR agonists in the lateral perforant path of rat hippocampal slices. AB - 1. An understanding of the physiological and pathological roles of metabotropic glutamate receptors (mGluRs) is currently hampered by the lack of selective antagonists. Standard extracellular recording techniques were used to investigate the activity of recently reported mGluR antagonists on agonist-induced depressions of synaptic transmission in the lateral perforant path of hippocampal slices obtained from 12-16 day-old rats. 2. The group III specific mGluR agonist, (S)-2-amino-4-phosphonobutanoate (L-AP4) depressed basal synaptic transmission in a reversible and dose-dependent manner. The mean (+/-s.e. mean) depression obtained with 100 microM L-AP4 (the maximum concentration tested) was 74 +/- 3% and the IC50 value was 3 +/- 1 microM (n = 5). 3. The selective group II mGluR agonists, (1S,3S)-1-aminocyclopentane-1, 3-dicarboxylate ((1S,3s)-ACPD) and (2S, 1'R, 2'R, 3'R)-2-(2',3'-dicarboxycyclopropyl)glycine (DCG-IV) also depressed basal synaptic transmission in a reversible and dose-dependent manner. The mean depression obtained with 200 microM (1S,3S)-ACPD was 83 +/- 8% and the IC50 value was 12 +/- 3 microM (n = 5). The mean depression obtained with 1 microM DCG-IV was 73 +/- 7% and the IC50 value was 88 +/- 15 nM (n = 4). 4. Synaptic depressions induced by the actions of 20 microM (1S,3S)-ACPD and 10 microM L-AP4 were antagonized by the mGluR antagonists (+)-alpha-methyl-4-carboxyphenylglycine ((+)-MCPG), (S)-2-methyl-2-amino-4-phosphonobutanoate (MAP4), (2S,1'S,2'S)-2 methyl-2(2'-carboxycyclopropyl)glycine (MCCG), (RS)-alpha-methyl-4 tetrazolylphenylglycine (MTPG), (RS)-alpha-methyl-4-sulphonophenylglycine (MSPG) and (RS)-alpha-methyl-4-phosphonophenylglycine (MPPG) (all tested at 500 microM). 5. (+)-MCPG was a weak antagonist of both L-AP4 and (1S,3S)-ACPD-induced depressions. MCCG was selective towards (1S,3S)-ACPD, but analysis of its effects were complicated by apparent partial agonist activity. MAP4 showed good selectivity for L-AP4-induced effects. 6. The most effective antagonist tested against 10 microM L-AP4 was MPPG (mean reversal 90 +/- 3%; n = 4). In contrast, the most effective antagonist tested against 20 microM (1S,3S)-ACPD induced depressions was MTPG (mean reversal 64 +/- 4%; n = 4). Both antagonists produced parallel shifts in agonist dose-response curves. Schild analysis yielded estimated KD values of 11.7 microM and 27.5 microM, respectively. Neither antagonist had any effect on basal transmission or on depressions induced by the adenosine receptor agonist, 2-chloroadenosine (500 nM; n = 3). 7. We conclude that both group II and group III mGluRs can mediate synaptic depressions induced by mGluR agonists in the lateral perforant path. The mGlur antagonists MTPG, MPPG and MAP4 should be useful in determining the roles of group II and III mGluRs in the central nervous system. PMID- 8730740 TI - Mesenteric arterial function in the rat in pregnancy: role of sympathetic and sensory-motor perivascular nerves, endothelium, smooth muscle, nitric oxide and prostaglandins. AB - 1. The effects of pregnancy on mesenteric arterial function were examined in constantly perfused (5 ml min-1) mesenteric arterial beds isolated from 21-day pregnant rats. The function of sympathetic and sensory-motor perivascular nerves, endothelium and smooth muscle was examined. The role of nitric oxide and prostaglandins in vasoconstrictor function was tested by use of NG-nitro-L arginine methyl ester (L-NAME; 100 microM) and indomethacin (10 microM), respectively. 2. Electrical field stimulation (EFS; 4-32 Hz, 1 ms, 90V, 30s) at basal tone elicited frequency-dependent vasoconstriction which was markedly reduced in preparations from pregnant rats at all frequencies. Vasoconstrictor responses to vasopressin and endothelin were also reduced in pregnancy and there was a trend towards a reduction in maximal responses to noradrenaline (NA). In contrast, there was no difference in vasoconstrictor responses to ATP, 5 hydroxytryptamine (5-HT) or angiotension II. 3. L-NAME (100 microM) augmented responses to EFS, NA, ATP and vasopressin in control mesenteric arterial preparations. In contrast, L-NAME augmented responses only to EFS in pregnancy, having no significant effect on responses to NA, ATP and vasopressin. 4. Indomethacin (10 microM) attenuated responses to NA and vasopressin, but not to EFS, in controls and in pregnancy. Responses to ATP were attenuated by indomethacin in controls but not in pregnancy. 5. Mesenteric preparations from pregnant rats were resistant to having tone raised by continuous perfusion with methoxamine. Despite an approximately 10 fold greater concentration of methoxamine, there was a significantly smaller increase in tone in preparations from pregnant, 34.27 +/- 4.8 mmHg (n = 11) compared to control, 65.92 +/- 5.4 mmHg (n = 11), rats. EFS (4-12 Hz, 60 V, 0.1 ms, 30s) in the presence of guanethidine (5 microM) to block sympathetic neurotransmission elicited frequency dependent vasodilatation due to activation of sensory-motor nerves. Percentage relaxations were similar in preparations from pregnant and non-pregnant rats. 6. Dose-dependent endothelium-dependent vasodilatations to acetylcholine and ATP were similar in preparations from pregnant and non-pregnant rats. Endothelium independent vasodilatation to sodium nitroprusside and to calcitonin gene-related peptide were also similar between the two groups. 7. There was no significant difference in the basal perfusion pressure of mesenteric arterial beds from control (21.3 +/- 1.0 mmHg, n = 24) and pregnant (20.2 +/- 1.2 mmHg, n = 23) rats. However, a step-wise increase in perfusate flow from 5 to 10, 15, 20 and 24ml min-1 produced smaller increases in perfusion pressure in pregnancy compared to the controls. L-NAME (100 microM) or indomethacin (10 microM) had no significant effect on the relationship between flow and perfusion pressure. 8. The present results show that prejunctional changes are involved in blunted sympathetic vasoconstriction of rat mesenteric arteries in pregnancy. Non specific postjunctional changes are implicated in the reduced constrictor responses to applied methoxamine, vasopressin and endothelin, but not to ATP. In contrast, sensory-motor nerves and endothelium-dependent and -independent vasodilatation was unchanged. The decrease in receptor-mediated mesenteric arterial constrictor responsiveness in pregnancy does not appear to be due to acute modulation by NO or prostaglandins, but may involve changes in the distensibility of the bed and/or changes in wall thickness. PMID- 8730741 TI - Investigation of endogenous nitric oxide vascular function in the carotid artery of cholesterol-fed rabbits. AB - 1. The function of endogenous nitric oxide (NO) at the level of vascular smooth muscle, was assessed in a popular experimental model of accelerated atherosclerosis, the cholesterol-fed rabbit. 2. Endothelium-dependent vasorelaxation in response to acetylcholine (ACh, 1 microM) was significantly impaired in the carotid artery from rabbits maintained on a 1% (W/W) cholesterol diet for 8-10 weeks. Furthermore, the ability of an inhibitor of nitric oxide synthase (NOS), NG-nitro-L-arginine methyl ester (L-NAME, 1-300 microM), to enhance the contractile reactivity to a submaximal concentration of noradrenaline (NA, 3 microM), was significantly attenuated in hypercholesterolaemia. 3. A significant linear correlation between the maximal contractile effect of L-NAME (300 microM) and maximal vasorelaxation to ACh (1 microM) was determined in the carotid artery from control rabbits. In contrast, no such linear correlation was found in the carotid artery from hypercholesterolaemic rabbits. 4. We conclude that there are lesions both in agonist-stimulated, endogenous NO-dependent vasorelaxation and in the regulation of vasoconstrictor reactivity by endogenous NO in the hypercholesterolaemic rabbit carotid artery. Furthermore, the normal linear relationship between the contractile effect of L-NAME and vasorelaxation to ACh is lost after cholesterol-feeding. PMID- 8730742 TI - 5-HT4 receptor-mediated modulation of 5-HT release in the rat hippocampus in vivo. AB - 1. In the present study, the ability of the 5-hydroxytryptamine, receptor (5-HT4 receptor) to modulate the release of 5-HT in the hippocampus of freely-moving rats was investigated by the in vivo microdialysis technique. 2. The 5-HT4 receptor agonist, renzapride (1.0-100 microM, administered via the microdialysis probe) increased extracellular hippocampal levels of 5-HT in concentration dependent manner (approximately 200% maximal increase). The ability of renzapride (100 microM, administered via the microdialysis probe) to elevate extracellular levels of 5-HT remained in the presence of the selective 5-HT reuptake blocker, paroxetine (1.0 microM, administered via the microdialysis probe). Furthermore, another 5-HT4 receptor agonist 5-methoxytryptamine (5-MeOT; 10 microM, administered via the microdialysis probe, in the presence of the non-5-HT4 5-HT receptor antagonists pindolol (10 microM) and methysergide (10 microM)) maximally elevated extracellular levels of 5-HT by approximately 450% in the rat hippocampus. The elevation of extracellular 5-HT levels induced by either renzapride (100 microM) or 5-MeOT (10 microM) was completely prevented by combined administration of the selective 5-HT4 receptor antagonist, GR113808 (100 nM, administered via the microdialysis probe). GR113808 (100 nM, administered via the microdialysis probe) administered alone, however, reduced extracellular hippocampal 5-HT levels by some 60%. 3. Systemic administration of the 5-HT1A receptor agonist, 8-OH-DPAT (0.1 mg kg-1, s.c.) reduced extracellular levels of 5 HT in the rat hippocampus by approximately 40%. Prior administration of 8-OH-DPAT (0.1 mg kg-1, s.c.), with an associated reduction of extracellular hippocampal 5 HT levels by approximately 40-50%, however, failed to prevent a subsequent elevation of extracellular levels of 5-HT induced by renzapride (100 microM, administered via the microdialysis probe). 4. Systemic administration of the 5 HT4 receptor agonist, renzapride (0.25 and 1.0 mg kg-1, i.p.) increased extracellular levels of 5-HT in the hippocampus in a dose-dependent manner. The higher dose of renzapride increasing extracellular 5-HT levels by some 200%. The selective 5-HT4 receptor antagonist, GR125487D (1.0-100 micrograms kg-1, i.p.) caused a dose-dependent reduction in extracellular levels of 5-HT in the hippocampus (maximally approximately 80% reduction). Prior administration of GR125487D (10 micrograms kg-1, i.p.) prevented the elevation of extracellular levels of 5-HT induced by renzapride (1.0 mg kg-1, i.p.). 5. In conclusion, the present study provides evidence that activation of the 5-HT4 receptor facilitates 5-HT release in the rat hippocampus in vivo. PMID- 8730743 TI - Desipramine administration in the olfactory bulbectomized rat: changes in brain beta-adrenoceptor and 5-HT2A binding sites and their relationship to behaviour. AB - 1. The effects of repeated administration of the tricyclic antidepressant drug, desipramine (DMI), on behaviour (locomotor activity and rearing) and the number and affinity of brain beta-adrenoceptor and 5-HT2A receptor binding sites were examined in olfactory bulbectomized (OB) and sham-operated control rats. 2. Locomotor activity and rearing were increased in OB rats compared to sham operated controls. The effect of various doses of DMI (administered orally twice daily for 21 days) on these behavioural measures was examined. A dose of 7.5 mg kg-1 provided optimal reversal of hyperlocomotion and increased rearing in OB rats, without changing these measures in sham-operated controls. 3. The time course of DMI (7.5 mg kg-1) on behavioural and neurochemical measures was examined. locomotion and rearing in OB rats were not significantly altered after 7 days, were significantly attenuated after 14 days and were normalized after 21 days. 4. After 7 days of DMI administration the number of beta-adrenoceptors was lower in frontal and occipital cortex and hippocampus. This reduction was largely restricted to the beta 1-adrenoceptor subtype. Administration of DMI for 14 or 21 days did not further reduce the number of beta-adrenoceptors. The DMI induced reduction in beta-adrenoceptors did not differ in OB and sham-operated control rats. 5. DMI administration for up to 21 days produced a progressive reduction in the number of 5-HT2A receptors in frontal cortex, without significant alterations in occipital cortex. 6. The time course of the reduction in the number of 5-HT2A receptors was similar to that of the DMI-induced behavioural changes whereas that for the reduction in beta-adrenoceptors was clearly different. 7. The present results suggest that the action of DMI in this animal model is unlikely to be directly related to a reduction in beta-adrenoceptors but may be related to a reduction in frontal cortical 5-HT2A receptors. PMID- 8730744 TI - Effects of a partial agonist and a full antagonist acting at the glycine site of the NMDA receptor on inflammation-induced mechanical hyperalgesia in rats. AB - 1. NMDA receptor antagonists have previously been shown to have antinociceptive effects in behavioural experiments, but controversy remains as to the role of NMDA receptors in mechanical hyperalgesia. We have studied the effects on mechanical nociceptive thresholds in rats with carrageenin-induced paw inflammation of L-687,414, a low efficacy partial agonist which acts as a functional antagonist at the glycine modulatory site of the NMDA receptor and of L-701,324, a structurally novel, highly selective, full antagonist at this site. 2. Mechanical thresholds were measured for both hind paws 1 h before and 3 h after carrageenin or saline was injected into 1 hind paw. Dose-response curves were constructed for each test compound in separate experiments, with test compound or vehicle being given i.p. 1 h before the final test. 3. Both compounds produced selective dose-dependent and statistically significant reversal of mechanical hyperalgesia, with minimum effective doses of 100 mg kg-1 L-687,414 and 3 mg kg-1 L-701,324. Neither L-687,414 nor L-701,324 affected the response threshold of the contralateral non-inflamed paw over the dose-range producing reversal of carrageenin-induced hyperalgesia. Neither compound had any effect on the paw oedema produced by carrageenin injection. 4. These results show that both a full antagonist and a low efficacy partial agonist at the glycine modulatory site of the NMDA receptor complex reverse inflammation-induced mechanical hyperalgesia, thus supporting the argument that maximal activation of the glycine site is required for transmission via NMDA receptors, and showing that NMDA receptor-mediated actions are important in mechanical hyperalgesia induced by inflammation. PMID- 8730745 TI - Structure-activity relationships of new agonists and antagonists of different metabotropic glutamate receptor subtypes. AB - 1. We investigated the agonist and antagonist activities of 22 new phenylglycine and phenylalanine derivatives for metabotropic glutamate receptors (mGluRs) by examining their effects on the signal transduction of mGluR1, mGluR2 and mGluR6 subtypes expressed in Chinese hamster ovary cells. This analysis revealed several structural characteristics that govern receptor subtype specificity of the agonist and antagonist activities of phenylglycine derivatives. 2. Hydroxyphenylglycine derivatives possessed either an agonist activity on mGluR1/mGluR6 or an antagonist activity on mGluR1. 3. Carboxyphenylglycine derivatives showed an agonist activity on mGluR2 but an antagonist activity on mGluR1. 4. alpha-Methylation or alpha-ethylation of the carboxyphenylglycine derivatives converts the agonist property for mGluR2 to an antagonist property, thus producing antagonists at both mGluR1 and mGluR2. 5. Structurally corresponding phenylalanine derivatives showed little or no agonist or antagonist activity on any subtypes of the receptors. 6. This investigation demonstrates that the nature and positions of side chains and ring substituents incorporated into the phenylglycine structure are critical in determining the agonist and antagonist activities of members of this group of compounds on different subtypes of the mGluR family. 7. We also tested two alpha-methyl derivatives of mGluR agonists. (2S, 1'S, 2'S)-2-(2-Carboxycyclopropyl)glycine (L-CCG-I) is a potent agonist for mGluR2 but alpha-methylation of this compound changes its activity to that of an mGluR2-selective antagonist. In contrast, alpha-methylation of L-2 amino-4-phosphonobutyrate (L-AP4) results in retention of an agonist activity on mGluR6. Thus, alpha-methylation produces different effects, depending on the chemical structures of lead compounds and/or on the subtype of mGluR tested. PMID- 8730746 TI - Subtypes of losartan-sensitive angiotensin receptor in the rabbit pulmonary artery. AB - 1. The short rabbit pulmonary artery was denuded of endothelium and divided into three sections, the cardiac end (cardiac), middle and pulmonary end (pulmonary) sections, respectively. Des-Asp-angiotensin I attenuated the contractions of the cardiac and middle sections to transmural nerve stimulation but potentiated the contractions in the pulmonary section. 2. The actions of the nonapeptide were inhibited completely by 10(-6) M losartan; however, a similar concentration of PD123319 had no effect. Indomethacin (10(-6) M) also inhibited completely the attenuation in the cardiac and middle sections but had no effect on the potentiation seen in the pulmonary section. 3. The data suggest that the two differential responses of the pulmonary artery to des-Asp-angiotensin I are mediated by two separate subtypes of the losartan-sensitive angiotensin AT1 receptor. PMID- 8730747 TI - Actions of general anaesthetics on 5-HT3 receptors in N1E-115 neuroblastoma cells. AB - 1. NIE-115 mouse neuroblastoma cells were studied under voltage clamp in the whole-cell patch-clamp configuration. Peak currents induced by bath application of 5-hydroxytryptamine (5-HT) were inwardly rectifying, reversed at 0.4 +/- 0.2 mV (mean +/- s.e.mean), and were approximately half-inhibited (at 1 microM 5-HT) by 2 nM of the 5-HT3 selective antagonist MDL-72222 (3-tropanyl-3,5 dichlorobenzoate). 2. Peak inward currents activated by a low concentration of 5 HT at a holding potential of -50 mV were potentiated by volatile general anaesthetics. At their human minimum alveolar concentrations (MACs), the degree of potentiation increased in the order isoflurane < halothane < enflurane < methoxyflurane. Potentiation by methoxyflurane was independent of membrane potential in the range -70 mV to +40 mV. The reversal potential was the same in the presence and absence of methoxyflurane. 3. Methoxyflurane shifted the 5-HT dose-response curve to lower 5-HT concentrations, without significantly changing the Hill coefficient or maximum response. The EC50 concentration for 5-HT decreased from 1.86 +/- 0.02 microM to 1.07 +/- 0.11 microM (means +/- s.e.mean) due to the presence of 1 MAC (270 microM) methoxyflurane. 4. In contrast to the volatile anaesthetics, the barbiturate anaesthetic, thiopentone, inhibited the 5 HT3 receptor. Hill analysis of thiopentone dose-response data gave an average IC50 = 117 +/- 8 microM thiopentone and Hill coefficient = 1.6 +/- 0.2 (means +/- s.e.mean). These parameters were not significantly different for data obtained at 5-HT concentrations above and below the control EC50 concentration for 5-HT, consistent with non-competitive inhibition. 5. The n-alcohols occupied an intermediate position between the volatile and barbiturate anaesthetics. The lower alcohols (butanol and hexanol) potentiated 5-HT responses at low alcohol concentrations but inhibited them at high concentrations. In contrast, the higher alcohols (octanol, decanol, dodecanol, tridecanol, tetradecanol and pentadecanol) produced no potentiation, but only inhibition, at all alcohol concentrations. 6. Inhibition of the 5-HT3 receptor by the n-alcohols exhibited a cutoff in potency similar to those previously found for tadpoles, luciferase enzymes and a neuronal nicotinic acetylcholine receptor channel. PMID- 8730748 TI - The role of nephrectomy and proadifen in blood pressure homeostasis following an acute kinin-induced hypotension in normotensive rats. AB - 1. We have, in the present work, studied the importance of the kidneys and the renal hypotensive agent, medullipin, in modulating the blood pressure (BP) response to bradykinin, as well as their ability to influence the balance between the NO- and the adrenergic systems superimposed on a bradykinin-induced hypotension. 2. The rats were pretreated with the NO-synthase inhibitor, N omega nitro-L-arginine methyl esther (L-NAME) (0.3 g kg-1), proadifen (50 mg kg-1), an inhibitor of the medullipin-system, and nephrectomy (24 h) (Nx) alone, and L-NAME in combination with proadifen, Nx or phentolamine (2 mg kg-1). Subsequent injections of bradykinin (3, 6, 15, 30 micrograms kg-1) induced an acute hypotensive response. The fall in BP was dose-dependent in all groups (P < 0.01), except in the Nx/L-NAME group. No differences in the fall in BP were observed between the groups. 3. The duration of the hypotensive response was abbreviated after L-NAME-treatment (P < 0.05). Proadifen-treatment and Nx had no significant effect on the duration of the hypotension in control rats or in L-NAME-treated rats. Pretreatment with phentolamine prevented the L-NAME-induced rapid restoration of BP (P < 0.001). 4. In L-NAME-treated rats a transient hypertension followed the bradykinin-induced hypotensive response. This hypertensive response was not observed after Nx or proadifen-treatment alone, and addition of Nx or proadifen to L-NAME treatment did not alter the hypertensive response as compared to L-NAME alone. Phentolamine, however, abolished the L-NAME-induced hypertension (P < 0.05). 5. In conclusion, the present results do not support the involvement of the medullipin-system or other hypotensive systems localized in the kidneys, in modulating and counteracting the compensatory adrenergic response following an acute bradykinin-induced hypotension. A hampering modulating effect of the NO system on this compensatory adrenergic response was confirmed, indicating a close relationship between these two systems in BP homeostasis. PMID- 8730749 TI - The effects of saponin on the binding and functional properties of the human adenosine A1 receptor. AB - 1. Experiments with adenosine deaminase suggest that adenosine is present in membrane preparations from CHO cells bearing adenosine A1 receptors. 2. Pretreatment of the membranes (ca 0.6 mg protein ml-1) with the permeabilizing agent saponin (100 micrograms ml-1) or addition of saponin (10 micrograms ml-1) to the membranes (0.02-0.08 mg protein ml-1) in the assay, generates homogeneous low affinity agonist binding curves in the presence of GTP and an increased function, assessed by agonist stimulation of [35S]-GTP gamma S binding. The affinity constants for the binding of an agonist and an antagonist are not affected by this saponin treatment. Saponin facilitates the interaction of guanine nucleotides with receptor G-protein complexes, possibly by removing a permeability barrier to access of G-proteins by GTP. However, adenosine is still present in the binding assays after saponin treatment. 3. The agonist binding properties of the human A1 receptor have been characterized. In saponin pretreated membranes, 80-90% of the A1 receptors are capable of forming agonist receptor-G protein complexes in the absence of GTP. These complexes have a 300 600 fold higher affinity than uncoupled receptors for N6-cyclohexyladenosine. 4. A very slow component is observed in the association and dissociation kinetics of the agonist [3H]-N6-cyclohexyladenosine ([3H]-CHA) and in the association but not dissociation kinetics of the antagonist [3H]-8-cyclopentyl-1,3-dipropylxanthine ([3H]-DPCPX). The slow association component of [3H]-DPCPX is essentially absent when incubations are carried out in the presence of GTP. The slow dissociation component of [3H]-CHA binding is rapidly disrupted by GTP. 5. It is hypothesized that long-lasting adenosine-receptor-G protein complexes are present in the CHO membrane preparations. The existence of these complexes, resistant to the action of adenosine deaminase but sensitive to GTP, may rationalize the observed kinetics and the increase in 3H-antagonist binding produced by GTP which has been observed in essentially all studies of A1 receptors and has been ascribed previously to precoupling of A1 receptors to G-proteins in the absence of agonists. PMID- 8730750 TI - Regulation of tumour necrosis factor production by adrenal hormones in vivo: insights into the antiinflammatory activity of rolipram. AB - 1. The role of adrenal hormones in the regulation of the systemic and local production of tumour necrosis factor (TNF alpha) was examined in male Balb/c mice. 2. Intraperitoneal injection of 0.3 mg E. coli lipopolysaccharide (LPS, 0111:B4) led to high levels of circulating TNF alpha without stimulating TNF alpha production in the peritoneal cavity. Systemic production of TNF alpha in response to LPS was increased in adrenalectomized animals and in normal animals treated with the beta-adrenoceptor antagonist, propranolol. The glucocorticoid antagonist, RU 486, did not modify systemic TNF alpha production. These results indicate that systemic TNF alpha production is regulated by adrenaline but not by corticosterone. 3. When mice were primed with thioglycollate, TNF alpha was produced in the peritoneal cavity in response to low dose LPS (1 micrograms). The levels of TNF alpha in the peritoneal cavity were not enhanced by adrenalectomy or by treatment with either propranolol or RU 486, indicating local production of TNF alpha in the peritoneal cavity is not regulated by adrenaline or corticosterone. 4. The phosphodiesterase type IV (PDE-IV) inhibitor, rolipram, inhibited both the systemic production of TNF alpha in response to high dose endotoxin (ED50 = 1.3 mg kg-1) and the local production of TNF alpha in the peritoneal cavity in response to low dose endotoxin (ED50 = 9.1 mg kg-1). In adrenalectomized mice there was a slight reduction in the ability of rolipram to inhibit the systemic production of TNF alpha (ED50 = 3.3 mg kg-1) while the ability of rolipram to inhibit the local production of TNF alpha in the peritoneal cavity was virtually abolished (24% inhibition at 30 mg kg-1). The glucocorticoid antagonist, RU 486, also reduced the ability of rolipram to inhibit local TNF alpha production while propranolol was without effect. 5. Systemic treatment with rolipram increased the plasma concentrations of corticosterone in normal mice but not in adrenalectomized mice indicating that rolipram can cause adrenal stimulation in vivo. 6. In summary, these data indicate that systemic production of TNF alpha in response to high dose endotoxin is controlled differently from the local production of TNF alpha in response to low dose endotoxin. The systemic production of TNF alpha is regulated by catecholamines, but not by corticosterone, while the local production of TNF alpha in the peritoneal cavity is not regulated by basal levels of either catecholamines or corticosterone. 7. These data also show that the ability of rolipram to inhibit the local production of TNF alpha is dependent on the release of corticosterone from the adrenal glands. PMID- 8730751 TI - Unusual alpha-adrenoceptor subtype in canine saphenous vein: comparison to mesenteric vein. AB - 1. We investigated the nature of the adrenoceptors in the dog saphenous vein (DSV) and dog mesenteric vein (DMV) to determine the nature of the unexpected interactions of phenylephrine and methoxamine with rauwolscine in the DSV, i.e. the ability of the putative alpha 2-adrenoceptor antagonist to inhibit competitively contractions to these alpha 1-agonists. Radioligand binding studies were performed in parallel with contractility studies. 2. Functionally, in the DSV, phenylephrine and methoxamine-induced, contractions were antagonized by rauwolscine with Schild slopes of -0.52 and -0.46, respectively and apparent pA2 values of 8.5 and 9.2, respectively. Such antagonism was not observed in the DMV. In the DSV, prazosin competes for [3H]-rauwolscine binding sites with a high and a low affinity binding site (Ki of 1.49 +/- 0.65 and 94.7 +/- 51 microM, n = 6, respectively). 3. Pretreatment with 100 microM chloroethylclonidine (CEC) for 15 min abolished [3H]-prazosin binding in microsomes from both veins and reduced binding (Bmax) of [3H]-rauwolscine in microsomes by 55.1 +/- 0.8% (n = 3) in the DSV but did not affect the Bmax in the DMV. CEC pretreatment in the venular rings denuded of endothelium caused persistent contraction in the DSV but not in the DMV. In the DSV, CEC appeared to interact with a single [3H]-rauwolscine binding site. In both the DSV and the DMV, CEC (100 microM) caused a significant shift in the EC50 values for phenylephrine and methoxamine. Maximum responses in the DMV were significantly attenuated while those in the DSV were unaffected when total tension was considered. 4. Studies of the functional interactions of the DSV and the DMV with WB 4101 or 5-methylurapidil (5-MU) suggested the presence of alpha 1D-adrenoceptors in the DSV and alpha 1A-adrenoceptors in the DMV. The receptors inactivated by CEC in the DMV and DSV may represent some or all of the receptors with properties of alpha 1D and alpha 1A-receptors present in the two veins. Studies of radioligand binding interactions of these two antagonists with [3H] prazosin, were consistent with the presence of some alpha 1D-receptors in DSV and alpha 1A-receptors in DMV. These findings raise questions about the selectivity of CEC in differentiating alpha 1-adrenoceptor subtypes. 5. B-HT 920 caused contractions in the DSV smaller than those to the alpha 1-agonists but the maximum was not affected by CEC pretreatment. The EC50 values were shifted to the left after CEC. In radioligand binding studies, B-HT 920 competition for [3H] rauwolscine binding was not significantly affected by CEC pretreatment. 6. These results suggest the presence of unusual alpha-adrenoceptors in the DSV. In addition to alpha 2-adrenoceptors, receptors recognizing rauwolscine as well as prazosin, WB 4101, phenylephrine and methoxamine and susceptible to inactivation by CEC are present. They appear to be, in part, unusual alpha 1D-adrenoceptors. PMID- 8730752 TI - Inhibition by SK&F96365 of NO-mediated relaxation induced by Ca2(+) -ATPase inhibitors in rat thoracic aorta. AB - 1. We investigated the effect of SK&F96365, a putative inhibitor of receptor operated Ca2+ entry, on the endothelium-dependent, NO-mediated relaxation and cyclic GMP formation induced by Ca2(+)-ATPase inhibitors in rat thoracic aorta. 2. SK&F96365 inhibited cyclopiazonic acid or thapsigargin-induced relaxation and cyclic GMP formation mediated by a constitutive NO synthase, which is known to be activated by the Ca2+ that enters into the endothelial cells via plasma membrane Ca2+ channels subsequent to depletion of stored Ca2+ by Ca2(+)-ATPase inhibitors. 3. SK&F96365 also inhibited relaxation and cyclic GMP formation induced by acetylcholine, without affecting those induced by nitroprusside and A23187. 4. Ni2+ attenuated relaxation and cyclic GMP formation induced by cyclopiazonic acid and acetylcholine. 5. In contrast, the voltage-dependent Ca2+ channel blocker, nifedipine, did not affect the relaxation caused by Ca2(+)-ATPase inhibitors. 6. These results suggest that endothelium-dependent, NO-mediated relaxation of the arteries induced by Ca2(+)-ATPase inhibitors is triggered by the Ca2+ that enters into endothelial cells via receptor-operated channels (SK&F96365-sensitive channels) subsequent to depletion of stored Ca2+ as a result of inhibition of the Ca2(+)-ATPase (Ca2+ pump) of the stores. PMID- 8730753 TI - Vasodilator responses of rat isolated tail artery enhanced by oxygen- dependent, photochemical release of nitric oxide from iron-sulphur-nitrosyls. AB - 1. The vasodilator properties and photochemical decomposition of two synthetic iron-sulphur-nitrosyl clusters (cluster A: [Fe4S4(NO)4], tetranitrosyl-tetra-mu 3 sulphido-tetrahedro-tetrairon; and B:[Fe4S3 (NO)7]-1, heptanitrosyl-tri-mu 3 thioxotetraferrate(-1)) have been investigated. Experiments were carried out on isolated, internally-perfused segments of rat tail artery. 2. Bolus injections (10 microliters) of A or B ( > 0.25 mM) delivered into the internal perfusate generated sustained (or S-type) vasodilator responses, characterized by a persistent plateau of reduced tone due to NO released from clusters which enter and become trapped within endothelial cells. Clusters were therefore irradiated with visible laser light (lambda = 457.9 or 514.5 nm) either (a) in solution, while passing through a glass tube en route to the artery; or (b) when retained within the endothelium, by illuminating the artery directly during the plateau of an S-type response. Irradiation produced an additional vasodilator response, the magnitude of which depended upon wavelength and laser beam energy. 3. The nitric oxide synthase inhibitor, NG-monomethyl-L-arginine (100 microM), had no effect on light-induced vasodilator responses. However, they were (a) blocked entirely by adding oxyhaemoglobin (5 microM) to the internal perfusate; and (b) greatly enhanced by the enzyme superoxide dismutase (150 u ml-1). 4. Photolysis of cluster B was measured by absorption spectroscopy and by detecting NO released with an electrochemical sensor. The photochemical reaction was found to be oxygen dependent. The half-time for inactivation of cluster-derived NO was measured by interposing different lengths of tubing (i.e. time delays) between the photolysis tube and NO sensor. The steady-state probe current decayed exponentially with increasing delay time, with a t 1/2 of 21 s. The amplitudes of vasodilator responses of the tail artery also decreased exponentially by increasing the time delay (t 1/2 = 58 s). Superoxide dismutase (150 u ml-1) prevented this from happening, showing that "inactivation' of cluster-derived NO was caused by reaction with superoxide anions formed during photolysis. 5. We conclude that potentiation of vasodilator responses to iron-sulphur-nitrosyl clusters by visible light is due to an oxygen-dependent photochemical reaction which accelerates the release of ligated nitrosyl groups as free NO. Based on our measurements, we estimate that ca 100 pM NO is sufficient to produce a just detectable additional vasodilatation and that the ED50 dose is ca 3.7 nM. PMID- 8730754 TI - Pharmacological profile of TP-680, a new cholecystokininA receptor antagonist. AB - 1. The pharmacological characteristics of a newly developed serine derivative (R) 1-[3-(3-carboxypyridine-2-yl) thio-2-(indol-2-yl)carbonylamino]propionyl-4 diphenylmethyl- piperazine (TP-680), a cholecystokinin type A (CCKA) receptor antagonist, were studied and compared with those of MK-329 and loxiglumide. 2. TP 680 showed approximately 2 and 22 times greater selectivity for peripheral CCKA receptors relative to brain CCK (CCKB) receptors than MK-329 and loxiglumide, respectively, when IC50 values for inhibition of [125I]-CCK-8 binding in isolated acini and cerebral cortex were compared. 3. TP-680 was approximately 17 times less potent than MK-329, but was 106 times more potent than loxiglumide in inhibiting 100 pM CCK-8-stimulated amylase release from rat pancreatic acini. The antagonism produced by TP-680 was specific for CCK in that the effects of other receptor secretagogues or agents bypassing receptors were not altered. 4. TP-680 caused a parallel rightward shift of the dose-response curve for CCK-8-stimulated amylase release as did MK-329 and loxiglumide. However, in contrast to MK-329 and loxiglumide, TP-680 suppressed the maximal responses of CCK-8-induced amylase release in a concentration-dependent fashion, indicating that TP-680 is an unsurmountable antagonist. 5. Repeated washing of acini after a 30 min treatment with TP-680 restored the responsiveness but not the sensitivity, causing a residual inhibition on the action of CCK-8. 6. The addition of loxiglumide prior to or together with application of TP-680 protected CCK receptors from unsurmountable and irreversible antagonism by TP-680. 7. Our results indicate that TP-680 is a potent and the most selective CCKA receptor antagonist for the pancreas reported to date. PMID- 8730755 TI - Haemodynamic and cardiac effects of kinin B1 and B2 receptor stimulation in conscious instrumented dogs. AB - 1. Mongrel dogs were chronically instrumented with an intra-aortic catheter, a Konigsberg intraventricular pressure transducer and a Doppler flow probe around the left coronary artery. After ganglionic blockade with hexamethonium, the cardiovascular effects of bradykinin B1 and B2 receptor agonists, des-Arg9 bradykinin and bradykinin (BK), were investigated in the presence and absence of specific antagonists. The contribution of nitric oxide (NO) and prostanoids to the cardiovascular effects of kinins was also examined. 2. BK (1 microgram kg-1 min-1) and des-Arg9-BK (1 microgram kg-1 min-1) both given as a 2 min i.v. infusion, produced a significant decrease in mean arterial pressure (MAP, -34 +/- 4% for BK and -45 +/- 2% for des-Arg9-BK) and coronary vascular resistance (CVR, 37 +/- 5% for BK and -50 +/- 2% for des-Arg9-BK), without affecting cardiac contractility, left ventricular end diastolic pressure, and coronary velocity. BK caused a significantly greater decrease in MAP and CVR than des-Arg9-BK (P < 0.05). 3. Pretreatment with the B1 receptor antagonist, des-Arg9-[Leu8]-BK (25 micrograms kg-1) significantly inhibited the decrease in MAP and CVR produced by des-Arg9-BK but not by BK. Infusion of des-Arg9-[Leu8]-BK alone also induced a significant decrease in MAP and CVR (P < 0.05). In the presence of the B2 receptor antagonist, Hoe 140 (25 micrograms kg-1), only the decreases in MAP and CVR caused by BK were significantly reduced (P < 0.05). 4. Inhibition of NO synthase with N omega-nitro-L-arginine (L-NOARG, 45 mg kg-1) significantly (P < 0.05) prevented the decrease in CVR but not MAP induced by des-Arg9-BK, whilst responses to BK were not affected by L-NOARG pretreatment. Inhibition of prostanoid synthesis with indomethacin (25 mg kg-1) did not affect the reductions in MAP and CVR induced by des-Arg9-BK or BK. 5. In conclusion, i.v. des-Arg9-BK and BK administration induced reductions in MAP and CVR suggesting that in conscious instrumented dogs both B1 and B2 receptors are present and can affect systemic blood pressure and coronary resistance regulation. Our results also suggest that prostanoids are not involved in the vascular response to kinins and that coronary vascular B1 receptors are at least in part coupled to the release of NO. PMID- 8730756 TI - Evidence for a discrete UTP receptor in cardiac endothelial cells. AB - 1. We have examined the effects of various purine and pyrimidine nucleotides upon cells cultured from guinea-pig cardiac endothelium (CEC), and find the P2Y agonist 2-methylthioadenosine triphosphate (2MeSATP) to be a potent (EC50 = 85 +/ 10.2 nM) stimulator of increase in intracellular calcium concentrations, while uridine 5'-triphosphate (UTP) and adenosine 5'-triphosphate (ATP) are less potent but equipotent with one another (EC50s = 2.1 +/- 0.3 and 1.8 +/- 0.2 microM, respectively). 2. While the P2Y receptor exhibited rapid homologous desensitization, this had no effect upon subsequent responsiveness of CEC to either ATP or UTP. Effects of maximal concentrations of ATP and UTP were not only additive, but did not cross-desensitize. Responses to UTP (but not to ATP or 2MeSATP) were blocked by treatment with pertussis toxin (PTX); all three nucleotides appeared to liberate calcium from an intracellular pool. 3. Suramin (30 microM) significantly (P < 0.05) increased the EC50 for ATP-dependent increases in intracellular calcium (5.3 +/- 2.2 microM vs. 2.0 +/- 0.9 microM in the absence of suramin), while it completely blocked the response to 2MeSATP. Suramin had no effect upon responses to UTP at concentrations of 100 microM. 4. We conclude that in addition to the P2Y and P2U subtypes of the ATP receptor, an additional receptor responsive to UTP but exhibiting no affinity for purine nucleotides is present in CEC; this "pyrimidine receptor' liberates intracellular calcium via a G-protein, and may partly mediate the contractile response to UTP in the coronary vasculature. PMID- 8730757 TI - Role of nitric oxide in the development of tolerance and sensitization to behavioural effects of phencyclidine in mice. AB - 1. To determine whether nitric oxide (NO) was involved in tolerance and sensitization to the effects of phencyclidine (PCP), we examined NO synthase activity and the number of NADPH-diaphorase (NADPH-d)-positive cells in discrete brain regions of saline-, acute PCP- and repeated PCP-treated mice. We also investigated the effects of a NO synthase inhibitor, NG-nitro-L- arginine methyl ester (L-NAME), on the behavioural changes induced by repeated PCP treatment in mice. 2. Acute PCP (1, 3, and 10 mg kg-1, s.c.) treatment induced dose-dependent hyperlocomotion, motor incoordination and stereotyped behaviours, consisting of sniffing, head movement and ataxia in mice. 3. In mice treated repeatedly with PCP (1, 3, and 10 mg kg-1 day-1), s.c., once a day for 14 days), the sniffing, head movement, ataxia and motor incoordination induced by PCP were attenuated (indicating the development of tolerance to these behaviours), whereas the hyperlocomotion induced by PCP was potentiated (indicating the development of sensitization to hyperlocomotion). The development of tolerance and sensitization to PCP-induced behaviours in the repeated PCP-treated mice was more marked at the dose of 10 mg kg-1 day-1) than at other doses. 4. NO synthase activity in the cerebral cortex and cerebellum, but not in the striatum and hippocampus, was significantly decreased by acute PCP (10 mg kg-1) treatment in comparison with saline treatment, and such changes in activity in the cerebral cortex and cerebellum were reversed by repeated PCP treatment (10 mg kg-1 day-1). 5. The number of neurones containing NADPH-d reactivity in the cerebral cortex, nucleus accumbens, and striatum of acute and repeated PCP-treated mice showed no change in comparison with saline-treated mice. 6. Tolerance to PCP (10 mg kg-1 day-1) induced ataxia and motor incoordination was significantly attenuated by combined treatment with L-NAME (50 mg kg-1 day-1 i.p.). 7. Sensitization to PCP-induced hyperlocomotion was further enhanced by combined treatment with L-NAME (50 mg kg 1 day-1). However, NG-nitro-D-arginine methyl ester (D-NAME, 50 mg kg-1 day-1, i.p.), a less active enantiomer of L-NAME, had no effect, suggesting a stereospecific mechanism. 8. The PCP-induced behaviours in animals that had exhibited tolerance and sensitization to PCP (10 mg kg-1 day-1) were not influenced by acute L-NAME (5 and 50 mg kg-1, i.p.) or D-NAME (50 mg kg-1, i.p.) treatment. 9. These results suggest that NO may play an important role in the development, but not in the maintenance, of tolerance and sensitization to the effect of PCP in mice. PMID- 8730758 TI - Inhibition of the contraction of the ductus arteriosus to oxygen by 1 aminobenzotriazole, a mechanism-based inactivator of cytochrome P450. AB - 1. We have proposed that contractile tension of the ductus arteriosus is sustained by a cytochrome P450-linked mechanism acting as a limiting step in the synthesis of endothelin-1 (ET-1). In the present study, we have used the isolated ductus from near-term foetal lambs and guinea-pigs to investigate the effect on both muscle tone and ET-1 formation of 1-aminobenzotriazole (ABT), a suicide substrate for mono-oxygenase reactions. 2. ABT relaxed the lamb ductus at rest (2.5% O2) and during the oxygen contraction (15 to 95% O2). The effect was seen at 40 microM, and at 0.8 mM active tone was almost completely abolished. ABT (1 mM) also reversed the oxygen contraction in the guinea-pig ductus. 3. In the lamb ductus, the ABT response was not affected by removal of the endothelium or by treatment with 2.8 microM indomethacin (at 2.5% O2) and the ensuing contraction. 4. At both low and high concentration, ABT relaxed marginally, or not at all, the potassium-contracted (55 mM) ductus from either species. 5. ET-1 release from either the intact or endothelium-denuded lamb ductus tended to decrease in the presence of ABT (1 mM), whilst during the same treatment cyclic GMP content of the tissue remained unchanged. 6. We conclude that ABT relaxation is due to suppression of a contractile mechanism and not to activation of prostaglandin- and NO-mediated relaxing mechanisms. This contractile mechanism has a cytochrome P450-based mono-oxygenase reaction as a key component. PMID- 8730760 TI - Role of potassium channels in endothelium-dependent relaxation resistant to nitroarginine in the rat hepatic artery. AB - 1. In the presence of indomethacin (IM, 10 microM) and N omega-nitro-L- arginine (L-NOARG, 0.3 mM), acetylcholine (ACh) induces an endothelium-dependent smooth muscle hyperpolarization and relaxation in the rat isolated hepatic artery. The potassium (K) channel inhibitors, tetrabutylammonium (TBA, 1 mM) and to a lesser extent 4-aminopyridine (4-AP, 1 mM) inhibited the L-NOARG/IM-resistant relaxation induced by ACh, whereas apamin (0.1-0.3 microM), charybdotoxin (0.1-0.3 microM), iberiotoxin (0.1 microM) and dendrotoxin (0.1 microM) each had no effect. TBA also inhibited the relaxation induced by the receptor-independent endothelial cell activator, A23187. 2. When combined, apamin (0.1 microM) + charybdotoxin (0.1 microM), but not apamin (0.1 microM) + iberiotoxin (0.1 microM) or a triple combination of 4-AP (1 mM) + apamin (0.1 microM) + iberiotoxin (0.1 microM), inhibited the L-NOARG/IM-resistant relaxation induced by ACh. At a concentration of 0.3 microM, apamin + charybdotoxin completely inhibited the relaxation. This toxin combination also abolished the L-NOARG/ IM-resistant relaxation induced by A23187. 3. In the absence of L-NOARG, TBA (1 mM) inhibited the ACh-induced relaxation, whereas charybdotoxin (0.3 microM) + apamin (0.3 microM) had no effect, indicating that the toxin combination did not interfere with the L arginine/NO pathway. 4. The gap junction inhibitors halothane (2 mM) and 1 heptanol (2 mM), or replacement of NaCl with sodium propionate did not affect the L-NOARG/IM-resistant relaxation induced by ACh. 5. Inhibition of Na+/K(+)-ATPase by ouabain (1 mM) had no effect on the L-NOARG/IM-resistant relaxation induced by ACh. Exposure to a K(+)-free Krebs solution, however, reduced the maximal relaxation by 13% without affecting the sensitivity to ACh. 6. The results suggest that the L-NOARG/IM-resistant relaxation induced by ACh in the rat hepatic artery is mediated by activation of K-channels sensitive to TBA and a combination of apamin + charybdotoxin. Chloride channels, Na+/K(+)-ATPase and gap junctions are probably not involved in the response. It is proposed that endothelial cell activation induces secretion of an endothelium-derived hyperpolarizing factor(s) (EDHF), distinct from NO and cyclo-oxygenase products, which activates more than one type of K-channel on the smooth muscle cells. Alternatively, a single type of K-channel, to which both apamin and charybdotoxin must bind for inhibition to occur, may be the target for EDHF. PMID- 8730759 TI - Development of the model of rat isolated perfused heart for the evaluation of anthracycline cardiotoxicity and its circumvention. AB - 1. In order to develop a predictive model for the preclinical evaluation of anthracycline cardiotoxicity and the means of preventing it, we have studied the functional parameters of perfused hearts isolated from rats receiving repeated doses of several anthracyclines. 2. The anthracyclines studied were doxorubicin, epirubicin, pirarubicin and daunorubicin, and we also studied a liposomal formulation of daunorubicin (DaunoXome) and the co-administration of dexrazoxane (ICRF-187) and doxorubicin. 3. Anthracyclines were administered i.p. at equimolar doses corresponding to 3 mg kg-1 per injection of doxorubicin, every other day for a total of six doses. Dexrazoxane was used at the dose of 30 mg kg-1 per injection and was administered either 30 min before or 30 min after doxorubicin. We evaluated any general toxicity towards the animals as well as alterations of left ventricular contractility and relaxation ex vivo. 4. Epirubicin and daunorubicin were significantly less cardiotoxic than doxorubicin, and neither pirarubicin nor DaunoXome caused significant alterations in cardiac function. There was a direct relationship between the decrease in cardiac contractility or relaxation and anthracycline accumulation in the heart, evaluated after the same treatment schedule. 5. Dexrazoxane induced a significant protection against doxorubicin-induced cardiac toxicity when administered 30 min before doxorubicin, whereas this protection was ineffective when administered 30 min after doxorubicin. Direct perfusion of DaunoXome in isolated hearts of untreated animals resulted in a 12-fold reduction of the accumulation of daunorubicin in heart tissue as compared to the perfusion of free daunorubicin, and did not cause alterations in cardiac function at a dosage for which free daunorubicin induced major alterations. 6. The isolated perfused rat heart appears to be a valuable model for screening of new anthracyclines and of strategies for circumventing anthracycline cardiotoxicity. PMID- 8730761 TI - Societal self-defense: new laws to protect children from sexual abuse. PMID- 8730762 TI - The media and public awareness of child abuse and neglect: it's time for a change. PMID- 8730763 TI - The impact of news media on child abuse reporting. AB - This study explores the relationship between news stories on child abuse and neglect and reports of child abuse and neglect made to a mandated agency. Academic literature on crime news is reviewed to provide a context for interpretation. News stories from metropolitan daily newspapers were compared with child maltreatment reports made to mandated agencies on a local and national basis were surveyed over 25 years. The results suggested that both child maltreatment reports and news coverage increased over the period surveyed. However, rather than media stories increasing prior to increases in mandated reports and therefore contributing to the rise in reported cases, they appeared to increase at the same time. It is suggested that the initial cause of both increases may be national policy changes. More recent increases in child abuse reports may be due to economic downturns and other widespread societal changes rather than media attention. PMID- 8730764 TI - Reasonable cause: a qualitative study of mandated reporting. AB - Forty-nine volunteers (29 therapists and 20 child protection workers) were interviewed regarding their experiences with mandated reporting. The results indicated widespread disagreement between the groups about the nature of appropriate reports, the requirements of the law, and the process designed to protect children and help families. PMID- 8730765 TI - Threats of corporal punishment as verbal aggression: a naturalistic study. AB - This study examined firsthand observations of adults (N = 70) making public verbal threats to physically punish or hurt children. While many adults threatened the child with a "spanking," most used a variety of other terms, some euphemistic and some menacing, to label the threatened event. Angry shouting seemed rare, and swearing was almost nonexistent. Many threatening adults, however, also hit the child. In the course of threatening their children, adults typically attributed unshared responsibility for group problems to the child. They also normalized their own aggression by acting as though nothing unusual had happened. Perhaps for tactical reasons, most children also reacted as though there were nothing unusual about the threat. No one inside or outside the group intervened. It is argued that a complete understanding of verbal aggression against children requires an appreciation of the ongoing interaction in which episodes are embedded, and of the immediate social context in which episodes develop. Public places may be an important facilitative context because of the expectations associated with a parent's public persona. PMID- 8730766 TI - Moderating effects of physical abuse and perceived social support on the potential to abuse. AB - This study investigated the moderating effects of perceived prepubertal social support on the relationship between prepubertal childhood physical abuse and child abuse potential as well as on the relationship between a childhood history of abuse and conflict and depth in adult relationships. Three hundred and sixty nine undergraduate men and women were classified as abused or nonabused based on their responses to the Childhood History Questionnaire. Multiple regression analyses revealed interactions of social support and a childhood history of physical abuse on both the potential to abuse and the quality of participants' relationships with their parents. Differences between abused and nonabused individuals were obtained only under conditions of low perceived social support. When social support was perceived to be high, abused and nonabused individuals did not differ in the potential to abuse or in the ratings of the depth of their relationships with parents. Implications of these findings for the intergenerational transmission of abuse are discussed. PMID- 8730767 TI - Sexual abuse evaluations in the emergency department: is the history reliable? AB - The authors assessed agreement on perpetrator identification obtained at two interviews of child victims of sexual abuse. We reviewed charts for 141 children who had undergone both a screening interview by an emergency department physician and an investigative interview by an interdisciplinary team specializing in child sexual abuse evaluation. For 107 (76%) cases, information was consistent; for instance, identification occurred at both interviews or at neither interview. Of the 38 children not identifying the perpetrator at the screening interview, 17 (45%) made disclosure at the investigative interview. Children who made disclosure only at the investigative interview were more likely to have refused to speak to the physician in the emergency department (p = .001). No significant differences were found when data for interview site and perpetrator identification were analyzed by gender of child, time interval between assault and screening interview, time interval between the two interviews, relationship of the alleged perpetrator to the child, performance of the investigative interview on an inpatient versus outpatient basis, and report of genital contact. In conclusion, perpetrator identification obtained during emergency department screening interviews usually agreed with information obtained at the subsequent investigative interview. The investigative interview was most helpful in identification of perpetrators when the child was not the source of the history obtained in the emergency department. PMID- 8730768 TI - Characteristics of childhood sexual abuse among female survivors in therapy. AB - Characteristics of the abuse experience by 135 women entering an outpatient treatment program for survivors of childhood sexual abuse were assessed utilizing a structured clinical interview. Areas assessed included: nature, frequency and duration of the abuse; participant's age at onset of abuse; perpetrator characteristics; childhood physical abuse experiences; and circumstances leading to abuse cessation. Participants who had been molested by more than one perpetrator were administered questions about the abuse committed by each perpetrator separately. Abuse by the first individual to molest a participant was found to be more likely to be incestuous, of higher frequency, and more extensive and invasive than that by later perpetrators. Survivors seeking therapy indicated experiencing considerably more severe abuse, at a younger age, for longer duration, and at the hands of more perpetrators than previously reported in the literature on nonclinical samples of survivors. PMID- 8730769 TI - Exploring new territory: police organizational responses to child sexual abuse. AB - Police response to the escalating numbers of child sexual abuse cases has led to the development of a wide range of organizational models within both police departments and the interagency context. This paper compares the results of different models of police intervention in New South Wales, Australia. The data suggest that specialist police units in which police manage the full investigation provide a more comprehensive service than those where the police response is fragmented between units within the police force. PMID- 8730770 TI - The helping alliance in work with families where children have been abused or neglected. PMID- 8730771 TI - Preserving families at risk of child abuse and neglect: the role of the helping alliance. AB - Family preservation services are increasingly employed to decrease the use of foster care and preserve maltreating families at high risk of child placement. First studies of family preservation services appeared to support their effectiveness in achieving these goals. However, questions are raised increasingly about their effectiveness, particularly with those families whose functioning is impaired by psychopathology and substance abuse. The time has come to seek fuller understanding of family preservation services and their differential successes. In this paper, we draw on current research in the field of psychotherapy--specifically on studies of the helping alliance and its relationship to treatment outcome--to purpose a new look at family preservation services and to inform of treatment of high-risk families. PMID- 8730772 TI - Family functioning in neglectful families. AB - Family functioning in 103 neglectful and 102 non-neglectful low-income families is examined using self-report and observational measures. Neglectful mothers reported their families as having more family conflict and less expression of feelings, but not less cohesive. Ratings of observed and videotaped family interactions indicated neglect families were less organized, more chaotic, less verbally expressive, showed less positive and more negative affect than comparison families. However, there were wide differences on measures of functioning among neglect families. Three distinct types of neglectful family functioning are identified and interventions for each type are suggested to improve parental-family functioning. PMID- 8730773 TI - Role of neurotrophins and trk receptors in the development and maintenance of sensory neurons: an overview. AB - The neurotrophins are a family of polypeptide neuronal growth factors related to the prototypical neurotrophic factor, nerve growth factor (NGF). In mammals this gene family encompasses NGF, brain-derived neurotrophic factor (BDNF) and neurotrophins-3 and -4/5, (NT-3, NT-4/5). The neurotrophins initiate signal transduction in responsive cells by ligand induced dimerization and activation of one of the Trk family of receptor tyrosine kinases; NGF being specific for TrkA, BDNF and NT-4/5 for TrkB, and TrkC the preferred receptor for NT-3. In accord with differential patterns of distribution of Trk receptors in peripheral ganglia, the neurotrophins show both distinct and overlapping specificity towards subpopulations of sensory neurons of both neural crest and neural placode origin. In vitro and in vivo studies, and transgenic mice baring targeted null mutations of the neutrophin genes have established that BDNF, NT-3 and NT-4/5, like NGF, play critical roles as classical target-derived survival factors for subclasses of developing sensory neurons. However, much broader effects of neurotrophins on sensory neurons are now evident, including paracrine and autocrine actions on neuroblast proliferation, phenotypic differentiation, and survival and regeneration in the adult. This article provides an overview of the discovery and properties of the neurotrophin family, their receptors and their actions and specificity for both distinct and overlapping subpopulations of spinal and cranial sensory neurons. PMID- 8730774 TI - The role of neurotrophins in development of neural-crest cells that become sensory ganglia. AB - A fundamental issue of neural-crest ontogeny is understanding how different types of cells are created at the right time and in the correct numbers. Sensory ganglia are among the many derivatives of the vertebrate neural crest. Their proper formation requires the regulation of several processes such as cell fate specification, proliferation, survival, and terminal differentiation. The timescale of the occurrence of processes involved in the regulation of cell number and identity, coincides with key morphogenetic events such as cell migration, homing and gangliogenesis. To gain insight into these processes, we characterized the cellular basis of metameric migration of neural-crest cells and of consequent ganglion organization, which are imposed by intrinsic differences within rostral and caudal sclerotomal compartments. We also established a transient requirement for neural tube-derived factors in regulating the proliferation, survival and differentiation of prospective DRG cells. Additionally, we showed that cooperation between the mesodermal cells and the neural tube is necessary for modulating cell number in the nascent ganglia. BDNF, NT-3 and basic FGF were found to mediate this environmental signalling. All the above factors display neurogenic activity for a subset of early-committed sensory neuron progenitors. This observation raises the possibility of an early redundancy in the response of individual neural-crest progenitors to distinct factors. This overlap in responsiveness progressively disappears upon the colonization of specific ganglionic sites and the subsequent establishment of selective innervation patterns by post-mitotic sensory neurons. PMID- 8730775 TI - Roles of neurotrophin-3 during early development of the peripheral nervous system. AB - The neurotrophins are structurally related proteins regulating cell numbers in the developing vertebrate nervous system. They are necessary survival factors preventing the death of specific neuronal populations. Previous experiments have indicated that the administration of nerve growth factor or of brain-derived neurotrophic factor during the formation of sensory ganglia and of target innervation increases the number of neurons by preventing normally occurring neuronal death. These results support the view that during development, neuronal numbers are adjusted to the size of the target tissue by the release of limiting amounts of neurotrophins. However, increasing the levels of neurotrophin-3 during the formation of sensory ganglia results in a marked decrease in neuronal numbers, possibly as a consequence of premature cessation of sensory neuroblast proliferation. In sympathetic ganglia, the application of neurotrophin-3 during the formation of the sympathetic chain causes cell numbers to increase, a result also observed following the application of nerve growth factor. It thus appears that neurotrophin-3 and nerve growth factor can regulate cell numbers well before the period of target-derived control, and that neurotrophin-3 affects neuronal numbers in sensory and sympathetic ganglia in opposite ways. PMID- 8730776 TI - The neurotrophic hypothesis: where does it stand? AB - In the developing peripheral nervous system many neurons die shortly after their axons reach their target fields. This loss is thought to match the number of neurons to the size and requirements of their target fields because altering target field size before innervation affects the number of neurons that survive. The neurotrophic hypothesis provides an explanation for how target fields influence the size of the neuronal populations that innervate them. This hypothesis arose from work on nerve growth factor (NGF), the founder member of the neurotrophin family of secreted proteins. Its principal tenet is that the survival of developing neurons depends on the supply of a neurotrophic factor that is synthesized in limiting amounts in their target fields. The neurotrophic hypothesis has, however, been broadened by the demonstration that multiple neurotrophic factors regulate the survival of certain populations of neurons. For example, some neurons depend on several different neurotrophic factors which may act concurrently or sequentially during target field innervation. In addition, there are aspects of neurotrophin action that do not conform with the classic neurotrophic hypothesis. For example, the dependence of some populations of sensory neurons on particular neurotrophins before significant neuronal death takes place raises the possibility that the supply of these neurotrophins is not limiting for survival at this stage of development. There is also evidence that at stages before and after sensory neurons depend on target-derived neurotrophins for survival, neurotrophins act on at least some sensory neurons by an autocrine route. Yet despite the growing wealth of information on the multiple roles and modes of action of neurotrophic factors, the neurotrophic hypothesis has remained the best explanation for how neuronal target fields in the developing peripheral nervous system regulate their innervation density. PMID- 8730777 TI - Dorsal root ganglion neurons require functional neurotrophin receptors for survival during development. AB - Neurotrophins are the most profound known regulators of survival in the developing peripheral nervous system. Within dorsal root ganglia, the signalling receptors for the different members of the neurotrophin family are distributed in distinct patterns suggesting regulation of different functional classes of sensory neurons. Abnormalities observed in neurotrophin receptor mutant mice have confirmed this idea. Both trkA (-/-) and trkC (-/-) mice have striking neurological defecits referrable to subpopulations of DRG neurons which have distinct axon projections in the periphery. These results thus generalize concepts of dependence on target-derived factors based on extensive work with the prototypical neurotrophin, nerve growth factor. Further analysis of these animals also provides evidence for more complex developmental mechanisms including dependence on locally synthesized neurotrophins at early developmental stages and plasticity of neurotrophin receptor expression. PMID- 8730778 TI - Neurotrophins and the specification of neuronal phenotype. AB - Nerve growth factor, brain derived neurotrophic factor and neurotrophin-3 all influence sensory neurons derived from the dorsal root ganglia. Traditionally these neurotrophins have been thought of as survival factors for sensory neurons during their development. Recent evidence from experiments where the in vivo levels of these proteins has been manipulated indicates that they may influence the development of specific sensory neuron phenotypes. In this review these experiments are discussed in relation to the mechanisms by which neurotrophins could influence the phenotypic fate of sensory neurons. The first mechanism requires that when a neuron becomes dependent for survival on a neurotrophin the availability of the factor simply influences the number of neurons surviving with a certain modality. This model requires that neurotrophin responsiveness is a determinant of the possible modalities that the neuron may acquire. The second mechanism requires that the availability of a given neurotrophin influences how many neurons can differentiate into different sensory neuron phenotype independent of survival. The available experimental data is discussed in relation to these two models. PMID- 8730779 TI - Expression of the trk family of neurotrophin receptors in developing and adult dorsal root ganglion neurons. AB - Expression of trk receptors is a major determinant of neurotrophin responsiveness of sensory neurons. Although it has been apparent for some time that subpopulations of dorsal root and trigeminal ganglion neurons respond in vitro to each of the members of the neurotrophin family, the extent to which functionally distinct subclasses of sensory neurons are dependent on the actions of different neurotrophins for their development and function remains an active area of investigation. One step towards elucidating the role of various neurotrophins in development and function of sensory neurons has been to examine the distribution of trk receptors on sensory neurons. These studies have clearly revealed that members of the trk family are differentially expressed in functionally distinct populations of both developing and mature sensory neurons and, further, have provided evidence consistent with a shift in neurotrophin responsiveness during the development of sensory neurons. PMID- 8730780 TI - Non target-derived roles of the neurotrophins. AB - The hypothesis that target-derived neurotrophic factors are essential for the survival, differentiation and maintenance of sensory, sympathetic and motor neurons has been well supported by analysis of mice bearing null mutations in the neurotrophins and their receptors. However, the localization of brain-derived neurotrophic factor (BDNF) in a population of dorsal root ganglia (DRG) sensory neurons (Ernfors et al. 1990b; Ernfors & Persson 1991; Schecterson & Bothwell 1992) suggested the additional possibility that BDNF could act in a paracrine or autocrine manner to mediate neuronal survival. We tested this hypothesis in cultured adult DRG neurons, which survive as single cells in microwells in the absence of added trophic factors (Lindsay 1988). About 35% of these neurons were specifically killed by BDNF antisense oligonucleotide administration in a dose dependent manner, with no effect of sense oligonucleotides. Antisense administration was accompanied by an 80% decrease in BDNF protein levels over the first 24 h of treatment (Acheson et al. 1995). The BDNF autocrine loop that we propose to be present in sensory neurons may be representative of a broader phenomenon in the nervous system as a whole, where the balance of neurotrophic support may shift during development from target-derived to paracrine or autocrine modes. Perhaps as a consequence of this developmental shift, the survival of both peripheral nervous system (PNS) and central nervous system (CNS) neurons in the adult is less affected by axotomy or target removal when compared to their response during development. PMID- 8730781 TI - Neurotrophins and nerve injury in the adult. AB - A role for neurotrophins in mature primary sensory neurons persists, extending beyond that of promoting survival during development, to one of maintaining phenotypic and functional properties. Many adaptive changes that occur after peripheral axotomy and in axonal repair are believed to be influenced by altered availability of neurotrophic molecules to the neuron in this state. Indeed, administration of exogenous nerve growth factor counteracts many degenerative changes observed in the subpopulation of axotomized neurons which are nerve growth factor-responsive. Current efforts focus on defining actions of other neurotrophins (brain-derived neurotrophic factor, neurotrophin-3 and neurotrophin 4/5) in nerve injury and repair, and the intracellular pathways involved. Knowledge gained from work focusing on nerve growth factor and neurotrophin-3 in supporting maintenance or modulation of aspects of the differentiated state of adult primary sensory neurons is discussed. PMID- 8730782 TI - NGF as a mediator of inflammatory pain. AB - The chapter reviews some of recent evidence which suggests that one neurotrophin, nerve growth factor (NGF), is a peripherally produced mediator of some persistent pain states, notably those associated with inflammation. The evidence for this proposal is as follows. 1. The endogenous production of NGF regulates the sensitivity of nociceptive systems. Behavioural and electrophysiological studies have shown that sequestration of constitutively produced NGF leads to decrease nociceptor sensitivity. 2. In a wide variety of experimental inflammatory conditions NGF levels are rapidly increased in the inflamed tissue. 3. The high affinity NGF receptor, trkA, is selectively expressed by nociceptive sensory neurons particularly those containing sensory neuropeptides such as substance P and CGRP. 4. The systematic or local application of exogenous NGF produces a rapid and prolonged behavioural hyperalgesia in both animals and humans. Exogenous NGF has also been found to activate and sensitize fine calibre sensory neurons. 5. In a number of animal models, much of the hyperalgesia associated with experimental inflammation is blocked by pharmacological "antagonism' of NGF. The mechanisms by which NGF up-regulation in inflamed tissues might lead to sensory abnormalities is also discussed. In particular, evidence is reviewed which suggests that increased NGF levels leads to both peripheral sensitization of nociceptors and central sensitization of dorsal horn neurons responding to noxious stimuli. PMID- 8730783 TI - Phenotypic modification of primary sensory neurons: the role of nerve growth factor in the production of persistent pain. AB - Inflammation results in an early and maintained elevation in nerve growth factor (NGF) levels in inflamed tissues. Neutralizing the action of the increased NGF with specific anti-NGF antibodies substantially diminishes inflammatory hypersensitivity, indicating that this neurotrophin is a key mediator in the production of inflammatory pain. The hyperalgesic actions of NGF may in part be the consequence of an increase in sensitivity of the peripheral terminals of high threshold nociceptors either as a result of a direct action of NGF on trkA expressing sensory fibres or indirectly via the release of sensitizing mediators from trkA expressing inflammatory cells and postganglionic sympathetic neurons. NGF is also, however, retrogradely transported in sensory neurons to the dorsal root ganglion where it alters transcription of a number of proteins and peptides. This chapter reviews evidence suggesting that an NGF-mediated modification of gene expression in the dorsal root ganglion during inflammation is central to the pathophysiology of persistent pain. The phenotype changes produced by NGF during inflammation include elevation of neuropeptides which may amplify sensory input signals in the spinal cord and augment neurogenic inflammation in the periphery and the upregulation of growth related molecules which may lead to a hyperinnervation of injured tissue by promoting terminal sprouting. PMID- 8730784 TI - Neurotrophins and peripheral neuropathy. AB - Endogenous nerve growth factor (NGF) levels were studied in patients with nerve trauma, diabetes mellitus and leprosy, the most common causes of human peripheral neuropathy. In diabetics, there was an early length-dependent dysfunction of small-diameter sensory fibres, with depletion of skin NGF and the sensory neuropeptide substance P. The NGF depletion correlated significantly with decreased skin axon-reflex vasodilatation, which is mediated by small sensory fibres at least partly via substance P release. Immunostaining showed depletion of NGF in keratinocytes in diabetic skin. In injured nerves, NGF levels were reduced when compared to intact nerve, except acutely distal to injury; NGF immunostaining was seen in Schwann cells in distal segments, including neuromas. NGF levels were decreased in leprosy-affected skin and nerve. The role of neurotrophins in the rational treatment of human neuropathies is discussed e.g. loss of nociception and axon-reflex vasodilatation contribute to skin ulceration, a major and serious complication, for which NGF may provide prophylaxis. PMID- 8730785 TI - Neurotrophins and peripheral neuropathy. AB - The most common form of peripheral neuropathy is that associated with diabetes mellitus. In rodent models of diabetes there are expression deficits in nerve growth factor (NGF) and in its high-affinity receptor, trkA, leading to decreased retrograde axonal transport of NGF and decreased support of NGF-dependent sensory neurons, with reduced expression of their neuropeptides, substance P and calcitonin gene-related peptide (CGRP). Treatment of diabetic rats with intensive insulin normalized these deficits and treatment with exogenous NGF caused dose related increases, giving levels of NGF and neuropeptides which were greater than those of controls. Neurotrophin-3 (NT-3) mRNA was also deficient in leg muscle from diabetic rats and administration of recombinant NT-3 to diabetic rats increased the conduction velocity of sensory nerves without affecting motor conduction velocity. These findings implicate deficient neurotrophic support in diabetic neuropathy and suggest that its correction should be a paramount therapeutic target. PMID- 8730786 TI - Neurotrophins and sensory neurons: role in development, maintenance and injury. A thematic summary. PMID- 8730787 TI - The systematics of North American Daphnia (Crustacea: Anomopoda): a molecular phylogenetic approach. AB - Despite extensive studies on the ecology and evolution of the freshwater microcrustacean Daphnia, there is little understanding of the evolutionary history of the genus. Past attempts at reconstructing phylogenetic relationships among Daphnia species have been highly controversial, mainly because of the poor taxonomy of the genus. However, following a revised taxonomy of the daphniid fauna of North America, we conducted a comprehensive appraisal of systematic relationships within the genus through the analysis of sequence diversity in 503 b.p. of the 12S rRNA gene of the mtDNA. The large sequence divergence among its 34 North American members indicates that the genus Daphnia originated during the Mesozoic, even though many lineages exhibit extreme morphological stasis. Results from both cladistic and phenetic analyses indicate the presence of three subgenera comprised of 15 species complexes. Only four of these lineages have shown active speciation over the past 3 Ma, suggesting that cladogenesis in the genus has been constrained. Our study also reveals that interspecific hybridization occurs between taxa which show very large sequence divergence (up to 14%), suggesting that reproductive isolation within the genus evolves slowly. PMID- 8730788 TI - The effect of riluzole on post-traumatic spinal cord injury in the rat. AB - This study evaluated treatment with riluzole, a neuroprotective agent, following thoracic spinal cord compression in the rat. Animals received riluzole (2 mg kg 1) or vehicle twice daily for 10 days following the trauma. Motor deficits, somatosensory evoked potentials (SEP) and lesion histology were evaluated. Although paralysis was seen following trauma, seven of 10 animals receiving riluzole recovered motor function and nearly normal behaviour, unlike animals receiving vehicle. Trauma dramatically disturbed SEPs with falls in amplitude and increases in latency. After riluzole SEP returned towards pre-injury levels, while untreated animals showed no recovery. Morphological studies revealed significant (53%) reduction in the degree of spinal cord infarcted after riluzole treatment. PMID- 8730789 TI - Acute or chronic lithium does not affect agonist-stimulated inositol trisphosphate formation in rat brain in vivo. AB - Rats were given lithium either acutely by s.c. injection (4 m eq kg-1) or chronically by including 0.2% LiCl in their diet for 3 weeks. Microdialysis probes were inserted into the cortex or hippocampus, using re-usable guides, and perfused with artificial CSF. Fractions were collected beginning 18 h after the end of treatment and were analysed for inositol trisphosphate (IP3). Neither acute nor chronic treatment affected basal levels of IP3 or stimulation of IP3 formation by either carbachol or noradrenaline in the hippocampus. Similarly, neither basal nor carbachol-stimulated IP3 levels in rat cortex were affected by acute Li administration. It would appear that the reductions in these parameters previously reported by other workers using brain slices were due to inositol depletion occurring at the stage of brain slice preparation. The inositol depletion hypothesis for the mechanism of action of lithium does not therefore appear to be supported by in vivo evidence. PMID- 8730790 TI - 1-Aminocyclopropane-carboxylic acid reduces NMDA-induced hippocampal neurodegeneration in vivo. AB - The effect of systemic treatment with 1-aminocyclopro-panecarboxylic acid (ACPC), a partial agonist at the glycine site of the NMDA receptor, on convulsions and neurodegeneration induced by intrahippocampal injection of NMDA was investigated in mice. Five days after intrahippocampal NMDA infusion, 80-100% pyramidal cell death was observed in the CA1 region of the hippocampus. Pretreatment with ACPC prevented the lethal effects of NMDA and significantly reduced seizure induction. ACPC reduced cell death to 40% of that induced by a dose of NMDA (6 nmol) that damaged 80% of hippocampal CA1 neurones in untreated animals. These findings provide further evidence that ACPC can reduce NMDA receptor function in vivo and suggest that partial agonists at the glycine site of the NMDA receptor complex may be useful anticonvulsant and neuroprotective agents. PMID- 8730791 TI - Time course and role of extracellular Ca2+ in veratridine-induced glutamate release. AB - The veratridine-induced release of glutamate in the striatum of anaesthetized rats was studied using regional brain microdialysis to elucidate the role of extracellular Ca2+ and to investigate the exact time course of the glutamate response. In a first series of experiments with Ca(2+)-depletion starting simultaneously with the veratridine pulse no significant differences were found under Ca(2+)-replete and Ca(2+)-free conditions. Strikingly, under both conditions glutamate efflux followed a biphasic time course. When Ca2+ depletion preceded veratridine stimulation a significant inhibition of the initial glutamate release was found, indicating that the first glutamate portion originates from Ca(2+)-dependent vesicular exocytosis. The nature of the second and longer lasting phase of glutamate efflux is not clear, but it may be mediated by reversal of the glutamate uptake system of the plasma membrane located both on neurones and on astrocytes. This points to a possibly important contribution of glial cells to glutamate overflow under pathological conditions and should be subject to further investigations. PMID- 8730793 TI - Monkey saccadic latency and pursuit velocity show a preference for upward directions of target motion. AB - Saccadic latency was studied as a function of the direction of sudden target displacements (steps) and of subsequent smooth target motion (ramps) in Macaca fascicularis. The monkey fixated a central spot that suddenly changed its position and then moved constantly at 10 deg s-1, thus eliciting initial saccades and subsequent pursuit eye movements (recorded by a magnetic search-coil technique). Latencies for initial saccades differed markedly in the vertical axis, being shorter in upward than downward directions for both step and ramp components of target motion. Saccadic latency was also related to the mean pursuit velocity, indicating that the oculomotor system accounts for the direction of step and ramp components of target motion in an integrative way. PMID- 8730792 TI - Effects of tactile interference stimulation on somatosensory evoked magnetic fields. AB - Tactile stimulation of the hand interferes with somatosensory evoked brain responses following electrical median nerve stimulation. This effect was studied in eight normal subjects by magnetonecephalography (MEG). When stimulation was applied to the hand ipsilateral to the stimulated nerve, only the third response (3M) was enhanced in five subjects, but other responses were attenuated in all subjects. These interference effects were probably due to interactions in areas 3b and 1. After stimulation of the contralateral hand, only the second response (2M) was enhanced in six subjects. This effect was probably due to the intracerebral interactions mediated through the corpus callosum. PMID- 8730794 TI - ERPs to pitch changes: a result of reduced responses to standard tones in rabbits. AB - Event-related potentials (ERPs) were recorded in rabbits when pitch deviant tones occurred in a series of standard tones (oddball situation). In control recordings, the deviant tones were presented without the standard tones (deviant alone situation). In the oddball situation, significant difference ERPs (deviant ERPs-standard ERPs) could be found in the hippocampal and cerebellar recordings but not in the visual cortex. All the ERPs to the deviant stimuli observed in the oddball situation were also present in the deviant-alone situation. The difference ERPs were therefore based on reduced responses to the standards. The results are discussed in the context of a mismatch negativity (MMN) in humans. PMID- 8730795 TI - Muscarinic receptor subtypes in the medial preoptic area and sleep-wake cycles. AB - To clarify which muscarinic receptor subtype(s) mediate changes in sleep and cortical temperature (Tcort) induced by carbachol microinjections into the medial preoptic area (MPA), pirenzepine, tripitramine and +/- p < > -fluorohexahydro sila-difenidol (p-F-HHSiD), which are highly selective muscarinic M1, M2 and M3 antagonists, respectively, were microinjected into the MPA of rats. Whereas pirenzepine (3.45 and 7.08 nmol) and p-F-HHSiD (3.90 and 7.80 nmol) were without effect, tripitramine (0.67 and 3.37 nmol) enhanced wakefulness, decreased slow wave and desynchronized sleep, and raised Tcort with the higher dose. The data suggest that in the MPA only M2 muscarinic subtypes may be functionally important in mediating the cholinergic effects on sleep and thermoregulation. PMID- 8730796 TI - L-AP4 inhibits high voltage-activated Ca2+ currents in pyramidal cortical neurones. AB - We tested the ability of L-AP4 to modulate high voltage-activated (HVA) calcium (Ca2+) currents in pyramidal neurones acutely isolated from the adult rat (4-8 weeks). Whole cell recordings, with barium (Ba2+) ions as the charge carrier, were performed. L-AP4 reduced HVA Ca2+ conductances in 86% of the recorded cells. Saturating concentrations of L-AP4 inhibited about 21% of the current (+/- 8.3%, n = 8), although great variability was observed. Interestingly, low micromolar concentrations of (1S,3R)-1-aminocyclo-pentane-1,3-dicarboxylic acid (1S,3R-ACPD) and (2S,3S,4R)-alpha-(carboxy-cyclopropyl)-glycine (1-CCGI) had weaker effects than L-AP4. MAP4 fully antagonized the L-AP4-mediated reduction of HVA Ca2+ currents. These findings suggest the involvement of the AP4-sensitive receptor in the control of both cellular excitability and transmitter release in rat neocortical neurones. PMID- 8730797 TI - Apoptosis and c-Jun in the thalamus of the rat following cortical infarction. AB - Cortical infarction produces secondary neuronal damage in the thalamus. In this study we examined the thalamus of the rat following 2 h occlusion of the middle cerebral artery (MCA), and found degeneration and gliosis in the ipsilateral ventropostero-medial thalamic nucleus in those rats that showed cortical infarction 7 and 14 days after occlusion. This was accompanied by isolated cells with fragmented DNA, as revealed by in situ labelling of nuclear DNA fragmentation, and showing morphological features of apoptosis, i.e. chromatin condensation, extreme nuclear shrinkage and apoptotic bodies. In addition, cells immunoreactive for c-Jun showing morphological signs of apoptosis were observed. These results provide evidence of apoptosis in the ipsilateral thalamus following cortical infarction, and suggest that c-Jun is involved in this process. PMID- 8730798 TI - Over-expression of HSP-70 protects astrocytes from combined oxygen-glucose deprivation. AB - Pretreatment by a sublethal insult is associated with induction of stress proteins and with protection from subsequent injury. Heat pretreatment protects the brain from subsequent ischemia, and is shown here to protect primary astrocyte cultures from subsequent oxygen-glucose deprivation. To determine whether the expression of a single stress protein, HSP-70, could account for much of this protection, we expressed HSP-70 or beta-galactosidase in astrocytes using retroviral vectors. Only 12% of astrocytes expressing HSP-70 died after 7 hours of oxygen-glucose deprivation compared to 65% of astrocytes expressing beta galactosidase and 82% of normal astrocytes. Our data provide direct evidence that selective expression of HSP-70 enhances the survival of astrocytes challenged with heat or oxygen-glucose deprivation. PMID- 8730799 TI - The HIV-1 envelope protein gp120 induces neuronal apoptosis in hippocampal slices. AB - The HIV-1 envelope protein gp120 produces neuronal cell damage in primary cultures of a variety of cell types including hippocampal and retinal ganglion cell neurons. The properties of primary cell cultures are, however, often markedly different from those of cells living in their normal environment. We now report that gp120 induces widespread chromatin condensation and lesions in pyramidal granular neurones and in interneurones of rat hippocampal organotypic slice cultures. This damage is clearly of an apoptotic (programmed cell death) type. The use of an in vitro organized structure will enable the molecular and cellular mechanism of action of gp120 to be examined in conditions which are particularly suitable and relevant to the in vivo situation. PMID- 8730800 TI - Up-regulation of Bcl-2 is associated with neuronal DNA damage in Alzheimer's disease. AB - Cell death and neurofibrillary tangle formation are prominent features of Alzheimer's disease (AD). It has been suggested that DNA damage may reflect neuronal vulnerability. In this context, the Ced homologue Bcl-2 is able to repress a number of cell death programs. Recently we found both numerous nuclei exhibiting DNA damage within neurons in the AD brain and increases in Bcl-2 immunoreactivity. In this study, we examined the relationship between Bcl-2 expression and nuclear DNA damage or tangle formation. Nuclei exhibiting DNA damage were associated with an up-regulation of Bcl-2 expression, whereas tangle bearing neurons were associated with a down-regulation of Bcl-2 expression. PMID- 8730801 TI - Glia conditioned medium protects fetal rat midbrain neurones in culture from L DOPA toxicity. AB - L-DOPA kills dopamine neurones in culture but is the most effective drug for the treatment of Parkinson's disease, where it exhibits no clear toxicity. While glial cells surround and protect neurones in vivo, neurones are usually cultured in vitro in the absence of glia. We treated fetal midbrain rat neurones with L DOPA, mesencephalic glia conditioned medium (CM) and L-DOPA + CM. L-DOPA reduced the number of tyrosine hydroxylase-positive (TH+) cells and [3H]DA uptake, and increased quinone levels. L-DOPA + CM restored [3H]DA uptake and quinone levels to normal, and increased the number of TH+ cells and terminals to 170% of control. CM greatly increased the number of TH+ cells and [3H]DA uptake. Mesencephalic glia therefore produced soluble factors which are neurotrophic for dopamine neurones, and which protect these neurones from the toxic effects of L DOPA. PMID- 8730802 TI - Protective effect of vagus nerve stimulation on forebrain ischaemia in gerbil hippocampus. AB - The left vagus nerve was stimulated during transient forebrain ischaemia in gerbils. The animals were exposed to 3 min of forebrain ischaemic insult at 37.5 degrees C. On day 5 post-ischaemia, the animals were perfusion-fixed for qualitative and quantitative histopathological analyses. High current stimulation of the vagus nerve inhibited ischaemic neuronal damage in the hippocampal CA1 sector (p < 0.01), but low current stimulation did not (p < 0.01). These effects might have been due to inhibition of the effects of excitatory amino acids during ischaemia. These results indicate that vagus nerve stimulation might be protective to neurones subjected to ischaemic insult. PMID- 8730803 TI - Increased food intake and body weight gain after lateral hypothalamic dopaminergic cell implantation. AB - To examine our hypothesis that dopamine activity in the lateral hypothalamic area (LHA) may play a role in enhancing the process of eating, a fetal cell suspension of predominantly dopaminergic cells was bilaterally transplanted into the LHA of study rats via direct injection; controls had carrier medium injection. Thereafter, mean daily food intake was 1 g per day greater in dopaminergic cell transplanted rats vs. controls for each day of the 10-week observation period. This resulted in a significantly greater cumulative body weight gain in study rats vs. controls (386 +/- 5.1 g vs. 354 +/- 3.8 g, respectively). On sacrifice at the end of the study, transplanted cells in the LHA were viable. Our data suggest that bilateral LHA dopaminergic cell transplant which presumably resulted in chronically and persistently enhanced dopaminergic activity in the LHA is associated with overeating and consequently, an excess weight gain. PMID- 8730804 TI - Dopamine modulates the afterhyperpolarization in neostriatal neurones. AB - Intracellular techniques were used to study the actions of dopaminergic D1 agonists on the afterhyperpolarization (AHP) that follows action potentials in rat neostriatal neurones. Dopamine or Cl-APB (10 microM), or 1-10 microM 6-Cl-PB all increased AHP amplitude. This effect was blocked by 1 microM SCH-23390, a D1 antagonist, but not by 1 microM sulpiride, a D2 antagonist. Both 500 microM dibutyryl cAMP and 5 microM BayK 8644 induced a similar AHP increase. BayK 8644 occluded the effect of agonists. The results suggest that the action of dopamine is mediated via the recently described protein kinase A enhancement of L-type Ca2+ channels. The results partially explain the decrease in firing frequency induced by dopamine and a possible site of antagonism with cholinergic modulation. PMID- 8730805 TI - Altered dopamine transporter densities in alcohol-preferring vervet monkeys. AB - Altered dopamine (DA) transporter densities have been implicated in mechanisms of vulnerability and relapse in human alcoholics. The regional distribution and density of the DA transporter was studied in alcohol-preferring vervet monkeys to investigate baseline status and regulation of the DA transporter at different stages of chronic alcohol drinking. Combined ligand binding and in vitro autoradiography of the cocaine congener [125I]RTI-55 (beta-CIT) demonstrated a significant increase in DA transporter densities in abstinent alcohol-preferring monkeys over those in alcohol-avoiding monkeys. Chronic alcohol consumption down regulated DA transporter densities, and this effect was reversed by acute withdrawal. These results demonstrate that the DA transporter is regulated by alcohol exposure and suggest that increased DA transporter densities may be a phenotypic marker of alcohol preference in vulnerable monkeys. PMID- 8730806 TI - Cortical representation of self-paced finger movement. AB - We compared the cortical fields activated by simple, self-paced index-finger flexions with those activated during visually triggered movement and rest using PET. Of 12 fields detected during self-paced movement compared to rest, three were located in the classically defined motor areas: primary motor area (M1), premotor cortex (PM) and supplementary motor area (SMA). The latter extended into the cingulate motor area (CMA). Four corresponding clusters were also found when triggered movement was subtracted from self-paced movement. The change in regional cerebral blood flow (rCBF) was greater in SMA than in PM during self paced movement compared with either control. We conclude that repetitive, self paced index-finger flexions can activate SMA, PM and CMA, and that this movement activates SMA more strongly than PM. PMID- 8730807 TI - The cortical representation of object motion in man is interindividually variable. AB - Cortical areas processing visual motion have been well investigated in monkeys, but comparatively little is known about these areas in man. In order to define such cortical areas in the brains of individuals, the magnetic field was recorded while subjects were watching motion-defined static and moving objects. The magnetic response showed a transient component with a clear dipolar magnetic field followed by a sustained component which exhibited some variation in magnetic field structure over time. For the transient component, the single equivalent current dipoles superimposed upon magnetic resonance images for individual subjects were clearly localized outside the primary visual areas. In most cases the neural generator was found in the region of the temporo-parieto occipital junction of the lateral cortex. The results also suggest that the activated cortical area show interindividual variations in location. PMID- 8730808 TI - Retrograde neurotrophin-mediated control of neurone survival in the developing central nervous system. AB - Neuronal death in vertebrate development is widely believed to be regulated by retrograde survival signals from the axonal target territory, and these signals are assumed to be initiated by the binding of trophic molecules to the axon terminal. However, direct evidence for the retrograde transmission of such survival signals along developing axons is only available in the peripheral nervous system. We show here in a central projection, the isthmo-optic projection of chick embryos, that a neurotrophin, brain-derived neurotrophic factor, can indeed initiate retrograde survival signals from the target territory. A related molecule, neurotrophin-3, is ineffective. PMID- 8730809 TI - Presynaptic and Ca(2+)-independent PKC subspecies modulates NMDAR1 current. AB - We have studied the properties of the protein kinase C (PKC) subspecies that modulates the NMDA receptor (NMDAR1). The current through homomeric NMDAR1 expressed in Xenopus oocytes was increased by 200-500% by phorbol ester and also by activation of a metabotropic glutamate receptor (mGluR1) expressed in the same oocytes. This potentiation of the NMDAR1 current was not inhibited by the intracellular injection of EGTA. Intracellular injection of epsilon-PKC, a presynaptic PKC subspecies, potentiated the NMDAR1 current more efficiently that did the Ca(2+)-dependent gamma-PKC, a postsynaptic subspecies of the enzyme. Our findings suggested that the presynaptic NMDA receptor could be potentiated in a Ca(2+)-independent manner by the activation of presynaptic PKC subspecies. PMID- 8730810 TI - The stress of being restrained reduces brain damage after a hypoxic-ischaemic insult in the 7-day-old rat. AB - Experimental animals and patients are immobilized to allow continuous monitoring of physiological parameters. Restraint stress affects brain neurotransmitter levels and induces expression of immediate early genes. Whether immobilization stress affects outcomes in newborn animals which have suffered a hypoxic ischaemic insult is unknown. Twenty 7-day-old rats subjected to unilateral carotid ligation followed by 2 h hypoxia were randomly assigned to carry a rectal probe or to move freely. The 10 restrained animals showed 50% reduction in damage in all brain regions (p < 0.0.3). Plasma lactate levels, a marker of stress, were three times higher in animals carrying a rectal probe (p < 0.0.02) than those moving freely. We conclude that the stress of being restrained reduced damage after a hypoxic-ischaemic insult in the immature rat. PMID- 8730811 TI - Nerve growth factor stimulates production of neuropeptide Y in human lymphocytes. AB - The production of neuropeptide Y (NPY) in lymphocytes obtained from human tonsils was investigated using radioimmunoassay. While unstimulated lymphocytes did not produce detectable amounts of NPY, NPY synthesis was induced after cell activation. Our results show that the addition of nerve growth factor (NGF) to unstimulated lymphocytes has an effect similar to that of mitogens, both leading to production of NPY. The study of purified B and T cells confirmed that only activated cells are able to synthesize NPY. The stimulatory effect of NGF on NPY production is not a common characteristic of all lymphocytes: only unstimulated T cells respond to NGF by synthesizing NPY. No such effects is seen in purified B cells. PMID- 8730812 TI - Neuronal defects in genotyped dominant megacolon (Dom) mouse embryos, a model for Hirschsprung disease. AB - Dominant megacolon (Dom) is one of four mutations in the mouse which can produce a phenotype similar to Hirschsprung disease in man. Here, we report that it is possible to take advantage of two microsatellite markers to genotype Dom embryos and to study enteric neuronal development in Dom embryos using whole-mount immunohistochemistry. Dom embryos present a variable defect in the ileo-caecal region, as do embryos of other murine models of Hirschsprung disease. PMID- 8730813 TI - Nuclear accumulation of p53 protein following kainic acid-induced seizures. AB - The p53 tumor suppressor gene has been implicated in apoptotic cell death. The present study was conducted to investigate whether expression of p53 protein is increased in association with kainic acid-induced neuronal apoptosis. Adult male Sprague-Dawley rats were treated systemically with the glutamate analog kainic acid, and sacrificed either 4 or 30 h after the onset of seizure activity. Immunohistochemistry was performed on paraffin-embedded sections using an anti p53 polyclonal antibody. At both time points, increased p53 immuno-reactivity was observed predominantly in the nucleus of apoptotic neurons. These findings lend additional support to the hypothesis that p53 is a marker of neuronal apoptosis in the CNS, and suggest that nuclear accumulation of p53 protein may be an important mediator of neuronal death. PMID- 8730814 TI - An adenovirus encoding CuZnSOD protects cultured striatal neurones against glutamate toxicity. AB - Superoxide dismutase (SOD), a key enzyme in the detoxification of free radicals, catalyses the dismutation of superoxide O2.- to oxygen and hydrogen peroxide (H2O2). It is therefore a promising candidate for gene transfer therapy of neurological diseases in which free radicals are thought to be involved. We have constructed a recombinant adenoviral vector containing the human copper-zinc SOD cDNA. Using this vector we were able to drive the production of an active human copper-zinc SOD protein (hCuZnSOD) in various cell lines and primary cultures. Infection of striatal cells with a recombinant adenovirus expressing hCuZnSOD protected these cells from glutamate-induced cell death. PMID- 8730815 TI - Ageing potentiates anaesthetic-induced synaptic depression in hippocampal slices. AB - The effects of the volatile anaesthetic isoflurane on dendritic field excitatory postsynaptic potentials (fEPSP) were compared in hippocampal slices taken from young mature and old Fisher 344 rats. Application of isoflurane (1% v/v) to young brain slices produced minimal effects on the recorded fEPSPs. On the contrary, the same anaesthetic concentration depressed field responses obtained from old hippocampal slices by 42 +/- 6.8% compared with baseline values. Such increased sensitivity to anaesthetic action in the old slices was consistently observed with administration of higher isoflurane concentrations. The presynaptic afferent volley was unaffected by application of low or high anaesthetic concentration, suggesting that age-induced changes in nerve fibre conduction and probably nerve ending excitability are not involved in the increased vulnerability of old synapses to anaesthetic action. Other synaptic sites are probably involved in the mechanisms of age-dependent potentiation of anaesthetic suppression of synaptic transmission. PMID- 8730816 TI - Species and sex differences in hippocampus size in parasitic and non-parasitic cowbirds. AB - To test the hypothesis that selection for spatial abilities which require birds to locate and to return accurately to host nests has produced an enlarged hippocampus in brood parasites, three species of cowbird were compared. In shiny cowbirds, females search for host nests without the assistance of the male; in screaming cowbirds, males and females inspect hosts' nests together; in bay winged cowbirds, neither sex searches because this species is not a brood parasite. As predicted, the two parasitic species had a relatively larger hippocampus than the non-parasitic species. There were no sex differences in relative hippocampus size in screaming or bay-winged cowbirds, but female shiny cowbirds had a larger hippocampus than the male. PMID- 8730817 TI - Decreased density of I2-imidazoline receptors in the postmortem brain of heroin addicts. AB - The aim of this study was to quantitate imidazoline receptors in postmortem brains of heroin addicts who died of an opiate overdose. The density of I2 imidazoline receptors ([3H]idazoxan binding in the presence of adrenaline) and the immunoreactivity of the related 29/30 kDa imidazoline receptor protein were decreased (39% and 28%, respectively) in the frontal cortex. The density of brain I2-imidazoline receptors was also decreased in heroin-dependent rats (27%). This novel finding indicates that opiate addiction induces down-regulation of I2 imidazoline receptors in astrocytes, and presumably down-regulation of the functions associated with these receptors. PMID- 8730818 TI - Involvement of the presynaptic dopamine D2 receptor in the depression of spinal reflex by apomorphine. AB - The relative roles of D1 and D2 dopamine (DA) receptors in mediating apomorphine (APO)-induced changes in the spinal reflex was investigated. Low doses of APO, a DA receptor agonist (0.2 mg kg-1, i.v.), depressed the monosynaptic mass reflex (MMR) in spinalized rats. Pretreatment with the D2-specific antagonist, spiperone, 10 min before APO prevented the APO-induced MMR depression. Pretreatment with the D1 antagonist SCH 23390 failed to prevent the APO-induced depression. Interestingly, SCH 23390 pretreatment preferentially antagonized the depression induced by a high dose of APO (3 mg kg-1, i.v.). Pretreatment with SKF 38393, a selective D1 agonist, completely prevented the APO-induced MMR depression. These results suggest that inhibition of spinal transmission by low dose of APO may be mediated through its action on presynaptic D2 receptors and that D1 and D2 receptors are functionally coupled at the spinal level in modulating the spinal motor output. PMID- 8730819 TI - Cellular organization of lateral hypothalamic efferents to the central amygdaloid nucleus of the rat. AB - The present study aimed to describe the localization, organization and density of lateral hypothalamic neurones projecting to the central amygdaloid nucleus (CeA) using WGA-apoHRP-Au as a retrograde tracer. In the lateral hypothalamus (LH) a cluster of labelled neurones was located in the middle and posterior parts of this region. This retrograde labelling is organized into perifornical and juxtacapsular subpopulations. These results provide a neuroanatomical basis for the concept that these lateral hypothalamic neurones modulate projections from CeA to pons and brainstem autonomic and gustatory centres which integrate responses related to fundamental and adaptive behaviours, and could be related to self-stimulation behaviour elicited from the CeA. PMID- 8730820 TI - LTD and LTP induced by transcranial magnetic stimulation in auditory cortex. AB - Using a system capable of relatively localized and rapidrate transcranial magnetic stimulation (rTMS), evoked trains of complex spikes were studied in rodent auditory cortex. Low rate rTMS from 1 to 10Hz produced a frequency dependent increase in spike rate. Iterations of rTMS resulted in long-term potentiation (LTP)-like, and more durable long-term depression (LTD)-like changes in evoked spike rate. These observations generate new perspectives for studying mechanisms of learning and memory non-invasively as well as introducing some caveats for use of rTMS in humans. PMID- 8730821 TI - Mechanisms of allodynia: interactions between sensitive mechanoreceptors and nociceptors. AB - We examined whether stimulation of sensitive mechanoreceptors from an area of allodynia evokes nociceptor activity expressed as axon reflexes. Experiments were conducted on human volunteers. Cutaneous blood flow was measured with a laser Doppler flowmeter. Allodynia was induced with mustard oil (25-100%) or by intradermal injections of capsaicin (25-50 micrograms) in the skin of the forearm or the hand. Tactile stimulation of normal skin or outside zones of allodynia did not evoke axon reflexes. The same stimulation in areas of allodynia evoked pain as well as axon reflexes. Cooling the area of primary hyperalgesia or blocking the A fibres in the nerve that innervated the allodynia area abolished the allodynia and the axon reflex. These results demonstrate central interactions between sensitive mechanoreceptors and nociceptors concomitant with the development of allodynia. PMID- 8730822 TI - Modulation of IL-1 beta gene expression in the rat CNS during sleep deprivation. AB - We hypothesize that sleep homeostasis involves, at least in part, the immune system modulator interleukin-1 beta (IL-1 beta). Using the reverse transcription polymerase chain reaction, IL-1 beta mRNA levels in the rat CNS were evaluated after a period of sleep deprivation. In addition, IL-1 beta gene expression was analyzed before the projected onset of activity and rest phase in free-running animals. No changes in IL-1 beta mRNA were observed in the circadian cycle, but 24 h of sleep deprivation resulted in a 2-fold increase in the level of IL-1 beta mRNA in the hypothalamus and in the brain stem compared with controls (p < 0.0002 and (p < 0.0001 respectively). The alteration in IL-1 beta mRNA levels following sleep deprivation supports the hypothesis that modulation of IL-1 beta gene expression is involved in the sleep homeostatic process. PMID- 8730823 TI - Polymorphism in AACT gene may lower age of onset of Alzheimer's disease. AB - The ApoE-epsilon 4 allele is a predisposing factor for late onset Alzheimer's disease (AD), however it is neither necessary nor sufficient to cause the disease. A candidate for explaining part of the remaining genetic component is alpha 1-antichymotrypsin (AACT). In a case-control study we genotyped a polymorphism within the AACT gene to test for association with the disease. No allele of this gene showed an increased incidence among the population with AD compared with controls, even when taking ApoE genotype into account. This contrasts with the results of a recently published report. The mean age of onset was apparently lowered by the presence of the AACT AA genotype among ApoE-epsilon 4 bearers. If AACT genotype has an effect on risk for AD it may be predominantly amongst individuals with early onset AD. PMID- 8730824 TI - Electrophysiological properties of Hensen's cells investigated in situ. AB - Tight-seal whole-cell patch clamp recordings were obtained in situ from supporting Hensen's cells within the intact organ of Corti of the adult guinea pig. In normal phosphate buffer solution we estimated 20-50 cells to be coupled by gap junctions to the cell under the patch pipette. In the presence of 1 mM octanol, an uncoupling agent, it was possible to identify an outward current which activated upon depolarization above -20 mV and approached saturation above 70 mV. An inward current was seen with hyperpolarizations below -80 mV. These are broadly similar to the currents of Hensen's cells in vitro. Measured differences of the underlying conductance indicate that the currents are sensitive to the procedure used to isolate cells. PMID- 8730825 TI - Non-synaptic and dendritic exocytosis from dense-cored vesicles in the suprachiasmatic nucleus. AB - The sites of exocytosis by dense-cored vesicles (DCVs) from neurones in the rat suprachiasmatic nucleus (SCN) were studied at the ultrastructural level. The tannic acid procedure, which stabilizes extruded proteinaceous substances, was used to demonstrate exocytosis of DCV cores. Fresh brian slices containing the SCN were incubated in media containing high levels of potassium (56 mM) or glutamate (10 mM) in the presence of tannic acid. Long-term slice explant cultures of the SCN were similarly treated. Exocytosis from DCVs occurred from axonal terminals, from dendrites, and occasionally from somata. The sites of DCV exocytosis were generally nonsynaptic or para-synaptic, including release immediately adjacent to axo-spinous synaptic densities. These observations show that, in the suprachiasmatic nucleus, neuroactive substances contained in DCVs do not necessarily function at synaptic contract zones, but could also act as neuromodulators at non-synaptic sites. PMID- 8730826 TI - Enhancement of NMDA receptor maturation by BDNF in cultured mouse cerebellar granule cells. AB - The N-methyl-D-aspartate (NMDA) subtype of glutamate receptor, which is essential during organogenesis for neuronal differentiation and formation of synaptic connections in the developing brain, changes its receptor properties during development by differential expression of multiple subunits. Using a combined electrophysiological and pharmacological approach on primary cultures of mouse cerebellar neurones, we investigated the evolution of the NMDA receptor and the potential effect of the neurotrophin BDNF on its expression. We showed that 1) the current density of NMDA responses increased with time of culture; 2) epsilon 1 subunit expression increased with time in culture relative to epsilon 2 subunit expression; and 3) the time course of the increase in NMDA responses was accelerated by about 2 days in the presence of BDNF. PMID- 8730827 TI - Visual evoked potentials are affected by trunk rotation in neglect patients. AB - Steady-state visual evoked potentials (VEPs) were recorded in four patients with unilateral visuo-spatial neglect, stimulating either the left or the right hemifield. In the standard condition (head and body oriented straight ahead towards the stimulus) the left hemifield VEP was delayed. When the body was turned to the left, however, the two hemifield latencies were comparable. These results were confirmed with the transient VEP technique. No effect of trunk rotation was observed in a group of patients with left brain damage and without neglect. The results imply that the sensory afferents from neck muscles might restore the altered occipital activity and suggest that the same conditions which modulate neglect modulate VEPs latencies. PMID- 8730828 TI - Morphological alterations of the visual system in white zebra finches. AB - Visual system anomalies in albino mammals are generally caused by a lack of retinal pigment and misrouting of retinofugal optic fibres. This study shows that the visual system of white zebra finches differs morphologically from that of normally coloured (wild type) birds, although eye pigmentation and retinofugal projection are normal. In white zebra finches, the recrossing tectorotundal projection is enhanced and a new (reciprocal) connection is established between the n. rotundi of both hemispheres. The morphological alterations found in this study may contribute to the previously described enhanced processing of ipsilateral stimuli in white zebra finches. PMID- 8730829 TI - Loss of vessel wall viability in cerebral amyloid angiopathy. AB - Cerebral amyloid angiopathy (CAA) is a neuropathological feature of Alzeheimer's disease and an important cause of cerebral haemorrhage in the elderly. CAA is characterized by the deposition of Alzheimer amyloid beta protein (A beta) in cerebral and leptomeningeal vessel walls. In order to study the effect of cerebrovascular A beta deposits in vivo, living canine leptomeninges obtained from old dogs affected by CAA were analysed by confocal laser scanning microscopy after immunofluorescence staining for A beta and viability staining with fluorescein diacetate (FDA). Simultaneous detection of the two signals showed a segmental loss of leptomeningeal vessel wall viability at some site of A beta deposition. Many of the non-viable vessels segments were also dilated, suggesting that A beta-induced vascular cell death creates the loci minores resistentiae for the development of cerebral haemorrhage in CAA. PMID- 8730830 TI - Cholinergic-glutamatergic interactions in visual recognition memory of rhesus monkeys. AB - Administration of either a non-competitive N-methyl-D-aspartate (NMDA) receptor antagonist, MK-801 (dizocilpine), or a muscarinic antagonist, scopolamine, produces dose-related impairments in performance of a visual recognition memory task, delayed non-matching to sample (DNMS) with lists of 20 symbols, in rhesus monkeys. In the present study, low doses of these two drugs, which were ineffective when given alone, significantly impaired performance when given in combination. Moreover, the effect was greater than additive, indicating a synergistic interaction. These results suggest that interactions between cholinergic and glutamatergic systems play an important role in regulation of visual recognition memory. PMID- 8730831 TI - Nitric oxide-stimulated increase in intracellular cGMP modulates gap junction coupling in rat neocortex. AB - In the present study we demonstrate that gap junction coupling between developing layer II/III pyramidal cells in rat sensorimotor cortex is strongly modified by the nitric oxide (NO)/cyclic guanosine monophosphate (cGMP) system. Dye coupling was revealed by intracellular injection of the gap junction-permeable tracer neurobiotin into single neurones. Following incubation of slices with sodium nitroprusside, a source of NO, the size of dye-coupled cell clusters was significantly reduced. In many cases, 2-3 cells remained strongly dye-coupled. These effects were blocked by intracellular injection of the guanylyl cyclase inhibitor cystamine and mimicked by both application of the membrane-permeant cGMP analogue 8-Br-cGMP and intracellular injection of cGMP. cGMP injection also induced a 60% increase in neuronal input resistance. These results indicate that NO modulates gap junction coupling in the developing neocortex via stimulation of guanylyl cyclase. PMID- 8730832 TI - PTX-sensitive regulation of neurite outgrowth by the dopamine D3 receptor. AB - Neurotransmitter receptors are known to have direct roles in the modulation of neuronal morphogenesis. Previous work showed that clonal mesencephalic MN9D cells individually transfected with D2-like dopamine receptors show increased neurite outgrowth following long-term exposure to the D2-like receptor agonist quinpirole. In the current study, brief stimulation of D3 receptor-expressing cells also elicited increased neurite outgrowth, which could be mimicked by the Gi/G(o) protein activator mastoparan. Pretreatment with the Gi/G(o) protein inhibitor pertussis-toxin blocked the quinpirole- and mastoparan-mediated increases in outgrowth. These results suggest that dopamine D3 receptor stimulation has an immediate, G-protein-mediated role in neuronal morphogenesis. PMID- 8730833 TI - Prolonged enhancement of REM sleep produced by carbachol microinjection into the amygdala. AB - The effect on sleep organization of carbachol microinjected into different amygdaloid nuclei was analysed in 12 cats. Single carbachol doses of 8 micrograms in 0.50 microliter saline were delivered unilaterally or bilaterally into the central, basal, lateral or basolateral amygdaloid nucleus. Carbachol administration into the central nucleus induced a prolonged (5 days) enhancement of both REM sleep and its preceeding slow wave sleep episodes with PGO waves (sommeil phasique a ondes lentes, SPHOL), which was more pronounced following bilateral than unilateral carbachol administration. However, neither SPHOL nor REM sleep changes were produced by administration of carbachol into the other amygdaloid nuclei. We conclude that cholinergic activation of the central amygdaloid nucleus produces a long-term facilitation of REM sleep occurrence. PMID- 8730834 TI - Ultrasonic vocalization (22-28 kHz) in a model of chronic pain, the arthritic rat: effects of analgesic drugs. AB - Adjuvant-induced arthritis in rats is considered as a chronic pain model. In a search for a relevant behavioural marker of the chronic pain state, we studied social interactions between an arthritic rat and a healthy partner in a neutral cage. During the test, arthritic rats emitted ultrasonic 22-28 kHz vocalizations (USV), whereas control rats did not, and showed less exploration and more immobility, but did not differ from controls in terms of social behaviours (social investigation, allogrooming). Aspirin (200 mg kg < > (en)1) or morphine (3 mg kg < > (en)1) injected intraperitoneally before the test significantly decreased USV without affecting the animals' behaviour. These results suggest that USV could be associated with affect related to aversive or painful stimuli and may constitute a behavioural marker of chronic pain in arthritic rats. PMID- 8730835 TI - Hippocampal cGMP and cAMP are differentially involved in memory processing of inhibitory avoidance learning. AB - Cyclic GMP (cGMP) and cyclic AMP (cAMP) have been proposed to participate in the early and late stages of long-term potentiation (LTP), respectively. Here we report on the effect of post-training intrahippocampal infusion of membrane permeable analogues of these cyclic nucleotides on the consolidation of inhibitory avoidance learning in rats and on the effect of this task on hippocampal cGMP and cAMP levels. Bilateral intrahippocampal microinjection of 8 Br-cGMP (1.25 micrograms per side) enhanced retention test performance when given immediately (0 min), but not when given 180 min, after training. In marked contrast, intrahippocampal infusion of the same dose of 8 Br-cAMP facilitated memory consolidation when given 180 min, but not 0 min, after training. Rats submitted to an inhibitory avoidance task showed a significant increase in the amount of cGMP in the hippocampus at 0 and 30 min after training, and in the amount of cAMP 30 and 180 min after training. Taken together, these results indicate that cGMP-regulated processes in the hippocampus play an important role in the early stages of memory consolidation and that cAMP signalling pathways are involved in the late post-training memory processing of inhibitory avoidance learning. PMID- 8730836 TI - On orienting the hand to reach and grasp an object. AB - Subjects were required to reach and grasp a parallelepiped, the position, orientation and size of which were varied. The kinematics of reaching and grasping movements was studied in full vision and in no vision conditions. Both direction and movement amplitude of reaching were affected by object orientation. Conversely, both the time course of finger axis orientation and the angular displacement of the hand at wrist were influenced by object position. These results were not modified by the absence of visual control. Finger aperture during grasping was affected by both object size and orientation. This latter result was not due to a distorted size perception, as shown by a control matching experiment. Taken together, the results of the present study suggest the integration between distal and proximal components during reaching and grasping. PMID- 8730837 TI - Antisense oligodeoxynucleotide treatment to the brain cannabinoid receptor inhibits antinociception. AB - Studies using agonists from at least three major cannabinoid ligand groups suggest the mediation of several distinct effects (e.g. psychotropic, analgesia, and antiemetic) by the recently cloned CB1 cannabinoid receptor. However, other studies suggest the presence of multiple cannabinoid receptors and at least one other receptor (CB2) has been cloned. The present investigation was undertaken to determine whether one of the potential therapeutic actions of cannabinoids (i.e. antinociception) is mediated by the CB1 receptor using the antisense oligodeoxynucleotide 'knock-down' approach. Synthetic oligodeoxynucleotides complementary to the 5' end of the coding region of the mouse CB1 receptor mRNA were administered to mice by the intracerebro-ventricular (i.c.v.) route twice daily for 3 days. Mismatch oligodeoxynucleotides of similar sequence, but containing six mismatched positions out of the 18 nucleotides within the oligodeoxynucleotide were administered to other mice. Treatment with antisense oligodeoxynucleotides, but not mismatched oligodeoxynucleotides, greatly inhibited the antinociceptive response of the cannabinoid agonist CP-55,940. Untreated mice and those treated with mismatched oligodeoxynucleotides showed similar, full response antinociception after CP-55,940 administration. The data provides strong evidence that the CB1 receptor-ligand interaction is essential for the antinociceptive effect. PMID- 8730838 TI - Melatonin-dopamine interaction in the striatal projection area of sensorimotor cortex in the rat. AB - The excitatory response to motor cortex stimulation of 201 striatal neurones was recorded electrophysiologically to test the effects of melatonin (aMT) and/or D1 and D2 antagonists. Iontophoresis of aMT attenuated the excitatory response in 68.5% of neurones, with a latency of 2-4 min and enhanced the excitatory response in 11.9% of the neurones; 19.6% showed no change in response. Iontophoresis of sulpiride (D2 antagonist) produced an immediate increase in the excitatory response in 62.8% of neurones, an attenuation in 2.3% and no change in the response of 34.9%. The ejection of sulpiride counteracted the aMT-dependent inhibition of the excitatory response of striatal neurones. SCH-23390 (D1 antagonist) iontophoresis had no significant effect. The results show that the same striatal units may be driven by aMT and D2 receptors. However, the significant difference in the latency of the responses suggests that the effects of these two substances are mediated by different receptor/intracellular messengers. PMID- 8730839 TI - Expression of the neurofibromatosis 1 (NF1) gene during growth arrest. AB - The neurofibromatosis 1 (NF1) gene product, neurofibromin, is a tumor suppressor gene product capable of inhibiting the growth of cells in culture. If neurofibromin suppresses cell growth by arresting cells in G0 or G1, its expression might be regulated in a cell cycle-dependent fashion. In this study, we demonstrate that RAT-1A fibroblasts arrested in G0/G1 by serum starvation and then released to progress through the cell cycle do not demonstrate significant changes in NF1 expression. However, when arrested in G0/G1 by contact inhibition, NF1 expression in these cells is reversibly upregulated within 72 h, suggesting that NF1 expression is a late event associated with cell growth arrest which may contribute to the maintenance of the differentiated state. PMID- 8730840 TI - Nitric oxide regulates NMDA-induced dopamine release in rat striatum. AB - We investigated the effect of nitric oxide (NO) on N-methyl-D-aspartate (NMDA) induced changes in levels of dopamine (DA) and its metabolite in the striatum using in vivo microdialysis. Local administration of 1mM NMDA into the striatum significantly augmented DA release in the striatum. Simultaneous administration of -5mM N(G)-nitro-L-arginine methyl ester (-NAME), a NO synthase inhibitor, into the striatum significantly potentiated NMDA-induced DA release. This effect of L NAME was completely reversed in the presence of 50mM L-arginine (L-Arg). Administration of 1 mM NMDA significantly decreased the levels of dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA). This effect of NMDA was not affected by concurrent administration of L-NAME. This study provides in vivo evidence for the involvement of NO in NMDA-induced DA release. PMID- 8730841 TI - Liposome-mediated BDNF cDNA transfer in intact and injured rat brain. AB - We examined the temporal profile of the expression of brain-derived neurotrophic factor (BDNF) cDNA containing a viral promotor following the injection of liposome cDNA complexes into the intact and traumatically injured rat brain. In situ hybridization and PCR confirmed the presence of injected BDNF cDNA for at least 6 days after injection. A similar profile of BDNF cDNA was observed when it was injected following cortical impact injury. mRNA was also localized around the injection areas. These results suggest that liposome-mediated delivery of neurotrophin cDNA may be a practical gene transfer method for treating traumatic brain injury. PMID- 8730842 TI - Nitrooxy alkyl apovincaminate modulates GABA(A) receptor in rat neocortical neurones. AB - We investigated the effects of VA-045 ((+)-eburnamenine-14 carboxylic acid (2 nitroxyethyl) ester) on GABA(A) receptor-mediated Cl- currents in dissociated rat cerebral cortical neurones, using a nystatin-perforated patch recording configuration. At a holding potential of - 40mV, the external application of GABA evoked an inward Cl- current with an EC50 value of 5.6 x 10(-6)M. VA-045 increased the GABA response at GABA concentrations below 3.0 x 10(-6)M. The GABA response showed a time dependent decay consisting of fast and slow components and VA-045 significantly accelerated both components. The site of action for VA-045 was considered to be different from that for benzodiazepines or barbiturates. PMID- 8730843 TI - Selective behavioral alterations on addition of a 4'-phenyl group to cocaine. AB - We synthesized a cocaine analog in which a phenyl group was added at the para position of the benzene ring of cocaine. This substitution caused a modest reduction (four-fold compared with cocaine) in binding potency for the primate (Papio) dopamine transporter as judged by displacement of [3H]WIN 35,428 binding from caudate/putamen membranes. Behavioral effects of this structural modification in the mouse were complex and selective, comprising absence of stimulation of locomotor activity, enhanced inhibition of locomotion and reduced lethal potency. Convulsant potency was unaltered. Substituents at the 4'-position of cocaine are important in its actions. Simple changes in the chemical structure of this drug may produce complex and selective changes in its neurochemical and behavioral actions. PMID- 8730844 TI - A calbindin immunoreactive "deep pain' recipient thalamic nucleus in the rat. AB - The ventrolateral periaqueductal gray (vlPAG) is the only brain region known to receive convergent deep noxious inputs and to mediate the reactions characteristic of deep pain. Injections of biotinylated dextran into the vlPAG of the rat revealed a strong projection to a discrete, calbindin terminal immunoreactive region of the caudal ventromedial nucleus (VMc) of the thalamus. This nucleus appears homologous to the calbindin-positive, pain- and temperature specific-posterior ventromedial thalamic region of primates. We suggest that the vlPAG to VMc projection represents an important new route via which deep noxious inputs reach thalamus. As the rat is the species of choice in most experimental studies of pain, the functional-anatomical definition of this projection should further investigation of the thalamic representation of deep pain. PMID- 8730845 TI - Intrastriatal injection of GDNF attenuates the effects of 6-hydroxydopamine. AB - Our study was designed to determine whether intrastriatal administration of glial cell line-derived neurotrophic factor (GDNF) can attenuate the behavioral effects and injury to the mesostriatal dopaminergic system caused by 6-hydroxydopamine (6 OHDA). Four groups of rats received a series of four intrastriatal injections of vehicle or one of three doses of GDNF (0.1, 1 or 10 micrograms per injection) on days 1,3,5 and 8. On day 4 the animals received a single, intrastriatal injection of 25 micrograms 6-OHDA. Treatment with GDNF significantly reduced the development of amphetamine-induced rotation, and the dose of 1 microgram per injection appeared to be the most effective. The group treated with this dose had significantly greater preservation of tyrosine hydroxylase-immunoreactive (TH-IR) fibers adjacent to the injection site in the striatum and significantly greater preservation of Nissl-stained and TH-IR neurons in the substantia nigra pars compacta (SNpc). PMID- 8730846 TI - The movement of IGF-I into the brain parenchyma after hypoxic-ischaemic injury. AB - The movement of peptides from CSF into the parenchyma is thought to be slow and diffusion limited. However IGF-I can reduce neuronal loss at distal sites when given centrally 2 h after hypoxic-ischaemic (HI) injury. The present study determined the distribution of [3H]IGF-I given into the lateral ventricle after unilateral HI injury in adult rats. Radioactivity in the injured cortex peaked immediately after administration then rapidly declined. Autoradiography demonstrated radioactivity in the perivascular spaces and in the corpus callosum and external capsule of the injured hemisphere. HPLC and radioimmunoassay confirmed a rise in intracerebral IGF-I levels (from 159 +/- 9 to 401 +/- 88 ng g 1). These data suggest that injury can enhance the movement of IGF-I into the cerebrum via the white matter tracts and perivascular spaces. PMID- 8730847 TI - Brain ischaemia transiently activates Ca2+/calmodulin-independent protein kinase II. AB - The activity of Ca2+/calmodulin-dependent protein kinase II (CaM-KII) during short-term global ischaemia was investigated in the gerbil brain hippocampus and cortex. Ischaemia of 0.5 min duration significantly stimulated Ca(2+)-independent 'autonomous' activity, indicating activation of the first step of intramolecular enzyme phosphorylation just after ischaemia has developed. Prolongation of the ischaemic period up to 5 min inhibited both Ca(2+)-dependent and, to a lesser extent, Ca(2+)-independent activities of CaM-KII. These last events coincide with an extensive translocation of CaM-KII protein from the soluble to the membranous fraction. In effect, in spite of inhibition of total CaM-KII activity, its Ca(2+) independent, persistently active component can still remain more abundant at specific membrane regions. PMID- 8730848 TI - Inhibition of tumor necrosis factor attenuates physiological sleep in rabbits. AB - The involvement of tumor necrosis factor (TNF) in sleep regulation was investigated by blocking TNF using a synthetic TNF receptor fragment (TNFRF) in rabbits. Intracerebroventricular injection of 50 micrograms TNFRF decreased spontaneous non-rapid-eye-movement (NREM) sleep across the 21 h recording period. After 6 h of sleep deprivation (SD), both duration of NREM sleep and electroencephalographic (EEG) slow wave activity during NREM sleep were enhanced. TNFRF suppressed SD-enhanced NREM sleep and EEG slow wave activity. SD per se did not affect rapid-eye-movement (REM) sleep, but TNFRF treatment increased REM sleep after SD. These results are consistent with the hypothesis that TNF is involved in the regulation of physiological NREM sleep. PMID- 8730849 TI - Ipsilateral and bilateral receptive fields in rat primary somatosensory cortex. AB - The activity of single neurones evoked by electrical stimulation of the paws was recorded from the central forepaw and hindpaw receptive fields of the primary somatosensory cortex (Sm1) in anaesthetized rats. In the forepaw receptive area (FRA), 44% of neurones showed pyramidal responses to stimulation of their central receptive fields (CRF) compared with 13% of neurones in the hindpaw receptive area (HRA). The neurones were located primarily in layers IV and V. A number of neurones in superficial layers of the hindpaw region, but not in the forepaw region, had either bilateral or exclusively ipsilateral receptive fields. It was concluded that neurones with bilateral hindpaw receptive fields form a separate population of neurones which are organized somatopically. PMID- 8730850 TI - 'Prepulse inhibition' facilitates a liberal response bias in an auditory discrimination task. AB - This study examined 'prepulse inhibition' in the context of an auditory two-tone discrimination task performed by 15 healthy subjects. In order to distinguish between masking or excitatory information processes, weak acoustic pulses immediately preceded or succeeded a target or non-target tone in the discrimination. Button-press performance was compared with response in a no-pulse condition. Response bias (beta) became more liberal in the 100 ms prepulse and 200 ms postpulse conditions. Beta correlated with P3b amplitude measures of the event-related potential. The weak pulses increased temporarily the cortical excitability, as measured by the decreased amplitude of the P3b component and thus facilitated a more liberal response bias. PMID- 8730851 TI - Association of phosphotyrosine with rapsyn expression in Xenopus embryonic cells. AB - The postsynaptic membrane of the neuromuscular junction is highly enriched in rapsyn, which is thought to interact directly with nicotinic acetylcholine receptors (AChR) and anchor them at the synapse. We expressed rapsyn with or without AChRs in Xenopus embryos by mRNA injection. Co-expression of AChR and rapsyn caused the clustering of these two proteins in cultured cells isolated from the injected embryos. When rapsyn was expressed alone, it also became clustered at the substratum-facing membrane in cultured cells and at cell-cell contacts in whole mount embryos. No clusters were observed in cells that expressed AChRs alone. In rapsyn-expressing cells, proteins that are tyrosine phosphorylated as shown by anti-phosphotyrosine antibody labeling were concentrated at rapsyn clusters. Rapsyn itself does not appear to be a substrate for tyrosine kinase. This suggests that other phosphotyrosine-containing proteins are co-clustered with rapsyn in these cells. PMID- 8730852 TI - Neuroprotection against CA1 injury with metalloporphyrins. AB - The hippocampal slice was used to examine neuroprotection with metalloporphyrins, a class of drug which inhibits heme oxygenase and which has been found to be effective in the treatment of neonatal hyperbilirubinemia. Tin-protoporphyrin given during hypoxia significantly improved recovery of CA1 antidromic PS to a mean of 82 +/- 2% of initial amplitude, while unmedicated slices regained only 6 +/- 3% of initial amplitude. Tin-protoporphyrin also protected against fluid percussion injury with an EC50 of 10 microM when given after trauma. This protection extended to induction of long-term potentiation. Tin-mesoporphyrin and zinc-protoporphyrin protected against trauma with EC50's of 4 and 32 microM. Treatment with Sn-PP also protected against exposure to hydrogen peroxide, but not NMDA, AMPA, glycine or nitric oxide. These findings indicate that metalloporphyrins protect against CA1 neuronal injury through direct neural effects. PMID- 8730853 TI - HB-GAM is a cytokine present in Alzheimer's and Down's syndrome lesions. AB - The distribution of heparin binding growth associated molecule (HB-GAM) in the cerebral amyloidoses of Alzheimer's disease (AD) and Down's syndrome (DS), conditions characterized by the deposition of amyloid beta (A beta), was investigated immunohistochemically. Antibodies to HB-GAM, a cytokine which plays an important role in brain development and maturation, showed strong immunoreactivity with senile plaques in both AD and DS. Anti-HB-GAM reacted with pre-amyloid lesions, but only when markers of dystrophic neurites were present. The presence of HB-GAM in AD brains, but not in control brains, was confirmed by Western blotting. We suggest that the presence of HB-GAM in A beta lesions is a marker of neuronal injury. PMID- 8730854 TI - Effects of colour and lexical relevance on irrelevant stimuli: event-related potential results. AB - Event-related potentials (ERPs) were recorded in a warning stimulus-imperative stimulus (S1-S2) situation where S1 was either a coloured bar followed by a letter string (S2), or vice versa. When S1 was a coloured bar, reaction time (RT) response was required only to words which were preceded by a bar of a particular colour. When S1 was a letter string, response was required only to bars in a particular colour which were preceded by a word. Contingent negative variation (CNV) appeared only after those S1s which could be followed by a target stimulus. However, bars in the colour of the target elicited attention-related ERP components even if these stimuli were preceded by non-words, i.e. when the subjects could be certain in advance of the appearance of a non-target stimulus. PMID- 8730855 TI - Developmental regulation of voltage-activated Na+ and Ca2+ currents in rat retinal ganglion cells. AB - The developmental regulation of voltage-gated Na+ and Ca2+ current expression was investigated in rat retinal ganglion cells (RGCs) using whole cell patch clamp recordings. Experiments were performed on retinal whole mounts and slices from embryonic day 14 (E14), the developmental stage where RGCs start to differentiate from their precursor cells, to postnatal day 25 (P25) where the retina is fully differentiated and the animals have opened their eyes. No voltage-activated ion currents could be detected earlier than E17. From E17/18 on, small voltage-gated Na+ and Ca2+ currents could be measured which increased remarkably in amplitude until P16. Analysis of current kinetics and application of specific calcium channel antagonists revealed an alteration in the contribution of different Ca2+ current components to the voltage-activated whole cell Ca2+ current in RGCs during development. PMID- 8730856 TI - NT-4/5 protects against adrenalectomy-induced apoptosis of rat hippocampal granule cells. AB - Adrenalectomy (ADX) in rats has been shown to induce apoptosis of hippocampal granule cells. We tested whether neurotrophins are able to protect hippocampal neurons in this neurodegeneration model. Acid fucshsin stain was used to identify pyknotic cells in ADX rats treated for 4 days with NT-3, NT-4-5 or cytochrome-C, as a control protein. Cytochrome-C injections slightly decreased cell death on the ipsilateral side. NT-3 did not further promote this effect. Significantly less cell death was observed bilaterally in hippocampus treated with NT-4/5. TUNEL end labeling also confirmed the results. Our results demonstrated that NT 4/5, but not NT-3, promotes hippocampal neuron survival in adrenalectomized rats. They further show that injections of a control solution can induce a local protective effect. PMID- 8730857 TI - Aspartyl-phosphate phosphatases deactivate the response regulator components of the sporulation signal transduction system in Bacillus subtilis. AB - Bacteria use two-component signal transduction systems to sense and respond to their environment. A sensor kinase and a response-regulator transcription factor work in concert by phosphorylation/dephosphorylation through kinase and phosphatase activities to maintain a level of phosphorylated response regulator commensurate with the level of signal input. Signal input can be accommodated through stimulation of the kinase activity or the phosphatase activity of the two component system. With some notable exceptions, the sensor kinases recognize a single stimulatory ligand. A new dimension in the regulation of two-component signal transduction systems was discovered in the Rap phosphatases which dephosphorylate the SpoOF response-regulator of Bacillus subtilis independent of the sensor kinases. This family of phosphatases is encoded by at least six chromosomal genes. Although not all of the phosphatases of the family have activity on phosphorylated SpoOF, the two best-characterized members, RapA and RapB, prevent sporulation by dephosphorylating this response regulator component of the phosphorelay. Phosphatase activity of RapA is regulated by a gene, phrA, in the same transcriptional unit, that encodes a peptide secreted from the cell which may serve as a quorum sensor. Most of the Rap phosphatase operons have a gene coding for a protein with some similarity to PhrA in their transcription units, but it is uncertain whether all of these play a role in regulation. The Rap phosphatases are postulated to be a mechanism for allowing signals other than those that affect the sensor kinases to regulate the signal transduction pathway. They may have been recruited to help regulate sporulation because the multiple signals regulating this process may outstrip the recognition capacity of the kinases. PMID- 8730858 TI - Stationary phase in Saccharomyces cerevisiae. AB - Like other microorganisms, the yeast Saccharomyces cerevisiae responds to starvation by arresting growth and entering stationary phase. Because most microorganisms exist under conditions of nutrient limitation, the ability to tolerate starvation is critical for survival. Molecular analyses have identified changes in transcription, translation, and protein modification in stationary phase cells. At the level of translation, the pattern of newly synthesized proteins in stationary-phase cells is surprisingly similar to the pattern of proteins synthesized during exponential growth. When limited for different nutrients, yeast strains may not enter stationary phase but opt for pathways such as pseudohyphal growth. If nutrient limitation continues, the end-point is likely to be a stationary-phase cell. Based on the results of recent studies, we propose a model for entry into stationary phase in which G(o) arrest is separable from acquisition of the ability to survive long periods of time without added nutrients. PMID- 8730859 TI - The SMR family: a novel family of multidrug efflux proteins involved with the efflux of lipophilic drugs. AB - The sequenced members of a novel family of small, hydrophobic, bacterial multidrug-resistance efflux proteins, which we have designated the small multidrug resistance (SMR) protein family, are identified and analysed. Two distinct clusters of proteins were identified within this family: (i) small multidrug efflux systems; and (ii) Sug proteins, potentially involved in the suppression of groEL mutations. Hydropathy and residue distribution analyses of this family suggest a structural model in which the polypeptide chain spans the membrane four times as mildly amphipathic alpha-helices. The roles of specific residues, a possible mechanistic model of drug efflux, and the primary physiological role(s) of the SMR proteins are discussed. PMID- 8730860 TI - Regulatory noise in prokaryotic promoters: how bacteria learn to respond to novel environmental signals. AB - Various features of the regulation of pathways for biodegradation of recalcitrant compounds by Pseudomonas provide insights into the mechanisms by which operons evolve to acquire conditionally active promoters that permit the corresponding genes to be transcribed only when required. The "regulatory noise hypothesis' proposes that transcriptional control systems develop responsiveness to new signals due to the leakiness and lack of specificity of preexisting promoters and regulators. When needed, these may become more specific through suppression of undesirable signals and further fine-tuning of the recruited proteins to interact with distinct chemicals. This hypothesis is supported by the sophisticated regulation of sigma 54-dependent promoters of the TOL (toluene biodegradation) operons, which can be activated to various degrees by heterologous proteins. Such "illegitimate' activation is suppressed by bent DNA structures, either static or protein induced, between promoter core elements. Therefore, not only the regulators but also the DNA sequences participate in the process that gives rise to novel specificities. PMID- 8730861 TI - Extracellular addition of a domain of HIV-1 Vpr containing the amino acid sequence motif H(S/F)RIG causes cell membrane permeabilization and death. AB - Vpr is a virion-associated protein of human immunodeficiency virus type 1 (HIV-1) whose function in acquired immune deficiency syndrome (AIDS) has been uncertain. We previously employed yeast as a model to examine the effects of Vpr on basic cellular functions; intracellular Vpr was shown to cause cell-growth arrest and structural defects, and these effects were caused by a region of Vpr containing the sequence HFRIGCRHSRIG. Here we show that peptides containing the H(S/F)RIG amino acid sequence motif cause death when added externally to a variety of yeast including Saccharomyces cerevisiae, Kluyveromyces lactis, Candida glabrata, Candida albicans and Schizosaccharomyces pombe. Such peptides rapidly entered the cell from the time of addition, resulting in cell death. Elevated levels of ions, particularly magnesium and calcium ions, abrogated the cytotoxic effect by preventing the Vpr peptides from entering the cells. Extracellular Vpr found in the serum, or breakdown products of extracellular Vpr, may have similar effects to the Vpr peptides described here and could explain the death of uninfected bystander cells during AIDS. PMID- 8730862 TI - Alteration of haem-attachment and signal-cleavage sites for Paracoccus denitrificans cytochrome C550 probes pathway of c-type cytochrome biogenesis in Escherichia coli. AB - Paracoccus denitrificans cytochrome C550 is expressed as a periplasmic holo protein in Escherichia coli; amino acid substitutions of cysteine residues in the haem-binding motif (Cys-X-X-Cys-His), either together or singly, prevented covalent attachment of haem but not polypeptide translocation into the periplasm. When the three alanine residues at positions -3 to -1 in the native signal cleavage site were deleted, or alanine at -1 was changed to glutamine, signal cleavage was at alternative sites (after only ten residues in the latter case), but haem attachment still occurred. When the same three alanines were changed to Asp-Glu-Asp, a membrane-associated apo product that had retained the complete signal sequence was detected. These and other results presented here indicate that (i) haem attachment is not required for the apo-cytochrome C550 export to the periplasm; (ii) haem cannot attach to apo-cytochrome C550 when attached to the cytoplasmic membrane, suggesting that signal-sequence cleavage precedes periplasmic haem attachment, which can occur at as few as six residues from the mature N-terminus; and (iii) two cysteines are required for haem attachment, possibly because a disulphide bond is an intermediate. The gene for Saccharomyces cerevisiae mitochondrial iso-1-cytochrome c was expressed as a holo-protein in E. coli when fused with the signal sequence plus the first 10 residues of the mature cytochrome C550, indicating that the E. coli cellular apparatus for the c-type cytochrome biogenesis has a broad substrate specificity. PMID- 8730863 TI - Delta mu H+ dependency of in vitro protein translocation into Escherichia coli inner-membrane vesicles varies with the signal-sequence core-region composition. AB - Signal sequences frequently contain alpha-helix-destabilizing amino acids in the hydrophobic core. Nuclear magnetic resonance studies on the conformation of signal sequences in membrane mimetic environments revealed that these residues cause a break in the alpha-helix. In the precursor of the Escherichia coli outer membrane protein PhoE (pre-PhoE), a glycine residue at position -10 (Gly -10) is thought to be responsible for the break in the alpha-helix. We investigated the role of this glycine residue in the translocation process by employing site directed mutagenesis. SDS-PAGE analysis showed drastic variations in the electrophoretic mobilities of the mutant precursor proteins, suggesting an important role of the glycine residue in determining the conformation of the signal sequence. In vivo, no drastic differences in the translocation kinetics were observed as compared with wild-type PhoE, except when a charged residue (Arg) was substituted for Gly -10. However, the in vitro translocation of all mutant proteins into inverted inner-membrane vesicles was affected. Two classes of precursors could be distinguished. Translocation of one class of mutant proteins (Ala, Cys and Leu for Gly -10) was almost independent of the presence of a delta mu H+, whereas translocation of the other class of precursors (wild type or Ser) was strongly decreased in the absence of the delta mu H+. Apparently, the delta mu H+ dependency of in vitro protein translocation varies with the signal sequence core-region composition. Furthermore, a proline residue at position -10 resulted in a signal sequence that did not prevent the folding of the precursor in an in vitro trimerization assay. PMID- 8730865 TI - mRNA sequences influencing translation and the selection of AUG initiator codons in the yeast Saccharomyces cerevisiae. AB - The secondary structure and sequences influencing the expression and selection of the AUG initiator codon in the yeast Saccharomyces cerevisiae were investigated with two fused genes, which were composed of either the CYC7 or CYC1 leader regions, respectively, linked to the lacZ coding region. In addition, the strains contained the upf1-delta disruption, which stabilized mRNAs that had premature termination codons, resulting in wild-type levels. The following major conclusions were reached by measuring beta-galactosidase activities in yeast strains having integrated single copies of the fused genes with various alterations in the 89 and 38 nucleotide-long untranslated CYC7 and CYC1 leader regions, respectively. The leader region adjacent to the AUG initiator codon was dispensable, but the nucleotide preceding the AUG initiator at position -3 modified the efficiency of translation by less than twofold, exhibiting an order of preference A > G > C > U. Upstream out-of-frame AUG triplets diminished initiation at the normal site, from essentially complete inhibition to approximately 50% inhibition, depending on the position of the upstream AUG triplet and on the context (-3 position nucleotides) of the two AUG triplets. In this regard, complete inhibition occurred when the upstream and downstream AUG triplets were closer together, and when the upstream and downstream AUG triplets had, respectively, optimal and suboptimal contexts. Thus, leaky scanning occurs in yeast, similar to its occurrence in higher eukaryotes. In contrast, termination codons between two AUG triplets causes reinitiation at the downstream AUG in higher eukaryotes, but not generally in yeast. Our results and the results of others with GCN4 mRNA and its derivatives indicate that reinitiation is not a general phenomenon in yeast, and that special sequences are required. PMID- 8730864 TI - Identification of a second family of high-molecular-weight adhesion proteins expressed by non-typable Haemophilus influenzae. AB - We previously reported that two surface-exposed high-molecular-weight proteins, HMW1 and HMW2, expressed by a prototypic strain of non-typable Haemophilus influenzae (NTHI), mediate attachment to human epithelial cells. These proteins are members of a family of highly immunogenic proteins common to 70-75% of NTHI strains. NTHI strains that lack HMW1/HMW2-like proteins remain capable of efficient attachment to cultured human epithelial cells, suggesting the existence of additional adhesion molecules. We reasoned that characterization of high molecular-weight immunogenic proteins from an HMW1/HMW2-deficient strain might identify additional adhesion proteins. A genomic library was prepared in lambda EMBL3 with chromosomal DNA from non-typable Haemophilus strain 11, a strain that lacks HMW1/HMW2-like proteins. The library was screened immunologically with convalescent serum from a child naturally infected with strain 11, and phage clones expressing high-molecular-weight recombinant proteins were identified by Western blot analysis. One clone was identified that expressed a protein with an apparent molecular mass greater than 200 kDa. Transformation of non-adherent Escherichia coli strain DH5 alpha with plasmids containing the genetic locus encoding this protein gave rise to E. coli transformants that adhered avidly to Chang conjunctival cells. Subcloning and mutagenesis studies localized the DNA conferring the adherence phenotype to a 4.8 kbp fragment, and nucleotide sequence analysis further localized the gene encoding the adhesion protein to a 3.3 kbp open reading frame predicted to encode a protein of 114 kDa. The gene was designated hia for Haemophilus influenzae adhesin. Southern analysis revealed an hia homologue in 13 of 15 HMW1/HMW2-deficient non-typable H. influenzae strains. In contrast, the hia gene was not present in any of 23 non-typable H. influenzae strains which expressed HMW1/HMW2-like proteins. Identification of this second family of high-molecular-weight adhesion proteins suggests the possibility of developing vaccines based upon a combination of HMW1/HMW2-like proteins and Hia like proteins which would be protective against disease caused by most or all non typable H. influenzae. PMID- 8730866 TI - Generation of Campylobacter fetus S-layer protein diversity utilizes a single promoter on an invertible DNA segment. AB - Wild-type strains of Campylobacter fetus contain a monomolecular array of surface layer proteins (SLPs) and vary the antigenicity of the predominant SLP expressed. Reciprocal recombination events among the eight genomic SLP gene cassettes, which encode 97- to 149 kDa SLPs, permit this variation. To explore whether SLP expression utilizes a single promoter, we created mutant bacterial strains using insertional mutagenesis by rescue of a marker from plasmids. Experimental analysis of the mutants created clearly indicates that SLP expression solely utilizes the single sapA promoter, and that for variation C. fetus uses a mechanism of DNA rearrangement involving inversion of a 6.2 kb segment of DNA containing this promoter. This DNA inversion positions the sapA promoter immediately upstream of one of two oppositely oriented SLP gene cassettes, leading to its expression. Additionally, a second mechanism of DNA rearrangement occurs to replace at least one of the two SLP gene cassettes bracketing the invertible element. As previously reported promoter inversions in prokaryotes, yeasts and viruses involve alternate expression of at most two structural genes, the ability of C. fetus to use this phenomenon to express one of multiple cassettes is novel. PMID- 8730867 TI - Saccharomyces cerevisiae TEC1 is required for pseudohyphal growth. AB - Diverse eukaryotic organisms share developmental transcription factors with homologous DNA-binding domains. We showed that the developmental regulator AbaA, a member of the ATTS/TEA (AbaA, TEF-1, TEC1, Scalloped/TEF-1, TEC1, AbaA) class of transcription factors of the filamentous fungus Aspergillus nidulans, induces pseudohyphal development in the yeast Saccharomyces cerevisiae. The S. cerevisiae homologue of AbaA, TEC1p, is required for this morphological transition. We provide evidence that TEC1p functions in co-operation with STE12p to induce pseudohyphal development. PMID- 8730868 TI - Characterization of the distal promoter element of halobacteria in vivo using saturation mutagenesis and selection. AB - The sequence and spacing requirements of the archaeal "distal promoter element' (DPE) were examined by randomizing positions -19 to -32 upstream of the transcriptional start site of the ferredoxin (fdx) promoter of Halobacterium salinarium. This randomized promoter library containing 4(14) entries was cloned in front of the dihydrofolate reductase (DHFR) reporter gene and transformed into Haloferax volcanii. Two approaches were used to characterize these synthetic promoters. First, 1040 independent clones were randomly chosen and their degrees of trimethoprim resistance were determined. The sequences of 20 clones that were either sensitive, partially resistant or very resistant, respectively, were determined. Secondly, the transformed library was screened by direct selection for high-activity promoters by growing transformants in the presence of trimethoprim. Both approaches produced the following consensus sequence for a halobacterial promoter: (Formula: see text) (where R = A or G; Y = C or T; W = A or T; S = G or C; N = A, C, G or T). Further characterization of two sensitive, two partially resistant, and two very resistant clones verified that DHFR activity and cell phenotype are directly correlated. Sensitive clones did not contain detectable dhfr mRNA, whereas partially resistant clones contained a 700 nucleotide (nt)-long transcript, and very resistant clones contained both the 700nt-long transcript and a second, more abundant, 500nt-long truncated transcript. Quantification of the dhfr mRNA and DHFR enzyme activity suggests that the 3'-untranslated region of the dhfr transcript, missing from the shorter transcript, functions as a negative regulator of translation. PMID- 8730869 TI - Effects of F-encoded components and F-pilin domains on the synthesis and membrane insertion of TraA'-'PhoA fusion proteins. AB - F-pilin, the 70-amino-acid F-pilus subunit, accumulates in the cell envelope of F+ strains in a process that requires interactions between its precursor (the traA gene product) and other host and F-encoded proteins. Here, we have used a set of phi(traA-phoA) genes to explore the effects of different TraA domains on the synthesis and membrane insertion of TraA-PhoA fusion proteins, particularly in relation to other F-encoded gene products. The 51-amino-acid TraA leader peptide fused directly to alkaline phosphatase was synthesized at comparable rates and incorporated rapidly and efficiently into the inner membrane in F' and F- cells. A second fusion gene encoded the TraA leader peptide and the first 51 amino acids of F-pilin itself fused to PhoA (TraA'-'PhoA-102 polypeptide). Alkaline phosphatase activities and patterns of pulse-labelled polypeptides indicated that TraA'-'PhoA-102 was synthesized at comparable rates in F' and F- cells, but in neither was the TraA'-'PhoA-102 polypeptide efficiently processed as a membrane protein. A third gene encoded the entire 121-amino-acid TraA polypeptide fused to PhoA (TraA-'PhoA-121 polypeptide). About 70% of the pulse labelled TraA-'PhoA-121 polypeptide was rapidly processed in F' cells, where it accumulated in the cell envelope as active alkaline phosphatase, whereas in F- cells, < 5% of the pulse-labelled polypeptide was processed. Additionally, the apparent rate of TraA-'PhoA-121 polypeptide synthesis was threefold higher in F' cells. The traQ gene alone could not substitute for F in restoring TraA-'PhoA-121 (or wild-type F-pilin) accumulation. PMID- 8730870 TI - A periplasmic protein (Skp) of Escherichia coli selectively binds a class of outer membrane proteins. AB - A search was performed for a periplasmic molecular chaperone which may assist outer membrane proteins of Escherichia coli on their way from the cytoplasmic to the outer membrane. Proteins of the periplasmic space were fractionated on an affinity column with sepharose-bound outer membrane porin OmpF. A 17 kDa polypeptide was the predominant protein retained by this column. The corresponding gene was found in a gene bank; it encodes the periplasmic protein Skp. The protein was isolated and it could be demonstrated that it bound outer membrane proteins, following SDS-PAGE, with high selectivity. Among these were OmpA, OmpC, OmpF and the maltoporin LamB. The chromosomal skp gene was inactivated by a deletion causing removal of most of the signal peptide plus 107 residues of the 141-residue mature protein. The mutant was viable but possessed much-reduced concentrations of outer membrane proteins. This defect was fully restored by a plasmid-borne skp gene which may serve as a periplasmic chaperone. PMID- 8730871 TI - Identification and characterization of a novel type of replication terminator with bidirectional activity on the Bacillus subtilis theta plasmid pLS20. AB - We have sequenced and analysed a 3.1 kb fragment of the 55 kb endogenous Bacillus subtilis plasmid pLS20 containing its replication functions. Just outside the region required for autonomous replication, a segment of 18 bp was identified as being almost identical to part of the major B. subtilis chromosomal replication terminator. Here, we demonstrate that this segment is part of a functional replication terminator. This newly identified element, designated TerLS20, is the first replication terminator identified on a theta plasmid from a Gram-positive bacterium. TerLS20 is distinct from other known replication terminators in the sense that it is functional in both orientations. The region required for bipolar functionality of TerLS20 was delineated to a sequence of 29 bp, which is characterized by an imperfect dyad symmetry. PMID- 8730872 TI - Isolation of periplasmic nitrate reductase genes from Rhodobacter sphaeroides DSM 158: structural and functional differences among prokaryotic nitrate reductases. AB - The phototrophic bacterium Rhodobacter sphaeroides DSM 158 has a periplasmic nitrate reductase which is induced by nitrate and it is not repressed by ammonium or oxygen. In a Tn5 mutant lacking nitrate reductase activity, transposon insertion is localized in a 1.2 kb EcoRI fragment. A 0.6 kb BamHI-EcoRI segment of this region was used as a probe to isolate, from the wild-type strain, a 6.8 kb PstI fragment carrying the putative genes coding for the periplasmic nitrate reductase. In vivo protein expression and DNA sequence analysis reveal the presence in this region of three genes, napABC, probably organized in an operon. These genes are required for nitrate reduction, as deduced by mutational and complementation studies. The napA gene codes for a protein with a high homology to the periplasmic nitrate reductase from Alcaligenes eutrophus and, to a lesser extent, to other prokaryotic nitrate reductases and molybdenum-containing enzymes. The napB gene product has two haem c-binding sites and shows a high homology with the cytochrome c-type subunit of the periplasmic nitrate reductase from A. eutrophus. NAPA and NAPB proteins appear to be translated with signal peptides of 29 and 24 residues, respectively, indicating that mature proteins are located in the periplasm. The napC gene codes for a 25 kDa protein with a transmembrane sequence of 17 hydrophobic residues. NAPC has four haem c-binding sites and is homologous to the membrane-bound c-type cytochromes encoded by Pseudomonas stutzeri nirT and Escherichia coli torC genes. The phenotypes of defined insertion mutants constructed for each gene also indicate that periplasmic nitrate reductase from R. sphaeroides DSM 158 is a dimeric complex of a 90 kDa catalytic subunit (NAPA) and a 15 kDa cytochrome c (NAPB), which receives electrons from a membrane-anchored tetrahaem protein (NAPC), thus allowing electron flow between membrane and periplasm. This nitrate-reducing system differs from the assimilatory and respiratory bacterial nitrate reductases at the level of cellular localization, regulatory properties, biochemical characteristics and gene organization. PMID- 8730873 TI - The Escherichia coli ribosomal protein S16 is an endonuclease. AB - The histone-like protein HU isolated from Escherichia coli exhibited, after several purification steps, a Mg(2+)-dependent nuclease activity. We show here that this activity can be dissociated from HU by a denaturation-renaturation step, and is due to a small fraction of ribosomal protein S16 co-purifying with HU. S16 is an essential component of the 30S ribosomal particles. We have cloned, overproduced, and purified a histidine-tagged S16 and shown that this protein is a DNA-binding protein carrying a Mg(2+)-Mn(2+)-dependent endonuclease activity. This is an unexpected property for a ribosomal protein. PMID- 8730874 TI - Inducible gene expression mediated by a repressor-operator system isolated from Lactococcus lactis bacteriophage r1t. AB - A regulatory region of the temperate Lactococcus lactis bacteriophage r1t chromosome has been cloned and characterized. It encompasses the two divergently oriented genes rro, encoding the phage repressor, and tec. Both genes, of which the transcription start sites have been mapped, are preceded by consensus -35 and -10 promoter sequences. The region contains three 21 bp direct repeats with internal dyad symmetry which probably act as operators. Two of these repeats partially overlap the two promoter sequences. The distant third repeat is located within the tec coding sequence. Gel mobility shift assays demonstrated that Rro specifically binds to this sequence. To study possible transcriptional regulation of the region, a lacZ translational fusion with an open reading frame following tec was constructed. Under conditions that favour the lysogenic life cycle of r1t, beta-galactosidase activity was very low. Expression of the lacZ fusion could be induced 70-fold by the addition of mitomycin C at a concentration which promotes the switch of r1t from the lysogenic to the lytic life cycle. In non induced cells, promoter activity was repressed by Rro, as a frameshift mutation in rro resulted in constitutive expression of the lacZ gene fusion. PMID- 8730875 TI - Sequence analysis and molecular characterization of the temperate lactococcal bacteriophage r1t. AB - The temperate lactococcal bacteriophage r1t was isolated from its lysogenic host and its genome was subjected to nucleotide sequence analysis. The linear r1t genome is composed of 33,350 bp and was shown to possess 3' staggered cohesive ends. Fifty open reading frames (ORFs) were identified which are, probably, organized in a life-cycle-specific manner. Nucleotide sequence comparisons, N terminal amino acid sequencing and functional analyses enabled the assignment of possible functions to a number of DNA sequences and ORFs. In this way, ORFs specifying regulatory proteins, proteins involved in DNA replication, structural proteins, a holin, a lysin, an integrase, and a dUTPase were putatively identified. One ORF seems to be contained within a self-splicing group I intron. In addition, the bacteriophage att site required for site-specific integration into the host chromosome was determined. PMID- 8730876 TI - Tetrapac (tpc), a novel genotype of Neisseria gonorrhoeae affecting epithelial cell invasion, natural transformation competence and cell separation. AB - We characterized a novel mutant phenotype (tetrapac, tpc) of Neisseria gonorrhoeae (Ngo) associated with a distinctive rough-colony morphology and bacterial growth in clusters of four. This phenotype, suggesting a defect in cell division, was isolated from a mutant library of Ngo MS11 generated with the phoA minitransposon TnMax4. The tpc mutant shows a 30% reduction in the overall murein hydrolase activity using Escherichia coli murein as substrate. Tetrapacs can be resolved by co-cultivation with wild-type Ngo, indicating that Tpc is a diffusible protein. Interestingly, Tpc is absolutely required for the natural transformation competence of piliated Ngo. Mutants in tpc grow normally, but show a approximately 10-fold reduction in their ability to invade human epithelial cells. The tpc sequence reveals an open reading frame of approximately 1 kb encoding a protein (Tpc) of 37 kDa. The primary gene product exhibits an N terminal leader sequence typical of lipoproteins, but palmitoylation of Tpc could not be demonstrated. The ribosomal binding site of tpc is immediately downstream of the translational stop codon of the folC gene coding for an enzyme involved in folic acid biosynthesis and one-carbon metabolism. The tpc gene is probably co transcribed from the folC promoter and a promoter located within the folC gene. The latter promoter sequence shares significant homology with E. coli gearbox consensus promoters. All three mutant phenotypes, i.e. the cell separation defect, the transformation deficiency and the defect in cell invasion can be restored by complementation of the mutant with an intact tpc gene. To some extent the tcp phenotype is reminiscent of iap in Listeria, lytA in Streptococcus pneumoniae and lyt in Bacillus subtilis, all of which are considered to represent murein hydrolase defects. PMID- 8730877 TI - Mutational analysis of the Escherichia coli spoT gene identifies distinct but overlapping regions involved in ppGpp synthesis and degradation. AB - The spoT gene of Escherichia coli encodes a guanosine 3',5'-bis(diphosphate) 3' pyrophosphohydrolase (ppGppase) as well as an apparent guanosine 3',5' bis(diphosphate) synthetase (designated PSII). To determine the regions of the SpoT protein that are required for these two competing activities, we analysed plasmid-borne deletion mutations for their ability to complement chromosomal mutations defective in each activity. We found that a region containing the first 203 amino acids of the 702-amino-acid SpoT protein was sufficient for ppGppase activity while an overlapping region containing residues 67-374 was sufficient for PSII activity. These data indicate that the catalytic sites involved in the two activities are separate but closely linked in the primary sequence of the SpoT protein. A ppGppase-defective delta 1-58 deletion mutant strain failed to synthesize ppGpp in response to nutrient limitation, also supporting the notion that PSII activity from wild-type SpoT does not increase in response to nutrient limitation. Using a strain lacking PSII activity but retaining ppGppase activity, we determined the contribution of the RelA protein (ppGpp synthetase I, PSI) to ppGpp synthesis following glucose starvation. We found that the RelA protein activity accounts for the initial burst of ppGpp synthesis at the onset of glucose starvation but that this source of synthesis is absent when amino acids are present during glucose starvation. PMID- 8730879 TI - Hepatocyte growth factor/scatter factor and the placenta. PMID- 8730878 TI - Not trophoblast alone: a review of the contribution of the fetal microvasculature to transplacental exchange. AB - The fetal microcirculation of the term human placenta offers an interesting microvascular model. A perfused placenta can be used for integrated studies of vascular permeability-structure relationships. The organization of the paracellular pathway in human placental microvessels closely resembles not only that of the guinea-pig placenta, but also that seen in typical continuous non cerebral capillaries such as those of the myocardium. This uniformity of organization has allowed the development of a model of the organization of endothelial junctional complexes that allows testable predictions about the relationship between junctional organization and microvascular permeability. The key features of this model are: (1) molecular size restriction may be determined by a fibre matrix based on cadherin arrays in the zonula adhaerens. (2) The zonula occludens (tight junction) is discontinuous and so cannot act as a molecular sieve for solutes. It may serve as a shutter that limits the proportion of the paracellular cleft available for permeation. The main implication for placental function is that the human placental microcirculation is relatively tight and is an important restriction to diffusive permeation of the maternal fetal barrier by large molecules. PMID- 8730880 TI - Angiotensin II and its different receptor subtypes in placenta and fetal membranes. AB - The recent discovery of a local renin-angiotensin system in trophoblastic tissues has raised many questions regarding its role in the physiology of normal gestation and its implications in the pathophysiology of hypertension during pregnancy. In this article, the authors first review the most interesting aspects of the chorioplacental renin-angiotensin system, dwelling on the tissue distribution of angiotensin II and its receptor subtypes in the placenta and fetal membranes of different species. The relationship between angiotensin II and other locally synthesized chorioplacental substances is also analysed and the therapeutic implications of phenomena observed in pregnancy-associated hypertension are discussed. PMID- 8730881 TI - Immunohistochemical localization of vascular endothelial growth factor in the human placenta. AB - To understand the role of vascular endothelial growth factor (VEGF) in placental development, we examined immunohistochemically 56 placentae ranging from 6--41 weeks gestation using rabbit antibody to a synthetic multiple antigen peptide (MAP), composed of N-terminal amino acid residues 1--20 of human VEGF. In the present study, syncytiotrophoblast and invading extravillous trophoblasts ubiquitously expressed VEGF throughout gestation. However, the expression of VEGF in syncytiotrophoblasts was uneven in the first trimester and most intense at the sprouting sites. In addition, some stromal cells in the villi and decidual cells were also positive in the first trimester. A morphometrical analysis of the ratio of the VEGF-positive cell area to the capillary area in the terminal villi statistically revealed a critical point of change at 16 weeks' gestation. These results provide further evidence to support the hypothesis that VEGF, locally expressed by trophoblasts, stromal cells in villi and decidual cells, may play an important role in the physiological growth and function of the vascular system in the villous stroma and basal plate during placental development and maturation. PMID- 8730882 TI - Keratinocyte growth factor and its receptor in the rhesus macaque placenta during the course of gestation. AB - Keratinocyte growth factor (KGF) is synthesized and secreted exclusively by mesenchymal cells, and acts through its receptor (KGFR) to stimulate epithelial proliferation. In vivo, KGF and KGFR comprise a mesenchymal-epithelial cell paracrine system that can mediate epithelial cell mitosis. In preliminary work, we noted that KGF was expressed in the rhesus monkey placenta, and we now report on the expression of placental KGF and KGFR mRNAs during the course of gestation in this species. In-situ hybridization revealed that during early gestation, KGF mRNA was strongly expressed in placental mesenchymal cells. These cells, which were also immunoreactive for vimentin, were mainly located on the periphery of the mesenchymal cores of both anchoring and floating villi. KGFR mRNA was expressed in the adjacent trophoblastic epithelium, which was immunoreactive for cytokeratin. In-situ hybridization revealed that KGF mRNA expression was very high in the youngest placentae (34-days gestation) and decreased gradually to minimal levels by late gestation (157 days). Northern blot analysis indicated also that the KGF MRNA signal was strongest in early gestation samples and weakest by late gestation. Analysis for KGFR mRNA by a reverse transcriptase polymerase chain reaction technique showed that KGFR mRNA expression could be detected at all stages. However, in-situ hybridization indicated that KGFR mRNA expression was highest in early gestation placentae and least in the oldest placentae. Autoradiographs of frozen sections of placenta that had been incubated with [125I]KGF to detect receptor binding showed that grain density over the trophoblast was highest in the youngest and least in the oldest placentae. PCNA and Ki-67 expression followed this same temporal trend. We conclude that the KGF/KGFR system may be important in proliferation of the placental trophoblast during early- to mid-pregnancy in rhesus monkeys. PMID- 8730883 TI - Partial characterization of an immortalized human trophoblast cell-line, TCL-1, which possesses a CSF-1 autocrine loop. AB - Many previous studies in both mouse and human placenta have implicated a role for colony stimulating factor-1 (CSF-1) in the regulation of placental development. In this study we have examined CSF-1 production by an immortalized cell line (TCL 1) derived from the choriodecidua, transfected with a retrovirus gene coding for the large-T antigen. TCL-1 cells were uniformly positive by immunocytochemistry for the composite sub-units of human chorionic g gonadotrophin (hCG) but were negative for markers of other cell types localized at the fetal-maternal interface. Gelatinase enzymes were secreted by TCL-1 cells cultured on extracellular matrix in a manner indicative of extra-villous trophoblast. Dot blot immunoassays and ELISA indicated that CSF-1 was secreted by TCL-1 cells, at levels comparable to primary trophoblast cells and BeWo choriocarcinoma (trophoblast tumour) cells. Reverse transcriptase-polymerase chain reaction analysis confirmed the presence in TCL-1 cells of CSF-1 receptor mRNA (c-fms gene product), indicating that the components of a potential autocrine loop were present in these cells. Proliferation of TCL-1 cells was not affected by the addition of exogenous CSF-1 but was elevated in response to treatment with a CSF 1 neutralizing antibody. The immortalized cell line, TCL-1, provides a potential model in which to investigate regulation of growth and differentiation of trophoblast cells in vitro. PMID- 8730884 TI - High densities of angiotensin II subtype 2 (AT2) receptors in the porcine placenta and fetal membranes. AB - The present study was undertaken in order to characterize and determine angiotensin (Ang) II receptors and renin in the porcine placenta and fetal membranes. High densities of Ang II receptors of subtype 2 (AT2 receptors) were demonstrated in all parts of the placenta and fetal membranes throughout gestation. AT1 receptors (subtype 1) were only found in the first part of the gestation and only in the allantochorion-endometrium at low densities. The total Ang II receptor density was increased in the allantochorion-endometrium in the last stage of gestation. Except for this finding the Ang II receptor density did not vary during gestation. Some of the highest receptor densities were found in the allantoamnionic membrane. The Ang II receptor densities did not correlate with the duration of the gestation in any part of the placenta. Enzymatically active renin was found in all parts of the porcine placenta and fetal membranes in concentrations similar to those found earlier in the blood plasma. The renin concentrations did not correlate with the Ang II receptor densities or the duration of the gestation in any part of the placenta. Compared with the human placenta, where the Ang II receptors are almost exclusively AT1 receptors and low receptor densities are found in the fetal membranes, the present results indicate profound species differences in the expression and function of the renin angiotensin system in the placenta and fetal membranes. PMID- 8730885 TI - Histochemical evaluation of placental angiotensinase A in pre-eclampsia: enzyme activity in villous trophoblast indicates an enhanced likelihood of gestational proteinuric hypertension. AB - The aim of this study was to examine whether differences in placental angiotensinase A (glutamyl aminopeptidase, EC 3.4.11.7) activities occurred in hypertensive complications of pregnancy compared with uncomplicated pregnancies. Biochemical and semiquantitative histochemical methods were used and compared for their applicability. Angiotensinase A activity was detected using L-alpha glutamyl-4-methoxy-2-naphthylamide (alpha-Glu-MNA) as substrate and Fast Blue B salt for simultaneous azo-coupling in cryostat sections of placental tissue samples from 32 patients with pre-eclampsia, 11 patients with pregnancy-induced hypertension and 44 participants with uncomplicated pregnancies. The graduated intensity of reaction product in the villous trophoblast and in fetal blood vessels was evaluated semiquantitatively in a double-blind study by light microscopy (semiquantitative score method). Score levels were related to relative frequencies of hypertensive disorders (proportional odds model) and correlated to the severity of gestational hypertension (Spearman's rank correlation). After detection of enzyme activity, the same tissue samples were homogenized and used for kinetic fluorometric measurements under the same substrate and buffer conditions as in enzyme histochemistry. Enhanced villous trophoblastic angiotensinase A activity was significantly associated with an increased frequency of pre-eclampsia in pregnant women (cumulative odds ratio x 0(1) 6.37; P < 0.001) and showed significant correlations with the severity of gestational hypertensive disorders, represented by systolic (r = 0.31; P < 0.05) and diastolic (r = 0.34; P < 0.05 blood pressure and by concomitant proteinuria (r = 044; P < 0.01). Histochemical evaluation of fetal blood vessels and biochemical measurements revealed no statistically significant results. In conclusion this study demonstrates for the first time that increased villous trophoblastic angiotensinase A activity indicates an increased likelihood of the presence of pre-eclampsia and the severity of hypertensive disorders in pregnancy. PMID- 8730886 TI - Nitric oxide concentrations are increased in the feto-placental circulation in intrauterine growth restriction. AB - The aim of this study was to measure plasma concentrations of total nitrites, as an index of nitric oxide (NO) synthesis, in the fetal circulation of normal pregnancies and in pregnancies complicated by intrauterine growth restriction. Plasma was prepared from umbilical venous blood collected from 13 placentae from normal pregnancies complicated by intrauterine growth restriction. Plasma NO concentrations were determined using the Greiss reaction by measuring combined oxidation products of NO, plasma nitrite (NO2-) and nitrate (NO3-) after reduction with nitrate reductase. Significantly higher NO2-concentrations were found in umbilical venous plasma in the group complicated by intrauterine growth restriction compared to the control group (65.6 mumol/1, P < 0.001. These results support the hypothesis that increased NO production may be a compensatory response to improve blood flow in the placenta and/or may play a role in limiting platelet adhesion and aggregation. PMID- 8730887 TI - Protein intake in pregnancy, placental glucocorticoid metabolism and the programming of hypertension in the rat. AB - Hypertension is strongly predicted by a low birthweight:placental weight ratio. Two independent models have been described to explain this association; less than optimal maternal protein nutrition leading to fetal undernutrition, or glucocorticoid excess. Pregnant rats were fed diets containing 18 per cent casein (control) or 9 per cent casein, balanced for energy. On day 20 of gestation the pregnancies were terminated and placentae collected for determination of 11 beta hydroxysteroid dehydrogenase (11 beta HSD) activity. Placental 11 beta HSD normally protects the fetus from the effects of maternal glucocorticoids. Activity was specifically attenuated by mild protein restriction (33 per cent in activity), whilst activities of glucocorticoid-insensitive control enzymes were unchanged and glucocorticoid-inducible glutamine synthetase activity was increased (27 per cent), relative to activity in placentae from control animals. The nutritional manipulation during pregnancy significantly increased systolic blood pressure (17 mmHg) in the resulting offspring in early adulthood. A possible common pathway whereby maternal environmental factors may influence fetal and placental growth and programme disease is inferred. PMID- 8730888 TI - Dipeptide transport in brush-border membrane vesicles (BBMV) prepared from human full-term placentae. AB - The uptakes of the tritiated, hydrolysis-resistant cationic (d-Phe-L-Lys), neutral (D-Phe-L-Ala) and anionic (D-Phe-L-Glu) peptides into human full-term placental brush-border membrane vesicles (BBMV) were time-dependent and into an osmotically-active space. Uptakes of D-Phe-L-Lys and D-Phe-L-Glu were temperature dependent. Uptake of D-Phe-L-Lys was electroneutral (either cation exchange or anion co-transport), whereas D-Phe-L-Ala and D-Phe-L-Glu were both stimulated by an increasingly inside-positive membrane potential (explained by either cation exchange or anion co-transport, or translocation alone, respectively). Uptake of D-Phe-L-Ala was stimulated (approximately 50 per cent) by an inwardly-directed proton gradient (pHin = 7.4, pHout = 5.5), whereas D-Phe-L-Glu was unaffected, and D-Phe-L-Lys uptake was inhibited (approximately 50 per cent) but was unaffected by the organic cation-exchange inhibitors 1,1-diethyl-2,2-cyanine (decynium22) and 5-(N-methyl-N-isobutyl)amiloride (MIBA). Over the concentration range studies, the peptides did not self-inhibit, and the only cross-inhibition was by D-Phe-L-Glu on D-Phe-L-Lys uptake (estimated K(I) 24.2 +/- 1.36 mM), suggesting very low affinity transporter(s). Under conditions favouring its transport by PepT1, D-Phe-L-Glu uptake was unaffected by diethylpyrocarbonate (DEPC); neither D-Phe-L-Ala nor D-Phe-L-Lys was inhibited by DEPC under maximally proton-stimulated conditions of uptake. We conclude that Pep-T-like transporters are not responsible for peptide uptake into human placental BBMV; while the molecular identity of the transporter(s) involved remains unclear, we hypothesize that they could be similar to the as yet unidentified epithelial basolateral peptide transporter(s). PMID- 8730889 TI - Distribution of annexin I and II in term human fetal membranes, decidua and placenta. AB - Annexin I and II are calcium binding proteins implicated in the regulation of a number of cellular functions, including secretory processes, prolactin release and prostaglandin formation. The cellular distribution of these proteins was examined in human term placenta, fetal membranes (amnion and chorion laeve) and decidua using immunohistochemistry. Annexin I was found in amnion epithelial cells and chorion laeve trophoblast but not in decidua, and was located in the syncytiotrophoblast cells of placenta. Annexin II and annexin II light-chain were located in the amnion epithelial cells, the cells of the mesenchymal layer between the amnion and chorion laeve trophoblast and endothelial cells lining the blood vessels in the decidua. In contrast to annexin I, annexin II was located in the villous core and not the syncytiotrophoblast cells in the placenta. There was no apparent change in the distribution of these annexins during labour. These findings indicate that the cellular distribution of these annexins is different, and may be an important consideration when examining their synthesis or action in tissues and in vitro with mixed cell populations and tissue homogenates. PMID- 8730891 TI - Volume-activated release of D-aspartate from human placental tissue. PMID- 8730890 TI - The increased endotoxin-sensitivity of pregnant rats, as reflected by glomerular ecto-ADP-ase activity, is not dependent on the presence of decidual cells. AB - In the present study the possible role of decidual cells in the pregnancy associated increased sensitivity of glomerular ecto-ADP-ase to endotoxin was investigated. Early (day 5) pregnant (E-Pr; n = 10), pseudopregnant (E-PSP; n = 10), (day 5), pseudopregnant rats with a decidualized uterus (E-DEC; n - 10), as well as late (day 14) pregnant (L-Pr; n = 10), pseudopregnant (L-PSP: n = 10) (day 14), and pseudopregnant rats with a decidualized uterus (E-DEC; n = 10) were infused with either endotoxin (1.0 mg/kg bw) or saline. Three days later rats were killed and specimens of the left kidney were snap-frozen. Cryostat kidney sections (4 microns) were stained for ecto-ADP-ase activity and quantitatively evaluated. The results show that only glomerular ecto-ADP-ase activity of both groups of pregnant rats (E-Pr and L-Pr) was significantly decreased after endotoxin infusion as compared to saline infusion. In the other groups of rats, no significant differences in ecto-ADP-ase activity were observed between saline and endotoxin infusion. It is concluded that decidual cells do not play a role in the increased sensitivity of ecto-ADP-ase to endotoxin during pregnancy. PMID- 8730892 TI - Intracytoplasmic sperm injection in dysmorphic human oocytes. AB - Fertilisation and development of dysmorphic human oocytes recovered from hyperstimulated ovaries have been evaluated following intracytoplasmic sperm injection (ICSI) for treatment of male infertility. A total of 2968 oocytes at metaphase II of meiosis were injected, of which 806 (27.2%) were dysmorphic at the light microscopic level. Cytoplasmic abnormalities included granularity, areas of necrosis, organelle clustering, vacuoles, and accumulating saccules of smooth endoplasmic reticulum. Anomalies of the first polar body and zona pellucida, as well as non-spherical shapes of oocytes, were also noted. Contrary to previous findings linking some dysmorphisms to non-assisted fertilisation failure, in this study no single abnormality led to a reduction in the fertilisation rate, nor was fertilisation compromised in oocytes with multiple abnormalities. The incidence of normal fertilisation (two pronuclei and two polar bodies) was 69% in both the dysmorphic and non-dysmorphic oocytes. While overall pregnancy and implantation results were not altered in the group of patients (n = 242) in whom at least one dysmorphic oocyte was injected, exclusive replacement of embryos which originated from dysmorphic oocytes led to a higher incidence of early pregnancy loss. It is concluded that aberrations in the morphology of human oocytes--most probably a product of controlled ovarian stimulation--are of little or no consequence to fertilisation or early cleavage after ICSI. It is possible, however, that these embryos have a reduced potential for implantation and further development. PMID- 8730893 TI - Response of avian nuclei to mammalian maturation promoting factor (MPF). AB - Chicken blastodermal cells (stage X) were fused to mouse enucleated oocytes with either no or high maturation promoting factor (MPF) activity. High MPF levels always induced premature chromosome condensation (PCC) irrespective of the number of nuclei fused to a single oocyte. When a single blastodermal cell was fused to a single oocyte without MPF activity the nucleus remained intact for up to 3 h and thereafter underwent PCC. A quite different situation was observed after multiple fusion of several blastodermal cells to a single oocyte without MPF activity. Here, the transplanted nuclei remained intact even after prolonged culture but underwent extensive swelling. DNA synthesis was detected in almost all unfused blastodermal cells. However, after the fusion of several blastodermal cells to a single oocyte no DNA synthesis could be detected. These results provide further evidence that MPF is the universal cell-cycle regulator in the animal kingdom. Its activity is blocked (or neutralised) after fusion to several S-phase cells. Interestingly, our results further suggest that DNA synthesis is suppressed in meiotic cytoplasm even in the presence of an intact nuclear envelope. PMID- 8730894 TI - Comparison of the acrosome reaction-inducing ability of the outer and inner surfaces of the zona pellucida and oolemma: a study using the golden hamster. AB - The outer surface of hamster zona pellucida has a strong ability to induce the sperm acrosome reaction, whereas neither the inner surface of the zona nor the oolemma has this ability. Therefore, the spermatozoa that fertilise oocytes after subzonal insemination of spermatozoa (SUZI) must have been either acrosome reacted prior to injection or acrosome-reacted spontaneously while swimming within the perivitelline space. The outer surface of fresh mouse zona is not capable of inducing the acrosome reaction of hamster spermatozoa. When fixed with glutaraldehyde, however, it became capable of inducing the hamster sperm acrosome reaction rather efficiently. The reason for this is not clear. PMID- 8730896 TI - The microenvironment created by non-blocking embryos in aggregates may rescue blocking embryos via cell-embryo adherent contacts. AB - Under our culture conditions, mouse embryos from the BALB/c inbred mouse strain develop successfully in culture only from the late 2-cell stage onwards (so called 2-cell block), whether or not EDTA is added to the culture medium. (CBA x C57BL) F2 embryos do not exhibit a 2-cell block. Medium conditioned by culture of non-blocking embryos from the 2-cell to the 8-cell stage did not improve the development of blocking embryos, nor did co-culture of blocking and non-blocking embryos, with or without conditioned medium. On the other hand phytohaemagglutinin (PHA)-assisted aggregation of an early 2-cell BALB/c embryo with five surrounding non-blocking F2 embryos (2-cell or 8-cell) or five BALB/c 8 cell embryos allowed the early 2-cell BALB/c embryos to develop into blastocysts within 72 h. Aggregation of blocking BALB/c 2-cell embryos with each other had no 'rescue' effect. When blocking and non-blocking 2-cell embryos were aggregated together, an integrated blastocyst was formed; but when the early 2-cell BALB/c embryos were aggregated with non-blocking 8-cell embryos, the blocking embryos formed a separate small blastocyst, which nonetheless retained adherent contact with the non-blocking embryos throughout the culture period. Ultrastructural analysis showed that 2-cell embryos aggregated with the aid of PHA form close adherent cell contacts up to several micrometres in length. PMID- 8730895 TI - Absence of an intracellular pH change following fertilisation of the mouse egg. AB - The intracellular pH of the mouse egg was measured during fertilisation to determine whether an increase in pH accompanies activation of this mammalian egg. The pH-sensitive dye BCECF [2',7'-bis-(2-carboxyethyl)-5(and 6)carboxyfluorescein] was introduced into the mouse egg by incubation in BCECF-AM or by microinjection of dextran-conjugated BCECF. The cells were also loaded with the DNA-specific fluorochrome Hoechst 33342 to confirm fertilisation by observation of Hoechst-stained, decondensing sperm heads in the cytoplasm. The ratio of emission intensities for the dye (494/440 nm excitation wavelengths) was monitored continuously with a photon-counting photomultiplier tube. There was no change in pH during or after fertilisation. Control eggs displayed the expected increase in pH when exposed to NH4Cl. In other experiments, intracellular pH and intracellular Ca2+ were monitored simultaneously during fertilisation. The eggs were injected with BCECF dextran and Fura dextran. Fluorescence emission was recorded at excitation wavelengths of 495 nm (BCECF, pH-sensitive wavelength) and 385 nm (Fura, Ca(2+)-sensitive wavelength). A decrease in emission intensity at 385 nm excitation clearly marked the repetitive Ca2+ transients at egg activation. There was no change in the fluorescence emitted at 495 nm excitation, indicating an absence of any change in intracellular pH. These results indicate that intracellular alkalinisation of the cytoplasm does not accompany activation of this vertebrate egg. PMID- 8730897 TI - Association between p34cdc2 levels and meiotic arrest in pig oocytes during early growth. AB - The molecules involved in determining meiotic competence were determined in porcine oocytes isolated from preantral and antral follicles of different sizes. Oocytes isolated from preantral follicles had a mean diameter of 78 microns, contained diffuse filamentous chromatin in the germinal vesicle and were incapable of progressing from the G2 to the M phase of the cycle even after 72 h in culture. Oocytes from early antral follicles had a mean diameter of 105 microns, showed a filamentous chromatin configuration and about half resumed meiosis but arrested at metaphase I (MI) when cultured. Oocytes from mid-antral (3-4 mm) and large antral follicles (5-6 mm) had mean oocyte diameters of 115 and 119 microns respectively, contained condensed chromatin around the nucleolus and progressed to metaphase II (MII) in 48% and 93% of instances respectively. Analysis of p34cdc2, the catalytic subunit of maturation promoting factor (MPF), by immunoblotting indicates that the inability of small (78 microns) oocytes to resume meiosis is due, at least in part, to inadequate levels of the catalytic subunit of MPF. On the other hand, the inability of intermediate-sized (105 microns) oocytes from antral follicles to complete the first meiotic division by progressing beyond MI appears not to be limited by levels of p34cdc2, which are maximal by this stage. We postulate that an inadequacy of molecules other than p34cdc2 limits progression of MI to MII; the acquisition of these molecules during the final stages of growth may be correlated with the formation of the perinucleolar chromatin rim in the germinal vesicle. PMID- 8730898 TI - Hypotaurine and taurine in gamete and embryo environments: de novo synthesis via the cysteine sulfinic acid pathway in oviduct cells. AB - Taurine and hypotaurine seem to be important compounds for sperm survival and capacitation, the fertilisation process and embryo development, and are present in both sperm and genital secretions. Hypotaurine has protective effects against peroxidative damage. We have established a simple method for hypotaurine quantification in sperm and genital secretions. The mean concentration of hypotaurine is significantly higher in bovine than in human spermatozoa and in seminal plasma. We observed that both molecules are secreted by cow, sow, goat and rabbit oviduct epithelial cell monolayers. In rabbit the release is ascorbic acid dependent. Goat oviduct epithelial cells are able to use the transsulfuration pathway to form hypotaurine and taurine from methionine. We were able to identify cysteine sulfinate decarboxylase (EC 4.1.1.29) activity in cow and goat tubal monolayers. Our results demonstrate that hypotaurine and taurine are secreted by oviduct epithelium, and synthesised by tubal cells via the cysteine sulfinic acid pathway. The data obtained emphasise the importance of hypotaurine and taurine for gamete maturation, fertilisation and early embryonic development. PMID- 8730899 TI - Synergetic effects of epidermal growth factor and gonadotropins on the cytoplasmic maturation of pig oocytes in a serum-free medium. AB - Cumulus-enclosed or cumulus-free pig oocytes were cultured for 48 h in a serum free maturation medium, TCM-199B containing 1 mg/ml polyvinylalcohol, supplemented with gonadotropins (PMSG + hCG) and/ or epidermal growth factor (EGF), and inseminated with frozen-thawed ejaculated spermatozoa. Fourteen hours after insemination, the ability of oocytes to form male pronucleus (MPN) was examined. EGF significantly increased the ability of cumulus-enclosed oocytes to form MPN irrespective of the concentrations (1-100 ng/ml) tested. Gonadotropins had a greater effect than EGF on MPN formation in cumulus-enclosed oocytes, and a synergetic effect was observed when EGF and gonadotropins were added together. However, neither EGF nor gonadotropins exhibited a stimulatory effect on MPN formation in cumulus-free oocytes. These results indicate that the role(s) of both EGF and gonadotropins in stimulating cytoplasmic maturation of pig oocytes is mediated by cumulus. PMID- 8730900 TI - Estimation by radiation inactivation of the minimum functional size of acrosome reaction-inducing substance (ARIS) in the starfish, Asterias amurensis. AB - In the starfish Asterias amurensis, the jelly coat of the eggs contains a glycoprotein essential for the induction of the acrosome reaction in homologous spermatozoa that is termed the acrosome-reaction-inducing substance (ARIS). ARIS is a highly sulphated and fucose-rich glycoprotein of extremely high molecular mass (> 10(4) kDa). ARIS was irradiated with high-energy electrons in order to estimate the minimum size required for its biological activity. The minimum functional unit or target size of ARIS was estimated to be c. 14 kDa by target size analysis. ARIS was significantly disintegrated by the irradiation, yet the total sugar content was not apparently reduced. The binding of 125I-labelled ARIS to spermatozoa competed with that of irradiated ARIS, although the affinity of ARIS was much reduced after irradiation. PMID- 8730901 TI - The effect of temperature fluctuations on the cytoskeletal organisation and chromosomal constitution of the human oocyte. AB - The effect of temperature fluctuation on spindle integrity and chromosomal organisation in the human oocyte, and the consequences of such effects on the chromosomal constitution of resulting parthenotes, were investigated. A total of 340 oocytes were stained immunocytochemically with an antibody to alpha-tubulin, and 502 were activated parthenogenetically. Exposure of oocytes to room temperature for 2, 10 or 30 min caused disruption of the spindle in 77% (n = 26), 72% (n = 18) and 89% (n = 19) of cases respectively, with evidence of chromosomal dispersal in 50%, 56% and 52.6% respectively. These effects were reversed when oocytes were returned to 37 degrees C after exposure to room temperature for 2 min, but not after 10 min or 30 min. Temperature reduction affected rates of parthenogenetic activation of oocytes (2 min: 67%, n = 27; 10 min: 68%, n = 28; 30 min: 54%, n = 35) and cleavage of resulting parthenotes, but only if oocytes were exposed to room temperature for 30 min (30 min: 53%, n = 19). There is a direct association between temperature-induced spindle damage in the oocyte (70%, 50 of 63) and chromosomal abnormalities in parthenotes developed from oocytes exposed to room temperature (56%, 23 of 41; p < 0.01). PMID- 8730902 TI - The dynamics of professional commitment: immigrant physicians from the former Soviet Union in Israel. AB - The paper examines professional commitment among physicians who immigrated to Israel form the former Soviet Union during the early 1990s. This population faces severe limits regarding occupational continuity because of the highly saturated market in which non-negligible groups will, in the long run, of necessity undergo occupational change. The theoretical background for the analysis is drawn from the literature regarding recent changes in professional roles with particular reference to the shifting meaning of work in post-modern societies and its consequences for occupational commitment. The professional context of medical practice in the former Soviet Union and the social and economic constraints of Israeli society in the 1990s set the scene for the analysis. Several dimensions of professional commitment are examined empirically, on the assumption that there are a variety of ways to consider the notion of commitment and that no one measure tells a complete story. Prolonged processes of deprofessionalization of medicine in the Soviet Union, suggest that medicine for most immigrant physicians is not so much a 'calling' to which they are devoted; rather it is a necessary means to gain a livelihood, the only occupation for which they have been trained for many years after stringent selection to medical school and the only job in which they have worked consistently since completing their formal training. Two and a half years after arrival in Israel the immigrant doctors are characterized by a short-range time perspective which makes them unwilling to accept the constraints of the saturated market; intense efforts are made by most to obtain a license despite the fact that only a fraction of them will be able to work in their profession on a regular basis. Despite this over-riding reality, many hope that they will be among the selected few who will be able to obtain a medical post. PMID- 8730903 TI - Practice of in-vitro fertilization: a case study from Finland. AB - In 1978 the first human IVF-baby was born. Today IVF is a standard procedure in the treatment of infertility in industrialized Western countries. In this study we analyzed how IVF reached an established position as a medical innovation in Finland, how IVF-care was organized between 1991-1993, and which kind of women used IVF-services and delivered a child as a result. The data sources were interviews with practicing IVF-physicians, a survey of Finnish IVF-clinics, telephone interviews with a sample of the adult population, and data of mothers from the Finnish Birth Registry. IVF in Finland followed the four stages of a medical innovation from a promising report into a standard procedure. Key factors in the introduction of IVF-methods were the work of andrologists', and later, IVF physicians' associations, the approval of the method by head gynecologists in university clinics and among other colleagues, and later, the increase in IVF services without regulatory government policy. IVF has become increasingly available in private clinics because pioneer physicians have established such services. In principle there was no social discrimination in having IVF, because it was available almost free of charge in public clinics. But the costs and availability of private clinics created unequal access to IVF services. IVF-women were more often upper-class white collar employees living in southern Finland than women in the control group. IVF has been a routine treatment option of infertility since the end of the 1980s. It has provided a medical technology solution to infertility. The supply and demand of IVF has increased and its indications have widened in the treatment of infertility. This is the inner logic of a successful technology: after the developmental processes of a revolutionary innovation, the use of technology escalates rapidly and the barriers for its use decrease. PMID- 8730904 TI - The creation of protection and hope in patients with malignant brain tumours. AB - The malignant brain tumour disease condenses much of the anguish of cancer diseases. The brain is a vital and delicate organ, and the prognosis is generally unfavourable. The patient is exposed and has to rely on cognitive manoeuvres to manage the mental stress. The purpose of this study was to generate new insights into how the patient constructs a new sense of reality when confronted with the malignant brain tumour diagnosis. Within grounded theory methodology, 30 patients with malignant gliomas were interviewed twice, in direct connection with diagnosis, surgery and radiotherapy. In addition, their partners were interviewed, and quantitative instruments (SMMSE, RDCQ) were used as additional references for assessing the patients cognitively and emotionally. Eleven patients were excluded from the final analysis because of cognitive impairment of personality change. Most of the patients were aware of the fact that the brain tumour exposed them to grave danger, but they were also able to use various cognitive manoeuvres to create protection and hope. This process originated from different sources: the body; helpful relations; cognitive schemata; and the handling of information. The importance of the body to raise hope is emphasized. In the discussion we consider this process as an expression of how the patient brings together reality and hope, thus creating her/his own illusion. These findings are also related to adjacent psychoanalytic theory, proposing a theoretical reference with clinical implications when discussing "What to tell cancer patients." PMID- 8730905 TI - Forecasting the AIDS epidemic in Puerto Rico. AB - The purpose of this study is to model and predict the diffusion of the AIDS epidemic in Puerto Rico. Specifically we aim at identifying primary influences in the geographical distribution of the population affected with AIDS to produce a reasonable projection of the time and space paths that will be followed by the epidemic. The study is based on AIDS incidence data from 1982 through 1992. The epidemic is analyzed statistically through multivariate regression and the potential model to produce a probability surface in which risk behavior and intra municipality mobility are significant risk factors. The growth forecast is produced using the logistic function fitted to the past growth using an iterative non-linear optimization method. A simulation technique is then employed to forecast the spatial development of the epidemic from 1993 through 1997. The results indicate a clear hierarchical tendency at the beginning of the epidemic, later a wave-like diffusion pattern is also observed. While the absolute number of new cases is expected to remain higher in the more urbanized areas, the relative growth of AIDS cases is likely to become much higher in the rural municipalities. The forecasting procedure employed here is applicable to populations with diverse epidemiological profiles. PMID- 8730906 TI - Regulatory actions to enhance appropriate drug use: the case of antidiarrhoeal drugs. AB - Inappropriate drug use is a serious problem in the control of diarrhoeal diseases. To address this problem, the World Health Organization's Programme for the Control of Diarrhoeal Diseases reviewed the literature on the most commonly used antidiarrhoeal agents, and distributed the resulting document widely in 1990. Antidiarrhoeal drugs received simultaneous attention in the popular media as groups and individuals campaigned against their registration and use. This article evaluates the actions against antidiarrhoeal drugs taken by national drug regulators in the period during and after these events (January 1989 until December 1993). Information on regulatory actions was requested from countries and extracted from published and unpublished sources. Sixteen countries reported regulatory actions on 21 occasions during the period of study. The majority of actions concerned antimotility drugs; few were against adsorbents, antidiarrhoeal drugs containing antimicrobials, or adult formulae. Six countries took action against large and heterogenous groups of antidiarrhoeal drugs. Most actions occurred in the two-year period immediately following the distribution of the WHO review and the attention in the media. The sequence of distribution of the review, media coverage, and activities of dedicated groups and individuals, followed by a noticeable cluster of regulatory actions suggests a clear relationship. Further research is necessary to determine the relative role of each activity. There are several constraints to deregistration of profitable drugs and some drug regulators may have chosen to delay action until the end of the drug's registration cycle. Many more antidiarrhoeal drugs may lose their register in the future through a passive deregistration process. Deregistration of inappropriate drugs will probably take much time and widespread deregistrations are not likely. Furthermore, regulatory actions alone are probably insufficient to achieve a more appropriate use of drugs. More effect can be expected from simultaneous regulatory, managerial, and educational interventions directed at providers, combined with communication to the general public. PMID- 8730907 TI - Rationing health care: views from general practice. AB - General practitioners (GPs) in the United Kingdom are central to the commissioning of health care services. A qualitative study of their views was therefore designed, which incorporated an in-depth (open) interview technique carried out on a 20% sample of all GPs (n = 100) in one United Kingdom Health District. The data from these interviews indicated that GPs were aware of, but had mixed feelings about the need for rationing. They expressed disquiet about the dilemma faced in rationing health care at the time of the consultation and readily associated issues of cost in their practice with rationing. Some of the currently adopted methods of rationing (waiting lists, co-payments and ability to pay) were commented upon. The respondents also made suggestions on how rationing could be carried out, which included: maximizing efficiency to reduce the need for rationing; using a third party committee to make rationing decisions, with a membership of clinicians, managers, and possibly public representatives, and; being explicit about how rationing is done. Fundholding brought rationing decisions to the fore, and worried most who discussed it in the context of rationing. The conclusion of this paper is that current implicit rationing policies in the National Health Service are flawed as they assume that GPs will ration health care at the time of the consultation. The involvement of GPs in the rationing process is important (particularly given the present expansion of GP fundholding), so there is a need for an alternative to the present system. PMID- 8730909 TI - Exposure to missile attacks: the impact of the Persian Gulf War on physical health behaviours and psychological distress in high and low risk areas in Israel. AB - The purpose of this study was to compare residents of an area (Tel-Aviv), which was severely afflicted by SCUD missiles during the Persian Gulf War (high risk region), to residents of a low-risk region (Jerusalem) in terms of: (a) changes in physical health, in use of medical or psychological services, and in health behaviours during the period of the war compared to the preceding month; (b) levels of psychological distress (somatization and anxiety) during the war; (c) characteristics of persons at highest risk for psychological distress. Respondents were randomly chosen and interviewed by telephone (N = 545 in Tel Aviv, N = 406 in Jerusalem). The respondents in both regions reported significant yet similar deterioration in physical health status, and an increase in detrimental health behaviours during the Gulf War. Tel-Aviv residents had significantly higher levels of psychological distress as compared to residents of Jerusalem: in somatization 18 vs 12% respectively (OR = 2.44, CI = 1.39-4.28), in anxiety 34 vs 26% respectively (OR = 1.62, CI = 1.1-2.42). In addition to place of residence, age, ethnicity, religiosity and self-assessed health were identified as characteristics of persons at greater risk for psychological distress. PMID- 8730908 TI - An economic analysis of Chinese fertility behavior. AB - This paper is the first to present a Chinese general fertility model that simultaneously controls for the endogeneity of infant mortality and per capita income determination at county level. Using the 1982 Chinese population census data, comprising 2305 observational units, this analysis improves on existing studies in several ways. First, since all the underlying variables are measured at the Chinese county level, we treat both the per capita income and infant mortality rates as endogenous, as opposed to exogenous as assumed in most previous studies on Chinese fertility. Our testing results strongly reject the null hypothesis of the exogeneity of both infant mortality and income determination within our model. Secondly, concerning the hypothesis of nonlinear income effect on fertility behavior, we examine both the variable income elasticity and constant income-elasticity models. Strong evidence is obtained in support of the variable income elasticity model, predicting a U-shaped income effect on Chinese general fertility. This suggests that a more equitable income distribution leads to a reduction in the Chinese fertility rates. Thirdly, employing the two stage least squares procedure, we find a much stronger positive replacement effect on infant mortality when the endogeneity of infant mortality and income are both controlled for simultaneously. Our results indicate that Chinese general fertility may well be shaped by optimizing behavior. PMID- 8730910 TI - Distributing scarce livers: the moral reasoning of the general public. AB - The transplant system has been criticized for not paying enough attention to efficiency in distributing scarce organs. But little research has been done to see how the general public views tradeoffs between efficiency and equity. We surveyed members of the general public to see how they would distribute organs among patients with varying chances of benefiting from them. In addition, we asked subjects to explain their decisions and to tell us about any other information they would have liked in order to make the decisions. We found that the public places a very high value on giving everyone a chance at receiving scarce resources, even if that means a significant decrease in the chance that available organs will save people's lives. Our results raise important questions about whether the aims of outcomes research and cost-effective studies agree with the values of the general public. PMID- 8730911 TI - Local terminology for febrile illnesses in Bagamoyo District, Tanzania and its impact on the design of a community-based malaria control programme. AB - This paper reviews results of several ethnographic studies that have examined the issue of local terminology for malaria in Africa, then presents findings from an on-going study in Bagamoyo District, Tanzania. The study used a mixture of qualitative and quantitative interview methods to examine local perceptions of malaria and malaria treatment practices. Although the local term homa ya malaria or malaria fever appeared on the surface to correspond closely with the biomedical term malaria, significant and often subtle differences were found between the two terms. Of perhaps greatest importance, common consequences of malaria in endemic areas such as cerebral malaria in young children, severe anaemia and malaria in pregnancy were not connected with homa ya malaria by many people. A set of guidelines are described that were used to determine how best to promote acceptance and use of insecticide-impregnated mosquito nets, given these results. It is demonstrated that the position of the term used to denote malaria in the local taxonomy of febrile illnesses has important implications for the design of health education interventions. PMID- 8730912 TI - The impact on unemployment of an intervention to increase recognition of previously untreated anxiety among primary care physicians. AB - Anxiety is a common, though often unrecognized, problem in primary care settings. This study examines the effect on employment of an intervention designed to attune primary care physicians to previously unrecognized and untreated anxiety. Primary care physicians in a mixed-model health maintenance organization (HMO) were randomized by practice site to groups with (intervention) and without (usual care) intensive one-on-one education about anxiety and periodic feedback about their patients with anxiety. All persons 21-65 years of age presenting to the offices of these primary care providers were screened for anxiety with the SCL-90 R on two occasions. Those meeting the SCL-90-R cutpoints for anxiety and whose medical records provided no evidence of recognition or treatment for a mental health condition within the last 6 months were eligible for the study (n = 637). Of these, 573 (90%) completed two follow-up assessments. The present study evaluates the impact of the intervention aimed at the primary care physicians on the labor force participation rate of the persons with anxiety after 5 months of follow-up. The study also evaluates the impact of the intervention on hours of work and the presence of days spent in bed among the persons with anxiety working at the baseline interview and after 5 months. At baseline, the patients of intervention and usual care physicians with previously unrecognized and untreated anxiety did not differ in labor force participation rates. At the conclusion of the study, the patients of the intervention group physicians had significantly lower rates of labor force participation than those of the usual care group physicians. Among those working both at the beginning and conclusion of the study, the intervention had no impact on hours of work or the presence of days spent in bed. We conclude that attuning physicians may reduce labor force participation rates. PMID- 8730913 TI - Coping with additional stresses: comparative study of healthy and cancer patient new immigrants. AB - The adjustment and psychological distress of 166 cancer patients, who are new immigrants from the former Soviet Union, was assessed and compared to that of 288 healthy new immigrants from the the former Soviet Union. The healthy new immigrants had many adjustment problems and their psychological distress was fairly high. The cancer patients reported extremely severe psychological distress. In the healthy immigrants, age contributed to distress while family support had significant protective effects especially in the male immigrants. In the patients, these differences were even more extreme with family support being protective in the male group but not in the female group. Intrusiveness (IES) seems to be maladaptive adding to distress. The results clearly indicate that additional stresses, such as immigration, make cancer patients more vulnerable. The results also suggest possible interventions, especially those that will help to decrease intrusiveness. PMID- 8730914 TI - Age and health status in a patient satisfaction survey. AB - A picture of patients' satisfaction with interpersonal aspects of hospital-based care (including out-patient and accident and emergency services) was obtained from a postal survey of the general population of Lothian Region in south-east Scotland. Results were broadly in agreement with other national surveys and emphasized the high importance patients attach to being encouraged to ask questions about their treatment, and having their choices explained, including the right to a second opinion. Dissatisfaction decreased markedly with age, and also showed a weaker but significant association with measures of psychosocial health and pain. Social class was associated with feeling patronized or ignored by doctors. The opposite directions of the associations of satisfaction with older age and poorer health respectively, and the necessary correlation between age and health, imply that the effects of health status and age on satisfaction should be estimated from a multivariate model. PMID- 8730915 TI - A triple burden for health sector reform: 'post'-conflict rehabilitation in Uganda. AB - While conflict continues to threaten health development in many countries, relative peace has been secured in others. The transition from war to peace carries important political and economic opportunities for the reappraisal of social policy in general, and of health policy in particular. The health systems of countries recovering from prolonged periods of conflict often carry a double burden: the inheritance of an inappropriate and unaffordable health system developed in the pre-conflict era, and the particular, long-term effects of conflict on health and health services. This paper reports on the particular policies designed to rehabilitate the Ugandan health system, and argues that they exacerbated, rather than alleviated, the health crisis inherited in 1986. In this way they posed a third burden. By analyzing the context and process of policy formulation in the immediate post-conflict period, it explores the rationale which lay behind the adoption of these policies and identifies potential strategies for strengthening policy development in these unstable, resource-poor and health-deprived situations. PMID- 8730916 TI - Pipedreams. PMID- 8730917 TI - Distribution of remikiren, a potent orally active inhibitor of human renin, in laboratory animals. AB - 1. Whole-body autoradiography was used to compare the distribution of remikiren in the squirrel monkey, in which the compound is a potent inhibitor of renin, with the rat and the guinea-pig in which it is less active. 2. Following intravenous administration, drug-related material was rapidly and extensively taken up by the tissues of all three species. Consistent with rapid biliary elimination, high levels of radioactivity were found in the bile duct/gall bladder/intestinal contents. Of the remaining organs, the kidney consistently showed the highest concentrations of drug-related material. 3. Radio-hplc analysis of the kidney samples demonstrated that the majority of the retained material was present as intact remikiren, even at 24 h after administration. A similar degree of retention by the kidney was also found after oral dosing. 4. Uptake of remikiren by the kidney may act as a reservoir for the drug, resulting in the prolonged duration of pharmacological activity, which, despite the high plasma clearance of the drug, has previously been observed in primates. PMID- 8730918 TI - 1H and 19F-nmr spectroscopic studies on the metabolism and urinary excretion of mono- and disubstituted phenols in the rat. AB - 1. 1H and 19F-nmr spectroscopy was used to investigate quantitatively the urinary excretion of the metabolites of 15 substituted phenols in the rat. The compounds studied were: 2-, 3-, and 4-fluorophenols; 2-, 3-, and 4-trifluoromethylphenol; 2,4-, 2,6- and 3,4-difluorophenol; 2-fluoro-5-trifluoromethylphenol, 3-fluoro-5 trifluoromethylphenol, 2-trifluoromethyl-4-fluorophenol; 3-chloro-4-fluorophenol, 3-fluoro-4-chlorophenol, and 3-methyl-4-fluorophenol. All compounds were dosed to the Sprague-Dawley rat (10 mg/kg i.p.) and urine was collected over the periods 0 8, 8-24 and 24-48 h post-dosing and analyzed using nmr spectroscopy. 2. The compounds were excreted in the urine mainly as glucuronide or sulphate conjugates or as the unchanged parent compound. There was considerable variation in the urinary excretion of the compounds over 48 h ranging from 22.1 to 93.6% of the dose. There was no apparent relationship between the molecular weight of compounds or their metabolites and the percentage molar recovery of each in the urine. 3. Ortho-substituted phenols in general showed a greater propensity for glucuronidation than did either meta- or para-substituted compounds, irrespective of the substituent group. The molar glucuronide-to-sulphate ratio for ortho substituted compounds was found to be 2.2 +/- 0.9 whereas the ratio for both meta and para-substituted compounds was 0.8 +/- 0.2 (p < 0.0001). 4. There were characteristic substituent effects of phenolic glucuronidation or sulphation on the 19F-nmr chemical shifts for both F- and CF3-substituted phemols and these substituent effects were a useful aid to metabolite signal assignment. 5. These studies show that nmr spectroscopy provides a rapid and convenient approach to the construction of metabolic databases of simple xenobiotics for the investigation of structure-metabolism relationships. PMID- 8730919 TI - Inhibition of cyclosporine and tetrahydrocannabinol metabolism by cannabidiol in mouse and human microsomes. AB - 1. The in vitro and in vivo effects of cannabidiol on mouse and human liver microsomal metabolism of the immunosuppressive drug cyclosporine and the psychoactive compound tetrahydrocannabinol have been examined. 2. Preincubation of mouse or human liver microsomes with cannabidiol decreased the formation of all detectable cyclosporine metabolites by 73-89%. 3. In vivo cannabidiol treatment of mouse similarly decreased the formation of all detectable cyclosporine metabolites by 60-86%. 4. Preincubation of human liver microsomes with cannabidiol selectively decreased the formation of tetrahydrocannabinol metabolites catalyzed by cytochrome P4503A by 60% but had no effect on P4502C9 catalyzed metabolites. 5. Cannabidiol has the potential to clinically affect cyclosporine metabolism which may result in increased cyclosporine blood levels and an increase in its toxic side effects, and likewise may also affect tetrahydrocannabinol and its metabolite levels in man. PMID- 8730920 TI - Excretion and metabolism of remikiren, a potent orally active inhibitor of primate renin. AB - 1. Following intravenous administration of 14C-remikiren to the male rat, 78% of the administered radioactivity was recovered in faeces, indicating high biliary elimination. Of the 25 +/- 0.1% of the dose recovered in urine, the majority (16.5% of dose) was intact drug. 2. After oral administration to the male rat the urinary recovery was markedly reduced (8.5 +/- 2.0% of dose), and virtually all of the material was excreted as an inactive hydrolysis product. Intact drug was non-detectable, suggesting extensive first-pass metabolism. 3. Perfusion of isolated rat liver confirmed high biliary elimination, coupled with extensive metabolism. Although intact remikiren was the major component in bile (20% of the 'dose'), the majority of the radioactivity was recovered as a series of mono- and di-hydroxylated metabolites. 4. When screened against human renin, only one of the metabolites in bile and urine (mono-hydroxylated in the t-butyl side chain, and synthesized as Ro 44-0444) showed comparable activity to remikiren. The remaining ten metabolites tested were at least one order of magnitude less active than the parent drug. 5. In comparative in vitro studies Ro 44-0444 was formed by rat, but not human or cynomolgus monkey, liver microsomes. The primate microsomes also produced more of the remaining mono- and di-hydroxy products, suggesting that metabolites make little contribution to the oral activity of remikiren which is observed in these species in vivo. PMID- 8730921 TI - Induction of cytochrome P450 isoenzymes in cultured precision-cut rat and human liver slices. AB - 1. The effect of some xenobiotics on levels of selected cytochrome P450 (CYP) isoenzymes determined by Western immunoblotting and associated enzyme activities has been studied in 72-h cultured rat and human precision-cut liver slices. 2. In cultured rat liver slices, 0.5 mM sodium phenobarbitone (PB), 25 microM beta naphthoflavone (BNF), and 20 micrograms/ml Aroclor 1254 (ARO) induced mixed function oxidase enzyme activities. Western immunoblotting of liver slice microsomes was performed with antibodies to rat CYP1A2, 2B1/2 and 3A. Compared with 72-h control (dimethyl sulphoxide only treated) rat liver slice microsomes, PB induced CYP2B1/2 and 3A, BNF induced CYP1A2, and ARO induced CYP1A2, 2B1/2, and 3A. 3. The peroxisome proliferators methylclofenapate (MCP), ciprofibrate (CIP) and Wy-14,643 (WY) induced palmitoyl-CoA oxidation in 72-h cultured rat liver slices. Compared with 72-h control rat liver slice microsomes, MCP, CIP, and WY all induced levels of CYP4A. 4. In cultured human liver slices, 20 micrograms/ml ARO, but not 0.5 mM MCP, induced 7-ethoxyresorufin O-deethylase activity. Neither ARO nor MCP had any effect on homogenate palmitoyl-CoA oxidation and microsomal lauric acid 11- and 12-hydroxylase activities. Compared with 72-h control human liver slice microsomes, ARO induced CYP1A2, and MCP appeared to induce CYP4A. Further studies would be required to confirm that CYP4A isoenzymes could be induced by xenobiotics in human liver slices. 5. These results demonstrate that cultured liver slices may be used in evaluating the effect of xenobiotics on both rat and human CYP isoenzymes. PMID- 8730922 TI - Nutritional condition affects the disposition kinetics of albendazole in cattle. AB - 1. The influence of nutritional status on the plasma and abomasal fluid disposition kinetics of albendazole (ABZ) and its metabolites, albendazole sulphoxide (ABZSO) and albendazole sulphone (ABZSO2), has been investigated in the calf. 2. Free fatty acid (FFA) and beta-hydroxybutyrate (beta-OHB) serum concentrations were significantly higher in the feed-restricted (poor nutritional status) compared with control calf (optimal nutritional status). 3. ABZ parent drug was not detected in plasma at any time post-treatment and ABZSO and ABZSO2 were the metabolites detected in plasma. Both metabolites were rapidly depleted from the bloodstream. ABZ and its metabolites were recovered in abomasal fluid from 0.25 up to either 48 (ABZ) or 120h (ABZSO and ABZSO2) post-treatment. 4. The plasma disposition kinetics of both ABZ metabolites was significantly changed in the feed-restricted compared with control calf. ABZSO and ABZSO2 plasma area under the curves (AUCs) were significantly higher in the restricted animal. These enhanced AUCs correlated with significantly longer plasma half-lives (T1/2el) and mean residence times (MRTs) for these metabolites. 5. The delayed elimination of ABZ metabolites from the bloodstream correlated with the higher concentration of these molecules recovered in the abomasal fluid of the calves subjected to a dietary restriction. 6. The changes observed on disposition kinetics may reflect an impairment on the hepatic metabolism and clearance of ABZ as a consequence of FFA mobilization from adipose tissue and overproduction of ketone bodies in the liver. PMID- 8730923 TI - Identification and antimicrobial activity of urinary metabolites of a rifamycin derivative in dog. AB - 1. Three metabolites of the antimicrobial agent 3'-hydroxy-5'-(4-isobutyl-1 piperazinyl)benzoxazinorifamycin (KRM-1648) were isolated from dog urine obtained after administration of a single oral dose. These metabolites of KRM-1648 were identified by mass spectrometry and 1H and 13C-nmr spectrometry. 2. Three metabolites of KRM-1648 were identified as 25-deacetyl KRM-1648, 30-hydroxy KRM 1648 and 25-deacetyl-30-hydroxy KRM-1648. 3. The antimicrobial activities of 25 deacetyl KRM-1648 were comparable with those of the parent compound, whereas 30 hydroxy KRM-1648 was equipotent and 2-8-fold less active than the parent compound against bacteria and mycobacteria, respectively. PMID- 8730924 TI - Pharmacokinetics of remikiren, a potent orally active inhibitor of human renin, in rat, dog and primates. AB - 1. An hplc method with fluorescence derivatization was developed for the quantification of remikiren in plasma (limit of quantification 2 ng/ml). This was used to determine the pharmacokinetics in various species of primate, in which the compound is a potent inhibitor of renin, as well as in the rat and dog in which it is less active. 2. After intravenous administration the mean residence time was < or = 1.5 h in all species, and the plasma clearance approached the corresponding hepatic blood flows. 3. Studies in the bile-duct cannulated rat and dog demonstrated that the high clearance was due to a combination of rapid metabolism, plus biliary and renal excretion of intact drug. 4. Consistent with the high hepatic clearance, oral bioavailability was low ( < or = 6%) in each species. However, all of the species tested absorbed a small proportion of an oral dose extremely rapidly, to give peak concentrations generally within 5 min of administration. 5. 'Simultaneous' collection of blood samples from the hepatic portal vein and aorta of rat confirmed that shortly after oral dosing the intact drug did cross the liver; however, the later collections contained predominantly more polar metabolites. 6. The rapid absorption of intact remikiren is consistent with the transient blockade of plasma renin activity, previously observed in primates after oral administration. However, the high clearance appears inconsistent with the subsequent prolonged decrease in blood pressure, suggesting that the latter effect is mediated through a 'tissue' compartment. PMID- 8730925 TI - Metabolism of illudin S, a toxic substance of Lampteromyces japonicus: urinary excretion of mercapturic acids in rat. AB - 1. The urinary excretion of the mercapturic acids of illudin S after oral administration to rat has been studied. 2. From lc-ms/ms analysis of methanolic extracts of lyophilized rat urine, stereo-isomeric mercapturic acids were detected. 3. The mercapturic acids excreted 3 days following administration amounted to approximately 0.39-0.73% of the administered dose. 4. In vitro glutathione conjugation of illudin S by subcellular fractions was also examined. 5. No significant increases in the formation of glutathione adducts were observed in any subcellular fractions examined. PMID- 8730926 TI - Metabolism of tamsulosin in rat and dog. AB - 1. The metabolism of tamsulosin hydrochloride (TMS), a potent alpha 1 adrenoceptor blocking agent, was studied after a single oral administration to rat and dog. 2. Eleven metabolites (1, 2, 3, 4 and their glucuronides, sulphates of 1 and 3, and A-1) were identified from the urine and bile of rat and dog administered TMS. 3. Unchanged drug and metabolites in urine and bile were quantified in rat and dog dosed with 14C-TMS(1 mg/kg). In rat the main metabolic routes were de-ethylation of the o-ethoxyphenoxy moiety, demethylation of the methoxybenzenesulphonamide moiety, and conjugation of the resultant metabolites by glucuronic acid and sulphuric acid. In dog the main pathways were de ethylation of the ethoxyphenoxy moiety, conjugation of the de-ethylated product by sulphuric acid, and oxidative deamination of the side chain. 4. The organ responsible for the metabolism of TMS in rat was estimated using 9000g supernatants of liver, kidney, small and large intestine homogenate and plasma. The drug was rapidly metabolized in liver but hardly metabolized in the other organs or plasma. PMID- 8730927 TI - In vitro binding of synaptic vesicles to the synaptic plasma membrane: lack of effect of beta-bungarotoxin. AB - To help characterize the mechanisms of neurotransmitter release, and the role of the specific neurotoxin beta-bungarotoxin in inhibiting release, the interaction of synaptic vesicles with the synaptic plasma membrane was investigated using two in vitro systems. Binding of radiolabeled synaptic vesicles to immobilized synaptic plasma membrane was specific, protein-dependent, and modulated by phosphorylation of membrane proteins. Stimulation of phosphorylation by phorbol ester increased binding, and reduction of phosphorylation by alkaline phosphatase or staurosporine reduced binding. beta-Bungarotoxin did not alter basal binding of synaptic vesicles to synaptic plasma membrane, nor did it affect the increase in binding induced by phorbol esters. Under conditions which stimulate acetylcholine release from synaptosomes, both phorbol ester and 4-aminopyridine caused an increase in attachment of the synaptic vesicle marker protein synaptophysin to the synaptic plasma membrane. beta-Bungarotoxin did not alter the change in localization of synaptophysin induced by either drug, under conditions in which it inhibits ACh release induced by 4-aminopyridine. It is concluded that beta-bungarotoxin inhibition probably does not occur at the level of the interaction of the synaptic vesicle and the synaptic plasma membrane, but occurs at an earlier stage in the neurotransmission process. PMID- 8730928 TI - Dinophysistoxin-2: the predominant diarrhoetic shellfish toxin in Ireland. AB - Diarrhoetic shellfish poisoning (DSP) in Europe is due mainly to the presence of the dinoflagellate toxin, okadaic acid (OA). However, analysis of cultivated mussels (Mytilus edulis) from southwest Ireland revealed that an isomer of OA, dinophysistoxin-2, was the major toxin present during DSP episodes. Using fluorimetric HPLC, following derivatisation with 9-anthryldiazomethane, both OA and DTX-2 were found in shellfish during a prolonged toxic episode in 1991. However, examination of similar mussel cultivation locations in 1994 showed that DTX-2 was even more predominant. During this DSP period, OA levels were less than 0.7 microgram/g, whereas maximum DTX-2 levels of 6.3 micrograms/g hepatopancreas were recorded. This toxicity in shellfish occurred soon after high cell counts of Dinophysis acuta were observed. As well as large seasonal variability in toxin levels in rope cultured mussels, substantial variations were also observed, both horizontally and vertically, within the water column. PMID- 8730929 TI - Larvotoxic extracts of the hard coral Goniopora tenuidens: allelochemicals that limit settlement of potential competitors? AB - Aqueous extracts of Goniopora tenuidens inhibited larval metamorphosis and post metamorphic growth of the hard coral Pocillopora damicornis. The extracts killed swimming larvae of four scleractinian corals and modified the behaviour of larvae of P. damicornis at concentrations as low as 3.9 micrograms ml-1. This toxicity may indicate the presence of allelochemicals that limit the settlement of potential competitors. PMID- 8730930 TI - Antibody-mediated neutralization and binding-reversal studies on alpha neurotoxins from Micrurus nigrocinctus nigrocinctus (coral snake) venom. AB - An ELISA based, non-radioactive acetylcholine receptor (AchR) binding assay was used to detect the alpha-neurotoxins present in Micrurus nigrocinctus nigrocinctus venom. Sera from horses hyperimmunized against M. nigrocinctus venom contain antibodies which inhibit the binding of M. n. nigrocinctus alpha neurotoxins to AchR and reverse the binding of toxins already complexed with the receptor. This result supports the importance of using antivenom therapeutically in M. n. nigrocinctus envenomations even after the onset of neurological symptoms. M. nigrocinctus antivenoms cross-reacted in an ELISA with several elapid alpha-neurotoxins and inhibited the binding of Bungarus multicinctus alpha bungarotoxin and Naja naja oxiana neurotoxin II to AchR in vitro, suggesting the presence of short-chain and long-chain alpha-neurotoxins in M. nigrocinctus venom. In vivo neutralization experiments with M. nigrocinctus antivenom demonstrate that M. nigrocinctus venom contains short-chain alpha-neurotoxin(s) which share common neutralizing epitope(s) with Naja naja oxiana neurotoxin II. PMID- 8730931 TI - Detection of okadaic acid esters in the hexane extracts of Spanish mussels. AB - Two types of low polar derivatives of OA and dinophysitoxins have been reported in shellfish or in phytoplankton: 7-0-acyl esters containing a fatty acyl group attached through the 7-OH group and diol esters in which the carboxylic group of the toxins has been esterified. These compounds cannot be directly detected by liquid chromatography and fluorimetric detection as 9-anthryldiazomethane derivatives, owing in the first case to their low polarity and high molecular weight, and in the second case because they have the carboxylic group esterified. All of them must be hydrolysed before derivatization to be detected as Adam derivatives of the corresponding non-acylated toxins. In the Lee procedure, after extraction of the shellfish digestive glands with 80% methanol, a liquid-liquid partition with a non-polar solvent such as hexane is carried out in order to remove non-polar lipids. The presence of non-polar toxins was investigated in Spanish mussels and confirmed in the hexane layer, usually discarded in conventional extraction procedures, by analysis of the alkaline hydrolysis products. A preferred solubilization of these toxins in a non-polar solvent like hexane is reported. The inclusion of a hydrolytic step of the hexane extract in the general procedure is suggested in order to monitor the contribution of non polar diarrhoetic shellfish poisons (DSPs) to the total DSP shellfish toxicity. This is the first report of DSPs other than OA and DTX2 in Spanish mussels. PMID- 8730932 TI - Interleukin-6 release following scorpion sting in children. AB - Interleukin-6 levels were measured in the serum of ten children following severe scorpion envenomation. Measurements were taken on arrival, at the emergency room, and 12 and 24 hr after arrival. Interleukin-6 was markedly elevated in the serum of eight out of ten children on arrival. Interleukin-6 levels gradually decreased toward normal values on 12 and 24 hr measurements, but remained above control levels on all measurements. These results imply that signs and symptoms following scorpion envenomation may in part be explained by release of cytokines. Human and experimental animal studies are required in order to verify the assumption that interleukin-6 and other cytokines are involved in the pathogenesis of scorpion envenomation. PMID- 8730933 TI - Schizophrenia, CATCH 22 and FISH. PMID- 8730934 TI - Accident neurosis revisited. PMID- 8730935 TI - Are SSRIs a cost-effective alternative to tricyclics? AB - BACKGROUND: Selective serotonin reuptake inhibitors (SSRIs) are more expensive than tricyclics. Reports have suggested that SSRIs are cost-effective because they are better tolerated and safer in overdose. METHOD: A systematic review of all randomised controlled trials (RCTs), meta-analyses, and cost-effectiveness studies comparing SSRIs and tricyclic antidepressants (TCAs). RESULTS: None of the RCTs provided an economic analysis and there were methodological problems in the majority which would preclude this approach. Meta-analyses suggest that clinical efficacy is equivalent but slightly fewer patients prescribed SSRIs drop out of RCTs. Cost-effectiveness studies have been based on crude 'modelling' approaches and over-estimate the difference in attrition rates and the cost of treatment failure. It appears impossible to evaluate the economic aspects of suicide because of its rarity. CONCLUSIONS: There is no evidence to suggest that SSRIs are more cost-effective than TCAs. The debate will only be concluded when a prospective cost-effectiveness study is done in the setting of a large primary care based RCT. PMID- 8730936 TI - The Camden schizophrenia surveys. I. The psychiatric, behavioural and social characteristics of the severely mentally ill in an inner London health district. AB - BACKGROUND: The development of appropriate community care requires knowledge of the characteristics of the severely mentally ill. METHOD: All patients with a broad diagnosis of schizophrenia (n = 528) were identified via key informants within an inner London health district; Feighner and DSM-III-R criteria for schizophrenia were applied. Clinical and social status, psychopathology (Manchester Scale) and problem behaviours (Social Behaviour Schedule) were assessed in a 4:5 representative sample. RESULTS: Fifty-seven per cent had experienced at least one compulsory admission. Forty-five per cent (excluding long-stay in-patients) had marked positive psychotic symptoms; social isolation and lack of daytime activity was more common than among comparable populations elsewhere. One hundred and eleven (25%) of the non-hospitalised patients were not in contact with specialist services. CONCLUSIONS: Difficulties in establishing and maintaining a therapeutic alliance between patients and professionals present a challenge to services in inner London. Many social and occupational needs were not being met by existing community provision. PMID- 8730937 TI - The Camden schizophrenia surveys. II. High prevalence of schizophrenia in an inner London borough and its relationship to socio-demographic factors. AB - BACKGROUND: Investigation of the geographical distribution of schizophrenia and its relationship to socio-demographic factors is useful for planning services. METHOD: Individuals with schizophrenia (n = 980) were identified by key informants within an inner London borough and point prevalence calculated for broad, Feighner and DSM-III-R schizophrenia. The distribution of cases was tested for significant variation using the Poisson process model. Regression models using the Jarman-8 score and its component variables were tested for their ability to predict the prevalence of schizophrenia. RESULTS: A high point prevalence of schizophrenia (5.3 per 1000 resident population) was demonstrated. Case distribution showed a marked and significant variation associated with socio demographic factors. The prediction of prevalence was more accurate for broad than for narrower definitions of schizophrenia; unemployment rate performed best. CONCLUSIONS: Unemployment rates and Jarman-8 scores may provide crude estimates for resource allocation in planning mental health services, highlighting the need for additional services in deprived inner city areas. PMID- 8730938 TI - Patients with schizophrenia remember that an event has occurred, but not when. AB - BACKGROUND: The context memory deficit hypothesis of schizophrenia postulates that the long-term deficit associated with this disorder is related to a memory impairment for contextual information. METHOD: To test this hypothesis, memory for temporal context was assessed in 33 patients with schizophrenia and 33 normal subjects, using a recency discrimination task. RESULTS: Whereas patients were able to recall and recognise target items, they were unable to recognise from among the target items those which had been most recently learned. CONCLUSIONS: Schizophrenia is associated with a temporal context memory deficit. PMID- 8730939 TI - The dimensions of schizophrenia phenomenology. Not one or two, at least three, perhaps four. AB - BACKGROUND: This confirmatory investigation examined the underlying structure of schizophrenia phenomenology through examination of the fit of several prominent dimensional models to observed symptom data. METHOD: Confirmatory factor analysis was conducted on a correlation matrix of schizophrenia signs and symptoms derived from case history ratings of 192 individuals with schizophrenia who were the subjects in the major twin studies of schizophrenia. RESULTS: Schizophrenia phenomenology appears best described by four underlying factors, namely negative symptoms, premorbid social adjustment deficits, reality distortion, and disorganisation. Of interest, the premorbid deficit dimension was directly associated with negative symptoms and disorganisation, but was inversely associated with reality distortion. CONCLUSIONS: These data clearly support the multidimensionality of schizophrenia phenomenology and provide objective support for a four-factor model over other models. This four-factor model may be useful in organising existing and future data concerning the genetic, neurobiological, neurological, and psychosocial features of schizophrenia. PMID- 8730940 TI - Public education for community care. A new approach. AB - BACKGROUND: The findings from a controlled study of the effect of a public education campaign on community attitudes to mentally ill people are presented. METHOD: A census of neighbours' attitudes toward mental illness was conducted in two areas before the opening of supported houses for the mentally ill. In one area an educational campaign was conducted. The attitude survey was then repeated in both areas and patients' social contact with neighbours was recorded. RESULTS: Respondents exposed to the didactic component of the campaign showed only a small increase in knowledge about mental illness but there was a lessening of fearful and rejecting attitudes in the experimental area and not in the control area. Neighbours in the experimental area were more likely to make social contact with both staff and patients. It was social contact which was directly associated with improved attitudes rather than education per se. Patients in the experimental area made contact and even friendships with neighbours whereas those in the control area did not. CONCLUSIONS: The public education campaign did not lead to significant change in neighbours' knowledge of mental illness. However, their attitudes improved and patients' social integration was enhanced. PMID- 8730941 TI - Residential care in hospital and in the community--quality of care and quality of life. AB - BACKGROUND: The reduction of beds in long-stay hospitals has led to concerns over the quality of care offered to the remaining residents as well as that provided in the community. This study seeks to compare the quality of care and quality of life (reported satisfaction) from residents in both types of setting. METHOD: A cross-sectional comparison was made of community residential homes and hospital wards drawn randomly from lists provided by local authorities in the outer London area. Samples were drawn from all the main provider types (local authority, housing association, private and joint NHS/voluntary sector). Measures were taken of the quality of the physical environment, staff and resident characteristics, external management arrangements and internal management regimes, resident satisfaction and staff stress. Direct observations were also made of the amount and quality of staff-resident interactions. RESULTS: In general, the most disabled residents were found to be still living in hospital in the Worst conditions and receiving the poorest quality of care. Although there were some problems with missing data, hospital residents also seemed most dissatisfied with their living situation. There were few differences between community providers regarding either the quality of care provided or the levels of reported satisfaction. Quality of care in the community homes seemed to be much more determined by the personality and orientation of project leaders. CONCLUSIONS: Purchasers and providers still need to give attention to the problems of selectively discharging the most able residents to the community, leaving the most disabled being looked after in progressively deteriorating conditions. All residential providers need to review their internal management practices and try to ensure that residents are offered, as far as possible, the opportunity to make basic choices about where and how they will live. Staff training and quality assurance practices need to be reviewed in order to improve the direct quality of care offered to the most disabled individuals. PMID- 8730942 TI - A controlled trial of dothiepin and placebo in treating benzodiazepine withdrawal symptoms. AB - BACKGROUND: The possibility that treatment with tricyclic antidepressants, in the form of dothiepin, might attenuate benzodiazepine withdrawal symptoms was investigated in a double-blind trial. METHOD: Eighty-seven non-depressed psychiatric out-patients with putative normal dose benzodiazepine dependence had their benzodiazepines reduced in stepwise amounts of 20% of the original dose for eight weeks. The patients were randomised to receive dothiepin (with dosage increasing to 150 mg/day) or placebo as an aid to withdrawal before benzodiazepine reduction and these drugs were taken for four further weeks before being stopped. RESULTS: Fewer patients entered and completed the study than expected and a Type II error was possible in the results. Although there was some evidence of withdrawal symptoms being less marked in those patients allocated to dothiepin this was independent of any antidepressant effect as depression scores were lower in the placebo group in the early phase of withdrawal (P < 0.01). Of those completing the study, greater satisfaction (P = 0.03) was recorded by those who had received dothiepin; no other differences reached statistical significance. CONCLUSIONS: Dothiepin (and by implication other tricyclic antidepressants) might have some value in reducing benzodiazepine withdrawal symptoms but does not aid drug withdrawal. PMID- 8730943 TI - The Galway Study of Panic Disorder. III. Outcome at 5 to 6 years. AB - BACKGROUND: The aim was to evaluate long-term outcome of DSM-III-R panic disorder at a mean of 5.3 years following a controlled trial of treatment that included antidepressants and behavioural counselling. METHOD: Sixty-eight (86%) subjects were evaluated by lengthy research interview. RESULTS: Thirty-four per cent recovered and remained well, 46% were minimally impaired and 20% had persistent panic disorder of whom half remained significantly impaired. Anxious-fearful personality dysfunction was the most important predictor of poor outcome, followed by poor clinical status at discharge and inability at baseline to recall vividly the initial panic attack. Those who dropped out from the original trial did badly. CONCLUSIONS: Complete recovery can occur even after many years of severe illness in a large minority of subjects who receive both antidepressants and behavioural counselling in the acute stage of treatment. The comparative prognostic value of personality, severity and chronicity need to be more fully addressed in future studies. PMID- 8730944 TI - A typology of sub-clinical senescent cognitive disorder. AB - BACKGROUND: Cognitive impairment without dementia is commonly observed in ageing populations. The present study aims to describe types of impairment and evolution over a one-year period. METHOD: Three hundred and ninety-seven normal French elderly persons demonstrating recent, observable change in cognitive performance were examined annually using a computerised cognitive examination. RESULTS: Five subtypes were differentiated by cluster analysis. Two of the groups were predicted by logistic regression to be at high risk of senile dementia. Of 16 incident cases of senile dementia diagnosed in the following year, 13 were found to have derived from these two groups. The typology was also found to be useful in the description of age-associated memory impairment. CONCLUSIONS: Subclinical cognitive impairment was found to not constitute a unitary phenomenon and heterogeneous subgroups could be differentiated. The concept of 'normality' in elderly cohorts is reconsidered in the light of these findings. PMID- 8730945 TI - A magnetic resonance imaging study of white matter lesions in depression and Alzheimer's disease. AB - BACKGROUND: White matter changes, as revealed by magnetic resonance imaging (MRI), may occur in depression and Alzheimer's disease. METHOD: T2-weighted MRI scans were performed in 39 control subjects, 61 subjects with NINCDS/ADRDA Alzheimer's disease and 60 subjects with DSM-III-R major depression. Deep white matter lesions (DWML) and periventricular lesions (PVL) were rated on a standard 0-3 scale by two radiologists blind to clinical diagnosis. RESULTS: After controlling for differences in vascular risk factors and current blood pressure, DWML were significantly more common in depressed subjects and PVL in Alzheimer's disease subjects compared to controls. DWML were most common in those presenting in late life with their first ever depression and 50% of such subjects had severe (grade 3) DWML. CONCLUSION: An association between DWML and depression and PVL and Alzheimer's disease is supported. The increase with DWML that occurs with ageing may predispose some elderly subjects to depression. PMID- 8730946 TI - The diagnosis and prevalence of hyperactivity in Chinese schoolboys. AB - BACKGROUND: This study was undertaken to examine the validity of different diagnostic definitions of hyperactivity in a Chinese population. Estimates of the prevalence of hyperactivity were made according to these different diagnostic definitions. METHOD: In a two-stage epidemiological study of hyperactivity in Hong Kong, 3069 Chinese schoolboys were screened by questionnaires; and a stratified sample of 611 of them entered a second stage for more detailed diagnostic assessment. RESULTS: Children with hyperkinetic disorder (ICD-10) or ADDH (DSM-III) both displayed significant hyperactive symptoms, but with somewhat different external correlates; hyperkinetic disorder tended to show more neurodevelopmental impairments, ADDH more cognitive and educational difficulties. These findings raise the possibility of heterogeneity in the disorders present with hyperactivity. The DSM-III-R category of ADHD was more common, and those extra cases, that did not overlap with ADDH or hyperkinetic disorder, included children with no obvious behavioural, cognitive or neurodevelopmental impairments. Hence ADHD may be an over-inclusive category. Prevalence rates for hyperkinetic disorder, ADDH and ADHD were respectively 0.78%, 6.1% and 8.9%. CONCLUSIONS: A disorder of hyperactivity does exist in the Chinese culture, displaying the same kinds of symptomatology and external correlates as in the West. The prevalence rates of hyperkinetic disorder and ADDH in Chinese schoolboys are on the low side when compared to those reported in Western studies. PMID- 8730947 TI - Confirmatory factor analysis of the Fear Questionnaire with social phobia patients. AB - BACKGROUND: The Fear Questionnaire is an internationally popular measure in anxiety disorder research, but its validity with social phobia patients has not been clearly demonstrated. METHOD: A confirmatory factor analysis was conducted using the Fear Questionnaire responses of 122 social phobia patients. RESULTS: The results indicated that the proposed three-factor model of the scale (agoraphobia, social phobia, and blood/injury dimensions) provided a good fit to the data. CONCLUSIONS: These data provide strong support for the validity of the scale and for the multidimensional view of fears proposed by Arrindell. PMID- 8730948 TI - Should amenorrhoea be necessary for the diagnosis of anorexia nervosa? Evidence from a Canadian community sample. AB - BACKGROUND: This study compares the characteristics of women with anorexia nervosa with those of women who have all the diagnostic features of that disorder except amenorrhoea. METHOD: The study uses data from a large community epidemiological survey of the mental health status of household residents in Ontario, Canada. A multi-stage stratified sampling design generated a sample of 4285 females aged 15-64. DSM-III-R diagnoses were made using the Composite International Diagnostic interview. RESULTS: Eighty-four out of 4285 female respondents met full or partial-syndrome criteria for anorexia nervosa. Comparison of these two groups revealed few statistically significant differences in terms of demographics, psychiatric comorbidity, family history or early experiences. CONCLUSIONS: Amenorrhoea did not discriminate between women with anorexia nervosa and women with all the features except amenorrhoea across a number of relevant variables. The authors question the utility of amenorrhoea as a diagnostic criterion. PMID- 8730949 TI - Abnormal electroretinogram in cocaine-dependent patients. Relationship to craving. AB - BACKGROUND: The retina contains high concentrations of dopamine. We previously reported that recently withdrawn cocaine-dependent patients had significantly reduced blue cone b wave amplitude responses on the electroretinogram (ERG). In this study we wished to examine if there was a relationship between cocaine craving and reduced blue cone b wave ERG responses. METHODS: A series of 14 recently withdrawn cocaine-dependent patients completed a 45-item cocaine craving questionnaire shortly before the ERG. RESULTS: Cocaine dependent patients who had a reduced ( < 0.5 microvolts) blue cone b wave ERG response had significantly higher cocaine craving scores than patients without a reduced blue cone ERG response. There was also a significant negative correlation between blue cone b wave amplitude responses and scores on the cocaine craving questionnaire. CONCLUSIONS: A reduced blue cone b wave ERG response may identify recently withdrawn cocaine-dependent patients with biological dysregulation and increased cocaine craving. PMID- 8730950 TI - The use of environmental strategies and psychotropic medication in the management of delirium. AB - BACKGROUND: This paper examines the pattern and frequency of implementation of environmental strategies and the use of psychotropic medication in the management of patients with delirium in an acute hospital setting. METHOD: The study involved 46 consecutive referrals to a consultation psychiatry service each of whom met ICD-10 criteria for delirium. Patients were subdivided into hyperactive, hypoactive and mixed subtypes of delirium and assessed regarding severity of delirium, the use of psychotropic medication prior to consultation and the implementation of environmental measures in their management. RESULTS: Mean age was 60.1 years. Thirty per cent of patients were of the hyperactive subtype, 24% hypoactive and 46% mixed. Psychotropic medication was given to 56.5% prior to consultation and this is significantly associated with severity of delirium and in particular, with hyperactive delirium subtype. Of eight environmental strategies only four were instituted in over 50% of the patients prior to consultation. The application of these strategies was associated with overall severity of delirium, agitation, mood lability and sleep-wake cycle disturbance. It was not significantly associated with severity of disorientation or with disturbed perception/thinking. CONCLUSION: Simple environmental strategies such as limiting changes in staff, minimising noise levels and involving relatives in re-orientation are frequently overlooked in the management of patients with delirium. Our study suggests that the implementation of environmental strategies occurs primarily in responses to behavioural challenges rather than to limit the core features of delirium. PMID- 8730951 TI - Successful treatment of tardive dystonia with clozapine and clonazepam. AB - BACKGROUND: Tardive dystonia is an uncommon complication of neuroleptic treatment which is frequently disabling and poorly responsive to treatment. METHOD: The case is reported of a 28-year-old patient with schizophrenia who developed severe, generalised tardive dystonia after five years of neuroleptic treatment. Stopping neuroleptic medication and treatment with tetrabenazine, an anticholinergic and a benzodiazepine were ineffective. Treatment with clozapine and then the novel combination of clozapine plus clonazepam was instituted. RESULTS: Treatment with clozapine alone brought about limited improvement. Addition of clonazepam resulted in virtually complete disappearance of all abnormal movements. This remission has been sustained for nearly two years. CONCLUSIONS: This report adds to two other cases suggesting that the combination of clozapine and clonazepam may be an effective treatment for tardive dystonia. PMID- 8730952 TI - Assessment of mental capacity: a dilemma. PMID- 8730953 TI - Paracetamol overdose. PMID- 8730954 TI - Drug-induced dysphoria. PMID- 8730955 TI - Low incidence of mania in northern Finland. PMID- 8730956 TI - Seasonality of birth in Western Australia. PMID- 8730957 TI - Presenile dementia in a Down's syndrome adult with an unbalanced 21/21 Robertsonian translocation. PMID- 8730958 TI - Preparation and characterization of polyethyl-2-cyanoacrylate nanocapsules containing antiepileptic drugs. AB - Biocompatible and biodegradable colloidal drug delivery systems can be obtained by means of in situ polymerization of alkylcyanoacrylate. In particular, nanocapsules of polyethylcyanoacrylate (PECA) were prepared by adding the monomer to an organic phase, consisting of Miglyol 812 and an organic solvent (ethanol, acetone or acetonitrile), and subsequently mixing the organic phase with an aqueous phase containing Pluronic F68 at different concentrations. The possible mechanism of formation and the influence of preparation conditions on the quality of nanocapsule formulations were investigated by freeze-fracture electron microscopy and laser light scattering using both the inverse Laplace transform and the standard cumulant analysis for data fitting. High-quality nanocapsule systems were obtained using an aprotic fully water-miscible organic solvent such as acetone. The presence of ethanol led to the formation of both nanospheres and nanocapsules. The concentrations of nonionic surfactant in the aqueous phase of monomer in the organic phase did not influence the kind of colloidal suspension obtained. The oil simply plays the role of monomer support. The diameter of PECA nanoparticles (nanospheres and nanocapsules) ranged from 100 to 400 nm. Three antiepileptic drugs (Ethosuximide, 5,5-diphenyl hydantoin and carbamazepine) were entrapped in PECA nanocapsules. The loading capacity of PECA nanocapsules, prepared using acetone as organic solvent, varied from 1% to 11% (drug/dried material) as a function of the solubility (affinity) of the different drugs with the oil core. This parameter also influenced the release from PECA nanocapsules, which was slower for drugs with a higher affinity for Miglyol 812. By encapsulating the three antiepileptic drugs in the PECA nanocapsules, it was possible to achieve controlled drug release. The mechanism of drug release from PECA nanocapsules was mainly diffusion from the oil core through the intact polymer barrier. PMID- 8730959 TI - Surfaces modified with covalently-immobilized adhesive peptides affect fibroblast population motility. AB - Cell population motility and adhesion of rat skin fibroblasts were evaluated on aminophase glass modified with covalently-immobilized biologically active peptides, specifically, either arginine glycine-aspartic acid-serine (RGDS) or tyrosine-isoleucine-glycine-serine-arginine-glycine (YIGSRG). Fibroblast population motility was decreased and adhesion was increased on substrates modified with covalently immobilized RGDS peptide compared to substrates with the covalently immobilized non-adhesive peptides arginine-glycine-glutamic acid serine and arginine-aspartic acid-glycine-serine. Fibroblast motility was not significantly changed on substrates modified with covalently-immobilized YIGSRG peptide; however, fibroblast adhesion was decreased on that substrate. PMID- 8730960 TI - Cross-linking of dermal sheep collagen using a water-soluble carbodiimide. AB - A cross-linking method for collagen-based biomaterials was developed using the water-soluble carbodiimide 1-ethyl-3-(3-dimethyl aminopropyl)carbodiimide hydrochloride (EDC). Cross-linking using EDC involves the activation of carboxylic acid groups to give O-acylisourea groups, which form cross-links after reaction with free amine groups. Treatment of dermal sheep collagen (DSC) with EDC (E-DSC) resulted in materials with an increased shrinkage temperature (Ts) and a decreased free amine group content, showing that cross-linking occurred. Addition of N-hydroxysuccinimide to the EDC-containing cross-linking solution (E/N-DSC) increased the rate of cross-linking. Cross-linking increased the Ts of non-cross-linked DSC samples from 56 to 73 degrees C for E-DSC and to 86 degrees C for E/N-DSC samples, respectively. For both cross-linking methods a linear relation between the decrease in free amine group content and the increase in Ts was observed. The tensile strength and the high strain modulus of E/N-DSC samples decreased upon cross-linking from 18 to 15 MPa and from 26 to 16 MPa, respectively. The elongation at break of E/N-DSC increased upon cross-linking from 142 to 180%. PMID- 8730961 TI - Chondroitin-6-sulphate incorporated into collagen gels for the growth of human keratinocytes: the effect of cross-linking agents and diamines. AB - This study demonstrates the effect of the glycosaminoglycans, hyaluronic acid and chondroitin-6-sulphate (Ch6SO4), diamines and a carbodiimide cross-linking agent on the growth of human epidermal cells on collagen gels. Ch6SO4 incorporated into collagen gels stimulated cell growth rate, but the effect was found to be inconsistent. We found that approximately 50% of the incorporated Ch6SO4 in the gels leached out into the growth medium after the first 3 d in culture, and this is thought to lead to the inconsistent cell growth response. In order to minimize the elution of Ch6SO4 from the gels and thereby maximize its effect on the growth of the keratinocytes, 1-100 micrograms ml-1 Ch6SO4 was added in the medium. The results showed that Ch6SO4 at these concentrations in the medium did not stimulate the cell growth on either plain collagen gels or gels containing 20% Ch6SO4. As an alternative strategy, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide and diamines (putrescine or diaminohexane) were used to immobilize Ch6SO4 onto the collagen gels and to cross-link the gels. The cross-linking process partially prevented the elution of Ch6SO4 from the gels. Interestingly, only putrescine, not diaminohexane, promoted the growth of keratinocytes on the cross-linked plain collagen gels. We proposed to develop an artificial skin substitute containing putrescine as a growth factor for the human epidermal cells. PMID- 8730962 TI - Long-term compressive property durability of carbon fibre-reinforced polyetheretherketone composite in physiological saline. AB - In total hip arthroplasty, concerns such as corrosion and stress shielding associated with stiff metallic femoral components have led to the development of low stiffness advanced fibre-reinforced polymer (FRP) composite femoral components. Carbon fibre-reinforced polyetheretherketone (CF/PEEK) composite material is now one of the primary material systems being considered for composite hip stem development. As a hip stem, a composite material must be able to support a complex state of stress in the in vivo environment without failure. Considering the loading conditions of a hip stem (superimposed compression and bending), and the fact that FRP composites typically possess lower compressive than tensile strength, the compressive behaviour of FRP composites becomes very important for femoral component design. This paper presents an investigation of the long-term durability of 0 degree and 90 degrees compressive strengths of CF/PEEK composite following physiological saline saturation. 0 degree and 90 degrees compressive moduli and Poisson ratio (v12) properties are also reported. Samples were tested following conditioning in physiological saline at 37, 65 and 95 degrees C for time periods from 0 to 5000 h. Dry samples were tested as controls. Results show no significant loss in compressive property values of the saline-saturated or the dry control samples as a function of conditioning time or temperature. PMID- 8730963 TI - Evaluation of guided bone regeneration in rabbit tibia using bioresorbable and non-resorbable membranes. AB - The aim of this study was an evaluation of the possibility of bone regeneration in connection with implant placement, using a new bioresorbable membrane (Guidor Matrix Barrier) used previously in periodontal tissues regeneration. The study compared the bone regeneration obtained around Bonefit-ITI implants inserted in rabbit tibia using Guidor membranes, Gore-Tex membranes and in control sites. Microscopic analysis was performed after 6, 9 and 12 weeks. It was possible to see that the amount of bone around implants covered by Guidor and Gore-Tex membranes was roughly equivalent in all experimental sites at the 6-, 9- and 12 week time intervals. Also, all control sites healed completely. It could be concluded from this study that: (1) rabbit tibia cannot be recommended in research connected with guided bone regeneration, as all control sites healed in the same way as the test sites; (2) in no case did the presence or the degradation of the resorbable membrane prevent the formation of new bone; (3) no inflammatory reaction was present around the bioresorbable and the non-resorbable membranes; (4) bone formed in all cases on the outer surface of the non resorbable membranes; (5) large Guidor membrane fragments were present in the 6 week specimens, while in the 12-week specimens only small fragments were recognizable; (6) Guidor membranes can be used in guided bone regeneration. PMID- 8730964 TI - Evaluation of soft tissue reactions at the interface of titanium limited contact dynamic compression plate implants with different surface treatments: an experimental sheep study. AB - Five types of limited contact dynamic compression plates (LC-DCPs) of pure titanium with different surface treatments and an electropolished stainless steel LC-DCP were tested. The surface roughness parameters and chemical surface conditions were determined and checked for probable surface contamination. After an implantation period of 3 months on long bones of sheep, the soft tissue adjacent to the plates was evaluated histomorphometrically. The difference in roughness parameters was statistically significant for most surface conditions. A correlation was found between the surface roughness of the implants and the thickness of the adjacent soft tissue layer. The thinnest soft tissue reaction layer with a good adhesion to the implant surface was observed for the titanium anodized plates with coarse surface. Smooth implants, in particular the electropolished stainless steel plates, induced statistically significant thicker soft tissue layers. Profilometer roughness measurements combined with scanning electron microscopy techniques were useful methods to characterize the surface morphology. PMID- 8730965 TI - Benign response to particles of diamond and SiC: bone chamber studies of new joint replacement coating materials in rabbits. AB - Wear particles from total joint replacements are thought to accelerate prosthetic loosening. Diamond coating may improve the smoothness and wear characteristics of the femoral head component of total hip replacements, and thus increase their longevity. The brittleness of a thin diamond coat may be overcome by using an SiC whisker diamond composite. This study describes the reactions of regenerating bone tissue to phagocytosable particles of diamond and SiC, using implanted bone harvest chambers in rabbits. The particles were dispersed in hyaluronan and introduced into a canal transversing the implant. The tissue that entered the canal during the following 3 weeks was then harvested. In previous studies using this model, particles of high density polyethylene, bone cement and chromium cobalt all caused an inflammatory reaction and a marked decrease in the amount of ingrown bone. In the present study, neither the diamond nor the SiC particles caused any decrease in bone formation. It appears that particles of diamond and SiC are comparatively harmless. PMID- 8730966 TI - Blood and cell compatibility of gelatin-carrageenan mixtures cross-linked by glutaraldehyde. AB - Mixtures of gelatin and iota-carrageenan cross-linked by glutaraldehyde were prepared and their physical properties and blood and cell compatibility were compared to gelatin as a control material. According to scanning electron microscopic observation of fracture surfaces, the mixtures were composed of dispersed and continuous domains which might be generated by phase separation of carrageenan. The thermal degradation temperature of iota-carrageenan in the mixtures rose with increasing gelatin content. The swelling process in the mixtures proceeded slower than in gelatin. Tensile strengths of the mixtures, except that containing 50% iota-carrageenan, increased with increased amounts of iota-carrageenan in the mixtures. The iota-carrageenan contents at the surfaces of the mixtures were generally higher than those admixed originally. Static friction coefficients of the mixtures were lower than that of gelatin. Plasma recalcification times of the mixtures were longer than that of gelatin. Platelet adhesion of the mixtures was lower than that of gelatin, while cell adhesion and growth assays using Chinese hamster ovary cells showed that cell adhesion and growth were not dependent on adding iota-carrageenan. It was concluded that blood compatibility of the mixtures increased and cell compatibility did not decrease, compared to gelatin. PMID- 8730967 TI - Biospecific interactions of Vitamin K-dependent factors with phospholipid-like polystyrene derivatives. Part I: Factor II. AB - Phosphorylated polystyrene derivatives with different compositions in phosphate groups were shown to be either recognized as phospholipidic or as DNA-like surfaces by antibodies from Systemic Lupus Erythematosus patients. In order to check whether these polymers were able to interact with Vitamin K-dependent coagulation factors, phosphorylated resins of various compositions in phosphate groups were assessed with regard to their interactions with Factor II, one of the Vitamin K-dependent factors. These studies were performed either in the presence or the absence of calcium ions, and with or without albumin precoating of the polymers. The results show that the affinity of the protein for the polymer is increased in the presence of calcium ions and depends on the composition of the polymer. The protein-polymer interactions involve the formation of binary or ternary complexes and the domains of predominance of these complexes were determined as a function of the calcium ion concentration in the assay. This allowed us to propose optimal conditions for Factor II purification by highly specific liquid chromatography using phosphorylated polystyrene resins of given compositions as stationary phases. PMID- 8730968 TI - In vitro biocompatibility of bioresorbable polymers: poly(L, DL-lactide) and poly(L-lactide-co-glycolide). AB - The acute toxicity of two degradable polymers, a 70:30 poly (L-D, L-lactide) (PLDLA) and a 90:10 poly(L-lactide-co-glycolide) (PLGA), was evaluated by the agar diffusion test and the filter test with L929 mouse fibroblasts. Extracts of the materials prepared in phosphate-buffered saline at 37 and 70 degrees C were assessed for mitochondrial succinate dehydrogenase activity by 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT assay) and the incorporation of 5-bromo-2'-deoxyuridine (BrdU) into DNA of BALB 3T3 cells. Both materials revealed no signs of cytotoxicity during the agar diffusions and filter tests. In the MTT and BrdU assays PLDLA and PLGA showed similar results. Cells treated with extracts prepared at 37 degrees C caused slight stimulation of mitochondrial activity. In contrast, cells incubated with the 70 degrees C media revealed a concentration-dependent decrease of mitochondrial activity. DNA synthesis was significantly decreased by the 37 degrees C extracts. As in the MTT assay, the effect of the extracts prepared at 70 degrees C was significantly greater. From these in vitro results it is suggested that PLDLA and PLGA have satisfactory biocompatibility. High concentrations of the degradation products, however, had a toxic influence on the cell culture systems used. PMID- 8730969 TI - Preparation and surface characterization of functional group-grafted and heparin immobilized polyurethanes by plasma glow discharge. AB - Functional group-grafted polyurethanes were prepared by oxygen plasma discharge treatment, followed by graft polymerization of 1-acryloylbenzotriazole (AB) and a subsequent substitution reaction of AB with sodium hydroxide and ethylene diamine. The primary amine or carboxylic acid groups grafted on the surfaces were coupled with heparin using water-soluble carbodiimide. The modified surfaces were characterized by measuring the water contact angle, electron spectroscopy for chemical analysis and attenuated total reflection Fourier-transform infrared spectroscopy. The amount of heparin covalently immobilized on the primary amine- and carboxylic acid group-grafted polyurethanes were 2.0 and 1.4 micrograms cm-2, respectively, as determined by the toluidine blue method. The water contact angle of the polyurethanes was decreased by AB grafting, and further decreased by the introduction of functional groups such as carboxylic acid and primary amine and immobilization of heparin, showing increased hydrophilicity of the modified surfaces. Heparin was almost not released from the immobilized surfaces in the physiological solution for 100 h, indicating good stability of immobilized heparin. PMID- 8730970 TI - Effects of cardiac output on disposition kinetics of sorbitol: recirculatory modelling. AB - 1. The purpose of this study was to determine the effects of cardiac output on distribution and elimination kinetics of the marker compound sorbitol. 2. The disposition kinetics of sorbitol were investigated after rapid intravenous injection and arterial sampling in nine patients who had undergone cardiac catheterization whereby the cardiac output was measured. 3. A minimal circulatory model consisting of pulmonary and systemic subsystems, both of which were characterized by an inverse Gaussian transit time density function, fitted the data very well. The method involves numerical inverse Laplace transform of the model equations. 4. The mixing clearance introduced as a novel non-compartmental parameter of distribution dynamics was significantly correlated with cardiac output. The steady-state volume of 14 l matched the extracellular volume. The systemic extraction ratio of 23% may reflect the fractional liver blood flow. 5. This pharmacokinetic model can be applied when an independent observation of cardiac output is available. In contrast to the conventional compartmental (or sum of exponential) approach it contains fewer adjustable parameters which can be more readily interpreted in physiological terms. PMID- 8730971 TI - Comparison of the effects of clonidine on tyramine- and methoxamine-evoked mydriasis in man. AB - 1. It has been reported previously that clonidine can potentiate tyramine-evoked mydriasis on the pain-free side of cluster headache patients. We examined whether a single oral dose of clonidine (200 micrograms) can also potentiate tyramine evoked mydriasis in healthy subjects, using mydriasis to methoxamine, a directly acting sympathomimetic amine, as a control. 2. Eight healthy male volunteers participated in four weekly sessions. In the first two sessions (Experiment 1) the effect of clonidine or placebo on the mydriasis to tyramine hydrochloride eyedrops (75 mM; 2 x 10 microliters), and in the last two sessions (Experiment 2) the effect of clonidine or placebo on the mydriasis to methoxamine hydrochloride eyedrops (20 mM; 2 x 10 microliters) was examined. In both experiments subjects were allocated to drugs and sessions according to a double-blind balanced design. In both experiments, pupil diameter of both the treated and the untreated eyes was recorded in standard ambient light and in the dark, before, and 2 h after clonidine/placebo, via binocular infrared television pupillometry. Salivation (dental roll technique), systolic and diastolic blood pressure (sitting), heart rate, and self-ratings of mood and feelings (visual analogue scales), were also measured before, and 2 h after the ingestion of clonidine or placebo. 3. Both tyramine and methoxamine produced a significant mydriasis, which was more prominent in the light condition (change in resting pupil size; mm +/- s.e.mean: tyramine/light 1.05 +/- 0.28; tyramine/dark: 0.73 +/- 0.15; methoxamine/light: 1.65 +/- 0.28; methoxamine/dark: 0.85 +/- 0.15). Clonidine produced a significant miosis in the untreated eye which was more prominent in the light condition (change in resting pupil size; mm +/- s.e.mean: Experiment 1, light: -1.34 +/- 0.19; Experiment 1, dark: -0.46 +/- 0.1; Experiment 2, light -0.97 +/- 0.18; Experiment 2, dark: -0.29 +/- 0.17). Clonidine had no significant effect on either tyramine- or methoxamine-evoked mydriasis. 4. In agreement with previous reports, clonidine significantly reduced salivation (g, mean +/- s.e.mean; Experiment 1: -0.84 +/- 0.22; Experiment 2: -0.55 +/- 0.11), systolic blood pressure (mm Hg; Experiment 1: -17.5 +/- 3.76; Experiment 2: -23.38 +/- 4.67), diastolic blood pressure (mm Hg; Experiment 2: -12.38 +/- 2.05), alertness (mm; Experiment 2: -24.19 +/- 5.40), and anxiety (mm; Experiment 1: -13.82 +/- 4.60), indicating the presence of pharmacodynamically effective tissue levels of the drug. 5. These results show that a single oral dose (200 micrograms) of clonidine causes significant miosis in human subjects, and fails to potentiate tyramine evoked mydriasis. This indicates that the pupil on the asymptomatic side of cluster headache patients is affected differently from the pupils of healthy volunteers by tyramine and/or clonidine. PMID- 8730972 TI - A comparison of ciprofloxacin, norfloxacin, ofloxacin, azithromycin and cefixime examined by observational cohort studies. AB - 1. The safety in everyday clinical usage of three 4-quinolone antibiotics, (ciprofloxacin, norfloxacin and ofloxacin), was compared with similar data for azithromycin and cefixime, each agent being examined by Prescription-Event Monitoring (PEM) during the early post-marketing period. 2. In PEM the exposure data are derived from general practitioner prescriptions confidentially provided by the Prescription Pricing Authority. Outcome data are provided by questionnaires (green forms) on which the prescribing medical practitioner records event data. When necessary, further information is obtained from a number of sources which include follow-up of all pregnancies and the patients' life-time medical record. 3. The main outcome measures were demographic information, including the patient's date of birth and sex; the indication for prescribing the drug being monitored; the reason for stopping treatment; the start and stop dates of treatment and the events recorded during and after treatment. 4. The final cohort for each of the five antibiotics exceeded 11000 patients. The only event significantly related to the use of all five antibiotics was nausea/vomiting. This was also the most frequent adverse event causing treatment to be discontinued with norfloxacin, ofloxacin and azithromycin (relevant information was not requested in the studies of ciprofloxacin and cefixime). Vaginal candidiasis was significantly more frequently associated with the use of the three 4-quinolones than with azithromycin and cefixime but it was frequently delayed until the week or two after the cessation of therapy. Within each event, as recorded in these studies, the highest event rates (the number of events per 1000 patients) in the week following the start of therapy were: 9.2 for diarrhoea with cefixime; 4.9 for nausea/vomiting with ofloxacin; 2.4 for rash with azithromycin; 2.2 for abdominal pain with norfloxacin; 1.5 for headache/migraine with ofloxacin; 1.4 for malaise/lassitude with ofloxacin; 1.2 for dizziness with norfloxacin. Uncommon events (reported in less than 1:1000 patients) included rare cases of allergic phenomena, convulsions and pseudomembranous colitis. There were no reports of tendinitis, tenosynovitis or tendon rupture in children but tendon disorders were reported in the two months following the start of treatment in 20 adults. A total of 307 pregnancies were reported. Thirty-eight of the 55 women who received these drugs during the first trimester of pregnancy gave birth to healthy babies. No congenital abnormalities were reported. Apart from one case of unconfirmed pseudomembranous colitis, none of the other 2468 deaths that occurred in these studies was attributed to the antibiotics. 5. These five antibiotics are acceptably safe antimicrobial agents when used in general medical practice. PEM is an effective method for monitoring the safety of recently introduced antimicrobial agents. PMID- 8730973 TI - Are young adults with asthma treated sufficiently with inhaled steroids? A population-based study of prescription data from 1991 and 1994. AB - 1. We conducted a descriptive cross-sectional study of asthma therapy among young adults to assess to what extent the current guidelines for asthma therapy have been implemented. In particular, we examined the use of inhaled corticosteroids in heavy users of inhaled beta-adrenoceptor agonists. 2. Data were retrieved from a population-based prescription database. For each of the years 1991 and 1994, all 20 to 44-year-olds who redeemed anti-asthma medication in the Odense area (210,000 inhabitants) were studied. 3. We identified the number of users and total sales volume for specific anti-asthma medications in defined daily doses (DDD) as well as the number of users and median annual doses of common regimens. Combined use of inhaled corticosteroids and inhaled beta-adrenoceptor agonists was also described. 4. The annual sales volume of anti-asthma drugs increased by 23% to 927,636 DDD from 1991 to 1994. Inhaled corticosteroids were mainly responsible for this with a 52% increase in number of users and an 88% increase in DDD. Inhaled beta-adrenoceptor agonists used in monotherapy remained the most popular regimen in 1994 (1685 users = 39%). Inhaled corticosteroids in combination with inhaled beta-adrenoceptor agonist were the second most popular regimen in 1994 (1308 users = 30%), increasing by 64% as compared with 1991. However, among patients with an annual use of inhaled beta-adrenoceptor agonist of 200 DDD (1600 "puffs') or more the percent of patients not receiving inhaled corticosteroids at all only fell from 37 to 33%. Though the number of patients being treated with inhaled corticosteroids has increased, there is still evidence of a substantial underuse. PMID- 8730974 TI - Pharmacokinetics of fluconazole in people with HIV infection: a population analysis. AB - 1. The population pharmacokinetics of fluconazole have been investigated in 113 male subjects with HIV infection and AIDS. Plasma concentration-time data (between 1 and 17 observations per dose) were collected from individuals as part of a pharmacokinetic investigation (13 subjects) or during routine fluconazole therapy (100 subjects) for the treatment or prophylaxis of fungal infection. 2. A one-compartment pharmacokinetic model was used to describe the disposition of fluconazole after oral and intravenous infusion doses. Population pharmacokinetic parameters were generated using the NONMEM and P-PHARM computer programs. 3. The population estimates (calculated using NONMEM) of fluconazole clearance and volume of distribution were 0.78 l h-1 and 47.61, respectively. The intersubject variability for these parameters was 41% and 8%, respectively. The model dependent estimate of the extent of absorption was 0.99 with an intersubject variability of 6%. Mean population estimates generated by NONMEM and P-PHARM were in close agreement. 4. Examination of the relationship between patient covariates and pharmacokinetic parameters indicated that intersubject variability in fluconazole clearance could in part be explained by the severity of disease (as indicated by CD4 + T-lymphocyte count) and renal function (indicated by estimated creatinine clearance). Other pharmacokinetic parameters were unaffected by these covariates. 5. Fluconazole clearance (estimated using NONMEM) in subjects with a CD4 + T-lymphocyte count less than and greater than 200 cells mm3 was 0.73 l h-1 (95% CI; 0.64-0.82 l h-1) and 0.99 l h-1 (95% CI; 0.86-1.12 l h-1), respectively. The regression model for fluconazole clearance that accounted for changes in renal function and disease severity was CL (l h-1) = 0.25 (33%) + 0.0057 (32%) x CLcr (in ml min-1) + 0.00068 (10%) x CD4 cell count (in cells mm-3) where intersubject variability (expressed as %CV) is shown in brackets. 6. Based on pharmacokinetic considerations a reduction in the dose of fluconazole would appear to be warranted in people with HIV infection who are seriously ill or who have compromised renal function. However, the emergence of resistance to fluconazole must also be considered when thinking of dosage adjustments. PMID- 8730975 TI - Pharmacodynamics of oxypurinol after administration of allopurinol to healthy subjects. AB - 1. Eight healthy subjects received 50, 100, 300, 600 and 900 mg allopurinol daily for 1 week each, in random order with 1 week separating each treatment period. The pre-dose plasma concentration of oxypurinol, the extent of inhibition of xanthine oxidase, plasma urate concentration and urine urate excretion rate were assessed on the last 2 days of each treatment week. 2. The ratio of 1-methyluric acid (1MU) over 1-methylxanthine (1MX) in the urine, following a dose of 50 mg 1MX infused intravenously over 20 min, was used to measure the inhibition of xanthine oxidase. 3. The steady-state plasma concentration of oxypurinol increased linearly with increasing dose of allopurinol between 50 mg to 600 mg day-1, with a weak indication of saturation at the higher 900 mg day-1 dose rate. 4. The relationships between plasma oxypurinol concentration and xanthine oxidase inhibition (1MU/1MX ratio), plasma urate concentration and urine urate excretion rate were fitted to an inhibition sigmoid Emax model and the C50 values for oxypurinol were 26.38 +/- 4.87, (mean +/- s.d.) 36.58 +/- 8.36 and 24.61 +/- 9.08 microM, respectively. 5. 1MU/1MX ratio appeared to be a reliable index of xanthine oxidase activity in vivo as the C50 for oxypurinol observed for 1MU/1MX ratio, plasma urate concentration and urine urate excretion rate were similar. 6. The concentration of oxypurinol required for inhibition of xanthine oxidase, as indicated by C50, was lower than those often observed in clinical practice. PMID- 8730976 TI - Stimulatory as well as inhibitory effects of ethinyloestradiol on the metabolic clearances of propranolol in young women. AB - 1. The metabolism of a single 80 mg oral dose of propranolol was determined in nine young women before and after administration of ethinyloestradiol alone (EE2) or in combination with norethindrone (OC). 2. Whereas the total clearance of propranolol (2713 +/- 404 ml min-1 (mean +/- s.e.mean)) was not significantly altered by either EE2 (3365 +/- 347 ml min-1) or the combined OC (2905 +/- 345 ml min-1), significant changes in all three primary metabolic pathways were observed. 3. The clearance through side-chain oxidation decreased from 345 +/- 55 ml min-1 to 262 +/- 33 ml min-1 after EE2 (P < 0.05). A similar reduction of cytochrome P450 metabolism by EE2 has been observed for other drugs. 4. The clearance through glucuronidation increased from 364 +/- 61 ml min-1 to 625 +/- 117 ml min-1 after EE2 (P < 0.01). Similar stimulation of glucuronic acid conjugation by EE2 has also been observed for other drugs. 5. The clearance through ring oxidation increased from 697 +/- 109 ml min-1 to 1280 +/- 162 ml min 1 after EE2 (P < 0.01). This observation appears to be a novel finding with EE2 and cytochrome P450 metabolism. 6. The treatment with OC produced changes in propranolol's metabolic clearances which were qualitatively similar to those generated by EE2. PMID- 8730977 TI - The influence of the sparteine/debrisoquine genetic polymorphism on the disposition of dexfenfluramine. AB - 1. To determine whether dexfenfluramine is a substrate of cytochrome P450 2D6 (CYP2D6), its disposition has been studied in nine extensive (EM) and eight poor metabolizers (PM) of debrisoquine. 2. Following a 30 mg dose of dexfenfluramine hydrochloride, urine was collected in all subjects for 96 h post-dose and plasma samples were collected in 11 subjects (six EMs and five PMs). Dexfenfluramine and nordexfenfluramine were measured in urine by h.p.l.c. and in plasma by g.c. 3. Urinary recovery of dexfenfluramine was greater in PMs than EMs (4136 +/- 1509 micrograms vs 1986 +/- 792 micrograms; 95% CI of difference 926-3374; P < 0.05) whereas that of nordexfenfluramine was similar in both phenotypes (PM: 1753 +/- 411 micrograms vs 1626 +/- 444 micrograms). 4. Dexfenfluramine AUC was higher in PMs (677 +/- 348 micrograms l-1 h) than EMs 359 +/- 250 micrograms l-1 h). The apparent oral clearance of dexfenfluramine was greater in EMs than PMs (93.6 +/- 42.4 l h-1 vs 45.6 +/- 19.5 l h-1; 95% CI of difference 1.2-94.7; P < 0.05). The renal clearance was similar in both phenotypes (EMs: 5.88 +/- 2.83 l h-1; PMs 6.60 +/- 2.01 l h-1), indicating that the higher urinary recovery of dexfenfluramine in PMs reflects higher plasma concentrations, rather than phenotype differences in the renal handling, of dexfenfluramine. 5. The apparent nonrenal clearance of dexfenfluramine was substantially lower (P < 0.05; 95% CI of difference 3.0-94.1) in PMs (39.0 +/- 19.5 l h-1) than EMs (87.6 +/- 41.2 l h 1). 6. There was a significant inverse correlation (rs = 0.776 95% CI-0.31-0.94; n = 11; p = 0.005) between the debrisoquine metabolic ratio and the apparent nonrenal clearance of dexfenfluramine. 7. PMs had a higher incidence of adverse effects (nausea and vomiting) than EMs. 8. In conclusion, the metabolism of dexfenfluramine is impaired in PMs. Thus CYP2D6, the isoenzyme deficient in poor metabolizers of debrisoquine, must catalyse at least one pathway of dexfenfluramine biotransformation. PMID- 8730978 TI - Fluconazole, but not terbinafine, enhances the effects of triazolam by inhibiting its metabolism. AB - 1. The interaction between triazolam and two antifungal agents, fluconazole and terbinafine, was investigated in a double-blind, randomized crossover study of three phases. 2. Twelve healthy young volunteers received 100 mg fluconazole, 250 mg terbinafine or placebo orally once a day for 4 days. On day 4 they took a single 0.25 mg dose of triazolam. Plasma samples were collected and pharmacodynamic effects were measured up to 17 h after the intake of triazolam. 3. Fluconazole increased the area under the triazolam concentration time-curve more than twofold (P < 0.001) and prolonged the elimination half-life of triazolam nearly twofold (P < 0.001). The peak concentration of triazolam was also increased significantly (P < 0.05) by fluconazole. 4. During the fluconazole phase pharmacodynamic effects of triazolam (e.g. digit symbol substitution test, body sway and drowsiness) were enhanced significantly (P < 0.05) when compared with the placebo phase. 5. Terbinafine did not change significantly the pharmacokinetics or pharmacodynamics of triazolam. 6. Care should be taken when triazolam is prescribed to patients using fluconazole. Although the interaction is not as strong as that of triazolam with ketoconazole or itraconazole, it is clinically significant. Triazolam and probably other drugs metabolized by CYP3A4 can be used in normal doses with terbinafine. PMID- 8730979 TI - Influence of concurrent antiepileptic medication on the pharmacokinetics of lamotrigine as add-on therapy in epileptic children. AB - 1. Lamotrigine is a new antiepileptic drug, chemically unrelated to currently used antiepileptic medication. Its pharmacokinetics can be influenced by concomitant antiepileptic medication. 2. This study was performed to assess the pharmacokinetic profile of lamotrigine in three groups of children treated with different types of comedication: drugs known to induce, to inhibit or to have no clinically significant influence on drug metabolism, respectively. 3. Thirty-one children aged 6 months to 5 years were included and received a 2 mg kg-1 single oral dose. Lamotrigine plasma profiles were different between the three comedication groups. The half-lives (mean +/- s.d.) were: 7.7 +/- 1.8 h, 21.9 +/- 6.8 h, 44.7 +/- 10.2 h in the "inducer', "other' and "inhibitor' groups respectively. 4. Patients were then dosed to steady state, with the dosage adjusted on the basis of the single dose pharmacokinetics to achieve a minimum plasma concentration between 1.5 and 3 mg l-1. The mean minimum plasma concentration for the three groups was 2.54 +/- 1.28 mg l-1 at steady state. 5. Dosage of lamotrigine can be optimised with knowledge of the metabolic effects of antiepileptic comedication. PMID- 8730980 TI - The effects of co-administration of benzhexol on the peripheral pharmacokinetics of oral levodopa in young volunteers. AB - 1. The effects of benzhexol on the absorption and pharmacokinetics of an oral dose of levodopa have been studied in 10 young healthy volunteers. Subjects were given a suspension of levodopa (250 mg) 90 min after either benzhexol (5 mg) or placebo in a randomized cross over design with doses separated by at least 1 week; on each occasion carbidopa was given 1 h before and 5 h after the dose of levodopa. Soluble paracetamol and radiolabelled DTPA were given with the levodopa as markers of gastric emptying. 2. Most subjects showed two peaks in the levodopa plasma concentration-time curve on the placebo day, with the second minor peak occurring 1-2 h after the dose. After benzhexol administration all subjects showed two or more peak levodopa concentrations in plasma. Benzhexol administration caused a significant decrease in the maximum concentration (43%; P < 0.05) of the initial peak and an increase (22%; P < 0.1) in the maximum concentration of the second peak. This change in absorption profile caused by benzhexol significantly altered the ratios of the second peak compared with the initial peak for both the maximum concentrations (P < 0.02) and for the AUC values (P < 0.05). Benzhexol administration did not affect the total AUC of levodopa (7.30 +/- 1.09 vs 7.19 +/- 1.26 micrograms ml-1 h; means +/- s.d.). 3. The plasma concentration-time curves for paracetamol showed similar profiles to those for levodopa and the ratios of the peak concentrations and AUC values for the second peak compared with the initial peak were increased significantly by benzhexol administration (P < 0.05). The total AUC of paracetamol was not affected by benzhexol administration (39.4 +/- 8.2 vs 40.0 +/- 8.9 micrograms ml 1 h; mean +/- s.d.) 4. Benzhexol altered the gastric emptying profile, shown by gamma-scintigraphy, with a reduced extent of initial emptying prior to the establishment of the plateau which is characteristic of levodopa administration in the fasting state. In consequence the ratio of the second to the initial phase of emptying was significantly higher (P < 0.01) following benzhexol treatment. 5. Benzhexol reduces the initial phase of gastric emptying after a dose of levodopa so that there is a decrease in the initial peak and a greater proportion of the dose is absorbed subsequently following the second phase of gastric emptying which occurs approximately 1 h later. Theoretically, this altered concentration time profile could be an advantage for some patients with Parkinson's disease. PMID- 8730981 TI - Inhibition of cytochrome P450 by nefazodone in vitro: studies of dextromethorphan O- and N-demethylation. AB - Nefazodone (NEF), a 5-HT2A/2C antagonist antidepressant, is extensively metabolized in the human body to hydroxy NEF (OH-NEF), p-hydroxy NEF (pOH-NEF), a dione metabolite, and via cleavage of the molecule to m-chlorophenyl-piperazine (mCPP) and BMY-33604. The latter is further metabolized to BMS-183695-01 (BMSa) and BMS-183562-01 (BMSb). To investigate the potential of NEF and its metabolites to interfere with the metabolism of other drugs, we tested these compounds for their ability to alter dextromethorphan (DMO) O-demethylation to dextrorphan (DOP; an index reaction for CYP2D6) and N-demethylation to 3-methoxy morphinan (MEM, a recently proposed index reaction of CYP3A3/4). The assay was performed in an in vitro system with human liver microsomes from three different donors. NEF, OH-NEF, pOH-NEF, mCPP and BMSb were weak inhibitors of DMO O and N-demethylation, with average Ki values ranging from 18 to 50 microM for DOP formation, and from 21 to > 200 microM for MEM formation. The dione metabolite and BMSa did not produce detectable inhibition of either pathway. The findings for DMO O demethylation, well-established as a CYP2D6-mediated reaction, indicate that NEF and metabolites are weak inhibitors of this reaction, with Ki values at least 100 times higher than fluoxetine (Ki = 0.1 microM +/- 0.09). The implications of results on DMO N-demethylation are not clear. In vivo data, as well as in vitro data based on "pure' CYP3A3/4 substrates, provide evidence for clinically relevant CYP3A3/4 inhibition by NEF, OH-NEF, and pOH-NEF. Thus, formation of MEM by N-demethylation of DMO may not constitute a suitable index reaction to probe CYP3A3/4 activity. PMID- 8730982 TI - Lack of effect of chloroquine on the debrisoquine (CYP2D6 and S-mephenytoin (CYP2C19) hydroxylation phenotypes. AB - The effects of chloroquine (CHQ) on debrisoquine hydroxylase (CYP2D6) and S mephenytoin hydroxylase (CYP2C19) were assessed in 11 black Zimbabwean and 12 white Swedish healthy volunteers. The activity of CYP2D6 was measured as the urinary debrisoquine to 4-hydroxydebrisoquine metabolic ratio and that of CYP2C19 as the urinary S- to R-mephenytoin enantiomer ratio (S/R). There were no statistically significant differences in either metabolic ratio as a result of prophylactic or loading doses of CHQ. This indicates that CHQ does not inhibit CYP2D6 or CYP2C19 in vivo and is unlikely to compromise the metabolism of substrates for these two enzymes. It is, therefore, also unlikely that residual CHQ in populations under study will interfere with phenotyping of either CYP2D6 or CYP2C19. PMID- 8730983 TI - Enantioselective disposition of hydroxychloroquine. PMID- 8730984 TI - Experimental pain techniques. PMID- 8730985 TI - The role of light scatter in the residual visual sensitivity of patients with complete cerebral hemispherectomy. AB - Various residual visual capacities have been reported for the phenomenally blind field of hemispherectomized patients, providing evidence for the relative roles of cortical and subcortical pathways in vision. We attempted to characterize these functions by examining the ability of five patients to detect, localize, and discriminate high-contrast flashed, flickering and moving targets. Dependent measures were verbal, manual, and oculomotor responses. As a control for light scatter, intensity thresholds for monocular detection of targets in the hemianopic field were compared with thresholds obtained when using an additional half eyepatch to occlude the blind hemiretina of the tested eye. One unilaterally destriate patient was tested on the same tasks. In photopic conditions, none of the hemispherectomized patients could respond to visual cues in their impaired fields, whereas the destriate patient could detect, discriminate, and point to targets, and appreciate the apparent motion of stimuli across his midline. Under reduced lighting, the threshold luminance required by hemispherectomized patients to detect stimuli presented monocularly was similar to that required for their detection when all visual information was occluded in the blind field, and only available to the visual system indirectly via light scatter. In contrast, the destriate patient's monocular threshold in his blind field was substantially lower than that for stimuli directly occluded in the blind field. As we found no range of stimuli which the hemispherectomized patients could detect or discriminate that was not also associated with discriminable scattered light, we conclude that the subcortical pathways which survive hemispherectomy cannot mediate voluntary behavioural responses to visual information in the hemianopic field. PMID- 8730986 TI - Spatial properties of retinal mosaics: an empirical evaluation of some existing measures. AB - Mosaics of neurons are usually quantified by methods based on nearest-neighbor distance (NND). The commonest indicator of regularity has been the ratio of the mean NND to the standard deviation, here termed the 'conformity ratio.' However, an accurate baseline value of this ratio for bounded random samples has never been determined; nor was its sampling distribution known, making it impossible to test its significance. Instead, significance was assessed from goodness-of-fit to a Rayleigh distribution, or from another ratio, that of the observed mean NND to an expected mean predicted by theory, termed the dispersion index. Neither approach allows for boundary effects that are common in experimental mosaics. Equally common are 'missing' neurons, whose effects on the statistics have not been studied. To address these deficiencies, random patterns and real neuronal mosaics were analyzed statistically. Ns independent random-point samples of size Np were generated for 13 Np values between 25 and 6400, where Ns x Np > or = 144,000. Samples were generated with rectangular boundaries of aspect ratio 1:1, 1:5, and 1:10 to examine the influence of sample geometry. NND distributions, conformity ratios, and dispersion indices were computed for the resulting 45,997 independent random patterns. From these, empirical sampling distributions and critical values were determined. NND distributions for small-to-medium, bounded, random populations were shown to differ significantly from Rayleigh distributions. Thus, goodness-of-fit tests are invalid for most experimental mosaics. Charts are presented from which the significance of conformity ratios or dispersion indices can be read directly. The conformity ratio reacts conservatively to extremes of sample geometry, and provides a useful and safe test. The dispersion index is nonconservative, making its use problematic. Tests based on the theoretical distribution of the dispersion index are unreliable for all but the largest samples. Random deletions were also made from 33 real retinal ganglion cell mosaics. The mean NND, conformity ratio, and dispersion index were determined for each original mosaic and 36 independent samples at each of nine sampling levels, retaining between 90% and 10% of the original population. An exclusion radius, based on a spatial autocorrelogram, was also calculated for each of these 10,725 mosaic samples. The mean NND was moderately insensitive to undersampling, rising smoothly. The exclusion radius was remarkably insensitive. The conformity ratio and dispersion index fell steeply, sometimes failing to reach significance while half of the cells still remained. For the same 33 original mosaics, linear regression showed the exclusion radius to be 62 +/- 3% of the mean NND. PMID- 8730987 TI - Limulus vision in the ocean day and night: effects of image size and contrast. AB - Male horseshoe crabs, Limulus polyphemus, use their eyes to locate mates day and night. We investigated their ability to detect targets of different size and contrast in a mating area of Buzzards Bay, Cape Cod, MA. We found that males can see large, high-contrast targets better than small, low-contrast ones. For targets of the same size, animals must be about 0.1 m closer to a low-contrast target to see it as well as a high-contrast one. For targets of the same contrast, animals must be approximately 0.2 m closer to a small target to see it as well as one twice as large. A decrease of 0.05 steradians in the size of the retinal image of a target can be compensated by a four-fold increase in contrast. About 60% of the animals detect black targets subtending 0.110 steradians (equivalent to an adult female viewed from about 0.56 m), while only 20% detect targets subtending 0.039 steradians. This study shows that horseshoe crabs maintain about constant contrast sensitivity under diurnal changes in light intensity in their natural environment. As a consequence of circadian and adaptive mechanisms in the retina, male horseshoe crabs can detect female-size objects about equally well day and night. PMID- 8730988 TI - Pigment granule migration in crustacean photoreceptors requires calcium. AB - We have investigated the role of calcium in the regulation of pigment granule migration in photoreceptors of the semi-terrestrial crab, Sesarma cinereum. Isolated crab eyes (eyecup plus eyestalk) were maintained in crustacean Ringer either prepared normally or calcium-free plus 50 mM EGTA. Pigment granule movement was indirectly observed by monitoring reflectance from the eye during light stimuli using intracellular optical physiological techniques. Electroretinograms (ERGs) were also measured during light stimuli. EGTA treatment caused gradual loss of centripetal migration of pigment granules (normally leading to pupillary closure), and photoreceptors eventually became locked in the open-pupil, dark-adapted state despite repeated stimuli. In contrast, ERG responses continued throughout EGTA treatment, although the size and shape of the response was altered. Normal ERG responses and pigment granule movements returned after replacing EGTA-Ringer with normal-calcium medium. These results suggest that centripetal migration of pigment granules in crustacean photoreceptors requires calcium. PMID- 8730989 TI - Basic visual capacities and shape discrimination after lesions of extrastriate area V4 in macaques. AB - Ibotenic acid lesions were made in four macaque monkeys in a region of cortical area V4 that corresponds to the lower quadrant of one hemifield. For visual testing, fixation locus was monitored with scleral search coils and controlled behaviorally to place test stimuli either in the lesioned quadrant or in a control location in the opposite hemifield. Some basic visual capacities were slightly altered by the lesions; there was a two-fold reduction of luminance contrast sensitivity as well as red-green chromatic contrast sensitivity, both tested with stationary gratings. On the other hand, little or no loss was found when contrast sensitivity for detection or direction discrimination was tested with 10-Hz drifting gratings nor was there a reliable change in visual acuity. Hue and luminance matching were tested with a spatially more complex matching-to sample task, but monkeys could not learn this task in the visual field locus of a V4 lesion. If previously trained at this locus, performance was not affected by the lesion. In contrast to the small effects on basic visual capabilities, performance on two form discrimination tasks was devastated by V4 lesions. The first involved discriminating the orientation of colinear groups of dots on a background of randomly placed dots. The second involved discriminating the orientation of a group of three line segments surrounded by differently oriented line segments. Some selectivity of the deficits for form discrimination was shown by the lack of an effect of the lesions on a global motion discrimination. These results show that while V4 lesions cause only slight disruptions of basic visual capacities, they profoundly disrupt form discriminations. PMID- 8730990 TI - Localization of AMPA-selective glutamate receptor subunits in the adult cat visual cortex. AB - We have studied the presence and distribution of alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionate (AMPA)-selective glutamate receptor subunits (GluR1, 2, 3, and 4) in the adult cat visual cortical areas 17, 18, 19, and the lateral suprasylvian areas (LSA). Reverse transcription-polymerase chain reaction (RT PCR) amplification indicated that the genes encoding GluR1, 2, 3, and 4 are expressed in these areas and Western blot analysis revealed that the size of the corresponding peptides is similar to those described in the rat brain. In situ hybridization (ISH) using digoxigenin-labeled riboprobes showed that mRNAs coding for GluR1 and GluR3 were located in cells in all layers of the areas examined and also in the underlying white matter. GluR1 mRNA was relatively abundant throughout layers II-VI while GluR3 mRNA revealed a more laminated pattern of expression, preferentially labeling cells in layers II, III, V, and VI. The distribution of AMPA-selective receptor subunit peptides was studied by immunohistochemistry using subunit specific antibodies and found to be consistent with ISH results. In addition, we observed that most of the cells strongly labeled by the anti-GluR1 antibody were non-pyramidal neurons and that intense GluR2/3 immunoreactivity was seen preferentially in pyramidal neurons. Interestingly, double-labeling experiments indicated that neurons expressing gamma-aminobutyric acid (GABA) as well as the GluR1 subunit were particularly abundant in deeper layers. The GluR4 peptide was predominantly found in a relatively low number of layer III and layer V neurons with either pyramidal or non-pyramidal morphology. Finally, the distribution of neurons expressing the various receptor subunits was similar in all the visual cortical areas studied. These findings indicate a high expression of GluR1-3 subunits in the cat visual cortex and that GluR1 and GluR2/3 subunits are particularly abundant in non pyramidal and pyramidal neurons, respectively. In addition, the results described here provide a reference for future studies dealing with the effect of visual deprivation on the expression of this receptor type. PMID- 8730991 TI - Characterization of protein phosphatases type 1 and type 2A in Limulus nervous tissue: their light regulation in the lateral eye and evidence of involvement in the photoresponse. AB - The activities of both protein phosphatases and protein kinases are responsible for the transient changes in the levels of phosphorylation and probably the functions of protein intermediates involved in the biochemical and physiological mechanisms underlying the photoresponse in photoreceptor cells from both vertebrate and invertebrate organisms. Of the known protein serine/threonine phosphatases, various forms of type 1 (PP 1) and type 2A (PP 2A) protein phosphatases are responsible for dephosphorylating many of the known phosphoproteins including those involved in photoreceptor cell function. In this report, we provide biochemical evidence for both PP 1- and PP 2A-like activities in the visual and nonvisual tissue of the horseshoe crab, Limulus polyphemus, that membrane and soluble forms of both enzymes are present, and that the activities of both enzymes are greater in light- than in dark-adapted lateral eyes. These activities were characterized using glycogen phosphorylase a, a substrate for both PP 1 and PP 2A, and various protein phosphatase inhibitors, including okadaic acid. We also report that okadaic acid, at concentrations required to inhibit PP 1, inhibited physiological functions of photoreceptor cells from the ventral eye, causing a delayed reduction of the resting membrane, and slowing and reducing light responses. PMID- 8730992 TI - A relationship between behavioral choice and the visual responses of neurons in macaque MT. AB - We have previously documented the exquisite motion sensitivity of neurons in extrastriate area MT by studying the relationship between their responses and the direction and strength of visual motion signals delivered to their receptive fields. These results suggested that MT neurons might provide the signals supporting behavioral choice in visual discrimination tasks. To approach this question from another direction, we have now studied the relationship between the discharge of MT neurons and behavioral choice, independently of the effects of visual stimulation. We found that trial-to-trial variability in neuronal signals was correlated with the choices the monkey made. Therefore, when a directionally selective neuron in area MT fires more vigorously, the monkey is more likely to make a decision in favor of the preferred direction of the cell. The magnitude of the relationship was modest, on average, but was highly significant across a sample of 299 cells from four monkeys. The relationship was present for all stimuli (including those without a net motion signal), and for all but the weakest responses. The relationship was reduced or eliminated when the demands of the task were changed so that the directional signal carried by the cell was less informative. The relationship was evident within 50 ms of response onset, and persisted throughout the stimulus presentation. On average, neurons that were more sensitive to weak motion signals had a stronger relationship to behavior than those that were less sensitive. These observations are consistent with the idea that neuronal signals in MT are used by the monkey to determine the direction of stimulus motion. The modest relationship between behavioral choice and the discharge of any one neuron, and the prevalence of the relationship across the population, make it likely that signals from many neurons are pooled to form the data on which behavioral choices are based. PMID- 8730993 TI - Glycine receptors in the rod pathway of the macaque monkey retina. AB - The distribution of glycinergic synapses in macaque monkey retina was investigated. The monoclonal antibody (mAb2b) against the alpha 1 subunit of the glycine receptor produced a punctate immunoreactivity that was localized to synapses. In central retina about 70% of the alpha 1 subunit-containing synapses were located in strata 1 and 2 of the inner plexiform layer, about 30% were located in strata 3 and 4, and immunoreactivity was absent in stratum 5. Electron microscopy showed that the majority of the synapses in strata 1 and 2 were on cone bipolar axons. The presynaptic profile always belonged to an amacrine cell. Presynaptic and postsynaptic profiles were further characterized using double label immunofluorescence with cell-type specific antibodies against calcium binding proteins. An antiserum against calretinin was used to label AII amacrine cells and an antiserum against recoverin was used to label flat midget bipolar cells. In the outer part of the IPL, 75% of the alpha 1-immunoreactive puncta were colocalized with calretinin-immunoreactive AII processes and 61% of the alpha 1-immunoreactive puncta were colocalized with recoverin-positive midget bipolar axons. These results suggest that the alpha 1 subunit of the glycine receptor is present at the chemical synapse made by AII amacrine cells with flat midget bipolar cells, thus providing a pathway for rod signals to reach midget ganglion cells. PMID- 8730994 TI - Glutamate immunoreactivity in the cat retina: a quantitative study. AB - Immunocytochemical methods were used to visualize glutamate immunoreactivity in the cat retina and to compare its localization with that of aspartate, GABA, and glycine. The cellular and subcellular distribution of glutamate was analyzed at the light-microscopic level by optical densitometry and at the electron microscopic level by immunogold quantification. The findings were consistent with the proposed role for glutamate as the neurotransmitter of photoreceptors and bipolar cells as particularly high concentrations of staining were found in synaptic terminals of these cells. Ganglion cells were also consistently stained. Aspartate was totally colocalized with glutamate in neuronal cell bodies but the synaptic levels of aspartate were much lower than for glutamate. In addition to the staining of photoreceptor, bipolar, and ganglion cells, glutamate immunoreactivity was also observed in approximately 60% of the amacrine cells. These cells exhibited colocalization with either GABA or glycine. The elevated levels of Glu in amacrine cells may reflect its role as a transmitter precursor in GABAergic cells and as an energy source for mitochondria in glycinergic cells. PMID- 8730995 TI - A glutamate-elicited chloride current with transporter-like properties in rod photoreceptors of the tiger salamander. AB - Glutamate, when puffed near the synaptic terminals, elicits a current in rod photoreceptors. The current is strongly dependent upon both the intracellular and extracellular chloride concentration: its reversal potential follows the predicted Nernst potential for a chloride permeable channel. The glutamate elicited current also requires the presence of extracellular sodium. This glutamate-elicited current is pharmacologically like a glutamate transporter: it is elicited, in order of efficacy, by L-glutamate, L-aspartate, L-cysteate, D aspartate, and D-glutamate, all shown to activate glutamate transport in other systems. Furthermore, it is reduced by the glutamate transport antagonists dihydrokainate (DHKA) and D,L-threo-3-hydroxyaspartate (THA). THA, when applied alone, elicits a current similar to that elicited by glutamate. The current cannot be activated by the glutamate receptor agonists kainate, quisqualate, NMDA and APB, nor can it be blocked by the glutamate receptor antagonists CNQX and APV. Thus, the current does not appear to be mediated by a conventional glutamate receptor. Taken together, the ionic dependence and pharmacology of this current suggest that it is generated by glutamate transporter coupled to a chloride channel. PMID- 8730996 TI - Functional architecture of area 17 in normal and monocularly deprived marmosets (Callithrix jacchus). AB - The organization of the primary visual cortex (VI) of the common marmoset (Callithrix jacchus) was studied both physiologically and by means of transneuronal labelling of geniculocortical afferents. We addressed the question whether monocular deprivation (MD) could stabilize segregation into ocular dominance (OD) columns, which are not seen in normal adult marmosets but are present in juvenile animals (Spatz, 1979, 1989). Properties of neurons in normal marmosets closely resembled those of other New-World and Old-World monkeys and orderly tangential progressions of preferred orientation were observed. However, in contrast to species that display well-defined OD columns, neurons of layer 4 in V1 of normal adult marmosets received balanced inputs from the two eyes. Early MD (even though followed by prolonged binocular experience into adulthood) resulted in a reduction of cell size in laminae of the lateral geniculate nucleus with input from the deprived eye and a dramatic overall shift in ocular dominance towards the nondeprived eye in the cortex. However, isolated clusters of cells dominated by the deprived eye were found in both layers 4 and 6. Injection of lectin-conjugated horseradish peroxidase (WGA-HRP) into the deprived eye revealed elongated patches of terminal label, about 350 microns wide, in flat-mounted sections through layer 4. Afferent segregation was sharper and more regular in the region of V1 representing parafoveal visual space than in that representing the fovea. Our findings support the notion that all Old-World and New-World monkeys possess the capacity for segregation of geniculocortical afferents into OD columns. PMID- 8730997 TI - Processing of color, form, and motion in macaque area V2. AB - We investigated the representation of color in cortical area V2 of macaque monkeys, and the association of color with other stimulus attributes. We measured the selectivity of individual V2 neurons for color, motion, and form. Most neurons in V2 were orientation selective, about half of them were selective for color, and a minority of cells (about 20%) were selective for size or direction. We correlated these physiological measurements with the anatomical location of the cells with respect to the cytochrome oxidase (CO) compartments of area V2. There was a tendency for color-selective cells to be found more frequently in the thin stripes, but color-selective cells also occurred frequently in thick stripes and inter-stripes. We found no difference in the degree of color selectivity between the different CO compartments. Furthermore, there was no negative correlation between color selectivity and selectivity for other stimulus attributes. We found many cells capable of encoding information along more than one stimulus dimension, regardless of their location with respect to the CO compartments. We suggest that area V2 plays an important role in integrating information about color, motion, and form. By this integration of stimulus attributes a cue invariant representation of the visual world might be achieved. PMID- 8730998 TI - Modelling the spatio-temporal modulation response of ganglion cells with difference-of-Gaussians receptive fields: relation to photoreceptor response kinetics. AB - Difference-of-Gaussians (DOG) models for the receptive fields of retinal ganglion cells accurately predict linear responses to both periodic stimuli (typically moving sinusoidal gratings) and aperiodic stimuli (typically circular fields presented as square-wave pulses). While the relation of spatial organization to retinal anatomy has received considerable attention, temporal characteristics have been only loosely connected to retinal physiology. Here we integrate realistic photoreceptor response waveforms into the DOG model to clarify how far a single set of physiological parameters predict temporal aspects of linear responses to both periodic and aperiodic stimuli. Traditional filter-cascade models provide a useful first-order approximation of the single-photon response in photoreceptors. The absolute time scale of these, plus a time for retinal transmission, here construed as a fixed delay, are obtained from flash/step data. Using these values, we find that the DOG model predicts the main features of both the amplitude and phase response of linear cat ganglion cells to sinusoidal flicker. Where the simplest model formulation fails, it serves to reveal additional mechanisms. Unforeseen facts are the attenuation of low temporal frequencies even in pure center-type responses, and the phase advance of the response relative to the stimulus at low frequencies. Neither can be explained by any experimentally documented cone response waveform, but both would be explained by signal differentiation, e.g. in the retinal transmission pathway, as demonstrated at least in turtle retina. PMID- 8730999 TI - Direction repulsion in motion transparency. AB - A series of experiments investigated perceived direction of motion and depth segregation in motion transparency displays consisting of two planes of dots moving in different directions. Direction and depth judgments were obtained from human observers viewing these "bi-directional" animation sequences with and without explicit stereoscopic depth information. We found that (1) misperception of motion direction ("direction repulsion") occurs when two spatially intermingled directions of motion are within 60 deg of each other; (2) direction repulsion is minimal at cardinal directions; (3) perception of two directions of motion always results in separate motion planes segregated in depth; and (4) stereoscopic depth information has no effect on the magnitude of direction repulsion, but it does disambiguate the depth relations between motion directions. These results are developed within the context of a two-stage model of motion transparency. On this model, motion directions are registered within units subject to inhibitory interactions that cause direction repulsion, with the outputs of these units pooled within units selective for direction and disparity. PMID- 8731000 TI - Relationship between numbers of retinal ganglion cells and lateral geniculate neurons in the rhesus monkey. AB - Studies of the numbers of retinal ganglion cells and lateral geniculate nucleus (LGN) neurons in primates suggest that the numbers of both types of neurons may vary over a two-fold range from one individual to another. This raises the question of whether the numbers of ganglion cells and LGN neurons are related or vary independently from individual to individual. We used stereological procedures to obtain unbiased estimates of the numbers of both cell types in seven rhesus monkeys. We found no significant correlation (rs = -0.21) between the numbers of retinal and LGN cells in the same animals. In agreement with previous studies, the average ratio of the number of retinal ganglion cells that project to the LGN and the number of LGN cells was approximately 1:1. However, this ratio varied over a two-fold range, from 0.78:1 to 1.64:1, in individual animals. These results have important implications for understanding the mechanisms of retino-geniculate development and for understanding the connectional wiring between the retina and LGN. PMID- 8731001 TI - A model for the effect of cigarette smoking on lung cancer incidence in Connecticut. AB - Population based data on smoking history derived from NCHS surveys were used to develop a model for lung cancer incidence in Connecticut. Trends in smoking prevalence suggest that, while the prevalence in men increased earlier than women, more male smokers have quit than their female counterparts. These trends in smoking prevalence suggest striking gender differences in a period effect for the smoking prevalence. Estimates of the proportion of current smokers, ex smokers, and the mean duration of smoking were used in a model for the lung cancer incidence rates. The form for the relationship between smoking history and the incidence rate for these subgroups was based on information from cohort studies. The models represented a mixture of the smoking subgroups where the effect of smoking was considered to be either a multiplicative effect on the underlying age distribution, or a separate effect in which the level of exposure was the sole contribution to risk among smokers. The multiplicative model explained more than 80 per cent of the deviance for the period and cohort effects, while the non-multiplicative model could only account for trends in females. Hence, these results suggest that a sizeable portion of the period and cohort contributions to the lung cancer incidence trends in Connecticut can be attributed to the multiplicative model that utilizes this smoking information, although the lack of more detailed information is a limiting factor in developing the model. PMID- 8731002 TI - Misclassification of smoking habits as a source of bias in the study of environmental tobacco smoke and lung cancer. AB - The relationship of environmental tobacco smoke to lung cancer risk in lifelong non-smokers is commonly studied using marriage to a smoker as the index of exposure. As smokers tend to marry smokers, relative risk estimates will be biased if some current or former smokers are misclassified as lifelong non smokers. This paper shows how various factors affect the magnitude of the bias and describes a method for obtaining misclassification-adjusted relative risk estimates. Application of the method to U.S. and Asian data for women suggests misclassification is an important determinant of the slight excess risk observed in non-smokers married to smokers. Reasons why our conclusions differ from those of others are discussed, as are other difficulties in interpreting the association between spousal smoking and lung cancer risk. PMID- 8731003 TI - Some techniques for the analysis of work sampling data. AB - Work sampling is an observational technique that produces counts representing the number of times that an individual has been observed performing each of several tasks. These data are collected using either systematic or random times of observation, and typically exhibit correlation between repeated observations on the same individual, with the degree of correlation being a function of the amount of time elapsed between measurements. Using several recently developed statistical techniques, we illustrate how it is possible to carry out analyses of these nominal outcomes that account for the correlation between repeated outcomes. We use description of a work sampling study to motivate the techniques and we compare empirically results from analyses based on several different underlying assumptions. PMID- 8731004 TI - A likelihood approach to meta-analysis with random effects. AB - In a meta-analysis of a set of clinical trials, a crucial but problematic component is providing an estimate and confidence interval for the overall treatment effect theta. Since in the presence of heterogeneity a fixed effect approach yields an artificially narrow confidence interval for theta, the random effects method of DerSimonian and Laird, which incorporates a moment estimator of the between-trial components of variance sigma B2, has been advocated. With the additional distributional assumptions of normality, a confidence interval for theta may be obtained. However, this method does not provide a confidence interval for sigma B2, nor a confidence interval for theta which takes account of the fact that sigma B2 has to be estimated from the data. We show how a likelihood based method can be used to overcome these problems, and use profile likelihoods to construct likelihood based confidence intervals. This approach yields an appropriately widened confidence interval compared with the standard random effects method. Examples of application to a published meta-analysis and a multicentre clinical trial are discussed. It is concluded that likelihood based methods are preferred to the standard method in undertaking random effects meta analysis when the value of sigma B2 has an important effect on the overall estimated treatment effect. PMID- 8731005 TI - The appropriateness of the Wilcoxon test in ordinal data. AB - In two-sample studies with ordinal responses, the Wilcoxon rank-sum test is generally chosen to test equality of the distributions, in spite of it being a specific test of location shift. I compared the power of the exact tests based on the Wilcoxon statistic, O'Brien's generalized Wilcoxon statistic, and the omnibus Smirnov statistic in the presence of location shift and scale alternatives. All three tests can gain power as a function of the scale parameter, depending on its magnitude relative to the shift parameter. As the relative influence of the scale parameter grows, the O'Brien test is the most powerful. I also compared the power of the asymptotic Wilcoxon test with its exact version, and found that it either inflates or deflates power if there are scale changes present. I present results of a simulation study. PMID- 8731006 TI - An asymptotic test for the equality of coefficients of variation from k populations. AB - The coefficient of variation is often used as a guide of the repeatability of measurements in clinical trials and other medical work. When possible, one makes repeated measurements on a set of individuals to calculate the relative variability of the test with the understanding that a reliable clinical test should give similar results when repeated on the same patient. There are times, however, when repeated measurements on the same patient are not possible. Under these circumstances, to combine results from different clinical trials or test sites, it is necessary to compare the coefficients of variation of several clinical trials. Using the work of Miller, we develop a general statistic for testing the hypothesis that the coefficients of variation are the same for k populations, with unequal sample sizes. This statistic is invariant under the choice of the order of the populations, and is asymptotically chi 2. We provide an example using data from Yang and HayGlass. We compare the size and the power of the test to that of Bennett, Doornbos and Dijkstra and a statistic based on Hedges and Olkin. PMID- 8731007 TI - The impact of patient compliance on drug concentration profile in multiple doses. AB - Physicians commonly prescribe drugs in a multiple dosage regimen for prolonged therapeutic activity. To study the effect of multiple dosing on drug concentration in blood, researchers often use deterministic models with the assumption that drugs are administered at a fixed dosage, with equal or unequal (fixed) dosing intervals. In practice, many patients do not comply with such a rigid schedule. Hence, two possible scenarios might occur: patients might not take the prescribed dosing amount, resulting in erratic dosing sizes; they might not adhere to the dosing schedule, resulting in erratic dosing times. We propose separate statistical models for these two scenarios and study their impact on blood serum/plasma concentration. With non-compliance, some basic concepts such as steady state need new definition. We provide a rigorous formulation for the principle of superposition which enables us to generalize the concept of steady state. Applying the proposed models, we demonstrate that non-compliance causes the drug concentration time curve to exhibit an increase in fluctuation. The increase in fluctuation due to non-compliance cannot be explained with use of the classical deterministic multiple dose model. PMID- 8731008 TI - Sample size for the exact conditional test under inverse sampling. AB - Inverse sampling is a sampling design in which one continues sampling subjects until one obtains a predetermined number of index subjects. This paper derives a procedure for calculation of the minimum required number of index subjects on the basis of the exact conditional test under inverse sampling. This paper studies quantitatively the effect on power calculations of the number of index subjects. To facilitate use of inverse sampling in study designs, this paper further provides a table that summarizes, in a variety of situations, the minimum required number of index subjects for powers equal to 0.90 and 0.80 at 0.05 level. It also includes a discussion on use of the approximation sample size formula derived on the basis of a variance-stabilizing transformation and large sample theory. PMID- 8731010 TI - Promotion of human sperm capacitation by superoxide anion. AB - Capacitation of spermatozoa is an essential procedure for fertilization. Capacitated spermatozoa have an increase in the intracellular cAMP and acrosome reaction (AR) occurs immediately. The effect of exogenous superoxide anion (O2-) on the level of intracellular cAMP and the percentages of both spontaneous AR and lysophosphatidylcholine-induced AR (LPC-AR) were studied using semen samples collected from 10 healthy and fertile volunteers working or studying in Lanzhou Medical College. Spermatozoa were separated by Percoll and incubated at 37 degrees C in Ham's F-10 medium with O2- generation system: xanthine+xanthine oxidase +catalase+diethylenetriaminepentaacetic acid+ sodium formate. The intracellular cAMP was determined by (3H)-cAMP radioimmunoassay at 3 h of incubation, and the percentages of AR and LPC-AR were evaluated by the triple stain technique at 3.5 h of incubation. The effects of SOD with different concentration were also determined. The results showed:the level of intracellular cAMP (pmol/10(8) spermatozoa) of spermatozoa increased from 14.0 +/- 1.3 to 23.2 +/- 2.5 (P < 0.01), and the percentages of AR and LPC-AR increased from 4.5 +/- 1.1% and 14 +/- 1.9% to 16 +/- 2.0% and 32.5 +/- 1.7%, respectively (P < 0.01 in both comparisons). SOD inhibited these processes concentration dependently. To investigate the source of O2- during in vivo sperm capacitation, female genital tract fluids collected from 6 healthy nonpregnant donors of reproductive age, and seminal plasma, capacitated and noncapacitated spermatozoa from 10 fertile volunteers were investigated by spin trapping method. The results showed: A typical electron paramagnetic resonance spectrum for O2- spin adduct was exhibited only in capacitated spermatozoa but not in vaginal or cervical secretions, uterine and fallopian tubal fluids, nor in seminal plasma and noncapacitated spermatozoa. These results suggested that only capacitated spermatozoa themselves are able to generate O2- which stimulated their capacitation in turn. Furthermore, on the basis of these data, we propose that it may be possible to utilize the inhibitory effect of SOD on sperm capacitation so as to regulate fertilization. PMID- 8731009 TI - The oxidative inactivation of sarcoplasmic reticulum Ca(2+)-ATPase by peroxynitrite. AB - The oxidative inactivation of rabbit skeletal muscle Ca(2+)-ATPase in sarcoplasmic reticulum (SR) vesicles by peroxynitrite (ONOO-) was investigated. The exposure of SR vesicles (10 mg/ml protein) to low peroxynitrite concentrations ( < or = 0.2 mM) resulted in a decrease of Ca(2+)-ATPase activity primarily through oxidation of sulfhydryl groups. Most of this deactivation (ca.70%) could be chemically reversed by subsequent reduction of the enzyme with either dithiothreitol (DTT) or sodium borohydride (NaBH4), indicating that free cysteine groups were oxidized to disulfides. The initial presence of 5 mM glutathione failed to protect the SR Ca(2+)-ATPase activity. However, as long as peroxynitrite concentrations were kept < or = 0.45 mM, the efficacy of DTT to reverse Ca(2+)-ATPase inactivation was enhanced for reaction mixtures which initially contained 5 mM glutathione. At least part of the disulfides were formed intermolecularly since gel electrophoresis revealed protein aggregation which could be reduced under reducing conditions. The application of higher peroxynitrite concentrations ( > or = 0.45 mM) resulted in Ca(2+)-ATPase inactivation which could not be restored by exposure of the modified protein to reducing agents. On the other hand, treatment of modified protein with NaBH4 recovered all SR protein thiols. This result indicates that possibly the oxidation of other amino acids contributes to enzyme inactivation, corroborated by amino acid analysis which revealed some additional targets for peroxynitrite or peroxynitrite-induced processes such as Met, Lys, Phe, Thr, Ser, Leu and Tyr. Tyr oxidation was confirmed by a significant lower sensitivity of oxidized SR proteins to the Lowry assay. However, neither bityrosine nor nitrotyrosine were formed in significant yields, as monitored by fluorescence spectroscopy and immunodetection, respectively. The Ca(2+)-ATPase of SR is involved in cellular Ca(2+)-homeostasis. Thus, peroxynitrite mediated oxidation of the Ca(2+)-ATPase might significantly contribute to the loss of Ca(2+)-homeostasis observed under biological conditions of oxidative stress. PMID- 8731011 TI - An EPR investigation of human methaemoglobin oxidation by hydrogen peroxide: methods to quantify all paramagnetic species observed in the reaction. AB - The method of Electron Paramagnetic Resonance (EPR) spectroscopy was used to study the reaction of human methaemoglobin (metHb) with hydrogen peroxide. The samples for EPR measurements were rapidly frozen in liquid nitrogen at different times after H2O2 was added at 3- and 10-fold molar excess to 100 microM metHb in 50 mM phosphate buffer, pH 7.4, 37 degrees C. Precautions were taken to remove all catalase from the haemoglobin preparation and no molecular oxygen evolution was detected during the reaction. On addition of H2O2 the EPR signals (-196 degrees C) of both high spin and low spin metHb rapidly decreased and free radicals were formed. The low temperature (-196 degrees C) EPR spectrum of the free radicals formed in the reaction has been deconvoluted into two individual EPR signals, one being an anisotropic signal (g parallel = 2.035 and g perpendicular = 2.0053), and the other an isotropic singlet (g = 2.0042, delta H = 20 G). The former signal was assigned to peroxyl radicals. As the kinetic behaviour of both peroxyl (ROO.) and non-peroxyl (P.) free radicals were similar, we concluded that ROO. radicals are not formed from P. radicals by addition of O2. The time courses for both radicals showed a steady state during the time required for H2O2 to decompose. Once all peroxide was consumed, the radical decayed with a first order rate constant of 1.42 x 10(-3) s-1 (1:3 molar ratio). The level of the steady state was higher and its duration shorter at lower initial concentration of H2O2. The formation of the rhombic Fe(III) non-haem centres with g = 4.35 was found. Their yield was proportional to the H2O2 concentration used and the centres were ascribed to haem degradation products. The reaction was also monitored by EPR spectroscopy at room temperature. The kinetics of the free radicals measured in the reaction mixture at room temperature was similar to that observed when the fast freezing method and EPR measurement at -196 degrees C were used. PMID- 8731012 TI - Inhibition of neutrophil-superoxide generation by alpha-tocopherol and coenzyme Q. AB - Effects of various derivatives of alpha-tocopherol (VE) and coenzyme Q (CoQ) on superoxide (O2.-) generation of neutrophils and protein kinase C (PKC) activity were examined. VE and CoQ8 inhibited O2.- generation of neutrophils stimulated by a protein kinase C mediated process monitored by cytochrome c reduction and spin trapping methods. The inhibitory action was observed not only with alpha tocopherol, but also with beta-, gamma-, delta-tocopherols and with tocol which is a chemical similar to VE but lacking methyl groups on the chromanol ring structure and which is not a radical scavenger. By contrast, no inhibition was observed with 2-carboxy-2, 5, 7, 8-tetramethyl-6-chromanol (CTMC, trolox) or 2, 2, 5, 7, 8,-pentamethyl-6-chromanol (PMC) which are water soluble VE derivatives having radical scavenging activity. Compounds having a similar isoprenoid chain, such as CoQ, also have inhibitory activity on PKC-dependent O2.- generation of neutrophils. The inhibitory activity of CoQ derivatives is dependent on the length of the unsaturated isoprenoid chain. CoQ derivatives having 16, 24 and 32 carbon isoprenoid chains corresponding to CoQ4, 6, and 8 inhibited O2.- generation but 4 and 40 carbon isoprenoid chains corresponding to CoQ2 and 10 had no inhibitory activity on O2.- generation. Alpha-tocopherol and CoQ inhibited PKC activity but the ID50 for O2.- generation and PKC activity was different for each compound. However, no direct relationship between VE content and O2.- generation of neutrophils was observed. These results suggest that isoprenoids of VE and CoQ participate in the inhibition of the NADPH oxidase activation system through modulation of the neutrophil membrane probably by the inhibition of PKC. PMID- 8731013 TI - Red cell vitamin E and oxidative damage: a dual role of reducing agents. AB - The purpose of this study was to determine the role of reducing agents in maintaining the integrity of vitamin E-deficient red cells. Three groups of one month-old male Wistar rats were fed a basal vitamin E-deficient diet supplemented with either 0, 10 or 100 mg d, 1-alpha-tocopheryl acetate per kg diet for up to 12 weeks. Washed red blood cells (5%) were resuspended in saline-phosphate buffer, pH 7.4, and were incubated at 37 degrees C with or without containing 12.5 mM 2, 2'-azobis (2amino- propane) dihydrochloride (AAPH), 2.8 mM glucose, 1 mM ascorbic acid, 10 mM hydrogen peroxide (H2O2), 250 microM dimethylsulfoxide (DMSO) or 2.8 mM deoxyribose (DR) for up to 20 hours. Addition of either glucose, AAPH, ascorbic acid or H2O2 markedly accelerated the rates of hemolysis and lipid peroxidation in the red cells of vitamin E-deficient rats. On the contrary, both glucose and ascorbic acid were protective against oxidative damage to the red cells of vitamin E-supplemented rats in a dose-dependent manner. Also, vitamin E supplemented red cells were more resistant to AAPH and H2O2 than the deficient cells. DMSO or. DR had no significant effects on the rates of hemolysis or lipid peroxidation. Glucose, but not others, maintained or slowed down the loss of glutathione (GSH) during incubation. The results obtained suggest a dual role of ascorbic acid and GSH in the function of vitamin E in maintaining red cell integrity: these reducing agents may exert antioxidant function by participating in vitamin E regeneration when certain levels of vitamin E is maintained, but promote oxidative damage by enhancing free radical generation when vitamin E is low or depleted. PMID- 8731014 TI - Mitochondrial biosynthesis controls the sensitivity of Chinese hamster cells to hydrogen peroxide. AB - The mechanism of H2O2-resistance of Hpr-4, a variant of Chinese hamster V79 cells, was investigated. The effect of H2O2 on the mitochondria of the parental and Hpr-4 cells was compared. First, both biochemical and ultrastructural results showed that mitochondria in the parental cells were damaged by exposure to H2O2, while those in Hpr-4 cells recovered from the damage. Second, the H2O2-resistance of Hpr-4 cells was reversibly reduced or recovered by the addition or removal of inhibitors of mitochondrial biosynthesis, respectively. Third, the parental cells were auxotrophic to pyruvate after exposure to H2O2. Fourth, H2O2-sensitivity of the parental cells was also enhanced by the inhibition of mitochondrial biosynthesis. From these results, it was concluded that the mitochondria of Hpr-4 cells apparently had a greater resistance to H2O2 than those of the parental cells and that functional mitochondria were involved in the recovery of Chinese hamster V79 cells from H2O2-induced damage. PMID- 8731015 TI - Catecholamines enhance dihydrolipoamide dehydrogenase inactivation by the copper Fenton system. Enzyme protection by copper chelators. AB - Catecholamines (CAs: epinephrine, norepinephrine, dopamine, L-DOPA, 6 hydroxydopamine) and o-diphenols (DOPAC and catechol) enhanced dihydrolipoamide dehydrogenase (LADH) inactivation by Cu(II)/H2O2 (Cu-Fenton system). The inhibition of LADH activity correlated with Cu(II), H2O2 and CA concentrations. Similar inhibitions were obtained with the assayed CAs and o-diphenols. CAs enhanced HO. radical production by Cu(II)/H2O2, as demonstrated by benzoate hydroxylation and deoxyribose oxidation; LADH counteracted the pro-oxidant effect of CAs by scavenging hydroxyl radicals. Captopril, dihydrolipoamide, dihydrolipoic acid, DL-dithiothreitol, GSSG, trypanothione and histidine effectively preserved LADH from oxidative damage, whereas N-acetylcysteine, N-(2 mercaptopropionylglycine) and lipoamide were less effective protectors. Catalase (though neither bovine serum albumin nor superoxide dismutase) protected LADH against the Cu(II)/H2O2/CAs systems. Denatured catalase protected less than the native enzyme, its action possibly depending on Cu-binding. LADH increased and Captopril inhibited epinephrine oxidation by Cu(II)/H2O2 and Cu(II). The summarized evidence supports the following steps for LADH inactivation: (1) reduction of LADH linked-Cu(II) to Cu(I) by CAs; (2) production of HO. from H2O2 by LADH-linked Cu(I) (Haber-Weiss reaction) and (3) oxidation of aminoacid residues at the enzyme active site by site-specifically generated HO. radicals. Hydrogen peroxide formation from CAs autoxidation may contribute to LADH inactivation. PMID- 8731016 TI - In vitro and in vivo induction of nitric oxide by murine macrophages stimulated with Bordetella pertussis. AB - Nitric oxide (NO) exhibits potent antimicrobial activity in vitro. The function of NO in host defenses in vivo, however, is presently unclear. Experiments were undertaken to determine the production of NO in vitro from murine peritoneal and alveolar macrophages, and murine macrophage cell line (J774A.1) stimulated with Bordetella pertussis or pertussis toxin (PT). In addition, we determined circulating levels of NO in the sera and bronchoalveolar lavage (BAL) fluids of mice infected intranasally with B. pertussis. The results of this study showed that in vitro murine peritoneal macrophages induce production of NO in response to B. pertussis and PT. In addition, murine macrophage cell line, J774A.1 also induces NO production after stimulation with B. pertussis. NO production was also detected in alveolar macrophages from mice infected intranasally with B. pertussis. Finally, a significant increment of circulating levels of NO was noted, in the sera but not in the BAL fluids, of mice infected intranasally with B. pertussis. PMID- 8731017 TI - Role of cell-associated N-acetyl-D-glucosamine specific haemagglutinin in the adhesion of Vibrio cholerae O1 to rabbit intestinal epithelial cells in vitro. AB - Previously a N-acetyl-D-glucosamine specific cell-associated haemagglutinin (HA) had been purified from a Vibrio cholerae O1 strain. This study documents the role of this purified HA as an adhesin of V. cholerae O1. A significant inhibition in the adhesion of V. cholerae O1 bacterial cells to isolated rabbit intestinal brush borders (RIBB) was observed when the latter were pretreated with purified HA in ELISA. Antibody raised against purified HA and Fab (IgG) fragment of this serum inhibited adhesion of the bacteria to isolated rabbit intestinal epithelial cells (RIEC). V. cholerae O1 (both Ogawa and Inaba serovars) showed less adherence to isolated RIEC of animals immunised with the purified HA. Patients convalescing from V. cholerae O1 infection showed high ELISA titres against the purified HA indicating that it is expressed in the host during the disease process. PMID- 8731018 TI - Stimulation of phagocytosis and phagosome-lysosome (P-L) fusion of human polymorphonuclear leukocytes by sulfatide (galactosylceramide-3-sulfate). AB - Recently, extensive attention has been paid to the physiological function of glycosphingolipids (GSLs) of mammalian cell membranes. Among a variety of GSLs, sulfatide (galactosylceramide-3-sulfate) has been proposed to be a specific receptor or binding molecule to microorganisms. However, no report has appeared on the direct stimulation by sulfatide for cellular function differentiation in phagocytic cells. We found that sulfatide showed a marked stimulation for phagocytic processes of human peripheral polymorphonuclear leukocytes (PMN) using heat-killed cells of Staphylococcus aureus coated with isolated lipid. Among mammalian acidic GSLs, sulfatide showed the highest stimulative activity for adhesion, phagocytosis and phagosome-lysosome (P-L) fusion by PMN. On the other hand, neutral GSLs did not stimulate essentially. Relative phagocytic rate of sulfatide-coated staphylococci was six times higher than that of the non-coated control and P-L fusion rate was ten times at maximum, respectively. Although the promotion mechanism of sulfatide for such phagocytosis or P-L fusion is not clear, it was strongly suggested that the existence of negative charges on carbohydrate moiety may be essential for the induction of differentiation of phagocytic cell function via signal transduction systems. PMID- 8731019 TI - Structural and immunochemical studies on the lipopolysaccharide of the 'T antigen'-containing mutant Proteus mirabilis R14/1959. AB - In DOC-PAGE, lipopolysaccharide (LPS) of Proteus mirabilis R14/1959 (Rb-type) mutant showed a ladder-like migration pattern indicating the presence of a high molecular weight polysaccharide chain. The isolated polysaccharide, called T antigen because of similarity with the T1 chain of Salmonella friedenau LPS, contained D-glucose, D-galacturonic acid (D-GalA), and D-GlcNAc in molar ratios 2:1:1 and was structurally different from the O-antigen of the parental S-strain P. mirabilis S1959 but identical to the O-antigen of another S-strain Proteus penneri 42. The importance of a D-GalA(L-Lys)-containing epitope, most likely present in the core region of LPS, and of GalA present in the T-antigen chain in manifesting the serological specificity of P. mirabilis R14/1959 were revealed using rabbit polyclonal homologous and heterologous R- and O-specific antisera and the appropriate antigens, including synthetic antigens which represent partial structures of various Proteus LPS. PMID- 8731020 TI - Contribution of interferon-gamma in protecting mice during pulmonary and disseminated infection with Cryptococcus neoformans. AB - In the present study, the role of interferon-gamma (IFN-gamma) in the host resistance against Cryptococcus neoformans was examined using a murine model of pulmonary and disseminated infection. In this model, mice were infected intratracheally with live yeast cells, and the histological changes in the lungs and the number of microorganisms in the lung and brain were compared in mice treated and untreated with anti-IFN-gamma monoclonal antibody (mAb) to define the contribution of endogenously synthesized IFN-gamma in the natural course of infection. Administration of this mAb reduced the accumulation of inflammatory cells in the alveolar septa, peribronchial and perivascular areas, and promoted the expansive growth of microorganisms in the alveoli and destruction of alveolar structure. The neutralization of endogenous IFN-gamma by mAb increased the number of microorganisms in the lung and brain, and significantly shortened the survival time of infected mice. On the other hand, administration of IFN-gamma decreased the number of microorganisms in these organs, and significantly extended their survival time. Considered together, our results suggest that endogenous IFN-gamma protects mice from infection with C. neoformans by inducing a cellular inflammatory response, potentiating the clearance of microorganism from the lungs and preventing its dissemination into the central nervous system. PMID- 8731021 TI - Purification and characterisation of a novel 34,000-Mr cell-associated proteinase from the dermatophyte Trichophyton rubrum. AB - A novel cell-associated proteinase was purified to homogeneity from cytoplasmic antigen preparations of Trichophyton rubrum by sequential isoelectric focusing and gel filtration chromatography. The enzyme exhibited relative molecular masses of 34,000-Mr (non-reduced sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE)), 15,000-Mr (reduced SDS-PAGE) and 37,000-Mr (substrate SDS-PAGE). It had a pH optimum of 7.5 and a pI of 4.5. The proteinase exhibited broad substrate specificity and it was strongly inhibited by the serine proteinase inhibitors phenylmethylsulfonyl fluoride and chymostatin. The N terminal amino acid sequence of the 34,000-Mr proteinase shared 50% homology with the deduced amino acid sequence of a Coccidioides immitis wall-associated chymotrypsin-type serine proteinase. This is the first cell-associated proteinase to be purified and characterised from T. rubrum and it would appear to be related to the chymotrypsin-type serine proteinases, a class of enzymes that have rarely been isolated from fungi. The function of the proteinase remains speculative although it may play a role in the development and subsequent proliferation of the fungus in vivo. PMID- 8731022 TI - Infectivity of preserved Cryptosporidium parvum oocysts for immunosuppressed adult mice. AB - The present study was undertaken to determine the infectivity of Cryptosporidium parvum oocysts for immunosuppressed adult C57BL/6N mice after the oocysts had been stored from 1-48 months at 4 degrees C in 2.5% potassium dichromate. All mice inoculated with oocysts 1-18 months old developed patent infections, while mice inoculated with older oocysts remained uninfected. The prepatent period was extended from 2 to 6 or 7 days as the storage time for oocysts increased. The finding that C. parvum oocysts remain infective for mice for at least 18 months offers important economic and time-saving advantages for investigators who frequently require large numbers of oocysts that must be painstakingly purified from calf manure. PMID- 8731023 TI - Karyotyping by PFGE of clinical isolates of Sporothrix schenckii. AB - From October 1991 to December 1992 we had eight patients with sporotrichosis at Tsukuba University Hospital in Japan. With 8 strains isolated from these patients, PFGE (pulsed-field gel electrophoresis) analyses were carried out to examine whether the karyotype of S. schenckii is distinguished by our method and whether this molecular approach is a useful means of biotyping of S. schenckii strains. Chromosomes were separated by contour-clamped homogeneous electric field (CHEF) gel electrophoresis. The strains had six to eight chromosomes and a total genome size was approx. 28 Mbp. Although these karyotypes of all the isolates looked closely similar to each other, they were grouped into three types. PMID- 8731024 TI - Purification of a product from Salmonella typhimurium with the ability to inhibit mitogen-induced proliferation of murine splenic T-lymphocytes. AB - We attempted to purify a substance that inhibits mitogen-induced proliferation of murine splenic T-lymphocytes from Salmonella typhimurium. The soluble fraction of a suspension of bacteria disrupted by sonication was chromatographed serially on Mono Q HR, Superdex 200 HR and HiLoad Superdex 75 p.g. columns. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis revealed that the purified active substance migrated as a single band corresponding to a molecular mass of 87 kDa. We designated the purified substance S. typhimurium-derived inhibitor of T-cell proliferation (STI), which, at 0.2 microgram/ml and above, inhibited proliferation and augmented CD25 expression of phytohemagglutinin-stimulated murine splenic lymphocytes. These findings suggested that the immunosuppression induced by Salmonella infection may be attributable to STI. PMID- 8731025 TI - Low affinity, antibody binding of an Escherichia coli-derived component. AB - This investigation describes the detection of a component in Escherichia coli capable of binding a large proportion of human antibody variable domains including otherwise highly monospecific antibodies induced by an in vivo antibody response. This interaction is of low affinity, but cross-linking of IgG molecules by, e.g. anti-immunoglobulin preparations, provides a sufficient degree of multivalency to promote a high avidity interaction. This binding which occurs both with kappa and lambda light chain-containing antibodies, appears to involve the variable region of human antibodies making it a superantigen-like activity. This is proposed based on the facts that: (i) different human antibodies of IgG1 isotype appear to bind to different extents suggesting that variable domain differences determine the binding activity; and (ii) addition of soluble antigen abrogates the interaction with the E. coli-derived molecule. Future studies of the nature and possible in vivo consequences of these interactions are warranted since any superantigen activity associated with this binding might affect the human immune response occurring as a consequence of E. coli infections. PMID- 8731026 TI - Epitope mapping the Fim2 and Fim3 proteins of Bordetella pertussis with sera from patients infected with or vaccinated against whooping cough. AB - Antibody-binding epitopes on the Fim2 and Fim3 proteins of Bordetella pertussis, which have been associated with the induction of protective antibody, were located using sera from 12 patients with whooping cough and 4 vaccinated children. Fifteen epitopes were identified on both Fim2 and Fim3. In each case 9 were recognised by serum antibody from 11 or more infected patients. Epitopes associated with the highest IgG activity were not the same as those associated with the highest IgA activity. None of the vaccinated patients had detectable IgA. Most epitopes showed little or no evidence of serotype-specific responses, suggesting this is largely directed towards conformational epitopes. The reactivity of all but two epitopes was confirmed in an ELISA with patients' sera in which epitopes were re-synthesised as free soluble peptides. The short linear epitopes described may therefore be useful in the development of serodiagnostic assays but are unlikely vaccine candidates. PMID- 8731028 TI - Glutaraldehyde permeation: choosing the proper glove. AB - Six different gloves were tested with five different aqueous glutaraldehyde formulations to determine each glove's resistance to permeation. When tested against 2% or 3.4% glutaraldehyde solutions, nitrile rubber, butyl rubber, a synthetic surgical glove, and polyethylene were each impermeable for at least 4 hours. The two latex gloves tested showed glutaraldehyde breakthrough at 45 minutes. When the latex gloves were doubled, the time to first breakthrough increased to 3 to 4 hours. With 50% glutaraldehyde, only butyl rubber and nitrile rubber were impermeable for extended periods. The surgical synthetic glove had breakthrough at 1 hour, whereas polyethylene and the two latex gloves had breakthrough in less than 1 hour. PMID- 8731027 TI - Should infection control practitioners do follow-up of staff exposures to patient blood and body fluids? AB - BACKGROUND: The purpose of this study was to determine the efficiency of a joint infection control/occupational health program for the follow-up of accidental blood or bloody body fluid exposures in health care workers. METHODS: A comprehensive staff follow-up program for all blood exposures with known patient sources was initiated in 1989, consisting of patient follow-up by the Infection Control Department (risk assessment for hepatitis B virus [HBV] and [HIV] infection and obtaining of consent for HIV testing) and staff follow-up by the Occupational Health Department. In 1992 a mailed survey was conducted to examine exposure follow-up policies and responsibilities in large teaching hospitals across Canada. RESULTS: A total of 924 blood exposures with known patient sources were reported between January 1989 and December 1993. HIV and HBV screening was obtained for 67.9% and 87.6% of patients assessed as at low risk and 82.3% and 92.2% of those assessed as at high risk for infection, respectively. Two previously unknown HIV-seropositive patients were identified, one of whom had been classified as at low risk (one of 530 [0.19%] patients at low risk who underwent screening). Primary reasons for screening being missed were patient discharge (46.3%) or communication problems (18.0%). The requirement for informed written consent before HIV screening accounted for the difference in completed HIV and HBV screens. Results of the hospital survey indicated that 40.8% of Canadian hospitals follow up all patients who are involved in blood exposures; however, most hospitals still rely on the physician to obtain consent (87.6%). CONCLUSIONS: Use of ICPs to screen patients involved in staff blood exposures during regular hours may be the most efficient method of follow-up, particularly if supplemented by a backup team of health professionals on nights and weekends. Although screening all patients for HBV/HIV may detect patients with undisclosed high-risk behaviors, institutions must decide whether the practice is cost effective in areas of low prevalence. PMID- 8731029 TI - Tuberculosis control in health care workers: an algorithmic approach. AB - Health care workers have always been at risk for contracting tuberculosis (TB) from patients with active disease. In addition, health care workers who have active TB pose a risk for transmitting TB to patients. As a result, institutions that employ health care professionals must adopt programs to reduce the probability of transmission of TB to their employees or their patients. This article discusses the new guidelines for preventing TB issued by the Centers for Disease Control and Prevention and suggests approaches for adapting the guidelines to the needs of individual institutions. It emphasizes the importance of skin testing for early detection, correct interpretation of the tests, the approaches to determining who should be tested, and the relative frequency with which employees should be tested. It presents algorithms to assist employee health and infection control personnel in screening current and prospective employees, and in responding to positive and negative test results, and the booster effect. PMID- 8731031 TI - Early ovarian cancer: a review of its genetic and biologic factors, detection, and treatment. AB - Early-stage ovarian carcinoma requires comprehensive surgical staging; reexploration for patients who had suboptimal initial surgery would indicate an apparent early ovarian carcinoma. After proper surgery, patients can be subdivided into a high- or low-risk group, and treatment options then can be discussed with the patient. Patients in the low-risk category can be followed up expectantly without any form of adjuvant therapy. Patients in the high-risk category, however, should be encouraged to participate in randomized clinical trials, because it is unclear at the current time which combination of chemotherapy and how many treatments should be used. A platinum-based paclitaxel regimen probably should be used, although the relative merits of carboplatin and cisplatin and the appropriate schedule for taxol (1 hour vs. 3 hours vs. 24 hours vs. 96 hours) are yet unknown. It is hoped that clinicians will continue to encourage patients to participate in randomized clinical trials so that optimal therapy for early ovarian cancer can be established. PMID- 8731033 TI - Three-dimensional ultrastructure of the surface of the tongue of the rat snake, Elaphe climacophora. AB - BACKGROUND: Many studies have been performed to clarify the relationship between behavioral performance of the tongue and Jacobson's organ. The purpose of the present study was to examine the ultrastructural features of the surface of the tongue of the rat snake, Elaphe climacophora, and to delineate the functional relationship between the tongue and Jacobson's organ from a morphological perspective. METHODS: The three-dimensional ultrastructure of the surface of the tongue of the rat snake Elaphe climacophora was investigated by scanning electron microscopy. RESULTS: Most of the surface of the bifurcated apex of the tongue was relatively smooth. Dome-shaped, hemispherical bulges or microfacets were compactly arranged on the epithelial cell surface over this entire region. Intercellular borders were clearly recognizable as striations. These features were almost the same as those of the dorsal surface of the transitional area between the bifurcated lingual apex and the anterior part of the lingual body. In the posterior half of the lingual body, no microfacets were seen at all. Both microridges and microvilli were compactly distributed on cell surfaces. CONCLUSIONS: No evidence was obtained from our ultrastructural analysis for an important role of the lingual apex in the vomeronasal system. By contrast, the epithelial surface of the body of the tongue appeared suitable for retaining stimulating compounds. PMID- 8731032 TI - Sequestration of mitotic (M-phase) chromosomes in autophagosomes: mitotic programmed cell death in human Chang liver cells induced by an OH* burst from vanadyl(4). AB - BACKGROUND: Fragmentation of genomic DNA in apoptosis/programmed cell death (PCD) is a characteristic hallmark in which both 2N and 4N DNA from G1, S, and G2/M cell cycle phases were seen degraded to the sub-2N Ao level in PCD such as from serum deprivation, glucocorticoid treatment, and gamma-radiation. However M-phase (mitotic) cells are said to perish only via non-programmed or necrotic cell death unless they were allowed to complete cytokinesis and re-enter interphase. The morphological criteria of PCD refer only to interphase cells with intact nuclear membranes, none seems applied to mitotic cells. We show here autophagic sequestration of mitotic chromosomes in a typical PCD response where G1, S, and G2/M DNA were replaced by a sub-2N Ao peak, suggesting that mitotic cells may yet have the option of PCD or suicide. Autophagy is absent in necrosis. METHODS: Mitotic human Chang liver cells in normal monolayer culture were compared with apoptotic counterparts initiated by a burst of hydroxyl free radicals (OH*) generated from vanadium internalized by an NH4Cl prepulse containing vanadyl(4) ions. Total (free and bound) vanadium uptake was quantitated by elemental spectral analysis of single cells using a) Particle-Induced X-ray Emission (PIXE) profiling, and b) Scanning Transmission Ion Microscopy (STIM) in the nuclear microscope. The Coulter EPICS PROFILE II flow cytometer was used for a) the cell cycle analysis using propidium iodide-DNA binding, b) intracellular pH (pHi) evaluation in the acidification-and-recovery cycle, using ratiometric 2',7'-bis(2 carboxyethyl)-5 (and-6)-carboxyfluorescein (BCECF) fluorescence quantitation. Transmission electron microscopy examined the morphological changes. Vanadyl(4)- generated hydroxyl free radicals (OH*) were evaluated by measuring OH*-benzoic adduct fluorescence at 304/413 nm using the SPEX Fluoromax photon counting spectrofluorometer. RESULTS: Nuclear microscopy showed that a 30 min acidification prepulse containing 4mM vanadyl(4) ions, V(4), had increased the total (free and bound) vanadium concentration of human Chang liver cells from normal ultratrace levels to 56,992 ppm of dry wt (1.1174 Eq per kg dry wt). After the prepulse, cells realkalinized in DMEM growth medium, recovering to the physiological pHi level in 30 min. At the physiological pH 7 level, V(4) generated a burst of OH* free radicals in the order of 15,000 folds above the prepulse (pH 4.5) level. In these conditions, spectrofluorometric evaluation showed loss of DNA intercalation with propidium iodide (PI-DNA binding) indicating DNA degradation. Cell-by-cell evaluation of the PI-DNA binding by flow cytometry showed abolition of G1, S, and G2/M phases and their replacement by a sub-2N Ao peak of fragmented DNA, emulating serum deprivation PCD in these cells. Immediately upon initiating an OH* burst ultrastructural profiles showed mitotic chromosomes (M-phase chromatin) being surrounded by rough endoplasmic reticulum (RER) and small vesicles, indicating their sequestration in autophagosomes. Autophagy was also seen to be a prominent feature in serum deprivation PCD. CONCLUSION: Sequestration of mitotic chromosomes by autophagosomes in a typical PCD response showed a well-defined morphological pathway for direct degradation of M-phase chromatin without first completing cytokinesis. Mitotic cells could commit suicide via autophagy directed at its own chromatin. Autophagic sequestration of chromatin in PCD is novel. PMID- 8731034 TI - Immunocytochemical localization of melatonin in the harderian gland of Syrian hamster. AB - BACKGROUND: The Harderian gland (HG) is a tubulo-alveolar gland found within the ocular orbit of animals which present a nictitating membrane. The Harderian gland is regarded as an extrapineal melatonin producing organ and both, photoperiod and melatonin have been shown to exert an important role in the metabolism and morphological features of such a gland. Our results seem to support the presence of melatonin in the nuclei of the HG cells, although our studies have not definitively proved such presence. METHODS: An Immunocytochemical anti-melatonin technique was done over free sections of Bouin fixed material obtained from Syrian hamsters. Some of the sections were embedded in an epoxy resin and studied under electron microscope. RESULTS: The presence of positive immunoreaction was observed at the level of the nuclear membranes and in close relation to chromatin. No differences were observed between males and females nor between pinealectomized animals and control ones. CONCLUSIONS: Our results suggest the binding of melatonin to the cell nucleus in all cell types of the gland. These observations are in accord with the binding studies performed by Acuna Castroviejo in purified cell nuclei of rat liver (Acuna-Castroviejo et al., 1994. J. Pineal Res., 16:100-112) and the earlier one by Menendez-Pelaez et al. (1993a,b, J. Pineal Res., 15:59-69; J. Cell Biochem., 53:373-3*2) using the light microscope. Our results seem to support the idea of a nuclear action of melatonin and they agree with the observations of Carlberg and Wiesenberg (1995, J. Pineal Res., 18:171-178) about the activation of some orphan receptors by melatonin. PMID- 8731035 TI - Tissue distribution and structure of barrier cells in the hematopoietic and lymphoid organs of salmonids. AB - BACKGROUND: Barrier cells have been recognized as a discrete group of fibroblastic- or myofibroblastic-like cells located in the lymphoid and hematopoietic organs of mammals. This paper reports the results of a morphological study of the main lymphoid organs of three salmonid species, in which cells structurally similar to the mammalian barrier cells were observed in healthy animals. METHODS: The spleen, kidney, and thymus of fingerlings of rainbow trout, Oncorhynchus mykiss, and Coho salmon, Oncorhynchus kisutch, and of adult brown trout, Salmo trutta fario, were processed for electron microscopy study using various fixation methods. Semithin sections were used for the Periodic Acid-Schiff (PAS) staining technique, and for the demonstration of the endogenous peroxidase activity. RESULTS: The kidney and spleen of all the species contained a variable, but usually low, number of electron-dense, elongated, and branched cells, ultrastructurally similar to the mammalian barrier cells. They also occurred in the thymus of some brown trout and Coho salmon, but not of rainbow trout. The electron density of this cell type was present after the various types of fixation procedures. They show numerous ribosomes, well developed secretory organelles, electron-clear vesicles, large granules, and microfilaments. In all the salmonid species, barriers cells were positive for PAS staining and for endogenous peroxidase activity. A small number of barrier cells were in mitosis. In the different organs barrier cells appeared as isolated cells, or forming syncytial networks. They were found lining the blood sinusoids of the splenic red pulp and of the renal hematopoietic tissue, in the periellipsoidal sheaths, and closely associated with erythropoietic and plasmacytopoietic foci. CONCLUSIONS: Our results demonstrate the presence of cells closely resembling mammalian barrier cells in the hematopoietic and lymphoid organs of salmonids. The structure and tissue distribution of the salmonid barrier cells are discussed in relation to the functional roles described for this cell type in mammals. PMID- 8731037 TI - Streaming conjunctiva. AB - BACKGROUND: Epithelial proliferation is linked with cell displacement. When a cell divides, one of its progeny replaces the dividing ancestor and the other is displaced into a more remote location that has to be vacated first by peripheral cells. As cells are neither pushed nor pulled in a mechanical sense, and since they do not move by their own means, this displacement is best regarded as streaming. The purpose of the present study was to measure epithelial cell streaming in adult rat conjunctiva. METHODS: Twenty-seven female adult rats were injected i.p. with 18.5 KBq [3H]-thymidine/g body weight, specific activity 185 GBq/mMol. Three rats were killed at different times up to 28 d. Eyes and eyelids were removed in one piece, cut along the pupillary-optic nerve line into 5 microns-thick sections, and prepared for autoradiography. In each eye, the entire upper conjunctiva extending from limbus to the palpebral muco-cutaneous junction was scanned with an ocular micrometer grid. The limbus served as point of origin. The x, y coordinate of each nucleus with two grains or more and its grain content were recorded. RESULTS: One hour after labeling, labeled cells were spread evenly along the basal layer. Cells of the upper layers were not labeled. As time passed by, labeled cells in the fornix became more abundant, while in the limbus and palpebral margin, their frequency declined. Labeled cells streamed from the limbus and palpebral muco-cutaneous junction to the fornix. Bulbar conjunctival epithelia streamed at a velocity of 13.2 microns/day. Epithelia in the palpebral conjunctiva streamed at a velocity of 11.8 microns/day. At the same time cells streamed from the basal layer to the epithelial surface at a velocity of 0.4 microns/day. Generation time was 3.9 days. CONCLUSIONS: Bulbar and palpebral conjunctivae are two independent cell kinetic systems that originate in two stem cell regions, one in the limbus and the other in the muco-cutaneous junction. Each system is made of two compartments, a progenitor where cells proliferate, and a compartment of non proliferating end cells. Progenitors created in the first, enter the second and turn into end cells. Ultimately they die in the fornix. The undetermined limbus stem cell generates two epithelial cell lines, a corneal and a conjunctival. PMID- 8731038 TI - Peripheral blood mononuclear cells develop into multinucleated osteoclasts in tissue culture. AB - BACKGROUND: Previous studies have shown that osteoclasts are derived from mononuclear cells of hemopoietic bone marrow and peripheral blood. The purpose of this study was to demonstrate the presence of multinucleated osteoclasts after adding mononuclear cells from peripheral blood into established explants of fetal mouse calvaria in vitro. METHODS: In order to utilize osteoclast-free bone, the fetal calvariae were obtained from 13-14-day pregnant Swiss Webster mice and cultured in BGJb medium for 9 days. At day 9, peripheral blood mononuclear cells were isolated as a light density layer from adult Swiss Webster mice with the Ficoll-Paque density gradient separation method and co-cultured with the osteoclast-free, fetal mouse calvaria. RESULTS: After 10 days of co-culture, multinucleated cells, which have all the characteristics of osteoclasts, were found in juxtaposition to seams of woven bone. Two multinucleated osteoclasts per one million light density peripheral blood mononuclear cells were found in the experimental group; none were found in the mononuclear cell-free control group. CONCLUSIONS: Peripheral blood mononuclear cells can give rise to multinucleated osteoclasts in developing bone in vitro but will not develop without bone. PMID- 8731036 TI - Mouse Meckel's cartilage chondrocytes evoke bone-like matrix and further transform into osteocyte-like cells in culture. AB - BACKGROUND: We reported that when Meckel's cartilage was transplanted ectopically, chondrocytes transformed into osteocyte-like cells accompanying the extracellular calcified matrix. However, we could not determine whether the osteocyte-like cells were derived from host tissues or from Meckel's cartilage itself. Therefore, we examined whether the Meckel's cartilage chondrocytes, which have a retrogressive ultimate fate, are capable of inducing the observed calcification and further transform into osteocyte-like cells in culture. METHODS: Meckelian chondrocytes isolated enzymatically were plated at a low density and grown in alpha-MEM containing 10% FBS at 37 degrees C under 5% CO2 in air for up to 4 weeks. RESULTS: Chondrocytes were fibroblast-like cells early in culture, but gradually transformed from polygonal cells into typical chondrocytes showing metachromasia with toluidine blue staining. After an additional week of culture, the chondrocytes transformed from large to small round cells accompanying nodule formations. Small round cells multiple-layered actively, and showed more intense alkaline phosphatase (ALPase) activity. Immunostaining identified type II collagen in the extracellular matrix at 2 weeks of culture, and type I collagen and osteocalcin were later synthesized by round cells. von Kossa's reaction showed extensive precipitation of calcification throughout the flocculent materials. Ultrastructural analysis showed that the cells surrounded by calcified matrix strongly resembled osteocytes. CONCLUSIONS: The present study suggested that the Meckel's cartilage chondrocytes can express the osteocyte-like phenotype in vitro during synthesis of bone-type marker proteins such as osteocalcin or type I collagen. PMID- 8731039 TI - Basic fibroblast growth factor within endothelial cells during vascularization of the anterior pituitary. AB - BACKGROUND: Basic fibroblast growth factor (FGF-2), a potent angiogenic peptide, is known to be present in gonadotropes of the anterior pituitary parenchyma of rats and mice, and has been isolated from endothelial cells of many organs. Its localization within endothelial cells has not been determined, nor the mechanisms by which it might be released from endothelial cells during normal organogenesis. METHODS: Localization of FGF within endothelial cells of the anterior pituitary was accomplished by immunocytochemistry and studied by light- and electron microscopy. Capillaries within the anterior pituitary were studied in fetal rats from day 15 to term, and in adult rats. RESULTS: At the onset stages of vascularization (15-18 days fetal), the cytoplasm of the endothelial cells of many of the invading, immature capillaries (thick-walled with few or no fenestrations) was intensely immunopositive for FGF. Immunoprecipitate-filled blebs and slender cytoplasmic processes projected from the endothelial cells into the presumptive pericapillary space and toward the parenchymal cells. As gestation progressed (19-20 day fetal), and an increasing number of capillaries acquired the features characteristic of capillaries in the anterior pituitary of adult animals, i.e., thin-walled and fenestrated, there were fewer capillaries demonstrating immunopositivity for FGF. Foci of released FGF, i.e., extracellular, were occasionally evident within the presumptive pericapillary spaces throughout gestation. By comparison, capillaries of the anterior pituitary of adult rats did not contain immunostainable FGF in their cytoplasm, nor were any blebs and/or processes filled with immunoprecipitate evident. However capillaries did reveal an immunopositive enhancement of their lumenal and ablumenal surfaces. CONCLUSIONS: During vascularization of the anterior pituitary, FGF within the cytoplasm of endothelial cells is released from blebs and/or processes of endothelial cells, and after the capillary bed is stabilized postnatally, these characteristics of vascularization are absent. PMID- 8731040 TI - Tumor necrosis factor alpha (rhTNF) fails to stimulate angiogenesis in the rabbit cornea. AB - BACKGROUND: The objective of this study was to thoroughly examine the in vivo angiogenesis activity of human recombinant tumor necrosis factor alpha (rhTNF). METHODS: rhTNF (0.5 ng to 1.0 microgram) was incorporated into the slow release polymers Hydron or HYPAN and implanted into the rabbit cornea. Release of biologically active rhTNF from the polymers was determined with the L929 cytotoxicity assay. RESULTS: All concentrations tested failed to elicit capillary formation beyond that observed for controls. Less than 2% of the rhTNF was released from the Hydron over 7 days. HYPAN released five times the amount of rhTNF in vitro, but even at doses of 500 ng (104.3 ng suggested release) no angiogenesis was stimulated. CONCLUSIONS: Under the circumstances tested, rhTNF is not angiogenic in vivo. PMID- 8731041 TI - Cytoskeletal differences between stereocilia of the human sperm passageway and microvilli/stereocilia in other locations. AB - BACKGROUND: Stereocilia of the human ductus epididymidis and ductus deferens display unique features in that they arise from an apical cell protrusion (hillock) and contain thick stem portions which are interconnected by cytoplasmic bridges. The molecular basis for this unique fusion and branching pattern is hitherto unknown. These morphologic specialties led us to study the cytoskeleton of male spermway stereocilia with respect to the major proteins that constitute the supportive cytoskeleton of intestinal microvilli and inner ear stereocilia. METHODS: Samples of the human epididymidis and ductus deferens were studied by immunoblotting and immunocytochemistry at the light and electron microscope levels. RESULTS: Spermway stereocilia are supported by an internal actin filament bundle crosslinked by fimbrin and associated with the membrane linker molecule ezrin. The stem portions and hillock area are supplied with the crossbridge forming molecule alpha-actinin. Spermway stereocilia differ from brush border microvilli of the intestine, kidney, and ductuli efferentes by the lack of the second bundling protein villin and the unusual expression of alpha-actinin in the stem region. They resemble inner ear stereocilia by the presence of fimbrin and absence of villin, but differ from them by expression of ezrin and alpha-actinin. Thus, the main molecular difference between spermway stereocilia and stereocilia/microvilli of other locations is the presence of alpha-actinin in their stem portion and the hillock area. CONCLUSIONS: Since alpha-actinin can form crossbridges between adjacent actin filaments (bundles) at longer distances than the other crosslinker of the stereocilium core bundle, fimbrin, we assume that alpha-actinin is essential for both the formation of the stem portions of spermway stereocilia and for the generation of their striking branching pattern. A developmentally regulated temporal sequence of expression of fimbrin and alpha actinin might control the unique architecture of spermway stereocilia. PMID- 8731042 TI - Giant mitochondria induced in rat pancreatic exocrine cells by ethanol and iron. AB - BACKGROUND: Mitochondria are sensitive indicators of cellular pathology. Under certain circumstances, these organelles respond to cellular insult by a marked increase in size, resulting in the formation of giant mitochondria (megamitochondria). Ethanol has been implicated in the induction of giant mitochondria in rat hepatocytes. Since ethanol is reported to affect pancreatic mitochondria, we examined this organ for evidence of mitochondrial giantism in rats administered ethanol and a relatively small amount of supplementary iron. METHODS: Diets were administered via a chronically implanted gastrostomy catheter. Rats were segregated in four groups: 1) basic high-fat diet, 2) ethanol and a high fat diet, 3) carbonyl iron and a high-fat diet, and 4) ethanol and carbonyl iron combined with a high-fat diet. After the animals were on their respective diets for 16 weeks, specimens of pancreas were extirpated and processed for and examined by transmission electron microscopy. RESULTS: Mitochondria in rats on the basic high fat diet resemble those in untreated controls. With ethanol and a high-fat diet, some mitochondria in virtually every exocrine cell are profoundly altered. Such organelles, which are of normal size, have undergone rearrangement of their internal membranes, with three of four parallel cristae residing at one or both poles of spherical to ovate mitochondria. In rats receiving both ethanol and carbonyl iron, giant mitochondria are present in pancreatic exocrine cells. Except for their size, megamitochondria in many cases are virtual likenesses of the altered mitochondria in the alcohol-high-fat rats, having several stacked cristae at either pole and an enhanced matrix compartment. Many of the giant mitochondria have at least one expanded crista, which contains packets of helically coiled filaments. When control rats are fed carbonyl iron without ethanol, the pancreatic mitochondria display a marked propensity for forming clusters of tightly interlocked organelles, formations that may be a prelude to mitochondrial fusion. CONCLUSIONS: It appears that iron supplementation sets the stage for organelle fusion, hence enlargement, with the ethanol providing the stimulus for fusion to actually take place, and controlling the final morphology of the resultant megamitochondria. PMID- 8731043 TI - Hypoplasia of cushion ridges in the proximal outflow tract elicits formation of a right ventricle-to-aortic route in retinoic acid-induced complete transposition of the great arteries in the mouse: scanning electron microscopic observations of corrosion cast models. AB - BACKGROUND: The major morphologic change associated with retinoic acid (RA) induced complete transposition of the great arteries (TGA), a congenital malformation of the heart, was investigated in a mouse model in which TGA was found in 80% of surviving fetuses. METHODS: Corrosion casts of embryonic hearts with or without prior exposure to retinoic acid were observed under a scanning electron microscope. RESULTS: In control hearts, indentations caused by expanded parietal and septal ridges in the outflow tract established right ventricle-to left ventral pulmonic and left ventricle-to-right dorsal aortic routes before the aorticopulmonary septum completion. In RA-treated hearts, indentations of proximal regions of the parietal and septal ridges were small in the proximal outflow tract, whereas those in the distal regions developed well. These morphological features in the RA-treated hearts elicited right ventricle-to-right ventral aortic and left ventricle-to-left dorsal pulmonic routes in the TGA morphology. CONCLUSIONS: Hypoplasticity of the proximal regions of parietal and septal ridges in the outflow tract is one of the primary morphological abnormalities of the RA-induced TGA. PMID- 8731044 TI - Local and regional variations in myofibrillar patterns in looping rat hearts. AB - BACKGROUND: In chickens, cytodifferentiation, right side dominance in myofibril development, and variations in myofibrillar patterns in different areas and layers of the myocardial wall exist which have been implicated in the process of heart looping. Little comparable information is available for developing myofibrillar patterns in the early development of mammalian hearts. METHODS: We have used transmission electron microscopy (TEM), confocal scanning laser microscopy (CSLM), and 3-D reconstruction techniques also present in the looping hearts of embryonic day (ED) 9.5 to 11.5 rat hearts. RESULTS: Local and regional variations and right side dominance in myofibrillar patterns were shown during looping in 9.5 through 11.5 days of development in embryonic rat heart. At 9.5 days of development, myofibrils near the lumen of the myocardial wall were primarily in circumferential bands while near the pericardial surface they were primarily in longitudinal bands. In older embryos, regional variations in myofibrillar organization was found in areas associated with the cardiac cushions, trabeculae, and myocardial wall of the developing heart chambers. Based on sarcomeric structure, myofibrils in the ventricle and outflow tract were more advanced than those found in the atrial wall. CONCLUSIONS: The local and regional patterns of myofibrils in looping rat hearts are similar to those which have been found in developing chicken hearts. This study and others indicate cytodifferentiation and development of the contractile apparatus has a crucial role in the process of heart looping. PMID- 8731046 TI - Tonsils of the soft palate of young pigs: crypt structure and lymphoepithelium. AB - BACKGROUND: Tonsils of the soft palate are especially important in pigs as the major pharyngeal mucosal-associated lymphoid tissue and as a portal of entry for microorganisms. They play a key role in initiating immune responses against antigens entering tonsillar crypts from the oropharynx. The aim of this study was to describe the architecture of the tonsillar crypst and the morphology of their epithelial surface. METHODS: Tissue taken from the tonsil of the soft palate of freshly-killed pigs was examined using light microscopy, electron microscopy, and three-dimensional reconstruction techniques. RESULTS: Tonsils of the soft palate in pigs are penetrated by numerous crypts which extend into, and branch extensively within, the lymphoid tissue. Stratified squamous non-keratinised epithelium covering the oropharyngeal surface is continuous with that lining the neck of crypts. Lymphoepithelium covers the tonsillar lymphoid tissue within the crypts. It consists of non-keratinised epithelial cells, M cells, goblet cells and many intraepithelial lymphoid cells. M cells have a variable surface morphology: some are covered by relatively regular and well-formed microvilli; others possess very long undulant microvilli emanating from broad membranous folds. CONCLUSIONS: Variations in M cell surface morphology occur and these may reflect alterations of the apical plasmalemma in response to antigenic stimuli. Further investigation will be required to determine molecular specializations of the apical membrane of M cells which may facilitate interactions with antigenic material. PMID- 8731045 TI - Development of the sympathoadrenal system in the chick embryo: an immunocytochemical study with antibodies to pan-neuroendocrine markers, catecholamine-synthesizing enzymes, proprotein-processing enzymes, and neuropeptides. AB - BACKGROUND: The adrenal chromaffin cells synthesize, store and secrete a complex mixture containing amines, structural proteins, enzymes, and neurohormonal polypeptides. Most of the studies dealing with the development of the avian sympathoadrenal system have been based on antibodies recognizing signal molecules like HNK-1, NC-1, and N-CAM. METHODS: The development of the chick sympathoadrenal system was studied from 3 1/2 to 21 days of incubation, both morphologically and immunocytochemically, using antibodies to 17 separate antigens, including antibodies to pan-neuroendocrine markers, catecholamine synthesizing enzymes, proprotein-processing enzymes, and neuropeptides. RESULTS: Some of the antigens studied were heavily expressed from the first days of development, e.g., chromogranin-A, chromogranin-B, Go protein-alpha subunit, tyrosine hydroxylase, and galanin, while for others a strong heterogeneity both in number of immunoreactive cells and intensity of immunostaining was recorded at the different stages, e.g., dopamine-beta-hydroxylase,, 7B2 protein, proprotein convertase 2, somatostatin, met-enkephalin, secretogranin II, proprotein convertase 3, neuropeptide Y, phenyl-N-methyl transferase, and neuron-specific enolase. The first immunoreactivities to appear at day 3 1/2 were those for HNK 1, tyrosine hydroxylase, chromogranin-A, and chromogranin-B. Except for HNK-1, immunoreactivity for all the remaining antigens showed a steady increase up to the hatching. CONCLUSIONS: Three expression patterns were found, in the developmental adrenal-gland: defining early permanent markers (chromogranin-A, chromogranin-B, Go protein-alpha subunit, tyrosine hydroxylase, and galanin), others that show a progressively increased expression until the day 10 of development (dopamine-beta-hydroxylase, 7B2 protein, proprotein convertase 2, somatostatin, met-enkephalin), and late-appearing antigens (secretogranin II, proprotein convertase 3, neuropeptide Y, phenyl-N-methyl transferase, and neuron specific enolase). PMID- 8731047 TI - Development of the human wrist joint ligaments. AB - BACKGROUND: Many studies have been published on the development of the human wrist joint, but scant attention has been given to the development of the wrist joint ligaments. Moreover, traditional description of wrist anatomy usually depict only the superficial capsular fibers of the wrist joint. The only ligamentous structure to receive much attention is the articular disc of the wrist joint, which has been described as a fibrocartilaginous structure extending from the medial edge of the lower end of the radius to the ulnar styloid process. METHODS: In the present report, we synthesize our observations in the wrist joint ligaments in 35 serially sectioned human embryonic and fetal hands (16 embryos and 19 fetuses). RESULTS: The interosseous intercarpal ligaments are organized from the mesenchyme, which, until O'Rahilly's stage 23, fills the intercarpal spaces. These ligaments are not individually distinguishable until the 9th week of development. The collateral ligaments begin to form in O'Rahillys's stage 22 and are completely formed by the end of week 10. The palmar radiocarpal and ulnocarpal ligaments (beginning with the palmar radiocarpal ligament) begin to form in O'Rahilly's stage 23 and are fully developed by the end of week 10. At this time, development of the dorsal radiocarpal ligament begins; this process is completed by the end of week 13. The articular disc which is initially formed of a single element, first appears in O'Rahilly's stage 23 and its organization is completed at week 10 of development. CONCLUSIONS: We establish the morphogenetic time-table of the wrist joint ligaments. Our descriptive findings may help explain carpal motion and the origin of wrist injuries. PMID- 8731048 TI - Stabilizing role of the basement membrane and dermal fibers during newt limb regeneration. AB - BACKGROUND: Following amputation of a newt limb, tissues at the amputation site undergo histolysis to give rise to a growth bud, or blastema, but they also provide a base on which the regenerate is constructed. Studies suggest that dermal tissues may differentially resist histolysis. METHODS AND RESULTS: To examine stability of tissues at the amputation site, more than 80 preblastemal staged regenerating limbs were examined histologically. Initially, all soft tissues not attached to bone retracted and were covered by migrating epithelium. The dermis was seen to be stable during the first week postamputation. Muscle dedifferentiated and was heavily stained with anti-tenascin antibodies, but the intact overlying dermis was unstained. Fiber bundles, revealed by staining with phosphotungstic acid hematoxylin, isolated the dermis from dedifferentiating deeper tissues during the first week postamputation, but partially broke down during the second week. However, the basement membrane (BM) remained as the distalmost intact structure at the amputation site in all limbs examined. The BM was the foundation for new BM synthesis which preceded dermis synthesis in the base of the blastema during the second week, even while undifferentiated cells were accumulating centrally. CONCLUSIONS: We suggest that the dermis resists histolysis long enough for new BM to form in continuity with that of the stump. Dermis formation (dermogenesis) distal to the amputation plane begins early as in mammalian healing but is not completed until after blastema formation. Thus, factors that inhibit dermal closure appear to distinguish regenerating from non regenerating appendages. PMID- 8731049 TI - Inactivation of brainstem motor nuclei blocks expression but not acquisition of the rabbit's classically conditioned eyeblink response. AB - Rabbits were eyeblink conditioned while their accessory abducens nucleus (ACC), facial nucleus (FN), and surrounding reticular formation (RF) were temporarily inactivated with microinjections of muscimol to determine whether these structures are critically involved in acquisition of the conditioned eyeblink response (CR). Rabbits performed no CRs or unconditioned responses (URs) during inactivation training. Training was continued without inactivation and rabbits performed the CR at asymptotic levels from the start of training without inactivation. They had fully learned the CR while their ACC, FN, and RF were inactivated, despite performing no CRs or URs at all during inactivation. These results rule out any critical role for neurons within the ACC, FN, or surrounding RF in acquisition of the classically conditioned eyeblink response. PMID- 8731050 TI - Amygdala central nucleus lesions attenuate acoustic startle stimulus-evoked heart rate changes in rats. AB - Amygdala central nucleus (CNA) lesions were used to test the hypothesis that stimulus-evoked heart rate changes can reflect the development of fear during acoustic startle testing. A 120-dB white noise startle stimulus produced freezing as well as phasic heart rate accelerations and decelerations, and an abrupt decrease in tonic heart rate, in sham-operated rats. These responses were all significantly reduced in CNA-lesioned rats. In contrast, an 87-dB stimulus elicited only significant phasic decelerations that were similarly attenuated by the CNA lesions. In a follow-up experiment, the CNA lesions also attenuated phasic cardiac decelerations evoked by a conditioned stimulus-like, 85-dB pure tone. The results support the contention (B. J. Young & R.N. Leaton, 1994) that heart rate changes can reflect fear conditioned during acoustic startle testing and, in addition, suggest that the amygdala mediates responses to nonsignal acoustic stimuli. PMID- 8731051 TI - Postconditioning isolation disrupts contextual conditioning: an experimental analysis. AB - Contextual-fear conditioning requires a lengthy retention period to fully emerge. This phenomenon might reflect the consolidation of a representation of the context that can be used to evoke fear. To investigate this hypothesis, 25 day old rats that were returned to their home cages after conditioning were compared with rats that were isolated in a novel room. Isolation disrupted contextual but not auditory-cue fear conditioning when the conditioning-isolation interval was 2 hr or less, but not when it was 24 hr. Preexposure to the context prevented the isolation effect, and isolation disrupted this effect of context preexposure. These results support the consolidation hypothesis and the view that contextual- and auditory-cue fear conditioning depend on different processes. PMID- 8731052 TI - Behavioral vigilance following infusions of 192 IgG-saporin into the basal forebrain: selectivity of the behavioral impairment and relation to cortical AChE positive fiber density. AB - Rats were trained in a previously validated behavioral vigilance task that required them to detect visual signals of variable length and to discriminate signal from nonsignal events. Baseline performance was characterized by a signal length-dependent ability to score hits, a decline in hits over time, and a correct rejection rate of approximately 70%. After the rats reached criterion performance in this task, the immunotoxin 192 IgG-saporin or its vehicle was infused into the area of the nucleus basalis/substantia innominata of the basal forebrain. Postoperative performance in lesioned rats was characterized by a decrease in their ability to detect signals while their ability to correctly reject nonsignals remained unaffected. The effect of the lesion did not recover in the course of over 180 sessions of postlesion testing. The overall performance of the rats correlated with acetylcholinesterase (AChE)-positive fiber density in all cortical areas measured except the cingulate and pyriform cortex. These findings help to elucidate the nature of the attentional impairments resulting from the loss of cortical cholinergic inputs. PMID- 8731053 TI - Ischemia-induced object-recognition deficits in rats are attenuated by hippocampal ablation before or soon after ischemia. AB - The literature on the role of the hippocampus in object-recognition contains a paradox: Transient forebrain ischemia (ISC) produces hippocampal damage and severe deficits on the delayed nonmatching-to-sample (DNMS) task, yet hippocampal ablation (ABL) produces milder deficits. Experiment 1 confirmed that pretrained rats display severe DNMS deficits following ISC, but not ABL. Ischemia produced loss of CA1 neurons, but no obvious extrahippocampal damage. In Experiments 2 and 3, ISC rats from Experiment 1 received ABL, and ABL rats received ISC; neither treatment affected DNMS performance. In Experiment 4, rats that received ISC followed 1 hr later by ABL displayed only mild deficits. It is hypothesized that ISC-induced DNMS deficits are due to extrahippocampal damage produced by pathogenic processes that involve the hippocampus. PMID- 8731054 TI - Contralateral turning caused by metabotropic glutamate receptor stimulation in the dorsal striatum is reversed by MCPG, TTX, and cis-flupenthixol. AB - Recent evidence suggests an involvement of metabotropic glutamate receptors in the physiology of the striatum. In this study, rotation was recorded in an automated rotometer for 20 min following dorsal striatal injections (0.5 microliter) in cannulated rats. The metabotropic agonist 1-aminocyclopentane trans-1, 3-dicarboxylic acid (1S,3R-ACPD) caused dose-dependent contralateral rotation. Turning caused by 500 microM 1S,3R-ACPD was reversed by coinjections of the metabotropic antagonist (+)-alpha-methyl-4-carboxyphenylglycine (MCPG, 1 mM) and by tetrodotoxin (100 microM). Injections of MCPG alone (10 microM, 100 microM, 1 mM) failed to elicit turning. Increasing doses of the dopamine antagonist cis-flupenthixol also reversed 1S,3R-ACPD-induced rotation. Thus unilateral striatal metabotropic glutamate receptor stimulation can cause receptor-specific rotation that may result from an increase in neural activity, and is dependent on intact dopamine neurotransmission. PMID- 8731055 TI - Intraocular transfer and simultaneous processing of stimuli presented in different visual fields of the pigeon. AB - Differences in neuroanatomy, optics, and function indicate the operation of 2 visual systems in pigeons, a frontal field system and a lateral field system. Communication between these systems was examined with a delayed matching-to sample task in which sample stimuli could be presented in either the frontal or lateral fields. In Experiment 1, matching acquired with the lateral field transferred to the frontal field but did not transfer from the frontal field to the lateral field. When different samples were presented simultaneously to the frontal and lateral fields in Experiment 2, pigeons preferred to match the sample in the frontal field, but lateral field information interfered to some extent with frontal field matching. The 3rd experiment showed left lateral field dominance when the left and right fields were simultaneously presented with different sample stimuli; left field dominance was not complete, as pigeons sometimes matched the right-field sample. PMID- 8731056 TI - The NMDA antagonist MK-801 affects nonspatial learning in preweanling rats. AB - The N-methyl-D-aspartate (NMDA) subtype of the excitatory amino acid receptor has been implicated in several kinds of learning and memory, as well as in long-term potentiation (LTP), a putative cellular mechanism for learning and memory. This experiment examined the role of the NMDA receptor in patterned single-alternation (PSA) learning in preweanling rats following intraperitoneal injections of 0.05 mg/kg MK-801, a selective NMDA antagonist. MK-801 significantly inhibited PSA at both 60-s and 30-s intertrial intervals (ITIs), and attenuated, but did not block, learning at 8-s ITI. These results are compared with effects on PSA, a form of nonspatial, memory-based learning, observed after early postnatal exposure to alcohol, infant hippocampal lesions, and infant exposure to X irradiation, and they add strongly to these earlier demonstrations of the role of the hippocampus in learning and memory that is clearly nonspatial and non cognitive-map-related. PMID- 8731058 TI - Cocaine transiently impairs maternal behavior in the rat. AB - This paradigm distinguished between two hypotheses not previously directly addressed. Do repeated exposures to cocaine at critical times during pregnancy, when the neural mechanisms that support maternal behavior are being read, alter some fundamental neural underpinning of maternal behavior in rats? Alternatively, does cocaine alter maternal behavior only when circulating? During the 4 hr after cocaine injection (20 or 40 mg/kg), there were significant deficits in maternal behavior. In contrast, 16 hr after cocaine injection, drug-injected females, in which plasma cocaine had fallen to nondetectable levels, showed the normal maternal behavior of saline-injected controls. This pattern of impaired maternal behavior after cocaine injection, followed by normal behavior as blood levels returned to zero, was replicated over 8 days. It was concluded that cocaine impairs maternal behavior only when circulating and does not have a residual effect in the transiently drug-free, chronically drug-treated dam. PMID- 8731057 TI - Thermogenesis, myoclonic twitching, and ultrasonic vocalization in neonatal rats during moderate and extreme cold exposure. AB - Physiological and behavioral responses of 2- and 7-8-day-old rats were monitored during moderate and extreme cold exposure. During moderate cold exposure (30 degrees C < or = air temperature < or = 32.5 degrees C), pups at both ages increased heat production, maintained an elevated interscapular temperature, and maintained baseline levels of myoclonic twitching, a behavior commonly associated with active sleep. During extreme cold exposure (21 degrees C < or = air temperature < or = 25 degrees C), pups at both ages continued producing metabolic heat, but now exhibited pronounced decreases in interscapular temperature and decreased rates of myoclonic twitching. Furthermore, the 7-8-day-old pups exhibited significant increases in ultrasound production, and males vocalized more than females. These results suggest the presence of a narrow subthermoneutral zone in neonates in which nonshivering thermogenesis is regulated and sleep-related behaviors are protected. PMID- 8731059 TI - The isolation and companion comfort responses of 7- and 3-day-old rat pups are modulated by drugs active at the opioid receptor. AB - Rat pups emit ultrasonic vocalizations (USVs) when isolated in a novel environment. In 10-day-olds, USV has been shown to be reduced by either the administration of 0.125 mg/kg morphine (MOR) or the presence of a littermate; these effects were both reversed by naltrexone (NLX), an opioid receptor blocker. The present study reports that the same dose of MOR produced NLX-antagonized quieting without sedation in 7- and 3-day-old pups; higher doses of MOR decreased USV but produced motor deficits as well. The 0.125 mg/kg dose of MOR is less effective in reducing USV in 3- and 7-day-olds; calling rates declined by no more than 42%, compared with 65% at 10 days of age. The presence of a companion also lowered the USV of 3- and 7-day-olds by a lesser amount (55-57%) than the 67% seen in 10-day-olds or the 90% decline when pups are 2 weeks old. This suggests that age-related changes in the opioid system may be relevant to the increased salience of a social companion that comes with maturity. PMID- 8731060 TI - Dissociations between appetitive and consummatory responses by pharmacological manipulations of reward-relevant brain regions. AB - Appetitive behaviors of rats were monitored in a runway situation following central infusions of neuroactive substances into brain areas implicated in electrical self-stimulation. Microinjections of the dopamine antagonist cis flupentixol or the cholinergic antagonist atropine into the nucleus accumbens (Acb) severely reduced the approach speed and anticipatory shuttlebox activity while leaving the consumption of the 20% sucrose reward intact. Microinjections of GABA into the ventral tegmental area (VTA), pedunculopontine tegmental nucleus (PPTg), and oral pontine reticular nucleus (PnO) also severely disrupted approach without decreasing consumption. The highest doses of atropine into the VTA, PPTg, and PnO disrupted both consummatory and approach responses equally. The results indicate that modulation of various neurochemistries along the trajectory of the self-stimulation system has stronger effects on appetitive approach than consummatory motivation. The implications for understanding appetitive-approach motivation in the brain are discussed. PMID- 8731061 TI - Shuttle-box memory facilitation by posttraining intracranial self-stimulation: differential effects in rats with high and low basic conditioning levels. AB - The effects of intracranial self-stimulation (ICSS) on retention (after 24 hr, 7, 15, or 60 days) of a massed 2-way active avoidance task were studied in independent groups of rats. All groups showed a higher performance on the retention session than on the acquisition one. In the control subjects, the higher retention performances were observed in the 7- and 15-day groups. However, the ICSS treatment facilitated the 24-hr retention compared with its control group, allowing the treated subjects to achieve the same level of performance on the 24-hr retention session than that achieved by the control rats at the 7-day retention test. In the 24-hr groups, the facilitatory ICSS effect was stronger in the subjects with a low level of conditioning and weaker in those with a high level. Results suggest that posttraining ICSS accelerates memory consolidation and equalizes the performance of poor and good learners. PMID- 8731062 TI - The physiology of social conflict in rats: what is particularly stressful? AB - The involvement of the opponent's behavior in the aggression-induced stress response was tested. Subjects (male Wistar rats) faced stimulus rats previously submitted either to mediobasal hypothalamic lesion (MBHL) or to sham operation. MBHL intruders were more aggressive but were also more ready to submit compared with sham-operated rats; as residents they were more efficient in inducing submissive behavior in the opponent. The subject's stress response at 15 min was neither dependent on the intensity of aggressive behavior nor on residence status but on dominant-submissive relationships. The submission of the stimulus rat reduced corticosterone plasma levels in the subjects, while the subjects' own submissive behavior elicited a very significant increase in its plasma corticosterone. In contrast, plasma glucose seemed to depend on residence status, whereas plasma free fatty acids reacted to any novel event (including cage switch). PMID- 8731063 TI - Stress controllability influences the ataxic properties of both ethanol and midazolam in the rat. AB - Rats were administered either 80 escapable shocks or yoked inescapable shocks, were then injected with saline or several ataxic doses of either ethanol or midazolam, and then had their motoric impairment assessed by Rotarod performance. No motoric impairment was observed following saline injection. However, inescapable shock impaired Rotarod performance in response to both ethanol and midazolam at 2 hr, but not immediately poststress. Conversely, escapable shock reduced the ataxic potency of ethanol, although it had no influence on midazolam induced ataxia. These results indicate functional alterations in behavioral reactivity to low doses of several classes of central nervous system depressants by psychological dynamics of stress exposure. Our findings demonstrate the impact of stress controllability on behavioral reactivity to two classes of drugs of abuse. PMID- 8731065 TI - Clonidine antagonizes the aversive effects of opiate withdrawal and the rewarding effects of morphine only in opiate withdrawn rats. AB - The researchers asked whether clonidine, an alpha 2-noradrenergic agonist, would block selectively the motivational effects of opiate withdrawal and whether clonidine's effects would respect the boundary between nondeprived and deprived motivational states. In a place conditioning paradigm, clonidine (0.05 mg/kg ip) blocked the rewarding effects of morphine in opiate-withdrawn rats (as well as the aversive properties of withdrawal itself), but did not affect morphine place preferences (2 and 20 mg/kg) in drug-naive rats. Furthermore, clonidine blocked the acquisition of morphine (15 mg/kg), but not LiCl (15 mg/kg), conditioned taste aversions in water-deprived rats. The results suggest that the motivational system activated in deprived animals includes dopaminergic and noradrenergic components that are in series with each other. PMID- 8731064 TI - The differential contribution of spinopetal projections to increases in vocalization and motor reflex thresholds generated by the microinjection of morphine into the periaqueductal gray. AB - The capacity of morphine microinjected into the ventrolateral periaqueductal gray (vPAG) to elevate the thresholds of spinal motor reflexes (SMRs), vocalizations during shock (VDSs) and vocalization afterdischarges (VADs) was challenged by the intrathecal administration of receptor antagonists to serotonin (methysergide), norepinephrine (phentolamine) and mu-opiates (naloxone). Methysergide and phentolamine were equipotent in reversing increases in SMR thresholds. The efficacy of these antagonists to reduce increases in VDS and VAD thresholds was dependent on the dose of morphine administered into the vPAG. These results indicate that the dose of morphine administered into the vPAG determines the contribution of spinopetal projections in inhibiting dorsal horn neurons involved in reflex generation versus the rostral transmission of pain information. A hypothesis is offered regarding the mechanisms by which vPAG administered morphine suppresses nociceptive transmission through different levels of the neuraxis. PMID- 8731066 TI - Brief, high-frequency stimulation of the corticomedial amygdala induces a delayed and prolonged increase of aggressiveness in male Syrian golden hamsters. AB - Brief 200-Hz stimulation of the corticomedial amygdala (CMA) increases the aggressiveness of male Syrian golden hamsters for about 30 min; the effect peaks 10-15 min after stimulation. This effect is sensitive to stimulation amplitude and frequency. Stimulation at the parameters that reduce attack latency increases flank marking but does not affect copulation latency or general activity. Immunocytochemical analysis suggests that stimulation effects may be coupled to c fos expression and that unilateral stimulation has bilateral effects. CMA stimulation effects appear to mimic part of the time course of behaviorally induced attack priming. The temporal persistence of aggression may result from long-term potentiation-like changes within CMA-related neural circuitry. PMID- 8731067 TI - Effects of perinatal testosterone on handedness of gerbils: support for part of the Geschwind-Galaburda hypothesis. AB - When assuming their species-typical tripodal stance, male Mongolian gerbils (Meriones unguiculatus) use their right forepaw for support more frequently than do females. This experiment determined whether, as N. Geschwind and A. M. Galaburda (1987) have proposed, the direction of such sexually dimorphic lateral asymmetry of forelimb use is affected by perinatal exposure to testosterone (TP). It was found that male gerbils injected with TP when 4 days old were significantly less likely to rest on their right forepaw when in a tripodal stance than were their oil-injected siblings. Female gerbils injected with TP when 6 days old were more likely than oil-injected controls to use their right forepaws for support. The findings demonstrate effects of perinatal exposure to TP on handedness in gerbils and suggest that the relationship between TP exposure and asymmetrical forelimb use is not always as direct as Geschwind and Galaburda's model suggests. PMID- 8731068 TI - Deletion of leukocyte mitochondrial DNA in bipolar disorder. AB - Leukocyte mitochondrial DNA (mtDNA) was examined in 35 patients with bipolar disorder by the nested PCR method to explore whether or not the 4977 base-pair deletion (common deletion) is found. The PCR product corresponding to the common deletion was found in 2 of 35 (5.7%) patients and none of 29 normal controls. It was confirmed by the primer shift PCR method that this PCR product was amplified from deleted mtDNA. These results suggest that more than a small percentage of patients with bipolar disorder might have deleted mtDNA and that this aberrant mtDNA might relate to pathophysiology of a subtype of bipolar disorder. PMID- 8731069 TI - Personality dimensions in panic disorder and generalized anxiety disorder. AB - To make a dimensional assessment of personality in individuals with pathological anxiety, the Tridimensional Personality Questionnaire (TPQ) was administered to 32 patients with panic disorder (PD) and 49 patients with generalized anxiety disorder (GAD). The most striking findings were a substantially increased score on the harm avoidance dimension in both groups of patients, and a lack of significant differences between the TPQ scores in patients with PD and GAD. The former finding suggests that higher levels of harm avoidance may be common to (although not necessarily specific for) various types of anxiety disorders. The latter finding is in agreement with the findings that PD and GAD do not differ significantly with respect to the associated personality disorder diagnoses, which may further cast a doubt on the validity of the distinction between PD and GAD. PMID- 8731070 TI - Dependency, matching adversities, length of survival and relapse in major depression. AB - This paper reports the 1-year follow-up of a sample of depressed patients. Social cognitive variables obtained during the index episode, including ideal emotional support, roles and goals investment in various domains and dysfunctional attitudes were used to predict subjects' likelihood for subsequent relapse. More subjects who experienced severe life events in the year were found to have relapsed. However, the predictive value of life events was improved if adversity was in the most invested domain according to the roles and goals questionnaire that subjects filled in during their index episode (matching adversity). Subjects who experienced matching adversity had a 3-fold chance of relapse compared with subjects with nonmatching adversity. The majority of the matching events were in the interpersonal domain. Levels of dysfunctional attitudes alone did not predict relapse. However, matching adversity and the dependency subscale of the dysfunctional attitude scale contributed significantly both to whether or not subjects relapsed and to the number of weeks subjects survived before they relapsed. The higher the level of dependency dysfunctional attitudes, the sooner subjects relapsed. The findings of the follow-up study supported the importance of psychological and social factors in determining relapse. PMID- 8731071 TI - Social adjustment in dysthymia, double depression and episodic major depression. AB - We contrasted the overall social functioning of pure dysthymics, double depressives, episodic major depressives and normal controls using both interview and self-report measures of social functioning and depression. In addition, we used hierarchical multiple regression to assess the differential impact of several variables (comorbid personality, anxiety and substance use disorders, life stress, duration of dysthymia and severity of depressive symptomatology) on social functioning in the dysthymics and double depressives. Participants included 41 outpatients with early-onset dysthymia alone, 56 outpatients with early-onset dysthymia and concurrent major depression, 45 outpatients with episodic major depression and 45 normal controls. All 3 patient groups were found to be significantly more impaired than normal controls in overall functioning, as well as in every specific role area. Double depression was found to be particularly impairing, both in overall functioning and in every specific role area. In dysthymic patients with and without concurrent major depression, current depressive symptomatology is the strongest predictor of impairment. Taken together, these data suggest that chronic, low-grade depressive symptoms and acute, moderate depressive symptoms have similar, significant and additive effects on social adjustment. PMID- 8731072 TI - Low levels of MHPG in depressive spectrum patients. AB - There is some preliminary laboratory support for the proposed classification of depressed patients into depressive spectrum disease (DSD) and non-DSD. This study explores whether there is a difference in the levels of the norepinephrine metabolite, MHPG, in DSD and non-DSD patients. MHPG levels from 38 DSD patients were compared with 24 non-DSD patients. After controlling for the influence of age and gender on MHPG, the DSD patients had MHPG levels that were lower than non DSD patients; 1655 +/- 90 mg/day vs. 1965 +/- 174 mg/day, respectively; P = 0.05. This study provides additional laboratory support for the DSD subtype. Possible implications of this finding are discussed. PMID- 8731073 TI - 'Natural' light treatment of seasonal affective disorder. AB - Patients with seasonal affective disorder (SAD) were treated for 1 week either with a daily 1-h morning walk outdoors (natural light) or low-dose artificial light (0.5 h@2800 lux). The latter treatment (given under double-blind conditions) can be considered mainly placebo and did not improve any of the depression self-ratings, whereas natural light exposure improved all self ratings. According to the Hamilton depression score, 25% remitted after low-dose artificial light and 50% after the walk. Sleep duration or timing were not crucial for the therapeutic response. The morning walk phase-advanced the onset and/or offset of salivary melatonin secretion, but individual clinical improvement could not be correlated with specific phase-shifts. Morning cortisol was decreased. Low-dose artificial light did not modify melatonin or cortisol patterns. This is the first study to provide evidence for the use of outdoor light exposure as a potential alternative or adjuvant to conventional artificial light therapy in SAD. PMID- 8731074 TI - Advanced vs. normal sleep timing: effects on depressed mood after response to sleep deprivation in patients with a major depressive disorder. AB - Total sleep deprivation (TSD) exerts beneficial but only transient effects on mood in patients with a major depressive disorder (MDD). Though approximately 50 to 70% of depressed patients improve after sleep deprivation, the majority relapse after recovery sleep, some even after a short nap. One theoretical model postulates a critical period in the early morning hours where sleep is likely to induce a relapse, and nap studies indicate that sleep may be particularly 'depressogenic' at this time of day. A second model attributes the relapse to the release of non-REM sleep. We therefore compared the impact of an advanced sleep period (17:00-24:00 h) to a normal sleep period (23:00-06:00 h) on mood in patients who had responded to sleep deprivation. Less relapses into depression occurred after advanced sleep. Polysomnographic data showed that, as expected, normal sleep was characterized by a more pronounced improvement of sleep continuity and increased slow-wave sleep. The normal sleep group showed a stronger decrease in REM sleep density than the advanced sleep group compared with baseline. These data add to a growing body of evidence that the timing of sleep following successful sleep deprivation may be crucial for a stabilization of its antidepressant effect. Thus, avoidance of sleep during a "critical period' for more than a single night is necessary to provide a longer-lasting treatment modality. PMID- 8731075 TI - Seasonal affective disorder among psychiatric nurses in Aberdeen. AB - The Seasonal Pattern Assessment Questionnaire was posted to psychiatric nurses in Aberdeen, and 443 (73% of eligible subjects) responded. The rate of Seasonal Affective Disorder (SAD) was 2.9% and for subsyndromal SAD (S-SAD) was 9.5%. When rates were age and sex adjusted to the Aberdeen population aged 15 to 64, this yielded a prevalence of 1.9% for SAD and 7.2% for S-SAD. As in other studies, there was a preponderance of young female sufferers, but being married appeared to be protective. Comparisons with epidemiological studies elsewhere lead to the conclusion that population rates of winter depression relate to a combination of genetic susceptibility/selection, latitude of residence and geographical mobility. PMID- 8731076 TI - Urinary phenylethylamine and cortisol levels in the early puerperium. AB - Phenylethylamine and cortisol were measured in 24-h urine samples taken from women day 2/3 postpartum, and matched controls. They also completed self-rating blues and highs scales. There was no significant difference in phenylethylamine excretion in those who scored highly with blues or highs, and either postpartum or normal controls. The number of raised values for phenylethylamine output was significantly greater in the postpartum women, who had no psychopathology, than in normal controls. Cortisol levels were significantly raised in postpartum controls compared with normal controls; women with the highs excreted significantly less cortisol than other postpartum women. PMID- 8731077 TI - Comparative validity of two measures of psychomotor function in patients with severe depression. AB - Psychomotor function is a key construct in depressed patients and two measures have been developed for systematic rating. Parker and colleagues utilise an observer-rated scale (CORE) while the scale (DRRS) developed by Widlocher assesses motor and ideational aspects. Association between the scales and their relative capacity to predict ECT response were investigated in a sample of 81 depressed patients. Both predicted ECT response. While the CORE scale rates a wider variety of phenomena (including non-interactiveness and agitation) and does not rely on the subject's capacity to report aspects of their cognitive function, the study supports the predictive and comparitive validity of both scales. PMID- 8731078 TI - The influence of climate on development of winter depression. AB - 126 patients suffering from winter depression participated in the study. Every second week patients completed the 13 item Beck Depression Inventory (BDI) in the period September to May in the years 1991 to 1994. Local weather data from this period were obtained from the Meteorological Institute, Copenhagen. No significant correlation was found between score on BDI and cloud cover, rainfall or atmospheric pressure. A significant correlation was found between score on BDI and minutes of sunshine, global radiation, length of daylight and temperature. This is in accordance with the theory, that lack of light is a contributing factor for development of winter depression. PMID- 8731079 TI - Comorbidity of personality disorders and unipolar major depression: a review. AB - The association of major depressive disorders with personality disorders is relevant in terms of clinical, therapeutic and prognostic aspects. However, the prevalence of this association remains unclear. This may be due to methodological considerations. Nonetheless, it could be estimated from this review that 20% to 50% of inpatients and 50% to 85% of out-patients with a current major depressive disorder have an associated personality disorder. Cluster B personality disorders, in particular borderline (10-30%), histrionic (2-20%) and antisocial (0-10%), seem to be overrepresented, even if the narcissistic one is rare (less than 5%). The main characteristic of Cluster C personality disorders is the great variability of results across studies, except for the obsessive compulsive personality disorder, whose prevalence is consistent and rather high (0-20%). Cluster A personality disorders are an heterogeneous group, since the prevalence of schizotypal personality disorder is rather high (0-20%), the prevalence of paranoid personality disorder is low (less than 5%) and the prevalence of schizoid personality disorder is quite variable from one study to another. The prevalence of personality disorders among patients with a lifetime major depression has been insufficiently studied, although it may concern half of these patients. The prevalence of current of lifetime major depression among patients with a personality disorder has not been sufficiently studied and results are very scattered. Also, the coexistence of personality disorder and major depression is frequent, and this review emphasizes the heterogeneity of the personality styles associated with major depression. Finally, an optimization of methods and the adjunction of a dimensional point of view to the categorical approach may help to study the comorbidity of major depression and personality disorders and its consequences. PMID- 8731080 TI - Commerce in transplantation in Third World countries. PMID- 8731081 TI - Multiple intracellular MAP kinase signaling cascades. PMID- 8731083 TI - Localization of authentic thromboxane A2/prostaglandin H2 receptor in the rat kidney. AB - Using a polyclonal antibody against authentic thromboxane A2/prostaglandin H2 (TxA2/PGH2) receptor protein, we assessed the distribution of this receptor in the normal rat kidney by routine methods of immunofluorescence microscopy. The receptor localized both in glomeruli and in tubules. In the former, the distribution of the receptor was most prominent along the lumen of glomerular capillary loops. Parietal epithelial cells of the Bowman's capsule, podocytes and mesangial cells also demonstrated immunostainable receptor. In the tubules, the receptor localized most prominently at the base of the brush border of proximal tubules and at the luminal surface of thick ascending limbs and distal convoluted tubules. These observations point to sites that are likely to be targeted by thromboxane A2 in forms of renal injury characterized by enhanced synthesis of this eicosanoid. PMID- 8731082 TI - Role of adenosine in the renal responses to contrast medium. AB - Despite the development of non-ionic radiographic contrast media (CM), CM-induced nephropathy is a clinically important problem in patients with pre-existing renal insufficiency. We examined the effects of non-ionic CM (iohexol) on renal function in conscious dogs with and without renal insufficiency, and evaluated the effects of a non-selective (theophylline), an A1 selective (KW-3902), and an A2 selective adenosine antagonist (KF17837) on the renal responses to CM. In sham operated group, iohexol (2 ml/kg/min for 3 min) increased effective renal plasma flow (ERPF) and glomerular filtration rate (GFR), whereas in renal insufficiency group (with subtotal nephrectomy), following transient increases in ERPF and GFR, CM markedly decreased ERPF (-46.5 +/- 6.7%) and GFR (-51.2 +/- 7.1%). In sham operated group, theophylline and KF17837 markedly attenuated CM-induced increases in ERPF and GFR, while KW-3902 had no effects on CM-induced increases in ERPF or GFR. In renal insufficiency group, initial increases in ERPF and GFR were blunted by theophylline and KF17837. In contrast, the subsequent decreases in ERPF and GFR were attenuated by theophylline (% delta ERPF, -12.2 +/- 3.2% vs. -46.6 +/- 6.7%, P < 0.01; % delta GFR, 4.3 +/- 2.5% vs. -51.0 +/- 7.1%, P < 0.01), and were completely prevented by KW-3902 (% delta ERPF, 10.8 +/- 2.9%; % delta GFR, 23.8 +/- 4.4%), whereas KF17837 aggravated ERPF (-73.3 +/- 5.3%) and GFR (-78.4 +/- 5.3%). These data indicate that in normal renal function, iohexol elicits renal vasodilation by activating mainly the adenosine A2 receptors. In contrast, in impaired renal function, CM induces both A2 and A1 activation; the former is associated with the initial renal vasodilation, while the latter is responsible for the sustained aggravation of renal hemodynamics. PMID- 8731084 TI - Lupus autoantibodies interact directly with distinct glomerular and vascular cell surface antigens. AB - We have identified monoclonal anti-DNA antibodies derived from lupus prone MRL lpr/lpr mice that produce glomerular immune deposits and nephritis after passive transfer to normal mice. Particularly noteworthy is that the location of immune deposition varied among nephritogenic Ig, and this was associated with distinctive histologies and clinical disease profiles. Although their autoantigen binding properties differed, they were highly cross-reactive, in a manner similar to Ig deposited in glomeruli of lupus mice. This antigen binding profile was also typical of other previously described nephritogenic autoantibodies that bound directly to glomerular antigens to initiate immune deposit formation. In this study, we questioned whether ligation of different glomerular antigens by individual autoantibodies could contribute to the observed differences in the location of immune deposits. To examine this possibility, monoclonal anti-DNA antibodies (IgG2a) that produced glomerular immune deposits in different locations were evaluated. H221 produced mesangial, intracapillary (that is, intraluminal or within the capillary lumen) and subendothelial deposits associated with heavy proteinuria, whereas H147 produced mesangial, subendothelial and linear basement membrane deposits associated with proliferative glomerulonephritis. Initially, the capacity of H221 and H147 to bind directly to glomerular and vascular cell surfaces was evaluated. As demonstrated by FACS, H221 bound preferentially to mesangial cells whereas H147 bound preferentially to endothelial cells. To identify possible target cell surface antigens, Western blots, immunoprecipitation of surface labeled cells, and 2D gel electrophoresis were employed. H221 reacted with a 108 kDa protein on mesangial cells not identified by H147, whereas H147 reacted with a 45 kDa protein on endothelial cells not identified by H221. These results support the hypothesis that some nephritogenic lupus autoantibodies initiate immune deposit formation through direct interaction with glomerular antigens. Furthermore, they suggest that the site of immune deposition is determined by both antigen binding properties of the relevant antibody and the location of its target ligand within the glomerulus. In a given individual, therefore, the predominant autoantibody glomerular antigen interaction may influence the morphologic and clinical phenotype expressed. Variation in the predominant interaction may also contribute to variations in disease expression among individuals with lupus nephritis. PMID- 8731085 TI - Heterogeneous T cell receptor V beta gene repertoire in murine interstitial nephritis. AB - Anti-tubular basement membrane disease (alpha TBM) produces T cell-mediated interstitial nephritis in SJL/J mice following immunization with heterologous renal tubular antigen. Initial mononuclear infiltrates appear in vivo after six to eight weeks, with subsequent progression to renal fibrosis and endstage kidney disease. Cultured lymph node derived nephritogenic T cells from these mice react to a small epitopic region of the 3M-1 target antigen and share a common amino acid motif in their V beta CDR3 regions. We now have used RT-PCR to further characterize the renal expression of T cell receptor (TcR) V beta gene repertoires during the course of this disease. Individual kidneys with focal mononuclear infiltrates characteristic of early alpha TBM disease express up to three different TcR V beta genes; however, the same V beta genes are not found in all kidneys at the same early stage of injury. DNA sequencing of the V beta RT PCR products reveals a heterogeneous population of VDJ recombinations and deduced CDR3 amino acid sequences. Our studies do not support TcR V beta region gene restriction in histologically-detectable alpha TBM disease, but are more consistent with a dynamic, organ-specific autoimmune disease, directed at multiple autoantigenic epitopes. PMID- 8731086 TI - Expression of a nonmuscle myosin heavy chain in glomerular cells differentiates various types of glomerular disease in rats. AB - To characterize the phenotypic modulation of mesangial and glomerular epithelial cells, we investigated the expression of a nonmuscle type myosin heavy chain, SMemb, and alpha-smooth muscle actin (alpha-SM actin) in rat experimental glomerular diseases, which included anti-Thy 1 nephritis, 5/6 nephrectomy, diabetes, and anti-glomerular basement membrane nephritis. SMemb was only slightly expressed in normal glomerular epithelial cells but not in mesangial cells. In the anti-Thy 1 nephritis rats, both SMemb and alpha-SM actin were most conspicuously induced in mesangial cells. However, the expression profile was shifted from alpha-SM actin to SMemb dominant pattern over the course of glomerulonephritis. The expression of SMemb was also increased in epithelial cells in this model. In the other three models, glomerular cells did not express alpha-SM actin, but did so for SMemb. In the nephrectomized and the diabetic rats SMemb was newly expressed in mesangial cells at earlier stages, but at later stages was remarkably enhanced in epithelial cells when severe glomerular hypertrophy developed. In the anti-GBM nephritis rats, SMemb expression was increased in epithelial cells. In all models examined, mesangial and epithelial expression of SMemb was confirmed by immunoelectron microscopy, and enhanced expression of SMemb mRNA in glomeruli was verified by RNase protection assay. We conclude from these results that glomerular cells change their phenotypes differently depending on various types of glomerular diseases. These phenotypic changes in glomerular cells can be revealed by the combined immunostaining for SMemb and alpha-SM actin. SMemb is especially useful to detect both mesangial and glomerular epithelial cell activation in these glomerular disease models. Understanding the functional difference and regulatory mechanisms of these cytoskeletal proteins will provide insight into the pathogenesis and progression of glomerular diseases. PMID- 8731087 TI - Circadian rhythm in glomerular transport of macromolecules through large pores and shunt pathway. AB - In previous studies we have demonstrated that the circadian rhythm in renal clearance of serum proteins is more pronounced than the variability in glomerular filtration rate, and that the highest day-night fluctuations are found for the largest proteins. To analyze whether additional circadian rhythmicity in size selective glomerular transport could explain these phenomena, we measured renal clearances of inulin and dextrans in a range of 30 to 90 A over a period of one day and compared these data with renal clearance in proteins. Eight patients with nephrotic syndrome and a GFR > 60 ml/min and 6 healthy volunteers were studied during a protocol of bed rest and spaced protein and fluid intake. After administration of a loading dose, inulin and dextran were continuously infused. Blood and urine were sampled every three hours. In patients, but not in normals, fractional clearances of dextrans larger than 45 A showed a circadian rhythm with a peak in daytime and a close phase-relationship with the rhythm in GFR. The day night differences were the most pronounced for the largest dextrans. Analysis of the day-night differences in a computer model showed circadian variability in transport through the shunt pathway and through large pores. These results can, to an important degree, explain our previous observations on circadian variability in renal clearance of proteins in patients with nephrotic syndrome. PMID- 8731088 TI - Interaction of native and oxidized lipoprotein(a) with human mesangial cells and matrix. AB - The trapping of apolipoprotein(a) and apolipoprotein B-100 in glomeruli of patients with the nephrotic syndrome seems to be linked to a less favorable course of renal disease. To evaluate the potential role of lipoprotein(a) as a mediator of glomerular injury, we measured uptake of native lipoprotein(a) [Lp(a)] and oxidatively modified Lp(a) by cultured human mesangial cells and matrix and studied the effects of Lp(a) on mesangial cell DNA-synthesis and cellular proliferation. Uptake of Lp(a) by mesangial cells occurred at a significantly lower affinity (KD 16 micrograms/ml vs. 39 micrograms/ml) and a lower maximum degradative capacity (6.7-fold) than for LDL. Specificity of receptor mediated uptake was 50% for Lp(a) compared to 84% for LDL. Oxidative modification of both Lp(a) and LDL was accompanied by a significant decrease in uptake and degradative capacities. Due to the limited uptake, native and oxidatively modified Lp(a) had only marginal effects on intracellular cholesterol metabolism, which was measured as inhibition of sterol synthesis and stimulation of cholesterol esterification. However, binding of Lp(a), oxidized Lp(a) and oxidized LDL to extracellular mesangial matrix was enhanced compared to LDL. Furthermore, incubation of mesangial cells with Lp(a) and oxLp(a) in concentrations of 2.5 micrograms/ml and higher resulted in a decrease of DNA synthesis. Regardless of the oxidative status, a maximal suppression of DNA synthesis was observed at 20 micrograms/ml Lp(a). Native Lp(a) also blunted the stimulatory effects of PDGF on mesangial cell DNA-synthesis. Lp(a) did not alter basal TGF-beta transcription in human mesangial cells. The avid interaction of Lp(a) and modified lipoproteins with mesangial matrix provides a concept for the enhanced entrapment of these lipoproteins in the diseased glomerulum. Native Lp(a) is a poor ligand for the LDL receptor; oxidation of Lp(a) even lowers the affinity towards this receptor. Further studies must be carried out to clarify the pathophysiological significance of Lp(a) trapping in the mesangial matrix. PMID- 8731089 TI - Effect of sodium bicarbonate on intracellular pH under different buffering conditions. AB - Previous in vitro studies have reported a paradoxical exacerbation of intracellular acidosis following bicarbonate therapy due to the generated CO2 entering the cytoplasm. However, these studies were conducted in nonphysiological Hepes-buffered media. We compared the effect of a sodium bicarbonate load on the intracellular pH (pHi) of hepatocytes placed in nonbicarbonate (NBBS) or bicarbonate (BBS) buffering systems. The pHi of isolated rat hepatocytes was measured using the fluorescent pH sensitive dye BCECF and a single-cell imaging technique. Cells were placed in medium buffered with HCO3-/CO2 or Hepes. All media were adjusted to pH 7 with L-lactic acid or HCl. An acute 45 mM sodium bicarbonate load was added to each medium and the changes in pHi were measured every three seconds for 90 seconds. The sodium bicarbonate load caused rapid cytoplasmic acidification of cells in NBBS (N = 50, P < 0.001). In contrast, hepatocytes in BBS underwent a marked increase in pHi (N = 50, P < 0.001) without any initial decrease in pHi. These differences were highly significant for the buffer (P < 0.01), but not for the acid used. We conclude that sodium bicarbonate exacerbates intracellular acidosis only in a NBBS. Hence, in vitro studies reporting a paradoxical intracellular acidosis following bicarbonate therapy cannot be extrapolated to the in vivo buffering conditions, and should not be used to argue against bicarbonate therapy. PMID- 8731090 TI - Effects of contrast media and mannitol on renal medullary blood flow and red cell aggregation in the rat kidney. AB - Hemodynamic factors may play a role in the development of acute renal failure following administration of contrast media (CM). In this study the effect of intravenous injection of contrast media and mannitol on red blood cell velocity (VRBC) and red blood cell aggregation in renal medullary vessels was studied in 58 rats. Renal medullary blood flow was investigated by a cross-correlation technique and by a visual aggregation score. The CM, namely diatrizoate, iopromide, iohexol, ioxaglate, iotrolan, were given in iodine equivalent doses (1600 mg/kg body wt). Mannitol (950 mOsm/liter) and Ringer's solution were used as controls. The same vessels were studied 30 minutes before and 30 minutes after injections. VRBC decreased significantly after injection of diatrizoate, iopromide, iohexol, iotrolan and mannitol. Ringer's solution and ioxaglate did not significantly alter medullary blood flow, while iotrolan and mannitol caused the largest decreases in VRBC. All CM and mannitol caused both red cell aggregation and cessation of blood flow. The decrease in blood flow and increase in red blood cell aggregation after injection of CM and mannitol may partly explain the occurrence of contrast medium-induced acute renal failure. PMID- 8731091 TI - Influence of peritoneal dialysis solutions with glucose and amino acids on ingestive behavior in rats. AB - Continuous ambulatory peritoneal dialysis (CAPD) is often associated with malnutrition; reduced intake of nutrients due to anorexia is an important factor. The glucose load from glucose-based peritoneal dialysis (PD) solutions and amino acids from amino acid-based solutions may favor suppression of the appetite. To study this matter we used a new experimental model in free-moving, unstressed male Wistar rats (300 to 350 g) with feeding catheters channeled from the top of the skull to the oral cavity. When the rats recovered from surgery they were tested under standardized conditions by giving them an intraoral infusion (1 ml/min) of a solution containing 342 g/liter of the sucrose or 97 g/liter protein solutions while recording the time (volume) of ingestion. Control rats consumed 18.8 +/- 0.9 ml of the sucrose and 39.8 +/- 0.8 ml of the protein solutions. Injections of PD solutions with 13.6, 22.7, and 38.6 g/liter of glucose reduced the ingestion of sucrose by 12.4%, 23.6% and 36.1%, respectively, but did not affect the ingestion of protein. Injections of 30 ml of PD solutions containing 11, 18 and 31 g/liter of amino acids reduced the ingestion of both sucrose by 9.7%, 17.1% and 33.2% and of protein by 13.5%, 25.9% and 33.1%, respectively. We conclude that in our experimental model, the inhibition of appetite caused by peritoneal solutions containing glucose or amino acids seems to be specific for each nutritional constituent and not simply an effect of hyperosmolality or large filling volumes. PMID- 8731092 TI - Cyclosporine induces glomerulosclerosis: three-dimensional definition of the lesions in a rat model of renal transplant. AB - Previous description of chronic cyclosporine (CsA) renal toxicity commonly included vascular changes, tubular atrophy, and interstitial fibrosis. Glomerular injury was only occasionally documented and it is not clear whether glomerular changes may have an impact on the clinical syndrome of CsA toxicity observed in experimental animals and humans. At the moment the prevailing view is that when patients given CsA have glomerular lesions at a renal biopsy, these were due to concomitant chronic rejection or renal hypoperfusion rather than to CsA itself. As a follow-up of our previous work on the subject (abstract; J Am Soc Nephrol 2:1398, 1992) the present study was undertaken to clarify whether CsA is a direct cause of glomerular injury. We used a model of renal transplant among Lewis rats to better mimic the condition in which CsA is given to humans. Animals underwent kidney isografts and were given daily CsA or vehicle for as long as 12 months. At the end of the experiment specimens of renal tissue were analyzed by a serial section morphometric analysis technique, which allows precise evaluation at the individual glomerular level, glomerular volume and percentage of the capillary tuft affected by sclerosis. Among 85 glomeruli from CsA-treated rats, examined by three-dimensional morphometric analysis, only 12% were normal and 88% revealed segmental sclerosis. Data of single-section analysis compared with three dimensional morphometric reconstruction showed that the former markedly underestimated the extent of glomerular injury. By three-dimensional analysis we showed that chronic CsA administration was associated with profound changes in glomerular capillary tuft volume distribution as compared to normal. Specifically, a subset of smaller than normal glomeruli emerged in CsA-treated animals in addition to a population of glomeruli which were larger than normal. No significant correlation was found between capillary tuft volume and sclerosis volume. These findings indicate that chronic administration of CsA induces in rats glomerular lesions comparable to the ones reported in human renal or heart transplant. Our present model may help investigating the mechanism(s) of chronic CsA renal toxicity, and will provide important clues for pharmacological manipulations aimed at reducing the long-term consequences of CsA on the kidney. PMID- 8731093 TI - Hypoxia-induced amphiphiles inhibit renal Na+, K(+)-ATPase. AB - We have characterized the effects of hypoxia on carnitine metabolism in proximal tubules. Hypoxia for 10 minutes resulted in a significant increase in the mass of long chain acylcarnitines (LCAC) (control 53 +/- 20 vs. hypoxia 118 +/- 38 pmol. mg-1 protein). Since LCAC are proximal metabolites in the beta-oxidation of fatty acids, these data suggest that inhibition of fatty acid oxidation occurs during hypoxia in the proximal tubule. In addition to LCAC accumulation, hypoxia resulted in a significant increase in the mass of lysoplasmenylcholine LPLasCho (control 62 +/- 15 pmol/mg vs. 20 min hypoxia 146 +/- 21 pmol/mg protein, N = 4) and also in increases in the mass of monoacyl LPC (control 122 +/- 24 pmol/mg protein vs. 283 +/- 35 pmol/mg protein after 40 min of hypoxia). We tested the possibility that these compounds that accumulate during hypoxia could inhibit proximal tubule Na+, K(+)-ATPase. LPC, LPlasC, and LCAC inhibited proximal tubule nystatin-stimulated oxygen consumption (QO2) and proximal tubule Na+, K(+)-ATPase activity in a dose-dependent manner. In addition, LPC, LPlasC, and LCAC directly inhibited' (65%, 80%, and 60%, respectively) Na+, K(+)-ATPase activity purified from kidney cortex at similar concentrations at which they accumulate during hypoxia (above 25 microM). The present data suggest that amphiphile accumulation may have a potential pathophysiologic role in the proximal tubule during renal ischemia. PMID- 8731094 TI - Intragraft TGF-beta 1 mRNA: a correlate of interstitial fibrosis and chronic allograft nephropathy. AB - Chronic allograft nephropathy is a relentlessly progressive process and a major cause of long-term graft dysfunction and ultimate failure. Interstitial fibrosis, tubular atrophy, and glomerular and vascular lesions characterize this mechanistically unresolved disorder. Given the prominent role of TGF-beta 1 in tissue repair and in fibrosis, we have explored the hypothesis that fibrosis and chronic allograft nephropathy would be distinguished by intragraft TGF-beta 1 mRNA expression. This postulate was tested by mRNA phenotyping of RNA isolated from 127 human renal allograft biopsies. Reverse transcription assisted polymerase chain reaction was used to amplify and identify ingraft gene expression. Our investigation demonstrated a significant correlation between intragraft TGF-beta 1 mRNA display and renal allograft interstitial fibrosis and chronic allograft nephropathy. In contrast, intragraft expression of mRNA encoding immunoregulatory cytokines, IL-2, IFN-gamma, IL-4, IL-10, or cytotoxic attack molecules, granzyme B and perforin was not a correlate of interstitial fibrosis or chronic allograft nephropathy. Our studies identify, for the first time, a significant association between intragraft TGF-beta 1 mRNA expression and renal allograft interstitial fibrosis, and advance a candidate molecular mechanism for chronic allograft nephropathy. PMID- 8731095 TI - Advanced oxidation protein products as a novel marker of oxidative stress in uremia. AB - Evidence suggests an imbalance between antioxidant and oxidant-generating systems resulting in oxidative stress in uremic patients. As plasma proteins are critical targets for oxidants, we developed a novel spectrophotometric assay which allows to detect advanced oxidation protein products (AOPP) in uremic plasma. By size exclusion chromatography AOPP are retrieved in two distinct peaks at 600 and below 80 kDa in uremic plasma, while no such peaks are found in control plasma. Further biochemical characterization revealed that AOPP are carried by oxidized plasma proteins, especially albumin and do not have oxidant properties. AOPP increased in a dose-dependent manner following in vitro exposure of plasma or purified human serum albumin (HSA) to hypochlorous acid. Advanced glycation end products of human serum albumin (AGE-HSA) also increased AOPP levels. In vivo, plasma level of AOPP was the highest in patients on hemodialysis, followed by those on peritoneal dialysis and by undialyzed patients with advanced chronic renal failure. AOPP levels correlated with plasma concentrations of dityrosine and AGE-pentosidine, as indices of oxidant-mediated protein damage, but not with thiobarbituric reactive substances as lipid peroxidation markers. A close correlation was also found between AOPP and neopterin levels, suggesting that AOPP could be part in the monocyte-mediated inflammatory disorders associated with uremia. In conclusion, we propose the measurement of AOPP as a reliable marker to estimate the degree of oxidant-mediated protein damage in uremic patients and to predict the potential efficacy of therapeutic strategies aimed at reducing such an oxidative stress. PMID- 8731096 TI - Nitric oxide kinetics during hypoxia in proximal tubules: effects of acidosis and glycine. AB - In the present study, we directly monitored nitric oxide (NO) with an amperometric NO-sensor in suspensions of rat proximal tubules. Hypoxia-stimulated NO generation was characterized by an initial rise and a subsequent sustained increase which preceded cell membrane damage as assessed by lactic dehydrogenase (LDH) release. In contrast, the NO concentration remained unmeasurable in normoxic controls. Nitro-L-arginine-methyl ester (L-NAME) prevented the hypoxia induced increase in NO in a dose dependent manner in parallel with incremental cytoprotection. The hypoxia-induced elevation in NO and the associated membrane injury were both markedly prevented by extracellular acidosis (pH 6.95). In vitro proximal tubular nitric oxide synthase (NOS) activity (3H-arginine to 3H citrulline assay) was pH dependent with optimum activity at pH 8.0 and greatly reduced activity at acidic pH even in the presence of calcium and co-factors. However, glycine, a well recognized cytoprotective agent, did not attenuate the NO concentration during hypoxia. The present study therefore provides direct evidence that NO is generated by rat proximal tubules during hypoxia and demonstrates that the protective effect of low pH against hypoxic rat tubular injury is associated with an inhibition of this NO production. PMID- 8731098 TI - Differentiation of smooth muscle phenotypes in mouse mesangial cells. AB - Smooth muscle alpha-actin (SMA) mRNA, a marker of vascular smooth muscle cells, was identified in the normal glomerular mesangium both in vivo and in vitro. Several populations of mesangial cells were studied to determine if SMA and basement membrane collagen were regulated together. The levels of SMA expression, which could be linked to the stage of differentiation, were different for the differing cell populations. One cell population had high SMA and type IV collagen levels at its early passages. The others expressed both interstitial and basement membrane collagens. The first population developed these phenotypic features at later passages. The levels of SMA and alpha 1(IV) collagen expression were regulated together in concert, whereas the alpha 2(I) collagen levels were expressed inversely to SMA and alpha 1(IV) collagen. Both SMA and type IV collagen were controlled by the methylation states of the cis-regulators; however, type I collagen was mainly regulated by the trans-acting regulators. Treatment with 5-azacytidine converted the cells of a fibroblast-phenotype to a smooth muscle cell-like phenotype. These cell lines may be useful for studying the differentiation process in vitro. PMID- 8731097 TI - Role of endothelin as a mitogen in experimental glomerulonephritis in rats. AB - Recent studies have revealed that endothelin-1 (ET-1) is a potent mitogen for mesangial cells in vitro. To determine whether ET-1 exerts the mitogenic action on mesangial cells in vivo, we examined the glomerular expression of ET-1 and its receptors in a rat model of mesangial proliferative glomerulonephritis and assessed the effect of a specific endothelin A (ET(A)) receptor antagonist, FR139317, on mesangial cell proliferation in this model. The levels of preproET-1 mRNA expression and ET-1 protein production in glomeruli increased markedly on days 4 and 7 after disease induction, and the levels changed in concordance with the glomerular cell proliferation. In contrast, the level of ET(A) receptor mRNA initially decreased on day 1, and thereafter increased on days 4 and 7. Administration of FR139317 to rats with experimental glomerulonephritis induced a significant reduction in mesangial cell proliferation. In addition, in situ hybridization of preproET-1 mRNA and double-immunolabeling of ED-1 and OX-7 in a mirror image section revealed that the principal cell expressing ET-1 in glomeruli were infiltrating macrophages on day 1, and they were replaced by mesangial cells on day 4. These findings indicate that ET-1 functions as a potent mitogen for mesangial cells in vivo in an autocrine or paracrine fashion. PMID- 8731099 TI - Acute passive anti-glomerular basement membrane nephritis in P-selectin-deficient mice. AB - P-selectin present on surfaces of activated endothelium and platelets mediates neutrophil-endothelial and neutrophilplatelet interactions. The role of P selectin in vivo was examined in a model of acute passive anti-GBM nephritis in P selectin-deficient and wild-type mice which was induced by intravenous injection of anti-GBM serum. There were two major differences between P-selectin-deficient and wild-type mice. Firstly, mutant mice had approximately two fold more glomerular PMNs and albuminuria than wild-type animals at the peak of neutrophil influx and proteinuria. Secondly, Lipoxin A4 (LXA4), an eicosanoid which inhibits leukocyte-endothelial adhesion in vitro, and is generated primarily by transcellular biosynthetic routes during P-selectin-mediated platelet-PMN interaction [1], was approximately 60% of wild type levels in nephritic kidneys of P-selectin-deficient mice. Injection of wild-type platelets into P-selectin null mice restored LXA4 to wild-type levels. The corresponding PMN influx approximated PMN levels in wild-type mice receiving platelets but urine albuminuria remained higher. Although these two P-selectin-dependent events cannot be directly linked, our results point to the importance of considering both platelet and endothelial P-selectin in determining the cellular events in inflammation. PMID- 8731100 TI - Increased expression of a brain/embryo-type myosin heavy chain isoform (MIIB2) in mesangial proliferative glomerulonephritis. AB - Proliferation of mesangial cells is frequently found in glomerulonephritis, such as IgA glomerulonephritis. Recent reports suggest that a brain/embryo-type myosin isoform (MIIB2) is involved in cell proliferation. We have studied the expression of MIIB2 in renal biopsy samples from patients with various renal diseases and in the renal tissues from the rat model of mesangial proliferative glomerulonephritis induced with anti-Thy 1.1 antibody. Immunohistochemical analysis of the biopsy samples using an anti-brain-type myosin heavy chain specific monoclonal antibody (HBM1) indicated that 92% of the samples from patients with IgA glomerulonephritis contained a significant population of mesangial cells that reacted with the antibody. Most of the samples from patients with other types of proliferative glomerular diseases also contained HBM1 reactive mesangial cells, while none of the samples from patients with non proliferative glomerular diseases contained a significant population of HBM1 reactive mesangial cells. The expression of a brain/embryo-type myosin heavy chain isoform (MIIB2) in the mesangial cells began at five days after injection of anti-Thy 1.1 antibody and peaked at the tenth day. On the other hand, the expression of the proliferating cell nuclear antigen in the mesangial cells was induced at two days after injection of anti-Thy 1.1 antibody and was maximal at the fourth day. These results indicate that the expression of the MIIB2 isoform by mesangial cells is accelerated in proliferative glomerulonephritis and suggest that the myosin isoform is involved in the phenotypic transformations of the glomerular tissues rather than in the cell proliferation. PMID- 8731101 TI - Increased lipase inhibition in uremia: identification of pre-beta-HDL as a major inhibitor in normal and uremic plasma. AB - The hypertriglyceridemia commonly observed in uremia has been attributed to an abnormally high inhibitor activity in plasma for lipoprotein lipase (LPL) and hepatic lipase (HL), both of which have a key role in lipoprotein metabolism. The purpose of this investigation was to establish a relationship between plasma lipase inhibitor activity and hypertriglyceridemia, identify the main plasma lipase inhibitor, and determine the basis for the greater inhibitor activity in uremia. In a mixed population of normal (N = 8) and uremic subjects (N = 12), log transformed plasma triglycerides correlated with both inhibitor activity and uremic status. However, inhibitor activity was the only retained predictor variable for triglycerides in a multiple linear regression model (r = 0.91; P < 0.0001). An inhibitor isolated from normal plasma was identified as a particle containing apolipoprotein A-I (apo A-I) and 3% phospholipid. This particle, which has pre-beta electrophoretic mobility and a Stokes' radius of 54 A, therefore corresponds to a form of the previously described pre-beta-HDL (free apo A-I) in the non-lipoprotein fraction of plasma. Comparison of normal and uremic plasma indicated that the greater lipase inhibitor activity in the latter could be attributed to an increased concentration of apo A-I in the non-lipoprotein fraction of plasma (pre-beta-HDL), as well as to increased inhibition by the uremic lipoproteins. The increased plasma lipase inhibitor activity may be important in the pathogenesis of hypertriglyceridemia in chronic renal failure. PMID- 8731102 TI - Studies on the mechanism of trimethoprim-induced hyperkalemia. AB - We examined the effects of trimethoprim (TMP) on metabolic parameters and renal ATPases in rats after a 90 minute infusion (9.6 mg/hr/kg body wt, i.v.) and after 14 days (20 mg/kg body wt/day, i.p.). After one dose of TMP, plasma electrolytes, arterial pH and aldosterone levels were normal, but a natriuresis, bicarbonaturia, and decreased urinary potassium excretion occurred. Na-K-ATPase activity in microdissected segments from these animals was decreased by 36 +/- 0.9% in proximal convoluted tubule (PCT) (P < 0.005); decreases of 50 +/- 2.1% and 40 +/- 1.1% were seen in cortical and medullary collecting tubules (CCT and MCT), respectively (P < 0.005). Na-K-ATPase activity was unaffected in medullary thick ascending limb (MTAL). H-ATPase (in PCT and collecting duct) and H-K-ATPase (in CCT and MCT)-activities were not changed. Following chronic TMP administration, plasma potassium increased as compared to control (5.16 +/- 0.05 mEq/liter vs. 3.97 +/- 0.05 mEq/liter, P < 0.05), however, acid-base status and plasma aldosterone levels were normal. Na-K-ATPase activity was decreased by 45 +/- 2.6% in PCT (P < 0.005), 73 +/- 2.0% in CCT (P < 0.001), and 53 +/- 2.5% in MCT (P < 0.005). Na-K-ATPase, activity in MTAL and H-K-ATPase activity in CCT and MCT were unchanged. H-ATPase activity in PCT and MTAL was normal, but in the collecting tubule (CCT and MCT) it was decreased by approximately 25% (P < 0.05). TMP inhibited Na-K-ATPase activity in a dose-dependent fashion in PCT, CCT, and MCT when tubules from normal animals were incubated in vitro with the drug; TMP in vitro did not affect H-ATPase or H-K-ATPase activity. These results suggest that TMP-induced hyperkalemia may result from decreased urinary potassium excretion caused by inhibition of distal Na-K-ATPase, in the face of intact H-K ATPase activity. PMID- 8731103 TI - Impact of hypertension on cardiomyopathy, morbidity and mortality in end-stage renal disease. AB - A cohort of 432 ESRD (261 hemodialysis and 171 peritoneal dialysis) patients was followed prospectively for an average of 41 months. Baseline and annual demographic, clinical and echocardiographic assessments were performed, as well as serial clinical and laboratory tests measured monthly while on dialysis therapy. The average mean arterial blood pressure level during dialysis therapy was 101 +/- 11 mm Hg. After adjusting for age, diabetes and ischemic heart disease, as well as hemoglobin and serum albumin levels measured serially, each 10 mm Hg rise in mean arterial blood pressure was independently associated with: the presence of concentric LV hypertrophy (OR 1.48, P = 0.02), the change in LV mass index (beta = 5.4 g/m2, P = 0.027) and cavity volume (beta = 4.3 ml/m2, P = 0.048) on follow-up echocardiography, the development of de novo cardiac failure (RR 1.44, P = 0.007), and the development of de novo ischemic heart disease (RR 1.39, P = 0.05). The association with LV dilation was of borderline statistical significance (OR 1.48, P = 0.06). Mean arterial blood pressures greater than 106 mm Hg were associated with both echocardiographic and clinical endpoints. Paradoxically, low mean arterial blood pressure (RR 1.36 per 10 mm Hg fall, P = 0.009) was independently associated with mortality. The association of low blood pressure with mortality was a marker for having had cardiac failure prior to death. We conclude that even moderate hypertension worsens the echocardiographic and clinical outcome in ESRD patients, especially in those without previous clinical cardiac disease. PMID- 8731104 TI - Continuous ambulatory peritoneal dialysis impairs T lymphocyte selection in the peritoneum. AB - Peritoneal lymphocytes (PCL) of 45 healthy individuals, four uremic patients with end-stage renal disease (ESRD) and 25 long-term continuous ambulatory peritoneal dialysis (CAPD) patients were characterized by flow cytometry to investigate whether CAPD alters the phenotype of PCL. B lineage cells constitute a minority of PCL (2.5% of cells). Although the majority of peritoneal T cells expressed alpha beta T cell receptor (TcR), 7% expressed gamma delta TcR, a proportion which was significantly higher than that in peripheral blood (PBMC) (approximately 4%). The majority of PCL T cells exhibited markers of the thymus dependent lineage (CD2, CD3, TcR alpha beta, CD8 alpha beta or CD4) and surface antigens associated with memory and activation (CD45RO, CD11a, CD18, CD49d, HLA DR). An average of 75% of both CD4+ and CD8+ PCL T cells of healthy subjects and CAPD patients were CDw60+, thus characterizing the T cell subset containing the helper activity for the mitogen-driven B cell differentiation. CD44s was abundantly expressed on PCL T cells. In contrast to PCL T cells of healthy subjects peritoneal T lymphocytes of CAPD patients exhibited CD44 splice variants containing products of exon-v9 and the proportion of CD44v9+ cells correlated with the frequency of peritonitis episodes the patients had gone through. The majority of PCL T cells of both healthy subjects and CAPD patients were CD8+. A large proportion of CD8+ PCL T cells from healthy subjects expressed the homodimeric CD8 alpha alpha isoform; however, such cells were not found in CAPD patients. In healthy subjects mRNA for the recombination activating gene 1 (RAG 1) was detectable in a PCL population containing CD7-CD34+ and CD7+CD34+ cells. In contrast, neither mRNA transcripts of the RAG-1 gene nor CD34+ cells were detectable in PCL of CAPD patients. PMID- 8731105 TI - Correction of acidosis in CAPD decreases whole body protein degradation. AB - Correction of acidosis in CAPD decreases protein degradation and synthesis but has no effect on leucine oxidation. The effect of the correction of metabolic acidosis in CRF patients treated with CAPD was determined from the kinetics of infused L-[1-13C]leucine. Seven CAPD patients were studied before (acid) and after correction of acidosis (bicarbonate) (pH:acid 7.39 +/- 0.01, bicarbonate 7.41 +/- 0.01, P = 0.005). Leucine appearance from body protein (PD) [corrected] and leucine disappearance into body protein (PS) [corrected] decreased significantly with correction of acidosis. (PS: acid 211.7 +/- 9.8, bicarbonate 142.3 +/- 4.2 micromol x kg-1 x hr-1, P < 0.001; PD: acid 200.6 +/- 8.5, bicarbonate 132.4 +/- 3.7 micromol x kg-1 x hr-1, P < 0.001). There was no significant change in leucine oxidation or plasma amino acid concentrations. These results demonstrate that optimal correction of acidosis in CAPD is beneficial in terms of protein turnover. PMID- 8731106 TI - Molecular genetics of cystinuria in French Canadians: identification of four novel mutations in type I patients. AB - Cystinuria, a hereditary disorder of cystine and dibasic amino acid reabsorption, has been classified into three subtypes on the basis of urinary excretion in obligate heterozygous parents. Thirteen cystinuric patients, identified primarily through the Quebec newborn urinary screening program, were investigated by phenotypic classification and by mutational analysis of the D2H (rBAT) gene. Mutations were identified on 7 of 25 alleles; all of these 7 mutant alleles were associated with Type I cystinuria. Four of the mutations (a large deletion, a 5'splice site mutation, a 2 bp deletion, and a nonsense mutation) have not been previously reported. These findings suggest that abnormalities in the D2H gene may account for only one subtype (Type I) of cystinuria, and that this subtype can be caused by a wide variety of population-specific mutations. PMID- 8731107 TI - Zinc modulates mononuclear cellular calcitriol metabolism in peritoneal dialysis patients. AB - Zinc has long been known to play a role in maintaining immunologic function. Hypozincemia, however, is common in patients with end-stage renal disease (ESRD) treated with continuous ambulatory peritoneal dialysis (CAPD). We previously demonstrated that zinc depletion limits the ability of animals to achieve maximum circulating calcitriol levels in response to the stress of calcium or phosphorus depletion. It was unclear, however, whether changes in the circulating levels of calcitriol in these settings was associated with a direct effect on renal 1-alpha hydroxylase activity, or whether the zinc dependence of the stimulated calcitriol response involved an integrated systemic response in intact animals. In addition it was unclear whether circulating zinc levels or zinc nutritional status modified calcitriol metabolism in humans. To better understand the role zinc plays in the immune response in patients with ESRD, we studied IL-1, calcitriol and tumor necrosis factor-alpha production by mononuclear cells from blood and peritoneal effluents of 22 patients with ESRD treated with CAPD. Macrophages from peritoneal effluents and peripheral blood mononuclear cells were isolated and pulsed with phytohemagglutinin in medium to which different concentrations of zinc chloride, copper chloride, and carbonyl cyanide p-(trifluoromethoxy)-phenyl hydrazone (FCCP), an inhibitor of mitochondrial function were added. Supernatant interleukin-1, calcitriol, and tumor necrosis factor-alpha levels were subsequently measured. We demonstrated a zinc concentration dependent increase in stimulated IL-1 alpha and -beta, and TNF-alpha release in both peripheral mononuclear cells and peritoneal macrophages from patients with ESRD treated with CAPD. The effect is zinc specific, as it is not reproduced by copper or chloride supplementation. A zinc concentration dependent increase in peritoneal macrophage calcitriol release was also noted. FCCP blocked the cellular production of IL-1 alpha, IL-1 beta, and TNF-alpha, but had little effect on zinc-induced stimulated mononuclear cell supernatant calcitriol levels. The different shape of the zinc dose response curve, and the lack of correlation between paired IL-1 and calcitriol supernatant levels suggests the effect of zinc on mononuclear cellular cytokine and calcitriol production is mediated through different pathways. PMID- 8731108 TI - Growth promoting effects of growth hormone and IGF-I are additive in experimental uremia. AB - Exogenous growth hormone (GH) stimulates the endogenous production of IGF-I and improves growth in uremia. We investigated whether exogenous IGF-I is also able to improve uremic growth failure in rats and whether the growth promoting effects of GH and IGF-I are additive. In female 150 g uremic (subtotal nephrectomy, NX) Sprague-Dawley rats, both rhGH in doses from 2 X 1.25 to 2 X 10 IU/kg bid s.c. and rhIGF-I in doses from 2 X 0.5 to 2 X 4.0 mg/kg bid s.c. caused a dose dependent increase in weight gain and length gain. However, endogenous production of GH was suppressed by both agents. Peptide hormone treatment did not affect cumulative food intake, but significantly increased food efficiency ratio (weight gain/food intake). Concomitant s.c. treatment with maximally effective doses of rhGH (12 X 5 IU/kg bid) and of rhIGF-I (2 X 2 mg/kg bid) resulted in additive growth promoting effects in NX and pair-fed control (CO) animals during the observation period of 12 days. Cumulative length gain was 3.2 +/- 0.5 cm in solvent-treated NX-animals, 4.1 +/- 0.5 cm with rhGH (+ 28% above solvent), 4.2 +/- 0.6 cm with rhIGF-I (+ 31%) and 4.9 +/- 0.5 cm with both peptides (+ 53%). The food efficiency ratio was 0.16 +/- 0.05 in solvent NX, 0.33 +/- 0.04 with rhGH (+ 106% above solvent), 0.23 +/- 0.02 with rhIGF-I (+ 44%), and 0.38 +/- 0.02 with both peptides (+ 138%). Histomorphometric analysis and measurements of length gain by fluorescence microscopy in the upper tibial metaphysis confirmed the growth promoting effects of both peptide hormones. The serum concentrations of IGF binding protein (BP)-4 (Western ligand blotting analysis) and of IGFBP-2 (immunoblot) were increased in uremic animals whereas IGFBP-3 was unchanged. Treatment with IGF-I and/or rhGH increased serum concentration of IGF-I but did not change the IGFBP pattern. rhIGF-I lowered blood glucose levels within one to two hours after injection. The effect was most pronounced during the first treatment day and declined thereafter. Concomitant treatment with rhGH attenuated the glucose lowering effect of rhIGF-I (glucose serum concentration at day one: 120 +/- 11 mg% in solvent NX, 50 +/- 21 mg% with rhIGF-I, 80 +/- 24 mg% with both peptides). It is concluded that: (i) IGF-I is able to stimulate growth in NX animals but suppresses endogenous GH production in the long run; (ii) the concomitant treatment with IGF-I and GH has additive effects on growth; and (iii) concomitant treatment with rhGH prevents hypoglycemia that is noted with rhIGF-I alone. PMID- 8731109 TI - Urinary kallikrein: a marker of blood pressure sensitivity to salt. AB - We evaluated if a rat strain inbred for low urinary kallikrein excretion differs from normal-kallikrein Wistar rats regarding blood pressure levels in basal conditions and during alterations in sodium balance. Blood pressure was measured in unanesthetized rats on normal sodium intake. Then, blood pressure sensitivity to salt was evaluated over a period of 20 days of high sodium diet (0.84 mmol per g chow). Low-kallikrein rats showed greater systolic blood pressure levels (125 +/- 3 vs. 114 +/- 2 mm Hg in controls, P < 0.01) at nine weeks of age. Systolic blood pressure was increased after sodium loading in the low-kallikrein group and remained unchanged in controls (150 +/- 6 vs. 112 +/- 2 mm Hg, P < 0.01). This effect was associated with a reduced cumulative urinary excretion of sodium in the low-kallikrein rats. No group difference was found in the clearance of endogenous creatinine in basal conditions. Urinary creatinine excretion decreased during sodium loading, particularly in the low-kallikrein group. The group difference in urinary kallikrein excretion found in basal conditions (6.85 +/- 0.31 vs. 20.74 +/- 1.71 nkat/24 hr in controls, P < 0.01) was enhanced by high salt diet (2.96 +/- 0.67 vs. 22.07 +/- 2.47 nkat/24 hr in controls, P < 0.01). In addition, renal kallikrein activity and content were reduced in low-kallikrein rats. The latter group showed a greater ratio of heart weight to body wt both in basal conditions and after sodium loading. The ratio of kidney weight to body wt was reduced after sodium loading. These results indicate that a genetically determined defect in urinary kallikrein excretion is associated with a greater blood pressure sensitivity to salt, possibly due to altered renal sodium handling. PMID- 8731110 TI - Outcome and risk factors of ischemic heart disease in chronic uremia. AB - To determine the prognosis and risk factors for ischemic heart disease in chronic uremia, a cohort of 432 dialysis patients were followed prospectively from start of dialysis therapy until death or renal transplantation. Baseline demographic, clinical and echocardiographic data were obtained. After the initiation of dialysis laboratory data were collected at monthly intervals, and clinical and echocardiographic data at yearly intervals. Twenty-two percent of patients (N = 95) had either a history of angina pectoris or myocardial infarction on starting dialysis therapy. Median time to onset of heart failure was 24 months in those with ischemic heart disease on initiation of dialysis, compared to 55 months in those without (P < 0.0001). This effect was independent of age, diabetes and underlying cardiomyopathy. Median survival was 44 months in those with ischemic disease compared to 56 months in those without (P = 0.0001). This adverse impact was independent of age and diabetes mellitus but, when cardiac failure was added to the Cox's model, ischemic heart disease was no longer an independent predictor of survival. De novo ischemic heart disease, not evident on starting dialysis therapy, occurred in 41 (9%) patients. When compared to patients who never developed ischemic disease (N = 296; 69%), significant and independent predictors of de novo disease were older age (P = 0.0007), diabetes mellitus (P = 0.0001), high blood pressure during follow up on dialysis (P = 0.02) and hypoalbuminemia (P = 0.03), whereas anemia was not an independent predictor. LV mass index was 174 +/- 7 g/m2 in those who developed de novo ischemic disease compared to 155 +/ 3 g/m2 (P < 0.001) in those who did not. Concentric LV hypertrophy, LV dilation and systolic dysfunction were independent risk factors for de novo ischemic heart disease. We conclude that ischemic heart disease occurs frequently in dialysis patients, that its adverse impact is mediated through the development of heart failure, and that the most important, potentially reversible risk factors are hypertension, hypoalbuminemia, and underlying cardiomyopathy. PMID- 8731111 TI - Neurocognitive function in chronic hemodialysis patients. AB - We administered a comprehensive neuropsychological test battery, including measures of intelligence, immediate and delayed memory, attention and mental processing speed, language abilities, complex problem solving, motor skills, and depression, to 16 well-dialyzed (Kt/Vurea = 1.46 +/- 0.24) patients with end stage renal disease (ESRD) and 12 age- and education-matched medical controls. We observed no clear neuropsychological deficits in these ESRD patients who had low average intelligence and limited educational achievement, and hypothesize that previously observed apparent deficits resulted either from very low dialysis delivery or comparison with poorly-matched historical controls. There were significant deficits in language ability and intelligence in ESRD patients with higher than median scores on the Beck Depression Inventory compared with less depressed ESRD patients. However, this effect of depression did not result in differences between dialysis and non-ESRD patient groups. PMID- 8731112 TI - Relative effects of PTH and dietary phosphorus on calcitriol production in normal and azotemic rats. AB - In moderate renal failure, the serum calcitriol level is influenced by the stimulatory effect of high PTH and the inhibitory action of phosphorus retention. Our goal was to evaluate the relative effect that high PTH levels and increased dietary phosphorus had on calcitriol production in normal rats (N) and rats with moderate renal failure (Nx). Normal and Nx (3/4 nephrectomy) rats were divided into two groups: (1) rats with intact parathyroid glands (IPTG) and (2) parathyroidectomized rats in which PTH was replaced (PTHR) by the continuous infusion of rat 1-34 PTH, 0.022 microgram/hr/100 g body wt, using a miniosmotic Alzet pump. To test the effect of dietary phosphorus, rats received either a moderate (MPD, 0.6% P) or a high phosphorus (HPD, 1.2%) diet for 14 days. The experimental design included pair-fed N and Nx rats with either IPTG or PTHR. Serum calcitriol and PTH levels in N rats fed a MPD were 69 +/- 3 and 40 +/- 5 pg/ml, respectively. In Nx rats on a MPD, serum calcitriol levels decreased only if hyperparathyroidism was not allowed to occur (76 +/- 4 vs. 62 +/- 4 pg/ml in Nx-IPTG-MPD and Nx-PTHR-MPD groups respectively, P < 0.05). Even in N rats on a HPD, high PTH levels (67 +/- 8 pg/ml in the N-IPTG-HPD group) were required to maintain normal serum calcitriol levels (69 +/- 4 vs. 56 +/- 6 pg/ml in Nx-IPTG HPD and Nx-PTHR-HPD groups, respectively; P < 0.05). In Nx rats on a HPD, the development of secondary hyperparathyroidism (286 +/- 19 pg/ml in the Nx-IPTG-HPD group) prevented a decrease in serum calcitriol levels (68 +/- 7 pg/ml). In contrast, serum calcitriol levels were low in the Nx-PTHR-HPD group (52 +/- 4 pg/ml, P < 0.05), which were deprived of the adaptative increase in endogenous PTH production. In conclusion, our results in rats indicate that in moderate renal failure, an elevated PTH level maintains calcitriol production and overcomes the inhibitory action of phosphorus retention. PMID- 8731113 TI - Biocompatibility of bicarbonate buffered peritoneal dialysis fluids: influence on mesothelial cell and neutrophil function. AB - The present study compares the effects of lactate and bicarbonate buffered PDF on human neutrophil (PMN) and human peritoneal mesothelial cell (HPMC) viability and function. Acute exposure of PMN to lactate buffered PDF at pH 5.5 (CAPD 2, 1.5% and CAPD 3, 4.25% glucose) resulted in significant reductions in cellular ATP levels, the phagocytosis of serum treated zymosan (STZ) and respiratory burst activation (CL). Exposure of PMN to bicarbonate buffered PDF (BIC 20, 1.5% glucose and BIC 30, 4.25% glucose both at pH 7.2) had no significant effect on cell viability or the CL response. Phagocytosis was, however, depressed significantly more following exposure to BIC 30 than BIC 20. PMN cellular ATP levels and phagocytosis were significantly better in cells exposed to BIC 30 than to CAPD 3 at pH 7.4 (P = 0.043 for both). Pre-exposure of HPMC to CAPD 2, CAPD 3 or BIC 30 for 30 minutes resulted in a significant reduction in cellular ATP content compared to control medium. Pre-exposure to BIC 20 did not result in a reduction in HPMC ATP levels. HPMC synthesis of IL-6 was unaffected by 15 or 30 minutes pre-exposure to BIC 20 or BIC 30, in contrast pre-exposure to CAPD 2 or CAPD 3 for 15 or 30 minutes resulted in a significant reduction in stimulated IL 6 synthesis (24.5 +/- 3.01 and 32.3 +/- 5.0 vs. 43.9 +/- 10 pg/microgram cell protein in M199, N = 6; P = 0.02). Neutralization of the pH of CAPD 2 and CAPD 3 resulted in normalization of HPMC IL-6 secretion. Analysis of IL-6 mRNA expression in control, BIC 20 and 30 pre-treated HPMC subsequently stimulated with IL-1 beta revealed no differences in the expression of the IL-6 specific 465 base pair transcripts. The improved cellular function in bicarbonate buffered PDF indicates potentially improved host defence status and preservation of the peritoneal membrane in CAPD patients. PMID- 8731114 TI - Sustained systemic arterial hypertension induced by extended hypobaric hypoxia. AB - Regular administration of recombinant erythropoietin (EPO) is frequently complicated by a rise in arterial blood pressure. We therefore asked if prolonged stimulation of endogenous EPO production has the same effect. To this end, male Sprague-Dawley rats were placed in a hypobaric chamber (390 mm Hg) for 24 days. The control (NL) group was placed in the chamber at normobaric condition. The animals were then removed from the chamber and monitored through day 108. Plasma EPO peaked within 24 hours and returned to baseline by day 7 and remained so thereafter. Hematocrit rose steadily during the hypoxic phase and declined steadily during the normobaric phase, reaching the baseline on day 45. This was accompanied by parallel changes in erythrocyte mass and blood volume. The rise in hematocrit during hypoxia was accompanied by a parallel rise in blood pressure which peaked on day 24. Despite the restoration of normal hematocrit, erythrocyte mass and blood volume following resumption of normoxia, blood pressure remained elevated throughout the observation period. To dissect the role of hypoxia from that of the associated rise in hematocrit, the experiments were repeated using a group of rats whose hematocrits were kept constant by repeated phlebotomies. These animals exhibited a sustained rise in blood pressure identical to that found in the original group. Thus, prolonged hypobaric hypoxia leads to a severe hematocrit-independent systemic hypertension (HTN) that persists long after the restoration of normoxia. Given the transient nature of the rise in its plasma concentration, endogenous EPO does not appear to play a role in the genesis of the observed systemic HTN. The authors believe that this animal model can be used for future studies of the mechanism, consequences and treatment of acquired HTN. PMID- 8731115 TI - Comparison of mortality with home hemodialysis and center hemodialysis: a national study. AB - We sought to determine whether lower mortality rates reported with hemodialysis (HD) at home compared to hemodialysis in dialysis centers (center HD) could be explained by patient selection. Data are from the United States Renal Data System (USRDS) Special Study Of Case Mix Severity, a random national sample of 4,892 patients who started renal replacement therapy in 1986 to 1987. Intent-to-treat analyses compared mortality between home HD (N = 70) and center HD patients (N = 3,102) using the Cox proportional hazards model. Home HD patients were younger and had a lower frequency of comorbid conditions. The unadjusted relative risk (RR) of death for home HD patients compared to center HD was 0.37 (P < 0.001). The RR adjusted for age, sex, race and diabetes, was 44% lower in home HD patients (RR = 0.56, P = 0.02). When additionally adjusted for comorbid conditions, this RR increased marginally (RR = 0.58, P = 0.03). A different analysis using national USRDS data from 1986/7 and without comorbid adjustment showed patients with training for self care hemodialysis at home or in a center (N = 418) had a lower mortality risk (RR = 0.78, P = 0.001) than center HD patients (N = 43,122). Statistical adjustment for comorbid conditions in addition to age, sex, race, and diabetes explains only a small amount of the lower mortality with home HD. PMID- 8731116 TI - Blood volume, colloid osmotic pressure and F-cell ratio in children with the nephrotic syndrome. PMID- 8731117 TI - Metanephros organogenesis is highly stimulated by vitamin A derivatives in organ culture. AB - Vitamin A and its metabolic derivatives are known to be key signalling molecules in regulating morphogenetic events in vertebrate development. Here we investigated their possible involvement during mammalian kidney development using paired rat metanephros organ culture. Metanephroi were explanted from 14-day-old embryos and cultured for six days in a chemically defined medium containing a retinoid at a dose of 10(-11) to 10(-4) M. Retinol, all-trans and 9-cis retinoic acid were able to promote metanephros growth and differentiation in vitro. A significant increase in the number of nephrons was observed from 10(-8) M of retinol and 10(-10) M of all-trans retinoic acid, before any change in growth parameters. A threefold increase in the number of nephrons was obtained at a dose of 10(-6) M. At low retinoid concentrations, there was a modulating effect of triiodothyronine on retinoid-stimulated nephrogenesis since the absence of triiodothyronine in the medium enhanced the nephrogenic stimulation. Exposure of metanephroi from 13-day-old embryos to all-trans retinoic acid (10(-7) M) led to a sixfold increase of nephron formation. Finally, we analyzed the branching pattern of the ureteric bud and showed that within 48 hours of culture, it was significantly more developed upon retinoid exposure. In conclusion, this study demonstrates that retinoic acid is a key regulator of renal organogenesis in controlling nephrogenic induction processes and ureteric bud patterning, and that the younger the metanephros, the greater the effect. PMID- 8731118 TI - Frederick Akbar Mahomed and his role in the description of hypertension at Guy's Hospital. PMID- 8731119 TI - New insights into the pathogenesis of idiopathic hypercalciuria: the role of bone. PMID- 8731120 TI - The sigma rectum pouch (Mainz pouch II). AB - A low-pressure reservoir for urine is created by antimesenteric splitting and side-to-side anastomosis of the rectosigmoid, the expectation being to obtain better continence rates and better protection of the upper tract than are achievable by ureterosigmoidostomy. Between 1990 and August 1993 the procedure was performed in 73 patients (59 adults and 14 children) whose mean age was 43.5 years. The indications were malignancy (n = 55), bladder exstrophy/epispadias (n = 14), trauma (n = 3), and sinus urogenitalis (n = 1). Of the 73 patients, 69 were followed for a mean period of 127 (range, 1-34) months. In all, 5 early complications were encountered (6.8%). In addition, 8 late complications occurred (10.9%), stenosis at the ureteral implantation site being the most common one. Daytime continence was 94.5% and night-time continence, 98.6%. The sigma rectum pouch achieves excellent continence rates. Despite implantation of the ureters into a low-pressure reservoir, stenosis at the site of ureteral implantation occurred in 6.8% of the patients, demonstrating the profund vulnerability of ureterointestinal anastomosis. PMID- 8731121 TI - The modified rectal bladder in children: long-term follow-up. AB - The criteria for evaluation of urinary diversion procedures in children must be strict since their life expectancy is long. Our experience with the modified rectal bladder in children with considerable follow-up periods is reported herein. All patients were continent by day and night. Urography studies revealed a normal upper tract in all cases. Three early complications were encountered among patients who had a submucous tunnel reimplantation. Reflux to the proximal colon or the kidneys was not demonstrated. The metabolic status and growth-rate patterns of these patients were within normal limits without alkaline therapy. All urine samples aspirated from the renal pelves were sterile. We conclude that a modified rectal bladder with a second ileal intussusception valve is the operation of choice whenever urinary diversion in children is indicated. PMID- 8731122 TI - Detubularized right colonic reservoir with intussuscepted ileal nipple valve or stapled ileal ("Lundiana") outlet. Clinical and urodynamic results in a prospective randomized study. AB - In a prospective randomized study, patients undergoing cystectomy and continent urinary diversion by means of a detubularized right colonic reservoir were randomized to one of two types of outlet: either an intussuscepted ileal nipple valve (n = 15) or a stapled ileal ("Lundiana") outlet (n = 15). There were no early complications from the reconstruction. Subsequently, one ileal nipple outlet required revision because of progressive catheterization difficulties. In the Lundiana group, perforation of the reservoir occurred in one case and a narrow stomal opening was revised in local anesthesia in two cases. Urodynamic assessment revealed similar pressures at rest for the two outlets. At stress (concomitant with reservoir contraction), there was a distinct increase in outlet pressure in the nipple valves, but this was rarely encountered in the Lundiana group. Questionnaires showed episodes of urinary leakage to be more common in the Lundiana group, whereas the reverse applied to catheterization difficulties. In both groups, however, the great majority of patients were satisfied or very satisfied with the function of the urinary tract. PMID- 8731123 TI - Continent diversion with the Mainz pouch. AB - From 1983 until July 1994, 561 patients in 2 urology departments (Mainz and Wuppertal) underwent a Mainz pouch 1 procedure. The Mainz pouch 1 was used for bladder augmentation in 60 patients, for orthotopic bladder substitution in 61 patients, and for continent cutaneous urinary diversion in 440 patients. In the group of continent cutaneous urinary diversion, the continence mechanism applied was an ileal intussusception nipple in 270 patients, an appendix stoma in 146 patients, a submucosal seromuscular bowel-flap tube in 14 patients, and a submucosal full-thickness bowel-flap tube in 10 patients. Indications for urinary diversion were bladder cancer in 339 patients, anatomical or functional loss of bladder capacity in 179 patients, and other primary or secondary malignancies of the bladder or true pelvis in 43 patients. After a mean follow-up period of 57 months (range, 3-127 months), early and late complications were encountered in 12% and 37% of the patients, respectively. In the bladder-augmentation group, 93% of the patients are completely continent day and night. All but three patients, who empty their reservoir by intermittent self-catheterization (CIC), void spontaneously by abdominal straining. In the orthotopic bladder-substitution group, 95% of the patients are continent during the daytime. To prevent urinary leakage, 13% have to empty their reservoirs regularly at 4-h intervals and 13% have to perform CIC to avoid residual urine. Among the patients treated with continent cutaneous urinary diversion, stoma failure occurred in 11%, stoma stenosis was encountered in 13% and required open revision in 2%, endoscopical incision in 10%, and conservative treatment (dilation) in 1% of cases. PMID- 8731124 TI - Present experience with the Indiana pouch. AB - The Indiana pouch was developed as a modification of the Glichrist procedure in 1984. The need for full detubularization of the reservoir was recognized early and was achieved by an ileal patch on the cecal reservoir or by an antimesenteric incision of the cecal segment with folding and transverse closure, the efferent limb and continence mechanism were modified to promote ease of catheterization and improved continence. These modifications gave markedly better results with lower reoperation rates. A group of 81 patients with a minimum of 2 years of follow-up are reported herein. The most recent modification uses absorbable GIA and TA55 staples to detubularize and close the reservoir. This stapling technique has been employed in 20 patients and has resulted in decreased operating times without any significant increase in complications. In its current form, the Indiana pouch meets all the criteria for a satisfactory continent cutaneous reservoir and has a favorable reoperation rate as compared with other pouches. PMID- 8731125 TI - The Mitrofanoff principle for continent urinary diversion. AB - The term Mitrofanoff describes a system for making a continent supra-public conduit into any reservoir for self-catheterisation. It requires a narrow tube, the commonest source of which is the appendix. Alternatively, the ureter, the Fallopian tube or a length of tailored intestine may be used. The tube is buried in the wall of the conduit in a tunnel about 5 cm long. There is no statistically significant difference in the results between any patient diagnosis, source of narrow tube or type of reservoir. About 90% of patients are continent. Up to 30% may have conduit complications, particularly stenosis at the skin level. Of the several systems of continent diversion available, the Mitrofanoff seems to be the easiest to learn and the most reliable. PMID- 8731126 TI - Appendiceal continence mechanisms in continent urinary diversion. AB - The creation of a safe, reliable, and easy-to-perform continence mechanism remains one of the most important problems during continent urinary diversion. The advent of the use of the appendix as an efferent segment brought through the umbilicus has greatly facilitated surgical procedures with very favorable results. Our experience with the insitu appendix as an efferent segment during continent cutaneous urinary diversion using the Mainz-pouch I technique over the past 6 years revealed a markedly decreased complication rate of 3.2% as compared with 7.2% in patients who received an ileocecal intussusception nipple. The routine use of the appendix as a continence mechanism during continent urinary diversion has proved to be a most valuable addition to our surgical armentarium. PMID- 8731127 TI - Evolution of outcomes with the ileal hydraulic valve continent diversion: reevaluation of the Benchekroun catheterizable stoma. AB - Initial reports with limited follow-up suggested that the ileal hydraulic valve was a satisfactory approach to continent urinary diversion that was associated with acceptable early complication rates and uniform efficacy in achieving continence. We sought to evaluate the later outcomes of patients with this form of continent urinary diversion. An average extended follow-up of 5 years was available on 11 patients who had undergone ileal hydraulic valve diversion. Although only 13% of patients had complications requiring surgical revision at 18 months of follow-up, at the 5-year follow-up, 91% had developed complications requiring surgical revision. Of these, 64% entailed major revision of the urinary diversion. The most common late complication was stomal stenosis (73%); calculus formation (36%) and devagination (36%) were also common. Although in some cases, stomal stenosis was successfully managed by superficial revision with skin flaps and grafts, most cases required complete replacement of the efferent limb. Use of the devaginated or stenosed ileal hydraulic valve to construct a flap-valve efferent limb based on the Mitrofanoff principal succeeded in salvaging the continent urinary diversions in these patients. Novel forms of urinary diversion require extended follow-up for accurate assessment of their feasibility and efficacy. Attentive surveillance with surgical revision as necessary in such cases is mandatory for the preservation of continence and renal function. PMID- 8731128 TI - Continent gastric pouch. AB - Between January 1985 and June 1995 a total of 12 patients (9 female, 3 male) underwent total reconstruction of the lower urinary tract using gastric tissue. Their mean age was 10 years (range, 5-25 years). Total gastric bladder substitution was performed in seven patients, whereas five other patients had composite continent reservoirs (stomach plus bowel) created. The diagnoses were cloacal exstrophy, classic bladder exstrophy, bladder rhabdomyosarcoma, bilateral ectopic ureters, and VACTERL association. The mean follow-up period was 4.5 years (range, 16 months to 9 years). The average bladder capacity was 309 ml, and the compliance averaged 12.9 ml/cmH2O. Continence was achieved in all patients, but the continence mechanism often required revision. Renal deterioration was noted only in one patient due to an obstruction at the site of a transureteroureterostomy. The complications included hyponatremic, hypochloremic metabolic alkalosis (two patients); prolapse of the neovagina (one patient); a ureterovesical junction obstruction (three patients); and revision of the neourethra or Mitrofanoff (six patients). Revision of the gastric tube used as the catheterizable channel was performed in three instances. Lengthening of the tube, including nippling in one instance, corrected persistent incontinence. Distal stenosis of a gastric tube required several plastic procedures for correction. A ureteral Mitrofanoff developed a distal stricture necessitating revision. Persistent leaking of an appendiceal Mitrofanoff was corrected with reimplantation. A tapered ileal neourethra required a tunneling procedure followed by Teflon injection to provide urinary continence. Complications seen most often involved the continence mechanism of these gastric neobladders or composite urinary reservoirs. Persistence, patience, and perseverance led to total urinary continence and adequate reservoirs for urine storage in all patients. PMID- 8731129 TI - Absorbable stapling techniques in continent urinary diversion. AB - The principle of bowel detubularization to decrease peristaltic pressure and increase reservoir capacity is applied in contemporary continent urinary diversions. The process of detubularization and refashioning of the spatulated bowel segment approximates 1 of operating time and is the most time-consuming aspect of pouch construction. The employment of devices applying absorbable staples ("absorbable staplers") has substantially reduced the time required to fashion bowel reservoirs. This article reviews the adaptation of the absorbable stapler to continent urinary diversion using small- and large-bowel segments. PMID- 8731130 TI - Dorsal laminectomy. PMID- 8731131 TI - Feline leprosy: spontaneous remission in a cat. AB - A two-year-old, male domestic shorthair with a solitary lesion of the right forelimb was presented for fine-needle aspiration biopsy of a suspected cutaneous, neoplastic process. Neutrophils, lymphocytes, and acid-fast bacilli packed in the cytoplasm of foamy macrophages and giant cells were seen on cytological examination. Bacteriological culture of the material from the skin lesion was negative for Mycobacterium spp. Intraperitoneal injection of homogenized material from the lesion resulted in generalized mycobacteriosis in one mouse after 10 months. Based on these results, a diagnosis of feline leprosy was made. No medical or surgical therapy was performed. Nonetheless, the lesion showed progressive and complete spontaneous remission within 3.5 months from the time of diagnosis; after 14 months, the cat still is free of disease. PMID- 8731132 TI - Autoantibodies to triiodothyronine and thyroxine in a golden retriever. AB - A golden retriever presented with signs of hypothyroidism occurring in conjunction with autoantibodies to both triiodothyronine (T3) and thyroxine (T4). The autoantibodies caused the apparent concentrations of total T3, total T4, and free T4 by analog assay to be high. However, free T4 concentration was nondetectable when measured using a dialysis assay. The dog's clinical condition markedly improved in response to L-thyroxine therapy, and the free T4 concentration by dialysis assay increased into the normal range. Thyroid hormone autoantibodies can confuse the diagnostic evaluation for suspected hypothyroidism. In dogs with autoantibodies to T4, measurement of free T4 by dialysis assay is useful for both diagnostic and therapeutic monitoring purposes. PMID- 8731133 TI - Hematological and biochemical changes and results of aerobic bacteriological culturing in dogs undergoing splenectomy. AB - Changes in complete blood count (CBC), bone marrow, reticulocyte count, coagulation profile, biochemical analysis, and serum iron, transferrin, and immunoglobulin M (IgM) concentrations were measured three and 10 days after splenectomy in 12 dogs. Spleens were cultured aerobically for bacteria and submitted for histopathological evaluation in 23 dogs undergoing splenectomy. There were no consistent changes in red blood cell (RBC), white blood cell (WBC), or platelet counts; bone-marrow samples; or biochemical profiles. Serum iron, transferrin, and IgM concentrations remained normal. Eight (35%) bacterial cultures yielded growth. Five of the 23 dogs had pyrexic episodes two-to-five days after surgery. In contrast to previous reports done on healthy dogs, this study shows that dogs with splenic disease have no characteristic changes in hematological or biochemical parameters after splenectomy. Rather, the changes tended to reflect the primary disease process. Splenic vascular compromise or a decrease in processing of bacteria may have resulted in the bacterial growth. There was no direct correlation to pyrexic postoperative episodes. PMID- 8731134 TI - Inadvertent prostatectomy as a complication of cryptorchidectomy in four dogs. AB - Inadvertent prostatectomy was identified as a complication of cryptorchidectomy in four dogs. Surgical correction of the resulting urethral trauma was attempted in each case. One case was euthanized due to a poor prognosis for recovery. Three cases survived without clinical evidence of urinary incontinence. Inadvertent prostatectomy during cryptorchidectomy may be avoided by adequate surgical exposure and proper identification of the abdominally retained testicle. PMID- 8731135 TI - Ophthalmic manifestations and complications of dental disease in dogs and cats. AB - Ophthalmic manifestations of dental disease may occur in dogs and cats because of the proximity between posterior maxillary teeth and the orbit. Ophthalmic disorders may be diagnosed promptly, but the initiating dental disease may be overlooked. Inappropriate diagnosis and treatment of dental disease may result in loss of teeth, irreversible vision-threatening ophthalmic disease, or, ultimately, loss of the globe. When ophthalmic examination results are suggestive of a primary dental disorder, thorough examination of the maxillary teeth is imperative to diagnose appropriately the underlying dental disease. Therapy should include treatment of the dental disease and secondary ophthalmic disorders. PMID- 8731136 TI - Treatment of canine acral lick dermatitis by behavior modification using electronic stimulation. AB - Canine acral lick dermatitis is characterized by excessive licking on areas of one or more limbs, usually near the carpus or tarsus. In this prospective study, five dogs with acral lick dermatitis were treated with remote punishment utilizing precisely controlled, momentary shock from an electronic training collar. The problem resolved in four dogs. Resolution was defined as one month in which no shocks (i.e., no electronic shock collar worn) or Elizabethan collars were utilized and no licking had occurred sufficiently to recreate a gross skin lesion. Relapse during the follow-up period of six-to-12 months occurred in two dogs, but licking stopped after brief retraining periods. PMID- 8731137 TI - Comparison of two antibarking collars for treatment of nuisance barking. AB - Two commercially available antibarking collars (i.e., a citronella spray collar and an electronic shock collar) were evaluated for efficacy and user satisfaction as reported by owners after a two-week trial period for each collar. While both collars were effective in decreasing barking (88.9% for the citronella spray collar and 44.4% for the electronic shock collar), most owners expressed a preference for the citronella spray collar. Owners perceived it as being more humane and indicated that they willingly would use it on a long-term basis to decrease their dogs' nuisance barking. While the citronella spray collar has some drawbacks, it is another tool for managing excessive barking. PMID- 8731138 TI - Ascites and hepatic cirrhosis in a cockatiel (Nymphicus hollandicus). AB - A mature, female cockatiel (Nymphicus hollandicus) was examined because of respiratory difficulties. Clinical and laboratory findings included ascites and evidence of hepatic disease (i.e., increased plasma bile acid concentrations, aspartate aminotransferase, and alkaline phosphatase activities). Plasma protein electrophoresis results were consistent with chronic-active inflammation. The albumin-to-globulin (A:G) ratio, calculated from plasma electrophoresis, was 0.3. Postmortem examination revealed severe hepatic fibrosis and a diffuse, interstitial, granulomatous lipid pneumonia. PMID- 8731139 TI - Longitudinal myotomy of the supraspinatus muscle: a new approach to the supraglenoid tubercle of the scapula. AB - A lateral approach to the shoulder joint via a longitudinal myotomy of the supraspinatus muscle was used in the surgical treatment of four cases involving avulsion fractures of the supraglenoid tubercle of the scapula. This approach was developed using cadaver specimens and was compared to previously reported approaches to the supraglenoid tubercle. This approach gave excellent, visualized exposure and permitted accurate reduction and fixation for this type of fracture. Excellent results were achieved in four clinical cases. PMID- 8731140 TI - Evaluation of fibular head transposition, lateral fabellar suture, and conservative treatment of cranial cruciate ligament rupture in large dogs: a retrospective study. AB - Sixty-one large dogs (weighing 22.7 kg or more) with cranial cruciate ligament ruptures (CCLRs) were treated with either fibular head transpositions (FHTs; n = 22 stifles), lateral fabellar sutures (LFSs; n = 39 stifles), or conservatively (CT; n = 11 stifles) with rest and aspirin. Based on owner evaluation, dogs treated with FHTs or CT did not perform as well as dogs treated with LFSs (p less than 0.05). There was no difference in owner evaluation scores for the dogs treated with FHTs or CT. Thirty dogs were reevaluated by investigators. No differences between treatment groups regarding age, sex, or time until diagnosis were noted. No differences in scores for lameness, stifle instability, or forceplate analysis among the treatment groups were observed. Degenerative joint disease progressed or remained severe regardless of treatment, based upon radiographic evidence. PMID- 8731141 TI - Canine mandibular osteosarcoma: 51 cases (1980-1992). AB - Fifty-one dogs treated for mandibular osteosarcomas (OSs) were studied retrospectively. Treatments were partial mandibulectomy (n = 32); partial mandibulectomy and chemotherapy (n = 10); partial mandibulectomy and radiation therapy (n = 3); partial mandibulectomy, radiation therapy, and chemotherapy (n = 4); and radiation therapy alone (n = 2). The overall one-year survival rate was 59.3%. Dogs treated with surgery alone had a one-year survival rate of 71%, which is higher than the one-year survival rate for dogs with appendicular OSs treated with surgery alone (p of 0.001 or less; hazard ratio of 0.29). There was no apparent effect of various treatment modalities, nor institution where treatment was given, nor histological type. Histological score and, to a lesser extent, histological grade were predictive of survival outcome. PMID- 8731142 TI - Chylothorax and chylous ascites in a dog with mediastinal lymphangiosarcoma. AB - A 10-year-old, male toy poodle presented for evaluation of a progressively worsening, harsh, nonproductive cough. Chylous pleural effusion, lymphopenia, thrombocytopenia, normoblastemia, and recurrent subcutaneous bruising were diagnosed. Surgical exploration of the cranial mediastinum revealed extensive, redundant connective tissue which was confirmed by histopathology to be mediastinal lymphangiosarcoma (LAS). During surgical placement of a fenestrated silastic mesh for passive pleuroperitoneal drainage, chylous ascites also was diagnosed. The patient was euthanized two days postoperatively due to persistent chylothorax. The etiopathogenesis of chylothorax and chylous ascites are discussed. Previous case reports of lymphatic endothelial neoplasia are reviewed. PMID- 8731143 TI - Similarity in behavior of polytetrafluoroethylene (ePTFE) prostheses implanted into different interfaces. AB - The biomaterial ePTFE is widely used in the clinical environment for vascular replacement or bypass, as well as in the repair of tissue defects, especially those involving the abdominal wall. The objective of this study was to evaluate the healing response to ePTFE prostheses implanted into a circulatory interface and a tissue interface, each in a different animal species. For vascular implants, the Sprague-Dawley rat (n = 60) was used, while the New Zealand white rabbit (n = 20) was used in the tissue replacement model. In the former, a vascular microprosthesis measuring 5 mm in length and 1 mm in internal diameter, having a porosity of 30 microns, was implanted into the common iliac artery. In the rabbit, a 7 x 5-cm fragment of ePTFE (Soft-Tissue Patch) was implanted into a defect in anterior abdominal wall that involved all the tissue layers. In this case, the prosthesis was left touching the intestinal loops. The implants were studied between 14 and 90 days of postimplantation by means of light microscopy, scanning electron microscopy, and immunohistochemistry. The latter involved the use of anti-rat (MAC-341) and anti-rabbit (RAM-11) macrophage-specific monoclonal antibodies. The behavior of the ePTFE in the different interfaces (vascular and abdominal wall) was similar with respect to the following aspects: the prosthesis presented a process of encapsulation which was more intense on the outer surface; colonization of the implant was limited to the outermost two thirds, with minimal invasion of the middle portion; colonization was absent on the edges of the prosthesis; collagenization of the interstice of the mesh occurred late; the foreign body reaction taking place on the outer surface was similar in both interfaces, with formation of a barrier consisting of macrophages and giant cells that did not penetrate the prosthesis; and, finally, in neither of the two models was vascular colonization of the PTFE prosthesis observed; rather, the angiogenic process was limited to the periprosthetic zones. The integration of the implant made of ePTFE is similar despite the differences in interfaces and the use of different animal species. The macrophage response does not determine the success or failure of the implant. PMID- 8731144 TI - Effects of indium and iron ions on in vitro calcium phosphate precipitation and crystallinity. AB - The elution characteristics of indium and iron ions from six kinds of dental casting alloys were studied in vitro. A certain amount of indium (0.01-1.70 mg/cm2) was solubilized from the surface of only one prosthetic semiprecious brand (Miro Bright) in either 1% lactic acid or 0.05% hydrochloric acid solutions. The elution of iron was below the detection limit or < 0.01 mg/cm2. We also studied the stimulatory effects of indium and ferric ions on the calcium phosphate precipitation in the absence and presence of an inhibitor (phosvitin, a phosphoprotein purified from egg yolks). Indium and ferric ions promoted the reaction, and their stimulatory effects were stronger than the inhibitory effects of phosvitin (250 micrograms/ml). These facts suggest that indium, but not iron, could be eluted into the mouth, and that the eluted indium may stimulate the formation of oral calcium phosphate precipitates, counteracting the inhibition by saliva-derived phosphoproteins. The effects of these metal ions on calcium phosphate precipitates were also studied using both X-ray diffractometry and infrared spectrophotometry. It was shown that both indium and ferric ions decreased the grade of crystallinity of the calcium phosphate precipitates (hydroxyapatite). PMID- 8731145 TI - Differential effects of different forms of hydroxyapatite and hydroxyapatite/tricalcium phosphate particulates on human monocyte/macrophages in vitro. AB - A possible complication associated with the use of hydroxyapatite (HA) or HA/tricalcium phosphate (HA/TCP) coating on the surfaces of prosthetic devices used for dental and orthopedic implants is their potential to fragment and thus exist as wear debris. In contrast to the so-called osteoconductive properties of HA or HA/TCP coatings, in particulate form these materials may lead to an adverse pattern of cellular and tissue responses at the bone-implant interface. We have established an in vitro cell culture system to characterize the biologic and biochemical effects of various particulate materials. The present study demonstrates that the HA/TCP particles derived from different sintering temperatures exhibit differential effects on cultured human monocyte/macrophages (M/M). The HA/TCP particles dried at 110 degrees C were the most biologically active, stimulating significant release of interleukin 1 beta (IL-1 beta), IL-6, tumor necrosis factor alpha, and prostaglandin E2 (PGE2), products implicated as important mediators of inflammation in diverse pathologic conditions. Other particles, sintered at either 900 or 1200 degrees C, did not stimulate production of cytokines or PGE2. HA/TCP particles from plasma-spray coatings also failed to release proinflammatory products. These results suggest that the biochemical and crystalline structural properties of particles markedly affects their capacity to modulate M/M function. This in vitro culture system should be useful in characterizing the specific physical and chemical properties of HA or HA/TCP particulates that are responsible for stimulating proinflammatory cell responses. PMID- 8731146 TI - Novel compliant and tissue-permeable microporous polyurethane vascular prosthesis fabricated using an excimer laser ablation technique. AB - A small-diameter vascular prosthesis with a multiply pored structure could have great potential to elevate the patency rate, for the following two reasons: 1) increased flexibility of the graft may increase compliance matching, consequently minimizing intimal hyperplasia; and 2) enhanced transmural tissue ingrowth may accelerate endothelialization. In this study, we fabricated a polyurethane (PU) based artificial graft with well-controlled micropores in terms of their diameter and distribution, which was achieved using a computer-aided excimer laser (KrF) ablation technique. Three types of microporous PU tubes (2 mm in internal diameter, 100 microns in wall thickness) were designed: pore size (100 microns) and longitudinal pore-to-pore distance (200 microns) were constant, and circumferential pore-to-pore intervals were 60 degrees (type 1), 30 degrees (type 2), and 15 degrees (type 3). The fabricated prostheses were coated with photoreactive gelatin, which was photogelled and chemically fixed on PU surfaces upon ultraviolet light irradiation. Scanning electron microscopy showed that pore size and arrangement were precisely controlled as designed, and that a gelatinous hydrogel layer was formed over the entire luminal surface. The stiffness parameter (beta), inversely related to compliance, was determined from the change in external diameter against intraluminal pressure. An increase in the number of pores around the circumference decreased the beta value. The type 3 graft, the stiffness parameter of which was very close to that of human coronary artery, was the most compliant among the three types. The combination of excimer laser directed microporing and photochemical surface processing techniques enabled the development of a novel compliant small-caliber vascular prosthesis, which is expected to show enhanced transmural tissue in growth in vivo. PMID- 8731147 TI - Canine bone response to tyrosine-derived polycarbonates and poly(L-lactic acid). AB - Tyrosine-derived polycarbonates are a new class of degradable polymers developed for orthopedic applications. In this study the long-term (48 week) in vivo degradation kinetics and host bone response to poly(DTE carbonate) and poly(DTH carbonate) were investigated using a canine bone chamber model. Poly(L-lactic acid) (PLA) served as a control material. Two chambers of each test material were retrieved at 6-, 12-, 24-, and 48-week time points. Tyrosine-derived polycarbonates were found to exhibit degradation kinetics comparable to PLA. Each test material lost approximately 50% of its initial molecular weight (Mw) over the 48-week test period. Poly(DTE carbonate) and poly(DTH carbonate) test chambers were characterized by sustained bone ingrowth throughout the 48 weeks. In contrast, bone ingrowth into the PLA chambers peaked at 24 weeks and dropped by half at the 48-week time point. A fibrous tissue layer was found surrounding the PLA implants at all time points. This fibrous tissue layer was notably absent at the interface between bone and the tyrosine-derived polycarbonates. Histologic sections revealed intimate contact between bone and tyrosine-derived polycarbonates. From a degradation-biocompatibility perspective, the tyrosine derived polycarbonates appear to be comparable, if not superior, to PLA in this canine bone chamber model. PMID- 8731149 TI - Preparation and characterization of high-surface-area polymer substrates for microcalorimetry. AB - The preparation and characterization of high-surface-area polymeric substrates suitable for the microcalorimetry of protein adsorption are described. High surface-area polystyrene, poly(styrene-co-butyl methacrylate) and poly(styrene-co allyl alcohol) were prepared by adsorbing polymer from solution onto fumed silica. Verification of adsorption of polystyrene by silica was determined by noting peak shifts of the surface silanol group in the infrared. The amount of polymer adsorbed was determined from adsorption isotherms. The minimum thickness of polystyrene required to mask silicon oxide properties was found to be that thickness at which contact angles became constant, about 35 A. Polymer densities were measured. Water contact angles on each polymer surface indicate that poly(styrene-co-allyl alcohol) has the surface most wettable by water. Polymer water interfacial energies were estimated from pendant drop results and a harmonic mean equation along with contact angles. Two methods were used to estimate the polar and dispersion components of the three polymers. Both methods predicted polystyrene to have the highest interfacial energy against water, and one method predicted poly(styrene-co-allyl alcohol) to have the lowest. A Wilhelmy plate study verified the change in interfacial properties as a function of contact time with water. A study of the heats of adsorption of lysozyme by each substrate using a modified Tien-Calvet microcalorimeter demonstrated the suitability of the substrates for microcalorimetry. PMID- 8731148 TI - Formation of apatite-collagen complexes. AB - An apatite-collagen complex was prepared in calcium beta-glycerophosphate solutions at pH 9.0 and 37 degrees C with the purpose of developing new bone substitutes that more closely resemble bone than currently available materials. Reconstituted type I collagen as well as sheet collagen were crosslinked in the presence of alkaline phosphatase and egg-yolk phosvitin. The crosslinked collagens were immersed in daily-renewed calcium beta-glycerophosphate solutions for 2 and 4 weeks to induce the deposition of apatite on the collagen fibers. After 2 weeks of reaction, for example, apatites deposited approximately two times the crosslinked collagen in weight. With reconstituted collagen, the complex showed some elasticity but no apatite was visually observed to detach under deformation with fingers and forceps. The complex, moreover, did not disintegrate when immersed in saline or animal blood. Nevertheless, the complex resorbed with no evidence of cytotoxicity when implanted in muscle tissues. These findings suggest that the apatite-collagen complex prepared would be useful as bone substitutes, especially for periodontal osseous lesion repair and alveolar ridge augmentation. PMID- 8731150 TI - Determination of contact phase activation by the measurement of the activity of supernatant and membrane surface-adsorbed factor XII (FXII): its relevance as a useful parameter for the in vitro assessment of haemodialysis membranes. AB - We investigated hemodialysis membrane biocompatibility with respect to contact phase activation by determination of FXII-like activity (FXIIA) on the membrane surface and in the supernatant phase, during plasma contact with various hemodialysis membranes using an in vitro incubation test cell. The results were compared to the influence of these membranes on the activation of purified FXII. A time course for the generation of activated FXII using purified FXII solution at physiologic concentrations on two similar negatively charged polymers was performed. The membranes assessed were regenerated cellulose (Cuprophan; Akzo Faser AG, Germany), modified cellulosic (Hemophan; Akzo Faser AG), acrylonitrile sodium methallyl copolymer-based membrane AN69S (Hospal, France), and SPAN, a new polyacrylonitrile-based copolymer (akzo Nobel AG). The plasma FXIIA at the membranes surface was significantly different between the membranes, while the supernatant phase FXIIA exhibited no significant differences. In contrast, activation of purified FXII in a plasma-free system with respect to supernatant activity indicated significant differences between the materials. A similar finding for the membrane-bound factor XIIA was also observed when purified factor XII was used. The membrane-bound FXIIA values observed in the plasma system containing heparin were significantly greater than in citrated plasma. This demonstrated the strong influence of heparin and the interaction of other plasma components to the membrane surface on the activation of contact phase of coagulation. PMID- 8731151 TI - Production of monoclonal antibodies to study corrosion products of CO-CR biomaterials. AB - Sensitivity to nickel, cobalt, and chromium is common among the general population. The identification of these sensitivities is generally by the detection of cell-mediated immunity. We have reported previously the use of an indirect enzyme-linked immunosorbent assay method to quantitate metal-specific antibodies in patients with total joint replacements. To study the haptenic potential of these metal ions, rabbit albumin-glutathione-metal complexes with chromium, cobalt, or nickel were injected into mice. The splenocytes from one mouse in each group which developed a strong antibody against GSH-metal complexes were isolated and fused with myeloma cells to produce monoclonal antibodies. Chromium, cobalt, and nickel antibodies had similar affinity and bound with the specific GSH-metal complex. There was very little cross-reactivity between these antibodies. An inhibition assay using these monoclonal antibodies was demonstrated to be a simple technique, suitable for quantitation of free metal in solution. PMID- 8731152 TI - Hydroxyapatite/metal composite coatings formed by electrocodeposition. AB - Early bone infusion by cementless fixation of composite orthopedic and dental implants consisting of metallic substrates and bioceramics is well documented. Calcium phosphate ceramics in general and hydroxyapatite (HA) in particular have been the most popular of the bioceramics used for coating metals. Here, a non line of sight coating procedure by electrocodeposition is reported for mechanically fixing HA particles in a metal matrix. Analyses of the coating showed excellent adhesion to the substrate and no structural transformation in either crystallinity or stoichiometry. Adhesion and surface coverage of HA depended upon the particle size. As a demonstration of the coating procedure's non-line of sight applicability, it was successfully used to coat titanium rods sintered with small titanium spheres. PMID- 8731154 TI - Characterization of a nitrogen-rich austenitic stainless steel used for osteosynthesis devices. AB - Two laboratory melts of the standardized nitrogen-rich austenitic stainless steel specified in ISO 5832-9:1992(E) have been characterized with respect to corrosion properties, mechanical properties, and microstructure. The two melts differ essentially in nitrogen concentration, namely, 0.42 and 0.46 wt %, respectively. Both melts were found to fulfill the requirements in the ISO standard for corrosion and mechanical properties. The resistance to pitting corrosion in a solution of 0.9% NaCl, intended to simulate the conditions in the human body, was demonstrated by a critical pitting temperature of about 70 degrees C for both alloys, which should be compared with 40 degrees C for the biocompatible reference steel AISI 316. While no difference in corrosion resistance was observed between the two alloys, a significantly higher mechanical strength and lower toughness were observed for the nitrogen-rich melt. Using electron diffraction Z-phase was identified in unaged material. These were present as primary precipitates, most likely precipitated in the liquid state owing to the high concentration of nitrogen in combination with the presence of the strong nitride former niobium. However, the influence of Z-phase on pitting corrosion is believed to be of minor importance. The ageing behavior was studied indirectly in terms of toughness as a function of ageing. Formation of the intermetallic phase- chi-phase--was observed, particularly during prolonged ageing at 800 degrees C. The total absence of chi-phase in forged bar condition shows that the cooling rates during production are sufficient to suppress the formation of chi-phase. PMID- 8731153 TI - Pharmacologic inhibition of particulate-induced bone resorption. AB - In this study, a rat calvaria/macrophage co-culture model was used to study the effects of various agents upon bone resorption induced by macrophage exposure to bone cement particles. The experimental group consisted of calvaria bone disks set in tissue culture medium on stainless-steel platforms into wells with macrophages adherent to the bottom which are exposed to the particles. Tumor necrosis factor alpha (TNF-alpha), prostaglandin E2 (PGE2), and calcium 45 (Ca45 were released in significant amounts in this system. Interleukin 1 alpha (IL-1 alpha) was not detected. Indomethacin inhibited the production of PGE2, but did not affect TNF release or inhibit the release of Ca45. Anti-TNF antibody neutralized the presence of TNF to undetectable levels, but did not affect PGE2 release or inhibit Ca45 release. The addition of calcitonin did not inhibit Ca45 release by calvaria. In contrast, the addition of disodium pamidronate, a member of the bisphosphonate family, was effective in inhibiting the release of Ca45 even after 96 h of incubation. In prior studies, incubation of calvaria in conditioned medium from macrophages exposed to cement particles led to resorption through a mechanism which is dependent upon TNF production by macrophages, and PGE2 production by cells in bone. In this two-way system, in which macrophages and cells in bone are allowed to interact, this dependency was no longer evident. Pamidronate was the only agent tested which suppressed the increase in bone resorption associated with macrophage exposure to bone cement particles to levels which were not significantly different from unexposed calvaria. By delaying or preventing bone resorption associated with macrophage exposure to bone cement particles, bisphosphonates may have a clinical role in cemented joint arthroplasty by decreasing the rate or incidence of aseptic loosening and prolonging implant longevity. PMID- 8731155 TI - Light-microscopic and electron-microscopic evaluation of short-term nerve regeneration using a biodegradable poly(DL-lactide-epsilon-caprolacton) nerve guide. AB - The aim of this study was to evaluate short-term peripheral nerve regeneration across a 10-mm. gap, using a biodegradable poly(DL-lactide-epsilon-caprolacton) nerve guide, with an internal diameter of 1.5 mm and a wall thickness of 0.30 mm. To do so, we evaluated regenerating nerves using light microscopy, transmission electron microscopy and morphometric analysis after implantation of 12-mm nerve guides in the sciatic nerve of the rat. Evaluation times ranged from 3-10 weeks. Three weeks after reconstruction, myelinated nerve fibers could be observed in the distal nerve stump. Ten weeks after reconstruction, the regenerating nerves already resembled normal nerves. In conclusion, we show that poly(DL-lactide epsilon-caprolacton) nerve guides can be successfully applied in the reconstruction of severed nerves in the rat model. Furthermore, we have observed the fastest nerve regeneration described thus far, after reconstruction using a biodegradable nerve guide. PMID- 8731156 TI - In vitro cell response to differences in poly-L-lactide crystallinity. AB - Many different processing techniques are currently being used to produce tissue regeneration devices from polyesters in the polylactide/polyglycolide family. While it is generally well recognized that processing techniques influence bulk mechanical and degradation properties of these materials, the effects on surface properties are relatively less well studied. We thus investigated the effects of processing conditions that are known to change bulk properties, but not composition, on the surface properties of poly-L-lactide (PLLA). Specifically, we investigated the role of bulk crystallinity of PLLA substrates on several physiochemical aspects of the surface and on the attachment, morphology, and differentiated function of cultured primary hepatocytes and growth of 3T3 fibroblasts. We fabricated smooth, clear PLLA films of 13-37% crystallinity. Glancing angle X-ray diffraction indicated that low crystallinity films lacked order in the first 50 A of the surface while relatively high crystallinity films had detectable order in this range. In other aspects, the surfaces of all PLLA substrates appeared identical with XPS, SEM, and advancing contact angle analysis, but contact angle hysteresis was slightly greater for more crystalline films. Although the physicochemical properties of the surfaces appeared almost identical, we observed differences in cell behavior on less crystalline versus more crystalline films. Hepatocytes formed spheroids on all PLLA substrates, but spheroid formation was faster (24-48 H) on crystalline substrates. quantitative image analysis was used to assess the average cell area as a function of time in culture, and our data confirm previous reports that retention of differentiated function is inversely related to cell spreading where function was assessed by P 450 enzyme activity. In addition, the growth rate of 3T3 fibroblasts was lower on crystalline substrates than on amorphous substrates. An important conclusion from this work is that processing techniques that lead to seemingly inconsequential changes in bulk and surface properties of these polymers may influence biological response. PMID- 8731157 TI - Effect of BaSO4 on the fatigue crack propagation rate of PMMA bone cement. AB - To determine the effect of BaSO4 on the fatigue crack growth rate, da/dN = C(delta K)n, of poly(methyl methacrylate) (PMMA) bone cement, radiopaque bone cement, radiolucent bone cement, and commercial PMMA (Plexiglas) were tested using a methodology based on ASTM E647. The crack growth rate of radiopaque bone cement was one order of magnitude less than that of radiolucent. Fractographic analysis showed that the regions of rapid catastrophic fracture were smooth for all materials tested. The radiopaque fatigue surface was rough and characterized by ragged-edged stacked plateaus, a morphology consistent with the model of crack propagation through the interbead matrix. Voids were visible in the interbead matrix on the order of the size of BaSO4 particles. The fatigue surface of radiolucent bone cement was relatively smooth, a morphology consistent with crack propagation through both the PMMA beads and interbead matrix. Fatigue striations were visible, and their spacing correlated well with crack propagation rates. The striations indicated an increased crack growth rate through the PMMA beads. PMID- 8731158 TI - Effect of biomaterial surface charge on the inflammatory response: evaluation of cellular infiltration and TNF alpha production. AB - A rat model was used to investigate the effect of net surface charge on polymer biocompatibility and its potential to modify and stimulate the inflammatory response. Poly(ether)urethane was taken as the base material and the net charge altered by introducing sulphonate ionic groups to the polymer backbone. Three differently charged poly(ether)urethanes were made with 10, 20, and 30% sulphonate substitution, giving a range of negative charge, with unmodified poly(ether)urethane used as a control. The polymers were implanted intramuscularly into rats for 2 days, and for 1, 2, and 12 weeks. After explantation, the cellular infiltration in the tissue surrounding the implants was evaluated using immunohistochemistry to stain for specific cell types: macrophages, neutrophils, lymphocytes, and the cytokine TNF alpha. In situ hybridization was used to detect expression of mRNA encoding TNF alpha. Stained sections were analyzed and the cellular response quantified using image analysis. Initially macrophages and neutrophils were observed around all the materials, but neutrophils were absent in all samples at 12 weeks. The 2-day time point had significantly more macrophages than the later time points. By 2 weeks the 20% charged polymer elicited significantly less neutrophil infiltration than the other three polymers. In all samples where macrophages were observed, cells staining positive fore TNF alpha protein and message also were observed. No T or B lymphocytes were observed in the infiltrates around the materials at any time point. The results indicate that surface charge can influence the early phase acute inflammatory response to an implanted material. PMID- 8731159 TI - Activation of the plasma coagulation system induced by some biomaterials. AB - The ability of some biomaterials to activate plasma coagulation system was examined in vitro. After contact of platelet-rich plasma with biomaterials, some markers of the thrombin formation, i.e., fragment 1 + 2 and fibrinopeptide A, and some inhibitors of the blood coagulation mechanism were tested. Fragment 1 + 2 and fibrinopeptide A were found to be increased by all of the materials, though to a different extent. In particular, fragment 1 + 2 and fibrinopeptide A were significantly increased upon contact with polybutylene terephthalate and with collagen coated polyethylene terephthalate, respectively. Also antithrombin III was shown to decrease following exposure to biomaterials, but statistical significance was found only for polyethylene terephthalate and polyvinylacetate. As a results of this wide range of variability in the parameters, it is advisable to explore the plasma coagulation system with a multiparametric approach in which thrombin formation and coagulation inhibitors are thoroughly investigated. PMID- 8731161 TI - Locomotor sensitization to [D-Trp(11)]neurotensin after repeated injections of the dopamine uptake inhibitor GBR12783 in rats. AB - Rats received one daily i.p. injection of the dopamine uptake inhibitor GBR12783 (1-[2-(diphenylmethoxy)ethyl]4-(3-phenyl-2-(propenyl)-piperazine) (10mg/kg) or vehicle for 9 days. Fourteen days after discontinuing treatment, their locomotor activity was assessed after injection of GBR12783 (5 mg/kg) or vehicle, then 6 days later, after i.c.v. injection of [D-Trp(11)]neurotensin (750 ng) or saline. A sensitization to the stimulant locomotor effects of both GBR12783 and [D Trp(11)]neurotensin occurred in rats exposed to the actimeter following the 1st, 5th and 9th injections of GBR12783. Rats without prior experience of the activity cages before the challenge tests showed no sensitization to either GBR12783 or [D Trp(11)] neurotensin. Our data suggest that a similar mechanism may underlie the locomotor sensitization to GBR12783 and the heterosensitization to [D Trp(11)]neurotensin. PMID- 8731160 TI - Nitric oxide induces cultured cortical neuron apoptosis. AB - A series of experiments were designed to examine the potential induction of apoptosis by nitric oxide (NO) donors on cortical neuronal cell culture. A 24 h exposure of three different NO donors, 3-morpholinosydnonimine, S-nitroso-N acetyl-penicillamine, and S-nitrosoglutathione, induced apoptosis as indicated by following histological (cellular and nuclear morphology) and biochemical markers (DNA oligonucleosomal fragmentation and protection by protein synthesis inhibitor). PMID- 8731162 TI - Effect of posture on the onset of fictive locomotion in the decerebrate rabbit preparation. AB - Spontaneous locomotor episodes were recorded from hindlimb muscle nerves of decerebrate curarized rabbit preparations. Changes in the static position of both hindfeet (from extended to flexed) or of the head (from horizontal to bent forward) were shown to elicit a shift of the first locomotor burst from flexion to extension. Interneurones whose activity was recorded in the lumbar spinal cord were active throughout the first locomotor burst only when the latter was an extensor burst. Such data show that proprioceptive inputs are able to determine the onset of central locomotor programmation. Neuronal interactions which, at the spinal level, could account for this effect, are discussed. PMID- 8731163 TI - Specific uptake of intracranial horseradish peroxidase (HRP) by microglial cells in the goldfish. AB - Intracranial injections of horseradish peroxidase (HRP) in the goldfish labeled a population of cells with many similarities to microglia. The fact that neither macroglia nor neurons appeared to be labeled by these injections supports this identification. The labeled cells responded to Wallerian degeneration of the optic paths by accumulating in the optic terminal zones, a cellular behavior which strongly supports their identification as microglia. PMID- 8731164 TI - Nitric oxide synthase activities are selectively decreased in vulnerable brain regions in thiamine deficiency. AB - Pyrithiamine-induced thiamine deficiency in the rat exhibits many neuropathological and biochemical similarities to Wernicke's Encephalopathy in human. Activities of constitutive nitric oxide synthase (NOS) were measured in vulnerable (thalamus and cerebellum) and non-vulnerable (hippocampus and striatum) brain regions of pyrithiamine-induced thiamine-deficient rats. NOS activities were significantly decreased in the thalamus (by 26%, P < 0.05) of presymptomatic thiamine-deficient rats compared to pair-fed controls. Following onset of symptoms, in addition to thalamus (-38%, P < 0.01), cerebellum (-50%, P < 0.01) also manifested significantly decreased activities of NOS. Hippocampal and striatal activities of NOS were unchanged at both presymptomatic and symptomatic stages of thiamine deficiency. Selectively decreased activities of neuronal NOS in the thalamus and the cerebellum extends the previous observations of region-selective metabolic changes and, ultimately, neuronal cell loss observed in thiamine deficiency. PMID- 8731165 TI - Protective effect of dichloroacetate in a rat model of forebrain ischemia. AB - Dichloroacetate (DCA) activates the pyruvate dehydrogenase complex (PDHC), and improves the recovery of cerebral pH, lactate, ATP, and PCr following reperfusion in animal models of forebrain ischemia. In order to determine whether this results in neuroprotection, rats were administered NaDCA (100 mg/kg or 10mg/kg i.v.) 10 min before 12 min of normothermic forebrain ischemia (bilateral carotid artery occlusion plus systemic hypotension, 45 mmHg). Neuronal injury assessed histopathologically 7 days post-ischemia was significantly reduced in the CA1 region of the hippocampus, the dorsal lateral striatum, and the neocortex, in rats treated with 100 mg/kg NaDCA, but not in rats treated with 10 mg/kg NaDCA. PMID- 8731166 TI - Vagus nerve mediates the increase in estrogen receptors in the hypothalamic paraventricular nucleus and nucleus of the solitary tract during fasting in ovariectomized rats. AB - The effect of total subdiaphragmatic vagotomy on estrogen-receptor immunoreactivity (ERIR) in the paraventricular nucleus (PVN) and nucleus of the solitary tract (NTS) was examined in fasted ovariectomized rats to clarify the peripheral inputs mediating fasting-induced increase in ERIR in these two nuclei. Vagotomy abolished the effect of 48-h fasting on the expression of ER in these two areas. The result indicates that the neural signal(s) that increase the expression of ER in the PVN and A2 region of the NTS following 48-h fasting is transmitted through the vagus. The involvement of the vagus in the fasting induced increase in ER in the PVN and A2 region may also be the same neural pathway involved in the suppression of pulsatile luteinizing hormone secretion in fasted female rats. PMID- 8731167 TI - 'Intact' dopaminergic midbrain neurons of the rat display unclamped dendritic Ca2+ currents. AB - Calcium channels play an important role in generating the complex electrophysiology of dopaminergic (DAergic) neurons in the substantia nigra pars compacta (SNc). To directly study these calcium currents in dendritically intact neurons, two issues needed to be addressed: the identity of the neuron (DAergic versus non-DAergic) and control of putative dendritic calcium conductances. A measure of the anomalous rectifier (Ih) was used to identify DAergic neurons. Both groups of neurons produced large (approximately 1 nA), sustained inward currents that persisted as 'plateau currents' for hundreds of milliseconds after the termination of the depolarizing voltage clamp step. We conclude that intact DAergic neurons can be identified under conditions optimal for recording calcium currents (i.e. Cs+ internal solution), but powerful dendritic currents limit rigorous biophysical analysis. PMID- 8731168 TI - Alteration of dynorphin and secretogranin II in the prolactin immunoreactive neurons of the rat lateral hypothalamus upon osmotic stimulation. AB - The prolactin immunoreactive neurons of the rat lateral hypothalamus were previously reported to express the dynorphin and secretogranin II genes. In the present study, the response of these neurons to osmotic challenge was immunocytologically investigated by using prolactin, dynorphin, secretogranin II and c-Fos antisera. In addition, the mRNA levels for secretogranin II and dynorphin were compared by in situ hybridization in controls and salt-loaded rats. For this model of chronic hyperosmolality, the prolactin and c-Fos immunoreactivities were not stimulated by salt drinking, but dynorphin and secretogranin II immunoreactivities as well as mRNA levels for dynorphin and secretogranin II significantly increased in the lateral hypothalamus. We suggest that the prolactin-immunoreactive neurons may be involved in the regulation of water homeostasis. PMID- 8731169 TI - In Syrian and European hamsters, the duration of sensitive phase to light of the suprachiasmatic nuclei depends on the photoperiod. AB - Light induction of the expression of Fos protein in the suprachiasmatic nuclei was used to investigate the photosensitive state of the clock in Syrian and European hamster kept under different photoperiods. We observed that the duration of the photosensitive phase is variable and tied to the length of the night. A maximal extension has been determined in both species studied. Finally, a 4 h lengthening of the phase of photosensitivity take approximately 3 weeks, while 3 days only are needed for its shortening. PMID- 8731170 TI - Memantine reduces functional and morphological consequences induced by global ischemia in rats. AB - In this study the effect of memantine, an antagonist at the N-methyl-D-aspartate receptor, on spatial learning deficit and on neuronal damage following transient cerebral ischemia was evaluated. Global ischemia was induced by four-vessel occlusion (4VO) for 20 min in rats. Memantine was administered 20 min before induction of ischemia at a dose of 10 or 20 mg/kg. One week after surgery spatial learning was tested in the Morris water maze. Treatment with the higher dose of memantine reduced the increase in escape latency and in swim distance induced by 4VO. Neuronal damage in the CA1 sector of the hippocampus and in the striatum produced by 4VO was significantly attenuated by 20 mg/kg memantine. Treatment with the lower dose of memantine had no influence on the deficit in spatial learning and the neuronal damage resulting from ischemia. The present data demonstrate that treatment with a neuroprotective agent like memantine can reduce functional as well as morphological sequelae induced by ischemia. PMID- 8731171 TI - Formalin induces biphasic activity in C-fibers in the rat. AB - While the formalin test is a widely used behavioral model of tonic chemogenic pain, little is known about the responses of primary afferent nociceptors to formalin. Formalin (2.5%, 50 microliters) was injected either directly in or adjacent to the mechanical receptive fields of single C-fibers isolated from the saphenous nerve of pentobarbital-anesthetized rats. The average formalin-evoked response in C-fibers (n = 29) over time was biphasic. This biphasic time course of the C-fiber response to formalin is similar to that of the behavioral response in the awake animal and is compatible with the hypothesis that increased C-fiber activity contributes to the behavioral response in phase 2, as well as in phase 1 of the formalin test. PMID- 8731172 TI - Capsaicin blocks tetrodotoxin-resistant sodium potentials and calcium potentials in unmyelinated C fibres of biopsied human sural nerve in vitro. AB - Topical application of capsaicin has been tested recently for treatment of painful peripheral neuropathy. In the present study, effects of capsaicin were explored on compound action potentials of isolated fascicles from human sural nerve biopsies. Capsaicin reduced the C fibre component by 30-60%; the remaining C fibres were not sensitive to the drug. A good correlation was found between the sensitivity of C fibres to capsaicin and their resistance to tetrodotoxin (TTX), i.e. C fibre action potentials recorded in the presence of TTX were completely blocked by capsaicin. Calcium action potentials seen after inhibition of axonal potassium conductances were also completely suppressed. The data indicate that application of capsaicin nearby human peripheral nerves might prevent action potential conduction in specific subtypes of C fibres. PMID- 8731173 TI - Accumulation of advanced glycation end products of the Maillard reaction with age in human hippocampal neurons. AB - The recent immunological demonstration of advanced glycation end products (AGE) of the Maillard reaction in several human tissues suggests a possible involvement of AGE in the aging process. We previously prepared a monoclonal anti-AGE antibody (6D12) which recognized N epsilon-(carboxymethyl)lysine. We examined, immunohistochemically, the effect of aging on AGE-immunoreactivity in hippocampal pyramidal neurons in ten brain tissue samples obtained at autopsy from subjects aged 20-85 years old. Using 6D12 antibody, our results demonstrated a positive correlation between AGE-immunoreactivity in hippocampal pyramidal neurons and age. A more intense immunoreaction was observed in the CA3-4 pyramidal neurons compared with that of the CA1 neurons, known to be vulnerable to the formation of neurofibrillary tangles. Our results suggest that AGE are probably involved in the aging process affecting the human central nervous system, and that AGE do not mainly contribute to the formation of neurofibrillary tangles, at least in the CA1 neurons. PMID- 8731174 TI - Dual termination modes of corticothalamic fibers originating from pyramids of layers 5 and 6 in cat visual cortical area 17. AB - Terminal morphology of corticothalamic fibers originating from cat area 17 was examined. Injections of an anterograde axonal tracer, phaseolus vulgaris leucoagglutinin (PHA-L), in area 17 resulted in labeling of small boutons in the dorsal lateral geniculate, perigeniculate, and thalamic reticular nuclei and in labeling of large boutons in the lateral nucleus of lateral posterior-pulvinar, ventral lateral geniculate, and pulvinar nuclei. Since it is well known that the dorsal geniculate nucleus is a major corticothalamic target for layer 6 pyramids and the lateral posterior-pulvinar complex is that for layer 5 pyramids, the findings indicate that layer 5 pyramids in cat area 17 project axons ending with large boutons, while layer 6 pyramids project those ending with small boutons. PMID- 8731175 TI - Enhanced dynamic complexity in the human EEG during creative thinking. AB - This study shows that divergent thinking, considered the general process underlying creative production, can be distinguished from convergent, analytical thought based on the dimensional complexity of ongoing electroencephalographic (EEG) activity. EEG complexity over the central and posterior cortex was higher while subjects solved tasks of divergent than convergent thinking, and also higher than during mental relaxation. Over the frontal cortex, EEG complexity was comparable during divergent thinking and mental relaxation, but reduced during convergent thinking. Results indicate that the basic process underlying the generation of novel ideas expresses itself in a strong increase in the EEG's complexity, reflecting higher degrees of freedom in the competitive interactions among cortical neuron assemblies. Frontocortical EEG complexity being comparable with that during mental relaxation, speaks for a loosened attentional control during creative thinking. PMID- 8731176 TI - Debrisoquine hydroxylase gene polymorphism frequencies in patients with amyotrophic lateral sclerosis. AB - Using PCR and restriction digest analysis, the frequencies of the variant cytochrome P450 debrisoquine hydroxylase CYP2D6 alleles CYP2D6(A) and CYP2D6(B) were investigated in 50 patients with amyotrophic lateral sclerosis (ALS) and 13 patients with ALS and frontotemporal dementia (FTD) and compared to those frequencies in patients with FTD alone and Alzheimer's disease (AD). The CYP2D6(T) allelic frequency was also assessed in ALS and ALS + FTD. Although the frequency of a poor metabolizer genotype was not increased in any disease group, there was a significant increase in the frequency of the CYP2D6(B) allele in the ALS patient group. This suggests that possession of a CYP2D6(B) allele may be a risk factor for the development of ALS, possibly conferring a 'gain of function' imposed by the mutation or reflecting linkage disequilibrium to a nearby susceptibility gene. PMID- 8731177 TI - Evidence that stress activates glial lactate formation in vivo assessed with rat hippocampus lactography. AB - Extracellular lactate of the rat hippocampus is inter alia increased by immobilization stress. The origin of lactate is, however, not well established, so it is not known whether it is mainly derived form neurons or glial cells. Dialysates were collected shortly (1 or 2 days) or with a delay (14 or 15 days) after implantation of the probe. In the short-term experiment lactate increased after stress, both with or without glucose added to the perfusate. In the long term experiment there was marked gliosis around the dialysis probe and the stress effects were seen only in the presence of 5 mM glucose. The results are consistent with the idea that stress induces glycogenolysis and lactate export from astroglial cells via neurotransmitter or hormonal related processes. PMID- 8731178 TI - Polycystic ovary syndrome. PMID- 8731179 TI - Controlled ovarian stimulation using highly purified FSH results in a lower serum oestradiol profile in the follicular phase as compared with HMG. AB - We have examined the efficacy of highly purified follicle stimulating hormone (FSH-HP) for controlled ovarian stimulation in our in-vitro fertilization (IVF) programme, and compared the results obtained with this preparation with those using human menopausal gonadotrophin (HMG) in 15 patients who had received treatment with both FSH-HP and HMG in consecutive cycles (n = 39). No differences were found in the duration of stimulation, which was 13.9 days (HMG) as compared with 14.3 days (FSH-HP). However, in the FSH-HP-treated cycles we found a striking difference in the rise of serum oestradiol, which was significantly lower than in HMG-treated cycles (2953 +/- 938 pmol/l as compared with 6349 +/- 3683 pmol/l on the day before ovum retrieval). Number and size of follicles were similar in the two groups, as were oocyte characteristics. Increase in endometrial thickness at two days prior to ovum retrieval was slightly higher after HMG. The results indicate that in combination with a long gonadotrophin releasing hormone agonist (GnRHa) protocol, pure FSH is sufficient for adequate follicle recruitment and growth. However, since FSH-HP resulted in markedly reduced concentrations of serum oestradiol as compared to HMG cycles, IVF programmes using repeated oestradiol measurements to decide the day of ovum retrieval must take this into consideration in order not to prolong the stimulation unnecessarily. PMID- 8731180 TI - Weekly duration of dialysis treatment--does it matter for survival? PMID- 8731181 TI - Identifying the genes for human hypertension. PMID- 8731182 TI - Atypical presentation of systematic nocardiosis and successful treatment with meropenem. PMID- 8731183 TI - Third International Conference on Haemorrhagic Fever with Renal Syndrome (HFRS) and Hantaviruses. PMID- 8731184 TI - What is new in CAPD? (highlights of the 7th Congress of the International Society of Peritoneal Dialysis, June 1995). PMID- 8731185 TI - Pathogenesis of chronic renal failure in the primary glomerulopathies, renal vasculopathies, and chronic interstitial nephritides. PMID- 8731186 TI - Significance of tubulointerstitial changes in diabetic renal disease. PMID- 8731187 TI - Apoptosis and renal scarring. PMID- 8731188 TI - Modulation of glomerular sclerosis and interstitial fibrosis by native and modified lipoproteins. PMID- 8731189 TI - Phenotypic modulation of renal cells during experimental and clinical renal scarring. PMID- 8731190 TI - Phenotypic features of cortical interstitial cells potentially important in fibrosis. PMID- 8731191 TI - The underlying cellular mechanism of fibrosis. AB - Fibrosis is a common sequela of various exogenous insults to a variety of parenchymal tissues. The underlying mechanisms of the induction and progression of fibrosis both at the molecular and cellular level have not been clarified so far. In the present study the cellular processes that ultimately may lead to interstitial fibrosis are described using the model of radiation-induced terminal differentiation in the fibroblast/fibrocyte cell system. The data reported herein will provide evidence that exogenously induced changes in the proliferation and differentiation pattern of the fibroblast/fibrocyte cell system based on either autocrine and/or paracrine mediators represent the underlying cellular mechanism of fibrosis. Using co-culture systems of parenchymal cells (fibroblasts and type II pneumocytes), the intercellular communication via cytokines, which may lead to fibrosis have been studied. TGF beta 1 could be described as one key modulator of these cellular processes. PMID- 8731192 TI - Possible pathogenesis of renal fibrosis. PMID- 8731193 TI - Transdifferentiation of hepatic stellate cells (Ito cells) to myofibroblasts: a key event in hepatic fibrogenesis. AB - Hepatic stellate cells (HSC) are the major source of extracellular matrix components (ECM), if these precursor cells are activated in areas of necroinflammation to proliferate and to transdifferentiate from the retinoid storing to the ECM-producing phenotype termed myofibroblast (MFB). The mechanisms of HSC activation are crucial for the understanding of liver fibrogenesis. Activation is promoted by cytokines from Kupffer cells (KC) and platelets, of which TGF-beta is of great importance. It stimulates in HSC gene expression of ECM molecules but inhibits proliferation and induces phenotypic transdifferentiation. Latent TGF-beta is activated in cooperation with endothelial cells. TGF-alpha, also secreted by KC, stimulates HSC proliferation. In addition, hepatocytes (PC) enhance proliferation of HSC but ECM production is not affected. The mitogenic effect is mediated by TGF-alpha, IGF-1 and other cytokines. The paracrine signals of PC are increased by PC damage. During transdifferentiation MFB increasingly express TGF-beta and TGF-alpha, which are suspected to be involved in autocrine stimulation of MFB and paracrine activation of still untransformed HSC. A three-step cascade model of HSC activation is suggested. A preinflammatory phase initiates HSC activation by discharge of mitogenic cytokines (TGF-alpha, IGF-1) from damaged PC followed by the inflammatory phase based on cytokines (TGF-beta, etc.) from activated KC. In the postinflammatory phase MFB are stimulated by autocrine mechanisms contributing potentially to a perpetuation of the fibrogenic process even after cessation of the primary event (PC damage). PMID- 8731194 TI - Role of the myofibroblast in pulmonary fibrosis. PMID- 8731195 TI - Expression of genes that promote renal interstitial fibrosis in rats with proteinuria. AB - The development of progressive tubular atrophy and interstitial fibrosis represents a final common pathway leading to renal insufficiency. My laboratory has been investigating several rat models of primary glomerular disease in an effort to determine cellular and molecular mechanisms of renal interstitial fibrosis. These models include puromycin aminonucleoside nephrosis (PAN), protein overload proteinuria, passive Heymann nephritis, and diet-induced hypercholesterolemia. From a functional perspective, it is likely that the associated loss of tubules accounts for the decline in renal function. Four recurrent themes are emerging from our studies. PMID- 8731196 TI - Proteoglycans: their possible role in renal fibrosis. PMID- 8731197 TI - Matrix metalloproteinases (MMPs) in renal fibrosis. PMID- 8731198 TI - Signaling from the matrix to the cytoskeleton: role of cell surface proteoglycans in matrix assembly. PMID- 8731199 TI - Extracellular matrix remodeling and the regulation of epithelial-stromal interactions during differentiation and involution. AB - An intact basement membrane is essential for the proper function, differentiation and morphology of many epithelial cells. The disruption or remodeling of the basement membrane occurs during normal development as well as in the disease state. To examine the importance of basement membrane during development in vivo, we altered the matrix metalloproteinase and tissue inhibitor of metalloproteinases balance in mammary gland. Inhibition of matrix metalloproteinase synthesis by glucocorticoids or implants or transgenic overexpression of tissue inhibitor of metalloproteinases -1 delays matrix degradation and the involution process after weaning. The mammary glands from transgenic mice that inappropriately express auto-activating isoforms of stromelysin-1 are both functionally and morphologically altered throughout development. Transgenic mammary glands have supernumerary branches, and show precocious development of alveoli that express beta-casein expression and undergo unscheduled apoptosis during pregnancy. This is accompanied by progressive development of an altered stroma, which becomes fibrotic after postweaning involution, and by development of neoplasias. These data suggest that metalloproteinases and disruption of the basement membrane may play key roles in branching morphogenesis of mammary gland, cell cycle, apoptosis, and stromal fibrosis as well as in induction and progression of breast cancer. PMID- 8731200 TI - Prevention of progression of renal fibrosis: how far are we? PMID- 8731201 TI - The effect of ACE inhibitors on the expression of matrix genes and the role of p53 and p21 (WAF1) in experimental renal fibrosis. AB - Tubulointerstitial fibrosis in unilateral ureteral obstruction (UUO) is driven by increased levels of angiotensin II (AII). In this study we administered the angiotensin converting enzyme (ACE) inhibitor enalapril to rats with UUO after tubulointerstitial fibrosis was established. Treatment with the ACE inhibitor halted the progression of fibrosis and to a significant extent reversed some aspects of tubulointerstitial fibrosis. It is suggested that enalapril administration may be an effective means of preventing the progression of tubulointerstitial fibrosis. PMID- 8731202 TI - Interleukin-1 in renal fibrosis. PMID- 8731203 TI - Staphylococcus epidermidis adhesion to films deposited from hydroxyethylmethacrylate plasma. AB - The adhesion of S. epidermidis ATCC 35984 strain on polystyrene (PS) disks coated by films deposited from hydroxyethylmethacrylate (HEMA) plasma was evaluated and compared to adhesion on untreated PS and oxygen-plasma-treated PS. Films were deposited keeping constant the monomer flow rate while the discharge power ranged from 40-100 W in order to obtain coating with different surface properties. Surface chemistry, energetics, and morphology were evaluated by Electron Spectroscopy for Chemical Analysis (ESCA), contact angle measurement, and Atomic Force Microscopy (AFM), respectively. Bacteria adhered more to the plasma deposited or plasma-treated surfaces than to untreated PS, but no significant difference was recorded among the samples obtained using different deposition conditions. According to the surface energetic analysis, plasma-deposited and plasma-treated surfaces bear a strong Lewis-base character, so it is possible to hypothesize a marked contribution of electron donor-electron acceptor interactions to the mechanism(s) controlling adhesion between synthetic and bacterial surfaces. PMID- 8731204 TI - A new biological glue from gelatin and poly (L-glutamic acid). AB - This study describes the potentiality of hydrogels composed of gelatin and poly(L glutamic acid) (PLGA) as a biological glue for soft tissues and compares its effectiveness with that of a conventional fibrin glue. Water-soluble carbodiimides (WSC) were used to crosslink the aqueous mixture of gelatin and PLGA. The mixed aqueous solution of gelatin and PLGA set to a hydrogel by use of WSC as rapidly as BOLHEAL fibrin glue. An addition of PLGA to gelatin aqueous solution reduced not only its gelation time but also the WSC concentration necessary for hydrogel formation. The cured hydrogel exhibited firm adhesion to the mouse skin and other soft tissues with a higher bonding strength than BOLHEAL fibrin glue. Cohesive failure in the hydrogel was observed when the gel-tissue bond was broken, in contrast to BOLHEAL fibrin glue. The bonding strength of the gelatin-PLGA hydrogel became higher with the increasing PLGA concentration. The inflammatory reaction around the gelatin-PLGA hydrogel subcutaneously implanted in mice was mild, and the hydrogel was gradually absorbed with time in vivo. A toxicity test demonstrated that the concentration of WSC necessary as a biological glue was low enough not to induce its toxicity. PMID- 8731205 TI - Numerical and experimental stress analysis of a polymeric composite hip joint prosthesis. AB - A comparative stress analysis of a polymeric composite hip joint replacement was performed. A prototype short carbon-fiber reinforced PEEK (CF/PEEK) prosthesis was manufactured by injection molding. Finite element (FE) analysis was conducted on intact femurs and femurs fitted with the CF/ PEEK and the titanium prostheses under various loading conditions. FE models were validated by experimental strain gauge measurements by using synthetic femurs. There was a good agreement between the two methods except in the hoop strain of the femur in the calcar region because of the assumption of the isotropic material properties. The stem stresses were lower for the CF/PEEK prosthesis than for the titanium prosthesis. The maximum stress was in the spigot of the CF/PEEK prosthesis, but in the middle third of the stem of the titanium prosthesis. Stress generated in the cement was almost equal for both prostheses although more load was transferred, via cement, to the femur with the CF/ PEEK prosthesis because the load transfer took place over a larger area. An out-of-plane component of the joint load causes higher prosthesis and cement stresses. PMID- 8731206 TI - Assessment of metal extract toxicity on human lymphocytes cultured in vitro. AB - In this study the toxic effects of chromium, nickel, and cobalt extracts on in vitro cultured lymphocytes were evaluated. Graphite furnace atomic absorption spectrometry was used to measure the ion concentration. After serial dilution of the extracts, the viability of lymphocytes at 24, 48, and 72 h was estimated by flow cytometry, including propidium iodide staining and light scatter property assessment, and by MTT reduction test. The results of the investigation allowed us to conclude that 1) standardization of the procedure for preparing extracts is fundamental to obtaining repeatability of results; 2) the toxicity of an extract cannot be evaluated with a single viability assay; a combination of functional and structural tests is required; 3) when methods based on enzymatic reactions are performed, e.g. MTT test, it is advisable to replace the extract containing metal ions with fresh medium in order to avoid any interference with viability testing; 4) the amount of Co and Ni in the extract is similar, but the Cr release is very poor; 5) the lower toxicity of Cr extract probably is due to the lower ion concentration; 6) the assessment of 50% cytotoxic concentration (TC50) allows quantification of materials toxicity and comparison of various metals; and 7) the determination of a noncytotoxic concentration, i.e., a concentration lower than TC10, is required for subsequent investigation of cell functions because such studies can be carried out only on viable cell population. PMID- 8731207 TI - Incorporation of a fluorocarbon polymer implanted at the posterior surface of the rabbit cornea. AB - An implant of porous expanded polytetrafluoroethylene (PTFE) in the corneal stroma allows fast cell colonization and can become translucent. We studied the behavior of the same polymer implanted in the anterior chamber of the rabbit eye and sutured to the posterior surface of the cornea. The expanded tetrafluoroethylene was provided as 200-micron thick sheets in 3 pore diameters (20, 50 and 80 microns). Disks (5 mm) were implanted in the anterior chambers of 20 rabbits and sutured to the posterior surface of the cornea. Histological and ultrastructural studies were performed after 3 and 4 months. Quantimetry was done on TEM images to analyze the fibrillar structure of the intercellular matrix inside and around the implanted polymer. The material was well tolerated. In all cases there was mild central corneal edema, which disappeared after 1 month. Mild neovascularization occurred in five cases, decreasing after 1 month. The polymer became translucent after 8 days. Keratocytes from the corneal stroma colonized the implant via breaks in the Descemet's membrane along the sutures. Quantimetry showed three types of fibrils inside and beside the polymer. Corneal endothelial cells regenerated over the fibroblasts and the polymer. This fluorocarbon polymer implanted in the anterior chamber and sutured to the posterior surface of the cornea was well tolerated, and there was real incorporation, with keratocytes producing collagen fibrils inside the polymer and endothelial cells forming a posterior cell monolayer. This is promising for the development of a keratoprothesis with posterior fixation. PMID- 8731208 TI - Calcification of polyurethanes implanted subdermally in rats is enhanced by calciphylaxis. AB - Calcification complicates the use of the polymer polyurethane in cardiovascular implants. To date only costly experimental circulatory animal models have been useful for investigating this disease process. In this paper we report that polyurethane calcification in rat subdermal implants is enhanced by overdosing with a vitamin-D analog. The calcification-prone state, known as calciphylaxis, was induced in 4-week old rats by oral administration of a vitamin-D analog, dihydrotachysterol. We studied two commercially available polyurethanes (Biomer and Mitrathane) and two proprietary polyurethanes (PEU-2000 and PEU-100). PEU-100 is unique because it is derivatized with ethanehydroxy-bisphosphonate (EHBP) for calcification resistance. Polyurethane calcium and phosphate levels and morphological changes due to calciphylaxis were compared with those of control rat subdermal explants in 60-day studies. Increased polyurethane mineralization was observed due to calciphylaxis with 60-day rat subdermal explants of Biomer, Mitrathane, and PEU-2000 (calcium levels, respectively, 4.13 +/- 0.56, 18.61 +/- 2.73, and 3.37 +/- 0.22 microgram/mg, mean +/- standard error) as compared to control explants (calcium levels, respectively, 1.22 +/- 0.1, 12.57 +/- 0.86, and 0.20 +/- 0.86 microgram/mg). The study also demonstrated that with 60-day implants calciphylaxis had no side effects on somatic growth and serum calcium levels. Explant surface morphology of these polyurethane explants examined by scanning electron microscopy, back scattering electron imaging coupled with energy dispersive X-ray spectroscopy, and light microscopy demonstrated the presence of predominantly surface-oriented calcification. PEU-100, derivatized with 100 n.moles/ mg of EHBP, resisted calcification with explant calcium levels 0.51 +/- 0.01 (calciphylaxis) and 0.38 +/- 0.01 (control) microgram/mg. It is concluded that calciphylaxis enhances superficial polyurethane calcification in rat subdermal implants and that an EHBP-modified polyurethane resists calcification despite calciphylaxis. Rat subdermal implants using calciphylaxis may be generally useful for evaluating the calcification potential of various biomedical polymers. PMID- 8731209 TI - Effects of titanium on transcriptional and post-transcriptional regulation of fibronectin in human fibroblasts. AB - The effects of commercially pure titanium (Ti) on the regulation of fibronectin gene expression and synthesis were investigated in early-passage human gingival fibroblasts. The fibroblasts were cultured on 50 nm Ti-coated silicon wafers treated with radio-frequency glow discharge prior to use and on Falcon tissue culture plastic (TCP) dishes as a control. Northern hybridization analysis revealed that fibroblasts cultured on Ti reduced the fibronectin mRNA level by 58% at 16 h, but increased it by 2.6-fold at 90 h, although the cell numbers and house-keeping gene GAPD mRNA levels on these two surfaces were essentially the same. The amount of total RNA was slightly less on the Ti surface. While the total [35S]methionine incorporation was essentially unaltered, the amount of [35S]methionine-labeled fibronectin was significantly increased in cells cultured on a Ti surface in early cultures but decreased in the late cultures. The apparent discrepancy between the increased fibronectin mRNA levels and decreased translation could be explained by a 30% reduction in fibronectin mRNA half life in cells cultured on Ti. The distribution of fibronectin between the medium and the cell layer also was altered on Ti surfaces, with a approximately 100-fold increase of fibronectin assembled in extracellular matrix at 16 h, but a 36% reduction at 90 h. In contrast, the amount of fibronectin recovered in the medium was essentially unchanged. The total amount of protein assembled into the extracellular matrix by cells on Ti increased 2.1-fold at 16 h but decreased by 19% in 90-h cultures. These significant changes in fibronectin gene activity and gene product distribution by cells cultured on Ti surfaces demonstrate that the surface chemistry of biomaterials can selectively regulate the cellular behavior at the molecular level and, conversely, that molecular biological techniques provide sensitive indicators of the molecular biocompatibility of implant materials. PMID- 8731210 TI - A study of the influence of biphase calcium phosphate ceramics on bacterial strains: in vitro approach. AB - An in vitro study of the influence of Biphase Calcium Phosphate Ceramics (BCPCs) on bacterial strains is presented. The test procedures were carried out in saline test solution to which different quantities (2,4,50,100, and 200 mg) of BCPCs(samples A and B) were added. The influence of BCPCs on standard bacterial strains has been determined by measuring bacterial contamination using the plate count method. The testing method was validated with membrane filtration and direct inoculation in culture media of sediment and supernatant. The test demonstrated reduction of the bacterial cell population in from 1 to 24 h in all experiments. The antibacterial effect revealed the specific inherent properties of the BCPCs under investigation. PMID- 8731211 TI - Local accumulation of titanium released from a titanium implant in the absence of wear. AB - Titanium and its alloys, like the majority of metallic implant materials, release passive metal dissolution products. This raises the issues of amount and fate, i.e. transport, storage, and/or excretion of these metal dissolution products. In this paper we document titanium levels in tissues local to a commercially pure titanium implant in the absence of wear; compare these values to control tissues; and determine the relative contribution of the local accumulation to total release. Titanium fiber felts were implanted into the tibia of rabbits for periods up to 1 year. Bone and muscle tissue samples near the implant were collected. Using electrothermal atomic absorption spectrophotometry the samples were analyzed for titanium content. Compared to controls, titanium levels in the bone near the implant were elevated at 1-, 4-, and 12-month postoperative time points. The 12-month time point had higher periprosthetic bone titanium levels than both the 1- and the 4-month implant groups. Titanium levels in muscle tissue near the felt also indicated release was occurring. The data support the hypothesis that metal species released from titanium implants in the absence of wear have a limited solubility. As a result, they tend to remain in an area local to the implant. PMID- 8731212 TI - Surface microarchitectural design in biomedical applications: in vitro transmural endothelialization on microporous segmented polyurethane films fabricated using an excimer laser. AB - We describe the preparation of segmented polyurethane (SPU) films with round micropores and present a quantitative assay method of endothelial cell (EC) migration through micropores of and growth on microprocessed SPU films as an in vitro model of transmural endothelialization in open-cell-structured small diameter vascular grafts. The micropored films, pores of which ranged from 9 to 100 microns in diameter, were microfabricated using an excimer laser. Time dependent processes of EC ingrowth through micropores of SPU films with different pore sizes, which have a confluent monolayer sheet on one face and are cell free on the other, and subsequent endothelialization were quantitatively studied. The circular cellular sheet centered at the micropores expanded as incubation proceeded. Markedly retarded migration was found for the smallest pore size (9 microns in diameter). The larger the pore, the higher was the endothelialization rate. The endothelialization characteristics were studied on multiply microspored films of different pore sizes and densities, each of which was prepared so as to provide a fixed total pore area per unit area (0.01 mm2 per mm2). The highest endothelialization rates in an early incubation period were found on films with microspores between 18 and 50 microns in diameter. PMID- 8731213 TI - Quantitative assessment of apatite formation via a biomimetic method using quartz crystal microbalance. AB - Quantitative assessment of hydroxyapatite formation on a gold surface via the biomimetic method, composed of a nucleation step in a simulated body fluid (SBF) containing glass powders and a subsequent apatite growth step in glass powder free SBF, was made using a quartz crystal microbalance (QCM) technique. The frequency change of the QCM linearly increased with increasing soaking time, and largely depended on the nucleation period. The growth rates, defined as daily increase in thickness, increased monotonicly with an increasing nucleation period of up to 96 h, thereafter being constant at 2.0 microns/day. The growth rate of the apatite layer increased with increasing temperature of the SBF: 0.9, 2.0, and 3.8 microns/day at 25, 37, and 50 degrees C, respectively. The Arrhenius-type activation energy for the growth of apatite was 47.3 kJ/mol. The QCM method was found to be a very powerful tool for quantitative, in situ measurement of precipitation and growth of apatite in real time. PMID- 8731214 TI - Synergistic induction of tumor necrosis factor alpha by bacterial lipopolysaccharide and lipoteichoic acid in combination with polytetrafluoroethylene particles in a murine macrophage cell line RAW 264.7. AB - The induction of tumor necrosis factor alpha (TNF-alpha) by polytetrafluoroethylene (PTFE) particles (5-50 microns) and by bacterial lipopolysaccharide (LPS) and lipoteichoic acid (LTA) was examined in RAW cell cultures. Twenty-four-hour culture supernatants from the treated and control cells were assayed for TNF-alpha using a mouse L929 cell cytotoxicity assay. Untreated RAW cells produced low levels of endogenous TNF-alpha in the culture supernatants. Addition of 0.5 ng to 1 microgram/ mL LPS or 1 ng to 1 microgram/ml LTA increased the TNF-alpha production by 7-3570-fold and 2-815-fold, respectively. Addition of 1-5 mg PTFE increased the TNF-alpha production by 6-17 fold over the untreated control cell levels. The cells exposed to PTFE and 0.5 ng/mL LPS or 5 ng/mL LTA produced TNF-alpha levels that were significantly higher than those produced by any inducer alone. Thus, both LTA, a Gram-positive bacterial cell wall component and LPS, a Gram-negative bacterial cell wall component, can induce TNF-alpha production, which is further enhanced by PTFE particles in RAW cells. PMID- 8731215 TI - Comparison and quantitation of wear debris of failed total hip and total knee arthroplasty. AB - To compare the physical properties of debris particles associated with failed total hip and total knee arthroplasty, we applied a recently developed assay to electronically characterize the size, number, and composition of debris particles isolated from tissues adjacent to failed implants. We identified 21 samples (from 20 patients) of hip synovia and 35 samples (from 32 patients) of knee tissues that had been obtained at the time of revision arthroplasty. There were 12 females and 9 males in the hip group, and 16 females and 19 males in the knee group. Primary arthroplasty was performed for osteoarthritis (OA, 15 cases) or rheumatoid arthritis (RA, 6 cases) in the hip, and for OA (23) or RA (12) in the knee. Patients ranged in age from 23 to 85 (mean 59 years) for total hip, and from 27 to 84 (mean 61 years) for total knee arthroplasty. Implantation duration was from 5 to 123 months (mean 37.8) for total hip, and from 11 to 123 months (mean 63.1) for total knee arthroplasty. All of the implants were composed of cobalt-chromium alloy articulating with ultrahigh-molecular-weight polyethylene. The number of particles smaller than 10 microns ranged from 1.04 x 10(8)/g to 1.91 x 10(10)/g in the hip, and from 6.69 x 10(8)/g to 2.13 x 10(10)/g in the knee. Energy-dispersive X-ray spectroscopy and polarized light analysis showed both polyethylene and metal particles in most cases. The mean diameter of particles smaller than 10 microns was 0.72 +/- 0.2 microns in the hip, and 0.74 +/- 0.1 microns in the knee. Evaluation of particles larger than 10 microns showed a larger range of particle size in knee tissues (maximum 6.1 mm, mean 283 microns), than in the hip tissues (maximum 826 microns, mean 81 microns) (p < 0.001). Very small particles are common in both groups, but it appears that a larger range of particle sizes is present adjacent to failed knee than to failed hip prostheses. The higher frequency of large particles in failed knee prostheses probably reflects the perceived higher rate of delamination and fragmentation of tibial and patellar compared to that of acetabular polyethylene. PMID- 8731216 TI - Conjugates of insulin with copolymers of N-(2-hydroxypropyl) methacrylamide: effects on smooth muscle cell proliferation. AB - The hypothesis that an elevated plasma insulin level contributes to an increase in coronary heart disease has led to studies of the mitogenic effect of native insulin and its conjugates on smooth muscle cells (SMC). In this study, insulin was covalently attached to two water-soluble polymers containing N-(2 hydroxypropyl)methacrylamide using the mixed anhydride method. The first polymer was a copolymer of N-(2-hydroxypropyl)methacrylamide and N-methacryloyldiglycine. The second one was a terpolymer of two of the above-given monomers and R-(-)-1 methyl-2-methacryloylamidoethyl 2-acetamido-2-deoxy-beta-D-glucopyranoside. Insulin conjugates were isolated and characterized, and the mitogenic effect on SMC was investigated. The results showed that only conjugates of insulin and terpolymers bearing pendant N-acetyl-glucosamine groups do not have a mitogenic effect on SMC while maintaining the hypoglycemic activity of insulin. This finding suggests that some inter- or intramolecular interactions of coupled insulin with the sugar moiety(ies) attached to the polymer backbone contribute to the observed effects. PMID- 8731217 TI - Effects of growth-factor-enhanced culture on a chondrocyte-collagen implant for cartilage repair. AB - The effects of incubation and addition of growth factors to a chondrocyte-seeded collagen implant for cartilage repair were studied. Type I collagen matrices seeded with lapine articular chondrocytes and unseeded controls cultured in the presence and absence of fibroblast growth factor and insulin for 2, 6, and 9 weeks were subjected to biomechanical, biochemical, and histological analysis. Aggregate modulus of elasticity of seeded implants decreased by half at 6 weeks, then rose by a factor of 10 above initial values. Permeability of seeded implants and their controls decreased steadily. Glycosaminoglycan content peaked at 6 weeks, coinciding with the greatest number of chondrocytes and mitotic activity in seeded implants. Chondrocytes remained phenotypically stable and metabolically active; they incorporated glycosaminoglycan into the extracellular matrix, and formed an organized pericellular environment despite the predicted resorption of the collagen matrix. Adding fibroblast growth factor and insulin tripled the rate of cell turnover and doubled the glycosaminoglycan content of seeded implants, but had no effect on their material properties. In vitro incubation for 6 weeks in the presence of fibroblast growth factor and insulin creates a metabolically and mitotically active chondrocyte-collagen composite for implantation into articular cartilage defects. PMID- 8731218 TI - Radiation-induced graft copolymer SBS-g-VP for biomaterial usage. AB - The grafting of 4-vinyl pyridine (VP) to styrene-butadienestyrene triblock copolymer (SBS) by radiation-induced graft copolymerization was studied. The cohesive properties such as cohesive energy (Ecoh), molar volume (V), solubility parameter (delta), molar molecular weight (WM), specific volume (Vg), and density (1/Vg) of SBS-g-VP graft copolymer were calculated according to the group contribution of Fedors. The morphology of SBS-g-VP was studied by optical polarizing microscopy. We also measured the glass transition temperature and the mechanical properties of SBS-g-VP graft copolymer. Contact angle and blood clotting time experiments were also performed to evaluate the biocompatibility of SBS-g-VP. A second domain was found in the SBS-g-VP graft copolymer, which resulted in different properties between SBS-g-VP and SBS. The blood compatibility of SBS-g-VP as measured by the Lee-White clotting test was better than that of SBS and polystyrene. PMID- 8731219 TI - Absorption of iron by dentin: its role in discoloration. AB - This study investigated the effect of iron-containing mordants on the discoloration of human dentin. Dentin wafers with intact enamel borders were treated with one of several acid solutions containing iron. After exposure to aqueous sodium sulfide, color change was evaluated colorimetrically. X-ray photoelectron spectroscopy (XPS) and energy dispersive x-ray spectrometry (EDS) were used to probe surfaces for the presence of iron. Changes in surface morphology were evaluated by scanning electron microscopy (SEM). The results indicated that dentin treated by iron-containing solutions always discolored when the sodium-sulfide concentration exceeded 9.7 mM; variability in discoloration occurred below this level. XPS detected iron in enamel but not in dentin, while EDS detected iron in both enamel and dentin. Since XPS probes the upper atomic layers, these results indicate that acid-demineralized dentin absorbs iron; however, a water wash removes iron from the uppermost dentin surface, suggesting that the iron is physically absorbed. When exposed to sodium sulfide, at least some of the iron remaining in the bulk region is able to migrate to the surface to form iron-sulfide compounds. Based on SEM findings, discoloration is associated with the formation of a surface film on dentin. PMID- 8731220 TI - Progesterone receptor immunoreactivity in aromatic L-amino acid decarboxylase containing neurons of the guinea pig hypothalamus and preoptic area. AB - A double-labeling immunofluorescence procedure was used to determine whether progesterone receptor (PR)-immunoreactive (IR) neurons in the preoptic area and hypothalamus of female guinea pigs also contained aromatic L-amino acid decarboxylase (AADC), an enzyme involved in the synthesis of both catecholamines and serotonin. Immunostaining was performed on cryostat sections prepared from ovariectomized guinea pigs primed by estradiol to induce PR. The nuclear presence of PR was visualized by a red fluorescence while the AADC-containing perikarya showed a yellow-green fluorescence. The topographic distribution of AADC-IR neurons was investigated by using a specific antiserum obtained by immunization of rabbits with a recombinant protein beta-galactosidase-AADC in the two regions known to contain the densest populations of estradiol-induced PR-IR cells: the preoptic area and the mediobasal hypothalamus. The localization of PR-IR and AADC IR cell populations showed considerable overlap in these areas, mainly in the medial and periventricular preoptic nuclei and in the arcuate nucleus. A quantitative analysis of double-labeled cells estimated that about 15% to 23% of AADC-IR cells in the preoptic area and about 11% to 21% of AADC-IR cells in the arcuate nucleus possessed PR. This colocalization persisted throughout the rostrocaudal extent of these areas and represented 3% to 9% of the population of PR-IR cells. These findings provide neuroanatomical evidence that a subset of AADC neurons is directly regulated by progesterone. The exact physiological role of this enzyme in target cells for progesterone is not understood. AADC may be involved in functions other than that for the synthesis of the classical neurotransmitters. PMID- 8731221 TI - Immunohistochemical, electrophysiological, and electron microscopical study of rat fungiform taste buds after regeneration of chorda tympani through the non gustatory lingual nerve. AB - The sensory innervation of fungiform papillae on the rat dorsal tongue is derived from branches of two cranial nerves: the lingual branch of the trigeminal nerve which provides somatosensory innervation and the chorda tympani (CT) branch of the facial nerve, which provides innervation to the taste buds. Removal of the CT results in degeneration of the taste buds. Removal of both nerves results in reduction in size of fungiform papillae and an altered pattern of keratinization in its epithelium. Regeneration of nerves to the epithelium restores the pre operative condition. Thus, in addition to their sensory functions, both the CT and lingual seem to exert trophic effects on the phenotypic expression of epithelial cells in the fungiform papillae. We severed both the CT and lingual nerves in rats and sutured the proximal stump of the CT to the distal stump of the lingual to promote regeneration of the CT along the lingual nerve pathway. At the same time, we prevented the proximal stump of the lingual from regenerating into the tongue. Our purpose was to determine whether and how the innervation pattern of the regenerated taste bud might be different from normal under these experimental conditions. We found that reinnervation by the CT through the lingual nerve occurs, that this restores the anatomical and functional integrity of the fungiform taste buds and papillae, and that some papillae, but not all, were richly innervated with subgemmal, extragemmal, and perigemmal neuron specific enolase, calcitonin gene-related peptide, substance P, and neurokinin A positive fibers. Moreover, responses to taste stimuli were recorded electrophysiologically from the CT. PMID- 8731222 TI - Organization of efferent projections from the spinal cervical enlargement to the medullary subnucleus reticularis dorsalis and the adjacent cuneate nucleus: a PHA L study in the rat. AB - The distribution and organization of projections from the spinal cervical enlargement to subnucleus reticularis dorsalis (SRD) and the neighbouring Cuneate nucleus (Cu) area was studied in the rat by using microinjections of Phaseolus vulgaris leucoagglutinin (PHA-L) into different laminae around the C7 level. The Cu received very dense projections from the dorsal horn, with the highest density being observed following injections into the medial part of laminae III-IV. The SRD received dense projections from laminae V-VII of the cervical enlargement, particularly from the reticular and medial aspects of lamina V, lamina VI, and the dorsal part of lamina VII. By contrast, the superficial part of the dorsal horn (laminae I to IV) and the dorsal part of lamina X provided only sparse projections to the SRD. Clusters of labelled terminals and boutons were observed mainly in the SRD areas subjacent to the Cu. In the caudorostral axis, labelled terminals were spread along the whole SRD from the cervicomedullary junction up to the caudal-most part of the area postrema. Contralateral projections to the SRD were scarce and were observed mainly after injections into the medial part of laminae VI-VII. These data give further support to the proposal that there are two parallel systems in neighbouring structures of the caudal medulla, viz. the Cu and the SRD, which, respectively, relay lemniscal and nociceptive information from the spinal cord to the thalamus. PMID- 8731223 TI - Glutamate metabolic pathways in displaced ganglion cells of the chicken retina. AB - Glutamate (E) is the putative amino acid neurotransmitter used by ganglion cells, photoreceptors, and bipolar cells. Aspartate (D) and glutamine (Q) are potential precursors of glutamate, and glutamate-utilizing neurons may use one or more of these amino acids to sustain production of glutamate. We used post-embedding immunocytochemistry for several amino acid neurotransmitters to characterize the amino acid signatures for displaced ganglion cells of the avian retina. We found two neurochemical signatures for displaced ganglion cells, EQ and EDQ, in mid peripheral and far-peripheral retina, respectively. Differences in neurochemical signatures cannot be explained by the existence of two ganglion cell populations, and we propose that the two signature categories for the large-diameter displaced ganglion cells reflect variations in the aspartate precursor pool. The transamination reaction involved in glutamate production, aspartate/oxaloacetate and alpha-ketoglutarate/glutamate, requires an active TCA cycle, since the carbon skeleton of glutamate is derived from alpha-ketoglutarate, a TCA intermediary. We hypothesized that aspartate levels vary in the normal chicken retina because eccentricity-dependent differences in oxygen availability result in changes of alpha-ketoglutarate levels, and hence, alterations in the equilibrium of the transamination reaction. We tested this hypothesis by incubating isolated chicken retinas in anaerobic conditions and found elevated aspartate immunoreactivity in subpopulations of glutamate-utilizing neurons in the central retina. Under aerobic conditions, or in retinas placed directly into fixative, retinal samples from the central edge of the pecten did not show differential cellular staining for aspartate. We have, therefore, identified differences in neurochemical signatures for retinal neurons involving changes in active maintenance of precursor pools. PMID- 8731224 TI - Morphological classes of spinothalamic lamina I neurons in the cat. AB - We examined the morphology and distribution of retrogradely labeled spinothalamic tract (STT) neurons in lamina I (the marginal zone) of the spinal dorsal horn after large injections of cholera toxin subunit B (CTb) or Fast Blue (FB) into the contralateral thalamus of cats. Based on the shape and orientation of the somata and proximal dendrites in horizontal sections, three distinct cell types were identified: (1) fusiform cells with small, spindle-shaped somata and bipolar, longitudinal dendritic arbors; (2) pyramidal cells with triangular somata and three main dendritic origins with primarily longitudinal arborizations; and (3) multipolar cells with larger, multiangular somata and four or more radiating dendritic arbors directed both longitudinally and mediolaterally. These three morphological types differed significantly in the number of primary dendrites and the size of the somata. Subclasses of multipolar cells were noted. Nearly all cells could be categorized into these three classes consistently in horizontal sections. A small number of cells with transitional shapes or with small, round somata were unclassified. The proportional distributions of these cell types were found to vary over the length of the spinal cord (from the third cervical through the coccygeal segments) in three cats. The overall proportions of cell types were 34% fusiform, 36% pyramidal, 25% multipolar, and 5% unclassified. The proportions of pyramidal and multipolar cells were strikingly higher within the C7-8 and L6-7 segments and lowest in the thoracic segments. In contrast, fusiform cells formed about 20% of the labeled lamina I STT population in the C7-8 and L6-7 segments but more than 60% in thoracic segments. Across all nine cats, the proportions were similar within the cervical (C5-8) and lumbosacral (L5-S1) enlargements, although considerable interanimal variability was noted. These distinct morphological types of lamina I STT cells with differential longitudinal distributions probably have different functional roles. They may correspond to the three main physiological classes of lamina I STT cells. PMID- 8731225 TI - Olfactory glomeruli are innervated by more than one distinct subset of primary sensory olfactory neurons in mice. AB - The rodent olfactory epithelium consists of a mosaic of primary sensory olfactory neurons (PONs) which express distinct putative olfactory receptor proteins. Recent evidence suggests that individual subsets of these sensory neurons project to separate glomeruli in the olfactory bulb (Vassar et al., [1994] Cell 79:981 991). In the present study we have identified two distinct subsets of primary sensory olfactory neurons (PONs) in the H-OMP-LacZ-6 transgenic mouse. In these transgenic mice, a LacZ reporter gene under the control of a 294 base pair element from the 5' promoter region of the olfactory marker protein (OMP) gene was expressed in a subset of PONs located in a discrete band of neuroepithelium in the nasal cavity. These LacZ positive neurons were not randomly located within this band but were more concentrated within a locus between endoturbinates IIb and III. The axons of these neurons densely innervated three adjacent and bilaterally symmetrical glomeruli present in the ventromedial olfactory bulb. Labeling of tissue sections with the plant lectin Dolichos biflorus (DBA) revealed an independent subset of PONs in the transgenic mice. These neurons were present in a wide region of the nasal cavity that included the neuroepithelial band containing the LacZ expressing neurons. The DBA labeled axons terminated in glomeruli in the rostromedial and dorsolateral olfactory bulb surfaces. Although the glomeruli innervated by the LacZ and DBA positive axons were predominantly non-overlapping there were glomeruli in the ventral olfactory bulb that were labeled by both DBA and LacZ markers. Eight different types of glomeruli were characterized. Most notably, glomeruli were identified which were innervated partially by both or by either subset alone. In these cases, axon subsets were observed to terminate within discrete subregions of a glomerulus. These results support the hypothesis that phenotypically distinct subsets of PONs converge on to the same glomeruli but also indicate that some glomeruli are innervated by more than one subset of sensory neuron. These findings have implications for understanding how the olfactory projection is formed and how olfactory information is processed. PMID- 8731226 TI - Tuberous electroreceptor organs form in denervated regenerating skin of a weakly electric fish. AB - Weakly electric fish use tuberous electroreceptor organs to detect their own electric fields. We investigated the role of innervation upon regeneration and differentiation of tuberous electroreceptor organs. The left, infraorbital, anterior lateral line nerve of brown ghosts (Apteronotus leptorhynchus) was sectioned, and the proximal stump was dipped in ricin to prevent regrowth. Immediately after denervation, a piece of cheek skin (approximately 0.5 cm2) was removed bilaterally to induce skin regeneration. After survival periods of 3, 4, or 5 weeks, regenerated skin from the left (denervated) and the right (reinnervated) sides was removed and processed for immunocytochemistry or electron microscopy. Tuberous electroreceptor organs were present in regenerated reinnervated, as well as regenerated denervated skin patches at all survival times. With increased time after skin removal, the number of fully differentiated organs increased in the reinnervated regenerated skin while the number of organs with degenerating receptor cells or entirely devoid of receptor cells increased in the denervated regenerated skin. These results suggest that innervation is not essential for tuberous electroreceptor organ development, but that it is necessary for complete sensory cell differentiation and long-term survival. PMID- 8731227 TI - Light perception in the vertebrate brain: an ultrastructural analysis of opsin- and vasoactive intestinal polypeptide-immunoreactive neurons in iguanid lizards. AB - Recent biochemical and immunocytochemical evidence indicates that a population of circadian and reproductive rhythm-entraining photoreceptors lies in the basal diencephalon of iguanid lizards. Here, we report the results of correlated light and electron microscopy of opsin-immunoreactive cells in the basal brain, and we discuss their ultrastructural relationship to known photoreceptors. Cerebrospinal fluid (CSF)-contacting bipolar neurons in the lizards Anolis carolinensis and Iguana iguana were immunolabeled with antisera generated against vertebrate retinal opsins and vasoactive intestinal polypeptide (VIP). Within the brain, opsin-immunoreactive cells were found exclusively in the ependyma of the basal region of the lateral ventricles (adjacent to nucleus paraolfactorius/nucleus ventromedialis and neostriatum/paleostriatum). Cells in the same anatomical location and with the same morphology were labeled with anti-VIP antisera. These cells possessed a dendritic process that extended toward the lateral ventricle, ending in a bulbous terminal that protruded into the ventricle. Axonal processes travelled ventrally and caudally. The entire cell, including the axonal process, exhibited opsin-like and VIP-like immunoreactivity. By light microscopy, opsin like immunostaining appeared punctate, with immunoreactivity greatest in the bulbous terminal. Opsin- and VIP-immunostained thick sections were resectioned, and individual cells observed by light microscopy were then characterized using electron microscopy. We found that all immunostained cells were morphologically similar and that they were morphologically distinct from neighboring nonimmunoreactive cells. CSF-contacting opsin- and VIP-immunoreactive cells lacked the membranous stacks characteristic of retinal photoreceptors but were ciliated and contained numerous large electron-dense vesicles. Multiple synaptic contacts were made on the soma and putative dendritic processes of opsin- and VIP immunoreactive CSF-contacting neurons. Our results provide the first ultrastructural characterization of opsin-immunostained encephalic CSF-contacting neurons in a vertebrate animal, and they indicate that these putative photoreceptors share structural features with pineal photoreceptors and with certain invertebrate extraretinal photoreceptors, but they are morphologically and biochemically distinct from visual photoreceptors of the retina. PMID- 8731228 TI - GABA plasma membrane transporters, GAT-1 and GAT-3, display different distributions in the rat hippocampus. AB - This study evaluates the distribution of two high affinity gamma-aminobutyric acid (GABA) transporters (GAT-1 and GAT-3) in the rat hippocampus using immunocytochemistry and affinity purified antibodies. GAT-1 immunoreactivity was prominent in punctate structures and axons in all layers of the dentate gyrus. In Ammon's horn, immunoreactive processes were concentrated around the somata of pyramidal cells, particularly at their basal regions. The apical and basal dendritic fields of pyramidal cells also displayed numerous GAT-1 immunoreactive punctate structures and axons. The zone of termination of the mossy fibers that includes both the hilus of the dentate gyrus and stratum lucidum of the CA3 area was the lightest immunolabeled region of the hippocampal complex. Electron microscopic preparations demonstrated that GAT-1 immunoreactive axon terminals form symmetric synapses with somata, axon initial segments, and dendrites of granule and pyramidal cells in the dentate gyrus and Ammon's horn, respectively. Immunoreactivity was localized to the plasma membrane and the cytoplasm of axon terminals. The somata of previously described local circuit neurons in the dentate gyrus and Ammon's horn contained GAT-1 immunoreactivity associated with the Golgi complex. Light, diffuse GAT-3 immunoreactivity was present throughout the hippocampal formation. Thin, astrocytic glial processes displayed GAT-1 and GAT-3 immunoreactivity. This localization of GAT-1 and GAT-3 indicates that they are involved in the uptake of GABA from the extracellular space into GABAergic axon terminals and astrocytes. PMID- 8731229 TI - NADPH-diaphorase localization in the CNS and peripheral tissues of the predatory sea-slug Pleurobranchaea californica. AB - The distribution of putative nitric oxide synthase (NOS)-containing cells in the opisthobranch mollusc Pleurobranchaea californica was studied histochemically via NADPH-diaphorase (NADPH-d) reduction of Nitro Blue Tetrazolium (NTB). Whole mounts and cryostat sections were prepared from the central nervous system and peripheral organs, including the buccal muscles, esophagus, salivary glands, foot, mantle, and gills. NADPH-d-positive neurons were localized predominantly to the buccal and pedal ganglia as well as to distinct areas of the cerebropleural and visceral ganglia. A variety of identified neurons were positive for NADPH diaphorase in various central ganglia, including the metacerebral cells of the cerebropleural ganglion, putative locomotor neurons of the pedal ganglia, and buccal motoneurons. Specific staining was observed only in somata of central neurons, whereas neuropil areas remained unstained. However, NADPH-d-reactive axons were dense in buccal ganglion nerves, whereas peripheral nerves and connectives of other ganglia had few or no NADPH-d positive terminals. In the periphery, NADPH-d activity was detected only in a few neurons of the rhinophore and tentacle ganglia. NADPH-d staining was marked in the salivary glands and gills, but there was no or very little staining in the esophagus, buccal mass, and foot. Histochemical stain production required the presence of both beta-NADPH and NBT; alpha-NADPH could not substitute for beta-NADPH. The inhibitor of NOS, 2,6-dichlorophenol-indophenol, at 10(-3) M, totally abolished NADPH-d-positive staining. The apparent high activity of central NADPH-d contrasts with much lower activity in the ganglia of the related gastropod Tritonia. These data suggest a role for nitric oxide as a signal molecule in the central nervous system of Pleurobranchaea. PMID- 8731230 TI - Mitotic neuroblasts determine neuritic patterning of progeny. AB - Neuronal precursor proliferation and axodendritic outgrowth have been regarded as strictly sequential, with process formation presumably beginning after mitotic activity ceases. We now report that sympathetic precursors in vitro often elaborate long neurites before dividing. Of 437 neuroblasts observed in 48 time lapse recordings, 42 neuroblasts divided. Thirty (71%) of these mitotic neuroblasts had neurites prior to cytokinesis. "Paramitotic" neurites were found to contain microtubules (MTs), indicating that precursors elaborate neuritic cytoskeleton during proliferation. Remarkably, the precise neuritic pattern exhibited by parental neuroblasts was consistently reproduced by daughter cell pairs. Preservation of neuritic morphology occurred through asymmetric division, with individual neurites allocated to specific daughter cells. Paramitotic neurites either remained intact throughout mitosis (12 of 65), or "retracted" into the soma during prophase and then "regrew" within minutes after cytokinesis (53 of 65). "Retraction" and "regrowth" involved resorption of cytoplasm into the soma, then refilling of residual cell membrane, resulting in recapitulation of the parental neurite pattern. Paramitotic neuritogenesis appears to be intrinsically driven, but is responsive to environmental signals. The culture substrate influenced neurite length, but not the response of paramitotic neurites during mitosis or the preservation of neuritic morphology. However, the incidence of neurite-bearing neuroblasts increased from 38 +/- 1.3% to 94 +/- 1.1% with growth factor treatment. The surprisingly high incidence of paramitotic neurites and the fidelity with which patterning was conserved across cell generations raise the possibility that mitotic precursors engage in pathfinding. Our studies suggest a novel link between neurogenesis and cytoarchitectonic patterning. PMID- 8731231 TI - Cytokines in juvenile arthritis. PMID- 8731232 TI - From classification criteria to diagnostic criteria for rheumatoid arthritis. PMID- 8731233 TI - Dynamics of fever and the cytokine network in systemic juvenile arthritis. AB - AIM OF THE STUDY: To determine the levels of the inflammatory cytokines IL1 alpha, IL1 beta, TNF alpha, IL6, IL8 and of their inhibitors TNF-sR55, TNF-s R75 and IL1-Ra during temperature elevation in systemic juvenile chronic arthritis. METHODS: Fifty-six serum samples were collected at regular intervals from seven children during 8 fever cycles. Cytokine levels were determined using enzyme linked immunoassays. RESULTS: Levels of IL1 alpha, IL1 beta, TNF alpha and IL8 showed no variations. In contrast, IL6 and IL1-Ra levels paralleled the fever spikes. TNF-sR75 levels were also correlated with the fever. CONCLUSION: Fever dynamics in systemic juvenile chronic arthritis may be partly related to cytokine variations. PMID- 8731234 TI - Interleukin-6 and soluble interleukin-2 receptor in juvenile chronic arthritis: correlations with clinical and laboratory parameters. AB - We sought to determine whether levels of interleukin-6 and soluble interleukin-2 receptor were correlated with clinical parameters including functional capacity indices such as Steinbrocker's class and the Juvenile Arthritis Functional Assessment Report (JAFAR) score, with tests for inflammation, and/or with immunological parameters in 24 patients with active polyarticular or pauciarticular juvenile chronic arthritis. Levels of interleukin-6 and soluble interleukin-2 receptor were significantly higher in juvenile chronic arthritis patients than in healthy controls (p < 0.005 and p < 0.00005, respectively). Interleukin-6 levels were correlated with the following parameters: number of painful joints (p < 0.025); Ritchie's index (p < 0.025); visual analog scale pain score (p < 0.025); Steinbrocker's class (p < 0.025); JAFAR score determined by patients (p < 0.05); JAFAR score determined by parents (p < 0.05); erythrocyte sedimentation rate (p < 0.0002); and serum levels of C-reactive protein (p < 0.0003), hemoglobin (p < 0.05), albumin (p < 0.025), and alpha 2-globulins (p < 0.025). Levels of soluble interleukin-2 receptor did not correlate with any of the parameters studied. Levels of interleukin-6 and soluble interleukin-2 receptor were not correlated with each other. Abnormal levels of interleukin-6 or soluble interleukin-2 receptor were not significantly associated with the presence of antinuclear antibodies, IgM-rheumatoid factor, IgA rheumatoid factor or anticardiolipin antibodies. Our findings suggest that interleukin-6 is a useful parameter for assessing juvenile chronic arthritis and that the potential clinical value of elevated levels of soluble interleukin-2 receptor in this disease needs to be further evaluated in longitudinal studies. PMID- 8731235 TI - Influence of severity of chronic inflammatory joint disease on the pharmacokinetics of indomethacin and etodolac. AB - The goal of this study was to look for correlations between the severity of chronic inflammatory joint disease and pharmacokinetic parameters of nonsteroidal antiinflammatory drugs. Disease severity data (pain severity and magnitude of abnormalities in laboratory tests for inflammation) and pharmacokinetic data (area under the curve in the morning (AUCm) and maximum plasma concentration (Cmax) were collected during a prospective, randomized, double-blind, parallel group study. Two groups of nine and 11 patients, respectively, were given 300 mg etodolac b.i.d or 50 mg indomethacin b.i.d. by the oral route, for three days, after a 36-hour placebo washout. Univariate analyses demonstrated statistically significant negative correlations between pharmacokinetic parameters of both study drugs and a number of disease severity parameters. In the multivariate analysis of data for etodolac, the sigma erythrocyte sedimentation rate contributed significantly to variations in all pharmacokinetic parameters and explained 100% of the variations in free S-enantiomer AUCm and in total and free S-enantiomer Cmax. For indomethacin, pain contributed to variations in Cmax values of the total and free forms; the sigma erythrocyte sedimentation rate was also a factor in variations in total indomethacin. These negative correlations suggest that severity of chronic inflammatory joint disease may influence the pharmacokinetics of nonsteroidal antiinflammatory drugs. PMID- 8731236 TI - How do French rheumatologists treat early rheumatoid arthritis? AB - Although factors that appear to predict long-term outcomes of rheumatoid arthritis have been identified, there is no consensus about the treatment early in the disease. To determine how French office- and hospital-based rheumatologists treat early rheumatoid arthritis, we created three clinical vignettes corresponding to different levels of severity of early rheumatoid arthritis (less than six months' disease duration). Cases 1 and 2 were relatively young patients (35 and 50 years), and Case 1 had numerous poor prognosis factors. Case 3 was 80 years of age. Rheumatologists were asked to indicate which medications they would use at presentation and after one year of a favorable or unfavorable course. The study was conducted by questionnaire (response rate, 58%). Of the 185 rheumatologists who completed the questionnaire, 81% were male and 19% female; mean age was 42 +/- 8 years. In Cases 1 and 2, nonsteroidal antiinflammatory drugs were given by 99% of respondents; second-line drugs were prescribed at presentation by 93% of respondents in Case 1 and 86% in Case 2, and methotrexate was more likely to be used in the presence of poor prognosis factors (23% in case 1 and 7% in Case 2). In the event of an unfavorable course after one year, a larger proportion of rheumatologists prescribed glucocorticoid therapy (65% in Case 1 and 20% in Case 2), and there was a shift from "conventional" to "modern" second-line drugs, with more widespread use of methotrexate (65% in case 1 and 18% in case 2). In the 80-year-old patient, glucocorticoid therapy was used more often than nonsteroidal antiinflammatory drugs and second-line drugs (gold salts, hydroxychloroquine, sulfasalazine) were prescribed by 40% of rheumatologists at presentation and by 67% after one year of an unfavorable course; in the latter situation, methotrexate was selected in 24% of cases. In contrast to conventional recommendations, many French office- or hospital-based rheumatologists use second-line drugs very early and base their choice of medications on the estimated risk of severe disease and on the age of the patient. PMID- 8731237 TI - Results of surgery for lumbar spinal stenosis in patients aged 80 years or more. A retrospective study of thirty-four cases. AB - As life expectancy increases and spinal imaging techniques improve, surgery is being increasingly viewed as a therapeutic alternative for symptomatic lumbar spinal stenosis in patients older than 80 years. Thirty-four patients (21 men and 13 women) who had surgery for lumbar spinal stenosis in our department between 1979 and 1994 were studied retrospectively. The most common initial symptoms were walking-related disorders (n = 29) and sciatica or femoral neuralgia (n = 34). All 34 patients underwent laminectomy at one or more levels. Ten patients also had a herniated disk. There were no deaths and only two patients had serious complications (persistent foot drop in one and left-sided hemiplegia in the other). Results were evaluated immediately after surgery and after three and 12 months. The overall result on pain and walking-related disorders was good in 53% of cases, acceptable in 32%, and poor in 15%. Our data suggest that surgery is a reasonable alternative in symptomatic elderly patients who are in good general health. Satisfactory results can be obtained although disabling complications can occur. PMID- 8731238 TI - Osteoporotic fractures revealing anorexia nervosa in five females. AB - Anorexia nervosa affects 0.5% to 1% of female adolescents. The course is chronic in 50% of cases, causing substantial bone loss with osteoporotic fractures after a few years of amenorrhea. This is probably an underestimated problem. The diagnosis of anorexia nervosa is readily missed, as illustrated by five cases reported herein. The five patients were females aged 17 to 44 years who were only slightly underweight (mean weight, 43.6 kg; body mass index < 20 kg/m2). The first fracture occurred seven to 24 years after the onset of anorexia nervosa. Three patients had vertebral crush fractures, and two had peripheral insufficiency fractures. Bone mineral density measured by absorptiometry was very low (mean lumbar z-score, -3.7 SD). Three patients, who were all members of health care professions, knew that they had anorexia nervosa but failed to report this condition. In the other two patients, the amenorrhea had been mistakenly ascribed to other causes (Stein-Leventhal syndrome and psychogenic anovulation). None of the patients was receiving medical follow-up. Anorexia nervosa should be considered routinely in women who are slightly underweight. The patients often deny abnormal menstruation or eating behaviors. The diagnosis rests on determination of the body mass index, a thorough history emphasizing current and past gonadal dysfunction, and evaluation of the diagnostic criteria for anorexia nervosa. Osteoporosis is probably a common but underestimated complication of anorexia nervosa, particularly before the menopause. Enhanced awareness of this condition should allow earlier detection of a greater number of cases. PMID- 8731239 TI - Epidural lipomatosis not induced by corticosteroid therapy. Three cases including one in a patient with primary Cushing's disease (review of the literature). AB - We report three cases of epidural lipomatosis including one in a patient with primary Cushing's disease. Our literature review found 16 additional cases of symptomatic epidural lipomatosis in patients who were not receiving corticosteroids. The presenting symptoms were nonspecific. The main clinical symptoms were nerve root pain, weakness of the lower limbs upon exertion, paraparesis or isolated back pain. Degenerative lesions were common and were sometimes the cause of the symptoms. Cases were evenly distributed between the thoracic and lumbar spine. Of the 18 patients, 14 were men and eight were older than 54 years. Three-fourths of patients were obese. Spinal cord or nerve root compression occurred in some instances. Modern imaging techniques (computed tomography and magnetic resonance imaging) can establish the diagnosis rapidly. In patients without neurologic compromise, surgery should be considered only if symptoms fail to respond to weight reduction. The rate of occurrence of epidural lipomatosis in patients with Cushing's disease is probably underestimated. Routine investigation by magnetic resonance imaging of Cushing's disease patients who have manifestations known to occur in epidural lipomatosis would allow to evaluate the role of increased production of endogenous corticosteroids in the occurrence of epidural lipomatosis. PMID- 8731240 TI - Cystic lung disease in systemic sclerosis: a case report with high resolution computed tomography findings. AB - A 31-year old black female presented with limited scleroderma. Although she had no chest symptoms, chest radiographs showed a basal reticular pattern with large cysts in both lung fields. High resolution computed tomography revealed almost complete effacement of normal lung architecture by multiple large cysts, honeycombing and interstitial fibrosis. Cystic lung disease is an uncommon manifestation of interstitial lung disease in systemic sclerosis, and its natural history needs to be better defined. PMID- 8731241 TI - Reactive arthritis induced by Strongyloides stercoralis. AB - Reactive arthritis induced by Strongyloides is exceedingly rare. A case in a 53 year-old man from the Guadeloupe (French Antilles) is reported. The outcome was rapidly favorable under thiabendazole therapy. The cycle of Strongyloides is reviewed, and the contribution of parasites to reactive arthritis in patients with genetic risk factors is discussed. Establishing the correct diagnosis is sometimes difficult but is essential in order to avoid inappropriate administration of corticosteroids that can lead to fatal, multivisceral dissemination of the parasite, particularly in patients with strongyloidiasis. PMID- 8731242 TI - Myositis ossificans circumscripta with spontaneous healing of a cortical cyst. AB - We report a case of myositis ossificans circumscripta with an initial periosteal reaction and subsequent development of a cyst in the underlying cortex. All the roentgenographic abnormalities resolved spontaneously and permanently. The unusual course may have been due to formation of a subperiosteal hematoma at the time of development of the soft tissue ossification. PMID- 8731243 TI - Hypersensitivity to glucocorticoids: does it exist? AB - Hypersensitivity to corticosteroids is a classical but rarely reported event. We report a 30-year-old patient who developed generalized urticaria after her first methylprednisolone bolus. We reviewed the relevant literature to look for factors associated with hypersensitivity to corticosteroids. Causality should be evaluated on a case-by-case basis using diagnostic criteria for drug hypersensitivity reaction. Etiopathogenesis may involve either an IgE-mediated immunoallergic reaction or semi-delayed hypersensitivity. The main problems are identification of the offending agent and evaluation of the safety of further corticosteroid therapy. Although a few fatal reactions have been reported, some were probably due to underlying cardiovascular disease or serum electrolyte abnormalities. PMID- 8731244 TI - Antiphospholipid antibodies and movement disorders. PMID- 8731245 TI - Acute monoarthritis in a patient under isotretinoin. PMID- 8731246 TI - Tuberculin-induced rheumatic manifestations. A new case. PMID- 8731247 TI - [An unprotected flank in the immunologic prevention of tetanus]. PMID- 8731248 TI - [Colectomy and anterior resection with a video-laparoscopic approach. Initial experience-results]. AB - Laparoscopic procedures are rapidly advancing. Laparoendoscopic colectomy appears to be an option the treatment of colonic disease and is increasingly being performed in selected patients. The purpose of this study is to review our initial experience with the method which initiated in 1992. Thirty-three patients underwent laparoscopic resection of colorectal segment. All the patient, charts were assessed and data were obtained about the diagnosis, operation performed, complications, conversion and postoperative course. Diverticular disease was the most frequent (54.5%) indication of laparoscopic colorectal procedure in our series followed by adenocarcinoma (27.3%). Left colectomy was performed in 19 (57.6%) patients. Anastomosis followed double-stapling technique in most of them. Conversion to open procedure occurred in six (18.2%) cases. There were 4 complications associated to the method resulting in an specific morbidity of 12.1% in this series. There were no complications related to the anastomosis and no death occurred. Postoperative course was favorable for the patients and all could take oral liquids in the day after the procedure. Colorectal surgery performed by video-laparoscopic access is feasible, safe and benefits all patients. Conversion and morbidity in our series are similar to those shown in literature. It must, however, be indicated in a selected basis and in centers of reference for treatment of colorectal disease. PMID- 8731249 TI - [Lung lesion in acute pancreatitis]. AB - Clinically detectable signs of lung injury develop in up to 70 percent of patients with acute pancreatitis. In order to study the pulmonary injury, experimental haemorrhagic pancreatitis was induced in 63 Wistar rats by intraductal injection of 5 per cent sodium taurocholate. Investigations were carried out 2, 4, 6, 12, 24 and 48 hours after the end of pancreatitis induction. Lung injury was maximal at 12 hours after pancreatitis induction, pancreatic enzymes (amylase and trypsin) in peritonial fluid were maximal early (2-4h) and serum levels were maximal at about 4 hours after induction. In conclusion, in experimental acute pancreatitis pulmonary injury occurs 12 h after the start of infusion with increase in vascular permeability of the lung. This lesion may be related to pancreatic enzymes in peritonial fluid and blood. PMID- 8731251 TI - [Inguinal hernia repair with the Shouldice technique]. AB - The authors analyze the result of Shouldice inguinal hernia repair in 1000 patients. In a total of 1034 hernias, 966 of them were unilateral and 68 bilateral. The hernias were indirect (61.7%), direct (24.9%) and combined (3.5%). There were 102 recurrent hernias (9.9%) and the mean age of the patients was 50 years. In most cases the patients were male (96.9%) and peasants (47.6%). The post-operative follow-up was annual in 65.8% of the patients, for a period up to 10 years. The recurrence rate was 0.8% in the cases of primary hernias and 4.9% in the case of recurrent ones. PMID- 8731250 TI - [Main variations of the extrahepatic biliary system and their application to the so-called "split-liver" transplantation technique]. AB - Sixty fresh adult livers were obtained from cadavers together with celiac trunk, head of the pancreas and superior mesenteric artery. We have found a single right hepatic duct in 13(21.6%) of the cases and a multiple right hepatic duct in 47(78.3%). A single left hepatic duct was found in 3(5%) of the cases and a multiple left hepatic duct in 57(95%). A median hepatic duct was found in 18(30%) of the cases, out of this 2(3.3%) was multiple median hepatic duct. PMID- 8731252 TI - [Pyoarthritis of the knee--caseload of the Hospital das Clinicas da FMUSP 1990 1994 (86 cases)]. AB - The authors performed a prospective study of 86 patients with septic arthritis of the knee between 1990 e 1994. The age, sex, predisposing factors, time between onset of symptoms and treatment, etiologic organism and their route of entrance in the joint, previous use of antibiotic the data from of treatment, stay at the hospital and complications were evaluated and compared with the literature. PMID- 8731253 TI - Gastric emptying of 99mTc tagged solid food in gastric and duodenal ulceration. AB - This study is to analyze gastric emptying (GE) of 99mTc tagged solid meals in 43 gastric ulcer (GU), 16 duodenal ulcer (DU) patients, and 15 normal subjects. Diagnosis of active ulcers had been made through high gastrointestinal endoscopy. The patients with GU were divided into three groups according to Johnson's classification. Gastric contents were measured at 15 minute intervals over a period of 150 minutes, with the person standing in front of a gamma-counter. The GE curve was obtained from the mean percentile of radioactivity of all the groups measured at 15 minute intervals. Results were submitted to statistical analysis. Significant delay in GE in DU patients compared to normals, type II and Type III group. However, GE of type I GU was not statistically different from the DU group nor of types II and III GU, occupying an intermediate e position. PMID- 8731254 TI - Dyslipidaemias in white normotensive subjects and in white patients treated for essential hypertension. AB - Serum lipid profiles were evaluated in 143 white patients treated for essential hypertension (HT:31 men, 112 women, mean age 58.4 +/- 0.9 years) and in 54 normotensives (NT: 15 men, 39 women, 56.7 +/- 1.8 years). Mean values of LDL-C, HDL-C, TG and LDL-C/HDL-C ratio in hypertensives and normotensives were respectively 4.19 +/- 0.08 vs 4.01 +/- 0.16 mmol/l; 1.14 +/- 0.03 vs 1.22 +/- 0.05 mmol/l, 1.80 +/- 0.10 vs 1.30 +/- 0.06 mmol/l, and 4.0 +/- 0.1 vs 3.5 +/- 0.2; TG and ratio values of both groups differed significantly. Normotensives and hypertensives showed the same prevalence of IIa dyslipidaemia phenotype (37%). Hypertriglyceridaemic phenotypes and associated low HDL-C were more frequent in hypertensives than in normotensives (respectively 20% vs 6% and 14% vs 7%). The IIa phenotype was more prevalent in normotensive women than in men (44% vs 20%). This gender difference was absent in hypertensives (men, 39% vs women, 37%). Isolated low HDL-C was more prevalent in men than in women: NT, 20% vs 8%; HT, 16% vs 4%. IIa prevalence was greater in the age group > 50 years than in the age group < or = 50 years: NT, 22% vs 44%; HT, 12% vs 46%. The prevalence of hypertriglyceridaemic dyslipidaemias was similar in both age groups, in hypertensives (18% vs 21%) and normotensives (6% vs 6%). IIa, IIb and IV dyslipidaemias were more frequent in obese hypertensives. Positive correlations between BMI vs LDL-C and TG values were found in normotensives. In conclusion, the prevalence of dyslipidaemias was similarly high in both NT and HT groups (54% vs 64%), with some qualitative differences concerning phenotypes. PMID- 8731255 TI - [Iron-deficiency anemia in enteral nutrition: correlation with tube position]. AB - Enteral alimentation is a commonly employed procedure for pre-operative rehabilitation and nutritional maintenance of depleted patients suffering from esophageal diseases. In a prospective study of 61 cases exhibiting benign (n = 38) and malignant (n = 23) conditions, a commercial polymeric diet was supplied in the proportion of 40 Kcal/kg/day during at least two weeks. The liquid preparation was infused by bolus technique at home, by the patients themselves, via nasogastric (n = 25) or nasoduodenal silastic tube (n = 36). Total food intake, that before treatment had been under 60% of estimated requirements, reached the expected range throughout the therapeutic period, and also iron input was adequate (0.3 mg/kg/day). Standart anthropometric and biochemical assessment of these cases, with included serum iron and hemoglobin concentration, indicated stabilization of general nutritional parameters, but with persistence of moderate anemia(hemoglobin 12.6 +/- 2.0 g/dl; iron 59.2 +/- 35.7 mg/dl). Stratification of the population according to enteral tube position demonstrated similar diagnoses and clinical conditions for both groups. Mean initial iron and hemoglobin concentration was also the same, but final serum iron was increased after gastric administration (p < 0.05) and diminished with duodenal diet (p < 0.05). It is concluded that: home enteral nutrition in malnourished patients with esophageal diseases is consistent with an acceptable nutritional status, but without significant recovery from anemia, serum iron in this population only increases after intra-gastric alimentation; it is probable that this last finding is related to impaired ionization and absorption of dietary iron, when food is introduced in a postpyloric site. PMID- 8731256 TI - [Cardiovascular effects of arginine and nitric oxide]. AB - Amino acids are an unusual chemical category because they are endowed with alimentary properties, but at the same time they may determine typical pharmacologic responses in some circumstances. These facts were responsible for the origin of modern pharmaconutrition, which has been using arginine for some time because of its beneficial impact on immune function, nitrogen anabolism and wound healing. The recent identification of its metabolite NO (nitric oxide), a powerful mediator related to a large number of physiologic and pathophysiologic phenomenons, has augmented the interest for this amino acid. PMID- 8731257 TI - [Research and teaching in medical education]. AB - A list of citations of scientific papers of Brazilian researchers published from 1981 to 1993 and prepared by Institute for Scientific Information identified almost 8000 titles; 37.4% of this total number include medical area subjects publications. The author analyses also the relevant importance of teaching activities in medical education process, specially with reference to ethical aspects included in it. PMID- 8731259 TI - Molecular typing of Candida albicans strains isolated from nosocomial candidemia. AB - Yeasts of the genus Candida have been recognized as important microorganisms responsible for nosocomial fungemia. Six blood-stream and two intravenous central catheter C. albicans strains were isolated from eight patients and studied by electrophoretic karyotyping of chromosomal DNA by pulsed-field gel electrophoresis. Seven chromosomal DNA profiles were identified. Two patients showed isolates with the same profile, suggesting nosocomial transmission. Karyotyping of C. albicans revealed an excellent discriminatory power among the isolates and may therefore be useful in the study of nosocomial candidemia. PMID- 8731258 TI - Leishmania (viannia) panamensis-induced cutaneous leishmaniasis in susceptible and resistant mouse strains. AB - We studied the susceptibility to Leishmania (Viannia) panamensis in strains of mice. The C57BL/6 strain was resistant and showed self-controlled lesion at the injected foot pad. The BALB/c and DBA/2J strains were susceptible and showed a foot swelling that started day 20 post-infection and progressed to a tumour-like lesion in later period of observation. The CBA/HJ strain was found to be of intermediary resistance. In contrast to other known cutaneous leishmaniasis in mice, the lesion in L. (V.) panamensis-infected mice was restricted to the inoculation site in the skin. In addition, we studied the development of cellular response and antibodies against Leishmania antigen in BALB/c and C57BL/6 strains. The proliferative response of lymph node cells against L. (V.) panamensis antigen was biphasic in both strains. An initial response was seen on day 20, followed by a refractory period between 40 and 80 days and a second response around fourth month post-infection. The response in the latter period was higher in C57BL/6 strain than in BALB/c strain. BALB/c strain presented much higher anti-Leishmania antibody level than C57BL/6 strain. The model and the correlation of immunological variables and the course of the infection are discussed. PMID- 8731260 TI - Association of human herpesvirus 6 infection with exanthem subitum in Belem, Brazil. AB - Recent human herpesvirus 6 (HHV-6) infection was detected in cases of exanthem subitum (ES) involving four children, aged 10 to 24 months, between April and August 1994, in Belem, Brazil. By using the indirect immunofluorescence antibody assay (IFA), significant increases (at least eight times) in antibody concentrations were noted from the acute to the convalescent serum samples, with titers ranging from < 1:10/1:80 to < 1:10/1:640 (patients 3 and 2, respectively). All children had high fever (over 39 degrees C) for three days, followed by generalized, maculo-papular skin rash. A physical examination of the children also revealed concomitant, cervical lymph node swelling and tonsillar pharyngitis in two of them. PMID- 8731261 TI - Effect of skin colour and selected physical characteristics on Schistosoma mansoni dependent morbidity. AB - The effect of the colour group on the morbidity due to Schistosoma mansoni was examined in two endemic areas situated in the State of Minas Gerais, Brazil. Of the 2773 eligible inhabitants, 1971 (71.1%) participated in the study: 545 (27.6%) were classified as white, 719 (36.5%) as intermediate and 707 (35.9%) as black. For each colour group, signs and symptoms of individuals who eliminated S.mansoni eggs (cases) were compared to those who did not present eggs in the faeces (controls). The odds ratios were adjusted by age, gender, previous treatment for schistosomiasis, endemic area and quality of the household. There was no evidence of a modifier effect of colour on diarrhea, bloody faeces or abdominal pain. A modifier effect of colour on hepatomegaly was evident among those heaviest infected (> or = 400 epg): the adjusted odds ratios for palpable liver at the middle clavicular and the middle sternal lines were smaller among blacks (5.4 and 6.5, respectively) and higher among whites (10.6 and 12.9) and intermediates (10.4 and 10.1, respectively). These results point out the existence of some degree of protection against hepatomegaly among blacks heaviest infected in the studied areas. PMID- 8731262 TI - Occurrence of Giardia lamblia in children of municipal day-care centers from Botucatu, Sao Paulo State, Brazil. AB - Considering that the number of day-care centers for pre-school-age children has expanded rapidly in developing countries, and that these institutions presenting conditions that facilitate the transmission of many enteric agents, a parasitological survey was carried out in three municipal day-cares from Botucatu: two in the urban area (one in downtown area and the other one in the city periphery area) and the third in the rural area. Three separate stool specimens were collected from 147 children ranging from 0 to 72 months old and 20 staff members. Each stool specimen was processed by Lutz and zinc sulfate flotation methods. The frequency of giardiasis observed among children of downtown, periphery and rural day-cares was 69.6%, 52.7% and 69.6%, respectively. Only one employee was positive for G. lamblia. The examination of three stool specimens increased the positivity for G. lamblia: from the ninety three final positive examinations, 24 (25.5%) and 8 (8.5%) were positives only after examination of the second and third samples, respectively. Others intestinal organisms like Ascaris lumbricoides (20.4%), Trichuris trichiura (19.0%). Hymenolepis nana (8.8%), Entamoeba coli (22.4%) and Blastocystis hominis (32.0%) were frequently found in the children. There was no significant association among localization of the day-cares, sex of the children and the levels of G. lamblia infection. According to the age, G. lamblia was found mainly in children between 12 to 47 months old. PMID- 8731263 TI - Dengue type 2 outbreak in the south of the state of Bahia, Brazil: laboratorial and epidemiological studies. AB - During March 1994 cases of a exanthematic acute disease were reported in the municipalities of Itagemirim, Eunapolis and Belmonte, state of Bahia. Dengue fever was confirmed by serology (MAC-ELISA) and by dengue virus type 2 isolation, genotype Jamaica. Signs and symptoms of classic dengue fever were observed with a high percentual of rash (73.8%) and pruritus (50.5%). Major haemorrhagic manifestations were unfrequent and only bleeding gum was reported. Dengue virus activity spreaded rapidly to important tourism counties like Porto Seguro, Ilheus, Santa Cruz de Cabralia, Prado, Alcobaca and others, representing a risk for the spreading of dengue virus into the country and abroad. PMID- 8731264 TI - Plague surveillance in Brazil: 1983-1992. AB - Plague caused by Yersinia pestis, has persisted in Brazil in several natural foci spread throughout rural areas in the States of Ceara, Paraiba, Pernambuco, Piaui, Rio Grande do Norte, Alagoas, Bahia, Minas Gerais and Rio de Janeiro. Nationwide surveillance of plague in Brazil based on serological testing started in 1983. We now present an update report of the examinations carried out in our laboratory from 1983 to 1992. The passive hemagglutination test for antibodies against fraction 1A antigen of Y. pestis and the passive hemagglutination inhibition control were employed for testing a total of 220,769 sera. Samples analyzed included 2,856 sera from clinically diagnosed plague cases or suspects, 49,848 sera from rodents of 24 species and 2 species of small wild carnivores (marsupials), 122,890 sera from dogs, and 45,175 sera from cats. Specific antibodies were found in 92 (3.22%) human sera; 143 (0.29%) sera from rodents of 8 species and from the two species of marsupials, 1,105 (0.90%) sera from dogs and 290 (0.64%) sera from cats. The presence of significant levels of specific anti-F1A antibodies among rodents and wild or domestic carnivores (dogs and cats) indicates that all the Brazilian plague foci remain active in spite of the absence of human cases in some of them. PMID- 8731265 TI - Secondary dengue infection in schoolchildren in a dengue endemic area in the state of Rio de Janeiro, Brazil. AB - A seroepidemiologic survey was carried out in schoolchildren from public schools of the Niteroi municipality, state of Rio de Janeiro, Brazil, after a period of sequential epidemics by dengue virus type 1 and 2 (DEN-1 and DEN-2). 450 blood samples were obtained by fingertip puncture and collected on filter paper discs. The hemagglutination inhibition (HAI) test was carried out using DEN-1 and DEN-2 antigens. HAI titres were demonstrated in 66% (297/450) of the sera and the geometric means of the titres were 1/182 and 1/71 for DEN-1 and DEN-2, respectively. Secondary infections were observed in 61% (181/297) of positive cases. Among these, 75% (135/181) were under fifteen years old. No dengue haemorrhagic fever (DHF) was reported in these children. Asymptomatic or oligosymptomatic infections were detected in 56% of the studied population. The absolute and relative frequencies of positive tests by age group and sex did not evidence statistically significant difference. The number of individuals infected probably produced a immunologic barrier responsible for the non occurrence of dengue epidemic in the latter years. PMID- 8731266 TI - Human pulmonary dirofilariasis: a review. AB - The authors presented a detailed summary of the geographical distribution, clinical and pathological aspects of human pulmonary dirofilariasis. Although benign, this zoonosis, of which Dirofilaria immitis is the major etiological agent, represents a medical problem since it produces symptoms which may be confused with neoplasia and thus may subject patients to unnecessary thoracic surgery. Of 229 cases cited in the literature, only 17 were reported in Brazil, despite the existence of highly favorable conditions for the transmission of this infection in man. Thus it may well be that this parasitic infection remains underdiagnosed. Finally, the importance of a differential diagnosis between dirofilariasis and pulmonary neoplasia is emphasized in cases where there is a solitary subpleural nodule ("coin lesion") present. In addition, the development and improvement of modern immunological diagnostic techniques are essential to distinguish this benign disease from other pathological conditions and thus avoid unnecessary surgery. These techniques may reveal the true prevalence of this parasitic infection in our environment. PMID- 8731267 TI - Histoplasmosis in the Muniz Hospital of Buenos Aires. AB - Some epidemiological and immunological characteristics and the methodology of diagnosis of 44 cases of histoplasmosis (HP); 36 (27 males and 9 women) associated with AIDS (HP+AIDS) and 8 (7 males and 1 female) with other predisposing factors (HP+non AIDS), diagnosed in the Muniz Hospital (MH) during 1994, were retrospectively studied. The median age (MA) of HP+AIDS patients was 28 years; 25.5 (22-40) in the women and 28.5 (20-42) in the men and 50 (22-58) years in the HP+non AIDS patients. The more frequent risk factors for HIV infection were intravenous drug addiction (55%) and homo/bisexuality (19%). The MA of these groups were 28 (20-39) and 41 (26-42) years, respectively. Tobaccoism was a predisposing factor in 83% of HP+non AIDS patients. The muco-cutaneous lesions scraping and blood-cultures established the initial diagnosis in 53% and 36% of HP+AIDS patients, respectively and the muco-cutaneous lesions biopsies in 75% of HP+non AIDS cases. At time of diagnosis, all HP+AIDS patients had < 200 while HP+non AIDS patients had > 200 CD4 + lymphocytes/microliters. Seventy two per cent of HP+AIDS patients were born in Buenos Aires (Bs As) city and 62% of HP+non AIDS patients were born in provinces of Argentina other than Bs As. At moment of diagnosis, 87.5% of HP+AIDS and 62.5% of HP+non AIDS patients lived in Bs As city and Bs As outskirts. PMID- 8731268 TI - Carriage of the classical thermotolerant Campylobacters in healthy domestic animals from eastern Peru. PMID- 8731269 TI - Blood culture: specificity in childhood bacterial pneumonia. PMID- 8731270 TI - Sabin-related poliovirus vaccine strains isolated from transverse myelitis cases in Brazil. PMID- 8731271 TI - Acute primary cutaneous Nocardia asteroides infection in a patient with systemic lupus erythematosus. Case report. AB - We report a case of acute primary cutaneous infection of traumatic origin caused by Nocardia asteroides, appeared as cellulitis in a patient with systemic lupus erythematosus. Diagnosis was established by direct examination and cultures from aspirate specimens. The clinical forms of Nocardia infections that affect the skin, reported in Rio Grande do Sul and Uruguay, are discussed. PMID- 8731272 TI - [Determination of the trauma severity level]. AB - The severity of traumas in hospitalized patients was characterized by means of the Injury Severity Score (ISS) was studied prospectively. One hundred trauma patients hospitalized in a trauma referral health facility located in S. Paulo, Brazil, were analyzed. Of the total number of patients, 68 had blunt trauma and 32 penetrating trauma. As to ISS, it was discovered that 53.0% of the blunt trauma patients had mild trauma (ISS 1-15), 29.4% moderate trauma (ISS 16-24), and 17.6% showed severe trauma (ISS 25); 34.4% of the penetrating trauma patients had mild trauma, 18.7% moderate trauma and 46.9% severe trauma. The mean and standard deviations relating to the ISS of blut and penetrating trauma patients were, respectively, 14.9 +/- 8.1 and 20.8 +/- 11.0; the respective mortality rates were of 11.8% and 12.5%. PMID- 8731273 TI - [Reliability of the certification of the basic causes of neonatal deaths: implications for the study of preventable mortality]. AB - The main causes of neonatal mortality, the reliability of the underlying cause of death registered in the death certificate, and the impact of problems of reliability on the analysis of preventable death were studied. The information on death certificates from a 15% sample of neonatal deaths between May 1986 and April 1987 in the Greater Metropolitan Region of Rio de Janeiro was compared to the information in the hospital records of the 452 deceased infants. A "modified underlying cause" considered most correct according to disease classification rules was identified from the records. The great majority of deaths (87%) were due to perinatal causes. Agreement between the originally declared and modified underlying causes of death was poor: 38% for 3 digits of the International Classification of Diseases Codes (CID-9) and 33% for 4 digits. The modified underlying causes are more weighted towards maternal conditions and complications, which increased by a factor of 12.8, and towards complications of the placenta, umbilical cord, labour and delivery, which rose by a factor of 6.2 in relation to the original causes. The utilization of the "modified" underlying cause elevated considerably (58%) the proportion of deaths considered reducible by the classification of neonatal death proposed by the SEADE Foundation. Seventy five percent (75%) of deaths were considered reducible or partially reducible. One hundred and seven (24%) of the deaths of them being in infants of normal birthweight, of which 60% considered preventable. Four (4) deaths from congenital syphilis, 3 from perinatal hemolytic diseases, and 21 unattended home deaths of infants were also identified. In summary, important problems were identified in the reliability of the declaration of the underlying causes of neonatal death, whose correction tends to elevate the proportion considered reducible or preventable. The potential for the use of death certificate data for the monitoring of quality is evident, nonetheless improvements are needed in the quality of these data. PMID- 8731274 TI - Cost of diseases in Brazil: breast cancer, enteritis, cardiac valve disease and bronchopneumonia. AB - The results from the need to develop methodologies for performing cost analysis in developing countries, principally in the region of Latin America, were studied. It, furthermore, serves to generate knowledge from an economic evaluation in order to support decision-making related to the organization of health systems, particularly in the efficient use of resources which are allocated for the provision of medical services. Two chronic diseases (breast cancer and cardiac valve disease) and two infections (enteritis and bronchopneumonia) were selected for the study. The results recommend the use of a valid methodology for economic cost analysis of any disease to be studied and the use of this information in the decision-making process. PMID- 8731275 TI - [Mood disorders among inpatients in ambulatory and validation of the anxiety and depression scale HAD]. AB - The revised Clinical Interview Schedule (CIS-R) and the Hospital Anxiety and Depression (HAD) Scale were used to estimate the prevalence of mood disorders among 78 consecutive admissions to a general medical ward in a university general hospital in Brazil (43 males and 35 females; mean age = 43.2yr). Interviewers also completed a 5-point symptom severity scales for anxiety and depression. The definition of cases of anxiety [and depression] was based on two criteria: a. score > or = 2 on the CIS-R section of anxiety [> or = 4 on the CIS-R sections of depression and depressive ideas]; and b. score > or = 2 on the clinical severity scale for anxiety [score > or = 2 on the clinical severity scale for depression]. A 39% prevalence rate of affective disorders was found. Sixteen (20.5%) patients met criteria for anxiety, most of the disorders being of mild severity. Twenty sic patients (33%) were depressed, 7 of them in a moderate degree. The HAD was easily understood by the patients. Anxiety and depression subscales had internal consistency of 0.68 and 0.77, respectively. At a cut-off point of 8/9 sensibility and specificity were 93.7% and 72.6% for anxiety, and 84.6% and 90.3% for depression. HAD items correlated positively with the respective subscales. To a lesser degree, they also correlated with the alternative subscale. Our findings confirm the high prevalence of mood disorders among medical in-patients. In clinical practice, the HAD may have a useful role in detecting those patients requiring further psychological care. PMID- 8731276 TI - [Dental health status of students living in places supplied with drinking water of very high and very low levels of fluorides]. AB - The results of a cross sectional epidemiological survey for the purpose of evaluating the state of dental health of schoolchildren (aged 6-7 and 12-13) living in Sampacho and Portena, two towns in the Province of Cordoba (Argentina), supplied with drinking water containing quite different levels of fluoride, are described and analized. In Sampacho, F- level is 9.05 mg/l. while in Portena the concentration is of 0.19 mg/l. The proportion of schoolchildren (aged 6-7 and 12 13) without caries was significaticantly higher in Sampacho than in Portena, while the dmf-t, dmf-s, DMF-T and DMF-S indexes were considerably higher in the latter place. The severity of caries in children (age 12-13) living in Sampacho ranged from low to moderate (DMF-T = 2.53), whilst in Portena the range went from moderate to high (DMF-T = 4.41). No cases of dental fluorosis were recorded in Portena; but in Sampacho, there was a high proportion of children with mild fluorosis (aged 6-7) and mild or severe fluorosis (aged 12-13). Calcium, phosphate, thiocyanate, protein and secretory Ig A salivary levels were similar not only among schoolchildren of both towns but also among those with different experience of caries and different degrees of severity of fluorosis. It is concluded that urgent (preventive and/or curative) sanitary measures are necessary for the purpose of reducing or controlling caries in Portena as well as dental fluorosis in Sampacho. PMID- 8731277 TI - [Hair mercury levels in different occupational groups in a gold mining zone in the north of Colombia]. AB - Hair mercury analysis was carried out on a sample of 219 people living in the main gold mining zone of Colombia, 27 inhabitante of Cartagena City being taken as control sample. For data analysis the sample was divided by occupation and the corresponding the hair mercury concentrations (mean +/- SD) were found to be 5.23 +/- 5.78, 2.83 +/- 3.27, 2.4 +/- 2.02 and 1.33 +/- 0.74 micrograms/g for fishermen, miners, people of various other activities and the control sample, respectively. According to variance analysis and the Newman Keuls test, there were significant differences (p < 0.01) between the mercury concentrations for fishermen and those for the other groups. No significant differences were found for hair mercury and sex, non was any correlation with age detected; however, a low positive correlation (R = 0.15, p < 0.01) with the frequency of the consumption of fish was noted. The main symptoms of mercury poinsoning observed in the persons exposed were headache, oral lesions, metalic taste, loss of memory, and irritability. PMID- 8731278 TI - [Epidemiology of accidents due to bites of poisonous snakes: a study of cases attended in 1988]. AB - The attendance given to patients by ta specialized Hospital, in S. Paulo, Brazil, during 1988 is studied. The study is based on the medical records of 322 patients and on questionnaires filled out by author during interviews with 209 patients or their companions. The 322 snake-bites occurred mainly between October and April, in the diurnal period, mainly in the afternoon. Most of patients were adult males, mainly between 10 and 20 years of age. The parts of the body most frequently affected were the feet, hands and legs. The snakes of the genera Bothrops, Crotalus and Micrurus were responsible, respectively, for 306 (95.0%) 14 (4.4%) and 2 (0.6%) of the accidents under study. Among the 160 snakes that were classified at the Herpetological Section of the IB, 152 were Bothrops; 142 B. Jararaca, mostly young reptiles, and 8 were of the genus Crotalus. Of the patients, 90.4% recovered completely, 2.2% presented sequelae, 7.5% were transferred and thus it was impossible to follow them up. Of the 209 persons interviewed, the occupational group most prone to snake bites was agricultural workers, followed by studentes; nearly 60% of the accidents ocurred during work; most of the patients had their inferior extremities unprotected at the moment of the bite. On hundred and sixty patients (76.6%) submitted to some from of treatment before coming to the HVB-IB, the more common being the use of a tourniquet (50.2%), local squeezing in an attempt to remove part of the venom (33.5), application of substances on the site of the snake bite (36.8%) and the ingestion of others (12.9%). Slightly over a quarter of the patients underwent some kind of medical treatment before coming to the HVB-IB, the most common being antissepsis (8.2%), administration of antivenom (6.2%), antihistamines (5.7%) and analgesics (5.3%). The snake was seen before it struch by 187 (89.5%) of the 209 persons interviewed and in most cases it adopted the strike posture just before the first bite. PMID- 8731279 TI - [Isolation of thermotolerant species of Campylobacter from 2 populations of chickens bred in confinement and at liberty]. AB - The isolation rates of thermotolerant Campylobacter species in free-ranging domestic chickens and confined chickens from Iquitos city, Peru, were determined. Campylobacter spp. were isolated in 54.0% of the former group of chickens, being less frequent (35.0%) in the latter (p < 0.05). Of the classical thermotolerant species, C. jejuni and C. coli were the most frequent. However, the presence C. lari suggests that the chickens might be an important reservoir of this bacterium. PMID- 8731280 TI - [Rabies in insectivorous (Mollossidae) bats of southeastern Brazil]. AB - Four rabid bats belonging to three species of Molossidae (one Molossus molossus, one Nyctinomops laticaudatus and two N. macrotis) were captured in the State of S. Paulo, southeastern Brazil. Three of these bats were found during the day in unusual, visible places and the other came flying through a window in the evening. A descriptive table containing data on 19 similar cases, belonging to eight species, is presented and suggests that bats with atypical behavior must dearly be suspected of being sick, possibly as having rabies. PMID- 8731281 TI - [A modern view of the evolution of virulence]. AB - According to the prevailing, traditional view parasites should develop reduced virulence towards their hosts, because more virulent pathogens are more likely to drive the hosts, and thus themselves to extinction. Virulence is considered to be a primitive stage of a parasitive-host association. However the usefulness and validity of this view have been questioned. Recent studies suggest that parasites need not necessarily evolve towards reduced virulence. The points of view of Darwinian medicine in the direction of the evolution of virulence there may be many possible coevolutionary trajectories, depending on the details of the parasite's life-history, the host's behavior and the transmissibility of the parasite. Theoretical and epidemiological evidences indicate that pathogens transmitted by arthropod vectors are significantly more lethal to humans than those transmitted by personal contact. Water borne enteric pathogens are less virulent after purification of water supplies. Recent experiments also support the emerging theory that parasitism can evolve to be either more or less virulent in a long-term host, depending on the way the parasite is transmitted to the host and on the environment in which they live. PMID- 8731282 TI - [The Brazilian economy in the 80's and its impact on the living conditions of the population]. AB - In the 80's the Brazilian economy underwent one of the most severe crises of its history, resulting in the stagnation of the gross national product and inflation rates such as never previously reported. Despite this unfavorable scenario social indicators have presented a positive evolution. This work shows that although the Brazilian family has adopted the over-use of the family work force as a strategy for facing this crisis on the work market, the evolution of both income and poverty in that period were poor. The increase of expenses and the transformation of the forms of implementation of public policy in the health and nutrition areas are shown to be decisive factors in the performance of social indicators. PMID- 8731283 TI - [Models of causal inference: critical analysis of the use of statistics in epidemiology]. AB - The foundations on which the concept of risk has been constructed are discussed. A description of Rubin's model of causal inference, which was first developed in the domain of applied statistics, and later incorporated into a branch of epidemiology, is taken as the starting point. Analysis of the premisses of causal inference brings to light the logical stages in the construction of the concept of risk, allowing it to be understood "from the inside". The abovementioned branch of statistics and epidemiology seeks to demonstrate that statistics can infer causality instead of simply revealing statistical associations; the model gives the basis for estimating that which way be defined as the effect of a cause. Using this procedural distinction between causal inference and association, the model also seeks to differentiate between the epidemiologial dimension of concepts and the merely statistical dimension. This leads to greater complexity when handing the concepts of interation and coofounding. The redective aspects inherent in this methodological construction of risk are here high lighted. Thus, whether applied to individual or populational inferences, this methodological construction imposes limits that need to be taken into account in its theoretical and practical application to epidemiology. PMID- 8731284 TI - Molecular medicine and clinical epidemiology: new era, new choices. PMID- 8731285 TI - Papillomavirus in cervicovaginal smears of women infected with human immunodeficiency virus. AB - It has been described that women infected with the human immunodeficiency virus (HIV) present more frequent cytological abnormalities in cervicovaginal smears, generally related to infection by human papillomavirus (HPV). The present work is a study of cervicovaginal smears of 147 HIV-seropositive women submitted to routine gynecological examinations. The smears were stained by the Papanicolaou method. Cytopathic effects of HPV were found in 38 (25.8%) cases. Nuclear atypias of cervical intraepithelial neoplasia (CIN) were evident in 36 (24.5%) of these cases: 27 (18.4%), CIN I; 6 (4.0%), CIN II and 3 (2.0%) CIN III. Also 2 (1.4%) invasive carcinomas and one (0.7%) endocervical dysplasia were found. Other agents observed were: Candida sp, 19 (12.9%) cases, Gardnerella vaginalis, 19 (12.9%), Trichomonas vaginalis, 13 (8.4%), Chlamydia trachomatis 5 (3.4%), Mobiluncus sp 2 (1.4%) and Herpes simplex virus 1 (0.7%). This study emphasizes the high frequency of HPV/CIN cervicovaginal abnormalities in HIV-seropositive in our population. It is possible that immunological factors and sexual promiscuity are involved in this phenomenon. PMID- 8731286 TI - Effects of simple hysterectomy on bone loss. AB - Lumbar spine and proximal femoral bone densities of Caucasian women, aged 35-45, were measured by dual photon densitometry model DPX. The measurement sites were assessed at the lumbar spine (vertebrae L2 to L4) and at the proximal femur (trochanter, femoral neck and Ward's triangle). After exclusion of women with climacteric symptoms, sterilized patients or those with menopausal concentrations of gonadotrophins, the study included 22 subjects: 11 menstruant (control group) and 11 hysterectomized. The hysterectomies were without oophorectomy and had been performed during the previous five years. The bone densities of the hysterectomized women were lower than those of the normal ones, but significantly lower at the Ward's triangle. PMID- 8731287 TI - Ostomy or intestinal anastomosis in cases of peritonitis. AB - Twenty-six patients showing peritonitis due to nontraumatic acute abdomen were submitted to ostomy. Mean age was 51 years (range 25-83), being 13 males and 13 females. Bowel obstruction (BO) was the most frequent cause of peritonitis (11 cases), followed by intestinal perforation (IP) (8 cases), acute mesenteric infarction (AMI) (5 cases), and acute abdomen of inflammatory/infectious origin (AAIO) (2 cases). Brook's ileostomy was performed on 65% of the patients. Jejunostomy was performed only in 4 patients, leading to a bad evolution. Overall mortality was 54%. Primary ostomy or anastomosis in cases of peritonitis constitute a highly controversial theme. Indications and problems involving the intestinal exteriorization in emergency surgery urgency are herein discussed. PMID- 8731288 TI - Transsphinctericanorectal reconstruction of ambiguous genitalia: an innovative approach to neovaginoplasty, pioneer in Brazil. AB - The authors, based upon previous experience in the treatment of ten children with cloacal abnormalities, among which two with female pseudohermaphroditism with cloaca, hereby present a pioneer experience in Brazil, with the "posterior sagittal transsphinctericanorectalurethrovaginoplasty" a surgical approach to neovaginoplasty described by "Cripps/Pena", which was utilized for the reconstruction of the genitalia of three intersexuated adolescents with protective colostomy, who presented an urogenital sinus, high vaginal implantation and a normal rectum. One patient was an adrenal female pseudohermaphrodite and two were male pseudohermaphrodites. The authors present a brief report on the three cases, describe the surgical procedure, relate on the anatomy of the urogenital sinus and discuss surgical indications, diagnosis and results, comparing them to the reviewed international literature. PMID- 8731289 TI - Large placental hemangioma diagnosed by ultrasonography--a case report. AB - We present a case of large placental hemangioma comprising more than half of the organ and not causing feto-maternal complications. It appeared after 29 weeks of gestation. At 29 weeks an ultrasonography disclosed a normal placenta. At 35 weeks of gestation it measured 60 x 57 mm and appeared as a well delineated hypoechoic image. Delivery took place at 38 weeks by cesarean section and the child was normal. PMID- 8731290 TI - Current concepts on the transmission of bacteria and parasites by blood components. AB - Several bacterial and parasite transfusion-transmitted diseases have been described in the medical literature. This review deals with the main bacterial (Syphilis, Lyme disease, Gram positive and Gram negative agents), parasite (Chagas disease, malaria, leishmaniasis, toxoplasmosis and babesiosis) and rickettsial diseases that are carried by blood products. Preventional aspects (e.g. storage, screening tests, use of leukocyte-depleted components), diagnosis, geographical distribution and the incidence of these transfusional hazards are also discussed. PMID- 8731291 TI - Hypertonic volume therapy: feasibility in the prevention and treatment of multiple organ failure and sepsis. AB - Small-volume resuscitation by means of bolus infusion of hypertonic saline solutions was first applied for the primary treatment of severe hemorrhagic and traumatic shock and promptly restored central hemodynamics and regional organ blood flow. Mechanisms of action are diverse--i. maintenance of high cardiac output (direct myocardial stimulation; increase in intravascular volume); ii. maintenance of peripheral arterial vasodilation (effect of hyperosmolality; plasma volume effect) and iii. reduction of tissue edema (shifting of tissue water along the osmotic gradient). These mechanisms promote the restoration of the severely impaired microcirculation frequently seen also in sepsis. Hypertonic volume therapy has been the object of several experimental studies of acute hyperdynamic endotoxemia, however, a greater number of clinical studies have to be developed for the better understanding of the positive, and perhaps hazardous, effects of small-volume resuscitation in sepsis and multiple organ failure. The aim of this paper is to review the concepts involving such solutions, and their potential use in treatment of profound hypovolemia and microcirculatory deterioration associated with sepsis and endotoxic shock. PMID- 8731292 TI - A mutation affecting gluconate catabolism in Escherichia coli: the locus for the main high affinity transport. AB - The bioH-malA region of the E. coli chromosome (min 75.5) includes the gntT gene which encodes a high affinity transport for gluconate. Other gnt loci have not been characterized in this region; nevertheless, because lesions in it affect severely the utilization of gluconate, it has been suggested as being more complex. This region was investigated with respect to gluconate catabolism through the characterization of suitable E. coli strains lysogenized with a specialized transducing phage carrying the bioH-malA region of the bacterial chromosome (lambda cI857st68h80d2bioH-malA). It was found that the region transduced by this phage while includes the gntT gene lacks other gnt loci that might code additional activities for transport of gluconate or its phosphorylation. Moreover, the pleiotropic lesion gntM2, previously mapped into this region and suggested as altering gntT or a presumptive regulator gene that might be involved in this catabolism, resulted recessive in lysogens (partial diploids) containing the defective prophage. The results obtained supported the idea that gntM2 is an allele of gntT; consequently those results suggested the precise position of this gene on the cromosomic map and the central role that its product might have in the initial incorporation of gluconate in E. coli. PMID- 8731293 TI - [Effect of the administration of furosemide on the fluid filtration rate and in pulmonary artery pressure in isolated lungs of rabbits]. AB - To study the possible effects of Furosemide at the lung level, two groups of isolated rabbit lung preparation were studied. An experimental group underwent a pulmonary hydrostatic oedema when the pressure of the left auricle (PAI) was increased from 0.45 +/- 0.74 t0 11.8 +/- 2.9 cm of H2O, with that increase in PAI we obtained an increase of 0.457 +/- 0.51 g/min in FFR (Fluid Filtration Rate), during this stable and sustained oedema, a 2 mg/Kg dosis of Furosemide was injected every 10 minutes and the possible changes in PAP, PAI, PVA, TFL, PaO2, PaCO2 and pH was observed, but no changes were observed in these parameters during the Furosemide infusion, and the same effect was observed in the control group were the preparations were maintained in basal conditions and without oedema. These results suggests that the Furosemide hat not a direct cardio pulmonary effects, and the only possible effects could be by increasing diuresis at renal level. PMID- 8731294 TI - Detection of microcin production by pathogenic Escherichia coli isolates from children with acute diarrhoea. AB - Production of low molecular weight antibiotic substances was detected among pathogenic Escherichia coli isolates from diarrhoeal children's feces during evaluation of a rotavirus vaccine in Caracas city. One of these products, microcin V627a, was partially purified and characterized. Microcin synthesis and immunity system appears not to be plasmid determined and the antibiotic was produced to a high level in minimal medium during the stationary phase. This microcin has many features in common with the mccA15 family. PMID- 8731295 TI - [Effect of the in vivo administration of beta-methyldigoxin on the Na, K-ATPase measured in different tissues of guinea pigs]. AB - We measured the activity of the Na,K ATPase, Mg dependent and inhibited by ouabain, in microsomal fractions obtained from guinea pig myocardium and kidney, as weel as red cell ghosts. A group of animal was treated with beta-methyl digoxin (0.2 mg intraperitoneally) about one hour before obtaining the tissues. The control group received no medication. The plasma of both group, control and treated, had similar potassium concentrations (4.3 +/- 0.87 mM and 4.3 +/- 0.66 mM, respectively). The plasma digoxin concentrations from treated animals was always high (76.4 +/- 34.1 ng/ml). The Na,K-ATPase activity in the microsomal fraction from treated animals decreased by 28.7% in myocardium and by 27.7% in red cell ghosts, in comparison to the enzime activity measured in control animals. On the other hand, the Na/K activity obtained in kidney microsomal fraction did not change with treatment. We then measured the Na,K-ATPase activity in the red cell ghosts and microsomal fractions obtained from myocardium and kidney of untreated Guinea pigs. Adding digoxin in vitro (1 x 10(-9) M to 1 x 10( 3) M) we obtained, in myocardial fractions, a 50% maximal inhibition; the digoxin concentration causing half maximal effect (DI50) was 7 x 10(-5) M. In the kidney microsomal fraction we measured a 66% maximal inhibition of the enzime activity and DI50 was 2 x 10(-6) M. For red cell ghosts the maximal enzime inhibition was 34% and the DI50 was 1 x 10(-5) M. In conclusion, in the Guinea pig, the acute in vivo administration of beta-methyl digoxin causes a parallel inhibition of the Na,K-ATPase from myocardial fractions and red cell ghosts. We measured no significant change in the kidney microsomal fractions. We propose the determination of red cell Na,K-ATPase activity as possible indicator of digitalis effect on humans treated with these drugs. PMID- 8731296 TI - [Skeletal muscle ultrastructural pathology in bovine paraplegic syndrome]. AB - This study reports the ultrastructural alterations found in biopsies taken from fast twitch gastrocnemius and slow twitch soleus muscles from bovines affected by the paraplegic syndrome. The abnormalities observed were contractile and sarcotubular systems disorganization, proliferation of autophagic vacuoles with mitochondrial material and glycogenosomes, decrease of nuclear chromatin electron density, and sacrolemmal damage in areas of segmental necrosis where macrophages were also seen. Capillary alterations were slight. Mentioned abnormalities are discussed in connection with mineral nutritional deficiences presented by the studied animals. PMID- 8731297 TI - [Determination of micro and macronutrients in the cattle of the Venezuelan plains and their influence on the origin of bovine paraplegic syndrome]. AB - We report a study carried out in three livestock-producing regions of Venezuela to determine the mineral status of grazing cattle and its relationship to the Sindrome Parplejico del Bovino (SPB). Animal tissue samples from blood and liver were collected from a total of 17 farms within three regions: southwest (Apure), central (Guarico) and southeast (Bolivar) both during the dry and rainy seasons. In SPB free animals, the serum levels of sodium, potassium, chloride, magnesium, total and ionized calcium, phosphorus, and creatinine, were within the normal range. Glucose was found to be deficient in cattle from Bolivar and Guarico states and normal in Apure. With the exception of liver copper and serum zinc, all the other microelements analyzed (liver cobalt, and molybdenum, and serum iron) were found to be normal. Copper was found to be low in all regions studied with a mean value of 74.8 ppm indicating a moderate deficiency of this element. Similarly, in the central and southwest regions, zinc was found to be close to 0.34 ppm, significantly lower than the critical level of 0.7 ppm. In order to determine the effect of the dry and rainy seasons on the content of macro and microelements, controlled group of cattle from the three regions were followed in their contents of magnesium, calcium, copper and iron. In the dry season all of these elements tended to be much lower, showing a significant increase in the rainy season. This increase was much greater in cattle that received mineral supplementation and sanitary treatment. Bovines with diagnosis of SPB showed: low liver copper content, low serum magnesium and phosphorus levels significantly higher that control cattle. PMID- 8731298 TI - [The role of low-molecular-weight heparin in unstable angina, acute myocardial infarction and post-elective percutaneous transluminal coronary angioplasty]. AB - PURPOSE: To evaluate the clinical efficacy and safety of subcutaneous (SC) low molecular weight heparin (LMWH) compared to intravenous (IV) non fractioned heparin (NFH) in unstable angina, acute myocardial infarction and post percutaneous transluminal coronary angioplasty. METHODS: From September/92 to April/94, 314 patients were randomized in two groups. Group I-- 154 patients treated with SC LMWH, using in the 1st phase SC LMWH with a dosage of 160 UaXa IC/kg/day (group IA--92 patients), and in the 2nd, a dosage of 320 UaXa IC/kg/day (group IB--62 patients). Group II--160 patients treated with IV NFH 100UI/kg (bolus), followed by 1000UI/h with adjusted dosage by activated partial thromboplastin time. RESULTS: There was not a statistically significant difference among the three groups in relation to cardiac events, hemorrhagic complications and deaths. CONCLUSION: The clinical efficacy and safety of SC LMWH in patients with unstable angina, acute myocardial infarction and post percutaneous transluminal coronary angioplasty were similar to IV NFH with the dosages used in this study. PMID- 8731299 TI - [Assessment of the myocardial functional reserve in patients with left ventricular aneurysm by radionuclide ventriculography. Response to isotonic exercise]. AB - PURPOSE: To evaluate global and regional left ventricular (LV) ejection fractions (EF) by radionuclide ventriculography in patients with LV aneurysm at rest and during isotonic exercise. METHODS: Twenty patients were studied by radionuclide ventriculography at rest and during exercise. All patients had been submitted to cineangiography and showed LV aneurysm post myocardial infarction. RESULTS: Patients were divided according to LV EF in two groups: one with EF > or = 40% and the other with < 40% EF. Both groups showed normal response of global EF to exercise: mean rest EF was 40 +/- 14% and mean exercise EF was 45 +/- 14% (p < 0.01). When groups were considered separately, EF values showed the same behavior. Half of the patients showed normal response to exercise and the other half showed abnormal response. These changes were not associated with resting EF values, but were due to regional EF of lateral wall, that changed from 44 +/- 7 to 48 +/- 7% in the group of patients with normal LV EF response to stress and from 50 +/- 5 to 46 +/- 5% in those with abnormal response (p < 0.01). CONCLUSION: The evaluation of regional ventricular EF by radionuclide ventriculography during exercise better discriminates functional reserve in patients with LV aneurysm than resting global EF. These findings could help the decision making of the therapeutic approach in this specific group of patients. PMID- 8731300 TI - [Risk factors in university students]. AB - PURPOSE: To evaluate the presence of risk factors (RF) for coronary disease in a group of university students (US). METHODS: We studied 123 US, 88 (71.55%) females, mean age 21 +/- 3.52 years, submitted to questionary, interview, physical examination and serum analises. Eighty six women informed about oral contraceptive use. Serum lipid levels were measured in 66 women and 21 men. Statistical analysis was proceded by analysis descritive. RESULTS: Sedentarism was observed in 51.14% and smoking in 19.32%; this two RF were observed in 5.71% of males. In 86 women, 53.49% never used oral contraceptives, 17.44% used it in the past and 29.07% were presently in use. Family history of cardiovascular diseases, was unknown in 34.95% of US. Systolic arterial pressure was higher in men. In 22.99%, cholesterol (C) and LDL-C were, respectively higher than 200mg/dl and 130mg/dl; HDL-C were low of 35mg/dl in 2.3%. There were no differences between the two sexes in the dosages of HDL-C. CONCLUSION: RF were frequent in the studied group and smoking, sedentarism and dislipidemy are larger prevalent. Preventive measures are important because can be controlled or removed. PMID- 8731301 TI - [Anatomic variations of the coronary arteries]. AB - PURPOSE: To study the coronary arteries and their main branches showing the aspects of source, trajectory and anastomoses of these vessels at the subepicardial level. METHODS: The study was carried out on 110 adult human hearts, of both sexes, fixed in 10% formaldehyde solution. The pericardium was removed to expose the coronary arteries and their branches at the subepicardial level. RESULTS: In 38.18% of the cases the left coronary artery presented a trifurcation into anterior interventricular, circunflex and left marginal branches (35.70%) and into anterior interventricular, circunflex and lateral branches (64.30%). In 60% of the hearts examined, the left coronary artery presented a bifurcation into anterior interventricular and circunflex branches. In 1.82% of the cases these two branches arise directly from the aorta. An anastomosis, at the subepicardial level, between the anterior and posterior interventricular branches was observed in 56.36% of the hearts. In 88.18% the posterior interventricular branch arised from the right coronary artery, whereas in 11.82% this vessel arises from the circunflex branch. Anastomoses between the right coronary artery and the circunflex branch were found in 10% of the hearts (crux cordis). The dominance of the right coronary artery was present in 69.09% of the cases, of the left coronary artery in 11.82% and in 19.09% of the hearts had balanced distribution. CONCLUSION: The coronary arteries and their main branches present a great quantity of variations with regard to source, trajectory and anastomoses. This knowledge is important for the interpretation of coronary angiography and surgical myocardial revascularization. PMID- 8731302 TI - [Prevalence and sociocultural and economic aspects of hypertension in a health center of the northern area of Manaus]. AB - PURPOSE: To verify the prevalence and the possible sociocultural and economic factors involved in high blood pressure (HBP), intending to contribute to the elaboration of public health programs. METHODS: The blood pressure of 1,766 individuals, aging 13 years old or more, was measured by the auscultatory method, using a stethoscope and a sfigmomanometer. It was included in the HBP group, the individual presenting blood pressure above 140 x 90 mmHg in two measurements on different occasions. These individuals answered a form searching for sociocultural and economic aspects. RESULTS: Among the individuals examined, 76 were considered to have HBP. There was an increase on the prevalence of HBP with an increase of age, being the greatest among the individuals aging 49 years or more. It was observed significant association between age and HBP (p < 0.05), but the same did not happen with sex or skin color. In most part of the individuals with HBP, it was observed low family income and low educational level. Half of the HBP patients were used to eat with salt and fat. CONCLUSION: The educational level, economic status, and sociocultural habits of a person constitute factors that must be considered in the development of public health programs for control and reduction of the prevalence of HBP. PMID- 8731303 TI - [Tuberculous pericarditis as the initial manifestation of acquired immunodeficiency syndrome]. AB - The most common externalization of the acquired immunodeficiency syndrome (AIDS) is opportunist infection, and tuberculosis is one of the most frequent agents. The tuberculous pericarditis has been associated with AIDS, but it exceptionally occurs as the first event in the syndrome. We reported four cases in which tuberculous pericarditis was the initial manifestation of AIDS, characterizing by its clinical picture, diagnostic methods, therapeutics and the evolution of this involvement. PMID- 8731304 TI - [Case 6/95 (Instituto do Corac-ao do Hospital das Cl-inicas-FMUSP)]. PMID- 8731305 TI - [Ventricular assistance by direct pneumatic massage]. PMID- 8731306 TI - [Dilated cardiomyopathy. Is its etiology still unknown?]. PMID- 8731307 TI - [The role of low-molecular-weight heparin in unstable angina, acute myocardial infarction and post-elective percutaneous transluminal coronary angioplasty]. PMID- 8731308 TI - [Morphologic aspects of the left ventricular remodeling in hypertensive cardiomyopathy]. PMID- 8731309 TI - [Echocardiographic criteria in the diagnosis and functional evaluation of hypertensive heart disease]. PMID- 8731311 TI - [Hypertrophic response to cardiac overload. Similarities and differences between left ventricular hypertrophy as response to pressure overload or to volumetric overload]. PMID- 8731310 TI - [Heart hypertrophy. Determinant factors and molecular mechanisms]. PMID- 8731312 TI - [Arrhythmogenic potential of hypertensive cardiomyopathy. Prognostic value of Holter monitoring]. PMID- 8731313 TI - [Left ventricular hypertrophy and cardiovascular risk. Critical analysis of the use of diuretics and adrenergic beta blockers]. PMID- 8731314 TI - [Pharmacologic regression of cardiac and vascular structural changes in the hypertensive patient. A question that goes beyond the exclusive medical bias]. PMID- 8731315 TI - [Infective endocarditis in 100 patients subjected to heart transplantation]. AB - PURPOSE: An analysis of occurence, etiology, clinical aspects and death rate of infectious endocarditis cases involving patients who underwent heart transplantation. METHODS: 100 consecutive heart transplant patients were analysed; follow-up varied from three to 90 (medium of 25.38, SD +/- 27.97) months. Diagnostic criteria for endocarditis were those of the epidemiology and quality control service, Heart Institute, Hospital das Clinicas da Faculdade de Medicina da Universidade de Sao Paulo, that agree with those of the Center for Disease Control. Diagnosis was established mostly by blood cultures, echocardiograms, either transthoracic or esophageal and autopsy. RESULTS: Six cases of endocarditis were recognized, with four deaths; in three of those we found severe systemic compromise. All cases had fever as an important symptom. CONCLUSION: Death risk seems high of infectious endocarditis after heart transplantation. The cause of this high risk appears to be linked to the bacteria themselves, to association with other clinical situations and to the use of immunosuppresive agents. We recommend a high index of suspition when risk factors are present in order to make a rapid diagnosis soon enough in the natural history of the disease; treatment has to be started as soon as possible. PMID- 8731316 TI - [Analysis of elastic retraction in the 1st 15 minutes after coronary balloon angioplasty]. AB - PURPOSE: To determine the time course of elastic recoil (ER) in the first 15min after successful percutaneous transluminal coronary angioplasty (PTCA). METHODS: One hundred and fifty four patients, with stable or unstable angina were successfully submitted to PTCA. Coronary angiography was undertaken shortly after balloon deflation and repeated 5, 10 and 15 min thereafter. Quantitative coronary angiography was performed with the aid of an eletronic caliper. We calculated the minimal luminal diameter (MLD) and elastic recoil in all angiograms. RESULTS: The average artery's reference diameter was 3.09 +/- 0.61mm and the maximal balloon diameter was 2.95 +/- 0.52mm. MLD before the procedure was 0.65 +/- 0.42mm reaching 2.23 +/- 0.55mm immediately after dilatation (p < 0.0001), and decreasing to 2.09 +/- 0.47mm at 5min (p < 0.0001), 2.01 +/- 0.47 at 10min (p < 0.0001) and to 1.91 +/- 0.56mm at 15min (p < 0.0001). ER increased during the 1st 15min after PTCA, averaging 34.29 +/- 20.40%. In the group of patients whose balloon/artery relationship was < or = 1, the total ER was 0.90 +/- 0.74mm at 15min and 1.20 +/- 0.50mm when the ratio was > 1 (p < 0.0001). We noted that ER in the group of patients with residual stenosis ranging from 30 to 50% at the immediate angiogram after PTCA was greater than in the group whose residual stenosis was less than 30%. CONCLUSION: ER is a dynamic and progressive phenomenon taking place within the 1st 15 min after a successful PTCA. Total ER was 34.29 +/- 20.40% at 15min and was greater when balloon/artery relationship was > 1. Residual stenosis ranging from 30 to 50% in the control immediately after the procedure is a predictive factor of greater ER in the 15min following PTCA. PMID- 8731317 TI - [Evaluation of ischemic preconditioning on collateral circulation, ventricular function and clinical outcome in acute myocardial infarction]. AB - PURPOSE: To evaluate the influence of ischemic preconditioning (IP) in collateral circulation (CC), early ventricular function and in hospital outcomes after myocardial infarction (MI). METHODS: We studied 97 patients with a 1st anterior MI within 6h of pain and isolated total proximal occlusion of the left anterior descending artery, divided in 2 groups: with (GA) or without (GB) angina before MI. Coronariography and ventriculography were performed prior to reperfusion. The left ventricular (LV) ejection fraction was measured by the area length method and anterior wall motion by the centerline method. RESULTS: There was no difference between the two groups in sex, age, CKMB level, treatment, reperfusion rate. Global LV ejection fraction and anterior wall motion were similar, respectively, 39 +/- 9% and -2.55 +/- 1.17 SD/chord for GA and 37 +/- 8% and 2.75 +/- 0.79 and -2.75 +/- 0.79 SD/chord for GB (p = ns). The incidence of visible CC to the infarct area was also similar (present in 6 GA vs 8 GB patients). However, GA patients fared significantly better during hospitalization: No GA patient presented Killip class > or = 2 compared to 8 GB patients (p = 0.007). CONCLUSION: Although collateral coronary circulation grades, global and regional LV function were similar between the two groups, the presence of angina pectoris preceding an acute myocardial infarction was associated with a better in hospital evolution, what could be partially explained by preconditioning phoenomena. PMID- 8731318 TI - [Aortic valve replacement with pulmonary autograft (Ross procedure). Initial experience]. AB - PURPOSE: Report the initial surgical experience with four cases utilizing a pulmonary autograft for aortic valve replacement. METHODS: Four patients, all males, white, age between 23 and 46 years having aortic valve disease were submitted to aortic valve replacement with a pulmonary autograft using the root replacement technique. Right ventricular outflow tracts were reconstructed with antibiotic sterilized pulmonary or aortic homografts. All patients had control bidimensional eco-doppler (ECO) and hemodynamic study to evaluate the function of the implanted auto and homografts. RESULTS: All patients had an excellent post operative recovery, without the necessity of inotropic drugs. All presented in normal sinus rhythm. Post-operative ECO and hemodynamic studies revealed excellent function of the implanted autografts, without gradients in three and with a 15mmHg mean residual gradient in one case. There was no regurgitation in three cases and only trace aortic insufficiency in one. The right sided homografts also showed good function, with no gradient in two cases and mean systolic gradient of 6 and 8mmHg in the other two. CONCLUSION: The pulmonary autograft procedure should be implemented definitely in our country. PMID- 8731319 TI - [Reliability of arterial blood pressure measurement in an arterial hypertension study]. AB - PURPOSE: To evaluate the relaibility of the blood pressure measurements in an arterial hypertension study. METHODS: It was evaluated 502 nurse workers of a public hospital. After careful training and selection of the observers for the quality control at the fieldwork, it was analyzed the correlation between the 1st and 2nd measurements, final digit preference and mean systolic (SBP) and diastolic pressure (DBP). 5% of the sample had their measures compared with those checked by an expert. RESULTS: There was a great confiability intra-observer (96.2% in the SBP and 93.4% in the DBP). The final digit distribution was almost identical. At the 5% sample, there was a strong correlation between them (96.0% in the SBP and 93.9% in the DBP). CONCLUSION: This methodology is feasible in national studies and confers validity to the results. PMID- 8731320 TI - [Ventricular fibrillation in a patient with the permanent form of junctional reciprocating tachycardia]. AB - A case of a 20 year old young man having the permanent form of junctional reciprocating tachycardia complicated by ventricular fibrillation (VF) is reported. A number of antiarrhythmic drugs either as single or combined therapy fail to control tachycardia. Paroxisms of a faster palpitation which never lasted longer than a few minutes were felt over the last six months before the occurrence of VF. At admission, a narrow QRS tachycardia 250 beats/min was recorded and soon degenerated into VF. After electrical shock with 350J permanent form of junctional tachycardia resumed. Electrophysiologic evaluation identified an accessory pathway with long conducting times in postero-septal location, enhanced atrioventricular node conduction and inducible atrial flutter. The patient underwent successful radiofrequency ablation and is doing well after 18 months of follow-up, being off antiarrhythmic drug and tachycardia free. PMID- 8731321 TI - [Arterial hypertension due to mercury intoxication with clinico-laboratorial syndrome simulating pheochromocytoma]. AB - A 17 year-old boy was admitted to the hospital because of severe hypertension (200/130 mmHg), headache, irritability, and sweating. Initial biochemical tests suggested pheochromocytoma, being treated with nifedipine, clonidine and propranolol. However, with report of exposure to mercury vapor, twenty-four-hour urine screening and measurement of blood mercury confirmed intoxication. The patient underwent courses of chelation therapy with dimercaprol (BAL) and penicillamine with remission of symptoms and normalization of blood pressure after 2 months. This case has relevance for current practice reflecting similarity between mercury intoxication and hypertension secondary to pheochromocytoma. PMID- 8731323 TI - [Risk factors for arteriosclerosis in postmenopausal women]. PMID- 8731322 TI - [Lipid changes of fibrinogen and of platelet aggregation induced by etofibrate]. AB - PURPOSE: To evaluate modifications on lipid profile, fibrinogen and platelet aggregation induced by etofibrate. METHODS: Twenty-one adult patients were studied. They all had primary hyperlipidemia and had already been on the AHA step I diet and placebo. Etofibrate (500mg/day) was administered for 60 days in the active phase, when lipid parameters, fibrinogen and platelet aggregation were measured. RESULTS: The % significant reductions were: total cholesterol (-9.50%), LDL-cholesterol (-7.88%), triglycerides (-19.07%), total cholesterol/HDL cholesterol(-11.90%), LDL-cholesterol/HDL-cholesterol (-10.20%), fibrinogen ( 12.79%), platelet aggregation with adrenaline (-24.02%), with ADP 1 mumol ( 30.13%), and ADP 3 mumol (-24.51%). CONCLUSION: The beneficial effects of etofibrate were observed not only on the lipid profile but also on the thrombogenic parameters measured by fibrinogen and platelet aggregation. PMID- 8731324 TI - [Infective endocarditis in patients subjected to heart transplantation]. PMID- 8731325 TI - Molecular analysis of CYP21 and C4 genes in Brazilian families with the classical form of steroid 21-hydroxylase deficiency. AB - The most common enzymatic defect of steroid synthesis is deficiency of the adrenal steroid 21-hydroxylase. Inhibition of the formation of cortisol results in an increased pituitary release of ACTH which in turn drives the adrenal cortex to overproduce androgens. This hormonal setting affects the development of genetic females by misdirecting the differentiation of external genitalia towards the male type. Since the isolation of the gene encoding 21-hydroxylase enzyme in 1984, gene deletions, large gene conversions, and microconversions have been reported to be responsible for the disease. In this paper, we report a study of this genetic defect in 22 families with one or more affected offspring diagnosed as having the classical form of congenital adrenal hyperplasia. The DNA from 30 patients was analyzed with three restriction enzymes. Hybridization with a 21 hydroxylase cDNA probe and the 5' end of a C4 genomic probe disclosed gene deletion in 7.3% (3/41) of the disease-related chromosomes. The rate of large gene conversion was 17.1% (7/41), and no abnormality in the hybridization pattern was observed in 75.6% (31/41) of the disease alleles. Densitometry of the autoradiographs was used to determine the ratio of the copy-number of the 21 hydroxylase gene (CYP21B) to the copy-number of its pseudogene (CYP21A). Differences in phenotype, the low frequency of gene deletion, and the high frequency of gene conversion compared with other studies in different populations indicated that 21-hydroxylase deficiency in the Brazilian population may involve different molecular mutations. PMID- 8731326 TI - Poliovirus type 1 isolated from a vaccine-associated case of paralytic poliomyelitis in Brazil. AB - This study reports a type 1 poliovirus strain isolated in Brazil from a case classified as vaccine-associated paralytic poliomyelitis (VAPP). After serotyping of the viral isolate with hyperimmune equine sera, PCR and molecular hybridization techniques characterized the strain as P1/Sabin-derived. The isolate was partially sequenced to identify mutations at nucleotides 480, 525 and 6203, which are important for reversion of the P1/Sabin strain to neurovirulence. In a recent study, a P1/Sabin-derived strain isolated from the central nervous system of a VAPP case did not mutate at these positions, but maintained 480-G and 525-U (and 6203-C), suggesting that these mutations are not essential for the occurrence of disease (Georgescu et al., (1994), Journal of Virology, 68: 8089 8101). Although the Brazilian strain also maintained 480-G and 525-U (and 6203-C) and was isolated from the stool, the possibility that this isolate invaded the central nervous system after replicating in the gut, causing the paralysis, cannot be ruled out. This is the first report of a type 1 VAPP case in Brazil, although some cases caused by type 2 and type 3 strains have been described. PMID- 8731327 TI - Coelom-associated lymphomyeloid tissue (milky spots): site of lymphoid and myelomonocytic cell generation. AB - Pleural and peritoneal milky spots (MS) are small morphofunctional structures representing subsidiary foci of coelom-associated lymphomyeloid tissue (CALT). In this paper we studied the cellular composition of CALT in normal and Schistosoma mansoni-infected mice. In the healthy mouse, CALT is mainly composed of IgM (+) B cells and presents lower numbers of CD23 and CD45R (B220) B2 lymphocytes. When activated by the infection, it may show pronounced lymphocytosis, plasmocytogenesis (IgM > IgG > IgA > IgG2a > IgG1) and myelomonocytosis. The lymphocytes were mainly of the B1 type (double positive CD5/IgM), with smaller number of T cells (TCR alpha beta (+), TCR gamma delta (+), CD3 (+) and CD5 (+)) and conventional B2 cells (B220 (+), CD23 (+)). The myeloid compartment was composed of immature and mature cells of monocyte/macrophage, eosinophil, neutrophil and megakaryocytic lineages, especially in the omental milky spots. CALT is also a favorable microenvironment for LFA-1 (+) mast cells. Thus, CALT appears to be a mixed lymphoid organ, with secondary and/or primary lymphoid organ functions, being an important site of B1 cell generation, plasma cell maturation and extramedullar hematopoiesis. CALT operates as an interface between blood and lymphatic circulation and coelomic cavities, because locally or externally produced cells have easy and ready access to the pleural and peritoneal cavities. Furthermore, MS cells can escape into blood and lymphatic vessels, providing lymphocytes to other lymphoid organs and to the mucosa associated lymphoid tissue. PMID- 8731328 TI - Specificity and role of anti-Trypanosoma cruzi clearance antibodies. AB - Two strains of Trypanosoma cruzi (Y and CL) were used to study the specificity and role of anti-T. cruzi clearance antibodies. Clearance antibodies were only induced after immunization with living blood-stream trypomastigotes (Btrys) but not with dead parasites. Btrys of either strain were readily cleared from the circulation after passive immunization with anti-Y or anti-CL serum provided that the homologous strain was used. CL or Y Btrys sensitized in vitro with the homologous or heterologous antiserum and transferred to normal mice were cleared from the circulation only when the homologous antiserum was used. Clearance antibodies were removed from serum by absorption with the homologous but not with the heterologous strain. Clearance antibodies were removed from serum by absorption with living Btrys but not with fixed parasites. These results suggest that: a) the parasite epitopes involved in the clearance are peculiar to each strain, b) the clearance antibodies are specific to these epitopes, and c) a proper conformation of the parasite antigens is required for the induction and effector activity of the clearance antibodies. PMID- 8731329 TI - Evaluation of two different oxygen inspiratory fractions on the hemodynamic effects of N omega-nitro-L-arginine methyl ester in anesthetized dogs. AB - The effect of two different oxygen inspiratory fractions (FiO2 = 21% and 100%) on the hemodynamic responses induced by N omega-nitro-L-arginine methyl ester (L NAME) was investigated in anesthetized dogs. L-NAME (0.01-10.0 mg/kg), but not D NAME, induced dose-dependent changes in the hemodynamic parameters of the animals. At the highest dose, L-NAME increased mean arterial blood pressure in both room air (from 86.2 +/- 3.2 to 125.1 +/- 7.8 mmHg) and pure oxygen (from 100.0 +/- 7.5 to 139.0 +/- 3.2 mmHg) ventilated animals. L-NAME also increased systemic and pulmonary vascular resistances. These effects were accompanied by a decrease in cardiac output and bradycardia (37% and 31% decreases for pure oxygen and room air, respectively). However, there were no significant differences in the responses to L-NAME between the dogs ventilated with FiO2 = 21% and those ventilated with FiO2 = 100%. L-NAME did not modify blood gas analyses, despite the expected difference in pO2 levels between the two experimental groups of animals (3 times higher in the animals ventilated with pure oxygen). These results indicate that nitric oxide release accounts for the maintenance of hemodynamic function in the anesthetized dog, and that L-NAME-induced effects are not affected by hyperoxemia. PMID- 8731330 TI - Food deprivation and hypothermia in desynchronized sleep-deprived rats. AB - Reports of the effect of desynchronized sleep (DS) deprivation on body temperature (Tb) of rats in the literature are contradictory. Since conspicuous body weight loss is common in such deprivation, the effect of food plus DS deprivation on Tb of adult male Wistar rats was studied. DS deprivation carried out by the small platform method with food ad libitum (N = 8) induced hyperthermia (Tb above 38.5 degrees C) in 1 to 3 rats daily until the 8th day, when a case of discrete hypothermia (Tb below 36.9 degrees C) appeared. Food deprivation alone started to induce hypothermia on the third day in one (20%) out of five rats. Fasting imposed from the 5th to the 8th day of DS deprivation (N = 12) caused hypothermia in 33% and 67% of the animals on the second and third day of starvation, respectively. DS compensatory manifestations in 6 starved rats intensified (N = 2) or precipitated (N = 2) hypothermia after the end of sleep deprivation. It is concluded that the hypothermia is not a primary effect of DS deprivation, and this state of sleep seems to have its particular functional role which is independent of thermoregulation. PMID- 8731331 TI - Effect of nifedipine on arterial pressure lability of sino-aortic deafferentated rats. AB - Sino-aortic deafferentation (SAD) in rats produces arterial pressure lability (APL) whose mechanisms are not completely understood. In the present study we infused nifedipine (500 micrograms kg-1 min-1) for 30 min into sham-operated and acute SAD rats (24 h) and the changes in mean arterial pressure (MAP) and APL were measured during this period by a computerized technique. APL was reported as the standard deviation of the mean of all MAP sampling points. In SAD rats with high basal APL levels (SAD high APL, N = 8) nifedipine produced a significant reduction in MAP (from 116 +/- 5 to 88 +/- 3 mmHg) and APL (from 19.62 +/- 1.46 to 9.51 +/- 0.88 mmHg), while in SAD rats with low basal APL levels (SAD low APL, N = 8) it significantly reduced MAP (from 120 +/- 7 to 95 +/- 6 mmHg) and did not change APL (9.46 +/- 0.69 to 10.93 +/- 1.62). Nifedipine infusion into sham operated rats (N = 8) produced no significant changes in MAP (105 +/- 3 vs 104 +/ 2 mmHg) or APL (4.17 +/- 0.53 vs 3.99 +/- 0.67 mmHg). Since nifedipine infusion reduced APL only in SAD high APL rats but reduced the MAP in both groups of SAD rats, we conclude that its effect on APL is not related to decreases in MAP. These data suggest that extracellular calcium influx through calcium channels plays an important role in the generation of APL, especially in SAD rats with high APL levels. PMID- 8731332 TI - Lower transversal laparotomy has no appreciable effect on respiratory mechanics in guinea pigs. AB - Respiratory system, lung, and chest wall resistances and dynamic elastances were determined in six anesthetized, paralyzed, and mechanically ventilated guinea pigs both before and after lower transversal abdominal opening performed at the level of the spina iliaca anterosuperior. Furthermore, the resistances were also split into their two components, one reflecting the Newtonian resistances and the other representing the viscoelastic/inhomogeneous pressure dissipations in the system. The method of end-inflation occlusion during constant inspiratory flow was used. Chest wall configuration was also evaluated by measurements of lateral and anteroposterior diameters and circumferences at the 4th intercostal space and xiphoid levels both at functional residual capacity and at the end of tidal inspiration before and after surgery. After abdominal incision no statistically significant changes could be detected in any of the measured variables. It may be concluded that lower transversal abdominal opening does not alter respiratory mechanics. PMID- 8731333 TI - General introduction to chronobiology. AB - Rhythmicity is part of our life. All living organisms exhibit in some way or another a form of rhythmicity. Some of these rhythms have a geophysical counterpart in the environment. These are the so-called biological rhythms. This paper gives an introduction to the circadian rhythms, i.e., those rhythms with a period length of about one day. General characteristics and properties are described as well as the control and disturbances of these rhythms. PMID- 8731334 TI - Studying the snail's clock at better than a snail's pace. AB - The eye of Bulla gouldiana, the cloudy bubble snail, contains a circadian pacemaker that times a nocturnal locomotor rhythm. The eye expresses a circadian rhythm in spontaneous impulse activity that is generated within individual neurons at the base of the retina. There are approximately 100 of these "clock neurons" that are electrically interconnected. The retinal clock can be synchronized by light cycles. Synchronization involves membrane depolarization and calcium influx into pacemaker neurons. While rhythm expression involves an ionic calcium flux, and rhythm expression appears to be mediated by changes in potassium conductance, circadian rhythm generation does not appear to rely on transmembrane fluxes. Rather, the molecular events of transcription and translation appear to be critical players in generating the near twenty-four-hour rhythm in optic nerve activity. PMID- 8731335 TI - Pacemaker interactions in the mammalian circadian system. AB - Circadian rhythms in mammals are generated by pacemaker cells located in the suprachiasmatic nucleus (SCN) of the anterior hypothalamus. The identity of these cells, however, is not known, and little information exists regarding the mechanisms by which they communicate with each other and with the organism. Nonetheless, pacemaker interactions must occur to produce single, coherent rhythms of behavior and physiology. Recently it has become possible to observe the result of these interactions using circadian chimeras, animals with two clocks with distinct periods, that have been produced by SCN transplantation. Using the tau mutation in golden hamsters, chimeras expressing two circadian rhythms of behavior simultaneously were created. The two rhythms exhibited complex interactions including cases of relative coordination. This basic result indicates that pacemaker interactions are rhythmic and phase dependent. Further analysis should help to elucidate the nature of the coupling signal and the identity of the pacemaker cells. PMID- 8731336 TI - Circadian rhythms in the neonate and in old age: what do they tell us about the development and decay of the body clock in humans? AB - This overview describes changes in human circadian rhythms in the neonate and in aged subjects. Since a measured circadian rhythm is derived from an endogenous (clock-driven) component and an exogenous component (due to rhythms in the environment and lifestyle), this account attempts to separate these possibilities in any particular case. Experimental methods for distinguishing between these components are described, and it is concluded that the data are rarely clear enough for such distinctions in mechanism to be made with confidence. Nevertheless, the evidence to date suggests that changes to the endogenous and exogenous components during both the development and decay of circadian rhythms are involved. This being the case, and accepting that poorly developed circadian rhythms are often associated with poor general health and development, methods to strengthen circadian rhythms are described. PMID- 8731337 TI - Recent progress in understanding the temporal behavior of unicellular organisms. AB - This survey summarizes the findings concerning endogenous oscillations of three unicellular organisms: the dinophyte Gonyaulax polyedra, the ciliate Tetrahymena thermophila and the euglenophyte Euglena gracilis. All of them behave rhythmically and show the common features of zeitgeber action, differential sensitivity and temperature compensation; however, they exhibit some species specific peculiarities that make each of them suitable for addressing particular chronobiological questions. Although ultradian rhythms have been described for Tetrahymena thermophila and Euglena gracilis, they appear under different conditions: in the first case, a modulation of the period in relation to the concentration of nutrients is observed, whereas Euglena oscillates in an ultradian and circadian fashion simultaneously. Transitions between periodic and aperiodic states can be induced in Euglena gracilis and Gonyaulax polyedra: Euglena gracilis can enter an aperiodic state after repeated exposure to short light pulses (up to 10 sec) given at intervals of 40 min or less, whereas in Gonyaulax polyedra the circadian oscillator is arrested at temperatures below 12 degrees C. In the arrhythmic state, the oscillator might be driven into singularity within the phase space of a limit cycle attractor; re-initiation from the holding point occurs by transition to a relatively precisely defined new phase. Photoperiodism as another important chronobiological phenomenon can be studied in Gonyaulax polyedra: cells enter the dormant stage of an asexual cyst under short days and a temperature below 16 degrees C. This response can be mimicked by 5-methoxylated indoleamines such as melatonin and 5 methoxytryptamine, which are synthesized by this organism. Melatonin concentration exhibit an endogenous circadian rhythm characterized by a rapid increase shortly after the onset of darkness. Encystment, as induced by indoleamines, is associated with stimulations of bioluminescence. The coupling of the two processes involves, as a common element, the release of protons from an acidic vacuole. PMID- 8731338 TI - Several clocks may simplify the circadian system of Gonyaulax. AB - The circadian system of Gonyaulax involves at least two pacemaking oscillators. These oscillators have been shown to be distinct at the physiological level by an examination of their rhythmic periods under different conditions, their phase response to pulses of light and darkness and their ability to compensate the rhythms for changes in temperature. It is also known that Gonyaulax can restrict synthesis of different proteins to at least three different times of day. We suggest that the two known oscillators may each control synthesis of a differently timed class of proteins. PMID- 8731339 TI - The effect of light on the biosynthesis of beta-carotene and superoxide dismutase activity in the photosynthetic alga Gonyaulax polyedra. AB - Daily oscillations of both beta-carotene and superoxide dismutase (SOD) activity are related to the intracellular control of reactive oxygen species (ROS). It is well established that ROS are present in all aerobic cells. We studied the marine dinoflagellate Gonyaulax polyedra which has been extensively used as a model to understand the biological clock at the molecular level. beta-Carotene, besides suppressing singlet molecular oxygen (1O2), may act as a photoreceptor pigment in many photosynthetic cells. The levels of beta-carotene during the day phase were shown to be twice as high as during the night phase. The dose-response curve for light-induced carotenoid synthesis was linear for up to 45 min of light exposure, after which night phase cells contained the same levels of beta-carotene as day phase cells. Cells exposed to light pulses at different times during the dark period displayed the highest beta-carotene induction in the middle of the night. SOD activity of cell-free extracts of G. polyedra was three to four times higher during the day. This rhythm continued in cells kept in constant light, indicating that the regulation can be attributed to the cellular circadian clock. No denaturing polyacrylamide gels revealed the presence of several SOD isoenzymes in G. polyedra, including CuZnSOD and MnSOD. Furthermore, G. polyedra SOD cross reacts with a polyclonal antibody raised against SOD. In addition to being gene regulated by ROS concentration, G. polyedra SOD expression seems also to be under the control of the biological clock. PMID- 8731340 TI - Imaging oscillations in Gonyaulax: a chloroplast rhythm of nitrate reductase visualized by immunocytochemistry. AB - Gonyaulax polyedra is a unicellular marine photosynthetic dinoflagellate known to display numerous circadian rhythms, including bioluminescence, motility, cell division and several chloroplast-related rhythms. Due to this, Gonyaulax has become a widely used model organism for studying the cellular biological clock. In this work we describe another rhythm for Gonyaulax cells also associated with the cell's chloroplasts, a rhythm in localization of the enzyme nitrate reductase (NR). A polyclonal antibody was raised against NR purified from G. polyedra cells and used as a probe in immunogold labelling experiments on cell thin sections, comparing day- and night-phase cells. The enzyme localizes to chloroplasts in day phase cells, while the enzyme is active, and is largely absent in night-phase cells. Counts of gold particle distribution in day- versus night-phase cells show an approximate three-fold increase in enzyme labelling in day-phase plastids. These results closely approximate the four-fold differences shown for NR activity between day and night Gonyaulax cells by biochemical studies. We conclude from the diurnal difference in labelling that NR is localized in Gonyaulax chloroplasts during the day phase and is absent (broken down) in night-phase cells. Thus NR in Gonyaulax is compartmentalized in the chloroplasts and is therefore subject to similar circadian control mechanisms exhibited for other plastid rhythms. PMID- 8731341 TI - Chronobiology of indoleamines in the dinoflagellate Gonyaulax polyedra: metabolism and effects related to circadian rhythmicity and photoperiodism. AB - The marine bioluminescent dinoflagellate Gonyaulax polyedra is capable of producing various indoleamines. The first enzyme in their formation, tryptophan hydroxylase, exhibits a high-amplitude circadian rhythm with a maximum during photophase. Hydroxyindole-O-methyltransferase shows a biphasic pattern with a major maximum during scotophase. 5-Methoxylated indoleamines, such as melatonin and 5-methoxytryptamine, peak at the beginning and in the second half of scotophase, respectively. A drop in temperature from 20 to 15 degrees C leads to dramatic increases of melatonin, up to more than 50 ng/mg protein. This effect may explain why a lower temperature sensitizes this organism to photoperiodic, indoleamine-mediated induction of asexual cysts. Melatonin can be catabolized either enzymatically or non-enzymatically. The non-enzymatic pathway involves free radicals, e.g., photooxidant cation radicals, and leads to the formation of N1-acetyl-N2- formyl-5-methoxykynuramine. Enzymatic catabolism comprises deacetylation to 5-methoxytryptamine and formation of 5-methoxytryptophol. 5 Methoxytryptamine represents a key substance acting as a stimulator of bioluminescence and a mediator of the encystment response. It opens proton channels in the membrane of an intracellular acidic vacuole system which is loaded by the action of a V-type ATPase, as shown by experiments using bafilomycin A1. PMID- 8731342 TI - Becoming circadian--A one-year study of the development of the sleep-wake cycle in children. AB - Data on three children's sleep-wake behavior during the first year of life submitted to spectral analysis reveal the presence of several frequencies in the circadian and ultradian ranges. The changes in the resulting spectra as age progresses show both an increase in overall rhythmicity, i.e., more data are explained by regular oscillations, and irregular changes in the circadian component. This circadian component shows an increase in power which appears at different ages among the individuals studied. PMID- 8731343 TI - Let there be light: signal transduction in a mammalian circadian system. AB - Mammalian circadian rhythms are controlled by a biological clock located in the hypothalamic suprachiasmatic nuclei (SCN). This clock is entrained by light through a retinohypothalamic pathway that interacts with the SCN through glutamate neurotransmission. Light pulses during the subjective night induce phase shifts of behavioral rhythms, and also trigger intracellular changes such as the expression of immediate-early genes and activation of transcription factors. In this review, we present a model of the signal transduction pathway leading to photic synchronization of the circadian clock, including the activity of specific second messenger systems, gene expression, and interaction between potential agents capable of producing phase shifts. PMID- 8731344 TI - Control of environmental variables in a field study using a chronobiological protocol. AB - Field work in chronobiology usually is based on observations which differ from those in laboratory studies which are experimental. Non-invasive protocols are recommended in the field in order not to introduce additional variables. We propose an alternative strategy here. The effect of environmental cycles on biological rhythms can be estimated when the observations are made in more than one area. The areas under study must have at least one characteristic in common which is a guarantee that the limits of oscillation of an environmental cycle will be similar. This proposal has been tested using the biological system consisting of an Onagraceae plant, Ludwigia elegans, and its bee visitors, which is found in two localities at the same latitude, but at different altitudes. Under the same photoperiod, but with the mean temperature ranging within known limits, it was possible to determine temporal characteristics of the system in the field by examining the effects of the light/dark and temperature daily cycles. PMID- 8731345 TI - Context and Pavlovian conditioning. AB - Procedurally, learning has to occur in a context. Several lines of evidence suggest that contextual stimuli actively affect learning and expression of the conditional response. The experimental context can become associated with the unconditional stimulus (US), especially when the US is presented in a context in the absence of a discrete conditional stimulus (CS). Moreover, context can modulate CS-US associations. Finally, it appears that context can become associated with the CS when it is presented before the CS-US training. The purpose of the present paper is to review some of the relevant literature that considers the context as an important feature of Pavlovian conditioning and to discuss some of the main learning theories that incorporate the context into their theoretical framework. The paper starts by mentioning historical positions that considered context an important variable in conditioning and then describes how the approach to contextual conditioning changed with the modern study of Pavlovian conditioning. Various forms of measurement of context conditioning are presented and the associative strength attached to context in several experimental paradigms is examined. The possible functions that context may acquire during conditioning are pointed out and related to major learning theories. Moreover, the effect of certain neurological manipulations on context conditioning is presented and these results are discussed in terms of possible functions that the context might acquire during Pavlovian conditioning. It is concluded that contextual stimuli acquire different functions during normal conditioning. A procedure in which animals are exposed to an aversive US immediately after they are placed in the experimental context is suggested as a useful control for the study of context conditioning. PMID- 8731346 TI - The reaction between ABTS radical cation and antioxidants and its use to evaluate the antioxidant status of serum samples. AB - The 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical cation can be generated by incubation of ABTS and 2,2'-azo-bis(2- amidinopropane) at 45 degrees C. The ABTS radical cation is stable for several minutes at room temperature and reacts quantitatively and instantaneously with several antioxidants, such as Trolox, ascorbic acid, uric acid, cysteine, glutathione and bilirubin. In contrast, the ABTS radical cation reacts slowly with albumin. When serum is added to a solution of the ABTS radical cation, the bleaching of the radical follows biphasic kinetics, with a fast decay followed by a slow decay that takes place within several minutes. The fast decay is primarily due to uric acid, while the slow decay is related to the protein content of the sample. We propose that this procedure can provide an independent and simultaneous evaluation of the low molecular weight and protein antioxidants present in biological samples such as serum. PMID- 8731347 TI - An alternative column chromatographic process for the production of human albumin. AB - We have modified a standard column chromatography method for the preparation of albumin from human plasma. The proposed method utilizes Sephadex G-25, euglobulin precipitation, DEAE-Sepharose, ethanol heat treatment and Sephacryl S-200 HR. The procedure differs from that normally used by the introduction of Schneider's ethanol/heat treatment step and the elimination of a CM-Sepharose step, after the DEAE-Sepharose step. The proposed method was used to produce 15 batches of albumin, 100 liters of plasma per batch. Purity of more than 99% was obtained at an average yield of 26.5 g/l plasma for the albumin produced. All batches presented 99.2 to 99.9% monomer and 0.1 to 0.8% dimer, with no larger polymers detected. The utilization of the final gel filtration step eliminated highly polymerized albumin usually obtained during the process. The advantages of the proposed method are reduction in the overall process time from 6 to 5 days, elimination of the CM-Sepharose step and the reduction of 6 to 4 columns in series for the Sephacryl S-200 HR step. PMID- 8731348 TI - A specific and highly sensitive time-resolved fluoroimmunoassay for human proinsulin. AB - We describe a time-resolved fluoroimmunoassay specific for human proinsulin using a combination of two high-affinity monoclonal antibodies, one against insulin and the other specific for intact proinsulin and for split 65-66 and des 64-65 proinsulin forms. The assay employs only 200 microl of serum, with a detection limit of 0.1 pmol/l. The intra-assay variation coefficient was less than 3% between 3 and 1000 pmol/l. There was 0% cross-reaction with insulin, C-peptide, split 32-33 and des 31-32 proinsulin. Serum concentration of proinsulin was analyzed in 50 subjects during an oral glucose tolerance test (10 non-obese control, 10 obese controls, 10 subjects with impaired glucose tolerance, 10 patients with type II diabetes mellitus (DM) and fasting blood glucose (FBG) < 140 mg/dl, and 10 patients with type II DM and FBG > 150 mg/dl). Mean fasting serum proinsulin levels measured by this assay in non-obese controls (0.84 - 0.90 pmol/l; 0.1-2.4 pmol/l) were lower than the results reported by other investigators. There was an increase of proinsulin related to obesity and increased glucose levels, suggesting that proinsulin levels increase with insulin resistance. PMID- 8731349 TI - Transfer of IgG subclasses across placenta in term and preterm newborns. AB - In order to study placental transfer of IgG subclasses, paired blood samples were collected from mothers and umbilical cord of preterm (N = 69) and full-term (N = 68) newborns. The full-term group was further divided into 3 subgroups: appropriate for gestational age (AGA, N = 43), large for gestational age (LGA, N = 13) and small for gestational age (SGA, N = 12), according to birth weight. IgG subclasses (IgG1, IgG2, IgG3 and IgG4) were measured by the single radial immunodiffusion technique using monoclonal antibodies. IgG1 and IgG3 newborn subclass concentrations (10.17 and 0.57 g/l, respectively) increased with increasing gestational age and reached maternal levels (IgG1 = 8.86; IgG3 = 0.67 g/l) during the 37th week of pregnancy. Low levels of these subclasses were found in premature newborns. IgG2 from newborns were always lower than maternal levels (P < 0.05). LGA and SGA newborns had equivalent levels of IgG1 and IgG2 compared with AGA. SGA newborns had higher levels of IgG3 and lower levels of IgG4 than LGA and AGA newborns. PMID- 8731350 TI - Effect of Phoneutria nigriventer spider venom on gastric emptying in rats. AB - The effect of Phoneutria nigriventer spider venom (PNV) on the gastric emptying of liquids was studied in 240 young adult Wistar rats (2-3 months of age) divided into subgroups of 8 animals each. The study was performed in 3 stages. Initially, PNV was injected into rats at doses of 0.19, 0.38 or 0.76 mg/kg and the effect on gastric emptying was assessed 30 min later. In the second stage, a time-course study was performed by injecting 0.76 mg PNV/kg and measuring the effect on gastric emptying 15, 60 and 120 min post-venom. In the last stage, in order to investigate the possible mechanisms of PNV influence on gastric emptying, one group of rats underwent subdiaphragmatic vagotomy and then received 0.76 mg PNV/kg while three other groups were pretreated iv with either prazosin (0.4 mg/kg), domperidone (1.0 mg/kg) or propranolol (0.6 mg/kg) and then given 0.38 or 0.76 mg PNV/kg. In this last stage, gastric retention was measured 30 min post venom. Each animal received a saline test meal solution containing phenol red as a marker (60 micrograms/ml). Ten min after administering the test meal by gavage, gastric retention was determined by measuring the residual test meal marker concentration and the animals were sacrified. PNV (0.76 mg/kg) provoked a significant delay in gastric emptying of liquids 15, 30 and 60 min after its administration. Propranolol partially interfered with gastric emptying in rats that had received 0.38 and 0.76 mg PNV/kg. Vagotomy and pretreatment of the rats with prazosin and domperidone had no effect. We conclude that the delay in the liquid gastric emptying observed in severely envenomed rats was probably due, at least in part, to a venom-stimulated release of catecholamines which inhibited gastric motility by activating smooth muscle beta-adrenergic receptors. PMID- 8731351 TI - Differential effects of protein restriction on gamma-glutamyltranspeptidase [EC 2.3.2.2] activity in young and mature rats. AB - Gamma-glutamyltranspeptidase (GGT, EC 2.3.2.2) activity was determined in the plasma and liver of 40 young (50 days old) and 40 mature (300 days old) male Wistar rats, after a protein restriction period of 28 days. Casein protein levels used were: 1%, 3%, 5%, 7% and 28% (control). Weanling rats submitted to protein free diet and adult rats submitted to that and other low-protein diets (1%, 3% or 5% casein) presented weight reductions (-0.38 +/- 0.07 g/day and -0.98 +/- 0.19 g/day, respectively). Only in young animals did these weight reductions parallel those of food consumption (37 +/- 12% of control), plasma protein (52 +/- 11% of control), plasma albumin (70 +/- 12% of control), hepatic RNA (68 +/- 7% of control) and protein (71 +/- 8% of control). A marked effect of the protein restriction, increasing the GGT activity, was also observed only in young rats. A significant (P < 0.05) rise was promoted by the protein-free diet in the plasma GGT (2.83 +/- 1.39 vs 0.69 +/- 0.50 mU/ml for control) and by both the protein free and 3% casein diets, in the liver GGT (respectively, 16.00 +/- 6.72 and 7.75 +/- 3.49 vs 0.94 +/- 0.57 U/g protein for control). The different results obtained for young in relation to mature animals could be explained by the reduction of both protein and sulfur-containing amino acid requirements with aging. PMID- 8731352 TI - Effect of isoprinosine on rotavirus replication in vitro. AB - Isoprinosine (IPS) is a synthetic drug whose antiviral effect on rotavirus replication in vitro has been characterized in terms of the decrease in metachromasia after acridine orange staining. The present study describes the effect of IPS on the synthesis of viral RNA in vitro. MA-104 cell cultures infected with simian rotavirus strain SA-11 were incubated with zero, 250, 500 and 1,000 micrograms/ml IPS and 22, 24, 48, 52, 72 and 76 h after infection the cultures were submitted to a 1-h starvation period, followed by a 2-h pulse with 10 microCi/ml of [3H]-uridine. The homogenates of virus-infected cultures treated or not with IPS were submitted to phenol/chloroform extraction followed by polyacrylamide gel electrophoresis. The amount of radioactivity in viral RNA eluted from the gel strips was determined. Inhibition of viral RNA synthesis was highest at the IPS concentration of 1,000 micrograms/ml at 72 h after infection, corresponding to 78% inhibition. Although the results obtained in vitro suggest that IPS may be useful for the treatment of rotavirus infection, an in vivo demonstration of its efficacy is needed. PMID- 8731353 TI - Study of the embryotoxic effects of an extract of rosemary (Rosmarinus officinalis L.). AB - Extracts of rosemary, Rosmarinus officinalis L., have been used in folk medicine as a diuretic, an emenagogue, an antispasmodic and its aqueous extract does not present toxicity to man, presenting, however, abortive effects. In order to evaluate if this plant induces abortion and/or interferes with the normal development of the concepts, doses of 26 mg of a 30% (w/v) R. officinalis aqueous extract (13 mg solids/ml) made with leaves, flowers and stem were administered daily by gavage during two different periods of Wistar rat pregnancy. One group of animals (N = 12) received the extract from days 1 to 6 of pregnancy (preimplantation period) and another group (N = 14) received the same extract from days 6 to 15 of pregnancy (organogenic period). Control groups (N = 12) received saline in the same volume and during the same periods as their respective experimental groups. The animals were sacrificed at term. The treatment of the dams during either the preimplantation or the organogenic period did not cause significant changes in the postimplantation loss or in the number of anomalies or malformations of the term fetuses, which also showed a similar degree of development when compared with the respective controls. The percent of preimplantation loss in the group treated before embryo implantation increased, although the difference was not significant compared to the control. This result suggests that rosemary extract may present an anti-implantation effect without interfering with the normal development of the concept after implantation. PMID- 8731354 TI - Terminal maturation of resting B cells induced by macrophage factors. AB - Mouse splenic macrophages from BALB/c nude mice (purified by plastic adherence) or cloned macrophage hybridomas stimulated with jacalin (12.5 micrograms/ml), a D Gal binding lectin, produce one or more B-cell stimulatory factors which cause splenic B cells from BALB/c or C3H/HeJ mice to secrete immunoglobulin in a polyclonal manner as detected by reverse protein A plaque assays. Jacalin stimulated macrophage supernatants (JacSup) activate both normal and Percoll gradient-purified small high-density (resting) B cells. Supernatants from total or resting BALB/c spleen cells cultured for 7 days in the presence of JacSup (derived from splenic BALB/c nude mice macrophages) were assayed for immunoglobulin isotypes by ELISA. Resting B cells produce only IgG3 and IgM, whereas total B cells secrete IgG3 and IgM as well as IgG1, IgG2a, IgG2b and IgA. Resting and total B cells from BALB/c nude mice are also stimulated by macrophage supernatants to secrete immunoglobulin, thus indicating that this activity is likely to be T cell independent. Moreover, jacalin-stimulated macrophage supernatants did not induce spleen cells or purified B cells to proliferate. Fractionation of factor-rich supernatants on a Sephacryl S-200 column revealed that the factor activity is located in fractions corresponding to a molecular mass of 25-27 kDa. Taken together, these results suggest that upon the action of a macrophage factor(s) resting B cells undergo terminal differentiation without proliferation in the absence of T cells. PMID- 8731355 TI - An in vitro model to study the effect of antibodies and cell receptor analogues on meningococcal adherence to human oroepithelial cells. AB - We have investigated different experimental schedules to achieve adherence of Neisseria meningitidis group B to cultured and buccal epithelial cells (BEC) and the effect of antibodies and receptor analogues on bacterial adherence. No adherence of meningococcus was observed when HeLa, HEp-2 or KB cells were used, but high rates of adherence to BEC occurred. The effect of antibodies on bacterial adherence was studied in assays carried out in the presence of saliva and serum collected from convalescing children with meningococcal meningitis and children vaccinated with VAMENGOC B-C. Both saliva and serum from the convalescent patients inhibited the adherence of meningococci, but saliva and serum from vaccinated children did not, corroborating our previous data of a poor antibody response induced by this vaccine. Human colostrum did not affect meningococcal adherence despite the presence of antibodies to N. meningitidis detected by ELISA. Inhibition of adherence by sera from an immunized horse, rabbits and mice, as well as by cell receptor analogues (outer-membrane complex and purified polysaccharide C), was observed. Our results show that up to now BEC continue to be the best cells to study meningococcal adherence and the effect of adherence inhibitors. PMID- 8731356 TI - Effect of a non-peptide angiotensin receptor antagonist on water intake caused by centrally administered carbachol in the rat. AB - Angiotensin II (ANG II) administered centrally produces drinking by acting on subtype 1 ANG II (AT1) receptors. Carbachol, a cholinergic receptor agonist, also induces drinking behavior by a central action. In the present study we determined whether the response to carbachol also involves AT1 receptors. Male Holtzman rats (250-300 g) with stainless steel cannula implanted into the lateral ventricle (LV) were used. Water intake after injection of 0.15 M NaCl (1.0 microliter) into the LV was 0.2 +/- 0.01 ml/h (N = 8). The AT1 receptor antagonist DUP-753 (50 nmol/microliters) injected into the LV reduced water intake induced by ANG II (10 nmol/microliters) from 9.2 +/- 1.4 to 0.4 +/- 0.1 ml/h (N = 8), and water intake induced by carbachol (2 nmol/microliters) from 9.8 +/- 1.4 ml/h to 3.7 +/- 0.8 ml/h (N = 8). These results suggest that AT1 receptors play a role in the drinking behavior observed after central cholinergic stimulation in rats. PMID- 8731357 TI - N-acetylaspartylglutamate acetoxymethyl triester (NAAG.AM) as a tool for loading the neuropeptides NAAG and succinimidyl-NAAG into intact cells: effect on [3H] dopamine exocytosis. AB - Although N-acetylaspartylglutamate (NAAG) is one of the neuropeptides found in highest concentrations in the mammalian central nervous system, its functional role in neuronal signaling has not been definitively established. In some neuronal populations, NAAG is concentrated in nerve terminals and thus, it may play a role in the cytoplasmic events underlying neurotransmitter exocytosis. In the present study we have validated the use of the synthetic derivative NAAG acetoxymethyl triester (NAAG.AM) as a tool to increase the intracellular levels of the peptide and assessed the ability of NAAG to regulate [3H]-dopamine ([3H] DA) secretion in PC12 cells. Enzymatic degradation of NAAG.AM by nonspecific brain esterases resulted in the progressive formation of NAAG and succinimidyl NAAG (Asu-NAAG). However, only 8% of NAAG.AM was converted to NAAG. Significant amounts of NAAG (1 nmol/mg protein) were demonstrable in cultures of the neuroblastoma cell line N2A following incubation with NAAG.AM for 2 h, with the concentration of (Asu)-NAAG being at least 100-fold higher. The pheochromocytoma cell line PC12 was used to assess the influence of loaded NAAG derivatives on [3H]-DA exocytosis. Incubation with 0.1-1 mM NAAG.AM did not affect the basal efflux or total content of [3H]-DA. However, it induced a dose-dependent decrease of [3H]-DA secretion in response to 56 mM KCl depolarization reaching an inhibition of 49% with 1 mM NAAG.AM. In contrast, NAAG.AM did not affect secretion induced by the calcium ionophore A23187 (100 microM). The present study validates the use of NAAG.AM as a tool to load NAAG derivatives into intact cells and provides preliminary evidence for an intracellular role of the peptide. PMID- 8731358 TI - Mild depression levels alter self-perceptions of future but not the recall of verbal information in elderly inpatients. AB - In order to determine the correlation of levels of symptoms of depression and rate of forgetting and perception of the future, a total of 68 elderly inpatients without Major Depression admitted to a general hospital were evaluated by: 1) the Montgomery-Asberg Depression Rating Scale (MADRS), 2) the Mini-Mental State Examination (MMSE), 3) a questionnaire on future self-perceptions (FSPQ), and 4) a test on the recall of verbal information to estimate the rate of forgetting. They were grouped according to the clinical prognosis of their disease (good, N = 48, 25 women, 23 men, age mean +/- SD, 68 +/- 6.64; poor, N = 20, 10 women, 10 men, age mean +/- SD, 69 +/- 6.68) which correlates with morbidity-mortality rates (low/high). There was no relationship between mild levels of signs and symptoms of depression and increased forgetting. However, levels of depression were negatively correlated to the score of future perceptions (B = -0.18, beta = 0.29, P = 0.032). Patients with diseases with good prognosis did not present different levels of depression, rates of forgetting or future expectations from those of patients with poor prognosis (high mortality rates). However, individuals with negative FSPQ scores showed significantly higher MADRS scores, independent of the type of disease. These data suggest that the modifications in the processing of information related to the future are present in clinical patients without Major Depression but they occur within a small range of very mild signs and symptoms of depression. PMID- 8731359 TI - The lysine-1 analog of guanylin induces intestinal secretion and natriuresis in the isolated perfused kidney. AB - Guanylin is an endogenous peptide synthesized by several mammalian species that mimics the effects of a thermostable enterotoxin of Escherichia coli (STa: NTFYCCELCCNPACAGCY) in the gut. We have cloned a lysine-1 derivative of rat guanylin (Lys-1-NTCEICAYAACTGC) and tested its effects on ileal tissue membranes in Ussing chambers and in the isolated perfused rat kidney. Rabbit ileal mucosa membranes were mounted into a Ussing chamber and the effects of Lys-1 guanylin (Lys-1 G) and STa enterotoxin peptide on chloride secretion were determined by changes in short-circuit current (Isc). Lys-1 G (10 to 100 nM) showed a dose dependent effect on chloride secretion with a maximal response estimated to be 52 microA/cm2. Lys-1 G mimics the effect of STa peptide, but the enterotoxin elicited a greater maximal effect of 120 microA/cm2 (P < 0.01). Lys-1 G (2.5 micrograms/ml) promoted an increase in both urine flow (from 0.13 +/- 0.07 to 0.40 +/- 0.01 ml g-1 min-1, N = 4; P < 0.05) and glomerular filtration rate (from 0.68 +/- 0.02 to 0.85 +/- 0.00 ml g-1 min-1, N = 4; P < 0.01) in the isolated perfused kidney and a reduction of the fractional reabsorption of sodium (from 76.0 +/- 0.03 to 59.5 +/- 0.85%, N = 4; P < 0.01). These maximal effects were accompanied by intense natriuretic effect observed 30 and 60 min after drug administration. The Lys-1 G analog similar to STa enterotoxin elicited intestinal chloride secretion and a natriuretic effect. These data demonstrate that the cloned peptide analog retains the biological activity of the native hormone and presents activity similar to STa. The properties of Lys-1 G resemble those of a factor formed during perfusion of the hypoxic rabbit kidney and named by us factor natriureticus similis (FNS). PMID- 8731360 TI - Altered gonadal hormone level and constant light-induced stress interfere with the response of the adrenal medulla to oxytocin. AB - The effect of oxytocin (0.25 IU/100 g per day) on the adrenal medulla was examined in intact, intact estrogen-treated, castrated and castrated testosterone treated adult male Wistar rats. Stereological analysis of the gland (N = 5 rats per group) revealed that in intact animals the number of chromaffin cells (x10(3)) was significantly increased after 3-day (saline: 467.6 +/- 27.4; oxytocin: 567.6 +/- 28.9) or 7-day (saline: 486.2 +/- 39.1; oxytocin: 618.7 +/- 36.8) oxytocin administration. During 7 days of recovery after the 7-day treatment, the chromaffin cell number returned to the control level (saline: 491.4 +/- 12.6; oxytocin: 554.4 +/- 28.7). The effect of oxytocin on chromaffin cell number was also observed in rats simultaneously injected with estradiol (0.3 micrograms/100 g per day) for 10 days (estradiol: 454.3 +/- 32.8; estradiol+oxytocin: 576.1 +/- 25.0), as well as in 10-day castrated rats (saline: 594.7 +/- 22.7; oxytocin: 765.3 +/- 33.1). Testosterone replacement (0.6 mg/100 g per day) abolished the medullary response to oxytocin (testosterone+saline: 528.5 +/- 24.7; testosterone+oxytocin: 620.8 +/- 56.0). There was a 20% rise in adrenal dopamine content (from 0.236 +/- 0.015 to 0.283 +/- 0.015 microgram per pair of glands; N = 9-12) in intact rats injected with oxytocin for 3 days. Oxytocin had no effect on any of the catecholamine levels in adrenal glands of rats exposed to stress induced by constant lighting. The present data indicate that the proliferative response of chromaffin tissue to oxytocin depends on the gonadal hormone level and the basal activity of the adrenal medulla. PMID- 8731361 TI - Acute respiratory infections in children living in two low income communities of Rio de Janeiro, Brazil. AB - Community studies of non-hospitalized children are essential to obtain a more thorough understanding of acute respiratory infections (ARI) and provide important information for public health authorities. This study identified a total ARI incidence rate (IR) of 4.5 per 100 child-weeks at risk and 0.78 for lower respiratory tract infections (LRI). Disease duration averaged less than one week and produced a total time ill with ARI of 5.8% and for LRI 1.2%. No clear seasonal variation was observed, the sex-specific IR showed a higher proportion of boys becoming ill with ARI and LRI and the peak age-specific IR occurred in infants of 6-11 months. Correlation with risk factors of the child (breastfeeding, vaccination, diarrheal disease, undernourishment) and the environment (crowding, living conditions, maternal age and education) showed marginal increases in the rate ratios, making it difficult to propose clear-cuts targets for action to lower the ARI and LRI morbidity. The importance of an integral maternal-child health care program and public education in the early recognition of LRI is discussed. PMID- 8731362 TI - Giardia survey in live-trapped small domestic and wild mammals in four regions in the southwest region of the state of Sao Paulo, Brazil. AB - For the first time, a survey on Giardia in live-trapped small domestic and wild mammals was performed in four regions of State of the Sao Paulo, Brazil, with special attention to the parasitism of Rattus rattus rattus by Giardia. This species was found infected in all studied sites: Botucatu (15.4%), Conchas (28.5%), Itaporanga (38.7%) and Sao Roque (100%). Two new hosts and their frequency of infection were described for Giardia in Nectomys squamipes, an aquatic rodent and in Bolomys lasiurus, a forest rodent (100% and 14.3%, respectively). Both G. muris and G. duodenalis groups were found in scrapings of intestinal mucosa of those rodents. Mixed infection was observed in some animals. It is important to emphasize the infection by G. duodenalis in the black rat as this species lives as a comensal with man and in N. squamipes as it may contaminate small streams used for domestic consumption. Therefore, further investigation will be necessary to elucidate the potencial of these rodents to act as reservoirs of Giardia for man. PMID- 8731363 TI - Interaction between Didelphis albiventris and Triatoma infestans in relation to Trypanosoma cruzi transmission. AB - This paper attempts to prove if a high Trypanosoma cruzi prevalence of opossums might be reached with few potential infective contacts. One non-infected Didelphis albiventris to T. cruzi and 10 infected nymphs of Triatoma infestans were left together during 23 hr in a device that simulated a natural opossum burrow. Twenty-six replicates were performed using marsupials and triatomines only once. Potentially infective contacts occurred in all the trials. From the 26 opossums used in trials, 54% did not eat any bug. Of the 260 bugs used, 21% were predated. In the 25 trials involving 205 surviving bugs, 36% of them did not feed. In 15/25 cases, > or = 60% of the triatomines were able to feed. The parasitological follow-up of 24 opossums showed that among 10 that had eaten bugs, 4 turned out infected and among the 14 that had not predate, 3 (21%) became positive. In sum, 7/24 (29%) of the marsupials acquired the infection after the experiment. This infection rate was similar to the prevalences found for the opossum population of Santiago del Estero, Argentina, suggesting that the prevalences observed in the field might be reached if each marsupial would encounter infected bugs just once in its lifetime. PMID- 8731364 TI - V3 peptide binding pattern and HIV-1 transmission route in Rio de Janeiro. AB - To characterize antibody binding to a panel of V3 loop peptides representing diverse HIV-1 neutralization epitopes, 149 HIV-1 infected individuals from Rio de Janeiro (RJ) were investigated. Results were analyzed with respect to risk factors for infection and other epidemiological and clinical data. Peptide reactivity was not associated with sex, clinical status, CD4 counts, antigenemia or beta 2-microglobulin serum level. A segregation of peptide reactivity according to route of infection was encountered. This finding suggests that more then one viral strain may be circulating in RJ, in subjects with different risk factors for HIV-1 infection. An investigation of prevalent HIV-1 genotypes, serotypes and immunotypes may be of importance for the design and selection of potential vaccines to be used in Brazil as well as for the selection of populations to be included in future vaccine efficacy trials. PMID- 8731365 TI - Search for Trypanosoma cruzi in the anal glands of wild Didelphis albiventris from Santiago del Estero, Argentina. PMID- 8731366 TI - Angiostrongylus costaricensis life cycle: a new proposal. PMID- 8731367 TI - Prevalence of anti-Taenia solium antibodies in sera from outpatients in an Andean region of Ecuador. AB - Sera from 9,254 individuals that presented at one of three outpatient clinics in Quito, Ecuador were assayed by indirect hemagglutination for the presence of antibodies reactive with antigens from Taenia solium cysts. Immunoblot analysis of 81 selected sera with IHA titers ranging from 0 to 1,028 showed that a titer of > or = 32 was suggestive of exposure to the parasite. Nine percent (9%) of the 9,254 patients had titers of 32 or greater. Of 3,503 sera from one clinic, which included sera from food handlers undergoing yearly physicals, 390 (11%) were positive. In addition, a correlation with age was seen in some, but not all, populations. In situations where age-related effects were noted, the highest incidence was seen in the youngest (0-20 years) and in the oldest (51-60 years) group. Thus, a resurgence of infection after a period of lower prevalence may be developing. Overall, this study shows that cysticercosis is relatively common and potentially a serious health problem in this region. PMID- 8731368 TI - Identification of species related to Anopheles (Nyssorhynchus) albitarsis by random amplified polymorphic DNA-polymerase chain reaction (Diptera: Culicidae). AB - Species-specific Random Amplified Polymorphic DNA-Polymerase Chain Reaction (RAPD PCR) markers were used to identify four species related to Anopheles (Nyssorhynchus) albitarsis Lynch-Arribalzaga from 12 sites in Brazil and 4 in Venezuela. In a previous study (Wilkerson et al. 1995), which included sites in Paraguay and Argentina, these four species were designated "A", "B", "C" and "D". It was hypothesized that species A is An. (Nys.) albitarsis, species B is undescribed, species C is An. (Nys.) marajoara Galvao and Damasceno and species D is An. (Nys.) deaneorum Rosa-Freitas. Species D, previously characterized by RAPD PCR from a small sample from northern Argentina and southern Brazil, is reported here from the type locality of An. (Nys.) deaneorum, Guajara-Mirim, State of Rondonia, Brazil. Species C and D were found by RAPD-PCR to be sympatric at Costa Marques, State of Rondonia, Brazil. Species A and C have yet to be encountered at the same locality. The RAPD markers for species C were found to be conserved over 4,620 km; from Iguape, State of Sao Paulo, Brazil to Rio Socuavo, State of Zulia, Venezuela. RAPD-PCR was determined to be an effective means for the identification of unknown specimens within this species complex. PMID- 8731369 TI - Chronic experimental infection by Trypanosoma cruzi in Cebus apella monkeys. AB - Twenty young male Cebus apella monkeys were infected with CA1 Trypanosoma cruzi strain and reinfected with CA1 or Tulahuen T. cruzi strains, with different doses and parasite source. Subpatent parasitemia was usually demonstrated in acute and chronic phases. Patent parasitemia was evident in one monkey in the acute phase and in four of them in the chronic phase after re-inoculations with high doses of CA1 strain. Serological conversion was observed in all monkeys; titers were low, regardless of the methods used to investigate anti-T. cruzi specific antibodies. Higher titers were induced only when re-inoculations were performed with the virulent Tulahuen strain or high doses of CA1 strain. Clinical, electrocardiographic and ajmaline test evaluations did not reveal changes between infected and control monkeys. Histopathologically, cardiac lesions were always characterized by focal or multifocal mononuclear infiltrates and/or isolated fibrosis, as seen during the acute and chronic phases; neither amastigote nests nor active inflammation and fibrogenic processes characteristic of human acute and chronic myocarditis respectively, were observed. These morphological aspects more closely resemble those found in the "indeterminate phase" and contrast with the more diffuse and progressive pattern of the human chagasic chronic myocarditis. All monkeys survived and no mortality was observed. PMID- 8731370 TI - Analysis of tumor necrosis factor-alpha serum level in Brazilian patients with Dengue-2. PMID- 8731371 TI - Rotaviruses as a cause of nosocomial, infantile diarrhoea in northern Brazil: pilot study. AB - Faecal samples were obtained from 190 children, aged 0 to 5 years, admitted to a public hospital in Belem, Para, Brazil. These patients were placed in a pediatric ward with 40 beds distributed in six rooms. Cases were classified into three groups: (a) nosocomial: children who developed gastroenteritis 72 hr or later after admission; (b) community-acquired: patients admitted either with diarrhoea or who had diarrhoea within 72 hr following admission; (c) non-diarrhoeic: those children who had no diarrhoea three days before and three days after collection of formed faecal sample. Specimens were routinely processed for the presence of rotaviruses, bacteria and parasites. Rotaviruses were detected through enzyme linked immunosorbent assay (ELISA) and subsequently serotyped/electrophoretyped. Rotaviruses were the most prevalent enteropathogens among nosocomial cases, accounting for 39% (9/23) of diarrhoeal episodes; on the other hand, rotaviruses occurred in 8.3% (11/133) and 9% (3/34) of community-acquired and non-diarrhoeic categories, respectively. Mixed infections involving rotavirus and Giardia intestinalis and rotavirus plus G. intestinalis and Entamoeba histolytica were detected in frequencies of 8.6 and 4.3%, respectively, in the nosocomial group. The absence of bacterial pathogens in this category, and the unusual low prevalence of these agents in the other two groups may reflect the early and routine administration of antibiotics following admission to this hospital. Rotavirus serotype 2 prevailed over the other types, accounting for 77.8% of isolates from nosocomial diarrhoeal episodes. In addition, at least five different genomic profiles could be observed, of which one displayed an unusual five-segment first RNA cluster. Dehydration was recorded in all cases of hospital acquired, rotavirus-associated diarrhoea, whereas in only 57% of nosocomial cases of other aetiology. It was also noted that nosocomial, rotavirus-associated diarrhoeal episodes occur earlier (7 days), following admission, if compared with those hospital-acquired cases of other aetiology (14 days). PMID- 8731372 TI - The influence of sugars and amino acids on the blood-feeding behaviour, oviposition and longevity of laboratory colony of Lutzomyia longipalpis (Lutz & Neiva, 1912) (Diptera: Psychodidae, Phlebotominae). AB - Schneider's Drosophila medium, a complex amino acid rich medium was tested alone and with seven different sugars for some aspects of the biology of Lutzomyia longipalpis. Statistically significant results were obtained when sucrose was used alone, indicating that among the sugars tested, this is still the most suitable and practical one for the maintenance of L. longipalpis colonies. However, the addition of Schneider's medium to a pool of different sugars, was suggested to be related with the acceptance of the first and second blood meals and to longevity, these being, obviously, quite relevant aspects when transmission experiments are contemplated. PMID- 8731373 TI - Lignanes from the Brazilian Melia azedarach, and their activity in Rhodnius prolixus (Hemiptera, Reduviidae). AB - A study of the phagoinhibitor and anti-moulting activities of the Brazilian Melia azedarach, collected in the State of Rio de Janeiro, Brazil, allowed the isolation of four lignanes identified as pinoresinol (1), bis-epi-pinoresinol (2), the hemicetal (3) and the diacid (4). These substances are devoid of anti moulting activity. PMID- 8731374 TI - Triatoma infestans is more efficient than Panstrongylus megistus in obtaining blood meals on non anaesthetized mice. AB - We compared the influence of bug density in the capacity of Triatoma infestans and Panstrongylus megistus in obtaining blood meal in non anaesthetized mice. The regression analysis for increase in body weight (mg) versus density (no. of bugs/mouse) showed that in experiments with anaesthetized mice (AM), no correlation was observed. In experiments with non anaesthetized mice (NAM) the weight increase was inversely proportional to density. The regression slope for blood meal size on density was less steep for T. infestans than for P. megistus ( 1.9 and -3.0, respectively). The average weight increase of P. megistus nymphs in experiments with AM was higher than for T. infestans nymphs; however, in experiments with NAM such results were inverted. Mortality of P. megistus was significantly higher than that of T. infestans with NAM. However, in experiments with AM very low mortality was observed. Considering the mortality and the slope of regression line on NAM, T. infestans is more efficient than P. megistus in obtaining blood meal in similar densities, possibly because it caused less irritation of the mice. The better exploitation of blood source of T. infestans when compared with P. megistus in similar densities, favours the maintenance of a better nutritional status in higher densities. This could explain epidemiological findings in which T. infestans not only succeeds in establishing larger colonies but also dislodges P. megistus in human dwellings when it is introduced in areas where the latter species prevails. PMID- 8731375 TI - Life history correlates of adult size in the malaria vector Anopheles darlingi. AB - Adult dry weights of laboratory-reared Anopheles darlingi were highly correlated with wing lengths, which were used to estimate size variation in natural populations of this species. Significant differences in mean wing lengths of females trapped at baits were detected among collections in the same week at one site, but not between three sites in Brazil and Bolivia. Relatively higher variability of wing lengths, compared to collections of other Anopheles (Nyssorhynchus), and platykurtic size distributions in large, single-night collections suggested that An. darlingi females caught at baits emerged from heterogenous larval habitats. No relationship was detected between parous state and the body size of wild-caught females. Adult males and females of laboratory reared An. darlingi did not differ in body size. This absence of sexual size dimorphism is rare among mosquitoes and has not been noted previously in the genus Anopheles. PMID- 8731376 TI - Cytotoxicity of prion protein peptide (PrP106-126) differs in mechanism from the cytotoxic activity of the Alzheimer's disease amyloid peptide, A beta 25-35. AB - The abnormal form of the prion protein (PrPSc), a synthetic prion protein peptide fragment (PrP106-126) and fragments of the Alzheimer's protein precursor, APP, have been shown to be cytotoxic in vitro. We have used synchronous, clonal cell models originally developed to study the toxicity of the Alzheimer's disease amyloid peptide, A beta 25-35, to investigate the actions of PrP peptides. We found that the cytotoxicity of the PrP106-126 depends on its state of aggregation and the cellular expression of PrPc, and is independent of a loss of MTT reduction activity in the absence of cell death associated with the cellular effects of A beta 25-35. These factors may play a role in the lesion specificity of different pathological phenotypes of prion-protein related diseases. PMID- 8731377 TI - Association of A beta 40-positive senile plaques with microglial cells in the brains of patients with Alzheimer's disease and in non-demented aged individuals. AB - To gain insight into the role of microglia in the formation of senile plaques (SP), especially in the generation of the two major molecular species of amyloid beta protein (A beta) with different carboxyl (C)-termini, A beta 40 and A beta 42(43), we conducted double immunolabelling studies on tissue sections from the brains of Alzheimer's disease (AD) and non-demented aged individuals using antibodies to the C-termini of A beta and ferritin, a marker for microglia. All SP were A beta 42(43)-positive in AD as well as in non-demented individuals, only a proportion of which were A beta 40-positive. Both in AD and in non-demented individuals, approximately 2/3 of the A beta 40-positive SP were typical SP with amyloid cores, these being almost invariably associated with microglia. A beta 40 positive, uncored SP were also frequently associated with microglia (mean, 74%), whereas only 24% of A beta 40-negative, uncored SP contained microglia. These results suggest that microglia may play a role in the maturation of SP, especially in the generation of A beta 40. PMID- 8731379 TI - Immunoreactive levels of alpha-ketoglutarate dehydrogenase subunits in Friedreich's ataxia and spinocerebellar ataxia type 1. AB - Enzyme activities of a alpha-ketoglutarate dehydrogenase complex (alpha KGDHC) and one of its constituent subunits, dihydrolipoamide dehydrogenase (E3), are reported to be reduced in non-CNS tissues of some patients with Friedreich's ataxia (FA); however, the results are highly conflicting. To determine whether an enzyme abnormality occurs in brain, we measured immunoreactive levels of the three alpha KGDHC subunits, namely, alpha-ketoglutarate dehydrogenase (E1), dihydrolipoamide succinyltransferase (E2) and E3 in postmortem frontal, occipital and cerebellar cortices of 18 control subjects, 9 patients with FA and, for comparison, 12 patients with spinocerebellar ataxia type 1 (SCA1). Decreased (-20 to -31%) levels of E3 were observed in all three examined areas of the patients with FA with the changes statistically significant in cerebellar and frontal cortices. The E3 reduction could be explained by a loss of alpha KGDHC or other dehydrogenase complexes (e.g. pyruvate dehydrogenase complex) which utilize this subunit. In SCA1, enzyme changes were limited to E2 in cerebellar (-26%) and frontal (-19%) cortices. Although the E3 and E2 reductions are only slight, and may represent secondary events, the changes in this key Krebs cycle enzyme could exacerbate degenerative processes in both of the spinocerebellar ataxia disorders. PMID- 8731378 TI - Acetylcholine muscarinic M2 receptor stimulated [35S]GTP gamma S binding shows regional selective changes in Alzheimer's disease postmortem brain. AB - Oxotremorine-M stimulated [35S]GTP gamma S binding was used to assess acetylcholine muscarinic M2 receptor mediated G-protein function in superior frontal cortical, superior temporal cortical and hippocampal membranes from a series of Alzheimer's disease and matched control subjects. No significant differences were seen in basal [35S]GTP gamma S binding between the groups. The maximal level of oxotremorine-M stimulated [35S]GTP gamma S binding over basal was significantly increased in Alzheimer's disease superior temporal cortex, suggesting an enhanced muscarinic M2 receptor-G-protein coupling efficiency in this region. In contrast, the maximal level of oxotremorine-M stimulated [35S]GTP gamma S binding over basal was unaltered in Alzheimer's disease superior frontal cortex and significantly reduced in Alzheimer's disease hippocampus. Western immunoblotting using antisera towards the alpha-subunits of those G-protein types known to couple muscarinic receptors, revealed that Gq alpha and Gi alpha, but not G(o) alpha, levels were significantly reduced in Alzheimer's disease superior temporal cortex. Neither Gq alpha, Gi alpha, nor G(o) alpha levels were significantly altered in Alzheimer's disease superior frontal cortex or hippocampus. These results suggest that the efficacy of muscarinic M2 receptor G protein coupling shows regional selective changes in Alzheimer's disease postmortem brain with deficits occurring only in a region that shows severe pathology. PMID- 8731380 TI - A comparison of beta-amyloid deposition in the medial temporal lobe in sporadic Alzheimer's disease, Down's syndrome and normal elderly brains. AB - The density of beta-amyloid (A beta) deposits was studied in the medial temporal lobe in non-demented individuals and in sporadic Alzheimer's disease (SAD) and Down's syndrome (DS). No A beta deposits were recorded in six of the non-demented cases, while in a further eight cases, these were confined to either the lateral occipitotemporal or parahippocampal gyrus. The mean density of A beta deposits in the cortex was greater in SAD and DS than in non-demented cases but with overlap between patient groups. The mean density of A beta deposits was greater in DS than SAD consistent with a gene dosage effect. The ratio of primitive to diffuse A beta deposits was greater in DS and in non-demented cases than in SAD and the ratio of classic to diffuse deposits was lowest in DS. In all groups, A beta deposits occurred in clusters which were often regularly distributed. In the cortex, the dimension of the A beta clusters was greater in SAD than in the non demented cases and DS. The data suggest that the development of A beta pathology in the hippocampus could be a factor in the development of DS and SAD. Furthermore, the high density of A beta deposits, and in particular the high proportion of primitive type deposits, may be important in DS while the development of large clusters of A beta deposits may be a factor in SAD. PMID- 8731381 TI - Apolipoprotein E genotype and Alzheimer's disease in an elderly Norwegian cohort. AB - Apolipoprotein E (Apo E) genotyping was performed on an autopsy cohort of neuropathologically verified non-demented controls and subjects with Alzheimer's disease (AD) resident in nursing homes in the Oslo area. AD was associated with a significantly increased frequency of the Apo E epsilon 4 allele; the frequency of the epsilon 2 and epsilon 3 alleles was lower in AD but not significantly so. Age at death in the control group and the AD group did not differ significantly; neither did age at death nor age at onset of dementia in AD vary according to Apo E genotype, though tendencies towards an earlier age at death was seen in individuals with epsilon 4/4 and earlier age at onset dementia in the presence of an epsilon 4 allele and a later age of onset the presence of an epsilon 3 allele were seen. Possession of an epsilon 2 allele had no effect on age at onset of dementia or age at death. Among the possible genotypes there was a trend towards a progression of earliest onset epsilon 4/4, epsilon 2/4, epsilon 3/4, epsilon 3/3, epsilon 2/3 latest onset of dementia and longest duration epsilon 2/4, epsilon 4/4, epsilon 3/4, epsilon 3/3, epsilon 2/3 to shortest duration of dementia. PMID- 8731382 TI - Substantia nigra pars reticulata neurons in Parkinson's disease. AB - We have examined by immunohistochemistry the parvalbumin-containing neurons of the substantia nigra in patients with Parkinson's disease and in age-matched controls. Parvalbumin, a calcium binding protein, is involved in buffering intracellular calcium and in this study was localized within the majority of non pigmented neurons of the human pars reticulata. Previous studies have shown that the parvalbumin-immunoreactive pars reticulata neurons are GABAergic and project to the motor thalamus and tectum. Their increased output, due to the loss of dopaminergic inhibition in Parkinson's disease, decreases cortical activation via thalamic pathways, causing parkinsonian symptoms. In Parkinson's disease there was a significant loss of parvalbumin-immunoreactivity from these neurons, though there was no evidence of actual cell loss. This loss of parvalbumin immunoreactivity was detected only in those cases with end-stage Parkinson's disease. PMID- 8731383 TI - Blood superoxide dismutase, catalase and glutathione peroxidase activities in familial and sporadic amyotrophic lateral sclerosis. AB - Recent studies have implicated free radicals in the pathogenesis of amyotrophic lateral sclerosis (ALS), a fatal, paralytic disorder of motor neurons. Herein we report on measurements of erythrocyte activity of the three main free radical scavenging enzymes: copper/zinc superoxide dismutase (Cu/Zn-SOD), catalase, and glutathione peroxidase. We studied 31 patients with sporadic ALS, 18 with familial ALS, and 24 controls, Mean Cu/Zn-SOD activity was reduced in eight familial ALS patients with mutations of Cu/Zn-SOD but was normal in patients with both familial ALS without identified Cu/Zn-SOD mutations and sporadic ALS. Glutathione peroxidase activity was significantly reduced only in sporadic ALS patients treated with insulin-like growth factor I (100 micrograms/kg). Catalase activity was normal in sporadic and familial ALS. Neither glutathione peroxidase nor catalase activities correlated significantly with duration of symptoms or age at onset. Vitamin E, vitamin C, and beta-carotene did not affect any of the three enzyme activities. These observations indicate that disturbances of catalase and glutathione peroxidase function are not likely to be central factors in the pathogenesis of ALS. PMID- 8731384 TI - The effects of an RNA synthesis inhibitor on the survival and regeneration of rat motoneurones injured at birth. AB - This preliminary study aimed to test the proposal that neuronal death is triggered by expression of specific genes. In rat pups, the sciatic nerve was injured unilaterally on the first day after birth and actinomycin D, an RNA synthesis inhibitor, was administered 3 days later in a lower and higher dose to rat pups just prior to onset of motoneurone death induced by the lesion. Four weeks later, sciatic motoneurones from operated and contralateral pools were counted and their size measured. Significantly fewer motoneurones (16.7% +/- 2.9 SD) survived when the animals were treated with a lower dose of the inhibitor compared to saline treated controls (36.6% +/- 12.7 SD). Experiments recording tension generated in soleus muscle in response to sciatic nerve stimulation, at different ages following nerve crush, suggested that the treatment with the RNA synthesis inhibitor may have delayed regeneration of motor axons back to the muscle. However, survival of motoneurones after treatment with the higher dose did not differ significantly from controls (27.5% +/- 1.3 SD). Nevertheless, the higher dose significantly reduced growth of motoneurones after 4 weeks. Therefore, the higher dose, although impeding normal development of motoneurones, is less neurotoxic than a lower dose. This suggests that a balancing of conflicting effects may have occurred. The neurodegenerative effects of delayed reinnervation induced by RNA synthesis inhibition may be balanced by some neuroprotective effects at a higher dose. More extensive studies are required to validate these pilot findings. PMID- 8731385 TI - Macrophage and microglial cell response after common peroneal nerve cut and crush in C57BL/6J mice. AB - The present study using Mac-1 immunohistochemistry for the detection of macrophages and microglial cells, has investigated the signals for macrophage recruitment in the peripheral nerve fibres and dorsal root ganglia, and microglial cell activation in the dorsal and ventral horns of the spinal cord, at different periods after a right common peroneal (CP) nerve cut or crush in 86 C57BL/6J mice. Though a previous study has demonstrated a delayed regeneration of the peripheral sensory but not the motor fibres in this strain of mice, the present study could not demonstrate a corresponding delay in macrophage recruitment in the L4-L6 dorsal root ganglia and microglial cell activation in the dorsal and ventral horns of the corresponding segments of the spinal cord. In fact, macrophage recruitment and microglial cell activation appeared a short time after the nerve lesion and peaked at 5 days post-operation then subsequently declined. Microglial cells, however, became reactivated at 20-30 days after CP nerve cut, perhaps because of the presence of newly degenerated fibres. In contrast to the above observation, there was no exuberant macrophage recruitment or microglial cell reaction during the period when the majority of the regenerated fibres were detected in the distal segment of the crushed nerve. PMID- 8731386 TI - The effect of oedema and tissue swelling on the measurement of neuroprotection; a study using chlormethiazole and permanent middle cerebral artery occlusion in rats. AB - The effect of chlormethiazole on hemispheric swelling and cortical tissue water content has been investigated in a model of permanent middle cerebral artery (MCA) occlusion. Chlormethiazole (1 mmol/kg i.p.) or saline was administered 60 min after the induction of ischaemia and the animals sacrificed after 24 hours. The cross sectional area of the left hemisphere was increased by 21.8 + 1.9% in saline treated rats, but only by 8.4 + 2.4% in chlormethiazole treated rats. However, the reduction in the absolute area of neurodegenerative damage (mm2) following chlormethiazole administration was considerably greater than the reduction in hemispheric swelling. Cortical tissue water content of ischaemic brain increased from 76.4% to 84.2% and this was attenuated to 78.8% by chlormethiazole administration. These data demonstrate that, providing damage is measured by fitting tissue slices onto prematched stereotactic maps, the decrease in oedema which accompanies a decrease in neurodegeneration does not result in erroneous estimates of neuroprotection. PMID- 8731387 TI - A quantitative and qualitative analysis of prion protein immunohistochemical staining in Creutzfeldt-Jakob disease using four anti prion protein antibodies. AB - Creutzfeldt-Jakob disease (CJD) is the most common spongiform encephalopathy affecting humans. Prion protein (PrP) immunohistochemistry may be useful for studying the localization of prion protein and assessing its role in CJD, the accumulation of a specific protease resistant PrP isoform being apparently pathognomic to the spongiform encephalopathies. However, a number of factors influence the results of immunostaining, making interpretation and comparisons between the staining of different PrP antisera difficult. This study has examined qualitatively and quantitatively the staining produced by four antisera raised to a variety of prion protein homologues in two cases of CJD and two age-matched controls. Quantitative analysis was provided through the use of custom designed image analysis software. Kuru, granular and multicentric plaques, cellular, perivacuolar and white matter PrP deposits were observed in CJD cases with all four antisera. No significant immunostaining was seen in the control tissue. Some antibody specific staining patterns were observed qualitatively; however, quantitative analysis showed statistically significant correlations between all the antisera on the diseased brain tissue. Prion protein immunohistochemistry is thus useful in interpreting patterns of protein distribution in diseased brain but care may be required in interpreting the results of a single antibody. PMID- 8731388 TI - Microwave treatment enhances the immunostaining of amyloid deposits in both the transmissible and non-transmissible brain amyloidoses. AB - The immunolocalization of amyloid deposits containing either protease-resistant prion protein (PrP) or amyloid beta protein (A beta) in the brains of patients with transmissible or non-transmissible cerebral amyloidoses has been greatly facilitated by the pretreatment of tissue sections with concentrated formic acid. We have investigated whether microwave processing of formalin-fixed tissue sections would obviate the need for formic acid pretreatment. Exposure of brain sections to microwaves, even for periods as brief as 1 sec, greatly enhanced the immunostaining of PrP and A beta amyloid deposits in both the transmissible and non-transmissible brain amyloidoses. Microwaving for 1 min yielded staining intensities similar to that following pretreatment with concentrated formic acid for 10 min. Moreover, the combination of formic acid pretreatment and microwave processing resulted in an even more intense staining of amyloid deposits. Microwave processing, which is easy to perform and comparatively inexpensive, makes exposure to the potentially toxic fumes of formic acid unnecessary. PMID- 8731389 TI - Ultrastructural immuno-localization of synthetic prion protein peptide antibodies in 87V murine scrapie. AB - Disease specific forms of a host encoded cell surface sialoglycoprotein called prion protein (PrP) accumulate during this incubation period of the transmissible spongiform encephalopathies. A 33-35 kDa disease specific form of PrP is partially resistant to protease digestion whereas the normal form of PrP can be completely digested. Proteinase K digestion of the murine disease specific form of PrP produces diverse forms of low molecular weight PrP, some of which are N terminally truncated at amino acid residue 49 or 57 within the octapeptide repeat segment. Amyloid plaques are a pathological feature of many of the transmissible spongiform encephalopathies and are composed of PrP. Using synthetic peptide antibodies to the N-terminus of PrP (which is not present in truncated disease specific PrP) and antibodies to the protease resistant fraction of PrP we have immunostained plaques and pre-amyloid deposits in the brains of mice, experimentally infected with the 87V strain of scrapie, for examination by light and electron microscopy. Classical fibrillar amyloid deposits in plaques as well as pre-amyloid deposits were both immunostained by antibodies to the N-terminus of PrP and to the protease resistant core of the PrP molecule. This suggests that both N-terminal and core amino acid residues are present in disease specific PrP released from scrapie infected cells in vivo. The results also suggest that N terminal truncation of PrP may not be essential for formation of amyloid fibrils. PMID- 8731390 TI - The prevalence of Creutzfeldt-Jakob disease. PMID- 8731391 TI - Effects of ipriflavone and its metabolites on human articular chondrocytes cultivated in clusters. AB - Ipriflavone (IP) is an isoflavone derivative that was suggested to have bone sparing effects in post-menopausal and senile osteoporosis. A moderate stimulatory effect of IP and its metabolites on proliferation of osteoblastic cells was reported in rat osteoblastic osteosarcoma cell line. We investigated the effects of different concentrations (0, 1, 10 and 100 micrograms/ml) of IP and its metabolites (MET I, II, III and V) on the incorporation of [3H] thymidine and production of proteoglycans (PG) and type II collagen (COL II) by human articular chondrocytes during a 12-day period, in a three-dimensional chondrocyte culture model. [3H]thymidine uptake was measured in chondrocyte clusters, and specific PG and COL II radioimmunoassays were performed every 4 days on the culture medium and cell clusters. Incubation with IP or its metabolites did not affect [3H]thymidine uptake regardless of the dose. PG released into the culture medium and PG cluster content rose significantly (P < 0.025) in presence of IP (1, 10 and 100 micrograms/ml). MET I increased PG release in culture medium (10 and 100 micrograms/ml) and PG cluster content (100 micrograms/ml). MET II has no effect on PG production. MET III increased PG in culture medium (100 microgram/ml) but did not influence PG cluster content while MET V (100 micrograms/ml) increased both PG release in culture medium and PG cluster content. COL II release in culture medium and COL II cluster content were significantly (P < 0.025) increased in presence of IP (10 and 100 micrograms/ml), MET III (1, 10 and 100 micrograms/ml) or MET V (100 micrograms/ml). MET I and II did not significantly affect COL II production. PMID- 8731392 TI - Experimental osteoarthritis in dogs: a comparison of the Pond-Nuki and medial arthrotomy methods. AB - Lesions induced by transecting the cranial cruciate ligament in two surgical models of osteoarthritis (OA) in mature, male, cross-bred dogs were compared by using an established grading system and alternatives. Previously, we relied on evaluations of lesions in articular cartilage on femurs alone. No statistically significant differences were found between grades for lesions in cartilage when either treated or control joints were compared by surgical method. Because the Pond-Nuki method yielded statistically significant differences between grades for lesions affecting treated and control femurs or tibias, and for some parameters indicative of synovitis, we preferred this method of surgery. Although by using the medial arthrotomy method of surgery, we were able to destabilize the joint in a more consistent manner, significant differences between treated and control joints were found for lesions on tibias, but not femurs, a frequent site for OA in humans. Suggestions are made for enhancing the surgical models and for a more holistic approach to evaluating joints morphologically. PMID- 8731393 TI - Tumor necrosis factor-alpha (TNF-alpha) in canine osteoarthritis: Immunolocalization of TNF-alpha, stromelysin and TNF receptors in canine osteoarthritic cartilage. AB - The presence and distribution of tumor necrosis factor-alpha (TNF-alpha), TNF receptors and stromelysin [matrix metalloproteinase 3 (MMP-3)] in articular cartilage were evaluated in an iatrogenically induced model of osteoarthritis (OA). Eleven adult male dogs were assigned randomly to a control group (N = 4) or an OA group (N = 7). Osteoarthritis was created by surgical transection of the cranial cruciate ligament of one stifle joint. Both femoral condyles were sampled 3 months post-surgery at necropsy and immunohistochemically analyzed for the presence of the aforementioned cytokines and receptors. Chondrocytes stained for TNF-alpha and TNF receptors in control articular cartilage, spanning an area encompassing most of the middle and deep zones. Positive matrical and chondrocytic staining for TNF-alpha, TNF receptors, and stromelysin was present in OA articular cartilage. Staining varied in intensity and distribution and was dependent of the severity of the lesion. Smooth muscle cells of arteries and arterioles (periarticular synovial membrane) were stained for only one (p55) of two TNF receptors; this staining was confined to control tissues. Results indicate that the differential expression of TNF-alpha and its receptors may be important in the normal maintenance of articular cartilage. The increased presence of TNF-alpha and its receptors in articular cartilage with mild osteoarthritic changes suggests a role in the development of early OA. Regulating TNF-alpha may be an important component in the treatment of OA. PMID- 8731394 TI - Parametric and non-parametric measures in the assessment of knee and hip osteoarthritis: interobserver reliability and correlation with radiology. AB - The aim of this study was to evaluate the interobserver reliability of parametric and non-parametric variables in the clinical assessment of hip and knee osteoarthritis (OA). Three rheumatologists examined 49 patients with different radiological stages of OA using different assessment tools such as a tape measure, a goniometer, a plurimeter and a hand-held pull gauge. The reliabilities of parametric variables calculated by analysis of variance (ANOVA) showed much higher values than the non-parametric ones calculated by Kendall's tau beta. The highest levels of correlation in hip OA between clinical functional tests and radiological changes were found for hip extension (r = 0.57; P < 0.01) and the Patrick sign (r = 0.54; P < 0.01) while in knee OA the highest correlations were found for knee circumference (r = 0.5; P < 0.01) and knee flexion (r = 0.035; P < 0.02). Knee muscle strength, as measured with a hand-held pull gauge, showed a high level of interobserver agreement (r = 0.79), but correlated poorly with radiological changes. In conclusion parametric variables of joint morphology as knee circumference of parametric variables of function as the Patrick sign should be preferred for assessing secondary endpoints in OA clinical trials. PMID- 8731395 TI - Interactions of pentosan polysulfate with cartilage matrix proteins and synovial fibroblasts derived from patients with osteoarthritis. AB - Pentosan polysulfate (PPS) has been shown to improve symptoms of patients with osteoarthritis (OA) when studied under double-blinded conditions. Laboratory studies indicated that this drug exhibits multiple actions, including the preservation of articular cartilage (AC) proteoglycans in animal models of OA and the stimulation of hyaluronan synthesis by synovial fibroblasts in vitro and in vivo. As PPS is strongly anionic and has a molecular weight of approximately 5700 Da its ability to enter connective tissues rich in proteoglycans and interact with the resident cells has been questioned. In the present studies, experiments were undertaken to isolate and characterize proteins in human AC which have the potential to bind PPS. Thrombospondin was identified in 4.0 M GuHCl extracts of human AC as a PPS-binding protein. Furthermore, synovial fibroblasts derived from OA joints were shown to secrete thrombospondin and also bind PPS. Using bovine erythrocytes conjugated with PPS a rosetting of the synovial fibroblast could be demonstrated. The level of rosetting was not affected by pre-incubating cultures with thrombospondin antibody suggesting that PPS was interacting directly with the cells. Kinetic studies of 3H-PPS uptake by synovial fibroblasts showed saturation of binding sites within 30 min when cells were maintained at 4 degrees C but preservation of drug uptake for up to 120 min when cells were cultured at 37 degrees C. These data, together with the finding that cells labeled with drug at 37 degrees C showed higher incorporation, than at 4 degrees C after trypsin digestion suggests that PPS first binds to the cell membrane when at 37 degrees C is internalized, possibly by pinocytosis. PMID- 8731396 TI - Effect of methylprednisolone and mechanical loading on canine articular cartilage in explant culture. AB - The objective of this study was to assess the effect of mechanical load on articular cartilage after in vitro corticosteroid exposure. Canine humeral cartilage was equilibrated for 4 days in defined medium with a serum substitute, then exposed to methylprednisolone sodium succinate for 20 h at 0, 0.01 or 1.0 mg/ml. After a drug-free recovery period, the explants were subjected to 0, 1 or 10 mega pascals (MPa) for 1 out every 5 s for 20 min, then incubated with [35S] sulfate and [3H]-leucine for 4 h to measure proteoglycan and protein synthesis, respectively. When the loading occurred 22 h after drug exposure, proteoglycan synthesis was inhibited and protein synthesis, was unaffected by the drug. Both were stimulated by load, relative to controls. When the loading was delayed until 142 h after drug exposure, there was no biosynthetic response to load whether or not the explant had been exposed to the drug. Proteoglycan and protein synthesis 142 h after 0 or 0.01 mg/ml were unchanged or slightly higher than at 22 h, in explants which did not receive load. In contrast, biosynthesis were strongly inhibited 142 h after 1.0 mg/ml, and there was a 40% loss of proteoglycan content, relative to 22 h controls. If explants receiving 1.0 mg/ml also received heavy (10 MPa) loads 142 h later, there was a 17% reduction in total dry content suggesting severe matrix damage. These in vitro results suggest that articular load can help maintain normal cartilage metabolism after corticosteroid exposure, but also suggest that heavy loading after a sub-clinical dose can cause a marked loss of matrix solids. PMID- 8731397 TI - Pulsed electromagnetic fields influence hyaline cartilage extracellular matrix composition without affecting molecular structure. AB - Pulsed electromagnetic fields (PEMF) influence the extracellular matrix metabolism of a diverse range of skeletal tissues. This study focuses upon the effect of PEMF on the composition and molecular structure of cartilage proteoglycans. Sixteen-day-old embryonic chick sterna were explanted to culture and exposed to a PEMF for 3 h/day for 48 h. PEMF treatment did not affect the DNA content of explants but stimulated elevation of glycosaminoglycan content in the explant and conserved the tissue's histological integrity. The glycosaminoglycans in sterna exposed to PEMF were indistinguishable from those in controls in their composition of chondroitin sulfate resulting from chondroitinase ABC digestion. Specific examination with [35S]-sulfate labels showed that PEMF treatment significantly suppressed both the degradation of pre-existing glycosaminoglycans biosynthetically labeled in ovo and the synthesis of new [35S]-sulfated glycosaminoglycans. The average size and aggregating ability of pre-existing and newly synthesized [35S]-sulfated proteoglycans extracted with 4 M guanidinium chloride from PEMF-treated cartilage explants were identical to controls. The chain length and degree of sulfation of [35S]-sulfated glycosaminoglycans also were identical in control and PEMF-treated cultures. PEMF treatment also reduced the amount of both unlabeled glycosaminoglycans and labeled pre-existing and newly synthesized [35S]-sulfated glycosaminoglycans recovered from the nutrient media. [35S]-Sulfated proteoglycans released to the media of both control and PEMF-treated cultures were mostly degradation products although their glycosaminoglycan chain size was unchanged. These results demonstrate that exposure of embryonic chick cartilage explants to PEMF for 3 h/day maintains a balanced proteoglycan composition by down-regulating its turnover without affecting either molecular structure or function. PMID- 8731398 TI - The increased synthesis of inducible nitric oxide inhibits IL-1ra synthesis by human articular chondrocytes: possible role in osteoarthritic cartilage degradation. AB - The degradation of osteoarthritic (OA) cartilage is likely related to the synthesis and the release of catabolic factors by chondrocytes. Nitric oxide (NO) has recently been suggested as playing a role in cartilage degradation. Since NO production is largely dependent on stimulation by IL-1, its effects on factors regulating the IL-1 biological activity, such as IL-1ra, are of the utmost importance. This study examined and compared the level of NO production by normal and OA cartilage and chondrocytes, as well as studied the effect of IL-1-induced NO production on the synthesis and steady-state mRNA of interleukin-1 receptor antagonist (IL-1ra). The NO baseline production by normal cartilage explants was undetectable but inducible by rhIL-1 beta. OA cartilage spontaneously produced NO. About a two-fold increase in NO production was found in OA rhIL-1 beta stimulated (0.5-100 units/ml) cartilage as compared with the similarly stimulated normal cartilage. on chondrocytes rhIL-1 beta-stimulation (0.5-100 units/ml) produced a dose-dependent enhancement of both NO production and IL-1ra synthesis. Treatment with 200 microM N(g)-monomethyl-L-arginine (L-NMA), a well known NO synthase inhibitor, induced over 70% inhibition of the NO production and a marked increased IL-1ra synthesis (average of 84%) and expression (mRNA level). Inhibition of prostaglandin synthesis by indomethacin had no effect on both the NO production or the IL-1ra level. In the present study, we demonstrated the capacity of OA cartilage to produce a larger amount of NO than the normal controls, both in spontaneous and IL-1-stimulated conditions. These data support the notion that, in vivo, OA chondrocytes are stimulated by factors, possibly IL 1, which in turn may induce the expression of NO synthase, thus the synthesis of NO itself. Importantly, our results showed that the elevation of of NO production may be an important factor in the pathophysiology of OA since it can reduce IL 1ra synthesis by chondrocytes. As such, an increased level of IL-1, associated with a decreased IL-1ra level, may be responsible for the stimulation of OA chondrocytes by this cytokine, leading to an enhancement of cartilage matrix degradation. PMID- 8731399 TI - Reproduction of radiographs in scientific articles. Enhancement is not a dirty word. PMID- 8731400 TI - What is your diagnosis? PMID- 8731401 TI - Recent advances in understanding the spectrum of canine generalised progressive retinal atrophy. AB - Canine generalised progressive retinal atrophy (gPRA) is a large and ever increasing collection of naturally occurring, heterogeneous, progressive disorders. Most are inherited in an autosomal recessive manner and new, breed specific forms continue to be described. The gPRAs cause photoreceptor cell death and subsequent retinal degeneration, culminating in blindness. In humans, similar inherited retinal dystrophies are recognised as retinitis pigmentosa and macular dystrophy. Molecular biological studies have revealed disease-causing mutations in several genes in humans and also in mice with retinal dystrophies. Recently, molecular genetic techniques have identified the cause of one form of gPRA in Irish setters while important candidate genes have been investigated in other breeds. Identification of mutations responsible for different forms of gPRA allows carrier and predegenerate animals to be detected using DNA-based tests. Such genetic tests will greatly facilitate the eradication of these diseases in different breeds. PMID- 8731402 TI - Treatment of urinary incontinence in bitches by endoscopic injection of glutaraldehyde cross-linked collagen. AB - Thirty-two spayed bitches with urinary incontinence due to urethral sphincter incompetence, non-responsive to phenylpropanolamine administration, were treated by urethral submucosal injection of glutaraldehyde cross-linked collagen. Urinary incontinence resolved after a single injection in 19 of the bitches. Additional medication with phenylpropanolamine was necessary in five of these dogs, however. Of the 13 bitches that remained incontinent, the injections were repeated in nine. This resulted in a return to continence in five dogs, although two of these required additional medication for complete continence. The cure rate due to collagen injections alone is 53 per cent (17 of the 32 cases). A total of 41 injections were performed and no post-operative complications were observed. PMID- 8731403 TI - Cardiovascular changes associated with anaesthesia induced by medetomidine combined with ketamine in cats. AB - Fifteen cats had anaesthesia induced by intramuscular injection of medetomidine combined with ketamine. By five minutes after drug administration, heart rate had decreased by 31 per cent, respiratory rate had decreased by 70 per cent and systolic blood pressure had increased by 69 per cent. Atipamezole administration was associated with a decrease in systolic blood pressure and an increase in heart and respiratory rates. Time to first head lift was eight minutes and to sternal recumbency 12 minutes after atipamezole administration. Postoperative analgesia was provided by methadone, administered when the cats adopted sternal recumbency. PMID- 8731404 TI - Primary seborrhoea in English springer spaniels: a retrospective study of 14 cases. AB - Primary seborrhoea was diagnosed in 14 English springer spaniels over a 17-year period. Seven of the dogs developed clinical signs by two years of age. The dermatosis began as a generalised non-pruritic dry scaling which gradually worsened. Some dogs remained in this dry (seborrhoea sicca) stage, but in most cases the dermatosis became greasy and inflamed (seborrhoea oleosa and seborrhoeic dermatitis). Eight of the dogs suffered from recurrent episodes of superficial or deep bacterial pyoderma. Histological findings in skin biopsy specimens included marked orthokeratotic hyperkeratosis of surface and infundibular epithelium, papillomatosis, parakeratotic capping of the papillae, and superficial perivascular dermatitis in which lymphocytes and mast cells were prominent. The dogs with seborrhoea sicca responded more satisfactorily to therapy with topical emollient-humectant agents or oral omega-3/omega-6 fatty acid supplementation. Dogs with seborrhoea oleosa and seborrhoeic dermatitis did not respond satisfactorily to topical therapy. One dog, however, responded well to etretinate and omega-3/omega-6 fatty acid administration. No dog was cured. PMID- 8731405 TI - Megaoesophagus secondary to acquired myasthenia gravis. AB - Fifteen dogs with confirmed adult onset idiopathic megaoesophagus, in which no generalised muscle weakness was observed, were tested for the presence of acetylcholine receptor antibodies. Of these, six were found to have values greater than 0.6 nmol/litre, previously determined to be diagnostic of acquired myasthenia gravis. The mean serum titre value for these dogs was 5.59 nmol/litre (range 0.78 to 8.72 nmol/litre). It appears that a significant proportion of dogs presenting with megaoesophagus have myasthenia gravis and, if a prompt diagnosis and appropriate treatment can be instituted, clinical signs may improve. PMID- 8731406 TI - Spontaneous resolution of a possible primary bone tumour. PMID- 8731407 TI - Removal of foreign bodies in the nasopharynx. PMID- 8731408 TI - Intramedullary cisplatin chemotherapy: experience in four dogs with osteosarcoma. AB - Single-agent intramedullary cisplatin chemotherapy provided effective localised control in two of four dogs with advanced stage osteosarcoma unable to withstand an amputation or limb-sparing surgery. Complete remission of the local neoplasm was observed in one of the four dogs, partial remission of the local neoplasm in one dog and progressive disease in the other two. Limb function was preserved in one dog for seven months and that dog was found to be tumour-free when euthanased due to unrelated causes. These preliminary results warrant further investigation into the use of intramedullary cisplatin chemotherapy in the localised management of canine appendicular osteosarcoma. It is possible that this mode of treatment may be combined with other treatments to maximise the survival (i.e., systemic control) in other dogs affected with appendicular osteosarcoma. PMID- 8731409 TI - Pericardial effusion in a dog with rhabdomyosarcoma in the right ventricular wall. AB - A seven-year-old male labrador retriever presented in right heart failure with weak femoral pulses, and pleural, abdominal and mild pericardial effusion. No diagnosis could be established initially. Two days later, the dog developed severe pericardial effusion causing cardiac tamponade. A tumour in the right ventricular wall was visualised on ultrasonographic examination. An exploratory thoracotomy was performed and biopsies of the mass submitted for histopathological examination. A diagnosis of rhabdomyosarcoma arising from the myocardium was established. Cardiac rhabdomyosarcoma has been reported in only two dogs. Neither report was associated with pericardial effusion. PMID- 8731410 TI - Feline spongiform encephalopathy. BSAVA Scientific Committee. PMID- 8731411 TI - Periprosthetic bone density in uncemented femoral hip implants with proximal hydroxylapatite coating. AB - The short term results of a prospective dual energy x-ray absorptiometry (DEXA) study of periprosthetic bone remodeling around titanium femoral stems proximally coated with hydroxyapatite are presented. Thirty patients underwent DEXA scanning on the 15th and 45th postoperative days, 12th, 18th postoperative month. DEXA showed global femoral periprosthetic remodeling during the first 18 postoperative months. However, no significant variation in bone mineral density occurred around the lateral metaphyseal regions, where cancellous bone is greatest. PMID- 8731412 TI - Modular prostheses in metastatic bone disease of the proximal femur. AB - In the period from 1988 to 1992, 23 patients (16 females and 7 males, average age at operation 57.3 years) with metastatic disease of the proximal femur were treated by resection of the lesion and insertion of a modular prosthesis (PGR). At latest review (at an average 3.2 years following the operation, range 1 to 5 years) twelve patients were still alive. Local recurrence of the neoplasia occurred at, respectively 6, 8, and 12 months after prosthesis insertion in the three patients in whom surgery was performed because of a pathological fracture. Pain relief was obtained in all cases after surgery and no patient developed any complication during or after surgery. Functional results (Enneking Function Evaluation system) were excellent in 2 patients, good in 13 and fair in 8. PGR modular prostheses appear to be a safe form of palliative treatment in a patient with proximal femoral metastases. PMID- 8731413 TI - Radiographic variables in normal and osteoarthritic hips. AB - The width of the weight-bearing surface (WBS) of the acetabulum and its obliquity; height, width, opening angle and alpha angle of the gothic arch; diameter of femoral head; the angle between the center of the femoral head and the most lateral point of the acetabulum (CE angle); neck-shaft angle (CCD angle); offset; angle of aperture, surface area, and volume of the spherical sector; radius and height of the spherical bowl; and distance from teardrop were assessed to study mechanically normal and frankly osteoarthritis hips. There appears to be a direct correlation between primary alterations of the above variables and development of osteoarthritis of the hip. PMID- 8731414 TI - Early ultrasound diagnosis of developmental dysplasia of the hip. AB - The results of ultrasound diagnosis of developmental dysplasia of the hip (DDH) in several studies of 1,000 and 1,332 consecutive patients, performed according to Graf's method from 1984 to the present, are presented. Prior to 1984 the authors used radiography exclusively in suspected cases of DDH, however, the change from radiography to ultrasonography resulted in significant improvement in the ability to diagnose and treat DDH. Different standard methods of treatment have been developed for the management of each morphological type of the newborn's and infant's hip. Ultrasound is more sensitive than clinical assessment for diagnosing DDH since it detects the dysplastic morphology of the hip when Ortolani's test is negative. Instability can be demonstrated directly. The success of treatment of DDH depends on the time of diagnosis the early diagnosis obtained by ultrasound screening can significantly improve the results of treatment. PMID- 8731415 TI - High resolution real time ultrasonography in antenatal diagnosis of skeletal anomalies. AB - Two protocols were used for the ultrasonographic antenatal diagnosis of skeletal anomalies. The first was less selective and was used in pregnancies with a low risk of skeletal malformations. The second was applied to higher risk pregnancies. Both involved several ultrasound examinations, according to the patient's past medical history. In the period 1988 to 1993, 740 pregnant women in whom fetal malformations were suspected underwent serial ultrasound scanning. In 56 (7.5%), a skeletal anomaly was identified. The use of two protocols offers wide applicability and acceptable reliability in screening congenital skeletal problems in the fetus. PMID- 8731416 TI - Fetal chondral homografts in the repair of articular cartilage defects. AB - Lesions of the articular cartilage were bilaterally induced in the patella and the femoral condyle of adult NZW rabbits. A fresh fetal chondral homograft was implanted in one side, the untreated side serving as control. Fetal chondral tissue appeared to survive when implanted at the site of articular injury in adult animals, with no histological evidence of cell-mediated immune response. Fetal chondroblasts showed some signs of maturation and induced endochondral ossification at the base of the graft, with sound anchoring of the graft to the host tissue. In most cases, the homograft fused directly with the adjacent articular cartilage restoring the continuing of the articular surface. However, at implantation the graft could not be easily kept in place, and therefore some grafts had poor mechanical stability. Condylar grafts were incorporated better than patellar grafts. The healing process seemed to be related to the mechanical stability of the graft and to the anatomical site of injury. PMID- 8731417 TI - Achilles and patellar tendinopathies in athletes: pathogenesis and surgical treatment. AB - In the period from 1985 to 1992, 136 athletes underwent 148 surgical exploration of Achilles (98 operations in 90 patients) and patellar (50 operations in 46 patients) tendons for persistent pain. The morphology and the biochemistry of tendon samples taken at operation were investigated. A marked increase of glycosaminoglycans, especially chondroitin sulphate, was present. The complete alleviation of pain and the return to full activity were obtained in 79% of the athletes, while the remaining 21% were not able to return to their sports activities. Generally, successful outcome after surgery was experience by younger patients who had tendon pain of short duration--which was correlated to a lesser degree of tendon degeneration. PMID- 8731418 TI - Percutaneous longitudinal tenotomy in chronic Achilles tendonitis. AB - In the period from August 1989 through December 1992, 42 track and field athletes with unilateral Achilles tendonitis and/or peritendinitis were operated using multiple percutaneous longitudinal tenotomy under local anesthesia following failure of conservative treatment. Patients were reviewed in a special clinic over three consecutive days at an average of 19.2 (9.4) months from the operation. Of the 42 patients operated, 36 (86%) attended. On this occasion, only subjective assessment was carried out. Of the other six patients, four were interviewed by telephone using the same questionnaire; two were lost to follow up. The results in 25 patients (59.5) were rated as excellent, 11 (28.2%) as good, 4 (9.5%) as fair, and 2 (4.8%) as poor. PMID- 8731419 TI - Thumb reconstruction. AB - Experience in 161 thumb reconstructions are reported. The results of pollicizations are evaluated according to function, cosmesis, and two-point discrimination. Of 49 pollicization, 32 were considered excellent, 10 very good, 5 good, and 2 fairly good. In the 16 wrap-around procedures, all showed a very good (less than 6 mm) or good (6 to 12 mm) two point discrimination. Similar results were seen after toe-to-hand transfers when careful nerve repair had been performed. PMID- 8731420 TI - Callotasis lengthening: a review of some technical aspects. AB - Limb lengthening by gradual osteotomy distraction has been practiced since the beginning of the century. The results have not always been uniformly good because of the invasiveness and too sudden elongation applied. Using the callotasis technique, a low energy osteotomy is produced in a metaphyseal area. The periosteum is spared, and distraction, generally at a rate of 0.25 mm every six hours, is started 7 to 14 days after the osteotomy. Using the principle of tension-stress outlined by Ilizarov in the early 1950s, it is possible to produce consistent lengthening of a given bone segment. However, great attention to detail is necessary, as limb lengthening techniques show a steep learning curve, with a significant initial complication rate. PMID- 8731421 TI - Locked nailing of the femur and tibia: minimizing operative difficulties. AB - The technique of closed intramedullary nailing has inherent operative difficulties and complications, which are especially evident and costly to the surgeon just beginning to become familiarized with the procedure. Practical advice about the techniques of closed femoral and tibial nailing to improve awareness of the possible problems is provided. The nailing procedure is analyzed in a step-by-step fashion from pre-operative planning to locking. Accurate pre operative planning for the procedure to run smoothly is mandatory. PMID- 8731422 TI - Minimal trauma knee hemarthrosis. AB - Four patients who suffered from minimal trauma knee hemarthrosis are presented. In all patients, the cause of the hemarthrosis was not the traumatic event in itself, but an underlying clotting or collagen disorder. The necessity of interaction between the orthopaedic surgeon and a coagulation unit is stressed. PMID- 8731423 TI - Arterial embolization of aneurysmal bone cysts in children. AB - The long-term results of six aneurysmal bone cysts in children treated by selective arterial embolization are reported. All the cysts were large and difficult to treat by surgery. To date, no surgical treatment has been required in these children and an almost complete restoration of the original shape of the affected bones has been achieved. PMID- 8731424 TI - Increased density of dopamine D5 receptor in peripheral blood lymphocytes of migraineurs: a marker for migraine? AB - The expression of dopamine D5 receptor was investigated in peripheral blood lymphocytes of 11 migraine patients and of ten healthy control subjects using a radioligand binding technique with [3H]SCH 23390 as a ligand. [3H]SCH 23390 is a benzazepine derivative with potent antagonist properties at the dopamine D1-like receptors. [3H]SCH 23390 was specifically bound to peripheral blood lymphocytes of migraineurs and control subjects in a manner consistent with the labelling of a dopamine D5 receptor. In migraineurs a statistically significant higher density of lymphocyte dopamine D5 receptor compared with controls was noticeable, whereas the affinity of the radioligand was unchanged. The increased density of dopamine D5 receptor in peripheral blood lymphocytes may reflect the dopaminergic hypersensitivity displayed by migraineurs and may represent a relatively simple and reliable peripheral marker of altered dopaminergic function. PMID- 8731426 TI - The carbohydrate N-acetylglucosamine is involved in the guidance of neurites from chick ciliary ganglion neurons through the extracellular matrix of rat skeletal muscle fiber. AB - In the present work we study the functional role of muscle cell extracellular matrix components in axonal guidance during synaptic regeneration. We focused on components recognized by the N-acetylglucosamine-specific lectin called wheat germ agglutinin (WGA) that has been shown to bind to the muscle cell extracellular matrix. We have used a cryoculture bioassay which is based on the ability of chick ciliary ganglion neurons to grow on rat skeletal muscle cryostat sections [Covault, J., et al., J. Neurosci., 105 (1987) 2479-2488.]. In control cultures neurites extended upon the muscle sections closely associated to the muscle cell surface. Masking WGA lectin receptors on the muscle cell surface perturbed the behavior of neurites. On WGA-treated sections, most of the neurites extended indiscriminately on intercellular and intracellular regions. These results indicate that N-acetylglucosamine-bearing molecules on muscle cell surfaces may play functional roles in the guidance of neurites through the extracellular matrix. PMID- 8731425 TI - High affinity block by nimodipine of the internal calcium elevation in chronically depolarized rat cerebellar granule neurons. AB - Chronic depolarization enhances survival of cultured rat cerebellar granule neurons by elevating the internal calcium concentration ([Ca2+]i). In Fura2 loaded cells maintained in 25 mM KCl, [Ca2+]i was close to 150 nM and decreased to approximately 50 nM when KCl was lowered to 5.4 mM. The effect of nimodipine (IC50 = 0.45 nM) was similar to depolarization removal, while agatoxin IVA (up to 500 nM) was ineffective. In whole-cell-clamp experiments, the IC50 for current inhibition was 57 nM, while with transient, KCl-induced depolarizations, the dose dependence of nimodipine inhibition had a 'double-affinity' shape, with IC50(1) = 0.30 nM and IC50(2) = 71 nM. We concluded that L-type calcium channels are the main responsible of the elevated internal calcium level necessary for survival in these neurons. These channels do not inactivate and bind nimodipine with different affinity, depending on their state. PMID- 8731427 TI - Innervation of rat brainstem motoneurones in organotypic culture from a co cultured sensory explant. AB - A co-culture procedure was used to investigate the formation of target-specific afferents on brainstem motoneurones of the rat in organotypic culture. Explants of brainstem motor nuclei together with a tongue explant were co-cultivated with explants obtained from vestibular, collicular or sensory trigeminal nuclei. Fiber connections between sensory and motor explants were observed after a few days in culture. Fluorescent labelling with carbocyanin dyes showed that fibers from the sensory explants invaded the motor explant, whereas no axon or dendrites from the motor explant were found penetrating the sensory structures. A double fluorescent labelling procedure, involving retrograde labelling of motoneurones and orthograde labelling of sensory fibers, demonstrated that sensory fibers come in close contact with labelled motoneurones. Electrical stimulation of the sensory explant induced muscle contraction, thus demonstrating that afferent fibers from the co-explanted structure establish functional synaptic connections with motoneurones. PMID- 8731428 TI - Preventive effect of melatonin against cyanide-induced seizures and lipid peroxidation in mice. AB - Subcutaneous injection of potassium cyanide (6, 8, and 9 mg/kg) caused a severe tonic seizure in a dose-dependent manner. However, the incidence of seizures induced by potassium cyanide was significantly inhibited by preadministration of melatonin (20 mg/kg, s.c.) Lipid peroxidation in homogenates from whole brain of mice was significantly increased (39%, 106% and 132%, respectively) by the exposure to potassium cyanide (0.01, 0.05, and 0.1 mM). The potassium cyanide (0.1 mM)-induced lipid peroxidation was prevented by melatonin (0.1, 0.5, 1.0, or 2 mM) in a concentration-dependent manner. These results suggest that free radicals formation and subsequent lipid peroxidation may contribute in part to the development of seizures induced by cyanide in mice. PMID- 8731429 TI - Influence of mediobasal hypothalamic lesion and catecholamine receptor antagonists on ultradian rhythm of EEG in the posterior hypothalamus of the rat. AB - The delta and theta frequency bands of the EEG in the posterior hypothalamic area (PH) of the urethane-anaesthetized rat vary according to an ultradian rhythm with a frequency of approximately one cycle per 100 min. Injected into the lateral ventricle, prazosin (150 nmol) abolished the rhythmic changes, propranolol (150 nmol) increased, while yohimbine, SKF-83566 and sulpiride (150 nmol each) decreased the cycle duration. Electrocoagulation of the rostral arcuate nucleus and median eminence (Arc-ME) of medial basal hypothalamus abolished the rhythmic EEG changes in the PH. Our results indicate that the ultradian EEG rhythm in the PH is susceptible to regulatory influences mediated by noradrenergic and dopaminergic neurons. For the generation of the ultradian rhythm, the functional integrity of the Arc-ME is required. PMID- 8731430 TI - Hyperpolarizing shift of the M-current activation curve after washout of muscarine in bullfrog sympathetic neurons. AB - The mechanism underlying the over-recovery of an M-type potassium current following the washout of muscarine (20 microM) has been examined. Whole-cell recordings were made from single neurons dissociated from bullfrog sympathetic ganglia. During over-recovery, the maximum M-conductance decreased by about 2.8 nS while the steady-state M-current activation curve was displaced in the hyperpolarizing direction by about 13 mV. These data suggest that a hyperpolarizing shift in the kinetics of M-current causes over-recovery in amphibian autonomic neurons. PMID- 8731431 TI - Activation of GABAergic function necessary for afterdischarge generation in rat hippocampal slices. AB - Involvement of gamma-aminobutyric acid (GABA)-mediated function in epileptogenesis was studied in rat hippocampal slices, in which repetition of high-frequency electrical stimulation induced afterdischarges (ADs) to create an in vitro model for ictal activity. A GABAA receptor antagonist, bicuculline, fully blocked the ADs. On the other hand, the presence of bicuculline caused single stimuli to evoke short-duration epileptiform bursts, a well-known model for interictal activity. Therefore, we conclude that activation of GABAergic function appears to be necessary for ictal activity, while its dysfunction induces interictal activity, and propose a modification to the simple disinhibition hypothesis for epileptogenesis. PMID- 8731432 TI - Central administration of a nitric oxide synthase inhibitor impairs spatial memory in spontaneous hypertensive rats. AB - Nitric oxide is widely recognized as a putative retrograde messenger in the brain. We infused NG-monomethyl-L-arginine (L-NMMA; 25 mg/kg), an inhibitor of nitric oxide synthase (NOS), continuously for a week into the dorsal third ventricle (D3V) of spontaneous hypertensive rate (SHR) by an osmotic infusion pump. Rats administered with L-NMMA showed impaired performance of a radial arm maze task compared with control rats administered with saline. We observed significant reductions of the NOx level in the cerebrospinal fluid (CSF) of the rats administered with L-NMMA, but not in the control rats. Both groups showed no change in systolic blood pressure or serum NOx level. The results provide evidence of a more specific effect of NOS inhibition to the brain independent of alterations in the systemic hemodynamics. PMID- 8731433 TI - Induction of interleukin-1 beta mRNA in the hypothalamus following subcutaneous injections of formalin into the rat hind paws. AB - The induction of interleukin-1 beta (IL-1 beta) mRNA in the rat brain following subcutaneous injection of formalin into the hind paws was investigated by in situ hydridization. IL-1 beta mRNA was markedly induced in the hypothalamus after the injection of formalin into both hind paws. On the other hand, IL-1 beta mRNA was scarcely observed in the hypothalamus of saline-injected control rats. The type of cells expressing IL-1 beta mRNA was likely glia because their nuclei were densely stained by Cresyl violet and were relatively small. The present results suggest that IL-1 beta mRNA is induced in the glial cells of the hypothalamus by persistent pain which is caused by formalin injection. PMID- 8731434 TI - Evidence for apoptotic cell death in the choroid plexus following focal cerebral ischemia. AB - Focal cerebral ischemia in rats subjected to middle cerebral artery (MCA) occlusion results in apoptotic DNA fragmentation and activation of putative cell death effector genes in neurons and functional impairment of the plexus choroideus. In the present study we investigated whether cerebral ischemia may induce apoptotic cell death in the choroid plexus. Using in situ end-labeling by terminal transferase and fluorescein-dUTP, nuclear DNA breaks were detected in the choroid plexus of the lateral ventricle of the ischemic hemisphere after 6 h but not after 1.5 h of MCA occlusion. Intense cytoplasmic immunostaining for pro apoptotic Bax protein and moderate immunolabeling for Bcl-X was observed in the epithelium of the choroid plexus of the lateral and third ventricles. However, constitutive expression of Bax and Bcl-X proteins in the plexus choroideus did not change significantly following focal ischemia. Thus, cells of the choroid plexus may die by apoptosis after several hours of cerebral ischemia. Modulation of cell death effector genes of the bcl-2 family however, may not be required for apoptotic cell death to occur. PMID- 8731435 TI - Differential increases in rat retinal ganglion cell size with various methods of optic nerve lesion. AB - Optic nerve injury is a well established paradigm for studying a variety of neuronal responses, although the actual method of nerve severance is rarely taken into account. This study assessed changes to ganglion cell size in three different methods of optic nerve lesions. Adult rats underwent either one of two types of mechanical nerve crush, or an axotomy. Ganglion cells were visualised by retrograde labelling from the optic nerve with 4Di-10ASP, and soma size measured. Two weeks after lesion, mean soma size was increased in all groups. However, at 4 weeks, the crush groups continued to show an increase (60.5% larger than normal), while the mean cell size in the axotomy group was almost at normal levels (0.2% smaller than normal). This study supports the hypothesis that axotomy, and not simple crush, deprives ganglion cells of substances beneficial to cell survival. PMID- 8731436 TI - Nicardipine enantiomers inhibit calcium and outward currents in rat pars intermedia cells. AB - The whole-cell patch clamp technique was used to investigate the effect of the dihydropyridine antagonist nicardipine and its two enantiomers on calcium and outward currents in rat pars intermedia cells. Inward voltage-activated calcium currents as well as outward currents were blocked by nicardipines (enantiomers and racemic substance). We found a highly significant correlation between the blockage of voltage-activated currents and outward currents, indicating an association between recorded currents. Therefore, the inhibitory effect of nicardipines was measured as outward current inhibition. The effects of (-) and (+) enantiomers as well as the racemic nicardipine were concentration-dependent. The (+)-enantiomer was the least potent with an ED50 of 3.8 +/- 1.2 microM, while the (-)-enantiomer inhibited currents with an ED50 of 0.7 +/- 0.1 microM. Racemic nicardipine blocked currents with an ED50 of 2.2 +/- 0.6 microM. PMID- 8731438 TI - Altered mitochondrial membrane fluidity in AD brain. AB - Oxidative damage on biological membranes has been proposed as a cause of the alterations observed in aging brain and, more severely, in Alzheimer's disease (AD). In this study we evaluated membrane fluidity of mitochondria extracted from different areas of normal and AD brains by means of fluorescence polarization technique. AD mitochondria showed a significant reduction of membrane fluidity compared to controls except in cerebellum. This might be caused by a greater lipid peroxidation of biological membranes, as suggested by in vitro experiments we performed to this purpose. From these results the possible role of oxidative stress in AD pathogenesis is supported. PMID- 8731437 TI - Evidence for the presence of NK1 and NK3 receptors on cholinergic neurones in the guinea-pig ileum. AB - In a guinea-pig ileum longitudinal muscle preparation, substance P (SP) (> or = 6 nM) caused an initial contraction followed by a sustained plateau contraction of about 20-50% of the initial response. This plateau contraction is caused by the SP-induced activation of cholinergic motoneurones which contract the smooth muscles by the released acetylcholine (ACh). We investigated the contribution of neurokinin NK1 and NK3 receptors during this 'plateau phase' of contraction. The plateau contraction induced by SP (60 nM) was significantly reduced by the NK1 receptor antagonist CP-96,345 (200 nM) added 5 min after SP, but was not affected by its inactive enantiomer CP-96,344 (200 nM). The NK1 receptor antagonist CP 99,994 (100 nM) significantly reduced the plateau contraction induced by SP (60 nM and 600 nM) and that induced by the NK1 receptor agonist substance P-O methylester (SPOMe; 100 nM). CP-99,994 (100 nM), however did not affect the plateau contraction induced by the NK3 receptor agonist [Asp5,6, MePhe8]-SP(5-11) (100 nM). The plateau contraction induced by SP (600 nM) was not affected by the NK3 receptor antagonist SR-142,801 (100 nM), added 5 min after SP. Pre-incubation of the ileum with SR-142,801 (100 nM) 30 min prior to the addition of SP (600 nM) also had no significant effect on the plateau contraction. However, it significantly reduced the ileal contraction in the first minutes after the initial spasmogenic contraction. We suggest that SP induces the plateau contraction of the guinea-pig ileum longitudinal muscle mainly by the activation of NK1 receptors on cholinergic neurones. PMID- 8731439 TI - Temporal control of the reach and grip components during a prehension task in humans. AB - The kinematic changes of the manipulation and transport components during a prehensile movement were examined in an experimental condition that required alterations during the manipulation phase. Subjects started with a normal finger posture but were required to briefly open and close their thumb and index finger while transporting the wrist to grasp the object. The changes in the aperture altered most of the kinematic landmarks of the wrist. There was considerable slowing of most kinematic characteristics for the wrist transport, and the velocity profile of the wrist showed two peaks. The time of occurrence of the first peak was poorly correlated with the time of the first maximal finger aperture (before the brief finger touch). In contrast, the second wrist velocity peak was highly correlated with the time to the second maximal finger aperture (just before the grasp of the object). Thus, as the reach progressed and the object to be grasped was approached, the temporal relationship between the wrist and aperture components of the movement became more temporally related. PMID- 8731440 TI - Adenoviral-mediated gene transfer into guinea pig cochlear cells in vivo. AB - Loss of ganglion cells is a common and irreversible complication of hair cell loss in the cochlea. Gene transfer could potentially be used to prevent this neuronal degeneration and other pathologies in the cochlea. Human adenoviruses should provide a feasible gene transfer vehicle for transducing the quiescent cochlear neurons and organ of Corti epithelium. We now describe in vivo experiments in which a replication-deficient adenoviral vector, Ad.RSVntlacZ was injected into the perilymphatic fluid of six normal guinea pigs. Postoperative recovery of animals was complete. Inner ear tissues were assessed for histology and for presence of lacZ-positive cells 1 or 2 weeks after the injection. A large number of blue (lacZ-positive) cells were observed in the neural, epithelial and connective tissues of the cochlea. In four ears spiral ganglion cell infection exceeded 50%, throughout the length of the cochlear spiral. No major pathology was detected in the organ of Corti and other cochlear tissues, and no infection was present in the vestibular tissues or the contralateral cochlea. Immunocytochemical assessment of T cells revealed an increased in the number of lymphocytes in the connective tissue lining the perilymphatic spaces. We conclude that efficient gene transfer into multiple types of cochlear cells in vivo can be achieved without major morphological signs of pathology or toxicity. PMID- 8731441 TI - Abdominal lymphadenopathy in benign diseases: sonographic detection and clinical significance. AB - We prospectively evaluated the frequency of lymphadenopathy in the right upper abdominal quadrant as detected by sonography in 650 consecutive unselected patients, after excluding patients with a known lymphoma or abdominal carcinoma and patients with acquired immunodeficiency disease. Evidence of enlarged lymph nodes (few in number, with an elongated shape and isoechoic to the liver, 8 to 22 mm in size), found primarily in the gastrohepatic ligament and porta hepatis, was seen on sonographic scans in 106 patients (16.3%). Associated conditions in 69 of 106 patients (65%) were hepatobiliary or pancreatic diseases and, less frequently, other benign entities (12 patients; 11.3%); in 25 cases (23.5%) no significant abdominal or systemic disease was present. Comparison with CT or surgical findings, or both, was available in 36 cases. We conclude that lymphadenopathy in the right upper abdominal quadrant may be found in relation to different non-neoplastic conditions as well as in the absence of any significant intra-abdominal disease. The frequency of this finding on sonographic scans must be recognized to prevent misdiagnosis of lymphoma or metastatic disease as well as to avoid overstaging of local (hepatobiliary, pancreatic, gastric) neoplasms. PMID- 8731442 TI - Transrectal sonography of the female urethra in incontinence and frequency urgency syndrome. AB - Transrectal sonography of the bladder and urethra was performed to investigate urethral conditions in 296 women (191 with stress urinary incontinence, 78 with frequency-urgency syndrome, and 27 who were asymptomatic). Patients with stress urinary incontinence showed larger pubovesical angle during both resting and straining states than those with either frequency-urgency syndrome or those who were asymptomatic. Pubovesical length did not vary among the three groups of women. Five types of stress urinary incontinence were classified according to the four findings of transrectal sonography: hypermobility of the bladder neck, bladder neck incompetence, urethral sphincteric incompetence, and the presence of a cystocele. Of the patients with frequency-urgency syndrome, 55.1% showed bladder neck incompetence, whereas only 29.6% of the asymptomatic women did. Of the patients with frequency-urgency syndrome, nine had urethral sphincteric incompetence and one had a cystocele. none of the asymptomatic women had any of these disorders. However, the presence of bladder neck incompetence did not correlate well with the presence of clinical symptoms. Transrectal sonography could not differentiate symptoms caused by the bladder from those caused by the urethra. Although a large overlapping of pubovesical angle size was noted among the groups of women, the angle was largest in those with stress urinary incontinence, second largest in those with frequency-urgency syndrome, and smallest in those who were asymptomatic. These findings held true during both resting and straining. Bladder neck incompetence was often associated with a weak extrinsic continence mechanism, as was demonstrated by pubovesical angle changes in both frequency-urgency syndrome patients and asymptomatic women. Urethral sphincteric incompetence was not present in asymptomatic women but could be found in 12.8% of those with frequency-urgency syndrome. PMID- 8731443 TI - Persistent intrahepatic right umbilical vein: a prenatal sonographic series without significant anomalies. AB - Umbilical venous anatomy variation is considered extremely rare. The intrahepatic persistent right umbilical vein represents one version in the spectrum of umbilical vein variations. Prior reports of a prenatally diagnosed persistent right umbilical vein suggest it is strongly associated with severe fetal anomalies including congenital heart disease and gastrointestinal malformations. Our experience contrasts with these findings. Nine cases of intrahepatic persistent right umbilical vein were identified via prenatal sonography between July 1992 and January 1994. The gestational ages of the fetuses ranged from 20 to 36 weeks. Targeted fetal surveys were performed in all nine fetuses and formal fetal echocardiograms were obtained in six. All nine infants have been delivered. In nine cases, the intrahepatic persistent right umbilical vein was an isolated observation with no abnormalities identified pre- or postnatally except one case of hypospadias. Formal fetal echocardiography was normal in all six of the fetuses studied. Given previous reports describing a high risk of congenital malformations in association with the presence of a persistent right umbilical vein, the sonographic finding of this anomaly should still prompt targeted fetal sonography and fetal echocardiography. However, our series indicates that this vascular variant may occur as an isolated abnormality and may not necessarily represent the ominous finding suggested previously. PMID- 8731444 TI - Dolichocephaly and oligohydramnios in preterm premature rupture of the membranes. AB - This study evaluates the association between dolichocephaly and fetal outcome after preterm premature rupture of membranes. Dolichocephaly was more common in preterm fetuses in the breech presentation than those in the cephalic presentation and was more common in fetuses with oligohydramnios of long duration. Of fetuses in the cephalic presentation, 10 of 12 (83%) surviving dolichocephalic fetuses had respiratory distress syndrome compared with 31 of 73 (42%) normocephalic fetuses. However, other outcome parameters did not differ significantly. We conclude that the finding of dolichocephaly is associated with oligohydramnios of long duration. In fetuses with preterm premature rupture of membranes it is associated with respiratory distress syndrome, but not otherwise with a poor neonatal prognosis. PMID- 8731445 TI - Age-independent assessment of abnormal fetal growth in the third trimester. AB - This study assessed the use of the sonographic growth rate of a fetus in the third trimester independently of its gestational age as a tool to evaluate adequacy of fetal growth. Ultrasonographic records were reviewed to calculate our third trimester growth rate, and three rates of fetal growth were established. One hundred and thirty-three patients having two third trimester sonographic examinations were then studied, and our average, above average, and below average growth rate zones were found to be excellent predictors for fetal size at birth. The below-average zone identified fetuses who were not clinically suspected of having IUGR but were determined to be SGA at birth. PMID- 8731446 TI - Severity of polyhydramnios does not affect the prevalence of large-for gestational-age newborn infants. AB - The purpose of this study was to evaluate the relationship between the severity of polyhydramnios with or without maternal diabetes and the prevalence of large for-gestational-age newborn infants. A case control design was used. The study group consisted of 275 singleton pregnancies with an amniotic fluid index > or = 25.0 cm. An equal number of controls was matched for maternal age, gravidity, parity, and gestational age. Polyhydramnios was categorized into three groups by severity: mild (amniotic fluid index 25 to 30 cm.); moderate (amniotic fluid index, 30.1 to 35.0 cm); and severe (amniotic fluid index > or = 35.1 cm). Among our study group, 72.7%, 19.7%, and 7.6% of cases had mild, moderate, and severe polyhydramnios, respectively. Patients with polyhydramnios had a significantly higher prevalence of large-for-gestational-age neonates (27%) than did controls (10%) (P < 0.001). No correlation was seen between the severity of polyhydramnios and neonatal delivery weight. The prevalence of gestational and class > or = B diabetes mellitus was significantly higher among patients with polyhydramnios (17.7%) than among controls (7%) (P < 0.003). Once polyhydramnios was diagnosed sonographically, however, maternal diabetic status did not affect the prevalence of large-for-gestational-age newborn infants. We conclude that the prevalence of large-for-gestational-age neonates is 2.7 times greater when polyhydramnios is present than when the amniotic fluid volume is normal. Neither the severity of polyhydramnios nor the presence of maternal diabetes mellitus strengthens the relationship between polyhydramnios and large-for-gestational-age newborn infants. PMID- 8731447 TI - Isolated fetal choroid plexus cysts and karyotype analysis: is it necessary? AB - The purpose of this study was to evaluate the risk of fetal aneuploidy in the presence of isolated choroid plexus cysts and to evaluate the results obtained from our institution and those reported previously in the English literature. All patients with fetal choroid plexus cysts on prenatal ultrasonography were offered genetic counseling and amniocentesis for fetal karyotyping. Seven of 274 fetuses, 2.6% (95% confidence interval = 1.0 to 5.2%), with isolated choroid plexus cysts were aneuploid. Literature analysis located 23 other reports of 1537 fetuses with isolated choroid plexus cysts; 26 were karyotypically abnormal, 1.7% (95% confidence interval = 1.0 to 2.4%). When evaluating only those patients whose indication for amniocentesis was choroid plexus cysts (i.e., eliminating those patients with advanced maternal age or abnormal serum screening) the risk of having a fetus with trisomy 18 changed little, 1.9% (95% confidence interval = 0.4 to 5.5%). Our data, combined with those of the literature, suggest that the risk of finding an abnormal fetal karyotype in the presence of isolated choroid plexus cysts is at least 1% and may be as high as 2.4%. On the basis of these results, genetic counseling and prenatal diagnosis should be offered to these patients. PMID- 8731448 TI - Effect of indomethacin on individual amniotic fluid indices in multiple gestations. AB - To determine the effect of indomethacin on the amniotic fluid index of individual fetuses in multiple gestations, we evaluated seven sets of twins and two sets of triplets in preterm labor (eight cases) or with polyhydramnios (one case). The dose of oral indomethacin was either 25 mg every 12 hours or 25 mg every 6 hours. The amniotic fluid volume of individual amniotic sacs was estimated serially by the amniotic fluid index. When an oral dose of indomethacin of 25 mg every 6 hours was used, oligohydramnios eventually occurred in five of eight amniotic sacs and a subjective decrease in amniotic fluid occurred in one sac. Oligohydramnios was detected in only one of 13 amniotic sacs when the dose of indomethacin was 25 mg every 12 hours. The amniotic fluid volume returned to normal in these seven amniotic sacs within 4 days of discontinuing indomethacin. We conclude that maternal indomethacin therapy has a variable effect on the individual amniotic fluid volumes in multiple gestations. PMID- 8731449 TI - Safety of transvaginal fine needle puncture of gynecologic masses: a report after 500 consecutive procedures. AB - Transvaginal sonographically guided puncture may be used for diagnostic and therapeutic purpose in general gynecology and in oncology. To define the complication rate of this technique, 500 consecutive procedures were reviewed; 289 procedures were on cystic masses, 146 on solid tumors, 34 on mixed tumors, and 31 on other lesions. No life-threatening complications occurred. Early mild and self-limited complications developed in eight procedures (1.6%). Short-term complications occurred in four cases (0.8%; two infections, one pelvic pain, one hemorrhage); two of these required surgery. Transvaginal sonographically guided puncture is safe and does not entail a higher risk of infection than transabdominal punctures. PMID- 8731450 TI - "Starry sky" liver with fasting: variations in glycogen stores? PMID- 8731451 TI - Unusual course of the gonadal vein: a case report of postpartum ovarian vein thrombosis mimicking acute appendicitis clinically and sonographically. PMID- 8731452 TI - Dilated subarachnoid cisterna ambiens: a potential sonographic sign predicting cerebellar hypoplasia. PMID- 8731453 TI - Antenatal diagnosis of esophageal atresia with tracheoesophageal fistula. PMID- 8731454 TI - Linkage analysis of families with bipolar illness and chromosome 18 markers. AB - Linkage of bipolar (BP) illness with chromosome 18 markers located at 18p11 was recently reported. A possible role for chromosome 18 in the etiology of BP illness was implicated previously by the finding in three unrelated patients of a ring chromosome with breakpoints and deleted segments at 18pter-p11 and 18q23 qter. To test the potential importance of a gene defect on chromosome 18 in our material, we examined linkage with chromosome 18 markers in two families with multiple patients with BP illness or BP spectrum disorders. fourteen simple tandem repeat polymorphisms were used located in the chromosomal region 18p11 to 18q23 and separated by distances of approximately 10 cM on the genetic map. In one family linkage to chromosome 18 could not be excluded. Linkage and segregation analysis in the family suggests that the 12-cM region between D18S51 and D18S61 located at 18q21.33-q23 may contain a candidate gene for BP illness. PMID- 8731455 TI - Analysis of chromosome 18 DNA markers in multiplex pedigrees with manic depression. AB - Six pedigrees segregating manic-depressive illness (MDI) were analyzed for linkage to 21 highly polymorphic microsatellite DNA markers on chromosome 18. These markers span almost the entire length of the chromosome, and gaps between markers are less than 20 cM. In particular, we analyzed several markers localizing to the pericentromeric region of chromosome 18 which generated lod scores suggestive of linkage in an independent study. Lod score analysis was performed and results were examined by family. One region produced positive lod scores, though at 18q23 and not in the pericentromeric region. We additionally used two nonparametric methods because the true mode of transmission of MDI is unknown; results were again somewhat suggestive for markers in the region of 18q23 but not in the pericentromeric region. PMID- 8731456 TI - Platelet phosphoinositide signaling system: an overstimulated pathway in depression. AB - In order to test a possible depression-associated defect in signal transduction, platelet alpha 2-adrenergic-mediated phosphoinositide (PI) hydrolysis was measured, both in drug-free major depressed patients and in control healthy subjects. Results that express phospholipase C activity have shown significant increase in the metabolites of epinephrine-stimulated tritiated phosphatidyl-4,5 biphosphate (3H-PIP2) with respect to basal activity (saline-stimulated). Thrombin (2 units) and 10 mM sodium fluoride (NaF) also induced an increase in 3H PIP2 metabolites. These increases were potentiated in drug-free depressed patients both in epinephrine-and thrombin-stimulated platelets. In contrast, sodium fluoride, which directly stimulates G protein without receptor interaction, did not differentiate between patients and controls with respect to PI hydrolysis. This result suggests a possible depression-associated defect in heterologous receptor-G protein interaction. PMID- 8731457 TI - Cerebrospinal fluid corticotropin-releasing factor concentrations in patients with anxiety disorders and normal comparison subjects. AB - Cerebrospinal fluid (CSF) concentrations of corticotropin-releasing factor (CRF) were measured in a group of patients with anxiety disorders and normal comparison subjects (NC) to explore the hypothesis that abnormalities in CRF neuronal regulation occur in patients with anxiety disorders. Analysis of variance (ANOVA) revealed no differences in CSF CRF concentrations between the four diagnostic categories: panic disorder (PD), generalized anxiety disorder (GAD), obsessive compulsive disorder (OCD), and NCs. Male OCD patients had higher CSF CRF concentrations than men with PD and GAD and male NCs. CSF CRF concentration was positively correlated with age in women but not in men. These findings suggest that central neuronal CRF regulation may be affected by both age and gender. PMID- 8731458 TI - Serotonin-induced increases in platelet cytosolic calcium concentration in depressed, schizophrenic, and substance abuse patients. AB - Exaggerated intracellular calcium responses to challenges with serotonin (5-HT) have been reported in depression. In our studies, consistent with previous reports, patients with depression exhibited an exaggerated increase in 5-HT stimulated intracellular calcium concentration ([Ca++]i). Basal cytosolic calcium was elevated in both calcium-free and 1 mM calcium media in depressed patients. the increased responsiveness to 5-HT was seen in both conditions. Patients with schizophrenia and substance abuse did not differ from normal controls. The 5-HT response was correlated with diastolic blood pressure (r = 0.33, p = 0.02): however, this association did not fully account for the exaggerated [Ca++]i responses in the depressed group. These findings suggest that exaggerated increases in [Ca++]i in response to serotonin are a characteristic of depressed patients not shared with schizophrenic and substance abuse patients. The relationship of depression to hypertension, two conditions that share abnormalities of calcium homeostasis, warrants further study. PMID- 8731459 TI - Premature disinhibition of P3 generation in schizophrenia. AB - P300 (P3) is a long-latency cognitive event-related potential (ERP) elicited by relevant target stimuli. P3 was recorded from 11 schizophrenics and 13 normal controls during a cued visual continuous performance task (CPT-AX). Cue-target sequences were presented at short and long interstimulus intervals (ISIs), in order to investigate working memory in schizophrenia. There was no significant between-group difference in P3 amplitude to validly or invalidly cued targets at short ISI. In contrast, P3 amplitude to invalidly cued targets at long ISI was significantly greater in schizophrenics than in controls, suggesting decreased ability to encode or maintain inhibitory representations of stimulus context. P3 amplitude is typically reduced in schizophrenic subjects in the auditory modality, and normal or reduced in the visual modality. This study, which demonstrates a paradoxical P3 increase to targets at long ISI, suggests that P3 impairment in schizophrenia cannot be attributed solely to structural deficits within P3-generator regions. PMID- 8731460 TI - Electrophysiological findings during the menstrual cycle in women with and without late luteal phase dysphoric disorder: relationship to risk for alcoholism? AB - This study evaluated electrophysiological (EEG, ERGs), and cognitive (neuropsychological testing) responses in patients with late luteal phase dysphoric disorder (LLPDD, DSM-III-R) and controls over the menstrual cycle. In both groups, the frequency and stability of electroencephalogram (EEG) alpha activity significantly differed over the menstrual cycle. The latency of the P3 components of the auditory event-related potentials (ERPs) did not vary as a function of the menstrual cycle, but the P3 latency was found to be later in LLPDD subjects as a group. When the LLPDD subjects were assessed based on family history of alcoholism, it was found that those with alcoholic relatives had more high-frequency alpha (9-12 Hz) in their EEG, lower P3 component amplitudes, and longer P3 component latencies when compared to LLPDD subjects without alcoholic relatives or controls. These data suggest that LLPDD may have persistent neurophysiological correlates, some of which are also in common with risk for alcoholism. PMID- 8731461 TI - Alternation learning in obsessive-compulsive disorder. AB - The Wisconsin Card Sorting Test (WCST) and an alternation learning task were administered to 15 women with obsessive-compulsive disorder (OCD) and 15 age-, sex-, education-, and intelligence-matched healthy controls. OCD patients were significantly slower on the WCST as compared to the controls. Their performance on the alternation learning task was impaired relative to the control group, though this difference was diminished when we used education as a covariate. We found a significant positive correlation between performance on the alternation task and severity of symptoms in the OCD group. Performance of similar alternation tasks is impaired by damage to the orbitofrontal cortex in nonhuman primates. Therefore the data presented support the hypothesis of orbitofrontal cortex dysfunction in OCD. PMID- 8731462 TI - Site and size of lesion and psychosurgical outcome in obsessive-compulsive disorder: a magnetic resonance imaging study. PMID- 8731463 TI - Amplitude and area of the auditory P300 recorded with eyes open reflect remission of schizophrenia. PMID- 8731464 TI - Neuropsychological functioning in trichotillomania. PMID- 8731465 TI - Recall performance in acutely depressed patients and plasma cortisol. PMID- 8731466 TI - Population prevalence of psychopathology in children and adolescents with intellectual disability: I. Rationale and methods. AB - Our knowledge of the epidemiology of psychopathology in children and adolescents with intellectual disability (ID) is hampered by a number of factors. These include the relative scarcity of studies of children rather than adults, the study of non-epidemiological samples such as those in institutions or those attending psychiatric clinics, a lack of standardized methods of assessment of psychopathology, studies with numbers too small to provide adequate confidence intervals around identified prevalence rates, insufficient detail concerning symptoms or syndromes as well as disorder, and a lack of consistent data concerning the effects of basic demographic variables such as age, sex and IQ. Despite these limitations, it is clear that psychopathology is several times more prevalent in children and adolescents with ID than in those without this disability. This paper reviews findings from previous studies and describes the methodology of a new study using the Developmental Behaviour Checklist. PMID- 8731467 TI - Population prevalence of psychopathology in children and adolescents with intellectual disability: II. Epidemiological findings. AB - This paper reports findings from an epidemiologically derived population in a multicentre study in NSW, Australia. The design of this study is described in the accompanying paper. Those with mild intellectual disability (ID) were likely to have been underascertained, but identification and participation rates for those with more severe ID were high. The study found that in the regions surveyed 40.7% of those with ID and aged between 4 and 18 could be classified as having severe emotional and behaviour disorder or as being psychiatrically disordered. The profoundly intellectually handicapped had lower levels of disturbance overall compared with those with mild, moderate and severe ID. The level of ID affected scores on a number of behavioural dimensions, with disruptive and antisocial behaviours more prominent in the mild ID group, and 'self-absorbed' and 'autistic' behaviours more prominent in those with severe ID. Age and sex did not affect prevalence, a finding that is in contrast to that found in general child psychopathology. The study found that fewer than 10% of children with intellectual disability and major psychiatric disorder had received specialist assistance. PMID- 8731468 TI - Piagetian principles: simple and effective application. AB - A simplified version of Piaget's sensorimotor levels was employed to teach a group of 25 extremely impaired individuals who live in a large residential facility. Throughout the facility, all available formal assessments using the Vineland Adaptive Behaviour Scales (VABS) were compared. Using this tool, an average of 13% of individuals from the common population pool increased their scores over a 6-year time period. All of the individuals who received training within the experimental group demonstrated increased scores. Scores increased such that an average gain of nearly 1 month in overall age equivalency per individual per year was realized. A matched group comparison, a prediction test for like sensorimotor skill attainment (the primary distinction of this curriculum methodology), and an historical review of subject skill training, all support the cognitively geared methodology as being primarily responsible for this accelerated progress. PMID- 8731469 TI - The social and community involvement of older Australians with intellectual disabilities. AB - This paper describes the involvement of older people with intellectual disabilities in residential- and community-based activities and programmes. The 446 participants were selected from a national database of people of 55 years of age and over with intellectual disabilities in Australia. The data indicate that participants made infrequent use of public amenities and social and recreational facilities. With the exception of those living with relatives, most had limited contact with family and friends. The discussion draws attention to the need for independence training which will enable the current generation of middle-aged people with an intellectual disability to make decisions and choices about social and community activities. PMID- 8731470 TI - A behavioural approach to the treatment of non-retentive encopresis in adults with learning disabilities. AB - Problems such as encopresis are perceived as "normal' for some populations. For example, within the field of learning disability, little serious study has been devoted to either prevalence or aetiology of encopresis. Treatment issues have been obscured by problems of definition and aetiology. Treatment reports are rare despite the apparent magnitude of the problem, and involve a disproportionate number of cases with a mild intellectual disability or secondary encopresis. The following report describes a reinforcement-based treatment programme for primary, non-retentive encopresis in five young adult men with mainly more severe learning disabilities. Major soiling accidents were eliminated in four out of five cases and substantially reduced in the fifth. Treatment times were long. Issues relating to the use of aversive techniques are discussed, as are the limitations of the present study. PMID- 8731471 TI - Vision care requirements among intellectually disabled adults: a residence-based pilot study. AB - Intellectually disabled adults have an high incidence of visual problems and they are often unable to communicate their visual difficulties. At Lennox Castle and Waverley Park Hospitals, vision care is through referral by medical and nursing staff to designated optometrists and ophthalmologists. This practice has provided a good service when visual difficulties are noticed. The vision care requirements of all residents had not been comprehensively assessed and a new interdisciplinary procedure developed at Waverley Park Hospital had drawn attention to the fact that only 11% had been offered vision assessment within the previous 5 years. In this study, 63 residents without specific referral received a comprehensive visual and medical assessment. The residents ranged from age 20 to 85 years and included the full range of disabilities. Objective assessments, ophthalmoscopy and retinoscopy were generally successful for all levels of disability. Visual acuity and visual fields were tested using methods suitable for nonverbal subjects. Success rates for these subjects were generally good, except in the profoundly disabled group where less than 30% were able to respond. A high prevalence of visual impairment, refractive error, squint and other ocular conditions was found. Visual impairment was most common in the severe and profoundly disabled groups because of optic nerve or cortical dysfunction. New spectacles were recommended for 23 residents (seven others had adequate correction). Nine residents were referred for ophthalmologic consultation, mainly for cataract. Three required monitoring for visual conditions. Thirty-one residents (49%) required no immediate action beyond documentation of the visual status. This study has shown a high prevalence of visual difficulties which were not previously detected. Routine biennial vision assessment of all residents is recommended to allow timely intervention to correct vision problems, and also to provide the necessary information about vision to plan appropriate programmes of activity. PMID- 8731472 TI - The relevance of Vygotsky's theory of the "zone of proximal development' to the assessment of children with intellectual disabilities. AB - This paper reports a study which investigated the relevance of Vygotsky's concept of the "zone of proximal development' (ZPD) to the assessment of children with intellectual disabilities. The ZPD is the difference between a child's actual level of development shown by unassisted performance, and his or her potential level as indicated by assisted performance. This study aimed to test the validity of measuring the ZPD both among children with intellectual disabilities and in the area of map use. The results are discussed in terms of their bearing on the issues of assessment, instruction and the concept of intellectual disability. PMID- 8731473 TI - Maladaptive behaviour in Prader-Willi syndrome in adult life. AB - Thirty adults with Prader-Willi syndrome (PWS) were compared with 30 adults with non-specific learning disability matched for age, sex and severity of mental retardation. Maladaptive behaviour was assessed with the Aberrant Behavior Checklist (ABC), a 58-item structured interview which rates behaviours from 0 (not a problem) to 3 (severe problem) and which yields five factors (I) irritability, agitation; (II) lethargy, withdrawal; (III) stereotypic behavior; (IV) hyperactivity, non-compliance; and (V) inappropriate speech). The PWS sample had significantly higher factor I (P < 0.001) and factor V (P < 0.05) scores. The PWS sample had mean scores above 1 for 17 ABC items; the contrast subjects had no mean scores above 1. The factor I scores for the PWS sample were similar to those of inpatients in hospital facilities for adults with mental retardation and mental illness or severely challenging behaviour. The results support previous work, and extend it by suggesting that temper tantrums, self-injury, impulsiveness, lability of mood, inactivity and repetitive speech are characteristic behaviours in PWS in adult life. Studies of the reasons for heterogeneity in behaviour are now needed. PMID- 8731474 TI - Staff strategies and explanations for intervening with challenging behaviours. AB - Behavioural models identify the actions of others as an important factor in the development and maintenance of challenging behaviours. In the present study, 109 care staff were asked about their immediate and longer-term intervention strategies for a fictitious young man's challenging behaviour. Staff descriptions of long-term interventions were largely consistent with the aims of psychological interventions. However, their immediate intervention strategies were similar to the counter-habilitative strategies identified in previous observational and self report research. The implications of these results for models of staff behaviour, staff training, the design of behavioural programmes and future research are discussed. PMID- 8731475 TI - Composite versus multiple-rating scales in the assessment of psychopathology in people with mental retardation. AB - This study tested the hypothesis that separate ratings of frequency and duration add significant information to ratings of problem severity in the assessment of psychopathology in people with mental retardation. The Reiss Screen was modified to require ratings of problem severity, frequency and duration for each of 38 maladaptive behaviours. The 171 adolescents and adults were rated by caretakers and supervisory staff on the modified Reiss Screen. The three ratings were found to be very highly intercorrelated. Moreover, multiple linear regression analyses revealed that the ratings of frequency and duration did not add significant information to that provided by ratings of problem severity in the prediction of psychiatric diagnosis in the person's case file. The results support the use of composite scaling over multiple-ratings in the assessment of psychopathology in persons with mental retardation. PMID- 8731476 TI - Life expectancy of mentally retarded hemiplegics. AB - Sixty-four mentally retarded people with hemiplegia (35 females and 29 males), first recorded in 1963, were re-examined 30 years later for life expectancy. Detailed physical and mental states, lengths of hospital stays and other information were noted. Recent advances in diagnosis and prognosis of hemiplegics were included. The results of the study indicate that, with special provisions available, people with hemiplegia have the prospect of reaching pensionable age and beyond: the oldest female is 85 and the oldest male 76 years of age. PMID- 8731477 TI - Community care for adults with learning disability and their carers: needs and outcomes from the Leicestershire register. AB - A cross-sectional study, based on an epidemiological register, was carried out to describe the prevalence of disabilities, felt needs and use of services for adults with learning disability and to compare outcomes of reported morbidity, stress and satisfaction among their informal carers. Subjects included 2117 adults and 982 carers known to specialist services in Leicestershire. Behavioural and psychological problems and epilepsy were the main disabilities in adults. The leading unmet needs reported by residential carers were for daycare and other forms of residence, and those reported by informal carers were for financial help, long-term social support, respite care and housing adaptations. Informal carers reported 40% more limiting health disorders compared to the general population, with depression almost four times more common among female carers. Divisions between health and social care are causing inequality and hardship. Lifelong informal carers need options for independence. The increase over time in the prevalence of adults with severe learning disabilities adds to the evidence that more resources for care are needed. Epidemiological registers and methods should be developed to aid purchasing and provision for this client group. PMID- 8731478 TI - Myelin-associated glycoprotein inhibits axonal regeneration from a variety of neurons via interaction with a sialoglycoprotein. AB - Myelin-associated glycoprotein (MAG) is a potent inhibitor of axonal regeneration from both cerebellar neurons and adult dorsal root ganglion (DRG) neurons. In contrast, MAG promotes axonal growth from newborn DRG neurons. Here, we show that the switch in response to MAG from promotion to inhibition of neurite outgrowth by DRg neurons occurs sharply at Postnatal Day 3. To date, of all the neurons tested a postnatal switch in response is only observed for DRG neurons; MAG inhibits axonal growth from retinal, superior cervical ganglion, spinal, and hippocampal neurons of all postnatal ages. Furthermore, MAG binds to neurons from which it promotes and from which it inhibits outgrowth, in a sialic-acid dependent manner. Now we show this binding is also trypsin-sensitive. Hence, the interaction is via a sialoglycoprotein. Binding of MAG to all the neurons tested here was also sialic-acid-dependent. Importantly, both inhibition and promotion of neurite outgrowth by MAG are reduced, or abolished completely, either by desialyation of the neurons prior to the outgrowth assay or by including small sialic-acid-bearing sugars in the cultures. These results suggest that MAG is likely to contribute to the lack of regeneration observed throughout the nervous system. Also, it is likely that MAG is exerting its effect, either directly or indirectly, on both promotion and inhibition of neurite outgrowth via a neuronal sialoglycoprotein. PMID- 8731479 TI - Glutamate receptor ion channel properties predict vulnerability to cytotoxicity in a transfected nonneuronal cell line. AB - Excessive activation of glutamate receptors is thought to play a critical role in neuronal excitotoxicity. To compare the cytotoxic potential of different glutamate receptor subtypes and correlate receptor biophysical properties with cytotoxicity, we have expressed recombinant receptors in human embryonic kidney 293 (HEK-293) cells. Survival of transfected cells was analyzed under conditions of defined agonist concentration and exposure time. For HEK-293 cells transfected with N-methyl-D-aspartate (NMDA) receptors, the EC50 for NMDA-induced cytotoxicity was 300 microM. Experiments using ion substitution, or cells expressing mutant NMDA receptors with low calcium permeability, suggested that both calcium and sodium influx through NMDA receptors contributed to cytotoxicity. In contrast, cytotoxicity was not observed in cells transfected with calcium permeable alpha-amino 3-hydroxy-5-methyl-4-isoxazole propionate- or kainate-type glutamate receptors even at saturating agonist concentrations, unless inhibitors of agonist-dependent desensitization were included. These results directly demonstrate that calcium permeability and desensitization kinetics play important roles in determining the excitotoxic potential of different glutamate receptor subtypes. PMID- 8731480 TI - Differential mRNA transport and the regulation of protein synthesis: selective sensitivity of Purkinje cell dendritic mRNAs to translational inhibition. AB - Although the majority of mRNAs expressed in neurons are confined to the perikaryon, a growing number appear to be transported into dendrites. It is likely that this allows for the local regulation of protein synthesis within discrete subcellular compartments. Here, three different subcellular distribution patterns are demonstrated for four mRNAs that encode proteins highly expressed in Purkinje cells and their dendrites; mRNAs are found in the perikaryon only, perikaryon and proximal dendrite, or perikaryon and proximal plus distal dendrites. Further, it is shown that transport of an mRNA into the dendrites increases its sensitivity to translational inhibition by diphtheria toxin. These data suggest a simple model whereby the transport machinery can regulate the translation of selected mRNAs. Thus, environmental signals that generally affect translational efficiency in concert with the selectivity provided by the transport machinery could provide a means to locally regulate the synthesis of a restricted pool of proteins. PMID- 8731481 TI - Nerve growth factor regulates the expression of brain-derived neurotrophic factor mRNA in the peripheral nervous system. AB - Neurotrophins are profound regulators of neuronal survival in the developing peripheral nervous system and are synthesized by peripheral neurons themselves both during development and in maturity. Neuronal neurotrophin expression may be importantly related to survival of mature neurons, both in normal and pathological states. We show here that brain-derived neurotrophic factor (BDNF) gene expression in dorsal root ganglia is strongly stimulated in vivo by another neurotrophin, nerve growth factor (NGF). Furthermore, colocalization studies show that many BDNF-expressing sensory neurons also express trk A, the high-affinity NGF receptor. These results demonstrate a novel regulatory mechanism for neurotrophin gene expression and suggest a paracrine function for neurotrophins in mature animals. PMID- 8731482 TI - Paracrine interactions of BDNF involving NGF-dependent embryonic sensory neurons. AB - The expression of BDNF mRNA by a proportion of embryonic dorsal root ganglion neurons has led to the proposal that BDNF acts by an autocrine loop on these neurons. To clarify the role of BDNF expression in developing sensory neurons, we measured the level of BDNF mRNA in purified populations of cranial sensory neurons that depend on either NGF or BDNF for survival. When neuronal death is taking place, the highest levels of BDNF mRNA were detected in NGF-dependent cutaneous sensory neurons. BDNF mRNA was expressed at lower levels in BDNF dependent cutaneous sensory neurons and was undetectable in BDNF-dependent proprioceptive neurons. In coculture, NGF-dependent neurons promoted the survival of BDNF-dependent neurons by the production and release of BDNF. Depolarizing levels of KCl increased the expression of BDNF mRNA in cultured sensory neurons and this effect was partially inhibited by calcium channel antagonists. Our results suggest that during the phase of naturally occurring neuronal death, BDNF acts by a paracrine mechanism in sensory neurons and that BDNF expression is regulated by neural activity. PMID- 8731483 TI - Signals triggering the induction of leukemia inhibitory factor in sympathetic superior cervical ganglia and their nerve trunks after axonal injury. AB - Leukemia inhibitory factor (LIF) plays an important role in regulating neuropeptide expression in sympathetic and sensory neurons after axonal transection. By 2 h after axotomy, LIF mRNA increased in nonneuronal cells in sympathetic ganglia and peripheral nerve. In addition, within 1 h of explanting sympathetic ganglia or segments of sympathetic nerve trunks, a protein factor(s) that was able to induce LIF mRNA both in sympathetic cultures and in intact ganglia in vivo was released. This factor(s) appeared to be present in sympathetic ganglia and their nerve trunks under normal conditions and to be activated and/or released after axonal injury. Since the factor(s) has a molecular weight(s) greater than 66 kDa, and no other proteins of such high molecular weight have been previously identified with LIF-inducing activity, it appears to be a novel inducer of LIF. PMID- 8731484 TI - Prevention of prematurity: important and difficult. PMID- 8731485 TI - Advances in perinatology. PMID- 8731486 TI - Prevention of prematurity. PMID- 8731487 TI - Intrapartum fetal surveillance: a reappraisal. PMID- 8731489 TI - Common clinical manifestations of maternal diabetes in newborn infants: implications for the practicing pediatrician. AB - Even though perinatal mortality of infants of diabetic mothers has decreased remarkably in recent years and now approaches that of the general population, these infants still face a multitude of potential complications and the propensity for increased morbidity, both in utero and postnatally. Many of these complications are clearly related to the metabolic status of the diabetic mother. Increasing awareness among insulin-dependent diabetic patients and health providers of the need for glycemic control and the ever-growing understanding of the peculiarities of diabetic pregnancies eventually should combine to provide the best possible outcome for these infants. PMID- 8731488 TI - Group B streptococcus and pregnancy. AB - Group B streptococcus is a significant pathogen for both mother and child. routine urine culture in pregnancy will identify and allow treatment of women with asymptomatic bacteriuria. An optimal protocol for the prevention of neonatal sepsis has not yet been developed. While intrapartum antimicrobial prophylaxis appears to provide the best potential, each of the currently suggested protocols has significant drawbacks. Drawbacks include the potential for missing high-risk carriers, failure to treat a significant proportion of those destined to deliver an affected infant because no risk factors are present, and empirical treatment of a large proportion of the population in order to present significant disease in a few. Until an effective program of immunization becomes available, intrapartum prophylaxis of group B streptococcal carriers appears to offer the best hope of reducing the incidence of neonatal disease. Caregivers should adopt a uniform practice with regard to screening and prophylaxis. It is essential that any broad-based screening program include an evaluation of efficacy as well as complications including the development of new etiologic agents as causes of neonatal sepsis and the emergence of resistant bacteria. Further, mothers and newborns should be evaluated for drug adverse reactions and the impact of intrapartum prophylaxis on the use of prolonged empirical broad-spectrum antimicrobial therapy on the asymptomatic infant. Additional research is necessary regarding the required duration of therapy for optimal effect of intrapartum prophylaxis, the need for postnatal prophylaxis of the asymptomatic neonate, and the optimal agent for neonatal prophylaxis (penicillin versus broad spectrum agents) if neonatal therapy is necessary after intrapartum prophylaxis. PMID- 8731490 TI - The clinical significance of asymmetric intrauterine growth retardation. PMID- 8731491 TI - Ultrasound detection of fetal abnormalities. PMID- 8731492 TI - Managing fatigue in operational settings. 1: Physiological considerations and countermeasures. AB - The authors consider three aspects of managing fatigue in the workplace. They provide a brief overview of important scientific findings related to sleep and circadian physiology that establish the psychobiological foundation of fatigue. Their major focus is on the relevance of these findings to operational settings. In addition, they provide examples to describe practical fatigue countermeasures that can be used in operational settings. PMID- 8731493 TI - Managing fatigue in operational settings 2: An integrated approach. AB - The six domains that must be addressed in managing fatigue in operational settings are identified, and examples of how the aviation industry is dealing with the problems in each domain are given. Challenges facing healthcare providers in managing fatigue are also discussed. PMID- 8731494 TI - Methods of testing for sleepiness [corrected]. AB - Normal nonrandom fluctuation in daily human performance have been documented for years. Published research reports have shown patterns of workers' errors in reading gas meters, operators' delays in answering calls, drivers' drowsiness, sleepy locomotive engineers' automatic breaking, vehicle crashes, deaths resulting from disease, brief periods of sleep, and sleep latency in structured naps. The authors summarized these data sets and fitted them with a two-peak-per day cosine curve derived from the population growth function used in chaos theory. Median parameters extracted from the curve fits predicted a sharp peak of sleepiness at 2:30 AM and a secondary peak at 2:30 PM. The shape of the curve was modified by a nonlinear sleep-deprivation factor. The model appeared to be biological rather than behavioral or social because it applied well to disease related deaths. The authors also review measurement of sleepiness through electroencephalographic monitoring, self-reports, pupillography, and the Multiple Sleep Latency and the Maintenance of Wakefulness Tests. PMID- 8731495 TI - Some practical considerations and policy implications of studies of sleep patterns. PMID- 8731496 TI - Effect of chronic aerobic exercise and progressive relaxation on motor performance and affect following acute stress. AB - The effects of a 10-week aerobic exercise and progressive relaxation training program on somatic, emotional, and behavioral responses to acute stress, as determined by quality of motor performance and affect, were examined. The participants consisted of 60 unfit male university undergraduate students with no previous training in stress management who were randomly and evenly assigned to engage in one of four treatments over 10 weeks: (a) moderate aerobic exercise, (b) progressive relaxation, (c) a placebo group that engaged in group discussion but did experience acute stress, and (d) a nonintervention control group that did not experience stress while performing the motor task. Acute stress consisted of "losing" against a competitor of the opposite sex on the criterion motor task while receiving unpleasant information about their performance over 30 preintervention and 30 postintervention trials. Analyses indicated that aerobic exercisers, in comparisons with the other groups, responded to acute stress with more positive affect, lower stressor task heart rate, reduced systolic (but not diastolic) blood pressure, and superior motor performance. Progressive relaxation markedly reduced systolic blood pressure but did not favorably influence performance or affect in response to acute stress. Placebo and control groups were statistically similar on all measures. The findings indicated support for the use of chronic aerobic exercise as a strategy for improved coping with acute stress. PMID- 8731497 TI - Measurement of soluble thrombomodulin in sera from various clinical stages of diabetic nephropathy. AB - The levels of soluble thrombomodulin (TM) in serum samples were measured by one step sandwich enzyme immunoassay. The aim of the present study was to determine if levels of soluble TM in sera might correlate with disease activity in patients with diabetic nephropathy. Three hundred and twenty patients with diabetic nephropathy were examined. Patients with diabetic retinopathy were excluded from the present study. This study showed an increase of soluble TM levels in sera from patients with diabetic nephropathy. The levels of soluble TM in sera from the macroalbuminuric stage with renal dysfunction were significantly increased compared with those from the normo-, micro-, or macroalbuminuric stage of diabetic nephropathy without renal dysfunction. The increase of soluble TM in sera paralleled levels of urinary albumin, blood urea nitrogen (BUN), s creatinine (Cr), and duration of noninsulin-dependent diabetes mellitus (NIDDM). Furthermore, a decrease of TM staining in the glomerular capillary walls was observed in both microalbuminuric and macroalbuminuric stages by immunofluorescence. It appears that the measurement of soluble TM in sera is useful in evaluating the degree of glomerular endothelial injuries in patients with diabetic nephropathy. PMID- 8731498 TI - A new, fast, and simple DNA extraction method for HLA and VNTR genotyping by PCR amplification. AB - In the present study a new DNA extraction method is described. The new protocol, which uses caprylic acid for isolating DNA, is technically simple and very fast, as it enables us to obtain DNA from peripheral blood in only 10 minutes. Moreover, DNA preparations obtained with this procedure can be effectively used for HLA class II and variable number tandem repeat genotyping by polymerase chain reaction, so the new method is well suited for routine clinical use in any type of analysis requiring DNA typing for individual characterization. PMID- 8731499 TI - Detection of ciprofloxacin resistance mutations in Campylobacter jejuni gyrA by nonradioisotopic single-strand conformation polymorphism and direct DNA sequencing. AB - A total of 27 strains of Campylobacter jejuni (24 clinical strains and three laboratory strains) were examined for the presence of point mutations in the quinolone resistance determining region (QRDR) of gyrA gene by nonradioisotopic single-strand conformation polymorphism (non-RI SSCP) analysis with silver stain. Direct DNA sequencing of the polymerase chain reaction (PCR)-amplified DNA fragments confirmed the results obtained by non-RI SSCP analysis and revealed that in clinical strains high-level quinolone resistance [minimal inhibitory concentration (MIC) to ciprofloxacin > or = 16 micrograms/ml] was closely associated with one type of single-point mutation at codon 86 (Thr-Ile). Two strains with MICs of 8 and 1 microgram/ml showed point mutations at codons 86 and 70, respectively. Furthermore, transitions at codon 119 of the gyrA QRDR were identified in 17 strains. Six types of bands were separated in a single electrophoretic step with silver stain within 2 hours after PCR amplification of the gyrA QRDR as follows: type I associated to mutation at codon 70 (Ala-Thr), type II to mutation at codon 90 (Asp-Asn), type III to variant with transition at 119, type IV to wild-type, type V to mutation at codon 86 (Thr-Ile), and type VI to mutation at codon 86 (Thr-Ile) and transition at codon 119. Using four DNA extracts from Cambylobacter coli organisms as templates for amplification of the gyrA QRDR, no PCR products were obtained. Non-RI SSCP was proved to be a simple, rapid, and useful screening method for detecting gyrA mutations associated with ciprofloxacin resistance in C. jejuni. PMID- 8731500 TI - Effects of storage on in vitro platelet responses: comparison of ACD and Na citrate anticoagulated samples. AB - The present study was designed to evaluate whether the use of acid citrate dextrose (ACD) Formula A may enhance the survival of platelets during storage, thus allowing the continuance of platelet studies over the period of 2-3 hours usually recommended. For this purpose the effects of time on in vitro platelet response to several agonists have been investigated in platelet-rich plasma (PRP) obtained from blood samples anticoagulated with either Na citrate or ACD Formula A. The analysis of the data obtained in in vitro platelet aggregation studies using various parameters and at different time points demonstrated that storage of PRP obtained from citrated samples caused a marked reduction of platelet responses. This reduction was already evident after 6 hours, and a strong decrease was observed after 8 hours with all the agonists used. On the other hand, storage of ACD anticoagulated blood did not cause any significant decrease of platelet responsiveness up to 6 hours. A reduction of platelet aggregation became evident only after 8 hours, but not to the same extent as the one observed in citrated samples. Therefore, it may be concluded that the use of ACD Formula A as anticoagulant is capable of maintaining a normal platelet responsiveness up to 6-8 hours, thus permitting the investigation of platelet function for periods of time over those commonly recommended. PMID- 8731501 TI - Antigen association of total HEp-2 cell extract in the screening of autoimmune diseases by western blot. AB - Sera from patients with autoimmune diseases and healthy individuals were comparatively analyzed by Western blot, using total extract from HEp-2 cells as antigen. Bands between 14 and 20 kDa were present in 99% of patients with autoimmune diseases and symptomatic seronegative patients, in contrast to only 4% of healthy controls. No bands were detected in the remaining 96% of healthy controls. We propose these antigens as pathologically important for the diagnostic screening of patients with suspected autoimmune disease. PMID- 8731502 TI - Characterization of a monoclonal antibody specific for H type 2 structure of ABO blood group, and its use for measuring H type 2 on human red blood cells. AB - A monoclonal antibody 3A5 specific for the human red blood cells was produced by immunizing BALB/c mouse with human erythrocyte membranes of group O following the immunization protocol of selectively killing the antigen-stimulated lymphocytes. The monoclonal antibody 3A5 we obtained agglutinated red blood cells regardless of individuals of blood group A, B, or O types, but not those from a person with a rare para-Bombay type which is the H-deficient phenotype. The hemagglutination reaction of 3A5 was not inhibited by saliva from either secretor or nonsecretor individuals. The specificity of 3A5 was studied by adsorption with and elution from synthetic oligosaccharide immunoadsorbents including H disaccharide, and H type 1, 2, 3, and 4 structures. Also, the reactivity of 3A5 with synthetic oligosaccharide-BSA complexes for H type 1, H type 2, Le(a), Le(b), A, and B was determined by an immunoassay. We found that 3A5 did not react with any of these synthetic oligosaccharides except H type 2. From these results, the antigenic epitope recognized by 3A5 was demonstrated to be the H type 2 structure. Additionally, the H type 2 substance on erythrocytes was quantitatively analyzed using 3A5. PMID- 8731503 TI - Detection of salivary and seric IgG and IgA antipooled pigeon sera activities in patients with pigeon breeder's disease. AB - Pigeon breeder's disease (PBD) is an interstitial lung disease induced by exposure to pigeon antigens. Search of antipigeon antigen antibodies (APSA) in serum or bronchoalveolar lavage is generally used for auxiliary diagnostic purposes. However, APSA can be present in a number of exposed but asymptomatic individuals as well as in patients with other interstitial lung diseases who live in areas where keeping pigeons is a common domestic habit. In this study, saliva was evaluated as an alternative means to serum for APSA detection by ELISA using pooled pigeon sera as antigen. Serum and saliva samples obtained from 17 patients with PBD, 14 with idiopathic pulmonary fibrosis (IPF), 19 asymptomatic relatives (AR) exposed to pigeon antigens, and 27 clinical healthy voluntary subjects (CHVS) were tested for IgG and IgA APSA. Our results showed that both fluids obtained from PBD patients exhibited a significantly higher specific IgG antibody activity compared to the other groups. Serum optical density (O.D.) values for PBD were 1.187 +/- 0.738 vs. 0.024 +/- 0.033, 0.255 +/- 0.471, and 0.204 +/- 0.346 for CHVS, AR and IPF, respectively (P < 0.05). Salivary O.D. for PBD were 0.801 +/- 0.447 vs 0.010 +/- 0.011, 0.104 +/- 0.151, and 0.22 +/- 0.447 (P < 0.05). In contrast, serum specific IgA did not discriminate between PBD and IPF patients. In addition, although the PBD group exhibited the highest values of IgA salivary APSA, high levels were also observed in saliva specimens from CHVS, a group of normal individuals who deny pigeon exposure. These findings suggest that measurement of IgG salivary APSA can play a role in the evaluation process of patients with pigeon breeder's disease. PMID- 8731504 TI - Technicon Immuno 1 PSA assay measures both free and alpha-1-antichymotrypsin complexed prostate-specific antigen on an equimolar basis. AB - In this study, we investigated the immunoreactivity of the Technicon Immuno 1 PSA assay, a monoclonal-polyclonal sandwich immunoassay, with free and ACT-complexed PSA. Assay calibrators prepared with free PSA (standard Immuno 1 calibrators) and calibrators consisting of 90% ACT-complexed and 10% free PSA (90:10 calibrators) yielded virtually identical PSA recoveries at all concentrations tested. Concentrations of total PSA at approximately 4 and 10 ng/mL, prepared in varying ratios of free PSA to PSA-ACT complex, recovered from 92-104% at 4 ng/mL and from 98-102% at 10 ng/mL. Additionally, an excellent correlation of serum total PSA values from a panel of 40 prostatic cancer patient samples was obtained using calibration curves generated with the standard Immuno 1 calibrators or the 90:10 calibrators. These results demonstrate that the Technicon Immuno 1 PSA assay measures free and ACT-complexed PSA on an equal molar basis. PMID- 8731505 TI - Antibody-interleukin 2 fusion proteins: a new approach to cancer therapy. PMID- 8731507 TI - Erythrocyte aggregation and erythrocyte deformability modify the permeability of erythrocyte enriched fibrin network. AB - Intravascular thrombus formed under low shear conditions consists of red cells enmeshed within a fibrin network. Since red cells reduce the permeability of fibrin network by surface drag and by volume occupancy the significance of red cell aggregability and deformability in network permeability needs examination. In this study networks were developed by the addition of thrombin to washed red cells suspended in platelet free plasma. The effects of the polymers polyvinylpyrrolidone (PVP) and poloxamer 188 on network permeability were compared to gauge the influence of red cell aggregation. Both polymers increase network permeability by an action on fibrin polymerisation but PVP alone enhances red cell aggregation. PVP was found to increase network permeability significantly both by increasing the permeability of the fibrin component of the network and by increasing red cell aggregation and thus reducing red cell surface drag. In separate experiments red cells were pre-treated with heat, glutaraldehyde, or diamide to reduce cell deformability. Decreased cell deformability caused significant reductions in network permeability. This was ascribed to the reduced aggregability of hardened red cells. Red cell aggregation during coagulation enhances molecular transport through modifying the network. This may have implications for the penetration of fibrinolytic agents. PMID- 8731506 TI - The contribution of thrombosis to the clinical expression of coronary atherosclerosis. PMID- 8731508 TI - Effects of H1 histones and a monoclonal autoantibody to H1 histones on clot formation in vitro: possible implications in the antiphospholipid syndrome. AB - Histones are known to bind anionic phospholipids (PLs). Binding of procoagulant PLs by histones released during cell injury/death may interfere with coagulation and may serve a local regulatory anticoagulant function. Histone H1 prolonged the PT and APTT of normal pooled plasma (NPP). These increased clotting times disappeared when anti-H1 monoclonal antibody (mAb) was added to the incubation. Dilute Russell Viper Venom Time was also prolonged with the addition of histone H1. When H1 was added to plasma from a patient with the antiphospholipid syndrome (APL plasma), there was a further prolongation of the abnormal APL clotting time which was partially corrected by anti-H1 mAb. Platelet neutralization times were increased with added H1 and were further increased using APL plasma. when disrupted endothelial cells were incubated with plasma with and without anti-H1 antibodies, the addition of anti-H1 antibodies decreased clotting times. These data support the theory that histones released during cell injury may have a regulatory anticoagulant role in clot formation and the anti-H1 effect of some APL plasmas may inhibit this, thereby contributing to thrombosis seen in APL patients. PMID- 8731509 TI - Increase in plasma thrombomodulin level in patients with vibration syndrome. AB - To determine whether endothelial cells are injured in vibration syndrome, we measured plasma levels of thrombomodulin (TM) in 100 patients with this syndrome using one-step sandwich enzyme immunoassay. Plasma level of TM in patients with vibration syndrome was significantly higher than that in normal control (p < 0.0001). There was no significant difference in the plasma TM level between patients with vibration syndrome and those with collagen disease. Plasma TM concentration in chain-saw operators was significantly higher than that in rock drill operators (p < 0.05). Plasma TM value did not significantly differ between patients with vibration-induced white finger (VWF) and those without VWF. These results suggest that endothelial injury is present in patients with vibration syndrome, the degree of endothelial injury in patients with vibration syndrome equals that in patients with collagen disease, and the endothelial injury in chain-saw operators is greater than that in rock-drill operators. However, there was no difference in the degree of endothelial injury between patients with VWF and those without VWF. PMID- 8731510 TI - Insulin exerts opposite effects on platelet function at physiological and supraphysiological concentrations. AB - In this study, we investigated the effects of a 3-min insulin incubation both at physiological and at supraphysiological concentrations on platelet aggregation and intraplatelet cyclic guanosine monophosphate (cGMP) levels both in the absence and in the presence of phosphodiesterase inhibition. We observed that insulin at concentration in the range 0.25-2 nmol/L decreases platelet response to adenosine 5-diphosphate (ADP), being Effective Dose 50 (ED50) for ADP with 2 nmol/L insulin 164 +/- 15% of the basal value, p = 0.005; furthermore, insulin increases intraplatelet content of cGMP (from basal 7.3 +/-0.6 pmol/10(9) plts to 14.6 +/- 1.2 pmol/10(9) plts with 2 nmol/L insulin, p=0.0001) and does not affect the platelet cGMP increase induced by nitrates. On the contrary, at very elevated concentrations (25-200 nmol/L) insulin increases platelet aggregation to ADP (ADP ED50 with 200 nmol/L insulin being 81 +/- 4% of the basal value, p = 0.01), decreases intraplatelet content of cGMP (from basal 7.2 +/- 0.1 pmol/10(9) plts to 5.7 +/- 0.2 pmol/10(9) plts with 200 nmol/L insulin, p = 0.01) and attenuates the platelet cGMP increase induced by nitrates. When cGMP catabolism is inhibited by theophylline or the selective cGMP phosphodiesterase inhibitor zaprinast, insulin shows anti-aggregating effects also at highly supraphysiological concentration (25-200 nmol/L). These results indicate that insulin, depending on the concentrations employed, shows opposite effects on platelet function, and they provide information about the mechanisms involved: actually, insulin is able to increase both cGMP synthesis, through guanylate cyclase activation, and cGMP catabolism, through phosphodiesterase activation. At physiological or slightly supraphysiological concentrations the first phenomenon is prevailing, so that cGMP intraplatelet values increase and insulin shows antiaggregating properties, whereas, at supraphysiological concentrations, insulin reduces cGMP levels through a prevailing phosphodiesterase activation, as supported by the fact that, when cGMP catabolism is prevented, insulin shows anti-aggregating properties also at the highest concentrations used. PMID- 8731511 TI - Morphological and hemostatic changes in rats with abdominal arterial prosthesis. AB - We evaluated the changes over time in hemostatic factors during ongoing arterial thrombosis in rats, as induced by a loop-shaped aortic prosthesis. Moreover, we investigated this condition by inspecting in parallel local thrombus growth, systemic vascular prostacyclin and t-PA production. One minute after loop insertion, activated platelets spread on the internal surface of the prosthesis and 24 hrs later numerous platelet aggregates supported by a fibrin network could be observed. However, no evidence for platelet activation could be concomitantly found in peripheral blood. A sustained increased in PGI2 formation was detected together with a progressive increase in plasma fibrinolytic activity during thrombus growth. The levels of fibrinogen as well as antithrombin III (ATIII) and heparin cofactor II (HCII) activities were steadily increased in loop-bearing animals. In conclusion, the dynamic phases of thrombus formation, in an aortic prosthesis, produce changes in vascular function and in hemostatic factors at the level of systemic blood. PMID- 8731512 TI - Use of an active center-directed plasmin inhibitor elucidates the multiplicity of plasmin actions. AB - In our studies, designed to synthesize an active center-directed plasmin (PL) inhibitor, N-(4-aminomethylbenzoyl)-4-(3-picolyloxy)-L-phenylalanine n-hexylamide dihydrochloride (PASI-535) was found. We characterized PASI-535 and analyzed the actions of PL, comparing with those of PASI-535 and tranexamic acid (t-AMCHA). (1) PASI-535 strongly inhibited not only fibrinolysis (IC50: 2.9 x 10(-6) M) but also amidolysis (Ki value: 2.9 x 10(-6) M) and fibrinogenolysis (IC50: 4.5 x 10( 6) M) induced by PL. While t-AMCHA which strongly inhibited fibrinolysis (IC50: 6.0 x 10(-5) M), rarely inhibited amidolysis (Ki value: 4.0 x 10(-2) M) and fibrinogenolysis (IC50: 1.0 x 10(-2) M). (2) PL is able to liberate kinins by degrading kininogen. This kinin-generation by PL was inhibited by 2 x 10(-5) M PASI-535. However, it was little inhibited even by 1 x 10(-3) M t-AMCHA. (3) The inhibitory effect of PASI-535 and t-AMCHA on tumor growth was studied. In sarcoma 180 bearing mice, ascites retention and the increase of tumor cells were markedly suppressed by subcutaneous injection of PASI-535, either 30 mg/kg/day or 50 mg/kg/day, for 5 days, and the inhibitory effect was dose-dependent. Although t AMCHA also reduced both ascites retention and the increase of tumor cells, it needed approximately 40 times (2 g/kg/day) the amount of PASI-535 to exert these effects. PASI-535 may be a useful tool in analyzing the multiplicity of PL actions. Moreover, PASI-535 can be used as an antifibrinolytic drug which has a mechanism of function different from that of t-AMCHA. PMID- 8731513 TI - Hemostasis activation during sclerotherapy of lower extremity varices. AB - The influence of compression sclerotherapy upon hemostasis activation was investigated in 41 consecutive patients with lower extremity varices by serial measurement of thrombin-antithrombin III complexes (TAT), D-dimer, fibrinogen and C-reactive protein (CRP). Blood sampling was carried out before operation and on the 7th and 28th post-operative day in patients randomly assigned to either the control group (n = 18), in which high ligation of sapheno-femoral junction and local excision of varices were performed, or the sclerotherapy group (n = 23) in which the comparable surgical intervention and compression sclerotherapy using hypertonic saline were performed simultaneously. In both groups, the TAT, D-dimer and fibrinogen concentrations at day 7 were significantly elevated compared to the value before operation while CRP showed no significant change during the observation period. In the sclerotherapy group, higher incidence of superficial thrombosis was observed and the TAT concentration at day 7 was significantly higher than that in the control group (p < 0.01), and the TAT at day 28 was still significantly elevated compared to the pre-operative level (p < 0.05). However, no relationship between TAT and D-dimer concentrations and the extent of superficial thrombosis was observed. We conclude that compression sclerotherapy for lower extremity varices causes latent activation of coagulation system and can be a risk factor for venous thromboembolism. PMID- 8731514 TI - Pulmonary vascular injury induced by hemorrhagic shock is mediated by P-selectin in rats. AB - To investigate whether the P-selectin-mediated leukocyte adhesion to the endothelial cells is involved in pulmonary vascular injury after hemorrhagic shock, we examined the effect of an anti-P-selectin monoclonal antibody (MAb PB1.3) on the pulmonary accumulation of leukocytes and the subsequent pulmonary vascular injury observed after hemorrhagic shock in rats. Two hours after hemorrhagic shock, pulmonary accumulation of leukocytes, as evaluated by measuring myeloperoxidase activity, began to increase and peaked after 6 hours. Pulmonary vascular injury, as evaluated by the extravascular leakage of 125I albumin, was significantly increased 6 hours after hemorrhagic shock. MAb PB1.3 significantly prevented both the pulmonary accumulation of leukocytes and subsequent pulmonary vascular injury. MAb PNB1.6, an anti-P-selectin monoclonal antibody incapable of inhibiting P-selectin-mediated leukocyte adhesion, did not prevent either of these effects. These observations strongly suggest that the pulmonary sequestration of leukocytes and the subsequent pulmonary vascular injury after hemorrhagic shock are mediated by P-selectin. PMID- 8731515 TI - Reduced beta-adrenergic receptor-coupled Gs protein function and Gs alpha immunoreactivity in mononuclear leukocytes of patients with depression. AB - beta-Adrenergic receptor-coupled Gs protein function was measured in 26 depressed patients through cholera toxin-sensitive, isoproterenol-induced increases in 3H Gpp(NH)p binding capacity to mononuclear leukocytes (MNL). Highly significant reductions in receptor-coupled Gs protein function were observed in the depressed patients: 2.0 +/- 1.3% increases in guanine nucleotide-binding capacity, in comparison with the control group values of 28.3 +/- 6.9%. Similar reductions in Gs protein function were detected in both uni- and bipolar depressed patients. A significant negative correlation was found between receptor-coupled Gs protein measures and the severity of depression. Adding semiquantitative measures of MNL Gs alpha through immunoblot analysis by use of polyclonal antibodies against Gs alpha subunit, it was found that Gs alpha relative immunoreactivity was reduced from 100 +/- 2.0% in the control group of subjects to 75.9 +/- 2.3% in the depressed patients. We have previously described hyperfunctional Gs proteins in leukocytes of patients with mania. The present findings of reduced function of Gs in depressed patients suggests receptor-coupled Gs protein activity as a biochemical parameter indicatory of the affective state. Reduced receptor-coupled Gs protein function may reflect reduced levels of the beta-adrenergic receptor previously shown in leukocytes of depressed patients; however, our complementary immunoblot studies suggest a direct, postreceptor, quantitative, and functional reduction in Gs protein in MNL of depressed patients. PMID- 8731516 TI - D,L-fenfluramine challenge test: experience in nonpatient sample. AB - The plasma prolactin response to a single-dose fenfluramine challenge is increasingly utilized in psychiatric research as an indirect and noninvasive measure of central serotonergic activity. However, the influences of age, gender, and body weight on prolactin response and characterization of physical and psychological symptoms evoked by fenfluramine remain poorly studied. In the current study, 83 nonpatient male and female volunteers, 25-60 years old, were administered a standardized fenfluramine challenge test (60 mg). Serial blood samples for plasma drug concentration and plasma prolactin concentration were obtained and side effects reported by participants were recorded. Analyses revealed that both plasma drug concentration and prolactin response were correlated with weight-relative dose (r = 0.43 and r = 0.38, respectively; p < 0.001). No significant relationship was noted between prolactin response and either age or gender. Symptoms during fenfluramine challenge were reported by 90% of subjects, most commonly fatigue, headache, lightheadedness, and difficulty concentrating. Overall side effect severity was related to weight-relative dose (r = 0.26; p < 0.05) and prolactin response (r = 0.42; p < 0.001). We conclude that fenfluramine challenge results should be reported as change in plasma prolactin relative to dose, and that in nonpatient samples the test is associated with frequent side effects. PMID- 8731517 TI - Corpus callosum morphology, as measured with MRI, in dyslexic men. AB - To test the hypothesis of anomalous anatomy in posterior brain regions associated with language and reading, the corpus callosum was imaged in the midsagittal plane with magnetic resonance. The areas of the anterior, middle, and posterior segments were measured in 21 dyslexic men (mean age 27 yrs, SD 6) and in 19 matched controls. As predicted, the area of the posterior third of the corpus callosum, roughly equivalent to the isthmus and splenium, was larger in dyslexic men than in controls. No differences were seen in the anterior or middle corpus callosum. The increased area of the posterior corpus callosum may reflect anatomical variation associated with deficient lateralization of function in posterior language regions of the cortex and their right-sided homologues, hypothesized to differ in patients with dyslexia. PMID- 8731518 TI - Elevated cerebrospinal fluid corticotropin-releasing factor in Tourette's syndrome: comparison to obsessive compulsive disorder and normal controls. AB - Stress- and anxiety-related fluctuations in tic severity are cardinal features of Tourette's syndrome (TS), and there is evidence for involvement of noradrenergic mechanisms in the pathophysiology and treatment of the disorder. To examine further the pathobiology of this enhanced vulnerability to stress and anxiety, we measured central activity of corticotropin-releasing factor (CRF) in patients with TS and the related condition, obsessive compulsive disorder (OCD). Lumbar cerebrospinal fluid (CSF) was obtained in a standardized fashion for measurement of CRF from 21 medication-free outpatients with TS, 20 with OCD, and 29 healthy controls. The TS patients had significantly higher levels of CSF CRF than both the normal controls and the OCD patients. However, there was no difference in CSF CRF between the OCD patients and the normal controls. Group differences in CSF CRF were unrelated to current clinical ratings of depression, anxiety, tics, and obsessive compulsive behaviors. Although the functional significance of this finding remains to be elucidated, these results are consistent with the hypothesis that stress-related neurobiological mechanisms may play a role in the pathobiology of TS. PMID- 8731519 TI - Differential tolerance to biological and subjective effects of four closely spaced doses of N,N-dimethyltryptamine in humans. AB - Tolerance of the behavioral effects of the short-acting, endogenous hallucinogen, N,N-dimethyltryptamine (DMT) is seen inconsistently in animals, and has not been produced in humans. The nature and time course of responses to repetitive, closely spaced administrations of an hallucinogenic dose of DMT were characterized. Thirteen experienced hallucinogen users received intravenous 0.3 mg/kg DMT fumarate, or saline placebo, four times, at 30 min intervals, on 2 separate days, in a randomized, double-blind, design. Tolerance to "psychedelic" subjective effects did not occur according to either clinical interview or Hallucinogen Rating Scale scores. Adrenocorticotropic hormone (ACTH), prolactin, cortisol, and heart rate responses decreased with repeated DMT administration, although blood pressure did not. These data demonstrate the unique properties of DMT relative to other hallucinogens and underscore the differential regulation of the multiple processes mediating the effects of DMT. PMID- 8731520 TI - Effect of tetrahydroaminoacridine on sleep in healthy subjects. AB - The present study investigated the impact of the cholinesterase inhibitor tetrahydroaminoacridine (THA; tacrine) on sleep in healthy subjects. According to the reciprocal interaction model of non-rapid eye movement (NREM) and REM sleep regulation, which postulates a primary role in cholinergic neurotransmission for the initiation and maintenance of REM sleep, it was expected that THA would lead to an earlier onset of REM sleep. In 12 healthy subjects aged from 21 to 50 years two different doses (20 mg, 40 mg) were administered 1 hour prior to bed time and compared to placebo. Only the higher dose of THA significantly shortened REM latency. No other significant effects on sleep architecture were observed, although administration of 40 mg tacrine was associated with a decrease in sleep efficiency and a prolongation of sleep latency. Blood plasma levels of tacrine and its metabolite 1-hydroxytacrine measured prior to sleep and during the first 90 min of sleep were significantly correlated with the onset of REM sleep in relation to the timing of drug administration (only for the 20 mg dose). The reversible cholinesterase inhibitor THA exerts effects on REM latency comparable to those observed with other cholinomimetic agents. PMID- 8731522 TI - Enhanced retrieval of unpleasant memories influenced by shock controllability, shock sequence, and rat strain. AB - The propensity to recall unpleasant events may be related to depression and posttraumatic stress disorder. This study examined the extent to which the recall of a previously unpleasant event (i.e., passive avoidance training) may be influenced by another aversive event. The other aversive event was tail shock. Since the Wistar Kyoto (WKY) rat strain has been proposed as an animal model of depressive behavior, this study was conducted with WKY and Wistar rats. Parameters manipulated included shock controllability, shock sequence (i.e., tail shock before avoidance training versus tail shock after training), and rat strain. Performance of the passive avoidance (PA) response was greater in WKY rats. Exposure to inescapable tail shock was related to greater PA performance compared to exposure to escapable or no-shock treatments. Tail shock prior to PA training led to a greater recall of the PA response. The magnitude of the PA response was influenced by the rat strain, shock controllability, and shock sequence. The applicability of these data to the memory bias phenomenon in depression is discussed. PMID- 8731521 TI - Depression in multiple system atrophy and in idiopathic Parkinson's disease: a pilot comparative study. AB - Multiple system atrophy (MSA) is a disease causing parkinsonism in which response to levodopa is classically absent, poor, or transient. Idiopathic Parkinson's disease (IPD) itself, which responds favorably to levodopa, has been associated with the development of disease-related depression. Over and above the clinical and pathological characteristics of IPD, MSA causes additional, more widespread, clinical and pathological deficits. We have compared motor disability and mood in 12 patients with MSA and 12 with IPD. There was more severe motor disability, but no clinical evidence of depression among the MSA patients studied, and their Beck Depression Inventory scores did not differ significantly from the group with IPD. We conclude that depression does not appear to be more common in MSA than in IPD. PMID- 8731523 TI - Immediate recurrence of psychotic symptoms after discontinuation of clozapine: a case report. PMID- 8731524 TI - Recent cocaine use decreases negative signs in acute schizophrenia: a case study over two consecutive admissions. PMID- 8731525 TI - Adverse symptomatology and suicide associated with the use of methylenedioxymethamphetamine (MDMA; "Ecstasy"). PMID- 8731526 TI - Do semantic priming effects correlate with sensory gating in schizophrenia? PMID- 8731527 TI - A brain single photon-emission computed tomography (SPECT) study of generalized social phobia. PMID- 8731528 TI - A comparison of the ability of myo-inositol and epi-inositol to attenuate lithium pilocarpine seizures in rats. PMID- 8731529 TI - Advance directives. PMID- 8731530 TI - Ethics without abstraction: squaring the circle. PMID- 8731531 TI - Non-therapeutic (elective) ventilation of potential organ donors: the ethical basis for changing the law. AB - Non-therapeutic ventilation of potential organ donors would increase the supply of kidneys for transplantation. There are no major ethical objections to it. The means of permitting it are forbidden by laws with an ethical basis. A law permitting it would need an ethical basis. Introducing a third legal method of diagnosing death would be unethical. Expanding the power of the advance directive to permit procedures involving minimal harm would be ethical but not helpful. Extending the power of proxies to permit specific non-therapeutic procedures which caused or risked minimal harm to incompetent patients is the best way forward. PMID- 8731533 TI - Bioethics and caring. AB - The author agrees with the critiques of moral theory offered by such writers as Bernard Williams and Alasdair MacIntyre, and uses ideas from Heidegger and Levinas to argue that caring is an ontological structure of human existence which takes two forms: caring about on self (which he calls our "self-project") and caring-about-others. This dual form of caring is expressed on four Aristotelian levels of human living which the author describes and illustrates with reference to the phenomenon of pain. It is concluded from this analysis that traditional notions of morality as imposing obligations should give way to an understanding of ethics as the social forms given to our caring for ourselves and for others. A number of implications for ethical theory are sketched out with the conclusion that virtue theory should be preferred and that the model could be worked out more fully to show that virtue theory can be internalist, particularist, pluralist, personalist and objectivist. PMID- 8731532 TI - Research ethics committee audit: differences between committees. AB - The same research proposal was submitted to 24 district health authority (DHA) research ethics committees in different parts of the country. The objective was to obtain permission for a multi-centre research project. The study of neonatal care in different types of unit (regional, subregional and district), required that four health authorities were approached in each of six widely separated health regions in England. Data were collected and compared concerning aspects of processing, including application forms, information required, timing and decision-making. The key finding was that ethics committees received and processed the applications variably, reflecting individual factors and local problems. To improve consensus and facilitate multicentre studies, standard forms and instructions are suggested and the establishment of a national committee or advisory group advocated. PMID- 8731534 TI - What are the limits to the obligations of the nurse? AB - This paper enquires into the nature and the extent of the obligations of nurses. It is argued that nurses appear to be obliged to undertake supererogatory acts if they take clause one of the United Kingdom Central Council for Nursing, Midwifery and Health Visiting (UKCC) Code of Professional Conduct seriously (as, indeed, they are required to do). In the first part of the paper, the nature of nursing obligations is outlined, and then the groups and individuals to whom nurses have obligations are identified. Following a brief discussion of the moral foundation of the nurse's obligations to her/his employer, a common conflict of obligations is identified. Then a distinction is drawn between ordinary and extraordinary moral standards. Appreciation of this is necessary for an understanding of the criterion of what constitutes a supererogatory act. By the definition of supererogatory acts proposed below, it is suggested that actions such as whistleblowing satisfy that definition. PMID- 8731535 TI - Betting your life: an argument against certain advance directives. AB - In the last decade the use of advance directives or living wills has become increasingly common. This paper is concerned with those advance directives in which the user opts for withdrawal of active treatment in a future situation where he or she is incompetent to consent to conservative management but where that incompetence is potentially reversible. This type of directive assumes that the individual is able accurately to determine the type of treatment he or she would have adopted had he or she been competent in this future scenario. The paper argues that this assumption is flawed and provides theoretical and empirical evidence for this. If the assumption is false, and those taking out advance directives do not realise this, then the ethical bases for the use of these advance directives-the maximisation of the individual's autonomy and minimisation of harm-are undermined. The paper concludes that this form of advance directive should be abolished. PMID- 8731536 TI - Limiting risks by curtailing rights: a response to Dr Ryan. AB - It has been argued that the inherent risks of advance directives made by healthy people are disproportionate to the potential benefits, particularly if the directive is implementable in cases of reversible mental incapacity. This paper maintains that the evidence for such a position is lacking. Furthermore, respect for the principle of autonomy requires that individuals be permitted to make risky choices about their own lives as long as these do not impinge on others. Even though health professionals have an obligation to try and ensure that patients have appropriate information about possible future treatment options, they cannot predict and describe every eventuality but nor can they disregard firm decisions knowingly made on the basis of incomplete information by competent adults. To attempt to do so would be to reinstate notions of medical paternalism which are contrary to current public expectations. PMID- 8731537 TI - Genetic testing and early diagnosis and intervention: boon or burden? AB - The possibility of early diagnosis and intervention is radically changed by the advent of genetic testing. The recent report of the Nuffield Council on Bioethics is timely and helpful. I have suggested, that not only the severity of the disability indicated by genetic information, and the accuracy of the data, ought to govern the approach to the implementation of screening for genetic disorders. In addition, assessment of the value of the information to those involved should be considered. The efficacy of the available therapeutic measures, combined with the prognostic data are important indices of the value of the information. These measures fall into three categories and thus indicate that three different courses of intervention may be appropriate. Three approaches to diagnosis and intervention are then outlined, drawing on the experience of various clinical initiatives. PMID- 8731538 TI - Ethical considerations of the perinatal necropsy. AB - The perinatal necropsy is an important investigation following fetal or neonatal loss. Legal requirements on registration decree that consent is needed before necropsy can proceed in some of these babies. However, there are ill-defined grey areas which are open to legal and ethical difficulties. This paper discusses the problems that can arise with consent for a necropsy in the perinatal period. Some of these problems are clearly legal or ethical but all can cause distress to parents at a time of grief and bereavement. The issues may not be readily resolved but public debate and ad hominem decisions on each perinatal loss may help to alleviate the problems. PMID- 8731539 TI - An obligation to provide abortion services: what happens when physicians refuse? AB - Access to abortion services in the United States continues to decline. It does so not because of significant changes in legislation or court rulings but because fewer and fewer physicians wish to perform abortions and because most states now have "conscientious objection" legislation that makes it easy for physicians to refuse to do so. We argue in this paper that physicians have an obligation to perform all socially sanctioned medical services, including abortions, and thus that the burden of justification lies upon those who wish to be excused from that obligation. That is, such persons should have to show how requiring them to perform abortions would represent a serious threat to their fundamental moral or religious beliefs. We use current California law as an example of legislation that does not take physicians' obligations into account and thus allows them too easily to declare conscientious objection. PMID- 8731540 TI - A century of controversy surrounding posttraumatic stress stress-spectrum syndromes: the impact on DSM-III and DSM-IV. AB - The authors describe historical clinical reports that preceded the development of criteria for posttraumatic stress disorder (PTSD) and influenced the formation of PTSD in DSM-IV. These reports were identified from extensive search of 19th- and 20th-century American and European medical literature. Relevant findings from the most representative reports are described and discussed. Since the mid-19th century, clinical syndromes resembling PTSD have been described. However, understanding of PTSD has been complicated by questions of nomenclature, etiology, and compensation. Nomenclature placed PTSD syndromes under existing psychiatric disorders: traumatic hysteria, traumatic neurasthenia, or traumatic neurosis. Etiological issues have been concerned often solely with organic factors, pre-existing personality impairments, intrapsychiatric conflicts, and social factors. Only after World War II and the concentration camp experiences did the role of severe trauma in PTSD become recognized. Even though controversy remains, much progress in understanding PTSD has been made. PMID- 8731541 TI - Survivors of torture and organized violence: demography and diagnosis. AB - Sixty patients, with a history of torture or other repressive state violence, newly referred to two psychiatrists, were assessed using a standard instrument. Thirty one met the diagnostic criteria for posttraumatic stress disorder (PTSD) and 21 met the criteria for MDD. Relationships between these diagnoses and history of trauma, loss of health, and social losses were investigated. Sexual torture is associated with an avoidance reaction. PMID- 8731542 TI - The long-term sequelae of sexual abuse: support for a complex posttraumatic stress disorder. AB - This study examined the relationship between childhood sexual abuse and symptoms of a newly proposed complex posttraumatic stress disorder or disorder of extreme stress not otherwise specified (DESNOS). Compared to 34 women without histories of sexual abuse, 74 survivors of sexual abuse showed increased severity on DESNOS symptoms of somatization, dissociation, hostility, anxiety, alexithymia, social dysfunction, maladaptive schemas, self-destruction and adult victimization. In addition, a logistic regression found that a complex of symptoms representing DESNOS was significantly related to a history of sexual abuse. Consistent with other studies, the results of this study provide support for the idea that symptoms of DESNOS characterize survivors of sexual abuse. PMID- 8731543 TI - Psychological adaptation of anxiety disorder patients following repeated exposure to emergency situation. AB - Thirty one patients in treatment for anxiety disorders and 31 controls were interviewed within hours after both the first and second Iraqi missile attacks on Israel during the Gulf war. After the first attack patients did not report higher anxiety levels, nor were they more pessimistic about the war and their fate in the war than the control subjects. Anxiety disorder patients tended to be engaged in cognitive-behavioral tactics for self-calming, while control subjects clearly preferred to cope by interacting with their social and physical environments. Following the second missile bombardment, patients were more inclined to retain their initial levels of anxiety and pessimism, while controls seem to have better adapted and showed significant improvements in those variables. The results are discussed in terms of coping skills and vulnerability as factors influencing adaptation to prolonged emergency situations. PMID- 8731544 TI - Initial posttraumatic stress responses following motor vehicle accidents. AB - Posttraumatic stress was assessed in 114 motor vehicle accident (MVA) victims within 2 weeks of hospital admission. Approximately one third of patients reported high levels of posttraumatic stress anxiety. Intrusion symptoms were best predicted by fear of the MVA and absence of head injury. Avoidance symptoms were best predicted by fear of the accident and recent stressful events. Findings indicate that intrusion and avoidance responses to trauma are mediated by different patterns of injury-related and response-related factors. PMID- 8731545 TI - Social phobia and PTSD in Vietnam veterans. AB - Posttraumatic stress disorder (PTSD) is the most prevalent psychological disorder experienced by Vietnam veterans. However, there are many other disorders and problems of adjustment, like social anxiety and social phobia, that have not been fully investigated in this population. This study examined the prevalence of social phobia and the comorbidity of social phobia and PTSD, and tested out a theory of the etiology of social anxiety in trauma victims. Forty one Vietnam combat veterans were interviewed and completed self-report measures assessing PTSD and social phobia. Adversity of homecoming was also assessed. Using a conservative multi-method assessment approach, 32% of the sample were found to be positive for both social phobia and PTSD. Veterans with PTSD were significantly more likely to carry an additional diagnosis of social phobia as compared to veterans without PTSD. Adversity of homecoming and shame about one's experience in Vietnam were significant predictors of current level of social anxiety over and above the effects of pre-military anxiety and severity of combat exposure. These observations suggest that social anxiety and social phobia may be significant problems among individuals with PTSD. Further, these findings offer preliminary support for the theory that posttrauma environment may impact upon the later development of social anxiety. PMID- 8731546 TI - Dissociative experiences and posttraumatic stress disorder among female victims of criminal assault and rape. AB - This study examines the relationship between posttraumatic stress disorder (PTSD) and dissociative experiences in a sample of 158 recent female assault victims (74 rape, 84 nonsexual assault) and 46 comparison subjects who had not been assaulted within the last year. Results indicated that victims had elevated scores on Dissociative Experiences Scale (DES) as compared to the comparison subjects, but that this elevation was not as high as for other traumatized samples. The level of dissociation reported by assault victims declined significantly over the three month course of the study. DES scores were related to PTSD diagnosis and symptom severity, but only among nonsexual assault victims. In rape victims, there was no correlation between dissociation and PTSD. Recent victims with a history of childhood sexual abuse were significantly more dissociative than those who did not report such a history. These results are discussed with regard to vulnerability factors for developing PTSD subsequent to a criminal assault. PMID- 8731547 TI - Effectiveness of short-term specialized inpatient treatment for war-related posttraumatic stress disorder: a role for adventure-based counseling and psychodrama. AB - Psychological tests were administered to 24 participants of an inpatient posttraumatic stress disorder (PTSD) treatment program both immediately before and following completion of treatment. Responses were compared to a treatment/wait list comparison group composed of 24 subjects awaiting entry into the program. All treatment/wait list comparison group subjects received weekly PTSD outpatient group therapy. Significant improvements were found in the inpatient treatment group in areas of hopelessness, feelings of guilt and shame, loneliness, and emotional expressiveness. Other indices of psychological functional, including interpersonal skills, gender role stress, anxiety, anger, and PTSD symptomatology did not change significantly in response to treatment. No positive changes in any area of psychological function occurred in the treatment/wait list comparison group. Implications for PTSD and areas of future research are discussed. PMID- 8731548 TI - The revised Civilian Mississippi Scale for PTSD: reliability, validity, and cross language stability. AB - Examined in two studies the psychometric properties of a revised 30-item version of the civilian form of the Mississippi Scale for Combat-Related Posttraumatic Stress Disorder (PTSD) developed by Keane, Caddell, and Taylor (1988). Study 1, whose sample was composed of 37 bilingual adults who had experienced a variety of traumatic events, was undertaken primarily to examine the linguistic equivalence of a Spanish translation of the scale. High cross-language stability was demonstrated, and both English and Spanish versions showed high internal consistency. Study 2, which used a sample of 404 victims of Hurricane Andrew, provided additional evidence of scale reliability and also showed that the scale correlates in meaningful ways with known traumatic stressors. Together the results indicate that the scale is applicable to different populations and events and constitutes a valid and reliable self-report, measure of PTSD. PMID- 8731549 TI - Modes of long-term coping with trauma memories: relative use and associations with personality among Vietnam veterans with chronic PTSD. AB - Little is known about how individuals who develop chronic posttraumatic stress disorder (PTSD) cope with recurring trauma memories, or how enduring personality characteristics influence such coping. Focusing on 110 hospitalized Vietnam combat veterans with chronic PTSD, this exploratory study assessed the relative frequency of using eight ways of coping with war memories, and associations between relative use of these strategies and eight dysfunctional personality styles. As a secondary issue, associations between coping strategies, combat exposure, and PTSD severity were also examined. Consistent with prior findings, these veterans predominantly used emotion-focused and avoidant strategies to cope with war memories. Differing personality styles and relative use or nonuse of particular coping strategies were also associated in psychologically coherent ways. These preliminary findings are discussed in relation to methodologic and future research issues. PMID- 8731550 TI - Appraisal of control and coping with air-raids. AB - This study investigated the appraisal of control (secondary appraisal) and coping as reactions to the air-raids in the city of Zagreb. Coping was assessed with the Croatian version of Ways of Coping (Lazarus & Folkman, 1984) and secondary appraisal was operationalized as control over the occurrence and over the outcome/consequences of the air-raid. The hypotheses that appraisal of low control would yield more emotion-focused and passive coping, and that appraisal of high control would be linked with problem-focused and active coping, were not confirmed. The results were interpreted in terms of so-called emotional habituation. Additionally, some unique characteristics of the air-raid were identified and explained as the key variables in the interpretation of the obtained results: an air-raid is a very specific stressful situation, it is homogeneous in content, and it is a chronic and repeated stressor. The problems with generalization of these data to all stressful events were pointed out. PMID- 8731551 TI - Interpersonal and self-reported hostility among combat veterans with and without posttraumatic stress disorder. AB - The present study investigated self-reported and interpersonal hostility in 70 Vietnam combat veterans with and without posttraumatic stress disorder (PTSD) and 60 comparison community volunteer subjects. Veterans were 50 help-seeking, male Vietnam combat veterans with PTSD and 20 non-help-seeking male combat veterans without PTSD. Vietnam veterans with PTSD not only reported more hostility than non-PTSD veterans and healthy community volunteers, but also reacted behaviorally with more hostility during an interpersonal interaction. Compared to veterans without PTSD, veterans with PTSD reported significantly higher levels of hostility and demonstrated significantly greater non-verbal expressions of hostility during an interpersonal task. These results suggest that the level of hostility in PTSD combat veterans may be high as compared to comparison groups. The implications of these results and possible research directions are presented. PMID- 8731552 TI - Race and outcome of treatment for veterans suffering from PTSD. AB - This study examines the relationship between racial group membership and psychometrically measured outcomes 4, 8 and 12 months after program entry in a program for veterans seeking treatment for war-related posttraumatic stress disorder (PTSD). Longitudinal assessment data were gathered and used to compare service use, clinicians' improvement ratings and psychometrically assessed clinical change among Black (n = 122) and White (n = 403) veterans treated at six geographically diverse sites. There were no significant differences between Blacks and Whites on any of the clinicians' improvement ratings, or on 13 of the 17 outcome measures. In this prospective study of veterans suffering from long standing, severe PTSD, no consistent or sustained differences were observed between racial groups, in improvement whether measured as psychometric change or by clinicians' ratings. PMID- 8731553 TI - Examining comorbidity and posttraumatic stress disorder in a Vietnam veteran population using the MMPI-2. AB - Examined the discriminant validity of the MMPI-2 in assessing comorbidity in a posttraumatic stress disorder (PTSD) Vietnam veteran population. The Structured Clinical Interview for the DSM-III-R (SCID) was used to diagnose veterans and to classify them into four groups: PTSD Only, PTSD with mood disorders, PTSD with other anxiety disorders, and PTSD with mood and anxiety disorders. All groups had clinical elevations on scales F, 1, 2, 3, 4, 6, 7, 8, 0, PK, and PS, with peak elevations on scales 8, 7, and 2. The PTSD Only group's MMPI-2 scores were not significantly lower than other groups' scores. The PTSD+Mood/Anxiety group was significantly more elevated on scales 2 and 7 than the PTSD Only and PTSD+Anxiety group but did not otherwise show significantly higher scale elevations than others groups. No significant differences existed between groups on scales F, L, K, PK, and PS. Implications of these results for PTSD and the current diagnostic system are explored. PMID- 8731554 TI - Childhood factors and war zone stress in chronic PTSD. AB - This study assessed the predictive validity of combat factors and selected premilitary variables (i.e., childhood physical abuse, substance abuse in the family of origin, or being raised in a nonadaptive or noncohesive family) on posttraumatic stress disorder (PTSD) group membership. In addition, it assessed the correlation of combat exposure and selected premilitary variables with the severity of PTSD symptomology. Ninety-three male Vietnam combat veterans with PTSD were compared to 82 male Vietnam combat veterans without the disorder. The results of two hierarchical logit analyses identified combat exposure as the best predictor of PTSD group membership. However, physical punishment was also found to significantly predict group membership when entered first in the analyses. Furthermore, multiple regression analyses conducted with the PTSD group alone found that both combat exposure and physical abuse predicted greater PTSD symptomology. These findings suggest that childhood physical abuse as well as military trauma should be addressed in the assessment and treatment of chronic PTSD patients. PMID- 8731555 TI - Antinuclear antibodies and thyroid function in sexually abused girls. AB - Sexually abused girls manifest dysregulation of physiological stress response systems. In this exploratory investigation, 14 sexually abused and 13 control girls, ages 8-15 years, recruited from a prospective, longitudinal study, underwent plasma antinuclear antibody and thyroid function tests. Thyroid function tests and plasma antinuclear antibody titers did not differ between sexually abused and control girls. However, a significantly higher incidence of plasma antinuclear antibody titers was seen in abused subjects when compared with the frequency of positive antinuclear antibody titers in a sample of 22 adult healthy female volunteers, ages 20-58 years. These findings suggest that sexually abused girls may show evidence of an alteration in normal immune homeostatic function. PMID- 8731556 TI - The impact of cohesive groups in the trauma recovery context: police spouse survivors and duty-related death. AB - This paper examines the impact of surviving spouse social interactions on psychological distress following the death of a police officer. It was hypothesized that satisfactory interactions within the police work group would lower distress, and unsatisfactory interactions outside the police environment (justice system, media, community persons) would increase distress. Results indicated that increased quality of interaction with police groups lowered psychological distress scores. Despite increased satisfaction with groups outside of policing, spouse's distress still increased, suggesting that satisfaction with and support by police groups appears to ameliorate distress more effectively than others. These findings suggest that police agencies and personnel may be helpful to surviving spouses after the death of an officer and should formulate policy to provide long term contact and assistance. PMID- 8731557 TI - Program evaluation of an intervention approach for staff assaulted by patients: preliminary inquiry. AB - Crisis intervention procedures for emergency services personnel and healthcare providers are increasingly being utilized to address the psychological sequelae of acute stress during critical incidents. While anecdotal evidence suggests the efficacy of these approaches, more formal experimental inquiry has been limited. This brief report outlines one strategy for conducting program evaluation of a crisis intervention approach for staff who are assaulted by patients. This outcome design was implemented in multiple sites, even as the service was being delivered. PMID- 8731558 TI - The influence of hunting pressure on the social behavior of vertebrates. AB - The study of the influence of human hunting pressure on the social behavior of vertebrates requires a large background on both the hunting pressure and the social behavior of the species. Literature about this subject is relatively scarce. Most of the papers are restricted to shifts in demography, and are generally "species-specific". However, human hunting pressure can not only affect demography but also some factors of the social behavior of a species such as parental care, territoriality, reproductive behavior, group-size, fraying behavior, mating system, and intraspecific competition. The presence of hunters in the area can lead the animals to move out of their original home-rangers. This can breakup the social structure and can cause long-term effects on the demography of hunted as well as nearby populations, amplifying the impact of hunting. On the other hand, harvest can be compensated in territorial species by "surplus population" of non-territorial individuals. Moreover, alternatively, an increase in the reproductive activity, as a response to hunting pressure, can compensate its effects in some species. Understanding the implications of hunting pressure in the social behavior of vertebrates might be decisive to the establishment of wildlife management and conservation programs. Manipulative experiments might help us to model such complex interactions. The occurrence of qualitative and quantitative changes in vertebrate social behavior directly or indirectly related to human hunting pressure is reviewed and analyzed in this paper. PMID- 8731559 TI - Targeting the vasculature of solid tumours. PMID- 8731560 TI - The genetic basis for mental retardation. PMID- 8731561 TI - Association of HLA types A1-B8-DR3 and B27 with rapid and slow progression of HIV disease. AB - We examined how HLA types A1-B8-DR3 and B27 were related to progression of clinical disease and rate of loss of CD4 lymphocytes in the Edinburgh City Hospital cohort of HIV-positive patients, mainly injection drug users. Patients (n = 692) were prospectively followed from 1985 through March 1994. Accurately estimated seroconversion times were determined retrospectively for a subgroup of 313 (45%). Of 262 patients (39%) who were fully or partially HLA typed, 155 (50%) had known seroconversions. Of 34 patients typed positive for A1-B8-DR3, 29 progressed to CDC stage IV, 22 to AIDS and 20 died. Twelve patients were typed positive for B27; six of these progressed to CDC stage IV, one to AIDS and none died. In a proportional hazards analysis of the 313 patients with known seroconversions, A1-B8-DR3 was significantly associated with covariate-adjusted relative risks of 3.7 (95% CI 1.9-7.2), 3.1 (1.6-6.0) and 1.9 (1.1-3.2) for progression from seroconversion to death, AIDS and CDC stage IV, respectively. Events for B27 were too rare to include B27 in analyses to death and AIDS, but B27 was significantly associated with slower progression to CDC stage IV (0.3, CI 0.1-0.9). Random effects growth curve models were used to estimate individual rates of loss of square root CD4 count and loss of CD4 percentage, for 603 and 617 patients, respectively. A1-B8-DR3 was associated with rapid loss of both markers (p = 0.02 and p = 0.01, respectively); B27 was associated with slow loss of both markers (p = 0.04 and p < 0.005). PMID- 8731562 TI - Serum oligoclonal IgG is a common and persistent finding in multiple sclerosis, and has a systemic source. AB - Synthesis of oligoclonal IgG within the central nervous system is a well established feature of multiple sclerosis. The occurrence of oligoclonal IgG in the serum of patients with multiple sclerosis has received little attention. We detected such a serum response in 20/45 consecutive patients (44%, 95% CI 30-59%) but in only 3/41 age- and sex-matched healthy controls (p < 0.01). We present qualitative and quantitative evidence that this oligoclonal IgG has a systemic origin. The plasma cell clones responsible for the serum response are often also represented in the intrathecal compartment. In a further study of the clinical significance of serum oligoclonal bands, in 80 patients, their presence was associated with elevated levels of intrathecal synthesis, increasing age, later disease onset and the presence of serum autoantibodies. These findings add to the evidence that there is a systemic immune disturbance in multiple sclerosis. PMID- 8731563 TI - Adhesion molecules in untreated inflammatory arthritis: synovial expression and levels in synovial fluid and serum [corrected]. AB - Adhesion molecules play a critical role in regulating leucocyte migration at sites of inflammation. The relationship of soluble forms in serum or synovial fluid (SF) to synovial membrane expression in inflammatory arthritis is controversial. We examined soluble intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and E-selectin levels in matched serum and SF, and their relationship to expression in synovium obtained at the same time in 13 patients with previously untreated inflammatory arthritis. Serum soluble (s)ICAM-1 correlated with sedimentation rate (T = 0.45), Ritchie articular index (T = 0.47) and SF sICAM-1 (T = 0.48), and SF sICAM-1 correlated with membrane ICAM-1 expression (p < 0.02). sE-selectin and sVCAM-1 levels were unrelated to disease activity or membrane expression. Membrane E-selectin expression correlated inversely with ICAM-1 expression (T = -0.57) and serum sICAM-1 (T = -0.54). Serum sE-selectin correlated inversely with membrane ICAM-1 expression (T = -0.55). The correlations observed between ICAM-1 in serum, SF, synovium and disease activity suggest that ICAM-1 could be a useful target for immunotherapy. The inverse relationship of ICAM-1 and E-selectin suggest important differences in regulation and pathogenetic roles. PMID- 8731564 TI - Induced TNF production in vitro as a test for familial Mediterranean fever. AB - We have previously demonstrated an altered pattern of tumor necrosis factor (TNF) secretion in patients with familial Mediterranean fever (FMF). To examine whether TNF determination could assist in diagnosing FMF, we stimulated heparinized blood of 51 asymptomatic FMF patients with lipopolysaccharide (LPS) and then measured TNF production in response to inducers, compared to unstimulated blood cells and to cells from a control group of 12 matched healthy subjects. Following LPS pretreatment, which induced TNF release, FMF patients produced significantly less TNF than controls, whether production was 'spontaneous' or induced by either LPS or phytohaemagglutinin (p < or = 0.003). Such 'exhaustion' did not occur in untreated cells. We then used these results to classify a further group of 29 FMF patients and 10 matched healthy controls ('validation' group) who underwent the same studies. The test correctly identified 25/29 patients as having FMF and 7/10 controls as not having FMF; a sensitivity of 86% and a specificity of 70% (likelihood ratios 2.9 (positive test) and 0.2 (negative)). Identification of a blunted TNF response following previous stimulation by a simple assay, may help the diagnosis of FMF in asymptomatic patients, provided it is interpreted in conjunction with supportive clinical data. PMID- 8731565 TI - Non-insulin-dependent diabetes mellitus: dietary monounsaturated fatty acids and low-density lipoprotein composition and function. AB - Alterations in low-density lipoprotein (LDL) composition in diabetes affect its function with respect to control of de novo cholesterol synthesis. We examined the effect of 4 weeks of an oleic-acid-rich diet on LDL composition and function in eight Type 2 diabetic and eight non-diabetic control subjects. LDL (density 1.019-1.063 g/l) was isolated by sequential ultracentrifugation. LDL composition was measured and LDL fatty acids were determined by gas liquid chromatography. Cholesterol synthesis was measured by [14C]-acetate incorporation into the freshly isolated mononuclear leucocytes. Fasting blood glucose fell from 9.3 +/- 2.0 to 8.2 +/- 1.2 mmol/l (p < 0.05) and fasting serum insulin increased from 8.3 +/- 2.8 to 10.4 +/- 5.0 mIU/l (p > 0.05) in the diabetic patients. LDL oleic acid increased in the diabetic patients from 18.8 +/- 1.8% to 22.5 +/- 1.9% (p < 0.01) and in the non-diabetic subjects from 19.9 +/- 1.8% to 23.3 +/- 2.8% (p < 0.01). The LDL-esterified to free cholesterol ratios of 3.0 +/- 0.6 and 2.7 +/- 0.2 for the diabetic and non-diabetic patients were similar, and decreased significantly (p < 0.01) to 2.4 +/- 0.5 and 2.2 +/- 0.4, respectively. Baseline [14C]-acetate incorporation was similar in the two groups, and decreased after diet from 437 +/ 239 to 249 +/- 144 ng/g cell protein (p < 0.05) in the diabetic patients. There was a negative correlation between the LDL-esterified to free cholesterol ratio and the ratio of oleic to linoleic acid in the LDL (r = -0.39, p < 0.05) and a negative correlation between fasting blood glucose and LDL oleic acid in the diabetic patients (r = -0.51, p < 0.05). Enrichment of LDL with oleic acid appears to improve its ability to regulate endogenous cholesterol synthesis in both control and diabetic subjects. In the diabetic population, the diet had a favourable effect on glycaemic control. PMID- 8731566 TI - Antibodies against Mycobacterium paratuberculosis in Crohn's disease. AB - Until recently the investigation of serological responses to mycobacteria in patients with Crohn's disease has been hindered by the considerable degree of cross-reactivity between antigens of M. paratuberculosis, and other mycobacterial subspecies. We evaluated the serological response of Crohn's disease patients to a recently identified species-specific 18 kDa protease-resistant antigen corresponding to M. paratuberculosis bacterioferritin. The 18 kDa antigen was purified from M. paratuberculosis as previously described. Serum was obtained from 40 patients with Crohn's disease, 15 with ulcerative colitis, 25 coeliac patients, and 21 normal blood donors. Antibody levels were measured by enzyme linked immunosorbent assay (ELISA), with anti-human IgA and IgG alkaline phosphatase conjugate. Antibody titres were expressed as the dilution giving 1/3 of the plateau binding value of a standard positive serum (MT/3). Disease activity of the Crohn's disease cases was assessed using the Harvey-Bradshaw index. There was no statistically significant elevation of the mean IgG or IgA MT/3 titres of Crohn's disease patients over controls. No patients had antibody titres greater than two standard deviations above the mean control MT/3 titres, and there was no significant correlation between Crohn's disease activity and level of antibody titres. These findings make it unlikely that M. paratuberculosis is of primary pathogenic importance in Crohn's disease. PMID- 8731568 TI - Targeted screening for diabetes in community chiropody clinics. AB - Population screening for diabetes mellitus is of uncertain value. We therefore assessed the value of screening amongst community chiropody clinic attenders in Liverpool. All attenders aged between 40 and 75 years during a 3-month period were offered screening by urine glucose self-testing, 2 hours post-prandially, backed up with glucose tolerance tests (GTT) for positive respondents. Of 1058 patients screened, 11 (1.0%) reported positive results, of whom four (0.4% of total) had diabetes, and two had impaired glucose tolerance (IGT). Screening costs were 11p per person, 2.06 pounds per 'positive' person, and 34.46 pounds for each newly diagnosed patient. The screening procedure was simple and highly cost-effective, but the diagnostic returns were only moderate. This may have been because of a high rate of known diabetes amongst the chiropody clinic attenders (17.3%). In view of this, routine widespread diabetes screening in chiropody clinics cannot at present be recommended. PMID- 8731567 TI - Total radical trapping antioxidant potential (TRAP) and exercise. AB - The relationship between physical activity, physical fitness and total radical trapping antioxidant potential (TRAP) was examined in the Northern Ireland Health and Activity Survey. This was a cross-sectional population study (n = 1600) using a two-stage probability sample of the population. TRAP was calculated using the sum of the individual serum antioxidant concentrations (urate, protein thiols, ascorbate, alpha tocopherol and bilirubin) multiplied by their respective stoichiometric values. Physical fitness was determined by estimation of VO2max by extrapolation from submaximal oxygen uptake, and physical activity was recorded by computer-assisted interview. Mean serum TRAP concentrations were significantly higher in males (653 +/- 8.2 mumol/l, mean +/- SEM) compared to females (564 +/- 8.0 mumol/l) (p < 0.0001). Both male and female smokers had significantly lower TRAP values than non-smokers (males p < 0.0001, females p = 0.02). In females, there was a positive relationship of TRAP with age (p < 0.001) and body mass index (p < 0.001) but a negative relationship with physical fitness (p < 0.05). The known beneficial effects of exercise and activity do not appear to be directly mediated through increased antioxidant status. PMID- 8731569 TI - A new science of health: salutology and the evolutionary perspective. PMID- 8731570 TI - Brainstem perfusion in CFS. PMID- 8731571 TI - Diagnosis, disease and illness. PMID- 8731572 TI - [Community-acquired pneumonias. Admission criteria and complicated course indicators]. AB - To evaluate the admission criteria and to select indicators that identify patients for whom hospitalization is not necessary, we studied 75 patients with community acquired pneumonia (CAP) who were admitted to a clinical service. According to Appropriateness Evaluation Protocol (AEP) only 60% of our patients justified their hospitalization (Group A) while 40% did not (Group B). The most frequent hospitalization criteria found in Group A were tachypnea (> 30x min.) (40%), respiratory failure (38%) and encefalopathy (18%). The average age in Group A was 62 versus 47 in Group B (p < 0.001). Comorbid conditions were present in 100% of Group A and 71% had two or more while only 33% of patients in Group B had two or more (p < 0.01). During the evolution, Group A had more organ failure than B (53 vs. 17%) (p < 0.001) and a longer period of hospitalization (14 vs. 9 days) (p < 0.01). The differences between groups A and B is best visualized in the incidence of sepsis (4 vs. 0%), and mortality rates (15% vs. 0%) (p < 0.05). Using the Fine risk criteria for a complicated course, we selected 14 patients from Group B, with one or more criteria (Group C) that were compared with 16 patients without them (Group D). The presence of a poor clinical status at admission was the only difference between Group D and C (79 vs. 0%) (p < 0.001). When three or more risk factors were present the differences were significant (79 vs. 6%) (p < 0.001). We conclude that the utilization of hospitalization criteria together with the risk factors for a complicated course, specifically when two or more factors per patient are present, permit the identification of a population with CAP that needs hospitalization with 71.4% sensitivity and 100% specificity. The presence of two or less risk factors in patients without admission criteria has a highly predictable negative value (100%) and anticipates an uneventful evolution without complications. PMID- 8731573 TI - [Asthma-related mortality in the city of Rosario]. AB - Accuracy of death certification and registration have been investigated in many countries because death certificates constitute the unavoidable source for studying asthma mortality. The purposes of this study were: 1) to assess the reliability of official registration of asthma death certification and 2) to describe the percentage of asthma deaths occurring outside-hospital in Rosario during 1988. All death certificates from Rosario residents over 5 years old, in which appeared the word asthma or derivatives, were studied. The asthma mortality rates from 1981 to 1989 were obtained from the "Official annual publication". The diagnosis of asthma was written somewhere on 45 certificates; but 30 certificates were interpreted as asthma being the most appropriate diagnosis and could then be coded as the cause of death. These 30 asthma related deaths occurred at the mean age of 59.23 yrs +/- 17.23 (SD), range 12-84 yrs. Twenty-two deaths out of the total of 30 occurred in an out-hospital setting (73%). Among the 30 cases, 8 subjects (aged 63.0 yr +/- 12.38) died in hospital. There was no difference between the age, sex and the death place. Autopsy was performed in only one case of 12 years old. In other 3 cases, asthma was confirmed as the cause of death through the evaluation of case records and the confidential information collected from close acquaintances. The mean asthma mortality rate from 1981 to 88 in Rosario was 5.69 +/- 1.06/10(5), and this value was significantly higher than the death rate calculated by this study (3.46/10(5); p = 0.0005, T test for one sample). The difference probably originated in the false positive certificates often related to procedures in the General Registrar Office. In other words, there was an official overestimation of asthma deaths. This was the first description of the high percentage (73%) of asthma related deaths occurring in the out-hospital setting. Finally, even when death certificates should require a further and exhaustive assessment, asthma mortality rates in Rosario might be regarded as of great concern. PMID- 8731574 TI - [Amplification of contractile response by serotonin on noradrenaline in the human umbilical artery]. AB - A progressive increase in plasma levels of serotonin (5-HT) during pregnancy has been reported. Furthermore, an increase in the concentration of noradrenaline (NA) has been described during labor. On the other hand, in different arteries it has been observed that minimal effective concentrations of 5-HT are able to produce an increase or "amplification" of the contractile response to a second agonist. From these reports, it seemed interesting to determine the existence of a synergistic interaction between 5-HT and NA in the human umbilical artery (HUA), and the probable mechanisms involved. Therefore, strips of HUA were incubated in Krebs solution at 37 degrees C and gassed with a mixture of CO2 (5%) and O2 (95%). The contractile isometric response was assessed. After a period of two hours, complete cumulative concentration-response curves to 5-HT and to NA were obtained. The maximal contractile response to NA was approximately 20% of the maximal response to 5-HT. In another series of experiments, the response to NA was enhanced by previous treatment with minimal effective doses of 5-HT (control response 0.24 +/- 0.06 g; "amplified" response 0.53 +/- 0.06 g, P < 0.01). This "amplified" response to NA was correlated with the degree of previous contraction with 5-HT, when the last was between 3 and 30% of the maximal response. However, precontractions higher than 40% abolished the responses to NA. In other experiments, the artery strips were incubated during thirty minutes with diltiazem 1 microM before agonists addition. In this condition, a decrease in the control and amplified response was observed. Therefore, we have shown the existence of an amplifying effect of 5-HT over the response to NA in the HUA, and we discuss the possible mechanisms involved and the possible clinical relevance in preeclampsia. PMID- 8731575 TI - Cytomegalovirus-associated Menetrier's disease in adults. Demonstration by polymerase chain reaction (PCR). AB - Menetrier's disease (MD) is a rare form of hypertrophic or hyperplastic gastropathy characterized by conspicuous thickening of the gastric mucosal folds and foveolar (crypt) hyperplasia. We examined the presence of cytomegalovirus (CMV) in 2 cases of MD in adults, one associated with gastric carcinoma, using the polymerase chain reaction (PCR). None of the cases showed intranuclear inclusions consistent with CMV infection. Both revealed, besides the features of MD, a peculiar pattern of "chronic active plasmacellular gastritis". Although the samples had been stored in formalin for more than 10 years CMV-DNA was recovered with good yield from both samples. The demonstration of CMV in MD in adults may helps to explain present knowledge of the complex relationships between this virus and gastric mucosa, and strongly suggests a pathogenetic role of the virus with variable phenotypic expression along the years. PMID- 8731576 TI - [Percutaneous drainage of pyogenic hepatic abscesses]. AB - Thirty-five patients with pyogenic hepatic abscesses who underwent percutaneous drainage techniques are analyzed in a study period that comprises six years (from 1986 to 1992). Of the 35 cases, 7 (20%) presented abscesses smaller than 5 cm and were drained by puncture aspiration only. All of them were cured. One or more pig tail 8 F catheters were placed in the remaining 28 patients (80%). The cure rate was 78.5%. The average drainage period comprised 15 days. Fourteen patients (40%) presented lesser complications during either the procedure in itself or the convalescence period, bacteriemia being the most frequent one. The mortality rate was 17%. PMID- 8731577 TI - [Effect of dietary supplementation with gamma-linolenic acid on the growth of a human lung carcinoma implanted in nude mice]. AB - We have studied the effect of a gamma-linolenic acid (18:3 n-6, GLA)-supplemented diet on the growth of a human lung mucoepidermoid carcinoma (HLMC) implanted in athymic mice and on its uptake of human low density lipoproteins labeled with 99mTc (99mTc-LDL). Mice bearing the HLMC were divided into two experimental groups. One of them was administered a control diet (C diet) and the other one was given a diet supplemented with 25 mg GLA/g pellet (GLA diet) for three weeks (Table 1). A tumor growth inhibition with the GLA diet was evident at the second week of treatment, and a marked inhibition (56%) was reached at the end of the third week (Fig. 1). The GLA diet produced some changes in the total fatty acid composition of tumor, plasma and liver of host mice: GLA and arachidonic acid (20:4 n-6, AA) induced significant increases, whereas oleic (18:1 n-9, OA) and linoleic acids (18:2 n-6, LA) were decreased (Table 2). Tumors of those animals fed both diets were labeled by 99mTc-LDL, and no difference was observed in the ratio of tumor/liver and tumor/kidney uptake of host animal (Table 3). Results obtained using this experimental model suggest that the inhibitory effect of GLA on tumor growth is not related to the LDL tumor uptake. PMID- 8731578 TI - Fine needle aspiration for flow cytometry immunophenotyping of non Hodgkin lymphoma. AB - The aim was to evaluate the usefulness of lymph node biopsies obtained by fine needle aspiration (FNA) for immunophenotyping of non Hodgkin lymphoma (NHL). Seventeen superficial and deep lymph node samples were fractioned for conventional cytological examination and immunophenotyping studies. Out of ten NHL, nine were readily detected by flow cytometry (FC), while failure on the remaining case was due to selective loss of large cell population, which is liable to occur with this procedure. A single case, which proved negative for all markers employed, was finally diagnosed by immunohistochemistry as germ cell tumor. The other six cases, presenting lymphoid population without phenotypic abnormalities, were diagnosed by cytology and/or histology as Hodgkin disease or hyperplasic disorders. To conclude, FC immunophenotyping seems to improve the efficacy of FNA in NHL diagnosis, whereas for Hodgkin disease and hyperplasic disorders, classic morphological criteria are more useful for differential diagnosis. Although FNA for FC immunophenotyping cannot replace histopathological examination for NHL diagnosis, it proves to be a useful tool for staging and follow up, making surgical procedures for sample collection unnecessary. PMID- 8731579 TI - Characterization of clinical isolates of Streptococcus agalactiae by random amplified polymorphic DNA using degenerate oligonucleotides. AB - Epidemiological studies of Streptococcus agalactiae strains have been limited by the lack of sensitive and discriminatory methods for comparing clinical isolates. Serotyping, albeit a widely used methodology, has been shown to possess low capability to distinguish between epidemiologically related and unrelated isolates. We have employed here a random amplification of polymorphic DNA (RAPD) assay, using degenerate oligonucleotides as primers, to characterize S. agalactiae isolates from related or unrelated clinical samples. Epidemiologically related isolates (mother-infant pairs) showed identical profiles by this methodology. On the contrary, 12 epidemiologically-unrelated isolates (classified into 5 different serotypes) resulted in 11 distinct RAPD patterns. This suggests that the proposed modified RAPD assay provides a highly discriminatory tool for the analysis of genomic diversity among isolates from pathogenic organisms. PMID- 8731580 TI - [Diarrhea due to microsporidia in a patient with AIDS]. AB - A 26-year-old male AIDS patient with diarrhea of two-months evolution is reported here. The most relevant intestinal pathogens, including Cryptosporidium parvum, were ruled out by routine microbiological tests. Stool samples stained with an "oblong" Ziehl-Neelsen method (fucsin, 7 min instead of 3 min) allowed visualization of organisms resembling microsporidia. Both modified trichrome and calcofluor stains showed organisms compatible with Enterocytozoon bieneusi. Significant titer of antibodies (> or = 1/800) against 4 different microsporidial species were obtained from the serum of the patient by an ELISA test. Clinical improvement was observed after treatment with albendazole, 400 mg twice daily for 4 weeks, even though microsporidial spores were still detected in stool specimens. To our knowledge, this is the first microsporidial infection reported in Argentina. PMID- 8731581 TI - [Isolated neurofibroma of the common bile duct]. AB - The neurogenic tumors in the biliary tract are rare and usually are amputation neuroma that occur after cholecystectomy. We describe a case of isolated neurofibroma of the common bile duct in a young man not cholecystectomized. The patient suffered recurrent episodes of abdominal pain, vomiting and weight loss without clinical signs of Von Recklinghausen's disease or jaundice. The hepatogram was normal. The echography indicated a solid formation with obstruction of the proximal common bile duct. In the ERCP the stenosis was found. Surgical excision of the tumor and anastomosis of bilateral hepatic ducts and jejunum were carried out. At microscopic examination intraparietal neurofibroma of the common bile duct was found. As isolated entity, we know of only one reported case. PMID- 8731582 TI - [Acquired sideroblastic anemia and cholestasis in a hyperthyroid patient treated with methimazole and atenolol]. AB - The authors describe a 62 year-old white male who was diagnosed as autoimmune hyperthyroidism and treated with methimazole and atenolol. Ten days later he showed itching, jaundice and choluria. All drugs were discontinued. The patient was given radioactive iodine. Two months later direct serum bilirubin levels reached 35 mg%. Endoscopic retrograde cholangiogram evidenced normal extrahepatic biliary ducts. The percutaneous liver biopsy showed marked cholestasis specially in the centrolobular zone with a slight infiltrate of mononuclear cells in the portal areas. Together with the liver disease the patient presented an anemic syndrome. Bone marrow aspiration showed rich cellularity, Perls staining showed 70% sideroblasts, with 10% ringed sideroblasts and increased extracorpuscular iron. The patient's evolution was satisfactory. Twenty months after the beginning of the disease clinical and biochemical tests were normal. A new bone marrow aspiration rendered normal. Hepatic cholestasis suffered by our patient was probably due to an adverse reaction of methimazole. Physiopathology of reversible sideroblastic anemia is discussed. PMID- 8731583 TI - [Hemolytic anemia by cryoagglutinins, leukopenia and repetead infections]. PMID- 8731584 TI - [Molecular sex determination. Significance in the diagnosis of gonadal pathologies]. AB - Sex differentiation follows two steps: sex determination where gender follows from the development of the embryonic gonads such as testes or ovaries and sex differentiation leading to the formation of internal and external genitalia. In man, testis determination is dependent on SRY gene (sex region of Y) on the short arm of Y chromosome (Yp11.3). It has an open reading frame that encodes a 220 aa protein with a 80 aa motif related to HMG box. These sequences have been shown to mediate DNA binding. Recently, a DSS locus (Dosage Sensitive Sex-Reversal) has been isolated on Xp21. This gene could be involved in ovaric determination. Moreover, it has been shown that SRY gene expression activates a regulatory pathway that leads to the expression of MIS (Mullerian Inhibiting Substance). Finally, the molecular genetic techniques have provided the tools for the analysis of these genes in patients with gonadal dysgenesis. The information obtained together with phenotype and cytogenetic data has established important correlations of diagnosic value. PMID- 8731585 TI - [100 years of the fantastic X-rays]. PMID- 8731586 TI - [Nobel prize of physiology or medicine 1955: Edward B. Lewis, Christiane Nusslein Volhard, Eric Wieschaus. The flies and the keys of the embryonic development]. PMID- 8731587 TI - [Homo stultus]. PMID- 8731588 TI - [Coronary repermeabilization in acute myocardial infarction]. PMID- 8731589 TI - [Carlos Gianantonio (1926-1995)]. PMID- 8731590 TI - [Esteban Montuori (1906-1995)]. PMID- 8731591 TI - [Nebulization with human IGA in patients with hypogammaglobulinemia and airway infections secondary to lymphoproliferative diseases]. PMID- 8731592 TI - [A weak spot in the immunoprevention of tetanus]. PMID- 8731593 TI - [Recombinant interferon alfa in the treatment of chronic hepatitis C]. AB - BACKGROUND: Chronic hepatitis C usually progresses to cirrhosis and hepatocarcinoma. Treatment with antiviral drugs is indicated attempting to modify the evolution of the disease. OBJECTIVE: To evaluate the treatment of chronic hepatitis C or post-hepatitis C cirrhosis with interferon-a-R (IFN) and to identify the factors associated with good therapeutic result. METHODS: Thirty eight patients with chronic hepatitis C or post-hepatitis C cirrhosis were treated with 2.5 or 3.0 MU of IFN three times a week for 6 to 12 months. We considered as a complete and sustained response when ALT and AST were normal for a 6 months period after finishing treatment. We considered complete response with relapse those in whom elevation of the enzymes was detected after drug suspension. RESULTS: There was normalization of ALT and AST in 17 out of 38 patients (44%). In this group, 9 out of 17 presented a complete and sustained response and in 8 out of 17 there was elevation of the enzymes after drug interruption. There was a trend of better response to interferon in patients below 40 years of age and in those with chronic active hepatitis. The most frequent side effects were fever (80%), myalgia (60%), asthenia (50%), headache (40%), and arthralgia (36%). CONCLUSION: Treatment with interferon showed a satisfactory and sustained response in 23% of the subjects, with better results in young patients and without cirrhosis. PMID- 8731594 TI - [Serotypes of classical enteropathogenic Escherichia coli that produce changes in the small bowel ultrastructure and invasion of HeLa cells]. AB - Enteropathogenic Escherichia coli (EPEC) is the main cause of diarrhea in infants up to one year of age in the majority of the developing countries. In vitro EPEC strains attach to HeLa or HEp-2 cells in a specific pattern called localized adherence (LA), which is correlated with 93% of the EPEC serotypes. In vivo, EPEC strains adhere intimately to cuplike projections of the apical enterocyte membrane causing localized destruction of the microvilli, described as an attaching and effacing lesion. PURPOSE: We showed the attaching-effacing lesions and intracellular penetration in the ultrastructural study of the small intestinal cells as well as in the HeLa cell assay by the following EPEC serotypes: O111ab:H2, O119:H6 and O18ab:H14. PATIENTS AND METHODS: These strains were isolated from the stools of three infants less than 2 years of age with watery diarrhea who were hospitalized for fluid and electrolyte repairment. In the three patients there were severe ultrastructural alterations of the enterocytes mainly shortening and destruction of the microvilli, and formation of cuplike pedestal lesions. There was also penetration of microorganisms into the cytoplasm of the enterocytes in the interior of endocytotic vesicles. The serotypes of EPEC were assayed with HeLa cells showing the formation of pedestals and penetration into the cytoplasm. CONCLUSION: The ultrastructural alterations of the small bowel observed in these patients represent an important indication of the possibility of perpetuation of the diarrhea, owing to either a secretory mechanism or malabsorption of the nutrients. PMID- 8731595 TI - [Influence of bile salts on the motor response of isolated ileum to acetylcholine, in rats]. AB - Absence of bile salts in the intestinal lumen of jaundiced patients is associated to bacterial overgrowth and systemic endotoxemia. These bile salts, however, did not show significant influence on aerobic and facultative intestinal bacteria. The increasing bacterial colonization may be due to depressed intestinal motor response. PURPOSE: To evaluate the influence of bile salts on intestinal motor response in presence of obstructive jaundice. METHODS: We studied in vitro the motor response of ileal segments of 30 Holtzman rats divided into three groups (n = 10): washed ileum, intraluminal bile salts and exogenous oral bile salts administred during six days. Five animals of each group were submitted to sham operation and the other five were submitted to ligation and section of the common bile duct. A four centimeter ileal segment was isolated and studied through a dose-response assay with acetylcholine in an organ chamber. RESULTS: The results showed an increased ileal affinity to acetylcholine in presence of intraluminal bile salts. CONCLUSION: The intraluminal bile salts appear to exert in vitro a stimulatory effect on ileal motility. PMID- 8731596 TI - [Value of CA 125 as a tumor marker in ovarian carcinoma]. AB - Cancer antigen 125 (CA 125) is a high molecular weight antigenic glycoprotein. It is recognized in the serum by a monoclonal antibody. Rising of serum levels is most frequently observed in borderline and malignant serous ovary tumors. Occasionally other cancers show elevated levels of CA 125. Structure derived from the celomic epithelium can also show elevated levels of CA 125. PURPOSE: Review the useful of CA 125 for diagnosis and follow-up of patients with ovarian carcinoma. METHOD: To obtain the information, an extensive and up-to-date bibliographic review was done about this topic. RESULTS: This tumor marker has an excelent sensibility although it is not especific. As it cannot develop tumors smaller than 1 cm, it is not indicated for populational screenings. CONCLUSION: CA 125 is useful to follow-up of patients whose ovarian carcinoma had already been diagnosed and suffered surgery procedures followed for chemotherapy. In this cases CA 125 show the therapeutic response. PMID- 8731597 TI - [Epidemiologic analysis of genital diseases associated to papillomavirus]. AB - In order to investigate genital papillomavirus infection in a university hospital, we verified that condyloma acuminata and epidermoid carcinoma were the most common diseases associated to this virus. PURPOSE: This study was undertaken to verify the prevalence of genital lesions associated to papillomavirus and sex, age and race distribution. METHODS: We have reported 223 patients who were treated at Hospital Universitario Antonio Pedro, Rio de Janeiro State, during the period 1988-1992. After clinical diagnosis, biopsies were submitted to histological analysis. RESULTS: Results showed that condyloma acuminata and epidermoid carcinoma were the most frequent lesions among genital lesions. Carcinoma cases had a stable prevalence along the period while condyloma cases had a significant increase. Both lesions were more frequently found in Caucasian (white) patients. There were no differences between sexes and condyloma evidence. The media age in the epidermoid carcinoma cases did not change in the years, but it has decreased regarding condyloma acuminata. CONCLUSION: Genital diseases associated to papillomavirus increased during five years. Attention must be paid to control and early diagnosis to prevent neoplasic evolution. PMID- 8731599 TI - [Evaluation of deaths by accidents with venomous snakes in the State of Sao Paulo, 1988/1989]. AB - In Brazil, the discussion about the quality of medical care evaluation is beginning. PURPOSE: Assessment of the cases of death by venomous snake bite in the State of Sao Paulo. METHODS: Twenty one deaths caused by venomous snakebite, that occurred in the State of S-ao Paulo, Brazil, during 1988 and 1989 were analysed. The method of three audits implicit criteria was used to evaluate the care given to these cases. Two of the authors did also a qualitative analysis of the care. RESULTS: The results showed a higher frequency of males and adults of 50 and more years of age. The answers to the question "if the death was predictable" were all negative, in 5 cases, and the majority agreed in 6 cases (52.4% of answers were "not predictable"). The question "if there was evidence that the complication was recognized" had answers as "yes" or "there was not sufficient information" in the majority. As to the question "all the appropriate measures were taken in view of the complications", in three cases the answer was "no". CONCLUSION: The main question mentioned was the bad quality of the information in the medical records. The qualitative analysis of the two authors, showed, some inadequate procedures. PMID- 8731598 TI - [Gastroesophageal reflux in children. Surgical treatment by the Nissen's technique]. AB - BACKGROUND: To analyse the results of Nissen's fundoplication in the treatment of gastro-esophageal reflux (GER) in children. MATERIAL AND METHODS: A retrospective study was undertaken in 185 children with GER, diagnosed through upper gastrointestinal (GI) tract barium examination, and admitted at Hospital Infantil Joana de Gusmao-Florianopolis, SC, Brazil, from February 1981 to June 1989. From the charts the following data were reviewed: age, sex, clinical picture, subsidiary tests, surgical indications, surgical treatment by Nissen's fundoplication, pre and transoperative treatment, post operative complications and results classified as good (no reflux), bad (relapse), unknow (no follow-up) and deaths. RESULTS: The age ranged from 30 days to seven years and sex incidence was predominant in males (2:1). The main clinical manifestation were vomits (85.40%), respiratory distress (64.32%) and malnutrition (55.13%). Upper GI tract examination revealed isolated GER (40.00%), associated to hiatal hernia (18.91%) and complicated with esophagitis (29.18%) and peptic stenosis (11.89%). In the transoperative period there was terminal esophagus perforation in one case. The main postoperative complications were gas bloat syndrome (7.02%) and edema of the terminal esophagus (5.94%). The result was good in 159 children (85.94%), bad in 7 (3.78%) and unknown in 11 (5.94%). There were 8 deaths (4.32%): 1 (0.54%) related to the disease, 1 (0.54%) to the surgery and 6 (3.24%) related to medical causes. CONCLUSIONS: Based on these results, Nissen's fundoplication is an adequated operation for treatment of children with GER. PMID- 8731600 TI - [Tissue expansion of the superior limb]. AB - In order to get better results in the treatment of the superior limbs defects we have attempted to take profit of the expansion tissue techniqu. PURPOSE: With the objective of studying the utilization of expansion tissue technique to repair defects in the superior limbs caused by scars or tattoos removal. METHODS: 6 patients with 7 superior limbs defects to be repaired were treated by the technique of tissue expansion with 8 tissue expanders. In 3 patients the lesions were caused by tattoos removal (4 lesions), 2 patients presented lesions caused by burn scars removal nd 1 patient showed lesion caused by removal of a childhood scar. RESULTS: The result of this procedure was excellent in 5 cases and provided total repair of the lesion. However hazards have occurred in one patient who showed a necrosis of the expansion neighbouring area (not a consequence of the expansion) and in other patient the aesthetic result was not satisfactory because he developed a hypertrophic scar. Good aesthetic results were obtained in the other 4 patients. CONCLUSIONS: We have had good results with the utilization of tissue expanders in the superior extremities. The superior limbs have good capacity of expansion and lead to good aesthetic results. Any significant hazards were observed as being a consequence of superior limb tissue expansion. PMID- 8731602 TI - [Heterogeneity in bibliographic references]. AB - Bibliography plays an important role in the writing of books and papers, as it permits us an exchange of experience, the reading of case reports, the revision of articles and new investigative aspects. Bibliographic references in the same chapter of six books from different countries, published in the same period, were analyzed, and a strong heterogeneity was found among them. The "language pattern" varied depending on the author's native language. The temporal distribution of articles was also different and only a small percentage of articles was common to two or more books. Our results showed that the same chapter in different books have highly heterogenous bibliographic references, although they should have similar contents. PMID- 8731601 TI - [Prevalence of diabetic nephropathy in adult patients with chronic kidney failure]. AB - Diabetic nephropathy is a frequent cause of end-stage renal failure in patients admitted for renal replacement therapy. PURPOSE: To evaluate the prevalence of DN, as the underline disease, in patients with ESRF. METHODS: 1,303 [male (M) = 767 and female (F) = 536] patients with ESRF who were on a waiting list for cadaver kidney transplant at Nephrology Unit-University Hospital (HC-UNICAMP), from August/90 to June/93--group 1--and 193 (M = 112 and F = 81) patients admitted for renal replacement therapy in a year period (April/92 to March/93), in the city of Campinas, State of Sao Paulo, Brazil, were studied. RESULTS: The prevalence of DN was 10.1% in group 1 and 17.6% in group 2 (x2 = 7.15; p = 0.007), being the third cause of ESRF in both groups, and it was preceded by glomerulonephritis and arterial hypertension. In group 1 the reduction of number of patients with increase in duration of dialysis was significantly greater in patients with diabetic nephropathy (x2 = 30.9; p < 0.001). Among patients with DN 35 (26%) in group 1 and 6 (18%) in group 2 had less than 35 years when they were admitted for renal replacement therapy and are likely to be type 1 (insulin dependent) diabetic patients. CONCLUSION: In our studied groups DN was a frequent cause of ESRF. PMID- 8731603 TI - [Esophageal injury due to pill ingestion]. AB - In recent years, many case reports concerning esophageal injuries caused by drugs have been published. The primary cause has apparently been the delay in passage and the adherence of the caustic drugs on the esophageal mucosa. The authors report a case of esophageal ulceration caused by an analgesic in a 26-year-old male with no esophageal symptoms. A review of the literature shows that a variety of medications have been implicated; size, shape and improper ingestion of pills affect esophageal transit. PMID- 8731604 TI - [The plastic surgery in a multidisciplinary context and future perspectives]. PMID- 8731605 TI - [Clinical and epidemiological characteristics of acute diarrhea by classical enteropathogenic Escherichia coli]. AB - Acute diarrhea is usually considered as a self limited disease, but under certain circumstances, mainly owing to the age of the patient, the nutritional status and the enteropathogenic agent the illness can evolve for a protracted evolution. PURPOSE: In the present study we report the clinical and epidemiological features of a group of infants under two years of age with acute diarrhea caused by serogroups of enteropathogenic Escherichia coli (EPEC). PATIENTS AND METHODS: During a two year period 200 infants under two years of age, mean 8.2 months, with acute diarrhea less than 5 days duration, were consecutively studied. A control group of 40 healthy infants matched for age was also made up. The nutritional status was determined and the occurrence of food intolerance was also monitored. The patients were followed up for 4 weeks after been discharged from the hospital. Stool samples were obtained for research of bacterial, viral and protozoan enteropathogenic agents. RESULTS: EPEC was isolated in the stools of 84 (42.0%) infants, as a sole enteropathogenic agent in 55 (27.5%) and in the remaining 29 (14.5%) infants associated with some other agent. EPEC was isolated in the stools of 9 (22.5%) infants of the control group (p < 0.05). Food intolerance was the main digestive complication and also the most important factor that caused perpetuation of diarrhea. The mean duration of the disease was 11.2 days, varying from 2 to 40 days. In 53 (71.6%) infants the disease lasted less than 14 days, while in the remaining 21 (28.4%) it lasted more than 14 days, and all these infants presented with food intolerance. CONCLUSIONS: Enteric infections caused by EPEC serogroups, particularly O111 and O119, are more prevalent in weaned infants under 1 year of age belonging to families of low income rates. These infants present heavy fluid and electrolyte losses in the stools associated to high rates of food intolerance. PMID- 8731606 TI - [Esophageal squamous cell papilloma. Report of 3 cases. Diagnostic aspects]. AB - Dysphagia is not always present in patients with esophageal squamous cell papilloma, and it can be an incidental finding during gastrointestinal endoscopy of an asymptomatic patient. PURPOSE: Three cases of squamous cell papilloma of the esophagus and aspects regarding diagnosis and association with human papillomavirus are presented. CASUISTIC: Two cases are male and one female what is consistent with a higher incidence showed in the literature in males. The three patients were submitted to upper gastrointestinal endoscopy: in one case the tumor was unusually large and in all patients it was located in the mid and lower esophagus. Chromoscopy was performed in one case and the lesion was not stained. We were unable to identify human papillomavirus using DNA-hybridization techniques. Our results are similar to those reported in the review of the literature. RESULTS: There is no other case published in the Brazilian literature and this is the first report on the use of chromoscopy. CONCLUSION: We were not able to show an association between esophageal squamous papilloma and human papillomavirus. PMID- 8731607 TI - [Severe bradyarrythmia induced by tricyclic antidepressants in an elderly patient]. AB - A 66 year-old female patient, with severe chronic pain caused by diabetic radiculopathy, treated with association of amitriptiline and carbamazepine, presented severe sinusal bradycardia with hemodynamic disturbances ensuing junctional rhythm, which required installation of temporary pace-maker. The drugs were withdrawn. Reversion to sinusal rhythm occurred 36 hours after the installation of bradycardia. The pace-maker was withdrawn. It was concluded that the use of tricyclic antidepressive drugs, particularly in the elderly, should be carefully monitored. PMID- 8731608 TI - ["Mercy lie" for the patient with cancer]. AB - The authors discussed the problem of whether or not to be completely frank with cancer patients about their diagnosis. They analysed the results of a survey of 79 doctors (Phase 1) in which they tried to find out how the doctors behaved towards the patients, their families and with themselves in the different situations caused by the illness. In the second stage (Phase 2), they also put the following question to 118 adults: If it was diagnosed that you had cancer would you like your doctor to tell you? PMID- 8731609 TI - [Historical evolution of the treatment of esophageal cancer at the Hospital A. C. Camargo, Sao Paulo, Brazil]. AB - OBJECTIVES: From 1947-1986 we reviewed a historical series of 1,900 cases of esophageal cancers registered at the A.C. Camargo Hospital, Sao Paulo, Brazil. Two hundred and thirty four cases were submitted to surgical resection. During these 4 decades the treatment philosophy of these tumors has changed. METHOD: Five different historical groups were identified and the results are presented. RESULTS: The first group (1947-60) consisted of 47 cases only submitted to surgical resection. The second group (1961-70) of 56 cases had pre and pos surgery radiotherapy in low doses and the reconstruction was made using subcutaneous colon. From 1971-75 the same approach was used except with high dose preoperative radiotherapy (31 cases). In the 4th group (1976-82) of 68 cases preoperative radiotherapy (high dose) and chemotherapy were used. In the last group (1983-86) composed of 32 cases the treatment was preoperative chemotherapy, surgical resection with gastric reconstruction followed with high doses radiotherapy in the surgical bed and chemotherapy. The only significant prognostic factors in the statistical analysis were tumor size and involvement peri-esophageal lymph nodes. CONCLUSION: A five year survival from 3.7% to 9.0% was obtained through the use of the fifth group treatment planning. PMID- 8731610 TI - [Xenotransplantation]. PMID- 8731611 TI - [Pharmacology, clinical indications and toxicity of modern fluoroquinolones]. PMID- 8731612 TI - [Neurophysiological basis of acupuncture]. PMID- 8731613 TI - [Cardiovascular effects of aqueous extracts of the leaves of Clusia coclensis (Guttiferae)]. AB - Aqueous extracts of the leaves of Clusia coclensis, (Guttiferae) injected intravenously to 42 normal Sprague-Dawley rats and to 42 Spontaneous Hypertensive rats, in 7 different doses, induced a rapid and transitory decrease in blood pressure and heart frequency. The magnitude of blood pressure decrease was dose dependent. The effect was statistically significant (p < or = 0.01), probably due to sympathetic stimuli. PMID- 8731614 TI - [Diuretic effect of Cecropia obtusifolia (Moraceae) on albino rats]. AB - The possible diuretic effect of the leaf extract of Cecropia obtusifolia, Bertol; used in Costa Rican traditional medicine, was tested on five Sprague Dawley rats in individual metabolic cages. During the control week, distilled water was administered with an intragastric cannula. During the C.obtusifolia week, the rats were given the aqueous leaf extract in a daily dosage of 500 mg/kg, intragastricly. Although, the urinary flow was increased (p < 0.05) by a 20%, there was no statistically significant difference between the following parameters: excretion in urine of osmoles, sodium and potassium; daily ingestion of food and water. The animal weights decreased when C. obtusifolia was used, but only on the fourth day the decrease was significant (p < 0.05). In conclusion, the aqueous leaf extract of C. obtusifolia has a slight diuretic effect. PMID- 8731615 TI - [Susceptibility of Sigmodon hispidus (Rodentia: Cricetidae) to Toxoplasma gondii (Eucoccidia: Sarcocystidae)]. AB - Sigmodon hispidus, a cotton rat, was inoculated with tachyzoites or oocysts of Costa Rican strains of Toxoplasma gondii to demonstrate the resistance to this parasite, as compared with mice, hamster and white rat infection. Susceptibility to tachyzoite inoculation was higher in this animal than in white rats but lower than in the other animals. Independent to the Toxoplasma strain studied, oocyst infection was less lethal for S. hispidus when compared with mice and hamsters; the results were similar to those observed for white rats. There is a probable participation of this wild animal in the natural Toxoplasma life cycle. PMID- 8731616 TI - Content of a trainer's report for summative assessment in general practice: views of trainers. AB - BACKGROUND: Summative assessment of doctors completing general practice vocational training will become compulsory for those completing training after 1 September 1996. One component of the assessment will be a trainer's report. AIM: A study set out to consider the content of the trainer's report. It aimed to determine which elements of general practice general practitioner trainers believe are the most important for independent general practice; to seek the views of trainers as to the most appropriate methods for assessing these elements in summative assessment of general practitioner registrars; and to determine how frequently trainers have been sufficiently concerned about the performance of registrars to consider not signing the form denoting satisfactory completion of the training year. METHOD: A questionnaire was sent to 1296 general practitioner trainers in the United Kingdom. The main outcome measures used were: the percentage of respondents indicating that an element was very important/crucial for independent general practice; the percentage of respondents indicating a favoured method of assessment that did not include a trainer's report at all; and the proportion of trainers who had ever considered not signing the form denoting satisfactory completion of the training year. RESULTS: The response rate was 78%. Of 75 elements examined, 31 were considered important for independent general practice by 70% or more of respondents. For 29 of these 31 elements, fewer than 30% of respondents would choose a method of assessment that did not include a trainer's report at all. Twenty-six per cent of respondents had considered not signing the form denoting satisfactory completion of the training year, which represents a trainer considering not signing this form once every 29 years. CONCLUSION: These results provide a basis for the content of a structured trainer's report for summative assessment in general practice; such a report is likely to contain about 30 items. Data from the Joint Committee on Postgraduate Training for General Practice and the present study suggest that, for every form not signed, trainers considered not signing the forms of another 13 general practitioner registrars. This highlights the need for a report that will help trainers to make the difficult decision as to whether or not a registrar is ready for independent practice. PMID- 8731617 TI - Consensus minimum standards for use in a trainer's report for summative assessment in general practice. AB - BACKGROUND: Summative assessment of general practitioner registrars is to be introduced in September 1996, one component of which will be a report by the trainer. Standards must be set and guidance provided as to the most appropriate ways of obtaining evidence for the trainer's assessment. AIM: The first aim of this study was to set consensus minimum standards for 30 items that are likely to form the content of a trainer's report; the second aim was to provide a consensus view on the most appropriate methods of assessment to be used by trainers. METHOD: A consensus conference was held in March 1995 during which the 30 items were discussed by a group of 30 general practitioners, of whom 27 were experienced trainers. This resulted in a draft document that was circulated to the conference attenders and other experts for consultation. RESULTS: Draft minimum standards were produced for all 30 items after the consensus conference with a mean of 2.5 standards for each item. Of those involved in the consultation exercise, 82% replied. Most of the revisions suggested at this stage were of a minor nature; the only major revision was to divide one item into two, resulting in a final total of 31 items. All but one of the 80 standards could be assessed by direct observation; 41 (51%) could be assessed by tutorial-based discussion and 61 (76%) by methods specific to that standard. Trainers or their practice partners were viewed as acceptable sources of evidence for all items and hospital consultants and primary health care team members were viewed as acceptable for just over half of the items. CONCLUSION: Standards for use by trainers when providing a general practitioner report for the summative assessment of registrars have been developed by consensus conference and have been subjected to review by consultation. Acceptable methods by which registrars could be assessed against these standards, and suitable personnel who could provide evidence, have also been suggested. PMID- 8731618 TI - Which smokers are helped to give up smoking using transdermal nicotine patches? Results from a randomized, double-blind, placebo-controlled trial. AB - BACKGROUND: Nicotine replacement therapy is effective in helping people to give up smoking. The three forms now available--transdermal patches, chewing gum and nasal spray--deliver nicotine at different rates and to different levels. Therefore, it might be expected that smokers with different characteristics, and at different levels of nicotine dependence, will be helped more by one or other method. AIM: The aim of the study was to examine whether the effectiveness of transdermal patches is related to nicotine dependence or to other smoker characteristics and to investigate factors predicting smoking cessation using patches. METHOD: Data from a randomized, double-blind, placebo-controlled trial of nicotine transdermal patches were analysed retrospectively. The trial, conducted in 1990-1992, involved 1686 patients recruited from 19 general practices in Oxfordshire. The main outcome measure was continuous smoking cessation from 8 to 52 weeks after the start of patch use, biochemically validated at 12, 24 and 52 weeks. The effectiveness of the patches was measured by the relative odds of sustained cessation using nicotine patches compared with placebo patches. RESULTS: Nicotine transdermal patches were more effective in smokers with moderate nicotine dependence [odds ratio (OR) 1.94; 95% confidence interval (CI) 1.24-3.04] than in mildly or highly dependent smokers (OR 0.98; 95% CI 0.58-1.65) (difference in ORs P < 0.05) and more effective in those aged 24-49 years (OR 1.89; 95% CI 1.24-2.87) than in older smokers aged 50-65 years (OR 0.88; 95% CI 0.49-1.59) (difference in ORs P < 0.05). Abstinence from smoking in the first week of the trial was the strongest predictor of sustained cessation and was more common among smokers using nicotine patches than those using placebo patches (33% of 842 compared with 22% of 844; P < 0.001). Of first-week abstainers, 25 and 28% of 277 and 182 in the nicotine and placebo groups, respectively, achieved sustained cessation compared with 4% of 565 and 2% of 662 first-week smokers. CONCLUSION: Nicotine transdermal patches were most effective for smokers with moderate nicotine dependence and for younger smokers. Early abstinence from smoking was the strongest predictor of sustained cessation. A week's trial of the patch proceeding to longer term use if abstinence is achieved may be an effective policy. PMID- 8731619 TI - Characteristics of drug misusers and their perceptions of general practitioner care. AB - BACKGROUND: Little is known about drug misusers' views of the care they receive from general practitioners. AIM: This study set out to determine drug users' views about primary health care and their relationship with their general practitioners. METHOD: A semi-structured interview was conducted with 180 drug users who were consecutive attenders at five treatment services in north east London--a general practice with a special interest in treating drug users, a private drug clinic, a community drug team, a drug dependence unit and a street agency for drug users. RESULTS: The majority of the 145 London-based drug users attending the four treatment centres other than the general practice with a special interest were registered with a general practitioner (88%). Forty-two per cent of users sought out a general practitioner prepared to treat them, rather than register with a local or the family doctor. Most drug users reported that their general practitioners were aware of their drug problems (88%) but half of the general practitioners were not prescribing replacement drugs with almost 20% of the users not receiving prescriptions claiming that this was due to lack of knowledge or trust on the part of the doctor. Sixty per cent of the drug users attending the four centres perceived that their general practitioners held negative or neutral views about them. However, 34 of the 35 drug users interviewed in the specialist general practice believed their doctors had a positive view of drug users. CONCLUSION: Most drug users were registered with general practitioners but the relationship between doctor and patient was not always easy or productive. There is a need to clarify the role of general practitioners in this field and provide them with better educational opportunities. PMID- 8731620 TI - Patients' views of low back pain and its management in general practice. AB - BACKGROUND: Low back pain is a common and persistent problem. Research studies seeking to improve the quality of management of this condition have tended to ignore the opinions of patients. There is a growing acceptance of the importance of taking patients' views into account in developing management and educational programmes for a variety of conditions. AIM: This study set out to elicit the views of patients concerning low back pain and its management in general practice. METHOD: Fifty-two in-depth interviews were conducted with patients selected from a broad range of 12 general practices. RESULTS: Analysis of the interviews identified seven themes relating to: quality of life, prognosis, secondary prevention, help-seeking behaviour, explanation of underlying pathology, satisfaction with general practitioner management, and complementary therapy. Different patient viewpoints or perspectives were expressed within each of these themes. Patients adapted to the progress of their low back pain and were not seeking a 'magical cure' from either conventional or complementary therapies. CONCLUSION: Patients' views on low back pain are heterogeneous. The dissatisfaction expressed with medical explanations for the pain may be related to superficial clinical management and the constraints of general practice. Good management of low back pain needs to take patients' complex views of the condition into account. PMID- 8731621 TI - Stress among general practitioners and their spouses: a qualitative study. AB - BACKGROUND: Although research has been carried out on stress in general practitioners, little is known about the stresses experienced by their spouses. AIM: This study was undertaken to identify specific pressures at work and at home experienced by general practitioners and their spouses and to highlight their coping strategies. METHOD: In-depth interviews were conducted with 25 general practitioners and their spouses in the north-west of England. RESULTS: The doctors' increased workload and decreased interest in their family are important stressors for the entire family unit. Other stressors include time pressure, out of-hours on-call, lack of support and amount of paperwork. General practitioners work late in the surgery, bring work home and spend time away from home at meetings. Family life is constantly interrupted by telephone calls. Role conflict was one of the major sources of stress for women general practitioners. Doctors' wives expressed concern regarding their husbands' excessive commitment to work and problems with communication. CONCLUSION: Recent changes to contractual working conditions have caused general practitioners to focus their energies on their practice instead of paying attention to the needs of their family. Male general practitioners leave the bulk of responsibility for running the family and household to their wives, but women general practitioners appear to maintain domestic responsibility while spending as much time in medical practice as their male colleagues. This is only an exploratory study based on a small sample, and so the findings cannot be generalized; however, it provides in-depth information on stress among general practitioners and their spouses. PMID- 8731622 TI - Incidence and causes of rectal bleeding in general practice as detected by colonoscopy. AB - BACKGROUND: Rectal bleeding is common, but it is still unclear which patients require investigation to exclude serious pathology, although it is known that colectoral cancer is very rare under the age of 40 years. Few studies have examined all patients presenting to their primary health physician rather than screening whole populations. AIM: The aim of this study was to investigate the view that all patients over the age of 40 who present to their general practitioner with rectal bleeding should undergo investigation by colonoscopy to rule out serious pathology, regardless of symptomatology. METHOD: A prospective study was carried out of 99 consecutive patients over 40 years presenting with rectal bleeding to 17 general practices in Newcastle upon Tyne. RESULTS: Serious pathology was detected by colonoscopy in 44.4% of patients. The diagnoses were: colorectal carcinoma, eight cases (two Dukes' grade A, two Dukes' grade B, four Dukes' grade C); one or more polyps, 25 cases (in 17 cases at least one polyp was 5 mm or greater in diameter); inflammatory bowel disease, 11 cases. In the remaining 55 patients, bleeding was associated with diverticular disease (16 cases) and haemorrhoids (28 cases). No cause was found in 11 patients. This high rate of pathology may be partly caused by selection of cases for referral by the general practitioner, despite efforts to minimize this. Three symptoms as elicited by the colonoscopist were found to be significantly associated with serious disease: blood mixed with stool (P < 0.001); change in bowel habit (P < 0.005); and the presence of abdominal pain (P < 0.025). However, symptoms elicited on primary presentation were less helpful and symptoms changed significantly between consultation with the general practitioner and colonoscopy. CONCLUSION: All patients over the age of 40 years presenting with rectal bleeding should be referred for flexible sigmoidoscopy or colonoscopy. Symptoms are unhelpful in deciding who requires investigation. PMID- 8731623 TI - Comparison between primary care physiotherapy education/advice clinics and traditional hospital based physiotherapy treatment: a randomized trial. AB - BACKGROUND: Studies have shown that patients prefer to received physiotherapy services in the primary care setting, but none has made direct comparisons between hospital and primary care based physiotherapy. AIM: This pragmatic randomized trial set out to compare general-practice-based physiotherapy education and advice clinics with traditional physiotherapy treatment in an acute hospital setting. METHOD: The study involved 130 patients referred to physiotherapy services by 43 general practitioners over a one-year period. Patients were included in the study if they were at work, independent in all activities of daily living and would have routinely been referred to the hospital physiotherapy department for treatment. Patients were randomly allocated to one of two intervention groups: general practice education/advice, or treatment and education/advice at the local hospital. Patients completed a questionnaire prior to their first physiotherapy appointment and again 6 weeks later. RESULTS: A disappointing number of patients failed to attend either the first or subsequent appointments. The post-intervention scores revealed improvements in patients' problems, with the advice group exhibiting a slightly better outcome than the hospital group, as measured by the Nottingham health profile, the anxiety component of the hospital anxiety and depression rating scale, pain and problem size visual analogue scales, and measures of patient satisfaction. Advice group patients also had less attendances than the hospital treatment group. The general practitioners surveyed commented favourably on the advice clinics. CONCLUSIONS: The findings of this study justify the concept of general-practice-based physiotherapy education and advice clinics. PMID- 8731624 TI - Do self-management plans reduce morbidity in patients with asthma? AB - BACKGROUND: Self-management plans may help patients with asthma intervene when symptoms deteriorate, thus preventing asthma attacks. AIM: A study set out to test whether a self-management plan tailored to the circumstances of the individual reduces morbidity from asthma. METHOD: General practitioners who had participated in a national audit of asthma attacks were randomized into intervention and control groups. Six months after the intervention group had issued self-management plans to patients with asthma, both groups of practitioners completed morbidity questionnaires on patients. Morbidity outcomes were compared for the 6-month periods before and after the issue of the plans. RESULTS: In the 6 months before the study, the 376 patients enrolled by the intervention group experienced higher levels of morbidity than the 530 patients for whom details were recorded by the control group. In the 6 months after the issue of the plans, control group patients showed little change in levels of morbidity, but intervention group patients showed significant reductions in hospital admissions, consultations for asthma symptoms, asthma review consultations, courses of oral steroids and use of emergency nebulized bronchodilators. CONCLUSION: General practitioners appeared to operate enthusiast bias' and issued more self-management plans to patients with uncontrolled asthma. The reduction in morbidity in this group is probably a result of the use of the plans, but the verdict on whether plans reduce morbidity must be deemed 'not proven'. PMID- 8731625 TI - Effects of immediate modified feeding on infantile gastroenteritis. AB - BACKGROUND: Standard treatment of infants who are dehydrated as a result of acute gastroenteritis is to administer oral rehydration therapy (ORT). Traditionally, food has been withdrawn for 24-48 h, but there is no conclusive evidence that this is of any real benefit to the patient. Immediate modified feeding, in which an infant on ORT is not starved but administered a limited diet, may have benefits in the treatment of gastroenteritis, especially in children who are nutritionally compromised before they develop the illness. AIM: A pilot study was carried out to investigate the effects of giving infants suffering from acute gastroenteritis a limited modified diet in conjunction with ORT. METHOD: Infants recruited into the study by their general practitioner or by a research doctor in the hospital casualty unit of Bristol Children's Hospital were randomly allocated to receive ORT with or without immediate modified feeding. The duration of diarrhoea, weight change, and incidence of vomiting and lactose intolerance were measured in both treatment groups, and the results were compared. RESULTS: Of the infants studied, 27 received ORT and immediate modified feeding, and 32 ORT alone. The duration of diarrhoea, and incidence of vomiting or lactose intolerance were no greater in the group receiving immediate modified feeding. Patients who received ORT and immediate modified feeding appeared to gain more weight than the infants who were starved for 24-48 h, but this difference was not statistically significant. CONCLUSION: Immediate modified feeding is safe and effective, and may have nutritional advantages over traditional ORT with starvation. A similar but multicentre study using unmodified diet, i.e. child's normal diet, is being carried out by a working group of The European Society of Paediatrics, Gastroenterology and Nutrition (ESPGAN). PMID- 8731626 TI - The role of screening for Helicobacter pylori in patients with duodenal ulceration in the primary health care setting. AB - BACKGROUND: It is known that at least 90% of duodenal ulcers are caused by infection with the bacterium Helicobacter pylori. Eradicating this organism usually results in complete resolution of the disease. Testing for H pylori was introduced relatively recently, and thus, many patients known to have uncomplicated peptic ulcer disease who continue to need long-term treatment with ulcer-healing drugs have never been tested for the infection or offered eradication therapy. In modern computerized practices, this subgroup of patients can readily be identified by reference to morbidity and repeat prescribing data. Eradication of H pylori infection in this group of patients has great potential benefit for the individuals concerned as well as cost-saving benefit for the National Health Service. AIM: The aim of this prospective study was to determine whether it is worthwhile screening for and treating H pylori infection in patients in a general practice population with previously diagnosed duodenal ulcer disease taking ulcer-healing drugs long term. METHOD: In 1994, in a practice of 7100 patients, morbidity and repeat prescribing data were used to identify 40 patients (0.6%) with proven duodenal ulcer disease taking ulcer healing medication long term and with uncertain H pylori status. Twenty-nine of the 40 subjects agreed to undergo serology testing for H pylori antibodies. Of 20 (69%) who were positive, 18 (eight women, median age 63.8 years) were given eradication therapy. Seventeen patients received omeprazole 40 mg once daily and amoxycillin 500 mg three times daily for 14 days with metronidazole 400 mg three times daily for the first 7 days; for the remaining patient metronidazole was inadvertently omitted. [13C]Urea breath testing was carried out at the local hospital at least one month after therapy to determine whether eradication treatment had been successful. Subjects were also personally followed up by telephone after 1 and 4 months to assess the success of treatment subjectively. RESULTS: [13C]Urea breath testing showed that H pylori eradication was successful in all 17 patients (100%) who received the intended eradication regimen. Helicobacter pylori was not eradicated in the patient who received only omeprazole and amoxycillin. Four months after successful H pylori eradication, 13 of the 17 (76%) patients remained completely asymptomatic. Two of the four patients who had some recurrent dyspepsia had known gastro-oesophageal reflux and their ongoing symptoms after eradication therapy seemed, on close questioning, to be more attributable to this than to duodenal ulcer disease. CONCLUSION: Testing for and eradication of H pylori is worthwhile in general practice in those patients with previous proven duodenal ulceration who need long-term ulcer healing medication. The high rate of eradication of H pylori achieved with the regimen used in this study compares very favourably with that of other treatment regimens. However, in patients with duodenal ulcers there may be coexisting pathology, and H pylori eradication does not necessarily result in complete disappearance of dyspeptic symptoms. Thus, when monitoring the outcome of treatment it is important to assess improvement of symptoms as well as objective evidence of eradication. PMID- 8731627 TI - Completeness and accuracy of morbidity and repeat prescribing records held on general practice computers in Scotland. AB - BACKGROUND: A high proportion of Scottish general practices use a standard computer software package (GPASS, general practice administration system for Scotland), and thus, Scotland is uniquely placed to amalgamate primary care data on a national scale. Practices, however, vary widely in the nature and extent of data entered on computer and a major limitation on the use of the collected data is the absence of information on the completeness and accuracy of the computer database. AIM: This study set out to assess the quality of morbidity and repeat prescribing records held on computer by general practices in Scotland. METHOD: Forty-one practices, with above average levels of morbidity data recorded on computer, were selected on a geographic basis in relation to the national population distribution. Within each practice, 250 patients aged 45-64 years were selected at random. Data relating to 19 diagnoses, six surgical procedures and 40 repeat prescription drugs were extracted from the computer records of these patients and compared with information held on patients' paper records and supplied by patients in response to a postal questionnaire. The completeness and accuracy of computer entries were assessed in terms of sensitivity and positive predictive value, respectively. RESULTS: For the 5567 patients for whom all three sources of data (validated computer records, paper records and questionnaire responses) were available, sensitivity (completeness) of morbidity recording had median values of 0.67 for diagnoses, 0.93 for surgical procedures and 0.75 over all conditions examined. Practices varied both in the completeness of recording of each condition and in their overall performance. The predictive value (accuracy) of morbidity data was uniformly high for all conditions examined (median 1.00). For repeat prescription drugs, recording on GPASS was both complete and accurate. CONCLUSION: The recording of morbidity data on GPASS for 45-64-year-old patients in a selected group of 41 highly-computerized practices is about 75% complete and highly accurate. For national morbidity studies, it seems likely that amalgamated data from the best GPASS practices will be as complete and accurate as the morbidity statistics currently derived from hospital based activities in Scotland. PMID- 8731628 TI - The importance of diet and physical activity in the treatment of conditions managed in general practice. AB - Evidence from meta-analyses, physiological data and individual studies suggests that diet and exercise are important in the aetiology and treatment of many of the conditions that are managed predominantly in primary care (hypercholesterolaemia, hypertension, diabetes, obesity and excess alcohol intake). However, much of the evidence comes from outside primary care, and it is doubtful whether those studies done in primary care used optimal intervention strategies. A priority for future research should be to demonstrate the feasibility, efficacy and efficiency of lifestyle interventions in a general practice setting. PMID- 8731629 TI - Diagnosis of asthma and chronic obstructive pulmonary disease in general practice. AB - There may be an overlap between the clinical pictures of asthma and chronic obstructive pulmonary disease which hampers a clear distinction between the two diseases. Most symptoms presented by patients do not clearly belong exclusively to either asthma or chronic obstructive pulmonary disease. By the nature of their discipline and training, general practitioners focus mainly on symptoms presented, which do not give a decisive answer in the differential diagnosis between the two diseases. Therefore, general practitioners must rely on objective parameters, such as determining the presence and degree of reversibility of airway obstruction, diurnal peak flow variability, bronchial hyper-responsiveness and allergy. This paper puts forward a pragmatic, primary care definition of asthma and chronic obstructive pulmonary disease. PMID- 8731630 TI - Efficient inhaler devices. PMID- 8731631 TI - Inhaler devices for patients with asthma. PMID- 8731632 TI - Management of opiate dependence. PMID- 8731633 TI - Problem drug users. PMID- 8731634 TI - Patient choice of general practice. PMID- 8731635 TI - Computer medical records. PMID- 8731636 TI - Health promotion. PMID- 8731637 TI - Anecdotes and empiricism. PMID- 8731638 TI - Tobacco regulation--does tax income come before health? PMID- 8731639 TI - Legionnaires' disease as a hospital-acquired infection. PMID- 8731640 TI - A quality assessment scheme for blood glucose monitoring: competition improves performance. AB - Quality assurance procedures are often ignored or forgotten about by wards involved in the blood glucose monitoring of patients. We have devised a complete quality assurance scheme for checking the ward performance of 38 B-M Reflolux meters and two B-M Accutrend meters. Quality control (QC) material, supplied commercially and distributed by the laboratory, is analysed and recorded daily by the wards (internal QC). In addition, a 'spiked', whole blood sample is prepared by the laboratory and distributed for analysis to all wards at six-monthly intervals (external QC). The laboratory also conducts a monthly survey of meter operation (quality assessment) based on a 'scoring system' that allows easy identification of poor performers. Poor performers are counselled accordingly and, if poor performance is persistent, their meter is removed. In our experience, the publication and distribution of ward quality assessment scores has increased competition between wards in terms of avoiding low scores, dramatically improved performance, and ultimately led to improved patient care. PMID- 8731641 TI - Sodium picosulphate compared with polyethylene glycol solution for large bowel lavage: a prospective randomised trial. AB - Mechanical efficiency, patient acceptability and minimal biochemical perturbation are important factors in the choice of lavage solution for bowel preparation. This prospective randomised trial of 59 patients compared a polyethylene glycol bowel lavage solution (Klean Prep) to a sodium picosulphate solution (Picolax) considering their efficacy from both colonoscopists' and radiologists' points of view, patient acceptibility, and plasma biochemical changes. The sodium picosulphate solution was more acceptable to patients than the polyethylene glycol, and resulted in significantly less nausea and vomiting (p = 0.0025), and far fewer consumption difficulties (p < 0.0001). Mean plasma biochemical changes were significantly different for magnesium (p < 0.001), chloride (p < 0.01) and potassium (p < 0.02). Because neither lavage solution displayed a marked advantage for the colonoscopist or radiologist, and sodium picosulphate was more acceptable to patients, sodium picosulphate is the preferred solution for bowel preparation. The changes in plasma biochemistry values are unlikely to cause clinical problems. PMID- 8731642 TI - Incorrect diagnosis of thyroid disease by junior doctors. AB - Thyroid function tests (TFTs) are commonly performed on elderly patients admitted to hospital. Several diseases and commonly used drugs may alter thyroid function test results. We assessed the knowledge of such interactions among 50 junior doctors in our departments: 60% were unaware that (in our laboratory) the normal range for TFTs is the same in those above 65 years as in those below; 72% correctly noted the effect of an acute illness, while only 48% correctly noted the effect of a chronic illness on test results; 72% were well aware of common drug effects such as those of the oral contraceptive pill, but only 22% and 42% knew of the potentially more important interactions in the elderly with the salicylates and phenytoin respectively. Thyroid screening in elderly people is important. The knowledge of interactions between drugs, disease states and the normal range of biochemical assays used in routine thyroid screening needs to be improved if thyroid function tests are to be interpreted appropriately. PMID- 8731643 TI - An audit of 319 deaths across four general practices. AB - Death is a major outcome measure, and the value of an audit of the preventable factors leading to individual deaths is accepted in secondary care. Although approximately 20 of a typical general practitioner's patients will die every year, audits of deaths in general practice are seldom published. This is probably partly due to the difficulties posed by comparability, partly due to the wide range of causes of such deaths and possible contributory factors, and partly due to confidentiality considerations. We describe an audit of all death occurring in our practices over a 12-month period. Although cigarette smoking was considered to exceed all other contributory causes of death, preventable factors occurred in both primary and secondary care sufficiently often for important conclusions to be drawn. Comparison of causes of death between general practices is possible, and such enquiry can demonstrate important quality issues for both primary and secondary care. PMID- 8731644 TI - Influence of teaching on the triage of multiple injury patients. AB - Attempts to improve the standard of early trauma management through the training and instruction of medical personnel have had variable results. Knowledge of trauma care is inadequate among all grades of medical and paramedical staff, and not much improvement has been made by the availability of instruction courses in trauma management. Staff most likely to attend such courses are seemingly already more proficient than staff who choose not to. To achieve maximum benefit from future courses, therefore, we believe attendance should be compulsory for all personnel involved in acute trauma care. PMID- 8731645 TI - Recent advances in the medical management of prostate cancer. AB - Prostate cancer is a 20th century seedling which, because of its attendant morbidity and mortality and the increased longevity of the population, is set to germinate into a substantial economic burden in the next millennium. Most patients with prostatic cancer present with either locally advanced or metastatic disease, for which palliative endocrine therapies are the first-line treatment. The increasingly sophisticated and selective hormonal methods available today, such as the longer-acting formulations of luteinizing hormone-releasing hormone (LH-RH) analogues and newer, better-tolerated, once-daily, non-steroidal anti androgens, have increased the therapeutic options and improved patient quality of life. Maximum androgen blockade, combining medical or surgical castration with an anti-androgen, is an increasingly accepted therapy, and offers the greatest efficacy, particularly for patients with a lesser disease burden. The development of hormone-refractory tumours is still a problem in advanced prostate cancer, although elucidation of the mechanisms involved should offer many potentially fruitful avenues for new therapies. PMID- 8731646 TI - Management of substance misusers on the general hospital ward. AB - Since a high prevalence of physical morbidity is associated with substance misuse, drug misusers are frequently admitted to general hospitals. Medical staff often perceive drug misusers as a particularly difficult patient group, feeling that they lack the knowledge and skill to deal with them adequately without specialist support. We review the common physical complications of substance misuse likely to be encountered in a general hospital, and provide comprehensive guidelines for the assessment and management of drug dependence in this setting. PMID- 8731647 TI - Allergen avoidance in the primary prevention of atopy. AB - Atopy development cannot yet be predicted with certainty, and primary prophylaxis has to be directed at every 'at-risk' infant. However, substantial progress has been made in identifying environmental factors associated with allergic disorders. The process of sensitisation to common allergens may begin in utero. The influence of adjuvants, especially passive smoking, is considerable in individuals with established atopic disease. This article examines the role of intervention during pregnancy and the first year of life in the primary prevention of atopy and suggests a number of measures based on the current evidence. PMID- 8731648 TI - Practical management of primary varicose veins. AB - Varicose veins cause a great deal of morbidity in our population today. Despite the large amount of surgical time spent dealing with the problem, there is still a disappointingly high recurrence rate and many patients are investigated inadequately before surgery. This review considers the assessment of 'simple' varicose veins using a combination of tourniquet tests and a hand-held doppler probe. The place of more sophisticated investigative techniques is also discussed, in particular the value of duplex assessment in localising the variably sited sapheno-popliteal junction. Routine stripping of the long saphenous vein to the below-knee level is likely to decrease the recurrence rate of simple, long, saphenous varicose veins. PMID- 8731649 TI - Legionnaires' disease--a potential hazard of rationalisation. AB - The cases of two elderly patients who complained of general malaise after they had undergone ward transfer (due to rationalisation of the rehabilitation service) are discussed. Both were pyrexial and had signs of pneumonic consolidation. Subsequent investigation revealed that their symptoms were due to infection with Legionella pneumophila. Their cases highlight the problems associated with ward closures and the hazards of stagnant water. PMID- 8731650 TI - Caustic soda burns to the extremities: difficulties in management. AB - Alkali burns to the extremities are unusual, but they are also a cause of significant morbidity, often because of their late presentation. Their management centres around water lavage, burn debridement and split-skin grafting; there is controversy over the timing of surgery. Three unusual cases of caustic soda burn in adults, and the difficulties encountered in their management, are described, and the literature is reviewed. PMID- 8731651 TI - Juvenile dermatomyositis associated with partial lipodystrophy. AB - The case of a child who developed partial lipodystrophy and metabolic abnormalities during treatment of juvenile dermatomyositis is described and discussed. PMID- 8731652 TI - Obstetric factitious disorder: is it more common than we think? AB - This case discusses the diagnosis of obstetric factitious disorder in a 27-year old woman with bleeding per vagina. The case illustrates the need for a high index of suspicion in patients with unexplained physical symptoms during pregnancy, and highlights issues related to subsequent child care. PMID- 8731653 TI - Intracerebral haemorrhage in a young adult: consider amphetamine abuse. AB - We present the case of a young female who suffered a massive intracerebral bleed following the ingestion of a small quantity of amphetamine (speed). Physicians should be aware that amphetamine abuse can lead to cerebrovascular events in young adults. PMID- 8731654 TI - Melioidosis: a rare but not forgotten cause of fever of unknown origin. AB - Melioidosis is widely prevalent in Southeast Asia and northern Australia. Although it is believed to pose no current threat to the populations of developed countries, the increased mobility of people around the world and of Southeast Asian refugees to Western countries may change this. Its long incubation period may put a relatively large number of currently asymptomatic people at risk. It should be considered in the differential diagnosis of any febrile illness in a person who has visited an endemic area, especially if the presenting features are those of fulminant respiratory failure, if multiple pustular or necrotic skin or subcutaneous lesions develop, or if there is a radiologic pattern of tuberculosis from which tubercle bacilli cannot be demonstrated. PMID- 8731655 TI - Benign gastrocolic fistula associated with peritonitis. AB - Gastrocolic fistula is most often related to malignancy or previous gastric surgery. It is an uncommon complication of benign gastric ulceration in patients who have not had a previous operation. Benign gastrocolic fistula associated with peritonitis is extremely rare--this case is only the fourth ever reported. The patient presented with an acute abdomen, and subsequent investigations demonstrated a gastrocolic fistula of benign aetiology. PMID- 8731656 TI - A brief report on malignant triton tumour of the brachial plexus. PMID- 8731657 TI - Epidemiology and prognosis of subungual melanoma in 34 Japanese patients. AB - The incidence of malignant melanoma is much lower in the Japanese than in caucasians. However, amongst the various types of malignant melanoma, the subungual and periungual sites are commonly found in the Japanese. One hundred and fifty-one cases of cutaneous malignant melanoma were seen over a 25-year period at our hospital. We found that, in 34 patients (23%), the subungual region was involved, a high frequency for one institution. We have analysed these patients and looked at their treatment. The finger nails were affected in 21 cases (62%) and the toe nails in 13 cases (38%). The thumb nails or great toe nails were affected in 25 of the 34 patients (73%). In 25 patients, histopathological features of acral lentiginous melanoma were found, with four cases of superficial spreading melanoma and five of nodular amelanotic melanoma. Of the latter group, four mimicked fibrous histiocytic tumour, and one was a desmoplastic malignant melanoma. The proportion of patients presenting with stage III disease decreased after 1982, with a corresponding increase in patients whose tumour thickness was less than 4 mm (stage II). Concurrently, the prognosis for subungual malignant melanoma improved. The 5-year survival rate in each of the periods 1969-82 and 1983-93 was 53 and 87%, respectively. This is similar to that found in plantar malignant melanoma and is felt to be due to a greater public awareness of the condition and to the introduction of effective chemotherapy (the DTIC-AC nitrosurea-vincristine (DAV) regimen). Although the frequency of malignant melanoma is rather low in the Japanese, our data indicate that there is a high incidence of subungual malignant melanoma. Public awareness of the early stage of malignant melanoma seems to have improved prognosis. PMID- 8731658 TI - CD69 expression and tumour necrosis factor-alpha immunoreactivity in the inflammatory cell infiltrate of halo naevi. AB - It has been suggested that the involution of the pigmented lesions of halo naevus (HN) is mediated by an immune response, with the involvement of specific cytotoxic T lymphocytes. To explore further the pathogenesis of HN, skin biopsies from six patients with this condition were obtained and the characteristics of the infiltrating inflammatory cells were studied by immunostaining techniques. We found that the cell infiltrate of HN is mainly composed by CD8+ T lymphocytes that express the activation molecule CD69. Tumour necrosis factor-alpha (TNF alpha) immunoreactivity was detected on the inflammatory cells, a finding that suggests that the infiltrating T cells of HN are actively synthesizing this cytokine. Our results indicate that the infiltrating cells of HN predominantly have an activated cytotoxic phenotype, and suggest that these cells are indeed involved in the regression of the naevomelanocytic naevus of HN. PMID- 8731659 TI - Incidence of familial dermatitis herpetiformis. AB - Dermatitis herpetiformis (DH) and coeliac disease (CD) are gluten-sensitive diseases which have a common immunogenetic background, with the histocompatibility locus antigen (HLA) alleles DQ A1*0501 and B1*0201 in the short arm of chromosome 6. CD is well known to cluster in families whereas DH has not been generally regarded as a familial disease. To study the familial incidence of DH, a prospective study was started in 1969 at the Department of Dermatology of Helsinki University Hospital and in 1976 at the Department of Dermatology of Tampere University Hospital. A total of 1018 patients with DH were diagnosed and questioned for positive family histories. Of the 999 unrelated DH patients, 105 (10.5%) had one or several affected first-degree relatives. The disease in the relatives was either DH (4.4%) or CD (6.1%). Analysis of the 105 families showed that 13.6% of parents, 18.7% of siblings and 14.0% of children were affected, a segregation pattern which fits well to a dominant mode of Mendelian inheritance. Gender may also be important because the first-degree relatives affected with DH were more often females and those affected with CD twice as often females as males. In contrast, there were more males among the 105 propositi with DH and also among the 894 DH patients with no affected relatives. The present study clearly shows that DH is a familial disease in which the first degree relatives can be affected both with DH and CD, presumably because of a common genetic background. The environmental factors which could cause the rather high penetrance of DH and CD in the first-degree relatives of DH patients remain unknown. PMID- 8731660 TI - Calcitonin gene-related peptide, endothelin-1, the cutaneous microvasculature and Raynaud's phenomenon. AB - It has been argued that the digital cutaneous microvasculature is the site of the anomaly which causes Raynaud's phenomenon (RP). Both endothelin-1 (ET-1), a potent vasoconstrictor peptide present in the digital cutaneous microvasculature, and calcitonin gene-related peptide (CGRP), a powerful vasodilator present in digital cutaneous perivascular nerves, have been implicated in the pathogenesis of RP. Circulating ET-1 levels are raised, and there is a diminution of CGRP containing perivascular nerves in finger skin in RP. We undertook a pharmacological study to investigate the sensitivity of the digital cutaneous microvasculature to intradermal ET-1 and CGRP. Differences were found in RP compared with normal digital skin, supporting the idea that the digital cutaneous microvasculature is actively involved in the pathogenesis of RP. In RP, the erythematous response to ET-1 was diminished at both 20 and 5 degrees C (a low temperature at which RP classically occurs) providing pharmacological support for the morphological evidence that in RP there is a deficiency of CGRP-containing nerves in the distal digital skin. PMID- 8731661 TI - Antineutrophil cytoplasmic autoantibodies in patients with systemic sclerosis. AB - Perinuclear type of antineutrophil cytoplasmic antibodies (p-ANCA) have been found in patients with periarteritis nodosa, Churg-Strauss arteritis, or pauci immune idiopathic crescentic glomerulonephritis. Recently, the association of p ANCA and normotensive renal failure, in patients with systemic sclerosis (SSc), was reported. We have studied the incidence of p-ANCA in patients with SSc and have investigated its relationship to clinical and laboratory findings. Sera from 77 patients with SSc were examined by the indirect immunofluorescence (IIF) test, employing an ethanol-fixed human neutrophil and enzyme-linked immunosorbent assay, using purified myeloperoxidase (MPO) as an antigen (MPO-ELISA). The sera from seven patients (9.1%) were p-ANCA positive, by both IIF and MPO-ELISA. One patient died from systemic necrotizing angiitis but the remaining six patients have shown no symptoms of systemic vasculitis or of renal involvement. There was a tendency for patients positive for p-ANCA (anti-MPO antibody) to also be positive for other autoantibodies, such as anti-Sc1-70 antibody, anti-centromere antibody, anti-microsome antibody, anti-thyroglobulin antibody and rheumatoid factor. Although the incidence of p-ANCA (anti-MPO antibody) is low in patients with SSc, and its clinical significance in SSc needs further investigations, this could be a serological marker for certain symptoms in SSc. PMID- 8731662 TI - Plasminogen activator inhibitor type-2 in the lesional epidermis of lupus erythematosus. AB - Under certain pathophysiological conditions epidermal keratinocytes produce urokinase-type plasminogen activator (uPA) or tissue-type PA (tPA). These PAs are subject to regulation by PA inhibitors (PAI), including PAI type-2 (PAI-2). In the normal epidermis, PAI-2 is present in the differentiating suprabasal layers, albeit in the apparent absence of PAs. It has, therefore, been suggested that PAI 2 plays a role in epidermal differentiation not linked to its ability to inhibit PAs. In line with this hypothesis, we have studied, by immunohistochemistry, the distribution of PAI-2, uPA and tPA in the normal and in the lesional epidermis of patients with lupus erythematosus (LE), a disease in which epidermal differentiation is disturbed. The PAI-2 antigen was detectable in the normal epidermis and in the lesional epidermis of LE. In the normal epidermis, the PAI-2 antigen was most pronounced in the granular layer. In the hyperkeratotic epidermal lesions of LE, the PAI-2 antigen was increased. In normal and lesional skin, PAI-2 was distributed along the cell periphery, indicating its association with the cornified envelope. Neither uPA nor tPA was detectable in normal or lesional epidermis. Our findings show that PAI-2 is a major type of PAI in normal epidermis and in the lesional epidermis of LE, and that increased epidermal PAI-2 is observed in a disease which is not associated with an increase in epidermal PAs. The data support the hypothesis that epidermal PAI-2 may have other functions than the regulation of PA activity. PMID- 8731663 TI - The relevance and effect of amalgam replacement in subjects with oral lichenoid reactions. AB - In this study we examined the prevalence of mercury hypersensitivity in patients with oral lichenoid reactions (OLR) and the effect of amalgam replacement in subjects with amalgams adjacent to OLR irrespective of their mercury sensitivity status. One hundred and ninety-seven patients with oral problems were examined: 109 with OLR, 22 with oral and generalized lichen planus, and 66 with other oral diagnoses, including aphthous ulcers and orofacial granulomatosis. Nineteen per cent of patients with OLR reacted to mercury on patch testing, significantly more than in those with generalized lichen planus (0%) and in those with other oral diagnoses (3%). Twenty-two patients with OLR and adjacent amalgams had amalgam replacement and, in 16 of 17 mercury-positive subjects and three of four mercury negative subjects, the OLR resolved after amalgam removal. In conclusion, we found a significantly increased prevalence of mercury hypersensitivity in patients with localized OLR in comparison to subjects with other oral problems. Amalgam replacement resulted in resolution of OLR in the majority of patients with amalgams adjacent to OLR irrespective of their mercury sensitivity status. PMID- 8731664 TI - Differential irritant skin responses to topical retinoic acid and sodium lauryl sulphate: alone and in crossover design. AB - Topically applied all-trans retinoic acid (RA) is often associated with skin irritation. A detailed quantification of RA-induced functional changes in stratum corneum is, however, still limited. Using non-invasive bioengineering techniques of measurements of transepidermal water loss (TEWL), stratum corneum hydration and cutaneous blood flow (CBF), we quantified the irritant effects of 0.05% and 0.1% RA in ethanol on normal skin compared with 1% sodium lauryl sulphate (SLS) in water as a model irritant in a 24-h occlusive patch-test assay. Additionally, in order to document data possibly related to the mechanism of action, skin responses to both compounds applied in tandem was also investigated over 18 days. The extent of the irritant response to 0.05 and 0.1% RA, respectively, were similar, implying analogous irritation potency. While RA caused more intense scaling than SLS, other skin responses to RA were significantly weaker than those due to SLS. An increase in TEWL, on day 7, in RA-exposed sites indicates a secondary delayed impairment of the stratum corneum (SC) barrier. In a tandem design assay, pretreatment with RA appeared to reduce the irritant effects of SLS on SC hydration and CBF. In contrast, pre-exposure to SLS showed a synergestic response in erythema, scaling and TEWL. Our results demonstrate that RA, like SLS, is capable of impairing SC water barrier function, which may be responsible, in part, for the irritation associated with its topical use. However, the distinctive biological responses to these compounds suggest a different mode of action of RA and SLS. In addition, the precise reason for the unique results observed in the tandem-design assays is not clear. PMID- 8731665 TI - Topical retinoic acid changes the epidermal cell surface glycosylation pattern towards that of a mucosal epithelium. AB - Topical all-trans retinoic acid (RA) produces a number of epidermal changes which are indistinguishable from those observed following treatment with a local irritant, namely sodium lauryl sulphate (SLS). This observation has led to criticism that the efficacy of RA in disorders such as photoageing, is merely a result of irritancy. In stratified epithelia, the cellular differentiation process is characterized by a stepwise synthesis of cell surface carbohydrates, and each type of stratified epithelium has its own specific pattern of carbohydrate expression. Glycosyltransferases, which are responsible for carbohydrate synthesis, are influenced by retinoids. Thus, we investigated whether epidermal cell surface glycosylation is altered in skin treated with topical RA, and contrasted it with changes induced by topical SLS. Skin biopsies were obtained from seven normal volunteers who had been treated, on three separate areas of buttock skin, with single applications of 0.1% RA, 2% SLS, or vehicle creams, followed by 4-day occlusion. Biopsies were assessed immunohistologically using highly specific monoclonal antibodies to cell surface carbohydrates (types 1, 2 and 3 chain structures), previously demonstrated in the epidermis and in oral mucosal epithelium. Although type 1 chain structures were not demonstrated in any of the samples, the distribution of type 2 and 3 chain structures in RA-treated epidermis was altered towards that seen in a mucosal epithelium. T antigen, a mucin-type cell surface carbohydrate structure normally expressed throughout the epidermis, was only observed in the granular layer of RA treated epidermis--a feature of mucosal epithelia. Ley, normally only seen in non keratinized buccal epithelium, was strongly expressed in RA-treated epidermis. In contrast, the glycosylation pattern of the SLS-treated epidermis was not significantly different from that observed after vehicle treatment. Thus, RA treatment converts normal stratified epithelium towards the phenotype of mucosal epithelium with a decrease in T antigen and a concomitant increase in Ley. These changes are not observed following treatment with SLS and identify an important difference between RA effects and irritancy. PMID- 8731666 TI - A comparison of the culture and growth of dermal papilla cells from hair follicles from non-balding and balding (androgenetic alopecia) scalp. AB - Male pattern baldness is a common, androgen-dependent skin problem in adult men which is not well understood, although androgens are believed to act on the hair follicle via the mesenchyme-derived dermal papilla situated in the middle of the hair follicle bulb. Since dermal papilla cells retain specific characteristics in culture, such as hair-growth promoting ability and appropriate features of the mechanism of androgen action, dermal papilla cells from follicles undergoing androgen-stimulated miniaturization may provide a useful in vitro model system. Therefore, dermal papilla cells have been derived from intermediate follicles from balding and nearly clinically normal sites of men with androgenetic alopecia. Balding dermal papillae were much smaller than non-balding ones and grew much less well under normal growth conditions. Supplementing the medium with human serum, rather than fetal calf serum, increased both the yield of established cultures and the number and health of the dermal papilla cells produced. Non-balding cells also grew faster in human serum. Balding cells retained the normal fibroblastic shape and aggregative behaviour of dermal papilla cells, but always grew less well than non-balding cells. Nearly clinically normal dermal papillae were similar, or slightly smaller, in size to non-balding ones, but their growth resembled balding cells. Since balding dermal papilla cells can be cultured, though with much greater difficulty than nonbalding ones, and exhibit differing growth characteristics to non-balding cells, they merit further investigation which may increase our understanding of, and ability to control, androgenetic alopecia. PMID- 8731667 TI - Interaction of cultured human keratinocytes with liposomes encapsulating silver sulphadiazine: proof of the uptake of intact vesicles. AB - There is evidence to suggest that human keratinocytes grown in vitro are capable of engulfing and subsequently disintegrating intact liposomes. However, as the liposomes used in this context did not carry an electron-dense marker, the possibility that the lamellar structures seen within the keratinocytes were composed of material produced within the cell could not be excluded. We therefore decided to investigate liposome-keratinocyte interaction using an electron-dense marker. Human keratinocytes obtained from juvenile foreskins were cultured in a serum-free medium, and subconfluent cultures were exposed to liposomally encapsulated and free silver sulphadiazine 1% (SSD), and a corresponding vehicle, for 5 min to 24 h. After fixation ultra-thin sections were analysed electron microscopically at magnifications of up to x85,000. Many keratinocytes treated with liposomal and free SSD showed marked damage to the plasma membranes and the cell organelles. The phagocytosis of intact liposomes was demonstrated by the appearance of silver-labelled unilamellar vesicles within the cytoplasm of undamaged keratinocytes. The labelled liposomes were found enclosed in cellular unit membranes, i.e. in lysosomes. In addition, perinuclear disintegration and release of the entrapped marker were observed. Silver particles, as present in liposomally encapsulated SSD, were found to be adequate markers for electron microscopy. Our results confirm the phagocytosis of intact liposomes by keratinocytes in vitro. In addition, the cytotoxic effects of liposomal (intended for the treatment of burns) and free SSD on human keratinocytes were studied in detail. Many keratinocytes treated for 10 min or more were severely affected. PMID- 8731668 TI - Carcinoembryonic antigen and related antigens expressed on keratinocytes in inflammatory dermatoses. AB - Carcinoembryonic antigen (CEA) is recognized as being important not only as a tumour marker, but also as a cell adhesion molecule in the metastasis of malignant tumour cells, and in the development of inflammatory disorders in some organs. However, the role played by this antigen in inflammatory dermatoses has not been studied previously. To investigate the expression of CEA in inflamed skin, we performed immunohistochemical studies to detect the antigen in both frozen and formalin-fixed, paraffin-embedded biopsy specimens of inflammatory dermatoses, using five different monoclonal antibodies and one polyclonal antibody against human CEA or CEA-related molecules. CEA and CEA-related antigens were expressed by epidermal keratinocytes and by some infiltrating inflammatory cells in inflamed skin. The expression was observed primarily in the upper part of the malpighian layers of the epidermis. On the basis of the distribution of this expression, we suggest an additional role for CEA-related molecules, differing from their roles as tumour markers and adhesion molecules, namely that these molecules may be associated with the state of differentiation of epidermal keratinocytes. Unexpected expression of CEA-related antigens was seen on normal follicular keratinocytes. Thus, it is conceivable that CEA-related antigen may also play a role in the biology of hair follicles. PMID- 8731669 TI - The role of two endogenous proteases of the stratum corneum in degradation of desmoglein-1 and their reduced activity in the skin of ichthyotic patients. AB - We investigated the role of stratum corneum (SC) trypsin-like and chymotrypsin like serine proteases in the degradation of desmoglein-1 (DSG-1) in the SC sheet. DSG-1, whose presence in the SC sheet was confirmed by Western blot analysis, was degraded completely during incubation of the SC sheet in Tris buffer. The degradation of DSG-1 was inhibited by the addition of protease inhibitors, such as aprotinin or a mixture of leupeptin and chymostatin. Either leupeptin or chymostatin alone did not inhibit its degradation. These results indicated that both trypsin-like and chymotrypsin-like serine proteases are involved in the degradation of DSG-1. We further examined the activities of the two proteases in the SC obtained from patients with ichthyosis vulgaris, in whom SC desquamation is abnormal. The enzymatic activities measured using synthetic substrates were significantly decreased in these ichthyotic SC samples. This result supports the idea that these proteases play an important role in normal SC desquamation. PMID- 8731670 TI - Expression of CD44 isoforms in basal cell carcinomas. AB - The expression of CD44 isoforms (CD44std, CD44v6, CD44v10) was investigated by an immunohistochemical technique in 42 basal cell carcinomas (BCC) of the superficial and nodular variety. All BCCs studied displayed very low amounts of CD44std, a receptor for hyaluronic acid. Except for single CD44std-positive cells located preferentially in the central parts of the BCC nests, the bulk of the tumour formations were CD44std-negative. CD44v6 showed a heterogeneous distribution pattern accentuated in the peripheral palisading tumour cells. In superficial BCCs, the labelling intensity for CD44v6 increased with the size of the tumour nests. CD44v10 was not detectable in BCCs. Our findings support the notion that CD44v6 is not linked to the metastatic proclivity of tumours originating from keratinocytes. We suggest that the very low expression of the receptor for hyaluronic acid (CD44std) may be one of the factors which block the formation of metastases from BCCs. PMID- 8731671 TI - Pachydermatous eosinophilic dermatitis. AB - We report three South African black teenage girls with extensive pruritic papular lesions arising on a pachydermatous base, resembling severe atopic dermatitis or onchodermatitis. All three had peculiar hypertrophic genital lesions and peripheral blood eosinophilia. Histological studies showed an eosinophil-rich lymphohistiocytic infiltrate and variable fibrosis. Extensive fibrillar extracellular deposition of eosinophil granule major basic protein was demonstrated by an indirect immunofluorescence technique. A beneficial therapeutic effect was obtained using dapsone, prednisolone and cetirizine. The term pachydermatous eosinophilic dermatitis is proposed and its position among other conditions characterized by peripheral blood and skin tissue eosinophilia, is discussed. PMID- 8731672 TI - Side-effects and complications of flashlamp-pumped pulsed dye laser therapy of port-wine stains. A prospective study. AB - The flashlamp-pumped pulsed dye laser (FPDL) was the first laser system specifically developed for the treatment of cutaneous vascular lesions such as port-wine stains (PWS), telangiectases and haemangiomas. Its theoretical advantages have been verified by numerous excellent clinical results. As there are few systematic studies on the adverse effects of this laser system, we investigated 100 consecutive patients who received FPDL therapy of PWS. Pain during and after laser therapy was a common complaint. Post-treatment erythema occurred in 29%, oedema in 73%, formation of bullae in 1%, and serous crusting in 46-83% of patients (based on our observations and the patients histories, respectively), all of which lasted for a maximum of 7 days. Furthermore, complications such as impetigo-like crusting occurred in 25%, bleeding in 12%, a pyogenic granuloma in 1%, hyperpigmentation in 27%, hypopigmentation in 1%, atrophic scarring in 3%, and an hypertrophic scar in 1%. Although most of these changes were only focal, and were predominantly transient, our results indicate that the incidence of side-effects and complications produced by the FPDL (wavelength 585 nm, pulsewidth 450 microseconds) may be higher than previously documented in the literature. PMID- 8731673 TI - The value of dynamic hepatic scintigraphy and serum aminoterminal propeptide of type III procollagen for early detection of methotrexate-induced hepatic damage in psoriasis patients. AB - Oral methotrexate (MTX) is a highly effective drug for the treatment of severe psoriasis. A limitation of this treatment is its potential hepatotoxicity. In the present prospective study the value of dynamic hepatic scintigraphy (DHS) and serum aminoterminal propeptide of type III procollagen (PIIINP) were investigated as screening methods for early detection of MTX-induced hepatic damage. These relatively non-invasive procedures were compared with the liver biopsy classification, until now the gold standard to assess MTX-induced liver damage. Twenty-five MTX patients were studied. The mean cumulative MTX dose was 3.9 g (range 0.2-11.1 g). Twenty-one patients had a normal liver histology (grade I), three patients had steatosis (grade II), and one patient mild fibrosis (grade IIIA). Seven additional patients with non-MTX related hepatic cirrhosis were included as disease controls. DHS showed a clear-cut separation between the portal contribution of the MTX patients with grade I liver histology, and that of all other patients. A portal contribution larger than 52% was associated with a > 95% chance of normal liver histology. If this cut-off value had been used to postpone the liver biopsy, this would have resulted in at least a 55% reduction in the number of biopsies in patients with a normal liver histology. DHS appeared to be very promising as a screening test to differentiate between the presence or absence of MTX-induced hepatic damage, but appeared not suitable to grade the severity of hepatic damage. Although a global relationship was demonstrated between serum PIIINP concentration and hepatic damage, single measurements in individual patients were not reliable. The combination of PIIINP measurements with DHS had only a limited additional value above DHS alone. The present study indicates that DHS has great promise for the detection of early MTX-induced hepatic damage. Pending further studies, regular liver biopsies remain mandatory for the safe prolonged use of MTX in psoriasis patients. PMID- 8731674 TI - Detection of EBER nuclear RNA in T-cell lymphomas involving the skin--an in situ hybridization study. AB - To examine the relationship of Epstein-Barr virus (EBV) to cutaneous T-cell lymphoma (CTCL), a retrospective study was performed using formalin-fixed paraffin-embedded skin biopsies from 12 patients with cutaneous T-cell lymphoma. All cases were immunophenotyped with a panel of monoclonal antibodies including CD45RO, CD43, MB-2, CD20, and CD68. In situ hybridization studies to detect the EBV genome were performed, using the EBV-RNA probes for the early RNAs of EBV, namely EBER and BHLF. Among the 12 patients, six had primary CTCL, and six had secondary CTCL. The nasal cavity was the most frequent extracutaneous primary site. Immunophenotypically, all of the cases were of the T-cell lineage. In situ hybridization demonstrated a diffuse strong positive reaction for EBV transcripts (EBER) nuclear RNA in five out of 12 cases (41.7%) and a negative reaction for BHLF nuclear RNA probe in all the cases studied. Angiocentric lymphomas were positive in three out of the five cases (60%) studied. Secondary CTCL had a higher correlation with EBV than did primary CTCL, especially primary nasal T cell lymphoma with subsequent cutaneous involvement. The EBV genome in lymphoma cells existed in a latently infected form but not in an actively replicating form. Three out of the five EBV-positive patients died, 8 months to 4 years after diagnosis. EBV infection alone in the lymphoma does not seem to affect the prognosis of CTCL patients significantly. PMID- 8731675 TI - Merkel cells participate in the induction and alignment of epidermal ends of arrector pili muscles of human fetal skin. AB - The follicular proximal ends of arrector pili muscles anchor the bulge areas of human hair follicles. However, the location of the ends of these muscles towards the epidermis, especially the fine points of attachment, is not fully understood. Conventional serial sections of the scalp skin of a 12-month-old infant were examined. Most of the distal ends were situated in the upper dermis, while some seemed to be in close contact with the epidermal basal layers. In vertical sections of the scalp skin of 36-week-old fetus, double immunoenzyme staining showed a close topographic correlation between the epidermal ends of the arrector pili muscles and the epidermal and dermal Merkel cells. These findings allow speculation that fetal Merkel cells participate in the induction and alignment of arrector pili muscles. PMID- 8731676 TI - HIV-associated eosinophilic folliculitis: case report and review of the literature. AB - We report human immunodeficiency virus (HIV)-associated eosinophilic folliculitis in a previously well 40-year-old Portuguese lady. This condition, although similar to Ofuji's disease, has distinguishing features which allow the two to be differentiated. To the best of our knowledge, this is the first reported case of HIV-associated eosinophilic folliculitis in a female and in a patient in whom the HIV was heterosexually transmitted. Our patient is also unusual in being, we believe, only the second reported in whom this eruption was the presenting feature of HIV infection. PMID- 8731677 TI - Cutaneous reactions following herpes zoster infections: report of three cases and a review of the literature. AB - Three patients, one healthy and two immunocompromised, developed cutaneous reactions that histologically mimicked granuloma annulare at sites of resolved varicella-zoster virus (VZV) reactivation infections. Variable latency periods between the infection and the granulomatous reaction were noted. As in other case reports, the presence of VZV DNA in these lesions was inconsistently demonstrated by the polymerase chain reaction (PCR) and appears more common in early, as opposed to late, post-zoster granulomas. In addition to various granulomatous reactions, vasculitic and neoplastic eruptions following resolved VZV infections have been described and are reviewed here. Therapeutically, topical, intralesional and systemic corticosteroids, as well as acyclovir, have been tried with inconsistent results. Although the pathogenesis remains unclear, the presence of VZV DNA in early lesions that histologically do not display viral cytopathic changes, suggests the virus induces an atypical delayed hypersensitivity reaction not affected by antiviral therapy. PMID- 8731678 TI - Generalized pruritus in anorexia nervosa. AB - We report a 19-year-old woman with the associated disorders of generalized pruritus, hypertrichosis and anorexia nervosa, all of which had been present for approximately 4 months. Psychotherapy induced both weight gain and resolution of the pruritus. We propose that anorexia-associated pruritus be considered one of the important cutaneous signs in anorexia nervosa. PMID- 8731679 TI - The scleroatrophic syndrome of Huriez: a cancer-prone genodermatosis. AB - We report a 24-year-old woman, her 6-year-old son and her 17-month-old daughter, who all suffer from a rare congenital genodermatosis first delineated by Huriez et al. in the 1960s. The clinical features of this autosomal dominant condition include scleroatrophy of the hands and feet, nail hypoplasia, mild palmoplantar keratoderma and hypohidrosis. Histological changes are non-specific, but immunohistological and ultrastructural examination in our index patient revealed an almost complete absence of epidermal Langerhans cells in the affected skin. This new finding may be linked to the cancer proneness of the scleroatrophic skin. In this family, the grandmother had died at the age of 37 years from metastatic squamous cell carcinoma which had arisen on the thenar eminence. PMID- 8731680 TI - Amelanotic superficial spreading malignant melanoma mimicking Bowen's disease. AB - A 67-year-old woman presented with a scaly, erythematous plaque, for which a clinical diagnosis of Bowen's disease was made. However, incisional biopsy established the diagnosis of amelanotic superficial spreading malignant melanoma. Biopsy thus avoided the use of inappropriate destructive therapy, such as cryotherapy or cautery. This case illustrates the importance of obtaining a histological diagnosis, prior to treatment, in suspected cases of Bowen's disease. PMID- 8731681 TI - Vulval cicatricial pemphigoid may mimic lichen sclerosus. AB - We report two patients with vulval signs suggestive of lichen sclerosus. This diagnosis was never substantiated histologically and microscopic appearances were non-specific. In both cases the development of lesions at other cutaneous and mucosal sites alerted us to the possibility that the autoimmune bullous disease, cicatricial pemphigoid, accounted for the entire clinical picture. Lichen sclerosus and cicatricial pemphigoid are distinctive conditions which clinically are usually readily differentiated on cutaneous surfaces. When the site of presentation is on the vulva the clinical picture may become obscure and this can lead to diagnostic confusion. It is important to establish the correct diagnosis as this predicts the clinical course and prognosis in each case. PMID- 8731682 TI - Indeterminate cell histiocytosis--a clinicopathological entity with features of both X- and non-X histiocytosis. AB - An otherwise healthy 50-year-old woman presented with a 6-month history of having developed more than 100 generalized, non-confluent, reddish-brown, partially yellow-coloured papules. A non-epidemotropic, monomorphous infiltrate of vacuolated mononuclear, and occasionally multinuclear, histiocytes, positive for factor XIIIa and macrophage markers HAM56 and KiM1p, was consistent with the clinical impression of generalized eruptive histiocytomas. However, the additional reactivity for S100 protein, in the absence of features of histiocytosis X, suggested a diagnosis of indeterminate cell histiocytosis (ICH). Further immunohistochemical studies, performed on snap-frozen material, characterized the lesions as being diffusely positive with LN3 (HLA-DR), Leu4 (CD3) and Leu3 (CD4), the infiltrate in the upper dermis as reactive for OKT6 (CD1) and IOT6c (CD1c), and the infiltrate in the lower dermis as reactive for a variety of macrophage markers. Ultrastructural studies showed various non specific features of histocytic disorders, but no Birbeck granules. Our findings confirm those of previous reports suggesting that ICH is a distinct histiocytic entity, characterized by immunophenotypic features of both X- and non-X histiocytoses. Generalized eruptive histiocytoma seems to be an early indeterminate stage of various non-X histiocytic syndromes including ICH, multicentric reticulohistiocytosis, xanthogranuloma and xanthoma disseminatum. The distribution pattern of the various X/non-X histiocytic markers suggests dermal arrest of antigen-presenting cells during their physiological trafficking from the skin to the lymph nodes. PMID- 8731683 TI - Chilblain lupus erythematosus of Hutchinson responding to surgical treatment: a report of two patients with anti-Ro/SS-A antibodies. AB - We report two patients with chilblain lupus erythematosus of Hutchinson (CL) who responded to surgical treatment. One of them was a 72-year-old woman (case 1), and the other a 62-year-old man (case 2). We attempted to treat these patients by excising the lesions and subsequently performing full-thickness free skin grafting, using skin from the abdominal region. No recurrence was seen in the operated area 7 years (case 1) and 3 years (case 2) after surgery. However, lesions persisted in the areas not operated upon, and in the areas where lesions had not been adequately excised. These results suggest that surgical removal of local factors reduces the rash in these cases. In addition, both patients were serologically positive for the anti-Ro/SS-A antibody suggesting that local expression of the Ro/SS-A antigen may be involved in the pathogenesis of the skin lesions. To our knowledge, full thickness free skin grafting has not been used previously to treat CL-associated skin lesions, and is promising as a treatment for patients who do not respond to conventional means. PMID- 8731684 TI - Coexistence of palmoplantar lichen planus and lupus erythematosus with response to treatment using acitretin. AB - Lichen planus and lupus erythematosus may occur as an overlap syndrome. Here we report the clinical characteristics of a 49-year-old man with palmoplantar lichen planus and lupus erythematosus. He showed a remarkable clinical response to treatment with acitretin, which resulted in prolonged remission of the disorder. PMID- 8731685 TI - Microscopic polyangiitis. A systemic vasculitis with a positive P-ANCA. AB - A young girl presented with a purpuric rash on lower limbs, fever, eosinophilia, peripheral neuropathy and progressive renal insufficiency. She developed vesicles, purpuric macules and papules on the head, several nodules on the palmar sides of hands and fingers, splinter haemorrhages, and a disfiguring, facial oedema. A renal biopsy specimen disclosed a focal and segmental necrotizing glomerulonephritis with crescents. Peripheral ANCA with antimyeloperoxidase specificity [P-ANCA (MPO)] was positive and cytoplasmic ANCA with PR3 specificity was negative. Treatment with prednisone and cyclophosphamide was started with a good clinical response, stabilization of renal insufficiency and disappearance of P-ANCA (MPO). Our case fulfils the diagnostic criteria for microscopic polyangiitis (microscopic polyarteritis, MPA), namely a segmental necrotizing and crescentic glomerulonephritis associated with extrarenal vasculitis involving small-sized vessels, without granulomas or asthma. This is a rare disease, which has a poor prognosis in the absence of aggressive therapy, and is infrequently reported in dermatological journals. PMID- 8731686 TI - Cutaneous manifestations of congenital afibrinogenaemia. AB - Skin problems from coagulation disorders have rarely been described in the dermatological literature. Here we report a patient with a complete congenital absence of fibrinogen which led to leg ulceration, necrosis of the toes and a life-threatening haemorrhage following skin biopsy. This patient shows that leg ulcers may have a complex aetiology and can reflect serious underlying disease. PMID- 8731688 TI - Spontaneous regression in angiocentric T-cell lymphoma. AB - An 8-year-old boy presented with a 10-week history of ulcerating lesions which were histologically and immunocytochemically consistent with the diagnosis of angiocentric T-cell lymphoma. The disease was limited to the skin and resolved with no chemotherapy. Angiocentric T-cell lymphoma is commonly a disease with considerable morbidity and is often fatal. Epstein-Barr virus (EBV) could not be identified in involved tissue by immunostaining or by in situ hybridization. We consider whether the uncharacteristic absence of EBV in this case has prognostic significance. PMID- 8731687 TI - Acute myelocytic leukaemia associated with Kaposi's sarcoma in a patient without serum antibodies to human immunodeficiency virus type 1. AB - We report a 35-year-old man with acute myelocytic leukaemia and Kaposi's sarcoma, whose serum was non-reactive for antibodies to human immunodeficiency virus type 1 by enzyme-linked immunosorbent assay, Western blot and immunofluorescence. Complete remission of both the acute leukaemia and the Kaposi's sarcoma followed treatment with mitoxantrone and cytosine arabinoside. We speculate that the rapid regression of the lesions of Kaposi's sarcoma might be related to mitoxantrone and to the return to normal of his peripheral helper and suppressor T-cell counts. PMID- 8731689 TI - Dermatomyositis responding to pentoxifylline. PMID- 8731690 TI - Morphoea following Blaschko's lines. PMID- 8731691 TI - Extensive extrafacial granuloma faciale of the scalp. PMID- 8731692 TI - Persistent scleredema of Buschke in a diabetic: improvement with high-dose penicillin. PMID- 8731693 TI - Bullous pemphigoid and ulcerative colitis: a report of two cases and description of immunoblot findings. PMID- 8731694 TI - Dermatomyositis with linear lesions. PMID- 8731695 TI - Periorbital oedema and erythema in systemic lupus erythematosus. PMID- 8731696 TI - Pemphigoid gestationis with autoantibodies exclusively directed to the 230-kDa bullous pemphigoid antigen (BP230Ag) PMID- 8731697 TI - Follow-up of circulating ICAM-1 in malignant melanoma: correlation with the clinical course of the disease. PMID- 8731698 TI - Leg ulcers and Klinefelter's syndrome: role of PAI-1. PMID- 8731699 TI - Nodular solar degeneration following herpes zoster. PMID- 8731700 TI - Fibrosing colonopathy in children with cystic fibrosis. PMID- 8731701 TI - A reader's guide to the evaluation of causation. AB - In this article, we review the criteria used to establish a causal association. A literature search strategy will be outlined. We present a clinical scenario that was formulated to examine the evidence for nonsteroidal anti-inflammatory drugs as a possible cause of gastrointestinal ulceration and bleeding. Several of the studies identified in the literature search were critically appraised and the criteria needed to confirm a causal link were then applied. These guidelines may be employed to not only demonstrate the aetiology of a disease but have wider applicability in terms of determining whether a medication may be causing certain adverse events. PMID- 8731702 TI - Your child is brain dead. AB - One to two per cent of admissions to Paediatric Intensive Care Units eventually fulfil the criteria for brain death, implying the need for very difficult decisions. Brain death is defined as irreversible loss of function of the whole brain. The diagnostic criteria caused a great deal of anxiety but are now the subject of a consensus approach. When the situation can be anticipated it is of immense value for the professional staff to develop a good working relationship with the parents to help and support them through the phase of impending disaster and facing the issue when the time actually comes. However, it is vital to help parents to make their own decision regarding continuation or otherwise of life support and they should be supported in whichever decision they take. They must be absolutely convinced that the child is brain dead and this territory may have to be covered again and again in discussion, questions must be answered factually, and time allowed for reflection. Stage management of the process of 'switching off' is vital and the parents' wishes may vary widely from one family to another. They must be warned what is likely to happen and provided with appropriate privacy and support for expression of their grief. The question of asking for post-mortem permission has to be handled sensitively and long-term support for the parents must be offered. Education of undergraduate and postgraduate doctors in this area is now receiving more attention with skills being increased by video teaching and role play. It should not be overlooked that the professional staff attending such patients sometimes require counselling and support themselves. PMID- 8731704 TI - Making medical recruitment practices fair. AB - The fact that in the past each medical student was assured of a post when qualified has led many in medicine to believe that selection for recruitment is merely a formality based on qualification and accreditation. Consequently little thought has gone into developing detailed person or post selection criteria. As a result most recruitment panel members are unclear on what basis to discriminate between equally qualified candidates and invariably plump for 'the face that fits'. This practice discriminates against women, black doctors and those white men not quite acceptable in manner, dress or attitudes. In this article, advice is offered on the development of equality strategies for public sector organisations and on the implementation of fair practices. PMID- 8731703 TI - Subclinical thyroid disease. AB - Thyroid disease can roughly be divided into functional and anatomical disorders. Subclinical disease is by definition not accompanied by symptoms or signs and usually goes unrecognized for the bearer (and the observer). In this communication an overview will be given of existing literature and some own results concerning subclinical hypothyroidism, subclinical thyrotoxicosis and thyroid incidentalomas. Apart from definitions, data on prevalence, clinical effects, prognostic significance and the need for and response to therapy will be discussed. PMID- 8731705 TI - Aerosol delivery systems for bronchial asthma. AB - Many different inhalation devices are now available for the treatment of asthma. Pressures towards the elimination of chlorofluorocarbon propellants are driving forward development of new devices-as are obvious commercial pressures, including the increased availability of generic formulations. We still, however, often cannot tell exactly where within the lung we want to target a particular medication, be it a bronchodilator or a steroid. The basic processes of aerosol deposition are readily comprehensible. Nevertheless, even under carefully supervised inhalation conditions, one can only roughly estimate where the medication is deposited. We can, however, hope to give our patients good guidance on how to make the best use of a metered-dose inhaler or a jet nebuliser. From the array of available devices, we will increasingly be able to select the most comfortable and convenient for the patient-and therefore most likely to encourage good compliance. PMID- 8731706 TI - Pulmonary embolism--the role of thrombolytic therapy in its management. AB - In massive pulmonary embolism where there may be evidence of right ventricular dysfunction and acute pulmonary hypertension, anticoagulation therapy alone may prove inadequate. In such situations use of thrombolytic agents produces an improvement in haemodynamics compared to anticoagulants alone, although studies to date have been too small to address the issue of mortality benefit. It would appear that all age groups gain benefit from the use of thrombolytics. Studies that have compared thrombolytic agents and anticoagulants are discussed. In addition, the issues of the choice of thrombolytic agent and the different modes of delivery of therapy are addressed. PMID- 8731707 TI - Do accident and emergency senior house officers know the British guidelines on the management of acute asthma? AB - Avoidable deaths from asthma continue, even in hospital. Since the management of acute severe asthma is often initiated in the Accident and Emergency department, it is crucial that staff there have adequate knowledge. An anonymous questionnaire, containing items based on chart 6 of the UK guidelines, was completed by 66 Accident and Emergency Senior House Officers from the Yorkshire region. The study aim was to establish these doctors' levels of knowledge about the recommended management of acute asthma in Accident and Emergency. The median score was 10 (out of a possible 24) and the interquartile range 8-13. Further efforts are required to implement these guidelines so that the best patient outcomes can be achieved. PMID- 8731708 TI - Intestinal permeability and orocaecal transit time in elderly patients with Parkinson's disease. AB - The aetiology of weight loss in patients with Parkinson's disease is likely to be multifactorial. We studied 15 patients with Parkinson's disease and 15 age- and sex-matched controls and looked for evidence of malabsorption due to small bowel bacterial overgrowth or alteration of intestinal permeability. There was a marked increase in orocaecal transit time in the patients with Parkinson's disease, although lactulose hydrogen breath testing did not show evidence of small bowel bacterial contamination. Intestinal permeability measured by the differential sugar absorption test was also deranged. There was reduced absorption of mannitol in patients with Parkinson's disease while lactulose absorption was similar in both groups, suggesting decreased non-mediated uptake across the enterocyte brush border membrane in patients with Parkinson's disease. PMID- 8731709 TI - Hirsutism in the United Arab Emirates: a hospital study. AB - In a study of 102 women (Arabs and Asians) with hirsutism, polycystic ovary syndrome was diagnosed in 93 patients (91%) including 26 cases with hirsutism and regular menstrual periods. In more than half of the cases the diagnosis was made on the basis of transabdominal ultrasonography, which was the single test with the highest yield. Luteinising hormone hyperresponsiveness to gonadotropin releasing hormone was positive in only seven of 50 patients tested. Late onset congenital adrenal hyperplasia due to 21-hydroxylase deficiency was identified in two cases, thus forming a prevalence of 8% in those who underwent adrenocorticotropin stimulation. In one case, polycystic ovary syndrome was an associated condition. In this study of a population of women with hirsutism and regular menses, the majority had polycystic ovaries. It is suggested that the term 'idiopathic or racial' hirsutism should not be applied unless a thorough evaluation has failed to reveal a diagnosable underlying disorder. PMID- 8731710 TI - Clomiphene-induced pituitary apoplexy in a patient with acromegaly. AB - We describe the case of an acromegalic woman with primary infertility who was treated with clomiphene, and subsequently developed pituitary apoplexy. PMID- 8731711 TI - Ectopic ACTH syndrome: redefinition and case report. AB - A case is reported which supports the principle that ACTH precursors best characterise so-called ectopic ACTH-syndrome, which should therefore more accurately be referred to as ectopic ACTH-precursor syndrome. PMID- 8731712 TI - Iris metastasis: an unusual cause of anterior uveitis. AB - We report a case of anterior uveitis due to iris metastasis in a patient with known carcinomatosis. PMID- 8731713 TI - Late presentation of a solitary metastasis of renal cell carcinoma as an obstructive duodenal mass. AB - We present a rare case of late solitary metastasis of renal cell carcinoma presenting with duodenal obstruction. Metastatectomy resulted in relief of symptoms and may improve long-term survival. PMID- 8731714 TI - Postural hypotension in a patient with acute myelitis. AB - A case of acute transverse myelitis with severe postural hypotension is described. PMID- 8731715 TI - Hearing loss and leukocytosis. PMID- 8731716 TI - Toxic epidermal necrolysis following chlorpromazine ingestion complicated by SIADH. PMID- 8731717 TI - Toxic epidermal necrolysis associated with indomethacin therapy. PMID- 8731718 TI - Neuroleptic malignant syndrome with normal creatine kinase. PMID- 8731719 TI - Flavobacterium meningosepticum meningitis in an adult with acute leukaemia. PMID- 8731720 TI - Eikenella corrodens thumb osteomyelitis. PMID- 8731721 TI - Pressor effect of metoclopramide in phaeochromocytoma. PMID- 8731722 TI - Fibrate monotherapy and profound hypoalphalipoproteinaemia. PMID- 8731723 TI - [The actors of osteoarthritis]. PMID- 8731724 TI - [Mechanisms of degradation of argecanes in osteoarthritic cartilage]. PMID- 8731726 TI - [Measurement of the progress of osteoarthritis]. PMID- 8731725 TI - [Molecular mechanisms implicated in the inhibition of collagenase expression by diacerheine]. PMID- 8731727 TI - [Cartilage and arthroscopy]. PMID- 8731728 TI - [Interleukin-1, cartilage and osteoarthritis]. PMID- 8731729 TI - [Effects of interleukin-1 on human chondrocytes cultured in aggregates]. PMID- 8731730 TI - [Beneficial effects of treatment with diacerhein in guinea pigs with osteoarthritis]. PMID- 8731731 TI - [Anti-inflammatory and analgesic treatment of osteoarthritis]. PMID- 8731732 TI - [Study of ART 50 in daily rheumatological practice]. PMID- 8731733 TI - [Evaluation methods of osteoarthritis: apropos of the ECHODIAH study]. PMID- 8731734 TI - [Classification and clinical presentation of pancreatitis]. AB - All pancreatitis share identical clinical features, which are evocative of a pancreatic disease; the diagnosis will be established by an enhanced level of pancreatic enzymes in the blood. Actually, we have to consider of one hand acute pancreatitis which heal completely when the cause has been treated (whatever it is of pancreatic or of extra-pancreatic origin) and on the other hand chronic pancreatitis characterized by the presence of fibrosis. Among chronic pancreatitis, one distinguishes chronic obstructive pancreatitis, upstream of an obstacle on the main pancreatic duct from chronic calcifying pancreatitis which is characterized by a diffuse, heterogeneous destruction of acinar tissue, replaced by dense fibrosis, the main etiology of which still remains alcohol. PMID- 8731735 TI - [Imaging of pancreatitis]. AB - Acute and chronic pancreatitis involve different problems. In acute pancreatitis, the method of choice is CT scan, which provides a precise description of the lesions as a basis for establishing criteria of severity. Discovery of infection often requires scan-guided puncture. For chronic pancreatitis, a greater range of examinations is available. The CT scan is the best noninvasive method. Ultrasound examination can be repeated for follow-up. Echoendoscopy shows early signs. Excellent diagnostic results are provided by retrograde endoscopic cholangiopancreatography. It is indispensable for the presurgery work-up and sometimes for differential diagnosis of cancer. PMID- 8731736 TI - [Prognosis and intensive care of severe acute pancreatitis]. AB - Severe acute pancreatitis is morphologically characterized by an extensive and prolonged pancreatic and retroperitoneal inflammation with surimposed patchy or generalized areas of necrosis and hemorraghe in the pancreas and surrounding tissues. Clinical hallmarks include the early development of remote organ dysfunctions, notably cardiorespiratory failure and the late emergence of local complications (hemorraghe, acute pseudocyst and, most importantly, infection). The severity of the attack and the outcome of the patient are critically dependent on the presence and the extent of regional necrosis and are closely related to the casual bacterial contamination of these devitalized areas. The early identification of severe episodes is of therapeutic interest. This prognostic staging is best achieved with a combination of a set of clinical and laboratory data, initial CT findings and single biochemical indicators. Sequential assessment of severity, using biochemical markers and morphological data, is mandatory in order to monitor the fate of regional necrosis. Intensive care treatment includes supportive care of distant organ failures, prophylactic antibiotics and nutritional support. PMID- 8731737 TI - [Surgical treatment of acute pancreatitis]. AB - The aim of surgical treatment in acute pancreatitis is two-fold: 1. pancreatic necrosis excision; 2. suppression of possible cause. Pancreatic surgery is only indicated in acute necrotic pancreatitis with local or systemic complications. Necrosis excision with abdominal drainage has to be confined to patients with refractory systemic disorders despite intensive care management and in case of infected necrosis proved by positive culture of collection obtained under ultrasound or CT-scan. Necrosis excision can sometimes require iteratives laparotomies. Nevertheless, a 25%-hospital mortality rate remains observed in such cases. Surgery is often necessary in cases of pancreatic abcesses and pseudocysts because of the high failure rate of percutaneous treatment. Sometimes, surgery is also required in cases of haemorrhage, intestinal stenosis or perforation, due to the progression of peripancreatic necrosis. Treatment of biliary pancreatitis includes suppression of the biliary cause by cholecystectomy and common bile duct clearance in some cases. The indication of endoscopic sphincterotomy remains to be established. PMID- 8731738 TI - [Natural history of chronic pancreatitis]. AB - The natural history of alcoholic chronic pancreatitis is well-known. Patients who are the most frequently involved by the disease are 40-50 year old men. About 15,000 patients have chronic pancreatitis in France. The first symptoms occur after 10-20 years of alcohol abuse. Pain is the first symptom in 80% of patients and acute pancreatitis in one third of them. During the 5 first years of the course, pseudocyst or common bile duct compression may occur. Between the 5th 10th years of the course, acute bouts are rare but the risk of pseudocyst and extrahepatic cholestasis remains high. Therefore, patients may still undergo surgery. On the other hand, the proportion of pain free patients increases. After the 10th year of the disease, pancreatic calcifications are frequent and exocrine and endocrine pancreatic insufficiency occurs in the majority of patients. PMID- 8731739 TI - [Medical and endoscopic treatment of chronic pancreatitis]. AB - Analgesics and pancreatic extracts are required for the management of pancreatic pain. The use of pancreatic enzymes supplements is necessary for the management of pancreatic insufficiency. Patients with diabetes should be treated with insulin since endogenous insulin secretion is decreased. Drainage procedures of the pancreatic ducts, drainage of pancreatic cysts and biliary prosthesis are different ways of endoscopic management for chronic pancreatitis and its related complications. Endoscopic drainage procedures of the pancreatic duct include pancreatic sphincterotomy, extraction of pancreatic stones by means of balloons or baskets or extra corporeal shock wave lithotripsy, pancreatic duct endoprosthesis. Clinical and anatomical results are good or fair in about two thirds of the cases but there are not yet controlled studies. Endoscopic management of pancreatic cysts use transmural drainage of cysts in contact with the stomach or duodenal wall or transpapillary drainage in cysts communicating with the pancreatic ductal system. The results and complications of the endoscopic treatment of pancreatic cysts are comparable to those of surgical series. Biliary stenting is an alternative to surgical diversion. Endoscopic management of chronic pancreatitis requires the development of new techniques which should be evaluated in further controlled studies. PMID- 8731740 TI - [Surgical treatment of chronic pancreatitis]. AB - The surgical treatment of chronic pancreatitis has benefited from the progress in imaging, with subsequent improvement of preoperative investigations but the main factor has been the development of surgical procedures. The pancreatectomy with the preservation of the pylorus or duodenum completes the armamentarium of by pass and resective techniques. The classical surgical indications are the local evolutive complications: obstruction of pancreatic duct, common bile duct stenosis, duodenal stenosis, splenic complications and localized extra-hepatic portal hypertension. Other indications are intractable chronic pain and pseudotumoral inflammatory lesions of the head of the pancreas that may be indistinguishable from cancer. In the surgical treatment of chronic pancreatitis, the operative mortality is similar with exploratory laparotomy, pancreatic by pass, cysto-anastomosis and the Whipple procedure. 5 and 10 year actuarial survival rates are 92% and 76% with resections, 80% and 54% with cysto anastomosis, 85% and 66% with pancreatic by-pass, respectively. Functional results are excellent with the absence of pain in 76% of patients following resection or by-pass. PMID- 8731741 TI - [Intracranial hypertension. Physiopathology, diagnosis, emergency treatment]. PMID- 8731742 TI - [Mitral valve insufficiency. Etiology, physiopathology, diagnosis, development]. PMID- 8731743 TI - [Neuroleptic drugs. Principles and rules of use]. PMID- 8731744 TI - [Rheumatic pelvispondylitis. Etiology, diagnosis, development, prognosis, treatment]. PMID- 8731745 TI - [Gastroesophageal reflux. Physiopathology, diagnosis, development, treatment]. PMID- 8731746 TI - [Acute diarrhea in infants. Diagnostic orientation (and treatment)]. PMID- 8731747 TI - [Bacterial cutaneous infections: erysipelas. Etiology, diagnosis, development, treatment]. PMID- 8731748 TI - [Correct application of anti-pneumococci vaccination in France]. PMID- 8731749 TI - [Measurement of nasal transepithelial potential difference: a diagnostic test for cystic fibrosis]. AB - Measurement of nasal transepithelial potential difference allows the exploration of transepithelial ionic transports in vivo. Cystic fibrosis is an interesting indication of this test. Indeed, this disease is characterized by a chloride and water secretion deficit across respiratory epithelium. We have measured nasal potential in 8 healthy volunteers. Measurements were repeated 3 times a day, during 3 days for each subject. The reproducibility of the data was analysed with factorial variance model. The mean nasal potential in the healthy volunteers group and in 10 patients with cystic fibrosis was compared. In the cystic fibrosis group, the nasal potential was measured 3 times with a 2 mn-interval between the measurements. No significant variation of the nasal potential values was found from day to day or in the same day from one measurement to another. Mean value was -19 +/- 3.5 mv in normal subjects and -42.6 +/- 5.1 mv in cystic fibrosis patients. We conclude that nasal potential measurement is an easy and reproducible test that might be a complementary tool routinely used along with the classical tests in the diagnosis of cystic fibrosis. PMID- 8731750 TI - [Current aspects of spinal tuberculosis. Apropos of 10 cases]. AB - Between 1990 and 1993 we observed 10 cases of spinal tuberculosis. Magnetic resonance imaging largely contributed to diagnosis and detection of early neurological complications. Medical treatment is essential and must be given for a sufficient duration. Indications for surgery are exceptional and are usually reserved for severe neurological complications. PMID- 8731751 TI - [From first symptom to diagnosis and from diagnosis to treatment of tuberculosis: still a long delay]. AB - OBJECTIVE: The aim of this investigation was to determine the delay between the first symptom and diagnosis of pulmonary tuberculosis and onset of treatment. STUDY DESIGN: Fifty-two consecutive patients presenting with confirmed pulmonary tuberculosis over an 18 month period (June 1, 1991 to December 31, 1992) at the Cochin Hospital in Paris. RESULTS: The mean delay between first symptom and diagnosis was 2 weeks in 14 patients (27%), 3 to 4 weeks in 10 patients (19%) and over 4 weeks in 28 patients (54%). The delay between diagnosis and onset of treatment was less than 8 days in 39 patients. CONCLUSION: Delay was not shorter when symptoms were present. This situation could have an effect on tuberculosis morbidity. PMID- 8731752 TI - [An opacity in the aorto-pulmonary window]. PMID- 8731753 TI - [Pneumococcal pneumonia in a patient with HIV infection. Development to excavation. Apropos of a case]. AB - An HIV-infected patient presented with pneumococcal pneumonia with abscess formation. The causal effect of the pneumococcal infection on abscess formation was discussed. Since the advent of antibiotics, necrotizing pneumococcal pneumonia is rarely observed but clinicians should nevertheless be aware of this possibility, particularly because of the HIV epidemic. PMID- 8731754 TI - [Fibrosing pneumopathy caused by labetalol]. AB - A diffuse interstitial pulmonary fibrosis occurred in a 74-year-old woman treated for ten years with labetalol for systemic hypertension. Bronchoalveolar lavage (BAL) revealed lymphocytic and neutrophilic alveolitis. The lack of another etiologic factor and the spontaneously favorable course after withdrawal of the drug support the hypothesis of a labetalol induced pulmonary fibrosis. PMID- 8731755 TI - [Primary localized bronchopulmonary amyloidosis]. AB - A 56-year-old patient with an uneventful history was hospitalized for investigation of recent dypnea. The physical examination was normal and chest X ray showed localized interstitial lesions in the upper right lobe. Endoscopy showed evidence of diffuse infiltration of the bronchial mucosa and narrowing of the lumen. Transbronchial histology showed evidence of bronchopulmonary amyloidosis. There were no signs of extrapulmonary localizations. Search in other organs was negative demonstrating the primary nature of the lung disease. Primary bronchopulmonary amyloidosis is rarely encountered. PMID- 8731756 TI - [Pleural amyloidosis. Apropos of a case and review of the literature]. AB - The main respiratory manifestations of amylosis are tracheobronchial involvement and modular or diffuse parenchymal disease. Amyloid deposits in the pleura are exceptional. We observed transsudative pleural effusion and heart failure in a patient with multiple myeloma, leading to the discovery of pleural amylosis. Amyloid deposits in the pleura may be fortuitous discoveries since pleural effusion is not necessarily observed. It is however important to be aware of this possibility since the localization is easily accessible for diagnosis. Specific stains are effective diagnostic tools even for transsudative effusions, particularly in suggestive clinical conditions such as multiple myeloma. PMID- 8731758 TI - [Osteoplastic bronchopathy]. PMID- 8731757 TI - [Spontaneous hydro-pneumothorax disclosing malignant mesothelioma. Apropos of 2 cases]. AB - We report the cases of two males who presented with spontaneous complete unilateral pneumothorax with ipisilateral liquid effusion. Neither had a history of previous respiratory disease. In both cases chest tube drainage resulted in recurrence of pneumothorax with chronic illness requiring surgical exploration. The surgery revealed a malignant pleural mesothelioma by histological examination. Thus, spontaneous pneumothorax, particularly with abondant effusion can be a revealing symptom of malignant pleural mesothelioma. PMID- 8731759 TI - Recent developments and rationale towards new strategies for malarial chemotherapy. AB - The major problem facing world research for new antimalarials lies in encountered difficulties in the search for new promising paths. The past 20 years have witnessed a very impressive increase in our understanding of the biochemistry and molecular biology of malaria parasites, with attention focused on specific parasite molecules that are keys to the parasite life cycle or the induction of its pathogenesis. Directed pharmacology research has involved the identification and characterization of targets that can be specifically pharmacologically affected, including the replicating machinery of the parasites, various metabolisms such as the purine salvage pathway, and biosynthesis of pyrimidines or phospholipids. Protease inhibitors (e.g. those degrading haemoglobin), the use of iron chelators or inhibition of heme polymerization, induction of oxidative stress or inhibition of antioxidant enzymes are also investigated. Some pathways have already been validated with current antimalarials but, due to the development of resistance, complete characterization of the molecular structure of the target should allow attack of these exceptional molecules at novel and distinct sites with new drug concepts. The problem in the quest to develop new antimalarials is the fact that the results are not being materialized, but there is no lack of pharmacological targets. PMID- 8731760 TI - The fate of the filaria Litomosoides sigmodontis in susceptible and naturally resistant mice. AB - The fate of Litomosoides sigmodontis was compared in susceptible BALB/c and resistant B10D2 mice, presenting the same major histocompatibility complex (H 2d), with an attempt to dissociate the different elements of the life cycle in order, later, to dissociate the different mechanisms involved. Each female mouse was inoculated once with a small dose of infective larvae (25 L3) or a large dose (100 or 200 L3). In total, 92 BALB/c and 49 B10D2 were studied. Necropsies were performed up to D85 following infection with 25 larvae. The early fate was similar in B10D2 and BALB/c mice; particularly the recovery rate of worms was almost identical during the first month p.i. and represented a quarter of the inoculated larvae. Resistance in B10D2 mice appeared progressively, as judged by retardation of growth and of the fourth moulting, the presence of very small sterile female worms and male worms with abnormal left spicule, and a high frequency of live filariae coated with inflammatory cells and encapsulated dead worms. The L. sigmodontis life span in B10D2 was about half that in BALB/c. Necropsies were carried out up to D20 following infection with 100-200 L3. The recovery rate was increased in BALB/c. Growth was retarded earlier in B10D2 mice, this crowding effect already apparent at D10; this may indicate a role for metabolic factors. The pattern of the life cycle in both mouse strains confirms recent conclusions on Onchocercinae: the recovery rate is established as soon as the second day during "phase 1 of massive destruction", then it is stable during "phase 2 of insignificant mortality". During phase 1, the infective larvae are immediately destroyed in the subcutaneous tissue if they are not able to escape the inflammatory process by penetrating in local lymphatic vessels. By contrast, phase 2, which is longer than the duration of the third larval stage, indicates there is no mortality linked to the third moulting, at least following a single inoculation. PMID- 8731761 TI - [Combinations of ivermectin and diethylcarbamazine for improved control of lymphatic filariasis]. AB - Since 1993, a three arm double-blind controlled trial was implemented in French Polynesia to compare the efficacy of single doses of diethylcarbamazine (DEC) 6 mg/kg vs. ivermectin (IVR) 400 micrograms/kg vs. the combination IVR 400 + DEC 6 for treatment of Wuchereria bancrofti mf carriers. Of the 57 treated male patients, three groups of 19 were randomly selected and allocated to one of the three treatments. Twelve months after the first then the second treatment (M12 then M24) respectively, 3 (16%) then 10 (53%), 3 (16%) then 5 (26%), 7 (27%) then 10 (53%) of patients were mf negative in groups DEC 6, IVR 400 and IVR+DEC. Mf percent return was significantly lower in the group IVR+DEC (2% at M12 then M24) than for the groups DEC 6 (15% at M12 then 4% at M24) and IVR 400 (12% at M12 then 10% at M24). The combination IVR+DEC showed promising results for an annual strategy; to achieve the best results from a control program it is necessary to associate IVR+DEC. PMID- 8731762 TI - Specific and rapid detection of Microsporidia in stool specimens from AIDS patients by PCR. AB - Two microsporidian species, Enterocytozoon bieneusi and Encephalitozoon intestinalis, are the cause of diarrhoea and wasting syndrome in AIDS patients. A new PCR assay is proposed for the rapid and specific detection of these parasites in stools. PMID- 8731763 TI - Therapeutic trial of ivermectin against Notoedres cati var. cuniculi infection in rabbits. AB - Effect of 400 micrograms/kg of ivermectin against natural infection of Notoedres cati var. cuniculi mange in rabbits was evaluated in twenty rabbits which showed typical symptoms of notoedric mange and were randomly divided into two groups. Group I consisted of 15 rabbits treated with single s/c injection of ivermectin (Ivomec, Dynamic Pharmacals, Bombay). In group II, five infected untreated rabbits were kept as control. Daily observations of clinical improvement and on the basis of examination of skin scrapings on days 0, 3, 6, 9, 16, 23 and 30 post treatment were done. Complete visual shedding of lesions was seen on day 6 after treatment and skin scrapping were found negative for mites after 7th day of treatment till the end of experiment. Histopathologically mites were present in stratum corneum and many were noticed in the burrows in epidermis. Marked hyperkeratosis, hyperplasia and acantholysis along with ballooning degeneration of epithelial cells of epidermis were seen. PMID- 8731764 TI - [Cataract surgery after bone marrow transplantation. Apropos of 36 cases]. AB - OBJECT: Bone marrow transplantation has improved the prognosis of haematological malignancies, but patients are consequently at considerable risk of developing ocular complications, such as cataracts. MATERIAL AND METHODS: A retrospective case study of 36 eyes (24 patients), with cataract extraction after the transplantation. Surgery was performed at a mean of 4.5 years after transplantation and the average age was 33 years at the time of surgery. The lens changes were all of the posterior subcapsular type. Surgery was indicated for symptoms of dazzling and glare; visual acuity was only moderately decreased (4/10, P6). RESULTS: Cataract surgery required specific measures such to "polish" the posterior lens capsule, since cataract changes were often very adherent. The surgery follow-up was uncomplicated with the exception of five eyes which had an inflammatory reaction (3 cyclitic membrans, 2 "pseudo-endophthalmitis") but which showed complete regression after treatement. The follow-up after surgery varied from 6 months to 4.5 years. Final visual outcome was improved. 82.5% achieved 8/10, and 96.8% achieved P2. Three YAG-laser capsulotomies were performed, mainly for posterior capsulofibrosis. COMMENTS: Cataract surgery after bone marrow transplantation appears to have a good prognosis. Posterior capsule "polishing", though difficult, is possible in most cases. In our study, secondary cataract appears to be less frequent, but, further long-term follow-up is needed to confirm this statement. PMID- 8731765 TI - [Results of cataract surgery in patients with uveitis]. AB - PURPOSE: Extraction of cataracts with posterior chamber intraocular lens (PC-IOL) implantation has been attempted with some encouraging results in uveitis having minor intensity. However, the number of operated patients is very limited. We present the results in patients having more important intensity of uveitis. MATERIALS AND METHODS: The authors studied 28 eyes in 20 patients who underwent cataract extraction as a complication of uveitis. RESULTS: All the operations were accomplished by phacoemulsification and aspiration (PER) with posterior chamber lens implanted in 23 eyes. Etiologic diagnosis included 8 sarcoidosis, 5 Behcet, 3 Vogt-Koyanagi-Harada syndrome and 12 other aetiologies. Eighty percent of patients obtained visual acuity better than 0.6 postoperatively. Postoperative complications were not severe. CONCLUSIONS: The indication of PC-IOL could be given to cataracts in chronic uveitis if the patients are older than 35 years and there are no uveal inflammatory signs at least 3 months prior to considered implantation. PMID- 8731766 TI - [Risk factors of postoperative proliferative vitreoretinopathy in giant tears]. AB - BACKGROUND: In eyes with giant retinal tears, the rate of severe postoperative PVR and failure to permanently reattach the retina remains especially high in spite of technical advances in surgical management. This study was conducted to elucidate the clinical and surgical risk factors for severe postoperative PVR in such eyes. PATIENTS AND METHODS: We reviewed the records of 68 consecutive patients (69 eyes) with giant retinal tears. Univariate and multivariate statistical analyses were used to evaluate the risk factors for severe PVR. RESULTS: The rate of severe postoperative and failure to permanently reattach the retina were 43.5% (30/69 eyes). It was influenced at a statistically significant level by two independent risk factors: 1) the presence and severity of preoperative PVR and 2) the use of cryotreatment as compared to the use of ALP treatment. Severe postoperative PVR occurred in 63.6% (14/22 eyes) of eyes managed with cryotreatment versus 31.1% (14/45 eyes) of eyes managed with ALP treatment (P < 0.02). The rate of severe postoperative PVR was 64% (16/25 eyes) in eyes with grade C-D PVR preoperatively versus 31.8% (14/44 eyes) in eyes with no PVR or grade B PVR preoperatively (P < 0.01). In eyes managed with the use of ALP treatment the rate of severe postoperative PVR remained influenced at a statistically significant level (P < 0.005) by the presence of grade C-D PVR preoperatively. Grade C-D PVR was significantly more frequent preoperatively in patients with visual symptoms of 3 week-duration or more at initial examination (23/24 patients, 95.8%), than in those with visul symptoms under 3 week-duration (8/41 patients, 19.5%) (P: 0.0005). CONCLUSION: The results suggest that the high incidence of severe postoperative PVR in giant retinal tears may be decreased by 1) early management before the occurrence of PVR and 2) the use of argon laser photocoagulation rather than cryotreatment as the method of creating a chorioretinal scar. PMID- 8731767 TI - [Topical anesthesia for cataract surgery in adults]. AB - PURPOSE: This prospective study compares the analgesia obtained by topical anaesthesia versus a single peribulbar anaesthesia. The comparison is made during clearcornear phacoemulsification in adult patient. METHODS: Ninety-six eyes are operated with Lidocaine 2% (66) or Tetracaine 1% (30) topical anaesthesia, 38 eyes received an unique anterior periocular injection. The same surgeon performs the phacoemulsification in 2 different centers. Analgesia is evaluated in the per and post operative period by the surgeon and a nurse. RESULTS: Four cases (4.2%) in the topical anaesthesia group required a complementary subconjonctival anaesthetic injection. Seven cases (7.3%) had pain during surgery but without any need for a complementary injection. In the other group 1 injection was necessary (2.5%). A slight pain was felt by 5% of patients during surgery. CONCLUSION: Topical anaesthesia is reliable but needs a patients selection, a well trained staff and a non beginner surgeon. The advantages are obvious for out patient surgery if no premedication is associated. Many doctors will continue to ask for an akinesia during cataract surgery even is the analgesia is obtained. PMID- 8731768 TI - [Responsibility of incomplete retinal photocoagulation in the indication of vitrectomy in proliferative diabetic retinopathy]. AB - PURPOSE: We report an epidemiologic study about prophylactic photocoagulation of ischaemic diabetic retinopathy in order to avoid proliferative stage and severe vitreous haemorraghe. METHODS: We studied retrospectively 42 eyes of 32 diabetic patients who underwent a vitrectomy in 1993. For each patient several items were analyzed: nationality, sex, age, type, duration and clinical course of diabetes, and other risk factors. Slit lamp and ocular fundus examination were noted; in particular lens, iris and vitreoretinal framework. RESULTS: Almost all patients had an inadequate laser photocoagulation: in 26 eyes (62%) it was insufficient or non-existent, uncomplete in 12 eyes (29%), and non confluent for 18 eyes (43%). Only four eyes have had complete and confluent panphotocoagulation. CONCLUSION: Photocoagulation appears to be the priority therapy to avoid most severe complications of proliferative diabetic retinopathy. Laser photocoagulation must be started from preproliferative retinopathy, confluent on the useless ischaemic retinal areas, and dense elsewhere. PMID- 8731769 TI - [Effects of human recombinant basic Fibroblast Growth factor on endothelial wound healing in organ culture of human cornea]. AB - PURPOSE: The effects of human recombinant bFGF has been evaluated on 9 paired human donor corneas (age 75 +/- 8 years), preserved in organ culture medium. MATERIAL AND METHODS: The endothelium of corneas were mechanically wounded (area of 7.53 +/- 2.05 mm2) and placed in culture medium during 14 days. For each pair, one cornea was tested with bFGF (50 ng/ml), delivered in two times (day 0, day 7), according to the stockage of the bFGF on the basal membranes (low affinity receptor), while the ipsilateral cornea served as control. Endothelium was assessed by trypan staining at day 0, day 7, and day 14. At this term of fourteenth day, alizarine red and trypan blue staining permitted morphometric data. RESULTS: The bFGF factor increases significantly cell density in the wound area (p < 0.05), and in the transitional area (p < 0.01), compared to the control group. In the transitional area, cells depletion was only 15% (392 +/- 55 cells/mm2) in the treated group compared to the 28% (716 +/- 0.1 cells/mm2) in the untreated group. In the wound area, the mean cell area was averaged 2581 microns2 in the control group and 2161 microns2 in the bFGF treated group (p < 0.05); in the transitional area the mean cell size was 549 microns2, and 479 microns2 in the control and the bFGF treated group (p < 0.05) respectively. The bFGF group do not affect the shape factor. CONCLUSION: This assay demonstrates that human bFGF greatly facilitates wound closure in endothelium of human cornea. The cellular migration from the transitional zone seems the dominant healing mechanism. PMID- 8731770 TI - [Lamellar autografting in the treatment of small corneal perforations]. AB - BACKGROUND: The treatment of corneal perforations due to corneal ulceration is a serious challenge in countries where there are no eye banks. The author presents an original surgical procedure for treating these cases: lamellar autografting. METHODS: The corneal transplant was taken at 12 h; size was 2 to 4 mm and thickness 2/3 of the stroma. Twelve patients were treated. RESULTS: The results are satisfactory and the grafting successful; no complication of significant importance has been noted. CONCLUSION: Lamellar autografting is an effective method for treating small corneal perforations. It can be temporary in case of central localization, or permanent in case of paracentral perforation. PMID- 8731771 TI - [Unilateral congenital cataract and visual function]. AB - The authors report the case of a young patient presented with cytomegalovirus infection involving the nervous, respiratory and ocular systems. A congenital monocular cataract associated with a chorioretinal macular scar in the controlateral eye imposed surgical treatment of the cataract. No retinal damages were found in the cataractous eye after surgery. Three years later the visual function is supported by the phakic eye in spite of the relative precocity of surgery (4th month), and of the correct antiambliopic treatment. This case would suggest that the period in which a monocular cataract can be treated with good functional results, should be restricted within the first few weeks of life. PMID- 8731773 TI - [Dry syndrome and cataract surgery. A case]. PMID- 8731772 TI - [Central serous chorioretinopathy in a pregnant woman]. AB - PURPOSE: A case of central serous chorioretinopathy in a 28-year-old pregnant woman is reported. METHODS: A complete ocular examination was performed including fundus biomicrospic examination and fluorescein angiography. RESULTS: Examination of the left eye revealed central serous chorioretinopathy associated with greyish white subretinal exudate. In the right eye, sequaele of asymptomatic central serous chorioretinopathy was found. Reattachment of the retina and dissolution of the exudate, with return of visual acuity to normal, occurred soon after delivery. CONCLUSION: This case report is the 19th case of central serous chorioretinopathy during pregnancy reported to date. Central serous chorioretinopathy in pregnant woman is often associated with subretinal exudation which is probably fibrinous in nature. The affection resolves spontaneously at the end of pregnancy or after delivery, but may recur in the context or outside of subsequent pregnancy. The special conditions of pregnancy, including haemodynamic, biological and psychological alterations may lead susceptible women to develop central serous chorioretinopathy. PMID- 8731774 TI - [Endophthalmitis. 1: diagnosis]. PMID- 8731775 TI - [Epidemiological study of 1705 malignant tumors of the eye and adnexa]. AB - PURPOSE: Malignant tumours of the eye and adnexa are generally rare, and in the absence of a French national register of declaration of cancers, only an approximate evaluation can be made. We studied the epidemiology of tumours of the eye and adnexa from the laboratory results of anatomical pathology of biopsies examinated at the laboratory of Centre Hospitalier National d'Ophtalmologie des Quinze-Vingts in Paris. MATERIALS AND METHODS: Statistical analysis of the results of histological diagnoses, their sites and the personal histories recovered from the laboratory registers from 1925-1995, a period of 70 years, were carried out. RESULTS: 1705 malignant tumours of the eye and adnexa diagnosed over 70 years consisted of 760 melanomas (44.6%), 450 carcinomas (26.4%), 338 retinoblastomas (19.8%), 95 haematopoetic tumours (5.6%), and 62 diverse others (3.6%). These tumours were localised in: the endo-ocular (1035 cases), lids (398 cases), conjunctiva (119 cases), orbit (90 cases), lacrimal glands (23 cases), caruncle (10 cases), eye lashes (8 cases), optic nerve (6 cases), cornea (3 cases), lacrimal sac (2 cases) and undefined (11 cases). CONCLUSION: In our study, endo-ocular tumours are the most frequently seen of all tumours of the eye and adnexa (60.7%) and are essentially represented by melanomas in the elderly and by retinoblastomas in the newborn and infants. Carcinomas are the most frequent lid malignant pathology whose presence increases with age. Tumours originating from haematopoetic tissues are often seen in the sub-epithelial tissues of orbit, lids, conjunctiva and lacrimal gland. PMID- 8731776 TI - [Changes in adrenergic innervation of the choroid during aging]. AB - BACKGROUND: Aging causes loss of neurons and adrenergic nerve termination in various human organism systems and apparatus. The aim of this work is the observation of qualitative and quantitative differences in the orientation and density of the human choroid adrenergic innervation in relation to age. METHODS: The choroid specimens were taken from the corresponding sectors of the eyeballs of four patients aged between 70 and 75 years and patients aged between 40 and 45 years. The glyoxilic method of Furness and Costa (1975) was used. RESULTS: The location and orientation of the adrenergic fibres was similar in the adult and aged patients. However, the fibres are clearly fewer in number, less rich in varicosity and have reduced fluorescence. In the suprachoroid lamina thickness and in the vascular layer, the adrenergic synaptic contacts with the melanocytes are fewer in number, as are also the SIF-cells (small intensely fluorescent cells). CONCLUSIONS: With age there is a significant reduction of the orthosympathetic innervation of the human choroid. This fact could explain some physiopathological aspects of the choroid circulation and blood flow and, therefore, play a part in ocular pression variations. PMID- 8731777 TI - [Treatment of vitreoretinal proliferation in rhegmatogenous detachment and silicone oil tamponade]. AB - PURPOSE: To describe our technique of vitrectomy and silicone oil tamponade for managing retinal detachment and to report the last results according to the posterior and anterior proliferative vitreoretinopathy. METHODS: A retrospective study was conducted in 108 patients who underwent vitreoretinal surgery and silicone oil tamponade for proliferative vitreo-retinopathy, 64% patients had already been operated without success and 42% underwent vitrectomy with SF6 or C3F8. Diffuse posterior proliferative vitreo-retinopathy (grade C3-D) was present in 64% patients and anterior proliferative vitreoretinopathy was present in 43.5%. Silicone oil was removed in 79% patients after a mean duration of 6.3 months. It was replaced by 16% C3F8. All patients were followed for a minimum of 6 months. RESULTS: Before silicone oil removal, 55% of the retinas were reattached posterior to the scleral buckle with one operation, 78% after 2 operations and 88% after 3 or 4 operations. An average of 2.1 vitrectomy surgeries were performed. Reproliferation was correlated with the anterior proliferative vitreo-retinopathy (p < 0.001), posterior proliferative vitreo retinopathy (p < 0.01) or previous vitrectomy (p < 0.05). The final visual acuity was 1/20 or better in 61% of the eyes and 2/10 or better in 30.5%. After silicone oil removal, 8% retinas redetached, 21% of the eyes had hypertony, 7.5% had hypotony and 7% of the corneas had dystrophy. CONCLUSION: Silicone oil tamponade was effective for the treatment of retinal detachments with proliferative vitreo retinopathy. Since the main complications were hypertony and corneal dystrophy, silicone oil should be reserved for severe proliferative vitreo-retinopathy cases. PMID- 8731778 TI - [Prediction of functional recovery after surgery of macular epiretinal membranes. Value of interferometry using Lotmar's apparatus]. AB - BACKGROUND: The predictability of the measurement of the macular function by white light interferometry was evaluated in a group of patients whose epiretinal membranes had been surgically removed. METHOD: Fifteen patients presenting either primitive or secondary epiretinal membranes were included. We used the apparatus described by W. Lotmar. The main evaluation criteria was the best corrected visual acuity (VA) chosen between the third month and the sixth month examinations. RESULTS: Preoperatively the mean visual acuity was 0.227 +/- 0.105. The mean final visual acuity was 0.507 +/- 0.198. The final visual acuity was predicted within an 0.1 interval in ten cases (67%). In two cases the predicted visual acuity was identical to the pre-operative VA and the final results confirmed the test. No improvement occurred. CONCLUSION: The Lotmar visometer is statistically predictive of the visual outcome after epi-retinal membranes surgery. PMID- 8731779 TI - [Magnetic resonance imaging in the diagnosis of doubtful cases of small melanoma of the choroid]. AB - The main complementary procedures used to diagnose a choroidal melanoma are A- and B-mode ultrasonography, fluorescein angiography and magnetic resonance imaging (MRI). With MRI the malignant tumour has unique MRI signature owing to the paramagnetic properties of melanin. MRI can thus help in making the differential diagnosis and in estimating tumour extension. The advantage of MRI is that it provides complementary information that, when added to the results of the other investigative techniques, results in a quasi certain diagnosis. Three clinical cases are discribed. PMID- 8731780 TI - [Radial keratotomy for the treatment of Excimer laser undercorrections]. AB - BACKGROUND: Effectiveness of radial keratotomy for undercorrections following photorefractive keratectomy. METHODS: Twenty myopic eyes from minus 4.75 to minus 8.25 were treated by Excimer laser photorefractive keratectomy and regressed to a mean myopic refraction of minus 2.20 +/- 0.81 after a follow-up of 10 to 22 months (mean: 16.25). Radial keratotomy was performed using Hoffmann Fyodorov Thornton nomogram. Blade depth was set according to pachymetry performed before surgery under microscopic examination, paracentrally on the meridian of each incision. RESULTS: Postoperative results were evaluated after 6 to 20 months follow-up. After radial keratotomy mean refractive error was minus 0.30 +/- 0.43 and minus 0.32 +/- 0.25 at 3-6 and 6-12 months respectively. Visual acuity, without correction, was prior to radial keratotomy: 2.18 +/- 0.99; 8.125 +/- 0.6 at 3-6 months and 8.43 +/- 1.57 at 6-12 months after radial keratotomy. CONCLUSION: Radial keratotomy appears to be a safe, simple and predictible technique to treat undercorrections following Excimer laser photokeratectomy. PMID- 8731781 TI - [Value of cyanoacrylate tissue adhesives in peripheral corneal ulcers of inflammatory origin]. AB - PURPOSE: To investigate cyanoacrylate tissue adhesives in ulcerative keratitis. They represent one of the best emergency treatments of corneal descemetoceles, preperforated corneal ulcers, impending corneal perforations and small corneal perforations. METHODS: Nine patients presenting with peripheral ulcerative keratitis were treated by this method between 1990 and 1994. The underlying diseases included Mooren's ulcer (5 eyes), rosacea (2 eyes) and rheumatoid arthritis (2 eyes). RESULTS: Despite the poor prognosis of ulcerative keratitis, good healing and corneal re-epithelialization were obtained in 6 cases (66%) without any relapse. Severe infectious complications occurred in 1 case (11%) and required a therapeutic penetrating keratoplasty. CONCLUSIONS: Cyanoacrylate tissue adhesives are an effective and well tolerated treatment of peripheral ulcerative keratitis associated with ocular inflammatory diseases. Potential infectious complications impose a strict follow-up. PMID- 8731782 TI - [Biodegradable controlled-release 5-FU implant in the surgery for glaucoma. Experimental study]. AB - BACKGROUND: 5-Fluorouracil (5-FU) is useful as an adjunct treatment for glaucoma filtering surgery. However, efficacy depends upon maintaining sustained drug levels, currently achieved by repeated daily injections of the drug for several days. To overcome this limitation, we designed a biodegradable implant for the sustained release of 5-FU. MATERIAL AND METHODS: The implant (0.79 mm diameter, 6 mm long, 2.2 mg weight) contains 0.66 mg of 5-FU and is loaded in a needle coupled to a custom-made instrument to permit subconjonctival insertion of the implant through a 2 mm wide snip incision. The in vitro and in vivo pharmacokinetics as well as the biocompatibility studies of the implants were assessed in the rabbit. RESULTS: No infection, inflammatory reaction or extrusion occurred. The implant released the drug at a constant rate of 1.20 micrograms/hr for over 18 days and totally biodegraded in less than 86 days. CONCLUSION: Implantation of the device, after laser sclerostomy or conventional trabeculectomy shows great potential for the treatment of glaucoma. To determine the implant's efficacy, additional studies in the cat are underway. PMID- 8731783 TI - [A case of conjunctival raspberry tumor of viral origin]. AB - We describe the clinical and histopathological findings of a conjunctival papilloma due to papillomavirus. The therapeutic method applied was excision plus cryotherapy. These lesions are known to recur, and papillomavirus DNA sequences are present in several cases of conjunctival dysplasia and carcinoma. Using biological molecular methods (in situ hybridization, Southern blot hybridization, and particularly polymerase chain reaction), more than 60 Human Papillomavirus have currently been reported. This new detection techniques improve the possibility of findings HPV infection, and increase the interest for this virus extremely widely distributed in man. PMID- 8731784 TI - [A familial form of epidermoid carcinoma of the conjunctiva]. AB - Two two-sided cases of conjunctival epidermoid carcinoma and one dysplasia in the same family are reported. The three sisters concerned were teenagers. This familial disease with bilateral localizations in young female patients, is quite rare. Clinical, histological and epidemiological conclusions were based on familial investigation. PMID- 8731785 TI - [Edematous decompensation of the cornea after silicon implant of the posterior chamber in phakic eyes in myopia]. AB - The authors report a case of a girl operated with a silicon IOL in the posterior chamber for the correction of myopia in phakic eye. One year after surgery there was a total bilateral corneal decompensation which required corneal transplantation. The possible hypothesis of the damage are discussed. PMID- 8731786 TI - [Isolated ocular recurrence of relapsing polychondritis. Apropos of a case]. AB - Ocular manifestations of relapsing polychondritis occur in 60% of patients, most often in association with other systemic manifestations of the disease. Episcleritis is the most common manifestation, but scleral perforation, retinal vasculitis, optic neuritis and necrotizing scleritis can lead to blindness and require the use of immunosuppressive agents. We report the case of a 72-year-old woman with diffuse bilateral scleritis as the single manifestation of relapsing polychondritis. High dose steroids were used with success. PMID- 8731787 TI - [Para-papillary chorioretinitis. A case]. PMID- 8731788 TI - [Multidrug resistance and its reversal. General review of fundamental aspects]. AB - Among the mechanisms by which cancer cells evade chemotherapy, multidrug resistance (MDR) is certainly the best known. MDR is characterised by cross resistance between numerous natural products used in cancer treatment, especially antibiotics and plant alkaloids. MDR results from a defect in cell accumulation of the drugs, which are actively effluxed from cells by a plasma membrane pump, which is a high molecular weight glycoprotein termed P-glycoprotein. This protein is encoded by a gene called mdr1, and can be inhibited by a variety of pharmacological compounds. The activation of the mdr1 gene can occur via numerous types of stimulation, especially anticancer drugs themselves, which can induce mdr1 gene transcription. P-glycoprotein is an ATPase transporter which is believed to extrude xenobiotics from the plasma membrane rather than from cytoplasm. Although potential sites of interaction of P-glycoprotein with its various ligands have been identified, especially at the level of putative transmembrane domains, the exact mechanism for drug pumping has never been elucidated. Reversal of MDR in vitro is easy to obtain and to characterise. An important development aims at identifying substances able to reverse MDR in the clinical setting, that are devoid of any pharmacological properties other than interaction with P-glycoprotein. Other targets can be postulated for these MDR modulators, whose combination could well lead to a synergistic reversal of drug resistance. PMID- 8731789 TI - [Evaluation of multidrug resistance phenotype on medullary specimens from patients with acute leukemia by determination of nuclear efflux of tetrahydropyranyl-doxorubicin. Approach by confocal laser microspectrofluorometry]. AB - Confocal microspectrofluorometry allows the analysis of fluorescent molecules such as anthracylines in isolated living cells. An optical microscope fitted with a phase-contrast 100 X water-immersion objective enables simultaneous observation of the sample, focusing of the laser beam on the selected cell fraction (nucleus) and collection of the fluorescence emitted from the sample. The resulting intranuclear spectra are interpreted according to a quantitative model of the fluorescence spectra of both free and DNA-bound anthracycline. The intranuclear drug concentration can thus be determined. This technique has been applied to blast cells collected in patients with acute leukemia. Leukemic cells are aspirated from bone marrow, separated by Ficoll sedimentation and resuspended in RPMI-1640 containing 10% fetal calf serum and 200 nM tetrahydropyranyl doxorubicin (THP-DOX). After one hour, 20 cells are analyzed and the mean nuclear drug content is determined (C1). Cells are then resuspended in the same medium but without anthracycline for 3 hours and the mean intranuclear drug concentration is then also determined (C3). From C1 and C3 the retention rate (RR) is calculated. Firstly, the accuracy of the method was checked. In 4 AML patients, two different samples aspirated on the same day were divided into two portions. Thus, two measurements were made on each one (4 values per patient). Coefficients of variation were satisfactory (4, 6, 12, and 12%). Secondly, blast cells collected in patients with AML and ALL at diagnosis or in relapse were studied. P-glycoprotein (P-gp) and CD34 expression was also studied using respectively immunohistochemistry land flow cytometry. Results obtained from the first 21 patients showed that there was no correlation between RR and either P-gp or CD34 expression. This could result from the efflux of THP-DOX by other mechanisms and/or low sensitivity of the staining technique. PMID- 8731790 TI - [Predictive value of intracellular accumulation of daunorubicin and P glycoprotein expression simultaneously determined by flow cytometry in adult acute myeloid leukemias]. AB - Multidrug resistance (MDR) phenotype expression was evaluated retrospectively in 87 patients with acute myeloid leukemia (AML), 69 with de novo AML, ten with relapsed AML and eight with AML secondary to myelodysplastic syndrome (MDS). MDR phenotype, characterized by P-glycoprotein expression (MRK16 monoclonal antibody) and decrease in intracellular daunorubicin (DNR) accumulation was determined using flow cytometry. All patients received chemotherapy including cytosine arabinoside and anthracycline (daunorubicin, zorubicin, idarubicin) or mitoxantrone, and quinine in ten cases. The predictive value of the MDR phenotype for clinical responsiveness was studied using uni- and multivariate analyses. Univariate analysis showed that DNR accumulation (p < 10(-4)), P-glycoprotein expression (p = 10(-4)) and disease status (de novo versus recurrent AML and acute MDS) (p = 10(-4)) were predictive of clinical responsiveness. The significance of these three parameters was maintained in multivariate analysis. When de novo AML was considered, only DNR accumulation was of predictive value (p < 10(-4)) for complete response to chemotherapy. PMID- 8731791 TI - [Effect of S9788, cyclosporin A and verapamil on intracellular accumulation of doxorubicin, daunorubicin and daunorubicinol in primary rat hepatocyte culture]. AB - The aim of this study was to compare the effect of three MDR modulators, cyclosporine A, S9788 and verapamil on the efflux of two anthracyclines, doxorubicin and daunorubicin and of daunorubicinol, the C-13 alcohol metabolite of daunorubicin. Rat hepatocyte primary cultures were used as a model of P-gp expression. They allow to study MDR at different levels of P-gp expression which increases in parallel with culture time. Furthermore, hepatocytes are able to metabolize drugs and enable determination of the role of P-gp on metabolite efflux. Hepatocytes grown for 4 or 48 hours were incubated for 6 hours in the presence of a combination of each modulator and one of the two anthracyclines (0.5 microM). Modulator concentrations used were 1, 5 and 15 microM when associated with DOX, and 1 and 15 when associated with DNR. In fresh hepatocytes, the three MDR modulators did not induce an increase in intracellular retention of the two anthracyclines compared to controls without MDR modulators. At 48 hours of culture, the three tested drugs increased intracellular accumulation of DOX. However, daunorubicin retention was not modified but that of its metabolite was increased. The activity rank order was cyclosporine A > S9788 > verapamil. Cyclosporine A and S9788 were active in simultaneous as well as in sequential combinations with anthracyclines. Verapamil was only effective when co-incubated with anthracyclines. PMID- 8731792 TI - [Modulation of chemoresistance: methodology of therapeutic trials]. AB - A certain percentage of cancers are primarily or subsequently resistant to chemetherapeutic agents. Several biological mechanisms are implicated in this phenomenon, including multidrug resistance/P-glycoprotein (mdr1/P-gp), resistance related proteins (P-95 and P-110), multidrug resistance associated protein (P 190), iso-enzymes of gluthatione S-transferase, topo-isomerases, glutathione peroxidase and others. mdr1/P-gp overexpression has been studied in many types of cancer. It represents an inducible, transferable and phylogenetically ancestral biological system. It is expressed at the surface of the cell, and in that way, it participates to several normal functions. The recent introduction of modulators/revertants of mdr1/P-gp may change some concepts in using chemotherapy for cancers. The first step is represented by a better knowledge of the cancers which overexpressed mdr1/P-gp, with determination of the best biological technique, including the gold standards. This allows the clinician to clarify the best impact of such a therapeutic way and to define the criteria of modulator selection. Such criteria includes in vitro selection using a panel of sensitive/resistant cell lines, in vivo tests including transgenic mice, nude or SCID mice, and toxicological studies. Choice of modulated drug is easier and depends on the biological target. For mdr1/P-gp, major drugs included doxorubicin and vinca-alkaloids. Due to the fact that some modulators have an influence on the pharmacokinetic parameters of chemotherapeutic drugs, it is important to verify such parameters. The last choice concerns the strategy of drug development with three levels of action: 1) modulation of clinical chemoresistance, intrinsic or acquired one; 2) modulation of biological resistance; 3) leading to the prevention of the amplification of low levels of chemoresistance. A new therapeutic way is born, which takes care of a dynamic aspect of the tumor, and necessitates a new use of chemotherapy. PMID- 8731793 TI - Detection of the multidrug resistance of P-glycoprotein in healthy tissues: the example of the blood-brain barrier. AB - The blood-brain barrier is formed by the cerebral capillary endothelial cells, joined together by tight junctions. These cells express the general endothelial cell markers as well as specific markers found on endothelial cells forming physiological barriers such as gamma-glutamyltranspeptidase, the glucose transporter Glut1 and the neutral amino-acid transporter. Using the monoclonal antibodies C219 and MRK16, we have revealed by Western blot and immuno histochemistry the expression of the multidrug resistance P-glycoprotein on isolated rat cerebral cortex capillaries. On the other hand, P-glycoprotein was not detectable in brain cortex homogenates. P-glycoprotein thus appears to be a blood-brain barrier endothelium-specific marker which could regulate brain penetration of xenobiotics and thus participate in the neuroprotection of the brain. PMID- 8731794 TI - [Risk factors of the occurrence of cerebral toxoplasmosis in patients with HIV infection. Definition of indications of primary prevention. Proposals of the Toxoplasmosis and other protozoan infections Group. Coordinated action N.5 - 1996 February]. PMID- 8731795 TI - [Future of bone allografts in massive bone resection for tumor]. AB - Currently cryopreservation offers the best means of preserving bone tissue for allografts. At -196 degrees C all enzyme activity is halted and tissue preservation is unlimited. Perfect sterilization, adapted cryoprotection and controlled freezing and thawing are now part of the well-controlled process of bone preservation in bone banks. At implantation, the mechanical properties of cryopreserved bone is as good as or better than fresh bone, although the diaphyseal cortical bone is more fragile. Anatomic and physiologic reconstruction of the graft area is a major factor in graft resistance. It takes approximately three weeks for vessels to penetrate cancellous allografts and at least one month for total revascularization. The delay may reach several years for cortical bone. The immune response of the host is determined by the antigenic properties of the leukocytes in the bone marrow as well as the blood vessels and nerves. The protein-mineral complex itself has little antigenic effect. A certain number of non-specific immune reactions result from transfusions almost always performed with bone grafting. At the present time, there is no artificial material capable of providing a mechanically acceptable substitute for allograft bone in limb reconstruction. Allograft bone currently stocked in bone banks provides a biologically and clinically acceptable means of reconstruction after major bone loss. Other factors such as public acceptance and administrative authorizations will also play an important role in the future of massive reconstruction with bone allografts. PMID- 8731796 TI - [Value of an extensive biological study in venous or arterial thromboses]. AB - OBJECTIVES: The aim of this work is to study the signification of an extensive biological evaluation in patients with "unexplained" thrombosis. We studied 78 patients with more than one arterial and/or venous thromboembolic event. METHODS: Fifty-four patients were admitted for unexplained deep venous thrombosis (group I, n = 19, 9 men and 10 women) and/or arterial thrombosis (group II, n = 35, 21 men and 14 women). A third group (group III) included 24 patients (13 men, 11 women) known to have a pathologic state which can lead to a thrombotic event. RESULTS: The patients in both groups I and II had, more often than normal subjects, a high level of homocysteinemia (26% vs 3%, p < 0.001), anti-beta 2 glycoprotein 1 (18.5% vs 3%, p < 0.001) and antiphospholipid antibodies (13% vs 3%, p < 0.02). We also found a significant association between an increase of erythrocytic aggregation and arterial thrombosis (group II). In the third group, for both arterial (n = 14) and venous (n = 10) thrombosis, we found a high level of anticardiolipin antibodies (25% vs 3%, p < 0.001), anti-beta 2 glycoprotein 1 antibodies (12.5% vs 3%, p < 0.05) and abnormal erythrocytic aggregation (16.5% vs 3%, p < 0.01). In these 3 groups the other studied parameters (Lp(a), platelet aggregation, cryoglobulin, cryofibrinogen, antinuclear antibodies, anticytoplasm antibodies, plasma and urine immunoelectrophoresis, protein C, protein S, antithrombin III, plasminogen) were not different from levels observed in normal subjects. CONCLUSION: An extensive biological analysis, including plasma homocystein level, anticardiolipin antibodies, anti-beta 2 glycoprotein 1 antibodies and a study of the erythrocytic aggregation would appear to be of value in patients presenting recurrent arterial or venous thromboembolic events. Specific therapy can be applied in case of abnormal results continued anticoagulant therapy for anticardiolipin and anti-beta 2 glycoprotein 1 antibodies, and a vitamin therapy for increased homocysteinemia. PMID- 8731797 TI - [Auto-dialysis: an 11-year experience of a hemodialysis center in France]. AB - OBJECTIVES: Report 11 years of experience with self-managed hemodialysis in patients medically apt for extra-hospital dialysis and living close enough to small outpatient hemodialysis units to become totally self-sufficient. RESULTS: Among the 276 patients with chronic renal failure managed at the hemodialysis center at the Troyes hospital during the 11-year study period from 1984 through 1994, self-managed hemodialysis at small outpatient units was initiated in 127 (46%). None of these 127 patients required medical assistance or specific care during dialysis sessions. At initial hospital admission, only 60/127 (47%) were totally self-sufficient: 52 (41%) were later graft recipients; and 21 (16.5%) had to return to the hospital for a medical or surgical condition incompatible with extra-hospital care but all of these 21 patients remained self-sufficient. CONCLUSION: Extra-hospital hemodialysis in units close to the patients residence offers patients a better quality of life, even when medical assistance is required. All patients who require hemodialysis can thus be treated at lower cost without compromising quality of treatment. Perfect self-sufficiency may not be a goal in itself, but self-managed hemodialysis can be a very useful technique for patients without major medical problems. Continuing contact with the organizing hemodialysis center guarantees the safety of the system. PMID- 8731798 TI - [Retrospective evaluation of the detection of Toxoplasma gondii by polymerase chain reaction in AIDS patients]. AB - OBJECTIVES: We retrospectively evaluated routine detection of Toxoplasma gondii by polymerase chain reaction (PCR) amplification of the B1 gene and the TGR 1E sequence in blood and CSF samples from patients with acquired immune deficiency syndrome (AIDS) and suspected toxoplasmosis. METHODS: From January 1993 to February 1994, 93 blood, 33 cerebrospinal fluid (CSF) and 1 aqueous humor samples were obtained from 83 HIV-positive patients with CD4 counts under 200/mm3 and suspected toxoplasmosis. RESULTS: Authentic cerebral toxoplasmosis was confirmed by response to specific treatment in 18/29 patients with typical focal brain lesions. Blood samples were available in 17/18 and CSF in 6. PCR was positive for both B1 and TGR (23.5% sensitivity, 100% specificity) in 4/17 blood samples and for either B1 or TGF (58.9% sensitivity, 72.7% specificity) in 10/18. PCR of CSF was positive in only 1/6 patients with cerebral toxoplasmosis. PCR (TGR 1E only) was positive in 3/11 blood samples and in one CSF sample from patients without cerebral toxoplasmosis. Five out of 21 patients with diffuse neurologic symptoms and presumed HIV encephalitis had positive Toxoplasma detection in blood or CSF. However, no clinical improvement was obtained after specific antitoxoplasmic therapy. Two out of 38 patients with unexplained fever with or without pneumonia had a proven disseminated toxoplasmosis. These two patients had PCR-positive blood samples. One of them was cured by a specific anti-toxoplasmic treatment and the other died two days later. Seven other patients had a positive PCR result in blood or CSF. Three of them improved with specific treatment and 2 died before treatment could be initiated. The one patient with toxoplasmic retinitis was PCR positive both in blood and aqueous humor. CONCLUSION: PCR detection of Toxoplasma gondii appears to add little to the diagnosis of cerebral toxoplasmosis but could be helpful in the diagnosis of cerebral toxoplasmosis associated with extracerebral reactivation and in disseminated toxoplasmosis. PMID- 8731799 TI - [Ventricular extrasystole and arrhythmogenic right ventricular dysplasia. Critical analysis of the diagnostic value of non-invasive tests]. AB - Asymptomatic ventricular extrasystoles were discovered in 2 active sportsmen (32 and 33 years). The cardiology work-up rapidly led to the diagnosis of arrhythmogenic right ventricular dysplasia in one. In the other, the clinical presentation was similar and the initial diagnosis was ventricular extrasystole with a healthy heart. Four years later however, the subject was still asymptomatic but a second evaluation revealed arrhythmogenic right ventricular dysplasia. This delayed diagnosis emphasizes the importance of renewed noninvasive evaluation of patients with asymptomatic ventricular extrasystoles. PMID- 8731800 TI - [Hemodynamic management of septic shock. From a global approach to a gastrointestinal approach]. AB - Oxygen uptake and increased metabolic requirements are characteristic of the acute state resulting from septic shock. One therapeutic approach to improving hemodynamics is to increase oxygen delivery in an attempt to overcome tissue oxygen debt. Recent randomized trials have suggested that systematically increasing oxygen supply is not necessarily the ideal strategy. It might be better, for each patient, to reach an optimal oxygen supply. This requires identification of parameters capable of indicating the optimal level, a rather difficult task. Currently, focus has been placed on the importance of the splanchnic circulation in severe septic shock. Hypoxia resulting from hypoperfusion of the intestinal mucosa occurs early in sepsis and could, via intermediary bacterial and/or endotoxinic translocation, maintain the septic syndrome and favor development of multiple organ failure. Since the drugs used to restore hemodynamics have vasoactive properties, measuring their effect on relevant indicators of splanchnic perfusion and oxygenation such as PCO2 or pH within the gastric mucosa using tonometric gastric probes might be a means of determining optimal oxygen level. With this approach, it would be possible to avoid sacrifying the perfusion of the gastrointestinal mucosa by using drugs which appear to favor the microcirculation in this territory. PMID- 8731801 TI - [Myopericarditis and atrial fibrillation disclosing leptospirosis septicemia]. PMID- 8731802 TI - [Neuroendocrine carcinoma of the lymph nodes without primary tumor. 2 cases]. PMID- 8731803 TI - [Salmonella typhimurium cerebral abscess: rare complication of HIV infection]. PMID- 8731804 TI - [Tumor graft on the tract of the catheter of percutaneous gastrostomy inserted by endoscopic approach in a patient treated for inoperable cancer of the esophagus]. PMID- 8731805 TI - [Isolated paralysis of the common oculomotor nerve disclosing multiple sclerosis]. PMID- 8731806 TI - [Cytomegalovirus pneumopathy and hemolytic anemia in an immunocompetent patient]. PMID- 8731807 TI - Germany: the medical profession and the structural development of the health care system. PMID- 8731808 TI - Austria: some aspects of the health workforce. PMID- 8731809 TI - Belgium: an overview of physician manpower policy issues. PMID- 8731811 TI - Spain: an explanatory model of supply and utilization of human resources of the health care system. PMID- 8731810 TI - Denmark: demography of doctors. PMID- 8731812 TI - Finland: health workforce in the early 90's. PMID- 8731813 TI - [France: the spirit of laws or the uncertain number of physicians]. PMID- 8731814 TI - Greece: the state of health workforce. PMID- 8731815 TI - Ireland: medical manpower in public hospitals. PMID- 8731816 TI - Italy: towards privatization of employment and European qualification of health manpower. PMID- 8731817 TI - Luxembourg: medical manpower development. PMID- 8731818 TI - Netherlands: cost containment and worforce salaries. PMID- 8731819 TI - Portugal: current distribution and development of the human health resources. PMID- 8731820 TI - United Kingdom: managing medical supply or balancing different agendas. PMID- 8731821 TI - Sweden: recent developments in physician manpower. PMID- 8731822 TI - [Apoptosis and blood disorders]. PMID- 8731823 TI - [A clinical study on multidrug resistance gene (MDR1) expression in acute leukemia]. AB - The authors established a highly sensitive, specific and quantitative method-RT PCR for measuring the levels of MDR1 mRNA in 91 acute leukemic samples (including 30 untreated cases, 32 remission cases, 29 refractory and relapse cases). The results showed that MDR1 mRNA positive rate for refractory and relapse cases, untreated cases and remission cases were 82.8%, 40.0%, 28.1% respectively. In untreated patients, it was found that the first complete remission rate differed significantly between MDR1 positive (41.7%) and MDR1 negative groups (88.9%) (P < 0.05). In remission group, MDR1 positive cases had a higher relapse rate than MDR1 negative case (66.7%; 13.0% P < 0.01) and all these cases relapsed of MDR1 gene overexpression (6 cases) were resistant to further treatment. It is concluded that the expression of MDR1 mRNA might be an unfavorable prognostic factor for patients with acute leukaemia and could predict the efficacy of intensive chemotherapy and serve as a high risk factor for early and resistant relapse. PMID- 8731824 TI - [A study on the peripheral blood stem cells mobilization, collection and their effects on engraftment]. AB - The methods of mobilization and collection of stem cells in peripheral blood stem cells transplantation (PBSCT) and the association between the number of stem cells transplanted and hematopoietic recovery were studied. The investigation was carried in 22 patients (11 acute leukemia, 6 multiple myeloma, 4 non-Hodgkin's lymphoma, 1 breast cancer). Three regimens for mobilization were carried out as follows: 1) chemotherapy + tetrahydrofolic acid + dexamethasone, 2) chemotherapy + rhGM-CSF + dexamethasone, 3) chemotherapy + rhG-CSF + dexamethasone. Besides, CD34/CD33 dual-color direct immunofluorescence flow cytometry assay was performed in 7 cases in the rhG-CSF group. The results showed: 1) The mean number of collected cells (MNC) in the rhG-CSF group was MNC (8.29 +/- 6.14) x 10(8)/kg and CFU-GM (21.35 +/- 17.24) x 10(4)/kg, being highest among the 3 groups. 2) The number of CD34+ cells correlated with MNC and CFU-GM. CD34+ cells in the peripheral blood were 0 or < 0.5% before mobilization and increased markedly 6-8 days after rhG-CSF administration. Harvesting should be started at that time and carried out every day until CD34+ cells reached 5 x 10(6)/kg. 3) The number of PBSC transplanted was the key to hematopoietic recovery. PMID- 8731825 TI - [The clonal origin of multiple myeloma]. AB - In order to explore the clonal origin and malignant pathway of multiple myeloma (MM), a study has been made to investigate the differentiation stage of malignant progenitor cells in peripheral blood (PB) of MM. The immnophenotype of mononuclear cells (MC) in PB and bone marrow (BM) from 17 patients with MM were studied with APAAP method by using monoclonal antibodies directed to a series of B-cell markers (CD10, CD19, CD20, CD45RA, CD45RO, CD38). There were no plasma cells in samples of PB examined. Out of the 17 patients, MC of PB from 10 expressed plasma cell antigen CD38. This expression was significantly different from normal control (P < 0.001) and identical with that of BM. MC of 9 patients were CD38+ and CD45RO+, 3 were CD38+, CD45RO+ and CD45RA+ and 1 was CD38+, CD45RA+, CD45RO+ and CD10+ in PB. The results indicate that while the B-cell marker's expression of PB matches with that of BM, its forms are diversified. This suggests that the PB of MM patients contains precursors at multiple differentiation stages of myeloma cells, from early B cell to active B cell, late B cell and preplasma cell. MC of PB from 10 patients expressed CD45RO while that of BM from 5 out of them expressed the same. This indicates that preplasma cells which are CD45RO+ in PB may eventually return to BM for homing and differentiating into plasma cells, thus losing CD45RO and expressing CD38 only. This study has proved the presence of malignant cells in PB of MM, confirmed the speculation of the malignant pattern and provided the theoretical basis for purging of PB in autologous peripheral blood stem cell transplantation. PMID- 8731826 TI - [Tuberculosis in chronic renal failure patients with or without renal replacement therapy]. AB - Tuberculosis developed in 40 out of 978 chronic renal failure (CRF) patients with or without renal replacement therapy during a five-year period. The incidence was 4.1% and was 10 times higher than that in the general population. The onset of symptoms was within 1 year after dialysis or kidney transplantation in 68% of the patients. 82.5% of them had predominantly extrapulmonary tuberculosis and most of these were lymph node infection. The diagnosis of tuberculosis in these patients was difficult; tuberculin sensitivity was present only in 6.4% of the patients and examination of sputum and other fluid for mycobacteria was usually unrewarding. Detecting serum anti-PPD-IgG level might be a valuable supplementary diagnostic method, because 85% of patients with active tuberculosis showed positive results. Measuring serous exudate and urine anti-PPD-IgG level by ELISA or M. Tuberculosis DNA by PCR might be helpful in the diagnosis of tuberculous serositis and urinary tract mycobacterial infection. PMID- 8731827 TI - [A study on protein metabolism in nephrotic patients treated with Chinese herbs]. AB - It was found in our previous studies that two Chinese herbs Astragali and Angelica (A&A) together with high protein diet could ameliorate the lowering of serum albumin level and increase the synthesis rate of protein as shown by 15N glicine tracer priming protein turnover study in nephrotic rats. Further experiment was designed to investigate the role of A&A and high protein intake in protein dynamic study and nitrogen balance in nephrotic patients. The level of serum total protein (STP), serum albumin (SA), urinary protein loss (UP), serum cholesterol (Cho) and index number of protein turnover and nitrogen balance in 7 patients were measured before and after treatment of 30 days with A&A. The results showed that after treatment the patients had significantly increased STA and SA (44.3 +/- 5.60 vs 49.7 +/- 6.80 P < 0.01; 22.6 +/- 0.42 vs. 29.4 +/- 7.40 P < 0.05), decreased UP and Cho (6.54 +/- 1.83 vs 4.63 +/- 1.33 P < 0.05; 9.69 +/ 2.31 vs. 7.82 +/- 1.95 P < 0.05) and increased net rates of total protein synthesis (1.06 +/- 0.03 vs 1.27 +/- 0.12 P < 0.05). It is concluded that A&A together with high protein intake could improve the disorder of protein metabolism and increase the level of serum protein by improving the net rate of protein synthesis in nephrotic patients. PMID- 8731828 TI - [Point mutation of p53 and detection of human papillomavirus DNA in bronchogenic carcinoma]. AB - The p53 gene has been implicated in the pathogenesis of lung cancer as a tumor suppressor gene. Aberrations of the p53 gene (exon 6-8) was examined in 34 surgical specimens of lung cancer with single-strand-conformation-polymorphism analysis of polymerase chain reaction products. Structural abnormalities of the p53 gene were observed in 30 tumor specimens (88.2%), i.e, in 10 of the 12 specimens with squamous cell carcinoma, 8 of the 10 with adenocarcinoma, all the 3 small cell carcinoma, the only one large cell carcinoma, all the 5 alveolar cell carcinoma and all the 3 adenosquamous carcinoma. The aberrations of the p53 gene were not limited to a particular histological type or clinical stage and were not associated with degree of differentiation or history of heavy smoking. It is suggested that high mutation rate of the p53 gene may participate in the genesis of lung cancer. In this study, multiple primer polymerase chain reaction method was applied to detect the human papillomavirus DNA (HPV DNA) in human lung cancer. Only 4 of these 34 specimens were HPV DNA positive, and they were all of squamous cell carcinoma, 3 of them had p53 mutation. PMID- 8731829 TI - [In vitro antibacterial activity of cefoperazone-sulbactam]. AB - This study was conducted to evaluate the antibacterial activity of CPZ-SBT in 1,146 clinical isolates and compared with that of CPZ and other antimicrobial agents. CPZ-SBT has much better antibacterial activity than CPZ against B lactamase producing organisms. Of the 834 gram negative organisms tested, 165 were CPZ resistant, but 94 (57.0%) of them were still susceptible to CPZ-SBT, CPZ SBT broadens the antibacterial spectrum of CPZ, it has good activity against Acinetobacter spp and B fragilis. Compared with other antimicrobial agents tested, CPZ-SBT is as active as ceftazidime, amikacin and ciprofloxacin against Enterobacteriaceae, P aeruginosa and Acinetobacter spp, but slightly less active than imipenem. It is as active as imipenem, timentin and metronidazole against B fragilis and other anaerobes. The results show that CPZ-SBT is a new member of the broad spectrum antimicrobial agents. It may become a promising agent for the treatment of severe infections caused by cefoperazone-resistant Gram negative bacilli including P aeruginosa. PMID- 8731830 TI - [A randomized controlled clinical trial of sulperazone as compared with cefotaxime in the treatment of bacterial infections]. AB - A randomized controlled clinical trial of sulperazone as compared with cefotaxime was conducted. 207 patients with bacterial infections entered the study. The overall clinical efficacy rate of sulperazone and cefotaxime was 95.15% and 90.38% respectively. The bacterial clearance rate in the two groups was 84.7% and 80.6% respectively. The adverse drug reactions in both groups were mild with the rate of 7.77% and 8.65% respectively; there was no statistical difference between the two groups. The results of disc susceptibility test showed that the sensitive rate of the clinical isolates to sulperazone was 90.9%, being significantly higher than that of cefotaxime 69.3% (P < 0.001). PMID- 8731831 TI - [Clinical effects of bisoprolol in congestive heart failure due to dilated cardiomyopathy]. AB - Bisoprolol was administered orally at doses of 1.25-5 (mean 3.2 +/- 1.1)mg/day for 3-10 (mean 4.1 +/- 1.6) months in 19 patients with congestive heart failure secondary to dilated cardiomyopathy. All patients heart rate and blood pressure were sensitive to the first dose 1.25 mg bisoprolol and when the dose was titrated to 2.5 mg, it was effective and led to a smooth course during a 10-hour period of observation. After treatment, the patients heart function, including fractional shortening, ejection fraction and exercise ability increased significantly and both the severity and the incidence of ventricular arrhythmias decreased significantly. The response of heart rate to exercise and isoproterenol infusion tended to be normal lymphocyte beta-adrenergic receptor density was increased. It is shown that bisoprolol is a promising beta-blocking agent in treating congestive heart failure. The mechanisms of its effects may be related to depression of excessive sympathetic activity, decrease of heart rate and up regulation of beta-adrenergic receptor. PMID- 8731832 TI - [An approach to the diagnosis of recurrent attack of rheumatic fever in patients with rheumatic heart disease]. AB - In order to evaluate recurrent attack of rheumatic fever (RARF) in patients with rheumatic heart disease (RHD), 97 patients with diagnosis of RHD for two years and more were included in this study. Progression of the lesions of cardiac valves was found in every case by comparing the echocardiographic and Doppler findings at present and two years ago. The history of the initial attack of acute rheumatic fever or the appearance of RHD were reviewed. The present complaints at admission and necessary laboratory procedures were studied. The data showed that 75 of the 97 patients had history of initial attack of ARF and the remaining 22 were symptomless with indolent carditis or carditis of insidious onset two years ago. In this admission, only 25 of the 97 patients had ARF in the Jones criteria were followed strictly, while the remaining 72 did not fulfill the criteria. It is suggested that if there is evidence of recent group a streptococcal infection, the conditions for diagnosis of RARF are as follows: 1) When one major or more than two minor criteria are present in a patient with RHD, a definite diagnosis of RARF may be made. 2) A presumptive diagnosis of RARF may be made, when a patient presents one minor criteria and several other manifestations such as anemia, abdominal pain, rapid sleeping pulse rate, tachycardia out of proportion to fever, malaise, epistaxis, precordial pain and an elevated level of IgG, IgA, C3 and circulating immune complexes. PMID- 8731833 TI - [Evaluation of the combined assay of alpha-feto protein and gamma-glutamyl transpeptidase isoenzyme II in the diagnosis of primary hepatic carcinoma]. AB - Serum alpha-feto protein (AFP) and gamma-glutamyl transpeptidase isoenzyme II (gamma-GTP-II) were assayed in 104 patients with primary hepatic carcinoma (PHC), gamma-GTP-II had a higher specificity of 95.7%, while AFP a higher sensitivity of 84.6%. There was no significant difference in the positive diagnostic rate of the two markers in patients with AFP > or = 400 micrograms/L and with AFP < 400 micrograms/L. Nor was there any difference if the patients were divided into four groups according to the serum AFP levels of 200 micrograms/L, 400 micrograms/L and 1,000 micrograms/L. Altogether 97 of the 104 PHC patients were detected when the two markers were assayed in combination (40 out of 42 PHC patients with AFP > or = 400 micrograms/L and 57 out of 62 PHC patients with AFP < 400 micrograms/L). These results suggest that combined assay of the two serum PHC markers will increase the detecting rate of PHC and be of value especially in those patients with normal or low serum level of AFP. PMID- 8731834 TI - [Detection of T-antigen in colorectal adenocarcinoma and polyps]. AB - Galactose oxidase method was employed to detect the beta-D-Gal (1-->3) -D-Gal NAc residue of T-antigen present in the large intestinal mucus of 156 subjects. The positive rates of the test were 84.4%, 29.1%, and 7.2% in the mucus samples obtained from 32 patients with colorectal adenocarcinomas, 55 with polyps and 69 controls respectively. Chi-square test demonstrated that there were significant differences between the group of carcinoma and control (P < 0.001) as well as between also polyp and control (P < 0.01). The test had a high sensitivity (84.4%) and specificity (92.8%) in the diagnosis of colorectal cancer and may be used as a practical mass screening test for colorectal neoplasms. PMID- 8731835 TI - [Anti-hepatitis C virus screening to prevent hepatitis C virus infection in blood donors]. AB - Before carrying out anti-hepatitis C virus (anti-HCV) screening in blood donors, 433 plasma donors and 461 whole blood donors from 4 county plasmapheresis units and 5 city blood centres in Hebei province were surveyed. Anti-HCV positive rates were 55.5% and 10.2% in the two populations respectively. The anti-HCV positive rates also differed between qualified and unqualified donors. In qualified and unqualified plasma donors, they were 46.2% and 91.6% respectively and in qualified and unqualified whole blood donors 5.8% and 42.3% respectively. One year after anti-HCV screening, 793 plasma donors and 873 whole blood donors in 3 city blood centres and 4 county hospitals were surveyed, the anti-HCV positive rates in plasma and whole blood donors were 4.0% and 0.2% respectively. Compared with those before screening, there were 91.3% and 96.6% reduction respectively. The anti-HCV positive rate in voluntary donors was 0.5%. All the 86 first time donors were negative for anti-HCV before and after screening. PMID- 8731836 TI - [Apoptosis and leukemia]. PMID- 8731837 TI - [Hormones and apoptosis]. PMID- 8731839 TI - [The new outlook of histiocytes and histiocytosis]. PMID- 8731838 TI - [Disseminated intravascular coagulation and multiple system and organ failure]. PMID- 8731840 TI - [Immunohistochemical study of hepatitis C virus (HCV) antigen in liver tissue of patients with HCV infection]. AB - Hepatitis C virus antigen (HCV Ag) and HCV-NS3 protein in paraffin-embedded liver tissue of patients with HCV infection were detected with immunohistochemical method by using polyclonal anti-HCV and monoclonal anti-HCV-NS3 for studying the expression of HCV in liver tissue. The results showed that the detection rate of HCV Ag and HCV-NS3 were 38.3% and 30.0% respectively. All the specimens with HCV NS3 positive tissue were also those with HCV Ag positive. The stained substance located in the cytoplasm of liver cell, and the stained cells distributed scattered, clustered or diffuse in liver lobular. No obvious infiltration or necrosis was observed around the positive cells. There was obvious difference of the detection rate between different models of serum HCV markers. These results provided some help in understanding the pathogenesis of HCV. PMID- 8731841 TI - [Expression of proliferating cell nuclear antigen in the liver tissue of hepatitis B patients]. AB - Liver biopsy and autopsy specimens of 153 cases, including 39 cases of acute severe hepatitis (ASH), 21 cases of subacute severe hepatitis (SSH), 11 cases of chronic severe hepatitis (CSH), 22 cases of active cirrhosis (AC), 20 cases of acute mild (AMH), chronic persistent (CPH) and active hepatitis (CAH), respectively, were immunohistochemically stained for proliferating cell nuclear antigen (PCNA) to compare the proliferative activity of hepatocytes by using labelling index (LI). LI is higher in CAH, SSH, and AMH than in AC and CSH (P < 0.01). In the latter, LI was completely negative in most of the specimens (25/33, 75.8%) and few scattered hepatocytes positive for PCNA were found in the remaining 8 cases (8/33, 24.2%) in the area next to the fibrotic septum and varied greatly in different pseudolobules. The results showed that the proliferative activity was very low in the AC and CSH. In some cases of ASH, the residual hepatocytes still kept prominent proliferating ability; it meant that the survived percentage would have been increased with regeneration of hepatocytes if those patients had not died early. In ASH and SSH, LI was significantly higher in patients who survived than in those who died (P < 0.01). The detection of PCNA in liver tissue with severe viral hepatitis is useful for prognostic evaluation. PMID- 8731842 TI - [Genetic variation in the cleavage site of the precore region of hepatitis B virus in Chinese patients with fulminant hepatitis]. AB - In this study, a previously not documented variant of hepatitis B virus was described in Chinese patients with fulminant hepatitis. The entire precore/core region amplified from samples was cloned into a bacterial vector and sequenced by dideoxy chain termination reaction. Double amino acid substitutions were seen in the precore region in the isolates: one from glycine to aspartic acid at codon 29 previously reported; the other substitution of phenylalanine for valine at codon 17 in the cleavage site of hepatitis B virus. Loss of hepatitis B virus e antigen in these patients with fulminant hepatitis might therefore be due to the mutation in the cleavage site, rather than the emergence of a stop codon in the precore region of hepatitis B virus. Accumulation of hepatitis B e antigen precursor within the hepatocytes might account for the fulminant hepatitis exacerbation. PMID- 8731843 TI - [Multiple analysis for the c-Ha-ras activation and DNA content in human gastric precancerous lesions]. AB - The relationship between ras activation and over expression of p21 protein and DNA content in progression from benign to malignant of gastric mucosa was studied. The point mutation at 12 of c-Ha-ras oncogene was detected with PCR and specific oligonucleotide probe hybridization, the expression of ras p21 and DNA ploidy by flow cytometry was also carried out simultaneously on 66 human gastric tissues with gastric cancer and precancerous lesions, some of the patients were followed up for 42 to 60 months. The results showed that: activation of ras oncogene and p21 overexpression may be involved in the early stages of gastric cancer; DNA aneuploidy may not help for the early diagnosis, but can be useful for the diagnosis of gastric cancer in pathology; gastric cancer with mutational activation at 12 of c-Ha-ras may has unfavourable future. It concluded that c-Ha ras activation and p21 overexpression may be help to early diagnosis of gastric cancer in clinic. PMID- 8731844 TI - [The role of endoscopic ultrasonography and epidermal growth factor receptor assay in preoperative staging for gastric carcinoma]. AB - Endoscopic ultrasonography (EUS) was performed in 36 patients with gastric carcinoma and the epidermal growth factor receptor (EGFR) of the specimen from endoscopic biopsy in 19 patients with gastric carcinoma was measured with immunoassay. Results of EUS were compared to the pathology of resected specimens according to the new (1987) TNM classification. EUS was very accurate in assessing the depth of tumor infiltration and the lymph node metastasis. Overall accuracy in T and N stage were 81% and 72% respectively. The level of EGFR in patients with infiltration into the neighbouring organs (97.7 +/- 27.9 fmol/mg pro.) were significantly higher than those without infiltration (47.7 +/- 21.3 fmol/mg pro.) (P < 0.01). The EGFR levels of those with lymph node metastasis (85 +/- 24.3 fmol/mg pro.) were significantly higher than that without lymph node metastasis (37.1 +/- 16.1 fmol/mg pro.) (P < 0.01). The results show that EUS as well as the EGFR assay could be used to determine the progress and prognosis of gastric carcinoma. PMID- 8731845 TI - [Association of plasma insulin with blood pressure and sodium sensitivity in children]. AB - To study the relationship between plasma insulin level, sodium sensitivity and blood pressure, 150 children aged from 10 to 15 years with high blood pressure and 150 children matched for age and sex but with normal blood pressure were measured for plasma insulin with radioimmunoassay and sodium sensitivity with the tests of oral saline load and furosemide sodium-volume depletion and followed for 5 years. The results showed that (1) In children with HBP and positive family history of hypertension (FH+), the plasma insulin level was significantly higher than that in the respective control group (P < 0.01); (2) In children with high plasma insulin level, the percentiles of systolic blood pressure kept high and had a rising tendency during the follow-up period; (3) In children with sodium sensitivity, the plasma insulin level was higher than that in children with sodium resistance (P < 0.01) and the plasma insulin level correlated positively with blood pressure increasing rate of post-load (P < 0.05). It is suggested that insulin resistance was associated with sodium sensitivity, and they might play a role in the development of hypertension in childhood. PMID- 8731846 TI - [Left ventricular diastolic dysfunction in patients with angina decubitus]. AB - Our study population consisted of three groups: 1. 24 cases with ejection fraction (EF) > 45% in 26 patients with angina decubitus (AD) were studied as group A. 2. 20 patients had coronary artery disease without AD as group B. Group B and A were matched for age, EF and coronary artery obstructive lesions. 3. 20 cases without cardiovascular diseases as group C. Left ventriculography (LVG) was performed in these patients. The results showed that left ventricular (LV) early 1/3 filling fraction (1/3FF) was significantly reduced in group A as compared with that in group B and C (both P < 0.001). The findings suggest that patients with AD have abnormalities of LV diastolic filling. Left ventricular end diastolic pressure (LVEDP) significantly increased in patients with AD after LVG as compared with before LVG (P < 0.01). However, there was no statistically difference in LVEDP before and after LVG in both group B and C, indicating that the compliance of LV was also decreased in patients with AD. The increase of LV volume after recumbency would cause increase of LV wall tension and myocardial contraction and aggravate the LV diastolic dysfunction, resulting in a progressive increase in myocardial oxygen consumption. Therefore, we consider that on the basis of severe coronary artery obstructive lesions, LV diastolic dysfunction is a significant pathologic factor in the pathogenesis of AD. PMID- 8731847 TI - [Effects of a platelet-activating factor antagonist ONO-6240 on antigen induced eosinophil infiltration into the airways]. AB - An in vivo mouse model of allergic bronchial asthma was developed to investigate the effects of a platelet activating factor antagonist ONO-6240 on eosinophil (EOS) recruitment into the mouse airway. In the present study, no EOS could be found in bronchoalveolar lavage fluid (BALF) from normal control mice. However, chronic ovalbumin challenge of sensitized mice induced significant bronchoalveolar eosinophilia (9.68 +/- 0.72 x 10(8)/L). In three groups of mice treated intraperitoneally with different doses of ONO-6240 (0.1, 1.0, 10.0 mg/kg), the medication produced 42.1% (5.60 +/- 0.97 x 10(8)/L, P < 0.01), 58.8% (3.99 +/- 0.84 x 10(8)/L, P < 0.01), and 72.6% (2.65 +/- 0.71 x 10(8)/L, P < 0.01) inhibition, respectively, of the antigen-induced influx of EOS into BALF. It was found that the prevention of EOS infiltration into airway with ONO-6240 was accompanied by decrements of both interleukin (IL)-5 and IL-2 levels. These data indicated that ONO-6240 prevented airway eosinophilia by inhibiting production of IL-5 and IL-2. Our results suggested that ONO-6240 may be of value in treating human asthmatic patients. PMID- 8731848 TI - [Clinical and angiographic characteristics of renovascular hypertension]. AB - Clinical and angiographic characteristics of renovascular hypertension were studied in 95 patients. The patients were divided into 3 groups: 55 cases with aortic arteritis (group AA), 27 with fibromuscular dysplasia (group FMD) and 9 with atherosclerosis (group AS). The patients in group AS were significantly older in age and had longer history of hypertension. Abdominal bruit was heard significantly more often in group AA and serum potassium was significantly lower in group FMD. Angiographic data showed that in group AA lesions in thoracic aorta and abdominal aorta was found in 81.4% and those involving both renal arteries in 52.5%. In group FMD, 82.5% of patients had lesion in renal artery on one side and none had lesion in thoracic and abdominal aorta. In group AS, lesions were found mainly in thoracic and abdominal aorta, accounting for 77.7% and lesions in renal arteries were mainly unilateral. In group AA, lesions were found in 90 renal arteries altogether. Among them, 58.9% was in the proximal part of the renal artery; the lesion was either localized stenosis (67.8%) or obstruction (17.7%). In group FMD, lesions were found in 33 renal arteries altogether. Among them, 48.5% was in the middle or distal part of the renal artery and 27.3% resembled string of beads. In group AS, a total of 10 renal arteries were involved with 4 (40%) of ostial stenosis and 4 (40%) total obstruction. PMID- 8731849 TI - [A study on the value of vitamin C in treating skeletal fluorosis]. AB - Three hundred and sixteen patients diagnosed as skeletal fluorosis roentgenologically and having no other diseases were divided randomly into 2 groups, matchable in sex distribution, age, functional state and X-ray findings. In the treated group every patient took 200mg vitamin C twice a day, while in the control group placebo tablets. The duration of treatment was 3 months. The therapeutic effect was evaluated by measuring the joint angle and power of gripping and studying the X-ray film and electromyogram. 38 kinds of biochemical parameters were measured simultaneously with a MCA-3 autobiochemical analyser. The results were as follows, (1) The total effective rate in the treated group was 12.84%, chi 2 = 0.556, P > 0.05, so there is no obvious effect in treating skeletal fluorosis with vitamin C. (2) The electromyogram in the treated group showed a recovering rate of 20.00%, chi 2 = 5.9, P > 0.01. It is further shown that there is no effect in treating skeletal fluorosis with vitamin C. (3) 38 kinds of biochemical parameters measured simultaneously and the X-ray findings also showed that vitamin C is not effective. Thus the validity of using vitamin C as the drug of first choice to treat skeletal fluorosis is to be queried. PMID- 8731850 TI - [A randomized control study on the treatment of 123 cases of bacterial infections with cefteram and cefaclor]. AB - To evaluate the efficacy and the safety of cefteram in bacterial infections, a randomized control study of cefteram and cefaclor on the treatment of 123 patients with respiratory and urinary tract infections was carried out. The result showed that the effective and bacterial eradication rates were 92.1% and 91.4% for cefteram. 83.3% and 85.2% for cefalor. Adverse reactions were mainly gastrointestinal reactions, occurring in 4.6% of the cefteram group and 9.4% of the cefaclor group. Study of minimum inhibitory concentration displayed high antibacterial activity of cefteram for enterobacteriaceae and other Gram-negative organisms and its activity was higher than that of gentamyicin and ciprofloxacin for E. coli. It is concluded that cefteram was effective and safe in the treatment of respiratory and urinary tract infections. PMID- 8731851 TI - [Resistance to activated protein C]. PMID- 8731852 TI - [Nitric oxide and hyperdynamic circulation in liver cirrhosis]. PMID- 8731853 TI - [Further recognition on the adult respiratory distress syndrome]. PMID- 8731854 TI - [Polymerase chain reaction technique in the diagnosis of the diseases of internal medicine]. PMID- 8731855 TI - [Changes in serum eosinophil cation protein level and blood eosinophil density in patients with asthma]. AB - To study the functional state as well as the relation-ship between eosinophil and T lymphocyte in the asthmatics, the authors measured the eosinophil density of blood with percoll, the level of serum eosinophil cation protein (ECP) with radioimmunoassay and soluble IL-2 receptor (sIL-2R) concentration with ELISA. The results revealed that the proportion of hypodense eosinophil and the levels of serum ECP and sIL-2R were 36.3% +/- 14.80%, 10.74 +/- 7.22 mg/L and 0.33% +/- 10 U/L respectively in asthmatics, being significantly higher than those in normal subjects (14.71% +/- 8.20%, 6.93 +/- 4.91 mg/L, 0.13 +/- 0.03 U/L, respectively, P < 0.05 in all). There is remarkable correlation between any two of the three parameters (r = 0.40, 0.37, 0.39, respectively, P < 0.05). It is suggested that eosinophil activation is associated with T lymphocyte activation, both may be involved in the pathogenesis of asthma. PMID- 8731856 TI - [A preliminary observation on the effects of inhaled nitric oxide on bronchial asthma]. AB - Nitric oxide (NO) is a vasodilator as well as a bronchodilator. In order to estimate the therapeutic effects of NO on bronchial asthma, 10 patients in acute exacerbation stage were selected for this study. 40 ppm NO was inhaled for 20 minutes. The clinical stage and pulmonary function were evaluated instantly. The results showed that subjective symptoms were relieved and wheezing rales decreased to a different extent, FEV1 and PEF improved significantly (P < 0.05). It is suggested that low concentration NO inhalation can be used as a therapeutic measure for bronchial asthma. PMID- 8731857 TI - [Effects of nasal continuous positive airway pressure on breathing pattern and respiratory drive in patients with obstructive sleep apnea syndrome]. AB - To examine the influence of continuous positive airway pressure (CPAP) therapy on respiratory center drive in patients with obstructive sleep apnea syndrome (OSAS), 20 normocapnic OSAS patients (group 0) and 20 simple snoring patients were studied. In the first night, diagnostic polysomnography (PSG) was performed. Before and after PSG monitoring, mouth occlusion pressure (P0.1), tidal volume (VT), minute ventilation (VE), respiratory rate (RR), inspiratory time (Ti), expiratory time (Te), total cycle duration (Ttot), inspiratory duty cycle (Ti/Ttot), mean inspiratory flow (VT/Ti) and effective inspiratory impedance (P0.1/VT/Ti, Ieff) were measured while they were breathing room air. In the following night the OSAS patients were treated with nasal CPAP and PSG monitoring and the above mentioned measurements were repeated. The results showed that pre PSG values of P0.1, RR and P0.1/VT/Ti in the OSAS patients were significantly higher than those in the snoring patients, while VT, Ti, Te and Ttot values were lower. In the first night, the post-PSG P0.1 value in the OSAS patients increased markedly as compared with the pre-PSG. After overnight nasal CPAP therapy, the respiratory disorder index in the OSAS patients decreased markedly, the nadir SaO2 increased markedly, but the post-PSG P0.1 value did not increase significantly. It is concluded that, before sleep, OSAS patients exhibit a higher respiratory drive and a shallow and frequent breathing pattern as compared with simple snoring patients. After nocturnal sleep, the respiratory drive of OSAS patients increases significantly, the breathing pattern becomes more shallow and frequent. Nasal CPAP may effectively relieve the sleep apnea and hypopnea as well as the resulting hypoxemia and therefore correct the changes in breathing pattern and respiratory drive through nocturnal sleep in patients with OSAS. PMID- 8731858 TI - [The significance of changes in plasma renin angiotensin aldosterone system and atrial natriuretic factor in patients with high altitude myocardial hypoxidosis]. AB - Plasma renin activity (PRA), angiotensin II (ATII), aldosterone (ALD) and atrial natriuretic factor (ANF) were determined in 30 patients with high altitude myocardial hypoxidosis (HAMH) with radioimmunoassay and the data were compared with those of 40 healthy subjects and 26 patients with high altitude chronic cardiopathy (HACC). All of them have been in Lasa, a place with an altitude of 3,658 meter, for a long time. The results showed that PRA and plasma ATII increased significantly (P < 0.05-0.01) and plasma ANF decreased significantly (P < 0.01) in patients with HACC and were higher than those in the control. It is suggested that disturbance of these biologically active substances might play a role in the development of HAMH. PMID- 8731859 TI - [Hemodynamic and renal effects of strong diuretic atrial natriuretic peptide in patients with acute myocardial infarction]. AB - The hemodynamic and renal effects of strong diuretic atrial natriuretic peptide (ANP) were investigated in 12 patients with acute myocardial infarction. Strong diuretic ANP was administered as a bolus injection of 100 micrograms and followed by a continuous infusion of 2 micrograms/kg within 10 minutes into the right atrium via a catheter. We found that strong diuretic ANP induced an increase in cardiac index and stroke volume index. Right atrial pressure, pulmonary arterial pressure, pulmonary capillary wedge pressure, systemic vascular resistance and pulmonary vascular resistance were decreased after ANP infusion. The maximal responses were observed immediately after the infusion was completed. No significant change of arterial blood pressure and heart rate was observed. Furthermore, strong diuretic ANP also increased the urine volume, urinary sodium, urinary chloride and urinary potassium. These results suggest that strong diuretic ANP infusion may induce significant changes of hemodynamics and may be used to prevent the development of severe circulatory volume overload in patients with AMI. PMID- 8731860 TI - [Application of serology and 14C-urea breath test to monitoring of the effect of anti-Helicobacter pylori chemotheraphy]. AB - In order to evaluate the value of serology and 14C-urea breath test in monitoring the effect of anti-Helicobacter pylori chemotherapy, at the first stage of this study, endoscopy, 14C-urea breath test and serology (H. pylori IgG antibody measured by ELISA) were performed in 42 of patients before the onset of anti-H pylori chemotherapy and at 1, 3, 6 and 12 months after termination of the treatment. On analyzing the change of the results of breath test and serology during the one year follow-up period in these patients, "above the cutoff value" in breath test and "reduction of A value less than 15% when comparing with pretreatment" in serology were made as the H. pylori positive criteria for monitoring therapeutic effectiveness on H. pylori. At the second stage of this study, total of another 63 of patients were studied to test the accuracy of the monitoring criteria. The results showed that the sensitivity of breath test was all 100.0% and specificity 95.5% and 100.0% respectively at 6 and 12 month after termination of treatment, the sensitivity of serology was all 100.0% and specificity was 50.0% and 83.3% respectively. A scheme for monitoring the effect of anti-H. pylori chemotherapy by combination of 14C-urea breath test and serology is proposed based on this study. PMID- 8731861 TI - [A multiple center clinical trial on fluconazole for deep-seated fungal infections]. AB - A multiple center clinical trial was conducted to evaluate the efficacy and safety of domestic fluconazole in treating 913 cases of deep-seated fungal infections. Fluconazole was given 100-200 mg daily for 3 days to 8 months. The results showed that the cure rate and the total efficacy rate were 69.26% and 94.29% respectively. The fungal clearance rate was 93.83%. The main side-effects were nausea, vomiting, diarrhea and abdominal pain, but most of the patients could endure. The side-effect rate was 9.20%. This clinical trial indicated that domestic fluconazole was effective and safe in treating deep-seated fungal infections. PMID- 8731862 TI - [Lupus nephritis in male adults, an analysis of the clinical and pathological features]. AB - Systemic lupus erythematosus is an uncommon disease in men. The clinical and pathological features and prognosis in 50 male patients with lupus nephritis (LN) and 50 age-matched female patients with LN were analysed. The age at which the disease began and the diagnosis was made was generally older in the male. The incidence of LN was higher in the male than in female (P < 0.01). The incidence of types IV and V LN was more common in the male than in female. Type II LN was not found in male patients. Proteinuria over 3.5g/24h and renal failure were more common in the male than in female (P < 0.05). The three most common clinical manifestations in male patients were irregular fever, skin rashes and painful joints. Rashes, hepatomegaly and neuropsychiatric dysfunction were more common in the male than in female. The patients were followed up one to twelve years. The rate of recovery and improvement was lower, but that of relapse and mortality higher in the male than in female. PMID- 8731864 TI - [Key points in tendon surgery]. PMID- 8731863 TI - [Tumor necrosis factor and clinical diseases]. PMID- 8731865 TI - [Synovial fluid on non-synovial tendon: experimental study]. AB - The non-synovial tendon was placed in the synovial cavity of rabbit knee joint to carry out tissue culture in vivo. The surface of the cultured tendon was covered with a smooth membrane-like tissue which possessed the morphologic characteristics of the synovial membrane identified by histological and electron microscopic examinations. The tendon segments were kept free in the cavity of knee joint and adhesions were not observed. It is suggested that non-synovial tendon could transform into a synovial tendon in the environment filled with synovial fluid. PMID- 8731866 TI - [Experimental study on transplantation inside and outside tendon sheath]. AB - The range of motion of PIP and morphological changes of tendon grafts were studied after inside outside tendon sheath grafting in rabbits. It was found that the inside tendon sheath grafting together with early postoperative mobilization healed without adhesion, and most of the cells remained viable. After outside tendon sheath grafting, the tendon cells proliferated with extensive adhesions. The range of PIP motion was greater after inside tendon sheath grafting than that after outside tendon sheath grafting. The results suggest that tendon grafting is donor-tissue-specific and the synovial layer of inside tendon sheath plays an important role in preventing adhesion formation. PMID- 8731867 TI - [Ultrastructural observation of tendonization of artificial tendon 109HH in rabbit]. AB - Ten New Zealand rabbits were divided into 5 groups at random. Calcaneal tendons were cut bilaterally, then atificial tendon 109HH was used to connect the two ends of the cut tendon. Ultrastructural changes of control group and experimental groups at 2, 6, 12, 28 weeks after section were observed. The results showed that fibroblast proliferated and a lot of ribosome and RER appeared in plasm during 2 approximately 6 weeks, indicating artificial tendon caused fibroblast proliferation. During this period, fibroblast over synthesized collagenous protein and the synthesis of collagenous fibers peaked. During 12-28 weeks, the number of fibroblasts and the synthesis of collagenous protein decreased. Finally, fibroblasts became inactive tendon cells. With the formation of new tendons, the artificial tendens were degradated and absorbed, and disappeared after 12 weeks. The new tendon fibers became thicker and had the correct direction through reconstruction. The structure and function of new tendons could be restored to be consistent with normal values. PMID- 8731868 TI - [Experimental research on using macromolecule sodium hyaluronate to prevent flexior tendon adhesion]. AB - With chicken paw as an animal model, we studied the effects of sodium hyaluronate product (SHP) on the heal of injured tendon and the formation of post-surgical tendon adhesions. The function of digit flexion angle and moving function of the tendon in SHP group were remarkably superior to those of tendons in physiological salt solution control group (P < 0.01). Tendons of the experiment group healed well, their surface was smooth without adhesion. False sheath was formed around the tendon, and there was also a gap between the false sheath and the tendon. However, the adhesion between the tendon and its surrounding tissue in the control group was donse with clear boundary. These results suggest that SHP has a protective effect against the formation of adhesion in healing tendon. PMID- 8731869 TI - [Experimental study of sodium hyaluronate products on prevention of tendon adhesion]. AB - In an experiment sodium hyaluronate product (SHP) was used to observe its effects on healing and preventing adhesion of rabbit muscle tendon. Several methods were used such as gross and histopathological observation, imaging analysis, anti tension strength measurement. The results confirmed that SHP could reduce collagen contents in peri-tendon tissues and fibroblasts after surgery and decrease the formation of scars in peri-tendon tissues. Therefore, it is effective in preventing muscle tendon adhesion and improving slide function of muscle tendon. Besides, it does not hinder muscle tendon healing. PMID- 8731870 TI - [Flexor tendon repair: timing of surgery and sheath management]. AB - Effect of timing of repair and management of sheath on functions of injuried tendons was evaluated in a clinical relevant model using 60 white leghorn chickens. The flexor digitorum profundus (FDP) tendons of the long toes were cut and were repaired at 1, 4, 8, 14 and 20 days after injury. The sheath on the left foot was closed and that on the right excised. The tendon excursion, toe motion, morphology of the sheath and tendon healing were studied 6 weeks after tendon repair. Tendon functions were the best in primary repair, and they got worse as the delay became longer. Repair in the late stage produced significantly worse tendon functions. Sheath closure with remarkable tension worsened tendon healing. We conclude that the optimal timing for tendon repair is at the primary stage. Delayed tendon repair can be performed within 3 weeks to one month after injury, but the repair is preferably done in initial several days of delay. Direct sheath closure is not advocated in tendon repair in the delayed period. PMID- 8731871 TI - [Transposition of flexor pollicis brevis muscle for reconstruction of opposition function of the thumb: anatomical study and clinical application]. AB - Anatomical study was performed on 20 cadaveric hands and the results showed that M. flexor pollicis brevis overlap the M. abductor pollicis brevis for about half of its width at their origins and the overlapping reduced to about 1/3 of the width at the muscle belly level. The M. flexor pollicis brevis chiefly inserts on the palmar aspect of the base of proximal phalanges. The M. abductor pollicis brevis primarily inserts to the radial side of MP joints, the M. flexor pollicis brevis is entirely innervated by the deep branch of the ulnar nerve. In order to increase the angle bettween longitudinal force lines of these two muscles for 7-9 degrees, the authors transfered the insertion of the M. flexor pollicis brevis to the radial side of the metacarpal-phalangeal joint, so that it gives this muscle the function of opposition. Eight patients had been treated and followed up on average for 12 months. All had fine functional results. This new methodis effective, least traumatizing and does not need transposition of another tendon. PMID- 8731872 TI - [Experimental research and clinical application of allogenic tendon grafting]. AB - Autologous free tendon grafting is a very common treatment for tendon defect of the hand. However, it is difficult to obtain autologous free tendon grafts to repair patients with multiple tendon defects. We described the treatment using deep-freeze-stored tendon with MEM and the establishment of clinical tendon bank. Allograft tendons were used successfully in 15 patients clinically. In the experiments, we also performed a series of studies on immunity and repairing ability of the allogenic tendon grafts. The results indcated that the treatmnet using deep-freeze-stored tendon with MEM could reduce the immunogenicity of allograft. There was no significant difference between autograft and deep-freeze stored allograft 24 weeks after operation, but there was a significant difference in the fresh allograft 3 weeks after grafting. Based on animal experiment, we established a clinical tendon bank. Deep-freeze-stored allogenic tendon grafts can be used successfully in place of autologous tenden grafts in treatment of tendon defect of the hand. PMID- 8731873 TI - [Influence of age on severe pancreatitis]. AB - Eighty-four patients of severe pancreatitis were divided by different line of age and the mortality of patients with severe pancreatitis above and below the line was comared. The results showed that the 60-year of age was the age with the lowest mortality. The analysis of 35 patients with severe pancreatitis who were older than 60 years of age indicated that multiple gallstones was the most common cause initiating the disease. The common severe concurrent diseases were hypertension, coronary disease, cerebrovascular disease, and respiratory disease. Common organ failures were ARDS, shock and heart failure. The number of failed organs in elderly patients in the fifth and seventh day after admission was obviously more than that in young patients. PMID- 8731874 TI - [An evaluation of nonsurgical treatment for severe acute pancreatitis]. AB - In this prospective study, the rationality of non-operative treatment for severe acute pancreatitis was evaluated. Twenty-four patients aged 15-74 years with a mean age of 46.1 +/- 15.5 years were divided randomly into two groups: 12 patients in nonoperative group and 12 in operative group. The patients treated nonoperatively survived, while 5 patients died of operation. It is suggested that patients with severe acute pancreatitis can be cured by nonoperative management. PMID- 8731875 TI - [The treatment of gastric carcinoma in III stage]. AB - From 1975 to 1989, we treated 238 cases of gastric carcinoma in III stage, 90.7% of these were followed up. The total 5-year survival rate was 45.4%, 10-year survival 33.7%. The 5-year survival rate and 10-year survival rate (56.3% and 47.5%) in the extensive radical treatment group (102 cases, selective R3 or R3) was much higher than that of routine radical treatment group (114 cases R2). The 5-year survival rate and 10-year survival rate in the group with auxillary post operational chemotherapy were much higher than those of the group without chemotherapy. The post-operative survival rate in the group with chemotherapy plus Chinese medicine was much higher than that of the group with chemotherapy alone. The peritoneal metastatic rate of the group with peritoneal flushing plus chemotherapy (18.7% and 20.6%) was much longer than that of the group without peritoneal flushing and/or chemotherapy (31.9%). We consider that the best treatment of gastric carcinoma in III stage is the sufficient removal of gastric lymph nodes for the prevention of metastasis through peritoneal and circulatory route. PMID- 8731876 TI - [Value of surgical treatment in patients with gastrointestinal metastatic tumors]. AB - A retrospective review was completed on 43 patients with metastatic lesions to the gastrointestinal tract between 1980 and 1993. The most commonly involved organs were stomach and small intestine. The main features were abdominal pain (70%), acute abdomen (42%), alimentary bleeding (21%), massive in 11.6%). Operations were performed on 42 patients (palliative resection in 40 patients, enteric bypass in 2). The results support surgical intervention for patients with complaints of gastrointestinal metastases, especially in emergency patients. PMID- 8731877 TI - [Detection of micrometastases in the bone marrow in patients with gastric cancer]. AB - An immunocytochemical method using an antibody probe to recognise the epithelial membrane antigen was used to screen smears obtained surgically from bone-marrow in 88 patients with gastric cancer. Tumor cells were detected in the bone-marrow of 58 patients (65.9%). The EMA positive cells in the marrow were not correlated with the location and node status of the stomach. In the stage of TNM I, II, III and IV, the positive rates of micrometastases in the bone marrow were 42.9%, 57.1%, 73.7% and 69.0%, respectively. The results showed that the poorer differentiated lesion, the higher rate of positive cells in the bone marrow. The curative surgery and multimodality treatment after operation could result in remission of positive cells in some patients. The method can detect occult metastases in bone marrow, and may be useful to monitor patients for evidence of response. It can measure the efficacy of adjuvant therapy, and predict prognosis of the patients. PMID- 8731878 TI - [Non-functioning adrenal tumor: clinical analysis of 29 cases]. AB - 29 patients with adrenal tumors were admitted to our hospital from June 1985 to July 1994. The diagnosis was made preoperatively in 7 patients (5 adrenal cyst, 2 adrenal myelolipoma) by B-US, CT or MRI. Of the 22 patients whose diagnosis was not made before operation, 8 (tumor diameter: < 3cm) had adrenal cortical adenomas, 6 (tumor diameter: 3-6cm) adrenal medullary tumor, 8 (tumor diameter: > 6cm) adrenocortical cancer (2), adrenal neuroblastoma (1), metastatic cancer (2), adrenal pheochromocytoma (1) and adrenal lymphatic hyperplasia (1) respectively. All non-functioning adrenal tumors should be treated operatively except for adrenal cyst smaller than 3 cm or adrenal myelolipoma. PMID- 8731879 TI - [Orthotopic heart transplantation: a case report]. AB - In July 1994, we performed orthotopic heart transplantation for a patient with hypertrophic myocardiopathy, who had had open heart surgery enlarging the right ventricular outlet. The patient had an excellent postoperative result. He received triple-drug immunosuppressive therapy. Large dose of methyl-prednisolone was used successfully for his acute rejection reaction. At present, the patient is living well with good cardiac, hepatic and renal function. We described donor heart harvest, myocardial protection, operative procedure, immune monitoring, anti-rejection and anti-infectious therapy. We introduced the process of protecting donor heart on the way to hospital by retrograde perfusion with medium hypothermic oxygenated-blood combined with St. Thomas cardioplegia. Besides endomyocardial biopsy, other noninvasive examinations such as noninvasive cardiac output monitoring could be helpful to monitor acute rejection reaction. PMID- 8731880 TI - [Surgical treatment of ascending aortic dissecting aneurysm with Cabrol's operation in 3 patients]. AB - Form Mar. 1993 to Dec. 1994, 3 patients with ascending aortic dissecting aneurysm underwent aneurysmal incision, artificial vessel interposition, aortic valvular replacement and coronary artery transposition as Cabrol's operation. All recovered health discussed. The procedures and techniques were modified on the Cabrol's. Anastomosis of coronary openings with 8 mm Gore-tex tube obtained good match, little tension, and avoided intimal tear around coronary ostia and blood leakage of the anastomosis. We have created a self blood infusion system to stop hemorrhage of vessel graft or anastomosis by a bypass between right atrium and aneurysmal sac around the dacron graft. PMID- 8731882 TI - [Shape memory angioembolus for varicocele]. AB - Seven target veins (in 5 dogs) were embolized with nitinol conical rugby embolus (NT-CRE). The pathological examination showed firm impact between NT-CRE and intravenous membrane without misembolization from retrograde flow. Compact collagenous fibrous tissue was seen in and out NT-CRE on spaces. On the basis of animal experiment, 10 patients with varicocele were treated with NT-CRE. After the operation, the varicocele and symptoms disappeared and venae spermatica collapsed. The follow-up for 1.5-2.3 years showed no recurrence in the 10 patients. We consider that the therapeutic method is simple and effective for varicocele. PMID- 8731881 TI - [Plasma prostacyclin (PGI2) levels in peripheral venous, arterial and portal venous blood in cirrhotic patients with portal hypertension and their clinical implication]. AB - Plasma levels of PGI2 were measured by radioimmunoassay in 20 consecutively admitted cirrhotic patients with portal hypertension from Aug. 1993 to Aug. 1994. It was found that cirrhotic patients with portal hypertension had much higher portal venous PGI2 levels than controls (528.25 +/- 205.48ng/L vs. 235.73 +/- 49.36ng/L, P < 0.001). Portal venous PGI2 levels in patients with cirrhosis and portal hypertension correlated significantly with portal venous pressure (r = 0.60, P < 0.05). The results indicate that increased portal venous PGI2 levels may have a role in maintenance of established portal hypertensive state in cirrhotic patients with portal hypertension. PMID- 8731883 TI - [Effects of thermal injury on production of nitric oxide in rat heart]. AB - To elucidate the relationship between thermal injury and production of nitric oxide (NO), we measured the levels of heart No2-/NO3- (the stable and products of NO) and GMP in rats with full thickness burn of 30% TBSA. A higher level of heart tissue NO2-/NO3- was observed from 3 to 24 hours after burn injury. Parallel to the changes of NO2-/NO3-, there were increases of tissue cGMP level and heart tissue water content in burned rats. L-NMMA, a specific inhibitor of NO synthetase (NOS), can block the rise of heart NO2-/NO3- and the enhancement of heart water content. The results indicate that the heart injury caused by burn injury may be associated with induction of NOS and formation of NO. PMID- 8731884 TI - [Effects of Chinese herbs on impaired lymphocyte functions after thermal injury in mice]. AB - Four Chinese herbs were chosen for this study. They were Panax pseudoginseng wall (PPW), Loniconica japonica thunb (LJT), Polygonum cuspidatum (PC) and Jilin ginseng (JG). They were given from the 2nd to the 7th postburn day (PBD). The results showed that the response to antigen signal, the proliferative capacity, the interleukin II production function, and the antibody production ability by lymphocytes in control burned mice were more noticeably reduced than that in the normal mice. The PPW, LJT, PC might restore all the impaired functions in different degrees. The JG showed no effect at all. PMID- 8731885 TI - [Percutaneous autologus bone marrow grafting in the treatment of ununited fractures and bone defects]. PMID- 8731886 TI - [Hoffa disease: diagnosis and arthroscopic treatment]. AB - The diagnosis of Hoffa disease (syndrome of impigement of the infrapatellar fat pad) was established in 20 patients with anterior knee pain by clinical physical examination, magnetic resonance imaging and arthroscopic findings. Arthroscopic partial or total resections of the infrapatellar fat pad were performed after arthroscopic examination. According to the Larson's rating score for knee function, 18 cases had a satisfactory result at an average follow-up of 36 months. The results of follow-up also suggested there was no significant difference between the partial and total resections of the fat pad. PMID- 8731887 TI - [Diagnosis and treatment of lateral crypta synovial crush syndrome of knee]. AB - Lateral crypta synovial crush syndrome is often confused with lateral meniscus injury. Thirteen cases were reported in this paper. Of them, eleven cases had pain at the lateral of their knees which were flexed from twenty degrees to eighty degrees. One patient had knee pain in erect position and other one on walking. Local tenderness at lateral tibial plateau of knee existed in all cases and small nodule could be palpated at tenderness point in ten cases. Ten cases were examined by arthroscope. Synovial neoplasm or abnormal synovial fold was discovered in lateral crypta of knee which could be pushed into joint space at flexed twenty degrees to eighty degrees. After removing them, symptom disappeared. Three cases were cured by nodule block therapy. The block therapy of injection of prednisolonum was available for not only diagnosis, but also treatment. The indication of arthroscope is that effect of block therapye is unsatisfactory. That all cases occurred on the lateral side might be related to the diffierence in anotomic structure. PMID- 8731888 TI - [Arthroscopic evaluation and treatment of the disorder resulted from open meniscectomy of the knee]. AB - Between 1989 and 1994, twenty four cases of knee disorders caused by open meniscectomy had undergone the second operation by arthroscopic procedures. The results of retrospective study indicated that main reasons of early disorders following meniscectomy were locking of remanent posterior horn, tears of remanent meniscus and injuries of fat pad and the cause of late disorders were single compartment osteoarthritis and genu varum/valgum caused by meniscectomy. In early cases, the arthroscopic reshape or removal of meniscal remanents and partial resection of fat pads could get satisfactory results. In late cases of osteoarthritis with genu varum/valgum, arthroscopic debridement, high osteotomy of tibia or knee replacement should be carried out as means of treatment. PMID- 8731889 TI - [Adjustable reconditioner for the treatment of patella fracture]. AB - Patella fracture belongs to the articular fracture. The operative methods to fix the patella fracture are numerous and there is no unanimous view. In order to protect the reconditioned patella, usually it is necessary to fix externally for a period of time. Since 1989, the authors have treated 165 cases of patella fracture with self designed, adjustable reconditioner and have acquired satisfactory results both in its fixing effect and function recovery. There was no need to fix the patella externally and the patients could do functional training in early stage. According to the assessment of the postoperative function and the motion limits of the knee and rehabilitation of the fracture, the results of the treatment were: excellent in 148 cases and good in 17 cases. This article describes the fixing mechanism of the reconditioner. It consists of the left and right petals and a screw. According to different sizes of patella and various types of the fracture, the reconditioner could be adjusted to connect the fracture closely with the petals and a screw. The structure of the reconditioner is simple, the operation is easy to carry out. It is adjustable during the operation and holds the fracture firmly and secure. It is suitable for all kinds of patella fracture avoiding partial or total resection of the patella. The other facture of the reconditioner is that the fixed patella could bear traction force when bending the knee at 90 degrees, so the patient could walk and squat on the heels normally. PMID- 8731890 TI - [The use of the Ho: YAG laser in arthroscopic surgery]. AB - From May 1993 to April 1994 we had performed 72 cases (total 84 joints) of arthroscopic operations with holmium laser. It included 12 bilateral knees, and single joints in the shoulder, elbow, wrist and ankle. We performed partial menisectomy, chondroplasy, partial synovectomy and lateral retinacular release. The energy used was from 1 to 12kJ depending the complexity of the operations. All these cases had been followed up 6 to 16 months. The function of the joints was good, only two cases had joint effusion. The rate of good and excellent results was 97%. PMID- 8731891 TI - [Simultaneous bilateral total knee replacement for the patients with serious rheumatoid disease]. AB - This retrospective study reviews 57 patients with serious rheumatoid disease who had bilateral total knee arthroplasties simultanneously between 1987 and 1994. The short follow-up results revealed that there was no increase in complications in the patients with simultaneous bilateral procedures, and nearly identical postoperative results to those observed among patients with single joint replacement. Advantage of simultaneous bilateral total arthroplasty include reduction in hospital cost, the need for less invasive surgical event and the ability to rehabilitate the patient symmetrically. PMID- 8731892 TI - [A morphological study of degenerative mechanism of articular cartilage by abnormal high stress]. AB - The articular cartilage from the right knee of 87 rats whose left calcaneal tendons had been transected was examined under transmission and scanning electron microscopes, and light microscope. Morphological observation showed a few small focal areas of cartilage surface disruption, pit and collagen fibrillation at 2 weeks after operation. After 1 month, many clusters of chondrocytes could be found, showing hyperactive synthetic and secretary activity. The cartilage degenerated progressively. The initial lesion in high stress side was at the matrix of cartilage, the collagen network was disrupted, then proteoglycan become degraded. The microenvironment of chondrocytes was changed with degradation of matrix. The chondrocytes became degenerative. The degeneration of chondrocytes influence the matrix again. A vicious cycle was thus formed. At the early stage of the degeneration of the cartilage induced by high-stress, som chondrocytes proliferate and have hyperactive synthetic and secretive functional activity. But soon, most of the chondrocytes degenerate. That is because of the chondrocytes have got repaired in response to initial matrix lesion and the normal microenvironment had degraded with aggravation of matrix lesion. These chondrocytes again became degenerative quickly. PMID- 8731893 TI - [Evaluation of surgical resection for gastric cancer extending to adjacent organs]. AB - From June 1972 to December 1993, 280 patients underwent surgical resection for their gastric cancer extending to adjacent organs. The patients were divided into three groups. Group A, 93 patients, were treated with gastrectomy plus complete removal of the invaded organs. Group B, 55 patients, underwent gastrectomy with incomplete removal of the invaded organs. Group C, 132 patients underwent gastrectomy only. The 5 year survival rate of each group was 32.7%, 7.7% and 4.4%, respectively. The 5 year survival rate in the group A was significantly higher than that in two other 2 groups (P < 0.05). The 5 year survival rate in the group B and C was similar (P > 0.05). The patients without incurable factors (liver metastasis, peritoneal dissemination and widespread nodal involvement) underwent gastrectomy with complete excision of invaded organs, irrespective of the number or site of invaded organs. On the other hand, the patients with incurable factors should be given palliative gastrectomy only. Because symptoms are alleviated, the quality of life is improved and the survival time is prolonged. PMID- 8731894 TI - [Conservative surgery and radiation therapy in the treatment of operable breast cancer]. AB - Forty-five patients with breast cancer were treated in our hospital by conservative surgery and radiation therapy from 1985 to 1994. There were 22 patients in stage I and 23 in stage II. All the patients were received breast preserved operation including segmental resection and axillary node dissection. Routine postoperative irradiation was given to the patients with or without regional lymphatic melastasis. The indications of chemotherapy and endocrinotherapy were as same as those of Halsted's radical mastectomy. The duration of following-up was 3-5 years. The 5-year local recurrence was 8.8%. The 5-year OS was 94.1%, and DFS 94.1%. PMID- 8731895 TI - [Significance of diagnosis and prognosis for breast carcinoma with examination of Ag-NORs]. AB - The argyrophilic nucleolar organizer regions (Ag-NORs) was measured in the tissue sample aspirated with fine needle for 17 cases of breast benign lesion and 46 cases of breast cancer, by analysis of their micro-image with light microscope and computed. The results indicated that the amount of Ag-NORs in the nuclei of breast cancer was increased while the percentage of average areas occupied by Ag NORs was decreased. It was significant statistically in comparing with benign lesion (P < 0.05). In addition, the amount of Ag-NORs in the nuclei of breast cancer with and that without metastasis was not different markedly (P > 0.05). However the percentage of average areas occupied by Ag-NORs was different statistically (P < 0.05). The conclusion of Ag-NORs is of benefit to distinguish from benign and malignant tumours of breast as well as significant for evaluating the prognosis for breast cancer. PMID- 8731896 TI - [Hemodynamic study of distal splenorenal shunt (DSRS)]. AB - Color-flow Doppler sonography (FDS) was used for assessing the hemodynamic changes in 8 patients undergoing distal splenorenal shunt (DSRS). Results were compared with 14 patients with portal-azygos disconnection (PAD). It was found that 3 weeks after DSRS portal venous flow (PVF) decreased by 50%, and at least in some patients the reduction of the PVF was a continuous process during the period of two years in the DSRS group. The postoperative decrease in PVF in DSRS patients was in sharp contrast to that in PAD patients (P < 0.001), hence PAD is superior to DSRS in remaining satisfactory PVF. PMID- 8731897 TI - [Potassium metabolism in warm heart surgery]. AB - The dynamic balance of potassium getting in and out the body during and after operation was surveyed in 26 adult patients who underwent cardiac surgery with normothermic cardiopulmonary bypass (CPB). We found that during CPB, because of the continous perfusion of warm oxygenated blood cardioplegia solution, a large volume of potassium entered the body; meanwhile, only a little potassium removed from the body in urine. At the moment of CPB finish, the volume of residual potassium in the body was as large as 74.5mmol (mean), that was 1.2mmol/kg body weight. The mean plasma potassium level at the points of aorta cross clamp release and CPB finish were 6.8mmol/L and 5.5mmol/L respectively, and all in normal range at several times measured from the end of operation (usually 12-2pm) to the second postoperative day 7am. The volume of potassium replenishment according to urine output in this group was much less than those in hypothermic CPB patients from the end of operation to the first postoperative day 7am. During CPB extracellular potassium shifted into intracellular fluid, but the shifting became opposite around the operative day 6pm, and the final equilibrium of potassium metabolism was achieved at the second postoperative day 7am. The remarkable hyperkalemia at the point of aorta cross clamp release may delay the heart rebeating ( > 5min, 35%), but the automatic rebeating rate was still very high (96%). With regard to the prevention of hyperkalemia during CPB, we also give some suggestions in this paper. PMID- 8731898 TI - [Diagnosis and management on acute coronary artery occlusion in valve replacement surgery: a report of four cases]. AB - This paper reported four cases of acute coronary artery occlusion occurred during valve replacement procedure. Case 1 was of Marfan's syndrome who underwent Cabrol procedure. Case 2 was of congenital aortic bileaflet malformation with severe regurgitation and simple right coronary artery, who received the procedure of AVR. Both cases 3 and 4 suffered from rheumatic valve disease who were given DVR and tricuspid valve annuloplasty. All the cases had acute left coronary artery occlusion and did not revive after removal of aortic cross-clamp, which were immediately corrected by coronary bapass grafts from aortic to left descending branch with saphonous veins. All the patients successfully weaned from asistant circulation. Two cases survived. The causes of acute coronary artery occlusion were analyzed and the diagnostic methods and management introduced. PMID- 8731899 TI - [Adrenocortical carcinoma: report of 20 cases]. AB - The authors reported 20 cases of adrenocortical corcinoma. The tumors were nonfunctional in 12 cases (average age 49.7) and functional in 8 cases (5 with hypercortisolism, 3 with adrenogenital syndrome; average age 8.8). The prognosis in this group were poor, the survival of one year was 40%, two years 35% and five years 10%. Most of adrenocortical carcinomas in childhood were functional and their prognosis were poor because of delayed treatment. The nonfunctional tumors were usually diagnosed in the late period, therefore the prognosis was worse, but in recent years some cases of no secretory carcinomas had been occasionally found by image diagnosis and these tumors were small, so their prognosis was favorate. PMID- 8731900 TI - [Experimental study of implantation of autologous vascular endothelial cells on dacron grafts]. AB - Ten mongrel dogs underwent carotid artery bypass 17 times with dacron grafts, 4mm in inner diameter and 5cm in length. The grafts in experimental group A were immediately seeded with autologous endothelial cells harvested by 0.25% trypsin solution (n = 7). The grafts in experimental group B were seeded with autologous endothelial cells cryopreserved in liquid nitrogen for 1 week (n = 3). The grafts in contrast group was not seeded with any endothelial cells implanted in the contralateral carotid arteries in group A (n = 7). The number of endothelial cells harvested in the two groups was 0.9 +/- 0.3 x 10. Factor VIII related antigen stain method confirmed the cells harvested by 0.25% trypsin solution to be endothelial cells. The grafts were removed and studied at the end of 2, 4, and 6 week after implantation. The total patency rate of group A was 85.7% (6/7), and that of the contrast group 57% (4/7). The weight of thrombus in group A was 40.6 +/- 36.9mg and the contrast group 85.9 +/- 26.3mg (P < 0.01). Scanning electron microscopy revealed that the cells at the middle segment of the grafts in the group A and B had the characteristics of ellipse, and tight intercellular junction, while the middle segment grafts in contrast group were covered by platelets, erythrocytes, leukocytes and fibrin. Transmission electron microscopy, factor VIII related antigen stain method, and vimentin stain method demonstrated that the cells in the group A and B showed characteristics of endothelial cells. It was concluded that endothelialization of the dacron grafts could be accelerated and the quality could be improved by immediate seeding with endothelial cells or by seeding with endothelial cells cryopreserved in liquid nitrogen for 1 week. PMID- 8731901 TI - [A stereologic study on the influence of stent tube pressure on bilio-intestinal anastomotic scar formation]. AB - Biliary tract injury results often in biliary scar stricture causing obstructive jaundice and biliary cirrhosis. Choledocho-jejunostomy with tube stent is the routine to prevent anastomotic stenosis, but how long the stent tube should be in place is controversial and little is known on the influence of the stent tube on anastomotic scar formation. In canine experimental models, bili-intestinal scar was sampled 3 weeks, 3, 6, 9 and 12 months after anastomosis for ultrastructure stereologic image analysis. It was found that the internal pressure of the stent tube exerting on anastomotic stoma inhibits scar formation, and the stent tube should be remained inside the stoma for not less than 9 months. PMID- 8731902 TI - [A quantitative study on alpha-1 adrenergic receptor in the liver of Wistar rats during the process of CCl4-induced hepatic cirrhosis]. AB - In this study, radioligand binding analysis was used to quantitatively measure the concentration and affinity of alpha-1 adrenergic receptor in the liver of Wistar rats during the process of CCl4-induced liver cirrhosis. It was found that compared with normal controls, the maximal binding capacity (Bmax) of alpha-1 adrenergic receptor in the liver of 13 pre-cirrhotic rats decreased from 132.8 +/ 25.0fmol/mg to 96.9 +/-20.8fmol/mg of protein (P < 0.01) with increased dissociation constant (Kd, from 0.1633 +/- 0.0322nM to 0.3079 +/- 0.0786nM, P < 0.01). The Bmax in 20 cirrhotic rats decreased significantly from 132.8 +/- 25.0fmol/mg to 71.9 +/- 17.7fmol/mg of protein (P < 0.01), while Kd values increased from 0.1633 +/- 0.0322nM to 0.4012 +/- 0.0965nM (P < 0.01). These findings indicate that the decrease of binding capacity and affinity of alpha adrenergic receptor may play a role in the disturbances of metabolism of catecholamine often seen in some cirrhotic patients and have implications in the pathogenesis of portal hypertension. PMID- 8731903 TI - [Experimental study of survival mechanism of skin flap with subdermal vascular rete]. AB - AIM: To determine the blood supply style and survival mechanism of skin flap with subdermal vascular rete. MATERIAL AND METHODS: The skin flap on trunk of piglets were taken as experimental pattern. The survival length of skin flap was examined and the nuclear element distribution in the skin flap was determined by ECT. RESULTS AND CONCLUSION: This study demonstrated that the distal portion of super long flap with subdermal vascular rete fail to getting blood supply from its pedicle. This kind of flap is actually a combination of flap and free graft with subdermal vascular rete. This kind of super-long and super-wide flap is actually not a flap but a free skin graft. The survival of super-long and super-wide flaps with subdermal vascular rete rely on the blood supply from the wounds and their edges partially or completely. Their survival style is actually free grafting. PMID- 8731904 TI - [An experimental study of the expression and localization of tumor necrosis factor mRNA in liver of rats with severe burn]. AB - In situ hybridization was used to study the expression and localization of tumor necrosis factor (TNF) mRNA in liver of rats after burn. The results showed that the Kupffer cells expressed TNF mRNA in normal rats. After burn, the number of positive expression cells increased quickly, the expression reached max-mum at 6h postburn, and there were no significantly different with normal at 24h postburn. The dynamic changes of number of TNF mRNA producing cells was similar to that of the portal plasma endotoxin level. Kupffer cells located in sinusoid surrounding portal vein expressed TNF mRNA especially at 6h-12h postburn. Also the sinusoid endothlial cells and infiltrative inflammation cells surrounding portal vein were the TNF mRNA-producing cells, this changes appeared significantly at 12-24h postburn. The results suggested that the endotoxin originated from gut may be the principal stimulator for expression of TNF mRNA. The expression of endothlial cells and neutrophils suggested that these cells had been activated. So the endothlium, neutrophils and Kupffer cell play an important role in the pathogenesis of the endotoxin-induced live injury. PMID- 8731905 TI - [Cryosurgical resection of pulmonary metastases (experience of twenty years)]. AB - Cryosurgical resection of pulmonary metastases was performed in 112 patients from 1973 to 1993. The cumulative 1, 3, 5, 10 year survival rate was 80.37%, 41.81%, 28.01% and 17.28%. One patient died of respiratory failure after operation. Thirty-day mortality rate was 0.89%. The disease-free interval (DFI) between the control of primary tumor and appearance of metastases was significantly correlated with post-thoracotomy survival. The multiple factor analysis also demonstrated that DFI was the most important prognostic factor for patients after cryosurgical resection of pulmonary metastatic lesions. The overall 5 year survival rate of patients treated with cryosurgery in our report was higher than that of those treated by local resection of pulmonary metastases reported by other authors. Our results indicated that cryosurgical resection of pulmonary metastases was effective and safe, the survival time of most patients could be prolonged and some patients could be cured. PMID- 8731906 TI - [Iatrogenic bile duct injuries during the process of laparoscopic cholecystectomy]. AB - Twelve patients with iatrogenic bile duct injuries occurred during laparoscopic cholecystectomy (LC) were treated from June 1992 to May 1994. All the patients underwent re-operation and were cured. The causes and characteristics of the injuries were: (1) perforation of the common hepatic or common bile duct caused by dissecting hook (3 cases); (2) necrosis and perforation of the common hepatic duct due to diathermic injury (1 case); (3) clamping of the common hepatic duct by Ti clip (1 case); (4) secondary high bile duct stricture following a failed end-to-end anastomosis or hepatico-cholangio-jejunostomy of the amputated common hepatic duct (5 cases); (5) delayed high bile duct stricture (2 cases). It is emphasized that the severity of bile duct injuries by LC be should not overlooked, and more experience in this field be accumulated to avoid this serious complication. PMID- 8731907 TI - [Surgical treatment for iatrogenic damage at the confluence of hepatic duct]. AB - Since 1989, 4 patients with iatrogenic bile duct injuries at the level of bifurcation were treated. Among them, the bifurcation together with both right and left hepatic duct had had wrongly been resected in 2 cases. The bifurcation was partially resected in the other 2 cases. The establishment of a permanent hepaticoentertostomy is essential. Abnormal bile flow would impede surgical effect, hence in the author's opinion, peripheral hepaticoenterostomy such as Longmire's procedure is unsuitable for iatrogenic bile duct injuries. The author's special technique for adequate exposure and accurate dissection of intrahepatic bile ducts is described. Four cases have been followed up for 7 months to 5 years with good result. PMID- 8731908 TI - [The trend of the gallstone disease in China over the past decade]. AB - The present study is the second national survey on 3911 srugically treated cases of gallstones which was collected from 33 hospitals of 5 provinces and 2 main cities of China and aims at reevaluating and comparing the trend of the gallstone disease in China since the first national survey was completed 10 years ago. The results showed that cholelithiasis remained one of the commonest surgical diseases and accounted for 11.5% of overall hospitalized patients during the same period of time and it occurred mostly in females of over 50 years of age, with a female-male ratio of 2.57 Vs 1.80% of the patients with single positioned gallstones had their stones in gallbladder and was mainly cholesterol stone in nature while pigmented bile duct stones was found only in 10% of the patients, which made the gallbladder Vs bile duct stone ratio and the cholesterol Vs pigment stone ratio a significant difference as 7.36 : 1 and 3.4 : 1 respectively when compared with the results of 1.5 : 1 and 3.4 : 1 from the first national survey, 10 years ago. A significantly decreased incidence of biliary ascariasis and biliary bacterial infection rate as well as the dietary changes from low protein, fat high vegetable to the opposite were directly relevant to the above alterations on the location and the type of the gallstones. PMID- 8731909 TI - [Free fatty acids in gallbladder bile]. AB - Using gas chromatography and high performance liquid chromatography (HILC), we examined free fatty acid and lecithin molecular species in gallbladder biles from patients with cholesterol gallstones. Effect of free fatty acids on cholesterol nucleation in model bile was studied by a sensitive cholesterol crystal growth assay. Compared to bile of controls, biles from patients with gallstones had higher total free fatty acid level, more palmitic acid, more stearic acid, more linoleic acid and arachidonic acid. The lecithin pattern was similar in all. After free fatty acids were added to model bile, palmitic acid, oleic, acid, linoleic acid and arachidonic acid had significant effect of pro-nucleating, free fatty acids on non-protein pro-nucleating factor. These data suggest that variations in quantitation and composition of free fatty acids are importanct in the pathogenesis of cholesterol gallstone formation. PMID- 8731910 TI - [An observation on nucleating activity of monoconjugated bilirubin (MCB) in model bile system]. AB - Recently, the increasing data suggest that the pathogenesis of cholesterol gallstones may be related to the imbalance between pro- and anti-mucleating factors in bile. Our previous results have shown that monoconjugated bilirubin (MCB) possessed certain pro-nucleating activity and might play an important role in the formation of gallstones. The present study was made to observe the effect of MCB on nucleating activity by using our previous established methods of purification and preparation of MCB in model system of supersaturated bile according to Kibe's method with cholesterol saturation index of 1.2, total lipid concentration of 10g/dl, and bile salt phospholipid ratio of 2.3. Nucleation time (NT) was observed in 3 experimental groups with six samples each and 2 different contents of lyophilized MCB was added to above model bile rendered a final MCB concentration of (1) 173 microM; (MCB 1 group) (2) 340 microM; (MCB-2 group) and (3) no MCB in the as control group. The results were NT 3.33 +/- 0.52, 2.17 +/- 0.41 and 5.83 +/- 0.75 days in groups. 1, 2 and 3 respectively with the P value < 0.05 when compared between the 2 groups. The nucleating activity (NA) value was 0.5712 and 0.3722 in groups 1 and 2, which revealed that the NA elevated in parallele with the dose of MCB added in model bile. Our findings suggust that MCB has a strong pro-nucleating activity in model supersaturated biles, and MCB may participate in the formation of cholesterol gallstones throngh its special pro nucleating activity. PMID- 8731911 TI - [The diagnosis and surgical management of parathyroid cysts]. AB - Six cases of parathyroid cysts were treated in our hospital during a ten--year period with an incidence of 0.27% of the total patients operated on for neck mass. Four females and two males had an average age of 47.5 years with a history of the disease from 3 monts to 30 years. All patients had a single cyst ranging in size from 2.7 to 7.0 cm in diameter, arising in the region of inferior parathyroid gladms. Five caese of asymptomatic neck mass were nonfunctioning cysts, which where thin--walled and contained clear watery fluid. One case associated with hyperparathyroidism was classified as functioning cyst confirmed histologically as parathyroid adenoma with intracapsular hemorrhage. Surgical excision was made in all cases and postoperative recovery was uneventful. The importance of imaging studies, percutaneous biopsy, and cyst fluid assay for PTH in preoperative diagnosis was emphasized. PMID- 8731912 TI - [Preoperative embolization of paraganglioma in head and neck]. AB - Preoperative embolization of 10 cases of paraganglioma in carotid artery body and jugular foramen region via endovascular approach was performed. Tumor vessels were obliterated 100% in 7 cases and 80% - 90% in 3 cases as was demonstrated by angiography. It is available for us to reduce bleeding, expose tumor, avoid the iatrogenic damage of the important vessels and nerves around tumor during operation and to total resection possible. Additionally, the manipulating points of embolization are discussed. PMID- 8731913 TI - [The indication and technique in total correction of tetralogy of Fallot: experiences in 212 patients]. AB - Two hundred and twelve consecutive patients underwent total correction of tetralogy of Fallot at Fuwai Hospital in Beijing between Sept. 1987 to Dec. 1993. They were 131 males and 81 females. Their age ranged from 1 year and 6 months to 37 years. 53 patients were more than 16 years old. Most of them had a typical symptoms and manifestations like cyanosis and clubbing fingers etc. Their HB ranged 155 to 325g/L (mean 21.2 +/- 3g/L). It was more thant 180g/L in 134 patients. SBE was complicated in one patient and reoperation was performed in 3 patients. Maldevelopment or hypoplasia of pulmonary artery and left ventricle were found in 52 patients. Only 2 patients died from nonsurgical cause postoperatively. The mortality was 0.9%. The remaining patients recovered uneventfully. The results suggested that hypoplastic pulmonary artery and left ventricle were not a absolute contraindication of total correction of tetralogy of Fallot. The high hematocrit was also not a high risk factor for death. The key was to correct the pathology completely and manage the complications properly. PMID- 8731914 TI - [Evaluation of surgical treatment for patients with multiple aldosteronoma: analysis of 11 cases]. AB - From 1957 to 1994, 332 patients underwent surgical treatment for aldosteronoma at Ruijin Hospital. Among them, 11 patients (3.9%) were diagnosed as having multiple aldosteronoma. We ususally used the same method to make the qualitative diagnosis for both single and multiple aldosteronoma. In our experience, however, it was difficult to distinguish the unilateral multiple aldosteronoma by rutine B-type ultrasonography, CT or gamma-scintigraphy before surgery. The correct diagnosis could be obtained only by carefully exploring during the adrenal operation and a satisfactory prognosis also can be acquired if multiple aldosteronoma could be removed completely. PMID- 8731915 TI - [The study of p53 gene mutation in human bladder cancer]. AB - Alteration of p53 gene and the molecular biological difference between superficial and invasive bladder cancer were investigated by polymerase chain reaction-single strand conformation polymorphism analysis and direct sequencing technique. p53 gene mutations were examined in exon region 5 to 8 in 21 bladder cancers. Only one of 14 superficial bladder cancers was found to have p53 gene mutation. Four of 7 invasive bladder cancers with T2, T3 and T4 were found to have p53 gene mutation. p53 gene mutation was found none of the 4 cancers with grade 1, only one of cancers with grade 2, four of 12 cancers with grade 3. These results suggest that p53 gene play an important role in the development of human bladder cancer. PMID- 8731916 TI - [Plasma nitric oxide in patients with renal transplants]. AB - The plasma nitric oxide (NO) in 32 patients with renal allografts was observed before transplantation and in the early period of three months after transplantation. Endothelin (ET), cyclosporin A blood concentration, Creatinine (Cr), blood urea nitrogen (BUN) were measured simultaneously. The results showed that plasma NO level (24.01 +/- 1.15 mumol/L) before transplantation was significantly higher than that after transplantation. There was significant correlation between plasma NO level and ET, Cr, BUN and blood pressure. We consider that plasma NO concentration may reflect the function of renal transplant and play an important role in early renal injury after kidney transplantation. PMID- 8731917 TI - [Image and clinical correlative studies on cervical spinal canal stenosis]. AB - Clinical finding, X-ray and MR imagings in 47 patients who underwent operation for degenerative cervical stenosis were correalatively studied. JOA score was used for evaluation and it varied from 6 to 14 points, with an average of 10.13 points. All Torg ratios were smaller than normal value and that of C4 was the smallest. The reserve space, sagittal diameter of spinal cord decreased, especially in the diseased segments. Sagittal diameter of spinal cord showed on T2 weighted imaging and Torg ratio were in good correlation with JOA score (P < 0.01, r = 0.4815, r = 0.0993), MRI being superior to X-ray (P < 0.01). The MRI characteristic changes of degenerative cervical stenosis were subarachnoid space decrease or vanish, compression deformation of spinal cord and abnormal signal in spinal cord. MRI classification was established according to the segments involved, compression direction. compression degree and abnormal changes in spinal cord, can indicate clinical changes and guide diagnosis and operation. PMID- 8731918 TI - [Experimental study of Colles' fracture combined with dorsal radio-carpal subluxation]. AB - By cutting different ligaments and the distal end of radius of 20 fresh specimens with radiophotographic records, the author explored the mechanism of formation and traumatic anatomy of Colles' fracture with dorsal radio-carpal subluxation (DRSA). The results of experimental studies indicate that under normal condition, the change of radiolunate angle has some influence on the displacement of central point of lunate (CPL) but does not lead to DRS. The first is the rupture of the dorsal and palmar ligaments of radiocarpal joint and the second is the fracture of distal end of radius with a reversed palmar tilting angle of articular surface of distal radius. To treat the patients of Colles' fracture with DRS the previous palmar tilting angle should be restored to the full, as well as the anatomic reduction. The fixatiion in neutral or light dorsal extension position should be recommended. PMID- 8731919 TI - [An experimental study of the effects of stress-relaxation plate on bone remodeling]. AB - An ideal bone plate would provide rigid fixation to ensure stabilization of bone fragments at the early stage after fracture, while at the late stage osteoporosis induced by stress shielding effect of the plate should be prevented. However, there is no report of such a plate that could meet with all these requirements. Twenty-eight adult New Zealand rabbits were used in this experiment. Ordinary stainless-steel plate (no washer plate) and similar plate padded with ultrahigh molecular polyethylene washers in its screw holes (washer plate) were fixed respectively on mid-shelf of each side of tibiae. The animals were killed at 2, 4, 8, 12, 16, 20 and 24 weeks after operation, and the fixed bone segments of both tibiae were removed for light microscopy, polarized light microscopy and fluoroscopy. The results showed that the tibiae fixed with plate fixation both appeared bone resorption, but there was more severe resorption in bone with no washer plate fixation than that with washer plate fixation. Cancelization of cortical bone was seen in the former but not in the latter at 12 weeks postopertively. This demonstrated that the washer might creep and damage with fixation time, resulting in gradual decrease in the rigidity of the plate-screw system and thus reducing bone resorption as caused by stress shielding. PMID- 8731920 TI - [Experimental study on the arteriovenous reversal for revascularization of ischemic brain]. AB - Twenty-eight New Zealand white rabbits were randomly divided into experimental group (n = 20) and control group (n = 8). In the experimental group, the bilateral common carotid arteries were ligated, the blodd flow of right common carotid artery was reversed into the right cephalic internal jugular vein distal to the facial common vein, and the arterial and venous blood were obtained from the cerebral circulation pre and postoperatively for blood gas analysis. While in the control group, only the bilateral common carotid arteries were ligated. Both experimental group and control group were further divided equally into 4 subgroups for histological study at the 48th hour and on the 14th day after operation, cerebral angiography and ink injection on the 14th day of postoperation, and long survival observation, respectively. The results showed that all the animals survived. In the experimental group, the blood gas analysis revealed no statistical difference (P > 0.05) between pre- and postoperative. the cerebral circulation was rich and patent, and the cerebral cells were normal. In the control group, however, the cerebral blood flow was inadequate and the cerebral cells necrosed extensively. It is concluded that the adequate revascularization of ischemic brain can be created by the arteriovenous reversal. PMID- 8731921 TI - NisP is related to nisin precursor processing and possibly to immunity in Lactococcus lactis. AB - In this study, a plasmid was integrated into nisP, creating the first defined mutation in a nisin biosynthetic gene. The mutant strain secreted fully modified nisin with the N-terminal leader still attached. The presence of the leader was confirmed by N-terminal sequencing of the purified precursor. The dehydration and lanthionine formation of the precursor were already completed as active nisin could be formed by cleaving the leader from the inactive precursor by a trypsin treatment or by incubation with wild type cells. Nisin immunity of the NisP mutant strain was lowered to about 10% of the wild type immunity. The results show that NisP is needed fro precursor processing and for development of high immunity of nisin. PMID- 8731922 TI - Rapid isolation of large amount of plasma VLDL and LDL by a two step ultracentrifugation. AB - A rapid method to obtain large amount of VLDL and LDL by ultracentrifugation is described. The mixture of VLDL and LDL was isolated and concentrated from plasma by an ultracentrifugation at 265 000 g for 2 h. VLDL and LDL were separated and purified by a further ultracentrifugation at 265 000 g for 3 h. This method combines the advantages of both sequential flotation ultracentrifugation and density gradient ultracentrifugation. It can process a large volume of plasma in a short time. The purity of isolated VLDL and LDL was confirmed by the lipoprotein electrophoresis on agarose gel and PAGE and by the apolipoprotein electrophoresis on SDS-PAGE. This rapid economical method is of great value in practical application. PMID- 8731923 TI - [Study of HCV infection and viremia in patients with posttransfusion hepatitis in China]. AB - One hundred and sixty-nine patients with posttransfusion hepatitis (PTH) were investigated for etiology with serological and PCR methods. Our result showed that HCV was the major causative agent of PTH in this study. A part of patients (24/169) were also HBV markers positive. Three patients were negative for all detected viral markers, looking forwards to progressive study. Anti-HCV was detectable from 7 days to one year after onsetting of clinical symptoms, average time 54.62 days. HCV RNA could be detected in early time of HCV infection ( 6 to 20 days later), average time 8.72 days. A follow-up study of 84 patients showed that all patients remained anti-HCV positive during the observation for 1.5 -- 3 years, but their HCV viremia had three forms: transient, persistent or intermittent. The interaction between HCV and HBV by their superinfection was unknown, awaiting further study. PMID- 8731924 TI - An experimental study of the mechanism of andrographis paniculata nees (APN) in alleviating the Ca(2+)-overloading in the process of myocardial ischemic reperfusion. AB - The aim of this experiment is to study the mechanism of APN in alleviating the Ca(2+)-overloading in dog model during the process of ischemic reperfusion. In comparison with the sustained ischemic group, the parameters in the ischemic reperfusion group demonstrated: Ca(2+) of ischemic region of myocardial cell increased (P < 0.05), Na+ increased remarkably (P < 0.01), the activity of Ca(2+) ATPase dropped remarkably (P < 0.01), and MDA increased significantly (P < 0.01). Whereas in the group pretreated with APN, the Ca(2+) in the relevant area reduced (P < 0.05), Na+ decreased significantly (P < 0.01), the activity of Ca(2+)-ATPase and Na+-K+ ATPase increased remarkably (P < 0.01), and MDA decreased significantly (P < 0.01). These findings indicate tha APN may improve the activity of sarcolemma ATPase in alleviating the Ca(2+) and Na+ -overloading by decreasing the harmful effect of oxygen free radicals. PMID- 8731925 TI - Effect of captopril on membrane currents of ventricular myocytes. AB - Using whole-cell patch-clamp technique, the present study examined the effects of captopril on membrane currents of ventricular myocytes. The results were as follow: when the concentration of captopril was 6 x 10(-4)mol, the fast inward sodium current and slow inward calcium current were all decreased significantly (P < 0.05). Besides, when the concentration exceeds 10(-3)mol, the delayed rectifier potassium current was increased (P < 0.05) as well. But if the experimental concentration was under 6 x 10(-4)mol, these three kinds of currents was not affected. These findings suggested that captopril has no direct antiarrhythmic effect at the average therapeutic serum level. PMID- 8731926 TI - Effects of warm blood cardioplegic solution on myocardial protection. AB - To evaluate the effects of warm blood cardioplegic solution on myocardial protection, normothermic induction and terminal perfusion of oxygenated blood cardioplegia in combination with intermittent administration of cold blood cardioplegia during ischemia were studied in an isolated working rat heart model. The experimental protocol consisted of a 120 min cardioplegic arrest followed by 45 min normothermic reperfusion. Myocardial content of adenosine triphosphate (ATP), recovery of the left ventricular function, release of creatine phosphokinase (CPK) and ultrastructure of myocardium were assessed before and after ischemia. The results showed that the hearts preserved with warm blood cardioplegic induction and terminal perfusion had significantly higher levels of ATP, better recovery of cardiac function and lower releases and lower releases of CPK than those receiving cold blood cardioplegia alone, with myocardial tissue being of generally normal structure. These findings suggest that warm induction and terminal perfusion of blood cardioplegic solution can accelerate myocardial metabolic and functional recovery, preserve high-energy phosphate, reduce myocardial injury and enhance myocardial protection. PMID- 8731927 TI - Effect of captopril on antithrombus function of endothelium. AB - The experimental models of cultured porcine aortic endothelial cells (EC) in vitro were established. 6-keto-prostaglandin F1 alpha (6-keto-PGF1 alpha), thromboxane B2(TXB2)content, plasminogen activator (PA), plasminogen activator inhibitor (PAI) activity in cultured medium and cyclic adenosine monophosphate (cAMP) level in EC were measured with radioimmunoassay (RIA), chromogenic substrates methods, in order to assess the effect and mechanism of captopril (CP) on antithrombus function of EC. The results showed that after administration of CP, the contents of 6-keto-PGF1 alpha and cAMP and PA activity were significantly higher, PAI activity were remarkably lower than those of control group. These effects were dose-dependent. Our finding indicated that CP might act as a prospective drug for antithrombosis through promoting anticoagulation and fibrinolysis function and increasing antithrombus action of EC. PMID- 8731928 TI - Separation and expansion of tumor infiltrating lymphocytes of digestive system in vitro and their cytotoxicity. AB - Primary tumor tissues in the digestive system were harvested from 15 patients. By mincing, enzymatic digestion and gradient density separation, sufficient TILs (> x 10(6)) were obtained from 13 of 15 (88.7%) patients in vitro in the presence of 500 mu/ml of recombinant interleukin-2 and 5% fetal calf serum after one month culture. 92.3% (12/13) of TILs proliferated well in vitro (92.3%). TILs expanded from 10(/)-folds to 10(3)-folds after being cultured for one month. CD25+ cell of the most fresh TILs was more than that of peripheral blood lymphocytes. CD25+ cells of TILs during 4th week of the culture was significantly greater (P < 0.01) than that of fresh TILs. CD4+/CD8+ ratio was decreased during four culture weeks because of increase of CD8 cells. By using modified colorimetric MTT assay for measuring activity of TILs against various tumor cells the results showed that cytotoxicity of gastro-intestinal TILs in autologous tumor cells is greater than on the other tumor cells. PMID- 8731929 TI - Chronic pulmonary infection caused by Mycoplasma pneumoniae leading to pulmonary arteriole remodeling and pulmonary hypertension in rats. AB - The pulmonary arteriole remodeling in Wistar rats with respiratory infection induced by mycoplasma pneumoniae was observed using light microscopy and morphometry. The pulmonary artery pressure (PAP) and index of right ventricular hypertrophy (RVHI) were measured. The intimal and medial hypertrophy can be seen in the pulmonary arterioles, leading to vessel wall thickening and narrowing of the lumina. The total number of the pulmonary arterioles decreased (P < 0.01), and both pulmonary hypertension (Ppa 4.11 +/- 0.19 kPa) and right ventricular hypertrophy (RVHI = 34.96 +/- 3.91%) occurred. In addition, an interstitial pulmonary fibrosis (IPF) was found, in which the content of collagen in the lung tissue changed, i. e., type I collagen increased whereas type III one decreased, and the ratio of type I collagen to type III one increased. It suggested that respiratory infection induced by repeated MP may result in remodeling of pulmonary arterioles and are closely related to pulmonary hypertension. PMID- 8731930 TI - Application of electrical impedance principle in the diagnosis of diaphragm fatigue. AB - The chest and abdomen impedance respirographs (IRG), including the one dimensional IRG and the two dimensional IRG were designed and produced by applying the principle of bioelectrical impedance. Using IRG the non-synchronized chest and abdomen respiratory motions occurring in diaphragmatic fatigue were measured. The results showed that all 203 normal controls showed synchronized style of chest and abdomen respiratory motions. In 189 COPD patients 117 (61.9%) showed non-synchronized respiratory motions which could be further divided into three types: type I showed complete contra-directional respiratory movements of chest and abdomen respiration, with M > 24 % and alpha angel > 120 degrees; type II showed staggered peak of the chest and abdomen motion curves (13% < M < 24%), 50 degrees < alpha angle < 120 degrees; type III showed double peaks of abdomen trace in the one dimensional IRG and "8"-shaped double circles on the two dimensional IRG, (M < 13%, 50 degrees < alpha angle < 120 degrees. When compared with trans-diaphragmatic pressure (Pdi) and diaphragm myoelectricity frequency spectrum, the rates of conformity were 81.8% and 90%, respectively, suggesting that IRG could be reliably used for diagnosing diaphragmatic fatigue. This technique is simple, easy to use, cheap and pain-free. PMID- 8731931 TI - Correction of corrected transposition of the great arteries associated with cardiac anomalies. AB - Reported in this paper are 8 cases of corrected transposition of the great arteries associated with anomalies. All the patients underwent the repair of the ventricular septal defect and atrial septal defect, the replacement of left-side tricuspid value, the pulmonary valvotomy and the placement of extra-cardiac conduit to the pulmonary trunk respectively with no hospital death after operation. The paper also described the selection of the heart incision, the prevention of injury of the conduction system and indication of the placement of extra-cardiac conduit. PMID- 8731933 TI - Alternation of monocyte function in patients with obstructive jaundice. AB - Patients with obstructive jaundice have a high susceptibility to infection in the process of treatment and the reason for this is not fully understood. It was postulated that it may bear some relations to abnormalities of immune function. In this article, 28 cases of obstructive jaundice were selected to investigate alternation of monocyte immune function with the purpose of exploring mechanism of high susceptibility to infection from the perspective of immunology. The results showed that interleukin 1 production by monocytes significantly decreased and prostaglandin E2 increased, HLA-DR expression of monocytes was remarkably depressed. HLA-DR expression of monocytes was further decreased with recovery slower than non-jaundiced patients after operation. All this may be responsible for high susceptibility to infection in the process of treatment of obstructive jaundice. PMID- 8731932 TI - Immunoregulatory effects of indomethacin on rats with trauma. AB - In this article, we investigated changes of immune functions and immunoregulatory effects of indomethacin on rats with trauma. The results show that spontaneous suppressor T cell activity of spleen significantly increased and Interleukin I production and DNA synthesis capacity of splenocytes markedly decreased in rats with trauma. Indomethacin could markedly improve immune function, decreased spontaneous suppressor T cell activity and prompted Interleukin 2 production and DNA synthesis capacity of splenocytes. PMID- 8731934 TI - Biomechanical study on osteotomized tibias fixed with unilateral adjustable external fixator. AB - Five fresh tibias removed from 30-40 years old dead males were osteotomized at one-third supra-medium segment and then fixed by an unilateral adjustable external fixator (UAEF) to be used as a model of external fixation of tibial fracture. For each loading test under different configurations of UAEF, the ratio of the force-displacement was used to determine its stiffness value. The weakest mode of the UAEF device is in AP bending. Under each mode of compression, distraction, anteroposterial and lateral bending and torsion loading, increasing the pin number or pin diameter or pin separation, and/or reduction of pin group separation can significantly increase the stiffness values of UAEF (P < 0.01). These data revealed that UAEF is suitable for the external fixation of tibial fracture. PMID- 8731935 TI - An abduction-traction frame for the treatment of intertrochanteric fracture of femur. AB - We present a detailed outline of treatment of intertrochanteric fractures of femur with a new technique on the basis of principles of combined traditional chinese and western medicine. We use pins and an outer fixation frame, which keeps the injured limb in abduction and under traction and at the same time allows for functional exercise of all the joints in the extremity. The procedure is discussed and compared with other different techniques. PMID- 8731936 TI - The relationship of Oxalobacter formigenes and calcium oxalate calculi. AB - The intestinal Oxalobacter Formigenes were isolated in 30 cases of urolithaiasis and in 45 controls. The biologic characters and morphology of the bacteria were also observed. The results showed that the colony counts in urolith group 9 (mean 10(3)/g. faeces) were significantly less than that of controls (mean 10(8)/g. faeces) (P < 0.001). It is believed that the lesser amount of oxalobacter formigenes in urolith was the important factor of the calcium oxalate calculi formation. PMID- 8731937 TI - Forensic pathological expertise of medical tangle analysis of 139 autopsy cases. AB - In this article, we report 139 autopsy cases of medical tangle, which were examined by the Department of Forensic Pathology, Faculty of Medicine, Tongji Medical University from Jan. 1972 to Dec. 1992. The emphasis is put on analyses of the status of medical tangle autopsy in forensic pathology, involved medical departments, distribution of victim's age and sex, differences between clinical diagnosis and autoptical conclusion, the composition and causes of medical tangle. The associated problems in forensic pathological autopsy are also discussed. PMID- 8731938 TI - Oxidatively modified very low density lipoprotein enhances monocyte adhesion to endothelial cells. AB - Very los density lipoprotein (VLDL) was incubated with CuCl2(10 mumol/L) at room temperature for 24 hours. The thiobarbituric acid reactive substance (TBARS) was much higher and the electrophoretic mobility was much faster in VLDL after incubation with CuCl2 than that in VLDL without incubation with CuCl2. It demonstrated that VLDL was oxidatively modified by Cu2+. Endothelial cells were pretreated with normal VLDL (N-VLDL) and oxidatively modified VLDL (O-VLDL) and then the adhesion of monocyte to endothelial cells was assayed. We observed that O-VLDL at all the concentrations used enhanced monocyte adhesion to endothelial cells significantly. The results suggest that oxidative modification of VLDL may play a role in the early stage of atherogenesis by increasing monocyte adhesion to endothelial cells. PMID- 8731939 TI - Detection of replicative form of HCV RNA in peripheral blood leukocytes and its clinical significance. AB - Nested RT-PCR, done by using degenerated primer pair, was used to detect hepatitis C virus RNA (HCV RNA) in serum, plasma, liver and peripheral blood mononuclear cells (PBMC) of 30 patients with acute and chronic posttransfusion hepatitis C and 7 asymptomatic anti-HCV positive subjects. The results showed that the percentages of both the plus and minus strands of HCV RNA in PBMC of the patients with chronic hepatitis C was significantly higher than that with acute hepatitis C and asymptomatic anti-HCV positive subjects (P < 0.05-0.001). In 17 patients who were subjected to biopsy, the positive rate of the both strands of HCV RNA in PBMC of the patients with AH was lower than that of CAH (P < 0.05). In serum and plasma of all 37 cases, the minus strand of HCV RNA was not detected. Both plus and minus strands in liver of one patient with AH were positive, but the minus strand in PBMC negative. In 6 patients with CAH whose both strands in liver were positive, both strands in PBMC in 5 patients were also found. The present data confirmed that PBMC of the patients with hepatitis C were infected by HCV and the longer the infection time, the bigger the possibility of PBMC infection by HCV. The patients with active liver disease (CAH) had higher positive rate of minus strands of HCV RNA in PBMC. The results suggested that HCV may not only infect PBMC but also replicate in PBMC, and that the occurrence of minus strand of HCV RNA is associated with activity of liver disease. PMID- 8731940 TI - Construction of shuttle expression plasmid and stable expression of foreign gene in mycobacteria and E. coli. AB - By employing the pUC19 as a backbone, the regulatory and signal sequences which encode kanamycin resistance, and mycobacterial plasmid origin of replication (oriM) were cloned into the pUC19. The recombinant E. Coli-mycobacteria shuttle expression plasmid pBCG-8000 was constructed. The pBCG-8000 was able to replicate in both E. Coli and mycobacteria (including BCG) systems, and to confer stable kanamycin resistance upon transformants. The study should facilitate the development of BCG and other mycobacteria into multivalent vaccine vectors. PMID- 8731941 TI - Correlative study on the expression of p53 and DNA ploidy in acute nonlymphocytic leukemia. AB - We used the flow cytometric immunoassay to study the correlation between the rumor-suppressor gene product p53- and the DNA ploidy in 30 de novo cases of acute nonlymphocytic leukemia (ANLL). The results showed that 15 cases were positive expression for p53. As compared with p53 negative (p53) cases, the patients with positive p53 (p53+) had higher percentage of bone marrow blasts and lower peripheral leukocyte and platelet counts, which had no influence on the complete remission rate. Before treatment, DNA diploidy was seen in 18 cases including 12 p53- cases, and DNA aneuploidy in 12 cases including 9 p53+. After therapy, aneuploidy could be transformed into diploidy. Patients with P53+ or having aneuploidy in complete remission were at risk for early relapse. We believe that p53 may be involved in the process of leukemogenesis and progression of ANLL. PMID- 8731942 TI - Infectivity and risk factors of hepatitis C virus transmission through sexual contact. AB - HCV infection among heterosexuals with multiple partners and family members of patients with hepatitis C, and HCV RNA in the body fluid of these patients were investigated. The results showed that the HCV infection in heterosexuals with multiple partners, which was related to sexual activity, was much higher than that of healthy pregnant women. The HCV RNA in the saliva, semen or vaginal discharge of patients with hepatitis C was at detectable level. Among the patient's relatives, none of the children but 2 spouses were found to be infected with HCV. We concluded that sexual contact might play some role in HCV transmission. PMID- 8731943 TI - A new method of superoxide free radical determination in ischemia and reperfusion injury of rat liver. AB - The present study was designed to utilize the chemiluminescence (CL) method for O2- analysis in ischemia and reperfusion of the whole rat liver. The results indicated that the O2- level was lower after 30 and 60 min ischemia. After 40 min reperfusion, the level of O2- increased obviously, reaching about 8.3-fold (30 min ischemia) and 9.0-fold (60 min ischemia) (P < 0.001, P < 0.01) respectively. The results also show that Chinese Traditional Medicines ligustrazine and salvia compound can scavenge O2- effectively. PMID- 8731944 TI - The application of prostaglandin E1 indirect portal vein angiography and color Doppler ultrasound in patients with post-devascularization portal hypertension. AB - The selection of proper treatment is based on identification of the causes of massive hemorrhage of gastrointestinal tract after pericardial devascularization. The combined use of prostaglandin E1 indirect portal vein angiography, DSA of celiac artery and color Doppler can provide detailed information about portal vein system, including the presence of embolism, spongioid changes and devascularization of left gastric vein and left gastric artery and the direction of blood flow. If these techniques failed to reveal the causes of digestive tract bleeding, the endoscopy may show lesions of gastric mucosa, which could be accountable for the bleeding. PMID- 8731945 TI - The retrospective study on high risk factors of subphrenic infection after hepatectomy. AB - In order to identify the high risk factors that determine subphrenic infection after hepatectomy, 187 liver resections performed during January, 1985 to December, 1990 in Department of Surgery of Tongji Hospital were reviewed. Thirteen patients developed subphrenic infection with the incidence of subphrenic infection being 6.95%. The results of retrospective study on 187 liver resections demonstrated that the high risk factors related to subphrenic infection after hepatectomy were as follows: (1) The extent of liver resection was associated with subphrenic infection. (2) The incidence of subphrenic infection after liver resection of primary liver cancer patients with hepatic cirrhosis was higher than that of non-cirrhotic patients. (3) Intraoperative estimated blood loss greater than 1500 ml was found to be a significant risk factor in the development of postoperative subphrenic infection. (4) Adequate postoperative drainage of subdiaphragm and the raw surface of the liver was one of critical factors of decreasing subphrenic infection after liver resection. PMID- 8731946 TI - [Arrhythmogenic right ventricular disease on a young peasant]. AB - This article, together with literatures, discusses one case of arrhythmia right ventricular dysplasia (ARVD) we treated. This disease should be taken into account in some arrhythmias from right ventricle, especially refractory arrhythmias in youths. Some non-invasive techniques such as echocardiography, cardial single photon emission computed tomography (SPECT), and magnetic resonance imaging (MRI) play more and more important role in the diagnosis of ARVD. These techniques can provide specific data for the early diagnosis. PMID- 8731947 TI - Study on in vivo release of levonorgestrel-containing silicon capsule. AB - The in vivo release rate of levonorgestrel-containing silicon capsule (in vitro releasing rate: 6 micrograms/d) placed in rabbits was determined. The results showed that the average release per year was 8.32 micrograms with r being 0.96000. The release pattern of the levonorgestrel-containing silicon capsule system was proved to be zero-order release. Our study provided some experimental data for clinical use of the system for birth control. PMID- 8731948 TI - Enzymatic histochemistry of retina with experimental intraocular pressure elevation in rabbits. AB - To investigate the pathogenesis of retina lesions caused by intraocular pressure elevation, activities and distribution of enzymes in retina including lactic dehydrogenase (LDH), succinate dehydrogenase (SDH), adenosinetriphosphatase (AT Pase), acid phosphatase (ACP), cholinesterase (ChE), cytochrome oxidase (CCO), nucleotidase (5'-Nase) and glucose-6-phosphatase (G6Pase) were determined histochemically in 30 rabbits. It was found that 1) in the early stage of intraocular pressure elevation, the activities of LDH, SDH, ATPase, ACP, and ChE in retina were increased, while the activities of CCO, 5'-Nase decreased; 2) in the late stage of intraocular pressure elevation, the activities of all these enzymes but ACP, which showed a reduced activity, were close to the normal level; 3) in superoxide dismutase.(SOD-CCE) treated group, except the slight increase of LDH and G6Pase activities, the activities of the remaining enzymes were near to normal. Our results suggest that the various histochemical changes in retina induced by intraocular pressure elevation were compensatory in the early stage and were beneficial to the supply of energy needed in retinal tissue and cellular metabolism; while in the late stage, the lesion of retina cells developed due to decompensation. SOD-CCE could alleviate the retinal lesions caused by intraocular pressure elevation, and can be used as auxiliary drug for the treatment of intraocular pressure elevation. PMID- 8731949 TI - Left coronary artery fistula identified by color Doppler flow imaging. AB - Coronary artery fistula (CAF) is an abnormal communication between a coronary artery and a cardiac chamber, great vessel, or other vascular structures. The presence of the fistula is usually identified by angiography. In this paper, the diagnosis of left coronary artery-right ventricle fistula was made by color Doppler before angiography? and it was confirmed by surgery. Color Doppler flow imaging is a non-invasive method which can reveal the proximal dilatation, the course and the draining site of CAF. PMID- 8731950 TI - The combined effects of high temperature and carbon monoxide on heat stress response. AB - In this study, we have examined the effects of exposure to high temperature, carbon monoxide or a combination of both conditions in a model system, the rat and in industrial workers. In the rat liver, HSP70 mRNA and HSP70 synthesis were measured by dot hybridization and western blot. The results showed that after a heat stress HSP70 mRNA and its product, HSP70 increased significantly and there was a synergism in the combined effects of high temperature and carbon monoxide exposure on the induction of HSP70 mRNA and HSP70 synthesis. Heat played a major role in this induction. The presence of antibodies to human HSP27, HSP60, HSP70, HSC73, HSP89 alpha and beta in workers exposed to heat, carbon monoxide was also measured by western blot using purified HSPs as antigens. Plasma free amino acids were measured in the same group of workers. The incidence of antibodies to HSP27 and HSP70 was significantly higher in the workers working in an environment with extreme heat, and high carbon monoxide emission than in a control group. The carbon monoxide exposed group showed the highest incidence of antibodies to HSPs. Although our previous results indicated that workers had an insufficient protein intake, plasma free amino acids tended to increase, especially in methionine and tryptophan two kinds of amino acids which are absent from the main stress protein, HSP70. These results suggest that the major problems that these workers may face are how to facilitate the use of plasma free amino acids and reduce the inhibition of synthesis of normal proteins when they are exposed to occupational harmful factors. These results also add new information on the measurement of HSPs as a potential biomonitor to assess whether organisms are experiencing metabolic stress within their environment. PMID- 8731951 TI - Application of anticancer agent embolism in Miles' operation. AB - Presented in this paper is a report of the prospective studies on rectal cancer. By using the method of anticancer agent gelatin sponge to form embolism and by ligating the bilateral arteria iliaca internal, we performed Miles' operation on 64 cases of rectal cancer. Only 24 cases in the control group had Miles' operation. The amount of blood transfused during operation, duration of operation, days of hospitalization, infection rate of incision on the peritoneum, and relapse rate in treated group were significantly different from those in the control group (P < 0.05). PMID- 8731952 TI - Clinical application of modified Anyang type 2 esophageal intubation instrument- a new esophageal intubation method. AB - The preliminary experience in the clinical use of Anyang Type 2 esophageal intubation instrument, designed by ourselves, in 11 patients with severe esophageal obstruction due to advanced carcinoma of the esophageal was reported. The diagnosis of all 11 cases was confirmed by barium-meal roentgenography. After intubation, all the patients could take liquid or soft diet by mouth. There was no operative mortality and the patients survived 2 to 14 months (mean 5.7 months). All of them could take food by mouth before death. The clinical application proved that the intubation has been a safe and suitable management. PMID- 8731953 TI - Study of gastric and gallbladder kinetics with real-time ultrasonography in cases of functional dyspepsia. AB - Gastric and gallbladder emptying in 113 patients with functional dyspepsia (FD) were evaluated by real-time ultrasonography (RUS) after a liquid-fat meal by the patients, and compared with 15 healthy volunteers. The results showed that in FD group 69 patients (61.06%) had delayed gastric emptying, and 28 patients (24.77%) had gallbladder hypokinesia. Among them both delayed gastric emptying and gallbladder hypokinesia were found in 11 patients (9.7%), 44 patients (38.93%) had normal gastric emptying and 85 patients (75.22%) had normal gallbladder emptying. PMID- 8731954 TI - Quantitative analysis of the tomographic technetium-99m MIBI (99mTc-MIBI) myocardial bullseye display: application to diagnosis of coronary artery disease. AB - In this study, the 99mTc-MIBI myocardial bullseye display of 31 healthy persons and 34 patients with myocardial ischemia and 17 patients with myocardial infarction were analyzed quantitatively; and compared with the results of myocardial tomography analysis and qualitative bullseye analysis. The sensitivities of the three methods were 88.2%, 91.2% and 94.1% respectively (P > 0.05), and the specificities were 93.5%, 83.9% and 83.9% respectively (P < 0.05). On the other hand, the quantitative analysis obviously outperformed the other two methods in the detection of ischemic segments of myocardium near infarction zone (P < 0.01). The quantitative analysis of 99mTc-MIBI myocardial bullseye (quantitative bullseye) was an objective, specific and sensitive method for diagnosis of coronary artery disease. PMID- 8731955 TI - Analysis of plasma fibrinolysis in the patients with acute cerebral infarction. AB - In the present study, the plasma fibrinolytic activity in 30 cases of cortical artery territory cerebral infarction (CACI) and 32 cases of perforating artery territory cerebral infarction (PACI) within 3 days after ictus, and 30 sex- and age-matched controls without cardio-cerebrovascular diseases were evaluated by a comprehensive panel of assay, including the plasma tPA activity, PAI activity, endothelial capacity of tPA release and PAI/tPA ratio. The results showed that the plasma fibrinolysis and the endothelial potential to release tPA responding to stimulation in both subtypes of the patients were significantly lower than those in the controls, which provide the theoretical basis for carrying out the thrombolytic therapy in the ischemic stroke. And the study also suggested that increased plasma PAI activity could increase the risk of AS thrombotic events with increased serum triglyceride level. PMID- 8731956 TI - A comparative study of radiofrequency ablation in unipolar and bipolar fashion. AB - In this study we compared the effects of radiofrequency (RF) energy applied to the swine endocardium in a unipolar fashion and in a bipolar one with two different interelectrode distances (5 mm, 10 mm). RF energy (500 kHz) delivered to the swine endocardium was divided into eight categories: 100 J, 101-200 J, 201 300 J, 301-400 J, 401-500 J, 501-600 J, 601-1000 J, and > 1000 J. The results showed that when RF energy was applied in a bipolar fashion, the lesions involved the catheter/tissue interface and partly the interelectrode spacing, while in a unipolar fashion. They were found in the catheter/tissue interface only. At any energy level, there were no statistically significant differences in lesion depths among all the three fashions, and the lesion surface areas produced by the bipolar fashion (with 5 mm or 10 mm interelectrode spacing) were all greater than those by the unipolar fashion (P < 0.05). When the delivered energy was under 500 joules, a greater lesion surface area was found in 5 mm bipolar fashion than in 10mm bipolar fashion (P < 0.05), while energy exceeded 500 joules, the differences in the lesion surface areas were no longer significant between these two bipolar fashions. PMID- 8731957 TI - Experimental studies on preventive effects of API0134 on vessel reocclusion after thrombolysis. AB - 24 model rabbits with femoral arterial thrombosis were divided into two groups: the treatment group consisting of 12 rabbits which received API0134, and the control group composed of another 12 rabbits. 2 hours after recanalization by urokinase thrombolysis, reocclusion occurred only in 1/12 vessel (8%) with incomplete occlusion in the treated group, but in 8/12 (67%) with complete occlusion in the control group as assessed by angiograsphy. Pathological examination of specimen taken 24 hours after thrombolysis showed that 6/12 (50%) of the treated group gave the evidence of thrombus occlusion, and milder intimal injury and less adhered blood cells than in the control group, 83% of which had thrombus occlusion. In comparision with the control group, the function of platelet in the treated group demonstrated lower platelet aggregation rate (PAgR) and plasma thromboxane A2 (TXA2) level, higher prostacyclin (PGI2) and plasminogen activator (PA) activity as well as lower plasminogen activator inhibitor (PAI) activity. From the above it may be concluded that the preventive effect of API0134 on reocclusion might be due to inhibition of platelets aggregation and promotion of fibrinolysis. PMID- 8731958 TI - Localization and biodistribution of conjugate ATG-Dex-DNR in nude mice as models for human leukemia. AB - 131I-labelled anti-thymoglobuline (ATG), 131I-labelled immunoconjugate ATG-Dex DNR and 131I-labelled Ts-MoAb as control antibody, respectively, were injected by intraperitoneal (i.p.) administration into nude mice used as models for human T cell leukemia. SPECT imaging was performed from day 1 to day 8 following i.p. injection. The results showed that radioimmunoimaging of human tumor xenografts was clearest day 3 after injection in both of ATG and ATG-Dex-DNR groups, whereas it's not the case in Ts-MoAb group. Nude mice were killed 8th day after injection with antibody or conjugate. The tumor, as well as different dissected normal organs including heart, liver, lungs, kidney, femur and intestine, were harvested, weighed precisely, and radioiodine-counted. T/NT ratios in experimental group was greater than 1.0 (ranged from 1.246-7.865), and in control group they were less than 1.0 (ranged from 0.263-0.757, except for tumor/femur ratio). Our results indicated that ATG and ATG-Dex-DNR had specific affinity to cell line of T-cell leukemia CEM. PMID- 8731959 TI - The influence of intracellular cAMP on T cell proliferation. AB - The role of AC-cAMP-PKA signal system in T cell activation was investigated by lymphocyte proliferation stimulated by anti-CD3 McAb. The results indicated that AC, cAMP and PKA play important roles in determining the responses of lymphocytes to the stimulation. Activation of AC and increase of intracellular cAMP could obviously decrease lymphocyte proliferation, and lymphocyte DNA synthesis was increased to some extent by the pretreatment of PKI. PMID- 8731960 TI - The clinical application of human bone matrix gelatin. AB - This paper reports the results of 24 cases of bone defect resulting from bone tumor or tumor condition excision, and of posterior spinal fusion, treated by human bone matrix gelatin. The success rate of bone defect repair and spinal fusion is 91.67%. The results suggest that human bone matrix gelatin has excellent osteoinductive effect and is ideal substitute for bone autografts. PMID- 8731961 TI - The effects of calcium-regulating-hormone on BMC in patients with CRF. AB - By means of single photon absorptiometry, bone material content (BMC) on centerbrachium was detected in 34 patients with chronic renal failure (CRF) and 34 sex-age-matched normal controls, and their serum parathyroid hormone (PTH), calcitonin (CT) were also measured by radioimmunoassay. It was found that (1) the mean BMC was obviously lower in patients with CRF than that in the normal controls; (2) PTH and CT obviously rose in patients with CRF; (3) the decrease of BMC was strongly correlated with that of creatinine clearance (Ccr). Furthermore, the more Ccr decreases, the more PTH in blood increases; no relations have been found between BMC and the serum levels of CT. The authors suggest that the development of osteoporosis with CRF is due to the secondary increase of PTH in blood but the effect of increased serum CT levels was not so obvious. Therefore, lowering blood PTH concentrations by parathyroidectomy or renal transplantation may be beneficial to curing osteoporosis of patients with CRF. PMID- 8731962 TI - Comparison of effects of MPTCP and PTCP ceramics implanted for the treatment of bone defects in rabbits. AB - The magnetic porous tricalcium phosphate (MPTCP) and porous tricalcium phosphate (PTCP) ceramic cylinders were implanted into right and left bone defects of rabbits' radii in order to determine the utility of the MPTCP ceramics. Based on naked eye inspection, light and scanning electron microphotography, roentgenography, quantitative histological measurement of new bone formation and anti-break test for a period of 5 months. The results showed that the two kinds of ceramics were biocompatible with human tissue. MPTCP ceramics could induce more new bone formation than PTCP ceramics. Treatment of fractures with synthetic calcium phosphate ceramics and magnetic fields were discussed. PMID- 8731963 TI - The relationship between the morphology of nucleus of liver cells and the liver functions and prognosis of portal hypertension due to hepatic cirrhosis. AB - The morphology of nucleus of liver cells from 30 patients with portal hypertension due to hepatic cirrhosis and 5 normal persons were measured using an image analyzer coupled with a computer. It was found that the diameters, perimeters, areas and form factor (FF) of the nucleus of liver cirrhosis portal hypertension patients were significantly increased as compared with those of the normal subjects (P < 0.05 or P < 0.01). There was a very significant difference in this parameters between the normal persons and patients with Child-Pugh A liver function or patients with Child-Pugh C liver function (P < 0.01 for both). Significant difference in these parameters existed between the normal persons or patients with Child-Pugh A liver function and patients with liver function of Child-Pugh B (P < 0.05). No significant difference in the parameter of optic density (OD) were found between the normal persons and patients with impairment of liver function of varying degrees (Child-Pugh Classification) (P > 0.05). Our results suggest that the hepatocytes of patients with portal hypertension due to hepatic cirrhosis became juvenile and the morphology of the hepatocytes of patients with impairment of liver function of Child-Pugh C changed obviously. The enlargement and sparsity of nucleus of hepatocytes as revealed by pathological examination is a sign of severe impairment of liver function. PMID- 8731964 TI - Clinical studies on ultrasound lithotripsy in intrahepatic bile-duct via abdomen. AB - 15 patients with left/right intrahepatic bile-duct calculi received ultrasonic lithotresis via abdomen by using the percutaneous cholecystoscope and ultrasonic lithotresis apparatus. The results indicated that all the surgical manipulations were successful with a residual rate of 6.7%, compared with a residual rate of 78.2% in 23 cases of left/right intrahepatic bile-duct lithiasis treated simultaneously by lithotomy apparatus. The differences between them were statistically significant (u = 4.29, P < 0.01). Among 12 cases undergoing follow up observation, no side effect due to ultrasonic lithotresis and no residual calculus after 2-4 check-ups with B-mode ultrasound diagnostic set were found and all of their symptoms disappeared except for 1 case. PMID- 8731965 TI - The influence of thymectomy on germ-cells of the testis in mice. AB - We performed thymectomy on the immature and mature male mice and made a quantitative assay of various germ cells of the seminiferous epithelium in the testis with Image Analyser System 35 days following thymectomy. The results indicated that as compared with the control group, all germ cells of the spermatogenesis lineage decreased in the immature group and in the mature group after thymectomy. The present study showed that thymus and its hormone stimulated not only the mitotic division, but also the meiotic division of germ cells during spermatogenesis, suggesting that thymus-sexual axis may play an important role in the process of spermatogenesis in the testis. PMID- 8731966 TI - The effects of "huoxuequyu" decoction on lipid peroxidation in IUGR rats. AB - The rats exposed to passive smoking were used as the animal model to further study the pathogenesis of IUGR and the effects of "Huoxuequyu" decoction. The results showed that the mean birth-weight of fetus and SOD activity of placenta and plasma in model rats were significantly reduced, and LPO levels of placenta and plasma were significantly increased. We found that the mean birth-weight and SOD activity in treated group were higher than that in model group, and LPO levels of placenta and plasma were lower in treated group than that in model group and there was no difference in SOD activity and LPO levels of placenta and plasma as well as the mean birthweight between treated group and control group, indicating that "Huoxuequyu" decoction can protect placenta and plasma against lipid peroxidation, and improve the blood circulation of uteroplacenta, thus contributing to the increase of the mean birth-weight. PMID- 8731967 TI - Study on the effects of "foshousan" plus danshen in preventing IUGR rats with passive smoking from peroxidation in erythrocyte lipid. AB - The pregnant rats with cigarette smoking exposure were used as the animal model for studying the pathogenesis of IUGR. The mechanism of preventing intrauterine growth retardation with "Foshousan" plus Danshen by protecting erythrocytes against lipid peroxidation damage was also preliminarily discussed. The erythrocyte membrane lipid peroxide content, erythrocyte SOD activities were detected by ultraviolet split-beam spectrophotometry and chemiluminescence technique. The results showed that the erythrocyte MDA levels and ratio of abnormal erythrocyte in model group were significantly higher than those in control group and treated group, and the fetal mean birth weight, erythrocyte SOD levels were all statistically significantly lower than those in control group and treated group. The fetal birth weight showed a significant positive correlation with SOD levels and a significant negative correlation with MDA level. The results suggest "Foshousan" plus Danshen can protect erythrocytes against lipid peroxidation damage, thus conserving the normal form, structure and function of erythrocytes, improving the uteroplacental blood flow and thereby increasing the fetal mean birth weight. PMID- 8731968 TI - High performance liquid chromatography separation and simultaneous fluorometric detection of trace amount of polyamines in cell culture media. AB - A highly sensitive and precise method for the determination of the polyamines (including putrescine, spermidine and spermine) in cell culture media is described. The samples were concentrated by C18 column and the polyamines were converted to fluorometric derivatives with DNS-Cl. The polyamine derivatives were then completely separated by HPLC and determined by simultaneous fluorometric detection. The CV of intragroup and intergroup were 2.49%-4.26% and 4.29%-5.16%, respectively. The rate of recovery was 103%-99%. There was trace amount of polyamines detected by this method in the media of F12, 8900, RPMI-1640, DMEM and M199 even without incubation with cells. So this method can be used for detecting the changes of polyamines in a medium before and after incubation with cells. It is helpful for the researches on the regulation of cell proliferation by polyamines. PMID- 8731969 TI - [Expression of the extracellular domain (1-341) of rat ovarian lutropin receptor in insect cells and preliminary characterization of the expressed product]. AB - Extracellular domain residues 1-341 (designed R 341) of luteinizing hormone/human chorionic gonadotropin receptor (LH/hCG receptor) has high binding affinity for ligand. This paper describes expression of cDNA coding for R 341 in insect cells and preliminary identification of the expressed protein. SDS-PAGE silver staining and immunoblotting analysis show expressed product appears in two bands, major band has molecular weight 38.5 Kd, and weak band, 40.0 Kd. Ligand binding immunoblotting and 125I-hCG-binding blotting analysis indicate that expressed product R 341 has specific binding affinity for ligand. Ligand binding assay and Scatchard analysis indicate that recombinant receptor R 341 has high binding affinity for hCG and Kd is 5.68 x 10(-10) mol/L. PMID- 8731970 TI - [Elimination effect of ethyl alcohol on the DCCD-induced inhibition of hydrolytic activity of H+-ATPase complex]. AB - 12.5% ethyl alcohol was added into the reaction system containing mitochondrial H(+)-ATPase complex of pig heart, which was preincubated with 0.5 microgram/ml DCCD dicyclohexylcarbodiimide) at 30 degrees C. Or the DCCD and ethyl alcohol were simultaneously incubated with H(+)-ATPase at 30 degrees C. In either case, the inhibition of the hydrolytic activity of H(+)-ATPase caused by DCCD could be completely eliminated in the presence of ethyl alcohol. If methyl alcohol was instead of ethyl alcohol, the DCCD inhibition could only be partly eliminated. In the replacement of ethyl alcohol by dimethyl sulfoxide, no elimination could be observed. After preincubation of 2 micrograms/ml oligomycin with H(+)-ATPase complex instead of DCCD, the same concentration of ethyl alcohol could not caused elimination effect, which indicates no un-coupling effect happened by ethyl alcohol. The kinetic experimental result showed that ethyl alcohol exhibits non competitive inhibition to the hydrolytic activity of H(+)-ATPase complex. It was deduced that ethyl alcohol could result in conformational change of F1 of the complex, such as to affect the activity of the enzyme. The measurement of DPH (diphenylhexatriene) fluorescence polarization, the fluorescence labelled with N (1-pyrenyl) maleimide and intrinsic fluorescence of H(+)-ATPase complex compared with control show that the three cases, i.e. only treated with DCCD, only treated with ethyl alcohol or treated with DCCD and ethyl alcohol, appear different conformations of H(+)-ATPase complex. But the conformation caused by DCCD and ethyl alcohol was more like that by ethyl alcohol. This is consistent with results obtained from activity of DCCD plus ethyl alcohol and only ethyl alcohol. These results mentioned above indicate that the mechanism of ethyl alcohol eliminating the DCCD-induced inhibition of H(+)-ATPase is a conformational interaction caused by DCCD and ethyl alcohol. PMID- 8731971 TI - [The induction and differentiation of a human teratocarcinoma cell line (PA-1) in vitro]. AB - PA-1 cell line is derived from human ovary teratocarcinoma. When it grows in 10% fetal calf serum, it can be induced to differentiate by 10(-5) mol/L retinoic acid (RA). Some morphological changes can be observed after RA induction. By immunostaining of cultured cells, we found that the expression and distribution pattern of desmin and some extracellular matrix molecules, such as fibronectin, laminin and tenascin, had been changed after induction, and these changes were associated with the morphological changes. Cell growth study showed that RA treatment had no effects on growth and autocrine activities of PA-1 cells. These results suggest that some PA-1 cells were induced to differentiate along muscle cells pathway by RA. PMID- 8731972 TI - [The molecular cloning of mouse centromeric DNA (SFA-DNA)]. AB - The centromeric DNA (SFA-DNA) isolated from enriched mouse centromeric preparation has been cloned. The vector adopted for the cloning was ELBL3. About two thousand clones were obtained. The average size of the SFA-DNA inserts was estimated by measuring the inserts obtained from a mixture of 20 radomly picked clones. It was found that the average size of the SFA-DNA inserts is approximately 14 Kb. PMID- 8731973 TI - [Platelet factor 4 acts as both inhibitor and protector of hematopoietic precursor cells: possible mechanism of action]. AB - We have previously shown that platelet factor 4 (PF 4) is a potent inhibitor of megakaryocytopoiesis and that it may protect stem cells from 5-fluorouracil (5 FU) cytotoxicity. In the present work, the effects of human PF 4 on megakaryocyte (MK) growth from human CD34+ cord blood (CB) cells were studied in comparison with transforming growth factor beta 1 (TGF-beta 1). Development of MK from CD34+ cells in both plasma clot culture and liquid culture was significantly inhibited by PF 4 (5 micrograms/ml) and TGF beta 1 (1 ng/ml). Inhibition of cell growth by PF 4 was reversible judging from the fact that the CD34+ cells preincubated with PF 4 could regenerate colonies after washing and replating into the cultures. By contrast, TGF-beta 1 pretreated CD34+ cells gave rise to few colonies following replating. Moreover, incubation of CD34+ cells with PF 4 in liquid culture caused an increase in the number of both stem cell factor (SCF)-binding cells and CD34 antigen-bearing cells, and exhibited greater capacity to form MK colonies than control after the treatment of 5-FU. In vivo in mice, twice injections of PF 4 at 40 micrograms/kg with an interval of 6 h followed by one injection of 5-FU at 150 mg/kg resulted in a significant increase in the number of colony-forming cells with high proliferative potential (HPP-CFC) and colony-forming unit-megakaryocyte (CFU-MK) in bone marrow. In exponentially growing human erythroleukemia cells (HEL), the addition of PF 4 prolonged cell cycle progression and therefore resulted in an increased cell population in S phase, as determined by flow cytometric analysis. Different from PF 4, TGF-beta 1 blocked more cells in G 1 phase. These results demonstrate that PF 4 and TFG-beta 1 inhibit MK development from CD34+ CB cells by different mechanisms and suggest that PF 4, unlike TGF beta 1, exerts its inhibitory effect on cell growth in a reversible and S phasespecific manner by which it protects stem cells and MK progenitor cells from 5-FU cytotoxicity. PMID- 8731974 TI - [Making unremitting efforts and going forward]. PMID- 8731977 TI - [Progress in the study of the dorsal motor nucleus of the vagus nerve]. AB - Dorsal motor nucleus of the vagus nerve (DMV) is an important visceral motor and sensory nucleus which establishes widely connections with the central nervous system and peripheral viscera. The DMV together with the solitary tract nucleus and the area postrema form the so-called vagal sensory and motor center. There is a nerve-CSF circuit on the neurons of the DMV. The CSF-contacting neurons of the DMV could receive chemical message directly from the blood and the CSF. The DMV contains a great variety of neurotransmitters including acetylcholine, catecholamine, neuropeptide, etc. and their receptors. The DMV is involved in the central regulation of gastrointestinal, cardiovascular and endocrine functions. PMID- 8731976 TI - [Control of growth and expression of protooncogenes in regenerating liver]. AB - There are many humoral factors involved in the control of growth in regenerating liver. The complete hepatocyte mitogens such as hepatocyte growth factor (HGF), hepatic stimulator substance (HSS) can strongly stimulate hepatocyte DNA synthesis and mitosis. The hepatocyte growth inhibitors such as transforming growth factor beta 1 (TGF beta 1), however, do not stimulate DNA synthesis, but inhibit EGF mitogenesis. In addition, the comitogens such as norepinephrine and insulin are necessary to regulate the growth of regenerating liver. It has become clear that the hepatocyte proliferation and protooncogenes are linked closely. Some protooncogenes can express specifically as markers in the different phases of the cell cycle and in hepatocytes that enter the cell cycle (G0 to G1 transit) and continue to progress. PMID- 8731978 TI - [The molecular biology studies of water channels]. AB - Water channels are the aqueous pores in the plasma membranes of mammalian and plant tissue cells. The cDNA sequence of the first water channel protein (AQP CHIP) was reported in 1991. Up to date, at least five mammalian water channels are identified and referred to as the "aquaporin" (AQP). AQPs are the members of major intrinsic protein (MIP) family. Channel-mediated water transport permits the movement of water in the direction of an osmotic gradient. The distributions of AQPs are in many tissues and most of them are expressed in the renal medulla. AQP-CHIP assembles into homotetramers in the plasma membrane and each CHIP monomer appears to form a functional water pore. PMID- 8731979 TI - [Research progress in lipoprotein (a): aspects of biochemistry and molecular biology]. AB - Lipoprotein (a) (Lp(a)) is a LDL-like lipoprotein with an additional highly polymorphic glycoprotein--apolipoprotein (a) (apo(a)). Apo(a) bears great homology with plasminogen. Lp(a) is synthesized by liver and its degradation may be carried out by a nonspecific process. Apo(a) isoform size and plasma Lp(a) levels are mainly determined by apo(a) gene. Lp(a) can easily be deposited in arterial wall. It promotes growth of smooth muscle cells and inhibits fibrinolysis. These could be the mechanisms by which Lp(a) accelerates atherosclerosis and thrombosis. The physiological role of Lp(a) has not been clearly understood. PMID- 8731980 TI - [G proteins-phosphoinositide pathways in hypertension]. AB - The second messenger inositol-1,4,5-trisphosphate (InsP3) comes from two major pathways, one is initiated by a family of G protein-linked receptors and the other by receptors linked by tyrosine kinases. These separate receptors activate phospholipase C to hydrolyse phosphatidylinositol-4,5-bisphosphate to give both diacylglycerol and InsP3. The latter then mobilizes stored calcium and promotes an influx of external calcium. The alpha subunit of a newly discovered G protein (Gq) has recently been shown to stimulate the activity of PLC-beta 1. The alpha subunits of the Gq class of G proteins includes G alpha q, G alpha 11, G alpha 14, G alpha 15 and G alpha 16. The important pathologic changes in hypertension are arteriolar spasm and wall thickening. Many vasoactive substances can induce contractile response and proliferation of vascular smooth muscle cells and increase InsP3 level. However, the hypertension does not cause any persistent change in Gq. PMID- 8731981 TI - [Development of calcium fluorescent probes and their application in life sciences]. AB - It is well known that calcium is the key second intracellular messenger. Techniques using calcium fluorescent probes have been rapidly developed during the last decade, and have become one of the most important methods to measure intracellular free Ca2+ concentrations. This trend was even more accelerated after the development of a new generation of calcium fluorescent probes and the laser scanning confocal microscopy. In this article we will review the development of calcium fluorescent probes, the mechanisms and methods of measuring intracellular free Ca2+ concentrations, the properties of new calcium fluorescent probes, and their applications in life sciences. PMID- 8731982 TI - [Studies on the anticarcinogenic and immunomodulatory actions of 4-seleno carrageenan]. AB - We have systematically observed in the present study the anticarcinogenic and immunomodulatory effects of 4-Seleno-carageenan (Se-carra) as well as their possible mechanisms of actions in vivo and in vitro. It was found that se-carra had inhibitory effects on tumor growth in vivo and in vitro. The underlying mechanisms of this tumor suppressive effect may be related with the activation of macrophages, followed by the indirect priming of lymphocytes to release effector molecules such as IL-2, to express IL-2 receptors, and to selectively suppress the synthesis of macromolecules in tumor cells. The results imply that Se-carra has dual functions of tumor cell-killing effect and immunostimulating effect, therefore may serve as a novel immuno-cytotoxic anticarcinogenic drug. PMID- 8731983 TI - [Studies on the role of nitric oxide and tumor necrosis factor in immunological liver injury in mice and effects of new anti-hepatitis compounds on the liver injury]. AB - An immunological liver injury model was established by injection of micro lipopolysaccharide (LPS) into BCG (bacilli Calmette Guein)-primed mice. It was found that nitric oxide (NO) played dual effects in the liver damage induced by BCG+LPS. The NO coming from phagocytic cells showed toxic effects while those from the other cells displayed beneficial effects. Tumor necrosis factor (TNF) released by macrophages was also implicated to be a key factor in the liver damage induced by BCG + LPS. Kupffer cells were involved in BCG + LPS-induced liver injury by releasing NO and TNF. The mechanism(s) by which the two new hepatoprotectants (SY-801 and SY-640) reduced BCG + LPS-induced liver damage may be through enhancing mouse plasma NO levels and lowering NO production and TNF expression by macrophages. PMID- 8731984 TI - [Human placental lactogen]. PMID- 8731985 TI - [Progress in the study of ciliary nerotrophic factor]. PMID- 8731986 TI - [New concept of cardiovasology: vascular remodeling]. PMID- 8731987 TI - [Strychnine/glycine cytoprotective receptor]. PMID- 8731989 TI - [The bcl-2 gene family, an important regulator of apoptosis]. PMID- 8731988 TI - [Apoptosis and aging]. PMID- 8731990 TI - [Angiotensin II and the adrenal gland]. PMID- 8731991 TI - [Advance in the regulation of megakaryopoiesis]. PMID- 8731992 TI - [Membrane currents in macrophages]. PMID- 8731993 TI - [Progress in the study of lipoxin and leukotriene]. PMID- 8731994 TI - [The retrograde messengers in hippocampal long-term potentiation]. PMID- 8731995 TI - [Magnetic resonance spectroscopy in the studies of cancer cell biology and anticarcinogen]. PMID- 8731996 TI - [One-stage reconstruction of nipple-areola: the beta flap]. AB - The main disadvantage of most methods for nipple reconstruction is shrinkage of the nipple. This paper presents a one-stage procedure using a local flap with a full-thickness skin grafting that has the advantages of high patient satisfaction. Since June 1993, we have been using a technique for nipple reconstruction by raising two opposing beta-shaped random skin flaps. This technique has been performed on 8 breasts in 4 patients. PMID- 8731997 TI - [An improved technique for preserving the nipple sensation in McKissock reduction mammaplasty]. AB - An improved technique for preserving the nipple sensation in McKissock's reduction mammaplasty is described. Altogether four patients (seven breasts) have been operated on, and the primary results are highly satisfactory. The advantages of this technique are as followings: 1) The fourth lateral cutaneous nerve of the thorax, which mainly innervates the nipple, is preserved. All the patients retain the tactile sensation and the erectile reaction of the nipple after mammaplasty. 2) The operation is so modified that the suture line is smooth and has less tension. 3) The procedure ensures that both breasts are symmetrical after the operation. PMID- 8731998 TI - [An analysis of complications after implantation of saline-filled silicone prosthesis for augmentation mammaplasty in 294 cases]. AB - Complications after implantation of saline-filled silicone prosthesis for augmentation mammaplasty were analyzed in 294 patients after being followed for 3 5 years. The complications included hematoma, infection, capsular contraction, rupture of the implant, exposure of the implant, psychataxia and poor contour etc. Conditions in which these complications occurred, causes, clinical malifestations, prevention and treatment are discussed. PMID- 8731999 TI - [Experience in management of capsular contraction through areolar incision after implantation of prosthesis for augmentation mammaplasty]. AB - Since 1990, 19 cases of capsular contraction of Baker's III, IV have been treated with partial excision of the capsule and replacement of implants through areolar incision. The result was satisfactory. After six months to eighteen months follow up observation, no capsular contraction was again found. All patients were graded Baker's I. Prevention of capsular contraction was also discussed. PMID- 8732000 TI - [Mammaplasty immediately after mastectomy for breast cancer]. AB - From March 1990 to March 1994, 7 cases of one stage breast reconstruction with silicone prosthesis after mastectomy for breast cancer were performed in our hospital. All the operations were successful and no operative complication observed. They were followed up for three months to four years. The contour was good, symmetrical, soft to touch and the patients did not complain uncomfortable sensation. The authors propose that immediate reconstruction of the breast be recommended. In this paper, the indications, methods, selection of opportune time for operation and precautions are discussed. PMID- 8732001 TI - [Effect of lysostaphin on phagocyte function in burn mice]. AB - The ability of lysostaphin to prevent immunosuppression of phagocyte was assessed in burn mice. A full thickness burn covering 20% of TBSA was created on back of mouse, Lysostaphin was given intraperitoneally (400 micrograms/kg/day) for 3 days in treatment group. The results showed that lysostaphin treatment could restore the depression of phagocytosis of chick RBC by peritoneal macrophages. In the group treated by lysostaphin, peritoneal macrophage chemiluminescence expressed much greater peak value than that in both burn and normal groups. The a-value, which is a criterion of carbon clearance, was 1.63 x 10(-2) in lysostaphin treated mice, while that in burn and normal mice were 8.13 x 10(-4) and 2.76 x 10(-3), respectively. Results of the study indicated that lysostaphin was effective in improving phagocytosis by phagocytes. PMID- 8732002 TI - [The effect of dynorphin A on intracellular cAMP and cGMP contents in spleen cells of burn rats and its relation to immune function]. AB - The lymphocyte response to Con A and IL-2 synthesis was clearly suppressed in rats after thermal injury. In burn rats, the cAMP content in lymphocytes was significantly increased, but cGMP level was not changed. The serum of burn rat could inhibit Con A-induced lymphocyte proliferation and IL-2 production. The burn rat serum could also increase intracellular cAMP concentration. Dynorphin A enhanced lymphocyte proliferation and IL-2 synthesis, increased cAMP level in lymphocytes. Naloxone could inhibit the action of dynorphin A. Moreover, ynorphin A could partly reversed suppression effect of burn serum on lymphocyte proliferation and suppressed the increase of intracellular cAMP content induced by burn and rat serum. Our results suggested that dynorphin A augmented immune function of burn rats, and the action of dynorphin A was mediated by cAMP pathway. PMID- 8732003 TI - [Changes in free calcium concentration and protein kinase C activity in activated T cells of burn mice and their significance]. AB - TBSA 10%III degrees burn mice model was used. The changes in free calcium concentration ([Ca2+]i) and protein kinase C (PKC) activity in activated T cells from burn mice, and their relationship with T cell functions was studied. The results showed that [Ca2+]i and PKC activity in activated T cells were reduced after burn and these changes were closely related to reduced interleukin 2(IL-2) mRNA and IL-2 receptor alpha (IL-2R alpha) mRNA levels, decreased IL-2 production, suppressed IL-2R alpha expression, reduced T lymphocytes transformation in T cells of burn mice. Calcium cation ionophore A 23187 and PKC activator TPA could in vitro elevate respectively [Ca2+]i and PKC activity in activated T cells of burn mice. They also increased significantly IL-2 and IL-2R alpha gene expression in T cells of burn mice, but not up to the normal control. It is suggested that reduced [Ca2+]i, PKC activity in activated T cells may be one of the causes which produce suppression of T cell functions after burns. PMID- 8732004 TI - [Oxygen transport pattern in burned patient with sepsis under inotropic support]. AB - Hemodynamic and oxygen-kinetic data of 16 burn patients with sepsis were analysed to explore relationship between oxygen transport pattern and clinical outcome after dopamine and dobutamine therapy. Two patterns of oxygen transport were shown in the 16 patients. Of them, ten (62.5%) had optimal DO2 and VO2 values (model I), and six (37.5%) had lower DO2 and VO2 values than the optimal (model II). All of 6 patients with model II developed lactic acidosis, septic shock and MOF and died. Two of 10 patients in model I developed MOF, only one died. The results indicate that, in burn patient with sepsis, the decreased response of DO2 and VO2 to inotropic therapy suggests failure of tissue perfusion, oxygen extraction and utilization, and may possibly predict the outcome. PMID- 8732005 TI - [Changes in surface microcirculation of extensor digitorum longus tendon after ischemia-reperfusion of rabbit hindlimbs]. AB - The circulation was stopped by tourniquet in one rabbit hindlimb. After 2 or 5 hr of ischemia, the hindlimb was reperfused. During the first 1 hr of reperfusion, dynamic changes in surface microcirculation of extensor digitorum longus tendon were observed by using intravital microscopy. Particular attention was paid to leukocyte-endothelium adhesion and microvascular perfusion condition. It was found that 1) the number of leukocytes adherent to the venular endothelium of ischemic tissue significantly increased following reperfusion and the degree of increases appeared to be directly related to the duration of ischemic period; 2) reperfusion after 5 hr of ischemia could result in the formation of "no reflow phenomenon" in the ischemic tissues, including "primary no reflow" and "secondary capillary perfusion failure". These results suggest that 1) leukocyte-endothelium adhesion may contribute to the development of ischemia reperfusion injury; 2) in the limbs undergone 5 hr of ischemia, not all injuries that occur after ischemi reperfusion can be attributed to ischemia reperfusion injury. The injury seen some in areas may be produced by ischemia alone, and in others ischemia reperfusion-secondary ischemia injury may be responsible. PMID- 8732006 TI - [An experimental study on changes in collagen content after homologous cartilage implantation]. AB - This experiment studied the changes in cartilage collagen content during 1-20 weeks after allografting of cartilage. The results showed that the collagen content was reduced beginning from the 3rd week, and reached the lowest point in the 4th week (P < 0.05). Then it rose continuously and attained the normal level in the 8th week (P > 0.05). The study suggests that the changes in cartilage collagen content may help us to assess the cartilage structure and the elasticity after a cartilage allotransplantation. PMID- 8732007 TI - [Analysis of collagen in burned skin with SDS-polyacrylamide gel electrophoresis]. AB - Type III collagen was determined by SDS-polyacrylamide gel electrophoresis in normal rabbit skin, skin with deep II degrees burn 72h after injury, skin one month after burn, and healed burn 1.5 month after injury. Results showed that there was a reduction of type III collagen in the process of healing of burn wounds. PMID- 8732008 TI - [Mentoplasty for common congenital mandibular deformities]. AB - Since 1987, the authors have performed mentoplasty for congenital mandibular deformities in 46 patients with the assistance of X-ray pantomoscanning. Mandibular deformities include: 1) Bilateral hyperplasia of mandibular angles and massetter hypertrophy; 2) Retrognathia or micrognathia; 3) Asymmetric mandibular deformity. In this paper, following problems are discussed: 1) The advantages of an extraoral approach in mentoplasty; 2) The importance of the X-ray pantomoscanning; 3) The specific feature of asymmetric chin; 4) The improvement of the silicon implant used in mentoplasty; 5) Evaluation of the composite operative result. PMID- 8732009 TI - [Correction of unilateral cleft lip nasal deformity with modified Bardach's technique]. AB - Twenty-nine cases of unilateral cleft lip nasal deformities were corrected with modified Bardach's technique. Overall results were satisfactory. The key to a successful and stable correction of the nasal deformity is to lengthen the columella on the cleft side and to mobilize alar cartilage from its surrounding tissue, creating a symmetric shape and length. The major advantages of the technique are lengthening of the cleft columella and creation of a symmetric and well-projected nasal tip. The surgical technique is described in details. Some considerations before and during operation were discussed. PMID- 8732010 TI - [Cervico-scapulo-dorsal reverse axial flap with pedicle including the transverse cervical artery]. AB - The anatomy of the transverse cervical artery and its branches were studied on 52 arteries in 26 cadavers. It was confirmed that blood in the transverse cervical artery might flow reversely into the suprascapular artery and the posterior intercostal perforators through anastomic branches. An axial skin flap in the cervico-scapulo-dorsal area could be adequately supplied by this artery. We have used this flap in reconstruction for burn scar contractures of the neck in 20 cases, and in one patient bilateral flaps were used. There was partial necrosis of the tip in 4 flaps, while all others survived completely. PMID- 8732011 TI - [Experience in repair of eyelid ectropion with temporofrontal flap]. AB - We used the temporofrontal flap to repair eyelid ectropion in 12 patients. The results were satisfactory. The indications, advantages and disadvantages, and the points for attention of this operation method were discussed. Experiences and lessons were summarized in this article. PMID- 8732012 TI - [Distribution of fibronectin in hypertrophic scar and its significance]. PMID- 8732013 TI - [Effect of Tripterygium wolfordii on inhibition of rejection reaction of allogeneic skin graft in mice]. PMID- 8732014 TI - [Anatomical and histological observation on soft tissue of nasal dorsum and the nasalis-procerus aponeurosis]. AB - 26 nasal specimens of cadavers were studied with anatomical and histological methods. Two layers of collagen fibre arranged in different pattern were found beneath the nasalis transversus and procerus. The superficial layer of compact collagen fibres directly attached to and wraped up nasalis transversus and procerus, and it was named the nasalis-procerus aponeurosis; the deep layer of relatively flexible collagen fibres was loose subaponeuratic tissue. The study showed that soft tissue of nasal dorsum could be divided into four layers; skin, superficial fascia, skeletal muscle and aponeurosis, and loose subaponeurotic tissue. This result provides scientific basis for proper design of rhinoplasty. PMID- 8732015 TI - [Advances in the experimental studies of hydroxylapatite-based composites]. PMID- 8732016 TI - [The development of skin external expander and clinical applications]. AB - The skin external expander that was designed and developed by the authors is a new device to produce extra skin tissue for the repair of defects by continuous traction over the skin. Clinical application in 38 cases since June 1993 have shown that the method has following advantages: easy in manipulation, unnecessary to bury the expander under the skin, safe and less complications with reliable effects. Accurate measurement of the expanded skin can be obtained with the method. Six months' follow-up of 18 cases demonstrated satisfying results. PMID- 8732017 TI - [Application of computer for measuring the skin area during external expansion]. AB - The skin external expander has been used to produce extra skin and soft tissue by continuous traction of the skin. The computer image measurement technique was designed to accurately measure the extra skin. The paper describes the technique and its clinical applications. PMID- 8732018 TI - [Full-load expansion techniques and preliminary clinical application]. AB - For accelerating the inflation process of soft tissue expansion, a continuous inflation protocol regarding pain as the criterion of over-inflation, the full load expansion technique, was introduced. It is carried out by restricting persistently the pressure in the expander near the pain threshold in a state of dynamic equilibrium between the infusing rate and the skin expansion velocity, with a simple and practical assembled device. This inflation is neither at a uniform infusing rate nor at a constant filing pressure. 19 expanders in 13 patients were inflated following this protocol. Their inflations were respectively fulfilled in 5 to 20 days, depending individually on their anatomic regions, total volumes of infusion, areas of the overlying skin and the patient susceptibilities. PMID- 8732019 TI - [Clinical application of immediate intraoperative expansion technique]. AB - Since June 1992, 14 patients with skin or soft tissue defects from resection of scar or nevus were treated using the technique of immediate intraoperative expansion with good results. The operative procedure, indications and key points are discussed. We concluded that immediate intraoperative expansion may be accepted as a simple, safe and effective technique for the repair of the skin defects, which can be sutured directly under great tension. PMID- 8732020 TI - [Clinical application of expanded thin skin flap]. AB - Fifteen patients were treated with the expanded thin skin flaps with the large area defects of soft tissue resulting from scar, pigmented nevus, hemangioma, chronic ulcer and neurofibroma. The operative procedure includes tissue expansion and defatted flap transplantation. All flaps that preserved only a thickness of 3 mm adipose tissue or superficial fascia beneath the subdermal vascular network survived with the primary healing of 12 cases and the delay healing of 3 cases in the maximal ratio of length to width 4.3:1. The details of clinical experience and primary experiment were presented. PMID- 8732021 TI - [An assessment of factors leading to the failure of clinical application of venous flap]. PMID- 8732022 TI - [Clinical study of the venous flaps]. AB - Seven venous flaps were used clinically. Of them six flaps survived completely, one saphenous venous flap had partial necrosis of the derma. The size of the largest survived flap was 5 cm x 15 cm. Three principles regarding the flap application were emphasized: 1) The tissue surrounding the recipient site should be normal. High venous pressure and oxygen tension in the recipient veins are needed. 2) Rich venous network and an axial vein are required in the recipient site. 3) Reliable fixation of the flap is necessary to avoid any damage to new blood vessels. PMID- 8732023 TI - [Application of the local flap pedicled with a superficial vein in the repair of limb wounds]. AB - Fourteen cases of limb wounds were repaired with the local flap pedicled with a superficial vein. All the flaps survived with satisfactory results. The superficial vein that speeds drainage from the flap can also provide blood supply to the flap, which is beneficial to the survival of the flap. The ratio of length to width of the flap is greater than that of the traditional flap. The creation of the flap is easy and the nearby artery is intact. The technique details are described in the article. PMID- 8732024 TI - [An analysis of burn cases in the last decade]. AB - The clinical data of 3,911 burn patients admitted to our institute from January, 1983 to December, 1992 were analyzed mainly on the distribution of age, causes of burns, burn area, combined injuries, common complications, incidence of burn shock, mortality and cause of death. Results showed that 91.7 per cent of deceased had suffered from inhalation injury, internal organ complications and systemic infection, suggesting that inhalation injury, visceral complication and systemic infection still remain to be the three major causes of death in burn patients. PMID- 8732025 TI - [Early enteral feeding and supplement of glutamine prevent occurrence of stress ulcer following severe thermal injury]. AB - Gastroscopic and gastric juice analysis studies were carried out on 12 patients with total body surface area over 30 percent who received early enteral feeding, supplemented with glutamine. The results indicated that early feeding was safe in major burn patients. All patients were found to have ASML on postburn days 1 to 2. Stress ulcer was not found in patients after several excision operations, and about 86 percent of ASML disappeared within about 2 weeks. CONCLUSION: early feeding and supplement of glutamine can prevent the complication of stress ulcer following severe thermal injury. PMID- 8732026 TI - [The characteristics of inhalation injury and pulmonary infection in burn patients and the influence on their mortality]. AB - 940 patients with thermal injury during the past 14 years were reviewed. Of them, inhalation injury was diagnosed in 75 patients, including 15 mild injury who all survived, 25 moderate injury, of whom 13 died with a mortality rate of 52.0% and 35 severe injury, of whom 31 died with a mortality rate of 88.6%. The data showed that 70% of burn that caused inhalation injury occurred in a closed space and 96% of which also caused facial injury. With the burn extent increasing, the incidence of inhalation injury and pulmonary infection rise correspondingly. The time and incidence of pulmonary infection in the patients with inhalation injury was earlier and higher than that in those without inhalation injury (P < 0.01). With similar burn area and age, the risk of death in the patients with inhalation injury was 17.2 times of the patients without inhalation injury (P < 0.001). Pulmonary infection further increased mortality of the patients (P < 0.001). PMID- 8732027 TI - [The role of complement in inhibition of intracellular bactericidal activity to P. aeruginosa of PMN in seriously burnt patients]. AB - 129 PMN-samples from the peripheral blood of 70 burnt patients were collected, and ICBA, SG and O2- were dynamically studied. The harmful effects on patients' plasma on normal human PMN, and specific blocking effect of anti-human C3. C5 serum (AHC3C5S) on the above harmful effects were also observed. THE RESULTS: 1) All the parameters values showed a significant decrease in seriously or moderately burnt patients as compared with normal values; the decrease was most marked on 1st-6th postburn days when deep burn surface exceeded 45%. 2) ICBA was significantly correlated with SG and O2-. 3) Patients' plasma greatly reduced the reserves of ICBA, SG and O2- in normal PMN, while AHC3C5S might lessen the reduction in term of net reserve rates: ICBA (67.33) > SG (51.60) > O2- (46.68). The findings suggest: ICBA levels are reversely related with DBSA and fragments of C3 and C5 are the main factors in reduction of ICBA. PMID- 8732028 TI - [Confirmation of the persistence of cultured allogeneic keratinocytes by DNA fingerprints]. AB - We have successfully used DNA fingerprint to confirm the existence of cultured allogeneic keratinocytes after transplantation. Four biopsies from two burn patients who had been grafted with cultured allogeneic keratinocytes were examined by this technique. The results showed that the cultured allogeneic keratinocytes could survive on the grafted wounds for at least 28 days post grafting. PMID- 8732029 TI - [Effect of lipid peroxide on lymphocytes and its role in postburn immunosuppression]. AB - PURPOSE: Mechanisms of postburn immunosuppression are complicated and remain unclear. In the present experiment the effect of linoleic acid hydroperoxide (LOOH) on lymphocytes was evaluated in vitro, and then the changes in postburn lipid peroxide (LPO) levels and lymphocyte functions were measured, in order to investigate the mechanism of immunosuppression following burn. METHODS: Proliferation and IL-2 production and LPO of lymphocytes were assayed after the incubation with LOOH for in vitro study. On day 6 after induction of 11%-12% TBSA full-thickness burn, the animals were sacrificed, and proliferation and IL-2 production of splenic lymphocytes, LPO levels of the plasma, livers and spleens were measured. RESULTS: LOOH inhibited proliferation and IL-2 production and induced lipid peroxidation of lymphocytes in vitro, and Vit E could attenuate the effects of LOOH. LPO levels increased and proliferation and IL-2 production decreased after burn, but those changes in Vit E or SOD group showed no statistical significance compared with normal control group. CONCLUSION: Enhancement of lipid peroxidation and increased LPO after burn may be one of the mechanisms of postburn immunosuppression. PMID- 8732030 TI - [Systemic and local administration of interferon-gamma impairs wound healing]. AB - Interferon-gamma (IFN-gamma), which is a member of cytokine family, has many physiological functions. In this study, we evaluated the effects of IFN-gamma on wound healing in a model of incised wound of rat skin. Twenty Wistar rats were divided into three groups, consisting of IFN-gamma local administration (n = 8), IFN-gamma systemic administration (n = 8) and normal saline control (n = 4). IFN gamma was administered in doses of 2,000 u/rat, and then daily. Wound areas were measured everyday. At day 10, tissue from wounds was obtained, and it was evaluated for protein amount in granulation tissue and also subjected to morphometric analysis with hemotoxylin and eosin and collagen fibril stain. We found that IFN-gamma impaired wound healing after either systemic or local administration as compared with control, as evidenced by reduction of new collagen fibril deposition and delay of reepidermidalization. On days 7 and 10 after wounding, wound area in normal saline treated rats was much smaller than those in rats treated with IFN-gamma. These results indicated that the impairing effects of IFN-gamma on wound healing must be considered when it is used in clinic. PMID- 8732031 TI - [Experimental studies on the pharmacokinetics of cefoperazone (CPZ) injection under burn eschar]. AB - In this study, the rabbits were categorized into three groups. The scald (III degrees TBS 10%-15%) was caused by exposing these area to hot water at 90 degrees C for 40 seconds. Every rabbit was injected CPZ (25 mg/kg) three times. The changes in the concentration of CPZ were measured by employing MBPD and HPLC methods and experimental data were analyzed by using pharmacokinetic computer program. RESULTS: 1) The T1/2e of CPZ injection in subeschar region was longer and the Cmax was higher than that of intravenous route. 2) The concentration in blood was shown two times higher than after subeschar injection. 3) The levels of concentration were higher than MIC'S in an area of 6.5 cm diameter 24-36 hours after injection. CONCLUSION: CPZ injection in subeschar region was the best route of treatment and prevention of subeschar infection. PMID- 8732032 TI - [The lateral thoracic free flap in facio-cervical reconstruction]. AB - We have applied the lateral thoracic free flap in 16 patients with face and/or neck defects during the past three years. 13 flaps survived completely and 3 flaps failed which was substituted with skin grafting. The lateral thoracic free flap possesses many supplying vessels and its advantages include big size, hairless skin with well matched color and texture and inconspicuous donor site that can be closed directly. The technique of hemodilution and microvascular anastomotic apparatus are introduced. The indications of the flap and the limitation of its size are discussed. PMID- 8732033 TI - [Clinical application of thoraco-umbilical skin flap]. AB - Since 1987 we have successfully used thoraco-umbilical skin flaps as free transplants to repair 8 cases of severe burn scar deformities of jaw-neck and hand regions, thoraco-umbilical insular skin flaps to each case of congenital vaginal atresia, severe vulval scar due to sulfuric acid burn and large penal tissue defect accompanied with severe urethral leakage. This flap has the following advantages: namely definite anatomical position, larger vascular diameter, fight large skin flap area, and similar skin completion as that of the jaw-neck region. Therefore they are the ideal substitute for the repair of soft tissue defects of jaw-neck region as well as extremities. The flap has long vascular pedicle, abundant vascularization, easy transportation and it is suitable to be a retrograde insular flap for repairing of vulval tissue defect, reformed penal vaginoplasty. PMID- 8732034 TI - [Clinical application of the bilobed flap with first volardorsal artery in repair of the hand]. AB - We report the successful use of the bilobed flap pedicled with the first volardorsal artery of the hand to repair the defects of the first web, thenar, vola or reconstruct the thumb in ten cases. The flap ranged from 3 cm x 10 cm to 5 cm x 13 cm. The width of the pedicle was 1.5-2.5 cm. The size of each lobe was 2 cm x 3.5 cm to 3 cm x 6 cm. The article discusses the anatomy of the flap, operation method and indications. PMID- 8732035 TI - Analysis of heterogeneity of gene products (interferon) expressed in yeast. AB - FPLC, SDS-PAGE and Western blot techniques are used to analyse the heterogeneity of interferon alpha A (IFN-alpha A) expressed in yeast. The heterogeneity consists of (i) the presence of IFN polymer, (ii) partial processing of signal leader peptide and (iii) internal degradation. The reasons for heterogeneity of gene products in expression system of yeast are analysed. The methods of avoiding heterogeneity, such as depolymerization, adding inhibitors of protease to the culture supernatant, the oligonucleotide-directed deletion mutagenesis and improvements of fermentation, are discussed. PMID- 8732036 TI - Multiobjective optimization of neural network. AB - A theoretical model of multiobjective optimization neural network (MONN) and a new learning algorithm are presented. Experiment results show their effectiveness and advantage. PMID- 8732037 TI - Differences of desensitization and hypersensitization between alpha 1A- and alpha 1B-adrenoceptors in rat isolated blood vessels. AB - AIM: To study the differences of the agonist-induced desensitization and the reserpinization-induced hypersensitization between alpha 1A- and alpha 1B adrenoceptors (AR) mediated vasoconstriction. METHODS: The thoracic aortae, mesenteric, and renal arteries of rats were isolated. The cumulative concentration response-curve (CCRC) of vasoconstriction for NE was recorded. NE activated only alpha 1-AR since the perfusing Krebs solution contained propranolol 1 mumol . L-1 and yohimbine 0.1 mumol . L-1 to block beta- and alpha 2-AR. CCRC for NE was made, preparations were pretreated with CEC 50 mumol . L-1 for then washed, and CCRC for NE was repeated. After i.p. reserpine 4 mg . g-1 i.p., the rats were killed, the thoracic aortae and renal arteries were taken, CCRC for NE was compared with the corresponding blood vessels in control rats. RESULTS: Pretreatment with CEC caused reductions of the NE-induced maximal constriction by 82.5 +/- 3.0% (P < 0.01) and 54.2 +/- 9.5% (P < 0.01) in thoracic aortae and mesenteric arteries, respectively, but no effect in renal arteries. Preincubation with NE caused the alpha 1-AR mediated-vasoconstriction diminished 14.4 +/- 5.9, 1.8 +/- 0.8 and 7.3 +/- 1.8 times in aortae, renal arteries, and mesenteric arteries, respectively. In reserpinized rats, the contraction in renal arteries induced by NE increased by 56%, but showed no change in aortae. CONCLUSION: Alpha 1B-AR mediated vasoconstriction is easier to be desensitized, while alpha 1A-AR mediated vasoconstriction is easier to be hypersensitized in rats. PMID- 8732038 TI - Effects of ONO-1078, a leukotriene antagonist, on cardiovascular responses induced by vagal stimulation, capsaicin, and substance P in guinea pigs. AB - AIM: To determine the role of ONO-1078, 4-oxo-8 -[p-(4-phenylbutyloxy) benzoylamino]- 2-(tetrazol-5-yl) -4H-1-benzopyran hemihydrate, in cardiovascular responses induced by vagal stimulation, capsaicin, and substance P. METHODS: Evans blue extravasation in the atrium and ventricle, and mean arterial pressure (MAP) were observed. RESULTS: Electric stimulation of vagus (ESV, 10 Hz, 5 ms, 2 or 10 V, for 90 s) increased Evans blue extravasation in the hearts of atropine (1 mg.kg-1, i.v.)-pretreated guinea pigs. Capsaicin (0.05 mg.kg-1, i.v.) and substance P (1 microgram.kg-1, i.v.) enhanced the dye extravasation and elicited a drop in MAP. ONO-1078 (0.03 and 0.1 mg.kg-1, i.v.) inhibited ESV-induced response, especially at stimulation of 2 V. ONO-1078 (0.03 mg.kg-1) attenuated capsaicin-induced cardiac microvascular leakage and hypotensive response, but failed to inhibit substance P-induced responses. CONCLUSION: ONO-1078 can modulate the cardiovascular responses in neurogenic inflammation, possibly mediated by inhibiting sensory neuropeptide release. PMID- 8732039 TI - Nicardipine enhances cytoplasmic calcium concentration in mouse thymocytes in vitro. AB - AIM: To study the effect of nicardipine (Nic) on the cytoplasmic free calcium concentration ([Ca2+]i) in thymocytes and on the proliferation of thymus lymphocytes from mice in vitro. METHODS: The [Ca2+]i was measured by loading cells with the fluorescent dye Fura-2. The lymphocyte proliferation was determined by [3H] thymidine incorporation. RESULTS: Nic 1-30 mumol . L-1 elevated [Ca2+]i of resting thymocytes with the percentages of its increase 43% 192% and 12%-148% in the presence and absence of extracellular Ca2+ respectively. Mitogen Con A 5 mg . L-1 also released Ca2+ from intracellular stores. Nic decreased the enhancement of [Ca2+]i induced by Con A. The IC50 (mumol . L-1) was 26.8 (95% limit: 24.8-29.0) and approximated to 10 in the presence and absence of extracellular Ca2+ respectively. Nic 1-30 mumol . L-1 did not stimulate the proliferation of resting lymphocytes from mouse thymuses (P > 0.05), but inhibited the proliferation when the lymphocytes had been stimulated by Con A (P < 0.01). CONCLUSION: Nic elevated [Ca2+]i, which decreased the response of lymphocytes to mitogen. PMID- 8732040 TI - Effects of 3-morpholinosydnonimine-N-ethylcarbamide on hypoxia-induced mechanical and electric responses of isolated pig coronary artery. AB - AIM: To study effects of 3-morpholinosydnonimine-N -ethylcarbamide (SIN-1) on hypoxia-induced mechanical and electric activities of the isolated pig coronary artery. METHODS: Mechanical tension and membrane potential were measured simultaneously. RESULTS: Hypoxia initially caused a transient vascular smooth muscle cell membrane hyperpolarization followed by a membrane depolarization in isolated pig coronary artery. Subsequent addition of SIN-1 100 mumol . L-1 or verapamil (Ver) 10 mumol . L-1 led to membrane repolarization and relaxation of the vascular smooth muscle. Nitro-L-arginine (NLA) 0.2 mmol . L-1 and KCI 40 mmol . L-1 also induced membrane depolarization and vasoconstriction, which were similarly suppressed by SIN-1 or Ver. CONCLUSION: Hypoxic contractile response in isolated pig coronary artery is mediated by an increased Ca2+ influx via suppression of nitric oxide release. PMID- 8732041 TI - Augmentation of dopamine release by (-)-stepholidine from rabbit and rat caudate slices. AB - AIM: To study the effect of (-)-stepholidine (SPD) on dopamine (DA) release evoked by electric stimulation on slices of rabbit caudate nucleus. METHODS: Slices of rabbit caudate nucleus were preincubated with [3H]DA and then perfused and stimulated electrically. RESULTS: Quinpirole (Qui), a selective DA D2 receptor agonist, reduced [3H]DA overflow elicited by 24 mA electric stimulation (IC50 = 0.12, 95% confidence limits 0.09-0.17 mumol . L-1). SPD markedly increased the potential of the stimulation-evoked [3H]DA overflow in a dose dependent manner and reversed Qui (1 mumol . L-1)-induced attenuation of [3H]DA release. The pA2 value calculated from the data of [3H]DA overflow for SPD was 7.495. In the experiments with rat caudate nucleus slices, SPD (0.1 mmol . L-1) increased the [3H]DA outflow from 3.3 +/- 0.2% to 6.5 +/- 0.5% of the tissue [3H]DA content, which was further enhanced by the protein kinase C (PKC) activator phorbol 12,13-dibutyrate (PDB, 1 mumol . L-1) to 10.1 +/- 1.0% of the total [3H]DA in the tissue. CONCLUSION: SPD is a presynaptic D2 autoreceptor antagonist and induces a synergic effect on [3H]DA release process with PKC. PMID- 8732042 TI - Effects of isoprenaline on delayed rectifier potassium current in isolated guinea pig ventricular myocytes. AB - AIM: To study the effects of isoprenaline (Iso) on the delayed rectifier potassium current (Ik) in isolated guinea pig ventricular myocytes. METHODS: Single cells were isolated from guinea pig ventricle. Ik was studied under voltage clamp conditions. RESULTS: When Ik was activated by depolarizing pulses to +40 mV of increasing duration (40-300 ms), Iso 1 mumol . L-1 caused an enhancement in Ik which was larger for longer pulses (150-300 ms). This was also seen when the intracellular calcium was buffered by 1,2-bis(2-aminophenoxy) ethane-N, N, N1, N1-tetraacetic acid (BAPTA). Intracellular application of BAPTA caused a decrease in Ik activated by longer pulses. CONCLUSION: There were two components of Ik, one of which was modulated by Iso and intracellular Ca2+. PMID- 8732043 TI - Protective effect of preconditioning on ischemic heart and characterization of adenosine receptors in ischemic rabbit hearts. AB - AIM: To assess the role of adenosine receptors in the cardioprotective effect of preconditioning and the characterization of adenosine receptors in heart. METHODS: In 18 anesthetized rabbits, myocardial ischemia was induced by occlusion of left descending coronary artery. ST segment elevation in epicardial electrogram was used as a criterion of acute ischemic injury. Myocardial adenosine receptors were measured using [3H] adenosine ligand. RESULTS: After 60 min ischemia, epicardial ST segment elevation was higher in ischemia group (13.9 +/- 0.6 mV) than in the ischemic preconditioning group (3.1 +/- 0.5 mV). In membranes of the ventricular myocytes prepared from ischemic-preconditioning group, the density of adenosine receptors was higher than that of ischemia and sham ischemia groups (Bmax being 602 +/- 40, 348 +/- 28, and 335 +/- 30 pmol/g protein, respectively). Affinity of the receptors for the [3H] adenosine was not different in the 3 groups (Kd being 181 +/- 18, 169 +/- 21, and 196 +/- 24 nmol . L-1, respectively). CONCLUSION: Activation of adenosine receptors along with an increase in adenosine receptor density during ischemic-preconditioning provides the basis for adenosine to exert its protective effect on the ischemic heart. PMID- 8732044 TI - Protein kinase C inhibitor H-7 blocks effects of tumor necrosis factor on bone cells. AB - AIM: To study the effects of tumor necrosis factor (TNF) on the neonatal mouse osteoblast-enriched calvarial cells and effects of protein kinase C (PK C) inhibitor, 1-(5-isoquinolinesulfonyl)-2 -methylpiperazine (H-7) on the TNF actions. METHODS: [3H]TdR uptake by the osteoblasts was used to measure cell proliferation. Cellular alkaline phosphatase (AIP) and tartrate resistant acid phosphatase (trAcP) activities were determined spectrophotometrically. RESULTS: TNF (1-100 kU . L-1) inhibited both proliferation and expression of AIP activity, but stimulated trAcP activity. These TNF-induced actions were blocked by simultaneous addition of H-7 (5-20 mumol . L-1). CONCLUSION: TNF has potent effects on the osteoblasts, and the blockade of TNF actions by H-7 suggests that TNF exert its effects through PK C. PMID- 8732045 TI - Effect of 5-HT on pain modulation of substance P in spinal cord of rats. AB - AIM: To study the effect of serotonin (5-HT) on pain modulation of substance P (SP) in spinal cord of rats. METHODS: Using immunohisto-chemistry and measurement of pain threshold. RESULTS: The c-fos expression evoked by intrathecal injection (it) SP 10 micrograms and sc 5% formaldehyde (For) 150 microL in the hindpaw was densely distributed in the laminae I, II, V, and VI of spinal dorsal horn. The pain threshold in the SP group was decreased while the pain intensity rating measured by behavioral method in the For group was increased. The c-fos expression induced by it 5-HT 20 micrograms was mostly distributed in the spinal dorsal horn in laminae III-IV and the pain threshold was increased. SP and For induced c-fos expressions in the spinal cord and the pain responses were reduced by 5-HT and increased by 5-HT depletor fenclonine 300 mg.kg-1. CONCLUSION: SP mainly played an algogenesia in the spinal cord. 5-HT inhibited the c-fos expression in the spinal cord evoked by SP and participated in pain modulation of SP. PMID- 8732046 TI - Antiulcer effect of diltiazem in rats. AB - AIM: To study the effects of diltiazem (Dil) on several experimental gastric ulcers in rats. METHODS: The gastric mucosa damage was induced by indometacin (Ind), restraint, pyloric ligation, and absolute ethanol in rats. Dil 5, 25, 50 mg . kg-1 ig bid for 5 times. The number of gastric ulcers, the secretion of gastric juice, hydrochloric acid (HCI), and pepsin A were detected. The production of malondialdehyde (Mal) and the activity of superoxide dismutase (SOD) in gastric mucose were examined. RESULTS: Dil 5, 25, 50 mg . kg-1 ig protected the gastric mucosa against the damages in a dose-dependent manner. Dil inhibited the secretion of gastric juice, HCI, pepsin A, and Mal production of the gastric mucosa, but increased the activity of SOD in the gastric mucosa. The production of Mal was decreased from 9.3 +/- 3.7 to 6.5 +/- 1.9 mumol/g wet weight (P < 0.05) and the activity of SOD was increased from 6.1 +/- 5.6 to 12.8 +/- 2.8 kU/g protein (P < 0.01) by Dil 50 mg . kg-1. CONCLUSION: The inhibition of gastric secretion and lipid peroxidation induced by oxygen free radicals of gastric mucosa was related to the antiulcer effect of Dil in rats. PMID- 8732047 TI - [Nicotinic currents of cultured rat superior cervical ganglion neurons and use dependent block by mecamylamine]. AB - AIM: Comparison of action of nicotinic agonists and antagonists on nicotinic acetylcholine receptor (nAChR) in superior cervical ganglion (SCG) neurons. METHODS: Whole-cell recordings were made from cultured neonatal rat SCG neurons. Cholinergic drugs were applied by local pressure perfusion. RESULTS: The neurons were activated by nicotinic agonists and peak current were acetylcholine (ACh), 443 +/- 183 pA; nicotine, 1175 +/- 377 pA; dimethylphenylpiperazinium (DMPP), 2946 +/- 358 pA, respectively. The nicotinic responses were blocked by mecamylamine (Mec), hexamethonium and curare, the efficacies were 435 +/- 154 pA, 725 +/- 320 pA, 887 +/- 214 pA, but not by alpha-bungarotoxin. The block by Mec was use-dependent, i.e., it was dependent on repeated presentation of the agonists. The first 6 peak currents were expressed as percentage of the first response as following: 100, 64 +/- 3, 50 +/- 3, 41 +/- 4, 32 +/- 3%. CONCLUSION: The present data suggest that nAChR of SCG neurons have different pharmacological characteristics from that of muscle or central neurons. PMID- 8732048 TI - [Inhibition of sodium artesunate on rat erythrocyte membrane Na(+)-K(+) exchanging ATPase in vitro]. AB - AIM: To study the effects of sodium artesunate (SA) on Na(+)-K(+)-exchanging ATPase activity of rat erythrocyte membrane in vitro. METHODS: SA of different concentrations (0, 0.5, 1, 5 and 10 mumol . L-1) were added respectively in the reaction system. The Na(+)-K(+)-exchanging ATPase activity was calculated by measuring the amount of inorganic phosphate released in the reaction system. RESULTS: Along with the increase of SA concentration (0, 0.5, 1, 5 and 10 mumol . L-1), the Na(+)-K(+) -exchanging ATPase activity decreased, the inhibitory rates were 15%, 29%, 46%, and 75%, respectively. Increasing the concentration of substrate ATP to 125, 250, 375 and 500 mumol . L-1 and determining the kinetics of enzyme. Making the Eadie-Hofstee kinetic curve by linear-regression analysis. The result revealed that the inhibitory effect of SA on the enzyme was competitive. CONCLUSION: SA affected the ion transfer of host erythrocyte membrane and membrane function. PMID- 8732049 TI - [Antagonistic effect of oleanolic acid on anaphylactic shock]. AB - AIM: To study the antagonistic effect of oleanolic acid (OA) on anaphylactic shock in guinea-pigs sensitized with 6-aminopenicillanic acid (APA)-protein. METHODS: The anaphylactic shock was achieved by i.p. of APA-protein in guinea pigs. The serum antibody titers of guinea pigs were measured with the passive cutaneous anaphylaxis test, and the pulmonary histamine by the method of fluorometric determination. The degranulation of mast cells was examined by rat calvarian periosteum method. RESULTS: In guinea pigs sensitized with APA-protein, OA 25, 50, 100 mg . kg-1 reduced anaphylactic shock rates from 100% (sensitized control group) to 70%, 18%, and 15% (OA pretreated group), and death rates from 100% to 70%, 18%, and 0%, respectively. OA had an inhibitory effect on the antibody formation and reduced the histamine contents in the lung tissues in the sensitized guinea pigs. In addition, the homologous passive cutaneous anaphylaxis in guinea pigs and the degranulation of mast cells of calvarial periosteum in rats were prevented by OA 25, 50, and 100 mg . kg-1. CONCLUSION: The antagonistic effect of OA on anaphylactic shock was related to the inhibition of antibody formation and the reduction of histamine contents. PMID- 8732050 TI - [Mutagenic effects of bimolane]. AB - AIM: To study the genotoxicity of bimolane. METHODS: Bimolane 5, 10, and 15 mg . kg-1 was injected i.p. in mice to investigate its effects on chromosome/chromatid aberrations of bone marrow cells. Mutagenic effects on TA97, TA98, TA100, TA102 were studied in Ames test. RESULTS: Bimolane 5, 10, and 15 mg . kg-1 induced of chromosome/chromatid aberrations, and the frequency of aberration cells (ACF) induced by bimolane increased markedly (P < 0.01); Bimolane without S9 showed mutagenic to TA98 and TA102 at the concentrations of 100 and 150 micrograms/plate in Ames test. CONCLUSION: Bimolane is a kind of genotoxic compound. PMID- 8732051 TI - [Construction of genomic library of L. interrogans serovar lai and preliminary study of recombinant plasmid pDC38]. AB - A genomic library, consisting of approximate 12 000 recombinants, has been constructed for the Leptospira interrogans serovar lai in E. coli. using pUC18 as the vector. Hybridization analysis with the DNA fragment containing OmpL1 gene was performed and 10 positive clones were screened from the genomic library. One of the positive clones, designated pDC38, showed hybridization signal with the DNA of 7 serovars 8 strains of pathogenic leptospires, but not with the DNA of nonpathogenic leptospires (L. biflexa), Leptonama illini, E. coli., and 2 strains of L. interrongans. PMID- 8732052 TI - [Analysis of beta-thalassemia mutations and prenated diagnosis in Chengdu population]. AB - Ninety-five of non-differential diagnostic patients were detected by dot-blot analysis on enzymatically amplified DNA with a number of allele specific oligonucleotide probes complementary to the most common mutations in Chengdu population. Prenatal diagnosis was accomplished by the same procedure on enzymatically amplified amniocyte DNA. The result revealed fifty-eight cases of beta-thalassemia. Of the 73 chromosomes tested, twenty-eight (38.4%) had the codon 17(A-->T) mutation, twenty-one (28.8%) had the codon 41-42(-TTCT) mutation, fourteen (19.0%) had the IVS-I-654(C-->T) mutation; nt--28(A-->G) and nt--29(A- >G) mutations were six (8.2%) and four (5.5%) respectively. PMID- 8732053 TI - [Anti-metastasis gene nm23 expression in buccal cancer studied by immunohistochemistry stain]. AB - The nm23 gene has been proposed as a candidate metastasis-suppressor gene. In this study we used immunohistochemistry stain for polyclone antibody nm23 in 62 cases of buccal carcinoma. The results showed that both the epithelium of mucosa and the duct epithelium of minor salivary gland nearby the tumor were strong positive. The stains on tumor cells were various. 31.0% (9/29) of the cases with lymph-node metastasis were positive while 57.6% (19/33) of those with no lymph node metastasis positive (P = 0.044). Histologically, 59.4% (19/32) of the cases were positive in Grade I, while 37.5% (6/16) positive in Grade II and 21.4% (3/14) in Grade III (P = 0.048). The positive rate increased with the patients survival time from less than 3 years to 3-5 years and more than 5 years correspondingly, but there was no statistical significance (P = 0.126). The results suggested that the expression of nm23 might be one of the factors influencing the lymph-node metastasis and prognosis of buccal cancer. PMID- 8732054 TI - [Synthesis of substituted benzeneseleninic acid and its antineoplastic activity]. AB - In view of the negative relationship between selenium and cancer, we designed and synthesized eleven substituted benzeneselenic acids (IVa-k) The antineoplastic activity of the title compounds was evaluated via the tests of inhibition of acetylcholinesterase, HL-60 and K562 in vitro, and Ehrlich carcinoma in mice. The result showed that the compounds IVa and IVb had distinct antineoplastic activity. The life span increase rate of IVb on mice bearing Ehrlich carcinoma was 20.30% (20mg/kg), which was higher than the increase rate of sodium selenite (10.65%, 0.5mg/kg). PMID- 8732055 TI - [Effect of tetrandrine on pulmonary hypertension induced by monocrotaline in rats]. AB - The effects of Tetrandrine (Tet) on intraacinous pulmonary arteries (IAPA) and bemodynamics were studied by means of a rat pulmonary hypertension model induced by monocrotaline. The results showed that Tet could reduce the contractive pressure of pulmonary artery, right ventricle and right atrium of heart by 28.0%, 28.3%, and 31.5% respectively. It could also decrease the vascular degeneration of endoepithelial cells of IAPA, the endoepithelial subcavity, the collagens of medial membrane and the numbers of smooth muscles of IAPA. PMID- 8732056 TI - [The effects of Salvia miltiorrhiza and polysaccharide sulphate on the adhesion of erythrocytes of the patients with cerebral thrombosis to cultured endothelial cells]. AB - Salvia miltiorrhiza and polysaccharide sulphate are commonly prescribed for curing cerebral vascular diseases. In this study, we found that the adhesion of erythrocytes of the patients with cerebral thrombosis to cultured human umbilical vein endothelial cells was decreased in number and intensity after the erythrocytes were treated with salvia miltiorrhiza and polysaccharide sulphate in flow field. Moreover we found that polysaccharide sulphate was more effective than salvia miltiorrhiza in preventing adherence in common doses. The two drugs' effects of preventing adherence might be an important mechanism for curing the patients with cerebral thrombosis. PMID- 8732057 TI - [Effects of ginkgolide B on isobaric hypoxic pulmonary hypertension in rats]. AB - We examined the effects of ginkgolide B (BN52021), a specific platelet activating factor (PAF) antagonist, on hypoxia-induced pulmonary hypertension and pulmonary vascular remodeling. Chronic hypoxia can cause pulmonary hypertension and pulmonary vascular remodeling in rats. Treated with BN52021, the rat's pulmonary hypertension, pulmonary vascular remodeling and right ventricular hypertrophy at 3 weeks of hypoxic exposure (FiO2 = 0,10) could be reduced. The results suggest that PAF contributes to the development of chronic hypoxic pulmonary hypertension and pulmonary vascular remodeling, and PAF antagonist may be an useful agent for preventing hypoxic pulmonary hypertension. PMID- 8732058 TI - [153Sm-EDTMP for moderate and severe bone cancer pain]. AB - One hundred and thirty-six patients with bone cancer pain were treated with 153Sm EDTMP (ethylenediamine-tetramethylene phosphonic acid). Pain free was noted in 49 cases (36%, 49/136) and pain relief in 77 cases (56.6%, 77/136), the total relief rate being 92.6% (126/136). The data from 76 patients with moderate and severe pain showed there were no significant relationships between the patients' age, the dose of 153Sm-EDTMP and the analgesic effects (P > 0.05). The pain relief observed in the patients with chest pain (ribs metastases) was earlier than that in other groups (P < 0.05). We didn't find any clinical side-effects, so 153Sm EDTMP is safe for use. PMID- 8732059 TI - [Detection of serum isoferritin level in 195 patients with cancer]. AB - ELISA (double-determinant) was used to detect the serum isoferritin (SIF) levels in 42 patients with nasopharynx cancer (NPC), 38 with lung cancer (LC), 26 with esophageal cancer (EC) and 89 with other cancers, a total of 195 cases (151 males and 44 females). The results revealed that the positive rate of SIF in patients with cancer was 73.8% (144/195), among which, the positive rates in patients of stage I-II, stage III-IV, in patients not operated upon and in those after operation were 64.8%, 81.3%, 81.4% and 63.4%, respectively. Statistics was significantly different in SIF level between the stage I-II and III-IV groups, and between the un-operation and after operation groups (P < 0.001) by rank sum testing. The positive rates of NPC, LC and EC were 73.8%, 86.8% and 73.1%, respectively. The results have demonstrated that isoferritin is a malignant neoplasm associated antigen, and SIF level is related closely with the development of the tumor and operative treatment. PMID- 8732060 TI - [The sandwich enzyme-linked immunoabsorbent assay of serum transferrin receptor by using monoclonal and polyclonal antibodies]. AB - The human placenta transferrin receptor was purified in the form of transferrin transferrin receptor complex (Tf-TfR), and a monospecific polyclonal antibody against TfR was developed by a Tf-coupled Sepharose 4B affinity chromatography to remove the anti-Tf components in the antiserum. A sandwich enzyme-linked immunoabsorbent assay (ELISA) was established for measuring serum transferrin receptor (sTfR) by using monoclonal antibody OKT9 and monospecific polyclonal antibody. This method is simple, specific and sensitive and has a good accuracy. The measurement of sTfR showed that the level of normal children was 4.54 +/- 1.08 mg/L. There were increased levels of sTfR in patients with severe iron deficiency anemia and those with hemolytic anemia (13.92 +/- 4.45 mg/L and 9.94 +/- 3.22 mg/L, respectively). In patients with aplastic anemia, the level was decreased (2.06 +/- 0.82 mg/L). These results indicate that the sTfR measurement has a differential significance for diagnoses of various anemia. PMID- 8732061 TI - [Effects of vesnarinone on L-type single Ca2+ channel current in cultured ventricular myocytes of embryonic chick]. AB - The effects of vesnarinone on L-type Ca2+ channel current of embryonic chick ventricular myocytes were studied with cell-attached patch clamp technique. Vesnarinone was used in the concentrations of 10(-5), 2 x 10(-5) and 10(-4) mol/L; the availability (P8) of Ca2+ channel activity was increased (P < 0.001) markedly (about 2-3 fold). The increasing of the open probability (Po) of Ca2+ channel by vesnarinone showed concentration-dependence. The increase was 2.25 (10(-5)mol/L), 3(2 x 10(-5)mol/L) and 4.3(10(-4)mol/L) fold. The open time rate constant (tau 0) and close time rate constant (tau 1 and tau 2) of Ca2+ channel were not changed in the concentrations of 10(-5) and 2 x 10(-5) mol/L, but the tau 0 was slightly decreased in the concentration of 10(-4) mol/L. The actions of vesnarinone on single Ca2+ current were compared with those of milrinone (it is known as an PDE inhibitor which had inotropic effect associated with an increase in cytoplasmic cAMP level.) The effects of milrinone on P8, tau 0 and tau 1 were similar with those of vesnarinone (10(-4) mol/L), but the increase of Po was 2 fold less than that of vesnarinone. The results showed that apart from the role of PDE inhibitor, vesnarinone has other effects on myocytes. PMID- 8732062 TI - [A study on the effective way and dosage of intravas deferens injection of HFMC for male contraception]. AB - To investigate the effective way and dosage of intravas deferens injection of HFMC for male contraception, Teflon tubes of 1 mm caliber were used to imitate intravas injection of HFMC. We injected HFMC first, then the precipitant; thus HFMC could adhere to the walls and blend with the precipitant to precipitate. Eight adult male rabbits, exposed reproductive ducts were injected with dyed HFMC in opposite directions (upward and downward), HFMC flowed out of the distal end of the vas easily when its volume was more than that of the vas. 14 adult rabbits were used for the comparison of sperm density and fertility index between upward and downward injections of HFMC, and between the dosage of HFMC (50 microliters and 100 microliters) injected upward. The results show that it is difficult to attain contraception by intravas injection of HFMC downward, but contraception is quite possible by intravas injection of HFMC upward providing the volume is enough. The result also suggest that HFMC should be injected into vas deferens upward first; its volume should be 3 time as much as the natural volume of the vas from the injecting point to the epididymal end. Then a little precipitant is injected into the vas. PMID- 8732063 TI - [Ultrastructural study of human yolk sac]. AB - Twenty cases of human yolk sac of different gestation weeks observed by electron microscopy. The wall of yolk sac consisted of endoderm, mesenchyme and mesothelium. Nuclei of the endodermal cell were large and nucleoli were remarkable. The surface of endodermal cell presented a number of microvilli. There were Golgi complex, rough endoplasmic reticulum, glycogen and mitochondria in the cytoplasm of the endodermal cell. The fibroblast-like cell, macrophage like cell. erythroblast in various mature degree, collagen filament and capillaries were found in the mesenchyme. The mesothelial cell had long, slender microvilli, which were mostly branched. Pinocytotic vesicles in the apical cytoplasm beneath the mesothelial cell membrane were frequent. The differently sized endodermal tubules with well-defined lumen and microvilli were recognized by electron microscopy. Masses of endodermal cells frequently invaginated in the mesenchymal layer, and they often reached the mesothelial layer. These structures of yolk sac were the foundation of finishing synthesis, secretion, translation and hematopoiesis during early organogenesis. PMID- 8732064 TI - [The establishment of ADBS data bank system for autopsy records]. AB - The ADBS data bank system consists of PC 286 microcomputer, (or 386 and other more advanced machine) VGA color monitor, printer and ADBS V. 2.0 software. According to the international classification of diseases (ICD-9 or ICD-10, SNOMED), pathological diagnoses are coded and put into computer. This system has five functions: search, statistics, edition, print and system restore. For facilitating the operation of this system, all orders are listed as window menu. The statistic results can be shown as tables or figures and put out as disk files or prints. The system is currently an advanced computerized system of histopathological and autopsied records in Chinese. PMID- 8732065 TI - [Immunohistochemical study of 5-HT on gunshot wound of human skin]. AB - Fourty antemortem and 30 postmortem gunshot wound samples of human skin were studied by immunohistochemical method. The 5-HT was seen mainly in wound edge, papillary layer, hypodermis and surrounding tissue of all antemortem gunshot wounds. The 5-HT was also discovered on the postmortem gunshot wounds which occurred within 8 minutes after death. The result showed that immunohistochemical staining of 5-HT be useful for diagnosing the antemortem gunshot wound. It also demonstrated that the mast cells of human do not contain 5-HT. We also studied the rate of mast cell degranulation of gunshot wounds of human skin by toluidine blue staining. The rate of mast cell degranulation of antemortem gunshot wounds increased (50%), and it was higher than that of postmortem gunshot wounds, suggesting that the rising of degranulation rate is a sign of antemortem injury. PMID- 8732066 TI - [Genetic polymorphism of properdin factor B in the Han population in Chengdu, China]. AB - The genetic polymorphism of properdin factor B(Bf) was first investigated in the Han population in Chengdu by isoelectrofocusing and immunofixation. The results revealed BfSS (129 cases), BfFS (48 cases) and BfFF (9 cases). No Bf variants were noted in this study. The allele frequencies were: Bf * S 0.8226, Bf * F 0.1774. PMID- 8732067 TI - [Effect of olbetam on hyperlipidemia in NIDDM]. AB - The effect of Olbetam on serum lipid and lipoproteins was studied in 30 diabetic patients with hyperlipidemia in four weeks trial. The dose of Olbetram was 500 mg/d. The results showed serum concentrations of TC, TG, and VLDL-C were decreased while HDL-C especially HDL2-C increased significantly after treatment. There were no significant changes in FBG, blood creatinine and urine acid. This result suggests Olbetam can improve dyslipidemia in NIDDM and was well tolerated by all patients. PMID- 8732068 TI - [A preliminary observation of preventive and blocking effect of Gynostemma pentaphyllum (Thunb) Makino on esophageal cancer in rats]. AB - Gynostemma pentaphyllum (Thunb) Makino is a kind of natural plant growing in the southern provinces of China. In this experiment, rats were given drinks of 2% Gynostemma Pentaphyllum (Thunb) makino aqua boiled solution freely for 2 weeks before administration of carcinogen. Subsequently, the rats were injected with carcinogen (MANA) for 18 weeks and were sacrificed at intervals. The result showed that the number of cases of esophageal epithelial papilloma, the average number of tumor and the incidence of esophageal cancer in the experimental group were lower than those in the control group (MANA only), though no statistical significant difference was found. The initiation of cancer was delayed for 6 weeks. These indicated that Gynostemma Pentaphllum Makino might have some preventive and blocking effect on esophageal cancer in rats. PMID- 8732069 TI - [Toxicity of vanadium to Leydig cells in vitro]. AB - The levels of testosterone secreted by rat's Leydig cells in varied concentrations of V2O5 (0.125, 0.,25, 0.5, 2 and 3 mmol/L) were assayed. The result showed there was no obvious relation between the level of testosterone and the concentration of V2O5. This in vitro study in accord with the result of whole animal research indicates that Leydig cell is not the target of V2O5. PMID- 8732070 TI - [Identification of streptavidin]. AB - In this report, streptavidin (SA) was further identified on the basis of our previous researches in SA isolation and purification. 23.2 mg of SA productivity could be obtained after purification by DEAE-52 column from 1 L of culture supernatant. The spectrum scanning of ultraviolet absorption was congruent with that of the Sigma product. 2 bands appeared on the SDS-PAGE electrophoresis of SA. The molecular weight of SA was 72 881 Dalton with PI 6.0; the averaged biotin binding activity was 18 micrograms/mg. The results confirmed that the methods of isolation and purification of SA developed by authors were reliable for immunological application. PMID- 8732071 TI - [Determination of paracetmol caffeine and ephedrine hydrochloride in likeshu capsules]. AB - Paracetamol (P), caffeine (C) and ephedrine hydrochloride (EH) are there physiological active component in Likeshu capsules. We have proposed the determination method of these three components using extractive separation spectrophotometry. Only one amount weighed of sample is required to determine them. The recoveries of P, C and EH were 97.51 +/- 0.64% (n = 9), 98.24 +/- 1.04% (n = 8) and 98.58 +/- 1.09% (n = 9) respectively. The results of determination of samples agreed with those of the HPLC method. This method is sensitive, accurate and simple. The other ingredients of Likeshu capsules do not interfere. PMID- 8732072 TI - [A study on the simultaneous HPLC determination of chlorhexidine and its impurity 4-chloroaniline]. AB - This paper reports a new RP-HPLC method using phenacetin internal standard (IS) for simultaneous determination of chlorhexidine (CH) and its impurity 4 chloroaniline (4-CA). We use the LC-3A HPLC instrument, a Zorbax-C8 column (25cm x 4.6mm id, 10 microns), a variable wave length UV detector and C-RIA chromatographic data processor. A mixture of methanol and 0.2 mol/L NaH2PO4 (50.5 : 49.5, pH 3.0) is used as mobile phase, whose flow rate is 1.0 ml/min. Sample solution of 15 microliters is injected onto the column, which is maintained at 30 degrees C. The eluent is monitored at 240nm. The retention times are 16.72 min for CH and 5.54 min for 4-CA. The standard curves of CH and 4-CA are established using peak area ratio of CH/IS and 4-CA/IS to concentration respectively. They are both linear (r = 0.9999) within 15-200 micrograms/ml for CH and 60-1200 micrograms/ml for 4-CH. The limits of detection are 1.33 micrograms/ml for CH AND 7.5 micrograms/ml for 4-CA (S/N = 3). The analytical recovery rates are 100.1% for CH and 103.1% for 4-CA. This method has been used for investigating the stability of CH preparation. PMID- 8732073 TI - [Concentrations and size distributions of aerosols in middle summer of Chengdu]. AB - Statistical concentrations and size distributions of aerosols in middle summer of Chengdu city were measured by using optical scattering method. The influence of climate and its daily change affects on the concentration and distribution were analysed. The results showed: many of the distributions agreed with the Junge distribution; the peak concentration appeared near the noon; the relative humidity had strong influence on concentration; the clean-up effect of rain fall was evident. The measured results are useful for evaluating the environmental sanitation of the city. PMID- 8732074 TI - [Experimental studies on the preventive effect of artemether against schistosomal infection]. AB - In order to study the preventive effect of artemether (Art) against schistosomal infection, mice, rabbits and dogs infected with Schistosoma japonicum cercariae were treated intragastrically (ig) with Art on various days after infection. The preventive effect was assessed by total and female worm reduction rate, the number of the animals without female worm, measurement of some parameters relevant to the acute schistosomiasis, and histopathological observation on the liver. The results showed that in mice a promising effect was obtained when an initial dose of Art 300 mg/kg was given ig on d7 after infection with cercariae, followed by repeated dosing every wk for 4 times. In rabbits and dogs treated ig with Art according to above-mentioned appropriate regimen on d7 after infection at a dosage of 10 or 15 mg/kg given every 1 or 2 wk for 2-4 times, the total and female worm reduction rates were 96.8%-100%, and part of the animals were free from female worm. After preventive treatment of the rabbits with Art, the body temperature and eosinophil count were normal and no schistosome egg was found in the feces. When rabbits infected with 48-52 schistosome cercariae once every other day for 5 times were initially treated ig with Art 15 mg/kg on d7 after the first infection, and the same dosing was given repeatedly every 1 or 2 wk for 2-4 times, the total and female worm reduction rates were 97.6%-98.4%. Histopathological observation showed that the above-mentioned infected dogs and rabbits received preventive treatment with Art exhibited a promising protective effect on their livers. The protective effect of Art given every 1 wk on the liver was similar to that of the drug given every 2 wk. The results indicate that Art possesses an effect for prevention of schistosomiasis which could be helpful for reducing infection rate and intensity of infection, and controlling acute schistosomiasis. PMID- 8732075 TI - [The sensitizing role of the anti-idiotypic monoclonal antibody-NP30 of Schistosoma japonicum in egg granuloma formation]. AB - C57BL/6 mice sensitized intraperitoneally with the anti-idiotypic monoclonal antibody-NP30 of Schistosoma japonicum and then injected with vital egg suspension directly into spleen were used as the experimental models of hepatic egg granulomas in this study. The soluble egg antigen(SEA) sensitized group was used as positive control and the SP2/0 ascites injected group as negative control. The results showed that the size and volume of hepatic egg granulomas in the NP30 sensitized group increased obviously 4 days after egg injection and that a large number of eosinophiles appeared in egg granulomas. The size and volume of egg granulomas reached the peak values at d8 and d15 after egg injection, respectively. The peak values of the NP30 sensitized group were similar to those of the SEA sensitized group (P > 0.05). The size and volume of egg granulomas in the SP2/0 ascites injected group did not reach the peak values, which were less than those of the group sensitized with NP30 (size: P < 0.05; volume: P < 0.01), until d32 after egg injection. Our study suggests that NP30 may play a sensitizing role in the formation of hepatic egg granulomas. PMID- 8732076 TI - [A study on behavioural characteristics of staying on the mountain and its relationship with malaria infection in Li and Miao minorities in Hainan Province]. AB - A study of sociology combined with epidemiology was conducted in Li Minority's two villages and Miao Minority's two villages nearby the foot of mountain in a historically malaria hyperendemic area, Nanqiao Township of Wanning County in August, 1993. The results showed that malaria infection was closely correlated to the behaviour of staying overnight on the mountain of Li and Miao Minorities during farming seasons. Most of Li people stayed overnight on the mountain for planting areca were adults, accounting for 21.6% of the whole population, the duration of stay was about half a year; Miao people stayed overnight on the mountain planting and harvesting upland rice, always with their family member, so the percentage of people who had stayed on the mountain was as high as 82.1%, the duration of stay was about one and half a month. Blood smear examination showed that the malaria parasite rates in both Li and Miao villages were 11.1% and 24.1%, respectively, the positive rate of P. falciparum was 1.0% and 9.1%, respectively, the positive rate of IFA (titer >or= 1 : 40) was 31.2% and 46.5%, respectively. It is indicated that both the rate of parasitaemia and the P. falciparum infection were much higher in Miao Minority than those in Li Minority (P < 0.001). A cohort analysis showed that the malaria parasite rate of population who having not stayed, having ever stayed and being stayed on the mountain was 6.5%, 27.4% and 42.1%, respectively, of them the rate of P. falciparum infection was 0.5%, 8.8% and 18.4%, respectively. The results suggest that malaria acquired from mountainous forest referring to "up-mountain infection" is a major source of malaria infection and a significant risk factor in determining the prevalence of malaria (especially falciparum malaria) in Hainan Province. This behavioural risk factor has become the main obstacle for malaria control in Hainan Province. Therefore, controlling "up-mountain infection" and adopting appropriate anti-malaria measures to protect the risk population who have to work and stay on the mountain should be strengthened in the malaria control program. PMID- 8732077 TI - [Erythrocyte immunity and regulating factors in mice infected with Plasmodium berghei ANKA strain]. AB - After seventy-two ICR mice were inoculated with Plasmodium berghei ANKA strain, parasitaemia was revealed in all animals inoculated in two to seven days. During the course the number of malaria parasites increased rapidly from 2.3 +/- 1.3 x 10(3) on d2 to 93.7 +/- 1.8 x 10(3) on d7, the number of erythrocytes increased from 8.2 +/- 0.9 x 10(12)/L on d0 to 11.1 +/- 1.0 x 10(12)/L on d2 and then decreased gradually, reaching 1.9 +/- 0.4 x 10(12)/L on d7, and the number of white blood cells appeared to be fluctuated. Additionally, the rosette rate of erythrocyte=C3b receptor and the rosette rate of erythrocyte-immunocomplex increased on d2 and then decreased gradually to a very low level, suggesting that the erythrocyte immune function reduced in malaria. It is assumed that the reduction in erythrocyte immune function in malaria is attributed to the damage of the erythrocyte membrane in addition to the changes in the function of the erythrocyte immunoregulating factors. PMID- 8732078 TI - [Status and strategy of parasitosis control in preventive health care in China]. AB - According to the result of nationwide human parasitic disease distribution survey, the public significance of parasitosis control was set forth in terms of preventive health care and economic impact as well as parasitosis control and primary health care. A suggestion was made on the control strategy aiming at the parasitosis of priorities listed in the above survey. PMID- 8732079 TI - [Relationship between the rate of parasitic infection and the knowledge of prevention]. AB - The relationship between the rate of parasitic infection and the level of knowledge of prevention was studied among 5479 persons allocated to 12 randomly selected groups. Stools were examined for parasitic infection by using Kato-Katz, iodine direct smear amd tube culturing methods. Children under 15 were examined for Enterobius infection by cellophane tape method. Individuals were considered infected when any parasite species was found in the stool and the infection rate was computed. The score on knowledge of prevention was measured by devising a questionnaire comprising 11 questions on parasitic disease transmission, clinical signs and prevention methods. Ten marks were given for each question correctly answered. The result of the investigation showed that the total infection rate was 52.2% and there was no significant difference between males and females (X2 = 1.94, P > 0.05). The infection rate increased with a decrease in the level of education and the highest was found in 5-year-old age group. There were altogether 7 species of parasites detected in all age groups. The infection rate of Ascaris lumbricoides was 31.3%; Trichuris trichiura, 34.1%; hookworm, 5.3%; Enterobius vermicularis, 20.3%(227+1 117); Fasciolopsis buski, 0.8%; Entamoeba coli, 0.8%; Giardia lamblia, 0.2%. The average score on knowledge was 41.9, being significantly higher in males than in females. Analysis for the age distribution showed that the highest score was found in 15-year-old age group. The data of infection rate of different parasites were converted by arcsine transformation and compared with the average score, showing that there were negative relations between the average score and the infection rate of Trichuris, hookwork, Entamoeba and the total incidence. The value of r was --0. 5534, --9.5557, - 0.6091 and --0.604. respectively. The number of parasite species infecting individuals was also found to have negative relation to the score on knowledge of prevention. PMID- 8732080 TI - [Studies on the bionomics of Phlebotomus chinensis in mountain cave and its relation to control in northern Sichuan]. AB - After a survey of the gonotrophic cycle of 5147 female Phlebotomus chinensis collected from the caves in the mountain area 1500 m above sea level in Nanping County, Sichuan Province where this sandfly was proved to be the typical predominant wild species living mostly in the wild cave at a very high density, a large cave was sprayed with alphamethrin at a dosage of 50 mg/m2 in July, 1993. As a result, the sandfly density dropped to zero spanning over a period of two peak seasons. The authors consider that the residual effect of alphamethrin as long enough to suppress sandfly density in the cave for at least two years, and stress that the spray is a favourable measure in all the year round in reducing the sandfly population density and preventing visceral leishmaniasis. PMID- 8732081 TI - Collaborative study on detection of circulating antigens in schistosomiasis japonica. AB - A workshop for collaborative study on circulating Schistosoma japonicum antigen detection was held at the Hunan Institute of Parasitic Diseases, Yueyang, Hunan Province in May, 1993. Eight laboratories from schistosomiasis-endemic provinces (municipalities) were joined in this study. Scientists from these laboratories performed serological assays in blind using their own brought test systems that developed during recent years. The serum samples tested covered 100 non-endemic controls, 190 from patients with active schistosomiasis, 110 from cured cases of schistosome-infection, and 170 from individuals harbouring other parasitic infections or hepatitis. Results of the study revealed an assay specificity, in terms of false positive rate, of 2% in Series B versus 14-46% in the other 7 series. With regard to assay sensitivity, all test series showed 95-100% for acute cases. While for chronic cases, test series A and F demonstrated sensitivities at 73.3% and 72.7% respectively. Other series were all below 70%. Negative reversion rates for sera from cured cases within 1 year post-treatment were below 33.3%, those within 2 years were 60% for Series E, while below 50% for all others except the Series H which was excluded due to an extremely low sensitivity obtained. The low values of these test series for chemotherapy assessment may be attributed to the poor specificity of the detecting systems used and/or the sample donors being not totally cured. It is strongly indicated that all the 8 test systems involved in the present study should further be improved in sensitivity, specificity as well as in capacity for pre-and post treatment evaluations. PMID- 8732082 TI - Effect of monoclonal antibodies on the entry and development of Plasmodium vivax sporozoite in cultured cells. AB - We have observed the effect of monoclonal antibodies against sporozoites (2F2,NV3,2E10) and blood stages (4B2,8E3) of P. vivax on entry and subsequent development of P. vivax sporozoites in HepG2--A16 cells in vitro. The results demonstrated that inhibitory effects of monoclonal antibodies on the attachment and entry were found to be related to the antibody concentration. At 25 micrograms/ml, the percentages of inhibition of sporozoite invasion were 100% (2F2), 76%(NVS3) and 10.5% (2E10). Even if the invasion was not totally inhibited, the presence of abnormal exoerythrocytic schizonts suggested that continued effect of antibodies after sporozoite penetration still existed. No significant effects of 4B2 (4.5%) and 8E3 (3.4%) were recorded as compared with normal mouse serum. These findings indicate that inhibition of sporozoite invasion assay may be useful for determining the protective effect of anti sporozoite antibodies in vitro. PMID- 8732083 TI - [Scanning electron microscope observation on the eggs of Musca domestica vicina]. AB - The surface structures of the eggs of house fly (M. domestica vicina) were examined by SEM. A layer of membranaous structure with small holes in clusters covering the whole egg was seen. The membranous structure is supported by large amount of tissue masses of different sizes. The submembranous tissue is dotted with numerous concave-holes. A hatching crack with a layer of very thin and vulnerable membrane runs lengthwise across the concave surface of the egg. PMID- 8732084 TI - [Comparative study on hepatic and intestinal pathological changes in different animals with schistosomiasis japonica]. AB - The pathological changes of livers and intestines were compared among cats, dogs and rabbits infected with Schistosoma japonicum cercariae for three months. The resuults showed that the most serious lesions were seen in rabbits while the slightest were in cats. The granulomatous response was more distinct in livers than in intestines in all the three animals. The increment ratio of egg granuloma in livers was 3.52, 2.64 and 1.81 in rabbits, dogs and cats, respectively. The eggs were deposited singly or in strings, few granuloma formation was observed in the intestines. In addition to inflammatory infiltration, swelling and/or dropping of mucosa the gland destruction and ulcer were also observed in the mucosa. The investigation demonstrated that pathological lesions varied in different animal species. PMID- 8732085 TI - [Sarcocystosis]. PMID- 8732086 TI - [t(14:18) chromosomal translocation and over expression of bcl-2 protein in follicular type non-Hodgkin's lymphoma]. AB - Polymerase chain reaction (PCR) and immunohistochemical staining methods were used to detect the bcl-2/IgH fusion gene [t(14:18)] and the overexpression of bcl 2 protein in 28 cases of follicular lymphomas and 18 cases of reactive follicular hyperplasia. The results showed that the bcl-2/IgH fusion gene was present in 57% of follicular lymphomas and that this translocation was also present in 33% of the reactive follicular hyperplasia cases. The over expression of bcl-2 protein was found in 78% of follicular lymphomas, but none in follicular hyperplasia, therefore bcl-2 protein staining may be a useful immunohistochemical method to differentiate follicular lymphoma and follicular hyperplasia. PMID- 8732088 TI - [Gene therapy of human pancreatic carcinoma by recombinant retroviral vector expressing herpes simplex virus thymidine kinase gene]. AB - A recombinant retroviral vector pNTK expressing the "suicide" gene herpes simplex virus thymidine kinase (HSV-TK) gene was constructed. The recombinant vector was packaged by PA317 cells and viral supernatant was used to infect the human pancreatic carcinoma cell line PC-2. After selection in G418, resistant colonies were identified and isolated. Cytotoxic effects of the non-toxic prodrug Acyclovir (ACV) or Ganciclovir (GCV) to the NTK transformant PC-2 cells was more than 90%, while that of the control PC-2 cells was less that 10%. Furthermore, the "bystander effect" of HSV-TK on PC-2 cells was also observed. PMID- 8732087 TI - [Hepatitis B virus infection and pathogenesis of glomerulonephritis]. AB - In order to investigate the role of hepatitis B virus (HBV) infection in the pathogenesis of HBV associated glomerulonephritis, 50 cases of glomerulonephritis with positive HBV infection marker antigenemia and/or HBAg detected by immunohistochemistry in renal tissue were collected. The distribution and localization of HBV DNA were observed by using in situ hybridization. In addition, Southern blot analysis was performed in 23 of the 50 cases to reveal the state of renal HBV DNA. Thirty Six cases (72%) were found to be HBV DNA positive by in situ hybridization, which was localized in the nucleus of tubular cells. In 26 cases HBV DNA was simultaneously detected in the nucleus of glomerular mesangial and epithelial cells as well as in the mesangial matrix. Seventeen of the 23 cases proved to be HBV DNA positive by Southern blot analysis. In which 3 cases were identified as having non-replicating free HBV DNA, while 14 cases (82%) were of the integrated form. Since there was abundant evidence of the renal tissue being infected with HBV, it was considered that the HBAg deposited on glomeruli not only originated from circulation but also from the HBV infected glomerular cells. In addition to the humoral immune injury mediated by HBAg-HBAb immune complex, the cellular mechanism mediated by target antigen (HBcAg) may be also involved in the pathogenesis of HBV associated glomerulonephritis. PMID- 8732089 TI - [The reversing effect of HPV-16 E6E7 gene antisense plasmid on the HPV-16 positive human cervical carcinoma cells]. AB - Antisense reconstruct, p16as E6E7Neo, harboring HPV-16E6E7 gene of which the expression is regulatable by dexamethasone was constructed. Calcium phosphate mediated transfection was employed to transduce this antisense reconstruct into HPV-16 positive CasKi cell line and HPV negative C-33A cell line (both derived from human cervical carcinoma) respectively. After dexamethasone was added into the culture medium to induce the expression of E6E7 antisense gene, the malignity of CasKi cells were reversed, while the growth characteristics and malignity of C 33A cells were unchanged. The results show that the reversing effect on CasKi cell is specific and mediated by inhibition of expression of E6E7 gene. It also demonstrated a more profound pathogenic role of HPV-16 E6E7 gene in the tumorogenesis of cervical carcinoma and provides a possibility for gene therapy of this tumor. PMID- 8732090 TI - [Detection of the expression of P21, P53, P185 proteins and the mutation of ras, p53 genes in colorectal adenoma and carcinoma]. AB - Expression of P21, P53, P185 proteins, mutations of ras, p53 genes in colorectal adenoma, carcinoma and transitional mucosa were studied using immunohistochemicstry and PCR-RFLP methods. The results showed that the positive rates of P21, P53 and P185 proteins in colorectal adenoma were 53.3%, 27.6% and 13.3% respectively, the expression of P21 and P53 were associated with the malignant potential of adenoma. The positive rates of P21, P53 and P185 proteins in colorectal carcinoma were 72.9%, 37.8% and 47.2% respectively. 9 adenomas and 40 carcinomas contained more than two protein expressions and their co-expression was associated with the malignant potential of adenoma and the prognosis of carcinoma. The mutation rates of ras gene in colorectal adenoma and carcinoma were 26.7% and 41.9% respectively. The ras gene mutation was associated with the malignant potential of adenoma. The mutation rates of p53 gene (codon 248) in adenoma and carcinoma were 3.3% and 14.9% respectively. The prognosis of patients having gene mutation of both ras and p53 were poor. The results suggested that the alterations of ras, p53 and c-erbB-2 genes are involved in the tumorigenesis and development of colorectal carcinoma. PMID- 8732091 TI - [nm23 gene product/NDPK expression and its clinical significance in human colorectal carcinoma]. AB - The expression of nm23 gene product/NDPK was examined immunohistochemically in resected human colorectal carcinomas using anti-NDPK polyclonal antibody. Of the 103 carcinomas tested, 76.7% (79 cases) showed positive staining of nm23/NDPK in the primary lesions. In patients without regional lymph node metastasis, the positive staining rate (90.5%, 38/42) was significantly higher than that in those with nodal involvement (67.2%, 41/61). Inverse relationships were found between nm23/NDPK expression and tumor invasion or the progression of Dukes' staging. Expression of NDPK was positively associated with longer post-operative survival and the relationship was significant. Therefore, our data indicated that nm23 gene plays an important role in lymph node metastasis and invasion in colorectal carcinomas. The expression of nm23 gene product/NDPK may be a useful tool for prognosis of human colorectal carcinomas. PMID- 8732092 TI - [Missed diagnosis in prostate incidental carcinoma]. AB - 522 cases of benign prostate hyperplasia from 1980 to 1994 were reviewed. Prostate incidental carcinoma (PIC) was identified in 26 cases (4.98%). In order to study the pathomorphological features and to study the causes for missed diagnosis of this entity, the authors observed and analysed the numbers of carcinomatous foci their distribution, classification, staging and differentiation, morphological features and prostate specific antigen (PSA) markers. It was found that 23 cases exhibited a pathological structure of uniform pattern and 3 cases with pluriform pattern. Adenocarcinoma was most common. 17 cases were A1 stage and 9 cases were A2 stage. Isolated multiple foci of invasion and alterations of architecture are important pathological features of these cases. Missed diagnosis of prostate cancer is related to the small amount of biopsy tissue, contrast between benign and malignant prostate epithelial tissue not distinct, scarce nuclear anaplasia, careless microscopic observation, or observer lacks the ability to differentiate between pathologic types and pluriform patterns. Different PSA reactions may be related to tumor differentiation and classification. PMID- 8732093 TI - [A clinicopathological study of 3 dimensional structure in adenoma of large intestine]. AB - The 3 dimensional reconstruction electrocomputor was used to study the distribution of atypia epithelia in 50 cases of large intestinal adenoma which were taken by fibercoloscopic resection. The 0.2 mm interval serial sections were stained with HE. Carcinomatous changes in adenomas were found in 17 cases, the rate being 34%. The average size of intraadenoma carcinoma being 3.4 times that of adenoma. The larger the adenoma, the higher the cancer rate. But the adenoma to cancer rate is high (25%) in Ips type adenoma of small size with pedicle. The size of adenoma is not associated with allotype incidence. 14 of the 17 cases were detected by study of conventional paraffin block slides, missed diagnosis rate being 18%. The serial sections must be made with intervals less than 0.6 mm in order to prevent missed diagnosis. This method is also of significance to determine if the excision site is cancer free. PMID- 8732094 TI - [An immunohistochemical study of intestinal mucinous antigens and CEA in ovarian epithelial tumors]. AB - Using monoclonal antibodies specific to the large intestinal mucinous antigen (LIMA), small intestinal mucinous antigen (SIMA) and CEA, the expression and localization of these proteins in 78 formalin fixed paraffin embedded tissues from ovarian epithelial tumors were studied. The localization of LIMA, SIMA and CEA was associated with tumor features and differenciation. Positive stain of benign and borderline tumors were mostly located on the lumina border while malignancies were situated both on the lumina border and in the cytoplasm and tended to concentrate in the latter with the increase in histological grade. No difference in positive rate was seen between serous and mucinous carcinomas. However, staining intensity of CEA in serous carcinomas was lower than that in mucinous carcinomas and the expression and intensity in endometrioid carcinomas were low. On the other hand, positive CEA and LIMA was associated with a significantly worse survival. PMID- 8732095 TI - [Clinicopathologic analysis of ependymoma in childhood]. AB - A review of the clinicopathologic features of 23 ependymomas in children was made, of which 17 were male and 6 female, the male:female ratio being 2.8:1. Their average age was 8.8 years. Seven of the 23 ependymomas were supratentorial, 16 were infratentorial. The mean duration of symptoms prior to operation in the two groups was 5.3 and 3 months respectively. Histologically, the tumor could be classified into three types: papillary (or myxopapillary), epithelial and cellular. According to the degree of differentiation, the tumor could be divided into typical ependymoma and anaplastic ependymoma. Immunohistochemically, 17 of the 23 cases were positive for glial fiber acid protein (GFAP) and the remaining 6 cases were negative. The clinicopathologic features of childhood ependymomas, factors influencing prognosis and histologic classification are discussed. PMID- 8732096 TI - [Gamma interferon-like immunoreactivity in adrenal gland and cortical tumor]. AB - Interferon-gamma (IFN-gamma) has many important immunoregulatory functions. It has previously been presumed to be produced by activated lymphoid cells alone. Recently, IFN-gamma like immunoreactivity has been described in certain parts of the nervous system and the skeletal or cardiac muscle system in the rat. We wanted to know whether this molecule could be expressed in the human adrenal gland and cortical tumors. Adrenal tissue samples were taken adjacent to 14 tumor cases, from 17 adrenal cortical adenomas, and 7 cortical carcinoma cases. Deparaffinized 5 microns sections were stained with IFN-gamma monoclonal DB-1 antiserum. The avidin-biotin complex method was used. IFN-gamma appeared in the cytoplasm of tumor cells in 7/7 cases of adrenal cortical carcinoma, 8/17 cases of cortical adenoma and endothelium in 2/7 cases of adrenal cortical carcinoma and 5/17 cases of cortical adenoma. No immunoreactivity was observed in tumor adjacent adrenal gland. Therefore, IFN-gamma is associated with cell differentiation, the higher the immunoreactivity, the lower the differentiation. The capillary endothelium in adrenal cortical tumor tissue may possess receptor for IFN-gamma or can produce IFN-gamma. PMID- 8732097 TI - [Study on neuropeptide Y-containing innervation of cerebral small arteries and arterioles in spontaneously hypertensive rat]. AB - To comprehend the relationship between the distribution of perivascular neuropeptides and remodified structure of arterioles in hypertension, we applied the technique of sheeting from pia matter of spontaneously hypertensive rats (SHR) to reveal branches at all levels of the vascular network so as to make an overall and stereoscopic observation of the vasculature. In addition, immunohistochemical technique and image analyser were applied to analyse qualitatively and quantitatively neuropeptide Y(NPY)-containing innervation in the vasculature. Findings revealed that the surface density of NPY-containing innervation and integral luminous density of positive reaction particle for anti NPY in middle cerebral artery and its branches -small arteries and arterioles of SHR were significantly increased as compared with findings in Wistar Kyoto. The results reveal that an increase in density of NPY-containing innervation of cerebral small arteries and arterioles intensifies action of nutritive adjustment of vascular smooth muscle and is clearly related with increase of contractive excitability of artery, hypertrophy of artery, smooth muscle cells and the development of hypertension. PMID- 8732098 TI - [Gastrointestinal smooth muscle tumors: a study of DNA content and the association with clinicopathology]. AB - Quantitative analysis of DNA content and ploidy pattern was performed in 84 cases of smooth muscle tumors of the gastrointestinal tract by flow cytometry, and the results were correlated with histopathologic features. The aneuploid rate was found to be 0, 50%, 75%, 92.8% and 100% respectively in the tumors with mitotic figures of 0,1, 2-4, 5-9, or over 10/10HPF (P < 0.005), as to be 3.2%, 60%, 95.8% and 100% in tumors with zero, mild, moderate or severe cellular atypia (P < 0.005). There were no aneuploidy in the 30 cases of leiomyomas, but all 40 cases of leiomyosarcomas were aneuploidy, while in the 14 potentially malignant smooth muscle tumors, there were 4 cases of aneuploidy (P < 0.01). The aneuploid rate was significantly higher in tumors with contiguous organ inveasion or distant metastasis (79.5%), and in tumors of large size (64.5%) as well as in tumors with center necrosis (68.3%). 5 year survival rate in patients with aneuploid tumors (41.6%) was significantly lower than those with diploid tumors (96%) (P < 0.005). The results suggested that DNA content and ploidy pattern be used as an objective parameter to distinguish malignants from benigns and predict the prognosis of patients with gastrointestinal smooth muscle tumors. PMID- 8732099 TI - [Advances in the molecular pathological study of hepatitis viruses and the canceration of liver cancers]. PMID- 8732100 TI - [Relation between loss of heterozygosity at APC and MCC genetic loci and biological behaviour of gastric carcinoma]. AB - In order to detect loss of heterozygosity (LOH) at APC and MCC genetic loci in gastric carcinoma, the authors established a micro-wax-mediated hot start PCR technique. This method allowed a specific gene amplification, and it was useful especially in the amplification of formalin-fixed or stained tissues. In 44 cases of gastric cancer, 29 cases were informative of the APC locus. LOH was found in 8 cases (27.6%): 2 cases in 2 moderately well-differentiated cancer, 2 cases in 13 differentiated cancer, and 4 cases in poorly-differentiated cancer. One of the 3 cases of gastric cancer at early stage also showed LOH. LOH at MCC locus was detected in only 2 of the 25 (8.0%) gastric cancer patients informative. These data suggest that abnormality of APC gene plays a role in the tumorigenesis of gastric cancer and the change may occur at the early stage of tumor development. PMID- 8732102 TI - [The cytochemical observation of inosine effect on glucose metabolism of BGC-823 human gastric carcinoma cell line]. AB - The activity of intracellular lactic acid dehydrogenase (LDH) and succinic acid dehydrogenase (SDH) was studied by cytochemistry in BGC-823 gastric carcinoma cell line treated with inosine. Cytoplasmic granules of LDH were found decreased or completely disappeared, while little change of SDH was observed. The tumor cells were decreased in number and became atrophic, especially when the cells were treated with high concentrations of inosine. The results suggest that inosine can play a role in suppressing the activity of intracellular LDH, thus blocking the major source of energy supply of cancer cells. PMID- 8732101 TI - [Activation of proto-oncogenes induced by MNNG on primary culture of human gastric epithelium and immortalized human gastric epithelial cell line]. AB - Epidemiologic study has shown the association of nitrosamide compounds with the high incidence of stomach cancer in south China. To study the mechanism of gastric carcinogenesis, we have established an immortalized human gastric epithelial cell line GES-1. GES-1 cells and the normal gastric tissues were treated with different concentrations of MNNG for 24 hours. Point mutation at codon 12 of c-Ha-ras gene was found in cells and tissues (43%) as demonstrated by PCR-RFLP. Rearrangement of c-met gene and amplification of c-erbB2 gene were detected by Southern blot assay on the MNNG treated GES-1 cells. The results indicate that MNNG treatment was intimately associated with the activation of certain oncogenes. H-ras and c-met genes, serving as early targets of carcinogens may play important role in the carcinogenesis of human gastric epithelial cells. PMID- 8732103 TI - [Detection of point mutation of p53 gene by silver staining PCR-SSCP in paraffin embedded malignant fibrous histiocytoma]. AB - Silver staining PCR-SSCP method was used to detect point mutation of p53 gene in paraffin-embedded malignant fibrous histiocytoma (MFH) tissues. The abnormal shifting of the single-stranded DNA (ssDNA) was identified in 9 out of 16 cases. The positive figure of SSCP was 1,4,4, 3 in exon 5, 6, 7, 8, respectively. The mutant p53 protein was detected by microwave oven treatment and ABC immunohistochemistry. Positive nuclear staining was observed in 10 cases. The positive coincidence rate was 90.0% between SSCP and p53 protein expression. The mutation of p53 gene was not correlated with the subtypes of MFH. Our results indicate that detection of point mutations with silver staining PCR-SSCP is convenient, rapid and reliable in the screening of point mutation of genes. PMID- 8732104 TI - [Expression of c-myc gene and biosynthesis of biological macromolecules in antisense transfectant HLR60-9]. AB - The recombinant plasmid PGC was constructed for transcription unit of c-myc gene with bi-direction in vitro, to make RNA probes for detection of c-myc mRNA and antisense RNA expression of transfectant HLR60-9, which was obtained from HL60 cells transfected with inducible c-myc antisense RNA expression plasmid. The results from HLR60-9 cells induced by Cd2+ indicated that expression of c-myc antisense RNA increased with Cd2+ concentration and exposure time, while c-myc mRNA expression progressively reduced. Using immunohistochemical technique no c myc P62 protein expression was detected. The incorporation of 3H-TdR,3H-UR,3H-Leu revealed significant suppression of DNA, RNA and protein biosyntheses. It is suggested that the reversion changes previously reported in malignant phenotypes of HLR60-9 cells and the inhibition of macromolecule biosynthesis mentioned above were associated with the blockade of c-myc gene expression by its antisense RNA. PMID- 8732105 TI - [The radiobiologic characteristics of DNA polymerase beta in hepatomas]. AB - DNA polymerase beta activity, its content and gene transcription levels in SMMC LTNM hepatoma were investigated, using 3H-TTP incorporation, immunocytochemistry and cytoplasmic dot hybridization, respectively. The relations between the biological properties of the enzyme and DNA repair synthesis induced by gamma-ray irradiation were also studied. It was found that DNA polymerase beta activity, its content and the amount of its mRNA were much higher in hepatoma than those in normal hepatocytes (P < 0.01). Following whole-body irradiation of the nude mouse bearing SMMC-LTNM with 2 Gy of gamma ray, the polymerase beta activity in hepatoma increased temporarily and the gene transcription of the enzyme seemed to be more active. DNA polymerase beta participated in DNA repair synthesis and this effect was different between hepatoma and hepatocyte because of the biological differences of DNA polymerase beta. The results presented here indicated that DNA polymerase beta could affect radiation damage and radiotherapy of cancer. PMID- 8732106 TI - [Latency of Epstein-Barr virus and its relationship to nasopharyngeal carcinomas]. AB - Non-radioactive in situ hybridization and image analysis were used to study the distribution of Epstein-Barr virus (EBV) DNA and expression of the EBV latent genes, BNLF1, and BamH1 W sequence, in biopsy specimens of nasopharyngeal carcinoma (NPC) and chronic nasopharyngitis. The results showed that the high copy numbers of EBV genome and the expression of BamH1 W sequence, which was related to the transcription of EBV nuclear antigen 1 (EBNA 1) and the initiation of viral replication, occurred almost simultaneously in all poorly- or un differentiated NPC. Furthermore, latent membrane protein (LMP) mRNA was expressed even in the nasopharyngeal epithelium with atypical hyperplasia. These results suggested that enormous replication of EBV episomes occurred with the progression of NPC and might be resulted from the activation of the viral promoter in the poorly- or un-differentiated stage of NPC. The inappropriate expression of LMP might be responsible for the proliferation and de-differentiation of nasopharyngeal epithelium. PMID- 8732107 TI - [Human papillomavirus infection and p53 gene mutation in primary lung cancer]. AB - Surgical samples from 40 cases of primary lung cancer were studied by in situ hybridization and polymerase chain reaction (PCR) to detect HPV-DNA in different types of lung cancer. In addition, PCR-RFLP was used to examine mutation of p53 exon 7. The results showed that HPV-DNA positive rate in lung cancer was 55% (22/40 cases), including SCLC (9/9 cases), squamous cell carcinomas (8/16 cases), and adenocarcinomas (5/12 cases). Amplification of p53 exon 7 was seen in 5 of 22 HPV-DNA positive cases. RFLP analysis showed that p53 exon 7 mutation was present in two cases with gene amplification. In HPV-DNA negatives there was only one case with exon 7 amplification but no mutation. SCLC and squamous carcinoma had higher HPV infection rate than other types of lung cancer. The results suggest some relation between HPV infection and p53 gene mutation. PMID- 8732108 TI - [ACNU and methyl-CCNU in combination chemotherapy for advanced gastric cancer: a randomized comparative study. Shanghai ACNU Collaborative Study Group]. AB - The study was conducted to compare the efficacy between a combined chemotherapy of ACNU (Nimustine hydrochloride, brand name Nidran), a new nitrosourea agent, and a combined chemotherapy of methyl-CCNU for advanced gastric cancer. One hundred and three cases of advanced gastric cancer were randomly allocated into Group A (Me-CCNU, 5-Fu and ADM) and Group B (ACNU, 5-Fu and ADM). The results showed that in Group A there was no CR or PR, while in Group B, although no CR, there were 8 PR (8/64 cases), denoting the response rate of 0% in Group A and 17.4% in Group B. The median survival period in Group A was 108 days and in Group B 112 days. Both groups well tolerated the treatment and there were no serious adverse effects. Quality of life (QOL) was better in Group B than in Group A. It is concluded that ACNU combination is superior to Me-CCNU combination for advanced gastric cancer. PMID- 8732109 TI - [Changes in hematoporphyrin in patients with gastric and hepatic carcinoma and their clinical significance]. AB - The fluorescence spectroscopy of porphyrin molecule in blood was determined with photoluminescence fluorescence spectroscope in 139 patients with gastric carcinoma, 76 with hepatic carcinoma, 110 with gastric ulcer and chronic gastritis, 168 with liver cirrhosis and 33 normals as controls. The results showed that the peaks of Zinc porphyrin and protoporphyrin in patients with cancer were significantly higher than those in patients with benign disease, the peak of protoporphyrin being two to three times higher in the former groups of patients than that in the latter (P < 0.01). Protoporphyrin could be used as a marker to screen and diagnose gastric and hepatic carcinoma. PMID- 8732110 TI - [Supra-aortic arch esophagogastrostomy through esophageal bed in treating patients with middle and lower esophageal carcinoma]. AB - A new surgical procedure of esophagogastrostomy was carried out in 162 cases with middle or lower thoracic esophageal carcinoma in our department from January, 1992 to December, 1994. Esophagogastrostomy was performed at supra-aortic arch site by pulling the stomach along the esophageal bed instead of putting it before aortic arch in the thoracic cavity. The procedure may keep the normal physiological function of the organs in the chest. It may help fully distend the left lung so as to reduce impairement of pulmonary function because the thoracic stomach was kept within the mediastinum. No resection mortality or anastomotic leakage occurred in the series. Since the esophagus and the stomach were anastomosed in a linear fashion without angulation, the evenly distributed intracavitary pressure may help prevent anastomotic leakage from occurrence. It is also easy to be dilated if anastomotic stricture occurs. In addition, the new procedure may be helpful in promoting gastric emptying, preventing gastric reflux, depressing and stopping the blood oozing in the mediastinum and air leakage from the ruptured lungs, as well as in distinguishing pleural effusion from gastric retention. Compared with other procedures, it is more difficult to perform the anastomosis. Once anstomotic leakage occurs, drainage is not easy. PMID- 8732111 TI - [Solitary plasmacytomas of bone and extramedullary plasmacytomas]. AB - Among plasma cell disorders, solitary plasmacytoma (solitary plasmacytoma of bone, SPB and extramedullary plasmacytoma, EMP) is rare as compared with multiple myeloma (MM). Furthermore, the relationship between solitary plasmacytoma and MM remains unclear. Between 1960 and 1994, 24 patients with SPB and 20 with EMP were treated. The criteria for diagnosis were: (1) no evidence of other lesions based on clinical and radiologic examinations; (2) biopsy evidence of a plasma cell neoplasm; (3) bone marrow biopsy specimen with negative findings (less than 10% plasma cells); (4) no anemia, hypercalcemia or renal involvement. The average follow-up period was 112 months (from 6 to 360 months). Fifty-four percent of patients with SBP and 40% of patients with EMP developed MM, however, there was no significant statistical difference between SPB and EMP (P > 0.05). We suggested that solitary plasmacytomas be classified as two types, latent and aggressive. The former was histologically well-differentiated plasmacytoma. The latter was poorly differentiated tumors which easily progress to MM. The treatment of choice is wide excision or thorough curettage, by cryogenic necrosis with liquid nitrogen or cautery of the bony wall with phenol and the cavity filled with bone grafts or cementation. All patients with apparently isolated plasmacytoma should receive local radiotherapy after operation. Adjuvant chemotherapy should be given if the tumour turns out to be poorly differentiated, in order to delay their progression to MM. PMID- 8732112 TI - [Clinical evaluation of cervical esophageal reconstruction after resection of thoracic esophageal carcinoma]. AB - From Apr. 1979 to Dec. 1994, a total of 3,714 patients with thoracic esophageal carcinoma were surgically treated. As a standard operative procedure for thoracic esophageal cancer, cervical esophagogastrostomy was performed. While this operative procedure was rarely used (accounting for only 8.8% the treated cases) in early years, it has been recently carried out in the overwhelming majority of our patients (94.1%). This technic offers the advantages of resecting the lesions radically, dissecting the regional lymph nodes in both the neck and the mediastinum, reducing the incidence of postoperative complications and improving the quality of life after operation. In this paper the operative indication, approach and the surgical efficacy are discussed. PMID- 8732113 TI - [Diagnosis and treatment of pleural mesothelioma: a report of 40 cases]. AB - Forty patients with pleural mesothelioma confirmed pathologically including 2 benign and 38 malignant lesions are reported. In the 38 malignant mesotheliomas, 12 were histopathologically defined while the rest were unclassified. In this series, 33 surgical resections and 3 thoracotomy biopsies were carried out, whereas 4 patients were not operated. In 36 thoracotomies, there was one death with an operative mortality of 2.8% (1/36). Of 33 surgical resections, 10 (31%) received combined therapy. Of the 3 thoracotomy biopsy cases, postoperative irradiation was given to one, irradiation plus chemotherapy to the second and no treatment to the third. In 4 who were not operated, one died without treatment, two accepted chemotherapy and the fourth was treated with irradiation plus chemotherapy. Only two (5%) of these 40 patients had history of exposure to asbestos. The pre- and intra-operative diagnostic conformity rate was 50%. Ten patients were lost from follow-up. The longest survival was twelve years and one month. The authors sum-up their 31-year (1963-1994) experience of this disease and discuss the occupational factors in etiology, clinical diagnosis, treatment options and the significance of pathological types on prognosis of pleural mesothelioma. PMID- 8732114 TI - [Intracavitary microwave hyperthermia combined with external irradiation in the treatment of esophageal cancer]. AB - A randomized trial of intracavitary microwave hyperthermia combined with external irradiation (R + H) versus radiation (R) alone in the treatment of esophageal cancer was performed from Feb. 1986 to Feb. 1988. In the R group, radiation was given by 8 MV X-ray with 2 Gy/fraction, 5 fractions per week with a total dose of 60 Gy/6 weeks. In the R + H group, the radiation was given as R group but with a total dose of 40 Gy/4 weeks. Intracavitary 915 MHz microwave hyperthermia was given with a nominal temperature of 43.5 degrees C at the margin of the tumor surface, 45 minutes/session, 1-2 sessions/week for 4-8 sessions. The 1-, 3-, and 5-year survival rates in R + H group were 81.2% (48/59 cases), 42.4% (25/59) and 23.7% (14/59), while in the R group 59.0% (39/66 cases), 24.2% (16/66) and 16.7% (11/66) respectively. The differences in 1- and 3-year survival rates were statistically significant (P < 0.05) between the 2 groups. Using the thermal dose T90 analysis, after the cases with T90 < 43 degrees C (insufficient thermal dose) were eliminated, 52 cases with T90 equal to or higher than 43 degrees C had 1, 3, and 5 year survival rates of 84.6%, 44.2% and 26.9%, respectively. Statistically significant differences in the 2 groups were also limited only to 1- and 3-year survivals. Higher 5-year survivals is anticipated if more cases are studied. PMID- 8732115 TI - [Surgical treatment of primary mediastinal tumor: a report of 90 cases]. AB - In the past 22 years 90 cases of pathologically confirmed primary mediastinal tumors (PMT) were surgically treated. Of these 90 cases, there were 38 thymomas (42.3%), 19 teratoblastomas (21.2%), 9 neurofibromas (10.0%) and 24 others (26.5%). Radical excision was performed in 70 (77.8%), palliative excision in 6 (6.7%), exploratory thoracotomy in 12 (13.3%). Postoperative death occurred in 2 (2.2%). No relapse was reported in patients who had received radical excision during the follow-up period of 3 months to 13 years. Besides careful history taking and physical examination, X-ray examinations of the chest, particularly CT scan were valuable in the diagnosis and differential diagnosis of PMT. Difficulties in diagnosis were due to unusual pathological changes and clinical manifestations. Due precautionary measures should be taken to avoid injury to the heart, major blood vessels and the spinal cord. Cord injury was the consequence of direct operative trauma, compression due to intraspinal hemorrhage, and/or operative interruption of blood supply. PMID- 8732116 TI - [Expression of p53 in human esophageal carcinoma and pre-malignant lesions in the esophagus]. AB - The levels of p53 protein were detected by immunocytochemistry in 197 endoscopic biopsy specimens of esophagus. Nuclear p53 protein was present in 2.9% of normal mucosa, 6.7% of inflammatory mucosa, 38.8% of mild dysplasia, 52.0% of moderate or severe dysplasia, 61.1% of carcinomas in situ, 62.5% of invasive carcinomas. Moreover, we examined 14 cases of esophageal carcinoma with extensive dysplasia near the carcinoma and found high levels of p53 protein in both dysplasias and carcinomas in 8 cases. In one case, carcinoma presented high level of p53 protein while the adjacent dysplasia did not. In another case, the result was just the opposite. Negativity for p53 immunostaining was found in 4 cases. These data indicated that p53 protein accumulation occurred before tumor invasion in the multistage esophageal carcinogenesis. The timing and the frequency of p53 protein accumulation made p53 gene an attractive marker for the early diagnosis an the evaluation of chemopreventive agents. PMID- 8732117 TI - [Study on ultrastructure morphology of fine needle aspiration of mediastinal and lung lesions]. AB - Specimens of Mediastinal lesions and lung masses collected by fine needle aspiration (FNA) was examined with both light and transmission electron microscopy. Sixty eight specimens (9 mediastinal and 40 lung masses, 4 metastatic chest masses and 15 metastatic lymph nodes from lung cancer) from 104 cases (60.5%) were satisfactory for evaluation. With light microscopy, the nature of the lesions was assessed and the ultrastructural features of the lesions were evaluated with transmission electron microscopy. Diagnosis of histologic type of the lesions by light microscopy was consistent with that by transmission electron microscopy at a conformity rate of 50.4%. The results indicate that ultrastructural features are valuable in ascertaining histologic type of the lesions. PMID- 8732118 TI - [Prognostic factors affecting the results of surgical treatment of cervical cancer]. AB - From October, 1963 to December, 1992, 1,213 cases with cervical cancer were treated surgically in our hospital, among whom 922 cases, including 305 in stage 0, 123 in stage IA, 212 in stage IB, 265 in stage IIA, 17 in stage IIB, were operated before February, 1990. Among the 617 evaluable cases, excluding those in stage ), the five-year survival rates were 95.1% in stage IA, 91.0% in stage IB, 83.1% in stage IIA and 59.0% in stage IIB, respectively. The results showed that cervical tumour greater than 4 cm in diameter, invasion in muscular layers, lower degree of differentiation and pelvic lymph nodes metastasis would lead to worse therapeutic effects. The method of pelvic lymphadenectomy, pathological types, and ages of the patients, however, did not at all correlate with the survival rates of the patients. For those who have above-mentioned risk factors active adjuvant treatments are indicated. PMID- 8732119 TI - [Thoraco-cranial operation for treatment of lung cancer with brain metastasis]. AB - Ten cases of lung cancer with brain metastasis who received simultaneous surgery for the primary lung tumor and for the metastatic intracranial tumor between 1990 and 1994 are reported. There was no death due to operation, nor were serious complications. With follow up until Feb. 1995, four patients died and its mean survival period was 8.25 months. Six patients are still alive with a mean survival period of 16 months, the longest one being nearly 3 years. The authors hold that, if the primary tumor is located at the periphery of the lung, the brain metastasis is solitary and patient's general condition is good, simultaneous thoraco-cranial operation is indicated and feasible. Postoperative adjuvant chemotherapy is needed to achieve better treatment results. PMID- 8732121 TI - [Prospective study on intelligence quotient of 28 hepatic lenticular degeneration patients with treatment combining traditional Chinese and Western medicine]. AB - Intelligence quotient (IQ) of 28 hepatic lenticular degeneration (HLD) patients was analysed with Wechsler Adult Intelligence Scale-revised by China (WAIS-RC) before and after two months treatment by the HLD-Relief decoction supplemented with copper-clearing drug, and the result was compared with that of 30 healthy persons for control. The results showed that the Vocable IQ (VIQ), Performance IQ(PIQ) and Full IQ(FIQ) of HLD patients were markedly lower than that of control (P < 0.005), especially in visual space disorder, which indicated that the copper ion depositing in patient's brain not only damaged neuronic motorial functions but also produced a marked effect on the patient's intelligence. After treatment, the total intelligential level including VIQ, PIQ and FIQ has been raised, particularly in PIQ in which all scores were obviously increased (P < 0.001). It was also showed that the damage of intelligence in HLD patient became more serious as the course of disease was prolonged and the early treatment facilitated the IQ level to approach the normal criterion. It indicated that the treatment with combined therapy is effective to the intelligence and extrapyramidal symptoms in HLD patients. PMID- 8732120 TI - [Combination treatment of late nasopharyngeal carcinoma: an analysis of short term therapeutic results in 200 cases]. AB - Patients with late nasopharyngeal carcinoma (NPC) were divided into 4 groups, 50 each, treated with different protocols. Group A: radiotherapy plus systemic interlenkin 2 (IL-2), Group B: chemotherapy plus radiotherapy, Group C: chemotherapy plus radiation and IL-2, Group D: radiotherapy alone. Changes in the primary tumor and neck lymph nodes were monitored at 0, 3 and 6 months after treatment. In group A, the primary tumor regression rates (TRR) were 88%, 90% and 92%, and the lymph nodes regression rates (LNR) were 82%, 86% and 88%, respectively. In group B, the TRR were 86%, 88% and 90%, and the LNR were 80%, 84% and 86%, respectively. In group C, the TRR were 88%, 90% and 92%, and the LNR were 82%, 86% and 88%, respectively. In group D, the TRR were 78% 80% and 82%, and the LNR were 76%, 78% and 80%, respectively. The 1-year survival rates in these 4 groups of patients were 92%, 90%, 94% and 82%, respectively. Thus, the therapeutic results are significantly better in the combination treatment groups than those in the group receiving radiotherapy alone. In addition, IL-2 was effective to significantly reduce the occurrence of systemic side effects of radiotherapy and radiotherapy plus chemotherapy. PMID- 8732122 TI - [Clinical and experimental study on sustained release tablet of Tripterygium wilfordii in treating rheumatoid arthritis]. AB - Adopt to the prospective, multi-center, random, single-blind, equal rank-control methods, 226 patients of rheumatoid arthritis diagnosed according to the ARA criteria, were divided into 2 groups. One hundred and fourteen patients of test group were treated with sustained release tablets of Tripterygium wilfordii (TW SR) orally, 2 tablets, twice a day for 4 weeks, 112 patients of control group received tablets of Tripterygium wilfordii (TW) orally, 2 tablets 3 times per day for 4 weeks. Results showed that the total effective rate of the two groups were 92.11% and 90.65%, respectively (P > 0.05). The adverse reaction rate of TW-SR group was 20.18%, which was lowered than that of TW group (70.54%, P < 0.01). Results of pre-clinical pharmacologic experimental study showed that the TW-SR has obvious anti-inflammatory, analgesia and immunosuppress'ive action as the TW has, while its toxicity was less than the latter significantly. PMID- 8732123 TI - [Clinical and experimental study of RA mixture in treatment of rheumatoid arthritis]. AB - The RA mixture is composed of Tripterygium Wilfordii (TW) and other Chinese medicinal herbs with effect of expelling Wind, activiting blood circulation, invigorating the Kidney and Qi. The authors treated rheumatoid arthritis (RA) patients with RA mixture and compared it with D-penicillamine as control. The two group's therapeutical effect is similar (P > 0.05), but the side effect occuring ratio in treatment group was obviously lower than that of control group (P < 0.01). After treatment with RA mixture, the patient's human lymphocytic antigen degenerative reaction (HLA-DR+) cell, CD4/CD8 ratio reduced and auto-mixed lymphocytic reaction (AMLR) level enhanced (P < 0.05). The results of animal experiment showed that swelling of RA model mouse's joint could be reduced by both RA mixture and TW. Comparing with TW, RA mixture had stronger effect in controlling the inflammation of synovial cell and fibroid degeneration of fibrocytes. At the same time, RA mixture had stronger effect in protecting the immune organ from atrophy and protecting functions of cellular immunity. All above suggest that RA mixture reduce the syndroms of RA by improving distribution of T lymphocyte subsets and the functions of cellular immunity. The other Chinese medicinal herbs in the RA mixture could enhance TW's therapeutical effect and reduce it's side effect. PMID- 8732124 TI - [Clinical study on effect of jinshisan on dissolution of extra-/ intrahepatic biliary stones]. AB - In order to evaluate the role and mechanism of Jinshisan (JSS) in bile stone dissolution, 164 cases of extra-/ intrahepatic biliary stone patients were treated by a collaborated group, the therapeutic effect was observed by B ultrasonography before and after treatment. Results showed: (1) Among the 164 cases, 26 cases (15.85%) were cured clinically; 73 (44.51%) were effective and 65 (39.63%) ineffective. (2) After treatment, in 84 patients of right hepatolith, the stone were diappeared in 10 cases, the size or number of stone decreased in 44 casess. In 34 patients with left hepatolith, the figure were 5 and 13 respectively. In 10 cases of choledocholith they were 10 and 10 respectively. While in 21 cases of multiple gallstone, only one case with stones of both sides disappeared, except 3 ineffective cases, others with stone decreased in size or number or disappeared unilaterally. (3) Gallston diappeared in 7 of 10 patients with size of stone 6 months. (5) Analysis of bile showed that JSS could increase the amount of bile acid, decrease the bilirubin, mucous and bicarbonate radical in bile. PMID- 8732125 TI - [Effect of integrated Chinese medicine on early stage of acute cerebral hemorrhage]. AB - The effect of integrated Chinese medicine (ICM) on the convalescent of acute cerebral hemorrhage patients were observed. The results showd that the effect of patients treated with ICM was better than that treated with Western medicine alone on the aspects of absorption of brain hemetoma, elimination of encephaledema and recovery of neural function (P < 0.05, 0.01). It suggested that early application of integrated Chinese medicine is very helpful in rehabilitation of cerebral hemorrhage patients. PMID- 8732126 TI - [Clinical study on the effect of shuxuening tablet in treatment of coronary heart disease]. AB - Shuxuening was made of extract of Folium Ginkgo, the Shuxuening tablet No. 2 was a pure extract and No. 1 was a crude extract. Forty-six coronary heart disease patients were divided randomly into two groups according to the ratio of 2:1, Shuxuening tablet No. 1 and No. 2 were given to the two groups respectively. Results showed that both of them could alleviate the symptom of angina, the total effective rate of 2 groups were 60.00% and 83.87%, there was no significant difference between the two groups. But for the angina of middle and severe degree, the total effective rate of No. 2 was 83.33%, while that of No. 1 was 33.33%, the difference was significant (P < 0.01). They could also improve the abnormal electrocardiogram, the total effective rate of two groups were 60.00% and 74.19%, no significant difference between them was found. The two drugs could decrease the blood lipid as well. PMID- 8732127 TI - [Clinical study on treatment of chemo- or radiotherapy induced leukopenia with fuzheng compound]. AB - The therapeutic effect of Fuzhen Compound, a Chinese herbal medicine, on leukocyte count in 59 patients with cancer of advanced stage after chemo-or radiotherapy were observed. Among 41 cases with leukopenia before treatment, leukocyte of 37 cases were normalized after treatment, the count pre-and post treatment were (2.8 +/- 0.6) x 10(9)/L and (5.9 +/- 1.4) x 10(9)/L respectively (P < 0.01). In 18 patients with normal leukocyte count before treatment, the counts of 17 cases were still kept in normal after treatment. The time for leukcoyte recovering was 8.0 days in average, while that of 23 patients did not take Fuzhen Compound was 20.6 +/- 1.8 days. The results indicated that the Fuzhen Compound could alleviate the inhibition of chemo-or radiotherapy on hemopoietic function of bone marrow and has leukocytogenic effect. PMID- 8732128 TI - [Study on relationship of lipid peroxide in coronary heart disease with and without diabetes]. AB - Activity of erythrocyte superoxide dismutase (RBC-SOD), levels of serum malondialdehyde (MDA), plasma 6-keto-prostaglandin F 1 alpha (6-keto-PGF 1 alpha) and thromboxane B2(TXB2) were measured in 30 healthy subjects and 57 coronary heart desease (CHD) patients inoluding 21 cases complicated with diabetes and 36 without. Their characteristics of Syndrome Differentiation and typing were observed. The results showed that the activity of RBC-SOD, 6-keto-PGF 1 alpha in CHD patients were significantly lower than those in the healthy subjects, but the levels of TXB2, MDA were significantly higher. The levels of TXB2 in CHD patients with diabetes were significantly higher than tose without, but the activity of RBC-SOD were significantly lower. There were remarkably positive correlations between the levels of serum MDA and that of blood sugar, TXB2, TG and BMI in CHD patients with diabetes. There were remarkably negative correlations between the levels of serum MDA and plasma 6-keto-PGF 1 alpha. The results suggested that the metabolic abnormality of lipoperoxides was more serious in CHD patients with diabetes than without. The main Syndrome of CHD with diabetes was Qi-Yin Deficiency with Blood Stasis, while that of without diabetes was Qi Deficiency with Blood Stasis. PMID- 8732129 TI - [Effect of kanggusong in prevention and treatment of retinoic acid induced osteoporosis in rats]. AB - Retinoic acid 70 mg/kg.d was given by gastrogavage to Wistar rat for 14 days to induce osteoporosis. Kanggusong (KGS), a mixture of extracts from 8 traditional Chinese drugs, was given to 3 test groups of rats simultaneously in various dosage. Results showed that the KGS displayed obvious action in preventing osteoporosis, the trabecular loss of tibiae and bone loss of compact bone were lowered markedly in KGS groups with high (3.0 g/kg.d) or middle (1.0 g/kg.d) dosage in comparing with control model group, the trabecular area percentage and compact bone area percentage were increased significantly (P < 0.05) which approached to the level of normal control group. KGS could also improve the pathological changes in microstructure of bone, increase the thickness of trabecula and cortex (P < 0.05), reduce the trabecular gap and bone marrow cavity (P < 0.05). The mechanism of KGS might be relevant with its action of suppressing the osteoclast activity and activating osteoblast, resulting a positive balance of bone metabolism, increasing the blood concentration of calcium and estrogen as well as its antagonistic action against the injury of sex glands by retinoic acid. PMID- 8732130 TI - [Experimental study of R plasmid eliminating action of Coptis chinensis on E. coli]. AB - The R plasmid curing experiment was performed in vitro with E. coli strain E. 102 bearing R plasmid as target bacteria and Coptis chinensis as elimination agent. The influence of different time on R plasmid elimination was also observed. Results showed that when the acting time was 24 hours, the cure rate of R plasmid was 2.42% and when the acting time increased to 48 hours, the cure rate elevated to 22.57%. The Missing patterns of R plasmid might be occurred in disappearence of either multiple or single resistance. PMID- 8732131 TI - [On traditional Chinese medicine-mediated biotherapy of leukemia]. PMID- 8732132 TI - [Survey on experimental studies on traditional Chinese medicine treatment of chronic renal failure]. PMID- 8732133 TI - [Progress in the treatment of coronary heart disease with antioxidant of integrated traditional Chinese medicine-Western medicine]. PMID- 8732135 TI - [Clinical study on effect of combined treatment of fuchunpian with radiotherapy on nasopharyngeal carcinoma]. AB - The study based on the clinical prospective trial of 60 cases of nasopharyngeal carcinoma treated with combined treatment of radiotherapy and Fuchunpian in our hospital from September, 1988 to January, 1990. Results of clinical data of 5 years follow-up showed that Fuchunpian not only couldn't enhance the radiosensitivity of patients, but also increase the blood metastatic potency, the metastatic rate of patients treated with Fuchunpian (36.7%) was 2.67 times higher than that without it (10.0%). Therefore, it was considered that use Fuchunpian as a routine either administrated alone or combined with radiotherapy in treatment of nasopharyngeal carcinoma is not suitable. PMID- 8732134 TI - [Nuclear cardiology study on effective ingredients of Astragalus membranaceus in treating heart failure]. AB - The experimental study has testified that among the various effective constituents gained from Astragalus membranaceus (AM) is the main component. Nineteen patients with heart congestive failure were treated with effective ingredient of AM, the astragaloside IV (XGA) injection. RESULTS: After 2 weeks of treatment the symptoms of chest distress, dispnea in 15 patients was alleviated, their capability of exercise reinforced. Radionuclide ventriculography showed that left ventricular modelling improved, left ventricular end-diastolic volume diminished by 11.74 +/- 18.39 ml, left ventricular end-systolic volume by 9.35 +/ 18.01 ml, with statistical significance. HR slowed from 88.21 +/- 17.19 to 64.55 +/- 13.06 beats/min, P < 0.05; PER increased from 1.80 +/- 0.86 to 1.95 +/- 0.85 u/second, P < 0.05. Left ventricular EF, PFR increased also at some extent without statistical significance. CONCLUSION: Effective ingredient of AM, XGA injection is efficient positive inotropic drug, and could improve the left ventricular modelling and ejection function in patients with congestive heart failure after continuous administration of XGA injection for two weeks. PMID- 8732136 TI - [Clinical observation on child aplastic anemia treated with integrated Chinese and Western medicine]. AB - Forty-three cases of aplastic anemia were treated with fetal blood transfusion, Chinese medicinal herbs and Vit. C. The results showed that the effective rate of the treated group was 79.1%, among them, the chronic aplastic anemia (CAA) was 88.9%, acute aplastic anemia (AAA) was 62.5%, both rates were higher than that of the control group (with western medicine alone, n = 46). The difference of treatment results between two groups was highly significant. The mechanism was that the therapy could rebuild hematopoietic function, modulate immune function and improve microcosmic environment in bone marrow so that it promoted in all aspect the recovery of hematopoietic function in organism. PMID- 8732137 TI - [Study on ultrastructure of platelets in migraine patients with blood stasis type]. AB - To study the relationship between ultrastructure of platelets and migraine patients with blood stasis type (MBS). The morphologic appearance and aggregation of platelets in MBS patients were examined before and after treatment with Tongbike oral liquor (20 cases in treated group and.10 in control). The results showed that the MBS patients' platelets appeared irregular, easy to aggregate and adhere to red cells, which was more severe when ADP was added, but improved after treated with Chinese medicinal herbs. According to the results it was assumed that platelets from MBS patients were in an abnormal activated state, the abnormal morphology and functional disturbance might be the pathologic basis of MBS. Tongbike oral liquor could markedly improve the morphologic and functional state of platelets, which was relevant to the inhibition of platelet aggregation and the inducing of platelet depolymerization. PMID- 8732138 TI - [Study on mitochondrial ultrastructure, trace elements and correlative factors of gastric mucosa in patients with spleen deficiency syndrome]. AB - Eighty-eight gastropathic patients with Spleen deficiency syndrome by using transmission electron microscope (TEM), X-ray energy disperse analysis system (EDAX), histochemical staining and radioimmuno methods were examined. The authors found that the gastric mucosa cAMP, SOD level, the quantity of mitochondria and its crista, the ratio of diameter between ventricle and cavity of mitochondria and the content of Zn, Cu of mitochondria were reduced in the trend of healthy control group, Spleen Qi deficiency group, Spleen deficiency with Qi stagnation group; chronic superficial gastritis group, chronic atrophic gastritis group, gastric cancer group: complete small intestinal metaplasia group, incomplete small intestinal metaplasia group, complete colonic intestinal metaplasia group, incomplete colonic intestinal metaplasia group (P < 0.05-0.001). While the degeneration rate of mitochondria, the Cu/Zn ratio of mitochondria, the metaplasia rate of gastric, the rate of incomplete colonic intestinal metaplasia and the content of serum LPO were increased in the above turn. It is suggested that the comprehensive effect of the degeneration of mitochondria and the quantitative changes of its correlative factors is the physiopathologic base for inducing Spleen deficiency disease, gastric mucosa metaplasia and canceration. Much attention must be paid in clinic to the cancerization trend of gastric disease with Spleen deficiency syndrome. PMID- 8732139 TI - [Syndrome differentiation, gastro-intestinal function and electro-gastrography of chronic obstructive pulmonary disease and pulmonary heart disease]. PMID- 8732140 TI - [Effect of yougui yin on the content of monoaminic transmitters and body weight, food and fluid intake in corticosterone-rats]. AB - Great attention has been paid to the relationship between the central monoaminic transmitters and the hypothalamus-pituitary-adrenal (HPA) axis recently. This paper reported the content of hypothalamic monoaminic transmitters of rats, which were injected with corticosterone 10 mg/kg subcutaneously for 14 days. The body weight as well as volume of food and fluid intake were measured in the same period. Effect of Yougui Yin (YGY, administrated by gastrogavage) on he cortico inhibited rats was also observed. Results showed that the contents of hypothalamic monoaminic transmitters such as norepinephrine, Dopamine, 3,4 Dihydroxyphenylalanine, serotonin, and 5-hydroxyindole acetic acid of the rats increased obviously, while the body weight, volume of food and fluid intake decreased significantly day by day. The YGY could improve the above-mentioned indexes significantly. It suggested that: (1) The synthesis and metabolism of the central monoaminic transmitters of rats activated when the HPA axis was inhibited by exogenous glucocorticoid, their body weight lowered, and the volume of food and fluid intake decreased. (2) YGY could improve the inhibited HPA axis, and inhibit the activated central monoaminic transmitters as well as protect the normal regulating function of hypothalamus on food and fluid intake. PMID- 8732142 TI - [Experimental study on the effect of qing-wen granule on epidemic hemorrhagic fever virus]. PMID- 8732141 TI - [Effect of shengmai san on diaphragmatic function in rabbits]. AB - After stimulation of the bilateral phrenic nerves of the rabbits, the effect of Shengmai San (SMS) on their normal function and the diaphragmatic fatigue was observed with the measuring of diaphragm evoked potential (DEP), and the analysis of the power spectrum of diaphragmatic electromyogram (EMGdi) at spontaneous breathing. The results were as follows: (1) After the administration of 2 ml/kg of SMS, amplitude of DEP, central frequency, and the high/low frequency ratio were markedly increased. (2) Injecting SMS in 2 ml/kg in 30 minutes before injury could protect significantly against diaphragmatic fatigue following electrical stimulation of the phrenic nerves. (3) Injecting SMS in 2 ml/kg after fatigue could enhance the recovery of the diaphragmatic fatigue. PMID- 8732143 TI - [Analysis of the pulse harmonic frequency spectrum. A new method of pulse study of traditional Chinese medicine]. PMID- 8732144 TI - [Progress in studies of chronic exhaustion syndrome and its traditional Chinese medicine therapy]. PMID- 8732145 TI - [Analysis of therapeutical effectiveness in 35 cases of acute leukemia with therapy of Chinese integrated medicine]. AB - Thirty-five cases of acute leukemia were treated with methods of Chinese integrated medicine (CIM), i.e. in the CIM group, in which the complete remission rate, partial remission rate and total remission rate were 68.5%, 20.0% and 88.5%, respectively; while those in the random control group were 42.7%,20% and 62.7%, respectively. The comparison between the two groups showed a significant difference(P < 0.05). The patients' survival period in CIM group of 1,3,5 and above 5 years was shown in 20, 12, 7 and 5 cases, but in the control group, 9,5,0 and 0 cases, respectively. The comparison between the two groups also showed an obvious difference (P < 0.01), which demonstrates that the therapeutic effect was better in the CIM group and the patients' survival period was longer in the CIM group, too. PMID- 8732146 TI - [Clinical observation on treatment of 40 cases of apoplexy hemiplegia complicated shoulder-hand syndrome with electro-acupuncture]. AB - The authors treated 40 cases of shoulder-hand syndrome of apoplexy hemiplegia with electro-acupuncture (EA) and filiform needle acupuncture (FNA) respectively. The results showed that EA had better results in treating hand back swelling, hand skin temperature elevating and the bending finger caused pain than that with FNA (P < 0.05). The finger joint and shoulder joint improvement (the functional scoring increased for 3 points or more) in EA was also better than that of FNA (P < 0.05). The total marked effective rate was higher in EA group (75%) than that in FNA (50%), P < 0.05). It suggested that EA produced rhythmic muscle contraction which had a "shoulder-hand pump" like action, and is significant in eliminating hand back swelling and preventing atrophy of hand muscles. PMID- 8732147 TI - [Clinical observation on treatment of 45 angina pectoris patients of coronary heart disease with taponin]. AB - Forty-five patients suffering from angina pectoris of coronary heart disease (CHD) were successfully treated with Taponin. The effect of the treatment was compared with that of nifedipine. The clinical practice showed that the group treated with Taponin yielded better results. After medication, the patients were markedly relieved from angina pectoris of CHD. Meanwhile the improvement of electrocardiogram (EKG) and impedance cardiogram were also observed. The effective rate of EKG improvement was 83.7%. The treatment of angina pectoris of CHD with Taponin was significantly more effective in comparing with control group (P < 0.05). PMID- 8732148 TI - [Effect of age on hemostasis and plasma level of TXB2 and 6-keto-PGF1 alpha]. AB - Hemostasis (HS) in 134 healthy subjects of over 20 years old was investigated. The cases with HS symptom were 45.5%, which increased with age. TXB2, 6-keto-PGF1 alpha and T/K ratio were measured by radioimmuno assay (RIA). RESULTS: elevation of TXB2 was more significant in the middle age and old age than in the young group (P < 0.01). But the level of 6-keto-PGF1 alpha in various age group didn't changed significantly; while the ratio between TXB2 and 6-keto-PGF1 alpha was more significant in the aged than in the young person (P < 0.01). The results revealed that there was hypercoagulable tendency with the increase of age, and it was correlated with TXB2 and 6-keto-PGF1 alpha. It is significant in theory and practice to prevent and cure the cardiovascular and cerebrovascular disease as well as HS with the traditional Chinese medicine. PMID- 8732149 TI - [Correlation between kidney deficiency syndrome and bone density]. AB - In order to study the relationship between Kidney Deficiency Syndrome (KDS)and bone density (BD), the BD of 2068 subjects (40-69 years old)was measured with SPA IIC type monophoton BD scanning device made in China. The subjects including 1144 cases of KDS (320 males and 824 females); 608 cases without KDS (306 males and 302 females); 164 cases of Lung Deficiency (80 males and 84 females); 152 cases of Spleen Deficiency (76 males and 76 females). Through comparison with the same sex and age group, it was found that BD in KDS patients was significantly lower than that in the normal subjects and those without KDS, also lower than that in the Lung and Spleen Deficiency patients. It indicated that KDS had the corresponding BD changes, which confirmed the TCM classical theory "The Kidney governs the bone". Based on the measuring data, the authors took the lower limit of BD normal value minus two folds of SD as the critical value for KDS diagnosis in 40-49 years age group, i. e., male BD < 0.6 g/cm2, female BD < 0.5 g/cm2. The clinical feedback showed that the male and female related coincident rates were 76% and 80% respectively. Therefore, the critical value could be taken as an objective index for KDS diagnosis after excluding other affecting factors. PMID- 8732150 TI - [Study of relationship between level of serum selenium and syndrome differentiation and typing of traditional Chinese medicine in lung cancer]. AB - The relationship between the Syndrome differentiation and typing of TCM and the level of serum Se in the patients with the primary lung cancer was studied. The results revealed that the changed level of serum Se reflected the wax and wane of the pathogenic factors and body resistance in the patients with lung cancer. The level of serum Se gradually decreased as a result of the reducing body resistance and strengthening pathogenic factors, which suggested that it facilitated the evaluation of the patients' condition, recovery and prognosis of lung cancer. Due to the level of serum Se was lowered with the using of some chemotherapeutic agents, and through the treatment of lung cancer with the Syndrome Differentiation of TCM, it could supplement chemotherapy and maintain the serum Se level. It suggested that some Chinese medicinal herbs rich in Se should be selected to treat lung cancer so as to improve the therapeutic effect. PMID- 8732151 TI - [Effect of bushenhuoxue tablet on serum lipid peroxide, blood lipid and blood sugar in type II diabetics]. AB - Sixty-eight patients suffering from diabetes mellitus (DM) type II with Kidney Deficiency and blood stasis were enrolled and divided randomly into treatment group (34 cases) and placebo group (34 healthy subjects). The amount of serum lipid peroxide (LPO), blood lipid and blood sugar was determined. The results showed that the serum LPO in treatment group was higher than that in placebo group. The serum LPO increased significantly in DM patients with vascular disease or with uncontrolled blood sugar. The serum LPO was positively correlated with triglyceride (TG). After using Bushenhuoxue Tablet (BSHX) serum LPO lowered, blood sugar decreased and high density lipoprotein-cholesterol (HDL-C)increased in treatment group, but in placebo group these three parameters were not changed significantly. These results indicated that LPO reaction in type II DM patients increased. The higher the blood sugar and TG or complicated with vascular disease, the LPO reaction enhanced markedly. BSHX tablets have the function of reducing the serum LPO and blood sugar, clearing cholesterol. Therefore, this tablet will have a good prospect in the prevention and treatment for DM patients complicated with vascular disease. PMID- 8732152 TI - [Effect of tetramethylpyrazine on expression of collagen genes in primary cultured smooth muscle cells in rats]. AB - For the purpose of studying the mechanism of tetramethylpyrazine (TMP) to prevent the restenosis after percutaneous transluminal coronary angioplasty (PTCA) and promote blood circulation to remove stasis. 40 micrograms of TMP was used to treat cultured vascular smooth muscle cells (VSMs). The total RNA was extracted from treated VSMC. The RNA Northern blot hybridization with procollagen genes alpha i-I and alpha 1-III probes was used to investigate its change in mRNA level. The results indicated that TMP could inhibit significantly the transcription of procollagen genes alpha 1-I and alpha 1-III. PMID- 8732153 TI - [Effects of qiwei baizhu san in inhibiting replication of human rotavirus in vitro]. AB - Qiwei Baizhu San was found to have an inhibitory effect on human rotavirus (HRV)at monolayer of MA104 Cells. A 50% reduction in plaque number, a 10(1.86)TCID50 decrease in viral replication index and around 60% inhibition in viral RNA synthesis were observed at concentration of 100 mg/ml. Furthermore, the cytotoxicity of the decoction was low, while its promoting effect on growth and proliferation of the culture cells was observed at the concentrations of 12.5-50 mg/ml. The decoction was also found to have effects in prolonging the survival time of HRV infected cells and promoting the regeneration of the infected cells. PMID- 8732154 TI - [Effect of er-xian decoction and its disassembled prescription on enzyme activities and their gene expression of antioxidant enzymes in aging rat]. AB - In order to study the mechanism of aging, this experiment adopted molecular biological techniques to observe the effect of Er-Xian decoction (EXD)and its disassembled prescription on the enzyme activities and their gene expression of superoxide dismutase (SOD)and catalase (CAT). The results showed that in old rat, the amount of lipid peroxide (LPO) increased significantly; the enzyme activities such as SOD, CAT decreased significantly; and the lowering of these activities had a direct bearing on the gene expression level of their enzyme protein. After treatment by EXD and its disassembled prescription in aging rat, the amounts of LPO reduced significantly, the activities of SOD, CAT enhanced significantly. Moreover, it was observed that the increasing of these enzyme activities was related to the enhancing of their gene expression. It was suggested that EXD had the effect of anti-aging in that it could raise the activities of antioxidant enzymes and reduce the product of free radicals by means of regulating and enhancing the gene expression levels of their enzyme protein. PMID- 8732155 TI - [Developing the advantage of Chinese integrated medicine--review of "Clinical effective cases of traditional Chinese medcine"]. PMID- 8732156 TI - [Pharmacology of Sophora flavescense and its clinical uses]. PMID- 8732157 TI - [Research on plasma TXA2, PGI2 levels, blood stasis syndrome and promoting blood circulation]. PMID- 8732158 TI - Gene marking. PMID- 8732159 TI - Targeting of retroviral vectors through protease-substrate interactions. AB - Targetable, injectable vectors would greatly facilitate the development of in vivo therapy strategies. Viral and nonviral vectors can be targeted through ligand-receptor interactions, but protease-substrate interactions have not previously been exploited for vector targeting. Epidermal growth factor (EGF) was fused to a retroviral envelope glycoprotein via a cleavable linker comprising a factor Xa protease recognition signal. Vector particles displaying the cleavable EGF domain could bind to EGF receptors on human cells but did not transfer their genes until they were cleaved by factor Xa protease, whereupon gene delivery proceeded normally. Proteolytic activation of receptor-targeted vectors can therefore provide the basis for a novel two-step targeting strategy that may facilitate efficient targeted in vivo delivery of therapeutic genes. PMID- 8732160 TI - Efficient delivery of triplex forming oligonucleotides to tumor cells by adenovirus-polylysine complexes. AB - Oligonucleotides (ODNs) show great promise in their ability to specifically inhibit single gene expression but must cross the cell membrane, escape the endosomal vesicle, and possibly traverse the nuclear membrane to arrive at their intracellular target molecules. In an attempt to improve the delivery of phosphodiester triplex forming ODNs to malignant cells, we have constructed adenovirus-polylysine (AdpL)-ODN complexes designed to take advantage of the receptor mediated endocytosis of adenoviruses to transfer the ODNs to the cell nucleus. Treatment of several different types of tumor cells in culture by AdpL ODN complex resulted in superior uptake and persistence of the ODNs compared to both free ODN and cationic lipid-ODN complexes. Nuclear uptake peaks at 4 h and intact ODN persists in the nucleus with a half-life of 12 h. ODN concentrations of 20-70 microM are achieved at 24 h in all monolayer cell lines evaluated to date. ODNs are detected in 50-100% of the total cell population by immunohistochemistry with apparent uptake into vesicles and nuclear localization. Luciferase expression of a co-delivered reporter plasmid suggests that these ODNs are free in the nucleus. AdpL-ODN complexes will provide a valuable tool for delivering unmodified ODNs to the nucleus of malignant cells. PMID- 8732161 TI - In vivo adenovirus-mediated gene transfer into normal and cystic rat kidneys. AB - Gene transfer into the mammalian kidney has proved difficult because of the structural complexity of the organ and its low mitotic index. This article describes the use of intra-arterially injected adenovirus to study gene transfer into the rat kidney in vivo. By pre-chilling the kidney, and incubating the virus with the kidney in the cold for extended periods of time, we were able to successfully transfer a beta-galactosidase (beta-gal) reporter gene into the vasculature without ischemic injury to the kidney. Transfer occurred largely in the cortex when cold was used alone, whereas with the use of cold and vasodilators, transfer was accomplished into the outer medulla in both the inner and outer stripes. In the Han:SPRD rat model of autosomal dominant polycystic kidney disease (ADPKD), gene transfer occurred into the vasculature, some epithelial cysts and interstitial cells. This is the first description of substantial in vivo gene transfer into both normal and cystic kidneys. The methodology could find application in the creation of new models of renal disease, for in vivo therapeutic intervention or for genetic modification of an allograft at the time of harvest. PMID- 8732162 TI - Gene therapy in the adult primate brain: intraparenchymal grafts of cells genetically modified to produce nerve growth factor prevent cholinergic neuronal degeneration. AB - Gene therapy may be a useful means of delivering substances to the brain that are capable of preventing neuronal degeneration. In the present experiment, we determined whether intraparenchymal transplants of primary autologous cells genetically modified to produce nerve growth factor (NGF) would prevent injury induced degeneration of cholinergic neurons. Cultured primary monkey fibroblasts were genetically modified to produce human NGF, and secreted 13.2 ng NGF/10(6) cells/h in vitro. Adult monkeys then underwent fornix transections to induce degeneration of basal forebrain cholinergic neurons, and received autologous grafts of either NGF-producing or control, beta-galactosidase-producing fibroblasts directly into the basal forebrain region. One month later, 61.7 +/- 8.9% of cholinergic neurons remained indentifiable in NGF-graft recipients compared to 26.2 +/- 5.0% in control graft recipients (P < 0.02). Neuronal protection correlated with the accuracy of graft placement: up to 92% protection from neuronal degeneration occurred when NGF-secreting grafts were accurately placed immediately adjacent to injured neurons. Thus, intraparenchymal NGF delivery to the adult primate brain by gene transfer can prevent the degeneration of basal forebrain cholinergic neurons. Gene therapy can target intraparenchymal brain sites for regionally specific neurotrophin delivery, thereby avoiding limitations imposed by diffusion of substances across the blood-brain barrier and through CNS parenchyma, while avoiding adverse effects of neurotrophic factors delivered in a non-directed manner to the central nervous system. The delivery of NGF by gene transfer to the brain merits further study as a means of preventing cholinergic neuronal degeneration in human disorders such as Alzheimer's disease. PMID- 8732163 TI - Gene therapy of rat C6 glioma using adenovirus-mediated transfer of the herpes simplex virus thymidine kinase gene: long-term follow-up by magnetic resonance imaging. AB - The herpes simplex virus thymidine kinase gene was transferred into C6 glioma cells by infection with a recombinant adenovirus. In vitro, a 10 microM ganciclovir concentration was able to kill 100% of the infected cells. For in vivo experiments, brain tumors were established by stereotactic injection of C6 glioma cells in the caudate nucleus of rats. Five days later, the recombinant adenovirus was inoculated into the tumors and the animals were treated by intraperitoneal injections of ganciclovir for 14 days. At the end of ganciclovir therapy, histological examination revealed a 28-fold decrease in tumor volumes in the treated animals, as compared with control animals. In long-term studies, the mean survival time of the treated animals were four-fold longer than that of control ones. Magnetic resonance imaging demonstrated an apparent complete tumor regression in 62% of the animals. However, late tumor recurrence was observed in the treated animals. Repeated inoculation of C6 glioma cells in the contralateral hemisphere of long-term surviving animals resulted in either tumor rejection or slowly growing tumors. These findings demonstrate the potential efficacy of adenovirus-mediated transfer of the herpes simplex virus thymidine kinase gene and ganciclovir administration in the treatment of rat gliomas. PMID- 8732164 TI - Cell cycle state, response to hemopoietic growth factors and retroviral vector mediated transduction of human hemopoietic stem cells. AB - A multi-parameter fluorometric analysis was applied to study in vitro proliferation and expansion of a candidate hemopoietic stem cell population, ie CD34brightLin cells. In primary hemopoietic cell samples the CD34brightLin population comprised on average, 1% of CD34+ cells from bone marrow (BM) or umbilical cord blood and 0.1% of mobilized peripheral blood CD34+ cells. The fraction of CD34brightLin cells engaged in the S+G2/M phases of the cell cycle was largest in regenerating BM (10.6% versus 1-2% in the other sources). Maintenance of CD34+ BM cells in hemopoietic growth factor supplemented liquid cultures resulted in a decrease of the CD34brightLin population in most samples. Cell cycle analysis showed the constant existence of proliferating CD34+ cells during the culture period. The fraction of cycling CD34brightLin cells was, however, very small and only occasionally exceeded input values. At all tested time-points during culture, BM CD34+ cells could be transduced by a single incubation with a recombinant retrovirus supernatant. CD34brightLin cells, however, were much more refractory to retroviral vector-mediated transduction. This could be explained only in part by quiescence of CD34brightLin cells. Hence, factors other than cell proliferation clearly influence the early stages of retroviral transduction of human hemopoietic stem cells. PMID- 8732165 TI - Expression of chimeric envelope proteins in helper cell lines and integration into Moloney murine leukemia virus particles. AB - New retroviral constructs with a grafted specificity of infection could become useful gene delivery vehicles with applications in systemic gene therapy. We have constructed retroviral vectors to target gene transfer to human tumor cells. Chimeric envelope proteins have been expressed to obtain viral particles with a defined specificity of infection. Two tumor cell-specific recognition domains were cloned and fused with the viral envelope gene. A recognition domain specific for ErbB-2 expressing tumor cells was derived from a monoclonal antibody directed against the ErbB-2 receptor in the form of a single chain antibody domain (scFv erbB-2). The receptor binding domain was derived from the heregulin gene (HRG70). This domain provides recognition specificity for ErbB-3 and ErbB-4 receptor expressing tumor cells. The recognition domains were inserted at the amino terminal end into the MoMLV envelope gene. Helper cell lines were established which express the recombinant envelope protein genes, the gag and pol genes and packageable retroviral RNA. The analysis of the helper cell line revealed that the recombinant ErbB-2 scFv-envelope protein was expressed, but not incorporated into viral particles. The scFv-erbB-2 envelope protein was not inserted into the cell membrane and the assembly of retroviral particles was not completed. In contrast, the HRG70-envelope protein was expressed on the surface of the helper cells and incorporated into retroviral particles. The HRG70-envelope protein, however, did not alter the host range of infection. Only cells expressing the ecotropic viral receptor could be infected. PMID- 8732166 TI - Retroviral gene transfer and sustained expression of human arylsulfatase A. AB - Transduction of mouse hematopoietic stem cells and their progeny was studied using a recombinant retroviral vector (MFG-ASA) which incorporates the human arylsulfatase A gene (ASA; EC 3.1.6.8). Successful transduction was demonstrated in spleen colonies of mice that received bone marrow transplantation, cultured bone marrow-derived macrophages, visceral tissues and brain of long-term reconstituted mice, and also the spleen colonies of secondarily transplanted mice. The efficiency of transduction in primary spleen colonies was 90%. Expression of the ASA transgene exceeded endogenous levels in spleen colonies and in cultured macrophages by 50-100%. Enzyme activity in the visceral tissues of long-term reconstituted mice consistently showed elevated ASA activity, greater than three-fold in the spleen and lung of one animal. Increased activity of ASA also could be detected in secondary spleen colonies. These data demonstrate the usefulness of the MFG-ASA vector for efficient gene transfer and expression in mouse hematopoietic stem cells and their differentiated progeny. The presence of vector DNA in the brain 4 months after transplantation suggests a role for gene transfer and stem cell transplantation in the treatment strategies for metachromatic leukodystrophy. PMID- 8732168 TI - Expression of mutant and wild-type gag proteins for gene therapy in HIV-1 infection. AB - The effect of expression of defective HIV-based retroviral constructs in CD4 positive lymphocytes on subsequent infection of the cell by HIV-1 was studied. A vector containing a N terminally elongated gag protein which was noncleavable and myristoylation negative was not effective at inhibiting HIV assembly or viral replication in the culture. Expression of a wild-type HIV gag in trans led to an increase in cytopathicity within the culture such that all the cells died. A control LTR containing vector had no effect. A myristoylation negative gag would not appear to be a useful dominant negative inhibitor of HIV replication, but might be usable as a post-exposure immunogen. Post-infective immunisation with wild-type HIV-1 gag would appear to risk increasing virus-related cytopathicity. PMID- 8732167 TI - Novel, high expressing and antibiotic-controlled plasmid vectors designed for use in gene therapy. AB - The promise of effective gene therapy can only be accomplished by high-level expression and regulatable delivery of gene products. To achieve this end, a eukaryotic expression plasmid was modified to make transcription dependent on a tetracycline(Tc)-regulated chimeric transactivator. Mouse muscle injected with this two plasmid cis/trans control system expressed reporter proteins at levels five- to 10-fold greater than the cytomegalovirus immediate-early promoter controlled parental plasmid. Tetracycline could be useful to either repress or activate transactivator-controlled expression based on the position of the tetO control sequences within the reporter plasmid. Finally, a prototype single plasmid construct was made and shown to express a self-regulating bicistronic transcript containing both the reporter and the transactivator. These Tc controlled plasmids, termed maximum expression and regulated vectors (MERVs), have the potential to target a variety of gene therapy applications. PMID- 8732169 TI - Gene transfer into haemopoietic cells: a challenge for gene therapy--European Concerted Action, Workshop--Marseille, 18-19 October 1995. PMID- 8732170 TI - Expression levels by retroviral vectors based upon the N2 and the MFG backbones. PMID- 8732171 TI - Clinical relevance of hepatitis C virus quasispecies. AB - It has been shown that hepatitis C virus (HCV) populations in infected individuals are composed of quasispecies with diverse mutations. The analysis of these variants may reveal mechanisms of the persistence of HCV infection, carcinogenesis and resistance to antiviral therapy. Recently, genetic features of interferon-resistant HCV have been elucidated through the analysis of interferon resistant quasispecies, making it possible to predict interferon efficacy by detecting interferon-resistant strains. PMID- 8732172 TI - Targeted delivery of antisense DNA in woodchuck hepatitis virus-infected woodchucks. AB - An asialoglycoprotein-based DNA delivery system containing an antisense oligo DNA against the polyadenylation region and adjacent upstream sequences of woodchuck hepatitis virus (WHV) was prepared. Experimental woodchucks were inoculated neonatally with the woodchuck virus 23 weeks before initiating the study, and all animals subsequently developed hepatitis as evidenced by the presence of measurable levels of circulating viral DNA. Animals were injected intravenously (i.v.) with asialoorosomucoid (AsOR)-poly-L-lysine complexes containing 0.1 mg kg 1 antisense DNA for five consecutive days. Levels of surface antigen did not differ substantially between treated and control animals. However, intravenous administration of complexed antisense DNA significantly decreased viraemia, as shown by a five- to 10-fold decrease in circulating viral DNA 25 days post treatment. The decline lasted for at least 2 weeks, after which there was a gradual increase in DNA levels. Antisense DNA alone or a complex containing a random oligo DNA of the same size and linkage failed to have any significant effect on viral DNA levels. We conclude that antisense oligo DNA can be targeted to the liver in vivo, resulting in a substantial and prolonged decrease in viral DNA levels in WHV-infected woodchucks. PMID- 8732173 TI - Analysis of hepatitis B virus precore variants in hepatitis B e antibody-positive patients treated with prednisone plus interferon. AB - To assess the effects of prednisone and interferon on the distribution of hepatitis B virus (HBV) precore mutants, nine hepatitis B e antibody (HBeAb) positive patients with HBV chronic infection were studied. Patients were treated with prednisone (30 mg day-1 for 4 weeks, followed by 20 mg day-1 for 2 weeks and by 10 mg day-1 for 1 week), followed by recombinant interferon-alpha (15 MU thrice per week) for 6 months, without a clearance period. The HBV precore region was amplified by polymerase chain reaction (PCR) and distribution of the precore mutants was determined by hybridization of PCR products. Moreover, the glucocorticoid-responsive element (GRE) was sequenced to determine whether changes in the sequence were produced at the end of prednisone treatment. During prednisone treatment, changes in alanine transaminase (ALT) were observed in only two patients, in who ALT decreased to nearly normal values. In three patients ALT normalized at the end of interferon treatment. At baseline, wild-type HBV alone was detected in one patient, while seven patients were infected by a mixture of wild-type and precore mutants, predominantly wild type. At the end of prednisone treatment, two patients were infected by only wild-type HBV. The proportion of precore mutants decreased in three cases, while no changes were observed in three. At the end of interferon treatment, the precore mutant proportion decreased in the three responders, while tending to increase or remain unchanged in the rest. No significant changes in GRE sequence were found as a result of prednisone treatment. Our results would appear to confirm the role of the immune system in the selection of precore mutants. PMID- 8732174 TI - Variations of hepatitis C virus NS5B sequence (nucleotides 8261-8566) do not correlate with response to interferon-alpha therapy. AB - Hepatitis C virus (HCV) is an RNA virus with the NS5B gene encoding an RNA dependent RNA polymerase. Interferon-alpha (IFN-alpha) is effective against HCV and its effect is believed to be related to its antiviral activity. To determine whether sequence variations of the HCV NS5B region correlate with response to IFN therapy, pretreatment serum samples from 40 patients with chronic HCV infection who were subsequently treated with IFN (> or = 3 MU thrice weekly for 24 weeks) and had well-characterized biochemical responses were studied. Reverse transcription-polymerase chain reaction (RT-PCR) was performed to generate an approximately 365-bp fragment from which nucleotide sequence and genotypes were determined. By comparing the nucleotide sequences and the encoded amino acid sequences of samples from each group, no response group-specific nucleotide or encoded amino acid substitution was identified. Most of the substitutions identified were synonymous (usually by changes at the third position of the codon). These data suggest that these substitutions were selective rather than spontaneous events. Of the few non-synonymous substitutions identified, none was correlated with subsequent response to IFN, either within or across genotypes. PMID- 8732175 TI - Hepatitis C virus genotypes in patients with chronic liver disease and haemodialysis patients from Saudi Arabia. AB - The genotypes of hepatitis C virus (HCV) were investigated in 28 Saudi patients (21 males, seven females; age range 23-68 years; mean 45.0 years) with histologically proven chronic hepatitis (13 chronic active hepatitis and 15 liver cirrhosis) and in 32 Saudi patients with chronic renal failure maintained on haemodialysis (22 males, 10 females; age range 18-60 years; mean 40.0 years) who also had liver disease due to HCV. Among the 28 patients with chronic liver disease genotype 4 was the predominant one (60.7%), followed by types 1b (21.4%), 1a (14.3%) and 2a (3.6%). The distribution of genotypes was similar in patients with chronic active hepatitis to those with liver cirrhosis. Among the 32 patients with chronic renal failure and maintained on haemodialysis, genotype 4 was also the dominant type (55.0%), followed by 1a (25.0%), 1b (21.9%) and 2a (3.1%). In all categories studied the prevalence of genotypes between males and females was the same. As our patients were selected from various regions of Saudi Arabia, we believe that genotype 4 is the predominant one throughout the whole kingdom. PMID- 8732176 TI - The distribution of hepatitis C virus genotypes in Turkish patients. AB - The distribution of hepatitis C virus (HCV) genotypes was investigated in 89 HCV infected Turkish patients. Blood samples were collected from haemodialysis patients (n = 45), chronic liver disease (CLD) patients (n = 38), acute non-A, non-B (NANB) hepatitis patients (n = 2) and blood donors (n = 4). HCV RNA sequences were amplified in the 5' non-coding region and were typed by restriction fragment length polymorphism analysis. The predominant genotype was 1b (75.3%), followed by 1a (19.1%), 2 (3.4%) and 4 (2.2%). While there was no significant difference in the distribution of HCV genotypes with respect to age, sex, transfusion history, alanine aminotransferase levels or liver histology (in the CLD group), type 1a-infected patients were younger than type 1b-infected patients (P < 0.05) in the haemodialysis group. Serological reactivity to recombinant HCV proteins was assessed in 58 samples using the Chiron RIBA-2 assay. The reactivity of samples from patients infected with type 1b with 5-1-1 and c100 antigens was significantly lower (P < 0.05) than the reactivity of samples from those infected with type 1a. These results, together with the results of two previous studies, indicate that HCV genotypes 1, 2, 3 and 4 are prevalent in different frequencies in the Turkish population. Determination of the genotype distribution of HCV in a geographical area may provide important clues for studying the epidemiology, transmission and pathogenesis of HCV-related diseases and may also aid in improving serological assays to detect HCV infection. PMID- 8732178 TI - What's in a name? PMID- 8732177 TI - Clinical and serological courses of a newborn with post-transfusion hepatitis C. AB - The clinical and serological course of a haemophilic baby who was transfused with 160 ml of blood containing the hepatitis C virus (HCV) (0.70 Meq. ml-1) on the sixth-post-natal day is described. He is the infant of an HCV-negative mother. One month after the transfusion, there was a marked increase in HCV RNA and a small amount of HCV antibody was detected. This case provides evidence that a newborn is capable of producing HCV antibodies. PMID- 8732179 TI - Quantification of biomedical NMR data using artificial neural network analysis: lipoprotein lipid profiles from 1H NMR data of human plasma. AB - Artificial neural network (ANN) analysis is a new technique in NMR spectroscopy. It is very often considered only as an efficient "black-box' tool for data classification, but we emphasize here that ANN analysis is also powerful for data quantification. The possibility of finding out the biochemical rationale controlling the ANN outputs is presented and discussed. Furthermore, the characteristics of ANN analysis, as applied to plasma lipoprotein lipid quantification, are compared to those of sophisticated lineshape fitting (LF) analysis. The performance of LF in this particular application is shown to be less satisfactory when compared to neural networks. The lipoprotein lipid quantification represents a regular clinical need and serves as a good example of an NMR spectroscopic case of extreme signal overlap. The ANN analysis enables quantification of lipids in very low, intermediate, low and high density lipoprotein (VLDL, IDL, LDL and HDL, respectively) fractions directly from a 1H NMR spectrum of a plasma sample in < 1 h. The ANN extension presented is believed to increase the value of the 1H NMR based lipoprotein quantification to the point that it could be the method of choice in some advanced research settings. Furthermore, the excellent quantification performance of the ANN analysis, demonstrated in this study, serves as an indication of the broad potential of neural networks in biomedical NMR. PMID- 8732180 TI - T1 and T2 relaxation times of the major 1H-containing metabolites in rat brain after focal ischemia. AB - The relaxation properties of water and metabolites were measured in rat brain following the occlusion of the middle cerebral artery (MCA) with localized 1H MRS. The PRESS sequence was employed to select volumes of 39 microL in the ischemic and the contralateral hemisphere. T1 and T2 relaxation times and peak intensities of water, choline containing compounds (Cho), creatine and phosphocreatine (Cre) and N-acetyl aspartate (NAA) in both hemispheres were determined at 3-6 h, 1 day and 3 or 4 days after occlusion. Lactate in the ischemic hemisphere was also quantified. The relaxation properties and peak intensities of NAA, Cre and Cho remained unchanged in the ischemic volume during the first 3-6 h of ischemia as compared to the contralateral volume. Water T2 was slightly increased in the ischemic volume. After 24 h the T1 and T2 of water and Cre and the T1 of Cho had increased significantly in the ischemic volume, while the peak intensities of Cho, Cre and NAA were reduced. It appears therefore that tissue changes which occur in the early phase of ischemia have no significant effects on the relaxation behaviour of the metabolites. However, ischemic brain damage affects the relaxation behaviour and concentration of the metabolites and water at later stages. PMID- 8732181 TI - Characteristic metabolic profiles revealed by 1H NMR spectroscopy for three types of human brain and nervous system tumours. AB - Cell culture techniques, high-resolution in vitro 1H NMR spectroscopy, and chromatographic analyses were used to compare the properties of three types of human brain and nervous system tumours. Cell lines were immunocytochemically characterized at all stages in culture with specific antibodies. Intracellular metabolites present in cell extracts were analysed by 1H NMR spectroscopy and by high performance liquid chromatography (HPLC). The spectra from meningiomas, neuroblastomas, and glioblastomas displayed, in addition to similarities including the presence of signals from leucine, isoleucine, valine, threonine, lactate, acetate, glutamate, choline-containing compounds and glycine-certain distinguishing metabolic features. Spectra from meningiomas featured relatively high signals from alanine. Intense signals from creatine were present in neuroblastoma spectra, while in spectra from glioblastoma they were not detectable. We found statistically significant differences by 1H NMR spectroscopy in the amounts of alanine, glutamate, creatine, phosphorylcholine and threonine among the types of tumours examined. HPLC determinations confirmed that there were also other metabolites specific to a type of tumour, such as taurine, gamma aminobutyric acid, and serine. We suggest that these findings have potential relevance for the development of non-invasive diagnosis of tumour lineage by 1H NMR spectroscopy in vivo. PMID- 8732182 TI - Dynamic monitoring of cerebral metabolites during and after transient global ischemia in rats by quantitative proton NMR spectroscopy in vivo. AB - Localized proton NMR spectroscopy was used to dynamically monitor alterations of cerebral metabolites before, during, and after a 10 min period of global forebrain ischemia in anesthetized rats. Metabolic assessment was based on user independent determination of absolute brain concentrations at a nominal temporal resolution of 1.6 min. While the concentrations of N-acetyl aspartate (neuronal marker), creatines, cholines, and myo-inositol (glial marker) remained constant, ischemia induced a rapid decline of brain glucose. One hour after reperfusion, glucose recovered to 4.1 +/- 2.2 mmol/kg wet weight significantly above the basal value of 2.3 +/- 1.3 mmol/kg wet weight. Mirroring glucose depletion, lactate increased from 1.0 +/- 0.6 to 13.5 +/- 1.5 mmol/kg wet weight 10-15 min after the onset of ischemia. During reperfusion lactate clearance was characterized by a first-order rate constant of 0.03/min. The time courses of glucose and lactate reflect the rapid onset of anaerobic glycolysis during states of critically diminished blood flow. Assuming complete ischemia the production of lactate from glucose and cerebral glycogen stores yields a brain glycogen concentration of 4.7 +/- 0.9 mmol glycosyl unit/kg wet weight. Elevation of brain glucose during early reperfusion suggests a transient mismatch of glucose uptake and consumption during the first 1-2 hours post ischemia. PMID- 8732183 TI - Signal profile measurements for evaluation of the volume-selection performance of ISIS. AB - High-resolution signal profiles obtained with a test phantom were used in this study to evaluate the volume-selection performance of an implementation of ISIS (Image Selected In vivo Spectroscopy). The phantom simulated the brain with regard to volume and loading of coil. A remotely controlled, movable signal source inside the phantom was filled with orthophosphoric acid. Signal profiles of the volume of interest (VOI) were measured in three perpendicular directions. Special interest was focused on the transition zones, the position of the profiles, and the effects of off-resonance and T1 smearing. The transition zones were on average 5.6 mm wide and the full width at half maximum (FWHM) was 35 mm for a VOI of 40 x 40 x 40 mm3. The positions of the centre of the signal profiles were x = 3.2, y = -0.7 and z = 3.3 mm off-centre. The deviation of the volume position could be explained by off-resonance effects during imaging and spectroscopy. These data illustrate the importance of detailed knowledge of the volume-selection performance when attempting precision measurements using image guided in vivo MRS. PMID- 8732184 TI - Glial cell transplantation and remyelination of the central nervous system. AB - Glial cell transplantation has proved to be a powerful tool in the study of glial cell biology. The extent of myelination achieved by transplanting myelin producing cells into the CNS of myelin mutants, or into focal demyelinating lesions has raised hope that such a strategy may have therapeutic applications. Oligodendrocytes or Schwann cells could be used for repair. It is likely that the immature stages of the oligodendrocyte lineage have the best phenotypic characteristics for remyelination when transplanted, either as primary cells or as immortalized cells or cell lines. Prior culturing and growth factor treatment provides opportunities to expand cell populations before transplantation as dissociated cell preparations. Cell lines are attractive candidates for transplantation, but the risk of transformation must be monitored. The application of this technique to human myelin disorders may require proof that migration, division and stable remyelination of axons by the transplanted cells can occur in the presence of gliosis and inflammation. PMID- 8732186 TI - Telencephalic transplants in mice: characterization of growth and differentiation patterns. AB - Telencephalic grafting represents a powerful tool for developmental studies and for the investigation of biological features of transgenic brain tissue. The interpretation of grafting experiments, however, requires detailed knowledge of graft biology. Therefore, we have characterized growth rates, graft size, and differentiation of embryonic telencephalic tissue harvested at various developmental stages and grafted into the caudoputamen and lateral ventricles of histocompatible mice. A total of 164 grafts were analysed up to 500 days after transplantation. Of all transplants, 79.3% resulted in the formation of solid neural grafts. Grafted cells were identified by 3H-thymidine labelling and autoradiography. Proliferation was studied by bromodeoxyuridine incorporation and decreased from an initial 35% at 1-3 d after grafting to less than 1.6% after 40 days. The graft size was measured as a function of the embryonic age of the transplanted tissue. Our data indicate that telencephalic tissue harvested at embryonic day E 12.5 reproducibly yields large, fully differentiated neuroectodermal grafts. The parameters defined in this study will be useful for detailed analysis of neuroectodermal tissue from mice undergoing fatal neurodegeneration, such as knockout mice bearing lethal mutations. PMID- 8732185 TI - The expression of the endothelial cell antigen CD34 in demyelinating disease. AB - In this study, the antigenic expression of CD34, a 110 kDa glycoprotein which is expressed on human haemopoietic progenitor cells and vascular endothelium, has been assessed in a variety of neuropathological conditions, including infectious and demyelinating disease. Using immunoperoxidase staining on paraffin sections, the immunohistochemical results show that CD34 antigen is expressed widely on human CNS endothelium in grey and white matter, in the eye including retina, and in the anterior and posterior lobes of the pituitary. In demyelinating disease CD34 antigen expression was not detected in acute lesions, whereas strong expression was observed in old lesions. CD34 endothelial positivity was observed in areas of gliosis, vasogenic oedema, vascular disease and in Alzheimer's and Parkinson's disease pathology. A general pattern emerged, with CD34 antigen reactivity predominantly negative in areas of inflammation with demyelination but positive in adjacent non-inflamed tissue, irrespective of myelin pathology. We conclude that perivascular inflammation is a key factor in the absence of immunoreactivity of CD34 in the CNS in demyelinating disease. PMID- 8732187 TI - Comparative in vivo and pathological analysis of the blood-brain barrier in mouse telencephalic transplants. AB - The post-transplantation status of the blood-brain barrier (BBB) is still a matter of debate. In an attempt to define BBB properties after neural transplantation in mice of a defined genetic background, we have used two exogenous markers (horseradish peroxidase and Evans blue), one endogenous marker (immunoglobulins), and in vivo contrast enhanced magnetic resonance imaging (MRI) and compared the results obtained with the different methods. With all four techniques employed, we found the BBB to be reconstituted in 67% of the grafts 3 weeks after grafting, and in more than 90% of all grafts 50 days after grafting. Horseradish peroxidase and contrast enhanced MRI were the most sensitive techniques, the latter offering the unique advantage of repetitive scanning of individual grafts. Our findings provide important information for transplantation studies in mouse models for neurodegenerative diseases. PMID- 8732188 TI - Aberrant Timm-stained fibres in the dentate gyrus following tetanus toxin-induced seizures in the rat. AB - The present study addresses whether seizures, which result from the chronic block of inhibition caused by an intrahippocampal injection of tetanus toxin, induce axonal sprouting of the hippocampal mossy fibres. Timm stain was used to identify the mossy fibre terminals. In nine of 15 animals killed at 1 month or later after an injection of tetanus toxin, Timm-stained terminals were observed bilaterally in the inner molecular layer and in seven animals a meshwork of Timm-stained fibres/ terminals was also observed bilaterally in the outer molecular layer of the fascia dentata. None of these changes were observed in any of the 12 saline injected controls. There was no obvious correlation between the number of motor fits an animal exhibited and the amount of Timm-stained fibre sprouting present in either the inner or outer molecular layer. The Timm-stained axonal sprouting into the outer molecular layer of the fascia dentata may simply reflect the reinnervation of sites on the granule cell dendrites, previously occupied by the terminals of the hilar somatostatin-containing cells. These hilar somatostatin containing cells which are believed to project to the outer molecular layer are known to succumb to the seizure activity in this animal model of epilepsy. PMID- 8732190 TI - Mass slaughter policy 'wholly unreal' says agriculture minister. PMID- 8732189 TI - Astrocytes in the hypoglossal nuclei of sudden infant death syndrome (SIDS) infants: a quantitative study. AB - It has been suggested that brain stem hypoxia or ischaemia underlies the sudden infant death syndrome (SIDS), but previous reports of astrocytosis in the brain stems of SIDS infants have been contradictory. A volumetric quantitative technique was, therefore, developed to compare astrocyte numbers and sizes in the hypoglossal nuclei of SIDS and control infants. In 12 SIDS and eight control infants, serial sagittal sections were taken through the hypoglossal nucleus and every tenth section was stained for glial fibrillary acidic protein. Astrocytes were counted in the central 4% of a grid stepped throughout the hypoglossal nucleus, and the heights of 100 astrocyte nuclei were measured with a microcator. Astrocyte number, corrected for section thickness and nuclear height, was divided by the volume of the hypoglossal nucleus to calculate astrocyte density. Numbers of astrocytes did not differ significantly between SIDS (mean number 44729, SD 12096) and control (mean number 46562, SD 11060) infants. Astrocyte nuclear height did not differ significantly between groups (SIDS: mean height 3.98 microns, SD 0.22; control: mean height 3.84 microns, SD 0.31). Astrocyte density was similar in SIDS (mean density 24378 astrocytes/mm3, SD 6155) and control (mean density 23978 astrocytes/mm3, SD 4031) infants. No quantitative evidence of astrocytosis was found in the hypoglossal nuclei of SIDS infants. This implies that SIDS infants die without previous episodes of hypoxia/ischaemia severe enough to damage the brain stem. PMID- 8732191 TI - Castration of calves: a study of methods used by farmers in the United Kingdom. AB - A postal survey of farmers was conducted to determine the main methods used to castrate calves, and by whom and how they were applied. Among the 28 per cent of farmers who replied, those who did castrate calves used one or more of three methods: the Burdizzo was used by 43 per cent of farmers, surgery by 39 per cent, and rubber rings by 32 per cent, with 10 per cent using more than one method. Calves were castrated at all ages from less than one week to over six months, with one third of them being castrated at an age that legally requires the operation to be done under local anaesthesia by a veterinary surgeon. Rubber rings were never used by veterinary surgeons, but they carried out 43 per cent of surgical castrations, which was the method of choice in older calves. Local anaesthetic was used on 15 per cent of farms, mainly for surgical castrations. Sixty-seven per cent of farmers using the Burdizzo applied it twice, with the majority correctly applying the second crush below the first, and 90 per cent used precautions to control infection after surgical castration. PMID- 8732192 TI - Epidemiology of subclinical ovine psoroptic otoacariasis in Great Britain. AB - Seven sheep flocks with subclinical psoroptic otoacariasis were investigated. Psoroptes species mites were isolated from 3.1 per cent of the 2676 sheep examined and the prevalence in the flocks ranged between 1.3 and 23.9 per cent, with the highest infestations in pedigree flocks. Unilateral or bilateral infestations were found in sheep of all ages. Adult and shearling rams, with prevalences of 21.5 and 14.2 per cent, respectively, were the most affected and two lambs, two and eight days old, were the youngest to be infested. There was no evidence of vertical transmission. The infestation affected several breeds of sheep. No other known hosts for Psoroptes species present on the premises were shown to be infested. Non-parasitic for-age mites were also isolated from the ear canals of the sheep and may have contributed to the clinical signs which, in adult sheep, ranged from aural haematomas/fibrosis (cauliflower ears), and violent head shaking and ear rubbing leading to excoriation and wounding of the ear and base of the ear. The signs in lambs included plaques of scab (often bloody) on the external ear cleft, excoriation of the base of the ear, ear scratching with the hind feet and inflammation of the external aspects of the horizontal canal. In all cases the internal pinnae were clear of typical psoroptic scabs. Psoroptes mites were isolated from 28.6 per cent of the damaged ears and from 7.8 per cent of the undamaged ears. There was no evidence of classical sheep scab in any of the flocks. Plunge dipping in diazinon, propetamphos or flumethrin, or the use of synthetic pyrethroid pour-on preparations and an oral drench of ivermectin had little effect because ear mite infestations were detected in the flocks after these treatments. An injection of ivermectin at 200 micrograms/kg bodyweight effectively eradicated the infestations when it was administered to these infested flocks. PMID- 8732193 TI - Evaluation of a one-step test for rapid, in practice detection of rotavirus in farm animals. AB - An immunochromatographic test for the detection of group A rotavirus was evaluated against a reference group A rotavirus ELISA, by using a panel of 161 bovine, porcine and equine faecal samples submitted for routine examination. The sensitivity of the test was 89 per cent and the specificity 99 per cent compared with the ELISA. Its reproducibility was 100 per cent. The simplicity and rapidity of the test procedure make it suitable for use in practice. PMID- 8732194 TI - Paradoxic glucosuria (Fanconi syndrome) in a bull. PMID- 8732195 TI - Developmental abnormalities of the reproductive tract associated with infertility in Highland heifers. PMID- 8732196 TI - Parapoxvirus infection in red squirrels. PMID- 8732197 TI - Vero cytotoxin-producing Escherichia coli O157 associated with companion animals. PMID- 8732198 TI - Autoimmunity in the peripheral nervous system. AB - The PNS is an immunocompetent organ. The participating cellular and humoral elements in the local immune circuitry have been identified. The model disorder EAN is extensively being used to examine the induction, amplification and effector phase of autoimmune responses to peripheral nerve antigens (Figure 9). Potential autoantigens contained in the myelin sheath and on the axolemma have been characterized. Recent years have seen a rapid growth of information concerning the pathogenesis of the Guillain-Barre syndrome, which has turned out to be a heterogeneous disorder, both in clinical characteristics, course and prognosis, and in the underlying pathology. It can be anticipated that using the better understanding of principal mechanisms of autoimmunity in the PNS will aid in the development of more specific and efficacious treatments. PMID- 8732199 TI - Immunology of the Schwann cell. AB - The Schwann cell or its myelin membrane appears to be the focus of autoimmune attack in several peripheral neuropathies. Potential Schwann cell antigens have therefore been extensively studied. Of the known Schwann cell proteins, MBP, P2, MAG, PLP, PO, PMP-22 and Connexin 32, gene defects of the latter three have recently been shown to be responsible for some forms of hereditary sensory and motor neuropathies. The L2/HNK-1 epitope, common to MAG, Po and PMP-22, is the target of autoantibody damage in neuropathies associated with certain IgM paraproteinaemias. P2, the inciting antigen in rat EAN, has no demonstrated role in human neuropathies. The same appears true for the glycolipid galactocerebroside, a neuritogen in rabbits. Gangliosides are currently under intense study, but the antigen for the common inflammatory neuropathies remains undefined. Evidence for the potential role of Schwann cells in immune modulation is provided; Schwann cells produced MHC molecules, the adhesion molecules I-CAM1, L1, L2-HNK-1, Ng-CAM, N-cadherisn, the cytokines IL-1, IL-6 and TNF-alpha and the inflammatory prostanoids, PGE-2 AND TxA2. PMID- 8732200 TI - Wallerian degeneration in peripheral nerve disease. AB - Wallerian-like degeneration can be regarded as the final shared pathway of the most prevalent diseases of the peripheral nervous system, including many degenerative, metabolic, heritable, toxic, inflammatory and ischaemic disorders. The cellular responses that take place during Wallerian degeneration, including the elaboration of neurotrophins, are increasingly recognized to set the stage for the success or failure of subsequent regeneration, and manipulations of Wallerian degeneration are being investigated as a potential means of altering the outcome of nerve regeneration. PMID- 8732201 TI - Animal models of neuropathies. AB - Neuropathy in animals is either genetically determined or is provoked by chemical compounds or physical injury. Diabetes in mice and rats may be spontaneous or induced, but a true copy of diabetic neuropathy in man is not yet available. Painful neuropathy occurs after nerve constriction or neuroma formation. A mouse mutant with delayed Wallerian degeneration demonstrates the pivotal role of this process for the regeneration of injured axons. Surprisingly, the neurotoxic effect of cisplatin which is severe in cancer patients has not yet unambiguously been reproduced in animals. Genetically determined diseases in mutants or transgenic animals may affect the myelination of peripheral axons. 'Trembler mice' are deficient in myelin and possibly correspond to CMT IA in man. The relation of sensory neuronopathies in mice, rats and dogs to human diseases is not yet clear. Motor neuronopathies in experimental animals have attracted much interest, because the recent discovery of motoneuronotrophic factors has raised high hopes. Most of the mutants described have not been appropriately studied, and the mouse mutant 'motoneurone disease' (mnd) eventually was found to have Batten's disease. None of the few more thoroughly studied models is probably a copy of human disease, although they may none the less help to test new therapies. PMID- 8732202 TI - The electrophysiology of axonal and demyelinating polyneuropathies. AB - Electrodiagnostic studies, which include nerve conduction studies, electromyography, repetitive nerve stimulation, single fibre EMG, autonomic function tests and quantitative sensory tests are a critical component of the neuromuscular evaluation. These studies, which are an extension of the clinical examination, help to classify further patients into the part of the peripheral nervous system affected and provide significant insight into the pathophysiological processes that underlie clinical illnesses. Traditionally, patients with peripheral neuropathy are classified as having demyelinating or axonal neuropathies. The methods of making this distinction are discussed. In addition, a number of additional studies are now available which provide further, insights into selected aspects of the peripheral nervous system. PMID- 8732203 TI - Magnetic stimulation in the assessment of peripheral nerve disorders. AB - Relatively pain-free excitation of both superficial and deep nerves in the assessment of nerve conduction velocity is the main advantage of magnetic stimulation over conventional electrical stimulation. General utility of this technique has often been called into question by a number of authors because of difficulties in obtaining supramaximal responses or in determining the exact site of impulse generation when stimulating a peripheral nerve distally. Meanwhile, magnetic stimulation of the cervical and lumbar roots has become a routine procedure for the assessment of peripheral conduction time and is combined with transcranial magnetic stimulation of the motor cortex in the assessment of central conduction time. Recent developments in magnetic coil and stimulator design have improved the focality of the stimulus, so that selective supramaximal stimuli can be delivered to commonly studied peripheral nerves in the upper and lower limbs, both at proximal and distal segments. Furthermore, the introduction of small figure-8-shaped coils enables safe diagnosis of chronic compression syndromes with exact assessment of conduction velocities over short distances of peripheral nerves. The ease of application of magnetic stimulation and the absence of pain for the patient make magnetic stimulation a particularly attractive method for also investigating patients with demyelinating polyneuropathies, and will certainly replace conventional electrical stimulation in the near future. PMID- 8732204 TI - Immunocytochemistry as a diagnostic tool. AB - ICC is a powerful technique to gain information about pathological processes in peripheral nerve beyond the limits of classical histopathology. ICC can aid in the identification of cellular infiltrates (T cells, B cells and macrophages), in the definition of their state of activation (MHC class I and II molecule expression), in the localization of cellular adhesion molecules (vascular and intercellular adhesion molecule 1) involved in trafficking of inflammatory cells through the endothelium and, finally, in the identification of locally produced cytokines (interferon gamma, interleukins and tumour necrosis factor). Moreover, ICC can identify pathological deposits, such as immunoglobulins and amyloid, within nerves and can be used to study the expression of myelin sheath proteins as they change in disease. In this chapter, basic immunocytochemical findings in experimental diseases of the peripheral nervous system, such as WD and immune mediated demyelination, are summarized and discussed in the context of similar observations in sural nerve biopsies. PMID- 8732205 TI - Electron microscopy as a tool to diagnose neuropathies. AB - Ultrastructural examination of a peripheral nerve biopsy may be particularly useful and sometimes indispensable for identification of the type of nerve lesion and of the aetiologies of peripheral neuropathies. The ultrastructural findings have anyway to be correlated with the clinical findings, the electrophysiological examination and the laboratory investigations. In this presentation, the various causes of peripheral neuropathies for which nerve biopsy study by electron microscope can provide diagnostic information are discussed. The principal aetiologies that will benefit from such an ultrastructural study are toxic, infectious, haemopathic, genetic and storage disorders. Sometimes the electron microscopic examination will help to determine not only the cause of the peripheral neuropathy, but also the mechanism of nerve lesions. Thus, in peripheral nerve pathology, ultrastructural study is an efficient tool among other available, complementary techniques. PMID- 8732206 TI - Undiagnosed neuropathies: the impact of ancillary investigations. AB - Before the advances in knowledge about peripheral nerve disorders that have accumulated over the past two to three decades, a high proportion of chronic neuropathies seen in tertiary referral centres remained undiagnosed. With intensive investigation, it is now possible to reduce this proportion to 13-14%. The category for which an aetiological diagnosis is most elusive is that of chronic axonal polyneuropathies, although some multifocal neuropathies resist explanation. An aetiological diagnosis is achieved for most demyelinating neuropathies. Some examples of chronic idiopathic axonal polyneuropathy (CIAP) may be of genetic origin; others may have an autoimmune basis. Advances in the molecular genetics and immunopathology of peripheral nerve disorders are likely to lead to the elucidation of some of those neuropathies that currently have to be classified as cryptogenic. PMID- 8732207 TI - Leprous neuropathies. AB - Once a terrifying disease, leprosy today has a very hopeful prognosis, provided that it is diagnosed early and treated with modern multidrug chemotherapy, any immunological reactions being recognized quickly and controlled well to prevent (further) peripheral nerve damage after commencing treatment. The diagnosis should be considered in all patients who present with peripheral neuropathy and/or anaesthetic skin lesions who come from or have lived in the tropics and subtropics. Although M. leprae cannot yet be grown in vitro, it is readily grown in experimental animals. A complete gene library has been developed, much of the genome mapped and a number species-specific and common mycobacterial antigens identified. The intricacies of the host-parasite relationship, especially of cell mediated immunity, and of the important immunological reactions of ENL and reversal reaction have been widely investigated. Modern MDT has caused a dramatic fall in prevalence, although the world annual case detection rate remains at around 600,000 new patients, many being at an early stage of the disease. WHO has launched a campaign to eliminate leprosy as a significant public health risk by the 2000 (with a prevalence of less than 1:10 000 population), which may well be achieved in some endemic countries. Leprosy will, however, remain an important cause of peripheral neuropathy for at least several more decades. PMID- 8732208 TI - Neuropathies in HIV infection. AB - Peripheral neuropathies represent the most common neurological manifestation in patients infected with HIV infection occurring either early in the infection or during the course of the illness. They present as acute or chronic demyelinating neuropathies (Guillain-Barre syndrome or chronic inflammatory demyelinating polyneuropathy), mononeuritis multiplex, ganglioneuronitis, cytomegalovirus related polyradiculoneuropathy, autonomic neuropathy or distal painful sensory neuropathy. They are multifactorial in aetiology. Their putative cause (viral, autoimmune, toxic, nutritional, co-infections) are often dictated by the stage of the underlying HIV disease. The virus, which is not found within ganglionic neurones or Schwann cells but only within the endoneurial macrophages, may generate a tissue-specific autoimmune attack by secretion of cytokines that promote trafficking of activated T cells and macrophages within the endoneurial parenchyma. The wide use of the neurotoxic antiretroviral nucleoside analogues ddC, ddI, d4T and 3TC, exacerbate or trigger subclinical neuropathy in many of these patients. PMID- 8732209 TI - Paraproteinaemic neuropathies. AB - Paraproteinaemia and neuropathy are each relatively frequent and may be associated by chance. However, a number of significant relationships have to be ruled out in the differential diagnosis. Malignant gammopathy should be excluded: multiple myeloma can lead to compression of the spinal cord or cauda equina; primary amyloidosis is occasionally involved; the rare but intriguing POEMS syndrome, consisting of polyneuropathy, organomegaly, endocrinopathy, M-protein and skin changes, usually accompanies osteosclerotic myeloma. It can be associated with angio-follicular lymph node hyperplasia and needs to be recognized because radioablative therapy is curative. The 'benign' monoclonal gammopathies of undetermined significance, known as MGUS, are much more frequent. There is an IgM MGUS group with predominantly distal sensorimotor demyelinating polyneuropathy and another rather heterogeneous group with IgG or IgA MGUS and a tendency to a favourable response to plasmapheresis. The role of the monoclonal IgG and IgA antibodies is unclear. This chapter has focused on the pathogenetic mechanisms of neuropathies associated with IgM MGUS. In the majority of cases, monoclonal autoantibodies specific for particular carbohydrate epitopes bind to myelin and are now recognized as the primary cause of the disease manifestations, including widening of the myelin lamellae. While the autoantibodies have been shown to bind complement, the presence of inhibitors is invoked to explain the absence of acute inflammatory changes. The epitopes recognized with the highest affinity by the auto-antibodies are present on the myelin-associated glycoprotein (MAG) and could interfere with cell adhesion and cellular signally processes. In addition, binding to antigenically similar glycoproteins, such as PO, PMP-22 and some acidic glycolipids, may be a contributory factor. It is generally accepted that the anti-MAG autoantibodies are inducing a progressive demyelinating polyneuropathy by modifying axon-Schwann cell interactions. PMID- 8732210 TI - Immune neuropathies in childhood. AB - In contradistinction to older populations, immune-mediated disorders (principally demyelinating processes) account for nearly half of peripheral neuropathies in childhood. The largest single diagnostic entity is GBS, which makes up approximately 25% of sensorimotor neuropathies in patients under 18 years of age. The clinical features are similar to those encountered in adults, although the prognosis in youngsters appears to be better than in older populations. Despite the absence of prospective data, plasmapheresis seems to be an effective modality for hastening recovery during GBS in children. The use of human immunoglobulin has gained acceptance for the treatment of GBS in adults, but insufficient data exist to draw firm conclusions about it role in the management of paediatric GBS. CIDP is the second most common cause of chronic sensorimotor neuropathy in children. The clinical manifestations of this disorder are extremely variable, and it can mimic the phenotype of several genetically determined neuropathies. The prognosis in this disorder is also relatively good, although a small number of children have significant neurological disability or treatment side-effects. Other immune-mediated neuropathies are relatively infrequent in our experience. When they occur, they are often associated with collagen-vascular diseases or bone marrow transplantation. Peripheral neuropathy in association with HIV infection in children appears to be rare. PMID- 8732211 TI - Ambulance or helicopter. PMID- 8732212 TI - Day-case procedures. PMID- 8732213 TI - Perianal and anorectal conditions. AB - Day-case surgery applies equally to colorectal surgery as any other discipline in surgery. Careful patient selection is the key to a successful outcome. All patients must be examined before the procedure as a change in symptoms and signs is likely to modify the choice of procedure. This review examines day-case management of common perianal and anorectal conditions. PMID- 8732214 TI - Gamete intrafallopian transfer. AB - Gamete intrafallopian transfer (GIFT) is a form of assisted reproduction which may be used as an alternative to in vitro fertilization (IVF) in certain circumstances. It is undertaken as a single day-case episode and requires less embryological expertise than IVF. GIFT has been successfully carried out in a number of district general hospitals. The details of the procedure as presently undertaken in Wigan are described. PMID- 8732215 TI - The training implications of day-case surgery. AB - The marked increase in the use of day surgery has major implications for the provision of surgical training. It is essential that day surgery units be used effectively for surgical training. We identify current constraints on such training and suggest methods for improvement. PMID- 8732216 TI - Recent advances in the aetiology and management of pre-eclampsia. AB - Despite advances in antenatal and intrapartum care, hypertensive disorders remain the second most common cause of maternal mortality. The Confidential Enquiry into Maternal Deaths in the United Kingdom (1988-90) (Department of Health, 1994) demonstrated that the number of deaths where hypertensive disease was the direct cause had not decreased since the previous trienium. The morbidity from this condition remains largely unquantified and many aspects of the aetiology and disease processes remain unknown. PMID- 8732217 TI - Pass the Royal College examinations. MCQ technique. AB - Trainee doctors can fail Royal College exams as a result of poor multiple choice question technique. On a negatively marked exam it is possible for candidate to know a subject well, answer 72% of the questions correctly, and yet still only obtain a mark of 44%. As a result, even some very good clinicians fail these exams. PMID- 8732218 TI - Hypotensive anaesthesia. AB - Hypotensive anaesthesia can be employed to improve the outcome in a variety of surgical procedures. Since there is no single agent capable of safely and effectively lowering arterial pressure in all situations, the anaesthetist may need to employ a variety of agents or techniques in order to achieve the target pressure. This article considers the currently available methods of inducing hypotension and their application to the safe conduct of anaesthesia. PMID- 8732219 TI - Urology image quiz. Erythroplasia of Queyrat. PMID- 8732220 TI - Nitric oxide: physiological roles and therapeutic implications in the lung. PMID- 8732221 TI - New technology in the diagnosis of tuberculosis. PMID- 8732222 TI - Gene therapy for cystic fibrosis: current issues. PMID- 8732223 TI - Predicting acute respiratory distress syndrome and intrapulmonary inflammation. PMID- 8732224 TI - The natural history of osteoporosis. AB - Osteoporosis remains a formidable challenge to medicine. A greater understanding of the variables that influence bone mineralization will allow the development of strategies to determine those at risk of developing osteoporosis. Advances in the management of this disease will probably come from prevention and selective screening. PMID- 8732225 TI - Management of diabetic ketoacidosis in childhood. AB - Diabetic ketoacidosis remains a life-threatening condition. Management is usually straightforward, but there is still a significant mortality and morbidity, largely arising from the unpredictable complication of cerebral oedema. The pathophysiology of this devastating condition is still unknown. Good supervision from senior members of staff is essential, and there should be early concern if progress is not as predicted. Rapid intervention with mannitol and hyperventilation is necessary if signs of cerebral oedema develop. PMID- 8732226 TI - Serum creatinine and urea: make the most of these simple test. AB - The most frequently requested measurements of renal function are serum urea and creatinine. This article describes how to make the most of these simple tests in the assessment of patients with renal disease. PMID- 8732227 TI - Neuroleptic malignant syndrome. AB - Neuroleptic malignant syndrome is a rare, idiosyncratic condition related to neuroleptic use which may develop at any stage during neuroleptic treatment and can prove fatal. Although most commonly seen in psychiatric patients, anyone prescribed a neuroleptic is at risk and it has been associated with other agents. This article discusses its recognition, risk factors, clinical course and treatment, together with a summary of current thinking on its aetiology. PMID- 8732228 TI - Phentolamine abolishes the pain of chronic pancreatitis. PMID- 8732229 TI - Nursing in the NHS: the executive nurse director. AB - The advent of trusts within the NHS brought a new dimension to the role of nurses in management, in that each trust must appoint an executive nurse director to sit on the trust board. The function and potential function of this role is, I believe, pivotal to the development of clinical services within the NHS. PMID- 8732230 TI - Curricular training in general surgery. PMID- 8732231 TI - Reflexly runny noses: neurogenic inflammation in the nasal mucosa. PMID- 8732232 TI - Histamine H1-receptors in the nasal mucosa: a mystery solved? PMID- 8732233 TI - Unilateral nasal allergen challenge leads to bilateral release of prostaglandin D2. AB - BACKGROUND: Multiple mediators including prostaglandin D2 and leukotriene B4 have been shown to increase in nasal secretions during the early response to nasal challenge with antigen. OBJECTIVE: Our objective was to investigate the time course of prostanoid and leukotriene B4 release into nasal secretions on both the ipsilateral and contralateral side after a unilateral nasal allergen challenge. METHODS: We performed a controlled, randomized trial. Six volunteers were challenged unilaterally with antigen or diluent in a randomized order and discs were used to collect nasal secretions from both nostrils at 2 min intervals for 20 min after the challenge. Prostanoids and leukotriene B4 (LTB4) in recovered nasal secretions were measured by combined capillary gas chromatography-negative ion chemical ionization mass spectrometry (GC/MS). RESULTS: Nasal allergen challenge resulted in a significant and immediate increase in symptoms and sneezing. PGD2 was significantly elevated above diluent values (0.6 +/- 0.6 pg) 30 s after removal of the allergen disc (P < 0.05), reached its peak (423.2 +/- 182.4 pg) at 2 min and then slowly decreased. PGD2 also increased on the contralateral side after unilateral allergen challenge, reaching peak values about six times lower than on the ipsilateral side (70.8 +/- 21.7 pg at 6 min). Levels of 9a, 11b-PGF2 after antigen provocation became significantly higher than after diluent (0 +/- 0 pg) on the ipsilateral side at 2 min (17.2 +/- 5.9 pg), and reached peak levels at 4 min (25.1 +/- 8.0 pg). LTB4 also increased significantly on the side of challenge. For the other prostanoids measured (PGF2, PGF2 alpha, TxB2, 6kPGF1 alpha), no significant changes in either ipsilateral or contralateral secretions were observed after allergen challenge. CONCLUSIONS: Our study described the kinetics of PGD2 and LTB4 release as well as the contralateral release of PGD2. PMID- 8732234 TI - Increased expression of histamine H1 receptor mRNA in allergic rhinitis. AB - BACKGROUND: Histamine plays an important role in producing nasal symptoms via histamine H1 receptor (H1R) in allergic rhinitis. It is reported that the minimum histamine concentration that induces sneezing is lower in allergic patients than in normal control subjects. Previous studies by binding assay on H1R gave divided results on whether the number of H1Rs is increased in allergic rhinitis or not. OBJECTIVE: The objective of this study was to examine if H1R mRNA expression is increased in patients with allergic rhinitis compared with normal healthy volunteers. METHODS: We extracted RNA from scrapings of inferior turbinate mucosa of 10 patients suffering from allergic rhinitis and 10 control subjects. As the H1R gene lacks introns, we treated RNA pellets by DNase to distinguish RNA from contaminating genomic DNA. Since amplification of H1R and beta-actin mRNA remained in an exponential phase at 35 cycles, H1R and beta-actin mRNAs were amplified for 35 cycles by reverse transcription-polymerase chain reaction (RT PCR). The PCR products were hybridized with internal probes and band intensities were quantitated by a densitometer. RESULTS: The mean +/- SD of H1R/beta-actin ratio was 0.88 +/- 0.62 for the patients with allergic rhinitis and 0.29 +/- 0.17 for the normal subjects; the difference was statistically significant (P < 0.01). CONCLUSIONS: These data suggest that expression of H1R mRNA is increased in the nasal mucosa of the patients with allergic rhinitis. PMID- 8732235 TI - The respiratory effects of reduction of mite allergen in the bedrooms of asthmatic children--a double-blind controlled trial. AB - BACKGROUND: Inhalation of house dust mite (HDM) allergen may provoke attacks of asthma. OBJECTIVE: We investigated whether a double-blind placebo-controlled community-based study aimed at reducing the HDM allergens in the bedrooms of HDM sensitive asthmatic children using the best methods available would prove beneficial to the children's health. METHODS: The children (mean age 9.9 years, 34 boys) were recruited by a questionnaire submitted to 7386 families in a geographically-defined area of the UK. Subjects were chosen to take part in the double-blind placebo-controlled trial if they were asthmatic, skin sensitive to mites, and had mite allergen in their mattresses. Seventy children were randomly allocated to groups. In the active group, the children's bedrooms were treated with an acaricide (Acarosan) and the mattresses, pillows and duvets were encased in exclusion covers. The control group received placebo treatments. RESULTS: Forty-nine complete data sets were obtained. Applying bedding covers and Acarosan led to a median reduction of 480 ng (100%) in mite allergen on the mattress vs 215 ng (53%) reduction in placebo-treated group by 6 weeks. No evidence was found that the acaricide reduced mite allergen level. A change in bronchial reactivity to histamine was observed in the children after 6 weeks. This was not associated with any change in thrice-daily records of peak expiratory flow rate. By 24 weeks, the actively-treated children had improved forced expiratory volume in 1s (FEV1) and fewer required bronchodilator therapy or reported asthmatic symptoms than did the controls. CONCLUSIONS: The results suggest that mite removal procedures may modestly improve mite-sensitive asthmatics and could perhaps be of value in exceptionally mite-sensitive and/or highly mite-exposed individuals whose response to the attempted removal should be measured. PMID- 8732236 TI - Decreased production of IFN gamma and increased production of IL-6 by cord blood mononuclear cells of newborns with a high risk of allergy. AB - BACKGROUND: The underlying mechanisms of elevated IgE level in atopic patients are still obscure, however, extensive efforts have been tried to identify an immunological parameter as a predictor of atopy. OBJECTIVE: This study compared the difference in cytokine production by cord blood mononuclear cells between new borns with high-risk of allergy (family allergy score, FAS > or = 3) and those with low-risk (FAS = 0). METHODS: Following stimulation with PHA (100 micrograms/mL) and PMA (1 ng/mL), the cytokines produced by cord blood CD4+ T cells in the presence of monocytes were measured by ELISA kits and the mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR) technique. RESULTS: Our results showed: CD4+ T cells in the presence of monocytes and isolated monocytes from the high-risk group produced a much greater amount of IL 6, either spontaneously or after stimulation, than did those of the low-risk group; CD4+ T cells of low-risk group produced a significantly greater amount of interferon gamma (IFN gamma) than did those from the high-risk group; IL-4 cannot be detected by ELISA kit, and only a trace amount of IL-4 mRNA was detected by RT PCR technique; cord blood basophils stimulated with PHA and PMA could produce a significant amount of IL-4; there was an inverse correlation between the production of IFN gamma and cord blood IgE level (high-risk group, r = 0.647, n = 17) and the number of natural killer (NK) cells (CD3- CD16+ CD56+) was significantly lower in high-risk group than for low-risk group. CONCLUSION: Our data suggested increased production of IL-6 and decreased production of IFN gamma of cord blood mononuclear cells appear to be the hallmark of newborns from the high-risk population. PMID- 8732237 TI - Allergenic and antigenic determinants of latex allergen Hev b 1: peptide mapping of epitopes recognized by human, murine and rabbit antibodies. AB - BACKGROUND: The rubber elongation factor in Hevea rubber (Hev b 1) is one of the most important latex allergen and is leading cause of latex type 1 hypersensitivity in children with spina bifida. OBJECTIVE: The aim of this study was to define the allergenic and antigenic epitopes of Hev b 1. METHODS: The immunoglobulin- (Ig)E and IgG antibody binding sites on Hev b 1 allergen were delineated by enzyme linked immunosorbent assay (ELISA) using synthetic overlapping peptides covering the whole Hev b 1 sequence. In order to improve the binding capacity and specificity all peptides were biotinylated at the N-terminal end via a 6-aminohexanoic acid as spacer and then adsorbed to streptavidin pre coated microtitre plates. Fine mapping to define the essential amino acid residues for the antibody binding was achieved by using overlapping peptides with one amino acid offset. RESULTS: It was demonstrated that the IgE epitopes were located in different regions of Hev b 1 including the C-terminal segment (121 137) and the segments with amino acid residues of 30-49 and 46-64. Two monoclonal antibodies (MoAbs) II2F3 and II4G9 raised against purified Hev b 1 recognized the C-terminal segment only. The results of epitope mapping with three rabbit antisera revealed that five positive peptides, including the epitope peptides 31 49, 46-64 and 121-137, were involved in the antibody-binding sites. Fine mapping on the segments 46-64 and 121-137 showed that the two MoAbs reacted with the peptide 125-134 in the C-terminal region, whereas the peptide with amino acids 124-134 was essential for recognition by human IgE antibodies. Epitopes to rabbit polyclonal IgG and human IgE were also found to be involved in the amino acid residues of 47-59. CONCLUSION: Our results indicate that the most allergenic/antigenic portions of Hev b 1 allergen are the C-terminal region and the region with amino acid residues of 31-64. In both regions, the minimal IgE binding epitope is almost identical with the IgG-binding epitope. PMID- 8732238 TI - Latex allergy can induce clinical reactions to specific foods. AB - OBJECTIVE: The purpose of this study was to investigate crossreactivity between latex and foods, to identify crossreacting IgE binding proteins, and to assess the clinical significance. METHODS: Forty-seven latex allergic patients and 46 non-latex allergic patient controls were studied. Allergen sensitization was determined by skin-prick testing (SPT) and allergenic proteins were identified by immunoblot reactivity and amino acid sequence analysis. RESULTS: Immunological reactivity to foods was found to be common, occurring in 33 latex-allergic individuals but in only seven controls (P < 0.000001); 100 of 376 (27%) food skin prick tests were positive in the latex-allergic subjects. Twenty-seven out of 100 positive food SPTs were associated with clinical symptoms. Seventeen patients manifested a clinical allergy to at least one food including 11 with anaphylaxis, and 14 with local sensitivity reactions. Positive food skin tests occurred most frequently with avocado (53%), potato (40%), banana (38%), tomato (28%), chestnut (28%), and kiwi (17%). Latex-allergic patients (23%) recognize a protein that had sequence homology to a broad class of plant proteins known as patatins. Crossreactivity between latex and several potato proteins was observed by immunoblot inhibition analysis. CONCLUSIONS: Sensitization to latex has extensive crossreactivity with certain foods and leads to clinical allergic reactions. Potatoes and tomatoes are newly reported cross-reacting foods. Plant proteins with structural homology to latex proteins may predispose to food allergy. PMID- 8732239 TI - Buckwheat flour hypersensitivity: an occupational asthma in a noodle maker. AB - BACKGROUND: There have been very few reports of occupational allergies caused by inhalation of buckwheat flour. In this paper, we present a case of occupational asthma and rhinitis caused by buckwheat flour inhalation. METHODS AND RESULTS: The patient had strong positive responses to grass and ragweed pollens as well. The bronchoprovocation test showed early asthmatic response to buckwheat flour extracts. Serum specific IgE antibody to buckwheat flour was detected by enzyme linked immunosorbent assay (ELISA). In order to further identify the allergenic component of the extracts, sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and electroblotting studies were performed. Eight IgE binding components (9-55 kDa) were detected within the buckwheat flour extracts. CONCLUSION: These results suggest that inhalation of buckwheat flour can caused IgE mediated bronchoconstriction. PMID- 8732240 TI - Rye flour allergens associated with baker's asthma. Correlation between in vivo and in vitro activities and comparison with their wheat and barley homologues. AB - BACKGROUND: A number of wheat and barley flour proteins that belong to the cereal alpha-amylase/trypsin inhibitor family have been identified as major allergens associated with baker's asthma. However, the allergenic role of this protein family had not been investigated in rye. OBJECTIVE: To study the allergenicity of five purified proteins from rye flour which belong to the same inhibitor family, as well as to compare their properties with those of their wheat and barley homologues. METHODS: In vivo skin-prick tests were carried out in 21 patients with radioallergosorbent test (RAST) 2-3 to rye and allergic sensitization mainly to this cereal flour. In addition, sera from all these patients were used to assay the IgE binding capacity of dot blotted purified proteins. RESULTS: Three of the rye proteins tested, namely Sec c 1, RDAI-1 and RDAI-3, provoked positive skin-prick tests in more than 50% of patients, although their in vitro reactivity was lower. Different reactivities were found for the rye components compared with their wheat and barley homologues. Statistical analyses showed a significant correlation between the results of in vivo and in vitro tests for seven out of the nine purified proteins considered in this study. CONCLUSION: Members of the rye alpha-amylase inhibitor family are main allergens involved in allergic reactions to cereal flours. However, different allergenic behaviours were found between homologous allergens from rye, barley, and wheat. PMID- 8732241 TI - Stability of cereal allergens. AB - BACKGROUND: Ingested cereals and inhaled cereal dusts can cause IgE-mediated allergy. Testing for these allergies yields variable, often undependable results, which could be due to the use of unsuitable or degraded testing materials. OBJECTIVE: We studied the effect of storage media, time and temperature on the stability of protein allergens extracted from wheat, rye, barley and oats flour. We also examined the effect of heat processing similar to that used when preparing foods on the stability of allergenic proteins present in wheat flour. METHODS: After storage experiments proteins in allergen extracts were separated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and then visualized using immunoblotting or staining with Coomassie brilliant blue. After heat processing wheat flour proteins were extracted and their IgE-binding capacity was estimated. RESULTS: Extracted proteins from wheat, rye and barley flour appeared stable for at least 21 months when stored at 4 degrees C in an aqueous solution containing 50% glycerol and 0.45% sodium chloride. In a medium with 0.4% phenol and 0.9% sodium chloride degradation of many proteins became apparent already after 1 month of storage. Oats extracts lost their IgE-binding capacity more rapidly and the maximal storage time appeared to be 10 months at 4 degrees C in a solution containing 50% glycerol and 0.45% sodium chloride. The IgE-binding capacity of the allergenic proteins decreased as heat-processing temperature and heating time increased. No processing conditions entirely abolished IgE-binding. CONCLUSION: Cereal allergen preparations can be stored either in lyophilized form or at 4 degrees C in 50% glycerol. Unheated materials should be used also when testing for food allergy, as heating reduces allergenicity and thus sensitivity at testing. PMID- 8732242 TI - Allergenic components in three different species of Penicillium: crossreactivity among major allergens. AB - BACKGROUND: Penicillium species have been considered as important causative agents of extrinsic bronchial asthma. However, little is known about the allergens of these ubiquitous airborne fungal species. OBJECTIVE: This study compares the allergenic profiles and allergenic crossreactivity among allergens of three prevalent airborne Penicillium species. METHODS: IgE-binding Penicillium components were identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)-immunoblotting using sera from 67 asthmatic patients. The presence of allergenic crossreactivity was analysed by immunoblot inhibition. RESULTS: Among the 67 serum samples tested, 15, 14 and 11 samples showed IgE reactivity to components of P. citrinum, P. notatum and P. brevicompactum, respectively. All 15 P. citrinum-positive serum samples showed IgE-binding to a 33 kDa extract component of this species. Thirteen (93%) of the 14 P. notatum positive serum samples and 10 (91%) of the 11 P. brevicompactum-positive sera also showed IgE reactivity to components with a molecular weight of about 33 kDa in individual Penicillium species. All of the 10 P. brevicompactum 33 kDa component-positive serum samples showed IgE reactivity to the 33 kDa components of the other two Penicillium species tested. Dose-dependent inhibition of IgE binding to these major allergens was observed when the positive serum sample was absorbed with different amounts of individual allergenic extract as well as with different amounts of extracts prepared from the other two Penicillium species. CONCLUSION: Although different allergenic profiles were observed in the three different Penicillium species tested, results showed that there was an IgE crossreactivity among the 33 kDa group major allergens of P. citrinum, P. notatum and P. brevicompactum. PMID- 8732243 TI - Enhanced IgE response to Candida albicans in postoperative invasive candidiasis. AB - BACKGROUND: Invasive candidiasis is a life-threatening complication problem in post-operative and immunocompromized patients, e.g. those treated by intensive care. Candida is frequently cultured from the mucous membranes of hospital patients and fungal cultures offer little diagnostic help. Other diagnostic methods, such as blood cultures, serology and diagnostic imaging techniques produce results too late and, if positive, low sensitivity. OBJECTIVE: To study the value of Candida-specific antibodies, especially those of IgE class, in diagnosing invasive Candida infection. METHODS: The immunoglobulins IgE, IgG and IgM responses to antigens of Candida albicans in the sera of 14 patients with culture, biopsy and/or autopsy proven postoperative invasive candidiasis and of 11 colonized and 19 non-colonized operated patients were studied by mannan radioallergosorbent test (RAST), mannan enzyme-linked immunosorbent assay (ELISA) and immunoblotting. RESULTS: Detection of IgE antibodies to C. albicans polysaccharide (mannan) and protein antigens proved specific and sensitive in diagnostics of invasive candidiasis after major abdominal surgery. IgE rose early in the course of the infection and the method made a clear distinction between invasive infection and mucous colonization. Immunoblotting for protein antibodies was most sensitive while nitrocellulose-RAST for mannan antibodies was most specific. The combined use of immunoblotting and RAST increased the sensitivity and the specificity. Determinations of anti-Candida IgG and IgM antibodies had low sensitivity and specificity. CONCLUSION: Critically ill patients with invasive candidiasis develop IgE antibodies to Candida antigens probably because of disturbed TH1/TH2 responses. Determination of specific IgE antibodies can be used as a diagnostic aid in the early stage of invasive Candida infection. PMID- 8732244 TI - Immunohistochemical detection of GM-CSF, IL-4 and IL-5 in a murine model of allergic bronchopulmonary aspergillosis. AB - BACKGROUND: Granulocyte macrophage-colony stimulating factor (GM-CSF), interleukin-4 (IL-4) and IL-5 are important in tissue eosinophil accumulation and high IgE production in allergic inflammatory reaction. OBJECTIVE: We examine lung GM-CSF, IL-4 and IL-5 expression in a murine model of allergic bronchopulmonary aspergillosis (ABPA) characterized by eosinophil and lymphocyte lung infiltration and elevated serum IgE level. METHODS: C57BL/6 mice were intranasally treated three times a week for 1, 2 or 3 week(s) with Aspergillus fumigatus (Af) antigen or saline and were sacrificed on days 7, 14 and 21. Immunohistochemical analyses for GM-CSF, IL-4 and IL-5 were performed on lung sections. RESULTS: Af treatment induced a remarkable pulmonary eosinophil influx. Increased numbers of lung T lymphocytes and GM-CSF positive cells were observed on days 14 and 21. IL-4 and IL-5 positive cells were increased significantly only on day 14. Immunostained serial sections showed that most (> or = 98%) cytokine positive cells were CD3 positive. Few eosinophils (< 2% of cytokine positive cells) were immunoreactive for GM-CSF and IL-5. Significant correlations were found between the number of GM CSF and IL-5 positive cells, and the number of eosinophils in Af-treated lung (r = 0.62, P < 0.05 and r = 0.52, P < 0.05 respectively), and between the number of IL-4 positive cells and the serum total IgE level (r = 0.64, P < 0.01). CONCLUSIONS: Our data suggest a role for T lymphocyte GM-CSF, IL-4 and IL-5 in Af induced mouse pulmonary eosinophilia and increased serum IgE production and further support the importance of T helper (TH2) cells in the pathogenesis of ABPA. PMID- 8732245 TI - Influence of endogenous cortisol on eosinophil function in sensitized pigs: direct measurements of eosinophil peroxidase. AB - BACKGROUND AND OBJECTIVES: Eosinophils are associated with bronchial asthma, but the role of the eosinophil is not fully understood. This study was initiated in order to study the influence of endogenous cortisol on eosinophil recruitment and activation in allergic inflammation in the lower airways in the pig. METHODS: Polyclonal and monoclonal antibodies against porcine eosinophil peroxidase (EPO) were raised. Detection of eosinophils in blood smears and lung biopsy specimens was achieved using the polyclonal antibody. For determination of porcine EPO in bronchoalveolar lavage (BAL) fluid, a sandwich enzyme-linked immunosorbent assay (ELISA) with a detection limit of 0.15 micrograms/L was developed. No cross reactivity with porcine myeloperoxidase was found. Pigs that had been actively sensitized with repeated subcutaneous injections of Ascaris suum antigen were acutely challenged with antigen in the lower airways under pentobarbitone anaesthesia. RESULTS: Control animals with plasma cortisol levels of approximately 400 nM did not exhibit infiltration of eosinophils into lung parenchyma or EPO-release in the bronchial lumen within 8 h after challenge. However, in pigs treated with a cortisol-synthesis inhibitor (metyrapone), resulting in plasma cortisol levels of approximately 40 nM, there was a marked eosinophil infiltration into lung tissue at 8 h. Furthermore, EPO levels in BAL fluid were increased in some, although not all, low-cortisol animals. There was no infiltration of eosinophils into skin tissue in these animals. CONCLUSIONS: It is concluded that, after allergen challenge in the lower airways of metyrapone treated pigs, newly recruited eosinophils infiltrate lung tissue specifically. Furthermore, a cortisol-sensitive release of the eosinophil-derived cationic protein EPO, into the bronchial lumen was established. This is, to our knowledge, the first description of direct measurements of the release of an eosinophil granule protein in a large animal model of allergy. PMID- 8732246 TI - Restoration of cardiac contraction by angiotensin II during raised [K+]O in the rabbit. AB - Catecholamines restore cardiac contraction depressed by hyperkalaemia (raised [K+]O) and acidosis, yet in exercise hyperkalaemia and acidosis are tolerated during beta adrenergic blockade. To test whether the negative effects of raised [K+]O are offset by a non-adrenergic hormone, angiotensin II (AII) was given to rabbit papillary muscle (All 75 nM, n = 9) and rabbit isolated working hearts (All 5 nM, n = 8) perfused with 8 and 10 mM K+ Tyrode at 37 degrees C. A similar protocol was also performed in a further nine isolated hearts treated with propranolol (1 microM) and prazosin (1 microM). All caused a significant (P < 0.01) increases in contraction and aortic flow in normal Tyrode and maintained aortic flow during high [K+]O. In the papillary muscle and isolated heart treated with adrenergic blockers, high [K+]O reduced the stimulatory effects of All, but contraction and aortic flow was still significantly greater (P < 0.01) than in high [K+]O alone. These results show that All can ameliorate the depressive effects of high [K+]O on the heart. The local release of All in the heart during activation of the sympathetic nervous system and the rise in circulating All during exercise could therefore play a role in protecting the heart from hyperkalaemia. PMID- 8732247 TI - Central command increases cardiac output during static exercise in humans. AB - Neural control of the circulation during static two-leg exercise was evaluated in 10 subjects. External compression of the legs was employed to assess muscle mechano-receptor influence by achieving the same intramuscular pressure (80 mmHg) as developed during exercise. The muscle metabo-reflex contribution was assessed by post-exercise muscle ischaemia, and the influence from higher centres in the central nervous system ('central command') was taken as the part of the response that could not be accounted for by the two reflex contributions. During static exercise, mean arterial pressure was higher (26 +/- 3 mmHg; P < 0.01) as compared with leg compression (10 +/- 2 mmHg) and with post-exercise muscle ischaemia (11 +/- 2 mmHg). Heart rate (25 +/- 4 b.p.m.) and cardiac output (0.8 +/- 0.3 L min 1) were increased only during static exercise (P < 0.05). Increase in total peripheral resistance were similar during static exercise, post-exercise muscle ischaemia and leg compression. The pressor response to static exercise with a large muscle group was equally attributable to mechanical and metabolic stimulation of afferent nerves; and the two influences were redundant in their effect on total peripheral resistance. In contrast, the influence from central command was directed to the heart with elevation of its rate and minute volume. PMID- 8732248 TI - Effects of preceding movements and contractions on the tonic vibration reflex of human finger extensor muscles. AB - The vibration sensitivity of feline muscle spindle endings is known to vary as a result of preceding muscle conditioning manoeuvres. If similar after-effects occur in man they should be expected to influence the strength of the tonic vibration reflex (TVR). To study this issue, vibration was applied over the finger extensor tendons of 11 volunteers who actively held their fingers in a semi-extended position. The TVR, measured as electromyographic responses and angular deflections at the metacarpophalangeal joints, was found to be stronger when the extensors prior to the test had been contracted in a shortened position than when they had been contracted in a stretched position. This difference was reduced when the vibration stimulus was preceded by a strong extensor contraction in the test position. The antigravity extensor EMG activity required to keep the fingers in the test position was weaker when the conditioning contraction was performed at a short muscle length than when it was performed at a long muscle length. The variations in magnitude of the TVR can be attributed to 'thixotropic' properties of intrafusal muscle fibres, resulting in a slack following conditioning at a long muscle length. The different EMG levels required for position holding can be explained by similar properties of extrafusal muscle fibres. In conclusion, the results demonstrate the importance of taking the 'history of movement' into account in the design of all TVR studies. PMID- 8732249 TI - Reversible depression of action potentials and force production in frog single muscle fibres by calmodulin-inhibitors. AB - The effects of the calmodulin-inhibitors trifluoperazine, thioridazine and zaldaride maleate on the responses to electrical stimulation in isolated frog skeletal muscle fibres were investigated. All three drugs initially reduced the amplitude of the action potentials but potentiated twitch force. This was followed by a total loss of action potentials and force production. However, the resting membrane potential was not changed. The effects were completely reversible upon removal of the drugs. These results suggest that an intact calmodulin system is required for normal function of the sarcolemmal sodium channels of frog skeletal muscle. PMID- 8732250 TI - Effects of perchlorate on myofibrillar calcium sensitivity in rat skinned skeletal muscles. AB - The effects of perchlorate (1-20 mM) on myofibrillar calcium responsiveness have been tested in Triton X-100-skinned fibre bundles from rat soleus (slow-twitch) and extensor digitorum longus (fast-twitch) skeletal muscles. In extensor digitorum longus and soleus, perchlorate dose-dependently shifted the pCa ( log[Ca2+])/tension relationship towards lower free calcium concentration (sensitizing effect) and maximal tension was unchanged. The degree of sensitization was greater in extensor digitorum longus than in soleus bundles. Reversibility after exposure to 12 mM perchlorate was complete in soleus but not in extensor digitorum longus muscles. In fact, the 'return' pCa/tension relationship in extensor digitorum longus was shifted to higher free calcium concentration (desensitizing effect) compared with control. Perchlorate (12 mM) also enhanced myofibrillar calcium responsiveness of frog semitendinosus skinned skeletal fibres. Assuming a passive distribution of perchlorate across the sarcolemma, this sensitizing effect is probably not involved in perchlorate induced potentiation of contractile responses of intact muscles and thereby supports the specificity of perchlorate as an agonist of the excitation/calcium release sequence in skeletal muscle fibres. PMID- 8732251 TI - Effects of strength and endurance training on isometric muscle strength and walking speed in elderly women. AB - The separate effects of 18 weeks of intensive strength and endurance training on isometric knee extension (KE) and flexion (KF) strength and walking speed were studied in 76- to 78- year-old women. Maximal voluntary isometric force for both KE and KF was measured in a sitting position on a custom-made dynamometer chair at a knee angle of 60 degrees from full extension. Maximal walking speed was measured over a distance of 10 m. The endurance-trained women increased KE torque and KE torque/body mass after the first 9 weeks of training when compared with the controls. when comparing the baseline, 9 week and 18 week measurements within the groups separately, both the endurance- and strength-training groups increased KE torque, KE torque/body mass and walking speed. Individual changes in KE torque/body mass before and after 18 weeks of training averaged 19.1% in the strength group, 30.9% in the endurance group and 2.0% in the controls. This study indicates that in elderly women the effects of physical training on muscle strength and walking speed occur after endurance as well as strength training. The considerable interindividual variation in change of muscle performance is also worth noticing. PMID- 8732252 TI - The effect of lesions of the sympathoadrenal system on training induced adaptations in adipocytes and pancreatic islets in rats. AB - Physical training increases insulin stimulated glucose uptake in adipocytes and decreases insulin secretion from pancreatic islets. The mechanism behind these adaptations is not known. Because in acute exercise adrenergic activity influences both adipocytes and pancreatic islets, the sympathetic nervous system was examined as the possible mediator. Rats were either adrenodemedullated or sham adrenodemedullated and underwent either unilateral abdominal sympathectomy or were sham sympathectomized. Resting plasma adrenaline concentration in adrenodemedullated rats was 32% of the concentration in sham adrenodemedullated rats (P < 0.0001) and muscle noradrenaline content in sympathectomized leg was 9% of content in sham sympathectomized leg (P < 0.0001). After operations rats were either swim trained for 10 weeks or remained sedentary. Insulin stimulated 3-O [14C]methylglucose transport was measured in adipocytes from epididymal fat pads, and insulin secretion and glucose metabolism were measured in glucose stimulated pancreatic islets. Training increased insulin stimulated glucose transport in adipocytes (P < 0.0001) and decreased their size (P < 0.0001), but neither adrenodemedullation nor sympathetic denervation affected these parameters significantly. Training decreased insulin secretion (P < 0.01) and increased glucose oxidation (P = 0.02) and utilization (P = 0.08) in pancreatic islets, but none of these parameters was affected significantly by adrenodemedullation. It is concluded that adrenergic activity is not important for the training induced decrease in size and increase in insulin stimulated glucose transport of adipocytes. Neither is an intact adrenal medulla necessary for training-induced adaptations in pancreatic beta cell function. Finally, in response to training, beta cell insulin secretion and glucose metabolism changed in opposite directions. PMID- 8732253 TI - The influence of extracellular Ca2+ on the time course of isovolumetric pressure development in the guinea-pig urinary bladder. AB - Isovolumetric contractions were induced electrically in five guinea-pig bladders in a 1.8 mM Ca2+ buffer solution at two different volumes. The rate of pressure development was studied by analysis of phase-plots, which are plots of the rate of change of intravesical pressure as a function of pressure. At relatively large volumes the rate of pressure development showed a biphasic nature, suggesting that two different processes were involved in different phases of contraction. Extracellular Ca2+ was manipulated to identify these processes. Removal of extracellular Ca2+ led to an 85% reduction of the maximum isovolumetric pressure in subsequent stimulations, but pressure developed faster. The biphasic nature of pressure development at large volumes tended to disappear. It was concluded that at large bladder volumes pressure development in this type of smooth muscle is determined by the release of intracellularly stored Ca2+ in the early phase of contraction. In the final phase the rate of pressure development is limited by the extracellular Ca2+ influx. At small bladder volumes the Ca2+ influx takes place at a higher rate, so that the contribution of intracellular Ca2+ release to pressure development could not be identified at these volumes. PMID- 8732254 TI - Shortening induced deactivation in the guinea-pig urinary bladder. AB - Shortening induced deactivation, the depressant effect of active muscle shortening on the subsequently measured isometric force, has been shown in smooth muscle strips and rings. The guinea-pig bladder permits the investigation of this phenomenon in a whole organ preparation. Previous work in our laboratory showed that shortening of the in vitro guinea-pig detrusor muscle had a depressant effect on the isovolumetric pressure that could be generated immediately afterwards. To test the hypothesis that this was caused by deactivation, the effects of active and passive detrusor shortening on the subsequently measured isovolumetric pressure were compared. The isovolumetric pressures measured after 5 min periods of recovery were taken as control values. It was found that the isovolumetric pressure after passive shortening was 7% smaller than the isovolumetric pressure without preceding shortening. This difference was ascribed to viscoelastic relaxation during shortening. Active shortening had an additional 8% depressant effect on isovolumetric pressure compared with passive shortening. The effects of active and passive shortening differed significantly. It was concluded that shortening induced deactivation in the guinea-pig urinary bladder smooth muscle in toto can be considered proven. The fact that deactivation is shown both by striated and smooth muscle preparations is in line with the assumption that it is caused by reduced actin-myosin interaction. The hypothesis that (in striated muscle) the latter is effected by a decrease in troponin calcium binding, however, needs reconsideration. PMID- 8732255 TI - Tachykinins increase vascular permeability in the gastrointestinal tract of the rat. AB - The occurrence of inflammation as indicated by extravasation of Evans blue bound to plasma proteins was examined in various parts of the gastrointestinal tract in the rat, following administration of tachykinins, capsaicin and hydrochloric acid. Intravenous neurokinin A dose-dependently induced extravasation in stomach, duodenum, jejunum, caecum and colon, but had no effect in ileum. Neurokinin B equipotently induced extravasation in the stomach but had no effect in other parts of the gut and substance P had no effects on extravasation of Evans blue in any of the examined parts of the gastrointestinal tract. Capsaicin given intraperitoneally increased vascular permeability in stomach and duodenum only, while extravasation of Evans blue after capsaicin given intraluminally did not differ from the effect of the vehicle alone. As a comparison, HCl given intraluminally in the duodenum was found to induce a prominent extravasation of Evans blue of a greater magnitude than than of tachykinins. We suggest that tachykinins, and in particular neurokinin A, may be of importance for extravasation of plasma proteins as part of inflammatory reactions in the upper and lower gastrointestinal tract. PMID- 8732256 TI - Extracellular distribution of insulin in muscle of rats exposed to long-term hyperinsulinaemia. AB - The importance of increased capillary density for the regulation of insulin sensitivity by transcapillary delivery of insulin to muscle cells in insulin exposed rats was investigated by direct microdialysis measurements of interstitial (125I] insulin concentrations in the femoral muscle during an euglycaemic hyperinsulinaemic clamp. In insulin-exposed rats plasma insulin was approximately 25% (P < 0.05) higher than that in control animals during the first 100 min and reached their maximal concentrations after 100 min. After a nitroprusside infusion given at 100 min both groups had similar concentrations of insulin in plasma as well as in muscle interstitial fluid. However, mean glucose infusion rate during the first clamp hour was 20.5 +/- 2.3 and 12.6 +/- 5.2 mg kg 1 min-1 (P < 0.05) in insulin-exposed and control animals, respectively. During the second clamp hour the corresponding figures were 21.1 +/- 2.4 and 13.9 +/- 2.6 (P < 0.05). It may be concluded that capillarization and/or nitroprusside affected plasma insulin concentrations without altering either the interstitial insulin levels or the insulin effect on glucose consumption. The data suggest that the elevated insulin sensitivity after chronic insulin exposure is dependent on other than transcapillary transport events and demonstrate the different kinetics for insulin distribution in plasma and in the interstitial fluid. PMID- 8732257 TI - Oleic acid binding and transport capacity of human red cell membrane. AB - Resealed human red cell membranes, ghosts, bind oleate (OL) by a limited number of sites when equilibrated at 37 degrees C, pH 7.3 with OL bound to bovine serum albumin (BSA) in molar ratios below 1.5. The binding capacity is 34 +/- 2.2 nmol g-1 ghosts with a dissociation equilibrium constant (37 degrees C) Kdm 1.38 +/- 0.15 fold Kd of albumin binding Kdm is temperature independent and approximately 7-8 nM. Exchange efflux kinetics at 0 degrees C to buffers of various albumin concentrations ([BSAy]) is biexponential and is analysed in terms of a three compartment model. Accordingly the ratio of inner to outer membrane leaflet binding sites is 0.450 +/- 0.018 and the rate constant of unidirectional flux from inside to outside is 0.067 +/- 0.01 s-1. The rate constant of flux from the extracellular side of the membrane to BSAy increases with the square root of [BSAy] as expected of an unstirred layer effect. This provides an estimate of the dissociation rate constant of OL-BSA complex at 0 degrees C of 0.0063 +/- 0.0003 s-1. Exchange efflux from ghosts containing four different [BSAi] obeys the expected kinetics of a three-compartment approximation of the theoretical model. Accounting for the effect of an unstirred fluid inside ghosts, the rate coefficients fit the values predicted by the parameters obtained by the studies of albumin-free ghosts. The results show that the OL transport across the membrane is mediated exclusively by the asymmetrically distributed binding sites. The differences between transport sites of three long-chain fatty acids suggest that they are protein determined microdomains of phospholipids. PMID- 8732258 TI - Millisecond studies of calcium-dependent exocytosis in pituitary melanotrophs: comparison of the photolabile calcium chelators nitrophenyl-EGTA and DM nitrophen. AB - DM-nitrophen (DMN) is a photolabile calcium chelator that has been used extensively to study calcium-triggered exocytosis. Nitrophenyl-EGTA (NPE) is a recently synthesized photolabile calcium chelator that, unlike DMN, selectively binds calcium over magnesium. Here, we compare NPE and DMN for their effectiveness in raising cytosolic calcium ([Ca]i) to trigger exocytosis. The whole cell patch clamp technique was used to monitor membrane capacitance (Cm) and to load both calcium indicator dye and photolabile chelators into rat pituitary melanotrophs prior to flash photolysis. In cells dialysed with DMN, a transient increase in [Ca]i was observed immediately after continuity between the patch pipette and the cell cytosol was achieved. This 'loading transient' reflects the release of calcium from DMN during the binding of intracellular magnesium. No such transient was seen with NPE, consistent with the negligible binding of magnesium to this chelator. Following flash photolysis of DMN or NPE, [Ca]i increased, triggering both a rapid exocytic burst and slower sustained phases of exocytosis. When flashes of the same intensity were compared, the photolysis of NPE resulted in smaller increases in [Ca]i and slower exocytic bursts than that of DMN. These findings are in accordance with the properties of the two compounds [Ellis-Davies G.C.R., Kaplan J.H. Nitrophenyl-EGTA, a photolabile chelator that selectively binds Ca2+ with high affinity and releases it rapidly upon photolysis. Proc Natl Acad Sci USA 1994; 91: 187-191] and the calcium dependency of the exocytic burst [Thomas P., Wong J.G., Lee A.K., Almers W. A low affinity Ca2+ receptor controls the final steps in peptide secretion from pituitary melanotrophs. Neuron 1993; 11: 93-104]. Although NPE is somewhat less effective than DMN in raising [Ca]i, this chelator promises to be a useful and interesting tool for the time-resolved study of calcium-dependent exocytosis in the presence of physiological concentrations of magnesium. PMID- 8732259 TI - 24,25(OH)2 vitamin D3 modulates the L-type Ca2+ channel current in UMR 106 cells: involvement of protein kinase A and protein kinase C. AB - In this study, the effect of 24,25(OH)2 vitamin D3 (24,25D3), on the L-type Ca2+ channel current (L-channel current) in UMR 106 cells was investigated using the whole cell version of the patch clamp technique. It was found that 24,25D3 had a dual effect on the L-channel current: a low concentration of 24,25D3 (1 x 10(-8) M) increased the amplitude of the L-channel current by 49 +/- 11%, whereas a high concentration of 24,25D3 (1 x 10(-5) M) reduced the amplitude of the current by 55 +/- 7%. The effect of a low concentration of 24,25D3 was mimicked by 8-bromo cAMP and inhibited by Rp-cAMPs, indicating the involvement of the cAMP/protein kinase A pathway. In contrast, the effect of a high concentration of 24,25D3 was mimicked by 4 beta-phorbol 12-myristate 13-acetate and inhibited by calphostin C, indicating the involvement of protein kinase C. In comparison, a high concentration of 1,25(OH)2 vitamin D3 (1,25D3) (1 x 10(-6) M) increased the L channel current in UMR 106 cells. Therefore, 24,25D3 appears to have an action on the L-channel current that is distinct from that of 1,25D3. This demonstration of a non-genomic effect of 24,25D3 on calcium channels suggests that 24,25D3 is an active metabolite of vitamin D3 and may play an important role in regulating the function of bone cells. PMID- 8732261 TI - Calcium imaging shows differential sensitivity to cooling and communication in luminous transgenic plants. AB - Imaging of a recombinant bioluminescent Ca2+ indicator, aequorin, in an entire organism showed three novel features of Ca2+ signals in plants. First, cooling the plant from 25 degrees C to 2 degrees C demonstrated differential sensitivities between organs, the roots firing a Ca2+ signal at some 8-10 degrees C higher than the cotyledons. Secondly, prolonged cooling provoked Ca2+ oscillations, but only in the cotyledons. These oscillations occurred with a frequency of 100 s and damped down within 800 s. Thirdly, cooling the roots of mature plants triggered a Ca2+ signal in the leaves, as a result of organ-organ communication. However, warming and then recooling the roots did not generate a second Ca2+ signal in these leaves. This desensitisation was not due to down regulation in the leaf since this was able to generate a Ca2+ signal of its own when cooled directly. Thus a combination of a recombinant bioluminescent indicator with photon counting imaging reveals startling new aspects of signalling in intact organs and whole organisms. PMID- 8732260 TI - Agonist-evoked calcium efflux from a functionally discrete compartment in Xenopus oocytes. AB - Agonist-induced calcium (Ca) mobilization is accompanied by Ca efflux, presumably reflecting the rise in Ca concentration at the cytosolic surface of the cell membrane. We studied the relationship between Ca efflux and intracellular Ca mobilization in Xenopus oocytes. Elevation of cytosolic Ca by a direct injection of 1 nmol 45CaCl2 resulted in a typical Ca-activated chloride current, but not in 45Ca efflux. This demonstrated that a Ca rise at the cytoplasmic surface of the membrane is not sufficient to produce an increased efflux. Co-injection of inositol 1,4,5-trisphosphate (InsP3), to prevent rapid Ca sequestration, also failed to cause Ca efflux. Smaller amounts of labelled Ca (0.05 nmol) equilibrated with Ca stores in a time-dependent pattern with an optimum at 2 h after injection. In contrast, Ca taken up from the medium was immediately available for agonist- or InsP3-induced efflux. Emptying the agonist-sensitive stores with thapsigargin (TG) did not affect chloride currents induced by Ca injection, indicating that these currents were due to direct elevation of Ca at the plasma membrane, rather than Ca-induced Ca release from InsP3-sensitive stores. Agonist-induced depletion of Ca stores enhanced uptake from the extracellular medium and the subsequent release of the label by an agonist. Similar protocol when the label was injected into the oocytes, failed to affect agonist induced efflux. We suggest that, under physiological conditions, agonist dependent Ca extrusion or uptake in oocytes is executed exclusively via a functionally restricted compartment, which is closely associated with both agonist-sensitive Ca stores and the plasma membrane. PMID- 8732262 TI - Spatial and temporal aspects of Ca2+ oscillations in Xenopus laevis melanotrope cells. AB - Spatio-temporal aspects of Ca2+ signaling in melanotrope cells of Xenopus laevis have been studied with confocal laser-scanning microscopy. In the whole-frame scanning mode, two major intracellular Ca2+ compartments, the cytoplasm and the nucleus, were visualized. The basal [Ca2+] in the nucleus appeared to be lower than that in the cytoplasm and Ca2+ oscillations seemed to arise synchronously in both compartments. The N-type channel blocker omega-conotoxin eliminated oscillations in both regions, indicating a strong coupling between the two compartments with respect to Ca2+ dynamics. Line-scanning mode, which gives higher time resolution, revealed that the rise phase of a Ca2+ oscillation is not a continuous process but consists of 3 or 4 discrete steps. Each step can be seen as a Ca(2+)-wave starting at the cell membrane and going through the cytoplasm at a speed of 33.3 +/- 4.3 microns/s. Before the Ca(2+)-wave enters the nucleus, a delay of 120.0 +/- 24.1 ms occurred. In the nucleus, the speed of a wave was 80.0 +/- 3.0 microns/s. Treatment with the Ca(2+)-ATPase inhibitor thapsigargin (1 MicroM) almost completely eliminated the apparent difference in the basal [Ca2+] in the cytoplasm and the nucleus, reduced the delay of a Ca(2+)-wave before entering the nucleus to 79.8 +/- 8.7 ms, and diminished the nuclear wave speed to 35.0 +/- 4.9 microns/s. These results indicate that a cytoplasmic thapsigargin sensitive ATPase near the nuclear envelope is involved in buffering Ca2+ before the Ca2+ wave enters the nucleus. After sensitizing IP3 receptors by thimerosal (10 microM) the speed of the cytoplasmic Ca(2+)-wave was increased to 70.3 +/- 3.6 microns/s, suggesting that IP3 receptors may be involved in the propagation of the cytoplasmic Ca2+ wave. Our results indicate that in melanotropes the generation and propagation of Ca2+ oscillations is a complex event involving influx of Ca2+ through N-type Ca2+ channels, propagation of the cytoplasmic Ca2+ wave through mobilization of intracellular stores and a regulated Ca2+ entry into the nucleus. We propose that Ca(2+)-binding proteins may act as a Ca2+ store for propagation of the wave in the nucleus. PMID- 8732263 TI - Characterisation of the voltage-activated calcium current in the marine ciliate Euplotes vannus. AB - We have isolated the early Ca current (ICa) from the whole cell current that activates upon depolarisations in the marine ciliate Euplotes vannus. The peak of ICa activated within 4.2 ms at depolarisations to 5 mV with an amplitude of 2.5 +/- 0.35 nA and was reduced to 1.0 +/- 0.14 nA (n = 5) when the extracellular Ca concentration was changed from 10 to 1 mM. The voltage-dependent activation curve was steeper and shifted to more negative values when external Ca2+ was replaced by Ba2+. The early inward current inactivated with a double-exponential time course including a fast and a slow component, and no inactivation was recorded with Ba2+. The time constants for the recovery from inactivation varied between 44 and 153 ms according to the depolarisation-dependent Ca influx. At the common resting potential of -25 mV, ICa was not steady-state inactivated; ICa half inactivated at -14.5 mV, and totally inactivated at -5 mV. ICa was inhibited by 10 mM extracellular Cd2+. The peptides omega-conotoxin-GVIA (20 microM), omega conotoxin-MVIIC (600 nM), omega-agatoxin-IVA (60 nM) and calciseptine (900 nM) did not block ICa. The benzothiazepine-derivative diltiazem (100 microM) and the dihydropiridine nifedipine (100 microM) inhibited 51% and 33% of ICa, respectively. The naphthalene sulfonamide W7 reduced ICa with an inhibition coefficient of 33 microM. PMID- 8732264 TI - Effect of cAMP elevating agents on carbachol-induced phosphoinositide hydrolysis and calcium mobilization in cultured canine tracheal smooth muscle cells. AB - The effects of increases in intracellular adenosine 3',5'-cyclic monophosphate (cAMP) on carbachol-induced generation of inositol phosphates (IPs) and increases in intracellular Ca2+ ([Ca2+]i) were investigated in canine cultured tracheal smooth muscle cells (TSMCs). The cAMP elevating agents, cholera toxin (CTX) and forskolin, induced concentration- and time-dependent cAMP formation with half maximal effects (-logEC50) at concentrations of 7.6 +/- 1.3 g/ml and 4.8 +/- 0.9 M, respectively. Forskolin caused a concentration-dependent inhibition of carbachol-induced increase in [Ca2+]i with half-maximal inhibition (-logEC50) at 5.2 +/- 0.7 M. Pretreatment of TSMCs with either CTX (10 micrograms/ml, 4 h), forskolin (10-100 microM, 30 min), or dibutyryl cAMP (1 mM, 30 min) inhibited carbachol-stimulated Ca2+ mobilization and IPs accumulation. The inhibitory effects of these agents produced both depression of the maximal response and a shift to the right of the concentration-response curve of carbachol without changing the EC50 values. After treatment with forskolin for 24 h, carbachol induced IPs accumulation and Ca2+ mobilization were close to those of control group. SQ-22536 [9-(tetrahydro-2-furanyl)-9H-purin-6-amine, 10 microM], an inhibitor of adenylate cyclase, and HA-1004 [N-(2-guanidinoethyl)-5 isoquinolinesulfonamide hydrochloride, 50 microM], an inhibitor of cAMP-dependent protein kinase (PKA), attenuated the ability of forskolin to inhibit carbachol induced IPs accumulation. Moreover, the inactive analogue of forskolin, 1,9 dideoxy forskolin, did not inhibit these responses evoked by carbachol, suggesting that activation of cAMP/PKA was involved in these inhibitory effects of forskolin. The KD and Bmax values of the muscarinic receptor (mAChR) for [3H] N-methyl scopolamine binding were not significantly changed by forskolin treatment for 30 min and 24 h, suggesting that the inhibitory effect of forskolin is distal to the mAChR. The locus of this inhibition was further investigated by examining the effect of forskolin treatment on AIF4(-)-stimulated IPs accumulation in canine TSMCs. The AIF4(-)-induced response was inhibited by forskolin, supporting the notion that G protein(s) are directly activated by AIF4 and uncoupled to phospholipase C by forskolin treatment. We conclude that cAMP elevating agents inhibit carbachol-stimulated generation of IPs and Ca2+ mobilization in canine cultured TSMCs. Since generation of IPs and increases in [Ca2+]i are very early events in the activation of mAChRs, attenuation of these events by cAMP elevating agents might well contribute to the inhibitory effect of cAMP on tracheal smooth muscle formation. PMID- 8732265 TI - Photolysis of caged compounds characterized by ratiometric confocal microscopy: a new approach to homogeneously control and measure the calcium concentration in cardiac myocytes. AB - Here we describe the subcellularly uniform control of the intracellular Ca2+ concentration ([Ca2+]i) by flash photolysis of caged Ca2+ or a caged Ca2+ buffer. A mixture of the two Ca2+ indicators Fluo-3 and Fura-red was used together with a laser-scanning confocal microscope to reveal spatial aspects of intracellular Ca2+ signals. The patch clamp technique in the whole-cell variant was applied to load the cells with the indicator mixture together with either DM-nitrophen or diazo-2 and to measure changes in the membrane current. An in vivo calibration was performed to convert the Fluo-3/Fura-red fluorescence ratios to [Ca2+] values. The resulting calibration curve suggested an apparent KD of 1.6 microM, Rmax of 2.15, Rmin of 0.08 and a Hill-coefficient of 0.75 for the indicator mixture. Controlled rupture of the cell membrane revealed a large fraction of immobile intracellular Fura-red fluorescence that may account for the reduced in vivo Rmax value when compared to the in vitro value of 3.1. In cardiac myocytes, flash photolytic release of Ca2+ from DM-nitrophen generated inwardly directed Na+/Ca2+ exchange currents and Ca2+ signals that were graded with the discharged flash-energy. Rapid line-scans revealed subcellularly homogeneous [Ca2+] jumps regardless of the discharged flash energy. Ca2+ signals evoked by L-type Ca2+ currents (ICa) could be terminated rapidly in a spatially homogeneous manner by UV flash photolysis of diazo-2. No side-effects of the photolytic products of DM nitrophen or diazo-2 with the mixture of Fluo-3/Fura-red were detectable in our experiments. The combination of UV flash photolysis and laser scanning confocal microscopy enabled us to control [Ca2+]i homogeneously on the subcellular level. This approach may improve our understanding of the subcellular properties of cardiac Ca2+ signalling. The technique can also be applied in other cell types and with other signalling systems for which caged compounds are available. PMID- 8732266 TI - Increase by the ORL1 receptor (opioid receptor-like1) ligand, nociceptin, of inwardly rectifying K conductance in dorsal raphe nucleus neurones. AB - The actions of the endogenous ORL1-receptor (opioid receptor-like1) ligand, nociceptin, on the membrane properties of rat dorsal raphe nucleus neurones were examined by use of whole-cell patch clamp recording in brain slices. Nociceptin produced an outward current in all neurones tested, with an EC50 of 12 +/- 2 nM. Dynorphin A (100 nM to 1 microM) produced little outward current. Outward currents reversed polarity near the predicted K+ equilibrium potential in both 2.5 mM (measured/predicted = -105 mV/-104 mV) and 6.5 mM (measured/predicted = 80 mV/-77 mV) extracellular K+. The conductance increase was larger between -120 and -130 mV than between -70 and -80 mV, conductance. The outward current produced by nociceptin was similar to, and occluded by, high concentrations of baclofen, demonstrating actions on the same population of K channels. Naloxone (1 microM) failed to inhibit outward currents produced by nociceptin. These results are consistent with the reported high density of ORL1 receptor mRNA in dorsal raphe nucleus and with inhibitory actions of nociceptin in cells expressing ORL1. PMID- 8732267 TI - Investigation into the 5-hydroxytryptamine-induced atropine-resistant neurogenic contraction of guinea-pig proximal colon. AB - 1. The aim of this study was to characterize the receptors mediating the atropine resistant neurogenic contraction to 5-hydroxytryptamine (5-HT) in the longitudinal muscle of the guinea-pig proximal colon and to determine the type of tachykinin receptors involved in the contractile response to 5-HT by the use of selective antagonists. 2. In the presence of atropine (0.3 microM), guanethidine (5 microM), hexamethonium (100 microM), ketanserin (0.1 microM) and indomethacin (3 microM), 5-HT (0.01-3 microM) produced concentration-dependent neurogenic contractions of colonic strips and at 0.3 microM produced a maximal effect (pEC50 = 7.39 +/- 0.09, n = 18). The 5-HT4 receptor stimulant, 5-methoxytryptamine (5 MeOT, 0.03-10 microM) also produced neurogenic contractions with similar maximum effect to those of 5-HT (pEC50 = 6.89 +/- 0.16). 3. The 5-HT4 receptor antagonist, DAU 6285 (3 microM) shifted the concentration-response curves to both 5-HT and 5-MeOT to the right without significant depression of the maximum, but the 5-HT1/5-HT2 receptor antagonist, metitepine (0.1 microM) and the 5-HT3 receptor antagonist, ondansetron (0.3 microM) had no effect on the control curves to 5-HT and 5-MeOT. 4. The selective NK1 receptor antagonist, FK 888 (1 microM) markedly attenuated the contractions to 5-HT and 5-MeOT. In contrast, the selective NK2 receptor antagonist, SR 48968 (10 nM) and the selective NK3 receptor antagonist, SR 142801 (10 nM) had no effect on the contractions to 5-HT and 5-MeOT. 5. These results indicate that the 5-HT-induced atropine-resistant neurogenic contraction of guinea-pig proximal colon is due to activation of 5-HT4 receptors, presumably located on excitatory motor neurones, innervating the longitudinal muscle. The contraction evoked by activation of the 5-HT4 receptors is mediated primarily via NK1 receptors but not NK2 or NK3, suggesting that the 5 HT4 receptor-mediated contraction is evoked indirectly via tachykinin release from tachykinin-releasing excitatory neurones. PMID- 8732268 TI - The longitudinal muscle of rat ileum as a sensitive monoreceptor assay for bradykinin B1 receptors. AB - 1. Various bradykinin derivatives, acting preferentially at B1 or B2 receptors, were tested in the isolated longitudinal smooth muscle of rat ileum. Experiments were carried out in the presence of chlorpheniramine and atropine (both 1 microM), guanethidine and indomethacin (both 3 microM) and of the peptidase inhibitors (captopril, bestatin and thiorphan, all 1 microM). 2. The rank order of potency was (pD2 values +/- s.e.mean, n = 5 in parentheses, at 5 h from set up): [des-Arg9]-BK (8.27 +/- 0.11) > or = [des-Arg10]-kallidin (7.67 +/- 0.24) > bradykinin (6.69 +/- 0.25). The B2 receptor selective agonist, [Hyp3,Tyr(Me)8] BK, was approximately 10 fold less active than bradykinin. Contractile responses to all agonists increased with time. The maximal response to the B1 receptor agonist, [desArg9]-BK at 5 h (94 +/- 2%) was significantly (P < 0.05) greater than that measured at 2 h (74 +/- 2%). 3. The B2 receptor antagonist, D-Arg[Hyp3, Thi5, D-Tic7, Oic8]-BK (Hoe 140, 0.1 microM) did not affect responses to the B1 receptor agonist [des-Arg9]-BK (0.1 nM--1 microM) nor those to the B2 receptor agonist, [Hyp3,Tyr(Me)8]-BK (1 nM--10 microM). In control experiments performed in the longitudinal smooth muscle of guinea-pig ileum and rat isolated urinary bladder as bioassays for B2 receptors, the B2 receptor antagonist Hoe 140 (0.1 microM) antagonized bradykinin-induced contractions. 4. In the rat isolated ileum the B1 receptor antagonist, D-Arg[Hyp3, Thi5, D-Tic7, Oic8, des-Arg9]-BK ([des Arg10]-Hoe 140, 0.3 - 10 microM) competitively antagonized contractile responses to [des-Arg9]-BK with an estimated pKB of 6.74 +/- 0.08 (Schild plot slope with confidence limits 1.22, (0.70 - 1.73) n = 13). In control experiments in the guinea-pig isolated ileum and rat isolated urinary bladder, [des-Arg10]-Hoe 140 (1 - 10 microM) did not inhibit B2 receptor-mediated contractile responses. 5. The putative B1 receptor antagonist, [Leu8,des-Arg9]-BK, behaved as a partial agonist when responses were determined 2 h from set-up (pD2 6.43 +/- 0.21, n = 5; Emax 30% of that evoked by [des-Arg9]-BK); at 5 h from set-up it behaved as a full agonist (pD2 7.48 +/- 0.12, n = 5; Emax 90% of that evoked by [des-Arg9] BK). At this time the response to [Leu8,des-Arg9]-BK was antagonized in a concentration-dependent manner by [des-Arg10]-Hoe 140, which at 1 microM and 10 microM, produced dose-ratios of 6.33 +/- 3.66 (n = 4) and 103 +/- 40 (n = 4). 6. In view of the rank order of potency of agonists, the antagonist activity by [des Arg10]-Hoe 140 and the lack of antagonist activity of Hoe 140, we conclude that the longitudinal smooth muscle of rat ileum, after histamine, acetylcholine, noradrenaline, and prostanoid production blockade, is a sensitive monoreceptor assay for studying the pharmacology of bradykinin B1 receptors. Further the preparation can also be used as a sensitive bioassay to identify partial agonist activity of B1 receptor antagonists such as [Leu8,desArg9]-BK. PMID- 8732269 TI - In vitro pharmacological profile of YM-43611, a novel D2-like receptor antagonist with high affinity and selectivity for dopamine D3 and D4 receptors. AB - 1. We investigated some neurochemical properties of a novel benzamide, YM-43611, [(S)-N-(1-benzyl-3-pyrrolidinyl)-5-chloro-4-cyclopropylcarbonylamino+ ++-2- methoxybenzamide] in comparison with putative D2-like receptor antagonists using both rat and human cloned dopamine D2-like receptors in vitro. 2. Receptor binding studies revealed that YM-43611 had appropriately potent affinities for both rat and human D2-like receptors, with moderate selectivity for D3 receptors and high selectivity for D4 receptors over D2 receptors (Ki values (nM) for rat receptors: D2, 165; D3, 35.5; D4, 1.85, and for human receptors: D2, 42.9; D3, 11.2; D4, 2.10). 3. YM-43611 displayed weak or negligible affinity for other neurotransmitter receptors, namely D1, D5, alpha(1), alpha(2), beta, 5-HT1A, 5 HT2A, 5-HT3, H1, M1 and M2 receptors. 4. Dopamine stimulated low-Km GTPase activity on membranes from Chinese hamster ovary (CHO) cells expressing the human D2-like receptor subtype. This response to dopamine of low-Km GTPase activity was inhibited by use of putative D2-like receptor antagonists. YM-43611 showed a moderate selectivity for D3 receptors (Ki = 45.5 nM) and a high selectivity for D4 receptors (Ki = 3.28 nM) over D2 receptors (Ki = 70.6 nM). 5. Dopamine inhibited forskolin-stimulated adenylate cyclase in intact CHO cells expressing the human D2-like receptor subtype. YM-43611 shifted the inhibition curve of dopamine on respective D2-like receptor subtype-mediated cyclic AMP formation to the right in a parallel fashion, showing a pA2 value of 7.42 (38.1 nM) for D2 receptors, a pKB value of 8.06 (8.68 nM) for D3 receptors, and a pA2 value of 8.42 (3.77 nM) for D4 receptors. 6. YM-43611 but not the other D2-like receptor antagonists exhibited good selectivity with respect to dual antagonism for D3 and D4 receptors in both receptor binding and functional assays. 7. These results indicate that YM-43611 is a novel D2-like receptor antagonist with high potency and selectivity for both D3 and D4 receptors. YM-43611 is therefore expected to be valuable in exploration of the physiological role of D3 and D4 receptors. PMID- 8732270 TI - Blockade of nitrergic transmission by hydroquinone, hydroxocobalamin and carboxy PTIO in bovine retractor penis: role of superoxide anion. AB - 1. The effects of inhibiting endogenous Cu/Zn superoxide dismutase (SOD) with diethyldithiocarbamate (DETCA) were examined on the ability of hydroquinone, hydroxocobalamin and carboxy-PTIO to block nitrergic relaxation in the bovine retractor penis (BRP) muscle. 2. Incubation of strips of BRP with DETCA (3 mM) for 2 h reduced SOD activity from 73.1 +/- 15.7 to 8.2 +/- 1.9 units mg-1 protein. 3. Hydroquinone (10 microM--1 mM) produced weak inhibition of nitrergic (4 Hz, 10 s) relaxation in control strips of BRP, but powerful inhibition in strips treated with DETCA (3 mM, 2 h). Exogenous SOD (250 units ml--1) produced a partial blockade of the ability of hydroquinone to inhibit nitrergic relaxation in DETCA-treated strips. 4. In an assay of SOD-inhibitable reduction of cytochrome C, hypoxanthine (0.1 mM)/xanthine oxidase (16 munits ml-1) and pyrogallol (10 microM), led to the rapid generation of superoxide anion. Hydroquinone (10 microM) also led to the generation of the free radical, although the rate of generation was slower. 5. Two NO-scavenging agents, hydroxocobalamin (0.1 microM--1 mM) and carboxy-PTIO (0.1-1 mM), produced concentration-dependent blockade of nitrergic relaxation of the BRP. The magnitude of the blockade induced by these agents was unaffected following treatment with DETCA or SOD. 6. The findings with hydroquinone support our previous proposal that endogenous Cu/Zn SOD plays a vital role in protecting nitrergic neurotransmission from inactivation by superoxide anion. Results with hydroxocobalamin and carboxy-PTIO are consistent with the known ability of these agents to scavenge NO. The nitrergic neurotransmitter in the BRP thus appears to have the properties of NO. PMID- 8732271 TI - Adenosine A2 receptor-induced inhibition of leukotriene B4 synthesis in whole blood ex vivo. AB - 1. Engagement of adenosine A2 receptors suppresses several leukocyte functions. In the present study, we examined the effect of adenosine on the inhibition of leukotriene B4 (LTB4) synthesis in heparinized human whole blood, pretreated with lipopolysaccharide (LPS) and tumour necrosis factor alpha (TNF-alpha) and stimulated with the chemotactic peptide, N-formyl-Met-Leu-Phe (FMLP). 2. The FMLP induced synthesis of LTB4 in whole blood pretreated with LPS and TNF-alpha was dose-dependently inhibited by adenosine analogues in the following order of potency; 5'(N-ethyl)carboxamidoadenosine (NECA) approximately equal to CGS 21680 > 2-Cl-adenosine > N6-cyclopentyladenosine (CPA), indicating the involvement of the adenosine A2 receptor subtype. The IC50 values for NECA, CGS 21680, 2-Cl adenosine, and CPA were 6 nM, 9 nM, 180 nM, and 990 nM, respectively. 3. Dipyridamole, an agent that blocks the cellular uptake of adenosine by red cells and causes its accumulation in plasma, also inhibited the synthesis of LTB4 in LPS and TNF-alpha-treated whole blood stimulated by FMLP; moreover, this inhibition was reversed upon addition of adenosine deaminase. 4. A highly selective antagonist of the adenosine A2 receptor, 8-(3-chlorostyryl)caffeine (CSC), reversed the inhibition of LTB4 synthesis by 2-Cl-adenosine and dipyridamole in LPS and TNF-alpha-treated whole blood, stimulated by FMLP. 5. LTB4 synthesis in whole blood originates predominantly from neutrophils and to a lesser extent from monocytes. 2-Cl-adenosine also inhibited the synthesis of LTB4 induced by FMLP in these isolated LPS and TNF-alpha-treated cells; however, 2-Cl adenosine was a more potent inhibitor of LTB4 synthesis in neutrophils than monocytes. 6. The present data demonstrate that adenosine, acting through A2 receptors, exerts a potent inhibitory effect on the synthesis of LTB4 and thus contribute to the understanding of its anti-inflammatory properties. PMID- 8732272 TI - KW-3902, a selective high affinity antagonist for adenosine A1 receptors. AB - 1. We demonstrate that 8-(noradamantan-3-yl)-1,3-dipropylxanthine (KW-3902) is a very potent and selective adenosine A1 receptor antagonist, assessed by radioligand binding and cyclic AMP response in cells. 2. In rat forebrain adenosine A1 receptors labelled with [3H]-cyclohexyladenosine (CHA), KW-3902 had a Ki value of 0.19 nM, whereas it showed a Ki value of 170 nM in rat striatal A2A receptors labelled with [3H]-2-[p-(2-carboxyethyl)-phenethylamino]-5'-N ethylcarboxamidoad enosine (CGS21680), indicating 890 fold A1 receptor selectivity versus the A2A receptor. KW-3902 at 10 microM showed no effect on recombinant rat A3 receptors expressed on CHO cells. 3. Saturation studies with [3H]-KW-3902 revealed that it bound with high affinity (Kd = 77 pM) and limited capacity (Bmax = 470 fmol mg-1 of protein) to a single class of recognition sites. A high positive correlation was observed between the pharmacological profile of adenosine ligands inhibiting the binding of [3H]-KW-3902 and that of [3H]-CHA. 4. KW-3902 showed potent A1 antagonism against the inhibition of forskolin-induced cyclic AMP accumulation in DDT1 MF-2 cells by the A1-selective agonist, cyclopentyladenosine with a dissociation constant (KB value) of 0.34 nM. KW-3902 antagonized 5'-N-ethylcarboxamidoadenosine-elicited cyclic AMP accumulation via A2B receptors with a KB value of 52 nM. 5. KW-3902 exhibited marked species-dependent differences in the binding affinities. The highest affinity was for the rat A1 receptor (ki = 0.19 nM) and these values for guinea pig and dog A1 receptors were 1.3 and 10 nM, respectively. PMID- 8732273 TI - Effects of some K(+)-channel inhibitors on the electrical behaviour of guinea-pig isolated trachealis and on its responses to spasmogenic drugs. AB - 1. A study has been made of the effects of inhibitors selective among plasmalemmal K(+)-channels on the sensitivity and responsiveness of guinea-pig trachealis muscle to carbachol, histamine and KCl. The effects of the K(+) channel inhibitors on the resting membrane potential and spontaneous electrical activity of the trachealis cells have also been examined. 2. In indomethacin (2.8 microM)-treated trachealis muscle, dofetilide (1 microM) and glibenclamide (10 microM) were each devoid of spasmogenic activity. In contrast, 4-aminopyridine (4 AP, 62.5 microM--8 mM), charybdotoxin (ChTX, 100 nM) and iberiotoxin (IbTX, 100 nM) were each spasmogenic. Spasm evoked by 4-AP, IbTX or ChTX was reduced, though not abolished, by atropine (1 microM). Spasm evoked by 4-AP (1 mM), ChTX (100 nM) or IbTX (100 nM) was unaffected by tetrodotoxin (TTX; 3.1 microM) or by tissue pretreatment with capsaicin (1 microM for 30 min). Spasm evoked by IbTX or ChTX was abolished by nifedipine (1 microM). 3. Dofetilide (1 microM) and glibenclamide (10 microM) were each without effect on the tracheal sensitivity or responsiveness to carbachol, histamine or KCl. 4-AP (1 mM) antagonized carbachol, potentiated histamine but did not affect tissue sensitivity to KCl. When the effects of 4-AP were examined in the presence of atropine (1 microM), it potentiated all the spasmogens including carbachol. IbTX and ChTX (each 100 nM) potentiated all three spasmogens. Potentiation of histamine induced by 4-AP (1 mM) or IbTX (100 nM) was also observed in tissues treated with a combination of atropine (1 microM) and TTX (3.1 microM). 4. Dofetilide (1 and 10 microM) was without effect on the resting membrane potential or spontaneous electrical activity of the trachealis cells. 4-AP (1 mM) evoked depolarization and caused a small increase in the frequency of slow wave discharge. The depolarization evoked by 4-AP was abolished by atropine (1 microM). IbTX (100 nM) and ChTX (100 nM) each evoked little or no change in resting membrane potential but converted the spontaneous slow waves into spike-like, regenerative action potentials. These electrophysiological effects of IbTX and ChTX were unaffected by atropine (1 microM). 5. It is concluded that the dofetilide-sensitive, cardiac, delayed rectifier K(+)-channel is either not expressed in trachealis muscle or is of no functional importance in that tissue. The ATP-sensitive K(+)-channel (KATP) does not moderate tracheal sensitivity to spasmogens such as carbachol, histamine and KCl. The 4-AP-sensitive delayed rectifier K(+)-channel (Kdr) and the large Ca(2+) dependent K(+)-channel (BKCa) each moderate trachealis muscle sensitivity to spasmogens. Neither Kdr nor BKCa plays an important role in determining the resting membrane potential of guinea-pig trachealis cells. However, the BKCa channel is responsible for limiting the effects of the increase in membrane Ca2+ conductance associated with the depolarizing phase of slow waves. It is BKCa channel opening that prevents the development of a slow wave into a spike-like regenerative action potential. PMID- 8732274 TI - Allosteric interactions between cyclothiazide and AMPA/kainate receptor antagonists. AB - 1. Cyclothiazide blocks alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor desensitization and potentiates AMPA receptor gated currents. Interactions between cyclothiazide, and the non-competitive antagonist GYKI52466 (GYKI) and competitive antagonist 2,3-dihydroxy-6-nitro-7-sulphamoyl-benzo (F) quinoxaline (NBQX) were studied at native and recombinant AMPA/kainate receptors using whole-cell recording in order to characterize the modulation by cyclothiazide of these two antagonist sites. 2. GYKI 100 microM, which is sufficient to eliminate virtually hippocampal kainate (100 microM) currents, failed to prevent access of cyclothiazide to its site of potentiation, and was unable to enhance removal of cyclothiazide potentiation. However, cyclothiazide reduced GYKI (30 microM) block from 84 +/- 8.3% to 38 +/- 12%, and slowed the onset of the block with a time course much faster than the time course for onset and offset of potentiation induced by cyclothiazide. Cyclothiazide had qualitatively similar effects upon antagonism by NBQX 1 microM. 3. Kainate activated desensitizing currents in dorsal root ganglion (DRG) neurones, which were unaffected by cyclothiazide. GYKI blocked these kainate currents with lower affinity (IC50 > 120 microM) than for hippocampal neurones (IC50 < 30 microM), and cyclothiazide did not affect GYKI antagonism. 4. Steady-state AMPA currents from homomeric GluRA-Dflip receptors in HEK 293 cells were dramatically potentiated (up to 216 fold) by cyclothiazide via reduction of desensitization. In contrast, kainate-gated currents in HEK 293 cells expressing GluR6R receptors exhibited pronounced desensitization that was unaffected by cyclothiazide. GYKI retains its inhibition at both recombinant AMPA and kainate receptors. 5. These results indicate that cyclothiazide allosterically influences two important antagonist sites on AMPA receptors. In addition, AMPA/kainate receptor subunit composition influences the affinity of GYKI for the receptor. PMID- 8732275 TI - Operational characteristics of somatostatin receptors mediating inhibitory actions on rat locus coeruleus neurones. AB - 1. In order to characterize somatostatin (SRIF) receptor inhibiting spontaneous firing of rat locus coeruleus neurones, and their transduction mechanism(S), extracellular recordings were obtained from a pontine slice preparation of rat brain containing the locus coeruleus (LC). LC neurones were identified by electrophysiological and pharmacological properties; spontaneous firing (characteristically 0.5-5 Hz) was reversibly and concentration-dependently inhibited by exogenously applied noradrenaline. 2. Spontaneous firing of LC neurones was reversibly and concentration-dependently inhibited by SRIF and the N terminally extended form, somatostatin-28 (SRIF-28), with EC50 values of 15.1 and 19.4 nM, respectively. The synthetic SRIF analogues (octreotide, MK-678, BIM 23027 and L-362,855) also caused concentration-dependent inhibition of LC neurone firing with a rank order of agonist potencies compatible with actions at a receptor resembling the recombinant sst2 receptor. The putative sst3 selective agonist, BIM-23056, was without agonist or antagonist effect. 3. Addition of 100 nM desipramine significantly increased the efficacy of exogenously applied noradrenaline (EC50 values, 2.96 and 0.13 microM, absence and presence of desipramine, respectively) but did not significantly affect SRIF-induced inhibition (EC50 values, 15.6 and 8.0 nM, respectively). Furthermore, application of phenoxybenzamine (3 microM) abolished responses to NA, but did not affect responses to SRIF (EC50 = 14.1 nM). 4. Application of the cyclic AMP analogue, 8 bromoadenosine-cyclic monophosphate (8-Br-cyclic AMP; 500 microM), significantly increased the spontaneous firing rate of all neurones tested (223 +/- 24% over basal rate). Concentration-effect curves for SRIF constructed in the absence and presence of 8-Br-cyclic AMP had similar threshold concentrations, maxima and EC50 values. 5. Incubation of pontine slices in a modified artificial CSF containing 500 ng ml-1 pertussis toxin (PTX) for 18 h prior to extracellular recording affected neither the spontaneous firing of LC neurones, nor the inhibitory responses to muscimol (EC50 2.2 and 1.2 microM, absence and presence of PTX). However, inhibitory responses to SRIF were markedly attenuated. 6. We conclude that the inhibitory actions of SRIF on spontaneous firing of LC neurones are mediated directly by activation of somatodendritic SRIF receptors, and not indirectly by release of noradrenaline. The SRIF receptors involved appear to couple via a pertussis toxin sensitive G-protein, and elicit their response by a mechanism apparently independent of inhibition of cyclic AMP formation. The agonist profile of several selective and novel SRIF analogues suggests the identity of this receptor to be similar to the recombinant sst2 receptor. PMID- 8732276 TI - 5-HT3 and 5-HT4 receptor-mediated facilitation of the emptying phase of the peristaltic reflex in the marmoset isolated ileum. AB - 1. The patterns of intestinal motility and the effect of an increase in intraluminal pressure were studied in vitro on segments obtained from the marmoset small intestine. 2. Segments obtained from the distal half of the marmoset small intestine exhibited segmentation, consisting of narrow annular contractions (lasting for 2-3 s) interposed between the relaxed segments of varying length. The subsequent contractions occurred slightly distal to the previous contraction, with ring-like contractions appearing to move in the aboral direction. Such segmentation was infrequent or absent in the segments obtained from the proximal half of the small intestine. An increase in intraluminal pressure inhibited segmentation and finally produced peristalsis in most of the tissues. 3. The influence of 5-hydroxytryptamine (5-HT) receptor agonists and antagonists on the threshold of the peristaltic reflex was investigated in the segments obtained from the distal half of the intestine after segmentation had subsided. The effect of drug application to the serosal surface was measured as a change in threshold pressure required to trigger the peristaltic reflex. A facilitation was defined in two ways (a) as a reduction in the threshold pressure required to trigger the reflex and (b) in those tissues that failed to respond with peristalsis on raising intraluminal pressure to the maximum attainable (1 kPa), as a reduction in threshold pressure compared to this value. 4. 5-HT (7.85 +/- 0.19), 5-methoxytryptamine (7.79 +/- 0.24), 5-carboxamidotryptamine (6.66 +/- 0.13) and 2-methyl-5-HT (6.24 +/- 0.16) caused a concentration related facilitation of the peristaltic reflex, the pD2 values (mean +/- s.e.mean) being shown in parentheses. 5. The concentration-response curves to both 5-HT and 5 methoxytryptamine were dextrally shifted in a surmountable manner in the presence of GR 113808 (30 nM). pD2 values for 5-HT and 5-methoxytryptamine were significantly decreased to 6.98 +/- 0.24 and 6.83 +/- 0.36 respectively in the presence of GR 113808 (30 nM). 6. In the presence of a high concentration of (10 microM) 5-methoxytryptamine the subsequent addition of 2-methyl-5-HT (3-10 microM) but not 5-methoxytryptamine (10 microM) facilitated peristalsis; the effect of 3 microM 2-methyl-5-HT was significantly decreased by 2 microM ondansetron. 7. It is concluded that the facilitation of the peristaltic reflex in the marmoset intestine induced by 5-HT at submicromolar concentrations involves a 5-HT4 receptor stimulation with an additional 5-HT3 receptor activation at higher concentrations. PMID- 8732277 TI - Myocardial protection after monophosphoryl lipid A: studies of delayed anti ischaemic properties in rabbit heart. AB - 1. Monophosphoryl lipid A (MLA) is a non-pyrogenic derivative of Salmonella lipopolysaccharide. Administration of this agent at high doses to rats and at low doses to dogs was previously shown to confer marked protection against ischaemia reperfusion 24 h later, although the cellular mechanisms of this delayed protection are obscure. We hypothesized that MLA pretreatment causes the induction of the 70 kDa cytoprotective stress protein HSP70i in the myocardium. If this were the case, protection against ischaemia-reperfusion injury would be observed both in vitro and in vivo. 2. Rabbits were pretreated with MLA 0.035 mg kg-1, i.v. or vehicle solution. For the in vitro study, hearts were isolated 24 h later and Langendorff-perfused with Krebs-Henseleit buffer at 37 degrees C. Global ischaemia was induced for 20 min followed by 120 min reperfusion. Recovery of post-ischaemic left ventricular function and lactate dehydrogenase efflux was similar in MLA and vehicle pretreated hearts and there was no significant difference in the percentage of infarction of the left ventricle determined by triphenyltetrazolium staining (MLA 22.4 +/- 5.2%, vehicle 24.8 +/- 5.1%). 3. When 30 min regional ischaemia and 120 min reperfusion was instituted in pentobarbitone-anaesthetized rabbits 24 h after pretreatment with MLA or vehicle, the percentage infarction within the risk zone was reduced from 42.6 +/- 5.7% in vehicle pretreated animals to 19.6 +/- 4.4% in MLA pretreated animals (P < 0.01). 4. Determination of myocardial HSP70i content by Western blot analysis showed that MLA treatment did not increase HSP70i immunoreactivity. 5. We conclude that MLA at this dose confers protection only against ischaemia-reperfusion injury in vivo and that this protection is not related to induction of HSP70i. Because protection was observed only in vivo it seems possible that the delayed protection conferred by MLA is mediated by effects on humoral or blood-borne factors. PMID- 8732278 TI - Pharmacological and biochemical characterization of purified A2a adenosine receptors in human platelet membranes by [3H]-CGS 21680 binding. AB - 1. The binding properties of human platelet A2a adenosine receptors, assayed with the A2a-selective agonist, [3H]-2-[p-(2-carboxyethyl)-phenethylamino]-5'-N ethylcarboxamidoad enosine ([3H]-CGS 21680), are masked by a non-receptorial component, the adenotin site. In order to separate A2a receptors from adenotin sites, human platelet membranes were solubilized with 1% 3-[(3 cholamidopropyl)dimethyl-ammonio]-1-propanesulphonate (CHAPS). The soluble platelet extract was precipitated with polyethylene glycol (PEG) and the fraction enriched in adenosine receptors was isolated from the precipitate by differential centrifugation. 2. The present paper describes the binding characteristics of the selective A2a agonist, [3H]-CGS 21680, to this purified platelet membrane preparation. In addition, receptor affinity and potency of several adenosine agonists and antagonists were determined in binding and adenylyl cyclase studies. 3. Saturation experiments revealed a single class of binding site with Kd and Bmax values of 285 nM and 2.07 pmol mg-1 of protein respectively. Adenosine receptor ligands competed for the binding of 50 nM [3H]-CGS 21680 to purified protein, showing a rank order of potency consistent with that typically found for interactions with the A2a adenosine receptors. In the adenylyl cyclase assay the compounds examined exhibited a rank order of potency very close to that observed in binding experiments. 4. Thermodynamic data indicated that [3H]-CGS 21680 binding to the purified receptor is totally entropy-driven in agreement with results obtained in rat striatal A2a adenosine receptors. 5. It is concluded that in the purified platelet membranes there is a CGS 21680 binding site showing the characteristic properties of the A2a receptor. This makes it possible to use this compound for reliable radioligand binding studies on the A2a adenosine receptor of human platelets. PMID- 8732279 TI - Effect of a non-sulphonylurea hypoglycaemic agent, KAD-1229 on hormone secretion in the isolated perfused pancreas of the rat. AB - 1. We examined the cooperative effect of a newly synthesized oral hypoglycaemic agent, KAD-1229 with glucose on insulin, glucagon and somatostatin secretion in the isolated perfused pancreas of the rat. 2. KAD-1229 stimulated concentration dependently the first phase of insulin secretion without the second phase in the presence of 2.8 mM glucose, while it stimulated both the first and the second phase of insulin release in the presence of 5.6 mM glucose. It was confirmed that the first phase of insulin release is depolarization-induced release with no other additional signal transduction. 3. KAD-1229 also enhanced insulin release evoked by 16.7 mM glucose, a concentration known to inhibit the ATP-sensitive K+ current completely. 4. A low concentration (2.8 mM) of glucose stimulated somatostatin release transiently, while a higher concentration (16.7 mM) of glucose exerted a sustained stimulation. KAD-1229 stimulated somatostatin secretion in a concentration-dependent manner irrespective of glucose concentrations. 5. When glucagon release was stimulated with 2.8 mM glucose, KAD 1229 inhibited this hypoglycaemia-induced glucagon secretion. 6. When pancreata from rats pretreated with streptozotocin (STZ) 60 mg kg-1 were perfused, the basal secretion of glucagon was markedly elevated, and the glucagon response to the low glucose was abolished. Further, the insulin and somatostatin responses to KAD-1229 were largely attenuated. KAD-1229 showed transient enhancement followed by inhibition of the glucagon release from the STZ-pretreated rat pancreas. 7. We conclude that KAD-1229 stimulates insulin and somatostatin release, while it inhibits glucagon release following transient stimulation. PMID- 8732280 TI - Some effects of nipradilol, a beta-antagonist possessing a nitroxy group, on smooth muscle of the pig coronary artery. AB - 1. The effects of nipradilol, a beta-adrenoceptor antagonist which possesses a nitroxy group, on cytosolic Ca2+ concentration ([Ca2+]i), and on tension development were simultaneously measured by front-surface fluorometry and fura-2 loaded strips in the proximal portion of pig coronary arteries. 2. Nipradilol reduced in a concentration-dependent manner both the [Ca2+]i and tension, irrespective of whether the strips were unstimulated or exposed to either high K+ or histamine containing solutions. However, both in the case of contractions induced by high K+-depolarization and histamine stimulation, for a given [Ca2+]i elevation the tension which developed in the presence of nipradilol was smaller than that generated in its absence, so that the [Ca2+]i-tension curves during the contraction were shifted to the right. 3. In the absence of extracellular Ca2+, the [Ca2+]i elevation due to the release of Ca2+ from histamine-sensitive store was inhibited by nipradilol. Nipradilol had no effect on the [Ca2+]i elevation due to the release of Ca2+ from caffeine-sensitive stores; however, it did inhibit the caffeine-induced increase in tension. A derivative of nipradilol, which lacked a nitroxy molecule (Nip(-N)), had no effect on the [Ca2+]i and tension elevated by histamine or caffeine in the absence of extracellular Ca2+. 4. The beta-adrenoceptor agonist, isoprenaline, reduced [Ca2+]i tension when applied to steady state contractions induced by high K+, or at the peak level of tension to histamine. The reduction of [Ca2+]i and tension induced by isoprenaline was inhibited by Nip(-N) in a concentration-dependent manner and nipradilol inhibited the isoprenaline-induced relaxation with bell-shaped concentration-response curves. At lower concentrations, nipradilol acted as a beta-blocker, the IC50- value being smaller than that of Nip(-N), and at higher concentrations, it acted as a nitrovasodilator. 5. Thus, it is suggested that, at lower concentrations, nipradilol, an antianginal drug, acts as a beta adrenoceptor antagonist. At higher concentrations, it relaxes the proximal portion of the coronary artery by directly reducing [Ca2+]i and the Ca2+ sensitivity of the myofilaments, apparently due to the presence of the nitroxy molecule. PMID- 8732281 TI - Ouabain-induced increases in resting tone of human hyperplastic prostate following repeated noradrenaline and electrical field stimulation. AB - 1. The effect of ouabain on contractions to repeated noradrenaline stimulation and electrical field stimulation of human hyperplastic prostate was examined. Ouabain (1 microM) did not induce contractile response per se but progressively increased the resting tone (i.e., the tone between one noradrenaline stimulation, or electrical field stimulation, and the following) of human hyperplastic prostate. 2. The increased tone by ouabain following repeated noradrenaline stimulations or electrical field stimulation was fully relaxed by the removal of external calcium, and recovered following restoration of calcium. 3. The effect of noradrenaline on NA+ uptake was measured. Noradrenaline (10 microM) significantly increased the rate of Na+ accumulation in the presence of ouabain (1 microM); this stimulatory effect was almost completely blocked by prazosin (0.1 microM) and ethylisopropylamiloride (100 microM). In contrast, tetrodotoxin (1 microM) had no effect on noradrenaline-stimulated Na+ transport in human hyperplastic prostate. 4. Intracellular Na+ loading by noradrenaline (10 microM) in the presence of ouabain (1 microM) significantly increased the transmembrane Ca2+ uptake as compared with the absence of ouabain; however, nifedipine (1 microM) was ineffective on Ca2+ uptake under this condition. 5. Transmembrane CA2+ efflux was stimulated by noradrenaline (10 microM) in human hyperplastic prostate; this effect was significantly decreased in the presence of ouabain (1 microM). 6. It is suggested that the increased tone of human hyperplastic prostate following repeated excitation in the presence of ouabain is due to increased Ca2+ entry and reduced efflux of Ca2+ through the Na+/Ca+ exchange system as a consequence of Na+ pump inhibition by ouabain. PMID- 8732282 TI - P2-purinoceptors mediating spasm of the isolated uterus of the non-pregnant guinea-pig. AB - 1. The isolated uterus of the non-pregnant guinea-pig has been suggested to contain P1-, and possibly P2-purinoceptors mediating spasm. The presence of P1 purinoceptors has been confirmed and these receptors have been further characterized. 2. In the presence of the adenosine uptake inhibitor, S-(4 nitrobenzyl)-6-thioinosine (NBTI, 300 nM) and a pA100 concentration of the P1 purinoceptor antagonist 8-sulphophenyltheophylline (140 microM), the potency order of agonists as spasmogens was: 2 methylthio ATP >> alpha,beta methylene ATP = UTP = ATP >> beta,gamma methylene ATP. This order is not consistent with any single recognised P2-purinoceptor subtype. 3. Indomethacin (1 microM) treatment abolished responses to 2 methylthio ATP, alpha,beta methylene ATP and UTP, while spasm to ATP was significantly inhibited. When the endometrial and circular smooth muscle cell layers were removed, spasmogenic responses to ATP, 2 methylthio ATP, alpha,beta methylene ATP and UTP were significantly reduced. 4. 2 methylthio ATP was able to cause desensitization to itself, but not to UTP, indicating that these agonists act at different receptor sites. 5. The P2 purinoceptor antagonist, suramin antagonized 2 methylthio ATP with a PA2 of 5.9 +/- 0.3. Suramin was also an antagonist of ATP and UTP. In the case of ATP, the antagonism was not dependent on suramin concentration, while for UTP the interaction appeared to be non-equilibrium. Pyridoxalphosphate-6-azophenyl-2',4' disulphonic acid (PPADS, 10 microM) had no effect on spasm to ATP, UTP or 2 methythio ATP. 6. In the presence of indomethacin, responses to ATP were unaffected by 8-sulphophenyltheophylline (140 microM) or by suramin (100 microM), but PPADS (10 microM) antagonized ATP. 7. These results suggest that the isolated uterus of the non-pregnant guinea-pig contains a mixture of P2-purinoceptors. P2U (or UTP-selective pyrimidinoceptors) and P2Y-purinoceptors appear to be present, probably mainly located on the endometrial or circular smooth muscle layer. Activation of these receptors leads to spasm via increases in prostanoid generation. There appears also to be a third class of non-P2X-, non p2Y purinoceptor present, at which ATP is an agonist and PPADS is an antagonist, located on the longitudinal smooth muscle, activation of which causes spasm independent of changes in prostanoids. PMID- 8732283 TI - Metabolism of the macrolide immunosuppressant, tacrolimus, by the pig gut mucosa in the Ussing chamber. AB - 1. The macrolide tacrolimus (FK506), used as an immunosuppressant, is a cytochrome P450 (CYP) 3A substrate in the liver. The metabolism of tacrolimus and the transport of its metabolites in the pig gut was studied in the Ussing chamber. Tacrolimus and its metabolites were quantified by h.p.l.c./mass spectrometry. 2. In the Ussing chamber, demethyl, didemethyl, hydroxy and hydroxy demethyl tacrolimus were generated. Their formation was concentration- and time dependent. The metabolite pattern was not different from that after incubation of tacrolimus with human small intestinal microsomes. 3. The metabolite formation was highest in the duodenum and declined in the order duodenum > jejunum > ileum > colon > stomach. 4. Since tacrolimus metabolism was inhibited by the specific CYP3A inhibitors, troleandomycin and ketoconazole, we concluded that these enzymes are involved in intestinal metabolism of tacrolimus. 5. Tacrolimus metabolites re-entered the mucosa chamber (> 90%) and passed through the small intestinal preparation into the serosa chamber. 6. It is concluded that tacrolimus is metabolized in the intestine, that the metabolites are able to re enter the gut lumen and also enter into the portal vein and that small intestinal metabolism and transport is at least in part responsible for the low oral bioavailability of tacrolimus. PMID- 8732284 TI - Differences in agonist and antagonist activities for two indices of metabotropic glutamate receptor-stimulated phosphoinositide turnover. AB - 1. The abilities of the four diastereoisomers of 1-aminocyclopentane-1,3 dicarboxylic acid (ACPD) to stimulate, and the metabotropic glutamate receptor (mGluR) antagonist (+/-)-alpha-methylcarboxyphenylglycine (MCPG) to inhibit, phosphoinositide turnover in neonatal rat cerebral cortex have been studied. Two indices of phosphoinositide cycle activity were assessed; inositol 1,4,5 trisphosphate (Ins(1,4,5)P3) mass accumulation, and total inositol phosphate [3H] InsPx accumulation (in the presence of Li+) in myo-[3H]-inositol prelabelled slices. 2. The diastereoisomers of ACPD stimulated each response with a rank order of potency of 1S, 3R > 1R, 3R > 1S, 3S >> 1R, 3S. The response to 1R, 3R ACPD was largely prevented by pre-addition of the NMDA-receptor antagonist, MK 801, or omission of extracellular Ca2+, suggesting that this isomer acts indirectly on phosphoinositide responses through activation of NMDA-type ionotropic glutamate receptors. In contrast, the responses to 1S, 3R- and 1S, 3S ACPD were unaffected by prior addition of MK-801, but were blocked by MCPG. 3. The concentration of 1S, 3R-ACPD required to half-maximally stimulate the Ins(1,4,5)P3 response (-log EC50 (M), -4.09 +/- 0.10) was significantly higher than that required to exert a similar effect on [3H]-InsPx accumulation (-log EC50 (M), -4.87 +/- 0.07; P < 0.01; n = 4). A similar marked 8-9 fold discrepancy between these two values was observed for the 1S, 3S isomer, which elicited similar maximal responses to those caused by 1S, 3R-ACPD. 4. Significant differences were also observed with respect to the ability of (+/-)-MCPG (1 mM) to cause a rightward shift in the concentration-response relationships for 1S, 3R ACPD-stimulated Ins(1,4,5)P3 (5.59 +/- 0.24 fold shift) and [3H]-InsPx (3.04 +/- 0.34 fold shift; P < 0.01; n = 4) responses, giving rise to Kd values of 218 and 490 microM for (+/-)-MCPG antagonism of the respective responses. 5. The potency difference between the 1S, 3R-ACPD-stimulated Ins(1,4,5)P3 and [3H]-InsPx responses was reduced when experiments were performed in nominally calcium-free medium ([Ca2+]e = 2 - 5 microM) and EC50 values were almost identical when extracellular calcium was reduced further by EGTA addition ([Ca2+]e < or = 100 nM). Similarly, the Kd value for (+/-)-MCPG antagonism of the 1S, 3R-ACPD stimulated [3H]-InsPx response decreased under [Ca2+]e-free conditions, approaching those obtained for the 1S, 3R-ACPD-stimulated Ins(1,4,5)P3 response in the presence of normal [Ca2+]e. 6. These data suggest that estimates of the activities of mGluR agonists and antagonists, derived by measuring phosphoinositide turnover, can differ significantly depending on whether Ins(1,4,5)P3 mass or [3H]-InsPx responses are measured. In particular, the possibility that the mGluR-mediated [3H]-InsPx response may not simply reflect direct receptor/G protein/phosphoinositidase C (PIC) activation, but may also be the consequence of stimulation of a facilitatory Ca2+-influx pathway is discussed. PMID- 8732285 TI - Antiarrhythmic action of rilmenidine on adrenaline-induced arrhythmia via central imidazoline receptors in halothane-anaesthetized dogs. AB - 1. To elucidate the role of central imidazoline receptors in the genesis of adrenaline-induced arrhythmias under halothane anaesthesia, we investigated the effects of rilmenidine, a selective agonist at imidazoline receptors, on this type of arrhythmia in dogs. Rilmenidine (1, 3, 10 micrograms kg-1, i.v.) did not affect basal haemodynamic parameters (heart rate and blood pressure), but dose dependently inhibited adrenaline-induced arrhythmias under halothane anaesthesia. 2. Although, rilmenidine has a weak affinity for alpha(2)-adrenoceptors, pretreatment with idazoxan (10 micrograms kg-1, intracisternally i.c.), an imidazoline receptor antagonist which has also alpha(2)-adrenoceptor blocking potency, blocked the antiarrhythmic effect of rilmenidine (10 micrograms kg-1, i.v.). In contrast, pretreatment with rauwolscine (20 micrograms kg-1, i.c.), a classical alpha(2)-adrenoceptor antagonist with little affinity for imidazoline receptors, did not affect the effect of rilmenidine (10 micrograms kg-1, i.v.). Furthermore, bilateral vagotomy completely blocked the antiarrhythmic action of rilmenidine (10 micrograms kg-1, i.v.). 3. It is suggested that the antiarrhythmic action of rilmenidine is due to the activation of central imidazoline receptors and that vagal tone is critical for this action of rilmenidine. PMID- 8732286 TI - Block of pancreatic ATP-sensitive K+ channels and insulinotrophic action by the antiarrhythmic agent, cibenzoline. AB - 1. We investigated the effect of cibenzoline (a class Ia antiarrhythmic drug) on basal insulin secretory activity of rat pancreatic islets and ATP-sensitive K+ channels (KATP) in single pancreatic beta cells of the same species, using radioimmunoassay and patch clamp techniques. 2. Micromolar cibenzoline had a dose dependent insulinotrophic action with an EC50 of 94.2 +/- 46.4 microM. The compound inhibited the activity of the KATP channel recorded from a single beta cell in a concentration-dependent manner. The IC50 was 0.4 microM in the inside out mode and 5.2 microM in the cell-attached mode, at pH 7.4. 3. In the cell attached mode, alkalinization of extracellular solution increased the inhibitory action of cibenzoline and the IC50 was reduced from 26.8 microM at pH 6.2 to 0.9 microM at pH 8.4. On the other hand, the action of cibenzoline in the excised inside-out mode was acute in onset with a small IC50, indicating that the drug attains its binding site from the cytoplasmic side of the cell membrane. 4. In the inside-out mode, micromolar ADP reactivated the cibenzoline-blocked KATP channels in a manner similar to that by which ADP restored ATP-dependent block of the channel. 5. The binding of [3H]-glibenclamide to pancreatic islets was inhibited by glibenclamide but not by cibenzoline. In contrast, the [3H] cibenzoline binding was displaced by unlabelled cibenzoline but not by glibenclamide. It is concluded that cibenzoline blocks pancreatic KATP channels via a binding site distinct from the sulphonylurea receptor. PMID- 8732287 TI - Protection of the aged substantia nigra of the rat against oxidative damage by ( )-deprenyl. AB - 1. We have studied the effect of (-)-deprenyl on the oxidative damage that the rat substantia nigra suffers during aging. 2. (-)-Deprenyl (2 mg kg-1, three times a week) administered for two months, beginning at 22 months of age, produced a significant increase in tyrosine hydroxylase (TH) activity (2.67 +/- 0.40 and 3.64 +/- 0.38 nmol mg-1 protein h-1 in untreated aged rats and treated aged rats respectively, P < 0.05) and in TH amount (0.072 +/- 0.012 and 0.128 +/- 0.38 absorbance 405 nm in untreated aged and treated aged rats respectively, P < 0.05). 3. The proteins of aged rat substantia nigra showed a significant decrease of carbonyl groups in treated animals compared with saline-injected control rats (136.2 +/- 21.8 and 71.5 +/- 13.2 c.p.m. microgram-1 protein in untreated aged and treated aged rats respectively, P < 0.05). 4. The carbonyl groups measured in TH enzyme showed a statistically significant decrease (42.3%) after (-)-deprenyl treatment (471.4 +/- 73.0 and 271.9 +/- 50.00 c.p.m. in untreated aged and treated aged rats respectively, P < 0.001). 5. All these results suggest that oxidative damage produced during aging is prevented by (-)-deprenyl treatment and could explain the effect of this drug in Parkinson's disease (PD) and other degenerative diseases such as Alzheimer's disease. PMID- 8732288 TI - Possible role of nitric oxide in the development of L-2-chloropropionic acid induced cerebellar granule cell necrosis. AB - 1. L-2-Chloropropionic acid (L-CPA) produces selective neuronal cell necrosis in rat cerebellum when administered orally at 750 mg kg-1 that is mediated in part through activation of N-methyl-D-aspartate (NMDA) receptors. Cerebellar granule cell death occurs between 30 and 36 h following L-CPA administration exhibiting a number of features in common with excitatory amino acid-induced cell death. We have used this in vivo model to examine the neurochemical processes following L CPA-induced activation of NMDA receptors leading to neuronal cell death in the rat cerebellum. 2. The effects of a number of compounds which potently block nitric oxide synthase in vitro were examined on L-CPA-induced neurotoxicity 48 h following L-CPA dosing, to discover whether the neuronal cell death is mediated in part by excessive nitric oxide generation. Four inhibitors were studied, NG nitro-L-arginine (L-NOARG), NG-nitro-L-arginine methyl ester (L-NAME), NG iminoethyl-L-ornithine (L-NIO) and 3-bromo-7-nitroindazole (BrNI). 3. L-NAME (50 mg kg-1, i.p. twice daily) and BrIN (50 mg kg-1, i.p. twice daily) administration prevented the L-CPA-induced loss of granule cells which can reach up to 80-90% of the total cell number in rats treated with L-CPA alone. L-NOARG (50 mg kg-1, i.p. twice daily) and L-NIO administered at either 25 or 100 mg kg-1, twice daily did not produce any significant protection against L-CPA-induced neurotoxicity. 4. Both L-NAME and BrIN also prevented the L-CPA-induced increase in cerebellar water content and sodium concentrations. L-NIO when administered at the highest doses prevented the increase in cerebellar sodium concentration but not water content. L-NIO and L-NOARG were ineffective in preventing the L-CPA-induced increases in cerebellar water and sodium concentrations. 5. L-CPA-induced reductions in cerebellar aspartate and glutamate concentrations and increases in glutamine and GABA concentrations were prevented by L-NAME and BrIn, but not by L NIO or L-NOARG. Also reductions in L-[3H]-glutamate binding to glutamate ionotrophic and metabotrophic receptors in the granule cell layer of rat cerebellum was prevented by L-NAME and BrIN, but not L-NIO or L-NOARG. 6. In conclusion, the neuroprotection offered by L-NAME and BrIN suggests that L-CPA induced cerebellar granule cell necrosis is possibly mediated by or associated with excessive generation of nitric oxide. The inability of nitric oxide synthase inhibitors, L-NOARG and L-NIO to afford protection may result from their limited penetration into the brain (L-NIO) or rapid dissociation from the enzyme. PMID- 8732289 TI - Synthesis and characterization of a selective peptide antagonist of neuropeptide Y vascular postsynaptic receptors. AB - 1. A cyclic dimeric nonapeptide neuropeptide Y (NPY) receptor antagonist, 1229U91, was synthesized by Fmoc chemistry and dimerised in solution. Its effects were assayed in mesenteric arteries from rats and mice, and in rat vas deferens. 2. Mesenteric arteries were cannulated and pressurised to 55 mmHg and the external diameters continuously measured. NPY, PYY, Leu31Pro34NPY and NPY(13-36) each caused concentration-related contractions with the order of potency PYY > or = Leu31Pro34NPY = NPY > NPY (13-36), consistent with the Y1 receptor subtype. 3. 1229U91 had no agonist activity in the arteries but caused a concentration related rightward shift of NPY (mouse arteries) or Leu31Pro34NPY (rat) concentration-response curves. The antagonism was competitive with pKBS of 7.69 +/- 0.15 and 7.47 +/- 0.13 in the mouse and rat arteries, respectively. 4. Sympathetic nerves in the vas deferens were stimulated with a single electrical field pulse every 20 s and the twitch responses recorded. NPY, PYY, Leu31Pro34NPY and NPY(13-36) inhibited the twitches with the order of potency PYY > NPY > NPY(13-36) >> Leu31Pro34NPY, consistent with the Y2 receptor subtype. 5. 1229U91 inhibited the vas deferens twitch with a shallow concentration-response curve and a time-course of inhibition distinct from that of NPY. 1229U91 (30 microM) did not cause a rightward shift of the NPY concentration-response curve. 1229U91 is at least 5 orders of magnitude less potent in the vas deferens than in rat brain Y2 binding assays reported by others, suggesting that the brain and vas deferens Y2 receptors are different. 6. It is concluded that 1229U91 is a competitive antagonist of NPY Y1 vascular receptors and has additional properties that inhibit the electrically evoked twitch of the rat vas deferens. PMID- 8732291 TI - Translocation of lipocortin (annexin) 1 to the membrane of U937 cells induced by phorbol ester, but not by dexamethasone. AB - 1. Induction of lipocortin 1 secretion by dexamethasone has been demonstrated, although the secretory mechanism is still unknown. We have studied the effects of 12-tetradecanoyl phorbol 13-acetate (TPA) and/or dexamethasone on the expression, translocation, and secretion of lipocortin 1 in U937 cells. 2. The expression of lipocortin 1 and its mRNA increased during TPA-induced differentiation of U937 cells to a maximum of 1.9 fold and 8.2 fold, respectively, after 48 h. Both the protein and the mRNA levels decreased after 48 h. 3. TPA caused the translocation of lipocortin 1 from the cytosol to the membrane of U937 cells in a time dependent manner, as determined by Western blot analysis. The translocation was concurrent with the differentiation of the cells. After 48 h of TPA treatment, 82.6 +/- 6.5% of lipocortin 1 was present in the membrane fraction compared to 41.6 +/- 1.7% in untreated cells. 4. The amount of lipocortin 1 that was externally bound (associated) with the membrane increased to 3.2 fold as the cytosol to membrane translocation of lipocortin 1 increased. 5. Dexamethasone decreased the externally bound lipocortin 1, but had no effect on the cytosol to membrane translocation. 6. This offers a model system with which the function and the secretion mechanism of lipocortin 1 can be studied. Our data is consistent with the hypothesis that the secretory mechanism is through an unknown pathway, involving the translocation of lipocortin 1 from the cytosol to the internal membranes, and then, its secretion to the external membrane. PMID- 8732290 TI - Involvement of secretory phospholipase A2 activity in the zymosan rat air pouch model of inflammation. AB - 1. In the zymosan rat air pouch model of inflammation we have assessed the time dependence of phospholipase A2 (PLA2) accumulation in the inflammatory exudates as well as cell migration, myeloperoxidase activity, prostaglandin E2 (PGE2) and leukotriene B4 (LTB4) levels. 2. A significant increase in PLA2 activity was detected in 1,200 g supernatants of exudates 8 h after injection of zymosan into rat air pouch. This event coincided with peaks in cell accumulation (mainly neutrophils) and myeloperoxidase activity in exudates and was preceded by a rise in eicosanoid levels. 3. This enzyme (without further purification) behaved as a secretory type II PLA2 with an optimum pH at 7-8 units, lack of selectivity for arachidonate release and dependence on mM calcium concentrations for maximal activity. 4. The PLA2 inhibitors manoalide and scalaradial inhibited this enzyme activity in vitro in a concentration-dependent manner. Scalaradial also inhibited zymosan stimulated myeloperoxidase release in vitro. 5. Injection of the marine PLA2 inhibitor scalaradial together with zymosan into the pouch at doses of 0.5, 1 and 5 mumol per pouch resulted in a dose-dependent inhibition of PLA2 activity in exudates collected 8 h later. Myeloperoxidase levels and cell migration were also decreased, while eicosanoid levels were not modified. 6. Colchicine administration to rats prevented infiltration and decreased PLA2 levels in the 8 h zymosan-injected air pouch. 7. These results indicate that during inflammatory response to zymosan in the rat air pouch a secretory PLA2 activity is released into the exudates. The source of this activity is mainly the neutrophil which migrates into the pouch. 8. Scalaradial exerts anti-inflammatory effects in the zymosan air pouch. PMID- 8732292 TI - Different mechanisms of Ca2(+)-handling following nicotinic acetylcholine receptor stimulation, P2U-purinoceptor stimulation and K(+)-induced depolarization in C2C12 myotubes. AB - 1. The increase in intracellular CA2+ on nicotinic acetylcholine receptor (nAChR) stimulation, P2U-purinoceptor stimulation and K(+)-induced depolarization was investigated in mouse C2C12 myotubes by use of fura-2 fluorescence to characterize the intracellular organisation of Ca2+ releasing stores and Ca(2+) entry process. 2. Stimulation of nAChRs with carbachol induced a rapid rise in internal Ca2+ (EC50 = 0.85 +/- 0.09 microM), followed by a sustained phase. The Ca2+ response evoked by carbachol (10 microM) was completely blocked by the nAChR antagonist, pancuronium (3 microM), but was not affected by the muscarinic antagonist, atropine (3 microM), or under conditions when Ca2+ entry was blocked by La3+ (50 microM) or diltiazem (10 microM). Addition of pancuronium (3 microM) during the sustained phase of the carbachol-evoked response did not affect this phase. 3. Stimulation of P2U purinoceptors with ATP (1 mM) induced a somewhat higher biphasic Ca2+ response (EC50 of the rapid phase: 8.72 +/- 0.08 microM) than with carbachol. Pretreatment with La3+ abolished the sustained phase of the ATP-induced Ca2+ response, while the response was unaffected by diltiazem or pancuronium. 4. Stimulation of the cells with high K+ (60 mM), producing the same depolarization as with carbachol (10 microM), induced a rapid monophasic Ca2+ response, insensitive to diltiazem, pancuronium or La3+. 5. Under Ca(2+)-free conditions, the sustained phase of the carbachol- and ATP-evoked responses were abolished. Pre-emptying of depolarization-sensitive stores by high K+ under Ca(2+)-free conditions did not affect the carbachol- or ATP-evoked Ca2+ mobilization and vice versa. Preincubation of the cells with ATP in the absence of extracellular Ca2+ decreased the amplitude of the subsequent carbachol-induced Ca2+ response to 11%, while in the reverse procedure the ATP-induced response was decreased to 65%. Ca2+ mobilization evoked by simultaneous addition of optimal concentrations of carbachol and ATP was increased compared to levels obtained with either agonist. 6. Preincubation with high K+ under normal conditions abolished the sustained phase of the ATP-evoked Ca2+ response. The carbachol response consisted only of the sustained phase in the presence of high K+. 7. The carbachol-induced Ca2+ response was completely abolished under low Na+/Ca(2+) free conditions, while under low Na+ conditions only a sustained Ca2+ response was observed. The ATP- and K(+)-induced responses were changed compared to Ca(2+) free conditions. 8. ATP (300 microM) induced the formation of Ins(1,4,5)P3 under Ca(2+)-free conditions with a comparable time course to that found for the rise in internal Ca2+. In contrast to ATP, carbachol (10 microM) did not affect Ins(1,4,5)P3 levels under Ca(2+)-free conditions. 9. It is concluded that the Ca2+ release from discrete stores of C2C12 myotubes is induced by stimulation of nAChRs, P2U-purinoceptors and by high K+. Only the P2U-purinoceptor and nAChR activated stores show considerable overlap in releasable Ca2+. Sustained Ca(2+) entry is activated by stimulation of nAChRs and P2U-purinoceptors via separate ion-channels, which are different from the skeletal muscle nAChR-coupled cation channel. PMID- 8732293 TI - Effect of cyclo-oxygenase inhibitors and modulators of cyclic AMP formation on lipopolysaccharide-induced neutrophil infiltration in mouse lung. AB - 1. The adult respiratory distress syndrome (ARDS) is an acute lung inflammation developed after direct or indirect contact with pathogenic agents. In the present study, a mouse model was developed to mimic this condition using aerosolized bacterial lipopolysaccharide (LPS) and to investigate the mechanisms involved in the lung inflammatory response. 2. Inhalation of LPS led to a time and dose dependent increase in tumour necrosis factor-alpha (TNF-alpha) production and neutrophil recruitment into the bronchoalveolar lavage fluid (BALF) of Balb/c mice. Under the same conditions, neutrophil infiltration was also found in the BALF of the LPS-sensitive mouse strain C3H/HeN, but was absent in the LPS resistant strain C3H/HeJ. Intranasal administration of murine recombinant TNF alpha also triggered neutrophil recruitment. 3. One hour after inhalation of LPS, half of the maximal level of TNF-alpha was measured in the BALF, but only a few neutrophils were detected at this time. The peak TNF-alpha concentration was reached at 3 h, when the neutrophil amount started to increase. At 24 h, maximal neutrophil number was found in the BALF and TNF-alpha was no longer present. 4. Pretreatment of mice under different experimental conditions demonstrated that: (a) cycloheximide almost completely blocks both neutrophil recruitment and TNF alpha production; (b) anti TNF-alpha antibodies block neutrophil recruitment; (c) indomethacin or aspirin enhance by two fold neutrophil recruitment; (d) indomethacin significantly increases TNF-alpha production 1 h after inhalation of LPS; (e) dibutyryl cyclic AMP and prostaglandin E2 (PGE2) block both neutrophil recruitment and TNF-alpha production. 5. It is concluded that aerosolized LPS in mice triggers an acute lung inflammation which can be used as a potential model of inhalational ARDS and that, strategies leading to the elevation of cyclic AMP levels in vivo can be effective in modulating LPS-induced TNF-alpha synthesis and neutrophil recruitment. PMID- 8732294 TI - Relative contribution of P2U- and P2Y-purinoceptors to endothelium-dependent vasodilatation in the golden hamster isolated mesenteric arterial bed. AB - 1. P2-purinoceptors were characterized pharmacologically in the constantly perfused isolated mesenteric arterial vascular bed of the golden hamster. Vasoconstrictor and vasodilator responses to the nucleotides ATP, ADP, 2 methylthio ATP (2MeSATP), alpha,beta-methylene ATP (alpha,beta-meATP) and uridine 5'-triphosphate (UTP) and a role for ATP in sympathetic constriction were examined. 2. At basal tone nucleotides elicited dose-dependent vasoconstriction with an observed rank order of potency of alpha,beta-meATP >> 2MeSATP > ATP = ADP > UTP (based on the doses required to elicit constrictor responses of 25 mmHg). Adenosine had no vasoconstrictor action at doses up to 5 mumol. After application of a single dose (0.5 mumol) of alpha,beta-meATP preparations were desensitized to constriction by subsequent application of nucleotides. 3. Electrical field stimulation (4-64 Hz, 90 V, 1 ms, 30 s) elicited frequency-dependent constrictions which were abolished by guanethidine (5 microM) and by prazosin (1 microM). 4. The non-selective P2-purinoceptor antagonist suramin (100 microM) did not significantly affect vasoconstrictor responses to ATP. The P2X-selective purinoceptor antagonist pyridoxalphosphate-6-azophenyl-2',4'-disulphonic acid (PPADS, 3 microM), virtually abolished responses to ATP. When the endothelium was removed vasoconstrictor responses to ATP and noradrenaline were augmented. 5. In preparations with tone raised with methoxamine (10-80 microM) nucleotides elicited vasodilatation with an observed potency order of ATP = UTP > ADP >> adenosine. 2MeSATP had relatively minor vasodilator effects and at the highest dose tested (50 nmol) elicited only vasoconstriction. alpha,beta-meATP did not elicit vasodilatation but produced further constriction of the raised tone preparation. At the highest doses of ATP and ADP (0.5 microM) responses were biphasic with vasoconstriction preceding vasodilatation. After removal of the endothelium, with the exception of adenosine, vasodilator responses to purines and to UTP were abolished; vasoconstriction to ATP, ADP, UTP and 2MeSATP was evident at the highest doses. 6. Suramin (100 microM) inhibited vasodilatation to both ATP and UTP and abolished responses to 2MeSATP. PPADS (3 microM) inhibited relaxation to 2MeSATP but did not affect relaxation to ATP, UTP, adenosine and acetylcholine and ADP. 7. Reactive blue 2 (30 microM) blocked vasodilator responses to ATP, UTP, 2MeSATP and acetylcholine; it was without effect when used at 3 microM. 8. The results of this study show that ATP elicits vasoconstriction of mesenteric arteries of the golden hamster via P2X-purinoceptors located on the smooth muscle, and vasodilatation via P2U-receptors which are located on the endothelium. 2MeSATP has marginal vasodilator activity, suggesting that P2Y purinoceptors contribute minimally to relaxation to ATP in hamster mesenteric arteries. PMID- 8732295 TI - Investigation of the involvement of the N-methyl-D-aspartate receptor macrocomplex in the development of spermine-induced CNS excitation in vivo. AB - 1. The involvement of the N-methyl-D-aspartate (NMDA) receptor macrocomplex in the development of spermine-induced CNS excitation in vivo was investigated. 2. Injection of 100 micrograms of spermine into the left lateral cerebral ventricle of female Laca mice (20-25 g) resulted in the development of two distinct phases of CNS excitatory effects which were quantified by a scoring system. 3. The first phase effects occurred within minutes of injection and generally lasted for about 1 h. Most mice showed scratching of the upper body, frequent face washing and some mice developed clonic convulsions. By about 2 h after injection, the second phase of effects began to develop in the form of body tremor which worsened with time and culminated in fatal tonic convulsions, generally within 8 h of injection. 4. Pretreatment of the mice with dizocilpine (0.3 mg kg-1, i.p.) resulted in antagonism of the first phase of spermine-induced effects, but a higher dose (0.3 mg kg-1, (x2), i.p.) was necessary to inhibit the second phase effects. 5. Whereas the glutamate antagonist, 3-((R)-2-carboxypiperazin-4-yl) propyl-1-phosphonic acid (D-CPP) (10, 20 mg kg-1, i.p.), the glycine antagonist 7 chlorokynurenate (10, 30, 50 nmol, i.c.v.), or the polyamine antagonist ifenprodil (30, 60 mg kg-1, i.p.) antagonized the first phase of effects produced by spermine, these agents given as monotherapy, were ineffective against the development of the second phase of effects. 6. Co-administration of ifenprodil with either D-CPP or 7-chlorokynurenate resulted in a dose-dependent antagonism of the development of the second phase of spermine-induced effects. 7. It is concluded that the development of the two temporally distinct phases of spermine induced effects may be mediated by pharmacologically distinct mechanisms, although the results suggest that the NMDA receptor macrocomplex may be involved in both phases of effects. Furthermore, a moderate dose of D-CPP or 7 chlorokynurenate appears to enhance the inhibitory potential of ifenprodil in vivo. PMID- 8732296 TI - Clinical gastroenterology. Introduction. PMID- 8732297 TI - Cytokines: sources, receptors and signalling. AB - Cytokines are a family of protein mediators that are important in transducing information between various cell types. These messengers are synthesized by a broad spectrum of cells. Cellular sources of cytokines include those cell types considered to play pivotal roles in the immune system as well as in inflammatory responses, including lymphocytes, monocytes and mast cells. Emerging data indicate that non-immune cells, including epithelial cells and fibroblasts, may also be important sources of certain cytokines. Cytokines fulfill a number of roles during immune and inflammatory reactions, and may display overlapping or redundant functions. In part, this redundancy may arise from the fact that cytokine receptors are not all unique entities, but may be divided into families. Many cytokine receptors have a subunit structure, with common subunits shared between receptors, and serving as affinity modifiers/signal transducers. Cytokines exert their effects on target cells by activating intracellular signalling mechanisms. In addition to 'classical' signal transduction path-ways, new data indicate that cytokines may also exemplify molecules that utilize novel signalling mechanisms, including the Jak-STAT pathways of transcriptional regulation and pathways involving the novel lipid second messenger, ceramide. In conclusion, molecular techniques have enabled the identification of many new cytokines, and the elucidation of their binding sites and mechanisms of action. This information has provided new insights into this complex area. Moreover, an understanding of the molecular basis of cytokine action and the pathways that lead to their acute and chronic effects may, in turn, facilitate interventions to prevent or modify their actions in disease states. PMID- 8732298 TI - Cytokines and gastrointestinal disease mechanisms. AB - Cytokines mediate immune responses and are detectable in the normal gastrointestinal mucosa. It is unclear how cytokines are physiologically regulated but in inflammatory enteropathies their expression is often greatly increased and may account for the tissue damage observed. T-cells may be sub divided according to the pattern of cytokines which they secrete. TH1 cytokine expression is increased in delayed type IV cell mediate immune responses whereas TH2 cytokines are raised in diseases in which humoral mechanisms are more important. Cytokines are secreted by macrophages in relatively greater amounts than from T-cells. They are non-specific products of inflammation and may account for the majority of tissue damage seen in mucosal disease. The pattern of cytokine secretion may determine the immunopathogenesis of an inflammatory disorder. The ultimate goal of cytokine research is the development of therapeutic measures based on a better understanding of their actions which may be achieved with a better understanding of the molecular immune-microenvironment in inflammatory enteropathies. Studies with transgenic mice and gene targeted mice have important implications to the understanding of the immune system and its role in intestinal diseases. PMID- 8732299 TI - Epidermal growth factor (EGF). AB - Despite the wealth of information concerning EGF and its related peptides, its precise role in the control of gastrointestinal functions is still not fully resolved. However, there is no doubt that it can have some very potent effects on the gastrointestinal tract. These may be related to the control of growth and development and to the regular control of cell renewal. Nevertheless, in the adult, EGF may only be active in response to luminal damage and repair, and furthermore this may also only occur if the luminal EGF is protected from proteolytic degradation. Notwithstanding this, 'EGF'-like responses may be evoked in the gut by intestinal TGF-alpha. The possible therapeutic use of EGF and members of its family in ulcer therapy will be discussed in later Chapters of this volume, other potential uses are in the control of necrotising enteritis and in the alleviation of the mucositis associated with cancer treatment. PMID- 8732300 TI - Transforming growth factor-alpha. AB - Major advances in understanding growth factor biology, especially in epithelial cells, have resulted from work with TGF-alpha over the past decade. It is clear that TGF-alpha is a potent epithelial oncoprotein, but equally important biological activities in normal epithelial homeostasis have been described. A number of major challenges lie ahead. Foremost is the formidable task of dissecting out the individual contributions of each EGF-related peptide in the biological response to stimulation of the EGFR. Appreciation of the complexity of heterodimerization of receptors within the EGFR family will be equally important in the final analysis. These considerations assure the continued vitality and productivity of investigation of the EGF-related peptide/EGFR axis. PMID- 8732301 TI - Transforming growth factor-beta in GI neoplasia, wound healing and immune response. AB - The last decade has been marked by tremendous advances in the biochemical and functional characterization of TGF-betas and their receptors in normal and transformed cells. TGF-betas have been shown to modulate proliferation, differentiation and motility of different cell types in a number of in vitro model systems and in some cases with some intriguing results. It is obvious that there is no simple pattern that explains the TGF-betas biological activity in vitro and their effects on cell behaviour need to be assessed in the context of an appropriate physiological cellular environment. Cell-cell and cell-matrix interactions, the differentiating status of the cell together with the functional activity of other soluble growth factors can influence how TGF-betas modulate cell behaviour. However, the overwhelming interest in this field shown by clinicians and basic scientists is rapidly increasing our understanding of how growth factors such as TGF-betas regulate the homeostasis of the GI mucosa and their role in gastrointestinal carcinogenesis. PMID- 8732302 TI - Insulin-like growth factors and inflammatory bowel disease. AB - Hallmarks of IGF-I action include synergy with other hormones and growth factors and the ability to stimulate proliferation or differentiated cell function dependent on physiological or pathophysiologial context. A complete understanding of IGF action in IBD will require analyses of mechanisms of IGF interaction with other growth factors, hormones and cytokines. GH and IGF-I may be administered to children over prolonged periods to correct growth disorders. The definition of the benefits and problems of GH/IGF-I therapy in IBD needs to distinguish between long-term and short-term effects. Short-term administration of GH and IGF-I to animal models of IBD such as the PG-PS and TNBS models, which share features of Crohn's disease (Sartor, 1992), and a recently developed murine model of ulcerative colitis induced by ingestion of dextran sulphate (Okayasu et al, 1990; Sartor, 1992; Cooper et al, 1993) could address the beneficial or detrimental consequences of short-term GH/IGF-I therapy. Adaptation of the PG-PS, TNBS and dextran sulphate models of inflammation to available transgenic mouse lines that over-express GH and IGF-I (Behringer et al, 1990; Ulshen et al, 1993), especially if over-expression is inducible, could help to define the potential benefits and problems of long-term GH/IGF-I therapy or the effects of GH/IGF-I on immune cell function and cytokine production during intestinal inflammation. It will be useful to study intestinal inflammation and complication in animal models of GH or IGF-I deficiency. In this regard, mice with targeted ablation of the IGF-I gene could be useful (Liu et al, 1993) although neonatal mortality in these models currently poses problems for in vivo studies. Development of mesenchymal cell lines from such animals could, however, provide a useful in vitro system to study the role of IGF-I in altered cell function in response to pro-inflammatory cytokines. PMID- 8732303 TI - Basic fibroblast growth factor and PDGF in GI diseases. AB - This chapter is focused on the relatively recent investigations demonstrating a pharmacological and pathophysiological role for bFGF and PDGF in ulcerative and inflammatory lesions in the upper and lower GI tract. Our initial animal model experiment revealed a potent healing of chronic cysteamine-induced duodenal ulcer in rats treated by intragastric administration of bFGF-w, the acid-resistant bFGF CS23 or PDGF-BB without decreasing gastric acid secretion or concentration. Subsequently we and others have demonstrated that these peptides accelerate the healing of chronic gastric ulcers, chronic erosive gastritis and ulcerative colitis. Contrary to the potent ulcer healing properties of bFGF and PDGF, these growth factors exert no or modest acute gastroprotection. Nevertheless, new biochemical, molecular biological and immunohistochemical studies indicate that both bFGF and PDGF play a pathophysiological role in the natural history of ulcer healing. PMID- 8732304 TI - Trefoil peptides. AB - There is a growing body of evidence supporting the hypothesis that members of the trefoil peptide family are involved actively in maintaining the integrity of the gastrointestinal mucosa and facilitating its repair. To date, three trefoil peptides are known in man: pS2, ITF and SP. Each is a secretory peptide expressed in specific compartments throughout the gut, in patterns that appear generally to be conserved between mammalian species. Ulceration, whether due to common pathological processes or experimentally induced, results in altered local expression of trefoil peptides. In diverse chronic ulcerative conditions in man, glandular structures develop within the mucosa, derived from the UACL. These UACL glands express three trefoil peptides, EGF and lysozyme, all potentially able to contribute to the healing process. In fact local goblet and endocrine cell types may also be recruited to secrete pS2 into the local environment. In experimental ulcers, in rate stomach or intestinal resection margins, there is also accentuation of trefoil peptide expression at the margins and in the poorly differentiated mucous cells extending out presumably in attempts to restore epithelial integrity. Several trefoil peptides have been expressed as 'recombinant' proteins in bacterial, baculoviral or yeast systems, and these procedures have allowed some of the biological properties of these peptides to be determined. In vitro, rITF, hITF and hSP are motogens, able to promote migration of epithelial cells. In vivo, rITF and hSP are able to prevent much of the gastric damage effect by a single dose of indomethacin, when given systemically. There is synergy between EGF and rITF both in vitro and in vivo, which may allow the development of new peptide therapies for ulceration that will maximize repair and minimize cell proliferation. PMID- 8732305 TI - Growth factors and ulcerative gastrointestinal disease. AB - A huge variety of peptides and cytokines are involved in the maintenance of mucosal integrity and in the inflammatory response at sites of ulceration. Most studies have focused on the effects of an individual factor in this complex process. However, it is becoming increasingly apparent that, to fully understand their importance in vivo, we should consider their function as part of a highly integrated system. It is also becoming clear that a relatively small number of common pathways are brought into play by the host in response to a wide variety of intestinal insults. PMID- 8732306 TI - Cytokines and inflammatory bowel disease. AB - Cytokines play an important role in the pathology of inflammatory bowel disease by determining the nature of the mucosal immune response. One way of establishing whether CD and UC are causally related to a defect in the host immune response is to look for polymorphisms that are over-represented in these populations. This is being carried out at great pace both for the cytokine genes and for some other immune response genes. A number of gene expression studies have established that those cytokines produced by activated macrophages such as IL-1, IL-6 and TNF are significantly elevated in both diseases. Differences between the two diseases are less clear, and, where they have been found, they probably reflect the accuracy and sensitivity of quantification. The picture is less clear for the T-cell derived cytokines, which are generally expressed at a lower copy number in intestinal tissue compared to the monokines. For Crohn's disease, the TH1 cytokines IL-2 and IFN may be abnormally elevated or decreased. In contrast, the TH1/TH2 profile in UC is not significantly different from normal controls. Further work is required to confirm these findings. PMID- 8732307 TI - Growth factors: potential for the management of solid epithelial tumours. AB - At present we are on the threshold of an enormous change in clinical practice. The application of molecular medicine has already started and the area of growth factor biology is particularly relevant to this endeavor (Figure 6) (Jankowski and Polak 1996). Perhaps the major limitation to this process is the rate at which the clinician can comprehend and then undertake carefully designed molecular studies in gastroenterology. In time monographs that specifically address the issue of molecular medicine in clinical gene analysis and manipulation may perhaps replace standard text books (see Jankowski and Polak, 1996). PMID- 8732308 TI - Antipsychotic agents: a survey of the prevalence, severity and burden of side effects. AB - We prospectively studied 64 male veteran out-patients with psychotic illnesses receiving one of six antipsychotic agents grouped by potency (low, mid, high, atypical) to determine: (1) prevalence of side effects; (2) cumulative side effect burden; and (3) relationship between side effects and patient-perceived burden. Patients were administered a questionnaire which included: (1) demographic information; (2) prevalence of 49 side effects; (3) visual analog scales (VAS) rating the severity of 10 selected side effects; and (4) overall side effect burden scale. Outcome measures included comparison of the prevalence, severity and overall burden of side effects among the potency groups (Part A); and association between side effect prevalence and burden, the correlation between severity and burden of certain side effects and demographic variables for the entire population (Part B). In Part A some unexpected similarities and differences were seen among data collected; few significant differences were found between groups. In Part B, 11 of 49 side effects (22%) were significantly associated with burden; the severity of 9 of 10 side effects (90%) and two demographic variables were significantly correlated with burden. These findings suggest that the prevalence and severity of many side effects of antipsychotic agents may affect a patient's perception of side effect-related burden. The side effect burden ratings suggest that the low-potency agents were the most burdensome, while the atypical group was the least burdensome. PMID- 8732309 TI - Psychological effects of a drug that facilitates brain AMPA receptors. AB - The effects of 1-(quinoxalin-6-ylcarbonyl)piperidine (CX516), a centrally active compound that facilitates AMPA receptor-mediated synaptic responses, were tested in human subjects. Separate tests of delayed recall were given prior to and nearly 3 h after administration of placebo (n = 12) or drug (n = 36). Control subjects exhibited poorer performance in the second session than in the first while subjects given 600-1200 mg of the drug did not. There were no pre- vs post treatment differences in immediate recall in either group. The drug did not reliably affect self-assessment scores for any of several psychological variables but did disrupt the normally present correlations for within-subject changes in the variables. These results suggest that AMPA receptor modulators may (1) improve memory under some circumstances and (2) produce psychological effects that are subtle or not related to specific mood states. PMID- 8732310 TI - Treatment of obsessive-compulsive disorder with fluvoxamine: a multicentre, double-blind, placebo-controlled trial. AB - One hundred and sixty patients with a primary diagnosis of obsessive-compulsive disorder were enrolled in a multicentre, randomized, double-blind, placebo controlled study of fluvoxamine. After a placebo washout phase, patients were randomized to treatment with placebo or fluvoxamine (100-300 mg/day) for 10 weeks. Seventy-eight patients in each group were evaluable for efficacy. Fluvoxamine was significantly more effective than placebo as assessed by the Yale Brown Obsessive-Compulsive Scale (Y-BOCS), the National Institute of Mental Health Obsessive-Compulsive (NIMH-OC) scale and the Global Improvement item of the Clinical Global Impression (CGI) scale. The percentage of patients classified as "responders" (much or very much improved according to the Global Improvement item) was also significantly higher in the fluvoxamine group from Week 6 onwards, with 33.3% of fluvoxamine-treated patients and 9.0% of those given placebo classified as "responders" at endpoint. The "responders" to fluvoxamine experienced a substantial clinical benefit as reflected in decreases in their Y BOCS and NIMH-OC scores. Fluvoxamine was well tolerated with the majority of adverse events considered mild or moderate. PMID- 8732311 TI - Comparison of the effects of astemizole/pseudoephedrine and triprolidine/pseudoephedrine on CNS activity and psychomotor function. AB - The sedative properties of astemizole-D and triprolidine-D were compared in a double-blind, placebo-controlled, repeated-measures design study comprising three experimental treatments, each with a duration of 2 days (n = 12). Sedation was assessed by continuous electroencephalographic measurement (C-EEG), intermittent performance testing and subjective measures. C-EEG monitoring revealed that triprolidine-D produced significantly more daytime sedation and drowsiness than either astemizole-D or placebo (p < 0.05). Intermittent performance testing did not reveal consistent psychomotor deficits. There were no differences from placebo; the only significant findings showed that astemizole-D improved tracking accuracy at T + 65 h (p < 0.05) compared to baseline. Also, when scores were summed across all time points, astemizole-D improved scores significantly in contrast to triprolidine-D for the total scores (p < 0.05). It is concluded that, in contrast to triprolidine-D, astemizole-D does not produce daytime drowsiness or sedation. PMID- 8732312 TI - Sequential administration of augmentation strategies in treatment-resistant obsessive-compulsive disorder: preliminary findings. AB - Given that an important proportion of patients with obsessive-compulsive disorder (OCD) fail to respond adequately to serotonin (5-HT) reuptake inhibitors (SRI), augmentation strategies aimed at enhancing further 5-HT transmission by different mechanisms were attempted sequentially in 13 SRI-resistant patients. Addition of the 5-HT1A l beta-adrenergic antagonist pindolol did not alter OCD symptomatology but produced a rapid improvement of depressive symptoms. The 5-HT1A agonist buspirone as well as 5-hydroxytryptophan, the immediate precursor of 5-HT, added to the SRI-pindolol regimen, were not effective in attenuating the intensity of OCD. Tryptophan, added to the SRI-pindolol regimen, produced a significant improvement after 4 weeks, with further amelioration after 6 weeks (36% decrease of the Yale-Brown Obsessive Compulsive Score), which was maintained with treatment prolongation. PMID- 8732313 TI - Quality of life in depressed patients: comparison of fluoxetine and major tricyclic antidepressants. AB - Quality of life is one of the effectiveness measures used to assess the impact of medical interventions. This paper describes results of a study on quality of life of depressed patients treated with fluoxetine or tricyclic antidepressants (TCAs) such as amitriptyline and clomipramine. The quality of life of patients was measured using the Short Form (SF-36), a widely accepted and validated questionnaire. Depressed patients (n = 845) were observed using a cross-sectional design. Patients who had been under treatment for at least 1 week (amitriptyline, clomipramine or fluoxetine) and met the DSM-III-R criteria for major depressive disorder were included. Similar sociodemographic profiles were found across treatment groups. However, more patients with a history of depression were found in the TCA vs the fluoxetine group. Compliance was dramatically lower and length of treatment higher for TCA-treated groups. Controlling for confounding variables, the TCA-treated group also scored lower than the fluoxetine-treated one for general health perception, and social and physical functioning. These differences are not explained by symptom severity since Hamilton scores were similar across treatment groups. These results suggest that fluoxetine treatment may be associated with higher levels of social functioning and health perception than usual TCA treatment. PMID- 8732314 TI - Risperidone appears safe in patients with antipsychotic-induced blood dyscrasias. AB - Two cases are reported who developed blood dyscrasias, involving neutropenia and thrombocytopenia respectively, after treatment with phenothiazine and butyrophenone antipsychotics. A subsequent good therapeutic response was obtained with risperidone. It is suggested that risperidone may be considered as an alternative when blood dyscrasias occur with classic antipsychotics. PMID- 8732315 TI - Carbamazepine augmentation therapy in three patients with trazodone-resistant unipolar depression. AB - In three patients with trazodone-resistant unipolar depression, carbamazepine 400 mg/day was co-administered for 4 weeks. They all showed favourable responses; two non-delusional patients completely or almost recovered, while one delusional patient showed considerable improvement in depressive symptoms other than delusions of guilt. Plasma concentrations of trazodone and its active metabolite m-chlorophenylpiperazine (m-CPP) were decreased, while the m-CPP/trazodone ratio was increased in all cases. The present report suggests that carbamazepine augments antidepressant effects of trazodone by an enhancement of serotonin function. PMID- 8732316 TI - Sertraline withdrawal in two brothers: a case report. AB - Two case histories are reported describing sertraline withdrawal in two brothers. The case histories document withdrawal symptoms of (1) dysequilibrium (2) dysesthesias (3) dizziness and (4) a flushing sensation. The literature of selected serotonin reuptake inhibitors (SSRI) withdrawal is reviewed. Advocated is the monitoring of patients' withdrawal symptoms from SSRI's even when prescribed at modest doses. PMID- 8732317 TI - Trihexyphenidyl as a possible therapeutic option in clozapine-induced nocturnal enuresis. AB - Enuresis is an adverse event of clozapine treatment. The occurrence of nocturnal functional enuresis in two schizophrenic patients during the initial phase of clozapine therapy is reported. Beneficial effect of trihexyphenidyl administration (5 mg at 21.00 h) on clozapine-induced enuresis is clearly demonstrated in one patient. Trihexyphenidyl discontinuation and subsequent readministration in this patient led to corresponding recurrence and then disappearance of enuresis. Involvement of the cholinergic system has been proposed as one of the possible pathophysiological mechanisms of clozapine induced enuresis. PMID- 8732318 TI - The use of risperidone in treatment-resistant schizophrenia: two case reports. AB - The role of risperidone in the management of treatment-resistant schizophrenia remains unclear. We describe two patients with treatment-resistant schizophrenia characterized by unremitting delusions, thought disorder and self-neglect who had a sustained improvement in their symptoms soon after starting risperidone. Risperidone may be a suitable alternative to clozapine in some patients unresponsive to conventional antipsychotics. PMID- 8732319 TI - Increase of aldosterone secretion following acute haloperidol administration: possible clinical implications. AB - The present study was designed to investigate the effect of haloperidol on plasma corticosteroid levels in a small sample of unmediated psychiatric patients requiring acute care. Seven young male normotensive subjects in metabolic balance received a single dose of haloperidol (2 mg i.v.). Blood samples were collected for the radioimmunoassay of plasma renin activity, cortisol and aldosterone concentration at baseline and 3, 6, 12 and 24 h after injection. In five out of seven patients a significant, transient elevation of plasma aldosterone level was observed within 12 h from administration. In contrast, plasma renin activity and cortisol concentration were unchanged. Possible clinical implications of the neuroleptic-associated aldosterone elevations are discussed. PMID- 8732320 TI - Scientific basis of health services. PMID- 8732321 TI - Computerized assessment of common mental disorders in primary care: effect on clinical outcome. AB - OBJECTIVE: A randomized controlled trial was conducted to examine the clinical effectiveness of providing general practitioners (GPs) with the results of a self administered computerized assessment of common mental disorders. METHOD: Attenders at a general practice in a deprived inner city area of South London were identified using case finding questionnaires. Six hundred and eighty-one subjects were randomly allocated to three groups which differed in the information provided to the GP: 1) no additional information was given to the GP; 2) the results of the 12 item General Health Questionnaire (GHQ) were given to the GP (the GHQ is a paper and pencil questionnaire that assesses common mental disorders); 3) the results of a self-administered computerized assessment (PROQSY) of common mental disorders were provided for the GP. RESULTS: Clinical outcome was assessed using the 12-item GHQ. Consultations with the GP, prescriptions and referrals within and outside the practice were also recorded. The group in whom the GP received the results of the computerized assessment showed a modest clinical improvement, relative to the other two groups after 6 weeks. There was no difference in clinical outcome between the groups at 6 months. There appeared to be no increase in consultations or prescriptions in the computerized assessment group. CONCLUSIONS: Self-administered computerized assessments for psychiatric disorder have potential as a means of improving the clinical outcome of patients in primary care. It is likely that the effectiveness of the approach would be greatly increased by linking the results of computerized assessments to clinical practice guidelines, tailored to the individual patient by means of computerized technology. PMID- 8732322 TI - "I know what to do, but it's not possible to do it"--general practitioners' perceptions of their ability to detect psychological distress. AB - BACKGROUND: Accurate detection of psychological distress in patients is a prerequisite of specific diagnosis and active management. Studies have shown that improved detection is related to altered management and to improved patient outcomes: there may also be a link with improved patient satisfaction. OBJECTIVE: Many factors in the doctor, patient, and context of the consultation may influence whether or not a GP identifies psychological distress in a patient; whatever the triggers to detection, it has been shown that specific training in appropriate skills can alter clinician behaviour and improve detection rate. This study examined the GPs' own perceptions of the influences on their performance as detectors of psychological distress. METHOD: A postal questionnaire yielded nineteen GPs who were personally interviewed for the study. RESULTS AND CONCLUSIONS: The study highlights GPs' sense of the difference between possessing the necessary skills and employing them in daily practice. This has implications for training and clinical practice in this area. PMID- 8732323 TI - A prospective controlled trial of computerized decision support for lipid management in primary care. AB - OBJECTIVES: This study aimed to assess the uptake and effect in primary care of a computerized decision support system (DSS) for the management of hyperlipidaemia. METHOD: A prospective controlled trial was conducted in 25 practices covering a population of 150,000 in the city of Birmingham. The Primed system, a specialist developed, rule based DSS for general practice, was introduced prospectively after a 3-month baseline data collection. The main outcome measures were nine months' data on prescribing of lipid lowering agents; use of laboratory tests; and referrals to secondary care for the investigation of hyperlipidaemia. RESULTS: System use was lower than expected. A shift was observed towards requests for appropriate follow-up of previously abnormal lipid results and a greater emphasis on full lipid profiles, in line with the DSS guidelines. Referrals showed a 55% decrease on those expected (NS). The prescribing evaluation revealed a large variation between practices, but no significant alteration following system use. Views of users favoured decision support as a concept, but criticised technical problems with the system. CONCLUSIONS: Greater integration of DSS software and practice based data handling systems is needed. The mode of data capture, and hence both the content and form of knowledge representation, in DSS must take greater account of the primary care consultation process if such systems are to be of use to practitioners. PMID- 8732325 TI - Evaluation of preventive and health promotion activities in 166 primary care practices in Spain. The Coordinating Group For Prevention and Health Promotion in Primary Care in Spain. AB - BACKGROUND: Since January 1989 the Spanish Society of Family and Community Medicine has supported the Program of Prevention and Health Promotion (PAPPS) with the following objectives: to detect possible difficulties in the implementation of recommendations especially among asymptomatic low-risk adults and children in Spain, to disseminate those recommendations, and to encourage research in prevention and health promotion in primary care. OBJECTIVE: We wished to evaluate the implementation of PAPPS in primary care practices. METHODS: A retrospective audit of medical records in practices enrolled in the programme was carried out. From 166 primary care practices widely distributed in Spain 21,631 patients were selected using systematic sampling. RESULTS: Age was positively associated with correct performance in the adult population, while in the paediatric population it was negatively associated. Females had higher probability than males of having blood pressure measurement (OR 1.34, 95% CI 1.24 1.45) and smoking counselling (OR 1.38, 95% CI 1.33-1.43) correctly performed. Practices not using the preventive chart had a lower probability than user practices of correctly performing blood pressure measurement (OR 0.69, 95% CI 0.62-0.76), alcohol counselling (OR 0.66, 95% CI 0.60-0.73) and smoking counselling (OR 0.69, 95% CI 0.63-0.76). Non-teaching practices had a higher probability than teaching centres of correct performance of blood pressure measurement (OR 1.47, 95% CI 1.35 to 1.59), alcohol counselling (OR 1.67, 95% CI 1.54 to 1.79) and smoking counseling (OR 1.39, 95% CI 1.29 to 1.51). CONCLUSIONS: There is an unequal level of performance depending on the procedure and on the target population. A preventive chart might be useful in improving implementation of periodic health maintenance. Teaching centres with training physicians need to put more emphasis on prevention and health promotion activities. PMID- 8732324 TI - Antihypertensive therapy in older patients with isolated systolic hypertension: the Syst-Eur experience in general practice. The Syst-Eur Investigators. AB - BACKGROUND AND OBJECTIVE: This interim report from the Syst-Eur trial investigated the level of blood pressure control achieved during the double-blind period in patients followed in general practices. METHODS: In the Syst-Eur trial elderly patients (60 years or older) with isolated systolic hypertension were randomized to either active or placebo treatment. Active treatment consisted of nitrendipine combined with enalapril and/or hydrochlorothiazide to reduce systolic pressure to < 150 mmHg and by > or = 20 mmHg. Matching placebos were used in the control group. RESULTS: This analysis was restricted to patients of general practitioners who had been followed for at least 12 months. The placebo (N = 204) and active treatment (N = 217) groups had similar characteristics at randomization. At one year, the difference in sitting pressure between the two treatment groups was 10 mmHg systolic and 4 mmHg diastolic. Fewer patients remained on monotherapy in the placebo than in the active treatment group and on placebo the second and third line medications were started earlier. Nitrendipine tablets were discontinued in 10 patients on placebo and in 21 patients assigned to active treatment (P < 0.001 for all comparisons). CONCLUSIONS: A significant blood pressure reduction can be achieved and maintained in older patients with isolated systolic hypertension followed by general practitioners. Whether this blood pressure reduction results in a clinically meaningful decrease of cardiovascular complications is under investigation. PMID- 8732326 TI - Can we understand partnerships in general practice? A pilot study. AB - BACKGROUND: Partnerships have been investigated in different professions, but other than identifying problems, little work has been carried out on general practice. OBJECTIVE: The aim of this present study was to develop methods for studying partnerships in general practice. METHOD: A tripartite methodological approach was used, with questionnaires adapted from other instruments in use in other professions, followed by an individual interview with each partner, and non participant observation at a partnership meeting. Results for one case-study partnership are given. RESULTS: There were no major differences between the partners on all dimensions measured; the minor differences indicated by the results of the questionnaires were corroborated by the partner interviews and observations. CONCLUSIONS: We conclude that the use of such techniques could provide support to partnerships going through significant periods of change. PMID- 8732327 TI - Constipation: a different entity for patients and doctors. AB - BACKGROUND: Medical definitions of constipation vary. Some are specific, based on the interval between defecations, with three days or twice weekly as the upper limit of normality. Others are vague. OBJECTIVE: This study investigates patients definitions, attitudes and management of 'constipation' and compares their definitions with physicians' criteria. METHODS: Survey of 1) 531 randomized patients of family clinics of the sick Fund of the General Federation of Labor of Israel, and 2) 100 randomized specialists and residents in family medicine. RESULTS: The present survey showed that constipation is more common in women and in the older (> 40 years) age group. The most important finding of this investigation was the wide discrepancy in the criteria used by doctors and patients to diagnose constipation. Fifty per cent of the patients defined constipation differently from accepted medical definitions--27% of the patients defined it as defecation every 2 days or less and 25% as hard stool alone. All 57 doctors who were asked the same question defined constipation as defecation every 3 or 4 days or less, sometimes in combination with hard stool. CONCLUSIONS: This study suggests that patients and doctors refer to different entities when they talk of 'constipation'. It is the doctor's responsibility to ensure that this misunderstanding is avoided so that unnecessary tests and treatment are not undertaken and, patient-doctor acrimony is avoided. PMID- 8732328 TI - Colorectal cancer and polyps in patients aged 40 years and over who consult a GP with rectal bleeding. AB - BACKGROUND: Rectal bleeding is common in the community and in general practice, but few studies have examined the causes of rectal bleeding in patients presenting to general practitioners. OBJECTIVE: To determine the frequency of neoplastic conditions in patients with rectal bleeding presenting in general practice and to explain the associations between presenting symptoms and final diagnoses. METHODS: We conducted two studies, the first in 1989, the second in 1991, in which we invited Danish general practitioners to register 3-4 patients aged 40 and over presenting with rectal bleeding. RESULTS: In Study 1 among 208 patients aged 40 and over and presenting with a first episode of rectal bleeding, colorectal cancer and polyps were present in 15.4 and 7.7%, respectively. In Study 2 among 209 patients aged 40 and over and presenting with overt rectal bleeding, 156 reported a first bleeding episode or a change in their usual bleeding pattern, and in this group colorectal cancer and polyps were diagnosed in 14.1 and 11.5%, respectively. In the group with unchanged bleeding the cancer polyp prevalence was 6.7% (P < 0.05). The patients in both studies were followed through a yearly letter to the GP for at least 32 and 22 months, respectively. CONCLUSIONS: A joint analysis of the two study populations showed that only age and change in bowel habit contributed to differentiating the cancer from the non cancer patients. PMID- 8732329 TI - Interruptions during general practice consultations--the patients' view. AB - BACKGROUND: Although most aspects of the consultation have been extensively reported there is very little information on the effects of interruptions on the consultation. OBJECTIVE: We wished to discover the patients' view of interruptions. METHODS: In this pilot study the sources and frequency of interruptions to the consultations of a single general practitioner were measured. The effects of interruptions on 102 patients whose consultations were interrupted were then ascertained using a simple questionnaire. RESULTS: The overall interruption rate was found to be 10.2%. The telephone was the commonest source of interruption, accounting for 50% of interruptions. Although most patients did not perceive the interruption as having an important effect on the consultation, 20% of patients did feel that the interruption had a bad effect on the consultation and 40% of patients felt it would have been better not to have been interrupted. A majority of patients (52%) did not feel that the reason for the interruption was important. Although most patients did not feel affected by the interruption, a significant minority (18%) of patients had a strongly negative emotional response to the interruption. CONCLUSIONS: In view of these findings the need for further work has been highlighted. PMID- 8732330 TI - Anxiety and depression in general practitioners: associations with type of practice, fundholding, gender and other personal characteristics. AB - BACKGROUND: There is evidence both that a doctor's own well-being is closely associated with efficiency and positive attitude to patients, and that levels of stress, anxiety and depression in doctors are rising. OBJECTIVES: This postal survey aimed to measure anxiety and depression levels in general practitioners in 1994 and identify any associations with personal and practice characteristics. METHODS: All general practitioners with patients in Staffordshire were invited to complete the Hospital Anxiety and Depression (HAD) scale. RESULTS: Six hundred and twenty of 896 general practitioners replied (response rate 69%). No gender differences were found in rates of anxiety and depression; overall, 19% of respondents were 'cases' of anxiety and 22% others had borderline anxiety scores; 10% were 'cases' of depression and 16% others had borderline depression scores. Anxiety 'caseness' was associated with living alone, amount of on-call duties undertaken, and being fourth/fifth wave fundholders. Depression 'caseness' was associated with having little free time from practice work, amount of on-call, being single handed, and working in a non-training practice. CONCLUSIONS: The authors concluded that the level of mental ill-health in general practitioners is a matter of concern and is associated with workload. PMID- 8732331 TI - Explaining variations in the frequency of night visits in general practice. AB - BACKGROUND AND OBJECTIVE: The study identifies factors which explain variations in the frequency of night visits made by general practitioners, and is based on a cross-section regression analysis of practices in Lincolnshire in 1993. METHOD: The independent variables comprise 22 general practice characteristics, related to health and socio-demographic structure of practice lists, practice management and organization, location and social deprivation. RESULTS AND CONCLUSIONS: A final model containing 6 of the 22 variables originally entered succeeds in explaining 72 per cent of the variation in the frequency of night visits per GP per quarter-year. These variables are average list size per GP, frequency of home visits, maternity claims, list inflation owing to the influx of temporary residents, the number of proximate practices and unemployment. PMID- 8732332 TI - Paracetamol prescribing--an epidemic? AB - BACKGROUND: Although we are uncertain of its therapeutic mechanism, paracetamol is seen as a safe drug, especially for children. However, adult fatalities from overdose and its association with hepatotoxicity have cast doubt on its safety. OBJECTIVE: We aimed to establish the prescribing patterns of paracetamol in the United Arab Emirates (UAE). METHOD: The prescribing patterns in the UAE for paracetamol were studied by obtaining information on national utilization and recording in detail the pattern of prescribing in one primary health care centre. RESULTS: Paracetamol was included in 35.5% of all prescriptions from the study practice. Of these, 58.5% were for children under 12 years of age and overall 13.5% were for those infants under one year of age. CONCLUSIONS: National prescribing utilization is in keeping with the prescribing patterns of the index practice. Paracetamol prescribing is reaching epidemic proportions and the potential dangers of hepatotoxicity and the inhibition of the immune response in children are discussed. PMID- 8732333 TI - Secondary prevention of excessive alcohol use: assessing the prospects of implementation. AB - BACKGROUND: Alcohol risk and harm reduction is a public health approach that goes beyond specialized treatments for alcoholism. The greatest potential for reducing alcohol risk and harm in a population depends on the extent to which health care practitioners use secondary prevention programmes. OBJECTIVE: We aim to assess the factors that affect the prospects of disseminating comprehensive, secondary prevention programmes into mainstream practice. METHOD: A decision balance was used to assess the prospects of practitioners implementing comprehensive programmes systematically. The stages-of-change model provides perspectives about behaviour change with regard to patients, practitioners and practice settings. RESULTS AND CONCLUSIONS: Programme implementation is extremely unlikely given the current organization of health care settings. To maintain the use of such programmes, we need to change the "unit of leverage" in the system: from the clinical encounter--that is, practitioners working with individual patients in a case-finding manner--to an organizational level--that is, the appropriate use of managerial and information systems supporting health care settings to identify at risk patients systematically as they enter primary care and hospital settings. With appropriate infrastructure support, practitioners will be able to fulfil the potential for as well as maintain the use of comprehensive, secondary prevention programmes to reduce alcohol risk and harm in the population. PMID- 8732334 TI - Selections from current literature: abortion, breast cancer and causal inferences. PMID- 8732335 TI - Research and history. PMID- 8732336 TI - Shared care and HIV: is there more to say? PMID- 8732337 TI - 'Undifferentiated progenitor cells' in focal nodular hyperplasia of the liver. AB - Focal nodular hyperplasia is a tumour like lesion, characterized by a central fibrous scar with irradiating fibrous septa that surround hyperplastic nodules and contain multiple bile ductules. The origin of the bile ductular structures is not clear. Recently, we found evidence for the existence of human counterparts of rat oval cells (potential stem cells) that have the ability of differentiating towards both bile duct cells and hepatocytes. These cells were found in regenerating human liver as well as in chronic cholestatic conditions. Because cholestatic features are seen in focal nodular hyperplasia, we initiated an immunohistochemical study on 23 surgical specimens using antibodies specific for cytokeratins 7 and 19 (bile duct type cytokeratins), OV6 (rat oval cell marker), chromogranin-A (shown to be positive in reactive bile ductules and human oval like cells) and neural cell adhesion molecule--NCAM (shown to be positive in reactive bile ductules) to investigate whether 'undifferentiated progenitor cells' are also present in focal nodular hyperplasia. Electronmicroscopy was applied in five cases. Bile ductules invariably showed immunoreactivity for CK7 and 19, OV6, chromogranin-A and NCAM. In addition, small individual cells with an oval nucleus and a small rim of cytoplasm, in the vicinity of the septa, were immunoreactive for chromogranin-A, CK7 and 19 and OV6. These cells were hardly recognizable on routine light microscopy. Clusters of periseptal hepatocytes, seemingly in continuity with bile ductular structures, had a transitional phenotype: they stained positive for chromogranin-A, CK7 and OV6 and sometimes formed liver cell rosettes. The number of OV6-positive hepatocytes was greater than the number of chromogranin-A and CK7 positive hepatocytes. This indicates that, in human liver, OV-6 is not purely a marker of progenitor cells. Ultrastructurally, small immature cells, highly resembling rat oval cells, were recognized in the vicinity of septa. In addition, transitional cells displaying characteristics both of hepatocytes and bile duct cells were also present. These results confirm the presence of 'undifferentiated progenitor cells' in focal nodular hyperplasia and suggest that the ductular reaction in these lesions results, at least partly, from activation of these cells. PMID- 8732338 TI - Adhesion molecule expression and leucocyte trafficking following immunotherapy with recombinant interleukin-2. AB - The relevant anti-tumour mechanisms of recombinant interleukin-2 (rIL-2) in vivo are unclear but an influx of T-lymphocytes and macrophages has been noted in regressing lesions. One of the dose limiting toxicities of rIL-2 is the development of a capillary leak syndrome attributed to widespread endothelial activation. Changes in expression of endothelial and leucocyte-associated adhesion molecules were assessed in tumour and uninvolved skin in patients with metastatic breast cancer receiving rIL-2. Increased expression of intracellular adhesion molecule-1, its leucocyte-associated ligand, leucocyte function associated molecule-1, vascular cell adhesion molecule and its ligand, very late after activation antigen-4 as well as members of the selectin family of adhesion molecules, were noted in uninvolved skin following rIL-2. Expression of these adhesion molecules was noted in tumour stroma before rIL-2 but little change was observed following rIL-2 infusion. An influx of monocytes and T-lymphocytes (expressing the IL-2 receptor and of the memory subtype) and a lower number of neutrophils was noted in uninvolved skin following rIL-2. Although monocytes and T-lymphocytes were present in tumour stroma before rIL-2 no changes were observed following infusion. The changes noted in the dermis contrast with those seen at tumour sites and may partly explain the low therapeutic index of rIL-2. PMID- 8732339 TI - Clear cell malignant myoepithelioma of the salivary glands. AB - Previous reports of monomorphic clear cell carcinoma of the salivary glands have shown inconsistent results with immunohistochemistry, especially for S-100 protein, and this has led to uncertainty about the nature of these tumours. We believe that much can be explained by considering this group as comprising not one but two separate neoplasms, one epithelial and the other myoepithelial. The former has been described as hyalinizing clear cell carcinoma--it generally occurs in the minor salivary glands, and strongly expresses cytokeratins but not S-100 protein or alpha smooth muscle actin. In contrast, this study presents five primary malignant tumours of the major salivary glands also composed largely of a single population of clear cells, but displaying histological and immunohistochemical features of myoepithelial differentiation, such as the formation of collagenous spherules and expression of S-100 protein and actin. A small number of similar tumours have been reported previously. We, therefore, believe that these neoplasms represent a clear cell variant of malignant myoepithelioma (myoepithelial carcinoma). PMID- 8732340 TI - Down regulation of bcl-2 by p53 in nasopharyngeal carcinoma and lack of detection of its specific t(14;18) chromosomal translocation in fixed tissues. AB - High levels of bcl-2 protein have been found in a wide variety of human cancers. Since p53 gene inactivation occurs in over half of human cancers, it is possible that loss of p53-mediated repression of bcl-2 gene expression accounts, at least in part, for the frequent abnormalities in bcl-2 protein production seen in tumours. By using immunohistochemical methods, we have analysed thirty-three nasopharyngeal carcinomas for p53 and bcl-2 expression. We found an inverse correlation between the expression of these two proteins (P < 0.001). Moreover, we utilized universal oligonucleotide primers of a region 5' to the bcl-2 MBR and at the 3' end of JH segments to initiate a DNA polymerase chain reaction that amplified these bcl-2-JH junctures. Of the twelve nasopharyngeal carcinomas expressing bcl-2, none showed a t(14;18) chromosome translocation. These findings may indicate potential mechanisms by which bcl-2 regulates apoptosis. PMID- 8732341 TI - The demonstration of pericryptal fibroblasts in background mucosa and dysplasia complicating ulcerative colitis. AB - The demonstration of pericryptal fibroblasts in background mucosa and dysplasia in ulcerative colitis was investigated by immunohistochemistry using monoclonal antibody for alpha-smooth muscle actin. Pericryptal fibroblasts were reduced in 18 (26%) of the 68 sections of non-dysplastic mucosa. The reduction was significantly correlated with goblet cell depletion and villous change. Pericryptal fibroblasts were more frequently reduced (50%) in dysplastic mucosa, the reduction being greater in villous than in non-villous dysplasia. Pericryptal fibroblast development was not related to grade of dysplasia (low or high-grade), distance from carcinoma (adjacent to or distant from carcinoma) or growth pattern (polypoid or non-polypoid). These findings suggest that: 1 reduction of pericryptal fibroblasts in background mucosa may relate to the development of dysplasia in ulcerative colitis, 2 reduction of pericryptal fibroblasts in villous regeneration, analogous to that in dysplasia, reinforces the hypothesis that villous change may be a marker for risk of development of carcinoma in ulcerative colitis. PMID- 8732342 TI - Rhabdoid tumour of the kidney: a clinicopathological study of 22 patients from the International Society of Paediatric Oncology (SIOP) nephroblastoma file. AB - We present 22 (0.9%) cases of rhabdoid tumour of the kidney diagnosed amongst 2392 renal tumours in children. The patients ages ranged from 3 weeks to 94 months (median 7 months) and the female:male ration was 1.2:1. Clinically, they presented with an abdominal mass but four (18%) children also had hypercalcaemia and one (4.5%) developed a brain tumour (primitive neuroectodermal tumour). None of the children presented with stage I disease, five (23%) had stage II, ten (46%) stage III, and five (23%) stage IV disease. Two (9%) patients had bilateral tumours. Histologically, the vast majority (20/22) of the tumours exhibited a classical pattern but other histological patterns were also noted. Immunohistochemical studies performed in 12 cases showed vimentin positivity in all cases, CAM 5.2 in eight, epithelial membrane antigen in six, neuron specific enolase in four, S-100 protein in eight, and desmin in one case. In only 12 of the 22 tumours was there agreement between the reporting pathologist and the panel on a diagnosis of rhabdoid tumour of the kidney. Eight tumours originally diagnosed as rhabdoid tumour of the kidney were found to be other renal tumours and in another ten cases the initial diagnosis was changed by the panel to rhabdoid tumour. Metastases developed in 18 (82%) of the children--in eight they were present at the time of diagnosis and in 10 they developed from 2 weeks to 9 months after initial diagnosis. Metastases were found in the lung, abdomen, lymph nodes, liver, bone and brain. Of the 19 children with adequate follow-up, only two (10.5%) with stage II disease are alive, while 17 (89.5%) died 2 weeks to 20 months after the diagnosis. PMID- 8732343 TI - Follicular dendritic cells in bone marrow lymphoproliferative diseases: an immunohistochemical study including a new paraffin-resistant monoclonal antibody, DR53. AB - Two-hundred and twenty-one bone marrow biopsies with lymphoid infiltrates were studied histologically and immunohistochemically, to assess the incidence and the pattern of follicular dendritic cells. Three monoclonal antibodies selective for follicular dendritic cells were used: CD21, CD35 and DR53, all reactive on paraffin-embedded material. Follicular dendritic cells were present in two of 38 benign lymphoid aggregates, 92 of 134 low grade B-cell lymphomas (45 of 62 lymphocytic, 16 of 27 lymphoplasmacytoid, 0 of six hairy cell leukaemias, five of six centrocytic, 19 of 21 centroblastic-centrocytic, seven of 12 low grade NOS), one of 23 high grade B-cell lymphomas, 0 of 10 T-cell lymphomas, 0 of three Hodgkin's disease and four of 13 suspicious infiltrates. Follicular dendritic cells were found in lymphomatous involvement with nodular, patchy and massive growth pattern, but not in interstitial ones. They formed follicle-like networks, whose number and size were directly correlated to the tumour mass. The origin and frequency distribution of follicular dendritic cells in bone marrow biopsy lymphomas is discussed and the diagnostic relevance of follicular dendritic cell immunostaining in routine bone marrow biopsy lymphoid infiltrates is assessed. PMID- 8732344 TI - Intermediate filament aggregates in mitoses of primary cutaneous neuroendocrine (Merkel cell) carcinoma. AB - Primary cutaneous neuroendocrine carcinomas express different kinds of intermediate filaments and frequently in a 'paranuclear globular' pattern. We have observed the same pattern not only in interphase but also in mitotic cells, which are very frequent in these tumours. We report a quantitative and morphological study of eight primary cutaneous neuroendocrine carcinomas stained with different antibodies against cytokeratins (CAM 5.2 and anticytokeratin 20), neurofilaments (70 kDa and 200 kDa) and peripherin. We have found a predominance of CAM 5.2 expression in interphase cells and of neurofilament proteins in mitotic cells; 87.02% of the interphase cells were positive with CAM 5.2 whereas only 6.08% were positive for neurofilaments (P < 0.01); 35.41% of the mitotic cells were positive with CAM 5.2, whereas 50% were positive for neurofilaments (P < 0.01). A correlation between a globular pattern of intermediate filament proteins and prognosis has not been found. We describe for the first time the division of neoplastic cells with a globular pattern; the presence of intermediate filament proteins with a globular pattern in all mitotic stages; and the uneven distribution of this formation between the two daughter cells. PMID- 8732345 TI - Hyalinizing trabecular carcinoma of thyroid gland. AB - We describe two cases of encapsulated thyroid tumours which displayed the classic morphological features of hyalinizing trabecular adenoma. In addition, both were characterized by focal invasion of the capsule and of thin-walled capsular blood vessels. Positive immunohistochemical staining of tumour cells for thyroglobulin and negative staining for calcitonin, chromogranin and CEA allowed distinction from medullary carcinoma. Electronmicroscopy revealed groups of tumour cells, surrounded by abundant basement membrane type material. Occasional tumour cells contained abundant cytoplasmic intermediate filaments. A flow cytometric analysis revealed one tumour to have a diploid DNA pattern and the other to be DNA aneuploid. These cases illustrate that a malignant variant of hyalinizing trabecular adenoma, namely hyalinizing trabecular carcinoma, exists. Hyalinizing trabecular tumours of the thyroid should not be considered uniformly benign lesions. As with follicular neoplasms, multiple sections from the capsule should be examined histologically in order to assess the presence or absence of capsular and/or vascular invasion. PMID- 8732346 TI - Choriocarcinoma associated with transitional cell carcinoma in-situ of the ureter. PMID- 8732347 TI - Primary intracerebral leiomyoma: a case with eosinophilic inclusions of actin filaments. PMID- 8732348 TI - Localized primary amyloidosis of axillary lymph nodes. PMID- 8732349 TI - Mucinous elements in Sertoli-Leydig and granulosa cell tumours: a reevaluation. PMID- 8732350 TI - Thrombomodulin immunoreactivity in adenomatoid tumour of the uterus. PMID- 8732351 TI - Hydatidiform moles: DNA flow cytometry, image analysis and selected topics in molecular biology. PMID- 8732352 TI - Expression of PZ-peptidases by cultures of several pathogenic fungi. Purification and characterization of a collagenase from Trichophyton schoenleinii. AB - Peptidolytic activity was studied in the broken-cell extracts of 17 isolates of pathogenic fungi tested with phenylazobenzyloxycarbonyl-Pro-Leu-Gly-Pro-Arg (PZ PLGPA) as a substrate. All the fungi studied except Candida spp., Cryptococcus neoformans and two actinomycetes hydrolyzed the substrate and therefore contained a so-called PZ-peptidase activity. Of all the positive strains, Trichophyton schoenleinii, a pathogenic fungus showed the highest activity and was therefore chosen as a source for PZ-peptidase purification. The four chromatographic steps, a 'negative' dye column, a 'positive' dye column, hydroxyapatite Ultrogel, and modified TSK (HW 55), gave a highly purified peptidase with a 12% overall yield. Inhibitor studies suggested that the 82 000 M(r) PZ-peptidase is a metalloproteinase. Moreover it cleaved native rat type I collagen. Partial peptide sequencing showed a strong sequence homology to regions of two metalloproteinases previously identified in the yeast Saccharomyces cerevisiae and in rat. PMID- 8732353 TI - Dynamic expression of cell wall proteins of Candida albicans revealed by probes from cDNA clones. AB - Five cDNA clones were selected from the positive clones detected by screening a germ tube expression library constructed in lambda gt11 with rabbit antisera raised against cell wall extracts of Candida albicans. The selected clones were amplified and used to obtain affinity purified antibodies by eluting from the expressed proteins that had been previously transferred onto nitrocellulose discs. The antibodies obtained were used as probes in immunoblots of the cell wall extracts separated by denaturing polyacrylamide electrophoresis. A single protein band was detected for each clone. Detection of products of the cloned sequences varied according to the extraction procedure and/or cell morphology. These products included bands exhibiting apparent molecular weights of 40, 58, 68 and 70 kDa present in beta-mercaptoethanol (beta ME) extracts from both yeast and germ tubes, and a 30 kDa beta ME extracted protein specific for germ tubes. The expression of these products at the cell surface was confirmed by indirect immunofluorescence. Expression of the mRNAs of the different cDNA clones varied according to growth- and morphology-related factors and showed no direct correlation between expression and presence in the cell wall. These observations suggest that complex mechanisms are involved in the regulation and expression of cell surface components of C. albicans. PMID- 8732354 TI - Systemic candidiasis in Sprague-Dawley rats. AB - A reproducible model of a generalized Candida albicans infection was established in rats to allow a precise evaluation of the efficacy of antifungal compounds. In contrast to the intravenous C. albicans model in mice, which serves as a primary model for in vivo efficacy studies of antimycotic compounds, the infectious process in Sprague-Dawley rats is more severely spread into organs other than the kidneys, such as brain, heart, liver, lung, retina and spleen. Apart from a severe granulomatous nephritis beginning 1 day after infection, we observed a severe pneumonitis 3 days after infection with a mass of extravasal erythrocytes in the interstitium and the alveolar space. In addition, multiple nodular lesions could be observed in the brain, heart, liver, retina and spleen on the first day after infection. Lethality was 100% within 1 week, the majority of deaths occurring from 5 to 7 days. Antifungal therapy with amphotericin B or fluconazole led to long-term survival over 4 months, which could not be achieved in mice. PMID- 8732355 TI - Rhizomys sumatrensis and Cannomys badius, new natural animal hosts of Penicillium marneffei. AB - The incidence of Penicillium marneffei infection has increased substantially, especially in persons with HIV infection. Very little is known about the natural reservoirs or animal hosts of P. marneffei. This pathogenic fungus was first isolated from a species of bamboo rat (Rhizomys sinensis) in Vietnam and later from another rodent species, R. pruinosus. We studied a total of 75 captured bamboo rats; P. marneffei could be isolated from the internal organs of 13 of 14 (92.8%) of large bamboo rats, R. sumatrensis, and of 3 of 10 reddish-brown small bay bamboo rats, Cannomys badius (30%). All 51 greyish-black C. badius were negative on culture. Among R. sumatrensis, P. marneffei were frequently recovered from the lungs (85.7%), spleen (50%) and liver (28.6%). Of the 28 soil samples collected from the bamboo rat burrows and the 67 from the residential areas of patients with P. marneffei infection, P. marneffei was isolated from one soil sample collected from a burrow of R. sumatrensis. The mycological characteristics of P. marneffei isolates from bamboo rats and humans were very similar. Our data indicate that R. sumatrensis and C. badius may be important animal hosts of P. marneffei in northern Thailand. PMID- 8732356 TI - Molecular karyotyping of multiple yeast species isolated from nine patients with AIDS during prolonged fluconazole therapy. AB - Variations in molecular karyotype and fluconazole susceptibility of serial yeast isolates from the oral cavities of nine patients with AIDS receiving fluconazole for single or multiple episodes of oropharyngeal candidiasis were monitored. Multiple yeast species were isolated from the initial oral specimens in six patients. Molecular karyotyping identified at least eight different DNA subtypes of C. albicans, at least eight of T. glabrata and only one DNA subtype each of C. krusei, C. tropicalis and C. parapsilosis. Among isolates of T. glabrata, fluconazole MICs in each patient were consistently within one or two dilutions, regardless of strain variations. Similarly, among five patients monitored during one course of therapy, the MICs of fluconazole of C. albicans isolates of either the same or different DNA subtypes remained within two dilutions. However, increases in MICs of fluconazole of C. albicans were observed in four patients who received two or more courses of fluconazole, three of whom had the same DNA subtype and one of whom changed from one DNA subtype to another. PMID- 8732357 TI - Identification of the conserved coding sequences of three chitin synthase genes in Fonsecaea pedrosoi. AB - Primers having designs based on highly conserved stretches in the deduced amino acid sequences of chitin synthase (CHS) genes were used in PCR reactions to amplify 600 bp and 366 bp products from the genomic DNA of three major causal agents of chromoblastomycosis. Cloning and sequencing of the PCR products of one of these fungi, Fonsecaea pedrosoi, identified three CHS sequences designated as FpCHS1, FpCHS2 and FpCHS3. FpCHS1 and FpCHS2 were homologous to regions of CHS1 and CHS2 of Saccharomyces cerevisiae, and their derived amino acid sequences fell into chitin synthase classes I and II, respectively. FpCHS3 was homologous to a region of the CAL1/CSD2 gene of S. cerevisiae, which codes for the chitin synthase three (Chs3) enzyme in that fungus. Phylogenetic trees constructed using the deduced amino acid sequences of PCR-amplified CHS products from many fungi clustered F. pedrosoi with other dematiaceous fungi, providing new molecular evidence for the genetic relatedness of these organisms. The identification of these CHS genes in F. pedrosoi will facilitate future studies of the functional roles of chitin synthases in the unique in vivo dimorphism exhibited by chromoblastomycotic fungi. PMID- 8732358 TI - Natural habitat of Cryptococcus neoformans var. neoformans in decaying wood forming hollows in living trees. AB - Cryptococcus neoformans var. neoformans was repeatedly isolated from decaying wood forming hollows in living trees growing in urban areas of Rio de Janeiro, Brazil. A new natural habitat for C. neoformans var. neoformans has been found that is not associated with specific trees. PMID- 8732359 TI - Aspergillus flavus mycetoma and epidural abscess successfully treated with itraconazole. AB - Aspergillus spp. rarely cause mycetomata. We report a patient with diabetes and nephrotic syndrome with Aspergillus flavus mycetoma of the back, with the development of an epidural abscess, diskitis and vertebral osteomyelitis. The patient was successfully treated with decompressive laminectomy and a 14-month itraconazole regimen. Serial serum itraconazole levels and quantitative Aspergillus antigen levels were performed. This is the second reported and first extrapedal case of mycetoma caused by A. flavus. PMID- 8732360 TI - Genetic homogeneity of Trichophyton mentagrophytes var. interdigitale isolated from geographically distant regions. AB - To evaluate the genetic homogeneity of Trichophyton mentagrophytes var. interdigitale, restriction enzyme analysis of mitochondrial(mt) DNA was performed on 29 isolates of T. mentagrophytes var. interdigitale isolated from Belgian or Indian patients with dermatophytosis. The restriction enzyme profiles of these mtDNAs were compared with those of the teleomorphic members composing the T. mentagrophytes complex. Using the restriction enzymes MspI, HaeIII, HindIII and BglII the restriction profiles of all the examined clinical isolates showed the same profiles as those of Arthroderma vanbreuseghemii. T. mentagrophytes var. interdigitale isolates found in Japan have been shown to have the same profiles as those of A. vanbreuseghemii. Therefore, T. mentagrophytes var. interdigitale is considered to be a highly homogeneous taxon phylogenetically related to A. vanbreuseghemii. PMID- 8732361 TI - Isolation of Malassezia sympodialis from feline skin. AB - Carriage of Malassezia yeasts was investigated in 17 cats in two colonies using a lipid-supplemented culture medium. Malassezia pachydermatis was isolated from one cat. Lipid-dependent Malassezia yeasts with electrophoretic karyotypes consistent with M. sympodialis were isolated from all six cats in one group and from one of 11 in the second group. To our knowledge, this is the first report of the isolation of lipid-dependent yeasts from cats. PMID- 8732362 TI - Comparisons of the susceptibilities of planktonic and adherent Candida albicans to antifungal agents: a modified XTT tetrazolium assay using synchronised C. albicans cells. AB - Adhesion of synchronised yeast-phase Candida albicans cells to tissue culture plastic, and the susceptibility of planktonic and adherent cells to antifungal agents, was investigated using a modified tetrazolium (XTT) assay. MIC data demonstrated that ketoconazole and amphotericin B were highly active against planktonic C. albicans yeast-phase cells. XTT tetrazolium assays permitted comparisons of MIC values with XTT formazan IC50 and IC80 (percentage inhibitory concentrations); IC50 and IC80 values for amphotericin B and ketoconazole were similar. Furthermore, IC50 and IC80 values for 24 h incubation with antifungal agent were typically higher than corresponding IC50 and IC80 values for 48 h incubation. Furthermore, in comparison to values for planktonic Candida cells, adherent cells were typically less susceptible to amphotericin B and ketoconazole. For example, with increasing incubation time following the initial adhesion period, cells became progressively less susceptible to amphotericin B and ketoconazole: 24 h (P < 0.05) and 48 h (P < 0.001). Furthermore, other azoles showed the same activities compared with ketoconazole against both planktonic and adherent cells. Overall, the data demonstrate the usefulness of the XTT tetrazolium assay in describing comparisons of the susceptibility profiles for both planktonic and adherent synchronous yeast phase C. albicans in vitro. PMID- 8732363 TI - West of Scotland Coronary Prevention Study: implications for clinical practice. The WOSCOPS Study Group. PMID- 8732364 TI - QT dispersion and heart rate variability. PMID- 8732365 TI - Results of serial enzyme measurements in acute myocardial infarction support early reperfusion as the goal of therapy. PMID- 8732366 TI - What factors determine exercise capacity in left ventricular dysfunction? PMID- 8732367 TI - Alcoholic cardiomyopathy: is outcome linked to stimulus withdrawal? PMID- 8732368 TI - Thrombolysis-associated ventricular fibrillation: is it reperfusion, the drug or what? PMID- 8732369 TI - Does positron emission tomography contribute to the management of clinical cardiac problems? PMID- 8732370 TI - Interventional cardiology in pregnancy. PMID- 8732371 TI - Cellular mechanisms of contractile dysfunction in human heart failure. PMID- 8732372 TI - Fitness in the fit: does physical conditioning affect cardiovascular risk factors in middle-aged marathon runners? AB - OBJECTIVE: The study was designed to assess cardiovascular risk factors in marathon runners with different degrees of fitness. DESIGN: A total of 30 male middle-aged marathon runners were divided according to their marathon running time into fit (265 +/- 8 min), fitter (222 +/- 5 min) and fittest (178 +/-12 min). The three groups of 10 runners each were comparable in age, weight, and body surface area. Cardiovascular risk factors were assessed by measuring arterial pressure before and during exercise (150 watts) and determination of plasma lipoproteins, uric acid, glucose and white blood cell count before and after a marathon run. RESULTS: All measured laboratory values such as high density lipoprotein cholesterol (P < 0.05), low-density lipoprotein cholesterol (P < 0.05), total cholesterol (non-significant), triglycerides (non-significant), blood sugar (non-significant), uric acid (P < 0.01) and white blood cell count (P < 0.05) indicated a lower cardiovascular risk in the fastest when compared with the slowest runners. Resting blood pressure was similar in the three groups but consistently lower at all levels of exercise in the fittest when compared with the less fit runners. The fittest runners also showed greater increases in high density lipoprotein cholesterol after the marathon run (14% vs 8% in the slowest runners, P < 0.005). CONCLUSIONS: We conclude that even at the extreme end of a continuum such as represented by well-conditioned, middle-aged marathon runners, cardiovascular risk factors are related to the degree of fitness, as measured by the marathon running time. PMID- 8732373 TI - Determinants of exercise capacity in patients with coronary artery disease and mild to moderate systolic dysfunction. Role of heart rate and diastolic filling abnormalities. AB - BACKGROUND: To test the hypothesis that diastolic filling abnormalities are an important cause of exercise limitation in some patients with coronary artery disease we assessed the factors limiting exercise capacity in a group of patients with coronary artery disease in whom exercise limitation was greater than expected from the degree of resting left ventricular systolic dysfunction. METHODS AND RESULTS: We assessed the relationship between exercise capacity (maximal oxygen consumption) during erect cycle ergometry, heart rate, radionuclide indices of left ventricular systolic function (ejection fraction) and diastolic filling (peak filling rate, and time to peak filling) during semi erect cycle ergometry in 20 patients (15 male, five female) who were aged 42-72 years (mean 61 years) and had angiographically proven coronary artery disease and evidence of reversible myocardial ischaemia on thallium scintigraphy. All patients exhibited marked exercise limitation (maximal oxygen consumption 8.7- 22.4 ml.min-1.kg-1--mean 15.9 ml.kg-1.min-1, which was 61.1 +/- 16% of age and gender predicted maximum) due to breathlessness or fatigue rather than angina, in spite of a mean ejection fraction for the group of 46.5% (range 30-67%). We also compared the diastolic filling characteristics of these patients during exercise with 10 healthy controls (age 38-66, mean 58 years; eight male, two female). Comparing diastolic filling characteristics, peak filling rate was higher and time to peak filling shorter both at rest and at peak exercise in controls than patients (peak filling rate 3.1 +/- 0.5 vs 2.2 +/- 0.9 EDV.s-1, P = 0.01 at rest and 8.3 +/- 0.8 vs 5.2 +/- 1.9 EDV.s-1, P < 0.0001 on exercise; time to peak filling 115.2 +/- 29.8 vs 228.9 +/- 71.7 ms, P < 0.0001 at rest and 52.8 +/- 16.2 vs 139.6 +/- 44.8 ms, P < 0.0001 on exercise respectively). On univariate analysis in the patients studied, maximal oxygen consumption was correlated with peak heart rate (r = 0.45 P = 0.04), peak exercise time to peak filling (r = 0.85 P < 0.0001), peak exercise peak filling time rate (r = 0.51 P = 0.019), and the relative increase in cardiac output i.e. cardiac output peak/cardiac output rest (r = 0.58, P = 0.008). There was no correlation between maximal oxygen consumption and resting indices of diastolic filling (peak filling rate and time to peak filling) or with resting or peak exercise ejection fraction. On multiple regression analysis, only peak exercise time to peak filling was significantly related to maximal oxygen consumption. CONCLUSION: We have observed a strong correlation between exercise capacity and indices of exercise left ventricular diastolic filling, and have confirmed previous studies showing a poor correlation with resting and exercise indices of systolic function and resting diastolic filling, in patients with coronary artery disease. PMID- 8732374 TI - Ventricular fibrillation following administration of thrombolytic treatment. The EMIP experience. European Myocardial Infarction Project. AB - Reperfusion-induced ventricular fibrillation has been demonstrated in animal models of myocardial ischaemia, but no evidence exists for this in humans. The European Myocardial Infarction Project compared the efficacy and safety of pre hospital thrombolytic therapy with that of hospital therapy. The objective of this study was to investigate the occurrence of reperfusion-induced ventricular fibrillation in acute myocardial infarction patients following thrombolytic therapy. In a double-blind multicentre trial, eligible patients were randomized to receive anistreplase at home followed by placebo in the hospital (A/P group), or placebo followed by anistreplase (P/A group). The occurrence of ventricular fibrillation, and other adverse events were recorded on specific study forms and could be attributed to defined time intervals. The incidence of ventricular fibrillation in the A/P group was significantly higher following the pre-hospital injection than in the P/A group (2.5% vs 1.6%; P = 0.021); the situation was reversed following the hospital injection (3.6% vs 5.3%; P = 0.002). No relationship was found between this excess of ventricular fibrillation and the patients' condition, with the exception of the site of the infarct. These results suggest the existence of reperfusion-induced ventricular fibrillation in patients developing myocardial infarction who receive thrombolytic treatment. PMID- 8732375 TI - Dobutamine stress echocardiography after coronary artery bypass grafting. Transthoracic vs biplane transoesophageal imaging. AB - Graft failure or progressive native vessel disease can be a serious problem after coronary artery bypass grafting. However, because of poor image quality it may be difficult to evaluate these patients by transthoracic stress echocardiography. The purpose of this study, therefore, was to evaluate the effectiveness of dobutamine stress echocardiography in the detection of myocardial territories with compromised vascular supply (due to either an obstructed native vessel without graft, and obstructed graft, or a native vessel obstructed distal to bypass graft insertion with < or = 50% luminal diameter reduction on angiography) after coronary artery bypass grafting and to determine additional information obtained by biplane transoesophageal stress echocardiography. Sixty patients (54 men, mean age 59 +/- 8.5 years) who had undergone coronary bypass grafting (total number of graft vessels 198) were evaluated from 6 months to 14 years (mean 6.2 years) after surgery. Transthoracic dobutamine stress echocardiography, biplane transoesophageal dobutamine stress echo, and coronary angiography were performed and evaluated by independent examiners. An infusion of dobutamine up to a maximum of 40 micrograms.kg-1.min-1 was administered, and additional atropine (0.25-1.0 mg) was given if 85% of age-predicted maximal heart rate was not reached. Biplane transoesophageal echocardiography was performed in the transgastric short-axis view as well as transoesophageal 4- and 2-chamber views, allowing division of the left ventricle into a 14-segment scheme. Wall motion abnormalities induced with dobutamine stress were used to predict regional vascular insufficiency. A 4-point scale, ranging from 'excellent' (1) to 'impossible' (4) was used to assess each system's ability to evaluate all left ventricular segments. Forty-five patients, of whom 35 were identified by transthoracic echocardiography (sensitivity 78%), had at least one territory with a compromised vascular supply. In 15 patients, the vascular supply was uncompromised, with 13 showing no wall motion abnormalities inducible by transthoracic echocardiography (specificity 86%). However, biplane transoesophageal echocardiography had a higher sensitivity and specificity than transthoracic echocardiography in detecting compromised vascular supply, 93% and 93%, respectively. The former system correctly classified the vascular supplies in 113 of 120 vascular territories (94%), according to whether they were compromised or uncompromised. This was significantly more (P < 0.05) than by classification with transthoracic dobutamine echocardiography, by which system only 102 of the 120 vascular territories were correctly assessed (85%). Compared with the conventional transgastric monoplane short-axis view, examination using three different views via a biplane probe results in a higher sensitivity (93% vs 84%). Assessed on a 4-point scale, the ability to evaluate all left ventricular segments was 2.3 +/- 0.7 (mean +/- SD) for transthoracic echocardiography and 1.7 +/- 0.7 (P < 0.01) for biplane transoesophageal echocardiography. After coronary artery bypass grafting transthoracic dobutamine stress echocardiography has acceptable accuracy in the detection of regional vascular insufficiency. However, this accuracy can be improved using the higher image quality of transoesophageal echocardiography, combined with the advantages of several different views obtained by biplane transoesophageal echocardiography. PMID- 8732376 TI - Late activation of the fibrinolytic system in myocardial infarction treated with thrombolytic therapy. Influence of the coronary anatomical substrate. AB - Procoagulant activity, thrombin and fibrinolytic system activation have been demonstrated in the first 24-48 h after acute myocardial infarction treated with thrombolytic therapy. Little is known about what happens in the subsequent days, during which the incidence of ischaemic recurrence is high. In 21 patients treated with streptokinase and in 20 patients treated with urokinase we evaluated, with multiple plasma determinations, D-dimer and fibrinogen plasma levels in the first week after myocardial infarction. From the 2nd hour after the beginning of thrombolysis to the 4th day, all patients received intravenous heparin in doses sufficient to raise the partial thromboplastin time to twice its normal level; subcutaneous calcium heparin (12,000 U/day) was subsequently substituted for the intravenous route. Coronary angiography was performed 7 days after infarction. From the basal values 2.22 +/- 1.44 nmol.1(-1) in the streptokinase group and 3.28 +/- 3.05 nmol.1(-1) in the urokinase group, D-dimer rose consistently in the 1st hour after thrombolysis 269.4 +/- 206.7 nmol.1(-1) and 44.5 +/- 35.5 nmol.1(-1) in the streptokinase and urokinase groups, respectively; P < 0.001. After the peak value, which in both groups was reached after 5 h, D-dimer slowly decreased during the study period. It reverted to normal values only in 10/21 patients in the streptokinase group; in the urokinase group normalization was attained in 14/20 patients between the 3rd and 6th days. After withdrawal of i.v. heparin in patients of both groups with TIMI 0 or 1 grade of coronary patency, D-dimer rose to levels four to seven times greater than normal; in patients of both groups with TIMI 2 or 3 grade coronary flow, D dimer showed a monophasic pattern of progressive normalization (P < 0.05 and P < 0.01 at the 6th and 7th days, respectively, for differences between TIMI 0-1 and TIMI 2-3 groups). After myocardial infarction, thrombolysis is followed by active and persistent fibrin degradation more marked and lasting after streptokinase than after urokinase. When occurring sooner, it is a consequence of plasmin activation induced by thrombolytic agents; later it seems to be related to intracoronary substrate, as suggested by the relationship of plasma elevation of D-dimer with the presence of occluded or suboccluded infarction-related vessels. PMID- 8732377 TI - Differential effects of tissue plasminogen activator and streptokinase on infarct size and on rate of enzyme release: influence of early infarct related artery patency. The GUSTO Enzyme Substudy. AB - BACKGROUND: The recent international GUSTO trial of 41,021 patients with acute myocardial infarction demonstrated improved 90-min infarct related artery patency as well as reduced mortality in patients treated with an accelerated regimen of tissue plasminogen activator, compared to patients treated with streptokinase. A regimen combining tissue plasminogen activator and streptokinase yielded intermediate results. The present study investigated the effects of treatment on infarct size and enzyme release kinetics in a subgroup of these patients. METHODS: A total of 553 patients from 15 hospitals were enrolled in the study. Four thrombolytic strategies were compared: streptokinase with subcutaneous heparin, streptokinase with intravenous (i.v.) heparin, tissue plasminogen activator with i.v. heparin, and streptokinase plus tissue plasminogen activator with i.v. heparin. The activity of alpha-hydroxybutyrate dehydrogenase (HBDH) in plasma was centrally analysed and infarct size was defined as cumulative HBDH release per litre of plasma within 72 h of the first symptoms (Q(72)). Patency of the infarct-related vessel was determined by angiography in 159 patients, 90 min after treatment. RESULTS: Infarct size was 3.72 g-eq.1(-1) in patients with adequate coronary perfusion (TIMI-3) at the 90 min angiogram and larger in patients with TIMI-2 (4.35 g-eq.1(-1) or TIMI 0-1 (5.07 g-eq.1(-1) flow (P = 0.024). In this subset of the GUSTO angiographic study, early coronary patency rates (TIMI 2 + 3) were similar in the two streptokinase groups (53 and 46%). Higher, but similar, patency rates were observed in the tissue plasminogen activator and combination therapy groups (87 and 90%). Median infarct size for the four treatment groups, expressed in gram-equivalents (g-eq) of myocardium, was 4.4, 4.5, 3.9 and 3.9 g-eq per litre of plasma (P = 0.04 for streptokinase vs tissue plasminogen activator). Six hours after the first symptoms, respectively 5.3, 6.6, 14.0 and 13.6% of total HBDH release was complete (P < 0.0001 for streptokinase vs tissue plasminogen activator). CONCLUSIONS: Rapid and complete coronary reperfusion salvages myocardial tissue, resulting in limitation of infarct size and accelerated release of proteins from the myocardium. Treatment with tissue plasminogen activator, resulting in earlier reperfusion was more effective in reducing infarct size than the streptokinase regimens, which contributes to the differences in survival between treatment groups in the GUSTO trial. PMID- 8732378 TI - Isolated acute occlusion of a large right ventricular branch of the right coronary artery following coronary balloon angioplasty. The only true 'model' to study ECG changes in acute, isolated right ventricular infarction. AB - An isolated right ventricular infarction occurs rarely and data on its electrocardiographic appearance and underlying angiographically proven cause are scarce. The electrocardiographic response of acute right ventricular ischaemia is often obscured by the coexisting forces of the ischaemic mass of the inferior wall of the left ventricle when the right coronary artery itself becomes occluded. Percutaneous transluminal coronary angioplasty of the right coronary artery may cause an isolated occlusion of a right ventricular branch. We encountered this phenomenon in nine patients. In all, it led to acute isolated right ventricular ischaemia with ST elevations in the right precordial leads (V1 V3, V3R and V4R) on the electrocardiogram. We conclude that the ECG pattern of pure right ventricular ischaemia can be seen when an isolated occlusion of a large right ventricular branch occurs, for example as a complication of percutaneous transluminal coronary angioplasty. PMID- 8732379 TI - Differences of disease progression in congestive heart failure due to alcoholic as compared to idiopathic dilated cardiomyopathy. AB - In patients with alcoholic cardiomyopathy there is evidence that mild heart failure is reversible if patients abstain from alcohol, but there is no consensus whether the disease is progressive once structural myocardial dilation has evolved. The aim of the present study was to compare the long-term course of congestive heart failure due to alcoholic and idiopathic dilated cardiomyopathy. Of 75 patients with overt congestive heart failure, 23 had alcoholic cardiomyopathy and were compared to 52 patients with idiopathic cardiomyopathy. The mean age was 48 +/- 12 years. Despite medical therapy, heart failure class New York Heart Association III-IV was present in 52% of patients with alcoholic and 47% of patients with idiopathic cardiomyopathy (not significant). Their mean left ventricular ejection fraction was 30 +/- 12% vs 28 +/- 12% and left ventricular end-diastolic volumes were 264 +/- 125 ml and 254 +/- 100 ml respectively (not significant). Overall survival at 1, 5 and 10 years was 100%, 81% and 81% for the group with alcoholic dilated cardiomyopathy and 89%, 48% and 30% for the group with idiopathic cardiomyopathy, respectively (P = 0.041), and the difference was even greater for transplant-free survival P = 0.005). Clinical and invasive signs of left and right heart failure as well as left ventricular dimensions were predictive of a fatal outcome; however, symptom duration and left ventricular volumes were only predictive in patients with idiopathic cardiomyopathy, suggesting that in the two patient groups different mechanisms may lead to death. Mortality in patients with severe congestive heart failure and left ventricular dilatation due to alcoholic cardiomyopathy is significantly lower than that in patients with idiopathic cardiomyopathy and similar degrees of heart failure. Thus, despite structural changes inherent in marked left ventricular dilatation, disease progression in alcoholic dilated cardiomyopathy is different from that in idiopathic cardiomyopathy and thus may have implications for the choice of therapy. PMID- 8732380 TI - QT dispersion and RR variations on 12-lead ECGs in patients with congestive heart failure secondary to idiopathic dilated cardiomyopathy. AB - Increased QT dispersion, which has been proposed as a marker of ventricular repolarization inhomogeneity, may predispose to ventricular arrhythmias. Data on QT dispersion in patients with congestive heart failure are scarce. In this study, conventional 12-lead ECGs were recorded in 135 consecutive patients with congestive heart failure secondary to idiopathic dilated cardiomyopathy. Seventy five patients were excluded from QT interval assessments due to one or more of the following reasons: (1) low amplitude of the T wave (n = 3), (2) atrial fibrillation (n = 26) and (3) bundle branch block (n = 46). QT dispersion was calculated as (1) QT-range: the difference between the maximum and minimum QT intervals on any of the 12 leads and (2) QT-SD: the standard deviation of the QT interval in all the 12 leads. RR intervals were measured in leads II, aVL, V2 and V5. QT-SD (20.85 +/- 5.00 ms) was significantly (r = 0.8997, P < 0.001) related to QT-range (65.65 +/- 15.77 ms), but not to the QT interval. Neither QT-range nor QT-SD was significantly related to age, left ventricular dimensions, left ventricular end diastolic pressure, left ventricular ejection fraction or left ventricular wall thickness. There was no significant difference in QT dispersion between survivors and those who died (n = 8) or were transplanted (n = 9) during 34 +/- 23 month follow-up. No significant difference in QT dispersion was observed between patients with and without ventricular tachycardia (> or = three consecutive beats) detected on 24-h Holter ECGs. RR interval variation was significantly lower in patients who died compared with survivors (standard deviation: 10.37 +/- 3.61 vs 36.02 +/- 35.03 ms, P < 0.001; coefficient of variance: 1.87 +/- 0.7% vs 4.50 +/- 4.9%, P = 0.001). This was also true in patients with bundle branch block. These observations suggest that QT dispersion in idiopathic dilated cardiomyopathy is not significantly related to either QT interval or cardiac size and function and does not predict death. The application of QT dispersion assessment is limited by the commonly encountered atrial fibrillation and bundle branch block in this patient population. However, reduced RR variation on standard 12-lead ECGs has important prognostic implications in these patients. PMID- 8732381 TI - Prognostic value of late potentials in patients with congestive heart failure. AB - To investigate whether detection of ventricular late potentials could provide prognostic information in patients with congestive heart failure with or without bundle branch block, we prospectively obtained a signal-averaged ECG from 151 patients with congestive heart failure, using specific criteria in 57 patients with bundle branch block. Late potentials were detected in 49 patients (32.5%); their incidence was not significantly different in patients without (31%; 29 patients) or with bundle branch block (35%; 20 patients). Late potentials were present in 25 of 73 patients (34%) with idiopathic dilated cardiomyopathy, in 20 of 57 patients (35%) with ischaemic cardiomyopathy and in four of 21 patients (19%) with hypertensive heart disease (ns). Age, NYHA class, ejection fraction and use of amiodarone were not statistically different among patients with or without late potentials. In contrast, patients with late potentials had more past episodes of sustained ventricular tachycardia (8.2%; four patients) than those without late potentials (1.9%; two patients). Twenty four hour ambulatory ECGs were obtained in 135 patients (89%). Non-sustained ventricular tachycardia was not correlated with the presence of late potentials found in 45 of 88 patients (51%) without late potentials and in 29 of 47 patients (62%) with late potentials (ns). The mean follow-up was 27 +/- 12 months; 51 patients died, 31 from progressive congestive heart failure and 13 suddenly; seven prospectively had sustained ventricular tachycardia. The total mortality rate, the cardiac mortality rate and sudden death risk were not significantly related to the presence of late potentials; their incidence were respectively 35% (36 patients), 32% (33 patients) and 10% (10 patients) in patients without late potentials and 31% (15 patients), 23% (11 patients) and 6% (three patients) in those without late potentials. The incidence of sustained ventricular tachycardia during follow up was 2% (two patients) in patients without late potentials and 10% (five patients) in those with late potentials. The incidence of sustained ventricular tachycardia experienced by the patients before the study or seen during follow-up was significantly increased in the presence of late potentials: 18% (nine patients) vs 2% (two patients) in the absence of late potentials (P < 0.001). Removal from the study of data from patients with bundle branch block, patients with severe congestive heart failure (NYHA 3 or 4) or patients taking amiodarone did not alter these results. Thus, signal-averaged ECG results only improved risk stratification for sustained ventricular tachycardia in patients with congestive heart failure and failed to identify patients at high risk for sudden death. PMID- 8732382 TI - Comparative value of Doppler echocardiography and cardiac catheterization for management decision-making in patients with left-sided valvular regurgitation. AB - OBJECTIVE: The purpose of this study was to examine the value of non-invasive clinical and Doppler echocardiographic findings, compared to cardiac catheterization, in management decision-making for patients with left-sided valvular regurgitation. METHODS: One hundred and thirty-five consecutive patients with left-sided valvular regurgitation who underwent cardiac catheterization and detailed Doppler echocardiography were prospectively studied. Two independent groups of experienced cardiologists, given clinical information combined with either Doppler echocardiographic or cardiac catheterization data, decided to operate, not to operate, or remained uncertain. RESULTS: In 63 (81%) of 78 patients with mitral regurgitation, there was agreement on the decision for valve surgery or medical treatment between Doppler echocardiography and cardiac catheterization. Valve repair was performed in 22 patients, which agreed with the echocardiographic decision. In the remaining 15 patients, although the severity and type of mitral valve lesions and left ventricular functional status were confirmed by Doppler echocardiography, the clinical decision was uncertain; additional information concerning coronary anatomy (13 patients) and pulmonary artery pressure (one patient) or both (one patient) was required. In 47 of 57 patients (82%) with aortic regurgitation, there was agreement on their management as a result of Doppler echocardiography and cardiac catheterization findings. In 10 patients, the clinical decision reached with the help of Doppler echocardiography alone was uncertain and coronary (seven patients), left ventricular (two patients) angiography or aortography (one patient) were requested. Overall, there were no conflicting clinical decisions made by the two methods in patients with either mitral or aortic regurgitation. CONCLUSIONS: In every patient in whom it was considered that a decision could be reached by echocardiography alone (more than 80% of patients) there was 100% agreement from the cardiac catheterization assessment group on the management decision. Therefore, in patients with significant mitral or aortic regurgitation where echocardiographic data is adequate, cardiac catheterization can be safely omitted from the investigative process for surgery. Where echocardiographic indices are conflicting, or significant coronary artery disease is suspected, cardiac catheterization is required. PMID- 8732383 TI - Clinical outcome and echocardiographic findings of native and prosthetic valve endocarditis in the 1990's. AB - Prosthetic valve endocarditis is considered to be associated with a more severe prognosis than native valve endocarditis. Among other factors, inappropriate visualization of vegetations in prosthetic valve endocarditis by transthoracic echocardiography is responsible for this observation. Since the introduction of transoesophageal echocardiography into clinical practice the diagnostic sensitivity and specificity of the detection of vegetations located on prosthetic valves have been enhanced. Therefore we aimed to determine and compare the prognosis of prosthetic valve endocarditis and native valve endocarditis in the era of this improved diagnostic approach. One hundred and six episodes of infective endocarditis in 104 patients were seen at our institution between 1989 and 1993. Eighty patients (77%) had native valve endocarditis and 24 (23%) had late prosthetic valve endocarditis. In the latter group two patients had recurrent infective endocarditis. Patients with prosthetic valve endocarditis were older (mean age 64 vs 54 years in native valve endocarditis; P < 0.001) and the majority was female (62% vs 38% in native valve endocarditis; P < 0.05). In prosthetic valve endocarditis, infection of a valve in the mitral position predominated (65% vs 30% in native valve endocarditis; P < 0.01), whereas in native valve endocarditis more than half the cases had isolated aortic valve endocarditis (51% vs 27% in prosthetic valve endocarditis; P < 0.01). In prosthetic valve endocarditis more cases were caused by Staphylococcus aureus (31% vs 14% in native valve endocarditis; P = 0.08), whereas in native valve endocarditis the most frequent organisms were streptococci (29% vs 19% in prosthetic valve endocarditis; P = 0.12). Differences in the clinical features of native valve endocarditis and prosthetic valve endocarditis could not be found except for a higher rate of embolism in native valve endocarditis (40% vs 19% in prosthetic valve endocarditis; P < 0.05). Vegetations could be detected by transthoracic echocardiography more frequently in native valve endocarditis (71% vs 15% in prosthetic valve endocarditis; P < 0.0001). Transoesophageal echocardiography visualized vegetations in 95% of the episodes of native valve endocarditis and in 80% of the episodes of prosthetic valve endocarditis (P = 0.09). Thus, the diagnostic gain by transoesophageal echocardiography was greatest in prosthetic valve endocarditis. Patients with native valve endocarditis had significantly larger vegetations than patients with prosthetic valve endocarditis (P < 0.05 for length, P < 0.001 for width). The median time to diagnosis was similar in native valve endocarditis and prosthetic valve endocarditis (31 vs 28 days). Surgery was performed in 74% of patients with native valve endocarditis and in 58% of those with prosthetic valve endocarditis; the median time delay between the diagnosis of infective endocarditis and surgery tended to be shorter in prosthetic valve endocarditis than in native valve endocarditis (45 vs 60 days). The in-hospital mortality and the mortality during a follow-up of 22 +/- 10 months did not significantly differ between native valve endocarditis and prosthetic valve endocarditis (21% vs 17%; 28% vs 25%). In summary in the era of transoesophageal echocardiography, late prosthetic valve endocarditis does not seem to carry a worse prognosis than native valve endocarditis. This can be attributed in part to the improved diagnostic accuracy achieved by transoesophageal echocardiography leading to comparable diagnostic latency periods in both patient groups. Finally, better characterization of vegetations on prosthetic valves by transoesophageal echocardiography allows early lifesaving surgery in patients with prosthetic valve endocarditis. PMID- 8732384 TI - Diagnostic accuracy of dobutamine stress echocardiography for detection of coronary heart disease in hypertensive patients. AB - To compare the diagnostic accuracy between dobutamine echocardiography and treadmill exercise electrocardiography in detecting coronary artery disease in hypertensive patients, 43 patients without electrocardiographic evidence of left ventricular hypertrophy and basal ST-T changes, who had also undergone coronary angiography, were further evaluated by dobutamine echocardiography. The patients also underwent treadmill exercise echocardiography. Left ventricular mass index was calculated by echocardiography. Twenty-nine patients had coronary artery disease, of whom 22 had multi-vessel disease and 14 a normal coronary anatomy. Twenty-eight patients had an increased left ventricular mass index. The sensitivities of dobutamine echocardiography and exercise electrocardiography for detecting coronary artery disease were 93% and 72% (P = 0.08), respectively, and the specificities were 100% and 29% (P < 0.005), respectively. Logistic regression analysis showed exercise electrocardiography to be a poor predictor of coronary artery disease (P < 0.09) but dobutamine echocardiography was significantly better (P < 0.001). When patients with increased left ventricular mass index were excluded, prediction of coronary anatomy by exercise electrocardiography improved only marginally (p = 0.4) while dobutamine echocardiography was significantly better (P < 0.001). Thus dobutamine echocardiography is superior to exercise electrocardiography for diagnosis of coronary artery disease in hypertensive patients. PMID- 8732385 TI - Residual myocardial perfusion in reversibly damaged myocardium by dipyridamole contrast echocardiography. AB - In patients with previous myocardial infarction and left ventricular asynergy, dipyridamole infusion may have the capacity to unmask myocardial viability through transient recovery of contractile function in asynergic segments. The purpose of this study was to assess simultaneous changes in myocardial perfusion and LV function--elicited by dipyridamole infusion--in infarcted, asynergic segments. The echo contrast agent Albunex was injected into the left coronary artery of 19 patients (17 males, age 49-70 years) with previous myocardial infarction and baseline left ventricular asynergy, both before and after dipyridamole infusion (up to 0.56 mg.kg-1, i.v.). Analysis was not possible in three patients due to inadequate image quality and in two due to weak contrast. There were no major adverse events, or changes in vital signs or demonstrated on the electrocardiogram. After dipyridamole, 7/14 patients, showed an improvement in regional function of asynergic segments ('responders'), whereas seven patients did not ('non-responders'). Among non-responders, five had a myocardial perfusion deficit corresponding to 41% of the total left ventricular area before dipyridamole and to 38% after dipyridamole. No baseline perfusion deficits were observed in the remaining two non-responders; one of these, however, developed transient asynergy and perfusion deficit after dipyridamole. Among responders, five showed a normal perfusion pattern, both before and after dipyridamole, while the remaining two showed a perfusion deficit which markedly decreased after dipyridamole (from 32% to 13% of total left ventricular area). Thus, residual contractile reserve of asynergic, infarcted ventricular segments appears to be associated with myocardial perfusion either preserved at baseline or recruitable by a coronary dilator stimulus. PMID- 8732386 TI - The influence of Albunex on the pulmonary circulation in patients with pulmonary hypertension or left heart failure. AB - To determine the safety of the ultrasound contrast agent Albunex, its influence on right and left heart haemodynamics in patients with pulmonary artery hypertension or left heart failure was assessed after intravenous injection. Patients with a left ventricular ejection fraction smaller than 40% or a systolic pulmonary artery pressure greater than 40 mmHg received 0.08 and 0.22 ml.kg-1 Albunex and 10 ml albumin in random order during right heart catheterization and transthoracic echocardiography. Right atrial, systolic and diastolic pulmonary artery and capillary wedge pressures were measured at 3 min and 5 min and cardiac output at 5 min after the intravenous injection of Albunex and control. The mean differences of pre- and postinjection values and their confidence intervals were tabulated and significance was anticipated if the confidence interval did not include 0. Significant changes to pre-injection values could be observed in diastolic pulmonary artery pressure 5 min after the injection of albumin and 0.08 ml.kg-1 Albunex, and in right atrial pressure 5 min after the injection of 0.22 ml.kg-1 Albunex only. Since intermediate opacification of the left ventricle was seen in only four patients with 0.22 ml.kg-1 Albunex, in the patients studied higher doses of Albunex and their safety need to be assessed. PMID- 8732387 TI - Endothelium-dependent relaxation of rat thoracic aorta by amrinone-induced nitric oxide release. AB - To determine whether amrinone and its induced increase of cyclic AMP releases nitric oxide and enhances endothelium-dependent vascular relaxation, we studied nitric oxide production and vascular relaxation of rat thoracic aorta on treatment with amrinone and forskolin, an activator of adenylate cyclase. When 20 microM amrinone was applied to ring segments of aorta previously contracted with phenylephrine, relaxation was greater in segments with endothelium than in those without (% relaxation 94 +/- 4 vs 37 +/- 7%, P < 0.01). However, a higher concentration of amrinone (> 50 microM) induced the same degree of relaxation in ring segments with or without endothelium, probably due to its direct effect on vascular smooth muscle. The maximal relaxant concentration (100 microM) of amrinone induced an increase (2.5 fold) in cyclic GMP in ring segments. Forskolin also induced concentration-dependent relaxation, but removal of the endothelium attenuated the relaxation induced by forskolin about seven-fold. NG-nitro L arginine reversed the relaxation induced by amrinone or forskolin in ring segments with, but not without, endothelium. Nitric oxide production in ring segments of aorta, following application of amrinone or forskolin, was detected by nitric oxide-selective electrode and electron paramagnetic resonance spin trapping methods. Pre-treatment with NG-nitro L-arginine or removal of the endothelium suppressed nitric oxide production by amrinone or forskolin. These data showed that amrinone enhances the release of nitric oxide from rat aortic endothelial cells, and induces endothelium-derived relaxing factor/nitric oxide mediated vasodilation. PMID- 8732388 TI - Myocardial infarction with normal coronary arteries in S/beta thalassaemia patients. PMID- 8732389 TI - Comprehensive analysis of aortic valve vegetation with anyplane, paraplane, and three-dimensional echocardiography. PMID- 8732390 TI - Successful thrombolysis treatment of a spontaneous left main coronary artery dissection without subsequent surgery. PMID- 8732391 TI - The role of neurophysiological investigation in traumatic brachial plexus lesions in adults and children. PMID- 8732393 TI - Lengthening of the forearm by callus distraction. AB - Ten patients aged 3 to 13 years (mean, 9 years and 7 months) underwent forearm lengthening by callotasis. The indications for lengthening were shortening and/or deformity of the forearm due to exostosis of the distal ulna in five cases, enchondroma of the distal ulna in one, growth disturbance after fracture of the distal radius in one, radial club hand in one, congenital amputation of the forearm in one and congenital dislocation of the radial head in one. Four had lengthening of the ulna, one of the radius and five of both the radius and the ulna. The average lengthening achieved was 30 mm. Complications encountered were pin track discharge in three cases, callus fracture in five, delayed consolidation of the callus in one and no callus formation in one. Review after 1 to 7 years follow-up (with a mean of 4 years and 9 months) showed satisfactory improvement in appearance and function especially in patients who had tumorous conditions or traumatic epiphyseal arrest. PMID- 8732392 TI - Opportunistic mycotic infections of the upper limb. A review. AB - Opportunistic fungal infections of the upper limb are rare and require a host who is immunologically compromised. A review of these rare mycotic infections is presented. The hand surgeon should be aware of these infections and should have a high index of suspicion in order to establish an early diagnosis and institute an appropriate management. PMID- 8732394 TI - Constrictive paralysis of the posterior interosseous nerve without external compression. AB - Four cases of constrictive neuropathy of the posterior interosseous nerve (PIN) in the absence of external compression are reported. All patients presented with a history of severe elbow pain with no apparent cause, followed by paralysis of the PIN. There were one or two well-localized constrictions on the PIN proximal to the arcade of Frohse where no obvious external compressive structure was observed. After epineurotomy with or without neurorrhaphy, three patients had a complete return of motor function within 1 year. The remaining patient required tendon transfer after resection of the abnormal segment of nerve. PMID- 8732395 TI - Ulnar lengthening and radial recession procedures for Kienbock's disease. Long term clinical and radiographic follow-up. AB - Twenty patients with Kienbock's disease who had undergone a joint levelling procedure, were reviewed at a mean of 11 years following surgery. Clinical and radiological evaluation revealed good long-term results with all patients complaining of less pain than before surgery and with statistically significant increases in range of motion and grip strength. Radiologically a significant number of patients were found still to have lunate fractures and fragmentation. In addition, 13 patients have developed new bone formation in the lunate fossa, and 12 show osteoarthritic changes, particularly at the radioscaphoid interface. Radial shortening was felt to be superior to ulnar lengthening in that there were fewer problems with bone union. PMID- 8732396 TI - Radiographic assessment of the trapezial space before and after ligament reconstruction and tendon interposition arthroplasty. AB - A radiographic method was developed, the trapezial space ratio, for assessing the space occupied by the trapezium (a space defined by the distal scaphoid and thumb metacarpal base divided by the thumb proximal phalanx). This method was applied to 100 normal thumb radiographs and to the radiographs of 15 patients with symptomatic degenerative arthrosis of the thumb basal joint before and after operative treatment with ligamentous reconstruction and tendon interposition arthroplasty. The trapezial space ratio averaged 0.476 +/- 0.033 for radiographs of normal thumbs, 0.372 +/- 0.084 for the pre-operative radiographs of thumbs with symptomatic basal joint arthrosis, and 0.270 +/- 0.078 for the radiographs of thumbs following basal joint arthroplasty. A significant reduction in the trapezial space ratio was noted when values from arthritic thumbs were compared to those of normal thumbs (22%; P < 0.0001). A further reduction in the trapezial space ratio was noted when post-operative values were compared to pre-operative ones (27%; P < 0.0002). Comparing post-operative trapezial space ratio values to values obtained in normal thumbs, a reduction of 43% was found in those thumbs treated operatively. These finding indicate that the trapezial space is reduced significantly in thumbs with severe degenerative arthrosis compared to normal thumbs and that ligament construction tendon interposition arthroplasty is not entirely successful in either restoring or maintaining the length of the thumb ray. PMID- 8732397 TI - First carpometacarpal joint arthritis. A comparison of two arthroplasty techniques. AB - Trapeziectomy followed by reconstruction of the intermetacarpal ligament using a small slip of extensor carpi radialis longus tendon and a wide slip of flexor carpi radialis for tendon interposition (group A) is compared with trapeziectomy followed by palmaris longus tendon interposition (group B). Group A included 11 hands in nine patients. Group B included eight hands in eight patients. Significant pain at the base of the thumb and 1st carpometacarpal joint degeneration were indications for surgery. Outcome was assessed by subjective questioning, objective clinical measurement and radiography. Group A appeared to have a slower recovery but later developed a stronger hand, better dynamic thumb function despite reduced movement and the resting trapezial space was better preserved. Calcification within the trapezial space correlated with better results but this took 3 years to occur. Patients in group B had greater relief of pain and were more consistently satisfied at an early stage. PMID- 8732398 TI - Excision of the distal ulna in rheumatoid arthritis. Is the price too high? AB - Patients with rheumatoid arthritis who underwent excision of the distal ulna were reviewed and the operated wrist was compared with the non-operated side in the 40 patients who had the procedure performed unilaterally. Radiological assessment showed that the radiocapitate measurement of carpal translocation was the most consistent and that excision of the distal ulna was not associated with statistically significant collapse, ulnar translocation or radial rotation of the carpus. 61% of wrists spontaneously developed a radial shelf or limited radiocarpal fusion following excision of the distal ulna, compared to 21% of non operated wrists. However, there was no statistically significant difference in carpal collapse or ulnar translocation between these two groups. PMID- 8732399 TI - Midcarpal instability: is capitolunate instability pattern a clinical condition? AB - Five cases are presented with clinical findings of capito-lunate instability pattern of the wrist. All painful areas and tender points were dorsal, but variable in location and intensity. All plain radiographs and fluoroscopic instability series were normal. None of the cases had an explanation for the dorsal wrist pain other than a positive dorsal capitate-displacement test. Four out of five cases were treated in a cast for 4 weeks and two had subsequent splint immobilization. Although at short-term follow-up two of these five patients became pain-free, none was completely pain-free at long-term follow-up. Three patients treated with a cast had long-term follow-up. Only one could perform his original work. These findings support a clinical condition of midcarpal instability producing dorsal wrist pain reproduced with a simple stress test. Conservative, non-operative treatment will not usually produce long-term pain relief. PMID- 8732400 TI - A prospective, randomized study with an independent observer comparing open carpal tunnel release with endoscopic carpal tunnel release. AB - In order to define the role of two-portal endoscopic carpal tunnel release, a prospective randomised study with an independent observer was performed to compare endoscopic and open surgery. Thirty-two hands in 29 patients, with symptoms, clinical signs and EMG changes consistent with idiopathic carpal tunnel syndrome were randomised to either endoscopic carpal tunnel release or open release. No significant difference in sick leave between the two groups could be found, being a mean of 17 days (range 0-31 days) with endoscopic surgery, and 19 days (range 0-42 days) with open conventional surgery. No differences in surgical results were found, but three patients in the endoscopic group suffered transient numbness on the radial side of the ring finger. PMID- 8732401 TI - Late complications after open carpal tunnel decompression. AB - 66 patients (89 hands) were assessed at least 10 months after open carpal tunnel decompression. Tender scars were found in 19% of the hands and 4% were affected by pillar pain. Grip strength was reduced in more than half of the operated hands. Hypo- and anaesthesia in the scar area were affecting 7% but were not considered disabling. In 18% of the cases there was incomplete relief of primary symptoms. PMID- 8732402 TI - Nerve injury following steroid injection for carpal tunnel syndrome. A report of two cases. AB - Two cases of nerve injury are reported following steroid injection as treatment for carpal tunnel syndrome. One caused an ulnar nerve lesion that recovered well. The other caused a more severe median nerve lesion which responded poorly to conservative treatment. Steroid injection for carpal tunnel syndrome is generally safe but nerve injury may occur and is difficult to treat. PMID- 8732403 TI - The value of wrist arthroscopy. An evaluation of 129 cases. AB - In order to evaluate diagnostic and therapeutic wrist arthroscopy we analyzed 129 arthroscopies with a follow-up of at least 6 months. Seventy-seven arthroscopies were performed for therapeutic purposes; 52 arthroscopies were diagnostic. There were diagnostic benefits in 55 arthroscopies (42.5%), therapeutic benefits in 29 arthroscopies (22.5%), combined diagnostic and therapeutic benefits in 39 (30%) and no benefits in six (5%). In 65 cases of the therapeutic group (with preoperative diagnosis) the authors found that the arthroscopy had been worthwhile. For the diagnostic group without a preoperative diagnosis, an arthroscopic diagnosis was made in 44 cases. Complications occurred in two patients: one tendon incision over a Kirschner wire in the therapeutic group and one superficial infection in the diagnostic group. PMID- 8732404 TI - Ulnar buttress arthroplasty for reconstruction after resection of the distal ulna for giant cell tumour. AB - A giant cell tumour of the distal end of the ulna was treated by en bloc resection. The resected distal end of the ulna was replaced by an iliac bone graft, preserving the triangular fibrocartilage complex. The wrist was pain-free and had a full range of motion 6 months postoperatively. PMID- 8732405 TI - A comparison of immediate and staged reconstruction of the dorsum of the hand. AB - The treatment of complex dorsal hand lesions involving skin and subcutaneous tissues, extensor tendons, and bone remains a difficult problem for reconstructive surgeons. Traditional treatment of these defects uses staged reconstruction, first obtaining soft tissue cover and then performing bone and tendon grafts. The purpose of this study was to compare a series of seven patients who underwent staged reconstruction with seven patients who had immediate reconstruction involving primary bone and tendon grafting. All procedures were performed to correct similar severe dorsal hand defects. Patients with immediate reconstruction had a significantly faster return to maximum range of movement (ROM) (214 days compared to 630 days, P = 0.002), significantly fewer operations (2.1 compared to 5.9, P = 0.002) and a greater chance of returning to work (86% compared to 48.2%, P = 0.3) than patients with staged reconstruction. PMID- 8732406 TI - Resurfacing relatively large skin defects of the hand using arterialized venous flaps. AB - Skin flaps nourished by arterial blood entering through the venous network were applied in 12 cases of relatively large skin defects of the hand. The flaps showed remarkable oedema and multiple bullae on their surface from the second to the 14th postoperative day, but partial necrosis of the flap only developed in three cases. The free arterialized venous flap is a useful procedure for resurfacing relatively large skin defects of the hand. PMID- 8732407 TI - Fasciculated intramedullary pinning of metacarpal fractures. AB - We reviewed 20 patients with 23 displaced extraarticular fractures of the distal end of the long metacarpals, treated by fasciculated pinning. At a mean follow-up of 5 years, all the patients were subjectively satisfied with the result. All the patients but one, who fractured four metacarpals, had a normal range of motion. The average grip strength was 43.4 kg for the operated side compared to 43.4 kg for the uninjured hand. Radiologically, the fractured fifth metacarpal had a shortening of 2.2 mm compared to a control group, whereas the volar angulation was 16.6 degrees (13.2 degrees for the control group). The technique of intramedullary fasciculated pinning is a reliable alternative when conservative treatment of fractures of the metacarpal neck has failed or is inappropriate. The procedure provides sufficient stability to allow early mobilization and a good functional result. PMID- 8732408 TI - The posterior interosseous forearm island flap for skin defects in the hand and elbow. A prospective study of 51 cases. AB - The authors' experience of 51 posterior interosseous forearm island flaps is presented. Its main indications were inadequate scar or scar contraction and complex trauma with gross skin loss, either acute or postprimary. The flap healed uneventfully in 45 cases (88%), but in six it was partially or subtotally lost due to necrosis. Late complications included hair growth in the first web and palm, flap redundancy and hypertrophic scar at the donor site. The posterior interosseous island flap produces excellent skin coverage but is difficult to raise and tends to develop oedema and occasionally necrosis. PMID- 8732409 TI - Diabetes and trigger finger. AB - Trigger finger is a stenosing tenovaginitis in which there is a constriction of the annular sheath associated with a nodule in the digital flexor tendon. Clinically this can present as triggering or snapping of the nodule as it passes through the tight constricting tendon sheath. Although many triggering fingers settle spontaneously, and others respond to local anesthetic and steroid injection, others require surgical decompression. The association between diabetes and trigger finger, although not widely known, has been described and forms part of the "diabetic hand syndrome". We studied 100 adult patients in the group that required surgery. Eighteen of the 100 patients were diabetic with six insulin dependent diabetics (IDDM) and 12 non-insulin dependent diabetics (NIDDM). In the current climate of day case surgery this has significant implications both for the management of known diabetics as well as the preoperative detection of those with the condition. PMID- 8732410 TI - A comparison of methods of treatment of pip joint contractures in Dupuytren's disease. PMID- 8732411 TI - Does an ulnar nerve lesion influence the motion of the index finger? AB - Clawing of the ring and little fingers and "rolling" during prehension grip are well-known clinical phenomena of ulnar nerve lesions. In contrast to this the index and middle fingers seem to move normally. We compared the movement of right index fingers in healthy people with the right index fingers of people with an ulnar nerve lesion. The movement was measured using a three-dimensional real time motion analysis system based on ultrasound. The angles of the joints were plotted in a rectangular coordinate system. Statistical analysis of the numerical data showed no difference between the two groups. We trained a neural-network (Learning Vector Quantization) with the data of both groups. The network was able to distinguish between people with and without lesions of the ulnar nerve. We conclude that prehension grip of the index finger is also influenced by paralysis of the ulnar nerve. PMID- 8732412 TI - Excursion of the ulnar nerve at the elbow following epicondylectomy or transposition. AB - We studied the excursion of the ulnar nerve at the elbow following various surgical treatments for ulnar nerve compression in a cadaver model. The ulnar nerve length was measured in various positions of flexion in four situations: the normal anatomical position, following epicondylectomy and following submuscular and subcutaneous transposition. The changes in ulnar nerve length from 0 degrees to 120 degrees of flexion in the anatomical position and after epicondylectomy were significantly less than in the other situations. There was no significant difference in length between the normal anatomical situation and after epicondylectomy. On the theory that the least nerve excursion is better in the treatment of a nerve palsy, we recommend simple decompression or medial epicondylectomy for the treatment of tardy ulnar nerve palsy at the elbow. PMID- 8732413 TI - Glomus tumours of the hand. A retrospective study of 51 cases. AB - The authors report a series of 51 patients with glomus tumours in the hand. The duration of symptoms before treatment averaged 10 years. No one site or finger was more commonly involved. Objective features were limited to a blue discoloration in 29%, a pulp nodule or a nail deformity in 33%. An osseous defect was seen on plain X-ray films in 36%. Diagnosis depended on clinical suspicion in 90%. Careful dissection and complete excision of the tumour almost always offer permanent relief. A direct transungual approach was used in the subungual tumours with only one cosmetic problem. Recurrence of symptoms occurred in only two cases after a pain-free interval of 2 years. PMID- 8732414 TI - Mucormycosis of the upper limb. AB - A rare case of mucormycosis of the upper limb in a patient with AIDS is described. The pathophysiology and natural history of the disease are described along with a classification of its clinical stages. PMID- 8732415 TI - Obstetrical brachial plexus injury in newborn babies delivered by caesarean section. AB - The hand surgeon is frequently involved in the medicolegal assessment of birth palsy cases. Although brachial plexus injury has been reported in newborns delivered by Caesarean section, it is difficult to determine if these cases are due to excessive force when delivering the infant from the uterus or whether the palsy is related to other factors. We have studied our series of 16 cases of obstetrical brachial plexus palsy with special attention to the newborn baby delivered by Caesarean section. We have reviewed the English literature over the last decade and found that birth palsy in newborns delivered by Caesarean section is extremely rare (1% of all birth palsy cases). The differentiation between brachial plexus injury caused by forcible delivery and congenital upper limb palsy from other causes is discussed. PMID- 8732416 TI - Classification of hand anomalies in Apert's syndrome. AB - The literature states that all patients with Apert's syndrome should have complex (osseous) syndactyly of the index, middle and ring fingers. We describe a case of Apert's syndrome with simple syndactyly and recommend extending the classification of hand anomalies in Apert's syndrome. PMID- 8732417 TI - Gantzer's muscle. An anatomical study of the accessory head of the flexor pollicis longus muscle. AB - The relationship of Gantzer's muscle (the accessory head of flexor pollicis longus; FPL) to the median nerve and its anterior interosseous branch is debated. In an anatomical study of 25 limbs, the incidence, origin, insertion, nerve supply and relations of Gantzer's muscle have been documented. The muscle was found in 13 of the 25 (52%) limbs and was supplied by the anterior interosseous nerve. It arose from the medial humeral epicondyle in 85% and had a dual origin from the epicondyle and coronoid process in the rest. Its insertion was to the ulnar part of FPL and its tendon. Gantzer's muscle always lay posterior to both the median and anterior interosseous nerves. Anatomical variations in which Gantzer's muscle may contribute to median nerve compression in pronator syndrome are discussed. PMID- 8732418 TI - Dysfunctional postures of the hand as part of a conversion reaction. AB - A dysfunctional posture of the hand could be due to an anatomical disorder or a conversion reaction. A conversion reaction implies that an unconscious intrapsychic conflict is expressed in a physical dysfunction. Treatment of dysfunctional postures due to conversion reactions is often difficult, and case reports are used to outline management. The need for a multidisciplinary approach is stressed. PMID- 8732419 TI - Florid reactive periostitis of the thumb. AB - A case of florid reactive periostitis of the thumb is presented. This is a rare pseudosarcomatous lesion which generally has a benign radiographic appearance, but on histology it may be mistaken for a malignant process. The clinical behaviour may simulate a malignant tumour but after local excision there is very little propensity for recurrence. Local resection is the recommended treatment. Surgeons and pathologists should consider the possibility of florid reactive periostitis before making a diagnosis of sarcoma in the hand. PMID- 8732420 TI - Recurrent eccrine acrospiroma of the hand. PMID- 8732421 TI - Post-traumatic digital overgrowth. AB - A case is reported of length discrepancy between the two thumbs in a 12-year-old girl following trauma to her left thumb twice before the age of 3 years. The distal phalanx of the left thumb overgrew by 5 mm. There was no functional disability and surgery was requested to improve the appearance. PMID- 8732422 TI - Absorbable sutures in tendon repair. PMID- 8732423 TI - Long-term results of carpal tunnel release. PMID- 8732424 TI - Usefulness of preoperative nerve conduction studies. PMID- 8732425 TI - Histogenesis of vascular tumours. PMID- 8732426 TI - Over-distraction of the radiocarpal and midcarpal joints with external fixation of comminuted distal radial fractures. PMID- 8732427 TI - Positive and negative immunomodulation by opioid peptides. AB - The data that follow review part of the existing evidence concerning the neuroimmune functions mediated by opioid peptides, with particular regard to dual immunomodulatory effects. Limited references to substances other than opioid peptides are included, mainly to emphasize the possible similarities in the mediation of neuroimmune interactions by different informational substances, while the interactions directed from the immune to the nervous system have deliberately been omitted. PMID- 8732428 TI - Induction of ileum muscarinic cholinoceptor signal transduction pathways by rat interferon-gamma. AB - We have recently shown that rat interferon-gamma (IFN gamma) can trigger a cholinergic muscarinic response upon interaction with isolated rat atria, decreasing both the contractile tension and cAMP synthesis, and increasing cGMP production. Because the intestine is rich in cholinergic muscarinic receptors, and IFN gamma-producing cells are normal components of the mucosal immune system, we investigated whether IFN gamma could also modulate the muscarinic cholinergic activity of the intestine. Our results indicate that IFN gamma may interact with ileum cholinergic muscarinic receptors by inhibiting binding of the radiolabelled antagonist quinuclidinyl benzylate (3H-QNB), decreasing cAMP formation and increasing cGMP synthesis. These effects are associated with increased mechanical activity of the isolated ileum strips, and can be blunted by pretreatment of the tissue with atropine. PMID- 8732429 TI - Further evidence on the interaction of muramyl dipeptide with the serotonergic system. AB - The mode of interaction between muramyl dipeptide (MDP), a compound with immunopharmacological activities, and 5-hydroxtryptamine (5-HT, serotonin) was studied in isolated nerve-smooth muscle preparations of the carp stomach. Application of exogenous 5-HT evoked direct smooth muscle contractions; electric neurogenic stimulation evoked twitches due to release of 5-HT from nerve endings. Contractions evoked by a high concentration of 5-HT (3-30 microM) were resistant to atropine and potentiated in the presence of MDP. Isamoltan (5-HTID antagonist) decreased the amplitude of contractions, whereas ketanserin (5-HT2 antagonist) and MDL 72,222 (5-HT3 antagonist) had no effect. The addition of low concentrations (0.1-1.5 microM) of 5-HT did not contract the preparation but caused a decrease in the amplitude of neurogenic twitches, which might be due to the presynaptic inhibition of serotonin release. This effect of 5-HT was not changed by isamoltan or ketanserin, but it was largely reduced in the presence of 5-HT3 antagonists tropisetron and MDL 72,222. This inhibitory effect of 5-HT on twitch amplitude was potentiated by MDP. The interaction of MDP with the serotonergic system thus involved not only potentiation of the postsynaptic effect of higher 5-HT concentrations, which might have been mediated via the 5 HT1 subsystem, but also presynaptic inhibition. MDP enhancement of 5-HT's inhibitory effect, mediated via 5-HT3 receptors, might represent a new feature in mutual 5-HT-MDP interactions. PMID- 8732430 TI - A new assay system detecting antibody production and delayed-type hypersensitivity responses to trinitrophenyl hapten in an individual mouse. AB - A new assay system detecting antibody production and delayed-type hypersensitivity (DTH) responses to trinitrophenyl hapten in an individual mouse (AS-DAD) was established. BALB/c mice were immunized intraperitoneally with varying amounts of 2,4,6-trinitrophenylated sheep red blood cells (TNP-SRBC) on day 0. Venous blood was collected on days 2, 4, 6, 8 and 10. Levels of anti-TNP IgM and IgG serum were assayed by enzyme-linked immunosorbent assay (ELISA). After series of bleeding the mice were challenged with 2,4,6-trinitrobenzene sulfonic acid (TNBS) solution in the footpad on day 14. Footpad swelling was measured 24 or 48 h after the challenge. Peak responses of the anti-TNP IgM and IgG production were detected 4 or 6 days after the immunization with 10(9) TNP SRBC. Maximum DTH response was also observed with 10(9) TNP-SRBC 24 h after the challenge on day 14. The antibody and DTH responses were also induced in other normal inbred strains such as C3H/He and DBA/1 but not BALB/c nu/nu mice. To evaluate AS-DAD in immunopharmacological studies, various immunomodulating agents were examined in BALB/c mice by subcutaneous administration on days 0, 1, 2 and 3. Cyclosporin or cyclophosphamide at 100 mg/kg/day completely inhibited not only the anti-TNP IgM and IgG production but also the TNP-specific DTH response. Prednisolone at 0.5 mg/kg/day had no significant effect on the IgM and IgG production, whereas it inhibited the TNP-specific DTH response. Interestingly, histamine-added mouse gamma-globulin at 150 MG/kg/day clearly enhanced the anti TNP IgM and IgG production, while it showed a suppressive effect on the TNP specific DTH response. Levamisole at 5.0 mg/KG/day showed suppressive effects on the anti-TNP IgG production without affecting the IgM production and the DTH response. These results suggest that AS-DAD is useful for evaluating the immunopharmacological action of various agents. PMID- 8732431 TI - Fu-Ling, a Chinese herbal drug, modulates cytokine secretion by human peripheral blood monocytes. AB - Fu-Ling, the sclederma of Poria cocos (Schw.) Wolf, has long been used as a sedative and diuretic in traditional Chinese herbal medicine. Our study demonstrated that the substances extracted from Fu-Ling by 50% hot ethanol significantly augmented the secretion of interleukins IL-1 beta and IL-6 6 h after in vitro cultivation of human peripheral blood monocytes. The augmented effect was dose dependent. Tumour necrosis factor-alpha (TNF-alpha) secretion was also increased as the cells were treated with 0.4 mg/ml or higher doses of Fu Ling extract. By contrast, Fu-Ling extract significantly suppressed the secretion of transforming growth factor-beta (TGF-beta) 3 h after the in vitro drug treatment. The suppressive effect was shown at doses as low as 0.2 mg/ml of Fu Ling extract. Since Fu-Ling extract enhanced the secretion of immune stimulators (IL-1 beta, IL-6 and TNF-alpha) but suppressed the secretion of an immune suppressor (TGF-beta), the substance in 50% hot ethanol extract of Fu-Ling might have potentiated the immune response. Fu-Ling extract was further fractionated by reverse-phase column chromatography and high-performance liquid chromatography (HPLC). The components showing activity in modulating the cytokine secretion were relatively high in hydrophobicity. PMID- 8732432 TI - Inhibition of re-expression of surface CD4, but not CD8, on activated human T lymphocytes by the immunosuppressive drugs dexamethasone and cyclosporine A: correlation with inhibition of proliferation. AB - Our objective was to study the re-expression of CD4 and CD8 on activated human T lymphocytes in the presence of dexamethasone and cyclosporine A at physiologically relevant concentrations. Peripheral blood mononuclear cells from 12 healthy volunteers were cultured for up to 72 h in the presence and absence of phytohaemagglutinin A (PHA) and dexamethasone and cyclosporine A at a range of concentrations. Expression of surface CD3, CD4 and CD8 and the activation marker CD25 was quantified on T-lymphocytes using flow cytometry. T-lymphocytes cultured with PHA showed significant (P < 0.01) reduction in the expression of surface CD3, CD4 and CD8 compared with cells cultured in medium alone, which was apparent as early as 0.5 h after PHA exposure. The reduction in the expression of both CD4 and CD8 in the first 0.5 h after exposure correlated with that of CD3 (P < 0.02). Expression of CD4 and CD8 subsequently increased again, so that by 48 h after exposure CD4 expression was no longer significantly different to that of cells cultured in medium alone. CD8 expression recovered later, but within 72 h. Both dexamethasone and cyclosporine A, when added at the commencement of the cultures, significantly inhibited, in a concentrated-dependent (10(-6)-10(-9) mol/l) fashion, the re-expression of CD4, but not CD8, on the PHA-stimulated T lymphocytes measured after 48 h. In contrast, CD25 expression on both CD4 and CD8 T-lymphocytes was inhibited by both drugs to an equivalent extent over the same range of concentrations. At this time point, the degree of inhibition of CD4 re expression by both drugs correlated significantly with the degree of inhibition of CD25 expression. We concluded that activation of T-lymphocytes results in rapid down-modulation of surface CD3, CD4 and CD8 followed by re-expression which is inhibited in the case of CD4, but not CD8, by dexamethasone and cyclosporine A. Since CD4 is thought to co-operate in T-lymphocyte antigen recognition by the CD3-antigen receptor complex, this effect may contribute to the immunosuppressive effect of these drugs on CD4 T-lymphocytes. PMID- 8732433 TI - Delta-9-tetrahydrocannabinol suppresses tumor necrosis factor alpha maturation and secretion but not its transcription in mouse macrophages. AB - Various in vitro studies have shown that delta-9-tetrahydrocannabinol (THC), the major psychoactive component of marijuana, has a variety of inhibitory effects on immune functions including effects on macrophages. The present studies have examined the mechanism of THC's effects on tumor necrosis factor alpha (TNF alpha), a major macrophage-produced cytokine and an important mediator involved in cytokine networks and in host defense mechanisms. Exposure of macrophages to medium containing THC has resulted in low levels of soluble TNF-alpha protein and reduced TNF-alpha bioactivity in the culture supernatant. However, THC did not inhibit the levels of LPS-induced TNF-alpha mRNA and intracellular TNF-alpha precursor protein, had only a weak effect on expression of membrane-bound TNF alpha, but suppressed TNF-alpha maturation/secretion by macrophages. The higher the THC concentration in the medium during TNF-alpha induction, the greater the amount of intracellular TNF-alpha precursors that accumulated in the activated macrophages and the less mature TNF-alpha was released from the cells. Data suggest that TNF-alpha production by macrophages was altered greatly by exposure to THC at the levels of TNF-alpha precursor maturation and secretion. PMID- 8732434 TI - RU 41 740 (Biostim) and IL-4, or IL-13, have opposite effects on CD14, CD23, HLA DR and HLA-DQ on monocytes. AB - RU 41 740 (Biostim) is a glycoprotein extract obtained from Klebsiella pneumoniae. Its immunostimulating properties on monocytes have been established in vivo and in vitro. To confirm its spectrum of action at molecular level we studied its role on the modulation of four molecules involved in antigen presentation (HLA-DR, HLA-DQ), uptake of endotoxin (CD14) and activation (CD23). These four molecules are known to be modulated by interleukins IL-4 and IL-13. We found that HLA-DR, HLA-DQ, CD14 and CD23 were differentially regulated by biostim and IL-4 or IL-13. Surprisingly, Biostim inhibited the IL-4 or IL-13-induced expression of CD23, HLA-DQ and HLA-DR, while it did not have any action on these molecules by itself. We therefore hypothesize that Biostim, through the action on its receptor, could interact with the IL-4 receptor and IL-13 receptor and/or inhibit the IL-4 and IL-13 receptor transducing signal. PMID- 8732435 TI - Effect of nedocromil sodium and cromoline sodium on atopic basophil function. AB - Nedocromil sodium (NS) is a new drug for the treatment of bronchial asthma. In the present study, we examined the effect of NS on basophil histamine release stimulated by anti-IgE, anti-IgE+IL-3 (interleukin-3) or ryegrass. The effect of cromoline Na (CrS) on histamine release from basophils to the same stimuli was compared. NS did not affect histamine release from basophils following anti-IgE alone or together with IL-3, but augmented the release following ryegrass. CrS did not affect histamine release after the same three stimuli. We concluded that NS and CrS do not affect bronchial asthma through direct inhibition of histamine release from basophils. Their action may be via an indirect effect on histamine releasing factor. PMID- 8732436 TI - Comparative evaluation of in vitro and in vivo immunosuppressive potential of cyclosporin G with cyclosporin A and FK-506. AB - Cyclosporin G (CsG), a promising cyclosporin A (CsA) analogue, was examined and compared with two reference immunosuppressive drugs: CsA and FK-506, regarding their inhibitory effects on different lymphocyte activation pathways as well as on graft-versus-host reaction (GvHR) across differences at major or minor histocompatibility loci. The results showed that, at different concentrations, CsG efficiently inhibited proliferation induced by alloantigens (mixed lymphocyte culture), mitogens (concanavalin A, pokeweed mitogen) and the combination of phorbol myristate acetate + ionomycin, to the same extent as observed with CsA and FK-506. It was also shown that CsG exhibited the same strong inhibitory effects as the two other immunosuppressants upon stimulation triggered by viral (MLs-1a) or bacterial (staphylococcal enterotoxin B) superantigen. Determination of IL-2 activity in the supernatant of MLC also confirmed similar strong inhibitory effects, exerted by CsG compared to CsA and FK-506. In systemic and local GvHR across major or minor histocompatibility barriers, CsG as well as CsA and FK-506 presented an equivalent immunosuppressive potential. In conclusion, from various experiments involving different modes of activation, it was shown that CsG was as strongly immunosuppressive as CsA and FK-506. PMID- 8732437 TI - IL-6-soluble IL-6 receptor complex inhibits the proliferation of dermal fibroblasts. AB - A number of investigators has reported that there is increased production of interleukin-6 (IL-6) by fibroblasts and monocytes from the patients with systemic sclerosis (SS). However, the precise role of IL-6 in the pathogenesis of SS remains unclear. On the basis of our previous study showing that the complex of IL-6 and soluble IL-6 receptor (sIL-6R) could induce synovial fibroblast proliferation, we examined whether the IL-6-sL-6R complex could induce the proliferation of normal dermal fibroblastic cells (DF). IL-6 suppressed DF proliferation, and, in the presence of sIL-6R, dose-dependently showed much stronger suppressive effects on DF proliferation. This suppression was completely blocked by either anti-IL-6 or anti-sIL-6R antibody. Furthermore, the IL-6-sIL-6R complex significantly suppressed IL-1 beta-, TNF alpha- and PDGF-AA-induced DF proliferation. These lines of evidence suggest that the IL-6-sIL-6R complex may have potential as a useful agent for the treatment of SS. PMID- 8732438 TI - Pharmacology and pharmacokinetics of citalopram and other SSRIs. AB - Citalopram together with fluoxetine, fluvoxamine, paroxetine and sertraline belong to the group of SSRIs, so named because of their pharmacological action as selective serotonin reuptake inhibitors in rat brain synaptosomes-their potency of inhibiting noradrenaline uptake is low and, from a clinical point of view, irrelevant. In contrast to classical tricyclic antidepressants and some antipsychotics, the SSRIs have little affinity for the dopamine D2 receptors, 5 HT1A and 5-HT2A receptors, alpha 1-receptors, beta-receptors, muscarinic receptors and histamine H1 receptors. Several authors have examined whether SSRIs are ligands of other receptors, including the 5-HT3-5-HT7 receptors and their subtypes, and the NMDA receptor, and whether chronic treatment with SSRIs modifies the affinity and binding capacity of these receptors. The SSRIs differ by their pharmacokinetics and pharmacogenetics of metabolism and by their cytochrome P450 isozyme inhibition properties. Some situations are presented in which plasma level monitoring of SSRIs is recommended, despite the lack of clearly defined "therapeutic windows'. PMID- 8732439 TI - Does selectivity matter? AB - An important aim of drug development has been to retain efficacy while reducing side effects and toxicity. It is now clear that selective serotonin reuptake inhibitors (SSRIs) are more effective than noradrenaline reuptake inhibitors (NARIs) in the treatment of obsessional compulsive disorder, and obsessional features in depression predict a responsivity to SSRIs. Furthermore, panic disorder and depression may be more responsive to SSRIs than NARIs. However, there is evidence that inhibition of both sites produces slightly greater efficacy and the addition of an NARI has been reported to potentiate the antidepressant effect of SSRIs. Tricyclic noradrenaline-serotonin uptake inhibitors have many other pharmacological properties, which probably relate to high rates of side effects and cardiotoxicity. Whether in practice these features reduce compliance or increase deaths from suicide is debatable, but it seems wrong to subject patients to burdensome side effects. Certainly, in overdose, older non-selective antidepressants are far more likely to kill than SSRIs. SSRIs have their own side effects, such as nausea and anorgasmia, due to their potency at the uptake site, but these are reversible and sometimes treatable. Clearly, selectivity for specific uptake sites matters for efficacy, and selectivity for uptake over other sites matters for tolerability and safety. PMID- 8732440 TI - Serotonin: current understanding and the way forward. AB - Serotonin is a key neurotransmitter in the central nervous system. In recent years its role in the pathophysiology and pharmacology of depression has been extensively investigated. Neuroendocrine studies suggest decreased serotonergic responsivity in patients with major depression. The fact that the majority of patients become depressed in a setting of stressful life events may indicate that the neurotransmitter disturbance occurs as part of the stress response in certain vulnerable individuals. Effective treatment of depression results in normalization of this disturbance. Drugs which selectively block the reuptake of serotonin are effective antidepressants with a low incidence of side effects. PMID- 8732441 TI - Are pharmacokinetic drug interactions with the SSRIs an issue? AB - The development of the selective serotonin reuptake inhibitors (SSRIs) began 20 years ago, around the time when it was discovered that the cytochrome P450 system consists of multiple drug-metabolizing enzymes. There are 5-10 important drug metabolizing P450 enzymes in the human liver, and their relationship with SSRIs has been studied intensively during the last 5 years. Thus, among the SSRIs, fluvoxamine is the only very potent inhibitor of cytochrome P4501A2 (CYP1A2). All of the SSRIs inhibit CYP2D6 ('sparteine/debrisoquine oxygenase') but fluoxetine and paroxetine are clearly the most potent in this regard. Fluoxetine and fluvoxamine are moderate inhibitors of CYP2C19 ('S-mephenytoinhydroxylase'), and fluvoxamine might also be a moderate inhibitor of CYP2C9. Thus, although much still has to be learned about SSRIs and cytochrome P450, it seems that citalopram and sertraline have the most favourable profile in relation to drug interactions. PMID- 8732442 TI - The antidepressant efficacy of citalopram. AB - Depression is a long-term illness requiring psychopharmacological treatment over long periods of time. Treatments need to be effective and safe, but in an illness requiring long-term medication, acceptability to patients must also be addressed. Data from eight placebo-controlled studies addressing short-term efficacy and two placebo-controlled studies of long-term (6 months) treatment have demonstrated the antidepressant efficacy of citalopram. Fourteen studies comparing citalopram with a reference antidepressant have provided supporting evidence of efficacy. In all, 3905 patients have been included in clinical efficacy studies, in which 2579 patients have been treated with citalopram, 486 with placebo and 840 with a reference antidepressant. The recommended dose of 20 mg appears appropriate for most patients, with the option of raising the dose in non-responders, particularly if they have more severe depression. PMID- 8732443 TI - The safety and tolerability of citalopram. AB - Since citalopram was first approved in 1989, it has been prescribed to an estimated > 600 000 patients. An integrated safety database has been prepared, including data from 3107 patients from 24 clinical trials. In placebo-controlled trials, nausea, dry mouth, somnolence, increased sweating, tremor, diarrhoea, and ejaculation failure, mostly of mild to moderate severity, occurred significantly (p < 0.05) more frequently with citalopram. The excess incidence of these events over placebo was always less than 10%. In pooled comparative studies, citalopram's tolerability profile was similar to that of other selective serotonin reuptake inhibitors (SSRIs) and superior to that of tricyclic antidepressants (TCAs). Spontaneous adverse event reports arising from clinical use have confirmed the safety profile defined during the trials programme. Specific monitoring of all serious adverse events from around 10 000 patients receiving citalopram in clinical trials (including small open studies) has indicated a low potential for convulsions and extrapyramidal effects. There is no evidence of withdrawal phenomena on abrupt discontinuation, no clinically relevant effects on cardiac or laboratory parameters, and little or no effect on psychomotor function. When taken in overdose alone, citalopram appears to have a relatively wide margin of safety. Citalopram has been well tolerated in both short- and long-term use, and the profile seen in trials has been confirmed in the clinic. PMID- 8732444 TI - Scandinavian experience with citalopram in the elderly. AB - Depression is a common disorder among the elderly. Its prevalence is estimated at 12-15%. A multifactorial genesis must be considered. Depression in the elderly often manifests itself in an atypical way, with a somatized clinical picture. As tricyclics are generally too toxic for elderly people, the use of selective serotonin reuptake inhibitors (SSRIs) must be considered in this patient group. Citalopram is an SSRI drug with an attractive pharmacokinetic profile, i.e. few side effects and a low potential for interaction with other drugs. In two inter Nordic studies, this drug significantly improved emotional disturbances in patients with dementia disorders. Significant improvement, with a response rate of 53%, was also seen in elderly people with depression as assessed using the Hamilton Depression Rating Scale. Sixty-six per cent of these patients also suffered from physical illness and had other drug treatment. The drug was well tolerated. PMID- 8732445 TI - Effective management of depression today: report from an interactive workshop. PMID- 8732446 TI - Thyroid peroxidase: experimental and clinical integration. AB - Since TPO plays a cardinal role in regulating many cellular processes of thyroid hormone biosynthesis in thyrocytes, it is important to present a summary of the impact that TPO research on the basis of molecular structure has had on our understanding of thyroid diseases. This review has therefore been written to highlight the biochemical, molecular biological, immunological and clinical aspects of TPO in thyroid follicular cells. As for hormonogenesis, further details of the properties of the superoxide generating system remain unknown, but based on decisive evidence showing what active iodinating species are at initial and principal stages, a novel concept of the mechanism of two electron oxidation has been put forth. Recent progress in studies of analyses of TPO gene structure has also provided stimuli for renewed investigation into a much wider field of thyroidology, including thyroiditogenesis. PMID- 8732447 TI - Growth of differentiation: determination by FSH of the action of insulin-like growth factor-I in cultured rat granulosa cells. AB - Insulin-like growth factor-I (IGF-I) is a potent mitogen in many cell systems. In cultured rat granulosa cells, however, IGF-I is known to be an inducer of differentiation. The present study was conducted to identify the factor which determines the direction of IGF-I action: either DNA synthesis or LH receptor expression. When granulosa cells were incubated with IGF-I in the presence of various concentrations of follicle-stimulating hormone (FSH), DNA synthesis as assessed by [3H]thymidine incorporation was increased only in the presence of low doses of FSH. The stimulatory effect of FSH on DNA synthesis was observed in a very narrow range of FSH concentration between 2 and 10 ng/ml. At higher concentrations, FSH had little effect on DNA synthesis but instead induced expression of receptors for luteinizing hormone (LH), a marker of granulosa cell differentiation. At 5 ng/ml, FSH elicited maximal stimulation of DNA synthesis and simultaneously induced LH receptor expression to some extent. In these cells, DNA synthesis peaked at 36 h but expression of LH receptor occurred later than 36 h, peaking at 60 h. The ability of IGF-I to stimulate DNA synthesis was enhanced by the long term pretreatment with FSH: when FSH was added from the beginning and IGF-I was added after 36 h or later, IGF-I-mediated DNA synthesis was approximately twice as great, and was accompanied by a two-fold increase in the number of bromodeoxyuridine-labeled nuclei. Under these conditions, LH receptor expression was reduced to approximately 50%. Finally when cells were incubated for 12 h with or without FSH, washed extensively with the medium and then IGF-I was added, DNA synthesis was augmented only in FSH-primed cells. Forskolin, an activator of adenylate cyclase, reproduced the effect of FSH. These results indicate that, in the presence of FSH, IGF-I has the ability to induce both DNA synthesis and differentiation and that FSH determines the action of IGF-I on promotion of either growth or differentiation. Furthermore, priming with FSH renders granulosa cells responsive to IGF-I in terms of DNA synthesis. PMID- 8732448 TI - A family of multiple endocrine neoplasia type 2A: genetic analysis and clinical features. AB - Since a heterozygous missense mutation of the RET proto-oncogene in the germline was found to cause multiple endocrine neoplasia type 2A (MEN 2A) in 1993, some 20 different mutations of this gene have been identified in MEN 2A kindreds. We report an MEN 2A family in which serine (AGC) substitutes for cysteine (TGC) at codon 618 in exon 10 of the RET proto-oncogene. The mutation was identified by sequencing PCR products of exons 10 and 11 in the proband. Since this mutation results in creation of a new cleavage site for Alu I restriction enzyme, most of the other members of the family were screened by digestion of the PCR product of exon 10 with this enzyme. Eleven of 20 subjects across four generations examined have the mutation of the RET proto-oncogene, and all of the adult gene carriers except one woman had MTC. Characteristics of this family are 1) pheochromocytoma has been found in only the proband, 2) no obvious hyperparathyroidism has been observed, and 3) the prognosis is favorable, with nobody dying of MEN 2A itself. Genetic analysis of MEN 2A is definitely useful and essential for screening of a MEN 2A family. It is very important to accumulate cases with MEN 2A and investigate the phenotype and the prognosis in each mutation. PMID- 8732449 TI - The relationship between serum levels of interleukin-6 and thyroid hormone during the follow-up study in children with nonthyroidal illness: marked inverse correlation in Kawasaki and infectious disease. AB - Our previous study demonstrated that the serum IL-6 correlated inversely with serum T3 and the T3/T4 ratio in children with acute respiratory infection during the acute phase of illness. To investigate whether serum IL-6 inversely correlates with serum thyroid hormone during not only the active stage of the disease but also in the follow-up period in nonthyroidal illness, we measured serum levels of IL-6, T3 and T4 in 31 children from the acute to the convalescent phase. They were divided into 3 groups; 7 patients with Kawasaki disease, 16 patients with infectious disease and 8 patients with non-inflammatory disease. In the follow-up of patients with Kawasaki disease, a marked inverse relationship was observed between serum IL-6 and T3 (r = -0.844, P < 0.001) or the T3/T4 ratio (r = -0.863, P < 0.001). Serum T4 showed a weak but significant negative correlation with serum IL-6 (r = -0.474, P = 0.035) only in this situation. There was also a significant negative correlation between serum IL-6 and T3 (r = 0.582, P < 0.001) or the T3/T4 ratio (r = -0.660, P < 0.001) during the follow-up of children with infectious disease. In the follow-up study of patients with non inflammatory disease, however, no significant relationships were observed between serum thyroid hormones and IL-6. IL-6 may be one important factor involved in the decrease in the serum T3 level and the T3/T4 ratio in patients with nonthyroidal illness particularly characterized by strong inflammation and activation of the immune system as observed in Kawasaki and infectious disease. PMID- 8732450 TI - Regulation of immunoreactive activin A secretion from cultured rat anterior pituitary cells. AB - Immunoreactive activin A (ir-activin A) release from cultured rat anterior pituitary cells was examined by measuring ir-activin A in culture medium by a specific radioimmunoassay. Ir-activin A release into the medium increased over 1 18 days, and reached a maximal level at 12-15 days. The basal levels of ir activin A in the culture media were 0.70 +/- 0.10 (mean +/- SD), 1.30 +/- 0.36 and 1.83 +/- 0.44 ng/10(6) cells, when cultured for 6 days with 0, 2 and 10% fetal calf serum, respectively. LHRH induced an approximate 1.4-fold increase in ir-activin A release in contrast to a 40-60% inhibition with FSH, but LH did not affect the activin A release. In the presence of 12-o-tetradecanoylphorbol acetate (TPA), iractivin A release was enhanced, but no significant effect was induced by forskolin. Activin A was distinctly immunostained in cultured rat anterior pituitary cells. These results suggested that activin A release from the pituitary is modified by FSH and LHRH, and that the activation of protein kinase C may be involved in the action of LHRH. PMID- 8732451 TI - Prolactin-dependent growth and gamma-casein gene expression in Ba/F3 cells transfected with a long form of mouse mammary prolactin receptor. AB - Complementary DNA (cDNA) encoding a long form of prolactin receptor (PRL-RL) was cloned from mouse mammary gland by PCR using primers designed from the noncoding regions of previously reported rat ovarian PRL-RL cDNA. The nucleotide sequence encoding the extracellular and transmembrane domains of PRL-RL is completely identical to that of short forms of mouse PRL-R. The amino acid sequence deduced from the nucleotide sequence of mouse PRL-RL is 91% identical to that of rat PRL RL. To address the question of whether or not the cloned mouse PRL-RL cDNA encodes a functional PRL-RL we transfected Ba/F3 IL-3-dependent murine pro-B lymphoid cells with the cDNA. By culturing the transfected cells in a medium which contained PRL in place of IL-3, we selected 5 PRL-dependent clones. All of these PRL-dependent clones, BaF/PD cells, expressed PRL-RL mRNA. In addition, BaF/PD cells expressed mammary-specific gamma-casein mRNA in response to PRL and dexamethasone. Based on these results, it was concluded that the mouse mammary PRL-RL cDNA cloned in this study is functionally active in mediating both PRL dependent growth and mammary-specific gene expression. PMID- 8732452 TI - Differential expression of dipeptidyl peptidase IV (CD26) and thyroid peroxidase in neoplastic thyroid tissues. AB - This paper reports the results of research to examine the possibility of using the differential expression of two enzymes, dipeptidyl peptidase IV (DPPIV/CD26, EC: 3.4.14.5) and thyroid peroxidase (TPO, EC: 1.11.1.7), as histochemical markers histopathologically to diagnose thyroid carcinomas. The research is based on previous reports that DPPIV/CD26 is overexpressed in differentiated thyroid carcinoma tissues, and that TPO activity is very low in thyroid carcinoma tissues. Differential expression of the two enzymes in 32 thyroid tissues of various thyroid diseases was studied by Northern blot analysis and histochemical analysis. On Northern blot analyses, all 14 differentiated thyroid carcinomas (11 papillary carcinomas and 3 follicular carcinomas) overexpressed DPPIV/CD26 mRNA, whereas all 17 benign thyroid tissues (4 normal thyroid tissues, 4 Graves' diseases, 2 adenomatous goiters and 7 follicular adenomas) showed faint mRNA expression of DPPIV/CD26. All 17 benign thyroid tissues expressed high levels of TPO mRNA, whereas all 11 papillary carcinomas strongly underexpressed TPO mRNA. Histochemically, all 17 benign tissues were DPPIV/CD26 negative and strongly TPO positive, while all 11 papillary carcinomas were strongly DPPIV/CD26 positive and TPO negative. Two of 3 follicular carcinomas were histochemically positive for the two enzymes. A medullary carcinoma did not show any mRNA expression of either enzyme. These results suggest that the differential expression of these two enzymes can be applied to study the thyroid tumorigenesis. PMID- 8732453 TI - The change in 123I-uptake between 3- and 24-hours is useful in predicting early response to methimazole in patients with Graves' disease. AB - Some patients with Graves' disease respond well to anti-thyroid drug treatment but others do not. Factors determining the patient's responsiveness to the medical treatment are unclear, but the intrathyroidal iodine pool is believed one of the important factors. In this study, we found that delta radioactive iodine uptake (RAIU) (RAIU at 24 h-RAIU at 3 h) is useful in predicting early response to treatment with methimazole (MMI). Among 32 patients with Graves' disease, who were given 30 mg MMI as an initial dose, 11 patients responded quickly to MMI treatment. Within one month, serum free T4 levels decreased to below the normal range in 6 patients (< 10.3 pmol/L) or decreased from beyond the highest level of the assay (> 125 pmol/L) to the normal range in 5 patients. When these rapid responders (group A) were compared with the remaining 21 patients who showed a more gradual response to MMI-treatment (group B), a different pattern of 123I thyroid uptake was noted. RAIU at 3 h was significantly higher in group A than in group B, while RAIU at 24 h was similar in the two groups. As a result, rapid responders had a significantly lower delta RAIU value than gradual responders ( 0.7 +/- 8.4% in group A, 14.2 +/- 8.2 in group B, P < 0.01). No significant difference was found between the two groups in various pre-treatment parameters such as severity and duration of thyrotoxicosis, the titer of TSH receptor antibodies (TRAb), frequency of positive antithyroglobulin antibodies (TGHA), urinary excretion of iodine and thyroid volume. The incidence of positive antithyroid microsomal antibodies (MCHA) was higher in group A than in group B, and thyroid ultrasonography showed a tendency to low echogenicity in group A. delta RAIU was negatively correlated with the reduction in the serum free T4 level during the first two weeks after MMI-treatment was initiated (r = -0.60, P < 0.01). Moreover, delta RAIU correlated positively with the biological half-life of the intrathyroidal iodine, calculated in a different series of 24 patients with Graves' disease who received radioisotope treatment (r = 0.54, P < 0.01). The low delta RAIU value is considered to reflect the rapid turnover of the intrathyroidal iodine, and may be related to the small intrathyroidal iodine pool. delta RAIU is useful in predicting early responsiveness of patients with Graves' disease to MMI-treatment. PMID- 8732454 TI - Increased serum human granulocyte colony-stimulating factor (G-CSF) levels following intravenous infusion of high-dose methylprednisolone. AB - Serum granulocyte colony-stimulating factor (G-CSF) level were measured for 24 h in 6 patients with hematological disorders and two patients with Graves' disease associated with malignant exophthalmos after iv infusion of high-dose methylprednisolone (HDMP) (20 mg/kg BW) for 2 h starting at 0900 h. Saline solution (500 ml) was infused iv for 2 h as a control on two days before starting the HDMP therapy. Serum G-CSF levels increased with the mean peak values at 4 h after the HDMP therapy (mean +/- SD: 488.1 +/- 125.8 pg/ml vs. saline control 74.4 +/- 21.9 pg/ml, P < 0.01). Circulating absolute neutrophil counts (ANC) also increased with the mean peak values at 24 h after the HDMP (6,057 +/- 1,460 / microliters vs. saline control 2,007 +/- 390 / microliters, P < 0.025). These findings indicate that glucocorticoid has a stimulating effect on G-CSF release and that the increased G-CSF release could be involved, at least partly, in neutrophilia induced by glucocorticoid. PMID- 8732455 TI - Leydig cell tumor of the testis: analysis of testosterone production and secretion by three-dimensional histoculture. AB - We treated an 11-year-old boy with a testicular Leydig cell tumor. We analyzed the testosterone production of this tumor by immunolocalization of steroidogenic enzymes and in vitro three-dimensional histoculture. Spermatic venous blood from the tumor bearing testis had noticeably high concentrations of testosterone and androstenedione. The tumor had the characteristic ultrastructural features of steroid producing cells and was immunoreactive for P450scc (side chain cleavage), 3 beta HSD (hydroxysteroid dehydrogenase) and P450c17 (17 alpha-hydroxylase). Three-dimensional collagengel-supported histoculture demonstrated that the tumor tissue in the culture maintained its histologic architecture, expression of steroidogenic enzymes, and secretion of testosterone into the medium for up to 7 days in culture. Histoculture preserved in vitro testosterone production in this case of testicular Leydig cell tumor. PMID- 8732456 TI - Incidentally discovered adrenocortical adenomas are not fully nonfunctioning: immunohistochemical and dispersed adrenocortical cell study. AB - To clarify the precise function of incidentally discovered adrenocortical adenoma, immunohistochemical and dispersed adrenal cell studies were performed. We have recently seen five patients with so-called nonfunctioning adrenocortical adenoma. Diurnal variation in plasma cortisol and suppression of plasma cortisol and urine 17-hydroxycorticosteroids in response to dexamethasone administration revealed adrenocortical function within normal limits in all cases, and no signs or symptoms of adrenal steroid hormone excess were evident. Since a high uptake of iodomethylnorcholesterol was recognized in each adrenal mass, it was supposed that these adrenal tumors produced steroid hormone to a certain extent, and each patient received unilateral adrenalectomy. P450c17, a key enzyme involved in cortisol production, was expressed in the tumor region in all cases in an immunohistochemical study. Upon in vitro steroidogenesis with dispersed adrenal cells in two cases, all steroid hormones measured except for aldosterone (progesterone, 17 alpha-hydroxyprogesterone, pregnenolone, 17 alpha hydroxypregnenolone, 11-deoxycortisol, cortisol, 11-deoxycorticosterone, corticosterone, 18-hydroxydeoxycorticosterone, dehydroepiandrosterone and androstenedione) were produced in a culture medium. The results indicated that these tumors possessed the capacity for cortisol production, which was in agreement with the results of an iodomethyl-norcholesterol scintigraphy. All patients with mild hypertension or diabetes mellitus had no signs or symptoms of steroid hormone excess, but they could potentially develop a steroid excess syndrome such as Cushing's syndrome in the future. PMID- 8732457 TI - Recombinant human growth hormone acts on intermediate-sized follicles and rescues growing follicles from atresia. AB - In order to clarify the role of ovarian function, recombinant human GH (rhGH) was administered to adult spontaneous dwarf rats (SDR) for 14 days. The SDR were sacrificed on the day of estrus and the effects of rhGH on ovarian weight, follicular population and the viability estimated by 5-bromodeoxyuridine (BrdU) incorporation, were studied. RhGH administration caused a significant gain in body weight of the SDR, and the ovarian weight of the GH-injected SDR showed a significant increase in comparison with saline injected SDR. The total population of follicles was significantly greater in the GH-treated group than in the control, and the difference was attributed to the increase in the number of large follicles more than 500 micrograms. Total uptake of BrdU into granulosa cells was significantly higher in GH-treated rats than in the control, and, in particular, the intermediate-sized follicles (300-500 micrograms) of the GH-treated rats showed signs of high incorporation of BrdU. These results suggest that hGH acts on intermediate follicles, stimulates their viability and increases the number of growing follicles. PMID- 8732458 TI - Prolactin causes the dissociation of prolactin from plasma membrane receptor in lactating mouse mammary cell: action of high prolactin concentration. AB - In order to characterize the dissociation of receptor-bound prolactin (PRL) by mammary cells, cells were prepared from lactating mice by collagenase digestion and loaded with PRL. The dissociation reaction was performed in the presence of PRL. In the concentration range 1 ng/ml-1 micrograms/ml examined, PRL at higher than 10 ng/ml accelerated the dissociation of PRL in an concentration-dependent manner. The action of PRL on dissociation was completed within a short period. At the end of the 1 h-incubation period, the dissociation rate constant (k-1) was about 2 times larger in the presence of 1 microgram/ml of PRL than in its absence. The action of PRL occurred predominantly at the level of the plasma membrane receptor. Mammary cells with greater PRL-binding capacities had larger k 1 in response to PRL. The present data showed that the dissociation of PRL from the receptor was influenced by the concentration or PRL and by the PRL-binding capacity of the cell. The rate of PRL-receptor interaction is expressed by the equation of k-1 (PRL-bound receptors). It is probable that the exceedingly high levels of PRL, the PRL-receptor interaction occurs more frequently in the presence of PRL-dependent dissociation than in its absence. PMID- 8732459 TI - Involvement of arginine vasopressin and renal sodium handling in pathogenesis of hyponatremia in elderly patients. AB - The present study was undertaken to determine the pathophysiological role of arginine vasopressin (AVP) in elderly patients with hyponatremia, and the efficacy of fludrocortisone acetate in treating their hyponatremia. Eleven hospitalized patients aged 65 years or older whose serum sodium levels were less than 130 mEq/l were examined. The hyponatremic patients included two groups of patients: syndrome of inappropriate secretion of antidiuretic hormone (SIADH) and central salt-wasting syndrome. And 24 healthy, young subjects aged 20 to 34 years, and 24 healthy, elderly subjects age 65 to 80 years were recruited by community announcement. The elderly subjects had decreased urinary concentrating ability and exaggerated response of AVP secretion to osmotic and nonosmotic stimuli, as compared to the young subjects. All the patients had hyponatremia, with the exaggerated urinary loss of Na. Plasma AVP levels were elevated despite hypoosmolality in all the 2 groups of hyponatremic, elderly patients. Plasma renin activity and plasma aldosterone concentrations were low in the patients with SIADH and central salt-wasting syndrome. Fludrocortisone acetate therapy was effective in the patients with central salt-wasting syndrome and 3 patients with SIADH whose hyponatremia remained unchanged after water restriction. Water restriction therapy normalized serum Na levels in only 3 patients with SIADH. These results indicate that AVP is involved in the mechanism for hyponatremia in the elderly patients with SIADH and central salt-wasting syndrome. Severe hyponatremia associated with SIADH and central salt-wasting syndrome responds well to mineralocorticoid therapy. Both the secretion of AVP and renal sodium handling may be involved in the mechanism of action of the disorders. The diagnostic criteria for SIADH in the elderly patients may have to be reevaluated and should be considered to indicate fludrocortisone acetate therapy. PMID- 8732460 TI - Absence of mutations at codon 768 of the RET proto-oncogene in sporadic and hereditary pheochromocytomas. AB - Sixteen sporadic pheochromocytomas, 3 pheochromocytomas in neurofibromatosis 1, and 4 pheochromocytomas in multiple endocrine neoplasia (MEN) 2A or 2B were screened for mutations at codon 768 of the RET proto-oncogene by AluI digestion of polymerase chain reaction PCR products and mutations in exon 13 by PCR-single strand conformation polymorphism (SSCP) analysis. Although mutations at codon 768 (GAG --> GAC; Glu --> Asp) of the RET proto-oncogene were recently reported to be found in 40% of sporadic medullary thyroid carcinomas (MTCs), the absence of missense mutations at codon 768 was confirmed both with PCR-restriction fragment length polymorphism (RFLP) and PCR-SSCP analysis in all examined cases of pheochromocytomas. These results suggest that mutations at codon 768 of the RET proto-oncogene do not represent a frequent mechanism of tumorigenesis for both sporadic and hereditary pheochromocytomas. PMID- 8732461 TI - Continuous stimulation of gonadotropin-releasing hormone (GnRH) receptors by GnRH agonist decreases pituitary GnRH receptor messenger ribonucleic acid concentration in immature female rats. AB - Although it is well recognized that continuous administration of gonadotropin releasing hormone agonist (GnRHa) induces pituitary desensitization, the precise molecular mechanism of this phenomenon is still unclear. To test the hypothesis that pituitary gonadotroph desensitization is mediated by a change in GnRH receptor (GnRH-R) gene expression, the GnRH-R mRNA concentration was analyzed in immature female rats during GnRHa treatment. Northern blot hybridization was used to determine the GnRH-R mRNA concentration several times after an injection of TAP-144-SR, a slow releasing GnRHa. The GnRH-R mRNA readings were 92.7 +/- 9.5%, 49.9 +/- 5.0%, 35.7 +/- 2.3% and 73.8 +/- 5.7% (Mean +/- SD) compared to each control value at 1, 2, 4 and 8 weeks, respectively, after a single injection of 0.94 mg TAP-144 SR. These changes in GnRH-R mRNA coincided with the changes in gonadotropin secretion and LH-beta mRNA in response to GnRH in the results of our previous report. The present results indicate that the reduction of the number of pituitary GnRH-R sites induced by continuous stimulation with GnRHa is regulated at a transcriptional level. PMID- 8732462 TI - Serum dehydroepiandrosterone (DHEA) and DHEA-sulfate (DHEA-S) in Alzheimer's disease and in cerebrovascular dementia. AB - A decreased concentration of dehydroepiandrosterone sulfate (DHEA-S) in patients with Alzheimer's disease (AD) has been reported but is still controversial. In the present study, serum concentrations of DHEA and DHEA-S were determined in 19 patients with AD, 21 patients with cerebrovascular dementia (CVD) and 45 age- and gender matched elderly control individuals from the Japanese community at large. Serum concentration of DHEA among controls, patients with AD and patients with CVD did not significantly differ from one another. However, patients with AD and patients with CVD were found to have lower concentration of serum DHEA-S and a lower DHEA-S/DHEA ration compared to normal control individuals. No significant difference was observed in the concentration of serum DHEA-S or the DHEA-S/DHEA ratio between patients with AD and those with CVD. These results suggest that reduced concentrations of serum DHEA-S may not be unique to AD, but instead reflect a common phenomenon in dementing diseases. However, since serum concentration of DHEA in these patients remained unchanged, the significance of DHEA in dementia remains unclear. PMID- 8732463 TI - Modified method using a somatostatin analogue, octreotide acetate (Sandostatin) to assess in vivo insulin sensitivity. AB - In order to evaluate the steady state plasma glucose (SSPG) method by using a new somatostatin derivative, octreotide acetate (Sandostatin) instead of somatostatin that we had used for the insulin sensitivity test, we examined whether octreotide was able to suppress C-peptide (CPR), glucagon (IRG), and GH to a similar degree to that achieved with somatostatin. A total of 52 studies were performed in 45 essential hypertensive subjects and 7 healthy subjects. Octreotide was given subcutaneously in a does of 50 micrograms or 100 micrograms 10 min before the test (sc 50, sc 100 groups) or intravenously infused over 2 h (10 micrograms in bolus followed by a constant infusion, 50, 100, or 150 micrograms/2 h: i.v. 50, i.v. 100, i.v. 150 groups). In all of the groups the plasma immunoreactive insulin (IRI) concentration increased gradually after insulin injection and reached the steady state plasma insulin (SSPI) level between 40 and 60 microU/ml at 60 min through 120 min. Plasma CPR at 120 min was the most suppressed (by 67% of the basal level in i.v. 150 group during the study period), but on the other hand in both the sc 100 and i.v. 100 groups the plasma CPR concentration at 120 min was suppressed by nearly 40%, but not significantly suppressed in either the sc 50 or the i.v. 50 group. Plasma IRG and GH were strongly suppressed after 60 min in all groups during the study period. Plasma glucose had increased significantly at 30 min and reached the steady state at 90 min through 120 min in hypertensive and healthy subjects. The results indicated that the modified SSPG method with continuous intravenous infusion of Octreotide at 150 micrograms/2 h was adequate for the measurement of insulin sensitivity. PMID- 8732465 TI - Management of hormone replacement therapy: the Swedish experience. AB - OBJECTIVE: To describe the experience of Swedish physicians in the management of hormone replacement therapy (HRT). METHODS: A postal questionnaire was sent to a sample of Swedish women aged between 46 and 62 years of age. The sample represented one woman in two from the birth cohorts 1946, 1942, 1938, 1934 and 1930. RESULTS: A total of 5990 women were sent the questionnaire and 4525 (76%) completed and returned it. Of this sample, 21% currently used HRT and a further 20% had used it in the past. A high proportion who continued using HRT derived benefit from it which was reflected in relief from vasomotor symptoms, irritability, insomnia, vaginal dryness and muscle and joint pain. Only 27% of those trying other, non-hormonal therapy considered that they had benefited from it. The major reasons for not taking HRT was fear of side-effects, a belief that the menopause should not be interfered with, and at the recommendation of their physician. The same reasons were given by those women starting HRT but discontinuing it later. CONCLUSIONS: The figure of 21% using HRT compares favourably with other Scandinavian countries. A high proportion of HRT is prescribed by gynaecologists reflecting that many GPs were unfamiliar with its use. When HRT is given by GPs, many favour the transdermal route. Compliance is a major problem and few women continue long enough to benefit fully. One way of minimizing this is to ensure that patients are fully informed and have any fears and misconceptions dealt with. Adverse and misleading information from the media may have to be countered. Individualization of therapy is important and should be tailored to particular age groups. Transdermal HRT may be more acceptable in some cases. PMID- 8732466 TI - The climacteric in The Netherlands: a review of Dutch studies on epidemiology, attitudes and use of hormone replacement therapy. AB - The climacteric has been the subject of research in the Netherlands for several decades. The incidence of climacteric symptoms has not changed since the pioneerwork of Jaszmann in 1967; nevertheless the number of contacts with a physician decreased by a factor of three during the 1970s and early 1980s. A rise in the number of contacts has occurred since 1987-1988 and nowadays the same number is reached as was 20 years ago. In 1970 psychopharmacons were the first choice of therapy. Nowadays 90% of the prescriptions for climacteric complaints are hormones. Twelve percent of women aged 45-60 years are current users of HRT. The mean duration of HRT use is only 7 months. The positive attitude of most women towards the climacteric is an explanation for this very short duration of use. Women receive their information on HRT use mainly through women's magazines and TV programmes and not through physicians. The determinants of longer duration of use are a positive attitude to treatment, younger age and the support of the GP. Physicians claim that they prescribe progestogens for 51% of oestrogen users, but the data give a figure of only 20% of prescription of progestogens with oestrogens. One explanation for this difference is the practice of many doctors to prescribe progestogens only once in 3 months. PMID- 8732467 TI - Management of hormone replacement therapy: the Italian experience. AB - OBJECTIVES: To review the characteristics of users and prescribers of hormone replacement therapy (HRT) in Italy, to attempt to explain the low uptake of this therapy. METHODS: A review of the published literature and prescription data. RESULTS: As in many other Western countries, gynaecologists are the main prescribers of HRT in terms of the number of prescriptions/physician/year. However, because general practitioners renew specialists' prescriptions several times a year, they are responsible for the greatest prescription volume. Overall GPs are responsible for 77.6% of HRT prescriptions while gynaecologists account for 20.7%. The attitude of gynaecologists and GPs to HRT is still equivocal and this may be a major factor in the low uptake. The percentage uptake is higher in northern and central Italy than in the south and it has been suggested that women in the hotter southern areas may suffer less from hot flushes. The evidence for this, however, is anecdotal. A recent study in Milan showed that the use of HRT was strongly correlated to social class and level of education. Women with 12 years of education or more were three times more likely to use HRT than those with less than 7 years education. There seemed to be no relationship between marital status and HRT use but fewer parous women used it and use decreased with an increasing number of children. In addition postmenopausal osteoporosis is not yet a registered indication for transdermal HRT, the major route of HRT administration in Italy. CONCLUSIONS: An increased uptake is expected in the future due to increased media interest and the greater availability and steady increase of transdermal HRT use over the last 5 years. PMID- 8732468 TI - HRT management: the American experience. AB - OBJECTIVES: To review the history, current clinical practice, choice of methods and number of prescriptions and sales of hormone replacement therapy (HRT) in the US. METHODS: Literature review, survey data collection, sales and prescription data. RESULTS: The number of women currently utilizing HRT is greater in women aged 40-60 (35%) but falls with advancing age greater than 65 (15%) and declines further in women greater than 80 (7%). News media and physicians represent the largest source of information used by US women for information regarding HRT. Obstetricians and gynecologists currently are the predominant physicians prescribing HRT, but in the future, family practitioners and paramedical workers will prescribe HRT. Women who spontaneously develop menopause early and younger women undergoing castration are more likely to take transdermal estrogen. However, the majority of prescriptions in the US are oral estrogens (86% of the market). The market leader of oral estrogens in the US are conjugated equine estrogens (70%). Fifty percent of prescriptions in women with a uterus consist of combined continuous estrogen and progestogen whereas as 42% consist of cyclic estrogen and progestogen. CONCLUSIONS: It appears that although there is an increase in HRT use by postmenopausal women in the US, the actual percentage of current users remains lower than anticipated. This occurred in spite of widespread media and educational efforts of benefits of HRT. Many women still fear the risk of breast and uterine cancer as well as the side effects of estrogen (primarily bleeding) which reduces the number of women currently taking estrogen. A number of surveys suggested that most women receive their information from the media and an equal amount from their physicians. With respect to estrogen use alone the majority of prescriptions are written for women without a uterus. Transdermal estrogens are used more commonly in women in the early postmenopausal period. In women with a uterus, most US physicians prescribe a combination of estrogen plus a progestogen, but when they do they utilize oral estrogen rather than transdermal estrogen. PMID- 8732469 TI - What women want from hormone replacement therapy: results of an international survey. AB - OBJECTIVES: To find out what women in France, Germany and the UK want from hormone replacement therapy (HRT). METHODS: A questionnaire sent to almost 2000 women in France, Germany and the UK followed up by more detailed interviews with 100 women from each country. RESULTS: From the questionnaire sent to 1871 women aged 48-55 years in France, Germany and the UK, it was found that 72% had not used HRT. The main reason for this seemed to be the prescribing physician who did not mention or recommend HRT. Some cultural differences were apparent. For example, in the UK 10% of the women did not use HRT because of perceived contraindications while the corresponding figures in France and Germany were 20% and 23%, respectively. Fear of side-effects was most common in Germany (20%), followed by the UK (14%) and much less in France (8%). In the detailed interviews with 100 women from each country, based on their menopausal status and the fact that they had received HRT, it was found that the main motivating factor was a desire to avoid the effects of ageing and prevent osteoporosis. In spite of their expectations being largely met, about one-third of the women sampled stopped taking HRT, mainly due to a dislike of breakthrough bleeding. CONCLUSIONS: The women who took HRT generally found it a positive experience and women who did use it derived considerable benefit. However, the main determinant whether HRT was used or not was the attitude of the patient's physician. PMID- 8732470 TI - Handling hormone replacement therapy: key issues for the prescriber. AB - OBJECTIVES: To examine the key considerations for physicians when prescribing hormone replacement therapy (HRT). METHODS: A review of the literature, including some unpublished data. RESULTS: As far as oral HRT is concerned, relatively large doses may have to be given because of the first-pass effect involving metabolism by the liver following absorption from the G1 tract. Alternatives include percutaneous gels, impregnated pessaries, subcutaneous implants and transdermal patches. The last of these offers the possibility of delivering oestrogen accurately in convenient form and many patients regard this as a more 'natural' way of treating the menopause. Provided equivalent doses of hormone are given, all these routes of administration are equally effective in preventing osteoporosis. The situation with respect to cardiovascular disease is more complex and different types of oestrogen or progestogen and different routes of administration can have different effects on metabolic risk factors. Certain oral HRT can adversely affect glucose tolerance and insulin metabolism while transdermal HRT has very little effect. CONCLUSIONS: The first issue for the physician is the choice of route of administration. To some extent this may depend on the patient's other clinical conditions such as raised triglycerides HRT. Patient preference is very important since this will affect compliance. One approach is to start with a fixed combination of oestrogen and progestogen which is usually easier for the patient to remember and use. Side-effects are usually due to the progestogen component. The variety of progestogens available in Europe means that switching to other combinations is not difficult if side effects occur. Patients satisfaction is the ultimate determinant. PMID- 8732471 TI - Menorest: technical development and pharmacokinetic profile. AB - Transdermal application of oestradiol enables the use of lower doses than with the oral drug and avoids first-pass metabolism in the liver. First-generation transdermal delivery systems were of reservoir design. Menorest is one of a new generation of patches, in which oestradiol is dispersed in a micronized suspension throughout the adhesive matrix. This design results in a very thin patch with good cosmetic acceptability. Pharmacokinetic studies indicate that Menorest allows transdermal release of oestradiol at a constant and reproducible rate at doses from 25-100 micrograms/day. There is a linear relationship between the dose of oestradiol administered (which is determined by the surface area of the patch) and the plasma concentration of oestradiol. In contrast to the standard reference reservoir patch, Estraderm, Menorest maintains plasma oestradiol concentrations at or above the target level of 40 pg/ml throughout the 84-h dosing interval. PMID- 8732472 TI - Efficacy and safety of Menorest in two positive-controlled studies. AB - OBJECTIVES: To evaluate the safety and efficacy of Menorest 50 in two clinical trials. METHODS: Menorest 50 was evaluated in two trials: the first was a parallel-group, randomized double-bind study vs. Premarin 0.625 mg/day in 214 women while the second was a parallel-group, open-label trial versus Estraderm TTS 50 in 205 women. RESULTS: In both studies there was a significant decrease in the mean number of hot flushes/day compared with baseline but no significant differences between treatments. The severity of hot flushes and the incidence and severity of other menopausal symptoms such as sweats, palpitations, headaches, depression, tiredness, vaginitis, loss of libido and dyspareunia were reduced to the same extent by all treatments. There were no significant differences between the different treatments as far as serum oestradiol or the incidence of systemic adverse events were concerned. In addition, small positive effects of total cholesterol and high-density lipoproteins were observed in the first study. Only a small reduction in cholesterol was seen in the second study in both groups but Menorest appeared to be better tolerated and a lower incidence of erythema, pruritus and other topic adverse events was reported. In addition, the twice weekly application of Menorest was found to be convenient. CONCLUSIONS: Menorest appears to be as equally effective as oral and transdermal oestradiol as far as reduction in the incidence and severity of menopausal symptoms is concerned. It was safe and also better tolerated than Estraderm. PMID- 8732473 TI - Osteoporosis prevention clinical study program. AB - OBJECTIVES: Oestrogens are widely believed to be effective against postmenopausal osteoporosis. However there are some outstanding questions which still need an answer. For example, the minimal effective dose regimen of oestradiol needs to be established and the relationship between oestradiol levels and efficacy on bone turnover and bone mass needs to be further clarified. METHODS: Menorest is being tested in the prevention of postmenopausal bone loss. A phase II/III clinical program, that includes two double blind, dose-ranging, placebo-controlled, parallel group, 2-year studies, has started in 58 centers in Europe and South Africa. Four-hundred eighty women will be enrolled in the two studies (201 and 305). The objective of the studies is to evaluate the efficacy of Menorest at different doses and regimens, in the prevention of bone loss in early postmenopausal women. In study 201, the treatment regimen is 'cyclic sequential' (24 days of transdermal oestradiol during a 28-day cycle with progestin taken during the last 14 days of oestrogen administration). In study 305 the treatment regimen is "continuous sequential' (28 days of transdermal oestradiol during, a 28-day cycle with progestin taken during the last 14 days of oestrogen administration). The doses studied are 50, 75, 100 micrograms/day in study 201, and 25, 50, 75 micrograms/day in study 305, (the two studies are otherwise identical). All 'active-dose' treated groups receive dydrogesterone 20 mg/day during the last 14 days of Menorest administration and placebo tablets are given to the placebo patch group. The main entry criteria are natural or surgical menopause, (with hormonal confirmation) from 1 to 6 years, with no contra indication to HRT and with a bone mineral density (BMD) at the lumbar spine with a T-score between 0 and -3. Women with severe vasomotor symptoms are excluded from the studies. The primary efficacy variable is the mean change from baseline, measured with dual energy X-ray absorptiometry (DXA) at 2 years, in the lumbar spine BMD (L1-L4). Whole body and hip BMD are also evaluated. Markers of bone turnover (bone-specific alkaline phosphatase, osteocalcin and CrossLaps) are monitored throughout the study. Blood samples are drawn on the third day of patch application at certain visits in order to monitor oestradiol levels and establish any potential correlation with activity on bone (BMD, bone markers). Besides routine safety analysis, lipid profile and coagulation factors are also monitored. Special attention is drawn to endometrial safety with endometrial aspiration or trans vaginal sonography (TVS) performed before study start, after 1 year and at 2 years of treatment. RESULTS: Data presented here refer to 146 patients for whom demographics and clinical data are already available, and to 370 patients for whom baseline DXA data have already been validated. The mean (+/ S.D.) age of the women included in the two studies is 53.4 (+/-3.2) with a menopausal age of 38.3 (+/-19.6) months. None of the women who entered the study had severe postmenopausal symptoms as shown by a mean number of hot flushes of 2.2 (+/-2.6) per day, during the last 14 days before inclusion. The mean (+/ S.D.) lumbar spine (L1-L4) BMD is 0.914 (+/-0.122) g/cm2 which corresponds to a Z score of -0.26 and a T-score of -1.17. Femoral neck, trochanter and Wards triangle have a BMD which is below the mean of age-matched controls but still within the normal range (Z-scores between 0 and -1). Only the whole body BMD is over the mean of age-matched controls, with a Z-score of 0.32. The in-vivo precision mean (+/-S.D.), was calculated and showed a value of 0.868 (+/-0.872), which can be considered a good performance. CONCLUSIONS: In summary, the use of one of the most recent techniques to assess the bone mineral content/density together with an accurate quality control program on all the densitometers used in the studies will help to improve the in-vivo BMD precision and therefore mak PMID- 8732474 TI - HLA-DQA1 and -DQB1 genotyping by PCR-RFLP, heteroduplex and homoduplex analysis. AB - PCR-RFLP typing methods for DQA1 and DQB1 in conjunction with the analysis of heteroduplex and homoduplex patterns have allowed a simple method for typing all of the major DQA1 and DQB1 alleles. This method has advantages over PCR amplification with sequence-specific primers (PCR-SSP), PCR hybridization with sequence-specific oligonucleotide probes (PCR-SSO) and other PCR-RFLP strategies for typing DQ alleles. The analysis of heteroduplex and homoduplex patterns can be used in conjunction with other PCR typing systems such as PCR-SSP as a confirmatory step with little additional work. In addition, a PCR-RFLP strategy was designed for resolving the DQB1*0602 and DQB1*0603 alleles, which involved the use of a primer containing a base mutation, creating a new restriction site which distinguished the two alleles. These techniques have enabled resolution of the major homozygous and heterozygous combinations of these DQA1 and DQB1 alleles. The PCR-RFLP technique does not require the large number of oligonucleotides that are necessary for both the PCR-SSP and PCR-SSO techniques and is thus both time and cost effective for infrequent or small numbers of samples. PMID- 8732475 TI - Developmental expression of the mouse MHC Q genes. AB - To investigate the role of class Ib genes in the Q region of the mouse major histocompatibility complex (MHC), the developmental expression and foetal tissue distribution of the Q genes were studied in the C57BL/6 mouse. Using RNase protection assays, we examined Q gene expression in a wide spectrum of foetal tissues at different stages of gestation, Q4, Q6 and Q8 were widely expressed in a variety of tissues. In contrast, the expression of Q1 was restricted to the thymus and intestinal epithelium. PMID- 8732476 TI - An apparent functional correlation between variations in amino acid residues in HLA-DR4.1 and 4.2 serological subtypes and oligonucleotide characterization. AB - HLA-DR4 can be subdivided serologically into two specificities, DR4.1 and DR4.2, using well-defined monospecific alloantisera used in the 11th International Histocompatibility Workshop. In this study, a total of 1095 random DR4-positive individuals from several ethnic groups were tested first for serotype DR4.1/4.2 and then for DRB1*04 alleles using polymerase chain reaction (PCR) followed by sequence-specific oligonucleotide probe hybridization (SSOPH). An almost 100% correlation between samples testing positive for DR4.1 and the presence of alanine at position 74 was observed, while samples testing positive for DR4.2 correlated with the presence of glutamic acid at position 74. DRB1*04 alleles 0401, 0402, 0404, 0405, 0408, 0409 and 0410 are aligned in functional groups which coincide with the serological subtype of DR4.1. DRB1*04 alleles 0403, 0406, 0407 and 0411 coincide with subtype DR4.2. Amino acid substitutions at positions 57, 71 and 86 indicate other significant variations between alleles within the serological subgroup of DR4.1 and define five minor subgroups. The serologic and oligonucleotide allelic subgroups are in turn correlated with recognized cellular Dw antigens. While sequence data provide evidence of structural differences, data on cellular antigens support a functional association between these designated groups and their significance in transplantation and GVHD. Testing results are categorized by ethnic group in order to establish frequency data for donor selection criteria. PMID- 8732477 TI - Investigation of the association of major histocompatibility complex genes, including HLA class I, class II and TAP genes, with clinical forms of Crohn's disease. AB - The aim of this study was to determine immunogenetic markers of susceptibility in Crohn's disease (CD), taking the different features of the clinical course of the disease into account. HLA class I, HLA class II and TAP transporter gene polymorphisms were studied using DNA typing methods. Gene and antigen frequencies were analysed and compared in a group of 102 CD patients and 200 unrelated healthy controls from the same area. Analysis of the whole CD patient population revealed no definite association with either HLA or TAP gene alleles, with the exception of an association with DRB1*1302 (Pc < 0.05). However, when clinical subgroups of patients were considered, specific associations with some genetic markers were found. The most definitive results involved a genetic association in the group of patients who did not respond to glucocorticoid therapy. This group was characterized by a high frequency of HLA-DRB1*04 (P < 0.05). Conversely, a positive association with the TAP2-A allele was found in cortico-responder patients (Pc < 0.03). Furthermore, analysis of the distribution of HLA class II alleles in relation to the presence of extra-intestinal manifestations revealed an association with the DQB1*0501 or *0503 suballele of DQ5 (P < 0.05). Finally, patients with lesions in the small bowel were more frequently HLA DRB1*07 (P < 0.05). The present study supports the concept of clinical heterogeneity in Crohn's disease associated with a background of genetic heterogeneity. PMID- 8732478 TI - HLA-G polymorphism and allelic association with HLA-A in a Finnish population. AB - The polymorphism of the HLA-G gene can be identified by PCR-RFLP analysis. This short communication describes the PCR-RFLP analysis of HLA-G polymorphisms in exons 2 and 3 and the association of different HLA-G and HLA-A alleles in 26 healthy Finnish families. PMID- 8732479 TI - Nomenclature for factors of the HLA system, update November 1995. PMID- 8732480 TI - TCR gene polymorphisms and autoimmune disease. AB - Autoimmunity may result from abnormal regulation within the immune system. As the T cell is the principal regulator of the immune system and its normal function depends on immune recognition or self/non-self discrimination, abnormalities of the idiotypic T-cell receptor (TCR) may be one cause of autoimmune disease. The TCR is a clonally distributed, cell-surface heterodimer which binds peptide antigen when complexed with HLA molecules. In order to recognize the variety of antigens it may possibly encounter, the TCR, by necessity, is a diverse structure. As with immunoglobulin, it is the variable domain of the TCR which interacts with antigen and exhibits the greatest amount of amino acid variability. The underlying genetic basis for this structural diversity is similar to that described for immunoglobulin, with TCR diversity relying on the somatic recombination, in a randomly imprecise manner, of smaller gene segments to form a functional gene. There are a large number of gene segments to choose from (particularly the TCRAV, TCRAJ and TCRBV gene segments) and some of these also exhibit allelic variation. Finally, polymorphisms in non-coding regions of TCR genes, leading to biased recombination or expression, are also beginning to be recognized. All these factors contribute to the polymorphic nature of the TCR, in terms of both structure and repertoire formation. It follows that inherited abnormalities in either coding or regulatory regions of TCR genes may predispose to aberrant T-cell function and autoimmune disease. This review will outline the genomic organization of the TCR genes, the genetic mechanisms responsible for the generation of diversity, and the results of investigations into the association between germline polymorphisms and autoimmune disease. PMID- 8732481 TI - Steroid hormones and adipose tissue. PMID- 8732482 TI - Identification of oestrogen receptors and oestrogen receptor mRNA in human adipose tissue. AB - The distribution of adipose tissue has a major impact on the morbidity and mortality associated with obesity. Furthermore, the distribution of adipose tissue seems to be regulated by sex hormones. Controversy exists over whether the effects of sex hormones (oestrogen and testosterone) on human adipose tissue are an indirect or a direct effect as contradictory results have been obtained when investigating the existence of these receptors in human adipose tissue. In the present study the authors reinvestigated the possible existence of oestrogen receptors (ERs) in human adipose tissue. Human adipocytes from both genders were found to contain specific oestrogen binding sites determined by ligand-binding techniques. The binding protein had a molecular weight of 65 kD (which is similar to that of the ER found elsewhere) and it was found that adipocytes contained mRNA encoding the ER. Moreover, human preadipocytes had no oestrogen-binding capacity and did not possess mRNA encoding the ER. Finally, the authors detected regional differences in receptor density. Women had an equal oestrogen-binding capacity in adipose tissue from the subcutaneous abdominal and the visceral depot, whereas men had twice as high oestrogen-binding capacity in subcutaneous adipose tissue compared with adipose tissue in the visceral fat depot. These findings indicate that mature human adipocytes possess ERs and thus, might be an oestrogen-responsive tissue and that oestrogen may be acting directly in mature adipocytes via its specific receptor. Human preadipocytes, however, seemed not to be an oestrogen-responsive tissue. Finally, preliminary data suggest that there might be differences in ER densities in different fat depots. PMID- 8732483 TI - Erythropoietic protoporphyria: two populations of reticulocytes, with and without protoporphyrin. AB - Erythrocytes from patients with erythropoietic protoporphyria contain large amounts of protoporphyrin. The photosensitivity experienced by these patients is assumed to be due to a leakage of protoporphyrin from the erythrocytes and transfer to the skin, where protoporphyrin acts as a photosensitizer. The leakage of protoporphyrin from the erythrocytes has been offered as an explanation for the great variety in protoporphyrin content observed among erythrocytes in this disease. Based on density gradient separation of red cells, it has been concluded that all reticulocytes and young erythrocytes contain large amounts of protoporphyrin. From our results, density gradient centrifugation is not suitable for age separation of red cells from patients with erythropoietic protoporphyria. By developing a new method for isolation of reticulocytes and applying flow cytometry to determine protoporphyrin content in individual cells, it was observed that two populations of reticulocytes were present in patients with erythropoietic protoporphyria, one with and the other without protoporphyrin. The half-life of protoporphyrin in red cells was found to be 12-14 days, in contrast to 1-2 days described previously, suggesting a slower release of protoporphyrin from the red cells than previously anticipated. PMID- 8732484 TI - Effect of regular physical training on age-associated alteration of body composition in men. AB - Body composition changes with increasing age in men, in that lean body mass decreases whereas fat mass increases. Whether this altered body composition is related to decreasing physical activity or to the known age-associated decrease in growth hormone secretion is uncertain. To address this question, three groups of healthy men (n = 14 in each group), matched for weight, height and body mass index, were investigated using dual-energy X-ray absorptiometry, indirect calorimetry and estimate of daily growth hormone secretion [i.e. plasma insulin like growth factor I (IGF-I-) levels]. Group 1 comprised young untrained subjects aged 31.0 +/- 2.1 years (mean +/- SEM) taking no regular physical exercise; group 2 consisted of old untrained men aged 68.6 +/- 1.2 years; and group 3 consisted of healthy old men aged 67.4 +/- 1.2 years undergoing regular physical training for more than 10 years with a training distance of at least 30 km per week. Subjects in group 3 had for the past three years taken part in the 'Grand Prix of Berne', a 16.5-km race run at a speed of 4.7 +/- 0.6 min km-1 (most recent race). Fat mass was more than 4 kg higher in old untrained men (P < 0.01, ANOVA) than in the other groups (young untrained men, 12.0 +/- 0.9 kg; old untrained men, 16.1 +/- 1.0 kg; old trained men, 11.0 +/- 0.8 kg), whereas body fat distribution (i.e. the ratio of upper to lower body fat mass) was similar between the three groups. The lean mass of old untrained men was more than 3.5 kg lower (P < 0.02, ANOVA) than in the other two groups (young untrained men, 56.4 +/- 1.0 kg; old untrained men, 52.4 +/- 1.0 kg; old trained men, 56.0 +/- 1.0 kg), mostly because of a loss of skeletal muscle mass in the arms and legs (young untrained men, 24.0 +/- 0.5 kg; old untrained men 20.8 +/- 0.5 kg; old trained men, 23.6 +/- 0.7 kg; P < 0.01, ANOVA). Resting metabolic rate per kilogram lean mass decreased with increasing age independently of physical activity (r = -0.42, P < 0.005). Fuel metabolism was determined by indirect calorimetry at rest. Protein oxidation was similar in the three groups. Old untrained men had higher (P < 0.001) carbohydrate oxidation (young untrained men, 13.2 +/- 1.0 kcal kg-1 lean mass; old untrained men, 15.2 +/- 1.3 kcal Kg-1; old trained men, 7.8 +/- 0.8 kcal kg 1), but lower (P < 0.05, ANOVA) fat oxidation (young untrained men, 10.1 +/- 1.2 kcal kg-1 lean mass; old untrained men, 6.5 +/- 1.0 kcal kg-1; old trained men, 13.7 +/- 1.0 kcal kg-1) than the other two groups. Mean plasma IGF-I level in old trained men was higher than in old untrained men (P < 0.05), but was still lower than that observed in young untrained men (P < 0.005) (young untrained men, 236 +/- 24 ng mL-1; old untrained men, 119 +/- 13 ng mL-1; old trained men, 166 +/- 14 ng mL-1). In summary, regular physical training in older men seems to prevent the changes in body composition and fuel metabolism normally associated with ageing. Whether regular physical training in formerly untrained old subjects would result in similar changes awaits further study. PMID- 8732485 TI - Synergistic stimulatory effects of tumour necrosis factor alpha and interferon gamma on replication of human immunodeficiency virus type 1 and on apoptosis of HIV-1-infected host cells. AB - Differential and sometimes contradictory effects have been described for tumour necrosis factor alpha (TNF-alpha) and interferon gamma (IFN-gamma) on replication of human immunodeficiency virus type 1 (HIV-1). The authors examined individual and coordinate action of these cytokines on HIV-1 expression, and on apoptosis of HIV-1-infected host cells by determination of reverse transcriptase activity in cell culture supernatant, expression of HIV-1-RNA and production of p24 antigen in the promonocytic cell line U937 and its persistently HIV-1-infected clone U1. Apoptosis was demonstrated by typical cleavage of cellular DNA at internucleosomal regions in promonocytic and T-lymphocytic cell lines. TNF-alpha alone markedly stimulated HIV-1 replication in U1 cells at the transcriptional and on the translational level. Exclusive application of IFN-gamma only slightly enhanced HIV-1 expression, whereas it synergistically potentiated stimulatory effects of TNF-alpha. Both cytokines also synergistically induced apoptosis in HIV-1-infected host cells. Co-ordinate action of TNF-alpha and IFN-gamma is suggested to represent an important mechanism for disease progression in HIV infection. These findings demonstrate that cytokine effects on viral expression may vary depending on their single or combined application. PMID- 8732486 TI - Echo-Doppler left ventricular filling abnormalities in patients with rheumatoid arthritis without clinically evident cardiovascular disease. AB - Our investigation aimed at verifying diastolic abnormalities in rheumatoid patients, without clinically evident cardiovascular disease and other confounding complaints, by using pulsed Doppler examination of transmitral blood flow. We selected 40 patients fulfilling revised American Rheumatism Association (ARA) criteria for the diagnosis of rheumatoid arthritis having no symptoms of cardiac disease or clinical findings of other extracardiac diseases. We also studied 40 rheumatoid-matched healthy volunteers as a control group. An echocardiographic examination was carried out on each subject. Left ventricular structural and functional measurements were obtained. Interventricular, septal thickness and left ventricular mass index were significantly higher in rheumatoid patients than in the control group. We also found in rheumatoid patients higher mean values of peak A velocity and A/E ratio. When multiple linear regression analysis was performed on the data of rheumatoid patients we found an independent relationship only between A/E ratio and left ventricular mass. In conclusion, our results confirm diastolic abnormalities in rheumatoid patients and point out that these abnormalities also affect echo-Doppler parameters of left ventricular filling. Moreover, further analysis of our data may suggest the possibility that structural left ventricle changes could be responsible for left ventricular filling impairment. PMID- 8732487 TI - Serum neurone-specific enolase as an indicator of stroke volume. AB - Serum neurone-specific enolase (NSE) and computerized tomography (CT) stroke volume were compared in patients admitted within 24 h of an acute stroke. Serum samples were obtained on admission and daily for the next 4 days. Of 163 patients, CT scans revealed 25 with intracerebral haemorrhages, one haemorrhagic infarct and 83 measurable acute infarcts. The serum NSE levels of those with infarcts was significantly higher than in those with haemorrhages at 48 (P = 0.0003) and 72 h (P = 0.04). The maximum serum NSE value tended to occur later in those with large infarcts (P = 0.0035). There was a significant correlation between infarct volume and serum NSE at 48 h (r = 0.27, P = 0.015) and 96 h (r = 0.27, P = 0.015) and with the maximum serum NSE over the 4 days (r = 0.36, P = 0.001). There was no significant correlation between haemorrhage volume and NSE. In conclusion, serum NSE may be a useful marker of infarct volume in studies of therapy in acute stroke. Sampling for NSE should continue, at least in those with large infarcts, for longer than 4 days. Serum NSE cannot be used to distinguish between haemorrhage and infarction in patients with an acute stroke. PMID- 8732488 TI - The effect of folic acid on the homocysteine metabolism in human umbilical vein endothelial cells (HUVECs). AB - Mild hyperhomocysteinaemia is associated with increased risk for vascular disease. We studied homocysteine export from human umbilical vein endothelial cells (HUVECs) by measuring total homocysteine (tHcy) concentrations in the culture medium. Under standard culture conditions tHcy concentrations in the HUVEC culture medium increased by constant amounts after 24, 48 and 72 h [mean = 2.5 (SD +/- 0.7) mumol L-1 homocysteine every 24 h]. As the cells are the only source of homocysteine increase in the culture medium, we designate this as homocysteine export from HUVEC. Folic acid supplementation to the culture medium lowered the homocysteine export in a dose-dependent manner. Methyl tetrahydrofolate (MeTHF) and folinic acid (a stable precursor of MeTHF) were in this respect about 10 times more effective than folic acid. A 50% reduction in the homocysteine export was seen with 10-30 nmol L-1 MeTHF supplementation; reduction to almost zero was seen with 100-300 nmol L-1 MeTHF. Additions to the culture medium of the other vitamins involved in the homocysteine metabolism, such as vitamin B12, vitamin B6 and flavin adenine dinucleotide, did not show any effect on homocysteine export. Because homocysteine export reflects an imbalance in the homocysteine metabolism, our observations showed a susceptible dependency of this metabolism on folic acid in endothelial cells. PMID- 8732489 TI - Differences in phospholipase A2 activity between males and females and Asian Indians and Caucasians. AB - There is epidemiological evidence that chronic inflammatory diseases occur more frequently in female than in male subjects and prevail differently in various ethnic populations. Phospholipase A2 (PLA2) (group II) plays a key role in many inflammatory reactions by releasing free arachidonic acid, which is a prerequisite for the production of proinflammatory lipid mediators. We therefore, measured PLA2 activity in plasma, serum, leucocytes and lymphocytes in 20 female and 20 male subjects, 10 of each group being of Asian Indian and of Caucasian origin respectively. When PLA2 activity was measured in crude plasma and serum no dependency from gender and ethnicity was observed. Following acid extraction and heating, PLA2 activity in plasma was higher in Caucasians (27.8 +/- 2.2 nmol L-1 mg-1 protein 60 min-1) than in Asian Indians (17.9 +/- 2.5 nmol L-1 mg-1 protein 60 min-1) (P < 0.005) and higher in females (28.5 +/- 2.6 nmol L-1 mg-1 protein 60 min-1) than in males (17.3 +/- 2.0 nmol L-1 mg-1 protein 60 min-1) (P < 0.001). Similar differences were observed when only Asian Indian or Caucasian females were compared with their corresponding males. Contrary to plasma, in which the specific activity of PLA2 increased following acid extraction and heating, the activity was completely abrogated in serum after extraction and heating. Lymphocytes exhibited lower activities of PLA2 than neutrophils in all four groups of subjects investigated. Females had a tendency towards higher PLA2 activity in both lymphocytes and neutrophils than males. In conclusion the present investigation revealed an ethnic and sex-dependent basal activity of PLA2, a key enzyme in the pathogenesis of chronic inflammatory diseases. PMID- 8732490 TI - Hyperlipoprotein(a)aemia in nephrotic syndrome. AB - The nephrotic syndrome is frequently associated with hyperlipidaemia and hyperfibrinogenaemia, leading to an increased coronary and thrombotic risk, which may be enhanced by high lipoprotein (a) [Lp(a)] concentrations. We followed the quantitative and qualitative pattern of plasma lipoproteins over 18 months in a patient with nephrotic syndrome suffering from premature coronary artery disease and with elevated level of Lp(a) (470 mg dL-1). Analysis of kinetic parameters after heparin-induced extracorporeal plasma apheresis revealed a reduced fractional catabolic rate for both low-density lipoprotein (LDL) and Lp(a). After improvement of the nephrotic syndrome, Lp(a) decreased to 169 mg dL-1 and LDL concentrations were normalized. The decrease of Lp(a) was associated with an increase in plasma albumin concentrations. Analysis of apo(a) isoforms in the patient showed the presence of isoform S2 (alleles 10 and 19). Consequently, the authors' present strategy is to normalize the elevated Lp(a) and fibrinogen levels. For this purpose heparin-mediated extracorporeal LDL precipitation (HELP) apheresis is a promising regimen, helping to reduce the thrombotic risk and prevent coronary and graft atherosclerosis as well as the progression of glomerulosclerosis in our patient. PMID- 8732491 TI - Quantitative and qualitative alterations of dystrophin are expressed in muscle cell cultures of Xp21 muscular dystrophy patients (Duchenne and Becker type). AB - The authors studied skeletal muscle cell cultures from four control subjects, two patients with Duchenne muscular dystrophy, two Duchenne carriers and three patients with Becker muscular dystrophy with different phenotypes. Western blotting was performed on well-differentiated myotubes and compared with the results obtained in muscle tissue. In all cultures the band of dystrophin closely corresponded to the one observed in muscle tissue: both quantitative and qualitative defects were observed. This confirms the early expression of the Xp21 gene defect in uninnervated muscle cultures and supports the usefulness of muscle cultures both in diagnostic procedure and as a model to study the disease. PMID- 8732492 TI - Arginine, lysine and ornithine as vasodilators in the forearm of man. AB - Nitric oxide is an endogenous vasodilator produced from L-arginine and oxygen by stereospecific enzymes. L-arginine itself can act as a vasodilator when administered at high doses to humans. This effect has been attributed by some to provision of extra substrate for production of nitric oxide. This work compares L arginine-induced vasodilation with that caused by D-arginine, hyperosmolar sodium chloride and by other cationic amino acids in the resting forearm vasculature of normal subjects. By these means we identify whether L-arginine-induced vasodilation has a component related to stereospecific provision of substrate for nitric oxide production, or whether it can be accounted for by other phenomena. Effects of hyperosmolar sodium chloride and both L and D isomers of arginine, lysine and ornithine on forearm blood flow in eight normal male subjects were compared by bilateral forearm venous occlusion plethysmography. Vasodilator responses to saline and each amino acid were compared as the area under dose response curves with single-factor analysis of variance (ANOVA). The magnitude of vasodilation obtained with the D isomers of each amino acid was compared with the L counterpart by application of single-factor ANOVA to the appropriate areas under dose-response curves. All three cationic amino acids increased forearm blood flow. Part of the increase could be related to the high osmolality of infusates-comparison with equiosmolar sodium chloride solutions shows that ornithine does not differ significantly as a vasodilator (P > 0.4), but that arginine and lysine have greater vasodilator effects than can be accounted for by osmolality alone (P < 0.001). The D isomers of arginine and lysine were more potent dilators than their L counterparts (arginine P < 0.03, lysine P < 0.02). The vasodilator effects of arginine in the forearm vascular bed at rest are not stereospecific, they are common to other cationic amino acids, greater for D isomers and occur only when normal plasma concentrations are raised far above the physiological range. These features suggest that the vasodilator effect of arginine is in part physical and related to the presence of the molecule in the vessel lumen. They do not suggest that increased provision of substrate, with a consequence of increased nitric oxide production, is the principal basis of L arginine-induced vasodilation in normal humans. PMID- 8732493 TI - Acute hyperinsulinaemia decreases cholesterol synthesis less in subjects with non insulin-dependent diabetes mellitus than in non-diabetic subjects. AB - To investigate the effect of insulin on cholesterol synthesis in vivo we measured plasma mevalonic acid (MVA) concentrations using gas chromatography-mass spectrometry in six non-obese patients with non-insulin-dependent diabetes mellitus (NIDDM) [four men, two women; age 57.5 +/- 2.2 years (mean +/- SEM); glycated haemoglobin (HbA1) 8.5 +/- 0.5%; total cholesterol (TC) 5.7 +/- 0.5 mmol L-1, triglyceride (TG) 3.8 +/- 0.9 mmol L-1] and six non-diabetic, sex- and age matched control subjects (age 55.7 +/- 2.8 years; HbA1 6.5 +/- 0.1%; TC 5.4 +/- 0.3 mmol L-1, TG 1.2 +/- 0.1 mmol L-1). Subjects were studied twice: during 13-h hyperinsulinaemic (1 mu kg-1 min-1), euglycaemic (5 mmol L-1) clamp and during a saline infusion. Baseline MVA concentration was significantly higher in diabetic patients than in control subjects (9.8 +/- 0.7 ng mL-1 vs. 5.6 +/- 0.9 ng mL-1, P = 0.004). At the end of each study, MVA concentration, expressed as a percentage of baseline, was significantly lower during the hyperinsulinaemic, euglycaemic clamp than during the saline study in both the diabetic (54.4 +/- 5.3% vs. 69.6 +/- 6.3%, P = 0.036) and control subjects (30.5 +/- 3.4% vs. 61.7 +/- 6.0%, P = 0.01). However, the decrease in MVA during the hyperinsulinaemic clamp study was more marked in the control subjects than in the diabetic subjects (P = 0.03). A significant positive correlation was found between percentage decrease of MVA and non-esterified fatty acids following the insulin clamp in NIDDM (r = 0.83, P = 0.04). We conclude that acute hyperinsulinaemia decreases cholesterol synthesis less in subjects with NIDDM than in non-diabetic subjects and that this phenomenon, together with increased basal cholesterol synthesis in NIDDM, may in part be due to insulin resistance. PMID- 8732494 TI - Nisoldipine attenuates myocardial stunning induced by multiple coronary occlusions in conscious pigs and this effect is independent of changes in hemodynamics or coronary blood flow. AB - Recent studies suggest that calcium channel blockers attenuate reversible post ischemic myocardial dysfunction (myocardial "stunning") in vivo. This beneficial effect, however, has been shown either in open-chest preparations, which are subject to the confounding influence of many unphysiological conditions, or in models in which treatment caused significant hemodynamic alterations. Furthermore, all of the studies have been conducted in the dog, and almost all of them have examined the effect of calcium antagonists after a single ischemic episode. The goal of the present investigation was to assess the effect of nisoldipine in a conscious pig model of repetitive ischemia, and to determine whether the drug exerts direct cardioprotection independent of hemodynamic changes. A total of 33 conscious pigs were used. Pigs underwent a sequence of 10 2-min coronary occlusions, each separated by 2 min of reperfusion, and were randomly assigned to a treated group (n = 11), in which nisoldipine was infused at a rate of 0.5 microgram/kg/min from 15 min before the first coronary occlusion till 30 min after the last reperfusion, and a control group (n = 12), which received vehicle. Results showed that there were no significant differences between the two groups with respect to ischemic bed size or hemodynamic variables throughout the experiment. Collateral blood flow to the ischemic regions was virtually nil in both groups. During the sequence of coronary occlusions, systolic thickening fraction in the ischemic region decreased similarly in the two groups. After the 10th reperfusion, however, the recovery of wall thickening was markedly enhanced in treated compared to control pigs, with the differences being statistically significant at 5, 15, and 30 min and 1, 3, 4 and 5 h. The total deficit of wall thickening after the 10th reperfusion (an integrative assessment of post-ischemic dysfunction) was 51% less in the treated compared with the control group (P < 0.001). This study demonstrates that nisoldipine markedly attenuates myocardial stunning after multiple ischemic episodes in conscious pigs, the improvement is evident immediately after the end of the ischemic episodes and is sustained throughout the recovery phase. This beneficial effect is independent of any favourable hemodynamic changes, and therefore indicates a direct cardioprotective action of nisoldipine. PMID- 8732495 TI - Analysis of the ionic basis for cocaine's biphasic effect on action potential duration in guinea-pig ventricular myocytes. AB - The effects of cocaine on the duration of the cardiac action potential were investigated in isolated guinea-pig ventricular myocytes at 37 degrees C. Following a 10-min exposure of cells to 3 microM cocaine, APD90 increased significantly by +22 +/- 5% (n = 6). In contrast, following a ten minute exposure to 30 or 100 microM cocaine, APD90 was reduced by -24 +/- 6% (n = 5) and -53 +/- 2% (n = 8), respectively. The ionic basis for cocaine's effects on the APD was investigated using the whole cell voltage-clamp technique at 37 degrees C. Cocaine produced a concentration-dependent reduction in the amplitude of IK tail currents with an estimated IC50 of 4 microM. The kinetics and voltage dependence of the cocaine-sensitive current indicate that cocaine selectively blocks a current identical to the E-4031 sensitive current IKr. No significant reduction of the slow component of IK (IKs) was observed during exposure to 30 or 100 microM cocaine. High (30 and 100 microM) concentrations of cocaine also produced a significant reduction of both the L-type calcium current and the TTX-sensitive plateau current. Pre-treatment of cells with 10 microM TTX also converted the APD shortening effect of 30 microM cocaine to one of APD-prolonging. This implies that cocaine block of a TTX-sensitive window current contributes to the APD shortening effects produced by high concentrations of cocaine. We conclude that: (1) cocaine produces a biphasic concentration-dependent effect on repolarization in guinea-pig ventricular myocytes; and (2) this biphasic effect on repolarization results from differences in the sensitivity of inward and outward currents to the blocking effects of cocaine. PMID- 8732496 TI - Ventricular arrhythmias induced by ischaemia-reperfusion are unaffected by myocardial glutathione depletion. AB - Reduced glutathione (GSH) is a major myocardial antioxidant. Since reperfusion phenomena such as ventricular fibrillation (VF) are associated with oxygen free radical production during ischaemia, myocardial GSH depletion might be expected to increase susceptibility to such phenomena. This possibility was tested in isolated rat hearts using diethylmaleate (DEM) or L-buthionine-SR-sulfoximine (BSO) to deplete myocardial GSH. High dose DEM (860 mg/kg) depleted myocardial GSH from a control mean of 7.64 +/- 0.73 to 3.18 +/- 0.56, low dose DEM (215 mg/kg) to 4.29 +/- 0.53 nmol/mg protein and BSO (4 mmol/kg) from a control mean of 6.94 +/- 0.54 to 2.18 +/- 0.14 nmol/mg protein. Hearts were perfused in the Langendorff mode at 37 degrees C with bicarbonate buffer (K+ = 4.3 mM). Regional ischaemia was induced for 5, 8.5, 10, 20 or 40 min (DEM groups: n = 10/treatment/time point) or 8.5 min only (BSO groups: n = 10/treatment) then hearts were reperfused for 5 min. Reperfusion VF incidence showed a classical "bell-shaped" curve, but there was no difference in VF incidence, VF time-to onset, arrhythmia duration and "arrhythmia scores" between GSH-depleted and control hearts. Depleting myocardial GSH is not proarrhythmic for reperfusion induced arrhythmias. It would appear GSH is not significantly involved in protecting against the oxidant stress of reperfusion, or conversely that the reserve of this redox system is so high only severe depletion might show an effect. PMID- 8732497 TI - Onset of reduced Ca2+ sensitivity of tension during stunning in porcine myocardium. AB - Recent data suggest that reduced Ca2+ sensitivity of tension is a mechanism of the post-ischemic myocardial dysfunction, termed stunning. The purpose of the present study was to determine whether the decrease in myofilament Ca2+ sensitivity occurs during ischemia or during the subsequent period of reperfusion. Serial biopsies from an in vivo open-chest porcine model of regional LAD myocardial stunning (n = 6) were used to obtain in vitro measurements of Ca2+ sensitivity of tension in myocardium from the LAD bed. Regional ventricular myocardial function was assessed from percentage systolic myocardial wall thickening (%Th) and the load-independent end-systolic pressure wall thickness relation (ESPTR). Stunning was induced by 45 min of low flow LAD ischemia (43 +/- 4 ml/min/100 gm) followed by 30 min of reperfusion with control aerobic flow (117 +/- 7 ml/min/100 g). Endocardial biopsies were obtained from the LAD bed during pre-ischemia, ischemia (immediately prior to reperfusion), and post-ischemia (after 30 min of reperfusion). Biopsies were mechanically disrupted and single cell-sized preparations of permeabilized myocardium were attached to a force transducer to measure directly steady-state tension-pCa relationships. The % decreased to 7 +/- 11% of control during ischemia (P < 0.001) and returned to 30 +/- 11% of control in the post-ischemic stunned state (P < 0.001). Stunning resulted in a significant leftward shift of the ESPTR as compared to control, indicating depressed regional myocardial function. The pCa (-log[Ca2+]) for half maximal activation of tension, i.e. pCa50, was 5.96 +/- 0.04 in control myocardium and was unchanged during ischemia (5.95 +/- 0.03), but significantly decreased to 5.82 +/- 0.04 upon reperfusion (P < 0.05). These data show that the decrease in Ca2+ sensitivity of tension associated with stunning occurs during reperfusion, and supports the idea that reperfusion injury is a mechanism of myocardial stunning. PMID- 8732498 TI - Extracellular matrix collagen synthesis and degradation following coronary balloon angioplasty. AB - Percutaneous transluminal coronary angioplasty is associated with intimal hyperplasia and extracellular matrix deposition of collagen, leading to restenosis in a significant number of cases. The purpose of the present study was to determine the effects of balloon angioplasty on extracellular matrix collagen content and collagenase activity in a porcine coronary artery restenosis model 6 weeks following balloon injury. We tested the hypothesis that in balloon-injured arteries the neointimal extracellular matrix was characterized by increased collagen content and decreased metalloproteinase activity relative to non-injured arteries. Male miniswine maintained on a high cholesterol diet underwent cardiac catheterization and double balloon injury to the right and left circumflex coronary arteries. The coronary arteries were either pressure-perfusion-fixed and prepared for histological examination, or dissected free of adventitia for further collagen and matrix metalloproteinase studies. Collagen synthesis in balloon-injured coronary arteries was compared to non-injured arteries using Northern blot analysis and histochemical stains. Comparative studies on differences between balloon-injured and non-balloon-injured arterial matrix metalloproteinase activity were done using zymography. Balloon angioplasty arterial injury resulted in a significant increase in type I collagen mRNA expression, with increased collagen deposition in the extracellular matrix. In contrast, matrix metalloproteinase activity was markedly decreased. The results suggest that the increased neointimal extracellular matrix observed late in the injury response may be due to not only increased collagen synthesis, but also reduced degradation. The failure to achieve a balance between the synthesis and degradation of extracellular matrix collagen could serve as an important mechanism responsible for restenosis. PMID- 8732499 TI - Endothelin-1 and vasopressin activate Ca(2+)-permeable non-selective cation channels in aortic smooth muscle cells: mechanism of receptor-mediated Ca2+ influx. AB - The effects of vasopressin and endothelin-1 on cultured aortic smooth muscle cell lines (A7r5) were investigated by measurements of intracellular calcium [Ca2+]i and the patch-clamp techniques. Vasopressin and endothelin-1 (100 nM) evoked an initial peak followed by a smaller sustained rise of [Ca2+]i in the presence of extracellular calcium [Ca2+]o. In the absence of [Ca2+]o, only the initial peak of [Ca2+]i was observed. Therefore, the initial peak of [Ca2+]i was mainly due to calcium release from the storage sites, whereas the later sustained rise of [Ca2+]i was due to the calcium entry from outside. The sustained rise of [Ca2+]i was unaffected by nifedipine (10 microM) significantly, but was completely abolished by La3+ (1 mM). Under current clamp conditions with K(+)-internal solution, vasopressin and endothelin-1 (100 nM) produced hyperpolarization, then followed by depolarization. Under voltage clamp conditions at a holding potential of -40 mV, both vasopressin and endothelin-1 first activated the outward current, then followed by a long-lasting inward current with a high noise level. The first outward current was abolished by charybdotoxin (100 nM), Cs+ in the patch pipette and high EGTA (10 mM) in the pipette, suggesting that it was a Ca(2+)-sensitive K+ current (IK.Ca). The inward current was still elicited with the patch pipette containing Cs(+)-internal solution, and reversed at about 0 mV. The reversal potential was not significantly altered by the replacement of [Cl-]i or [Cl-]o, proposing that the inward current is a cation selective channel (IN.S.). The inward current was also observed even when extracellular cations are Ca2+. La3+ (1 mM), Cd2+ (1 mM) completely abolished the vasopressin-induced (IN.S.), however, nifedipine (10 microM) failed to inhibit it significantly. Single channel activities were recorded in the cell-attached configurations when vasopressin or endothelin-1 was applied to the bathing solution. The unitary conductance of the channels was approximately 20 pS with 140 mM Na+, Cs+, or K+ in the pipette, but was 15 pS with 110 mM Ca2+ in the pipette. Permeabilities sequence calculated from the reversal potentials was Na+ not equal to Cs+ not equal to K+ > Ca+. These results provide evidence that calcium entry and membrane depolarization elicited by vasopressin or endothelin-1 are mediated by a receptor mediated Ca(2+)-permeable non-selective cation channel in aortic smooth muscle cells. PMID- 8732500 TI - Contractile function of cardiomyocytes from the spontaneously hypertensive rat. AB - In essential hypertension it has been hypothesized that there is a generalized derangement in cellular Ca2+ homeostasis. The aim of this study was to assess whether there is functional evidence for increased vulnerability to Ca overload in cardiomyocytes derived from age-matched (11-week) normotensive Wistar-Kyoto (WKY) and spontaneously hypertensive rat (SHR) strains. Contraction cycles of isolated ventricular cardiomyocytes were recorded using a rapid digital imaging technique and were evaluated by computation of a range of normalized parameters. Significant differences in the basal contractile cycles of SHR and WKY ventricular cardiomyocytes recorded under control conditions (36 degrees C, 3 Hz, 1 mM Ca2+) were detected. SHR myocytes exhibited a reduction in maximum shortening attained (SHR/WKY = 0.79) and in maximum rates of shortening (SHR/WKY = 0.85) and lengthening (SHR/WKY = 0.87). In the SHR there was a small delay in excitation-contraction coupling latency and an abbreviation of the contractile cycle (SHR/WKY = 0.87). The time to peak contraction was not different in the two strains and was not altered by the inotropic interventions studied. No functional evidence for an increased susceptibility of the SHR myocytes to uncompensated "Ca2+ leakiness" was observed during repeated challenge of elevated extracellular Ca2+. In both strains, the relative increase in maximum shortening of myocytes to isoproterenol (10(-8) M) was similar, suggesting that differential sensitivity to adrenergic modulation is due to factors other than altered cardiomyocyte responsiveness. Diminished basal cardiomyocyte contractile function may represent an intrinsic feature of impaired cardiovascular function in this form of genetically determined hypertension. PMID- 8732501 TI - Adenovirus-mediated overexpression of human transforming growth factor-beta 1 in rat cardiac fibroblasts, myocytes and smooth muscle cells. AB - Transforming growth factor-beta 1 (TGF-beta 1) is known to regulate cardiac cell function and its overexpression in the heart is thought to contribute to the development of cardiac hypertrophy and fibrosis. We wished to develop a high efficiency gene transfer method that could be used both in vitro and in vivo and result in the overexpression of TGF-beta 1. For this purpose, we constructed a replication-deficient human adenovirus 5 vector encoding for human TGF-beta 1 and used for control purposes an adenovirus lacZ vector. The adenovirus 5 construct was capable of infecting neonatal rat cardiac myocytes, fibroblasts and VSMCs. Of the three cell types, cardiac myocytes appear more susceptible to infection by the adenovirus 5 construct as assessed through beta-galactosidase staining. Infection of cardiac fibroblasts, myocytes and VSMCs with the hTGF-beta 1 adenovirus leads to the expression of hTGF-beta 1 mRNA and enhanced levels of bioactive and total TGF-beta 1 protein. Infection with hTGF-beta 1 adenovirus also results in enhanced levels of collagen type III gene expression in VSMCs and fibroblasts whereas in cardiac myocytes it leads to increased levels for sarcomeric and beta-actin. Thus, this adenoviral vector might be used for the exploration of in vivo effects of altered levels of cardiac TGF-beta 1. PMID- 8732502 TI - Rapid onset of lysophosphatidylcholine-induced modification of whole cell cardiac sodium current kinetics. AB - Lysophosphatidylcholine (LPC), an ischemic metabolite implicated in arrhythmogenesis, has been shown to modulate aspects of Na+ channel gating, but its effects on steady-state availability (h infinity), recovery from inactivation, and the timing of onset and possible reversibility, have not been characterized. We studied Na current (INa) by the whole-cell patch clamp technique on isolated rat ventricular myocytes at 22 degrees C with reduced Na+ (45 mM out, 5 mM in) from a holding potential of -150 mV. Changes in the electrophysiological parameters were measured after LPC 10 microM was added to the bath and compared to time controls (TC) taken from the time of seal formation. LPC decreased peak current for a test potential to -30 mV by about 20%. The peak current voltage relationship shifted in a positive direction by about 5 mV after LPC as compared to a small 2 mV negative shift in TC cells. LPC shifted the steady-state availability curve in the hyperpolarizing direction by about 6 mV. LPC perfusion caused a slowing of the decay of INa, and also a slowing of recovery from inactivation. Onset of the effects occurred within 6 min after adding LPC to the bath and were statistically significant with respect to TC cells between 12 and 16 min. In three cells, some of the effects on INa were either arrested or partially reversed by washout and cell survival was less than 20 min if LPC was not removed from the bath. These LPC induced changes in INa would tend to slow conduction and increase refractoriness, effects also seen in acutely ischemic myocardium. We therefore conclude that LPC action on INa may potentiate the arrhythmogenic substrate and that the onset of these changes are sufficiently rapid to play a role in the electrical instability of acute ischemia. PMID- 8732503 TI - Depletion of energy reserve via the creatine kinase reaction during the evolution of heart failure in cardiomyopathic hamsters. AB - To study the contribution of myocardial energy reserve to the deterioration of cardiac function during the development of heart failure, we defined energy reserve via the creatine kinase (CK) reaction and the isovolumic contractile performance in hearts of cardiomyopathic hamsters at the ages of 1.5, 4, 17, 30 and 43 weeks and in age-matched normal hamsters. Energy reserve via the CK reaction was estimated by the product of total CK activity and the content of total creatine in the heart. Isovolumic contractile performance was measured as rate pressure product (RPP, 10(3) mmHg/min) in isolated hearts. Contractile reserve was assessed as the increase of RPP elicited by high calcium stimulation. Compared to the controls, decreases in total CK activity and content of total creatine were observed in hearts of 17-, 30- and 43-week-old cardiomyopathic hamsters. These changes were not observed in the skeletal muscle. Although the decrease of baseline RPP first occurred at the age of 30 weeks (11.5 +/- 0.9 v 20.5 +/- 0.8, P < 0.05), the contractile reserve was already reduced at the age of 17 weeks (9.9 +/- 1.3 v 23.6 +/- 1.9, P < 0.05). A linear relationship was found between the energy reserve via creatine kinase reaction and the contractile reserve of the heart (r2 = 0.85). Furthermore, concomitant decreases in the CK reaction velocity and the contractile reserve were observed in cardiomyopathic hearts, suggesting that depletion of energy reserve may contribute to the development of heart failure. PMID- 8732504 TI - Biochemical regulation of sarcoplasmic reticulum Cl- channel from human atrial myocytes: involvement of phospholamban. AB - Sarcoplasmic reticulum (SR) membrane vesicles derived from human atrium were characterized by specific ryanodine binding assay and fused into planar lipid bilayers. The tritiated form of the alkaloid bound to its receptor with a K(D) of 2.2 nM and a Bmax of 268 fmol/mg protein respectively. Special emphasis was placed on an anion-selective channel present in the SR membrane, which exhibited a mean conductance value of 67 pS when recorded in asymmetrical 50 mM trans/250 mM cis CsCl buffer system and a sensitivity to SITS (1 to 100 microM). Single and multiple channel activities displayed low voltage sensitivity and variability in its gating behavior which might result in spontaneous channel inactivation. However, the majority of the recordings (60%) resulted in a steady-state high open probability. The inactivated channel could be transiently reactivated with depolarizing voltage steps. This behavior is very similar, if not identical, to that observed for the SR Cl- channel in ventricular cells. The inactivation process is probably not directly related to a phosphorylation/dephosphorylation mechanism since PKA and PKG in presence of an adequate phosphorylation cocktail failed to reactivate the SR Cl- channel. In contrast, the use of a monoclonal anti-phospholamban antibody allowed the inhibition of the activity of the anionic channels. These results suggest that the regulation of the human atrial SR Cl- channel is dependent upon an interaction with phospholamban, which was clearly identified in our atrial preparations by Western blot analysis using monoclonal antibody. PMID- 8732505 TI - High affinity NAD(+)-dependent 11 beta-hydroxysteroid dehydrogenase in the human heart. AB - Receptor-ligand binding is an essential component of mineralocorticoid (MC) activity in target tissues. Detection of type 1 mineralocorticoid receptors (MR) in cardiac tissue is therefore suggestive that, like kidney, the heart is MC responsive. The presence of 11 beta-hydroxysteroid dehydrogenase (11 beta-HSD) within MC responsive tissue is essential to prevent saturation of MR by glucocorticoids. Using both high-performance liquid chromatography (HPLC) and thin layer chromatography (TLC), we have found that a high-affinity species of 11 beta-HSD predominates within human heart. Although two 11 beta-HSD isoforms were detected in human cardiac tissues, the activity of high-affinity (type 2) 11 beta HSD was found to be at least twice that of low affinity (type 1) 11 beta-HSD. Human cardiac type 2 11 beta-HSD possesses characteristics identical to the high affinity enzyme of distal renal tubules; 11 beta-dehydrogenation of corticosterone or cortisol to their 11-keto metabolites is NAD(+)-dependent and, with corticosterone as substrate, the enzyme has a nanomolar Km (15.1 nM as determined by Lineweaver-Burke analysis). Furthermore, its activity is unidirectional; corticosterone and cortisol are 11 beta-dehydrogenated to inactive 11-keto metabolites, whereas 11-oxoreductase activity (conversion of 11 dehydrocorticosterone and cortisone to corticosterone and cortisol, respectively) is absent. RT/PCR analysis, using primers complementary to the human renal type 2 11 beta-HSD sequence, demonstrated that the high-affinity species of 11 beta-HSD expressed in human heart is indeed the same enzyme as that produced in the kidney. These findings strongly suggest that, as is the case in the distal portion of the nephron, type 2 11 beta-HSD plays an important role in the human heart to promote glucocorticoid metabolism and to confer MC specificity upon MR. PMID- 8732506 TI - Cyclic AMP augments cytokine-stimulated nitric oxide synthesis in rat cardiac myocytes. AB - We investigated the effect of adenosine 3', 5'-cyclic monophosphate (cAMP) on inducible nitric oxide synthase (iNOS) expression in cultured neonatal rat cardiac myocytes. Incubation of cardiac myocytes for 24 h with interleukin-1 beta (IL-1 beta) caused a significant increase in the production of nitrite, a stable metabolite of nitric oxide. Dibutyl cAMP (db-cAMP) significantly augmented nitrite production by IL-1 beta-stimulated, but not by unstimulated cells, in a dose-dependent manner. db-cAMP also dose dependently increased nitrite production by tumor necrosis factor-alpha(TNF-alpha)-stimulated cells. Simultaneous incubation with NG-monomethyl-L-arginine completely inhibited the effect of db cAMP on nitrite production. cAMP-induced nitrite production by cytokine stimulated cells was accompanied by increased iNOS mRNA accumulation. The synergistic effect of cAMP on IL-1 beta-induced nitrite accumulation was mimicked by cAMP-generating agonists forskolin and isoproterenol. These results indicate that cAMP upregulates cytokine-induced iNOS expression in cardiac myocytes. PMID- 8732507 TI - Developmental differences and regional similarities in the responses of rat cardiac skinned muscles to acidosis, inorganic phosphate and caffeine. AB - The Ca2+ sensitivity of cardiac myofibrillar force production can be decreased by acidosis or inorganic phosphate (P(i)) and increased by caffeine. To investigate whether the source of tissue influences the potency of these agents, we compared the actions of acidosis (change of pH from 7.0 to 6.2), P(i) and caffeine (both 20 mM) on force production of skinned cardiac muscles from adult ventricle, adult atrium and neonate ventricle of the rat. Maximum Ca(2+)-activated force was reduced by all three interventions and the responses of the different muscle types to a given intervention were similar. Acidosis reduced myofibrillar Ca2+ sensitivity by 1.09 and 1.04 pCa units in adult ventricle and atrium, respectively, and P(i) reduced it by 0.19 and 0.22 pCa units. However, each effect was only one-third as great in the neonate ventricle, which showed falls of 0.33 pCa units for acidosis and 0.06 for P(i). In contrast, caffeine raised the Ca2+ sensitivity by the same amount (approximately 0.4 pCa units) in all three muscle types. The differential effect between adult and neonate seen with both acidosis and P(i) suggests some similarity in the mechanisms by which these factors decrease Ca2+ sensitivity. In contrast, the equal effects of caffeine on neonate and adult suggests that caffeine acts by a completely different mechanism. The lower pH- and P(i)-sensitivity of the neonatal ventricle can help to explain why neonatal and adult myocardium exhibit differential force responses to ischaemia (or hypoxia alone). PMID- 8732508 TI - Inhibition of L-type calcium current in rat ventricular cells by the tyrosine kinase inhibitor, genistein and its inactive analog, daidzein. AB - Effects of genistein, a specific inhibitor of tyrosine kinase, on the L-type Ca2+ channels were examined in freshly isolated young (days 10-18) rat ventricular cells using the whole-cell patch-clamp technique. Bath application of genistein decreased the L-type Ca2+ current [I(Ca(L))] in a concentration-dependent manner. The maximal inhibition of I(Ca(L)) was about 40% (attained at concentrations above 100 microM); the concentration for half-inhibition (IC50) was 11 microM. The effect of genistein (applied for about 5 min) was poorly reversible after washout for up to 5 min. The potential for half-inhibition (Vh) of the steady state inactivation curve was shifted in the negative direction by 7 mV (at 100 microM) by genistein, and the slope factor was also slightly changed; the activation curve was not affected. Daidzein, which is structurally related to genistein, but has little inhibitory effect on tyrosine kinase activity (of the EGF receptor), unexpectedly had almost the same inhibitory effect on I(Ca(L)). These observations suggest two possibilities for modulation of I(Ca(L)) in rat ventricular cells by genistein: (a) phosphorylation of the slow Ca2+ channels by tyrosine kinase; and (b) direct inhibition of the slow Ca2+ channels (i.e. independent of inhibition of tyrosine kinase activity). PMID- 8732509 TI - Sickle cell and thalassaemia: some priorities for nursing research. PMID- 8732510 TI - Pain management knowledge of hospital-based nurses in a rural Appalachian area. AB - The American Pain Society cites the failure of health care providers to routinely assess pain and pain relief as the most common reason for unrelieved pain in US hospitals. Lack of knowledge on the part of nurses about pain and its treatment is one of the major barriers to achieving comfort for those in pain. Thus, the purpose of this descriptive study was to determine the pain management knowledge of hospital-based nurses in a rural Appalachian area. The nonpurposive sample consisted of 123 registered nurses. The questionnaire consisted of three sections: demographic items and two instruments designed to measure knowledge of opioid classification and pain management. The overall pain management knowledge score was 67.4%. The findings indicate a knowledge deficit specifically related to the behavioural indicators of pain, classification of opioid analgesics, properties of opioid analgesics and adjuvant medications, and incidence of addiction. This sample of rural nurses demonstrates similar pain management knowledge compared to the reported scores of urban nurses. These results may serve as the impetus to develop strategies to decrease the barriers to effective pain management. Further education about pain management is warranted. PMID- 8732511 TI - Language and pain expression. AB - This paper arose during work for a BSc(Hons) dissertation, and considers the theoretical approach of Wittgenstein to pain analysis. This paper seeks to discuss one aspect of the research undertaken representing some of the findings which illustrate the association between pain and language. PMID- 8732512 TI - Stressors, coping mechanisms and quality of life among dialysis patients in Australia. AB - The aim of this study was to determine the significant stressors and coping methods which are related to quality of life in dialysis patients. A survey was conducted on patients in two dialysis centres in Sydney. The response rate was 58% (n = 64). The results revealed that limitation of physical activity was the most troublesome stressor followed by decrease in social life, uncertainty about the future, fatigue and muscle cramps. Although their ranking was not identical to that found in prior studies, these five stressors were considered to be compatible with the high stressors identified in previous studies. Problem solving methods were considered to be more effective than affective measures in dealing with stressors. Quality of life was perceived as below average in both haemodialysis and continuous ambulatory peritoneal dialysis (CAPD) patients. However, CAPD patients were experiencing a higher quality of life than haemodialysis patients. The length of time on the dialysis programme was not significantly related to coping behaviour. The findings of this study can further facilitate nurse practitioners in providing support, information, alternative solutions and in assisting patients to better utilize problem-solving methods to enhance their quality of life on dialysis. PMID- 8732513 TI - Helping the vulnerable or condoning control within the family: where is nursing? AB - The World Health Organization urges nurses to heed the health needs of the vulnerable groups 'to permit all citizens to lead socially and economically productive lives'. The characteristics of vulnerable adults and children are linked to relationship problems (Browne et al. 1988) within the family. Because of the diverse nature of families, it is not surprising that there is some ambiguity and complexity to the concept of vulnerability. This paper explores vulnerability within families through the explanations for domestic violence and child abuse; and concludes with the implications for nursing practice. PMID- 8732514 TI - Clinical guidelines: an introduction to their development and implementation. AB - There is a great deal of interest in the United Kingdom in clinical guidelines as a means of assisting practitioner and patient decision making about care options and in improving the quality of the care provided. Confusion remains, however, over what is meant by a clinical guideline and how it differs from and relates to protocols and standards. This paper was written under the auspices of the Royal College of Nursing Steering Group for the college's work on clinical guidelines, with the aim of clarifying some of the terminology used in the field and introducing ways in which clinical guidelines might be used by practitioners and patients to readers. At the moment just how effective the use of clinical guidelines can be on care is poorly established. What is known, however, is that crucial to their success are the strategies and methods used for their implementation. Such strategies and methods raise questions about how a sense of ownership can be engendered in those using the clinical guidelines and how they may be best operationalized. These questions are considered in this paper. PMID- 8732515 TI - Making sense of it: a neuro-interactional model of meaning emergence in critically ill ventilated patients. AB - Emphasis on meaning underpins a current thrust of knowledge development in nursing, especially in the client domain. Examination of meaning in the interactional context and through varying levels of consciousness has not been examined. Initially, an integrated model was developed deductively from philosophical, theoretical and research-oriented sources. This model was meant as a guide to begin examining how patients with varying levels of consciousness make sense of their intensive care unit experience. Over a 10-month period of fieldwork, this author observed patients twice daily through their intensive care unit stay to capture the nature and content of thinking processes. The resulting neuro-interactional model describes patients' thinking processes and scope of meaning as a function of levels of consciousness as well as factors which affect thinking and meaning. Theory, research and practice implications are presented. PMID- 8732516 TI - Non-necessary touch in the nursing care of elderly people. AB - This study describes the occurrence of non-necessary touching, defined as spontaneous and emotional physical contact between the nurse and patient, in the nursing care of elderly people in long-term care. The analysis was based on observation of nursing situations; any verbal communication occurring in connection with touching was also recorded. A total of 182 situations were studied in which 178 non-necessary touches by the nurse and four by the patient were recorded. Non-necessary touching was used in connection with a wide range of activities: making statements, making requests, waking up, encouraging, explaining, asking, comforting, teasing, telling off, hugging and thanking. PMID- 8732517 TI - Working with vulnerable families: a health visiting perspective. AB - The aim of this paper is to describe part of the findings of a research study which explored the health visitor's role in identifying and working with vulnerable families in relation to child protection. One objective of the study was to describe and analyse the work which health visitors undertake with vulnerable families aimed at preventing crisis, ill-health and child abuse. This paper outlines this aspect of the research and looks specifically at health visitors' perceptions of their work and interventions with vulnerable families and children. Health visitors working in an inner city area and a suburban area were sampled and pilot work was undertaken in a third suburban area. Stage one involved a postal survey of 102 health visitors, in which a response rate of 58 (57%) was obtained. Stage two involved 12 in-depth interviews with health visitors. One major finding of this study highlighted the fact that the health visitor's role when working with vulnerable families appears to be one of diversity and conflict. In the light of the current National Health Service reforms it seems of paramount importance that health visitors illustrate to others, both managers and other health/social service professionals alike, the important function of their work with these vulnerable groups. This is essential to ensure that health visitors' work with vulnerable families in relation to child protection is understood and valued and that the needs of vulnerable families continue to be identified. This paper contributes to the knowledge base of health visiting and raises some important issues for professional health visiting practice. PMID- 8732518 TI - Community psychiatric nurse education in the United Kingdom: 8 years of surveys and the issues raised. AB - Over the past 8 years annual surveys of the take-up and development of community psychiatric nursing courses within the United Kingdom have been conducted under the auspices of the Standing Advisory Group on Community Psychiatric Nurse Education (SAGCPNE). They reveal a clear link between changes in funding structure and numbers of community psychiatric nurses (CPNs) undertaking training, with recently implemented arrangements resulting in rises in student numbers. Rising numbers are not distributed evenly across course centres, with some courses showing more dramatic increases than others. As a consequence of Project 2000 and the UKCC PREP (Post-Registration Education and Practice policy), the surveys show that CPN courses have moved in many cases to diploma level and in some cases to degree level. A number of issues are raised by the changes charted by the surveys. Funding structures are set to change again within the next few years, and may have a negative impact upon CPN training. Market pressures have led to greater secrecy over curriculum content at a time when more openness could lead to shared improvements and greater uniformity of courses. In conjunction with these changes, government mental health policy has implications for CPN training. These issues are discussed and some modest recommendations made. PMID- 8732519 TI - The development of community psychiatric nursing: a professional project? AB - In this paper the author argues that the development of community psychiatric nursing can be understood as a professional project. However, the case of community psychiatric nursing cannot be adequately understood from mainstream literature on the subject of professionalization within the health care division of labour. The inadequacy of such literature is that it either concentrates upon the pre-eminent case of medicine or regards nursing's professionalization as being unitary. The author posits that the professional projects of general nursing and community psychiatric nursing have different and even opposing interests. To contextualize community psychiatric nurses' (CPNs') professional project the author examines: the history of mental nursing, from which community psychiatric nursing evolved; the development of community mental health care, which allowed for the inception of community psychiatric nursing; CPNs' relationships with both psychiatrists, whose patronage assisted CPNs' initial growth as para-professionals, and general practitioners, who later assisted a more functionally autonomous role for CPNs in primary care; and social policy towards community care that enhanced CPNs' growth. In conclusion the author argues that recent government-led changes in the management of community mental health care now impel CPNs to change the direction of their strategy for occupational survival and advancement. PMID- 8732520 TI - Primary health care team workshop: team members' perspectives. AB - This study explored members' perceptions of teamwork in two primary health care teams (PHCTs). It also examined the effect of a team-building intervention on members' perceptions centred around five topics: the PHCT, role perception, communication, leadership and conflict. The study used a qualitative approach with semistructured interviews before and after the intervention. It was found that members perceived each other's roles only in the light of their interactions with each other. Issues of hierarchy in leadership and interpersonal conflicts were raised. It is concluded that the team-building intervention had some positive effects on team members' perceptions and behaviour. However, further research is needed into management structures and conflict resolution in the PHCT. PMID- 8732521 TI - Promoting participation of informal caregivers in the hospital care of the elderly patient: informal caregivers' perceptions. AB - A study to identify informal caregivers' perceptions of factors promoting or inhibiting their participation in the care of elderly patients was conducted. An open-ended questionnaire was completed by 369 informal caregivers during 2-month periods in late 1991, 1992 and 1993. Several factors related to the patient, informal caregiver, nurse and environment were identified. The findings and implications for nursing are discussed. The data presented here are from a larger study which is reported in earlier issues of the Journal of Advanced Nursing. PMID- 8732522 TI - Kitwood's approach to dementia and dementia care: a critical but appreciative review. AB - This paper critically examines the recent work of Tom Kitwood and his colleagues at the Bradford Dementia Group. After initial details relating to Kitwood's early work and the development of the research group, the paper sets out Kitwood's ideas regarding the nature of dementia and dementia care. Four areas of contribution made by Kitwood are outlined. Problems relating to Kitwood's research methods are then put forward. In conclusion, it is suggested that while Kitwood's work will undoubtedly help enhance the care given to elderly demented people, more work needs to be done to ensure the validity of his methods. PMID- 8732523 TI - The biomedical construction of ageing: implications for nursing care of older people. AB - This paper reports on an existential phenomenological study carried out in a care of elderly people setting in a 1000-bed hospital in the United Kingdom. Fourteen participants were interviewed, each on several occasions. Two themes derived from these narratives are discussed, revealing negative experiences which are related to feelings of powerlessness. These two themes, routine geriatric style and segregation, are shown to arise from the history and culture of the wards and are shown to result in care deprivation and depersonalization. Patients' individual needs are ignored as they become the objects of inflexible routines within health care practice. In order to understand the situation, the history of care of older people and the biomedical construction of ageing are examined. It is concluded that what is needed is a wider social and political movement which opposes ageism and challenges ageist stereotypes. In addition, in health care there is a need for a review of the routine geriatric style of care and of segregation based on age and a social gerontology programme for nurse education. PMID- 8732524 TI - Caring, curing, coping: towards an integrated model. AB - There is an extensive literature discussing the "care:cure dilemma'. This usually puts forward the position that it is doctors who do the curing and nurses who do the caring. Patients are rarely included in the discussion. This paper considers some of this literature and examines whether this separation of functions is a valid one, the part of lay carers in the process, and the perspective of patients. It focuses particularly on cancer because this is an area where the issues seem particularly pertinent. The paper concludes by proposing an alternative and integrated conception which includes patients, nurses and doctors in a cognitive-phenomenological model of coping. It is suggested that this model moves forward from the previous limitations of the care:cure approach and enables more fruitful research and education of practitioners to be undertaken. PMID- 8732525 TI - Nurses' perceptions of irritable bowel syndrome (IBS) and sufferers of IBS. AB - The present study set out to discover nurses' attitudes towards, and beliefs about, irritable bowel syndrome (IBS) and IBS sufferers. A 54-item questionnaire was completed by 254 qualified nurses from 18 London hospitals. Nurses rated statements relating to IBS on a scale from disagree (1) to agree (5). Questionnaire items fell into four broad categories. The first related to the way nurses felt about the sufferers of IBS; the vast majority of nurses agreed with items stating that IBS patients are demanding, unable to cope with life, lazy and crave attention, and waste doctors' time. The second category related to nurses' beliefs regarding their knowledge of the syndrome; only half the sample believed that they would recognize the symptoms of IB and that they had a good understanding of the disorder. Ratings on items relating to the nurses' general knowledge of the symptoms showed that between a quarter and a half of the sample felt uncertain about the veracity of the statements. A fourth category was the nurses' beliefs regarding the attitudes held by health professionals, with half of the sample believing that doctors and health professionals generally had a poor understanding of IBS. Surprisingly, results showed that older, more experienced nurses, and those suffering from the syndrome were not more sympathetic and understanding than younger, non-sufferers. The present study shows that the majority of nurses hold negative attitudes towards IBS sufferers, which can only be detrimental to the treatment of those patients with IBS. PMID- 8732526 TI - The diagnosis of foetal distress: a challenge to midwives. AB - The diagnosis of foetal distress is complex: it is influenced by a number of factors which range from the physical examination of the mother and foetus to the psychological and social world in which the mother, the midwife and other health care professionals work. The consequences of an incorrect diagnosis of normality or foetal distress are potentially serious for both mother and baby. Recent changes in the practice and delivery of midwifery care have placed more responsibility on midwives. It is therefore imperative that midwives become more aware of the complexity of the diagnosis as well as of some of the factors which can improve the quality of their diagnosis. This paper reviews the literature associated with the diagnosis of foetal distress, physiological and pathological changes during pregnancy and labour, and the usefulness and limitation of the tools readily available to midwives for the diagnosis. It also examines the limits of the relationship that exists between intra-uterine and neonatal markers. The purpose of this paper is to provide background information that would support midwives in the diagnosis of foetal distress in the hope that the false positive diagnoses can be reduced for the benefit of mothers and babies. Ultimately an improvement in outcome should be reflected in the more efficient and effective use of resources available to maternity services. PMID- 8732527 TI - The women's health agenda: evolution of hormone replacement therapy as treatment and prophylaxis for coronary artery disease. AB - The purpose of this paper is to develop an understanding of the research literature on the connection between hormone replacement therapy and development of coronary artery disease as it pertains to women. It is clear that research in this century regarding heart disease in general has been gender-biased or inappropriately gender-neutral. Research initially focused on how the developing knowledge of the protective effect of female hormones could be used to benefit men. It has only been in the last decade or so that gender-based research has been appreciated, acknowledged, facilitated, and finally accepted for its contributions to advancing knowledge and for benefiting women with coronary artery disease. PMID- 8732528 TI - Nursing students' experiences caring for cognitively impaired elderly people. AB - Over the next half century the population of moderately to severely demented individuals is projected to nearly triple. Because high complex technology is usually not required in the care of cognitively impaired elderly people, these individuals may be the first patients nursing students are assigned to care for. Despite the fact that these elderly people do not require "high tech' care, their nursing care can be extremely challenging for nursing students. In order to help prepare nursing students to meet this challenge, a phenomenological study was conducted to describe the meaning of nursing students' experiences caring for cognitively impaired elderly people. Thirty-seven undergraduate nursing students participated in the study. Each student described in writing an experience he/she had had while caring for a cognitively impaired elderly person. Using Colaizzi's phenomenological method to analyse these descriptions, five themes emerged. Nursing students experienced a myriad of emotions such as frustration, sadness, fear and empathy. Caring for cognitively impaired elderly people presented difficult challenges for nursing students to overcome. Because of the difficulties encountered in caring for confused elderly people, nursing students utilized multiple nursing care approaches. As a result of giving care to elderly people suffering from dementia, negative consequences ensued for some nursing students. The positive outweighed the negative consequences in the phenomenological study. Implications for nurse educators are addressed. PMID- 8732529 TI - Participation in continuing professional education in nursing: findings of an interview study. AB - The evidence suggests that enrolled nurses and those working part-time and on night duty consistently attend less continuing professional education than their more senior, full-time and day duty colleagues. This was substantiated in the findings of this study which also highlighted the paucity of opportunities for continuing education among unqualified practitioners. In the light of recent developments within the health services, this inequitable provision of continuing professional education to all nursing practitioners will need to be addressed and resolved. Continuing professional education provided on an arbitrary and random basis will not afford maximum positive outcomes for clients, the service and practitioners and has the potential to increase the levels of frustration among disadvantaged groups of nursing staff. PMID- 8732530 TI - Quantitative methods in nursing research. AB - This paper considers some of the general principles that are common to much of quantitative research in nursing. In particular, the role of hypothesis testing is considered, and the use of estimation is emphasized. Confidence intervals are advocated as a means of assessing both statistical and clinical significance. These principles and methods are illustrated using examples taken from the nursing literature. Finally, more complicated situations and the role of design are discussed. PMID- 8732531 TI - A quasi-experimental research to investigate the retention of basic cardiopulmonary resuscitation skills and knowledge by qualified nurses following a course in professional development. AB - The research was undertaken with the intention of testing six null hypotheses regarding the retention of basic cardiopulmonary resuscitation (CPR) skills and knowledge of registered nurses. The hypotheses were formulated from the broad aims of the research, which were to investigate conclusions reached by other researchers highlighting the speed with which retention of CPR skills and knowledge deteriorates, and to investigate the need for regular updating in CPR. The research was quasi-experimental in nature. The 19 nurses participating in the research were qualified staff undertaking the English National Board (ENB) 923 course in Professional Development, which included a refresher on basic CPR skills and included some discussion regarding advanced techniques. The latest guidelines issued by the Resuscitation Council (1993) were used, which also aided in the design and use of the two research tools, namely an eight-point skills testing observation tool and a 26-point knowledge-testing questionnaire. While a 3-hour update in CPR skills revealed an initial improvement, the decrease in retention of skills 10 weeks later was significant (P = 0.0000). The update in CPR knowledge also revealed an initial improvement but the decrease in retention of knowledge 10 weeks later was significant (P = 0.0000). The findings of the research reflect similar results to previous research undertaken and discussed in the literature review, suggesting that retention of skills and knowledge quickly deteriorates if not used or updated regularly. Therefore this research supports the importance of CPR refresher courses on a regular basis. PMID- 8732532 TI - The use of the Nominal Group Technique in research with community nurses. AB - The use of group-based techniques to generate ideas and offer solutions to problems is commonplace in many disciplines and for many purposes. What were originally useful adjuncts to market research have become valuable and illuminating tools in education, policy and research. This paper will describe the use of one such technique, the Nominal Group Technique, and its practical application in bridging the gap between researchers and practitioners in one particular study of community nursing. PMID- 8732533 TI - Nursing theory: everything the artist spits is art? AB - This paper explores the concept and utility of nursing theory in and for the practice of nursing. Working from the premise that many nurse practitioners appear uncertain as to the value of theory in relation to their everyday working experience, the paper investigates the contribution nursing theory makes in terms of sustaining and developing nursing as a practice discipline. The fact that nursing theory remains at once poorly evaluated, articulated or understood appears to be compounded by a general perception of nurse theorists as being removed from the realities of the practice setting and by the confusion precipitated, not least, by the semantic ambiguity engendered by their writings. The paper reviews the complex relationships extant between the development of nursing theory in regard to its utility for nursing practice, and concludes by suggesting a practice-led perspective by which nursing theory may be better articulated and assimilated within the discipline. In order to facilitate the study, it has been necessary to investigate the historical, theoretical and philosophical imperatives pertaining not only to the development of nursing theory but to nursing research and nursing practice per se. PMID- 8732534 TI - The practitioner-teacher: a study in the introduction of mentors in the preregistration nurse education programme in Wales: Part 1. AB - This study focused on the introduction of mentors in the Common Foundation Programme of Project 2000 schemes of preregistration nurse education. The research, which was commissioned by the Department of Health Research and Development Division on behalf of the Welsh Office Nursing Division, began in February 1992 and was undertaken on an all-Wales basis. The completed report aims to provide policy makers with information relating to important issues which are central to the teaching and learning of nursing in clinical locations. This first paper discusses some background issues and gives a brief conceptual framework for considering policy reform, summarizes the research questions which emerged and describes the methods used to address them. The second paper (to appear in the next issue of the journal) will describe the key findings from the study and discuss some ensuing potential implications and considerations for all those involved in the preparation of future practitioners of nursing. PMID- 8732535 TI - Health care and nursing in Romania. AB - Health care and the nursing profession have been affected dramatically by political and socio-economic conditions in Romania. The Romanian Ministry of Health has recognized the health problems and barriers to care delivery which exist in the country, with several goals identified to improve the status of Romanian health care. This paper explores the critical issues which have impacted on health care and nursing in Romania. PMID- 8732536 TI - Concept of grief. PMID- 8732537 TI - Ionotropic glutamate receptors. Their possible role in the expression of hippocampal synaptic plasticity. AB - In the brain, most fast excitatory synaptic transmission is mediated through L glutamate acting on postsynaptic ionotropic glutamate receptors. These receptors are of two kinds--the alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA)/kainate (non-NMDA) and the N-methyl-D-aspartate (NMDA) receptors, which are thought to be colocalized onto the same postsynaptic elements. This excitatory transmission can be modulated both upward and downward, long-term potentiation (LTP) and long-term depression (LTD), respectively. Whether the expression of LTP/LTD is pre-or postsynaptically located (or both) remains an enigma. This article will focus on what postsynaptic modifications of the ionotropic glutamate receptors may possibly underly long-term potentiation/depression. It will discuss the character of LTP/ LTD with respect to the temporal characteristics and to the type of changes that appears in the non-NMDA and NMDA receptor-mediated synaptic currents, and what constraints these findings put on the possible expression mechanism(s) for LTP/LTD. It will be submitted that if a modification of the glutamate receptors does underly LTP/LTD, an increase/ decrease in the number of functional receptors is the most plausible alternative. This change in receptor number will have to include a coordinated change of both the non-NMDA and the NMDA receptors. PMID- 8732538 TI - Physiological relevance and functional potential of central nervous system derived cell lines. AB - Central nervous system (CNS)-derived neural cell lines have proven to be extremely useful for delineating mechanisms controlling such diverse phenomena as cell lineage choice and differentiation, synaptic maturation, neurotransmitter synthesis and release, and growth factor signalling. In addition, there has been hope that such lines might play pivotal roles in CNS gene therapy and repair. The ability of some neural cell lines to integrate normally into the CNS following transplantation and to express foreign, often corrective gene products in situ might offer potential therapeutic approaches to certain neurodegenerative diseases. Five general strategies have evolved to develop neural cell lines: isolation and cloning of spontaneous or mutagenically induced malignancies, targeted oncogenesis in transgenic mice, somatic cell fusion, growth factor mediated expansion of CNS progenitor or stem cells, and retroviral transduction of neuroepithelial precursors. in this article, we detail recent progress in these areas, focusing on those cell lines that have enabled novel insight into the mechanisms controlling neuronal cell lineage choice and differentiation, both in vitro and in vivo. PMID- 8732539 TI - Neurotrophin-3 as an essential signal for the developing nervous system. AB - Rapid advances in characterization of the biological actions mediated by the third member of the neurotrophin family, neurotrophin-3 (NT-3), have been made recently in vitro as well as in situ. These have been made possible by the cloning of the genes for NT-3 and for its transducing receptor tyrosine kinase TrkC. This article will focus on the roles of NT-3 in the nervous system. In situ localization of NT-3 consistent with that of its receptor is manifested at all developmental stages studied and into adulthood. Through TrkC, NT-3 signals a number of trophic effects, ranging from mitogenesis, promotion of survival, or differentiation, depending on the developmental stage of the target cells. The sites of action of NT-3 reside primarily in the peripheral nervous system (PNS), various areas of the central nervous system (CNS), and in the enteric system (ENS). Analyses of the phenotypes of transgenic mice lacking NT-3 or injection of embryos with a blocking antibody have so far revealed the essential role of NT-3 in development of specific populations of the PNS, and in particular of proprioceptive, nodose, and auditory sensory neurons and of sympathetic neurons. The actions of NT-3 also extend to modulation of transmitter release at several types of synapses in the periphery as well as in the adult CNS. In addition, NT-3 may play a role in the development of tissues other than the nervous system, such as the cardiovascular system. Future investigations will widen the understanding of the many roles of NT-3 on both neuronal and nonneuronal cells. PMID- 8732542 TI - Unique growth properties of neonatal pulmonary vascular cells: importance of time and site-specific responses, cell-cell interaction, and synergy. PMID- 8732540 TI - Brain GABAA receptors studied with subunit-specific antibodies. AB - Brain GABAA/benzodiazepine receptors are highly heterogeneous. This heterogeneity is largely derived from the existence of many pentameric combinations of at least 16 different subunits that are differentially expressed in various brain regions and cell types. This molecular heterogeneity leads to binding differences for various ligands, such as GABA agonists and antagonists, benzodiazepine agonists, antagonists, and inverse agonists, steroids, barbiturates, ethanol, and Cl- channel blockers. Different subunit composition also leads to heterogeneity in the properties of the Cl- channel (such as conductance and open time); the allosteric interactions among subunits; and signal transduction efficacy between ligand binding and Cl- channel opening. The study of recombinant receptors expressed in heterologous systems has been very useful for understanding the functional roles of the different GABAA receptor subunits and the relationships between subunit composition, ligand binding, and Cl- channel properties. Nevertheless, little is known about the complete subunit composition of the native GABAA receptors expressed in various brain regions and cell types. Several laboratories, including ours, are using subunit-specific antibodies for dissecting the heterogeneity and subunit composition of native (no reconstituted) brain GABAA receptors and for revealing the cellular and subcellular distribution of these subunits in the nervous system. These studies are also aimed at understanding the ligand-binding, transduction mechanisms, and channel properties of the various brain GABAA receptors in relation to synaptic mechanisms and brain function. These studies could be relevant for the discovery and design of new drugs that are selective for some GABAA receptors and that have fewer side effects. PMID- 8732543 TI - Alterations in extracellular matrix protein expression in neonatal pulmonary hypertension: implications for vascular function. PMID- 8732544 TI - Mechanisms of abnormal vasoreactivity in persistent pulmonary hypertension of the newborn infant. PMID- 8732541 TI - Glutamate and Parkinson's disease. AB - Altered glutamatergic neurotransmission and neuronal metabolic dysfunction appear to be central to the pathophysiology of Parkinson's disease (PD). The substantia nigra pars compacta--the area where the primary pathological lesion is located- is particularly exposed to oxidative stress and toxic and metabolic insults. A reduced capacity to cope with metabolic demands, possibly related to impaired mitochondrial function, may render nigral highly vulnerable to the effects of glutamate, which acts as a neurotoxin in the presence of impaired cellular energy metabolism. In this way, glutamate may participate in the pathogenesis of PD. Degeneration of dopamine nigral neurons is followed by striatal dopaminergic denervation, which causes a cascade of functional modifications in the activity of basal ganglia nuclei. As an excitatory neurotransmitter, glutamate plays a pivotal role in normal basal ganglia circuitry. With nigrostriatal dopaminergic depletion, the glutamatergic projections from subthalamic nucleus to the basal ganglia output nuclei become overactive and there are regulatory changes in glutamate receptors in these regions. There is also evidence of increased glutamatergic activity in the striatum. In animal models, blockade of glutamate receptors ameliorates the motor manifestations of PD. Therefore, it appears that abnormal patterns of glutamatergic neurotransmission are important in the symptoms of PD. The involvement of the glutamatergic system in the pathogenesis and symptomatology of PD provides potential new targets for therapeutic intervention in this neurodegenerative disorder. PMID- 8732545 TI - Clinical pathophysiology of persistent pulmonary hypertension of the newborn and the role of inhaled nitric oxide therapy. AB - Inhaled NO can cause marked improvement in oxygenation when optimal lung inflation is achieved and systemic blood volume and vascular resistance are adequate. However, inhaled NO alone is unlikely to cause sustained improvements in oxygenation in neonatal hypoxemic respiratory failure associated with severe parenchymal lung disease without extrapulmonary shunting. Inhaled NO may be an important tool in the management of severe PPHN when its application is limited to patients with severe extrapulmonary shunting. PMID- 8732546 TI - Neonatal respiratory disease due to surfactant protein B deficiency. PMID- 8732547 TI - Cardiopulmonary consequences of congenital diaphragmatic hernia. AB - The cardiopulmonary consequences of CDH can be devastating, and despite intensive research the pathophysiologic makeup of this condition is not fully understood. It seems likely that its overall prognosis is related to the size of the pulmonary vascular bed and its capacity to accept the cardiac output at birth. The ability of the lungs to perform this task is impaired by a surfactant deficiency and by pulmonary vessels that respond excessively to normal stimuli of vasoconstriction. Understanding the mechanisms through which these pathophysiological abnormalities are occurring should provide us with therapeutic insights into how we may recruit all the available alveoli in these lungs, prevent ventilation-induced lung injury, optimize pulmonary blood flow, and maximize cardiac output without causing right or left heart compromise. Failure to achieve all or any of these goals can feed back and exacerbate the underlying pathophysiological condition. PMID- 8732549 TI - Physiology and clinical role of liquid ventilation therapy. PMID- 8732548 TI - "State-of-the-art" management strategies for the fetus and neonate with congenital diaphragmatic hernia. PMID- 8732550 TI - Clinical approaches to the use of high-frequency oscillatory ventilation in neonatal respiratory failure. AB - Hypoxemic respiratory failure in the newborn infant may be caused by underinflation and intrapulmonary shunting often associated with pulmonary hypertension and concomitant extrapulmonary shunting. HFOV can be an effective tool to recruit and sustain lung volume in this setting. PMID- 8732551 TI - Neonatal ventilator management and the role for pulmonary function tests. PMID- 8732552 TI - Managed care teamwork: perinatal bundling. PMID- 8732553 TI - Comparison of upper arm and calf oscillometric blood pressure measurement in preterm infants. AB - Although the practice of using calf blood pressure (BP) measurement is widespread, virtually no information is available about calf site BP measurements in preterm infants. This research compared oscillometric upper arm BP measurements with calf BP measurements in 65 preterm infants with a mean weight of 1629 +/- 437 gm. Differences between arm and calf BP measurements were analyzed with repeated-measures multivariate analysis of variance of systolic, diastolic, and mean arterial pressures. There were no significant differences between arm and calf systolic BP on days 1, 3, and 5 of life. With post hoc testing there was a significant difference on day 7 between systolic arm BP (60.4 +/- 8.4 mm Hg) and calf BP (57.7 +/- 7.1 mm Hg) (t = 3.68, p < 0.01). There were no significant differences between arm and calf measurements for diastolic and mean arterial pressures and no main effect from infant weight on differences between arm and calf BP. It may be reasonable to use either arm or calf BP during the first 5 days of life. PMID- 8732554 TI - Adverse perinatal outcome in the older primipara. AB - Delayed childbearing has become common and has raised the awareness of the possible risks for the mother and the newborn infant. The increased maternal and neonatal risks have been attributed largely to the lack of proper prenatal care. The aim of this study was to assess whether advanced maternal age is a significant risk factor in mothers who receive good prenatal care. We matched 161 cases 1:1 according to the following criteria: maternal and paternal ethnic origin, chronic diseases, marital status, and smoking during pregnancy. Our results show that the older women had babies with a significantly higher incidence of low birth weight (< 2500 gm, p = 0.001), prematurity (< 37 weeks, p = 0.02), intrauterine growth retardation (p = 0.001), abruptio placentae (p = 0.002), and cesarean section (p < 0.001). The average hospital stay for the babies of the older mothers was longer than that for babies of the younger mothers (8.4 vs 6.1 days, p = 0.003), and the incidence of hospitalization for more than 3 days in the neonatal intensive care unit was increased (10.3% vs 2.2%). Logistic regression did not support maternal age of 35 years and older as being the single significant risk factor for adverse neonatal and maternal outcome. We conclude that maternal age older than 35 years entails a higher risk for the mother and her newborn infant, even when good prenatal care is taken. PMID- 8732555 TI - Tolerance of initial diphtheria-tetanus-pertussis immunization in preterm infants. AB - With the use of neurologic examinations, cranial sonograms, electroencephalograms, an cry analyses, we assessed neurologic function before and after an initial diphtheria-tetanus-pertussis immunization in 22 very low birth weight infants. Mean birth weight was 1036 +/- 137 gm; mean gestational age was 28.0 +/- 1.3 weeks. All 22 infants had recovered from respiratory distress syndrome, 10 infants had been treated for invasive bacterial or fungal infection, and 9 infants had had previous intraventricular hemorrhages. Initial immunization was administered at a mean age of 71 +/- 15 days (range 57 to 120 days) and a mean weight of 1895 +/- 245 gm (1370 to 2280 gm). Clinical reactions were mild and transient. No postimmunization changes in neurologic examinations or objective studies were noted compared with results of preimmunization studies. These findings support the safety of administering an initial diphtheria-tetanus pertussis immunization to very low birth weight infants at the recommended age of 8 weeks. PMID- 8732556 TI - Cervical assessment: visual or digital? AB - Our hypothesis for this study was that visual cervical assessment is not equivalent to digital assessment in patients with uterine contractions. We compared visual with digital cervical examinations done on 133 gravid women evaluated in the labor and delivery department because of uterine contractions. Patients at 27 weeks' gestation or more, subjectively in early labor, were prospectively entered into the study. Cervical dilatation and effacement were determined by two separate examiners blinded to each other's assessment, one by digital examination and the other by visual examination, in random order no more than 5 minutes apart. Eight of 133 patients were excluded from data analysis because of inability to visualize the cervix during speculum examination. Data analysis was done with correlation coefficient and the Bland and Altman test for agreement of two clinical measurements. Visual examination only underestimated actual cervical dilation by 0.60 cm (95% confidence interval [CI] 0.58 to 0.62 cm) and the limits of agreement were -2.06 cm (95% CI -2.02 to -2.09 cm) to 3.25 cm (95% CI 3.21 to 3.29 cm). Similarly, visual assessment underestimated effacement by 14.2% (95% CI 13.7% to 14.6%) and the limits of agreement were 41.3% (95% CI -40.5% to 42.0%) to 69.9% (95% CI 68.8% to 70.4%). In conclusion, cervical assessment in patients with uterine contractions who are thought to be in labor by visual speculum examination is not equivalent to that by digital examination. This difference seems to only be clinically significant if the cervix is more than 3 cm dilated. PMID- 8732557 TI - Infant care knowledge of primiparous urban mothers. AB - Because hospitals are discharging postpartum women within 48 hours of delivery, little time is available for maternal baby care instruction. Therefore it is important that any teaching, whether in the hospital or in an outpatient setting, concentrate on topics that provide the greatest benefit to the mother and child. This study provides an initial view of maternal knowledge of well baby care in an attempt to discern topics of common misinformation and misunderstanding. Fifty primiparous, English-speaking mothers were interviewed. The interview consisted of questions dealing with social support for the mother; employment or schooling; source of prior child care knowledge; and knowledge of feeding, diapering, sleeping, calming, bathing, and sickness of newborn infants. The data gathered indicate that although most women report one or more sources of knowledge, several gaps exist in their preparedness. The traditional information presented may overemphasize known areas at the expense of some that have significant consequences for health and safety. Many hospitals offer postpartum classes in bathing. These data indicate that if these programs could be expanded to include more practical information, they would provide a beneficial learning experience at a critical time in a new mother's life. PMID- 8732558 TI - Feeding of premature infant formula after hospital discharge of infants weighing less than 1800 grams at birth. AB - A randomized, double-blind study was conducted to determine whether continued feeding of premature infant formula after hospital discharge improve biochemical measures of bone mineral or protein status and anthropometrics during the first 8 and 12 weeks, respectively, after initial hospital discharge. Forty-three subjects were randomized to receive either a 20 kcal/ounce standard cow's milk based formula with iron or a 20 kcal/ounce premature infant formula with iron for 8 weeks after hospital discharge. Sixteen exclusively breast-fed infants (mother's own milk) who received a multivitamin supplement with iron were compared with infants in both formula groups. There were no differences among the three groups in gender, birth weight, gestational age, or weight and age at the time of study entry. Alkaline phosphatase values were lower in infants receiving premature infant formula than in those receiving standard formula 8 weeks after discharge. Phosphorus values were lower and alkaline phosphatase values higher in the human milk-fed group than in both formula groups 8 weeks after discharge despite supplementation with calcium, phosphorus, and vitamin D before and during the study. At 8 weeks after discharge, human milk-fed infants also had lower transferrin levels than infants fed formulas. Infants in both formula groups grew similarly in weight, whereas the infants fed human milk weighed less throughout the study. The group fed premature infant formula had greater mean length and head circumference than the standard formula or human milk-fed groups. These data indicate that premature infants weighing < 1800 gm at birth may benefit from the continuation of premature infant formula during the first 8 weeks after initial hospital discharge. PMID- 8732559 TI - A normative study of obstetric complications associated with fetal trisomy 18. AB - Our objective was to determine whether the cesarean section rate and amniotic fluid lecithin-sphingomyelin ratio (L/S ratio) associated with fetal trisomy 18 are different from those associated with euploid pregnancies. Forty-nine trisomy 18 pregnancies were identified and their records were reviewed. Of the 22 live born infants, 15 (68%) were delivered by cesarean section. The overall cesarean section rate for the institution ranged from 19.1% to 22.6%. In five patients with gestational ages between 35 and 38 weeks who had amniocentesis as part of the evaluation of intrauterine growth retardation, the L/S ratios were < or = 1.8. These data confirm that undiagnosed trisomy 18 pregnancies are associated with an increased cesarean section rate. Further, trisomy 18 fetuses appear to have delayed maturation of the L/S ratio. These results reinforce the importance of a karyotypic evaluation of selected pregnancies complicated by intrauterine growth retardation, even in the third trimester, and suggest that the prenatal diagnosis of trisomy 18 is of obstetric importance. PMID- 8732560 TI - Cardiac arrest in an extremely low birth weight infant: complication of percutaneous central venous catheter hyperalimentation. AB - We report a case of unexpected sudden cardiac arrest in an extremely low birth weight infant on the eighth day after a percutaneous silicone rubber central venous catheter was inserted for parenteral nutrition. The unique pathologic condition of infiltration of the myocardium with probably intralipid and degeneration of myocardial fibers over the right atrium and part of the right ventricle has not been reported in the past. There was no pericardial effusion. A brief review of the complications of central venous catheters is given. PMID- 8732561 TI - Nitroglycerin facilitates therapeutic cerclage placement. AB - Therapeutic cerclage placement may be complicated by prolapsing of the fetal membranes. A gravida presented with incompetent cervix and prolapse of fetal membranes. The membranes were not reduced by Trendelenburg position, decompressive amniocentesis, and spinal anesthesia. Intravenous nitroglycerin promptly reduced the prolapse and allowed cerclage placement. This is the first report of intravenous nitroglycerin tocolysis used to facilitate cerclage placement. PMID- 8732562 TI - Inferior vena cava catheterization in the neonate by the percutaneous femoral vein method. AB - Placement of percutaneous femoral venous catheters in neonates is rarely performed for long-term central venous access. We studied 44 neonates (gestational age, 32.4 +/- 48 weeks, postnatal age, 18 +/- 32 days) with lack of other venous access sites. The femoral vein was entered with a 19-gauge introducer needle with placement of a 2F silastic catheter localized at the inferior vena cava-right atrial junction. Catheter-related sepsis, bacteremia, or thrombophlebitis did not occur in any patients. Our findings suggest that percutaneous femoral-vena cava catheters can be safely used when other nonfemoral sites are unavailable. PMID- 8732563 TI - Decision making for the critically ill neonate near the end of life. AB - Our purpose was to review the medical records of 122 infants who died in our neonatal intensive care unit (NICU) to characterize decision making for the critically ill neonate near the end of life. The majority of deaths (72%) were related to some complication of prematurity. Families participated in the decision-making process in 75% of the instances. A decision was made to limit, withdraw, or withhold life-sustaining treatment for 82% of the infants. At the time of decision making, prognosis was judged to be poor or hopeless and the burdens of treatment unacceptable for 90% of the infants. This study confirms that health care providers and families together can confront the ethical decision of whether to continue or forego life-sustaining treatment for the critically ill neonate near the end of life and, in the majority of cases, will choose to limit, withdraw, or withhold such support. PMID- 8732564 TI - Antenatal-onset infantile cortical hyperostosis and nonimmune hydrops. AB - Infantile cortical hyperostosis antenatal onset is an uncommon disease characterized by polyhydramnios, anasarca or hydrops, pulmonary hypoplasia, hepatomegaly, bowed hyperostotic long bones, and a poor prognosis. Sonographically the intrauterine manifestations may be similar to those of osteogenesis imperfecta, type II. PMID- 8732565 TI - Subcutaneous fat necrosis of the newborn: a case presentation. AB - Subcutaneous fat necrosis is normally a self-limiting condition. A complication of subcutaneous fat necrosis is hypercalcemia. If the infant is discharged from the hospital before the lesions are healed, the family should be instructed to monitor the infant for the signs and symptoms of hypercalcemia. The parents need to understand the importance of keeping follow-up appointments to monitor the calcium levels. It is important to reassure the family that the lesions will disappear with time. They need to be instructed to leave the lesions alone. The lesions should not be punctured, nor should the family apply hot or cold compresses to the sites. The family should be cautioned against exposing the baby to sunlight to lower endogenous vitamin D production. PMID- 8732566 TI - Neonatal radiology casebook. Massive systemic air embolism in a low birth weight infant. PMID- 8732567 TI - Special imaging casebook. Meckel-Gruber syndrome. PMID- 8732568 TI - Pleural effusion caused by intrathoracic central venous hyperalimentation. PMID- 8732569 TI - Adequacy of antenatal phenobarbital dosing for neonatal intraventricular hemorrhage prophylaxis. PMID- 8732570 TI - Managed care and neonatology. PMID- 8732571 TI - Controlled temperature tissue fusion: argon laser welding of canine intestine in vitro. AB - BACKGROUND AND OBJECTIVE: Thermal denaturation of proteins is recognized as a rate process governed by the local temperature-time response and is believed to be the principal mechanism for photothermal tissue welding. Since rate processes are exponential with temperature, feedback control of tissue surface temperature is hypothesized to create a quasi-constant rate of denaturation that will enhance the tissue welding process. STUDY DESIGN, MATERIALS AND METHODS: Controlled temperature tissue welding of severed edges of fresh canine jejunum was performed in vitro by remote sensing of tissue surface temperature with an infrared sensor. A hardware controlled temperature feedback system opened and closed a shutter located in the beam path of an argon ion laser to provide constant temperature welding. RESULTS: Strong tissue fusion was not possible at or below a surface temperature of 70 degrees C, but was accomplished at 80 degrees, 90 degrees, and 95 degrees, and 100 degrees C. Fusion was achieved with thermal coagulation of the collagenous submucosa and mucosal tissues. The bursting strength of welds created at 90 degrees C and 95 degrees C were significantly stronger than those performed at 80 degrees C. CONCLUSION: Laser-assisted intestinal anastomoses created in vitro are optimally strong at 90-95 degrees C feedback control temperatures. PMID- 8732572 TI - Temperature-controlled laser photocoagulation of soft tissue: in vivo evaluation using a tissue welding model. AB - BACKGROUND AND OBJECTIVES: Laser surgical procedures involving photocoagulation of soft tissue have relied on subjective visual endpoints. The thermal damage to the denatured tissue in these procedures is highly dependent on the tissue temperatures achieved during laser irradiation. Therefore, a system capable of real time temperature monitoring and closed loop feedback was used to provide temperature controlled photocoagulation (TCPC). STUDY DESIGN/MATERIALS AND METHODS: The TCPC system consisted of a 1.32 microns Nd:YAG laser, an infrared thermometer, and a microprocessor for data acquisition and feedback control. A porcine skin model was used. Tissue welds were completed to evaluate the photocoagulation effects at different predetermined temperatures. A quantitative measurement of tissue photocoagulation was obtained by tensile strength measurements of the laser repairs. Histology of the irradiated tissue was used to determine the extent of thermal injury associated with different photocoagulation temperatures. RESULTS: The TCPC system was capable of maintaining a relatively constant temperatures (+/- 4 degrees C) during laser irradiation. The tensile strengths of acute repairs increased with temperature over the range studied (65 95 degrees C). Tensile measurements made after several days of healing showed that higher temperature (95 degrees C) welds had lower strengths than repairs completed at lower (65 degrees C or 75 degrees C) temperatures and were significantly lower at 3 days. Acute histology showed that the amount thermal damage was strongly dependent on the tissue temperature and increased both in tissue depth and lateral to the repair with temperature. The histologic results suggest that the increase in the acute repair tensile strength as the weld temperature increased was due to an increase in the depth of tissue photocoagulation. The increase in the lateral tissue injury measured histologically for higher temperature welds likely resulted in the decreased chronic tensile strengths, as a healing response to excessive thermal damage. CONCLUSION: Tissue temperatures can be controlled during laser photocoagulation of skin. The degree of acute and chronic tissue damage is highly dependent on the temperature during welding. By controlling the tissue temperature during laser procedures, the surgical outcome can be more reliably predicted and reproduced, as compared to the conventional open loop methods. In addition, the use of a TCPC system should significantly reduce the learning curve for photothermal surgical procedures. PMID- 8732573 TI - Light distributions in a port wine stain model containing multiple cylindrical and curved blood vessels. AB - BACKGROUND AND OBJECTIVES: Knowledge of the light distribution in skin tissue is important for the understanding, prediction, and improvement of the clinical results in laser treatment of port wine stains (PWS). The objective of this study is to improve modelling of PWS treated by laser using an improved and more realistic PWS model. STUDY DESIGN/MATERIALS AND METHODS: Light distributions are calculated by the Monte Carlo method for various PWS blood vessel configurations, such as single and multiple vessels oriented horizontally, curved vessels, and "vertically" oriented vessels. Various vessel sizes and wavelengths are used. RESULTS: Our modelling confirms the concept of selective photothermolysis; 577nm laser light gives maximal deposited energy at the top side of the blood vessels closest to the skin surface and 585nm gives a more uniform energy distribution in the vessel. In the distribution of deposited energy multiple vessels mutually influence each other, because of "shadowing" of diffuse light. CONCLUSION: Modelling PWS laser treatments with multiple vessels confirms the need for successive treatments of vessels layer by layer. The use of different wavelengths affects the local deposited energy profiles in the blood vessels. It predicts that the significance of 585nm laser light lies in the uniform energy distribution in the vessels rather than in gain in energy deposition with depth. The calculated light distributions provide a more realistic input for modelling thermal damage effects in PWS laser treatment and modelling of the epidermal response in thermal imaging of the PWS blood vessel structure. PMID- 8732574 TI - Histological responses of port wine stains in brown skin after 578 nm copper vapor laser treatment. AB - BACKGROUND AND OBJECTIVES: The object of this study is to characterize the effects of epidermal melanin in brown skin on selective vessel damage by copper vapor laser radiation in port wine stain (PWS). STUDY DESIGN/MATERIALS, AND METHODS: We observed the histological changes of PWS in Korean patients who received copper vapor laser (578 nm) treatment over a range of energy densities (6-14 J/cm2) and exposure durations (30-200 ms). The nitroblue tetrazolium chloride (NBTC) staining method was used to differentiate between the blue stained viable cells and the unstained thermally damaged cells. RESULTS: With Fontana-Masson stain, we found that Korean skin has more epidermal melanin than Caucasian skin. For energy densities greater than 6 J/cm2, epidermal damage was observed. At 6 and 8 J/cm2, the damage to the dermis was localized to the blood vessels and the perivascular tissue. The connective tissue between damaged vessels and epidermis was still viable. Energy densities above 10 J/cm2 produced a diffuse thermal necrosis. We conclude that vascular selectivity without epidermal damages cannot be achieved with a 50 ms exposure at 578 nm in the brown skin of Koreans. The energy density for clinical minimal whitening was 6-8 J/cm2, and the maximum penetration depth of these energy densities was 0.4 mm. We also found that the epidermal damage increased with increasing pulse widths at a fixed energy density (10 or 8 J/cm2) while the severity and depth of vascular damage decreased. These findings suggest that it is best to treat PWS with a copper vapor laser at the minimal pulse width and maximal power output possible at given energy density. CONCLUSION: We have demonstrated that the copper vapor laser treatment of PWS in the brown skin is not as selective as in white skin because of epidermal melanin. PMID- 8732575 TI - Electron paramagnetic resonance spectroscopy of free radicals in corneal tissue following excimer laser irradiation. AB - BACKGROUND AND OBJECTIVES: Free radicals, detected previously in corneal tissue following 193 nm laser irradiation, may be important agents in the laser/tissue interaction. Electron paramagnetic resonance spectroscopy (EPR) has been used to examine such radical formation in detail. STUDY DESIGN/MATERIALS AND METHODS: Bovine corneal strips were frozen in liquid nitrogen, irradiated with excimer laser pulses, and assayed by EPR. Exposure conditions were varied to study radical formation dependence on laser intensity and repetition. Results were measured against a quantifiable standard to calculate radical quantum yield. RESULTS: Either weak or intense laser fluences produced comparable tissue EPR signals. Radicals accumulated in frozen tissue for at least 10 initial ablation pulses. Radical quantum yield in cornea was 0.15%. CONCLUSION: Corneal radical formation is largely a photochemical process driven by the 193 nm laser radiation. Reactive radical species are produced in substantial numbers and likely have a significant clinical role. PMID- 8732576 TI - Study of corneal ablation with picosecond laser pulses at 211 nm and 263 nm. AB - BACKGROUND AND OBJECTIVE: Corneal ablation has been studied by picosecond laser pulses in the far-UV region. STUDY DESIGN/MATERIALS AND METHODS: Laser pulses of 25 ps duration at 211 nm and 263 nm wavelengths and a 1 kHz repetition rate have been used to ablate human and rabbit corneas. The dependence of the etch rate on laser fluence has been measured at both wavelengths. The collateral tissue damage has been investigated by electron microscopy. RESULTS: The ablation threshold for human cornea is determined to be about 3.0 mJ/cm2 at 211 nm, while the thresholds for rabbit cornea are about 2.3 mJ/cm2 at 211 nm and 8.0 mJ/cm2 at 263 nm. The slopes of the ablation curves and the dimensions of the damage zones have also been determined. CONCLUSION: We compare these results to the existing data on corneal ablation by nanosecond UV pulses and discuss the deficiency of the photochemical model. Experimental results are analyzed in terms of a model that features plasma ablation assisted by chromophore absorption. PMID- 8732577 TI - Excimer laser induced bubble: dimensions, theory, and implications for laser angioplasty. AB - BACKGROUND AND OBJECTIVE: Previous studies have demonstrated that during Xenon Chloride excimer laser ablation of tissue, rapidly expanding and imploding bubbles (diameter < 3 mm), predominantly containing water vapor, are formed. These short lived bubbles (life time < 300 microseconds) induce mechanical damage in adjacent tissue. In the present study, a theoretical analysis of the volume of vaporized water is correlated with measured bubble volumes formed in hemoglobin solution. STUDY DESIGN/MATERIALS AND METHODS: The dimensions of the rapidly expanding and imploding vapor bubble induced by the XeCl excimer laser pulses (308 nm, 115 ns), delivered via a 300, 550, or 950 microns diameter monofiber in 16% w/v hemoglobin solution (at 37 degrees C), were measured. RESULTS: Theoretical analysis and the experimental data correlated well (correlation coefficient r = 0.97). The diameter of excimer laser induced bubbles increased with increasing pulse energy. For a given radiant exposure, the bubble size was decreased by either decreasing the fiber tip area or by decreasing the absorption coefficient of the hemoglobin solution. CONCLUSION: We conclude that, for a wide range of conditions, theory agrees well with experimental data. Thus, during delivery of excimer laser pulses in blood, bubble dimensions can be reduced by flushing with saline or by reduction of the area radiated with each laser pulse, for example, by pulse multiplexing or using a smaller multifiber catheter. PMID- 8732578 TI - Copper vapor laser and photocarcinogenesis in hairless mice. AB - BACKGROUND AND OBJECTIVE: Port wine stains are commonly located on UV-exposed skin areas. We therefore examined the long-term interaction between UV radiation and copper vapor laser light (578 nm, yellow light) and whether the thermal influence from laser light had a carcinogenic potential itself. STUDY DESIGN/MATERIALS AND METHODS: The study was conducted in lightly pigmented hairless hr/hr C3H/Tif mice and included 8 groups of 17-20 mice. Intensities of 0.5, 1.0, and 1.4 W were used, corresponding to calibrated Hexascan fluences of 15.9, 31.8, and 44.6 J/cm2. Beam diameter was 1 mm and pulse duration 250 msec. UV irradiation of the mice was performed 4 days weekly and started the day after laser treatment. The UV simulated solar ultraviolet radiation can from one Phillips TL 12 and five Bellarium-S SA-1-12 tubes. The daily dose was 1.3 J/cm2, equivalent to 2.1 B-MED. RESULTS: No tumors appeared in groups receiving laser light only. The time to first (P < 0.01), second (P < 0.01), and third (P < 0.02) tumor was significantly delayed in the group treated with 1.4 W before UV irradiation (P < 0.01) compared with those receiving UV radiation only. No significant differences could be demonstrated for the groups treated with 0.5 and 1.0 W. CONCLUSION: One laser treatment with the copper vapor laser did not accelerate UV-induced photocarcinogenesis, and the laser exposure did not have a malignant potential itself. PMID- 8732579 TI - Effect of photodynamic therapy on the healing of a rat skin flap and its implication for head and neck reconstructive surgery. AB - BACKGROUND AND OBJECTIVE: Photodynamic therapy (PDT) may as adjuvant therapy by used to reduce tumor recurrence in the head and neck with surgery, given intraoperatively after resection. A concern with the use of intraoperative PDT is the possible effect on wound healing, especially on the healing of myocutaneous skin flaps, which are widely used to reconstruct defects following resections for head and neck cancer. STUDY DESIGN/ MATERIALS AND METHODS: A flap, based on the inferior epigastric artery, was prepared in thirty male Lewis rats. Group I did not receive any further treatment but the wound was left open for 20 minutes. Group II was injected with 5mg/kg Photofrin, 48 hours prior to the operation and also did not receive any further treatment. The wound bed and wound borders of group III were treated with 630nm light of different dosages, delivered by an argon dye laser. Animals in group IV received 5mg/kg Photofrin 48 hours prior to the operation and their wound beds were treated with the same light dosages as group III. After the treatment all flaps were replaced into the wound bed and the incisions were closed. Biopsies for histological analysis were taken at several time points; and on day 21, biopsies for wound tensile strength measurements were taken. RESULTS: The wound healing in group I, II, and III appeared normal and there were no differences seen between these groups. Also, the tensile strength did not differ significantly. The flaps of group IV showed serous effusion, epidermal necrosis, and weaker tensile strength (P = .04 and .02 for the light doses of 50 J/sq cm and 75 J/sq cm respectively) at a specific time point. CONCLUSION: The results of this study demonstrate that PDT given immediately before flap reconstruction will result in delayed wound healing. These results should be considered when contemplating the use of PDT as adjuvant intraoperative therapy for tumor surgery requiring flap reconstruction after ablative surgery. PMID- 8732580 TI - Pharmacokinetics of pyropheophorbide-a-hexyl ether in the dog. AB - BACKGROUND AND OBJECTIVES: Pyropheophorbide-a-hexyl ether (HPPH) is a new compound being investigated for use as a photosensitizer for photodynamic therapy; however, the pharmacokinetics are not known for any of the target species likely to be treated with this drug. The objective of this study was to determine the pharmacokinetic parameters of this drug prior to institution of a clinical trial in canine patients with various cancers. STUDY DESIGN, MATERIALS AND METHODS: HPPH (0.3mg/kg i.v.) was administered to 12 dogs and blood samples were drawn at intervals for 24 hours and plasma HPPH concentrations were determined. Pharmacokinetic parameters were calculated for each dog. RESULTS: No evidence of toxicity was noted in any dog. The mean half-life was calculated to be 26.98 +/- 2.35 hrs. The mean clearance was 5.061 +/- 0.214 ml/hr/kg. The mean volume of distribution of the central compartment was 0.069 +/- 0.003 L/kg, and the mean steady state volume of distribution was 4.47 +/- 0.25 L/kg. CONCLUSION: The conclusion is that 0.3 mg/kg HPPH injected intravenously resulted in measurable plasma levels for 24 hrs, and resulted in no detectable adverse reactions. PMID- 8732581 TI - Dynamic T1-weighted magnetic resonance imaging of interstitial laser photocoagulation in the liver: observations on in vivo temperature sensitivity. AB - BACKGROUND AND OBJECTIVE: Magnetic resonance imaging's (MRI) potential to monitor interstitial laser photocoagulation (ILP) has been previously demonstrated and is further tested here with improved spatial and temporal resolution. STUDY DESIGN/MATERIALS AND METHODS: In vivo experiments employed fiber-delivered 1,064 nm light (3.0 W, 150 sec) in six rabbit livers monitored under T1-weighted FSE MRI as 1 image/10 sec and a 3 mm thick 8 cm FOV. Image signal intensities (SI) were compared with temperatures (T) at 7, 10, and 15 mm from the fiber. RESULTS: Data showed 33 degrees C < T < 60 degrees C. SI did not vary inverselinearly with T; changes in the tissue altered the MRI signal interfering with the SI changes due to temperature. CONCLUSION: MRI cannot map SI-derived temperatures over the entire treatment site. The role of MRI's temperature sensitivity must be coordinated with organ- and dose-specific tissue changes. PMID- 8732582 TI - Sensitivity to Nd:YAG induced laserthermia is a cell-type-specific feature not directly related to tumorigenic potential or proliferation rate. AB - BACKGROUND AND OBJECTIVE: Laser-induced hyperthermia, laserthermia, is a promising new method for treating neoplasms. The response of different cell types to conventional hyperthermia varies [Bhuyan, Cancer Res 1979; 39:2277-2284; Raaphorst et al., Cancer Res 1979; 39:396-401]. We investigated the possible relationship between sensitivity to laser treatment and tumorigenic potential of three closely related cell types. Non-tumorigenic cells PYS-2 and differentiated F9S1 were compared to tumorigenic cells F9S1. STUDY DESIGN/MATERIALS AND METHODS: The contact Nd:YAG laser was used in a continuous-wave mode with a power setting of 6W, exposure times were 2 and 4 min [Castren-Persons et al., Lasers Surg Med 1991; 11:595-600; Castren-Persons, unpublished data]. The frosted-end probe was placed in the middle of the well and a thermocouple was attached 5 mm from the tip of the probe. The total amount of energy was measured for each well. A 4 min, 44 degrees C water bath treatment was used as comparison. Untreated wells served as controls. May-Grunwald-Giemsa staining and 3H-thymidine labeling were used for the analysis. RESULTS: Laserthermia killed all three cell types significantly more effectively than the water bath. PYS-2 cells were the most sensitive to the laser treatment. At the same temperature, PYS-2 cells were only slightly affected by water bath induced heating; the differentiated F9 cells were the most sensitive to this treatment. During the laser treatments, the energy required for holding the temperature seemed to depend not only on the cell type but also on the amount of cells treated: the more cells in the well, the more energy was needed. CONCLUSION: Our results suggest that laser sensitivity is a cell-type specific feature which is not directly related to the proliferation rate or benign or malignant behavior of the cells. PMID- 8732583 TI - Gender differences in recalled parental childrearing behaviors and adult self esteem. AB - A retrospective questionnaire study investigated gender differences in the relations between the self-reported self-esteem of 155 psychiatric adult outpatients and their recalled experience of their parents' behavior toward them as children. There were no significant sex differences in degree of self-esteem. However, it had a higher correlation to parenting variables for the men than for the women, with maternal predictor variables accounting for 36% and paternal predictors accounting for 32% of the variance in the men's self-esteem. Neither combined maternal nor combined paternal variables were significant predictors for women. For men, parental acceptance/ autonomy was significantly and positively related and inconsistency negatively related to self-esteem. Paternal rejection but not maternal rejection was significantly associated with low self-esteem only for the women. The greater amount of variance explained by childrearing variables in the men's self-esteem scores was attributed to the earlier ego development and consequent increased individuation in women. PMID- 8732584 TI - Is early separation anxiety a risk factor for adult panic disorder?: a critical review. AB - Heightened levels of early separation anxiety (SA) have long been linked to the risk of adult panic disorder (PD), suggesting that the two types of anxiety arise from a common diathesis--a proposition that has considerably influenced the classification of the anxiety disorders. However, the SA-PD link remains contentious, with some recent studies failing to confirm that putative association. All published research studies investigating the relationship of early SA to PD and/or other anxiety disorders were reviewed. Taken as a whole, the evidence provides support for the SA-PD hypothesis, although the specificity of that relationship needs further clarification. Problems of sample selection, retrospective measurement of early SA and comorbid diagnoses limit the certainty with which inferences can be drawn from existing data. Nevertheless, a recent community-based study provides additional support for the SA-PD hypothesis. Possible developmental pathways linking SA to PD are considered. One possibility that has not received adequate research attention is that early SA disorder (SAD) may persist into adulthood, rendering the sufferer vulnerable to panic and other anxiety symptoms when confronted with salient life stressors. We conclude that it is premature to reject the SA hypothesis of PD. Only well-designed longitudinal studies can map the complex developmental pathways linking early and later manifestations of morbid anxiety. PMID- 8732585 TI - ICD-10 checklists--a tool for clinicians' use of the ICD-10 classification of mental and behavioral disorders. AB - The Tenth Revision of the International Statistical Classification of Diseases and Related Health Problems (ICD-10) checklists are a set of semistructured instruments designed for clinician assessment of mental and behavioral disorders according to ICD-10 criteria. The checklists have been produced as a package consisting of the ICD-10 Symptom Checklist and Glossary and the International Diagnostic Checklists (IDCL) for ICD-10 and Manual. This report describes the main features of the newly released World Health Organization (WHO) instruments and specifies areas of possible application by clinicians. PMID- 8732586 TI - Schizophrenia and Alzheimer's disease: clinical and pathophysiologic analogies. AB - Psychotic symptoms are prominent in schizophrenia and a frequent neuropsychiatric manifestation of Alzheimer's disease (AD), occurring in approximately 50% of patients affected. The shared psychiatric symptoms suggest common cerebral pathophysiologies. Radiologic and pathologic findings indicate a predilection toward limbic involvement, with structural and atrophic changes of the medical temporal region predominating in both disorders. Neurochemical alterations affecting the dopaminergic/cholinergic axis appear to be central to both schizophrenia and AD. The basic pathologies of the two disorders are different, but they have similarities in the pattern of regional brain dysfunction, biochemical dysfunction, and symptomatology. We represent a selective review of these similarities. Insights drawn from these observations enrich the understanding of each disorder. PMID- 8732587 TI - Psychopathology: an edgeless razor? AB - The declining value of schneiderian first-rank symptoms (FRS) in differentiating between the two major functional psychoses has raised questions as to the utility of psychopathological concepts. In current diagnostic systems such as DSMs, psychopathological criteria and chiefly schneiderian symptoms have been used for nosographical purposes. It must be clear that this nosographically oriented use of psychopathology is only one aspect of the psychopathological enterprise. Indeed, while clinical psychopathology is essentially aimed at the identification of symptoms that are significant in view of nosographical distinctions, the specificendeavor of general psychopathology might be conceived as the organization of internal experiences around a core of meaningfulness, regardless of nosographical attributions. We seek to legitimate the value of psychopathological investigation and concepts as independent from any nosographical concern, and propose the concept of "psychopathological organizers" as synthesizing schemes of comprehension aimed at connecting different pathological experiences into unitary cores of meaningfulness. PMID- 8732588 TI - Dimensions of the Brief Psychiatric Rating Scale: an examination of stability during haloperidol treatment. AB - The purpose of the study was to investigate invariance of the factor structure of the Brief Psychiatric Rating Scale (BPRS) during haloperidol treatment. Data were collected in a placebo-controlled, double-blind, crossover treatment study examining the relationship between haloperidol plasma level and clinical outcome. Subjects were 173 acutely exacerbated, newly admitted patients with a diagnosis of schizophrenia or schizoaffective disorder. Exploratory (EFA) and confirmatory (CFA) factor analyses were performed at four time points: (1) last day of the preplacebo period; (2) last day of the placebo period; (3) end of the first treatment week; and (4) end of the treatment period. EFAs demonstrated (1) good reproducibility of commonly used BPRS factors at the preplacebo time point, (2) substantial changes in the factor structure during the placebo period, and (3) a reemergence of the initial (preplacebo) factor structure for most of the factors during haloperidol treatment. CFAs showed that traditional clinical factors were correlated with each other at all time points and that there were substantial changes in correlation among these factors over time. Our results suggest that longitudinal applications of traditional BPRS subscales should take factorial stability over time into consideration. Since many clinical trials use placebo data as a standard for longitudinal comparisons, further psychometric investigation is necessary to increase construct reliability and temporal stability. PMID- 8732589 TI - Patient dynamics in early stages of vocational rehabilitation: a pilot study. AB - Since little is known of the dynamics of the rehabilitation process, this pilot study aims to explore (1) the work performance and personal and social functioning of patients attending a vocational reintegration program; (2) any significant differences in course between successful and unsuccessful patients; and (3) whether these subgroups differed at entrance assessment. Using the 30 item Nurses' Observation Scale for inpatient Evaluation (NOSIE) and a global work performance scale, the sample (N = 31) showed a significant decrease in work performance and a negative trend in most NOSIE scores, instead of the expected steady progress. The turning point is reached after 9 to 12 weeks. In the entrance assessment, the failure subgroup (n = 11) displayed more negative and general symptoms and fewer social skills. It seemed to be not their initial work performance but their resources to cope with the stress of the program that were insufficient. However, not every downhill trend leads automatically to failure, and some patients still have a chance of later improvement. PMID- 8732590 TI - Suicide in the United States, 1951-1988: constant age-period-cohort rates in 40- to 44-year-old men. AB - Age-specific suicide rates in the United States between 1951 and 1988 were shown in cohort, cross-sectional, and age group formats. Despite considerable variation among women and younger and older men, suicide rates among 40- to 44-year-old men have remained stable. Since suicide is strongly influenced by psychosocial stressors, the constant suicide rates in middle-aged men suggest a consistency in the stress experienced by this population subgroup. PMID- 8732591 TI - Risk factors for early, cumulative, and fatal infections after heart transplantation: a multiinstitutional study. AB - BACKGROUND AND METHODS: By multivariable analysis, risk factors were identified for initial infection of any type, cumulative infections during the first 6 months and fatal infection among 2210 heart transplant recipients at 30 institutions. RESULTS AND CONCLUSIONS: Of the 1218 infections in 695 patients, bacterial infections were most frequent (47%), followed by viral (42%), fungal (8%), and protozoal (4%). Risk factors for earlier infection included older recipient age (p < 0.0001), ventilator support at time of transplant (p < 0.0001), ventricular assist device at time of transplant (p = 0.02), OKT3 induction therapy (p < 0.0001), donor black race (p = 0.0007), and positive donor cytomegalovirus serology (for cytomegalovirus infection) (p = 0.0007). Cumulative infections during the first 6 months were increased by older recipient age (p < 0.0001), ventilator support at transplant (p = 0.0004), ventricular assist at transplant (p = 0.009), Black donor (p = 0.03), female donor (p = 0.03), and OKT3 induction therapy (p = 0.005). The actuarial freedom from fatal infection was 96% at 1 year and 95% at 3 years. Risk factors for death from infection included very old (p = 0.002) and very young recipients (p = 0.004), ventilator support at time of transplant (p = 0.004), older donor (p < 0.0001), and longer donor ischemic time (p = 0.02). The risk of death from infection within the first 3 months exceeded 20% among older recipients (> 55 years) on ventilator support at time of transplantation who received an older (> 50 years) donor heart. PMID- 8732592 TI - Role of antibody and indirect antigen presentation in transplant-associated coronary artery vasculopathy. PMID- 8732593 TI - Prospective study of the circadian pattern of blood pressure after heart transplantation. AB - BACKGROUND: Previous reports indicate that heart transplant recipients lack a normal nocturnal decline in blood pressure. This prospective study was designed to determine the evolution of circadian blood pressure patterns after heart transplantation. METHODS: Twenty-four-hour ambulatory blood pressure and heart rate was measured in eight heart transplant recipients early (47 +/- 35 days) and late (740 +/- 10 days) after transplantation. RESULTS: Early transplant recordings and the normal control group recordings showed similar daytime systolic blood pressure but had different nighttime systolic blood pressure (138 +/- 15 mm Hg versus 112 +/- 9 mm Hg, p = 0.0002). The percent nocturnal change in systolic blood pressure showed a nocturnal increase in blood pressure in the early recordings versus a decrease in the healthy subjects (+4 +/- 2.7 versus -13 +/- 5.4, p < 0.0001). The late recordings showed a significant decrease in the nighttime systolic blood pressure (138 +/- 15 mm Hg versus 119 +/- 7 mm Hg, p = 0.011). The percent nocturnal change in systolic blood pressure was also significantly different between the early and late recordings (+4 +/- 2.7 versus 9 +/- 9, p = 0.0082) indicating a return of a nocturnal decline in systolic blood pressure. Similar patterns in diastolic blood pressure were observed. No significant change in the percent nocturnal change in heart rate occurred (-10 +/ 4.1 versus -7 +/- 5.5). CONCLUSIONS: Prospective follow-up of this heart transplant population showed that diurnal blood pressure variation is restored in some patients; diurnal variation is not related to corticosteroids, cyclosporine, or heart rate. PMID- 8732594 TI - Assessment of right and left atrial function in patients with transplanted hearts with the use of magnetic resonance imaging. AB - BACKGROUND: The purpose of this study was to determine the volumes and cyclic volume changes of the cardiac atria after heart transplantation in physically healthy recipients. METHODS: Ten heart transplant recipients (New York Heart Association grade I) entered the protocol. All operations had been made with classic biatrial anastomoses. Eight healthy volunteers were studied as controls. To measure the volumes of both atria and left ventricular cavity, we used a gradient-echo cine sequence. The 10 mm slices covered both atria over the total cardiac cycle. Images were planimetered, and simultaneous volumes were totalled to give the cavity volume at each time phase. RESULTS: The heart rate of the heart transplant recipients was faster than in controls, which resulted in higher cardiac output. The atrial minimum volumes were significantly larger, and fractional emptying was smaller in transplanted hearts when compared with controls. Reservoir and stroke volumes were smaller and conduit volumes were larger in transplanted hearts than in controls. The atrial filling and emptying rates were lower in heart transplant recipients than in controls. CONCLUSIONS: The classic surgical technique used in heart transplantation results in large atria with decreased volume changes and filling rates even in physically healthy transplant recipients. Magnetic resonance imaging is a reliable tool in assessment of atrial volumes and cyclic function after heart transplantation. PMID- 8732595 TI - Recipient-donor atrial synchronization benefits acute hemodynamics after orthotopic heart transplantation. PMID- 8732596 TI - Actuarial survival of heart-lung and bilateral sequential lung transplant recipients with obliterative bronchiolitis. AB - BACKGROUND: Obliterative bronchiolitis is a progressive form of obstructive airway disease that threatens long-term survival in lung transplant recipients. Its incidence and the long-term survival of lung transplant recipients with obliterative bronchiolitis are unknown. METHODS: The results of 89 heart-lung and 13 bilateral sequential lung transplant survivors beyond 90 days of their operation were analyzed. The date of diagnosis for obliterative bronchiolitis was established histologically (presence of submucosal fibrosis) or physiologically by a persistent reduction in the forced vital capacity to less than 0.7 for greater than 6 weeks. There were 43 patients without obliterative bronchiolitis and 59 patients with obliterative bronchiolitis. RESULTS: No differences were found in the mean age and gender ratios between the two groups. The actuarial 1-, 5-, and 10-year percentage freedom from obliterative bronchiolitis was 72 +/- 4.6, 30 +/- 5.6, and 15 +/- 7.4, respectively, with a median onset of 689 days (range 55 to 3404 days). About half the patients with biopsy-proven obliterative bronchiolitis had a fall in their forced expiratory flow at 50% of forced vital capacity/forced vital capacity nearly 4 months before fulfilling the forced expiratory volume in 1 second criteria established by the Working Group on chronic lung dysfunction. The actuarial 1-, 5-, and 10-year percentage survival of obliterative bronchiolitis negative patients was 90 +/- 4.5, 74 +/- 8.4, and 66 +/- 10.6, respectively, versus 90 +/- 3.9, 49 +/- 6.9, and 27 +/- 10.0, respectively, for obliterative bronchiolitis positive patients (p = 0.38). The actuarial 1-, 3-, 5-, 8-, and 10-year percentage survival of lung transplant recipients after the diagnosis of obliterative bronchiolitis was 74 +/- 5.8, 50 +/- 7.5, 43 +/- 7.8, 23 +/- 8.7, and 11 +/- 9.1, respectively, with a median survival of 1084 days (range 0 to 3442 days). CONCLUSIONS: The forced expiratory flow at 50% of forced vital capacity/forced vital capacity is a more sensitive indicator for the early detection of obliterative bronchiolitis than the forced expiratory volume in 1 second after heart-lung or bilateral sequential lung transplantation. The obliterative bronchiolitis negative group survival tends to be better than the obliterative bronchiolitis positive group. The obliterative bronchiolitis positive lung transplant recipients have reasonable outcomes with a median survival time of nearly 3 years after the diagnosis of obliterative bronchiolitis. Earlier detection of obliterative bronchiolitis and refinements in management may further improve these results. PMID- 8732597 TI - Yield of surveillance transbronchial biopsies performed beyond two years after lung transplantation. AB - BACKGROUND: Acute lung rejection after transplantation may lead to significant graft dysfunction. Hence surveillance protocols which include transbronchial biopsies have been established to monitor for early findings suggestive of acute rejection. However, the frequency of acute rejection diminishes as time from the transplant procedure increases. We therefore sought to examine the incidence of acute rejection seen on surveillance transbronchial biopsy performed at least 2 years after transplantation. METHODS: A retrospective review of all transbronchial biopsy was conducted. Forced expiratory volume in 1 second at the time of transbronchial biopsy and in the preceding 6 months was recorded. RESULTS: A total of 102 transbronchial biopsy procedures were assessed. Histologic evidence of at least grade 2 rejection (including three judged as grade 1 to 2) occurred in biopsy samples from 10 procedures (9.8%). Five of the episodes occurred at 2 years, three occurred at 3 years, and two occurred at 4 years. Inadequate biopsies occurred on 22 occasions (21.6%). A corresponding significant change in forced expiratory volume in 1 second ( >or= 10% decline) compared with the preceding 6 months was seen in 4 of 10 episodes of grade 2 acute rejection. Only one of five episodes occurring at least 3 years after transplantation was associated with a decline in forced expiratory volume in 1 second. No evidence of acute rejection was found in any of 15 transbronchial biopsy procedures beyond 4 years after transplantation where adequate material for histologic interpretation was obtained. However, no statistically significant change was found in the proportion of transbronchial biopsy procedures showing acute rejection as a function of time. CONCLUSIONS: We conclude that transbronchial biopsy-proven acute rejection may occur in patients with asymptomatic, stable conditions who have survived at least 2 years, but the yield of transbronchial biopsy performed beyond 4 years is exceedingly low. PMID- 8732598 TI - Single lung transplantation followed by contralateral bullectomy for bullous emphysema. AB - BACKGROUND AND METHODS: Single lung transplantation for emphysema is now standard practice despite initial concerns, including the possibility that the compliant diseased lung would compress the transplanted lung as a result of hyperinflation. We describe a patient with severe bilateral bullous emphysema and alpha-1 antitrypsin deficiency who underwent single lung transplantation after which hyperinflation of the native lung led to significant compression of the pulmonary allograft. The patient subsequently underwent bullectomy of the contralateral lung with marked improvement in his functional status. RESULTS: After bullectomy, the patient's forced expiratory volume in 1 second increased from 1.77 to 2.82 L, his total lung capacity fell from 7.23 to 6.19 L, and his 6-minute walk increased from 724 to 1269 feet. However, 7 months after bullectomy, there was evidence that the bullous disease in the native lung was recurring. CONCLUSIONS: Significant hyperinflation of the native lung with compromise of the pulmonary allograft can occur after single lung transplantation for bullous emphysema. Bullectomy of the diseased lung after transplantation improved allograft function in our patient. Alternatively, bilateral lung transplantation for severe bilateral bullous emphysema may be considered. PMID- 8732599 TI - Retrograde versus antegrade flush in canine left lung preservation for six hours. AB - BACKGROUND: Retrograde flush through the left atrium is now used by some investigators in clinical lung preservation. However, to date there are no studies which compare its result with that of routine antegrade flush. METHODS: Mongrel dogs were divided into two groups: antegrade group (n = 7) and retrograde group (n = 8). After flush and 6 hours of cold storage in Euro-Collins solution, the left lung was transplanted in weight matched recipients, and their right pulmonary artery was then clamped at 10-, 30-, 60-, and 120-minute intervals for 10 minutes to test the lung function. The ultrastructure of lungs in both groups were also studied. RESULTS: Results showed the following (antegrade group versus retrograde group): the wet/dry ratio of the transplanted lung was 7.14 +/- 0.15 versus 6.33 +/- 0.20 (p < 0.01); the arterial oxygen tension (mm Hg) was 389 +/- 42 versus 534 +/- 23 (p < 0.05) and 370 +/- 51 versus 580 +/- 37 (p < 0.01) at 60 and 120 minutes, respectively. The peak airway pressure (cm H2O) was 23.4 +/- 0.8 versus 20.6 +/- 0.6 (p < 0.05) and 23.7 +/- 0.6 versus 21.3 +/- 0.8 (p < 0.05) at 10 and 60 minutes, respectively. Electron microscopic studies showed that at the end of preservation, type I and type II pneumocytes and capillaries were normal in both groups. Occluded capillaries with red blood cells were found in the antegrade group. After reperfusion, damaged epithelium and thicker air-blood barrier were found in the antegrade group. CONCLUSIONS: Retrograde flush offers a better lung preservation with less edema, decreased airway resistance, and improved oxygenation as compared with the antegrade group in 6 hours lung preservation. PMID- 8732600 TI - Receptor-specific effects of acute rejection after heart transplantation on endothelium-dependent coronary relaxation in dogs. AB - To establish the effects of acute rejection on coronary artery endothelial reactivity in a canine model of heart transplantation, we submitted seven mongrel dogs to heterotopic heart transplantation without immunosuppression for a period of 7 days. At harvesting, all transplanted hearts displayed a grade IV histologic rejection. Compared with native heart, the endothelium-dependent relaxation of the coronary arteries from the graft displayed an increased sensitivity to serotonin, a decreased sensitivity to thrombin, and no change in the response to acetylcholine and adenosine diphosphate; endothelium-independent relaxation to sodium nitroprusside was not affected. Therefore, in the canine heterotopic heart transplant model, acute rejection has no effect on endothelium-independent relaxation of coronary arteries but affects endothelium-dependent relaxation in a receptor-specific manner. PMID- 8732601 TI - Thromboxane receptor blockade does not attenuate pulmonary pressor response in porcine single lung transplantation. AB - BACKGROUND: The ischemia-reperfusion lung injury is characterized by increased pulmonary vascular resistance, edema, and subsequent deterioration of oxygenation. Other models of acute lung injury suggest that thromboxane A2 may contribute to the pulmonary hypertension after transplantation. METHODS: We studied the effects of the selective thromboxane A2 receptor antagonist SQ 30741 on pulmonary hemodynamics and gas exchange in porcine single lung transplantation using extracorporeal circulation (right heart bypass) with separate cannulations of the right and left pulmonary arteries. Pulmonary vascular resistance was measured at equal and constant flow to each lung. Flow distribution between the lungs was registered at equal pulmonary artery pressures. Twelve pigs (weight 17 to 23 kg) were studied. At the onset of reperfusion a bolus dose of the drug (5 mg/kg) was injected into both pulmonary arteries followed by an infusion (5 mg/kg/hr) for 1 hour (SQ group, n = 6). The control group (n = 6) received an equal amount of vehicle. The systemic and pulmonary hemodynamics and blood gas values were registered during 2 hours of reperfusion. RESULTS: The pulmonary vascular resistance of the transplanted lung was significantly higher compared with the native lung (p < 0.001). Administration of SQ 30741 failed to ameliorate the pulmonary pressor response of the graft in comparison with the control group. No difference was found in the systemic arterial oxygen tension between the two groups. CONCLUSIONS: Thromboxane does not seem to be among the principal mediators in the pulmonary hypertension after transplantation. PMID- 8732602 TI - Experience of FK506 immune suppression in pediatric heart transplantation: a study of long-term adverse effects. AB - BACKGROUND: Immunosuppression with FK506 for pediatric heart transplantation has been used in this institution since 1989. This study reports the unique toxicity of this macrolide agent in these heart transplant recipients. METHODS: Between October 1989 and August 1994, 49 patients were managed with FK506, which was the initial primary agent in 38 patients. The remaining 11 were switched from cyclosporine A because of persistent rejection or side effects from the cyclosporine A or prednisone. Data were obtained retrospectively from medical records. RESULTS: Mean duration of follow-up was 29 months (median 37 months, range 3 to 96 months). Twenty-nine patients (59%) were receiving FK506 alone; 20 patients (41%) were receiving additional treatment with azathioprine, prednisone, or methotrexate. There were seven deaths. Twenty patients (41%) had elevated creatinine levels between 1 to 2 mg/dl. Five patients (11%) had levels greater than 2 mg/dl. Two patients with preexisting renal dysfunction while receiving cyclosporine A had chronic renal failure 32 and 36 months after switching to FK506 and required kidney transplantation. Hyperkalemia was a persistent finding in 26 patients. Of eight patients with hypertension, four had preexisting disease while receiving cyclosporine A; two had impaired renal function, and two were receiving prednisone. Severe anemia developed in eight patients (16%), two of whom had parvovirus. Moderate anemia developed in 21 patients (43%). Eosinophilia occurred in 19 patients; 11 of 19 patients (58%) had allergic symptoms. There was one case of diabetes mellitus. There were 12 significant infections with four infection-related deaths. Lymphoproliferative disease was noted in three patients, two of whom survived. Gastrointestinal symptoms, including chronic diarrhea, recurrent abdominal pain, and reflux esophagitis were present in 10 patients. CONCLUSIONS: In our experience, anemia, renal toxicity, hyperkalemia, chronic diarrhea, and allergies were the most common adverse effects of FK506. Unlike cyclosporine A, hypertension, gingival hyperplasia, coarsening of facial features, and hirsutism were not seen. PMID- 8732603 TI - Early progressive mobilization of patients with left ventricular assist devices is safe and optimizes recovery before heart transplantation. AB - BACKGROUND: As early perioperative survival with left ventricular assist device support improves, additional emphasis must be placed on patient rehabilitation. Especially as mean left ventricular assist device support times exceed 100 days, it is essential for health professionals to mobilize patients to prevent the incidence of the deleterious effects of bedrest. The timing of optimal functional performance and the safety of rehabilitation has not been described. METHODS: We retrospectively studied patients surviving left ventricular assist device implantation (34 of 41 patients; 27 men, 7 women; age 51 +/- 11 years). Physical therapy consisted of progressive mobilization leading to treadmill exercise or cycling. All patients exercised in the "pump on full" mode with flows >or= 3.0 L/min. RESULTS: Twenty of thirty-four patients initiated ambulation at 7 to 10 days with independent ambulation by 14 days in 55% of the group. Treadmill exercise was tolerated by 82% of the patients, begun at postoperative day 21 by eighteen patients. The greatest improvement in exercise performance was seen by 6 to 8 weeks (20 to 30 minutes at 3.17 +/- 0.79 metabolic equivalents). Maximal functional capacity achieved was influenced by medical complications. A total of 1878 treatment sessions lasting 1390 hours was performed. Only four minor incidents occurred representing 2.9 incidents/1000 patient hours; all involved a transient decrease in pump flow. None of these events resulted in an increase in morbidity or mortality. CONCLUSIONS: Progressive mobilization in patients with left ventricular assist device is safe. Patients return to independence in activities of daily living and tolerate prolonged workloads of up to 5 metabolic equivalents. There is rapid improvement in functional capacity until 6 weeks after operation. Delay in transplantation until this time may optimize postoperative recovery. PMID- 8732604 TI - Twenty-three-year survival after orthotopic heart transplantation. PMID- 8732605 TI - Left atrial dysfunction after orthotopic heart transplantation. PMID- 8732606 TI - Pain and stress in infancy and childhood--where to now? PMID- 8732607 TI - Acute respiratory distress syndrome (ARDS) in neonates and children. AB - ARDS remains a syndrome which despite all efforts poses problems in exact definition (cause, course and severity). Most of the existing information comes from clinical observations and uncontrolled studies and is therefore of limited value. Despite the advent of new treatment modalities mortality from ARDS has remained high and is influenced or caused by several factors like underlying disease, previous health status, presence of MOSF, complications of therapy or ultimate failure of gas exchange. Therapy is directed at elimination of the cause of ARDS if possible, but then mainly supportive, considering all organs and systems. With the introduction of gentler respiratory support techniques (small tidal volumes and pressure limitation, permissive hypercapnia and HFO) and appropriate measures to reduce oxygen toxicity (titration of PEEP, possibly NO), iatrogenic lung injury, indistinguishable from ARDS, can be reduced, and this might improve survival rates. For the future, modulation of the host's inflammatory response may hold great promises for prevention and treatment of ARDS, but such strategies need to be explored with well controlled clinical trials, respecting the complexity of the issue. PMID- 8732608 TI - Parents in the anaesthetic room: a questionnaire survey of departments of anaesthesia. AB - Thirty departments of paediatric anaesthesia in the United Kingdom were sent a questionnaire about their policies and practices regarding parental presence in the anaesthetic room. Of 22 respondents, 100% left the decision about whether a parent should be present for induction of anaesthesia to the individual anaesthetist and only 14% had written policy guidelines. All departments permitted parental presence at induction for elective surgery, compared with 77% for emergency surgery and 55% for a rapid sequence induction. The minimum age of child below which parents were not allowed at induction varied between no age limit and one year. PMID- 8732609 TI - Parental presence during induction of anaesthesia: practice differences between the United States and Great Britain. AB - A questionnaire was sent to 1353 paediatric anaesthetists in Great Britain and the United States. Nineteen questions were asked about attitudes toward parental presence during induction of anaesthesia and the prevalence of such practice. Overall, respondents from Great Britain support parental presence more than the United States respondents. For example, 82% of the Great Britain respondents, vs 64% of the United States respondents thought that parental presence during induction decreases the anxiety (P = 0.001) and increases the cooperation of the child (P = 0.001). Most United States respondents (58%) allow parental presence in less than 5% of their cases, but most Great Britain respondents (84%) allow parental presence in more than 75% of their cases. We conclude that in contrast to the respondents from Great Britain, the majority of the United States sample does not feel that parental presence is useful and so does not routinely use this technique in their practice. PMID- 8732610 TI - Spinal anaesthesia in newborns: total and free bupivacaine plasma concentration. AB - This study was designed to measure total and free bupivacaine (B) after spinal anaesthesia in newborns and to evaluate a possible influence of adrenaline on B absorption. Twenty-two newborns were randomly allocated to receive either plain B (group 1) or adrenaline added to B (group 2) for spinal anaesthesia. A single blood sample was collected ten min after spinal injection. Total B concentration was found 0.31 +/- 0.17 microgram ml-1 in group 1 and 0.25 +/- 0.09 microgram ml 1 in group 2. Bound B concentration was 0.27 +/- 0.17 microgram ml-1 in group 1 and 0.22 +/- 0.09 microgram ml-1 in group 2. No difference was found between the two groups for these parameters. Albumin but not alpha 1-acid-glyco-protein correlated to age and weight, bound B correlated to alpha 1-acid-glyco-protein but not to albumin. Despite the low plasma concentration of binding proteins in newborns, spinal anaesthesia with B does not result in a high level of free drug. Adrenaline does not have any pharmacological advantage in these patients. PMID- 8732611 TI - Behavioural changes and children's memories after intravenous, inhalation and rectal induction of anaesthesia. AB - Ninety two-to-seven-year-old children admitted for routine day case ENT operations were randomly allocated to have either intravenous thiopentone (group I), halothane inhalation (group II) or rectal methohexitone (group III) for anaesthesia induction. Using a postoperative questionnaire, the parents evaluated the changes in the child's behaviour one day, one week and one month after the operation. Problematic changes were detected in 17 (59%) children in group I, 14 (50%) in group II and 17 (58%) in group III (NS). Although the children in group II behaved most calmly during the induction they had significantly more negative memories of the induction of anaesthesia (six children in group II vs two in group I and one in group III) and of the hospital in general (17 in group II vs 11 in group I and eight in group III) than children in the other groups. Judging from memories of young children, intravenous and rectal inductions are less distressing to them than inhalational induction. PMID- 8732612 TI - Deep sedation with propofol in preschool children undergoing radiation therapy. AB - Immobilization of children undergoing radiation therapy always requires anaesthesia. Deep sedation with continuous infusion of propofol and spontaneous breathing, (we call it ?sedative anaesthesia'), may be an alternative to general anaesthesia with intubation and controlled ventilation. This clinical report deals with 155 anaesthetics performed in 11 consecutive paediatric oncology patients, mean age 30 months (range 19-42), who required radiation therapy for from seven to 33 consecutive days. Mean duration of anaesthesia was 18 ( +/- 11) mins. For induction, a loading dose of 3.6 (SD +/- 0.59) mg.kg-1 propofol was administered immediately followed by a continuous infusion of 7.4 ( +/- 2.2) mg.kg-1.h-1 for maintenance of anaesthesia. There were no complications of clinical importance involving respiration, circulation or neurology, except for one short episode of transient desaturation, which was managed by suctioning and changing head position. Children opened their eyes spontaneously four ( +/- 3.7) min after discontinuing the propofol infusion and could be discharged about 30 mins later. Tachyphylaxis or unpleasant side effects during and after anesthesia have not been observed. Sedative anaesthesia with propofol seems to be an excellent method to immobilize paediatric patients during radiotherapeutic procedures. PMID- 8732613 TI - Postoperative pain in children: a survey of parents' expectations and perceptions of their children's experiences. AB - Parental expectation and participation in postoperative analgesia is very important in paediatric practice. In order to improve postoperative pain management in children, the parents of 31 elective surgical children, three months to 15 years of age, were asked preoperatively about their expectations regarding their children's postoperative pain and pain relief. At 24 h after surgery, the parents were asked about their perceptions of their children's pain and pain control. The survey indicates that the parents had high expectations of good pain relief. They wanted effective analgesia administered promptly when the children had some pain. However, current practice in controlling pain after surgery is still not optimal. Nine (29%) of the children experienced severe or unbearable pain or experienced pain for the whole of the 24 h after surgery. An approach to improve pain management in children could be for the hospital staff to reorganize and to develop an 'acute pain service'. A pain service may not require new technology, but instead be based on more effective communication and skill in utilizing the traditional systems. PMID- 8732614 TI - Nasal mask positive pressure ventilation in paediatric patients with type II respiratory failure. AB - We report our experience with nasal mask ventilation in children and adolescents with type II respiratory failure admitted to the paediatric intensive care unit (PICU) over an 18-month period. Seven patients were treated with nasal mask ventilation during part of their PICU stay. All showed significant improvement in arterial pH, PaCO2, and PaO2/FiO2 from presentation to discharge, although at discharge PaCO2 and PaO2/FiO2 fell outside of the normal range. Complications occurred in four patients. When compared to 11 patients with type II respiratory failure not treated with nasal mask ventilation, the nasal mask ventilation group had a similar PICU length of stay and incidence of complications. We conclude that nasal mask ventilation may be useful in maintaining near normal alveolar ventilation in selected children with type II respiratory failure and that a prospective study of this technique is indicated. PMID- 8732615 TI - Craniodiaphyseal dysplasia; another cause of difficult intubation. AB - A nine-year-old boy with craniodiaphyseal dysplasia (CDD) presented for mandibular reduction. Patients with CDD present problems to the anaesthetist, specifically difficulties with airway management and tracheal intubation. This child was managed using laryngeal mask airway (LMA) guided fibreoptic intubation. Spontaneous respiration was maintained throughout intubation, following which ventilation was controlled and anaesthesia was provided using nitrous oxide, isoflurane and fentanyl. The perioperative management is described. PMID- 8732616 TI - Anaesthesia and the Beckwith-Wiedemann syndrome. AB - Infants with Beckwith-Wiedemann syndrome usually present different abnormalities which may require surgical correction. Anaesthetic management may be complicated by abnormal airway anatomy, congenital heart disease and severe hypoglycaemia. Careful preoperative evaluation, perioperative monitoring and suitable choice of anaesthetic technique are required for a successful outcome. We report the perioperative management of a patient with Beckwith-Wiedemann syndrome presenting for omphalocoele surgery on his first day of life and for bilateral inguinal hernia repair four months later. PMID- 8732617 TI - Anaesthetic management of the Kenny-Caffey syndrome using the laryngeal mask. AB - We report the anaesthetic management of a nine-year-old, 6.8 kg, 75 cm tall female with the Kenny-Caffey syndrome presenting for strabismus surgery. Dysmorphic features in our patient included a hypoplastic mandible. A neonatal (size 1) laryngeal mask was successfully used for management of the airway whilst providing surgical access. The general features of this rare syndrome are presented and the literature reviewed. PMID- 8732618 TI - Total spinal anaesthesia following caudal block with bupivacaine and buprenorphine. AB - This is a case report of an 18-month-old 10 kg child who presented for emergency repair of a recurrent diaphragmatic hernia with a history of craniofacial dystosis and was given a caudal block postoperatively with a combination of 4 ml of 0.5% bupivacaine and 2.5 micrograms.kg-1 buprenorphine made up to a total volume of 10 ml. An inadvertent dural puncture occurred resulting in total spinal block which was managed symptomatically. Block regression started one h later when the respiratory movements became noticeable. Eye opening and hand movements returned 3 h later. The patient's exposure to a large intrathecal dose of buprenorphine did not lead to prolonged respiratory depression. The possibility of a midbrain insult due to a sudden rise in intracranial pressure is also discussed. PMID- 8732619 TI - Blood loss from remote injection site. PMID- 8732620 TI - Propofol and green urine. PMID- 8732621 TI - Cannulation in an odd place. PMID- 8732622 TI - Leg blocks in children. PMID- 8732623 TI - Delayed recovery from anaesthesia in Rubinstein-Taybi syndrome. PMID- 8732624 TI - Antiphospholipid antibodies and pregnancy disorders in women with insulin dependent diabetes. AB - Insulin dependent diabetes (IDD) is considered to be an immune endocrinopathy as in such patients a disorder of the immune system is involved; however, up to now no data are available on the occurrence of antiphospholipid antibodies (aPL) in IDD pregnant women and on possible correlation between the presence of aPL and the high fetomaternal morbidity reported in these patients. The presence of lupus anticoagulant (LA) and of anticardiolipin antibodies (ACA) was monthly evaluated. In 35 IDD pregnant women referring within the 7 degrees week of pregnancy to the High Risk Pregnancy Medical Unit. Levels of D-dimer, fibrin degradation product, were also assayed. Twelve IDD pregnant women resulted to be aPL positive with a markedly high prevalence of positivity (34%). aPL positive did not significantly differ from aPL negative women in age, duration and severity of diabetes and in metabolic control throughout pregnancy. Pregnancy induced hypertension (PIH) and intrauterin growth retard (IUGR) were observed in 6/12 aPL positive and in only 2/23 aPL negative patients (p < 0.02). A pathological increase in D-dimer levels occurred in 6/12 aPL positive patients and in none aPL negative (p < 0.03). The high frequency of aPL positivity and its strict relation to pregnancy complications strongly support a major role for an autoimmune pathogenetic mechanism in the occurrence of feto-maternal morbidity in IDD pregnant women. The identification of this subgroup at risk for complications may be clinically relevant. PMID- 8732625 TI - A modification of the APC resistance test and its application to the study of patients on coumarin therapy. AB - APC resistance appears to be caused, predominantly, by a mutation in coagulation factor V (nucleotide 1691: G to A). This phenomenon is usually studied by performing APTTs in the absence and presence of added APC. We studied a modification of the assay involving dilution of the test plasma in factor V deficient plasma, to render the assay more factor V specific. This modification was applied to 76 patients with venous thrombosis on coumarin treatment and to 45 controls. Two out of 45 controls (4.4%) showed abnormal results with the modified test. They also showed loss of factor V exon 10 Mnl I restriction site, associated to APC resistance. All remaining controls, with normal functional results by the modified assay, showed normal restriction profile. We detected 9 affected patients (11.8%), one of them homozygous or double heterozygous. In conclusion, the modified assay is very sensitive for factor V dependent APC resistance, and can successfully be applied to patients on coumarin therapy. PMID- 8732626 TI - Activation of recombinant human protein C. AB - We have produced recombinant human Protein C (rHPC) in the milk of transgenic swine. After purification, we have analyzed the interaction of teh zymogen with Protac, thrombin/thrombomodulin and thrombin alone. The amidolytic and anticoagulant activities of rAPC after Protac activation were approximately 80% those of its human plasma counterpart. Upon the excision of the activation peptide by thrombin/thrombomodulin complex, both the natural and recombinant activation products had similar enzymatic and biological activities. This observation can be attributed to the difference in the mechanism of action between the two activators and structural differences between HPC and rHPC. PMID- 8732627 TI - Clinical significance of activated protein C resistance as a potential marker for hypercoagulable state. AB - The activated protein C (APC)-resistance test is a simple and reliable method for detecting reduced sensitivity to the anticoagulant action of this protein. We investigated the sensitivity to APC in 180 Japanese controls and in 96 Japanese patients with venous and arterial thrombosis (28 with deep vein thrombosis; 13 with pulmonary thromboembolism; 41 with cerebral infarction; and 14 with coronary artery disease). All of the patient groups showed significantly reduced sensitivity to APC, reflected by the lower normalized APC-sensitivity ratio (n APC-SR), as compared with healthy control. The APC-sensitivity ratio was negatively correlated with plasma activated factor VII levels. These results suggest that the low n-APC-SR is related to venous or arterial thrombotic disease. The APC resistance may serve as a potential marker for assessing the hypercoagulable state. PMID- 8732628 TI - The association of leukocyte count, fibrinogen and C-reactive protein with vascular risk factors and ischemic vascular diseases. AB - In 154 subjects (age 63 +/- 11 years; 63 women and 91 men) randomly selected from the population, we tested the hypothesis that inflammatory parameters are associated with vascular risk factors and particularly with a history of ischemic vascular diseases. The subjects were part of the control group (n = 197) in a case-control study investigating recent infection as a risk factor for acute cerebrovascular ischemia and had been matched for sex and age with patients suffering from acute ischemic stroke or transient ischemic attack. Subjects with malignant or inflammatory diseases, with recent trauma, surgery or vascular diseases (n = 43) were excluded from the present analysis. In multivariate analysis, current smoking, diabetes mellitus, age > or = 65 years, and a history of stroke independently increased the leukocyte count. Hypertriglyceridemia, peripheral arterial disease, and diabetes mellitus were positively associated with C -reactive protein (CRP). Age > or = 65 years and diabetes mellitus independently increased fibrinogen. (p < 0.05, respectively) Subjects with a history of cerebrovascular, cardiovascular or peripheral arterial disease had higher leukocyte counts, fibrinogen and CRP than subjects without vascular risk factors and higher leukocytes and fibrinogen than subjects with one or more risk factors. Subjects under the age of 65 with vascular risk factors but without ischemic diseases had higher leukocyte count, fibrinogen and CRP and subjects older than 65 with risk factors had higher CRP than subjects without risk factors or ischemic diseases in the same age group. (p < 0.05, respectively) These results support the hypotheses that low-grade inflammation is associated with vascular risk factors and that inflammatory mechanisms may contribute to the risk of organ ischemia. PMID- 8732629 TI - Flow cytometric evaluation of the effect of various thrombin inhibitors on platelet activation in whole blood. AB - In an in vitro study the effect of various thrombin inhibitors (argatroban, efegatran, DuP 714, recombinant hirudin and PEG-hirudin) on platelet activation in whole blood was investigated. Blood was drawn from normal human volunteers using the double syringe technique without use of a tourniquet to avoid autoaggregation of platelets. Blood was anticoagulated with either argatroban, efegatran, DuP 714, hirudin or PEG-hirudin at final concentrations of 10 micrograms/ml. Blood samples were then incubated at 37 degrees C either with saline, r-tissue factor, arachidonic acid, adenosine diphosphate or collagen. At definite times (1, 2.5, 5, 10 min) aliquots were taken and after various steps of fixative procedure the percentage of platelet activation was measured using fluorescent monoclonal antibodies to platelet surface receptors GPIIIa (CD-61) and P-selectin (CD-62). Flow cytometric analysis showed a platelet activation after all agonists used. All thrombin inhibitors studied caused a nearly complete inhibition of r-tissue factor-mediated platelet activation. In contrast, after activation with the other agonists an increased percent CD-62 expression was found with a maximum after 2.5 to 5 min. The results show that in whole blood thrombin inhibitors are effective in preventing platelet activation induced by r tissue factor. The formation of active serine proteases including thrombin may be effectively inhibited by these agents. The observations further suggest that while thrombin inhibitors may control serine proteases, these agents do not inhibit the activation of platelets mediated by other agonists. PMID- 8732630 TI - Quantification of kininogens in plasma. A functional method based on the cysteine proteinase inhibitor activity. AB - We have previously reported on a microassay based on human kininogens as cysteine proteinase inhibitors (CPIs), which could quantify partially purified kininogens from different biological fluids (J Pharmacol Meth 26, 113-124, 1991). In the present study we describe a functional method that, when assuming a 1:1 stoichiometry between papain and kininogen, allows a direct measurement of the molar concentration of kininogens in plasma. The principle of the method is that the target enzyme papain is inhibited by kininogens present in added diluted plasma. The residual activity of papain, not inhibited in this reaction, subsequently hydrolyzes the added peptide substrate (S-2302), generating a yellow colour which is read in a microplate reader at 405 nm. Relating the test samples to a standard curve established from known concentrations of E-64 (a selective low molecular weight inhibitor of cysteine proteinases), we could quantify kininogens on a molar basis. A major problem when first applying this method to plasma, was the interference of alpha 2-macroglobulin, which inhibited papain and generated a complex able to split the chromogenic substrate. The interference of alpha 2-macroglobulin was eliminated by an initial acid treatment of plasma followed by dilution with a buffer containing methylamine. The specificity for kininogens in this assay is demonstrated by the following observations: Commercial pooled normal plasma contained 3.2 microM CPI activity, in good agreement with the expected molar concentration of kininogens. After gel filtration of a plasma sample with a CPI activity of 3.4 microM, two peaks with CPI activity were identified as H-kininogen (0.9 microM) and L-kininogen (2.5 microM), both in good accordance with expected concentrations of the two kininogens. Plasma deficient of kininogens had a minimal inhibitory capacity towards papain. PMID- 8732631 TI - Analysis for antithrombin gene polymorphisms in Japanese subjects and cosegregation studies in families with hereditary antithrombin deficiency. PMID- 8732632 TI - Factor V Leiden-dependent APC resistance: improved sensitivity and specificity of the APC resistance test by plasma dilution in factor V-depleted plasma. PMID- 8732633 TI - Autologous stem cell transplantations. AB - High-dose treatments with autologous stem cell support have increasingly been used to improve the treatment results of a variety of haematological and nonhaematological malignancies. High-dose treatments cause severe bone marrow injury, which can effectively be rescued with infusion of a sufficient number of stem cells. Stem cells can be collected from bone marrow or by leukaphereses from blood. Before leukaphereses, stem cells--enumerated as CD34+ cells--must be mobilized from bone marrow to blood. The use of blood-derived stem cells for transplanting has certain advantages over bone marrow cells, one of the most important being the more rapid haemopoietic recovery from bone marrow ablation. As a result of the short cytopenic period, transplantation-related mortality is usually low. In this short review, the background of autotransplants, prerequisites for a successful blood cell transplant, clinical issues and future aspects are briefly discussed. PMID- 8732634 TI - Ten years after Chernobyl. AB - This review focuses on current findings on the health consequences of the accident at the Chernobyl nuclear power plant in 1986. Acute radiation damage caused by exposure to high doses of ionizing radiation involved a few hundred people, apparently with fewer than 100 deaths within the first few months. Epidemiologically detectable increases in congenital abnormalities have not been reported thus far, with the possible exception of Down's syndrome. Owing to the long latency period of radiation-induced cancer, discernible increases in cancer incidence and mortality are not yet expected for most tumour types, especially among adults. However, dramatic increases in the number of childhood thyroid cancers have already been observed in Belarus and Ukraine and the Bryansk regions of Russia. The increase has been over 100-fold in some areas with heavy contamination. From the viewpoint of overall public health, the outlook of direct health effects of the Chernobyl accident are likely to be severe only among some limited subgroups, such as young children exposed to high levels of fallout nuclides. In absolute terms, the global number of Chernobyl-associated cancer cases can be estimated to be tens of thousands, but only a small fraction of these is likely to be discernible epidemiologically. PMID- 8732635 TI - Gene therapy for cardiovascular diseases. AB - Gene therapy offers a promising alternative for the treatment of various cardiovascular diseases. Blood vessels are among the easiest targets for gene therapy and in some pathological conditions only a temporary expression of the transfected gene will be required to achieve a beneficial biological effect. These conditions include the prevention of postangioplasty restenosis, postbypass atherosclerosis, peripheral atherosclerotic vascular disease and thrombus formation. Further developments in gene transfer vectors and gene delivery techniques are required before a full therapeutic potential or gene therapy in cardiovascular diseases can be evaluated. PMID- 8732636 TI - Long-term health risk of primary hyperparathyroidism: the effect of surgery. AB - Primary hyperparathyroidism is a common disease but 90% of sufferers remain undetected. In elderly women its prevalence is about 1%. The disease can cause severe hypercalcaemia and lead to life-threatening symptoms. However, the majority of cases represent milder forms of hyperparathyroidism with minimal symptoms or no symptoms at all, and slight hypercalcaemia which does not progress during follow-up. Surgery is the only effective treatment of primary hyperparathyroidism and most of the symptoms respond favourably to surgery. Nevertheless, the role of surgery has been controversial in the treatment of mini symptomatic patients. Recently it has become evident that primary hyperparathyroidism is associated with increased mortality due mainly to cardiovascular diseases, and it has been shown that surgical treatment is able to diminish or even eliminate this risk of premature death. The extra mortality is connected also to the mild and nonprogressive forms of the disease, and it seems to have a strong correlation with the duration of the disease. This aspect favours active treatment at an early stage of the disease. Conservative treatment can be considered only in old patients with mild and stable clinical course. Primary hyperparathyroidism is an important risk factor causing morbidity and mortality. PMID- 8732637 TI - High-resolution fluorescence in situ hybridization: a new approach in genome mapping. AB - Mapping of the human genome has been a global effort utilizing both genetic and physical mapping techniques. One approach which has greatly facilitated the physical mapping of the human genome is fluorescence in situ hybridization (FISH). Although FISH is by now a well-established technology, new recently developed modifications have enabled an easier use and higher resolution. The high-resolution FISH techniques have given a special impact in positional cloning: searching the functional gene from a chromosomal area where the gene has been genetically localized. New high-resolution FISH techniques include hybridization of probes to free chromatin, DNA fibres or mechanically stretched chromosomes. These targets have widened the resolution of FISH to detect distances from the traditional cytogenetic resolution level down to a resolution of a few kilobases. They also have significantly speeded up high-resolution physical mapping and thus made the search of new disease genes easier. PMID- 8732638 TI - Histamine-2 receptor antagonists as immunomodulators: new therapeutic views? AB - Considerable evidence has emerged to suggest that histamine participates in the regulation of the inflammatory response, immune reaction, coagulation cascade, and cardiovascular function. Furthermore, histamine may play a major role in the growth of normal and malignant tissue as a regulator of proliferation and angiogenesis. Specific histamine receptors have been identified on the surface of bone marrow cells, immune competent cells, endothelial cells, fibroblasts, and also on malignant cells. This has prompted research in regulation by specific histamine receptor agonists and antagonists. Results from such studies are currently accumulating and suggest that the histamine-2 receptor antagonists have potential beneficial effects in the treatment of certain malignant, autoimmune and skin diseases, either alone or in combination with other drugs. The beneficial effect of histamine-2 receptor antagonists as adjuvant single drugs to reduce trauma-, blood transfusion- and sepsis-induced immunosuppression has led to research in combined treatment regimens in major surgery, particularly, of patients operated on for malignant diseases. PMID- 8732639 TI - Bone marrow transplantation in immunodeficiency diseases. AB - Bone marrow transplantation (BMT) has become the treatment of choice in an increasing number of primary immunodeficiency disorders. In many of these otherwise lethal genetic diseases, BMT can completely reverse the immunological abnormality and patients may be permanently cured. In this review, current approaches in the application of this treatment and results obtained in various entities of primary immunodeficiencies are presented. A significant prognostic factor for the outcome is the clinical condition of the patient at the time of BMT, and prompt recognition of these rare disorders remains an important prognostic factor for BMT. PMID- 8732640 TI - Menkes disease: recent advances and new insights into copper metabolism. AB - Copper is a trace element necessary for the normal function of several important enzymes but copper homeostasis is still poorly understood. In recent years remarkable progress has been made in this field following the isolation of the gene defective in Menkes disease. Menkes disease and occipital horn syndrome are X-linked recessive disorders, demonstrating the vital importance of copper, which is also highly toxic in excessive amounts. Its destructive effects are reflected in the autosomal recessive Wilson's disease. Progressive neurodegeneration and connective tissue disturbances are the main manifestations of Menkes disease. Although many patients present a severe clinical course, variable forms can be distinguished, and the occipital horn syndrome has been suggested to be a mild allelic form. The Menkes locus is mapped to Xq13.3 and the gene defective in Menkes disease has been isolated by positional cloning. The gene is predicted to encode an energy-dependent copper-binding protein, the first intracellular copper transporter described in eukaryotes. Isolation of the gene and subsequent characterization of the exon-intron organization now enables the establishment of DNA-based diagnostic methods. Furthermore, identification of the Menkes disease gene led to other important findings, such as isolation of its mouse homologue, confirming the allelic relationship between Menkes disease and occipital horn syndrome, and isolation of the defective genes in Wilson's disease and its rat homologue. PMID- 8732641 TI - MRI in obstetrics: a supplementary method for ultrasonography. AB - The use of magnetic resonance imaging (MRI) as a complementary diagnostic method was evaluated in a group of 36 complicated pregnancies during the last trimester. Most of the examinations were carried out by two specific MRI sequences, the T1 weighted gradient echo FLASH sequence (TR 120, TE 12) and the T2-weighted fast spin echo sequence (TR 2000, TE 90). Information additional to that obtained by ultrasonic diagnosis was obtained by MRI in 16 of the 25 cases of foetal malformation and in 5 of the 11 cases with other obstetric problems. Foetal curarization was not necessary in most of the cases. A good MRI technique, for selected obstetric problems and when used in combination with detailed antepartal ultrasonic examinations, can give useful diagnostic information to an obstetrician. PMID- 8732642 TI - Autologous stem cell transplants in lymphomas. AB - Hodgkin's disease and non-Hodgkin's lymphomas can be treated and, in a large number of cases, cured by first-line chemotherapy or radiotherapy. Unlike many other malignancies, relapse is not uniformly fatal but the treatment is usually markedly myelotoxic with the high doses of chemotherapy used in relapse. Haematopoietic reconstitution with either autologous marrow or peripheral stem cells postchemotherapy has made high-dose chemotherapy relatively safe with mortality rates as low as 2% in some centres. The clinical indications for high dose therapy in lymphoma management for patients with relapsed and bad prognosis disease are reviewed. The advantages of autologous bone marrow and peripheral stem cell transplants are compared and current peripheral stem cell mobilization and harvesting practice is discussed. PMID- 8732643 TI - Autologous stem cell transplantation in breast cancer. AB - High-dose chemotherapy with autologous stem cell support is an investigational treatment for patients with breast cancer who have a high risk of recurrence or who have metastatic disease. In the adjuvant setting, there is a lack of survival data from randomized studies comparing high-dose to conventional-dose therapy. High-dose adjuvant chemotherapy was found to result in as high as 71% 5-year event-free survival in patients with more than nine metastatic axillary lymph nodes in one nonrandomized study, which appears to be superior to those achieved with conventional-dose therapy. In metastatic breast cancer the great majority of patients die of cancer despite high-dose therapy with the current regimens, and it is unclear whether survival of these patients is improved with high-dose therapy as compared to conventional-dose therapy. However, a few patients with breast cancer with distant metastases have enjoyed a disease-free interval lasting for several years after high-dose therapy, which is rarely seen with conventional-dose therapy, and one small randomized trial showed improved survival in the high-dose arm of the study. The more extensive patient selection for high-dose chemotherapy needs to be taken into account when comparing the results achieved with different dose levels, and therefore a high priority should be given for randomized studies where high-dose therapy is compared with conventional-dose therapy. PMID- 8732644 TI - High-dose chemotherapy with autologous stem cell support in advanced ovarian cancer. AB - Since 1981, over 300 patients reported with advanced or refractory ovarian cancer have been treated with high-dose chemotherapy supported by autologous bone marrow or peripheral blood stem cell transplantation. Partial or complete clinical response has been reported in 54-100% of the cases, but the median duration of the response in the majority of patients has been only a few months. It is obvious from the available data that high-dose regimens supported by autologous stem cell transplantation (ASCT) are not capable of inducing long-term survival in patients with heavy tumour burden or chemoresistant ovarian cancer. Recent reports on nearly 100 patients have described results of the use of high-dose chemotherapy as first-line treatment for patients with optimally debulked disease or negative second-look laparotomy. Response rates and survival have been better when compared to historical controls, but the efficacy of this treatment modality in inducing durable remission has not been tested in randomized trials. Most of the ongoing trials presented briefly in this review have been designed to evaluate the potential of high-dose therapy as first-line treatment in preventing the development of resistant tumour clones and recurrence. The role of sequential high-dose chemotherapy with ASCT as a part of primary treatment or as salvage therapy for chemosensitive recurrent disease is also under investigation. PMID- 8732645 TI - Autologous stem cell transplantation in solid tumours of childhood. AB - High-dose chemoradiotherapy (HDCRT) followed by autologous stem cell (ASC) rescue is now widely used in a number of childhood malignancies. The most common to date is neuroblastoma, where it now has an established role as consolidation of initial complete remission in children over the age of 1 year with stage 4 disease. High-dose melphalan alone prolongs progression-free survival, with a small increase in long-term survival. The value of a total body irradiation (TBI) based regimen is currently under randomized evaluation. In soft-tissue sarcoma, such as rhabdomyosarcoma or Ewing's sarcoma of bone or soft tissue, high-dose therapy has been used to consolidate initial complete or partial remission. The benefit has not been demonstrated in randomized studies but chemotherapy-alone regimens based on combined alkylating agents appear to be of potential value. Other tumours have been treated in this way but the place of high-dose therapy remains entirely unclear. It is possible that inherently chemosensitive tumours, such as Wilm's tumour, may be candidates where standard treatment has failed. The replacement of bone marrow reinfusion with cytokine-primed peripheral blood stem cell (PBSC) rescue has reduced the morbidity of these procedures. The issue of the potential risk of reinfusing tumour cells has been addressed in neuroblastoma where purging using immunodepletion or in vitro chemotherapy has been carried out. PMID- 8732646 TI - Purging of autografts: methods and clinical significance. AB - High-dose chemoradiotherapy with autologous haematopoietic progenitor cell support is being used with increasing frequency to treat patients with a variety of malignancies. The most common reason for the ultimate failure in patients treated with high-dose therapy with stem cell support is not lack of engraftment, toxicity of therapy, or infection, but rather relapse of disease. There is solid evidence that reinfusion of autografts containing clonogenic tumour cells can contribute to relapse and influence patient outcome after high-dose treatment. In patients undergoing high-dose treatment, tumour cell contamination can be observed in histologically normal bone marrow or peripheral blood by sensitive micrometastatic detection techniques. Consequently, methods for detection of minimal residual disease in the autografts, together with techniques to remove tumour cells from the harvest are considered to be important in patients receiving high-dose therapy with stem cell support. PMID- 8732647 TI - Incidence and prevention of nonhaematological toxicity of high-dose chemotherapy. AB - The introduction of the haematopoietic growth factors (HGFs), together with the evolution of techniques to harvest haematopoietic stem cells from the peripheral blood, have greatly facilitated the use of high-dose chemotherapy (HDC). While haematological toxicity of HDC is no longer dose-limiting, damage to other tissues has become more pronounced. In fact, nonhaematological toxicity (NHTOX) is now often dose-limiting in HDC regimens. NHTOX associated with HDC regimens depends on the type and dose of the drugs used, the physical condition and the characteristics of the patients treated and the given comedication. We describe the most important toxic effects of commonly used HDC programmes, such as nausea, vomiting, and mucositis, neutropaenic fever and sepsis, various major organ toxicities, catheter-associated problems and long-term complications. In addition, we discuss the possibilities of preventing these side-effects and what action to take if they occur. PMID- 8732648 TI - Fine-needle sampling of salivary gland lesions. I. Cytology and histology correlation of 412 cases of pleomorphic adenoma. AB - Fine-needle sampling (FNS) of 412 pleomorphic adenomas, corresponding to 378 primary tumors and 34 local recurrences, was performed preoperatively in 378 patients. Concordant cytologic diagnoses were established in 376 tumors (91.3%), whereas seven (1.7%) were only classified as benign. Sixteen (3.9%) tumors were considered to be suspicious and two (0.5%) were diagnosed as adenoid cystic carcinomas. Insufficient material for cytologic evaluation was found in 11 (2.6%) tumors. Pleomorphic adenomas were retrospectively classified according to the predominant histological differentiation: chondromyxoid, cellular, myoepithelial, or metaplastic. FNS performances were higher in chondromyxoid than in other types, with 95.2% concordant and 1.4% suspicious/ false-positive, vs. 81.8% concordant, and 11.5% suspicious/false-positive cytodiagnoses, respectively. PMID- 8732649 TI - Use of thin-layer preparations for gynecologic smears with emphasis on the cytomorphology of high-grade intraepithelial lesions and carcinomas. AB - Thin-layer (TL) technology can improve the detection rate for squamous lesions of the uterine cervix. Studies to date have under-represented high grade lesions and malignancies. The present study utilized a patient population at high risk for such lesions in order to analyze the performance of TL procedures in this group, and in addition, to assess the similarities and differences in morphologic appearances of specimens prepared by the two methods. Conventional (CS) and thin layer smears (TLS) were made in parallel from the same specimen. Each slide was examined in a blinded fashion. Diagnoses were compared and morphologic observations made. Two hundred fifty-nine cases were included, of which 32 (12%) were high grade dysplasias (11) or carcinomas (21). Thirty five (14%) were atypical or low grade dysplasias. There was exact correlation between Bethesda classification in 231 cases (89%). Of the 21 carcinomas identified, 19 (91%) were present on each preparation. Two cases of endometrial adenocarcinoma were missed on unsatisfactory or negative TLS. One case of squamous cell carcinoma was called high grade squamous intraepithelial lesion (HGSIL) on TLS while the CS was unsatisfactory. Three cases called atypical glandular cells (AGCUS) on TLS, and negative on CS, showed HGSIL (1) or no lesion (2) on follow-up. Morphologic features of low grade lesions were virtually identical on both preparations. Distinct features were noted on TLS in the high grade lesions. These included smaller appearing nuclear areas, less distinct nuclear chromatin, thicker three dimensional groupings, and more isolated cells. Such findings were most pronounced in the glandular lesions. With training and experience, these features were easily identified in TL preparations, further documenting the utility of this procedure for use in routine practice. PMID- 8732650 TI - Fine-needle aspiration of supraclavicular lymph nodes. AB - Malignancies from many primary sites may metastasize to supraclavicular lymph nodes (SCLN). We reviewed 100 fine-needle aspirations (FNAs) of SCLNs. There were three major types of malignancy detected by this method: adenocarcinoma (n = 40), squamous cell carcinoma (n = 14), and other malignancies (n = 29). Adenocarcinomas and other malignancies from all sites tended to metastasize to the left SCLN. Squamous cell carcinomas from all sites, however, tended to appear on the right side. For 61 patients, a previous diagnosis of malignancy had been made within 1 yr of the clinical appearance of the abnormal SCLN. For 20 patients, the primary diagnosis antedated SCLN metastasis by more than 1 yr, particularly patients with adenocarcinoma of the breast, prostate, or thyroid papillary carcinoma. PMID- 8732651 TI - Contribution of immunocytochemistry in routine diagnostic cytology. AB - The authors report the experience of applying immunocytochemistry in routine cytological examination and its contribution for diagnosis during a period of 3 yr in Veterans General Hospital-Taipei, Taiwan. From August 1991 to July 1994, the cytology laboratory received 5,762 non-gynecologic specimens with urine excluded. Immunocytochemistry was performed selectively in problematic cases. A total of 215 stainings including 124 epithelial markers, 50 lymphoma/leukemia markers, 22 neuroglial and choroid plexus markers, seven mesenchymal markers, six melanoma markers, and six others was performed on 145 specimens consisting of 89 effusions, 28 fine-needle aspirations, 11 cerebrospinal fluids, and 17 other specimens. Effusions were by far the most frequent specimens for immunocytochemistry and the epithelial markers were the most frequently used antibodies. The immunocytochemical results were essential in 41 specimens (28%), confirmatory in 37 (26%), and non-contributory in 67 (46%). Essential and confirmatory results occurred in 49% of effusions (44/89), 71% of fine-needle aspirations (20/28), 55% of cerebrospinal fluids (6/11), and 47% of other specimens (8/17). It is concluded that immunocytochemistry is proved to be a good aid for the final diagnosis of daily cytologic practices in which the fine-needle aspiration specimens are benefitted best. PMID- 8732652 TI - Metaplastic breast carcinoma: a cytohistologic and clinical study of 10 cases. AB - Metaplastic breast carcinomas (MBC) account for < 5% of breast malignancies and have an uncertain prognostic significance. The tumors tend to grow rapidly, and most are negative for hormone receptors. Adenosquamous carcinomas and various types of carcinosarcomas may be classified as MBC. We evaluated the cytologic, histologic, and clinical parameters of 10 MBC to determine important diagnostic features of these tumors. A cytologic diagnosis of MBC, based on the identification of two distinct malignant components, was made preoperatively in five of 10 (50%) cases, and retrospectively in two additional cases; two specimens were inadequately cellular. Poorly-differentiated adenocarcinoma was the most frequently encountered component of MBC. It is recommended that malignant breast aspirates be carefully scrutinized for multiple neoplastic components. Our series of MBC differs from previous reports in that two cases presented as inflammatory breast cancer, one case was pregnancy-associated, and there was a higher incidence of estrogen and progesterone receptor positivity. No cancer-related deaths occurred during a mean follow-up period of over 6 yr. PMID- 8732653 TI - Fine-needle sampling of a case of carcinoma of the breast with neuroendocrine differentiation. AB - Fine-needle sampling was performed in a woman with a left breast lump. The cytologic diagnosis was consistent with a poorly-differentiated carcinoma. Cytologic features included medium-to-large, round, and spindle-shaped cells with scanty cytoplasm, nuclear molding, and rosette-like structures. Histology revealed an endocrine pattern. Immunohistochemical staining was positive for epithelial and neuroendocrine markers, and electron microscopy showed many small membrane-bound electron-dense granules, confirming the diagnosis of breast carcinoma with neuroendocrine differentiation. DNA flow cytometry and cytogenetic analyses revealed a near-tetraploid tumor. PMID- 8732654 TI - Fine-needle aspiration cytology of spindle-cell argyrophilic mucin-producing carcinoma of the breast. AB - An unusual breast carcinoma with prominent spindle cells, intracytoplasmic mucin, and argyrophilia in an 82-yr-old woman was sampled preoperatively by fine-needle aspiration (FNA). The smears contained highly cellular sheets and fragments of uniform spindle cells with abundant vacuolated cytoplasm. The cells were mucicarmine positive and contained many cytoplasmic argyrophilic granules. The findings were confirmed by histologic examination of the excised tumor, which was also studied with a panel of immunohistochemical stains. A review of the literature indicated that spindle-cell argyrophilic mucin-producing carcinoma (SCAMPC) of the breast is a very rare neoplasm presenting primarily in elderly patients. The FNA cytologic features seem to be unique and should allow distinction from other primary breast lesions. PMID- 8732655 TI - Concomitant malignant mesothelioma of the pleura, peritoneum, and tunica vaginalis testis. AB - We describe the cytohistological, immunohistochemical and ultrastructural findings in a 55-yr-old-man with history of asbestos exposure and diffuse malignant mesothelioma (DMM) of the pleura, peritoneum, and tunica vaginalis presenting with chest pain and scrotal swelling. Pleural fine-needle aspiration (FNA) revealed mesenchymal elements and spindle-shaped epithelial-like cells, while biopsy showed pure sarcomatous tumor invading lung parenchymal. In both samples tumor cells coexpressed cytokeratin and vimentin. Peritoneal and hydrocele effusions contained aggregates of malignant mesothelial cells. Electron microscopy showed intermediate filaments, rare desmosomes and sparse microvilli. Morphological findings were consistent with a DMM, with a biphasic pattern in the pleura and an epithelial one in the peritoneum and tunica vaginalis. Although the possibility of a multicentric origin cannot be ruled out, clinical chronologic sequence suggests that the pleura was the primary involved site, followed by spread to peritoneum and tunica vaginalis. PMID- 8732656 TI - Fine-needle aspiration cytology of ameloblastic carcinoma of the maxilla: a rare tumor. AB - This report describes the cytologic, histologic, and clinical features of an ameloblastic carcinoma of the maxilla occurring in an 83-yr-old white male. A fine-needle aspiration revealed malignant cells with a predominantly small cell type morphology. There were also a few cells of a second type which were more polygonal to spindled with oval to elongate nuclei. A focal amorphous blue-grey matrix was noted in association with these cells. Although the biopsy showed prominent areas of squamous metaplasia, no squamous cells were seen in the cytologic sample. PMID- 8732657 TI - Cytologic findings in a case of T-cell rich B-cell lymphoma: potential diagnostic pitfall in FNA of lymph nodes. AB - Fine-needle aspiration biopsy (FNAB) is a reliable diagnostic technique for most palpable masses. This technique is utilized routinely to diagnose metastatic carcinoma and melanomas in lymph nodes. However, the role of FNAB in the investigation of lymphoproliferative lesions is still controversial. Recent publications have supported the use of FNAB cytology, in conjunction with immunophenotyping, as an accurate, reliable diagnostic modality for the classification of most lymphomas (Sneige et al., Acta Cytol 1990; 34:311-322; Skoog and Tani, Diagn Oncol 1991; 1:12-18; Robins et al., Am J Clin Pathol 1994; 101:569-576; Katz, Clin Lab Med 1991; 11:469-499). We present a case of a T-cell rich, large B-cell lymphoma. Material obtained by FNAB mimicked a reactive process by both cytomorphological and immunophenotypical analysis. This case demonstrates a potential pitfall in the use of FNAB to evaluate lymphoproliferative disorders even when used in conjunction with immunophenotypic studies. The case also emphasizes the need for detailed clinical and prior pathologic information when a cytologic sample is being evaluated for a lymphoproliferative disorder. To our knowledge, the cytomorphologic findings of this particular type of lymphoma have not been previously described as seen on an FNAB. PMID- 8732658 TI - Cytomorphology of a mixed pigmented/nonpigmented pleomorphic adenoma of the ciliary body: a diagnostic pitfall in the evaluation of intraocular tumors. AB - Intraoperative cytology is an alternative and sometimes preferred method to frozen section in the rapid evaluation of surgical biopsies. This report describes the cytological, histological, and immunohistochemical features of a case of mixed pigmented/nonpigmented pleomorphic adenoma of the ciliary body with clinical and morphological features reminiscent of malignant melanoma. The characteristic cyto-histomorphology of this unusual tumor and its distinguishing features from malignant melanoma are discussed. PMID- 8732659 TI - Fine-needle aspiration cytology of bronchocentric granulomatosis: a potential diagnostic pitfall. AB - Bronchocentric granulomatosis (BCTG) is a rare disease associated with bronchial asthma and bronchopulmonary aspergillosis. Idiopathic forms are rarely encountered. We report on a case of BCTG showing clinical, radiological, and cytological evidence suggestive of adenocarcinoma of the lung. The patient is a 69-yr-old female, lifetime nonsmoker with multiple sclerosis who was admitted with a history of ascending cholangitis. Admission chest X-ray documented a 1.5 cm nodule in the left upper lobe of the lung. This was confirmed by CT scan. The lesion was slowly growing. Bronchoscopic examination was normal. Bronchial brushings were inconclusive. A transthoracic fine-needle aspiration showed sheets of highly atypical epithelium with occasional small dyshesive clusters. There was an inflammatory background that was believed to represent tumor diathesis. The cytological interpretation was "suspicious for adenocarcinoma." The patient underwent left upper lobectomy. The lung showed multiple peribronchial granulomas with intense peribronchial lymphoid infiltrate extending into the bronchial mucosa, causing cytological atypia and focal ulceration. Special stains for microorganisms were negative. The patient recovered from surgery and shows no signs of infection. We conclude that BCTG and related lesions can give cytological features that are suggestive of malignancy. Cytological material obtained from these lesions should be interpreted with caution. PMID- 8732660 TI - Fine-needle aspiration as the initial diagnostic modality in malignant lung disease. AB - Cytologic detection of lung cancer is accepted, accurate, and time-honored. Typically, cytologic workup of a radiologic abnormality proceeds sequentially from sputum to bronchoalveolar cytology, and, if necessary, to fine-needle aspiration biopsy (FNA). Initial use of FNA in lung cancer diagnosis is controversial, but increasingly popular. We therefore decided to objectively assess current practice in cytologic lung cancer diagnosis at our institution. All pulmonary cytologic diagnoses for 1993 and the first half of 1994 were retrieved. Positive diagnoses were then used to access all patient data. Patients were stratified according to the specimen from which the first positive diagnosis was obtained. Of 542 pulmonary cytology specimens, 15% were sputa, 65% were bronchoalveolar, and 20% were FNAs. One hundred sixty-one of 172 malignant diagnoses were first diagnoses. Three percent of first malignant diagnoses were made from sputa, 47% were from lavages, and 50% were from FNAs. Although FNAs comprised just 20% of all pulmonary cytologies, 50% of all new malignant cytologic diagnoses were made by FNA. Initial use of FNA is successful, has a high diagnostic yield and low complication rate, and offers the most direct approach to diagnosis. PMID- 8732661 TI - Detection of Trichomonas vaginalis in endocervical and ectocervical smears. AB - The objective of this study was to determine the relative sensitivity of detecting Trichomonas vaginalis in endocervical as compared to ectocervical Papanicolaou-stained smears. The average number of organisms in 10 (200x) high power fields (hpf) per smear was used for comparing the presence of trichomonads in the two smear types. In every pair of ectocervical-endocervical smears, there were always more trichomonads in the ectocervical smear. Fifty percent of cases showed at least 25 more trichomonads per 200x hpf in the ectocervical smear than in the endocervical smear. In 20 of 50 cases (40%), trichomonads were present only in the ectocervical smear. The organisms were found only in the endocervical smears of women with a heavier ectocervical trichomonad burden. In conclusion, trichomonads are detected in the Papanicolaou-stained ectocervical smear more often than in the endocervical smear; therefore, examination of an adequate ectocervical smear has greater utility than the endocervical smear for the diagnosis of trichomoniasis. PMID- 8732662 TI - Cell block techniques and immunocytochemistry. PMID- 8732663 TI - Quality assurance cytology error tracking system for negative GYN case rescreen. PMID- 8732664 TI - Establishing standards of care. PMID- 8732665 TI - Phosphatidylinositol 3-kinase recruitment by p185erbB-2 and erbB-3 is potently induced by neu differentiation factor/heregulin during mitogenesis and is constitutively elevated in growth factor-independent breast carcinoma cells with c-erbB-2 gene amplification. AB - Amplification and overexpression of the c-erbB-2 gene in 21MT-2 and 21MT-1 human breast carcinoma cells results in progressively elevated levels of constitutively tyrosine-phosphorylated p185erbB-2 and is associated with progressive insulin like growth factor (IGF) and combined IGF/epidermal growth factor (EGF) independence in culture. In addition, the neu differentiation factor/heregulins (HRGs), a family of ligands that activate p185erbB-2 through direct binding to erbB-3 or erbB-4, are potent mitogens for various nonneoplastic mammary epithelial cells and carcinoma cell lines in the absence of both IGF and EGF in culture. We have investigated the ability of ligand induction with HRGs or the constitutive activation of p185erbB-2 in the 21MT breast carcinoma cells to induced the recruitment of phosphatidylinositol 3-kinase (PI3K) by p185erbB-2 and erbB-3. HRG was found to potently induce the recruitment of the M(r) 85,000 regulatory subunit of PI3K by phosphotyrosine proteins in both nonneoplastic H16N 2 mammary epithelial cells (which express normal c-erbB-2 levels) and in the 21MT 2 and 21MT-1 cell lines, which were all isolated from a single patient with intraductal and invasive ductal carcinoma of the breast and express c-erbB-3 but not c-erbB-4 in culture. The activation of PI3K in these cells was also associated with high-level mitogenic responsiveness to HRG, as well as the IGF/EGF-independent proliferation of the 21MT cell lines in culture. The recruitment of PI3K by phosphotyrosine protein during ligand-induced activation, or that seen constitutively in the 21MT tumor cells, did not involve detectable tyrosine phosphorylation of p85. The HRG-induced recruitment of p85 and the constitutive recruitment of p85 in the 21MT cell lines involved direct association with both p185erbB-2 and erbB-3, although greater levels were recruited directly by erbB-3. Wortmannin, a potent inhibitor of PI3K enzymatic activity, also blocked the autonomous proliferation of the 21MT cells, and this effect was reversible in long-term cultures. These data indicate that PI3K may be an especially important mediator of HRG-induced proliferation in mammary epithelial cells and is involved in the autonomous proliferation of growth factor independent breast carcinoma cells with c-erbB-2 gene amplification. PMID- 8732666 TI - Nonconstitutive expression of the gastrin-releasing peptide autocrine growth system in human small cell lung carcinoma NCI-H345 cells. AB - Constitutive, unregulated autocrine growth is thought to be an important mechanism whereby cancer cells gain a proliferative advantage over nonmalignant cells. The question addressed here was whether the autocrine growth system for gastrin-releasing peptide (GRP) in human small cell lung carcinoma cells is, in fact, always expressed in a constitutive, unregulated fashion. Lag, rapid, and plateau growth states were defined for small cell lung carcinoma NCI-H345 cells based on periods during which they expressed different growth rates after plating as single cell suspensions. Immunoreactive GRP in the conditioned medium and in NCI-H345 cells harvested during each of these growth states, as well as cell DNA content, GRP mRNA expression, specific 125I-GRP uptake, specific 125I-GRP binding to solubilized membranes, and GRP and neuromedin B receptor mRNA expression by reverse transcription-PCR were analyzed. Maximal levels of GRP expression were observed during the lag growth state, with the highest concentration of immunoreactive GRP in the conditioned medium during the rapid growth state. Specific 125I-GRP uptake and binding were also highest during the lag growth state; however, GRP receptor mRNA did not significantly change. In contrast to prevailing concepts, these studies support the conclusion that the expression of the GRP autocrine growth system in NCI-H345 cells is indeed regulated. Furthermore, the components are maximally expressed before rapid growth begins, suggesting that other mechanisms are activated to support the actual proliferation. PMID- 8732667 TI - Endogenous fibroblast growth factor-1 or fibroblast growth factor-2 modulate prostate cancer cell proliferation. AB - We constructed expression vectors containing either rat fibroblast growth factor (FGF)-1 for FGF-2 cDNA cloned in either the sense orientation or antisense orientation relative to the metallothionein promoter of plasmid pMTneo.1. Stable AXC/SSh rat prostate cancer cell transfectants expressing either chimeric FGF-1 sense, chimeric FGF-1-antisense, or chimeric FGF-2-antisense transcripts were obtained. Stable transfectants expressing chimeric FGF-2-sense transcripts were not obtained. Control, sense, and antisense transfectants expressed endogenous FGF-1 and endogenous FGF-2 transcripts, implying that transfection did not eliminate endogenous FGF transcripts. Control transfectants and sense transfectants contained FGF-1 isoforms having a mass of 16.4 or 17.3 kDa and FGF 2 isoforms having a mass of 17, 19.5, or 21.5 kDa. Significantly, adult AXC/SSh rat prostate contained only the 17.3 kDa FGF-1 isoform and the 17 kDa FGF-2 isoform, indicating that neoplastic transformation was associated with elaboration of novel, prostate epithelial cell-derived FGF-2 isoforms. FGF-1 antisense RNA expression eliminated transfectant FGF-1 isoforms without affecting FGF-2 isoform content. Similarly, FGF-2-antisense RNA expression eliminated the transfectant 21.5 kDa FGF-2 isoform, diminished the 19.5 kDa FGF-2 isoform content, and reduced the 17 kDa FGF-2 isoform content to barely detectable levels without affecting the FGF-1 isoform content. This established that FGF-antisense RNAs specifically inhibited translation of cognate, endogenous FGF transcripts. Doubling times of control transfectants and sense transfectants were indistinguishable and were not affected by including FGF-1 or FGF-2 in the culture medium. Doubling times of FGF-1-antisense or FGF-2-antisense transfectants were 1.3- to 1.4-fold greater than those of control transfectants or sense transfectants, and either exogenous FGF-1 or exogenous FGF-2 decreased antisense transfectant doubling times to values indistinguishable from those of control transfectants or sense transfectants. This established that with regard to prostate cancer cell proliferation: (a) endogenous FGF-1 cannot substitute for endogenous FGF-2 eliminated by FGF-2-antisense RNA expression; and (b) endogenous FGF-2 cannot substitute for endogenous FGF-1 eliminated by FGF-1-antisense RNA expression. In contrast, either exogenous FGF-1 or exogenous FGF-2 decreased antisense transfectant doubling time. The results of these studies establish that endogenous FGF-1 and endogenous FGF-2 modulate prostate cancer cell proliferation and imply that FGF-1 and FGF-2 of endogenous and exogenous origin conjointly control aspects of prostate cancer cell homeostasis. Our findings suggest complex interaction between components of prostate cancer cell regulatory processes and endogenously produced and exogenously accessible FGF-1 and FGF-2. PMID- 8732668 TI - Protein kinase C beta 1 and protein kinase C beta 2 activate p57 mitogen activated protein kinase and block differentiation in colon carcinoma cells. AB - When HD3 colon carcinoma cells differentiate to fluid-transporting, enterocytic like cells, they down-regulate their protein kinase C (PKC) beta levels 5-10-fold and lose two responses to basic fibroblast growth factor (FGF): proliferation and the ability to activate p57 mitogen-activated protein (MAP) kinase. HD3 cells were transfected with expression plasmids for the splice variants PKC-beta 1 and PKC-beta 2 and the empty vector for a control. Each of two PKC-beta 1 and each of two PKC-beta 2 transfectant clones exhibited elevated levels of Ca(2+)-and phosphatidylserine-dependent PKC activity. Both PKC-beta 1 transfectant clones had elevated levels of PKC-beta 1 protein compared with the PKC-beta 2 transfectants or controls, whereas both PKC-beta 2 transfectant clones had elevated levels of PKC-beta 2 protein compared with PKC-beta 1 transfectants. Control transfectants had no detectable PKC-beta 2 protein. Similar levels of PKC alpha were found in all lines. Each PKC-beta transfectant was less differentiated than the parental line and had regained proliferative response to basic FGF. Increased growth rates in athymic mice were seen for PKC-beta 2 and PKC-beta 1 transfectant cells. Immunocytochemistry of the sectioned tumors showed enhanced protein levels of PKC-beta 2 and PKC-beta 1, correlating increased levels of these isonzymes with increased growth. Increased myelin-basic protein (MBP) kinase activities of M(r) 44,000, 57,000, 63,000, 110,000, and 130,000 by in-gel kinase assay characterized each PKC-beta transfectant. Both Western blotting and immunoprecipitation studies from 35S-prelabeled cells with a pan-erk antibody showed no increase in protein abundance of MAP kinases of M(r) 44,000, 57,000, and 63,000, suggesting that elevated PKC-beta levels led to activation of the smaller three MAP and MBP kinases. Activation of p57 MAP kinase in each PKC-beta transfectant was demonstrated by immunoprecipitation with an antiphosphotyrosine monoclonal antibody and then by assay of the immunoprecipitates by in-gel kinase assay on MBP. p57 MAP kinase was distinguished from the M(r) 54,000 stress activated protein kinases, which migrated more rapidly on SDS gels and could be detected by in-gel kinase assay on MBP only after cellular stress. Thus, expression of elevated levels of PKC-beta 1 and PKC-beta 2 in differentiated HD3 colon carcinoma cells blocked their differentiation, enabled them to proliferate in response to basic FGF like undifferentiated cells, increased their growth rate in athymic mice, and activated several MBP kinases, among them, p57 MAP kinase. PMID- 8732669 TI - Retinoic acid-induced transition from protein kinase C beta to protein kinase C alpha in differentiated F9 cells: correlation with altered regulation of proto oncogene expression by phorbol esters. AB - Retinoic acid (RA) induced differentiation of F9 embryonal carcinoma cells is accompanied by changes in cellular responsiveness to extracellular signals. These changes include an increase in the AP1 transcription factor that is associated with the expression of differentiation markers (e.g., cytokeratin 18 and plasminogen activator). Since AP1 activity is a target for protein kinase C (PKC) regulated changes in gene expression, we have examined the effects of RA on the expression and function of the PKC isozymes. F9 stem cells express PKC beta, delta, epsilon, and zeta. RA-induced differentiation to primitive endoderm led to a transition from PKC beta to PKC alpha expression. Additional treatment with dibutyryl cyclic AMP (dbcAMP), required for terminal differentiation into parietal endoderm, further increased PKC alpha expression and total PKC activity. RA and dbcAMP had negligible effects on the expression of PKC delta, epsilon, and zeta. The PKC beta to PKC alpha transition was specific for parietal endoderm; aggregation of RA-treated F9 cells induced visceral endoderm differentiation with elevated expression of PKC beta. The PKC activation with phorbol esters induced the expression of c-fos, c-jun, and junB proto-oncogenes in F9 stem cells. In the presence of either RA or RA and dbcAMP, phorbol ester treatment enhanced the expression of type IV collagen, a parietal endoderm marker. It also increased the expression of c-jun gene but not c-fos. The specific involvement of PKC beta in c fos induction and PKC alpha in type IV collagen induction was confirmed in each PKC isozyme-transfected F9 cells. Together, our data demonstrate that the RA induced (and dbcAMP-induced) changes in conventional PKC expression alters gene expression during parietal endoderm formation. PMID- 8732670 TI - Effect of 1-beta-D-arabinofuranosylcytosine on apoptosis and differentiation in human monocytic leukemia cells (U937) expressing a c-Jun dominant-negative mutant protein (TAM67). AB - The proto-oncogene c-jun encodes a component of the AP-1 transcription-activating complex and has been implicated in the regulation of diverse cellular processes, including cell proliferation, differentiation, transformation, and most recently, apoptosis. We have used a U937 monocytic leukemia cell line stably expressing a c jun dominant-negative, transactivation-domain deletion mutant (TAM67) to assess the role of c-jun in apoptotic events induced by exposure to the antimetabolite 1 beta-D-arabinofuranosylcytosine (ara-C). Mutant cells produce a truncated M(r) 29,000 protein that interferes with the function of normal c-Jun (and c-Fos) proteins through a quenching mechanism. Parental U937, cells expressing TAM67, and cells carrying only the empty vector (pMM) were exposed to ara-C for 6 h, and apoptosis was monitored by cell morphology as well as qualitative and quantitative assays of DNA damage. No differences in apoptosis could be detected between the three cell lines at any of the ara-C concentrations evaluated. In addition, ara-C concentrations > or = 1.0 x 10(-6) M were equally inhibitory to the clonogenic growth of U937 and TAM67-expressing cells. In contrast, lower concentrations of ara-C (i.e., < 5.0 x 10(-7) M) were significantly less inhibitory to mutant U937 cell colony formation than to their parental counterparts. The reduced sensitivity of TAM67-expressing cells to low concentrations of ara-C could not be attributed to biochemical or cytokinetic factors, since the two cell lines were indistinguishable with respect to 1-beta-D arabinofuranosylcytosine 5'-triphosphate (ara-CTP) formation, ara-CTP:dCTP ratios, and S-phase fraction. However, a significantly lower percentage of TAM67 expressing cells exposed to submicromolar concentrations of ara-C exhibited features associated with a differentiated monocytoid phenotype (i.e., increased plastic adherence and CD11b expression) compared to their parental counterparts. Lower concentrations of ara-C were also significantly less effective in decreasing the percentage of S-phase cells and in down-regulating c-myc mRNA levels in the mutant line, events associated with induction of leukemic cell differentiation. Finally, ara-C-induced up-regulation of c-jun message and protein was markedly attenuated in TAM67-expressing cells, findings consistent with a c-jun dominant-negative model. Collectively, these findings suggest that dysregulation of c-jun in U937 cells antagonizes low-dose ara-C-mediated cellular maturation but does not prevent higher concentration of this agent from triggering apoptosis. They also raise the possibility that separate aspects of the antiproliferative actions of ara-C may be differentially regulated by c-jun. PMID- 8732671 TI - Selective changes in laminin adhesion and alpha 6 beta 4 integrin regulation are associated with the initial steps in keratinocyte maturation. AB - In skin, the distribution of integrins is compartmentalized. Whereas the alpha 6 beta 4 integrin complex is polarized to the basal portion of proliferating cells in the basal layer juxtaposed to the basement membrane, alpha 3 beta 1 integrin receptors are localized on the cell surface surrounding basal and suprabasal cells, suggesting beta 1 integrins mediate both cell-matrix and cell-cell interactions. As initiation of maturation in skin is associated with the detachment of cells from the basement membrane, the early loss of alpha 6 beta 4, but not alpha 3 beta 1, integrin expression could be a determining factor in the transition from the proliferating to a differentiating keratinocyte. We have studied the regulation of adhesion potential and integrin expression during differentiation of mouse basal keratinocytes culture in 0.05 mM Ca2+ medium and induced to differentiate in 0.12 mM Ca2+ medium. Within 12-24 h after elevation of Ca2+, a selective loss of the alpha 6 beta 4 integrin complex is associated with the induction of the spinous cell marker keratin 1. This early differentiation phenotype coincides with loss of cell attachment mediated by alpha 6 beta 4 to laminins 1 and 5 but not a fibronectin or collagen IV. Selective loss of attachment to laminin is also detected in spinous cells isolated from newborn epidermis in vivo. The loss of alpha 6 and beta 4 protein expression is a consequence of transcriptional and posttranscriptional events, including reduction in mRNA transcripts, reduced synthesis of the alpha 6 protein, and enhanced processing of the alpha 6 and beta 4 chains as determined by Western blots and pulse-chase experiments in metabolically labeled keratinocytes. Selective processing of the beta 4 intracellular domain is detected before loss of beta 4 from the cell surface in basal keratinocytes, and this process is accelerated during differentiation. Whereas early keratinocyte maturation is linked to the selective loss of the alpha 6 beta 4 complex, loss of both beta 1 and beta 4 integrin mRNA and protein occurs as cells proceed to later stages in the differentiation program as induced by 0.5 mM Ca2+ or suspension culture. These conditions are characterized by accelerated expression of transglutaminase; reduced keratin 1 protein; loss of adhesion to fibronectin, laminin 1, laminin 5, and collagen IV; and rapid cell death. Contributing to the down-regulation of beta 1 integrins during terminal differentiation is a selective sensitivity of alpha 3 beta 1 but not alpha 6 beta 4 to down-regulation by transforming growth factors beta 1 and beta 2, factors that are also expressed differentially in normal skin. This study indicates that down-regulation of the alpha 6 beta 4 but not beta 1 integrins occurs during the initial steps of keratinocyte differentiation and is associated with detachment from the laminin matrix. Such changes could contribute an important signal to initiate the process of terminal keratinocyte differentiation. PMID- 8732672 TI - Down-regulation of fibronectin gene expression by the p53 tumor suppressor protein. AB - The tumor suppressor p53 protein down-regulates in vitro the expression of several cellular and viral promoters. However, it is not clear whether this down regulation reflects equivalent modulation of the activity of these promoters in vivo. Here, we propose a suitable system to assess the effect of p53 on gene expression in vivo: the pair of p53 antisense-transfected and parental HeLa cells. The low amount of free wild-type p53 in HeLa cells seems still sufficient for the repression of several promoters that might be derepressed in p53 antisense-transfected HeLa cells. We have used this system for the demonstration both in vivo and in vitro of the repression of the fibronectin (FN) gene promoter by wild-type p53. The protein and mRNA amounts for FN were increased in the p53 antisense-transfected HeLa clones. This was accompanied by the restoration of the FN network in these cells. FN promoter constructs fused to the chloramphenicol acetyltransferase reporter gene were specifically repressed by wild-type p53 in different cell lines. Integrin alpha 5 beta 1 clustering was changed in the sites of focal contacts, most probably representing its relocalization as a consequence of the increased amounts of fibronectin. PMID- 8732673 TI - Focal altered compartmentation of repetitive B2 (Alu-like) sequences in rat liver following hepatocarcinogen exposure. AB - Rats were treated with low doses of the hepatocarcinogens dimethylnitrosamine or thioacetamide, and livers were examined 48 h later. These treatments are known to produce altered RNA compartmentation, wherein a class of repetitive RNA sequences normally restricted to the nucleus appears in the cytoplasm. Reverse transcription-PCR amplifications demonstrated that the sequences showing altered compartmentation consisted largely of a subfamily of the rodent B2 sequence family, the counterpart of human Alu sequences involved in retrotransposition. Northern blot analyses showed that these B2 sequences were found in cytoplasmic RNA as 170- to 360-nucleotide "sense" transcripts, and competition hybridization experiments established that B2 sequences represented most (if not all) of the sequences showing altered compartmentation. The major increase in B2 transcriptions in cytoplasmic RNA was not associated with any change in B2 transcription by RNA polymerase III. In situ hybridizations showed that the altered compartmentation of B2 sequences occurred in well-delineated foci within the rat liver; these foci consisted of a central region containing a prominent infiltrate of macrophages admixed with small hepatocytes and a peripheral region of histologically normal hepatocytes that showed evidence of oxidative damage. Altered compartmentation of B2 sequences may represent an important focal initiatory change in a subset of hepatocytes, whereas subsequent retrotranspositional events (associated with Alu-like sequences) could predispose initiated cell foci to alterations in promotion/progression phases. PMID- 8732674 TI - Cellular growth state differentially regulates enterocyte gene expression in butyrate-treated HT-29 cells. AB - Enterocyte differentiation occurs at the crypt-villus junction through the transcriptional activation of cell-specific genes, including the brush-border enzyme intestinal alkaline phosphatase (IAP) and the microvillar structural protein, villin. Based upon previous in vivo studies demonstrating that IAP and villin are differentially affected by alterations in epithelial growth state, we examined the regulation of these two genes in an in vitro model of enterocyte differentiation. HT-29 cells were maintained in DMEM + 10% FCS and treated with sodium butyrate to induce enterocyte differentiation. Cell count and [3H]thymidine measurements confirm that sodium butyrate causes a marked decrease in cellular growth state, consistent with the differentiation process. Northern blot analyses revealed time- and dose-dependent increases (P < 0.001) in steady state IAP and villin mRNA levels. The increases in both IAP and villin expression were dependent upon the presence of one or more newly synthesized proteins. Post confluence and serum starvation were used to cause cell cycle withdrawal prior to the differentiation process. As predicted from our previous in vivo data, villin expression was unaffected by changes in cellular growth state, whereas IAP expression was dramatically diminished under hypoproliferative conditions. We conclude that early withdrawal from the cell cycle alters the course of enterocyte differentiation, specifically decreasing IAP expression. PMID- 8732675 TI - Radical differences in functions of closely related members of the human carcinoembryonic antigen gene family. AB - The immunoglobulin superfamily represents an ancient, highly diversified group of cell surface and extracellular molecules responsible for a wide range of molecular and cellular recognition functions. The human carcinoembryonic antigen (CEA) subfamily of the immunoglobulin superfamily presents evidence of continuing diversification of the immunoglobulin family, in that some of its members, including CEA itself and nonspecific cross-reacting antigen (NCA), are expressed only in primates and not in rodents. These "new" members are glycophosphatidylinositol linked to the external cell membrane and are up regulated in cancer, unlike members present in both rodents and primates, i.e., biliary glycoprotein (BGP), which are transmembrane linked and down-regulated in cancer. CEA, NCA, and BGP have all been shown to function in vitro as intercellular adhesion molecules. We show here that the properties of adhesion are radically different, in that BGP-mediated adhesion is reversibly Ca2+ and Mg2+ dependent, temperature dependent, and ATP inhibitable, whereas CEA- and NCA mediated adhesion is the opposite in all aspects. Also, the novel double reciprocal, antiparallel binding observed for CEA-CEA interactions is not seen for BGP. Finally, the myogenic differentiation block demonstrated for the ectopic expression of CEA in myoblasts was also observed for NCA but not for BGP, which is consistent with the changes in expression seen in cancer. The appearance of new CEA family members with such different properties is discussed in the context of evolution and cancer. PMID- 8732676 TI - Developmentally regulated expression of metastasis-associated antigens in the rat. AB - The specificity of monoclonal antibodies (mABs) obtained after immunization with a metastasizing rat tumor line was evaluated by screening expression in a variety of nonmetastasizing and metastasizing rat tumor lines. mABs, which by immunohistology and Western blotting recognized metastasizing lines, were used to define the physiological expression of the corresponding antigens during ontogeny as well as in adult rats. From a panel of 12 mABs, 2 recognized structures on metastasizing and nonmetastasizing tumor lines, while 10 stained exclusively metastasizing lines. Five of the latter bound to tumor lines metastasizing either hematogenously or via the lymphatic system. All five recognized an epitope on CD44 variant exon v6. The five remaining mABs, recognizing four independent antigenic entities, only stained tumor lines metastasizing via the lymphatics. Surprisingly, these antigens were also detected in normal tissues: three on epithelial cells either widespread or of the upper gastrointestinal tract or the urogenital system, the fourth preferentially on epithelial cells, but also on nerves and hematopoietic precursor cells, and the fifth on many tissues and cells with a predominance of mesenchyme-derived structures. Notably, during ontogeny, expression on these five antigens was induced in different compartments of the developing fetal and/or maternal part of the placenta. The five newly described metastasis-associated antigens share with CD44v the absence of expression on nonmetastasizing tumor lines as well as expression on distinct, nontransformed cells and induction of expression during ontogeny. Thus, tumor progression may rather be initiated by inappropriate expression or up-regulation of genes, which do not display transforming features, than by de novo appearance of "metastasis genes." Accordingly, metastasizing tumor lines may be a valuable tool to identify developmentally regulated gene products. PMID- 8732677 TI - Altered epidermal cell growth control in vivo by inducible expression of transforming growth factor beta 1 in the skin of transgenic mice. AB - An inducible bovine KIV* keratin gene promoter was used to target expression of latent or activated transforming growth factor beta 1 (TGF beta 1) to keratinocytes in transgenic mice. This short (2.2-kb) keratin 6 (K6) promoter element was generally silent in untreated animals but was induced in keratinocytes when placed in culture or, in vivo, in response to hyperplasia that follows topical application of the tumor promoter, 12-O-tetradecanoylphorbol-13 acetate. All of the K6-TGF beta 1 transgenic lines studied showed attenuation of the basal keratinocyte proliferative response to 12-O-tetradecanoylphorbol-13 acetate as a consequence of inducible TGF beta 1 gene expression. One of the six lines studied showed constitutive transgene expression at low levels in the skin, and this line had a 2- to 3-fold increase in epidermal DNA labeling index over control mice. Although in vitro TGF beta 1 is known to be a potent negative regulator of epithelial cell proliferation, in vivo TGF beta 1 has complex biological activities and can act as either a positive or negative regulator of keratinocyte proliferation. PMID- 8732678 TI - A negative tyrosine aminotransferase gene element that blocks glucocorticoid modulatory element-regulated modulation of glucocorticoid-induced gene expression. AB - Tyrosine aminotransferase (TAT) is the prototypic steroid-inducible gene. Recently, we have found that the modulation of TAT induction properties is reproduced by a novel cis-acting TAT gene element, the glucocorticoid modulatory element (GME). This GME lies about 1 kb upstream of the glucocorticoid response elements (GREs) of the TAT gene and binds a heterooligomer of two recently defined proteins. We now report the existence of an additional TAT gene element between the GME and the GREs that blocks the action of the GME and thus prevents the left shift in the glucocorticoid dose-response curve caused by the GME. This negative element has the properties of a silencer because its activity is relatively position- and orientation-independent. The interaction appears to be stoichiometric in that the effects of a single negative element can be overcome by a second GME. This negative element also has an intrinsic inhibitory activity in the absence of the GME. The majority of the negative element activity could be elicited by a 56-bp sequence between -3105 and -3050 bp of the TAT gene. Multiple, clustered mutations of this sequence reduced, but did not eliminate, the negative activity. Further efforts to restrict the negative element were unsuccessful, suggesting that multiple sequences are required for full activity. High affinity, sequence-specific binding of a trans-acting factor(s) was observed in gel shift assays. This binding was half-maximally competed by a 4.4-fold excess of nonradioactive probe and was very stable once formed (delta H [symbol: see text] dissoc. = 32 kcal/mol), suggesting that low concentrations of a high affinity binding protein(s) exist in nuclear extracts. Further support for this conclusion came from the observation that cotransfection of a plasmid containing multiple copies of the 56-bp negative element was able to relieve the negation of GME activity in a GME-56-bp-GRE reporter construct. These data directly support the role of a trans-acting factor(s) in binding to the 56-bp negative element and blocking GME activity. Collectively, these data suggest that glucocorticoid induction of TAT gene expression is subject to multiple levels of control by several new cis-acting elements and thus is much more complex than previously appreciated. PMID- 8732679 TI - Estrogen and estrogen receptor antagonists stimulate transcription from the human retinoic acid receptor-alpha 1 promoter via a novel sequence. AB - We and others previously reported that up-regulation of retinoic acid receptor alpha (RAR alpha) RNA and protein levels is elicited by estrogen in human breast cancer cells. We set out to determine the mechanism by which estrogen up regulates RAR alpha. Cloning of 500 bp of the human (h) RAR alpha 1 promoter has been reported previously; we obtained this 500-bp DNA sequence by PCR techniques from human genomic DNA and tested its activity in the context of a luciferase containing reporter vector in Hep G2 cell contransactivation assays. Estradiol elicited a 6- to 8-fold increase in luciferase activity from the reporter vector driven by hRAR alpha promoter sequence between -491 and +36 bp that was dependent on the presence of contransfected estrogen receptor (ER). Analysis of various truncated versions of this promoter sequence indicated that two regions of the sequence are sensitive to estrogen stimulation. The first resides in the region 49 to -79 bp upstream from the transcription start site and conferred approximately 2-fold activation by estrogen. This region does not contain a consensus estrogen response element, and ER binding to this DNA sequence was not observed. The second responsive sequence lies at -455 to -491 bp and conferred in additional 4- to 6-fold activation by estrogen. This upstream sequence contains two A/TGGTCA half-sites; however, direct binding of ER to this sequence was not observed. Additionally, ER DNA-binding domain mutants that are not capable of binding to DNA were just as effective as wild type ER in their ability to confer estrogen responsiveness to the RAR alpha promoter, implying that ER DNA-binding ability is not required for the estrogen-induced increase in transcriptional activity. Mutation of either half-site or of an additional immediate downstream sequence in the context of the -491 to +36 bp construct reduced the luciferase activity induction by estrogen from 6-fold to 1.5- to 2-fold. Placement of the region between -455 to -491 bp upstream of an SV40 promoter-driven luciferase vector conferred approximately 20- to 30-fold stimulation of luciferase activity by estrogen in an ER-dependent manner. The ER antagonists, 4-hydroxy-tamoxifen, keoxifene, and ICI 164384, each acted as weak agonist via the hRAR alpha promoter in contransactivation assays, exhibiting 20-30% of the efficacy that was demonstrated by estradiol. Interestingly, upon treatment of MCF7 cells with estradiol or the ER antagonists, increased levels of RAR alpha RNA and protein were observed with the antagonists as well as with estrogen. PMID- 8732681 TI - Cross-coupling between insulin and estrogen receptor in human neuroblastoma cells. AB - Insulin is a well known mitotic agent for neuroblastoma cells. Human SK-N-BE neuroblastoma cells stably transfected with the estrogen receptor, however, undergo growth arrest and differentiation when treated with insulin. These effects were shown to be due to an insulin-dependent activation of the unliganded estrogen receptor. Here, we demonstrate that this activation involves the AF-2 COOH-terminal domain of the estrogen receptor and that the communication between estrogen and insulin receptor systems occurs via selected and specific transduction signals. In fact, by the use of dominant negative and dominant positive mutants we demonstrate that p21ras is essential for insulin and estrogen receptor coupling. With pharmacological tools, we prove that PI 3'kinase does not contribute to this cross-talk and that protein kinase C triggers transduction signals that act in synergism with p21ras. These results prove the intricacy of all these intracellular paths of communication. The finding that, in neuroblastoma cells, selected signal transduction systems are involved in the insulin-dependent activation of estrogen receptor is of particular interest considering that estrogen receptor might restrict the role played by insulin during the differentiation of neural cells and interfere with its proliferative potential while allowing its regulation of other functions related to cell survival. PMID- 8732680 TI - Estrogen regulates activity of cyclin-dependent kinases and retinoblastoma protein phosphorylation in breast cancer cells. AB - Cyclin-dependent kinases (Cdk) act to regulate G1- to S-phase transition in mammalian cells. We have studied the effects of estradiol and the steroidal antiestrogen ICI 182, 780 on induction of Cdk activity and the consequent phosphorylation of retinoblastoma protein (Rb) in estrogen-responsive MCF-7 breast cancer cells. Treatment of growth-arrested MCF-7 cells with physiological concentrations of estradiol led to a time-dependent increase in Cdk2-associated and cyclin E-dependent kinase activity, which was accompanied by hyperphosphorylation of Rb and S-phase entry. Induction of both Cdk2 activity and DNA synthesis by estradiol was dose dependent and was inhibited by coadministration of ICI 182,780. Elicitation of Cdk2 activity was found to require prolonged (> 8h) estradiol exposure. Levels of cyclins E and A were unchanged in MCF-7 cells undergoing G1- to S-transit; however, synthesis and steady state levels of cyclin D1 protein were increased by estradiol. Cdk4 associated Rb kinase activity was evident in MCF-7 cells by 6 h after estradiol exposure and was inhibited by antiestrogen. Cdk2 and Cdk4 protein levels were not altered by estrogen treatment; however, faster migrating, phosphorylated Cdk2 forms increased in estradiol-treated MCF-7 cells by 12 after release from growth arrest. Cdtk-inhibitory activities, associated with p27kip-1, were eliminated from growth-arrested MCF-7 cells after treatment with estradiol but were not eliminated from cells cotreated with estradiol and ICI 182,780. These findings suggest that estradiol regulates G1 progression in MCF-7 cells through direct effects upon Cdk activation, Rb phosphorylation, and by inducing elimination of Cdk inhibitors. PMID- 8732682 TI - Characterization and cloning of STAT5 from IM-9 cells and its activation by growth hormone. AB - The interaction of GH with its receptor has been shown to lead to the phosphorylation of the signal transducer and activator of transcription (STAT) family of transcription factors. We demonstrate here that GH activates the tyrosine phosphorylation of STAT5 in the human IM-9 lymphocyte cell line. Western blotting indicates that GH also activates STAT5 in human embryonic kidney cells (293), which stably express the rabbit GH receptor. Although it has been shown previously that GH activates both STATs 1 and 3 in the 3T3-F442A mouse preadipocyte cell line, we demonstrate that GH also activates STAT5 in these cells. Using electrophoretic mobility shift assay, we examined the interaction of proteins with DNA elements containing consensus STAT-binding sequences. Proteins prepared from GH-treated 3T3-F442A cells bound to the c-sis inducible element of the human c-fos gene (m67 SIE), whereas proteins from GH-treated IM-9 cells did not. However, proteins from GH-treated IM-9 cells did interact with oligonucleotides containing either an interferon response element or the lactogenic hormone-responsive region. Treatment of IM-9 cells with interferon gamma also induced protein interactions with these elements although the complexes were distinctly different than those seen with GH treatment. Using STAT specific antibodies, we demonstrate that the GH-induced DNA-protein complex formed with the lactogenic hormone-responsive region contained STAT5, while the interferon-gamma-induced complex contained STAT1. These results implicate STAT5 as a downstream mediator of GH action in IM-9 cells. We report here the cloning of two forms of STAT5, STAT5A and STAT5B, from an IM-9 cDNA library. Northern blot analysis demonstrated multiple-forms of STAT5 mRNA in IM-9 cells. PMID- 8732685 TI - Two mutations of the lutropin/choriogonadotropin receptor that impair signal transduction also interfere with receptor-mediated endocytosis. AB - The experiments presented herein were designed to probe a potential role for the activation of the LH/CG receptor (LHR) on the receptor-mediated endocytosis of human CG (hCG). Two mutants of the rat LHR (rLHR) that bind the hormone with high affinity but are deficient in signal transduction were prepared by mutating highly conserved residues that have been previously shown to be important in signal transduction in other members of the G protein-coupled receptor family. Mutation of a highly conserved aspartic acid in the second transmembrane domain of the rLHR (designated rLHR-D383N) does not affect hCG binding but impairs signal transduction. When compared with cells expressing an equivalent density of wild type rLHR (rLHR-wt), concentration-response curves for the hCG-stimulated cAMP accumulation in cells expressing rLRH-D383N- are characterized by an 18-fold increase in the EC50 but no change in the maximal response. Cells expressing rLHR D383N also display a 4- to 5-fold increase in the half-life of internalization of hCG. Mutation of a highly conserved arginine in the second intracellular loop of the rLHR (designated rLHR-R442H) also does not affect hCG binding but impairs signal transduction. When compared with cells expressing an equivalent density of rLHR-wt, concentration-response curves for the hCG-stimulated cAMP accumulation in cells expressing rLHR-R442H are characterized by a 7-fold increase in the EC50 and a 6- to 10-fold decrease in the maximal response. Cells expressing rLHR-R442H also display a 1.5- to 2-fold increase in the half-life of internalization of hCG. These results, together with the finding that an antagonist of hCG is internalized more slowly than hCG, suggest that the activation of the LHR is needed for the efficient endocytosis of the bound hCG. PMID- 8732684 TI - Activin signaling through activin receptor type II causes the cachexia-like symptoms in inhibin-deficient mice. AB - Activins and inhibins, members of the transforming growth factor-beta superfamily, are involved in diverse physiological and developmental processes. We have previously shown that mice deficient in alpha-inhibin develop gonadal sex cord-stromal tumors at an early age. The tumor development is rapidly followed by a wasting syndrome that includes severe weight loss, hepatocellular necrosis around the central vein, and depletion of the parietal cells in the glandular stomach. The liver histology in inhibin-deficient mice is similar to the pathological effects of short-term treatment of rats and mice with recombinant activin A. Consistent with these findings, we have shown that the gonadal tumors in the inhibin-deficient mice secrete high levels of activins. In addition, Northern blot analysis has localized activin receptor type II (ActRII) to the liver. Based on these studies, we postulated that tumor-produced activins act through ActRII to cause the wasting syndrome in inhibin-deficient mice. To test this hypothesis and determine the significance of elevated levels of activin signaling through ActRII in vivo, we generated compound homozygous mutant mice deficient in both alpha-inhibin and ActRII. Despite the continued development of gonadal sex cord-stromal tumors and elevated serum levels of activin A and B, the compound homozygous mutant mice suffered no unusual weight loss, and the stomachs and livers of the majority of the mice were histologically normal. These results demonstrate that increased levels of activin signaling through ActRII in hepatocytes and the glandular stomach causes the hepatocellular necrosis and depletion of parietal cells in the glandular stomach as well as the severe weight loss in vivo. PMID- 8732683 TI - The role of the growth hormone (GH) receptor and JAK1 and JAK2 kinases in the activation of Stats 1, 3, and 5 by GH. AB - GH has been shown to activate the GH receptor (GHR)-associated tyrosine kinase JAK2 and the Src homology 2 domain-containing transcription factors Stats (signal transducers and activators of transcription) 1, 3, and 5. The present work investigates the role of GHR and JAK2 in the activation of Stats 1, 3, and 5 by GH. The ability of GH to stimulate the tyrosyl phosphorylation of these Stats was assessed in Chinese hamster ovary (CHO) cells expressing truncated and mutated GHR. GH was observed to stimulate tyrosyl phosphorylation of Stats 1, 3, and 5 in CHO cells expressing GHRs that bind JAK2 [GHR1-638 (full-length) and GHR1-454 (lacks approximately half of the cytoplasmic domain)] but not in CHO cells expressing GHR that do not bind JAK2 (GHR1-318 or GHR1-294). GH-dependent tyrosyl phosphorylation of Stat5, but not Stats 1 or 3, was reduced in CHO cells expressing GHR1-454. GH-dependent tyrosyl phosphorylation of Stats 3 and 5 was severely reduced and undetectable for Stat1 in cells expressing GHR1-454 in which tyrosines 333 and 338 (the only tyrosines phosphorylated within 1-454) are mutated to phenylalanine (GHR1-454Y333, 338F). However, GH-dependent phosphorylation of Stats 1, 3, and 5 was observed in cells expressing full-length GHR in which tyrosines 333 and 338 are mutated to phenylalanine (GHR1-638Y333, 338F) GH, whose receptor lacks previously defined Stat1- or Stat3-binding sites, was found in 3T3-F442A fibroblasts and 2fTGH-GHR cells to stimulate tyrosyl phosphorylation of JAK2 to a substantially greater extent than, and JAK1 to a similar extent as, leukemia inhibitory factor (LIF) and/or interferon gamma (IFN gamma), ligands whose receptors contains Stat3- and Stat1-binding sites and activate Stat3 and Stat1, respectively, better than GH. These findings suggest that: 1) JAK2 is required for GH-dependent phosphorylation of Stats 1, 3, and 5; 2) tyrosines 333 and/or 338 are required for maximal tyrosyl phosphorylation of Stats 1, 3, and 5; 3) Stat5 binds to a phosphorylated tyrosine(s) within amino acids 454-638 in addition to tyrosines 333 and/or 338; 4) GH stimulates tyrosyl phosphorylation of JAK1 in addition to JAK2 with JAK2 having a much greater response; 5) some Stat3 and Stat5 (and possibly Stat1) may bind to nonphosphorylated amino acids in GHR or to phosphorylated tyrosines in proteins that bind to GHR (e.g. JAK22) to be maximally activated; and 6) if JAK2, which contains Stat3-binding motifs, does serve as a docking site for some Stat proteins, Stat-JAK2 binding is likely to be more important for GH than LIF or IFN gamma in 3T3-F442A cells since GH induces 15 times more tyrosyl-phosphorylated JAK2 than LIF or IFN gamma. PMID- 8732686 TI - Calcium-sensing receptor expression and regulation by extracellular calcium in the AtT-20 pituitary cell line. AB - A 120 kDa, G protein-coupled calcium-sensing receptor (CaR) was recently identified and cloned from bovine parathyroid and rat kidney. We report here that a similar calcium-sensing receptor is also present in rat and mouse pituitary as well as in the mouse pituitary cell line, AtT-20. Fragments (383-bp) of the extracellular domain of the calcium-sensing receptor from the AtT-20 cells and mouse pituitary were amplified by RT-PCR, sequenced, and found to be identical. By Northern blot analysis, AtT-20 cells expressed a major CaR mRNA transcript of 7.5 kb and three minor transcripts of 9.5, 4.0, and 1.5 kb. Except for the 9.5-kb species, these CaR transcripts were also found to be present in mouse kidney, where the 7.5-kb transcript was again the predominant form. The presence of the CaR protein in AtT-20 cells was documented directly by fluorescence immunocytochemistry using an antibody directed against the extracellular domain of the CaR. Exposure of AtT-20 cells to increasing extracellular calcium concentrations from 0.3 t 3 mM for 24 h resulted in a 2- to 4-fold increase in the levels of CaR mRNA, but not of the RNAs for beta-actin or POMC. The CaR appeared to be functional in AtT-20 cells, since acute increases in extracellular calcium between 2 and 5 mM induced increases in the cellular content of total inositol phosphates, cytosolic calcium, and cAMP. This report suggests that pituitary cells respond to changes in extracellular calcium via a G protein coupled CaR.